PMID- 12718913 TI - Magnetofection--a highly efficient tool for antisense oligonucleotide delivery in vitro and in vivo. AB - Delivery of antisense oligodesoxynucleotides (ODN) into primary cells is a specific strategy for research with therapeutic perspectives but transfection associated difficulties. We established the technique of magnetofection to enhance ODN delivery at low toxicity and procedure time in vitro and in vivo. In vitro, target knockout was assessed at protein and mRNA levels and by measuring superoxide generation after antisense magnetofection against the p22(phox) subunit of endothelial NAD(P)H-oxidase. Under magnetic field guidance, low-dose magnetic particle-bound ODN were transfected to 84% human umbilical vein endothelial cells within 15 min followed by nuclear accumulation within 2 h, which required 24 h using standard methods. Antisense magnetofection against p22(phox) significantly decreased basal and prevented stimulated superoxide release due to loss of NAD(P)H-oxidase activity by mRNA knockout as assessed after 24 h. Knockout of endothelial phosphatase SHP-1 and connexin 37 proteins confirmed the method's efficiency. Transfection-associated toxicity was minimal. Twenty-four hours after injection of fluorescence-labeled ODN into femoral arteries of male mice, there was specific ODN uptake only into cremaster vessels exposed to magnetic fields during injection. Magnetofection is an ideal tool for delivery of functionally active ODN to difficult-to-transfect cells to study gene/protein function and a promising strategy for targeted ODN delivery in vivo. PMID- 12718914 TI - Macromolecular crystallization in the structural genomics era. PMID- 12718915 TI - Protein crystals and their growth. AB - Recent results on the associations between protein molecules in crystal lattices, crystal-solution surface energy, elastic properties, strength, and spontaneous crystal cracking are reviewed and discussed. In addition, some basic approaches to understanding the solubility of proteins are followed by an overview of crystal nucleation and growth. It is argued that variability of mixing in batch crystallization may be a source of the variation in the number of crystals ultimately appearing in the sample. The frequency at which new molecules join a crystal lattice is measured by the kinetic coefficient and is related to the observed crystal growth rate. Numerical criteria used to discriminate diffusion- and kinetic-limited growth are discussed on this basis. Finally, the creation of defects is discussed with an emphasis on the role of impurities and convection on macromolecular crystal perfection. PMID- 12718916 TI - Nucleation of protein crystals. AB - This paper introduces nucleation theory applied to crystallizing protein solutions. It is shown that the classical approach explains the available nucleation data under most conditions used for growing protein crystals for structural studies and for industrial crystallization. However, it fails to explain most experimental data on the structure of the critical clusters. It is also shown that for open systems working out of equilibrium, such as hanging-drop and counterdiffusion techniques, the geometry of the Ostwald-Myers protein solubility diagram and the number, size, and quality of the forming crystals depend not only on supersaturation but also on the rate of development of supersaturation. PMID- 12718917 TI - Macromolecular crystal growth as revealed by atomic force microscopy. AB - Direct visualization of macromolecular crystal growth using atomic force microscopy (AFM) has provided a powerful tool in the delineation of mechanisms and the kinetics of the growth process. It has further allowed us to evaluate the wide variety of impurities that are incorporated into crystals of proteins, nucleic acids, and viruses. We can, using AFM, image the defects and imperfections that afflict these crystals, the impurity layers that poison their surfaces, and the consequences of various factors on morphological development. All of these can be recorded under normal growth conditions, in native mother liquors, over time intervals ranging from minutes to days, and at the molecular level. PMID- 12718918 TI - Noninvasive in situ observation of the crystallization kinetics of biological macromolecules by confocal laser scanning microscopy. AB - High-resolution confocal laser scanning microscopy (CLSM) is a powerful tool for in situ observation and analysis of protein crystal growth kinetics. Because the resolution of CLSM is not diffraction-limited by the object, it is possible to visualize, under certain conditions, objects in molecular dimensions. A modified batch technique is applied which allows the growth kinetics of sufficiently small crystallites fixed at the lower side of a cover glass, within a hanging drop, to be studied in reflected light near the total reflection angle. A gap, or cavity, filled with solution is formed between the cover glass and the upper crystal face, which acts to fix small crystallites by hydrodynamic friction forces. The cavity height enables the propagation of molecular steps across the upper crystal face without constraint, so that the propagation velocity and geometrical parameters can be measured by CLSM. The layer growth kinetics of monoclinic crystallites of a long-acting insulin derivative (Insulin Glargine) is investigated. For a twofold supersaturation of the solution, the growth is governed by 2D nucleation at the edges of the crystallites followed by a spreading of molecular steps. The layer growth kinetics are well fitted by the simple cubic kinetic lattice model. We find that only about one of a thousand solute (protein) molecules which push a kink place due to their Brownian motion becomes really incorporated into the growing crystal. PMID- 12718919 TI - Light scattering as a diagnostic for protein crystal growth--a practical approach. AB - Static and dynamic light scattering are discussed as particularly useful tools for studying various aspects of protein crystal growth. Specific applications for prenucleation assays as well as for monitoring postnucleation growth processes are presented. Protein-protein interactions determined by light scattering, which serve as a predictor for favorable crystallization conditions as well as for protein solubility behavior, are detailed. Several precautions regarding the practical aspects of light scattering and interpretation of data are also discussed. PMID- 12718920 TI - Seeds to crystals. AB - Seeding has been critical for obtaining diffraction-quality crystals for many structures. In this article, applications and recommendations for seeding are presented based on examples from our laboratory and other groups. The implementation of seeding in high-throughput crystallization, robotics, and other emerging technologies is also discussed. PMID- 12718921 TI - New techniques in macromolecular cryocrystallography: macromolecular crystal annealing and cryogenic helium. AB - Cryocrystallography is used today for almost all X-ray diffraction data collection at synchrotron beam lines, with rotating-anode generators, and micro X ray sources. Despite the widespread use of flash-cooling to place macromolecular crystals in the cryogenic state, its use can ruin crystals, trips to the synchrotron, and sometimes even an entire project. Annealing of macromolecular crystals takes little time, requires no specialized equipment, and can save crystallographic projects that might otherwise end in failure. Annealing should be tried whenever initial flash-cooling causes an unacceptable increase in mosaicity, results in ice rings, fails to provide adequate diffraction quality, or causes a crystal to be positioned awkwardly. Overall, annealing improves the quality of data and overall success rate at synchrotron beam lines. Its use should be considered whenever problems arise with a flash-cooled crystal. Helium is a more efficient cryogen than nitrogen and will deliver lower temperatures. Experiments suggest that when crystals are cooled with He rather than N2, crystals maintain order and high-resolution data are less affected by increased radiation load. Individually or in combination, these two techniques can enhance the success of crystallographic data collection, and their use should be considered essential for high-throughput programs. PMID- 12718922 TI - The protein as a variable in protein crystallization. AB - Strategies for growing protein crystals have for many years been essentially empirical, the protein, once purified to a certain homogeneity, being mixed with a selection of crystallization agents selected in a more or less trial-and-error fashion. Screening for the correct conditions has been made easier through automation and by the introduction of commercially available crystallization kits. Many parameters can be changed in these experiments, such as temperature, pH, and ionic strength, but perhaps the most important variable has been ignored, namely the protein. The crystallization properties of a protein vary greatly: some crystallize readily, whereas others have proven extremely difficult or even impossible to obtain in a crystalline state. The possibility of altering the intrinsic characteristics of a protein for crystallization has become a feasible strategy. Some historical perspectives and advances in this area will be reviewed. PMID- 12718923 TI - RNA crystallization. AB - RNA molecules may be crystallized using variations of the methods developed for protein crystallography. As the technology has become available to synthesize and purify RNA molecules in the quantities and with the quality that is required for crystallography, the field of RNA structure has exploded. The first consideration when crystallizing an RNA is the sequence, which may be varied in a rational way to enhance crystallizability or prevent formation of alternate structures. Once a sequence has been designed, the RNA may be synthesized chemically by solid-state synthesis or it may be produced enzymatically using RNA polymerase and an appropriate DNA template. Purification of milligram quantities of RNA can be accomplished by HPLC or gel electrophoresis. As with proteins, crystallization of RNA is usually accomplished by vapor diffusion techniques. There are several considerations that are either unique to RNA crystallization or more important for RNA crystallization. Techniques for design, synthesis, purification, and crystallization of RNAs will be reviewed here. PMID- 12718924 TI - Membrane protein crystallization. AB - The need for high-resolution structure information on membrane proteins is immediate and growing. Currently, the only reliable way to get it is crystallographically. The rate-limiting step from protein to structure is crystal production. An overview of the current ideas and experimental approaches prevailing in the area of membrane protein crystallization is presented. The long established surfactant-based method has been reviewed extensively and is not examined in detail here. The focus instead is on the latest methods, all of which exploit the spontaneous self-assembling properties of lipids and detergent as vesicles (vesicle-fusion method), discoidal micelles (bicelle method), and liquid crystals or mesophases (in meso or cubic-phase method). In the belief that a knowledge of the underlying phase science is integral to understanding the molecular basis of these assorted crystallization strategies, the article begins with a brief primer on lipids, mesophases, and phase science, and the related issue of form and function as applied to lipids is addressed. The experimental challenges associated with and the solutions for procuring adequate amounts of homogeneous membrane proteins, or parts thereof, are examined. The cubic-phase method is described from the following perspectives: how it is done in practice, its general applicability and successes to date, and the nature of the mesophases integral to the process. Practical aspects of the method are examined with regard to salt, detergent, and screen solution effects; crystallization at low temperatures; tailoring the cubic phase to suit the target protein; different cubic-phase types; dealing with low-protein samples, colorless proteins, microcrystals, and radiation damage; transport within the cubic phase for drug design, cofactor retention, and phasing; using spectroscopy for quality control; harvesting crystals; and miniaturization and robotization for high-throughput screening. The section ends with a hypothesis for nucleation and growth of membrane protein crystals in meso. Thus far, the bicelle and vesicle-fusion methods have produced crystals of one membrane protein, bacteriorhodopsin. The experimental details of both methods are reviewed and their general applicability in the future is commented on. The three new methods are rationalized by analogy to crystallization in microgravity and with respect to epitaxy. A list of Web resources in the area of membrane protein crystallogenesis is included. PMID- 12718925 TI - Protein production in Escherichia coli for structural studies by X-ray crystallography. AB - The arrival of genomic sequences to the database has provided a seemingly unlimited supply of targets for protein structure determination and the possibility of solving the structure of an entire proteome. Based on our experience with the proteomes of Pyrobaculum aerophilum and Mycobacterium tuberculosis, we have developed a simple strategy for the production of proteins for structural studies by X-ray crystallography. Our scheme demonstrates a strong protein target commitment and includes the expression of genes from these organisms in Escherichia coli. These proteins are expressed with affinity tags and purified for characterization and crystallization. We have identified protein solubility and crystallization as the two major bottlenecks in the process toward the determination of protein structures by X-ray diffraction. Strategies to overcome these bottlenecks are discussed. PMID- 12718926 TI - Membrane protein structural biology: the high throughput challenge. AB - Membrane proteins represent roughly one-third of the proteins encoded in the genome, yet fewer than 1% of the proteins are of known structure. High-throughput crystallography offers the hope of correcting this imbalance. In order for large scale membrane protein structural biology to realize its full promise, however, significant technical challenges must be overcome, the two most substantial being facile protein overexpression and reliable methods for crystal growth. PMID- 12718927 TI - High-throughput protein crystallization. AB - High-throughput structural biology is a focus of a number of academic and pharmaceutical laboratories around the world. The use of X-ray crystallography in these efforts is critically dependent on high-throughput protein crystallization. The application of current protocols yields crystal leads for approximately 30% of the input proteins and well-diffracting crystals for a smaller fraction. Increasing the success rate will require a multidisciplinary approach that must invoke techniques from molecular biology, protein biochemistry, biophysics, artificial intelligence, and automation. PMID- 12718928 TI - Maximum-likelihood crystallization. AB - The crystallization facility of the TB Structural Genomics Consortium, one of nine NIH-sponsored structural genomics pilot projects, employs a combinatorial random sampling technique in high-throughput crystallization screening. Although data are still sparse and a comprehensive analysis cannot be performed at this stage, preliminary results appear to validate the random-screening concept. A discussion of statistical crystallization data analysis aims to draw attention to the need for comprehensive and valid sampling protocols. In view of limited overlap in techniques and sampling parameters between the publicly funded high throughput crystallography initiatives, exchange of information should be encouraged, aiming to effectively integrate data mining efforts into a comprehensive predictive framework for protein crystallization. PMID- 12718929 TI - A deliberate approach to screening for initial crystallization conditions of biological macromolecules. AB - A method to rationally predict crystallization conditions for a previously uncrystallized macromolecule has not yet been developed. One way around this problem is to determine initial crystallization conditions by casting a wide net, surveying a large number of chemical and physical conditions to locate crystallization leads. A facility that executes the rapid survey of crystallization lead conditions is described in detail. Results and guidelines for the initial screening of crystallization conditions, applicable to both manual and robotic setups, are discussed. PMID- 12718930 TI - The promise of macromolecular crystallization in microfluidic chips. AB - Microfluidics, or lab-on-a-chip technology, is proving to be a powerful, rapid, and efficient approach to a wide variety of bioanalytical and microscale biopreparative needs. The low materials consumption, combined with the potential for packing a large number of experiments in a few cubic centimeters, makes it an attractive technique for both initial screening and subsequent optimization of macromolecular crystallization conditions. Screening operations, which require a macromolecule solution with a standard set of premixed solutions, are relatively straightforward and have been successfully demonstrated in a microfluidics platform. Optimization methods, in which crystallization solutions are independently formulated from a range of stock solutions, are considerably more complex and have yet to be demonstrated. To be competitive with either approach, a microfluidics system must offer ease of operation, be able to maintain a sealed environment over several weeks to months, and give ready access for the observation and harvesting of crystals as they are grown. PMID- 12718931 TI - Efficient protein crystallization. AB - High-throughput molecular biology and crystallography advances have placed an increasing demand on crystallization, the one remaining bottleneck in macromolecular crystallography. This paper describes three experimental approaches, an incomplete factorial crystallization screen, a high-throughput nanoliter crystallization system, and the use of a neural net to predict crystallization conditions via a small sample (approximately 0.1%) of screening results. The use of these technologies has the potential to reduce time and sample requirements. Initial experimental results indicate that the incomplete factorial design detects initial crystallization conditions not previously discovered using commercial screens. This may be due to the ability of the incomplete factorial screen to sample a broader portion of "crystallization space," using a multidimensional set of components, concentrations, and physical conditions. The incomplete factorial screen is complemented by a neural network program used to model crystallization. This capability is used to help predict new crystallization conditions. An automated, nanoliter crystallization system, with a throughput of up to 400 conditions/h in 40-nl droplets (total volume), accommodates microbatch or traditional "sitting-drop" vapor diffusion experiments. The goal of this research is to develop a fully-automated high throughput crystallization system that integrates incomplete factorial screen and neural net capabilities. PMID- 12718932 TI - A fully integrated protein crystallization platform for small-molecule drug discovery. AB - Structure-based drug discovery in the pharmaceutical industry benefits from cost efficient methodologies that quickly assess the feasibility of specific, often refractory, protein targets to form well-diffracting crystals. By tightly coupling construct and purification diversity with nanovolume crystallization, the Structural Biology Group at Syrrx has developed such a platform to support its small-molecule drug-discovery program. During the past 18 months of operation at Syrrx, the Structural Biology Group has executed several million crystallization and imaging trials on over 400 unique drug-discovery targets. Here, key components of the platform, as well as an analysis of some experimental results that allowed for platform optimization, will be described. PMID- 12718933 TI - Protein crystallization by capillary counterdiffusion for applied crystallographic structure determination. AB - Counterdiffusion crystallization in capillary is a very simple, cost-effective, and practical procedure for obtaining protein crystals suitable for X-ray data analysis. Its principles have been derived using well-known concepts coupling the ideas of precipitation and diffusion mass transport in a restricted geometry. The counterdiffusion process has been used to simultaneously screen for optimal conditions for protein crystal growth, incorporate strong anomalous scattering atoms, and mix in cryogenic solutions in a single capillary tube. The crystals obtained in the capillary have been used in situ for X-ray analysis. The implementation of this technique linked to the advancement of current crystallography software leads to a powerful structure determination method consolidating crystal growth, X-ray data collection, and ab initio phase determination into one without crystal manipulation. We review the historical progress of counterdiffusion crystallization, its application to X-ray crystallography, and ongoing tool development for high-throughput protein structure determination. PMID- 12718934 TI - Therapeutic immune response induced by electrofusion of dendritic and tumor cells. AB - To elicit a therapeutic antitumor immune response, dendritic cells (DCs) have been employed as a cellular adjuvant. Among various DC-based approaches, fusion of DCs and tumor cells potentially confers not only DC functionality, but also a continuous source of unaltered tumor antigens. We have recently demonstrated successful generation of fusion hybrids by a large-scale electrofusion technique. The immunogenicity and therapeutic potential of fusion hybrids were further analyzed in a model system of a murine melanoma cell line expressing beta galactosidase (beta-gal) as a surrogate tumor antigen. A single vaccination with fusion hybrids plus IL-12 induced a therapeutic immune response against 3-day established pulmonary metastases. This immunotherapy was beta-gal specific and involved both CD4 and CD8 T cells. In vitro, fusion hybrids stimulated specific IFN-gamma secretion from both CD4 and CD8 immune T cells. They also nonspecifically induced IL-10 secretion from CD4 but not CD8 T cells. Compared to other DC loadings, our results demonstrate the superior immunogenicity of fusion. The current technique of electrofusion is adequately developed for clinical use in cancer immunotherapy. PMID- 12718935 TI - Florid pulmonary inflammatory responses in mice vaccinated with Antigen-85 pulsed dendritic cells and challenged by aerosol with Mycobacterium tuberculosis. AB - Mice immunized by the intranasal route with dendritic cells harvested from the lungs and then pulsed with Ag85 (LDC-Ag85) were able to prime naive CD4(+) T cells in vivo. As a result splenic CD4(+) T cells from these immunized mice were able to produce IFNgamma following culture with Mycobacterium tuberculosis infected antigen presenting cells. Hematoxylin and eosin stained lung sections from LDC-Ag85 immunized mice after they had been exposed to aerosol challenge with M. tuberculosis showed a florid infiltration of macrophages and lymphocytes into granulomas and parenchymal tissues when compared to lung sections from control groups implanted with dendritic cells pulsed with ovalbumin. In addition, using immunohistochemistry, these tissues appeared to have more CD4(+) and CD8(+) cells than the control groups. This was confirmed by flow cytometric analysis which showed that lung cell digests contained increased numbers of CD4 and CD8 interferongamma secreting cells. Despite this increase however, no evidence was seen that indicated that the LDC-Ag85 immunized mice were more resistant to M. tuberculosis infection than mice immunized with LDC pulsed with an irrelevant protein. Instead, the potent inflammatory response in the LDC-Ag85 resulted in serious consolidation of the lung tissue. PMID- 12718936 TI - Induction of CD69 activation molecule on human neutrophils by GM-CSF, IFN-gamma, and IFN-alpha. AB - The CD69 glycoprotein is an early activation antigen of T and B lymphocytes but it expression is induced in vitro on cells of most hematopoietic lineages, including neutrophils after stimulation with PMA or fMLP. In this study, we investigated whether CD69 expression on human neutrophils could be modulated by inflammatory or anti-inflammatory cytokines (IL-1beta, IL-2, IL-4, IL-6, IL-8, IL 10, IL-12, IL-18, G-CSF, GM-CSF, TNF-alpha, TGF-beta, IFN-alpha, IFN-gamma). Resting neutrophils from healthy subjects did not express CD69 on the cell surface; moreover, a preformed intracellular pool of CD69 was not evident in these cells. CD69 was barely detectable on these cells after overnight incubation in medium while overnight incubation with GM-CSF, IFN-gamma or IFN-alpha significantly induced CD69 expression on neutrophils with GM-CSF appearing to be the most potent inducer. This induction was dependent on a new protein synthesis as it was significantly inhibited by cycloheximide (about 50% inhibition). CD69 cross-linking on GM-CSF-primed neutrophils sinergized with LPS and increased TNF alpha production and secretion suggesting a role for CD69-positive neutrophils in the pathogenesis and maintenance of different inflammatory diseases. PMID- 12718937 TI - CD38 ligation plays a direct role in the induction of IL-1beta, IL-6, and IL-10 secretion in resting human monocytes. AB - CD38 signaling, either induced by ligation with specific agonistic monoclonal antibody (mAb) or after interaction with CD31, its cognate counter-receptor, is involved in release of IL-1beta, IL-6, and IL-10 cytokines in resting human monocytes. CD38 ligation by the F(ab')(2) IB4 mAb did not induce signals relevant for cytokine secretion and the block of the Fcgamma receptor I (FcgammaRI) by anti-CD64 or FcgammaRII by anti-CD32 mAb did not inhibit CD38-mediated IL-1beta release. Dimerization or multimerization of the CD38 molecule by: (i) cross linking of the receptor ligated by F(ab')(2) or by (ii) increasing CD38 expression by treating monocytes with IFNgamma were able to restore the truncated CD38-mediated signals involved in cytokine secretion. These data indicate that CD38 receptor-mediated signals operate directly suggesting a Fcgamma receptorial surface molecule independent activation pathway. The key element for the receptor mediated signaling is represented by surface density of CD38 on resting monocytes. PMID- 12718938 TI - IL-7-mediated protection of pro and pre-B cells from the adverse effects of corticosterone. AB - The studies herein demonstrate that Interleukin-7 (IL-7) promotes survival of murine pro- and pre-B cells against stress levels of corticosterone (Cs). In short-term, 16-h, bone marrow cultures IL-7 abrogated Cs-induced apoptosis and cell cycle arrest in pro-B cells by decreasing apoptosis 60% and completely restoring the cell cycle. IL-7 also reduced Cs-induced apoptosis by 36% in pre-B cells and 24% in IgM(+) B cells, but did not restore deficits in the cell cycle. Among pro- and pre- B cells, substantial protection against high, pharmacological, levels of Cs was also provided by IL-7. Interestingly, stem cell factor, while reducing spontaneous apoptosis in pro-B cells, did not protect against Cs-induced death, either alone or with IL-7. In conclusion, IL-7 has potential immunotherapeutic value since it provides substantial protection to pro and pre-B cells against the adverse effects of Cs. PMID- 12718939 TI - Age-related changes in mature CD4+ T cells: cell cycle analysis. AB - T cell proliferative responses decrease with age, but the mechanisms responsible are unknown. We examined the impact of age on memory and naive CD4(+) T cell entry and progression through the cell cycle using acridine orange to identify cell cycle stage. For both subsets, fewer stimulated cells from old donors were able to enter and progress through the first cell cycle, with an increased number of cells arrested in G(0) and fewer cells in post G(0) phases. The number of dead cells as assessed by sub-G(0) DNA was also significantly greater in the old group. CD4(+) T cells from old mice also exhibited a significant reduction in clonal history as assessed by CFSE staining. This was associated with a significant decline in cyclin D2 mRNA and protein. We propose that decreases in cyclin D2 are at least partially responsible for the proliferative decline found in aged CD4(+) T cells. PMID- 12718940 TI - Relationships between burn size, immunosuppression, and macrophage hyperactivity in a murine model of thermal injury. AB - Burn injury induces immune dysfunction and alters numerous physiological parameters. While clinical studies indicate that burn injury size profoundly impacts patient immune status, only limited experimental studies have systematically addressed its impact on immune functional parameters. In the present study, mice were subjected to burn injuries of varying sizes and splenic immune cells (splenocytes and macrophages) were isolated 7 days thereafter. Burn injury suppressed splenic T-cell proliferation in an injury size-dependent manner that correlated with the release of the immunosuppressive mediators PGE(2) and nitric oxide. In addition, a shift towards an immunosuppressive Th-2 cytokine profile and a hyperactive macrophage phenotype (increased release of inflammatory mediators) was observed post-injury, however, this effect was in part independent of burn size. Thus, unlike patient survival data, burn injury-induced changes in immune function do not necessarily correlate with the size of the injury. PMID- 12718942 TI - Excitement vs burnout. PMID- 12718941 TI - Antagonistic effect of picolinic acid and interferon-gamma on macrophage inflammatory protein-1alpha/beta production. AB - The L-tryptophan catabolite, picolinic acid (PA), is an activator of macrophage effector functions and an inducer of macrophage inflammatory protein-1alpha (MIP 1alpha) and -1beta (MIPs). We have investigated the regulation of PA-induced MIPs production in mouse macrophages. We demonstrated a dose- and time-dependent downregulation of MIPs mRNA by the Th1 cytokine, IFN-gamma, that was associated with inhibition of intracellular chemokine production and secretion. This effect was IFN-gamma-specific because MIPs induction was unaffected by the Th2 cytokines, IL-10 and IL-4, or the proinflammatory stimulus, LPS. Moreover, MIPs downregulation by IFN-gamma was dependent on both mRNA destabilization and gene transcription inhibition. These results demonstrate that MIP-1alpha/beta production by macrophages is a tightly regulated process resulting from the interaction between inhibitory stimuli derived from the immune system and stimulatory signals of non-immunologic origin. The antagonistic effect of PA and IFN-gamma on MIPs production may be important for the regulation of the inflammatory responses in vivo. PMID- 12718943 TI - Delayed diagnosis of right-sided diaphragmatic rupture. AB - Diaphragmatic rupture is uncommon and results from either blunt or penetrating trauma. Right-sided traumatic diaphragmatic ruptures are easily missed. We present a case of rupture of the right diaphragm, which highlights the difficulty of confirming the correct diagnosis. PMID- 12718944 TI - The lived experience of violence in accident and emergency. AB - AIM: To investigate the lived experience of violence of nurses in one A&E department. RATIONALE: Following an extensive literature review, it was elicited that there was very little evidence of the psychological impact of violence in A&E. BACKGROUND/INTRODUCTION: Violence in the health service has increased over the last decade associated with a corresponding increase in the published literature on this topic. While violence in A&E departments would appear common, little evidence exists on the psychosocial impact of these incidents on A&E nurses. RESEARCH METHODS: A qualitative domain using a phenomenological philosophy was utilised. Sampling was of a volunteer nature and data were collected using interview. Nurses were sampled from one major acute hospital in Northern Ireland. RESULTS: Participants expressed feelings of frustration, anger, and fear. Support from colleagues was greatly valued and lack of feedback from management was cited as being common. DISCUSSION: Frustration and anger were synthesised into the "why me" principle with nurses not knowing why they were the focus for attack. Violent incidents were associated with a sense of isolation but colleague support developed a sense of belonging. CONCLUSIONS: Whilst a useful insight was provided, more research is essential into this emotive topic. PMID- 12718945 TI - A literature review into pain assessment at triage in accident and emergency departments. PMID- 12718946 TI - Case review: ejection from a motor vehicle. PMID- 12718947 TI - Being a non-urgent patient in an emergency care unit--a strive to maintain personal integrity. AB - The aim of this study was to analyse and describe experiences of being a non urgent patient in an ECU (emergency care unit). Eleven non-urgent patients were interviewed. The research approach was inductive and interpretative. Seven tentative interpretations and an interpreted whole, i.e., an existential interpretation, revealed that the informants tried to be 'good' patients by not demanding much attention from nursing personnel, in an attempt to maintain good relations with the nurses in order to be assured of a positive reception. As health related problems jeopardize personal integrity, patients cannot afford the risk of being looked upon as inappropriate clients in the ECU. PMID- 12718948 TI - The holistic nursing care of patients with minor injuries attending the A&E department. AB - Philosophy, theoretical frameworks, and concepts of nursing are applied to the particular context of accident and emergency nursing. From this the research question evolves; the researcher seeks to explore the accident and emergency nurses' perceptions of the concept of holistic nursing care, a concept that is included within their department's philosophy for nursing. An ethnographic approach is applied to the research to gather rich descriptive data through exploratory interviews. The data were collected and analysed simultaneously, categories of data were developed, and these are presented here in the findings. The categories were scrutinised for patterns, similarities, and dissimilarities between the data; recurrent themes emerge. These themes shed light upon the meanings of holism in care to the respondents and offer an understanding of their cultural systems that affect the provision of holistic nursing care, these are discussed and analysed within the contexts of the existing knowledge base. The study identifies issues that adversely affect the provision of holistic nursing care, notably fragmentation of care, dissemination of nursing information, and privacy. The implications for practice are discussed, and recommendations include action research for practical issues and further research into the provision of holistic nursing care incorporating the perceptions of patients who receive care in this setting. PMID- 12718949 TI - Managing thrombolysis in the accident and emergency department. AB - The optimal management of acute myocardial infarction (AMI) today is founded upon the 'open artery' theory and driven by the need for early reperfusion strategies. Numerous randomised controlled trials demonstrate the unequivocal benefit of thrombolytic therapy in reducing mortality and improving long-term prognosis. These benefits are most striking when the thrombolytic therapy is given early. The National Service Framework for Coronary Heart Disease introduced national standards to reduce both 'call to needle' and 'door to needle' times to further augment early treatment. Subsequently many hospitals established 'fast track', 'nurse-led' or 'nurse initiated' thrombolysis within both the Accident and Emergency and Coronary Care Departments. This paper reviews these strategies and considers the benefits and limitations associated with the role of the 'thrombolysis nurse'. Thrombolysis nurses work to agreed protocols and play a fundamental role in the assessment and appropriate management of patients with acute MI. The evidence suggests that thrombolysis nurses are safe and effective in their practice and make a significant contribution to patient outcomes and enhance interprofessional education and practice. PMID- 12718950 TI - Making the link--an impact evaluation of one Dublin hospital's accident & emergency department's liaison nurse service. AB - The Accident & Emergency (A&E) liaison nurse service (LNS) was introduced at the Mater Misericordiae Hospital, Dublin as a quality improvement initiative in June 1996. It aimed to improve and enhance communication channels between the hospital and general practitioners (GPs) in the hospital's catchment area. A quantitative study was conducted to evaluate the A&E LNS, in partial fulfilment of a Master's degree in Nursing Science at the University of Dublin, Trinity College, Dublin. The primary users (90 GPs and 52 A&E staff, n=142) of the service were asked to complete a questionnaire comprising open and closed questions. Descriptive analysis using parametric and non-parametric tests was carried out, using the Statistical Package for the Social Sciences (SPSS version 9), on the collected data. Content analysis was used to analyse the open-ended questions. The findings of the study suggest that the A&E LNS positively affects communication between the hospital and GPs, and favourably affects the continuity and quality of patient care. The study respondents strongly supported the provision of an A&E LNS and expressed positive attitudes towards the quality of the service provided. This piece of research provides a basic understanding of the role, function and scope of practice of the A&E liaison nurse specialist. Implications for practice and areas for improvement are also outlined and identified. PMID- 12718951 TI - Professional portfolios: all you need to know and were afraid to ask. PMID- 12718955 TI - Standard forms--helpful clinical and investigative tools for determining cause and timing of inflicted brain damage in children. PMID- 12718956 TI - Use of the Child Abuse Potential inventory as a measure of treatment outcome. PMID- 12718957 TI - Dynamic prediction characteristics of the Child Abuse Potential Inventory. AB - OBJECTIVES: Risk measures are commonly used to evaluate outcomes in child abuse prevention and intervention programs. This study examined whether pre intervention to post-intervention changes on the Child Abuse Potential Inventory (CAP) Abuse Scale corresponded to actual changes in risk for future reports of maltreatment and evaluated the validity of several algorithms for classifying clinically significant change. METHOD: Participants in the study were 459 parents participating in any one of 27 community-based family preservation and family support programs. Most parents were low-income mothers with a variety of social risk indicators, about a third of whom would be classified as high-risk by the CAP Abuse Scale. Participants were administered the CAP at program enrollment, then at completion of the intervention (median time=150 days), then followed for an average of approximately 2 years for future official maltreatment reports. Dynamic predictive validity of the CAP Abuse Scale was modeled by comparing survival models using a time-dependent structure of pre- and post-intervention scores to identically structured models using only a pre-intervention score. RESULTS: Pre-intervention CAP Abuse Scale scores demonstrated incremental future predictive validity. However, score changes failed to correspond to changes in likelihood of future abuse. Models using pre-intervention scores only were more predictive than time-dependent score models, and pre-intervention scores were better predictors than post-intervention scores of post-intervention CPS referrals. Common algorithms for classifying clinically significant change yielded results that could be counter-intuitive and misleading. For example, participants classified as improved on these algorithms were actually at similar or even higher risk than those classified as unchanged or worse. CONCLUSIONS: The results strongly supported the static predictive validity of the CAP and the use of the CAP for screening purposes. The results did not support the dynamic predictive validity of the CAP. Results of exploratory analyses suggested the possibility that the changes observed on the CAP Abuse Scale reflected changes in subscales assessing subjective distress or parenting attitudes, which may be markers for initial risk but when changed, do not necessarily translate into actual changes in future maltreatment behavior. Although replication and extension are needed before drawing firm conclusions, the current study raises questions about the common practice of using risk instruments as proxy measures for child maltreatment risk in intervention and prevention programs. PMID- 12718958 TI - Teachers' beliefs about the effects of child abuse. AB - OBJECTIVE: Teachers' expectations about the effects of physical and emotional abuse on children's classroom behaviors were examined in this study. Not only do teachers have to decide if a particular child is the victim of abuse, they may also have to contend with changes in that child's classroom behavior. METHOD: Teachers generated what they thought were typical outcomes of physical and emotional abuse on children's classroom behavior. RESULTS: Responses generally fell into the following categories: lowered self-esteem, heightened aggression, academic difficulties, and poor social interaction skills. Teachers who mentioned lowered self-esteem were more likely to generate it as the result of emotional abuse rather than physical abuse. CONCLUSIONS: Teachers' expectations generally mirrored research findings as to the actual effects of child abuse. Suggestions are made to incorporate the results in training programs designed to increase teachers' self-confidence in reporting potential cases of child abuse. PMID- 12718959 TI - [Current understanding about intergenerational transmission of child sexual abuse]. AB - OBJECTIVE: The aim of this article is to review what is currently understood about intergenerational transmission of child sexual abuse (CSA). METHOD: CSA transmission is discussed first from the point of view of men CSA survivors who become sexually abusive, and then from the perspective of mothers who survived CSA whose children have been sexually abused. Mechanisms that may help us understand how CSA is transmitted from one generation to another are described. More specifically, focus is given to those mechanisms that might differentiate CSA survivors who break the cycle of abuse from those who perpetuate it. RESULTS: In light of the research reviewed, it seems that the transmission of CSA is far from inevitable, since one-third of sexually abusive men and half of sexually abused children's mothers mentioned having been sexually abused in their childhood. Because of the retrospective method used in many studies, causal links could not be established. However, some mechanisms have been proposed in order to better understand the phenomenon of CSA. Severity of abuse, attachment relationships with parental figures, as well as dissociative symptoms that follow the abuse were identified. Dissociative symptomatology appeared to be a determining factor in understanding the cycle of CSA. CONCLUSIONS: More studies on CSA transmission are needed to understand the mechanisms that are involved in that cycle, as well as to develop effective strategies to treat and prevent CSA. PMID- 12718960 TI - Children with reported histories of sexual abuse: utilizing multiple perspectives to understand clinical and psychosocial profiles. AB - OBJECTIVE: The current study examines multiple empirically based perspectives (i.e., child, caregiver, and clinician) of behavior and functioning as they contribute to the clinical and psychosocial profile of children (aged 5 to 17.5 years) with reported histories of sexual abuse. METHOD: A large, multi-site data set of children referred into Comprehensive Community Mental Health Services both with and without reported histories of sexual abuse, was examined. Seven hundred and fifty-nine children with a reported history of sexual abuse were compared to 2722 without such a history on caregiver and child reported behavior, clinician rated functioning, diagnosis, demographic variables, and life challenges. RESULTS: The multiple perspectives contributed unique and specific information to regression models: caregiver-reported behavior contributed information about externalizing behavior while child-reported behavior added information about internalizing behavior and clinician ratings about self-harmful behavior. Children with reported histories of sexual abuse were also more likely to be female, Caucasian, and have reported histories of life challenges (e.g., physical abuse, substance use, running away). Child sexual abuse was associated with higher rates of depression and anxiety diagnoses, and lower rates of substance abuse, conduct, and attention deficit disorder diagnoses. CONCLUSIONS: The findings indicate that the profile of children entering into Comprehensive Community Mental Health Services with reported histories of sexual abuse, as compared to those without such histories, is complex and best understood via multiple perspectives. Caregiver, child and clinician rated information, when taken together, provide a comprehensive clinical and psychosocial profile around which to plan and implement individualized service plans. PMID- 12718961 TI - Why children tell: a model of children's disclosure of sexual abuse. AB - OBJECTIVE: The present study investigated variables associated with delay of disclosure of child sexual abuse and tested a model of time to disclosure. METHOD: Data were obtained for 218 alleged child sexual abuse victims whose cases had been referred to District Attorneys' Offices. Five variables were posited to influence the delay between an abusive event and children's disclosure of that event to a reporting adult: child's age, gender, type of abuse experienced (intrafamilial or extrafamilial), perceived responsibility for the abuse, and fear of negative consequences of disclosure. These variables were used to create a model of factors influencing children's disclosure of sexual abuse. RESULTS: Results indicated that age, type of abuse, fear of negative consequences, and perceived responsibility all contributed to predicting time to disclosure. There was significant support for the model, suggesting that children who were older, came from incestuous families, felt greater responsibility for the abuse, and feared negative consequences of disclosure took longer to disclose. CONCLUSIONS: Children's cognitive appraisal of others' tolerance of disclosure of child sexual abuse, and their own perceptions of responsibility for the abuse, are crucial to the decision to disclose. When evaluating children for possible sexual abuse, developmental, cognitive, and socio-emotional factors need to be taken into consideration. PMID- 12718962 TI - Empathy and narcissism in a sample of child abuse perpetrators and a comparison sample of foster parents. AB - OBJECTIVE: The purpose of this research was to study the personality variables of empathy and narcissism in a sample of child abuse perpetrators and a comparison sample of foster parents, conceptualized as nonabusive parents, in order to gain further understanding of perpetrators and to provide clues for intervention. METHOD: The sample consisted of two groups: physically and emotionally abusive parents (n=52) and foster parents (n=101). Participants responded to three instruments: the Interpersonal Reactivity Index (IRI), an instrument measuring individual differences in empathy, and two instruments measuring narcissism: the Narcissistic Personality Inventory (NPI) and the Hypersensitivity Narcissism Scale (HSNS). RESULTS: Statistically significant differences were found between the two groups on three of the four subscales of the IRI: perspective-taking, empathic concern, and personal distress. Based on the definition of these subscales, the abusive parents as compared to the foster parents were not able to take the perspective of another or see things from another's viewpoint, showed less warmth, compassion and concern for others, and experienced difficulty in tense interpersonal situations. Statistically significant differences were found for the two groups on three of the six subscales of the NPI: authority, exhibitionism, and superiority, and on the HSNS. The abusive parents demonstrated less self-confidence, a greater lack of impulse control and were more narcissistic than their foster parent counterparts. CONCLUSIONS: The results suggest that it is how the perpetrators experience aversive behavior in their children that may provoke them to physically and emotionally abuse their children. Their self-centeredness in addition to their deficiencies in empathy may cause them to experience their children's misbehavior as an affront to their authority. Implications for treatment are made from this conceptualization of parental abuse. PMID- 12718963 TI - Men who kill their own children: paternal filicide incidents in contemporary Fiji. AB - OBJECTIVE: Filicide, or parental murder of offspring, constitutes a major portion of lethal violence perpetrated against children worldwide. Despite the global nature of the phenomenon, researchers have focused their studies on the developed industrialized societies with the consequent neglect of small, developing societies. Second, there is a paucity of empirical data on child homicide committed by fathers. This study therefore explores the nature and extent of paternal filicides in Fiji, a non-Western society, and the social and cultural forces underlying them in order to enhance our knowledge of the phenomenon. METHOD: Information was obtained from a number of sources, including (a) a police homicide logbook, (b) newspaper reports of homicide, and (c) detailed interviews conducted with criminal justice and medical personnel. Information from these data sources were consolidated to construct case histories of paternal filicides. These cases were then analyzed for dominant themes. Case illustrations are presented in the text. RESULTS: Several of the study's findings are congruent with other studies of paternal filicides: poor, working class fathers were the offenders in all cases. As a corollary, their victims were from low socioeconomic backgrounds. Regarding location, paternal filicides occurred in the home of the offender and victim. The filicides were the culmination of stresses and strains associated with marital disharmony and excessive corporal child-control strategies. CONCLUSIONS: The general conclusion of this study is that further research in non-Western societies has the potential to increase our understanding of the social factors and processes involved in paternal child murders. We will then be better positioned to craft effective intervention strategies. PMID- 12718966 TI - Gas-phase mechanism for dioxin formation. AB - We consider the formation of dioxin in the gas phase through the combination of phenoxy radicals in the context of a recently published scenario regarding the possibility of forming highly chlorinated benzenes in poorly mixed systems. It is demonstrated that the production of free chlorine in fuel lean combustion systems and subsequent mixing and quenching with fuel rich mixtures leads to chlorine atoms as the main reactive radicals. Under such conditions high concentrations of chlorinated phenoxy radicals can be formed. This leads to the formation of concentrations of dioxins that are consistent with recent measurements of Sidhu et al. [Combust. Flame 100 (1995) 11]. It suggests that there may be a gas-phase channel for the dioxin formation in addition to the usually assumed surface catalyzed process. PMID- 12718967 TI - Chlorination of dibenzofuran and dibenzo-p-dioxin vapor by copper (II) chloride. AB - Dibenzofuran (DF) is formed from phenol and benzene in combustion gas exhaust streams prior to particle collection equipment. Subsequent chlorination at lower temperatures on particle surfaces is a potential source of chlorinated dibenzofuran (CDF). Gas streams containing 8% O(2) and approximately 0.1% DF vapor were passed through particle beds containing copper (II) chloride (0.5% Cu, mass) at temperatures ranging from 200 to 400 degrees C to investigate the potential for CDF formation during particle collection. Experiment duration was sufficient to provide an excess amount of DF (DF/Cu=3). The efficiency of DF chlorination by CuCl(2) and the distribution of CDF products were measured, with effects of temperature, gas velocity, and experiment duration assessed. Results of a more limited investigation of dibenzo-p-dioxin (DD) chlorination by CuCl(2) to form chlorinated DD (CDD) products are also presented. The efficiency of DF/DD chlorination by CuCl(2) was high, both in terms of CuCl(2) utilization and DF/DD conversion. Total yields of Cl on CDF/CDD products of up to 0.5 mole Cl per mole CuCl(2) were observed between 200 and 300 degrees C; this suggests that nearly 100% CuCl(2) was utilized, assuming a conversion of two moles of CuCl(2) to CuCl per mole Cl added to DD/DF. In a short duration experiment (DF/Cu=0.3), nearly 100% DF adsorption and conversion to CDF was achieved. The degree of CDF chlorination was strongly dependent on gas velocity. At high gas velocity, corresponding to a gas-particle contact time of 0.3 s, mono-CDF (MCDF) yield was largest, with yields decreasing with increasing CDF chlorination. At low gas velocity, corresponding to a gas-particle contact time of 5 s, octa-CDF yield was largest. DF/DD chlorination was strongly favored at lateral sites, with the predominant CDF/CDD isomers within each homologue group those containing Cl substituents at only the 2,3,7,8 positions. At the higher temperatures and lower gas velocities studied, however, broader isomer distributions, particularly of the less CDD/CDF products, were observed, likely due to preferential destruction of the 2,3,7,8 congeners. PMID- 12718968 TI - Effects of copper chloride on formation of polychlorinated dibenzo-p-dioxins in model waste incineration. AB - Combustion experiments in a laboratory-scale fluidized-bed reactor were conducted to elucidate the effects of copper chloride as a catalyst on polychlorinated dibenzo-p-dioxins (PCDDs) formation in municipal waste incineration. We used model wastes with and without copper chloride (CuCl(2) x 2H(2)O), both of which contained polyvinyl chloride as a chlorine source. Combustion temperature was set to 900 degrees C, and the amount of air supplied was twice the stoichiometric ratio. The experimental setup was carefully planned to suppress the influences of experimental conditions except the waste composition. Results of these experiments showed that copper chloride in the waste increased the amount of PCDDs formed and made the homologue profile to shift towards the highly chlorinated species. Copper chloride contributes to the PCDDs formation by promoting chlorination, whereby the reaction is important in that organic matter is chlorinated directly by copper compounds. Copper chloride did not exert a great influence on the isomer distribution patterns of PCDDs, while there appeared a significant difference in the case of PCDFs. This points out the difference of the major formation mechanisms between PCDDs and PCDFs. PCDDs are less formed by the catalytic reactions from carbon/polycyclic aromatic hydrocarbons than PCDFs in our experimental conditions. PMID- 12718969 TI - Modeling aerosol formation in opposed-flow diffusion flames. AB - The microstructures of atmospheric pressure, counter-flow, sooting, flat, laminar ethylene diffusion flames have been studied numerically by using a new kinetic model developed for hydrocarbon oxidation and pyrolysis. Modeling results are in reasonable agreement with experimental data in terms of concentration profiles of stable species and gas-phase aromatic compounds. Modeling results are used to analyze the controlling steps of aromatic formation and soot growth in counter flow configurations. The formation of high molecular mass aromatics in diffusion controlled conditions is restricted to a narrow area close to the flame front where these species reach a molecular weight of about 1000 u. Depending on the flame configuration, soot formation is controlled by the coagulation of nanoparticles or by the addition of PAH to soot nuclei. PMID- 12718970 TI - Development of a measuring technique for simultaneous in situ detection of nanoscaled particle size distributions and gas temperatures. AB - Point measurements of time-resolved LII signals have been performed in sooting premixed low pressure flames. Soot particle size distribution and gas temperature in these flames are known from independent measurements. This data is used to validate parameters of an improved LII model, where special emphasis is taken on the accurate modelling of mass and heat transfer rates. Using this model particle size distributions and gas temperatures can be estimated from time-resolved LII signals using non-linear regression. Standard numerical methods are applied. An experimental setup is presented, which allows measuring one-dimensional maps of particle size distribution and gas temperature. The technique is based on the one dimensional and time-resolved detection of LII signals using a Streak camera. PMID- 12718971 TI - Comparative analysis of the structure of carbon materials relevant in combustion. AB - The determination of the structure of carbon materials is an analytical problem that join the research scientific communities involved in the chemical characterization of heavy fuel-derived products (heavy fuel oils, coal-derived fuels, shale oil, etc.) and of carbon materials (polycyclic aromatic compounds, tar, soot) produced in many combustion processes. The knowledge of the structure of these "difficult" fuels and of the carbon materials produced by incomplete combustion is relevant to research for the best low-environmental impact operation of combustion systems; but an array of many analytical and spectroscopic tools are necessary, and often not sufficient, to attempt the characterization of such complex products and in particular to determine the distribution of molecular masses. In this paper the size exclusion chromatography using N-methyl-pyrrolidinone as eluent has been applied for the characterization of different carbon materials starting from typical carbon species, commercially available like polyacenaphthylene, carbon black, naphthalene pitch up to combustion products like soot and soot extract collected in fuel-rich combustion systems. Two main fractions were detected, separated and molecular weights (MWs) determined by comparison with polystyrene standards: a first fraction consisted of particles with very large molecular masses (>100000 u); a second fraction consisted of species in a relatively small MW range (200-600 u). The distribution of these fractions changes in dependence on the carbon sample characteristics. Fluorescence spectroscopy applied on the fractions separated by size-exclusion chromatography has been used and comparatively interpreted giving indications on the differences and similarities in chemical structure of such different materials. PMID- 12718972 TI - Spectroscopic behavior of oxygenated combustion by-products. AB - The oxygenated species, massively produced in the energy production plants based on combustion processes, constitute one of the most numerous categories of hazardous air pollutants. Therefore, development of real time diagnostic tools are needed in order to study their formation during combustion processes and to reveal their presence both in the exhaust and in the atmosphere. In this work, oxygenated compounds were identified inside fuel-rich premixed ethylene/air flames by means of ultraviolet fluorescence spectroscopy with the support of qualitative chemical analysis of the sampled combustion gases. Strong band progression, typical of aldehydic functionality, were recognized in fluorescence spectra (lambda(exc)=355 nm) measured in the early oxidation region of premixed flames varying the equivalence ratio from 3.0 up 21.6. Downstream of the oxidation region, spectroscopic signatures of pyrolytic species were found to prevail on those peculiar of oxygenated compound. The position and the extension of the two main flame zones were found to depend on the flame conditions (C/O ratio) due to the effect of the C/O ratio on the temperature history along the flame axis. This correlation was interpreted on the basis of the measured axial temperature profiles. PMID- 12718973 TI - Detection of combustion formed nanoparticles. AB - UV-visible extinction and scattering and two extra situ sampling techniques: atomic force microscopy (AFM) and differential mobility analysis (DMA) are used to follow the evolution of the particles formed in flames. These particle sizing techniques were chosen because of their sensitivity to detect inception particles, which have diameters, d<5 nm, too small to be observed with typical particle measurement instrumentation. The size of the particles determined by AFM and DMA compares well with the size determined by in situ optical measurements, indicating that the interpretation of the UV-visible optical signal is quite good, and strongly showing the presence of d=2-4 nm particles. UV-visible extinction measurements are also used to determine the concentration of d=2-4 nm particles at the exhausts of practical combustion systems. A numerical model, able to reproduce the experimentally observed low coagulation rate of nanoparticles with respect to soot particles, is used to investigate the operating conditions in the combustion chamber and exhaust system for which 2-4 nm particles survive the exhaust or grow to larger sizes. Combustion generated nanoparticles are suspected to affect human and environmental health because of their affinity for water, small size, low rate of coagulation, and large surface area/weight ratio. The ability to isolate nanoparticles from soot particles in hydrosols collected from combustion may be useful for future analysis by a variety of techniques and toxicological assays. PMID- 12718974 TI - Structure of inorganic and carbonaceous particles emitted from heavy oil combustion. AB - The combustion of heavy fuel oil for power generation is a great source of carbonaceous and inorganic particle emissions, even though the combustion technologies and their efficiency are improving. The information about the size distribution function of the particles originated by trace metals present into the fuels is not adequate. In this paper, we focused our attention on the larger distribution mode of both the carbonaceous and metallic particles. Isokinetic sampling was performed at the exhausts of two typical heavy oil flames and the samples were size-segregated by mean of an 8-stages Andersen impactor. Further investigation performed on the samples using electronic microscopy coupled with X ray analysis (EDX) evidenced the presence of solid spherical particles, called plerosphere(1) as analogy with cenosphere, with typical dimensions ranging between 200 nm and 2-3 microm, whose atomic composition contains a large amount of the trace metals present in the parent oils (Fe, V, Ni, etc). EDX analyses revealed that the metal concentration increases as the plerosphere dimension decreases. PMID- 12718975 TI - Monitoring of fuel consumption and aromatics formation in a kerosene spray flame as characterized by fluorescence spectroscopy. AB - The large presence of aromatic compounds in distillate fossil fuels should allow, in line of principle, to follow the fuel consumption and/or the presence of unburned fuel in a high temperature environment like a burner or the exhaust of combustion systems by exploiting the high fluorescence emission of aromatic fuel components. To this aim an UV-excited fluorescence source has to be used since the aromatic fuel components are strongly fluorescing in the UV region of the emission spectrum. In this work UV-excited laser induced fluorescence (LIF) diagnostics was applied to spray flames of kerosene in order to follow the fuel consumption and the formation of aromatic species. A strong UV signal was detected in the spray region of the flame that presented a shape similar to that found in the LIF spectra preliminary measured on the cold spray and in the room temperature fluorescence of fuel solutions. The decrease of UV signal along the spray flame region was associated to the consumption of the fuel, but more difficult seems to be the attribution of a broad visible emission, that is present downstream of the flame. The visible emission feature could be assigned to flame-formed PAH species contained in the high molecular weight species, hypothesizing that their fluorescence spectra are shifted toward the visible for effect of the high temperature flame environment. PMID- 12718976 TI - Sampling and analysis of ambient dioxins in northern Taiwan. AB - In this study, ambient air samples were taken concurrently in the vicinity area of a large-scale municipal waste incinerator (MWI) and the background area for measuring polychlorinated dibenzo-p-dioxins and furans (PCDD/Fs) concentrations from November 1999 through July 2000 in northern Taiwan. The results obtained from eighteen ambient air samples indicate that the mean PCDD/F concentration of seventeen 2,3,7,8-substituted congeners in wintertime (188-348 fg-I-TEQ/m(3)) is significantly higher than that measured in summertime (56-166 fg-I-TEQ/m(3)). In addition, the seasonal PCDD/F concentrations are compared with the ambient air quality data including CO, NO(2), PM(10) and TSP sampled from Taipei area to gain better insights. It indicates that the variation of ambient air PCDD/F concentrations is closely correlated with that of PM(10) concentrations. Besides, the results indicate that the I-TEQ concentration of ambient air in sampling site B (directly downwind of the MWI) is of the highest while the sampling site A (upwind of MWI) is of the lowest among all sampling sites. This implies that existing MWI can be a significant emitter of PCDD/Fs in this area. Furthermore, the patterns of the PCDD/F congener distribution at all sampling sites (including the background site in Taoyuan) are quite similar. OCDD concentration is of the highest among seventeen PCDD/F congeners investigated and accounts for about 35% of the total concentration. As for the I-TEQ concentrations, 2,3,4,7,8-PeCDF is the most significant contributor, generally being responsible for 30-45% of the total I-TEQ values depending on the sampling sites and seasons. PMID- 12718977 TI - Reaction of arsenic vapor species with fly ash compounds: kinetics and speciation of the reaction with calcium silicates. AB - In coal combustion systems, the partitioning of arsenic between the vapor and solid phases is determined by the interaction of arsenic vapors with fly ash compounds under post-combustion conditions. This partitioning is affected by gas solid reactions between the calcium components of the ash particles and arsenic vapors. In this study, bench scale experiments were conducted with calcium compounds typical of coal-derived fly ash to determine product formation, the extent of reaction and reaction rates when contacted by arsenic oxide vapors. Experiments conducted with arsenic trioxide (As(4)O(6(g))) vapors in contact with calcium oxide, di-calcium silicate and mono-calcium silicate over the temperature range 600-1000 degrees C indicated that these solids were capable of reacting with arsenic vapor species in both air and nitrogen. Calcium arsenate was the observed reaction product in all the samples analyzed. Maximum capture of arsenic occurred at 1000 degrees C with calcium oxide being the most effective of the three solids over the range of temperatures studied. Using a shrinking core model for a first order reaction and the results from intrinsic kinetic experiments conducted in air, the reaction rate constants were found to be 1.4 x 10(-3)exp( 2776/T) m/s for calcium oxide particles, 7.2 x 10(-3)exp(-3367/T) m/s for di calcium silicate particles and 5.5 x 10(-3)exp(-3607/T) m/s for mono-calcium silicate particles. These results therefore suggest that any calcium present in fly ash can react with arsenic vapor and capture the metal in water-insoluble forms of the less hazardous As(V) oxidation state. PMID- 12718978 TI - Impact of the composition of combustion generated fine particles on epithelial cell toxicity: influences of metals on metabolism. AB - Inhaled airborne particulate matter (PM) represents a potentially significant health hazard to humans. Exposure to PM strongly correlates with pulmonary inflammation and incidences of severe respiratory distress, including increased hospital admissions for breathing disorders, asthma, emphysema, and chronic bronchitis. PM generated from the combustion of fuel oils and coals contain a number of water-soluble transition metals including Fe, V, and Zn. We have evaluated the impact of PM types with varying composition collected from the combustion of oils and coals on the health and metabolism of lung cell cultures. Three colorimetric assays (sulforhodamine B (SRB), Janus green, and MTT) have been adapted to quantify the impact of PM on rat lung alveolar type II epithelial cells (RLE-6TN cells). The PM toxicity metrics evaluated were inhibition of cell proliferation (SRB and Janus green) and inhibition of cellular metabolism (MTT). Cell proliferation is inhibited in a consistent dose-dependent manner by PM concentrations from 25 to 250 microg/ml. At a level of 100 microg/ml, oil-derived PM diminishes cell metabolism by as much as 40% relative to controls; the degree of inhibition is strongly dependent on PM particle size and metal content. Conversely, coal-derived PM at the same dosage diminishes cell metabolism by no more than 20% relative to controls. All three assays provide highly repeatable results and consistent toxicity rankings of the PMs evaluated. Overall, metabolic inhibition as measured by the MTT assay was deemed the most appropriate metric for PM toxicity, primarily due to its applicability with in vivo-like confluent cell monolayers. PMID- 12718979 TI - Inhalation health effects of fine particles from the co-combustion of coal and refuse derived fuel. AB - This paper is concerned with health effects from the inhalation of particulate matter (PM) emitted from the combustion of coal, and from the co-combustion of refuse derived fuel (RDF) and pulverized coal mixtures, under both normal and low NO(x) conditions. Specific issues focus on whether the addition of RDF to coal has an effect on PM toxicity, and whether the application of staged combustion (for low NO(x)) may also be a factor in this regard. Ash particles were sampled and collected from a pilot scale combustion unit and then re-suspended and diluted to concentrations of approximately 1000 microg/m(3). These particles were inhaled by mice, which were held in a nose-only exposure configuration. Exposure tests were for 1 h per day, and involved three sets (eight mice per set) of mice. These three sets were exposed over 8, 16, and 24 consecutive days, respectively. Pathological lung damage was measured in terms of increases in lung permeability. Results show that the re-suspended coal/RDF ash appeared to cause very different effects on lung permeability than did coal ash alone. In addition, it was also shown that a "snapshot" of lung properties after a fixed number of daily 1-h exposures, can be misleading, since apparent repair mechanisms cause lung properties to change over a period of time. For the coal/RDF, the greatest lung damage (in terms of lung permeability increase) occurred at the short exposure period of 8 days, and thereafter appeared to be gradually repaired. Ash from staged (low NO(x)) combustion of coal/RDF appeared to cause greater lung injury than that from unstaged (high NO(x)) coal/RDF combustion, although the temporal behavior and (apparent) repair processes in each case were similar. In contrast to this, coal ash alone showed a slight decrease of lung permeability after 1 and 3 days, and this disappeared after 12 days. These observations are interpreted in the light of mechanisms proposed in the literature. The results all suggest that the composition of particles actually inhaled is important in determining lung injury. Particle size segregated leachability measurements showed that water soluble sulfur, zinc, and vanadium, but not iron, were present in the coal/RDF ash particles, which caused lung permeabilities to increase. However, the differences in health effects between unstaged and staged coal/RDF combustion could not be attributed to variations in pH values of the leachate. PMID- 12718980 TI - Genetic susceptibility to nickel-induced acute lung injury. AB - Human exposure to insoluble and soluble nickel compounds is extensive. Besides wide usage in many industries, nickel compounds are contained in cigarette smoke and, in low levels, in ambient particulate matter. Soluble nickel particulate, especially nickel sulfate (NiSO(4)), has been associated with acute lung injury. To begin identifying genes controlling susceptibility to NiSO(4), mean survival times (MSTs) of eight inbred mouse strains were determined after aerosol exposure. Whereas A/J (A) mice were sensitive, C57BL/6J (B6) mice survived nearly twice as long (resistant). Their offspring were similarly resistant, demonstrating heritability as a dominant trait. Quantitative trait locus (QTL) analysis of backcross mice generated from these strains identified a region on chromosome 6 significantly linked to survival time. Regions on chromosomes 1 and 12 were suggestive of linkage and regions on chromosomes 8, 9, and 16 contributed to the response. Haplotype analysis demonstrated that QTLs on chromosomes 6, 9, 12, and 16 could explain the MST difference between the parental strains. To complement QTL analysis results, cDNA microarray analysis was assessed following NiSO(4) exposure of A and B6 mice. Significant expression changes were identified in one or both strains for >100 known genes. Closer evaluation of these changes revealed a temporal pattern of increased cell proliferation, extracellular matrix repair, hypoxia, and oxidative stress, followed by diminished surfactant proteins. Certain expressed sequence tags clustered with known genes, suggesting possible co-regulation and novel roles in pulmonary injury. Together, results from QTL and microarray analyses of nickel-induced acute lung injury survival allowed us to generate a short list of candidate genes. PMID- 12718981 TI - [Extreme solutions in human reproduction]. PMID- 12718982 TI - [Insulin resistance and polycystic ovary syndrome]. AB - Polycystic ovary syndrome (PCOS) is a frequent disease, characterized by disturbed ovarian function with hyperandrogenism. Anovulation is secondary to an absence of follicular dominance. Apart from a primary ovarian defect, insulin resistance is observed in PCOS women, even in the absence of overweight. This insulin resistance could be secondary to a defect in the insulin transduction pathway, mainly by a defect in receptor phosphorylation. It enhances hyperandrogenism as it increases ovarian androgen production. Therefore treating insulin resistance by weight loss or drugs reducing insulin resistance might improve fertility of PCOS women. Metformin has been shown to reduce ovarian production, enhance ovulatory cycles and in some cases increase fertility. However, there are few randomized studies on large numbers of patients to prove an effect on pregnancies as well as on the occurrence of early pregnancy loss. There are currently no recommendation on dose and duration of metformin treatment. It is noteworthy that metformin has no authorization in France to be prescribed apart from diabetic patients' care. Considering the medical care of PCOS women, the cardiovascular risk needs to be taken into account. Therefore hypertension, dyslipidemia and diabetes must be treated in those women who need to be followed carefully all over their life. PMID- 12718983 TI - [Prospective clinical and sonographic assessment of uterine artery embolization as the treatment of symptomatic uterine leiomyomata]. AB - OBJECTIVE: To evaluate the effectiveness of the uterine artery embolization as the treatment of symptomatic uterine leiomyomata. PATIENTS AND METHODS: Eighty five women with symptoms caused by uterine leiomyomata underwent uterine artery embolization as an alternative to surgery from january 1997 to june 2000. The effectiveness of this method was evaluated by clinical and sonographic examination. RESULTS: The recession average was of 18.9 months. There were ten failures. We had immediate failures (n = 5) with a case of technical failure, one endometrium cancer, one adenomyosis, one larger subserosal leiomyomata and one parametrial leiomyomata. We had recurrences (n = 5) with the occurrence of new leiomyomatas (1 intramural and 3 submucosal) and an evolution of previous leiomyomata. The average volume reduction was 51% for the uterus and 65% for the main fibroid at one year follow-up. Minor complications occurred in 5%. Permanent amenorrhoea was observed for 3.75% of the women. Using cox model, no predictive factors of embolisation effectiveness were found. DISCUSSION AND CONCLUSION: In the treatment of symptomatic uterine leiomyoma, uterine artery embolization is an effective alternative to surgery. After one year and half, we had 12.5% of failures. PMID- 12718984 TI - [Transfer of obstetric patients in a pregnant and postpartum condition to an intensive care unit: eighty-two cases in Burkina Faso]. AB - OBJECTIVE: To review all obstetric admissions to an intensive care unit (ICU) of an African hospital. PATIENTS AND METHODS: Retrospective analysis of the records of all obstetric patients admitted to the ICU of Souro Sanou Hospital in Burkina Faso, from January 1st, 1996, to June 30, 1998. RESULTS: Eighty-two patients out of 6119 deliveries were transferred to the ICU, which meant a 1.34% transfer rate. These transfers concerned young patients (mean age of 24 years), coming originally from outlying maternities in 52.4% and having already given birth in 64.63% of the cases. The two main diagnoses at the ICU were: eclampsia and septic shock. The large majority of the patients (73 out of 82) had at least one bad prognosis factor at admission at the ICU. A maternal mortality rate of 60% was noted, the main risk factors for mortality being acute respiratory condition and severe anemia. CONCLUSION: Mortality of obstetric patients admitted to ICUs is very high in our setting. Establishing an ICU within the obstetric unit or early detection of cases to be transferred (scoring system?) should improve the prognosis. PMID- 12718985 TI - [Systematic hysteroscopy prior to in vitro fertilization]. AB - OBJECTIVE: To evaluate the benefits of a diagnostic hysteroscopy prior to in vitro fertilization. PATIENTS AND METHODS: We retrospectively studied 145 patients who underwent ICSI during a period of 6 months. Office hysteroscopy was systematically performed before the first stimulation cycle. If pathological findings were revealed, appropriate medical or surgical treatment was given. RESULTS: Pathological patterns were observed in 45% of hysteroscopies. Endometritis, polyps and myomas and mucosal diseases were the most frequently observed. The patients aged over 38 years didn't show higher rate of pathology (29% vs 27% for the younger patients). The treatment of pathologies gave the same pregnancy rate than the normal cavities. Patients with endometritis were treated with antibiotics and 40% of them became pregnant in the following cycle. DISCUSSION AND CONCLUSION: Systematic hysteroscopy prior to IVF-ICSI showed to be an effective investigation that could improve the pregnancy rate. PMID- 12718986 TI - [Comparison of two methods of cryoconservation of sperm when in very small numbers]. AB - OBJECTIVE: In case of severe oligozoospermia or following testicular biopsy in case of azoospermia, fertilization remains possible by intracytoplasmic sperm injection (ICSI). However, if there are only a few spermatozoa available to be preserved for a delayed ICSI, these gametes must be cryopreserved according to a specific method. We have found that, using the straw classical technique, less than 2 sperm were recovered at thawing when 20 sperm were frozen and no sperm were available following freezing-thawing of only 1 to 10 sperm per straw. Then, the feasibility of programmed ICSI is always uncertain in these cases and unnecessary medical acts (ovarian stimulation and egg collection) as well as patient disappointment are frequent. MATERIAL AND METHODS: We have tested a new technique by introducing (with an ICSI pipette) 1 to 100 sperm in a microdrop (0.5 microl) of freezing medium setting in a culture dish under paraffin oil. RESULTS: Following freezing-thawing the dish, all (100%) of the frozen spermatozoa were recovered. DISCUSSION AND CONCLUSION: Since only motile sperm were frozen, the sperm which are non motile after thawing (more than 50%) are nevertheless considered usable for ICSI. PMID- 12718987 TI - [Cystic lymphangiomas of the mesentery and the epiploon. About two cases]. AB - Two cases of cystic lymphangiomas of the mesentery and the epiploon are hereby related. The authors underline the following points: first, the rarity of these tumors along with the absence of significant clinical features makes the differential diagnosis with ovary tumors a difficult task. Second, although abdominal ultra sonography, completed by CT scan, allows the hypothesis of lymphangioma, only histological examination will confirm the diagnosis. There is no other treatment but complete exeresis, with a good prognosis and exceptional recurrences. PMID- 12718988 TI - [Surrogate mothers and ovarian transposition: two attitudes to be considered in young women with cervical carcinoma. A case report]. AB - In cases of cervical cancer, there are 2 major advantages to preserving the ovaries, with or without transposition: hormone function is maintained during subsequent cancer treatment and patient quality of life is improved. We report the first case of pregnancy in a surrogate mother following stimulation of a transposed ovary before irradiation and chemotherapy for a squamous cell carcinoma of the uterine cervix. Because of the wide dissemination of information on the technical progress in this area, patients are now in a position to make therapeutic choices that are no longer guided by strictly medical considerations. PMID- 12718989 TI - [Menopause, andropause and sexual function]. AB - After about fifty years of age, at the time of the menopause and the andropause (androgenic deficiency), men as well as women can have the same type of complaint, but it is facing time that women have a different view from men. Time is coded into a woman's body, whereas it is by 'social' time that men are more often determined. On the sexual level there is a slow-down in the sexual reactions of both men and women. One should not forget, however, that the cure for sexual problems in women sometimes involves the treatment of male sexual dysfunction (erection problems, premature ejaculation, absence of ejaculation). Therapeutically, hormonal replacement therapy is undoubtedly a help, if there are no medical contra-indications and if the patient is in agreement. If the complaint is also sexual then a more specific, therapeutic treatment may often be necessary, associating hormone treatment with erection inducers. PMID- 12718990 TI - [Surgery for uterine sarcoma: review of the literature and recommendations for the standard surgical procedure]. AB - Hysterectomy and bilateral salpingo-oophorectomy is the gold standard for the surgical procedure of uterine sarcoma in case of tumor limited to the uterine corpus. Omentectomy and lymphadenectomy should be performed in carcinosarcoma. In others histologic subtypes, lymphadenectomy should be performed only in patients with enlarged nodes discovered at the time of the surgical procedure. Surgical resection should be ideally performed without uterine parcelling out and so using a laparotomy in order to avoid this risk. PMID- 12718991 TI - [Biological monitoring of autoimmune diseases during pregnancy]. AB - This review focuses on auto-immune diseases which frequently affect women and can have interactions with pregnancy: lupus erythematosus (LES), antiphospholipide syndrome (SAPL), Sjogren's syndrome (GS), rheumatoid arthritis (PR) and auto immune thyroiditis. LES may flare at the end of a pregnancy and during post partum. Its biological monitoring during pregnancy is well established. SAPL is at risk of sterility, prematurity, Hellp syndrome, eclampsia and retroplacental hematoma. The main risk, actually risk is foetal loss by placental ischemia, which has to be well known as 2 randomised studies have proven the efficacy of treatments with aspirin +/- subcutaneous heparin. LES, GS and PR can both be associated with anti SS-A +/- anti SS-B antibodies linked to a risk of congenital auriculo-ventricular block, which is fortunately low (less than 5% of the cases). Auto-immune thyroiditis are often revealed during pregnancy and may be enhanced during the six first months of post-partum. PMID- 12718992 TI - [Difficult embryo transfer: what can be done in practice?]. AB - Embryo transfer is an important step in the success of IVF treatment. One must avoid, at all cost, difficult transfers provoking bleeding, uterine contractions, and the retention of the embryo in the cervix, or even, its expulsion. Evaluation before IVF treatment, including a trial transfer, should allow one to evaluate the transfer step, anticipate problems, and thus improve treatment. The difficult insertion into the womb is most often associated with the presence of curves, which can be easily overcome by using pre-curved catheters. PMID- 12718993 TI - [Prevention of port-site metastasis after laparoscopy]. PMID- 12718994 TI - [Prevention of port-site metastasis after laparoscopy]. PMID- 12718995 TI - [Carcinologic results and fertility after conservative treatment of invasive epithelial ovarian cancer]. PMID- 12718996 TI - [Carcinologic results and fertility after conservative treatment of invasive epithelial ovarian cancer]. PMID- 12718997 TI - [The P.M.S.I. Enlargement: Evaluation System of Perinatal Care Networks in Bourgogne, France]. PMID- 12718998 TI - [There are reasons to continue hormone replacement therapy longer than 10 years]. PMID- 12718999 TI - [Hormone replacement therapy in menopause: continue or stop the prescription after 10 years?]. PMID- 12719000 TI - [Hormone replacement therapy in menopause: continue or stop the prescription after 10 years? We wish to give a epidemiological explanation]. PMID- 12719001 TI - [Clinical practice recommendations. Risk of premature birth with intact membranes. Conclusions (2002)]. PMID- 12719002 TI - Approaching a new era for hepatitis C virus therapy: inhibitors of the NS3-4A serine protease and the NS5B RNA-dependent RNA polymerase. AB - The treatment of chronic disease caused by the hepatitis C virus (HCV) is an unmet clinical need, since current therapy is only partially effective and limited by undesirable side effects. The viral serine protease and the RNA dependent RNA polymerase are the best-studied targets for the development of novel therapeutic agents. These enzymes have been extensively characterized at the biochemical and structural level and thus used to set up screening assays for the identification of selective inhibitors. These efforts lead to the discovery of several classes of compounds with potential antiviral activity. The hepatitis C virus does not replicate in the laboratory. The formidable challenge posed by the difficulty of developing cell-based assays and preclinical animal systems has been partially overcome with several alternative approaches. The development of new assays permitted the optimization of enzyme inhibitors leading eventually to molecules with the desired drug-like properties, the most advanced of which are being considered for clinical trials. PMID- 12719003 TI - Enterocin CRL35 inhibits late stages of HSV-1 and HSV-2 replication in vitro. AB - The replication of herpes simplex virus (HSV) type 1 and 2 in Vero cells is inhibited in the presence of enterocin CRL35 (ECRL), a bacteriocin produced by Enterococcus faecium CRL35. Attempts to resolve the mode of action of ECRL indicate that virus adsorption and penetration are not affected. Instead, a late step of virus multiplication is hindered since the addition of 100 microg/ml of ECRL at 8h post infection still causes a 90% inhibition of virus release. The effect of ECRL on HSV antigen expression was studied by immunofluorescence using a polyclonal serum and a monoclonal antibody against glycoprotein D (gamma protein). These studies indicated that ECRL impeded the second round of infection, apparently as a consequence of the inhibition of glycoprotein D expression. The replication of syncytial mutants of HSV-1 was significantly inhibited at a ECRL concentration of 25 microg/ml. Both the percentage of fused cells and the polykaryocyte size were affected. Studies on the effect of ECRL on viral protein synthesis showed that in the presence of ECRL, HSV late gamma proteins were not synthesized. From these findings, it is concluded that inhibition of HSV spreading by ECRL is due to the prevention of mainly late glycoprotein synthesis. PMID- 12719004 TI - Comparative in vitro effects of AZT and extracts of Ocimum gratissimum, Ficus polita, Clausena anisata, Alchornea cordifolia, and Elaeophorbia drupifera against HIV-1 and HIV-2 infections. AB - The effects of Ocimum gratissimum (GHX-2), Ficus polita (GHX-6), Clausena anisata (GHX-7), Alchornea cordifolia (GHX-26), Elaeophorbia drupifera (GHX-27), and AZT on in vitro HIV-1 and HIV-2 replication and cytopathicity were compared. All plant extracts inhibited HIV-1 strain HTLVIII(B) cytopathicity, the leaves of GHX 2 and the seeds of GHX-26 having high antiviral indices (110 and 90, respectively). Against HIV-2 strain GH1, the EC(50) values ranged from <0.005 to 0.075 mg/ml when treatment was started at 40min after virus adsorption, except for GHX-7 which showed only moderate activity and GHX-26 which had no activity. When treatment was delayed for 2h, the plant extracts, unlike AZT, were still very effective against HIV-2. Likewise, only the plant extracts were able to attain EC(90) values when high multiplicity of infection (MOI) with HIV-1 strain GH3 was used when treatment was delayed for 2h. In Molt-4 cocultures with Molt 4/HIV, early cytopathic effect (CPE) of cell fusion was unaffected by AZT but was completely inhibited by all plants at noncytotoxic concentrations. In addition, GHX-27 was selectively toxic to Molt-4/HIV cells. The plant extracts also inhibited HIV-1 reverse transcriptase (RT) activity at EC(50) values of <0.01 0.03 mg/ml. HIV-1 proviral DNA copying as determined in a polymerase chain reaction, was completely inhibited by GHX-2 and GHX-6 at 0.011 and 0.015 mg/ml, respectively. GHX-26 and GHX-27 showed only very moderate activity. PMID- 12719005 TI - Inhibition of HSV-1 replication and reactivation by the mutation-insensitive transcription inhibitor tetra-O-glycyl-nordihydroguaiaretic acid. AB - Methylated derivatives of nordihydroguaiaretic acid (NDGA)were previously shown to be potent mutation-resistant inhibitors of herpes simplex virus type 1 (HSV-1) which target Sp1 protein binding to critical viral promoters. The hydrophobic nature of these agents, however, renders them relatively water-insoluble and, therefore, limits their applicability. We report here on the anti-HSV-1 properties of a related but water-soluble glycylated derivative of NDGA, tetra-O glycyl-NDGA (G(4)N). In yield reduction assays, G(4)N inhibited replication of laboratory and clinical strains of wild type HSV-1 and ACV-resistant (HSV-1(R)) strains of HSV-1 in a dose-dependent manner, with average IC(50) values of 4.7 and 3.2 microM against wild-type and HSV-1(R) strains, respectively. An MTT-based cytotoxicity assay revealed a TC(50) value of 73.2 microM for G(4)N on Vero cells, with no reduction in viability detected at concentrations below 30 microM. Similar to its methylated counterparts, G(4)N was found to inhibit transcription of the HSV-1 ICP4 gene, a major immediate early viral regulator, and gel mobility shift assays showed it can block Sp1 protein binding to cognate sites on the ICP4 promoter. In anticipation of its potential use as a systemic anti-HSV-1 agent, we tested G(4)N in a murine trigeminal ganglia (TG) explant model system, and found G(4)N was able to prevent HSV-1 reactivation from explanted and cultured latently infected TG. PMID- 12719006 TI - Cyanovirin-N binds to the viral surface glycoprotein, GP1,2 and inhibits infectivity of Ebola virus. AB - Ebola virus (Ebo) causes severe hemorrhagic fever and high mortality in humans. There are currently no effective therapies. Here, we have explored potential anti Ebo activity of the human immunodeficiency virus (HIV)-inactivating protein cyanovirin-N (CV-N). CV-N is known to potently inhibit the infectivity of a broad spectrum of HIV strains at the level of viral entry. This involves CV-N binding to N-linked high-mannose oligossacharides on the viral glycoprotein gp120. The Ebola envelope contains somewhat similar oligosaccharide constituents, suggesting possible susceptibility to inhibition by CV-N. Our initial results revealed that CV-N had both in vitro and in vivo antiviral activity against the Zaire strain of the Ebola virus (Ebo-Z). Addition of CV-N to the cell culture medium at the time of Ebo-Z infection inhibited the development of viral cytopathic effects (CPEs). CV-N also delayed the death of Ebo-Z-infected mice, both when given as a series of daily subcutaneous injections and when the virus was incubated ex vivo together with CV-N before inoculation into the mice. Furthermore, similar to earlier results with HIV gp120, CV-N bound with considerable affinity to the Ebola surface envelope glycoprotein, GP(1,2). Competition experiments with free oligosaccharides were consistent with the view that carbohydrate-mediated CV N/GP(1,2) interactions involve oligosaccharides residing on the Ebola viral envelope. Overall, these studies broaden the range of viruses known to be inhibited by CV-N, and further implicate carbohydrate moieties on viral surface proteins as common viral molecular targets for this novel protein. PMID- 12719007 TI - Novel antiviral agent tetraglycylated nordihydroguaiaretic acid hydrochloride as a host-dependent viral inhibitor. AB - A water soluble derivative of nordihydroguaiaretic acid (NDGA), G(4)N (2), synthesized by reaction of NDGA (1) with N,N-dimethylglycine in the presence of dicyclohexylcarbodiimide and dimethylaminopyridine and then with HCl(g) (Scheme 1), competes effectively with the DNA binding domain of recombinant Sp1 protein for binding to the human immunodeficiency virus (HIV) LTR as demonstrated by an electrophoretic mobility-shift assay (EMSA). By blocking Sp1 binding to the HIV LTR, G(4)N suppresses Sp1-regulated HIV Tat transactivation and replication in cultured cells with an IC(50) of 12 microM similar to that of 3'-O-methyl-NDGA as we have previously reported. In addition simian immunodeficiency virus (SIV) replication was completely inhibited by G(4)N at 5.0 microM. G(4)N showed no toxic effect to 174 x CEM cells and H9 cells at 100 microM. PMID- 12719008 TI - Repression of HPV E6-activated RSV promoter activity by anti-cancer agents. AB - Human papillomavirus E6 forms a complex with p53 tumor suppressor and E6 associated protein, leading to the degradation of p53 via the ubiquitination pathway, resulting in the oncogenesis of cervical carcinomas. Several viral and cellular gene promoters were shown to be transactivated by E6 oncogene. In this study, to understand the role of transcription activity of E6 related to cervical carcinogenesis, the effect of cervical cancer drugs on E6 induced transcription activity has been elucidated. Several viral promoter (RSV, CMV, SV40, and HIV) luciferase reporter gene constructs, and eukaryotic E6 expression vector were prepared as an E6 transcription analysis system and an exogenous E6 protein source, respectively. It was shown that the promoters of RSV, SV40, and HIV, but not CMV, were transactivated by HPV 16 E6 in cervical cancer cell line. Several known cervical cancer drugs were investigated for their effects on transcription activity of E6 in SiHa stably transfected with E6-responsive promoters. Cervical cancer drugs consistently reduced luciferase activity, in transfectants with RSV luc (SiHa/pRSV-luc, KCTC 0427BP) E6 mRNA also. Thus, in this study, we have demonstrated that the promoters of RSV, HIV, and SV40 were transactivated by E6 in cervical cancer cells. Three cervical cancer drugs downregulated RSV-luc transcription and E6 expression by a p53 independent pathway. RSV-luc promoter analysis system could be useful for understanding the role of transcription activity of E6 related to cervical cancer and also for screening drugs against cervical cancers caused by HPV infection. PMID- 12719009 TI - Interferon, ribavirin, 6-azauridine and glycyrrhizin: antiviral compounds active against pathogenic flaviviruses. AB - Ribavirin, interferon-alpha (IFN-alpha), 6-azauridine and glycyrrhizin were tested in vitro for their antiviral activities against 11 pathogenic flaviviruses belonging to principal antigenic complexes or individual serogroups of medical importance: dengue, Japanese encephalitis, mammalian tick-borne and yellow fever virus (YFV) groups. Antiviral activity was estimated by the reduction of the cytopathic effect of each flavivirus in Vero cells and by the reduction in virus titer. Cytotoxicity was evaluated by determining the inhibition of Trypan blue exclusion in confluent cell cultures and by the evaluation of the inhibitory effect on cell growth. The specificity of action of each tested compound was estimated by the selectivity index (CC(50)/EC(50)). IFN-alpha proved to be a selective and potent inhibitor of the replication of the 11 tested pathogenic flaviviruses. Ribavirin and 6-azauridine proved to be active on the replication of the 11 tested pathogenic flaviviruses at the concentrations which did not alter normal cell morphology, but they were not selective inhibitors when selectivity indices were evaluated with regard to the inhibition of cell growth because of their cytostatic effect. Glycyrrhizin inhibited the replication of flaviviruses at high non-cytotoxic concentrations. These antiflavivirus compounds should be further evaluated for their efficacy in the treatment of flavivirus infections in vivo. PMID- 12719010 TI - Semi-quantitative detection of viral RNA in influenza A virus-infected mice for evaluation of antiviral compounds. AB - The aim of the study was to establish a murine model for sensitive screening of potential compounds with in vitro anti-influenza A virus activity. The evaluation in this in vivo model is based on semi-quantitative detection of viral RNA using one-step reverse transcriptase polymerase chain reaction (RT-PCR). After intranasal infection of fully-conscious mice with influenza A virus, the viral load of the respiratory tract tissues was investigated. Peaks were observed in the nasopharynx between Days 1 and 4, in the trachea on Day 4, and in the lungs between Days 4 and 7 post infection. The elimination of virus correlated with the appearance of specific serum antibodies. After 4 days of treatment with zanamivir, trachea and lungs revealed negative RT-PCR results, whereas viral load in the nasopharynx was significantly reduced. In conclusion, the virus spread in the described murine model is similar to upper respiratory tract infection with influenza virus in human. Viral load measurement by semi-quantitative detection of viral RNA allows rapid and sensitive screening of potential compounds with in vitro anti-influenza A virus activity. PMID- 12719011 TI - Transcriptional suppression of the HIV promoter by natural compounds. AB - Tannins and lignins are natural compounds contained in plants such as tea leaves. Previously, we demonstrated that tannic acid represses 12-o-tetra-decanoyl phorbol-13-acetate (TPA)-induced human immunodeficiency virus (HIV) promoter activity. Furthermore, we demonstrated that a 30-bp element located just downstream of the NF-kappaB element in the HIV promoter responds negatively to tannic acid. However, the kinds of molecules responsible for this suppressive effect have remained unknown, because tannic acid is a mixture of various galloylglucoses. Here, we examined structure-defined natural compounds for HIV promoter-suppressive effects. We found that ellagitannins suppress TPA-induced HIV promoter activity to the same extent as tannic acid. 3-phenylcoumarins, isoflavone and chalcones have more suppressive effects than ellagitannins. On the other hand, other flavonoids and acetogenins have no suppressive effect. 3 phenylcoumarins and chalcones showed no suppressive effect on the cytomegalovirus (CMV) promoter, suggesting that they act specifically on the HIV promoter. These results suggest that 3-phenylcoumarin or chalcone compounds could be used to develop novel anti-HIV drugs with an action targeted at HIV promoter activity. PMID- 12719012 TI - Prevalence of Giardia spp. and Cryptosporidium spp. on dairy farms in southeastern New York state. AB - A prevalence study was designed to determine the presence of Cryptosporidium spp. and Giardia spp. in the soil of 37 dairy farms in southeastern New York state. A sampling design was developed and used to collect soil samples from these farms. Areas on the farms which were considered to be potential sources of contamination to the environment were evaluated quantitatively using a multidimensional scale. This scale included factors which could have the potential to contribute to the risk of contamination of the environment with Giardia or Cryptosporidium. In addition, the runoff pathway from these areas was identified and sampling points along that pathway were determined. Using a sampling grid, sampling sites were determined and soil samples collected and analyzed individually for the following: presence of Giardia and Cryptosporidium, pH, gravimetric moisture content, and volumetric moisture content. Out of 782 soil samples, 17% were positive for Cryptosporidium and 4% were positive for Giardia. The pH of the soil ranged from 3.7 to 9.8 with a mean of 7.0. There was a significant association between the pH and the likelihood of detecting Cryptosporidium spp. As the pH increased, the likelihood of detecting an oocyst decreased. Gravimetric moisture content had a mean of 40% and a range from 7 to 86%. There was a significant association between the gravimetric moisture content and the likelihood of detecting Giardia in soil samples. PMID- 12719013 TI - Oestrus ovis larval myiasis among goats in northern Jordan. AB - From December 1998 to December 1999, heads of 520 local goats slaughtered at the Irbid, Ramtha and Howarra Abattoirs (northern Jordan) were examined for the three larval instars (L(1)-L(3)) of Oestrus ovis. Of 520 heads, 126 (24%) were infested with O. ovis larvae. All three larval instars were observed in both sexes; all age groups were infested in each month of the year. The mean age of the goats sampled was 1.5 years. The numbers of parasites infesting hosts showed a significant (P<0.05) correlation with sheep age (r(sp)=0.31-0.42) for all three larval instars. The numbers of larvae in each host followed an overdispersed distribution, which fit a negative-binomial model (but not a Poisson distribution). There were more parasites recorded in the presence of purulent discharge or laryngitis, fewer in the presence of catarrhal discharge and no association with pharyngitis sinusitis, or rhinitis. PMID- 12719014 TI - Sheep blowfly strike reduction using a synthetic lure system. AB - The effectiveness of a synthetic lure system to reduce the incidence of blowfly strike in sheep flocks was assessed, using randomised field trials. Four field trials used eight total groups of sheep randomised to treatment (flytrap) or control on two properties in southern Queensland between 1999 and 2001. Treatment consisted of the operation of flytraps in paddocks as per manufacturer's recommendations. All sheep were inspected physically each month for flystrikes. Flytraps were associated with a reduction in flystrike incidence of 38-55%, compared to control sheep. Results confirm that traps are a useful component of a flystrike-control program. The use of fly traps by a substantial proportion of woolgrowers could assist the Australian wool industry to meet targets of reduced pesticides on shorn wool. PMID- 12719015 TI - A decision-support system for Salmonella in broiler-chicken flocks. AB - We built a decision-support system to assess the risk of contamination of chicken broiler flocks by Salmonella at the end of the rearing period. This system was developed from the survey data from 85 chicken-broiler flocks located in western France. First, we estimated the probability of contamination of the house by Salmonella before placement of day-old chicks via a cleansing inspection using a visual-inspection grid, a decontamination evaluation using count-plates, and risk factors for Salmonella persistence in the barn after cleansing and disinfection. Second, we estimated (using a logistic model) the probability of prevalent contamination of the flock by Salmonella at the end of the rearing period. Validation was carried out on 60 flocks selected from seven production companies in western France. The risk estimated by the model was compared to the Salmonella status of the flock (gold standard) assessed by samples taken from the environment of the broilers and analysed with classical bacteriological methods. The sensitivity was 97.8% and the specificity 64.3%. PMID- 12719016 TI - Eradication of enzootic bovine leukosis from Finland. AB - Enzootic bovine leukosis (EBL) was recognized among Finnish cattle in 1966. Administrative decisions specifying and refining official control measures were given in 1966, 1976, 1980, and 1993. The measures' key principle always has been 'test and slaughter'. The EBL/bovine leukosis virus (BLV) infection situation was monitored at meat inspection, and hematologically between 1970 and 1977 and serologically between 1978 and 1989. Annual surveys including all dairy herds and samples from beef animals were conducted in 1990-2001. Bulk-tank milk samples represented the dairy herds in the surveys; the beef animals were sampled individually at slaughter. The maximum positive herd-level percentage in the surveys was 0.03%. EBL/BLV infection was evenly dispersed in the southern part of the country and nonexistent in the northern part. We conclude that herd-level prevalence of EBL/BLV infection never exceeded 5%. It nevertheless took 30 years to eradicate the disease and the infection. EBL was eradicated from mainland Finland in 1996 and from the island district of Ahvenanmaa in 1999. Annual monitoring of the EBL situation continues. PMID- 12719017 TI - A linear programming approach to estimate the economic impact of bovine viral diarrhoea (BVD) at the whole-farm level in Scotland. AB - We combined epidemiological and economic concepts and modelling techniques, to integrate animal health into whole-farm business management. This allowed us to assess the relative contribution that disease prevention could make to whole-farm income and to the variability in farm income (risk). It also allowed us to assess disease losses in the context of a farm business rather than as a disease outbreak in isolation. A linear program ("MOTAD") establishes the combination of decision maker's activities that minimise risk for a given level of income within farm-business constraints. The MOTAD model was applied to farm-management decision making in Scottish cow-calf herds and was linked to an epidemiological model of bovine viral diarrhoea (BVD). When BVD was considered in isolation (i.e. without taking into account risk), the minimum expected total cost of BVD (sum of output losses plus expenditure on prevention) was similar whether the herd was susceptible to BVD or of unknown BVD-status at the outset. However, the expected total cost of BVD fell in response to increasing expenditure on prevention in 'susceptible' herds. This relationship was not apparent in herds of unknown BVD status. As a consequence of this difference, 'susceptible' herds were better able to use investment in BVD biosecurity as a means to increase farm income at minimum risk than herds of unknown BVD-status. 'Susceptible' herds therefore were able to achieve high income targets with less-intensive production than herds of unknown BVD-status. This suggested that maintaining a cow-calf herd free of BVD contributes to farm income and risk management indirectly through its effect on the management of the whole farm. It follows that measurement of the economic impact of BVD requires a whole-farm perspective that includes a consideration of risk. Because farmers generally are considered to be risk adverse, this means that the least-cost disease-control option might not always be the preferred option. PMID- 12719018 TI - Prevalence of and carcass condemnation from maedi-visna, paratuberculosis and caseous lymphadenitis in culled sheep from Quebec, Canada. AB - We determined the prevalence of lung and mammary gland lesions associated with maedi-visna (MV) infection, the prevalence of paratuberculosis (PTB), and the prevalence and lesions distribution of caseous lymphadenitis (CL) in culled sheep. Total of 451 ewes and 34 rams were selected randomly from two slaughterhouses in Quebec, Canada. MV serostatus was determined by recombinant ELISA test. PTB diagnosis was based on characteristic histological lesions in the terminal ileum, ileocecal lymph node and/or ileocecal valve and CL by gross detection of abscesses and isolation of Corynebacterium pseudotuberculosis. Seroprevalence of MV was 44% (95% CI: 40, 48). Seropositivity increased with age and was higher in ewes than in rams. The percentages of lung and mammary gland lesions in seropositive sheep were 14 and 40%, respectively, but mammary gland lesions lack specificity. The prevalence of PTB was 3% (95% CI: 2, 5). PTB increased with age and was lower among sheep with abscesses. The prevalence of CL was >/=21% (95% CI: 17, 24). The most-prevalent site of caseous lymphadenitis lesions was the thoracic cavity. The risk of carcass condemnation was significantly associated with region, body score and abscesses. Only the presence of abscesses was associated with an increase in trimming of carcasses. PMID- 12719019 TI - The effect of non-nutritional factors on milk urea nitrogen levels in dairy cows in Prince Edward Island, Canada. AB - We determined the effects of non-nutritional factors such as breed, parity, days in milk (DIM), milk production, milk quality and milk components on milk urea nitrogen (MUN) concentration. A total of 177 dairy farms in Prince Edward Island containing 10,688 lactating dairy cows participated in the project. Individual cow milk samples (n=68,158) were collected monthly from July 1999 to June 2000 from each farm. MUN was measured using a Fossomatic 4000 Milkoscan Analyzer at the PEI Milk Quality Laboratory. Descriptive statistics for MUN, parity, DIM, and test-day milk yield, fat and protein were calculated. Mixed linear-regression models were used; "cow" and "herd" were included as random effects to control for the effect of clustering of MUN test dates within cow, and clustering of cows within herd, respectively. The MUN was lower during the first month of lactation, peaked at 4 months of lactation, and decreased later in lactation. A positive relationship existed between MUN concentration and milk yield, while negative relationships with milk protein% and linear score were observed. A quadratic relationship existed between milk fat% and MUN concentration, with higher MUN occurring at mid-range fat percentages. The variation at the herd and cow levels in the model were 19.7 and 19.0%, respectively; while the variation at the test date level was 61.3%. The non-nutritional factors studied explained 13.3% of the variation in MUN. PMID- 12719020 TI - A clinical trial of the effect of a mouth-rinse prepared with Lippia sidoides Cham essential oil in dogs with mild gingival disease. AB - We evaluated the effect of a mouth-rinse prepared using Lippia sidoides essential oil (EO) in dogs with marginal gingivitis. German Shepherd dogs were distributed in two groups: control (control mouth-rinse) and EO (EO mouth-rinse). Both mouth rinses were applied on the dogs' teeth every 2 days for 2 weeks. At day 0 and day 15, the scores for plaque-bacteria (P), calculus (C), gingivitis (G) and the inflammatory infiltrate (INF) were evaluated blindly. The results were analyzed by the Wilcoxon signed-rank and Mann-Whitney tests (P 1 microg/L (n = 8); Group 2, FCp < 1 microg/L and >0.45 microg/L (n = 9); and Group 3, FCp < 0.45 microg/L (n = 11). PCp, BIS, recovery time, and other data were compared between the three groups. MAIN RESULTS: Demographic values, duration of surgery, and consumption of propofol and fentanyl were not different between the three groups. Group 3 patients regained consciousness with significantly higher propofol concentration (mean PCp = 3.2 mg/L) compared with those in Groups 1 and 2 (p < 0.05). However, the BIS values at both recovery endpoints were not different among the three groups. CONCLUSIONS: The plasma levels of fentanyl affect the concentrations of propofol required for patients to regain consciousness. The BIS values for wakefulness are unaltered at the different combinations of propofol and fentanyl concentrations. Thus, the BIS appears to be a useful and consistent indicator for level of consciousness during emergence from propofol/fentanyl intravenous anesthesia. PMID- 12719049 TI - How much are patients willing to pay to avoid intraoperative awareness? AB - STUDY OBJECTIVE: To determine how much patients are willing to pay to avoid intraoperative awareness? DESIGN: Observational study SETTING: University affiliated metropolitan hospital. PATIENTS: 60 patients who completed a questionnaire (39 F, 21 M). The mean age was 43 years and the median household income range of 45,000-60,000 US dollars. INTERVENTIONS: Patients completed an interactive computer-generated questionnaire on the value of preventing intraoperative awareness and their willingness to pay for a "depth of anesthesia" monitor. Their willingness to pay for the prevention of postoperative pain, nausea and vomiting, postoperative grogginess, and sleepiness was also determined as a means of comparison. MEASUREMENTS AND MAIN RESULTS: Patients were willing to pay (WTP) 34 US dollars, 10 US dollars to 42 US dollars (median, interquartile range) for a monitor that would assist an anesthesia care provider assess the depth of anesthesia in an effort to avoid awareness. This increased to 43 US dollars, 20 US dollars to 77 US dollars (p < 0.0,001) (median, interquartile range), if the insurance company was making the payment and the WTP value only decreased minimally to 33 US dollars if the incidence of awareness was reduced 10 fold. CONCLUSION: The incidence of intraoperative awareness and WTP value for monitoring awareness have a nonlinear relationship (a risk averse utility function), which suggests that patients assign an intrinsic base value for a rare or very rare possibility of an event. Other healthcare economic analyses (such as cost effectiveness) do not take this factor into account and assume a linear value relationship (i.e., if something occurs ten times less frequently, it has ten times less value). IMPLICATION: The median value for patients' WTP for a monitor that might prevent awareness under anesthesia was 34 US dollars given an incidence of 5/1,000 cases. The incidence of awareness and WTP value have a nonlinear relationship suggesting that patients assign an intrinsic base value for the possibility of awareness. PMID- 12719050 TI - Wound infiltration with ropivacaine and fentanyl: effects on postoperative pain and PONV after breast surgery. AB - STUDY OBJECTIVE: To determine whether postoperative wound infiltration with local anesthetics combined with fentanyl improves analgesia following breast surgery; and to investigate awakening and postoperative nausea/vomiting. DESIGN: Prospectively randomized clinical study. SETTING: University hospital. PATIENTS: 45 ASA physical status I and II patients scheduled for breast surgery. INTERVENTIONS: Patients were prospectively randomized and assigned to one of three treatments during general anesthesia: postsurgical wound infiltration with ropivacaine 0.375%; wound infiltration with ropivacaine 0.375% combined with fentanyl 0.5 microg/kg; and intravenous (i.v.) fentanyl 0.5 microg/kg before skin incision and no wound infiltration. Time to first verbal response, pain at rest, postoperative nausea and vomiting, and ketobemidone and dixyrazine utilization were compared. MEASUREMENTS AND MAIN RESULTS: Time to first verbal response was significantly shorter in the i.v. fentanyl group compared to both infiltration groups (8.1 +/- 4.5 min vs. 15.3 +/- 4.3, and 12.0 +/- 5.0 min; p < 0.05, respectively). Postoperative pain at rest, and nausea and vomiting occurred with similar frequencies in the groups. Ketobemidone utilization in both infiltration groups, (2.4 +/- 1.8 mg and 3.1 +/- 1.8 mg, respectively) was not different compared to the i.v. fentanyl group (2.9 +/- 2.0 mg; NS). There were no differences in postoperative antiemetic requirements during the first, second and third two-hour periods postoperatively. The dixyrazine consumption was similar in the three groups, (0.9 +/- 1.5 mg, 0.8 +/- 1.3 mg, and 1.4 +/- 1.8 mg, respectively; NS). CONCLUSION: Postsurgical ropivacaine wound infiltration, with or without adding fentanyl, demonstrates no differences in postoperative pain relief and nausea/vomiting compared to a balanced general anesthetic including i.v. fentanyl. PMID- 12719051 TI - Preoperative combined with intraoperative skin-surface warming avoids hypothermia caused by general anesthesia and surgery. AB - STUDY OBJECTIVES: To evaluate the effects of intraoperative skin-surface warming with and without 1 hour of preoperative warming, in preventing intraoperative hypothermia, and postoperative hypothermia, and shivering, and in offering good conditions to early tracheal extubation. DESIGN: Prospective, randomized, blind study. SETTING: Teaching hospital. PATIENTS: 30 ASA physical status I and II female patients scheduled for elective abdominal surgery. INTERVENTIONS: Patients received standard general anesthesia. In 10 patients, no special precautions were taken to avoid hypothermia. Ten patients were submitted to preoperative and intraoperative active warming. Ten patients were only warmed intraoperatively. MEASUREMENTS AND MAIN RESULTS: Temperatures were recorded at 15-minute intervals. The patients who were warmed preoperatively and intraoperatively had core temperatures significantly more elevated than the other patients during the first two hours of anesthesia. All patients warmed intraoperatively were normothermic only at the end of the surgery. The majority of the patients warmed preoperatively and intraoperatively or intraoperatively only were extubated early, and none had shivering. In contrast, five unwarmed patients shivered. CONCLUSIONS: One hour of preoperative warning combined with intraoperative skin surface warming, not simply intraoperative warming alone, avoided hypothermia caused by general anesthesia during the first two hours of surgery. Both methods prevented postoperative hypothermia and shivering and offered good conditions for early tracheal extubation. PMID- 12719052 TI - Intraoperative epidural anesthesia and postoperative analgesia with levobupivacaine for major orthopedic surgery: a double-blind, randomized comparison of racemic bupivacaine and ropivacaine. AB - STUDY OBJECTIVE: To compare the onset time and duration of epidural anesthesia, and the quality of postoperative analgesia produced by levobupivacaine, racemic bupivacaine, and ropivacaine. DESIGN: Prospective, randomized, double-blinded study. SETTING: Inpatient anesthesia at a University Hospital. PATIENTS: 45 ASA physical status I, II, and III patients, undergoing elective total hip replacement. INTERVENTIONS: After standard intravenous midazolam premedication and infusion of 500 mL of Ringer's acetate solution, patients were randomly allocated to receive epidural block with 0.5% levobupivacaine (n = 15), 0.5% bupivacaine (n = 15), or 0.5% ropivacaine (n = 15). Postoperatively, after pinprick sensation recovered at T(t), a patient-controlled epidural infusion was provided with 0.125% levobupivacaine, 0.125% bupivacaine, or 0.2% ropivacaine, respectively (baseline infusion rate 5 mL/hr; incremental bolus 2 mL, lockout time: 20 min). Intravenous ketoprofen was also available for rescue analgesia if required. MEASUREMENTS AND MAIN RESULTS: The onset time of sensory block was 31 +/- 16 minutes with levobupivacaine, 25 +/- 19 minutes with bupivacaine, and 30 +/- 24 minutes with ropivacaine (p = 0.98), after a median (range) volume of 15 (10-18) mL in Group Levobupivacaine, 14 (10-18) mL in Group Bupivacaine, and 15 (10-18) mL in Group Ropivacaine (p = 0.85). Six patients in the ropivacaine group (40%) showed an intraoperative Bromage score <2 as compared with only three patients of Group Levobupivacaine (20%) and no patient of Group Bupivacaine (p = 0.02). Recovery of pinprick sensation at T(t) occurred after 214 +/- 61 minutes with levobupivacaine, 213 +/- 53 minutes with bupivacaine, and 233 +/- 34 minutes with ropivacaine (p = 0.26). A similar degree of pain relief was observed in the three groups without differences in local anesthetic consumption and need for rescue analgesia. Motor blockade progressively resolved without differences among the three groups. CONCLUSIONS: Levobupivacaine 0.5% produces an epidural block of similar onset, quality, and duration as the one produced by the same volume of 0.5% bupivacaine, with a motor block deeper than that produced by 0.5% ropivacaine. When prolonging the block for the first 12 hours after surgery with a patient-controlled epidural infusion, 0.125% levobupivacaine provides adequate pain relief after major orthopedic surgery, with similar recovery of motor function as compared with 0.125% bupivacaine and 0.2% ropivacaine. PMID- 12719053 TI - Hemodynamic and catecholamine response to tracheal intubation: direct laryngoscopy compared with fiberoptic intubation. AB - STUDY OBJECTIVE: To compare the stress response following tracheal intubation using direct laryngoscopy to that using fiberoptic bronchoscopy technique. DESIGN: Randomized, prospective study. SETTING: Operating rooms in a teaching hospital. PATIENTS: 51 ASA physical status I and II patients who were scheduled for an elective surgery with general anesthesia. INTERVENTIONS: Patients were randomly assigned to receive either direct laryngoscopy or fiberoptic orotracheal intubation, as part of general anesthesia. A uniform protocol of anesthetic medications was used. MEASUREMENTS: Blood pressure and heart rate were measured before induction, before endotracheal intubation, and 1, 2, 3, and 5 minutes afterwards. Catecholamine (epinephrine and norepinephrine) blood samples were drawn before the induction, and 1 and 5 minutes after intubation. MAIN RESULTS: Duration of intubation was shorter in the direct laryngoscopy group (16.9 (16.9 +/- 7.0 sec, range 8 to 40) compared with the fiberoptic intubation group (55.0 +/- 22.5 sec, range 29 to 120), p < 0.0,001. In both groups, blood pressure and heart rate were significantly increased at 1, 2, and 3 minutes after intubation, but there was no significant difference between the two study groups. Catecholamine levels did not increase after intubation and did not correlate with the hemodynamic changes. CONCLUSIONS: The use of either direct laryngoscopy or fiberoptic bronchoscopy produces a comparable stress response to tracheal intubation. Catecholamine levels do not correlate with the hemodynamic changes. PMID- 12719054 TI - Another cause of difficulty in ventilating a patient. AB - Various mechanical problems with the delivery of anesthesia gases have been reported, and preoperative checks of the anesthesia delivery system are designed to prevent these. We report a case of a mechanical obstruction of the expiratory limb of the anesthesia circuit which went undetected before the induction of anesthesia. A check of the circuit was performed before the addition of circuit extensions, thus missing the obstruction. Systematic investigation of the anesthetic system after difficulty in ventilating the patient revealed the obstruction, without any harmful consequence to the patient. This case highlights the importance of a thorough check of the system before any anesthetic, particularly after any change in the circuit. PMID- 12719055 TI - Difficult removal of a wire-reinforced epidural catheter. AB - We report a case of epidural catheter shearing, an uncommon complication of removal of a wire-reinforced epidural catheter. The catheter was removed without incident, and with the tip intact, after we allowed the patient to relax for 3 hours, placing the patient in the lateral decubitus position, and placing continuous tension on the catheter itself so as to let it "work its way out". PMID- 12719056 TI - Transpleural subclavian central venous catheter placement in a child with scoliosis discovered during a thoracotomy. AB - Placement of central venous catheters in dysmorphic children can be difficult because of distortion of normal anatomical landmarks. We present such a case of a 16 year-old child who had a central venous catheter inserted in the left subclavian vein. Although a conventional roentgenogram was consistent with correct placement, the catheter was found to traverse the pleural space before entering the subclavian vein. PMID- 12719057 TI - Acceleromyography as a guide to anesthetic management: a case report. AB - We present a case of prolonged recovery from mivacurium. Neuromuscular monitoring using acceleromyography was extremely helpful following attempted reversal of residual block in determining when tracheal extubation could be safely performed. If a method of objective estimation of the TOF ratio had not been available, tracheal extubation would have taken place at a time when the train-of-four fade ratio was below 0.40. PMID- 12719058 TI - Instruction of airway management skills during anesthesiology residency training. AB - BACKGROUND: Difficult airway management is a critical aspect of anesthesiology training and practice. A survey was conducted of American anesthesia residency programs to determine the prevalence of a specific airway rotation and its curriculum. METHODS: A questionnaire was sent by both e-mail and fax to all 132 directors of American anesthesiology residency programs. RESULTS: Of the 132 programs surveyed, 79 (60%) responded. Of the responders, 26 programs (33%) have a difficult airway rotation. The rotation was offered throughout the years of clinical training in 13 (49%) of the programs and was of 1-week duration in 16 (61%) of these programs who had a rotation. Formal instruction was administered before the rotation in 18 (69%) of the programs. Instruction was usually performed on surgical patients in 22 (85%) of the program, ASA status I and II patients in 20 (77%) of the programs, and taught by selected faculty in 20 (78%) of the programs. There was a 2- to 5-minute time limitation or a number of maximum attempts when using any of these devices in 16 (62%) programs. There was a case number requirement regarding the use of the devices in 5 (19%) of the programs. Residents were evaluated by both skills testing and written evaluation in 63% of these programs. DISCUSSION: Of the programs with an airway rotation, the flexible fiberoptic bronchoscope and the laryngeal mask airway represent the most frequently used devices, excluding the standard laryngoscopes. There is minimum emphasis on more invasive techniques. Traditional methods of instruction continue to be utilized more frequently than nontraditional methods. PMID- 12719059 TI - Norepinephrine in septic patients--friend or foe? AB - Norepinephrine (NE) is mostly used to treat severe hypotension. However, NE has potentially adverse vasoconstrictive effects on regional vascular beds of kidney, liver, and gut, with a potential for ensuing organ dysfunction. NE therefore is considered as a last reserve in otherwise refractory hypotension. During sepsis, a loss of catecholamine responsiveness occurs that is often interpreted as down regulation of catecholamine receptors. Therefore, the doses of NE needed to maintain or restore blood pressure may be extremely high. Surprisingly, no adverse vasoconstriction with subsequent hypoperfusion occurs during sepsis, despite the high doses of NE administered. Instead, NE rather causes an increase in blood flow and oxygen delivery. PMID- 12719060 TI - Postoperative atelectasis after one-lung ventilation with the Univent tube in a child. AB - This case describes a postoperative complication after single-lung ventilation in a 10-year-old patient scheduled for scoliosis repair. This patient underwent an anterior thoracoscopic excision of several vertebral discs followed by a posterior double-rod fixation of her spine. The postoperative course was complicated by complete atelectasis of the patient's right lung for 5 days. Methods of single-lung ventilation, advantages and disadvantages of the Univent tube versus the double-lumen tube, and the possible mechanisms leading to postoperative atelectasis in this child are discussed. PMID- 12719061 TI - The pinch test: a simple technique for determining the pressure of an endotracheal tube cuff. PMID- 12719062 TI - Correlation of visual and refractive outcomes between eyes after same-session bilateral laser in situ keratomileusis surgery. AB - PURPOSE: To determine whether between-eye refractive and visual outcomes after same-session laser in situ keratomileusis (LASIK) surgery are correlated, and to determine whether suboptimal visual and refractive outcomes in one eye are predictive of poor results in the fellow eye. DESIGN: Observational case series. METHODS: Retrospective chart review. A total of 484 eyes of 242 patients met inclusion criteria of having undergone same-day LASIK surgery and having 3-month refractive and visual acuity outcomes. Statistical comparisons of outcomes were performed, including between-eye Pearson correlation analysis and logistic regression models for predicting second-eye outcome. Main outcome measures were 1 month, 3-month, and 6-month uncorrected visual acuity and manifest refraction. RESULTS: Refractive outcomes at 1, 3, and 6 months between first (right) and second eyes were not found to be significantly different. No difference in uncorrected or best spectacle-corrected visual acuity was observed between first and second eyes Refractive outcomes of spherical equivalent, sphere, and cylinder and visual outcomes of uncorrected and best spectacle-corrected visual acuity were highly correlated between the two eyes. Analysis of cases with suboptimal outcomes (> or =1 diopter from intended correction or uncorrected acuity equal to or worse than 20/40) suggests that a poor refractive or visual outcome in the first eye increases the risk of a poor outcome in the second eye by approximately 20-fold. CONCLUSIONS: Retrospective analysis of refractive and visual outcomes from patients undergoing same-session bilateral LASIK demonstrates a high correlation of refractive and visual outcome between the eyes. PMID- 12719063 TI - Increased endothelial cell density in the paracentral and peripheral regions of the human cornea. AB - PURPOSE: To systematically investigate the central, paracentral, and peripheral endothelial cell density (ECD) in normal human corneas. DESIGN: Observational case series and experimental study. METHODS: Noncontact specular microscopy was undertaken to determine the ECD of the central, paracentral (2.7 +/- 0.2 mm from center) and peripheral (4.7 +/- 0.2 mm from center) regions of the cornea of 48 normal eyes. The ECDs of central and peripheral regions were also determined with contact specular microscopy in 21 normal eyes and a group of 30 Optisol-GS eye bank corneas were evaluated with alizarin red stain. Histologic ECD of 13 Optisol GS stored corneas were also determined. RESULTS: Paracentral and peripheral ECD measured with the noncontact specular microscope were 5.8% (P <.01) and 9.6% (P <.001) increased compared with central ECD. Superior peripheral ECD was increased compared with the other three peripheral quadrants (P <.05) and was 15.9% higher than central ECD. Contact specular microscopy showed an increase of 8.9% in the peripheral ECD from the center. Alizarin red stained corneas confirmed the specular microscopy numbers with a 9.2% increase in the paracentral region, and a 17.2% increase in the peripheral region. Histological cross sections of human corneas also showed a 22.9% increase in peripheral ECD compared with the central region. CONCLUSIONS: The human cornea has an increased ECD in the paracentral and peripheral regions of cornea compared with the central region. The superior peripheral region of the corneal endothelium has the largest increase in ECD. These data on normal endothelial cell distribution in the human cornea are especially significant as they relate to new surgical techniques and endothelial wound repair. PMID- 12719064 TI - Proposed pathogenesis for the delayed postoperative opacification of the hydroview hydrogel intraocular lens. AB - PURPOSE: To report the clinical, histopathologic, ultrastructural, and elemental features of 17 opacified Hydroview (Bausch and Lomb Surgical, Rochester, New York, USA) hydrogel intraocular lenses (IOL) necessitating explantation and discuss from a clinicopathologic perspective why these lenses became opacified. Interventional case series with clinicopathologic correlation. METHODS: Seventeen hydrogel lenses were explanted from 17 different patients owing to decreased visual acuity or quality of vision an average of 29 months after uneventful phacoemulsification and IOL implantation and associated with a granular-appearing opacification superficially within the optic. Lenses were examined by light microscopy, transmission electron microscopy (TEM), and energy dispersion x-ray (EDX) spectroscopy. A control IOL was included in our study. RESULTS: All explanted lenses showed positive staining for calcium by light microscopy. Transmission electron microscopy disclosed electron-dense crystalline deposits in the superficial substance of the IOL optic. Energy dispersion x-ray spectra analyses showed the presence of calcium and phosphorus mainly in the electron dense periphery of the deposits in all of the specimens and the presence of silicon mainly in the electron-lucent center of the deposits in the majority of the specimens. No positive staining or deposits were observed on the IOL control or in the haptics. CONCLUSIONS: Our study is the first to demonstrate that the calcium deposits are associated with silicon, which was presumably derived from the silicone gasket in the Surefold (Bausch and Lomb Surgical, Rochester, New York, USA) packaging system, manufactured specifically for this IOL. Silicon may act as a nidus for calcium deposition within the lens, which is consistent with our findings. There may be other factors involved, and this important clinical problem requires further study. PMID- 12719065 TI - A self-administered health questionnaire for the preoperative risk stratification of patients undergoing cataract surgery. AB - PURPOSE: To determine if a self-administered health status questionnaire completed by candidates for cataract surgery is beneficial for identifying medical comorbidities and patients at risk for adverse intraoperative and postoperative medical events. DESIGN: Prospective cohort study. METHODS: Data were obtained from a large, randomized clinical trial of 19,250 cataract surgeries performed between June 1, 1995, and June 30, 1997. Preoperative data were obtained from a standardized, self-administered patient health questionnaire and a history and physical form completed by the patient's physician. A record of adverse medical events on the day of surgery and through the first 7 postoperative days was recorded. RESULTS: Responses to 21 questions on the questionnaire were highly specific for 12 comorbid conditions identified by the physician history and physical, ranging from 100% specificity for arrhythmia to 91.3% specificity for coronary artery disease. Comorbid conditions identified by the questionnaire were associated with similar relative risks of adverse events as those identified by the physician history and physical. Patients who had no comorbidities identified by the questionnaire had adverse medical event rates indistinguishable from patients with no comorbid conditions identified by the health provider history and physical. CONCLUSIONS: The self-administered questionnaire showed a high degree of specificity for 12 common comorbid conditions in cataract patients. A questionnaire such as this may be useful in preoperative risk stratification. PMID- 12719066 TI - A new method for tear film stability analysis using videokeratography. AB - PURPOSE: To report a new tear film stability analysis system using videokeratography. DESIGN: Observational case series. METHODS: New videokeratography software for TMS-2N (topographic modeling system; TOMEY Corporation, Nagoya, Japan) was developed that can automatically capture consecutive corneal surface images every second for 10 seconds. Forty-eight adult volunteers (80 eyes) were recruited for this study, and all subjects were examined with the new system. Corneal topographs were analyzed for tear breakup time (TMS-BUT) and the ratio of breakup area to entire color-code area (TMS-BUA) was calculated. Routine methods for tear film breakup time evaluation using slit lamp microscope and fluorescence staining (SLE-BUT) were performed for comparison purposes. Regressive correlations of TMS-BUT or TMS-BUA with SLE-BUT were analyzed. Based on SLE results, subjects were separated into two groups with normal and short BUT, respectively. TMS-BUT and TMS-BUA were compared with SLE BUT data with regard to the sensitivity and specificity of evaluation of dry eye symptoms. RESULTS: Topographic modeling system-tear breakup time (TMS-BUT) had a positive correlation with SLE-BUT (R = 0.7219, P <.0001), whereas TMS-BUA showed a negative correlation (R = 0.6317, P <.0001). Among 34 eyes with normal SLE-BUT, 11 eyes (32.35%) displayed abnormal TMS-BUT, 9 (81.82%) of which were associated with dry eye symptoms. The sensitivities of TMS-BUT and TMS-BUA were 97.5% and 95%, respectively, significantly higher than that of SLE-BUT (75%), with P =.008 and 0.01, respectively. Topographic modeling system-BUT and TMS-BUA displayed a similar rate of specificity in comparison with SLE-BUT. CONCLUSIONS: This new videokeratography system is a noninvasive and objective method with increased sensitivity for tear film stability analysis. PMID- 12719067 TI - Lecithin-bound superoxide dismutase in the treatment of noninfectious corneal ulcers. AB - PURPOSE: To evaluate the effects of a lipophilic analog of superoxide dismutase in the treatment of persistent epithelial defects of the cornea. DESIGN: Interventional case series. METHODS: Nineteen eyes of 19 patients with noninfectious corneal ulcers were enrolled. Patient profiles consisted of seven cases with Mooren-type peripheral ulcers, three cases with chemical or thermal burns, two cases with vernal keratoconjunctivitis, four cases with cicatricial keratoconjunctivitis, and three cases with sterile ulcers after corneal surgery. Patients consisted of 11 men and 8 women, with a mean age of 51.5 +/- 17.9 years (range, 14-77). Only patients who did not respond to conventional therapy were recruited for the study. Lecithinated superoxide dismutase was applied for 2 weeks, and sketches and/or fluorescein slit micrographs were recorded before and at the end of the study. The main outcome measure was decrease in ulcer size. RESULTS: As a group, the average ulcer size relative to the entire cornea decreased from 18.2 +/- 15.1% to 10.0 +/- 15.0% after 2 weeks (P <.05). A dramatic effect (>90% decrease in ulcer size) was observed in eight cases (100% in six cases), whereas no apparent change was found (<30% decrease in ulcer size) in five cases. The presence of polymorphonucleocytes within the ulcerated tissue was confirmed by brush cytology. CONCLUSION: Lecinthinated superoxide dismutase was effective in decreasing persistent epithelial defect size in a subset of patients with sterile corneal ulcers refractive to conventional therapy. A large scale clinical investigation is merited. PMID- 12719068 TI - Riboflavin/ultraviolet-a-induced collagen crosslinking for the treatment of keratoconus. AB - PURPOSE: In animal eyes, a significant increase in corneal biomechanical stiffness has been found after collagen crosslinking by combined riboflavin/ultraviolet-A (UVA) treatment. The aim of the present study was to evaluate the clinical usefulness of riboflavin/UVA-induced collagen crosslinking for bringing the progression of keratoconus to a halt. DESIGN: Prospective, nonrandomized clinical pilot study. METHODS: Twenty-three eyes of 22 patients with moderate or advanced progressive keratoconus (maximum K value, 48-72 diopters) were included. After central corneal abrasion, photosensitizing riboflavin drops were applied and the eyes exposed to UVA (370 nm, 3 mW/cm(2)) in a 1-cm distance for 30 minutes. Postoperative examinations were performed in 6 month intervals, including visual acuity testing, corneal topography, slit-lamp examination, measurement of endothelial cell density, and photographic documentation. The follow-up time was between 3 months and 4 years. RESULTS: In all treated eyes, the progression of keratoconus was at least stopped. In 16 eyes (70%) regression with a reduction of the maximal keratometry readings by 2.01 diopters and of the refractive error by 1.14 diopters was found. Corneal and lens transparency, endothelial cell density, and intraocular pressure remained unchanged. Visual acuity improved slightly in 15 eyes (65%). CONCLUSIONS: Collagen crosslinking may be a new way for stopping the progression of keratectasia in patients with keratoconus. The need for penetrating keratoplasty might then be significantly reduced in keratoconus. Given the simplicity and minimal costs of the treatment, it might also be well-suited for developing countries. Long-term results are necessary to evaluate the duration of the stiffening effect and to exclude long term side-effects. PMID- 12719069 TI - Myopic astigmatism and presbyopia trial. AB - PURPOSE: No prospective double-masked study has evaluated whether low astigmatism benefits or harms patients with presbyopia, whose intermediate and near vision might theoretically benefit from enhanced depth of focus provided by astigmatism. The purpose of the first Myopic Astigmatism and Presbyopia (MAP I) study was to determine whether low myopic astigmatism enhances or harms the visual acuity, stereopsis, or quality of life in patients with presbyopia. DESIGN: Prospective, randomized, double-masked, crossover design clinical trial. METHODS: Fifteen patients with presbyopia aged 45 to 68 years were recruited from an academic center population. These patients were given a baseline eye examination, including manifest refraction, Early Treatment of Diabetic Retinopathy Study (ETDRS) logarithm of minimal angle of resolution (logMAR) visual acuity at distance, intermediate, and near, accommodative amplitudes, and stereo vision. Each patient was then cycled in random order through three masked pairs of soft contact lenses. The power of each contact lens pair was calculated by the subtraction method to maintain a spherical equivalent of -0.5 diopters, while providing either no astigmatism (spherical arm, SPH), 1 diopter of with-the-rule (WTR) astigmatism, or 1 diopter of against-the-rule (ATR) astigmatism. Actual refractive errors produced were measured by masked examiner. Outcomes measured at the end of 1 week of usage of each contact lens arm were binocular (ETDRS) logMAR visual acuity at three distances (far [4 m], intermediate [1 m], and near [33cm]); near stereoacuity, using the quantitative Titmus Stereotest; and quality of life, measured using the Refractive Status and Vision Profile (RSVP), a standardized questionnaire. RESULTS: Visual acuity results across the three arms were similar. However, 1-m logMAR visual acuity was better for the spherical arm than either astigmatic arm (-0.06 SPH, +0.01 WTR, +0.02 ATR). Near (33 cm) and distance (4 m) acuities were similar across arms. Stereoacuity was better in ATR than WTR (50 vs 102 seconds, P =.01). Subjects preferred SPH slightly over the WTR astigmatic arm by the RSVP quality-of-life survey instrument (101 vs 104, P =.05). Other intergroup comparisons showed no difference in RSVP scores. CONCLUSIONS: This study has demonstrated that intermediate distance acuity and refractive quality of life are slightly better with spherical low myopic refractive error vs either astigmatic arm. Near and far distance acuity were unaffected by low myopic astigmatism compared with spherical low myopia. Near stereopsis was best in the ATR arms, but this did not produce better near visual acuity or RSVP quality of life. PMID- 12719070 TI - Long-term anatomic and visual acuity outcomes after initial anatomic success with macular hole surgery. AB - PURPOSE: To investigate the anatomic and visual outcomes in patients with initial anatomic success after macular hole surgery and with at least 5 years of follow up. DESIGN: Retrospective, noncomparative, consecutive case series. METHODS: Medical records of all patients who underwent surgery for idiopathic full thickness macular holes by two surgeons (W.E.S., H.W.F.) at the Bascom Palmer Eye Institute between January 1, 1991, and December 31, 1996, were reviewed. All patients who had initial anatomic success with macular hole surgery and who had 5 years or more of follow-up postoperatively were included in the study. Main outcome measures included the rate of macular hole reopening and visual acuity outcomes. RESULTS: Seventy-four eyes of 66 patients with a median age of 68.0 years (range, 45.0-86.8 years) were identified. The median duration of macular hole was 6.0 months (range, 1.1-93.8 months), and the median duration of follow up after macular hole surgery was 91.0 months (range, 60.0 to 114.8 months). The hole reopened in 9 eyes (12%) during the follow-up interval; 6 of these eyes underwent reoperation, and the hole closed in 4 of 6 (67%). Preoperative visual acuity ranged from 20/50 to 20/400 (mean, 20/129; median, 20/100). In the 62 eyes that underwent cataract extraction (CE) after macular hole surgery, CE was performed at a median of 13.9 months after macular hole surgery. Patients achieved their best postoperative visual acuity at a median of 28.5 months after macular hole surgery. Best postoperative visual acuity ranged from 20/20 to 20/400 (mean, 20/36; median, 20/30). Visual acuity at last follow-up ranged from 20/25 to counting fingers (mean, 20/56; median, 20/40). At last follow-up, 43 eyes (58%) had a visual acuity of 20/40 or better, and 57 (77%) had an improvement in visual acuity of 3 or more Snellen lines compared with their preoperative acuity. CONCLUSIONS: Macular hole closure and visual acuity improvement after initially successful macular hole surgery persist at follow-up of 5 years and longer in the majority of patients; delayed visual acuity improvement is not attributable to cataract surgery alone. PMID- 12719071 TI - Experimental study of viscoelastic in the prevention of corneal endothelial desiccation injury from vitreal fluid-air exchange. AB - PURPOSE: To evaluate the usefulness of viscoelastic in protecting the corneal endothelium from desiccation injury associated with fluid-air exchange in a rabbit model. DESIGN: Experimental study. METHODS: Rabbit eyes undergoing pars plana lensectomy and vitrectomy were insufflated with either dry or humidified air for 20 minutes following introduction of either Opegan (sodium hyaluronate 1.0%; Santen, Osaka, Japan) or Viscoat (sodium hyaluronate 3%-chondroitin sulfate 4%; Alcon, Tokyo, Japan) into the anterior chamber. In two other groups of rabbit eyes, the same procedure was performed without using any viscoelastic agent. Corneas obtained from rabbits undergoing surgery were compared with corneas obtained from rabbits not undergoing surgery. Potential alterations in the corneal endothelium were investigated by scanning electron microscopy, by Phalloidin-FITC staining of actin and by in vitro measurements of corneal permeability for carboxyfluorescein using a diffusion chamber. RESULTS: Scanning electron microscopy displayed less distortion of corneal endothelium with Opegan and Viscoat compared with the dry air-only exposed corneas. Using humidified air in Opegan and Viscoat coated corneas maintained the normal actin cytoskeleton during fluid-air exchange. Paracellular leakage was much less with Opegan and Viscoat use following infusion of dry air comparing to that of dry air-only group (P =.026 and P =.041). The difference was much more striking following humidified air infusion in Opegan or Viscoat coated corneas comparing to dry air-only infused corneas (P <.002 and P <.002). CONCLUSIONS: Coating of rabbit corneal endothelium with Opegan or Viscoat before fluid-air exchange largely prevents dry air damage to the endothelium. Infusion of humidified air further protects corneal endothelium during fluid-air exchange in aphakic rabbit eyes. PMID- 12719072 TI - Custom-designed plaque radiotherapy for nonresectable iris melanoma in 38 patients: tumor control and ocular complications. AB - PURPOSE: To evaluate plaque radiotherapy for iris melanoma. DESIGN: Prospective noncomparative interventional case series. METHODS: For 38 patients, custom designed plaque radiotherapy using iodine 125 isotope was applied overlying the cornea with a tumor apex dose of 80 Gy. The main outcome measures were tumor control and ocular complications using Kaplan-Meier estimates and Cox proportional hazards regression analysis. RESULTS: In all cases, the melanoma was nonresectable owing to large or discohesive tumor. The tumor configuration was nodular in 24 cases (63%) and flat (diffuse) in 14 (37%). The mean tumor basal diameter was 9 mm (range 4 to 13 mm). Solid tumor extended into the anterior chamber angle in 36 eyes (95%). Tumor seeds were noted on the iris stroma for a mean of 7 clock hours and in the anterior chamber angle for a mean of 4 clock hours. Five-year follow up revealed tumor metastasis in 0% and tumor recurrence in 8% of patients. Visual acuity of 20/200 or worse was found in 16% at 5 years. Radiation-related complications at 5 years included corneal epitheliopathy (9%), cataract (70%), and neovascular glaucoma (8%). No patients developed corneal necrosis, scleral necrosis, retinopathy, or papillopathy. After treatment, the combined incidence of tumor-related and radiation-related elevated intraocular pressure at 5 years was 33%. Enucleation was necessary in 13% at 5 years, for tumor recurrence (n = 3) and patient preference (n = 1). CONCLUSIONS: Plaque radiotherapy is a useful alternative to enucleation for eyes with nonresectable iris melanoma. Tumor control is 92% at 5 years, but related complications, especially cataract and elevated intraocular pressure, should be anticipated. PMID- 12719073 TI - Clinicopathologic findings in choroidal melanomas after failed transpupillary thermotherapy. AB - PURPOSE: To review the clinicopathologic features of eyes enucleated after failed transpupillary thermotherapy. DESIGN: Retrospective review. METHODS: Pathology reports in the L.F. Montgomery Laboratory between 1998 and 2002 were searched for enucleated eyes with choroidal melanoma that had been treated only by transpupillary thermotherapy (TTT) prior to enucleation. The clinical features of the patients, including ultrasonography examination, were reviewed and compared with the pathologic findings. RESULTS: Seven eyes from seven patients had been enucleated, representing 8% of eyes treated with TTT at our institute during the period studied. The primary indication for enucleation was tumor growth. The melanomas tended to grow laterally, with minimal if any increase in thickness. Five of the seven eyes histologically demonstrated extrascleral extension, which was detected by ultrasonography prior to enucleation in only one of those eyes. CONCLUSIONS: Choroidal melanoma may continue to grow along a path of least resistance after TTT. The melanoma may extend laterally in the choroid and through emissary canals. Early extrascleral extension may be difficult to detect by ultrasonography. PMID- 12719074 TI - Rate of progression of glaucoma correlates with retrobulbar circulation and intraocular pressure. AB - PURPOSE: To evaluate the correlation between progression rate of glaucomatous damage and retrobulbar blood flow in an institutional setting. DESIGN: Retrospective, observational case series. METHODS: Twenty open-angle glaucoma patients with at least five visual field examinations and progressive damage in at least one eye were included in the study. Mean +/- standard deviation follow up time was 4.3 +/- 1.6 years. As an indicator of progression rate of visual field damage, the angle to a horizontal line of the slope of the regression line of the visual field index mean defect over time was calculated for one randomly selected eye per patient. The association between this angle and intraocular pressure, as well as retrobulbar color Doppler imaging measurements obtained at the beginning of the observation period, was analyzed by a multiple linear regression analysis in a stepwise forward approach. RESULTS: With a faster rate in progression of glaucomatous damage, a lower baseline end diastolic blood flow velocity in the central retinal artery (partial r = -.63; P <.0037) and a higher baseline intraocular pressure (partial r =.59; P <.0078) were noted (multiple r =.69; P <.0043). Rate of progression was not related to the extent of preexisting visual field damage. CONCLUSIONS: Independent of the extent of glaucomatous damage and intraocular pressure, the progression rate of glaucomatous visual field damage statistically correlates with retrobulbar hemodynamic variables. PMID- 12719075 TI - Prevalence of organic neuro-ophthalmologic disease in patients with functional visual loss. AB - PURPOSE: To determine the concurrence of various patterns of functional visual loss (FVL) and organic disease. DESIGN: A retrospective case series in a university neuro-ophthalmology practice. METHODS: A retrospective case series in which we reviewed the clinical histories and neuro-ophthalmologic examinations of 133 consecutive patients diagnosed with FVL between July 1999 and August 2001. Functional visual loss was defined as an apparent afferent or efferent dysfunction that was unassociated with or far out of proportion with an identifiable lesion of the visual pathways. RESULTS: A total of 133 patients were identified. Nineteen were pediatric, and 76 were female. Fifty-six had concurrent head or eye pain. Nineteen had a recent history of trauma, and 17 had a recent surgical procedure. The most common pattern of FVL was a normal visual field in the presence of reduced visual acuity. Seventy-one patients (53%) with FVL presented with abnormal neuro-ophthalmologic examinations. Thirteen patients had central scotomata, and all had concurrent retinal or optic nerve pathology. Only 18 patients had a known disability claim pending. CONCLUSION: Although FVL was found in patients with no pathology, it was more common to find objective abnormalities on examination. This concurrence rate underscores the clinical necessity for following patients closely despite the diagnosis of FVL. No pattern of visual field constriction was routinely predictive of ophthalmologic or neurologic pathology except the presence of a central scotoma. The presence of a central scotoma in a nonorganic visual field should alert the practitioner to search for organic pathology. PMID- 12719076 TI - Visual loss associated with pediatric uveitis in english primary and referral centers. AB - PURPOSE: Pediatric uveitis is rare and has been reported to cause increased rates of visual loss compared with adult patients. The reasons for this are unclear. Only one study has been population-based, so the effect of referral bias is not known. We examined the pattern of disease in primary and referral centers to establish the unique characteristics of uveitis in children. DESIGN: Case control study. METHODS: Retrospective, multicenter, observational study of uveitis starting before the age of 20 years. Two hundred forty-nine patients were recruited from three primary and two referral ophthalmic units. Age-related differences in types of uveitis and systemic disease between hospitals were characterized, as were associations with visual loss. RESULTS: The incidence of uveitis in district hospitals at less than 16 years of age was 4.9/100,000: the most frequent diagnosis was idiopathic uveitis (78%). In referral cohorts the most frequent diagnosis was juvenile idiopathic arthritis-associated uveitis (67%). Other systemic diseases were rare. The most frequent type of uveitis at 0 to 7 years of age was chronic anterior uveitis, posterior uveitis in 8- to 15 year-olds, and acute anterior uveitis in 16- to 19-year-olds. Visual loss (any eye < 6/12) occurred in 17% and was not associated with age, sex, or hospital cohort. It was most frequent in posterior uveitis (25%). Treatment variables were independent predictors of visual loss: systemic treatment 2.2 (1.1- 4.6), surgical intervention 8.2 (3.8-17.6). CONCLUSIONS: Idiopathic uveitis was three times more common in district hospitals. Visual loss was similar to adult uveitis in this study. The increased frequency of severe chronic anterior uveitis in children aged 0 to 7 years and posterior uveitis in older children aged 8 to 15 years accounts for the rate of visual loss seen in previous studies. PMID- 12719077 TI - Spondylo-ocular syndrome: a new entity with crystalline lens malformation, cataract, retinal detachment, osteoporosis, and platyspondyly. AB - PURPOSE: To define a new clinical entity in a consanguineous family with six children affected by a spondylo-ocular syndrome, including cataract, crystalline lens malformation, retinal detachment, osteoporosis, and platyspondyly. To analyze candidate genes of connective tissue disorders as a possible underlying disorder and to demonstrate especially the ocular phenotype. DESIGN: Observational case series. METHODS: Consanguineous parents, one unaffected sibling and five affected children with clinical features of spondylo-ocular syndrome, were demonstrated. Clinical examination, radiologic, laboratory, and cytogenetic as well as moleculargenetic analyses were performed. The segregation of flanking marker alleles of three collagen genes and the interval for osteoporosis-pseudoglioma syndrome were analyzed. Two microsatellite markers located within Pax6CA/GT region were tested for homozygosity. RESULTS: On laboratory investigation a normal excretion of amino acids, mucopolysaccharides, and oligosaccharides could be found. The karyotype was normal. Complete radiologic examination in one index patient revealed a generalized moderate osteoporosis, platyspondyly with fish bone appearance, and greatly enlarged intervertebral spaces. The candidate genes known to be in Stickler syndrome as well as linkage to the osteoporosis-pseudoglioma syndrome candidate region could be excluded. None of the affected showed homozygosity for the Pax6 microsatellite markers. CONCLUSIONS: We conclude that the phenotype and the clinical features in this family defines a new Mendelian disorder. It remains to be seen what kind of molecule shared by eye and bone is involved. PMID- 12719078 TI - A comparison of latanoprost, bimatoprost, and travoprost in patients with elevated intraocular pressure: a 12-week, randomized, masked-evaluator multicenter study. AB - PURPOSE: To Internet Advance publication at ajo.com Feb 13, 2003. compare the intraocular pressure (IOP)-lowering effect and safety of latanoprost, bimatoprost, and travoprost in patients with open-angle glaucoma (OAG) or ocular hypertension (OH). DESIGN: Interventional study. METHODS: This 12-week, randomized, parallel-group study was conducted at 45 US sites. Previously treated patients with OAG or OH and an IOP > or =23 mm Hg in one or both eyes after washout received either latanoprost 0.005%, bimatoprost 0.03%, or travoprost 0.004% once daily in the evening. At baseline and after 6 and 12 weeks of therapy, masked evaluators measured IOP in triplicate at 8:00 AM, 12 noon, 4:00 PM, and 8:00 PM, and masked investigators graded conjunctival hyperemia before the 8:00 AM IOP measurement. The primary efficacy outcome measure was change between baseline and Week 12 in the 8:00 AM IOP (time of peak drug effect). RESULTS: In all, 410 of 411 randomized patients were included in intent-to-treat analyses (latanoprost, 136; bimatoprost, 136; travoprost, 138). Baseline mean 8:00 AM IOP levels were similar (P =.772); by week 12, reductions were observed in all 3 groups (P <.001 for each). Adjusted (ANCOVA) reductions in mean IOP at 8:00 AM were similar (P =.128) as were those at 12 noon, 4:00 PM, and 8:00 PM. Fewer latanoprost-treated patients reported ocular adverse events (P <.001, latanoprost vs bimatoprost), fewer reported hyperemia (P =.001, latanoprost vs bimatoprost), and average hyperemia scores were lower at week 12 (P =.001, latanoprost vs bimatoprost). CONCLUSIONS: Latanoprost, bimatoprost, and travoprost were comparable in their ability to reduce IOP in OAG and OH patients. Latanoprost exhibited greater ocular tolerability. PMID- 12719079 TI - Improved functional visual acuity after punctal occlusion in dry eye patients. AB - PURPOSE: To report an increased functional visual acuity, which was recently reported as a simulation of visual function of daily acts of gazing, in dry eye patients after punctal occlusion. DESIGN: Prospective comparative interventional study. METHODS: We measured ordinary best-corrected visual acuity and functional visual acuity in eight eyes of eight dry eye patients after punctal occlusion, and compared the results with those of 22 eyes of 22 dry eye patients without punctal occlusion. RESULTS: Functional visual acuity in dry eye patients after punctal occlusion was 0.962 in decimal notation, which was significantly higher than that of patients without punctal occlusion, 0.283 (P <.0001). CONCLUSIONS: This study shows that punctal occlusion can improve the impaired functional visual acuity of dry eye patients. PMID- 12719080 TI - Uveal effusion and angle-closure glaucoma in primary pulmonary hypertension. AB - PURPOSE: To report a patient with uveal effusion and intermittent angle-closure glaucoma associated with primary pulmonary hypertension. METHODS: Observational case report. RESULTS: A 78-year-old woman with primary pulmonary hypertension noticed pain and decreased vision in her right eye. Ocular examination disclosed excessive dilation of conjunctival and episcleral veins and uveal effusion with annular ciliochoroidal detachment in both eyes. The right eye revealed corneal edema with folds in the Descemet membrane, congested iris vessels, and a markedly shallow anterior chamber with a partially closed angle, corresponding to a recent attack of angle-closure glaucoma. Systemic treatment of her congestive heart failure with an angiotensin-II receptor antagonist resulted in a partial resolution of the uveal effusion and complete normalization of the anterior chamber depth. CONCLUSION: Primary pulmonary hypertension may cause uveal effusion, leading to a forward displacement of the lens-iris diaphragm and intermittent angle-closure glaucoma. PMID- 12719081 TI - The effect of optic disk edema on spontaneous venous pulsations. AB - PURPOSE: We evaluated the effect of optic disk edema on spontaneous venous pulsations (SVP) in patients without elevated intracranial pressure (ICP).Prospective, cross-sectional comparison in an institutional setting. METHODS: Twenty consecutive patients with unilateral optic disk edema due to anterior ischemic optic neuropathy (AION, n = 11) or optic neuritis (n = 9) were evaluated. The proportion of subjects with SVP in both eyes, the involved eye only, the uninvolved eye only, or neither eye was determined. Comparisons were made using the Exact McNemar Test. RESULTS: There was a significant difference in the proportion of uninvolved and involved eyes with SVP (60% [12/20] vs 5% [1/20], P <.005). Spontaneous venous pulsations were present in the uninvolved eye of 12 patients and the nonedematous portion of the optic disk in 1 patient with segmental edema from AION. CONCLUSIONS: Optic disk edema may cause SVP cessation without ICP elevation. PMID- 12719082 TI - American ophthalmology graduate medical education and the web: current state of internet resource utilization. AB - PURPOSE: To evaluate the presence and accessibility of ophthalmology graduate medical education (GME) programs on the internet and the degree to which web based resources are currently being utilized by American Ophthalmology training programs. DESIGN: Prospective sampling of internet postings of ophthalmology GME programs. METHODS: Ophthalmology residency programs listed in the American Medical Association Fellowship and Residency Electronic Interactive Database (FREIDA) online website (http://www.ama-assn.org/ama/pub/category/2997. html) were evaluated. Departmental websites were accessed either directly from the FREIDA website or from one of eight search engines or four metacrawlers utilized in the study. Each site was evaluated for the presence, extent, and type of postings in the following categories: residency, medical student resources, faculty, patient care, video utilization, and links. RESULTS: Departments with websites numbered 102 (83%). Specific posting rates were residency 84%; medical student resource 12%; faculty 72%; patient care 69%; video 2%; links 70%. CONCLUSIONS: Most departments have websites, but only a small number have postings in all categories. PMID- 12719083 TI - Unilateral leukocoria in off axis flash photographs of normal eyes. AB - PURPOSE: To describe an optical phenomenon producing leukocoria in off axis flash photographs of children who have a normal ophthalmic examination. DESIGN: Observational case series. METHODS: Description of three otherwise well children who presented to a pediatric ophthalmologist with unilateral leukocoria seen in flash photographs. The children underwent a full ophthalmic examination, including dilated fundus examination. RESULTS: All ophthalmic examinations were normal. The photographs were all found to be approximately 15 degrees off axis with the leukocoria seen in the eyes where the flash illuminated the nasal retina. CONCLUSIONS: Leukocoria can be seen in normal eyes when the child is not fixing directly on the camera. That is due to reflection off the optic nerve head. PMID- 12719084 TI - Camera artifacts producing the false impression of growth of choroidal melanocytic lesions. AB - PURPOSE: To report a camera artifact that masks portions of pigmented lesions while preserving overlying retinal details. DESIGN: Case report. METHODS: Cases collected from the authors' clinical practice were identified during review of fundus photos. An attempt was made to reproduce the artifact in a normal eye. RESULTS: Three cases were identified that demonstrated this artifact. CONCLUSION: Off-axis color photos can produce a reddish artifact that may blend imperceptibly with the surrounding retinal appearance and filter visualization of melanocytic pigment. This can mask a portion of a pigmented lesion and later may create a false impression of a larger lesion. PMID- 12719085 TI - Increased periocular pigmentation with ocular hypotensive lipid use in African Americans. AB - PURPOSE: To report increased eyelid pigmentation as an adverse side effect associated with topical ocular hypotensive lipids in African Americans. DESIGN: Interventional case series. METHODS: Two African-American patients with open angle glaucoma are described in whom increased eyelid pigmentation developed 1 month to 5 months after beginning treatment with either latanoprost or bimatoprost. RESULTS: Latanoprost was discontinued in an African-American patient, and pigmentation gradually diminished by 3 months after cessation of latanoprost. Increased eyelid pigmentation and increased eyelash length were noted in another African-American patient after just 4 weeks on bimatoprost. CONCLUSIONS: An increase in eyelid pigmentation and eyelash growth is a possible complication of topical ocular hypotensive lipid therapy, even in African American patients. The changes seems to present earlier after bimatoprost treatment then after latanoprost treatment. Cessation of these medications may lead to loss of induced pigmentation. PMID- 12719086 TI - A case of occult macular dystrophy accompanying normal-tension glaucoma. AB - PURPOSE: To present a case of occult macular dystrophy accompanying normal tension glaucoma. DESIGN: Observational case report. METHODS: Visual function and ophthalmoscopic and electroretinographic findings in a 56-year-old man suffering from gradually progressive photophobia and visual dysfunction were followed up over 4 years. RESULTS: Best-corrected visual acuity was below 20/200, and intraocular pressures remained in the low teens in both eyes. A wedge-shaped visual field defect in the right eye, corresponding to the enlarged disk cupping, and a relative central scotoma in the left were detected. Ophthalmoscopy and fluorescein angiography detected no apparent abnormality at the macular region in either eye. Severely attenuated multifocal electroretinographic responses were recorded from only the central areas, with normal full-field responses. CONCLUSION: Clinicians must look for additional pathologic conditions in patients with normal-tension glaucoma who have unexplained central visual dysfunction. PMID- 12719087 TI - A case of angle closure glaucoma caused by plateau iris and adie's pupil. AB - PURPOSE: To describe a case of plateau iris associated with bilateral Adie's pupil. DESIGN: Interventional case report. METHODS: A 54-year-old woman presented with pain in her right eye and headache. Intraocular pressure was 34 mm Hg in the right eye. Light reflex was defective bilaterally. RESULTS: Ultrasound biomicroscopic imaging revealed normal anterior chamber depth and narrow angle. The ciliary processes were situated anteriorly. The eyes showed supersensitivity to 0.125% pilocarpine. The patient was diagnosed as having bilateral angle closure glaucoma induced by Adie's pupil and plateau iris. CONCLUSION: Mild pupillary dilation caused by Adie's pupil may have played a role in the development of angle closure in the plateau iris configuration of our patient. PMID- 12719088 TI - Traumatic hyphemas in children secondary to corporal punishment with a belt. AB - PURPOSE: To report the severity of ocular injury in seven children with traumatic hyphemas resulting from the accidental striking of the child in the face with a belt during the administration of corporal punishment. DESIGN: Observational case series. METHODS: We retrospectively reviewed the records of all patients (n = 7, aged 4 to 14 years) with traumatic hyphemas secondary to belt injuries evaluated by the senior author between 1989 to 2002 at Le Bonheur Children's Medical Center, a regional pediatric referral hospital in Memphis, Tennessee. RESULTS: Anterior segment injuries ranged from small hyphemas with normal intraocular pressure and no vision loss to injuries with severe elevations of intraocular pressure and permanent, significant loss of vision. CONCLUSIONS: Ocular injury to a child can result from trauma inflicted with a belt by a parent or caretaker during corporal punishment and may result in permanent loss of vision. PMID- 12719089 TI - Myopic strabismus fixus: a mitochondrial myopathy? AB - PURPOSE: Ultrastructural analysis of extraocular muscle in a case of myopic strabismus fixus. DESIGN: Interventional case report. METHODS: Incisional biopsy and ultrastuctural analysis of recti muscles in a female patient with myopic strabismus fixus. RESULTS: Ultrastuctural analysis revealed evidence of mitochondrial myopathy. CONCLUSION: Acquired strabismus fixus due to high myopia may be a manifestation of mitochondrial myopathy, a finding that has not been previously reported. PMID- 12719090 TI - Severe proliferative retinopathy progressing to blindness in a japanese woman with takayasu disease. AB - PURPOSE: To report a patient with Takayasu disease with severe proliferative retinopathy progressing to bilateral blindness. A 57-year-old Japanese woman suffering from Takayasu disease for 16 years manifested severe proliferative retinopathy in both eyes, leading to blindness due to optic atrophy. DESIGN: Interventional case report. METHODS: A 57-year-old Japanese woman suffering from Takayasu disease for 16 years underwent a comprehensive opthalmologic examination. Panretinal photocoagulation and vitrectomy were performed on both eyes. SETTINGS: Institutional practice. RESULTS: At initial examination, the patient was found to have severe proliferative retinopathy with tractional retinal detachment in both eyes. The retinal detachments were repaired by the vitrectomy, however, the patient developed bilateral blindness due to optic atrophy. CONCLUSIONS: Patients with a long history of Takayasu disease can demonstrate severe proliferative retinopathy that may be resistant to photocoagulation and/or vitreous surgery. Ocular ischemia can lead to blindness from ischemic optic neuropathy. PMID- 12719091 TI - Leber congenital amaurosis associated with optic disk neovascularization and vitreous hemorrhage. AB - PURPOSE: To report an unusual case of optic disk neovascularization and vitreous hemorrhage associated with Leber congenital amaurosis (LCA). DESIGN: Interventional case report. METHODS: A 16-year-old Caucasian girl with a history of LCA presented with decreased vision in her left eye, diffuse retinal pigmentary abnormalities characteristic of LCA, and hemorrhage over the left optic disk and macula. Six months of follow-up revealed optic disk neovascularization. A small amount of neovascularization was noted in the right eye at 6 months. RESULTS: An extensive systemic evaluation indicated no other cause for the neovascularization. Panretinal photocoagulation was performed in both eyes, and subsequently the neovascularization regressed. CONCLUSIONS: Leber congenital amaurosis like retinitis pigmentosa, can rarely be associated with neovascularization of the disk, which is amenable to treatment with peripheral photocoagulation if it does not spontaneously regress. PMID- 12719092 TI - Foveal thickness in occult macular dystrophy. AB - PURPOSE: Occult macular dystrophy (OMD) is an inherited macular dystrophy characterized by a progressive macular dysfunction without any visible fundus abnormality. We studied the foveal thickness in patients with OMD using optical coherence tomography (OCT). DESIGN: Observational case series. Foveal thickness by OCT images. METHODS: Foveal thickness obtained from 22 eyes of 11 patients with OMD was compared with that from 27 eyes of 20 age-matched normal controls. RESULTS: Mean foveal thickness in the patients group (96.5 +/- 19.5 microm) was significantly thinner than that in the normal controls (133.3 +/- 9.0 microm, P <.0001, Mann-Whitney U test). Eighteen of 22 eyes with OMD had foveal thickness that were thinner than the lower limit of the normal range. There was no statistically significant correlation between the foveal thickness and visual acuity, age, or duration from onset. CONCLUSION: These results demonstrated that there are significant anatomic changes in the macula of patients with OMD. PMID- 12719093 TI - Consecutive choroidal melanoma in the same eye of a patient. AB - PURPOSE: To report the occurrence of two entirely separate choroidal melanomas in the same eye of one patient. DESIGN: Observational case report. METHODS: Retrospective case report from one clinical practice. Two choroidal melanomas were observed to develop in the same eye of one patient. RESULTS: An inferonasal choroidal melanoma was diagnosed in the left eye of a 30-year-old woman in 1990. This was successfully treated with proton beam radiotherapy. In 1996, a superotemporal choroidal tumor in the same eye was observed to grow rapidly and it was treated with a ruthenium plaque in 1996. There was a history of cutaneous malignant melanoma. CONCLUSIONS: It is extremely rare for two choroidal melanomas to develop in one eye, and there may be a genetic predisposition. PMID- 12719094 TI - Retinal tear and detachment after transpupillary thermotherapy for choroidal melanoma. AB - PURPOSE: To report a side effect of transpupillary thermotherapy for choroidal melanoma. DESIGN: Small case series. METHODS: Review of clinical charts and photographs in a private retina practice. RESULTS: Retinal tears developed in two patients after transpupillary thermotherapy for choroidal melanoma. In one patient a secondary rhegmatogenous retinal detachment developed. CONCLUSIONS: Transpupillary thermotherapy can result in vitreous traction on the retina over the choroidal tumor, which can lead to a retinal tear and to retinal detachment. Patients should be warned about this possibility and advised to be examined should photopsias or new floaters develop after treatment. PMID- 12719095 TI - Congenital iris coloboma repair using a modified McCannel suture technique. AB - PURPOSE: To report a repair of congenital iris coloboma using a modified McCannel suture technique. DESIGN: Interventional case report. METHODS: Retrospective intervention procedure. RESULTS: Postoperative repair resulted in restoration of pupil, anterior support of posterior chamber intraocular lens, and improved appearance. CONCLUSIONS: The modified McCannel suture technique provides acceptable functional and cosmetic repair of congenital iris colobomas. PMID- 12719096 TI - Visual outcomes and incidence of vitreous loss for residents performing phacoemulsification without prior planned extracapsular cataract extraction experience. AB - PURPOSE: Phacoemulsification cataract surgery is one of the most important surgical procedures learned by ophthalmology residents during their residency training. We evaluated the visual outcomes and incidence of vitreous loss of phacoemulsification cataract surgeries performed by ophthalmology residents without prior planned extracapsular cataract extraction experience. DESIGN: Interventional case series. METHODS: A retrospective review was performed on 332 consecutive phacoemulsification cataract surgeries performed by third-year ophthalmology residents from July 1999 through June 2001. Data included are preoperative and postoperative best-corrected visual acuity, preexisting ocular comorbidities, and intraoperative and postoperative complications. RESULTS: Postoperative best-corrected visual acuity was 20/40 or better in 89% of eyes. After excluding the cases with preexisting ocular comorbidities, the percentage increased to 97.7%. Vitreous loss occurred in 4.8% of cases. CONCLUSIONS: Ophthalmology residents can learn to perform phacoemulsification cataract surgery safely and effectively without prior planned extracapsular cataract extraction experience. PMID- 12719098 TI - How to write a brief report for the AJO. PMID- 12719097 TI - A novel CACNA1F mutation in a french family with the incomplete type of X-linked congenital stationary night blindness. AB - PURPOSE: To describe a French family with the incomplete type of X-linked congenital stationary night blindness (CSNB2) associated with a novel mutation in the retina-specific calcium channel alpha(1) subunit gene (CACNA1F). DESIGN: Interventional case report. METHODS: Two family members with a history of nonprogressive night blindness and subnormal visual acuity were clinically examined and the genotype determined by molecular genetic analysis. RESULT: Both patients had clinical manifestations characteristic of CSNB2. Electrophysiologically, we found a predominant reduction of the ERG B-wave in the maximal response. Both rod and cone function were subnormal, with the latter tending to be more attenuated. We identified a C deletion at nucleotide position 4548, resulting in a frameshift with a predicted premature termination at codon 1524. CONCLUSIONS: The clinical and genetic study of a novel mutation in the CACNA1F gene adds further support to the contention that CSNB2 represents a genetically distinct retinal disorder of a calcium channel. PMID- 12719099 TI - Treatment of anemia in the diabetic patient with retinopathy and kidney disease. PMID- 12719100 TI - Optical coherence tomography measurement of nerve fiber layer thickness and the likelihood of a visual field defect. PMID- 12719103 TI - Correlation between retinal abnormalities and intracranial abnormalities in the shaken baby syndrome. PMID- 12719104 TI - Keratocyte density in keratoconus. PMID- 12719106 TI - Bilateral presumed endogenous candida endophthalmitis and stage 3 retinopathy of prematurity. PMID- 12719108 TI - Macular translocation with 360-degree retinotomy for management of age-related macular degeneration with subfoveal choroidal neovascularization. PMID- 12719109 TI - Validity of a new disk grading scale for estimating glaucomatous damage: correlation with visual field damage. PMID- 12719125 TI - Conformational changes of pediocin in an aqueous medium monitored by fourier transform infrared spectroscopy: a biological implication. AB - Fourier transform infrared (FTIR) spectroscopy was used to investigate the secondary structure of pediocin PA-1 in different aqueous media in relation to its antimicrobial activity. The experiments were performed at pD (pH meter corrected for deuterium isotope effect) 6, 7, and 8 and during a heating-cooling cycle of 20-80 degrees C. At pD 6, (i.e. pediocin's most active form), the FTIR results show that pediocin adopts an unordered structure with a small contribution of beta-turn. After a heating-cooling cycle, thermally-induced changes in pediocin are reversed and its activity is maintained. Increasing the pD to 7 and 8 leads to a more ordered secondary structure. For these two pD values, an increase in temperature induces an irreversible aggregation of protein as revealed by the amide I' band. The analysis of the Tyr region provides more insight into the aggregation process. In fact, it appears to be a two-step process, involving first the C (carboxy)-terminus of pediocin and then the N (amino)-terminus. This study reveals two major points: (1) the preservation of pediocin flexibility is essential for maintaining its activity; and (2) the aggregation of its C-terminus is sufficient to induce a loss of activity, suggesting that this region plays an important role in the activity of pediocin. PMID- 12719126 TI - Disaggregating effects of ethanol at low concentration on beta-poly-L-lysines. AB - Protein aggregation is involved in a number of disorders, such as Alzheimer's disease, cystic fibrosis, and prion diseases. Such aggregates are formed by peptides in beta-conformation. The study of the processes of aggregation or its inhibition makes it necessary for the peptide to remain in a monomeric state at the beginning of aggregation assays. Using three poly-L-lysine as a model of beta peptide, we measured the spectral changes occurring in the visible spectrum of Congo Red (CR), a diazo dye, in two solvent media, namely, an aqueous solution of ethanol 10% (v/v), and an aqueous solution of dimethyl sulfoxide (DMSO) 5% (v/v). Aggregation constants show that the presence of ethanol at low concentration produces a disaggregating effect, regardless of the degree of polymerisation of the peptide. This effect is considered to be due to the direct binding of ethanol molecules to the peptide. This binding undergoes an enhancement of the electrostatic repulsion among charged lysine chains. PMID- 12719127 TI - Poly(ethylene glycol)-grafted poly(3-hydroxyundecenoate) networks for enhanced blood compatibility. AB - Poly(ethylene glycol)-grafted poly(3-hydroxyundecenoate) (PEG-g-PHU) networks were prepared by irradiating homogeneous solutions of poly(3-hydroxyundecenoate) (PHU) and the monoacrylate of poly(ethylene glycol) (PEG) with UV light. The resulting polymer networks were characterized by measuring the water contact angle, water uptake, and mechanical properties and by performing attenuated total reflectance infrared spectroscopy and scanning electron microscopy. These measurements showed that the PEG chains were present in polymer networks. Adsorption of blood proteins and platelets on cross-linked PHU (CLPHU) and PEG-g PHU were examined using poly(L-lactide) (PLLA) surfaces as control. Blood proteins and platelets had significantly lower tendency of adhesion to surfaces composed of CLPHU and PEG-g-PHU networks than to PLLA. Blood compatibility of polymer networks increased as the fraction of grafted PEG increased. The results of this study suggest that PEG-g-PHU networks might be useful for blood compatible biomedical applications. PMID- 12719128 TI - Synthesis of chitooligosaccharide derivative with quaternary ammonium group and its antimicrobial activity against Streptococcus mutans. AB - A derivative of chitooligosaccharide (COS) with quaternary ammonium functionality was synthesized and characterized by means of FT-IR and NMR spectroscopy. Its amtimicrobial activity was evaluated against Streptococcus mutans, which is a principal etiological agent of dental caries in humans. Introduction of quaternary ammonium group to COS has been easily accomplished by coupling of glycidyl trimethylammonium chloride (GTMAC) to COS in aqueous solution without an additional catalyst. The degree of substitution (%), as determined by (1)H NMR, of GTMAC to the COS increased up to 116% at 70 degrees C for 24h. The resulting COS-GTMAC exhibited the growth inhibition of above 80% against S. mutans after 5h, whereas the COS showed the growth inhibition of about 10%. It was found that antimicrobial activity of the COS could be considerably enhanced by the introduction of quaternary ammonium functionality. PMID- 12719129 TI - Tensile deformation of bacterial cellulose composites. AB - The polymeric basis for the mechanical properties of primary plant cell walls has been investigated by forming analogous composites based on fermentation of the bacterium Acetobacter xylinus, either alone or in the presence of xyloglucan or pectin. Simultaneous small-angle X-ray scattering and uniaxial deformation experiments has shown how the cellulose microfibrils reorient during deformation. Despite very different stress/strain curves, the reorientation behaviour is similar, regardless of the presence or absence of xyloglucan or pectin. A simple theory has been developed to predict the orientation behaviour. This is qualitatively similar to the measured behaviour, but differs quantitatively. PMID- 12719130 TI - Preparation of self-assembled silk sericin nanoparticles. AB - Silk sericin (SS) possessing moisture-retaining property was reacted with activated poly(ethylene glycol) (PEG) to obtain self-assembled SS nanoparticles. The aliphatic and aromatic hydroxyl groups of serine and tyrosine residues as the reaction sites in SS were clarified by amino acid analysis and 1H NMR spectroscopy, respectively. From IR and circular dichroism (CD) measurements, introduction of PEG into SS induced the conformational change from random coil to beta-sheet. DSC thermogram of sericin-PEG conjugate suggests that mutual miscibility between PEG and SS chains was poor. Nanoparticles of sericin-PEG conjugate with sizes measured by dynamic light scattering ranging about 200-400 nm in diameter, were prepared by the diafiltration method. Shape of sericin-PEG conjugate nanoparticles observed by scanning and transmission electron microscopes was spherical. The results suggest that sericin-PEG conjugates are self-associated to form spherical nanoparticles through hydrophobic interaction. PMID- 12719131 TI - Equilibrium unfolding of dimeric human prostatic acid phosphatase involves an inactive monomeric intermediate. AB - Guanidine hydrochloride (GdnHCl)-induced unfolding of human prostatic acid phosphatase (hPAP), a homodimer of 50 kDa subunit molecular weight, was investigated with activity measurements, size exclusion HPLC, tryptophan fluorescence, 1-anilinonaphtalene-8-sulfonate (ANS) binding and reactivity with 2 (4'-maleimidoanilino)naphthalene-6-sulfonate (MIANS). Equilibrium analysis was performed to shed light on the role of dimerization in the folding and stability of the catalytically active oligomeric protein. Unfolding was reversible, as verified by activity measurements and tryptophan fluorescence. The noncoincidence of the unfolding curves obtained by different techniques suggests the occurrence of a multiphasic process. The reaction of hPAP inactivation is accompanied by dissociation of the dimer into two monomers. The midpoint of this transition is at 0.65 M GdnHCl with 4.24+/-0.12 kcalmol(-1) free energy change. Binding of ANS to the inactive phosphatase monomer, especially remarkable in the region from 0.8 to 1.25M GdnHCl, suggests that the hydrophobic probe indicates exposition of the intersubunit hydrophobic surface and a loosening of the monomer's tertiary structure. Strong fluorescence of thiol group derivatives, the products of their reaction with MIANS, appears in a limited range of GdnHCl concentrations (1.2 1.6M). This shows that in the relaxed structure of the intermediate, the reagent is allowed to penetrate into the hydrophobic environment of the partially hidden thiol groups. The equilibrium unfolding reaction of hPAP, as monitored by tryptophan fluorescence, does not depend on the protein concentration and displays a single transition curve with a midpoint at 1.7 M GdnHCl and value of DeltaG(unf)(H(2)O)=3.38+/-0.08 kcalmol(-1) per monomer, a result implying that this transition is related to the conformational change of the earlier dissociated and already inactive subunit of the protein. PMID- 12719132 TI - Seroepidemiology of Toxoplasma gondii in dogs in Trinidad and Tobago. AB - A cross-sectional study was conducted to determine the seroprevalence of Toxoplasma gondii agglutinins and to investigate the relationship between various risk factors and occurrence of toxoplasmosis in dogs in Trinidad. Of a total 250 dogs, comprising domestic, hunting and stray dogs, 80 (32.0%) were positive for T. gondii agglutinins at a titre of > or =1:32 using a latex agglutination test. Stray dogs (60.5%) had statistically significantly higher (P<0.001) seroprevalence for toxoplasmosis than hunting dogs (30.5%) and domestic dogs (25.5%). Amongst dogs whose ages were known, the prevalence of toxoplasmosis was significantly highest (P=0.037) in dogs in the >2-3 years age group compared with other age groups. Dogs that consumed home-cooked foods had a seroprevalence of 32.9% compared with those fed commercial dog foods (17.2%) and dogs fed both home cooked and commercial foods (21.0%). However, the difference was not statistically significant (P>0.05; chi(2)). The rather high prevalence of toxoplasmosis in stray dogs is a good indication of the extent of the infection in the environment. PMID- 12719133 TI - Molecular studies on Babesia, Theileria and Hepatozoon in southern Europe. Part I. Epizootiological aspects. AB - Molecular epizootiology of piroplasmids (Babesia spp., Theileria spp.) and Hepatozoon canis was studied in mammals from southern Europe (mainly from Spain, but also from Portugal and France). Partial amplification and sequencing of the 18s rRNA gene was used for molecular diagnosis. In some particular cases (B. ovis and B. bovis) the complete 18s rRNA gene was sequenced. Blood samples were taken from domestic animals showing clinical symptoms: 10 dogs, 10 horses, 10 cows, 9 sheep and 1 goat. In addition, DNA samples were isolated from blood of 12 healthy dogs and from spleen of 10 wild red foxes (Vulpes vulpes). The results of the survey were the following: Piroplasmid infections: Approximately from 50 to 70% of wild or domestic mammals (symptomatic) were infected. Piroplasmids detected in ruminants were:COW: B. bovis, T. annulata and Theileria sp. (type C). Sheep and goat: B. ovis. Piroplasmids present in canids were: Babesia canis vogeli, Babesia canis canis, Theileria annae and B. equi. The only piroplasmid found in asymptomatic dogs was B. equi. Piroplasmids found in horse were: B. equi and B. canis canis.H. canis infections in canids: H. canis was absent of domestic dog samples, whereas all foxes studied were infected by this protozoa. Genetic analysis showed that most of piroplasmid and Hepatozoon isolates from southern Europe matched unambigously with previously described species, as demonstrated by the high level sequence identity between them, usually between 99 and 100%. Minor differences, usually detected in hypervariable regions of 18s rRNA gene are probably due to strain variations or rare genetic polymorphisms. A possible exception was B. bovis, which shows a relatively lower degree of homology (94%) with regard to other B. bovis isolates from several countries. The same is true for B. ovis, that showed a 94% identity with regard to Babesia sp. from South African cow and a 92% with rapport to B. bovis from Portugal. PMID- 12719134 TI - Humoral immune response in adult blue foxes (Alopex lagopus) after oral infection with Encephalitozoon cuniculi spores. AB - Encephalitozoon cuniculi causes severe diseases in blue fox puppies. When pregnant vixens are infected, parasites are transmitted over the placenta to the unborn that subsequently develop encephalitozoonosis. Adult foxes themselves do not have signs of disease, but show antibody titres to E. cuniculi. The purpose of the present study was to gain information on the immune response in adult foxes after experimental infection. Sixteen foxes were infected orally with E. cuniculi spores, eight of them twice and 28 days apart. The two groups of animals showed elevated serological values in both the carbon immunoassay and in the ELISA. Elevated serological levels were recorded up to 1 year after the infection took place. The control group (n=8) remained serologically negative throughout the trial. The results of the study showed that blue foxes could be seropositive for at least a year after oral infection with E. cuniculi. PMID- 12719135 TI - A rapid MTT colorimetric assay to assess the proliferative index of two Indian strains of Theileria annulata. AB - A study was undertaken to compare the proliferative index of macroschizont infected lymphoblastoid cells of two Indian strains [Izatnagar (IZT) and Parbhani (PBN)] of Theileria annulata by an in vitro MTT [3-(4,5-dimethyl thiazol-2-yl) 2,5-diphenyl tetrazolium bromide], colorimetric assay. Culture conditions were standardized to define the optimal cell concentration in 96-well microculture plates to yield nearly 100% living cells for measurement of the metabolized formazan activity. A cell concentration of 1.5x10(5) cells/ml was found to be optimal for effective discrimination of the parasite strains. On the basis of conversion of MTT by the actively proliferating lymphoblastoid cells, the PBN strain of T. annulata stimulated a 2.5-fold increase in formazan activity in comparison to the IZT strain. The in vitro MTT assay was found to be a simple and convenient method for assessing the cell activation rate and growth, obviating the need for radioactive material for the assay. The results of the proliferation assay are discussed in relation to previously documented information on the biological characteristics of this important pathogen of cattle. PMID- 12719136 TI - Characterization of the protective response against a homologous challenge infection with Strongyloides venezuelensis in rats. AB - The protective response in rats against a homologous challenge infection with Strongyloides venezuelensis was characterized. In an initial infection with 1000 filariform larvae and migrating larvae (L(3)) of S. venezuelensis, the population of L(3) in the lungs on day 3 postinfection (PI), and that of adult worms in the small intestine on day 7 PI, were 180.8+/-14.5 and 336.8+/-70.7, respectively. The latter were gradually expelled towards day 42 PI. After the initial infection, the rats developed strong immunity against a homologous challenge infection as manifested by a marked reduction in worm populations, stunted body length and width, damage to reproductive organs, impaired egg production and rapid expulsion of the worms by day 14 after challenge. Expulsion of the worms was preceded by a significantly elevated (P<0.05) peripheral blood eosinophil (PBE) count, both in the initial (200.0+/-26.5 x 10(3)ml) and the challenge infection (400.9+/-165.4 x 10(3)ml). These findings suggest that rats acquire strong homologous immunity following initial exposure to S. venezuelensis. It is suggested that PBEs are involved in worm expulsion. A major target of these effector mechanisms is the reproductive system of S. venezuelensis. PMID- 12719137 TI - Cerebellar ataxia due to Toxocara infection in Mongolian gerbils, Meriones unguiculatus. AB - We assessed the usefulness of gerbils as an experimental model for neurologic toxocarosis. Mongolian gerbils, Meriones unguiculatus, infected with Toxocara canis or Toxocara cati (1000 eggs/gerbil) showed progressive neurologic disorders from 50 days after infection in T. canis-infected gerbils or from 120 days after infection in T. cati-infected gerbils. The incidence of the onset was 6 of the 13 gerbils (49%) in the T. canis-gerbils and 5 of the 7 gerbils (71%) in the T. cati gerbils. Histopathologically, the cerebellum was the most affected in both groups. We observed loss of Purkinje cells, glial nerve fibers, and nerve sheaths. We also found foci consisting of aggregated macrophages scattered in the white matter of the cerebellum. The affected gerbils showed ataxia and ultimately died of cachexia. Our findings suggest that irreversible neurologic toxocarosis in gerbils can be induced by infection with either T. canis or T. cati. PMID- 12719138 TI - Canine dirofilariasis in the region of Conceicao Lagoon, Florianopolis, and in the Military Police kennel, Sao Jose, State of Santa Catarina, Brazil. AB - Eighty dogs from a locality west of the Conceicao Lagoon, in the Santa Catarina Island, Florianopolis Municipality, State of Santa Catarina, Brazil, were examined for dirofilariasis by thick smear and by modified Knott's method. Twelve of them (15%) were positive, five only by Knott's, two only by thick smear and five by both methods. Positive dogs were observed in several parts of the locality, indicating that the parasite is widespread. Prevalence was significantly higher in older animals. Size, sex, hair length, place of sleeping and hour of blood collection did not significantly influence the results. Forty dogs of the Military Police kennel, at Sao Jose Municipality, Santa Catarina State, were all negative. Studies on the transmission and control of dirofilariasis in the region should be developed. PMID- 12719139 TI - An estimation of Toxocara canis prevalence in dogs, environmental egg contamination and risk of human infection in the Marche region of Italy. AB - The human risk of infection with larvae of Toxocara canis was estimated in people from the Marche region of Italy. This region includes both urban and rural areas and its inhabitants frequently keep dogs for company, hunting, as guardians or shepherds. T. canis infection was diagnosed in 33.6% out of 295 dogs examined. Nearly half of the dogs (48.4%) living in rural areas were found T. canis positive, compared to about one-quarter of the dogs (26.2%) from urban areas. Analysis by provenance and role revealed the highest infection rate in rural hunting dogs (64.7%) and the lowest in urban companion dogs (22.1%). According to questionnaire data, the peridomestic environment, i.e. gardens and dog pens, is the most important defecation site in both rural and urban areas. Since over 40% of the dogs who defecate in dog pens are infected and 24% of urban and 47% of rural dogs who leave their droppings in the house surroundings harbour the parasite, it is clear that these environments may constitute sites of zoonotic risk. Our analysis of soil samples from 60 farms confirmed the high contamination level, revealing positive soil samples in more than half of the farms. Substantial egg contamination was also found in urban areas, as 3/6 parks examined were Toxocara spp. positive. Finally, our serological findings indicate that human infection actually occurs in the area: 7 out of 428 adults examined (1.6%) had very high levels of antibodies to T. canis antigen, suggesting a previous contact with the Larva migrans of the nematode. PMID- 12719140 TI - Effects of condensed tannins on goats experimentally infected with Haemonchus contortus. AB - Although the use of tanniferous plants or condensed tannins as an alternative to anthelmintics to control gastrointestinal nematodes has been largely documented in sheep, studies remain scarce in goats. The objective of this study was therefore to assess the possible impact of condensed tannins in goats infected with adult Haemonchus contortus. Two groups of cull goats were experimentally infected with 10.000 L3 of H. contortus. After 4 weeks, quebracho extracts, representing 5% of the diet DM, were administered for 8 days to one of the two groups. Goats of the second group remained as controls. One week after the end of quebracho administration, the goats were euthanised. Individual egg excretion and pathophysiological parameters were measured weekly during the study. At the end of the study, worm counts were assessed and histological samples from the abomasa were taken to count the numbers of mucosal mast cells, globule leukocytes and eosinophils. The administration of tannins was associated with a significant decrease in egg excretion, which persisted until the end of experiment. This reduction was not associated with any difference in worm number but with a significant decrease in female fecundity. No significant changes in the mucosal density of the three inflammatory cell types were detected between the two groups. These results indicate that the major consequence of tannin consumption in goats is a reduction in worm fecundity and egg output, which does not seem related to significant changes in the local mucosal response. PMID- 12719141 TI - Effect of treatment on the dynamics of circulating hypodermin C in cattle naturally infested with Hypoderma lineatum (Diptera: Oestridae). AB - An antigen capture ELISA, using a murine monoclonal antibody recognising recombinant hypodermin C (rHyC), was used to evaluate the influence of early treatment with eprinomectin (Eprinex) or fenthion (Spotton) on the kinetics of circulating hypodermin C in calves naturally infested with Hypoderma lineatum. No viable larvae were collected from treated animals, whereas a variable number of warbles were found in control animals. Treatment provoked a decrease in circulating HyC levels that was significant 9 days post-treatment (p.t.). Circulating antigen levels in the treated cattle remained detectable for approximately 99 days p.t. In contrast, control animals had no detectable antigen at 64 days p.t., 42 days earlier than in the treated animals. These results suggest that larvae were either gradually killed, resulting in slow release of antigen or they were encapsulated, leading to the slow liberation of antigen. Kinetics of circulating HyC did not differ among the two insecticide treatments. Antibodies persisted, in all groups, throughout the 120-day study. These results suggest that the antigen capture ELISA will be useful as a technique for detecting successful treatment of cattle grub infestations and for the detection of new infestations in previously infested cattle. PMID- 12719143 TI - Exotic ticks introduced into the United States on imported reptiles from 1962 to 2001 and their potential roles in international dissemination of diseases. AB - Since 1962, a total of 29 species of exotic ticks have been introduced into the United States on imported reptiles, with 17 species from the genus Amblyomma, 11 from the genus Aponomma and one from the genus Hyalomma. In the absence of measures to control introduction of these importations, some exotic tick species will develop breeding colonies and become established as indigenous species and some tickborne diseases may be introduced to wreak havoc among susceptible native populations. However, formulation of risk assessments and rational control measures have been hampered by a lack of knowledge of these exotic ticks, with much of the available data published in older and relatively obscure publications. This report is an attempt to collate information for all 29 exotic tick species, including previously unpublished data from our laboratory, with particular reference to their geographical distribution, hosts, life cycles and vector potential, and to review methods to minimize their global dissemination. PMID- 12719142 TI - Amblyomma aureolatum (Pallas, 1772) and Amblyomma ovale Koch, 1844 (Acari: Ixodidae): hosts, distribution and 16S rDNA sequences. AB - DNA sequences of Amblyomma aureolatum (Pallas, 1772) and Amblyomma ovale Koch, 1844 were obtained to determine genetic differences between these tick species. Collections of these species are discussed in relation to distribution and hosts. Seven ticks collections (four from Brazil, one from Argentina, one from Uruguay and one from USA) house a total of 1272 A. aureolatum (224 males, 251 females, 223 nymphs and 574 larvae) and 1164 A. ovale (535 males, 556 females, 66 nymphs and 7 larvae). The length of the sequenced mitochondrial 16S rRNA gene fragment for A. aureolatum was 370bp and for A. ovale was 373bp. The DNA sequence analysis showed a 13.1% difference between the two species. Apart from one male A. ovale found on a toad, all adult ticks were found on mammals. The majority of adult specimens of both tick species were removed from Carnivora (96.1 and 84.3% of A. aureolatum and A. ovale, respectively), especially from dogs (53.1% of A. aureolatum, and 46.4% of A. ovale). Collections on wild Canidae were higher for A. aureolatum (23.3%) than for A. ovale (7.1%). On the other hand, collections of A. ovale adults on wild Felidae were higher (18.3%) than findings of A. aureolatum (9.2%). The contribution of other mammalian orders as hosts for adults of A. aureolatum and A. ovale was irrelevant, with the exception of Perissodactyla because Tapiridae contributed with 13.0% of the total number of A. ovale adults. Adults of both tick species have been found occasionally on domestic hosts (apart of the dog) and humans. Most immature stages of A. aureolatum were found on Passeriformes birds, while rodents and carnivores were the most common hosts for nymphs and larvae of A. ovale. A. aureolatum has been found restricted to the Neotropical region, covering the eastern area of South America from Uruguay to Surinam, including northeastern Argentina, eastern Paraguay, southeastern Brazil and French Guiana. A. ovale showed a distribution that covers the Neotropical region from central-northern Argentina throughout the Neotropics into the Nearctic region of Mexico with a few records from the USA, also with collection sites in Paraguay, Bolivia, most Brazilian states, Peru, Ecuador, French Guiana, Surinam, Guyana, Trinidad & Tobago, Venezuela, Colombia, Panama, Costa Rica, Nicaragua, Belize, Guatemala and several states of Mexico. PMID- 12719144 TI - Molecular characterization of Blastocystis isolates from primates. AB - Twelve Blastocystis isolates from primates were analyzed genetically by polymerase chain reaction (PCR) using diagnostic primers and PCR-restriction fragment length polymorphism (RFLP) of SSUrDNA. Two distinct genotypes, subtype 1 and a variant of subtype 1, were detected in two and six of the 12 isolates, respectively. The RFLP profiles of the isolates designated as subtype 1 were identical to the profile of ribodeme 1. The RFLP profiles of the six isolates designated as variants of subtype 1 were different from the profile of the variant of subtype 1 from a human reported previously. The other four isolates were not amplified with any diagnostic primers, but three of them showed the same RFLP profiles as ribodeme 6. This study was the first genomic analysis of Blastocystis isolates from primates, and showed the genetic similarity between the isolates from primates and the genotypes of Blastocystis hominis. However, it was unclear whether the isolates examined were zoonotic or not. Therefore, it is necessary to reveal the phylogenetic relationships between the isolates from primates and the multiple genotypes of B. hominis. PMID- 12719145 TI - Increase of Th1 type cytokine mRNA expression in peripheral blood lymphocytes of calves experimentally infected with Cryptosporidium parvum. AB - The expression of the messenger RNA of interleukin-12 (IL-12), interferon-gamma, interleukin-4, interleukin-6, and interleukin-10 was examined by reverse transcriptase polymerase chain reaction in peripheral blood lymphocytes of calves that were orally inoculated with Cryptosporidium parvum oocysts. In all of the calves, gene expression of interleukin-12, interleukin-6, and interferon-gamma was observed at delivery and this expression was repressed within the next 24h. In calves inoculated with C. parvum, mRNA expression of interleukin-12 and interferon-gamma was noticed on day 3 post-inoculation (p.i.) and increased in the convalescent phase of the infection, whereas in non-inoculated calves no mRNA expression was detectable up to the end of the experiment. No mRNA expression of interleukin-4 or 6 was detected during the experiment. Our observations suggest that systemic Th1 type immune responses are induced in calves infected with C. parvum and may be available for evaluation of the control of the infection. PMID- 12719147 TI - DISPLAY-2: a two-dimensional shallow layer model for dense gas dispersion including complex features. AB - A two-dimensional shallow layer model has been developed to predict dense gas dispersion, under realistic conditions, including complex features such as two phase releases, obstacles and inclined ground. The model attempts to predict the time and space evolution of the cloud formed after a release of a two-phase pollutant into the atmosphere. The air-pollutant mixture is assumed ideal. The cloud evolution is described mathematically through the Cartesian, two dimensional, shallow layer conservation equations for mixture mass, mixture momentum in two horizontal directions, total pollutant mass fraction (vapor and liquid) and mixture internal energy. Liquid mass fraction is obtained assuming phase equilibrium. Account is taken in the conservation equations for liquid slip and eventual liquid rainout through the ground. Entrainment of ambient air is modeled via an entrainment velocity model, which takes into account the effects of ground friction, ground heat transfer and relative motion between cloud and surrounding atmosphere. The model additionally accounts for thin obstacles effects in three ways. First a stepwise description of the obstacle is generated, following the grid cell faces, taking into account the corresponding area blockage. Then obstacle drag on the passing cloud is modeled by adding flow resistance terms in the momentum equations. Finally the effect of extra vorticity generation and entrainment enhancement behind obstacles is modeled by adding locally into the entrainment formula without obstacles, a characteristic velocity scale defined from the obstacle pressure drop and the local cloud height.The present model predictions have been compared against theoretical results for constant volume and constant flux gravity currents. It was found that deviations of the predicted cloud footprint area change with time from the theoretical were acceptably small, if one models the frictional forces between cloud and ambient air, neglecting the Richardson dependence.The present model has also been validated in widely different experimental conditions such as the Thorney Island instantaneous isothermal releases 8 (unobstructed) and 21 (with semicircular fence), the EEC-55 two-phase propane experiment (with and without linear fence), the Desert Tortoise 4 two-phase ammonia experiment and the Hamburg DAT-638 instantaneous inclined plate experiment and the model predictions were found in reasonable agreement with the experimental data. PMID- 12719148 TI - Production of ultrahigh purity copper using waste copper nitrate solution. AB - The production of ultrahigh purity copper (99.9999%) by electrolysis in the presence of a cementation barrier has been attempted employing a waste nitric copper etching solution as the electrolyte. The amount of copper deposited on the cathode increased almost linearly with electrolysis time and the purity of copper was observed to increase as the electrolyte concentration was increased. At some point, however, as the electrolyte concentration increased, the purity of copper decreased slightly. As the total surface area of cementation barrier increased, the purity of product increased. The electrolyte temperature should be maintained below 35 degrees C in the range of investigated electrolysis conditions to obtain the ultrahigh purity copper. Considering that several industrial waste solutions contain valuable metallic components the result of present study may support a claim that electrowinning is a very desirable process for their treatment and recovery. PMID- 12719149 TI - In situ heavy metal attenuation in landfills under methanogenic conditions. AB - The purpose of this research was to determine the fate and behavior of heavy metals co-disposed with municipal waste under methanogenic conditions. Two landfill simulating reactors, one with leachate recirculation and the other without, were operated in a constant room temperature at 32 degrees C. These reactors were filled with shredded and compacted municipal solid waste having a typical solid waste composition of Istanbul region. After the onset of the methanogenic conditions, the selected heavy metals including iron, copper, nickel, cadmium and zinc were added according to the amounts suggested for co disposal under the directives of the Turkish Hazardous Waste Control Regulations. The results of the experiments indicated that about 90% of all heavy metals were precipitated from the reactors within the first 10 days due to the establishment of highly reducing environment and the formation of sulfide from sulfate reduction which provided heavy metal precipitation. No inhibition to the biological stabilization was observed. PMID- 12719150 TI - Sorption and diffusion of chlorinated aliphatic hydrocarbon penetrants into diol chain extended polyurethane membranes. AB - Sorption and diffusion of a number of chlorinated alkanes through a diol chain extended polyurethane (PU) membranes have been investigated at 25, 40 and 60 degrees C, based on an immersion weight gain method. From the sorption result, the diffusion (D) and permeation (P) coefficients of halogenated hydrocarbon penetrants have been calculated. Molecular transport data depends on membrane solvent interactions, size of the penetrants, temperature and also morphology of the chain extended PUs. The temperature dependence of the transport coefficient has been used to estimate the activation parameters for the process of diffusion (E(D)) and permeation (E(P)) from the Arrhenius plots. Furthermore, the sorption results have been interpreted interms of the thermodynamic parameters such as enthalpy and entropy. PMID- 12719151 TI - Organic solvents permeation through protective nitrile gloves. AB - Effective diffusion coefficients for the permeation of several organic solvents through nitrile gloves were experimentally investigated using the American Society for Testing and Materials (ASTM) F-739 test cell method. Fick's effective diffusion coefficients for benzene, toluene, ethyl benzene, xylene, and styrene were estimated to be 0.61+/-0.02, 0.50+/-0.06, 0.27+/-0.02, 0.31+/-0.03, and 0.21+/-0.03 (x10(-6)cm(2)/s), respectively. These results were comparable to results found by others using different permeation models. Using a transient mass diffusion equation and appropriate initial and boundary conditions, the effective diffusion coefficients were found to adequately simulate the concentration profiles of the organic solvents in the collection chamber during the permeation test. The effective diffusion coefficients were found to be inversely correlated to the molecular weight of the compounds. The results of this study can be used to estimate the exposure to workers using nitrile gloves. PMID- 12719152 TI - A titania thin film annular photocatalytic reactor for the degradation of polycyclic aromatic hydrocarbons in dilute water streams. AB - An external lamp, annular photocatalytic reactor with titania immobilized on a quartz tube was used to degrade two polycyclic aromatic hydrocarbons (PAHs), viz. phenanthrene (PHE) and pyrene (PYR) from a dilute water stream. The thin film geometry was used to obtain both the mass transfer coefficients and intrinsic reaction rate constants for the two compounds on immobilized titania (Degussa P 25) particles. Beyond a feed velocity of 7 cmmin(-1), the conversion was solely reaction rate controlled and was not subjected to mass transfer limitations from the aqueous phase to the immobilized titania film. The overall reaction rate constant was independent of the feed concentration as large as the saturation aqueous solubility of the two compounds. However, the conversion was dependent on the ultraviolet (UV) light illumination intensity at the reactor. The quantum efficiency ranged from 3.7 x 10(-5) to 2.7 x 10(-4) which was somewhat low because of the very low aqueous concentrations of the chemicals. The overall reaction rate constant was 1.6 times larger for pyrene than for phenanthrene. Seven reaction intermediates were identified for the conversion of phenanthrene, while for the degradation of pyrene two intermediates were identified. The presence of the phthalate ester as an intermediate product in the degradation of both PAHs indicates the presence of a quinone in both cases which degrades to the products CO(2) and H(2)O, along with other stable intermediates. Mass balance in a batch reactor showed that only 28.6-40.1% of phenanthrene is mineralized to CO(2) in 1-3h of reaction although 35-67% of the parent compound has disappeared, confirming that a substantial fraction of the parent compound has been converted to stable intermediates that remain in the reactor. A plausible mechanism based on these observations is proposed. PMID- 12719157 TI - Elastic cord-induced ocular injuries. AB - PURPOSE: To evaluate the incidence and severity of ocular injuries caused by elastic cords. PATIENTS AND METHOD: A retrospective review of patients' records from the Emergency Department was conducted to identify cases of elastic cord induced ocular injuries. Over a 67-month period starting in October 1996, 28 patients (24 males and 4 females, mean age 38.6 years) with elastic cord associated ocular trauma were identified. Patient demographics, mechanism of injury, presenting and final visual acuity, ocular injuries, medical and surgical intervention and length of follow-up were noted. RESULTS: The most common anterior and posterior segment injuries were hyphaema/microhyphaema (71.4%) and commotio retinae (60.7%), respectively. The most severe injuries sustained were corneal or scleral lacerations (7.1%), formation of retinal tears (10.7%) or retinal detachment (3.6%), lens subluxation (7.1%) and traumatic cataract (7.1%). Eight patients (28.6%) were admitted for surgical or medical treatment. Mean visual acuity was 20/40 on presentation and 20/25 at the end of the follow-up period. CONCLUSIONS: Elastic cords may cause a wide range of ocular traumas of varying severity. Appropriate design modifications that would prevent the spontaneous release of their hooks are likely to make their use significantly safer. PMID- 12719158 TI - Current spinal board usage in emergency departments across the UK. AB - The spinal board is widely used as a means of extrication and efficient transport during the pre-hospital phase of trauma management. A number of concerns have been raised regarding its subsequent usage once the patient arrives in the emergency department. We undertook a telephone study of 100 A+E departments in the United Kingdom to ascertain current spinal board usage. Our study demonstrated great variability in practice across the UK and a marked lack of on going audit or defined protocols governing spinal board usage following the pre hospital phase of trauma management. PMID- 12719159 TI - Blunt cervical spine injuries in Scotland 1995-2000. AB - BACKGROUND: In the Accident and Emergency Department, the management of patients who have sustained head injuries (HI) is often made more complicated by the suspicion of a cervical spine injury (CSI). This study aimed to evaluate the incidence of CSI in patients sustaining blunt head injuries in a Scottish population. METHODS: Retrospective analysis of prospectively collected data for a 5-year period from the Scottish Trauma Audit Group (STAG) database. Logistic regression and other comparisons were used to investigate the relationship between Glasgow coma score/scale (GCS) and the incidence of CSI. RESULTS: A total of 5154 patients met the criteria for the study and 273 of the HI patients had associated CSI giving an overall incidence of 5.3%. Patients presenting with GCS of 3 were almost three times more likely to have a CSI compared to patients with an initial GCS of 4 or more (12.5% versus 4.4%, chi(2)=62.9, d.f.=1, P<0.001). When patients with GCS of 3 were excluded, there was no evidence of an increase in the incidence of CSI with a lower GCS (logistic regression chi(2)=0.09, d.f.=1, P=0.75). CONCLUSION: The risk of CSI in patients with blunt head trauma and an admission GCS of > or =4 does not decrease as GCS increases. Patients with blunt head injuries who present with a GCS of 3 are much more likely to have a concomitant CSI. The overall incidence of 5.3% compares with published series from other countries. PMID- 12719160 TI - Ultimate tensile strength of a Leeds-Keio/autograft ACL reconstruction utilizing PLLA tibial staple fixation. AB - An ovine model of anterior cruciate ligament reconstruction was utilized to evaluate the biomechanical and histological response of a polylactic acid tibial fixation staple (Gunze Ltd., Japan/Zimmer, Japan). This was performed in a comparison with metallic staples, currently utilized for this procedure. The prosthesis consisted of autograft combined with a Leeds-Keio (L-K) ligament. Early post-operative mobilization was followed by retrieval of specimens at 6, 12 and 24 weeks post-reconstruction. Evaluation of the mechanical characteristics of the graft reconstructions (tensile strength) showed no significant differences (P>0.05) between the staple types for each time period. The histological response to the polylactic acid staple was minimal over the time period studied, with no adverse tissue reactions observed. The mode of reconstruction failure was observed to change with time (P<0.05) presumably as the graft characteristics alter. Overall the absorbable staples performed at a comparable level with the metallic staples within the scope of the study. PMID- 12719161 TI - Meniscal tears sustained awaiting anterior cruciate ligament reconstruction. AB - We reviewed 68 patients who underwent anterior cruciate ligament (ACL) reconstruction after initial EUA & arthroscopy in the knee unit of a United Kingdom district general hospital. Mean time between injury and ACL reconstruction surgery was 23.3 months with the incidence of meniscal tear at reconstruction being 67.6%. In this series 10.3% of patients sustained a meniscal tear in the delay period between arthroscopy and reconstruction surgery. Meniscal tear is strongly associated with ACL rupture and also a poorer outcome following reconstruction surgery. The delay in diagnosis and wait for reconstruction surgery that patients experience are potentially worsening surgical outcomes in the anterior cruciate deficient knee. PMID- 12719162 TI - The Marchetti-Vicenzi nail. A DGH experience. AB - From July 1993 to September 1997, 28 nailings were done on 26 patients using the Marchetti-Vicenzi flexible nail. The minimum length of follow-up was 1 year. All the patients were examined clinically and radiographically. All the 19 tibiae united. Of the nine femur fractures, one required exchange nailing. Complications were delayed union, mal-union, shortening and infection. The complication rate was 3/19 for the tibia and 2/19 for the femur. The mean operative time and the mean fluoroscopy time for the tibia nailings was 36 and 0.22 min and for the femur nailings was 39 and 0.20 min, respectively. This was much lower than that for Russell-Taylor nails. PMID- 12719163 TI - Femoral exchange nailing for aseptic non-union: not the end to all problems. AB - We report the results of a single centre prospective study of exchange nailing for aseptic non-union of a femoral fracture. Eighteen patients with 19 aseptic femoral non-unions had exchange nailing performed in our institution. We collected data on mechanism of injury, original fracture type, and indication for exchange nailing, further surgery and major complications. In 11 non-unions (58%), the exchange nail procedure alone resulted in fracture union with a mean time to radiographic union of 9 months (range 3-24 months). The non-union did not heal in five patients, two patients developed an infected non-union, and one patient required dynamisation of the exchange nail. Fracture healing was eventually achieved in 18 non-unions (95%). Complications following exchange nailing occurred in 11 fractures (58%), in which further surgery was required (four repeat exchange nailings, two Ilizarov frame applications and five nail removals). The role of reamed exchange nailing in the treatment of femoral non union needs to be re-evaluated. Although fracture healing is eventually achieved in most patients, a significant number of them required additional surgery to achieve union or to deal with complications arising from the exchange nailing. PMID- 12719164 TI - Electrical treatment of tibial non-union: a prospective, randomised, double-blind trial. AB - Electrical stimulation in the treatment of bony non-union has been used in different forms for many years. However, there is still a lot of uncertainty about its efficacy. We, therefore, undertook a prospective, randomised, double blind trial to try and determine its effect. Over a period of 5 years, 34 consecutive patients with a tibial non-union met our "criteria for inclusion". Each patient had an oblique fibular osteotomy, followed by a unilateral external fixator. They were then randomly allocated one of two machines. Group 1, the active group, received electrical stimulation from an active machine. Group 2, the dummy group, had an identical machine but without any current passing through the active coils. They were then followed up for 6 months and evaluated clinically and radiologically for bony union. Unfortunately, there was by chance, an imbalance in smoking habit between the two groups. The union rate in the subgroup that smoked was 75% (6/8) in the active group as compared to 46% (6/13) in the dummy group. The active group of non-smokers had 100% (10/10) union rate, compared to 67% (2/3) in the dummy group. Overall 24 out of the 34 patients progressed to union. Out of 18, 16 (89%) in the active group showed bony union as compared to 8/16 (50%) in the dummy. There was, thus, a statistically significant positive association between tibial union and electrical stimulation (odds ratio 8, 95% CI: 1.5-41, P=0.02). PMID- 12719165 TI - Experimental model for a new distal locking aiming device for solid intramedullary tibia nails. AB - A distal locking device for the solid intramedullary tibial nails has been tried on bone models. This device allows simple and accurate insertion of distal locking screws with minimum radiation exposure to the patient and the surgeons. It also reduces the operation time. The aim of this experiment was to test the accuracy of the new aiming device and to measure the radiation exposure time during the procedure. PMID- 12719166 TI - Lightning and Lichtenberg figures. PMID- 12719167 TI - Immobilisation of spinal fractures in patients with ankylosing spondylitis. Two case reports. PMID- 12719168 TI - Case report: metaphyseal osteolysis around a titanium reconstruction nail. PMID- 12719169 TI - Radiological appearance as a meniscal ossicle develops: a case report and review of literature. PMID- 12719170 TI - Intra-articular migration of anterior cruciate ligament graft fixation presenting as a locked knee. PMID- 12719171 TI - Spontaneous healing of a 14 cm diaphyseal cortical defect of the tibia. PMID- 12719172 TI - Maintaining reduction during unreamed nailing of a segmental tibial fracture: the use of a Farabeuf clamp. PMID- 12719173 TI - The operative management of displaced intra-articular fractures of the calcaneum, a two-centre study using defined protocols. PMID- 12719175 TI - Case report--fracture of the occipital condyle. PMID- 12719177 TI - Vulnerable places: contextualizing health practices. PMID- 12719178 TI - From space to place in contemporary health care. PMID- 12719179 TI - Hidden places, uncommon persons. AB - Specialized hospital units recently created to house and maintain ventilator or other technology-dependent persons in the United States are new cultural forms that enable beings who are neither fully alive, biologically dead, nor "naturally" self-regulating, yet who are sustained by modern medical practices, to exist. These institutions both fabricate and complicate the persons who are patients there through surveillance and maintenance of their conditions. This article concerns the relationship of person to place when the consciousness of an individual, considered to be the essence of personhood in the modern Western philosophical tradition, is problematic because the person resides in a technologically produced border zone between life and death. The article explores the ways in which place and person become implicated one another: first, how consciousness and thus personhood is assessed and negotiated through the inter subjective knowledge of hospital staff; second, how that knowledge is tied to the particular situate-dness of patients; and third, how embodiment itself-the reflexive knowledge of the-self-in-the-body-is perceived as emplaced in social and spatial relations. PMID- 12719180 TI - Space, place and movement as aspects of health care in three women's prisons. AB - This paper focuses on prison as a place in which the prisoner seeks health and health services. Drawing on the work of Henri LeFebvre, Edward Casey, Jeffrey Malpas, and Michel Foucault, a spatial analysis examines the constitutive roles of movement, social structure, and power in determining the prisoner's access to health care. The research methodology utilizes quantitative and qualitative analysis of women prisoners' attempts to get treatment for their health problems. The narratives of these often-failed attempts construct prison as a place where health care access is continually thwarted by rules, custodial priorities, poor health care management, incompetence, and indifference. Analysis of spatial practices, representations of space, and spaces of representation demonstrate the imposition of structural ordering, its naturalization, and the role of narrative in questioning the order, thereby creating possibilities for imaginary and real places where the prisoners' health needs can be met. Simultaneously, this analysis illuminates basic ethical questions about the limitations of human connection and medical caring in prison settings, regardless of the personal motivation of the caregiver. PMID- 12719181 TI - Bioethics and rural health: theorizing place, space, and subjects. AB - The field of bioethics has been criticized for its universalizing tendencies, attributed in a large part to its foundations in moral philosophy and the level of abstraction of much bioethical discourse. Efforts to particularize bioethics have included the "turn toward casuistry", the emergence of feminist and disability rights critiques of mainstream bioethics, and ethnographic contributions that examine the situatedness of ethical acts, practices, and meanings in local contexts. Such work introduces into bioethics dimensions of space, place, and time; nonetheless, these remain relatively unexplored as constitutive elements and/or influences of the phenomena of ethics discourse and ethics-related practices. Drawing from an ethnographic study of genetics in rural health, this paper presents a sociological discussion of space/time and bioethics through examination of rural health settings. Issues raised include intersections of spatial and power relations, socio-spatial gradients of expertise, and socio spatial dimensions of ethics knowledge and practices within medical settings. PMID- 12719182 TI - From home to hospital and back again: economic restructuring, end of life, and the gendered problems of place-switching health services. AB - Economic restructuring in the health services industry in the USA exemplifies general patterns of economic change propelled by neoliberalism, especially industry privatization, diminished social services, and dependence on "flexible" labor and management regimes. Combined with the widespread entry of women into the labor force, an aging population, and minimal assistance for high quality long-term care at the end of life, these economic and social conditions raise a set of difficult policy questions for health services planning. Set in these broad contexts, this paper situates access to and experience of health services in the home, the hospital, and nursing facility, to demonstrate how economic changes have relocated and redefined health services in ways that distinctively impact how people experience the places where they receive care. This place switching of health services externalizes costs of subacute and "daily life care" (the so-called custodial care) to the sphere of the individual, their family, and communities. The theoretical analysis uses current geographical and philosophical approaches to place and space, and considers the tensions between institutionally managed health care space, and the patient's experience of receiving health services in place. The place/space dilemma of health services provision is examined through several interrelated subjects: long-term care at the end of life, gendered characteristics of care giving, the limitations of Medicare and Medicaid, historical changes in hospital length of stay, the restructuring of nursing practices, and the "no-care zone". The analysis is based on examples of stroke and incontinence care to demonstrate the importance of considering place and space issues in health care planning. PMID- 12719183 TI - Placing private health care: reading Ascot Hospital in the landscape of contemporary Auckland. AB - The closing years of the 20th century were a time in New Zealand dominated by health care reforms inspired by neo-liberal ideology. The result has been changing geographies of public and private health care providers and the evolution of a new discourse of health care. Ascot Integrated Hospital, situated in the affluent Auckland suburb of Remuera, opened in 1999, reflecting and projecting this new discourse. It is a pioneer, competing for patient patronage in a contracting market for surgical and medical providers. In this paper we survey the recent history of private hospital developments in New Zealand, then more closely consider the Ascot, a hospital that has deployed language to construct itself and its achievements in the public imagination. Given the context of an extremely competitive environment for private patients, this construction glamorises medicine and links healing with a contrived place. We conclude that texts associated with the Ascot provide a useful vehicle for advancing cultural geographies of health care and ideas of the place of hospitals in western capitalist countries. PMID- 12719184 TI - Distal nursing. AB - This paper considers the spatial dynamics of nurse-patient relationships within hospitals, primarily in the USA, under conditions of organizational restructuring, and situates them within social theoretical perspectives on space. As a human practice to which relationship is considered essential, nursing depends upon sustaining an often taken-for-granted proximity to patients. But hospital nursing, I argue in this paper, is increasingly constrained by spatial structural practices that disrupt relationship and reduce or eliminate such proximity. Three kinds of proximity are threatened: physical, narrative, and moral. Examining these proximities through a place-space lens suggests that nursing is increasingly "distal" to patient care. There are potentially dangerous implications in this loss of proximity. PMID- 12719185 TI - Locating gene-environment interaction: at the intersections of genetics and public health. AB - Over the past two decades, the applications of genetic and genomic technologies have begun to transform research questions and practices within epidemiology and toxicology, the "core sciences" of public health (Annu. Rev. Public Health 21 (2000) 1). These technologies provide new models and techniques for studying genetic traits, environmental exposures, and gene-environment interaction in the production of human health and illness. This paper explores the consequences of emergent genetic and genomic approaches, their ongoing redefinitions of both genetic and environmental "risks", and their potential implications for public health practice. The central argument of the paper is that the increasing focus on gene-environment interaction directs scientific, biomedical, and public health attention both inward, to the gene/genome, and outward, to particular places. In so doing, studies of gene-environment interaction create a challenging and productive tension-at the same time that bodies are being geneticized (Am. J. Law Med. 17 (1992) 15), they also are emphatically emplaced, located where social and cultural practices come to matter. This tension, this simultaneous movement outward and inward, towards the gene and towards the environment, into the body and into place, opens up a vista into the processes through which culture and biology form a locally and historically situated dialectic (Encounters With Aging: Mythologies of Menopause in Japan and North America, University of California Press, Berkeley, CA, 1993) and raises important questions about the production of health and illness. PMID- 12719186 TI - Bio-medical Topoi--the dominance of space, the recalcitrance of place, and the making of persons. PMID- 12719187 TI - Adaptive Dynamics of the Leg Movement Patterns of Human Infants: I. The Effects of Posture on Spontaneous Kicking. AB - This is the first of two articles in which we describe how infants adapt their spontaneous leg movements to changes in posture or to elicitation of behaviors by a mechanical treadmill. In this article, we compare the kinematics of kicks produced by 3-month-old infants in three postures, supine, angled (45 degrees ), and vertical, and examine the changes in muscular and nonmuscular force contributions to limb trajectory. By manipulating posture we were able to assess the sensitivity of the nascent motor system to changes in the gravitational context. The postural manipulation elicited a distinct behavioral and dynamic effect. In the more upright postures, gravitational resistance to motion at the hip was 4 to 10 times greater than resistance met in the supine posture, necessitating larger muscle torques to drive hip flexion. Kicks produced in the vertical posture showed a reduction in hip joint range of motion and an increase in synchronous joint flexion and extension at the hip and knee. At the same time, hip and knee muscle torques were also more highly correlated in kicks performed in the vertical than in the supine or angled posture. This increased correlation between muscle torques at the hip and knee implicates anatomical and energetic constraints-the intrinsic limb dynamics-in creating coordinated limb behavior out of nonspecific muscle activations. PMID- 12719188 TI - Adaptive Dynamics of the Leg Movement Patterns of Human Infants: II. Treadmill stepping in Infants and Adults. AB - Infant treadmill steps have many temporal and kinematic similarities to adult walking. Kinematic similarities can result from different patterns of underlying torque, however. In this study, we used inverse dynamics to compare the patterns and contributions of active (muscle) and passive (gravity and motion-dependent) torques in the swing phase of treadmill stepping in 7-month-old infants and adults. Results indicated that adults consistently used muscle torque to initiate and terminate swing, but that passive torques accounted for leg motion during most of the swing phase. Infants, in contrast, displayed multiple patterns of torque contributions during swing. In the most frequently occurring infant pattern, muscle torque remained flexor throughout swing and joint reversals were due to the dominant passive gravitational torque. The kinetic data suggest that the temporally and kinematically similar treadmill steps produced by adults and infants do not emanate from a unique set of neural commands to the muscles, but from a flexible interplay between multiple internal as well as external elements. These data suggest that the intrinsic dynamics of the human system provide a medium out of which, given a supportive context, stable patterns can emerge spontaneously. During development, voluntary controlled movement patterns must build on these intrinsic dynamics. PMID- 12719189 TI - Effects of Object Texture on Precontact Movement Time in Human Prehension. AB - Using kinematic data in a precision-grip reaching task, Weir, MacKenzie, Marteniuk, and Cargoe (1991) concluded that prior to contact with an object, its texture does not affect the course of grasping. The present study used their task of reaching for and lifting a slippery-, normal- (polished metal), or rough surfaced dowel. This occurred under the original, blocked condition, in which textures were held constant within a series of trials, and under a new, randomized condition, in which textures varied randomly from trial to trial. Performance was also examined over more extended periods of practice. Reaction time and precontact movement time were directly measured. In contrast to the results of Weir et al., 1991, reaching for the slippery dowel resulted in slower movement time. This effect was found both early and late in practice for the randomized condition; it was found only in late practice for the blocked condition. These effects can be attributed to the greater geometric and dynamic precision required for lifting a slippery object. PMID- 12719190 TI - The Influence of Skill and Intermittent Vision on Dynamic Balance. AB - Two experiments are reported in which expert and novice gymnasts were required to walk across a balance beam as quickly as possible in various vision conditions. In Experiment 1, experts walked faster than novices in all vision conditions, showing the greatest superiority when vision was completely eliminated. Novices were more dependent on vision and were able to maintain their performance as long as a visual sample was available every 250 ms (i.e: 4-Hz samples).The results of Experiment 2 indicate that differences between expert and novice performers in the no-vision condition were not related to the use of a short-term visual representation of the movement environment. Our movement time findings are problematic for specificity of learning models of skill acquisition. As well, film data collected in Experiment 2 were not consistent with models that propose a transition from closed-loop to open-loop control. PMID- 12719191 TI - Motor Performance of Stutterers and Nonstutterers on Timing and Force Control Tasks. AB - Recently it has been suggested that speech and manual timing tasks share a common central process (Franz, Zelaznik, & Smith, 1992): Because stuttering is thought to be related to deficits in motoric processes such as timing, stutterers (n = 15) were compared with a set of age-, education-, and sex-matched nonstutterers on timing and isometric force-production tasks. In the timing tasks, subjects flexed and extended the right index finger at the metacarpophalangeal joint at cycle durations of 600, 500, 400, 300, and 200 ms. In the force-production tasks, subjects generated isometric forces to match target force levels displayed on a cathode-ray tube (CRT) screen. There were five levels of force, ranging from.11 to 7.85 newtons. Overall, there were no differences in timing and force production performance between stutterers and nonstutterers. These results are similar to those obtained recently by Hulstijn, Summers, van Lieshout, and Peters (1992). We suggest that stuttering is not characterized by a general deficit in rhythmic timing. Instead, the motor deficit associated with stuttering should be viewed as speech specific. PMID- 12719192 TI - Feedback-Induced Variability and the Learning of Generalized Motor Programs. AB - As compared with providing extrinsic feedback on each of a set of practice trials, reducing the feedback frequency in various ways facilitates long-term retention. One explanation is that frequent feedback operates proactively on the subsequent trial, inducing excessive variability that degrades learning. We tested this view by giving or not giving feedback "reminders," where the postresponse feedback was given again just before the next attempt at a task. The reminder manipulation was examined in both blocked and randomized practice sequences during the learning of three limb-patterning tasks. Reminder feedback increased response variability during practice in both random and blocked practice. As measured in retention tests, though, feedback reminders degraded learning in random practice, but not in blocked practice. This implies that the frequently found learning advantages of random, as compared with blocked, practice might be due to feedback's facilitation of retrieval operations in blocked practice (as well as in random practice with feedback reminders); such overfacilitation of retrieval operations has been shown to degrade learning. Additional analyses revealed that random, as compared with blocked, practice enhanced the learning of the fundamental pattern of action (generalized motor program) but had little effect on the ability to scale the pattern in amplitude or time (parameterization). PMID- 12719193 TI - Reducing Knowledge of Results About Relative Versus Absolute Timing: Differential Effects on Learning. AB - The purpose of the present experiment was to examine further earlier suggestions that a reduced relative frequency of knowledge of results (KR) can enhance the learning of generalized motor programs (GMPs) but at the same time degrade parameter learning, compared with giving KR after every trial (Wulf & Schmidt, 1989; Wulf, Schmidt, & Deubel, 1993). In contrast to these earlier studies, here KR was given separately for relative timing and absolute timing. Subjects practiced three movement patterns that required the same relative timing but different absolute movement times. KR was provided on 100% or 50% of the practice trials for relative timing or absolute timing, respectively. In retention and transfer tests, the groups that had had 50% KR about relative timing demonstrated more effective learning of the relative-timing structure, that is, GMP learning, than the groups that had had 100% KR about relative timing. The KR frequency had no effect on parameterization during retention; yet, when transfer to a task with a novel overall duration was required, the groups given 100% KR about absolute timing were more accurate in parameterization than the groups provided with 50% KR about absolute timing. Thus, the reduced relative KR frequency enhanced GMP learning but had no beneficial effect, or even a degrading effect, on parameter learning. The differential effects of a reduced KR frequency on the learning of relative timing and absolute timing also provide additional support for the dissociation of GMP and parameterization processes. PMID- 12719194 TI - Meeting the challenge of peripheral arterial disease. PMID- 12719195 TI - The perioperative management of warfarin therapy. PMID- 12719196 TI - Critical issues in peripheral arterial disease detection and management: a call to action. PMID- 12719197 TI - Linking laboratory and pharmacy: opportunities for reducing errors and improving care. AB - A myriad of errors and lost improvement opportunities result from failure of clinical laboratory and pharmacy information systems to effectively communicate. Pharmacotherapy could benefit from enhanced laboratory-pharmacy linkage with respect to (1) drug choice (laboratory-based indications and contraindications), (2) drug dosing (renal or hepatic, blood level-guided adjustments), (3) laboratory monitoring (laboratory signals of toxicity, baseline and ongoing monitoring), (4) laboratory result interpretation (drug interfering with test), and (5) broader quality improvement (surveillance for unrecognized toxicity, monitoring clinician response delays). Linkages can be retrospective or real time. Many organizations could benefit now by linking existing pharmacy and laboratory data. Greater improvement is possible through implementation of electronic order entry with real-time decision support incorporating linked laboratory and pharmacy data. While many guidelines, admonitions, and rules exist regarding drugs and the laboratory, substantial new knowledge and evidence in this area are needed. Focusing on these unmet needs and accompanying logistical challenges is a priority. PMID- 12719198 TI - Perioperative management of patients receiving oral anticoagulants: a systematic review. AB - BACKGROUND: The safety and efficacy of various management strategies for patients receiving oral anticoagulants (OACs) who need to undergo surgery or invasive procedures are unknown. METHODS: We performed a systematic review and synthesis of the English-language literature examining the perioperative management and outcomes of patients receiving long-term OAC therapy. RESULTS: Thirty-one reports were identified. The quality of the identified reports was generally poor; no randomized controlled trials have been performed and duration of follow-up was typically not stated. Overall, 29 thromboembolic events occurred amont 1868 patients (1.6%; 95% confidence interval, 1.0%-2.1%), including 7 strokes (0.4%; 95% confidence interval, 0%-0.7%). Thromboembolic event rates by management strategy were 0.4% (1 of 237) for continuation of OAC, 0.6% (6 of 996) for discontinuation of OAC therapy without administration of intravenous heparin, 0% (0 of 166) for discontinuation of OAC therapy with administration of intravenous heparin, 0.6% (1 of 180) for discontinuation of OAC therapy with administration of low-molecular-weight heparin, and 8.0% (21 of 263) for unspecified or unclear strategies. Major bleeding while receiving therapeutic OAC was rare for dental procedures (0.2% [4 of 2014]), arthrocentesis (0% [0 of 32]), cataract surgery (0% [0 of 203]), and upper endoscopy or colonoscopy with or without biopsy (0% [0 of 111]). CONCLUSIONS: Most patients can undergo dental procedures, arthrocentesis, cataract surgery, and diagnostic endoscopy without alteration of their regimen. For other invasive and surgical procedures, oral anticoagulation needs to be withheld, and the decision whether to pursue an aggressive strategy of perioperative administration of intravenous heparin or subcutaneous low molecular-weight heparin should be individualized. The current literature is substantially limited in its ability to help choose an optimal strategy. Further and more rigorous studies are needed to better inform this decision. PMID- 12719199 TI - Patient attitudes toward continuity of care. AB - BACKGROUND: Concern has been raised about managed care's effects on continuity of patient care, but little is known about how much value patients place on continuity. METHODS: We surveyed 2500 adult patients of a large New England health maintenance organization about their attitudes toward continuity and their willingness to spend additional time or money to maintain continuity with their primary care physician (PCP). RESULTS: Among the 1171 (46.8%) of patients responding, 460 (39.6%) of 1162 patients had had more than one PCP in the previous 5 years. Nearly all patients (1068 [91.5%] of 1167) rated continuity as very important or important; only 26 (2.2%) rated continuity as unimportant or very unimportant. However, only 256 (22.2%) of 1152 patients were willing to drive more than 60 minutes to maintain continuity with their PCP, and only 200 (18.2%) of 1096 would be willing to spend an additional $20 to $40 per month to maintain it. In multivariable analyses, patients were more willing to drive if they were nonwhite (odds ratio [OR], 2.3; 95% confidence interval [CI], 1.4-3.6), older than 50 years (OR, 1.7; 95% CI, 1.2-2.4), or had less than a college education (OR, 1.6; 95% CI, 1.2-2.2). Patients who had been forced to change PCPs when their physician moved away were less willing to drive (OR, 0.5; 95% CI, 0.3 0.8) or spend more money (OR, 0.7; 95% CI, 0.5-1.0) to maintain continuity. CONCLUSIONS: Most patients in this sample indicated that continuity of care was important to them, but reported being unwilling to spend much additional personal time or money to maintain continuity with their current PCP. Nevertheless, an important subset of older and more vulnerable patients reported being more willing to pay to maintain continuity. PMID- 12719200 TI - The outpatient bleeding risk index: validation of a tool for predicting bleeding rates in patients treated for deep venous thrombosis and pulmonary embolism. AB - BACKGROUND: Long-term anticoagulation prevents recurrent thrombosis in patients with idiopathic deep venous thrombosis or pulmonary embolism, but with a risk of clinically important so-called major bleeding. Physician- and patient-based decisions on the optimal duration of therapy are sensitive to the bleeding risk. The Outpatient Bleeding Risk Index potentially provides a means of calculating the potential risk of bleeding using easily elicited clinical findings, but, to our knowledge, the authors of the index have provided the only published validation of it. We sought to determine the accuracy of the index in our population of patients. METHODS: We prospectively applied the Outpatient Bleeding Risk Index to consecutive patients in our clinic who had been objectively diagnosed as having pulmonary embolism or deep venous thrombosis and who were about to undergo standard therapy. Standard therapy consisted of a minimum of 5 days of low-molecular-weight heparin therapy overlapped with warfarin sodium therapy, and continuation of warfarin therapy for at least 3 months, with a target international normalized ratio of 2.5. Patients were placed in 3 risk groups (low, moderate, or high), as defined by the index. The survival curves of the groups, using major hemorrhages as the events, were then compared by the log rank test. RESULTS: Bleeding rates were lower than expected, but the index did discriminate between low- and moderate-risk groups (P =.03, log-rank test). The rate of major hemorrhage per 100 person-years was 0% (95% confidence interval, 0% 2.8%) in the low-risk group and 4.3% (95% confidence interval, 1.1%-11.1%) in the moderate-risk group. The rate in the high-risk group could not be defined because only 2 patients were at high risk. CONCLUSION: The Outpatient Bleeding Risk Index discriminates between low- and moderate-risk patients, and could be used to guide decisions on the optimal duration of anticoagulant therapy. PMID- 12719201 TI - Disparities in use of lipid-lowering medications among people with type 2 diabetes mellitus. AB - BACKGROUND: People with diabetes are at high risk for cardiovascular events regardless of known heart disease. Physicians may underrecognize the excess cardiovascular risk conferred by diabetes alone, without a recent cardiovascular event. Other disparities in the receipt of lipid-lowering medications (LLMs) may exist. METHODS: We studied veterans with diabetes in fiscal years 1998 and 1999 cross-sectionally. We used administrative data (demographic information, International Classification of Diseases, Ninth Revision [ICD-9] codes, utilization information, medications, and laboratory tests) to evaluate associations between use of LLMs and age, ethnicity, sex, marital status, Charlson Index, heart disease ICD-9 codes, oral agents and insulin, hospitalization status, and low-density lipoprotein cholesterol levels. We constructed separate logistic regression models to evaluate associations between low-density lipoprotein cholesterol and similar predictor variables. RESULTS: Odds ratios were similar in both years. For fiscal year 1999, patients without recent ICD-9 codes in their administrative data indicating heart disease were 0.35 times less likely to be given LLMs than those with such codes. Individuals older than 75 years were 0.65 times less likely to be given LLMs than those younger than 65 years. African Americans were 0.72 times less likely than whites to be given LLMs. In fiscal years 1999 and 1998, 27% and 36% of individuals given LLMs had low-density lipoprotein cholesterol levels higher than 130 mg/dL (3.37 mmol/L). CONCLUSIONS: Veterans with diabetes but no recently coded heart disease, older individuals, and African Americans could benefit from programs targeted to introduce LLMs. Up to one third of individuals given LLMs remained above the target level of 130 mg/dL for low-density lipoprotein cholesterol. PMID- 12719202 TI - The economic consequences of irritable bowel syndrome: a US employer perspective. AB - BACKGROUND: The objective of this study was to measure the direct costs of treating irritable bowel syndrome (IBS) and the indirect costs in the workplace. This was accomplished through retrospective analysis of administrative claims data from a national Fortune 100 manufacturer, which includes all medical, pharmaceutical, and disability claims for the company's employees, spouses/dependents, and retirees. METHODS: Patients with IBS were identified as individuals, aged 18 to 64 years, who received a primary code for IBS or a secondary code for IBS and a primary code for constipation or abdominal pain between January 1, 1996, and December 31, 1998. Of these patients with IBS, 93.7% were matched based on age, sex, employment status, and ZIP code to a control population of beneficiaries. Direct and indirect costs for patients with IBS were compared with those of matched controls. RESULTS: The average total cost (direct plus indirect) per patient with IBS was 4527 dollars in 1998 compared with 3276 dollars for a control beneficiary (P<.001). The average physician visit costs were 524 dollars and 345 dollars for patients with IBS and controls, respectively (P<.001). The average outpatient care costs to the employer were 1258 dollars and 742 dollars for patients with IBS and controls, respectively (P<.001). Medically related work absenteeism cost the employer 901 dollars on average per employee treated for IBS compared with 528 dollars on average per employee without IBS (P<.001). CONCLUSION: Irritable bowel syndrome is a significant financial burden on the employer that arises from an increase in direct and indirect costs compared with the control group. PMID- 12719203 TI - A clinical prediction rule to identify patients with atrial fibrillation and a low risk for stroke while taking aspirin. AB - BACKGROUND: We sought to derive and internally validate a simple and easily applied clinical prediction rule to identify patients with nonvalvular atrial fibrillation (AF) whose stroke risk while taking aspirin is, irrespective of age, low enough that oral anticoagulation therapy is unnecessary. METHODS: We included 2501 patients with AF treated with aspirin during participation in 6 clinical trials. Patients were randomly divided into derivation and validation sets. Recursive partitioning was used to identify patients in the derivation set whose risk for stroke (ischemic or hemorrhagic) or transient ischemic attack was comparable to that observed in an age- and sex-matched cohort from the Framingham Heart Study. The derived prediction rules were tested on the validation set. RESULTS: Overall, 166 patients (6.6%) had an event during 4688.6 person-years (PYs) of observation for an incident rate of 3.5 events per 100 PYs. Patients in the derivation set classified as low risk (no previous stroke or transient ischemic attack, no treated hypertension or systolic blood pressure equal to or exceeding 140 mm Hg, no symptomatic coronary artery disease, and no diabetes) experienced 1.0 events per 100 PYs, compared with an age- and sex-matched rate of 1.2 events per 100 PYs. In the validation set, low-risk patients experienced 1.1 events per 100 PYs (expected rate of 1.2 events per 100 PYs). Low-risk patients made up 24% of the cohort and 16% of patients older than 75 years. Low-risk patients who were randomized to therapeutic oral anticoagulation therapy experienced 1.5 events per 100 PYs. CONCLUSION: Irrespective of age, patients with AF and none of these 4 clinical features and who take aspirin have stroke rates comparable to those of age-matched community cohorts and would not benefit substantially from anticoagulation. PMID- 12719204 TI - Time and money: a retrospective evaluation of the inputs, outputs, efficiency, and incomes of physicians. AB - BACKGROUND: Physicians' concerns with the health care system focus on having less time with their patients and needing to work harder to maintain incomes. We sought to determine whether physicians are working longer hours and whether their incomes are declining. METHODS: Using survey data, we conducted a retrospective analysis of physician inputs, outputs, efficiency, and incomes for generalists, general internists, general surgeons, pediatricians, and obstetrician gynecologists from 1987 to 1998. RESULTS: Physician inputs (as measured by the average hours worked in professional activities) showed little absolute change across specialties over time. Outputs (as measured by the total number of patient visits per week) decreased between 9% and 28%, depending on the specialty. Efficiency (the proportion of time spent in direct patient care and the amount of time spent during a typical office visit) remained stable over the time examined. Consumer price index inflation-adjusted annual incomes increased considerably over the time period examined (42% for general internists, 28% for pediatricians, 13% for generalists, and 8% for general surgeons); only obstetricians gynecologists showed a net loss of annual income when adjusting for inflation (a 6% loss). CONCLUSIONS: Our findings do not confirm the prevailing concern that physicians are working harder or longer or that their incomes are declining, but they offer an explanation of how physicians are maintaining incomes without increasing work inputs. There is a great deal of dissatisfaction with the health care system among physicians; exploration of perceptual reasons for that dissatisfaction may outline a course of action needed to resolve it. PMID- 12719205 TI - Central nervous system active medications and risk for fractures in older women. AB - BACKGROUND: Use of central nervous system (CNS) active medications may increase the risk for fractures. Prior studies are limited by incomplete control of confounders. METHODS: To determine whether use of CNS active medications, including benzodiazepines, antidepressants, anticonvulsants, and narcotics, increases fracture risk in elderly, community-dwelling women, we examined use of these 4 categories of medications in a cohort of 8127 older women and followed the participants prospectively for incident nonspine fractures, including hip fractures. Current use of CNS active medications was assessed by interview with verification of use from containers between 1992 and 1994 and between 1995 and 1996. Use was coded as a time-dependent variable. Incident nonspine fractures occurring after the initial medication assessment until May 31, 1999, were confirmed by radiographic reports. RESULTS: During an average follow-up of 4.8 years, 1256 women (15%) experienced at least one nonspine fracture, including 288 (4%) with first hip fractures. Compared with nonusers, women taking narcotics (multivariate hazard ratio [HR], 1.40; 95% confidence interval [CI], 1.06-1.83) and those taking antidepressants (multivariate HR, 1.25; 95% CI, 0.99-1.58) had increases in the risks for any nonspine fractures. Women taking tricyclic antidepressants and those using selective serotonin reuptake inhibitors (SSRIs) had similar fracture rates. There were no independent associations between benzodiazepine use or anticonvulsant use and risk for nonspine fracture. Women taking antidepressants compared with nonusers had a 1.7-fold increase in the risk for hip fracture (multivariate HR, 1.65; 95% CI, 1.05-2.57). We did not observe independent associations between use of any of the other 3 classes of CNS active medications and risk of hip fracture. CONCLUSIONS: Community-dwelling older women taking narcotics have an increased risk for any nonspine fracture, and those taking antidepressants have a greater risk for nonspine fractures, including hip fracture. Rates of fracture were similar in women taking tricyclic antidepressants and those using SSRIs. Benzodiazepine use and anticonvulsant use were not independently associated with an increased risk of nonspine fractures, including hip fracture. PMID- 12719206 TI - The role of adherence on the effectiveness of nonpharmacologic interventions: evidence from the delirium prevention trial. AB - BACKGROUND: The impact of adherence on outcome for a nonpharmacologic intervention strategy has not been previously examined. OBJECTIVE: To examine the impact of level of adherence on effectiveness of the intervention strategy in a large clinical trial of nonpharmacologic interventions to prevent delirium. METHODS: The subjects included 422 consecutive patients 70 years or older admitted to the medicine service at a university hospital. The intervention protocols were targeted toward 6 delirium risk factors. The primary outcome was new-onset delirium during hospitalization. RESULTS: During 9882 patient-days, complete adherence rates for individual intervention protocols ranged from 10% for the sleep protocol to 86% for the orientation protocol. The rate of complete adherence with all protocols was 57%, and combined partial and complete adherence was 87%. Higher levels of adherence resulted in lower delirium rates, with a significant graded effect, for orientation, mobility, and therapeutic activities protocols, and for the composite adherence measure. After controlling for potential confounding variables, such as illness severity, comorbidity, baseline delirium risk, and functional status, adherence continued to demonstrate a consistently strong and significant protective effect against delirium (adjusted odds ratio, 0.69; 95% confidence interval, 0.56-0.87). Patients in the highest adherence group demonstrated an 89% reduction in delirium risk compared with patients in the lowest group. CONCLUSIONS: Adherence played an important independent role in the effectiveness of a nonpharmacologic multicomponent intervention strategy. Higher levels of adherence resulted in reduced rates of delirium in a directly graded fashion, with extremely low levels of delirium in the highest adherence group. Thus, adherence must be ensured in nonpharmacologic interventions to optimize effectiveness. PMID- 12719207 TI - Fibrinolytic therapy in patients 75 years and older with ST-segment-elevation myocardial infarction: one-year follow-up of a large prospective cohort. AB - BACKGROUND: Fibrinolytic therapy reportedly may not be beneficial in acute ST segment-elevation myocardial infarction (STEMI) in patients who are 75 years and older. METHODS: The association between fibrinolytic therapy and 1-year mortality and bleeding complications in an unselected large cohort of patients with STEMI was evaluated by means of propensity and Cox regression analysis adjusting for multiple factors known to influence fibrinolytic therapy as well as survival. The Register of Information and Knowledge About Swedish Heart Intensive Care Admissions recorded every patient admitted to a coronary care unit in 64 hospitals during 1995 through 1999. One-year mortality was obtained by merging with the National Cause of Death Register. RESULTS: A total of 6891 patients 75 years and older with first registry-recorded STEMI were included, of whom 3897 received fibrinolytic therapy and 2994 received no such treatment. Fibrinolytic therapy was associated with a 13% adjusted relative reduction in the composite of mortality and cerebral bleeding complications after 1 year (95% confidence interval, 0.80-0.94; P =.001). This effect seemed homogeneous among all subgroups based on age, sex, coronary risk factors, and previous disease manifestations. CONCLUSIONS: Fibrinolytic therapy in patients with STEMI who are 75 years and older is associated with a reduction in the composite of mortality and cerebral bleedings after 1 year. These results from an unselected coronary care unit population support the use of fibrinolytic therapy in elderly patients. PMID- 12719208 TI - Unnecessary use of antimicrobials in hospitalized patients: current patterns of misuse with an emphasis on the antianaerobic spectrum of activity. AB - BACKGROUND: Unnecessary use of antimicrobials contributes to the emergence and dissemination of antimicrobial-resistant nosocomial pathogens in part through elimination of normal anaerobic bacterial flora that inhibit overgrowth of pathogenic microorganisms. METHODS: A prospective observational study was conducted in a 650-bed, university-affiliated hospital. All adult nonintensive care inpatients for whom new antimicrobials were prescribed during a 2-week period were monitored throughout their hospitalization. We examined how often antimicrobials, in particular those with antianaerobic activity, were used unnecessarily. The reasons for unnecessary therapy were assessed and common patterns of unnecessary use were identified. RESULTS: A total of 1941 antimicrobial days of therapy were prescribed for 129 patients. A total of 576 (30%) of the 1941 days of therapy were deemed unnecessary. The most common reasons for unnecessary therapy included administration of antimicrobials for longer than recommended durations (192 days of therapy), administration of antimicrobials for noninfectious or nonbacterial syndromes (187 days of therapy), and treatment of colonizing or contaminating microorganisms (94 days of therapy). Antianaerobic agents accounted for 203 (35%) of the 576 unnecessary antimicrobial days of therapy, and these agents were also frequently prescribed (98 days of therapy) when equally efficacious alternative regimens with minimal antianaerobic activity were available. CONCLUSIONS: In our institution, hospitalized patients frequently received unnecessary antimicrobial therapy, and antianaerobic agents were often prescribed when this spectrum of activity was not indicated. PMID- 12719209 TI - Associations between body mass index and risks in elderly patients with a first ever acute ischemic stroke. PMID- 12719210 TI - Low-molecular-weight heparin for anticoagulation during continuous venovenous hemofiltration. PMID- 12719211 TI - Medication errors detected in infusions. PMID- 12719212 TI - Does metformin interfere with thiamine? PMID- 12719213 TI - Subtle shades of permeability. PMID- 12719214 TI - The pH-dependent stability of wild-type and mutant transthyretin oligomers. AB - A reduction in pH is known to induce the disassociation of the tetrameric form of transthyretin and favor the formation of amyloid fibers. Using continuum electrostatic techniques, we calculate the titration curves and the stability of dimer and tetramer formation of transthyretin as a function of pH. We find that the tetramer and the dimer become less stable than the monomer as the pH is lowered. The free energy difference is 13.8 kcal/mol for dimer formation and 27 kcal/mol for tetramer formation, from the monomers, when the pH is lowered from 7 to 3.9. Similar behavior is observed for both the wild-type and the mutant protein. Certain residues (namely Glu-72, His-88, His-90, Glu-92, and Tyr-116), play an important role in the binding process, as seen by the considerable pK(1/2) change of these residues upon dimer formation. PMID- 12719215 TI - The effect of protein conformational flexibility on the electronic properties of a chromophore. AB - In this paper we address the question of how a protein environment can modulate the absorption spectrum of a chromophore during a molecular dynamics simulation. The effect of the protein is modeled as an external field acting on the unperturbed eigenstates of the chromophore. Using a first-principles method recently developed in our group, we calculated the perturbed electronic energies for each frame and the corresponding wavelength absorption during the simulation. We apply this method to a nanosencond timescale molecular dynamics simulation of the light-harvesting peridinin-chlorophyll-protein complex from Amphidinium carterae, where chlorophyll was selected among the chromophores of the complex for the calculation. The combination of this quantum-classical calculation with the analysis of the large amplitude motions of the protein makes it possible to point out the relationship between the conformational flexibility of the environment and the excitation wavelength of the chromophore. Results support the idea of the existence of a correlation between protein conformational flexibility and chlorophyll electronic transitions induced by light. PMID- 12719216 TI - Modeling permeation energetics in the KcsA potassium channel. AB - The thermodynamics of cation permeation through the KcsA K(+) channel selectivity filter is studied from the perspective of a physically transparent semimicroscopic model using Monte Carlo free energy integration. The computational approach chosen permits dissection of the separate contributions to ionic stabilization arising from different parts of the channel (selectivity filter carbonyls, single-file water, cavity water, reaction field of bulk water, inner helices, ionizable residues). All features play important roles; their relative significance varies with the ion's position in the filter. The cavity appears to act as an electrostatic buffer, shielding filter ions from structural changes in the inner pore. The model exhibits K(+) vs. Na(+) selectivity, and roughly isoenergetic profiles for K(+) and Rb(+), and discriminates against Cs(+), all in agreement with experimental data. It also indicates that Ba(2+) and Na(+) compete effectively with permeant ions at a site near the boundary between the filter and the cavity, in the vicinity of the barium blocker site. PMID- 12719217 TI - Mechanically probing the folding pathway of single RNA molecules. AB - We study theoretically the denaturation of single RNA molecules by mechanical stretching, focusing on signatures of the (un)folding pathway in molecular fluctuations. Our model describes the interactions between nucleotides by incorporating the experimentally determined free energy rules for RNA secondary structure, whereas exterior single-stranded regions are modeled as freely jointed chains. For exemplary RNA sequences (hairpins and the Tetrahymena thermophila group I intron), we compute the quasiequilibrium fluctuations in the end-to-end distance as the molecule is unfolded by pulling on opposite ends. Unlike the average quasiequilibrium force-extension curves, these fluctuations reveal clear signatures from the unfolding of individual structural elements. We find that the resolution of these signatures depends on the spring constant of the force measuring device, with an optimal value intermediate between very rigid and very soft. We compare and relate our results to recent experiments by Liphardt et al. (2001). PMID- 12719218 TI - Feedback regulation in the lactose operon: a mathematical modeling study and comparison with experimental data. AB - A mathematical model for the regulation of induction in the lac operon in Escherichia coli is presented. This model takes into account the dynamics of the permease facilitating the internalization of external lactose; internal lactose; beta-galactosidase, which is involved in the conversion of lactose to allolactose, glucose and galactose; the allolactose interactions with the lac repressor; and mRNA. The final model consists of five nonlinear differential delay equations with delays due to the transcription and translation process. We have paid particular attention to the estimation of the parameters in the model. We have tested our model against two sets of beta-galactosidase activity versus time data, as well as a set of data on beta-galactosidase activity during periodic phosphate feeding. In all three cases we find excellent agreement between the data and the model predictions. Analytical and numerical studies also indicate that for physiologically realistic values of the external lactose and the bacterial growth rate, a regime exists where there may be bistable steady state behavior, and that this corresponds to a cusp bifurcation in the model dynamics. PMID- 12719219 TI - The Ca2+ dynamics of isolated mouse beta-cells and islets: implications for mathematical models. AB - [Ca(2+)](i) and electrical activity were compared in isolated beta-cells and islets using standard techniques. In islets, raising glucose caused a decrease in [Ca(2+)](i) followed by a plateau and then fast (2-3 min(-1)), slow (0.2-0.8 min( 1)), or a mixture of fast and slow [Ca(2+)](i) oscillations. In beta-cells, glucose transiently decreased and then increased [Ca(2+)](i), but no islet-like oscillations occurred. Simultaneous recordings of [Ca(2+)](i) and electrical activity suggested that differences in [Ca(2+)](i) signaling are due to differences in islet versus beta-cell electrical activity. Whereas islets exhibited bursts of spikes on medium/slow plateaus, isolated beta-cells were depolarized and exhibited spiking, fast-bursting, or spikeless plateaus. These electrical patterns in turn produced distinct [Ca(2+)](i) patterns. Thus, although isolated beta-cells display several key features of islets, their oscillations were faster and more irregular. beta-cells could display islet-like [Ca(2+)](i) oscillations if their electrical activity was converted to a slower islet-like pattern using dynamic clamp. Islet and beta-cell [Ca(2+)](i) changes followed membrane potential, suggesting that electrical activity is mainly responsible for the [Ca(2+)] dynamics of beta-cells and islets. A recent model consisting of two slow feedback processes and passive endoplasmic reticulum Ca(2+) release was able to account for islet [Ca(2+)](i) responses to glucose, islet oscillations, and conversion of single cell to islet-like [Ca(2+)](i) oscillations. With minimal parameter variation, the model could also account for the diverse behaviors of isolated beta-cells, suggesting that these behaviors reflect natural cell heterogeneity. These results support our recent model and point to the important role of beta-cell electrical events in controlling [Ca(2+)](i) over diverse time scales in islets. PMID- 12719220 TI - Role of the dielectric constants of membrane proteins and channel water in ion permeation. AB - Using both analytical solutions obtained from simplified systems and numerical results from more realistic cases, we investigate the role played by the dielectric constant of membrane proteins epsilon(p) and pore water epsilon(w) in permeation of ions across channels. We show that the boundary and its curvature are the crucial factors in determining how an ion's potential energy depends on the dielectric constants near an interface. The potential energy of an ion outside a globular protein has a dominant 1/epsilon(w) dependence, but this becomes 1/epsilon(p) for an ion inside a cavity. For channels, where the boundaries are in between these two extremes, the situation is more complex. In general, we find that variations in epsilon(w) have a much larger impact on the potential energy of an ion compared to those in epsilon(p). Therefore a better understanding of the effective epsilon(w) values employed in channel models is desirable. Although the precise value of epsilon(p) is not a crucial determinant of ion permeation properties, it still needs to be chosen carefully when quantitative comparisons with data are made. PMID- 12719221 TI - Profiling charge complementarity and selectivity for binding at the protein surface. AB - A novel analysis and representation of the protein surface in terms of electrostatic binding complementarity and selectivity is presented. The charge optimization methodology is applied in a probe-based approach that simulates the binding process to the target protein. The molecular surface is color coded according to calculated optimal charge or according to charge selectivity, i.e., the binding cost of deviating from the optimal charge. The optimal charge profile depends on both the protein shape and charge distribution whereas the charge selectivity profile depends only on protein shape. High selectivity is concentrated in well-shaped concave pockets, whereas solvent-exposed convex regions are not charge selective. This suggests the synergy of charge and shape selectivity hot spots toward molecular selection and recognition, as well as the asymmetry of charge selectivity at the binding interface of biomolecular systems. The charge complementarity and selectivity profiles map relevant electrostatic properties in a readily interpretable way and encode information that is quite different from that visualized in the standard electrostatic potential map of unbound proteins. PMID- 12719222 TI - Enhanced sampling of the molecular potential energy surface using mutually orthogonal latin squares: application to peptide structures. AB - The computational identification of the optimal three-dimensional fold of even a small peptide chain from its sequence, without reference to other known structures, is a complex problem. There have been several attempts at solving this by sampling the potential energy surface of the molecule in a systematic manner. Here we present a new method to carry out the sampling, and to identify low energy conformers of the molecule. The method uses mutually orthogonal Latin squares to select (of the order of) n(2) points from the multidimensional conformation space of size m(n), where n is the number of dimensions (i.e., the number of conformational variables), and m specifies the fineness of the search grid. The sampling is accomplished by first calculating the value of the potential energy function at each one of the selected points. This is followed by analysis of these values of the potential energy to obtain the optimal value for each of the n-variables separately. We show that the set of the n-optimal values obtained in this manner specifies a low energy conformation of the molecule. Repeated application of the method identifies other low energy structures. The computational complexity of this algorithm scales as the fourth power of the size of the molecule. We applied this method to several small peptides, such as the neuropeptide enkephalin, and could identify a set of low energy conformations for each. Many of the structures identified by this method have also been previously identified and characterized by experiment and theory. We also compared the best structures obtained for the tripeptide (Ala)(3) by the present method, with those obtained by an exhaustive grid search, and showed that the algorithm is successful in identifying all the low energy conformers of this molecule. PMID- 12719223 TI - Growth velocities of branched actin networks. AB - The growth of an actin network against an obstacle that stimulates branching locally is studied using several variants of a kinetic rate model based on the orientation-dependent number density of filaments. The model emphasizes the effects of branching and capping on the density of free filament ends. The variants differ in their treatment of side versus end branching and dimensionality, and assume that new branches are generated by existing branches (autocatalytic behavior) or independently of existing branches (nucleation behavior). In autocatalytic models, the network growth velocity is rigorously independent of the opposing force exerted by the obstacle, and the network density is proportional to the force. The dependence of the growth velocity on the branching and capping rates is evaluated by a numerical solution of the rate equations. In side-branching models, the growth velocity drops gradually to zero with decreasing branching rate, while in end-branching models the drop is abrupt. As the capping rate goes to zero, it is found that the behavior of the velocity is sensitive to the thickness of the branching region. Experiments are proposed for using these results to shed light on the nature of the branching process. PMID- 12719224 TI - Dynamics from a time series: can we extract the phase resetting curve from a time series? AB - Recordings of the membrane potential from a bursting neuron were used to reconstruct the phase curve for that neuron for a limited set of perturbations. These perturbations were inhibitory synaptic conductance pulses able to shift the membrane potential below the most hyperpolarized level attained in the free running mode. The extraction of the phase resetting curve from such a one dimensional time series requires reconstruction of the periodic activity in the form of a limit cycle attractor. Resetting was found to have two components. In the first component, if the pulse was applied during a burst, the burst was truncated, and the time until the next burst was shortened in a manner predicted by movement normal to the limit cycle. By movement normal to the limit cycle, we mean a switch between two well-defined solution branches of a relaxation-like oscillator in a hysteretic manner enabled by the existence of a singular dominant slow process (variable). In the second component, the onset of the burst was delayed until the end of the hyperpolarizing pulse. Thus, for the pulse amplitudes we studied, resetting was independent of amplitude but increased linearly with pulse duration. The predicted and the experimental phase resetting curves for a pyloric dilator neuron show satisfactory agreement. The method was applied to only one pulse per cycle, but our results suggest it could easily be generalized to accommodate multiple inputs. PMID- 12719225 TI - Sequence-function analysis of the K+-selective family of ion channels using a comprehensive alignment and the KcsA channel structure. AB - Sequence-function analysis of K(+)-selective channels was carried out in the context of the 3.2 A crystal structure of a K(+) channel (KcsA) from Streptomyces lividans (Doyle et al., 1998). The first step was the construction of an alignment of a comprehensive set of K(+)-selective channel sequences forming the putative permeation path. This pathway consists of two transmembrane segments plus an extracellular linker. Included in the alignment are channels from the eight major classes of K(+)-selective channels from a wide variety of species, displaying varied rectification, gating, and activation properties. Segments of the alignment were assigned to structural motifs based on the KcsA structure. The alignment's accuracy was verified by two observations on these motifs: 1), the most variability is shown in the turret region, which functionally is strongly implicated in susceptibility to toxin binding; and 2), the selectivity filter and pore helix are the most highly conserved regions. This alignment combined with the KcsA structure was used to assess whether clusters of contiguous residues linked by hydrophobic or electrostatic interactions in KcsA are conserved in the K(+)-selective channel family. Analysis of sequence conservation patterns in the alignment suggests that a cluster of conserved residues is critical for determining the degree of K(+) selectivity. The alignment also supports the near universality of the "glycine hinge" mechanism at the center of the inner helix for opening K channels. This mechanism has been suggested by the recent crystallization of a K channel in the open state. Further, the alignment reveals a second highly conserved glycine near the extracellular end of the inner helix, which may be important in minimizing deformation of the extracellular vestibule as the channel opens. These and other sequence-function relationships found in this analysis suggest that much of the permeation path architecture in KcsA is present in most K(+)-selective channels. Because of this finding, the alignment provides a robust starting point for homology modeling of the permeation paths of other K(+)-selective channel classes and elucidation of sequence-function relationships therein. To assay these applications, a homology model of the Shaker A channel permeation path was constructed using the alignment and KcsA as the template, and its structure evaluated in light of established structural criteria. PMID- 12719226 TI - Perfect and near-perfect adaptation in a model of bacterial chemotaxis. AB - The signaling apparatus mediating bacterial chemotaxis can adapt to a wide range of persistent external stimuli. In many cases, the bacterial activity returns to its prestimulus level exactly, and this perfect adaptability is robust against variations in various chemotaxis protein concentrations. We model the bacterial chemotaxis signaling pathway, from ligand binding to CheY phosphorylation. By solving the steady-state equations of the model analytically, we derive a full set of conditions for the system to achieve perfect adaptation. The conditions related to the phosphorylation part of the pathway are discovered for the first time, while other conditions are generalizations of the ones found in previous works. Sensitivity of the perfect adaptation is evaluated by perturbing these conditions. We find that, even in the absence of some of the perfect adaptation conditions, adaptation can be achieved with near-perfect precision as a result of the separation of scales in both chemotaxis protein concentrations and reaction rates, or specific properties of the receptor distribution in different methylation states. Since near-perfect adaptation can be found in much larger regions of the parameter space than that defined by the perfect adaptation conditions, their existence is essential to understand robustness in bacterial chemotaxis. PMID- 12719227 TI - Capsaicin activation of the pain receptor, VR1: multiple open states from both partial and full binding. AB - Capsaicin, the pungent ingredient of hot peppers, has long been used to identify nociceptors. Its molecular target, the vanilloid receptor VR1, was recently cloned and confirmed functionally as a polymodal detector of multiple pain stimuli: heat, acid, and vanilloids. Previous electrophysiology studies have focused on whole-cell characteristics of the receptor. Here, we provide the first in-depth single-channel kinetic study of VR1 to understand its activation mechanism. At low to medium concentrations, channel activity appeared as bursts. Not only did the durations of the interburst gaps vary with capsaicin, the bursts also appeared ligand-dependent, with high capsaicin prolonging bursts and stabilizing openings. Gating involved at least five closed and three open states, with strong correlations between short closures and long openings, and long closures and short openings. Increasing capsaicin reduced the long closures with little effect on short ones. The open time constants changed little with capsaicin concentration, though their relative proportions varied. These results suggest that 1), the channel contains multiple capsaicin binding sites; 2), both partial and full binding are capable of opening the channel; 3), when activated, multiple open states are accessible irrespective of the level of binding; and 4), capsaicin association occurs preferentially to the closed channel. PMID- 12719228 TI - Stimulatory action of internal protons on Slo1 BK channels. AB - We investigated the internal pH-sensitivity of heterologously expressed hSlo1 BK channels. In the virtual absence of Ca(2+) and Mg(2+) to isolate the voltage dependent gating transitions, low internal pH enhanced macroscopic hSlo1 currents by shifting the voltage-dependence of activation to more negative voltages. The activation time course was faster and the deactivation time course was slower with low pH. The estimated K(d) value of the stimulatory effect was approximately pH = 6.5 or 0.35 micro M. The stimulatory effect was maintained when the auxiliary subunit mouse beta1 was coexpressed. Treatment of the hSlo1 channel with the histidine modifying agent diethyl pyrocarbonate also enhanced the hSlo1 currents and greatly diminished the internal pH sensitivity, suggesting that diethyl pyrocarbonate and low pH may work on the same effector mechanism. High concentrations of Ca(2+) or Mg(2+) also masked the stimulatory effect of low internal pH. These results indicate that the acid-sensitivity of the Slo BK channel may involve the channel domain implicated in the divalent-dependent activation. PMID- 12719230 TI - On translocation through a membrane channel via an internal binding site: kinetics and voltage dependence. AB - Here we present a model for maltodextrin translocation through maltoporin channels. In a first step, our theoretical analysis does consider the case of a single binding site for a given substrate in a structurally unaffected channel with a possibly different entrance barrier on either side. It is shown how by means of conventional electrical conductance measurements (including current noise analysis) the basic equilibrium and rate constants can be determined as functions of the applied voltage. Then also the net translocation rate of the substrate becomes accessible quantitatively. This most simple model mechanism has been extended to include a voltage-dependent fast conformational change of the channel that prevents the binding process. The so developed approach has been tested with experimental data for a single maltoporin trimer being reconstituted in black lipid membranes when studied in the presence of maltohexaose as the substrate. The experimental results turned out to be clearly incompatible with binding alone. They are, however, very satisfactorily fitted by pertinent theoretical curves if also inhibition of binding by a conformational transition is taken into account. Accordingly, quantitative evaluations of the underlying parameters and eventually of the translocation rate have been carried out successfully. Our analysis reveals a set of parameters necessary for an optimal translocation that nicely corresponds to natural conditions. PMID- 12719229 TI - Comparison of the TIM and TOM channel activities of the mitochondrial protein import complexes. AB - Water-filled channels are central to the process of translocating proteins since they provide aqueous pathways through the hydrophobic environment of membranes. The Tom and Tim complexes translocate precursors across the mitochondrial outer and inner membranes, respectively, and contain channels referred to as TOM and TIM (previously called PSC and MCC). In this study, little differences were revealed from a direct comparison of the single channel properties of the TOM and TIM channels of yeast mitochondria. As they perform similar functions in translocating proteins across membranes, it is not surprising that both channels are high conductance, voltage-dependent channels that are slightly cation selective. Reconstituted TIM and TOM channel activities are not modified by deletion of the outer membrane channel VDAC, but are similarly affected by signal sequence peptides. PMID- 12719231 TI - Tonic and phasic guanidinium toxin-block of skeletal muscle Na channels expressed in Mammalian cells. AB - The blockage of skeletal muscle sodium channels by tetrodotoxin (TTX) and saxitoxin (STX) have been studied in CHO cells permanently expressing rat Nav1.4 channels. Tonic and use-dependent blockage were analyzed in the framework of the ion-trapped model. The tonic affinity (26.6 nM) and the maximum affinity (7.7 nM) of TTX, as well as the "on" and "off" rate constants measured in this preparation, are in remarkably good agreement with those measured for Nav1.2 expressed in frog oocytes, indicating that the structure of the toxin receptor of Nav1.4 and Nav1.2 channels are very similar and that the expression method does not have any influence on the pore properties of the sodium channel. The higher affinity of STX for the sodium channels (tonic and maximum affinity of 1.8 nM and 0.74 nM respectively) is explained as an increase on the "on" rate constant (approximately 0.03 s(-1) nM(-1)), compared to that of TTX (approximately 0.003 s(-1) nM(-1)), while the "off" rate constant is the same for both toxins (approximately 0.02 s(-1)). Estimations of the free-energy differences of the toxin-channel interaction indicate that STX is bound in a more external position than TTX. Similarly, the comparison of the toxins free energy of binding to a ion free, Na(+)- and Ca(2+)-occupied channel, is consistent with a binding site in the selectivity filter for Ca(2+) more external than for Na(+). This data may be useful in further attempts at sodium-channel pore modeling. PMID- 12719232 TI - Distinctive modulatory effects of five human auxiliary beta2 subunit splice variants on L-type calcium channel gating. AB - Sequence analysis of the human genome permitted cloning of five Ca(2+)-channel beta(2) splice variants (beta(2a)-beta(2e)) that differed only in their proximal amino-termini. The functional consequences of such beta(2)-subunit diversity were explored in recombinant L-type channels reconstituted in HEK 293 cells. Beta(2a) and beta(2e) targeted autonomously to the plasma membrane, whereas beta(2b) beta(2d) localized to the cytosol when expressed in HEK 293 cells. The pattern of modulation of L-type channel voltage-dependent inactivation gating correlated with the subcellular localization of the component beta(2) variant-membrane-bound beta(2a) and beta(2e) subunits conferred slow(er) channel inactivation kinetics and displayed a smaller fraction of channels recovering from inactivation with fast kinetics, compared to beta(2b)-beta(2d) channels. The varying effects of beta(2) subunits on inactivation gating were accounted for by a quantitative model in which L-type channels reversibly distributed between fast and slow forms of voltage-dependent inactivation-membrane-bound beta(2) subunits substantially decreased the steady-state fraction of fast inactivating channels. Finally, the beta(2) variants also had distinctive effects on L-type channel steady-state activation gating, as revealed by differences in the waveforms of tail-activation (G-V) curves, and conferred differing degrees of prepulse facilitation to the channel. Our results predict important physiological consequences arising from subtle changes in Ca(2+)-channel beta(2)-subunit structure due to alternative splicing and emphasize the utility of splice variants in probing structure function mechanisms. PMID- 12719233 TI - BeKm-1 is a HERG-specific toxin that shares the structure with ChTx but the mechanism of action with ErgTx1. AB - Peptide toxins with disulfide-stabilized structures have been used as molecular calipers to probe the outer vestibule structure of K channels. We want to apply this approach to the human ether-a-go-go-related gene (HERG) channel, whose outer vestibule is unique in structure and function among voltage-gated K channels. Our focus here is BeKm-1, a HERG-specific peptide toxin that can suppress HERG in the low nM concentration range. Although BeKm-1 shares the three-dimensional scaffold with the well-studied charybdotoxin, the two use different mechanisms in suppressing currents through their target K channels. BeKm-1 binds near, but not inside, the HERG pore, and it is possible that BeKm-1-bound HERG channels can conduct currents although with markedly altered voltage-dependence and kinetics of gating. BeKm-1 and ErgTx1 differ in three-dimensional scaffold, but the two share mechanism of action and have overlapping binding sites on the HERG channel. For both, residues in the middle of the S5-P linker (the putative 583-597 helix) and residues at the pore entrance are critical for binding, although specific contact points vary between the two. Toxin foot printing using BeKm-1 and ErgTx1 will likely provide complementary information about the unique outer vestibule structure of the HERG channel. PMID- 12719234 TI - Mechanism of membrane binding by the bovine seminal plasma protein, PDC-109: a surface plasmon resonance study. AB - PDC-109, the major protein of bovine seminal plasma, binds to sperm plasma membranes upon ejaculation and plays a crucial role in the subsequent events leading to fertilization. The binding process is mediated primarily by the specific interaction of PDC-109 with choline-containing phospholipids. In the present study the kinetics and mechanism of the interaction of PDC-109 with phospholipid membranes were investigated by the surface plasmon resonance technique. Binding of PDC-109 to different phospholipid membranes containing 20% cholesterol (wt/wt) indicated that binding occurs by a single-step mechanism. The association rate constant (k(1)) for the binding of PDC-109 to dimyristoylphosphatidylcholine (DMPC) membranes containing cholesterol was estimated to be 5.7 x 10(5) M(-1) s(-1) at 20 degrees C, while the values of k(1) estimated at the same temperature for the binding to membranes of negatively charged phospholipids such as dimyristoylphosphatidylglycerol (DMPG) and dimyristoylphosphatidic acid (DMPA) containing 20% cholesterol (wt/wt) were at least three orders of magnitude lower. The dissociation rate constant (k(-1)) for the DMPC/PDC-109 system was found to be 2.7 x 10(-2) s(-1) whereas the k(-1) values obtained with DMPG and DMPA was about three to four times higher. From the kinetic data, the association constant for the binding of PDC-109 to DMPC was estimated as 2.1 x 10(7) M(-1). The association constants for different phospholipids investigated decrease in the order: DMPC > DMPG > DMPA > DMPE. Thus the higher affinity of PDC-109 for choline phospholipids is reflected in a faster association rate constant and a slower dissociation rate constant for DMPC as compared to the other phospholipids. Binding of PDC-109 to dimyristoylphosphatidylethanolamine and dipalmitoylphosphatidylethanolamine, which are also zwitterionic, was found to be very weak, clearly indicating that the charge on the lipid headgroup is not the determining factor for the binding. Analysis of the activation parameters indicates that the interaction of PDC-109 with DMPC membranes is favored by a strong entropic contribution, whereas negative entropic contribution is primarily responsible for the rather weak interaction of this protein with DMPA and DMPG. PMID- 12719235 TI - Trehalose maintains phase separation in an air-dried binary lipid mixture. AB - Mixing and thermal behavior of hydrated and air-dried mixtures of 1,2-dilauroyl sn-glycero-3-phosphocholine (DLPC) and 1,2-distearoyl-d70-sn-glycero-3 phosphocholine (DSPCd-70) in the absence and presence of trehalose were investigated by Fourier transform infrared spectroscopy. Mixtures of DLPC:DSPCd 70 (1:1) that were air-dried at 25 degrees C show multiple phase transitions and mixed phases in the dry state. After annealing at high temperatures, however, only one transition is seen during cooling scans. When dried in the presence of trehalose, the DLPC component shows two phase transitions at -22 degrees C and 75 degrees C and is not fully solidified at -22 degrees C. The DSPCd-70 component, however, shows a single phase transition at 78 degrees C. The temperatures of these transitions are dramatically reduced after annealing at high temperatures with trehalose. The data suggest that the sugar has a fluidizing effect on the DLPC component during drying and that this effect becomes stronger for both components with heating. Examination of infrared bands arising from the lipid phosphate and sugar hydroxyl groups suggests that the strong effect of trehalose results from direct interactions between lipid headgroups and the sugar and that these interactions become stronger after heating. The findings are discussed in terms of the protective effect of trehalose on dry membranes. PMID- 12719236 TI - MSI-78, an analogue of the magainin antimicrobial peptides, disrupts lipid bilayer structure via positive curvature strain. AB - In this work, we present the first characterization of the cell lysing mechanism of MSI-78, an antimicrobial peptide. MSI-78 is an amphipathic alpha-helical peptide designed by Genaera Corporation as a synthetic analog to peptides from the magainin family. (31)P-NMR of mechanically aligned samples and differential scanning calorimetry (DSC) were used to study peptide-containing lipid bilayers. DSC showed that MSI-78 increased the fluid lamellar to inverted hexagonal phase transition temperature of 1,2-dipalmitoleoyl-phosphatidylethanolamine indicating the peptide induces positive curvature strain in lipid bilayers. (31)P-NMR of lipid bilayers composed of MSI-78 and 1-palmitoyl-2-oleoyl phosphatidylethanolamine demonstrated that the peptide inhibited the fluid lamellar to inverted hexagonal phase transition of 1-palmitoyl-2-oleoyl phosphatidylethanolamine, supporting the DSC results, and the peptide did not induce the formation of nonlamellar phases, even at very high peptide concentrations (15 mol %). (31)P-NMR of samples containing 1-palmitoyl-2-oleoyl phosphatidylcholine and MSI-78 revealed that MSI-78 induces significant changes in the bilayer structure, particularly at high peptide concentrations. At lower concentrations (1-5%), the peptide altered the morphology of the bilayer in a way consistent with the formation of a toroidal pore. Higher concentrations of peptide (10-15%) led to the formation of a mixture of normal hexagonal phase and lamellar phase lipids. This work shows that MSI-78 induces significant changes in lipid bilayers via positive curvature strain and presents a model consistent with both the observed spectral changes and previously published work. PMID- 12719237 TI - Cooperative partition model of nystatin interaction with phospholipid vesicles. AB - Nystatin is a membrane-active polyene antibiotic that is thought to kill fungal cells by forming ion-permeable channels. In this report we have investigated nystatin interaction with phosphatidylcholine liposomes of different sizes (large and small unilamellar vesicles) by time-resolved fluorescence measurements. Our data show that the fluorescence emission decay kinetics of the antibiotic interacting with gel-phase 1,2-dipalmitoyl-sn-glycero-3-phosphocholine vesicles is controlled by the mean number of membrane-bound antibiotic molecules per liposome, . The transition from a monomeric to an oligomeric state of the antibiotic, which is associated with a sharp increase in nystatin mean fluorescence lifetime from approximately 7-10 to 35 ns, begins to occur at a critical concentration of 10 nystatin molecules per lipid vesicle. To gain further information about the transverse location (degree of penetration) of the membrane-bound antibiotic molecules, the spin-labeled fatty acids (5- and 16 doxyl stearic acids) were used in depth-dependent fluorescence quenching experiments. The results obtained show that monomeric nystatin is anchored at the phospholipid/water interface and suggest that nystatin oligomerization is accompanied by its insertion into the membrane. Globally, the experimental data was quantitatively described by a cooperative partition model which assumes that monomeric nystatin molecules partition into the lipid bilayer surface and reversibly assemble into aggregates of 6 +/- 2 antibiotic molecules. PMID- 12719238 TI - The effect of cholesterol on the lateral diffusion of phospholipids in oriented bilayers. AB - Pulsed field gradient NMR was utilized to directly determine the lipid lateral diffusion coefficient for the following macroscopically aligned bilayers: dimyristoylphosphatidylcholine (DMPC), sphingomyelin (SM), palmitoyloleoylphosphatidylcholine (POPC), and dioleoylphosphatidylcholine (DOPC) with addition of cholesterol (CHOL) up to approximately 40 mol %. The observed effect of cholesterol on the lipid lateral diffusion is interpreted in terms of the different diffusion coefficients obtained in the liquid ordered (l(o)) and the liquid disordered (l(d)) phases occurring in the phase diagrams. Generally, the lipid lateral diffusion coefficient decreases linearly with increasing CHOL concentration in the l(d) phase for the PC-systems, while it is almost independent of CHOL for the SM-system. In this region the temperature dependence of the diffusion was always of the Arrhenius type with apparent activation energies (E(A)) in the range of 28-40 kJ/mol. The l(o) phase was characterized by smaller diffusion coefficients and weak or no dependence on the CHOL content. The E(A) for this phase was significantly larger (55-65 kJ/mol) than for the l(d) phase. The diffusion coefficients in the two-phase regions were compatible with a fast exchange between the l(d) and l(o) regions in the bilayer on the timescale of the NMR experiment (100 ms). Thus, strong evidence has been obtained that fluid domains (with size of micro m or less) with high molecular ordering are formed within a single lipid bilayer. These domains may play an important role for proteins involved in membrane functioning frequently discussed in the recent literature. The phase diagrams obtained from the analysis of the diffusion data are in qualitative agreement with earlier published ones for the SM/CHOL and DMPC/CHOL systems. For the DOPC/CHOL and the POPC/CHOL systems no two-phase behavior were observed, and the obtained E(A):s indicate that these systems are in the l(d) phase at all CHOL contents for temperatures above 25 degrees C. PMID- 12719239 TI - An experimental and theoretical analysis of ultrasound-induced permeabilization of cell membranes. AB - Application of ultrasound transiently permeabilizes cell membranes and offers a nonchemical, nonviral, and noninvasive method for cellular drug delivery. Although the ability of ultrasound to increase transmembrane transport has been well demonstrated, a systematic dependence of transport on ultrasound parameters is not known. This study examined cell viability and cellular uptake of calcein using 3T3 mouse cell suspension as a model system. Cells were exposed to varying acoustic energy doses at four different frequencies in the low frequency regime (20-100 kHz). At all frequencies, cell viability decreased with increasing acoustic energy dose, while the fraction of cells exhibiting uptake of calcein showed a maximum at an intermediate energy dose. Acoustic spectra under various ultrasound conditions were also collected and assessed for the magnitude of broadband noise and subharmonic peaks. While the cell viability and transport data did not show any correlation with subharmonic (f/2) emission, they correlated with the broadband noise, suggesting a dominant contribution of transient cavitation. A theoretical model was developed to relate reversible and irreversible membrane permeabilization to the number of transient cavitation events. The model showed that nearly every stage of transient cavitation, including bubble expansion, collapse, and subsequent shock waves may contribute to membrane permeabilization. For each mechanism, the volume around the bubble within which bubbles induce reversible and irreversible membrane permeabilization was determined. Predictions of the model are consistent with experimental data. PMID- 12719240 TI - Cholesterol in bilayers of sphingomyelin or dihydrosphingomyelin at concentrations found in ocular lens membranes. AB - Membranes of the lens of the eye of mammals have two particular characteristics, high concentrations of sphingomyelin, and dihydrosphingomyelin and cholesterol. We have studied the miscibility of cholesterol with both egg sphingomyelin and with dihydrosphingomyelin made by hydrogenation of egg sphingomyelin. At a cholesterol mol fraction of 0.5 and lower, crystallites of cholesterol are not present with either form of sphingomyelin, as observed by differential scanning calorimetry and by (13)C CP/MAS NMR. However, in the range of 0.6 to 0.8 mol fraction of cholesterol increasing amounts of crystallites form, with the amount of anhydrous cholesterol crystals formed being somewhat greater with dihyrosphingomyelin compared with sphingomyelin. Interestingly, cholesterol monohydrate crystallites formed in these two phospholipids exhibit a temperature of dehydration higher than that of pure cholesterol monohydrate crystals. These cholesterol monohydrate crystals form more rapidly and in greater amounts with the unmodified form of sphingomyelin. This difference is likely a consequence of differences at the membrane interface. The chemical shift of the (13)C of the carbonyl group, as measured by CP/MAS NMR, shows that there are differences between the two phospholipids in both the presence and absence of cholesterol. The bilayers with dihydrosphingomyelin are more hydrogen bonded. Cholesterol crystallites are known to be present in the lens of the eye. Our studies show that the ratio of sphingomyelin to dihydrosphingomyelin can affect the rate of formation of these cholesterol crystallites and thus play a role in the membrane of cells of the lens, affecting ocular function. PMID- 12719241 TI - Mechanism of the lamellar/inverse hexagonal phase transition examined by high resolution x-ray diffraction. AB - For the first time the electron density of the lamellar liquid crystalline as well as of the inverted hexagonal phase could be retrieved at the transition temperature. A reliable decomposition of the d-spacings into hydrophobic and hydrophilic structure elements could be performed owing to the presence of a sufficient number of reflections. While the hydrocarbon chain length, d(C), in the lamellar phase with a value of 14.5 A lies within the extreme limits of the estimated chain length of the inverse hexagonal phase 10 A < d(C) < 16 A, the changes in the hydrophilic region vary strongly. During the lamellar-to-inverse hexagonal phase transition the area per lipid molecule reduces by approximately 25%, and the number of water molecules per lipid increases from 14 to 18. On the basis of the analysis of the structural components of each phase, the interface between the coexisting mesophases between 66 and 84 degrees C has been examined in detail, and a model for the formation of the first rods in the matrix of the lamellar phospholipid stack is discussed. Judging from the structural relations between the inverse hexagonal and the lamellar phase, we suggest a cooperative chain reaction of rod formation at the transition midpoint, which is mainly driven by minimizing the interstitial region. PMID- 12719242 TI - Interaction between artificial membranes and enflurane, a general volatile anesthetic: DPPC-enflurane interaction. AB - The structural modifications of the dipalmitoylphosphatidylcholine (DPPC) organization induced by increasing concentration of the volatile anesthetic enflurane have been studied by differential scanning calorimetry, small-angle, and wide-angle x-ray scattering. The interaction of enflurane with DPPC depends on at least two factors: the enflurane-to-lipid concentration ratio and the initial organization of the lipids. At 25 degrees C (gel state), the penetration of enflurane within the lipids induces the apparition of two different mixed lipid phases. At low anesthetic-to-lipid molar ratio, the smectic distance increases whereas the direction of the chain tilt changes from a tilt toward next neighbors to a tilt between next-neighbors creating a new gel phase called L(beta')(2NNN). At high ratio, the smectic distance is much smaller than for the pure L(beta') DPPC phase, i.e., 50 A compared to 65 A, the aliphatic chains are perpendicular to the membrane and the fusion temperature of the phase is 33 degrees C. The electron profile of this phase that has been called L(beta)(i), indicates that the lipids are fully interdigitated. At 45 degrees C (fluid state), a new melted phase, called L(alpha)(2), was found, in which the smectic distance decreased compared to the initial pure L(alpha)(1) DPPC phase. The thermotropic behavior of the mixed phases has also been characterized by simultaneous x-ray scattering and differential scanning calorimetry measurements using the Microcalix calorimeter of our own. Finally, titration curves of enflurane effect in the mixed lipidic phase has been obtained by using the fluorescent lipid probe Laurdan. Measurements as a function of temperature or at constant temperature, i.e., 25 degrees C and 45 degrees C give, for the maximal effect, an enflurane-to-lipid ratio (M/M), within the membrane, of 1 and 2 for the L(alpha)(2) and the L(beta)(i) lamellar phase respectively. All the results taken together allowed to draw a pseudo-binary phase diagram of enflurane dipalmitoylphosphatidylcholine in excess water. PMID- 12719243 TI - A calorimetric study of binary mixtures of dihydrosphingomyelin and sterols, sphingomyelin, or phosphatidylcholine. AB - The thermotropic properties of binary mixtures of D-erythro-n-palmitoyl dihydrosphingomyelin (16:0-DHSM), D-erythro-n-palmitoyl-sphingomyelin (16:0-SM), cholesterol, lathosterol, and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) were studied by differential scanning calorimetry. Addition of sterol to 16:0 DHSM and 16:0-SM bilayers resulted in a progressive decrease in both the T(m) and the enthalpy of the main transition. The sterol-induced broad components in 16:0 DHSM endotherms had markedly lower enthalpies than those induced in 16:0-SM. Pretransitions recorded in 16:0-DHSM and 16:0-SM membranes responded differently to low concentrations of cholesterol. The presence of 5 mol % cholesterol increased the pretransition temperature in 16:0-SM bilayers, whereas it decreased the temperature in 16:0-DHSM membranes. Lathosterol behaved in general as cholesterol with regard to its effects on the thermotropic behavior of both sphingolipids, but it appeared to form more stable sterol-rich domains, as seen from the higher T(m) of the broad component, in comparison to cholesterol. Thermograms recorded on binary mixtures of 16:0-SM:16:0-DHSM and DPPC:16:0-DHSM showed that 16:0-SM mixed nearly ideally with 16:0-DHSM, whereas DPPC mixing was less ideal in a 16:0-DHSM membrane. In conclusion, we observed that 16:0-DHSM interactions with sterols differed from that seen with 16:0-SM, and that 16:0 DHSM mixed better with 16:0-SM than DPPC, which indicates that DHSM could function as a membrane organizer within laterally condensed domains. PMID- 12719244 TI - Structural requirements of the fructan-lipid interaction. AB - Fructans are a group of fructose-based oligo- and polysaccharides. They are proposed to be involved in membrane protection of plants during dehydration. In accordance with this hypothesis, they show an interaction with hydrated lipid model systems. However, the structural requirements for this interaction are not known both with respect to the fructans as to the lipids. To get insight into this matter, the interaction of several inulins and levan with lipids was investigated using a monomolecular lipid system or the MC 540 probe in a bilayer system. MD was used to get conformational information concerning the polysaccharides. It was found that levan-type fructan interacted comparably with model membranes composed of glyco- or phospholipids but showed a preference for lipids with a small headgroup. Furthermore, it was found that there was an inulin chain-length-dependent interaction with lipids. The results also suggested that inulin-type fructan had a more profound interaction with the membrane than levan type fructan. MD simulations indicated that the favorable conformation for levan is a helix, whereas inulin tends to form random coil structures. This suggests that flexibility is an important determinant for the fructan-lipid interaction. PMID- 12719245 TI - Cooperative regulation of myosin-actin interactions by a continuous flexible chain I: actin-tropomyosin systems. AB - We present a model for cooperative myosin binding to the regulated actin filament, where tropomyosins are treated as a weakly-confined continuous flexible chain covering myosin binding sites. Thermal fluctuations in chain orientation are initially required for myosin binding, leaving kinked regions under which subsequent myosins may bind without further distortion of the chain. Statistical mechanics predicts the fraction of sites with bound myosin-S1 as a function of their affinities. Published S1 binding curves to regulated filaments with different tropomyosin isoforms are fitted by varying the binding constant, chain persistence length nu (in actin monomers), and chain kink energy A from a single bound S1. With skeletal tropomyosin, we find an S1 actin-binding constant of 2.2 x 10(7) M(-1), A = 1.6 k(B)T and nu = 2.7. Similar persistence lengths are found with yeast tropomyosin. Larger values are found for tropomyosin-troponin in the presence of calcium (nu = 3.7) and tropomyosins from smooth muscle and fibroblasts (nu = 4.5). The relationship of these results to structural information and the rigid-unit model of McKillop and Geeves is discussed. PMID- 12719246 TI - Cooperative regulation of myosin-actin interactions by a continuous flexible chain II: actin-tropomyosin-troponin and regulation by calcium. AB - The model of myosin regulation by a continuous tropomyosin chain is generalized to a chain of tropomyosin-troponin units. Myosin binding to regulated actin is cooperative and initially inhibited by the chain as before. In the absence of calcium, myosin is further inhibited by the binding of troponin-I to actin, which through the whole of troponin pins the tropomyosin chain in a blocking position; myosin and TnI compete for actin and induce oppositely-directed chain kinks. The model predicts equilibrium binding curves for myosin-S1 and TnI as a function of their first-order affinities K(S1) and L(TI). Myosin is detached by the actin binding of TnI, but TnI is more efficiently detached by myosin when the kink size (typically nine to ten actin sites) spans the seven-site spacing between adjacent TnI molecules. An allosteric mechanism is used for coupling the detachment of TnI to calcium binding by TnC. With thermally activated TnI kinks (kink energy B approximately k(B)T), TnI also binds cooperatively to actin, producing cooperative detachment of myosin and biphasic myosin-calcium Hill plots, with Hill coefficients of 2 at high calcium and 4-6 at low calcium as observed in striated muscle. The theory also predicts the cooperative effects observed in the calcium loading of TnC. PMID- 12719247 TI - Tropomyosin ends determine the stability and functionality of overlap and troponin T complexes. AB - Tropomyosin binds end to end along the actin filament. Tropomyosin ends, and the complex they form, are required for actin binding, cooperative regulation of actin filaments by myosin, and binding to the regulatory protein, troponin T. The aim of the work was to understand the isoform and structural specificity of the end-to-end association of tropomyosin. The ability of N-terminal and C-terminal model peptides with sequences of alternate alpha-tropomyosin isoforms, and a troponin T fragment that binds to the tropomyosin overlap, to form complexes was analyzed using circular dichroism spectroscopy. Analysis of N-terminal extensions (N-acetylation, Gly, AlaSer) showed that to form an overlap complex between the N terminus and the C-terminus requires that the N-terminus be able to form a coiled coil. Formation of a ternary complex with the troponin T fragment, however, effectively takes place only when the overlap complex sequences are those found in striated muscle tropomyosins. Striated muscle tropomyosins with N-terminal modifications formed ternary complexes with troponin T that varied in affinity in the order: N-acetylated > Gly > AlaSer > unacetylated. The circular dichroism results were corroborated by native gel electrophoresis, and the ability of the troponin T fragment to promote binding of full-length tropomyosins to filamentous actin. PMID- 12719248 TI - EPR spectroscopy shows a microtubule-dependent conformational change in the kinesin switch 1 domain. AB - We have used site-directed spin-labeling and electron paramagnetic resonance spectroscopy to monitor a conformational change at the nucleotide site of kinesin. Cys-lite kinesin (K349 monomer) with the mutation S188C was spin labeled with MSL or MTSL. This residue is at the junction between the switch 1 region (which is a structure known to be sensitive to bound nucleotide in the G proteins) and the alpha3-helix, adjacent to the nucleotide site. The spectra showed two or more components of mobility, which were independent of nucleotide in the absence of microtubules (MTs). The spectra of both labels showed a change of mobility upon binding to MTs. A more mobile spectral component became enhanced for all triphosphate analogs examined, AMPPNP, ADP.AlFx, or ADP.BeFx, in the presence of MTs, although the magnitude of the new component and the degree of mobility varied with nucleotide analog. The ADP state showed a much-reduced spectral change with a small shift to the more immobilized component in the presence of MTs. For kinesin.ADP.MT, a van't Hoff plot gave DeltaH degrees = -96 kJ/mol implying that the conformational change was extensive. We conclude there is a conformational change in the switch 1-alpha3-helix domain when kinesin binds to MTs. PMID- 12719249 TI - Nucleosome repositioning via loop formation. AB - Active (catalyzed) and passive (intrinsic) nucleosome repositioning is known to be a crucial event during the transcriptional activation of certain eukaryotic genes. Here we consider theoretically the intrinsic mechanism and study in detail the energetics and dynamics of DNA-loop-mediated nucleosome repositioning, as previously proposed by earlier works. The surprising outcome of the present study is the inherent nonlocality of nucleosome motion within this model-being a direct physical consequence of the loop mechanism. On long enough DNA templates the longer jumps dominate over the previously predicted local motion, a fact that contrasts simple diffusive mechanisms considered before. The possible experimental outcome resulting from the considered mechanism is predicted, discussed, and compared to existing experimental findings. PMID- 12719250 TI - Single-strand stacking free energy from DNA beacon kinetics. AB - DNA beacons are short single-stranded chains which can form closed hairpin shapes through complementary base pairing at their ends. Contrary to the common polymer theory assumption that only their loop length matters, experiments show that their closing kinetics depend on the loop composition. We have modeled the closing kinetics and in so doing have obtained stacking enthalpies and entropies for single-stranded nucleic acids. The resulting change of persistence length with temperature effects the dynamics. With a Monte Carlo study, we answer another polymer question of how the closing time scales with chain length, finding tau approximately N(2.44+/-0.02). There is a significant crossover for shorter chains, bringing the effective exponent into good agreement with experiment. PMID- 12719251 TI - AC impedance spectroscopy of native DNA and M-DNA. AB - Monolayers of thiol-labeled DNA duplexes of 15, 20, and 30 basepairs were assembled on gold electrodes. Electron transfer was investigated by electrochemical impedance spectroscopy with Fe(CN)(6)(3-/4-) as a redox probe. The spectra, in the form of Nyquist plots, were analyzed with a modified Randles circuit which included an additional component in parallel, R(x), for the resistance through the DNA. For native B-DNA R(x) and R(ct), the charge transfer resistance, both increase with increasing length. M-DNA was formed by the addition of Zn(2+) at pH 8.6 and gave rise to characteristic changes in the Nyquist plots which were not observed upon addition of Mg(2+) or at pH 7.0. R(x) and R(ct) also increased with increasing duplex length for M-DNA but both were significantly lower compared to B-DNA. Therefore, electron transfer via the metal DNA film is faster than that of the native DNA film and certain metal ions can modulate the electrochemical properties of DNA monolayers. The results are consistent with an ion-assisted long-range polaron hopping mechanism for electron transfer. PMID- 12719252 TI - Stark spectroscopy on photoactive yellow protein, E46Q, and a nonisomerizing derivative, probes photo-induced charge motion. AB - The change in the electrostatic properties on excitation of the cofactor of wild type photoactive yellow protein (WT-PYP) have been directly determined using Stark-effect spectroscopy. We find that, instantaneously on photon absorption, there is a large change in the permanent dipole moment, /Delta(-->)mu/, (26 Debye) and in the polarizability, (-)Deltaalpha, (1000 A(3)). We expect such a large degree of charge motion to have a significant impact on the photocycle that is associated with the important blue-light negative phototactic response of Halorhodospira halophila. Furthermore, changing E46 to Q in WT-PYP does not significantly alter its electrostatic properties, whereas, altering the chromophore to prevent it from undergoing trans-cis isomerization results in a significant diminution of /Delta(-->)mu/ and (-)Deltaalpha. We propose that the enormous charge motion that occurs on excitation of 4-hydroxycinnamyl thioester, the chromophore in WT-PYP, plays a crucial role in initiating the photocycle by translocation of the negative charge, localized on the phenolate oxygen in the ground state, across the chromophore. We hypothesize that this charge motion would consequently increase the flexibility of the thioester tail thereby decreasing the activation barrier for the rotation of this moiety in the excited state. PMID- 12719253 TI - Energy trapping and detrapping in reaction center mutants from Rhodobacter sphaeroides. AB - Time-resolved fluorescence of chromatophores isolated from strains of Rhodobacter sphaeroides containing light harvesting complex I (LHI) and reaction center (RC) (no light harvesting complex II) was measured at several temperatures between 295 K and 10 K. Measurements were performed to investigate energy trapping from LHI to the RC in RC mutants that have a P/P(+) midpoint potential either above or below wild-type (WT). Six different strains were investigated: WT + LHI, four mutants with altered RC P/P(+) midpoint potentials, and an LHI-only strain. In the mutants with the highest P/P(+) midpoint potentials, the electron transfer rate decreases significantly, and at low temperatures it is possible to directly observe energy transfer from LHI to the RC by detecting the fluorescence kinetics from both complexes. In all mutants, fluorescence kinetics are multiexponential. To explain this, RC + LHI fluorescence kinetics were analyzed using target analysis in which specific kinetic models were compared. The kinetics at all temperatures can be well described with a model which accounts for the energy transfer between LHI and the RC and also includes the relaxation of the charge separated state P(+)H(A)(-), created in the RC as a result of the primary charge separation. PMID- 12719254 TI - The photochemical reaction cycle of proteorhodopsin at low pH. AB - The proton acceptor group in the recently described retinal protein, proteorhodopsin has an unusually high pK(a) of 7.1. It was shown that at pH above this pK(a), illumination initiates a photocycle similar to that of bacteriorhodopsin, and the protein transports proton across the cell membrane. Recently it was reported that proteorhodopsin, unlike bacteriorhodopsin, transports protons at pH below the pK(a) of the proton acceptor, and this transport is in the reverse direction. We have investigated the photocycle of proteorhodopsin at such low pH. At pH 5, three spectrally distinct intermediates K, L, and N, and another spectrally silent one, PR', could be identified, but a deprotonated Schiff base containing M-like intermediate, characteristic for proton pumping activity, does not accumulate. All the reactions between the intermediates are close to equilibrium, except the last transition from PR' to PR, when the protein returns to its initial unexcited state in a quasiunidirectional reaction. The electric signal measurements indicate that although charge motions are detected inside the protein, their net dislocation is zero, indicating that contrary to the earlier reported, at low pH no charged particle is transported across the membrane. PMID- 12719255 TI - Support vector machines for predicting membrane protein types by using functional domain composition. AB - Membrane proteins are generally classified into the following five types: 1), type I membrane protein; 2), type II membrane protein; 3), multipass transmembrane proteins; 4), lipid chain-anchored membrane proteins; and 5), GPI anchored membrane proteins. In this article, based on the concept of using the functional domain composition to define a protein, the Support Vector Machine algorithm is developed for predicting the membrane protein type. High success rates are obtained by both the self-consistency and jackknife tests. The current approach, complemented with the powerful covariant discriminant algorithm based on the pseudo-amino acid composition that has incorporated quasi-sequence-order effect as recently proposed by K. C. Chou (2001), may become a very useful high throughput tool in the area of bioinformatics and proteomics. PMID- 12719256 TI - Allosteric interactions within subsites of a monomeric enzyme: kinetics of fluorogenic substrates of PI-specific phospholipase C. AB - Two novel water-soluble fluorescein myo-inositol phosphate (FLIP) substrates, butyl-FLIP and methyl-FLIP, were used to examine the kinetics and subsite interactions of Bacillus cereus phosphatidylinositol-specific phospholipase C. Butyl-FLIP exhibited sigmoidal kinetics when initial rates are plotted versus substrate concentration. The data fit a Hill coefficient of 1.2-1.5, suggesting an allosteric interaction between two sites. Two substrate molecules bind to this enzyme, one at the active site and one at a subsite, causing an increase in activity. The kinetic behavior is mathematically similar to that of well-known cooperative multimeric enzymes even though this phosphatidylinositol-specific phospholipase C is a small, monomeric enzyme. The less hydrophobic substrate, methyl-FLIP, binds only to the active site and not the activator site, and thus exhibits standard hyperbolic kinetics. An analytical expression is presented that accounts for the kinetics of both substrates in the absence and presence of a nonsubstrate short-chain phospholipid, dihexanoylphosphatidylcholine. The fluorogenic substrates detect activation at much lower concentrations of dihexanoylphosphatidylcholine than previously reported. PMID- 12719257 TI - Molecular dynamics simulations on the first two helices of Vpu from HIV-1. AB - Vpu is an 81 amino acid protein of HIV-1 with two phosphorylation sites. It consists of a short N-terminal end traversing the bilayer and a longer cytoplasmic part. The dual functional role of Vpu is attributed to these topological distinct regions of the protein. The first 52 amino acids of Vpu (HV1H2) have been simulated, which are thought to be embedded in a fully hydrated lipid bilayer and to consist of a transmembrane helix (helix-1) connected via a flexible linker region, including a Glu-Tyr-Arg (EYR) motif, with a second helix (helix-2) residing with its helix long axis on the bilayer surface. Repeated molecular dynamics simulations show that Glu-28 is involved in salt bridge formation with Lys-31 and Arg-34 establishing a kink between the two helices. Helix-2 remains in a helical conformation indicating its stability and function as a "peptide float," separating helix-1 from the rest of the protein. This leads to the conclusion that Vpu consists of three functional modules: helix-1, helix 2, and the remaining residues toward the C-terminal end. PMID- 12719258 TI - Heme structural perturbation of PEG-modified horseradish peroxidase C in aromatic organic solvents probed by optical absorption and resonance Raman dispersion spectroscopy. AB - The heme structure perturbation of poly(ethylene glycol)-modified horseradish peroxidase (HRP-PEG) dissolved in benzene and toluene has been probed by resonance Raman dispersion spectroscopy. Analysis of the depolarization ratio dispersion of several Raman bands revealed an increase of rhombic B(1g) distortion with respect to native HRP in water. This finding strongly supports the notion that a solvent molecule has moved into the heme pocket where it stays in close proximity to one of the heme's pyrrole rings. The interactions between the solvent molecule, the heme, and the heme cavity slightly stabilize the hexacoordinate high spin state without eliminating the pentacoordinate quantum mixed spin state that is dominant in the resting enzyme. On the contrary, the model substrate benzohydroxamic acid strongly favors the hexacoordinate quantum mixed spin state and induces a B(2g)-type distortion owing to its position close to one of the heme methine bridges. These results strongly suggest that substrate binding must have an influence on the heme geometry of HRP and that the heme structure of the enzyme-substrate complex (as opposed to the resting state) must be the key to understanding the chemical reactivity of HRP. PMID- 12719259 TI - Rationalization of membrane protein crystallization with polyethylene glycol using a simple depletion model. AB - Based on the importance of crystallizing membrane proteins in a rational way, cytochrome bc(1) complex (BC1) was crystallized using polyethylene glycol (PEG) as a sole crystallization agent. Interaction between protein-detergent complexes of BC1 was estimated by dynamic light scattering, and was compared with the numerical calculation using the Derjaguin-Landau-Verwey-Overbeek potential plus a depletion potential, without considering specific surface properties of the protein-detergent complexes. The experiments and calculation were found to be consistent and we obtained a relation between PEG molecular weight M and the range of depletion zone delta as delta approximately M(0.48+/-0.02). The stability of liquid phase of BC1 solutions was controlled by a ratio of (the range of depletion zone)/(the radius of a BC1 particle), which was consistent with recent theoretical predictions. The crystallization was most successful under a condition where the stability of the liquid phase changed from stable to unstable. The PEG molecular weight that fulfilled this condition coincided with the one used empirically to crystallize BC1 in the past by a number of groups. These results are compared to the fact that membrane proteins were often successfully crystallized close to the detergent cloud point. PMID- 12719260 TI - Three-dimensional imaging of lipid gene-carriers: membrane charge density controls universal transfection behavior in lamellar cationic liposome-DNA complexes. AB - Cationic liposomes (CLs) are used worldwide as gene vectors (carriers) in nonviral clinical applications of gene delivery, albeit with unacceptably low transfection efficiencies (TE). We present three-dimensional laser scanning confocal microscopy studies revealing distinct interactions between CL-DNA complexes, for both lamellar L(alpha)(C) and inverted hexagonal H(II)(C) nanostructures, and mouse fibroblast cells. Confocal images of L(alpha)(C) complexes in cells identified two regimes. For low membrane charge density (sigma(M)), DNA remained trapped in CL-vectors. By contrast, for high sigma(M), released DNA was observed in the cytoplasm, indicative of escape from endosomes through fusion. Remarkably, firefly luciferase reporter gene studies in the highly complex L(alpha)(C)-mammalian cell system revealed an unexpected simplicity where, at a constant cationic to anionic charge ratio, TE data for univalent and multivalent cationic lipids merged into a single curve as a function of sigma(M), identifying it as a key universal parameter. The universal curve for transfection by L(alpha)(C) complexes climbs exponentially over approximately four decades with increasing sigma(M) below an optimal charge density (sigma(M)(*)), and saturates for at a value rivaling the high transfection efficiency of H(II)(C) complexes. In contrast, the transfection efficiency of H(II)(C) complexes is independent of sigma(M). The exponential dependence of TE on sigma(M) for L(alpha)(C) complexes, suggests the existence of a kinetic barrier against endosomal fusion, where an increase in sigma(M) lowers the barrier. In the saturated TE regime, for both L(alpha)(C) complexes and H(II)(C), confocal microscopy reveals the dissociation of lipid and DNA. However, the lipid-released DNA is observed to be in a condensed state, most likely with oppositely charged macro-ion condensing agents from the cytoplasm, which remain to be identified. Much of the observed bulk of condensed DNA may be transcriptionally inactive and may determine the current limiting factor to transfection by cationic lipid gene vectors. PMID- 12719261 TI - Energetic and structural considerations for the mechanism of protein sliding along DNA in the nonspecific BamHI-DNA complex. AB - The molecular mechanism by which DNA-binding proteins find their specific binding sites is still unclear. To gain insights into structural and energetic elements of this mechanism, we used the crystal structure of the nonspecific BamHI-DNA complex as a template to study the dominant electrostatic interaction in the nonspecific association of protein with DNA, and the possible sliding pathways that could be sustained by such an interaction. Based on calculations using the nonlinear Poisson-Boltzmann method and Brownian dynamics, a model is proposed for the initial nonspecific binding of BamHI to B-form DNA that differs from that seen in the crystal structure of the nonspecific complex. The model is electrostatically favorable, and the salt dependence as well as other thermodynamic parameters calculated for this model are in good agreement with experimental results. Several residues in BamHI are identified for their important contribution to the energy in the nonspecific binding model, and specific mutagenesis experiments are proposed to test the model on this basis. We show that a favorable sliding pathway of the protein along DNA is helical. PMID- 12719262 TI - Site-specific identification of non-beta-strand conformations in Alzheimer's beta amyloid fibrils by solid-state NMR. AB - The most well-established structural feature of amyloid fibrils is the cross-beta motif, an extended beta-sheet structure formed by beta-strands oriented perpendicular to the long fibril axis. Direct experimental identification of non beta-strand conformations in amyloid fibrils has not been reported previously. Here we report the results of solid-state NMR measurements on amyloid fibrils formed by the 40-residue beta-amyloid peptide associated with Alzheimer's disease (Abeta(1-40)), prepared synthetically with pairs of (13)C labels at consecutive backbone carbonyl sites. The measurements probe the peptide backbone conformation in residues 24-30, a segment where a non-beta-strand conformation has been suggested by earlier sequence analysis, cross-linking experiments, and molecular modeling. Data obtained with the fpRFDR-CT, DQCSA, and 2D MAS exchange solid state NMR techniques, which provide independent constraints on the phi and psi backbone torsion angles between the labeled carbonyl sites, indicate non-beta strand conformations at G25, S26, and G29. These results represent the first site specific identification and characterization of non-beta-strand peptide conformations in an amyloid fibril. PMID- 12719263 TI - Computational analysis of F-actin turnover in cortical actin meshworks using fluorescent speckle microscopy. AB - Fluorescent speckle microscopy (FSM) is a new imaging technique with the potential for simultaneous visualization of translocation and dynamic turnover of polymer structures. However, the use of FSM has been limited by the lack of specialized software for analysis of the positional and photometric fluctuations of hundreds of thousand speckles in an FSM time-lapse series, and for translating this data into biologically relevant information. In this paper we present a first version of a software for automated analysis of FSM movies. We focus on mapping the assembly and disassembly kinetics of a polymer meshwork. As a model system we have employed cortical F-actin meshworks in live newt lung epithelial cells. We lay out the algorithm in detail and present results of our analysis. The high spatial and temporal resolution of our maps reveals a kinetic cycling of F-actin, where phases of polymerization alternate with depolymerization in a spatially coordinated fashion. The cycle rates change when treating cells with a low dose of the drug latrunculin A. This shows the potential of this technique for future quantitative screening of drugs affecting the actin cytoskeleton. Various control experiments demonstrate that the algorithm is robust with respect to intensity variations due to noise and photobleaching and that effects of focus plane drifts can be eliminated by manual refocusing during image acquisition. PMID- 12719264 TI - Analyzing intracellular binding and diffusion with continuous fluorescence photobleaching. AB - Transport and binding of molecules to specific sites are necessary for the assembly and function of ordered supramolecular structures in cells. For analyzing these processes in vivo, we have developed a confocal fluorescence fluctuation microscope that allows both imaging of the spatial distribution of fluorescent molecules with confocal laser scanning microscopy and probing their mobility at specific positions in the cell with fluorescence correlation spectroscopy and continuous fluorescence photobleaching (CP). Because fluorescence correlation spectroscopy is restricted to rapidly diffusing particles and CP to slower processes, these two methods complement each other. For the analysis of binding-related contributions to mobility we have derived analytical expressions for the temporal behavior of CP curves from which the bound fraction and/or the dissociation rate or residence time at binding sites, respectively, can be obtained. In experiments, we investigated HeLa cells expressing different fluorescent proteins: Although enhanced green fluorescent protein (EGFP) shows high mobility, fusions of histone H2B with the yellow fluorescent protein are incorporated into chromatin, and these nuclei exhibit the presence of a stably bound and a freely diffusing species. Nonpermanent binding was found for mTTF-I, a transcription termination factor for RNA polymerase I, fused with EGFP. The cells show fluorescent nucleoli, and binding is transient. CP yields residence times for mTTF-I-EGFP of approximately 13 s. PMID- 12719265 TI - Lipid-gramicidin interactions: dynamic structure of the boundary lipid by 2D ELDOR. AB - The use of 2D-electron-electron double resonance (2D-ELDOR) for the characterization of the boundary lipid in membrane vesicles of DPPC and gramicidin A' (GA) is reported. We show that 2D-ELDOR, with its enhanced spectral resolution to dynamic structure as compared with continuous-wave electron spin resonance, provides a reliable and useful way of studying lipid-protein interactions. The 2D-ELDOR spectra of the end-chain spin label 16-PC in DPPC/GA vesicles is composed of two components, which are assigned to the bulk lipids (with sharp auto peaks and crosspeaks) and to the boundary lipids (with broad auto peaks). Their distinction is clearest for higher temperatures and higher GA concentrations. The quantitative analysis of these spectra shows relatively faster motions and very low ordering for the end chain of the bulk lipids, whereas the boundary lipids show very high "y-ordering" and slower motions. The y ordering represents a dynamic bending at the end of the boundary lipid acyl chain, which can then coat the GA molecules. These results are consistent with the previous studies by Ge and Freed (1999) using continuous-wave electron spin resonance, thereby supporting their model for GA aggregation and H(II) phase formation for high GA concentrations. Improved instrumental and simulation methods have been employed. PMID- 12719266 TI - Investigations into the life cycle of the bacterial predator Bdellovibrio bacteriovorus 109J at an interface by atomic force microscopy. AB - Atomic force microscopy was used to image Bdellovibrio bacteriovorus 109J, a gram negative bacterial predator that consumes a variety of other gram-negative bacteria. In predator-prey communities grown on filters at hydrated air-solid interfaces, repeated cycles of hunting, invasion, growth, and lysis occurred readily even though the cells were limited to near two-dimensional movement. This system allowed us to image the bacteria directly without extensive preparation or modification, and many of the cells remained alive during imaging. Presented are images of the life cycle in two species of prey organisms, both Escherichia coli (a small prey bacterium that grows two-dimensionally on a surface) and Aquaspirillum serpens (a large prey bacterium that grows three-dimensionally on a surface), including high-resolution images of invaded prey cells called bdelloplasts. We obtained evidence for multiple invasions per prey cell, as well as significant heterogeneity in morphology of bdellovibrios. Mutant host independent bdellovibrios were observed to have flagella and to excrete a coating that causes the predators to clump together on a surface. Most interestingly, changes in the texture of the cell surface membranes were measured during the course of the invasion cycle. Thus, coupled with our preparation method, atomic force microscopy allowed new observations to be made about Bdellovibrio at an interface. These studies raise important questions about the ways in which bacterial predation at interfaces (air-solid or liquid-solid) may be similar to or different from predation in solution. PMID- 12719267 TI - The mechanics of neutrophils: synthetic modeling of three experiments. AB - Much experimental data exist on the mechanical properties of neutrophils, but so far, they have mostly been approached within the framework of liquid droplet models. This has two main drawbacks: 1), It treats the cytoplasm as a single phase when in reality, it is a composite of cytosol and cytoskeleton; and 2), It does not address the problem of active neutrophil deformation and force generation. To fill these lacunae, we develop here a comprehensive continuum mechanical paradigm of the neutrophil that includes proper treatment of the membrane, cytosol, and cytoskeleton components. We further introduce two models of active force production: a cytoskeletal swelling force and a polymerization force. Armed with these tools, we present computer simulations of three classic experiments: the passive aspiration of a neutrophil into a micropipette, the active extension of a pseudopod by a neutrophil exposed to a local stimulus, and the crawling of a neutrophil inside a micropipette toward a chemoattractant against a varying counterpressure. Principal results include: 1), Membrane cortical tension is a global property of the neutrophil that is affected by local area-increasing shape changes. We argue that there exists an area dilation viscosity caused by the work of unfurling membrane-storing wrinkles and that this viscosity is responsible for much of the regulation of neutrophil deformation. 2), If there is no swelling force of the cytoskeleton, then it must be endowed with a strong cohesive elasticity to prevent phase separation from the cytosol during vigorous suction into a capillary tube. 3), We find that both swelling and polymerization force models are able to provide a unifying fit to the experimental data for the three experiments. However, force production required in the polymerization model is beyond what is expected from a simple short-range Brownian ratchet model. 4), It appears that, in the crawling of neutrophils or other amoeboid cells inside a micropipette, measurement of velocity versus counterpressure curves could provide a determination of whether cytoskeleton-to cytoskeleton interactions (such as swelling) or cytoskeleton-to-membrane interactions (such as polymerization force) are predominantly responsible for active protrusion. PMID- 12719268 TI - Analysis of cell kinetics using a cell division marker: mathematical modeling of experimental data. AB - We consider an age-maturity structured model arising from a blood cell proliferation problem. This model is "hybrid", i.e., continuous in time and age but the maturity variable is discrete. This is due to the fact that we include the cell division marker carboxyfluorescein diacetate succinimidyl ester. We use our mathematical analysis in conjunction with experimental data taken from the division analysis of primitive murine bone marrow cells to characterize the maturation/proliferation process. Cell cycle parameters such as proliferative rate beta, cell cycle duration tau, apoptosis rate gamma, and loss rate micro can be evaluated from CarboxyFluorescein diacetate Succinimidyl Ester + cell tracking experiments. Our results indicate that after three days in vitro, primitive murine bone marrow cells have parameters beta = 2.2 day(-1), tau = 0.3 day, gamma = 0.3 day(-1), and micro = 0.05 day(-1). PMID- 12719269 TI - Contribution of cyclic-nucleotide-gated channels to the resting conductance of olfactory receptor neurons. AB - The basal conductance of unstimulated frog olfactory receptor neurons was investigated using whole-cell and perforated-patch recording. The input conductance, measured between -80 mV and -60 mV, averaged 0.25 nS in physiological saline. Studies were conducted to determine whether part of the input conductance is due to gating of neuronal cyclic-nucleotide-gated (CNG) channels. In support of this idea, the neuronal resting conductance was reduced by each of five treatments that reduce current through CNG channels: external application of divalent cations or amiloride; treatment with either of two adenylate cyclase inhibitors; and application of AMP-PNP, a competitive substrate for adenylate cyclase. The current blocked by divalent cations or by a cyclase inhibitor reversed near 0 mV, as expected for a CNG current. Under physiological conditions, gating of CNG channels contributes approximately 0.06 nS to the resting neuronal conductance. This implies a resting cAMP concentration of 0.1 0.3 micro M. A theoretical model suggests that a neuron containing 0.1-0.3 micro M cAMP is poised to give the largest possible depolarization in response to a very small olfactory stimulus. Although having CNG channels open at rest decreases the voltage change resulting from a given receptor current, it more substantially increases the receptor current resulting from a given increase in [cAMP]. PMID- 12719270 TI - Heterogeneity of ADP diffusion and regulation of respiration in cardiac cells. AB - Heterogeneity of ADP diffusion and regulation of respiration were studied in permeabilized cardiomyocytes and cardiac fibers in situ and in silico. Regular arrangement of mitochondria in cells was altered by short-time treatment with trypsin and visualized by confocal microscopy. Manipulation of matrix volumes by changing K(+) and sucrose concentrations did not affect the affinity for ADP either in isolated heart mitochondria or in skinned fibers. Pyruvate kinase (PK) phosphoenolpyruvate (PEP) were used to trap ADP generated in Ca,MgATPase reactions. Inhibition of respiration by PK-PEP increased 2-3 times after disorganization of regular mitochondrial arrangement in cells. ADP produced locally in the mitochondrial creatine kinase reaction was not accessible to PK PEP in intact permeabilized fibers, but some part of it was released from mitochondria after short proteolysis due to increased permeability of outer mitochondrial membrane. In in silico studies we show by mathematical modeling that these results can be explained by heterogeneity of ADP diffusion due to its restrictions at the outer mitochondrial membrane and in close areas, which is changed after proteolysis. Localized restrictions and heterogeneity of ADP diffusion demonstrate the importance of mitochondrial functional complexes with sarcoplasmic reticulum and myofibrillar structures and creatine kinase in regulation of oxidative phosphorylation. PMID- 12719271 TI - Effects of permeant ion concentrations on the gating of L-type Ca2+ channels in hair cells. AB - We determined the gating and permeation properties of single L-type Ca(2+) channels, using hair cells and varying concentrations (5-70 mM) of the charge carriers Ba(2+) and Ca(2+). The channels showed distinct gating modes with high- and low-open probability. The half-activation voltage (V(1/2)) shifted in the hyperpolarizing direction from high to low permeant ion concentrations consistent with charge screening effects. However, the differences in the slope of the voltage shifts (in VM(-1)) between Ca(2+) (0.23) and Ba(2+) (0.13), suggest that channel-ion interaction may also contribute to the gating of the channel. We examined the effect of mixtures of Ba(2+) and Ca(2+) on the activation curve. In 5 mM Ca(2+), the V(1/2) was, -26.4 +/- 2.0 mV compared to Ba(2+), -34.7 +/- 2.9 mV, as the charge carrier. However, addition of 1 mM Ba(2+) in 4 mM Ca(2+), a molar ratio, which yielded an anomalous-mole fraction effect, was sufficient to shift the V(1/2) to -34.7 +/- 1.5 mV. Although Ca(2+)-dependent inactivation of the L-type channels in hair cells can yield the present findings, we provide evidence that the anomalous gating of the channel may stem from the closed interaction between ion permeation and gating. PMID- 12719272 TI - Effects of elevated extracellular potassium on the stimulation mechanism of diastolic cardiac tissue. AB - During cardiac disturbances such as ischemia and hyperkalemia, the extracellular potassium ion concentration is elevated. This in turn changes the resting transmembrane potential and affects the excitability of cardiac tissue. To test the hypothesis that extracellular potassium elevation also alters the stimulation mechanism, we used optical fluorescence imaging to examine the mechanism of diastolic anodal unipolar stimulation of cardiac tissue under 4 mM (normal) and 8 mM (elevated) extracellular potassium. We present several visualization methods that are useful for distinguishing between anodal-make and anodal-break excitation. In the 4-mM situation, stimulation occurred by the make, or stimulus onset, mechanism that involved propagation out of the virtual cathodes. For 8-mM extracellular potassium, the break or stimulus termination mechanism occurred with propagation out of the virtual anode. We conclude that elevated potassium, as might occur in myocardial ischemia, alters not only stimulation threshold but also the excitation mechanism for anodal stimulation. PMID- 12719273 TI - Micelle formation by a fragment of human islet amyloid polypeptide. AB - Human islet amyloid polypeptide (hIAPP) is the major component of amyloid plaques found in the pancreatic islets of persons with type 2 diabetes mellitus. HIAPP belongs to the group of amyloidogenic proteins, characterized by their aggregation and deposition as fibrillar amyloid in various body tissues. The aggregation of amyloidogenic proteins is thought to occur via a common pathway, but currently no unifying kinetic model exists. In previous work, we presented a model of amyloid fibril formation formulated from our observations of the aggregation of an amyloidogenic fragment of hIAPP, amino acids 20-29. Our model is based on nucleation-dependent aggregation, modified by the formation of off pathway hIAPP micelles. In the present study we confirm the presence of peptide micelles, and experimentally determine the critical micelle concentration in solutions of hIAPP fragments using three different techniques: conductivity, pH, and fluorescence. All three techniques yield a critical micelle concentration of 3-3.5 micro M peptide. Furthermore, based on changes in the fluorescence intensity of a labeled peptide fragment as well as a decrease in solution pH as a result of deprotonation of the amino terminus, we conclude that the amino terminus of the fragment undergoes a significant change of environment upon micellization. PMID- 12719275 TI - Rosuvastatin decreases caveolin-1 and improves nitric oxide-dependent heart rate and blood pressure variability in apolipoprotein E-/- mice in vivo. AB - BACKGROUND: Decreased heart rate variability (HRV) and increased blood pressure variability (BPV), determined in part by nitric oxide (NO)-dependent endothelial dysfunction, are correlated with adverse prognosis in cardiovascular diseases. We examined potential alterations in BPV and HRV in genetically dyslipidemic, apolipoprotein (apo) E-/-, and control mice and the effect of chronic statin treatment on these parameters in relation to their NO synthase (NOS)-modifying properties. METHODS AND RESULTS: BP and HR were recorded in unrestrained, nonanesthetized mice with implanted telemetry devices with or without rosuvastatin. Cardiac and aortic expression of endothelial NOS and caveolin-1 were measured by immunoblotting. Both systolic BP and HR were elevated in apoE-/- mice, with abolition of their circadian cycles. Spectral analysis showed an increase in their systolic BPV in the very-low-frequency (+17%) band and a decrease in HRV in the high-frequency (-57%) band, reflecting neurohumoral and autonomic dysfunction. Decreased sensitivity to acute injection of atropine or an NOS inhibitor indicated basal alterations in both parasympathetic and NOS regulatory systems in apoE-/- mice. Aortic caveolin-1 protein, an inhibitor of endothelial NOS, was also increased in these mice by 2.0-fold and correlated positively with systolic BPV in the very-low-frequency band. Rosuvastatin treatment corrected the hemodynamic and caveolin-1 expression changes despite persisting elevated plasma cholesterol levels. CONCLUSIONS: Rosuvastatin decreases caveolin-1 expression and promotes NOS function in apoE-/-, dyslipidemic mice in vivo, with concurrent improvements in BPV and HRV. This highlights the beneficial effects of rosuvastatin on cardiovascular function beyond those attributed to lipid lowering. PMID- 12719276 TI - Serum total cholesterol concentrations and awareness, treatment, and control of hypercholesterolemia among US adults: findings from the National Health and Nutrition Examination Survey, 1999 to 2000. AB - BACKGROUND: Serum cholesterol concentrations have decreased in the US population. Whether the decline continued during the 1990s is unknown. METHODS AND RESULTS: We used data from 4148 men and women aged > or =20 years who had a total cholesterol determination or reported using cholesterol-lowering medications and who participated in the National Health and Nutrition Examination Survey (NHANES) from 1999 to 2000 (this is a cross-sectional health examination survey of the US population), and we compared the results with data from 15 719 participants in NHANES III (1988 to 1994). For all adults, the age-adjusted mean total cholesterol concentration decreased from 5.31 mmol/L (205 mg/dL) in NHANES III to 5.27 mmol/L (203 mg/dL) in NHANES 1999 to 2000 (P=0.159). The age-adjusted mean total cholesterol concentration decreased by 0.02 mmol/L (0.7 mg/dL) among men (P=0.605) and 0.06 mmol/L (2.3 mg/dL) among women (P=0.130). Significant decreases were observed among men aged > or =75 years, black men, and Mexican American women. Among participants who had a total cholesterol concentration > or =5.2 mmol/L (200 mg/dL) or who reported using cholesterol-lowering medications, 69.5% reported having had their cholesterol checked, 35.0% were aware that they had hypercholesterolemia, 12.0% were on treatment, and 5.4% had a total cholesterol concentration <5.2 mmol/L (200 mg/dL) after age adjustment. CONCLUSIONS: The mean serum total cholesterol concentration of the adult US population in 1999 to 2000 has changed little since 1988 to 1994. The low percentage of adults with controlled blood cholesterol concentration suggests the need for a renewed commitment to the prevention, treatment, and control of hypercholesterolemia. PMID- 12719277 TI - Physical activity in relation to cardiovascular disease and total mortality among men with type 2 diabetes. AB - BACKGROUND: The present study was conducted to examine the relationship of physical activity with risk of cardiovascular disease (CVD) and mortality among men with type 2 diabetes. CVD risk and mortality are increased in type 2 diabetes. Few epidemiological studies have investigated the effect of physical activity on these outcomes among type 2 diabetics. METHODS AND RESULTS: Of the 3058 men who reported a diagnosis of diabetes at age 30 years or older in the Health Professionals' Follow-up Study (HPFS), we excluded 255 who reported a physical impairment. In the remaining 2803 men, physical activity was assessed every 2 years; 266 new cases of CVD and 355 deaths of all causes were identified during 14 years of follow-up. Relative risks of CVD and death were estimated from Cox proportional hazards analysis with adjustment for potential confounders. The multivariate relative risks of CVD incidence corresponding to quintiles of total physical activity were 1.0, 0.87, 0.64, 0.72, and 0.67 (Ptrend=0.07). The corresponding multivariate relative risks for total mortality were 1.0, 0.80, 0.57, 0.58, and 0.58 (Ptrend=0.005). Walking was associated with reduced risk of total mortality. Relative risks across quintiles of walking were 1.0, 0.97, 0.87, 0.97, and 0.57 (Ptrend=0.002). Walking pace was inversely associated with CVD, fatal CVD, and total mortality independently of walking hours. CONCLUSIONS: Physical activity was associated with reduced risk of CVD, cardiovascular death, and total mortality in men with type 2 diabetes. Walking and walking pace were associated with reduced total mortality. PMID- 12719278 TI - Promoter (4G/5G) plasminogen activator inhibitor-1 genotype and plasminogen activator inhibitor-1 levels in blacks, Hispanics, and non-Hispanic whites: the Insulin Resistance Atherosclerosis Study. AB - BACKGROUND: The 4G/5G polymorphism of the plasminogen activator inhibitor-1 (PAI 1) gene has been related to cardiovascular disease. METHODS AND RESULTS: Insulin resistance was measured with a frequently sampled intravenous glucose tolerance test in the Insulin Resistance Atherosclerosis Study (IRAS), and PAI-1 4G/5G promoter genotype was established by allele-specific polymerase chain reaction amplification of genomic DNA. There were 287 subjects with the 4G/4G genotype (18.4%), 691 heterozygote subjects (44.2%), and 586 carriers of the 5G/5G genotype (37.5%). The genotype distribution was different across the 3 ethnic groups (P=0.001). PAI-1 levels were lower in blacks than in non-Hispanic whites and Hispanics and lower in non-Hispanic whites than in Hispanics (all P=0.0001). Subjects homozygous for the 4G allele had the highest plasma PAI-1, heterozygote subjects were intermediate, and 5G homozygotes had the lowest levels of PAI-1. These patterns remained unaffected by adjustments for age, gender, clinical center, glucose tolerance status, body mass index, waist, triglycerides, and insulin resistance. Multiple linear regression analyses showed that the 4G/5G genotype explained very little of the variation in PAI-1 levels (0.63% in non Hispanic whites, 0.99% in Hispanics, and 2.37% in blacks), and interaction analyses revealed no significant differences in the relation of circulating PAI-1 levels to the 4G/5G genotype by ethnicity (P=0.4). CONCLUSIONS: We have shown ethnic differences in the PAI-1 4G/5G polymorphism along with corresponding differences in circulating PAI-1 levels. The association of the genotype with PAI 1 levels was seen consistently among all 3 ethnic groups and was unaffected by metabolic covariates, including insulin resistance. PMID- 12719279 TI - Effect of ezetimibe coadministered with atorvastatin in 628 patients with primary hypercholesterolemia: a prospective, randomized, double-blind trial. AB - BACKGROUND: Despite the established efficacy of statins, many patients do not achieve recommended LDL cholesterol (LDL-C) goals. Contributing factors may be inadequate dosing, increased risk for adverse effects with high-dose monotherapy, and increased potential for intolerance and adverse effects with combinations of available agents. METHODS AND RESULTS: In a double-blind study, 628 patients with baseline LDL-C 145 to 250 mg/dL and triglycerides < or =350 mg/dL were randomly assigned to receive 1 of the following for 12 weeks: ezetimibe (10 mg/d); atorvastatin (10, 20, 40, or 80 mg/d); ezetimibe (10 mg) plus atorvastatin (10, 20, 40, or 80 mg/d); or placebo. The primary efficacy end point was percentage reduction in LDL-C for pooled ezetimibe plus atorvastatin versus pooled atorvastatin treatment groups. Ezetimibe plus atorvastatin significantly improved LDL-C, HDL cholesterol (HDL-C), triglycerides, total cholesterol:HDL-C, and high sensitivity C-reactive protein (hs-CRP) compared with atorvastatin alone (P<0.01). Coadministration of ezetimibe provided a significant additional 12% LDL C reduction, 3% HDL-C increase, 8% triglyceride reduction, and 10% hs-CRP reduction versus atorvastatin alone. Ezetimibe plus atorvastatin provided LDL-C reductions of 50% to 60%, triglyceride reductions of 30% to 40%, and HDL-C increases of 5% to 9%, depending on atorvastatin dose. LDL-C reductions with ezetimibe plus 10 mg atorvastatin (50%) and 80 mg atorvastatin alone (51%) were similar. CONCLUSIONS: Ezetimibe plus atorvastatin was well tolerated, with a safety profile similar to atorvastatin alone and to placebo. When coadministered with atorvastatin, ezetimibe provided significant incremental reductions in LDL-C and triglycerides and increases in HDL-C. Coadministration of ezetimibe and atorvastatin offers a well-tolerated and highly efficacious new treatment option for patients with hypercholesterolemia. PMID- 12719280 TI - Accumulation of ultrasmall superparamagnetic particles of iron oxide in human atherosclerotic plaques can be detected by in vivo magnetic resonance imaging. AB - BACKGROUND: One of the features of high-risk atherosclerotic plaques is a preponderance of macrophages. Experimental studies with hyperlipidemic rabbits have shown that ultrasmall superparamagnetic particles of iron oxide (USPIOs) accumulate in plaques with a high macrophage content and that this induces magnetic resonance (MR) signal changes. The purpose of our study was to investigate whether USPIO-enhanced MRI can also be used for in vivo detection of macrophages in human plaques. METHODS AND RESULTS: MRI was performed on 11 symptomatic patients scheduled for carotid endarterectomy before and 24 (n=11) and 72 (n=5) hours after administration of USPIOs (Sinerem) at a dose of 2.6 mg Fe/kg. Histological and electron microscopical analyses of the plaques showed USPIOs primarily in macrophages within the plaques in 10 of 11 patients. Histological analysis showed USPIOs in 27 of 36 (75%) of the ruptured and rupture prone lesions and 1 of 14 (7%) of the stable lesions. Of the patients with USPIO uptake, signal changes in the post-USPIO MRI were observed by 2 observers in the vessel wall in 67 of 123 (54%) and 19 of 55 (35%) quadrants of the T2*-weighted MR images acquired after 24 and 72 hours, respectively. For those quadrants with changes, there was a significant signal decrease of 24% (95% CI, 33% to 15%) in regions of interest in the images acquired after 24 hours, whereas no significant signal change was found after 72 hours. CONCLUSIONS: Accumulation of USPIOs in macrophages in predominantly ruptured and rupture-prone human atherosclerotic lesions caused signal decreases in the in vivo MR images. PMID- 12719281 TI - Serum N-terminal pro-brain natriuretic peptide is a sensitive marker of myocardial dysfunction in AL amyloidosis. AB - BACKGROUND: Brain natriuretic peptide (BNP) is a marker of ventricular dysfunction and can be used to assess prognosis in heart failure and after myocardial infarction. Heart involvement is the most important prognostic factor and causes death in almost all patients with light-chain amyloidosis (AL). We investigated the prognostic value of NT-proBNP and its utility in monitoring amyloid heart dysfunction. METHODS AND RESULTS: NT-proBNP was quantified at diagnosis in 152 consecutive patients seen at the coordinating center of the Italian Amyloidosis Study Group (Pavia) from 1999 throughout 2001. Heart involvement was estimated on the basis of clinical signs, electrocardiography, and echocardiography. NT-proBNP concentrations differed in patients with (n=90, 59%) and without (n=62, 41%) heart involvement (median: 507.8 pmol/L versus 22.1 pmol/L, P=10(-7)). The best cutoff for heart involvement was at 152 pmol/L (sensitivity: 93.33%, specificity: 90.16%, accuracy: 92.05%) and distinguished two groups with different survival (P<0.001). The Cox multivariate model including NT-proBNP was better than models including echocardiographic and clinical signs of heart involvement. NT-proBNP appeared to be more sensitive than conventional echocardiographic parameters in detecting clinical improvement or worsening of amyloid cardiomyopathy during follow-up. CONCLUSIONS: NT-proBNP appeared to be the most sensitive index of myocardial dysfunction and the most powerful prognostic determinant in AL amyloidosis. It adds prognostic information for newly diagnosed patients and can be useful in designing therapeutic strategies and monitoring response. NT-proBNP is a sensitive marker of heart toxicity caused by amyloidogenic light chains. PMID- 12719282 TI - Human aortic valve calcification is associated with an osteoblast phenotype. AB - BACKGROUND: Calcific aortic stenosis is the third most common cardiovascular disease in the United States. We hypothesized that the mechanism for aortic valve calcification is similar to skeletal bone formation and that this process is mediated by an osteoblast-like phenotype. METHODS AND RESULTS: To test this hypothesis, we examined calcified human aortic valves replaced at surgery (n=22) and normal human valves (n=20) removed at time of cardiac transplantation. Contact microradiography and micro-computerized tomography were used to assess the 2-dimensional and 3-dimensional extent of mineralization. Mineralization borders were identified with von Kossa and Goldner's stains. Electron microscopy and energy-dispersive spectroscopy were performed for identification of bone ultrastructure and CaPO4 composition. To analyze for the osteoblast and bone markers, reverse transcriptase-polymerase chain reaction was performed on calcified versus normal human valves for osteopontin, bone sialoprotein, osteocalcin, alkaline phosphatase, and the osteoblast-specific transcription factor Cbfa1. Microradiography and micro-computerized tomography confirmed the presence of calcification in the valve. Special stains for hydroxyapatite and CaPO4 were positive in calcification margins. Electron microscopy identified mineralization, whereas energy-dispersive spectroscopy confirmed the presence of elemental CaPO4. Reverse transcriptase-polymerase chain reaction revealed increased mRNA levels of osteopontin, bone sialoprotein, osteocalcin, and Cbfa1 in the calcified valves. There was no change in alkaline phosphatase mRNA level but an increase in the protein expression in the diseased valves. CONCLUSIONS: These findings support the concept that aortic valve calcification is not a random degenerative process but an active regulated process associated with an osteoblast-like phenotype. PMID- 12719283 TI - Preliminary observations regarding angiographic pattern of restenosis after rapamycin-eluting stent implantation. AB - BACKGROUND: Restenosis after implantation of drug-eluting stents (DES) is a rare phenomenon, occurring more frequently peri-stent. METHODS AND RESULTS: We evaluated the pattern of restenosis occurring after implantation of DES in unselected lesions. From April 15 to December 6, 2002, we treated 368 patients with 735 lesions by using 841 rapamycin-eluting stents (Cypher, Cordis, a Johnson & Johnson Company). Mean baseline lesion length was 17.48+/-12.19 mm, and mean stent length was 27.59+/-14.02 mm. Follow-up ischemia-driven angiography was performed in 24 patients. Eleven patients had angiographic restenosis (> or =50% diameter stenosis) in 14 stented segments (stent and 5 mm proximal and distal to the stent). The pattern of restenosis in all 14 stented segments was focal, and in 6 of them it was multifocal, occurring inside the stents. Mean length of restenotic lesions was 5.62+/-1.90 mm, with a range from 2.54 to 8.44 mm. One multifocal restenosis involved also the distal stent margin. Intravascular ultrasound evaluation at follow-up, performed in 2 patients, showed significant lumen obstruction attributable to in-stent hyperplasia in both cases. Individual cases can be viewed in the Data Supplement. CONCLUSIONS: The pattern of restenotic lesions after rapamycin-eluting stent implantation was focal and mostly inside the stent. PMID- 12719284 TI - Receptor for AGE (RAGE) mediates neointimal formation in response to arterial injury. AB - BACKGROUND: Receptor for advanced-glycation end products (RAGE) and its ligands AGEs and S100/calgranulins have been implicated in a range of disorders. However, the role of RAGE/ligand interaction in neointimal hyperplasia after vascular injury remains unclear. METHODS AND RESULTS: We examined the expression of RAGE and its ligands after balloon injury of the carotid artery in both Zucker diabetic and nondiabetic rats. Using a soluble portion of the extracellular domain of RAGE, we determined the effects of suppressing RAGE/ligand interaction on vascular smooth muscle cell (VSMC) proliferation and neointimal formation after arterial injury. We demonstrate a significantly increased accumulation of AGE and immunoreactivities of RAGE and S100/calgranulins in response to balloon injury in diabetic compared with nondiabetic rats. Blockade of RAGE/ligand interaction significantly decreased S100-stimulated VSMC proliferation in vitro and bromodeoxyuridine (BrdU)-labeled proliferating VSMC in vivo, and suppressed neointimal formation and increased luminal area in both Zucker diabetic and nondiabetic rats. CONCLUSIONS: These findings indicate that RAGE/ligand interaction plays a key role in neointimal formation after vascular injury irrespective of diabetes status and suggest a novel target to minimize neointimal hyperplasia. PMID- 12719285 TI - Sirolimus (rapamycin) monotherapy prevents graft vascular disease in nonhuman primate recipients of orthotopic aortic allografts. AB - BACKGROUND: Delayed treatment with sirolimus (SRL) halts progression of graft vascular disease (GVD) in nonhuman primate (NHP) aortic allograft recipients. In this study, we investigated whether SRL monotherapy prevents the development of GVD. METHODS AND RESULTS: Pairs of 3-cm infrarenal aortic segments were exchanged between mixed lymphocyte reaction-mismatched, blood group-compatible NHPs (n=12). Six NHPs were untreated controls, and 6 were treated orally with SRL starting on the day of transplantation. Follow-up was 105 days. SRL doses were adjusted individually by assessing SRL blood concentrations, immune function, and clinical status. The severity of GVD was determined every 3 weeks by intravascular ultrasound, which quantified intimal area (IA) and intimal volume (IV) for the middle 1-cm graft segments. The mean+/-SEM SRL plasma levels were 14.5+/-9 ng/mL. In grafts from treated NHPs, IA and IV values on days 63, 84, and 105 were significantly lower than for controls (P<0.05 to P<0.001). On day 105, in the grafts from SRL-treated NHPs compared with grafts from controls, values (mean+/ SEM) were IA, 2.9+/-0.9 versus 5.5+/-0.7 mm2, P<0.001 and IV, 29.6+/-4.6 versus 55.2+/-2.8 mm3, P<0.001; IA and IV values for grafts from SRL-treated NHPs did not increase significantly between days 21 and 105. CONCLUSIONS: We show that SRL monotherapy prevented GVD in NHP aortic allograft recipients, suggesting the value of SRL for controlling GVD in clinical transplantation. PMID- 12719286 TI - Is there a vulnerable plaque? PMID- 12719287 TI - Vulnerable atherosclerotic plaque: a multifocal disease. PMID- 12719288 TI - The Emperor's clothes: in search of the vulnerable plaque. PMID- 12719289 TI - Images in cardiovascular medicine. Deep vein thrombosis demonstrated by magnetic resonance direct thrombus imaging. PMID- 12719290 TI - Cardiology patient page. Exercise testing and nuclear scanning. PMID- 12719291 TI - Images in cardiovascular medicine. Magnetic resonance assessment of cardiac function, infarct scar distribution, and ventricular remodeling in the setting of ischemic cardiomyopathy. PMID- 12719292 TI - Bicuspid aortic valve and coronary anomalies. PMID- 12719293 TI - Origin of smooth muscle progenitor cells: different conclusions from different models. PMID- 12719294 TI - Salt intake and sympathetic activity. PMID- 12719295 TI - Effect of rosiglitazone treatment on nontraditional markers of cardiovascular disease in patients with type 2 diabetes mellitus. PMID- 12719296 TI - Treatment of hypertrophic cardiomyopathy. PMID- 12719298 TI - The opportunity costs of war in Iraq. PMID- 12719301 TI - Increasing children's iron intake. PMID- 12719303 TI - Considering colorectal screening. PMID- 12719304 TI - Considering colorectal screening. PMID- 12719306 TI - Considering colorectal screening. PMID- 12719308 TI - Emergency docs or family physicians? PMID- 12719309 TI - Drug supply and drug abuse. PMID- 12719311 TI - Reference-based refinements. PMID- 12719312 TI - An open letter from Concerned Medical Students on Iraq. PMID- 12719314 TI - Characteristics of international medical graduates who applied to the CaRMS 2002 match. AB - BACKGROUND: International medical graduates are an important component of the Canadian physician workforce. For most international medical graduates, the principal route to obtaining a residency position in Canada is to apply through the second iteration of the Canadian Resident Matching Service (CaRMS) match. In order to help inform the work toward integrating unlicensed international medical graduates into Canada's health professional workforce, our objectives were to describe the demographic and educational characteristics of international medical graduate CaRMS applicants and identify their preferred clinical disciplines and practice locations. METHODS: A 37-item Web-based questionnaire survey was offered to all 659 international medical graduate second-iteration CaRMS 2002 applicants. We collected data on their demographic and educational background and preferred clinical discipline and practice location. Up to 2 follow-up email reminders were sent to nonrespondents. RESULTS: The survey response rate was 70.3% (463/659). Of the respondents, 71.9% had obtained their medical degree in Asia, the Middle East or Eastern Europe: 36.5% had graduated with a medical degree since 1994, and 17.3% since 1997. Most respondents (74.3%) were aged between 30 and 44 years. More than half (54.6%) had completed their medical education in English. Most (69.3%) had done postgraduate training outside Canada. Before coming to Canada, 42.8% had practised medicine for 1-5 years and 45.6% had practised for 6-20 years. The top 5 choices of clinical discipline in Canada were family medicine/general practice (45.6%), internal medicine (14.9%), surgery (7.3%), obstetrics/gynecology (6.7%) and pediatrics (4.8%). Of those who resided in the 4 Western provinces or Nova Scotia, between 76.8% and 86.7% preferred to stay in their own province, and 60%, 51.4% and 37% of those who resided in Newfoundland, Ontario or Quebec respectively preferred to practise in their own province. INTERPRETATION: Second-iteration international medical graduate CaRMS applicants are a heterogeneous group of physicians, some with substantial medical training and experience and others at an earlier stage of their medical career. PMID- 12719315 TI - Finding Canadian cancer clinical trials on the Internet: an exploratory evaluation of online resources. AB - BACKGROUND: Online information about the availability of clinical trials promises to enhance the accrual of patients into trials. The primary objective of our study was to assess the completeness of online databases of breast cancer clinical trials available in Canada. METHODS: Eligible online resources were identified through a search of MEDLINE (articles published from 1966 to January 2002), an Internet search with Google, examination of Web sites of cancer organizations and information from experts. Resources were included if they contained information about open, active cancer clinical trials available in Canada. Web sites reviewed were not limited to those based in Canada. For each eligible resource, the number of listed trials and the proportion of trials identified were calculated. RESULTS: Of 30 Web sites identified, 8 met all of the inclusion criteria; 3 were based in Canada and 5 in the United States. The total number of breast cancer trials identified as being available in Canada was 28. The Physician Data Query (PDQ) Clinical Trials Database of the US National Cancer Institute (cancer.gov/search/clinical_trials) identified 86% (24/28). The database of the National Cancer Institute of Canada Clinical Trials Group (NCIC CTG) (ctg.queensu.ca) identified 29% (8/28) of the available breast cancer trials. CenterWatch Clinical Trials Listing Service (centerwatch.com) identified 4% (1/28). INTERPRETATION: If the PDQ database included all of the NCIC CTG trials, it would become the most complete database of breast cancer clinical trials currently available in Canada. Online cancer data sources should strive to make access to clinical trials simpler and more reliable, particularly for residents of the country where the trial is to be conducted. PMID- 12719316 TI - Drugs in the news: an analysis of Canadian newspaper coverage of new prescription drugs. AB - BACKGROUND: Patients routinely cite the media, after physicians and pharmacists, as a key source of information on new drugs, but there has been little research on the quality of drug information presented. We assessed newspaper descriptions of drug benefits and harms, the nature of the effects described and the presence or absence of other important information that can add context and balance to a report about a new drug. METHODS: We looked at newspaper coverage in the year 2000 of 5 prescription drugs launched in Canada between 1996 and 2001 that received a high degree of media attention: atorvastatin, celecoxib, donepezil, oseltamivir and raloxifene. We searched 24 of Canada's largest daily newspapers for articles reporting at least one benefit or harm of any of these 5 drugs. We recorded the benefits and harms reported and analyzed how such information was presented; we also determined whether clinical or surrogate outcomes were mentioned; if and how drug effects were quantified; whether contraindications, other treatment options and costs were mentioned; and whether any information on affiliations of quoted interviewees and potential conflicts of interest was presented. RESULTS: Our search yielded 193 articles reporting at least one benefit or harm for 1 of the 5 drugs. All of the articles mentioned at least one benefit, but 68% (132/193) made no mention of possible side effects or harms. Only 24% (120/510) of mentions of drug benefits and harms presented quantitative information. In 26% (31/120) of cases in which drug benefits and harms were quantified, the magnitude was presented only in relative terms, which can be misleading. Overall, 62% (119/193) of the articles gave no quantification of the benefits or harms. Thirty-seven (19%) of the 193 articles reported only surrogate benefits. Other information needed for informed drug-related decisions was often lacking: only 7 (4%) of the articles mentioned contraindications, 61 (32%) mentioned drug costs, 89 (46%) mentioned drug alternatives, and 30 (16%) mentioned nondrug treatment options (such as exercise or diet). Sixty-two percent (120/193) of the articles quoted at least one interviewee. After exclusion of industry and government spokespeople, for only 3% (5/164) of interviewees was there any mention of potential financial conflicts of interest. Twenty-six percent (15/57) of the articles discussing a study included information on study funding. INTERPRETATION: Our results raise concerns about the completeness and quality of media reporting about new medications. PMID- 12719317 TI - Adaptation of Inuit children to a low-calcium diet. AB - For Inuit children, a traditional diet contains 20 mg of elemental calcium per day, well below the recommended daily intake. To identify alterations in intestinal or renal calcium absorption, 10 healthy Inuit children (5 to 17 years of age) were given a standardized calcium load (Pak test). Five had hypercalciuria (hyperabsorptive in 3 and renal leak in 2), a frequency markedly different from that for white children (p < 0.004) and not explained by calcitropic hormone and serum calcium levels, which were normal. There was a preponderance of the bb vitamin D receptor genotype (8 of 10 subjects; p < 0.01 for comparison with white populations). Dietary calcium absorption appeared to be more efficient in these Inuit children, with an increased frequency of hypercalciuria associated with the bb genotype. This may represent a genetic adaptation to dietary constraints and may predispose to nephrolithiasis or nephrocalcinosis if standard nutritional guidelines are followed. PMID- 12719318 TI - Methylprednisolone for acute spinal cord injury: not a standard of care. PMID- 12719319 TI - Alcohol: a recently identified risk factor for breast cancer. PMID- 12719320 TI - The challenge of an increasingly expensive blood system. PMID- 12719321 TI - Tumour necrosis factor-alpha inhibitors and the reactivation of latent tuberculosis infection. PMID- 12719322 TI - Galactic radiation exposure during commercial flights: is there a risk? PMID- 12719323 TI - Facial furuncle on 3-year-old boy camping in Ontario. PMID- 12719324 TI - Long-term, low-intensity warfarin after idiopathic venous thromboembolism. PMID- 12719327 TI - "Yuppie scans" unethical marketing ploy, radiologist says. PMID- 12719328 TI - Cardiac cases trigger advisory about smallpox vaccination. PMID- 12719329 TI - Kenyan sex workers key to AIDS vaccine? PMID- 12719332 TI - Alberta delivers new blow to prescription data mining. PMID- 12719333 TI - Korean to head World Health Organization. PMID- 12719335 TI - Medical students discover the reality of weapons of mass destruction. PMID- 12719336 TI - CMA presidency becomes a hot commodity. PMID- 12719337 TI - There's no end to debate about Montreal's new superhospitals. PMID- 12719339 TI - Much ballyhooed biohazard training yet to begin. PMID- 12719340 TI - "Hillbilly heroin" arrives in Cape Breton. PMID- 12719341 TI - Travelling $2.5-million exhibit to demystify DNA. PMID- 12719342 TI - BC seeks upper hand in national recruiting for rural MDs. PMID- 12719343 TI - Family matters. PMID- 12719344 TI - Dearth of dialysis feared in UK. PMID- 12719347 TI - Difficulties developing evidence-based approaches in learning disabilities. PMID- 12719348 TI - Vitamin and fatty acid supplements may reduce antisocial behaviour in incarcerated young adults. PMID- 12719349 TI - Once-daily atomoxetine may reduce attention deficit hyperactivity disorder symptoms in children and adolescents. PMID- 12719350 TI - Review: adderall may have a small advantage over methylphenidate in attention deficit hyperactivity disorder. PMID- 12719351 TI - 20mg transdermal selegiline daily may be effective and well tolerated in adults with major depression. PMID- 12719352 TI - Review: antidepressants and psychotherapy may be equally effective for promoting remission in major depressive disorder. PMID- 12719353 TI - Review: Low dose tricyclic antidepressants may be effective for adults with acute depressive disorder. PMID- 12719354 TI - Enhanced primary care may encourage remission in depression. PMID- 12719355 TI - Review: there is limited evidence about the effectiveness of interventions for treatment-refractory depression. PMID- 12719356 TI - 18-month maintenance treatment with sertraline may have sustained psychosocial benefits in chronic depression. PMID- 12719357 TI - Skills training plus exposure therapy may reduce post traumatic stress in women who experienced childhood abuse. PMID- 12719358 TI - Fluoxetine may prevent relapse in post traumatic stress disorder. PMID- 12719359 TI - Stress management programme may reduce hospital admissions among people with schizophrenia. PMID- 12719360 TI - Review: there is no strong evidence to support antipsychotic combination therapy in schizophrenia. PMID- 12719361 TI - Cognitive training may improve targeted cognitive functions in older adults. PMID- 12719362 TI - Review: cognitive behavioural interventions may be effective for chronic fatigue syndrome and chronic back pain. PMID- 12719363 TI - Group interpersonal psychotherapy may be as effective as group cognitive behavioural therapy for overweight people with binge eating disorder. PMID- 12719364 TI - Review: there is insufficient evidence for naltrexone maintenance treatment in opioid dependence. PMID- 12719365 TI - Skills training programmes for significant others may encourage unmotivated drug users to seek treatment. PMID- 12719366 TI - Almost half of people suffering traumatic brain injury may later be diagnosed with axis I disorders. PMID- 12719367 TI - Depressive symptoms in adolescence may increase the risk of psychiatric disorders in early adulthood. PMID- 12719368 TI - Individual and familial mental illness is associated with an increased risk of suicide in young people. PMID- 12719369 TI - There is little evidence that combined vaccination against measles, mumps, and rubella is associated with autism. PMID- 12719370 TI - Case-finding instruments may help identify depression in primary care. PMID- 12719371 TI - Many factors make family members caring for an elderly relative with dementia reluctant to use support services. PMID- 12719372 TI - Genetic dissection of anxiety in autoimmune disease. AB - Systemic lupus erythematosus (SLE), a complex multigenic disease, is characterized by hypergammaglobulinemia, autoantibody production and immune complex-type lupus nephritis. In addition to these signs and symptoms in SLE, there can be symptoms of neurological disorders, including anxiety. To clarify mechanisms governing the anxiety seen in lupus, we carried out genome-wide scans, and found that the region including interferon-alpha (IFN-alpha) on NZB chromosome 4 is significantly linked to the anxiety-like behavior seen in SLE prone New Zealand Black (NZB) x New Zealand White (NZW) F(1) (B/W F(1)) mice. This finding was confirmed by anxiety-like performances of mice with heterozygous NZB/NZW alleles in the susceptibility region onto the NZW background. In B/W F(1) mice, neuronal IFN-alpha levels were elevated, and blockade of the micro (1) opioid receptor or corticotropin-releasing hormone receptor 1, possible downstream effectors for IFN-alpha in the brain partially overcame the anxiety like behavior seen in the B/W F(1) mice. Consistently, neuronal corticotropin releasing hormone levels were higher in B/W F(1) than NZW mice. Furthermore, pretreatment of micro (1) opioid receptor antagonist abolished anxiety-like behaviour seen in IFN-alpha-treated NZW mice. Anxiety is shown to be mediated by multiple mediators. Our data suggest that a genetically determined endogenous excess amount of IFN-alpha in the brain may form one aspect of anxiety-like behavior seen in SLE-prone mice. PMID- 12719373 TI - Restoration of dystrophin expression in mdx muscle cells by chimeraplast-mediated exon skipping. AB - The most common types of dystrophin gene mutations that cause Duchenne muscular dystrophy (DMD) are large deletions that result in a shift of the translational reading frame. Such mutations generally lead to a complete absence of dystrophin protein in the muscle cells of affected individuals. Any therapeutic modality that could restore the reading frame would have the potential to substantially reduce the severity of the disease by allowing the production of an internally deleted, but partially functional, dystrophin protein as occurs in Becker muscular dystrophy (BMD). One approach to restoring the reading frame would be to alter the splicing of the pre-mRNA to produce an in-frame transcript. We have tested the ability of chimeric RNA/DNA oligonucleotides (chimeraplasts) to alter key bases in specific splice sequences in the dystrophin gene to induce exon skipping. Using cells from the mdx mouse as a model system, we show that chimeraplast-mediated base conversion in the intron 22/exon 23 splice junction induces alternative splicing and the production of in-frame transcripts. Interestingly, multiple alternative transcripts were induced by this targeted splice site mutation. Direct sequencing indicated that several of these were predicted to produce in-frame dystrophin transcripts with internal deletions. Indeed, multiple forms of dystrophin protein were observed by western blot analysis, and the functionality of the products was demonstrated by the restoration of expression and localization of a dystrophin-associated protein, alpha-dystroglycan, in differentiated cells. These data demonstrate that chimeraplasts can induce exon skipping by altering splice site sequences at the genomic level. As such, chimeraplast-mediated exon skipping has the potential to be used to transform a severe DMD phenotype into a much milder BMD phenotype. PMID- 12719374 TI - Association of a functional 17beta-estradiol sensitive IL6-174G/C promoter polymorphism with early-onset type 1 diabetes in females. AB - The type 1 diabetes mellitus (T1DM) candidate gene SNP IL6-174G/C was genotyped in 253 Danish T1DM families (1129 individuals). TDT analysis demonstrated linkage in the presence of association between the IL6-174C allele and T1DM in the 416 T1DM offspring, P(tdt)=0.04. Gender conditioned TDT analyses revealed that linkage and association with T1DM were present in females exclusively; P(tdt)=6.5 x 10(-4) and P(tdt)=2.4 x 10(-4), respectively. Random transmission of the IL6 174C/G alleles was found in T1DM males, non-T1DM males and non-T1DM females; all P(tdt)>/=0.37. Heterogeneity analyses (T1DM versus non-T1DM females) excluded preferential meiotic segregation in females, P=4.6 x 10(-3), and demonstrated differences in the transmission patterns between female and male T1DM offspring, P=5.1 x 10(-3). The IL6-174 CC genotype was associated with younger age at onset of T1DM in females (P=0.002). The impact of 17beta-estradiol (E(2)) on the IL6 174G/C variants was investigated by reporter studies. The PMA stimulated activity of the T1DM risk IL6-174C variant exceeded that of the T1DM protective IL6-174G variant by approximately 70% in the absence of E(2) (P(c)=0.004), but not with E(2) present (P(c)=0.12). The PMA stimulated activity of the IL6-174G variant was repressed without E(2) present, but was derepressed by addition of E(2), P(c)=0.024. In contrast, the PMA stimulated IL6-174C activity was unaffected by E(2) as were the constitutive activities of the IL6-174G/C variants. In conclusion, higher IL6 promoter activity may confer risk to T1DM in very young females. This excess risk is negated with increasing age, possibly by the increasing E(2) levels in puberty. PMID- 12719375 TI - New type of disease causing mutations: the example of the composite exonic regulatory elements of splicing in CFTR exon 12. AB - The increase in genome scanning data, derived from clinical genetics practice, is producing a wealth of information on human sequence variability. The critical issue is to identify if a given nucleotide change results in a benign polymorphism or a disease-causing mutation. We have focused on one specific gene expression step, pre-mRNA processing, where we can functionally define the effect of nucleotide changes and in turn the patient's mutation can shed light on the basic pre mRNA splicing mechanisms. Our results show that several nucleotide changes in CFTR exon 12 induce a variable extent of exon skipping that leads to reduced levels of normal transcripts. This is the case in both natural mutations D565G and G576A (the latter having previously considered a neutral polymorphism) and several site-directed silent substitutions. We demonstrate here that this phenomenon is due to the interference with a new regulatory element that we have named composite exonic regulatory element of splicing (CERES). The effect of single nucleotide substitutions at CERES cannot be predicted by neither SR matrices nor enhancer identification. The recognition and characterization of splicing abnormalities, caused by exon sequence variations at CERES elements, may represent a frequent disease-causing mechanism that also relates to the phenotypic variability. Our results indicate that even the most benign looking polymorphism in an exon cannot be ignored as it may affect the splicing process. Hence, appropriate functional splicing assays should be included in genotype screenings to distinguish between polymorphisms and pathogenic mutations. PMID- 12719376 TI - Functional polymorphisms in the paternally expressed XLalphas and its cofactor ALEX decrease their mutual interaction and enhance receptor-mediated cAMP formation. AB - The paternally expressed extra-large stimulatory G protein gene (XLalphas) is a splice variant of the stimulatory G-protein gene (Gsalpha) consisting of XL-exon1 and exons 2-13 of Gsalpha. A second open reading frame (ORF) in XL-exon1, that completely overlaps the XL-domain ORF, encodes ALEX, which is translated from the XLalphas mRNA and binds the XL-domain of XLalphas. We previously demonstrated that a paternally inherited functional polymorphism in XL-exon1, consisting of a 36 bp insertion and two nucleotide substitutions, is associated with Gs hyperfunction in platelets, leading to an increased trauma-related bleeding tendency and is accompanied by neurological problems and brachydactyly in two families. Here, we describe eight additional patients with brachydactyly, who inherited the same XLalphas polymorphism paternally and who show Gs hyperfunction in their platelets and fibroblasts. All carriers also have an elongated ALEX protein, as a consequence of the paternally inherited insertion. The in vitro interaction between the two elongated XLalphas and ALEX proteins is markedly reduced. Moreover, XLalphas or ALEX can be co-immunoprecipitated with an antibody against either ALEX or XLalphas in platelets from a control but hardly from patients with the XLalphas/ALEX insertion. In contrast to the strong interaction between the two wild-type proteins, we suggest that this defective association results in unimpeded receptor-stimulated activation of XLalphas. The paternally inherited double XLalphas/ALEX functional polymorphism is also associated with elevated platelet membrane Gsalpha protein levels. Both phenomena contribute to increased Gs signaling in patients with platelet hypersensitivity towards Gs agonists and may be accompanied by neurological problems or growth deficiency. PMID- 12719377 TI - Identification of a novel lipase gene mutated in lpd mice with hypertriglyceridemia and associated with dyslipidemia in humans. AB - Triglyceride (TG) metabolism is crucial for whole body and local energy homeostasis and accumulating evidence suggests an independent association between plasma TG concentration and increased atherosclerosis risk. We previously generated a mouse insertional mutation lpd (lipid defect) whose phenotype included elevated plasma TG and hepatic steatosis. Using shotgun sequencing (approximately 500 kb) and bioinformatics, we have now identified a novel lipase gene lpdl (lpd lipase) within the lpd locus, and demonstrate the genetic disruption of exon 10 of lpdl in the lpd mutant locus. lpdl is highly expressed in the testis and weakly expressed in the liver of 2-week old mice. Human LPDL cDNA was subsequently cloned, and was found to encode a 460AA protein with 71% protein sequence identity to mouse lpdl and approximately 35% identity to other known lipases. We next sequenced the human LPDL gene exons in hypertriglyceridemic subjects and normal controls, and identified seven SNPs within the gene exons and six SNPs in the adjacent introns. Two hypertriglyceridemic subjects were heterozygous for a rare DNA variant, namely 164G>A (C55Y), which was absent from 600 normal chromosomes. Two other coding SNPs were associated with variation in plasma HDL cholesterol in independent normolipidemic populations. Using bioinformatics, we identified another novel lipase designated LPDLR (for 'LPDL related lipase'), which had 44% protein sequence identity with LPDL. Together with the phospholipase gene PSPLA1, LPDL and LPDLR form a new lipase gene subfamily, which is characterized by shortened lid motif. Study of this lipase subfamily may identify novel molecular mechanisms for plasma and/or tissue TG metabolism. PMID- 12719378 TI - A very long-chain acyl-CoA synthetase-deficient mouse and its relevance to X linked adrenoleukodystrophy. AB - X-linked adrenoleukodystrophy (X-ALD) is a neurodegenerative and endocrine disorder resulting from mutations in ABCD1 which encodes a peroxisomal membrane protein in the ATP binding cassette superfamily. The biochemical signature of X ALD is increased levels of saturated very long-chain fatty acids (VLCFA; carbon chains of 22 or more) in tissues and plasma that has been associated with decreased peroxisomal very long-chain acyl-CoA synthetase (VLCS) activity and decreased peroxisomal VLCFA beta-oxidation. It has been hypothesized that ABCD1, which has no demonstrable VLCS activity itself, has an indirect effect on peroxisomal VLCS activity and VLCFA beta-oxidation by transporting fatty acid substrates, VLCS protein or some required co-factor into peroxisomes. Here we report the characterization of a Vlcs knockout mouse that exhibits decreased peroxisomal VLCS activity and VLCFA beta-oxidation but does not accumulate VLCFA. The XALD/Vlcs double knockout mouse has the biochemical abnormalities observed in the individual knockout mice but does not display a more severe X-ALD phenotype. These data lead us to conclude that (1) VLCFA levels are independent of peroxisomal fatty acid beta-oxidation, (2) there is no ABCD1/VLCS interaction and (3) the common severe forms of X-ALD cannot be modeled by decreasing peroxisomal VLCS activity in the XALD mouse. PMID- 12719379 TI - Prestin, a cochlear motor protein, is defective in non-syndromic hearing loss. AB - Prestin, a membrane protein that is highly and almost exclusively expressed in the outer hair cells (OHCs) of the cochlea, is a motor protein which senses membrane potential and drives rapid length changes in OHCs. Surprisingly, prestin is a member of a gene family, solute carrier (SLC) family 26, that encodes anion transporters and related proteins. Of nine known human genes in this family, three (SLC26A2, SLC26A3 and SLC26A4) are associated with different human hereditary diseases. The restricted expression of prestin in OHCs, and its proposed function as a mechanical amplifier, make it a strong candidate gene for human deafness. Here we report the cloning and characterization of four splicing isoforms for the human prestin gene (SLC26A5a, b, c and d). SLC26A5a is the predominant form of prestin whereas the others showed limited distribution associated with certain developmental stages. Based on the functional importance of prestin we screened for possible mutations involving the prestin gene in a group of deaf probands. We have identified a 5'-UTR splice acceptor mutation (IVS2-2A>G) in exon 3 of the prestin gene, which is responsible for recessive non syndromic deafness in two unrelated families. In addition, a high frequency of heterozygosity for the same mutation was observed in these subjects, suggesting the possibility of semi-dominant influence of the mutation in causing hearing loss. Finally, the observation of this mutation only in the Caucasian probands indicated an association with a specific ethnic background. This study thereby reveals an essential function of prestin in human auditory processing. PMID- 12719380 TI - Congenital hydrocephalus in hy3 mice is caused by a frameshift mutation in Hydin, a large novel gene. AB - The autosomal-recessive mutation hydrocephalus3 (hy3) results in lethal communicating hydrocephalus with perinatal onset. We recently described a hydrocephalus-inducing transgenic insertional mutation, OVE459, which represents a new allele of hy3. Direct cDNA selection performed on a wild-type mouse BAC clone spanning the OVE459 insertion locus on chromosome 8 led to the identification of two novel candidate genes, Hydin and Vac14. The transgene insertion resulted in a rearrangement of Hydin exons in OVE459 mice. Hydin consists of at least 86 exons spanning over 340 kb of genomic DNA. The full length Hydin transcript is nearly 16 kb, encoding a putative 5099 amino acid protein. Northern analysis revealed a marked reduction of Hydin mRNA in both OVE459 and hy3 homozygotes relative to wild-type littermates. A single CG base pair deletion in exon 15 of Hydin was discovered specifically in mice carrying the spontaneous hy3 mutant allele. This deletion creates a premature termination signal two codons downstream of the mutation, likely resulting in the loss of 89% of the full-length gene product. Within the neonatal brain, Hydin expression is confined to the ciliated ependymal cell layer lining the lateral, third and fourth ventricles. Other sites of Hydin expression include the ciliated epithelial cells lining the bronchi and oviduct, as well as in the developing spermatocytes in the testis. The Hydin gene product is not closely related to any previously identified protein, with the exception of a 314 amino acid domain with homology to caldesmon, an actin-binding protein, suggesting an interaction with the cytoskeleton. PMID- 12719381 TI - A homozygous splicing mutation causing a depletion of skeletal muscle RYR1 is associated with multi-minicore disease congenital myopathy with ophthalmoplegia. AB - The ryanodine receptor (RYR1) is an essential component of the calcium homeostasis of the skeletal muscle in mammals. Inactivation of the RYR1 gene in mice is lethal at birth. In humans only missense and in-frame mutations in the RYR1 gene have been associated so far with various muscle disorders including malignant hyperthermia, central core disease and the moderate form of multi minicore disease (MmD). We identified a cryptic splicing mutation in the RYR1 gene that resulted in a 90% decrease of the normal RYR1 transcript in skeletal muscle. The 14646+2.99 kb A-->G mutation was associated with the classical form of MmD with ophthalmoplegia, whose genetic basis was previously unknown. The mutation present at a homozygous level was responsible for a massive depletion of the RYR1 protein in skeletal muscle. The mutation was not expressed in lymphoblastoid cells, pointing toward a tissue specific splicing mechanism. This first report of an out-of-frame mutation that affects the amount of RYR1 raised the question of the amount of RYR1 needed for skeletal muscle function in humans. PMID- 12719382 TI - FOXC2 haploinsufficient mice are a model for human autosomal dominant lymphedema distichiasis syndrome. AB - Lymphedema-distichiasis (LD) (OMIM 153400) is a rare autosomal-dominant condition characterized by pubertal onset of lower limb lymphedema and an aberrant second row of eyelashes arising from the meibomian glands. In some patients cardiac, skeletal and other defects coexist. We previously identified inactivating, nonsense and frameshift mutations in the forkhead transcription factor FOXC2 in affected members of LD families. To further delineate the relationship of FOXC2 deficiency to the clinical (and lymphangiodysplastic) phenotype in this syndrome, we performed dynamic lymphatic imaging and immunohistochemical examination of lymphatic tissues in mice heterozygous (+/-) for a targeted disruption of Foxc2. Adult heterozygote mice characteristically exhibited a generalized lymphatic vessel and lymph node hyper plasia and rarely exhibited hindlimb swelling. Retrograde lymph flow through apparently incompetent interlymphangion valves into the mesenteric nodes, intestinal wall and liver was also observed. In addition, Foxc2 +/- mice uniformly displayed distichiasis. We conclude that Foxc2 haploinsufficient mice mimic closely the distinctive lymphatic and ocular phenotype of LD patients. Furthermore, the craniofacial, cardiovascular and skeletal abnormalities sometimes associated with LD have previously been shown to be fully penetrant in homozygous Foxc2 null mice. This Foxc2 mutant mouse thus provides an ideal model for exploring molecular mechanisms and physiologic events in mesenchymal differentiation associated with lymphatic growth and development and the clinical abnormalities seen in human LD syndrome. PMID- 12719383 TI - Molecular mechanisms underlying limb anomalies associated with cholesterol deficiency during gestation: implications of Hedgehog signaling. AB - Human disorders caused by inborn errors of cholesterol biosynthesis are characterized by dysmorphogenesis of multiple organs. This includes limb malformations that are observed at high frequency in some disorders, such as the Smith-Lemli-Opitz syndrome, indicating a pivotal role of cholesterol in limb morphogenesis. Recently, it has been demonstrated that cholesterol can modulate the activity of the Hedgehog proteins, that act as morphogens to regulate the precise patterning of many embryonic structures, among which the developing limbs. To provide insight in the functions of cholesterol during limb development and in the potential role of Hedgehog signaling in the genesis of limb defects, we developed an in vivo rat model of cholesterol deficiency. We show here that treatment with Triparanol, a distal inhibitor of cholesterol biosynthesis, induced patterning defects of the autopod at high frequency, including pre-axial syndactyly and post-axial polydactyly, thus reproducing limb anomalies frequently observed in humans. Using in situ hybridization, we show that these malformations originate from a modification of Sonic Hedgehog signaling in the limb bud at 13 days post-coitum, leading to a deficiency of the anterior part of the limb. This deficiency results in an imbalance of Indian Hedgehog expression in the forming cartilage, ultimately leading to reduced interdigital apoptosis and syndactyly. Our study thus unravels the molecular mechanisms underlying the genesis of limb defects associated with cholesterol deficiency in rodents, and most probably in humans. PMID- 12719384 TI - Genome-wide linkage analysis of bronchodilator responsiveness and post bronchodilator spirometric phenotypes in chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is a common, complex disease associated with significant and increasing morbidity and mortality. The cardinal feature of COPD is persistent airflow obstruction, measured by reductions in quantitative spirometric indices including forced expiratory volume at one second (FEV(1)) and the ratio of FEV(1) to forced vital capacity (FEV(1)/FVC). However, many patients have substantial improvement in spirometric measures with inhaled bronchodilator medications, and bronchodilator responsiveness (BDR) has been associated with disease severity and progression. To identify susceptibility loci for BDR phenotypes, we performed a 9 cM genome scan in 72 pedigrees (n=560 members) ascertained through probands with severe, early-onset COPD. Multipoint variance component linkage analysis was performed for quantitative phenotypes including BDR measures and post-bronchodilator FEV(1) and FEV(1)/FVC. Post bronchodilator FEV(1) was linked to multiple regions, most significantly to markers on chromosome 8p (LOD=3.30) and 1q (LOD=2.24). Post-bronchodilator FEV(1)/FVC was also linked to multiple regions, most significantly to markers on chromosome 2q (LOD=4.42) and 1q (LOD=2.52). When compared with pre-bronchodilator spirometric indices, the post-bronchodilator values demonstrated increased evidence of linkage in multiple genomic regions. In particular, the LOD score for the 8p linkage to FEV(1) roughly doubled from 1.58 to 3.30. Candidate regions on chromosomes 4p (LOD=1.28), 4q (LOD=1.56), and 3q (LOD=1.50) gave the strongest evidence for linkage to BDR measures. Our results provide evidence for significant linkage to airflow obstruction susceptibility loci on chromosomes 2q and 8p, and further suggest that post-bronchodilator spirometric measures are optimal phenotypes for COPD genetic studies. This study has also identified several genomic regions that could contain loci regulating BDR in early-onset COPD families. PMID- 12719385 TI - Obtaining consent for neonatal research. PMID- 12719386 TI - Nasal CPAP for neonates: what do we know in 2003? AB - Despite the acknowledged clinical usefulness of nasal CPAP, uncertainties regarding aspects of its application remain. Clinical indications for the application of nasal CPAP vary greatly between institutions. Furthermore, defining the optimal nasal CPAP system is complicated by the multiplicity of nasal CPAP devices and techniques available to the clinician. This review aims to identify what we know about nasal CPAP and what important questions remain. PMID- 12719388 TI - Neonatal meningitis. AB - Twelve years ago an annotation was published in Archives of Disease in Childhood regarding the antibiotic treatment of suspected neonatal meningitis. The authors recommended the use of cephalosporins rather than chloramphenicol and advocated intraventricular aminoglycoside treatment in selected cases. They noted the absence of clinical trials with third generation cephalosporins that showed an improvement in mortality or neurological outcome. PMID- 12719389 TI - Long term outcome of neonatal meningitis. AB - OBJECTIVES: To quantify long term impairment after neonatal meningitis. DESIGN: Longitudinal case-control study over 9-10 years. SUBJECTS AND METHODS: A total of 111 children who had suffered neonatal meningitis were seen and compared with 113 matched controls from their birth hospital and 49 controls from general practices. Assessments included the WISC III(UK), movement assessment battery for children (mABC), audiometry, vision testing, and social and medical data. Statistical analysis was by multiple regression, analysis of variance, and chi(2) tests. RESULTS: Some 10.8% of cases had a severe and 9% a moderate overall outcome compared with 0% and 1.8% for the hospital controls. The mean intelligence quotient (IQ) of the cases (88.8) was significantly less than that of the hospital controls (99.4) or the GP controls (99.6). The mABC score was significantly worse for the cases (7.08) than the hospital (5) or GP (4) controls. Some 3.6% of cases had sensorineural hearing loss, 2.7% had persisting hydrocephalus; no controls did. Some 5.4% of cases and 1.7% of hospital controls had treatment for seizures. CONCLUSIONS: Severe neurodisability and milder motor and psychometric impairment result from neonatal meningitis. Both clinical follow up and comprehensive developmental assessment are needed after this disease. PMID- 12719390 TI - Congenital brachial palsy: incidence, causes, and outcome in the United Kingdom and Republic of Ireland. AB - OBJECTIVES: To determine the incidence and study the causes and outcome of congenital brachial palsy (CBP). DESIGN: Active surveillance of newborn infants using the British Paediatric Surveillance Unit notification system and follow up study of outcome at 6 months of age. SETTING: The United Kingdom and Republic of Ireland. PARTICIPANTS: Newborn infants presenting with a flaccid paresis of the arm (usually one, rarely both) born between April 1998 and March 1999. MAIN OUTCOME MEASURES: Extent of the lesion at birth and degree of recovery at 6 months of age. FINDINGS: There were 323 confirmed cases giving an incidence of 0.42 per 1000 live births (1 in 2300). Significant associated risk factors in comparison with the normal population were shoulder dystocia (60% v 0.3%), high birth weight with 53% infants weighing more than the 90th centile, and assisted delivery (relative risk (RR) 3.4, 95% confidence interval (CI) 2.9 to 3.9, p = 0.0001). There was a considerably lower risk of CBP in infants delivered by caesarean section (RR 7, 95% CI 2 to 56, p = 0.002). At about 6 months of age, about half of the infants had recovered fully, but the remainder showed incomplete recovery including 2% with no recovery. The relative risk of partial or no recovery in infants with extensive lesions soon after birth compared with those with less extensive lesions was 11.28 (95% CI 2.38 to 63.66, p = 0.000005). CONCLUSIONS: The incidence of CBP in the United Kingdom and Republic of Ireland is strikingly similar to that previously reported nearly 40 years ago. Most cases are due to trauma at delivery, which is not necessarily excessive or inappropriate. Given the uncertainty about the appropriate management of these infants, serious consideration should be given to a formal clinical trial of microsurgical nerve repair. PMID- 12719391 TI - Health and school performance of teenagers born before 29 weeks gestation. AB - OBJECTIVE: To ascertain the health and school performance of teenagers born before 29 weeks gestation (extremely low gestational age (ELGA)) and to compare those in mainstream school with classroom controls. METHODS: Three geographically defined cohorts of babies born in 1983 and 1984 were traced at the age of 15-16 years. Their health, abilities, and educational performance were ascertained using postal questionnaires to the teenagers themselves, their parents, their general practitioners, and the teachers of those in mainstream school. Identical questionnaires were sent to classroom controls. RESULTS: Of the 218 teenagers surviving to the age of 16 years, information was obtained on 179. Of these, 29 were in special schools and 150 in mainstream school, 10 of whom had severe motor or sensory impairment. Using the Child Health Questionnaire, parents of teenagers in mainstream school reported a higher incidence of problems than controls in physical functioning (difference in mean scores 9.0 (95% confidence interval (CI) 4.9 to 13.1)) and family life (difference in mean scores for family cohesion 7.0 (95% CI 1.6 to 12.4)). In all areas of learning, teachers rated the ability of the ELGA teenagers in mainstream school lower than the control group. Parents of teenagers in special schools reported a higher rate of problems in most areas. CONCLUSIONS: One in six ELGA survivors at age 16 years have severe disabilities and are in special schools. Most ELGA survivors are in mainstream school and are coping well as they enter adult life, although some will continue to need additional health, educational, and social services. PMID- 12719392 TI - Non-viable delivery at 20-23 weeks gestation: observations and signs of life after birth. AB - OBJECTIVE: To describe the outcome of labour, signs of life at birth, and duration of survival after delivery at 20-23 weeks gestation. DESIGN: An observational study using data from the Confidential Enquiry into Stillbirths and Deaths in Infancy 1995-2000. SETTING: All deliveries to mothers resident in Trent Health Region. PATIENTS: 1306 babies delivered at 20-23 weeks gestation. RESULTS: Termination of pregnancy accounted for 33% of deliveries at 20-23 weeks; these were excluded from further analysis. Spontaneous delivery occurred at a frequency of 2.5/1000 deliveries; 30% died before the onset of labour, 27% died during labour, and 35% showed signs of life at birth. Of the latter, 8% were not registered as statutory live births. Of the live born infants, the largest group (39%) had a heart beat but no other signs of life. There was no trend for infants of lower gestation to show fewer signs of life. Duration of survival varied widely (median 60 minutes at 20-22 weeks), and this did not increase with gestation until 23 weeks (median six hours), probably because of selective treatment. Survival curves are presented for each gestation group. At 23 weeks, 4.5% survived to 1 year of age; all were > 500 g birth weight. Below 23 weeks gestation, none survived, and 94% had died within 4 hours of age. CONCLUSIONS: This information on surviving labour, signs of life at birth, duration of survival, and birth weight at 20-23 weeks gestation should help decision making in the management of pre-viable delivery. PMID- 12719393 TI - Low field strength magnetic resonance imaging of the neonatal brain. AB - BACKGROUND: Magnetic resonance (MR) imaging of the neonate has been restricted by the need to transport the sick baby to the large magnetic resonance scanners and often the need for sedation or anaesthesia in order to obtain good quality images. Ultrasound is the reference standard for neonatal imaging. OBJECTIVE: To establish a dedicated neonatal MR system and compare the clinical usefulness of MR imaging with ultrasound imaging. DESIGN: Prospective double blind trial. SETTING: Neonatal intensive care unit, Sheffield. MAIN OUTCOME MEASURES: Imaging reports. PATIENTS: 134 premature and term babies. RESULTS: In 56% of infants with pathology suspected on clinical grounds, MR provided additional useful clinical information over and above that obtained with ultrasound. CONCLUSION: Infants can be safely imaged by dedicated low field magnetic resonance on the neonatal intensive care unit without the need for sedation at a cost equivalent to ultrasound. PMID- 12719394 TI - Proinflammatory and anti-inflammatory cytokine responses in preterm infants with systemic infections. AB - OBJECTIVE: A prospective study to investigate the pattern of proinflammatory and anti-inflammatory cytokine responses in preterm infants with systemic infection. METHODS: Very low birthweight infants in whom infection was suspected when they were > 72 hours of age were eligible. A full sepsis screen was performed in each episode. Key cytokines of both proinflammatory and anti-inflammatory pathways, including interleukin (IL) 2, IL4, IL5, IL6, IL10, interferon (IFN) gamma, and tumour necrosis factor (TNF) alpha, were measured at 0 (at the time of sepsis evaluation), 24, and 48 hours by flow cytometric analysis or immunoassay. RESULTS: Thirty seven of the 127 episodes of suspected clinical sepsis were proven infection or necrotising enterocolitis. Both proinflammatory (IL2, IL6, IFNgamma, TNFalpha) and anti-inflammatory (IL4, IL10) cytokines were significantly increased in infected infants compared with non-infected infants. Significant correlations were observed between IL6 and TNFalpha or IL10 as well as IL10 and IFNgamma in infected infants. In the subgroup analysis, plasma IL6, IL10, and TNFalpha concentrations, and IL10/TNFalpha and IL6/IL10 ratios were significantly elevated in patients with disseminated intravascular coagulation compared with infected infants without. The IL10/TNFalpha ratios had decreased significantly 48 hours after the onset, whereas the IL6/IL10 ratio showed only a non-significant decreasing trend. Further, the IL6/IL10 ratio in the deceased infant was disproportionally increased at presentation and continued to increase despite treatment. CONCLUSION: The results indicate that the counter-regulatory mechanism between the proinflammatory and anti-inflammatory cytokine pathways is probably operational in preterm infants of early gestation. High plasma IL6, IL10, and TNFalpha concentrations, and IL10/TNFalpha and IL6/IL10 ratios signify severe infection, but transiently elevated plasma IL10 concentration or IL10/TNFalpha ratio does not necessarily indicate a poor prognosis. PMID- 12719395 TI - Insulin in human milk: postpartum changes and effect of gestational age. AB - OBJECTIVE: To determine if human milk insulin (HMI) concentrations are affected by gestational age and postnatal age. DESIGN AND SETTING: An observational study carried out in a level III neonatal intensive care unit. Insulin concentrations were determined in human milk of 90 parturient mothers who delivered between 30 and 41 weeks gestation. Samples were collected on days 3 and 10 after delivery. RESULTS: HMI concentrations for mothers of preterm infants were not significantly different from those of full term infants, on either day 3 or 10 post partum. When results for all 90 mothers were pooled, regardless of gestational age, HMI concentration fell significantly from day 3 to day 10 (50.1 (34.6) v 41.1 (28.5) microU/ml; p = 0.01; mean (SD)). However, this decrease was only significant for mothers delivering at term (37-41 weeks). CONCLUSIONS: HMI concentrations were not influenced by gestational age at delivery. They decreased post partum, mainly in mothers of term infants. The postnatal changes in HMI concentrations and the effects of oral insulin on the immature intestinal mucosa warrant further investigation. PMID- 12719397 TI - Cardiac output, pulmonary artery pressure, and patent ductus arteriosus during therapeutic cooling after global hypoxia-ischaemia. AB - OBJECTIVE: To assess by Doppler echocardiography the effects of 24 hours of whole body mild hypothermia compared with normothermia on cardiac output (CO), pulmonary artery pressure (PAP), and the presence of a persistent ductus arteriosus (PDA) after a global hypoxic-ischaemic insult in unsedated newborn animals. DESIGN: Thirty five pigs (mean (SD) age 26.6 (12.1) hours and weight 1.6 (0.3) kg) were anaesthetised with halothane, mechanically ventilated, and subjected to a 45 minute global hypoxic-ischaemic insult. At the end of hypoxia, halothane was stopped; the pigs were randomised to either normathermia (39 degrees C) or hypothermia (35 degrees C) for 24 hours. Rewarming was carried out for 24-30 hours followed by 42 hours of normothermia. Unanaesthetised pigs were examined with a VingMed CFM 750 ultrasound scanner before and 3, 24, 30, and 48 hours after the hypoxic-ischaemic insult. Aortic valve diameter, forward peak flow velocities across the four valves, and the occurrence of a PDA were measured. Tricuspid regurgitation (TR) velocity was used to estimate the PAP. Stroke volume was calculated from the aortic flow. RESULTS: Twelve animals (seven normothermic, five hypothermic) had a PDA on one or more examinations, which showed no association with cooling or severity of insult. There were no differences in stroke volume or TR velocity between the hypothermic and normothermic animals at any time point after the insult. CO was, however, 45% lower at the end of cooling in the subgroup of hypothermic pigs that had received a severe insult compared with the pigs with mild and moderate insults. CO and TR velocity were transiently increased three hours after the insult: 0.38 (0.08) v 0.42 (0.08) litres/min/kg (p = 0.007) for CO; 3.0 (0.42) v 3.4 (0.43) m/s (p < 0.0001) for TR velocity (values are mean (SD)). CONCLUSIONS: The introduction of mild hypothermia while the pigs were unsedated did not affect the incidence of PDA nor did it lead to any changes in MABP or PAP. Stroke volume was also unaffected by temperature, but hypothermic piglets subjected to a severe hypoxic ischaemic insult had reduced CO because the heart rate was lower. Global hypoxia ischaemia leads to similar transient increases in CO and estimated PAP in unsedated normothermic and hypothermic pigs. There were no signs of metabolic compromise in any subgroup, suggesting that 24 hours of mild hypothermia had no adverse cardiovascular effect. PMID- 12719396 TI - Respiratory responses to hypoxia/hypercapnia in small for gestational age infants influenced by maternal smoking. AB - AIM: To determine any variation in the respiratory responses to hypoxia/hypercapnia of infants born small for gestational age (SGA) to smoking and to non-smoking mothers. METHODS: A total of 70 average for gestational age (AGA) infants (>36 weeks gestation, >2500 g, >25th centile for gestational age, and no maternal smoking), and 47 SGA infants (<10th centile for gestational age) were studied at 1 and 3 months of age, in quiet and active sleep. Respiratory test gases were delivered through a Perspex hood to simulate face down rebreathing by slowly allowing the inspired air to be altered to a CO(2) maximum of 5% and O(2) minimum of 13.5%. The change in ventilation with inspired CO(2) was measured over 5-6 minutes of the test. The slope of a linear curve fit relating inspired CO(2) to the logarithm of ventilation was taken as a quantitative measure of ventilatory asphyxial sensitivity (VAS). RESULTS: There was no significant difference in VAS between the AGA and SGA infants (0.25 v 0.24). However within the SGA group, VAS was significantly higher (p = 0.048) in the infants whose mothers smoked during pregnancy (0.26 (0.01); n = 24) than in those that did not (0.23 (0.01); n = 23). The change in minute ventilation was significantly higher in the smokers than the non-smokers group (141% v 119%; p = 0.03) as the result of a significantly larger change in respiratory rate (8 v 4 breaths/min; p = 0.047) but not tidal volume. CONCLUSIONS: Maternal smoking appears to be the key factor in enhancing infants' respiratory responses to hypoxia/hypercapnia, irrespective of gestational age. PMID- 12719398 TI - Updated gestational age specific birth weight, crown-heel length, and head circumference of Chinese newborns. AB - OBJECTIVE: To construct gestation specific standards of birth weight, crown-heel length, and head circumference of Chinese infants. DESIGN: A prospective cross sectional population study. METHODS: The birth weight, crown-heel length, and head circumference were prospectively measured using standard equipment in newborns delivered at 24-42 weeks gestation in the maternity units of 10 public hospitals and two private hospitals in Hong Kong. The findings were used to construct gestation specific standards of these variables. The LMS method using maximum penalised likelihood was used to perform model fitting. The results were compared with those obtained from a cohort of infants born in the same locality between 1982 and 1986. RESULTS: From October 1998 to September 2000, a total of 10 032 infants were measured, representing 9.6% of the total deliveries in Hong Kong during that period. An extra 307 infants with gestation < or = 35 weeks were recruited from October 2000 to June 2001. Each of the three variables showed a normal distribution at each gestational week. Gestation specific reference standards for each variable were constructed for male and female infants separately. Comparison with the 1982-1986 cohort showed a significant secular trend to increased birth weight. The trend was small, but significant, for crown heel length and head circumference. CONCLUSION: These growth standards will provide useful references for the care of newborns of ethnic Chinese origin. These standards, especially that for birth weight, should be updated regularly. PMID- 12719399 TI - Absence of leukaemic fusion gene transcripts in preterm infants exposed to diagnostic x rays. AB - BACKGROUND: Childhood leukaemias express novel, clonotypic fusion genes that may already be present at birth before the clinical manifestation of leukaemia. Exposure of the fetus to diagnostic x rays is reported to increase the risk of childhood leukaemia, and may do so by generating leukaemic fusion genes. Advances in neonatal medicine in the past decade that have extended the limits of viability of preterm babies down to 23 weeks of gestation have resulted in the increased use of diagnostic x rays to monitor neonatal progress. AIM: To investigate whether exposure of very preterm infants to diagnostic x rays in the neonatal period leads to the development of leukaemic fusion genes. METHODS: Peripheral blood samples were collected at birth from very preterm infants (23-30 weeks gestation) and following exposure to diagnostic x rays at intervals of two weeks, until discharge. Cord blood samples from normal full term infants served as controls. Total RNA was extracted from the blood and the expression of the fusion genes TEL-AML1, MLL-AF4, and BCR-ABL, characteristic of three subtypes of childhood leukaemia, was investigated in the preterm and full term infant samples using a nested reverse transcriptase polymerase chain reaction method. Serial pre and post-x ray samples from 42 preterm babies, pre-x ray samples from an additional 46 preterm infants, and cord blood samples from 100 normal full term infants were screened for fusion gene transcripts. RESULTS: No leukaemic fusion gene transcripts were detected in preterm infants following exposure to diagnostic x rays. A BCR-ABL transcript was identified in a single preterm infant prior to x ray exposure. TEL-AML1 transcripts were detected in cord blood samples from two full term infants. MLL-AF4 transcripts were not detected in any of the pre- or full term infants tested. CONCLUSIONS: Exposure of the preterm infants to x rays in this small series and at the doses used for diagnostic purposes did not induce leukaemic fusion gene expression, but we cannot exclude the possibility that a small proportion of preterm infants may be unusually sensitive to x rays. PMID- 12719402 TI - Hugh Downman, MD (1740-1809) of Exeter and his poem on infant care. AB - Hugh Downman is best remembered for his poem on the care of infants in which he stressed the importance of breast feeding and proper examination. PMID- 12719401 TI - Sporadic case of fatal encephalopathy with neonatal onset associated with a T158M missense mutation in MECP2. AB - The case of a male infant with neonatal encephalopathy and a de novo MECP2 mutation is reported. The presenting phenotype of decelerating head growth, spasticity, scoliosis, and central respiratory disturbance raised suspicions of the diagnosis. Neonatal encephalopathy in males is part of a spectrum of disorders caused by MECP2 dysfunction. PMID- 12719400 TI - Decreased neutrophil apoptosis in tracheal fluids of preterm infants at risk of chronic lung disease. AB - OBJECTIVE: To investigate the hypothesis that preterm infants who are more susceptible to lung damage have decreased neutrophil apoptosis, and to explore its relation to interleukin 10 (IL10) concentration. DESIGN: Prospective cohort design. PATIENTS: One hundred tracheal fluid specimens from 50 week-1 ventilated infants were examined for IL10 (by enzyme linked immunosorbent assay) and neutrophil apoptosis (by light microscopy). RESULTS: Neutrophil apoptosis was absent or less than 0.22% (median 0%) in the 11 infants with chronic lung disease (CLD) (24-31 weeks gestation) during the first 4 days of life. This was significantly lower than that of the 20 preterm infants without CLD (27-31 weeks gestation; median 0.47%, range 0-1.25%) and 19 term infants (median 0.5%, range 0 2.25%). There was an increase in apoptosis in infants with CLD (median 0.44%, p = 0.046) during days 5-7. Few infants without CLD were intubated beyond 4 days. Median apoptosis on days 5-7 was 0.26% and 2.78% for non-CLD preterm and term infants, but differences were not significant. IL10 concentration in tracheal fluid of infants with CLD was less than 5 pg/ml. None of the infants with IL10 greater than 5 pg/ml developed CLD. The range of IL10 concentrations in tracheal fluid from infants without CLD was wide (0-938 pg/ml). There was no apparent correlation between IL10 levels and percentage neutrophil apoptosis in infants without CLD. CONCLUSION: Preterm infants with low levels of IL10 and neutrophil apoptosis may be predisposed to disordered lung repair. Further studies into the method of disposal of senescent neutrophils within preterm lungs are required. PMID- 12719403 TI - Reducing antibiotic use on the neonatal unit by improving communication of blood culture results: a completed audit cycle. PMID- 12719404 TI - Amiodarone and breast feeding. PMID- 12719405 TI - Preventing hypothermia at birth in preterm babies: at a cost of overheating some? PMID- 12719406 TI - Technique for insertion of percutaneous central venous catheters in the newborn period. PMID- 12719407 TI - Swaddling and heat loss. PMID- 12719408 TI - Progressive ventricular dilatation (PVD) over the past 22 years. PMID- 12719411 TI - Do we need to assess the thyroid function in the infants of mothers with Hashimoto's thyroiditis? PMID- 12719409 TI - Umbilical granulomas: a randomised controlled trial. PMID- 12719412 TI - The mechanism of gamma-secretase: multiple inhibitor binding sites for transition state analogs and small molecule inhibitors. AB - Transition state analogs pepstatin methylester (PME) and L685458 have been shown to inhibit gamma-secretase non-competitively (Tian, G., Sobotka-Briner, C., Zysk, J., Liu, X., Birr, C., Sylvester, M. A., Edwards, P. D., Scott, C. W., and Greenberg, B. D. (2002) J. Biol. Chem. 277, 31499-31505). This unusual kinetics suggests physical separation of the sites for substrate binding and catalysis with binding of the transition state analogs to the catalytic site and not to the substrate binding site. Methods of inhibitor cross-competition kinetics and competition ligand binding were utilized to address whether non-transition state small molecule inhibitors, which also display non-competitive inhibition of gamma secretase, inhibit the enzyme by binding to the catalytic site as well. Inhibitor cross-competition kinetics indicated competitive binding between the transition state analogs PME and L685458 and between small molecules arylsulfonamides and benzodiazepines, but non-competitive binding between the transition state analogs and the small molecule inhibitors. These results were indicative of two inhibitor binding sites, one for transition state analogs and the other for non-transition state small molecule inhibitors. The presence of two inhibitor binding sites for two different classes of inhibitors was corroborated by results from competition ligand binding using [3H]L685458 as the radioligand. Although L685458 and PME displaced the radioligand at the same concentrations as for enzyme inhibition, arylsulfonamides and benzodiazepines did not displace the radioligand at their Ki values, a result consistent with the presence of two inhibitor binding sites. These findings provide useful insights into the catalytic and regulatory mechanisms of gamma-secretase that may facilitate the design of novel gamma secretase inhibitors. PMID- 12719413 TI - p38 mitogen-activated protein kinase regulates oscillation of chick pineal circadian clock. AB - Extracellular signal-regulated kinase (ERK) and p38 are members of the mitogen activated protein kinase (MAPK) family, and in some cases these kinases serve for closely related cellular functions within a cell. In a wide range of animal clock structures, ERK plays an important role in the circadian time-keeping mechanism. Here we found that immunoreactivity to p38 protein was uniformly distributed among cells in the chick pineal gland. On the other hand, a constant level of activated p38 was detected over the day, predominantly in the follicular and parafollicular pinealocytes that are potential circadian clock-containing cells. Chronic application of SB203580, a selective and reversible inhibitor of p38, to the cultured chick pineal cells markedly lengthened the period of the circadian rhythm of the melatonin release (up to 28.7 h). Noticeably, despite no significant temporal change of activated p38 level, a 4-h pulse treatment with SB203580 delayed the phase of the rhythm only when delivered during the subjective day. These results indicate a time-of-day-specific role of continuously activated p38 in the period length regulation of the chick pineal clock and suggest temporally separated regulation of the clock by two MAPKs, nighttime-activated ERK and daytime-working p38. PMID- 12719414 TI - Changing the conformation state of cytochrome b558 initiates NADPH oxidase activation: MRP8/MRP14 regulation. AB - Phagocyte NADPH oxidase generates O2. for defense mechanisms and cellular signaling. Myeloid-related proteins MRP8 and MRP14 of the S100 family are EF-hand calcium-binding proteins. MRP8 and MRP14 were co-isolated from neutrophils on an anti-p47phox matrix with oxidase cytosolic factors and identified by mass spectrometry. MRP8 and MRP14 are absent from Epstein-Barr virus-immortalized B lymphocytes, and, coincidentally, these cells display weak oxidase activity compared with neutrophils. MRP8/MRP14 that was purified from neutrophils enhanced oxidase turnover of B cells in vitro, suggesting that MRP8/MRP14 is involved in the activation process. This was confirmed ex vivo by co-transfection of Epstein Barr virus-transformed B lymphocytes with genes encoding MRP8 and MRP14. In a semi-recombinant cell-free assay, recombinant MRP8/MRP14 increased the affinity of p67phox for cytochrome b558 synergistically with p47phox. Moreover, MRP8/MRP14 initiated oxidase activation on its own, through a calcium-dependent specific interaction with cytochrome b558 as shown by atomic force microscopy and a structure-function relationship investigation. The data suggest that the change of conformation in cytochrome b558, which initiates the electron transfer, can be mediated by effectors other than oxidase cytosolic factors p67phox and p47phox. Moreover, MRP8/MRP14 dimer behaves as a positive mediator of phagocyte NADPH oxidase regulation. PMID- 12719415 TI - Covalent and non-covalent interactions of betaig-h3 with collagen VI. Beta ig-h3 is covalently attached to the amino-terminal region of collagen VI in tissue microfibrils. AB - Transforming growth factor-beta induced gene-h3 (betaig-h3) was found to co purify with collagen VI microfibrils, extracted from developing fetal ligament, after equilibrium density gradient centrifugation under both nondenaturing and denaturing conditions. Analysis of the collagen VI fraction from the non denaturing gradient by gel electrophoresis under non-reducing conditions revealed the present of a single high molecular weight band that immunostained for both collagen VI and betaig-h3. When the fraction was analyzed under reducing conditions, collagen VI alpha chains and betaig-h3 were the only species evident. The results indicated that betaig-h3 is associated with collagen VI in tissues by reducible covalent bonding, presumably disulfide bridges. Rotary shadowing and immunogold staining of the collagen VI microfibrils and isolated tetramers indicated that betaig-h3 was specifically and periodically associated with the double-beaded region of many of the microfibrils and that this covalent binding site was located in or near the amino-terminal globular domain of the collagen VI molecule. Using solid phase and co-immunoprecipitation assays, recombinant betaig h3 was found to bind both native and pepsin-treated collagen VI but not individual pepsin-collagen VI alpha chains. Blocking experiments indicated that the major in vitro betaig-h3 binding site was located in the pepsin-resistant region of collagen VI. In contrast to the tissue situation, the in vitro interaction had the characteristics of a reversible non-covalent interaction, and the Kd was measured as 1.63 x 10(-8) m. Rotary shadowing of immunogold-labeled complexes of recombinant betaig-h3 and pepsin-collagen VI indicated that the in vitro betaig-h3 binding site was located close to the amino-terminal end of the collagen VI triple helix. The evidence indicates that collagen VI may contain distinct covalent and non-covalent binding sites for betaig-h3, although the possibility that both interactions use the same binding region is discussed. Overall the study supports the concept that betaig-h3 is extensively associated with collagen VI in some tissues and that it plays an important modulating role in collagen VI microfibril function. PMID- 12719416 TI - Intervertebral disc collagen. Usage of the short form of the alpha1(IX) chain in bovine nucleus pulposus. AB - Nucleus pulposus, the central zone of the intervertebral disc, is gel-like and has a similar collagen phenotype to that of hyaline cartilage. Amino-terminal protein sequence analysis of the alpha1(IX)COL3 domain purified from bovine nucleus pulposus gave a different sequence to that of the long alpha1(IX) transcript expressed in hyaline cartilage and matched the predicted sequence of short alpha1(IX). The findings indicate that the matrix of bovine nucleus pulposus contains only the short form of alpha1(IX) that lacks the NC4 domain. The sequence encoded by exon 7, predicted from human COL9A1, is absent from both short and long forms of alpha1(IX) from bovine nucleus pulposus and articular cartilage. A structural analysis of the cross-linking sites occupied in type IX collagen from nucleus pulposus showed that usage of the short alpha1(IX) transcript in disc tissue had no apparent effect on cross-linking behavior. As in cartilage, type IX collagen of nucleus pulposus was heavily cross-linked to type II collagen and to other molecules of type IX collagen with a similar site occupancy. PMID- 12719417 TI - Streptococcus pneumoniae hyaluronate lyase contains two non-cooperative independent folding/unfolding structural domains: characterization of functional domain and inhibitors of enzyme. AB - Hyaluronate lyase contributes directly to bacterial invasion by degrading hyaluronan, the major component of host extracellular matrix of connective tissues. Streptococcus pneumoniae hyaluronate lyase (SpnHL) is built from two structural domains that interact through interface residues, in addition to being connected by a peptide linker. For the first time we demonstrate that the N- and C-terminal domains of SpnHL fold/unfold independent of each other suggesting the absence of any significant cooperative interactions between them. The C-terminal domain of SpnHL is less stable than the N-terminal domain against thermal and guanidine hydrochloride denaturation. The intact N-terminal domain was purified after limited proteolysis of SpnHL under conditions where only the C-terminal domain was unfolded. Isolated N-terminal domain of SpnHL had similar thermal stability as when present in the native enzyme and was found to be enzymatically active demonstrating that it is capable of carrying out enzymatic reaction on its own. Functional studies demonstrated that guanidine hydrochloride, guanidine isothiocyanate, l-arginine methyl ester, and l-arginine inhibit the enzymatic activity of SpnHL at very low concentrations. This provides a lead for new chemical entities that can be exploited for designing effective inhibitors of SpnHL. PMID- 12719419 TI - DNA-protein cross-link formation mediated by oxanine. A novel genotoxic mechanism of nitric oxide-induced DNA damage. AB - Chronic inflammation is a risk factor for many human cancers, and nitric oxide (NO) produced in inflamed tissues has been proposed to cause DNA damage via nitrosation or oxidation of base moieties. Thus, NO-induced DNA damage could be relevant to carcinogenesis associated with chronic inflammation. In this report, we report a novel genotoxic mechanism of NO that involves DNA-protein cross-links (DPCs) induced by oxanine (Oxa), a major NO-induced guanine lesion. When a duplex DNA containing Oxa at the site-specific position was incubated with DNA-binding proteins such as histone, high mobility group (HMG) protein, and DNA glycosylases, DPCs were formed between Oxa and protein. The rate of DPC formation with DNA glycosylases was approximately two orders of magnitude higher than that with histone and HMG protein. Analysis of the reactivity of individual amino acids to Oxa suggested that DPC formation occurred between Oxa and side chains of lysine or arginine in the protein. A HeLa cell extract also gave rise to two major DPCs when incubated with DNA-containing Oxa. These results reveal a dual aspect of Oxa as causal damage of DPC formation and as a suicide substrate of DNA repair enzymes, both of which could pose a threat to the genetic and structural integrity of DNA, hence potentially leading to carcinogenesis. PMID- 12719418 TI - Vipera lebetina venom contains two disintegrins inhibiting laminin-binding beta1 integrins. AB - To explain the myotoxic effects of snake venoms, we searched for inhibitors of alpha7beta1 integrin, the major laminin-binding integrin in skeletal muscle. We discovered two inhibitors in the venom of Vipera lebetina. One of them, lebein-1 (known as lebein), has already been proposed to be a disintegrin because of its RGD-containing primary sequence. The other, lebein-2, is a novel protein that also interacts firmly with alpha3beta1, alpha6beta1, and alpha7beta1 integrins, but not with the collagen-binding alpha1beta1 and alpha2beta1 integrins. Ligand binding of laminin-recognizing beta1 integrins was efficiently blocked by both lebein-1 and lebein-2. In cell attachment assays, lebein-1 and lebein-2 inhibited myoblast attachment not only to laminin, but also to fibronectin. However, neither lebein-1 nor lebein-2 interacted with alpha7beta1 integrin in an RGD dependent manner, similar to the interaction of the laminin with alpha7beta1 integrin. Identical divalent cation dependence of integrin binding to laminin and to either of the two inhibitors and their mutually exclusive binding suggest that both lebein-1 and lebein-2 interact with the ligand-binding site of laminin binding beta1 integrins by mimicking the yet unknown integrin-binding structure of laminins. Like lebein-1, lebein-2 is a soluble heterodimeric disintegrin of low molecular mass. Together with membrane-bound ADAM-2 and ADAM-9, the two inhibitors seem to form a small group of disintegrins that can bind to laminin binding beta1 integrins. Because of their inhibitory capability both in vitro and in vivo, lebein-1 and lebein-2 may be valuable tools in influencing laminin induced, integrin-mediated cell functions such as cell anchorage, migration, and mechanical force transduction on laminin-rich basement membranes. PMID- 12719420 TI - Biglycan, a nitric oxide-regulated gene, affects adhesion, growth, and survival of mesangial cells. AB - During glomerular inflammation mesangial cells are the major source and target of nitric oxide that pro-foundly influences proliferation, adhesion, and death of mesangial cells. The effect of nitric oxide on the mRNA expression pattern of cultured rat mesangial cells was therefore investigated by RNA-arbitrarily-primed polymerase chain reaction. Employing this approach, biglycan expression turned out to be down-regulated time- and dose-dependently either by interleukin-1beta stimulated endogenous nitric oxide production or by direct application of the exogenous nitric oxide donor, diethylenetriamine nitric oxide. There was a corresponding decline in the rate of biglycan biosynthesis and in the steady state level of this proteoglycan. In vivo, in a model of mesangioproliferative glomerulonephritis up-regulation of inducible nitric-oxide synthase mRNA was associated with reduced expression of biglycan in isolated glomeruli. Biglycan expression could be normalized, both in vitro and in vivo, by using a specific inhibitor of the inducible nitric-oxide synthase, l-N6-(l-iminoethyl)-l-lysine dihydrochloride. Further studies showed that biglycan inhibited cell adhesion on type I collagen and fibronectin because of its binding to these substrates. More importantly, biglycan protected mesangial cells from apoptosis by decreasing caspase-3 activity, and it counteracted the proliferative effects of platelet derived growth factor-BB. These findings indicate a signaling role of biglycan and describe a novel pathomechanism by which nitric oxide modulates the course of renal glomerular disease through regulation of biglycan expression. PMID- 12719421 TI - Structural flexibility of multifunctional HABP1 may be important for regulating its binding to different ligands. AB - Hyaluronan-binding protein 1 (HABP1)/p32/gC1qR was characterized as a highly acidic and oligomeric protein, which binds to different ligands like hyaluronan, C1q, and mannosylated albumin. It exists as trimer in high ionic and reducing conditions as shown by crystal structure. In the present study, we have examined the structural changes of HABP1 under a wide range of ionic environments. HABP1 exhibits structural plasticity, which is influenced by the ionic environment under in vitro conditions near physiological pH. At low ionic strength HABP1 exists in a highly expanded and loosely held trimeric structure, similar to that of the molten globule-like state, whereas the presence of salt stabilizes the trimeric structure in a more compact fashion. It is likely that the combination of the high net charge asymmetrically distributed along the faces of the molecule and the relatively low intrinsic hydrophobicity of HABP1 result in its expanded structure at neutral pH. Thus, the addition of counter ions in the molecular environment minimizes the intramolecular electrostatic repulsion in HABP1 leading to its stable and compact conformations, which reflect in its differential binding toward different ligands. Whereas the binding of HABP1 toward HA is enhanced on increasing the ionic strength, no significant effect was observed with the two other ligands, C1q and mannosylated albumin. Thus, although HA interacts only with compact HABP1, C1q and mannosylated albumin can bind to loosely held oligomeric HABP1 as well. In other words, structural changes in HABP1 mediated by changes in the ionic environment are responsible for recognizing different ligands. PMID- 12719422 TI - Inhibition of transcription factor Stat5 induces cell death of human prostate cancer cells. AB - Identifying regulators of prostate cancer cell survival may lead to new therapeutic strategies for prostate cancer. We now report prevalent activation of transcription factor Stat5 in human prostate cancer and provide novel evidence that blocking activation of Stat5 in human prostate cancer cells leads to extensive cell death. Specifically, Stat5 was activated in 65% of human prostate cancer specimens examined based on nuclear location of tyrosine phosphorylated Stat5. Adenoviral gene delivery of a dominant-negative Stat5 mutant (DNStat5), but not wild-type Stat5, induced cell death of both the androgen-independent human prostate cancer cell line CWR22Rv and the androgen-sensitive LnCap cell line. Endogenous Stat5 was active in both CWR22Rv and LnCap cells. In contrast, only low levels of inactive Stat5 proteins were detected in the PC-3 cell line, which correlated with resistance to DNStat5-induced cell death. In CWR22Rv and LnCap cells, inhibition of Stat5 by expression of DNStat5 induced apoptotic cell death as judged from morphological changes, DNA fragmentation, and caspase-3 activation with evidence of a caspase-9-dependent mechanism. We propose that blocking Stat5 function may represent a novel therapeutic approach for prostate cancer. PMID- 12719423 TI - The inhibition of early N-glycan processing targets TRP-2 to degradation in B16 melanoma cells. AB - Tyrosinase-related protein-2 (TRP-2) is a DOPAchrome tautomerase catalyzing a distal step in the melanin synthesis pathway. Similar to the other two melanogenic enzymes belonging to the TRP gene family, tyrosinase and TRP-1, TRP-2 is expressed in melanocytes and melanoma cells. Despite the increasing evidence of its efficiency as a melanoma antigen, little is known about the maturation and intracellular trafficking of TRP-2. Here we show that TRP-2 is mainly distributed in the TGN of melanoma cells instead of being confined solely to melanosomes. This, together with the plasma membrane occasional localization observed by immunofluorescence, suggest the TRP-2 participation in a recycling pathway, which could include or not the melanosomes. Using pulse-chase experiments we show that the TRP-2 polypeptide folds in the endoplasmic reticulum (ER) in the presence of calnexin, until it reaches a dithiothreitol-resistant conformation enabling its ER exit to the Golgi. If N-glycosylation inhibitors prevent the association with calnexin, the TRP-2 nascent chain undergoes an accelerated degradation process. This process is delayed in the presence of proteasomal inhibitors, indicating that the misfolded chain is retro-translocated from the ER into the cytosol and degraded in proteasomes. This is a rare example in which calnexin although indispensable for the nascent chain folding is not required for its targeting to degradation. Therefore TRP-2 may prove to be a good model to document the calnexin-independent retro-translocation process of proteasomally degraded proteins. Clearly, TRP-2 has a distinct maturation pathway from tyrosinase and TRP-1 and possibly a second regulatory function within the cell. PMID- 12719424 TI - Rh-RhAG/ankyrin-R, a new interaction site between the membrane bilayer and the red cell skeleton, is impaired by Rh(null)-associated mutation. AB - Several studies suggest that the Rh complex represents a major interaction site between the membrane lipid bilayer and the red cell skeleton, but little is known about the molecular basis of this interaction. We report here that ankyrin-R is capable of interacting directly with the C-terminal cytoplasmic domain of Rh and RhAG polypeptides. We first show that the primary defect of ankyrin-R in normoblastosis (nb/nb) spherocytosis mutant mice is associated with a sharp reduction of RhAG and Rh polypeptides. Secondly, our flow cytometric analysis of the Triton X-100 extractability of recombinant fusion proteins expressed in erythroleukemic cell lines suggests that the C-terminal cytoplasmic domains of Rh and RhAG are sufficient to mediate interaction with the erythroid membrane skeleton. Using the yeast two-hybrid system, we demonstrate a direct interaction between the cytoplasmic tails of Rh and RhAG and the second repeat domain (D2) of ankyrin-R. This finding is supported by the demonstration that the substitution of Asp-399 in the cytoplasmic tail of RhAG, a mutation associated with the deficiency of the Rh complex in one Rhnull patient, totally impaired interaction with domain D2 of ankyrin-R. These results identify the Rh/RhAG-ankyrin complex as a new interaction site between the red cell membrane and the spectrin-based skeleton, the disruption of which might result in the stomato-spherocytosis typical of Rhnull red cells. PMID- 12719425 TI - A reaction center-light-harvesting 1 complex (RC-LH1) from a Rhodospirillum rubrum mutant with altered esterifying pigments: characterization by optical spectroscopy and cryo-electron microscopy. AB - Introduction of the bchP gene from Rhodobacter sphaeroides encoding geranylgeranyl reductase into Rhodospirillum rubrum alters the esterification of the bacteriochlorophylls so that phytol is used instead of geranylgeraniol. The resulting transconjugant strain of Rs. rubrum grows photosynthetically, showing that phytolated Bchla can substitute for the native pigment in both the reaction center (RC) and the light-harvesting 1 (LH1) complexes. This genetic manipulation perturbs the native carotenoid biosynthetic pathway; several biosynthetic intermediates are assembled into the core complex and are capable of energy transfer to the bacteriochlorophylls. RC-LH1 complexes containing phytolated Bchla were analyzed by low temperature absorption and fluorescence spectroscopy and circular dichroism. These show that phytolated Bchls can assemble in vivo into the photosynthetic apparatus of Rs. rubrum and that the newly introduced phytol tail provokes small perturbations to the Bchls within their binding sites in the LH1 complex. The RC-LH1 core complex was purified from membranes and reconstituted into well ordered two-dimensional crystals with a p4212 space group. A projection map calculated to 9 A shows clearly that the LH1 ring from the mutant is composed of 16 subunits that surround the reaction center and that the diameter of this complex is in close agreement with that of the wild-type LH1 complex. PMID- 12719426 TI - Biochemical analysis of the yeast condensin Smc2/4 complex: an ATPase that promotes knotting of circular DNA. AB - To better understand the contributions that the structural maintenance of chromosome proteins (SMCs) make to condensin activity, we have tested a number of biochemical, biophysical, and DNA-associated attributes of the Smc2p-Smc4p pair from budding yeast. Smc2p and Smc4p form a stable heterodimer, the "Smc2/4 complex," which upon analysis by sedimentation equilibrium appears to reversibly self-associate to form heterotetramers. Individually, neither Smc2p nor Smc4p hydrolyzes ATP; however, ATPase activity is recovered by equal molar mixing of both purified proteins. Hydrolysis activity is unaffected by the presence of DNA. Smc2/4 binds both linearized and circular plasmids, and the binding appears to be independent of adenylate nucleotide. High mole ratios of Smc2/4 to plasmid promote a geometric change in circular DNA that can be trapped as knots by type II topoisomerases but not as supercoils by a type I topoisomerase. Binding titration analyses reveal that two Smc2/4-DNA-bound states exist, one disrupted by and one resistant to salt challenge. Competition-displacement experiments show that Smc2/4-DNA-bound species formed at even high protein to DNA mole ratios remain reversible. Surprisingly, only linear and supercoiled DNA, not nicked circular DNA, can completely displace Smc2/4 prebound to a labeled, nicked circular DNA. To explain this geometry-dependent competition, we present two models of DNA binding by SMCs in which two DNA duplexes are captured within the inter-coil space of an Smc2/4 heterodimer. Based on these models, we propose a DNA displacement mechanism to explain how differences in geometry could affect the competitive potential of DNA. PMID- 12719427 TI - Biochemical and structural analysis of the molybdenum cofactor biosynthesis protein MobA. AB - Molybdopterin guanine dinucleotide (MGD) is the form of the molybdenum cofactor that is required for the activity of most bacterial molybdoenzymes. MGD is synthesized from molybdopterin (MPT) and GTP in a reaction catalyzed by the MobA protein. Here we report that wild type MobA can be copurified along with bound MPT and MGD, demonstrating a tight binding of both its substrate and product. To study structure-function relationships, we have constructed a number of site specific mutations of the most highly conserved amino acid residues of the MobA protein family. Variant MobA proteins were characterized for their ability to support the synthesis of active molybdenum enzymes, to bind MPT and MGD, to interact with the molybdenum cofactor biosynthesis proteins MobB and MoeA. They were also characterized by x-ray structural analysis. Our results suggest an essential role for glycine 15 of MobA, either for GTP binding and/or catalysis, and an involvement of glycine 82 in the stabilization of the product-bound form of the enzyme. Surprisingly, the individual and double substitution of asparagines 180 and 182 to aspartate did not affect MPT binding, catalysis, and product stabilization. PMID- 12719429 TI - Amino acids of the bacterial toxin SopE involved in G nucleotide exchange on Cdc42. AB - RhoGTPases are central switches in all eukaryotic cells. There are at least two known families of guanine nucleotide exchange factors that can activate RhoGTPases: the Dbl-like eukaryotic G nucleotide exchange factors and the SopE like toxins of pathogenic bacteria, which are injected into host cells to manipulate signaling. Both families have strikingly different sequences, structures, and catalytic core elements. This suggests that they have emerged by convergent evolution. Nevertheless, both families of G nucleotide exchange factors also share some similarities: (a) both rearrange the G nucleotide binding site of RhoGTPases into virtually identical conformations, and (b) two SopE residues (Gln-109SopE and Asp-124SopE) engage Cdc42 in a similar way as equivalent residues of Dbl-like G nucleotide exchange factors (i.e. Asn-810Dbs and Glu-639Dbs). The functional importance of these observations has remained unclear. Here, we have analyzed the effect of amino acid substitutions at selected SopE residues implicated in catalysis (Asp-124SopE, Gln-109SopE, Asp 103SopE, Lys-198SopE, and Gly-168SopE) on in vitro catalysis of G nucleotide release from Cdc42 and on in vivo activity. Substitutions at Asp-124SopE, Gln 109SopE, and Gly-168SopE severely reduced the SopE activity. Slight defects were observed with Asp-103SopE variants, whereas Lys-198SopE was not found to be required in vitro or in vivo. Our results demonstrate that G nucleotide exchange by SopE involves both catalytic elements unique to the SopE family (i.e. 166GAGA169 loop, Asp-103SopE) and amino acid contacts resembling those of key residues of Dbl-like guanine nucleotide exchange factors. Therefore, besides all of the differences, the catalytic mechanisms of the SopE and the Dbl families share some key functional aspects. PMID- 12719428 TI - NPC1 and NPC2 regulate cellular cholesterol homeostasis through generation of low density lipoprotein cholesterol-derived oxysterols. AB - Mutations in the Niemann-Pick disease genes cause lysosomal cholesterol accumulation and impaired low density lipoprotein (LDL) cholesterol esterification. These findings have been attributed to a block in cholesterol movement from lysosomes to the site of the sterol regulatory machinery. In this study we show that Niemann-Pick type C1 (NPC1) and Niemann-Pick type C2 (NPC2) mutants have increased cellular cholesterol, yet they are unable to suppress LDL receptor activity and cholesterol biosynthesis. Cholesterol overload in both NPC1 and NPC2 mutants results from the failure of LDL cholesterol tobothsuppresssterolregulatoryelement-bindingprotein-dependent gene expression and promote liver X receptor-mediated responses. However, the severity of the defect in regulation of sterol homeostasis does not correlate with endoplasmic reticulum cholesterol levels, but rather with the degree to which NPC mutant fibroblasts fail to appropriately generate 25-hydroxycholesterol and 27 hydroxycholesterol in response to LDL cholesterol. Moreover, we demonstrate that treatment with oxysterols reduces cholesterol in NPC mutants and is able to correct the NPC1I1061T phenotype, the most prevalent NPC1 disease genotype. Our findings support a role for NPC1 and NPC2 in the regulation of sterol homeostasis through generation of LDL cholesterol-derived oxysterols and have important implications for the treatment of NPC disease. PMID- 12719430 TI - Polymerization of mycobacterial arabinogalactan and ligation to peptidoglycan. AB - The cell wall of Mycobacterium spp. consists predominately of arabinogalactan chains linked at the reducing ends to peptidoglycan via a P-GlcNAc-(alpha1-3)-Rha linkage unit (LU) and esterified to a variety of mycolic acids at the nonreducing ends. Several aspects of the biosynthesis of this complex have been defined, including the initial formation of the LU on a polyprenyl phosphate (Pol-P) molecule followed by the sequential addition of galactofuranosyl (Galf) units to generate Pol-P-P-LU-(Galf)1,2,3, etc. and Pol-P-P-LU-galactan, catalyzed by a bifunctional galactosyltransferase (Rv3808c) capable of adding alternating 5- and 6-linked Galf units. By applying cell-free extracts of Mycobacterium smegmatis, containing cell wall and membrane fragments, and differential labeling with UDP [14C]Galp and recombinant UDP-Galp mutase as the source of [14C]Galf for galactan biosynthesis and 5-P-[14C]ribosyl-P-P as a donor of [14C]Araf for arabinan synthesis, we now demonstrate sequential synthesis of the simpler Pol-P-P-LU (Galf)n glycolipid intermediates followed by the Pol-P-P-LU-arabinogalactan and, finally, ligation of the P-LU-arabinogalactan to peptidoglycan. This first time demonstration of in vitro ligation of newly synthesized P-LU-arabinogalactan to newly synthesized peptidoglycan is a necessary forerunner to defining the genetics and enzymology of cell wall polymer-peptidoglycan ligation in Mycobacterium spp. and examining this step as a target for new antibacterial drugs. PMID- 12719431 TI - Mitotic and stress-induced phosphorylation of HsPI3K-C2alpha targets the protein for degradation. AB - Activation of the phosphoinositide 3-kinases (PI 3-kinases) has been implicated in multiple cellular responses such as proliferation and survival, membrane and cytoskeletal reorganization, and intracellular vesicular trafficking. The activities and subcellular localization of PI 3-kinases were shown to be regulated by phosphorylation. Previously we demonstrated that class II HsPIK3 C2alpha becomes phosphorylated upon inhibition of RNA pol II-dependent transcription (Didichenko, S. A., and Thelen, M. (2001) J. Biol. Chem. 276, 48135 48142). In this study we investigated cell cycle-dependent and genotoxic stress induced phosphorylation of HsPIK3-C2alpha. We find that the kinase becomes phosphorylated upon exposure of cells to UV irradiation and in proliferating cells at the G2/M transition of the cell cycle. Stress-dependent and mitotic phosphorylation of HsPIK3-C2alpha occurs on the same serine residue (Ser259) within a recognition motif for proline-directed kinases. Mitotic phosphorylation of HsPIK3-C2alpha can be attributed to Cdc2 activity, and stress-induced phosphorylation of HsPIK3-C2alpha is mediated by JNK/SAPK. The protein level of HsPIK3-C2alpha is regulated by proteolysis in a cell cycle-dependent manner and in response of cells to stress. Phosphorylation appears to be a prerequisite for proteasome-dependent degradation of HsPIK3-C2alpha and may therefore contribute indirectly to the regulation of the activity of the kinase. PMID- 12719432 TI - Functional complementation between a novel mammalian polygenic transport complex and an evolutionarily ancient organic solute transporter, OSTalpha-OSTbeta. AB - These studies identify an organic solute transporter (OST) that is generated when two novel gene products are co-expressed, namely human OSTalpha and OSTbeta or mouse OSTalpha and OSTbeta. The results also demonstrate that the mammalian proteins are functionally complemented by evolutionarily divergent Ostalpha Ostbeta proteins recently identified in the little skate, Raja erinacea, even though the latter exhibit only 25-41% predicted amino acid identity with the mammalian proteins. Human, mouse, and skate OSTalpha proteins are predicted to contain seven transmembrane helices, whereas the OSTbeta sequences are predicted to have a single transmembrane helix. Human OSTalpha-OSTbeta and mouse Ostalpha Ostbeta cDNAs were cloned from liver mRNA, sequenced, expressed in Xenopus laevis oocytes, and tested for their ability to functionally complement the corresponding skate proteins by measuring transport of [3H]estrone 3-sulfate. None of the proteins elicited a transport signal when expressed individually in oocytes; however, all nine OSTalpha-OSTbeta combinations (i.e. OSTalpha-OSTbeta pairs from human, mouse, or skate) generated robust estrone 3-sulfate transport activity. Transport was sodium-independent, saturable, and inhibited by other steroids and anionic drugs. Human and mouse OSTalpha-OSTbeta also were able to mediate transport of taurocholate, digoxin, and prostaglandin E2 but not of estradiol 17beta-d-glucuronide or p-aminohippurate. OSTalpha and OSTbeta were able to reach the oocyte plasma membrane when expressed either individually or in pairs, indicating that co-expression is not required for proper membrane targeting. Interestingly, OSTalpha and OSTbeta mRNAs were highly expressed and widely distributed in human tissues, with the highest levels occurring in the testis, colon, liver, small intestine, kidney, ovary, and adrenal gland. PMID- 12719433 TI - Alpha-Synuclein is degraded by both autophagy and the proteasome. AB - Parkinson's disease (PD) is characterized by the loss of dopaminergic neurons in the substantia nigra and the formation of aggregates (Lewy bodies) in neurons. alpha-Synuclein is the major protein in Lewy bodies and rare mutations in alpha synuclein cause early-onset PD. Consequently, alpha-synuclein is implicated in the pathogenesis of PD. Here, we have investigated the degradation pathways of alpha-synuclein, using a stable inducible PC12 cell model, where the expression of exogenous human wild-type, A30P, or A53T alpha-synuclein can be switched on and off. We have used a panel of inhibitors/stimulators of autophagy and proteasome function and followed alpha-synuclein degradation in these cells. We found that not only is alpha-synuclein degraded by the proteasome, but it is also degraded by autophagy. A role for autophagy was further supported by the presence of alpha-synuclein in organelles with the ultrastructural features of autophagic vesicles. Since rapamycin, a stimulator of autophagy, increased clearance of alpha-synuclein, it merits consideration as a potential therapeutic for Parkinsons disease, as it is designed for chronic use in humans. PMID- 12719434 TI - Crystal structure of calcium-free alpha-amylase from Bacillus sp. strain KSM-K38 (AmyK38) and its sodium ion binding sites. AB - The crystal structure of a calcium-free alpha-amylase (AmyK38) from Bacillus sp. strain KSM-K38, which resists chelating reagents and chemical oxidants, has been determined by the molecular replacement method and refined to a crystallographic R-factor of 19.9% (R-free of 23.2%) at 2.13-A resolution. The main chain folding of AmyK38 is almost homologous to that of Bacillus licheniformis alpha-amylase. However, neither a highly conserved calcium ion, which is located at the interface between domains A and B, nor any other calcium ions appear to exist in the AmyK38 molecule, although three sodium ions were found, one of which is located at the position corresponding to that of a highly conserved calcium ion of other alpha-amylases. The existence of these sodium ions was crystallographically confirmed by the structures of three metal-exchanged and mutated enzymes. This is the first case in which the structure of the calcium free alpha-amylase has been determined by crystallography, and it was suggested that these sodium ions, instead of calcium ions, are used to retain the structure and function of AmyK38. PMID- 12719435 TI - Endocytosis of receptor tyrosine kinases is driven by monoubiquitylation, not polyubiquitylation. AB - Growth factors stimulate specific receptor tyrosine kinases, but subsequent receptor endocytosis terminates signaling. The ubiquitin ligase c-Cbl targets epidermal growth factor receptors (EGFRs) to endocytosis by tagging them with multiple ubiquitin molecules. However, the type of ubiquitylation is unknown; whereas polyubiquitin chains signal proteasomal degradation, ubiquitin monomers control other processes. We report that in isolation c-Cbl mediates monoubiquitylation rather than polyubiquitylation of EGFRs. Consistent with the sufficiency of monoubiquitylation, when fused to the tail of EGFR, a single ubiquitin induces receptor endocytosis and degradation in cells. By using receptor and ubiquitin mutants, we infer that c-Cbl attaches a founder monoubiquitin to the kinase domain of EGFR and this is complemented by the conjugation of additional monoubiquitins. Hence, receptor tyrosine kinases are desensitized through conjugation of multiple monoubiquitins, which is distinct from polyubiquitin-dependent proteasomal degradation. PMID- 12719436 TI - Cardiac ischemia activates calcium-independent phospholipase A2beta, precipitating ventricular tachyarrhythmias in transgenic mice: rescue of the lethal electrophysiologic phenotype by mechanism-based inhibition. AB - Murine myocardium contains diminutive amounts of calcium-independent phospholipase A2 (iPLA2) activity (<5% that of human heart), and malignant ventricular tachyarrhythmias are infrequent during acute murine myocardial ischemia. Accordingly we considered the possibility that the mouse was a species specific knockdown of the human pathologic phenotype of ischemiainduced lethal ventricular tachyarrhythmias. Transgenic mice were generated expressing amounts of iPLA2beta activity comparable to that present in human myocardium. Coronary artery occlusion in Langendorff perfused hearts from transgenic mice resulted in a 22-fold increase in fatty acids released into the venous eluent (29.4 nmol/ml in transgenic versus 1.35 nmol/ml of eluent in wild-type mice), a 4-fold increase in lysophosphatidylcholine mass in ischemic zones (4.9 nmol/mg in transgenic versus 1.1 nmol/mg of protein in wild-type mice), and malignant ventricular tachyarrhythmias within minutes of ischemia. Neither normally perfused transgenic nor ischemic wild-type hearts demonstrated these alterations. Pretreatment of Langendorff perfused transgenic hearts with the iPLA2 mechanism-based inhibitor (E)-6-(bromomethylene)-3-(1-naphthalenyl)-2H-tetrahydropyran-2-one (BEL) just minutes prior to induction of ischemia completely ablated fatty acid release and lysolipid accumulation and rescued transgenic hearts from malignant ventricular tachyarrhythmias. Collectively these results demonstrate that ischemia activates iPLA2beta in intact myocardium and that iPLA2beta-mediated hydrolysis of membrane phospholipids can induce lethal malignant ventricular tachyarrhythmias during acute cardiac ischemia. PMID- 12719437 TI - Interaction of activator of G-protein signaling 3 (AGS3) with LKB1, a serine/threonine kinase involved in cell polarity and cell cycle progression: phosphorylation of the G-protein regulatory (GPR) motif as a regulatory mechanism for the interaction of GPR motifs with Gi alpha. AB - Activator of G-protein signaling 3 (AGS3) has a modular domain structure consisting of seven tetratricopeptide repeats (TPRs) and four G-protein regulatory (GPR) motifs. Each GPR motif binds to the alpha subunit of Gi/Go (Gialpha > Goalpha) stabilizing the GDP-bound conformation of Galpha and apparently competing with Gbetagamma for GalphaGDP binding. As an initial approach to identify regulatory mechanisms for AGS3-G-protein interactions, a yeast two-hybrid screen was initiated using the TPR and linker region of AGS3 as bait. This screen identified the serine/threonine kinase LKB1, which is involved in the regulation of cell cycle progression and polarity. Protein interaction assays in mammalian systems using transfected cells or brain lysate indicated the regulated formation of a protein complex consisting of LKB1, AGS3, and G proteins. The interaction between AGS3 and LKB1 was also observed with orthologous proteins in Drosophila where both proteins are involved in cell polarity. LKB1 immunoprecipitates from COS7 cells transfected with LKB1 phosphorylated the GPR domains of AGS3 and the related protein LGN but not the AGS3-TPR domain. GPR domain phosphorylation was completely blocked by a consensus GPR motif peptide, and placement of a phosphate moiety within a consensus GPR motif reduced the ability of the peptide to interact with G-proteins. These data suggest that phosphorylation of GPR domains may be a general mechanism regulating the interaction of GPR-containing proteins with G-proteins. Such a mechanism may be of particular note in regard to localized signal processing in the plasma membrane involving G-protein subunits and/or intracellular functions regulated by heterotrimeric G-proteins that occur independently of a typical G-protein-coupled receptor. PMID- 12719438 TI - Genetic variants of WNK4 in whites and African Americans with hypertension. AB - Human chromosome 17q has been implicated to contain a gene that influences hypertension susceptibility. This region contains the WNK4 gene that causes the mendelian disorder pseudohypoaldosteronism type II, characterized by high potassium levels and hypertension. The goal of this study was to identify genetic variants in all exons of WNK4 in hypertensive individuals and to examine the association of these variants with essential hypertension. Single-nucleotide polymorphims (SNPs) were identified by sequencing the entire coding region in 32 whites and 32 African Americans with hypertension. A single SNP in whites and 8 SNPs in African Americans were genotyped in a larger cohort of whites (165 hypertensives; 91 normotensives) and African Americans (120 hypertensives; 98 normotensives). The frequency of the rare allele differed significantly between hypertensive whites (13.0%) and normotensive whites (7.1%, P=0.040) for the single intronic SNP (bp 1 156 666). This difference remained significant after adjusting for body mass index and sex (P=0.035). Genotypic frequencies differed significantly between hypertensive and normotensive individuals when a dominant model either with (P=0.027) or without (P=0.028) covariate adjustment was assumed. The odds ratio for hypertension was 2.28 for AA or AG individuals vs those with the GG genotype (95% confidence interval, 1.09 to 4.75). No significant differences in allelic or genotypic frequencies were observed in African Americans for any SNPs. The finding in whites is consistent with the hypothesis that polymorphisms in WNK4 influence the risk of hypertension. However, because the associated SNP does not appear to be a functional variant and the limitations of case/control association studies, confirmation of these results in additional cohorts is warranted. PMID- 12719439 TI - Oxidative stress in Dahl salt-sensitive hypertension. AB - The role of oxidative stress in the long-term regulation of arterial pressure, renal hemodynamics, and renal damage was studied in Dahl salt-sensitive rats. Twenty-eight Dahl S/Rapp strain rats, equipped with indwelling arterial and venous catheters, were subjected to a 3-week intravenous infusion of either low Na (0.9 mmol/d) or high Na (20.6 mmol/d) or the superoxide dismutase mimetic, 4 hydroxyl-2,2,6,6-tetramethylpiperidine-1-oxyl (Tempol), at 125 micromol x kg(-1) x h(-1) plus low Na or high Na. After 21 days, mean arterial pressure was 140+/-3 mm Hg in the high-Na group, 118+/-1 mm Hg (P<0.05) in the high-Na/Tempol group, and unchanged in the low-Na/Tempol and low-Na groups. Tempol did not change renal blood flow, glomerular filtration rate, or glomerular cross-sectional area in rats subjected to the high-Na intake but did decrease urinary protein excretion, the percentage of sclerotic glomeruli, and the kidney weight to body weight ratio. In 15 additional Dahl S rats subjected to high or low Na intake for 3 weeks, renal cortical and medullary O2*- release increased significantly in the high-Na group when compared with the low-Na group. Tempol decreased both renal cortical and medullary O2*- release in the high- and low-Na rats, but the decrease in O2*- release was greater in high-Na rats. The data suggest that oxidative stress contributes to Dahl salt-sensitive hypertension and the accompanying renal damage. PMID- 12719440 TI - Effect of treatment with lercanidipine on heart of Cohen-Rosenthal diabetic hypertensive rats. AB - The influence of treatment with the dihydropyridine-type Ca2+ antagonist lercanidipine on heart and coronary microanatomic changes was investigated in spontaneously hypertensive rats, Cohen-diabetic rats, and Cohen-Rosenthal diabetic hypertensive rats. At 12 weeks of age, animals were left untreated (control groups) or were treated for 8 weeks with an oral dose of 3 mg/kg per day of lercanidipine. Wistar-Kyoto rats were used as a normotensive reference group. In spontaneously hypertensive rats and diabetic hypertensive rats, systolic blood pressure was higher in comparison with Wistar-Kyoto rats. Augmented pressure values were decreased by lercanidipine treatment. Systolic blood pressure was slightly higher in Cohen-diabetic rats than in Wistar-Kyoto rats, and this increase was countered by treatment with lercanidipine. In spontaneously hypertensive rats, diabetic rats, and diabetic hypertensive rats, the thickness of left ventricle and cardiocyte area were increased. Focal connective tissue areas and diffuse accumulation of connective tissue were observed in the left ventricle of spontaneously hypertensive and Cohen-diabetic rats, respectively. Pharmacological treatment countered left ventricle thickening and restored cardiocyte area values in subendocardium. An increased thickness of tunica media accompanied by luminal narrowing was found in coronary artery branches of control spontaneously hypertensive and diabetic hypertensive rats. Treatment with lercanidipine countered vascular changes primarily in small-sized coronary arteries. These results indicate that hypertensive, diabetic, and diabetic hypertensive rats undergo cardiac hypertrophy and vascular changes affecting small-sized coronary arteries. Treatment with lercanidipine countered hypertension-related cardiac and coronary changes, suggesting that this dihydropyridine-type Ca2+ antagonist may improve heart and coronary structure in diabetes associated with hypertension. PMID- 12719441 TI - Different effect of antihypertensive drugs on conduit artery endothelial function. AB - To compare the effect of antihypertensive drugs on endothelium-dependent vasodilation in the peripheral conduit arteries of patients with essential hypertension, in a prospective, randomized, parallel group study, endothelial function was assessed in 168 hypertensive patients before and after 6-month treatment with randomly assigned nifedipine GITS (30 to 60 mg, n=28), amlodipine (5 to 10 mg, n=28), atenolol (50 to 100 mg, n=29), nebivolol (5 to 10 mg, n=28), telmisartan (80 to 160 mg, n=29), and perindopril (2 to 4 mg, n=28). If necessary, hydrochlorothiazide (25 mg) was added to each compound. We evaluated brachial artery flow-mediated, endothelium-dependent dilation (high-resolution ultrasound) compared with endothelium-independent response to glyceryl trinitrate (25 microg/s). Brachial artery diameter was measured by automatic computerized analysis. Forty healthy subjects were evaluated as a control group. Oxidative stress production was evaluated by measuring plasma malondialdehyde and plasma lipoperoxides; plasma antioxidant capacity was assessed as ferric-reducing antioxidant power. Hypertensive patients showed a significantly (P<0.01) lower flow-mediated dilation (5.2+/-1.9%) as compared with healthy control subjects (7.1+/-2.6%). Response to glyceryl trinitrate was similar in control subjects and patients. At baseline, blood pressure, diameter, flow-mediated dilation, and response to glyceryl trinitrate were similar in the different treatment groups. All treatments similarly reduced blood pressure, but only perindopril increased flow mediated dilation (from 5.1+/-2 to 6.4+/-2.4%; P<0.01) without modifying the response to glyceryl trinitrate. Perindopril but also telmisartan nifedipine and amlodipine reduced oxidative stress and increased plasma antioxidant capacity. In patients with essential hypertension, ACE inhibitors appear to be the only compounds able to improve conduit artery endothelium-dependent vasodilation. PMID- 12719442 TI - Pressor effect of water drinking in tetraplegic patients may be a spinal reflex. AB - Water drinking elicits a profound sympathetically mediated pressor response in patients with autonomic failure. To further elucidate the mechanism of the response, we assessed the acute effect of drinking water on supine blood pressure and heart rate in 13 tetraplegic patients (12 men, 1 woman; 39+/-4 years of age; body mass index, 25+/-1 kg/m2) with complete spinal cord injury (C2 to C7). Heart rate and finger blood pressure were recorded continuously. Brachial blood pressure was measured every 5 minutes. Baroreflex sensitivity was assessed by the sequence method. Stroke volume was calculated by use of transthoracic bioimpedance. Patients were placed in the supine position with the upper body elevated by 15 degrees. After 30 minutes, supine patients ingested 500 mL of water and the following 60 minutes were monitored. Finger blood pressure at baseline was 123+/-8/65+/-4 mm Hg. Water drinking elicited a pressor response that was apparent within 5 minutes and reached a maximum of 138+/-8/73+/-4 mm Hg after 35 to 40 minutes (P<0.05). Heart rate decreased from 64+/-2 bpm at baseline to 60+/-2 bpm (P<0.001). The mean area under the curve for brachial systolic blood pressure changes differed significantly from zero (364+/-151 mm Hg/min). Total peripheral resistance increased by 15+/-4% (P<0.05). Baroreflex sensitivity increased from 18+/-5 ms/mm Hg at baseline to 23+/-6 ms/mm Hg at 35 minutes after water drinking (P<0.01). Water drinking elicits a pressor response even if the direct connection between brain stem cardiovascular centers and spinal sympathetic neurons is interrupted. This observation might suggest that water drinking activates postganglionic sympathetic neurons either directly or through a spinal reflex mechanism. PMID- 12719443 TI - Functional role of ETB receptors in the renal medulla. AB - Experiments were conducted to determine the influence of ETB receptors in the control of renal medullary function. The acute relation between renal perfusion pressure (RPP) and natriuresis was examined in anesthetized rats treated with the ETB antagonist A-192621 (10 mg/kg IV). In A-192621-treated rats, sodium excretion (UNaV) was 0.4+/-0.1, 0.6+/-0.3, and 2.7+/-0.5 micromol/min at RPP of 80+/-1, 107+/-1, and 144+/-5 mm Hg, respectively. In control rats, UNaV averaged 0.8+/ 0.4, 3.4+/-1.2, and 8.1+/-1.7 micromol/min at RPP of 77+/-2, 115+/-5, and 137+/-3 mm Hg, respectively. For normal and high RPP, UNaV was significantly lower in A 192621-treated rats compared with control rats. Additional experiments determined the effects of Big ET-1 (10 pmol/kg per minute) on intrarenal blood flow. Medullary blood flow (MBF) and cortical blood flow were measured in anesthetized rats by single-fiber, laser Doppler flowmetry. Cortical blood flow significantly decreased in response to Big ET-1 in rats on a normal or high salt diet. Big ET-1 significantly increased MBF in rats on a high salt diet, whereas there was no change in MBF in rats on a normal salt diet. These results demonstrate that medullary vasodilation produced by Big ET-1 is more prominent in rats on a high salt diet and are consistent with a contribution of ETB-mediated events in the natriuretic response to high salt intake. Taken together, these findings support the hypothesis that endothelin plays an important role in regulating sodium excretion through activation of ETB receptors. PMID- 12719444 TI - Decreases in ANP secretion by lysophosphatidylcholine through protein kinase C. AB - Lysophosphatidylcholine (LPC) is an endogenous phospholipid released from the cell membrane during ischemia, and it has potent, local effects on cardiac tissues. LPC has been implicated in arrhythmogenesis during ischemia by increasing intracellular Ca2+. However, it is not known whether LPC influences atrial release of atrial natriuretic peptide (ANP). The aim of this study was to investigate the effect of LPC on ANP secretion from isolated, perfused, beating rat atria. LPC (10 and 30 micromol/L) caused decreases in ANP secretion in a dose dependent manner, with slight increases in intra-atrial pressure and extracellular fluid (ECF) translocation. Therefore, the ANP secretion in terms of ECF translocation was markedly decreased by LPC. The order of the suppressive effect of ANP release was stearoyl-LPC>LPC>myristoyl-LPC=lauroyl-LPC. Staurosporine and wortmannin significantly attenuated suppression of the ANP release and an increase in intra-atrial pressure by LPC. High extracellular Mg2+ also attenuated the LPC-induced suppression of ANP release. However, other protein kinase C inhibitors such as chelerythrine, GF 109203X, and tamoxifen citrate did not affect LPC-induced suppression of ANP release. In single atrial myocytes, LPC caused increases in intracellular Ca2+ in a dose-dependent manner. The order of an increase in intracellular Ca2+ by LPC was stearoyl LPC>LPC>myristoyl-LPC=lauroyl-LPC. An increase in intracellular Ca2+ by LPC was attenuated by staurosporine. These results suggest that LPC-induced suppression of ANP release through protein kinase C/Ca2+ and phosphoinositol-3-kinase might in part play an important role in the development of hypertension. PMID- 12719445 TI - Heritability of blood pressure and hemodynamics in African- and European-American youth. AB - Hypertension prevalence is much higher in African-Americans (AAs) than in European-Americans (EAs). It is unknown whether this difference is related to potential ethnic differences in the relative contribution of genes and environment to population variation in blood pressure and underlying hemodynamics. We studied 308 EA and 226 AA twin pairs, including monozygotic and dizygotic twins, of the same as well as the opposite sex (mean+/-SD age, 14.7+/ 3.1 years). Supine resting systolic blood pressure (SBP), diastolic blood pressure (DBP), pulse pressure, and heart rate (HR) were measured by a Dinamap instrument and hemodynamics (stroke volume, cardiac index, and total peripheral resistance [TPR] index) by impedance cardiography. Ethnic and sex effects on genetic and environmental contributions to resting blood pressure and hemodynamics were estimated by genetic model fitting. For most measures, the best fitting model showed no differences in heritability between AAs and EAs or between males and females, with heritabilities of 0.50 for cardiac index, of 0.64 for HR, and of SBP, pulse pressure, and stroke volume in between. Heritability of DBP was 0.45 in EAs and 0.58 in AAs with no effect of sex. For TPR index in EAs, 46% of the variance could be attributed to familial effects, but no significant distinction could be made between shared environmental and genetic factors. Heritability of TPR index in AAs was 0.51. Adjustment for obesity yielded virtually identical heritabilities. In summary, relative influences of genetic and environmental factors on blood pressure and hemodynamics in AA and EA youth are similar and independent of (genes for) obesity. PMID- 12719446 TI - Recreational physical activity during pregnancy and risk of preeclampsia. AB - The potential benefits and risks of physical activity before and during pregnancy are not well studied. We studied the relation between recreational physical activity and the risk of preeclampsia in a case-control study of 201 preeclamptic and 383 normotensive pregnant women. Participants provided information about the type, intensity, frequency, and duration of physical activity performed during the first 20 weeks of pregnancy and during the year before pregnancy. Women who engaged in any regular physical activity during early pregnancy, compared with inactive women, experienced a 35% reduced risk of preeclampsia (odds ratio, 0.65; 95% confidence interval [CI], 0.43 to 0.99). Compared with inactive women, those engaged in light or moderate activities (ie, activities with metabolic-equivalent scores <6) experienced a 24% reduced risk of preeclampsia (95% CI, 0.48 to 1.20). The corresponding reduction for women participating in vigorous activities (metabolic equivalent scores > or =6) was 54% (95% CI, 0.27 to 0.79). Brisk walking (average walking pace > or =3 mi/h), when compared with no walking at all, was associated with a 30% to 33% reduction in preeclampsia risk. Stair climbing was inversely associated with the risk of preeclampsia (P for trend=0.039). Recreational physical activity performed during the year before pregnancy was associated with similar reductions in preeclampsia risk. These data suggest that regular physical activity, particularly when performed during the year before pregnancy and during early pregnancy, is associated with a reduced risk of preeclampsia. PMID- 12719447 TI - Selective involvement of p130Cas/Crk/Pyk2/c-Src in endothelin-1-induced JNK activation. AB - Both integrin-based focal adhesion complexes and receptor tyrosine kinases have been proposed as scaffolds on which the G protein-coupled receptor (GPCR)-induced signaling complex might assemble. We have recently reported that Ca2+-sensitive tyrosine kinase, Pyk2, and epidermal growth factor receptor (EGFR) act as independently regulated scaffolds in cardiomyocytes. In this report, we investigated the activation and regulation of p130Cas, Crk, Pyk2, and c-Src by a well-known hypertrophic agonist, endothelin-1 (ET), and determined their contributions to the activation of c-Jun NH2-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) in cardiomyocytes. Like Pyk2, ET induced tyrosine phosphorylation of p130Cas was significantly inhibited by either chelating intracellular Ca2+ ([Ca2+]i) or a protein kinase C inhibitor, calphostin C. This activation of p130Cas was also abrogated by the tetrapeptide RGDS, which disrupts integrin heterodimerization; cytochalasin D, which depolymerizes the actin cytoskeleton; or a selective Src family kinase inhibitor, PP2, but not by an EGFR inhibitor, AG1478. We also observed ET-induced temporal associations of Pyk2 with active c-Src, followed by p130Cas with Pyk2, c-Src, and Crk. Overexpression of a dominant-negative mutant of p130Cas (CasDeltaSD), Crk (CrkSH2m), Pyk2 (PKM), or C-terminal Src kinase (Csk), but not of a deletion mutant of EGFR (533delEGFR), attenuated ET-induced JNK activation. Similarly, an ET-induced increase in c-jun promoter luciferase activity was inhibited by overexpression of CasDeltaSD, CrkSH2m, PKM, or Csk. In contrast, ET-induced ERK activation and c-fos gene expression were predominantly regulated by EGFR. Collectively, the focal adhesion-dependent p130Cas/Crk/Pyk2/c-Src-mediated pathway is selectively involved in ET-induced JNK activation in cardiomyocytes. PMID- 12719448 TI - Endoscopic biopsies. PMID- 12719449 TI - Sidney Dyke. PMID- 12719451 TI - Guidelines for processing and reporting of prostatic needle biopsies. AB - The reported detection rate of prostate cancer, lesions suspicious for cancer, and prostatic intraepithelial neoplasia (PIN) in needle biopsies is highly variable. In part, technical factors, including the quality of the biopsies, the tissue processing, and histopathological reporting, may account for these differences. It has been thought that standardisation of tissue processing might reduce the observed variations in detection rate. Consensus among the members of the pathology committee of the European Randomised study of Screening for Prostate Cancer (ERSPC) concerning the optimal methodology of tissue embedding resulting in guidelines for prostatic needle biopsy processing was reached. The adoption of an unequivocal and uniform way of reporting lesions encountered in prostatic needle biopsies is considered helpful for decision taking by the clinician. The definition of parameters for quality control of prostatic needle biopsy diagnostics will further facilitate clinical epidemiological multicentre studies of prostate cancer. PMID- 12719450 TI - Nodal staging of colorectal carcinomas and sentinel nodes. AB - This review surveys the staging systems used for the classification of colorectal carcinomas, including the TNM system, and focuses on the assessment of the nodal stage of the disease. It reviews the quantitative requirements for a regional metastatic work up, and some qualitative features of lymph nodes that may help in the selection of positive and negative lymph nodes. Identification of the sentinel lymph nodes (those lymph nodes that have direct drainage from the primary tumour site) is one such qualitative feature that is claimed to allow the upstaging of colorectal carcinomas via an oriented, enhanced pathological work up. Current evidence in favour of a change in the requisite of assessing as may lymph nodes as is possible, and concentrating the efforts on only a selected number of lymph nodes, is weak. PMID- 12719452 TI - Variations in the processing of prostatic needle cores in the UK; what is safe? AB - AIMS: To determine the variation in the processing of prostatic needle cores in the UK and to compare the results with suggested guidelines. METHODS: A standard questionnaire was sent to 210 pathology departments enquiring about current practices. RESULTS: One hundred and thirty replies were received, which showed considerable variation in current methods. The number of cores received for each case ranged from three to 21, with the number of cores processed for each cassette varying from one to 10. Sixty per cent of centres used no special embedding techniques, and the number of sections cut for each case varied from two to 128, with a median of 12 sections for each case. Forty two per cent of laboratories did not take spare slides for immunochemistry. CONCLUSIONS: There is great variation in the processing of prostatic cores in the UK. In particular, some laboratories process a large number of cores in each cassette and do not use special embedding techniques. At present there are no established guidelines for the processing of these specimens. Enhanced techniques may well increase the sensitivity of the test but would increase the workload and costs to the pathology department. In view of the increasing workload from these specimens, a consensus for their optimum processing is required. PMID- 12719453 TI - National surveillance programme on susceptibility patterns of respiratory pathogens in South Africa: moxifloxacin compared with eight other antimicrobial agents. AB - AIMS: The susceptibility patterns of Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Klebsiella pneumoniae, and Streptococcus pyogenes isolated from specimens submitted to 12 private laboratories in South Africa were determined. METHODS: Minimum inhibitory concentration (MIC) determinations were performed on the isolates in the microbiology laboratory at Tygerberg Hospital according to the recommendations of the National Committee for Clinical Laboratory Standards (NCCLS). RESULTS: According to the NCCLS breakpoints, 24% of 729 S pneumoniae isolates were sensitive, 30% intermediate, and 46% resistant to penicillin. Rates of macrolide resistance were high, with 61% of the pneumococci being resistant to clarithromycin and azithromycin. Co trimoxazole resistance was also high, with 28% of pneumococcal strains being sensitive, 21% intermediate, and 51% resistant. beta Lactamase was produced by 7% of 736 H influenzae isolates and 91% of 256 M catarrhalis isolates. The quinolones, moxifloxacin and levofloxacin, were universally active against all isolates tested, which included S pneumoniae, H influenzae, M catarrhalis, K pneumoniae, and S pyogenes. CONCLUSIONS: Haemophilus influenzae and S pneumoniae were the most commonly isolated organisms. Resistance to penicillin was one of the highest reported in the world (76%) in S pneumoniae, as was macrolide resistance in pneumonocci, although surprisingly, only 14% of S pyogenes were resistant. The quinolones, moxifloxacin and levofloxacin, were active against all organisms tested, including the penicillin and macrolide resistant strains and moxifloxacin was more active than levofloxacin against pneumococci. PMID- 12719455 TI - Defensin pattern in chronic gastritis: HBD-2 is differentially expressed with respect to Helicobacter pylori status. AB - BACKGROUND/AIMS: Recent reports have suggested that Helicobacter pylori infection induces the mucosal antibiotic peptide human beta defensin 2 (HBD-2). Therefore, the present study investigated mRNA and peptide expression of four different defensins in the upper gastrointestinal tract in patients with H pylori positive and negative chronic gastritis. MATERIALS/METHODS: Biopsies from the oesophagus to the duodenum were taken during routine gastroscopy in 71 individuals. Total RNA was extracted and the reverse transcription polymerase chain reaction was performed with primers for human defensins 5 and 6 (HD-5/6) or HBD-1 and HBD-2. Paraffin wax embedded tissue from gastric resections was tested for HD-5, HBD-1, and HBD-2 by immunohistochemistry. RESULTS: Helicobacter pylori colonisation was associated with an increased percentage of positive biopsies with respect to HBD 2 in the corpus (p < 0.05). Helicobacter pylori had no impact on the gastric expression of HD-5 and HBD-1, whereas HD-6 was increased in the fundus. The abundant expression of alpha defensins in the duodenum and beta defensins in the oesophagus served as a positive control in each individual. Immunohistochemical analysis confirmed the presence of the HD-5, HBD-1, and HBD-2 peptides in gastric resection specimens. CONCLUSIONS: The recently described induction of HBD-2 upon H pylori infection was confirmed in a clinical setting of chronic gastritis. This phenomenon may be mediated by components of the pathogen itself or may occur secondary to immune events in chronic inflammation. PMID- 12719454 TI - Helicobacter pylori water soluble surface proteins prime human neutrophils for enhanced production of reactive oxygen species and stimulate chemokine production. AB - BACKGROUNDS/AIMS: Chronic gastritis induced by Helicobacter pylori is characterised by considerable neutrophil infiltration into the gastric mucosa without mucosal invasion of bacteria. Bacteria have different characteristics with respect to their ability to stimulate human neutrophils to produce reactive oxygen species and chemokines. The aim of this study was to examine the effects of H pylori water extracts on the oxidative burst and chemokine production of human neutrophils. METHODS: Helicobacter pylori cells were extracted by harvesting into distilled water and centrifugation. Neutrophils were incubated with H pylori water extracts and the production of reactive oxygen species was measured using luminol dependent chemiluminescence (LmCL). In addition, the concentrations of chemokines (interleukin 8 (IL-8), macrophage inflammatory protein 1-alpha (MIP1-alpha), and MIP1-beta) were measured by enzyme linked immunosorbent assay. Neutrophils were also stimulated by opsonised zymosan (OZ) after preincubation with H pylori water extracts. RESULTS: Helicobacter pylori water extracts alone induced only a weak oxidative burst but preincubation of neutrophils with water extracts dose dependently enhanced the LmCL response stimulated by OZ. Helicobacter pylori water extracts also stimulated neutrophil IL-8 production, although MIP-1beta production was only stimulated weakly, and MIP-1alpha was not stimulated at all. CONCLUSIONS: Helicobacter pylori products in water extracts may have a role in the activation and migration of neutrophils, which results in enhanced oxidative damage to gastric mucosa. These findings may explain the pathology of H pylori induced gastritis, in which there is little invasion of bacteria into the gastric mucosa. PMID- 12719456 TI - Increased apoptosis in gastric mucosa adjacent to intestinal metaplasia. AB - BACKGROUND: The biological processes involved in the development of gastric mucosal atrophy and intestinal metaplasia are still incompletely understood. Reports testing the hypothesis that apoptosis leads to atrophy have yielded conflicting results. The availability of new antibodies for the detection of apoptotic cells in tissue sections has facilitated the analysis of the role of apoptosis in the gastritis-atrophy-intestinal metaplasia sequence. METHODS: Archival material from 40 gastric resection specimens with normal mucosa (n = 5), chronic active gastritis (n = 17), or intestinal metaplasia (n = 18) was studied. Immunohistochemistry was performed using antibodies directed against cleaved cytokeratin 18 and active caspase 3. Slides were scored on a 0-3 scale for the presence of apoptotic cells. RESULTS: Normal gastric mucosa contained low numbers of apoptotic cells at the surface epithelium (mean score, 0.20). This number was significantly increased in cases with chronic gastritis (mean score, 1.06) and in those with intestinal metaplasia (mean score, 2.56). Within the intestinal metaplasia cases, 44 different foci of intestinal metaplasia were identified. In 39 of these 44 areas, concentrations of apoptotic cells were seen immediately adjacent to the foci of intestinal metaplasia, but not in the metaplastic epithelium itself. CONCLUSIONS: Apoptosis is uncommon in normal gastric mucosa. Chronic inflammation and intestinal metaplasia are associated with increased apoptosis, but occur mainly at the mucosal surface and not in the deeper layers. These findings do not support the concept that apoptosis underlies the loss of gastric glands and leads to atrophy, but the observed concentration of apoptotic epithelial cells adjacent to foci of intestinal metaplasia could be related to heterogeneity of epithelial damage, causing apoptosis, to which intestinal metaplasia is a response. PMID- 12719457 TI - Frequent occurrence of low grade cases among metastatic gastrointestinal stromal tumours. AB - AIM: Gastrointestinal stromal tumours (GISTs) are uncommon mesenchymal neoplasms. Some metastasise, whereas others remain asymptomatic for years, but it is difficult to distinguish between them histologically. This report analyses the characteristics of seven metastasising GISTs and compares clinicopathological parameters of the metastatic and non-metastatic groups. METHODS/RESULTS: Histology revealed typical GIST features with spindle, epithelioid, or mixed appearance. All seven cases were positive for vimentin, five for neurone specific enolase, six for c-kit, four for S-100, three for PGP-9.5, three for CD-34 and synaptophysin, but all were negative for cytokeratin, neurofilament, chromogranin A, and desmin. Four showed a focal reaction for smooth muscle actin. Three of the tumours were GI, and two each were GII and GIII. The Ki-67 index varied from 4% to 44%, the three GI cases had 4%, 10%, and 16%. Tumours from the metastatic GIST group were significantly larger than those from the non-metastatic group. CONCLUSIONS: Three cases exhibited bland, GI histological features with moderate or low proliferative activity. Among the c-kit positive metastasising stromal tumours, some were low grade, with moderate or low mitotic and Ki-67 indices, emphasising the necessity to develop a reliable grading system for GIST to predict clinical behaviour, the importance of careful analysis of "benign looking" tumours, and the key role of c-kit status in identifying patients who could benefit from treatment with STI-571. Larger tumours had a higher chance of metastasising, and only the size of the primary tumour played a role in predicting metastatic potential. PMID- 12719458 TI - Association between histology grade, expression of HsMCM2, and cyclin A in human invasive breast carcinomas. AB - AIM: Increased proliferation of tumour cells has prognostic value in human invasive breast carcinomas (IBCs), and high histology grade and cyclin A expression, which may reflect high proliferation rate, are associated with poor prognosis. Expression of HsMCM2 is related to cell proliferation. This study evaluates the correlation between the expression of cyclins A, D1, D3, and E, Ki 67, proliferating cell nuclear antigen (PCNA), histology grade, and HsMCM2 expression, in addition to the independent prognostic value of HsMCM2 expression in human IBCs. METHODS: Immunohistochemistry to evaluate HsMCM2, Ki-67, and PCNA expression in tumours from 147 patients with IBC. RESULTS: Nuclear staining for HsMCM2 was seen in 10-30% of the tumour cells in 30 samples, in 30-70% in 40 samples, in > 70% in 44 samples, and in < 10% in 33 samples. One way ANOVA showed a significant association between expression of HsMCM2 and cyclin A, D3, E, histology grade, and Ki-67. A borderline correlation was seen between HsMCM2 and PCNA. In multivariate analysis, the only association was with cyclin A, in addition to a borderline association with histology grade. In a Cox regression hazards model, expression of HsMCM2 was associated with poor patient survival, although it lost its independent prognostic value when cyclin A expression was included. Ki-67 and PCNA expression were not associated with patient survival. CONCLUSION: Cyclin A expression is independently associated with HsMCM2 expression, histology grade, and Ki-67. HsMCM2 expression is associated with poor patient survival, although it loses prognostic value when adjusted for cyclin A. PMID- 12719459 TI - Evaluation of a protocol for examining nephrectomy specimens with renal cell carcinoma. AB - AIMS: To evaluate the practicality of use and the effectiveness of a standard protocol for examining nephrectomy specimens for renal cell carcinoma (RCC), with emphasis on the identification of vascular invasion. METHODS: A standard protocol, devised to identify the major prognostic determinants, was used to examine 79 consecutive tumours submitted to four histopathology departments. The incidence of vascular invasion found was compared with the incidence in a historical series of tumours. RESULTS: The protocol proved easy to follow, and appeared to increase the incidence of observed vascular invasion (40 of 69 cases compared with 69 of 176 cases in the historical series; p = 0.059, Fishers exact test, one sided) CONCLUSIONS: If pathological prognostic determinants are to be used for clinical management, then it is important that they are identified and recorded consistently. The protocol described provides a method of examining nephrectomy specimens that can be used in routine practice and would probably reliably identify recognised prognostic variables. PMID- 12719460 TI - Alterations of MUC1 and MUC3 expression in gastric carcinoma: relevance to patient clinicopathological features. AB - AIMS: Several studies have reported conflicting and inconclusive results concerning the clinical relevance of mucin expression in gastric carcinoma. This study investigated the correlations between aberrant expression of mucins in gastric carcinoma and patient clinicopathological features. METHODS: The expression of MUC1, MUC2, MUC3, MUC5AC, and MUC6 was investigated immunohistochemically in gastric carcinoma (n = 46) in relation to patient clinicopathological features. RESULTS: All normal gastric mucosa samples expressed MUC1, MUC5AC, and MUC6. MUC1, MUC2, MUC3, MUC5AC, and MUC6 were expressed in 29, 31, 30, 18, and 21 of the 46 cases of gastric carcinoma, respectively. The number of cases expressing MUC1 was significantly higher (p < 0.01) in patients with a small tumour size (>/= 5 cm) and in patients in clinical stages I-II, compared with clinical stages III-IV (p < 0.05). Expression was significantly lower (p < 0.05) in patients exhibiting metastasis. The number of cases expressing MUC3 was significantly higher in patients in clinical stages III IV (p < 0.05), and in those with serosal invasion (p < 0.05) or metastasis (p<0.01). No significant relations were found between MUC2, MUC5AC, MUC6, and clinical stage, metastasis, or tumour size. CONCLUSIONS: Membrane bound mucins MUC1 and MUC3 appear to be associated with the development of gastric carcinoma. Patients who maintained high immunoreactivity for anti-MUC1 antibody had a better prognosis, whereas those with an increase in anti-MUC3 immunoreactivity had a poorer prognosis, as judged by tumour size, serosal invasion, and metastasis. However, no correlation was found between MUC2, MUC5AC, or MUC6 and clinical prognosis. PMID- 12719461 TI - Seasonal pseudohyperkalaemia. AB - AIMS: To investigate the relation between ambient temperature and serum potassium concentrations in samples from primary care. METHODS: Potassium concentrations were estimated on general practitioner and hospital ward samples taken over a two year period using serum obtained from gel separator samples. The number of samples analysed from general practice during each month varied from 5093 to 8978 (mean of 7068 samples/month). RESULTS: As the temperature fell in winter, the mean daily serum potassium concentration rose in samples from general practice, with the inverse occurring during the warmer summer months. This effect was restricted to samples coming from general practice, with inpatient samples seemingly unaffected. CONCLUSIONS: These results indicate that exposure of the samples to variations in ambient temperature during their transport to the laboratory profoundly affects measured serum potassium concentrations. PMID- 12719463 TI - Artifactually low glycated haemoglobin in a patient with severe hypertriglyceridaemia. AB - This report describes a case of artifactually low glycated haemoglobin (Hb) in a patient with type II diabetes and severe hypertriglyceridaemia. The effect of hypertriglyceridaemia on glycated Hb determination using the Abbott Vision method was investigated in a series of patients with diabetes. The interference of triglycerides in glycated Hb assays was also investigated by two other methods, the Beckman Synchron CX4 delta immunoturbidimetric method, and the Primus affinity chromatography high performance liquid chromatography assay. PMID- 12719462 TI - The role of antitissue transglutaminase assay for the diagnosis and monitoring of coeliac disease: a French-Italian multicentre study. AB - AIMS: Tissue transglutaminase (tTG) was recently identified as the major autoantigen in coeliac disease. The aim of this multicentre study was to evaluate the impact of a new immunoenzymatic assay for the detection of IgA anti-tGT antibodies. METHODS: Seventy four Italian and French clinical laboratories participated in this study; anti-tTG IgA with an enzyme linked immunosorbent assay (ELISA) method using guinea pig liver extract as the coating antigen, anti endomysium IgA autoantibodies (EMA), and total serum IgA were determined in 7948 patients, 1162 of whom had coeliac disease (737 untreated cases and 425 on a gluten free diet). A proportion of the sera were then sent to a reference laboratory for anti-tTG retesting with an ELISA method using recombinant human tTG antigen. RESULTS: Seven thousand four hundred and fifty eight (93.8%) sera were EMA/antiguinea pig tTG concordant (positive or negative); 490 (6.2%) were non-concordant. The sensitivity of EMA and antiguinea pig tTG in the 737 untreated patients with coeliac disease was 92.1% and 94.8%, respectively, and the specificity was 99.8% and 99.2%, respectively. Retesting of the discordant sera showed that of the 162 sera classified as EMA negative/antiguinea pig tTG positive, only 49 were positive for human recombinant anti-tTG, and that 39 of these were also EMA positive. Furthermore, of the 36 sera classified as EMA positive/antiguinea pig tTG negative, only two were confirmed as EMA positive. CONCLUSIONS: The antiguinea pig tTG assay is more sensitive but less specific than EMA, whereas the antihuman recombinant tTG assay is far more specific and just as sensitive as antiguinea pig tTG. Testing for EMA presents considerable interpretative problems and is difficult to standardise. PMID- 12719464 TI - A case of solitary pulmonary lymphangioma. AB - Solitary pulmonary lymphangiomas are rare benign lesions thought to result from the development of abnormally proliferating lymphatic vessels. This report describes a case of solitary pulmonary lymphangioma resected under video assisted thoracoscopic surgery and diagnosed using histological and immunohistochemical investigations. PMID- 12719465 TI - Internet referencing. PMID- 12719466 TI - Molten gold was poured down his throat until his bowels burst. PMID- 12719467 TI - Active repression by unliganded retinoid receptors in development: less is sometimes more. AB - The retinoid receptors have major roles throughout development, even in the absence of ligand. Here, we summarize an emerging theme whereby gene repression, mediated by unliganded retinoid receptors, can dictate cell fate. In addition to activating transcription, retinoid receptors actively repress gene transcription by recruiting cofactors that promote chromatin compaction. Two developmental processes for which gene silencing by the retinoid receptors is essential are head formation in Xenopus and skeletal development in the mouse. Inappropriate repression, by oncogenic retinoic acid (RA)**Abbreviations used in this paper: APL, acute promyelocytic leukemia; dnRARalpha, dominant-negative version of the RARalpha; E, embryonic age; HDAC, histone deacetylase; LCoR, ligand-dependent corepressor; NCoR, nuclear receptor corepressor; RA, retinoic acid; RAR, RA receptor; RARE, RXR homodimer bound to bipartite response element; RXR, retinoid X receptor; TSA, trichostatin A; CYP26, cytochrome p450, 26; TR, thyroid hormone receptor. receptor (RAR) fusion proteins, blocks myeloid differentiation leading to a rare form of leukemia. Our current understanding of the developmental role of retinoid repression and future perspectives in this field are discussed. PMID- 12719468 TI - Different degrees of lever arm rotation control myosin step size. AB - Myosins are actin-based motors that are generally believed to move by amplifying small structural changes in the core motor domain via a lever arm rotation of the light chain binding domain. However, the lack of a quantitative agreement between observed step sizes and the length of the proposed lever arms from different myosins challenges this view. We analyzed the step size of rat myosin 1d (Myo1d) and surprisingly found that this myosin takes unexpectedly large steps in comparison to other myosins. Engineering the length of the light chain binding domain of rat Myo1d resulted in a linear increase of step size in relation to the putative lever arm length, indicative of a lever arm rotation of the light chain binding domain. The extrapolated pivoting point resided in the same region of the rat Myo1d head domain as in conventional myosins. Therefore, rat Myo1d achieves its larger working stroke by a large calculated approximately 90 degrees rotation of the light chain binding domain. These results demonstrate that differences in myosin step sizes are not only controlled by lever arm length, but also by substantial differences in the degree of lever arm rotation. PMID- 12719469 TI - Helicobacter pylori CagA protein targets the c-Met receptor and enhances the motogenic response. AB - Infection with the human microbial pathogen Helicobacter pylori is assumed to lead to invasive gastric cancer. We find that H. pylori activates the hepatocyte growth factor/scatter factor receptor c-Met, which is involved in invasive growth of tumor cells. The H. pylori effector protein CagA intracellularly targets the c Met receptor and promotes cellular processes leading to a forceful motogenic response. CagA could represent a bacterial adaptor protein that associates with phospholipase Cgamma but not Grb2-associated binder 1 or growth factor receptor bound protein 2. The H. pylori-induced motogenic response is suppressed and blocked by the inhibition of PLCgamma and of MAPK, respectively. Thus, upon translocation, CagA modulates cellular functions by deregulating c-Met receptor signaling. The activation of the motogenic response in H. pylori-infected epithelial cells suggests that CagA could be involved in tumor progression. PMID- 12719470 TI - Aurora B couples chromosome alignment with anaphase by targeting BubR1, Mad2, and Cenp-E to kinetochores. AB - The Aurora/Ipl1 family of protein kinases plays multiple roles in mitosis and cytokinesis. Here, we describe ZM447439, a novel selective Aurora kinase inhibitor. Cells treated with ZM447439 progress through interphase, enter mitosis normally, and assemble bipolar spindles. However, chromosome alignment, segregation, and cytokinesis all fail. Despite the presence of maloriented chromosomes, ZM447439-treated cells exit mitosis with normal kinetics, indicating that the spindle checkpoint is compromised. Indeed, ZM447439 prevents mitotic arrest after exposure to paclitaxel. RNA interference experiments suggest that these phenotypes are due to inhibition of Aurora B, not Aurora A or some other kinase. In the absence of Aurora B function, kinetochore localization of the spindle checkpoint components BubR1, Mad2, and Cenp-E is diminished. Furthermore, inhibition of Aurora B kinase activity prevents the rebinding of BubR1 to metaphase kinetochores after a reduction in centromeric tension. Aurora B kinase activity is also required for phosphorylation of BubR1 on entry into mitosis. Finally, we show that BubR1 is not only required for spindle checkpoint function, but is also required for chromosome alignment. Together, these results suggest that by targeting checkpoint proteins to kinetochores, Aurora B couples chromosome alignment with anaphase onset. PMID- 12719471 TI - Sim4: a novel fission yeast kinetochore protein required for centromeric silencing and chromosome segregation. AB - Fission yeast centromeres are composed of two domains: the central core and the outer repeats. Although both regions are required for full centromere function, the central core has a distinct chromatin structure and is likely to underlie the kinetochore itself, as it is associated with centromere-specific proteins. Genes placed within either region are transcriptionally silenced, reflecting the formation of a functional kinetochore complex and flanking centromeric heterochromatin. Here, transcriptional silencing was exploited to identify components involved in central core silencing and kinetochore assembly or structure. The resulting sim (silencing in the middle of the centromere) mutants display severe chromosome segregation defects. sim2+ encodes a known kinetochore protein, the centromere-specific histone H3 variant Cnp1CENP-A. sim4+ encodes a novel essential coiled-coil protein, which is specifically associated with the central core region and is required for the unusual chromatin structure of this region. Sim4 coimmunoprecipitates with the central core component Mis6 and, like Mis6, affects Cnp1CENP-A association with the central domain. Functional Mis6 is required for Sim4 localization at the kinetochore. Our analyses illustrate the fundamental link between silencing, chromatin structure, and kinetochore function, and establish defective silencing as a powerful approach for identifying proteins required to build a functional kinetochore. PMID- 12719472 TI - Distinct in vivo roles for double-stranded RNA-binding domains of the Xenopus RNA editing enzyme ADAR1 in chromosomal targeting. AB - The RNA-editing enzyme adenosine deaminase that acts on RNA (ADAR1) deaminates adenosines to inosines in double-stranded RNA substrates. Currently, it is not clear how the enzyme targets and discriminates different substrates in vivo. However, it has been shown that the deaminase domain plays an important role in distinguishing various adenosines within a given substrate RNA in vitro. Previously, we could show that Xenopus ADAR1 is associated with nascent transcripts on transcriptionally active lampbrush chromosomes, indicating that initial substrate binding and possibly editing itself occurs cotranscriptionally. Here, we demonstrate that chromosomal association depends solely on the three double-stranded RNA-binding domains (dsRBDs) found in the central part of ADAR1, but not on the Z-DNA-binding domain in the NH2 terminus nor the catalytic deaminase domain in the COOH terminus of the protein. Most importantly, we show that individual dsRBDs are capable of recognizing different chromosomal sites in an apparently specific manner. Thus, our results not only prove the requirement of dsRBDs for chromosomal targeting, but also show that individual dsRBDs have distinct in vivo localization capabilities that may be important for initial substrate recognition and subsequent editing specificity. PMID- 12719473 TI - LST8 negatively regulates amino acid biosynthesis as a component of the TOR pathway. AB - LST8, a Saccharomyces cerevisiae gene encoding a 34-kD WD-repeat protein, was identified by mutations that caused defects in sorting Gap1p to the plasma membrane. Here, we report that the Gap1p sorting defect in the lst8-1 mutant results from derepression of Rtg1/3p activity and the subsequent accumulation of high levels of intracellular amino acids, which signal Gap1p sorting to the vacuole. To identify the essential function of Lst8p, we isolated lst8 mutants that are temperature-sensitive for growth. These mutants show hypersensitivity to rapamycin and derepressed Gln3p activity like cells with compromised TOR pathway activity. Like tor2 mutants, lst8 mutants also have cell wall integrity defects. Confirming a role for Lst8p in the TOR pathway, we find that Lst8p associates with both Tor1p and Tor2p and is a peripheral membrane protein that localizes to endosomal or Golgi membranes and cofractionates with Tor1p. Further, we show that a sublethal concentration of rapamycin mimics the Gap1p sorting defect of an lst8 mutant. Finally, the different effects of lst8 alleles on the activation of either the Rtg1/3p or Gln3p transcription factors reveal that these two pathways constitute distinct, genetically separable outputs of the Tor-Lst8 regulatory complex. PMID- 12719474 TI - Identification of XMAP215 as a microtubule-destabilizing factor in Xenopus egg extract by biochemical purification. AB - Microtubules (MTs) polymerized with GMPCPP, a slowly hydrolyzable GTP analogue, are stable in buffer but are rapidly depolymerized in Xenopus egg extracts. This depolymerization is independent of three previously identified MT destabilizers (Op18, katanin, and XKCM1/KinI). We purified the factor responsible for this novel depolymerizing activity using biochemical fractionation and a visual activity assay and identified it as XMAP215, previously identified as a prominent MT growth-promoting protein in Xenopus extracts. Consistent with the purification results, we find that XMAP215 is necessary for GMPCPP-MT destabilization in extracts and that recombinant full-length XMAP215 as well as an NH2-terminal fragment have depolymerizing activity in vitro. Stimulation of depolymerization is specific for the MT plus end. These results provide evidence for a robust MT destabilizing activity intrinsic to this microtubule-associated protein and suggest that destabilization may be part of its essential biochemical functions. We propose that the substrate in our assay, GMPCPP-stabilized MTs, serves as a model for the pause state of MT ends and that the multiple activities of XMAP215 are unified by a mechanism of antagonizing MT pauses. PMID- 12719475 TI - Stu2p, the budding yeast member of the conserved Dis1/XMAP215 family of microtubule-associated proteins is a plus end-binding microtubule destabilizer. AB - The Dis1/XMAP215 family of microtubule-associated proteins conserved from yeast to mammals is essential for cell division. XMAP215, the Xenopus member of this family, has been shown to stabilize microtubules in vitro, but other members of this family have not been biochemically characterized. Here we investigate the properties of the Saccharomyces cerevisiae homologue Stu2p in vitro. Surprisingly, Stu2p is a microtubule destabilizer that binds preferentially to microtubule plus ends. Quantitative analysis of microtubule dynamics suggests that Stu2p induces microtubule catastrophes by sterically interfering with tubulin addition to microtubule ends. These results reveal both a new biochemical activity for a Dis1/XMAP215 family member and a novel mechanism for microtubule destabilization. PMID- 12719476 TI - Shear stress-induced endothelial cell polarization is mediated by Rho and Rac but not Cdc42 or PI 3-kinases. AB - Shear stress induces endothelial polarization and migration in the direction of flow accompanied by extensive remodeling of the actin cytoskeleton. The GTPases RhoA, Rac1, and Cdc42 are known to regulate cell shape changes through effects on the cytoskeleton and cell adhesion. We show here that all three GTPases become rapidly activated by shear stress, and that each is important for different aspects of the endothelial response. RhoA was activated within 5 min after stimulation with shear stress and led to cell rounding via Rho-kinase. Subsequently, the cells respread and elongated within the direction of shear stress as RhoA activity returned to baseline and Rac1 and Cdc42 reached peak activation. Cell elongation required Rac1 and Cdc42 but not phosphatidylinositide 3-kinases. Cdc42 and PI3Ks were not required to establish shear stress-induced polarity although they contributed to optimal migration speed. Instead, Rho and Rac1 regulated directionality of cell movement. Inhibition of Rho or Rho-kinase did not affect the cell speed but significantly increased cell displacement. Our results show that endothelial cells reorient in response to shear stress by a two step process involving Rho-induced depolarization, followed by Rho/Rac-mediated polarization and migration in the direction of flow. PMID- 12719477 TI - Expression of a targeted lambda 1 light chain gene is developmentally regulated and independent of Ig kappa rearrangements. AB - Immunoglobulin light chain (IgL) rearrangements occur more frequently at Ig kappa than at Ig lambda. Previous results suggested that the unrearranged Ig kappa locus negatively regulates Ig lambda transcription and/or rearrangement. Here, we demonstrate that expression of a VJ lambda 1-joint inserted into its physiological position in the Ig lambda locus is independent of Ig kappa rearrangements. Expression of the inserted VJ lambda 1 gene segment is developmentally controlled like that of a VJ kappa-joint inserted into the Ig kappa locus and furthermore coincides developmentally with the occurrence of Ig kappa rearrangements in wild-type mice. We conclude that developmentally controlled transcription of a gene rearrangement in the Ig lambda locus occurs in the presence of an unrearranged Ig kappa locus and is therefore not negatively regulated by the latter. Our data also indicate light chain editing in approximately 30% of lambda 1 expressing B cell progenitors. PMID- 12719478 TI - Dectin-1 mediates the biological effects of beta-glucans. AB - The ability of fungal-derived beta-glucan particles to induce leukocyte activation and the production of inflammatory mediators, such as tumor necrosis factor (TNF)-alpha, is a well characterized phenomenon. Although efforts have been made to understand how these carbohydrate polymers exert their immunomodulatory effects, the receptors involved in generating these responses are unknown. Here we show that Dectin-1 mediates the production of TNF-alpha in response to zymosan and live fungal pathogens, an activity that occurs at the cell surface and requires the cytoplasmic tail and immunoreceptor tyrosine activation motif of Dectin-1 as well as Toll-like receptor (TLR)-2 and Myd88. This is the first demonstration that the inflammatory response to pathogens requires recognition by a specific receptor in addition to the TLRs. Furthermore, these studies implicate Dectin-1 in the production of TNF-alpha in response to fungi, a critical step required for the successful control of these pathogens. PMID- 12719479 TI - Collaborative induction of inflammatory responses by dectin-1 and Toll-like receptor 2. AB - Toll-like receptors (TLRs) mediate recognition of a wide range of microbial products including lipopolysaccharides, lipoproteins, flagellin, and bacterial DNA, and signaling through TLRs leads to the production of inflammatory mediators. In addition to TLRs, many other surface receptors have been proposed to participate in innate immunity and microbial recognition, and signaling through some of these receptors is likely to cooperate with TLR signaling in defining inflammatory responses. In this report we have examined how dectin-1, a lectin family receptor for beta-glucans, collaborates with TLRs in recognizing microbes. Dectin-1, which is expressed at low levels on macrophages and high levels on dendritic cells, contains an immunoreceptor tyrosine-based activation motif-like signaling motif that is tyrosine phosphorylated upon activation. The receptor is recruited to phagosomes containing zymosan particles but not to phagosomes containing immunoglobulin G-opsonized particles. Dectin-1 expression enhances TLR-mediated activation of nuclear factor kappa B by beta-glucan containing particles, and in macrophages and dendritic cells dectin-1 and TLRs are synergistic in mediating production of cytokines such as interleukin 12 and tumor necrosis factor alpha. Additionally, dectin-1 triggers production of reactive oxygen species, an inflammatory response that is primed by TLR activation. The data demonstrate that collaborative recognition of distinct microbial components by different classes of innate immune receptors is crucial in orchestrating inflammatory responses. PMID- 12719480 TI - Molecular modeling and functional mapping of B7-H1 and B7-DC uncouple costimulatory function from PD-1 interaction. AB - B7-H1 and B7-DC are ligands for PD-1, a receptor implicated in negative regulation of T and B cell functions. These ligands, however, also costimulate T cell responses. It remains elusive whether or not costimulation is mediated through PD-1. By comparative molecular modeling and site-directed mutagenesis, we found that nonconserved residues between these ligands on the A'GFCC'C" face mediate interaction with PD-1. This indicates significant structural heterogeneity of the interactions between PD-1 and its ligands. Importantly, ligand mutants with abolished PD-1 binding capacity could still costimulate proliferation and cytokine production of T cells from normal and PD-1-deficient mice. Our results reveal unique binding characteristics of B7-H1 and B7-DC and provide direct evidence for an independent costimulatory receptor other than PD 1. PMID- 12719481 TI - Death by a B cell superantigen: In vivo VH-targeted apoptotic supraclonal B cell deletion by a Staphylococcal Toxin. AB - Amongst the many ploys used by microbial pathogens to interfere with host immune responses is the production of proteins with the properties of superantigens. These properties enable superantigens to interact with conserved variable region framework subdomains of the antigen receptors of lymphocytes rather than the complementarity determining region involved in the binding of conventional antigens. To understand how a B cell superantigen affects the host immune system, we infused protein A of Staphylococcus aureus (SpA) and followed the fate of peripheral B cells expressing B cell receptors (BCRs) with VH regions capable of binding SpA. Within hours, a sequence of events was initiated in SpA-binding splenic B cells, with rapid down-regulation of BCRs and coreceptors, CD19 and CD21, the induction of an activation phenotype, and limited rounds of proliferation. Apoptosis followed through a process heralded by the dissipation of mitochondrial membrane potential, the induction of the caspase pathway, and DNA fragmentation. After exposure, B cell apoptotic bodies were deposited in the spleen, lymph nodes, and Peyer's patches. Although in vivo apoptosis did not require the Fas death receptor, B cells were protected by interleukin (IL)-4 or CD40L, or overexpression of Bcl-2. These studies define a pathway for BCR mediated programmed cell death that is VH region targeted by a superantigen. PMID- 12719482 TI - Excitation contraction coupling in cardiac muscle: is there a purely voltage dependent component? PMID- 12719483 TI - The voltage-sensitive release mechanism of excitation contraction coupling in rabbit cardiac muscle is explained by calcium-induced calcium release. AB - The putative voltage-sensitive release mechanism (VSRM) was investigated in rabbit cardiac myocytes at 37 degrees C with high resistance microelectrodes to minimize intracellular dialysis. When the holding potential was adjusted from -40 to -60 mV, the putative VSRM was expected to operate alongside CICR. Under these conditions however, we did not observe a plateau at positive potentials of the cell shortening versus voltage relationship. The threshold for cell shortening changed by -10 mV, but this resulted from a similar change of the threshold for activation of inward current. Cell shortening under conditions where the putative VSRM was expected to operate was blocked in a dose dependent way by nifedipine and CdCl2 and blocked completely by NiCl2. "Tail contractions" persisted in the presence of nifedipine and CdCl2 but were blocked completely by NiCl2. Block of early outward current by 4-aminopyridine and 4-acetoamido-4'-isothiocyanato stilbene-2,2'-disulfonic acid (SITS) demonstrated persisting inward current during test depolarizations despite the presence of nifedipine and CdCl2. Inward current did not persist in the presence of NiCl2. A tonic component of cell shortening that was prominent during depolarizations to positive potentials under conditions selective for the putative VSRM was sensitive to rapidly applied changes in superfusate [Na+] and to the outward Na+/Ca2+ exchange current blocking drug KB-R7943. This component of cell shortening was thought to be the result of Na+/Ca2+ exchange-mediated excitation contraction coupling. Cell shortening recorded under conditions selective for the putative VSRM was increased by the enhanced state of phosphorylation induced by isoprenaline (1 microM) and by enhancing sarcoplasmic reticulum Ca2+ content by manipulation of the conditioning steps. Under these conditions, cell shortening at positive test depolarizations was converted from tonic to phasic. We conclude that the putative VSRM is explained by CICR with the Ca2+ "trigger" supplied by unblocked L-type Ca2+ channels and Na+/Ca2+ exchange. PMID- 12719485 TI - Activation and desensitization of the recombinant P2X1 receptor at nanomolar ATP concentrations. AB - Activation and desensitization kinetics of the rat P2X1 receptor at nanomolar ATP concentrations were studied in Xenopus oocytes using two-electrode voltage-clamp recording. The solution exchange system used allowed complete and reproducible solution exchange in <0.5 s. Sustained exposure to 1-100 nM ATP led to a profound desensitization of P2X1 receptors. At steady-state, desensitization could be described by the Hill equation with a K1/2 value of 3.2 +/- 0.1 nM. Also, the ATP dependence of peak currents could be described by a Hill equation with an EC50 value of 0.7 microM. Accordingly, ATP dose-effect relationships of activation and desensitization practically do not overlap. Recovery from desensitization could be described by a monoexponential function with the time-constant tau = 11.6 +/ 1.0 min. Current transients at 10-100 nM ATP, which elicited 0.1-8.5% of the maximum response, were compatible with a linear three-state model, C-O-D (closed open-desensitized), with an ATP concentration-dependent activation rate and an ATP concentration-independent (constant) desensitization rate. In the range of 18 300 nM ATP, the total areas under the elicited current transients were equal, suggesting that P2X1 receptor desensitization occurs exclusively via the open conformation. Hence, our results are compatible with a model, according to which P2X1 receptor activation and desensitization follow the same reaction pathway, i.e., without significant C to D transition. We assume that the K1/2 of 3.2 nM for receptor desensitization reflects the nanomolar ATP affinity of the receptor found by others in agonist binding experiments. The high EC50 value of 0.7 microM for receptor activation is a consequence of fast desensitization combined with nonsteady-state conditions during recording of peak currents, which are the basis of the dose-response curve. Our results imply that nanomolar extracellular ATP concentrations can obscure P2X1 receptor responses by driving a significant fraction of the receptor pool into a long-lasting refractory closed state. PMID- 12719484 TI - Inositol 1,4,5-trisphosphate-induced calcium release is necessary for generating the entire light response of limulus ventral photoreceptors. AB - The experiments reported here were designed to answer the question of whether inositol 1,4,5-trisphosphate (IP3)-induced calcium release is necessary for generating the entire light response of Limulus ventral photoreceptors. For this purpose the membrane-permeable IP3 receptor antagonist 2-aminoethoxydiphenyl borate (2APB) (Maruyama, T., T. Kanaji, S. Nakade, T. Kanno, and K. Mikoshiba. 1997. J. Biochem. (Tokyo). 122:498-505) was used. Previously, 2APB was found to inhibit the light activated current of Limulus ventral photoreceptors and reversibly inhibit both light and IP3 induced calcium release as well as the current activated by pressure injection of calcium into the light sensitive lobe of the photoreceptor (Wang, Y., M. Deshpande, and R. Payne. 2002. Cell Calcium. 32:209). In this study 2APB was found to inhibit the response to a flash of light at all light intensities and to inhibit the entire light response to a step of light, that is, both the initial transient and the steady-state components of the response to a step of light were inhibited. The light response in cells injected with the calcium buffer 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA) was reversibly inhibited by 2APB, indicating that these light responses result from IP3-mediated calcium release giving rise to an increase in Cai. The light response obtained from cells after treatment with 100 microM cyclopiazonic acid (CPA), which acts to empty intracellular calcium stores, was reversibly inhibited by 2APB, indicating that the light response after CPA treatment results from IP3-mediated calcium release and a consequent rise in Cai. Together these findings imply that IP3-induced calcium release is necessary for generating the entire light response of Limulus ventral photoreceptors. PMID- 12719488 TI - Medication errors in psychiatry: are patients being harmed? PMID- 12719486 TI - Tandem gramicidin channels cross-linked by streptavidin. AB - The interaction of biotin-binding proteins with biotinylated gramicidin (gA5XB) was studied by monitoring single-channel activity and sensitized photoinactivation kinetics. It was discovered that the addition of streptavidin or avidin to the bathing solutions of a bilayer lipid membrane (BLM) with incorporated gA5XB induced the opening of a channel characterized by approximately doubled single-channel conductance and extremely long open-state duration. We believe that the deceleration of the photoinactivation kinetics observed here with streptavidin and previously (Rokitskaya, T.I., Y.N. Antonenko, E.A. Kotova, A. Anastasiadis, and F. Separovic. 2000. Biochemistry. 39:13053 13058) with avidin reflects the formation of long-lived channels of this type. Both opening and closing of the double-conductance channels occurred via a transient sub-state of the conductance coinciding with that of the usual single channel transition. The appearance of the double-conductance channels after the addition of streptavidin was preceded by bursts of fast fluctuations of the current with the open state duration of the individual events of 60 ms. The streptavidin-induced double-conductance channels appeared to be inherent only to the gramicidin analogue with a biotin group linked to the COOH terminus through a long linker arm. Including biotinylated phosphatidylethanolamine into the BLM prevented the formation of the double-conductance channels even with the excess streptavidin. In view of the results obtained here, it is suggested that the double-conductance channel represents a tandem of two neighboring gA5XB channels with their COOH termini being cross-linked by the bound streptavidin at both sides of the BLM. The finding that streptavidin induces the formation of the tandem gramicidin channel comprising two channels functioning in concert is considered to be relevant to the physiologically important phenomenon of ligand induced receptor oligomerization. PMID- 12719490 TI - Datapoints: Mental health services in employee assistance programs, 2001. PMID- 12719491 TI - Alcohol & drug abuse: A research-based analysis of the Moderation Management controversy. PMID- 12719492 TI - Practical psychotherapy: Adaptation of dialectical behavior therapy by a VA Medical Center. PMID- 12719487 TI - Hydrophobic coupling of lipid bilayer energetics to channel function. AB - The hydrophobic coupling between membrane-spanning proteins and the lipid bilayer core causes the bilayer thickness to vary locally as proteins and other "defects" are embedded in the bilayer. These bilayer deformations incur an energetic cost that, in principle, could couple membrane proteins to each other, causing them to associate in the plane of the membrane and thereby coupling them functionally. We demonstrate the existence of such bilayer-mediated coupling at the single molecule level using single-barreled as well as double-barreled gramicidin channels in which two gramicidin subunits are covalently linked by a water soluble, flexible linker. When a covalently attached pair of gramicidin subunits associates with a second attached pair to form a double-barreled channel, the lifetime of both channels in the assembly increases from hundreds of milliseconds to a hundred seconds--and the conductance of each channel in the side-by-side pair is almost 10% higher than the conductance of the corresponding single barreled channels. The double-barreled channels are stabilized some 100,000-fold relative to their single-barreled counterparts. This stabilization arises from: first, the local increase in monomer concentration around a single-barreled channel formed by two covalently linked gramicidins, which increases the rate of double-barreled channel formation; and second, from the increased lifetime of the double-barreled channels. The latter result suggests that the two barrels of the construct associate laterally. The underlying cause for this lateral association most likely is the bilayer deformation energy associated with channel formation. More generally, the results suggest that the mechanical properties of the host bilayer may cause the kinetics of membrane protein conformational transitions to depend on the conformational states of the neighboring proteins. PMID- 12719493 TI - Rehab rounds: Involving families in rehabilitation through behavioral family management. AB - Introduction by the column editors: A wide variety of family psychoeducational approaches for persons with serious and persistent mental illness have been developed and used by mental health professionals since the introduction of such approaches in the 1960s (1). From 1970 to 1975, multifamily psychoeducation was implemented for large numbers of clients and relatives at the Oxnard Mental Health Center (2) and was followed by systematic studies of multifamily therapy at the Maudsley Hospital in London in 1976 (3) and Camarillo State Hospital in California in 1978 to 1980 (4). The type of intervention that arose from this work comprised a series of educational sessions followed by training in communication and problem-solving skills. Behavioral learning principles were used to promote acquisition of knowledge, coping skills, and problem solving for families. Other procedures and variants on this theme have proliferated throughout the United States and in many other countries (5). Most of these approaches have in common an adherence to practical goals that are individualized for each family and an educational rather than a "therapeutic" slant. In an attempt to overcome the obstacles to widespread clinical use of family interventions, a variety of dissemination strategies have been tried, including academic detailing, consensus building among all stakeholders (6), and the development of modules consisting of trainers' manuals, consumers' workbooks, slide shows, and video-assisted learning (7,8). This month's column describes one of the variants of family psychoeducation-behavioral family management-and its components, illuminated by case vignettes and results of efficacy studies. PMID- 12719494 TI - Best practices: Interpreting measurement data for quality improvement: standards, means, norms, and benchmarks. PMID- 12719495 TI - Psychopharmacology: Perspectives on medication adherence and atypical antipsychotic medications. PMID- 12719496 TI - Challenges for psychiatry in serving homeless people with psychiatric disorders. AB - The authors examine current challenges confronting psychiatry in caring for homeless people with psychiatric disorders. After reviewing how psychiatry has historically addressed homelessness and mental illness, the authors discuss the roles that the profession has developed in working with homeless populations. These roles, which encompass clinical, administrative, academic, and advocacy functions, have evolved as a result of trends both in homelessness services and within the profession of psychiatry. Challenges implicit in this evolution are discussed, including recent trends in homelessness, particularly an increase in prevalence, especially among families and children and some clinical subpopulations. The authors propose that these epidemiological trends are affecting the mental health care needs of homeless people. To be effective and credible in continuing to help solve the problems of homeless people with psychiatric disorders, psychiatry must adapt to these new challenges, using the roles it has developed. PMID- 12719497 TI - Use of chart and record reviews to detect medication errors in a state psychiatric hospital. AB - OBJECTIVE: This study compared the effectiveness of using a review team and the usual self-reporting method in detecting different types of medication errors in a state psychiatric hospital. METHODS: Medication errors were defined by using widely accepted criteria. Rates of prescription, transcription, administration, and dispensing errors were determined, and the risk of harm from each error was rated as high, moderate, or low. A review team was assigned to retrospectively review 31 patient records for prescription, transcription, and administration errors for a total of 1,448 patient-days. Dispensing errors, which can only be determined concurrently, were reported for an equivalent number of patient-days. The error rate was compared with the rate that was determined by the usual method of self-reports from all nursing and medical staff. RESULTS: In the 31 charts retrospectively reviewed and the dispensing events concurrently reviewed, the team detected a total of 2,194 medication errors, whereas a total of nine errors were self-reported for the same patient group. Administration errors accounted for more than half of the total (66 percent), followed by transcription errors (23 percent), prescription errors (11 percent), and dispensing errors (less than 1 percent). Nineteen percent of errors were rated as having a low risk of harm, 23 percent as having a moderate risk, and 58 percent as having a high risk. CONCLUSIONS: Use of a review team should be considered as a method for detecting and reporting medication errors. PMID- 12719498 TI - Aligning incentives in the treatment of depression in primary care with evidence based practice. AB - Deficits in the quality of treatment of depression in the primary care sector have been documented in multiple studies. Several clinical models for improving primary care treatment of depression have been shown to be cost-effective in recent years but have not proved to be sustainable over time, partly because of barriers created by common organizational and financing arrangements such as managed behavioral health care carve-outs and risk-based provider payment mechanisms. These arrangements, which often distort relative costs that primary care physicians face when making treatment decisions for patients who have depression, can steer these decisions away from evidence-based practice. Various changes, such as in contractual relationships, payment methods for primary care physicians, and performance measurement, can be made in existing institutional arrangements to better align them with emerging clinical technologies and evidence-based practice. PMID- 12719499 TI - The integral role of pastoral counseling by African-American clergy in community mental health. AB - OBJECTIVE: Little is currently known about the pastoral counseling work of pastors of African-American churches. The authors interviewed the pastors of nearly all African-American churches in a metropolitan area about their pastoral counseling work and related aspects of their ministry. METHODS: Of 121 African American pastors identified, 99 completed a semistructured interview describing their backgrounds, attitudes, concerns, and work. The interview included detailed queries about how they understood and carried out any pastoral counseling work. RESULTS: The respondents averaged more than six hours of counseling work weekly and often addressed serious problems similar to those seen by secular mental health professionals, with whom they reported readily exchanging referrals. Many of the respondents reported having and maintaining specialized education for their counseling work, which they described as including both spiritual and psychological dimensions. Most of the pastors reported that they observe and address severe mental illness and substance abuse in their congregations and that they also counsel individuals outside their own denominations. CONCLUSIONS: African-American urban ministers functioning as pastoral counselors constitute an engaging and useful group with experiences and skills that can be tapped by interested secular professionals. Their work represents a significant mental health resource for persons who lack sufficient access to needed care. PMID- 12719500 TI - A national study of the effect of chronic pain on the use of health care by depressed persons. AB - OBJECTIVE: The objective of this study was to provide national estimates of the impact of common pain conditions such as back pain, chronic headache, self reported arthritis, and unspecified chronic pain on the use of health services and quality of care among persons with depression. METHODS: This was a cross sectional study of data from a U.S. national household survey conducted in 1997 1998. The participants were 1,486 community-dwelling adults who met criteria for major depression or dysthymia according to the Composite International Diagnostic Interview Short-Form. RESULTS: Depressed persons with comorbid pain (N=938) were older, had lower incomes, and reported more severe psychiatric distress than depressed persons who did not have pain. When sociodemographic characteristics and severity of psychological distress were adjusted for, comorbid pain was associated with about 20 percent more visits to medical providers by patients who made at least one visit during a year. However, the patients with comorbid pain were about 20 percent less likely to see a mental health specialist than patients without pain. Pain was also shown to be associated with greater use of complementary or alternative medicine but not with differences in the use of antidepressants. CONCLUSION: Comorbid pain among persons with depression is associated with more intensive use of general medical services but lower rates of use of mental health services. PMID- 12719501 TI - Cost-effectiveness of collaborative care for depression in a primary care veteran population. AB - OBJECTIVE: This study examined the incremental cost-effectiveness of a collaborative care intervention for depression compared with consult-liaison care. METHODS: A total of 354 patients in a Department of Veterans Affairs (VA) primary care clinic who met the criteria for major depression or dysthymia were randomly assigned to one of the two care models. Under the collaborative care model, a mental health team provided a treatment plan to primary care providers, telephoned patients to encourage adherence, reviewed treatment results, and suggested modifications. Outcomes were assessed at three and nine months by telephone interviews. Health care use and costs were also assessed. RESULTS: A significantly greater number of collaborative care patients were treated for depression and given prescriptions for antidepressants. The collaborative care patients experienced an average of 14.6 additional depression-free days over the nine months. The mean incremental cost of the intervention per patient was $237 US dollars for depression treatment and $519 US dollars for total outpatient costs. A majority of the additional expenditures were accounted for by the intervention. The incremental cost-effectiveness ratio was $24 US dollars per depression-free day for depression treatment costs and $33 US dollars for total outpatient cost. CONCLUSIONS: Better coordination and communication under collaborative care was associated with a greater number of patients being treated for depression and with moderate increases in days free of depression and in treatment cost. Additional resources are needed for effective collaborative care models for depression treatment in primary care. PMID- 12719502 TI - Administrative burden and its implications for outpatient substance abuse treatment organizations. AB - OBJECTIVE: The aim of this study was to examine the extent of administrative burden on outpatient substance abuse treatment organizations and its implications for efficiency and productivity. METHODS: Using data from the 1995 and 2000 waves of the National Drug Abuse Treatment System Survey, the authors conducted multivariate analyses using generalized estimating equations. Two measures of organizational efficiency (operating expenses per therapy hour and salary and wages per therapy hour) and one measure of productivity (treatment sessions per full-time equivalent) were included. RESULTS: The average administrative burden in outpatient substance abuse treatment units increased between 1995 and 2000. The weighted and adjusted national sample data showed that one hour of substance abuse treatment therapy was associated with approximately $60 US dollars (in 1999 dollars) of nonsalary operating expenses and $124 US dollars in salaries and wages. Approximately eight treatment sessions were delivered each week by each full-time-equivalent staff member. The average weekly administrative burden consisted of about 71 hours of administrative work and almost 87 hours of clerical work. After controlling for various organizational characteristics, the authors found support for their predictions that administrative burden has negative relationships with organizational efficiency and productivity. CONCLUSIONS: This study uncovered different relationships for various types of burden and different types of organizational performance. It provided some evidence that, as treatment organizations face increased administrative burden, they may be shifting resources away from the provision of care. PMID- 12719503 TI - Impact of structured clinical interviews on physicians' practices in community mental health settings. AB - OBJECTIVE: Budgetary constraints often limit practicing psychiatrists from conducting more thorough diagnostic evaluations. This study examined physicians' diagnosing and prescribing practices in the context of feedback from nurse administered, structured clinical interviews of psychiatric patients in publicly funded outpatient mental health settings. METHODS: A randomized controlled trial was conducted of newly enrolled adult psychiatric patients who made at least one return visit for care at two county-supported outpatient clinics. Within two weeks after their intake psychiatric evaluation, patients were randomly assigned to receive a nurse-administered Structured Clinical Interview for DSM-IV Clinician Version (SCID) (N=158) or to a control condition (N=138). The attending psychiatrist was provided with the SCID results. Abstracts from clinical records were used to measure differences in physicians' rates of ordering additional diagnostic evaluations, changing diagnoses, and changing types and dosages of medications at three- and six-month follow-ups. RESULTS: Physicians treating patients who received SCIDs, compared with control patients, were more likely to order evaluative procedures, update and change diagnosis (consistent with SCID results), and change prescription medication type and were less likely to increase patients' medication dosages. CONCLUSION: Nurse-administered structured clinical interviews are feasible and effectively help psychiatrists in publicly supported mental health clinics reach more accurate diagnoses for newly enrolled patients. PMID- 12719504 TI - Treatment adherence associated with conventional and atypical antipsychotics in a large state Medicaid program. AB - OBJECTIVES: Rates of medication adherence over a one-year period were assessed among outpatients with schizophrenia who initiated therapy with conventional or atypical antipsychotic agents. METHODS: Data were drawn from paid medical and pharmacy claims for a random sample of 10 percent of all California Medicaid ("Medi-Cal") recipients. Outpatients with schizophrenia who were aged 18 years or older and who initiated monotherapy with a conventional or atypical antipsychotic medication in the last quarter of 1997 were identified. The percentages of patients who discontinued antipsychotic therapy or who had a switch in medications over a one-year period were determined. The use of selected concomitant medications was also assessed. Data were analyzed by means of chi square tests and multivariate statistics that adjusted for demographic and clinical differences between medication groups. RESULTS: A total of 298 patients who initiated therapy with a conventional (N=93) or atypical (N=205) antipsychotic medication were identified. The groups were similar in mean age (about 42 years) and gender distribution (about 54 percent were male). Compared with patients who received conventional antipsychotics, those receiving atypical antipsychotics were significantly less likely to have a switch in medication and to use concomitant anticholinergic and anxiolytic medications. In each group, antipsychotic medication was available for about 60 percent of days over one year of follow-up. CONCLUSIONS: Compared with the use of conventional antipsychotics, the use of atypical antipsychotic medications was associated with significantly less treatment switching and less use of concomitant medications. However, undertreatment, evidenced by a lack of prescription refills, occurred among patients taking both medication classes, which highlights the need for further research on nonadherence. PMID- 12719505 TI - Use of antidepressants among Canadian workers receiving depression-related short term disability benefits. AB - OBJECTIVES: Little is known about how antidepressants are being used, but rising antidepressant expenditures and the accompanying impulse to control costs make this a critical issue to be addressed. The authors studied patterns of antidepressant use in a population of workers receiving depression-related short term disability benefits to determine whether populations likely to benefit from antidepressants are using them and, if so, whether they are using them in a way that the benefits from their use are maximized. METHODS: The analyses were based on 1996-1998 administrative data from short-term disability and prescription drug benefit claims and occupational health department records for employees of three Canadian companies. RESULTS: Approximately 58 percent of employees who were receiving depression-related short-term disability benefits had made at least one antidepressant claim. Employees who did not use antidepressants typically reported significantly fewer symptoms at baseline on average than those who did. About 91 percent of the employees who used antidepressants filled at least one prescription for a guideline-recommended first-line agent. Approximately 79 percent of antidepressant dosages reflected those suggested by the Canadian Network for Mood and Anxiety Treatment, and three timeframe indicators suggested that most patients used antidepressants within the recommended timeframes. CONCLUSIONS: The results of this study represent an important first step in exploring the question of how antidepressants are used among workers with depression-related disability. For the most part, these workers and those whose depression was more severe were more likely to obtain antidepressants. PMID- 12719506 TI - Community integration of elderly mentally ill persons in psychiatric hospitals and two types of residences. AB - OBJECTIVE: Deinstitutionalization policy in the Netherlands has given rise to two new living arrangements for elderly long-term psychiatric patients. Both involve accommodation in mainstream residential homes for elderly persons, either concentrated in a specialized care unit or dispersed throughout the facility. The authors studied the effectiveness of these two housing models for the community integration of such residents compared with accommodation in a psychiatric hospital. METHODS: Three subsamples were selected: 49 residents in six units of concentrated housing, 47 residents in 12 units of dispersed housing, and 78 patients in 24 psychiatric hospital units, for a total sample of 174 participants. These samples were compared in a quasi-experimental, posttest-only design that used four measures of community integration: amount of perceived influence over one's daily life, involvement in social activities, social network size, and frequency of visits received from members of the network. To adjust for differences in the populations, the hospital patients were matched to the residential home residents, and confounding factors were controlled for. RESULTS: Residential homes afforded more privacy, were closer to public services, and had a more diversified population than psychiatric hospitals. Participants in dispersed housing experienced more personal influence over their lives than did hospital patients. Concentrated-housing participants were less enterprising and had smaller social networks. The three groups did not differ in the frequency of visits received from network members. CONCLUSIONS: Community-integrated facilities do not necessarily imply community-integrated residents. Only dispersed-housing residences were an improvement over hospitals, and then solely in terms of residents' influence over their own daily lives. The advantage of the dispersed-housing model is that it resembles independent living while its institutional nature offers structure and protection. PMID- 12719507 TI - Behavioral health screening policies in Medicaid programs nationwide. AB - Under the Early and Periodic Screening, Diagnosis, and Treatment (EPSDT) mandate, states are required to screen Medicaid-insured children for mental health and substance use disorders. This national study found that states vary considerably in their policies. Nearly half the states (23 in total) have not addressed behavioral health issues in their EPSDT screening tools at all. More states have screening tools that address mental health than substance use disorders. Most states have created their own screening tools, which suggests discomfort with or a lack of awareness of the standard tools available. Screening policy options to increase behavioral health screening rates are discussed. PMID- 12719508 TI - Psychotropic medication expenditures for community-dwelling elderly persons. AB - Psychotropic medication expenditures for community-dwelling elderly persons in the United States were examined with the 1996 Medical Expenditure Panel Survey (MEPS). According to the MEPS, an estimated $1.07 billion US dollars was spent on psychotropic agents, and 53 percent of that was paid out of pocket. Antidepressants represented 61 percent of psychotropic expenditures. High expenditures were found for selective serotonin reuptake inhibitors ($469 million US dollars, or 44 percent of psychotropic expenditures), benzodiazepines ($241 million US dollars, or 22 percent), and tricyclic antidepressants ($119 million US dollars, or 11 percent). Estimates for psychotropic expenditures in 2002 totaled $1.41 billion US dollars after adjustment for inflation. Although psychotropic prescription costs were substantial, increased generic medication use and Medicare prescription plans could reduce the financial burden on elderly persons. PMID- 12719509 TI - Cost evaluation of risperidone compared with olanzapine. AB - This study evaluated costs associated with risperidone and olanzapine treatment for schizophrenia. Data were collected from the Department of Veterans Affairs computerized database nine months before and nine months after patients began continuous treatment with risperidone (N=23) or olanzapine (N=47). Both agents were associated with significant reductions in psychiatric hospitalization costs. Median increases in antipsychotic costs were significantly higher for patients treated with olanzapine ($1,892) than for those treated with risperidone ($733). Mean dosages were 3.5 mg per day for the risperidone group and 18 mg per day for the olanzapine group. Although both treatments were associated with similar reductions in costs of psychiatric inpatient and outpatient care, it was significantly less expensive to prescribe risperidone than olanzapine. PMID- 12719510 TI - Large data sets are powerful. PMID- 12719511 TI - Large data sets are powerful. PMID- 12719512 TI - Patients' attitudes toward antidepressants. PMID- 12719514 TI - Suicide and schizophrenia. PMID- 12719515 TI - Suicide, firearms, and data. PMID- 12719516 TI - Medical disorders among psychiatric patients. PMID- 12719518 TI - Bacteria are not too small for spatial sensing of chemical gradients: an experimental evidence. AB - By analyzing the chemotactic behavior of a recently described marine bacterial species, we provide experimental evidence that bacteria are not too small for sensing chemical gradients spatially. The bipolar flagellated vibrioid bacteria (typical size 2 x 6 microm) exhibit a unique motility pattern as they translate along as well as rotate around their short axis, i.e., the pathways of the cell poles describe a double helix. The natural habitat of the bacteria is characterized by steep oxygen gradients where they accumulate in a band at their preferred oxygen concentration of approximately 2 microM. Single cells leaving the band toward the oxic region typically return to the band within 16 s following a U-shaped track. A detailed analysis of the tracks reveals that the cells must be able to sense the oxygen gradient perpendicular to their swimming direction. Thus, they can detect oxygen gradients along a distance of approximately 5 microm corresponding to the extension of their long axis. The observed behavior can be explained by the presence of two independent sensor regions at either cell pole that modulate the rotation speed of the polar flagellar bundles, i.e., the flagellar bundle at the cell pole exposed to higher oxygen concentration is rotating faster than the other bundle. A mathematical model based on these assumptions reproduces the observed swimming behavior of the bacteria. PMID- 12719519 TI - The hepatitis C virus p7 protein forms an ion channel that is inhibited by long alkyl-chain iminosugar derivatives. AB - We show that hepatitis C virus (HCV) p7 protein forms ion channels in black lipid membranes. HCV p7 ion channels are inhibited by long-alkyl-chain iminosugar derivatives, which have antiviral activity against the HCV surrogate bovine viral diarrhea virus. HCV p7 presents a potential target for antiviral therapy. PMID- 12719520 TI - Genomewide transcriptional changes associated with genetic alterations and nutritional supplementation affecting tryptophan metabolism in Bacillus subtilis. AB - DNA microarrays comprising approximately 95% of the Bacillus subtilis annotated protein coding ORFs were deployed to generate a series of snapshots of genomewide transcriptional changes that occur when cells are grown under various conditions that are expected to increase or decrease transcription of the trp operon segment of the aromatic supraoperon. Comparisons of global expression patterns were made between cells grown in the presence of indole acrylic acid, a specific inhibitor of tRNA(Trp) charging; cells deficient in expression of the mtrB gene, which encodes the tryptophan-activated negative regulatory protein, TRAP; WT cells grown in the presence or absence of two or three of the aromatic amino acids; and cells harboring a tryptophanyl tRNA synthetase mutation conferring temperature sensitive tryptophan-dependent growth. Our findings validate expected responses of the tryptophan biosynthetic genes and presumed regulatory interrelationships between genes in the different aromatic amino acid pathways and the histidine biosynthetic pathway. Using a combination of supervised and unsupervised statistical methods we identified approximately 100 genes whose expression profiles were closely correlated with those of the genes in the trp operon. This finding suggests that expression of these genes is influenced directly or indirectly by regulatory events that affect or are a consequence of altered tryptophan metabolism. PMID- 12719521 TI - A molecular approach to comparative phylogeography of extant Malagasy lemurs. AB - The lemurs of Madagascar provide an excellent model for exploring evolutionary diversification. This study investigates genetic divergence among most extant lemur taxa in relation to potential geographical boundaries to gene flow. For this purpose, approximately 2,400 bp of mitochondrial DNA (part of the COIII gene; ND3, ND4L, and ND4 genes; and five tRNAs) were sequenced in a total of 131 lemurs from 5 families, 12 genera, 25 species, and 18 subspecies to reconstruct phylogenetic relationships among them. The comprehensive range of taxa makes this a particularly suitable molecular data set to examine lemur evolution. Those data clearly reveal that the Betsiboka River acts as an isolating barrier between populations of lemurs in north-western Madagascar. The Tsiribihina River similarly serves as a barrier to gene flow between northern and southern populations of lemurs in central western Madagascar, whereas the Mahavavy River does not seem to lead to genetic isolation of lemur populations. Several discrepancies among molecular data, current taxonomy, and geographic distribution along the western coast emerged. Examination of geographical distribution of the taxa concerned in comparison with distribution boundaries of other lemur taxa in that region yielded explanations for these inconsistencies. Eulemur fulvus and Eulemur mongoz are the only lemur taxa that also occur outside Madagascar, on the Comoro Islands. Genetic data show no significant differentiation between Malagasy and Comorian populations of these species, supporting the interpretation that both were introduced only recently to the Comoro Islands. PMID- 12719522 TI - Complex movements evoked by microstimulation of the ventral intraparietal area. AB - Most neurons in the ventral intraparietal area (VIP) of the macaque brain respond to both visual and tactile stimuli. The tactile receptive field is usually on the face, and the visual receptive field usually corresponds spatially to the tactile receptive field. In this study, electrical microstimulation of VIP, but not of surrounding tissue, caused a constellation of movements including eye closure, facial grimacing, head withdrawal, elevation of the shoulder, and movements of the hand to the space beside the head or shoulder. A similar set of movements was evoked by an air puff to the monkey's cheek. One interpretation is that VIP contributes to defensive movements triggered by stimuli on or near the head. PMID- 12719523 TI - Functional conservation of light, oxygen, or voltage domains in light sensing. AB - In Neurospora, the flavin adenine dinucleotide-containing protein WHITE COLLAR-1 is the blue-light photoreceptor for the circadian clock and other light responses. The putative chromophore-binding domain of WC-1, its light, oxygen, or voltage (LOV) domain, is similar to the LOV domains found in the plant phototropins, the Neurospora VIVID (VVD) protein, and the Arabidopsis FKF1 and its related proteins. Studies of the plant phototropins have identified 11 flavin contacting residues that are also conserved in the LOV domains of WC-1, VVD, and FKF1. In this study, by mutating the putative WC-1 flavin-binding sites, we show that these sites are important for the light function of the protein, suggesting that the WC-1 LOV domain adapts a structure similar to that of the phototropin LOV domains. By creating a Neurospora strain in which the LOV domain of WC-1 is swapped with that of VVD, we show that the LOV domain of VVD partially replaces the function of the WC-1 LOV domain, suggesting that VVD is a wc-dependent photoreceptor in Neurospora. Furthermore, we show that the Neurosporastrains containing a chimeric WC-1 protein with the LOV domain from FKF1 or phot1 can also sense light, suggesting that FKF1 and its related proteins are light sensors in Arabidopsis. Taken together, our data suggest that these LOV domains are structurally similar protein modules involved in blue-light sensing. PMID- 12719524 TI - Antigens in tea-beverage prime human Vgamma 2Vdelta 2 T cells in vitro and in vivo for memory and nonmemory antibacterial cytokine responses. AB - Human gammadelta T cells mediate innate immunity to microbes via T cell receptor dependent recognition of unprocessed antigens with conserved molecular patterns. These nonpeptide alkylamine antigens are shared by tumor cells, bacteria, parasites, and fungi but also by edible plant products such as tea, apples, mushrooms, and wine. Here we show that priming of gammadelta T cells with alkylamine antigens in vitro results in a memory response to these antigens. Such priming results also in a nonmemory response to whole bacteria and to lipopolysaccharide, characterized by IL-12-dependent secretion of IFN-gamma by gammadelta T cells and by gammadelta T cell proliferation. Drinking tea, which contains l-theanine, a precursor of the nonpeptide antigen ethylamine, primed peripheral blood gammadelta T cells to mediate a memory response on reexposure to ethylamine and to secrete IFN-gamma in response to bacteria. This unique combination of innate immune response and immunologic memory shows that gammadelta T cells can function as a bridge between innate and acquired immunity. In addition, these data provide an explanation for the health benefits of tea. PMID- 12719525 TI - Identification of genes involved in cell invasion by using a library of randomized hybrid ribozymes. AB - Libraries of RNA helicase-coupled randomized ribozymes are a powerful tool for the identification of functional genes. We have demonstrated the usefulness of this functional gene-discovery system by identifying genes involved in tumor invasion, a process that is an essential feature of tumor metastasis: the spread of cancer cells from the original tumor to other sites in the body that imposes serious problems in the prognosis and treatment of cancer. Using a filter-based invasion assay in vitro, we isolated ribozymes that enhanced the invasive properties of NIH 3T3 fibroblasts. Sequence analysis of selected clones and a database search revealed that genes such as the gene for Gem GTPase and uncharacterized genes that resemble genes for myosin phosphatase and protein tyrosine-phosphatase are involved in cell invasion. Our system for gene identification by using ribozymes and the functional analysis of target genes should help to clarify the complex mechanisms of invasion and metastasis and might provide information that is relevant to cancer therapy. PMID- 12719526 TI - The initiation-elongation transition: lateral mobility of RNA in RNA polymerase II complexes is greatly reduced at +8/+9 and absent by +23. AB - RNA polymerase II transcription complexes stalled shortly after initiation over a repetitive segment of the template can undergo efficient transcript slippage, during which the 3' end of the RNA slides upstream and then re-pairs with the template, allowing transcription to continue. In the present study, we have used transcript slippage as an assay to identify possible structural transitions that occur as the polymerase passes from the initiation to the elongation phase of transcription. We reasoned that transcript slippage would not occur in fully processive complexes. We constructed a series of templates that allowed us to stall RNA polymerase II after the synthesis of a repetitive sequence (5'-CUCUCU 3') at varying distances downstream of +1. We found that polymerase must synthesize at least a 23-nt RNA to attain resistance to transcript slippage. The ability to undergo slippage was lost in two discrete steps, suggestive of two distinct transitions. The first transition is the formation of the 8- to 9-bp mature RNA-DNA hybrid, when slippage abruptly dropped by 10-fold. However, easily detectable slippage continued until 14 more bonds were made. Thus, although the transcript becomes tightly constrained within the transcription complex once the hybrid reaches its final length, much more RNA synthesis is required before the RNA is no longer able to slip upstream along the template. This last point may reflect an important stabilizing role for the interaction of the polymerase with the transcript well upstream of the RNA-DNA hybrid. PMID- 12719527 TI - Long-term memory for a life on the move. AB - Evidence is accumulating that cognitive abilities are shaped by the specific ecological conditions to which animals are exposed. Long-distance migratory birds may provide a striking example of this. Field observations have shown that, at least in some species, a substantial proportion of individuals return to the same breeding, wintering, and stopover sites in successive years. This observation suggests that migrants have evolved special cognitive abilities that enable them to accomplish these feats. Here we show that memory of a particular feeding site persisted for at least 12 months in a long-distance migrant, whereas a closely related nonmigrant could remember such a site for only 2 weeks. Thus, it seems that the migratory lifestyle has influenced the learning and memorizing capacities of migratory birds. These results build a bridge between field observations suggesting special memorization feats of migratory birds and previous neuroanatomical results from the same two species indicating an increase in relative hippocampal size from the first to the second year of life in the migrant but not in the nonmigrant. PMID- 12719528 TI - Agonist unbinding from receptor dictates the nature of deactivation kinetics of G protein-gated K+ channels. AB - G protein-gated inwardly rectifying K(+) (Kir) channels are found in neurones, atrial myocytes, and endocrine cells and are involved in generating late inhibitory postsynaptic potentials, slowing the heart rate and inhibiting hormone release. They are activated by G protein-coupled receptors (GPCRs) via the inhibitory family of G protein, G(i/o), in a membrane-delimited fashion by the direct binding of Gbetagamma dimers to the channel complex. In this study we are concerned with the kinetics of deactivation of the cloned neuronal G protein gated K(+) channel, Kir3.1 + 3.2A, after stimulation of a number of GPCRs. Termination of the channel activity on agonist removal is thought to solely depend on the intrinsic hydrolysis rate of the G protein alpha subunit. In this study we present data that illustrate a more complex behavior. We hypothesize that there are two processes that account for channel deactivation: agonist unbinding from the GPCR and GTP hydrolysis by the G protein alpha subunit. With some combinations of agonist/GPCR, the rate of agonist unbinding is slow and rate limiting, and deactivation kinetics are not modulated by regulators of G protein signaling proteins. In another group, channel deactivation is generally faster and limited by the hydrolysis rate of the G protein alpha subunit. G protein isoform and interaction with G protein-signaling proteins play a significant role with this group of GPCRs. PMID- 12719529 TI - A TyrCD1/TrpG8 hydrogen bond network and a TyrB10TyrCD1 covalent link shape the heme distal site of Mycobacterium tuberculosis hemoglobin O. AB - Truncated hemoglobins (Hbs) are small hemoproteins, identified in microorganisms and in some plants, forming a separate cluster within the Hb superfamily. Two distantly related truncated Hbs, trHbN and trHbO, are expressed at different developmental stages in Mycobacterium tuberculosis. Sequence analysis shows that the two proteins share 18% amino acid identities and belong to different groups within the truncated Hb cluster. Although a specific defense role against nitrosative stress has been ascribed to trHbN (expressed during the Mycobacterium stationary phase), no clear functions have been recognized for trHbO, which is expressed throughout the Mycobacterium growth phase. The 2.1-A crystal structure of M. tuberculosis cyano-met trHbO shows that the protein assembles in a compact dodecamer. Six of the dodecamer subunits are characterized by a double conformation for their CD regions and, most notably, by a covalent bond linking the phenolic O atom of TyrB10 to the aromatic ring of TyrCD1, in the heme distal cavity. All 12 subunits display a cyanide ion bound to the heme Fe atom, stabilized by a tight hydrogen-bonded network based on the (globin very rare) TyrCD1 and TrpG8 residues. The small apolar AlaE7 residue leaves room for ligand access to the heme distal site through the conventional "E7 path," as proposed for myoglobin. Different from trHbN, where a 20-A protein matrix tunnel is held to sustain ligand diffusion to an otherwise inaccessible heme distal site, the topologically related region in trHbO hosts two protein matrix cavities. PMID- 12719530 TI - On the role of MHC class II molecules in the survival and lymphopenia-induced proliferation of peripheral CD4+ T cells. AB - CD4(+) T cells expand after transfer into lymphopenic H-2(b) A(beta)-/- mice (I A(beta)-, I-E(alpha)-deficient mice) but not after transfer into lymphopenic MHC II(Delta/Delta) mice (I-A(alpha)-, I-A(beta)-, I-E(alpha)-, and I-E(beta) deficient mice), implying that in H-2(b) A(beta)-/- mice, A(alpha) chain and E(beta) chain associate to form a hybrid A(alpha)E(beta) MHC class II molecule. In light of this unexpected result, we reexamined the MHC class II requirement in the survival and lymphopenia-induced proliferation of CD4(+) T cells. Here we show that expansion, but not short-term survival, of CD4(+) T cells depends on interactions with MHC class II molecules in lymphopenic mice. Nevertheless, interactions with classical MHC class II molecules are required for CD4(+) T cells to survive in CD8(+) T-cell-containing mice. PMID- 12719531 TI - Histone H1.2 is a substrate for denitrase, an activity that reduces nitrotyrosine immunoreactivity in proteins. AB - Several reports have described an activity that modifies nitrotyrosine-containing proteins and their immunoreactivity to nitrotyrosine Abs. Without knowing the product of the reaction, this new activity has been called a "denitrase." In those studies, some nonspecific proteins, which have multiple tyrosine residues, e.g., albumin, were used as a substrate. Therefore, the studies were based on an unknown mechanism of reaction and potentially a high background. To solve these problems, one of the most important things is to find a more suitable substrate for assay of the enzyme. We developed an assay strategy for determining the substrate for denitrase combining 2D-gel electrophoresis and an on-blot enzyme assay. The resulting substrate from RAW 264.7 cells was Histone H1.2, an isoform protein of linker histone. Histone H1.2 has only one tyrosine residue in the entire molecule, which ensures the exact position of the substrate to be involved. It has been reported that Histones are the most prominent nitrated proteins in cancer tissues. It was also demonstrated that tyrosine nitration of Histone H1 occurs in vivo. These findings lead us to the idea that Histone H1.2 might be an intrinsic substrate for denitrase. We nitrated recombinant and purified Histone H1.2 chemically and subjected it to an on-blot enzyme assay to characterize the activity. Denitrase activity behaved as an enzymatic activity because the reaction was time dependent and was destroyed by heat or trypsin treatment. The activity was shown to be specific for Histone H1.2, to differ from proteasome activity, and to require no additional cofactors. PMID- 12719532 TI - Sexual transmission of the [Het-S] prion leads to meiotic drive in Podospora anserina. AB - In the filamentous fungus Podospora anserina, two phenomena are associated with polymorphism at the het-s locus, vegetative incompatibility and ascospore abortion. Two het-s alleles occur naturally, het-s and het-S. The het-s encoded protein is a prion propagating as a self-perpetuating amyloid aggregate. When prion-infected [Het-s] hyphae fuse with [Het-S] hyphae, the resulting heterokaryotic cells necrotize. [Het-s] and [Het-S] strains are sexually compatible. When, however, a female [Het-s] crosses with [Het-S], a significant percentage of het-S spores abort, in a way similar to spore killing in Neurospora and Podospora. We report here that sexual transmission of the [Het-s] prion after nonisogamous mating in the reproductive cycle of Podospora is responsible for the killing of het-S spores. Progeny of crosses between isogenic strains with distinct wild-type or introduced, ectopic het-s/S alleles were cytologically and genetically analyzed. The effect of het-s/S overexpression, ectopic het-s/S expression, absence of het-s expression, loss of [Het-s] prion infection, and the distribution patterns of HET-s/S-GFP proteins were categorized during meiosis and ascospore formation. This study unveiled a het-S spore-killing system that is governed by dosage of and interaction between the [Het-s] prion and the HET-S protein. Due to this property of the [Het-s] prion, the het-s allele acts as a meiotic drive element favoring maintenance of the prion-forming allele in natural populations. PMID- 12719533 TI - Natural variation in human membrane transporter genes reveals evolutionary and functional constraints. AB - Membrane transporters maintain cellular and organismal homeostasis by importing nutrients and exporting toxic compounds. Transporters also play a crucial role in drug response, serving as drug targets and setting drug levels. As part of a pharmacogenetics project, we screened exons and flanking intronic regions for variation in a set of 24 membrane transporter genes (96 kb; 57% coding) in 247 DNA samples from ethnically diverse populations. We identified 680 single nucleotide polymorphisms (SNPs), of which 175 were synonymous and 155 caused amino acid changes, and 29 small insertions and deletions. Amino acid diversity (pi(NS)) in transmembrane domains (TMDs) was significantly lower than in loop domains, suggesting that TMDs have special functional constraints. This difference was especially striking in the ATP-binding cassette superfamily and did not parallel evolutionary conservation: there was little variation in the TMDs, even in evolutionarily unconserved residues. We used allele frequency distribution to evaluate different scoring systems (Grantham, blosum62, SIFT, and evolutionarily conservedevolutionarily unconserved) for their ability to predict which SNPs affect function. Our underlying assumption was that alleles that are functionally deleterious will be selected against and thus under represented at high frequencies and over represented at low frequencies. We found that evolutionary conservation of orthologous sequences, as assessed by evolutionarily conservedevolutionarily unconserved and SIFT, was the best predictor of allele frequency distribution and hence of function. European Americans had an excess of high frequency alleles in comparison to African Americans, consistent with a historic bottleneck. In addition, African Americans exhibited a much higher frequency of population specific medium-frequency alleles than did European Americans. PMID- 12719534 TI - Evolutionary conservation predicts function of variants of the human organic cation transporter, OCT1. AB - The organic cation transporter, OCT1, is a major hepatic transporter that mediates the uptake of many organic cations from the blood into the liver where the compounds may be metabolized or secreted into the bile. Because OCT1 interacts with a variety of structurally diverse organic cations, including clinically used drugs as well as toxic substances (e.g., N-methylpyridinium, MPP(+)), it is an important determinant of systemic exposure to many xenobiotics. To understand the genetic basis of extensive interindividual differences in xenobiotic disposition, we functionally characterized 15 protein-altering variants of the human liver organic cation transporter, OCT1, in Xenopus oocytes. All variants that reduced or eliminated function (OCT1-R61C, OCT1-P341L, OCT1 G220V, OCT1-G401S, and OCT1-G465R) altered evolutionarily conserved amino acid residues. In general, variants with decreased function had amino acid substitutions that resulted in more radical chemical changes (higher Grantham values) and were less evolutionarily favorable (lower blosum62 values) than variants that maintained function. A variant with increased function (OCT1-S14F) changed an amino acid residue such that the human protein matched the consensus of the OCT1 mammalian orthologs. Our results indicate that changes at evolutionarily conserved positions of OCT1 are strong predictors of decreased function and suggest that a combination of evolutionary conservation and chemical change might be a stronger predictor of function. PMID- 12719535 TI - Mammalian linker-histone subtypes differentially affect gene expression in vivo. AB - Posttranslational modifications and remodeling of nucleosomes are critical factors in the regulation of transcription. Higher-order folding of chromatin also is likely to contribute to the control of gene expression, but the absence of a detailed description of the structure of the chromatin fiber has impaired progress in this area. Mammalian somatic cells contain a set of H1 linker-histone subtypes, H1 (0) and H1a to H1e, that bind to nucleosome core particles and to the linker DNA between nucleosomes. To determine whether the H1 histone subtypes play differential roles in the regulation of gene expression, we combined mice lacking specific H1 histone subtypes with mice carrying transgenes subject to position effects. Because position effects result from the unique chromatin structure created by the juxtaposition of regulatory elements in the transgene and at the site of integration, transgenes can serve as exquisitely sensitive indicators of chromatin structure. We report that some, but not all, linker histones can attenuate or accentuate position effects. The results suggest that the linker-histone subtypes play differential roles in the control of gene expression and that the sequential arrangement of the linker histones on the chromatin fiber might regulate higher-order chromatin structure and fine-tune expression levels. PMID- 12719536 TI - Hydrogen-exchange stability analysis of Bergerac-Src homology 3 variants allows the characterization of a folding intermediate in equilibrium. AB - Amide hydrogendeuterium exchange rates have been determined for two mutants of alpha-spectrin Src homology 3 domain (WT), containing an elongated stable (SHH) and unstable (SHA) distal loop. SHA, similarly to WT, follows a two-state transition, whereas SHH apparently folds via a three-state mechanism. Native state amide hydrogen exchange is effective in ascribing energetic readjustments observed in kinetic experiments to species stabilized within the denatured base and distinguishing those from high-energy barrier crossings. Comparison of DeltaG(ex) and m(ex) parameters for amide protons of these mutants demonstrates the existence of an intermediate and allows the identification of protons protected in this state. The consolidation of a form containing a prefolded long beta-hairpin induces the switch to a three-state mechanism in an otherwise two state folder. It can be inferred that the unbalanced high stability of individual elements of secondary structure in a polypeptide could ultimately complicate its folding mechanism. PMID- 12719537 TI - Characterization of a family of endogenous neuropeptide ligands for the G protein coupled receptors GPR7 and GPR8. AB - GPR7 and GPR8 are orphan G protein-coupled receptors that are highly similar to each other. These receptors are expressed predominantly in brain, suggesting roles in central nervous system function. We have purified an endogenous peptide ligand for GPR7 from bovine hypothalamus extracts. This peptide, termed neuropeptide B (NPB), has a C-6-brominated tryptophan residue at the N terminus. It binds and activates human GPR7 or GPR8 with median effective concentrations (EC(50)) of 0.23 nM and 15.8 nM, respectively. In situ hybridization shows distinct localizations of the prepro-NPB mRNA in mouse brain, i.e., in paraventricular hypothalamic nucleus, hippocampus, and several nuclei in midbrain and brainstem. Intracerebroventricular (i.c.v.) injection of NPB in mice induces hyperphagia during the first 2 h, followed by hypophagia. Intracerebroventricular injection of NPB produces analgesia to s.c. formalin injection in rats. Through EST database searches, we identified a putative paralogous peptide. This peptide, termed neuropeptide W (NPW), also has an N-terminal tryptophan residue. Synthetic human NPW binds and activates human GPR7 or GPR8 with EC(50) values of 0.56 nM and 0.51 nM, respectively. The expression of NPW mRNA in mouse brain is confined to specific nuclei in midbrain and brainstem. These findings suggest diverse physiological functions of NPB and NPW in the central nervous system, acting as endogenous ligands on GPR7 andor GPR8. PMID- 12719538 TI - Adeno-associated viral transfer of opioid receptor gene to primary sensory neurons: a strategy to increase opioid antinociception. AB - To develop a genetic approach for the treatment of pain, we introduced a recombinant adeno-associated viral (rAAV) vector containing the cDNA for the mu opioid receptor (muOR) into primary afferent neurons in dorsal root ganglia (DRGs) of rats, which resulted in a long-lasting (>6 months) increase in muOR expression in DRG neurons. The increase greatly potentiated the antinociceptive effects of morphine in rAAV-muOR-infected rats with and without inflammation. Perforated patch recordings indicated that the efficacy and potency of opioid inhibition of voltage-dependent Ca(2+) channels were enhanced in infected neurons, which may underlie the increase in opiate efficacy. These data suggest that transfer of opioid receptor genes into DRG cells with rAAV vectors may offer a new therapeutic strategy for pain management. PMID- 12719539 TI - Parkin, a gene implicated in autosomal recessive juvenile parkinsonism, is a candidate tumor suppressor gene on chromosome 6q25-q27. AB - In an effort to identify tumor suppressor gene(s) associated with the frequent loss of heterozygosity observed on chromosome 6q25-q27, we constructed a contig derived from the sequences of bacterial artificial chromosomeP1 bacteriophage artificial chromosome clones defined by the genetic interval D6S1581-D6S1579 D6S305-D6S1599-D6S1008. Sequence analysis of this contig found it to contain eight known genes, including the complete genomic structure of the Parkin gene. Loss of heterozygosity (LOH) analysis of 40 malignant breast and ovarian tumors identified a common minimal region of loss, including the markers D6S305 (50%) and D6S1599 (32%). Both loci exhibited the highest frequencies of LOH in this study and are each located within the Parkin genomic structure. Whereas mutation analysis revealed no missense substitutions, expression of the Parkin gene appeared to be down-regulated or absent in the tumor biopsies and tumor cell lines examined. In addition, the identification of two truncating deletions in 3 of 20 ovarian tumor samples, as well as homozygous deletion of exon 2 in the lung adenocarcinoma cell lines Calu-3 and H-1573, supports the hypothesis that hemizygous or homozygous deletions are responsible for the abnormal expression of Parkin in these samples. These data suggest that the LOH observed at chromosome 6q25-q26 may contribute to the initiation andor progression of cancer by inactivating or reducing the expression of the Parkin gene. Because Parkin maps to FRA6E, one of the most active common fragile sites in the human genome, it represents another example of a large tumor suppressor gene, like FHIT and WWOX, located at a common fragile site. PMID- 12719540 TI - Efficient linking and transfer of multiple genes by a multigene assembly and transformation vector system. AB - Introduction and expression of multiple transgenes is frequently required for basic and applied studies. However, at present, multigene transformation is very difficult due to technical limitations of existing methods. Here, we describe a vector system for efficient multigene assembly and transformation. The system consists of a transformation-competent artificial chromosome (TAC)-based acceptor vector together with two donor vectors. By exploiting the CreloxP recombination system and homing endonucleases, multiple rounds of gene assembly cycling were carried out with alternate use of the donor vectors, and multiple genes were sequentially delivered into the TAC vector. With this system, we created constructs containing as many as 10 foreign DNA fragments. Multiple genes, including six resistant genes stacked in a construct, were transferred into rice genome by Agrobacterium-mediated transformation. This system extends the repertoire of molecular genetic studies and biotechnological endeavors by enabling simultaneous manipulation of multiple genes. PMID- 12719541 TI - Cloning of an Alpha-TFEB fusion in renal tumors harboring the t(6;11)(p21;q13) chromosome translocation. AB - MITF, TFE3, TFEB, and TFEC comprise a transcription factor family (MiT) that regulates key developmental pathways in several cell lineages. Like MYC, MiT members are basic helix-loop-helix-leucine zipper transcription factors. MiT members share virtually perfect homology in their DNA binding domains and bind a common DNA motif. Translocations of TFE3 occur in specific subsets of human renal cell carcinomas and in alveolar soft part sarcomas. Although multiple translocation partners are fused to TFE3, each translocation product retains TFE3's basic helix-loop-helix leucine zipper. We have identified the genes fused by the chromosomal translocation t(6;11)(p21.1;q13), characteristic of another subset of renal neoplasms. In two primary tumors we found that Alpha, an intronless gene, rearranges with the first intron of TFEB, just upstream of TFEB's initiation ATG, preserving the entire TFEB coding sequence. Fluorescence in situ hybridization confirmed the involvement of both TFEB and Alpha in this translocation. Although the Alpha promoter drives expression of this fusion gene, the Alpha gene does not contribute to the ORF. Whereas TFE3 is typically fused to partner proteins in subsets of renal tumors, we found that wild-type, unfused TFE3 stimulates clonogenic growth in a cell-based assay, suggesting that dysregulated expression, rather than altered function of TFEB or TFE3 fusions, may confer neoplastic properties, a mechanism reminiscent of MYC activation by promoter substitution in Burkitt's lymphoma. Alpha-TFEB is thus identified as a fusion gene in a subset of pediatric renal neoplasms. PMID- 12719542 TI - An unusual mechanism of bacterial gene expression revealed for the RNase P protein of Thermus strains. AB - The RNase P protein gene (rnpA) completely overlaps the rpmH gene (encoding ribosomal protein L34) out of frame in the thermophilic bacterium Thermus thermophilus. This results in the synthesis of an extended RNase P protein (C5) of 163 aa and, by inference, of 240 aa in the related strain Thermus filiformis. Start codons of rnpA and rpmH, apparently governed by the same ribosome binding site, are separated by only 4 nt, which suggests a regulatory linkage between L34 and C5 translation and, accordingly, between ribosome and RNase P biosynthesis. Within the sequence encoding the N-terminal extensions and downstream of rpmH, several Thermus species exhibit in-frame deletionsinsertions, suggesting relaxed constraints for sequence conservation in this region. Roughly the N-terminal third of T. thermophilus C5 was further shown to be dispensable for RNase P function in vitro by using a precursor tRNA(Gly) substrate from the same organism. Taken together, these data reveal a mode of gene expression that is to our knowledge unprecedented in bacteria. PMID- 12719543 TI - Different time courses for visual perception and action priming. AB - Visual stimuli may remain invisible but nevertheless produce strong and reliable effects on subsequent actions. How well features of a masked prime are perceived depends crucially on its physical parameters and those of the mask. We manipulated the visibility of masked stimuli and contrasted it with their influence on the speed of motor actions, comparing the temporal dynamics of visual awareness in metacontrast masking with that of action priming under the same conditions. We observed priming with identical time course for reportable and invisible prime stimuli, despite qualitative changes in the masking time course. Our findings indicate that experimental variations that modify the subjective visual experience of masked stimuli have no effect on motor effects of those stimuli in early processing. We propose a model that provides a quantitative account of priming effects on response speed and accuracy. PMID- 12719545 TI - Tropical rain forest tree growth and atmospheric carbon dynamics linked to interannual temperature variation during 1984-2000. AB - During 1984-2000, canopy tree growth in old-growth tropical rain forest at La Selva, Costa Rica, varied >2-fold among years. The trees' annual diameter increments in this 16-yr period were negatively correlated with annual means of daily minimum temperatures. The tree growth variations also negatively covaried with the net carbon exchange of the terrestrial tropics as a whole, as inferred from nearly pole-to-pole measurements of atmospheric carbon dioxide (CO(2)) interpreted by an inverse tracer-transport model. Strong reductions in tree growth and large inferred tropical releases of CO(2) to the atmosphere occurred during the record-hot 1997-1998 El Nino. These and other recent findings are consistent with decreased net primary production in tropical forests in the warmer years of the last two decades. As has been projected by recent process model studies, such a sensitivity of tropical forest productivity to on-going climate change would accelerate the rate of atmospheric CO(2) accumulation. PMID- 12719544 TI - A Wnt- and beta -catenin-dependent pathway for mammalian cardiac myogenesis. AB - Acquisition of a cardiac fate by embryonic mesodermal cells is a fundamental step in heart formation. Heart development in frogs and avians requires positive signals from adjacent endoderm, including bone morphogenic proteins, and is antagonized by a second secreted signal, Wnt proteins, from neural tube. By contrast, mechanisms of mesodermal commitment to create heart muscle in mammals are largely unknown. In addition, Wnt-dependent signals can involve either a canonical beta-catenin pathway or other, alternative mediators. Here, we tested the involvement of Wnts and beta-catenin in mammalian cardiac myogenesis by using a pluripotent mouse cell line (P19CL6) that recapitulates early steps for cardiac specification. In this system, early and late cardiac genes are up-regulated by 1% DMSO, and spontaneous beating occurs. Notably, Wnt3A and Wnt8A were induced days before even the earliest cardiogenic transcription factors. DMSO induced biochemical mediators of Wnt signaling (decreased phosphorylation and increased levels of beta-catenin), which were suppressed by Frizzled-8Fc, a soluble Wnt antagonist. DMSO provoked T cell factor-dependent transcriptional activity; thus, induction of Wnt proteins by DMSO was functionally coupled. Frizzled-8Fc inhibited the induction of cardiogenic transcription factors, cardiogenic growth factors, and sarcomeric myosin heavy chains. Likewise, differentiation was blocked by constitutively active glycogen synthase kinase 3beta, an intracellular inhibitor of the Wntbeta-catenin pathway. Conversely, lithium chloride, which inhibits glycogen synthase kinase 3beta, and Wnt3A-conditioned medium up regulated early cardiac markers and the proportion of differentiated cells. Thus, Wntbeta-catenin signaling is activated at the inception of mammalian cardiac myogenesis and is indispensable for cardiac differentiation, at least in this pluripotent model system. PMID- 12719546 TI - Change of conduction velocity by regional myelination yields constant latency irrespective of distance between thalamus and cortex. AB - The widely spanning sensory cortex receives inputs from the disproportionately smaller nucleus of the thalamus, which results in a wide variety of travelling distance among thalamic afferents. Yet, latency from the thalamus to a cortical cell is remarkably constant across the cortex (typically, approximately 2 ms). Here, we found a mechanism that produces invariability of latency among thalamocortical afferents, irrespective of the variability of travelling distances. The conduction velocity (CV) was calculated from excitatory postsynaptic currents recorded from layer IV cells in mouse thalamocortical slices by stimulating the ventrobasal nucleus of the thalamus (VB) and white matter (WM). In adults, the obtained CV for VB to WM (CV(VB-WM); 3.28 +/- 0.11 ms) was approximately 10 times faster than that of WM to layer IV cells (CV(WM IV); 0.33 +/- 0.05 ms). The CV(VB-WM) was confirmed by recording antidromic single-unit responses from VB cells by stimulating WM. Exclusion of synaptic delay from CV(WM-IV) did not account for the 10-fold difference of CV. By histochemical staining, it was revealed that VB to WM was heavily myelinated, whereas in the cortex staining became substantially weaker. We also found that such morphological and physiological characteristics developed in parallel and were accomplished around postnatal week 4. Considering that VB to WM is longer and more variable in length among afferents than is the intracortical region, such an enormous difference of CV makes conduction time heavily dependent on the length of intracortical region, which is relatively constant. Our finding may well provide a general strategy of connecting multiple sites irrespective of distances in the brain. PMID- 12719548 TI - Role of murine cytomegalovirus US22 gene family members in replication in macrophages. AB - The large cytomegalovirus (CMV) US22 gene family, found in all betaherpesviruses, comprises 12 members in both human cytomegalovirus (HCMV) and murine cytomegalovirus (MCMV). Conserved sequence motifs suggested a common ancestry and related functions for these gene products. Two members of this family, m140 and m141, were recently shown to affect MCMV replication on macrophages. To test the role of all US22 members in cell tropism, we analyzed the growth properties in different cell types of MCMV mutants carrying transposon insertions in all 12 US22 gene family members. When necessary, additional targeted mutants with gene deletions, ATG deletions, and ectopic gene revertants were constructed. Mutants with disruption of genes M23, M24, m25.1, m25.2, and m128 (ie2) showed no obvious growth phenotype, whereas growth of M43 mutants was reduced in a number of cell lines. Genes m142 and m143 were shown to be essential for virus replication. Growth of mutants with insertions into genes M36, m139, m140, and m141 in macrophages was severely affected. The common phenotype of the m139, m140, and m141 mutants was explained by an interaction at the protein level. The M36 dependent macrophage growth phenotype could be explained by the antiapoptotic function of the gene that was required for growth on macrophages but not for growth on other cell types. Together, the comprehensive set of mutants of the US22 gene family suggests that individual family members have diverged through evolution to serve a variety of functions for the virus. PMID- 12719547 TI - Elimination of protease activity restores efficient virion production to a human immunodeficiency virus type 1 nucleocapsid deletion mutant. AB - The nucleocapsid (NC) region of human immunodeficiency virus type 1 (HIV-1) Gag is required for specific genomic RNA packaging. To determine if NC is absolutely required for virion formation, we deleted all but seven amino acids from NC in a full-length NL4-3 proviral clone. This construct, DelNC, produced approximately four- to sixfold fewer virions than did the wild type, and these virions were noninfectious (less than 10(-6) relative to the wild type) and severely genomic RNA deficient. Immunoblot and high-pressure liquid chromatography analyses showed that all of the mature Gag proteins except NC were present in the mutant virion preparations, although there was a modest decrease in Gag processing. DelNC virions had lower densities and were more heterogeneous than wild-type particles, consistent with a defect in the interaction assembly or I domain. Electron microscopy showed that the DelNC virions displayed a variety of aberrant morphological forms. Inactivating the protease activity of DelNC by mutation or protease inhibitor treatment restored virion production to wild-type levels. DelNC-protease mutants formed immature-appearing particles that were as dense as wild-type virions without incorporating genomic RNA. Therefore, protease activity combined with the absence of NC causes the defect in DelNC virion production, suggesting that premature processing of Gag during assembly causes this effect. These results show that HIV-1 can form particles efficiently without NC. PMID- 12719549 TI - African Swine Fever virus proteinase is essential for core maturation and infectivity. AB - African swine fever virus (ASFV) encodes two polyprotein precursors named pp220 and pp62 that are sequentially processed during viral infection, giving rise to six major structural proteins. These reside at the core shell, a matrix domain located between the endoplasmic reticulum-derived inner envelope and the DNA containing nucleoid. Proteolytic processing of the polyprotein precursors is catalyzed by the viral proteinase pS273R, a cysteine proteinase that shares sequence similarity with the SUMO1-processing peptidases. We describe here the construction and characterization of an ASFV recombinant, vS273Ri, that inducibly expresses the ASFV proteinase. Using vS273Ri, we show that repression of proteinase expression inhibits polyprotein processing and strongly impairs infective virus production. Electron microscopic examination of vS273Ri-infected cells showed that inhibition of proteolytic processing leads to the assembly of defective icosahedral particles containing a noncentered electron-dense nucleoid surrounded by an abnormal core shell of irregular thickness. The analysis of purified extracellular defective particles revealed that they contain the unprocessed pp220 and pp62 precursors, as well as the major DNA-binding nucleoid proteins p10 and pA104R. Altogether, these results indicate that the proteolytic processing of the polyproteins is not required for their incorporation into the assembling particles nor for the incorporation of the DNA-containing nucleoid. Instead, the ASFV proteinase is involved in a late maturational step that is essential for proper core assembly and infectivity. PMID- 12719550 TI - Kaposi's sarcoma-associated herpesvirus lytic origin (ori-Lyt)-dependent DNA replication: identification of the ori-Lyt and association of K8 bZip protein with the origin. AB - Herpesviruses utilize different origins of replication during lytic versus latent infection. Latent DNA replication depends on host cellular DNA replication machinery, whereas lytic cycle DNA replication requires virally encoded replication proteins. In lytic DNA replication, the lytic origin (ori-Lyt) is bound by a virus-specified origin binding protein (OBP) that recruits the core replication machinery. In this report, we demonstrated that DNA sequences in two noncoding regions of the Kaposi's sarcoma-associated herpesvirus (KSHV) genome, between open reading frames (ORFs) K4.2 and K5 and between K12 and ORF71, are able to serve as origins for lytic cycle-specific DNA replication. The two ori Lyt domains share an almost identical 1,153-bp sequence and a 600-bp downstream GC-rich repeat sequence, and the 1.7-kb DNA sequences are sufficient to act as a cis signal for replication. We also showed that an AT-palindromic sequence in the ori-Lyt domain is essential for the DNA replication. In addition, a virally encoded bZip protein, namely K8, was found to bind to a DNA sequence within the ori-Lyt by using a DNA binding site selection assay. The binding of K8 to this region was confirmed in cells by using a chromatin immunoprecipitation method. Further analysis revealed that K8 binds to an extended region, and the entire region is 100% conserved between two KSHV ori-Lyt's. K8 protein displays significant similarity to the Zta protein of Epstein-Barr virus (EBV), which is a known OBP of EBV. This notion, together with the ability of K8 to bind to the KSHV ori-Lyt, suggests that K8 may function as an OBP in KSHV. PMID- 12719551 TI - Intranasal vaccination using interleukin-12 and cholera toxin subunit B as adjuvants to enhance mucosal and systemic immunity to human immunodeficiency virus type 1 glycoproteins. AB - We have investigated the induction of protective mucosal immunity to human immunodeficiency virus type 1 (HIV-1) isolate 89.6 by intranasal (i.n.) immunization of mice with gp120 and gp140 together with interleukin-12 (IL-12) and cholera toxin subunit B (CTB) as adjuvants. It was found that both IL-12 and CTB were required to elicit mucosal antibody responses and that i.n. immunization resulted in increased total, immunoglobulin G1 (IgG1), and IgG2a anti-HIV-1 antibody levels in serum; increased total, IgG1, IgG2a, and IgA antibody expression in bronchoalveolar lavage fluids; and increased IgA antibody levels in vaginal washes. Levels of anti-HIV-1 antibodies in both sera and secretions were higher in groups immunized with gp140 than in those immunized with gp120. However, only gp120-specific mucosal antibodies demonstrated neutralizing activity against HIV-1 89.6. Taken together, the results show that IL-12 and CTB act synergistically to enhance both systemic and local mucosal antibody responses to HIV-1 glycoproteins and that even though gp140 induces higher antibody titers than gp120, only gp120-specific mucosal antibodies interfere with virus infectivity. PMID- 12719552 TI - Sindbis virus vectors designed to express a foreign protein as a cleavable component of the viral structural polyprotein. AB - Alphavirus-based expression vectors commonly use a duplicated 26S promoter to drive expression of a foreign gene. Here we describe an expression strategy in which the foreign sequences are linked to the gene encoding the 2A protease of foot-and-mouth disease virus and then inserted in frame between the capsid and E3 genes of Sindbis virus. During replication, the 2A fusion protein is synthesized as a component of the viral structural polyprotein that is then released by intramolecular cleavages mediated by the capsid and 2A proteases. Recombinant Sindbis viruses that expressed fusion proteins composed of 2A linked to the green fluorescent protein (GFP) and to the VP7 protein of bluetongue virus were constructed. Viruses engineered to express GFP and VP7 from a duplicate 26S promoter were also constructed. All four viruses expressed the transgene and grew to similar titers in cultured cells. However, the GFP/2A- and VP7/2A-expressing viruses displayed greater expression stability and were less attenuated in newborn mice than the cognate double-subgenomic promoter-based viruses. By combining the two expression strategies, we constructed bivalent viruses that incorporated and expressed both transgenes. The bivalent viruses grew to lower titers in cultured cells and were essentially avirulent in newborn mice. Groups of mice were vaccinated with each VP7- and VP7/2A-expressing virus, and antibody responses to native VP7 were measured in an indirect enzyme-linked immunosorbent assay. Despite their genetic and phenotypic differences, all viruses induced similarly high titers of VP7-specific antibodies. These results demonstrate that 2A fusion protein-expressing alphaviruses may be particularly well suited for applications that require enduring expression of a single protein or coexpression of two alternative proteins. PMID- 12719553 TI - Mutational analysis of open reading frames 62 and 71, encoding the varicella zoster virus immediate-early transactivating protein, IE62, and effects on replication in vitro and in skin xenografts in the SCID-hu mouse in vivo. AB - The varicella-zoster virus (VZV) genome has unique long (U(L)) and unique short (U(S)) segments which are flanked by internal repeat (IR) and terminal repeat (TR) sequences. The immediate-early 62 (IE62) protein, encoded by open reading frame 62 (ORF62) and ORF71 in these repeats, is the major VZV transactivating protein. Mutational analyses were done with VZV cosmids generated from parent Oka (pOka), a low-passage clinical isolate, and repair experiments were done with ORF62 from pOka and vaccine Oka (vOka), which is derived from pOka. Transfections using VZV cosmids from which ORF62, ORF71, or the ORF62/71 gene pair was deleted showed that VZV replication required at least one copy of ORF62. The insertion of ORF62 from pOka or vOka into a nonnative site in U(S) allowed VZV replication in cell culture in vitro, although the plaque size and yields of infectious virus were decreased. Targeted mutations in binding sites reported to affect interaction with IE4 protein and a putative ORF9 protein binding site were not lethal. Single deletions of ORF62 or ORF71 from cosmids permitted recovery of infectious virus, but recombination events repaired the defective repeat region in some progeny viruses, as verified by PCR and Southern hybridization. VZV infectivity in skin xenografts in the SCID-hu model required ORF62 expression; mixtures of single-copy recombinant Oka Delta 62 (rOka Delta 62) or rOka Delta 71 and repaired rOka generated by recombination of the single-copy deletion mutants were detected in some skin implants. Although insertion of ORF62 into the nonnative site permitted replication in cell culture, ORF62 expression from its native site was necessary for cell-cell spread in differentiated human skin tissues in vivo. PMID- 12719554 TI - Trafficking of human immunodeficiency virus type 1-specific CD8+ T cells to gut associated lymphoid tissue during chronic infection. AB - Gut-associated lymphoid tissue (GALT) is a significant but understudied lymphoid organ, harboring a majority of the body's total lymphocyte population. GALT is also an important portal of entry for human immunodeficiency virus (HIV), a major site of viral replication and CD4(+) T-cell depletion, and a frequent site of AIDS-related opportunistic infections and neoplasms. However, little is known about HIV-specific cell-mediated immune responses in GALT. Using lymphocytes isolated from rectal biopsies, we have determined the frequency and phenotype of HIV-specific CD8(+) T cells in human GALT. GALT CD8(+) T cells were predominantly CD45RO(+) and expressed CXCR4 and CCR5. In 10 clinically stable, chronically infected individuals, the frequency of HIV Gag (SL9)-specific CD8(+) T cells was increased in GALT relative to peripheral blood mononuclear cells by up to 4.6 fold, while that of cytomegalovirus (CMV)-specific CD8(+) T cells was significantly reduced (P = 0.012). Both HIV- and CMV-specific CD8(+) T cells in GALT expressed CCR5, but only HIV-specific CD8(+) T cells expressed alpha E beta 7 integrin, suggesting that mucosal priming may account for their retention in GALT. Chronically infected individuals exhibited striking depletion of GALT CD4(+) T cells expressing CXCR4, CCR5, and alpha E beta 7 integrin, but CD4(+)/CD8(+) T-cell ratios in blood and GALT were similar. The percentage of GALT CD8(+) T cells expressing alpha E beta 7 was significantly decreased in infected individuals, suggesting that HIV infection may perturb lymphocyte retention in GALT. These studies demonstrate the feasibility of using tetramers to assess HIV-specific T cells in GALT and reveal that GALT is the site of an active CD8(+) T-cell response during chronic infection. PMID- 12719555 TI - Tmevpg1, a candidate gene for the control of Theiler's virus persistence, could be implicated in the regulation of gamma interferon. AB - The Tmevp3 locus controls the load of Theiler's virus RNA during persistent infection of the mouse central nervous system (CNS). We identified a candidate gene at this locus, Tmevpg1, by using a positional cloning approach. Tmevpg1 and its human ortholog, TMEVPG1, are expressed in the immune system and encode what appears to be a noncoding RNA. They are located in a cluster of cytokine genes that includes the genes for gamma interferon and one or two homolog of interleukin-10. We now report that Tmevpg1 is expressed in CNS-infiltrating immune cells of resistant B10.S mice, but not in those of susceptible SJL/J mice, following inoculation with Theiler's virus. The pattern of expression of Tmevpg1 is the same in B10.S mice and in SJL/J mice congenic for the resistant B10.S haplotype of Tmevp3. Nineteen polymorphisms were identified when the Tmevpg1 genes of B10.S and SJL/J mice were compared. Interestingly, Tmevpg1 is down regulated after in vitro stimulation of murine CD4(+) or CD8(+) splenocytes, whereas Ifng is up regulated. Similar patterns of expression of TMEVPG1 and IFNG were observed in human NK cells and CD4(+) and CD8(+) T lymphocytes. Therefore, Tmevpg1 is a strong candidate gene for the Tmevp3 locus and may be involved in the control of Ifng gene expression. PMID- 12719556 TI - Transcriptional regulatory properties of Epstein-Barr virus nuclear antigen 3C are conserved in simian lymphocryptoviruses. AB - Epstein-Barr virus (EBV) nuclear antigen 3C (EBNA-3C) is a large transcriptional regulator essential for EBV-mediated immortalization of B lymphocytes. We previously identified interactions between EBNA-3C and two cellular transcription factors, J kappa and Spi proteins, through which EBNA-3C regulates transcription. To better understand the contribution of these interactions to EBNA-3C function and EBV latency, we examined whether they are conserved in the homologous proteins of nonhuman primate lymphocryptoviruses (LCVs), which bear a strong genetic and biological similarity to EBV. The homologue of EBNA-3C encoded by the LCV that infects baboons (BaLCV) was found to be only 35% identical in sequence to its EBV counterpart. Of particular significance, this homology localized predominantly to the N-terminal half of the molecule, which encompasses the domains in EBNA-3C that interact with J kappa and Spi proteins. Like EBNA-3C, both BaLCV and rhesus macaque LCV (RhLCV) 3C proteins bound to J kappa and repressed transcription mediated by EBNA-2 through its interaction with J kappa. Both nonhuman primate 3C proteins were also able to activate transcription mediated by the Spi proteins in the presence of EBNA-2. Like EBNA-3C, a domain encompassing the putative basic leucine zipper motif of the BaLCV-3C protein directly interacted with both Spi-1 and Spi-B. Surprisingly, a recently identified motif in EBNA-3C that mediates repression was not identifiable in the BaLCV-3C protein. Finally, although the C terminus of BaLCV-3C bears minimal homology to EBNA-3C, it nonetheless contains a C-terminal domain rich in glutamine and proline that was able to function as a potent transcriptional activation domain, as does the C terminus of EBNA-3C. The conservation of these functional motifs despite poor overall homology among the LCV 3C proteins strongly suggests that the interactions of EBNA-3C with J kappa and Spi do indeed play significant roles in the life cycle of EBV. PMID- 12719557 TI - Effect of p38 mitogen-activated protein kinase on the replication of encephalomyocarditis virus. AB - Cellular phosphorylation events during viral infection are necessary for effective viral replication. Encephalomyocarditis (EMC) virus has been used for studies on the molecular mechanisms of viral replication, but little is known about the cellular signaling pathways involved. This investigation was initiated to determine whether mitogen-activated protein kinases (MAPKs), which are central components of signal transduction pathways in the regulation of cell proliferation, play a role in the replication of EMC virus. We examined the phosphorylation of MAPKs, including extracellular signal-regulated kinase (ERK1/2), p38 MAPK, and stress-activated protein kinase 1/c-Jun NH(2)-terminal kinase (SAPK/JNK) in EMC virus-infected L929 cells and found that p38 MAPK and SAPK-JNK, but not ERK1/2, were activated during viral infection. We then examined the effect of these kinases on the replication of EMC virus in L929 cells by using specific inhibitors, including genistein or herbimycin A for tyrosine kinase, SB203580 or SB202190 for p38 MAPK, and PD98059 for ERK1/2. We found that the tyrosine kinase and p38 MAPK inhibitors, but not the ERK1/2 inhibitor, suppressed viral replication and that the inhibitory effect was primarily on viral protein synthesis. Finally, we examined whether p38 MAPK is involved in the translation of EMC viral transcripts by using L929 cells transfected with a gene construct containing the internal ribosomal entry site (IRES) of EMC virus and a luciferase reporter gene. We found that the p38 MAPK inhibitor suppressed the translation of EMC viral RNA. On the basis of these observations, we conclude that p38 MAPK plays a critical role in the replication of EMC virus, probably in the translation of viral RNA. PMID- 12719558 TI - L-particle production during primary replication of pseudorabies virus in the nasal mucosa of swine. AB - Different tissue culture cell lines infected with a number of alphaherpesviruses produce, in addition to virions, light particles (L particles). L particles are composed of the envelope and tegument components of the virion but totally lack the proteins of the capsid and the virus genome; therefore, they are noninfectious. In this electron microscopy report, we show that L particles are produced during primary replication of the alphaherpesvirus pseudorabies virus (PRV) in the nasal mucosa of experimentally infected swine, its natural host. Although PRV infected different types of cells of the respiratory and olfactory mucosae, PRV L particles were found to be produced exclusively by epithelial cells and fibroblasts. We observed that formation of noninfectious particles occurred by budding of condensed tegument at the inner nuclear membrane and at membranes of cytoplasmic vesicles, resulting in intracisternal and intravesicular L particles, respectively. Both forms of capsidless particles were clearly distinguishable by the presence of prominent surface projections on the envelope and the higher electron density of the tegument, morphological features which were only observed in intravesicular L particles. Moreover, intravesicular but not intracisternal L particles were found to be released by exocytosis and were also identified extracellularly. Comparative analysis between PRV virion and L particle morphogenesis indicates that both types of virus particles share a common intracellular pathway of assembly and egress but that they show different production patterns during the replication cycle of PRV. PMID- 12719559 TI - The highly conserved basic domain I of baculovirus IE1 is required for hr enhancer DNA binding and hr-dependent transactivation. AB - The immediate-early protein IE1 is the principal transcriptional regulator of the baculovirus Autographa californica nucleopolyhedrovirus (AcMNPV). Transactivation by IE1 is dramatically stimulated by cis linkage of the affected promoter to AcMNPV homologous region (hr) elements that contain palindromic 28-bp repeats (28 mers) with enhancer activity. This hr-dependent transcriptional enhancement requires binding of the 28-mer by dimeric IE1. Here, we have defined IE1 domains required for this DNA binding in order to investigate the mechanism of IE1 function. Analysis of a panel of IE1 insertion mutations indicated that disruption of a highly conserved domain (residues 152 to 161) consisting of mostly positive-charged residues (basic domain I) abolished hr-dependent transactivation. Targeted mutagenesis of basic residues within basic domain I caused loss of hr-dependent transactivation but had no effect on IE1 oligomerization, nuclear localization, or hr-independent transactivation of viral promoters. Alanine substitutions of K(152) and K(154) or K(160) and K(161) impaired IE1 binding to 28-mer DNA as a homodimer, indicating that these basic residues are required for enhancer binding. Consistent with a DNA-binding defect, 28-mer interaction was improved by heterodimerization with wild-type IE1 or by increasing mutated IE1 concentrations. DNA binding mediated by basic domain I was also required for IE1 transactivation that occurred through physically separated, unlinked hr elements. We concluded that basic domain I is the enhancer-binding domain for IE1. Our data also suggest that DNA binding activates IE1 for transcriptional enhancement, possibly through a conformational change involving basic domain I. PMID- 12719561 TI - A novel genetic pathway of human immunodeficiency virus type 1 resistance to stavudine mediated by the K65R mutation. AB - Stavudine (d4T) and zidovudine (AZT) are thymidine analogs widely used in the treatment of human immunodeficiency virus type 1 (HIV-1)-infected persons. Resistance to d4T is not fully understood, although the selection of AZT resistance mutations in patients treated with d4T suggests that both drugs have similar pathways of resistance. Through the analysis of genotypic changes in nine recombinant viruses cultured with d4T, we identified a new pathway for d4T resistance mediated by K65R, a mutation not selected by AZT. Passaged viruses were derived from treatment-naive persons or HIV-1(HXB2) and had wild-type reverse transcriptase (RT) or T215C/D mutations. K65R was selected in seven viruses and was associated with a high level of enzymatic resistance to d4T triphosphate (median, 16-fold; range, 5- to 48-fold). The role of K65R in d4T resistance was confirmed in site-directed mutants generated in three different RT backgrounds. Phenotypic assays based on recombinant single-cycle replication or a whole-virus multiple replication cycle were unable to detect d4T resistance in d4T-selected mutants with K65R but detected cross-resistance to other nucleoside RT inhibitors. Four of the six viruses that had 215C/D mutations at baseline acquired the 215Y mutation alone or in association with K65R. Mutants having K65R and T215Y replicated less efficiently than viruses that had T215Y only, suggesting that selection of T215Y in patients treated with d4T may be favored. Our results demonstrate that K65R plays a role in d4T resistance and indicate that resistance pathways for d4T and AZT may not be identical. Biochemical analysis and improved replication assays are both required for a full phenotypic characterization of resistance to d4T. These findings highlight the complexity of the genetic pathways of d4T resistance and its phenotypic expression. PMID- 12719560 TI - Redox-triggered infection by disulfide-shackled human immunodeficiency virus type 1 pseudovirions. AB - We previously described a human immunodeficiency virus type 1 (HIV-1) envelope mutant that introduces a disulfide bridge between the gp120 surface proteins and gp41 transmembrane proteins (J. M. Binley, R. W. Sanders, B. Clas, N. Schuelke, A. Master, Y. Guo, F. Kajumo, D. J. Anselma, P. J. Maddon, W. C. Olson, and J. P. Moore, J. Virol. 74:627-643, 2000). Here we produced pseudovirions bearing the mutant envelope and a reporter gene to examine the mutant's infectious properties. These pseudovirions attach to cells expressing CD4 and coreceptor but infect only when triggered with reducing agent, implying that gp120-gp41 dissociation is necessary for infection. Further studies suggested that virus entry was arrested after CD4 and coreceptor engagement. By measuring the activities of various entry inhibitors against the arrested intermediate, we found that gp120-targeting inhibitors typically act prior to virus attachment, whereas gp41 inhibitors are able to act postattachment. Unexpectedly, a significant fraction of antibodies in HIV-1-positive sera neutralized virus postattachment, suggesting that downstream fusion events and structures figure prominently in the host immune response. Overall, this disulfide-shackled virus is a unique tool with potential utility in vaccine design, drug discovery, and elucidation of the HIV-1 entry process. PMID- 12719562 TI - In vitro and in ovo expression of chicken gamma interferon by a defective RNA of avian coronavirus infectious bronchitis virus. AB - Coronavirus defective RNAs (D-RNAs) have been used for site-directed mutagenesis of coronavirus genomes and for expression of heterologous genes. D-RNA CD-61 derived from the avian coronavirus infectious bronchitis virus (IBV) was used as an RNA vector for the expression of chicken gamma interferon (chIFN-gamma). D RNAs expressing chIFN-gamma were shown to be capable of rescue, replication, and packaging into virions in a helper virus-dependent system following electroporation of in vitro-derived T7 RNA transcripts into IBV-infected cells. Secreted chIFN-gamma, under the control of an IBV transcription-associated sequence derived from gene 5 of the Beaudette strain, was expressed from two different positions within CD-61 and shown to be biologically active. In addition, following infection of 10-day-old chicken embryos with IBV containing D RNAs expressing chIFN-gamma, the allantoic fluid was shown to contain biologically active chIFN-gamma, demonstrating that IBV D-RNAs can express heterologous genes in vivo. PMID- 12719563 TI - Brome mosaic virus RNA syntheses in vitro and in barley protoplasts. AB - The RNA replicase extracted from Brome mosaic virus (BMV)-infected plants has been used to characterize the cis-acting elements for RNA synthesis and the mechanism of RNA synthesis. Minus-strand RNA synthesis in vitro requires a structure named stem-loop C (SLC) that contains a clamped adenine motif. In vitro, there are several specific requirements for SLC recognition. We examined whether these requirements also apply to BMV replication in barley protoplasts. BMV RNA3s with mutations in SLC were transfected into barley protoplasts, and the requirements for minus- and plus-strand replication were found to correlate well with the requirements in vitro. Furthermore, previous analysis of replicase recognition of the Cucumber mosaic virus (CMV) and BMV SLCs indicates that the requirements in the BMV SLC are highly specific. In protoplasts, we found that BMV RNA3s with their SLCs replaced with two different CMV SLCs were defective for replication. In vitro results generated with the BMV replicase and minimal-length RNAs generally agreed with those of in vivo BMV RNA replication. To extend this conclusion, we determined that, corresponding with the process of infection, the BMV replicases extracted from plants at different times after infection have different levels of recognition of the minimal promoters for plus- and minus strand RNA syntheses. PMID- 12719564 TI - Determination of infectious retrovirus concentration from colony-forming assay with quantitative analysis. AB - The colony formation assay is the most commonly used titration method for defining the concentration of replication-incompetent murine leukemia virus derived retroviral vectors. However, titer varies with target cell type and number, transduction time, and concentration of polycation (e.g., Polybrene). Moreover, because most of the viruses cannot encounter target cells due to Brownian motion, their short half-lives, and the requirement for target cell division for activity, the actual infectious retrovirus concentration in the collected supernatant is higher than the viral titer. Here we correlate the physical viral particle concentration with the infectious virus concentration and colony formation titer with the help of a mathematical model. Ecotropic murine leukemia retrovirus supernatant, collected from the GP+E86/LNCX retroviral vector producer cell line, was concentrated by centrifugation and further purified by a sucrose density gradient. The physical concentration of purified viral vectors was determined by direct particle counting with an electron microscope. The concentrations of fresh and concentrated supernatant were determined by a quantitative reverse transcriptase activity assay. Titration of all supernatants by neomycin-resistant colony formation assay was also performed. There were 767 +/- 517 physical viral particles per infectious CFU in the crude viral supernatant. However, the infectious viral concentration determined by mathematical simulation was 143 viral particles per infectious unit, which is more consistent with the concentration determined by particle counting in purified viral solution. Our results suggest that the mathematical model can be used to extract a more accurate and reliable concentration of infectious retrovirus. PMID- 12719567 TI - Vesicular stomatitis viruses with rearranged genomes have altered invasiveness and neuropathogenesis in mice. AB - Transcription of vesicular stomatitis virus is controlled by the position of a gene relative to the single 3' genomic promoter: promoter-proximal genes are transcribed at higher levels than those in more 5' distal positions. In previous work, we generated viruses having rearranged gene orders. These viruses had the promoter-proximal gene that encodes the nucleocapsid protein, N, moved to the second or fourth position in the genome in combination with the glycoprotein gene, G, moved from its usual promoter-distal fourth position to the first or third position. This resulted in three new viruses identified by the positions of the N and G genes in the gene order: G3N4, G1N4, and G1N2. The viruses G3N4 and G1N4 were attenuated for lethality in mice. In the present study, we addressed the basis of this attenuation by measuring the ability of each of the rearranged viruses to travel to and replicate in the olfactory bulb and brain following intranasal inoculation. In addition, the neuropathogenicity, serum cytokine levels, and immunoglobulin G isotype profiles in infected mice were determined. All the viruses reached the olfactory bulb and brain, but the outcomes of these infections were dramatically different. Viruses N1G4(wt) and G1N2 caused lethal encephalitis in 100% of animals within 7 days postinoculation; however, viruses G3N4 and G1N4 were cleared from the brain by 7 days postinoculation and all animals survived without apparent distress. The viruses differed in the distribution and intensity of lesions produced and the type and levels of cytokines induced. Animals inoculated with N1G4(wt) or G1N2 displayed extensive encephalitis and meningitis and had elevated levels of serum gamma interferon compared to what was seen with G3N4- or G1N4-infected mice. In contrast to what occurred with intranasal inoculation, all four viruses caused lethal encephalitis when administered by direct inoculation to the brain, a route that circumvents the majority of the host immune response, demonstrating that G3N4 and G1N4 were not deficient in their abilities to cause disease in the brain. These findings indicate that gene rearrangement and its consequent alteration of gene expression can, without any other changes, alter the viral spread and cytokine response following intranasal infection. PMID- 12719566 TI - Disruption of gammaherpesvirus 68 gene 50 demonstrates that Rta is essential for virus replication. AB - Gammaherpesvirus pathogenesis is dependent on the ability of these viruses to establish a lifelong latent infection and the ability to reactivate from latency. Immediate-early genes of theses viruses are thought to be critical regulators of lytic replication and reactivation from latency. The gene 50-encoded Rta is the only immediate-early gene product that appears to be conserved among all characterized gammaherpesviruses. Previous studies have demonstrated that, in Epstein-Barr virus (EBV), Kaposi's sarcoma-associated virus, and gammaherpesvirus 68 (gamma HV68, also referred to as murine gammaherpesvirus 68), ectopic expression of Rta in latently infected cell lines can lead to induction of the viral cycle. Recently, studies employing null mutants of EBV have provided a formal demonstration that both Rta and the BZLF1 gene product, Zta, the two EBV immediate-early gene products, are essential for EBV replication. Here we generate and characterize a gene 50-null mutant gamma HV68 and demonstrate that the gene 50 product Rta is essential for virus replication. Providing gamma HV68 Rta in trans was sufficient to restore replication of the gene 50-null virus. Notably, Rta expressed from the spliced form of the gene 50 transcript was sufficient to complement growth of the gene 50-null virus. In addition, we provide evidence that loss of Rta expression leads to a complete defect in viral DNA replication and a significant defect in late antigen expression. This work lays the foundation for characterizing the role of Rta in gamma HV68 chronic infection of mice. PMID- 12719568 TI - Direct relationship between suppression of virus-specific immunity and emergence of cytomegalovirus disease in simian AIDS. AB - Although opportunistic infections like cytomegalovirus (CMV) are common sequelae of end-stage AIDS, the immune events leading to CMV reactivation in human immunodeficiency virus (HIV)-infected individuals are not well defined. The role of cellular and humoral CMV-specific immune responses in immune control of latent CMV infection was evaluated prospectively in a cohort of 11 simian immunodeficiency virus (SIV)-infected CMV-seropositive rhesus macaques, 6 of whom had histologic evidence of CMV disease at death. Macaques with CMV disease differed from macaques without CMV disease in having significantly higher levels of plasma SIV RNA and CMV DNA and significantly lower titers of anti-CMV binding antibodies (Abs) at the time of death. A significant decline in anti-CMV Abs and CMV-specific CD4(+) and CD8(+) T lymphocytes over time was observed in the macaques with CMV disease, but not in the macaques without CMV disease. Reduction in CMV-specific CD8(+) T lymphocytes and anti-CMV neutralizing Abs was significantly correlated with a decline in CMV-specific CD4(+) T lymphocytes. Although declines in CMV-specific T lymphocytes alone were sufficient for reactivation of low-level CMV viremia, high-level viremia (>1,000 copies of CMV DNA per ml of plasma) was observed when anti-CMV neutralizing and binding Abs had also declined. Thus, the occurrence of CMV reactivation-associated disease in AIDS is associated with suppression of both cellular and humoral CMV-specific immune responses. The underlying mechanism may be a dysfunction of memory B and CD8(+) T lymphocytes associated with SIV-induced impairment of CMV-specific CD4(+) T-cell help. PMID- 12719569 TI - Human herpesvirus 8-encoded vGPCR activates nuclear factor of activated T cells and collaborates with human immunodeficiency virus type 1 Tat. AB - Human herpesvirus 8 (HHV-8), the etiologic agent of Kaposi's sarcoma (KS), encodes a chemokine receptor homologue, the viral G protein-coupled receptor (vGPCR), that has been implicated in KS pathogenesis. Expression of vGPCR constitutively activates several signaling pathways, including NF-kappa B, and induces the expression of proinflammatory and angiogenic factors, consistent with the inflammatory hyperproliferative nature of KS lesions. Here we show that vGPCR also constitutively activates the nuclear factor of activated T cells (NF-AT), another transcription factor important in regulation of the expression of inflammatory cytokines and related factors. NF-AT activation by vGPCR depended upon signaling through the phosphatidylinositol 3-kinase-Akt-glycogen synthetase kinase 3 (PI3-K/Akt/GSK-3) pathway and resulted in increased expression of NF-AT dependent cell surface molecules (CD25, CD29, Fas ligand), proinflammatory cytokines (interleukin-2 [IL-2], IL-4), and proangiogenic factors (granulocyte macrophage colony-stimulating factor GMCSF and TNF alpha). vGPCR expression also increased endothelial cell-T-cell adhesion. Although infection with HHV-8 is necessary to cause KS, coinfection with human immunodeficiency virus type 1 (HIV 1), in the absence of antiretroviral suppressive therapy, increases the risk of KS by many orders of magnitude. NF-AT and NF-kappa B activation by vGPCR was greatly increased by the HIV-1 Tat protein, although Tat alone had little effect on NF-AT. The enhancement of NF-AT by Tat appears to be mediated through collaborative stimulation of the PI3-K/Akt/GSK-3 pathway by vGPCR and Tat. Our data further support the idea that vGPCR contributes to the pathogenesis of KS by a paracrine mechanism and, in addition, provide the first evidence of collaboration between an HIV-1 protein and an HHV-8 protein. PMID- 12719570 TI - Comparison of adjuvant efficacy of herpes simplex virus type 1 recombinant viruses expressing TH1 and TH2 cytokine genes. AB - The adjuvant effects of cytokines in humoral and cell-mediated immunity to herpes simplex virus type 1 (HSV-1) have been examined in mice using HSV-1 recombinant viruses expressing murine interleukin-2 (IL-2), IL-4, or gamma interferon (IFN gamma) gene. Groups of naive BALB/c mice were immunized intraperitoneally with one or three doses of the HSV-1 recombinant viruses expressing IL-2, IL-4, or IFN gamma or with parental control virus. Despite similar replication kinetics, these three recombinant viruses elicited different immune responses to HSV-1 on immunization. Immunization with the recombinant virus expressing IL-4 elicited a humoral response of greater magnitude than immunization with the recombinant viruses expressing IL-2 or IFN-gamma or with parental virus. In contrast, immunization with recombinant virus expressing IL-2 elicited a higher cytotoxic T cell response than immunization with viruses expressing IL-4 or IFN-gamma. Stimulation in vitro of splenocytes obtained from the mice immunized with UV inactivated HSV-1 McKrae resulted in a T(H)1 pattern of cytokine expression irrespective of the recombinant virus used in the immunization. As observed for the parental virus, both CD4(+) and CD8(+) T cells contributed equally to the production of IL-2 by the splenocytes of mice immunized with any of the three recombinant viruses. However, the pattern of IFN-gamma production by CD4(+) and CD8(+) T cells differed according to the recombinant virus used. After lethal ocular challenge, all immunized mice were protected completely against death and manifestations of eye disease caused by HSV-1, which are typical responses in unimmunized mice. Mice immunized with IL-4-expressing virus cleared the virus from their eyes more rapidly than mice immunized with IL-2- or IFN-gamma expressing virus. Taken together, our results suggest that, in contrast to IFN gamma which did not exhibit an adjuvant effect, both IL-4 and IL-2 act as adjuvants in immunization with HSV, with IL-4 showing greater efficacy. PMID- 12719571 TI - Positive regulation of CXCR4 expression and signaling by interleukin-7 in CD4+ mature thymocytes correlates with their capacity to favor human immunodeficiency X4 virus replication. AB - The emergence of X4 human immunodeficiency virus type 1 (HIV-1) variants in infected individuals is associated with poor prognosis. One of the possible causes of this emergence might be the selection of X4 variants in some specific tissue compartment. We demonstrate that the thymic microenvironment favors the replication of X4 variants by positively modulating the expression and signaling of CXCR4 in mature CD4(+) CD8(-) CD3(+) thymocytes. Here, we show that the interaction of thymic epithelial cells (TEC) with these thymocytes in culture induces an upregulation of CXCR4 expression. The cytokine secreted by TEC, interleukin-7 (IL-7), increases cell surface expression of CXCR4 and efficiently overcomes the downregulation induced by SDF-1 alpha, also produced by TEC. IL-7 also potentiates CXCR4 signaling, leading to actin polymerization, a process necessary for virus entry. In contrast, in intermediate CD4(+) CD8(-) CD3(-) thymocytes, the other subpopulation known to allow virus replication, TEC or IL-7 has little or no effect on CXCR4 expression and signaling. CCR5 is expressed at similarly low levels in the two thymocyte subpopulations, and neither its expression nor its signaling was modified by the cytokines tested. This positive regulation of CXCR4 by IL-7 in mature CD4(+) thymocytes correlates with their high capacity to favor X4 virus replication compared with intermediate thymocytes or peripheral blood mononuclear cells. Indeed, we observed an enrichment of X4 viruses after replication in thymocytes initially infected with a mixture of X4 (NL4-3) and R5 (NLAD8) HIV strains and after the emergence of X4 variants from an R5 primary isolate during culture in mature thymocytes. PMID- 12719565 TI - Multiple viral genetic analyses detect low-level human immunodeficiency virus type 1 replication during effective highly active antiretroviral therapy. AB - To evaluate human immunodeficiency virus type 1 (HIV-1) replication and selection of drug-resistant viruses during seemingly effective highly active antiretroviral therapy (HAART), multiple HIV-1 env and pol sequences were analyzed and viral DNA levels were quantified from nucleoside analog-experienced children prior to and during a median of 5.1 (range, 1.8 to 6.4) years of HAART. Viral replication was detected at different rates, with apparently increasing sensitivity: 1 of 10 by phylogenetic analysis; 2 of 10 by viral evolution with increasing genetic distances from the most recent common ancestor (MRCA) of infection; 3 of 10 by selection of drug-resistant mutants; and 6 of 10 by maintenance of genetic distances from the MRCA. When four- or five-drug antiretroviral regimens were given to these children, persistent plasma viral rebound did not occur despite the accumulation of highly drug-resistant genotypes. Among the four children without genetic evidence of viral replication, a statistically significant decrease in the genetic distance to the MRCA was detected in three, indicating the persistence of a greater number of early compared to recent viruses, and their HIV-1 DNA decreased by > or =0.9 log(10), resulting in lower absolute DNA levels (P = 0.007). This study demonstrates the variable rates of viral replication when HAART has suppressed plasma HIV-1 RNA for years to a median of <50 copies/ml and that combinations of four or five antiretroviral drugs suppress viral replication even after short-term virologic failure of three-drug HAART and despite ongoing accumulation of drug-resistant mutants. Furthermore, the decrease of cellular HIV-1 DNA to low absolute levels in those without genetic evidence of viral replication suggests that monitoring viral DNA during HAART may gauge low level replication. PMID- 12719573 TI - Transduction of the mammary epithelium with adenovirus vectors in vivo. AB - Because the mammary parenchyma is accessible from the exterior of an animal through the mammary duct, adenovirus transduction holds promise for the short term delivery of genes to the mammary epithelium for both research and therapeutic purposes. To optimize the procedure and evaluate its efficacy, an adenovirus vector (human adenovirus type 5) encoding a green fluorescent protein (GFP) reporter and deleted of E1 and E3 was injected intraductally into the mouse mammary gland. We evaluated induction of inflammation (by intraductal injection of [(14)C]sucrose and histological examination), efficiency of transduction, and maintenance of normal function in transduced cells. We found that transduction of the total epithelium in the proximal portion of the third mammary gland varied from 7% to 25% at a dose of 2 x 10(6) PFU of adenovirus injected into day 17 pregnant mice. Transduction was maintained for at least 7 days with minimal inflammatory response; however, significant mastitis was observed 12 days after transduction. Adenovirus transduction could also be used in the virgin animal with little mastitis 3 days after transduction. Transduced mammary epithelial cells maintained normal morphology and function. Our results demonstrate that intraductal injection of adenovirus vectors provides a versatile and noninvasive method of investigating genes of interest in mouse mammary epithelial cells. PMID- 12719572 TI - Infection-triggered regulatory mechanisms override the role of STAT 4 in control of the immune response to influenza virus antigens. AB - Accurate control of the balance of the T1 and T2 cells during antiviral immunity is essential for optimizing immune effector functions and for avoiding potentially severe immunopathology. We examined the in vivo role of the signal transducer and activator of transcription (STAT) 4 in regulating the T1/T2 balance during the response to live influenza virus and isolated viral proteins. We found that the differentiation of gamma interferon (IFN-gamma)-producing Th1 and Tc1 cells after inoculation of live virus occurred independently of STAT 4 expression. Influenza virus-specific T2 and Tc2 responses were well controlled in such STAT 4-deficient mice unless IFN-gamma was eliminated as well. In contrast, the STAT 4-dependent signaling pathway played a more essential role in regulating the T1/T2 balance after immunization with viral proteins and, in particular, inactivated nonreplicating virus. Pulmonary infection was cleared even in the absence of both functional STAT 4 genes and functional IFN-gamma genes because virus-neutralizing antibodies were still generated, consistent with a substantial redundancy in different antiviral effector pathways. Thus, replicating agents such as live influenza virus can elicit IFN-gamma and control T2 immunity independently of STAT 4, whereas the profile of immunity to isolated proteins is more reliant on an intact STAT 4 signaling pathway. PMID- 12719574 TI - Comprehensive investigation of the molecular defect in vif-deficient human immunodeficiency virus type 1 virions. AB - Replication of human immunodeficiency virus type 1 (HIV-1) in primary blood lymphocytes, certain T-cell lines (nonpermissive cells), and most likely in vivo is highly dependent on the virally encoded Vif protein. Evidence suggests that Vif acts late in the viral life cycle during assembly, budding, and/or maturation to counteract the antiviral activity of the CEM15 protein and possibly other antiviral factors. Because HIV-1 virions produced in the absence of Vif are severely restricted at a postentry, preintegration step of infection, it is presumed that such virions differ from wild-type virions in some way. In the present study, we established a protocol for producing large quantities of vif deficient HIV-1 (HIV-1/Delta vif) from an acute infection of nonpermissive T cells and performed a thorough examination of the defect in these virions. Aside from the expected lack of Vif, we observed no apparent abnormalities in the packaging, modification, processing, or function of proteins in Delta vif virions. In addition, we found no consistent defect in the ability of Delta vif virions to perform intravirion reverse transcription under a variety of assay conditions, suggesting that the reverse transcription complexes in these particles can behave normally under cell-free conditions. Consistent with this finding, neither the placement of the primer tRNA3Lys nor its ability to promote reverse transcription in an in vitro assay was affected by a lack of Vif. Based on the inability of this comprehensive analysis to uncover molecular defects in Delta vif virions, we speculate that such defects are likely to be subtle and/or rare. PMID- 12719575 TI - Determination of minimum herpes simplex virus type 1 components necessary to localize transcriptionally active DNA to ND10. AB - DNA viruses such as herpes simplex virus type 1 (HSV-1) appear to start their replicative processes at specific nuclear domains known as ND10. In analyses to determine the minimum viral components needed for transcript accumulation at ND10, we find that a specific viral DNA sequence, OriS, and the viral immediate early proteins ICP4 and ICP27 are sufficient for a reporter gene placed in cis to the OriS sequence to transcribe at ND10. A chromatin immunoprecipitation assay demonstrated expected critical intermediates in retaining the minimal genome at ND10 for the HSV-1 replication origin through direct or indirect binding to the host protein Daxx. Coimmunoprecipitation assays with antibodies to Daxx and ICP4, ICP27, and ICP8 showed that the respective proteins interact, possibly forming a complex. A potential complex between the origin, early viral DNA-binding protein ICP8 and Daxx did not result in transcription at ND10. Thus, the deposition of transcriptionally active HSV-1 genomes at ND10 is most likely a consequence of retention at ND10 through the interaction of viral genome-bound ICP4 and ICP27 with Daxx. Such a complex might be more likely immobilized at the outside of ND10 by the PML-interacting Daxx than at other nuclear sites. PMID- 12719577 TI - Mutations proximal to the minor groove-binding track of human immunodeficiency virus type 1 reverse transcriptase differentially affect utilization of RNA versus DNA as template. AB - Human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT), like all retroviral RTs, is a versatile DNA polymerase that can copy both RNA and DNA templates. In spite of extensive investigations into the structure-function of this enzyme, the structural basis for this dual template specificity is poorly understood. Biochemical studies with two mutations in HIV-1 RT that affect residues contacting the template-primer now provide some insight into this specialized property. The mutations are N255D and N265D, both adjoining the minor groove-binding track, in the thumb region. The N265D substitution led to a loss of processive polymerization on DNA but not on RNA, whereas N255D drastically reduced processive synthesis on both templates. This differential template usage was accompanied by a rapid dissociation of the N265D variant on DNA but not RNA templates, whereas the N255D variant rapidly dissociated from both templates. Molecular dynamics modeling suggested that N265D leads to a loss of template strand-specific hydrogen bonding, indicating that this is a key determinant of the differential template affinity. The N255D substitution caused local changes in conformation and a consequent loss of interaction with the primer, leading to a loss of processive synthesis with both templates. We conclude that N265 is part of a subset of template-enzyme contacts that enable RT to utilize DNA templates in addition to RNA templates and that such residues play an important role in facilitating processive DNA synthesis on both RNA and DNA templates. PMID- 12719576 TI - Human immunodeficiency virus type 1 Env with an intersubunit disulfide bond engages coreceptors but requires bond reduction after engagement to induce fusion. AB - A mutant human immunodeficiency virus (HIV) envelope protein (Env) with an engineered disulfide bond between the gp120 and gp41 subunits (SOS-Env) was expressed on cell surfaces. With the disulfide bond intact, these cells did not fuse to target cells expressing CD4 and CCR5, but the fusion process did advance to an intermediate state: cleaving the disulfide bond with a reducing agent after but not before binding to target cells allowed fusion to occur. Through the use of an antibody directed against CCR5, it was found that at the intermediate stage, SOS-Env had associated with coreceptors. Reducing the disulfide bond after this intermediate had been reached resulted in hemifusion at low temperature and fusion at physiological temperature. The addition of C34 or N36, peptides that prevent six-helix bundle formation, at the hemifused state blocked the fusion that would have resulted after raising the temperature. Thus, Env has not yet folded into six-helix bundles after hemifusion has been achieved. Because SOS-Env binds CCR5, it is suggested that the conformational changes in wild-type Env that result from this binding cause disengagement of gp120 from gp41 in the region of the engineered bond. It is proposed that this disengagement is the event that directly frees gp41 to undergo the conformational changes that lead to fusion. The intermediate state achieved prior to reduction of the disulfide bond was stable. The capture of this configuration of Env could yield a suitable antigen for vaccine development, and it may also be a target for pharmacological intervention against HIV-1 entry. PMID- 12719578 TI - In vivo evolution of human immunodeficiency virus type 1 toward increased pathogenicity through CXCR4-mediated killing of uninfected CD4 T cells. AB - The destruction of the immune system by progressive loss of CD4 T cells is the hallmark of AIDS. CCR5-dependent (R5) human immunodeficiency virus type 1 (HIV-1) isolates predominate in the early, asymptomatic stages of HIV-1 infection, while CXCR4-dependent (X4) isolates typically emerge at later stages, frequently coinciding with a rapid decline in CD4 T cells. Lymphocyte killing in vivo primarily occurs through apoptosis, but the importance of apoptosis of HIV-1 infected cells relative to apoptosis of uninfected bystander cells is controversial. Here we show that in human lymphoid tissues ex vivo, apoptosis of uninfected bystander CD4 T cells is a major mechanism of lymphocyte depletion caused by X4 HIV-1 strains but is only a minor mechanism of depletion by R5 strains. Further, X4 HIV-1-induced bystander apoptosis requires the interaction of the viral envelope glycoprotein gp120 with the CXCR4 coreceptor on CD4 T cells. These results emphasize the contribution of bystander apoptosis to HIV-1 cytotoxicity and suggest that in association with a coreceptor switch in HIV disease, T-cell killing evolves from an infection-restricted stage to generalized toxicity that involves a high degree of bystander apoptosis. PMID- 12719579 TI - Reticuloendotheliosis virus sequences within the genomes of field strains of fowlpox virus display variability. AB - Nine field strains of fowlpox virus (FPV) isolated during a 24-year span from geographically diverse outbreaks of fowlpox in the United States were screened for the presence of reticuloendotheliosis virus (REV) sequences in their genomes by PCR. Each isolate appeared to be heterogeneous in that either a nearly intact provirus or just a 248- or 508-nucleotide fusion of portions of the integrated REV 5' and 3' long terminal repeats (LTRs) was exclusively present at the same genomic site. In contrast, four fowlpox vaccines of FPV origin and three originating from pigeonpox virus were genetically homogeneous in having retained only the 248-bp LTR fusion, whereas two other FPV-based vaccines had only the larger one. These remnants of integrated REV presumably arose during homologous recombination at one of the two regions common to both LTRs or during retroviral excision from the FPV genome. Loss of the provirus appeared to be a natural event because the tripartite population could be detected in a field sample (tracheal lesion). Moreover, the provirus was also readily deleted during propagation of FPV in cultured cells, as evidenced by the detection of truncated LTRs after one passage of a plaque-purified FPV recombinant having a "genetically marked" provirus. However, the deletion mutants did not appear to have a substantial replicative advantage in vitro because even after 55 serial passages the original recombinant FPV was still prevalent. As to the in vivo environment, retention of the REV provirus may confer some benefit to FPV for infection of poultry previously vaccinated against fowlpox. PMID- 12719581 TI - Role of the serine-threonine kinase PAK-1 in myxoma virus replication. AB - Subversion or appropriation of cellular signal transduction pathways is a common strategy employed by viruses to promote an environment within infected cells that supports the viral replicative cycle. Using subsets of 3T3 murine fibroblasts previously shown to differ in their ability to support myxoma virus (MV) replication, we investigated the role of host serine-threonine kinases (STKs) as potential mediators of the permissive phenotype. Both permissive and nonpermissive 3T3 cells supported equivalent levels of virion binding, entry, and early virus gene expression, indicating that MV tropism in 3T3 cells was not determined by receptor-mediated entry. In contrast, late virus gene expression and viral DNA replication were selectively compromised in restrictive 3T3 cells. Addition of specific protein kinase inhibitors, many of which shared the ability to influence the activity of the STKs p21-activated kinase 1 (PAK-1) and Raf-1 attenuated MV replication in permissive 3T3 cells. Western blot detection of the phosphorylated forms of PAK-1 (Thr423) and Raf-1 (Ser338) confirmed activation of these kinases in permissive cells after MV infection or gamma interferon treatment, but the activated forms of both kinases were greatly reduced or absent in restrictive 3T3 cells. The biological significance of these activations was demonstrated by using the autoinhibitory domain of PAK-1 (amino acids 83 to 149), expression of which reduced the efficiency of MV infection in permissive 3T3 cells concurrent with a decrease in PAK-1 activation. In comparison, overexpression of a constitutively active PAK-1 (T423E) mutant increased MV replication in restrictive 3T3 cells. These observations suggest that induced signaling via cellular STKs may play important roles in determining the permissiveness of host cells to poxvirus infection. PMID- 12719580 TI - Molecular features of the broadly neutralizing immunoglobulin G1 b12 required for recognition of human immunodeficiency virus type 1 gp120. AB - IgG1 b12 is a broadly neutralizing antibody against human immunodeficiency virus type 1 (HIV-1). The epitope recognized by b12 overlaps the CD4 receptor-binding site (CD4bs) on gp120 and has been a target for vaccine design. Determination of the three-dimensional structure of immunoglobulin G1 (IgG1) b12 allowed modeling of the b12-gp120 interaction in which the protruding third complementarity determining region (CDR) of the heavy chain (H3) was crucial for antibody binding. In the present study, extensive mutational analysis of the antigen binding site of Fab b12 was carried out to investigate the validity of the model and to identify residues important for gp120 recognition and, by inference, key to the anti-HIV-1 activity of IgG1 b12. In all, 50 mutations were tested: 40 in H3, 4 each in H2 and L1, and 2 in L3. The results suggest that the interaction of gp120 with H3 of b12 is crucially dependent not only on a Trp residue at the apex of the H3 loop but also on a number of residues at the base of the loop. The arrangement of these residues, including aromatic side chains and side chains that hydrogen bond across the base of the loop, may rigidify H3 for penetration of the recessed CD4-binding cavity. The results further emphasize the importance to gp120 binding of a Tyr residue at the apex of the H2 loop that forms a second finger-like structure and a number of Arg residues in L1 that form a positively charged, shelf-like structure. In general, the data are consistent with the b12 gp120 interaction model previously proposed. At the gene level, somatic mutation is seen to be crucial for the generation of many of the structural features described. The Fab b12 mutants were also tested against the b12 epitope-mimic peptide B2.1, and the reactivity profile had many similarities but also significant differences from that observed for gp120. The paratope map of b12 may facilitate the design of molecules that are able to elicit b12-like activities. PMID- 12719582 TI - Hyperglycosylated mutants of human immunodeficiency virus (HIV) type 1 monomeric gp120 as novel antigens for HIV vaccine design. AB - The ability to induce broadly neutralizing antibodies should be a key component of any forthcoming vaccine against human immunodeficiency virus type 1. One potential vaccine candidate, monomeric gp120, has generally failed to elicit such antibodies. We postulated that gp120 might be a better immunogen if it could be engineered to preferentially bind known broadly neutralizing antibodies. In a first study, we found that four alanine substitutions on the perimeter of the so called Phe-43 cavity of gp120 could reduce binding of weakly neutralizing CD4 binding site antibodies (R. Pantophlet, E. O. Saphire, P. Poignard, P. W. H. I. Parren, I. A. Wilson, and D. R. Burton, J. Virol. 77:642-658, 2003), while slightly enhancing binding of the potent, broadly neutralizing antibody b12. In the present study, we sought to reduce or abolish the binding of a wider range of nonneutralizing antibodies, by incorporating extra N-glycosylation motifs at select positions into the hypervariable loops and the gp120 core. A hyperglycosylated mutant containing seven extra glycosylation sequons (consensus sequences) and the four alanine substitutions described above did not bind an extensive panel of nonneutralizing and weakly neutralizing antibodies, including a polyclonal immunoglobulin preparation (HIVIG) of low neutralizing potency. Binding of b12, at lowered affinity, and of four antibodies to the C1 and C5 regions was maintained. Removal of N- and C-terminal residues in the C1 and C5 regions, respectively, reduced or abolished binding of the four antibodies, but this also adversely affected b12 binding. The hyperglycosylated mutant and its analogues described here are novel antigens that may provide a new approach to eliciting antibodies with b12-like neutralizing properties. PMID- 12719583 TI - Lentivirus vectors pseudotyped with filoviral envelope glycoproteins transduce airway epithelia from the apical surface independently of folate receptor alpha. AB - The practical application of gene therapy as a treatment for cystic fibrosis is limited by poor gene transfer efficiency with vectors applied to the apical surface of airway epithelia. Recently, folate receptor alpha (FR alpha), a glycosylphosphatidylinositol-linked surface protein, was reported to be a cellular receptor for the filoviruses. We found that polarized human airway epithelia expressed abundant FR alpha on their apical surface. In an attempt to target these apical receptors, we pseudotyped feline immunodeficiency virus (FIV) based vectors by using envelope glycoproteins (GPs) from the filoviruses Marburg virus and Ebola virus. Importantly, primary cultures of well-differentiated human airway epithelia were transduced when filovirus GP-pseudotyped FIV was applied to the apical surface. Furthermore, by deleting a heavily O-glycosylated extracellular domain of the Ebola GP, we improved the titer of concentrated vector severalfold. To investigate the folate receptor dependence of gene transfer with the filovirus pseudotypes, we compared gene transfer efficiency in immortalized airway epithelium cell lines and primary cultures. By utilizing phosphatidylinositol-specific phospholipase C (PI-PLC) treatment and FR alpha blocking antibodies, we demonstrated FR alpha-dependent and -independent entry by filovirus glycoprotein-pseudotyped FIV-based vectors in airway epithelia. Of particular interest, entry independent of FR alpha was observed in primary cultures of human airway epithelia. Understanding viral vector binding and entry pathways is fundamental for developing cystic fibrosis gene therapy applications. PMID- 12719584 TI - The latency-associated nuclear antigen homolog of herpesvirus saimiri inhibits lytic virus replication. AB - Herpesvirus saimiri (HVS), a T-lymphotropic tumor virus of neotropical primates, and the Kaposi's sarcoma-associated human herpesvirus 8 (KSHV) belong to the gamma-(2)-herpesvirus (Rhadinovirus) subfamily and share numerous features of genome structure and organization. The KSHV latency-associated nuclear antigen (LANA) protein appears to be relevant for viral persistence, latency, and transformation. It binds to DNA, colocalizes with viral episomal DNA, and presumably mediates efficient persistence of viral genomes. LANA further represses the transcriptional and proapoptotic activities of the p53 tumor suppressor protein. Here we report on the ORF73 gene of HVS strain C488, which is the positional and structural homolog of KSHV LANA. The ORF73 gene in OMK cells can encode a 62-kDa protein that localizes to the nucleus in a pattern similar to that of LANA. We show that the ORF73 gene product can regulate viral gene expression by acting as a transcriptional modulator of latent and lytic viral promoters. To define the HVS ORF73 function in the background of a replication competent virus, we constructed a viral mutant that expresses ORF73 under the transcriptional control of a mifepristone (RU-486)-inducible promoter. The HVS ORF73 gene product efficiently suppresses lytic viral replication in permissive cells, indicating that it defines a critical control point between viral persistence and lytic replication. PMID- 12719585 TI - Sodium-dependent myo-inositol transporter 1 is a cellular receptor for Mus cervicolor M813 murine leukemia virus. AB - Retrovirus infection is initiated by binding of the surface (SU) portion of the viral envelope glycoprotein (Env) to specific receptors on cells. This binding triggers conformational changes in the transmembrane portion of Env, leading to membrane fusion and cell entry, and is thus a major determinant of retrovirus tissue and species tropism. The M813 murine leukemia virus (MuLV) is a highly fusogenic gammaretrovirus, isolated from Mus cervicolor, whose host range is limited to mouse cells. To delineate the molecular mechanisms of its restricted host range and its high fusogenic potential, we initiated studies to characterize the cell surface protein that mediates M813 infection. Screening of the T31 mouse hamster radiation hybrid panel for M813 infectivity localized the receptor gene to the distal end of mouse chromosome 16. Expression of one of the likely candidate genes (slc5a3) within this region in human cells conferred susceptibility to both M813 infection and M813-induced fusogenicity. slc5a3 encodes sodium myo-inositol transporter 1 (SMIT1), thus adding another sodium dependent transporter to the growing list of proteins used by MuLVs for cell entry. Characterization of SMIT1 orthologues in different species identified several amino acid variations within two extracellular loops that may restrict susceptibility to M813 infection. PMID- 12719586 TI - Ribavirin treatment up-regulates antiviral gene expression via the interferon stimulated response element in respiratory syncytial virus-infected epithelial cells. AB - Respiratory syncytial virus (RSV) is a mucosa-restricted virus that is a leading cause of epidemic respiratory tract infections in children. RSV replication is a potent activator of the epithelial-cell genomic response, influencing the expression of a spectrum of cellular pathways, including proinflammatory chemokines of the CC, CXC, and CX(3)C subclasses. Ribavirin (1-beta-D ribofuranosyl-1,2,4-triazole-3-carboxamide) is a nontoxic antiviral agent currently licensed for the treatment of severe RSV lower respiratory tract infections. Because ribavirin treatment reduces the cytopathic effect in infected cells, we used high-density microarrays to investigate the hypothesis that ribavirin modifies the virus-induced epithelial genomic response to replicating virus. Ribavirin treatment administered in concentrations of 10 to 100 micro g/ml potently inhibited RSV transcription, thereby reducing the level of RSV N transcripts to approximately 13% of levels in nontreated cells. We observed that in both the absence and the presence of ribavirin, RSV infection induced global alterations in the host epithelial cell, affecting approximately 49% of the approximately 6,650 expressed genes detectable by the microarray. Ribavirin influences the expression of only 7.5% of the RSV-inducible genes (total number of genes, 272), suggesting that the epithelial-cell genetic program initiated by viral infection is independent of high-level RSV replication. Hierarchical clustering of the ribavirin-regulated genes identified four expression patterns. In one group, ribavirin inhibited the expression of the RSV-inducible CC chemokines MIP-1 alpha and -1 beta, which are important in RSV-induced pulmonary pathology, and interferon (IFN), a cytokine important in the mucosal immune response. In a second group, ribavirin further up-regulated a set of RSV- and IFN stimulated response genes (ISGs) encoding antiviral proteins (MxA and p56), complement products, acute-phase response factors, and the STAT and IRF transcription factors. Because IFN-beta expression itself was reduced in the ribavirin-treated cells, we further investigated the mechanism for up-regulation of the IFN-signaling pathway. Enhanced expression of IFI 6-16, IFI 9-27, MxA/p78, STAT-1 alpha, STAT-1 beta, IRF-7B, and TAP-1-LMP2 transcripts were independently reproduced by Northern blot analysis. Ribavirin-enhanced TAP-1-LMP2 expression was a transcriptional event where site mutations of the IFN-stimulated response element (ISRE) blocked RSV and ribavirin-inducible promoter activity. Furthermore, ribavirin up-regulated the transcriptional activity of a reporter gene selectively driven by the ISRE. In specific DNA pull-down assays, we observed that ribavirin enhanced RSV-induced STAT-1 binding to the ISRE. We conclude that ribavirin potentiates virus-induced ISRE signaling to enhance the expression of antiviral ISGs, suggesting a mechanism for the efficacy of combined treatment with ribavirin and IFN in other chronic viral diseases. PMID- 12719587 TI - Reovirus sigma NS and mu NS proteins form cytoplasmic inclusion structures in the absence of viral infection. AB - Reovirus replication occurs in the cytoplasm of infected cells and culminates in the formation of crystalline arrays of progeny virions within viral inclusions. Two viral nonstructural proteins, sigma NS and micro NS, and structural protein sigma 3 form protein-RNA complexes early in reovirus infection. To better understand the minimal requirements of viral inclusion formation, we expressed sigma NS, mu NS, and sigma 3 alone and in combination in the absence of viral infection. In contrast to its concentration in inclusion structures during reovirus replication, sigma NS expressed in cells in the absence of infection is distributed diffusely throughout the cytoplasm and does not form structures that resemble viral inclusions. Expressed sigma NS is functional as it complements the defect in temperature-sensitive, sigma NS-mutant virus tsE320. In both transfected and infected cells, mu NS is found in punctate cytoplasmic structures and sigma 3 is distributed diffusely in the cytoplasm and the nucleus. The subcellular localization of mu NS and sigma 3 is not altered when the proteins are expressed together or with sigma NS. However, when expressed with micro NS, sigma NS colocalizes with mu NS to punctate structures similar in morphology to inclusion structures observed early in viral replication. During reovirus infection, both sigma NS and mu NS are detectable 4 h after adsorption and colocalize to punctate structures throughout the viral life cycle. In concordance with these results, sigma NS interacts with mu NS in a yeast two-hybrid assay and by coimmunoprecipitation analysis. These data suggest that sigma NS and mu NS are the minimal viral components required to form inclusions, which then recruit other reovirus proteins and RNA to initiate viral genome replication. PMID- 12719588 TI - Differentiation of varicella-zoster virus ORF47 protein kinase and IE62 protein binding domains and their contributions to replication in human skin xenografts in the SCID-hu mouse. AB - To investigate the role of the ORF47 protein kinase of varicella-zoster virus (VZV), we constructed VZV recombinants with targeted mutations in conserved motifs of ORF47 and a truncated ORF47 and characterized these mutants for replication, phosphorylation, and protein-protein interactions in vitro and for infectivity in human skin xenografts in the SCID-hu mouse model in vivo. Previous experiments showed that ROka47S, a null mutant that makes no ORF47 protein, did not replicate in skin in vivo (J. F. Moffat, L. Zerboni, M. H. Sommer, T. C. Heineman, J. I. Cohen, H. Kaneshima, and A. M. Arvin, Proc. Natl. Acad. Sci. USA 95:11969-11974, 1998). The construction of VZV recombinants with targeted ORF47 mutations made it possible to assess the effects on VZV infection of human skin xenografts of selectively abolishing ORF47 protein kinase activity. ORF47 mutations that resulted in a C-terminal truncation or disrupted the DYS kinase motif eliminated ORF47 kinase activity and were associated with extensive nuclear retention of ORF47 and IE62 proteins in vitro. Disrupting ORF47 kinase function also resulted in a marked decrease in VZV replication and cutaneous lesion formation in skin xenografts in vivo. However, infectivity in vivo was not blocked completely as long as the capacity of ORF47 protein to bind IE62 protein was preserved, a function that we identified and mapped to the N-terminal domain of ORF47 protein. These experiments indicate that ORF47 kinase activity is of critical importance for VZV infection and cell-cell spread in human skin in vivo but suggest that it is the formation of complexes between ORF47 and IE62 proteins, both VZV tegument components, that constitutes the essential contribution of ORF47 protein to VZV replication in vivo. PMID- 12719589 TI - Kaposi's sarcoma-associated herpesvirus latency-associated nuclear antigen induces expression of the helix-loop-helix protein Id-1 in human endothelial cells. AB - Kaposi's sarcoma (KS)-associated herpesvirus (KSHV) (also known as human herpesvirus 8) is a gamma-2 herpesvirus believed to be the etiologic agent responsible for KS. The pathogenesis of this potentially life-threatening neoplasm is complex and unclear, and it is currently unknown how KSHV causes KS. Id (named for inhibitor of DNA binding or inhibitor of differentiation) proteins were identified in 1990 and found to be naturally occurring dominant-negative inhibitors of basic helix-loop-helix transcription factors. Id-1, the most well studied member of this family, has since been shown to play a key role in several biological systems including cellular differentiation, cell cycle regulation, and tumorigenesis. In this report, we demonstrate that Id-1 is expressed at high levels in KS tumor cells both in vitro and in vivo but is expressed at relatively modest levels in endothelial cells (ECs), the likely precursor of the KS tumor cell. Infection of precursor cells with KSHV may be responsible for this enhanced expression, as KSHV infection induced Id-1 27-fold in ECs under our experimental conditions. Furthermore, we demonstrate that the KSHV-encoded latency-associated nuclear antigen (LANA) protein appears to be involved. Expression of LANA in ECs resulted in Id-1 induction that was almost identical to the induction seen with KSHV-infected ECs. These results demonstrate the expression of Id-1 in KS tumor cells and indicate the KSHV LANA protein may be, at least in part, responsible. This may be an important mechanism by which KSHV allows KS tumor cells to escape normal cell cycle regulation and enhances their proliferation. PMID- 12719590 TI - The RNA subunit of telomerase is encoded by Marek's disease virus. AB - Marek's disease virus (MDV) is a herpesvirus of chickens that induces T lymphomas and tumors within 4 to 5 weeks of infection. Although the ability of MDV to induce tumors was demonstrated many years ago and although a number of viral oncogenic proteins have been identified, the mechanism by which the MDV is implicated in tumorigenesis is still unknown. We report the identification of a virus-encoded RNA telomerase subunit (vTR) within the genome of MDV. This gene is found in the genomic DNA of the oncogenic MDV strains, whereas it is not carried by the nononcogenic MDV strains. The vTR sequence exhibits 88% sequence identity with the chicken gene (cTR). Our functional analysis suggests that this telomerase RNA can reconstitute telomerase activity in a heterologous system (the knockout murine TR(-/-) cell line) by interacting with the telomerase protein component encoded by the host cell. We have also demonstrated that the vTR promoter region is efficient whatever the species of cell line considered and that vTR is expressed in vivo in peripheral blood leukocytes from chickens infected with the oncogenic MDV-RB1B and the vaccine MDV-Rispens strains. The functionality of the vTR gene and the potential implication of vTR in the oncogenesis induced by MDV is discussed. PMID- 12719591 TI - Reverse genetics for crimean-congo hemorrhagic fever virus. AB - The widespread geographical distribution of Crimean-Congo hemorrhagic fever (CCHF) virus (more than 30 countries) and its ability to produce severe human disease with high mortality rates (up to 60%) make CCHF a major public health concern worldwide. We describe here the successful establishment of a reverse genetics technology for CCHF virus, a member of the genus Nairovirus, family BUNYAVIRIDAE: The RNA polymerase I (pol I) system was used to generate artificial viral RNA genome segments (minigenomes), which contained different reporter genes in antisense (virus RNA) or sense (virus-complementary RNA) orientation flanked by the noncoding regions of the CCHF virus S segment. Reporter gene expression was observed in different eukaryotic cell lines following transfection and subsequent superinfection with CCHF virus, confirming encapsidation, transcription, and replication of the pol I-derived minigenomes. The successful transfer of reporter gene activity to fresh cells demonstrated the generation of recombinant CCHF viruses, thereby confirming the packaging of the pol I-derived minigenomes into progeny viruses. The system offers a unique opportunity to study the biology of nairoviruses and to develop therapeutic and prophylactic measures against CCHF infections. In addition, we demonstrated for the first time that the human pol I system can be used to develop reverse genetics approaches for viruses in the family BUNYAVIRIDAE: This is important since it might facilitate the manipulation of bunyaviruses with cell and host tropisms restricted to primates. PMID- 12719593 TI - The vaccinia virus I3L gene product is localized to a complex endoplasmic reticulum-associated structure that contains the viral parental DNA. AB - The vaccinia virus (VV) I3L gene product is a single-stranded DNA-binding protein made early in infection that localizes to the cytoplasmic sites of viral DNA replication (S. C. Rochester and P. Traktman, J. Virol. 72:2917-2926, 1998). Surprisingly, when replication was blocked, the protein localized to distinct cytoplasmic spots (A. Domi and G. Beaud, J. Gen. Virol. 81:1231-1235, 2000). Here these I3L-positive spots were characterized in more detail. By using an anti-I3L peptide antibody we confirmed that the protein localized to the cytoplasmic sites of viral DNA replication by both immunofluorescence and electron microscopy (EM). Before replication had started or when replication was inhibited with hydroxyurea or cytosine arabinoside, I3L localized to distinct cytoplasmic punctate structures of homogeneous size. We show that these structures are not incoming cores or cytoplasmic sites of VV early mRNA accumulation. Instead, morphological and quantitative data indicate that they are specialized sites where the parental DNA accumulates after its release from incoming viral cores. By EM, these sites appeared as complex, electron-dense structures that were intimately associated with the cellular endoplasmic reticulum (ER). By double labeling of cryosections we show that they contain DNA and a viral early protein, the gene product of E8R. Since E8R is a membrane protein that is able to bind to DNA, the localization of this protein to the I3L puncta suggests that they are composed of membranes. The results are discussed in relation to our previous data showing that the process of viral DNA replication also occurs in close association with the ER. PMID- 12719592 TI - Threonine 157 of influenza virus PA polymerase subunit modulates RNA replication in infectious viruses. AB - Previous results have shown a correlation between the decrease in protease activity of several influenza A virus PA protein mutants and the capacity to replicate of the corresponding mutant ribonucleoproteins (RNPs) reconstituted in vivo. In this work we studied the phenotype of mutant viruses containing these mutations. Viruses with a T162A mutation, which showed a very moderate decrease both in protease and replication activities of reconstituted RNPs, showed a wild type phenotype. Viruses with a T157A mutation, which presented a severe decrease in protease activity and replication of RNPs, showed a complex phenotype: (i) transport to the nucleus of PAT157A protein was delayed, (ii) virus multiplication was reduced at both low and high multiplicities, (iii) transcriptive synthesis was unaltered while replicative synthesis, especially cRNA, was diminished, and (iv) viral pathogenesis in mice was reduced, as measured by loss of body weight and virus titers in lungs. Finally, recombinant viruses with a T157E mutation in PA protein, which resulted in a drastic reduction of protease and replication activities of RNPs, were not viable. These results indicate that residue T157 in PA protein is important for the capacity of viral polymerase to synthesize cRNA. PMID- 12719594 TI - EBNA2 and activated Notch induce expression of BATF. AB - The immortalization of human B lymphocytes by Epstein-Barr virus (EBV) requires the virus-encoded transactivator EBNA2 and the products of both viral and cellular genes which serve as EBNA2 targets. In this study, we identified BATF as a cellular gene that is up-regulated dramatically within 24 h following the infection of established and primary human B cells with EBV. The transactivation of BATF is mediated by EBNA2 in a B-cell-specific manner and is duplicated in non EBV-infected B cells by the expression of mammalian Notch proteins. In contrast to other target genes activated by EBNA2, the BATF gene encodes a member of the AP-1 family of transcription factors that functions as a negative regulator of AP 1 activity and as an antagonist of cell growth. A potential role for BATF in promoting EBV latency is supported by studies in which BATF was shown to negatively impact the expression of a BZLF1 reporter gene and to reduce the frequency of lytic replication in latently infected cells. The identification of BATF as a cellular target of EBV provides important new information on how programs of viral and cellular gene expression may be coordinated to promote viral latency and control lytic-cycle entry. PMID- 12719595 TI - Discordant outcomes following failure of antiretroviral therapy are associated with substantial differences in human immunodeficiency virus-specific cellular immunity. AB - Many individuals chronically infected with human immunodeficiency virus type 1 (HIV-1) experience a recrudescence of plasma virus during continuous combination antiretroviral therapy (ART) due either to the emergence of drug-resistant viruses or to poor compliance. In most cases, virologic failure on ART is associated with a coincident decline in CD4(+) T lymphocyte levels. However, a proportion of discordant individuals retain a stable or even increasing CD4(+) T lymphocyte count despite virological failure. In order to address the nature of these different outcomes, we evaluated virologic and immunologic variables in a prospective, single-blinded, nonrandomized cohort of 53 subjects with chronic HIV 1 infection who had been treated with continuous ART and monitored intensively over a period of 19 months. In all individuals with detectable viremia on ART, multiple drug resistance mutations with similar impacts on viral growth kinetics were detected in the pol gene of circulating plasma virus. Further, C2V3 env gene analysis demonstrated sequences indicative of CCR5 coreceptor usage in the majority of those with detectable plasma viremia. In contrast to this homogeneous virologic pattern, comprehensive screening with a range of antigens derived from HIV-1 revealed substantial immunologic differences. Discordant subjects with stable CD4(+) T lymphocyte counts in the presence of recrudescent virus demonstrated potent virus-specific CD4(+) and CD8(+) T lymphocyte responses. In contrast, subjects with virologic failure associated with declining CD4(+) T lymphocyte counts had substantially weaker HIV-specific CD4(+) T lymphocyte responses and exhibited a trend towards weaker HIV-specific CD8(+) T lymphocyte responses. Importantly the CD4(+) response was sustained over periods as long as 11 months, confirming the stability of the phenomenon. These correlative data lead to the testable hypothesis that the consequences of viral recrudescence during continuous ART are modulated by the HIV-specific cellular immune response. PMID- 12719596 TI - The NB protein of influenza B virus is not necessary for virus replication in vitro. AB - The NB protein of influenza B virus is thought to function as an ion channel and therefore would be expected to have an essential function in viral replication. Because direct evidence for its absolute requirement in the viral life cycle is lacking, we generated NB knockout viruses by reverse genetics and tested their growth properties both in vitro and in vivo. Mutants not expressing NB replicated as efficiently as the wild-type virus in cell culture, whereas in mice they showed restricted growth compared with findings for the wild-type virus. Thus, the NB protein is not essential for influenza B virus replication in cell culture but promotes efficient growth in mice. PMID- 12719597 TI - Amphipathic helix-dependent localization of NS5A mediates hepatitis C virus RNA replication. AB - We identified an N-terminal amphipathic helix (AH) in one of hepatitis C virus (HCV)'s nonstructural proteins, NS5A. This AH is necessary and sufficient for membrane localization and is conserved across isolates. Genetically disrupting the AH impairs HCV replication. Moreover, an AH peptide-mimic inhibits the membrane association of NS5A in a dose-dependent manner. These results have exciting implications for the HCV life cycle and novel antiviral strategies. PMID- 12719598 TI - Construction of varicella-zoster virus recombinants from parent Oka cosmids and demonstration that ORF65 protein is dispensable for infection of human skin and T cells in the SCID-hu mouse model. AB - We generated an ORF65 deletion mutant by using a cosmid system constructed from the genome of a low-passage clinical isolate (P-Oka). Using the SCID-hu mouse model, we demonstrated that the ORF65 protein is dispensable for viral replication in skin and T cells, which are critical host cell targets during primary varicella-zoster virus infection. PMID- 12719599 TI - RNA interference of human papillomavirus type 18 E6 and E7 induces senescence in HeLa cells. AB - The human papillomavirus oncoproteins E6 and E7 promote cell proliferation and contribute to carcinogenesis by interfering with the activities of cellular tumor suppressors. We used a small interfering RNA molecule targeting the E7 region of the bicistronic E6 and E7 mRNA to induce RNA interference, thereby reducing expression of E6 and E7 in HeLa cells. RNA interference of E6 and E7 also inhibited cellular DNA synthesis and induced morphological and biochemical changes characteristic of cellular senescence. These results demonstrate that reducing E6 and E7 expression is sufficient to cause HeLa cells to become senescent. PMID- 12719600 TI - Green fluorescent protein-tagged retroviral envelope protein for analysis of virus-cell interactions. AB - Fluorescent retroviral envelope (Env) proteins were developed for direct visualization of viral particles. By fusing the enhanced green fluorescent protein (eGFP) to the N terminus of the amphotropic 4070A envelope protein, extracellular presentation of eGFP was achieved. Viruses incorporated the modified Env protein and efficiently infected cells. We used the GFP-tagged viruses for staining retrovirus receptor-positive cells, thereby circumventing indirect labeling techniques. By generating cells which conditionally expressed the GFP-tagged Env protein, we could confirm an inverse correlation between retroviral Env expression and infectivity (superinfection). eGFP-tagged virus particles are suitable for monitoring the dynamics of virus-cell interactions. PMID- 12719601 TI - Two major histocompatibility complex class I-restricted epitopes of the Borna disease virus p10 protein identified by cytotoxic T lymphocytes induced by DNA based immunization. AB - Borna disease virus (BDV) infection of Lewis rats is the most studied animal model of Borna disease, an often fatal encephalomyelitis. In this experimental model, BDV-specific CD8(+) cytotoxic T lymphocytes (CTLs) play a prominent role in the immunopathogenesis of infection by the noncytolytic, persistent BDV. Of the six open reading frames of BDV, CTLs to BDV X (p10) and the L-polymerase have never been studied. In this study, we used plasmid immunization to investigate the CTL response to BDV X and N. Plasmid-based immunization was a potent CTL inducer in Lewis rats. Anti-X CTLs were primed by a single injection of the p10 cDNA. Two codominant p10 epitopes, M(1)SSDLRLTLL(10) and T(8)LLELVRRL(16), associated with the RT1.A(l) major histocompatibility complex class I molecules of the Lewis rats, were identified. In addition, immunization with a BDV p40 expressing plasmid confirmed the previously reported RT1.A(l)-restricted A(230)SYAQMTTY(238) peptide as the CTL target for BDV N. In contrast to the CTL responses, plasmid vaccination was a poor inducer of an antibody response to p10. Three injections of a recombinant eukaryotic expression plasmid of BDV p10 were needed to generate a weak anti-p10 immunoglobulin M response. However, the antibody response could be optimized by a protein boost after priming with cDNA. PMID- 12719602 TI - In situ characterization of Cymbidium Ringspot Tombusvirus infection-induced posttranscriptional gene silencing in Nicotiana benthamiana. AB - In plants, posttranscriptional gene silencing (PTGS) is an ancient and effective defense mechanism against viral infection. A number of viruses encode proteins that suppress virus-activated PTGS. The p19 protein of tombusviruses is a potent PTGS suppressor which interferes with the onset of PTGS-generated systemic signaling and is not required for viral replication or for viral movement in Nicotiana benthamiana. This unique feature of p19 suppressor allowed us to analyze the mechanism of PTGS-based host defense and its viral suppression without interfering with other viral functions. In contrast to the necrotic symptoms caused by wild-type tombusvirus, the infection of p19-defective mutant virus results in the development of a typical PTGS-associated recovery phenotype in N. benthamiana. In this report we show the effect of PTGS on the viral infection process for N. benthamiana infected with either wild-type Cymbidium Ringspot Tombusvirus (CymRSV) or a p19-defective mutant (Cym19stop). In situ analyses of different virus-derived products revealed that PTGS is not able to reduce accumulation of virus in primary infected cells regardless of the presence of p19 PTGS suppressor. We also showed that both CymRSV and Cym19stop viruses move systemically in the vasculature, with similar efficiencies. However, in contrast to the uniform accumulation of CymRSV throughout systemically infected leaves, the presence of Cym19stop virus was confined to and around the vascular bundles. These results suggest that the role of p19 is to prevent the onset of mobile signal-induced systemic PTGS ahead of the viral infection front, leading to generalized infection. PMID- 12719603 TI - Induction of broad and potent anti-human immunodeficiency virus immune responses in rhesus macaques by priming with a DNA vaccine and boosting with protein adsorbed polylactide coglycolide microparticles. AB - Several vaccine technologies were evaluated for their abilities to induce anti human immunodeficiency virus Gag immune responses in rhesus macaques. While no vaccine alone was able to induce broad and strong immune responses, these were achieved by priming with Gag DNA and boosting with Gag protein adsorbed to polylactide coglycolide microparticles. This regimen elicited strong antibodies, helper T cells, and cytotoxic T lymphocytes and thus holds promise as an effective vaccination scheme. PMID- 12719605 TI - Interactions between Chlamydia pneumoniae and trace elements: a possible link to aortic valve sclerosis. AB - An association between Chlamydia pneumoniae and atherosclerotic cardiovascular diseases has been suggested. However, other factors may interact in the pathogenesis of valve sclerosis. Therefore, trace elements important for C. pneumoniae growth and host defense and markers of C. pneumoniae infection were studied in sclerotic valves and serum. Forty-six patients undergoing surgical valve replacement due to advanced aortic sclerosis were prospectively studied. Valves from 15 forensic cases with no heart valve disease and plasma from 46 healthy volunteers served as controls. C. pneumoniae was detected in 16/46 (34.8 %) sclerotic valves and in 0/15 forensic controls. IgG and IgA antibodies to C. pneumoniae were present in 54.3% and 26.1 % patients, respectively. In the patients' valves, iron, magnesium, and zinc each correlated to calcium, a marker of the histopathological severity of disease. Patients showed 10- to 70-fold increases of these trace elements in valves and an increased copper/zinc ratio in serum. In a majority of aortic sclerosis patients, one of several markers of C. pneumoniae infection were detected and all patients had a disturbed trace element balance in valves and serum suggestive of active immune process and infection. The pattern of trace element changes was essentially similar regardless of positive makers of C. pneumoniae, suggesting a similar etiopathogenesis in both subgroups. The 20-fold increase in iron, essential for C. pneumoniae growth, in sclerotic valves suggests a new possible link to this infection in aortic sclerosis. PMID- 12719606 TI - Sequential changes in Fe, Cu, and Zn in target organs during early Coxsackievirus B3 infection in mice. AB - In Coxsackievirus B3 (CB3) infection, the heart and pancreas are major target organs and, as a general host response, an associated immune activation and acute phase reaction develops. Although iron (Fe), copper (Cu), and zinc (Zn) are involved in these responses, sequential trace element changes in different target organs of infection have not been studied to date. In the present study, Fe, Cu, and Zn were measured through inductively coupled plasma mass spectrometry (ICP MS) in the plasma, liver, spleen, heart, and pancreas during the early phase (d 1 and 3) of CB3 infection in female Balb/c mice. The severity of the infection was assessed through clinical signs of disease and histopathology of the heart and pancreas, including staining of CD4 and CD8 cells in the pancreas. During infection, the concentrations of Fe, Cu, and Zn changed in the plasma, liver, and pancreas, but not in the spleen and heart. The changes in plasma Cu, Zn, and Fe seemed to be biphasic with a decrease at d 1 that turned into increased levels by d 3. Cu showed similar biphasic changes in the liver, spleen, and pancreas, whereas, for Zn and Fe, this pattern was only evident in the liver. In the pancreas, the reverse response occurred with pronounced decreases in Fe (23%, p < 0.05) and Zn (64%, p < 0.01) at d 3. Although the pathophysiological interpretation of these findings requires further research, the sequential determination of these elements may be of clinical value in enterovirus infections in deciding the stage of disease development. PMID- 12719607 TI - Iron depletion without anemia is not associated with impaired selenium status in college-aged women. AB - Iron-deficiency anemia has been shown to alter body mineral concentrations and activities of iron- and non-iron-containing enzymes, especially those with antioxidant functions. These effects, however, have been less studied in nonanemic iron-depleted individuals. Thus, this study assessed indices of selenium status in 12 college-aged females with adequate iron stores and 15 college-aged females with low iron stores before and after iron therapy. Blood samples were drawn at baseline for both groups and following iron supplementation in the low-iron-stores group. Hematocrit, hemoglobin, and serum ferritin concentrations of the low iron- stores group were significantly lower than those of the control group. The serum transferrin receptor-to-serum ferritin ratio in the low-iron stores group was significantly greater than that of the control group. Serum selenium and glutathione peroxidase concentrations of the low-iron stores group were not significantly different from those of the controls. Iron supplementation significantly increased hemoglobin, hematocrit, and serum ferritin concentrations and significantly decreased the serum transferrin receptor concentration and serum transferrin receptor:serum ferritin ratio in the low-iron-stores group posttreatment compared to pretreatment. Serum selenium and glutathione peroxidase concentrations did not differ significantly from pretreatment to posttreatment in the low-iron-stores group. Results of this study indicate that low iron stores without anemia are not associated with impaired selenium status in college-aged females. PMID- 12719608 TI - Serum copper levels and not zinc are positively associated with serum leptin concentrations in the healthy adult population. AB - Leptin, the obesity gene protein product, is a hormone with multiple physiological functions in the human. However, there are few reports in the literature on its role in trace element metabolism in the normal population. Therefore, we investigated the association among serum leptin, zinc, copper, and zinc/copper ratio in 570 healthy men and women aged 15 yr and older. Serum leptin assay was done with a commercial enzymelinked immunosorbent assay kit; serum zinc and copper levels were measured by an atomic absorption spectrophotometer. Serum leptin was found to be positively associated with age (r=0.254, p<0.001), sex (r=0.406, p<0.001), body mass index (BMI) (r=0.553, p<0.001), and serum copper (r=0.419, p<0.001), but negatively associated with the zinc/copper ratio (r= 0.423, p<0.001). There was no significant association between serum leptin and zinc (r= -0.131, p>0.05). When the confounding effects of age, sex, and BMI were removed, serum leptin was still positively associated with serum copper (r=0.197, p=0.02) and the serum zinc/copper ratio (r=-0.182, p=0.03). These results suggest that copper and not zinc has an effect on serum leptin levels. PMID- 12719609 TI - Selenium concentration in blood and hair of holstein dairy cows. AB - Four-hundred Holstein cows in 40 dairy farms in north Greece were included in this study, and blood (n=400), black hair (n=400), white hair (n=40), and feed (n=40) samples were obtained. Although the feeding regime in these farms was similar, the selenium content of feeds was variable. The Se content of concentrate feeds was 0.104+/-0.086 mg/kg dry matter (DM), and of silage, it was 0.025+/-0.018 mg/kg. A significantly positive correlation was found between the Se concentration in black hair and the Se concentration in blood (r2=0.610, p<0.001), the Se concentration in white hair and the Se concentration in blood (r2=0.770, p<0.001), and the Se concentration in white hair and the Se concentration in black hair (r2=0.921, p<0.001). The Se concentration in white hair was significantly smaller than that in black hair (p<0.001). PMID- 12719610 TI - Inhibition of glutathione reductase by cadmium ion in some rabbit tissues and the protective role of dietary selenium. AB - This study is aimed at investigating the inhibitory effect of cadmium ion on glutathione reductase activity of rabbit brain and liver and the relationship of this effect with dietary selenium. For this purpose, one group of New Zealand rabbits were fed a selenium-deficient diet, another group was fed a selenium-rich diet, and the control group was fed a normal diet. The brain and liver tissues of these groups were investigated for the in vitro inhibitory effects of Cd2+ on glutathione reductase activity. For liver, the percentage inhibition of glutathione reductase by 40 nmol/mg protein of Cd2+ was similar for selenium deficient and control groups, but significantly lower in the selenium-rich group. For brain tissues, there was no difference with respect to cadmium inhibition of glutathione reductase in all three groups. PMID- 12719611 TI - Dietary Zn deficiency does not influence systemic blood pressure and vascular nitric oxide signaling in normotensive rats. AB - Because zinc (Zn) is an important component for cell protection against certain oxygen species, it has been suggested that Zn deficiency impairs the potent oxidant defense capacity, which is constitutively provided in the vascular system. However, the influence of dietary Zn deficiency on systemic blood pressure and vascular system is controversial and unclear. We therefore examine the effect of dietary Zn deficiency on systemic blood pressure, a potent superoxide scavenger, aortic Cu/Zn superoxide dismutase (SOD) activity, a most representative synthase of the endothelium-derived relaxing factor, and aortic endothelial nitric oxide synthase (eNOS) expression. Furthermore, the direct effects of intravenous administration of NOS inhibitor, Nomega-nitro-L-arginine methyl ester (LNAME), and a SOD mimetic compound, tempol, in normotensives were tested in Wistar-Kyoto (WKY) rats. A Zn-deficient diet (4 wk) contributed to growth retardation, the decrease in thymus weight, and the lower levels of serum Zn compared with the standard diet group. However, no significant difference in conscious systolic and diastolic blood pressure was found in the Zn-deficiency group. The administration of L-NAME caused an increase in the mean arterial pressure (MAP) levels in the two groups of rats and the involvement of the vasodilator nitric oxide (NO) in the regulation of systemic BP in the normotensive state. On the other hand, administration of the superoxide scavenger, tempol, led to a decrease in MAP levels in the two groups of rats, indicating the participation of the oxygen free radical, superoxide, in the maintenance of the systemic BP in a normotensive state. There were no significant differences between the Zn-deficient diet group and the standard diet group in the normotensive state. eNOS expression and Cu/Zn SOD activity in the aorta were also intact in Zn-deficient normotensive rats. These findings suggest that the 4 wk of Zn deficiency was inadequate to alter systemic blood pressure and focal NO signaling in the normotensive state. Long-term Zn deficiency affects the neuronal, immune, and hematopoietic systems, which contribute to systemic and/or local circulation. However, Zn deficiency alone does not cause hypertension and local vascular dysfunction in the normotensive state. PMID- 12719612 TI - Effects of acute exercise on the levels of iron, magnesium, and uric acid in liver and spleen tissues. AB - In this study, we investigated the effects of acute exercise on tissue levels of iron, magnesium, and uric acid of rats. Twenty adult Wistar albino rats were used for the study. They were divided into two groups: controls (n=10) and the study group (n=10). The study group was left into a small water pool and allowed to do swimming exercise for 30 min while controls rested. All of the animals were sacrificed, and their livers and spleens removed and homogenized immediately. The iron, magnesium, and uric acid levels of the homogenates were measured by an autoanalyzer (ILAB 900, Italy) with commercial kits from the same company. Results were evaluated by the Mann-Whitney U-test. According to our results, the liver iron levels increased significantly with exercise, whereas spleen iron levels decreased significantly (p<0.05) compared to controls. We found no significant differences in the levels of the other two parameters with exercise. These results show that the iron distribution in organs changes with exercise. PMID- 12719613 TI - Mechanism of physiological effects of titanium leaf sprays on plants grown on soil. AB - Titanium (Ti) has significant biological effects on plants, being beneficial at low and toxic at higher concentrations. From results of our hydroponical experiment with oats, we have recently proposed that the effect called hormesis is the mechanism of Ti action in plants. Here, we present the experiment with oats (Avena sativa L. cv. Zlat'ak) grown on soil where Ti was applied using leaf sprays. Two different soils, three different concentrations of Ti(IV) citrate spray solution (0, 20, and 50 mg Ti/kg), and three different Mg concentrations in each soil were tested. Some physiological parameters (dry and raw weights, top heights, chlorophyll content) and element contents (Mg, Fe, Zn, Mn) were determined. Ti showed considerable effects on all physiological parameters and the element's contents were determined. Differences between the two different soil types used was only in the strength of the effect of Ti; the trends remained unchanged. Generally, the effect of Ti is considerably weaker if Ti is applied on leaves than if being added to the nutrient solution. Thus, we confirm here that the action of Ti on plants could be explained by the hormesis effect. PMID- 12719615 TI - Advances in medical science. Guest editorial. PMID- 12719614 TI - The strong correlation between serum copper level and the copper/zinc ratio to histopathological changes, clinical stage, and prognosis of Hodgkin's disease. PMID- 12719616 TI - Preliminary report of a NeuroPage service within a health care system. AB - NeuroPage, a paging service designed to reduce the everyday memory and/or planning problems of people with neurological deficits, is described. Following several research studies carried out to evaluate the system, a British local health authority set up a nationwide commercial NeuroPage service. A report on the first 40 clients recruited to the service is provided. The age range was 14 81 years, the majority of clients were men and the most frequent diagnosis was traumatic brain injury although a number of different diagnostic groups were represented. The types of messages sent and the group's satisfaction with NeuroPage are considered. Three brief case studies to illustrate the different ways clients used the system are presented. PMID- 12719617 TI - Prosthetic systems for addressing problems with initiation: guidelines for selection, training, and measuring efficacy. AB - In this paper we address the practical considerations for rehabilitation of problems with initiation. A variety of rehabilitation strategies for clients who have problems with initiation are discussed, as are our recommendations for the use of an external cueing system with these clients. Available external cueing systems are reviewed, including those that have been shown to be effective for prospective memory deficits. We identify important factors involved in selecting a particular external cueing device, and we provide guidelines for training to both respond to and program external cues. Finally, we discuss ways to monitor the efficacy of rehabilitation of problems with initiation. Throughout we provide an illustrative case example describing implementation of an external cueing system with a client who has difficulties with initiation, other aspects of executive function, and anterograde amnesia. PMID- 12719618 TI - Learning to live independently with expert systems in memory rehabilitation. AB - Expert systems (ES), which are a branch of artificial intelligence, has been widely used in different applications, including medical consultation and more recently in rehabilitation for assessment and intervention. The development and validation of an expert system for memory rehabilitation (ES-MR) is reported here. Through a web-based platform, ES-MR can provide experts with better decision making in providing intervention for persons with brain injuries, stroke, and dementia. The application and possible commercial production of a simultaneously developed version for "non-expert" users is proposed. This is especially useful for providing remote assistance to persons with permanent memory impairment when they reach a plateau of cognitive training and demand a prosthetic system to enhance memory for day-to-day independence. The potential use of ES-MR as a cognitive aid in conjunction with WAP mobile phones, Bluetooth technology, and Personal Digital Assistants (PDAs) is suggested as an avenue for future study. PMID- 12719619 TI - The essential steps cognitive orthotic. AB - A multifunction cognitive orthotic system is designed to assist people with cognitive disorders in the performance of ecologically relevant activities. This personal computer-based compensatory approach applies neuroscience and technology to the rehabilitation needs and challenges of people with chronic brain damage. The primary goal of this approach is the independent accomplishment of practical, applied tasks integral to daily living, at home, at work, at school and in the community. This paper will describe the "Essential Steps" cognitive orthotic system, and describe its potential to facilitate skill acquisition and enhance self-sufficient life management. The software was designed to guide and protect the user as he or she progresses through each "essential step" in the progression required for completion of a targeted task. The discussion will explicate the underlying principles promoting skill acquisition with this application of a cognitive orthotic. Data have shown that this compensatory aid can promote rapid skill acquisition and support reliable performance, even long past the time when rapid recovery and progress are customarily expected. Discussion will focus on issues of design for people with brain damage, the significance of this system's prototypical structure, and research findings and clinical observations made with people using "Essential Steps". PMID- 12719620 TI - What do brain dysfunctional patients report following memory compensation training? AB - Twenty five (25) out of 29 patients (86%) with unequivocal memorydisorders who received Memory Compensation Training (MCT) completed both a brief survey regarding their use of memory compensations and the Memory Compensation Questionnaire. Twenty-two (22) of the 25 patients (88%) reported daily use of memory compensation several months to years after the onset of their memory problems. The benefits included, by their report, being more productive, less disorganized, and less confused. Patients with memory disorders, who received MCT, also reported more frequent use of compensations than older normal adults studied by Dixon et al. [6]. The present sample of patients report proportionally spending more time to remember and greater reliance on those around them to help them to remember. The findings suggest that patients who are taught memory compensations tend to use them after the training period and show a different pattern of compensatory activities compared to normal older adults. PMID- 12719621 TI - Effectiveness of the ISAAC cognitive prosthetic system for improving rehabilitation outcomes with neurofunctional impairment. AB - Cognitive rehabilitation has the capacity to empower persons with brain-injuries and help them achieve heightened functional, personal, and social interactions within their environments. Interventions aimed at compensation for deficits and adaptation to cognitive disability can be aided through the use of assistive technology devices (ATD's). ATDs allow for their users to experience greater levels of independence, as well as social and vocational participation, which leads to a higher quality of life. The ISAAC system is a small, individualized, wearable cognitive prosthetic assistive technology system. Being fully individualized and very easy to use makes this system adaptable to, and appropriate for, patients with a wide variety of cognitive disabilities ranging from individuals with developmental disabilities to high functioning survivors of brain injury. The current article will discuss two cases that illustrate the effectiveness of the ISAAC system in assisting patients with generalization of rehabilitation to their home environments. Both patients incurred significant cognitive impairment, for which they were able to successfully compensate with the assistance of their ISAAC systems. These two case studies are typical examples of the functional independence that can be achieved through the use of the ISAAC system. When patients are properly selected for use of this system, appropriate content is authored, and sufficient training on the system is provided, the ISAAC system can prove very effective at improving patients' functional independence. PMID- 12719622 TI - Facilitation of skilled finger movements by repetitive peripheral magnetic stimulation (RPMS) - a new approach in central paresis. AB - A new therapeutic method for the rehabilitation of central paresis of the upper extremity, especially of fine skilled finger movements, is presented. The therapeutic concept is the activation of reorganization processes in the CNS. These processes are elicited by the induction of proprioceptive input to the CNS which corresponds physiologically to the lost input during active movements. The input is generated by repetitive peripheral magnetic stimulation (RPMS) at the innervation zone of the paretic muscles. The stimulation leads to a motion of the activated muscles. The proprioceptive input is generated by two mechanisms: adequately by activation of mechanoreceptors of the stimulated muscles during the induced contractions and relaxations and inadequately by direct activation of the involved sensorimotor afferents. The method has been applied to 52 patients suffering from spastic paresis of the upper extremity. A simple clinical quantification using the Ashworth scale revealed that spasticity could be remarkably (1-2 points) reduced already by one session of RPMS lasting 15 minutes. In order to get an objective insight into the improvement of active motor performances, a neurophysiological investigation of active finger extensions was performed in eight patients suffering from a central hemiparesis. Following RPMS of the paretic finger extensors, the patients could perform rapid finger extensions with larger displacement and velocity at diminished amounts of EMG activity. PMID- 12719623 TI - Age-related outcomes in persons with spinal cord injury: a summary paper. AB - Etiology and level of injury often discriminate between age groups for persons with spinal cord injury, complicating the understanding of what role age actually has on outcomes. The age of the patient is sometimes used as a factor in determining the appropriateness of a referral to inpatient rehabilitation. When role of age is unclear or misunderstood, though, the referral and admission decision is subject to discrimination and ageism. This paper presents information that may assist in making more appropriate decisions. By reviewing the results of four studies examining the role of age-at-injury on the outcomes of persons with spinal cord injury, including such factors of functional improvement and discharge disposition, the overall effects of age can better be understood and more appropriate conclusions drawn. PMID- 12719624 TI - Conformational changes and truncation of tau protein during tangle evolution in Alzheimer's disease. AB - The conformation-dependent antibodies Tau-66 and Alz-50 recognize discontinuous epitopes on the tau molecule (residues 155-244 & 305-314 and 5-15 & 312-322, respectively), thereby defining two distinct conformations. In double- and triple label immunofluorescence experiments we discovered that specific populations of neurofibrillary tangles display either the Alz-50 or the Tau-66 conformation, but not both. In combination with other antibodies to several domains of the molecule we demonstrate that the conformation recognized by Alz-50 seems to be an early event in the formation of neurofibrillary tangles. This conformation is characterized by the presence of predominantly intact N- and C- termini. By contrast, the Tau-66 conformation is likely a later event in tangle development, being favored in structures containing truncations of both the N- and C- termini. We propose a sequence of events that occurs during the formation and evolution of neurofibrillary tangles based on the initial conformation adopted by tau. In this scheme, the Tau-66 conformation in neurofibrillary tangles may arise from amino and carboxy truncation of tau after it has assumed the Alz-50 conformation. These results indicate that tau structure within the NFT is dynamic in that tau can undergo a "refolding" event following N- and C-terminal truncation. PMID- 12719625 TI - Catching tau in the act. PMID- 12719626 TI - Role of cytokines in the gene expression of amyloid beta-protein precursor: identification of a 5'-UTR-binding nuclear factor and its implications in Alzheimer's disease. AB - One of the major neuropathological characteristics of Alzheimer's disease (AD) is the brain depositions of senile plaques that are mainly composed of toxic amyloid beta-peptide (Abeta), which is generated from a family of Abeta containing precursor proteins (AbetaPP; 695-770 amino acids). The role of cytokines and growth factors has been implicated in the pathogenesis of AD. Our goal is to determine the mode of action of cytokines on the regulation of betaPP gene expression. Here we studied the effect of different cytokines on the activity of 5'-untranslated region (5'-UTR) of betaPP mRNA in human astrocytic cells (U-373). We compared betaPP-5'-UTR activity in the presence of interleukin-1 (IL-1alpha and IL-1beta), transforming growth factor (TGF-beta1) and tumor necrosis factor TNF-alpha1. The astrocytic cells, which were treated separately with these agents, were transfected with either the vector (pSV2CAT) or pSV2UTR-CAT construct containing 90 bp of AbetaPP 5'-UTR +54 to 144 bp). This region was cloned upstream of a reporter chloramphenicol acetyl transferase gene (CAT). Our results indicate that the treatment of pSV2UTR-CAT-transfected cells with either IL-1alpha, IL-1beta, TGF-beta1 or TNF-alpha1 stimulated reporter gene activity in a factor-specific manner. This was consistent with their effects on elevating AbetaPP protein levels. Transfection of the same cells with the pSV2CAT vector lacking 5'-UTR resulted in a reduced reporter gene activity with all treatments studied. DNA-gel shift experiments indicate that the 54/144 region binds to a nuclear protein(s) in a cell type specific manner. These results suggest that 5' UTR of the AbetaPP gene can respond to the stimulation of different cytokines, which likely regulate AbetaPP transcription and translation via regulatory elements present in the AbetaPP promoter and in 5'-UTR, respectively. The characterization of AbetaPP regulatory elements, including the 5'-UTR, will accelerate the development of novel agents against new targets for AD. PMID- 12719627 TI - Neuroprotective role of learning in dementia: a biological explanation. AB - Several epidemiological studies have found an association between low educational level (or low cognitively demanding occupations) and dementia. Although other studies have not found evidence to support such an association, there has been a general trend toward a "use it or lose it" concept which attempts to promote a neuroprotective role of intellectual activity against the development of dementia. Formation of amyloid-beta peptide (Abeta) in the brain plays a key role in the development of Alzheimer's disease whilst glutamate has been implicated in the pathophysiology of a number of neurological disorders including Alzheimer's disease and vascular dementia. Abeta can mediate neurodegeneration by a complex interaction of neurodegenerative processes that involve increasing extracellular concentration of glutamate, increasing intracellular Ca2+ concentration, and apoptosis. Long-term potentiation (LTP, a biological correlate of learning and memory) increases the sensitivity of hippocampal neurons to synaptically released glutamate whilst decreasing responses of neurons to bath applied glutamate receptor agonists and to hypoxia/ischemia in vitro. The effects of LTP are likely to involve changes in intracellular Ca2+ concentration. Based on these findings we are proposing that the LTP-induced neuroprotection in vitro may help explain the epidemiological evidence of a possible neuroprotective role of high intellectual activity against dementia. PMID- 12719628 TI - IL-10 and glucocorticoids inhibit Abeta(1-42)- and lipopolysaccharide-induced pro inflammatory cytokine and chemokine induction in the central nervous system. AB - Alzheimer's disease (AD) is characterized by neuropil threads composed of structurally abnormal neurites, neurons containing paired helical filaments of tau protein, and extracellular deposits of amyloid-beta (Abeta) peptide, a protein fragment having neurotoxic and glial immune response activating potential. In the present study, we demonstrate that an acute intracerebroventricular (icv) injection of Abeta(1-42) in the mouse induces a time- and dose-dependent production of IL-1alpha, IL-1beta, IL-6 and MCP-1 in the hippocampus and cortex as measured by ELISA. Cytokine and chemokine levels were maximal at 9 h, with MCP-1 and IL-1alpha remaining elevated over a 24 h period and IL-1beta remaining elevated over a 48 h period. The temporal profile of Abeta induced cytokine induction differed from that observed for LPS. Following an icv injection of LPS, maximal levels of IL-1alpha, IL-1beta, IL-6 and MCP-1 were attained by 9 h and, except for MCP-1, returned to levels indistinguishable from control at 24 h. MCP-1 remained elevated at 24 h and returned to basal levels at 48 h. In contrast, little production of TNF-alpha was observed under either Abeta or LPS acute stimulus conditions. Treatment with anti-inflammatory agents such as prednisolone, dexamethasone, or IL-10 inhibited both Abeta- and LPS-induced cytokine and chemokine production in the brain. In summary, icv administration of Abeta and LPS induced elevated brain levels of pro-inflammatory cytokines that could be inhibited by immune suppressing drugs and anti-inflammatory proteins, thus providing support for the utility of anti-inflammatory therapeutics in modulating the immunopathology observed in brain inflammatory diseases such as AD. PMID- 12719629 TI - Apolipoprotein E epsilon 4 in an autopsy series of various dementing disorders. AB - Apolipoprotein E (APOE) allele epsilon 4 is a risk factor for Alzheimer's disease (AD) and has also been associated with impaired recovery from brain injury. Previous studies on APOE epsilon 4 in dementing disorders other than AD have been rather conflicting, in particular concerning frontotemporal dementia (FTD) and vascular dementia (VD). In the present study we determined APOE genotype in an autopsy series of demented subjects and non-demented controls from the Netherlands Brain Bank. We attempted to create as clear-cut diagnostic groups as possible and paid close attention to AD-type histopathological changes in all cases. In comparison with the APOE epsilon 4 allele frequency in controls (0.12; n=163 subjects), the APOE epsilon 4 allele frequency was significantly increased in AD (0.42; n=320, p<0.0001), as well as in AD with Lewy bodies (0.43; n=41, p<0.0001) and in demented subjects with no other neuropathological findings than AD-histopathology insufficient for a diagnosis of AD (0.29; n=41, p<0.001). However, the APOE epsilon 4 allele frequency was not significantly increased in FTD (0.18; n=49), VD (0.10; n=20) or in Lewy body disease without concomitant AD changes (0.13; n=12). As concerns dementing disorders, our results suggest that APOE epsilon 4 is selectively associated with the presence of AD-type histopathology. PMID- 12719630 TI - Cdk5: one of the links between senile plaques and neurofibrillary tangles? AB - The relationship between amyloid plaques and neurofibrillary tangles, the two pathologic hallmarks of Alzheimer's disease (AD), is an unknown and controversial subject. However, emerging evidence from genetic and biochemical studies suggests that accumulation of amyloid beta peptides may play a causative role in AD pathogenesis. This led to the amyloid hypothesis, which proposes that amyloid beta peptides disrupt neuronal metabolic and ionic homeostasis and cause aberrant activation of kinases and/or inhibition of phosphatases. The resulting alteration in kinase and phosphatase activities ultimately leads to hyperphosphorylation of tau and formation of neurofibrillary tangles. Cyclin-dependent kinase 5 (Cdk5) is a tau kinase whose activity is induced by amyloid beta peptides. Its deregulation may represent one of the signal transduction pathways that connect amyloid beta toxicity to tau hyperphosphorylation. This article reviews the functions and regulation of Cdk5. Evidence that suggests deregulation of Cdk5 activity in AD by virtue of calpain cleavage of its activator p35 to p25 will be discussed. PMID- 12719631 TI - Alpha-synuclein aggregation and neurodegenerative diseases. AB - Alpha-synuclein is a neuronal protein originally identified in Alzheimer's disease (AD) amyloid plaques in 1993 and named non-Abeta component precursor (NACP) [92]. Later, the discovery of two missense mutations (G88C and G209A), which resulted in Ala30Pro (A30P) and Ala53Thr (A53T) substitutions, of the alpha synuclein gene in certain autosomal-dominant early onset familial Parkinson's disease (PD) has greatly promoted the understanding of the role of alpha synuclein in the pathogenesis of neurodegenerative diseases, such as PD, dementia with Lewy bodies (DLB) and multiple system atrophy (MSA) [5,6,51,75]. At present, it is widely accepted that alpha-synuclein may play a central role in several neurodegenerative disorders because of the presence of insoluble alpha-synuclein as the major fibrillar component of inclusion bodies. From the cloning of the human alpha-synuclein cDNA in 1993 to the present, alpha-synuclein has been carefully documented in many aspects. In this article, we review the progress of studies on alpha-synuclein and its role in alpha-synuclein-related neurodegenerative diseases. PMID- 12719632 TI - Von Willebrand factor promoter targets the expression of amyloid beta protein precursor to brain vascular endothelial cells of transgenic mice. AB - Dysfunction of brain vascular endothelial cells may be associated with the pathogenesis of several diseases including cerebral amyloid angiopathy, hemorrhagic stroke and Alzheimer disease. New model systems are necessary to examine the contribution of brain endothelial cells in these disorders. The Von Willebrand factor gene promoter fragment that spans sequences -487 to +247 targets the expression of LacZ marker gene in transgenic mice specifically to brain vascular endothelial cells. Transgenic mice have been prepared that express human amyloid beta protein precursor protein (AbetaPP) isoforms 695 and 751 (wild type and Dutch variant mutations) under the regulation of this VWF promoter sequence. These AbetaPP transgenes are specifically expressed in brain vascular endothelial cells. The VWF promoter is a valuable tool for targeting gene expression to brain vascular endothelial cells to provide a model to directly examine endothelial cell placement of genes and their contribution to cerebral vascular disease. PMID- 12719633 TI - Upper and lower face and ideomotor apraxia in patients with Alzheimer's disease. AB - INTRODUCTION: Apraxia of face movement in Alzheimer's disease (AD) has been rarely investigated. This study aimed at investigating the frequency of lower (mouth, tongue and throat) and upper (eyes and eyebrows) face apraxia, in AD and its relationship with limb apraxia and severity of dementia. METHODS: Fifty seven patients with AD were tested with a new standardised test of face apraxia including upper and lower face movements, which uses an item-difficulty weighted scoring procedure, the IMA test, a test of ideomotor apraxia and the M.O.D.A., a means to assess dementia severity. RESULTS: Thirteen (23%) and 19 (33%) participants were below cut-off respectively on the upper and lower face apraxia test. Both sections of the Face Apraxia Test correlated significantly with the Ideomotor Apraxia Test. However, double dissociations between different types of apraxia were observed. Both the upper and lower face apraxia tests correlated significantly with the measure of dementia severity. CONCLUSIONS: The finding show that a proportion of AD patients fails face apraxia tests. Their face apraxia is interlinked with ideomotor limb apraxia, although dissociations are possible. Severity of dementia deterioration accounts for a good proportion of the variability of AD patients' performance on face apraxia tests. PMID- 12719634 TI - Tourette syndrome and obsessive-compulsive disorder: event-related brain potentials show similar mechanisms [correction of mechansims] of frontal inhibition but dissimilar target evaluation processes. AB - OBJECTIVES: Tourette Syndrome (TS) and Obsessive-Compulsive Disorders (OCD) share many clinical similarities and show a strong comorbidity. Current theories view a frontal-striatal dysfunction as the underlying cause of many clinical aspects of both disorders. This study sought to investigate mechanisms of conceptual integration and attention in both disorders. We hypothesized that the processing of stimuli with interfering aspects would be altered in a similar way while attentional mechanisms could differ. METHODS: Event-related brain potentials (ERPs) were recorded in a modified STROOP-paradigm in groups of TS and OCD patients and in a control group. The paradigm involved the presentation of color words in a range of different colors. The subjects had to respond to words of matching word content and color and to ignore mismatching stimuli. RESULTS: Incongruent stimuli elicited a frontal negative component ("N450") which was enhanced in amplitude and prolonged in latency in both patient groups. Matching stimuli evoked enhanced N2 and P3b components representing target evaluation mechanisms. The OCD group alone displayed a larger P3b amplitude in comparison to both other groups. CONCLUSIONS: The data are interpreted to indicate that frontal inhibitory mechanisms are altered alike in TS and OCD. In contrast, only the OCD group showed evidence for aberrant target evaluation. PMID- 12719636 TI - Disturbed monitoring and response inhibition in patients with Gilles de la Tourette syndrome and co-morbid obsessive compulsive disorder. AB - OBJECTIVE: Fronto-striatal dysfunction has been discussed as underlying symptoms of Tourette syndrome (TS) with co-morbid Obsessive Compulsive Disorder (OCD). This suggests possible impairments of executive functions in this disorder, which were therefore targeted in the present study. METHODS: A comprehensive series of neuropsychological tests examining attention, memory and executive functions was performed in a group of 14 TS/OCD in co-occurrence with OCD patients and a matched control group. RESULTS: While attentional and memory mechanisms were not altered, TS/OCS patients showed deficits in executive functions predominantly in the areas of response inhibition and action monitoring. CONCLUSIONS: These findings provide further evidence for a substantial impairment of the frontal striatal-thalamic-frontal circuit. We propose that the deficits in monitoring, error detection and response inhibition constitute the major impairment of TS/OCD patients in the cognitive domain. PMID- 12719635 TI - Perceptual organization based upon spatial relationships in Alzheimer's disease. AB - Alzheimer's disease (AD) is often accompanied by impaired object recognition, thereby reducing the ability to recognize common objects and familiar faces. Impaired recognition may stem from decreased efficacy in integrating visual information. Studies of perceptual abnormalities in AD indicate an impairment in organizing elements of the visual scene, thereby confusing components of individual forms. This type of impairment is consistent with the characteristics of neural loss, which impact cortical integration. To examine the extent to which perceptual organization is impaired in AD, psychophysical measurements were made of visual perceptual grouping based upon spatial relationships in a group of AD patients and demographically matched elderly control subjects. A comparison was also made between young and elderly control subjects to evaluate the effects of aging on these capacities. Deficits in perceptual organization were found for a subgroup of AD patients, which corresponded to impairment on facial recognition. A less profound functional decline was found for the elderly control group. The degree of impairment for AD subjects did not correlate to level of dementia, but instead appears to be idiosyncratic to individual patients. These results are consistent with impaired integrative function in AD, the degree of which reflects individual differences in the regional distribution of neuropathological changes. PMID- 12719637 TI - Depression and cognitive impairment in disability-free early multiple sclerosis. AB - Cognitive and emotional capabilities were evaluated in 73 female patients with stable relapsing-remitting definite, and/or laboratory-supported multiple sclerosis (MS) and were compared with 32 matched healthy controls. Patients were categorized according to their score in the expanded disability status scale (EDSS) to either no (EDSS 0, n = 33) or few clinical signs (EDSS 1-2, n = 40) of MS without physical disability. Patients with EDSS > 0 were characterized by significantly (p < 0.001) higher scores on "von Zerssen's" depression scale, compared to controls. Patients with higher EDSS scores (1-2) showed significantly decreased performance with respect to the total score of Kimura's Recurring Figures-Test (p < 0.001), in addition. Regarding visuo-constructive functioning, patients with EDSS=0 performed to a significantly lower level (p < 0.001), compared to controls. These results indicate that depression may present as an early sign in MS followed by cognitive impairment, in particular visuo-spatial short-term memory, before physical disability appears. Neuropsychological tests as mentioned here could serve as early diagnostic tools to detect subtle disease progression and to initiate and monitor disease modifying therapies. PMID- 12719638 TI - Radical "visual capture" observed in a patient with severe visual agnosia. AB - We report the case of a 79-year-old female with visual agnosia due to brain infarction in the left posterior cerebral artery. She could recognize objects used in daily life rather well by touch (the number of objects correctly identified was 16 out of 20 presented objects), but she could not recognize them as well by vision (6 out of 20). In this case, it was expected that she would recognize them well when permitted to use touch and vision simultaneously. Our patient, however, performed poorly, producing 5 correct answers out of 20 in the Vision-and-Touch condition. It would be natural to think that visual capture functions when vision and touch provide contradictory information on concrete positions and shapes. However, in the present case, it functioned in spite of the visual deficit in recognizing objects. This should be called radical visual capture. By presenting detailed descriptions of her symptoms and neuropsychological and neuroradiological data, we clarify the characteristics of this type of capture. PMID- 12719639 TI - Opposed left and right brain hemisphere contributions to sexual drive: a multiple lesion case analysis. AB - Brain topographical studies of normal men have have shown that sexual excitation is asymmetric in the brain hemispheres. Group studies of patients with unilateral epileptic foci and other studies of patients with unilateral brain lesions have come to the same conclusion. The present study reviewed previously published single case reports of patients with frank hypo or hypersexuality subsequent to a unilateral brain lesion. Hyposexual patients tended to have left hemisphere lesions (primarily of the temporal lobe), and hypersexual patients tended to have right hemisphere lesions (primarily of the temporal lobe) (p < 0.05). We interpret this double dissociation as part of a more general phenomenon of psychic tone similarly dissociated with regard to hemispheric control, including mood, psychomotor baseline, speech rate, and even immunity. The behavioral significance of this psychic tone is to modulate approach versus avoidance behavior. PMID- 12719640 TI - Colonial allorecognition, hemolytic rejection, and viviparity in botryllid ascidians. AB - Allorecognition is a fundamental system that animals use to maintain individuality. Although embryos are usually semiallogeneic with their mother, viviparous animals are required to allow these embryos to develop inside the maternal body, but must also eliminate an "invasion" by nonself. In colonial ascidians of the family Botryllidae, when two colonies are brought into contact at their growing edges, a hemolytic rejection reaction occurs between allogeneic colonies. Morula cells, a type of hemocyte, are the major effector cells in the hemolytic rejection. Morula cells infiltrate and aggregate where the two colonies make contact, and then discharge their vacuolar contents, which contain phenoloxidase and quinones. In viviparous botryllids, colonial contact at artificially cut surfaces always results in colonial fusion and establishment of a common vascular network even between allogeneic colonies in which the growing edge contact results in rejection. This colonial fusion between incompatible colonies (surgical fusion) suggests that the allorecognition sites are not distributed in the vascular system in which the embryos are brooded. It is supposed that a common ancestor of the viviparous species lost the capacity for allorecognition in their vascular system to protect its embryos from alloreactivity, when it changed from ovoviviparous to viviparous in the course of evolution. The limited distribution of allorecognition sites would be a solution to the embryo-parent histoincompatibility in viviparity. PMID- 12719641 TI - The mind through chick eyes: memory, cognition and anticipation. AB - To understand the animal mind, we have to reconstruct how animals recognize the external world through their own eyes. For the reconstruction to be realistic, explanations must be made both in their proximate causes (brain mechanisms) as well as ultimate causes (evolutionary backgrounds). Here, we review recent advances in the behavioral, psychological, and system-neuroscience studies accomplished using the domestic chick as subjects. Diverse behavioral paradigms are compared (such as filial imprinting, sexual imprinting, one-trial passive avoidance learning, and reinforcement operant conditioning) in their behavioral characterizations (development, sensory and motor aspects of functions, fitness gains) and relevant brain mechanisms. We will stress that common brain regions are shared by these distinct paradigms, particularly those in the ventral telencephalic structures such as AIv (in the archistriatum) and LPO (in the medial striatum). Neuronal ensembles in these regions could code the chick's anticipation for forthcoming events, particularly the quality/quantity and the temporal proximity of rewards. Without the internal representation of the anticipated proximity in LPO, behavioral tolerance will be lost, and the chick makes impulsive choice for a less optimized option. Functional roles of these regions proved compatible with their anatomical counterparts in the mammalian brain, thus suggesting that the neural systems linking between the memorized past and the anticipated future have remained highly conservative through the evolution of the amniotic vertebrates during the last 300 million years. With the conservative nature in mind, research efforts should be oriented toward a unifying theory, which could explain behavioral deviations from optimized foraging, such as "naive curiosity," "contra-freeloading," "Concorde fallacy," and "altruism." PMID- 12719642 TI - Time course of protein synthesis-dependent phase of olfactory memory in the cricket Gryllus bimaculatus. AB - The cricket Gryllus bimaculatus forms a stable olfactory memory that lasts for practically a lifetime. As a first step to elucidate the cellular mechanisms of olfactory learning and memory retention in crickets, we studied the dependency of memory retention on the de novo brain protein synthesis by injecting the protein synthesis inhibitor cycloheximide (CHX) into the head capsule. Injection of CHX inhibited (3)H-leucine incorporation into brain proteins by > 90% for 3 hr. Crickets were trained to associate peppermint odor with water (reward) and vanilla odor with saline solution (non-reward) and were injected with CHX before or at different times after training. Their odor preferences were tested at 2 hr, 1 day and 4 days after training. Memory retention at 2 hr after training was unaffected by CHX injection. However, the level of retention at 1 day and 4 days after training was lowered when CHX was injected 1 hour before training or at 1 hr or 6 hr after training. To study the time course of the development of CHX sensitive memory phase, crickets that had been injected with CHX at 1 hr after training were tested at different times from 2 to 12 hr after training. The level of retention was unaffected up to 4 hr after training but significantly lowered at 5 hr after training, and the CHX-sensitive memory phase developed gradually during the next several hours. CHX dissociates two phases of olfactory memory in crickets: earlier protein synthesis-independent phase (< 4 hr) and later (> 5 hr) protein synthesis-dependent phase. PMID- 12719643 TI - Effects of lethal levels of environmental hypercapnia on cardiovascular and blood gas status in yellowtail, Seriola quinqueradiata. AB - The cardiorespiratory responses were examined in yellowtail, Seriola quinqueradiata exposed to two levels of hypercapnia (seawater equilibrated with a gas mixture containing 1% CO(2) (water PCO(2) = 7 mmHg) or 5% CO(2) (38 mmHg)) for 72 hr at 20 degrees C. Mortality was 100% within 8 hr at 5% CO(2), while no fish died at 1% CO(2). No cardiovascular variables (cardiac output, Q; heart rate, HR; stroke volume, SV and arterial blood pressure, BP) significantly changed from pre-exposure values during exposure to 1% CO(2). Arterial CO(2) partial pressure (PaCO(2)) significantly increased (P < 0.05), reaching a new steady-state level after 3 hr. Arterial blood pH (pHa) decreased initially (P < 0.05), but was subsequently restored by elevation of plasma bicarbonate ([HCO(3)( )]). Arterial O(2) partial pressure (PaO(2)), oxygen content (CaO(2)), and hematocrit (Hct) were maintained throughout the exposure period. In contrast, exposure to 5% CO(2) dramatically reduced Q (P < 0.05) through decreasing SV (P < 0.05), although HR did not change. BP was transiently elevated (P < 0.05), followed by a precipitous fall before death. The pHa was restored incompletely despite a significant increase in [HCO(3)(-)]. PaO(2) decreased only shortly before death, whereas CaO(2) kept elevated due to a large increase in Hct (P < 0.05). We tentatively conclude that cardiac failure is a primary physiological disorder that would lead to death of fish subjected to high environmental CO(2) pressures. PMID- 12719644 TI - The directional homing behaviour of the subsocial shield bug, Parastrachia japonensis (Heteroptera: Cydnidae), under different photic conditions. AB - The female subsocial shield bug, Parastrachia japonensis, provisions its nymphs by foraging on the ground in the forest during the Japanese rainy season, and the bug uses homing navigation to drag a drupe back to its burrow by the shortest route during the day. To study whether or not this bug performs this provisioning behaviour under different photic conditions, we observed the homing behaviour and homing direction of bugs in the field around the clock and/or under various weather conditions. The bugs foraged the whole day during the busiest provisioning period, and the number of walking bugs was not affected by the different weather conditions. Such navigational behaviour, regardless of the time of the day and the weather conditions, is rare in insect navigation. To test whether the bug uses visual cues, we covered the compound eyes and ocelli with opaque or clear paint just before homing began. During the day and at night, and in all weather conditions, the homing direction of blind bugs, but not those with clear-painted eyes was disoriented, indicating that this species uses visual cues dominantly under all photic conditions. PMID- 12719645 TI - Growth factor array fabrication using a color ink jet printer. AB - We have developed a novel method for growth factor analysis using a commercial color ink jet printer to fabricate substrata patterned with growth factors. We prepared substrata with insulin printed in a simple pattern or containing multiple areas of varying quantities of printed insulin. When we cultured the mouse myoblast cell line, C2C12, on the insulin-patterned substrata, the cells were grown in the same pattern with the insulin-printed pattern. Cell culture with the latter substrata demonstrated that quantity control of insulin deposition by a color ink jet printer is possible. For further applications, we developed substrata with insulin-like growth factor-I (IGF-I) and basic fibroblast growth factor (bFGF) spotted in 16 different areas in varying combinations and concentrations (growth factor array). With this growth factor array, C2C12 cells were cultured, and the onset of muscle cell differentiation was monitored for the expression of the myogenic regulator myogenin. The ratio of cells expressing myogenin varied with the doses of IGF-I and bFGF in the sections, demonstrating a feasibility of growth factor array fabrication by a color ink jet printer. Since a printer manipulates several colors, this method can be easily applied to multivariate analyses of growth factors and attachment factors affecting cell growth and differentiation. This method may provide a powerful tool for cell biology and tissue engineering, especially for stem cell research in investigating unknown conditions for differentiation. PMID- 12719646 TI - Morphological and biochemical changes in carotenoid granules in the ventral skin during growth of the Japanese newt Cynops pyrrhogaster. AB - The color of the ventral skin of the Japanese adult newt Cynops pyrrhogaster is red, whereas that of the small juvenile newts at metamorphosis is creamy. Xanthophores in the red skin have many ring carotenoid vesicles (rcv) and a few homogenous carotenoid granules (hcg), as reported earlier. To understand the reason for this change in coloration of the ventral skin of the newt, we carried out histological and biochemical studies to see whether the size and the number of carotenoid granules (hcg and rcv) in the xanthophores and also carotenoid content in the ventral skin change during the growth of this animal. By electron microscopic observation, only hcg were observed in the creamy skin of larvae at stage 59. The diameter of the hcg in the skin of the larvae was approximately 0.85 microm, but significantly decreased to 0.35 microm in the skin of the small juvenile newt. However, the number of the hcg/100 microm (2) of a xanthophore in the ventral skin was very low in the larva at stage 59, but increased in the small juvenile. The carotenoid content was very low in the creamy skin of small juveniles, but dramatically high in the red skin of the adult newts. In the red skin of the adult newt, many rcv (85%) and a few hcg (15%) were observed. However, the number of carotenoid granules (rcv and hcg)/100 microm(2) of a xanthophore in the red skin of adult newts was not different from that of hcg/100 microm (2) of a xanthophore in the creamy skin of small juveniles. The results, taken together, suggest that the increase in the size and the number of carotenoid granules and also carotenoid content in the ventral skin is very important for red body coloration during the growth of the Japanese newt Cynops pyrrhogaster. PMID- 12719647 TI - Changes in the mRNA expressions of insulin-like growth factors, their receptors, and binding proteins during the postnatal development of rat masseter muscle. AB - Morphological, biochemical, and functional changes in rat masseter muscle reportedly occur during the shift of rat feeding behavior from suckling to chewing. To determine whether insulin-like growth factors (IGFs), their receptors (IGFRs), and binding proteins (IGFBPs) are involved in the changes in rat masseter muscle during the shift of rat feeding behavior, we analyzed the expressions of IGF-I, IGF-II, IGFR1, IGFR2, and IGFBP1~6 mRNAs in rat masseter muscle between 0 and 70 days after birth using the competitive, reverse transcriptase-polymerase chain reaction (RT-PCR) method. Between 14 and 19 days of age, sharp falls in the quantities of IGF-I, IGF-II, IGFR1, IGFR2, IGFBP3, IGFBP5, and IGFBP6 mRNAs were observed, whereas the quantity of IGFBP4 mRNA rose sharply during the same period. IGFBP1 and 2 mRNAs were not detectable during the postnatal development. In the present study, the shift of rat feeding behavior from suckling to chewing occurred between 14 and 19 days of age, since the pups took residues of a pellet diet which had been dropped in a cage after 14 days of age, and we removed the pups from the dams and fed them on a pellet diet at 19 days of age. Thus, the drastic changes in the quantities of IGF, IGFR, and IGFBP mRNAs in the rat masseter muscle between 14 and 19 days of age seem to be involved in the shift of rat feeding behavior. PMID- 12719648 TI - Cloning of the genes for the pituitary glycoprotein hormone alpha and follicle stimulating hormone beta subunits in the Japanese crested ibis, Nipponia nippon. AB - We have isolated a part of the gene for the pituitary glycoprotein hormone common alpha subunit (PGHalpha) and the whole gene for the follicle-stimulating hormone beta subunit (FSHbeta) in the Japanese crested ibis (Nipponia nippon), a critically endangered bird species in East Asia. The nucleotide sequence of a part of the PGHalpha gene (5026 bp) contained three exons holding the whole coding and 3' untranslated regions, but lacked a 5' untranslated region. Its exon intron structure was similar to that in mammals, but different from that in teleosts in the location of the second intron. For the FSHbeta gene, the nucleotide sequence of 7633 bp was assembled from two phage clones. The exon intron structure of three exons and two introns was similar to that observed in mammals and teleosts. In the putative promoter region of the ibis FSHbeta gene, a progesterone responsive element (PRE)-like sequence and two AP-1 responsive element-like sequences reported in the ovine FSHbeta gene were not conserved in complete form. The increased number of ATTTA motifs in the putative 3' untranslated region in comparison with those in Japanese quail and chicken FSHbeta cDNA suggested that more rapid degradation of FSHbeta mRNA occurs in this species. Deduced amino acid sequences of the ibis PGHalpha and FSHbeta showed high similarities with those of the corresponding subunits of other avian species. This is the first report on the genomic sequences of the PGHalpha and FSHbeta in an avian species. PMID- 12719649 TI - Growth and reproduction of the sea snake, Emydocephalus ijimae, in the central Ryukyus, Japan: a mark and recapture study. AB - A mark and recapture study was carried out for three years on a population of the Ijima's sea snake, Emydocephalus ijimae, in the coastal shallow water of Zamamijima Island, central Ryukyus, Japan. The relatively high recapture (47% of 167 marked snakes) suggests that E. ijimae is a particularly philopatric, sedentary species among the sea snakes. The sex ratio (male: female), approximately 1.6:1, significantly skewed from 1:1. The growth rate in SVL declined with growth, with females thoroughly growing better than males. Males and females were estimated to begin reproductive activity in the second or third summer and the third spring after birth, respectively. Frequency of female reproduction is guessed to vary from annual to biennial, or even less frequent. PMID- 12719650 TI - The relation among emergence date, days until first effective mating, fecundity and adult longevity in Isoperla aizuana and Sweltsa sp. (Plecoptera). AB - The relation between emergence date and the factors concerned in reproduction were studied in the stoneflies Isoperla aizuana (Perlodidae) and Sweltsa sp. (Chloroperlidae) in the laboratory. Individuals emerging earlier had larger adult size and greater adult longevity. Fecundity was also related to emergence date, adult size, and adult longevity. Furthermore, individuals emerging earlier had more days until first effective mating and tended to mate more effectively than later-emerging individuals. The maturity of the reproductive tissue may differ with emergence date. While most of the later-emerging individuals had a lower rate of effective mating, longer-lived and larger size adults mated more effectively in females of Sweltsa sp. The differences in mating character might affect the effectiveness of mating. PMID- 12719651 TI - Two new marine gastrotrichs of the genus Ptychostomella (Macrodasyida, Thaumastodermatidae) from South Korea. AB - Two new thaumastodermatid gastrotrichs belonging to the genus Ptychostomella are described from the shallow sublittoral sand bottom in the eastern coast of South Korea. In having the embossed cuticular armature, Ptychostomella orientalis n. sp. is most allied to P. lepidota Clausen, 2000, the only congeneric species so far known as possessing such a cuticular armature. Ptychostomella papillata n. sp. is characterized by the terrace-shaped cuticular protrusion on head region, and the numerous papillae with sensory hair(s). Description of the two new species is prepared with a character comparison table and a key to the species of Ptychostomella. This is the first record of genus Ptychostomella from the North Pacific. PMID- 12719652 TI - Potamostoma shizunaiense gen. et sp. nov. (Nemertea: Hoplonemertea: Monostilifera): a new brackish-water nemertean from Japan. AB - Potamostoma shizunaiense gen. et sp. nov. (Nemertea: Hoplonemertea: Monostilifera) is described from the mouth of the River Shizunai, Hokkaido, Japan. This genus is readily distinguished from other monostiliferans by an oesophagus opening far anteriorly into the rhynchodaeum, a well developed excretory system extending the whole body length, terminals of the excretory collecting tubules situated between the body wall circular muscle layer and the dermis, and bilobed testes in males. PMID- 12719653 TI - Phylogenetic relationships and intraspecific variations of loaches of the genus Lefua (Balitoridae, Cypriniformes). AB - Three nominal species are known in East Asian balitorid loaches of the genus Lefua, i.e. L. echigonia, L. nikkonis, and L. costata. Lefua echigonia, with large morphological variations was recently separated into two groups, L. echigonia including the holotype and L. sp., based on morphological and ecological traits. We performed protein and DNA analyses to elucidate phylogenetic relationships among loaches of the genus Lefua and to settle the taxonomic status of L. sp. We also investigated intraspecific variations in L. echigonia s. str. to shed light on the process of formation of freshwater fish fauna in Japan. Protein analyses using two-dimensional gel electrophoresis showed that genetic distances between L. sp. and L. echigonia s. str. and between L. sp. and L. nikkonis were as large as that between L. echigonia s. str. and L. nikkonis. DNA analyses of the mitochondrial D-loop region showed that L. sp. and L. echigonia s. str. were monophyletic, respectively, while neither L. nikkonis nor L. costata was monophyletic and these species formed together a clade. The results supported the specific status of L. sp. and proposed reevaluation of the taxonomic status of L. nikkonis and L. costata. DNA analyses also showed that L. sp. was more closely related to L. echigonia s. str. than to the L. nikkonis-L. costata complex, and four local populations were distinguished in L. echigonia s. str. Distribution patterns of the four local populations of L. echigonia s. str. in Japan were approximately congruent with those of the medaka, Oryzias latipes, suggesting that differentiation in the two distantly related fishes have a common historical background. PMID- 12719654 TI - Brain-derived neurotrophic factor/TrkB signaling in memory processes. AB - Activity-dependent changes in synaptic strength are considered mechanisms underlying learning and memory. Brain-derived neurotrophic factor (BDNF) plays an important role in activity-dependent synaptic plasticity such as long-term potentiation. Recent experimental evidence supports the role of BDNF in memory processes: Memory acquisition and consolidation are associated with an increase in BDNF mRNA expression and the activation of its receptor TrkB. Genetic as well as pharmacologic deprivation of BDNF or TrkB impairs learning and memory. In a positively motivated radial arm maze test, activation of the TrkB/phosphatidylinositol-3 kinase (PI3-K) signaling pathway in the hippocampus is associated with consolidation of spatial memory through an activation of translational processes. In a negatively motivated passive avoidance test, mitogen-activated protein kinase (MAPK) is activated during acquisition of fear memory. Furthermore, recent findings suggest the importance of interaction between BDNF/TrkB signaling and NMDA receptors for spatial memory. A Src-family tyrosine kinase, Fyn plays a role in this interaction by linking TrkB with NR2B. These findings suggest that BDNF/TrkB signaling in the hippocampus plays a crucial role in learning and memory. PMID- 12719655 TI - Transient receptor potential protein as a novel non-voltage-gated Ca2+ entry channel involved in diverse pathophysiological functions. AB - In both excitable and non-excitable cells, many chemical and physical stimuli elicit continuous Ca2+ influx through yet poorly understood pathways distinct from voltage-gated Ca2+ channels, leading to activation and modulation of various cellular functions. The molecular entities of these pathways have long been enigmatic, but important clues have been obtained from recent investigations on the Drosophila transient receptor potential (TRP) protein and its mammalian homologues. TRP proteins function as non-voltage-gated Ca2+ channels that are constitutively active or gated by a multitude of stimuli including light, pheromones, lipids, temperature, acid, osmolarity, and oxidative stress; and thus they may play divergent roles in cell pathophysiology. This short paper briefly overviews the current knowledge about these channels with a main focus on their possible linkage with in vivo function. PMID- 12719656 TI - The site where newly synthesized ATP is necessary for tension development in alpha-toxin permeabilized preparations of rat proximal colon. AB - Since it was suggested in our previous study that ATP newly synthesized from ADP and phosphocreatine (PCr) by creatine kinase had an important role in Ca2+ induced phasic contraction in alpha-toxin permeabilized smooth muscle of rat proximal colon, we studied the role of newly synthesized ATP on myosin ATPase activity, by assessing a rate of force development as an index of myosin ATPase activity. The alpha-toxin-permeabilized preparations were thiophosphorylated by treatment with ATPgammaS. After the thiophosphorylation, the contraction induced by ATP plus PCr in the absence of Ca2+ reached the maximum at 30 s. When PCr was omitted from the bathing solution, the initial rate of the contraction was significantly slower, while the level of myosin light chain thiophosphorylation remained unchanged. An inhibitor of creatine kinase slowed the initial contractile rate to a rate similar to that induced by ATP alone. ADPbetaS had no effect on ATP plus PCr-induced contraction, suggesting that accumulation of ADP does not affect the initial rate of the contraction. PCr alone did not contract the thiophosphorylated-preparations. However, in the presence of ADP, PCr induced contraction at the initial rate which was slower than that induced by ATP plus PCr. These results indicate that newly synthesized ATP together with preexisting ATP is utilized as a substrate for myosin ATPase. PMID- 12719657 TI - The regulation of bone resorption in tooth formation and eruption processes in mouse alveolar crest devoid of cathepsin k. AB - Osteoclastic bone resorption has recently been implicated in the tooth formation and eruption in alveolar bone. Cathepsin K (CK) is a cysteine proteinase expressed predominantly in osteoclasts and is believed to play a critical role in degradation of bone matrix proteins. Here we present evidence that the alveolar bone resorption is essential for the tooth formation and that eruption proceeds normally in CK-deficient (CK-/-) mice. Radiographic and histological analyses revealed that the alveolar bone from these animals had no significant abnormalities during the tooth development between 5 and 28 days after birth. The tooth crown was normally erupted through the alveolar bone layer at 28 days after birth. The number of tartrate-resistant acid phosphatase-positive multinuclear cells in the alveolar bone around the tooth germ was apparently increased in 5 day-old CK-/- mice compared with age-matched littermates. More important, however, the immunohistochemical localization of matrix metalloproteinase-9 (MMP 9) was clearly increased in the CK-/- osteoclasts. In contrast, no significant difference in the immunoreactivity for cathepsin D was observed between the CK-/- osteoclasts and the wild-type ones. These results indicate that CK-/- osteoclasts are fully differentiated and are capable of degrading the organic phase of alveolar bone during the tooth formation and eruption, which may result from the compensatory action by MMP-9 increasingly expressed in the osteoclasts. PMID- 12719658 TI - Special sensitization pattern in adenosine-induced myocardial responses after thyroxine-treatment. AB - Chronic thyroxine treatment reduces the susceptibility of atrial myocardium to adenosine. While the possible role of membrane adenosine receptors in this action is supported by several studies, the involvement of intracellular adenosine mechanisms has not been defined. The present experiments were carried out in electrically driven euthyroid and hyperthyroid guinea pig atrial myocardium. The extracellular and intracellular actions of adenosine were analyzed pharmacologically by the use of specific blockers of membrane adenosine transport and intracellular adenosine deaminase (ADA). The involvement of phosphoprotein phosphatase, phospholamban, and sarcoplasmic reticulum Ca2+ ATPase (SERCA) in the adenosine-induced responses was also studied. The major findings were as follows: i) pD(2)- and E(max)-values for adenosine-induced decrease of mechanical activity were significantly reduced after an 8-day thyroxine treatment in atrial tissues; ii) in atria of thyroxine-treated animals, membrane purine transport inhibitors (dipyridamole, NBTI) induced similar leftward shifts in concentration-response curves for adenosine in both euthyroid and hyperthyroid atrial myocardium without altering the depressed E(max) values; iii) the leftward displacement evoked by inhibitors of intracellularly located ADA (coformycin, EHNA) was more striking in hyperthyroid than euthyroid myocardia. ADA inhibitors induced a complete reversal of the maximum adenosine actions; iv) inhibition by cantharidin of phosphoprotein phosphatases (after inhibition of ADA) reduced the adenosine-induced responses. This inhibition was stronger in hyperthyroid atria; v) pharmacological elimination of sarcoplasmic reticulum Ca2+ ATPase by cyclopiazonic acid did not alter the cardiac responses to adenosine and this was independent of thyroid status. It is suggested that distinct modulation of the extra- and intracellular adenosine actions is present in eu- and hyperthyroid hearts. In the latter, a predominance of intracellular adenosine mechanisms can be proposed. PMID- 12719659 TI - Effect of TTC-909 on cerebral infarction following permanent occlusion of the middle cerebral artery in stroke prone spontaneously hypertensive rats. AB - We investigated the effect of TTC-909, a drug preparation of the stable prostaglandin I(2) analogue clinprost (isocarbacyclin methylester; methyl 5 [(1S,5S,6R,7R)-7-hydroxy-6-[(E)-(S)-3-hydroxy-1-octenyl] bicyclo[3.3.0]oct-2-en-3 yl] pentanoate) incorporated into lipid microspheres, on cerebral infarction 7 days after permanent occlusion of the middle cerebral artery (MCA) in stroke prone spontaneously hypertensive rats (SHRSP). Under the anesthesia, the MCA was permanently occluded above the rhinal fissure. In schedule 1, vehicle or TTC-909 was injected i.v. once daily over 7 days starting immediately after MCA occlusion. In schedule 2, vehicle or TTC-909 was infused for 3 h starting immediately after MCA occlusion. In schedule 3, vehicle or TTC-909 was infused for 3 h starting immediately after MCA occlusion followed by bolus injection once daily over 6 days. Seven days later, the infarct volume was estimated following hematoxylin and eosin staining. Cerebral infarction produced by permanent occlusion of MCA was limited to the cerebral cortex. While this volume was reduced significantly in case of schedule 3, the infarct volume was not reduced significantly in schedules 1 and 2. Ozagrel, a thromboxane A(2) synthetase inhibitor, had no effect on the infarct volume in schedule 3. These results suggest that cerebral infarction can be developed progressively not only during the first few hours but also after a permanent occlusion of MCA in SHRSP. TTC-909 inhibited cerebral infarction, maybe by improving cerebral blood flow and by protecting against neuronal damage. PMID- 12719660 TI - Effects of chronic administration of zonisamide, an antiepileptic drug, on bone mineral density and their prevention with alfacalcidol in growing rats. AB - We investigated the effects of chronic administration of zonisamide, an antiepileptic agent, on bone metabolism in growing rats. Administration of zonisamide at a dose of 80 mg/kg per day, s.c. for 5 weeks significantly decreased bone mineral density (BMD) at the tibial metaphysis and the diaphysis. The percent rate of decrease in BMD at the tibial metaphysis and the tibial diaphysis was 9.2% and 5.0%, respectively. There was no significant difference between these groups in the growth of the rats. Treatment with zonisamide at a dose of 80 mg/kg increased serum pyridinoline level, a marker of bone resorption, while it does not affect the serum intact osteocalcin level, a marker of bone formation. Combined administration of alfacalcidol, an active vitamin D(3) metabolite, at a dose of 0.1 microg/kg per day with zonisamide prevented a decrease in BMD and showed an increase of serum pyridinoline levels. These results suggest that zonisamide may cause bone loss by accelerating bone resorption rather than inhibiting bone formation. Moreover, the bone loss induced by zonisamide could be prevented by combining zonisamide with alfacalcidol. PMID- 12719661 TI - Effects of DX-9065a, an inhibitor of factor Xa, on ellagic acid-induced plantar skin thrombosis assessed in tetrodotoxin- and N(omega)-nitro-L-arginine-treated rats. AB - Effects of DX-9065a ((+)-2S-2-[4-[[(3S)-1-acetimidoyl-3-pyrrolidinyl]oxy]phenyl] 3-[7-amidino-2-naphthyl]propanoic acid hydrochloride pentahydrate), a dibasic propanoic acid and an inhibitor of factor Xa, were compared with those of argatroban, a low molecular weight thrombin inhibitor, on the ellagic acid induced plantar skin thrombosis in the rats treated with tetrodotoxin and N(omega)-nitro-L-arginine. Plantar skin blood flow was measured with laser Doppler flow meters, and skin temperature of the hindlimb was monitored simultaneously. In order to induce thrombus in plantar skin vasculature, ellagic acid (300 microg, i.a.) was injected into a branch of femoral artery. The formation of thrombus in femoral and plantar vessels was assessed by light microscopy. Ellagic acid decreased plantar skin blood flow and skin temperature. Intravenous injections of DX-9065a (3 mg/kg) and argatroban (1-3 mg/kg) significantly inhibited the ellagic acid-induced disturbance of plantar skin blood flow and lowering skin temperature without affecting bleeding time. The oral administration of DX-9065a (30-100 mg/kg) significantly prevented the decrease in skin temperature induced by ellagic acid, but it partially inhibited the disturbance of plantar skin blood flow. DX-9065a and argatroban also prolonged prothrombin time in a dose-dependent manner. These results suggest that DX-9065a effectively prevented thrombosis produced by ellagic acid in the skin circulation without a risk of bleeding. PMID- 12719662 TI - Possible involvement of small oligomers of amyloid-beta peptides in 15-deoxy delta 12,14 prostaglandin J2-sensitive microglial activation. AB - In Alzheimer's disease, fibrillar amyloid-beta (Abeta) peptides form senile plaques associated with microglia. However, the relationship between Abeta peptides and microglia is not fully understood. In this study, the incubation of Abeta1-40 (Abeta40) produced small oligomers, while incubation with Abeta1-42 (Abeta42) caused large molecular aggregates. Microglial production of nitrite, interleukin-6 and tumor necrosis factor-alpha was induced by Abeta40, but not Abeta42. This production was significantly reduced by 15-deoxy-Delta(12,14) prostaglandin J(2), and it was completely suppressed by beta-sheet breaker peptide, Leu-Pro-Phe-Phe-Asp. These results suggest that small oligomers, rather than large molecular aggregates, mediate microglial activation induced by Abeta peptides. PMID- 12719663 TI - Changes in blood viscosity with synthetic protease inhibitors. AB - We examined the effects on whole blood viscosity and coagulation time of various dosages of the synthetic low-molecular protease inhibitors gabexate mesilate and nafamostat mesilate with an oscillation-type viscometer. When either agent was added, blood viscosity decreased dose-dependently along a sigmoid-like curve. Furthermore, coagulation time was shorter with gabexate mesilate than with nafamostat mesilate owing to the differences of half-life in human blood. Thrombin generation, which results from the activation of coagulation factors, is inhibited by synthetic protease inhibitors and subsequently decreases blood viscosity dose-dependently. PMID- 12719664 TI - [Childhood Mediterranean visceral leishmaniasis]. AB - Visceral leishmaniasis (VL) is endemic in areas bordering the Mediterranean Sea (Spain, Italy, France, Greece, Morocco, Tunisia) where it is caused by Leishmania infantum and it is transmitted by the bite of hematophagous sandfly belonging to Phlebotomus spp.; dog constitutes the main reservoir of the infection. In comparison with the past, when VL was typically observed more frequently in children, the current ratio of childhood to adult cases is approximately 1:1. The onset of the disease is characterized by a non-specific initial symptomatology; fever, pallor and splenomegaly are always present. Pancytopenia is present very often; the laboratory diagnosis is established by serological tests (indirect fluorescent-antibody assay, immunoassay test, indirect hemagglutination assay) and by demonstration of Leishmania parasites by microscopy, culture or polymerase chain reaction (PCR) in the bone marrow aspirates. The use of PCR performed on peripheral blood has been reported to be highly sensitive for the diagnosis and the follow up of children with VL. Pentavalent antimonial drugs have been used for many decades as standard treatment for VL; in Italy liposomal amphotericin B (AmBisome) is nowadays considered the first-line treatment for VL. PMID- 12719665 TI - Uselessness of liver biopsy in patients with hepatitis C virus chronic infection and persistently normal aminotransferase levels. AB - This case-control study evaluated the real need for liver biopsy in subjects with persistently normal aminotransferase values over a long period by comparing the histological features of these subjects with those of patients with abnormal aminotransferase values. We considered as "Cases" all 32 consecutive anti-HCV/HCV RNA positive subjects with at least eight normal serum ALT values during the last twelve months; for each "Case", we selected as a "Control" one anti-HCV/HCV-RNA positive patient with at least two abnormal serum ALT values during the last twelve months. The Cases and Controls were matched for age ( 5 years) and sex. In the Case group, 1 subject showed normal liver tissue, 18 minimal chronic hepatitis (CH) and 13 mild CH. In the Control group, 7 subjects showed minimal CH, 19 mild CH, 3 moderate CH, 1 severe CH and 2 cirrhosis. The subjects in the Control group showed a significantly higher HAI score (5.39+2.81) than those in the Case group (2.96+1.62, p < 0.001). The subjects in the Control group more frequently showed a fibrosis score greater than 1 (28.1%) compared to the Case group (9.4%; p<0.05). Finally, steatosis was more frequent and more severe in the Control group than in the Case group (respectively, 78.1% vs 50%, p < 0.05; and 1.47+1.16 vs 0.6+0.71, p < 0.001). The HAI and fibrosis scores did not correlate with the ALT value, HCV genotype or HCV viral load in either the Case or Control group. Our findings showed that the subjects with a persistently normal serum ALT value had minimal or mild chronic hepatitis, thus demonstrating that a liver biopsy is not indicated for these subjects. PMID- 12719666 TI - [First evaluations of LPV/RTV (Kaletra) efficacy on HIV-positive patients treated with multiple drugs]. AB - At the current epidemic stage, characterized by the rise of antiretroviral drug resistance, it is necessary to administer to HIV-positive patients increasingly effective treatments. This is possible only by means of powerful drugs. In a retrospective study, the authors evaluate 78 patients: 76 pre-treated with multiple drugs and 2 na ves. The 78 patients received LPV/RTV, starting from the fourth 3-month period of 2000 until the first 3-month period of 2002. The average treatment duration was 6.5 ( 5.5) months; the median value 6 months. The efficacy of the LPV/RTV therapeutic regimen was evaluated by a cytofluorimetric count of CD4+ and determination of the HIV viral load. There were 14 drop-out patients (17.9%): 5 because of auto-suspension, 1 due to absence of clinical and virological efficacy, 5 due to side effects (3 hepatopathy, 1 allergy and 1 nausea); three patients were lost on follow-up. There were 64 (82.1%) patients on treatment. Forty patients responded (51.3%) and 13 (16.7%) had uncontrolled viraemia (over than 200 copies/ml). However, the treatment with LPV/RTV was not interrupted for these patients, because in the follow-up they showed an increase in CD4+ values. The authors conclude that the LPV/RTV combination confirms previous findings: it is a drug with a relatively low incidence of side effects, capable of powerful results even in the treatment of patients receiving multiple drugs and thus subjected to the risk of developing antiretroviral drug resistance. PMID- 12719667 TI - [Treatment of bacterial conjuntivitis with topical ciprofloxacin and norfloxacin: a comparative study]. AB - The authors studied the epidemiology of external ocular infections and the therapeutic efficacy of topical ciprofloxacin and norfloxacin in the treatment of conjunctivitis and blepharitis. Signs and symptoms in two study-group (ciprofloxacin group A = 95 patients and norfloxacin group B = 95 patients) as well as the microbiological data obtained from the conjunctival swab before and after treatment were considered. The clinical and bacteriological success rate in the two study-groups was respectively 91% (group A) and 83% (group B). The results of the present study confirm the therapeutic efficacy of topical fluoroquinolones in the treatment of bacterial conjunctivitis and blepharitis. PMID- 12719668 TI - [HHV8 infection and acute lymphoblastic leukaemia in children]. AB - HHV8 has been consistently linked to both classical and endemic Kaposi's Sarcoma (KS), primary effusion lymphoma and multicentric Castleman's disease. HHV8 has also been associated to other oncologic diseases although such reports have not been confirmed. Little is known about the transmission routes of HHV8. The main transmission route may differ between developed and developing countries. We carried out a serologic study by Immunofluorescence of antiHHV8 antibodies on 40 children with Acute Lymphoblastic Leukaemia (ALL) and their relatives. 5 children with ALL were positive (12.5%). Seroprevalence was not significantly higher than the western Sicily pediatric population. The variation in seroprevalence between the relatives of HHV8 seropositive and seronegative patients was not significant. Therefore HHV8 does not appear to be correlated with ALL and the main transmission route in our cases could occurr outside the family. PMID- 12719670 TI - Enanthema as the first clinical manifestation of abacavir hypersensitivity reaction: a case report. AB - Abacavir is a nucleoside analogue reverse transcriptase inhibitor used in combination with other antiretroviral drugs for the treatment of HIV 1-infection. Approximately 3% of patients who receive abacavir develop an idiosyncratic hypersensitivity reaction. The most common symptoms are fever, skin rash and gastrointestinal disorders. Respiratory symptoms occurred in approximately 20% of patients who have hypersensitivity reaction. We describe the first case, to our knowledge, of hypesensitivity reaction characterized by enanthema and fever without skin rash promptly resolved after discontinuation of abacavir PMID- 12719669 TI - [Subphrenic abscess during the course of acute brucellosis: diagnosis and treatment with percutaneous sonographically-guided drainage]. AB - Abscess formation during the course of acute brucellosis is a rare event. A case of subphrenic abscess, the first to our knowledge, is described. A 49 years-old male patient with fever and a mild increase in ALT and gamma-GT was referred to our Institution. Routine blood exams tested negative and antibodies against Brucella spp. Were also negative. CT examination of abdomen was normal. After 8 days, US examination showed a liquid area under the right diaphragma and US guided puncture revealed an abscess; pus culture showed the presence of Brucella melitensis. Seven days later theWright reaction became positive. After percutaneous catheter drainage of the abscess, fever disappeared and US follow-up showed reconstitution of subphrenic space. Our study confirm that sonography is a valid method to demonstrate abdominal abscess and that US-guided percutaneous puncture and drainage are useful tools in diagnosis and treatment of fluid abdominal collections PMID- 12719671 TI - [Cutaneous cryptococcosis in an immunocompetent host]. AB - An immunocompetent 62-year-old woman presented with cutaneous lesions over her right arm and the hypogastrium. The diagnosis of cutaneous cryptococcosis was made on the basis of the demonstration and isolation of Cryptococcus neoformans from the aspirated pus. The patient responded to fluconazole treatment. PMID- 12719672 TI - Does coinfection with Human Herpesvirus 6 have a protective effect for HIV-1 progression? PMID- 12719673 TI - [Observations on the 1348 plague epidemic. Measures taken to combat its tragic effects and avoid epidemic recrudescence]. AB - When the "Black Death" swept through Europe from southern France in 1348, in the short space of two years the Europeans were hit by one of the most serious epidemics ever recorded in human history. Yersinia pestis reached Europe by sea, its contamination propagated by the Genoese ships coming from the Crimean port of Jaffa. For the first time the world experienced microbiological unification: East and West were equally involved in the tragedy that spread, and no town remained unscathed during the various epidemic waves which succeeded one another in the following three centuries. The authors of this article describe how and why the epidemic spread, as well as the factors that led to the swift, and often fatal, involment of millions of Europeans. The second part of the article deals with the measures taken by the healthcare authorities of European towns and countries in order to halt the proliferation of the disease. According to the data and observations by authoritative authors, selected among the many who studied the disease that from the 14th century spread like a scourge throughout the known world at the time, the epidemic could have been even more serious, in terms of mortality and morbidity, without the disciplinary and provisional health measures taken. The experience gained in Italy and all over Europe at the time proved useful not only to better manage the epidemics which cyclically broke out, but also to efficiently combat the cholera epidemics of the 19th century. With the 14th century plague epidemic, the Europeans and their political and administrative representatives may well have realized for the very first time that contamination could be combatted by adopting a set of rational, scientific norms - although in practice such rules were mostly inspired by misguided scientific theories. Humankind was no longer alone. A new society was emerging, one that was not going to passively accept the more or less mysterious ways of a superior being of fate. The Italian and German city-states, the emerging nations (France, England, Austria, Spain, Holland) developed and adopted procedures to control the epidemic - at the first on an ad hoc basis, then permanently. Sometimes success was achieved but always at a high price on the part of the individual and the local community. The path to an epidemic-free existence for humankind was still a long one. PMID- 12719674 TI - A guest editorial: STEM. PMID- 12719675 TI - Data monitoring and safety committees and their operations. AB - The scientific burden of proof for new therapies and technologies in obstetrics and gynecology has risen steadily over the past 15 to 20 years, and the randomized clinical trial has become integral to this higher standard. Data monitoring and safety committees are now critical components of clinical trials, yet their history, rationale, methods of operation, and overall role have not been well-characterized in the obstetrics and gynecology literature. In this review, an overview of the important and evolving role of data monitoring and safety committees is provided. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader will be able to describe what a data monitoring and safety committee is, plan who should comprise the DMSC, and outline the functions of a DMSC. PMID- 12719676 TI - Cesarean delivery for fetal distress: rate and risk factors. AB - The objective of this article was to review the recent English language literature on cesarean delivery for fetal distress to determine its incidence, diagnostic tests, and the contributing factors to this obstetric complications. A PubMed search (1990-2000) with items of "cesarean, fetal distress," "cesarean, non-reassuring fetal heart rate," "cesarean, neonatal acidosis," and "cesarean, umbilical arterial pH," was undertaken. Reports, letters to the editor, focus on anomalous fetuses, and papers not specifically focused on this topic were excluded. Of the 392 articles that the search yielded, 169 met the inclusion criteria. Based on 37 reports with more than 1,000 patients each, the overall risk of prompt cesarean delivery for fetal concern was 3.1% (43,340 of 13,989,74). The risk exceeded 20% in patients with moderate/severe asthma, severe hypothyroidism, severe preeclampsia, and postterm or fetal growth restricted fetuses with abnormal Doppler studies. Use of likelihood ratios suggests that Doppler of the umbilical artery is a superior diagnostic test to amniotic fluid index in identifying parturients at risk for cesarean for non-reassuring fetal heart rate tracing. Although several risk factors increase the need for cesarean delivery for fetal distress, in general, most are unpreventable. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader will be able to estimate the incidence of cesarean delivery for non-reassuring fetal heart rate tracing, outline potential diagnostic tests that are useful for the detection of fetal distress, and summarize medical and obstetric conditions that place patients at risk for cesarean delivery for fetal distress. PMID- 12719677 TI - Common complementary and alternative therapies for yeast vaginitis and bacterial vaginosis: a systematic review. AB - This article is a systematic review of the literature regarding the most commonly used complementary and alternative medicine (CAM) therapies for yeast vaginitis and bacterial vaginosis. A search was conducted of all published literature on conventional search engines (PubMed, EMBASE, the Cochrane Registry, CINAHL, LILACS) and alternative medicine databases (Natural Medicines Comprehensive Database, Longwood Herbal Taskforce, and Alternative Medicine Alert), for all studies of the five most commonly used CAM treatments of vaginitis. Inconsistencies in definition of vaginitis, type of intervention, control groups, and outcomes prevented performance of a meta-analysis, and paucity of high quality studies made ranking by evidence-based scales unsuitable. Lactobacillus recolonization (via yogurt or capsules) shows promise for the treatment of both yeast vaginitis and bacterial vaginosis with little potential for harm. Boric acid can be recommended to women with recurrent vulvovaginal Candidal infections who are resistant to conventional therapies, but can occasionally cause vaginal burning. Because of associated risks in the absence of well-documented clinical benefits, douching remains a practice that should not be recommended for the treatment of vaginitis. Finally, tea tree oil and garlic show some in vitro potential for the treatment of vaginitis, but the lack of in vivo studies preclude their recommendation to patients for the time-being. The available evidence for CAM treatments of vaginitis is of poor quality despite the prevalent use of these therapies. Well-designed randomized, controlled trials investigating the efficacy and safety of these therapies for vaginitis are needed before any reliable clinical recommendations can be made. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader will be able to list the most common complementary and alternative medicine therapies for vaginitis, summarize the data surrounding the efficacy of each therapy, describe the adverse affects of each therapy, and outline which therapies are recommended and not recommended for vaginitis. PMID- 12719678 TI - Delicate balance between theory and practice: health status assessment and item response theory. PMID- 12719679 TI - Quality assessment tools: incremental advance or paradigm shift? PMID- 12719680 TI - Measuring disparity among American Indians and Alaska Natives; who's counting whom? PMID- 12719681 TI - Interpretation of changes in health-related quality of life: the remarkable universality of half a standard deviation. AB - BACKGROUND: A number of studies have computed the minimally important difference (MID) for health-related quality of life instruments. OBJECTIVE: To determine whether there is consistency in the magnitude of MID estimates from different instruments. METHODS: We conducted a systematic review of the literature to identify studies that computed an MID and contained sufficient information to compute an effect size (ES). Thirty-eight studies fulfilled the criteria, resulting in 62 ESs. RESULTS: For all but 6 studies, the MID estimates were close to one half a SD (mean = 0.495, SD = 0.155). There was no consistent relationship with factors such as disease-specific or generic instrument or the number of response options. Negative changes were not associated with larger ESs. Population-based estimation procedures and brief follow-up were associated with smaller ESs, and acute conditions with larger ESs. An explanation for this consistency is that research in psychology has shown that the limit of people's ability to discriminate over a wide range of tasks is approximately 1 part in 7, which is very close to half a SD. CONCLUSION: In most circumstances, the threshold of discrimination for changes in health-related quality of life for chronic diseases appears to be approximately half a SD. PMID- 12719682 TI - Simple as possible? Or too simple? Possible limits to the universality of the one half standard deviation. PMID- 12719683 TI - Interpreting health-related quality of life scores: the simple rule of seven may not be so simple. PMID- 12719684 TI - Is it simple or simplistic? PMID- 12719685 TI - Measuring disability in Medicare home care patients: application of Rasch modeling to the outcome and assessment information set. AB - BACKGROUND: The Outcome and Assessment Information Set (OASIS) is the universal clinical assessment tool for adult nonmaternity patients receiving skilled care at home from Medicare-certified home health agencies in the United States. Anticipating increased use of OASIS data for research purposes, this article explored the usefulness of Rasch modeling to address disability measurement challenges presented by the unique response category structure of the seven activities of daily living (ADL) and eight instrumental ADL (IADL) items in the OASIS. OBJECTIVES: To illustrate how Rasch model statistics can be used to evaluate OASIS ADL and IADL item unidimensionality and model fit; to illustrate how Rasch modeling simultaneously estimates ADL and IADL item difficulty, thresholds between item response categories, and person disability; and to compare Rasch estimates of item difficulty and person disability scores to estimates based on more conventional Likert scoring techniques. SUBJECTS: Medicare-eligible home health care patients (n = 583) served by one of 12 home care agencies in Ohio between November 1999 and September 2000. MEASURES: ADL and IADL items were measured three ways: according to the original OASIS scoring (raw Likert); transformed raw Likert scores accounting for the nonuniform item structure (corrected Likert); and Rasch Partial Credit model scores. RESULTS: The items bathing and telephone use showed evidence of unexpected response patterns; recoding of these items was necessary for good Rasch model fit. Partial Credit model results revealed that interval distances between response categories varied widely across the 15 ADL and IADL items. When ADL and IADL items were ranked by level of difficulty, results were similar between Rasch and corrected Likert measurement approaches; however, corrected Likert person scores were found to be nonlinear at highest and lowest disability levels when plotted against Rasch person scores. CONCLUSIONS: Rasch modeling can help improve the precision of disability measurement in Medicare home care patients when using ADL and IADL items from the OASIS instrument. PMID- 12719686 TI - Quality of health care for women: a demonstration of the quality assessment tools system. AB - BACKGROUND: Consumers, purchasers, and regulators are seeking information on quality for a variety of purposes. To address these demands, methods are required that are flexible in meeting the information needs of different audiences. OBJECTIVES: To test a new clinically detailed, comprehensive approach to quality measurement called Quality Assessment (QA) Tools. DESIGN: Quality measures were developed for women ages 18 to 50 years for preventive care and 17 clinical areas that included chronic and acute health problems. A stratified random sample of women enrolled in 1 of 2 health plans in 1996 to 1997 was drawn and data abstracted from the medical records of all their providers for a 2-year period. FINDINGS: We evaluated quality for 758 women in 2 managed care plans. Quality of care varied substantially depending on the dimension being examined. For example, acute care was significantly better than chronic or preventive care. Quality was highest for follow-up care and lowest for treatment in both plans. Quality by modality ranged from approximately 90% for referral or admission to 16% for education and counseling. We found significant differences between the plans in the quality of care for 7 of the 17 conditions studied. CONCLUSION: The QA Tools system offers an alternative approach to evaluating health system performance. Potential advantages include the richness of the information produced by the system, the ability to create summary scores for consumers and purchasers, and the system-level performance information for use in quality improvement activities. PMID- 12719687 TI - Disparities in hospitalizations of rural American Indians. AB - BACKGROUND: Disparities in hospitalization rates, particularly rates for avoidable hospitalizations, are indicators of potentially unmet health needs and inefficient use of health resources. Hospitalization rates that the Indian Health Service (IHS) can report underestimate disparities for American Indians (AIs) and Alaska Natives (ANs) relative to other Americans, because the IHS cannot track all hospitalizations of AIs/ANs in their user population. OBJECTIVES: To compare hospitalization and avoidable hospitalization rates for a rural AI/AN user population with those of non-Indians living in the same counties where both groups use the same hospital system, regardless of the expected source of payment. RESEARCH DESIGN: Retrospective analysis of California hospital discharge data for 1996 linked to rural IHS user data for 1995 and 1996 (3920 hospitalizations) compared with a random sample of discharge data for the rest of the non-Indian population in the 37 counties of the IHS Contract Health Service delivery area (7840 hospitalizations). MEASURES: Hospitalization and avoidable hospitalization rates and risk ratios (RRs). RESULTS: Hospitalization and avoidable hospitalization rates were both higher for the AI/AN user population than for the non-Indian general population. The age-adjusted hospitalization ratios were 72% higher for men (RR 1.72, confidence interval [CI] 1.40-2.12) and 52% higher for women (RR 1.52, CI 1.36-1.92). The comparable ratios for avoidable hospitalizations were 136% higher for men (RR 2.36, CI 1.52-3.29) and 106% higher for women (RR 2.06, CI 1.32-3.50). CONCLUSIONS: Disparities in both hospitalization and avoidable hospitalization rates of rural AIs/ANs in California were previously undetected by either federal IHS or state hospital discharge data alone. At least some of the disparities are likely reducible with improved access to care. PMID- 12719688 TI - Are critical pathways effective for reducing postoperative length of stay? AB - BACKGROUND: Many hospitals use critical pathways to attempt to reduce postoperative length of stay (PLOS) for diverse conditions and procedures. OBJECTIVE: To evaluate whether critical pathways were associated with reductions in postoperative PLOS after accounting for prepathway trends in PLOS. RESEARCH DESIGN: Retrospective cohort study, from 1988 to 1998. SETTING: Academic medical center department of surgery. SUBJECTS: A total of 10,960 admissions eligible for 1 of 26 critical pathways implemented from 1990 to 1996, from 2 years before to 2 years after each pathway implementation date. Coding definitions were developed and validated to identify admissions eligible for each pathway, and data were abstracted from the hospital's discharge database. MEASURE: A pathway was considered effective if, after its implementation, there was a statistically significant decrease in the prepathway trend for PLOS. RESULTS: Median number of annual eligible admissions per pathway was 59 (range, 18-706). Median PLOS for the prepathway periods was 8 days (interquartile range, 5-10 days). For 16 (62%) pathways, PLOS was already declining in the prepathway period. After adjusting for demographics, comorbidity, admission characteristics, and prepathway time trends in PLOS, 7 (27%) pathways were associated with a significant postimplementation decrease in the rate of change in PLOS (range among the 7 pathways, 5-45% decrease) and none with a significant increase from the prepathway trend for PLOS. CONCLUSION: Critical pathways may decrease postoperative stay for some, but not all, surgeries. Trends toward decreasing length of stay over time may reduce the impact of critical pathways on this outcome. PMID- 12719689 TI - Relationship of employee-reported work limitations to work productivity. AB - BACKGROUND: Work limitation rates are crucial indicators of the health status of working people. If related to work productivity, work limitation rates may also supply important information about the economic burden of illness. OBJECTIVE: Our objective was to assess the productivity impact of on-the-job work limitations due to employees' physical or mental health problems. RESEARCH DESIGN: Subjects were asked to complete a self-administered survey on the job during 3 consecutive months. Using robust regression analysis, we tested the relationship of objectively-measured work productivity to employee-reported work limitations. SUBJECTS: We attempted to survey employees of a large firm within 3 different jobs. The survey response rate was 2245 (85.9%). Full survey and productivity data were available for 1827 respondents. MEASURES: Each survey included a validated self-report instrument, the Work Limitations Questionnaire (WLQ). The firm provided objective, employee-level work productivity data. RESULTS: In adjusted regression analyses (n = 1827), employee work productivity (measured as the log of units produced/hour) was significantly associated with 3 dimensions of work limitations: limitations handling the job's time and scheduling demands (P = 0.003), physical job demands (P = 0.001), and output demands (P = 0.006). For every 10% increase in on-the-job work limitations reported on each of the 3 WLQ scales, work productivity declined approximately 4 to 5%. CONCLUSION: Employee work limitations have a negative impact on work productivity. Employee assessments of their work limitations supply important proxies for the economic burden of health problems. PMID- 12719690 TI - Is the prevalence of gatekeeping in a community associated with individual trust in medical care? AB - BACKGROUND: Consumer concerns about the restrictions of managed care may lead to distrust. OBJECTIVES: To examine whether a community's level of gatekeeping activity is associated with an individual's trust in medical care. RESEARCH DESIGN: Cross-sectional cohort (N = 49,929). SUBJECTS: Participants in a nationally representative sample derived from the Community Tracking Survey who had health insurance, had a usual source of care, made at least 1 physician visit, and resided in one of the sampled metropolitan areas with corresponding community-level data, including the prevalence of gatekeeping activity. MEASURES: Four questions measuring trust in physician. RESULTS: Individuals from communities with a higher prevalence of gatekeeping activity report less trust than individuals from areas with a lower prevalence of gatekeeping activity, after adjusting for whether that individual had a health plan with a gatekeeper requirement. For example, in communities with the highest prevalence of gatekeeping activity relative to the lowest, the odds ratio for individuals to agree strongly that they trusted their doctor to put their medical needs above all other considerations was 0.77 (95% confidence interval, 0.71-0.84). Also, a higher prevalence of gatekeeping in the community was positively associated with the perception that a physician was strongly influenced by insurance company rules when making decisions about medical care. Conversely, a higher prevalence of gatekeeping in the community was negatively associated with the perception that a doctor might perform an unnecessary test or procedure and with concern about restricted referral for specialty care. CONCLUSION: Individuals' trust in their physicians may be influenced by wider contextual variables, like the prevalence of gatekeeping in the community. PMID- 12719691 TI - Measuring the quality of depression care in a large integrated health system. AB - BACKGROUND: Guideline-based depression process measures provide a powerful way to monitor depression care and target areas needing improvement. OBJECTIVES: To assess the adequacy of depression care in the Veterans Health Administration (VHA) using guideline-based process measures derived from administrative and centralized pharmacy records, and to identify patient and provider characteristics associated with adequate depression care. RESEARCH DESIGN: This is a cohort study of patients from 14 VHA hospitals in the Northeastern United States which relied on existing databases. Subject eligibility criteria: at least one depression diagnosis during 1999, neither schizophrenia nor bipolar disease, and at least one antidepressant prescribed in the VHA during the period of depression care profiling (June 1, 1999 through August 31, 1999). Depression care was evaluated with process measures defined from the 1997 VHA depression guidelines: antidepressant dosage and duration adequacy. We used multivariable regression to identify patient and provider characteristics predicting adequate care. SUBJECTS: There were 12,678 patients eligible for depression care profiling. RESULTS: Adequate dosage was identified in 90%; 45% of patients had adequate duration of antidepressants. Significant patient and provider characteristics predicting inadequate depression care were younger age (<65), black race, and treatment exclusively in primary care. CONCLUSIONS: Under treatment of depression exists in the VHA, despite considerable mental health access and generous pharmacy benefits. Certain patient populations may be at higher risk for inadequate depression care. More work is needed to align current practice with best-practice guidelines and to identify optimal ways of using available data sources to monitor depression care quality. PMID- 12719692 TI - Analyzing hospital length of stay: mean or median regression? AB - BACKGROUND: Length of stay (LOS) is an important measure of hospital activity and health care utilization, but its empirical distribution is often positively skewed. OBJECTIVE: This study reviews the mean and median regression approaches for analyzing LOS, which have implications for service planning, resource allocation, and bed utilization. METHODS: The two approaches are applied to analyze hospital discharge data on cesarean delivery. Both models adjust for patient and health-related characteristics, and for the dependency of LOS outcomes nested within hospitals. The estimation methods are also compared in a simulation study. RESULTS: For the empirical application, the mean regression results are somewhat sensitive to the magnitude of trimming chosen. The identified factors from median regression, namely number of diagnoses, number of procedures, and payment classification, are robust to high-LOS outliers. The simulation experiment shows that median regression can outperform mean regression even when the response variable is moderately positively skewed. CONCLUSION: Median regression appears to be a suitable alternative to analyze the clustered and positively skewed LOS, without transforming and trimming the data arbitrarily. PMID- 12719693 TI - Trends in knee magnetic resonance imaging. AB - OBJECTIVE: To assess trends in knee magnetic resonance imaging (MRI) use and accompanying management changes to determine whether indications for this test have changed over time. DATA SOURCES: Large administrative database containing health care information for 587,010 people living in 1 state who were enrolled in the Medicare or Medicaid programs. They all had used health services during 3 consecutive years between 1991 to 1995. METHODS: We performed a retrospective cohort study examining the rates of lower extremity MRI in successive years and calculated the proportion of patients who were seen by a knee specialist or underwent knee surgery subsequent to the MRI. RESULTS: The annual rate of knee MRI was 1.4 per 1000 person-years in 1991 and increased by 140% to 3.4 per 1000 person-years by 1995 (P = 0.001). Approximately half of patients who underwent a knee MRI in 1991 had a diagnosis of internal derangement of the knee in the prior year; this figure dropped to 35% in 1995 (P < 0.001). The percentage of patients undergoing a knee MRI who had no record of any knee diagnosis in the prior year grew from 13% in 1991 to 33% in 1995 (P < 0.001). Over the 5 years of the study, the percentage of patients whose knee MRI was followed by specialist care or knee surgery decreased from 68% to 58%, a relative change of 15% (P < 0.005). CONCLUSION: Knee MRI use increased sharply during the study period, but the proportion of such patients who had a prior diagnosis of internal derangement or subsequently saw a knee specialist or underwent knee surgery decreased. This finding indicates that the criteria for knee MRI appear to have broadened substantially during this period. PMID- 12719694 TI - Estimation of excess risk of readmission to hospital after an index inpatient separation. AB - OBJECTIVES: To develop methods to measure excess risk of readmission following an index admission using linked administrative health data. RESEARCH DESIGN: The cumulative risk of readmission following an index admission was calculated for index and reference subjects using linked hospital morbidity, death, and electoral roll data in cohort, cohort-crossover, and cohort-comparison-crossover designs. SUBJECTS: Index subjects were defined as any man age 20 years or older who separated from an acute hospital in Western Australia in 1990 to 1995 following any form of prostatectomy for a principal diagnosis of benign prostatic hypertrophy. Reference subjects were selected from the general population and the electoral roll (cohort designs). Cases were also used as their own historical controls (cohort-crossover) with and without adjustment for background time difference (cohort-comparison-crossover). MEASURES: The excess risk of readmission following an index admission was estimated by calculating the cumulative risk of readmission in index subjects and subtracting the background risk of admission. The background risk calculation varied according to the study design. RESULTS: The risk of readmission at 30 days increased by 241 to 328% following the procedure. After 1 year of follow-up, the risk of readmission was still increased by 58 to 108%. In general, the absolute differences between index and reference subjects decrease or remain the same with increasing rigor of the methods. CONCLUSIONS: In this example, there was little difference between the cohort-crossover and the cohort-comparison-crossover designs, suggesting that the cohort-crossover method is a justifiable method in the absence of electoral roll controls. PMID- 12719695 TI - Eletriptan (relpax) for migraine. PMID- 12719696 TI - Insulin-sensitizing drugs for polycystic ovary syndrome. PMID- 12719697 TI - Biochemical diagnosis of gastroenteropancreatic endocrine tumors. AB - Functioning gastroenteropancreatic endocrine tumors produce and secrete different substances that can be detected in the plasma and cause hormone-related syndromes. Symptoms such as diarrhea associated either with typical skin rash or peptic ulcer disease may be suggestive of the presence of intestinal carcinoid or gastrinoma. Other clinical manifestations such as severe hypoglycemia, diabetes, necrolytic erythema and gallbladder disease may also indicate an endocrine tumor. Sometimes, patients present no, or just vague, symptoms such as dyspepsia or abdominal pain and nonfunctioning endocrine tumors in these patients can be found incidentally during diagnostic imaging procedures or at operation. Usually, the diagnosis is established by the measurement of the specific tumor marker in the plasma and, sometimes, in the urine. In some cases, normal basal hormone levels are observed even in the presence of typical symptoms. Therefore, stimulatory tests such as the secretin test for gastrinomas are required to establish the diagnosis. General markers for the diagnosis of gastroenteropancreatic endocrine tumors are also available. Among these, chromogranin A has proved to be of great value for diagnosing nonfunctioning tumors and is considered the most sensitive general marker. The availability of both specific and general markers as well as stimulatory tests may enable the clinician to diagnose functioning gastroenteropancreatic endocrine tumors at an early stage and to recognize nonfunctioning tumors. PMID- 12719698 TI - Prevention of type 2 diabetes: are we ready? AB - Type 2 diabetes is the most common metabolic disease. The cost of diabetes to the individual and to society, and the pandemic prevalence makes disease prevention of extreme importance. Persons with impaired glucose tolerance, modest elevations in blood glucose that remain below levels diagnostic for diabetes, are at increased risk of progression to overt diabetes. New studies evaluate the role of lifestyle interventions including diet and exercise, and the potential role of multiple classes of pharmaceutical agents including the insulin sensitizers, biguanides and thiazolidendiones, and carbohydrase and lipase inhibitors in disease prevention of such high-risk individuals. Many of these strategies appear to be effective to delay, and perhaps prevent, the development of type 2 diabetes and thus should be considered for broader clinical application. Awareness of the extent of the problem and the potential benefits of prevention needs to be raised in both physicians and in the at high risk population. PMID- 12719699 TI - [The hemophagocytic syndrome (macrophage activation syndrome)]. AB - The hemophagocytic syndrome (HPS) is an uncommon, often misdiagnosed life threatening disorder of immune regulation, characterized by a widespread proliferation and multisystemic infiltration of non-malignant histiocytes that undergo uncontrolled hemophagocytosis in bone marrow and/or reticulo-endothelial system. The HPS immune dysfunction consists in a low or absent cytotoxic T and natural killercell activity and in hyperactivation of T lymphocytes and macrophages, with consequent proinflammatory cytokine storm. Clinically, HPS is characterized by high fever, lymphadenopathies, hepato-splenomegaly, liver dysfunction, (pan)cytopenia, hyperferritinemia, hypertriglyceridemia, hypofibrinogenemia, as well as coagulopathy and neurological manifestations in many cases. A hereditary/primary variant (familial hemophagocytic lymphohistiocytosis) and an acquired/secondary form (reactive HPS) are known. This latter may occur in several disorders, including infections, immunodeficiency states, malignancies, lymphoproliferative and autoimmune diseases. Without treatment, HPS fatally has an unfavourable prognosis. Recently, outcome improvements have been reported, due to better therapeutic strategies. The clinical and pathological features of this syndrome are reviewed. PMID- 12719700 TI - [Evaluation of the adherence to the asthma guidelines by the administration of a questionnaire in community pharmacies]. AB - BACKGROUND: Several studies demonstrate that the adherence to asthma guidelines (GL) is poor, but only a few of them were performed in community pharmacies. Thus, we decided to study this phenomenon by administering a questionnaire (Q) in two pharmacies. METHODS: A Q was developed and administered to 138 patients customers of two community pharmacies in Rome. RESULTS: The severity of the disease was established based on the frequency of daytime and nocturnal symptoms before therapy, following the stepwise approach recommended by the current GL. We observed up to 90 different treatments, while those listed by the GL, long-term preventive or quick-relief for the four categories of asthma, are only 19. In particular, many of them included antihistaminic drugs and mucoactive agents, pharmacological classes not recommended by the GL, while certain long-term preventive therapeutic schemes did not include glucocorticoids. CONCLUSIONS: The analysis of the effects on daytime and nocturnal symptoms and the interferences of the disease with school and work, showed that the control of asthma was inadequate, probably due to the low adherence to the GL. In conclusion, this small pilot study, which does have several methodological limitations (small population and geographic area, small number of pharmacies involved, the use of a Q to obtain information not easily reported by the patient) confirms the important role played by pharmacists in the analysis of the adherence of pharmacological treatments to official GL. PMID- 12719701 TI - [Falls in the elderly]. AB - Falls are the main causes of accidental death and disability in elderly people, since they may especially cause hip fractures and a number of related complications. Available incidence estimates are surely in defect, because falls are often omitted both by patients, their family and caregivers. Risk factors may be classified in intrinsic and extrinsic; the former include muscular and osteoarticular diseases or other favouring conditions, whilst the latter include environmental or iatrogenic factors, such as drugs or alcohol consumption. Extrinsic factors may be rapidly modified in the elderly and thus prevented. In fact, prevention of falls is the main intervention of geriatrist, both at patient's home and if patient is hospitalized. In order to reduce the risk of falls, it is sometimes sufficient to stop a treatment or to reduce the doses of drugs causing sedation or orthostatic hypotension, to avoid if possible, the use of sedative-hypnotics, to use non-pharmacological methods for treating insomnia. The introduction of the necessary changes in the environment, the promotion of physical activity, the individuation of the subjects with a high risk of falls and the use of hip protectors are useful means for preventing falls and avoiding their harmful consequences. PMID- 12719702 TI - Detection of chromosome 1p deletion using FISH on meningioma touch imprints suggest a region outside chromosome 22 as important in tumor recurrence. AB - BACKGROUND: Atypical and anaplastic meningiomas tend to recur and to invade adjacent brain, bone, and skin. They can also metastasize to extracranial organs such as the lung, liver, or bone causing death. The classic genetic abnormality is loss in chromosome 22. Recent reports have indicated that allelic deletion of chromosome 1p is associated with malignant progression of meningiomas. METHODS: Cytogenetic analysis of 4 meningiomas was performed using double target fluorescent in situ hybridization and focusing on chromosomes 1 and 22. The meningioma series included 4 patients whose tumors were histologically benign. RESULTS: One patient's tumor had recurred. FISH was performed on 500 nuclei/tumors. All our cases showed a loss of chromosome 22q while only 1 meningioma showed an additional loss of chromosome 1p, and this was the recurred one. CONCLUSIONS: The results of this study support the existence of tumor suppressor gene(s) on 1p associated with recurrence in meningiomas and suggest that status of chromosome 1p by FISH may assist physicians in predicting prognosis of patients affected by this tumor. However more in-vestigation is needed in this direction, as our re-sults refer to a small number of subjects studied. PMID- 12719703 TI - Involvement of the nervous system and HLA phenotype in primary Sjogren's syndrome. PMID- 12719704 TI - Pilot trial of intravenous infusion of a replication-selective adenovirus (ONYX 015) in combination with chemotherapy or IL-2 treatment in refractory cancer patients. AB - ONYX-015 is an adenovirus that selectively replicates in p53 dysfunctional or mutated malignant cells. We performed a pilot trial to determine the safety and feasibility of treatment with ONYX-015 delivered intravenously in patients with advanced malignancy. One cohort of five patients received ONYX-015 once a week for 6 weeks at a dose of 2 x 10(12) particles per infusion in combination with weekly infusions of irinotecan (CPT11, 125 mg per week) and 5-fluorouracil (5FU, 500 mg per week). A second cohort of five patients received the combination of ONYX-015 at a dose of 2 x 10(11) particles per week for 6 weeks in combination with interleukin 2 (IL 2, 1.1 x 10(6) units daily via subcutaneous injection for 5 days each week for 4 weeks). Toxicity attributable to ONYX-015 was limited to transient fever. All patients demonstrated elevations in neutralizing antibody titers within 4 weeks of the infusion of ONYX-015. Serum levels of IL-6, IL-10, tumor necrosis factor-alpha, and interferon-gamma increased within 6 hours of viral infusion, suggesting immune activation. This response was more pronounced in the cohort of patients who received 2 x 10(12) particles per infusion. Two patients demonstrated uptake of viral particles in malignant tissue by quantitative PCR. Electron microscopy confirmed selective cytoplasmic viral particles within malignant cells but not within adjacent normal tissue in a third patient. In conclusion ONYX-015 can be administered safely in combination with CPT11, 5FU or low-dose IL 2 and is able to access malignant tissue following intravenous infusion. Further investigation of ONYX-015, possibly with agents that may modulate replication activity, or duration of virus survival, is indicated. PMID- 12719705 TI - Adenovirus-mediated gene transfer of enhanced Herpes simplex virus thymidine kinase mutants improves prodrug-mediated tumor cell killing. AB - The Herpes simplex virus 1 (HSV) thymidine kinase (tk) suicide gene together with ganciclovir (GCV) have been successfully used for the in vivo treatment of various solid tumors and for the ablation of unwanted transfused stem cells in recent clinical trials. With the aim of improving this therapeutic system, we compared the potential efficacy of adenoviral (Ad) vectors expressing enhanced tk mutants in vitro and in vivo. The previously created HSV-tk mutants dm30 and sr39, created by random sequence mutagenesis, were inserted into a standard Ad.RSV E1(-)E3(-) backbone using homologous recombination. GCV killing of Ad.HSV tk, Ad.dm30-tk and Ad.sr39-tk was assessed in various tumor cell lines with a cell proliferation assay. Cells expressing the two TK mutants were two-to-five fold more sensitive to GCV when compared with Ad.HSV-tk transduced cells in all cell lines tested (five human mesotheliomas, one human lung cancer, a human cervical carcinoma, a mouse fibrosarcoma, and a rat glioma line) at equal TK expression levels. Flank tumor models, including cell-mixing studies, assessed the in vivo efficacy of the engineered viruses in BALB/C and SCID mice. In all animal studies, Ad.dm30-tk and Ad.sr39-tk showed more tumor growth inhibition than Ad.HSV-tk when GCV was administered. The use of adenovirus-mediated gene transfer of both tk mutants dm30-tk and sr39-tk for cancer suicide gene therapy should provide a more effective and safer alternative to wild-type HSV-tk. PMID- 12719706 TI - Signal sequence deletion and fusion to tetanus toxoid epitope augment antitumor immune responses to a human carcinoembryonic antigen (CEA) plasmid DNA vaccine in a murine test system. AB - Carcinoembryonic antigen (CEA, CEACAM5) is expressed on several human carcinomas including colon cancer. CEA contains signal peptides that target the protein through the endoplasmic reticulum and to the cell membrane. We constructed a plasmid DNA vaccine encoding a truncated CEA (deltaCEA), devoid of its signal peptides, and demonstrated that it was retained inside the cell, while full length CEA (wtCEA) was expressed on the membrane. We hypothesized that intracellular retention of deltaCEA would enhance MHC class I presentation of CEA peptides, thus favoring cellular immune responses. In addition, a promiscuous T helper epitope (Q830-L844 of tetanus toxoid) was fused to the N-terminal of the truncated CEA gene (tetdeltaCEA). C57BL/6 mice immunized with DNA encoding wtCEA or tetdeltaCEA developed both humoral and cellular immune responses to CEA. SCID mice transplanted with spleen cells from tetdeltaCEA but not wtCEA-immunized C57BL/6 mice showed strong suppression of tumor growth after inoculation of human CEA-expressing colon carcinoma cells. Immune spleen cell populations depleted for either B, T or both B and T cells were active, indicating that effector cells might also reside in other populations. The present approach to manipulating antigen presentation may open new possibilities for immunotherapy against colon and other CEA-secreting carcinomas. PMID- 12719707 TI - Replication of an integrin targeted conditionally replicating adenovirus on primary ovarian cancer spheroids. AB - Replication competent viruses hold promise for treatment of advanced cancers resistant to available therapeutic modalities. Although preliminary clinical results have substantiated their efficacy, preclinical development of these novel approaches is limited by assay substrates. The evaluation of candidate agents could be confounded by differences between primary tumor cells and tumor cell lines, as discordance in the levels of surface receptors relevant for viral entry has been reported. Since primary tumor cells are difficult to analyze ex vivo for longitudinal observation of virus replication, we developed three-dimensional aggregates or spheroids of unpassaged and purified ovarian cancer cells as a means for prolonging primary tumor cell viability and as a three-dimensional in vitro model for replicative viral infection. Ovarian cancer cells purified from ascites samples were sustained for 30 days while retaining the infection profile with tropism modified and unmodified adenoviruses (Ads). Cell line and primary cell spheroids were used to quantitate the replication and oncolytic potency of replicative Ads in preclinical testing for human ovarian cancer trials. Therefore, spheroids provide a method to sustain purified unpassaged primary ovarian cancer cells for extended periods and to allow evaluation of replicative viruses in a three-dimensional model. PMID- 12719708 TI - Cytotoxic effect of replication-competent adenoviral vectors carrying L-plastin promoter regulated E1A and cytosine deaminase genes in cancers of the breast, ovary and colon. AB - Prodrug activating transcription unit gene therapy is one of several promising approaches to cancer gene therapy. Combining that approach with conditionally replication-competent viral vectors that are truly tumor specific has been an important objective of recent work. In this study, we report the construction of a new conditionally replication-competent bicistronic adenoviral vector in which the cytosine deaminase (CD) gene and the E1a gene are driven by the L-plastin tumor-specific promoter (AdLpCDIRESE1a). A similar vector driven by the CMV promoter has also been constructed (AdCMVCDIRESE1a) as a control. We have carried out in vitro cytotoxicity in carcinomas of the breast, ovary and colon, and in vivo efficacy studies with these vectors in an animal model of colon cancer. While the addition of the AdLpCDIRESE1a vector to established cancer cell lines showed significant cytotoxicity in tumor cells derived from carcinomas of the breast (MCF-7), colon (HTB-38) and ovary (Ovcar 5), no significant toxicity was seen in explant cultures of normal human mammary epithelial cells (HMEC) exposed to this vector. The addition of 5-fluorocytosine (5FC) significantly increased the cytotoxicity in an additive fashion of both the AdLpCDIRESE1a and AdCMVCDIRESE1a vectors as well as that of the AdLpCD replication incompetent vector to established tumor cell lines. However, no significant cytotoxicity was observed with the addition of 5FC to explant cultures of normal human mammary epithelial cells that had been exposed to the L-plastin-driven vectors. Studies with mixtures of infected and uninfected tumor cell lines showed that the established cancer cell lines infected with the AdLpCDIRESE1a vector generated significant toxicity to surrounding uninfected cells (the "bystander effect") even at a ratio of 0.25 of infected cells to infected + uninfected cells in the presence of 5FC. The injection of the AdLpCDIRESE1a vector into subcutaneous deposits of human tumor nodules in the nude mice was potentiated by administering 5-FC by intraperitoneal injection. This treatment resulted in a decreased tumor size and a decreased tumor cell growth rate. The mice treated with a combination of the AdLpCDIRESE1a vector intratumoral injection and intraperitoneal 5FC injections lived much longer than the other experimental groups exposed to the viral vector alone or to the combination of the intratumoral AdLpCD replication incompetent vector injections plus intraperitoneal 5-FC injections. These encouraging results with our newly constructed AdLpCDIRESE1a vector suggest a need for further study of its utility in a preclinical model of intracavitary therapy of pleural or peritoneal carcinomatosis. PMID- 12719709 TI - Transplantation of prodrug-converting neural progenitor cells for brain tumor therapy. AB - Since neural progenitor cells can engraft stably into brain tumors and differentiate along the neuronal and glial line, we tested the hypothesis that transplanted cytosine deaminase (CD)-expressing ST14A cells (an immortalized neural progenitor cell line) can convert locally 5-fluorocytosine (5-FC) into 5 fluorouracil (5-FU) and produce a regression of glioma tumors. ST14A, retrovirally transduced with the E. coli CD gene, showed a strong bystander effect on glioma cells as assessed by in vitro assay. Intracerebral injection of C6 glioma cells generated a rapidly growing tumoral mass. DiI prelabeled ST14A, coinjected into the rat brain with C6 glioma cells, survived in the tumoral mass up to 10 days and their number was not affected by in vivo 5-FC treatment. In contrast, a significant decrease of the glioma tumoral mass (-50%) was observed in 5-FC-treated rats. 5-FC had no effect on the tumor in the absence of CD expressing ST14A cells. Our results support the feasibility of systems based on intratumoral transplantation of prodrug-converting cells for brain tumor therapy. PMID- 12719710 TI - Triplex-forming oligodeoxynucleotides targeting survivin inhibit proliferation and induce apoptosis of human lung carcinoma cells. AB - Survivin is expressed in most cancers but is undetectable in differentiated adult cells, and plays an important role both in the suppression of apoptosis and mitotic spindle checkpoint; thus it has attracted great interest as a potential drug target. In this study, we investigated the antigene and antiproliferative effects of triplex-forming oligodeoxynucleotides (TFO) targeting survivin in human lung carcinoma A549 cells. Survivin-specific TFOs form stable triplexes under physiological conditions as tested by electrophoretic mobility shift assays. Treatment of A549 cells with survivin-specific but not control TFOs at a concentration of 400 nM in the presence of uptake-enhancing liposome significantly reduced survivin protein level, inhibited cell proliferation, and induced cell apoptosis as demonstrated by immunoblot, cell number counting, and Annexin V-staining. Moreover, we found that the triplex-forming potential of TFOs measured in vitro does not necessarily correlate with the ability of TFOs to affect expression of a targeted gene in vivo. Our results indicate that targeting survivin is a promising alternative strategy for the development of novel anticancer therapeutics. PMID- 12719711 TI - Human U251MG glioma cells expressing the membrane form of macrophage colony stimulating factor (mM-CSF) are killed by human monocytes in vitro and are rejected within immunodeficient mice via paraptosis that is associated with increased expression of three different heat shock proteins. AB - Human U251MG glioma cells retrovirally transduced with the human gene for the membrane form of macrophage colony-stimulating factor (mM-CSF) were investigated. The clones, MG-2F11 and MG-2C4, that expressed the most mM-CSF, but not the viral vector or the parental U251MG cells, were killed by both murine and human monocyte/macrophages in cytotoxicity assays. MG-2F11 cells failed to form subcutaneous tumors in either nude or NIH-bg-nu-xidBR mice, while mice inoculated with the U251MG viral vector (MG-VV) cells developed tumors. Electron microscopy studies showed that 4 hours after subcutaneous injection, the mM-CSF-transduced cells began dying of a process that resembled paraptosis. The dying tumor cells were swollen and had extensive vacuolization of their mitochondria and endoplasm reticulum. This killing process was complete within 24 hours. Macrophage-like cells were immediately adjacent to the killed MG-2F11 cells. Immunohistological staining for the heat shock proteins HSP60, HSP70 and GRP94 (gp96) showed that 18 hours after inoculation into nude mice, the MG-2F11 injection site was two to four times more intensely stained than the MG-VV cells. This study shows that human gliomas transduced with mM-CSF have the potential to be used as a safe live tumor cell vaccine. PMID- 12719712 TI - Gene transduction efficiency and maturation status in mouse bone marrow-derived dendritic cells infected with conventional or RGD fiber-mutant adenovirus vectors. AB - Since dendritic cells (DCs) play a critical role in establishing antigen-specific adaptive immune responses, in the past several years, therapeutic strategies using genetically modified DCs against cancer and infectious diseases have attracted increasing attention. In the present study, we demonstrated that RGD fiber-mutant adenovirus vector (AdRGD) exhibited markedly superior gene transduction efficiency in mouse bone marrow-derived DCs (mBM-DCs) compared to conventional adenovirus vector (Ad). Likewise, this vector exhibited superior major histocompatibility complex class I-restricted presentation of antigen derived from the delivered gene in mBM-DCs. In order to investigate the effect of Ad-infection on the DC-differentiation process (maturation), we used three types of AdRGD and three conventional Ad to transduce mBM-DCs. These vectors carried either no transgene, LacZ gene, or gp100 gene. Infection by any of the Ad vectors enhanced the expression of MHC class II molecules in mBM-DCs. CD80, CD86, and CD40 expression and IL-12 production were more efficient in AdRGD-infected mBM DCs than in conventional Ad-infected cells. Contrary to our expectations, endocytotic activity of mBM-DCs decreased only slightly upon Ad-infection, whereas antigen uptake by lipopolysaccharide (LPS)-driven mature mBM-DCs was significantly impaired. However, our reverse transcription-polymerase chain reaction analysis revealed that Ad-infection resulted in the upregulation of the chemokine receptor CCR7 and downregulation of CCR6 in mBM-DCs and LPS-stimulated cells. We, therefore, concluded that Ad-infection directly influenced DC maturation, although the effects were milder than under LPS-stimulation. In addition, this change in the immunologic properties of DCs resulted primarily from an increase in the number of Ad-particles capable of invading the cells rather than from the expression of foreign genes. AdRGD-infection caused greater induction of maturation than conventional Ad-infection, irrespective of the type of transgene inserted. PMID- 12719713 TI - Comments on augmentation of local antitumor immunity in liver by interleukin-2 gene transfer via portal vein: a possible explanation for contradictory in vivo and vitro results of interleukin-2 treatment in a rat model of colon carcinoma metastasis. PMID- 12719714 TI - p53: 25 years of research and more questions to answer. PMID- 12719715 TI - Regulation of p53 responses by post-translational modifications. PMID- 12719716 TI - Transcriptional repression mediated by the p53 tumour suppressor. PMID- 12719717 TI - Renewing the debate over the p53 apoptotic response. PMID- 12719718 TI - p53 downstream targets and chemosensitivity. PMID- 12719719 TI - Nitric oxide: a key regulator of myeloid inflammatory cell apoptosis. AB - Apoptosis of inflammatory cells is a critical event in the resolution of inflammation, as failure to undergo this form of cell death leads to increased tissue damage and exacerbation of the inflammatory response. Many factors are able to influence the rate of apoptosis in neutrophils, eosinophils, monocytes and macrophages. Among these is the signalling molecule nitric oxide (NO), which possesses both anti- and proapoptotic properties, depending on the concentration and flux of NO, and also the source from which NO is derived. This review summarises the differential effects of NO on inflammatory cell apoptosis and outlines potential mechanisms that have been proposed to explain such actions. PMID- 12719720 TI - Decision making by p53: life, death and cancer. AB - The p53 tumor-suppressor plays a critical role in the prevention of human cancer. In the absence of cellular stress, the p53 protein is maintained at low steady state levels and exerts very little, if any, effect on cell fate. However, in response to various types of stress, p53 becomes activated; this is reflected in elevated protein levels, as well as augmented biochemical capabilities. As a consequence of p53 activation, cells can undergo marked phenotypic changes, ranging from increased DNA repair to senescence and apoptosis. This review deals with the mechanisms that underlie the apoptotic activities of p53, as well as the complex interactions between p53 and central regulatory signaling networks. In p53-mediated apoptosis, the major role is played by the ability of p53 to transactivate specific target genes. The choice of particular subsets of target genes, dictated by covalent p53 modifications and protein-protein interactions, can make the difference between life and apoptotic death of a cell. In addition, transcriptional repression of antiapoptotic genes, as well as transcription independent activities of p53, can also contribute to the apoptotic effects of p53. Regarding the crosstalk between p53 and signaling networks, this review focuses on the interplay between p53 and two pivotal regulatory proteins: beta catenin and Akt/PKB. Both proteins can regulate p53 as well as be regulated by it. In addition, p53 interacts with the GSK-3beta kinase, which serves as a link between Akt and beta-catenin. This review discusses how the functional balance between these different interactions might dictate the likelihood of a given cell to become cancerous or be eliminated from the replicative pool, resulting in suppression of cancer. PMID- 12719721 TI - Mouse models with modified p53 sequences to study cancer and ageing. AB - Experiments with p53 transgenic and p53 gene-targeted mouse strains have substantiated, and in some cases challenged, a number of hypotheses on the biology of the p53 protein. New questions have emerged regarding similarities and differences between murine and human genetic networks in various tissues. Mouse models with targeted p53 alleles are now applied not only to investigate tumour susceptibility, but also to address questions pertinent to molecular epidemiology, chemoprevention, development of anticancer p53-specific pharmaceuticals, and ageing. PMID- 12719722 TI - p53 triggers apoptosis in oncogene-expressing fibroblasts by the induction of Noxa and mitochondrial Bax translocation. AB - The mechanism of p53-dependent apoptosis is still only partly defined. Using early-passage embryonic fibroblasts (MEF) from wild-type (wt), p53(-/-) and bax( /-) mice, we observe a p53-dependent translocation of Bax to the mitochondria and a release of mitochondrial Cytochrome c during stress-induced apoptosis. These events proceed independent of zVAD-inhibitable caspase activation, are not prevented by dominant negative FADD (DN-FADD), but are negatively regulated by Mdm-2. Bcl-x(L) expression prevents the release of mitochondrial Cytochrome c and apoptosis, but not Bax translocation. At a single-cell level, enforced expression of p53 is sufficient to induce Bax translocation and Cytochrome c release. Real time RT-PCR analysis reveals a significant induction of RNA expression of Noxa and Bax in p53(+/+), but not in p53(-/-) MEF. Noxa protein expression becomes detectable prior to Bax translocation, and downregulation of endogenous Noxa by RNA interference protects wt MEF against p53-dependent apoptosis. Hence, in oncogene-expressing MEF p53 induces apoptosis by BH3 protein-dependent caspase activation. PMID- 12719723 TI - Combined p53/Bax mutation results in extremely poor prognosis in gastric carcinoma with low microsatellite instability. AB - Gastric cancer is highly refractory to DNA-damaging therapies. We therefore studied both gene mutation and protein expression of p53 and Bax in a cohort of 116 patients with gastric cancer who underwent R0-resection with a curative intent. Bax mutation was independent from severe microsatellite instability (MSI), that is, global mismatch repair deficiency as determined by analysis of BAT-25/BAT-26 microsatellite markers. Thus, Bax-frameshift mutation is a feature of tumors with low MSI. In contrast and as expected, no p53 mutations were observed in the microsatellite instable tumors. p53 Mutation or p53 overexpression did not have an impact on disease prognosis. p53-Inactivation was, however, associated with an extremely poor prognosis in the subgroup of patients with Bax-mutated tumors. Thus, we show for the first time that the combined mutation of p53 and Bax, two key regulators of the mitochondrial apoptosis pathway, results in an extremely aggressive tumor biology and poor clinical prognosis. PMID- 12719724 TI - Nitric oxide promotes p53 nuclear retention and sensitizes neuroblastoma cells to apoptosis by ionizing radiation. AB - Nitric oxide (NO) is a potent activator of the p53 tumor suppressor protein. However, the mechanisms underlying p53 activation by NO have not been fully elucidated. We previously reported that a rapid downregulation of Mdm2 by NO may contribute to the early phase of p53 activation. Here we show that NO promotes p53 nuclear retention and inhibits Mdm2-mediated p53 nuclear export. NO induces phosphorylation of p53 on serine 15, which does not require ATM but rather appears to depend on the ATM-related ATR kinase. An ATR-kinase dead mutant or caffeine, which blocks the kinase activity of ATR, effectively abolishes the ability of NO to cause p53 nuclear retention, concomitant with its inhibition of p53 serine 15 phosphorylation. Of note, NO enhances markedly the ability of low dose ionizing radiation to elicit apoptotic killing of neuroblastoma cells expressing cytoplasmic wild-type p53. These findings imply that, through augmenting p53 nuclear retention, NO can sensitize tumor cells to p53-dependent apoptosis. Thus, NO donors may potentially increase the efficacy of radiotherapy for treatment of certain types of cancer. PMID- 12719726 TI - Circle of life. PMID- 12719725 TI - Mutation of p53 and consecutive selective drug resistance in B-CLL occurs as a consequence of prior DNA-damaging chemotherapy. AB - Inactivation of p53 has been shown to correlate with poor prognosis and drug resistance in malignant tumors. Nevertheless, few reports have directly shown such effects in primary tumor cells. Here, we investigated the p53 mutational status in 138 B-CLL samples and compared these findings with drug and gamma irradiation sensitivity profiles. p53 mutations resulted not only in a shorter survival but, notably also in selective resistance to alkylating agents, fludarabine and gamma-irradiation. In contrast, no such effect was observed for vincristine, anthracyclines and glucocorticoids. Thus, these latter compounds induce cell death at least in part by p53-independent pathways. Interestingly, p53 mutations clustered in patients who had received prior chemotherapy. In fact, we show for the first time that treatment with DNA-damaging alkylating agents correlates with occurrence of p53 mutations in a clinical setting. This finding may explain at least to some extent the development of resistance to second-line anticancer chemotherapy. PMID- 12719728 TI - Delivering the kiss of death. AB - Cytotoxic T lymphocytes and natural killer cells kill their targets by secreting specialized granules that contain potent cytotoxic molecules. Through the study of rare immunodeficiency diseases in which this granule pathway of killing is impaired, proteins such as Rab27a have been identified as components of the secretory machinery of these killer cells. Recent evidence suggests that the destruction of activated lymphocytes through granule-mediated killing may be an important mechanism of immunological homeostasis. Although the process by which this occurs is not yet known, it is possible that events taking place at the immunological synapse may render the killer cell susceptible to fratricidal attack by other killer cells. PMID- 12719729 TI - The death effector domain protein family: regulators of cellular homeostasis. AB - The death effector domain (DED) occurs in proteins that regulate programmed cell death. Both pro- and anti-apoptotic proteins containing DEDs have been identified. For Fas and possibly other death receptors, homotypic DED interactions connect the Fas-associated death domain (FADD) protein to caspase-8 and caspase-10 to mediate formation of the death-inducing signal complex. This complex can be inhibited by other DED-containing proteins. Accumulating evidence now suggests that DED-containing proteins have additional roles in controlling pathways of cellular activation and proliferation. Thus, the DED defines a family of proteins that may be pivotal to cellular homeostasis by establishing a 'cell renewal set point' that coregulates proliferation and apoptosis in parallel. PMID- 12719730 TI - Apoptosis in the development and maintenance of the immune system. AB - Programmed cell death is essential for the development and maintenance of cellular homeostasis of the immune system. The Bcl-2 family of proteins comprises both pro-apoptotic and anti-apoptotic members. A subset of pro-apoptotic members, called 'BH3-only' proteins, share sequence homology only in the minimal death domain, designated the Bcl-2 homology 3 (BH3) domain. BH3-only proteins operate as upstream sentinels, selectively sensing both intrinsic and extrinsic death stimuli. They communicate this information to the pro-apoptotic 'multidomain' members Bax or Bak--a process that is antagonized by anti-apoptotic members of the Bcl-2 family. The functional balance of pro-apoptotic versus anti-apoptotic influences, which operates at organelles, determines whether a lymphocyte will live or die. BH3-only molecules, often working in concert, compete for downstream multidomain pro- and anti-apoptotic BCL-2 members to control serial stages of lymphocyte development and homeostasis. PMID- 12719731 TI - Caspase-independent cell death in T lymphocytes. AB - T lymphocyte death is essential for proper function of the immune system. During the decline of an immune response, most of the activated T cells die. Cell death is also responsible for eliminating autoreactive lymphocytes. Although recent studies have focused on caspase-dependent apoptotic signals, much evidence now shows that caspase- independent, necrotic cell death pathways are as important. An understanding of the molecular control of these alternative pathways is beginning to emerge. Damage of organelles including mitochondria, endoplasmic reticulum or lysozymes, leading to an increase in calcium and reactive oxygen species and the release of effector proteins, is frequently involved in caspase independent cell death. PMID- 12719732 TI - Cellular versus humoral immunology: a century-long dispute. PMID- 12719733 TI - RNA AIDS DNA. PMID- 12719734 TI - CD8 and B cell memory: same strategy, same signals. PMID- 12719735 TI - Linking Toll-like receptors to IFN-alpha/beta expression. PMID- 12719738 TI - Entitlement. PMID- 12719739 TI - An ethical assessment framework for addressing global genetic issues in clinical practice. AB - PURPOSE/OBJECTIVES: To describe the perceptions of nurses regarding the importance of each action skill listed in the Ethical Assessment Framework (EAF) to their ethical decision-making process and how prepared they were to undertake each action when confronted by moral dilemmas in clinical practice, and to identify general genetic ethical issues of concern and frequency encountered. DESIGN: Descriptive, exploratory. SAMPLE AND SETTINGS: Members of the Oncology Nursing Society's Cancer Genetics Special Interest Group (n = 34) and the International Society of Nurses in Genetics (n = 101). METHODS: Participants completed the Ethical Assessment Skills Survey and Genetic Ethical Issues Survey. MAIN RESEARCH VARIABLES: Perceptions of level of importance and preparation for each action skill in the EAF and level of concern and frequency encountered regarding ethical issues in clinical practice. FINDINGS: Each ethical action skill listed in the EAF was rated as important to the ethical decision-making process, although minimal skill level was reported in 60% of the steps. Nurses reported major concerns about the frequently encountered issues of confidentiality, managed care, and informed consent. CONCLUSIONS: The EAF proposes action skills that can assist nurses in developing expertise in ethical decision making and offers a model for addressing genetic ethical issues in clinical practice. Protection of patient confidentiality was the number one ethical concern of nurses surveyed. IMPLICATIONS FOR NURSING: Nurses are challenged to have comprehensive and current genetic knowledge, which is necessary to advocate for, educate, counsel, and support patients and families confronting difficult genetic healthcare decisions. Nurses will be able to effectively translate genetic information to patients by developing and using ethical decision-making and counseling skills. Effective measures to protect confidentiality of patient data are important to ensure that genetic information is safeguarded. PMID- 12719741 TI - Positive attitude in cancer: patients' perspectives. AB - PURPOSE/OBJECTIVES: To describe what being positive means for patients undergoing treatment for cancer. RESEARCH APPROACH: Qualitative, descriptive approach. SETTING: Specialist cancer clinic in a large metropolitan hospital in Sydney, Australia. PARTICIPANTS: 11 patients with cancer currently being treated at a cancer clinic for a variety of cancers. METHODOLOGIC APPROACH: Semistructured interviews that were audiotaped, transcribed, and thematically analyzed for content related to being positive. MAIN RESEARCH VARIABLES: Patients' definitions of positive and negative attitude, their perceptions of the importance of attitude during their cancer journey, and any factors that influenced their perceived attitude. FINDINGS: For patients, positive attitude was defined as optimism for the day and getting though everyday events of the journey by taking control rather than focusing on the future. Factors that affected patients' positive attitude were their relationships with their specialists, people around them being positive and supportive, and having a pleasant environment at home and at the treatment center. Patients found expectations of them to be positive as being detrimental. CONCLUSIONS: Patients with cancer must be positive for the present rather than the future. INTERPRETATION: Nurses need to inspire and support patients' positivity while undergoing treatment for cancer. Nurses should not force their own value system on them nor treat them differently if they do not conform to societal expectations to be positive and optimistic for the future. PMID- 12719740 TI - Continuing education: Comprehensive menopausal assessment: an approach to managing vasomotor and urogenital symptoms in breast cancer survivors. AB - PURPOSE/OBJECTIVES: To describe the development and implementation of a comprehensive menopausal assessment (CMA) and intervention program for women with a history of breast cancer. DATA SOURCES: Published articles selected from computerized databases, conference proceedings, bibliographies of pertinent articles and books, and lay publications. DATA SYNTHESIS: The CMA program consisted of a structured, comprehensive assessment of three symptoms (hot flashes, vaginal dryness, and stress urinary incontinence) and an individualized plan of education, counseling, nonestrogen treatments, psychosocial support, referrals, and follow-up. CONCLUSIONS: A structured approach to evaluating and managing vasomotor and urogenital symptoms with, for example, the CMA, may help breast cancer survivors with severe symptoms more effectively manage these symptoms than "usual care." IMPLICATIONS FOR NURSING: Nurses providing care for women with a history of breast cancer can incorporate the key elements of the CMA program into their practice to facilitate more effective management of three common menopausal symptoms that often are undertreated in this patient population. PMID- 12719742 TI - The experience of lower limb lymphedema for women after treatment for gynecologic cancer. AB - PURPOSE/OBJECTIVES: To describe women's experiences with lower limb lymphedema to inform both preventive and management clinical practices. DESIGN: A retrospective survey. SETTING: The gynecology/oncology unit of a tertiary referral women's hospital in Australia. SAMPLE: 82 women who developed lower limb lymphedema after surgical and radiation treatment for gynecologic cancers. METHODS: Structured interviews. MAIN RESEARCH VARIABLES: Psychosocial and emotional impact, physical effects, knowledge, support, treatment modalities. FINDINGS: Women identified changes in appearance and sensation in the legs and the triggers that both preceded and exacerbated symptoms. Women described seeking help and receiving inappropriate advice with as many as three assessments prior to referral to lymphedema specialists. Many women implemented self-management strategies. Lower limb lymphedema had an impact on appearance, mobility, finances, and self-image. CONCLUSIONS: Increasing longevity after gynecologic oncology treatment requires all practitioners to be aware of known or potential triggers of lower limb lymphedema and the appropriate referral and management strategies available. Women at risk need to know early signs and symptoms and where to seek early care. IMPLICATIONS FOR NURSING: The role of nursing in acute and community care of women at risk for developing lower limb lymphedema includes (a) engaging women in protecting their legs from infection or trauma pre- and postoperatively, (b) providing nursing care and education during the pre- and postoperative phases, and (c) ensuring that women being discharged are aware of early signs and symptoms of lower limb lymphedema and how to access qualified, specialized therapists so that early and effective management can be initiated. PMID- 12719743 TI - Music versus distraction for procedural pain and anxiety in patients with cancer. AB - PURPOSE/OBJECTIVES: To test the hypotheses that the effects of a music intervention are greater than those of simple distraction and that either intervention is better at controlling procedural pain and anxiety than treatment as usual. DESIGN: Randomized, controlled experiment. SETTING: A midwestern comprehensive cancer center. SAMPLE: 60 people with cancer having noxious medical procedures such as tissue biopsy or port placement or removal; 58 provided usable data. METHODS: Participants completed measures of pain and anxiety before and after their medical procedures and provided a rating of perceived control over pain and anxiety after the procedure. MAIN RESEARCH VARIABLES: Procedural pain, state anxiety, and perceived control over pain and anxiety. FINDINGS: Contrary to hypotheses, outcomes achieved with music did not differ from those achieved with simple distraction. Moreover, outcomes achieved under treatment as usual were not significantly different from those obtained with music or distraction interventions. Some patients found that the interventions were bothersome and reported that they wanted to attend to the activities of the surgeon and the medical procedure itself. CONCLUSIONS: The effects of music, distraction, and treatment as usual are equivocal. In addition, patients have individual preferences for use of distraction during painful or anxiety-provoking procedures. IMPLICATIONS FOR NURSING: Patients having noxious medical procedures should be asked about their desire to be distracted before and during the procedure and offered a strategy that is consistent with their preferences. PMID- 12719744 TI - Research and commentary: Change in exercise tolerance, activity and sleep patterns, and quality of life in patients with cancer participating in a structured exercise program. AB - PURPOSE/OBJECTIVES: To investigate the feasibility of an exercise program patterned after a phase II cardiac rehabilitation program to improve selected physiologic and psychological parameters of health in patients with cancer. DESIGN: Prospective, repeated measures study. SETTING: Two major military medical centers in the southwestern United States. SAMPLE: 62 patients diagnosed with cancer within the previous two years. Ages ranged from 24-83 (meanX = 59). Half of the participants were male and half were female. Minorities made up 29% of the sample. Participants had a wide range of cancer diagnoses and all stages of cancer. Fifteen subjects were undergoing treatment when they enrolled in the study. More than half of the subjects exercised prior to their cancer diagnoses, but fewer than half were able to resume an exercise routine following their cancer diagnoses. METHODS: Subjects met two days each week for 12 weeks for exercise and education. MAIN RESEARCH VARIABLES: Exercise tolerance as measured with a graded exercise test, activity and sleep patterns as measured with a wrist actigraph, and quality of life (QOL) as measured with the Cancer Rehabilitation Evaluation System-Short Form. FINDINGS: Significant improvements were observed over time in exercise tolerance, selected activity and sleep patterns, and QOL among the 46 (74%) subjects who completed the program. CONCLUSIONS: Patients with various types and stages of cancer can safely exercise using a cardiac rehabilitation model and can realize significant improvements in exercise tolerance, selected activity and sleep patterns, and QOL. IMPLICATIONS FOR NURSING: Most people are aware that regular exercise is part of a healthy lifestyle. After cancer diagnosis and treatment, patients experience uncertainty regarding how to resume exercise or how to begin an exercise program as part of their rehabilitation. Participation in a structured exercise program can provide patients with a safe environment within which to exercise at an intensity appropriate to their individual needs. PMID- 12719745 TI - Seventh National Conference on Cancer Nursing Research keynote address: challenges and opportunities in cancer survivorship research. PMID- 12719746 TI - Cognitive dysfunction following adjuvant chemotherapy for breast cancer: two case studies. AB - PURPOSE/OBJECTIVES: To describe the cognitive dysfunction experienced by two women after they received adjuvant chemotherapy for breast cancer and to discuss the potential role of changes in reproductive status and depression in the development of cognitive dysfunction. DATA SOURCES: Journal articles, research data, and clinical experience. DATA SYNTHESIS: Following chemotherapy, 17%-50% of women with breast cancer experience cognitive dysfunction that may include decrements in memory, attention, and psychomotor efficiency. One mechanism that may contribute to cognitive dysfunction involves changes in reproductive status resulting from chemotherapy. Additionally, the presence of depression may confound the experience of cognitive dysfunction. CONCLUSIONS: A comprehensive description of cognitive dysfunction and improved understanding of the interrelationships among cognitive dysfunction, reproductive hormone levels, and depression in women with breast cancer receiving adjuvant chemotherapy may hasten the development of interventions for the management of cognitive dysfunction. IMPLICATIONS FOR NURSING: Nurses should teach women with breast cancer and their families about the potential for cognitive dysfunction after chemotherapy so the problem can be recognized and interventions can be implemented to help women compensate for the dysfunction. PMID- 12719747 TI - Expectations and beliefs about children's cancer symptoms: perspectives of children with cancer and their families. AB - PURPOSE/OBJECTIVES: To describe the childhood cancer symptom course experienced by children with cancer from the perspectives of the children and their families. DESIGN: Longitudinal, qualitative research approach. SETTING: The participants' homes and inpatient and outpatient pediatric cancer units in western Canada. SAMPLE: A theoretical purposive sample of 39 children (4.5-18 years of age) with a variety of cancer diagnoses and their family members. METHODS: Open-ended formal interviews with children and their family members (N = 230) and participant observation of children and their family members for more than 960 hours during various periods during their illness, at various locations, and at different points in time during the study period. MAIN RESEARCH VARIABLES: Children's and their families' perspectives of cancer symptoms experienced by children with cancer. FINDINGS: Children and families had definite beliefs and expectations about the cancer symptom experiences, including (a) short-term pain for long-term gain, (b) you never get used to them, (c) they all suck, (d) it sort of helps, and (e) they are all the same but they are all different. Underpinning all of the participants' beliefs and expectations was the experience of suffering. Their beliefs and expectations contributed to and were a direct result of cancer symptoms that were ignored, unrelieved, or uncontrolled. CONCLUSIONS: Children with cancer live with symptoms on a daily basis and have experiences of unrelieved cancer symptoms. Although cancer symptoms resulted in suffering by the children and families, they accepted the symptoms as an integral part of overcoming cancer and never expected complete symptom relief. IMPLICATIONS FOR NURSING: Oncology nurses need to be more vigilant in their assessment and management of children's cancer symptoms. Further research is warranted detailing not only children's and family's beliefs and expectations of cancer symptom experiences but also nurses' understanding and interpretations of children's cancer symptom experiences. Intervention studies designed to relieve all types of cancer symptoms experienced by children must be undertaken. PMID- 12719748 TI - Role and gender differences in cancer-related distress: a comparison of survivor and caregiver self-reports. AB - PURPOSE/OBJECTIVES: To examine role and gender differences on measures of psychological distress as a consequence of dealing with cancer. DESIGN: Cross sectional, descriptive, quantitative analyses of retrospective survey data. SETTING: A large, nonprofit, volunteer-based cancer organization. The sample was recruited through an online cancer survivor's network (61%), a rural event (24%), and hospital registries (15%). SAMPLE: Convenience sample of 135 cancer survivors matched to their family caregivers (N = 270). METHODS: Dyads (i.e., survivors and family caregivers) completed matched questionnaires requesting demographic and medical information and measures of cancer-related distress. MAIN RESEARCH VARIABLES: Role (i.e., survivor or caregiver), gender, and psychological distress. FINDINGS: Caregiver means on overall psychological distress were significantly higher than those shown for survivors. Caregiver scores were significantly higher on distress for diagnosis and fear of cancer recurrence. Females scored higher than male caregivers on cancer-related anxiety, future uncertainties, fear of recurrence, and future diagnostic tests. Gender differences were not found for survivor distress. CONCLUSIONS: Results suggest a need for gender-specific, dyad-tailored cancer support services. IMPLICATIONS FOR NURSING: As expert caregivers, nurses can provide valuable assistance with the caregiving process that may decrease distress during the family's cancer experience and adaptation period. PMID- 12719749 TI - Continuing education: The emergence of thalidomide in treating advanced renal cell carcinoma. AB - PURPOSE/OBJECTIVES: To review standard and investigational treatments in advanced renal cell carcinoma, with a focus on thalidomide. DATA SOURCES: Published articles, conference proceedings, treatment guidelines, and textbooks. DATA SYNTHESIS: The prognosis for advanced renal cell carcinoma when treated with standard regimens is poor; therefore, new treatments are needed. CONCLUSIONS: Treatment with thalidomide, alone and in combination with other therapies, may improve survival for patients with advanced renal cell carcinoma. IMPLICATIONS FOR NURSING: Proactive management of adverse effects associated with thalidomide, alone and in combination, may increase patient tolerance and compliance. PMID- 12719751 TI - Oncology and nononcology nurses' spiritual well-being and attitudes toward spiritual care: a literature review. AB - PURPOSE/OBJECTIVES: To review literature about oncology and nononcology nurses' attitudes toward spiritual care and the way that their spiritual well-being influences those attitudes. DATA SOURCES: Published research and literature review articles, books, master's theses, and doctoral dissertations. DATA SYNTHESIS: Spiritual considerations play an important part in the lives of patients with cancer. Therefore, nurses must not only examine their attitudes toward spiritual care but also identify variables that influence those attitudes. One of the major variables that may influence their attitudes is their own spiritual well-being. CONCLUSIONS: A significant association exists between attitudes toward spiritual care and spiritual well-being in nurses who care for nononcology populations. However, little research examines oncology nurses' attitudes toward spiritual care and the way that their spiritual well-being affects those attitudes. IMPLICATIONS FOR NURSING: Nurse researchers need to examine the spiritual dimension of oncology nurses and the way that this influences attitudes toward spiritual caregiving. PMID- 12719750 TI - Adherence, sleep, and fatigue outcomes after adjuvant breast cancer chemotherapy: results of a feasibility intervention study. AB - PURPOSE/OBJECTIVES: To evaluate outcomes of an intervention designed to promote sleep and modify fatigue after adjuvant breast cancer chemotherapy. DESIGN: Prospective, repeated measures, quasi-experimental, feasibility study. SETTING: Midwestern urban oncology clinics. SAMPLE: 21 female participants, ages 43-66 years (meanX = 55.3) with stage I or II breast cancer status post four cycles of doxorubicin chemotherapy. Eight had four additional cycles of paclitaxel, 10 also had radiation, and 18 took tamoxifen. METHODS: each woman continued to revise her Individualized Sleep Promotion Plan (ISPP), developed during her first cycle of chemotherapy, that included sleep hygiene, relaxation therapy, stimulus control, and sleep restriction components. The daily diary, Pittsburgh Sleep Quality Index, wrist actigraph, and Piper Fatigue Scale were used for seven days 30, 60, and 90 days after the last chemotherapy treatment and one year after the first chemotherapy treatment. MAIN RESEARCH VARIABLES: Adherence and sleep and wake, fatigue, and ISPP components. FINDINGS: Adherence to the ISPP components remained high at all times (77%-88%) except for stimulus control (36%-56%). Sleep outcome means and the actigraph revealed that (a) sleep latency remained less than 30 minutes per night, (b) the time awake after sleep onset exceeded the desired less than 30 minutes per night, (c) sleep efficiency scores ranged from 82%-92%, (d) total rest time ranged from seven to eight hours per night, (e) feelings on arising ranged from 3.7-3.8 (on a 0-5 scale), (f) nighttime awakenings ranged from 10-11 per night, and (g) daytime naps ranged from 10-15 minutes in length. Fatigue remained low, from 2.9-3.5 on a 0-10 scale. CONCLUSIONS: Adherence rates remained high for most components. Sleep and wake patterns were within normal limits except for the number and duration of night awakenings. Fatigue remained low. IMPLICATIONS FOR NURSING: Future testing using an experimental design will focus on increasing ISPP adherence and decreasing nighttime awakenings. Adopting behavioral techniques to promote sleep may result in improved sleep and lower fatigue after chemotherapy. PMID- 12719753 TI - Continuing education: Oncology emergency modules: syndrome of inappropriate antidiuretic hormone. PMID- 12719752 TI - Quality of life after among ovarian germ cell cancer survivors: a narrative analysis. AB - PURPOSE/OBJECTIVES: To describe and interpret the meaning of experiences that are important to the quality of living of ovarian germ cell cancer survivors. RESEARCH APPROACH: Qualitative description within a constructivist paradigm. SETTING: 32 member sites of the Gynecologic Oncology Group and the University of Texas M.D. Anderson Cancer Center in Houston. PARTICIPANTS: 109 women between the ages of 19-64 (median age = 36) who were enrolled on prospective clinical trials of cisplatin-based chemotherapy after surgery and disease-free for at least two years. METHODOLOGIC APPROACH: As part of a larger study, narrative responses to four semistructured questions were collected at the end of a telephone interview. Using naturalistic inquiry and qualitative description techniques, content labels were assigned to units of text that seemed to encapsulate one complete thought or idea. The labeled groups were collapsed into interpreted subthemes. Finally, four general themes were constructed as representations of shared narrative responses and meanings. FINDINGS: The four constructed themes are celebrating illness, experiencing empathetic affirmation, mourning losses, and valuing illness. INTERPRETATION: In addition to measuring physical, psychological, and sexual functioning in women surviving ovarian germ cell cancer, nurses also must understand how these issues fit into their everyday lived experiences. The four themes may help clinicians and researchers to understand issues that are important to the quality of living of ovarian germ cell cancer survivors. PMID- 12719754 TI - Axial rotation of the lower human spine by rhythmic torques automatically generated at the resonant frequency. AB - Subjects sat on a 'Balans' chair supported by the shaft of a large torque generator. The lower but not the upper part of the body was free to make to/fro movements maintained by feeding back a modified velocity signal to the generator. The stiffness could be increased by the use of a position signal. Using the equations for a torsion pendulum the good linearity observed in relationship to the applied forces allowed stiffness, inertia, damping, critical damping and the damping factor to be determined in absolute terms. The method enables parameters relevant to back function and problems to be evaluated. Eleven adults acted as subjects. PMID- 12719755 TI - Angiotensin AT1 receptor antagonist losartan and the defence reaction in the anaesthetised rat. Effect on the carotid chemoreflex. AB - Modulation at the level of the nucleus tractus solitarii (NTS) appears to be an effective way of controlling cardiovascular reflexes. Angiotensin II acting on angiotensin AT1 receptors at the central nervous system appears to have an important role in these modulatory processes. The hypothalamic defence area (HDA) is a potential source of descending fibres containing angiotensin II that innervate the NTS. We investigated the effect of AT1 receptor blockade in the NTS on the response to stimulation of HDA in anaesthetised rats treated with the neuromuscular blocking agent pancuronium bromide. The characteristic increase in heart rate, blood pressure and phrenic nerve activity evoked by electrical stimulation of HDA is decreased by the microinjection of the AT1 receptor antagonist losartan into the NTS and the cardiovascular response to carotid body chemical stimulation is also reduced. These results support the hypothesis that AT1 receptors in the NTS play a role in the modulation of cardiovascular reflexes, and modify the influence exerted on the processing of these reflexes by other areas of the central nervous system. PMID- 12719756 TI - Differential chronotropic and dromotropic responses to focal stimulation of cardiac vagal ganglia in the rat. AB - Vagal cardioinhibition is exerted through a reduction not only in the heart rate but also in the rate of propagation of the cardiac action potential and in myocardial contractility. In several species, such effects can be produced independently by selective activation of ganglia in identified 'fat pads'. In this study we investigate differential control of heart rate and atrioventricular conduction by two ganglionic clusters in the rat, a species increasingly important in studies of cardiovascular control. Epicardial sites producing low threshold changes in P-P and P-R interval of the ECG in an arterially perfused preparation were explored with concentric bipolar stimulating electrodes. Stimulation sites centred on two principal ganglia, the sinoatrial (SA) ganglion at the junction of the right superior vena cava and right atrium, and the atrioventricular (AV) ganglion at the junction of the inferior pulmonary veins and left atrium. Stimulation of the SA ganglion decreased heart rate in all preparations, with little or no effect on AV conduction in one-third. Stimulation of the AV ganglion consistently slowed conduction without eliciting a comparable bradycardia. Responses survived blockade of ganglionic transmission by trimetaphan, with an enhanced chronotropic selectivity to SA ganglion stimulation, suggesting that co-excitation of preganglionic elements en passant may have contributed to the earlier mixed responses. Effective stimulation sites were precisely circumscribed and corresponded to principal ganglionic clusters confirmed histologically. We conclude that cardiac vagal ganglia in the rat show a topographical functional organisation and are amenable to investigation using the arterially perfused preparation. PMID- 12719757 TI - Vagal control of the cranial venae cavae of the rat heart. AB - The cranial venae cavae of the rat heart are composed of cardiac muscle. We tested whether the vagus nerve has an inotropic action on these blood vessels. Stimulation of right or left vagal fibres (n = 7 animals) produced a negative chronotropic and inotropic effect. Before stimulation the basal cardiac interval was 319 +/- 25 ms and the vena caval diastolic force was 1.82 +/- 0.29 mN and the systolic force was 3.28 +/- 0.39 mN. Ten second stimulation increased the cardiac interval to a maximum of 484 +/- 77 ms and reduced the systolic force significantly to 2.83 +/- 0.39 mN (two-tailed paired t test). The diastolic or baseline force was unaffected by vagal stimulation (1.85 +/- 0.29 mN). The vagal negative inotropic action took significantly longer to reach peak effect (9.5 +/- 1.0 s versus 3.2 +/- 0.9 s) and lasted longer than the chronotropic effect (20.4 +/- 2.1 s versus 10.25 +/- 1.2 s). The negative inotropic action was still observed in paced preparations and preparations with transient constant-rate tachyarrhythmias. Both the chronotropic and inotropic effects were abolished by atropine (10(-6) M) and mimicked by acetylcholine chloride (10 nM). In order to minimize an atrial contribution to the force production a more reduced preparation was used and ganglion clusters at the cavo-atrial junction were stimulated electrically (n = 4 animals). Similar negative inotropic and chronotropic effects sensitive to hexamethonium were seen. After hexamethonium administration, positive inotopic and chronotropic effects were uncovered and these were abolished by atenolol (0.1 mg %). Methylene Blue staining of the preparation at the end of the experiment showed the presence of ganglion cells at the sites of stimulation. Ganglion clusters were never seen on the venae cavae per se. The results of this investigation show that the vagus has a powerful action on the venae cavae resembling that on the atria and mediated by acetylcholine. PMID- 12719758 TI - Role of GABA and NO in the paraventricular nucleus-mediated reflex inhibition of renal sympathetic nerve activity following stimulation of right atrial receptors in the rat. AB - The aim of this study was to determine the site within the brain at which inhibition of renal sympathetic nerve activity (RSNA) occurs following right atrial receptor stimulation. The atrial receptors were stimulated by inflating a balloon at the right vena cava-atrium junction and the reflex effect was observed before and during application of neurotransmitter agonists and antagonists into the paraventricular nucleus (PVN), or intrathecally to the spinal cord. Balloon inflation reduced RSNA by 29.1 +/- 3 % without changing blood pressure in anaesthetised Wistar rats. Microinjection of the GABA(A) receptor antagonist bicuculline (0.025 mM, 100 nl) into the PVN increased RSNA by 42.3 +/- 5 % and this was changed little by balloon inflation when PVN increased RSNA by 50.6 +/- 6.3 %. Microinjection of the nitric oxide synthase (NOS) inhibitors L-NAME (0.1 mM, 100 nl) or L-NMMA (0.2 mM, 100 nl) into PVN elicited increases in RSNA of 36 +/- 8 % or 54 +/- 10 %, respectively. Balloon inflation during PVN stimulation plus NOS inhibition resulted in RSNA activity of 8 +/- 4 % or -1 +/- 1 %, respectively, compared to baseline control. Baseline RSNA was similar throughout this series of tests ranging from 9.1 +/- 1.3 to 11.5 +/- 1.1 spike counts s(-1). To rule out the possibility that the atrial reflex inhibition was in part dependent on a dopamine-mediated PVN-spinal projection pathway inhibiting RSNA at a spinal locus, a dopamine D1 receptor antagonist SCH 23390 was intrathecally applied to the spinal cord. The effect of subsequent balloon inflation on RSNA was not significantly reduced. It was concluded that atrial receptor activation causes an inhibition of RSNA at the PVN and that this effect is mediated by GABA. PMID- 12719760 TI - Activation of the renin-angiotensin system contributes to the peripheral vasoconstriction reflexly caused by stomach distension in anaesthetized pigs. AB - Gastric distension in anaesthetized pigs reflexly elicits peripheral vasoconstriction and an increase in plasma renin activity (PRA), with vagal afferent and sympathetic efferent limbs. The aim of the present study was to quantify the contribution of the renin-angiotensin system to the peripheral vasoconstriction. In pigs anaesthetized with alpha-chloralose, changes in anterior descending coronary, superior mesenteric and left external iliac blood flow caused by stomach distension before and after blockade of angiotensin II receptors with losartan were assessed using electromagnetic flowmeters. Gastric distension for periods of 30 min was performed by injecting 0.8 l warm Ringer solution into balloons positioned within the viscus. Changes in heart rate and renal blood flow were prevented by atrial pacing and injection of phentolamine into the renal arteries, and changes in regional perfusion pressure and in baroreceptor activity were minimized by aortic constriction and denervation of the carotid sinuses. PRA was assessed by radioimmunoassay of angiotensin I. Before blockade of angiotensin II receptors by administration of losartan, stomach distension decreased coronary blood flow by 14.2 % in six pigs and mesenteric and iliac blood flow by 11 % and 17.3 %, respectively, in another six pigs. After administration of losartan, these decreases were significantly reduced to 7.4 %, 6.8 % and 8.7 %, respectively. The above responses were abolished by bilateral section of the subdiaphragmatic vagal nerves. These results show that the peripheral vasoconstriction reflexly caused by stomach distension was significantly contributed to by the concomitant activation of the renin-angiotensin system. PMID- 12719759 TI - Agonist- and nerve-induced phasic activity in the isolated whole bladder of the guinea pig: evidence for two types of bladder activity. AB - Spontaneous localised propagating waves of contraction and localised stretches have been reported to occur in the isolated whole bladder of the guinea pig. The physiological role and the cellular processes underlying these events are unknown. In order to gain insight into the mechanisms generating this complex activity, experiments were performed to examine and compare the responses of the whole bladder preparation to (i) the muscarinic agonists carbachol and arecaidine, (ii) the nicotinic ligand lobeline and (iii) nerve stimulation. High concentrations of the muscarinic agonists (>3 micro M) induced a slow rise in intra-vesical pressure upon which were superimposed pressure transients, while low concentrations (< 300 nM) induced only phasic rises in pressure. One interpretation of these data is that there are two separate mechanisms activated by muscarinic agonists: one generating contracture and the other phasic activity. Immunocytochemical staining revealed M(3) muscarinic receptors on smooth muscle cells within trabeculae and a second population of positive cells in the sub urothelial layer. This observation raises the possibility that the actions of muscarinic agonists are a consequence of activating different cell types. Lobeline (1-60 micro M) activated phasic contractions but did not cause a rise in basal pressure. Atropine did not inhibit the lobeline-induced responses but abolished the muscarinic responses. Also, hexamethonium or tetrodotoxin did not affect the lobeline-induced responses. These observations suggest that the mechanism generating phasic activity is activated by a nicotinic stimulus that does not involve ganglia, nerves or the neuromuscular junction. Stimulation of the bladder nerve at frequencies between 20 and 30 Hz for 5 s resulted in a rapid rise in intra-vesical pressure. Prolonged nerve stimulation (10-200 s) at frequencies between 1 and 10 Hz activated phasic rises in pressure. Low frequency nerve stimulation increased the frequency of agonist-induced phasic activity. Thus, nerve stimulation can also produce two forms of activity and low frequency stimulation can augment the processes generating phasic activity. These observations suggest that there are two distinct types of bladder activity: global contractions involving most of the bladder wall and phasic contractions comprising propagating waves of contraction. The mechanisms generating these contractile events appear to be different and they may involve cells located in different regions of the bladder. The nature of these mechanisms and their possible physiological significance is discussed. PMID- 12719761 TI - [Ca2+]i oscillations induced by high [K+]o in acetylcholine-stimulated rat submandibular acinar cells: regulation by depolarization, cAMP and pertussis toxin. AB - Maintaining the extracellular K(+) concentration ([K(+)](o)) between 15 and 60 mM induced oscillations in the intracellular Ca(2+) concentration ([Ca(2+)](i)) in rat submandibular acinar cells during stimulation with acetylcholine (ACh, 1 micro M). These [Ca(2+)](i) oscillations were also induced by 1 micro M thapsigargin and were inhibited by 50 micro M La(3+), 1 micro M Gd(3+), or the removal of extracellular Ca(2+), indicating that the [Ca(2+)](i) oscillations were generated by store-operated Ca(2+) entry (SOC). The frequency of the ACh evoked [Ca(2+)](i) oscillations increased from 0.8 to 2.3 mHz as [K(+)](o) was increased from 15 to 50 mM. TEA (an inhibitor of K(+) channels) also induced [Ca(2+)](i) oscillations at [K(+)](o) of 4.5 or 7.5 mM in ACh-stimulated cells. These data suggest that depolarization causes [Ca(2+)](i) to oscillate in ACh stimulated submandibular acinar cells. Pertussis toxin (PTX, an inhibitor of G proteins) caused [Ca(2+)](i) to be sustained at a high level in ACh-stimulated cells at 25 mM or 60 mM [K(+)](o). This suggests that the [Ca(2+)](i) oscillations are generated by a periodic inactivation of the SOC channels via PTX sensitive G proteins, which are stimulated by depolarization. Moreover, in the presence of DBcAMP or forskolin which accumulated cAMP the frequency of the [Ca(2+)](i) oscillations remained constant (approximately 1.2 mHz) when [K(+)](o) was maintained in the range 25-60 mM. Based on these observations in ACh stimulated submandibular acinar cells, we conclude that depolarization stimulates the PTX-sensitive G proteins, which inactivate the SOC channels periodically ([Ca(2+)](i) oscillation), while hyperpolarization or PTX inhibits the G proteins, maintaining the activation of the SOC channels. Accumulation of cAMP is likely to modulate the PTX-sensitive G proteins. PMID- 12719762 TI - Nitric oxide-dependent in vitro secretion of amylase from innervated or chronically denervated parotid glands of the rat in response to isoprenaline and vasoactive intestinal peptide. AB - The basal in vitro release of amylase was similar from rat parotid lobules of innervated and chronically denervated glands and was unaffected by the inhibitors used in this study. The secretion of amylase induced by isoprenaline or vasoactive intestinal peptide (VIP) was reduced by one-third to one-half from the lobules of the innervated glands and even more so from the lobules of the denervated glands by ODQ, an inhibitor of soluble guanyl cyclase which is activated by nitric oxide (NO) and catalyses the cGMP production. The use of N (omega)-propyl-L-arginine (N-PLA) revealed that the evoked secretion of amylase in the denervated glands depended on the activity of neuronal type NO synthase to synthesize NO. Since the denervated gland is virtually devoid of NO synthase containing nerve fibres, the neuronal type NO synthase was most probably of a non neuronal source. NO-dependent amylase secretion was agonist related, since amylase secretion evoked by bethanechol and neuropeptide Y was not reduced by ODQ or N-PLA. Hence, under physiological conditions, activation of beta-adrenoceptors (sympathetic activity) and VIP receptors (parasympathetic activity) is likely to cause secretion of parotid amylase partly through a NO/cGMP-dependent intracellular pathway involving the activity of neuronal type NO synthase, possibly of acinar origin. PMID- 12719763 TI - Uterine artery function in pregnant rats fed a diet supplemented with animal lard. AB - We hypothesised that maternal uterine artery vascular dysfunction could contribute to cardiovascular dysfunction in offspring of rats fed a diet rich in fat. Sprague-Dawley rats were fed for 10 days prior to pregnancy and throughout gestation either: (a) a control breeding diet, or (b) the same diet supplemented with 20 % w/w lard, vitamins, essential micronutrients and protein to control values. At 20 days gestation vascular function was assessed in uterine arteries and third-order mesenteric arteries. Vascular reactivity in response to application of potassium, noradrenaline, the thromboxane analogue U46619, acetylcholine and nitric oxide was assessed. Maternal plasma concentrations of factors likely to contribute to endothelial dysfunction were measured. Maximum acetylcholine-induced relaxation was impaired in the mesenteric arteries of the lard-fed dams (max % relaxation: lard-fed, 69.7 +/- 6.48; control, 85.37 +/- 2.69, P = 0.03). Uterine artery vascular function was similar in the two groups (max % acetylcholine-induced relaxation: lard-fed, 73.7 +/- 4.01; control, 77.5 +/- 4.72, P = 0.98). Concentrations of plasma lipids, 8-epi-PGF(2alpha) and leptin were normal, whereas insulin and corticosterone concentrations were raised in the lard-fed group (insulin (ng ml(-1)): lard-fed, 8.04 +/- 0.47; control, 1.35 +/- 0.37, P < 0.0001; corticosterone (ng ml(-1)): lard-fed, 1164.0 +/- 170.9; control, 541.9 +/- 96.3, P = 0.005). Fetal and placental weights were reduced in lard-fed dams (fetus (g): lard-fed, 4.27 +/- 0.38; control, 2.96 +/- 0.40, P = 0.025; placenta (g): lard-fed, 0.72 +/- 0.06; control, 0.57 +/- 0.04, P = 0.05). Cardiovascular dysfunction in offspring is not associated with reduced uterine artery endothelial function but is associated with activation of the hypothalamic-pituitary-adrenal axis, hyperinsulinaemia and fetoplacental growth retardation. PMID- 12719764 TI - Age-related analysis of glucose metabolism in spontaneously hypertensive and normotensive rats. AB - This study was designed to investigate the effect of both hypertension and ageing on the efficiency of glucose metabolism. A 12-sample, 120 min intravenous glucose tolerance test (IVGTT) was applied to 36 rats: two groups of nine young (12 weeks) spontaneously hypertensive and Wistar Kyoto rats (Y-SHR and Y-WKY group, respectively) and two groups of nine old (40 weeks) SHR and WKY rats (O-SHR and O WKY group, respectively). Insulinaemia and glycaemia data were interpreted in terms of estimates of glucose effectiveness, S(G), and insulin sensitivity, S(I), provided by the minimal model of glucose kinetics. The possible link between insulin resistance and hypertension was investigated by comparing Y-SHR vs. Y-WKY and O-SHR vs. O-WKY groups. Comparison of O-SHR vs. Y-SHR and O-WKY vs. Y-WKY groups enabled us to investigate the role of age in the development of abnormalities in glucose metabolism. No significant differences (P > 0.05) were observed in the mean S(G) and S(I) estimates between SHR and age-matched WKY groups. This finding indicates that exposure of SHR to high blood pressure levels does not necessarily lead to the development of insulin resistance and impaired glucose effectiveness. Similarly, no significant differences (P > 0.05) were observed in S(G) and S(I) estimates between old and young SHR and WKY groups. This finding indicates that, in this animal model of hypertension, insulin sensitivity and glucose effectiveness do not even deteriorate with ageing. PMID- 12719765 TI - Chronic hypoxia delays myocardial lactate dehydrogenase maturation in young rats. AB - The effect of exposure to hypobaric hypoxia for 4 weeks (oxygen pressure = 106 hPa), equivalent to 5500 m in altitude) on myocardial total lactate dehydrogenase (tLDH) activity and isoform (H and M) composition was comparatively studied in growing (4.5 weeks old) and in adult (4.5 months old) male rats. The consequences of the hypoxia-induced anorexia were checked in growing rats using a pair-fed group. Exposure to hypoxia induced a significant decrease in the H/tLDH ratio in the left (LV) and right ventricle (RV) of growing and adult rats. In adult rats this alteration was mainly a consequence of the significant increase in the specific activity of the M isomer, which resulted in an increase in the overall LDH activity. In contrast, in the LV of young rats exposed to hypoxia, the specific activity of the M isomer was similar to that of normoxic animals while the H isomer activity was significantly lower than in normoxic rats, and the overall LDH activity remained unchanged. These effects were specifically due to hypoxia per se since no significant alterations were observed in pair-fed animals. In the hypertrophied RV, the alteration of H and M isomers following hypoxia was similar to that observed in adults (i.e. no change in H and an increase in M isoform). We conclude that the well-known hypoxia-induced decrease in the H/tLDH ratio is governed by different age-dependent mechanisms. In adult rats, hypoxia may induce in both ventricles a stimulating effect on M isomer expression. In the LV of growing rats this stress could inhibit the H isomer maturation without any effect on the M isomer. In the RV of growing rats this effect could have been counteracted by the growth effect of the hypertrophying process. PMID- 12719766 TI - Trimetazidine reduces basal cytosolic Ca2+ concentration during hypoxia in single Xenopus skeletal myocytes. AB - We tested the hypotheses that: (1) Ca(2+) handling and force production would be irreversibly altered in skeletal muscle during steady-state contractions when subjected to severe, prolonged hypoxia and subsequent reoxygenation; and (2) application of the cardio-protective drug trimetazidine would attenuate these alterations. Single, living skeletal muscle fibres from Xenopus laevis were injected with the Ca(2+) indicator fura 2, and incubated for 1 h prior to stimulation in 100 micro M TMZ-Ringer solution (TMZ; n = 6) or standard Ringer solution (CON; n = 6). Force and relative free cytosolic Ca(2+) concentration ([Ca(2+)](c)) were measured during continuous tetanic contractions produced every 5 s as fibres were sequentially perfused in the following manner: 3 min high extracellular P(O(2)) (159 mmHg), 15 min hypoxic perfusion (3-5 mmHg) then 3 min high P(O(2)). Hypoxia caused a decrease in force and peak [Ca(2+)](c) in both the TMZ and CON fibres, with no significant (P < 0.05) difference between groups. However, basal [Ca(2+)](c) was significantly lower during hypoxia in the TMZ group vs. the CON group. While reoxygenation generated only modest recovery of relative force and peak [Ca(2+)](c) in both groups, basal [Ca(2+)](c) remained significantly less in the TMZ group. These results demonstrated that in contracting, single skeletal muscle fibres, TMZ prevented increases in basal [Ca(2+)](c) generated during a severe hypoxic insult and subsequent reoxygenation, yet failed to protect the cell from the deleterious effects of prolonged hypoxia followed by reoxygenation. PMID- 12719767 TI - Metabolic heat production, heat loss and the circadian rhythm of body temperature in the rat. AB - Metabolic heat production (calculated from oxygen consumption), dry heat loss (measured in a calorimeter) and body temperature (measured by telemetry) were recorded simultaneously at 6 min intervals over five consecutive days in rats maintained in constant darkness. Robust circadian rhythmicity (confirmed by chi square periodogram analysis) was observed in all three variables. The rhythm of heat production was phase-advanced by about half an hour in relation to the body temperature rhythm, whereas the rhythm of heat loss was phase-delayed by about half an hour. The balance of heat production and heat loss exhibited a daily oscillation 180 deg out of phase with the oscillation in body temperature. Computations indicated that the amount of heat associated with the generation of the body temperature rhythm (1.6 kJ) corresponds to less than 1 % of the total daily energy budget (172 kJ) in this species. Because of the small magnitude of the fraction of heat balance associated with the body temperature rhythm, it is likely that the daily oscillation in heat balance has a very slow effect on body temperature, thus accounting for the 180 deg phase difference between the rhythms of heat balance and body temperature. PMID- 12719768 TI - Whole blood viscosity, plasma viscosity and erythrocyte aggregation in nine mammalian species: reference values and comparison of data. AB - In this study species-specific values for whole blood viscosity (WBV), plasma viscosity (PV) and erythrocyte aggregation (EA) were determined in a total of 360 animals. We used 40 individual adult animals of nine mammalian species: horse, pig, dog, cat, rat, cattle, sheep, rabbit and mouse. WBV measurements were carried out using a LS30 viscometer, PV was measured using OCR-D and EA was measured using a Myrenne aggregometer and the LS30 (aggregation index at low shear rate). At low shear rates (0.7 s(-1) and 2.4 s(-1)) haematocrit (Hct) standardized (40 % Hct) samples showed a higher value of WBV and EA in horse, pig, dog and cat. In cattle, sheep, rabbit and mouse, EA and WBV were markedly decreased and EA was almost undetectable, although the plasma fibrinogen concentration was higher in these animals. Rats showed the highest WBV at low shear rate in native blood and WBV was not different from horse in Hct standardized blood; however, EA was very low in the rat, a result that might be explained by mechanical or geometrical properties of the red blood cell. EA correlated with the plasma protein concentration in each species except dog and mouse. In horse, cattle and pig, EA correlated with the plasma fibrinogen concentration. At high shear rate (94 s(-1)), WBV was higher in cattle than cat and rat, and dog had higher values than horse, suggesting specific interspecies differences depending on low shear and high shear values of WBV, as a result of mechanisms that influence RBC flexibility. PV was highest in cattle and lowest in rabbit and mouse and did not correlate with WBV. Haemorheological parameters differed between the species. Each species has its own rheological fingerprint. The physiological significance of these variations among mammalian species has not yet been established. Viscosity contributes to endothelial cell shear stress. While haemorheological parameters differ across the species it may be postulated that factors influencing flow-mediated endothelial cell signal transduction are different among the species. PMID- 12719769 TI - Heart rate variability and endogenous sex hormones during the menstrual cycle in young women. AB - To our knowledge, the relationship between all four endogenous female sex hormones and resting cardiac autonomic function has not been studied. The aim of the current study was to examine the association between the normal endogenous levels of oestrogen (17beta-oestradiol), progesterone, luteinising hormone and follicle-stimulating hormone and heart rate variability (HRV) during the menstrual cycle in young eumenorrheic women. Ten healthy, young, female subjects volunteered for this study. HRV and endogenous hormone levels were recorded at three phases of the menstrual cycle: menses (day 3.8 +/- 0.5), ovulation (day 15.8 +/- 0.7) and luteal (day 22.1 +/- 0.4) to ensure HRV recordings at times of low (menses) and high (ovulation and luteal) hormonal influence. Heart rate recordings were obtained from supine resting subjects and analysed on a Holter analysis system. Total power (TP, 0-1.0 Hz), low frequency (LF, 0.041-0.15 Hz), high frequency (HF, 0.15-0.80 Hz) and LF/HF components of HRV were examined. Despite a significantly greater HR at ovulation and normal cyclic variations in all endogenous sex hormone levels, no measure of HRV was significantly different between menstrual cycle phases. Significant correlations between oestrogen levels and absolute measures of HRV at ovulation were identified. The results of the current study demonstrated that the normal cyclic variations in endogenous sex hormone levels during the menstrual cycle were not significantly associated with changes in cardiac autonomic control as measured by HRV. Significant correlation between peak oestrogen levels and HRV measures at ovulation provided further support for the reported cardioprotective effects of oestrogen in healthy females. PMID- 12719770 TI - Cardiomyocyte death and the ageing and failing heart. AB - Mammalian cardiomyocytes have limited regenerative capacity, such that cell death can result in a net loss of viable contractile elements and a decrease in cardiac functional reserve, both during normal ageing and after insults to the myocardium leading to heart failure. At least four types of cell death have been described, with apoptosis and necrosis being the most extreme phenotypes and most extensively studied. Many of the classical morphological and biochemical features associated with these forms of cell death have been derived from studies conducted in vitro and these may not always faithfully reflect events occurring in vivo. Before therapeutic interventions can be realistically developed, more studies need to be undertaken in vivo to simultaneously investigate these different death pathways, their control mechanisms and their relative contributions in depleting the pool of viable cardiomyocytes. We recently demonstrated immunohistochemically that a single injection of either a natural or synthetic catecholamine induces both cardiomyocyte apoptosis (identified by an anti-caspase 3 antibody) and necrosis (identified by an anti-myosin antibody) in the rat heart in vivo. After optimising the experimental conditions for hormone dose and temporal and spatial peaks of damage, the incidence of necrosis was 4-10 times greater than the incidence of apoptosis. Myocytes in the soleus muscle were also severely (7-10 %) damaged, involving both apoptosis and necrosis. In both striated muscles high levels of myocyte co-localisation for apoptosis and necrosis were observed, suggesting that secondary necrosis had occurred in most of the apoptotic myocytes in vivo. The ability of the catecholamines to cause myocyte death suggests that they might play an aetiological role in the progression of heart failure where over-activation of the sympathetic system results in sustained pathophysiological levels of these catecholamines. PMID- 12719771 TI - The blissful state of endothelium. PMID- 12719772 TI - Magnetic resonance direct thrombus imaging: a novel technique for imaging venous thromboemboli. AB - Invasive testing is now seldom required in patients with suspected venous thromboembolism (VTE). However, a corollary of noninvasive imaging is increased complexity as results are often yielded as probabilities rather than definitive answers and additional testing is frequently required following initial imaging. This creates a milieu in which misunderstandings and protocol violations are common, potentially leading to diagnostic errors. A highly accurate noninvasive imaging technique which allows immediate treatment decisions to be made is needed. Magnetic resonance direct thrombus imaging (MRDTI) is a novel technique which detects methaemoglobin in clot, allowing visualisation of thrombus without using intravenous contrast. It has two major advantages over conventional modalities which identify it as having the potential to fill this role. Firstly, direct visualisation of thrombus overcomes many of the pitfalls of conventional techniques, which have either identified thrombus as a filling defect or in terms of surrogates. Secondly, simultaneous imaging of the legs and chest allows a comprehensive assessment of thrombus load, minimising the importance of overlooked subsegmental pulmonary embolism (PE) and potentially facilitating more titrated treatment. Early data suggest MRDTI is highly accurate for the detection of both deep vein thrombosis (DVT) and PE, and ongoing out-come studies are evaluating the safety of withholding treatment in suspected DVT and PE on the basis of negative MRDTI alone. If favorable, a multi-centre outcome study evaluating cost-effectiveness as well as safety would be justified. Subject to further evaluation, this technique has the potential greatly to simplify and standardise the investigation of suspected VTE. PMID- 12719773 TI - Prevalence of clopidogrel non-responders among patients with stable angina pectoris scheduled for elective coronary stent placement. AB - Dual antiplatelet therapy with aspirin and clopidogrel decreases the rate of stent thrombosis in patients undergoing percutaneous coronary intervention (PCI). However, despite intensified antiplatelet treatment, up to 4.7% of the patients undergoing coronary stenting develop thrombotic stent occlusion, suggesting incomplete platelet inhibition due to clopidogrel resistance. We evaluated the percentage of clopidogrel non-responders among 105 patients with coronary artery disease (CAD) undergoing elective PCI. All patients were treated regularly with aspirin 100 mg/d and received a loading dose of 600 mg clopidogrel followed by a maintenance dose of 75 mg/d before PCI. Clopidogrel non-responders were defined by an inhibition of ADP (5 and 20 Mol/L) induced platelet aggregation that was less than 10% when compared to baseline values 4 h after clopidogrel intake. Semi responders were identified by an inhibition of 10 to 29%. Patients with an inhibition over 30% were regarded as responders. We found that 5 (ADP 5 Mol/L) to 11% (ADP 20 Mol/L) of the patients were non-responders and 9 to 26% were semi responders. Among the group of non-responders there were two incidents of subacute stent thrombosis after PCI. We conclude that a subgroup of patients undergoing PCI does not adequately respond to clopidogrel, which may correspond to the occurrence of thromboischemic complications. Point-of-care testing may help to identify these patients who may then benefit from an alternative antiplatelet therapy. PMID- 12719774 TI - The preparation and phospholipid binding property of the C2 domain of human factor VIII. AB - The C2 domain of human factor VIII was expressed in a yeast secretion system and its binding properties were studied. A cDNA coding the C2 domain sequence of human factor VIII with a N-terminal six amino acids extension (C-C2) was constructed, transformed into Pichia pastoris cells and expressed. The product was purified by ammonium sulfate fractionation and anion exchange chromatography. It emerged as a single peak from both ion exchange and gel filtration columns, indicating C-C2 is a homogenous monomer. The binding activity of C-C2 to phosphatidylserine-containing phospholipid vesicles was measured by competitive binding with annexin V. The values of IC50 were approximately 70nM for both factor VIII and its light chain, but were about 7000nM for C-C2. These results indicated C-C2 has 100-fold less binding affinity than factor VIII or the light chain. Direct binding to solidified phosphatidyl-serine-containing phospholipids also showed that C-C2 has approximately 50-fold less binding affinity than does the light chain. C-C2 poorly inhibited Xase activity. These results together clearly show that the C2 domain alone does not have full membrane binding activity, and suggest that the other light chain domains, A3 and/or C1, are also involved in the phospholipid binding activity of factor VIII. PMID- 12719775 TI - Contributions of Asn2198, Met2199, and Phe2200 in the factor VIII C2 domain to cofactor activity, phospholipid-binding, and von Willebrand factor-binding. AB - The crystal structure of the factor VIII C2 domain consists of a beta-sandwich core from which beta-hairpins and loops extend to form a hydrophobic surface. The hydrophobic surface includes M2199 and F2200 at the tip of the 1(st) beta hairpin. To determine the individual contributions of residues N2198, M2199, and F2200 to phospholipid and von Willebrand factor (vWF) binding properties of factor VIII, we prepared mutant proteins with single alanine substitutions. We found that single mutations at N2198 and M2199 had relatively little impact on cofactor activity, or phospholipid and vWF binding. However the F2200A mutant had slightly lower cofactor activity at subsaturating phospholipid concentrations. Competitive ELISAs suggested that F2200 plays a more important role in both phospholipid-binding and vWF-binding than N2198 and M2199. All mutant proteins were still recognized by a monoclonal antibody and two factor VIII inhibitors that neutralized cofactor activity and blocked factor VIII binding to phospholipids. PMID- 12719776 TI - Effects of recombinant factor VIIa on platelet function and clot structure in blood with deficient prothrombin conversion. AB - While recombinant factor VIIa (rFVIIa) shows promise as a broad-spectrum hemostatic agent, questions remain regarding the most appropriate dose and the best way to monitor its effects. In this study we tested the sensitivity of a thrombin dependent platelet assay, platelet contractile force, to the effects of rFVIIa in normal, factor-deficient, and inhibitor-containing blood samples. Dose dependent effects of rFVIIa on platelet contractile force (PCF) and clot elastic modulus (CEM) were measured in all blood samples. rFVIIa minimally affected PCF and CEM in normal blood clotted with thrombin or batroxobin. While rFVIIa minimally altered PCF and CEM in factor VIII (FVIII) deficient blood clotted with thrombin, rFVIIa increased PCF and CEM and shortened the lag phase in a dose dependent manner in batroxobin-induced clots. The effects of rFVIIa in factor IX (FIX) deficient blood mirrored the effects seen in FVIII deficient samples. Whether clotted with thrombin or batroxobin, baseline PCF and CEM were abnormally low in FVIII deficient samples containing FVIII inhibitors. In such samples, rFVIIa caused dose dependent improvement of PCF, CEM, and lag phases. In one patient with a spontaneous inhibitor, rFVIIa caused dose dependent increases in PCF and CEM in blood clotted with either enzyme. rFVIIa corrects the deficient thrombin generation seen in FVIII and FIX deficiency, and in blood containing FVIII inhibitors. As a consequence, platelet function is improved and clot structure is enhanced. Platelet contractile force and clot elastic modulus measurements are sensitive to the dose dependent effects of rFVIIa. PMID- 12719777 TI - Stimulation of plasminogen activation by recombinant cellular prion protein is conserved in the NH2-terminal fragment PrP23-110. AB - The cellular prion protein (PrP(c)), tissue-type plasminogen activator (t-PA) and plasminogen are expressed in synaptic membranes in vivo. In the central nervous system the fibrinolytic system is associated with excitotoxin-mediated neurotoxicity and Alzheimer's disease. Recently binding of the disease associated isoform of the prion protein (PrP(Sc)) to plasminogen and stimulation of t-PA activity have been reported. In this study the interaction of PrP(c) and plasminogen was investigated using chromogenic assays in vitro. We found that plasmin is able to cleave recombinant PrP(c) at lysine residue 110 generating an NH(2)-terminal truncated molecule that has previously been described as a major product of PrP(c) metabolism. We further characterized the proteolytic fragments with respect to their ability to stimulate plasminogen activation in vitro. Our results show that the NH(2)-terminal part of PrP(c) spanning amino acids 23-110 (PrP23-110) together with low molecular weight heparin stimulates t-PA mediated plasminogen activation in vitro. The apparent rate constant was increased 57 fold in the presence of 800 nM PrP23-110. Furthermore, we compared the stimulation of t-PA activity by PrP(c) and beta-amyloid peptide (1-42). While the activity of the beta-amyloid was independent of low molecular weight heparin, PrP23-110 was approximately 4- and 37 fold more active than beta-amyloid in the absence or presence of low molecular weight heparin. In summary, plasmin cleaves PrP(c) in vitro and the liberated NH(2)-terminal fragment accelerates plasminogen activation. Cleavage of PrP c has previously been reported. Thus cleavage of PrP(c) enhancing plasminogen activation at the cell surface could constitute a regulatory mechanism of pericellular proteolysis. PMID- 12719778 TI - Suppression of argatroban-induced endogenous thrombolysis by PKSI-527, and antibodies to TPA and UPA, evaluated in a rat arterial thrombolysis model. AB - We have previously confirmed, using a rat mesenteric arteriole thrombolysis model, that thrombin inhibition induces endogenous thrombolysis in vivo. In addition, we have shown that thrombin-activatable fibrinolysis inhibitor (TAFI) plays a role in the down regulation of endogenous thrombolysis. However, the mechanism of endogenous thrombolysis or spontaneous plasmin generation in vivo remains unclear. It has been shown in an in vitro system that plasma kallikrein activates pro-urokinase (pro uPA) and/or plasminogen, resulting in plasmin generation. These findings suggest that spontaneous fibrinolysis might be mediated by tPA and plasma kallikrein-dependent uPA. The aim of the present study was to examine whether these mechanisms play a dominant role in endogenous thrombolysis in vivo, using our rat mesenteric arterial thrombolysis model. Argatroban infusion enhanced endogenous thrombolysis. PKSI-527, anti uPA and anti tPA IgGs suppressed argatroban-induced thrombolysis. Also, the antibody IgG preparations suppressed endogenous thrombolysis in the absence of argatroban. In the presence of PKSI-527, anti tPA IgG was more effective than anti uPA IgG in suppressing argatroban-induced thrombolysis. The results suggested that both tPA and plasma kallikrein-mediated uPA activation and tPA release contribute to endogenous fibrinolytic or thrombolytic mechanisms. PMID- 12719779 TI - Anticoagulant serine fibrinogenases from Vipera lebetina venom: structure function relationships. AB - Amino acid sequences of two anticoagulant serine fibrinogenases - alpha- and beta fibrinogenase (VLAF and VLBF) from Vipera lebetina venom have been deduced from the cDNA sequences encoding the enzymes. The mature protein sequences of 234 amino acids (VLAF) and 233 amino acids (VLBF) exhibit significant similarity with other snake venom serine proteinases. Both enzymes contain the catalytic triad His57, Asp102, Ser195, and twelve conserved cysteines forming six disulfide bridges. Unlike typical trypsin-like serine proteinases, they lack the third aspartate, Asp189 which is replaced by Gly189. VLBF is a typical representative of arginine esterases - beta-fibrinogenases. alpha-Fibrinogenase, VLAF, is unique among snake venom serine proteinases with homologous structure. Until now there is no evidence of the anticoagulant serine enzymes degrading fibrinogen alpha chain only and lacking esterolytic activity. PMID- 12719780 TI - Measurement of soluble fibrin monomer-fibrinogen complex in plasmas derived from patients with various underlying clinical situations. AB - We previously reported a monoclonal antibody named IF-43 that specifically recognizes thrombin-modified fibrinogen (desAA- and desAABB- fibrin monomer) bound with fibrinogen or other D(1) domain-containing plasmic fragments such as fragments X,Y, and D(1), but not intact fibrinogen or cross-linked fibrin degradation products (XDP). Here, we tentatively named such complexes, soluble fibrin monomer (FM) -fibrinogen complex. By utilizing IF-43, we have developed a kit to measure soluble FM-fibrinogen complex and compared the profiles with those of two established molecular markers for thrombo-embolic disorders: i.e. the thrombin-antithrombin complex (TAT) and the D-dimer in plasma of patients who underwent surgery without any thrombo-embolic complications. The result indicated that soluble FM-fibrinogen complex is a distinct entity from the two established molecular markers. We have also attempted to observe their profiles in patients with the disseminated intravascular coagulation syndrome (DIC). Although the pro files of soluble FM-fibrinogen complex in individual patients appeared to vary from one patient to the other, the plasma level of soluble FM-fibrinogen complex was found to be increased at the initial phase of disseminated intravascular coagulation syndrome. Thus, the soluble FM-fibrinogen complex may serve as an independent molecular marker for the detection of thrombin generation and the diagnosis of thrombosis. The soluble FM-fibrinogen complex may also serve as a risk factor for thrombosis, because it may precipitate as insoluble complexes beyond its threshold in plasma, or when it is modified by thrombin. PMID- 12719781 TI - The effect of beta-receptor blockade on factor VIII levels and thrombin generation in patients with venous thromboembolism. AB - High factor VIII (FVIII) is a risk factor for venous thromboembolism (VTE). The pathomechanism by which high FVIII leads to an increased risk of VTE is unknown. Physical activity and infusion of adrenalin provoke a rise in FVIII, which can be blocked by a nonselective beta-blockade. We tested the hypothesis that in patients with a VTE beta-blockade decreases FVIII and inhibits coagulation activation. 17 male patients with high FVIII (> 170 IU/dL, n = 7) or low FVIII (<150 IU/dL, n = 10) and a history of VTE received 40 mg of propranolol thrice daily for 14 days. FVIII and vasopressin levels were measured before and during propranolol intake and 28 days thereafter. At the same time points, haemostatic system activation was investigated by measuring prothrombin fragment f1.2 (f1.2) and thrombin antithrombin complexes (TAT) in venous blood and in blood emerging from a skin incision (shed blood). The mean FVIII level before propranolol was 192 IU/dL and 115 IU/dL in patients with high and low FVIII, respectively. During and 28 days after propranolol, no significant change in FVIII was seen in both groups. Changes in f1.2 and TAT were not detectable in either venous blood or in shed blood. beta-receptor blockade did not lower FVIII or inhibit haemostatic system activation in patients with VTE and persistently high FVIII. Administration of propranolol cannot be recommended as secondary thromboprophylaxis in patients with high FVIII. PMID- 12719783 TI - The deletion polymorphism in the angiotensin-converting enzyme gene is a moderate risk factor for venous thromboembolism. AB - ACE displays potent vasoconstrictive effects, attenuation of fibrinolysis, and platelet activation and aggregation, thus possibly promoting venous thromboembolism (VTE). The ACE gene contains an insertion (I) or deletion (D) polymorphism accounting for 50% of the variation in serum ACE concentration. To evaluate the role of the I/D polymorphism in VTE, its prevalence was determined in 931 patients with VTE and 432 blood donors. The prevalence of the DD genotype was 27.6% in patients and 21.3% in controls (OR 1.4; p < 0.02). In multivariate analysis there was a trend of the DD genotype to be an independent risk factor (OR 1.4; p = 0.08). No differences in DD genotype prevalence according to exogenous risk factors were found. Coinheritance of FV G1691A, PT G20210A mutation, and PS deficiency with the DD genotype increased the relative risk of VTE. Thus, the ACE DD genotype is a moderate risk factor of hereditary thrombophilia. Exogenous risk factors did not alter the manifestation of VTE among carriers of the DD genotype, whereas coinheritance of the DD genotype with the aforementioned defects increased the risk for VTE considerably. PMID- 12719782 TI - Clopidogrel and warfarin: absence of interaction in patients receiving long-term anticoagulant therapy for non-valvular atrial fibrillation. AB - The effect of concomitantly administered clopidogrel on anti-coagulation status was investigated in patients receiving long-term warfarin therapy. Forty-three patients with non-valvular atrial fibrillation who were receiving long-term warfarin and had a stable international normalized ratio (INR) between 2 and 3 were randomly assigned to clopidogrel 75 mg daily or placebo for 8 days (Days 1 8). INR (primary endpoint) and plasma levels of warfarin enantiomers (secondary endpoint) were evaluated at Days 3, 6, 9, 13 and 22. Mean INR remained extremely stable in the clopidogrel group, the maximum percentage change from baseline being 0.6% at Day 6. Plasma levels of R- and S-warfarin also remained very stable in those receiving clopidogrel. No serious adverse events, premature discontinuations of study drug or bleeding occurred with clopidogrel. In conclusion, the stable anticoagulation status of patients receiving long-term warfarin therapy is unaffected by concomitant administration of clopidogrel 75 mg daily. PMID- 12719784 TI - Thrombin binding to GPIbalpha induces platelet aggregation and fibrin clot retraction supported by resting alphaIIbbeta3 interaction with polymerized fibrin. AB - We have investigated the mechanisms leading to platelet aggregation following thrombin interaction with the glycoprotein (GP) Ib-IX-V complex. We show that platelets desensitized for the two thrombin receptors, PAR-1 and PAR-4, are still able to aggregate in response to thrombin and that this aggregation can be inhibited by a monoclonal antibody (VM16d) that blocks thrombin binding to GPIbalpha, or by pretreatment of platelets with Mocarhagin, a protease that specifically cleaves GPIbalpha. The thrombin/GPIbalpha-initiated signaling cascade induces platelet shape change through activation of the Rho kinase p160ROCK, independent of calcium mobilization, transient MEK-1 phosphorylation as well as the cleavage of talin through a calcium-independent mechanism. This signaling cascade does not induce the exposure of high affinity alphaIIbbeta3 integrin receptors, nor does it lead to micro -calpain cleavage of filamin or the integrin cytoplasmic tail. In contrast, we provide evidence that binding of thrombin to GPIbalpha induces fibrin binding to resting alphaIIbbeta3 leading to fibrin-dependent platelet aggregation and clot retraction, that can be selectively inhibited by alphaIIbbeta3 antagonists such as RGDS, the dodecapeptide or lamifiban, as well as by the fibrin polymerization inhibitor GPRP-amide. PMID- 12719785 TI - Green tea epigallocatechin-3-gallate inhibits platelet signalling pathways triggered by both proteolytic and non-proteolytic agonists. AB - Epigallocatechin-3-gallate (EGCG), a component of green tea, inhibits human platelet aggregation and cytosolic [Ca(2+)](c) increases more strongly when these processes are induced by thrombin than by the non-proteolytic thrombin receptor activating peptide (TRAP), thromboxane mimetic U46619, or fluoroaluminate. In line with the previously demonstrated EGCG anti-proteolytic activity, a marked inhibition on aggregation is obtained by pre-incubation of thrombin with EGCG prior to addition to cellular suspension. The catechin also reduces cellular Ca(2+) influx following thapsigargin-induced calcium emptying of endoplasmic reticulum, and the agonist-promoted cellular protein tyrosine phosphorylation. Both tyrosine kinases Syk and Lyn, immuno-precipitated from stimulated platelets, are greatly inhibited upon cellular pre-incubation with EGCG, which also inhibits the in vitro auto-phosphorylation and exogenous activity of these two enzymes purified from rat spleen. Both thrombin-induced aggregation and [Ca(2+)](c) increase are reduced in platelets from rats that drank green tea solutions. It is concluded that EGCG inhibits platelet activation, by hindering the thrombin proteolytic activity, and by reducing the agonist-induced [Ca(2+)](c) increase through inhibition of Syk and Lyn activities. PMID- 12719786 TI - Anandamide induces apoptosis in human endothelial cells: its regulation system and clinical implications. AB - Anandamide (AEA), an endogenous cannabinoid, is generated by macrophages during shock conditions, and is thought to be a causative mediator of septic shock. Thus, we hypothesized that AEA plays a crucial role in endothelial cell (EC) injury. Here, we demonstrate that AEA induces apoptosis in a time-and dose dependent manner in human umbilical vein endothelial cells (HUVECs). AEA triggered phosphorylation of c-Jun NH(2)-terminal kinase (JNK) and p38 mitogen activated protein kinase. AEA also showed a marked increase of interleukin Ibeta- converting enzyme (ICE)CED-3 family protease (caspase-3) activity. AEA-induced EC death was inhibited by a selective vanilloid receptor 1 (VR1) antagonist, capsazepine, and was enhanced by a VR1 agonist, capsaicin, indicating that AEA induces apoptosis in ECs via VR1. In conclusion, we propose that AEA may play a crucial role in EC injury under conditions of shock, and that the use of inhibitors of the AEA regulation system may have a therapeutic effect under these conditions. PMID- 12719787 TI - Preoperative plasma fibrinogen levels predict mortality after coronary artery bypass grafting. AB - This study was designed to investigate whether plasma fibrinogen levels as well as the beta-fibrinogen -455 G/A genotype are associated with outcome after coronary artery bypass graft (CABG) operation. We enrolled 249 consecutive CAD patients one day before they underwent a CABG operation. Data from 220 patients with available plasma fibrinogen levels were analyzed. The primary end-point was total mortality, the secondary end-point mortality from cardiac causes or the need for myocardial revascularization. The 2-year total mortality was 9.1% in the entire cohort. Multivariable analysis revealed an independent relationship between the primary end-point and preoperative plasma fibrinogen levels but not the beta-fibrinogen -455 G/A genotype. Neither preoperative plasma fibrinogen levels nor the beta-fibrinogen -455 G/A genotype could predict the secondary end point. We conclude, that elevated preoperative plasma fibrinogen levels, but not the beta-fibrinogen -455 G/A genotype predict the total mortality after CABG operation. PMID- 12719788 TI - Evidence of prolonged disturbances in the haemostatic, hemorheologic and inflammatory profiles in transmural myocardial infarction survivors. AB - Haemostatic, hemorheologic and inflammatory disturbances have been associated with acute coronary syndromes. Most knowledge is reported in cross sectional studies and are without time dependent evolution of these profiles. The aim of this study was to evaluate, during the first year, the evolution of the haemostatic, hemorheologic and inflammatory profiles determined at hospital discharge in survivors with transmural myocardial infarction (MI). Eighty eight (79 male; 9 female) mean age of 58 +/- 11 years, survivors of a transmural MI were prospectively studied at discharge, 6 months and one year after the event. Haemostatic (protein C, antithrombin III and plasminogen activator inhibitor 1), hemorheologic (blood fluidity and components) and inflammatory profiles (polymorphonuclear elastase and leukocyte count) were determined using standard methodology. The results of the study can be summarized as follows: (1) Protein C decreased (p < 0.05) over time while PAI-1 only varied significantly until 6(th) month. (2) Plasma viscosity and fibrinogen (p < 0.001) decrease over time, while erythrocyte aggregation (p < 0.001) and haematocrit increased. Whole blood viscosity did not vary. (3) Leukocyte decreased (p < 0.001) and elastase did not (4). Those patients with cardiovascular events (n = 7) had higher PAI-1 concentration (p < 0.05) and leukocyte count (p < 0.01), at discharge (5) Left ventricle ejection fraction correlated significantly with plasma viscosity (r = 0.35 p < 0.05). The results of this longitudinal study show dynamic modifications of the haemostatic, hemorheologic and inflammatory profiles during the first year of a transmural myocardial infarction. In addition, there are interrelations between them and the clinical profile that could help to explain the clinical evolution of this group of patients. PMID- 12719789 TI - Urokinase-induced migration of human vascular smooth muscle cells requires coupling of the small GTPases RhoA and Rac1 to the Tyk2/PI3-K signalling pathway. AB - Urokinase-type plasminogen activator (uPA) facilitates cell migration by localizing proteolisys on the cell surface and by inducing intracellular signalling pathways. In human vascular smooth muscle cell (VSMC) uPA stimulates migration via the uPA receptor (uPAR) signalling complex containing the Janus kinase Tyk2 and phosphatidylinositol 3-kinase (PI3-K). We report that active GTP bound forms of small GTPases RhoA and Rac1, but not Cdc42, are directly associated with Tyk2 and PI3-K in an uPA/uPAR-dependent fashion. Endogenous RhoA, but not Rac1 or Cdc42, was significantly activated in response to uPA. RhoA activation was abolished by cell treatment with two unrelated, structurally distinct, specific inhibitors of PI3-K, wortmannin, and LY294002. Downstream of RhoA, phosphorylation of myosin light chain (MLC) was dramatically upregulated by uPA in a Rho kinase- and PI3-K-dependent manner. Thus, selective Rho kinase inhibitor Y27632 and PI3-K inhibitors wortmannin and LY294002 prevented the uPA induced stimulation of MLC phosphorylation. Rho kinase inhibition also decreased uPA-stimulated VSMC migration as observed in a Boyden chamber. VSMC immunocytochemical staining demonstrated redistribution of RhoA and Rac1 active forms to the newly formed leading edge of migrating cell. VSMC microinjection with antibodies to either Rho or Rac1 decreased uPA-stimulated cell migration, indicating the involvement of both GTPases in the migration process. Our results provide evidence that the small GTPases RhoA and Rac1, together with Rho kinase, are necessary to mediate the uPA/uPAR-directed migration via the Tyk2/PI3-K signalling complex in human VSMC. PMID- 12719790 TI - Inhibition of PMN-elastase activity by semisynthetic glucan sulfates. AB - Proteolysis of connective tissue by enzymes such as PMN-elastase (PMNE) is a crucial step during inflammation and metastasis. Semisynthetic sulfated carbohydrates (SC) were shown to exhibit potent antiinflammatory and antimetastatic activity in vivo. The aim of the present study was to examine whether interferences with PMN-elastase may contribute to these effects. Therefore, the interactions of these compounds with PMNE were evaluated in various test systems. Besides semisynthetic alpha-1,4/1,6- and beta-1,3-glucan sulfates, UFH, a LMWH and pentosan polysulfate (PPS) were included in the study. The inhibitory activity of SC improves not only with increasing molecular weight (MW 10 - 250 kDa: 37 - 54% inhibition at 0.25 micro g/ml) and degree of sulfation (DS 0.25 - 2.0: 16 - 50% inhibition at 0.25 micro g/ml), but depends also on their genuine polysaccharide structure (IC50 beta-1,3-glucan sulfate 0.18 / alpha 1,4/1,6-glucan sulfate 0.25 / UFH 0.5 micro g/ml). Using physiological substrate assays (collagen, elastin), beta-1,3- and alpha-1,4/1,6-glucan sulfates are more active than UFH (inhibition at 1.5 micro g/ml: 41 / 32 / 12%). According to enzyme-inhibitor binding studies, SC exhibit structure dependent affinity to the enzyme (K(d) for PMNE: beta-1,3 < alpha-1,4/1,6 < UFH). Finally, SC were shown to inhibit cancer cell-mediated elastinolysis. PMID- 12719791 TI - Reactive oxygen species modulate HIF-1 mediated PAI-1 expression: involvement of the GTPase Rac1. AB - The hypoxia-inducible transcription factor HIF-1 mediates upregulation of plasminogen activator inhibitor-1 (PAI-1) expression under hypoxia. Reactive oxygen species (ROS) have also been implicated in PAI-1 gene expression. However, the role of ROS in HIF-1-mediated regulation of PAI-1 is not clear. We therefore investigated the role of the GTPase Rac1 which modulates ROS production in the pathway leading to HIF-1 and PAI-1 induction. Overexpression of constitutively activated (RacG12V) or dominant-negative (RacT17N) Rac1 increased or decreased, respectively, ROS production. In RacG12V-expressing cells, PAI-1 mRNA levels as well as HIF-alpha nuclear presence were reduced under normoxia and hypoxia whereas expression of RacT17N resulted in opposite effects. Treatment with the antioxidant pyr-rolidinedithiocarbamate or coexpression of the redox factor-1 restored HIF-1 and PAI-1 promoter activity in RacG12V-cells. In contrast, NFkappaB activation was enhanced in RacG12V-cells, but abolished by RacT17N. Thus, these findings suggest a mechanism explaining modified fibrinolysis and tissue remodeling in an oxidized environment. PMID- 12719793 TI - Cross-linking of fibrinogen by factor XIII zymogen is not apparent in vivo. PMID- 12719792 TI - Rapid genotyping of haemostatic gene polymorphisms using the 5' nuclease assay. AB - Hemostatic gene polymorphisms have been shown to be associated with arterial and venous thrombotic disease. To date these polymorphisms have mainly been detected by labor-intensive conventional gel based methods. Aim of the present study was to design and optimize high throughput 5' nuclease assays for the detection of a set of 10 single-nucleotide polymorphisms (SNP) in genes of importance for hemostasis: plasminogen activator inhibitor type 1 -675 4G>5G, thrombin activatable fibrinolysis inhibitor Ala147Thr and 1,542C>G, beta-fibrinogen 455G>A, von Willebrand factor -1,051A>G, factor VII Arg353Gln, factor XIII Val34Leu, prothrombin 20,210G>A, tissue factor pathway inhibitor -287T>C, and methylenetetrahydrofolate reductase 1,298A>C. Specificity of each genotyping assay was confirmed by sequence-based typing and reproducibility was evaluated by repeated genotyping. The genotyping protocols presented here may serve as a valuable tool for clinical researchers interested in exploring associations between these SNPs and thrombotic disease. PMID- 12719795 TI - High prevalence of prothrombin G20210A mutation among patients with deep venous thrombosis in Lebanon. PMID- 12719796 TI - Stabilyte tubes that contain strongly acidic citrate prevent in vitro complex formation between activated protein C and protein C inhibitor. PMID- 12719797 TI - The effect of simvastatin co-medication on warfarin anticoagulation response and dose requirements. PMID- 12719798 TI - Combination of microfracture and periostal-flap for the treatment of focal full thickness articular cartilage lesions of the shoulder: a prospective study. AB - Focal full-thickness articular cartilage lesions of the shoulder are less common than those of the lower extremity but are often symptomatic and may progress to degenerative osteoarthritis. This prospective study evaluated our clinical results for cartilage repair in five patients with chondral defects localized at the humeral head using a combination of microfracture and periostal flap, all by deltoidopectoral approach. Mean follow-up was 25.8 months (range 24-31) and consisted of a clinical examination, Constant score examination, radiography, and magnetic resonance imaging; three patients underwent a second-look arthroscopy an average of 8 months following cartilage repair. We found the Constant score significantly improved over the preoperative level, from 43.4% to 81.8%. Pain was reduced significantly to 18.6 points. Radiography and magnetic resonance imaging showed progression of the osteoarthritis in two patients. Second-look arthroscopy revealed a significantly reduced cartilage lesion. This is the first report of a combination of microfracture and a periostal flap for repair of focal full thickness cartilage lesions at the shoulder. Short-term follow-up clinical results were satisfactory. It is essential to address the underlying pathology. Results must be reconfirmed in a long-term study. PMID- 12719800 TI - Portal vein thrombosis: an emergency solution for blood flow in liver transplantation. AB - Recipient portal vein thrombosis in liver transplantation is a contingency that increases surgical difficulty as well as patient morbidity and mortality. The aim of this paper is to demonstrate a surgical technique for reconstruction of portal blood flow in emergency situations of portal vein thrombosis with inadequate blood flow and a poor vascular bed for re-vascularization. PMID- 12719799 TI - Induction of indirect donor-specific hyporesponsiveness by transportal RT1 peptide pulse in rat skin transplantation. AB - In the present study, we examined whether transportal pulse of class I major histocompatibility complex (MHC) allopeptides can induce indirect (non-chimeric) donor-specific hyporesponsiveness, using a high-responder rat skin transplantation model. Two donor-specific 8-amino acid peptides corresponding to residues 58-65 and 70-77 in the alpha(1) helical region of RT1.A(a) were synthesized. In order to test immunogenicity of these peptides, mixed lymphocyte reaction (MLR) was performed. Then, 100-microg portions of peptides were injected into recipient Lewis (LEW, RT1.A(l)) rats via the portal vein 14 days before skin transplantation. Skin allografts from August Copenhagen Irish (ACI, RT1(a)) or Wistar King A (WKA, RT1(k), third-party) donors were transplanted to LEW (RT1(l)) recipients. Transportal pulse of residues 58-65 and 70-77 prolonged graft survival significantly in ACI-to-LEW skin transplantation (17.6+/-0.40 and 18.0+/ 0.45 days) compared with control (14.2+/-0.37 days). However, pulse of residues 106-113, a non-donor-specific control, did not prolong graft survival time (14.6+/-0.40 days) in the same combination. Regarding the third-party donor, residues 58-65 injected into LEW recipients had no effect on survival time of skin grafts (19.0+/-0.84 days) derived from WKA donors compared with the untreated WKA-to-LEW control (19.4+/-0.93 days). Transportal pulse of RT1.A(a) peptides induced donor-specific hyporesponsiveness even in a high-responder rat skin transplantation model. Our results suggest that graft enhancement by transportal exposure to donor cells may not be induced by a chimeric process but, instead, by an indirect mechanism not involving intervention of viable donor cells. PMID- 12719801 TI - Increased expression of p21 (WAF1/CIP1) cyclin-dependent kinase (CDK) inhibitor gene in chronic allograft nephropathy correlates with the number of acute rejection episodes. AB - The p21 (WAF1/CIP1) cyclin-dependent kinase (CDK) inhibitor gene is considered to be the senescence marker in some recent publications. Expression of the gene was evaluated in 14 normal human kidney tissues of different ages and in nine chronically rejected renal allografts. All normal kidneys were negative for p21 expression. Glomerular, tubular and interstitial expression of the marker was detected in 88.9% ( P<0.0001) and vascular expression in 66.7% of chronically rejected grafts ( P<0.001). No correlation was found between the intensity of p21 expression and recipient age, donor age or number of human leukocyte antigen (HLA) mismatches. The marker was expressed more in grade 3 of chronic allograft nephropathy (CAN) than in grade 2 ( P=0.059 for glomerular score). Tubular expression of p21 was correlated with the number of acute rejections: P<0.05 for three vs one and two, and P=0.0046 for three vs no previous acute rejection episodes. PMID- 12719802 TI - Ascorbic acid against reperfusion injury in human renal transplantation. AB - The cadaveric renal graft is exposed to ischaemic injury during preservation and to oxidative damage during reperfusion. Both these mechanisms are known to cause cell damage, which may impair graft function. Reperfusion injury (RPI) is mediated by reactive oxygen species (ROS). Ascorbic acid (AA) is a potent physiological extracellular scavenger of ROS. We perfused 31 renal grafts immediately before implantation with a solution of Euro-Collins containing 0.5 mg/ml of AA to diminish RPI. From every donor, the contralateral kidney served as a control. The control grafts were perfused with the same perfusion as those of the AA group, only without the AA substitution. We assessed the effect of AA by recording serum creatinine, creatinine clearance, initial graft function and early rejections. The incidence of delayed graft function (DGF) was 32% in the AA group, and 29% in the control group. Other parameters were also similar in both groups, except for the length of DGF, which showed a trend towards a shorter duration in the AA group. The pre-operative systemic AA concentration was significantly ( P=0.01) lower in the haemodialysis patients than in those on peritoneal dialysis. In conclusion, this clinical study could not demonstrate significant benefits of AA in renal transplantation. PMID- 12719803 TI - Histamine-degrading enzymes as cellular markers of acute small bowel allograft rejection. AB - Intestinal histamine-degrading enzymes diamine oxidase (DAO) and histamine N methyltransferase (HNMT) activities are relatively constant per individual and bowel segment, and they reflect the functional integrity of the intestinal mucosa. It was, therefore, hypothesised that a decrease in these enzymes could be indicative of acute rejection of an intestinal allograft. Enzymatic activities of DAO and HNMT were determined in mucosal biopsies of isogeneic (Lewis-to-Lewis, n=48) and allogeneic (Brown Norway-to-Lewis, n=48) heterotopic small bowel transplants in a rat model at various time periods. Allograft recipients were not given any immunosuppression. While no changes in enzyme activities were observed in isografts up to day 8 following transplantation, significantly reduced activities of both enzymes were found in all allografts 6-8 days after transplantation. Activities of both DAO and HNMT exhibited a strong negative correlation with the histological rejection score ( P<0.01). We can conclude that DAO and HNMT activities in gut mucosa are reliable quantitative markers of acute intestinal allograft rejection in the rat that support histopathological analysis. PMID- 12719804 TI - ABO-incompatible organ and bone marrow transplantation: current status. AB - Despite the presence of preformed antibodies against AB oligosaccharide epitopes on the donor vascular endothelium, approximately one-third of ABO-incompatible organ allografts are not rejected by a humoral mechanism. With the growing immune manipulation of the recipient, survival rates can be raised considerably, although they remain significantly inferior to those of ABO-compatible transplantation. Data from the Collaborative Transplant Study indicate a 1-year graft survival rate of approximately 50-60% following cadaveric ABO-incompatible kidney, liver or heart transplantation, compared with 70-80% for an ABO compatible organ. The results for infants and young children, however, are very much better than those of adults, particularly for liver and heart transplantations, and the data suggest that B-cell tolerance can develop in the infant age group. We here review clinical and experimental experience with ABO incompatible organ and bone marrow allotransplantation and address the mechanisms by which organs or cells survive in the presence of natural anti-carbohydrate antibodies. PMID- 12719806 TI - Novel plant bioassays for monitoring the genotoxicity of drinking water from the inhabited areas of the Ukraine affected by the Chernobyl accident. PMID- 12719805 TI - Extracorporeal membrane oxygenation as a bridge to cardiac transplantation in a patient with cardiomyopathy and hemophilia A. AB - OBJECTIVE: To report the use of extracorporeal membrane oxygenation (ECMO) as a bridge to cardiac transplantation in a patient with hemophilia A and dilated cardiomyopathy. DESIGN: Case report. INTERVENTIONS AND RESULTS: During ECMO factor VIII concentrate was administered to maintain a factor VIII level close to 50% of normal control both to decrease the risk of bleeding and to allow standard anticoagulation with heparin. The patient developed an inhibitor to exogenous factor VIII during hospitalization, which was successfully managed with a high dose continuous infusion of factor VIII during ECMO, the transplant operation, and the postoperative period. CONCLUSIONS: We report the successful use of ECMO as a bridge to cardiac transplantation in a patient with hemophilia A and low level factor VIII inhibitors. PMID- 12719807 TI - Genotoxicity, catalase, and acetylcholinesterase in the assessment of the pollution status of some sites on the tunisian littoral. PMID- 12719808 TI - Rapid bioassay for microcystin toxicity based on feeding activity of Daphnia. PMID- 12719809 TI - Detection of aircraft deicing additives in groundwater and soil samples from Fairchild Air Force Base, a small to moderate user of deicing fluids. PMID- 12719810 TI - Distribution of 1,4-dioxane and N, N-dimethylformamide in river water from Niigata, Japan. PMID- 12719811 TI - Dissipation, distribution, and uptake of 14C-chlorpyrifos in a model tropical seawater/sediment/fish ecosystem. PMID- 12719812 TI - Pesticide peak discharge from wastewater treatment plants into streams during the main period of insecticide application: ecotoxicological evaluation in comparison to runoff. PMID- 12719813 TI - Effect of nonylphenol on apoptosis of Sertoli cells in vitro. PMID- 12719814 TI - Copper release from copper tableware. PMID- 12719815 TI - Polychlorinated biphenyls and organochlorine pesticides in human milk from the locality Prague, Czech Republic: a comparative study. PMID- 12719816 TI - Differences in persistent organochlorine pesticides concentration between breast adipose tissue and blood serum. PMID- 12719817 TI - In vitro bioaccessibility of metals in soils from a superfund site in Puerto Rico. PMID- 12719818 TI - Metal contamination of soils in the Shenyang Zhangshi irrigation area. PMID- 12719819 TI - Arsenic, cadmium, chromium, copper, lead, and zinc contamination in mine tailings and nearby streams of three abandoned mines from Korea. PMID- 12719820 TI - Metals in the green sea urchin (Strongylocentrotus droebrachiensis) as an indicator for the near-field effects of chemical wastes from salmon aquaculture sites in New Brunswick, Canada. PMID- 12719822 TI - Polycyclic aromatic hydrocarbons in sediments from Kolleru wetland in India. PMID- 12719821 TI - Analysis of airborne volatile organic compounds of a steel industry by solid phase microextraction and gas chromatography/mass spectrometry. PMID- 12719823 TI - Polycyclic aromatic hydrocarbons in soil adjacent to highways in Beijing, People's Republic of China. PMID- 12719824 TI - Sedimentary nonylphenol contamination in an urbanized, industrialized segment of the Delaware River estuary, USA. PMID- 12719825 TI - Toxicity of Chlorpyrifos to the fish Oreochromis mossambicus. PMID- 12719826 TI - Effect of sub-lethal concentrations of endosulfan on hematological and serum biochemical parameters in the carp Cyprinus carpio. PMID- 12719827 TI - Effect of beta-cyclodextrin compounds on the solubilization of three selected pesticides and their toxicity with methyl parathion to Rana tigrina tadpoles. PMID- 12719828 TI - Short-term toxicity of ammonia, nitrite, and nitrate to the aquatic snail Potamopyrgus antipodarum (Hydrobiidae, Mollusca). PMID- 12719829 TI - Bioaccumulation of pesticides on some organs of freshwater catfish Mystus vittatus. PMID- 12719830 TI - Acute toxicity of organic and inorganic compounds on the freshwater cyclopoid copepod Eucyclops neumani neumani (Pesta, 1927). PMID- 12719831 TI - Zinc effects on the embryos and larvae of the sharptooth catfish, Clarias gariepinus (Burchell, 1822). PMID- 12719832 TI - Intake and assimilation of zinc, copper, and cadmium in the terrestrial isopod Porcellio scaber Latr. (Crustacea, Isopoda). PMID- 12719833 TI - Effects of zinc upon tolerance and heavy metal uptake in alfalfa plants (Medicago sativa). PMID- 12719834 TI - Effects of hexavalent chromium in two strains of Euglena gracilis. PMID- 12719835 TI - Survival and growth of diazotrophic cyanobacterial isolates exposed to rice-field herbicides. PMID- 12719836 TI - Effect of HCH and fenvalerate on growth and distribution of microorganisms in relation to persistence of the insecticides in the rhizosphere soils of wetland rice. PMID- 12719837 TI - Social inequalities and the common mental disorders: a systematic review of the evidence. AB - BACKGROUND: Of two large-scale government-commissioned studies of common mental disorders in the UK, one found occupational social class to be the strongest marker of risk while the other showed no clear relationship. This study reviews the published evidence on the links between conventional markers of social position and the common mental disorders in developed countries. METHODS: Inclusion criteria covered general population based studies with broad social class variation; samples of 3,000 or more adults of working age; identification of mental illness by validated instruments; social position identified by explicit standard markers; fieldwork undertaken since 1980; published output on key areas of interest. Incompatible study methods and concepts made statistical pooling of results invalid. RESULTS: Of nine studies, eight provide evidence of an association between one or more markers of less privileged social position and higher prevalence of common mental disorders. For some individual indicators in particular studies, no clear trend was evident, but no study showed a contrary trend for any indicator. The more consistent associations were with unemployment, less education and low income or material standard of living. Occupational social class was the least consistent marker. CONCLUSIONS: Common mental disorders are significantly more frequent in socially disadvantaged populations. More precise indicators of education, employment and material circumstances are better markers of increased rates than occupational social class. PMID- 12719838 TI - Social position and the common mental disorders with disability: estimates from the National Psychiatric Survey of Great Britain. AB - BACKGROUND: Published studies linking the common mental disorders with social disadvantage lack basic comparability. This project aimed to estimate effect sizes and independence of social position markers as risk factors for common mental disorders. Disorders with disability were examined to identify groups with high clinical and policy priority. METHODS: Data from the 1993 household survey of psychiatric morbidity in Great Britain were analysed using logistic regression models, using traditional and more specific markers of social position. RESULTS: Of those with a common mental disorder, 22 % reported difficulty doing at least one activity of daily living, linked to their mental symptoms. In comprehensive statistical analyses, having two or more physical illnesses was associated with an odds ratio of 6.42 (95 % CI 4.34-9.51) for common mental disorder with disability, while odds ratios of 3 or more were present for being economically inactive or having had two or more recent adverse life events. Occupational social class was not an independent marker of raised rates of disorder. Similar patterns of result were present for common mental disorders irrespective of disability, although odds ratios were smaller. CONCLUSIONS: Several specific markers of less privileged status are independently associated with raised rates of common mental disorders, with or without disability. There may be scope to target specific high-risk groups within comprehensive programmes to reduce mental health inequalities. PMID- 12719839 TI - Self-reported happiness in life and suicide in ensuing 20 years. AB - OBJECTIVE: The aim of this study was to investigate the relationship between self reported unhappiness and suicide. METHODS: A sample of adult Finns unselected for health status (N = 29,067) responded to a question on happiness in life with five alternatives: very happy (15.9 %), fairly happy (63.4 %), cannot say (12.9 %), fairly unhappy (6.9 %), very unhappy (1 %). Cox regression was used to study the suicide risk. RESULTS: From 1976 to 1995, 182 suicides occurred. At baseline, unhappiness was associated with older age, male gender, sickness, living alone, smoking, heavy alcohol consumption, physical inactivity and belonging to intermediate social class. The risk of suicide increased with decreasing happiness. The very unhappy compared with the very happy had an age-adjusted hazard ratio (HR) of 10.84 (95 % CI 3.93-29.94) for suicide. After adjusting for the above-mentioned factors this HR was 7.01 (95% CI 2.44-20.08). CONCLUSION: Self-reported unhappiness is strongly associated with subsequent suicide in a long follow-up. Moreover, it seems to indicate also cumulative health hazards. PMID- 12719840 TI - Well-being and health behaviour by parental socioeconomic status: a follow-up study of adolescents aged 16 until age 32 years. AB - BACKGROUND: The aim of the study was to investigate the impact of parental socioeconomic status (SES) on subjects' well-being and health behaviour in adolescence, early adulthood and adulthood, and whether these impacts remained after controlling for the person's own SES. METHODS: All 16-year-old ninth-grade school pupils of one Finnish city completed questionnaires at school. Subjects were followed up using postal questionnaires when aged 22 and 32 years. RESULTS: Females of manual class origin had lower self-esteem and more distress symptoms from adolescence to adulthood than those from a non-manual background. Lower self esteem was found among males from manual class families in adolescence and early adulthood. In both genders, no significant class differences were found in depression, health status or prevalence of chronic illness. Unhealthier behaviours regarding smoking and physical activity were more prevalent among both genders of manual class origin, and females of this group had higher rates of overweight and higher body mass index scores. After controlling for the person's own SES, the effect of parental SES diminished but remained significant for smoking in both genders and for physical activity in males up to 22 years, and for self-esteem and BMI in females up to 32. CONCLUSIONS: This follow-up study contributes to the health inequality debate investigating parental SES differences in health behaviour and somatic health, and particularly in psychological health, which is relatively rarely investigated. The results indicate that parental SES has effects on early adult and adult well-being and health behaviour other than those mediated by current SES. PMID- 12719841 TI - Psychometric properties of an index of emotional distress in the U.S. National Comorbidity Survey. AB - BACKGROUND: The National Comorbidity Survey (NCS; Kessler et al. 1994) was a nationwide household survey of the U.S. population designed to produce data on the prevalence and correlates of psychiatric disorders. The NCS dataset is now in public-use format and continues to be widely used for ongoing research efforts. The NCS dataset included a set of 14 items that have face validity as a measure of current emotional distress (depression and anxiety) and could serve as a potentially useful continuous measure of psychological distress. However, there have been no published studies on its psychometric properties and this measure has not yet been utilized by researchers using the NCS dataset. This paper provides an evaluation of the psychometric properties of the NCS Distress Index. METHOD: The NCS Part II public-use dataset (N = 5877) was used. Detailed diagnostic information was collected along with 14 items assessing current psychological distress and measures of Neuroticism and Openness to Experience. RESULTS: The NCS Distress Index was found to be internally consistent (Alpha = 0.92) and a series of principal-components analyses demonstrated that the measure is most accurately conceptualized as a single-factor measure of general distress. The construct validity of the Distress Index was supported by its associations with the measures of Neuroticism and Openness to Experience. A series of comparisons between diagnostic groups also supported the construct validity of the measure. For example, those with disorders characterized by depressed mood and worry scored higher on the Distress Index than those with disorders characterized by fear and hyperarousal. CONCLUSIONS: The NCS Distress Index is a psychometrically sound measure of current emotional distress. Future studies utilizing the NCS public-use dataset could potentially benefit from the inclusion of this measure in addition to more commonly investigated categorical variables such as diagnosable disorders. PMID- 12719842 TI - Can cognitive deficits explain differential sensitivity to life events in psychosis? AB - BACKGROUND: Life events (LE) have been found to influence the onset and course of psychotic disorders. It has been suggested that LE have their effect by increasing underlying sensitivity to daily life stress, a reported vulnerability marker for psychosis. As increased stress-sensitivity and cognitive impairments have been shown to be negatively associated with each other in patient populations, it is attractive to hypothesise that the impact of LE on sensitivity to daily stress is modified by the degree of cognitive impairment, higher levels of cognitive impairment giving rise to reduced impact of LE on daily life stress sensitivity. METHODS: Patients with psychotic illness (n = 42) were studied with a) a standard battery of neuropsychological tests to assess cognitive functioning, b) the Experience Sampling Method (a structured diary technique assessing current context and mood in daily life) to assess (i) appraised subjective stress related to daily events and activities, and (ii) emotional reactivity conceptualised as changes in both negative affect (NA) and positive affect (PA), and c) the Brown and Harris Life Event and Difficulties Schedule to assess LE over the last year. RESULTS: Multilevel regression analyses showed that a prior history of LE increased the sensitivity for daily life stress in subjects with the best performance on the cognitive tests. CONCLUSIONS: Subjects with cognitive impairments, who already were shown to have lower levels of sensitivity to daily life stress (Myin Germeys et al. 2002), may additionally be less reactive to prior exposure to LE. This result fits with the notion of separate affective and cognitive pathways of symptom formation in psychosis, the cognitive pathway being characterised by severe cognitive deficits and the affective pathway by increased levels of stress-sensitivity associated with prior exposure to LE. PMID- 12719844 TI - From research to practice: a local mental health services needs assessment. AB - BACKGROUND: Service planning based on aggregated needs assessment is rarely practised. There is a lack of research into how population-based needs assessment approaches can be translated into routine clinical settings. AIMS: This study aimed to ascertain the 2-month period prevalence of psychosis in Gloucester City, to investigate the level of need among those in contact with mental health services, and to identify the compromises which are involved in routine collection of data to inform services, compared with specifically funded research studies. Method People with ICD-10 diagnosis of functional psychosis were identified by searching General Practice and mental health service records in Gloucester City. The needs of the sub-sample in touch with mental health services were assessed using the Camberwell Assessment of Need Short Appraisal Schedule (CANSAS). RESULTS: Four hundred and seventy-four cases were identified, including 403 in contact with mental health services. Staff CANSAS data were collected for 225 patients, with a mean rating of 7.0 met and 3.6 unmet needs per patient. Unmet need was higher in the non-Caucasian group. CONCLUSIONS: In this locality, patients with functional psychosis were largely in contact with mental health services, were in employment, were disproportionately looked after by a few City centre General Practitioners, and high levels were in supported accommodation. Higher levels of need were found than previously demonstrated. Systematic assessment of needs with research instruments to contribute to locality service planning is possible without a major research grant, but involves compromises on established research designs. PMID- 12719843 TI - How psychiatric patients perceive the public's stereotype of mental illness. AB - BACKGROUND: It is well established that the general public has devaluating attitudes towards psychiatric patients. In order to avoid rejection, many of these patients develop coping strategies, such as withdrawal and concealing their treatment history. These efforts are in themselves stressing, which might have negative consequences for the course of the disorder. It is not clear, however, how many and which patients do actually perceive the public's stereotype as threatening and, therefore, expect rejection. METHOD: Ninety psychiatric patients and a sample of 1042 persons of the Austrian general population were asked whether they agreed with five devaluating statements about mental patients contained in a questionnaire developed by Link et al. Matched pairs comparisons and multiple logistic regression were employed in order to find out whether patients agreed with these statements to the same extent as the general population did. RESULTS: For the statements that most people believe that psychiatric patients are "less intelligent", "less trustworthy" and "taken less seriously", patients thought significantly less often than the general population that most people devalue mental patients. For two statements ("personal failure", "think less of") no difference was found. CONCLUSIONS: It seems that some psychiatric patients are less convinced than the general population that most people devalue psychiatric patients in specific respects; these patients might fear rejection less than other patients do. Those who actually fear rejection might need antistigma assistance more urgently than the first group. PMID- 12719845 TI - [Surgical principles in haemophilia patients]. AB - Close cooperation between hematologists, surgeons, rehabilitation physicians, pediatricians, psychologists, physiotherapists and nurses is essential for the satisfactory result of any orthopedic surgical procedure in persons with hemophilia. There is no doubt that continuous prophylaxis is the way to avoid orthopedic problems of hemophilia that can still be seen today. Until such a goal can be reached, the orthopedic surgeon will continue performing arthrocentheses, synoviortheses, synovectomies, tendon lengthening, debridements, osteotomies, joint replacements, osteosynthesis of fractures and other less frequent surgical procedures to persons with hemophilia. Over the last 30 years some important achievements have been made in the treatment of hemophilia from the orthopedic point of view. However,some others are still to be solved. The common aim of all of them is to improve the quality of life of persons with hemophilia. PMID- 12719846 TI - [Results after palmar plate-osteosynthesis with angularly stable T-plate in 100 distal radius fractures: a prospective study]. AB - In a prospective study a total of 124 unstable fractures of the distal radius were treated with a fixed angle palmar T-plate (Synthes Ltd., Switzerland). A total of 100 distal radius fractures were evaluated radiologically and clinically after a mean of 10 months after surgery. Loss of correction between postoperative and follow-up radiographs was assessed. The mean loss of the initial volar tilt was 3+/-4 degrees, the loss of radial inclination 0.4+/-2 degrees. The radial shortening was 1+/-1 mm. Flexion and extension of the injured wrist had recovered to an average of 81% of the normal, contralateral side. Radial and ulnar deviation were limited to 84%, pronation and supination reached 91%. Mass grip strength recovered to an average of 74% of the normal side. Using the functional outcome score according to Sarmiento,we obtained 15% excellent,56% good, 28% fair and 1% poor results. The complication rate was 10%. In conclusion, the operative treatment of unstable extra- and intraarticular fractures of the distal radius by fixed angle T-plateosteosynthesis shows good radiological and functional results. PMID- 12719847 TI - [Treatment of trochanteric fractures with the gamma nail]. AB - The osteosynthesis of trochanteric fractures requires a minimal invasive technique to guarantee fully weight bearing and early rehabilitation in elderly patients. In this retrospective study we want to prove if the gamma nail meets all requirements and if the results improved after initiation of the new instrumentarium and standardization of the operative technique. From January 1992 until April 2000 we treated 387 patients with a gamma nail. The average age was 79.4 years. We used the new instrumentarium suitable for radiography since June 1996, the operative technique was standardized since the same time. The evaluation occurred on the basis of clinical and radiological findings included analysis of complications. We've seen intraoperative problems in 29% before June 1996, this rate could be reduced to 9%. General postoperative complications presented in equal frequency. After improvement of the technique dislocations of the neck screw and relevant sinking of the fractures were more rarely. The rate of reoperations after complications of the implant was 8% in both groups. In consideration of technical standards the postoperative full weight bearing and early mobilisation is possible even in old patients. PMID- 12719848 TI - [Influence of nocturnal surgery on mortality and complications in patients with hip fractures]. AB - SUBJECT OF INVESTIGATION: The appropriate time of day for surgery of hip fractures and the question of whether surgery should be performed at night are controversial. We therefore investigated the influence of the time of day on mortality and complication rates in surgery for hip fractures. METHOD: A total of 170 persons were included in the study with 128 operations being performed during the day and 42 at night, after 9 p.m. All patients were operated as early as possible. Patients were randomly assigned to groups. The recorded data were mainly demographic, perioperative and those concerning the patient's history. Mortality and complication rates 6 months after surgery were compared. RESULTS: The two groups were largely homogeneous. Patients operated on at night had a non significantly higher mortality rate. Mortality was highest in those operated on between 9 and 10 p.m. Of those who underwent surgery after midnight, no patient died. No differences were registered with regard to complications. The team of surgeons and anesthetists was equally qualified in both groups. CONCLUSIONS: Given a medical team with equal qualifications and size, we consider nocturnal surgery for hip fractures to be as appropriate as surgery during the day. PMID- 12719849 TI - [Clinical trials in orthopedic and trauma surgery: randomized or non randomized?]. AB - Drawing inferences on a causal relationship between a particular intervention and the observed outcome requires to conduct a clinical experiment which controls for study conditions and systematic errors (bias). This is best to be achieved by randomization in which known and unknown biological risk factors are distributed equally among treatment arms. Trauma and orthopedic surgery,however, occupies an exceptional position in clinical medicine. Random allocation of subjects is often considered difficult because of the tight time frame between patient presentation and the urgent need for surgical treatment, and the dependence of operative results upon technical skills. Evidence of a true treatment effect does not only depend on design issues (i.e., randomized or non-randomized treatment assignment), but on both the prior probability of efficacy and the observed effect size as well. Even though our knowledge of the efficacy of osteosynthesis comparing with, let's say, plaster immobilization or (fictive) placebo therapy is hardly supported by randomized trials, the biologically plausible principle of stable operative fixation of fracture fragments has established itself as the scientific basis to propagate surgical rather than other treatment options. Thus, the efficacy of a medical intervention can be well demonstrated without randomization. Regarding the ultimate goals of stabilization, pain removal, and mobilization,osteosynthesis of a pertrochanteric fracture fits these principles in terms of an all-or-none effect (so called level Ic evidence): without the intervention, effects will not be observed. On the other hand, endpoints such as healing and infection rates or duration of rehabilitation may be severely influenced by confounding factors (e.g., concomitant diseases, age, or gender). Under these circumstances,the goal of quantifying treatment effects of different interventions (i.e., interlocking nails, plates, K-wires) and of discriminating these effects from bias might be solved more reliably by a randomized than by a non-randomized trial.Obviously,the need for randomization relies on the choice of the main endpoint of interest.The postulated overestimation of treatment effects by nonrandomized trials has been proven only for methodologically weak investigations. In contrast, high quality studies led to comparable findings regardless of randomization. In conclusion, there are thinkable alternative designs to randomized trials in trauma surgery, accounting for selected clinical questions and objectives. It must be emphasized that these designs will require a similarly rigorous planning (i.e., study protocols, ethics, sample size considerations) and analysis of the results. PMID- 12719850 TI - [Fracture dislocation of the elbow involving the coronoid process]. AB - INTRODUCTION: Elbow dislocations are associated with osseus lesions in 30-50%. Integrity of the coronoid process is essential for stability of the elbow joint. METHODS: A retrospective study of 39 patients out of 51 was conducted to evaluate a result of surgical treatment in fracture dislocation of the elbow involving the coronoid process. The patients were followed for an average of 45 months. RESULTS: Operative results were assessed using the Morrey-Score. 3 patients presented an excellent, 19 a good, 14 a moderate and 3 a non satisfactory result. CONCLUSION: Results of operative treatment of fracture dislocation of the elbow are essentially determined by the extent of associated osseus lesions of the radial head and the olecranon. To achieve acceptable functional results early reconstruction and fixation of the coronoid process as well as early mobilisation of the joint is necessary. PMID- 12719851 TI - [Occupational disease 2108. Degenerative changes of the cervical spine as a causality criterion in the assessment of discogenic diseases according to BeKV 2108]. AB - With the second amendment to the Ordinance on Occupational Diseases (BeKV) of 18 December 1992, discogenic diseases of the spine are included in the disease register of occupational diseases for the first time. If occupations that impose stress on the spine have been practised for many years, the possibility exists of recognizing degenerative diseases as an occupational disease. In assessment practice, the radiological data on the spine exposed to stress is compared with that of regions which are remote from the stress (cervical/thoracic spine). This pattern of the distribution of degenerative disease is then used as the basis for determining a causal relationship between the occupation causing the stress and disease of the axial skeleton. The pattern of degeneration of the cervical spine was investigated in two groups, one with ( n =153) and one without ( n =333) occupations that impose stress on the lumbar spine. A cumulative score of degenerative changes was elaborated and presented as a new classification. No differences were found between the groups with regard to either the frequency of occurrence, segmental distribution or severity of disease. In both groups, degenerative changes correlated with age. The prevailing assessment practice is discussed on the basis of these data. PMID- 12719852 TI - [Quality of life after calcaneal fractures. A matched-pairs trial with a standardised German control group]. AB - The purpose of the study was the assessment of the limitations of health related quality of life after open reduction and internal fixation (ORIF) of calcaneal fractures and the comparison of these results with those of normal foot-scores. The quality of life of 55 patients with calcaneal fractures treated by ORIF was compared 2.9 years after injury with that of 55 people from the standardised German population of the German-Health-Survey 1998 using the Short Form 36 (SF 36) (Wilcoxon test). The results of SF-36 of patients were correlated with results of the calcaneus score according to Zwipp and AOFAS Ankle Hindfoot Scale (Spearman's correlation coefficient). Health related quality of life showed significant limitations in patients with calcaneal fractures in 9 of 10 scales of the SF-36. Correlations between SF-36 and foot-scores were strong in the function and pain scale and moderate in all other scales. The quality of life of shows significant limitations after calcaneal fractures. Usual foot-scores measure only the function and pain dimension of the outcome. To measure all dimensions, application of the SF-36 can be recommended. International comparisons of results and comparison with a standardised German population is possible. PMID- 12719853 TI - [Arthroscopy of the talocrural articulation]. AB - Arthroscopy of the ankle joint meanwhile enjoys widespread use in those departments that perform arthroscopy so that the application of arthroscopic techniques can be considered a standard procedure also for the talocrural articulation. Various indications have become generally accepted and further areas of application will emerge. As is the case for the knee joint, arthroscopy of the ankle joint solely for diagnostic purposes has become obsolete and due to improved preoperative methods for diagnosis is no longer necessary as the sole procedure. The diagnostic work-up is normally followed by arthroscopic or open treatment. The multitude of clinical examinations and imaging techniques currently available enables noninvasive clarification of pathological conditions in the ankle joint to the greatest possible extent, making detailed preoperative planning possible in most cases. The present article describes the technique, indications, complications, and perspectives for arthroscopy of the talocrural articulation. PMID- 12719854 TI - [Spondylitis due to Salmonella typhimurium]. AB - Very little is known about Salmonella typhimurium as an agent of spondylitis. Only single cases have been described in the international literature over the last years. We report on three patients suffering from spondylitis with Salmonella typhimurium being isolated as the triggering agent and point out the subtly differentiated diagnostic and therapeutic procedures, especially the possible complications of a Salmonella spondylitis. For one of the patients, we diagnosed a concomitant abdominal aortic aneurysm. Another patient sustained an infection of a known aortic aneurysm, which had been operated on 3 years ago. The source of the infection could be either the aneurysm or the spine, with the other structure being infected subsequently. We also discuss possible pathogenesis. PMID- 12719855 TI - [Scapho-capitate fracture syndrome of both hands--first description of a bilateral occurrence of a rare carpal injury]. AB - The scapho-capitate fracture syndrome is a rare but severe injury of the carpus. It is characterized by a rotation of the proximal fragment of the capitate bone of 90 or 180 degrees and a fracture of the navicular bone in the intermediate third. This injury is caused by a fall on the outstretched and dorsiflected hand. The scapho-capitate fracture syndrome was firstly described in 1956 by Fenton. There are 31 cases described in literature but none of these involves both hands. Here we describe the first case of scapho-capitate fracture syndrome in both hands, in a 21 year old woman who jumped out of a window with suicidal intentions. She was treated operatively by Herbert-screws and K-wires. PMID- 12719856 TI - [Avalanche emergency and accidental hypothermia]. AB - The importance of emergency medical treatment for avalanche victims in the pre clinical and clinical sector is still real. Based on new investigations, guidelines for triage have been endorsed by the International Commission of the Alpine Rescue Services (ICAR) to reduce secondary deaths following the successful extrication of victims from the avalanche mass. Although hypothermia plays a secondary role in the total mortality of avalanche victims, the most important task if extrication lasts 35 min or more is the professional treatment of hypothermia. Avalanche emergencies in the southern part of the Black Forest are quite rare. In February 2002 one avalanche victim in this region survived despite a statistically bad prognosis. Based on the current literature, we provide an algorithm which conforms with the ICAR guidelines for emergency personnel and describe the possibilities and standard stage dependent treatment in cases of accidental hypothermia. PMID- 12719857 TI - [The development of a model to calculate the cost of care for the severely injured--an initiative of the Trauma Register of the DGU]. PMID- 12719859 TI - [Development and pre-clinical aspects of pimecrolimus]. AB - Pimecrolimus (SDZ ASM 981), an ascomycin derivative, inhibits the phosphatase calcineurin and blocks the production of inflammatory cytokines in T cells. In contrast to corticosteroids, pimecrolimus has a cell selective mode of action, exerting e.g. no effect on dendritic cells, which have a central function in the skin-associated immune system. Pimecrolimus shows less permeation through skin than corticosteroids and tacrolimus which indicates a lower potential for systemic side effects after topical application. In animal models pimecrolimus has a marked dose-dependent anti-inflammatory activity. However, treatment with pimecrolimus does not induce skin atrophy in contrast to corticosteroids. In contrast to tacrolimus, pimecrolimus does not impair the primary immune reaction in the sensitization phase of allergic contact dermatitis and has generally less effect on systemic immune reactions. In summary, the pharmacological profile of pimecrolimus suggests high clinical efficacy together with excellent safety. PMID- 12719858 TI - Chemokine receptor (CCR2) genotype is associated with myocardial infarction and heart failure in patients under 65 years of age. AB - Inflammation is associated with atherosclerosis of coronary arteries. Chemokines have an important role in inflammation. The CCR2 chemokine receptor mediates leukocyte chemoattraction, which is involved in the pathogenesis of coronary heart disease. We prospectively studied 1960 consecutive patients aged under 65 years and referred for a first-time left ventricular catheter. Left heart catheters were analyzed by two independent cardiologists for the presence of myocardial infarction (regional wall motion abnormality) and moderate or severely reduced left ventricular function on cineventriculography and presence of coronary atherosclerosis on angiography. Genotyping for CCR2 V64I polymorphism was performed. The presence of the rare allele of the CCR2 gene was significantly associated with a higher prevalence of myocardial infarction on cinventriculography (32.0% vs. 24.2%, moderately or severely reduced left ventricular function (14.0% vs. 9.5%) and NYHA class III or IV (16.7% vs. 12.2%). The association of the CCR2 genotype with heart failure was not independent of the presence of myocardial infarction in multivariate analysis. There was no association of the CCR2 genotype with coronary atherosclerosis. The CCR2 genotype seems to predispose patients for myocardial infarction before the age of 65 years. The higher prevalence of heart failure in gene carriers with the rare alle might be a consequence of myocardial infarction. If the CCR2 genotype is associated with higher mortality in the general population must be investigated in further studies. PMID- 12719860 TI - [Antipruritic effects of pimecrolimus and tacrolimus]. AB - The development of topical calcineurin inhibitors resulted in a significant improvement in the treatment of inflammatory skin diseases such as atopic dermatitis. In addition, an excellent amelioration of pruritus could be observed. Other itchy dermatoses such as chronic irritative hand dermatitis, rosacea, graft versus-host-disease, renal pruritus, lichen sclerosus, prurigo simplex, prurigo nodularis, scrotal eczema, and inverse psoriasis also have been treated successfully with pimecrolimus and tacrolimus. The antipruritic effect currently is believed to be related to the inhibition of inflammatory cytokines. Furthermore, recent investigations indicate a release of neuropeptides from sensory nerve fibers and degranulation of mast cells mediated by pimecrolimus and tacrolimus. Similar effects have been observed during capsaicin treatment. These findings may provide a possible explanation for initially observed calcineurin inhibitors related side-effects such as burning and pruritus. Moreover, the antipruritic potency may be related to a direct effect on nerve fibers leading to suppression of itch mediated by unknown mechanisms. PMID- 12719862 TI - [The treatment of atopic dermatitis in adults with topical calcineurin inhibitors]. AB - In about 60% of cases, atopic eczema can persist in adulthood with distinctive clinical features and disease course. The introduction of the topical calcineurin inhibitors pimecrolimus 1% cream and tacrolimus 0.03 and 0.1% ointment clearly improves the long-term management of atopic eczema in adult patients; this has been shown in several large clinical studies and is confirmed by the growing practical experience with these substances. Topical calcineurin inhibitors are, even when applied for weeks and months, safe, well tolerated and efficient; they have a rapid and positive effect on pruritus and the potential--as shown in clinical studies with pimecrolimus 1% cream--to reduce the number of eczema flares, to significantly prolong the time to a first flare and to reduce or even eliminate the need for topical corticosteroids. PMID- 12719861 TI - [Long term management of childhood atopic dermatitis with calcineurin inhibitors]. AB - The calcineurin inhibitors pimecrolimus and tacrolimus are important tools for long-term management of atopic dermatitis in childhood. These drugs are approved in Germany for the intermittent long-term treatment of moderate to severe (tacrolimus ointment) or mild to moderate (pimecrolimus cream) atopic dermatitis in children older than 2 years. Recent study results show that these agents are also effective in infants (aged 3-23 months) with atopic dermatitis. During the long-term treatment of children and infants with calcineurin inhibitors, the therapeutic success can be maintained and there is a continuous increase in the response rate. Long-term use of calcineurin inhibitors in atopic dermatitis is well tolerated locally and systemically, leads to reduction of disease flares and has a corticosteroid sparing effect. PMID- 12719863 TI - [Practice experience with topical calcineurin inhibitors]. AB - The clinical experience with topical calcineurin inhibitors in the field is still rudimentary, if compared to our knowledge of topical corticosteroids. This is especially true for the long-term effects. The systemic transcutaneous absorption that occurs during the therapy of atopic dermatitis is quantitatively irrelevant. However, in the setting of permanent disruption of the skin barrier, as may be encountered in rare Genodermatoses, but as well in cases that do not respond to therapy, continued treatment my result in clinically relevant blood levels. Side effects of topical calcineurin inhibitors may be separated into two groups, i.e. local intolerance reactions, and skin infections. While the typical burning sensation of the newly treated skin is ephemeral, local alcohol intolerance, albeit less frequent, will persist throughout the treatment period. Regarding skin infections, Eczema herpeticatum seems to be the only serious complication; adequate preventive will further reduce the risk of this rare complication. PMID- 12719864 TI - [Sentinel lymph node excision (SLNE) and positron emission tomography in the staging of stage I-II melanoma patients]. AB - BACKGROUND AND OBJECTIVE: Sentinel lymph node excision (SLNE) and positron emission tomography (PET) were evaluated in the staging of 51 Stage I and II melanoma patients (staged according to the guidelines of the German Dermatological Society). PATIENTS/METHODS AND RESULTS: Tumor thickness ranged from 1.0 mm to 6.0 mm (median: 1.5 mm; mean: 2.07 mm). At least one sentinel lymph node (SLN) was excised in all patients; 80 SLN were excised from 69 lymphatic drainage areas. Positive SLN were detected in 6 patients (11.8%). Additional positive lymph nodes were not detected in any of these patients in the following complete lymph node dissection of the affected lymph node basin. Preoperative PET was performed in 40 patients and did not detect any of the micrometastases that were subsequently found by SLNE. During the follow up of 7 40 months (mean 21.9 months) 3 patients experienced tumor progression; 2 of 3 had a positive SLN. CONCLUSIONS: According to the current literature SLNE is recommended in primary tumors greater than 1 mm thickness. PET cannot be expected to give additional information in the staging of stage I-II patients. PMID- 12719865 TI - [Pansclerotic porphyria cutanea tarda after chronic exposure to organic solvents]. AB - A 63 year old man developed generalized scleroderma with massive sclerotic areas, particularly in the abdominal region, four years after being diagnosed with porphyria cutanea tarda (PCT). He had almost daily exposure to organic solvents (benzene, trichlorethylene) for many years. The cutaneous fibrosis progressed dramatically leading to a pansclerosis, even though the uroporphyrin levels were borderline and the liver enzyme values were normal. Organic solvents are among those substances which can cause a cutaneous fibrosis. The unusually complicated clinical development in our patient was a combination of the two initial factors, the PCT and the long term exposure to organic solvents. The pansclerotic PCT was differentiated from a systemic sclerosis, a disabling pansclerotic morphea and a generalized morphea by means of histological examinations, the absence of a Raynaud phenomenon and the non-involvement of additional organs. Auto-antibodies typical for systemic sclerosis were negative. Using a medium dosage of UVA1 phototherapy and intensive physiotherapy, the progression of the skin disease was stopped and the sclerosis improved. PMID- 12719866 TI - [Benign cellular fibrous histiocytoma with erosion of the phalanx]. AB - Fibrohistiocytic tumors are characterized by the presence of fibroblast like spindle cells and histiocytes. The benign fibrous histiocytoma (dermatofibroma, BFH) as well as the malignant dermatofibrosarcoma protuberans (DFSP) and the malignant fibrous histiocytoma (MFH) belong to this group. A recurrent painful, hard 2 cm tumor on the left hallux of a 54-year-old woman led to an erosion of the underlying phalanx. The patient had suffered from ingrown toenails for more than 10 years. Histologically there was a deep penetrating fibrohistiocytic tumor that grew in a storiform pattern with interspersed foam cells. The tumor was CD34 negative and mitoses were scarce. The diagnosis was benign cellular fibrous histiocytoma (BZFH). BZFH belong to the group of BFH with a high recurrence rate especially after incomplete removal. Damage to the underlying bone has not been reported so far. PMID- 12719867 TI - [Skin and knife. Attributes of St. Bartholomew]. AB - St. Bartholomew, who was born in Kana/Galilee in the first century, is known as one of the twelve apostles of Jesus and is associated with skin diseases due to the attribute "skin" which he carries over his arm like a coat or blanket. Curing the daughter of King Polymios in Armenia led to his doom. He was flayed alive and subsequently crucified. Michelangelo's Last Judgment (1536-1541) in the Sistine Chapel of the Vatican shows him as flayed and holding his own skin in his hand. Today he is regarded as the patron saint of dermatology because of his association with the skin. PMID- 12719868 TI - [Erythematous plaque on the lower eyelid of a 42-year-old patient]. PMID- 12719869 TI - [Reducing trauma with minimally invasive surgery. Evidence and new strategies]. AB - Minimally invasive surgery has become a viable alternative to conventional surgery. The technical advantages of minimally invasive surgery can be translated into clinical benefits for the patients, i.e., less postoperative pain and impairment of lung function, better cosmetic results, shorter hospitalization, and earlier convalescence. Laparoscopic operations have replaced a significant proportion of open surgical procedures and are now routinely used. While the role of laparoscopic surgery has been generally accepted for the management of benign disorders, there is ongoing debate regarding the adequacy of this technique in surgical oncology. There is evidence that minimally invasive surgery can reduce perioperative morbidity in cancer patients. However, definite validation of these procedures for tumor surgery is not yet available due to the lack of prospective randomized trials providing reliable long-term data on disease-free survival and overall survival. It seems likely that minimally invasive procedures will play an important role for the treatment of preneoplastic lesions and tumors of limited size. PMID- 12719871 TI - [New aspects for minimally invasive interventions in orthopedic trauma surgery]. AB - The minimally invasive method has gained complete acceptance for the treatment of diaphyseal fractures. In the majority of cases, this procedure is combined with stabilization of the fracture with insertion of an intramedullary implant. In cases of metaphyseal fracture, use of minimally invasive techniques depends on the extent of simultaneous involvement of the articular surface. For joint fractures, the main goal is anatomic reconstruction. At the same time as these less invasive surgical procedures and markedly improved methods of visualization have evolved, the implants themselves have also advanced. The standard implant of the future for plate osteosynthesis will be the interlocking plate. Minimally invasive surgical techniques will certainly continue to gather ground in the future for the entire field of accident surgery. Reduction of the surgical approach means less postoperative pain for the patient, earlier mobilization, and shortening of hospital stay and the whole process of rehabilitation. PMID- 12719870 TI - [Limitations of laparoscopic visceral surgery in oncology]. AB - Controversial discussion focuses on the application of laparoscopic curative resection for cancer due to oncologic radicality,tumor cell dissemination, and port site metastases. Considering the limitations of laparoscopic surgery, it is necessary to objectively evaluate whether laparoscopic surgery is associated with an improved quality of curative treatment. Therefore, controlled studies comparing the results of laparoscopic vs conventional cancer surgery are mandatory. To date, comparable findings on short-term outcome of laparoscopy with open resection can only be shown for colorectal cancer. However, long-term data including recurrence and survival are still missing as randomized studies (phase IIIb) are still to be completed. Consequently, laparoscopic curative resection should only be performed within controlled trials. In terms of the upper GI tract, minimally invasive surgery has proven to be technically feasible in expert centers including limited resections for early gastric cancer, left pancreatectomies, or hepatic resections for malignancy. Finally, laparoscopy has gained acceptance in the field of diagnosis (e.g., staging laparoscopy, laparoscopic ultrasound) and palliative treatment (e.g. gastroenterostomy, thermoablation) without the need for controlled studies. PMID- 12719872 TI - [Thorascopic coronary surgery. Current status and future perspectives]. AB - Thorascopic coronary surgery has achieved great progress in the past few years. After computer-directed instrument systems were introduced, which enabled high precision and expanded mobility in close quarters, the limitations of conventional endoscopic instruments were largely overcome. Employing modern surgical telemanipulators and endoscopic stabilizers, entirely endoscopic myocardial revascularization of the anterior wall is being successfully performed during off-pump surgery. Longer operating times and conversion rates in about one fifth of the patients due to technical and anatomical limitations still restrict the use of these new surgical techniques compared to conventional surgery. Development of further technologies and integration of video-assisted navigation systems will broaden the application of computer-assisted instrument systems in the near future. PMID- 12719873 TI - [Minimally invasive and intraoperative endoluminal vascular surgery. Critical overlapping and differentiation]. AB - There are only four therapeutic options in minimally invasive vascular surgery. Combined open and endoluminal vascular repair is used most often to minimize surgical trauma. This procedure carries the risk of unfavorable long-term results, which must be hazarded in most elderly and critically ill patients. These new therapeutic procedures require a new generation of vascular surgeons. Besides endovascular surgery there is a new and interesting use of laparoscopic aortic surgery, which competes with endovascular surgery. Also endoscopic vein harvesting for bypass surgery is an interesting procedure to minimize surgical trauma. Minimally invasive procedures for varicose vein disease must be considered fashionable surgery and not aimed at minimizing surgical trauma since cosmetic aspects play an important role. CONCLUSIONS: Minimally invasive surgery does not yet play a major part in vascular surgery. Only combined open and endoluminal vascular repair is on the advance in vascular departments. Because of the increases in the incidence of vascular diseases and the use of new expensive therapeutic procedures that drain health care resources on the one hand and because of the controversy involved in allocating patients to different vascular specialists on the other hand, there is a large amount of dynamite in vascular politics that gives us a thrill for the future! PMID- 12719874 TI - [Endoscopic thoracic surgery: indications, feasibility, and limitations]. AB - Mediastinoscopy and mediastinotomy are used primarily in the staging of lung cancer; they are also of value in biopsying mediastinal masses and lymph nodes to establish diagnoses such as sarcoidosis,lymphoma, and mediastinal tumors. Video assisted thoracic surgery (VATS) was introduced in 1990 and has now replaced open thoracotomy in the evaluation of many pleuropulmonary disorders. Specific advantages of VATS over open thoracotomy include the use of smaller incisions, reduced operative morbidity, and optimal visualization of the entire lung and pleural space. In many centers it has become the procedure of choice for the biopsy of interstitial disease, indeterminate lung nodules, or pleural lesions. The role of VATS for staging of lung cancer patients is still under debate. VATS procedures have also been adopted for the treatment of a wide range of thoracic disorders. With increasing experience surgeons have become more skilled with this limited access technique and meanwhile lobectomies can be performed safely. The role of VATS in the management of lung metastases or lung cancer is still being investigated. It is a concern that there is a temptation to do less when a minimal access operation is performed that does not allow for palpation of the lung. In addition, lymph node dissection cannot be performed adequately and there continue to be reports of local recurrences in port sites. If the VATS approach is to be used, surgeons should always respect the oncological principles that have been developed over the past decades. PMID- 12719875 TI - [Reducing trauma by minimally invasive surgery from the viewpoint of pediatric surgery]. AB - Reduced insufflation pressure during laparoscopy in pediatric surgery can contribute considerably to reducing postoperative pain. Technical innovations in high-grade instruments and thin optics have led to increased diversification of minimally invasive surgery and reduced the strain on patients. The minimally invasive approach is particularly advantageous when the large surgical approaches of conventional procedures can be avoided. The reduction of postoperative adhesions with minimally invasive surgery is a very important factor. The tendency toward decreased infections of surgical wounds is beneficial as well as the improved cosmetic results. The duration of hospital stays and need for analgesics in pediatric surgery have not evidenced any significant differences from conventional procedures. In cases of larger intra-abdominal interventions, it becomes apparent that these small patients can receive oral alimentation at an earlier time. PMID- 12719877 TI - [Surgeons learn how to learn. Study of 76,499 herniorrhaphies performed between 1993 and 1997 registered by the chamber of physicians in Westfalia-Lippe]. AB - INTRODUCTION: New techniques require "new" surgeons who are able to apply them safely. The bushfire-like employment of laparoscopic cholecystectomy confronted a community of surgeons totally inexperienced in that technique. Thus, the collective learning curve was paralleled by a temporary increase of postoperative complications. The aim of the following study was to analyze the learning curve for laparoscopic herniorrhaphy with regard to the complication rates during that period. METHODS: A total of 76,499 questionnaires from the external quality assurance of the Westfalia-Lippe physicians board registered between 1993 and 1997 were analyzed retrospectively. The parameters (duration of the operation and hospitalization, ASA classification, rate of obese patients, intra- and postoperative complications) were analyzed and plotted as a synchronized learning curve of the whole group of surgeons. RESULTS: As for laparoscopic cholecystectomy, a learning curve was found for laparoscopic herniorrhaphy, which could be attributed to a decreasing duration of the procedures and increasing ASA classification and rate of morbid obese patients while complication rates were kept constantly low. After 9 months of application laparoscopic hernia repair seemed to have been introduced to a new collective of surgeons while learning parameters showed inverse trends for a short period of time. The complication rate of laparoscopic hernia repair was constantly lower than in routine open inguinal hernia repair. CONCLUSION: Since laparoscopic inguinal hernia repair is a demanding method, surgeons apparently did not forget the lessons learned from the introduction of laparoscopic cholecystectomy as they were able to avoid increasing intra- and postoperative complication rates during their learning curve. Surgeons are learning to learn. PMID- 12719876 TI - [Importance of rectal extirpation for the therapy concept of low rectal cancers]. AB - INTRODUCTION: The main objective of surgery of rectal carcinomas is to avoid a permanent colostomy by sphincter-sparing surgical procedures. A variety of different abdominoperineal resection rates is described in the literature. MATERIAL/METHOD: The study was performed in 2000 within the framework of a multicentric study including 282 hospitals. The purpose of the study was to document the quality of diagnosis and therapy for colorectal carcinomas.A total of 9477 patients were included in this study: 3402 suffering from a rectal carcinoma and 6075 suffering from a colon carcinoma. RESULTS: A total of 866 abdominoperineal resections was performed. This corresponds to an abdominoperineal resection rate of 27.4%. In 30.4% of all men and in 23.0% of all women an abdominoperineal resection was performed. Of all tumor patients who underwent abdominoperineal resection, 8.3% had a pT4 carcinoma and 57.5% a pT3 carcinoma. Adapted to the localization of the tumor in the rectum, i.e., the distance of the aboral tumor margin to the anal verge, the following abdominoperineal resection rates were found: <4 cm from the anal verge 84.6%, 4 7.9 cm 43.9%, 8-11.9 cm 5.8%, and 12-16 cm 0.5%.Intraoperative complications occurred in 11.8%, specific postoperative complications in 33.1%, and general postoperative complications in 27.4% of the patients. The postoperative lethality was 2.8%. The mean postoperative hospital stay was 21.7 days. Logistic regression identified the body mass index, gender, the distance of the carcinoma from the anal verge, and the T category as independent factors influencing the abdominoperineal resection rate. DISCUSSION: Despite an overall decrease in use, abdominoperineal resection will continue to play an important role for the surgical treatment of low rectal cancers in routine clinical practice in Germany. It will remain an individual decision for each patient whether the tumor and the patient allow sphincter preservation or whether abdominoperineal resection seems to be necessary. According to the results of the present study,a general definition of an abdominoperineal resection rate in an unselected group of patients should be viewed critically. PMID- 12719878 TI - [Validation of the Hannover Score for Polytrauma Outcome (HASPOC) in a sample of 170 polytrauma patients and a comparison with the 12-Item Short-Form Health Survey]. AB - Improved survival rates of patients with multiple injuries have increased general interest in the quality of rehabilitation status after polytrauma. Due to the complex nature of multiple injuries, a special score is needed to evaluate the outcome after polytrauma. The aim of this study was to validate the Hannover Score for Polytrauma Outcome (HASPOC). One hundred seventy patients who had sustained multiple injuries were re-examined using the HASPOC 2 years after trauma. Results were correlated with the 12-Item Short Form Health Survey (SF 12), a validated and established scoring system. The HASPOC was also correlated with clinical parameters predictive of poor outcome. Two control groups consisting of either patients after single injuries or healthy controls were also evaluated using the HASPOC. In the group of patients after multiple injuries, the HASPOC correlated with the SF-12 in injury severity and polytrauma scores, intubation period, hospital stay, and occurrence of injuries below the knee. Differences between healthy controls,patients after single injuries, and patients after multiple injuries were statistically significant when evaluated with the HASPOC. This statistical significance was not achieved using the SF-12. The HASPOC is a valid scoring system and useful for evaluating the rehabilitation status after polytrauma. PMID- 12719881 TI - [Treatment procedures for adenocarcinoma of the esophagogastric junction]. PMID- 12719879 TI - [Synovial sarcoma of the extremities. Results of surgical and multimodal therapy]. AB - BACKGROUND: Synovial sarcoma is a rare tumor frequently occurring in adolescents and young adults. The preferred location, at the distal parts of the extremities, and the high incidence of distant metastases represent major challenges for treatment. The purpose of this study is to analyze the indications for surgery in the context of a multimodal therapy. PATIENTS AND METHODS: Between October 1992 and December 2000, 41 patients were treated surgically for synovial sarcoma of the extremities. Their mean age was 35 years (range: 11-75 years). Extracompartmental tumor growth was present in more than 90% of the patients. Fourteen patients (34%) presented with a tumor size of less than 5 cm (T1) while 27 patients (66%) showed lesions greater than 5 cm (T2). At the time of presentation, seven patients had distant metastases; three located in the lung and four in lymph nodes. RESULTS: Due to a locally unresectable tumor or synchronous distant metastases, 28 patients (68%) underwent preoperative systemic chemotherapy (n=9) or isolated limb perfusion (n=18) or both (n=1). In 29 patients, limb-conserving surgery was possible. Of the 12 patients (29%) who required amputation, 85% had a distal tumor. Sixteen patients received additional postoperative radio- and/or chemotherapy. After a median follow-up of 30 months, only two patients (7%) had developed a local recurrence, while seven patients (20%) suffered from new, distant metastases. The 5-year overall survival rate was 82%. CONCLUSION: With a combined modality treatment, it is possible to achieve excellent local control rates and long-term survival in patients with synovial sarcoma of the extremities. Neoadjuvant treatment can enable limb-sparing surgery in most patients, even if the tumors are located in anatomically difficult areas. PMID- 12719880 TI - [Interdisciplinary cooperation as a prerequisite for precise tumor documentation. Illustrated by an example of surgery for colorectal carcinoma]. AB - Current tumor management is increasingly founded on interdisciplinary cooperation. The main partners in cases of solid tumors are oncologic surgery, medical oncology,and radiotherapy, guided by pathology. The cooperative concept, particularly the individual strategy and selection of the most adequate approach, oriented on guidelines and therapeutic standards, depends on the quality of the involved components as well as personal abilities of the "actors." In addition to the personal qualification, decision making depends on both tumor stage and completeness of tumor removal. In this point, the overall quality of the therapeutic concept is based on an interaction between the operating surgeon and clinical pathologist that had seldom been taken into consideration. The basic rules of their cooperation and quality-focused implementation regarding tumor dignity, stage, and R classification are discussed based on the example of colorectal carcinoma. In particular, those pitfalls in tumor documentation are emphasized,which may appear less relevant for each partner individually, but bear the risk of misinterpretation and therefore misleading conclusions. PMID- 12719882 TI - PF-IND: probability algorithm and software for separation of plant and fungal sequences. AB - The separation of plant and fungal sequences in EST pools by bioinformatic methods is difficult because of sequence similarities between plants and fungi, lack of enough sequence information, and the short length of the isolated fragments. An algorithm and software that utilize the differences in codon usage bias to discriminate between plant and fungal sequences are described. The software (PF-IND) includes five pairs of fungi and their host plants that can be used to analyze a large number of related species. Analysis of a sequence provides an arbitrary value that defines the likelihood that a sequence will be a fungal or a plant gene. The software can distinguish between homologous fungal and plant genes and it helps identify the correct reading frame of unknown expressed sequence tags (ESTs) for which BLAST analyses do not provide clear information. Short sequences of 100-150 bp can be analyzed with high confidence. PF-IND analysis of 100 sequences derived from fungal infected plants identified the origin of 94 sequences. Only 66 sequences were identified by a BLASTX analysis of the same 100 ESTs. Overall, PF-IND is a novel bioinformatic tool aimed at assisting the research of fungus-plant interactions. PMID- 12719884 TI - Occurrence of a pulmonary carcinoid following allogeneic stem cell transplantation for chronic myelogenous leukemia: a case report. AB - Carcinoid tumors are relatively rare neuroendocrine malignancies with an indolent clinical behavior. The majority of cases arise within the gastrointestinal tract, but they may also be encountered in other organs such as the bronchial system. While occurrence of carcinoid tumors has been reported in association with the multiple endocrine neoplasia (MEN) type I syndrome, no clear-cut risk factors have been established for the development of these malignancies. We report the case of a 50-year-old woman who was diagnosed with a pulmonary carcinoid in 2001 after having undergone allogeneic stem cell transplantation for chronic myeloid leukemia (CML) in 1997. This is the first case report of a carcinoid tumor following allogeneic bone marrow transplantation. At the moment, however, an association with CML as well as a causative role of transplantation and intake of immunosuppressants remains speculative. Apart from highlighting the occurrence of a carcinoid in this setting, our case again underscores the importance of nuclear medicine methods, i.e., somatostatin receptor scintigraphy, in staging and follow up of patients with carcinoid tumors. PMID- 12719885 TI - Immune and nonimmune causes of low recovery from leukodepleted platelet transfusions: a prospective study. AB - Alloantibodies against HLA antigens can be reduced by applying leukodepletion to transfusions. Because the importance of immunological and nonimmunological causes of poor platelet transfusion results using leukodepleted transfusions is not clear, we conducted a prospective study in an unselected patient population receiving leukodepleted transfusions. In 97 patients with hematological malignancies, 181 random leukodepleted platelet transfusions were studied for immunological causes of poor platelet transfusion results by calculating the odds ratio of four different screening tests for a low platelet recovery. Nonimmune causes were also studied by calculating the odds ratio of the most prevalent nonimmune causes for a low platelet recovery. No single screening test showed an association with recovery after 1 and 16 h following a platelet transfusion. The combination of a positive enzyme-linked immunosorbent assay (ELISA) and platelet immunofluorescence test (PIFT) or a combination of a positive lymphocyte immunofluorescence test (LIFT) and PIFT, demonstrating an association with a low platelet recovery after 16 h, was present in 2% of all platelet transfusions. Of nonimmune causes, splenomegaly and storage time of platelets for more than 3 days were associated with low platelet recovery after 1 h and 16 h of being present in 29% and 47% of all platelet transfusions, respectively. Immunological causes account for a small proportion of poor platelet transfusion results compared to nonimmunological causes in a nonselected patient population receiving leukodepleted transfusions. PMID- 12719883 TI - Saline, mannitol, and furosemide hydration in acute cisplatin nephrotoxicity: a randomized trial. AB - OBJECTIVE: To determine which hydration (saline, saline + mannitol, or saline + furosemide) is associated with least cisplatin nephrotoxicity. METHODS: We randomized 49 women who received cisplatin (75 mg/m(2) every 3 weeks) into one of the three hydration arms. The 24-h creatinine clearance was measured before and on day 6 after cisplatin infusion. The patients of each arm received 2 l of saline hydration. In the saline + furosemide arm, 40 mg of furosemide was given after hydration. In the saline + mannitol arm, 50 g of mannitol was mixed with the cisplatin. RESULTS: For the first cycle of chemotherapy, 15 women were randomized to saline, 17 to saline + furosemide, and 17 to saline + mannitol. For each group, the creatinine clearances before cisplatin infusion were (means+/-SD, milliliters per minute) 84.5+/-26.8, 82.5+/-24.0 and 87.4+/-25.6, and after cisplatin infusion were 79.1+/-31.9, 68.7+/-21.5, and 56.4+/-22.9, respectively. The decreases in creatinine clearance were similar between the saline group and the saline + furosemide group ( P=0.66), but different between the saline + mannitol group and the saline group ( P=0.02) or the saline + furosemide group ( P=0.02). As each woman received multiple courses of cisplatin, 15 who received saline contributed 41 paired datasets, 17 who received saline + furosemide contributed 49 paired datasets, and 17 who received saline + mannitol contributed 36 paired datasets showed similar patterns. CONCLUSIONS: Hydration with saline or saline + furosemide appears to be associated with less cisplatin nephrotoxicity than saline + mannitol. PMID- 12719887 TI - Successful treatment with reduced-intensity stem cell transplantation in a case of relapsed refractory central nervous system lymphoma. AB - A 33-year-old male with refractory relapsed central nervous system lymphoma underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT) from an HLA-identical sibling after reduced-intensity conditioning chemotherapy. The preparative regimen for allo-HSCT consisted of fludarabine and busulfan. Cyclosporine (CsA) and short-term methotrexate were used as prophylaxis for acute graft-versus-host disease (GVHD). Although CsA was quickly reduced to induce a graft-versus-lymphoma (GVL) effect, no symptoms of GVHD and GVL effect were evident. Donor lymphocyte infusion (DLI) was performed on day +40 following transplantation. The patient developed acute GVHD (grade III) after DLI, and lymphoma regression was observed after the occurrence of GVHD. Four months after transplantation, complete remission was achieved with extensive chronic GVHD, and the patient continues to be disease free at 15 months after transplantation. PMID- 12719886 TI - DNA microarray analysis in malignant lymphomas. AB - Recently, DNA microarray technology has opened new avenues for the understanding of lymphomas. By hybridization of cDNA to arrays containing >10,000 different DNA fragments, this approach allows the simultaneous evaluation of the mRNA expression of thousands of genes in a single experiment. Using sophisticated bioinformatic tools, the huge amount of raw data can be clustered resulting in (1) tumor subclassification, (2) identification of pathogenetically relevant genes, or (3) biological predictors for the clinical course. This approach already has provided novel insights into different entities of B-cell non Hodgkin's lymphomas. Genomic DNA chip hybridization (matrix-CGH) is a complementary approach focussing on genomic aberrations. In this review, we discuss the impact of this new technology both with regard to methodological aspects as well as to novel findings influencing our understanding of lymphomas. PMID- 12719888 TI - Volunteer environmental monitoring and the role of the universities: the case of Citizens' Environment Watch. AB - Universities can provide a stable home for launching collaborative community research projects. Citizens' Environment Watch (CEW), an environmental monitoring initiative based at the University of Toronto, has made significant contributions to environmental education and stewardship in Ontario, Canada. Following dramatic cuts in provincial monitoring programs, citizens and youth have used chemical parameters and biological indicators to gauge water and air quality, and to identify areas requiring remediation and pollution prevention efforts. The relationship of Citizens' Environment Watch to government agencies, funders and other grassroots environmental groups has evolved over the past 5 years as CEW attempts to remain effective without taking on the investigative and enforcement roles to support the regulatory enforcement that has been largely abandoned by government. We explore the challenges inherent in developing and maintaining a volunteer organization that carries out rigorous and useful scientific work and we outline the ability of a university to help overcome these critical challenges. Finally, we present lessons learned for the benefit of other citizen and youth monitoring projects. PMID- 12719889 TI - Knowledge, gender, and resources in small-scale fishing: the case of Honda Bay, Palawan, Philippines. AB - The coastal zone is a place of intense activity where resources, users, and resource-use practices interact. This case study of small-scale fisheries in Honda Bay, Palawan, Philippines shows that resources, space, and gender are intertwined. The study was conducted between June 1997 and July 1998. The data were gathered using free listing, pile sort, ranking, resource mapping, and key informant interviews. The results showed that women's knowledge about fishery resources and their fishing activities are associated with the intertidal zone whereas men's knowledge is associated with coral reefs. In classifying fishery resources, appearance is the main consideration for women whereas a combination of appearance, habitat, and type of fishing gear is the consideration used by men. Market price is very important because of its dependence on the demand of the export market as well as the local market. Women dominate the buying of fishery products. Many women market their husband's catch, process fish, or gather shells and sea cucumber for sale. Among the fishing households, type of fishing gear provides an indication of socioeconomic standing. This paper concludes that access to resources is shaped by gender and age. The differences in resource knowledge possessed by men and women lead to differential access to fishery resources. In addition, the differences in socioeconomic status also influence resource access. The socialization of children into fishing reinforces the gender division of labor and space in the coastal zone. PMID- 12719890 TI - Environmental water-quality zones for streams: a regional classification scheme. AB - Various approaches have been used to classify large geographical areas into smaller regions of similar water quality or extrapolate water-quality data from a few streams to other unmonitored streams. A combination of some of the strengths of existing techniques is used to develop a new approach for these purposes. In this new approach, referred to here as SPARTA (SPAtial Regression-Tree Analysis), environmental characteristics for each monitored stream are first quantified using a Geographic Information System (GIS) and then regression-tree analysis is used to determine which characteristics are most statistically important in describing the distribution of a specific water-quality constituent. GIS coverages of only the most statistically significant environmental characteristics are then used to subdivide the area of interest into relatively homogeneous environmental water-quality zones. Results from the regression-tree analysis not only define the most important environmental characteristics, but also describe how to subdivide the coverage of the specific characteristic (for example, areas with <26% or >or=26% soil clay content). The resulting regionalization scheme is customized for each water-quality constituent based on the environmental characteristics most statistically related to that constituent. SPARTA was used to delineate areas of similar phosphorus, nitrogen, and sediment concentrations (by including land-use characteristics) and areas of similar potential water quality (by excluding land-use characteristics). The SPARTA approach reduced the variability in water-quality concentrations (phosphorus, total nitrogen, Kjeldahl nitrogen, and suspended sediment) within similarly classified zones from that obtained using the US Environmental Protection Agency's nutrient ecoregions. PMID- 12719891 TI - Environmental control and limnological impacts of a large recurrent spring bloom in Lake Washington, USA. AB - A series of statistical analyses were used to identify temporal and spatial patterns in the phytoplankton and nutrient dynamics of Lake Washington, an mesotrophic lake in Washington State (USA). These analyses were based on fortnightly or monthly samples of water temperature, Secchi transparency, ammonium (NH(4)), nitrate (NO(3)), inorganic phosphorus (IP), total nitrogen (TN), total phosphorus (TP), dissolved oxygen (DO), pH and chlorophyll a (chl a) collected during 1995-2000 from 12 stations. Lake Washington has a very consistent and pronounced annual spring diatom bloom which occurs from March to May. During this bloom, epilimnetic chl a concentrations peak on average at 10 microg/L, which is 3 times higher than chl a concentrations typically seen during summer stratified conditions. The spring bloom on average comprised 62% diatoms, 21% chlorophytes and 8% cyanobacteria. During summer stratification, diatoms comprised 26% of the phytoplankton community, chlorophytes 37% and cyanobacteria 25%. Cryptophytes comprised approximately 8% of the community throughout the year. Overall, 6 phytoplankton genera (i.e., Aulacoseira, Fragilaria, Cryptomonas, Asterionella, Stephanodiscus, and Ankistrodesmus) cumulatively accounted for over 50% of the community. These analyses also suggest that the phytoplankton community strongly influences the seasonality of NO(3), IP, DO, pH and water clarity. According to a MANOVA, seasonal fluctuations explained 40% of the total variability for the major parameters, spatial heterogeneity explained 10% of variability, and the seasonal-spatial interaction explained 10% of variability. Distinctive patterns were identified between offshore and inshore sampling stations. The results of our analyses also suggest that spatial variability was substantial, but much smaller than temporal variability. PMID- 12719893 TI - Application of multiattribute decision-making methods for the determination of relative significance factor of impact categories. AB - A relative significance factor (f(i)) of an impact category is the external weight of the impact category. The objective of this study is to propose a systematic and easy-to-use method for the determination of f(i). Multiattribute decision-making (MADM) methods including the analytical hierarchy process (AHP), the rank-order centroid method, and the fuzzy method were evaluated for this purpose. The results and practical aspects of using the three methods are compared. Each method shows the same trend, with minor differences in the value of f(i). Thus, all three methods can be applied to the determination of f(i). The rank order centroid method reduces the number of pairwise comparisons by placing the alternatives in order, although it has inherent weakness over the fuzzy method in expressing the degree of vagueness associated with assigning weights to criteria and alternatives. The rank order centroid method is considered a practical method for the determination of f(i) because it is easier and simpler to use compared to the AHP and the fuzzy method. PMID- 12719892 TI - Analysis of phytoplankton community structure using similarity indices: a new methodology for discriminating among eutrophication levels in coastal marine ecosystems. AB - Nine similarity indices based on phytoplankton community structure were examined for their sensitivity to assess different levels of eutrophication. Two phytoplankton data sets, one from an open coastal system and one from a semi enclosed gulf, associated with different nutrient dynamics and circulation patterns were used for evaluating the indices. The results have shown that similarity indices, measuring interspecific association and resemblance of phytoplankton communities between enriched areas and control sites, were effective for detecting spatial and temporal dissimilarities in coastal marine ecosystems. The structure of the oligotrophic habitat as a potential source of ambiguity for the results was discussed, whereas the validity ranges and the potential applicability of this method were deemed to be dependent on the size of the fraction of the common species among the samples, and the similarity of the classification patterns resulted from this subcategory and those extracted from the overall community data. Furthermore, the study provides a new technique based on the use of the "Box and Whisker Plot" designed to distinguish opportunistic and rare phytoplanktonic species. The similarity indices, applied solely to the dominant species abundance, were more sensitive to resolve eutrophic, mesotrophic and oligotrophic conditions. This procedure can be proposed as an effective methodology for water characterization and can also be used as a qualitative tracer for detecting renewal processes of coastal marine ecosystems. PMID- 12719894 TI - Strategic Environmental Assessment of the Effects of European Union's Regional Development Plans in Donana National Park (Spain). AB - The European Union's Structural Funds are implemented by means of Regional Development Plans (RDP), whose regionally scoped environmental assessment is required. We highlight the deficiencies faced by this approach when subregional areas with high conservation natural values are involved and illustrate it with the case of the RDP of Andalusia region on Donana National Park area (Spain). Commissioned by the World Wildlife Fund, a Strategic Environmental Assessment (SEA) of the Andalusian RDP was carried out focusing on Donana and its area of influence. This is a complex space where some of the most environmentally valuable features in the continent coexist with a surrounding intense and multi sector economic activity, threatening its conservation. In the absence of an established sustainability framework in the Region, a "trickle-down" SEA approach evidenced the need to produce a set of environmental, economic and social guidelines for sustainable management of land, against which the RDP objectives were tested for coherence. An "incremental" SEA approach was also tested, which involved the identification of 79 measures and actions stemming from the RDP provisions and other concurrent planning documents reviewed and the qualitative assessment of their individual and cumulative potential impacts on Donana environments. In the light of the results, a set of complementary mitigating measures was proposed for inclusion in tiered stages of the planning process. Measures to avoid, reduce, remedy and monitor the major types of impact were proposed, including provisions for public participation. SEA emerges as an instrument for non-governmental organizations (NGOs) to carry out independent assessment of public development initiatives. PMID- 12719895 TI - Development and evaluation of a Macroinvertebrate Biotic Integrity Index (MBII) for regionally assessing Mid-Atlantic Highlands Streams. AB - The Macroinvertebrate Biotic Integrity Index (MBII) was developed from data collected at 574 wadeable stream reaches in the Mid-Atlantic Highlands region (MAHR) by the U.S. Environmental Protection Agency's (USEPA) Environmental Monitoring and Assessment Program (EMAP). Over 100 candidate metrics were evaluated for range, precision, responsiveness to various disturbances, relationship to catchment area, and redundancy. Seven metrics were selected, representing taxa richness (Ephemeroptera richness, Plecoptera richness, Trichoptera richness), assemblage composition (percent non-insect individuals, percent 5 dominant taxa), pollution tolerance [Macroinvertebrate Tolerance Index (MTI)], and one functional feeding group (collector-filterer richness). We scored metrics and summed them, then ranked the resulting index through use of independently evaluated reference stream reaches. Although sites were classified into lowland and upland ecoregional groups, we did not need to develop separate scoring criteria for each ecoregional group. We were able to use the same metrics for pool and riffle composite samples, but we had to score them differently. Using the EMAP probability design, we inferred the results, with known confidence bounds, to the 167,797 kilometers of wadeable streams in the Mid-Atlantic Highlands. We classified 17% of the target stream length in the MAHR as good, 57% as fair, and 26% as poor. Pool-dominated reaches were relatively rare in the MAHR, and the usefulness of the MBII was more difficult to assess in these reaches. The process used for developing the MBII is widely applicable and resulted in an index effective in evaluating region-wide conditions and distinguishing good and impaired reaches among both upland and lowland streams dominated by riffle habitat. PMID- 12719896 TI - A performance comparison of metric scoring methods for a multimetric index for Mid-Atlantic highlands streams. AB - When biological metrics are combined into a multimetric index for bioassessment purposes, individual metrics must be scored as unitless numbers to be combined into a single index value. Among different multimetric indices, methods of scoring metrics may vary widely in the type of scaling used and the way in which metric expectations are established. These differences among scoring methods may influence the performance characteristics of the final index that is created by summing individual metric scores. The Macroinvertebrate Biotic Integrity Index (MBII), a multimetric index, was developed previously for first through third order streams in the Mid-Atlantic highlands of the United States. In this study, six metric scoring methods were evaluated for the MBII using measures related to site condition and index variability, including the degree of overlap between impaired and reference distributions, relationships to a stressor gradient, within-sample index variability, temporal variability, and the minimum detectable difference. Measures of index variability were affected to a greater degree than those of index responsiveness by both the type of scaling (discrete or continuous) and the method of setting expectations. A scoring method using continuous scaling and setting metric expectations using the 95th percentile of the entire distribution of sites performed the best overall for the MBII. These results showed that the method of scoring metrics affects the properties of the final index, particularly variability, and should be examined in developing a multimetric index because these properties can affect the number of condition classes (e.g., unimpaired, impaired) an index can distinguish. PMID- 12719897 TI - Therapeutic effect of a single peritumoural dose of IL-2 on transplanted murine breast cancer. AB - Interleukin-2 therapy is not clearly effective against breast cancer both in mouse models and in human patients. However, the study of IL-2 therapy of breast cancer remains important, as 3,700 women died from this malignancy in the Netherlands in 2000. Previously we have shown the therapeutical efficacy of a single peritumoural IL-2 application in different experimental models and in veterinary patients. Here we apply this mode of IL-2 therapy to advanced mouse mammary carcinoma models, i.e., severe metastasised tumours in A/Sn mice and non metastasised carcinomas in BALB/c mice. Mice with advanced transplanted mammary carcinomas were given a single peritumoural treatment with 2.5 x 10(6) IU IL-2 at days 10-14 after i.p. or s.c. inoculation of 10(6) carcinoma cells. Within each experiment it was always possible to distinguish relatively slowly and fast growing tumours which allows the therapeutical effect of IL-2 in tumours with different growth rates to be studied. A new approach to analyse results enabled us to show that survival of mice with transplanted, advanced metastasised breast cancer can be significantly improved after a single local treatment with IL-2. Advanced relatively fast i.p and s.c. growing mammary carcinomas seem to be more sensitive to a single IL-2 treatment than relatively slowly growing tumours. IL-2 was most effective against non-metastasised mouse breast cancer. PMID- 12719898 TI - Endosonographic evaluation of c-kit-positive gastrointestinal stromal tumor. AB - BACKGROUND: Endosonographic features of c-kit-positive gastrointestinal stromal tumors (GISTs) were compared with those of leiomyomas and schwannomas. METHODS: Twenty-four patients with gastric mesenchymal tumors who underwent endoscopic ultrasonography (EUS) and surgical treatment were enrolled. GISTs were defined as c-kit (CD117)-positive tumors, leiomyomas as desmin-positive and c-kit-negative tumors, and schwannomas as S-100-positive and c-kit-negative tumors. Invasion to adjacent organs or more than 20 mitotic counts per 50 high power fields indicated malignancy. RESULTS: There were 19 GISTs, three leiomyomas, and two schwannomas. All five malignant tumors were GISTs. A marginal halo was found in 12 of 19 GISTs and in both of the schwannomas, but not in any of the three leiomyomas. The echogenicities of GISTs were low but higher than that of the normal proper muscle layer, whereas those of leiomyomas and schwannomas were usually low. Lobulation of the tumor surface was documented only in GISTs, particularly in malignant ones. The tumor doubling time of a malignant GIST was 9.3 months, and that of six benign GISTs was 18.7 months (range = 10.7-28.0 months). CONCLUSION: Marginal halo and relatively higher echogenicity on EUS might suggest GIST. Marginal lobulation and a short doubling time may be signs of a malignant GIST. PMID- 12719899 TI - Enteroclysis findings of intestinal Behcet disease: a comparative study with Crohn disease. AB - BACKGROUND: The aim of this study was threefold; to define the enteroclysis (EC) findings of intestinal involvement in Behcet disease (BD), to compare these findings with those seen in Crohn disease (CD), and to determine the relation between the duration of BD and severity of the EC findings. METHODS: From 1997 to 2000, 17 BD and 50 CD cases were examined by EC examination. EC was performed with a 13-F balloon catheter via transnasal entubation. Mucosal and mural changes were evaluated. Statistical analysis was performed with the Mann-Whitney U test to determine the relation between duration of BD and severity of the EC findings. P A, IVS13 -121G>A, A643T, P779P and c.3283T>G) of the IL-12Rbeta2 gene, and the development of type 1 diabetes or atopic asthma as representative Th1- and Th2- dominant diseases, respectively. The association study of each polymorphism of the IL-12Rbeta1 or IL-12Rbeta2 gene and type 1 diabetes or asthma showed that these IL-12R genes did not contribute to the development of type 1 diabetes or asthma in the Japanese population. Further analysis in individuals with susceptibility to intracellular pathogens may elucidate the importance of the IL-12R genes. PMID- 12719943 TI - Skeletal scintigraphic appearance of an auto-transplanted osteoarticular plug: epiphyseal transplant. AB - Nuclear medicine bone scan is an essential diagnostic imaging tool both for the diagnosis and staging of bone tumors and in the follow-up of these patients. It is very important that we be able to discriminate between normal variants, changes related to altered physical stress, and recurrent disease in order to interpret the bone scan meaningfully. We wish to report the appearance of the isotope bone scan, technetium 99m-labeled methylene diphosphonate ((99m)Tc-MDP), associated with an auto-transplanted osteoarticular plug (epiphyseal transplant) performed following limb amputation. This reconstructive surgery can give a potentially misleading appearance on the nuclear medicine bone scan if one is unfamiliar with this surgical technique. PMID- 12719944 TI - Heterotopic gastric mucosa in the gallbladder. PMID- 12719945 TI - Atypical meningioma and extensive calvarium defects in neurofibromatosis type 1. AB - A 9-year-old girl with neurofibromatosis type 1 (NF1) presented with a massive atypical meningioma and calvarial defect. Skull radiographs and cranial CT showed an extensive lytic bone lesion at the vertex. MRI demonstrated a large mass invading the calvarium and sagittal sinus. The histopathological and immunohistochemical diagnosis of the resected mass was atypical meningioma. To our knowledge, this is the first case of NF1 associated with atypical meningioma and massive calvarial defect in a child. PMID- 12719947 TI - The protective effects of physiological and pharmacological concentrations of melatonin on renal ischemia-reperfusion injury in rats. AB - Reactive oxygen species have been implicated in the pathophysiology of renal ischemia reperfusion (I/R) injury. The pineal secretory product melatonin is known to be a potent free radical scavenger and antioxidant. This study was designed to investigate the effects of physiological and pharmacological concentrations of melatonin on I/R injury. Rats were pinealectomized (Px) or sham operated (control) 2 months before the I/R studies. There were eight groups of eight rats each. After a right nephrectomy to produce damage, left renal vessels were occluded for 60 min, followed by 24 h reperfusion, in rats. Malondialdehyde (MDA) levels resulting from I/R were significantly higher in the pinealectomized rats than in the control group. Melatonin administration (4 mg kg(-1) i.p. either before ischemia or reperfusion) to Px and sham-operated rats significantly reduced the MDA values and returned them to the control values. Morphological changes in the groups were similar to the MDA levels. Serum levels of blood urea nitrogen and creatine were unchanged. These results suggest that physiological and pharmacological melatonin concentrations are important for the reduction of I/R-induced damage. We also demonstrated that melatonin, even when administrated just before reperfusion, had a protective effect on I/R injury. It would seem valuable to test melatonin in clinical trials for the prevention of possible I/R injury. PMID- 12719948 TI - Protective role of heparin/heparan sulfate on oxalate-induced changes in cell morphology and intracellular Ca2+. AB - Alterations in intracellular Ca2+ ([Ca2+]i) are generally associated with cellular distress. Oxalate-induced cell injury of the renal epithelium plays an important role in promoting CaOx nephrolithiasis. However, the degree of change in intracellular free calcium ions in renal epithelial cells during oxalate exposure remains unclear. The aim of this study is to determine whether acute short-term exposure to oxalate produces morphological changes in the cells, induces a change in cytosolic Ca2+ levels in renal tubular epithelial cells and whether the application of extracellular glycosaminoglycans (GAGs) prevents these changes. Cultured Mardin-Darby canine kidney cells were exposed to oxalate, and changes in cytosolic Ca2+ were determined under various conditions. The effect of heparin and heparan sulfate (HS) during oxalate exposure was examined. The change in the GAG contents of the culture medium was also determined. Transmission electron microscopy (TEM) was performed for morphological analysis. The degree of change in cytosolic Ca2+ strongly correlated with oxalate concentration. Cytosolic Ca2+ levels decreased in parallel with an increase in the concentration of oxalate. However, this decrease was strongly inhibited by pretreatment with heparin or HS. TEM revealed cytoplasmic vacuolization, the appearance of flocculent material and mitochondrial damage after oxalate exposure. On the other hand, pretreatment with heparin or HS completely blocked these morphological changes. The present data suggest that acute exposure to a high concentration of oxalate challenges the renal cells, diminishes their viability and induces changes in cytosolic Ca2+ levels. Heparin and HS, which are known as potent inhibitors of CaOx crystallization, may also prevent oxalate-induced cell changes by stabilizing the cytosolic Ca2+ level. PMID- 12719949 TI - Is enalapril adequate for the prevention of renal tissue damage caused by unilateral ureteral obstruction and/or hyperoxaluria? AB - Unilateral ureteral obstruction (UUO) and hyperoxaluria (HOX) can lead to end stage renal disease with tubulointerstitial fibrosis. We investigated the effects of enalapril (E), an ACE-inhibitor, on rat kidneys with either UUO or HOX. Sham operated, UUO, HOX, UUO+HOX, UUO+E and HOX+E rats were killed 14 days after UUO and/or HOX was initiated. Rat kidney sections were histologically scored for tissue damage and monocyte/macrophage infiltration was demonstrated with ED1 antibody and measured by computer image analysis software. Serious glomerular and tubulointerstitial damage was found for UUO and HOX, consisting of glomerular basement membrane thickening, tubular dilatation/collapse, tubular basement membrane thickening and the infiltration of mononuclear leucocytes (mainly macrophages). For HOX, calcium oxalate crystals were visible. Neither the scored histological parameters nor monocyte/macrophage infiltration was significantly decreased when E-treated were compared with untreated groups. We conclude that E did not ameliorate the parameters scored in either UUO or HOX. This being contrary to findings by other research groups, we hypothesize that E may be effective only in short-term UUO/HOX, with transforming growth factor, TGF-beta1, formation becoming partly independent of Ang II in late-stage UUO/HOX, or other fibrogenic cytokines than TGF-beta1 becoming predominant. PMID- 12719950 TI - Vascular endothelial growth factor gene polymorphism is associated with calcium oxalate stone disease. AB - Growth factor-related genes regulate cell growth, differentiation and apoptosis in the kidney in response to cellular injury. One of the theories of stone formation is that cellular injury, and thus growth factors, play a role. We therefore investigated the association between growth factor genes and calcium oxalate stone disease. The most frequently seen polymorphism of the vascular endothelial growth factor (VEGF) gene is Bst U I C/T, which is located upstream at the -460th nucleotide. Other growth factor-related gene polymorphisms include the cytochrome P450c17alpha enzyme (CYP17) gene MspA I C/T polymorphism at the 5' UTR promoter region, the epidermal growth factor receptor (EGFR) gene Bsr I polymorphism (A to T) at position 2,073, and the insulin-like growth factor-2 (IGF-2) gene Apa I A/G at exon 9. All four polymorphisms were used as genetic markers in this study in the search for an association between stone disease and growth factor related genes. A normal control group of 230 healthy people, and 230 patients with calcium oxalate stone, were examined. The polymorphism was seen following polymerase chain reaction based restriction analysis. The result revealed a significant difference between normal individuals and stone patients (P=0.0003, Fisher's exact test) in the distribution of the VEGF gene polymorphism as well as an odds ratio of 1.30 (95% confidence interval=0.993-1.715) per copy of the "T" allele. Whereas, the IGF-2, EGFR and CYP17 gene polymorphisms did not reveal a significant association with stone disease. We conclude that the VEGF gene Bst U I polymorphism is a suitable genetic marker of urolithiasis. PMID- 12719951 TI - CT angiography, MR angiography and rotational digital subtraction angiography for volumetric assessment of intracranial aneurysms. An experimental study. AB - The purpose of our experimental study was to assess the accuracy and precision of CT angiography (CTA), MR angiography (MRA) and rotational digital subtraction angiography (DSA) for measuring the volume of an in vitro aneurysm model. A rigid model of the anterior cerebral circulation harbouring an anterior communicating aneurysm was connected to a pulsatile circuit. It was studied using unenhanced 3D time-of-flight MRA, contrast-enhanced CTA and rotational DSA angiography. The source images were then postprocessed on dedicated workstations to calculate the volume of the aneurysm. CTA was more accurate than MRA (P=0.0019). Rotational DSA was more accurate than CTA, although the difference did not reach statistical significance (P=0.1605), and significantly more accurate than MRA (P<0.00001). CTA was more precise than MRA (P=0.12), although this did not reach statistical significance. Rotational DSA can be part of the diagnosis, treatment planning and support endovascular treatment of intracranial aneurysms. The emerging endovascular treatment techniques which consist of using liquid polymers as implants to exclude aneurysms from arterial circulation would certainly benefit from this precise measurement of the volume of aneurysms. PMID- 12719953 TI - Cerebral blood flow, blood volume, and vascular permeability of cerebral glioma assessed with dynamic CT perfusion imaging. AB - We report dynamic CT perfusion imaging assessment of hemodynamics in a patient with a high-grade cerebral glioma and compare our results to those of previously published studies. PMID- 12719952 TI - Diffusion-weighted imaging provides support for secondary neuronal damage from intraparenchymal hematoma. AB - It is controversial whether an intracerebral hematoma (ICH) causes ischemia of surrounding brain. By virtue of its high sensitivity to acute cerebral infarction, diffusion-weighted imaging (DWI) helps answer this question. We used this technique to assess the parenchyma surrounding ICH for restricted diffusion. Echoplanar DWI (b 1000 s/mm(2)) and conventional MRI sequences were performed in 30 subjects (symptom duration 7-75 h) with primary ICH, mean volume: 13+/-15 cm(3). We calculated mean apparent diffusion coefficients (ADC) within high signal regions around the hematoma on DWI or T2-weighted images and within the ICH itself, comparing them to the contralateral brain. We used the Student's t test to examine for differences between these regions and linear regression to relate changes to the age of the ICH. A thin rim of high signal on DWI and a wider rim on T2-weighted images surrounded all hematomas. The ADC within the rim on DWI showed a maximum reduction of 40%, in two patients imaged within 10 h of symptom onset. They rose during the first day (r(2)=0.84; P <0.03) and then showed a mild decrease, becoming the same as ADC in other areas of the brain (r(2)=0.5; P <0.03). The rim on T2-weighting showed a mean increase of 50% and ADC within the ICH were reduced by a mean of 38%; these variations showed no relationship with ICH age and no group showed any relationship with ICH size. The ADC within the three regions was significantly different from each other. The presence of restricted diffusion in the parenchyma surrounding ICH provides support for secondary neuronal damage. PMID- 12719954 TI - Complexation as the most important factor in the fate and transport of heavy metals in the Dnieper water bodies. AB - The results of long-term investigations of the concentrations of dissolved forms of some heavy metals (Mn, Cu, Zn, Pb, Cr, Cd) and their species in the water of the Dnieper reservoirs and the Dnieper-Bug estuary are considered. Chemiluminescent methods, anodic stripping voltammetry, membrane filtration, ion exchange, and gel-permeation chromatography were used for study of the speciation of the metals. It has been found that binding of heavy metals into complexes with dissolved organic matter (DOM) is the dominant factor of their stabilization in solution. The molecular weight distribution of organic metal complexes and their chemical nature, as well as the potential complexing ability of DOM were investigated. Humic substances, particularly fulvic acids, play a major role in the complexation. These ligands bind from 45 to 80% of metals in the form of organic complexes. Metal complex compounds of relatively low molecular weight (<5 kDa) predominated in the organic complexes. PMID- 12719956 TI - Monolithic columns for liquid chromatography. PMID- 12719955 TI - Development and validation of HPLC method for the determination of alpha tocopherol in human erythrocytes for clinical applications. AB - In this work, a simple isocratic reversed-phase HPLC method for determination of alpha-tocopherol in human erythrocytes has been developed and validated. After separation of plasma the erythrocytes were washed three times with 0.9% sodium chloride containing 0.01% butylated hydroxytoluene (BHT) as antioxidant and then were diluted 1:1 (v/v) with the same solution. In the liquid-liquid extraction (LLE) procedure, 2500 microL of n-hexane was added to 500 microL of erythrocytes. After 2 min this mixture was deproteinized by addition of cool ethanol (500 microL, 5 min) denatured with 5% methanol containing alpha-tocopherol acetate (20 micromol L(-1)), as internal standard, and then extracted for 5 min by vortex mixing. After centrifugation (10 min, 1600xg) an aliquot (2000 microL) of the clean extract was separated and evaporated under nitrogen. The residue was dissolved in 400 microL methanol and analysed by reversed-phase HPLC on a 4.6 mmx150 mm, 5 microm Pecosphere C18 column; the mobile phase was 100% methanol, flow rate 1.2 mL min(-1). The volume injected was 100 microL and detection was by diode-array detector at a wavelength of 295 nm. The extraction recovery of alpha tocopherol from human erythrocytes was 100.0+/-2.0%. The detection limit was 0.1 micromol L(-1) and a linear calibration plot was obtained in the concentration range 0.5-20.0 micromol L(-1). Within determination precision was 5.2% RSD (n=10), between determination precision was 6.1% RSD (n=10). The method was applied successfully in a clinical study of patients with acute pancreatitis and for determination of the reference values in the healthy Czech population. PMID- 12719957 TI - Analysis by partial reflection spectrometry of protonated tetraphenylporphyrin adsorbed at a liquid-liquid interface. AB - Visible reflection spectra of diprotonated meso-tetraphenylporphyrin adsorbates spontaneously formed at a dodecane-aqueous sulfuric acid interface have been measured using a home-made device comprising a prism-cell and variable-angle optics. The tilt angle of the pyrrole ring plane was estimated to be 47 degrees from the interface normal by use of an experimentally evaluated molecular density (1.20x10(-10) mol cm(-2)) of the diprotonated molecule in a monolayer form at the liquid-liquid interface. Positive and negative bands have been observed in the p polarized partial internal reflection (p-PIR) spectra, whose band locations correspond to those in p-polarized external reflection (p-ER) spectra. Nevertheless, the bands in the p-PIR exhibited reversed sign to those of p-ER spectra. These suggest that the surface selection rule of the p-PIR spectrometry has a reversal rule of p-ER and p-PIR can also be used for the analysis of molecular orientation. PMID- 12719958 TI - Capillary electrophoretic separation of dsDNA under nonuniform electric fields. AB - Improved sensitivity for the analysis of DNA by capillary electrophoresis has been achieved, based on simultaneous increases in optical path length and injection volume. To increase the optical path length, bubble cells with diameters ranging from 150 to 450 microm have been fabricated and tested. In terms of resolution and sensitivity, a bubble cell of 300 microm diameter is appropriate when using 75-microm capillaries. To allow greater injection volumes, we performed on-line concentration of DNA in the presence of electroosmotic flow (EOF) using 2.0% poly(ethylene oxide) (PEO). With a 300-microm bubble cell, a 170 fold improvement in the sensitivity for the 89-bp fragment has been accomplished when injecting about 0.33 microL DNA. In the presence of the bubble cell, the resolution for the large fragments improves while that for the small ones (<124 base pair) decreases. The effect of bubble cells was further investigated by conducting DNA separation in the absence of EOF, showing that improvements in resolution are mainly due to increased migration differences when DNA migrated at low electric field strengths in the bubble region. We have suggested that such an effect is more profound using shorter capillaries, leading to complete separation of phiX 174 RF DNA-Hae III digest in 2 min. PMID- 12719959 TI - Process analytical chemistry-future trends in industry. PMID- 12719961 TI - Discriminative stimuli that control instrumental tobacco-seeking by human smokers also command selective attention. AB - RATIONALE: Incentive salience theory states that acquired bias in selective attention for stimuli associated with tobacco-smoke reinforcement controls the selective performance of tobacco-seeking and tobacco-taking behaviour. OBJECTIVES: To support this theory, we assessed whether a stimulus that had acquired control of a tobacco-seeking response in a discrimination procedure would command the focus of visual attention in a subsequent test phase. METHODS: Smokers received discrimination training in which an instrumental key-press response was followed by tobacco-smoke reinforcement when one visual discriminative stimulus (S+) was present, but not when another stimulus (S-) was present. The skin conductance response to the S+ and S- assessed whether Pavlovian conditioning to the S+ had taken place. In a subsequent test phase, the S+ and S- were presented in the dot-probe task and the allocation of the focus of visual attention to these stimuli was measured. RESULTS: Participants learned to perform the instrumental tobacco-seeking response selectively in the presence of the S+ relative to the S-, and showed a greater skin conductance response to the S+ than the S-. In the subsequent test phase, participants allocated the focus of visual attention to the S+ in preference to the S-. Correlation analysis revealed that the visual attentional bias for the S+ was positively associated with the number of times the S+ had been paired with tobacco-smoke in training, the skin conductance response to the S+ and with subjective craving to smoke. Furthermore, increased exposure to tobacco-smoke in the natural environment was associated with reduced discrimination learning. CONCLUSIONS: These data demonstrate that discriminative stimuli that signal that tobacco-smoke reinforcement is available acquire the capacity to command selective attentional and elicit instrumental tobacco-seeking behaviour. PMID- 12719960 TI - Escitalopram, the S-(+)-enantiomer of citalopram, is a selective serotonin reuptake inhibitor with potent effects in animal models predictive of antidepressant and anxiolytic activities. AB - OBJECTIVE: The pharmacological profile of escitalopram, the S-(+)-enantiomer of citalopram, was studied and compared with citalopram and the R-(-)-enantiomer, R citalopram. METHODS: Inhibition of the serotonin transporter (5-HTT) was studied in COS-1 cells expressing the human 5-HTT (h-5-HTT) and in rat brain synaptosomes. In vitro selectivity was studied relative to noradrenaline transporter (NAT) and dopamine transporter (DAT) function in rat brain synaptosomes, and affinities for other binding sites were determined. In vivo 5 HT activity was measured as inhibition of neuronal firing rate in rat dorsal raphe nucleus (DRN) and enhancement of 5-hydroxytryptophan (5-HTP)-induced behaviour (mouse and rat). Furthermore, studies were conducted in models of antidepressant (mouse forced-swim test), anxiolytic [foot-shock-induced ultrasonic vocalization (USV) in adult rats and mouse black and white box] and anti-aggressive activity (socially isolated mice). RESULTS: Escitalopram inhibited 5-HTT functions approximately 2 times more potently than citalopram and at least 40 times more potently than R-citalopram. Escitalopram showed insignificant activity at other monoamine transporters and 144 other binding sites. Escitalopram inhibited 5-HT neuronal firing in DRN and potentiated 5-HTP induced behaviours more potently than citalopram; R-citalopram was inactive. Escitalopram and citalopram, but not R-citalopram, reduced forced-swimming induced immobility and facilitated exploratory behaviour in the black and white box. Escitalopram and citalopram inhibited USV potently; R-citalopram was several times less potent. Escitalopram, citalopram and R-citalopram inhibited aggressive behaviour weakly. Escitalopram and citalopram had very potent anti-aggressive effects when co-administered with l-5-HTP. CONCLUSION: Escitalopram is a very selective 5-HT reuptake inhibitor. It is more potent than its racemate citalopram and is effective in animal models predictive of antidepressant and anxiolytic activities. PMID- 12719963 TI - Acute administration of nutritionally sourced tryptophan increases fear recognition. AB - RATIONALE: The serotonin precursor tryptophan (TRP) has been widely used as a nutritional supplement and antidepressant. Recently, however, the use of TRP has been severely restricted due to its association with the eosinophilic myalgic syndrome, an autoimmune disorder probably caused by ingestion of a contaminant produced in certain TRP manufacturing processes. OBJECTIVES: To determine the bioavailability of a nutritional source of TRP obtained from milk protein and to assess whether administration of this material produced neuroendocrine and neuropsychological effects consistent with increased brain serotonin activity. METHODS: We studied 24 healthy subjects who ingested approximately 1.8 g of nutritionally-sourced TRP or placebo in a double-blind, parallel group, design. We carried out venous sampling for amino acid and hormone estimation and performed a test of emotional processing using a facial expression recognition task. RESULTS: The nutritionally-sourced TRP caused a substantial increase in the availability of TRP in plasma. Relative to placebo the TRP material produced some evidence of an increase in plasma cortisol, and enhanced the perception of fearful and happy facial expressions. CONCLUSIONS: A nutritional source of TRP increased the availability of TRP for brain serotonin synthesis and produced endocrine and neuropsychological changes consistent with increased brain serotonin function. The effect of TRP on emotional processing may be relevant to its reported activity in primate studies of social behaviour. PMID- 12719962 TI - Comparison of reinstatement of ethanol- and sucrose-seeking by conditioned stimuli and priming injections of allopregnanolone after extinction in rats. AB - RATIONALE AND OBJECTIVES: Understanding the mechanism of relapse provoked by conditioned and unconditioned stimuli is critical to improving treatments for alcoholism. This study compared the reinstatement of alcohol- or sucrose-seeking by conditioned stimuli and priming injections of the neuroactive steroid, allopregnanolone (ALLO). METHODS: Rats were trained to lever-press for 0.1 ml of 10% ethanol or 5% sucrose solutions. Responding was then extinguished, and subjects were tested for reinstatement of lever-press responding. The effects of priming injections of 0, 1.0, 3.0 and 7.5 mg/kg ALLO were determined in subjects trained to self-administer ethanol, and the response-reinstating effects of priming injections of 3.0 mg/kg ALLO were compared with those of conditioned cue presentation in subjects trained to self-administer either ethanol or sucrose. RESULTS: Priming injections of ALLO dose-dependently reinstated previously extinguished responding for ethanol, as shown by increased responding on the active (ethanol) lever. Contingent presentation of cues previously associated with the reinforcer increased the number of active lever-presses for both ethanol and sucrose- trained subjects. In contrast, pretreatment with 3.0 mg/kg ALLO increased the number of active lever-presses for subjects that were trained to self-administer ethanol, but not sucrose. CONCLUSIONS: ALLO promotes responding for ethanol, but not sucrose, following a period of abstinence, suggesting that GABA(A) receptor modulation may contribute to processes involved in reinstatement of ethanol-seeking behavior. In contrast, conditioned stimuli reinstate previously extinguished ethanol- and sucrose-seeking behavior, indicating that the mechanisms that subserve cue-induced reinstatement do not depend upon the nature of the positive reinforcer. PMID- 12719964 TI - Hypericum perforatum attenuates nicotine withdrawal signs in mice. AB - RATIONALE: Hypericum perforatum is used as a natural antidepressant, and other antidepressants have been marketed to aid in smoking cessation. OBJECTIVE: We investigated the effects of an extract of Hypericum perforatum (Ph-50) on withdrawal signs produced by nicotine abstinence in mice. METHODS: Nicotine (2 mg/kg, four injections daily) was administered for 14 days to mice. Different doses of Ph-50 (125-500 mg/kg) were administered orally immediately after the last nicotine injection. In another experiment, Ph-50 (500 mg/kg) was orally administered in combination with nicotine, i) starting from day 8 until the end of the nicotine treatment period, or ii) during nicotine treatment and after nicotine withdrawal, or iii) immediately after the last nicotine injection. On withdrawal from nicotine, all animals were evaluated for locomotor activity and abstinence signs. RESULTS: The locomotor activity reduction induced by nicotine withdrawal was abolished by Ph-50, which also significantly and dose-dependently reduced the total nicotine abstinence score when injected after nicotine withdrawal. CONCLUSIONS: These data show that treatment with Hypericum perforatum attenuates nicotine withdrawal signs in mice. Further studies are necessary to test the possibility that it may be used for smoking cessation treatment in humans. PMID- 12719966 TI - Guidelines for informed consent in biomedical research involving paediatric populations as research participants. AB - To promote and protect the best interests of children involved in biomedical research, paediatricians have to ensure that participating minors and their parents/legal representatives have understood and assented/consented to the research. Therefore guidelines providing child-specific guidance that are compatible with other international guidelines on informed consent are laid down. Special regard is paid to the willingness to participate and the social and cultural background of the patients, the legal conditions of the countries, the capacity of the child to understand and give his/her informed assent, the adequate communication with the child and the parents, the respect of the will of the patient, the understandable written informed consent of legal representatives and to the evaluation of the informed consent/assent process by competent ethics committees. PMID- 12719965 TI - Needed: mouse/human cross validation of reinstatement/relapse models (and drug reward models) to model human substance abuse vulnerability allelic variants. PMID- 12719967 TI - Stool interleukin 1beta and interleukin 1 receptor antagonist concentrations in children with active ulcerative colitis and during recovery. PMID- 12719969 TI - Radiographic assessment of the gap between oesophageal pouches in infants with oesophageal atresia without fistula. PMID- 12719968 TI - Ligneous conjunctivitis, hydrocephalus, hydrocele, and pulmonary involvement in a child with homozygous type I plasminogen deficiency. AB - Ligneous conjunctivitis is a rare and unusual form of chronic pseudomembranous conjunctivitis which usually starts in early infancy. Plasminogen deficiency has recently been associated with ligneous conjunctivitis. The disease may be associated with pseudomembranous lesions of other mucous membranes in the mouth, nasopharynx, trachea, and female genital tract and also with congenital hydrocephalus. In this report, a 1-month-old Turkish boy who had pseudomembranous conjunctivitis, occlusive hydrocephalus, and hydrocele is presented. After surgery for ventriculo-peritoneal shunt establishment, he developed inspiratory stridor, respiratory distress, and pulmonary atelectasis. Tracheal pseudomembranes were also demonstrated by bronchoscopy. Plasminogen antigen level and plasminogen activity were very low. Genomic DNA from the patient was screened for mutations in the plasminogen gene and a homozygous L650fsX652 mutation (deletion of 2081C) was detected. Both of his parents were heterozygous for this mutation. He died due to respiratory failure during follow-up. CONCLUSION: Ligneous conjunctivitis related to type I plasminogen deficiency is relatively common in the Turkish population, however, mutations are heterogeneous and a common founder is unlikely. PMID- 12719970 TI - The presentation of asymptomatic palpable movable mass in female inguinal hernia. AB - The surgical approach to the inguinal canal in girls is identical to that in boys. The sliding hernia which contains the ovary with or without the fallopian tube occurs occasionally in female patients. In our clinical experience, we found that a hydrocele in the labium, also presenting with an asymptomatic palpable movable mass, mimics a sliding hernia with ovary. In an attempt to differentiate between hydrocele and sliding inguinal hernia with ovary in female patients, we report our experience dealing with the two situations at a single institution within a 5-year period. Between July 1997 and June 2002, 1800 infants and children underwent surgery for inguinal hernia at Chang Gung Children's Hospital, of whom 580 were female infants and girls aged 1 month to 14 years (mean, 5.7 years). Some 32 patients (5.3%) presented with an asymptomatic palpable movable mass over the labium major. Pre-operative sonography was performed for all cases. Twenty-six female infants aged 1 month to 18 months (mean 5 months) had sliding hernia; both the ovary and fallopian tube were contained. Six girls aged 2 years to 6 years (mean 4.6 years), had hydrocele of the canal of Nuck. The accuracy of pre-operative diagnosis with sonography was 100%. CONCLUSION: Sonography is an easy and accurate pre-operative diagnostic procedure. We suggest that sonography be performed routinely in all female cases with an inguinal hernia containing a palpable movable mass. PMID- 12719971 TI - Rapidly developing Cushing syndrome in a 4-year-old patient during combined treatment with itraconazole and inhaled budesonide. AB - A 4-year-old boy with cystic fibrosis developed hypertension, rapid weight gain and a moon face 2 weeks after starting a combined treatment of oral itraconazole and inhaled budesonide for a suspected allergic bronchopulmonary aspergillosis. Adrenal suppression was documented and found to persist 3 months after stopping this combined treatment. CONCLUSION: To the best of our knowledge, this is the first time that an iatrogenic Cushing syndrome in a young child with cystic fibrosis after such combined treatment is reported. The inhibition of cytochrome P4503A by intraconazole and a higher glucocorticoid tissue sensitivity is suggested as the underlying mechanism. PMID- 12719973 TI - Glipizide treatment with short-term alcohol abuse resulting in subfulminant hepatic failure. PMID- 12719972 TI - Malignant conversion of florid oral and labial papillomatosis during topical immunotherapy with imiquimod. AB - We report a case of a 61-year-old woman suffering from florid oral papillomatosis with a squamous-cell cancer of the floor of the mouth, which was removed by scalpel surgery combined with a radical neck dissection in 1996. Between 1996 and 2000 several histologically benign papillomatous lesions of mouth and lips were removed with laser and electrosurgery. However, the lesions recurred. In July 2000 hyperkeratotic, wart-like lesions were present at the lower and upper lips and at the right angle of the mouth and the adjacent oral mucosa. Overnight treatment with a topical 5% imiquimod cream on a Monday-Wednesday-Friday schedule was initiated. However, due to severe irritation and pain the application had to be reduced to 4 h per night, three times a week, followed by a therapy-free interval of 2 weeks. Despite this treatment consisting of four cycles of 3 weeks (1 week treatment and 2 weeks pause), the lesions increased markedly in size. A biopsy taken from the tumorous lesion from the right angle of the mouth proved to be a squamous-cell carcinoma. The tumors of the labial and oral mucosal sites as well as the right submandibular lymph nodes were removed by wide scalpel excision. The lips were reconstructed by plastic surgery. 24 months after surgical intervention no recurrence nor metastasis to lymph nodes or distal sites were observed. PMID- 12719974 TI - Epithelial-myoepithelial carcinoma of the parotid gland-evidence of contrasting DNA patterns in two different histological parts. AB - Epithelial-myoepithelial carcinoma (EMC) is a rare neoplasm arising predominantly in the salivary glands, in particular in the parotid gland. We report the morphological features of an epithelial-myoepithelial carcinoma of the parotid gland with one lymph-node metastasis including a molecular genetic study of this tumor. Immunohistochemical and ultrastructural results confirmed the epithelial myoepithelial dualism of the carcinoma. The loss of heterozygosity (LOH) analysis revealed different LOH results for the solid and the tubular growth pattern of the primary tumor, but showed identical findings for the solid primary tumor component and the lymph node metastasis which had also a solid appearance. LOH could be demonstrated in the whole primary tumor at D13S217 (13q12) and D18S58 (18q21). In three other microsatellite loci [D9S162 (9p22-p21), D10S251 and D10S541 (surrounding the PTEN/MMAC1 gene on 10q23-q24)], clearly recognizable LOH was found in the solid part and in the metastasis, whereas the tubular component demonstrated only a slight decrease of the same allele. No mutation or methylation of the p16 gene or alteration of the PTEN/MMAC1 gene could be found. Nevertheless, our results provoke a discussion, whether these genetic alterations could be considered as determinants of histologically and prognostically divergent types in EMC. PMID- 12719975 TI - Correlation of emmprin expression in vascular endothelial cells with blood-brain barrier function: a study using magnetic resonance imaging enhanced by Gd-DTPA and immunohistochemistry in brain tumors. AB - In a previous study, we demonstrated that the expression levels in tumor cells of emmprin (CD147) correlated with the grade of astrocytic tumors. Also, we found that emmprin was expressed in vascular endothelial cells of the non-neoplastic brain and hypothesized that emmprin expression could be associated with normal blood-brain-barrier (BBB) function of vascular endothelial cells. In this study, this possibility was examined in non-neoplastic brain, glioma and metastatic carcinoma tissues by comparing emmprin immunohistochemistry with gadolinium diethylenetriaminepentaacetic acid (Gd-DTPA) enhancement of magnetic resonance imaging (MRI), which is a clinical indicator of the BBB function. This study included 10 cases of non-neoplastic brain tissues, 7 of metastatic carcinoma, 7 of diffuse astrocytoma, 4 of anaplastic astrocytoma and 13 of glioblastoma multiforme. In all the cases, MRI with administration of Gd-DTPA was performed. The lesions were resected using the microdissection method with the help of ultrasonography and a neuronavigator. The tissues from Gd-DTPA-enhanced or non enhanced areas were processed into frozen sections and subjected to immunohistochemistry with anti-emmprin antibody. The expression of emmprin in brain vascular endothelial cells inversely correlated with Gd-DTPA-enhancement of MRI: emmprin was positive in tissues not enhanced by Gd-DTPA and was negative in DTPA-enhanced tissues. Since BBB function presumably remains unimpaired in regions in which MR images are not Gd-DTPA-enhanced, emmprin expression appears to be associated with unimpaired BBB function. This is the first report to demonstrate a possible correlation between emmprin expression and BBB function in humans. PMID- 12719977 TI - Expression of thrombospondin-1 is correlated with microvessel density in gastric carcinoma. AB - Thrombospondin-1 (TSP-1) has been shown to play a role in angiogenesis in a variety of cancers, but some studies indicated a difference in the mechanism of TSP-1 on neovascularization according to organ or histological type. Wild-type p53 protein has been shown to induce TSP-1 expression. We examined the expression of TSP-1 protein in 80 gastric carcinomas using immunohistochemistry and studied the relationship with microvessel counts, p53 expression and clinicopathological factors. We also performed reverse-transcriptase polymerase chain reaction analysis for the TSP-1 mRNA expression in gastric carcinoma cell lines and gastric cancer tissue after laser capture microdissection. Strong expression of TSP-1 protein was detected in 30 (38%) of the 80 cases. Positive staining for TSP 1 was seen in the cytoplasm of the cancer cells. TSP-1 mRNA expression was confirmed in a majority of gastric carcinoma cell lines and carcinoma tissues. Microvessel counts were significantly higher in tumors with strong TSP-1 protein expression than in those without expression or weak expression of TSP-1 ( P=0.011). No significant correlation was found between TSP-1 expression and p53 staining and clinicopathological factors. Our results support an idea that increased TSP-1 expression may be associated with an angiogenic phenotype in gastric carcinoma and suggest that TSP-1 may play diverse roles in each organ. PMID- 12719976 TI - Vinculin: a novel marker for quiescent and activated hepatic stellate cells in human and rat livers. AB - In liver injuries, the quiescent hepatic stellate cells (HSCs) promptly change to activated HSCs, which are easily identified by the intense immunoreactivity for alpha-smooth muscle actin. However, reproducible markers for quiescent HSCs in formalin-fixed, paraffin-embedded liver tissue sections have not yet been reported. We immunohistochemically examined the expression of vinculin, one major protein of dense plaques, on cultured LI90 cells and on HSCs in normal and diseased human and rat livers. In cultured LI90 cells, vinculin appeared as small linear patches. Although vinculin was consistently negative in the routine liver tissue sections, an antigen retrieval technique using microwave oven heating yielded excellent effects. Using this technique, the formalin-fixed, paraffin embedded human and rat normal liver tissue sections showed the vinculin immunoreactivity along the sinusoidal wall. Immunoelectron microscopic observation of hepatic parenchyma demonstrated that the vinculin was exclusively expressed in quiescent HSCs. In fetal rat livers, vinculin-positive quiescent HSCs gradually increased in number with gestation. In diseased livers the activated HSCs showed more intense immunoreaction for vinculin. These results indicate that, using microwave pretreatment, vinculin is expressed in quiescent and activated HSCs in routinely processed liver tissue sections. It could allow us to evaluate the development and distribution of quiescent HSCs and to examine the relationship between quiescent and activated HSCs. PMID- 12719978 TI - Sepsis-related microvascular myocardial damage with giant cell inflammation and calcification. AB - We report a case of a 27-year-old man who died suddenly 2 days after being discharged from the intensive care unit where he had resided for 25 days due to severe sepsis. On postmortem examination, the findings were restricted to the heart and characterized by microvascular myocardial damage with giant cell inflammation and calcification. The microcirculation seems to be involved in the mechanism of cell injury, very likely due to flow disturbances caused by septic shock. The dystrophic calcification occurred in myocytes showing myocytolysis. Lysozyme produced by macrophages and myocytes in areas of myocytolysis and giant cells appear to function cooperatively and/or synergistically to influence mineralization. In addition, the expression of inducible nitric oxide synthase by myocytes implies that nitric oxide could contribute to myocardial cell damage. Immunolabeling studies of the giant cells confirmed that they are derived from macrophages. PMID- 12719979 TI - Force/velocity curves of fast oxidative and fast glycolytic parts of rat medial gastrocnemius muscle vary for concentric but not eccentric activity. AB - The purpose of this study was to compare the force exerted by the rat medial gastrocnemius (GM) muscle with either fast oxidative or fast glycolytic parts active during concentric and eccentric contractions at different velocities. The proximal end of the GM contains mainly fast oxidative fibres and the distal end predominantly fast glycolytic fibres. Different parts of GM were activated by selective stimulation of nerve branches. Fast oxidative or fast glycolytic muscle parts of anaesthetised male Wistar rats were activated maximally. After assessment of concentric force/velocity (F/v) relations (n=11), some of the muscles were subjected to a fatiguing series of isometric contractions (n=5). Fast oxidative muscle parts showed a significantly lower mean (+/-SD) maximal power output (P(max) 0.12+/-0.06 W) and fatigability than fast glycolytic muscle parts (P(max) 0.20+/-0.06 W). The remaining muscles performed eccentric contractions. The eccentric F/v curves were not significantly different for fast oxidative and fast glycolytic muscle parts (n=6). Maximum eccentric force relative to the maximum isometric force (157+/-3% and 153+/-6% respectively,P=0.99) was reached at a velocity of 60 mm s(-1). It is concluded that eccentric F/v relations of rat GM with either fast oxidative or fast glycolytic parts active are very similar despite the differences in the concentric F/v relations. PMID- 12719980 TI - Inhibition of fast sodium current in rabbit ventricular myocytes by protein tyrosine kinase inhibitors. AB - The present study investigated the effects of protein tyrosine kinase inhibitors on the fast sodium current ( I(Na)) in rabbit ventricular myocytes. Single rabbit ventricular myocytes were isolated enzymatically using Langendorff perfusion. I(Na) was recorded using the whole-cell patch-clamp technique at room temperature. The protein tyrosine kinase inhibitors genistein, AG957, ST638, and PP2 reversibly inhibited I(Na) in a concentration-dependent manner. At a test pulse potential of -30 mV, genistein (n=7) inhibited I(Na) by 37.7+/-3.2%, 53.4+/ 2.5%, and 71.8+/-2.7% at concentrations of 15, 50, and 100 microM, respectively, without changing the voltage dependence of activation, while 100 microM AG957, 100 microM ST638, and 30 microM PP2 inhibited I(Na) by 38.7+/-2.4, 35.8+/-3.4, and 21.1+/-3.9%, respectively. Genistein (100 microM) and AG957 (100 microM) shifted the voltage for half-maximal inactivation of I(Na) from -76.7+/-2.0 mV (n=10) in control to -88.37+/-2.6 mV (n=6, P<0.05), and -82.9+/-1.7 (n=4, P<0.05), respectively, without changing the slope factor. Genistein and AG957 also significantly prolonged the time course of I(Na) recovery from inactivation. Daidzein and PP3, inactive analogs of genistein and PP2, respectively, did not inhibit I(Na) significantly. We conclude that protein tyrosine kinase signaling pathways may play an important role in regulation of I(Na) in cardiac myocytes. PMID- 12719981 TI - Synaptic uptake and beyond: the sodium- and chloride-dependent neurotransmitter transporter family SLC6. AB - The SLC6 family is a diverse set of transporters that mediate solute translocation across cell plasma membranes by coupling solute transport to the cotransport of sodium and chloride down their electrochemical gradients. These transporters probably have 12 transmembrane domains, with cytoplasmic N- and C terminal tails, and at least some may function as homo-oligomers. Family members include the transporters for the inhibitory neurotransmitters GABA and glycine, the aminergic transmitters norepinephrine, serotonin, and dopamine, the osmolytes betaine and taurine, the amino acid proline, and the metabolic compound creatine. In addition, this family includes a system B(0+) cationic and neutral amino acid transporter, and two transporters for which the solutes are unknown. In general, SLC6 transporters act to regulate the level of extracellular solute concentrations. In the central and the peripheral nervous system, these transporters can regulate signaling among neurons, are the sites of action of various drugs of abuse, and naturally occurring mutations in several of these proteins are associated with a variety of neurological disorders. For example, transgenic animals lacking specific aminergic transporters show profoundly disturbed behavioral phenotypes and probably represent excellent systems for investigating psychiatric disease. SLC6 transporters are also found in many non neural tissues, including kidney, intestine, and testis, consistent with their diverse physiological roles. Transporters in this family represent attractive therapeutic targets because they are subject to multiple forms of regulation by many different signaling cascades, and because a number of pharmacological agents have been identified that act specifically on these proteins. PMID- 12719982 TI - Skin care management: educational aspects. AB - Health education is effective in primary, secondary and tertiary prevention of occupational skin disorders, as clearly demonstrated by interventional studies carried out at the University of Osnabrueck. Health education may intervene with (a) habituated behaviour of affected employees and (b) the condition of working environments. In conclusion, prevention of occupational skin diseases and maintenance of health can be achieved by the conditioning of individual behaviour via specific educational programmes in addition to dermatological care. PMID- 12719983 TI - No significant effect of iron deficiency on cadmium body burden or kidney dysfunction among women in the general population in Japan. AB - OBJECTIVE: To examine if iron-deficient conditions modify body burden or health effects of cadmium among women in the general population in Japan. METHODS: In 2002, 1,482 women aged 20 to 74 years in six prefectures in Japan provided informed consent to participate in this study. They offered peripheral blood and spot urine samples, and answered questionnaires on their social habits and health conditions. Never-smoking, non-pregnant and non-lactating healthy women (1,190 subjects) were selected from the volunteers. Blood samples were analyzed for serum iron, ferritin and total iron-binding capacity (TIBC) in addition to red blood cell (RBC) counts and hemoglobin (Hb) concentration as markers of anemia and iron deficiency. Urine samples were analyzed for cadmium (Cd), alpha(1) microglobulin (alpha(1)-MG), beta(2)-microglobulin (beta(2)-MG) as markers of Cd burden and Cd-induced tubular dysfunction; the measures were expressed after being corrected for creatinine (cr) as, e.g., Cd-Ucr. RESULTS: The subjects were classified into anemic (37 women) and iron-deficient (388 women) groups separately from healthy controls (765 women), taking ferritin (<20 ng/ml) and Hb (<10 g/100 ml) as classification indicators. Strictly matched pairs (with regard to age and prefecture) were established for 36 anemic and 280 iron-deficient cases. Comparison between the cases and the matched controls showed that serum iron was lower and TIBC was higher in accordance with lower levels of ferritin and Hb in the anemic and iron-deficient groups, although the RBC count was only slightly reduced (the anemic group) or stayed essentially unchanged (the iron deficient group). In contrast, no significant increase in Cd-Ucr, alpha(1)-MG, or beta(2)-MG was observed in either the anemic group or the iron-deficient group compared with the matched controls. Cd-Ucr in one case of clinical anemia, however, tended to be higher than the levels among women of the same age range and from the same prefecture. Her alpha(1)-MG-Ucr and beta(2)-MG-Ucr, however, remained un-elevated. CONCLUSIONS: The current level of iron deficiency among women in the general population in Japan may not induce significant increase in Cd body burden or Cd-induced tubular dysfunction. PMID- 12719985 TI - Photodynamic therapy for choroidal neovascularization. The Jules Gonin Lecture, Montreux, Switzerland, 1 September 2002. PMID- 12719986 TI - Subretinal organization in stage 5 retinopathy of prematurity. AB - BACKGROUND: The subretinal organization (SRO) seen in patients who undergo vitrectomy for stage 5 retinopathy of prematurity has not previously been characterized. We report our observations of SRO and correlate its development with previous laser and cryotreatment for neovascular disease. METHODS: We surveyed data from 426 eyes in a retrospective chart review of 263 patients that underwent open-sky vitrectomy for stage 5 retinopathy of prematurity. RESULTS: Of 426 eyes evaluated, 130 eyes received laser, cryo, or a combination of both treatments. In 44 eyes (10.3%), SRO was observed and considered the cause of incomplete retinal attachment. Three forms of SRO were identified: subretinal bands (63.6%), subretinal plaques (15.9%), and diffuse SRO (18.2%). One patient had both a band and a plaque. SRO developed in 24 eyes after cryotreatment, 3 after laser, and 2 after combination cryo and laser treatment. Fewer untreated eyes than cryotreatment eyes developed SRO (15 of 296 eyes, 5.1%; ( P=0.0001). Eyes without laser or cryotreatment had a 5.1% frequency of developing SRO. CONCLUSION: Subretinal organization, a previously uncharacterized entity in retinopathy of prematurity, was most frequently identified in the form of subretinal band formation. SRO was identified in 10.3% of all stage 5 eyes evaluated, and was associated with incomplete retinal reattachment in all cases. PMID- 12719987 TI - Clinicopathological correlation of choroidal neovascularization after external beam radiotherapy in age-related macular degeneration. AB - PURPOSE: To analyze the histopathology of choroidal neovascularization after external beam radiotherapy in age-related macular degeneration. METHODS: A retrospective non-case-matched comparative histopathologic study. The histoarchitecture of nine surgically removed subretinal specimens from nine patients that had undergone external beam radiotherapy for exudative age-related macular degeneration was studied. Seven patients had received 20 Gy in 10 fractions and two 15 Gy in 5 fractions with an average time interval between radiotherapy and surgical extraction of 14 months (range 3-28). A consecutive series of classic, mixed and occult choroidal neovascular membranes served as controls. RESULTS: Clinical findings. Radiation-associated choroidal neovasculopathy was angiographically suspected in four patients: a coarse net of vessels on fluorescein angiography developing at the border of previously irradiated choroidal neovascularization was observed in three patients; blebs at the margin of a plaque on indocyanine green angiography were observed in two patients. Pathological findings. Diffuse drusen as well as intra-Bruch's fibrovascular tissue was found in all irradiated specimens. In four specimens an edematous vascularized layer was seen between diffuse drusen and normal-appearing intra-Bruch's fibrovascular tissue. This lesion was not found in the control specimens. A particular correlation for the bleb lesion was not recognized. CONCLUSION: The appearance of an edematous subretinal pigment epithelial vascularized layer between diffuse drusen and normal-appearing fibrovascular tissue in four of nine irradiated membranes may be secondary to previous irradiation. It may correlate with the unusual exudative manifestations observed after external beam radiotherapy. PMID- 12719988 TI - Sequential classification in glaucoma diagnosis. AB - BACKGROUND: Large-scale screening in glaucoma diagnosis is expensive and time consuming. Sequential classification strategies can provide an effective combination of time-efficiency and diagnostic accuracy for glaucoma screening. METHODS: In a cross-sectional clinical study, a sequential diagnostic strategy, based on several psychophysical and electrophysiological tests, was evaluated on measurements from 595 eyes from 310 patients with primary open-angle glaucoma, and 419 eyes from 213 control subjects (age range 18-70 years in each group). Patients and controls successively underwent up to five psychophysical and electrophysiological diagnostic tests. Optic disc morphometry was taken as gold standard. Adapting group sequential techniques, sensitivity and specificity for the whole diagnostic program were controlled, allotting overall error rates of 10%. The criteria for the diagnostic process were developed in a learning sample (677 eyes) and verified in a validation sample (337 eyes). RESULTS: In the validation sample, 62.0% of the examined eyes could be classified, using a sequential 15-min two-step program. An overall 13.6% "gain" of saved time, compared to non-sequential discriminant analysis, was achieved without loss of diagnostic accuracy. A sequential 45-min five-step program classified 68.8% of the whole sample before morphometry, saving approximately 39% of examination time, compared to taking the complete discriminant score. CONCLUSION: Especially in screening, where the use of time-consuming and complicated diagnostic procedures is restricted, the implementation of testing programs based on group sequential strategies might be a promising means of saving personnel resources and reducing inconvenience for patients. PMID- 12719989 TI - Absorption of short wavelengths of endoillumination in indocyanine green solution: implications for internal limiting membrane removal. AB - PURPOSE: To measure the absorption coefficients of short wavelengths, irradiated by a light source used for vitreous surgery, in indocyanine green (ICG) solution and balanced salt solution (BSS), and to determine the implications of these absorption coefficients on retinal phototoxicity caused by ICG-assisted internal limiting membrane removal. METHODS: The absorption coefficients of short wavelengths irradiated by a commercially available light source used for vitreous surgery were measured in ICG solution and BSS using a dual-beam spectroradiometer. RESULTS: The absorption coefficient of wavelengths irradiated by the endoillumination light source in ICG solution was almost the same as that obtained in BSS. The absorbance coefficients of the ICG solutions were 0.18 cm( 1) at 400 nm and 0.03 cm(-1) at 450 nm. In BSS, the absorption coefficients were 0.17 cm(-1) at 400 nm and 0.015 cm(-1) at 450 nm. No significant difference in absorbance was seen between 400 nm and 450 nm ( P>0.05). CONCLUSIONS: The absorption of wavelengths is not greater in ICG solution than in BSS. ICG solution during intravitreal use probably does not enhance retinal photochemical injury during ICG-assisted internal limiting membrane removal. PMID- 12719990 TI - Vitamin E prevents changes in the cornea and conjunctiva due to vitamin A deficiency. AB - PURPOSE: To determine whether vitamin E can prevent changes in the cornea and conjunctiva due to vitamin A deficiency. METHODS: Male infant rats from normal mothers were weaned at 20 days of age and divided into three groups: in group 1, A(-), rats were fed a vitamin A-deficient diet; in group 2, A(-)E(+), rats were fed a vitamin A-deficient diet and injected intraperitoneally with 15 mg of vitamin E per week; group 3 rats were normal controls. At 13 weeks of age, the corneal and conjunctival changes were examined by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). RESULTS: SEM showed more fine microvilli and clear cells in the most superficial epithelial layer of the cornea and conjunctiva in A(-)E(+) rats than in that of A(-) rats. Numerous secretory granules were seen in the goblet cells of the conjunctiva of A(-)E(+) rats, but in A(-) rats there was a total absence of conjunctival goblet cells. TEM showed a severely keratinized corneal surface in A(-) rats. On the other hand, the corneas of A(-)E(+) rats were almost normal in appearance. The conjunctivae of A(-) rats had superficial epithelial keratinization, scarce goblet cells, and many keratohyalin granules in the epithelium and dark cells, while in the conjunctivae of A(-)E(+) rats there were numerous secretory granules in goblet cells and some dark cells. CONCLUSION: The present study suggests that vitamin E has a protective effect against the corneal and conjunctival damage caused by vitamin A deficiency. PMID- 12719991 TI - Lensectomy and vitrectomy decrease the rate of photoreceptor loss in rhodopsin P347L transgenic pigs. AB - BACKGROUND: Photoreceptor degeneration in retinitis pigmentosa (RP) runs an inevitable, gradually progressive course. A wide variety of growth factors of different origins have been shown to slow the rate of degeneration in some rodent models of RP. Recently, lens-derived neurotrophic factors have been shown to rescue degenerating ganglion cells in crush models of the optic nerve. Our objective was to evaluate the potential rescue effect of lensectomy and vitrectomy (L&V) on photoreceptor degeneration in a large-animal model, the rhodopsin P347L transgenic pig. METHODS: We operated on one eye of each of 49 3 week-old pigs--15 vitrectomies and 34 L&V, 6 of which received steroids. Retinal paraffin sections were prepared for all eyes, in addition to immunohistochemistry in four eyes, 8 weeks after L&V. RESULTS: At eight weeks after L&V, operated eyes showed significantly more nuclei in the outer nuclear layer (ONL) than the unoperated fellow eyes. The better preservation of the ONL persisted but was less prominent by 20 weeks after surgery. Steroid treatment did not markedly reduce the better preservation of the ONL seen at 8, 10, and 12 weeks after surgery. The significant difference in cell count between operated and unoperated eyes in the L&V group at 8 weeks was due to the difference in the number of rods, not the cones. CONCLUSION: Lensectomy and vitrectomy delay photoreceptor degeneration in rhodopsin P347L transgenic pigs. Lens-related rescue effect is a probable reason for the delayed degeneration. PMID- 12719992 TI - Heparin drug delivery system for prevention of posterior capsular opacification in rabbit eyes. AB - BACKGROUND: The aim of this study was to investigate the safety and efficacy of a heparin drug delivery system (HEP DDS) for prevention of posterior capsular opacification (PCO) in rabbits. METHODS: Fifty New Zealand albino rabbits (50 eyes) undergoing phacoemulsification were equally divided into five groups receiving normal saline eye drops (group A), a carrier DDS implanted into the posterior chamber (group B), 5% heparin eye drops (group C), a HEP DDS implanted subconjunctivally (group D) and a HEP DDS implanted into the posterior chamber (group E). All the 50 eyes were examined by slit-lamp microscopy. The heparin levels in blood and aqueous humor were measured, and the wet posterior capsules were weighed. RESULTS: All eyes in groups A and B had PCO at 5-7 days, much earlier than in groups C, D and E. Two eyes in group C, three eyes in group D and six eyes in group E showed no signs of PCO throughout the 12-week study. The mean weight of wet posterior capsules from groups A-E was 157.919 mg, 160.091 mg, 81.114 mg, 71.827 mg and 19.984 mg respectively. The mean aqueous humor heparin level in groups C, D and E was 10.279 microg/ml, 13.246 microg/ml and 25.964 microg/ml respectively. Cell proliferation of the posterior capsules in group E was not active. The conjunctiva, cornea, iris, ciliary body and retina remained intact, and no significant toxic reactions were observed. No intraocular hemorrhage occurred during the follow-up. CONCLUSION: Implantation of a HEP DDS into the posterior chamber of experimental animals significantly maintained a higher heparin level in aqueous humor for a relatively long period of time. The findings indicate potential prevention of PCO with minimum toxic and side effects. PMID- 12719993 TI - Long-term alteration in the air-infused rabbit retina. AB - PURPOSE: To investigate the long-term histological changes in rabbit retina damaged by infusion air during vitrectomy. METHODS: Fourteen eyes of 14 rabbits were used. A standard three-port vitrectomy with artificial posterior vitreous detachment followed by fluid-air exchange was performed. After the fluid-air exchange, one side port was kept open for 30 s to freely introduce the air into the vitreous cavity under a pressure of 40 mmHg. Four weeks after surgery, indocyanine green angiography and histological examination were performed. RESULTS: At the air-infused area, indocyanine green angiography revealed the filling delay of the choroidal circulation in the early phase. The retinal surface was smooth and the internal limiting membrane was revealed to be intact by scanning electron microscopy. Light microscopy documented the thinning of the photoreceptor layer in the air-infused area. Transmission electron microscopy showed the disarrangement of the retinal pigment epithelium and loss of choriocapillaries. CONCLUSION: Infusion air during vitrectomy causes long-term outer retinal damage after surgery. Thinning of the photoreceptor cells, disarrangement of the retinal pigment epithelium and loss of choriocapillaries reflect the findings that we have observed previously in clinical cases. Visual field defect is a complication of vitreous surgery using fluid-air exchange. Although the visual field defects were observed just after surgery, abnormal fundus lesions continued to appear over time after the surgical procedure. PMID- 12719994 TI - Elevation of nitric oxide production in human trabecular meshwork by increased pressure. AB - BACKGROUND: The presence of the nitric oxide (NO)-producing enzyme nitric oxide synthase (NOS) in the trabecular meshwork and the effect of various drugs acting through the NO pathway on the outflow facility and trabecular contractility suggest a role for NO in the regulation of outflow and intraocular pressure (IOP). METHODS: To model the effect of elevated IOP on the NO production in the trabecular meshwork, we perfused anterior segments of human donor eyes in vitro and studied the effect of raised perfusion pressure on NO levels in the perfusate. Furthermore, we evaluated using quantitative PCR whether enhanced perfusion pressure had an effect on the NOS gene expression levels. RESULTS: Elevating the pressure from 10 mmHg to 25 mmHg caused a significant increase in NO production from 3.6+/-0.9 pmol/min to 5.9+/-1.6 pmol/min (mean +/- SEM), corresponding to an average increase of 66%. These high NO levels were reduced by application of L-NAME, an NOS inhibitor, to 2.6+/-0.6 pmol/min. Addition of L NAME before raising the pressure decreased the basal NO production but was not able to block the increase in NO production after raising the pressure. The transcript level of iNOS was significantly increased in the trabecular meshwork after raising the perfusion pressure. CONCLUSION: These results show that NO production increased after elevation of the pressure gradient over the trabecular meshwork, accompanied by an upregulation of iNOS gene expression. Previous studies have demonstrated that enhanced NO levels facilitate outflow. Taken together, the data indicate the existence of a regulatory feedback mechanism in the trabecular meshwork, which may contribute to the regulation of the IOP. In this system an increase in IOP will enhance NO production, which, in turn, increases the outflow facility, leading to a normalization of the IOP. PMID- 12719995 TI - AP-1 (c-Fos/c-Jun) is required for corneal epithelial spreading. AB - PURPOSE: We previously demonstrated that the AP-1 components c-fos/c-jun are up regulated in healing rat corneal epithelium in a relatively early phase following epithelial debridement, implicating the AP-1 function in the initiation of cell movement. To explore this hypothesis, we examined the effect of lack of c-Fos and c-Jun protein expressions on the spreading of corneal epithelium and in situ in organ culture. Antisense-oligonucleotide (AS) c-fos-null mice were used for this purpose. METHODS: A rectangular piece of corneal tissue (2 x2 mm) was obtained from each eye of recently killed adult C57BL/6 mice and was incubated for 11 h in culture medium with 8 microM c-fos AS or c-jun AS probe. Sense probes were used for negative control. A rectangular section of corneal tissue was also obtained from each eye of c-fos(-/-), c-fos(+/-) and c-fos(+/+) mice and was organ cultured for 11 h. The length of the path of epithelial spreading on stromal cut surface (both sides) was measured in hematoxylin-eosin-stained specimens. Data were analyzed by unpaired Student's t-test. RESULTS: Addition of c-fos AS to the medium decreased the length of epithelial spread to 40.36% of that in the control with the S probe. Addition of c-jun AS decreased the length of epithelial spreading rate to 42.71% of control with S probe. Lacking c-Fos decreased the epithelial spreading to 17.73% of control data from c-fos(+/-) and c-fos(+/+) mice. CONCLUSION: AP-1 (c-Fos/c-Jun) is required for the corneal epithelial spreading. PMID- 12719996 TI - Expression of AP-1 (c-fos/c-jun) in developing mouse corneal epithelium. AB - BACKGROUND: Activator protein-1 (AP-1) is a ubiquitous transcription factor which is believed to modulate cell behaviors such as proliferation and differentiation during wound healing and embryonic tissue morphogenesis. AP-1 consists of Fos family and Jun family proteins. METHODS: We examined expression pattern of c-fos mRNA and c-Fos protein in developing mouse cornea. Expression of c-jun mRNA and c Jun protein were also examined for comparison. RESULTS: While no c-fos mRNAs were detected on embryonic day (E) 12.5, mRNA for c-fos was detected from E14.5 until postnatal day (P)14. The mRNAs for c-jun were also detected, although the temporal expression patterns differed. c-Fos-immunoreactive nuclei were present from E14.5 through P10 and c-Jun-immunoreactive nuclei were detected from E14.5 through P3. CONCLUSION: These findings indicate that AP-1 ( c-fos/c-jun) transcription factor may play a role in the development and maturation of the corneal epithelium in mice. PMID- 12719997 TI - Toxoplasma gondii retinochoroiditis after cardiac transplantation. AB - PURPOSE: To report 4 cases of Toxoplasma gondii retinochoroiditis in patients having recently undergone cardiac transplantation. METHODS: Review of medical records for 4 patients presenting retinochoroiditis and evidence of T. gondii infection. RESULTS: Patient ranged in age from 25 to 53 years. Ocular symptoms began between 3 and 6 months after transplantation. All patients were under immunosuppressive therapy. Foci of retinochoroiditis were observed unilaterally in three patients and bilaterally in one. Intraocular inflammation was minimal in all cases. Serologic responses were highly suggestive of T. gondii as the etiology in all cases; other causes (CMV retinitis and syphilis) were actively sought and were not found. All patients underwent classic therapy. The three unilateral cases evolved favorably, but the bilateral case, seen late, showed extensive macular scarring. CONCLUSION: Infectious retinochoroiditis is a potentially blinding complication seen after cardiac transplantation, justifying close clinical and serological surveillance or, in certain cases such as mismatched donors, anti-parasitic prophylaxis. PMID- 12719998 TI - Pseudotumor cerebri associated with Sjogren's syndrome. AB - PURPOSE: To report a case of papilledema and pseudotumor cerebri developed in association with Sjogren's syndrome. METHODS: Case-report of a 38-year-old woman with history, imaging and histology confirming the diagnosis of both pseudotumor cerebri and Sjogren's syndrome who presented with bilateral decrease of vision. RESULTS: Papilledema associated with pseudotumor cerebri was observed in both eyes. The patient's visual acuity improved transiently with the administration of intravenous steroids and cyclophosphamide; subsequently she needed a ventriculoperitoneal shunt. CONCLUSION: Sjogren's syndrome should be considered in the different etiologies of pseudotumor cerebri. The major improvement with corticosteroids and ventriculoperitoneal shunt makes prompt diagnosis essential. PMID- 12719999 TI - Is optic nerve head swelling of prognostic value in central retinal vein occlusion? AB - PURPOSE: To investigate the prognostic value of optic nerve head swelling (ONHS) in central retinal vein occlusion (CRVO) and compare it to other prognostic factors. METHODS: Seventy-four patients with CRVO were studied retrospectively. The parameters analysed were the initial presence of ONHS, the fluorescein angiographic appearance, the implicit time in the 30-Hz flicker ERG and the initial visual acuity. The aspects of outcome studied were the development of neovascular complications and the visual acuity 1 year after the thrombotic event. RESULTS: Fluorescein angiography, ERG and initial visual acuity were of prognostic value in CRVO, whereas ONHS was of questionable value. CONCLUSION: ONHS is most likely of no prognostic value in CRVO. PMID- 12720000 TI - Reconstruction of the radionuclide spectrum of liquid radioactive waste released into the Techa river in 1949-1951. AB - The major part of the liquid radioactive waste released by the Mayak Production Association (PA) radiochemical plant into the Techa river occurred in 1949-1951, but there is information on only one single radiochemical analysis of a water sample taken on 24 and 25 September 1951. These data are here used to assess the spectrum of radionuclides that were released between 1949 and 1951. For this purpose, details of the radiochemical methods of radionuclide extraction and radiometric measurements of beta-activity used at Mayak PA in the 1950s have been taken into account. It is concluded that the data from the radiochemical measurements agree with the theoretical composition of fission products in uranium after exposure times in the reactor (120 days) and subsequent hold times (35 days) that were typical for the procedures at that time. The results of the analysis are at variance with assumptions that underlie the current Techa river dosimetry system. They confirm the conclusion that the external doses to the Techa river residents in the critical period up to 1952 were predominantly due to short-lived fission products. PMID- 12720001 TI - A study of 110mAg in aquatic and terrestrial ecosystems. AB - Experiments on a simulated terrestrial agricultural ecosystem were carried out using the pot culture approach. The most representative plants in local vegetable gardens were selected to investigate the root uptake of (110m)Ag. The results show that carrot, kale and flowering cabbage have the largest transfer factor values among the vegetables. Flowering cabbage, as the most popular leafy vegetable in Hong Kong and the South China area, can be used as a biomonitor for radioisotope contamination in vegetables. Soil column and adsorption tests were also carried out to study the leaching ability of the silver isotope in soil and (110m)Ag was mainly adsorbed in the top 1 cm of soil regardless of the pH value. Experiments on a simulated aquatic ecosystem for freshwater fish and marine organisms were carried out in glass aquaria. The freshwater fish Cyprinus carpio, the marine fish Cuvier and some local abundant seashore molluscs were selected to investigate the kinetic metabolism of (110m)Ag in the compartmental system. The results show that molluscs absorb (110m)Ag much more than fish. Clibanarius infraspinatus has the largest concentration factor among the marine organisms selected. Fish liver, although representing a minor portion of the total body mass, shows the highest (110m)Ag concentration factor, whereas muscle, although representing a major portion of the total body mass, is characterized by an absence of (110m)Ag. PMID- 12720002 TI - Induction of micronuclei in human fibroblasts and keratinocytes by 25 kV x-rays. AB - A relative biological effectiveness (RBE) not much larger than unity is usually assumed for soft x-rays (up to approximately 50 keV) that are applied in diagnostic radiology such as mammography, in conventional radiotherapy and in novel radiotherapy approaches such as x-ray phototherapy. On the other hand, there have been recent claims of an RBE of more than 3 for mammography and respective conventional x-rays. Detailed data on the RBE of soft x-rays, however, are scarce. The aim of the present study was to determine the effect of low energy x-rays on chromosomal damage in vitro, in terms of micronucleus induction. Experiments were performed with 25 kV x-rays and a 200 kV x-ray reference source. The studies were carried out on primary human epidermal keratinocytes (HEKn), human fibroblasts (HFIB) and NIH/3T3 mouse fibroblasts. Micronucleus (MN) induction was assayed after in vitro irradiation with doses ranging from 1 to 5.2 Gy. Compared to the effect of 200 kV x-rays, 25 kV x-rays resulted in moderately increased chromosomal damage in all cell lines studied. This increase was observed for the percentage of binucleated (BN) cells with micronuclei as well as for the number of micronuclei per BN cell. Moreover, the increased number of micronuclei per micronucleated BN cell in human keratinocytes and 3T3 mouse fibroblasts suggests that soft x-rays induce a different quality of damage. For all cell lines studied the analysis of micronucleus induction by 25 kV soft x rays compared to 200 kV x-rays resulted in an RBE value of about 1.3. This indicates a somewhat enhanced potential of soft x-rays for induction of genetic effects. PMID- 12720003 TI - A track structure model for simulation of strand breaks in plasmid DNA after heavy ion irradiation. AB - We present a track structure model based on the local dose deposited around heavy ion tracks to explain the cross sections for single-strand and double-strand break induction in plasmid DNA in different aqueous buffers. The model is based only on measurable quantities, namely the effect distribution for inducing strand breaks after x-ray irradiation as a function of dose, and the radial dose distribution of the heavy ion track. The effect of indirect DNA damage mediated by free radicals produced in the water surrounding the DNA is accounted for by allowing the radial dose distribution to be smeared in space by an effective target size corresponding to the squared sum of the geometrical extension of the plasmid molecule and the mean free drift path of the radicals in the buffer solution. Our calculations reproduce well the measured cross sections for single strand and double-strand break induction in SV40 plasmid DNA in various buffer solutions both as a function of the LET and of the specific energy of the heavy ion. PMID- 12720004 TI - Poznan's program of cochlear and brainstem implantation: a general review. AB - The cochlear implantation program in Poznan started in January 1994. Within 8 years, 151 patients were implanted (149 cochlear and 2 auditory brain-stem implants). The implanted group comprised 111 children and 40 adults. We have extended the lower age limit for cochlear implant candidates, and the youngest implanted patient was 18-months-old at the time of surgery. We have also implanted six multi-handicapped subjects. All patients were implanted with different nucleus devices (22, 24 M, 24 K, 24 Contour, 24 Double Array and 24 ABI). Insertion of the Nucleus 24 Contour electrode carrier requires a little larger cochleostomy in comparison with conventional straight array. After insertion, the intraoperative measurements are taken (impedance telemetry, electrically evoked stapedius reflex and NRT). The day after surgery, each patient undergoes an X-ray examination (the cochleogram or cochlear view). The calculated length of the electrode array insertion was 12.75-24.0 mm (mean: 22.05 mm) and the degree of electrode rotation was 180-540 degrees (mean: 292.82 degrees). After speech processor switch-on, we performed NRT measurements. Our results indicated that the NRT threshold was in 51.4% of the dynamic range within the group of examined patients. The adults' progress of rehabilitation was checked during individual sessions. The rehabilitation of children included two rehabilitation schedules: continuous ones and weekly rehabilitation camps. The progress of the recovery process of implanted children enabled them to change their school profile from hearing impaired to mainstream. Voice rehabilitation is often monitored by acoustic analysis using the Multi Dimensional Voice Program (MDVP). PMID- 12720005 TI - Risk of variant Creutzfeldt-Jakob disease and tonsillectomy. PMID- 12720006 TI - Ischemic optic neuropathy after endoscopic sinus surgery: a case report. AB - For the past 2 decades, endoscopic sinus surgery (ESS) has proven effective for treating paranasal sinus disease. Orbital complications of varying degrees, from mild orbital hematoma to catastrophic blindness, have been widely reported. However, defects of the visual field resulting from post-ESS ischemic optic neuropathy (ION) has not to our knowledge been reported in the literature. We were presented with a 51-year-old male patient suffering from loss of sight following an otherwise uneventful ESS. ION is a rare condition, characterized by acute or subacute postoperative loss of sight. The major risks for developing ION include intraoperative anemia, hypotension and systemic illnesses such as hypertension, diabetes or renal failure. Otorhinolaryngologists should be aware that this condition may occur following an uncomplicated ESS procedure, and patients should be given prompt opthalmological consultation when loss of sight is diagnosed postoperatively. Early aggressive and rapid correction of blood pressure and blood transfusions may be helpful in the treatment of patients who develop ION after surgery. PMID- 12720007 TI - Posterior epistaxis: endonasal exposure and occlusion of the branches of the sphenopalatine artery. AB - Intractable posterior epistaxis (PE) is a frequent emergency for which different treatment modalities are available. While nasal packing causes extreme discomfort and angiography with consecutive selective embolization is not available everywhere, recent studies emphasize the value of sphenopalatine artery (SPA) occlusion by different techniques and indicate success rates of 13-33%. In our institution, previously endoscopic management of PE consisted either of isolated coagulation of an identified bleeding source (group A) or cutting and coagulation of arterial branches running through the sphenopalatine foramen (SPF) (group B). According to our neuroradiological and rhinological experience we developed a modification of SPA transsection and coagulation following identification of the division in conchal and septal branches of the SPA (group C). During a 26-month period the success rates of these three techniques in 95 patients were compared prospectively. The three modalities revealed a re-bleeding rate of 3 out of 21 (21%) in group A, 1 in 6 (16.7%) in group B and 3 in 69 (4.3%) in group C. At the level of the SPF, 36 out of 69 patients had one conchal branch, whereas 30 (43.5%) had two and 3 (4.4%) had three. If SPA transsection and coagulation for intractable PE is adopted the anatomic varieties of the SPA with its division in conchal and septal branches have to be taken into account. According to our experience the septal branch of the SPA plays a major role in PE. Its occlusion significantly improves the success rate of PE treatment. PMID- 12720008 TI - Ethanol and acetone stimulate the proliferation of HaCaT keratinocytes: the possible role of alcohol in exacerbating psoriasis. AB - Alcohol has been reported to be a risk factor in psoriasis mainly based on the observation that there is a higher prevalence of alcohol abuse in individuals with psoriasis. The mechanism by which alcohol affects this disease is still elusive. So far there are no reports describing the effects of metabolites relevant to alcohol metabolism on the growth of human keratinocytes. In the present study we examined the effects of ethanol and acetone, which exceeds its normal endogenous level in the blood of heavy drinkers, on the proliferation of HaCaT keratinocytes. HaCaT cells were incubated for 30 min in the presence of various concentrations of ethanol (2.14 m M-1.71 M) and acetone (1.7 mM-1.36 M). The numbers of viable and proliferating cells were determined at different times after ethanol and acetone treatment. The effects of ethanol and acetone on the mRNA levels of genes characteristic for proliferating keratinocytes such as alpha5 integrin, keratinocyte growth factor receptor and cyclin D1 were studied by reverse-transcriptase polymerase chain reaction. Both ethanol and acetone induced proliferation of HaCaT cells. The maximum increase in the number of viable cells and the maximum proliferative response was observed with 4.28 m M ethanol and 13.6 m M acetone. The alpha5 integrin, keratinocyte growth factor receptor and cyclin D1 mRNA levels were higher compared to the controls as early as 2 h after ethanol and 30 min after acetone treatment of the cells. The stimulatory effect of ethanol and acetone on human keratinocytes may be one of the reasons why psoriasis can be precipitated by alcohol misuse. PMID- 12720009 TI - The significance of subacromial arthrography to verify partial bursal-side rotator cuff ruptures. AB - BACKGROUND: The purpose of this study was to verify a partial bursa-side rupture of the rotator cuff (RC) using different imaging techniques with special emphasis on the validity of a specific method of subacromial arthrography (SAA). METHODS: Patients (n=92, mean age 53.8 years) with a subacromial impingement syndrome underwent sonography, magnetic resonance imaging (MRI), and SAA. All diagnostic results were checked by subsequent arthroscopic surgery. RESULTS: Of 31 surgically verified ruptures, 17 showed a partial rupture located towards the bursa. These had been detected by MRI and ultrasound with a sensitivity of 64% and 41%, respectively, while SAA as a diagnostic tool yielded a sensitivity of 82%. In 14 cases of complete RC ruptures, all imaging techniques had a similar sensitivity of 86-93%. CONCLUSION: It appears that SAA is an adequate and valid diagnostic tool for the detection of partial bursa-sided RC ruptures. Ultrasound and MRI showed a comparably lower sensitivity. It is therefore concluded that SAA has clear advantages in the diagnosis of this defect with the consequence that open surgical techniques can be avoided. PMID- 12720010 TI - Management of infected shoulder replacement. AB - BACKGROUND: The purpose of the present work is to present treatment options and our own results for patients with infected shoulder alloarthroplasties. METHODS: Twelve patients with an infected shoulder replacement were treated. Their age ranged from 56 to 82 years. Indications for surgical revision were clinical symptoms compatible with an infection or positive serologic tests, especially an elevated C-reactive protein. Aspirated intra-articular fluid with a white blood cell count above 30,000 or positive for bacterial growth was also an important diagnostic feature. Retrospective analysis differentiated three groups with three different treatment regimens. Group 1 with early infection but without soft tissue involvement (n=1): this particular patient underwent arthroscopic synovectomy. Group 2 with early infection and soft-tissue involvement within 4 weeks after index surgery (n=1): this patient underwent open synovectomy. The largest group was group 3 with late infection (n=10): these patients were treated with two-stage revision and a temporary spacer. RESULTS: The time between explantation and reimplantation ranged between 4 weeks and 6 months. With the temporary spacer, an anatomically stable condition could be established for all patients, and reconstruction of the humeral length even in long implants was possible. All patients underwent physiotherapy with the temporary spacer in place. A positive intraoperative microbiologic specimen was only found in 4 patients. In both group 1 and 2 patients, the infection healed, and thus the original implant could be kept in situ. In 8 patients, the temporary spacer was removed and exchanged for a regular implant. The postoperative raw Constant score at the time of the last follow-up examination was 48, due mainly to a loss of motion and power. All shoulders were stable, and the elbow function was good. CONCLUSION: Use of an antibiotic-loaded spacer allows successful treatment of infected shoulder replacements. PMID- 12720011 TI - Isolated, closed rupture of the common femoral artery resulting from blunt trauma. AB - We present a case of a 13-year-old child with an isolated, closed rupture of the common femoral artery and no associated bony injury sustained as a result of blunt trauma. Though few isolated cases of femoral artery injury secondary to blunt trauma have been reported in the literature, most of them were the result of a bony/penetrating injury. The probable mechanism of injury is discussed. PMID- 12720012 TI - Patellar tendon rupture and marked joint instability after total knee arthroplasty. AB - BACKGROUND: Patellar tendon rupture is a rare complication of total knee arthroplasty (TKA). Multiple repair methods have been described in the literature. METHODS: A 66-year-old woman suffered a patellar tendon re-rupture and marked joint instability within 6 months after revision TKA. She underwent re revision TKA and extensor mechanism reconstruction with femoral quadriceps tendon and augmentation by a Leeds-Keio ligament. RESULT: It was fairly difficult to acquire a satisfactory range of motion as well as gain in knee extension capacity by eliminating the extension lag. CONCLUSION: Patellar tendon ruptures after TKA should be repaired as soon as they are recognized. PMID- 12720013 TI - Improving reliability in the classification of fractures of the acetabulum. AB - BACKGROUND: Plain radiographs of the pelvis are routinely used in the initial assessment of patients with suspected fractures of the acetabulum. It is necessary for orthopaedic resident trainees, emergency physicians as well as orthopaedic surgeons who infrequently treat trauma patients to be able to describe these fracture patterns reliably to traumatologist orthopaedic surgeons who ultimately take over the patient care. Our purpose was two-fold: (1) to determine the reliability of the component parts of the Letournel classification of acetabular fractures involving six anteroposterior (AP) radiographic lines, and (2) to examine whether the addition of oblique radiograph views (Judet views) would improve the reliability. METHODS: Thirty sets of AP and oblique radiographs (Judet views) of the pelvis were selected from a hospital database to represent various types of acetabular fractures. Six reviewers (three orthopaedic trainees and three community orthopaedic surgeons) independently reviewed the radiographs. For each radiograph, the reviewer classified the acetabular fracture according to the Letournel classification. In addition, each reviewer utilized a simplified classification scheme using six radiographic lines on the AP pelvic radiograph. Interobserver reliabilities among reviewers were reported along with the intraclass correlation coefficient (ICC) and kappa values. RESULTS: Agreement for the Letournel classification increased with increasing physician experience (trainees ICC=-0.14 and community surgeons ICC=0.56). Interobserver reliability between trainees and community surgeons improved when the six radiographic lines were used (range kappa=0.09-0.89). The oblique pelvic radiographs (Judet views) did not significantly improve reliability among physicians. CONCLUSIONS: In this study we report the following: (1) the reliability of the Letournel classification improves with level of training, (2) physicians with less experience with acetabular fractures have significantly better agreement in identifying fractures using the six radiographic lines on the AP film than the Letournel classification, and (3) agreement among the reviewers for the AP pelvic radiograph is not improved with additional oblique (Judet) views. PMID- 12720014 TI - Lateral J-plate fixation in comminuted intercondylar fracture of the humerus. AB - BACKGROUND: Anatomical reduction and rigid internal fixation of the articular surface is key for functional recovery in types C2 and C3 (AO classification) fractures of the distal humerus. Since these fractures are associated with varying degrees of comminution in both the medial and lateral columns, rigid fixation of both columns is necessary for early mobilization of the elbow joint. METHODS: We performed a prospective study of the lateral J-plating technique in 17 patients with comminuted intercondylar fracture of the humerus. There were 9 type C2 and 8 type C3 fractures by the AO/ASIF classification. A lateral J-plate was applied along the lateral aspect of lateral column, and the lag screws were inserted into the medial column through the plate hole. A medial reconstruction plate was selectively applied on the posterior aspect of medial column. RESULTS: All fractures united by 18.7 weeks on average (range 10-28 weeks). Seven patients presented with limitation of motion of the elbow joint, and 5 of them underwent arthrolysis and excision of the heterotopic ossification. The final range of motion of the elbow joint was flexion contracture of 16 deg to further flexion of 121 deg on average. Functional results were excellent in 9 cases, good in 6, fair in 1, and poor in 1 according to Murray's criteria. CONCLUSION: Lateral J-plating with selective augmentation of the medial column provided sufficient rigidity and good clinical results. PMID- 12720015 TI - Magnetic resonance imaging features of epidermoid cyst in the extremities. AB - BACKGROUND: Epidermoid cyst is a common benign soft-tissue tumor that occurs in the skin. There have been very few descriptions about the magnetic resonance (MR) images of epidermoid cysts in the extremities. MR imaging findings of epidermoid cysts may be characteristic enough to make the preoperative diagnosis possible. The purpose of this paper is to elucidate the MR imaging features of epidermoid cysts arising in the limbs. METHODS: We evaluated the MR images of five patients with an epidermoid cyst in the limbs. MR imaging and other radiological examinations were evaluated. T1-weighted (T1W), T2-weighted (T2W), and Gd enhanced T1W images were taken in all the patients. RESULTS: The size of the lesion ranged from 2 to 10 cm with an average of 5.2 cm. All the tumors were located in the subcutaneous tissue with a well-circumscribed margin. On T1W MR imaging, the tumor had a slightly high signal intensity in three out of five cases and iso-signal intensity in the remaining two cases. On T2W images, they all showed high signal intensity. Irregular low signal intensity areas were noted in the tumors on both T1W and T2W images. No enhancement was observed inside the tumors. CONCLUSION: Subcutaneous tumor location, well-circumscribed margin, and isosignal to slightly high signal intensity with some heterogeneity on T1W MR images and high signal intensity T2W MR images, and no enhancement are useful features for a diagnosis of epidermoid cyst of the extremity. PMID- 12720016 TI - Arthroscopic management of tibial plateau fractures--comparison with open reduction method. AB - BACKGROUND: Arthroscopic treatment of tibial plateau fractures were compared with open traditional methods retrospectively. METHODS: Twenty-eight patients were included in this comparative study based on fracture patterns and divided into two groups. Using Schatzker's classification system, patients with type II (split depression), type III (isolated depression) fractures, or other fracture types were included. Nineteen of these patients were treated by arthroscopically assisted management (group S), and the remaining 9 underwent the conventional open method (group O). RESULTS: There was no significant difference between both groups in terms of duration of operation, postoperative flexion, and clinical results. In group S, however, the postoperative rehabilitation was easier and faster (the time to obtain 120 degrees of flexion was 4.6+/-3.9 weeks in group S, 9.1+/-8.6 weeks in group O). Furthermore, 16 of 19 patients (84%) in group S obtained an anatomical reduction (defined as <2 mm of residual displacement after surgery), whereas in group O, only 5 of 9 patients (55%) had an anatomical reduction. Also, accurate diagnosis and treatment of any associated joint pathology was possible with arthroscopic management. CONCLUSION: The arthroscopic procedure is recommended in selected tibial plateau fractures. PMID- 12720018 TI - [The efforts for longevity: what maximum lifespan does society bear?]. PMID- 12720019 TI - [Increasing life expectancy--what are the promises of demography?]. AB - The prospects for a longer life are improving, for life expectancy is growing continuously. In Germany new-born boys are looking forward to 74.4 years, girls even to 80.6 years of life. But Germany is only mid-range internationally. As there is a linear trend for 160 years, it seems realistic that life expectancy in top countries will be 100 years in 2060. If we look more thoroughly we find that there are important differences in Germany. Whereas the southern parts are in the pole position, the former East Germany is at the bottom of the league. This separation is apparently caused by the different political systems in postwar Germany. There are even more differences if smaller regions are examined. The reasons for these differences are due to many influences, including personal features, conditions of life, lifestyle and, of course, medical treatment. As these factors are partly determined by individual behavior, people are partly responsible for their life and therefore also for the further development of life expectancy. PMID- 12720020 TI - [Increasing life expectancy--the big social issue of the 21st century?]. AB - Due to the combined effects of a persistent low birth rate, the forthcoming retirement of the "baby-boom generation" and an increasing life expectancy, the German system of "social security" will face severe challenges in the coming decades. The foreseeable doubling of the ratio between pensioners and employees by 2050 would cause a strong increase in social security contributions and taxes for the "pay-as-you-go"-financed pension system. In addition, the expected tripling of highly aged people would increase the public expenditures for health care and nursing care of elder people, which also have to be financed by the future working generations. In order to lessen the increasing burden on future generations, social reforms are necessary soon. If these are not performed, the distributional conflict between young and old people could become worse over the forthcoming decades. PMID- 12720021 TI - [A long life--a wish and its limits. Aging with dignity and meaning]. AB - In the journal "Gerontologie" (1991, Oswald WD et al. Eds.), the article on "Langlebigkeit" (longevity) is followed by the article on "Lebenszufriedenheit" (satisfaction in life) for purely alphabetical reasons. However, there is a deeper meaning in this order which refers to a connection that should not be neglected. "The number of our days is 70 years. Or maybe 80 years if..." To have a long and satisfying life is a deeply rooted wish in the human mind. Medicine and modern, improved conditions of living let this come true for many. However, what undoubtedly is a value by itself and has been praised as success until recently, now causes personal and social problems. We are dealing with a covert problem in particular: Which decisions that concern us and are likely not to please us, are others going to take when we are old? "I need a coat already. For me the sun is already declining". The wish to live as long as possible is always connected with hope, that is hope to have a fulfilling life. However, there are limits to that hope due to the constancy of maximal life expectation, multiple morbidity related with old age and consequent shortening of life. At the same time basic existential experiences as uniqueness, loneliness and the limits of the human mind increase with age. So what does it mean to fill life in old age with meaning and dignity? Nowadays dignity is constantly talked about. That is possibly why it is on the retreat. Dignity is understood as something that can be ascribed and if that is so, we can simply abandon it since theology is not a known discipline any more. Theology and religion in general could bring about a new understanding of the meaning of life as it could relate something incomprehensible with a potential interpretation. Life in old age also comprises the personal and private wherein the memories of the personal life attain a particularly important function. PMID- 12720022 TI - [Everlasting solidarity? Longevity and generational relations]. AB - Demographic development of industrial societies is determined by continuous low birth rates and increasing life expectancy. The dramatic change in generational structure will be an enormous challenge not only for the public social security systems; as an original place of inter-generational relations, family is particularly faced with increasing life expectancy and the chances and risks of longevity. Need for nursing care is such a risk of longevity, realizing that only about 3% aged between 60 and 80 are in need of care, but about 25% at the age of 80 or older. The majority of nursing cases is well-cared at home, mostly by relatives. Although one might care for relatives for many reasons--not only for love and altruism--empirical data insist on indicating strong family ties based on senses of solidarity and commitment. The focal point of the following article is a case review describing highly emotional family relationships in a home care situation. On the one hand, the description clarifies family solidarity representing the basis of German long-term care insurance since 1995; on the other hand it demonstrates the enormous burden caring for an old mother or father. Increasing life expectancy might even intensify burdens, so that fewer and fewer relatives might be able or willing to accept them. In this matter, new mixtures of private and public services will attain increasing importance. Considering German long-term care insurance a pioneer, it expresses a new sense of limited public responsibility assuming the existence of sufficiently strong family ties. PMID- 12720023 TI - [From alms to entitlements. A historical approach on private and public attitudes towards long life expectancies]. AB - At all times in history, there have been people reaching a high age. However, long life expectancies as a--relatively seen--socially non-stratified phenomenon are of a very recent date. This essay identifies three different currents which have shaped the related massive development of population. First, there was what could be called the "democratization of long life expectancies", rooted in the steadily growing access to the fruits of medical and technical progress during the past centuries. Second, the state constantly took over from the private sector responsibilities to provide services for old people. Third and simultaneously, a commonly shared idea of entitlement to a period of retirement after the working career gained acceptance. These three currents are summarized in the essay under the heading of the development of "social rights". Until the end of the 19th century those rights were totally neglected. It was only under the impression of industrialization, the appearance of a strong labor movement, world economic crisis and the catastrophe of two world wars that a conscience for the social dimension of citizenship developed. The history of German social policies from Bismarck to the present day serves as a measurement for these developments towards social rights. In this respect, particular attention is paid to the "Grosse Rentenreform" of 1957, the breakthrough of the modern German welfare state. PMID- 12720024 TI - The medical management of primary open-angle glaucoma in the elderly patient. AB - Glaucoma is a frequent cause of blindness in the elderly. It is defined as a neuropathy of the optic nerve for which the main risk factor is an elevated intraocular pressure. The management of glaucoma is described with an emphasis on diagnostic procedure and on the topical drug treatment. The potential side effects of these drugs are discussed from the geriatric point of view. PMID- 12720025 TI - [How stable is the attitude of aged people towards life-extending measures? Results of a 3-year follow-up in nursing home residents]. AB - OBJECTIVE: To explore the stability of attitude towards life-extending measures in aged people. METHOD: Face-to-face interviews of nursing home residents at an interval of 3 years (prospective longitudinal survey). RESULTS: During the first interview period in 1997 (n=50), 19 nursing home residents (38%) had advocated treatment with antibiotics in a hypothetical scenario of acute life-threatening pneumonia. 15 individuals (30%) had refused such treatment, while statements from a further 16 (32%) had been ambivalent. Three years later, 19 individuals could be re-interviewed (28 had died in the meantime, 3 suffered from advanced dementia). 16 of these expressed the same or a similar attitude towards the above scenario as they did three years earlier (correspondence 84.2%, p<0.01). CONCLUSION: The results indicate a high stability of attitude towards life extending measures in aged people. This finding may have implications for the meaning of advance directives in geriatric long-term care. PMID- 12720026 TI - [The influence of socio-demographic factors on personality dimensions in the elderly]. AB - The personality factors neuroticism, extraversion, openness for experiences, agreeableness and conscientiousness were investigated in a random population sample of n=497 people over 60 years old with the help of a German version of the NEO Five-Factor Inventory - NEO-FFI (2, 6). The results were compared with those from n=497 people between 18 and 30 years old and with those from n=1051 people between 31 and 60 years old. The elderly described themselves as being less extroverted, less open, more agreeable and more conscientious than the people under 60 years old. Means of the personality factors were not associated with age in the over 60 years old. The average level of neuroticism and conscientiousness was significantly higher in women over 60 years old than in men. People living in the former Eastern Germany were more open and conscientiousness than those living in former Western Germany. A higher level of openness was found in people with a higher educational level. There was a higher level of neuroticism in people living without a partner than in those living with a partner. PMID- 12720027 TI - [The role of depression in rehabilitation of geriatric patients with hip fracture]. AB - The effect of depression on the rehabilitation course of 64 geriatric inpatients with hip fracture was examined. At the beginning of treatment 38.5% had a depressive disorder according to GDS-15. After 3 weeks it was still 32%, although in 7 patients the GDS-score had decreased. Additionally, 18% had a cognitive disorder (MMSE<24). There was a correlation between depression and Barthel-Index at the start of treatment (58.4; SD 14.7 vs 48.3; SD 21.8; ANOVA; p<0.01), Barthel-Index at discharge (86.1; SD 13.1 vs 75.0; SD 25.11; ANOVA; p<0.01), mobility measured by timed "Up and Go" (Chi(2); p<0.01) and the number of prescribed drugs, as equivalent for severity of somatic diseases. Treatment of depression was inadequate in the beginning and at follow-up. Treated patients had a better outcome than untreated. PMID- 12720028 TI - [Lung embolism in the elderly]. AB - Elderly people have an increased risk of suffering from thrombo-embolic events. Lung embolism is often found as a cause of death in autopsies in elderly people. However, it is unknown whether the increased age is a risk factor per se or whether elderly people suffer more often from co-morbidities associated with a higher thrombo-embolic risk. Immobility is the most important risk factor for thrombo-embolic events. Other risk factors include trauma, surgery, venous stase, genetic factors (thrombophilia) and a history of past thrombo-embolic events. Although the probability of suffering from a thrombo-embolic event increases with age, it can be difficult to find the correct diagnosis in elderly people: symptoms like dyspnoe or chest pain might be explained by other cardio-pulmonary diseases. In this review, we would like to give an overview about the diagnostic and therapeutic steps in elderly people. PMID- 12720029 TI - The effects of war on children in Africa. AB - There is no doubt that the effects of war extend to the most vulnerable members of society, including children. Although armed conflicts occur throughout the world, the African continent seems to be a particular background for civil and international wars. The aim of this study was to identify causes of conflict in Africa and to evaluate the effect of war on children and their health in order to make practical recommendations to health care workers dealing with children in the setting of war. All articles written in the past 5 years concerning "war" and "children" were identified by means of a literature search and internet review. Contrary to common belief, the causes of conflict are complicated and multi factorial. The effects of war on childhood are disastrous and include severe negative effects on general paediatric health status. Short-term recommendations for health care workers working with children in war include supply of emergency medical infrastructures, basic health care, rehabilitation and education. Long term recommendations include orchestrating the relief and support efforts from both national governments and international non-profit organisations and speeding up of economic recovery. The causes of conflict in Africa are complex and unlikely to be resolved soon. The effects of war on children are horrendous in many ways, but can be limited by providing timely and appropriate health care. PMID- 12720030 TI - Universal distribution of c-kit-positive cells in different types of Hirschsprung's disease. AB - Interstitial cells of Cajal (ICCs) have been reported to play the role of a pacemaker in regulating bowel motility. The relationship between neurons and ICCs, however, remains unclear. Hirschsprung's disease (HD) is an ideal model for investigating this relationship. The operated specimens obtained from 6 short and 3 long segment aganglionosis patients and 3 controls were used as the subject materials in this study. ICCs were immunohistochemically identified using a specific antiserum c-kit, a tyrosine kinase receptor expressing ICCs. Nitrergic nerves were demonstrated by NADPH-diaphorase (NADPH-d) histochemistry. C-kit immunohistochemistry was also combined with protein gene product 9.5 (PGP 9.5; as a general neuronal marker). In the normoganglionic segment of HD, numerous c-kit positive cells and NADPH-d positive neurons were found in the proper muscle layer, including Auerbach's plexus. In the oligoganglionic segment, the number of c-kit-positive cells and NADPH-d neurons slightly decreased. In the inner border of the circular muscle layer (IBCM), the c-kit-positive cell networks and NADPH-d activities remained in short segment cases, while both of them were absent in the long segment cases. In the aganglionic segment, c-kit positive cells were present universally but the number of them was slightly decreased in the proper muscle layer. The c-kit-positive cell networks of IBCM were seen where extrinsic neurons were present, while they were almost completely absent where extrinsic neurons were absent in the proximal zone of the long segment cases. C-kit positive cells were present universally in the oligoganglionic as well as aganglionic segments of HD. The distribution and properties of c-kit positive cells were related to the presence of extrinsic neurons in aganglionic segment. Based on these findings, aperistalsis is considered not to relate with c-kit positive cells, and c-kit positive cells are not supposed to have a neurogenic origin and can develop without neurons, however the lack of enteric neurons may influence the full differentiation of ICCs. PMID- 12720031 TI - Infantile myofibromatosis of the central nervous system. AB - BACKGROUND: Infantile myofibromatosis is the most common fibrous disorder of infancy and childhood. It may occur in two distinct forms: multicentric and solitary. In both cases involvement of the central nervous system (CNS) is unusual: brain myofibromas are usually intracranial in proximity of the dura mater, with infiltration of the calvarial bones and secondary brain compression. Spine myofibromas are exceptional and most of the cases reported in the literature represent secondary locations of visceral lesions. The natural history of myofibromas of the CNS in infantile myofibromatosis is characterized by a period of rapid growth, subsequent stabilization, and spontaneous regression in many cases. On these grounds surgical treatment is reserved for lesions that compress the brain or spinal cord and show an obvious progression. Small lesions or tumor residuals in asymptomatic children without visceral involvement should be closely observed as first choice. CASE REPORT: We report the case of a 10 month-old child with a solitary intramedullary myofibroma, extended from C7 to the conus. The patient underwent partial surgical removal of this lesion. Control MRI has shown the spontaneous slow reduction of the tumor residual and a progressive improvement of preoperative neurological conditions has been observed. To our knowledge this is the first case in which a solitary myofibroma was localized within the spinal cord. PMID- 12720032 TI - Orientation tuning of motion-sensitive neurons shaped by vertical-horizontal network interactions. AB - We measured the orientation tuning of two neurons of the fly lobula plate (H1 and H2 cells) sensitive to horizontal image motion. Our results show that H1 and H2 cells are sensitive to vertical motion, too. Their response depended on the position of the vertically moving stimuli within their receptive field. Stimulation within the frontal receptive field produced an asymmetric response: upward motion left the H1/H2 spike frequency nearly unaltered while downward motion increased the spike frequency to about 40% of their maximum responses to horizontal motion. In the lateral parts of their receptive fields, no such asymmetry in the responses to vertical image motion was found. Since downward motion is known to be the preferred direction of neurons of the vertical system in the lobula plate, we analyzed possible interactions between vertical system cells and H1 and H2 cells. Depolarizing current injection into the most frontal vertical system cell (VS1) led to an increased spike frequency, hyperpolarizing current injection to a decreased spike frequency in both H1 and H2 cells. Apart from VS1, no other vertical system cell (VS2-8) had any detectable influence on either H1 or H2 cells. The connectivity of VS1 and H1/H2 is also shown to influence the response properties of both centrifugal horizontal cells in the contralateral lobula plate, which are known to be postsynaptic to the H1 and H2 cells. The vCH cell receives additional input from the contralateral VS2-3 cells via the spiking interneuron V1. PMID- 12720033 TI - Pharmacological properties of l-glutamate receptors associated with the crayfish hindgut. AB - Pharmacological agents were used to characterize glutamate receptors associated with crayfish hindgut. l-Glutamate reliably increased tonus in isolated hindguts of Procambarus clarkii and suppressed spontaneous hindgut contractions at concentrations of 10 micromol l(-1) or higher. Quisqualate and ibotenate mimicked the effects of L-glutamate. Experiments with strips and rings of hindgut tissue indicate that glutamate acts on both circular and longitudinal muscles. Hindgut contractions were not affected by (+/-)-alpha-amino-3-hydroxy-5-methyl-isoxazole 4-propionoic acid, N-methyl- d-aspartate, domoate or the metabotropic glutamate receptor agonist, (1S,3R)-1-amino-1-carboxycyclopentane-3-acetic acid. Picrotoxin, at 50 micromol l(-1), did not alter the ibotenate-induced reduction in contraction frequency, suggesting that this effect is not produced by inhibitory glutamate receptors. The glutamate-induced increase in tonus was antagonized by Joro spider toxin, JSTX-3. Thus, glutamate receptors associated with crayfish hindgut muscles are of the quisqualate type but are also sensitive to ibotenate. Elevating extracellular potassium concentration mimicked all of the effects of glutamate, suggesting that excessive depolarization may contribute to the suppression of contractions at high agonist concentrations. PMID- 12720034 TI - Crop scents affect the occurrence of trophallaxis among forager honeybees. AB - Previous evidence indicates that the recognition of the nectar delivered by forager honeybees within the colony may have been a primitive method of communication on food resources. Thus, the association between scent and reward that nectar foragers establish while they collect on a given flower species should be retrieved during trophallaxis, i.e., the transfer of liquid food by mouth, and, accordingly, foraging experience could affect the occurrence of these interactions inside the nest. We used experimental arenas to analyze how crop scents carried by donor bees affect trophallaxis among foragers, i.e., donors and receivers, which differ in their foraging experience. Results showed that whenever the foragers had collected unscented sugar solution from a feeder the presence of scents in the solution carried by donors did not affect the occurrence of trophallaxis nor its dynamics. In contrast, whenever the foragers had previous olfactory information, new scents present in the crop of the donors negatively affected the occurrence, but not the dynamics of trophallaxis. Thus, the association learned at the food source seems to be retrieved during trophallaxis, and it is possible that known scents present in the mouthparts of nest-mates may operate as a triggering stimulus to elicit trophallactic behavior within the hive. PMID- 12720035 TI - To stay or not to stay: the contribution of tactile and thermal cues to coming to rest in newborn rabbits. AB - Newborn rabbits, like other altricial mammals, demonstrate thermotaxis and when placed on a thermal gradient locate and come to rest at physiologically appropriate temperatures. Little is known, however, about the sensory-motor components contributing to the in energetic terms important decision of the young to cease locomotion and come to rest. We investigated the behavior of newborn rabbits on two thermal gradients; linear in which pups could use tactile cues from the arena wall, and concentric in which pups were unable to use such cues. On both gradients pups located the warm, thermal-neutral area within the 200-s test time, thereby demonstrating their ability to orient appropriately using thermal cues alone. Unexpectedly, however, pups on the concentric gradient failed, or took significantly longer, to come to rest than pups on the linear gradient. Since the speed of locomotion of pups on the linear gradient was significantly slowed when they were in contact with the arena wall, and in most cases they came to rest in contact with it, we suggest that not only thermal but also tactile cues may be important in bringing young mammals to rest in a thermally appropriate environment. PMID- 12720036 TI - Robotic assisted microsurgical vasal reconstruction in a model system. AB - Our objective was to determine whether or not male reproductive microsurgery is adaptable to current robotic technology. We devised a model vas deferens system using rat vasa deferentia and completed vasal anastomosis with full-thickness and mucosal robotically placed, Sharpoint 10-0 bicurve nylon sutures. Experienced and inexperienced microsurgeons performed separate anastomoses. Both groups of microsurgeons completed anastomoses with accuracy and enhanced comfort. The robotic graspers had the dexterity to delicately handle the 10-0 sutures and needles. We conclude that through our model vas deferens system, we demonstrated the feasibility of applying robotic technology to male reproductive microsurgery. This pilot study reveals some advantages of the robot and suggests future use of this system in a new urological application domain. PMID- 12720037 TI - Herbal and complementary medicine in chronic prostatitis. AB - Chronic prostatitis is a very common and poorly understood condition with significant impact on quality of life. The etiology of prostatitis can be multifactorial and can present with a variety of symptoms. Given the lack of proven efficacy of conventional therapies such as antibiotics, many patients have turned to phytotherapy and other alternative treatments. This review will cover the alternative therapies commonly used in prostatitis with an emphasis on those with published data. These treatments include phytotherapy (quercetin, bee pollen) and physical therapy. Complementary therapies have shown the potential to help men with prostatitis, particularly when allopathic therapies have failed. PMID- 12720040 TI - The iliac angle: a sonographic marker of trisomy 21 during the midtrimester: dependency of fetal lying? AB - The aim of this study was to evaluate the efficiency of the fetal iliac wing angle measurements in the detection of trisomy 21 during the second trimester and the impact of the fetal position on this measurement. During a 43-month period, the iliac wing angle was prospectively measured in 695 fetuses at genetic ultrasonography performed before amniocentesis. The iliac wing angle measurements were performed in a true axial section of the fetal pelvis and the relative position of the fetal spine was recorded (decubitus, lateral). Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and likelihood ratio (LR) were computed for multiple cutoff angles. The prevalence of trisomy 21 was 1.4%; 447 fetuses were in lateral position, 240 fetuses were in decubitus position, including, respectively, 2 and 8 fetuses with trisomy 21 (8 fetuses with other chromosomal anomalies were excluded). In euploid fetuses, the mean iliac wing angle was 83.7 degrees in decubitus and 68.7 degrees in lateral position; in fetuses with trisomy 21 the respective mean angles were 104.9 degrees and 102.5 degrees. A receiver operating characteristics (ROC) curve provided an absolute cutoff angle of 90 degrees, predicting trisomy 21 with a sensitivity of 100%, a specificity of 72.4%, an NPV of 100%, a PPV value of 11.1%, and an LR of 3.6. The measurement of iliac wing angles in an axial section of the fetal pelvis is dependent on spine orientation and provides an efficient cutoff angle for the exclusion of trisomy 21 in patients at risk. PMID- 12720042 TI - Progressive systemic sclerosis-polymyositis overlap syndrome with eosinophilic pleural effusion. AB - Pleural fluid rarely occurs in patients with progressive systemic sclerosis (PSS) or polymyositis (PM) with no lesions in the pulmonary area. Pleural fluids in patients with autoimmune diseases are mostly dominated by monocytes and lymphocytes but very rarely contain increased eosinophils. We report a 55-year old male with PSS-PM overlap syndrome and eosinophilic pleural effusion. Air invasion into the pleural cavity and the antituberculous therapy could be ruled out as causes for the patient's eosinophilic pleural effusion, because the differential eosinophil count was already as high as 19% from the first thoracentesis before the start of antituberculous therapy. Infections and malignant tumor also were unlikely causes based upon the negative pleural fluid results and the negative pleural biopsy findings, except for nonspecific inflammation. After the administration of corticosteroid, the pleural effusion decreased promptly, with normalization of serum creatine phosphokinase and C reactive protein concentrations. PMID- 12720043 TI - Septic arthritis caused by Granulicatella adiacens: diagnosis by inoculation of synovial fluid into blood culture bottles. AB - Granulicatella species, formerly known as nutritionally variant streptococci, cause a variety of infections, primarily endocarditis. We report the first culture-proven case of a Granulicatella species causing septic arthritis. A 68 year-old female presented with knee pain and swelling. She was initially evaluated with arthrocentesis and arthroscopy, but no organism was identified. Her pain improved after a brief course of antibiotics but recurred 3 months later. She underwent repeat arthrocentesis, with direct inoculation of synovial fluid into blood culture bottles. Granulicatella adiacens was recovered from both bottles. She was treated with cefazolin for 4 weeks combined with gentamicin for the first 2 weeks. Her knee pain and swelling resolved without evidence of recurrence. Granulicatella should be considered in cases of septic arthritis with initially negative synovial fluid cultures. Inoculation of blood cultures bottles with synovial fluid may increase the diagnostic yield for these species. PMID- 12720044 TI - Early-onset hemochromatic arthropathy in a patient with idiopathic hypermobility syndrome. AB - Hemochromatosis is a genetic disease related to human leukocyte antigen (HLA) A3, B7, and B14 histocompatability antigens resulting in increased iron absorption from the gastrointestinal tract and deposition of iron in tissues. Arthropathy is not uncommon in the late stage of disease. Characteristic radiologic findings are commonly observed in the wrists and metacarpophalangeal joints as well as the hips, knees, and ankle joints. Presented here is a 34-year-old male with hemochromatosis and bilateral shoulder, knee, and ankle pain. Radiologic examination revealed osteoarthritic findings in both ankle joints and chondrocalcinosis in the knee joints. All the major criteria of hypermobility syndrome were observed on physical examination. The early-onset arthropathy seen with this hemochromatosis is thought to result from hypermobility syndrome. PMID- 12720045 TI - Anemia, serum vitamin B12, and folic acid in patients with rheumatoid arthritis, psoriatic arthritis, and systemic lupus erythematosus. AB - OBJECTIVE: Although anemia is frequent in inflammatory rheumatic diseases, data regarding vitamin B12 status is scarce. The purpose of this study was to analyze the incidence and nature of B12 and folic acid (FA) deficiencies in a cohort of rheumatic patients with rheumatoid arthritis (RA), psoriatic arthritis (PsA), and systemic lupus erythematosus (SLE). METHODS: Levels of B12, FA, and parameters of anemia were recovered or examined in 276 outpatients. In those with recent findings of low serum B12 levels, further studies of serum homocysteine (Hcy) and urine methylmalonic acid (MMA) levels were performed. RESULTS: The incidence of anemia was high: 49%, 46%, and 35%, in RA, SLE, and PsA, respectively. Low levels of serum B12 were also frequent (24%), with almost similar occurrence in the three disease groups. Deficiency in FA was rare (<5%). Mean levels of both vitamins did not differ significantly among the three groups. No correlation between serum B12 levels and anemia was found. In the 15 patients with recently detected low B12 levels, Hcy and MMA were evaluated before and following B12 therapy. In ten of them, baseline Hcy levels were high, while MMA was increased in one patient only. Response to B12 administration, i.e., a decrease in Hcy and/or MMA levels, was noticed in four patients only, suggesting that only 26% of the low-serum-B12 patients had true B12 deficiency. CONCLUSIONS: The incidences of anemia and decreased serum B12 levels were high in these three groups of rheumatic patients. However, true tissue deficiency seems to be much rarer. PMID- 12720046 TI - Signal transduction pathways regulating osteoclast differentiation and function. PMID- 12720047 TI - Analytical and clinical evaluation of the Bio-Rad HPLC kit for measurement of type I collagen cross links. AB - The measurement of hydroxylysylpyridinoline (PYD) and lysylpyridinoline (DPD), the degradation products of type I collagen, by manual HPLC assay posed practical difficulties. The present study was undertaken to evaluate the first commercially available HPLC kit, which provides a convenient substitute for cumbersome classical HPLC methods. The HPLC procedure is based on an improved sample clean up chromatography, convenient ready-to-use HPLC reagents, and quicker isocratic elution of PYR and DPD on reverse-phase analytical column. The analytical parameters assessed include sensitivity, within- and between-assay variation, method comparison, recoveries, and interference. Clinical evaluation included discriminatory power of PYD and DPD and response to treatment of osteoporosis patients with Alendronate. DPD and PYD concentrations showed linear ( r(2) > 0.99) response between 10-400 pmol/ml and 75-4000 pmol/ml, respectively. The average within-assay imprecision, over a range of clinically relevant cross-links concentrations, was CV < 7% ( n = 30). The total imprecision ( n = 35 days), by ANOVA, for PYD and DPD was CV < 7.5% and CV < 10%, respectively. Average spike recovery was 95.4% +/- 6.5%. Comparison with the historical HPLC method exhibited a close correlation ( r values between 0.87 and 0.91, P < 0.0001). Creatinine corrected DPD in postmenopausal ( Z score = 2.4, P < 0.05, n = 17) and osteoporotic ( Z score = 3.0, P < 0.01, n = 29) women were 44% and 64% higher, respectively, compared to premenopausal samples ( n = 15). Similarly, PYD concentration was 26% and 54% higher in postmenopausal and postmenopausal osteoporotic women, respectively. There was a 47% ( P < 0.001) decrease in DPD concentration ( n = 16), and a 30% decrease in PYD concentration after 90 days of treatment of osteoporotic patients with Alendronate. DPD concentration correlated with N-telopeptide with an r value of 0.69 ( n = 67, P < 0.0001). The reported kit method is substantially simpler and precise than the manual method. DPD concentrations determined by the current method reaffirm the clinical value in identifying increased bone resorption in pathological conditions and monitoring response to antiresorptive therapy. PMID- 12720048 TI - Localization and inhibitory effect of basic fibroblast growth factor on chondrogenesis in cultured mouse mandibular condyle. AB - The condylar cartilage, an important growth site in the mandible, shows characteristic modes of growth and differentiation, unlike the limb bud cartilage. To elucidate the mechanism of chondrogenesis at the condylar cartilage, we analyzed the effects of basic fibroblast growth factor (bFGF) on the growth and development of mouse mandibular condyle using serum-free organ culture and on the expression of genes related to the chondrogenesis. Further, we investigated the localization of bFGF in cultured condyle by immunohistochemistry. The present immunohistochemical observations showed that bFGF is localized in the extracellular matrix of the mesenchymal condylar anlage, the perichondrium and the proliferative cell zone, and that immunostaining was diminished in the metachromatically stained area. In the condyle culture with added recombinant human bFGF (rhbFGF) for 5 days, the area occupied by hypertrophic chondrocytes in the mandibular condylar cartilage was reduced. A reverse transcription-polymerase chain reaction (RT-PCR) assay also showed that the mRNA levels of aggrecan and type X collagen were reduced compared with nontreated tissues. Treatment with rhbFGF for 2 days decreased cell proliferation in the perichondrium, and bFGF downregulated the Indian hedgehog (Ihh), parathyroid hormone-related protein (PTHrP), bone morphogenetic protein 4 (BMP4), and core-binding factor alpha1 (Cbfa1) expression in the RT-PCR assay. These findings suggest that bFGF has the ability for inhibitory regulation of condylar growth, via the inhibition of proliferation and differentiation of chondrocytes, and that this inhibitory regulation is related to the downregulation of growth factors and transcription factors. PMID- 12720050 TI - Vitamin K(2) (menaquinone 4) reduces serum undercarboxylated osteocalcin level as early as 2 weeks in elderly women with established osteoporosis. AB - Twenty elderly osteoporotic women with vertebral fracture(s) were randomly allocated to two groups; women in the MK(4) group received calcium with menaquinone 4 (MK(4)) at a dose of 45 mg/day for 2 weeks, and women in the control group received calcium alone for the same period. Serum intact osteocalcin (OC) and undercarboxylated osteocalcin (uc-OC) levels were measured by immunoradiometric assay and enzyme immunoassay, respectively, at baseline and on the 7th and 14th days following the start of the treatment. There were no differences in the baseline data including age, weight, phylloquinone, menaquinone 4, menaquinone 7, OC, and uc-OC levels between the MK(4) group and the control group. Administration of MK(4) significantly raised the MK(4) level from 0.20 +/- 0.10 (mean +/- SE) pg/ml to 15.09 +/- 5.62 pg/ml ( P < 0.04), and reduced serum uc-OC levels from 2.80 +/- 0.93 ng/ml to 1.76 +/- 0.56 ng/ml ( P < 0.05) at the end of the study, respectively. No significant changes in these levels were observed in the control group. Serum OC levels were stable during the period in both groups. In this randomized prospective study, the MK(4) group shows a reduction in the serum uc-OC level within 2 weeks without any significant change in OC, suggesting that the uc-OC is changed to carboxylated OC. This early effect of MK(4) on bone metabolism may be estimated by the measurement of serum uc-OC in elderly osteoporotic women with vertebral fractures. PMID- 12720049 TI - Combination use of vitamin K(2) further increases bone volume and ameliorates extremely low turnover bone induced by bisphosphonate therapy in tail-suspension rats. AB - Bisphosphonate is a potent inhibitor of bone resorption, which results in the increase of bone volume. However, bisphosphonate treatment may lead to extremely low bone turnover and abnormal bone microstructure. In this study, we examined whether the combination of bisphosphonate with vitamin K(2) treatment may have beneficial effects on bone turnover and trabecular microstructure as well as on bone volume loss by using tail-suspension model rats. In these model rats, bone mineral density (BMD) decreased with histological evidence of enhanced bone resorption and suppressed bone formation. By bisphosphonate treatment, BMD was increased compared with that of tail-suspended rats. Osteoclast surface per bone surface (Oc.S/BS) and number of osteoclasts per bone perimeter (N.Oc/B.Pm) were reduced and mineral apposition rate (MAR) decreased, suggesting extreme suppression of bone turnover. However, trabecular structure examined by microfocus CT was apparently abnormal. By contrast, combination of bisphosphonate with vitamin K(2) leads to further increase of bone volume. MAR and BFR as well as Oc.S/BS and N.Oc/B.Pm were increased compared with those of the bisphosphonate treated group. However, abnormal structure of trabeculae in secondary spongiosa was not completely ameliorated. These data suggested that concomitant use of vitamin K(2) with bisphosphonate excessively ameliorates too much suppression of bone turnover while more efficiently preventing bone volume loss. PMID- 12720051 TI - Decrease in serum leptin by troglitazone is associated with preventing bone loss in type 2 diabetic patients. AB - The thiazolidinedione (TZD) class of antidiabetic drugs has been shown to inhibit the formation of bone-resorbing osteoclasts in vitro and to decrease bone resorption markers in vivo. These drugs also inhibit the expression of leptin in adipocytes. Less leptin can be associated with higher bone mass, based on analyses of mice deficient in leptin action. Effects of 1-year treatment with troglitazone, a member of the TZDs, on bone mineral density (BMD) and bone metabolism were examined in 25 Japanese type 2 diabetic patients. Glucose metabolism was improved, whereas body mass index and percent body fat did not change throughout the study. The percent change of BMD was negatively correlated with that of serum leptin, whereas it was not associated with changes of bone metabolic markers, type I collagen N-telopeptide (NTx), bone alkaline phosphatase (ALP), body mass index, or HbA1c. Serum leptin decreased in 68% of subjects (responders) after 1-month treatment and was consistently lower than the basal level throughout the treatment. Percent changes of BMD were significantly higher in the responders than in the nonresponders and in nondiabetic subjects at 6 and 12 months. NTx and bone ALP decreased at 1 month but increased thereafter in either group of patients. Thus, it is suggested that the decrease in serum leptin with no reduction in body fat mass by troglitazone is associated with preventing bone loss in type 2 diabetic patients. Hence, TZDs may have an advantage for diabetic patients who have risk factors for osteoporosis. PMID- 12720052 TI - Analysis of linear growth in survivors of childhood acute lymphoblastic leukemia. AB - Therapy for childhood acute lymphoblastic leukemia (ALL) is entering a new era in terms of quality-of-life. In the current study, 21 patients with childhood-onset ALL were assessed for linear growth, bone mineral density (BMD), and endocrinological status, focusing especially on longitudinal analysis of the growth of each patient. Linear growth was uniformly attenuated during therapy in all patients. In contrast, after the cessation of therapy, the growth of each patient varied widely from attenuated to dramatic catch-up growth. In pubertal survivors who had received chemotherapy and cranial irradiation during prepuberty, the degree of growth after the cessation of therapy was negatively correlated with changes in height Z scores during therapy ( r = -0.76, P= 0.004). One of the factors involved in catch-up growth, urinary N-telopeptide/creatinine (U-NTx/Cr), was significantly higher in patients whose Z scores decreased after cessation of therapy ( P = 0.01), despite normal pubertal development and normal endocrinological assessments. The present study revealed individual differences in linear growth after the cessation of therapy and suggests the importance of catch-up growth during puberty. PMID- 12720053 TI - Relationship of upper body fat distribution to higher regional lean mass and bone mineral density. AB - We investigated the relationship of upper body fat distribution to regional bone mineral density (BMD). Subjects were 361 regularly menstruating premenopausal women with right-side dominance. Age, height, and weight were recorded. Regional (arms, L2-L4 of the lumbar spine, and legs) BMD, regional (arms, trunk, and legs) lean mass, and the ratio of trunk fat mass to leg fat mass amount (trunk-leg fat ratio) were measured by dual-energy X-ray absorptiometry. Body fat distribution was assessed by trunk-leg fat ratio. Trunk-leg fat ratio was positively correlated with regional BMD ( r = 0.120-0.216; P < 0.05- P < 0.001) and regional lean mass ( r = 0.162-0.229; P < 0.01- P < 0.001). Regional lean mass was more positively correlated with regional BMD ( r = 0.319-0.475; P < 0.001). These relationships remained significant after adjusting for age and height. However, correlation of trunk-leg fat ratio with regional BMD disappeared after additionally adjusting for regional lean mass. In premenopausal women, upper body fat distribution is associated with higher regional BMD via greater regional lean mass irrespective of age and height. PMID- 12720056 TI - Abstracts of the 31st Annual Meeting of the North American Society for Psychosocial Obstetrics and Gynecology. February 19-22, 2003, Lihue, Hawaii, USA. PMID- 12720054 TI - Establishment and evaluation of bone mineral density reference databases appropriate for diagnosis and evaluation of osteoporosis in Chinese women. AB - This study was designed to establish Bone Mineral Density (BMD) Reference Databases for multiple skeletal sites appropriate for the diagnosis and evaluation of osteoporosis (OP) in Chinese women. We recruited 2702 healthy Chinese women, 5-96 years of age, for BMD assessment. BMD values at multiple skeletal sites including anteroposterior (AP) and lateral (Lat) lumbar spine, hip, and forearm were measured by dual-energy X-ray absorptiometry (DXA) using a QDR 4500A device; results were analyzed according to age group using eight regression models. BMD Reference Databases (CWD) were established according to the best regression equation and compared with Hologic reference databases for "Oriental Women" (OWD). Results indicated that the cubic regression model was superior to the quadratic, linear, logarithmic, and exponential regression models, etc. for our purpose, with a determinate coefficient ( R(2)) of 0.363 0.650 ( P = 0.000). We included 1636 female patients, aged 35-86 years, in our tests. In comparison with Hologic Reference Databases, the mean detection rate of OP in the newly established BMD Reference Databases for Chinese Women (CWD) was 16.0% +/- 2.68% lower (range, 13.7%-20.5%) at the AP spine, 16.8% +/- 11.0% lower (range, 3.5%-32.8%) at the Lat spine (except for L4), 18.7% +/- 4.6% lower (range, 12.6%-24.2%) at the hip, and 14.3% +/- 6.9% higher (range, 4.7%-24.3%) at the forearm. The difference in detection rates for OP was significant between the two reference databases ( P = 0.000), which was consistent with the differences in peak BMD values and the biological variability between them. Based upon our data, we confirmed that the Hologic BMD Reference Databases for Oriental Women (OWD) were not suitable for the diagnosis of OP in Chinese women; the BMD Reference Databases for Chinese Women (CWD) established in this study would provide reliable diagnostic standards for detection of OP in the women of South China. PMID- 12720058 TI - Psychoneuroendocrine correlates of secondary amenorrhea. AB - Functional hypothalamic amenorrhea is a common, non-organic and theoretically reversible form of anovulation due to reduced hypothalamic GnRH drive. Numerous studies suggest that this altered hypothalamic homeostasis is caused by a synergism between psychogenic challenge, promoted in part by dysfunctional attitudes, and metabolic compromise induced by undernutrition and overexercise. The recent growing interest in psychiatric comorbidity underlines the importance of reconsidering the boundaries between psychological disorders and somatic conditions. That not withstanding, it is mandatory in gynecological endocrinology to explore the issue of secondary amenorrhea from a psychoneuroendocrine perspective in order to devise biopsychosocial interventions which address the individual distress. In this brief review we will try to critically discuss the issue providing evidences from personal studies as clues for better understanding the extent of the complex psychoneuroendocrine network controlling menstrual function. PMID- 12720059 TI - Retrospective evaluation of the treatment for breast cancer: how does the patient's personal experience of the treatment affect later adjustment to the illness? AB - The aim of the present study was the evaluation of patients' treatment experience for breast cancer and its possible associations with the illness adjustment process. To examine perception and experience of treatment during the diagnostic and the inpatient phase, as well as during chemo-and radiotherapy and during after-care, a self-compiled questionnaire was employed. To assess illness adjustment, the German version of the Hospital Anxiety and Depression Scale, the "Freiburger Fragebogen zur Krankheitsverarbeitung" and the "Veranderungsfragebogen des Erlebens und Verhaltens" were used. 126 patients participated in the study, all diagnosed for breast cancer during the past 5 years. We found an association between negative experience of all treatment phases (doctor-patient relationship, communication) and impaired adjustment process reflected by high anxiety and depression scores and more problematic coping strategies. PMID- 12720060 TI - Somatization disorders in obstetrics and gynecology. AB - Somatization refers to the phenomenon of physical symptoms not being conditioned by physical structural defects but primarily by mental processes. The basic understanding integrating psychodynamic, cognitive behavioral and systemic views conceptualized the underlying process as a signal processing circle in which the perceptual, interpretative and the effective-behavioral part interact in a dysfunctional manner. The clinical entities of this basic mechanism are variable in the localization and intensity of the symptoms which is reflected in the existing classification of ICD 10 and DSM IV. These classifications do not differentiate between gender. In obstetrics and gynecology somatization disorders are quantitatively important and qualitatively distinct. The pathogenetic factors are linked to gender specific biopsychosocial conditions, psychoendocrine interactions and genital organ subjective experiences. Therefore we propose a gender-sensitive classification into three groups of somatization disorders: Somatizing reactions during specific life phases or transitional periods, pychoendocrine disorders and somatoform disorders of the female genital organs. In each group the dysfunction may manifest itself mainly on the perceptual part or the effective-behavioral part of the signal processing circle. The pathogenetic pathways, comorbidities, specific diagnostic and therapeutic interventions for these different groups are summarized. PMID- 12720061 TI - Gender and perversion--what constitutes a "bad mother". AB - BACKGROUND: There is a need for reconsidering the conceptualisations of female perverse behavior, especially in connection with motherhood. METHODS AND FINDINGS: Based on case material obtained through psychoanalytic psychotherapy with female patients from a psychosomatic gynecological outpatient clinic, the characteristics of the psychic structure of these patients who presented symptoms of deliberate self harm and of misusing and mistreating their children, are outlined. Another common trait is the embeddedness of their perverse behavior in a generational chain of transmission. Female patients who mistreat their children had been victims of traumatising experiences in their own biography, inflicted by their mothers and directed towards their bodies. DISCUSSION: Female perverse behavior, therefore, is fundamentally different from male perversion: the perverse act in women is aimed against themselves and/or their children. Currently used diagnostic statistical manuals lack categories to describe this symptomatology adequately. Further research is requested to understand a mother's perverse actions and thus develop treatment strategies, without marginalizing these patients. PMID- 12720062 TI - Occupational conditions exceed the importance of non-occupational conditions and ill health in explaining future unemployment among women and men. AB - BACKGROUND: Unemployment is relatively common today, but the importance of work characteristics in relation to future unemployment has seldom been studied. METHODS: The relation between occupational and non-occupational factors, as well as ill health, in 1993 and unemployment in 1997 has been examined using an exploratory approach. Both analyses of associations and cluster analysis have been performed. RESULTS: Reduced psychological well-being and some occupational factors mainly related to job insecurity and, among women, also few opportunities for development at work--were found to be risk indicators for unemployment later on. To some extent, also earlier unemployment periods predicted unemployment in 1997. The influence of reduced psychological well-being and earlier unemployment periods on the associations between occupational factors in 1993 and 1997 consisted mainly of a decrease of the influence of job insecurity. Seven clusters were identified. One of them was male dominated, where the level of unemployment was very high and the individuals had reduced psychological well-being, insecure working conditions and demanding living conditions. The female equivalent was characterised by a high level of unemployment, multi-demands at work and poor musculoskeletal health. CONCLUSIONS: Reduced psychological well-being and job insecurity were risk indicators for later unemployment among both genders, as were also few opportunities for development at work among women. These factors predicted unemployment, also when age and other non-occupational factors were taken into consideration. The cluster analysis identified groups of individuals with highly demanding occupational and non-occupational conditions in 1993 and a high level of unemployment in 1997. PMID- 12720063 TI - Premenstrual symptomatology, locus of control, anxiety and depression in women with normal menstrual cycles. AB - The present study was designed to investigate the relationships between premenstrual symptomatology, locus of control, anxiety, and depression in women with normal menstrual cycles. Sixty-nine female participants completed a survey, comprised of the Menstrual Distress Questionnaire (MDQ; Moos, 1968), Levenson's (1981) locus of control scales, the Depression Anxiety Stress Scale (DASS; Lovibond and Lovibond, 1995), and a questionnaire constructed by the researchers based on the DSM-IV criteria for Premenstrual Dysphoric Disorder (PMDD). Both overall and specific subtypes of premenstrual symptomatology were found to correlate with external locus of control, anxiety, and depression. In addition, locus of control was found to moderate the relationship between premenstrual symptomatology, anxiety and depression. Finally, women who were in the premenstrual phase when completing the questionnaire scored significantly lower on the internal scale than those in either the follicular or early luteal phases. It was concluded that an external locus of control may be associated with a susceptibility to depression or anxiety when certain premenstrual or postmenstrual changes are experienced. PMID- 12720064 TI - Localization of cerebral lesions in aphasia--a computer aided comparison between men and women. AB - It is still a matter of debate whether there are differences between men and women concerning the localization of higher cerebral functions. To further elucidate this problem we conjointly evaluated the aphasia protocols and corresponding computer-assisted tomography (CT) scans of 49 men and 35 women who presented with unilateral ischemic cerebral lesions. Both, the aphasia tests and CT scans, were fed into computer programs warranting a high degree of objectivity. Comparing the four main aphasic syndromes we found differences for each of them displaying a more posterior extension of lesions with global and amnestic aphasia in men and larger lesions with Broca and Wernicke aphasia in women. When all templates were compared, men displayed a wider extension within the left hemisphere than women whose lesions were more focused in the perisylvian area. This study confirms earlier findings claiming that aphasic men and women differ regarding the anterior-posterior extension and the frequency of lesions within the classical language zones. However, while the predominance of men's lesions in the posterior areas was in accordance with previous findings, the greater overall scatter within the left hemisphere contradicts published studies. PMID- 12720065 TI - A double-blind, placebo-controlled trial of fluvoxamine in binge eating disorder: a high placebo response. AB - Twenty subjects with binge eating disorder were randomly assigned to flexible dose fluvoxamine or placebo for 12 weeks. A significant reduction in binge frequency, Beck Depression Inventory scores and the eating concern, shape concern and weight concern subscales of the Eating Disorder Examination were noted for both fluvoxamine (n = 9) and placebo (n = 11) groups. There were no significant differences between fluvoxamine and placebo for any treatment outcome variables. The findings from this small trial contribute to the inconsistent results of antidepressant studies in binge eating disorder. PMID- 12720069 TI - Unethical research funding contracts: just say NO! PMID- 12720068 TI - Influence of graded facetectomy and laminectomy on spinal biomechanics. AB - Facetectomy and laminectomy are techniques for decompressing lumbosacral spinal stenosis. Resections of posterior bony or ligamentous parts normally lead to a decrease in stability. The degree of instability depends on the extent of resection, the loading situation and the condition of the intervertebral discs. The correlation between these parameters is not well understood. In order to investigate how these parameters relate to one another, a three-dimensional, non linear finite element model of the lumbosacral spine was created. Intersegmental rotations, intradiscal pressures, stresses, strains and forces in the facet joints were calculated while simulating an intact spine as well as different extents of resection (left and bilateral hemifacetectomy, hemilaminectomy and bilateral laminectomy, two-level laminectomy), disc conditions (intact and degenerated) and loading situations (pure moment loads, standing and forward bending). The results of the modelling showed that a unilateral hemifacetectomy increases intersegmental rotation for the loading situation of axial rotation. Expanding the resection to bilateral hemifacetectomy increases intersegmental rotation even more, while further resection up to a bilateral laminectomy has only a minor additional effect. Hemilaminectomy and laminectomy only differ in their effect for ventriflexion and muscle-supported forward bending. Two-level laminectomy increases the intersegmental rotation only for standing. Degenerated discs result in smaller intersegmental rotations and higher disc stresses at the respective levels. Decompression procedures affect the examined biomechanical parameters less markedly in degenerated than in intact discs. Resection of posterior bony or ligamentous elements has a stronger influence on the amount than on the distribution of stresses and deformations in a disc. It has only a minor effect on the biomechanical behaviour of the adjacent region. Spinal stability is decreased after a laminectomy for forward bending, and after a two level laminectomy for standing. For axial rotation, spinal stability is decreased even after a hemifacetectomy. Patients should therefore avoid excessive axial rotation after such a treatment. PMID- 12720070 TI - Resource utilization and cost of episodes of febrile neutropenia in children with acute leukemias and lymphomas. AB - The resource utilization and cost of 51 episodes of febrile neutropenia in children with leukemia and lymphomas who were admitted to the Pediatric Oncology Institute (GRAAC) of the Federal University of Sao Paulo were analyzed. Patients aged 60 days to 21 years with confirmed diagnoses of acute myeloid leukemia, acute lymphoid leukemia, non-Hodgkin lymphoma, or Hodgkins disease who presented axillary temperature above 38 degrees C at least once episode, or between 37.5 degrees C and 38 degrees C on three occasions during a 24-h period, neutrophil count below 500/mm(3), or between 500/mm(3)and 1,000/mm(3) but expected to fall below 500/mm(3) were included in the study. The patients' ages varied between 1 and 15.6 years, and 67% of the patients were male. The median cost per treated episode was US dollars 2,660 (2,039). Hospitalization costs accounted for 62% of the total cost of the treatment, antibacterials accounting for 23%. Episodes in patients with documented infections had a higher median direct cost than episodes in patients with fever of unknown origin (P=0.018). There was a trend for a higher median direct cost in episodes among patients with a worse prognostic factor, such as type of underlying disease, presence of documented infection, and longer duration of neutropenia. This is the first study to evaluate the economics of febrile neutropenia episodes in Brazil, and serves as a basis for resource utilization and costs incurred in the treatment of such patients in this country. PMID- 12720071 TI - Creating a language for "spiritual pain": why not to speak and think in terms of "spiritual suffering"? PMID- 12720072 TI - Distress of inpatients with terminal cancer in Japanese palliative care units: from the viewpoint of spirituality. AB - A qualitative study was conducted in an attempt to improve our understanding of the spiritual distress of terminally ill cancer patients in Japan. The subjects were inpatients at four approved palliative care units in Japan. The patients were interviewed and they expressed their own experiences in which spirituality was considered to be identifiable. Literal records of the interviews were analyzed using constant comparative analyses as proposed in relation to the grounded theory approach. The analysis included 11 inpatients and a variety of expressions of distress were obtained. After the conditions of the subjects were evaluated in order to search for common factors related to distress, consciousness of the gap between the patient's aspirations and the present situation were found to cause gap-induced distress. Distress was classified into three categories: distress due to the gap between the present situation and how the individual wanted to live, how the individual wished to die, and the individual's wish to maintain relations with others. The aspirations causing the gap were then interpreted from the viewpoint of spirituality as "anchors in life" for Japanese patients with terminal cancer. It was also revealed that in patients who possessed pictures of how they wished to die as their "anchors in life" and who were in a severe physical condition, distress increased and they became confused after their physical symptoms were relieved following admission to PCU. PMID- 12720073 TI - Antidepressant prescribing in community cancer care. AB - GOALS: To describe patterns of antidepressant (ADs) prescribing in community oncology practice. PATIENTS AND METHODS: Data were collected using an electronic medical record on all staged breast, colon, and lung cancer patients in three community-based oncology practices. The data were analyzed retrospectively, using descriptive and bivariate analyses and multivariate logistic regression modeling. There were 850 breast, 299 colon, and 473 lung cancer patients identified in this analysis. MAIN RESULTS: Overall, 19.2% of breast, 11% of colon, and 13.7% of lung cancer patients had been prescribed ADs during the 2-year period. The clinic in which cancer treatment was received predicted AD prescribing. The relationship between AD administration and age proved to be nonlinear; the pattern exhibited an "inverted U" shape. Patients with comorbidities and on pain medications were more likely to be administered ADs. Colon cancer patients on pain medications were five times more likely to be administered ADs than those not on pain medications. CONCLUSIONS: While some predictors of AD prescribing appear to be consistent with other studies, such as being on pain medication, there is still a great amount of variability in prescribing patterns across community practices, age groups, and cancer diagnoses. This study demonstrates that prescriptions of ADs seem to be influenced by parameters other than psychopathology. Given the importance of major depression in oncology care, diagnosis of psychiatric disorders and prescription patterns of psychotropics should be part of the routine monitoring and quality management in oncology patient care. PMID- 12720074 TI - The health-related quality-of-life impact of histamine dihydrochloride plus interleukin-2 compared with interleukin-2 alone in patients with metastatic melanoma. AB - The aim of this study was to compare the quality-of-life (QOL) effects of interleukin-2 (IL-2) alone with those of IL-2 plus histamine dihydrochloride in the setting of a multicenter, randomized trial for patients with metastatic melanoma. QOL data were collected from July 1997 to March 2000 during a phase III trial comparing subcutaneous histamine plus IL-2 and IL-2 alone. Prior to each treatment cycle, patients completed the 76-item Quality-of-Well-Being Scale-Self Administered (QWB-SA) questionnaire, the Overall State of Health (OSH) item, and the General Health Perception (GHP) item. A longitudinal data analysis using the generalized estimating equations approach was performed to compare changes in QWB SA scores over time between treatment groups, and predicted QWB-SA scores from the regression analysis were used to calculate quality-adjusted survival duration over the 12-month study period. QOL analyses were conducted for all randomized patients (intent-to-treat overall population, ITT-OA) and all patients who had liver metastases at randomization (ITT-LM population). In the ITT-OA population, differences in QWB-SA scores over time between the histamine plus IL-2 group (150 patients) and the IL-2 alone group (151 patients) were not significant (P=0.511, type III F test). In the ITT-LM population (53 histamine plus IL-2 patients and 73 IL-2 alone patients), changes in QWB-SA scores over time favored the histamine plus IL-2 group (P=0.018, type III F-test). In both the ITT-OA and ITT-LM populations, QWB-SA scores deteriorated more quickly over time in the IL-2 alone group than in the histamine plus IL-2 group, resulting in a significant difference in median quality-adjusted survival duration in favor of the histamine plus IL-2 group by 31.3 days in the ITT-OA population (P=0.007, Mann-Whitney U test), and 50.2 days in the ITT-LM population (P=0.011). OSH and GHP scores did not differ between groups. The addition of subcutaneous histamine dihydrochloride to IL-2 treatment improved median quality-adjusted survival duration and did not adversely affect QOL in patients with malignant melanoma. PMID- 12720075 TI - Development and validation of a patient-specific predictive instrument for the need for dose reduction in chemotherapy for breast cancer: a potential decision aid for the use of myeloid growth factors. AB - A predictive instrument for chemotherapy dose reductions would help optimize delivery of planned chemotherapy and rationalize the use of myeloid growth factors. We analyzed data on 833 women with breast cancer treated with cyclophosphamide, doxorubicin, and fluorouracil, for six cycles in two phase III clinical trials. From the first study ( n=323), we generated a logistic regression model that predicts an individual patient's probability of receiving significantly reduced chemotherapy, defined as less than 85% of the planned dose over cycles 2-6, using data generated from cycle 1. The model was validated on data from the second study ( n=510). The predictive model's variables include nadir absolute neutrophil count (ANC) in cycle 1 (OR: 0.14, 95% CI: 0.06-0.30, P<0.001) and percent drop of platelets between day 1 and the nadir in cycle 1 (OR: 1.04, 95% CI: 1.02-1.05, P<0.001). Both variables are dose adjusted based on the chemotherapy cycle 1 dose. The model's discriminatory performance was good (ROC area=0.82), as was the calibration of predicted with actual frequencies of dose reductions. In the validation dataset, model variables remained significant, with an ROC area of 0.78 and good calibration. In summary, we devised and validated a predictive instrument that uses data from a patient's first cycle of chemotherapy to compute the probability of requiring a significant chemotherapy dose reduction on subsequent cycles. This instrument could help clinicians select patients who will benefit from early administration of myeloid growth factors. PMID- 12720076 TI - Non-albicans Candida is the most common cause of candidemia in pediatric cancer patients. AB - GOALS: Candidemia is a serious infection that can severely complicate the care of children with cancer. We sought to determine the spectrum of Candida species in children with cancer, since effective therapy may depend on the species involved. PATIENTS AND METHODS: A retrospective review of candidemia episodes in our pediatric oncology patients over a 9-year period was conducted. During this period azole prophylaxis was not routine in this group. RESULTS: 38 episodes of candidemia were identified: C. albicans 29%, C. tropicalis 26%, C. parapsilosis 24%, C. krusei 8%, C. glabrata 8%, and C. lusitaniae 5%. Non-albicans Candida was common in patients not receiving azole prophylaxis. Species typically susceptible to azoles were common among patients not using azoles. Death attributed to the fungal infection occurred in 21% of episodes, with nearly all the deaths occurring in patients with C. albicans and C. tropicalis. CONCLUSIONS: C. albicans is not the predominant species in pediatric oncology patients experiencing candidemia, even in azole-naive patients. PMID- 12720078 TI - Congenital urinary tract obstruction: Proceedings of the State-Of-The-Art Strategic Planning Workshop-National Institutes of Health, Bethesda, Maryland, USA, 11-12 March 2002. PMID- 12720077 TI - Febrile neutropenia and Fusobacterium bacteremia: clinical experience with 13 cases. AB - OBJECTIVES: To assess the disease spectrum of Fusobacterium bacteremia in our neutropenic patients and review the literature. METHODS: This was a 6.5-year retrospective study in which all the records of neutropenic patients with Fusobacterium bacteremia were analyzed. RESULTS: Fusobacterium bacteremia was found in 13 neutropenic patients, 10 with hematological malignancies and 3 with solid tumors. The standard clinical presentation was that of primary bacteremia with benign evolution under antibiotics with anaerobic coverage. Most patients presented with oral mucositis as the probable portal of entry. Coinfection with other germs was documented in four patients. No patient had a localized infection documented. Most patients were receiving ciprofloxacin chemoprophylaxis. None of the patients had catheter-related infection. All tested strains were susceptible to all standard anaerobic agents. Fusobacterium spp. were responsible for 5% of bacteremias in neutropenic patients in our hospital during the last 6.5 years. CONCLUSION: Fusobacterium bacteremia is a possible cause of febrile neutropenia, especially in the setting of quinolone prophylaxis and oral mucositis after intense chemotherapeutic regimens. We think that its benign outcome if there is no localized infection detected does not justify the use of antianaerobic prophylaxis. Combination of beta-lactams and beta-lactamase inhibitors is a safe and reasonable treatment. PMID- 12720079 TI - Incidence of pediatric IgA nephropathy. AB - Immunoglobulin A (IgA) nephropathy progresses without apparent signs. The only available means of knowing the precise incidence of this disease in children is to analyze data from the school urinary screening system. Students between 6 and 15 years old with IgA nephropathy detected by school urinary screening from 1983 to 1999 in Yonago City, Japan, were examined. In addition, two hospitals with pediatric care units administered a questionnaire and patients with pediatric IgA nephropathy detected by means other than the school urinary screening program were recruited. Of 270,902 children examined by the school urinary screening system, 29 demonstrated suspected chronic nephritis on the basis of abnormal urinalysis and underwent renal biopsy procedures. Among the 29 children, 14 cases of IgA nephropathy were confirmed. During the same period, of the children examined by means other than the school screening program, 20 demonstrated suspected renal disease and underwent renal biopsies. Among these 20 children, 6 cases of IgA nephropathy were diagnosed. In all, 20 cases of pediatric IgA nephropathy were detected over the past 17 years. The incidence rate of pediatric IgA nephropathy was 4.5 cases/year per 100,000 children under 15 years. The incidence of pediatric IgA nephropathy is higher than that of idiopathic nephrotic syndrome. PMID- 12720081 TI - A girl with rickets and nephrocalcinosis. AB - A 5-year-old girl presented with short stature. She was found to have rickets due to renal phosphate wasting and nephrocalcinosis. Serum parathyroid hormone was suppressed, 25-OH vitamin D was within the normal range, and 1,25-(OH)(2 )vitamin D was elevated. In addition, she had hypercalciuria, proteinuria, which was partially tubular in origin, and a reduced glomerular filtration rate of 58 ml/min per 1.73 m(2). Treatment with phosphate supplements resulted in healing of the rickets and normalization of the serum 1,25-(OH)(2 )vitamin D level. This patient is an example of hypercalciuric rickets, most likely due to an inherited disorder of phosphate metabolism. Hypercalciuric rickets can be inherited as an autosomal recessive as well as autosomal dominant trait. PMID- 12720080 TI - Familial hypomagnesemia with hypercalciuria and nephrocalcinosis. AB - Familial hypomagnesemia with hypercalciuria and nephrocalcinosis (FHHNC) is a rare autosomal recessive tubular disorder that is frequently associated with progressive renal failure. The primary defect is related to impaired tubular reabsorption of magnesium (Mg) and calcium (Ca) in the thick ascending limb of Henle's loop. We have studied seven Arab patients with this syndrome who belong to four different families. The mean age at first presentation was 1.5+/-1.3 years (range 0.1-3 years) and at diagnosis 5.9+/-4.3 years (range 0.5-12 years). The presenting features were convulsions and carpopedal spasms (5 patients), polydipsia and polyuria (2 patients), rickets (2 patients), and recurrent urinary tract infections (1 patient). Bilateral nephrocalcinosis was observed in all patients. All patients had hypomagnesemia with a mean serum Mg of 0.45+/-0.09 mmol/l, an inappropriately high urine Mg of 2.07+/-0.73 mmol/24 h or fractional excretion of 15.3+/-7.1%, high urine Ca excretion of 4.1+/-1.2 mmol/24 h or urine Ca to creatinine ratio of 2.6+/-1.6, and normal serum potassium level of 4.4+/ 0.34 mmol/l. All patients received Mg supplements and thiazide but exhibited slow worsening of their kidney function. After a mean follow-up of 4.4+/-3.9 years, one patient progressed to end-stage renal failure (ESRF). In conclusion, we report seven Arab patients with FHHNC syndrome. The clinical and biochemical data were similar to previous reports. However, they tend to show a slower rate of progression to ESRF. PMID- 12720084 TI - Herbivory modifies conifer phenology: induced amelioration by a specialist folivore. AB - Herbivory by Zeiraphera canadensis Mut. & Free. (Lepidoptera: Tortricidae), an early season folivore of white spruce [ Picea glauca(Moench) Voss], has been associated with a shift in the timing of bud burst by its host during the subsequent year. We tested the hypothesis that a herbivory-induced shift in the phenology of bud development improves the window for colonisation of white spruce buds by Z. canadensis. Feeding on cortical tissue of elongating shoots caused the destruction of apical buds and an interruption of apical dominance in the year following herbivory. White spruce compensated for damage with the activation of dormant buds; mainly at proximal positions along shoots. As a result, half of all active buds on previously damaged branches were located immediately adjacent egg sites (i.e. previous year's bud scales), whereas <10% of active buds on intact shoots were situated there. More than 40% of newly emerged larvae colonised the basal buds of damaged shoots versus just 10% for intact shoots. Previous herbivory also influenced the initiation of bud burst. All buds flushed 2 days earlier on damaged shoots and date of bud burst was inversely correlated to bud density, indicating that short damaged shoots with large numbers of buds were stronger sinks for nutrients required for bud development. Egg hatch was best synchronized with early bursting buds on damaged branches. As a consequence, 89% of first-instar larvae successfully colonised buds on damaged branches while only 55% were successful on undamaged branches. Improved survival of larvae in the year following herbivory was a direct result of the evolved response by white spruce to the interruption of apical dominance. The pattern of herbivory by Z. canadensis may have evolved as a strategy to enhance the quality of white spruce for their offspring. PMID- 12720083 TI - Treatment data during pediatric home peritoneal teledialysis. AB - Peritoneal teledialysis (telePD) is a modem-based communication link between the patients' cyclers and a computer in the dialysis unit that allows the transmission and storage of a series of automated peritoneal dialysis (APD) treatment data. In order to evaluate the usefulness of telePD in quantifying the problems that may occur during pediatric APD, we retrospectively studied four patients with a median age of 14.1+/-1.8 years during their initial months of telePD. The selection criteria were potential non-compliance in two cases (patients 1 and 2) and catheter malposition or fibrin occlusion in two (patients 3 and 4). The patients were treated using a Fresenius PD Night Cycler with teledialysis software. Thirty consecutive treatments per patient in the 1st and 4th months were examined, and a series of treatment parameters was calculated. The percentage of treatments with alarms and the number of alarms per treatment were high in both the 1st and the 4th month, particularly in patients 3 and 4. The main causes of the alarms were tube kinking, catheter malfunction, fibrin occlusion, and failure of electrical power. The number of shortened treatments significantly decreased in the 4th month of telePD. One non-compliant family was identified during the 1st month of PD, but psychosocial support helped to decrease the number of shortened treatments due to non-compliance in the 4th month. During the 4th month of telePD, the dwell time/total treatment time ratio (which represents the time of contact between the peritoneum and dialysis fluid) increased as a result of technical interventions aimed at reducing the infusion plus drain time. In conclusion, telePD proved to be useful in detecting and solving the clinical and technical problems of APD. PMID- 12720085 TI - Sympatry and allopatry in two desert ant sister species: how do Cataglyphis bicolor and C. savignyi coexist? AB - Two extremely morphologically similar sister species of desert ants, Cataglyphis bicolor and C. savignyi, exhibit broadly overlapping distributional ranges within Tunisia. In order to analyse the microhabitats of C. bicolor and C. savignyi within the sympatric and allopatric areas of both ant species, the plant species located at 113 different nest sites of the two ant species were determined. In the sympatric area, the two species exhibit a clear-cut nest site segregation. This is not the case in the allopatric areas. Hence the two species differentiate their microhabitat only when they are sympatric. The plant species associated mainly with the nest sites of C. bicolor indicate that this species prefers a type of vegetation that needs irrigation. This is in contrast to the nest sites of C. savignyi, which are usually found around plants that characterize typical dry steppe areas. As the ants' foraging paths recorded in the sympatric area reveal, C. bicolor performs significantly shorter foraging runs with respect to both length and time, and covers a much smaller foraging range than C. savignyi does. This result reflects the fact that the microhabitat occupied by the colonies of C. bicolor is richer in food abundance. When direct interspecific interactions were investigated by placing a bait midway between two heterospecific nests, C. bicolor foragers dominated over those of C. savignyi. The same dominance of C. bicolor over C. savignyi occurred in laboratory experiments. These results suggest that the dominant species drives the subordinate one out of the high quality microhabitats, and that the subordinate species is forced to survive in the less lucrative habitats. In conclusion, coexistence seems to be maintained by the asymmetric competitive relationship between the two species and the fact that the subordinate species has the ability to endure in the less favourable microhabitat. PMID- 12720082 TI - Renal function following hematological stem cell transplantation in childhood. AB - Renal function greatly influences mortality rates in the early phase following hematological stem cell transplantation (HSCT) in childhood, as well as the quality of life in long-term survivors. Nevertheless, the number of studies in pediatric populations is limited and some important aspects of kidney function after HSCT have only been elucidated in adults. The incidence of acute renal failure (ARF) immediately after HSCT in pediatric patients is between 25% and 50%, with 5%-10% of children requiring renal replacement therapy. Doubling of serum creatinine is associated with a twofold increase in mortality. However, the need for dialysis leads to a further increase in mortality rates to 80%-90%. Specific renal syndromes appear at different times following HSCT, revealing a similar pattern in children and adult patients. In both children and adults, impaired renal function associated with liver impairment (hepatorenal syndrome) is the most important cause for ARF. Therapeutic approaches have not been able to reduce the frequency or to improve outcome so far. In adults surviving long term, bone marrow transplant (BMT) nephropathy is the most frequent renal complication, although a considerable variation in incidence (up to 70%) has been published, partly due to various definitions and manifestations. Little is known about the long-term outcome of renal function in patients treated with HSCT in childhood. However, chronic renal failure has been reported in 0%-28%, but no end-stage renal failure has been published so far. Tubular function following HSCT is rarely investigated, although its impact on long-term survivors of BMT in childhood might be of some importance, especially for growth and bone metabolism. PMID- 12720086 TI - Disruption of exon definition produces a dominant-negative growth hormone isoform that causes somatotroph death and IGHD II. AB - Isolated growth hormone deficiency type II (IGHD II) is characterized by short stature due to dominant-negative mutations of the human growth hormone gene (GH1). Most of the known mutations responsible for IGHD II cause aberrant splicing of GH1 transcripts. We have recently shown that mutations that cause exon 3 skipping and produce a dominant-negative 17.5-kDa isoform in humans also cause a dose-dependent disruption of GH secretory vesicles when expressed in GC cells and transgenic mice. We show here that overexpression of the dominant negative 17.5-kDa isoform also destroys the majority of somatotrophs, leading to anterior pituitary hypoplasia in transgenic mice. It is, therefore, important to understand the regulation of GH1 splicing and why its perturbation causes IGHD II. We demonstrate that dual splicing enhancers are required to ensure exon 3 definition to produce full-length 22-kDa hormone. We also show that splicing enhancer mutations that weaken exon 3 recognition produce variable amounts of the 17.5-kDa isoform, a result which could potentially explain the clinical variability observed in IGHD II. Non-canonical splicing mutations that disrupt splicing enhancers, such as those illustrated here, demonstrate the importance of enhancer elements in regulating alternative splicing to prevent human disease. PMID- 12720087 TI - The integration site of the iga gene in commensal Neisseria sp. AB - An IgA1 protease is produced by the human pathogens Neisseria gonorrhoeae and N. meningitidis but not by related non-pathogenic, commensal, Neisseria species. In this study, the chromosomal iga locus was characterized in the N. gonorrhoeae strain MS11 and compared to corresponding loci in N. meningitidis and commensal Neisseria species. In N. gonorrhoeae, the genes trpB and ksgA were found immediately downstream of iga. In addition to comL and comA, a homolog of the Escherichia coli YFII gene was identified upstream of iga. Each gene in the iga region (YFII and comL, comA and iga, and trpB and ksgA) is transcribed in the opposite direction to its neighbors. The comL/ comA and iga/ trpB pairs each have a transcriptional terminator in the correct position for joint use. These terminators contain the common gonococcal DNA uptake sequence (DUS). A highly conserved direct repeat of 25 bp located immediately adjacent to the iga gene in N. gonorrhoeae was also found in N. meningitidis. In Southern hybridization experiments, no homology to iga was detectable in the chromosomal DNAs of the commensal species N. mucosa, N. lactamica, N. flavescens, N. cinerea, N. subflava, N. flava, N. sicca or N. elongata. When N. gonorrhoeae comL and trpB were used as probes, signals were detected on the same restriction fragment in six of the eight species. This indicated that commensal Neisseria species share a possible integration site for the iga gene between comA and trpB. The region between comA and trpB was therefore amplified by PCR. The fragment obtained from N. lactamica showed a high degree of homology to gonococcal comA and trpB, respectively, but iga was replaced by a sequence of 13 bp that shows no homology to any known gonococcal sequence. Our data suggest that iga was acquired by a common ancestor of N. gonorrhoeae and N. meningitidis rather than being distributed by horizontal gene transfer. N. lactamica, which is more closely related to N. gonorrhoeae than other commensals, may have lost iga by deletion. PMID- 12720088 TI - Molecular cloning and immunolocalization of the 17 kDa myoglobin of Clonorchis sinensis. AB - We purified the 17 kDa protein abundant in Clonorchis sinensis crude extracts. The N-terminal amino acid sequence of this protein was determined and an oligonucleotide probe synthesized. Using this probe, the cDNA encoding the protein was cloned and sequenced from the C. sinensis cDNA library. It was found to consist of a total of 150 amino acids and to have 41% conserved homology with the myoglobin of the trematodes Paramphistomum epiclitum and Isoparorchis hypselobagri. The gene product over-expressed in the bacterial system was purified and identified as the same molecule in the adult worms. BALB/c mouse sera raised against the adult 17 kDa protein revealed that this myoglobin was distributed throughout the parenchymal tissues except for the eggs and reproductive organs and that the protein may be involved in the survival of C. sinensis in the oxygen-depleted environment of the host. PMID- 12720089 TI - Ultrastructural localization of phosphoglycerate kinase in adult Clonorchis sinensis. AB - Phosphoglycerate kinase (PGK) is an enzyme that produces one ATP molecule in the glycolytic pathway. Clonorchis sinensis is largely dependent on glycolysis for energy production. We performed immunoelectron microscopy on adult C. sinensis by using mouse immune serum raised against recombinant C. sinensis PGK. A high density of gold particles was found in the microvilli of the intestinal epithelium and in lamellae of the sperm duct. PGK was common in the somatic cells of intra-uterine eggs and in excreted products. It was localized with moderate intensity in muscular fibers of the subtegumental muscle layer, and in the myoepithelia of the intestine and excretory bladder. We suggest that PGK plays an essential role in C. sinensis energy production for movement via muscle contraction. PMID- 12720090 TI - Detection of bacterial contamination in starch and resin-based papermaking chemicals using fluorescence techniques. AB - Rapid fluorescence techniques were evaluated for the detection of bacterial contaminants in papermaking chemicals including starch and the resin-based sizes and starch slurries used in the paper industry. Viable and non-viable bacterial cells were visualised by fluorescent probes and detected by epifluorescence microscopy and flow cytometry. The best discrimination ability was obtained with the fluorescent probes LIVE/DEAD and SYBR Green, based on the staining of cellular nucleic acid, and ChemChrome V3, which demonstrated cellular enzymatic activity. The process samples had to be diluted and filtered before fluorescence staining and analysis because they were viscous and contained solid particles. Fluorescence microscopic counts of bacteria in highly contaminated process samples were similar to plate counts, but flow cytometric enumeration of bacterial cells in process samples yielded 2- to 10-fold lower counts compared with plate counts, depending on the consistency of the sample. The detection limits in flow cytometric analysis and in epifluorescence microscopy were 10(3) 10(6) cells ml(-1) and 10(5)-10(6) cells ml(-1), respectively. Intrinsic bacterial contamination was detectable with fluorescence techniques and highly contaminated process samples could be analysed with fluorescence methods. PMID- 12720091 TI - Familiality in simple and complex disease. AB - Establishing the existence of a genetic contribution to disease is the first step in identifying the underlying gene defect(s). This can present a considerable challenge, particularly in diseases where the genetic contribution is complex in nature. Clinically well-characterized cohorts and large inter-institutional collaborations will be required to identify genes involved in complex disease. PMID- 12720092 TI - Cutaneous sympathetic function in patients with multiple system atrophy. AB - Some procedures increase the sweat output (SSwR; sympathetic sweat response) and reduce the cutaneous blood flow (SVR; skin vasomotor reflex) in the hand. We evaluated SSwRs and SVRs to deep inspiration, mental arithmetic, exercise, and tactile stimulation in 40 MSA patients and 15 healthy controls. We also conducted head-up tilt tests and R-R interval variation tests (CV(R-R)). SSwRs were present in all controls, but absent in 19 (47.5 %) of the MSA patients. The mean SSwR amplitudes in the MSA group were significantly lower than those in the control group. SVRs were evoked in all subjects except 3 MSA patients. There were no marked differences in SVR amplitudes between the two groups. Orthostatic hypotension and low CV(R-R) values were seen in 18 (45 %) and 13 (32.5 %) of the MSA patients, respectively. SSwR amplitudes correlated significantly with postural fall in blood pressure and CV(R-R) values in the MSA group. SSwRs were absent in about half of the MSA patients, and the SSwR results correlated with those of the cardiovascular autonomic tests. The SVRs were not severely disturbed in the MSA patients. We considered SSwR a useful index for the detection of autonomic dysfunction in MSA. PMID- 12720093 TI - Primary hyperhidrosis--evidence for autosomal dominant inheritance. AB - Primary hyperhidrosis is a neurogenic disorder of unknown cause characterized by excessive sweating in the palmar surface of the hands, armpits, groin and feet. In the course of a therapeutic trial for primary hyperhidrosis, 62 % of patients reported a positive family history. Examination of these pedigrees demonstrated a sibling recurrence risk of lambdas = 29-48 and an offspring recurrence risk of lambdao = 41-68 indicating that hyperhidrosis can be an inherited condition. The pattern of inheritance suggests an autosomal dominant mode of transmission with incomplete disease penetrance. PMID- 12720094 TI - Agreement of two different methods for measurement of heart rate variability. AB - BACKGROUND: The widespread use of affordable devices with sufficient precision for measurement of heart rate variability (HRV) might lead to early detection of abnormalities in a large number of high-risk patients and athletes. The purpose of this study was to determine the limits of agreement of two devices for measuring HRV parameters differing in price and assumed precision. SUBJECTS AND METHODS: 36 healthy subjects (22 men and 14 women) with a mean age of 27.4 (SD 11.1) years were included. The two devices used for comparison were PowerLab with Chart software as the reference golden standard, and Polar Transmitter/Advantage with Precision Performance software, respectively. Measurements included the following heart rate variability parameters: heart rate, range of R-R-interval duration, SDNN, rMSSD, total Power, VLF power, LF power, and HF power. Measurements were taken during metronomic respiration over a total period of 3 minutes. Statistical analysis was performed according to Bland and Altman and by means of scatterplots and Spearman correlation coefficients. RESULTS: Good agreement was found for heart rate (95 % CI of limits of agreement: -0.7-0.6 bpm; r = 0.999), range of duration of R-R-intervals (95 % CI: -18.9-17.0 ms; r = 0.997), rMSSD (95 % CI: -1.5-2.5 ms; r = 0.999), and SDNN (95 % CI: -3.0-3.1 ms; r = 0.997). Correlation of measurements was high for the variables total Power, VLF power, LF power, and HF power. Analysis of method agreement for frequency domain variables was statistically not feasible. CONCLUSION: The level of agreement for the analyzed time domain variables between the reference golden standard and the inexpensive device is sufficient to permit initial screening by family doctors, and self-administration by high-risk patients and athletes. PMID- 12720095 TI - Water drinking as a potential treatment for idiopathic exercise-related syncope: a case report. AB - We report a 20-year-old sportsman with frequent attacks of lightheadedness, chest pain, blurred vision and falls during and shortly after exercise. Cardiac and pulmonal evaluation and routine autonomic function tests were normal apart from a relatively high resting heart rate (70 bpm) compared to endurance-trained men. In view of the relation to exercise, the patient was asked to cycle with maximal effort on an ergometer with continuous blood pressure (BP), heart rate (HR) and electroencephalogram (EEG) registration. Immediately after cessation of exercise a brief hypotensive period (75/45 mmHg) occurred together with sinus tachycardia (180 bpm) during which the patient experienced his typical complaints. We hypothesized that our patient's symptoms were primary related to sympathetic failure. As water drinking has been demonstrated to raise sympathetic activity rapidly, we undertook a second cycling test after ingestion of 1000 mL tap water. Symptoms nor hypotension recurred. Because of the short lasting pressor effect and its minimal side effects, we suggest water drinking as simple and possible effective therapy for idiopathic exercise-related syncope. PMID- 12720096 TI - The effect of intranasal desmopressin on nocturnal waking in urination in multiple system atrophy patients with nocturnal polyuria. AB - Nocturnal waking in urination (nocturia) is a common feature in patients with multiple system atrophy (MSA). Degeneration of arginine vasopressin (AVP) neurons in the suprachiasmatic nucleus in this disorder may lead to nocturnal polyuria, which has been treated with intramuscular desmopressin. We also prescribed 5 microg of intranasal desmopressin once a night in 3 MSA patients, who had nocturnal polyuria with abnormal circadian rhythm of the plasma AVP. During the treatment, all patients showed an improvement in their nocturia without serious adverse effects. PMID- 12720097 TI - Clinical and pathological study of gastric cancer with ovarian metastasis. AB - BACKGROUND: The aim of this study was to determine the treatment strategy for ovarian metastases from gastric cancer, by a retrospective study of the treatment results. METHODS: We reviewed the records of patients with ovarian metastases from primary gastric cancer. Ovarian metastases were found in 24 of 897 female patients with gastric cancer. Of these, 21 patients with histologically proven disease were studied. RESULTS: Ovarian metastasis was detected before the primary gastric cancer in 1 patient, simultaneously in 6, and after in 14. Ovarian tumors were detected by computed tomography (CT) in a majority of patients (95%), while uterine tumors were detected in only 29%. Metastasis to the uterus was histologically examined in 14 tumors and confirmed in 11 tumors. All patients with positive endometrial cytology had uterine metastases. Total abdominal hysterectomy was performed with bilateral salpingo-oophorectomy in 12 patients and with unilateral resection in 2. In these 14 patients, 5 were curatively operated. In the clinical course, all patients developed multiple metastases, and patients suffered peritoneal dissemination. None survived for longer than 3 years. The median survival time after ovarian metastases (MST) was 10.3 months for all patients; 3.6 months in patients in their sixties, and 12.5 months in those in their fifties. Survival was significantly longer in patients who underwent curative resection (MST, 30.4 months) compared with those who had noncurative resection (MST, 10.3 months). CONCLUSION: The prognosis for ovarian metastasis of gastric cancer was poor without curative resection. Because of frequent microscopic metastases to the uterus, total hysterectomy with bilateral oophorectomy is recommended if curative resection is possible. PMID- 12720098 TI - Relationships between the expression of thymidylate synthase, dihydropyrimidine dehydrogenase, and orotate phosphoribosyltransferase and cell proliferative activity and 5-fluorouracil sensitivity in colorectal carcinoma. AB - BACKGROUND: The site of action of the 5-fluorouracil (5-FU) antitumor effect has been explicated in recent years. Many studies have investigated enzymes involved in 5-FU metabolism in attempts to predict this effect, and a correlation of enzyme activity with the 5-FU drug sensitivity test has been reported. The aim of this study was to identify the biochemical response determinants of 5-FU. Additionally, we aimed to clarify the association between cell proliferative activity and the response to 5-FU of colorectal cancer. METHODS: Our research subjects were 54 patients with colorectal carcinoma who had undergone operations between August 1999 and July 2001 in our department. Assays of thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD), and orotate phosphoribosyltransferase (OPRT) activities in colorectal carcinoma tissue and assays of 5-FU sensitivity by the collagen gel droplet embedded culture drug sensitivity test (CD-DST) were conducted to investigate the relationships between each enzyme activity and 5-FU sensitivity. In addition, the proliferative activity of cancer cells was evaluated with Ki-67 antibody, and the relationship of this activity to each enzyme activity and 5-FU sensitivity were investigated. RESULTS: 5-FU sensitivity was high in the low-TS-activity group and in the high OPRT-activity group. Cancers with high cell proliferative activity showed good sensitivity to 5-FU, and TS and OPRT activities were high in such cancers. CONCLUSION: The results suggest that OPRT activity can predict sensitivity to 5 FU, and high OPRT activity may cause good 5-FU sensitivity in cancers with high cell proliferative activity. PMID- 12720099 TI - Elevated serum epidermal growth factor receptor level is correlated with lymph node metastasis in lung cancer. AB - BACKGROUND: Using an enzyme immunoassay for epidermal growth factor receptor (EGFR), we investigated whether serum EGFR levels could be used as predictors of the development and extent of lung cancer. METHODS: The study included 106 lung cancer patients and 16 patients with nonmalignant thoracic disease. Serum samples were collected before clinical treatment. RESULTS: There was no difference between serum EGFR levels in patients with lung cancer (21.275 +/- 22.035 fm/ml) in comparison with those in nonmalignant-disease controls (22.630 +/- 7.330 fm/ml; P = 0.8083). However, lung cancer patients with lymph node metastasis (23.515 +/- 20.065 fm/ml) had significantly higher EGFR levels compared with those in patients without lymph node metastasis (16.390 +/- 10.970 fm/ml; P = 0.0228). The serum EGFR levels were similar in samples from lung cancer patients with various pathological subtypes. There was no difference in the prognosis between the lung cancer group with normal EGFR levels (<850 ng/ml) and the group with elevated EGFR levels (>850 ng/ml). CONCLUSION: Serum EGFR levels may serve as a marker that can be used as an indicator of lymph node metastasis in lung cancer. However, there was no difference between levels in patients with lung cancer and those in nonmalignant-disease controls, indicating that the measurement of serum EGFR levels was of limited value in the detection of lung cancer. PMID- 12720100 TI - Paradoxically abundant expression of Bcl-2 and adrenomedullin in invasive cervical squamous carcinoma. AB - BACKGROUND: The purpose of this study was to identify the role of Bcl-2 protein and adrenomedullin (AM) expression in relation to apoptosis in the neoplastic changes of squamous epithelium of the uterine cervix. METHODS: Apoptosis in sections of normal cervical epithelium, cervical intraepithelial neoplasia (CIN), and invasive squamous cell carcinoma was determined by terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate nick-end labeling (TUNEL) assay. Bcl-2 protein and AM expression were analyzed using the avidin-biotin immunoperoxidase method. RESULTS: On the basis of the TUNEL assay, the apoptosis positive rate of the nuclei in early invasive squamous carcinoma and in bulky invasive squamous carcinoma was significantly higher than that in normal cervical epithelium, but no such increase in the apoptosis-positive rate was noted in the nuclei in CIN. Immunohistochemical staining revealed that Bcl-2 protein and AM were immunolocalized in the cytoplasm of invasive squamous carcinoma cells, but not in either CIN lesions or normal cervical epithelium. Bcl-2 protein expression was more prominent in bulky invasive squamous carcinoma cells than in early invasive squamous carcinoma cells. AM expression was also more abundant in bulky invasive squamous carcinoma compared with that in early invasive squamous carcinoma. CONCLUSION: The present results suggest that the expression of Bcl-2 protein and AM in invasive squamous carcinoma may play crucial roles in selecting carcinoma cells resistant to apoptosis and in promoting malignant progression. PMID- 12720101 TI - Cytoreductive surgery combined with organ resection for advanced ovarian carcinoma. AB - BACKGROUND: The survival effects of combined organ resection in cytoreductive surgery for advanced ovarian carcinoma with regard to the site and the number of organs involved have not yet been clarified. METHODS: Data obtained from 143 patients with stage III/IV ovarian carcinoma were used for analysis. Combined organ resection (COR) was employed in 21 patients in whom optimal cytoreduction (defined as a residuum chloride>fluoride. Full optimization was performed at the ground and transition states in the molecule at both MP2 and B3LYP levels. The barrier to internal rotation around the C-N single bond in the three molecules was calculated to be about 4-5 kcal mol(-1). The vibrational frequencies were computed at the DFT-B3LYP level and the calculated infrared and Raman spectra of the cis- trans mixture of the three molecules were plotted. Complete vibrational assignments were made on the basis of normal coordinate calculations for both stable conformers of the three molecules. PMID- 12720115 TI - The effect of postsurgical administration of a selective cyclo-oxygenase-2 inhibitor on the healing of intrabony defects following treatment with enamel matrix proteins. AB - Regenerative treatment with enamel matrix proteins (EMD) has been shown to promote regeneration in intrabony periodontal defects. However, up to now various postoperative regimens such as the routine administration of nonsteroidal anti inflammatory drugs (NSAIDs) were often used in combination with enamel matrix proteins. Therefore, it cannot be excluded that the results might have been influenced by the effect of the postoperative medication. The aim of this randomized, controlled, blinded, clinical investigation was to determine the effect of postsurgical administration of a selective cyclo-oxygenase-2 inhibitor on the healing of intrabony periodontal defects following regenerative periodontal surgery with EMD. Twenty two patients, each of whom exhibited one deep intrabony defect, were randomly treated with either EMD plus a selective cyclo-oxygenase-2 (COX-2) inhibitor (test) or with EMD alone (control). The postoperative regimen consisted of oral administration of 12.5 mg rofecoxib twice daily for 14 days. The following parameters were recorded at baseline and at 6 months by the same calibrated and blinded investigator: plaque index (Pl), gingival index (GI), bleeding on probing (BOP), pocket depth (PD), gingival recession (GR), and clinical attachment level (CAL). Power analysis to determine superiority of the anti-inflammatory treatment showed that the available sample size would yield 70% power to detect a 1 mm difference. No statistical significant differences in any of the investigated parameters between the two groups were observed at baseline. The results show that, in the test group, mean PD decreased from 8.7+/-1.4 mm to 4.7+/-2.0 mm (P<0.001) and mean CAL from 9.7+/ 2.0 mm to 6.5+/-2.1 mm (P<0.001). In the control group, mean PD decreased from 8.6+/-1.6 mm to 4.7+/-1.8 mm (P<0.001) and mean CAL from 9.5+/-1.6 mm to 6.5+/ 2.2 mm (P<0.001). There were no significant differences between the two groups in any of the investigated parameters. Within the limits of the present study, it can be concluded that the systemic administration of a selective COX-2 inhibitor following regenerative periodontal surgery with EMD did not result in additional clinical improvements when compared to treatment with EMD alone. PMID- 12720116 TI - Metal content of biopsies adjacent to dental cast alloys. AB - Single case reports indicate that components of dental alloys accumulate in the adjacent soft tissue of the oral cavity. However, data on a wider range of dental alloys and patient groups are scarce. Therefore, the aim of the present study was to examine the metal content of oral tissues adjacent to dental alloys showing persisting signs of inflammation or other discoloration (affected sites) and of healthy control sites with no adjacent metal restoration in 28 patients. The composition of the adjacent alloys was analyzed and compared to the alloy components in the affected sites. Tissue analysis was performed using atomic absorption spectroscopy. Alloy analysis was performed with energy-dispersive X ray analysis. In the affected sites, the metals Ag, Au, Cu, and Pd prevailed compared to control sites, reflecting the frequency distribution of single metals in the adjacent alloys. In most cases (84%), at least one of the analyzed metals was a component of the alloy and also detected in the tissue. Metal components from almost all dental cast alloys can be detected in adjacent tissue. PMID- 12720117 TI - Defining Waldenstrom's macroglobulinemia. PMID- 12720118 TI - Clinicopathological definition of Waldenstrom's macroglobulinemia: consensus panel recommendations from the Second International Workshop on Waldenstrom's Macroglobulinemia. AB - This presentation represents consensus recommendations for the clinicopathological definition of Waldenstrom's macroglobulinemia (WM), which were prepared in conjunction with the Second International Workshop held in Athens, Greece during September 2002. WM is an uncommon lymphoproliferative disorder characterized primarily by bone marrow infiltration and IgM monoclonal gammopathy. It should be considered a distinct clinicopathological entity rather than a clinical syndrome secondary to IgM secretion. The underlying pathological diagnosis in WM is lymphoplasmacytic lymphoma as defined by the World Health Organization (WHO) and Revised European-American Lymphoma (REAL) classification criteria. The concentration of monoclonal IgM can vary widely in WM and it is not possible to define a concentration that reliably distinguishes WM from monoclonal gammopathy of undetermined significance (MGUS) and other lymphoproliferative disorders. A diagnosis of WM can therefore be made irrespective of IgM concentration if there is evidence on a bone marrow trephine biopsy of bone marrow infiltration by lymphoplasmacytic lymphoma with predominantly an intertrabecular pattern, supported by appropriate immunophenotypic studies. Simple criteria to distinguish patients with symptomatic WM who require therapy from those with asymptomatic WM and MGUS were also proposed. Patients with clinical features attributable to IgM monoclonal gammopathy but no overt evidence of lymphoma are considered to constitute a distinct clinical group and the term "IgM-related disorders" is proposed. PMID- 12720119 TI - Prognostic markers and criteria to initiate therapy in Waldenstrom's macroglobulinemia: consensus panel recommendations from the Second International Workshop on Waldenstrom's Macroglobulinemia. AB - This presentation represents consensus recommendations on prognostic markers and criteria to initiate therapy in patients with Waldenstrom's macroglobulinemia (WM), which were prepared in conjunction with the Second International Workshop held in Athens, Greece during September 2002. The panel recommended that initiation of therapy should not be based on the IgM level per se since this may not correlate with the clinical manifestations of WM. The consensus panel agreed that initiation of therapy was appropriate for patients with constitutional symptoms such as recurrent fever, night sweats, fatigue due to anemia, or weight loss. The presence of progressive, symptomatic lymphadenopathy or splenomegaly provide additional reasons to begin therapy. The presence of anemia with a hemoglobin value of 4 x 10(9)/L), bone marrow lymphoplasmacytoid infiltration (>10%), erythrocyte sedimentation rate (>40 mm/h), and detectable Bence Jones proteinuria were significantly related with evolution probability. At multivariate analysis, paraprotein level (P <.0001), hemoglobin level (P <.05), and lymphocytosis (P <.0001) independently predicted malignant evolution (P <.0001). In conclusion, patients with asymptomatic IgM-MG showing hematological features predictive of progression should be carefully monitored in view of an early treatment of the disease. PMID- 12720132 TI - Malignant transformation in IgM monoclonal gammopathy of undetermined significance. AB - Monoclonal gammopathy of undetermined significance (MGUS) is a frequent disorder characterized by the presence of a small serum M-protein in individuals with no evidence of multiple myeloma (MM), Waldenstrom's macroglobulinemia (WM), or primary amyloidosis (AL). Although one fourth of these individuals will develop a malignant disease, there are no well-established predictors of outcome, particularly in the IgM type MGUS. Among 434 patients diagnosed with MGUS from 1970 to 2001 with a minimum follow-up of 1 year, 52 (27 men and 25 women; median age, 67 years) of IgM type were identified. After a median follow-up of 5 years, five patients (9.6%) have developed WM. The risk of transformation was 13.3% (95% confidence interval [CI], 0 to 27) and 27.7% (95% CI, 0.3 to 55.1) at 10 and 20 years, respectively. The variables significantly associated with transformation were the proportion of bone marrow plasma cells (BMPC) and the percentage of bone marrow lymphocytes (BML). No significant differences in the risk of transformation were found between IgM MGUS and the remaining MGUS types. Thus, in IgM MGUS the rate of transformation was similar to the risk observed in other MGUS types, the percentage of BMPC and BML being the features significantly associated with evolution into WM. PMID- 12720133 TI - Despite apparent morphologic and immunophenotypic heterogeneity, Waldenstrom's macroglobulinemia is consistently composed of cells along a morphologic continuum of small lymphocytes, plasmacytoid lymphocytes, and plasma cells. AB - We studied the clinical, morphologic, and immunophenotypic features of 26 Waldenstrom's macroglobulinemia (WM) cases, each with an IgM spike of >or=3.0 g/dL. The neoplastic cells were consistently composed of a spectrum of small lymphocytes, plasmacytoid lymphocytes, and plasma cells. Bone marrow (BM) involvement ranged from 10% to 90%, showed four histologic patterns (nodular [75%], interstitial [75%], paratrabecular [42%], and diffuse [4%]), two histologic subtypes (lymphoplasmacytic [87%] and lymphoplasmacytoid [13%]), and several cytologic variants (monocytoid [n = 2], signet-ring cell [n = 2], and hairy cell leukemia-like [n = 1]). By flow cytometry (FC), all cases expressed monoclonal surface immunoglobulin, CD19, and CD20. Most cases (58%) lacked expression of CD5, CD10, and CD23. However, variants such as CD5(+)CD10(-)CD23(-) (n = 3), CD5(+)CD10(-)CD23(+) (n = 1), and CD5(-)CD10(+)CD23(+/-) (n = 2) were seen. At last follow-up, 18 of 26 patients were alive (median survival, 94 months). Causes of death included WM (n = 1), large cell lymphoma (n = 1), acute myeloid leukemia (likely therapy-related [n =2]), and nonhematologic/unknown. When individual WM cases are compared, apparent morphologic diversity is suggested. However, every WM case is comprised of cells along a morphologic continuum from small lymphocytes to plasma cells, delineating a uniform, consistent pathology. As WM shows immunophenotypic heterogeneity, morphology must be the cornerstone of the diagnosis. PMID- 12720135 TI - Developing diagnostic criteria in Waldenstrom's macroglobulinemia. AB - Waldenstrom's macroglobulinemia (WM) is a poorly characterized B-cell lymphoproliferative disorder. There are a relatively limited number of detailed clinicopathological assessments, while the majority of clinical trials have been nonrandomized, single-institution phase II studies. Unfortunately progress in this disorder has been hindered by a lack of universally accepted diagnostic criteria. It is clear that criteria incorporating clinical, morphological, immunophenotypic, and, ultimately, genotypic parameters are needed for future clinical trails. Following a detailed clinicopathological assessment of 111 patients and a review of the published literature the following diagnostic criteria were proposed for WM: IgM monoclonal gammopathy of any concentration, bone marrow infiltration by small lymphocytes, plasmacytoid cells and plasma cells in a diffuse, interstitial or nodular pattern, and a surface Ig(+)CD19(+)CD20(+)CD5(-)CD10(-)CD23(-) immunophenotype. PMID- 12720134 TI - Immunophenotypic analysis of Waldenstrom's macroglobulinemia. AB - Immunophenotyping has become an essential tool for diagnosis of hematological malignancies. By contrast, for diagnosis of Waldenstrom's macroglobulinemia (WM) immunophenotyping is used only occasionally. From 150 patients with a IgM monoclonal gammopathy we have selected 60 cases with (1) morphological lymphoplasmocytoid bone marrow (BM) infiltration (>20%); (2) IgM paraprotein (>10g/L); and (3) absence of features of other lymphoma types. Immunophenotypic analysis was based on the use of the triple or quadruple monoclonal antibody (MoAb) combinations. To increase the sensitivity of the analysis of antigen expression, selected CD19(+)CD20(+) B cells were targeted. We have also explored the antigenic characteristics of both the plasma cell (PC) and mast cell (MC) compartments present in the BM from 15 WM patients. Clonal WM lymphocytes were characterized by the constant expression of pan-B markers (CD19, CD20, CD22, CD24) together with sIg, predominantly kappa (5:1, kappa:lambda ratio). A high proportion of cases (75%) were positive for FMC7 and CD25, but in contrast to hairy cell leukemia (HCL), these lymphocytes were always negative for CD103 and CD11c. CD10 antigen was also absent in all WM patients and less than one fifth of patients were positive for CD5 and CD23, while CD27, CD45RA, and BCL-2 were present in most malignant cells. In two cases, the coexistence of two different clones of B lymphocytes was identified, and in eight additional cases, intraclonal phenotypic heterogeneity was observed. As far as PCs are concerned, in most patients (85%) the number of PCs was within the normal range (median, 0.36%). The antigenic profile of these PCs differed from that observed in normal and myelomatous PC (CD38(++)CD19(++/-)CD56(-)CD45(++)CD20(+)). In three cases, PCs showed aberrant expression for CD5, CD22, or FMC7. Finally, the number of mast cells was significantly higher (0.058 +/- 0.13) as compared to normal BM (0.019 +/- 0.02) (P <.01), although they were immunophenotypically normal (CD117(+)CD2(-)CD25(-)). PMID- 12720136 TI - Differential diagnosis of Waldenstrom's macroglobulinemia from other low-grade B cell lymphoproliferative disorders. AB - Waldenstrom's macroglobulinemia (WM) is a rare lymphoproliferative disorder (LPD) characterized by lymphoplasmacytic infiltration of the bone marrow (BM) and/or occasionally other tissues and by the presence of a serum monoclonal IgM. The disease belongs to the lymphoplasmacytic lymphoma (LPL) subtype. Whether WM is indeed a separate entity or is merely an IgM-secreting subtype of low-grade B cell lymphoma is still controversial. In our series of 67 patients, WM has a long median overall survival of 110 months, and the male/female ratio is 1.2/1. Clinical features include a wide spectrum of manifestations, many of which may be common to other LPDs. Differential diagnosis is based on: (1) clinical and laboratory features (anemia, organomegaly, lymphadenopathy, IgM paraproteinemia), (2) cell morphology (lymphocytes, lymphoplasmacytes, few plasma cells), (3) histopathology of bone marrow or lymph node, (4) immunophenotype (CD5 expression and the intensity of CD5, CD20, and CD79b antigens may help in discrimination from other LPDs and atypical CLL), and (5) characteristic genetic features (present in other LPDs). Based on the former, diagnosis is usually easy. It may be harder in LPL cases not secreting IgM. We consider that WM should be, for the time being, handled as a separate entity. Further studies are necessary. PMID- 12720137 TI - Asymptomatic Waldenstrom's macroglobulinemia. AB - A study was undertaken to evaluate the frequency and natural history of disease in patients with asymptomatic Waldenstrom's macroglobulinemia (WM). Among 132 consecutive, newly diagnosed patients with monoclonal IgM, 82 (27%) had symptomatic WM indicated by anemia, lymphadenopathy, or splenomegaly. Thirty-one patients had similar clinical features but were asymptomatic and followed without therapy until disease progression. There were 19 patients with monoclonal gammopathy of undetermined significance of IgM type (MGUS). In comparison to overt WM, patients with asymptomatic WM had significantly higher hemoglobin (Hgb) level (median, 12.1 v 9.7 g/dL), lower serum beta(2)-microglobulin (B(2)M) level (median, 2.4 v 3.4 mg/L), and similar IgM peaks (median, 2.2 and 1.8 g/dL). The IgM component was 3.6 g/dL or less in all patients with asymptomatic disease. For asymptomatic WM, median time to disease progression was 6.9 years with rare morbidity. Prognostic factors for early progression were Hgb <11.5 g/dL, B(2)M >or= 3.0 mg/L, and IgM peak >3.0 g/dL. Combinations of these variables defined three risk groups for progression with markedly different median times to progression of >5 years, 2 years, and 0.5 year, respectively. Response rate and survival after institution of treatment were similar to those of patients treated promptly for overt disease. We conclude that, among patients with WM, 27% were asymptomatic with slow disease progression before the need for chemotherapy. Since disease outcomes after treatment were similar to those of patients treated at diagnosis, patients with asymptomatic disease should be identified and followed without treatment for as long as risks of complications remain low. PMID- 12720138 TI - Prognostic factors in symptomatic Waldenstrom's macroglobulinemia. AB - We analyze the prognostic value of the presenting features of a series of patients with symptomatic Waldenstrom's macroglobulinemia who were homogeneously treated. A total of 215 patients (119 males) with a median age of 62.6 years (range, 24.9 to 91.6) were retrospectively analyzed. The median overall follow-up was 57.6 months (range, 0.6 to 281): 58 (0.9 to 281) for living patients and 52.2 (0.6 to 261.3) for those who died. All patients were treated with alkylating agent-based chemotherapy. The overall median survival was 77.2 months, without significant differences based on the duration of the previous monoclonal gammopathy of undetermined significance (MGUS) phase. The multivariate Cox analysis performed on the whole population showed that age, hemoglobin level, and serum albumin level predicted survival. The addition of beta(2)-microglobulin, available in the subgroup of 60 patients diagnosed after 1990, in a Cox stepwise selection showed that this parameter was by far the main prognostic determinant. Application of the Dhodapkar, Morel, and Gobbi scoring systems to this population of patients showed that all three stratified the population into groups with significantly distinct prognoses. A prognostic index based on age, hemoglobin, and albumin is capable of identifying various groups of patients with different therapeutic needs. PMID- 12720139 TI - Prognostic factors in Waldenstrom's macroglobulinemia: description of the complications during the evolution-preliminary results on 101 patients. AB - Data on clinical features observed in patients with Waldenstrom's macroglobulinemia (WM) during follow-up remain limited. Therefore, we evaluated 860 follow-up procedures in 101 patients. Median age was 66 years and 5-year overall survival 72%, with a median follow-up of 36 months in surviving patients. Sixteen patients presented at diagnosis with two or three cytopenias lasting for at least 3 months (multiple cytopenias [MC]), and MC improved after treatment in eight patients, 4 to 18 months later. MC was observed during at least 6 consecutive months in 23 other patients, 2 to 73 months (median, 24) after diagnosis. MC occurred off-therapy in 12 patients, and on-therapy in 11. Regression occurred in three of the former patients, and in seven of the latter (6 to 24 months after completion of treatment; median, 7). Finally, the 4-year estimated cause-specific cumulative incidence was 40% in the 101 patients. A second malignancy was observed in 11 patients, histological transformation in three, and rapid rise of M-component in only six patients. In conclusion, the present analysis pointed out a high incidence of long lasting MC during the evolution of WM, and a low frequency of rapid rise of M component. PMID- 12720140 TI - Prognostic factors and response to fludarabine therapy in Waldenstrom's macroglobulinemia: an update of a US intergroup trial (SW0G S9003). AB - We report an update on a prospective observational trial for Waldenstrom's macroglobulinemia (WM) that called for re-registration to treatment with fludarabine (30 mg/m(2)) upon the development of symptomatic or progressive disease. Patients who did not require therapy for more than 1 year (n = 54) could be distinguished from the 118 untreated patients requiring immediate therapy on the basis of hemoglobin, serum beta(2)-microglobulin (beta2m), C-reactive protein (CRP), albumin, and IgM levels, and lower incidence of extramedullary infiltration. Overall response rate (>or= partial response [PR]) to fludarabine was 38%, with 2% complete remissions (CRs). Event-free and overall survivals were significantly longer in the presence of lower levels of serum beta2m in all cohorts. Using time-dependent covariates, neither the occurrence of response (>or= PR) nor the time to response was associated with superior overall or event free survival. These data support serum beta2m as the dominant prognostic indicator in WM, and show that this factor alone can provide valuable disease risk assessment. Response to therapy using current criteria is not a reliable predictor for survival in this disease. PMID- 12720141 TI - Patterns of clinical presentation, treatment, and outcome in patients with Waldenstrom's macroglobulinemia: a two-institution study. AB - Waldenstrom's macroglobulinemia (WM) is in the World Health Organization (WHO) classification considered to be a clinical syndrome rather than a specific pathologic diagnosis. The clinical manifestations associated with WM relate to direct tumor infiltration, hyperviscosity, and deposition of IgM in various tissues. The indications for and choice of treatment vary considerably and no generally accepted prognostic models exist. The clinical features, treatment, and prognosis of 72 patients with WM seen at one British (n = 36) and one Swedish (n = 36) academic center were therefore compared. Significantly more patients presented with a low albumin concentration (< v > 40 g/L, P <.001), anemia (hemoglobin < v > 120 g/L; P <.001), thrombocytopenia (< v > 150 x 10(9)/L; P <.05), hepatomegaly (P <.001), splenomegaly (P <.01), and lymphadenopathy (P <.01), at St Bartholomew's Hospital (SBH) in comparison to the Karolinska Hospital (KH). Fifty-six percent of SBH patients received chemotherapy immediately following diagnosis as compared to 14% at KH. The median overall survival of all patients was 6.3 years; 4.2 years and 11.0 years at SBH and KH, respectively (P <.001). In univariate analysis, anemia (hemoglobin < 120 g/L) and albumin < 35 g/L (but not <40 g/L) at diagnosis predicted a worse overall survival. The presence of hepatomegaly and/or splenomegaly and/or lymphadenopathy was significantly associated with anemia (P <.001) and hypoalbuminaemia (P <.001). The mean Morel score (including age, albumin, and cytopenias) of patients treated at SBH (2.6) was significantly higher than that of KH patients (1.6; P <.001). These findings illustrate the clinical heterogeneity of WM, most probably explained by differences in referral patterns, and in addition, indicate the need for establishing standard criteria for diagnosis, response to treatment, and prognostic features. PMID- 12720142 TI - Smouldering Waldenstrom's macroglobulinemia: factors predicting evolution to symptomatic disease. AB - Factors predicting evolution to symptomatic disease were investigated in 27 patients with smouldering Waldenstrom's macroglobulinemia (SWM) among 172 patients with Waldenstom's macroglobulinemia (WM), selected on the basis of the following criteria: (1) IgM paraprotein > 3 g/dL, and/or (2) bone marrow (BM) lymphoplasmacytoid (LPC) infiltration >or= 30%, and/or (3) diffuse infiltration pattern on BM biopsy, and (4) no treatment requirement for at least 12 months. Cumulative probability of survival was calculated by means of Kaplan-Meier. The Mantel and Haenszel test and multivariate Cox model were used to identify possible predictors for evolution. At a median follow-up of 79 months (range, 14 to 204), 11 patients (40.7%) showed progression to symptomatic disease, with the median interval from diagnosis being 46 months (range, 12 to 154). Event-free survival (EFS) at 5 and 10 years was 65% (95% confidence interval [CI], 45% to 85%) and 53% (95% CI, 31% to 75%), respectively. At multivariate analysis, paraprotein > 3 g/dL (hazard ratio [HR], 15.1; 95% CI, 3.01 to 75.64; P <.0009) and hemoglobin 3 g/dL and hemoglobin levels or=50% of bone marrow tumor cells (CD19(+), kappa/lambda light chain restricted), respectively, from patients as follows: CD20 (98.3%), CD22 (88.3%), CD40 (83.3%), CD52 (77.4%), IgM (83.3%), MUC1 core protein (57.8%), and 1D10 (50%). Both interpatient and intrapatient tumor clone antigen expression was heterogeneous. Combined mAb therapy might be a useful approach to cope with this variation, and could be tailored to target all members of the tumor clone for an individual patient. PMID- 12720147 TI - CD22 as a target of passive immunotherapy. AB - CD22 is a 135-kd B-cell restricted sialoglycoprotein present in the cytoplasm of virtually all B-lineage cells but expressed on the B-cell surface only at mature stages of differentiation. In humans, the vast majority of IgM(+)IgD(+) B cells express cell-surface CD22, while in lymphoid tissues CD22 expression is high in follicular mantle and marginal zone B cells and weak in germinal center B cells. In B-cell malignancies, CD22 expression ranges from 60% to 80% depending on the histological type and on the assays used. The function of the CD22 molecule is uncertain, although recent studies have suggested roles for the molecule both as a component of the B-cell activation complex and as an adhesion molecule. CD22 deficient mice have a reduced number of mature B cells in the bone marrow and circulation; the B cells have a shorter lifespan and enhanced apoptosis, thus indicating a key role of this antigen in B-cell development/survival. After binding with its natural ligand(s) or antibodies, CD22 is rapidly internalized; this provides a potent costimulatory signal in primary B-cell and proapoptotic signals in neoplastic B cells. Preclinically CD22 has been shown to be an effective target for immunotherapy of B-cell malignancies using either "naked" or toxin-labeled or radiolabeled monoclonal antibodies. Clinical trials in patients with non-Hodgkin's lymphoma (NHL) (both indolent and aggressive disease) are now ongoing with a humanized naked anti-CD22 antibody (epratuzumab, Amgen Inc, thousand Oaks, CA and Immunomedics Inc, Morris Plains, NJ) used as single agent or in combination with other monclonal antibodies (ie, rituximab) and/or chemotherapy. Preliminary data from these studies showed these approaches to be effective and well-tolerated. PMID- 12720148 TI - Radioimmunotherapy for Waldenstrom's macroglobulinemia. AB - Radioimmunotherapy targeting CD20 is a promising novel treatment for lymphoma. Prior trials have established the safe dose of Zevalin ((90)Y-ibritumomab tiuxetan; IDEC Pharmaceuticals) for patients with no more than 25% bone marrow (BM) involvement. Zevalin is expected to be an effective treatment for WM; however, the safe dose has not been defined. A phase I clinical trial has been designed to define the maximum-tolerated dose (MTD) of Zevalin in patients with WM and BM involvement up to 50%. Eligible patients need to have adequate hematologic indices (absolute neutrophil count [ANC] > 1,500/microL, platelets > 100,000/microL). The starting dose of (90)Y-Zevalin is 0.08 mCi/kg. Dose escalation by 0.04 mCi/kg in cohorts of three to six patients will be performed. Patients will be re-treated at 12 weeks if there is no complete response, no progression, and no dose-limiting toxicity. If the degree of BM involvement remains in the 20% to 50% range, re-treatment will involve a similar dose of Zevalin; if it is <20%, patients will receive Zevalin to the maximum allowed cumulative dose of 0.4 mCi/kg (or 0.3 mCi/kg for mild thrombocytopenia). Despite the phase I design, the re-treatment provision is expected to result in significant clinical benefit. PMID- 12720149 TI - Angiogenesis in Waldenstrom's macroglobulinemia. AB - Angiogenesis has been associated with disease progression and poor prognosis in several hematologic malignancies including multiple myeloma. In this article, we summarize the rationale for studying angiogenesis in plasma cell disorders including Waldenstrom's macroglobulinemia (WM). We also discuss the results of a study of angiogenesis in WM conducted at the Mayo Clinic. Our study shows that only 30% of patients with WM had increased (intermediate- or high-grade) angiogenesis. A weak correlation was found between mean microvessel density (MVD) and marrow infiltration. Unlike multiple myeloma, MVD and angiogenesis grade were not predictive factors for survival. Although these findings suggest that angiogenesis may not be a major factor in WM, it does not exclude a paracrine role for angiogenic cytokines or the study of antiangiogenic agents in this disease. PMID- 12720150 TI - Treatment of Waldenstrom's macroglobulinemia with single-agent thalidomide or with the combination of clarithromycin, thalidomide and dexamethasone. AB - To evaluate the activity of thalidomide in Waldenstrom's macroglobulinemia (WM), 20 patients were treated on a dose schedule that escalated from 200 mg/d to 600 mg/d. On an intention-to-treat basis, five (25%) patients achieved a partial response, which was noted within 3 months of treatment. Adverse effects were common and prevented dose escalation of thalidomide in 75% of patients and led to premature discontinuation of treatment in 35%. We subsequently evaluated the oral combination of clarithromycin (500 mg twice per day), low-dose thalidomide (200 mg once daily), and dexamethasone (40 mg once per week). Our preliminary analysis on 12 previously treated patients indicate activity of this regimen in WM: three patients achieved a partial response and two patients demonstrated monoclonal protein reduction of greater than 25%. This combination was associated with a variety of side effects due not only to thalidomide, but also to corticosteroids and to clarithromycin. Our preliminary data indicate that this combination may be a useful salvage regimen for some patients with heavily pretreated macroglobulinemia. PMID- 12720151 TI - Treatment of Waldenstrom's macroglobulinemia with clarithromycin, low-dose thalidomide, and dexamethasone. AB - Twelve patients with Waldenstrom's macroglobulinemia (WM) underwent treatment with the nonmyelosuppressive combination regimen BLT-D: clarithomycin (Biaxin [BXN], Abbott Laboratories, Abbott Park, IL) 500 mg orally twice daily, low-dose thalidomide (THAL) 50 mg orally escalated to 200 mg daily, and dexamethasone (DXM) 40 mg orally once weekly all with modification for toxicity. Omeprazole (correction of omepraxole) 20 mgm orally twice daily for 2 days with the DXM, and enteric-coated aspirin 81 mg orally daily were also administered. Twelve patients have been evaluated. All had previously received at least one purine analogue or alkylating agent. Five had a reduction in either leukocytes and/or platelets prior to treatment, of which three were disease-related. Median age was 62 years. All patients received a minimum of 6 weeks of therapy. Of the 12 patients, 10 had a significant response (83%) consisting of three near complete, three major, four partial, and two minor responses. Four of five had restoration of reduced blood counts. Two with minor responses did not receive sufficient dose escalation due to toxicity. Median time on therapy was 7 months (range, 3 to 28 months). Patients were removed from therapy primarily due to neurotoxicity. Drug resistance occurred in three patients, with one transformation to large cell lymphoma. Toxicity was as follows: gastrointestinal (primarily constipation), 42%; neurological, 100%; endocrine, 42%, and thrombotic, 8%. Most toxicities were World Health Organization (WHO) grade 1 or 2; however, neurological toxicity was more prominent and severe in WM patients than in myeloma. BLT-D is effective in WM. Because of its toxicity, predominantly neurological, BLT-D may best serve as an induction regimen or to "rescue" patients with refractory disease or disease related low counts. PMID- 12720152 TI - Potential new therapeutics for Waldenstrom's macroglobulinemia. AB - Thalidomide the first commercially available immune modulatory drug (IMiD), has activity in the treatment of Waldenstrom's macroglobulinemia (WM), as well as multiple myeloma, myelodysplastic syndrome, myelofibrosis with myeloid metaplasia, chronic lymphocytic leukemia (CLL), and B-cell lymphomas. Although its molecular mechanisms of action have not yet been elucidated, thalidomide and the IMiDs affect a variety of cytokines and inflammatory mediators including tumor necrosis factor-alpha (TNFalpha), interleukin (IL)-1beta, interferon gamma (IFNgamma), IL-6, IL-10, IL-12, and COX-2 and angiogenesis factors such as vascular endothelial growth factor (VEGF) and its receptor. The IMiDs also affect adhesion molecules such as ICAM-1, ICAM-2, and L-CAM, in addition to preferentially stimulating CD8 cells and expanding natural killer (NK) cell populations. Since most IMiDs share these properties, it would be expected that the second-generation IMiDs (REVIMID, ACTIMID) would have activity similar to thalidomide in WM with an improved safety profile. TNFalpha and angiogenesis most likely play a role in promoting the growth and development of WM. The selective cytokine inhibitory drugs (SelCIDs) are potent phosphodiesterase 4 (PDE-4) inhibitors that inhibit TNFalpha production and are highly antiangiogenic. In addition, inhibition of PDE-4 induces apoptosis in human CLL lymphocytes. It is therefore expected that the SelCIDs might have activity in Waldenstrom's tumors. Jun N-terminal kinase (JNK) is a component of signaling cascades that modulate apoptosis, the induction of an inflammatory response via the AP-1 pathway, and modulation of cellular proliferation. In a variety of tumors, including multiple myeloma, JNK is induced as part of a protective mechanism. It is hypothesized that inhibition of JNK activity might allow other chemotherapeutic agents to be more effective in a similar manner to corticosteroids. Work is in progress to evaluate this. Inhibitors of the E3 subunit of ubiquitin ligase may also selectively modulate the expression of receptors, growth factors, and transcription factors essential to the growth, survival, and spread of tumors. We hypothesize that the IMiDs, SelCIDs, JNK inhibitors, and ligase inhibitors will be the basis for a new nonchemotherapeutic approach to the treatment of WM and other related diseases. PMID- 12720153 TI - Role for high-dose therapy with autologous hematopoietic stem cell support in Waldenstrom's macroglobulinemia. AB - Despite effectiveness of standard chemotherapy regimens, complete responses are infrequent in Waldenstrom's macroglobulinemia (WM) and there are no cures. Since WM shares certain biological and clinical features with myeloma, including responsiveness to alkylating agents, evaluation of high-dose therapy (HDT) with transplant, which is effective in myeloma, is an obvious next step in an effort to achieve high response rates and improve survival. Due to the indolent nature of the disease and older patients with comorbidities, such evaluations have been infrequent in the past. We have evaluated the safety and efficacy of high-dose melphalan with peripheral blood stem cell (PBSC) support in eight patients between the ages of 45 and 69 years with WM. Adequate numbers of stem cells were collected in six patients; however, two patients with more extensive prior fludarabine therapy failed to collect adequate cells and required a second attempt at stem cell collection. Seven patients were treated with melphalan 200 mg/m(2), including two patients who received tandem transplants; one patient received melphalan 140 mg/m(2) with total body irradiation (TBI). There was no treatment-related mortality and toxicities were manageable. Recovery of bone marrow after transplant was prompt except in one patient with extensive prior use of fludarabine. All the eight patients achieved at least partial response (PR), including one complete response (CR). Five patients are alive and with out relapse (77+ to 6+ months post-transplant). Other investigators have reported similar experience suggesting safety and efficacy of HDT in WM. However, therapy with purine analogues leads to stem cell damage with decreased ability to collect adequate numbers of stem cells. This suggests that the PBSCs should preferably be procured prior to extensive use of purine analogues. Future strategies in WM will include a plan to evaluate the role of HDT along with biological agents, role of purging using rituximab, post-HDT maintenance strategies (including immunotherapy), and evaluation of nonmyeloablative regimens containing fludarabine to achieve higher response rates and improve survival. PMID- 12720154 TI - Autologous and allogeneic stem cell transplantation in Waldenstrom's macroglobulinemia: review of the literature and future directions. AB - Waldenstrom's macroglobulinemia (WM) is a chronic lymphoproliferative disorder characterized by lymphoplasmacytic infiltrate and a monoclonal IgM serum peak. Treatment includes cytotoxic chemotherapy with alkylators, or purine nucleoside analogues and monoclonal anti-CD20 antibody. The role of stem cell transplantation (SCT) in WM has not been established. We identified 24 published cases of WM treated with high-dose chemotherapy (HDC) followed by autologous SCT (ASCT). The median age was 50 years; half of the patients had refractory disease and received a variety of preparative regimens. Nine complete and 14 partial responses were observed, with one early death. Fifteen patients were alive and well at follow-ups ranging from 1 to 132 months. Six additional patients, with a median age of 45 years (range, 30 to 62), who received allogeneic SCT have been reported. All were heavily pretreated with refractory or relapsed disease. Median time from diagnosis to transplant was 3.1 years (range, 1.3 to 7). Two patients died of complications of the procedure while one died of disease progression. Three patients were alive and well between 5 and 112 months post-transplant. The small number of reported patients precludes any significant conclusion except that SCT is feasible in WM and long-term disease control can be achieved in selected patients with autologous or allogeneic SCT, even in those with refractory disease. PMID- 12720155 TI - Transplantation in Waldenstrom's macroglobulinemia--the French experience. AB - Published data on transplantation in Waldenstrom's macroglobulinemia (WM) are still limited. We present a retrospective multicentric study of 27 WM patients who underwent 19 autologous (median age, 54 years) and 10 allogeneic (median age, 46 years) transplantations. Median time between diagnosis and transplantation was 36 months; 66% of patients had received three or more treatment lines and 72 % had chemosensitive disease. High-dose therapy (HDT) and autologous transplantation induced a 95% response rate (RR), including 10 major responses. With a median follow-up of 18 months, 12 patients are alive at 10 to 81 months and eight are free of disease progression at 10 to 34 months. The toxic mortality rate (TRM) was 6%. Allogeneic transplantation was preceded by HDT in nine patients and by a nonmyeloablative regimen in one patient. The RR was 80%, including seven major responses. With a median follow-up of 20.5 months, six patients are alive and free of progression at 3 to 76 months. Four patients died, all from toxicity, resulting in a TRM of 40%. HDT followed by autologous transplantation is feasible in WM, even in heavily pretreated patients, with some prolonged responses but a high relapse rate. Conversely, allogeneic transplantation is more toxic, but likely induces a graft-versus-WM effect and may, for some patients, result in long-term disease control. PMID- 12720156 TI - Modulation of the activity of Bcl-2 in Waldenstrom's macroglobulinemia using antisense oligonucleotides. AB - Bcl-2 is an attractive target for anticancer therapy in a number of malignancies, as its expression is associated with inhibition of the apoptotic program and resistance to traditional therapeutic agents. Bcl-2 antisense therapy with G3139 (oblimersen sodium; Genasense, Genta Inc, Berkeley Heights, NJ) is in clinical trials for a number of malignancies, including an ongoing trial in myeloma. In vitro G3139 has been shown to downregulate Bcl-2 in myeloma cells, sensitizing them to chemotherapeutic agents. We have undertaken a project to evaluate antisense inhibition strategies in Waldenstrom's macroglobulinemia (WM), and whether the Bcl-2 pathway may provide a therapeutic target in this disease. We have shown that Bcl-2 is expressed in WM cells in vitro and that downregulation of Bcl-2 may be achieved by treatment with G3139. Treatment of WM cells with G3139 is associated with increased cell death and shows potential synergy with chemotherapeutic agents active in WM. Bcl-2 downregulation via G3139 antisense treatment may have potential anticancer efficacy in WM and further studies to address its effects on clinical specimens are warranted, in anticipation of using this agent in WM clinical trials. PMID- 12720157 TI - Oblimersen sodium (G3139 Bcl-2 antisense oligonucleotide) therapy in Waldenstrom's macroglobulinemia: a targeted approach to enhance apoptosis. AB - The components of the apoptotic pathway are targets for anticancer therapy. Bcl-2 protein inhibits apoptosis and confers resistance to treatment with traditional cytotoxic chemotherapy, radiotherapy, and monoclonal antibodies. Oblimersen sodium (G3139, Genasense, Genta Inc, Berkeley Heights, NJ) is an antisense oligonucleotide compound designed to specifically bind to the first six codons of the human bcl-2 mRNA sequence, resulting in degradation of bcl-2 mRNA and subsequent decrease in Bcl-2 protein translation. Oblimersen is the first oligonucleotide to demonstrate proof of principle of an antisense effect in human tumors by the documented downregulation of the target Bcl-2 protein. A growing body of preclinical and clinical evidence suggests that oblimersen synergizes with many cytotoxic and biologic/immunotherapeutic agents against a variety of hematologic malignancies and solid tumors. Randomized clinical trials are currently underway to evaluate the efficacy and tolerability of oblimersen in combination with cytotoxic chemotherapy in chronic lymphocytic leukemia (CLL), multiple myeloma (MM), malignant melanoma, and non-small cell lung cancer. In addition, nonrandomized trials are underway to evaluate oblimersen in non Hodgkin's lymphoma (NHL), acute myeloid leukemia (AML), and hormone-refractory prostate cancer. Preclinical data support the clinical evaluation of oblimersen in additional tumor types, including chronic myelogenous leukemia, and breast, small cell lung, gastric, colon, bladder (CML), and Merkel cell cancers. Enhancement of the efficacy of anticancer treatments with oblimersen Bcl-2 antisense therapy represents a promising new apoptosis-modulating strategy, and ongoing clinical trials will test this therapeutic approach. PMID- 12720158 TI - Dendritic cell-mediated immunization in macroglobulinemia. AB - Dendritic cells (DCs) are professional antigen-presenting cells (APCs) specialized to initiate immunity and are critical targets for all vaccines. New methods to generate large quantities of these cells in the laboratory and load them with tumor antigens allows novel approaches targeting these APCs in the clinic. Injection of antigen loaded DCs can boost tumor-specific immunity in patients. Studies in myeloma suggest that the use of autologous tumor as a source of antigen may be the preferred approach. Boosting antitumor immunity may be a useful strategy to prevent tumor progression in patients with asymptomatic macroglobulinemia. PMID- 12720159 TI - Novel biologically based therapies for Waldenstrom's macroglobulinemia. AB - Waldenstrom's macroglobulinemia (WM) remains an incurable B-cell malignancy, necessitating urgent development of novel treatment strategies. Building on our experience on bed-to-bedside translational studies for multiple myeloma (mm), we identified a constellation of novel classes of anti-WM agents, including the proteasome inhibitor PS-341; the ansamycin family of inhibitors (eg, geldanamycin and its analogues) of the heat-shock protein 90 (hsp90) molecular chaperone; histone deacetylase inhibitors, such as suberoylanilide hydroxamic acid (SAHA); and the thiazolidinedione group of peroxisome proliferator-activated receptor gamma (PPAR-gamma) agonists (eg, ciglitazone or rosiglitazone). Our preclinical data show that these classes of agents induce growth arrest and apoptosis of WM cells, at concentrations relevant to those achieved in previous clinical uses of these drugs, and suggest that novel therapeutic strategies for WM can be designed to include combinations of these agents, to simultaneously target multiple levels of diverse pathways important for tumor cell growth and survival, and thus maximize the pro-apoptotic activities of these agents and/or neutralize protective responses of WM against their effects. These molecular studies provide a framework for rational design of the next generation of combination therapies for WM. PMID- 12720160 TI - The Wayne State University Waldenstrom's Macroglobulinemia preclinical model for Waldenstrom's macroglobulinemia. AB - The Wayne State University Waldenstrom's Macroglobulinemia xenograft model in mice with severe combined immune deficiency (WSU-WM-SCID) is the only preclinical animal model available for this disease. It is based on a permanent, EBV- IgMlambda cell line (WSU-WM) established from a patient with a 10-year history of Waldenstrom's macroglobulinemia (WM). These cells are CD5( )CD10(+)CD19(+)CD20(+)CD22(+) and have t(8;14) (q24;32), t(12;17) (q24;q21), 2p-. WSU-WM cells also express DNA topoisomerase II (alpha and beta), and are bcl(2)(+)bcl(XL)(+)bax(-). Although the tumor has aggressive biological behavior with c-myc-IgH rearrangement, it has retained the salient features of WM. The breakpoint on 8q24 is downstream of c-myc exon 3, which is not usual for Burkitt type breakpoints. WSU-WM cells also express both secretory (s(u)) and membrane (m(u)) IgM mRNA and secrete IgM in culture supernatant. Histiologically, WSU-WM SCID xenograft tumors have lymphoplasmacytoid morphology. These features indicate biological, but not histological evolution. The WSU-WM-SCID is a model of a more aggressive and resistant WM usually seen toward the late stages of disease. It is, therefore, a particularly useful tool in developing new therapeutic strategies for the more aggressive WM, including targeted therapy, which exploits unique molecular characteristics of tumor cells. PMID- 12720161 TI - Human monoclonal macroglobulins with antibody activity. AB - Assays for specific antigen-binding activity were performed on sera from 172 patients with monoclonal macroglobulinemia defined by immunofixation electrophoresis. The sera were collected between 1970 and 2002. Mean IgM level was 1,409 mg/dL with a range from 70 to 6,800. Cryoglobulins were identified in 15.3% (26/170 sera: 12 trace, five single component, and nine mixed IgM-IgG). Rheumatoid factor (RF) was detected in 19 of 151 (12.6%) samples with titers ranging from 1:80 to 1:327,680. Among the nine mixed IgM-IgG cryos, eight were RF positive and six of six displayed positivity for hepatitis C virus. Cold agglutinins (CA) were present in 8.5% (10/117) of sera with anti-I titers between 1:512 and 1:65,536. IgM binding to a series of glycosaminoglycan oligosaccharides, glycolipids, and glycoprotein antigens was found in 75 samples (43%). IgM binding to antigens having known associations to polyneuropathies occurred in 20 patients (12%). Antinuclear antibody (ANA) was documented in 10.7% (18/169) of sera. Anti-DNA activity was absent in all samples tested. Sera from 71% of patients with monoclonal macroglobulinemia in this series exhibited binding to autoantigens. Some of these immune complexes resulted in clinically significant manifestations. Our results suggest that many monoclonal immunoglobulins may be functional antibodies rather than "paraproteins." Characterization of antigen-binding activities may provide insight into the pathogenesis of monoclonal gammopathies. PMID- 12720162 TI - Amyloidosis with IgM monoclonal gammopathies. AB - We sought to review outcomes of patients who have immunoglobulin light-chain amyloidosis of the IgM type. Fifty patients with a serum IgM monoclonal protein and biopsy-proven amyloid were evaluated. The percentages of patients presenting with cardiac, renal, hepatic, and pulmonary amyloid were 44%, 32%, 14%, and 10%, respectively. Forty-two percent had an M-protein spike in the serum greater than 1.5 g/dL, and 14% had an IgM peak greater than 3 g/dL. Amyloidosis was diagnosed easily by biopsies of the subcutaneous fat, rectum, and bone marrow, which demonstrated deposits in 84%, 72%, and 50%, respectively. The median survival of all patients was 24.6 months. Fifty-three percent of all deaths were due to cardiac amyloidosis. Twelve percent died of respiratory failure, and 7% each died of macroglobulinemia, hepatic failure, and renal failure. Of the 22 patients with amyloid cardiomyopathy, the median survival was 11.1 months and fewer than 10% survived 5 years. Of the 28 patients without amyloid cardiomyopathy at diagnosis, the median survival was 27 months, with approximately 30% surviving 5 years. We conclude that the presence of amyloid cardiomyopathy and an increased creatinine concentration had the greatest impact on survival. PMID- 12720163 TI - Appropriateness of applying the response criteria for multiple myeloma to Waldenstrom's macroglobulinemia? AB - Since the presence of an M-component is an essential disease feature in both multiple myeloma (MM) and Waldenstrom's macroglobulinemia (WM), the decrease in the M-protein size applied for response in MM is also a crucial criteria for assessing response in WM. However, WM frequently displays lymphoma-like features that should be included in the response criteria. PMID- 12720164 TI - Increased CT scan utilization does not improve the diagnostic accuracy of appendicitis in children. AB - BACKGROUND/PURPOSE: Appendicitis continues to present a diagnostic dilemma in children of all ages leading to increased utilization of radiographic studies. Focused computed tomography (CT) scanning has become the diagnostic test of choice in many hospitals. The purpose of this study was to critically evaluate the use of radiographic studies for the evaluation of acute appendicitis in children and to determine if diagnostic accuracy has improved. METHODS: Children undergoing appendectomy for acute appendicitis were reviewed from 1997 to 2001. Diagnostic workup (CT scan, ultrasound [US], or no radiographic study) was recorded as were the final pathology results. RESULTS: Six hundred sixteen appendectomies were performed. Mean age was 10.4 +/- 4.1 years, and 60% were boys. Overall, 184 children (30%) underwent CT scanning, 104 (17%) had US performed, and 310 (50%) had no radiographic study (18 patients had both CT and US performed). A pathologically normal appendix was removed in 7% (14 of 202) of CT patients, 11% (14 of 122) of US patients, and 8% (26 of 310) of patients without a study. The frequency of CT scanning increased from 1.3% of all children in 1997 to 58% in 2001, whereas utilization of US decreased from 40% to 7%. Over the same period, the overall negative appendectomy rate did not change significantly from 8% to 7%. CONCLUSIONS: With increased utilization of focused CT scanning, the negative appendectomy rate has remained unchanged. History and physical examination by an experienced surgeon is as accurate as CT in correctly diagnosing acute appendicitis in children. PMID- 12720165 TI - Pediatric surgical emergencies in the setting of a natural disaster: Experiences from the 2001 earthquake in Gujarat, India. AB - BACKGROUND/PURPOSE: On January 26, 2001, a 7.9 Richter earthquake struck the Indian state of Gujarat. Over the next 6 days, the International Red Cross set up a mobile hospital in the city of Bhuj, near the epicenter. The authors describe all surgeries on children treated there during the first 4 weeks of operation. The evolution of presenting injuries is noted, the types of surgery required are classified and an effective disaster relief team composition and strategy are proposed. METHODS: Total casualties were estimated at 30,000, with 250,000 people injured. Of 1,142 inpatients treated at Nor-Finn hospital during the first 4 weeks, approximately 300 (25%) were /=11 years). GID location by frequency was ileum (31.5%), ileocaecal valve (30.2%), duodenum (9.6%), stomach (8.2%), jejunum (8.2%), colon (6.8%), and rectum (5.5%). In neonates and infants, vomiting and distension were the most common presentations. Volvulus, caused by a duplication, occurred in 23.8% of neonates and caused the death of one neonate. Intussusception was identified in 10.9% of patients. In older children and adolescents, pain and vomiting were the most common associations. Six of these patients were being treated for Crohn's disease, with the diagnosis of duplication made at laparotomy. Eighteen patients had a prenatal diagnosis by ultrasound scan, with 77.2% of these asymptomatic after birth. Most prenatal diagnoses occurred after 1991 (77.8%). When comparing an earlier period (1980 to 1991; 29 patients) with the current (1992 to 2002; 44 patients), a greater proportion of the latter patients were asymptomatic (36.4 v 13.8%) and had a lower incidence of complications (volvulus/intussusception). CONCLUSIONS: GID can lead to life threatening complications. Prenatal diagnosis should lead to expeditious postnatal investigation and treatment before the onset of symptoms or complications. GID in older children can mimic Crohn's disease. Laparoscopy/laparotomy should be considered in patients with atypical Crohn's disease or when the diagnosis of an intraabdominal mass is unclear. PMID- 12720185 TI - Image-guided percutaneous approach is superior to the thoracoscopic approach in the diagnosis of pulmonary nodules in children. AB - BACKGROUND/PURPOSE: Image-guided, percutaneous techniques are increasingly used in diagnosis of pulmonary disease in children. The aim of this study was to determine the diagnostic accuracy and clinical outcomes of thoracoscopic versus percutaneous lung biopsy in children. METHODS: Sixty-three consecutive patients from January 1996 to December 2000 who had a thoracoscopic lung biopsy, a percutaneous ultrasound scan, or computed tomography (CT)-guided lung biopsy for well-defined and ill-defined lesions were analyzed. RESULTS: Twenty-eight patients had a thoracoscopic lung biopsy (TLB), and 35 patients had a percutaneous image-guided lung biopsy (PLB). Age ranged from 6 months to 17 years (median, 8 years). There was no significant difference between groups with regard to age, depth of lung nodule biopsied, or prebiopsy diagnoses. Seventeen patients (60%) of TLB and 23 (65%) of PLB had well-defined pulmonary nodules suspicious for malignancy at the time of biopsy. Adequate tissue for pathologic diagnosis was obtained in 28 (100%) of TLB versus 26 (80%) of PLB patients. However, 8 (28%) thoracoscopic cases needed to be converted to an open procedure. In 3 (8.5%) PLB cases the percutaneous biopsy was insufficient, and a thoracoscopic or open biopsy was required. The median hospital stay was 3 days for TLB and 4 to 6 hours for PLB (P =.023). There were no complications in the PLB group. Five (18%) of TLB patients suffered a persistent air leak treated with continued chest tube drainage, and one patient died of other causes with a persistent air leak. CONCLUSIONS: Percutaneous lung biopsy has a significantly shorter hospital stay and a lower complication rate than thoracoscopic lung biopsy. The authors propose that the percutaneous technique should be considered as the initial approach for children with pulmonary nodules. PMID- 12720186 TI - Alterations in smooth muscle contractile and cytoskeleton proteins and interstitial cells of Cajal in megacystis microcolon intestinal hypoperistalsis syndrome. AB - BACKGROUND/PURPOSE: Megacystis microcolon intestinal hypoperistalsis syndrome (MMIHS) is characterized by decreased or absent peristalsis. Gastrointestinal motility depends on the enteric nervous system, smooth muscle cells (SMCs), and the interstitial cells of Cajal (ICCs). Contractile and cytoskeleton proteinase are important structural and functional components of SMCs. The aim of study was to examine the expression of contractile and cytoskeleton proteins in SMCs and distribution of ICCs in MMIHS bowel. METHODS: Full-thickness bowel specimens were obtained from 4 infants with MMIHS and 4 controls. Specimens were processed as whole-mount preparations and frozen and paraffin sections. Combined staining of NADPH-d histochemistry/c-kit immunohistochemistry, single and double immunohistochemistry using alpha-smooth muscle actin (alpha-SMA), calponin (CALP), caldesmon (CALD), desmin (DES), protein gene product 9.5 (PGP 9.5) and c kit antibodies were performed and examined using light and confocal scanning microscopy. RESULTS: alpha-SMA, CALP, CALD, and DES immunoreactivity were reduced markedly in MMIHS bowel compared with controls. Combined NADPH/c-kit staining showed dense network of ICCs around myenteric plexus in MMIHS bowel. In contrast, the intramuscular ICCs either were absent or reduced in MMIHS bowel. CONCLUSIONS: Marked reduction of contractile and cytoskeleton proteins in SMCs combined with reduced expression of intramuscular ICCs in the gut may be responsible for the motility dysfunction in MMIHS. PMID- 12720187 TI - Is contralateral exploration necessary in girls with unilateral inguinal hernia? AB - BACKGROUND/PURPOSE: Routine contralateral groin exploration in girls with unilateral inguinal hernia (UIH) continues to be controversial. The aim of this study was to determine the incidence of contralateral hernia development in girls after UIH repair. METHODS: Between 1972 and 2000, 391 girls underwent repair of UIH. Ninety-one (23%) of the 391 girls underwent routine contralateral exploration during UIH repair and were excluded from the study. The median age at operation in the remaining 300 girls was 3.3 years (range, 1 month to 14 years). Two hundred ten (70%) girls had right-sided and 90 (30%) had left-sided UIH. Familial history of hernia was identified in 8 (3%) patients. The follow-up ranged from 1.5 to 14 years. Mann-Whitney U test was used for intergroup comparison. RESULTS: A contralateral hernia developed in 24 (8%) of the 300 patients who had unilateral repair of inguinal hernia. Median time from operation to occurrence of the contralateral hernia was 3 years (range, 1 to 4 years). Age at operation, side of hernia, and familial history did not influence the development of contralateral hernia. CONCLUSIONS: These data suggest that the low incidence of contralateral hernia development in girls undergoing UIH repair does not justify routine contralateral groin exploration. PMID- 12720188 TI - Anal canal duplication in infants. AB - BACKGROUND/PURPOSE: Anal canal duplication (ACD) is the most distal and the least frequent digestive duplication. A review of the English-language literature found 15 cases reported in the pediatric age group. METHODS: A retrospective chart review was performed for our experience from 1999 to 2001 with 6 patients who presented with a midline postanal opening. RESULTS: All of 6 patients were girls and included one set of twins. Patients were from 3 to 9 months of age at the time of diagnosis (mean, 4.5 months). The anal canal duplication was delineated clearly by contrast study of the tract. All were tubular structures, 10 to 12 mm in length. None of the patients had any other associated anomalies. Five of 6 patients underwent operation between the ages of 3 and 8 months (mean, 5.4 months). Excision of the ACD was accomplished through the posterior sagittal approach. The orifice of the ACD, measuring 1 to 2 mm in diameter located just behind anus, directed to the lumen of the anal canal by keeping in the midline and ended blindly 5 mm above the dentate line without luminal communication. Histology findings showed a squamous epithelium with smooth muscle bundles in 2 cases and pseudostratified columnar epithelium with focally squamous epithelial lining and adjacent smooth muscle bundles in 3 cases. The postoperative courses were uneventful with satisfactory anal function. One patient has not yet undergone operation and has been well on outpatient follow-up. CONCLUSIONS: ACD is a congenital developmental lesion located in the midline posterior to the anus presenting as a tubular structure without communication with the anal canal, usually discovered in early infancy, and characteristically predominant in girls. The authors recommend that all ACDs, regardness of size and length, should be removed surgically to restore the normal anatomy and to avoid delayed presentation of infection such as perianal abscess or fistula formation. PMID- 12720189 TI - Are localized intestinal perforations distinct from necrotizing enterocolitis? AB - BACKGROUND/PURPOSE: Localized intestinal perforation (LP) is thought to be a distinct entity when compared with perforation associated with necrotizing enterocolitis (NEC). Studies have indicated that LP is more amenable to percutaneous drainage and associated with a better outcome. We sought to determine whether LP and NEC could be distinguished based on clinical parameters alone. METHODS: A retrospective review of 40 neonates with gastrointestinal perforations between January 1990 and May 1998 was performed. All had undergone laparotomy and had histologic specimens available for evaluation. RESULTS: Twenty one neonates had necrotizing enterocolitis (NEC), and 19 had localized perforation (LP) based on histologic criteria. More neonates with LP were exposed to prenatal indomethacin (37% v 5%; P <.05), received intravenous dexamethasone (42% v 10%; P <.05), had umbilical artery catheters (63% v 14%; P <.05), and had a higher white blood cell (WBC) count (27.1 +/- 23.1 v 14.3 +/- 11.5; P <.05). More neonates with NEC had pneumatosis intestinalis (47% v 11%; P <.05). No significant differences existed in enteral feeding (16% LP v 38% NEC) or overall mortality rate (37% LP v 38% NEC). No statistical differences in the timing of perforation or clinical presentation were found. CONCLUSIONS: NEC and LP are difficult to distinguish based on clinical parameters alone. The authors did find associations between LP and prenatal indomethacin, intravenous dexamethasone, umbilical artery catheters, and a higher WBC count. Mortality rate and clinical outcome were nearly identical in both groups. Pneumatosis intestinalis, thought to be pathognomonic for NEC, was seen on abdominal radiograph in 2 babies with histologically proven LP. PMID- 12720190 TI - A simple technique of laparoscopic full-thickness anterior abdominal wall repair of retrosternal (Morgagni) hernias. AB - BACKGROUND/PURPOSE: Previous reports of laparoscopic repair of Morgagni hernias in children have involved relatively complex laparoscopic techniques. This report describes a simpler method of repair that we have applied to 4 children. METHODS: Four children with retrosternal (Morgagni) hernias underwent primary laparoscopic repair by placement of interrupted synthetic nonabsorbable sutures through the full-thickness of the anterior abdominal wall, incorporating the posterior rim of the defect and returning back out through the anterior abdominal wall, with the sutures tied in the subcutaneous tissue. RESULTS: The children, ranging in age from 11 to 36 months, underwent laparoscopic repair of their Morgagni hernias and had an uneventful postoperative recovery, apart from a port site hernia in one. CONCLUSIONS: This technique for primary laparoscopic repair of Morgagni hernia is easy to perform, well tolerated by the patient, and gives excellent cosmetic results. Laparoscopic closure of the defect by suturing the posterior rim of the hernia to the full thickness of the anterior abdominal wall would appear to provide a safe and effective means of repairing this type of hernia. PMID- 12720191 TI - Gastroschisis: A sixteen-year review. AB - PURPOSE: The aim of this study was to examine the incidence of gastroschisis in Saskatchewan over a 16-year period and identify risk factors associated with mortality. METHODS: Information was obtained by retrospective review of all infant charts at the only 2 provincial neonatal intensive care centers from January 1985 through December 2000. Factors recorded were gestational age, birth weight, gender, Apgar score, time to operation, method of closure, time to attain full feedings, presence of sepsis, and length of hospital stay. Live birth data for the province was obtained, and the incidence per 10,000 live births for 3 time intervals was calculated. Analysis was performed with mortality as the dependent variable. RESULTS: Seventy-one infants were identified. Overall survival rate was 93% (66 of 71). During the 3 time intervals examined, the incidence of gastroschisis per 10,000 live births increased from 1.85 in 1985 to 1990 to 3.66 in 1991 to 1995 to 4.06 in 1996 to 2000. The analysis found that intestinal atresia P =.009, OR = 18.3 (95%CI: 2.457-136) and intestinal necrosis P =.050, OR = 10.33 (95%CI: 1.32-80.68) were significantly associated with mortality. CONCLUSIONS: The incidence of gastroschisis is increasing. Intestinal atresia and the development of intestinal necrosis were associated with poor outcome. Patients who had these complications all had short bowel, and full feedings could not be established. They required continuation of total parenteral nutrition and experienced episodes of sepsis. PMID- 12720192 TI - Correlation between radiographic transition zone and level of aganglionosis in Hirschsprung's disease: Implications for surgical approach. AB - BACKGROUND/PURPOSE: The anticipated level of aganglionosis can influence the surgical approach to Hirschsprung's disease. The aim of this study was to determine the accuracy of the contrast enema in predicting this level. METHODS: Over a 6-year period (1995 through 2000), 88 patients with Hirschsprung's disease underwent surgical correction. Preoperative contrast enema findings were available for 75 of these patients and were compared with operative and pathology reports. Data were analyzed by chi(2). RESULTS: The contrast enema showed a transition zone suggestive of Hirschsprung's disease in 67 of 75 patients (89%). In 59 of 67 (88%), the pathologic and radiographic transition zones were concordant. Seven of the 8 patients with discordant studies had total colonic (n = 5) or long-segment (n = 2) disease. Contrast enema correctly predicted the level of aganglionosis in 55 of 62 (89%) patients with rectosigmoid disease but only 4 of 13 (31%) of those with long-segment or total colonic disease (P <.01). Of the patients with a radiographic transition zone in the rectosigmoid, 54 of 60 (90%) had a matching level of aganglionosis. CONCLUSIONS: In rectosigmoid Hirschsprung's disease, the location of the radiographic transition zone correlates accurately with the level of aganglionosis in 90% of cases. However, the small incidence of discordance between anticipated level of aganglionosis and operative findings should be recognized, particularly when planning a one-stage transanal pull-through. PMID- 12720193 TI - Quality of life of patients who have undergone the Nuss procedure for pectus excavatum: Preliminary findings. AB - BACKGROUND/PURPOSE: The current project is a preliminary qualitative exploration of changes in quality of life of patients who have undergone the Nuss Procedure. The current study explores quality of life after surgical repair from the perspectives of both the patients and the parent(s) of the younger participants. METHODS: This research constitutes the first segment in a mixed-method longitudinal design. The author conducted semistructured interviews based on the Keith and Schalock's quality of life model. Five youth who have undergone the Nuss Procedure and the parent(s) of the 4 younger participants were included in the study for a total of 10 participants. Textual analysis has been carried out using Atlas.ti, a qualitative data analysis program that facilitates such activities as selecting, coding, and comparing textual segments. RESULTS: Results based on these preliminary data indicate that the patients interviewed had significant improvement in overall quality of life, which they attributed to the surgery. Specific examples of patients' expressions of improved self-confidence and renewed interest in physical activity are given. CONCLUSIONS: The Nuss Procedure is a minimally invasive surgical option for correcting pectus excavatum deformities that may make important contributions to patients' self-perceptions and quality of life. The participants in the current study expressed satisfaction with both the physical results of the surgery and with how it improved their overall quality of life. The authors recommend further prospective longitudinal research that assesses pre- and postsurgery psychosocial status and the maintenance of perceived quality of life changes. PMID- 12720194 TI - Snowmobile injuries and fatalities in children. AB - BACKGROUND/PURPOSE: Snowmobiling is a popular form of wintertime recreation but can be associated with significant morbidity and mortality. To better understand snowmobile trauma in children, medical records were reviewed, evaluating the relationships between demographic data, mechanisms, and resultant injuries. In addition, because prior studies of childhood snowmobile fatalities have reviewed only national databases, state and national data were combined to evaluate possible underreporting. METHODS: Medical records were reviewed of children .05). In our institution, the cost of a TCT is $200, and the patient charge is $906 ($94 per CXR). Based on our study data 200 TCTs would need to be done for each clinically significant change, increasing patient ($180,000) and hospital ($39,600) costs. CONCLUSIONS: Helical TCT is a highly sensitive imaging modality for the thoracic cavity; however, routine CXR still provides clinically valuable information for the initial trauma evaluation at minimal cost. TCT should be reserved for selected cases and not as a primary imaging tool. PMID- 12720197 TI - Surgical implications of urachal remnants: Presentation and management. AB - BACKGROUND/PURPOSE: The abdominal manifestations of urachal remnants often prompt referral to the pediatric general surgeon. The purpose of this study was to evaluate the authors' management of this anomaly. METHODS: The authors performed a retrospective review of patients presenting to their institution with urachal remnants between 1984 and 2001. Clinical and radiographic details of presentation, management, and outcomes are described. RESULTS: Twenty-six patients presented at a median age of 4 years (range, 2 days to 12 years), 16 were boys, and 18 required inpatient care. Eleven (42%) presented with infection, 7 (27%) with clear drainage, 3 (12%) with umbilical polyps/granulation, 3 (12%) with pain, one (4%) with recurrent urinary tract infections, and one (4%) with an asymptomatic punctum. One had an associated anomaly (hypospadias). Urinalysis and urine cultures did not correlate with infection. Ultrasound scan was diagnostic in greater than 90% of cases. Overall, 20 patients underwent primary cyst excision, and 6 underwent incision and drainage (I&D) with delayed excision. Five patients underwent primary excision while infected, and 2 had postoperative complications (wound infection and urine leak). All 6 patients who underwent 2 stage procedure initially presented with infection, and none had complications. CONCLUSIONS: Persistent urachal remnants can present at any age with a variety of clinical manifestations. Ultrasound scan is a reliable diagnostic tool. Additional diagnostic studies generally are not warranted. Simple excision of noninfected lesions is appropriate. In cases of acute infection, initial I&D with delayed cyst excision may be preferable to avoid unnecessary complications. PMID- 12720198 TI - Life on the farm-children at risk. AB - BACKGROUND: More than 1 million children live, play, and work on farms, surrounded by animals and machinery. This symbiotic relationship between work and home exposes children to unique risks. METHODS: Children presenting with a farm related injury (November 1994 to August 2001, 82 months) were included. Trauma registry parameters included injury severity score (ISS); Glascow Coma Scale (GCS); time to presentation; season and day of injury; emergency room, intensive care unit, and total length of stay type; and mechanism of injury; and operations. RESULTS: A total of 1,832 pediatric trauma patients were evaluated. Ninety-four children were identified with farm-related injuries. Mean age was 10.75 years. Mean ISS was 7.38. Three children died. Four children wore protective equipment. Forty-four percent of injuries occurred during summer, 31% during spring, and 55% on weekends. Average time to initial presentation was 39 minutes. A total of 177 minutes elapsed before transfer to regional trauma center. Seventy-two children required admission. LOS was 0 to 28 days, mean, 2.76 days. Twenty-six children (28%) required operations. Injuries included dislocations/fractures (52%), lacerations/avulsions (38%), concussions (31%), contusions (30%), and burns (14%). Mechanism included animals (41%), falls (34%), motor vehicles (28%), all-terrain vehicles (20%), and firearms (4%). CONCLUSIONS: Farm injuries occur most commonly during weekends, summer, and spring months, resulting in significant morbidity. Most injuries required hospitalization. Unless unstable, initial transfer to a regional pediatric trauma center should result in the most cost-effective, prompt, and highest quality of care. PMID- 12720199 TI - Expression of heme oxygenase-1 and endothelial nitric oxide synthase in the lung of newborns with congenital diaphragmatic hernia and persistent pulmonary hypertension. AB - BACKGROUND/PURPOSE: Heme oxygenase (HO-1), an inducible isoform of HO is a regulator of vascular tone and cell proliferation through the production of endogenous carbon monoxide (CO). Endothelium-derived nitric oxide (NO) occurs in the endothelial layers of blood vessels and mediates vasorelaxation. Both CO and NO have similar properties and are potent vasodilators. The aim of this study was to examine the expression of HO-1 and endothelial nitric oxide synthase (eNOS) in the Congenital diaphragmatic hernia (CDI) lung. METHODS: RNA was extracted from archival formalin-fixed paraffin-embedded lung tissue from 11 patients with CDH complicated by persistent pulmonary hypertension (PPH). Five age-matched newborns served as controls. Reverse transcription polymerase chain reaction (RT-PCR) was performed using specific primers for human HO-1 and eNOS. Immunohistochemistry using HO-1 and eNOS antibodies was performed and examined using laser scanning microscope. RESULTS: HO-1 and eNOS mRNA expression was significantly decreased in CDH lung compared with controls (P <.05). HO-1 and eNOS immunoreactivity was reduced markedly reduced in the endothelium and arterial wall in the CDH samples compared with normal lung. CONCLUSIONS: Decreased expression of HO-1 and eNOS in the CDH lung suggests deficiency of endogenous NO and CO, which may contribute to altered vascular tone causing PPH. PMID- 12720200 TI - Strictureplasty: An alternative approach in long segment bowel stenosis Crohn's disease. AB - BACKGROUND/PURPOSE: Intestinal resection is the most frequent surgical procedure for bowel stenoses in Crohn's disease (CD). Recurrence of strictures, particularly with ileocolonic disease, often requires resection of lengthy segments of bowel, potentially resulting in short bowel syndrome. Different techniques of strictureplasty, such as those described by Mikulicz, Finney and Michelassi, are used in adults. However, these procedures are uncommon in pediatric surgery. The authors report their experience with different techniques of strictureplasty and with their modified Michelassi technique for the surgical treatment of long intestinal strictures caused by CD. METHODS: Five adolescents (2 boys; 3 girls; mean age, 16 age; range, 14 to 20 years) with severe ileocolonic stenoses and intestinal obstruction, not responsive to medical and nutritional therapy, were treated with different strictureplasty techniques. In 3 of them the modified side-to-side Michelassi technique was used. RESULTS: No postoperative complications occurred. After a mean follow-up of 20.5 months (range, 6 to 28 months), patients are free of symptoms with good nutritional status and off steroid therapy. CONCLUSIONS: Strictureplasty is a good and effective surgical option for sparing bowel length in CD patients with extensive intestinal strictures. PMID- 12720201 TI - Combined approach to functional constipation in children. AB - BACKGROUND/PURPOSE: The authors' 15-year experience with children shows a high percentage of recurrence of functional constipation (FC) with conventional treatment. These data, confirmed in the international literature, led them to develop a new therapeutic approach. The aim of this study was to achieve intestinal control and avoid recurrence of FC in children through use of medical psychological treatment. METHODS: The authors studied 25 children (18 boys; mean age, 4.7 years; range, 2.10 to 7), 20% of whom had anal fissure, 30% retentive soiling, 52% pain on defecation, and 88% fecal retention owing to FC. Children and parents were questioned about eating and sleeping habits, school, toilet training, and daily routine. Treatment included increasing water and fiber intake, laxatives, and family therapy including making rules and working on autonomy and paternal role. RESULTS: Mean onset of FC was 3.5 years, after "stressful events" in 88%. The questionnaire shows that 68% lacked parental autonomy and authority; 84% of children decided on their own about eating habits and sleeping; 68% had a "peripheral" father with a mother-child symbiotic relationship. After one month of therapy, 92% of children showed a modification of at least 2 behavioral patterns; after 3 months, 88% had regular bowel movements. During follow-up (range, 6 to 28 months), 48% had 2 or 3 recurrent episodes. After one year, 68% had reinforced the new behavioral patterns with resolution of the pathologic aspects. CONCLUSIONS: A multidisciplinary approach in the treatment of childhood functional constipation showed consistent therapeutic results by making rules and by equalizing family roles. PMID- 12720202 TI - 'High' anorectal malformation in boys. PMID- 12720204 TI - Pediatric oncology nursing. AB - We are pleased to publish this special issue on pediatric oncology nursing. The articles in this issue provide readers with the most current clinical updates on the treatment and care of children and youth with childhood cancer. We thank Dr. Marilyn Hockenberry and the authors for their contributions to the pediatric nursing literature. PMID- 12720205 TI - The childhood leukemias. AB - Advances in treatment and prognosis of childhood leukemia are considered a remarkable success of modern medicine. Childhood leukemia, once considered a universally fatal disease, now boasts overall cure rates ranging from 75% to 85% for acute lymphocytic leukemia (ALL) and cure rates approaching 40% to 50% for acute myelogenous leukemia (AML). Inherent to this success is the expertise nurses provide when caring for children with leukemia. Understanding the classifications of leukemia and the specific therapies help direct the specialized care children with leukemia need. This article provides an overview of childhood leukemias, diagnostic and classification methods used to differentiate and evaluate childhood leukemias, and treatment strategies applied toward various forms of childhood leukemias. PMID- 12720206 TI - Solid tumors in children. AB - Solid tumors make up about 30% of all pediatric cancers. The most common types of solid tumors in children include brain tumors, neuroblastoma, rhabdomyosarcoma, Wilms' tumor, and osteosarcoma. This article reviews the etiology and treatment of these diseases, and discusses the differences between adult and pediatric solid tumors. PMID- 12720207 TI - Treatment strategies in childhood cancer. AB - Strategies for the treatment of childhood cancer have significantly evolved over time. Years ago, surgery was the only option for treating childhood cancer. Now, research has advanced treatment options to include multimodal therapy with chemotherapy, radiation, surgery and hematopoeitic stem cell transplantation. The focus on gene therapy research is also increasing. Pediatric nurses must have a thorough understanding of the treatment for childhood cancer so that as frontline healthcare providers, they give accurate information to patients and their families, deliver appropriate care, and assist with gathering data in support of ongoing research. PMID- 12720208 TI - Managing side effects of childhood cancer treatment. AB - This article focuses on the nurse's role as a member of the supportive care team for the child diagnosed with cancer and the family. The most common side effects of the cancer treatment are discussed in depth in this article. The adequate management of the side effects experienced by the child receiving cancer therapy may greatly influence the child's quality of life. PMID- 12720209 TI - Late effects of childhood cancer therapy. AB - One in every 900 young adults is a survivor of childhood cancer. Survivors may experience a wide variety of late effects stemming from the treatment they received. It is estimated that a significant portion of adult survivors of childhood cancer are not followed regularly in a center familiar with the late effects of their specific therapy. Therefore it is important for health care professionals in any setting to have an understanding of these possible late effects and encourage the survivor to seek appropriate follow-up. This article will provide a general overview of the potential late effects of childhood cancer therapy. PMID- 12720210 TI - Federally sponsored insurance programs for children: the State Children's Health Insurance Program. PMID- 12720211 TI - Sheltering children. PMID- 12720212 TI - Health hazards and risks for abuse among child labor in Jordan. AB - The phenomenon of child labor has become the main concern of developing countries. The literature related to child labor discusses the causes and socioeconomic factors contributing to child labor but few studies examined risks for physical, emotional, and sexual abuse among child labor. Therefore, the purpose of this qualitative study was to identify the characteristics of working children as well as identify health risks and risks for abuse associated with child labor. A convenience sample of 41 children working in the industrial city of Irbid were interviewed by using a semistructured interview guide. Findings revealed that children who were engaged in child labor were poor and dropped out of school to assist their families financially. The findings also showed that children were exposed to health hazards such as inhaling chemicals and physical injuries. Physical, verbal, and sexual abuse also were present among these children. The findings from this study concluded that child labor creates an unhealthy environment for socially disadvantaged children. Health professionals, including pediatric and community health care providers, have an instrumental role that needs to be focused on protecting children, enforcing policies, and implementing strategies to help socially and economically disadvantaged children. PMID- 12720213 TI - Creating a halo traction wheelchair resource manual: using the EBP approach. AB - This article describes a clinically based project that used evidence-based practice (EBP). It follows the EBP process of: (1) identifying a clinical problem and stating a clinical question that focuses the process; (2) doing a literature search for best research evidence; (3) using query techniques, such as phone calls and e-mails, to determine best clinical practice among similar institutions; and (4) drawing a practice conclusion-to accept the status quo, to instigate change of practice, or to do more research. This project was an interdisciplinary effort orchestrated by the surgical programs nurses at Boston Children's Hospital. PMID- 12720214 TI - Terrorism and/or natural disasters: are we prepared to care for America's children? PMID- 12720215 TI - Chronic graft-versus-host disease. AB - Chronic graft-versus-host disease (GVHD) remains a vexing and dangerous complication of allogeneic stem cell transplantation. Mild forms of chronic GVHD are often manageable with local or low-dose systemic immunosuppression and do not affect long-term survival. In contrast, more severe forms of chronic GVHD require intensive medical management and adversely affect survival. This report reviews current concepts of the pathogenesis, clinical risk factors, classification systems, organ manifestations, and available treatments for chronic GVHD. It also provides a comprehensive listing of the published clinical trials aimed at prevention and primary treatment of chronic GVHD. PMID- 12720216 TI - Minor histocompatibility antigen-specific cytotoxic T lymphocytes generated with dendritic cells from DLA-identical littermates. AB - Donor cytotoxic T lymphocytes (CTL) specific for minor histocompatibility antigens (mHA) mediate the graft-versus-host effect whereas host mHA-specific CTL mediate graft rejection in the setting of major histocompatibility complex identical allogeneic hematopoietic stem cell transplantation. Development of a large animal model from which mHA-specific CTL can be isolated would accelerate translation in clinical studies to improve control of the graft-versus-host effect as well as prevention of graft rejection in sensitized hosts. The aims of the current study were to isolate mHA-specific CTL from dog leukocyte antigen identical littermate nonsensitized recipients before transplantation, from stable mixed hematopoietic chimeras, and from dogs sensitized to mHA after graft rejection. Donor dendritic cells (DCs) were cultured from bone marrow-derived CD34(+) cells and were used to stimulate recipient T lymphocytes on days 1, 10, and 20 of CTL culture. We reliably generated and expanded mHA-specific CTL ex vivo from sensitized dogs that were given a donor-specific blood transfusion to boost immune recall after graft rejection after a nonmyeloablative transplantation. The mHA-specific cytotoxicity measured by (51)Cr release assay was enriched from less than 5% in the starting population of sensitized peripheral blood mononuclear cells to a median of 63% after 4 weeks in CTL culture. The expanded mHA-specific CTLs were not tissue-specific: hematopoietic cells, fibroblast, and stromal cell lines were lysed in an mHA-specific manner. Allogeneic DCs, but not peripheral blood mononuclear cells, were necessary for stimulating ex vivo expansion of mHA-specific CTL. We were unable to generate mHA specific CTL from nonsensitized dogs before transplantation, from previously sensitized dogs but without recent recall immunization, or from stable mixed hematopoietic chimeras. We conclude that after recent in vivo sensitization, large-scale ex vivo expansion of mHA-specific CTL was feasible using allogeneic DCs. PMID- 12720217 TI - Post-hematopoietic cell transplantation control of graft-versus-host disease by donor CD425 T cells to allow an effective graft-versus-leukemia response. AB - After allogeneic hematopoietic cell transplantation (HCT), the high inverse correlation between graft-versus-host disease (GVHD) and leukemic relapse requires that calculated measures be taken to reduce GVHD pathology while retaining the graft-versus-leukemia (GVL) effect. We sought to determine whether donor CD4(+)CD25(+) regulatory T cells could control ongoing GVHD, thereby providing an initial window of time in which the alloreactive anti-host response is permitted to begin, with the intent of most effectively eliminating residual leukemia cells. Prevention of lethal GVHD by infusion of donor CD4(+)CD25(+) cells early after HCT (day 2) was achieved across a major histocompatibility complex barrier in the haploidentical C3H-->(B6xC3H)F(1) model. However, in vitro expansion of donor CD4(+)CD25(+) T cells, stimulated by recipient cells in the presence of high-dose interleukin-2, was required for successful regulation. In contrast, in the major histocompatibility complex-matched, minor histocompatibility antigen-disparate, CD8-mediated B10.BR-->CBA GVHD model, lethal disease could be completely prevented by a single infusion of freshly isolated donor CD4(+)CD25(+) cells administered as late as 10 days after HCT. Of importance, this late regulatory effect required only a 3:1 ratio of effector CD8:CD4(+)CD25(+) T cells, indicating a strong potential for the delayed infusion of CD4(+)CD25(+) cells to control GVHD across minor histocompatibility antigen barriers. Furthermore, this regulation did not interfere with complete and lasting donor engraftment of the hematopoietic compartment. Of most significance, the day 10 infusion of donor CD4(+)CD25(+) cells into CBA HCT recipients that had been challenged with the MMCBA6 myeloid leukemia cell line did not block an effective GVL response, despite reducing lethal GVHD. These results suggest that donor CD4(+)CD25(+) T cells infused soon after transplantation can ameliorate the development of GVHD without sacrificing a sufficient GVL effect. PMID- 12720218 TI - Reduced-intensity transplantation with in vivo T-cell depletion and adjuvant dose escalating donor lymphocyte infusions for chemotherapy-sensitive myeloma: limited efficacy of graft-versus-tumor activity. AB - Reduced-intensity conditioning regimens allow application of allogeneic stem cell transplantation to greater numbers of patients with myeloma by reducing transplantation-related mortality. We prospectively evaluated the role of an approach incorporating in vivo T-cell depletion and subsequent adjuvant donor lymphocyte infusions (DLIs) as part of front-line therapy for chemotherapy sensitive multiple myeloma. Twenty patients with HLA-matched related (n = 12) or unrelated (n = 8) donors entered the study. None had previously undergone autologous transplantation. Acute graft-versus-host disease (GVHD) following transplantation was minimal (3 grade II and no grade III or IV). Nonrelapse mortality rate was relatively low (15%) compared with conventional myeloablative allogeneic transplantation series, although it remained significantly higher than in the autologous setting. Disease responses by 6 months posttransplantation were modest (2 in complete remission, 4 in partial remission, 2 were minimally responsive, 6 had no change, 3 had progressive disease, and 3 were not evaluable). Fourteen patients received escalating-dose DLI for residual/progressive disease. Three developed acute GVHD and 2 developed limited chronic GVHD. Seven demonstrated further disease responses, which appeared to be more common in those developing GVHD (5 of 5 versus 2 of 9; P =.02). All responses were associated with conversion from mixed to full donor T-cell chimerism. Response durations were disappointing (5 <12 months) and progression often occurred despite persisting full donor chimerism. Two-year estimated overall survival and current progression-free survival rates (intention to treat with DLI from 6 months) were 71% and 30%, respectively. The current approach incorporating T-cell depletion appears excessively immunosuppressive despite attempts to restore immune function with DLI. Dose escalation failed to allow convincing dissociation of graft-versus-myeloma from GVHD. Attempts to hasten immune reconstitution and to focus and amplify appropriate components of allogeneic T-cell responses will be required to increase complete remission rates and response durations. PMID- 12720220 TI - Hematopoietic and immunomodulatory effects of lytic CD45 monoclonal antibodies in patients with hematologic malignancy. AB - The CD45 antigen is present on all cells of the hematopoietic lineage. In some rodent models, lytic CD45 monoclonal antibodies (MAbs) induce complete marrow aplasia. In others, only transient myelolymphodepletion are observed, which are nonetheless sufficient to permit engraftment with fully allogeneic stem cells after otherwise ineffective doses of radiation. The in vivo effects of unconjugated cytolytic CD45 MAbs on myeloid and lymphoid cells in humans are unknown, so it is unclear if they could contribute in a similar way to conventional ablative or to nonmyeloablative preparative regimens used for stem cell transplantation (SCT). We therefore assessed the safety, myeloreductive activities, and lymphoreductive activities of the unconjugated rat anti-human CD45 MAbs, YTH25.4 and YTH54.12, in subjects who were to undergo SCT for advanced hematologic malignancy. The MAb pair bind to contiguous but nonoverlapping epitopes on CD45 and work synergistically to fix complement and recruit cellular lytic mechanisms. The MAbs were given in increasing doses up to 1600 microg/kg during 4 days, after which the patients began their conventional transplantation preparative regimen. The maximum tolerated dose of these MAbs, 400 microg/kg/d, produced marked reduction in circulating lymphoid and myeloid cells while largely sparing marrow progenitors. In 2 of 3 patients who had active leukemia at the time of study, the MAbs reduced the percentage of leukemic blast cells in bone marrow. Seven of 14 patients are disease free 610 to 1555 days post-SCT. The in vivo myeloreductive and lymphoreductive properties of lytic CD45 MAb in humans, therefore, closely parallel the activity seen in a murine model and, therefore, may be of similar value. PMID- 12720219 TI - Allografting after nonmyeloablative conditioning as a treatment after a failed conventional hematopoietic cell transplant. AB - Outcomes with conventional allogeneic hematopoietic cell transplantation (HCT) after failed HCT are typically poor. To reduce transplantation-related mortality (TRM), 55 patients (median age, 43 years; range, 18-69 years) who had failed conventional autologous (n = 49), allogeneic (n = 4), or syngeneic (n = 2) HCT received human leukocyte antigen-matched related (n = 31) or unrelated (n = 24) donor allografts after nonmyeloablative conditioning with 2 Gy of total body irradiation or 2 Gy of total body irradiation and 90 mg/m(2) of fludarabine. Postgrafting immunosuppression consisted of cyclosporine and mycophenolate mofetil. One rejection occurred. Thirty-three patients died a median of 127 days (range, 7-834 days) after HCT: 21 of relapse, 11 of TRM, and 1 of suicide. The TRM rate on day 100 was 11% with an estimated 1-year TRM rate of 20% (95% confidence interval [CI], 9% to 31%). The median follow-up among the 22 survivors is 368 days (range, 173-796 days). Seventeen of 22 survivors are progression free. One-year estimates of overall and progression-free survival rates are 49% (95% CI, 35% to 62%) and 28% (95% CI, 16% to 41%), respectively. Untreated disease at the time of allografting after nonmyeloablative conditioning increased the risk of death (hazard ratio = 2.4; P =.04). Although the length of follow-up is still short, it appears that encouraging outcomes can be achieved with nonmyeloablative conditioning in patients expected to have poor outcomes with conventional allografting. PMID- 12720221 TI - Re: intravenous versus oral busulfan--perhaps not as different as suggested. PMID- 12720232 TI - Brachytherapy for in-stent restenosis: the problem is one of commitment. PMID- 12720231 TI - Use of restenting should be minimized with intracoronary radiation therapy for in stent restenosis. AB - Restenting at the time of intracoronary radiation therapy (IRT) for in-stent restenosis (ISR) potentially increases the risk of late total occlusion (LTO) of the treated vessel. Prolonged antiplatelet therapy with clopidogrel (6 months) has been shown to be effective in reducing LTO risk. The purpose of this study was to assess the impact of restenting on clinical outcomes following IRT for ISR with 6 months of clopidogrel. We retrospectively evaluated 1,275 patients with 6 months clinical follow-up who were enrolled in radiation trials for ISR using gamma- and beta-emitters conducted at Washington Hospital Center. Patients were analyzed according to whether additional stents were deployed at the time of IRT. The predominant indication for restenting was to optimize the final angiographic result in the event of tissue prolapse or to cover edge dissections. All patients received a minimum of 6 months of clopidogrel. Baseline clinical and angiographic characteristics were similar between the restented and nonrestented groups. Radiation was delivered successfully in all cases. At 6 months, patients treated with additional stents and IRT had a significantly higher rate of target vessel revascularization than patients without additional stents (24.6% vs. 18.7%; P = 0.011). Restenting caused more frequent late thrombosis, late total occlusion, and Q-wave myocardial infarction than no restenting (4.0% vs. 2.2%, P = 0.09; 6.1% vs. 4.3%, P = 0.14; and 1.9% vs. 0.4%, P = 0.009, respectively). Restenting for the treatment of ISR is associated with increased adverse events and should be avoided after intracoronary radiation therapy for in-stent restenosis, as restenting results in a higher recurrence rate and the potential for increased late total occlusion. PMID- 12720233 TI - Randomized comparison of the coil-design Crossflex and the tubular NIR stent. AB - The purpose of this study was to compare the angiographic outcome of implantation of the coil-design Crossflex stent with the tubular NIR stent for treatment of coronary artery stenoses. Two hundred twenty-three patients with one genuine coronary artery lesion were randomized to implantation with a 15 mm Crossflex stent (n = 112) or a 16 mm NIR stent (n = 111). The patients had angiographic follow-up after 6 months. Primary endpoint was minimal luminal diameter (MLD) after 6 months. There was a similar clinical outcome in the two groups. At 6 month follow-up, the MLD was significantly lower in the Crossflex group (1.94 +/- 0.79 mm) than in the NIR group (2.37 +/- 0.84 mm; P < 0.001). Early gain was the same in the two groups. Late loss and percent diameter stenoses were significantly higher in the Crossflex group. The binary restenoses rate was 26% and 17% in the Crossflex and the NIR groups, respectively (P = NS). The coil design Crossflex stent was found to be inferior to the tubular NIR stent concerning late loss and MLD at 6-month follow-up. PMID- 12720234 TI - Procedural complications following diagnostic coronary angiography are related to the operator's experience and the catheter size. AB - Cardiac catheterization is performed routinely in hospitals all around the world. Extensive analysis of complications has been performed in the 1980s and early 1990s. However, because of the new therapeutic innovations based on advanced catheter technologies, these data may not apply to the present situation. Still, there are few data about procedural complications of diagnostic cardiac catheterization over the last 10 years. A total of 7,412 consecutive diagnostic cardiac catheterizations were performed between January 1990 and December 2000 and prospectively assessed in a registry. There were a total of 63 complications, of which 40 were minor and 23 major. Thus, the overall complication rate was 0.8%, with a mortality rate of 0%. Univariate analysis showed lower overall complication rate of senior physicians (> 500 coronary angiographies performed; OR = 0.58; 95% CI = 0.34-0.98; P = 0.04), smaller catheter size (< 6, 6, > 6 Fr: OR = 2.6; 95% CI = 1.53-4.41; P = 0.0004), and a higher rate in patients having left and right heart catheterization (OR = 2.62; 95% CI = 1.46-4.7; P = 0.003). Major complications were associated with larger catheters (< 6, 6, > 6 Fr: OR = 2.35; 95% CI = 1.0-5.51; P = 0.05), whereas vascular complications occurred more often with higher body weight (per 10 kg: OR = 1.4; 95% CI = 1.01-1.95; P = 0.04). Overall complication rate in diagnostic coronary angiography is very low and related to the experience of the performing cardiologist and catheter size. The only predicting risk factors for major complications in coronary angiography were catheter size and body weight. PMID- 12720235 TI - Complication-free diagnostic catheterization: In search of perfection. PMID- 12720236 TI - Impact of angina class on inhibition of platelet aggregation following clopidogrel loading in patients undergoing coronary intervention: do we need more aggressive dosing regimens in unstable angina? AB - Pretreatment with thienopyridines has been shown to improve clinical outcomes in patients undergoing percutaneous coronary intervention (PCI). We determine the impact of angina class on inhibition of platelet aggregation (IPA) following clopidogrel loading. Seventy-two patients (mean age, 64 +/- 11 years; 76% male) were pretreated with 450 mg of clopidogrel at least 3 hr prior to PCI. All patients received ASA 325 mg prior to the procedure. Patients were classified into two groups according to angina class: group 1 = stable angina or Braunwald class 1 unstable angina (UA; n = 33); group 2 = Braunwald class 2 or 3 UA (n = 39). IPA was measured prior to PCI, with the Ichor point-of-care platelet analyzer (Helena Laboratories, Beaumont, TX), using 20 microM of ADP. Group 2 patients were more likely to have prior MI (54% vs. 27%; P = 0.023), prior CABG (33% vs. 5%; P = 0.046), and received IV heparin (64% vs. 27%; P = 0.0018). Mean IPA was significantly lower in group 2 compared to group 1 (19% +/- 22% vs. 32% +/- 22%; P = 0.004). In multivariate analysis, higher angina class was independently associated with lower IPA (P = 0.018). Patients with UA undergoing PCI have a lower IPA following clopidogrel loading with 450 mg. This may indicate the possibility of clopidogrel resistance in such patients. PMID- 12720238 TI - Cardiogenic shock in acute myocardial infarction. PMID- 12720237 TI - Comparative study on transradial approach vs. transfemoral approach in primary stent implantation for patients with acute myocardial infarction: results of the test for myocardial infarction by prospective unicenter randomization for access sites (TEMPURA) trial. AB - Transradial coronary intervention (TRI) can be performed in elective patients with low incidence of access site complications. However, the feasibility of primary stent implantation by TRI is still not clear in patients with acute myocardial infarction (AMI). We prospectively randomized 149 patients out of 213 patients with AMI within 12 hr from onset into two groups: 77 patients treated by TRI (TRI group) and 72 patients by transfemoral coronary intervention (TFI; TFI group). We compared the incidences of major adverse cardiac events (MACE; repeat MI, target lesion revascularization, and cardiac death) during the initial hospitalization and 9-month follow-up periods in both groups. There were one patient who crossed over to the opposite arm, and two patients with severe bleeding complications in the TFI group. Background characteristics of patients were similar between the two groups. The success rate of reperfusion and the incidence of in-hospital MACE were similar in both groups (96.1% and 5.2% vs. 97.1% and 8.3% in TRI and TFI groups, respectively). In selected patients with AMI, primary stent implantation by TRI is feasible as compared to TFI. PMID- 12720239 TI - Percutaneous atrial septostomy for urgent palliative treatment of severe refractory cardiogenic shock due to right ventricular infarction. AB - We report a novel palliative treatment to improve refractory cardiogenic shock due to right ventricular ischemia. We performed percutaneous atrial septostomy to create an atrial septal defect, which resulted in immediate right ventricular decompression and increased left ventricular filling associated with prompt and dramatic hemodynamic and clinical improvement. PMID- 12720240 TI - Percutaneous closure of coronary pulmonary arterial fistula using catheterization laboratory trash. AB - We report a case of coronary pulmonary arterial fistula that was successfully occluded by packing the fistula with thrombogenic floppy tips of used percutaneous transluminal coronary angioplasty guidewires instead of conventional steel coils. PMID- 12720241 TI - Thermography basket catheter: in vivo measurement of the temperature of atherosclerotic plaques for detection of vulnerable plaques. AB - We have developed an intravascular thermography basket catheter to measure the temperature of the vessel wall to locate foci of inflammation. Our 3 Fr thermography basket catheter is a thermocouple-based catheter made of a nitinol expandable and externally controllable basket system loaded with nine small and flexible built-in thermosensors. It is equipped with real-time data acquisition software with a thermal resolution of 0.0001 degrees C and a sampling rate of 20 readings per second. In 10 inbred cholesterol-fed dogs with femoral (but not carotid) atherosclerosis, we found foci of warmth on the surface of atherosclerotic but not disease-free regions (P < 0.05). Marked temperature heterogeneity was also observed in the aortas of atherosclerotic Watanabe rabbits but not in normal rabbits. The catheters showed satisfactory accuracy, reproducibility, and safety. If confirmed in further studies, it has the potential to be utilized in detection of vulnerable plaques. PMID- 12720242 TI - Stent deployment for peripheral venous stenosis as a result of radiation therapy. AB - Arterial occlusive disease is a well-known complication of radiation therapy, but venous thrombosis and occlusion after radiotherapy may also occur. We report the use of an endovascular stent to treat a patient who developed peripheral venous stenosis 4 years after radiation therapy for malignant melanoma. PMID- 12720243 TI - Taking the stent in vein: interventions in the other half of the vascular tree. PMID- 12720244 TI - Endovascular closure of a large renal arteriovenous fistula. AB - A case is described of a large renal arteriovenous fistula causing long-term cardiovascular complications. Successful percutaneous embolization was achieved using temporary occlusion balloons, Guglielmi detachable coils, regular nondetachable spring coils, and cyanacrylate. The advanced therapeutic tools we used can improve the prognosis of patients with large and complicated arteriovenous fistulas and prevent surgery. PMID- 12720245 TI - Paget-Schroetter syndrome: what to do? AB - Paget-Schroetter syndrome, or primary thrombotic occlusion of the axillary subclavian vein, is diagnosed in a 42-year-old man subsequently treated with thrombolysis and anticoagulation. Treatment strategies, including early and delayed surgical options, are discussed to determine the best risk/benefit ratio. PMID- 12720246 TI - Secondary embolization of a Helex occluder implanted into a secundum atrial septal defect. AB - We report on a 2-year-old patient with atrial septal defect with a stretched diameter of 11 mm. The defect was closed uneventfully by use of a 25 mm Helex device. There was only trivial residual shunting (1 mm at the cranial margin of the device). After 36 hr, routine postinterventional echocardiography confirmed device embolization into the pulmonary artery. The child was asymptomatic. Transcatheter device retrieval with snares of different sizes, bioptomes, and retrieval forceps failed due to the mismatch of the diameter of the device and the small diameter of the pulmonary artery. Therefore, the device was retrieved surgically and the ASD closed by primary sutures. Secondary embolization of a Helex device complicated the closure of an uncomplicated atrial septal defect with a device-to-defect ratio according to the manufacturer's suggestion due to a mechanism not yet understood. PMID- 12720247 TI - Successful repositioning of an Amplatzer duct occluder immediately after inadvertent embolization in the descending aorta. AB - A case is described of the inadvertent embolization of an Amplatzer duct occluder in the descending aorta in a 10-year-old girl. It was possible to snare the device and bring it in its normal position in the aortic ductus ampulla. With a second snare, the recessed screw of the device could be caught and the device was successfully pulled back into the ductus. PMID- 12720252 TI - Risk selection and matching in performance-based contracting. AB - This paper examines selection and matching incentives of performance-based contracting (PBC) in a model of patient heterogeneity, provider horizontal differentiation and asymmetric information. Treatment effectiveness is affected by the match between a patient's illness severity and a provider's treatment intensity. Before PBC, a provider's revenue is unrelated to treatment effectiveness; therefore, providers supply treatments even if their treatment intensities do not match with the patients' severities. Under PBC, budget allocation is positively related to treatment performance; patient-provider mismatch is reduced because patients are referred more often. Using data from the state of Maine, we show that PBC leads to more referrals and better match between illness severity and treatment intensity. Moreover, we find that PBC has a positive but insignificant effect on dumping. PMID- 12720253 TI - A critique of the World Health Organisation's evaluation of health system performance. AB - The World Health Organisation's (WHO) approach to the measurement of health system efficiency is briefly described. Four arguments are then presented. First, equity of finance should not be a criterion for the evaluation of a health system and, more generally, the same objectives and importance weights should not be imposed upon all countries. Secondly, the numerical value of the importance weights do not reflect their true importance in the country rankings. Thirdly, the model for combining the different objectives into a single index of system performance is problematical and alternative models are shown to alter system rankings. The WHO statistical analysis is replicated and used to support the fourth argument which is that, contrary to the author's assertion, their methods cannot separate true inefficiency from random error. The procedure is also subject to omitted variable bias. The econometric model for all countries has very poor predictive power for the subset of OECD countries and it is outperformed by two simpler algorithms. Country rankings based upon the model are correspondingly unreliable. It is concluded that, despite these problems, the study is a landmark in the evolution of system evaluation, but one which requires significant revision. PMID- 12720254 TI - Determinants of health care utilization by German sickness fund members--with application to risk adjustment. AB - In many countries, social health insurance systems are being reformed in favor of more competition among insurers, while premiums are community rated by regulation. The implicit incentives for insurers to engage in risk selection can only be curtailed using appropriate systems of risk-adjusted equalization payments among insurers. To develop these systems, predictors of individual utilization patterns have to be identified, e.g. via regression analysis using previous utilization data. In some countries such as Germany, such data are hardly ever available. In the early nineties, a number of sickness funds participated in an experiment in which individual utilization data were collected. Our data set covers more than 70,000 members of company sickness funds over a 5-year period. We analyze socio-demographic determinants of utilization which could be used as risk adjusters in a risk equalization scheme. Our results suggest that besides age and sex, the set of risk adjusters should include income, family status and a dummy for the last year of life. PMID- 12720255 TI - Missing... presumed at random: cost-analysis of incomplete data. AB - When collecting patient-level resource use data for statistical analysis, for some patients and in some categories of resource use, the required count will not be observed. Although this problem must arise in most reported economic evaluations containing patient-level data, it is rare for authors to detail how the problem was overcome. Statistical packages may default to handling missing data through a so-called 'complete case analysis', while some recent cost analyses have appeared to favour an 'available case' approach. Both of these methods are problematic: complete case analysis is inefficient and is likely to be biased; available case analysis, by employing different numbers of observations for each resource use item, generates severe problems for standard statistical inference. Instead we explore imputation methods for generating 'replacement' values for missing data that will permit complete case analysis using the whole data set and we illustrate these methods using two data sets that had incomplete resource use information. PMID- 12720256 TI - Threats to the estimation of benefit: are preference elicitation methods accurate? AB - Stated preference methods are used to estimate the value that people place on health care. The data that emerges from these studies is used to guide health policy. However, relatively little is known about how individuals make decisions in a preference elicitation task. Two methods (willingness to pay and conjoint analysis) are considered within the context of the literature from psychology (and also environmental economics) regarding how people construct preferences, process information, and make decisions. There is substantial evidence that individuals employ heuristics (cognitive shortcuts) in order to simplify tasks they are presented with. The use of heuristics implies that people ignore much of the information they are presented with and make decisions which would not be considered rational in the economic sense. These stated preference methods assume that individuals trade between the different attributes of a good or service when making decisions - an assumption that other theories predict is wrong. The implications of this are discussed. PMID- 12720257 TI - A bibliometric analysis of health economics articles in the economics literature: 1991-2000. AB - This paper describes and analyzes trends in health economics articles indexed in the economics literature from 1991 to 2000, demonstrating the robust state of publication in the field during the past decade. While articles, pages, and the number of journals increased, single authorship declined dramatically from almost half of articles published to only one-third, and papers with four or more authors increased three-fold. Over three-fourths of articles were analyses of healthcare markets or health production, while policy oriented articles constituted the third largest share. Author concentration ratios decreased almost by half and the Herfindhal-Hirschman index of author concentration declined from 14 in 1991 to only 4 in 1999. PMID- 12720258 TI - Measles vaccination improves the equity of health outcomes: evidence from Bangladesh. AB - OBJECTIVES: This paper asks whether measles vaccination can reduce socioeconomic differentials in under five mortality rates (U5MR) in a setting characterized by extreme poverty and high levels of childhood mortality. DESIGN: Longitudinal cohort study based on quasi experimental design. SETTING: Data come from the phased introduction of a measles vaccine intervention in Matlab, Bangladesh in 1982. SUBJECTS: There were 16 270 Bangladeshi children aged 9-60 months. INTERVENTION: The intervention cohort received measles vaccine. MAIN OUTCOME MEASURES: Socioeconomic differentials in U5MR between the lowest and highest socioeconomic status (SES) quintiles in a cohort of 8135 vaccinated children and a cohort of unvaccinated age matched controls. Mantel-Haenszel rate ratios for the lowest to highest SES quintile were computed. SES was measured by factor analysis of maternal schooling, land holdings, dwelling size, and number of rooms. RESULTS: The U5MR ratio of lowest SES to highest was 2.27 (95% CI=1.62 3.19) in the unvaccinated population and 1.42 (95%CI=0.94-2.15) in the vaccinated population. The difference between unvaccinated and vaccinated U5MR ratios was statistically significant (p<0.10) and robust across alternative measures of SES. CONCLUSION: Children from the poorest quintile were more than twice as likely to die as those from the least quintile in the absence of measles vaccination. Universal distribution of measles vaccination largely nullified SES related mortality differentials within a high mortality population of children. PMID- 12720259 TI - Stratified cost-effectiveness analysis: a framework for establishing efficient limited use criteria. AB - The cost-effectiveness of new health care technologies is conditional upon who receives what therapy and under what circumstances. Understanding this heterogeneity in cost-effectiveness, health care payers often limit reimbursement of therapies to a more restrictive sub-group of patients than that indicated in a product's licensing. Such limits may be based upon clinical or demographic criteria that are prognostic of costs, outcomes or both. However, there is little guidance on how to estimate and interpret stratified cost-effectiveness analysis. In this paper we present a framework for estimating the benefits from stratification that permits consideration of both the opportunity cost resulting from a lack of adherence with criteria and the efficiency loss associated with incorporating equity concerns. PMID- 12720260 TI - Functional tumor imaging with dynamic contrast-enhanced magnetic resonance imaging. AB - Dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) is the acquisition of serial MRI images before, during, and after the administration of an MR contrast agent. Unlike conventional enhanced MRI, which simply provides a snapshot of enhancement at one point in time, DCE-MRI permits a fuller depiction of the wash-in and wash-out contrast kinetics within tumors, and thus provides insight into the nature of the bulk tissue properties. Such data is readily amenable to two-compartment pharmacokinetic modeling from which parameters based on the rates of exchange between the compartments can be generated. These parameters can be used to generate color-encoded images that aid in the visual assessment of tumors. DCE-MRI is used currently to characterize masses, stage tumors, and noninvasively monitor therapy. While DCE-MRI is in clinical use, there are also a number of limitations, including overlap between malignant and benign inflammatory tissue, failure to resolve microscopic disease, and the inconsistent predictive value of enhancement pattern with regard to clinical outcome. Current research focuses on improving understanding of the meaning of DCE-MRI at a molecular level, evaluating macromolecular and targeted contrast agents, and combining DCE-MRI with other physiologic imaging techniques such as positron emission tomography. Efforts to standardize DCE-MRI acquisition, analysis, and reporting methods will allow wider dissemination of this useful functional imaging technique. PMID- 12720261 TI - The diagnostic utility of FLAIR imaging in clinically verified amyotrophic lateral sclerosis. AB - PURPOSE: To explore the overall diagnostic ability of magnetic resonance (MR) fluid-attenuated inversion recovery (FLAIR) imaging for clinically verified amyotrophic lateral sclerosis (ALS). MATERIALS AND METHODS: Abnormal signal intensity in FLAIR images of 18 patients with ALS and 18 age-matched normal controls were scored and compared. Six patients had serial MR imaging scans within 97 days. Mann Whitney U statistics and ANOVA were used for statistical analysis. RESULTS: Hyperintensity in the subcortical white matter (SWM) and the dark line along the posterior rim of the precentral gyri were found more frequently in patients with ALS. The scores for these two signs were significantly different from those of normal controls. Hyperintensity in the corticospinal tract (CST) was found in both ALS and normal controls, but the difference was not statistically significant. ANOVA on the serial FLAIR studies revealed no significant difference in the scores among the series. The hyperintensity in SWM had a sensitivity of 56% and specificity of 94%, while the dark line in the motor cortex had a 74% sensitivity and 67% specificity. CONCLUSION: Hyperintensity in SWM and the dark line along the posterior rim of the precentral gyri appeared more frequently in the patients with ALS. SWM hyperintensity has a better specificity in association with clinically verified ALS, while the motor dark line has a better sensitivity. No correlation was found between the FLAIR findings and progression of the disease. PMID- 12720262 TI - Feedback-assisted three-dimensional reconstruction of the left ventricle with MRI. AB - PURPOSE: To develop and test a new technique for rapid, accurate three dimensional (3D) reconstruction of the left ventricle (LV) and calculation of its volume parameters, with images from multiple orientations and interactive feedback. MATERIALS AND METHODS: The ventricular surface was fit to a number of user-placed guide points in magnetic resonance (MR) images using bivariate smoothing splines. A 3D model was reconstructed and the LV volumes were calculated at both end diastole (ED) and end systole (ES). This technique was validated using a phantom, and applied to studies of 18 patients and four volunteers (N = 22) imaged on a 1.5-T clinical scanner. The results of the 3D method were compared to the standard 2D short-axis slice summation technique, which is widely used for the analysis of cardiac function. RESULTS: There was excellent agreement between the computed volume of the phantom using the 3D modeling method and the actual volume (190.50 mL +/- 3.06 mL, and 191.0 mL +/- 2.5 mL, respectively). There was good correlation between the volumes calculated with our 3D model and the slice summation technique (ED volume (EDV) difference, 6.36% +/- 8.99% [mean +/- SD]; ES volume (ESV), 0.92% +/- 14.75%; stroke volume (SV), 10.54% +/- 13.95%; ejection fraction (EF), 4.22% +/- 9.16%). The 3D method was found to be more accurate than the slice summation technique for calculating LV volumes and mass from images of different slice orientations. Variations in the parameters between the two separate orientations using the 3D model vs. the slice summation method were as follows: EDV: 2.11% +/- 1.52% vs. 10.36% +/- 9.33%; ES volume: 2.76% +/- 1.64% vs. 6.39% +/- 3.62%; SV, 3.02% +/- 4.38% vs. 18.84% +/- 15.30%; EF, 2.03% +/- 2.16% vs. 8.58% +/- 6.73%; and LV mass: 4.77% +/ 2.41% vs. 24.59% +/- 6.41%. Differences in the ES volume due to the inclusion or exclusion of the most basal slice were found to be lower with the 3D model (6.90% +/- 3.83%) compared to the slice summation method (25.04% +/- 6.15%). CONCLUSION: 3D models can be used to accurately determine ventricular volume parameters. Results can be obtained using images from a variety of orientations, providing greater flexibility during image acquisition and possibly reducing the number of images needed for analysis. Feedback is provided to assist the analysis by providing a continuous update of the LV shape and volume. This feature allows the user to determine LV parameters to a predefined accuracy or to terminate the analysis when the parameters are not changing. This method is not restricted to multislice cine imaging in a single or prescribed slice orientation, and can be used for quick, accurate, and interactive analysis of cardiac function. PMID- 12720263 TI - The impact of spatial resolution and respiratory motion on MR imaging of atherosclerotic plaque. AB - PURPOSE: To examine the impact of spatial resolution and respiratory motion on the ability to accurately measure atherosclerotic plaque burden and to visually identify atherosclerotic plaque composition. MATERIALS AND METHODS: Numerical simulations of the Bloch equations and vessel wall phantom studies were performed for different spatial resolutions by incrementally increasing the field of view. In addition, respiratory motion was simulated based on a measured physiologic breathing pattern. RESULTS: While a spatial resolution of > or = 6 pixels across the wall does not result in significant errors, a resolution of < or = 4 pixels across the wall leads to an overestimation of > 20%. Using a double-inversion T2 weighted turbo spin echo sequence, a resolution of 1 pixel across equally thick tissue layers (fibrous cap, lipid, smooth muscle) and a respiratory motion correction precision (gating window) of three times the thickness of the tissue layer allow for characterization of the different coronary wall components. CONCLUSIONS: We found that measurements in low-resolution black blood images tend to overestimate vessel wall area and underestimate lumen area. PMID- 12720264 TI - Atherosclerotic disease distribution in carotid and vertebrobasilar arteries: clinical experience in 100 patients undergoing fluoro-triggered 3D Gd-MRA. AB - PURPOSE: To determine how often three-dimensional gadolinium-enhanced magnetic resonance angiography (3D Gd-MRA) shows disease outside of the standard two dimensional time-of-flight magnetic resonance angiography (2D TOF-MRA) imaging volume. MATERIALS AND METHODS: One hundred consecutive patients with known or suspected cerebrovascular disease were examined using fluoro-triggered 3D Gd-MRA with a 1.5 Tesla scanner. All vessel segments from the aortic arch to the circle of Willis were independently evaluated for the presence of occlusive disease, ulceration, aneurysm, and anatomic variations by two radiologists blinded to clinical information. Branch vessel and circle of Willis visibility were also assessed. Kappa statistics were calculated to determine interobserver agreement. RESULTS: Interobserver agreement for the degree of stenosis was good to excellent (kappa = 0.83-1.0). Disagreement was more frequent in the cases of mild stenoses and was less frequent for stenoses > or = 50%. In 38% of the patients, 3D Gd-MRA demonstrated either ulcerated plaques in carotid arteries (N = 14) or stenosis > or = 50% in great vessel (N = 20) and/or vertebral artery (N = 38) origins, none of which can be readily evaluated on standard TOF-MRA. CONCLUSION: Fluoro triggered 3D Gd-MRA provides a comprehensive evaluation of the carotid arteries including all vessel segments from the aortic arch to the circle of Willis. PMID- 12720265 TI - Time-resolved contrast-enhanced magnetic resonance angiography in pediatric patients using sensitivity encoding. AB - PURPOSE: To evaluate the role of time-resolved contrast-enhanced magnetic resonance angiography (CE-MRA) using sensitivity encoding in imaging the thoraco abdominal vessels in pediatric patients. MATERIALS AND METHODS: Thoraco-abdominal vessels of 22 pediatric patients (median age = 5 years) were evaluated with a 3D CE-MRA technique in combination with SENSE following a 0.2 mmol/kg injection of Gd-chelate. The acquisition parameters were as follows: TR/TE = 5/1.1 msec; flip angle = 40 degrees; in-plane phase encoding steps were reduced by a factor of 2 using sensitivity encoding (SENSE); 3D volume acquisition was repeated four to eight times consecutively during free breathing (four to eight dynamics) with a mean temporal resolution of 6.8 seconds/dynamic; and mean acquired voxel size = 1.4 x 1.7 x 3.1 mm (reconstructed as 1.4 x 1.4 x 1.55 mm). Arterial-to-venous signal intensity ratios (AVRs) were computed for each dynamic. RESULTS: All images were successfully reconstructed and were of diagnostic quality. The AVRs of prepeak, peak, and postpeak arterial volumes were 1.0 +/- 0.5, 6.1 +/- 3.3, and 1.3 +/-0.9, respectively, indicating good arterial-to-venous separation. The signal-to-noise ratio (SNR) of the peak arterial volume was 41 +/- 26. CONCLUSION: Our results suggest that it is feasible to apply SENSE to a conventional 3D CE-MRA technique in a time-resolved fashion for imaging the thoraco-abdominal vessels in pediatric patients during free breathing. PMID- 12720266 TI - Using a 1 M Gd-chelate (gadobutrol) for total-body three-dimensional MR angiography: preliminary experience. AB - PURPOSE: To determine whether higher concentrated gadolinium chelates are advantageous for the recently introduced concept of total-body magnetic resonance angiography (MRA), allowing whole-body coverage, extending from the carotid arteries to the runoff vessels, in merely 72 seconds. MATERIALS AND METHODS: Total-body three-dimensional (3D) MRA using a 1 M Gd-chelate (gadobutrol, Gadovist, Schering, Berlin, Germany) at a dosage of 0.2 mmol/kg body-weight (biphasic injection protocol: 1.3 mL/second and 0.7 mL/second) was performed on three healthy volunteers and ten consecutive patients with DSA-documented peripheral vascular disease. Separated by at least 72 hours, the three healthy volunteers also underwent the same MRA-protocol, using gadopentetate dimeglumine in equimolar dosages. RESULTS: Compared to equimolar dosages, mean signal-to noise ratio (SNR) and contrast-to-noise ratio (CNR) values in the three volunteers were significantly higher (up to 32.5% for the arteries of the thighs and calves) using gadobutrol. In the ten patients, gadobutrol-based total-body MRA accurately assessed significant stenoses (luminal narrowing > 50%) with sensitivities and specificities of 96.2% (95% CI 0.83-0.97) and 95.7% (95% CI 0.84-0.96), respectively, compared to digital subtraction angiography. CONCLUSION: The MRA image quality for total-body MRA provided by the administration of gadobutrol is superior to that obtained following administration of an identical dose of gadopentetate dimeglumine, and therefore shows promise for use as a comprehensive single exam assessing the entire arterial system for the presence of atherosclerotic disease manifestations. PMID- 12720267 TI - Volume-selective 3D turbo spin echo imaging for vascular wall imaging and distensibility measurement. AB - PURPOSE: To use a volume-selective 3D turbo spin echo (TSE) technique to image the carotid artery wall and measure distensibility. MATERIALS AND METHODS: A high resolution volume-selective 3D TSE sequence has been developed. Volume selection is accomplished by orthogonal gradients for the 90 degrees and 180 degrees excitations and allows a 3D volume of vessel wall to be imaged in a relatively short time. The technique has been developed to allow imaging at any defined phase of the cardiac cycle so that the vascular function and distensibility can be studied. RESULTS: Scan efficiency is increased by the reduced phase encode field of view (FOV) (k(y) steps) by the use of selective volume excitation. Significantly more slices (k(z) phase encode steps) for vessel coverage can be acquired with the same scan time as that of the conventional sequence while maintaining signal-to-noise ratio (SNR) levels. The practical value of the technique was demonstrated on 10 normal subjects with high-resolution vessel distensibility measurements of the carotid arteries. CONCLUSION: A volume selective TSE method has been used for carotid artery wall imaging and measurement of distensibility in normal subjects. Larger coverage of the vessel, and therefore more information for clinical diagnostics, was achieved with the same overall scan time with an SNR comparable to that of 2D full FOV images. PMID- 12720268 TI - Rapid hyperpolarized 3He diffusion MRI of healthy and emphysematous human lungs using an optimized interleaved-spiral pulse sequence. AB - PURPOSE: To develop and validate an interleaved-spiral diffusion pulse sequence capable of hyperpolarized (3)He MR imaging of the whole lung in less than 10 seconds. MATERIALS AND METHODS: Hyperpolarized (3)He diffusion measurements were performed in seven healthy volunteers and five patients with emphysema using an interleaved-spiral pulse sequence that provided 11 contiguous 15-mm thick coronal ADC maps, with an in-plane resolution of 3.9 mm, covering the whole lung in 5.5 seconds. The resulting means and SDs of ADC values were compared statistically to those from a gradient-echo pulse sequence with identical resolution and diffusion weighting gradients that acquired five ADC maps in 10.5 seconds. RESULTS: High quality diffusion-weighted interleaved-spiral images covering the whole lung were obtained, and showed no significant susceptibility-induced image degradation compared to corresponding gradient-echo images. On a subject-by-subject basis, the means and SDs of ADC values for the interleaved-spiral technique were not statistically different from those for the gradient-echo technique. The mean ADC values from the two techniques were highly correlated on a section-by-section basis (R = 0.99). CONCLUSION: The interleaved-spiral diffusion pulse sequence permits rapid acquisition of contiguous ADC maps covering the whole lung during a short breath-hold period, and provides ADC values that are statistically equivalent to those from standard gradient-echo techniques. PMID- 12720269 TI - Comparison between signal-to-noise ratio, liver-to-muscle ratio, and 1/T2 for the noninvasive assessment of liver iron content by MRI. AB - PURPOSE: To compare different MRI-derived parameters, i.e., liver signal-to-noise ratio (LSNR), liver-to-muscle ratio (LMR) and liver transversal relaxation rate (R2), in terms of their correlation with the ex vivo determined iron content in an experimental model of liver iron overload. MATERIALS AND METHODS: Multi-echo spin echo (SE) images of the liver were acquired at 4.7 T from a group of 33 male wistar rats subjected to a high iron content diet for feeding periods ranging from 2 to 50 days. Liver transversal relaxation time, liver signal-to-noise ratio, and liver-to-muscle ratio were measured over the same region of interest in order to get a direct comparison between these parameters. After MRI experiments, the rats were sacrificed and the liver iron content was measured ex vivo by atomic absorption spectroscopy. RESULTS: The iron content is better correlated to the LSNR than to the other parameters (LMR, R2). CONCLUSION: The finding that liver signal-to-noise ratio is better correlated to the iron content than the liver T2 relaxation rate is relevant for clinical applications of MRI because a T2 determination is more time-consuming, both for acquisition and postprocessing of images, than a simple SNR determination. PMID- 12720270 TI - MRI of focal nodular hyperplasia (FNH) with gadobenate dimeglumine (Gd-BOPTA) and SPIO (ferumoxides): an intra-individual comparison. AB - PURPOSE: To compare the efficacy of two different MR contrast agents for the detection and diagnosis of focal nodular hyperplasia (FNH). MATERIALS AND METHODS: Fifty patients with 83 FNH lesions detected on spiral CT were studied in two different MRI sessions with Gd-BOPTA (MultiHance) and ferumoxides (Endorem). MRI with Gd-BOPTA was performed precontrast (T1wGRE and T2wTSE sequences) and during the dynamic and late (1-3 hours) phases after injection (T1wGRE sequences only). MRI with ferumoxides (T1wGRE and T2wTSE sequences) was performed before and at least 30 minutes after injection. Hyper- or isointensity of FNH in the late phase was considered typical for Gd-BOPTA, while isointensity or lesion hypointensity was considered typical for ferumoxides. RESULTS: With Gd-BOPTA, 83 FNH lesions (100%) appeared hyperintense during the arterial phase of dynamic MRI. All but one lesion was iso- or slightly hyperintense in the portal-venous and equilibrium phases. In the late phase, 81 FNH lesions were hyper- or isointense to the surrounding parenchyma, with two lesions appearing slightly hypointense. With ferumoxides, a significant (P < 0.001) number (21/83, 25.3%) of FNH lesions (mean diameter = 16.8 +/- 6.6 mm) were not visible. Of the visible FNH lesions, 38/62 were slightly hyperintense, and 24/62 were isointense to the surrounding parenchyma on the T2wTSE images. On the T1wGRE images, 42/62 lesions were isointense, 19/62 were slightly hyperintense, and one lesion was slightly hypointense. Seventeen lesions in 12 patients with previous neoplasia were all detected after Gd-BOPTA administration, whereas only nine of these 17 lesions (52.9%) were detected after ferumoxide administration. Two of these nine lesions showed atypical enhancement features. CONCLUSION: Gd-BOPTA-enhanced MRI is significantly better than ferumoxide-enhanced MRI for the identification and characterization of FNH. PMID- 12720271 TI - Ex vivo magnetic resonance microscopy of an osteochondral transfer. AB - A 49-year-old woman with right knee pain and a chondral defect on the medial femoral condyle underwent an osteochondral transfer. The patient initially had pain relief, but then sustained a twisting injury and had progressive chondromalacia and pain on the affected side. She subsequently underwent a total knee replacement, and the tissue from the osteochondral transfer (OATS) site was harvested for analysis. In vitro MR microimaging of the excised joint segment revealed undamaged, full-thickness cartilage on the OATS plug, intact cartilage on the posterior condyle, and severely thinned and damaged cartilage on the anterior condyle. Alcian blue-stained sections revealed that proteoglycans were present throughout the OATS core but were nearly absent in the native cartilage. Quantitative T(1) data acquired after equilibration with Gd-DTPA indicated a distribution of matrix fixed charge in the OATS plug and anterior tissue that agreed well with histology and literature observations, while the posterior native cartilage appeared to have fixed charge similar to that of the OATS tissue. Histology revealed poor graft integration between OATS and native cartilage, with a distinct layer of fibrous tissue at the posterior interface. MRI images, by comparison, showed a hypointense feature at the posterior interface but uniform intensity across the anterior interface. Quantitative T(2), magnetization transfer and T(1) data acquired with and without gadolinium contrast showed dependences on depth, location, and pathology that were consistent with measurements reported in the literature for articular cartilage. PMID- 12720273 TI - Initial experiences with in vivo intravascular coronary vessel wall imaging. AB - PURPOSE: To evaluate potential use of a loopless internal receiver coil for in vivo coronary artery vessel wall imaging in five domestic swine. MATERIALS AND METHODS: Intravascular free-breathing black blood coronary vessel wall imaging was performed using a previously described double inversion fast spin echo technique after x-ray guided placement of an internal receiver coil in or in close proximity to the target vessel (LAD, LCX). RESULTS: Image quality using the phased array coil was reproducible, while image quality with the internal receiver coil was heavily dependent on coil position with respect to the examined artery, and likely also dependent on blood flow and/or cardiac-related coil motion. With internal coil placement in the left circumflex coronary artery, images of the left anterior descending vessel wall appeared similar or superior compared to commercially available phased array surface coil images. With coil placement in the target vessel itself, imaging was suboptimal because of the extremely high signal intensity (hotspot) in close proximity to the vessel wall, leading to low contrast between the vessel wall and the surrounding tissues and blood. CONCLUSION: In this study, we demonstrate the feasibility of in vivo intravascular coronary vessel wall imaging. Continued research is necessary to minimize coil motion and optimize coil sensitivity algorithms. PMID- 12720272 TI - Contrast-enhanced peripheral MR angiography at 3.0 Tesla: initial experience with a whole-body scanner in healthy volunteers. AB - PURPOSE: To report preliminary experience with contrast-enhanced magnetic resonance angiography (CE-MRA) of the peripheral arteries on a 3.0 T whole-body scanner equipped with a prototype body coil. MATERIALS AND METHODS: Four healthy volunteers were imaged on the 3.0 T system and, for comparative purposes, two of the subjects were also imaged on a commercially available 1.5 T whole-body system. To investigate field strength influence on objective image quality, signal-to-noise (SN) and contrast-to-noise (CN) ratios were calculated for named vessels from the infrarenal aorta to the ankles at both field strengths. Comparable imaging protocols were used at both field strengths. In addition, two reviewers, blinded for field strength, gave subjective image quality scores (three-point scale). RESULTS: SN and CN ratios were approximately equal on both systems (variation < or =9%) for the iliac and proximal upper leg stations. For the popliteal and lower leg stations SN ratios were 36% and 97% higher, and CN ratios were 44% and 127% higher, at 3.0 T. Subjective image quality at 3.0 T was substantially better for the distal upper and lower legs. CONCLUSION: Contrast enhanced peripheral MRA is possible at 3.0 T when an imaging protocol similar to a current state-of-the-art 1.5 T protocol is used. Objective and subjective image quality at 3.0 T is comparable for the iliac and upper legs but better for the popliteal and lower leg arteries. PMID- 12720274 TI - Pair of resonant fiducial markers for localization of endovascular catheters at all catheter orientations. AB - PURPOSE: To test wireless resonance circuits (RC) to be used as fiducial marker of endovascular catheters during MR-guided interventions. Current markers loose their signal enhancement for certain catheter orientations. The purpose of this study was to test a marker setup which overcomes this orientation problem. MATERIALS AND METHODS: The markers were constructed from a pair of two RCs. The RCs were individually tuned and the coil axes were oriented perpendicular to each other in order to decouple the two RCs. The markers were mounted on the tip of endovascular catheters and tested in vitro and in one porcine in vivo experiment. RESULTS: An intense MR signal at similar signal levels was noted at all catheter orientations. In the in vivo experiment the markers allowed for fast and reliable MR guidance of the catheters. CONCLUSION: A pair of two individually tuned and decoupled RCs is well suited for MR guidance of endovascular catheters. PMID- 12720275 TI - Multiple-point adsorption of terbium ions by lead ion templated thermosensitive gel: elucidating recognition of conformation in gel by terbium probe. AB - Lead ion templated thermosensitive heteropolymer gel which has recognition ability of methacrylate pairs has been synthesized and characterized. The gel consists of a main monomer component, N-isopropylacrylamide (NIPA), responsible for volume phase transition, methacrylic acid (MAA) moieties imprinted as pairs to adsorb terbium ions and cross-links. An imprinting technique was applied using lead ion complex with methacrylate ligands in dioxane media. After gel was obtained, lead ions were removed by washing and the imprinted gel showed strong binding ability to terbium ions, comparable with that of the non-imprinted gel prepared without lead ions. It was found that the Tb(3+) fluorescence intensity was considerably increased upon binding this ion to both imprinted and non imprinted gels, but the largest enhancement of fluorescence intensity was observed when Tb(3+) was bound to imprinted gel in shrunken state. This is because of the decrease of coordinated water molecules on Tb(3+) and the strong binding of this ion to methacrylate pairs which are encoded within the weakly cross-linked network of imprinted gel. PMID- 12720276 TI - X-ray structural analysis of the ligand-recognition mechanism in the dual affinity labeling of c-type lysozyme with 2',3'-epoxypropyl beta-glycoside of N acetyllactosamine. AB - In spite of the belonging to the same c-type lysozyme family, hen egg-white lysozyme (HEWL) was much less susceptible to the dual-affinity labeling with 2',3'-epoxypropyl beta-glycoside of N-acetyllactosamine (Galbeta1,4GlcNAc-Epo) than human lysozyme (HL). The three-dimensional structures of the HEWL labeled with single Galbeta1,4GlcNAc-Epo and the Glu102-mutant HL labeled with double Galbeta1,4GlcNAc-Epo were determined by X-ray crystallography at resolutions of 1.85 and 2.0 A, respectively. The overall conformation and the interaction mode of the carbohydrate ligand part in the singly labeled HEWL and the doubly labeled Glu102-mutant HL were basically identical to those of the correspondingly labeled wild-type HL with minor alterations in some stereochemical parameters. A detailed comparison of the structures revealed the key protein-carbohydrate and carbohydrate-carbohydrate interactions essential for the dual labeling. It was suggested that the difference in the efficiency of the dual labeling was caused by the structural difference between Gln104 in HL and Asn103 in HEWL. The relevance to our previous study and the carbohydrate-carbohydrate interaction on cell-surface membranes were discussed. PMID- 12720277 TI - Comparison of the three-dimensional structures of a human Bence-Jones dimer crystallized on Earth and aboard US Space Shuttle Mission STS-95. AB - Crystals of a human (Sea) Bence-Jones dimer were produced in a capillary by vapor diffusion under microgravity conditions in the 9 day US Space Shuttle Mission STS 95. In comparison to ground-based experiments, nucleation was facile and spontaneous in space. Appearance of a very large (8 x 1.6 x 1.0 mm) crystal in a short time period is a strong endorsement for the use of microgravity to produce crystals sufficiently large for neutron diffraction studies. The Sea dimer crystallized in the orthorhombic space group P2(1)2(1)2(1), with a = 48.9 A, b = 85.2 A, and c = 114.0 A. The crystals grown in microgravity exhibited significantly lower mosaicities than those of ground-based crystals and the X-ray diffraction data had a lower overall B factor. Three-dimensional structures determined by X-ray analysis at two temperatures (100 and 293 K) were indistinguishable from those obtained from ground-based crystals. However, both the crystallographic R factor and the free R factor were slightly lower in the models derived from crystals produced in microgravity. The major difference between the two crystal growth systems is a lack of convection and sedimentation in a microgravity environment. This environment resulted in the growth of much larger, higher-quality crystals of the Sea Bence-Jones protein. Structurally, heretofore unrecognized grooves on the external surfaces of the Sea and other immunoglobulin-derived fragments are regular features and may offer supplementary binding regions for super antigens and other elongated ligands in the bloodstream and perivascular tissues. PMID- 12720278 TI - Therapeutic approaches to superantigen-based diseases: a review. AB - Superantigens secreted by the bacterial pathogen Staphyloccocus aureus are extremely potent toxins that overstimulate the host immune system by binding to the MHC class II and T cell receptors and activating a large population of T cells. Superantigen infection has been shown to be the causative agents in acute diseases, food poisoning and toxic shock syndrome, and in more chronic conditions such as inflammatory skin diseases. In addition to the toll on public health, S. aureus superantigens also represent a potential biothreat to our national security. To address these risks, a number of different therapeutic strategies have been developed that target different aspects of the pathogenic mechanism of S. aureus and superantigen infection. These therapies, which encompass strategies as diverse as production of neutralizing antibodies, inhibitory peptide/receptor design and blockage of superantigen gene transcription, are being tested for treatment of established S. aureus infections in pre- and post-exposure scenarios. In this review, we will describe these different strategies and their efficacies in inhibition of superantigen-induced effects in the host, and present the future outlook for successfully producing therapies for superantigen-based disease. PMID- 12720279 TI - Recognition promoted by Zn2+ between phenanthroline bridging polyaza ligands and nucleotides--Zn2+ acts as 'messenger' between the receptor and substrate. AB - The stability constants of the supramolecular complexes formed between L ((a,b,c,d)) or their Zn(2+) complexes, and adenosine 5'-triphosphate (ATP) in aqueous solution were determined by potentiometric titrations (25 degrees C, I = 0.1 mol dm(-3) KNO(3)). The results show that protonated aliphatic-substituted L (a,d) and aromatic-substituted L (b,c) ligands and/or Zn(II) ion can efficiently recognition the substrate, ATP. All of the equilibrium studies, (1)H and (31)P nuclear magnetic resonance spectra indicate that multiple interactions, including coordination, pi-stacking, ion-pairing, H-bonding, and possible ion-pi-donor, hydrophobic and even van der Waals interactions exist in the Zn(II)-L-ATP systems. On the other hand, the recognition of the substrates by the protonated ligands was significantly promoted by the addition of Zn(II), which leads to coordination competition between the mixed ligands, L and nucleotide. In Zn(II)/L/ATP systems the tendency for phosphate chain to receive proton and metal ion increases, facilitating the cleavage of the phosphate chain of the nucleotide. PMID- 12720280 TI - On the 17O correction for CO2 mass spectrometric isotopic analysis. AB - To calculate delta(13)C from raw CO(2) isotope data, the ion beam ratio of m/z 45 to 44 is corrected for the contribution arising from the contribution of (17)O bearing molecules. First, a review on the current state of (17)O-corrections for CO(2) mass spectrometry is presented. The three correction algorithms that are generally in use, however, do produce biased delta(13)C values, and the bias is actually larger than the precision of modern isotope ratio mass spectrometers. The origin of this bias is twofold: different values for (17)R(VPDB-CO2) as well as different values for lambda are used in the correction algorithms. Despite both values being of high importance, large discrepancies between the absolute values published for (17)R(VPDB-CO2) appear to be the main reason for the delta(13)C biases. Next, the question of how to choose the value of lambda to best be used is considered. Natural (e.g. tropospheric) CO(2) as well as primary reference materials (PDB and NBS-19), having been in isotope exchange with water, are assumed to lie on the fractionation line for waters. On this ground, lambda = 0.5281 +/- 0.0015, as determined for waters (Meijer and Li, Isot. Environ. Health Stud., 1998; 34: 349-369), is suggested to be a base for the (17)O-correction algorithm. Finally, an approach to determine the absolute value for (17)R(VPDB CO2), based on data of relative isotope measurements on two CO(2) gases having a large (17)O difference, is discussed and algebraic formulas are considered. Experimental data and new numerical values determined for (17)R(VPDB-CO2) and (17)R(VSMOW) are given in a companion paper. PMID- 12720281 TI - A redetermination of absolute values for 17RVPDB-CO2 and 17RVSMOW. AB - In a companion paper in this issue we presented a review of the current state of (17)O-corrections for CO(2) mass spectrometry and considered an approach (including algebraic formulae) of how to determine absolute values for (17)R(VPDB CO2) and (17)R(VSMOW). Here we present the results of experiments conducted to determine these values. Two oxygen gases (one depleted in heavy isotopes and the other isotopically normal oxygen) were analysed to obtain the relative (17)O content. Samples of both gases were converted into CO(2), and the resulting CO(2) samples were analysed as well. Possible experimental and analytical errors are carefully considered and eliminated as far as feasible. Much attention was paid to understanding and dealing with cross-contamination effects occurring in the mass spectrometer. Based on the data obtained, the absolute values are calculated to be: (17)R(VPDB-CO2) = 0.00039511 +/- 0.00000094 and (17)R(VSMOW) = 0.00038672 +/- 0.00000087 (expanded uncertainties). Both values are on the original scale of Craig (Geochim. Cosmochim. Acta 1957; 12: 133-149) with (13)R(VPDB-CO2) = 0.0112372. A (17)O-correction algorithm incorporating the newly determined value for (17)R(VPDB-CO2) and lambda = 0.528 by Meijer and Li (Isot. Environ. Health Stud. 1998; 34: 349-369) is constructed. A computational test is performed to demonstrate the degree of delta(13)C bias relative to the previously known correction algorithms. delta(13)C values produced by the constructed algorithm are in the middle of the values produced by the other algorithms. We refrain, however, from giving any recommendation concerning which (17)O-correction algorithm to use in order to obtain delta(13)C data in the most accurate way. The present work illuminates the need to reconsider recommendations concerning the correction algorithm. PMID- 12720282 TI - Isomer differentiation by combining gas chromatography, selective self ion/molecule reactions and tandem mass spectrometry in an ion trap mass spectrometer. AB - This study presents a novel, simple and rapid procedure for isomer differentiation by combining gas chromatography (GC), a selective self ion/molecule reaction (SSIMR) and tandem mass spectrometry (MS/MS) in an ion trap mass spectrometer (ITMS). SSIMR product ions were produced from four isomers. For aniline, SSIMR induces the formation of the molecular ion, [M+H](+), [M+CH](+), adduct ions of fragments ([M+F](+), where F represents fragment ions) and [2M H](+). 2 and 3-Picoline produce [M+H](+), [2M-H](+) and [M+F](+), while 5 hexynenitrile produces [M+H](+), [M+F](+) and [2M+H](+) ions. The proposed method provides a relatively easy, rapid and efficient means of isomer differentiation via a SSIMR in the ITMS. Typically, isomer differentiation can be achieved within several minutes. The superiority of the SSIMR technique for isomer differentiation over electronic ionization (EI) is also demonstrated. PMID- 12720284 TI - On-column digestion of proteins in aqueous-organic solvents. AB - Proteolytic digestion is an important step in protein identification by peptide mass mapping and tandem mass spectrometry (MS/MS)-based peptide sequencing. Traditional methods of protein digestion require extended incubation times and have difficulty with proteolytically resistant proteins. Here, we describe a method in which a protein solution was combined with a mixed aqueous-organic solution (methanol, isopropanol, or acetonitrile) and passed through a microcolumn containing immobilized trypsin. Myoglobin sequence coverage was high (>85%) in all three solvents, and differences in spectra were seen among the different solution conditions. Notably, methanol-based digestions produced fewer missed cleavages while acetonitrile-based digestions produced the most peptides and the most intense mass spectra. Flow rates through the column were varied from 0.5 to 15 micro L/min, corresponding to column residence times of 78 and 2.6 s, respectively. All flow rates produced high sequence coverage of myoglobin, although, at higher flow rates, more missed cleavages were observed. No significant increase in undigested myoglobin was observed with flow rates up to 15 micro L/min. The described method was applied to the digestion of human transferrin (hTf), a proteolytically resistant protein. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometric (MALDI-TOFMS) analysis detected 42 peptides covering 46% of the hTf sequence. The traditional aqueous method resulted in 12 peptides (8% sequence coverage) only when high concentrations of trypsin were used. Lastly, digestion of low nanomolar myoglobin was shown to produce detectable peptides and resulted in a correct database hit. Thus, we demonstrate a method that is capable of rapid on-line digestion, thereby lending itself to high-throughput identification of proteins. PMID- 12720283 TI - Liquid chromatography/tandem mass spectrometry for the identification and determination of trichothecenes in maize. AB - A reliable, sensitive and selective liquid chromatography/tandem mass spectrometry (LC/MS/MS) method has been developed to determine four trichothecene mycotoxins (nivalenol, deoxynivalenol, fusarenon X and 3-acetyldeoxynivalenol) in maize. Sample preparation was performed by extracting the analytes with a mixture of acetonitrile and water, followed by a solid-phase extraction with Carbograph-4 cartridges as the purification step. For the LC/MS/MS analysis two interfacing systems, Turbo IonSpray (TISP) and atmospheric pressure chemical ionization (APCI), were compared in both negative and positive ion modes. LC and MS parameters were optimized to achieve better results and sensitivity. The effect of mobile phase modifiers such as ammonium acetate and formic acid on the ionization yield was also evaluated. The best results were obtained using the electrospray ionization (ESI) interface in negative ion mode and the multiple reaction monitoring mode (MRM) for the quantitation. The detection limits ranged between 10 ng/g for fusarenon X and 1.5 ng/g for deoxynivalenol. A linear working range was achieved with a standard deviation between 3 and 10% and recovery rates from the maize samples above 81%. The procedure was applied to the analysis of a set of maize samples collected from farms located in different areas of northern and central Italy. The investigated samples turned out to be contaminated primarily with deoxynivalenol and, to a minor extent, with its derivatives. PMID- 12720285 TI - Asymmetrical features of mass spectral peaks produced by quadrupole mass filters. AB - The individual mass spectral peaks produced by a quadrupole mass spectrometer (QMS) are asymmetric; they exhibit a 'tail' on the low mass side. In some cases a definite structure is observed in the tail. We show that the tail structure is a consequence of the use of circular electrodes. An extreme case of an experimentally observed QMS mass peak with a distinct tail structure is shown and the general form is reproduced using our numerical model. The effect of instrument resolution, length, operating frequency, ion energy, mass and ion source aperture upon the tail structure are considered. Results show that extensive long tails originate mainly from ions that enter the mass filter at a relatively large distance from the QMS axis; also no significant tail is produced in the case of ideal hyperbolic form electrodes of finite length. PMID- 12720286 TI - Product ion scanning using a Q-q-Q linear ion trap (Q TRAP) mass spectrometer. AB - The use of a Q-q-Q(linear ion trap) instrument to obtain product ion spectra is described. The instrument is based on the ion path of a triple quadrupole mass spectrometer with Q3 operable as either a conventional RF/DC quadrupole mass filter or a linear ion trap mass spectrometer with axial ion ejection. This unique ion optical arrangement allows de-coupling of precursor ion isolation and fragmentation from the ion trap itself. The result is a high sensitivity tandem mass spectrometer with triple quadrupole fragmentation patterns and no inherent low mass cut-off. The use of the entrance RF-only section of the instrument as accumulation ion trap while the linear ion trap mass spectrometer is scanning enhances duty cycles and results in increased sensitivities by as much as a factor of 20. The instrument is also capable of all of the triple quadrupole scans including multiple-reaction monitoring (MRM) as well as precursor and constant neutral loss scanning. The high product ion scanning sensitivity allows the recording of useful product ion spectra near the MRM limit of quantitation. PMID- 12720287 TI - Investigation of EDTA anticoagulant in plasma to improve the throughput of liquid chromatography/tandem mass spectrometric assays. AB - In this study, EDTA and heparin are compared as anticoagulants with respect to their efficiency in preventing clot formation in plasma samples that were subsequently analyzed by liquid chromatography/tandem mass spectrometry (LC/MS/MS). A pilot in vivo pharmacokinetic study for the drug chlorpheniramine was conducted in which both EDTA and heparin plasma samples were collected simultaneously. All conditions except the anticoagulant were held constant during the pharmacokinetic study. Bioanalytical results were compared from samples transferred by manual pipette and by an automated liquid handler workstation. The concentration of chlorpheniramine in samples was determined by LC/MS/MS. Results from the analysis of variances (ANOVA) of log-transformed plasma chlorpheniramine concentrations were used to calculate 90% confidence intervals for the ratio least-squares mean values for anticoagulants and for transfer methods. Analytical concentrations of the drug chlorpheniramine were equivalent in heparin- and EDTA containing plasma. Results suggest that the failure rate for transfer of EDTA plasma (50 micro L by automated workstation or manually) is less than that for heparinized plasma. As a consequence of these results, the vast majority of plasma samples in our laboratories are now collected in EDTA, which allows for use of automated sample transfer resulting in a three-fold timesaving over manual transfer using a single-channel pipette. The ability to use automation has resulted in improved efficiency and cost savings. PMID- 12720288 TI - Optimization of in-gel protein digestion system in combination with thin-gel separation and negative staining in 96-well plate format. AB - Improvement of in-gel digestion efficiency is highly desirable for one- or two dimensional gel electrophoretic separation and mass spectrometric (MS) analysis in proteomics, because the resultant increases in sequence coverage and MS signal intensity lead to higher confidence in protein identification. Here an optimized in-gel digestion system, in combination with thin-gel separation and negative staining in a high-throughput format using 96-well plates, is described. The combination of negative staining and protein separation on a 0.9 mm thick gel showed a clear improvement in in-gel digestion efficiency in comparison with the more typical protocols such as the combination of silver staining and a 1.0 mm gel. In addition, the use of 96-well plates to increase throughput did not decrease the efficiency of this strategy when the stirring of the gel pieces in processes such as destaining, washing, gel-shrinking and peptide extraction was performed by sonication instead of shaking the plates. This procedure was optimized and applied to identify proteins of the postsynaptic density fraction; 105 proteins were identified after SDS-PAGE separation. PMID- 12720290 TI - Reactivity of some novel multifunctional benzoyl esters towards nucleophiles investigated by electrospray ionization tandem mass spectrometry. PMID- 12720289 TI - Characterization of macrocyclic polysaccharides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. PMID- 12720292 TI - Fiber introduction mass spectrometry: coupling solid-phase microextraction with sol-gel-assisted laser desorption/ionization time-of-flight mass spectrometry. PMID- 12720291 TI - Proton affinity differences among three N-acetylhexosamines studied by the kinetic method. PMID- 12720293 TI - Evidence for involvement of the backbone in the ionization process of nucleic acids by matrix-assisted laser desorption/ionization. PMID- 12720294 TI - Overexpression of the immunoglobulin superfamily members CDO and BOC enhances differentiation of the human rhabdomyosarcoma cell line RD. AB - Rhabdomyosarcoma is a childhood tumor of the skeletal muscle lineage in which cells display defects in both biochemical and morphological aspects of differentiation. The immunoglobulin superfamily members CDO and BOC are components of a cell surface receptor that positively regulates myogenesis in vitro. Expression of Cdo and Boc in myoblast cell lines is downregulated by the ras oncogene, and forced re-expression of either Cdo or Boc can override ras induced inhibition of myogenic differentiation [Kang et al., J Cell Biol 1998; 143:403-413; Kang et al., EMBO J 2002; 21:114-124]. The current study sought to test whether the promyogenic properties of CDO and BOC could be extended to a human rhabdomyosarcoma cell line, RD. Stable overexpression of CDO or BOC in RD cells led to enhanced expression of two markers of muscle cell differentiation, troponin T and myosin heavy chain, and to increased formation of elongated, myosin heavy chain-positive myotubes. These observations are consistent with the notion that CDO and BOC play a role in the inverse relationship between differentiation and transformation of cells in the skeletal muscle lineage. PMID- 12720295 TI - Increased frequency of gastrin-releasing peptide receptor gene mutations during colon-adenocarcinoma progression. AB - Epithelial cells lining the mature human colon do not normally express receptors for gastrin-releasing peptide (GRPR). In contrast, we have shown that when aberrantly expressed in functional form in colon cancer, this protein acted as a morphogen where it caused tumor cells to adopt a better-differentiated phenotype. Importantly, GRPR mRNA is ubiquitously mutated in human colon cancer cell lines, with inactivating mutations detected in all cell lines not expressing functional receptor. Since colon cancers are heterogeneously differentiated, we set out to determine if the GRPR gene was mutated as a function of tumor cell differentiation in archived human colon cancers. We used laser capture microscopy to dissect out 67 regions of defined differentiation from 20 human colon cancers randomly selected from the UIC GI Tumor Bank. Except for two polymorphisms, the GRPR gene was not mutated in nonmalignant epithelial cells. In contrast, 42 distinct mutations were identified in malignant cells. Overall mutation number inversely correlated with the degree of tumor cell differentiation. Within any cancer, all GRPR mutations found within better-differentiated cells were conserved in more poorly-differentiated cells; while all poorly-differentiated cells contained mutations resulting in GRPR pharmacological inactivation. These data suggest that accumulation of mutations within the GRPR gene ultimately resulting in the production of nonfunctional receptors may represent a previously unappreciated mechanism allowing for the dedifferentiation of tumor cells within any particular colon cancer; and that poorly-differentiated tumor cells within any individual cancer may arise clonally from their better-differentiated precursors. PMID- 12720296 TI - Involvement of c-jun N-terminal kinase activation in 15-deoxy-delta12,14 prostaglandin J2-and prostaglandin A1-induced apoptosis in AGS gastric epithelial cells. AB - Cyclopentenone prostaglandins (CyPGs), derivatives of arachidonic acid, have been suggested to exert growth-inhibitory activity through peroxisome proliferator activated receptor (PPAR)-dependent and -independent mechanisms. Here we examined various eicosanoids for growth inhibition and found that the terminal derivative of prostaglandin (PG) J(2) metabolism, 15-deoxy-Delta(12,14)-PGJ(2) (15d-PGJ(2)), and PGA(1) markedly inhibited the growth and induced apoptosis in AGS gastric carcinoma cells. There were no significant increases in cell death and DNA fragmentation in the cells with overexpression of PPARalpha or PPARgamma, indicating the possibility that 15d-PGJ(2) and PGA(1) induced apoptosis through PPAR-independent pathway. Moreover, 15d-PGJ(2) and PGA(1) activated the c-jun N terminal kinase (JNK) and caspase-3 activity in dose- and time-dependent manners. To examine further the role of JNK signaling cascades in apoptosis induced by 15d PGJ(2) and PGA(1), we transfected dominant-negative (DN) mutants of JNK plasmid into the cells to analyze the apoptotic characteristics of cells overexpressing DN-JNK following exposure to 15d-PGJ(2) and PGA(1). Overexpression of DN-JNK significantly repressed both endogenous JNK and caspase-3 activity, and subsequently decreased apoptosis induced by 15d-PGJ(2) and PGA(1). These results suggested that CyPGs, such as 15d-PGJ(2) and PGA(1), activated JNK signaling pathway, and that JNK activation may be involved in 15d-PGJ(2)- and PGA(1) induced apoptosis. PMID- 12720297 TI - Variation in cyclooxygenase expression levels within the colorectum. AB - The positive association of decreased risk of colorectal cancer with nonsteroidal antiinflammatory drug (NSAID) use, combined with the observation that cyclooxygenase(COX)-2 is present in a majority of colorectal tumors, has led to the proposed use of isozyme-specific COX inhibitors as preventive agents in polyp and tumor formation in the colon. However, the exact biochemical mechanisms and disease stage at which reduced risk is mediated remain somewhat controversial, in part because of the complex biochemical changes that occur during the progression from aberrant crypt to polyp to tumor. In this study, COX-1 and COX-2 protein expression levels were determined in sets of tumor and normal colon tissue. Changes were characterized in COX-1 and COX-2 expression within individuals, in relation to such factors as sex, tumor grade, and location in the colorectum. COX 1 expression levels were found to be significantly reduced in tumors compared to matched normal tissues (Dunn's method, P < 0.05). Additionally, COX-1 expression was decreased in stage T3 tumors as compared to stage T2 tumors (Student's t test, P = 0.009). Similar to previous reports, COX-2 protein expression was present in 73% of the tumors studied and appeared to be independent of tumor grade and sex. Interestingly, decreased COX-2 expression correlated with tumor occurrence in rectal mucosa (Wilcoxon two-sample test, P < 0.05). These results warrant further investigation, especially the identification of determinants that would predict which populations would be most responsive to COX-2 inhibition as a means of colorectal cancer chemoprevention. PMID- 12720298 TI - Alteration of O6-methylguanine-DNA methyltransferase in colorectal neoplasms in sporadic and familial adenomatous polyposis patients. AB - DNA repair failure is known to be a critical event during carcinogenesis of colorectal cancers. To investigate whether O(6)-methylguanine-DNA methyltransferase (MGMT) is altered during colorectal carcinogenesis, we performed immunohistochemical staining on 265 sporadic colorectal cancers, 113 sporadic adenomas, 33 familial adenomatous polyposis (FAP) colorectal cancers, and 93 FAP adenomas. Sixty-seven of 265 sporadic colorectal cancer cases and five of 113 sporadic adenoma cases showed loss of MGMT expression (P < 0.001). Among FAP patients, four of 33 cancers and six of 93 adenomas showed loss of MGMT protein expression. When we compared the association between MGMT promoter hypermethylation and protein expression, almost all cases without a methylated allele were positive for the expression of MGMT. In contrast, cases with promoter methylation frequently showed loss of MGMT expression (P < 0.01). Loss of MGMT was correlated with some clinicopathological characteristics, i.e., tumor invasion (P = 0.013) and stage (P = 0.035) in sporadic colorectal cancer, and degree of atypism (P = 0.042) in sporadic adenoma. Our results show that loss of expression of MGMT occurs more frequently in cancer than in adenoma in both sporadic and FAP patients, and that loss of expression of MGMT is associated with hypermethylation of the promoter area of MGMT gene. PMID- 12720299 TI - Molecular mechanisms of G0/G1 cell-cycle arrest and apoptosis induced by terfenadine in human cancer cells. AB - Terfenadine (TF), a highly potent histamine H1 receptor antagonist, has been shown to exert no significant central nervous system side effects in clinically effective doses. In this study, we demonstrated that TF induced significant growth inhibition of human cancer cells, including Hep G2, HT 29, and COLO 205 cells, through induction of G(0)/G(1) phase cell-cycle arrest. The minimal dose of TF induced significant G(0)/G(1) arrest in these cells was 1-3 microM. The protein levels of p53, p21/Cip1, and p27/Kip1 were significantly elevated, whereas the kinase activities of cyclin-dependent kinase 2 (CDK2) and CDK4 were inhibited simultaneously in the TF-treated cells. On the other hand, significant apoptosis, but not G(0)/G(1) arrest, was induced in the HL 60 (p53-null) or Hep 3B (with deleted p53) cells when treated with TF (3-5 microM). To clarify the roles of p21/Cip1 and p27/Kip1 protein expression, which was involved in G(0)/G(1) arrest and apoptosis induced by TF in human cancer cells, antisense oligodeoxynucleotides (ODNs) specific to p21/Cip1 and p27/Kip1 were used, and the expression of the p21/Cip1 and p27/Kip1 were monitored by immunoblotting analysis. Our data demonstrated that the percentage of the apoptotic cells detected by annexin V/PI analysis in the TF-treated group was clearly attenuated by pretreatment with p27/Kip1-specific ODNs. These results indicated that p27/Kip1 (but not p21/Cip1) protein indeed played a critical role in the TF induced apoptosis. We also demonstrated that the TF-induced G(0)/G(1) cell-cycle arrest effect was not reversed by TF removal, and this growth inhibition lasted for at least 7 d. Importantly, the occurrence of apoptosis and cell growth arrest was not observed in the TF-treated normal human fibroblast, even at a dose as high as 25 microM. Our study showed the molecular mechanisms for TF-induced cell growth inhibition and the occurrence of apoptosis in human cancer cells. PMID- 12720300 TI - Defective processing of the transforming growth factor-beta1 in azoxymethane induced mouse colon tumors. AB - High levels of the cell growth inhibitor transforming growth factor-beta1 (TGF beta1) are often found in a variety of human cancers. However, the physiological significance of this overexpression depends on the availability of the biologically active form of TGF-beta1 within the extracellular matrix of the tumor microenvironment. To determine the expression and activation status of TGF beta1 in chemically induced tumors, 6-wk-old A/J mice were injected intraperitoneally with either azoxymethane (AOM) (10 mg/kg body weight, once a week for 6 wk) or normal saline solution, and colon tumors were isolated 24 wk following the last injection. An enzyme-linked immunosorbent assay for TGF-beta1 revealed a significant increase (1.7-fold, P < 0.05) in total TGF-beta1 protein in tumors. Interestingly, while 80% of the total TGF-beta1 in the control colon tissues was in the active form, only 50% was found to be active in tumors. Together with our earlier observations that TGF-beta1 mRNA levels are unchanged in A/J tumors, these data further support a mechanism whereby elevated TGF-beta1 levels result from a defective activation and turnover of this protein. Because plasmin is known to be a major activator of TGF-beta1 in vivo, we hypothesized that reduced plasmin activity may be responsible for the observed dysregulation of TGF-beta1 processing in these behaviorally benign tumors. With a fluorogenic peptide substrate for serine proteases, a deficiency in plasmin activity was found in the tumors. Furthermore, semiquantitative reverse transcription (RT) polymerase chain reaction (PCR) analysis of a panel of genes involved in the plasminogen activation system, including plasminogen activator inhibitor-1 (PAI 1), urokinase-plasminogen activator (u-PA), and urokinase-receptor (u-PAR-1), demonstrated a significant upregulation (approximately fourfold to sixfold, P < 0.05) in the expression of each of these genes in the tumor tissue. In addition, no significant changes were observed in the expression levels of thrombospondin-1 (TSP-1) and insulin-like growth factor type II receptor (IGF-IIR), which also mediate the activation of latent TGF-beta1. To gain further insight into the functionality of the TGF-beta1 pathway, cDNA microarrays were performed and the expression levels of a panel of 21 TGF-beta1-specific target genes were determined in AOM-induced tumors that overexpress the ligand. A significant dysregulation in the expression of each of these targets was observed, providing evidence of aberrant TGF-beta1 signaling in tumors. Overall, the present study demonstrates a very low plasmin activity in A/J colon tumors, possibly as a result of the potent inhibitory effect of PAI-1 on the plasminogen activation cascade. The observed deficiency in plasmin activity may not be sufficiently compensated for by other mechanisms of latent TGF-beta1 activation, including TSP 1 and IGF-IIR, thereby resulting in a decreased fraction of the biologically active form of TGF-beta1 and subsequent aberration in TGF-beta1-specific gene regulation in A/J tumors. PMID- 12720301 TI - Modelling the random effects covariance matrix in longitudinal data. AB - A common class of models for longitudinal data are random effects (mixed) models. In these models, the random effects covariance matrix is typically assumed constant across subject. However, in many situations this matrix may differ by measured covariates. In this paper, we propose an approach to model the random effects covariance matrix by using a special Cholesky decomposition of the matrix. In particular, we will allow the parameters that result from this decomposition to depend on subject-specific covariates and also explore ways to parsimoniously model these parameters. An advantage of this parameterization is that there is no concern about the positive definiteness of the resulting estimator of the covariance matrix. In addition, the parameters resulting from this decomposition have a sensible interpretation. We propose fully Bayesian modelling for which a simple Gibbs sampler can be implemented to sample from the posterior distribution of the parameters. We illustrate these models on data from depression studies and examine the impact of heterogeneity in the covariance matrix on estimation of both fixed and random effects. PMID- 12720302 TI - Approximate cross-validatory predictive checks in disease mapping models. AB - When fitting complex hierarchical disease mapping models, it can be important to identify regions that diverge from the assumed model. Since full leave-one-out cross-validatory assessment is extremely time-consuming when using Markov chain Monte Carlo (MCMC) estimation methods, Stern and Cressie consider an importance sampling approximation. We show that this can be improved upon through replication of both random effects and data. Our approach is simple to apply, entirely generic, and may aid the criticism of any Bayesian hierarchical model. PMID- 12720303 TI - Bayesian evaluation of breast cancer screening using data from two studies. AB - The mean sojourn time (the duration of the period during which a cancer is symptom free but potentially detectable by screening) and the screening sensitivity (the probability that a screen applied to a cancer in the preclinical screen detectable period will result in a positive diagnosis) are two important features of a cancer screening programme. Little data from any single study are available on the potential effectiveness of mammographic screening for breast cancer in women with a family history of the disease, despite this being an important public health issue. We develop a method of estimation, from two separate studies, of the two parameters, assuming that transition from no disease to preclinical screen detectable disease, and from preclinical disease to clinical disease, are Poisson processes. Estimation is performed by a Markov chain Monte Carlo algorithm. The method is applied to the synthesis of two studies of mammographic screening in women with a family history of breast cancer, one in Manchester and one in Kopparberg, Sweden. PMID- 12720304 TI - Markov modelling of changes in HIV-specific cytotoxic T-lymphocyte responses with time in untreated HIV-1 infected patients. AB - HIV-specific cytotoxic CD8(+) T-lymphocytes (CTL) appear to be the cornerstone of the immune response to HIV infection. Recent studies show that CTL activity reflects patients' anti-HIV immune status and slows disease progression. However, the dynamics of the diversity of this response also appears as a key parameter for immune control but the dynamics of this diversity is largely undocumented. We modelled changes in CTL responses against the seven principal HIV proteins over time. We also studied the influence of plasma viral load on temporal changes in HIV protein recognition by memory CTL. The generic model we developed is based on a continuous time homogeneous Markov process with reversible states. Those states are defined by the number of proteins recognized by memory CTL in a given patient at a given time. This approach was developed within a Bayesian framework. Full Bayesian inference is implemented using Markov chain Monte Carlo simulations (MCMC). The Gibbs sampling algorithm was used to estimate the marginal posterior distributions of the transition intensities between stages of CTL responses. We applied our model to data of 152 HIV-infected patients included in the IMMUNOCO cohort. The model suggested that the diversity of HIV protein recognition by memory CTL in treatment-naive patients decreases as the disease progresses. Namely, the loss of T cytotoxic responses is globally faster than their acquisition. Indeed, these patients' T cytotoxic responses were characterized by marked individual turnover and a gradual loss of multiple protein recognition over time, this loss accelerating as viral load increased. PMID- 12720305 TI - Exploring heterogeneity in tumour data using Markov chain Monte Carlo. AB - We describe a Bayesian approach to incorporate between-individual heterogeneity associated with parameters of complicated biological models. We emphasize the use of the Markov chain Monte Carlo (MCMC) method in this context and demonstrate the implementation and use of MCMC by analysis of simulated overdispersed Poisson counts and by analysis of an experimental data set on preneoplastic liver lesions (their number and sizes) in the presence of heterogeneity. These examples show that MCMC-based estimates, derived from the posterior distribution with uniform priors, may agree well with maximum likelihood estimates (if available). However, with heterogeneous parameters, maximum likelihood estimates can be difficult to obtain, involving many integrations. In this case, the MCMC method offers substantial computational advantages. PMID- 12720306 TI - Estimation of infection and recovery rates for highly polymorphic parasites when detectability is imperfect, using hidden Markov models. AB - A Bayesian hierarchical model is proposed for estimating parasitic infection dynamics for highly polymorphic parasites when detectability of the parasite using standard tests is imperfect. The parasite dynamics are modelled as a non homogeneous hidden two-state Markov process, where the observed process is the detection or failure to detect a parasitic genotype. This is assumed to be conditionally independent given the hidden process, that is, the underlying true presence of the parasite, which evolves according to a first-order Markov chain. The model allows the transition probabilities of the hidden states as well as the detectability parameter of the test to depend on a number of covariates. Full Bayesian inference is implemented using Markov chain Monte Carlo simulation. The model is applied to a panel data set of malaria genotype data from a randomized controlled trial of bed nets in Tanzanian children aged 6-30 months, with the age of the host and bed net use as covariates. This analysis confirmed that the duration of infections with parasites belonging to the MSP-2 FC27 allelic family increased with age. PMID- 12720307 TI - A mixture model for bovine abortion and foetal survival. AB - The effect of spontaneous abortion on the dairy industry is substantial, costing the industry on the order of US dollars 200 million per year in California alone. We analyse data from a cohort study of nine dairy herds in Central California. A key feature of the analysis is the observation that only a relatively small proportion of cows will abort (around 10;15 per cent), so that it is inappropriate to analyse the time-to-abortion (TTA) data as if it were standard censored survival data, with cows that fail to abort by the end of the study treated as censored observations. We thus broaden the scope to consider the analysis of foetal lifetime distribution (FLD) data for the cows, with the dual goals of characterizing the effects of various risk factors on (i). the likelihood of abortion and, conditional on abortion status, on (ii). the risk of early versus late abortion. A single model is developed to accomplish both goals with two sets of specific herd effects modelled as random effects. Because multimodal foetal hazard functions are expected for the TTA data, both a parametric mixture model and a non-parametric model are developed. Furthermore, the two sets of analyses are linked because of anticipated dependence between the random herd effects. All modelling and inferences are accomplished using modern Bayesian methods. PMID- 12720308 TI - A meta-analysis of studies on the association of the platelet PlA polymorphism of glycoprotein IIIa and risk of coronary heart disease. AB - The Pl(A2) polymorphism of the glycoprotein IIIa subunit of the fibrinogen receptor (GPIIb-IIIa) has been reported by some studies to be associated with an increased risk of coronary thrombosis. Following the first paper on the subject in 1996, a large number of studies have investigated the relationship between this polymorphism and coronary thrombosis, either at the epidemiological or at the cellular and molecular levels. The cellular and molecular studies have shown in a consistent manner that this polymorphism increases platelet responsiveness. In contrast, epidemiological studies have generated inconsistent results regarding the clinical impact of Pl(A2). We consider 12 epidemiological studies that investigate the link between presence/absence of this polymorphism and presence/absence of coronary heart disease. Each is a case-control study that reports an odds ratio. The studies are not directly comparable because they differ greatly in their patient pools and also in the way the data are analysed. We present several meta-analyses of these 12 studies. The simplest one is based on a standard frequentist random effects model with a normal distribution for the study effects (the per-study population log-odds ratios). We also consider a Bayesian version of this model, with a diffuse prior for the mean and variance of the normal distribution of the study effects. The conclusions from both of these analyses is about the same, and is that there is evidence that the Pl(A2) polymorphism is associated with an increased risk of coronary heart disease. A look at the reported log-odds ratios across studies suggests that the study effects do not come from a symmetric distribution. For this reason, we also consider semi-parametric priors for the distribution of the study effects. These priors are specifically designed for this kind of meta-analysis, and are based on a certain class of mixtures of Dirichlet priors. They can be designed to concentrate most of their mass around the family of normal distributions, but still allow for any other distribution. The semi-parametric Bayesian model continues to give evidence of an association between the Pl(A2) polymorphism and the risk of coronary heart disease. PMID- 12720309 TI - Hierarchical Bayesian spatial modelling of small-area rates of non-rare disease. AB - We present Bayesian hierarchical spatial models for the analysis of the geographical distribution of a non-rare disease or event. The work is motivated by the need for ascertaining regional variations in health services outcomes and resource use and for assessing the potential sources of these variations. The models discussed herein readily accommodate random spatial effects and covariate effects. We discuss Bayesian inferential framework and implementation of a hybrid Markov chain Monte Carlo method for full Bayesian model inference. The methods are illustrated through an analysis of regional variation in chronic lung disease (CLD) rates among neonatal intensive care unit (NICU) patients across Canada. Specifically, we first present a random effects binomial model for spatially correlated CLD rates, with random spatial effects accounting for latent or covariate effects. These random spatial effects depict regional or spatial variation in chronic lung disease occurrence. We then extend this model to include covariates. With this extension, we assess residual spatial effects and the extent to which risk factors such as illness severity at NICU admission, low birth weight, and very low birth weight influence the CLD rate variation. PMID- 12720311 TI - A computational study of conformational interconversions in 1,4-dithiacyclohexane (1,4-dithiane). AB - Ab initio molecular orbital theory with the 6-31G(d), 6-31G(d,p), 6-31+G(d), 6 31+G(d,p), 6-31+G(2d,p), 6-311G(d), 6-311G(d,p), and 6-311+G(2d,p) basis sets and density functional theory (BLYP, B3LYP, B3P86, B3PW91) have been used to locate transition states involved in the conformational interconversions of 1,4 dithiacyclohexane (1,4-dithiane) and to calculate the geometry optimized structures, relative energies, enthalpies, entropies, and free energies of the chair and twist conformers. In the chair and 1,4-twist conformers the C-Hax and C Heq bond lengths are equal at each carbon, which suggest an absence of stereoelectronic hyperconjugative interactions involving carbon-hydrogen bonds. The 1,4-boat transition state structure was 9.53 to 10.5 kcal/mol higher in energy than the chair conformer and 4.75 to 5.82 kcal/mol higher in energy than the 1,4-twist conformer. Intrinsic reaction coordinate (IRC) calculations showed that the 1,4-boat transition state structure was the energy maximum in the interconversion of the enantiomers of the 1,4-twist conformer. The energy difference between the chair conformer and the 1,4-twist conformer was 4.85 kcal/mol and the chair-1,4-twist free energy difference (deltaG degrees (c-t)) was 4.93 kcal/mol at 298.15 K. Intrinsic reaction coordinate (IRC) calculations connected the transition state between the chair conformer and the 1,4-twist conformer. This transition state is 11.7 kcal/mol higher in energy than the chair conformer. The effects of basis sets on the 1,4-dithiane calculations and the relative energies of saturated and unsaturated six-membered dithianes and dioxanes are also discussed. PMID- 12720310 TI - Comparing the importance of disease rate versus practice style variations in explaining differences in small area hospitalization rates for two respiratory conditions. AB - Many studies have reported large variations in age- and sex-adjusted rates of hospitalizations across small geographic areas. These variations have often been attributed to differences in medical practice style which are not reflected in differences in health care outcomes. There is, however, another potentially important source of variation that has not been examined much in the literature: geographic differences in the age-sex adjusted size of the pool of patients who present with the disease and are candidates for hospitalization. Previous studies of small area variations in hospitalization rates have only used data on hospitalizations. Thus, it has not been possible to distinguish the extent to which differences in hospitalization rates are due to (i). differences in the chance that patients diagnosed with a disease are admitted to a hospital, which we refer to as the 'practice style effect,' versus (ii). geographic differences in the total amount of diagnosed disease, which we refer to as the 'disease effect.' Elementary methods for estimating the relative strength of the two effects directly from the data can be misleading, since equal amounts of variability in each effect result in unequal impacts on hospitalization rates. In this paper we describe a model-based approach for estimating the relative importance of the practice style effect and the disease effect in explaining variations in hospitalization rates. The key to our approach is the use of data on both inpatient and outpatient visits. We use 1997 Medicare data for two respiratory medical conditions across 71 small areas in Massachusetts: chronic bronchitis and emphysema, and bacterial pneumonia. Based on a Poisson model for the process generating hospitalizations and outpatient visits, we use a Bayesian framework and Gibbs sampling to compute and compare the correlation between the number of people hospitalized and each of these two sources of variation. Our results show that for the two conditions, disease rate variation explains at least as much of the variation in hospitalization rates as does practice style variation. PMID- 12720312 TI - Rigid-body dynamics in the isothermal-isobaric ensemble: a test on the accuracy and computational efficiency. AB - We have developed a time-reversible rigid-body (rRB) molecular dynamics algorithm in the isothermal-isobaric (NPT) ensemble. The algorithm is an extension of rigid body dynamics [Matubayasi and Nakahara, J Chem Phys 1999, 110, 3291] to the NPT ensemble on the basis of non-Hamiltonian statistical mechanics [Martyna, G. J. et al., J Chem Phys 1994, 101, 4177]. A series of MD simulations of water as well as fully hydrated lipid bilayer systems have been undertaken to investigate the accuracy and efficiency of the algorithm. The rRB algorithm was shown to be superior to the state-of-the-art constraint-dynamics algorithm SHAKE/RATTLE/ROLL, with respect to computational efficiency. However, it was revealed that both algorithms produced accurate trajectories of molecules in the NPT as well as NVT ensembles, as long as a reasonably short time step was used. A couple of multiple time-step (MTS) integration schemes were also examined. The advantage of the rRB algorithm for computational efficiency increased when the MD simulation was carried out using MTS on parallel processing computer systems; total computer time for MTS-MD of a lipid bilayer using 64 processors was reduced by about 40% using rRB instead of SHAKE/RATTLE/ROLL. PMID- 12720313 TI - Ab initio studies for the photodissociation mechanism of hydroxyacetone. AB - The reaction pathways for CH(3)COCH(2)OH (hydroxyacetone) photodissociation on the low-lying electronic states have been studied with use of the CASSCF energy gradient techniques. The S(0)/S(1) and S(1)/T(1) intersection points were determined by the state-average CASSCF method. Two main reaction pathways, which are possible to the photodissociation, have been studied. It has been found that the mechanism is stepwise, and belongs to Norrish type-I reaction. The n --> pi* excitation leads to the first excited singlet state, followed by the intersystem crossing from S(1) to T(1). On the T(1) potential energy surface, the system can dissociate adiabatically to CH(3)(x) +COCH(2)OH( x) and CH(3)CO(x)+CH(2)OH(x). The COCH(2)OH(x) and CH(3)CO(x) radicals can further dissociate into CO, OH, and other fragments. Our calculated results are in good agreement with recent experimental results. PMID- 12720314 TI - A computational study of interactions between acetic acid and water molecules. AB - Density functional theory calculations were performed for the title reactions to elucidate the difference between the strong cyclic hydrogen bond of (Me-COOH)(2) and the electrolytic dissociation, MeCOOH <==> Me-COO(-) + H(+), as a weak acid. The association of water clusters with acetic acid dimers strengthens the cyclic hydrogen bond. A nucleophilic attack of the carboxylic carbon by a water cluster leads to a first zwitterionic intermediate, MeCOO(-) + H(3)O(+) + (HO)(3)C-Me. The intermediate is unstable and is isomerized to a neutral interacting system, MeCOOH...(HO)(3)C-Me + H(2)O. The ethanetriol, (HO)(3)-CMe is transformed to an acetic acid monomer. The monomer may be dissociated to give a second zwitterionic intermediate with reasonable proton-relay patterns and energy changes. In proton relay reaction channels, H in MeCOOH is not an acidic proton but is always a hydroxy proton. PMID- 12720315 TI - The structure and stability of Si60 and Ge60 cages: a computational study. AB - Structural studies of fullerene-like Si(60) and Ge(60) cages using ab initio methods were augmented by density functional tight-binding molecular dynamics (DFTB-MD) simulations of finite temperature effects. Neither the perfect I(h) symmetry nor the distorted T(h) structures are true minima. The energies of both are high relative to distorted, lower symmetry minima, C(i) and T, respectively, which still preserve C(60)-type connectivity. Both Si(60) and Ge(60) favor C(i) symmetry cages in which Si and Ge vertexes exhibit either near-trigonal or pyramidal geometries. These structural variations imply significant reactivity differences between different positions. The small magnetic shielding effects (NICS) indicate that aromaticity is not important in these systems. The inorganic fullerene cages have lower stabilities compared with their carbon analogs. Si(60) is stable towards spontaneous disintegration up to 700 K according to DFTB-MD simulations, and thus has potential for experimental observation. In contrast, Ge(60) preserves its cage structure only up to 200 K. PMID- 12720316 TI - On the transferability of hydration-parametrized continuum electrostatics models to solvated binding calculations. AB - Using molecular mechanics force field partial atomic charges, we show the nonuniqueness of the parametrization of continuum electrostatics models with respect to solute atomic radii and interior dielectric constant based on hydration (vacuum-to-water transfer) free energy data available for small molecules. Moreover, parameter sets that are optimal and equivalent for hydration free energy calculations lead to large variations of calculated absolute and relative electrostatic binding free energies. Hence, parametrization of solvation effects based on hydration data, although a necessary condition, is not sufficient to guarantee its transferability to the calculation of binding free energies in solution. PMID- 12720317 TI - Theoretical studies on the gas-phase pyrolysis of 2-phenoxycarboxylic acids: an ONIOM approach. AB - Various ONIOM combinations-ONIOM(HF/6-31G*: PM3), ONIOM(B3LYP/6-31G*: PM3), ONIOM(MP2/6-31G*: PM3), and ONIOM(MP2/6-31G*: HF/3-21G)--were applied to investigate thermal decomposition mechanisms of four 2-phenoxycarboxylic acids (2 phenoxyacetic acid, 2-phenoxypropionic acid, 2-phenoxybutyric acid, and 2 phenoxyisobutyric acid) in the gas phase. All the transition states and intermediates of the reaction paths were optimized. The reaction pathway of four reactants yielding the phenol, CO, and the corresponding carbonyl compound was characterized on the potential energy surface and found to proceed stepwise. The first step corresponds to the elimination of phenol and the formation of alpha lactone intermediate through a five-membered ring transition state, and the second step is the cycloreversion process of alpha-lactone intermediate to form CO and the corresponding carbonyl compound. The reaction pathway of latter three compounds to produce the carboxylic acid and phenol via a four-membered cyclic transition structure was also examined theoretically. Comparison with experiment indicates that the activation parameters for the fist reaction channel are accurately predicted at the ONIOM(MP2/6-31G*: HF/3-21G) level of theory. PMID- 12720318 TI - Chemometric study of liquid water simulations. I. The parameters of the TIP4P model potential. AB - The multivariate chemometric techniques two level factorial design (TLFD) and principal component analysis (PCA) were used to investigate the TIP4P model potential behavior with respect to perturbations on all intermolecular interaction parameters. The effects of these perturbations were calculated for the enthalpy of vaporization, the density, the first maximum of the radial distribution functions of the O-H and O-O pairs, and the second maximum of the radial distribution function of the O-H pair obtained from Monte Carlo simulations of liquid water at 25 degrees C. The principal effects were quantified and rationalized in terms of the pair-wise interaction potential of the TIP4P model. They also corroborate previously published sensitivity analysis results using molecular dynamics and other model potentials. In addition, significant interaction effects between some parameters of the TIP4P model potential were observed and quantified, which hardly could be obtained without such a statistic approach. These interaction effects are very regular and systematic, and their behavior has not been encountered in other chemometric studies and cannot be rationalized in terms of the functional form of the pair wise potential. PMID- 12720319 TI - About the calculation of exchange coupling constants in polynuclear transition metal complexes. AB - The application of theoretical methods based on the density functional theory with hybrid functionals provides good estimates of the exchange coupling constants for polynuclear transition metal complexes. The accuracy is similar to that previously obtained for dinuclear compounds. We present test calculations on simple model systems based on H. He and CH(2). He units to compare with Hartree Fock and multiconfigurational results. Calculations for complete, nonmodeled polynuclear transition metal complexes yield coupling constants in very good agreement with available experimental data. PMID- 12720320 TI - A flexible nudged elastic band program for optimization of minimum energy pathways using ab initio electronic structure methods. AB - A driver program for carrying out nudged elastic band optimizations of minimum energy reaction pathways is described. This approach allows for the determination of minimum energy pathways using only energies and gradient information. The driver code has been interfaced with the GAUSSIAN 98 program. Applications to two isomerization reactions and to a cluster model for H(2) desorption from the Si(100)-2 x 1 surface are presented. PMID- 12720321 TI - OPEP: a tool for the optimal partitioning of electric properties. AB - OPEP is a suite of FORTRAN programs targeted at the optimal partitioning of molecular electric properties. It includes an interactive module for the construction of Cartesian grids of points, on which either the molecular electrostatic potential or the induction energy is mapped. The generation of distributed multipoles and polarizabilities is achieved using either the formalism of the normal equations of the least-squares problem, which restates the fitting procedure in terms of simple matrix operations, or a statistical approach, which provides a pictorial description of the distributed models of multipoles and polarizabilities, thereby allowing the pinpointing of pathological cases. Molecular symmetry is accounted for by means of local atomic frames, which are generated in an automated fashion. A Tcl/Tk graphical user interface wraps the suite of programs, thereby making OPEP a user-friendly package for building models of distributed multipoles and polarizabilities. OPEP is an open-source suite of programs distributed free of charge under the GNU general public license (GPL) at http://www.lctn.uhp-nancy.fr/Opep. PMID- 12720322 TI - The evolution of the concept of medicalization in the late twentieth century. AB - This article attempts to outline a history of the critique of medicalization that developed in the 1960s in the work of Thomas Szasz, R. D. Laing, Michael Foucault, and others that was applied in their work to medical and psychiatric theory and practice, the penal system, and public health systems. This article follows the development of Foucault's own work on "governmentality" that emphasized the individualization and internalization of the themes of medicalization and the application of these ideas by his disciples to contemporary medical developments. Finally, the author explores recent historiography in the history of medicine and public health that supports this thesis of individualization that both undermines and reconfigures the older notion of medicalization. PMID- 12720323 TI - Where did the ventricular localization of mental faculties come from? AB - During the Middle Ages it was widely believed that the various mental faculties sensation, cognition, memory, and so forth-were each located in a specific part of the three ventricles that were thought to be housed in the brain. The origin of this scheme was commonly attributed to the ancient scholars, but it was rare for a specific individual to be identified as the originator. Modern researchers sometimes attribute it to Galen, but Galen was clear in his contention that the mental faculties were located in the substance of the brain rather than in the ventricles. It was only later scholars who, using Galen's anatomy as their basis, placed the mental faculties in the ventricles themselves. The system came together piece by piece over a period of centuries, probably not appearing fully in the form known to the Medieval world until the Middle Ages themselves. This article traces the development of the theory of ventricular localization of the mental faculties from the Ancient world to the early part of the Middle Ages. PMID- 12720324 TI - "What's on the worker's mind": class passing and the study of the industrial workplace in the 1920s. AB - Posing, living, and laboring as American workers, several 1920s reformist labor investigators sought to develop an alternative to Frederick Taylor's famous characterization of a typical manual laborer as mentally akin to an ox. Through their experiences as workers, they believed that they gained real if limited access to working-class psychology. Accordingly, they presented views of the worker's mind that significantly loosened the strictures of hereditarian and, especially in the case of foreign-born workers, scientific racist thought. But their efforts were shaped by their own backgrounds and biases, and by those of the academic authorities upon whose work they built, and were also mediated through the complexities of their efforts to pass across the class line. Their work finally lent itself less to the purposes of industrial democracy and reform than to those of the rising 1920s schools of industrial psychology, industrial sociology, and personnel relations. PMID- 12720325 TI - Munsterberg's nightmare: psychology and history in fin-de-siecle Germany and America. AB - This article demonstrates that Hugo Munsterberg's presidential address "Psychology and History," delivered to the American Psychological Association in 1898, should be understood in the German context of the 1890s. It constituted a response to a central feature of fin-de-siecle culture in Europe, the revolt against positivism. To be more precise, Munsterberg reacted against a new intellectual trend that was arising in Germany in the middle 1890s: the call for a historically oriented social psychology put forward by Wilhelm Dilthey-who was explicitly attacking Munsterberg's physiological conception of psychology-and new cultural historians like Karl Lamprecht and others who seemed to be putting Dilthey's program into practice. PMID- 12720328 TI - Cleavage N-terminal to proline: analysis of a database of peptide tandem mass spectra. AB - Fragmentation at the Xxx-Pro bond was analyzed for a group of peptide mass spectra that were acquired in a Finnigan ion trap mass spectrometer and were generated from proteins digested by enzymes and identified by the Sequest algorithm. Cleavage with formation of a + b + y ions occurred more readily at the Xxx-Pro bond than at other locations in these peptides, and the importance of this cleavage varied by the identity of Xxx. The most abundant Xxx-Pro relative bond cleavage ratios were observed when Xxx was Val, His, Asp, Ile, and Leu, whereas the least abundant cleavage ratios occurred when Xxx was Gly or Pro. Rationalization for these cleavage ratios at Xxx-Pro may include contribution of the Asp or His side chain to enhanced cleavage or the conformation of Pro, Gly, and the aliphatic residues Val, Ile, and Leu at the Xxx location in the Xxx-Pro bond. Although unusual fragmentation behavior has been noted for Pro-containing peptides, this analysis suggests that fragmentation at the Xxx-Pro bond is predictable and that this information may be used to improve the identification of proteins if it is incorporated into peptide sequencing algorithms. PMID- 12720329 TI - A 10-fold improvement in the precision of boron isotopic analysis by negative thermal ionization mass spectrometry. AB - Boron isotopes are potentially very important to cosmochemistry, geochemistry, and paleoceanography. However, the application has been hampered by the large sample required for positive thermal ionization mass spectrometry (PTIMS), and high mass fractionation for negative-TIMS (NTIMS). Running as BO(2)(-), NTIMS is very sensitive and requires only nanogram sized samples, but it has rather poor precision (approximately 0.7-2.0 per thousand) as a result of the larger mass fractionation associated with the relatively light ion. In contrast, running as the much heavier molecule of Cs(2)BO(2)(+), PTIMS usually achieves better precision around 0.1-0.4 per thousand. Moreover, there is a consistent 10 per thousand offset in the (11)B/(10)B ratio for NIST SRM 951 standard boric acid between the NTIMS and the certified value, but the cause of this offset is unclear. In this paper, we have adapted a technique we developed earlier to measure the (138)La/(139)La using LaO(+) (1) to improve the NTIMS technique for BO(2). We were able to correct for instrumental fractionation by measuring BO(2)( ) species not only at masses of 42 and 43, but also at 45, which enabled us to normalize (45)BO(2)/(43)BO(2) to an empirical (18)O/(16)O value. We found that both I(45)/I(42) = ((11)B(16)O(18)O/(10)B(16)O(16)O) and (I(43)/I(42))(C) = ((11)B(16)O(16)O/(10)B(16)O(16)O) vary linearly with (I(45)/I(43))(C) x 0.5 = ((11)B(16)O(18)O/(11)B(16)O(16)O) x 0.5 = (18)O/(16)O. In addition, different activators and different chemical forms of B yield different slopes for the fractionation lines. After normalizing (11)B(16)O(18)O/(11)B(16)O(16)O x 0.5 to a fixed (18)O/(16)O value, we obtained a mean (11)B/(10)B value of NIST SRM 951 that matches the NIST certified value at 4.0430 +/- 0.0015 (+/-0.36 per thousand, n = 11). As a result, our technique can achieve precision and accuracy comparable to that of PTIMS with only 1 per thousand of the sample required. This new NTIMS technique for B isotopes is critical to the studies of early solids in the solar system and individual foraminifera in sediments that require both high sensitivity and precision. PMID- 12720330 TI - Accurate mass measurement by electrospray ionization quadrupole mass spectrometry: detection of variants differing by <6 Da from normal in human hemoglobin heterozygotes. AB - Mass spectrometry has a basic limitation when human hemoglobin variants are analyzed, because it cannot resolve two globin chains that differ in mass by <6 Da. Several common beta-chain variants differ by 1 Da from normal and, hence, when present in heterozygotes, are not resolved from the normal beta-chain. Normal and variant chains appear together in the spectrum as a single entity, whose mass is the abundance weighted mean of the two chains. Here we show that such heterozygotes can be detected in 500-fold diluted blood by accurately measuring the mass of the beta-chain using an electrospray ionization quadrupole instrument and the alpha-chain for internal mass calibration. A statistical analysis of the normal beta-chain mass (n = 86) showed that the standard deviation (SD) of the mean was <+/-0.05 Da (<+/-3.2 ppm). Hence, at the 95% confidence level (+/-2 SD), an abnormal alpha- or beta-chain differing by 1 Da from normal should be detectable in a heterozygote provided its abundance is >10% of total alpha- or beta-chains, respectively. Variants whose masses lay between 1 and 4 Da from normal were detected in 19 heterozygotes. Moreover, the proportion of each variant estimated from the mass change correlated with the proportion determined by cation-exchange HPLC. Variants were assigned to the alpha- or beta chain by combining the sign of the mass change with the polarity change inferred from electrophoretic data. This procedure could be used for screening clinically significant hemoglobin variants. PMID- 12720331 TI - Time-gated pulsed glow discharge: real-time chemical speciation at the elemental, structural, and molecular level for gas chromatography time-of-flight mass spectrometry. AB - A millisecond pulsed glow discharge is used as a versatile ion source for time gated generation of elemental, structural, and molecular ions. The utility of this ion source for comprehensive chemical analysis of a series of aromatic and halogenated hydrocarbons is illustrated in this manuscript. To highlight the analytical utility of this transient ion source, it was connected to a gas chromatograph for the mass spectrometric determination of mixtures containing benzene, toluene, o-xylene, cymene, tert-butylbenzene, carbon tetrachloride, chloroform, chlorobenzene, tetrachlorethane, and dichlorobenzene. Explicit chemical analysis was accomplished by introducing the GC eluent into a pulsed glow discharge operating at a rate of 100 Hz with a 50% duty cycle. Using three independent digitizers for time-gated acquisition in three separate time regimes, nearly concurrent collection of elemental, structural, and molecular information was accomplished. In general, elemental information was obtained during the first 0.015 ms after the plasma onset; structural information, as ascertained from molecular fragmentation, was obtained during the plateau time regime when the plasma pulse is at a steady state, whereas molecular M(+) and MH(+) ions were obtained during the afterpeak time regime, that is, after the cessation of the plasma power pulse. PMID- 12720332 TI - Functional microfabricated sample targets for matrix-assisted laser desorption/ionization mass spectrometry analysis of ribonucleic acids. AB - Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is a powerful analytical tool for the structural characterization of proteins and nucleic acids. However, many proteomics or genomics methodologies that employ MALDI-MS require external sample manipulation, which limits the overall throughput of analysis. We have focused on fabricating functional MALDI sample plates that would permit the on-probe characterization of nucleic acids. Here, we present results arising from the fabrication of functional sample plates composed of poly(methyl methacrylate) (PMMA). The PMMA sample plates were fabricated by a CNC milling technique. The key structural feature of our microfabricated samples plates is the presence of individual cylindrical posts (360 microm x 360 microm), which serve as individual sample targets within the overall PMMA-based MALDI sample plate. Functionality is added to these microposts via the covalent attachment of enzymes. As an example of the applicability of these microfabricated sample plates, enzymatic digestion of ribonucleic acids was performed on probe (i.e., on the micropost) with subsequent analysis by MALDI-MS. Advantages to such an approach include a reduction in sample handling (and concomitant sample losses) and a reduction in the amount of sample required for analysis due to the small surface area of the microposts. PMID- 12720333 TI - Time-weighted average passive sampling with a solid-phase microextraction device. AB - A modified Solid-Phase Microextraction (SPME) device has been used as a passive sampler to determine the time-weighted average (TWA) concentration of volatile organic compounds (VOCs) in air. Unlike conventional sampling with SPME, in which the fiber is extended outside its needle housing, during TWA passive sampling, the fiber is retracted a known distance into its needle housing. The SPME passive sampler collects the VOCs by the mechanism of molecular diffusion and sorption on to a coated fiber as collection medium. This process has been shown to be described by Fick's first law of diffusion, whereby determination of the amounts of analytes accumulated over time enable measurement of the TWA concentration to which the sampler was exposed. A series of fibers, 100-microm poly(dimethylsiloxane), 65-microm poly(dimethylsiloxane)/divinylbenzene, and 75 microm Carboxen/poly(dimethylsiloxane), were tested for their "zero sink", face velocity, and response time behavior. Of the fibers tested, that coated with 75 microm Carboxen/poly(dimethylsiloxane) was found to be an excellent passive sampler for VOCs. TWA passive sampling with a SPME device was shown to be almost independent of face velocity and to be more tolerant of high and low analyte concentrations and long and short sampling times, because of the ease with which the diffusion path length could be changed. It was found that environmental conditions, e.g., temperature, pressure, relative humidity, and ozone, have little or no effect on sampling. The 75-microm Carboxen/poly(dimethylsiloxane) fiber can retain VOCs for up to two weeks without significant loss. When the SPME device was tested in the field and the results were compared with those from National Institute of Occupational Health and Safety method 1501, good agreement was obtained. PMID- 12720334 TI - Slurry sampling-microwave assisted leaching prior to hydride generation pervaporation-atomic fluorescence detection for the determination of extractable arsenic in soil. AB - A flow injection-pervaporation method, where the sample was introduced as slurry, has been developed for the continuous derivatization and determination of arsenic in soil by hydride generation-atomic fluorescence spectrometry. The removal of arsenic is achieved with the help of a microwave digestor, which facilitates an on-line leaching in the flow injection manifold. Slurries, prepared by mixing the soil (particle size <65 microm) with 6 mol L(-)(1) HCl, were magnetically stirred for 3 min, and while stirring, the pump aspirated the aliquot and filled the loop (500 microL) of the injection valve. An industrial soil and five types of soil (sandy, clayey, slimy, limy, organic) were selected for the optimization of the leaching and determination steps of arsenic, respectively. The results obtained from three certified reference materials [stream sediment GBW 07311 (188 microg/mL As), river sediment CRM 320 (76.7 microg/mL As), and soil GBW 07405 (412 microg/mL As)] using direct calibration against aqueous standards demonstrate the reliability of the method. The relative standard deviation for within-laboratory reproducibility was 4.5%. PMID- 12720335 TI - Robust classifier for the automated detection of ammonia in heated plumes by passive fourier transform infrared spectrometry. AB - An automated classification algorithm is implemented for the detection of ammonia vapor in heated plumes by passive Fourier transform infrared (FT-IR) spectrometry. This classification methodology allows the real-time detection of chemical signatures in gaseous effluents such as those generated from industrial processes. The characteristics of real-time implementation and excellent robustness are achieved by an analysis strategy based on the application of band pass digital filters to short segments of the interferogram data collected by the FT-IR spectrometer, followed by the use of piecewise linear discriminant analysis to obtain a yes/no classification regarding the presence of the analyte signature in the filtered data. The optimal classifier developed through this work is based on only 110 interferogram points and employs a single band-pass filter centered at 945 cm(-)(1) with a pass-band full width at half-maximum of 93 cm(-)(1). The average stop-band attenuation of the optimal filter is 42.1 dB. The robustness of the algorithm is tested by exposing it to chemical releases of sulfur hexafluoride, ethanol, methanol, sulfur dioxide, and hydrogen chloride that were not included in the development of the classifier. Excellent classification performance is demonstrated, with missed ammonia detections occurring at a rate of approximately 1%. The occurrence of false detections is less than 0.1% for SF(6) and less than 0.02% for the other interferences tested. PMID- 12720337 TI - A method for the detection of hydrocarbon pollution in soils by headspace mass spectrometry and pattern recognition techniques. AB - In the present work, we report a methodology for the rapid detection of soil pollution by hydrocarbons that is based on direct coupling of a headspace sampler with a mass spectrometer. With no prior treatment, the samples are subjected to the headspace generation process and the volatiles generated are introduced directly into the mass spectrometer, thereby obtaining a fingerprint of the sample analyzed. The mass spectrum corresponding to the mass/charge ratios (m/z) ranging between 49 and 160 atomic mass units (amu) contains the information related to the composition of the headspace and is used as the analytical signal for the characterization of the samples. Chemometric treatments, such as hierarchical cluster analysis (HCA), linear discriminant analysis (LDA), and soft independent modeling class analogy (SIMCA) were used to characterize the different types of samples analyzed. The main advantage of the proposed methodology is that no prior chromatographic separation and no sample manipulation are required. The method is rapid, simple, and in view of the results, highly suitable for detecting pollution in soils polluted by hydrocarbons. PMID- 12720336 TI - A LC/APCI-MS/MS method for analysis of imidacloprid in soils, in plants, and in pollens. AB - Imidacloprid, the most used systemic insecticide, is suspected of having harmful effects on honeybees at nanogram per bee or at microgram per kilogram levels. However, there is a lack of methodology to detect imidacloprid and its metabolites at such low levels. We developed a method for the determination of low amounts of imidacloprid in soils, plants (leaves and flowers), and pollens by using HPLC coupled to tandem mass spectrometry (APCI-MS/MS). Extraction, separation, and detection were performed according to quality assurance criteria, to Good Laboratory Practice, and to criteria from the directive 96/23/EC, which is designed for banned substances. The linear range of application is 0.5-20 microg/kg imidacloprid in soils, in plants, and in pollens, with a relative standard deviation of 2.9% at 1 microg/kg. The limits of detection and of quantification are LOD = 0.1 microg/kg and LOQ = 1 microg/kg, respectively. For the first time, this study permitted us to follow the fate of imidacloprid in the environment. When treated, flowers of sunflower and maize contain average values of approximately 10 microg/kg imidacloprid. This explains that pollens from these crops are contaminated at levels of a few micrograms per kilogram, suggesting probable deleterious effects on honeybees. PMID- 12720338 TI - Fluorescent peptide probes for high-throughput measurement of protein phosphatases. AB - A homogeneous microplate assay for the serine/threonine protein phosphatases PP1 and PP2A, employing fluorescent-labeled phosphopeptides, has been developed. Phosphopeptides derived from a phosphoacceptor site in myelin basic protein were designed with a cysteine adjacent to the phosphoresidue, allowing site-selective labeling with dyes. The fluorescence emission from the environmentally sensitive fluorophore 7-fluorobenz-2-oxa-1,3-diazole-4-sulfonamide was found to be sensitive to the phosphorylation status of an adjacent threonine residue. Upon complete dephosphorylation of the dye-labeled phosphopeptide, a 56% decrease in fluorescence intensity was observed. The change in fluorescence was correlated with the release of inorganic phosphate from the phosphopeptide as measured using the malachite green assay. Conjugation of the fluorophore to the phosphopeptide was found to have no adverse effect on catalysis. A series of four phosphopeptide substrates were developed and characterized to probe PP1 and PP2A activity. The optimum phosphopeptides were then used to determine inhibition parameters for three natural protein phosphatase inhibitors. The use of a peptide-based approach has introduced a degree of specificity not observed with many conventional phosphatase substrates, while retaining the advantages of a real-time homogeneous fluorescence-based format, making the assay ideal for high-density screening. PMID- 12720339 TI - Capillary electrophoresis micro X-ray fluorescence: a tool for benchtop elemental analysis. AB - A new tool was developed for separation and elemental detection by interfacing a simple capillary electrophoresis (CE) apparatus, constructed using a thin-walled fused-silica capillary, with a benchtop energy-dispersive micro X-ray fluorescence (MXRF) system. X-ray excitation and detection of the separated analytes was done using an EDAX Eagle II micro X-ray fluorescence system equipped with a polycapillary Rh target excitation source and a SiLi detector. It was demonstrated that this prototype system could be used for the separation and detection of species containing two different metals from one another, specifically Cu and Co. Free Co could also be separated from Co bound to cyanocobalamin (vitamin B-12). Two organic compounds were also separated from one another, a large biological protein, ferritin, from a small biological organic, cyanocobalamin. Preliminary average detection limits obtained on this system were on the order of 10(-)(4) M and compared favorably to those reported for the similar technique of CE-synchrotron XRF. CEMXRF allows for nondestructive, simultaneous, on-line, benchtop elemental analysis for chemical speciation applications. PMID- 12720340 TI - Voltammetric characterization of a N,N'-diphenyl-p-phenylenediamine-loaded screen printed electrode: a disposable sensor for hydrogen sulfide. AB - The voltammetric response of a 10% (by weight) N,N'-diphenyl-p-phenylenediamine (DPPD) and 90% (by weight) carbon and binder screen-printed electrode has been examined in aqueous media over a range of pH using cyclic voltammetry both in the presence and in the absence of sulfide. In the absence, the screen-printed electrode undergoes an initial oxidative process on the surface of the solid organic particles to form an insoluble layer of the corresponding cation radical salt, DPPD(*)(+)X(-), where X(-) is an anion present in the solution. The charge transfer is thought to occur at the three-phase boundary between solid DPPD, carbon, and the aqueous solution. At higher potentials, a second oxidative wave is observed that is attributed to the oxidation of the bulk DPPD with intercalation of the anion species present to form a solid phase of DPPD(*)(+)X( ). The two voltammetric processes were found to stabilize after repetitive scanning, after which time, sulfide was added to the solution. The voltammetric response was found to respond to sulfide by showing a decrease in both the oxidative and reductive waves, which can be attributed to the sulfide effectively blocking the three-phase boundary. The response was found to be independent of the electrode used and at pH 4 produced a linear range from 20 to 165 microM, and a limit of detection of 7.5 microM for sulfide detection was achieved. PMID- 12720342 TI - Numerical investigation of an electrochemically induced tagging in a nanospray for protein analysis. AB - An on-line tagging of target species is simulated using the finite element method. A numerical model of an electrochemical EC(2X)E mechanism in a flow channel cell has been developed, corresponding to the electrochemical generation of a probe and the subsequent homogeneous reaction with the target. The kinetic and convective aspects are validated on short electrode geometries before taking into account the depletion of the target species. The model is then assessed according to previous experimental results on the on-line tagging of proteins. The occurrence and the efficiency of the on-line tagging are studied for both pressure-driven and electroosmotic flows. The involved phenomena including kinetic aspects are described in detail. Finally, optimal conditions for an effective quantitative tagging are discussed. PMID- 12720341 TI - Coupling the lactate oxidase to electrodes by ionotropic gelation of biopolymer. AB - A direct ionotropic gelation of the polycationic biopolymer chitosan (CHIT) with the polyanionic enzyme lactate oxidase (LOx) was used to form thin biopolymer enzyme films on the surface of platinum electrodes. The electrochemical assays of such films revealed a well-defined capacity of CHIT for the retention of LOx. The stoichiometry of the CHIT-LOx polyelectrolyte complexes was found to be approximately 1:40, i.e., on average, 1 CHIT chain retained 40 molecules of LOx in the CHIT-LOx films. The enzyme retention was ascribed to strong electrostatic interactions between the LOx and a fraction of the protonated amino groups on the CHIT chains. Although the LOx is inherently unstable outside its natural matrix, it displayed high surface activity of 0.26 units cm(-)(2) in the matrix of CHIT. This correlated with good stability of the biopolymer-enzyme films as demonstrated by a constant response of Pt/CHIT-LOx electrodes to lactate during continuous 24-h testing. When compared to other single-film lactate sensors, the Pt/CHIT-LOx electrodes displayed the best combination of analytical properties in terms of a low detection limit (50 nM), high sensitivity (0.23 A M(-)(1) cm( )(2)), and fast response time (<1 s). Such a performance validated the CHIT-LOx system as an attractive sensing element for the development of new lactate biosensors. PMID- 12720343 TI - Carbon nanotube/teflon composite electrochemical sensors and biosensors. AB - The fabrication and attractive performance of carbon nanotube (CNT)/Teflon composite electrodes, based on the dispersion of CNT within a Teflon binder, are described. The resulting CNT/Teflon material brings new capabilities for electrochemical devices by combining the advantages of CNT and "bulk" composite electrodes. The electrocatalytic properties of CNT are not impaired by their association with the Teflon binder. The marked electrocatalytic activity toward hydrogen peroxide and NADH permits effective low-potential amperometric biosensing of glucose and ethanol, respectively, in connection with the incorporation of glucose oxidase and alcohol dehydrogenase/NAD(+) within the three-dimensional CNT/Teflon matrix. The accelerated electron transfer is coupled with minimization of surface fouling and surface renewability. These advantages of CNT-based composite devices are illustrated from comparison to their graphite/Teflon counterparts. The influence of the CNT loading upon the amperometric and voltammetric data, as well as the electrode resistance, is examined. SEM images offer insights into the nature of the CNT/Teflon surface. The preparation of CNT/Teflon composites overcomes a major obstacle for creating CNT-based biosensing devices and expands the scope of CNT-based electrochemical devices. PMID- 12720344 TI - Characterization of platinum nanoparticle-embedded carbon film electrode and its detection of hydrogen peroxide. AB - A method for the highly sensitive determination of acetylcholine (ACh) and choline (Ch) that employs a graphite-like carbon film electrode containing 6.5% platinum (Pt) nanoparticles was developed for use as a detector in microbore liquid chromatography (LC) with a postcolumn enzyme reactor. The film electrode was prepared by RF cosputtering carbon and Pt, which requires only a one-step formation process. This method can control the Pt content of the film at a relatively low deposition temperature (below 200 degrees C). The average size of the Pt nanoparticles was 2.5 nm. The film electrode showed excellent electrocatalytic activity, high sensitivity, and negligible baseline drift when detecting hydrogen peroxide. The electrode was modified with glucose oxidase and responded rapidly to glucose with a much more stable baseline current than at a Pt bulk electrode based sensor. Therefore, it is appropriate to employ the electrode to detect trace amounts of biomolecules, such as neurotransmitters and hormones combined with various oxidase enzymes. We used the electrode as a detector for microbore LC and observed a low detection limit of 2.5 and 2.3 fmol (10-microL injection) for ACh and Ch, respectively, which is approximately 1 order of magnitude lower than that of a Pt bulk electrode. PMID- 12720345 TI - Characteristic electrochemical responses of polymer microchannel-microelectrode chips. AB - Polymer microchannel chips (dimension of the channel, 20 microm (depth) x 100 microm (width)) integrated with dual microband electrodes were fabricated by an imprinting method, and their characteristic electrochemical responses were elucidated in detail. A silicon micromachined template fabricated by photolithography and dry etching techniques was used for imprinting. An aqueous solution of a ferrocene derivative was brought into the microchannel by pressure driven flow, and the electrode responses were studied on the basis of voltammetry and chronoamperometry. A linear sweep voltammetry of 1-hydroxyethylferrocene (FeCp-OH) in water demonstrated that the electrode responses in the microchannel chip were best characterized by one-dimensional diffusion along the channel length, reflecting the structural dimension of the channel. In generation collection mode experiments, furthermore, a collection efficiency as high as approximately 90% was attained in the microchannel owing to both restricted space and characteristics of solution flow in the channel. It was confirmed that diffusion and solution flow made molecular transport very efficient in the microchannel. The experimental results were also compared with those predicted by computer simulations. PMID- 12720346 TI - UV vapor generation for determination of selenium by heated quartz tube atomic absorption spectrometry. AB - A new vapor generation technique utilizing UV irradiation coupled with atomic absorption for the determination of selenium in aqueous solutions is described. In the presence of low molecular weight organic acid solutions, inorganic selenium(IV) is converted by UV irradiation to volatile selenium species, which are then rapidly transported to a heated quartz tube atomizer for detection by atomic absorption spectrometry. Optimum conditions for photochemical vapor generation and interferences from concomitant elements were investigated. Identification of the volatile products using cryotrapping GC/MS analysis revealed that inorganic selenium(IV) is converted to volatile selenium hydride, selenium carbonyl, dimethyl selenide, and diethyl selenide in the presence of formic, acetic, propionic, and malonic acids, respectively. In acetic acid solution, the efficiency of generation was estimated to be 50 +/- 10%. No interference from Ni(2+) and Co(2+) at concentrations of 500 and 100 mg L(-)(1), respectively, was evident. A detection limit of 2.5 microg L(-)(1) and a relative sensitivity of 1.2 microg L(-)(1) (1% absorption) with a precision of 1.2% (RSD, n = 11) at 50 microg L(-)(1) were obtained. PMID- 12720347 TI - Quantitative 1H NMR spectroscopy of blood plasma metabolites. AB - The absolute quantification of blood plasma metabolites by proton NMR spectroscopy is complicated by the presence of a baseline and broad resonances originating from serum macromolecules and lipoproteins. A method for spectral simplification of proton NMR spectra of blood plasma is presented. Serum macromolecules and metabolites are completely separated by utilizing the large difference in translational diffusion coefficients in combination with diffusion sensitized proton NMR spectroscopy. The concentration of blood plasma metabolites can be quantified by using formate as an internal concentration reference. The results are compared with those obtained with ultrafiltration, a traditional method for separating macromolecules and metabolites, and demonstrate an excellent correlation between the two methods. The general nature of diffusion sensitized NMR spectroscopy allows application on a wide range of biological fluids. PMID- 12720348 TI - Exploration of microwave plasma source cavity ring-down spectroscopy for elemental measurements. AB - We are exploring sensitive techniques for elemental measurements using cavity ring-down spectroscopy (CRDS) combined with a compact microwave plasma source as an atomic absorption cell. The research work marries the high sensitivity of CRDS with a low-power microwave plasma source to develop a new instrument that yields high sensitivity and capability for elemental measurements. CRDS can provide orders of magnitude improvement in sensitivity over conventional absorption techniques. Additional benefit is gained from a compact microwave plasma source that possesses the advantages of low power and low-plasma gas flow rate, which are of benefit for atomic absorption measurements. A laboratory CRDS system consisting of a tunable dye laser is used in this work for developing a scientific base and demonstrating the feasibility of the technique. A laboratory designed and -built sampling system for solution sample introduction is used for testing. The ring-down signals are monitored using a photomultiplier tube and recorded using a digital oscilloscope interfaced to a computer. Lead is chosen as a typical element for the system optimization and characterization. The effects of baseline noise from the plasma source are reported. A detection limit of 0.8 ppb (10(-)(10)) is obtained with such a device. PMID- 12720349 TI - Specific aptamer-protein interaction studied by atomic force microscopy. AB - Aptamers are a new class of synthetic DNA/RNA oligonucleotides generated from in vitro selection to selectively bind with various molecules. Due to their molecular recognition capability for proteins, aptamers are becoming promising reagents in protein detection and new drug development. In this study, the specific interaction between the protein immunoglobulin E (IgE) and its 37-nt aptamer has been measured directly by atomic force microscopy. The single molecule unbinding force between IgE and the aptamer is determined using the Poisson statistical method. The individual unbinding force between IgE and its monoclonal antibody has also been obtained and compared to that between IgE and the aptamer. The results reveal the high affinity of the aptamer to protein, which could match or even surpass that of the antibody to its antigen. PMID- 12720350 TI - Reactive polymer coatings: a first step toward surface engineering of microfluidic devices. AB - We report fabrication, characterization, and use of microfluidic analysis devices containing surface-immobilized cell-capturing molecules. Amino-terminated biotin ligands are immobilized onto the luminal surface of a microdevice and effectively support self-assembly of proteins, antibodies, and mammalian cells. For this purpose, chemical vapor deposition (CVD) polymerization is used to functionalize PDMS-made microfluidic devices with poly[para-xylylene carboxylic acid pentafluorophenolester-co-para-xylylene]. The resulting reactive coating shows excellent adhesion when deposited in thin films (approximately 100 nm, and the distribution of the pentafluorophenol ester groups is reasonably uniform within the microchannel inner surface, as examined by fluorescence microscopy. The utility of these devices for cell-based bioassays is demonstrated by monitoring the concentration-dependent effect of the disintegrin echistatin on cell adhesion. The described assay format could be relevant to clinical research in various fields, including angiogenesis research. PMID- 12720351 TI - Analysis of individual acidic organelles by capillary electrophoresis with laser induced fluorescence detection facilitated by the endocytosis of fluorescently labeled microspheres. AB - Submicrometer-sized fluorescent microspheres were loaded into the acidic organelles of NS-1 mouse myeloma cells via endocytosis. Confocal microscopy imaging showed that microspheres colocalized nearly perfectly with LysoTracker Red, a probe that stains acidic organelles. Unlike LysoTracker dyes that seem to leak from acidic organelles upon cell disruption, microspheres are retained within these organelles, facilitating their analysis following isolation. Using capillary electrophoresis (CE) with laser-induced fluorescence detection (LIF), the electrophoretic mobilities of acidic organelles were individually calculated and fluorescence intensities individually measured. When cells were incubated for sufficient time to allow for endocytosis (48 h) with 3.9 x 10(3) microspheres/cell, replicate CE-LIF analyses of the corresponding isolated fraction indicated a dramatic increase in the number of detected events (n = 1990 +/- 234) and in the overall fluorescence intensity of the individual events (0.38 +/- 0.01 RFU; average +/- SD; n = 3) over the corresponding <10-min incubations (n = 60; 0.21 RFU, respectively). In addition, a treatment with 4-fold increase in microsphere density (1.6 x 10(4) microspheres/cell), increased the number of detected individual events (n = 3427 +/- 101) and altered only slightly the fluorescence intensity and electrophoretic mobility distributions. The individual electrophoretic mobility values ranged from -1.45 x 10(-)(4) to -3.0 x 10(-)(4) cm(2) V(-)(1) s(-)(1) while the individual fluorescence values ranged from 0.1 V to over 8 V, demonstrating the benefit of detecting organelles individually rather than averaging their properties over single cells or bulk homogenates. PMID- 12720352 TI - Ion-pairing ability, chemical stability, and selectivity behavior of halogenated dodecacarborane cation exchangers in neutral carrier-based ion-selective electrodes. AB - Recently, it has been discovered that carba-closo-dodecaborates can be used as cation exchangers in neutral carrier-based ion-selective chemical sensors. Because of their inherent chemical stability and versatile functionalization chemistries, they offer many advantages that may potentially be exploited for ion analyses that require nontraditional sample conditions, including strongly acidic media. In this work, trimethylammonium salts of undecachlorinated (UCC), undecabrominated (UBC), hexabrominated (HBC), and undecaiodinated (UIC) carborane anions were prepared and evaluated for their potential use in solvent polymeric membrane-based sensors. Computational methods including Natural population analysis and electrostatic mapping were used to predict the ion-exchanging ability of each lipophilic anion. In addition, the sandwich membrane technique was used to evaluate the ion-pairing ability of each carborane anion in situ (i.e., within bis(2-ethylhexyl) sebacate (DOS)- and 2-nitrophenyl octyl ether (o NPOE)-plasticized ISE membranes). The results of the computational and potentiometric studies found that binding affinity of the anions followed the generalized trend HBC > UCC > UBC > UIC. PVC-DOS bulk optode thin films containing the chromoionophore ETH 5315 and a respective anion were used to determine the chemical stability/lipophilicity of the carboranes and tetrakis[3,5 bis(trifluoromethyl)phenyl] borate (TFPB) in acidic media (0.2 M HOAc) under flowing conditions. The studies found that in terms of stability/lipophilicity UIC > UBC > TFPB approximately UCC >> HBC. Electrodes containing a Pb(2+) selective ionophore, tert-butylcalix[4]arene-tetrakis(N,N dimethylthioacetamide)(lead IV), were used to evaluate the functionality of each cation exchanger. An evaluation of response characteristics such as slope and selectivity found that UIC and UBC were quite comparable to the behavior of TFPB. Interestingly, both UIC and UBC showed a marked selectivity improvement over cadmium, with log K(pot)(pb),(Cd) values of -7.19 and -7.29, respectively, with TFPB giving a value of -5.89. Demonstrating excellent stability and suitable electrostatic properties, the carboranes, UIC in particular, are a very promising alternative to the tetraphenylborates and should find widespread application in the field of chemical sensors. PMID- 12720353 TI - Applications of attenuated total reflection infrared spectroscopic imaging to pharmaceutical formulations. AB - This paper demonstrates an approach to obtain chemical images of pharmaceutical tablets using attenuated total reflection infrared (ATR-IR) spectroscopy. FT-IR images with different fields of view and spatial resolution have been obtained using a combination of different ATR accessories. FT-IR imaging with the diamond ATR accessory and micro-ATR imaging technique have been compared. With the diamond ATR imaging accessory, compaction to a tablet can be performed and the chemical image measured in situ. It has been found that the diamond ATR imaging accessory gives information on the overall distribution of different components in a tablet while the micro-ATR imaging technique provides a closer look at the tablet with 4-microm spatial resolution. Low-concentration components down to 0.5% have been detected by the micro-ATR method. Both experimental and commercial systems are studied in this paper. PMID- 12720354 TI - Detection and identification of single molecules in living cells using spectrally resolved fluorescence lifetime imaging microscopy. AB - The detection of single mRNA molecules tagged by microinjected, singly fluorescently labeled oligo(dT) 43-mer molecules in living cells in quasi-natural surrounding, that is, cell culture medium, is demonstrated. Single-stranded oligonucleotides were labeled at the 5'-end with a red-absorbing oxazine derivative (MR121) and excited by a pulsed laser diode emitting at 635 nm with a repetition rate of 64 MHz. Spectrally resolved fluorescence lifetime imaging microscopy (SFLIM) on untreated living 3T3 mouse fibroblast cells reveals autofluorescence signals found predominately in the cytoplasm with fluorescence lifetimes of approximately 1.3 ns and emission maximums of approximately 665-670 nm. Hence, fluorescence signals of single MR121-labeled oligonucleotide molecules that exhibit a fluorescence lifetime of 2.8 ns and a fluorescence emission maximum of 685 nm can be easily discriminated against autofluorescence. MR121 labeled oligonucleotides were microinjected into the cytoplasm or nucleus of living 3T3 mouse fibroblast cells using a micropipet. Since the micropipet exhibits an inner diameter of 500 +/- 200 nm at the very end of the tip comparable to the diameter of the detection volume applied-the number of molecules delivered into the cell via the micropipet can be counted. Furthermore, the presented technique enables the quantitative detection and time-resolved identification of single molecules in living cells as a result of their characteristic emission maximums and fluorescence lifetime. The results obtained from single-molecule studies demonstrate for the first time that 10-30% of the microinjected oligo(dT) 43-mer molecules cannot diffuse freely inside of the nucleus but, rather, are tethered to immobile elements of the transcriptional, splicing, or polyadenylation machinery. PMID- 12720355 TI - Scanning electrochemical microscopy-based drug sensitivity test for a cell culture integrated in silicon microstructures. AB - The respiratory activity of collagen-embedded living cells was imaged by scanning electrochemical microscopy (SECM) with the objective to study anticancer drug sensitivity. Two kinds of cancer cells, the human erythroleukemia cell line (K562) and its adriamycin-resistant subline (K562/ADM), were immobilized at the array of microholes micromachined on a silicon wafer for comparative characterization of their sensitivity to the anticancer drug, ADM. The results obtained by the SECM method showed correspondence to a conventional colorimetric assay (SDI assay). Furthermore, since the SECM assay is based on the noninvasive measurement of the respiration activity, continuous monitoring of a dose response was possible. PMID- 12720356 TI - Quantitative chemical proteomics for identifying candidate drug targets. AB - We have developed a systematic strategy for drug target identification. This consists of the following sequential steps: (1) enrichment of total binding proteins using two differential affinity matrixes upon which are immobilized positive and negative chemical structures for drug activity, respectively; (2) covalent labeling of the proteins with a new cleavable isotope-coded affinity tag (ICAT) reagent, followed by proteolysis of the combined proteins; (3) isolation, identification, and relative quantification of the tagged peptides by liquid chromatography-mass spectrometry; (4) array-based transcription profiling to select candidate proteins; and (5) confirmation of direct interaction between the activity-associated structure and the selected proteins by using surface plasmon resonance. We present a typical application to identify the primary binding protein of a novel class of anticancer agents exemplified by E7070. Our results suggest that this approach provides a new aspect of quantitative proteomics to find specific binding proteins from protein mixture and should be applicable to a wide variety of biologically active small molecules with unidentified target proteins. PMID- 12720357 TI - On-line monitoring of airborne chemistry in levitated nanodroplets: in situ synthesis and application of SERS-active Ag-Sols for trace analysis by FT-Raman spectroscopy. AB - We report a new strategy for on-line monitoring of chemical reactions in ultrasonically levitated, nanoliter-sized droplets by Raman spectroscopy. A flow through microdispenser connected to an automated flow injection system was used to dose picoliter droplets into the node of an ultrasonic trap. Taking advantage of the flow-through characteristics of the microdispenser and the versatility of the automated flow system, a well-defined sequence of reagents could be injected via the microdispenser into the levitated droplet placed in the focus of the collection optics of the Fourier transform Raman spectrometer. In that way, chemical reactions could be carried out and monitored on-line. The developed system was used for fast, reproducible, in situ synthesis of a highly active surface enhanced Raman scattering (SERS) sol resulting from the reduction of silver nitrate with hydroxylamine hydrochloride in basic conditions. With this chemical system, SERS substrate preparation could be achieved at room temperature and in short time. The in situ prepared silver sol was used for trace analysis of several organic test molecules that were injected into the levitated SERS-active droplet again using the microdispenser. The concentration dependence of the SERS spectra was studied using 9-aminoacridine, revealing that down to the femtogram region high-quality SERS spectra could be obtained. Additionally, SERS spectra of 6-mercaptopurine, thiamine, and acridine were recorded in the levitated drop as well. PMID- 12720358 TI - High-resolution field desorption/ionization fourier transform ion cyclotron resonance mass analysis of nonpolar molecules. AB - We report the first field desorption ionization broadband high-resolution (m/Deltam(50%) approximately 65 000) mass spectra. We have interfaced a field ionization/field desorption source to a home-built 9.4-T FT-ICR mass spectrometer. The instrumental configuration employs convenient sample introduction (in-source liquid injection) and external ion accumulation. We demonstrate the utility of this configuration by generating high-resolution positive-ion mass spectra of C(60) and a midboiling crude oil distillate. The latter contains species not accessible by common soft-ionization methods, for example, low-voltage electron ionization, electrospray ionization, and matrix assisted laser desorption/ionization. The present work demonstrates significant advantages of FI/FD FT-ICR MS for analysis of nonpolar molecules in complex mixtures. PMID- 12720359 TI - Airborne chemistry coupled to Raman spectroscopy. AB - In this paper, the use of airborne chemistry (acoustically levitated drops) in combination with Raman spectroscopy is explored. We report herein the first Raman studies of crystallization processes in levitated drops and the first demonstration of surface-enhanced Raman scattering (SERS) detection in this medium. Crystallization studies on the model compounds benzamide and indomethacin resulted in the formation of two crystal modifications for each compound, suggesting that this methodology may be useful for investigation of polymorphs. SERS detection resulted in a signal enhancement of 27 000 for benzoic acid and 11 000 for rhodamine 6-G. The preliminary results presented here clearly indicate that several important applications of the combination between Raman spectroscopy and acoustic drop levitation can be expected in the future. PMID- 12720360 TI - Electrochemical genosensor based on colloidal gold nanoparticles for the detection of Factor V Leiden mutation using disposable pencil graphite electrodes. AB - Electrochemical genosensors for the detection of the Factor V Leiden mutation from polymerase chain reaction (PCR) amplicons using the oxidation signal of colloidal gold (Au) is described. A pencil graphite electrode (PGE) modified with target DNA, when hybridized with complementary probes conjugated to Au nanoparticles, responded with the appearance of a Au oxide wave at approximately +1.20 V. Specific probes were immobilized onto the Au nanoparticles in two different modes: (a) Inosine-substituted probes were covalently attached from their amino groups at the 5' end using N-(3-dimethylamino)propyl)-N' ethylcarbodiimide hydrochloride (EDC) and N-hydroxysulfosuccinimide (NHS) as a coupling agent onto a carboxylate-terminated l-cysteine self-assembled monolayer (SAM) preformed on the Au nanoparticles, and (b) probes with a hexanethiol group at their 5' phosphate end formed a SAM on Au nanoparticles. The genosensor relies on the hybridization of the probes with their complementary targets, which are covalently immobilized at the PGE surface. Au-tagged 23-mer capture probes were challenged with the synthetic 23-mer target, 131-base single-stranded DNA or denatured 256-base polymerase chain reaction (PCR) amplicon. The appearance of the Au oxidation signal shortened the assay time and simplified the detection of the Factor V Leiden mutation from PCR amplified real samples. The discrimination between the homozygous and heterozygous mutations was also established by comparing the peak currents of the Au signals. Numerous factors affecting the hybridization and nonspecific binding events were optimized. The detection limit for the PCR amplicons was found to be as low as 0.78 fmol; thus, it is suitable for point-of-care applications. PMID- 12720361 TI - Capillary electrophoresis to mass spectrometry interface using a porous junction. AB - A new capillary electrophoresis interface to electrospray ionization mass spectrometry (CE/ESI-MS) is introduced in which the electrical connection to the CE capillary outlet/ESI electrode is achieved by transfer of small ions related to the background electrolyte (BGE) through a porous section near the CE capillary outlet. In this design, only a small section of the capillary wall is made porous. The porous section is created by first thinning a small section of the capillary wall by drilling a well into it and then etching the remaining thin wall porous. This design has two advantages over previous designs (in which the whole circumference of the capillary was made porous): first, the capillary interface is more robust because only a small section of it is made porous, and therefore, no liquid junction is needed to secure the porous section. The electrical connection is achieved simply by inserting the capillary outlet containing the porous junction into the existing ESI needle and filling the needle with the BGE. Second, the time required to make the fused silica porous is reduced from approximately 1 h to a few minutes. In addition, there is no dead volume associated with the porous design, and because the actual metal/liquid contact occurs outside of the CE capillary, bubble formation due to redox reactions of water at the electrode does not affect CE/ESI-MS performance. The performance of this interface is demonstrated by the analyses of peptide and protein mixtures. PMID- 12720362 TI - A high-throughput method for the conversion of CO2 obtained from biochemical samples to graphite in septa-sealed vials for quantification of 14C via accelerator mass spectrometry. AB - The growth of accelerator mass spectrometry as a tool for quantitative isotope ratio analysis in the biosciences necessitates high-throughput sample preparation. A method has been developed to convert CO(2) obtained from carbonaceous samples to solid graphite for highly sensitive and precise (14)C quantification. Septa-sealed vials are used along with commercially available disposable materials, eliminating sample cross contamination, minimizing complex handling, and keeping per sample costs low. Samples containing between 0.25 and 10 mg of total carbon can be reduced to graphite in approximately 4 h in routine operation. Approximately 150 samples per 8-h day can be prepared by a single technician. PMID- 12720364 TI - Mushroom tyrosinase: recent prospects. AB - Tyrosinase, also known as polyphenol oxidase, is a copper-containing enzyme, which is widely distributed in microorganisms, animals, and plants. Nowadays mushroom tyrosinase has become popular because it is readily available and useful in a number of applications. This work presents a study on the importance of tyrosinase, especially that derived from mushroom, and describes its biochemical character and inhibition and activation by the various chemicals obtained from natural and synthetic origins with its clinical and industrial importance in the recent prospects. PMID- 12720365 TI - Nonprotein amino acids of plants: significance in medicine, nutrition, and agriculture. AB - Those nonprotein amino acids found in food and fodder plants and known to be toxic to man and domestic animals are described. These include toxins from many legume genera including Lathyrus, from other higher plant families, from seaweeds, and from fungi. Some inhibit protein synthesis, while others are incorporated into proteins with toxic effects. Basic processes such as urea synthesis and neurotransmission may be disrupted. The probable roles of nonprotein amino acids in protecting plants against predators, pathogens, and competing plant species are considered. The need to learn more of the nutritive value of nontoxic nonprotein amino acids and to explore the potential of others either as drugs or as leads to drugs in human and veterinary medicine is emphasized. PMID- 12720367 TI - Direct fluorometric determination of fluorescent substances in powders: the case of riboflavin in cereal flours. AB - Front-face emission spectra of powders can be recorded with a commercial spectrofluorometer. By combining the emissions of a scatterer powder and of a wheat flour sample, the scattering contribution to the front-face emission spectra of flour is removed, and the fluorescence of the flour is isolated. The fluorescence depends on the concentration of the fluorophores. By choosing convenient measurement parameters and by measuring the emission spectra of flour samples suitably enriched with riboflavin, the fluorescence of riboflavin could be isolated from that of other substances present in flours and the concentration of vitamin B(2) in native substrates could be determined. This method is particularly apt for the measurement of vitamin B(2) in low riboflavin-containing powders such as wheat flours, which are usually analyzed through complex chemical and microbiological methods. The method is essentially phenomenological, in view of the interpretation difficulties connected to the origin of the fluorescence resulting from the absorption of multiply scattered photons. PMID- 12720366 TI - Phenolic acids in foods: an overview of analytical methodology. AB - Phenolic acids are aromatic secondary plant metabolites, widely spread throughout the plant kingdom. Existing analytical methods for phenolic acids originated from interest in their biological roles as secondary metabolites and from their roles in food quality and their organoleptic properties. Recent interest in phenolic acids stems from their potential protective role, through ingestion of fruits and vegetables, against oxidative damage diseases (coronary heart disease, stroke, and cancers). High performance liquid chromatography (HPLC) as well as gas chromatography (GC) are the two separation techniques reviewed. Extraction from plant matrixes and cleavage reactions through hydrolysis (acidic, basic, and enzymatic) are discussed as are the derivatization reagents used in sample preparation for GC. Detection systems discussed include UV-Vis spectroscopy, mass spectrometry, electrochemical, and fluorometric detection. The most common tandem techniques are HPLC/UV and GC/MS, yet LC/MS is becoming more common. The masses and MS fragmentation patterns of phenolic acids are discussed and tabulated as are the UV absorption maxima. PMID- 12720368 TI - Detection of phloridzin in strawberries (Fragaria x ananassa Duch.) by HPLC-PDA MS/MS and NMR spectroscopy. AB - The phenolic profile of strawberry fruits (Fragaria x ananassa Duch., Rosaceae) was investigated by high-performance liquid chromatography with photodiode array detection. A peak displaying retention time and UV spectral data identical to those of phloridzin (phloretin 2'-O-beta-d-glucoside), a dihydrochalcone glucoside so far considered characteristic of apples, was monitored. For further characterization, crude extracts of strawberries were purified on polyamide, and the target compound was isolated by preparative and analytical HPLC. Structure elucidation was performed on the basis of APCI- and ESI-MS in the negative ion mode as well as by 1D and 2D NMR spectroscopy using authentic phloridzin for comparison. The d-configuration of the sugar moiety was established by HPLC analysis of the corresponding acyclic 1-deoxy-1-(N-acetyl-alpha methylbenzylamino)alditol acetate. Apart from its chemotaxonomic relevance, this first report on the occurrence of phloridzin in strawberries is of particular interest for the authenticity control of strawberry products such as juices, jams, and fruit preparations since phloridzin has so far been used for the detection of fraudulent admixtures. PMID- 12720369 TI - Hierarchical scheme for LC-MSn identification of chlorogenic acids. AB - The fragmentation behavior of 18 chlorogenic acids that are not substituted at position 1 has been investigated using LC-MS(4) applied to a methanolic coffee bean extract and commercial cider (hard cider). Using LC-MS(3), it is possible to discriminate between each of the three isomers of p-coumaroylquinic acid, caffeoylquinic acid, feruloylquinic acid, and dicaffeoylquinic acid, and a hierarchical key has been prepared to facilitate this process when standards are not available. MS(4) fragmentations further support these assignments, but were not essential in reaching them. The distinctive behavior of 4-acyl and 3-acyl chlorogenic acids compared with the 5-acyl chlorogenic acids is a key factor permitting these assignments. The fragmentation patterns are dependent upon the particular stereochemical relationships between the individual substituents on the quinic acid moiety. Fragmentation is facilitated by 1,2-acyl participation and proceeds through quinic acid conformers in which the relevant substituents transiently adopt a 1,3-syn-diaxial relationship. Selected ion monitoring at m/z 529 clearly indicated the presence in coffee of six caffeoylferuloylquinic acid isomers, whereas previously only two or three had been demonstrated. The hierarchical key permitted specific structures to be assigned to each of the six isomers. These assignments are internally consistent and consistent with the limited data previously available. PMID- 12720370 TI - Determination of peroxyl radical scavenging activity of flavonoids and plant extracts using an automatic potentiometric titrator. AB - A novel potentiometric method for evaluation of peroxyl radical scavenging activity of flavonoids and plant extracts was developed. The oxidation of potassium iodide (KI) was performed in acetonitrilephosphate buffer (1:1) containing antioxidant using 2,2'-azobis(2-amidinopropane) dihydrochloride as a peroxyl radical generator. The amount of iodine released from KI during a 20-min free radical oxidation was determined quantitatively using an automatic potentiometric titrator with sodium thiosulfate. The radical scavenging activity of the sample was expressed as the inhibition ratio for iodine release of the control group mediated by the radical. The results obtained from some authentic polyphenols correlated well with those of previous reports. This is a simple, time-saving method requiring less than 30 min and is useful in assessing the radical scavenging activity of antioxidants in plant extracts. We describe the radical scavenging activities of various flavonoids including 21 kinds of tea catechins and vegetable extracts by this method. PMID- 12720371 TI - Acid detergent fiber analysis in oilseed Brassicas by near-infrared spectroscopy. AB - The potential of near infrared spectroscopy (NIRS) for determining the acid detergent fiber (ADF) in the seed of oilseed Brassica (fam. Brassicaceae) was assessed. One hundred and fifty accessions belonging to the species Indian mustard (Brassica juncea L. Czern.& Coss.), Ethiopian mustard (B. carinata A. Braun) and rapeseed (B. napus L.) were scanned by NIRS as intact and ground seed, and their ADF values were regressed against different spectra transformations by modified partial least squares regression. The coefficients of determination in the external validation (r(2)) for intact and ground seed were 0.83 and 0.85, respectively. The standard deviation to standard error of prediction ratio and range to standard error of prediction ratio were 2.40 and 10.75 for intact seed and 2.62 and 11.76 for ground seed. No significant differences in the prediction were found for both sample presentations. Effects of the C-H and O-H groups of lipids and water, respectively, as well as protein and chlorophyll, were most important in modeling these equations. PMID- 12720372 TI - Herbicidal, plant growth inhibitory, and cytotoxic activities of bismuthines containing aromatic heterocycles. AB - This work presents the herbicidal and plant growth regulatory activities of tertiary bismuthines containing heterocyclic aromatic rings of the general formula (2-C(4)H(3)X)(3)Bi, where X = S (3), O (1), or NMe (2). Toxicity against Artemia salina and herbicidal activity on Lactuca sativa, Trifolium pratense, and Lolium multiflorum were tested. In addition to the effects on mitochondrial respiration obtained from roots of Phaseolus vulgaris, these compounds also demonstrated partial radical scavenging properties against 2,2-diphenyl-1 picrylhydrazyl (DPPH). The furyl substituent is the most important structural requirement for the activity measurements observed in this study. PMID- 12720374 TI - 6beta-hydroxygedunin from Azadirachta indica. Its potentiation effects with some non-azadirachtin limonoids in neem against lepidopteran larvae. AB - The biological activity of 6beta-hydroxygedunin isolated from Azadirachta indica A. Juss. was assessed using the gram pod borer, Helicoverpa armigera (Hubner), and Asian armyworm, Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae), alone and in combination with other limonoids, gedunin, salannin, nimbinene, and azadirachtin. The compound exhibited growth inhibitory activity in artificial diet bioassays, with 24.2 and 21.5 ppm, respectively, inhibiting growth by 50%. This efficacy was higher in comparison to gedunin (EC(50) = 50.8 and 40.4 ppm), salannin (EC(50) = 74.5 and 72.0 ppm), and nimbinene (EC(50) = 391.4 and 404.5 ppm). Azadirachtin, however, remained the most active neem allelochemical against both insect species. Nutritional assays clearly demonstrated that, though relative consumption and growth rates of fourth instar larvae were reduced, gedunin-type compounds induced physiological toxicity, evident by reduced efficiency of conversion of ingested food (ECI) in feeding experiments. Salannin and nimbinene, on the contrary, induced concentration-dependent feeding deterrence only. In feeding experiments, combinations of the compounds revealed that when azadirachtin was present in a mixture, EC(50) values did not deviate from the individual efficacy of azadirachtin (0.26 and 0.21 ppm, respectively) against H. armigera and S. litura larvae. However, a combination without azadirachtin did show a potentiation effect with potent EC(50) values among structurally different molecules, i.e., when salannin or nimbinene was combined with 6beta-hydroxygedunin or gedunin rather than structurally similar salannin + nimbinene or 6beta-hydroxygedunin + gedunin. Obviously, azadirachtin being the most active compound in neem is not synergized or influenced by any other limonoid, but other non-azadirachtin limonoids were more potent in specific combinations vis-a-vis the structural chemistry of the compound. It is obvious from the present study that potentiation among non-azadirachtin limonoids having explicitly two different modes of action, such as feeding deterrence and physiological toxicity, may be playing a significant role in the potentiation effect. PMID- 12720373 TI - Antioxidant activity of dietary oregano essential oil and alpha-tocopheryl acetate supplementation in long-term frozen stored turkey meat. AB - The effects of dietary oregano essential oil and alpha-tocopheryl acetate supplementation on the oxidative stability of long-term frozen stored turkey meat were investigated. Thirty 12-week-old turkeys, randomly divided into five groups, were given a basal diet or a basal diet supplemented with 200 mg of alpha tocopheryl acetate kg(-1), or 100 or 200 mg of oregano oil kg(-1), or 100 mg of oregano oil plus 100 mg of alpha-tocopheryl acetate kg(-1) for 4 weeks prior to slaughter. Lipid oxidation in breast and thigh meat was assessed after 1, 3, 6, and 9 months of frozen storage at -20 degrees C prior to or following 7 days of refrigerated storage at 4 degrees C. Results showed that oregano oil increased the oxidative stability of breast and thigh meat during the frozen storage. Dietary oregano oil at the inclusion level of 200 mg kg(-1) feed was significantly (p < 0.05) more effective in delaying lipid oxidation compared to the level of 100 mg kg(-1), but equivalent to dietary alpha-tocopheryl acetate supplementation at 200 mg kg(-1), which in turn was inferior to dietary supplementation of 100 mg kg(-1) oregano essential oil plus 100 mg kg(-1) alpha tocopheryl acetate that was significantly (p < 0.05) superior to all other treatments. Thigh meat was more susceptible to oxidation than breast meat, although the former contained alpha-tocopherol at markedly higher levels. Mean alpha-tocopherol levels in breast and thigh meat from all treatments decreased during the frozen storage, the decrease being sharper between 1 and 3 months of frozen storage for breast and between 3 and 6 months for thigh meat. Oregano oil supplementation increased (p < 0.05) the retention of alpha-tocopherol in meat, the increase being positively correlated with the supplementation level. However, the retention of alpha-tocopherol in meat could only partly elucidate the antioxidant activity exhibited by dietary oregano oil supplementation. PMID- 12720375 TI - Quantity and potential biological activity of caffeic acid in sweet potato [Ipomoea batatas (L.) Lam.] storage root periderm. AB - The caffeic acid content of storage root periderm and cortex tissues of genetically diverse sweet potato [Ipomoea batatas (L.) Lam.] cultivars and breeding clones was quantified by high-performance liquid chromatography. Periderm caffeic acid content of the clones ranged from 0.008 to 7.97 mg/g dry weight, whereas the highest cortex content was 0.047 mg/g. Clones varied greatly in periderm caffeic acid content in all experiments, but there were also differences between experiments in content averaged for all clones. This indicates that periderm caffeic acid content is subject to genetic and environmental influences. Caffeic acid inhibited the growth of four sweet potato pathogenic fungi and germination of proso millet seeds in bioassays. Inhibitory activity in the bioassays suggests that high periderm caffeic acid levels contribute to the storage root defense chemistry of some sweet potato genotypes. PMID- 12720376 TI - Isolation, structural elucidation, and inhibitory effects of terpenoid and lipid constituents from sunflower pollen on Epstein-Barr virus early antigen induced by tumor promoter, TPA. AB - Eight fatty acid esters of triterpene alcohols (1-8), four free triterpene alcohols (9, 12, 17, and 18), four diterpene acids (19-22), two tocopherol related compounds (23 and 24), four estolides (25-28), three syn-alkane-4,6-diols (29-31), one 1,3-dioxoalkanoic acid (32), and one aliphatic ketone (33), along with the mixture of free fatty acids, were isolated from the diethyl ether extract of the pollen grains of sunflower (Helianthus annuus). Among these compounds, 14 (2-8, 12, 23, 25-28, and 33) were new naturally occurring compounds, and their structures were determined on the basis of spectroscopic methods. Twenty-four terpenoids and lipids (1-4, 6-9, 12, and 19-33) and six free triterpene triols (10, 11, and 13-16), derived from their fatty acid esters (2, 3, and 5-8) by alkaline hydrolysis, were evaluated with respect to their inhibitory effects on the induction of Epstein-Barr virus early antigen (EBV-EA) induced by the tumor promoter, 12-O-tetradecanoylphorbol-13-acetate (TPA), in Raji cells, which is known to be a primary screening test for antitumor promoters. Among the 30 compounds tested, 21 compounds possessing a di- or a polycyclic ring system in the molecule (1-4, 6-16, and 19-24) showed potent inhibitory effects on EBV-EA induction (91-100% inhibition at 1 x 10(3) mol ratio/TPA). PMID- 12720377 TI - Application of hexanal, E-2-hexenal, and hexyl acetate to improve the safety of fresh-sliced apples. AB - The aims of this work were to evaluate the effects of different concentrations of hexanal, (E)-2-hexenal, hexyl acetate, and their mixtures on the fate of pathogenic species such as Escherichia coli, Salmonella enteritidis, and Listeria monocytogenes inoculated in model systems as well as the antimicrobial activity against the target species of the chosen molecules when added to the packaging atmosphere of inoculated fresh-sliced apples. The result obtained in this work pointed out the potential use of compounds such as hexanal, (E)-2-hexenal, and hexyl acetate for both the extension of shelf life and an improvement of hygienic safety of "minimally processed foods". In fact, hexanal, (E)-2-hexenal, and hexyl acetate had a significant inhibitory effect against pathogen microorganisms frequently isolated from raw materials (E. coli, S. enteritidis, and L. monocytogenes) when inoculated in both model and real systems. In this last condition, these compounds, at the levels used (150, 150, and 20 ppm for hexanal, hexyl acetate, and (E)-2-hexenal, respectively), displayed a bactericide effect on L. monocytogenes and they exhibited significant extensions of lag phase of E. coli and S. enteritidis inoculated at levels of 10(4)-10(5) CFU/g. PMID- 12720378 TI - Glycoalkaloid and calystegine contents of eight potato cultivars. AB - Diverse procedures have been reported for the separation and analysis by HPLC of the two major glycoalkaloids present in potatoes, alpha-chaconine and alpha solanine. To further improve the usefulness of the HPLC method, studies were carried out on the influence of several salient parameters on the analysis of the two potato glycoalkaloids. Effects on retention (elution, separation) times of the (a) composition and pH of the mobile phase (acetonitrile and phosphate buffer), (b) concentration of the phosphate buffer, (c) capacity values of column packing of four commercial HPLC amino columns, (d) column temperature were studied. Except for pH, all of the variables significantly influenced the retention times. The results make it possible to select analysis conditions that produce well-separated as well as symmetrical peaks of the two glycoalkaloids. This improved HPLC method (limit of detection of approximately 150 ng) was evaluated with extracts from the cortex of one whole potato variety (May Queen) grown in Japan and the freeze-dried peel and flesh from the following eight cultivars grown in the United States: Atlantic, Dark Red Norland, Ranger Russet, Red Lasoda, Russet Burbank, Russet Norkota, Shepody, and Snowden. In addition, the same samples were analyzed by GC-MS for the presence of two water-soluble nortropane alkaloids, calystegine A(3) and calystegine B(2), reported to be potent glycosidase inhibitors. The following ranges for the eight varieties of total glycoalkaloid and calystegine levels were observed: dry flesh, 5-592 and 6 316 mg/kg; dry peel, 84-2226 and 218-2581 mg/kg; dry whole potatoes, 40-883 and 34-326 mg/kg; wet flesh, 1-148 and 1-68 mg/kg; wet peel, 12-429 and 35-467 mg/kg; wet whole potatoes, 7-187 and 5-68 mg/kg. The possible significance of the results to plant and food sciences is discussed. PMID- 12720379 TI - Antitumor principle constituents of Myrica rubra Var. acuminata. AB - Myrica rubra var. acuminata is a native shrub widely distributed and used as folk medicine in Taiwan for stomach disorders and diarrhea. Column chromatography combined with cytotoxic bioassay-guided fractionation was performed to isolate the antitumor principles from fresh leaves of M. rubravar. acuminata. The 20% MeOH eluate fraction of M. rubra var. acuminata inhibited the viability of HeLa and P-388 cells in an in vitro assay and an in vivo P-388 tumor-bearing CDF(1) mouse model. The percent increase in life span (%ILS) of 20% MeOH eluate fraction was greater than 125%. (-)-Epigallocatechin 3-O-gallate (1) and prodelphinidin A 2,3'-O-gallate (2) were isolated from D-20 as the antitumor principle components. Both compounds can inhibit the growth of HeLa cells, but 1 had lower cytotoxic effects in normal cervical fibroblasts than did 2. Moreover, pretreatment with a caspase-3 specific inhibitor prevented 1- and 2-induced poly(ADP-ribose) polymerase cleavage. In view of these results, we suggest that 1 and 2 can induce apoptosis in HeLa cells and that activation of caspase-3 may provide a mechanistic explanation for their cytotoxic effects. Therefore, we suggest that the 20% MeOH eluate fraction extract is good for health and that M. rubra var. acuminata is an economically valuable plant. PMID- 12720380 TI - Purification and characterization of an N-acetylglucosamine-binding lectin from Koelreuteria paniculata seeds and its effect on the larval development of Callosobruchus maculatus (Coleoptera: Bruchidae) and Anagasta kuehniella (Lepidoptera: Pyralidae). AB - This study describes the purification of an N-acetylglucosamine-binding lectin from Koelreuteria paniculata seeds and its effects on the larval development of Callobruchus maculatus and Anagasta kuehniella. The lectin (KpLec) was characterized and isolated by gel filtration, affinity column, and reverse phase chromatography. SDS-PAGE indicated that this lectin is a dimer composed of subunits of 22 and 44 kDa. The N terminus exhibited 40% similarity with Urtiga dioica agglutinin. KpLec was tested for anti-insect activity against C. maculatus and A. kuehniella. With regard to C. maculatus, an artificial diet containing 0.7 and 1% KpLec produced LD(50) and ED(50) value, respectively. However, for A. kuenhiella, an artificial diet containing 0.65% KpLec produced an LD(50), whereas 0.2% KpLec produced an ED(50). The transformation of genes coding for this lectin could be useful in the development of insect resistance in important agricultural crops. PMID- 12720381 TI - Sex pheromone of the oak processionary moth Thaumetopoea processionea. Identification and biological activity. AB - The sex pheromone of the oak processionary moth Thaumetopoea processionea has been characterized from female gland extracts as a mixture of (Z,Z)-11,13 hexadecadienyl acetate (1), (E,Z)-11,13-hexadecadienyl acetate (3) and (Z,Z) 11,13-hexadecadienol (2) in 88:7:5 ratio. The amount of the major compound 1 was 20-30 ng/gland. No trace of (Z,Z)-11,13-hexadecadienal was found in the extract, and therefore, T. processionea appears to be the only "summer" processionary moth lacking this compound as a pheromone compound. The alcohol 2 had also been previously found but is electrophysiologically inactive, and in wind tunnel assays it lowers the number of contacts with the source when mixed with the major compound 1. The major component 1 elicited males to display the complete behavioral sequence, but the amount of chemical needed was unexpectedly high in comparison to the activity displayed by virgin females and gland extracts. (E,E) 11,13-hexadecadienyl acetate (5) inhibits the attractant activity of the major component 1 when mixed with 1 in 1:10 and 1:1 ratios. The main constituent 1 is active in the field, but its tendency to isomerize into the corresponding E,E isomer (5) must be considered if effective formulations are to be prepared. PMID- 12720382 TI - Levels of active oxygen species are controlled by ascorbic acid and anthocyanin in Arabidopsis. AB - Stabilization of the levels of active oxygen species (AOS) is important to the survival of organisms. To clarify the system controlling levels of AOS in plants, this study used an electron spin resonance (ESR) method to directly measure superoxide radical (O(2)(.-)) scavenging activities in the wild-type Arabidopsis thaliana (Col and Ler ecotypes), two anthocyanin mutants (tt3 and ttg1), and an ascorbic acid mutant (vtc1). Under ordinary growth conditions, Arabidopsis contained superoxide-scavenging activity (SOSA) of approximately 300-500 SOD units/g of fresh weight. The ESR pattern indicated that most (40-50%) of this activity was due to ascorbic acid. For the analysis of SOSA under conditions of oxidative stress, synthesis of AOS was induced by gamma-irradiation. The radical scavenging activity in irradiated plants increased approximately 10-fold following an associated increase in the accumulation of ascorbic acid and anthocyanin. The accumulation of ascorbic acid and anthocyanin was suppressed by treatment with an antioxidant before irradiation and was induced by treatment with a radical-generating reagent. The contributions of ascorbic acid and anthocyanin to the total superoxide radical scavenging activity differed among ecotypes. In the Ler ecotype, ascorbic acid accumulated at twice the level of that in the Col ecotype, and induction of anthocyanin was half that in Col. To confirm the activity of ascorbic acid and anthocyanin against AOS stress, the viability of the wild type and mutants (tt2, tt3,tt5, ttg1, and vtc1) was examined after gamma-irradiation. Only the plants in which ascorbic acid and anthocyanin were induced had the ability to grow and flower. PMID- 12720383 TI - FUM13 encodes a short chain dehydrogenase/reductase required for C-3 carbonyl reduction during fumonisin biosynthesis in Gibberella moniliformis. AB - Fumonisins are polyketide-derived mycotoxins produced by the filamentous fungus Gibberella moniliformis (anamorph Fusarium verticillioides). Wild-type strains of the fungus produce predominantly four B-series fumonisins, designated FB(1), FB(2), FB(3), and FB(4). Recently, a cluster of 15 putative fumonisin biosynthetic genes (FUM) was described in G. moniliformis. We have now conducted a functional analysis of FUM13, a gene in the cluster that is predicted by amino acid sequence similarity to encode a short chain dehydrogenase/reductase (SDR). Mass spectrometric analysis of metabolites from FUM13 deletion mutants revealed that they produce approximately 10% of wild-type levels of B-series fumonisins as well as two previously uncharacterized compounds. NMR analysis revealed that the new compounds are similar in structure to FB(3) and FB(4) but that they have a carbonyl function rather than a hydroxyl function at carbon atom 3 (C-3). These results indicate that the FUM13 protein catalyzes the reduction of the C-3 carbonyl to a hydroxyl group and are the first biochemical evidence directly linking a FUM gene to a specific reaction during fumonisin biosynthesis. The production of low levels of FB(1), FB(2), FB(3), and FB(4), which have a C-3 hydroxyl, by the FUM13 mutants suggests that G. moniliformis has an additional C 3 carbonyl reductase activity but that this enzyme functions less efficiently than the FUM13 protein. PMID- 12720384 TI - Increased electron donor and electron acceptor characters enhance the adhesion between oil droplets and cells of Yarrowia lipolytica as evaluated by a new cytometric assay. AB - The adhesion of methyl ricinoleate droplets to cells of the yeast Yarrowia lipolytica was investigated. A new cytometric method, relying on the double staining of fatty globules with Nile Red and of cells with Calcofluor, enabled us to quantify methyl ricinoleate droplet adhesion to cells precultured on a hydrophilic or on a hydrophobic carbon source. In this last case, droplet adsorption was enhanced and a MATS (microbial adhesion to solvents) test revealed that this increase was due to Lewis acid-base interactions and not to an increase in the hydrophobic properties of the cell surface. These preliminary results demonstrate that the developed cytometric method is promising for various applications concerning the study of interactions between microorganisms and an emulsified hydrophobic substrates. PMID- 12720385 TI - Lead contamination in Portuguese red wines from the Douro region: from the vineyard to the final product. AB - To quantify lead contamination in wines and to try to identify major lead sources, two winemaking processes were followed during one annual cycle of wine production. Two vineyards from the Douro Portuguese region and two types of wine, one red table wine, which has been produced in a very modern winery, and one red fortified wine (similar to Port), which has been produced by a traditional vinification process, were selected for this study. Aerosols from the vineyards atmosphere, vineyard soil, vine leaves, grapes, and samples from the intermediary and final wine product were collected. Suitable pretreatments, namely, high pressure microwave assisted digestion (soil, leaves, and grapes) and UV irradiation (grape juices and samples from the different steps of the vinification processes), were used. The samples were analyzed in terms of lead total concentration and respective isotope ratios by using inductively coupled plasma mass spectrometry and atomic absorption spectrophotometry with electrothermal atomization. It was observed that the major sources of lead were in the vinification system, the more traditional one introducing more lead than the modern one. For the fortified wine, the lead concentration increased from 4.7 microg L(-1), in the grape juice, to 17.2 microg L(-1), in the final product, while for the table wine the increase was from 4.1 to 13.1 microg L(-)(1). Therefore, only about 1/4 (fortified wine) and 1/3 (table wine) of the lead total content of the final products came from soil and atmospheric deposition. Therefore, it is expected that marked reductions of the lead content in the wines would occur if the sources of lead were removed from the tubes and containers used in the vinification system, particularly by using welding alloys and small fittings free of lead. The lead levels in the vine leaves (global mean of 0.43 microg g(dry leave)(-1)) and grapes (global mean of 35 ng g(dry grape)(-1)) were similar in both vineyards. PMID- 12720386 TI - Influence of industrial processing on orange juice flavanone solubility and transformation to chalcones under gastrointestinal conditions. AB - Orange juice manufactured at industrial scale was subjected to digestion under in vitro gastrointestinal conditions (pH, temperature, and enzyme and chemical conditions) to evaluate the influence of individual industrial processing treatments on flavanone solubility, stability, and ability to permeate through a membrane under simulated physiological conditions. Four industrial processes including squeezing, standard pasteurization, concentration, and freezing were evaluated. Hand squeezing was compared with industrial squeezing. After in vitro gastrointestinal digestion of the orange juices, the flavanones able to permeate through a dialysis membrane, and those remaining in the retentate were evaluated by HPLC as were those present in the insoluble fraction. In all of the assayed orange juices, a high content of precipitated chalcones ( approximately 70% of the total flavanones) was formed under the physiological conditions of the gastrointestinal tract. Hand squeezing provided a higher concentration of flavanones in the permeated fraction and lower transformation to chalcones than industrial squeezing. Standard pasteurization did not influence the solubility and permeability of the orange juice flavanones and chalcones. Industrial concentration did not affect the amount of flavanones able to permeate but decreased the chalcones produced. Juices produced from frozen orange juice contained considerably smaller amounts of both soluble flavanones and insoluble chalcones. PMID- 12720387 TI - Health-promoting compounds in broccoli as influenced by refrigerated transport and retail sale period. AB - Total aliphatic and indole glucosinolates, phenolic compounds (flavonoids and hydroxycinnamoyl derivatives), and vitamin C contents were evaluated in freshly harvested broccoli (Brassica oleracea L., var. italica, cv. Marathon) inflorescences. These were film-wrapped and stored for 7 days at 1 degrees C to simulate a maximum period of commercial transport and distribution. After cold storage, inflorescences were kept for 3 days at 15 degrees C to simulate a retail sale period. For wrapping, low-density polyethylene (LDPE) of 11 microm thickness was used. Gas composition was about 17% O(2) and 2% CO(2) during cold storage and about 16% O(2) and 3-4% CO(2) during shelf life within packages. The predominant glucosinolates were 4-methylsulfinylbutyl-glucosinolate (glucoraphanin), 3 indolylmethyl-glucosinolate (glucobrassicin), and 1-methoxy-3-indolylmethyl glucosinolate (neoglucobrassicin). The predominant hydroxycinnamoyl derivatives were identified as 1,2,2'-trisinapoylgentiobiose, 1,2-diferuloylgentiobiose, 1,2' disinapoyl-2-feruloylgentiobiose, and 3-O-caffeoyl-quinic (neochlorogenic acid). Results showed major losses at the end of both periods, in comparison with broccoli at harvest. Thus, the respective losses, at the end of cold storage and retail periods, were 71-80% of total glucosinolates, 62-59% of total flavonoids, 51-44% of sinapic acid derivatives, and 73-74% caffeoyl-quinic acid derivatives. Slight differences in all compound concentrations between storage and retail sale periods were detected. Distribution and retail periods had minimal effects on vitamin C. Weight loss was monitored at the end of both periods. PMID- 12720388 TI - Dihydropiperazine neonicotinoid compounds. Synthesis and insecticidal activity. AB - Syntheses of various isomeric dihydropiperazines can be approached successfully by taking advantage of the regioselective monothionation of their respective diones. Preparation of the precursor unsymmetrical N-substituted piperazinediones from readily available diamines is key to this selectivity. The dihydropiperazine ring system, as exemplified in 1-[(6-chloropyridin-3-yl)methyl]-4-methyl-3 oxopiperazin-2-ylidenecyanamide (4) and 1-[(2-chloro-1,3-thiazol-5-yl)methyl]-4 methyl-3-oxopiperazin-2-ylidenecyanamide (25), has been shown to be a suitable bioisosteric replacement for the imidazolidine ring system contained in neonicotinoid compounds. However, placement of the cyanoimino electron withdrawing group further removed from the pyridine ring, as in 4-[(6 chloropyridin-3-yl)methyl]-3-oxopiperazin-2-ylidenecyanamide (3a), or relocation of the carbonyl group, as in 1-[(6-chloropyridin-3-yl)methyl]-4-methyl-5 oxopiperazin-2-ylidenecyanamide (5), results in significantly decreased bioisosterism. The dihydropiperazine ring system of 4 and 25 also lends a degree of rigidity to the molecule that is not offered by the inactive acyclic counterpart 2-[(6-chloropyridin-3-yl)-methyl-(methyl)amino]-2-(cyanoimino)-N,N dimethylacetamide (6). A pharmacophore model is proposed that qualitatively explains the results on the basis of good overlap of the key pharmacophore elements of 4 and imidacloprid (1); the less active regioisomers of 4 (3a, 5, and 6) feature a smaller degree of overlap. PMID- 12720389 TI - Influence of microbial inoculation (Pseudomonas sp. strain ADP), the enzyme atrazine chlorohydrolase, and vegetation on the degradation of atrazine and metolachlor in soil. AB - The concentrations of atrazine in the freshly added soils and the soils that had been incubated for 50 days significantly decreased 1 day after the addition of the enzyme atrazine chlorohydrolase or the soil bacterium Pseudomonas sp. strain ADP as compared with those in the uninoculated soils. Atrazine chlorohydrolase or ADP had no effect on the degradation of metolachlor. The half-lives of atrazine in the freshly added soils and in the aged soils after the treatment with atrazine chlorohydrolase or ADP markedly decreased as compared with those in the uninoculated soils. The half-lives of metolachlor in the aged soils were much longer than those of freshly added metolachlor. The percentage atrazine degraded in the freshly treated soils was much higher than that in the aged soils. This indicates that aging significantly decreased the bioavailability of atrazine. Vegetation significantly decreased the concentration of metolachlor. However, vegetation showed no effect on the degradation of atrazine. PMID- 12720391 TI - Is the negative charge on RNHSO3-M+ an essential requirement for sulfamate sweetness? AB - Although many structure-taste studies have been carried out on sulfamate (cyclamate) sweeteners, there are still some unanswered questions-notably whether the sulfamate anion, -NHSO(3)(-), is essential for sweetness in this class of compounds. The literature is contradictory on this point; therefore, 14 sulfamate esters RNHSO(3)R', which contain the sulfamate moiety but without the negative charge, i.e., -NHSO(3)(-), have been synthesized and tasted under standard conditions. Almost all of the esters were found to possess strong sweetness accompanied by bitterness. Because the esters had to be heated in water to 60 degrees C to dissolve them, it was necessary to check for partial hydrolysis to the free sulfamic acids, RNHSO(3)H, since they would be sweet and would invalidate the tasting results if formed. This was done by monitoring (gas-liquid chromatography) the formation of alcohol after heating. Negligible or very low hydrolysis to acid was found for all 14 esters. This work, in addition to answering an important structure-taste question, points the way to the potential use of suitable sulfamate esters as additives in situations where the more usual sodium sulfamate salts are unsuitable, for example, in hydrophobic media. PMID- 12720390 TI - Phytoene desaturase inhibition by O-(2-phenoxy)ethyl-N-aralkylcarbamates. AB - O-[1-Ethyl-2-(3-trifluoromethylphenoxy)]ethyl-N-benzylcarbamate exhibits a marked inhibition of carotenoid biosynthesis. Forty-one analogues were synthesized and assayed for plant-type phytoene desaturase (PDS) and zeta-carotene desaturase (ZDS) inhibition in a cell-free system using recombinant enzymes obtained from Escherichia coli transformants. The target enzyme of all carbamates synthesized in this study is PDS and not ZDS; no inhibition of ZDS was observed using a 10( 4) M inhibitor concentration. Four compounds, O-[1-ethyl-2-(3 trifluoromethylphenoxy)]ethyl-N-(2-phenylethyl)carbamate (23), O-[1-ethyl-2-(3 trifluoromethylphenoxy)]ethyl-N-(2-chlorobenzyl)carbamate (25), O-[1-ethyl-2-(3 trifluoromethylphenoxy)]ethyl-N-(2-chlorobenzyl)carbamate (26), and O-[1-methyl-2 (3-trifluoromethylphenoxy)]ethyl-N-benzylcarbamate (30), were the most potent PDS inhibitors. Their pI(50) values, the negative logarithms of the molar concentration that produces a 50% inhibition, were 7.5, representing the same inhibitory activity as norflurazon. With respect to a structure-activity relationship the oxygen atom of the phenoxy group and a carbamate structure in O (1-ethyl-2-phenoxy)ethyl-N-aralkylcarbamates studied were found to be essential for strong PDS inhibitors. Also, introduction of an ethyl group at the alpha position of the ethylene bridge between the phenoxy group and the carbamate was important for a strong PDS inhibitor. Substituents at the 2- and/or 3-position of the phenoxybenzene ring were found to be favorable to a strong PDS inhibition of the analogues. PMID- 12720392 TI - Volatile compounds of wines produced by cells immobilized on grape skins. AB - A biocatalyst was prepared by immobilization of Saccharomyces cerevisiae cells on grape skins. Repeated batch fermentations were conducted using the immobilized biocatalyst as well as the free yeast cells at 25, 20, 15, and 10 degrees C. The major volatile byproducts were determined by GC, whereas the minor volatile constituents were extracted in dichloromethane and analyzed by HRGC-MS. The qualitative profiles of the wines produced were similar in every case. Immobilized cells gave wines with higher contents of ethyl and acetate esters that increased with temperature decreases from 25 to 15 degrees C. The amount of volatile alcohols was more pronounced in wines produced by free cells and decreased dramatically at low fermentation temperatures (10 degrees C). PMID- 12720393 TI - Effects of hydrocolloid thickeners on the perception of savory flavors. AB - The perceived intensities of savory flavors in hydrocolloid-thickened solutions were investigated using sensory paired comparison tests between two distinct thickener concentrations (high and low viscosities). The perceived saltiness of 3.5 g/L NaCl was found to be significantly reduced (P < 0.01) at the higher thickener concentration of both hydroxypropylmethyl cellulose (HPMC) and lambda carrageenan, relative to the lower concentration. Mushroom flavor (8 ppm of 1 octen-3-ol with 3 g/L NaCl) was perceived as significantly more intense (P < 0.05) in 1.7 g/L lambda-carrageenan as compared with the same concentration of flavoring in 10.2 g/L lambda-carrageenan. Garlic flavor (2.5 ppm of diallyl disulfide with 2 g/L NaCl) was perceived to be significantly more intense in 2 g/L HPMC (P < 0.01) than in 10 g/L HPMC. However, when the NaCl concentration in the more viscous sample was increased to 3 g/L, the garlic flavor intensities of the two systems were not significantly different, suggesting a perceptual interaction (enhancement) between salt taste and garlic flavor. In vivo aroma release measurements from the same samples, using API-MS, showed that hydrocolloid concentration did not significantly alter the amount of mushroom or garlic aromas released when solutions were consumed. It was concluded that changes in perceived saltiness were driving the reduction in savory flavor perception even though the aroma stimulus was unchanged (a taste-aroma interaction). These findings parallel previous results in sweet hydrocolloid thickened solutions. PMID- 12720394 TI - Chemical composition of the essential oils of Juniperus from ripe and unripe berries and leaves and their antimicrobial activity. AB - The composition of the essential oil from ripe and unripe berries and leaves of Juniperus oxycedrus L. ssp. oxycedrus, Juniperus phoenicea ssp. turbinata and Juniperus communis ssp. communis was analyzed by GC-MS, and microbiological assays were carried out. Samples were collected in different localities (Sardinia, Italy) and hydro distilled. The yields ranged between 2.54% +/- 0.21 (v?w dried weight) and 0.04% +/- 0.00. A total of 36 components were identified. The major compounds in the essential oils were alpha-pinene, beta-pinene, delta-3 carene, sabinene, myrcene, beta-phellandrene, limonene, and D-germacrene. Both qualitative and quantitative differences between species and between different parts of the plant were observed. The essential oils and their major compounds were tested against Candida albicans, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa, and the minimum inhibitory concentration and minimum bactericidal concentration were determined. The results obtained led to a nonsignificant inhibitory effect, although all the essential oils from Juniperus phoenicea ssp. turbinata and the essential oil from leaves of Juniperus oxycedrus ssp. oxycedrus exhibited rather good or weak activity against Candida albicans and Staphylococcus aureus. PMID- 12720395 TI - Identification of odor-active 3-mercapto-3-methylbutyl acetate in volatile fraction of roasted coffee brew isolated by steam distillation under reduced pressure. AB - In a roasted Arabica coffee brew, the potent roasty odor quality compound was identified as 3-mercapto-3-methylbutyl acetate by comparison of its Kovats gas chromatography retention index, mass spectrum, and odor quality to those of the synthetic authentic compound. 3-Mercapto-3-methylbutyl acetate has been identified for the first time in the coffee, and according to the results of the aroma extract dilution analysis, the contribution of this compound to the flavor of the roasted coffee brew varied depending on the degree of the coffee bean roasting. The concentration of this compound in the coffee brews as with 3 mercapto-3-methylbutyl formate increased with an increase in the degree of roasting. However, the slope of the amount of both esters was different, and 3 mercapto-3-methylbutyl acetate hardly increased with a low degree of roasting at more than a 21 luminosity (L)-value, but it rapidly increased when the roasting degree of the coffee beans reached the L-value of 18. These results suggested that the contribution of 3-mercapto-3-methylbutyl acetate to the overall flavor is peculiar to the flavor of the highly roasted coffee. PMID- 12720396 TI - Changes in physicochemical characteristics and volatile constituents of yellow- and white-fleshed nectarines during maturation and artificial ripening. AB - Changes in the volatile composition during maturation and artificial ripening of yellow-fleshed nectarines were investigated by means of GC-FID and GC-MS over three years (1999-2001). Unripe and commercially ripe fruits were placed in ripening chambers until complete ripening and compared to tree-ripe nectarines. Firmness, weight, soluble solids (SS), titratable acidity (TA), SS/TA ratio, organic acids, and sugars were also determined. Furthermore, the tree-ripe and artificially ripened yellow-fleshed nectarines were subjected to sensory analysis by a taste panel. In 2001, a similar experiment on white-fleshed nectarines (cv. Vermeil) was performed to compare with the results obtained on yellow-fleshed nectarines. Levels of volatiles compounds, in particular, lactones and C(13) norisoprenoids, were found to be the same or higher in the artificially ripened nectarines compared with the tree-ripe nectarines. In addition, no significant difference was observed for sweetness, sourness, and the intensities of "peach odors" or "peach aroma" in the sensory analysis between the tree-ripe samples and the artificially ripened nectarines. However, in the latter the levels of SS and the SS/TA ratio were very close to those observed in the initial unripe samples and significantly lower than those observed in tree-ripe nectarines. PMID- 12720397 TI - The elusiveness of coffee aroma: new insights from a non-empirical approach. AB - Aroma is central to a pleasurable eating/drinking experience but is one of the most labile components of food. Coffee is an outstanding example. Attempts to avoid or control aroma degradation are often frustrated by ignorance of the microscopic mechanisms that are responsible for it. One of the processes most frequently invoked is radical formation, yet the identity of the radicals and their involvement in aroma degradation are poorly understood at the molecular level. Here a step forward in the fundamental understanding of this complex problem is taken by identifying the most relevant radicals and their products using first-principles calculations. Over 100 radicals originating from key aroma compounds found in coffee and other foods have been studied and classified according to an unambiguous criterion: their thermodynamic stability relative to common radical sources. This classification scheme predicts that most aroma molecules are resistant to both peroxidation and attack from phenolic antioxidants but are unstable with respect to radicals such as .OH. Dimers- generated from radical reactions--were also considered, and the most volatile species, which may further contribute to coffee aroma degradation, were focused on. Those--which are very few indeed--that have this potential have been identified. PMID- 12720398 TI - GC-olfactometric characterization of aroma volatiles from the thermal degradation of thiamin in model orange juice. AB - Model orange juice solutions containing 0.024 mM thiamin hydrochloride were stored for up to 8 weeks at 35 degrees C in amber glass containers. Volatiles were evaluated, primarily, using gas chromatography (GC) with olfactometry but also with flame ionization detector, pulsed-flame photometer detector (PFPD) (sulfur specific), and MS detection. Both 2-methyl-3-furanthiol (MFT) and its dimer, bis(2-methyl-3-furyl) disulfide (MFT-MFT) were identified thus confirming that thiamin could serve as the precursor to these potent off-flavors in thermally degraded citrus juices. Thirteen aroma active components were observed. MFT and MFT-MFT were observed after only a few days storage, and produced 33% of the total aroma activity after 7 d storage. Both compounds were observed olfactometrically earlier than they could be detected using PFPD. Other aroma active compounds included 4,5-dimethylthiazole (skunky, earthy), 3-thiophenethiol (meaty, cooked), 2-methyl-4,5-dihydro-3(2H)-thiophenone (sour-fruity, musty, green), 2-acethylthiophene (burnt), 2-formyl-5-methylthiophene (meaty), and 2 methyl-3-(methyldithio) furan (meaty). PMID- 12720399 TI - Chemoenzymatic synthesis of aroma active 5,6-dihydro- and tetrahydropyrazines from aliphatic acyloins produced by baker's yeast. AB - Twenty-five acyloins were generated by biotransformation of aliphatic aldehydes and 2-ketocarboxylic acids using whole cells of baker's yeast as catalyst. Six of these acyloins were synthesized and tentatively characterized for the first time. Subsequent chemical reaction with 1,2-propanediamine under mild conditions resulted in the formation of thirteen 5,6-dihydropyrazines and six tetrahydropyrazines. Their odor qualities were evaluated, and their odor thresholds were estimated. Among these pyrazine derivatives, 2-ethyl-3,5-dimethyl 5,6-dihydropyrazine (roasted, nutty, 0.002 ng/L air), 2,3-diethyl-5-methyl-5,6 dihydropyrazine (roasted, 0.004 ng/L air), and 2-ethyl-3,5 dimethyltetrahydropyrazine (bread crustlike, 1.9 ng/L air) were the most intensive-smelling aroma active compounds. PMID- 12720400 TI - Characterization of elemental composition in kiwifruit grown in northern Iran. AB - Kiwifruit (Actinidia chinensis) has different varieties such as Hayward, Bruno, Monty, and Abbott. These varieties are different in taste, odor, shape, and some chemical compositions. This work reports the elemental analysis of Cu, Fe, Mn, Zn, Ca, Mg, K, and Na in the different varieties of Hayward, Bruno, Monty, and Abbott grown in northern Iran. Results from elemental analysis were determined using atomic absorption and emission spectroscopy. Distributions and amounts of metals for different varieties of kiwifruit have been investigated. Results have shown that the presence of elements varied with different varieties of kiwifruit. The soils in which the kiwifruits have been cultivated also have been analyzed. PMID- 12720401 TI - Aqueous extracts from some muscles inhibit hemoglobin-mediated oxidation of cod muscle membrane lipids. AB - It was evaluated whether trout hemoglobin (Hb)-mediated oxidation of minced washed cod muscle lipids could be prevented by an aqueous isolate from cod and some other muscle sources. Lipid hydroperoxides and painty odor developed approximately 4 days faster in washed than unwashed cod mince. When adding back an aqueous fraction (press juice) isolated from unwashed mince to washed mince at 2-6-fold dilutions, development of hydroperoxides and painty odor was either delayed or completely prevented. The inhibitory substances were heat stable, and their effect was slightly reduced at reduced pH. The <1 kDa fractions of whole and heated press juices were as inhibitory as the unfractionated press juices. Inhibition by the unheated, heated, and ultrafiltered (30 kDa) press juices was lost after dialysis. These findings implied the presence of one or more highly effective aqueous low molecular weight antioxidants in cod muscle press juice. The same antioxidative properties were found in heated haddock, dab, and winter flounder muscle press juices but not in heated herring and chicken muscle press juices. Unheated chicken press juice was however highly inhibitory. PMID- 12720402 TI - Modified arabinoxylan-based films. Part B. Grafting of omega-3 fatty acids by oxygen plasma and electron beam irradiation. AB - Arabinoxylans (AXs) are byproducts of the cereal milling industry. To obtain high value products, AXs have been used as a film-forming agent. Hence, AX-based films are poor water vapor barriers. The objectives of this study were to graft omega-3 (omega3) fatty acids onto AX polymeric chains by using two new technologies: cold plasma and electron beam (EB) irradiation. Results show that the surface hydrophobicity of the modified films is higher than that of a waxy coating or a low-density polyethylene (LDPE) film. In addition, FTIR spectroscopy analysis reveals vibration bands attributed to new chemical functions. Finally, a decrease in water vapor permeability (WVP) is obtained for the film treated with the alpha linolenic acid-rich oil. This result could be explained by a better diffusion of this shorter polyunsaturated fatty acid into the AX network. Linseed oils provide better barrier properties and a higher surface hydrophobicity than oils extract from marine oils. Edible oils were chosen for edible application of these films to retard moisture transfers in stuffed biscuits. PMID- 12720403 TI - Isolation and partial characterization of rennet-like proteases from Australian cardoon (Cynara cardunculus L.). AB - The yield, protein content, proteolytic activity, and substrate specificity of crude and partially purified extracts from dried and fresh Australian cardoon (Cynara cardunculus L.) flowers were determined. Crude water extracts had high yield but low protein content and proteolytic activity, whereas citric acid extracts had low yield but high protein content and proteolytic activity. Fresh flower extracts gave higher yield and proteolytic activity but lower protein content in comparison with dried flower extracts. Purification with ammonium sulfate resulted in significantly increased proteolytic activity for water extracts from both fresh and dried cardoon flowers, whereas the proteolytic activity of citric acid extracts did not change significantly after purification. Irrespective of extraction method, all extracts had higher proteolytic activity against ovine whole and kappa-caseins compared to their bovine counterparts, showing optimal activity at 37 degrees C and pH 6.0. Separation of purified extracts by ion-exchange liquid chromatography yielded three active fractions, each of which when assayed with sodium dodecyl sulfate capillary electrophoresis revealed two subunits with molecular masses of 15.5 and 33.1 kDa, respectively. PMID- 12720404 TI - O/W emulsification for the self-aggregation and nanoparticle formation of linoleic acid-modified chitosan in the aqueous system. AB - Chitosan was modified by coupling with linoleic acid through the 1-ethyl-3-(3 dimethylaminopropyl)carbodiimide-mediated reaction to increase its amphipathicity for improved emulsification. The micelle formation of linoleic acid-modified chitosan in the 0.1 M acetic acid solution was enhanced by O/W emulsification with methylene chloride, an oil phase. The fluorescence spectra indicate that without emulsification the self-aggregation of LA-chitosan occurred at the concentration of 1.0 g/L or above, and with emulsification, self-aggregation was greatly enhanced followed by a stable micelle formation at 2.0 g/L. The addition of 1 M sodium chloride promoted the self-aggregation of LA-chitosan molecules both with and without emulsification. The micelles of LA-chitosan formed nanosize particles ranging from 200 to 600 nm. The LA-chitosan nanoparticles encapsulated the lipid soluble model compound, retinal acetate, with 50% efficiency. PMID- 12720405 TI - A novel convenient process to obtain a raw decaffeinated tea polyphenol fraction using a lignocellulose column. AB - Lignocellulose prepared from sawdust was investigated for its potential application in obtaining a raw decaffeinated tea polyphenol fraction from tea extract. Tea polyphenols having gallate residues, namely, (-)epigallocatechin gallate (EGCg) and (-)epicatechin gallate (ECg), were adsorbed on the lignocellulose column, while caffeine was passed through it. Adsorbed polyphenols were eluted with 60% ethanol, and the elute was found to consist mainly of EGCg and ECg. The caffeine/EGCg ratio was 0.696 before lignocellulose column treatment, but it became 0.004 after the column treatment. These results suggest that the lignocellulose column provides a useful and convenient process of purification of tea polyphenol fraction accompanied by decaffeination. PMID- 12720406 TI - Formation of conjugated linoleic acids in soybean oil during hydrogenation with a nickel catalyst as affected by sulfur addition. AB - The effects of sulfur addition on the formation of conjugated linoleic acid (CLA) isomers were studied during the hydrogenation of soybean oil with a nonselective type nickel catalyst. Sulfur addition greatly promoted CLA formation in soybean oil during hydrogenation. As the amount of sulfur increased to a certain level, the maximal quantity of CLA in soybean oil during hydrogenation increased greatly. However, further increase in sulfur addition above the certain level decreased CLA formation. The optimal sulfur level for the promotion of CLA formation differed greatly with the amount of nickel used. It was of great interest to find that the optimal ratio of sulfur to nickel for the promotion of CLA formation was always 0.06:1, regardless of the nickel amount used. At the same ratio of sulfur to nickel, higher nickel content induced significantly higher production of CLA (p < 0.05). At the optimal sulfur to nickel ratio, an increase in the nickel amount from 0.05 to 0.15% produced approximately 1.5 times higher levels of CLA during hydrogenation under the tested conditions. The CLA isomer compositions were greatly affected by both sulfur addition and amounts of nickel used for treatment. This is the first report of the possibility that the total quantity of CLA and their isomer composition could be manipulated during hydrogenation by controlling the amounts of sulfur and nickel. PMID- 12720407 TI - Cold-set globular protein gels: interactions, structure and rheology as a function of protein concentration. AB - We identified the contribution of covalent and noncovalent interactions to the scaling behavior of the structural and rheological properties in a cold gelling protein system. The system we studied consisted of two types of whey protein aggregates, equal in size but different in the amount of accessible thiol groups at the surface of the aggregates. Analysis of the structural characteristics of acid-induced gels of both thiol-blocked and unmodified whey protein aggregates yielded a fractal dimension (2.3 +/- 0.1), which is in line with other comparable protein networks. However, application of known fractal scaling equations to our rheological data yielded ambiguous results. It is suggested that acid-induced cold-gelation probably starts off as a fractal process, but is rapidly taken over by another mechanism at larger length scales (>100 nm). In addition, indications were found for disulfide cross-link-dependent structural rearrangements at smaller length scales (<100 nm). PMID- 12720408 TI - Identification of reaction products of acylated anthocyanins from red radish with peroxyl radicals. AB - Red radish anthocyanin extract, which consists of 12 known acylated anthocyanins, was reacted with 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) to generate peroxyl radicals under acidic pH conditions at 37 degrees C. The reaction products were isolated using preparative HPLC, and their chemical structures were determined to be p-hydroxybenzoic acid (1), 6-O-(E)-p-coumaroyl-2-O-beta-d- glucopyranosyl-alpha-d-glucopyranoside (3), p-coumaric acid (4), 6-O-(E)-feruloyl 2-O-beta-d-glucopyranosyl-alpha-d-glucopyranoside (5), and ferulic acid (6). Some products were not identified. HPLC analyses of the mixture of acylated pelargonidin isolated from red radish and AAPH revealed that the acylated pelargonidins possess the radical scavenging ability on some common sites even if the characteristics of the intramolecular acyl units are different. Degradation rates of acylated pelargonidins and the formation rates of the resulting reaction products were found to be quite different. PMID- 12720409 TI - Thermal properties of partially hydrolyzed starch-glycerophosphatidylcholine complexes with various acyl chains. AB - Complexes of starch and monoacyl-sn-glycerophosphatidylcholine (GPC) containing various acyl (myristoyl, palmitoyl, and stearoyl) chains were subjected to hydrolysis with glucoamylase (EC 3.2.1.3). The enzyme hydrolyzed approximately 40% of starch control and 20-28% of starch-GPC complexes. Among the GPCs examined, 1- and 2-monomyristoyl-sn-GPC showed the highest resistance to enzyme hydrolysis, and the hydrolysis rate of starch-GPCs was greater with longer chains. Enzymatic hydrolysis strongly affected the thermal properties of the starch. After enzymatic hydrolysis of starch-GPC complexes for 24 h, their thermograms had broader peaks with lower enthalpies than the corresponding starch without enzyme; however, the starch-GPC complexes showed little change. The surface of starch-GPC granules was less eroded. These results showed that the increasing amount of starch-GPC complexes could be more resistant to hydrolysis. PMID- 12720410 TI - Purification and characterization of vanilla bean (Vanilla planifolia Andrews) beta-D-glucosidase. AB - Vanilla bean beta-D-glucosidase was purified to apparent homogeneity by successive anion exchange, hydrophobic interaction, and size-exclusion chromatography. The enzyme is a tetramer (201 kDa) made up of four identical subunits (50 kDa). The optimum pH was 6.5, and the optimum temperature was 40 degrees C at pH 7.0. K(m) values for p-nitrophenyl-beta-D-glucopyranoside and glucovanillin were 1.1 and 20.0 mM, respectively; V(max) values were 4.5 and 5.0 microkat.mg(-1). The beta-D-glucosidase was competitively inhibited by glucono delta-lactone and 1-deoxynojirimycin, with respective K(i) values of 670 and 152 microM, and not inhibited by 2 M glucose. The beta-D-glucosidase was not inhibited by N-ethylmaleimide and DTNB and fully inhibited by 1.5-2 M 2 mercaptoethanol and 1,4-dithiothreitol. The enzyme showed decreasing activity on p-nitrophenyl-beta-D-fucopyranoside, p-nitrophenyl-beta-D-glucopyranoside, p nitrophenyl-beta-D-galactopyranoside, and p-nitrophenyl-beta-D-xylopyranoside. The enzyme was also active on prunasin, esculin, and salicin and inactive on cellobiose, gentiobiose, amygdalin, phloridzin, indoxyl-beta-D-glucopyranoside, and quercetin-3-beta-D-glucopyranoside. PMID- 12720411 TI - Involvement of dehydroalanine and dehydrobutyrine in the addition of glutathione to nisin. AB - Nisin variants and fragments were reacted with glutathione, and the products of the reactions were analyzed by matrix-assisted laser desorption/ionization time of-flight mass spectrometry (MALDI-TOF MS) and liquid chromatography/mass spectrometry (LC-MS). Reactions between glutathione and either [Ala5]nisin or [Ala33]nisin resulted in products with two glutathione molecules conjugated to one nisin variant molecule. Only one glutathione molecule was added to [Ala5,Ala33]nisin. Fragmentation of the nisin molecule resulted in nisin 1-12, nisin 1-20, and nisin 1-32 fragments. Each fragment retained two dehydro residues, which subsequently underwent reaction with glutathione. The data indicated that the dehydroalanine residues of nisin are sites of addition for glutathione. Such addition renders the nisin molecule inactive. PMID- 12720412 TI - Phenolic composition of champagnes from Chardonnay and Pinot Noir vintages. AB - Nineteen phenolic compounds including hydroxybenzoic acids, hydroxycinnamic acids, flavonoids, phenolic alcohols, and phenolic aldehydes have been identified and quantified in two monovarietal champagnes, Chardonnay and Pinot Noir, by using a reverse-phase high-performance liquid chromatography (HPLC) system coupled with diode array detection. The identification of four hydroxycinnamic tartaric esters (caftaric, coutaric, fertaric, and 2-S-glutathionylcaftaric acids), two flavanonols (astilbin and engeletin), and some other compounds was confirmed by HPLC coupled with mass spectrometry. Caftaric acid and tyrosol were the major phenols. Hydroxybenzoic acids and flavonoids were present at low concentrations. The phenolic compositions of 2000 and 2001 Chardonnay and Pinot Noir vary quantitatively according to the year and the variety, but the chemical natures of the molecules are the same. The total phenolic content determined by colorimetric measurement ranges from 176 to 195 mg/L of gallic acid equivalent and is similar to that described in white wines. PMID- 12720413 TI - Disposition of doramectin milk residues in lactating dairy sheep. AB - Doramectin (DRM) is a broad spectrum macrocyclic lactone antiparasitic drug not approved for use in dairy animals. However, DRM and other endectocide compounds are widely used extra-label to control endo- and ectoparasites in dairy sheep. The plasma disposition kinetics and the pattern of DRM excretion in milk were characterized following its subcutaneous administration to lactating dairy sheep. DRM concentration profiles were measured in plasma and milk samples after validation of a specific HPLC-based methodology. DRM was detected between 1 h and 30 days post-treatment. DRM concentrations of 0.48 ng.mL(-1) (plasma) and 1.03 ng.mL(-1) (milk) were measured at 30 days post-treatment. DRM was extensively distributed from the bloodstream to the mammary gland, and large concentrations were excreted in milk. The peak concentrations and total amount of DRM recovered in milk (expressed as area under the concentration versus time curve) were 3-fold higher than those measured in plasma; 2.44% of the total DRM dose was excreted in milk. The long persistence of DRM milk residues should be seriously considered before its extra-label use in dairy animals is recommended. PMID- 12720414 TI - Polybrominated diphenyl ethers (PBDEs) in foodstuffs: human exposure through the diet. AB - Polybrominated diphenyl ethers (PBDEs) are used as flame retardants in a variety of materials, including synthetic polymers and textiles. Although these chemicals have been detected in environmental samples and human tissues, there is little information about human exposure to PBDEs through the diet. In the present study, we determined the concentrations of PBDEs in a number of food samples acquired in Catalonia (Spain) during 2000. The dietary intake of PBDEs was estimated for the general population living in this Spanish region. The highest PBDE concentrations were found in oils and fats, fish and shellfish, meat and meat products, and eggs, while the lowest levels corresponded to fruits, vegetables, and tubers. The dietary intake of PBDEs for an adult male was 97.3 ng/day (assuming not detected (ND) = (1)/(2) limit of detection (LOD)) or 81.9 ng/day (assuming ND = 0) The greatest contribution to these values corresponded to fish and shellfish, with approximately one-third of the total intake. TetraBDEs and pentaBDEs were the homologues showing the highest percentages of contribution to the sum of total PBDEs. The comparison of the current dietary intake with the suggested lowest observed adverse effect level value of 1 mg/kg/day for the most sensitive endpoints for toxic effects of PBDEs results in a safety factor over 5 orders of magnitude in relation to PBDE exposure from food. PMID- 12720416 TI - Alternating DNA and pi-conjugated sequences. Thermophilic foldable polymers. AB - Foldable polymers with alternating single-strand deoxyribonucleic acid and planar conjugated organic perylene tetracarboxylic diimide units were found to self organize into loosely folded nanostructures. Upon heating, the loosely folded structures become more ordered as evidenced by pi-stacking in the perylene segments. The folding and unfolding processes driven by the molecular interactions of adjacent perylenes were monitored in both aqueous and organic solutions. Heat-promoted folding, or inverse temperature behavior, which originates from positive enthalpy changes, was only observed in water. Therefore, we attributed this inverse temperature dependence to hydrophobic effects rather than pi-pi molecular orbital overlap between the perylene planes. These findings shed light on the design of new thermophiles in protein engineering as well as the construction of macromolecular-based nanodevices with actuator and sensory properties. PMID- 12720417 TI - Efficient epoxidation of electron-deficient olefins with a cationic manganese complex. AB - The complex [MnII(R,R-mcp)(CF3SO3)2] is an efficient and practical catalyst for the epoxidation of electron-deficient olefins. This catalyst is capable of epoxidizing olefins with as little as 0.1 mol % catalyst in under 5 min using 1.2 equiv of peracetic acid as the terminal oxidant. A wide scope of substrates are epoxidized including terminal, tertiary, cis and trans internal, enones, and methacrylates with >85% isolated yields. PMID- 12720419 TI - Fabrication of silica nanotube arrays from vertical silicon nanowire templates. AB - A simple thermal oxidation-etching process was developed to translate vertical silicon nanowire arrays into silica nanotube arrays. The obtained nanotubes perfectly retain the orientation of original silicon nanowire arrays. The inner tube diameter ranges from 10 to 200 nm. High-temperature oxidation produces relative thick, rigid, and pinhole-free walls that are made of condensed silica. This method could be useful for fabrication of single nanotube sensors and nanofluidic systems. PMID- 12720418 TI - Quantification of protein-ligand interactions by mass spectrometry, titration, and H/D exchange: PLIMSTEX. AB - Protein-ligand binding and the concomitant conformational change in the protein are of crucial importance in biophysics and drug design. We report a novel method to quantify protein-ligand interactions in solution by mass spectrometry, titration, and H/D exchange (PLIMSTEX). The approach can determine the conformational change, binding stoichiometry, and affinity in protein-ligand interactions including those that involve small molecules, metal ions, and peptides. Binding constants obtained by PLIMSTEX for four model protein-ligand systems agree with K values measured by conventional methods. At higher protein concentration, the method can be used to determine quickly the binding stoichiometry and possibly the purity of proteins. Taking advantage of concentrating the protein on-column and desalting, we are able to use different concentrations of proteins, buffer systems, salts, and pH in the exchange protocol. High picomole quantities of proteins are sufficient, offering significantly better sensitivity than that of NMR and X-ray crystallography. Automation could make PLIMSTEX a high throughput method for library screening, drug discovery, and proteomics. PMID- 12720420 TI - Metallocene polymerization catalyst ion-pair aggregation by cryoscopy and pulsed field gradient spin-echo NMR diffusion measurements. AB - Pulsed field gradient spin-echo (PGSE) NMR and cryoscopic measurements have been performed on a series of homogeneous metallocene polymerization catalyst ion pairs to determine if aggregation is a significant phenomenon under typical polymerization conditions. Cryoscopic measurements on [(Me5Cp)2ZrMe]+[MeB(C6F5)3] (1), [rac-Et(Indenyl)2ZrMe]+[MeB(C6F5)3]- (2), [(1,2 Me2Cp)2ZrCHTMS2]+[MeB(C6F5)3]- (3), [Me2Si(Me4Cp)(t-BuN)TiMe]+[MeB(C6F5)3]- (4), [Me2Si(Me4Cp)(t-BuN)ZrMe]+[MeB(C6F5)3]- (5), and [Me2C(Fluorenyl)(Cp)ZrMe]+[MeB(C6F5)3]- (6) were carried out in benzene in the 10 18 millimolal concentration range. PGSE measurements, using (p-tolyl)4Si as an internal standard, were also performed on catalyst ion-pairs 1, 4, 6, [(Me5Cp)2ThMe]+[B(C6F5)4]- (7), [(Me2SiCp2)ZrMe]+[MeB(C6F5)3]- (8), and [Cp2ZrMe]+[MeB(C6F5)3]- (9) in the 0.8-10.0 millimolar range. All results are consistent with a 1:1 ion-pair structural model and show little evidence for ion quadruples or higher-order aggregates. PMID- 12720421 TI - Excitation energy transfer in branched dendritic macromolecules at low (4 k) temperatures. AB - To understand the mode of energy transport in branched dendritic macromolecules, the optical excitation of a dendritic core (A-DSB) at low temperature (4.2 K) was investigated. Fluorescence depolarization measurements were utilized to probe the energy-transfer processes in the branching center at several different temperatures. We found that the anisotropy decay shows an interesting trend at low temperature where depolarization times decreased and the residual anisotropy value also decreased with decreasing temperature. The very fast anisotropy decay suggests a coherent mechanism of energy transport in these systems at low temperature. The contribution of inhomogeneous broadening is suggested as an important factor in the temperature dependence of the anisotropy decay and residual value. The change in inhomogeneous linewidth is responsible for this type of anisotropy behavior. PMID- 12720422 TI - Carboxylate-substituted radicals from phenylselenide derivatives. Designs on models for coenzyme B12-dependent enzyme-catalyzed rearrangements. AB - Laser flash photolysis (266 nm) of alpha- and beta-phenylselenyl esters, carboxylic acids, and carboxylates in aqueous acetonitrile media gave the corresponding radicals by homolytic cleavage of the phenylselenyl groups. In the beta-substituted systems, acid and carboxylate radicals reacted in intramolecular reporter reactions with approximately equal rate constants. For the alpha substituted systems, an ester- and carboxylic acid-substituted radical reacted in an intramolecular reporter reaction with the same rate constants, but the analogous alpha-carboxylate radical, a radical anion, reacted an order of magnitude less rapidly and with an activation energy that is 3 kcal/mol greater than that found for analogues. A kinetic titration of the equilibrating alpha acid and alpha-carboxylate radicals gave pKa = 4.6. The results indicate that alpha-ester and alpha-carboxylic acid radicals are unlikely to be appropriate models for alpha-carboxylate radicals, the intermediates formed in a large subset of coenzyme B12-dependent enzyme-catalyzed reactions. PMID- 12720423 TI - Novel small organic molecules for a highly enantioselective direct aldol reaction. AB - Novel organic molecules containing an l-proline amide moiety and a terminal hydroxyl for catalyzing direct asymmetric aldol reactions of aldehydes in neat acetone are designed and prepared. Catalyst 3d, prepared from l-proline and (1S,2S)-diphenyl-2-aminoethanol, exhibits high enantioselectivities of up to 93% ee for aromatic aldehydes and up to >99% ee for aliphatic aldehydes. A theoretical study of transition structures demonstrates the important role of the terminal hydroxyl group in the catalyst in the stereodiscrimination. Our results suggest a new strategy in the design of new organic catalysts for direct asymmetric aldol reactions and related transformations because plentiful chiral resources containing multi-hydrogen bond donors, for example, peptides, might be adopted in the design. PMID- 12720424 TI - Determination of an acidic scale in room temperature ionic liquids. AB - The acidity scale of different Bronsted acids in ionic liquids such as [BMIM][NTf2], [BMIM][BF4], and [BMMIM][BF4] has been investigated by determination of Hammett functions, using a spectrophotometric indicator method. This scale should permit one to correlate the acidity strength of ionic liquid systems with their ability to achieve acid-catalyzed reactions. PMID- 12720425 TI - Evidence of surfactant-induced formation of transient pores in lipid bilayers by using magnetic-fluid-loaded liposomes. AB - It is often assumed that surfactant-induced permeability of lipid membranes obeys a pore-formation mechanism, but, to date, this has not been totally proven. A novel approach is developed using a magnetic fluid composed of calibrated nanocrystals of maghemite (gamma-Fe2O3) as a permeability marker. It is shown that low amounts of surfactant molecules catalyze the transient opening of unilamellar phospholipid vesicles which permit the passage of 8 nm maghemite nanospheres before closing up. PMID- 12720426 TI - Protein assembly by orthogonal chemical ligation methods. AB - Chemical synthesis harbors the potential to provide ready access to natural proteins as well as to create nonnatural ones. The Staudinger ligation of a peptide containing a C-terminal phosphinothioester with a peptide containing an N terminal azide gives an amide with no residual atoms. This method for amide bond formation is orthogonal and complementary to other ligation methods. Herein, we describe the first use of the Staudinger ligation to couple peptides on a solid support. The fragment thus produced is used to assemble functional ribonuclease A via native chemical ligation. The synthesis of a protein by this route expands the versatility of chemical approaches to protein production. PMID- 12720427 TI - Observation of symmetry lowering and electron localization in the doublet-states of a spin-frustrated equilateral triangular lattice: Cu3(O2C16H23) x 1.2C6H12. AB - Cu3(O2C16H23)6.1.2C6H12, containing a Cu36+ core in an equilateral triangle geometry, has been found to be a versatile model system for investigating the spin-frustration phenomenon in a triangular lattice. It affords well-resolved EPR spectra from both of the two possible (Stotal = 1/2 and 3/2) spin states of the Cu36+ core. From 295 to 100 K, the spectra consist of a triplet, but with the central line overlapped by an additional, sharp peak, which replaces the triplet at 30 K and below. The triplet was thus assigned to the excited state with Stotal = 3/2, located at 324 +/- 5 K ( approximately 225 cm-1), with the zero-field parameters D = -535 G, E = 0, g parallel = 2.209 and g perpendicular = 2.057. The singlet was attributed to the Stotal = 1/2 state, with gxx = 2.005, gyy = 2.050, gzz = 2.282, and, surprisingly, a hyperfine splitting arising from a single Cu2+ nucleus, with Azz = 157 G. The detailed magnetic measurements on a three electron, equilateral triangular system, and the observation of symmetry lowering in the doublet ground state, should be of broad theoretical and experimental interest in molecular magnetism. PMID- 12720428 TI - First Cr(III)-SNS complexes and their use as highly efficient catalysts for the trimerization of ethylene to 1-hexene. AB - Cr(III) complexes of tridentate SNS ligands have been prepared and evaluated as catalysts for ethylene trimerization, with several giving very high activity and excellent selectivity toward 1-hexene when activated with methylaluminoxane. The new complexes illustrate the potential of sulfur-based ligands on early transition metals for catalysis. PMID- 12720429 TI - Selective C-arylation of free (NH)-heteroarenes via catalytic C-H bond functionalization. AB - A new system for palladium-catalyzed arylation of a broad spectrum of free (NH) heteroarenes has been developed (indole, pyrrole, pyrazole, 2-phenylimidazole, imidazole, benzimidazole, and purine). Remarkable selectivity has been achieved in the presence of MgO base, providing single C-arylation products, while no N arylation and no bis-arylation products have been detected. In the case of free imidazole, exclusive C-4 arylation may be switched to exclusive 2-arylation by the addition of CuI to the Pd/Ph3P/MgO system. When free aryl-(NH)-azoles are desired, direct arylation eliminates three steps in comparison to standard methods, including N-protection, stoichiometric metalation or halogenation, and N deprotection. PMID- 12720430 TI - Size-selective organization of enthalpic compatibilized nanocrystals in ternary block copolymer/particle mixtures. AB - Dependent on the relative particle core size, two distinct types of particle topologies in block copolymer/nanocrystal blends have been identified, that is, the localization of particles along the intermaterial dividing surface or at the center of the respective polymer domain. In ternary systems consisting of block copolymer and two different-sized nanocrystal species, the distinct morphological types are conserved, resulting in autonomous size-selective separation and organization of the respective nanocrystals within alternating arrays and sheets. PMID- 12720431 TI - Subpicosecond photoinduced charge injection from "molecular tripods" into mesoporous TiO2 over the distance of 24 angstroms. AB - Extended rigid tripodal sensitizers were used to investigate the rate of long distance photoinduced charge transfer from the MLCT excited states of RuII-based chromophores into mesoporous TiO2 films. The distance between the RuII center and the surface of the semiconductor was 24 A. Rapid biexponential charge injection with a major subpicosecond component as fast as 240 fs was observed upon femtosecond laser excitation of the tripods bound to the TiO2 surface. This rate exceeds the typical rates of vibrational cooling and thus strongly supports the possibility of "hot electron injection" occurring at very large donor-to semiconductor distances. PMID- 12720432 TI - A water-soluble and "self-assembled" polyoxometalate as a recyclable catalyst for oxidation of alcohols in water with hydrogen peroxide. AB - We have demonstrated that a simply prepared water-soluble polyoxometalate, Na12[WZnZn2(H2O)2(ZnW9O34)2], synthesized from readily available zinc and tungsten salts in the presence of nitric acid, is an effective catalyst for selective alcohol oxidation with hydrogen peroxide in biphasic (water-alcohol) reaction media. Experiments have shown that the "self-assembled" catalyst in its mother liquor was as active as the isolated catalyst. The aqueous catalyst solution is easily separated from the water-insoluble products and can be recycled without loss in activity or selectivity. PMID- 12720433 TI - 4H-1,2-benzoxazines with electron-withdrawing substituents on the benzene ring: synthesis and application as potent intermediates for oxygen-functionalized aromatic compounds. AB - A new general method for synthesizing functionalized 4H-1,2-benzoxazine derivatives is described. Although 4H-1,2-benzoxazine is one of the fundamental structure of the oxazine group, no general synthetic method for the heterocycle has been established. We found that 3-methoxycarbonyl-4H-1,2-benzoxazine was obtained in good yield when methyl 2-nitro-3-phenylpropionate was treated with an excess amount of trifluoromethanesulfonic acid. This methodology was also applicable for the synthesis of 4H-1,2-benzoxazine rings functionalized with various electron-withdrawing substituents on the benzene ring. Furthermore, we also show that the resulting 4H-1,2-benzoxazines can be used as precursors of functionalized o-quinone methides and multisubstituted phenols. This type of heterocycle can be a potent intermediate to oxygen-fuctionalized aromatic compounds. PMID- 12720434 TI - Titanium disulfide nanotubes as hydrogen-storage materials. AB - TiS2 nanotubes, which were synthesized through a chemical transport reaction, are very effective in reversible hydrogen absorption and desorption with the capacity of 2.5 wt %. PMID- 12720435 TI - Synthesis and spectroscopic properties of a series of beta-blocked long oligothiophenes up to the 96-mer: revaluation of effective conjugation length. AB - A series of extraordinarily long oligothiophenes up to the 96-mer has been developed by iterative oxidative coupling of the completely beta-blocked sexithiophene. They are highly conjugated like nonsubstituted oligothiophenes, and the effective conjugation of this system is extended to 96 thiophene units and much longer than that previously speculated for polythiophenes. PMID- 12720436 TI - Polymer catalysts from polymerization catalysts: direct encapsulation of metal catalyst into star polymer core during metal-catalyzed living radical polymerization. AB - Star polymers containing ruthenium complex in the core were prepared by ruthenium catalyzed living radical polymerization, where the metal catalysts were directly encapsulated on linking reactions of living poly(MMA) in the presence of ethylene glycol dimethacrylate as a linker and diphenyl-4-styrylphosphine as a ligand incorporated in the core. The products were characterized by SEC/MALLS, UV-vis, NMR, AFM, TEM, and ICP-AES and were employed as polymer catalysts for the oxidation reaction of alcohol. PMID- 12720437 TI - Chemical lithography of a conductive polymer using a traceless removable group. AB - Polyaniline could be easily converted into nitrosated polyaniline by reaction with nitrite ion in acids. The product is soluble in common solvents and could be deposited into thin films. The nitrosated polyaniline could be back-converted into polyaniline by acid hydrolysis. On the basis of those properties, a simple chemical lithographic process to produce conductive polyaniline images is demonstrated. PMID- 12720438 TI - Microlens formation in microgel/gold colloid composite materials via photothermal patterning. AB - We report on the nature of photothermally patterned regions inside self-assembled hydrogel nanoparticle materials containing coassembled colloidal Au. These composite materials are prepared from approximately 226-nm diameter particles composed of the environmentally responsive polymer, poly(N-isopropylacrylamide) (pNIPAm). Upon centrifugation to achieve a proper volume fraction, these close packed assemblies display a sharp Bragg diffraction peak in the midvisible region of the spectrum and can be reversibly converted into a nondiffracting glassy material as the temperature is raised above the characteristic phase transition temperature of the polymer. The addition of 16-nm colloidal Au prior to centrifugation allows the homogeneous distribution of metal nanoparticles throughout the close-packed material. Localized heating is then possible upon excitation of the Au plasmon absorption with a frequency doubled Nd:YAG laser (lambda = 532 nm). Such localized heating events lead to patterned regions of ordered crystalline phases inside of bulk glassy phases. We illustrate that the nature of the locally patterned area results in the formation of a microlens due to density/refractive index gradient in the patterned crystalline region. The Gaussian power distribution of the incident beam is thought to be a contributing factor in the microlens formation. Microlens formation is shown by observing interference patterns similar to Newton's rings, which change over time as the region is formed. A true hallmark of the lens is also demonstrated by focusing an image through the patterned structure. PMID- 12720439 TI - A miniaturized, parallel, serially diluted immunoassay for analyzing multiple antigens. AB - This paper describes a microfluidic immunoassay that is applicable to the parallel determination of multiple analytes and that requires only a few microliters of sample. This assay relies on a microchannel network that achieves serial dilution of analytes; this network replaces manual dilutions employed in traditional immunoassays and enables the analysis of multiple analytes simultaneously. The immunoassay was demonstrated by an analysis of concentrations of antibodies against the HIV viral proteins gp120 and gp41 in human serum. PMID- 12720440 TI - Spirastrellolide A, an antimitotic macrolide isolated from the Caribbean marine sponge Spirastrella coccinea. AB - Sprirastrellolide A, a novel antimitotic macrolide, has been isolated from the Caribbean marine sponge Spirastrella coccinea. It has a 47-carbon linear polyketide backbone incorporated into a highly functionalized 38-membered lactone containing a tetrahydropyran and two spiro bispyran substructures embedded in the macrocycle and a side chain terminating in a carboxylic acid. Sprirastrellolide A's structure was elucidated by detailed spectroscopic analysis of its methyl ester 2. Spirastrellolide A methyl ester 2 shows potent activity in a cell-based assay that detects mitotic arrest, but it does not affect tubulin polymerization in vitro. It has the unusual biological property of being able to accelerate the entry of cells into mitosis from other cell-cycle stages, before it arrests them in mitosis. PMID- 12720441 TI - An unnatural hydrophobic base pair with shape complementarity between pyrrole-2 carbaldehyde and 9-methylimidazo[(4,5)-b]pyridine. AB - An unnatural hydrophobic base, pyrrole-2-carbaldehyde (denoted as Pa), was developed as a specific pairing partner of 9-methylimidazo[(4,5)-b]pyridine (Q). The Q base is known to pair with 2,4-difluorotoluene (F) as an isostere of the A T pair, and F also pairs with A efficiently in replication. In contrast, the Q-Pa pair showed specific selectivity in replication, and the five-membered-ring base Pa paired efficiently with Q but paired poorly with A. In addition, the interaction of Pa with DNA polymerases was superior, in comparison to that of F. The aldehyde group of Pa was recognized well by the Klenow fragment of Escherichia coli DNA polymerase I and the reverse transcriptase of Avian myeloblastosis virus. The structural features of the Q-Pa pair in a DNA duplex were analyzed by NMR, showing the shape complementarity of the Pa fitting with Q. The structurally unique base Pa provides valuable information for the development of unnatural base pairs toward the expansion of the genetic alphabet. PMID- 12720442 TI - Aromatic-aromatic interactions in crystal structures of helical peptide scaffolds containing projecting phenylalanine residues. AB - Aromatic-aromatic interactions between phenylalanine side chains in peptides have been probed by the structure determination in crystals of three peptides: Boc-Val Ala-Phe-Aib-Val-Ala-Phe-Aib-OMe, I; Boc-Val-Ala-Phe-Aib-Val-Ala-Phe-Aib-Val-Ala Phe-Aib-OMe, II; Boc-Aib-Ala-Phe-Aib-Phe-Ala-Val-Aib-OMe, III. X-ray diffraction studies reveal that all three peptides adopt helical conformations in the solid state with the Phe side chains projecting outward. Interhelix association in the crystals is promoted by Phe-Phe interactions. A total of 15 unique aromatic pairs have been characterized in the three independent crystal structures. In peptides I and II, the aromatic side chains lie on the same face of the helix at i/i + 4 positions resulting in both intrahelix and interhelix aromatic interactions. In peptide III, the Phe side chains are placed on the opposite faces of the helix, resulting in exclusive intermolecular aromatic interactions. The distances between the centroids of aromatic pair ranges from 5.11 to 6.86 A, while the distance of closest approach of ring carbon atoms ranges from 3.27 to 4.59 A. Examples of T-shaped and parallel-displaced arrangements of aromatic pairs are observed, in addition to several examples of inclined arrangements. The results support the view that the interaction potential for a pair of aromatic rings is relatively broad and rugged with several minima of similar energies, separated by small activation barriers. PMID- 12720443 TI - Parallel synthesis and biophysical characterization of a degradable polymer library for gene delivery. AB - We recently reported the parallel synthesis of 140 degradable poly(beta-amino esters) via the conjugate addition of 20 primary or secondary amine monomers to seven different diacrylate monomers. To explore possible structure/function relationships and further characterize this class of materials, we investigated the ability of each DNA-complexing polymer to overcome important cellular barriers to gene transfer. The majority of vectors were found to be uptake limited, but complexes formed from polymers B14 and G5 displayed high levels of internalization relative to "naked" DNA (18x and 32x, respectively). Effective diameter and zeta potential measurements indicated that, in general, small particle size and positive surface charge led to higher internalization rates. Of the 10 DNA/polymer complexes with the highest uptake levels, all had effective diameters less than 250 nm and nine had positive zeta potentials. Lysosomal trafficking was investigated by measuring the pH environment of delivered DNA. Complexes prepared with polymers G5, G10, A13, B13, A14, and B14 were found to have near neutral pH measurements, suggesting that they were able to successfully avoid trafficking to acidic lysosomes. This work highlights the value of parallel synthesis and screening approaches for the discovery of new polymers for gene delivery and the elucidation of structure/function relationships for this important class of materials. PMID- 12720444 TI - Measurement of submicrosecond intramolecular contact formation in peptides at the single-molecule level. AB - We describe a single-molecule-sensitive method to determine the rate of contact formation and dissociation between tryptophan and an oxazine derivative (MR121) on the basis of measurements of the photon distance distribution. Two short peptides (15 and 20 amino acids) derived from the transactivation domain of the human oncoprotein p53 were investigated. With the fluorophore attached at the N terminal end of the flexible peptides, fluorescence of the dye is efficiently quenched upon contact formation with a tryptophan residue. The mechanism responsible for the efficient fluorescence quenching observed in the complexes is assumed to be a photoinduced electron-transfer reaction occurring predominantly at van der Waals contact. Fluorescence fluctuations caused by intramolecular contact formation and dissociation were recorded using confocal fluorescence microscopy with two avalanche photodiodes and the time-correlated single-photon counting technique, enabling a temporal resolution of 1.2 ns. Peptides containing a tryptophan residue at positions 9 and 8, respectively, show contact formation with rate constants of 1/120 and 1/152 ns(-1), respectively. Whereas the rate constants of contact formation most likely directly report on biopolymer chain mobility, the dissociation rate constants of 1/267 and 1/742 ns(-1), respectively, are significantly smaller and reflect strong hydrophobic interactions between the dye and tryptophan. Fluorescence experiments on point mutated peptides where tryptophan is exchanged by phenylalanine show no fluorescence quenching. PMID- 12720445 TI - Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometric analysis of metal-ion selected dynamic protein libraries. AB - The application of electrospray ionization (ESI) Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry to the investigation of the relative stabilities (and thus packing efficiencies) of Fe-bound trihelix peptide bundles is demonstrated. Small dynamic protein libraries are created by metal-ion assisted assembly of peptide subunits. Control of the trimeric aggregation state is coupled to stability selection by exploiting the coordination requirements of Fe(2+) in the presence of bidentate 2,2'-bipyridyl ligands covalently appended to the peptide monomers. At limiting metal-ion concentration, the most thermodynamically stable, optimally packed peptide trimers dominate the mass spectrum. The identities of optimally stable candidate trimers observed in the ESI FT-ICR mass spectra are confirmed by resynthesis of exchange-inert analogues and measurement of their folding free energies. The peptide composition of the trimers may be determined by infrared multiphoton dissociation (IRMPD) MS(3) experiments. Additional sequence information for the peptide subunits is obtained from electron capture dissociation (ECD) of peptides and metal-bound trimers. The experiments also suggest the presence of secondary structure in the gas phase, possibly due to partial retention of the solution-phase coiled coil structure. PMID- 12720446 TI - Strings of vesicles: flow behavior in an unusual type of aqueous gel. AB - This is a study of 10 asymmetric gemini surfactants that self-assemble into vesicles which, in turn, self-assemble into gels. The geminis have the following general structure: long-chain/phosphate/2-carbon spacer/quaternary nitrogen/short chain. Dynamic light scattering and transmission electron microscopy (TEM) demonstrate that in dilute aqueous systems these compounds self-assemble into vesicles. The vesicles are cohesive as proven by cryo-high resolution electron microscopy (cryo-HRSEM) images that reveal a "pearls on a string" morphology. These strings of vesicles create a complex network that rigidifies the water. The one gemini in the study that does not form a gel is also the only vesicle system that, according to cryo-HRSEM and TEM, assembles into clumps rather than chains. It is proposed that the vesicles are cohesive owing to protrusion of short chains from the vesicle surfaces, thereby creating hydrophobic "patches" whose intervesicular overlap supersedes the normal membrane/membrane repulsive forces. Analogous geminis having two long chains, neither of which are thought capable of departing from their bilayers, also form vesicles, but they are noncohesive (as expected from the model). Rheological experiments carried out on the gels show that gelation is mechanically reversible. Thus, if an applied torque breaks a string, the string can rapidly mend itself as long as the temperature exceeds its calorimetrically determined T(m) value. Gel strength, as manifested by the yield stress of the soft material, was shown to be particularly sensitive to the structure of the gemini. All three individual components of the systems (geminis, vesicles, and gels) have widespread practical applications. PMID- 12720447 TI - In vitro evolution of an RNA-cleaving DNA enzyme into an RNA ligase switches the selectivity from 3'-5' to 2'-5'. AB - Deoxyribozymes that ligate RNA expand the scope of nucleic acid catalysis and allow preparation of site-specifically modified RNAs. Previously, deoxyribozymes that join a 5'-hydroxyl and a 2',3'-cyclic phosphate were identified by in vitro selection from random DNA pools. Here, the alternative strategy of in vitro evolution was used to transform the 8-17 deoxyribozyme that cleaves RNA into a family of DNA enzymes that ligate RNA. The parent 8-17 DNA enzyme cleaves native 3'-5' phosphodiester linkages but not 2'-5' bonds. Surprisingly, the new deoxyribozymes evolved from 8-17 create only 2'-5' linkages. Thus, reversing the direction of the DNA-mediated process from ligation to cleavage also switches the selectivity in forming the new phosphodiester bond. The same change in selectivity was observed upon evolution of the 10-23 RNA-cleaving deoxyribozyme into an RNA ligase. The DNA enzymes previously isolated from random pools also create 2'-5' linkages. Therefore, deoxyribozyme-mediated formation of a non native 2'-5' phosphodiester linkage from a 5'-hydroxyl and a 2',3'-cyclic phosphate is strongly favored in many different contexts. PMID- 12720448 TI - Synthesis and characterization of cross-linked reverse micelles. AB - The design and synthesis of a new cross-linkable amphiphile is reported. Solutions of the amphiphile in a toluene/water mixture form reverse micelles as indicated by dynamic light scattering and NMR spectroscopy. As indicated by dynamic light scattering, TEM, and NMR spectroscopy data, these reverse micelles can be cross-linked without drastically changing the radius of the reverse micelles. Mixed reverse micelles are also characterized and cross-linked. The cross-linked reverse micelles are demonstrated to facilitate phase transfer and can be used to site isolate a catalyst. PMID- 12720449 TI - Ruthenium(II) dendrimers containing carbazole-based chromophores as branches. AB - Three new luminescent and redox-active Ru(II) complexes containing novel dendritic polypyridine ligands have been synthesized, and their absorption spectra, luminescence properties (both at room temperature in fluid solution and at 77 K in rigid matrix), and redox behavior have been investigated. The dendritic ligands are made of 1,10-phenanthroline coordinating subunits and of carbazole groups as branching sites. The first and second generation species of this novel class of dendritic ligands (L1 and L2, respectively; see Figure 1 for their structural formulas) have been prepared and employed. The metal dendrimers investigated are [Ru(bpy)(2)(L1)](2+) (1; bpy = 2,2'-bipyridine), [Ru(bpy)(2)(L2)](2+) (2), and [Ru(L1)(3)](2+) (3; see Figure 2). For the sake of completeness and comparison purposes, also the absorption spectra, redox behavior, and luminescence properties of L1 and L2 have been studied, together with the properties of 3,6-di(tert-butyl)carbazole (L0) and [Ru(bpy)(2)(phen)](2+) (4, phen = 1,10-phenanthroline). The absorption spectra of the free dendritic ligands show features which can be assigned to the various subunits (i.e., carbazole and phenanthroline groups) and additional bands at lower energies (at lambda > 300 nm) which are assigned to carbazole-to phenanthroline charge-transfer (CT) transitions. These latter bands are significantly red-shifted upon acid and/or zinc acetate addition. Both L1 and L2 exhibit relatively intense luminescence at room temperature in fluid solution (lifetimes in the nanosecond time scale, quantum yields of the order of 10(-2) 10(-1)) and at 77 K in rigid matrix (lifetimes in the millisecond time scale). Such a luminescence is assigned to CT states at room temperature and to phenanthroline-centered pi-pi triplet levels at 77 K. The room-temperature luminescence of L1 and L2 is totally quenched by acid or zinc acetate. The metal dendrimers exhibit the typical absorption and luminescence properties of Ru(II) polypyridine complexes. In particular, metal-to-ligand charge-transfer (MLCT) bands dominate the visible absorption spectra, and formally triplet MLCT levels govern the excited-state properties. Excitation spectroscopy evidences that all the light absorbed by the dendritic branches is transferred with unitary efficiency to the luminescent MLCT states in 1-3, showing that the new metal dendrimers can be regarded as efficient light-harvesting antenna systems. All the free ligands and metal dendrimers exhibit a rich redox behavior (except L2 and 3, whose redox behavior was not investigated because of solubility reasons), with clearly attributable reversible carbazole- and metal-centered oxidation and polypyridine-centered reduction processes. The electronic interaction between the carbazole redox-active sites of the dendritic ligands is affected by Ru(II) coordination. PMID- 12720451 TI - The azulene-to-naphthalene rearrangement revisited: a DFT study of intramolecular and radical-promoted mechanisms. AB - Intramolecular and radical-promoted mechanisms for the rearrangement of azulene to naphthalene are assessed with the aid of density functional calculations. All intramolecular mechanisms have very high activation energies (>/=350 kJ mol(-1) from azulene) and so can only be competitive at temperatures above 1000 degrees C. Two radical-promoted mechanisms, the methylene walk and spiran pathways, dominate the reaction below this temperature. The activation energy for an orbital symmetry-allowed mechanism via a bicyclobutane intermediate is 382 kJ mol(-1). The norcaradiene-vinylidene mechanism that has been proposed in order to explain the formation of small amounts of 1-phenyl-1-buten-3-ynes from flash thermolysis of azulene has an activation energy of 360 kJ mol(-1); subtle features of the B3LYP/6-31G(d) energy surface for this mechanism are discussed. All intermediates and transition states on the spiran and methylene walk radical promoted pathways have been located at the B3LYP/6-31G(d) level. Interconversion of all n-H-azulyl radicals via hydrogen shifts was also examined, and hydrogen shifts around the five-membered ring are competitive with the mechanisms leading to rearrangement to naphthalene, but those around the seven-membered ring are not. Conversion of a tricyclic radical to the 9-H-naphthyl radical is the rate limiting transition state on the spiran pathway, and lies 164.0 kJ mol(-1) above that of the 1-H-azulyl radical. The transition state for the degenerate hydrogen shift between the 9-H-azulyl and 10-H-azulyl radicals is 7.4 kJ mol(-1) lower. Partial equilibration of the intermediates in the spiran pathway via this shift may therefore occur, and this can account for the surprising formation of 1 methylnaphthalene from 2-methylazulene. The rate-limiting transition state for the methylene walk pathway involves the concerted transfer of a methylene group from one ring to the other and lies 182.3 kJ mol(-1) above that of the 1-H-azulyl radical. It is shown that rearrangement via a combination of 31% methylene walk and 69% spiran pathways can account semiquantitatively for all the products from 1-(13)C-azulene, 9-(13)C-azulene, and 4,7-(13)C(2)-azulene, in addition to accounting for the products from methylazulenes, and the formation of naphthalene d(0) and -d(2) from azulene-4-d. It is also pointed out that a small extension to the spiran pathway could provide an alternative explanation for the formation of 1-phenyl-1-buten-3-ynes. PMID- 12720450 TI - Mechanistic analysis of acyl transferase domain exchange in polyketide synthase modules. AB - Many polyketides are synthesized by a class of multifunctional enzymes called type I modular polyketide synthases (PKSs). Several reports have described the power of predictively altering polyketide structure by replacing individual PKS domains with homologues from other PKSs. For example, numerous erythromycin analogues have been generated by replacing individual methylmalonyl-specific acyl transferase (AT) domains of the 6-deoxyerythronolide B synthase (DEBS) with malonyl-, ethylmalonyl-, or methoxymalonyl-specific domains. However, the construction of hybrid PKS modules often attenuates product formation both kinetically and distributively. The molecular basis for this mechanistic imperfection is not understood. We have systematically analyzed the impact of replacing an AT domain of DEBS on acyl-AT formation, acyl-CoA:HS-NAc acyl transferase activity, acyl-CoA:ACP acyl transferase activity (nucleophile charging), acyl-SNAc:ketosynthase acyl transferase activity (electrophile charging), and beta-ketoacyl ACP synthase activity (condensation). As usual, domain junctions were located in interdomain regions flanking the AT domain. Kinetic analysis of hybrid modules containing either malonyl transferase or methylmalonyl transferase domains revealed a 15-20-fold decrease in overall turnover numbers of the hybrid modules as compared to the wild-type module. In contrast, both the activity and the specificity of the heterologous AT domains remained unaffected. Moreover, no defects could be detected in the ability of the heterologous AT domains to catalyze acyl-CoA:ACP acyl transfer. Single turnover studies aimed at directly probing the ketosynthase-catalyzed reaction led to two crucial findings. First, wild-type modules catalyzed chain elongation with comparable efficiency regardless of whether methylmalonyl-ACP or malonyl-ACP were the nucleophilic substrates. Second, chain elongation in all hybrid modules tested was seriously attenuated relative to the wild-type module. Our data suggest that, as currently practiced, the most deleterious impact of AT domain swapping is not on the substrate specificity. Rather, it is due to the impaired ability of the KS and ACP domains in the hybrid module to catalyze chain elongation. Consistent with this proposal, limited proteolysis of wild-type and hybrid modules showed major differences in cleavage patterns, especially in the region between the KR and ACP domains. PMID- 12720452 TI - Concerning the denticity of the dimethylsulfinyl anion in Meisenheimer complexation. AB - The denticity (O-, S-, and C-nucleophilic reactivity) of the dimethylsulfinyl carbanion ("dimsyl") toward 1,3,5-trinitrobenzene (TNB) has been studied by NMR spectroscopy, and structures of adducts have been assigned. Three dimsyl adducts are observed for the first time and have been ascribed to the O-adduct 15, the S adduct 16, and the C-adduct 17. The kinetic (15 > 16 > 17) and thermodynamic preference (17 > 16 > 15) for the reactivity of dimsyl toward TNB is compared to the known O- and C-reactivity of enolate anions and the O- and S-reactivity of dimethyl sulfoxide. Thus, dimsyl apparently represents a unique system in which three adjacent atoms having unshared electron pairs can utilize these in covalent bond formation. PMID- 12720453 TI - An enantioselective synthesis of FR182877 provides a chemical rationalization of its structure and affords multigram quantities of its direct precursor. AB - The evolution of a strategy culminating in an efficient, enantioselective synthesis of the potent microtubule-stabilizing agent FR182877 is described. Guided by a proposed biogenesis of this complex natural product, a solution emerged that involved the first reported example of a double transannular Diels Alder reaction to fashion the key elements of its hexacyclic structure. This pivotal transformation creates a complex pentacycle from a 19-membered macrocyclic pentaene, forming seven new stereogenic centers in a fully diastereocontrolled fashion. The efficiency of the approach ultimately enabled the preparation of multigram quantities of the direct precursor of FR182877 for conversion to the relatively unstable natural product when required. The reactivity of the strained, bridgehead olefin of this secondary metabolite with biologically relevant nucleophiles is also described. PMID- 12720454 TI - Facile access to monodisperse ultralarge rings. AB - A facile access to monodisperse ultralarge rings counting 126, 174, and 294 ring atoms is described. It follows a reaction sequence that is well suited for the preparation of [2]catenanes but altered just in the sequence of the two steps cyclization and carbonate formation. The carbonate acts as a covalent template that is easily formed and later cleaved. The obtained monocyclic products are constitutional isomers of the catenanes. PMID- 12720455 TI - Total synthesis of (+)-asteltoxin. AB - A convergent synthesis of (+)-asteltoxin (1) has been achieved by the Horner Emmons olefination of bis(tetrahydrofuran) aldehyde 53 and alpha-pyrone phosphonate 5. A key step features the stereoselective construction of a sterically congested quaternary center embedded in the densely functionalized bis(tetrahydrofuran) subunit by a Lewis acid-catalyzed, pinacol-type rearrangement of an epoxy silyl ether. This pivotal rearrangement methodology parallels the proposed biosynthetic pathway of 1 and is ripe for applications to the stereocontrolled synthesis of structurally complex natural products. PMID- 12720456 TI - An experimental and theoretical study of the type C enone rearrangement: mechanistic and exploratory organic photochemistry. AB - We recently described a new photochemical rearrangement which we termed a Type C process. The reaction involves a delta to alpha aryl migration in 5-disubstituted cyclohexenones also having bulky C-3 substituents. In contrast to most cyclohexenone rearrangements, the reaction occurs via a twisted pi-pi excited triplet rather than the usual n-pi state. The electronic nature of the rearrangement was assessed using migration selectivity with p-anisyl and p cyanophenyl groups. A synthesis of the reactants was elaborated, and the product structures were established by X-ray and NMR analysis. The reaction mechanism was established further with DFT and CASSCF computations. In the latter, localized NBO basis orbitals permitted proper selection of the active space. The nature of the diradical intermediates as well as the transition states was established computationally. Sensitization experiments with regioselectivities the same as those in direct irradiation confirmed the triplet multiplicity of the process. PMID- 12720457 TI - Tyrosine 89 accelerates Co-carbon bond homolysis in methylmalonyl-CoA mutase. AB - The contribution of the active-site residue, Y89, to the trillion-fold acceleration of Co-carbon bond homolysis rate in the methylmalonyl-CoA mutase catalyzed reaction has been evaluated by site-directed mutagenesis. Conversion of Y89 to phenylalanine or alanine results in a 10(3)-fold diminution of k(cat) and suppression of the overall kinetic isotope effect. The spectrum of the enzyme under steady-state conditions reveals the presence of AdoCbl but no cob(II)alamin. Together, these results are consistent with homolysis becoming completely rate determining in the forward direction in the two mutants and points to the role of Y89 as a molecular wedge in accelerating Co-carbon bond cleavage. PMID- 12720459 TI - "Oscillating" metallocene catalysts: what stops the oscillation? AB - The 150 MHz (13)C NMR microstructural analysis of polypropylene samples produced with two representative "oscillating" metallocene catalysts of largely different steric hindrance, namely [(2-(3,5-di-tert-butyl-4 methoxyphenyl)indenyl)(2)ZrP](+) and [(2-phenylindenyl)(2)ZrP](+) (P = polymeryl), and the implications on the origin of the stereocontrol are presented and discussed in detail. The original mechanistic proposal of an "oscillation" between a rac-like (isotactic-selective) and a meso-like (nonstereoselective) conformation cannot explain the observed polymer configuration. The isotactic stereoblock nature of the polymers obtained with the former catalyst proves unambiguously that the active cation "oscillates" between the two enantiomorphous rac-like conformations at an average frequency that, even at high propene concentration, is only slightly lower than that of monomer insertion. The less hindered [(2-phenylindenyl)(2)ZrP](+) gives instead a largely stereoirregular polypropylene, which is the logical consequence of a faster ligand rotation; however, depending on the use conditions (in particular, on the nature of the cocatalyst and the polarity of the solvent), the polymerization products may also contain appreciable amounts of a fairly isotactic fraction. The peculiar microstructure of this fraction, with isotactic blocks of the same relative configuration spanned by very short atactic ones, rules out the possibility that the latter are due to an active species in meso-like conformation and points rather to a conformationally "locked" rac-like species with restricted ring mobility. The hypothesis of a stereorigidity induced by the proximity to a counteranion, which would play the role of the interannular bridge in the rac bis(indenyl) ansa-metallocenes, was tested by computer modeling on a [rac-(2 phenylindenyl)(2)ZrMe(C(3)H(6))][B(C(6)F(5))(4)] ion couple and found viable. PMID- 12720458 TI - Polyunsaturated dicarboxylate tethers connecting dimolybdenum redox and chromophoric centers: syntheses, structures, and electrochemistry. AB - Compounds with two quadruply bonded Mo(2)(4+) units, Mo(2)(DAniF)(3) (DAniF = N,N '-di-p-anisylformamidinate), linked by unsaturated dicarboxylate dianions of various lengths have been prepared and their spectroscopic and electrochemical properties studied. As identified by the dicarboxylate linkers, these compounds are maleate (7), allene-1,3-dicarboxylate (10), cis,cis-muconate (11), trans,trans-muconate (12), octa-2,4,6-trans,trans,trans-hexatriene-1,8-dioate (tamuate, 13), and deca-2,4,6,8-trans,trans,trans,trans-octatetraene-1,10-dioate (texate, 14). The latter three molecules complete the five-membered (all trans) series [Mo(2)(DAniF)(3)](O(2)C(CH=CH)(n)CO(2))[Mo(2)(DAniF)(3)] (n = 0-4). Several unsymmetrical paddlewheel compounds of the type Mo(2)(DAniF)(3)(O(2)CX) (X = C triple bond CH (3), CH=CH(2) (4), (E)-CH=CH-CH=CH(2) (5)) have also been prepared for comparison to the molecules in which there are linked Mo(2) units. The precursors [Mo(2)(DAniF)(3)(MeCN)(2)](BPh(4)), [1]BPh(4), and Mo(2)(DAniF)(3)Cl(MeCN) (2) have also been isolated and characterized. The structures of all new molecules have been established by X-ray crystallography, including the methyl esters of various carboxylates used as ligands. All of the linked molecules have been examined by cyclic and differential pulse voltammetry, and deltaE(1/2) values, the separation between successive Mo(2)(4+)/Mo(2)(5+) oxidations, have been determined. Those compounds with highly unsaturated, fully conjugated linkers demonstrate electrochemical communication from end-to-end that is more persistent over distance than is accounted for by an electrostatic interaction alone, implying that the pi system of these dicarboxylate linkers is mediating communication. In the series [Mo(2)(DAniF)(3)](O(2)C(CH=CH)(n)CO(2))[Mo(2)(DAniF)(3)] (n = 0-4), the first oxidation potential shifts progressively to less positive values due to an increasing contribution of the polyolefinic alpha,omega-dicarboxylate to the molecular orbital undergoing oxidation. This first oxidation potential approaches a limiting value of 63 mV (vs Ag/AgCl) as n becomes infinitely long. Compound 11 can be photoisomerized to 12 in a process that is affected by the presence of the Mo(2)(4+) units, as the analogous rearrangement of dimethyl cis,cis-muconate is faster. PMID- 12720460 TI - Non-localized ligand-to-metal charge transfer excited states in (Cp)2Ti(IV)(NCS)2. AB - The bent d(0) titanium metallocene (Cp)(2)Ti(NCS)(2) exhibits an intense phosphorescence from a ligand-to-metal charge transfer triplet excited state at 77 K in an organic glass substrate and a poly(methyl methacrylate) plastic substrate. Quantum chemical calculations and spectroscopic studies show that the orbital parentage of this triplet state arises from the promotion of an electron from an essentially nonbonding symmetry adapted pi molecular orbital located on the NCS(-) ligands to a d(z)2-(y)2 orbital located on the Ti metal. Standard infrared spectroscopy of (Cp)(2)Ti(NCS)(2) in its ground electronic state at 77 K reveals a pair of closely spaced absorptions at (2072 cm(-1), 2038 cm(-1))(glass) and (2055 cm(-1), 2015 cm(-1))(plastic) that are assigned, respectively, to the symmetric and antisymmetric CN stretching modes of the two coordinated NCS(-) ligands. Low-temperature (77 K) time-resolved infrared spectroscopy that accesses the phosphorescing triplet excited state on the ns time scale shows an IR bleach that is coincident with the two ground state CN stretching bands and an associated grow-in of a pair of new IR bands at slightly lower energies (2059 cm( 1), 2013 cm(-1))(glass) and (2049 cm(-1), 1996 cm(-1))(plastic) that are assigned, respectively, to the symmetric and antisymmetric CN stretches in the emitting triplet state. These transient IR bands decay with virtually identical lifetimes to those observed for the phosphorescence decays when measured under identical experimental conditions. Singular value decomposition analysis of the time-resolved infrared data shows that the observed transient IR features arise from the same electronic manifold as measured through luminescence studies. The close similarity between the ground state and excited-state CN stretching bands in (Cp)(2)Ti(NCS)(2) indicates that symmetry breaking does not occur in forming the charge-transfer triplet excited-state manifold; i.e., electron density is withdrawn from a delocalized pi MO spread across both NCS(-) ligands. Calculations at several levels of theory reveal a delocalized ligand-to-metal charge transfer excited triplet manifold. These calculations closely reproduce the relative intensity ratios and frequencies of the symmetric and antisymmetric transient infrared vibrations in the CN region. This study is the first time resolved infrared investigation of a ligand-to-metal charge-transfer excited state and the first to be performed at cryogenic temperatures in thin-film organic glass and plastic substrates. PMID- 12720461 TI - First soluble M@C60 derivatives provide enhanced access to metallofullerenes and permit in vivo evaluation of Gd@C60[C(COOH)2]10 as a MRI contrast agent. AB - M@C(60) and related endohedral metallofullerenes comprise a significant portion of the metallofullerene yield in the traditional arc synthesis, but their chemistry and potential applications have been largely overlooked because of their sparse solubility. In this work, procedures are described to solublize Gd@C(60) species for the first time by forming the derivative, Gd@C(60)[C(COOCH(2)CH(3))(2)](10), and its hydrolyzed water-soluble form, Gd@C(60)[C(COOH)(2)](10). Imparting water solubility to Gd@C(60) permits its evaluation as a magnetic resonance imaging (MRI) contrast agent. Relaxometry measurements for Gd@C(60)[C(COOH)(2)](10) reveal it to possess a relaxivity (4.6 mM(-1) s(-1) at 20 MHz and 40 degrees C) comparable to that of commercially available Gd(III) chelate-based MRI agents. An in vivo MRI biodistribution study in a rodent model reveals Gd@C(60)[C(COOH)(2)](10) to possess the first non reticuloendothelial system (RES) localizing behavior for a water-soluble endohedral metallofullerene species, consistent with its lack of intermolecular aggregation in solution as determined by light-scattering measurements. This first derivatization and use of a M@C(60) species suggests new potential for metallofullerene technologies by reducing reliance on the chromatographic purification procedures normally employed for the far less abundant M@C(82) and related endohedrals. The recognition that water-soluble fullerene derivatives can be designed to avoid high levels of RES uptake is an important step toward fullerene-based pharmaceutical development. PMID- 12720462 TI - Exfoliated nanosheets as a new strong solid acid catalyst. AB - Two-dimensional metal oxide sheets in HTiNbO(5) and HSr(2)Nb(3)O(10), cation exchangeable layered metal oxides, were examined as solid acid catalysts. Exfoliation of HTiNbO(5) and HSr(2)Nb(3)O(10) in aqueous solutions formed colloidal single-crystal TiNbO(5)(-) and Sr(2)Nb(3)O(10)(-) nanosheets, which precipitated under an acidic condition to form aggregates of HTiNbO(5) nanosheets and HSr(2)Nb(3)O(10) nanosheets. Although esterification of acetic acid, cracking of cumene, and dehydration of 2-propanol were not catalyzed by original HTiNbO(5) because of the narrow interlayer distance, which prevents the insertion of organic molecules, HTiNbO(5) nanosheets functioned as a strong solid acid catalyst for the reactions. Nanosheets of HSr(2)Nb(3)O(10) exhibited no or slight catalytic activity for these reactions. NH(3) temperature-programmed desorption and (1)H magic-angle spinning nuclear magnetic resonance spectroscopy revealed that HTiNbO(5) nanosheets have strong Bronsted acid sites, whereas HSr(2)Nb(3)O(10) nanosheets do not. PMID- 12720463 TI - Polyunsaturated dicarboxylate tethers connecting dimolybdenum redox and chromophoric centers: absorption spectra and electronic structures. AB - The absorption spectra of a series of compounds of the type [Mo(2)(DAniF)(3)](O(2)CXCO(2))[Mo(2)(DAniF)(3)] (DAniF = N,N '-di-p anisylformamidinate) have been measured and revealed a strong dependence of the electronic transitions and, therefore, the colors upon the chemical nature of the dicarboxylate linker. The more intense colors and lower energy absorptions are observed with those compounds having unsaturated dicarboxylate linkers. Static and time-dependent DFT calculations were undertaken to identify the electronic excitations responsible for the observed colors. For those compounds with chemically unsaturated and fully conjugated dicarboxylate linkers (oxalate, 6; fumarate, 8; acetylene dicarboxylate, 9; cis,cis-muconate, 11; trans,trans muconate, 12; tamuate, 13; texate, 14; terephthalate, 15), the lowest energy absorptions are Mo(2)(4+) delta --> dicarboxylate pi metal-to-ligand charge transfer transitions. Those compounds with chemically saturated linkers (succinate, 20) have delta --> delta transitions as their lowest energy absorptions with essentially independent and noninteracting Mo(2)(4+) chromophores. PMID- 12720464 TI - Vibrational spectroscopy of mechanically compressed monolayers. AB - Sum-frequency spectroscopy has been used to investigate the behavior of self assembled monolayers in a solid-solid contact. Various alkanethiol layers on gold were observed before, during, and after compression to 660 MPa against a sapphire counterface. Well-ordered layers that differ only in the length of their alkane tails (C(8) versus C(18)) behave similarly. In contrast, defective and partly melted monolayers are more sensitive to stress than are their well-ordered analogues, and they are more prone to irreversible changes. In all cases, the intensity of methyl C-H stretching modes decreases with applied pressure, indicating a loss of net orientational order among the terminal methyl groups. The magnitude of this effect in well-ordered layers can be compared with the theoretical sensitivity of the resonant sum-frequency signal to molecular orientation. On these grounds, an increased population of terminal gauche conformers is identified as the disordering mechanism under pressure. PMID- 12720465 TI - Asymmetric alternating copolymerization of cyclohexene oxide and CO2 with dimeric zinc complexes. AB - Dimeric zinc complex 2a [ = Et(2)Zn(2)(1a)(2)] has been synthesized by the reaction of Et(2)Zn and (S)-diphenyl(pyrrolidin-2-yl)methanol (1a-H). X-ray crystallography revealed that the alkoxide ligand replaced one of the two ethyl groups of Et(2)Zn and formed a five-membered chelate ring through a Zn-N dative bond. Two zinc centers were bridged by oxygen atoms to form a Zn(2)O(2) four membered ring with a syn relationship between the two ethyl groups on the zinc centers. Dimeric zinc complex 2a was an active catalyst for asymmetric alternating copolymerization of cyclohexene oxide and CO(2). An MALDI-TOF mass spectrum of the obtained copolymer showed that the copolymerization was initiated by the insertion of CO(2) into Zn-alkoxide to give [(S)-diphenyl(pyrroridin-2 ly)methoxy]-[C(=O)O-(1,2-cyclohexylene)-O](n)-H (copolymer I), including chiral ligand 1a as an initiating group. Complex 3a-OEt ( = EtZn(1a)(2)ZnOEt), in which an ethoxy group replaced one of the two ethyl groups in 2a, also polymerized cyclohexene oxide and CO(2) with higher catalytic activity and enantioselectivity than 2a and afforded EtO-[C(=O)O-(1,2-cyclohexylene)-O](n)-H ( = copolymer III), including an ethoxy group as an initiating group. Throughout the studies, dimeric zinc species are indicated to be the active species for the copolymerization. It is also depicted that the substituent on the aryl moiety in diaryl(pyrrolidin-2 yl)methanol 2b-e influenced the polymerization activity. PMID- 12720466 TI - Thermal and electrochemical C-X activation (X = Cl, Br, I) by the strong Lewis acid Pd3(dppm)3(CO)2+ cluster and its catalytic applications. AB - The stoichiometric and catalytic activations of alkyl halides and acid chlorides by the unsatured Pd(3)(dppm)(3)(CO)(2+) cluster (Pd(3)(2+)) are investigated in detail. A series of alkyl halides (R-X; R = t-Bu, Et, Pr, Bu, allyl; X = Cl, Br, I) react slowly with Pd(3)(2+) to form the corresponding Pd(3)(X)(+) adduct and "R(+)". This activation can proceed much faster if it is electrochemically induced via the formation of the paramagnetic species Pd(3)(+). The latter is the first confidently identified paramagnetic Pd cluster. The kinetic constants extracted from the evolution of the UV-vis spectra for the thermal activation, as well as the amount of electricity to bring the activation to completion for the electrochemically induced reactions, correlate the relative C-X bond strength and the steric factors. The highly reactive "R(+)" species has been trapped using phenol to afford the corresponding ether. On the other hand, the acid chlorides react rapidly with Pd(3)(2+) where no induction is necessary. The analysis of the cyclic voltammograms (CV) establishes that a dissociative mechanism operates (RCOCl --> RCO(+) + Cl(-); R = t-Bu, Ph) prior to Cl(-) scavenging by the Pd(3)(2+) species. For the other acid chlorides (R = n-C(6)H(13), Me(2)CH, Et, Me, Pr), a second associative process (Pd(3)(2+) + RCOCl --> Pd(3)(2+.....)Cl(CO)(R)) is seen. Addition of Cu(NCMe)(4)(+) or Ag(+) leads to the abstraction of Cl(-) from Pd(3)(Cl)(+) to form Pd(3)(2+) and the insoluble MCl materials (M = Cu, Ag) allowing to regenerate the starting unsaturated cluster, where the precipitation of MX drives the reaction. By using a copper anode, the quasi-quantitative catalytic generation of the acylium ion ("RCO(+)") operates cleanly and rapidly. The trapping of "RCO(+)" with PF(6)(-) or BF(4)(-) leads to the corresponding acid fluorides and, with an alcohol (R'OH), to the corresponding ester catalytically, under mild conditions. Attempts were made to trap the key intermediates "Pd(3)(Cl)(+)...M(+)" (M(+) = Cu(+), Ag(+)), which was successfully performed for Pd(3)(ClAg)(2+), as characterized by (31)P NMR, IR, and FAB mass spectrometry. During the course of this investigation, the rare case of PF(6)(-) hydrolysis has been observed, where the product PF(2)O(2)(-) anion is observed in the complex Pd(3)(PF(2)O(2))(+), where the substrate is well-located inside the cavity formed by the dppm-Ph groups above the unsatured face of the Pd(3)(2+) center. This work shows that Pd(3)(2+) is a stronger Lewis acid in CH(2)Cl(2) and THF than AlCl(3), Ag(+), Cu(+), and Tl(+). PMID- 12720467 TI - Alcoholysis of acylpalladium(II) complexes relevant to the alternating copolymerization of ethene and carbon monoxide and the alkoxycarbonylation of alkenes: the importance of Cis-coordinating phosphines. AB - The mechanism and kinetics of the solvolysis of complexes of the type [(L L)Pd(C(O)CH(3))(S)](+)[CF(3)SO(3)](-) (L-L = diphosphine ligand, S = solvent, CO, or donor atom in the ligand backbone) was studied by NMR and UV-vis spectroscopy with the use of the ligands a-j: SPANphos (a), dtbpf (b), Xantphos (c), dippf (d), DPEphos (e), dtbpx (f), dppf (g), dppp (h), calix-6-diphosphite (j). Acetyl palladium complexes containing trans-coordinating ligands that resist cis coordination (SPANphos, dtbpf) showed no methanolysis. Trans complexes that can undergo isomerization to the cis analogue (Xantphos, dippf, DPEphos) showed methanolyis of the acyl group at a moderate rate. The reaction of [trans (DPEphos)Pd(C(O)CH(3))](+)[CF(3)SO(3)](-) (2e) with methanol shows a large negative entropy of activation. Cis complexes underwent competing decarbonylation and methanolysis with the exception of 2j, [cis-(calix diphosphite)Pd(C(O)CH(3))(CD(3)OD)](+)[CF(3)SO(3)](-). The calix-6-diphosphite complex showed a large positive entropy of activation. It is concluded that ester elimination from acylpalladium complexes with alcohols requires cis geometry of the acyl group and coordinating alcohol. The reductive elimination of methyl acetate is described as a migratory elimination or a 1,2-shift of the alkoxy group from palladium to the acyl carbon atom. Cis complexes with bulky ligands such as dtbpx undergo an extremely fast methanolysis. An increasing steric bulk of the ligand favors the formation of methyl propanoate relative to the insertion of ethene leading to formation of oligomers or polymers in the catalytic reaction of ethene, carbon monoxide, and methanol. PMID- 12720468 TI - The Rh4(CO)12-catalyzed hydroformylation of 3,3-dimethylbut-1-ene promoted with HMn(CO)5. Bimetallic catalytic binuclear elimination as an origin for synergism in homogeneous catalysis. AB - The bimetallic origins of catalytic synergism were studied using unmodified rhodium and manganese carbonyls as catalyst precursors during the low-temperature hydroformylation of 3,3-dimethylbut-1-ene to 4,4-dimethylpentanal in n-hexane solvent (T approximately 298 K, P(CO) = 1.0-4.0 MPa, P(H2) = 0.5-2.0 MPa). A dramatic increase in the catalytic rate was observed in the experiments conducted when both metals were used simultaneously. Detailed in-situ FTIR measurements indicated the observable presence of only homometallic complexes during catalysis, e.g., RCORh(CO)(4), Rh(4)(CO)(12), Rh(6)(CO)(16), HMn(CO)(5), and Mn(2)(CO)(10). The kinetics of product formation show a distinct linear-bilinear form in observables: k(1)[RCORh(CO)(4)][CO](-1)[H(2)] + k(2)[RCORh(CO)(4)][HMn(CO)(5)][CO](-1.5). The first term represents the classic unicyclic rhodium catalysis, while the second indicates a hydride attack on an acyl species. These spectroscopic and kinetic results strongly suggest that the origin of synergism is the presence of bimetallic catalytic binuclear elimination and not cluster catalysis. This appears to be the first detailed evidence for such a catalytic mechanism, and its implications for selectivity and nonlinear catalytic activity are accordingly discussed. PMID- 12720469 TI - Solution versus solid-state structure of ytterbium heterobimetallic catalysts. AB - The solution structures of the ytterbium heterobimetallic complexes Na(3)[Yb((S) BINOL)(3)] (1), K(3)[Yb((S)-BINOL)(3)] (2), and Li(3)[Yb((S)-BINOL)(3)] (3), belonging to a family of well-known enantioselective catalysts, are studied by means of NMR and circular dichroism (CD) in the UV and near-IR regions. The experimental NMR paramagnetic shifts were employed to obtain a refined solution structure of 1. NMR analysis demonstrated that 1, 2, and 3 have the same solution geometry but different magnetic susceptibility anisotropy D factors. By comparing XRD and NMR structures of 1, we demonstrate that, upon dissolution, this complex experiences a rearrangement from the crystalline C(3) symmetry into the solution D(3) symmetry. Remarkably, Yb is not bound to water in solution, and Ln-BINOL bonds are labile as demonstrated through EXSY. NIR-CD is confirmed especially sensitive to changes in the ytterbium coordination sphere. PMID- 12720470 TI - Adsorption morphology, light absorption, and sensitization yields for squaraine dyes on SnS2 surfaces. AB - The sensitization properties of two squaraine dyes adsorbed onto the van der Waals surface of n-doped tin disulfide single crystals were studied using atomic force microscopy (AFM), vis-NIR absorption spectroscopy, and photoelectrochemical techniques. Quantum yield per absorbed photon (QYAP) values of near unity were observed for submonolayer coverages of 2,4-bis(4-(N-methyl-N hexylamino)phenyl)squaraine (1-6SQ) in aqueous electrolyte when a sufficiently positive bias was applied, demonstrating the advantages of SnS(2) as a photoanode for fundamental studies of dye sensitization. Islandlike and microcrystalline morphologies, associated with aggregate formation and revealed by AFM, could be correlated with spectral shifts in both the absorbance and photoaction spectra. A related dye, 2,4-bis(4-(N,N-dimethyl)-2-hydroxyphenyl)squaraine (1-1OHSQ), at similar coverages showed slightly lower QYAP, ascribed to a recombination path due to the different aggregate structures. PMID- 12720471 TI - Scratching the surface of buckminsterfullerene: the barriers for Stone-Wales transformation through symmetric and asymmetric transition states. AB - General-gradient approximation (PBE) and hybrid Hartree-Fock density functional theories (B3LYP) in conjunction with basis sets of up to polarized triple-zeta quality have been applied to study the Stone-Wales transformation of buckminsterfullerene (BF) to yield a C(60) isomer of C(2)(v) symmetry with two adjacent pentagons (#1809). In agreement with earlier investigations, two different transition states and reaction pathways could be identified for the rearrangement from BF to C(60)-C(2)(v) on the C(60) potential energy surface (PES). One has C(2) molecular point group symmetry with the two migrating carbon atoms remaining close to the fullerene surface. The other one has a high-energy carbene-like (sp(3)) structure where a single carbon atom is significantly moved away from the C(60) surface. The carbene intermediate and the second transition state along the stepwise reaction path characterized previously at lower levels of theory do not exist as stationary points with the density functionals utilized here. The classical barriers of both mechanisms are essentially identical, 6.9 eV using PBE and 7.3 eV with B3LYP. PMID- 12720474 TI - Toward an understanding of bipolar disorder and its origin. AB - Bipolar disorder, a disease with significant morbidity and mortality, continues to present diagnostic and clinical challenges. Lifetime prevalence of bipolar I disorder has been estimated at 1.3%, with an equal distribution between males and females. Recognition of the illness may take years, but most patients are diagnosed before the age of 30. The role of genetic influences in bipolar disease is supported by family studies and high concordance rates among monozygotic twins. Current speculation proposes a likely interaction between genetic predisposition and environmental influences, including stressful life events. Diagnostic work-up should exclude mania secondary to drug use or general medical causes, particularly in patients whose symptoms begin after age 30 and in those with no family history of bipolar or unipolar disease. Patients with cyclothymia or thyroid dysfunction or postpartum women are at particular risk for bipolar disease. Substance abuse, which is extremely common among patients with bipolar disorder, interferes with diagnosis and can worsen the course of the disease. Alcohol dependence has been reported in approximately one third of those with bipolar I and one fifth of those with bipolar II disorder. To facilitate early diagnosis and effective management, clinicians should be aware of the risk factors, possible causes, and comorbidities of bipolar disease. PMID- 12720475 TI - Treatment of bipolar disorder. AB - Management strategies for bipolar disorder often entail relatively long-term, complex medication regimens that combine primary mood stabilizers, antipsychotic agents, antidepressants, and other medications, such as benzodiazepines. New strategies for the management of bipolar disorder have recently been evaluated in controlled clinical trials, including using newer anticonvulsants, replacing conventional antipsychotics with atypical antipsychotics, and using novel combination treatments. This article provides an overview of current management strategies for patients with bipolar disorder and describes how these approaches can be incorporated into clinical practice. PMID- 12720476 TI - Response, remission, and recovery in bipolar disorders: what are the realistic treatment goals? AB - Bipolar disorder presents particular challenges with regard to assessing response to therapy. Criteria for determining remission and recovery have been suggested for mood disorders, but the clinical usefulness of these terms in bipolar disorder is elusive. Formal psychological rating scales may be impractical in a routine medical practice setting. As an alternative, clinicians might probe for information about particular "signal events," such as sleep disturbances, that may herald mood fluctuations. The ultimate goal of bipolar management should be complete and sustained remission, whenever possible, although most patients will not achieve this status for any significant length of time. Furthermore, overaggressive management might entail pushing medication doses to intolerable levels. Individual treatment goals should always take into account patient acceptance of side effect burden, allowing for trade-offs between treatment effect and quality of life. Noncompliance with therapy, notoriously common among patients suffering from bipolar disorder, can stem from drug side effects, treatment ineffectiveness, or even treatment success if the patient misses the manic symptoms. Despite effective treatment, relapse is common. Realistic treatment goals should strive for sustained symptom abatement while maximizing patient quality of life from visit to visit. PMID- 12720477 TI - Bipolar disorder and schizophrenia: distinct illnesses or a continuum? AB - Bipolar disorder continues to present complex diagnostic and therapeutic challenges. Originally considered 2 separate diseases (mania and depression), bipolar disorder is now recognized to be a single disorder characterized by different subtypes and degrees of severity. Despite the availability of official guidelines, such as the DSM-IV and ICD-10, diagnosis is still problematic. Traditionally, bipolar disorder has been considered a clinical entity distinct from schizophrenia, although that assumption is being increasingly challenged. Proponents of a bipolar continuum theory support the concept of an expanded psychiatric continuum ranging from unipolar to bipolar disorders all the way to schizophrenia. This notion is supported by various independent findings. Both bipolar disorder and schizophrenia demonstrate a high degree of genetic transmissibility. Some data reported in family and twin studies suggest hereditary overlap between the 2 disorders. Gene mapping for both diseases is in its early stages, but certain susceptibility markers appear to be located on the same chromosomes. Bipolar disorder and schizophrenia also demonstrate some similarities in neurotransmitter dysfunction. As further indirect evidence of a possible association, many newer atypical antipsychotic agents approved for the treatment of schizophrenia are also proving useful for bipolar disorder. Ongoing research should aid in the understanding of bipolar disorder and foster the development of more effective treatment. PMID- 12720479 TI - The use of mood stabilizers as plasticity enhancers in the treatment of neuropsychiatric disorders. AB - Mood disorders have traditionally been conceptualized as neurochemical disorders, but there is now evidence from a variety of sources demonstrating regional reductions in central nervous system (CNS) volume, as well as reductions in the numbers and/or sizes of glia and neurons in discrete brain areas. Although the precise cellular mechanisms underlying these morphometric changes remain to be fully elucidated, the data suggest that severe mood disorders are associated with impairments of structural plasticity and cellular resilience. It is thus noteworthy that lithium and valproate have recently been demonstrated to robustly increase the expression of the cytoprotective protein bcl-2 (an abbreviation for the B-cell lymphoma/leukemia-2 gene) in the CNS in vivo and in cells of human neuronal origin. Lithium and valproate also robustly activate a signaling cascade utilized by endogenous growth factors-the extracellular signal-regulated kinase (ERK) mitogen-activated protein (MAP) kinase pathway. Complementary human studies have shown that chronic lithium administration significantly increases gray matter content in a regionally selective manner, suggesting a reversal of illness related atrophy and an increase in the volume of the neuropil. These unique and unexpected properties of lithium and valproate suggest that they may have broader utility as adjunctive agents in the treatment of a variety of neuropsychiatric disorders associated with cell atrophy or loss. The adjunctive use of these agents-at low doses-may provide the trophic support necessary to restore, enhance, and maintain normal synaptic connectivity, thereby allowing the chemical signal to reinstate the optimal functioning of critical circuits necessary for normal functioning. PMID- 12720480 TI - Rationale for using lithium in combination with other mood stabilizers in the management of bipolar disorder. AB - Bipolar disorder is a complex illness, and no single agent has been proven in randomized, placebo-controlled trials to effectively prevent and/or control all aspects of the illness-acute mania, rapid cycling, and breakthrough depression. However, for the most important issue, prophylaxis of episodes, lithium has more evidence of efficacy than any other agent. Like lithium, typical antipsychotics, carbamazepine, divalproex, and the atypical antipsychotic olanzapine are effective in the treatment of mania. Carbamazepine, divalproex, and olanzapine seem effective in preventing manic episodes but, like lithium, are less effective in preventing depression. Few trials have been conducted in the more difficult-to treat characteristics of bipolar disorder, specifically, rapid cycling and break through depression. For patients with rapid cycling, carbamazepine or divalproex therapy may improve symptoms, but only lamotrigine has been shown to reduce cycling, mostly in the bipolar II group, in a randomized, placebo-controlled study. For the treatment of depressive episodes, lithium and olanzapine have shown modest efficacy in controlled trials, and among the mood stabilizers, lamotrigine has the most robust effect. Because manic symptoms may respond best to one agent and depressive symptoms to another, combination therapy may be the optimal treatment for many patients with bipolar disorder. For example, lithium augmentation may improve overall response rates to treatment with carbamazepine or divalproex, and the lithium-lamotrigine combination should provide effective prevention of both mania and depression. Also, each mood stabilizer may be given at lower doses when given in combination, resulting in a reduced side effect burden and improved compliance. PMID- 12720481 TI - Drug combinations for mania. AB - With the release of new medications into the armamentarium for the treatment of bipolar disorder, clinicians are required to make prudent treatment decisions in light of insufficient research data for patients with a difficult-to-control illness. Increasingly, clinicians are turning toward a combination or adjunctive treatment out of necessity. Many studies suggest effectiveness of add-on agents in patients with mania who are unresponsive to one or more drugs, while only a limited number of controlled trials actually compare one particular combination regimen to another. Despite this lack of data, there has been no lack of advice for the clinician from clinical recommendations in the form of expert treatment guidelines to case reports describing suggestive findings from the off-label use of newer agents. With a particular emphasis on the treatment of mania, this article reviews the clinical data on the individual agents of foreseeable use in combination treatment for bipolar disorder. We suggest beginning with an agent of established effectiveness when combining medications for the treatment of bipolar disorder. PMID- 12720482 TI - Lithium combinations in acute and maintenance treatment of unipolar and bipolar depression. AB - Bipolar illness and unipolar depression are both affective disorders associated with high lifetime morbidity and premature mortality due to suicide. Numerous double-blind, placebo-controlled trials have shown that lithium augmentation therapy is effective in treating acute episodes of bipolar depression, refractory major depression, and delusional depression as well as in reducing recurrences of these illnesses. Lithium is the only agent approved by the U.S. Food and Drug Administration for maintenance treatment of bipolar disorder. Further research is needed to specifically address whether the antidepressant effect of adding lithium is greater in bipolar disorder or in unipolar depressions. This article will summarize available evidence and clinical considerations regarding the use of lithium augmentation in acute and maintenance treatment of unipolar and bipolar depressions. PMID- 12720483 TI - Drug interactions of lithium and other antimanic/mood-stabilizing medications. AB - Treatments for bipolar disorder often consist of a combination of 2 or more medications that have mood-stabilizing properties. Lithium is the only agent that has shown efficacy for the treatment of acute mania and acute depression as well as for the prevention of recurrence of mania and depression in patients with bipolar disorder. Other agents frequently used as mood stabilizers in monotherapy and in combination with lithium are valproate and carbamazepine. Several other drugs that have mood-stabilizing properties are used in many combinations to effectively manage the spectrum of symptoms of bipolar disorder throughout the acute and maintenance phases. However, side effects and drug-interaction precautions accompany all agents used to treat bipolar disorder, and often it can be difficult to determine whether a patient is suffering from side effects of treatment or symptoms of the illness itself. It is important to use the lowest effective dose of any treatment medication, to monitor laboratory values closely, and to be familiar with the side effects and interaction precautions of each medication that a patient is taking. Successful management of medications for the patient with bipolar disorder is a challenge, but increased effort and vigilance by clinicians mean a reduced risk of hospitalization and fewer office visits for patients. PMID- 12720484 TI - Lithium treatment and suicide risk in major affective disorders: update and new findings. AB - BACKGROUND: Evidence that therapeutic benefits of psychiatric treatments include reduction of suicide risk is remarkably limited and poorly studied. An exception is growing evidence for such suicidal risk reduction with long-term lithium maintenance. This report updates and extends analyses of lithium treatment and suicides and attempts. METHOD: We pooled data from studies providing data on suicidal acts, patients at risk, and average exposure times with or without lithium maintenance therapy, and considered effects of lithium on selected subgroups. RESULTS: Data from 34 reported studies involved 42 groups with lithium maintenance averaging 3.36 years, and 25 groups without lithium followed for 5.88 years, representing 16,221 patients in a total experience of 64,233 person-years. Risks for all suicidal acts/100 person-years averaged 3.10 without lithium versus 0.210 during treatment (93% difference) versus approximately 0.315 for the general population. For attempts, corresponding rates were 4.65 versus 0.312 (93% difference), and for completed suicides, 0.942 versus 0.174 (82% difference). Subjects with bipolar versus various recurrent major affective disorders showed similar benefits (95% vs. 91% sparing of all suicidal acts). Risk reductions for unipolar depressive, bipolar II, and bipolar I cases ranked 100%, 82%, and 67%. Suicide risk without lithium tended to increase from 1970 to 2002, with no loss of effectiveness of lithium treatment. CONCLUSION: The findings indicate major reductions of suicidal risks (attempts > suicides) with lithium maintenance therapy in unipolar >/= bipolar II >/= bipolar I disorder, to overall levels close to general population rates. These major benefits in syndromes mainly involving depression encourage evaluation of other treatments aimed at reducing mortality in the depressive and mixed phases of bipolar disorder and in unipolar major depression. PMID- 12720485 TI - Selecting effective long-term treatment for bipolar patients: monotherapy and combinations. AB - This article explores the roles of monotherapy and drug combinations in finding effective long-term treatment for individual patients with bipolar disorder. While current practice relies heavily on combinations, many bipolar patients can be successfully stabilized if the initial monotherapy is carefully selected according to the patient's clinical characteristics. The data show that (1) unequivocal responders to long-term monotherapies such as lithium, lamotrigine, or atypical neuroleptics each have a very different clinical profile, including clinical presentation and course, comorbidity, and, in particular, family history and (2) bipolar patients who respond very well to a long-term monotherapy have often completely failed on other monotherapies. Combinations appear indicated particularly in bipolar patients who are treatment-resistant to monotherapy, do not tolerate it well, or have not yet exhibited the clinical characteristics needed to choose an effective monotherapy. PMID- 12720486 TI - The therapeutic potential of novel cardiotonic agents. AB - During the course of treatment of heart failure patients, cardiotonic agents are inevitable for improvement of myocardial dysfunction. Clinically available agents, such as beta-adrenoceptor agonists and selective phosphodiesterase 3 inhibitors, act mainly via cyclic AMP/protein kinase A-mediated facilitation of Ca(2+) mobilisation (upstream mechanism). These agents are associated with the risk of Ca(2+) overload leading to arrhythmias, myocardial cell injury and premature cell death. In addition, they are energetically disadvantageous because of an increase in activation energy and metabolic effects. Cardiac glycosides act also via an upstream mechanism and readily elicit Ca(2+) overload with a narrow safety margin. No currently available agents act primarily via an increase in the myofilament sensitivity to Ca(2+) ions (central and/or downstream mechanisms). Novel Ca(2+) sensitisers under basic research may deserve clinical trials to examine the therapeutic potential to replace currently employed agents in acute and chronic heart failure patients. Molecular mechanisms of action of Ca(2+) sensitisers are divergent. In addition, they show a wide range of discrete pharmacological profiles due to additional actions associated with individual compounds. Therefore, the outcome of clinical trials has to be explained carefully based on these mechanisms of actions. PMID- 12720487 TI - New developments in the pharmacological treatment of chronic heart failure. AB - In recent years, rapid growth in the understanding of the pathophysiology of chronic heart failure has allowed for insights into many potential new therapeutic strategies. Yet until now, despite sound biological basis for efficacy and success in early-Phase studies, novel agents have not stood up to the scrutiny of late-Phase clinical trials. Indeed, remarkably negative results have been observed for vasopeptidase inhibitors, endothelin receptor antagonists and agents which block immune activation. However, efficacy data from other novel agents are still awaited, including the selective aldosterone receptor antagonist eplerenone, arginine vasopressin inhibitors, erythropoietin and hydroxy-methyl glutaryl coenzyme A reductase inhibitors. Other classes of drugs which may enter clinical development include cardiac metabolic agents, matrix metalloproteinase inhibitors and advanced glycation end product antagonists. That the mortality and morbidity of patients with chronic heart failure remain unacceptably high makes the ongoing commitment to exploration of new drug therapies for the condition critical. PMID- 12720488 TI - The therapeutic potential of bradykinin B2 receptor agonists in the treatment of cardiovascular disease. AB - The nonapeptide bradykinin (BK) is a Janus-faced hormone, which exerts pathophysiological as well as pronounced beneficial physiological effects, mainly by stimulation of BK B(2) receptors. In various animal models and in humans it has been shown that the stimulation of BK B(2) receptors is not only implicated in the pathogenesis of inflammation, pain and tissue injury but also in powerful cardioprotective mechanisms. Either exogenous administration of BK or locally increased BK concentrations as a consequence of the inhibition of its metabolic breakdown by angiotensin-converting enzyme inhibitors, reveal the significant contribution of BK in powerful cardioprotective mechanisms. These are mainly triggered by the synthesis and release of the vasorelaxant, anti-hypertrophic and anti-atherosclerotic endothelial mediators nitric oxide, prostaglandins and tissue-type plasminogen activator, by ischaemic preconditioning and by an increase in insulin sensitivity. Consequently, BK B(2) receptor agonists may have important clinical value in the treatment and prevention of various cardiovascular disorders such as hypertension, ischaemic heart disease, left ventricular hypertrophy, ventricular remodelling and congestive heart failure as well as diabetic disorders by mimicking the reported beneficial effects of BK. However, none of the currently known potent and selective peptide and non-peptide agonists of BK B(2) receptors--RMP-7 (lobradamil, Cereport; Alkermes), JMV-1116 (Fournier), FR-190997 (Fujisawa) and FR-191413 (Fujisawa)--have been selected for a clinical assessment in cardiovascular indications. One major challenge of this approach is the still unanswered question of whether there is a sufficient safe therapeutic window between potential cardioprotective and pro-inflammatory effects following BK B(2) receptor agonism. PMID- 12720489 TI - Therapeutic potential of active stent coating. AB - Various clinical studies have shown the superiority of stent implantation as compared to conventional balloon angioplasty for the treatment of significant coronary stenosis. However, restenosis remains a major drawback of this interventional technique. Against the background of this serious problem, the concept of stent coating has been developed. In general, coatings can be classified into two types: passive coatings, which only serve as a barrier between the stainless steel, and the tissue and active coatings, which directly interfere with the process of intima proliferation. At this moment, primarily immunosuppressive and cytostatic substances are used as active coatings. Large randomised studies have shown that this novel concept can be successfully implemented into clinical practice. Beside these promising results, studies also revealed potential risks of this new approach. Not only the dosage of the drug but also an optimised kinetic of drug release seem to be essential in preventing restenosis. As with most drugs, the inhibition of neointima proliferation is not restricted to vascular smooth muscle cells but also affects the process of re endothelialisation, thus we may face a new pitfall of late-stent thrombosis. Although this technique may harbour potential risks, the introduction of stent coating has the potential to dramatically reduce the incidence of restenosis and an exciting chapter in the field of cardiology has been opened. PMID- 12720490 TI - New antithrombotic drugs on the horizon. AB - Venous and arterial thromboembolism are a major cause for morbidity and mortality. The list of established drugs for the prevention of thrombus formation and embolisation includes heparins, hirudin and derivatives, aspirin, ADP and glycoprotein IIb/IIIa receptor antagonists, as well as vitamin K antagonists. Several limitations exist for these drugs that have stimulated the search for new and better anticoagulants. A series of selective clotting factor Xa inhibitors and direct factor IIa (thrombin) inhibitors are on the horizon, two of which are getting close to broad clinical application. Additional therapeutics that are still under preclinical and clinical investigation include inhibitors of the tissue factor pathway/factor VII complex, clotting factor VIII and XIII inhibitors and modulators of the protein C pathway or of endogenous fibrinolysis, as well as novel antiplatelet drugs. This review is focused on the current status of development of novel antithrombotics and their clinical potential. Even though only a few of a broad array of antithrombotic agents have reached clinical testing, some hold the potential for significant improvement in efficacy and safety of anticoagulant therapy. PMID- 12720492 TI - The role of 5-HT on the cardiovascular and renal systems and the clinical potential of 5-HT modulation. AB - The main peripheral sources of 5-hydroxytryptamine (5-HT) are as a neurotransmitter and local hormone in the gastrointestinal tract, and stored in circulating platelets and pulmonary neuroepithelial bodies. 5-HT has been shown to have many possible physiological and pathophysiological roles on the cardiovascular and renal systems. Thus, 5-HT may contribute to valvular heart disease, coronary artery disease, pulmonary hypertension, pulmonary embolism, pre eclampsia, peripheral vascular disease and diabetic nephropathy. Consequently, modulators of the 5-HT system have diverse clinical potential. For instance, selective 5-HT subtype 3 receptor (5-HT(3)) antagonists may have potential in the treatment of the pain associated with myocardial infarction. MCI-9042 (sarpogrelate) or other 5-HT(2A) antagonists may have clinical potential for the treatment of vasospastic angina, ischaemic heart disease, reperfusion injury and hindlimb ischaemia. Several modulators of 5-HT (5-HT transporter inhibitors, 5 HT(1B) and (2B) antagonists) may have potential alone or in combination in the treatment of pulmonary hypertension. In hypertension, agonists at the 5-HT(7) and antagonists at the 5-HT(2B) may reduce blood pressure, and in diabetes, sarpogrelate may protect against nephropathy. PMID- 12720491 TI - Factor X inhibitors. AB - Factor X plays a central role in coagulation, being the point of convergence of the extrinsic and intrinsic pathways of blood clotting. It may also act as one of the links between the coagulation and inflammatory pathways. These findings suggest that factor X may represent an attractive target for a new antithrombotic drug. Indeed, a factor X inhibitor, fondaparinux, has already been approved for clinical use to prevent post-operative deep vein thrombosis. Factor X inhibitors are also being evaluated for use in the treatment of the acute coronary syndromes, pulmonary embolism and deep vein thrombosis. Oral factor X inhibitors are also being developed, which may be of use in the outpatient prevention and/or treatment of stroke and thromboembolism. PMID- 12720493 TI - An update on the therapeutic potential of vitamin D analogues. AB - The review provides an evaluation of the therapeutic potential of vitamin D analogues in the context of the current understanding of vitamin D biochemistry, molecular biology and physiology. Vitamin D activity results from several circulating and intracellular physiological metabolites acting simultaneously through at least three receptors. Common analogues are reviewed. Although most vitamin D analogues have traditionally been analogues of 1,25-dihydroxyvitamin D, it may be better to deliver high doses of base vitamin or (analogues) of 25 hydroxyvitamin D. This would permit physiological endocrine, paracrine and autocrine vitamin D metabolism. Agonists or antagonists of tissue-specific vitamin D metabolic pathways could be coadministered. The importance of measuring endogenous vitamin D metabolites during in vivo studies and the pitfalls of extending data across species and time are emphasised. Human vitamin D analogue trials should include direct comparison against the related endogenous metabolite. PMID- 12720494 TI - The therapeutic potential of lactoferrin. AB - Lactoferrin (Lf), a natural defence iron-binding protein, is present in exocrine secretions that are commonly exposed to normal flora: milk, tears, nasal exudate, saliva, bronchial mucus, gastrointestinal fluids, cervicovaginal mucus and seminal fluid. Additionally, Lf is produced in polymorphonuclear leukocytes and is deposited by these circulating cells in septic sites. A principal function of Lf is that of scavenging non-protein-bound iron in body fluids and inflamed areas so as to suppress free radical-mediated damage and decrease accessibility of the metal to invading bacterial, fungal and neoplastic cells. Adequate sources of bovine and recombinant human Lf are now available for development of commercial applications. Among the latter are use of Lf in food preservation, fish farming, infant milk formula and oral hygiene. Other readily accessible body compartments for Lf administration include skin, throat and small intestine. Further research is needed for possible medicinal use in colon and systemic tissues. Although Lf is a natural product and should be highly biocompatible, possible hazards have been documented. PMID- 12720495 TI - Pemetrexed and its emerging role in the treatment of thoracic malignancies. AB - Pemetrexed (Alimta); Eli Lilly and Co., Indianapolis, IN, USA) is a unique multitargeted antifolate that inhibits at least three enzymes, thymidylate synthase, dihydrofolate reductase and glycinamide ribonucleotide formyltransferase. This novel drug is being evaluated in a comprehensive clinical programme for use in both front-line and second-line therapies. It has shown broad activity in a number of solid tumours, including colon cancer, breast cancer, lung cancer, head and neck, cervical cancer and others. While a number of antifolates have been evaluated in clinical trials, further development has been stopped or delayed by the occurrence of life-threatening toxicities. Similar trends were also initially observed with pemetrexed as well, but investigators later showed that these toxicities could be minimised with folic acid and vitamin B(12) supplementation included in the treatment regimen. Preliminary data indicate that this supplementation does not hamper drug efficacy in most tumour types and in many cases, supplemented patients exhibit improved clinical outcome. Here, the current data for pemetrexed in treating thoracic malignancies are reviewed, with special focus on malignant pleural mesothelioma and non-small cell lung cancer. PMID- 12720496 TI - The METHRO trials. AB - Venous thromboembolism is a common and potentially fatal complication among hospital in-patients, particularly those undergoing orthopaedic surgery. Current prophylactic strategies utilise low molecular weight heparins (LMWHs) and warfarin. However, painful subcutaneous injections for LMWHs and delays in achieving target anticoagulation for warfarin pose significant problems clinically. The Melagatran for THRombin inhibition in Orthopaedic surgery (METHRO) trial represents a landmark step in the sequential combination of subcutaneous and oral anticoagulation with melagatran and ximelagatran, respectively, for surgical venous thromboprophylaxis. These agents have proven to be as effective and safe as LMWHs. Furthermore, with no need for dosage adjustment or therapeutic drug monitoring there is emerging evidence that ximelagatran may replace warfarin as the anticoagulant of choice. PMID- 12720497 TI - Implications of the LIFE trial. AB - The recent Losartan Intervention For Endpoint Reduction in Hypertension (LIFE) study was conducted in patients with essential hypertension with electrocardiogram evidence of left ventricular hypertrophy. This showed that losartan compared to atenolol resulted in a significant reduction in the primary endpoint of cardiovascular morbidity and mortality, as well as a greater reduction in electrocardiographically-defined left ventricular hypertrophy. Importantly, this was despite a mean blood pressure reduction which was similar in both groups. Furthermore, the atenolol arm was associated with higher incidence of newly diagnosed diabetics. The LIFE study has firmly confirmed a place for losartan (and other angiotensin receptor blockers) in the management of hypertension. Losartan has also been shown to be effective in diabetics and in patients with atrial fibrillation, as well as in left ventricular hypertrophy regression. This trial also raises the possibility that beta-blockers should perhaps not be used as first-line monotherapy. PMID- 12720500 TI - Seizures as the presenting feature of rickets in an infant. PMID- 12720498 TI - What are the best first-line antihypertensives: answers and more questions from the ALLHAT study. AB - The Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial (ALLHAT) is considered the largest trial of antihypertensive therapy conducted to date. It compared the cardiovascular effects of three more recently developed classes of antihypertensive agents to those of a diuretic in an attempt to identify which of these drugs should be the recommended first-line agent for the treatment of hypertension. In this report the paper is discussed with the significance and the clinical implications of the results contrasted with existing evidence in hypertension research. Additionally, further questions raised by the ALLHAT study are explored with ongoing studies, and future directions with which to answer these emerging new questions are outlined. PMID- 12720501 TI - Staphylococcus aureus and stethoscopes. PMID- 12720502 TI - Self-experimentation" in vulnerable populations. PMID- 12720504 TI - Childhood obesity: modernity's scourge. PMID- 12720503 TI - Kava hepatotoxicity with Western herbal products: does it occur with traditional kava use? PMID- 12720505 TI - Children with autism deserve evidence-based intervention. PMID- 12720506 TI - Translating advances in schizophrenia treatment: a glass ceiling. PMID- 12720507 TI - Overweight and obesity in Australia: the 1999-2000 Australian Diabetes, Obesity and Lifestyle Study (AusDiab). AB - OBJECTIVE: To measure the prevalence of obesity in Australian adults and to examine the associations of obesity with socioeconomic and lifestyle factors. DESIGN: AusDiab, a cross-sectional study conducted between May 1999 and December 2000, involved participants from 42 randomly selected districts throughout Australia. PARTICIPANTS: Of 20,347 eligible people aged > or = 25 years who completed a household interview, 11,247 attended the physical examination at local survey sites (response rate, 55%). MAIN OUTCOME MEASURES: Overweight and obesity defined by body mass index (BMI; kg/m(2)) and waist circumference (cm); sociodemographic factors (including smoking, physical activity and television viewing time). RESULTS: The prevalence of overweight and obesity (BMI > or = 25.0 kg/m(2); waist circumference > 80.0 cm [women] or > or = 94.0 cm [men]) in both sexes was almost 60%, defined by either BMI or waist circumference. The prevalence of obesity was 2.5 times higher than in 1980. Using waist circumference, the prevalence of obesity was higher in women than men (34.1% v 26.8%; P < 0.01). Lower educational status, higher television viewing time and lower physical activity time were each strongly associated with obesity, with television viewing time showing a stronger relationship than physical activity time. CONCLUSIONS: The prevalence of obesity in Australia has more than doubled in the past 20 years. Strong positive associations between obesity and each of television viewing time and lower physical activity time confirm the influence of sedentary lifestyles on obesity, and underline the potential benefits of reducing sedentary behaviour, as well as increasing physical activity, to curb the obesity epidemic. PMID- 12720508 TI - Effect of warming adult diphtheria-tetanus vaccine on discomfort after injection: a randomised controlled trial. AB - OBJECTIVE: To determine whether warming or rubbing adult diphtheria tetanus (ADT) vaccine immediately before administration affects its temperature and reduces the incidence of pain. DESIGN: Double-blind, randomised controlled trial and in-vitro temperature study. SETTING: Emergency department (ED) of a regional hospital between April and December 2001. PATIENTS: Convenience sample of 150 patients aged 16 years or over who presented to the ED requiring ADT booster vaccination. INTERVENTION: Patients were randomised to receive vaccine that was "cold" (no deliberate warming), "rubbed" between the palms for 1 minute, or "warmed" in a 37 degrees C incubator; vaccine was administered as recommended in Australian guidelines. MAIN OUTCOME MEASURES: Incidence of pain and pain score on McGill Present Pain Intensity Questionnaire at 5 minutes, 24 hours and 48 hours after injection; and temperature of vaccine after preparation for simulated administration. RESULTS: The "cold" vaccine had significantly lower temperature (mean, 19.1 degrees C; 95% CI, 17.5-20.7 degrees C) than the "warmed" vaccine (mean, 28.9 degrees C; 95% CI, 28.4-29.4 degrees C) and "rubbed" vaccine (mean, 26.9 degrees C; 95% CI, 24.5-29.3 degrees C). There was no significant difference in incidence of pain between the groups who received vaccine prepared in different ways at any follow-up (5 min: P = 0.62; 24 h: P = 0.58; 48 h: P = 0.61) or overall (P = 0.99). Among those who completed follow-up, incidence of pain at any time was 77/138 (56%); there was no difference in their time-averaged pain scores (P = 0.63) or peak pain scores (P = 0.60). CONCLUSIONS: Warming or rubbing ADT vaccine does not reduce the incidence of pain after administration. Regardless of how ADT vaccine is prepared, its temperature approaches ambient by the time it is injected. PMID- 12720509 TI - Asthma symptoms associated with depression and lower quality of life: a population survey. AB - OBJECTIVE: To identify any association between asthma and depression and quality of life. DESIGN AND SETTING: A face-to-face Health Omnibus Survey of a random and representative sample of the South Australian population in August 1998. PARTICIPANTS: 3010 randomly selected participants aged 15 years and over. MAIN OUTCOME MEASURES: Prevalence of doctor-diagnosed asthma, and scores for depression (measured by PRIME-MD instrument) and quality of life (measured by SF 36) in affected participants. RESULTS: The prevalence of asthma was 9.9%. The prevalence of major depression was significantly higher for those who experienced dyspnoea, wakening at night with asthma, and morning symptoms of asthma. Quality of-life scores were also lower for the same groups. CONCLUSIONS: Depression is a serious but potentially remediable comorbidity with asthma that may affect appropriate diagnosis and outcome. PMID- 12720510 TI - Fatal fulminant hepatic failure induced by a natural therapy containing kava. AB - We describe a case of acute liver failure and death associated with the use of a preparation containing the "natural" anxiolytic kava (Piper methysticum) and passionflower (Passiflora incarnata). The patient died after a report by the Therapeutic Goods Administration (TGA) warning of the potential for hepatotoxicity associated with the use of kava-containing products. The general public and alternative medicine practitioners need to be aware of the potential for non-prescription drugs to cause serious hepatic reactions. PMID- 12720511 TI - The management of persistent pain. AB - Persistent pain is a complex mix of physical and psychological symptoms and is ideally managed by a biopsychosocial approach. Often the relative contributions of family and personal relationships, finances, work, past pain experiences and personality outweigh those of the nociceptive or neuropathic processes from which most pain originates. Recent advances in our understanding of the pathophysiology of pain may lead to improved drug treatments; however, non-drug treatments- education, lifestyle modification, exercise and reassurance--should be used routinely to improve patients' quality of life. Patients with persistent pain that is difficult to control or has complex psychosocial influences, or who have a history of medication misuse, should be referred to a multidisciplinary pain centre. Selected patients may be offered invasive options such as nerve blocks or spinal-cord stimulation. The best outcomes are achieved in patients treated in group-based pain-management programs using cognitive-behavioural therapy to improve physical function, change unhelpful thinking and improve patients' understanding of their situation. PMID- 12720512 TI - Are we ready for the Better Medication Management System? AB - The Better Medication Management System (BMMS) is an Australian project to provide electronic access to patient medication records. It aims to improve access to patient medication information and to reduce adverse drug events and hospital admissions. Evidence supporting the achievability of the BMMS aims is limited. Voluntary participation of patients in the BMMS may limit its ability to achieve its aims and capitalise on its potential benefits. PMID- 12720513 TI - Kava: herbal panacea or liver poison? AB - Following reports of liver toxicity, including liver failure, associated with extracts from the Pacific islands plant kava (Piper methysticum), these have been banned from sale as a herbal anxiolytic in many Western countries, to the detriment of Pacific island economies. Pacific Islanders have used kava extensively for centuries, without recognised liver toxicity. However, the population is small, and there has been no systematic evaluation of possible liver damage. For both economic and public health reasons, it is important to determine if kava is inherently hepatotoxic, and what the mechanisms of toxicity are. Such research could lead to safer kava extracts for sale in Western countries, or identification of a subpopulation who should not consume kava. PMID- 12720514 TI - A philosophical approach to rationing. AB - Rationing, the equitable allocation of medical resources, is both an economic and moral challenge--economic, because the various components of healthcare must be budgeted; moral, because the prioritisation of these resources is a value-laden decision. The moral debate about rationing pits individual choice against communal interests. The advocacy of equitable distribution of healthcare resources originates in arguments for distributive justice and a revised version of individual autonomy. If autonomy is defined strictly in terms of atomistic individuality, then the social obligations and duties of persons are subordinated to their individual rights. Alternatively, when people are defined by their relationships, "relational autonomy" balances responsibilities against the claims of individual rights to maximise distributive justice. The concept of relational autonomy provides medicine with a philosophical basis for communal rationing of healthcare resources. PMID- 12720515 TI - The clinical geneticist and the "new genetics". AB - The "new genetics" will provide new genetic tests that can be used for diagnosis, prognosis, treatment selection, carrier and predictive testing in affected families, and potentially for susceptibility testing for later-onset multifactorial disease and population screening. Doctors will increasingly need to consider the family implications of a genetic diagnosis--to identify family members at risk of the disorder or of having affected children and to consider how these individuals might be advised of their situation. Clinical geneticists can be a valuable resource for doctors who need advice about whether genetic testing is available, which tests to pursue, how to access testing services, and how to interpret and act on test results. Clinical geneticists also provide genetic counselling, a process which gives people understandable information about the genetic disorder in the family, and makes the information useful for decision-making given the person's unique circumstances and beliefs. The Internet will increasingly be a key source of information about genetic disorders for patients, their families and healthcare professionals. PMID- 12720516 TI - Prescriptions for antipsychotics in general practice. PMID- 12720517 TI - Hepatitis risk and vaccination among Australian travellers overseas. PMID- 12720519 TI - The New South Wales Medical Board policy on treating self and family. PMID- 12720518 TI - The decline in bulk-billing and increase in out-of-pocket costs for general practice consultations in rural areas of Australia, 1995-2001. PMID- 12720520 TI - Comprehensive care for people with schizophrenia living in the community. PMID- 12720521 TI - Reducing mental illness stigma and discrimination - everybody's business. AB - The stigma associated with schizophrenia is pervasive, both in the community and among healthcare workers, and forms a real barrier to optimal recovery from the illness. The negative consequences of stigma include discrimination in housing, education and employment, and increased feelings of hopelessness in people with schizophrenia. Health professionals have a responsibility to improve their own attitudes and behaviour towards people with schizophrenia so they do not contribute to the stigma. Educational campaigns aimed at people in the community and media personnel could help to demystify mental illness and reduce the portrayal of offensive stereotypes of people with schizophrenia. PMID- 12720522 TI - The configuration of mental health services to facilitate care for people with schizophrenia. AB - In Australia, the configuration of public mental health services varies between States and Territories, but, overall, community-based services are increasingly integrated and responsive to people with schizophrenia. Community-based services include mobile crisis teams, providing home-based acute treatment, and case management services for ongoing treatment. Service improvements have been uneven across Australia. Some people with schizophrenia in psychiatric crisis have had difficulty accessing either home-based acute psychiatric treatment or acute psychiatric beds. Social isolation and lack of meaningful occupation continue to be a problem for people with schizophrenia. Psychosocial interventions can enhance reintegration into the community. However, the number of community-based psychosocial rehabilitation programs is still inadequate. PMID- 12720523 TI - Overcoming barriers to reintegration of patients with schizophrenia: developing a best-practice model for discharge from specialist care. AB - Many people with schizophrenia are in regular contact with their general practitioners. GPs commonly play a sentinel role in management, but may require support from Area Mental Health Services (AMHSs). The CLIPP (Consultation and Liaison in Primary-care Psychiatry) shared-care model of patient management combines a collocated consultation/liaison service for managing referrals from GPs to specialists with a carefully structured approach to long-term care of patients transferred from AMHS care to GPs. The CLIPP model uses the concept of a "relapse signature", involving recognition of early warning signs of relapse, to simplify clinical monitoring of patients with schizophrenia. PMID- 12720524 TI - Pharmacological approaches to the management of schizophrenia. AB - Pharmacological treatment remains the mainstay of the management of schizophrenia. Older, "typical" antipsychotics carry a significant burden of side effects, notably extrapyramidal and neurocognitive side effects. Newer, "atypical" agents carry a lower risk of extrapyramidal side effects. They appear to have added benefit for treating negative and cognitive symptoms of schizophrenia, and hence can enhance the quality of life of some patients. The choice of particular agents for individual patients requires a balancing of efficacy and side effects. Medication is only one element of what should be an individualised comprehensive treatment plan for people with schizophrenia. PMID- 12720526 TI - Medical comorbidity in schizophrenia. AB - Schizophrenia has been described as a "life-shortening disease", and physical comorbidity accounts for 60% of premature deaths not related to suicide. People with schizophrenia and other mental illnesses have a higher rate of preventable risk factors such as smoking, high alcohol consumption, poor diet, and lack of exercise. Recognition and management of morbidity in people with mental illness are made more difficult by barriers related to the patient, the illness, the attitudes of medical practitioners, and the structure of healthcare delivery services. Improved detection and treatment of medical illness in people with schizophrenia will have significant benefits for their psychosocial functioning and overall quality of life. PMID- 12720525 TI - Managing patients with "treatment-resistant" schizophrenia. AB - Patients who fail to respond adequately to pharmacological treatment present an ongoing therapeutic challenge. The term "incomplete recovery" (IR) is preferred to the current term "treatment resistance" to describe these patients. IR should be considered from a multidimensional perspective that includes a broad range of symptoms and functional disabilities that are relevant to schizophrenia. The approach to the incompletely recovered patient needs to be systematic, with consideration given to the factors that may hamper recovery. "Atypical" (second generation) antipsychotic drugs target various domains of symptoms relevant to IR. Adjunctive treatment strategies (eg, mood stabilisers, antidepressants, combinations of antipsychotics) may be useful, but should be undertaken in specialist psychiatric settings. Although pharmacological treatment is a necessary first step in managing incompletely recovered patients, adjunctive psychosocial interventions are needed to optimise treatment success. PMID- 12720527 TI - Substance misuse in patients with schizophrenia: a primary care guide. AB - Smoking presents a substantial health and economic burden to people with schizophrenia. Comorbid use of other substances is common, under-recognised, and associated with a number of serious adverse consequences, such as psychotic relapse and poorer social outcomes. All patients with schizophrenia need to be screened for substance misuse. Effective interventions involve integrated, modified pharmacological and psychosocial strategies. PMID- 12720528 TI - A meaningful day: integrating psychosocial rehabilitation into community treatment of schizophrenia. AB - While many of the overt symptoms of schizophrenia may be controlled by medication, the associated psychiatric disability requires ongoing psychosocial rehabilitation and support in the community. The general practitioner can play a crucial role in this rehabilitative process, through encouraging good physical health in the person with schizophrenia; referring the person to a local day program and other psychiatric disability support services; educating the person about the diagnosis and practical self-help; involving families and referring them to support organisations and group therapy programs; and encouraging use of local community facilities and employment training, where appropriate. PMID- 12720529 TI - Prolonged lifespan by defective insulin signalling? PMID- 12720530 TI - Multi-center study on the characteristics and treatment strategies of patients with Graves' orbitopathy: the first European Group on Graves' Orbitopathy experience. AB - To improve management of patients with Graves' orbitopathy, a multi-center collaborative approach is necessary in order to have large enough sample sizes for meaningful randomized clinical trials. This is hampered by a lack of consensus on how to investigate the eye condition. The European Group on Graves' Orbitopathy aims to overcome this and has designed a preliminary case record form (CRF) to assess Graves' orbitopathy patients. This form was used in this first multi-center study. AIM: To investigate patient characteristics and treatment strategies in 152 new consecutively referred patients with thyroid eye disease seen in nine large European referral centers. METHODS: Newly referred patients with Graves' orbitopathy were included who were seen between September and December 2000. Demographic data and a complete ophthalmological assessment were recorded. RESULTS: One-hundred and fifty-two patients (77% females) were included. Diabetes was present in 9%, and glaucoma or cataract in 14% of patients. Forty percent were current smokers, 9% also had dermopathy, and only 33% reported a positive family history of thyroid disease. Mild eye disease was seen in 40%, moderately severe eye disease was seen in 33% and severe eye disease was seen in 28% of patients. Soft tissue involvement was the most frequent abnormality (seen in 75%), proptosis > or =21 mm was found in 63%, eye motility dysfunction in 49%, keratopathy in 16% and optic nerve involvement was found in 21% of patients. According to the clinical impression, 60% had active eye disease. Immunosuppressive treatment was planned more frequently in active patients (57/86; 66%) than in inactive patients (5/57, 9%; Chi-square 46.16; P<0.02). There were no important differences among the eight centers regarding the severity and the activity of their patients. CONCLUSIONS: In view of the large number of patients recruited in only 4 months, multi-center studies in the eight EUGOGO centers appear to be feasible. PMID- 12720531 TI - Thyroid consequences of the Chernobyl nuclear power station accident on the Turkish population. AB - OBJECTIVE: The Chernobyl accident caused widespread effects across Europe and huge areas were radiocontaminated. The major impact of the accident on human health was a sharp increase in childhood thyroid carcinoma and autoimmune thyroid diseases in exposed populations. The thyroidal effects of the Chernobyl accident have been investigated in most European countries, except Turkey. The aim of the current study was therefore to determine the thyroidal consequences of the Chernobyl nuclear power station accident in a selected Turkish population. DESIGN: This study was designed as a sectional, area study, between October 2000 and March 2001, in two different regions of Turkey. According to the data of the Turkish Atomic Energy Authority, the eastern part of the Black Sea region was the most radiocontaminated area in Turkey at the time of Chernobyl accident, while Middle Anatolia was not seriously affected. Thus, Rize city, which is located in the eastern Black Sea region, served as a study area, and 970 adolescents, living in this region, comprised our study group (group R). On the other hand, Beypazari, which is located in Middle Anatolia, was chosen as the control region, and 710 adolescents living in this location were enrolled into the study as controls (group B). METHODS: During the study, thyroid ultrasounds were performed in all subjects and thyroid volumes were calculated. World Health Organization and International Council for Control of Iodine Deficiency Disorders criteria were used for the determination of goiter. Thyroid fine-needle aspiration biopsy with ultrasound guidance was performed when a nodule was detected. Blood samples for thyroid function tests and thyroid autoantibodies, and urine samples for urinary iodine excretion were collected from all subjects. RESULTS: Thyroid function tests were similar in both groups, but thyroid volumes were found to be higher in group B (13.93+/-5.04 vs 17.66+/-5.58 ml; P<0.001). The prevalence of goiter was found to be 28.25% in group R and 61.95% in group B (P<0.001). Thyroid nodules were determined in 6.28% of subjects in group R and 4.22% of subjects in group B (P=0.065). No malignant lesions were found in either of the regions. Although the percentage of autoantibody-positive subjects did not differ between groups (21.25% in group R vs 18.72% in group B), the mean anti-thyroglobulin level was found to be higher in group R (63.25+/-378.60 vs 51.97+/-333.32 IU/ml; P<0.001) and the mean anti-thyroid peroxidase level was higher in group B (24.14+/-219.09 vs 48.82+/-568.50 IU/ml; P<0.001). The iodine status of the selected regions was found to be significantly different (median urinary iodine excretion was 131 microg/l in Rize and 54 microg/l in Beypazari). CONCLUSIONS: Although there was a slight increase in nodule prevalence and thyroid antibody positive subjects in the study group, it is hard to conclude that Turkey was affected by the Chernobyl accident. These results, at least the significant differences with regard to the prevalence of goiters between groups, may reflect the different iodine status of the selected regions. PMID- 12720532 TI - Ret/PTC activation does not influence clinical and pathological features of adult papillary thyroid carcinomas. AB - OBJECTIVE: RET proto-oncogene rearrangements (ret/PTCs) represent the most common genetic alterations found in papillary thyroid carcinomas (PTCs). Correlation of ret/PTC expression with clinical outcome is controversial. The aim of the present study was to analyze the frequency of RET rearrangements in adult PTCs, and to investigate if ret/PTCs influence biological behavior and clinical features of the cancers. DESIGN: Ret/PTC rearrangements were looked for in tIssue samples of 48 PTCs collected at our institution. Data about clinical and pathological features of the tumors were also reviewed. Three separate association analyses were carried out on the cohort evaluating the effects of, respectively, ret/PTC positivity, preferential RET tyrosine kinase domain (RET-TK) expression, and ret/PTC plus RET-TK positivity, on age, sex, tumor size, staging, number of neoplastic foci, and histological subtype. METHODS: The genetic study was conducted with the RT-PCR-Southern blot technique. Standard Student's t-test and Fisher exact test were applied for the association analyses. RESULTS: The molecular genetic study demonstrated the positivity of ret/PTC1 and ret/PTC3 in 13 of 48 tumors (27.1%), and an exclusive or preferential RET-TK expression in 17 cases (35.4%). None of the three genetico-clinical analyses showed any significant association between ret/PTC expression and the clinical and pathological features of the cancers. CONCLUSIONS: These data indicate that RET rearrangements may not play any distinctive role in driving histotype development and cancer progression in these neoplasms. Moreover, they weaken the possibility of using ret/PTC as a prognostic marker for papillary thyroid carcinomas. PMID- 12720533 TI - Treatment of GH-deficient children with two different GH doses: effect on final height and cost-benefit implications. AB - OBJECTIVE: Treatment of GH-deficient (GHD) children with higher doses of recombinant human GH (rhGH) than conventional ones has been reported to result in higher growth velocity and increased final height. These findings, however, were observed by comparing large but heterogeneous groups of children. We wanted to verify whether the same results could be obtained in two groups of appropriately well-matched children with isolated GHD treated with high vs conventional doses of rhGH. METHODS: Out of two cohorts of GHD children, cohort A (on a weekly rhGH dose of 0.3 mg/kg body weight) and cohort B (on a weekly rhGH dose of 0.15 mg/kg body weight), we selected two groups, each including 13 patients, who before treatment were matched for age, sex and height standard deviation score (SDS). They were followed up until final height. RESULTS: Final height SDS was significantly higher in group A (-0.45+/-0.36 (s.d.) vs -1.07+/-0.7; P=0.008), as well as height gain SDS (1.81+/-0.58 vs 1.23+/-0.62; P=0.002). The difference between final height SDS and target height SDS was positive only in group A and significantly higher in group A than in group B (0.33+/-0.51 vs -0.46+/-0.7; P=0.01). Glucose tolerance was always normal in the group treated with higher doses. CONCLUSION: The final height of children treated with higher doses of rhGH is increased, also in relation to their genetic target. The economical burden of this choice of treatment, however, has to be taken into account when evaluating the results. PMID- 12720534 TI - Regulation of CYP19 gene expression in primary human osteoblasts: effects of vitamin D and other treatments. AB - OBJECTIVE: Extragonadal estrogen biosynthesis is relevant for the regulation of bone metabolism. The aims of this paper were: (i) to examine CYP19 (aromatase) gene expression in primary cultures of osteoblasts under several hormone and cytokine treatments and (ii) to study the promoter usage of CYP19 in these cells. METHODS: Primary cultures of osteoblasts were obtained from healthy donors. The effects of vitamin D and other factors on CYP19 expression were analysed by semiquantitative RT-PCR. Furthermore, CYP19 alternative promoter usage under the different treatments was characterized by RT-PCR. RESULTS: CYP19 transcripts were detected in cultured human osteoblasts in serum-free conditions. Vitamin D, dexamethasone, 17beta-estradiol and testosterone increased transcript levels of CYP19, whereas interleukin-1beta or tumor necrosis factor alpha decreased them. Aromatase mRNA produced under treatment with vitamin D was transcribed from promoters I.4 and I.3, while stimulation with dexamethasone or dexamethasone plus vitamin D also involved promoter I.2. Testosterone activated promoters I.2 and I.4. CONCLUSIONS: Our results suggested that vitamin D, testosterone, estrogens and glucocorticoids regulate CYP19 gene expression in human primary osteoblasts and the main promoter used appears to be promoter I.4. Promoters pII and I.3 seem to be related to basal transcription and may mediate estrogen stimulation, while promoter I.2 seems to play a role in the effect of glucocorticoids. These findings indicate that vitamin D and several hormones regulate local estrogen synthesis in human osteoblasts mainly through usage of promoters I.4 and I.3. PMID- 12720535 TI - Effect of cyclical intermittent etidronate therapy on circulating osteoprotegerin levels in patients with rheumatoid arthritis. AB - OBJECTIVE: To evaluate the role of serum osteoprotegerin (OPG) as a biochemical marker for disease activity assessment and drug monitoring in patients with rheumatoid arthritis (RA) treated with cyclical etidronate. DESIGN: Forty patients (35 women and 5 men) with RA of <5 years duration were randomized to receive intermittent cyclical etidronate therapy in conjunction with anti rheumatic therapy or anti-rheumatic therapy alone (without etidronate) in a 2 year, open-label protocol. METHODS: Radiographs of hands and feet and serum samples for the determination of OPG, amino terminal propeptide (PINP), cross linked C-telopeptide (ICTP) and amino terminal telopeptid of type I collagen were obtained at baseline and at 24 months. RESULTS: Etidronate treatment had no effect on circulating OPG levels, although the significant decline in PINP and ICTP (P=0.001 and P=0.04 respectively) reflected the efficacy of the anti resorptive therapy. At baseline and at study termination, serum OPG correlated significantly with age (r=0.45; P=0.003 and r=0.56; P=0.0002 respectively). OPG was not related to biochemical markers of bone metabolism, indices of disease activity or radiographic disease progression. At baseline, the mean serum OPG was higher in patients receiving 5-10 mg/day prednisone (82.8+/-4.0 pg/ml, n=16) compared with those receiving <5 mg/day or with no prednisone (69.7+/-4.7 pg/ml, n=23) (P=0.05). CONCLUSIONS: Our results suggested that serum OPG measurement, perhaps because of the complexity of the regulation of the OPG, may be difficult to utilize in the evaluation of anti-resorptive therapy. Moreover, low dose corticosteroid-associated osteoporosis is probably not mediated by inhibition of OPG. PMID- 12720536 TI - Pitfalls in endosonographic imaging of suspected insulinomas: pancreatic nodules of unknown dignity. AB - OBJECTIVE: Endosonography enables localization and characterization of gastroenteropancreatic neuroendocrine tumors. We have studied the problem of misleading abnormalities of pancreatic morphology as obtained by endosonographic imaging. DESIGN AND METHODS: A total of 438 endosonographies performed for known or suspected diseases of the adrenal glands and/or the pancreas and/or suspected metastases in the neighboring tissues were analyzed. RESULTS: In the pancreas, nine benign insulinomas, four non-metastatic islet cell carcinomas, and multiple benign neuroendocrine tumors in one patient with multiple-endocrine neoplasia-1 (MEN-1) disease were detected and correctly localized as proven by postoperative histology. In three further patients with genetic diagnosis of MEN-1, asymptomatic tumors were detected and are under observation. However, we also found an 8 x 4 mm hypoechoic tumor in the cauda pancreatis of a patient with severe factitial hypoglycemia (glimepiride). In another patient suffering from severe hypoglycemia, a hypoechoic area of 24 x 10 mm in the processus uncinatus/caput pancreatis was found. Although organic hyperinsulinism was excluded, this patient underwent surgery because of suspected pancreatic carcinoma. There was normal pancreatic tissue in the abnormal region, which was also localized by intraoperative sonography. In a third patient with an adrenal carcinoma, a 6 x 3 mm hypoechoic nodule in the cauda pancreatis did not change its morphology over an observation period of 13 months, its clinical relevance is completely unclear. CONCLUSIONS: Pancreatic nodules of unknown dignity were detected in nearly 1% of our patients and must be considered to be a diagnostic problem. These experiences clearly show, on the one hand, that pancreatic endosonography is a very useful diagnostic support in the management of endocrine tumor patients. However, on the other hand, endosonography of endocrine organs is not a substitute for careful endocrinological examination and testing and must be considered in the context of endocrinological findings. PMID- 12720537 TI - Association between measures of insulin sensitivity and circulating levels of interleukin-8, interleukin-6 and tumor necrosis factor-alpha. Effect of weight loss in obese men. AB - OBJECTIVE: To study the association between anthropometric and metabolic parameters as well as the effect of weight loss on plasma levels of the adipose tissue-derived cytokines interleukin-8 (IL-8), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) in abdominal obese men. SUBJECTS: Nineteen obese (mean body mass index (BMI): 38.6+/-0.6 kg/m(2)) and ten lean men (mean BMI: 23.4+/-0.4 kg/m(2)) were included in the study. The obese subjects received a 4.2 MJ/day diet for 8 weeks, followed by 8 weeks on energy restriction (6.2 MJ/day) and 8 weeks on a weight-maintenance diet. MEASUREMENTS: A dual energy X ray absorptiometry (DEXA)-scan was performed to estimate body composition. Plasma levels of IL-8, IL-6 and TNF-alpha were measured by a specific ELISA method. Insulin sensitivity was assessed by the homeostasis model assessment method (HOMA). RESULTS: Plasma levels of IL-8 and IL-6 were 30-40% higher in obese as compared with lean subjects (P<0.05), whereas no group difference in TNF-alpha was observed. During the intervention, obese subjects obtained a 30% reduction in fat mass (P<0.001), fasting insulin (P<0.05) and HOMA (P<0.05). Plasma levels of TNF-alpha and IL-6 were decreased by 25-30% (P<0.001) but IL-8 was increased by 30% (P<0.001) after weight loss. IL-8 and IL-6 were correlated with measures of insulin resistance, and changes in IL-6 but not IL-8 were correlated with the improvement in insulin sensitivity after weight loss. CONCLUSION: Plasma levels of IL-8 and IL-6 were found to be increased and were correlated with measures of insulin resistance in abdominal obese male subjects. Weight loss was associated with changes in the circulating levels of IL-8, IL-6 and TNF-alpha indicating that these cytokines are influenced by weight loss. PMID- 12720538 TI - Reduced expression of uncoupling proteins-2 and -3 in adipose tissue in post obese patients submitted to biliopancreatic diversion. AB - OBJECTIVE: Little is known about the physiological role and the regulation of uncoupling proteins-2 and -3 (UCP-2 and -3) in adipose tissue. We investigated whether the expression of UCP-2 and -3 in adipose tissue was affected by weight loss due to a biliopancreatic diversion (BPD) and related to the daily energy expenditure (24-h EE). DESIGN: Ten morbidly obese subjects (mean body mass index +/- s.e.m.=49.80 +/- 2.51 kg/m(2)) were studied before and 18+/-2 Months after BPD. METHODS: We determined body composition using tritiated water and 24-h EE in a respiratory chamber. Adipose tissue UCP-2 and -3 mRNA, plasma insulin, glucose, free fatty acids (NEFA), free triiodothyronine (FT3), free thyroxine (FT4) and leptin were assayed before and after BPD. RESULTS: BPD treatment resulted in a marked weight loss (P<0.001) mainly due to a fat mass reduction. A significant decrease in 24-h EE/fat-free mass (FFM) (P<0.05) and in UCP-2 (P<0.05) and UCP-3 (P<0.05) mRNA was observed. A significant reduction in plasma insulin, glucose, NEFA, FT3, FT4 and leptin was seen after BPD. The decline in plasma leptin and FFA was tightly correlated with the decrease in both UCP-2 and -3. A significant correlation was found between changes in FT3 and variations in 24-h EE (r=0.64, P<0.05). In a multiple-regression analysis changes in 24-h EE/FFM after BPD were significantly correlated with changes in UCP-3 expression (P<0.05). CONCLUSION: These findings suggest that UCPs in adipose tissue may play a role in the reduction in 24-h EE observed in post-obese individuals. PMID- 12720539 TI - Sex steroids and body composition in men with cystic fibrosis. AB - OBJECTIVE: Delayed sexual maturation and low body weight is common in cystic fibrosis (CF). Concomitant data on sex hormones and concomitant body composition are lacking in men with CF. DESIGN: Cross-sectional study. SUBJECTS AND METHODS: Serum levels of testosterone, 17beta-oestradiol (E(2)), 25-hydroxyvitamin D (25(OH)D), sex hormone-binding globulin (SHBG) and LH were measured by RIA and total and regional lean body mass (LBM), fat body mass (FBM), bone mineral content and bone mineral density (BMD) were assessed by dual-energy X-ray absorptiometry, in men with CF (n=40; age 24.7+/-5.4 years) and age-matched healthy controls (n=28; age 25.7+/-3.7). Only men without acute disease exacerbation or systemic glucocorticoid treatment were included. RESULTS: Mean levels of hormonal serum parameters differed significantly between healthy controls (testosterone=20.2+/-5.5 nmol/l; E(2)=95.0+/-20.2 pmol/l; 25(OH)D=62.8+/ 28.3 nmol/l) and patients (testosterone=15.9+/-4.1 nmol/l; E(2)=60.7+/-19.4 pmol/l; 25(OH)D=39.5+/-17.8 nmol/l; P<0.001) while no difference was found for SHBG or LH. Eleven (for E(2), 19 of 40, for 25(OH)D, 20 of 40) out of 40 patients had serum testosterone levels 2 s.d. below the mean of normal. Men with CF showed a relative shift from FBM to LBM and a different body fat distribution compared with healthy controls (P<0.01). Testosterone was not correlated with weight, total or regional LBM or FBM, but significantly with BMD (r=0.32; P<0.05) independently from body height and 25(OH)D levels. E(2) was correlated with regional and total FBM (r=0.48; P<0.05). In a multiple regression analysis of the joint effect of testosterone and body components on E(2), a testosterone independent effect was found for FBM. CONCLUSIONS: CF patients with stable disease have moderately reduced serum testosterone levels. This might already imply detrimental effects on bone. The change in LBM of patients appears to have no direct association with sex hormone levels while low FBM might cause reduced net conversion of serum testosterone to E(2) with possible effects on FBM distribution. PMID- 12720540 TI - Serum concentrations of activin and follistatin are elevated and run in parallel in patients with septicemia. AB - OBJECTIVE: Activin is a growth and differentiation factor of many cell types, and has recently been implicated in inflammatory processes. Clinical data linking activin and its binding protein, follistatin (FS), are lacking. We measured serum levels of activin and FS in patients diagnosed with septicemia. PATIENTS AND MEASUREMENTS: Eight male and seven female patients of different ages, various forms of septicemia and different clinical outcome were investigated and compared with age- and sex-matched healthy controls. Serum concentrations of FS, activin, C-reactive protein (CRP) and blood leukocyte counts were determined during septicemia. RESULTS: The median of the maximum activin concentrations of septicemic patients was 3.9-fold higher than in age- and sex-matched healthy control subjects (P<0.01); the median of the maximum FS concentrations was 2.6 fold higher (P<0.01). The highest increase of activin in septicemic patients was approximately 15.8-fold, whereas FS increased by up to 13.2-fold above normal. FS, activin and CRP serum levels generally paralleled each other, but were not correlated with leukocyte counts or clinical outcome. CONCLUSIONS: Circulatory concentrations of activin and FS are elevated in patients diagnosed with septicemia, consistent with potential roles in the systemic inflammatory response. PMID- 12720541 TI - Autoantibodies to steroidogenic enzymes in patients with premature ovarian failure with and without Addison's disease. AB - DESIGN: Adrenal cortex autoantibodies (ACA), steroid-producing cell autoantibodies (StCA) and autoantibodies (Abs) to steroidogenic enzymes in three groups of patients with premature ovarian failure (POF), 15 with autoimmune Addison's disease (AD), 26 with non-adrenal autoimmune diseases and 31 with isolated POF, have been assessed. METHODS: ACA and StCA were measured using an immunofluorescence technique. Abs to 21-hydroxylase (21-OH), to 17alpha hydroxylase (17alpha-OH) and to cytochrome P450 side-chain cleavage (P450scc) were measured using an immunoprecipitation assay. RESULTS: Seventy-three percent of patients with POF and AD were positive for StCA, 93% for 17alpha-OH and/or P450scc Abs, 93% for ACA and 100% for 21-OH Abs. Among patients with POF and non adrenal autoimmune diseases, 8% were positive for StCA, 12% for 17alpha-OH and/or P450scc Abs, and 8% and 12% for ACA and 21-OH Abs respectively. StCA, 17alpha-OH and/or P450scc Abs were all found in 10% of patients with isolated POF, and 13% had ACA and 21-OH Abs. All StCA-, 17alpha-OH- and/or P450scc Abs-positive patients were also positive for ACA and 21-OH Abs. Two patients with isolated POF who were ACA and 21-OH Ab positive developed AD 3 and 5 Years after the onset of POF. CONCLUSION: This study has shown that, when POF is associated with AD, StCA, 17alpha-OH and/or P450scc Abs are present in the majority of patients, while in the other two groups these Abs are detectable in a much lower proportion of patients. Measurement of ACA/21-OH Abs in some patients with POF may be important in identifying patients at risk of developing overt AD. PMID- 12720542 TI - Progesterone effects during sequential hormone replacement therapy. AB - OBJECTIVE: The aim was to investigate the effect on mood and the physical symptoms of two dosages of natural progesterone and a placebo in postmenopausal women with and without a history of premenstrual syndrome (PMS). DESIGN: A randomized, placebo-controlled, double-blind, crossover study was performed. METHOD: Postmenopausal women (n=36) with climacteric symptoms were recruited. They received 2 mg estradiol continuously during three 28-day cycles. Vaginal progesterone suppositories with 800 mg/day, 400 mg/day, or placebo were added sequentially for 14 days per cycle. Daily symptom ratings using a validated rating scale were kept. RESULTS: Women without a history of PMS showed cyclicity in both negative mood and physical symptoms while on 400 mg/day progesterone but not on the higher dose or the placebo. Women without a history of PMS had more physical symptoms on progesterone treatment compared with placebo. Women with prior PMS reported no progesterone-induced symptom cyclicity. CONCLUSION: In women without prior PMS natural progesterone caused negative mood effects similar to those induced by synthetic progestogens. PMID- 12720543 TI - Expression of cAMP response element-binding protein and sodium iodide symporter in benign non-functioning and malignant thyroid tumours. AB - OBJECTIVE: Reduced expression or defective targeting of the sodium/iodide symporter (NIS) to the cell membrane in thyroid tumours has been reported. The expression of the NIS gene is up-regulated by TSH through the cAMP pathway and the characterization of the promoter region of the rat NIS gene revealed the existence of a degenerate cAMP response element (CRE) sequence. The cAMP dependent transcription factor cAMP response element-binding protein (CREB) binds to CRE acting, upon phosphorylation, as a transcriptional activator. In this study we evaluated the expression of CREB and NIS gene in thyroid non-functioning adenomas (n=18) and carcinomas (n=20), as well as in the corresponding normal tissue. METHODS: The levels of CREB and NIS mRNA were determined by quantitative real-time RT-PCR, whereas CREB protein (total and phosphorylated) was analyzed by Western blot. RESULTS: The levels of CREB mRNA in thyroid carcinomas, but not in adenomas, were significantly lower than in the corresponding normal tissue (4.63+/-0.89 vs 9.51+/-2.01 pg/microg total RNA, means+/-s.e., P=0.025). CREB protein levels, which were determined in a subset of samples, were in quite good agreement with mRNA data. NIS mRNA levels did not differ in adenomas or carcinomas, compared with the corresponding normal tissue and no significant relationship with the levels of CREB mRNA was observed. CONCLUSIONS: Our results have indicated for the first time that reduced levels of CREB expression are a feature of thyroid carcinomas, and confirm that different factors are likely to modulate NIS expression. PMID- 12720545 TI - Biochemical and genetic characterization of a murine class Kappa glutathione S transferase. AB - The class Kappa family of glutathione S-transferases (GSTs) currently comprises a single rat subunit (rGSTK1), originally isolated from the matrix of liver mitochondria [Harris, Meyer, Coles and Ketterer (1991) Biochem. J. 278, 137-141; Pemble, Wardle and Taylor (1996) Biochem. J. 319, 749-754]. In the present study, an expressed sequence tag (EST) clone has been identified which encodes a mouse class Kappa GST (designated mGSTK1). The EST clone contains an open reading frame of 678 bp, encoding a protein composed of 226 amino acid residues with 86% sequence identity with the rGSTK1 polypeptide. The mGSTK1 and rGSTK1 proteins have been heterologously expressed in Escherichia coli and purified by affinity chromatography. Both mouse and rat transferases were found to exhibit GSH conjugating and GSH-peroxidase activities towards model substrates. Analysis of expression levels in a range of mouse and rat tissues revealed that the mRNA encoding these enzymes is expressed predominantly in heart, kidney, liver and skeletal muscle. Although other soluble GST isoenzymes are believed to reside primarily within the cytosol, subcellular fractionation of mouse liver demonstrates that this novel murine class Kappa GST is associated with mitochondrial fractions. Through the use of bioinformatics, the genes encoding the mouse and rat class Kappa GSTs have been identified. Both genes comprise eight exons, the protein coding region of which spans approx. 4.3 kb and 4.1 kb of DNA for mGSTK1 and rGSTK1 respectively. This conservation in primary structure, catalytic properties, tissue-specific expression, subcellular localization and gene structure between mouse and rat class Kappa GSTs indicates that they perform similar physiological functions. Furthermore, the association of these enzymes with mitochondrial fractions is consistent with them performing a specific conserved biological role within this organelle. PMID- 12720544 TI - Ca(2+)-independent protein kinase C activity is required for alpha1-adrenergic receptor-mediated regulation of ribosomal protein S6 kinases in adult cardiomyocytes. AB - The alpha(1)-adrenergic agonist, phenylephrine (PE), exerts hypertrophic effects in the myocardium and activates protein synthesis. Both Ca(2+)-dependent protein kinase C (PKC, PKCalpha) and Ca(2+)-independent PKC isoforms (PKCdelta and epsilon ) are detectably expressed in adult rat cardiomyocytes. Stimulation of the alpha(1)-adrenergic receptor by PE results in activation of Ca(2+) independent PKCs, as demonstrated by translocation of the delta and epsilon isoenzymes from cytosol to membrane fractions. PE also induces activation of p70 ribosomal protein S6 kinases (S6K1 and 2) in adult cardiomyocytes. We have studied the role of Ca(2+)-independent PKCs in the regulation of S6K activity by PE. Activation of S6K1/2 by PE was blocked by the broad-spectrum PKC inhibitor bisindolylmaleimide (BIM) I, whereas Go6976, a compound that only inhibits Ca(2+) dependent PKCs, did not inhibit S6K activation. Rottlerin, which selectively inhibits PKCdelta, also prevented PE-induced S6K activation. The isoform-specific PKC inhibitors had similar effects on the phosphorylation of eukaryotic initiation factor 4E (eIF4E)-binding protein 1, a translation repressor that, like the S6Ks, lies downstream of the mammalian target of rapamycin (mTOR). Infection of cells with adenoviruses encoding dominant-negative PKCdelta or epsilon inhibited the activation of extracellular-signal-regulated kinase (ERK) by PE, and also inhibited the activation and/or phosphorylation of S6Ks 1 and 2. The PE-induced activation of protein synthesis was abolished by BIM I and markedly attenuated by rottlerin. Our data thus suggest that Ca(2+)-independent PKC isoforms play an important role in coupling the alpha(1)-adrenergic receptor to mTOR signalling and protein synthesis in adult cardiomyocytes. PMID- 12720546 TI - A novel cytochrome c peroxidase from Neisseria gonorrhoeae: a lipoprotein from a Gram-negative bacterium. AB - A cytochrome c peroxidase (CCP) produced by Neisseria gonorrhoeae has been shown to have novel characteristics by investigating its location, expression and role in Neisseria gonorrhoeae and by expression in Escherichia coli. Analysis of the N terminus of CCP indicated that it is a lipoprotein with a signal peptide for cleavage by signal peptidase II. Expression of the gonococcal CCP in E. coli revealed that it is first synthesized as a pro-apo-cytochrome that is translocated across the cytoplasmic membrane. The signal peptide is cleaved and haem is attached in the periplasm. The gonococcal CCP was associated with the membrane of both E. coli and N. gonorrhoeae. The expression of a MalE-CCP fusion protein has allowed characterization of CCP in vitro. Evidence is presented that CCP protects gonococci from hydrogen peroxide, presumably in the periplasmic compartment of the cell. The expression of CCP is dependent on the transcription factor FNR, but is repressed by nitrite, indicating that it could be most important in the stationary-phase response. These data support the hypothesis that the gonococcal lipoprotein CCP is anchored to the membrane in the periplasm, where it might be responsible for the reduction of hydrogen peroxide. Other putative CCP lipoproteins have been identified, representing a new subclass of bacterial CCP proteins. PMID- 12720547 TI - Evidence that TRPC1 (transient receptor potential canonical 1) forms a Ca(2+) permeable channel linked to the regulation of cell volume in liver cells obtained using small interfering RNA targeted against TRPC1. AB - The TRPC1 (transient receptor potential canonical 1) protein, which is thought to encode a non-selective cation channel activated by store depletion and/or an intracellular messenger, is expressed in a number of non-excitable cells. However, the physiological functions of TRPC1 are not well understood. The aim of these studies was to investigate the function of TRPC1 in liver cells using small interfering RNA (siRNA) to ablate the TRPC1 protein. Treatment of H4-IIE liver cells with siRNA targeted against TRPC1 caused an approx. 50% decrease in expression of the human TRPC1 protein in cells transfected with cDNA encoding human TRPC1, and a 50% decrease in expression of the endogenous TRPC1 protein (assessed by Western blot and immunofluorescence). The decrease in endogenous TRPC1 protein in cells transfected with TRPC1 siRNA was associated with a greater increase in cell volume (compared with the increase observed in control cells) immediately after cells were placed in a hypotonic medium, and an enhanced regulatory cell volume decrease after exposure to hypotonic medium. Treatment with siRNA targeted against TRPC1 also led to a 25% inhibition of thapsigargin stimulated Ca(2+) inflow, a 40% inhibition of ATP and maitotoxin-stimulated Ca(2+) inflow, and a 50% inhibition of maitotoxin-stimulated Mn(2+) inflow. The idea that, in liver cells, TRPC1 encodes a non-selective cation channel involved directly or indirectly in the regulation of cell volume is consistent with the results obtained. PMID- 12720548 TI - p300/CBP-associated factor (P/CAF) interacts with nuclear respiratory factor-1 to regulate the UDP-N-acetyl-alpha-d-galactosamine: polypeptide N acetylgalactosaminyltransferase-3 gene. AB - We demonstrated recently that expression of the UDP- N -acetyl-alpha-D galactosamine: polypeptide N -acetylgalactosaminyltrans-ferase-3 (GalNAc-T3) gene is restricted to epithelial glands [Nomoto, Izumi, Ise, Kato, Takano, Nagatani, Shibao, Ohta, Imamura, Kuwano, Matsuo, Yamada, Itoh and Kohno (1999) Cancer Res. 59, 6214-6222]. In the present study, we show that sodium butyrate treatment of human breast cancer MCF-7 cells transcriptionally activates the GalNAc-T3 gene. Transient transfection of plasmids containing a reporter gene under the control of GalNAc-T3 indicated that several transcriptional elements are involved in response to sodium butyrate, with the nuclear respiratory factor-1 (NRF-1) binding motif located between -88 and -77nt being the most important. Incubation of a labelled probe encompassing the NRF-1-binding motif with a nuclear extract of sodium butyrate-treated MCF-7 cells yielded a higher level of specific DNA protein complex versus controls. Flag-tagged NRF-1 expressed in MCF-7 cells can bind to the NRF-1-binding motif of the GalNAc-T3 promoter. Nuclear content of NRF 1 remained constant in MCF-7 cells treated with or without sodium butyrate. Moreover, NRF-1 interacts with and is acetylated by p300/CBP-associated factor (P/CAF). Acetylation of NRF-1 enhances DNA binding. Co-transfection of the GalNAc T3 reporter plasmid with either NRF-1 or P/CAF expression plasmid resulted in the activation of the GalNAc-T3 promoter. These results indicate a correlation between acetylation of NRF-1 by P/CAF and the butyrate-induced expression of the GalNAc-T3 gene. Additionally, induced expression of P/CAF may be a component of the adenocarcinoma differentiation process. PMID- 12720549 TI - DNA microarray data and contextual analysis of correlation graphs. AB - BACKGROUND: DNA microarrays are used to produce large sets of expression measurements from which specific biological information is sought. Their analysis requires efficient and reliable algorithms for dimensional reduction, classification and annotation. RESULTS: We study networks of co-expressed genes obtained from DNA microarray experiments. The mathematical concept of curvature on graphs is used to group genes or samples into clusters to which relevant gene or sample annotations are automatically assigned. Application to publicly available yeast and human lymphoma data demonstrates the reliability of the method in spite of its simplicity, especially with respect to the small number of parameters involved. CONCLUSIONS: We provide a method for automatically determining relevant gene clusters among the many genes monitored with microarrays. The automatic annotations and the graphical interface improve the readability of the data. A C++ implementation, called Trixy, is available from http://tagc.univ-mrs.fr/bioinformatics/trixy.html. PMID- 12720550 TI - Pro/con clinical debate: pulmonary artery catheters increase the morbidity and mortality of intensive care unit patients. PMID- 12720551 TI - Low tidal volume, high respiratory rate and auto-PEEP: the importance of the basics. PMID- 12720552 TI - Selective decontamination of the digestive tract reduces mortality in critically ill patients. PMID- 12720553 TI - Recently published papers: changing practices in the modern intensive care unit. PMID- 12720554 TI - Why measure cardiac output? PMID- 12720555 TI - Coagulation cascade in sepsis: getting from bench to bedside? PMID- 12720556 TI - Red man syndrome. PMID- 12720557 TI - ICU cornerstone: 'triggering effort'. PMID- 12720558 TI - Science review: role of coagulation protease cascades in sepsis. AB - Cellular signaling by proteases of the blood coagulation cascade through members of the protease-activated receptor (PAR) family can profoundly impact on the inflammatory balance in sepsis. The coagulation initiation reaction on tissue factor expressing cells signals through PAR1 and PAR2, leading to enhanced inflammation. The anticoagulant protein C pathway has potent anti-inflammatory effects, and activated protein C signals through PAR1 upon binding to the endothelial protein C receptor. Activation of the coagulation cascade and the downstream endothelial cell localized anticoagulant pathway thus have opposing effects on systemic inflammation. This dichotomy is of relevance for the interpretation of preclinical and clinical data that document nonuniform responses to anticoagulant strategies in sepsis therapy. PMID- 12720560 TI - Clinical review: extracorporeal blood purification in severe sepsis. AB - Sepsis and septic shock are the leading causes of acute renal failure, multiple organ system dysfunction, and death in the intensive care unit. The pathogenesis of sepsis is complex and comprises a mosaic of interconnected pathways. Several attempts to improve patient outcomes by targeting specific components of this network have been unsuccessful. For these reasons, the ideal immunomodulating strategy would be one that restores immunologic stability rather than blindly inhibiting or stimulating one or another component of this complex network. Hence, the recent focus of immunomodulatory therapy in sepsis has shifted to nonspecific methods of influencing the entire inflammatory response without suppressing it. Here, we discuss the various modalities of extracorporeal blood purification, the existing evidence, and future prospects. PMID- 12720559 TI - Bench-to-bedside review: endothelial cell dysfunction in severe sepsis: a role in organ dysfunction? AB - During the past decade a unifying hypothesis has been developed to explain the vascular changes that occur in septic shock on the basis of the effect of inflammatory mediators on the vascular endothelium. The vascular endothelium plays a central role in the control of microvascular flow, and it has been proposed that widespread vascular endothelial activation, dysfunction and eventually injury occurs in septic shock, ultimately resulting in multiorgan failure. This has been characterized in various models of experimental septic shock. Now, direct and indirect evidence for endothelial cell alteration in humans during septic shock is emerging. The present review details recently published literature on this rapidly evolving topic. PMID- 12720561 TI - Poisoned patients as potential organ donors: postal survey of transplant centres and intensive care units. AB - BACKGROUND: The number of patients awaiting allograft transplantation in the UK exceeds the number of organs offered for transplantation each year. Most organ donors tend to be young, fit and healthy individuals who die because of trauma or sudden cardiac arrest. Patients who die from drug and poison intoxication tend to have similar characteristics but are less frequently offered as potential organ donors. METHODS: A postal questionnaire survey of all transplantation centres and an equal number of intensive care units in the UK was undertaken. The use of kidney, heart, lung, liver and pancreas transplants from poisoned patients following deliberate methanol ingestion, cardiac arrest presumed secondary to cocaine overdose, accidental domestic carbon monoxide inhalation and industrial cyanide exposure were used as case scenarios. RESULTS: Response rates were 70% for transplantation centres and 50% for intensive care unit directors. Over 80% of organs would be offered or discussed with transplant coordinators by intensive care unit directors. Transplantation physicians/surgeons would consider transplanting organs in up to 100% of case scenarios, depending on the organ and poisoning or intoxication involved. DISCUSSION: The postal survey presented here shows that most transplantation physicians and surgeons and intensive care unit directors would consider those who die following acute drug intoxication and poisoning as potential organ donors. The previously reported literature shows in general that transplanted organs from poisoned patients have good long-term survival, although the number of reports is small. Poisoned patients are another pool of organ donors who at present are probably underused by transplantation services. PMID- 12720562 TI - Safety assessment of drotrecogin alfa (activated) in the treatment of adult patients with severe sepsis. AB - INTRODUCTION: Drotrecogin alfa (activated; recombinant activated protein C) was shown to reduce 28-day all-cause mortality in patients with severe sepsis and to have an acceptable safety profile in 1690 patients studied in the F1K-MC-EVAD (PROWESS) trial. We analyzed all available data on the safety of treatment with drotrecogin alfa (activated) in 2786 adult patients with severe sepsis enrolled in all phase 2 and 3 clinical trials, and in an estimated 3991 patients receiving the drug in commercial use. PATIENTS AND METHOD: Mortality and safety analyses were performed on all available data from adult severe sepsis patients enrolled in seven clinical trials as of 12 April 2002. Trial-specific safety data and spontaneously reported serious adverse events from commercial use were extracted from a pharmacovigilance database. RESULTS: The 28-day mortality rate for all adult patients who received active treatment in all clinical trials was 25.3% (704/2786). Serious bleeding events during the infusion period and 28-day study period occurred in 2.8% (79/2786) and 5.3% (148/2786) of patients, respectively. Of bleeding events during the infusion period, 43% (34/79) were procedure related. Fatal serious bleeding events during the infusion period occurred in 0.4% (12/2786) of cases. Intracranial hemorrhage (ICH) events during the infusion period and 28-day study period occurred in 0.6% (16/2786) and 1.1% (32/2786) of patients, respectively. Ten out of the 16 ICH events occurring during the study drug infusion period were associated with severe thrombocytopenia ( 45%; and group B, with preinduction RVEF < 40%. RESULTS: In group A (n = 10), at time point 3 compared with time point 2, the heart rate significantly decreased (from 75 +/- 10 to 66 +/- 7 beats/min). The right ventricular end diastolic volume index (from 105 +/- 17 to 133 +/- 29 ml/m2), the right ventricular end systolic volume index (from 61 +/- 13 to 77 +/- 24 ml/m2), the systolic systemic arterial/right ventricular pressure gradient (from 93 +/- 24 to 113 +/- 22 mmHg) and the diastolic systemic arterial/right ventricular pressure gradient (from 58 +/- 11 to 66 +/- 12 mmHg) significantly increased. Also in group A, the cardiac index did not significantly increase (from 3.28 +/- 0.6 to 3.62 +/- 0.6 l/min/m2), the RVEF was unchanged, and the right ventricular end diastolic volume/pressure ratio (RVED V/P) did not significantly decrease (from 48 +/- 26 to 37 +/- 13 ml/mmHg). In group B (n = 6) at the same time, the heart rate (from 72 +/- 15 to 66 +/- 12 beats/min), the right ventricular end diastolic volume index (from 171 +/- 50 to 142 +/- 32 ml/m2) and the RVED V/P (from 71 +/- 24 to 39 +/- 7 ml/mmHg) significantly decreased. The cardiac index and the diastolic systemic arterial/right ventricular pressure gradient were unchanged in group B, while the RVEF and the systolic systemic arterial/right ventricular pressure gradient did not significantly increase, and the right ventricular end-systolic volume index did not significantly decrease. All results are expressed as mean +/- standard deviation. CONCLUSIONS: We conclude that passive leg elevation caused a worse condition in the right ventricle of group B because, with stable values of cardiac index, of systolic systemic arterial/right ventricular pressure gradient and of diastolic systemic arterial/right ventricular pressure gradient (which supply oxygen), the RVED V/P (to which oxygen consumption is inversely related) markedly decreased. This is as opposed to group A, where the cardiac index, the systolic systemic arterial/right ventricular pressure gradient and the diastolic systemic arterial/right ventricular pressure gradient increased, and the RVED V/P slightly decreased. Passive leg elevation must therefore be performed cautiously in coronary patients with a reduced RVEF. PMID- 12720564 TI - Comparison of bedside measurement of cardiac output with the thermodilution method and the Fick method in mechanically ventilated patients. AB - INTRODUCTION: Bedside cardiac output determination is a common preoccupation in the critically ill. All available methods have drawbacks. We wished to re-examine the agreement between cardiac output determined using the thermodilution method (QTTHERM) and cardiac output determined using the metabolic (Fick) method (QTFICK) in patients with extremely severe states, all the more so in the context of changing practices in the management of patients. Indeed, the interchangeability of the methods is a clinically relevant question; for instance, in view of the debate about the risk-benefit balance of right heart catheterization. PATIENTS AND METHODS: Eighteen mechanically ventilated passive patients with a right heart catheter in place were studied (six women, 12 men; age, 39-84 years; simplified acute physiology scoreII, 39-111). QTTHERM was obtained using a standard procedure. QTFICK was measured from oxygen consumption, carbon dioxide production, and arterial and mixed venous oxygen contents. Forty nine steady-state pairs of measurements were performed. The data were normalized for repeated measurements, and were tested for correlation and agreement. RESULTS: The QTFICK value was 5.2 +/- 2.0 l/min whereas that of QTTHERM was 5.8 +/- 1.9 l/min (R = 0.840, P < 0.0001; mean difference, -0.7 l/min; lower limit of agreement, -2.8 l/min; upper limit of agreement, 1.5 l/min). The agreement was excellent between the two techniques at QTTHERM values <5 l/min but became too loose for clinical interchangeability above this value. Tricuspid regurgitation did not influence the results. DISCUSSION AND CONCLUSIONS: No gold standard is established to measure cardiac output in critically ill patients. The thermodilution method has known limitations that can lead to inaccuracies. The metabolic method also has potential pitfalls in this context, particularly if there is increased oxygen consumption within the lungs. The concordance between the two methods for low cardiac output values suggests that they can both be relied upon for clinical decision making in this context. Conversely, a high cardiac output value is more difficult to rely on in absolute terms. PMID- 12720565 TI - Training in data definitions improves quality of intensive care data. AB - BACKGROUND: Our aim was to assess the contribution of training in data definitions and data extraction guidelines to improving quality of data for use in intensive care scoring systems such as the Acute Physiology and Chronic Health Evaluation (APACHE) II and Simplified Acute Physiology Score (SAPS) II in the Dutch National Intensive Care Evaluation (NICE) registry. METHODS: Before and after attending a central training programme, a training group of 31 intensive care physicians from Dutch hospitals who were newly participating in the NICE registry extracted data from three sample patient records. The 5-hour training programme provided participants with guidelines for data extraction and strict data definitions. A control group of 10 intensive care physicians, who were trained according the to train-the-trainer principle at least 6 months before the study, extracted the data twice, without specific training in between. RESULTS: In the training group the mean percentage of accurate data increased significantly after training for all NICE variables (+7%, 95% confidence interval 5%-10%), for APACHE II variables (+6%, 95% confidence interval 4%-9%) and for SAPS II variables (+4%, 95% confidence interval 1%-6%). The percentage data error due to nonadherence to data definitions decreased by 3.5% after training. Deviations from 'gold standard' SAPS II scores and predicted mortalities decreased significantly after training. Data accuracy in the control group did not change between the two data extractions and was equal to post-training data accuracy in the training group. CONCLUSION: Training in data definitions and data extraction guidelines is an effective way to improve quality of intensive care scoring data. PMID- 12720566 TI - Protein-losing enteropathy in patients with Fontan circulation: is it triggered by infection? AB - INTRODUCTION: Protein-losing enteropathy (PLE) is a recognised complication of the Fontan circulation. Its pathogenesis is not fully understood, however, and it is unclear why its onset occurs months or even years after Fontan surgery. PATIENTS: We report a 4.5-year-old girl with Fontan circulation who developed PLE almost 1 year after surgery. At the time of onset the patient had rotavirus enteritis and streptococcal tonsillitis. We have reviewed the records of seven other patients with longstanding PLE. In six of these patients we identified infections at the onset of symptoms. None of our patients had evidence of opportunistic infection. DISCUSSION AND CONCLUSION: The immune system of patients with PLE is compromised, but reports on recurrent opportunistic infections are rare. The present observations suggest that infection and inflammation may be associated with the onset of PLE. The mechanism of how infection may trigger PLE warrants further investigation. PMID- 12720568 TI - Positive end-expiratory pressure or no positive end-expiratory pressure: is that the question to be asked? PMID- 12720567 TI - Prophylactic positive end-expiratory pressure: are good intentions enough? PMID- 12720569 TI - The 32nd Annual Congress of the Society of Critical Care Medicine, 28 January - 2 February 2003, San Antonio, USA. PMID- 12720570 TI - Reducing mortality in sepsis: new directions. AB - Considerable progress has been made in the past few years in the development of therapeutic interventions that can reduce mortality in sepsis. However, encouraging physicians to put the results of new studies into practice is not always simple. A roundtable was thus convened to provide guidance for clinicians on the integration and implementation of new interventions into the intensive care unit (ICU). Five topics were selected that have been shown in randomized, controlled trials to reduce mortality: limiting the tidal volume in acute lung injury or acute respiratory distress syndrome, early goal-directed therapy, use of drotrecogin alfa (activated), use of moderate doses of steroids, and tight control of blood sugar. One of the principal investigators for each study was invited to participate in the roundtable. The discussions and questions that followed the presentation of data by each panel member enabled a consensus recommendation to be derived regarding when each intervention should be used. Each new intervention has a place in the management of patients with sepsis. Furthermore, and importantly, the therapies are not mutually exclusive; many patients will need a combination of several approaches--an "ICU package". The present article provides guidelines from experts in the field on optimal patient selection and timing for each intervention, and provides advice on how to integrate new therapies into ICU practice, including protocol development, so that mortality rates from this disease process can be reduced. PMID- 12720571 TI - Toward a better understanding of the comparatively high prostate cancer incidence rates in Utah. AB - BACKGROUND: This study assesses whether comparatively high prostate cancer incidence rates among white men in Utah represent higher rates among members of the Church of Jesus Christ of Latter-day Saints (LDS or Mormons), who comprise about 70% of the state's male population, and considers the potential influence screening has on these rates. METHODS: Analyses are based on 14,693 histologically confirmed invasive prostate cancer cases among men aged 50 years and older identified through the Utah Cancer Registry between 1985 and 1999. Cancer records were linked to LDS Church membership records to determine LDS status. Poisson regression was used to derive rate ratios of LDS to nonLDS prostate cancer incidence, adjusted for age, disease stage, calendar time, and incidental detection. RESULTS: LDS men had a 31% (95% confidence interval, 26% 36%) higher incidence rate of prostate cancer than nonLDS men during the study period. Rates were consistently higher among LDS men over time (118% in 1985-88, 20% in 1989-92, 15% in 1993-1996, and 13% in 1997-99); age (13% in ages 50-59, 48% in ages 60-69, 28% in ages 70-79, and 16% in ages 80 and older); and stage (36% in local/regional and 17% in unstaged). An age- and stage-shift was observed for both LDS and nonLDS men, although more pronounced among LDS men. CONCLUSIONS: Comparatively high prostate cancer incidence rates among LDS men in Utah are explained, at least in part, by more aggressive screening among these men. PMID- 12720572 TI - Anticonvulsant, sedative and muscle relaxant effects of carbenoxolone in mice. AB - BACKGROUND: Carbenoxolone, as an antiulcer medicine, has some pharmacological properties such as: the inhibition of gap junctional (GJ) intercellular communication. In vitro studies have shown, carbenoxolone to abolish the generation of full or partial ectopic spike generation, by 4-aminopyridine, as well as spontaneous epileptiform activity in CA3 or CA1 regions of the rat hippocampal slices via closing GJ channels. Thus, we considered the possible anticonvulsant effects of carbenoxolone in animal seizure models. RESULTS: ED50 values of diazepam and carbenoxolone in the pentylenetetrazole model were 1.13 mg/kg and 283.3 mg/kg, respectively. In this model, carbenoxolone in doses of 200 and 300 mg/kg prolonged the onset time of seizure and decreased the duration of seizures. In the maximal electroshock model, carbenoxolone in a dose of 400 mg/kg decreased the duration of seizure producing protection against seizure but failing to protect against mortality in comparison with diazepam. In the potentiation of pentobarbitone sleep test, carbenoxolone significantly increased sleeping time and decreased latency in doses of 100, 200 and 300 mg/kg in mice dose dependently. In the traction test, carbenoxolone (400 mg/kg) showed muscle relaxant activity and in the accelerated rotarod test, carbenoxolone in doses of 200 and 300 mg/kg showed a decline in motor coordination. CONCLUSION: It can be concluded that carbenoxolone possesses anticonvulsant, muscle relaxant and hypnotic effects, which could contribute to the control of petit mal and grand mal seizures. PMID- 12720573 TI - A general cloning system to selectively isolate any eukaryotic or prokaryotic genomic region in yeast. AB - BACKGROUND: Transformation-associated recombination (TAR) cloning in yeast is a unique method for selective isolation of large chromosomal fragments or entire genes from complex genomes. The technique involves homologous recombination, during yeast spheroplast transformation, between genomic DNA and a TAR vector that has short (approximately 60 bp) 5' and 3' gene targeting sequences (hooks). RESULT: TAR cloning requires that the cloned DNA fragment carry at least one autonomously replicating sequence (ARS) that can function as the origin of replication in yeast, which prevents wide application of the method. In this paper, we describe a novel TAR cloning system that allows isolation of genomic regions lacking yeast ARS-like sequences. ARS is inserted into the TAR vector along with URA3 as a counter-selectable marker. The hooks are placed between the TATA box and the transcription initiation site of URA3. Insertion of any sequence between hooks results in inactivation of URA3 expression. That inactivation confers resistance to 5-fluoroorotic acid, allowing selection of TAR cloning events against background vector recircularization events. CONCLUSION: The new system greatly expands the area of application of TAR cloning by allowing isolation of any chromosomal region from eukaryotic and prokaryotic genomes regardless of the presence of autonomously replicating sequences. PMID- 12720574 TI - Partial duplication of the APBA2 gene in chromosome 15q13 corresponds to duplicon structures. AB - BACKGROUND: Chromosomal abnormalities affecting human chromosome 15q11-q13 underlie multiple genomic disorders caused by deletion, duplication and triplication of intervals in this region. These events are mediated by highly homologous segments of DNA, or duplicons, that facilitate mispairing and unequal cross-over in meiosis. The gene encoding an amyloid precursor protein-binding protein (APBA2) was previously mapped to the distal portion of the interval commonly deleted in Prader-Willi and Angelman syndromes and duplicated in cases of autism. RESULTS: We show that this gene actually maps to a more telomeric location and is partially duplicated within the broader region. Two highly homologous copies of an interval containing a large 5' exon and downstream sequence are located approximately 5 Mb distal to the intact locus. The duplicated copies, containing the first coding exon of APBA2, can be distinguished by single nucleotide sequence differences and are transcriptionally inactive. Adjacent to APBA2 maps a gene termed KIAA0574. The protein encoded by this gene is weakly homologous to a protein termed X123 that in turn maps adjacent to APBA1 on 9q21.12; APBA1 is highly homologous to APBA2 in the C terminal region and is distinguished from APBA2 by the N-terminal region encoded by this duplicated exon. CONCLUSION: The duplication of APBA2 sequences in this region adds to a complex picture of different low copy repeats present across this region and elsewhere on the chromosome. PMID- 12720575 TI - Evidence for survival of Pleistocene climatic changes in Northern refugia by the land snail Trochoidea geyeri (Soos 1926) (Helicellinae, Stylommatophora). AB - BACKGROUND: The study of organisms with restricted dispersal abilities and presence in the fossil record is particularly adequate to understand the impact of climate changes on the distribution and genetic structure of species. Trochoidea geyeri (Soos 1926) is a land snail restricted to a patchy, insular distribution in Germany and France. Fossil evidence suggests that current populations of T. geyeri are relicts of a much more widespread distribution during more favourable climatic periods in the Pleistocene. RESULTS: Phylogeographic analysis of the mitochondrial 16S rDNA and nuclear ITS-1 sequence variation was used to infer the history of the remnant populations of T. geyeri. Nested clade analysis for both loci suggested that the origin of the species is in the Provence from where it expanded its range first to Southwest France and subsequently from there to Germany. Estimated divergence times predating the last glacial maximum between 25-17 ka implied that the colonization of the northern part of the current species range occurred during the Pleistocene. CONCLUSION: We conclude that T. geyeri could quite successfully persist in cryptic refugia during major climatic changes in the past, despite of a restricted capacity of individuals to actively avoid unfavourable conditions. PMID- 12720576 TI - Vitamin D in preventive medicine: are we ignoring the evidence? AB - Vitamin D is metabolised by a hepatic 25-hydroxylase into 25-hydroxyvitamin D (25(OH)D) and by a renal 1alpha-hydroxylase into the vitamin D hormone calcitriol. Calcitriol receptors are present in more than thirty different tissues. Apart from the kidney, several tissues also possess the enzyme 1alpha hydroxylase, which is able to use circulating 25(OH)D as a substrate. Serum levels of 25(OH)D are the best indicator to assess vitamin D deficiency, insufficiency, hypovitaminosis, adequacy, and toxicity. European children and young adults often have circulating 25(OH)D levels in the insufficiency range during wintertime. Elderly subjects have mean 25(OH)D levels in the insufficiency range throughout the year. In institutionalized subjects 25(OH)D levels are often in the deficiency range. There is now general agreement that a low vitamin D status is involved in the pathogenesis of osteoporosis. Moreover, vitamin D insufficiency can lead to a disturbed muscle function. Epidemiological data also indicate a low vitamin D status in tuberculosis, rheumatoid arthritis, multiple sclerosis, inflammatory bowel diseases, hypertension, and specific types of cancer. Some intervention trials have demonstrated that supplementation with vitamin D or its metabolites is able: (i) to reduce blood pressure in hypertensive patients; (ii) to improve blood glucose levels in diabetics; (iii) to improve symptoms of rheumatoid arthritis and multiple sclerosis. The oral dose necessary to achieve adequate serum 25(OH)D levels is probably much higher than the current recommendations of 5-15 microg/d. PMID- 12720577 TI - Na+/glucose co-transporter abundance and activity in the small intestine of lambs: enhancement by abomasal infusion of casein. AB - The purpose of the present study was to determine the effect of abomasal casein infusion on glucose uptake and abundance of the Na+/glucose co-transporter (SGLT1) 1 in the ovine small intestine. Lambs (body weight 35 (sem 1.0) kg) were surgically fitted with abomasal infusion catheters and were fed diets containing equal portions of wheat hay and cracked maize. Lambs were infused with either 500 g water/d or with 500 g water containing 35 g casein/d. The infusion period lasted 10 d, after which lambs were killed, exsanguinated and eviscerated. Brush border membrane vesicles (BBMV) were prepared using mucosa from different small intestinal regions. Intake and total tract digestibility of nutrients were similar between treatments and averaged 1134, 1142 and 486 g/d and 67, 70, and 94 % for DM, organic matter and non-structural carbohydrates respectively. Crude protein (Nx6.25) digestibility was 15 % greater in the casein-infused than control lambs. Glucose uptake to BBMV ranged from 101 to 337 pmol/mg protein per s along the small intestine and was greatest in the mid-section of the small intestine. In the mid-jejunum, glucose uptake was greater (P<0.07) in lambs infused with casein and averaged 120 pmol/mg protein per s compared with 68 pmol/mg protein per s in the control group. SGLT1 affinity was similar between treatments and averaged 104 microm in the different segments of the small intestine of lambs. However, lambs infused with casein exhibited similar values along the small intestine and affinity averaged 106 microm, while in the control group a greater affinity (85 microm) was measured in the mid-jejunum. SGLT1 protein abundance was correlated with glucose uptake in the BBMV in the casein treated lambs, but not in the control group. These results suggest that glucose uptake along the small intestine of lambs is influenced by casein or its derivatives in the small intestine via SGLT1 affinity and activity at the brush border membrane, and that SGLT1 activity may be regulated by post-translational events affected by amino acids and peptides. PMID- 12720578 TI - Effect of 4-coumaric and 3,4-dihydroxybenzoic acid on oxidative DNA damage in rat colonic mucosa. AB - The effect of 4-coumaric and 3,4-dihydroxybenzoic (protocatechuic) acid on the basal oxidative DNA damage of rat colonic mucosa in vivo was studied, relative to vitamin E. F344 rats were treated with 4-coumaric or protocatechuic acid mixed in the diet (25 or 50 mg/kg for 2 weeks). It was observed that 4-coumaric acid (50 mg/kg) significantly decreased the basal level of the oxidative damage assessed as 8-OH-2'-deoxyguanosine levels in DNA and by the comet assay. Moreover, it was found that vitamin E (10 mg/kg) had no effect on colonic mucosa oxidation damage, whereas at a higher dose (55 mg/kg) it actually enhanced oxidative stress. The effect of 4-coumaric acid (50 mg/kg) on the expression of some glutathione related enzymes (glutathione-S-transferase (GST)-P, GST-M2, GST-M1, gamma glutamylcysteine synthetase, glutathione peroxidase (GSPX)1 and GSPX4) was also investigated at the level of the colonic mucosa. Only the expression of GST-M2 was significantly induced by 4-coumaric acid, while protocatechuic acid was inactive. The data suggest that 4-coumaric acid acts as an antioxidant in the colonic mucosa in vivo. PMID- 12720579 TI - Interaction between vitamins C and E affects their tissue concentrations, growth, lipid oxidation, and deficiency symptoms in yellow perch (Perca flavescens). AB - We have conducted studies with juvenile yellow perch (Perca flavescens) over a period of 20 weeks to address the question of the interaction between water- and lipid-soluble antioxidant vitamins. Fish (2.25+/-0.14 g) were divided into twelve groups, and triplicate groups were fed one of four casein-based, semi-purified diets formulated to contain low or high vitamin E levels of either 5 or 160 mg/kg without or with vitamin C supplementation (250 mg/kg). Diets were designated as C-E, -C+E, +C-E, or +C+E, respectively. The fish fed the +C+E diet showed significantly higher weight gain, feed intake, and feed efficiency than the groups fed vitamin C-deficient diets. Total ascorbate concentrations of liver were significantly higher in fish fed vitamin C-supplemented diets than in fish fed the vitamin C-deficient diet after 16 and 20 weeks. The liver alpha tocopherol concentrations were increased by supplemental vitamin C in vitamin E deficient dietary groups which indicates a sparing or regenerating effect of vitamin C on vitamin E. Fish fed vitamin C-deficient diets (-C-E and -C+E) exhibited severe deficiency symptoms, such as scoliosis, lens cataracts, anorexia, and haemorrhages. The cumulative mortality was significantly higher in the -C-E groups. The thiobarbituric acid-reactive substances value was significantly higher in blood plasma of fish fed a diet unsupplemented with both vitamins. The findings in the present study with yellow perch support the hypothesis that vitamin C regenerates and/or spares vitamin E in vivo. PMID- 12720580 TI - Fructans in the diet cause alterations of intestinal mucosal architecture, released mucins and mucosa-associated bifidobacteria in gnotobiotic rats. AB - The effects of fructans in the diet on the mucosal morphometry (height of villi, depth of the crypts, number of goblet cells), the thickness of the epithelial mucus layer and the histochemical composition of intestinal mucosubstances in the distal jejunum and the distal colon were investigated by comparing germ-free (GF) rats, rats harbouring Bacteroides vulgatus and Bifidobacterium longum (diassociated (DA) rats), and rats with a human faecal flora (HFA). The rats were fed either a commercial standard diet (ST) or ST + (50 g oligofructose (OF)-long chain inulin (lcIN))/kg. Changes in total bacteria, bifidobacteria and Bacteroides-Prevotella in response to feeding these diets were investigated by fluorescent in situ hybridization with 16S rRNA-targeted probes both in intestinal contents (lumen bacteria) and tissue sections (mucosa-associated bacteria). The OF-lcIN-containing diet resulted in higher villi and deeper crypts in bacteria-associated, but not in GF rats. In DA and HFA rats, the colonic epithelial mucus layer was thicker and the numbers of the goblet cells were greater than in GF rats. These effects were enhanced by the OF-lcIN-containing diet. In both dietary groups, bacterial colonization of GF rats caused an increase in neutral mucins in the distal jejunum and colon. Bacteria-associated rats had more acidic mucins in the colon than GF rats, and the OF-lcIN-containing diet stimulated sulfomucins as the predominant type of acidic mucins, while sialomucins dominated in the ST-fed groups. The number of mucosa-associated bifidobacteria detected in the colon of DA and HFA rats was greater with OF-lcIN than ST (4.9 and 5.4 v. 3.5 and 4.0 log10/mm2 mucosal surface respectively), whereas the number of luminal bifidobacteria was only affected by fructans in DA rats. Bacteroides did not differ between the groups. The stabilisation of the gut mucosal barrier, either by changes in the mucosal architecture itself, in released mucins or by stimulation of mucosal bifidobacteria with fructans, could become an important topic in the treatment and prophylaxis of gastrointestinal disorders and health maintenance. PMID- 12720581 TI - Effect of genistein and daidzein on platelet aggregation and monocyte and endothelial function. AB - There has been much recent interest in the cardiovascular benefits of dietary isoflavones. The aim of the present in vitro studies was to investigate potential anti-thrombogenic and anti-atherogenic effects of the isoflavones genistein and daidzein in platelets, macrophages and endothelial cells. Pre-treatment with either isoflavone inhibited collagen-induced platelet aggregation in a dose dependent manner. In a macrophage cell line (RAW 264.7) activated with interferon gamma plus lipopolysaccharide, both isoflavones were found to inhibit NO production and tumour necrosis factor alpha (TNF-alpha) secretion dose dependently, but they did not affect mRNA levels for inducible nitric oxide synthase and cyclo-oxygenase-2. Both isoflavones also dose-dependently decreased monocyte chemoattractant protein-1 secretion induced by TNF-alpha in human umbilical vein endothelial cells. Compared with daidzein, genistein exerted greater inhibitory effects for all parameters studied. The present data contributes to our knowledge on the molecular mechanisms by which isoflavones may protect against coronary artery disease. Further studies are required to determine whether the effects of isoflavones observed in the current in vitro studies are relevant to the aetiology of coronary artery disease in vivo. PMID- 12720582 TI - Oxidation of essential amino acids by the ovine gastrointestinal tract. AB - It is not known if the ruminant animal gastrointestinal tract (GIT) can oxidise essential amino acids (AA) other than leucine. Therefore, the oxidation of four essential AA (leucine, lysine, methionine and phenylalanine), supplied systemically as labelled 1-13C forms, was monitored across the mesenteric-drained viscera (MDV; small intestine) and portal-drained viscera (PDV; total GIT), as part of a Latin square design, in four wether sheep (35-45 kg) fed at 1.4 x maintenance. Oxidation was assessed primarily by appearance of 13CO2, corrected for sequestration of [13C]bicarbonate. The GIT contributed 25 % (P<0.001) and 10 % (P<0.05) towards whole-body AA oxidation for leucine and methionine respectively. This reduced net appearance across the PDV by 23 and 11 % respectively. The contribution of MDV metabolism to total PDV oxidation was 40 % for leucine and 60 % for methionine. There was no catabolism of systemic lysine or phenylalanine across the GIT. Production and exchange of secondary metabolites (e.g. 4-methyl-2-oxo-pentanoate, homocysteine, 2-aminoadipate) across the GIT was also limited. Less AA appeared across the PDV than MDV (P<0.001), indicative of use by tissues such as the forestomach, large intestine, spleen and pancreas. The PDV: MDV net appearance ratios varied (P<0.001) between AA, e.g. phenylalanine (0.81), lysine (0.71), methionine (0.67), leucine (0.56), histidine (0.71), threonine (0.63) and tryptophan (0.48). These differences probably reflect incomplete re-absorption of endogenous secretions and, together with the varied oxidative losses measured, will alter the pattern of AA net supply to the rest of the animal. PMID- 12720583 TI - Metabolic effects of trans fatty acids on an experimental dietary model. AB - The aim of the present study was to investigate the potential nutritional and metabolic impact of trans (t) fatty acids (FA) on an appropriate experimental dietary model. Since previously reported experimental designs have been matter of concern, we developed a dietary model to compare the effect of t isomers and/or the saturation of FA independently of other variables. Wistar rats were fed diets containing identical amounts of nutrients and high levels of dietary fats (200 g/kg) for 30 d. Dietary fat rich in t-FA was compared with fat rich in saturated (s) FA or rich in cis (c) FA, maintaining the same length of C chain of the FA. The fats were obtained through isomerization or hydrogenation of the c-FA present in the control fat. Apparent fat absorption, energy efficiency and triacylglycerol levels in serum and liver were different in rats fed t-FA or s-FA than c-FA. The apparent fat absorption was (%): s-FA 85.7 (sd 3.4)24.9 kg/m2). A greater waist circumference in men and women (age-adjusted by covariance analysis) was observed among Brazil-born participants. In general, only 10 % of the participants reported current practice of sports or other vigorous physical activity. Age-adjusted mean energy intakes and % energy from macronutrients were found to be similar across generations. The age-adjusted mean daily % energy intake from fat were similar across generations: among Japan-born participants, they were 31.5 (95 % CI 30.6, 32.4) % for men and 32.6 (95 % CI 31.7, 33.5) % for women. The respective figures for Brazil-born subjects were 32.1 (95 % CI 31.6, 32.6) % and 33.2 (95 % CI 32.7, 33.5) %. These values are quite different from the usual intakes reported in Japan during the last decades (about 25 %). Taking into account the traditional Japanese diet, a high energy density diet and a sedentary lifestyle may be implicated in the high prevalence of central obesity and metabolic syndrome observed among Japanese Brazilian subjects across gender and generations. PMID- 12720590 TI - Zinc supplementation has no effect on circulating levels of peripheral blood leucocytes and lymphocyte subsets in healthy adult men. AB - As a result of evidence documenting harmful effects of Zn supplementation on immune function and Cu status, thirty-eight men were recruited onto a Zn supplementation trial. The aim was to examine the effects of chronic Zn supplementation on circulating levels of peripheral blood leucocytes and lymphocyte subsets. Subjects (n 19) took 30 mg Zn/d for 14 weeks followed by 3 mg Cu/d for 8 weeks to counteract adverse effects, if any, of Zn supplementation on immune status resulting from lowered Cu status. A control group (n 19) took placebo supplements for the duration of the trial. Dietary intakes of Zn approximated 10 mg/d. Blood samples, taken throughout the trial, were assessed for full blood profiles and flow cytometric analyses of lymphocyte subsets. Putative indices of Cu status were also examined. Results indicate that there was no effect of Zn supplementation on circulating levels of peripheral blood leucocytes or on lymphocyte subsets. Cu status was also unaltered. Independent of supplement, there appeared to be seasonal variations in selected lymphocyte subsets in both placebo and supplemented groups. Alterations in circulating levels of B cells (cluster of differentiation (CD) 19), memory T cells (CD45RO) and expression of the intracellular adhesion molecule-1 (CD54) on T cells were observed. Findings indicated no adverse effects of Zn supplementation on immune status or Cu status and support the US upper level of Zn tolerance of 40 mg/d. The seasonal variations observed in lymphocyte subsets in the group as a whole could have implications for seasonal variability in the incidence of infectious diseases. PMID- 12720592 TI - Energy balance in depleted ambulatory patients with chronic obstructive pulmonary disease: the effect of physical activity and oral nutritional supplementation. AB - Patients with chronic obstructive pulmonary disease (COPD) often suffer from weight loss. The aim of the present study was to gain insight into the energy balance of depleted ambulatory COPD patients, in relation to their habitual level of physical activity and consumption of oral nutritional supplements. Clinically stable and weight-stable patients (n 20; BMI 19.8+/- SD 2.0 kg/m2) were studied 1 and 3 months after rehabilitation or recovery in the clinic and were at random assigned to a control or intervention group with regard to nutritional supplementation. Energy intake was measured with a 7 d food record. Energy expenditure was estimated from a simultaneous 7 d assessment of physical activity with a tri-axial accelerometer for movement registration in combination with measured BMR. Body mass was measured at several time points. The body mass remained stable in both groups after 1 or 3 months and mean energy balances were comparable for both groups. The mean body-mass change between month 1 and 3 was negatively related to the mean physical activity level (r -0.49; P=0.03). Weight change over the 3 months was negatively associated with the physical activity level. These results suggest that knowledge about the individual physical activity level is necessary for the estimation of the energy need of the COPD patient. PMID- 12720593 TI - The effects of exercise and protein-energy supplements on body composition and muscle function in frail elderly individuals: a long-term controlled randomised study. AB - Fighting against inactivity and inadequate nutritional intake are of utmost importance in the elderly. To our knowledge, the few studies which have been performed were conducted for only a short period and the results do not permit formal conclusions to be drawn. We therefore tried to fill this gap in our knowledge by determining whether an intervention combining an acceptable progressive exercise programme and nutritional supplements would be feasible for a long-term period in the very frail elderly, and would bring about concomitant benefits in body composition and muscle power. Accordingly, this exercise and nutritional combination was assessed in the frail elderly in a 9-month randomised trial with a factorial design. Fifty-seven elderly volunteers over 72 years, from sixteen retirement homes in Lyon, France participated in the study. Dietary supplements were compared with placebo, and physical exercise was compared with memory training. Main outcome measures were fat-free mass (FFM) and muscle power. FFM was determined by labelled water, and muscle power was measured by a leg extensor machine. At 9 months, the compliance was 63 % for exercise sessions, and 54 % for nutritional supplements. In patients with dietary supplements, muscle power increased by 57 % at 3 months (P=0.03), and showed only a tendency at 9 months; although FFM increased by 2.7 % at 9 months, the difference was not significant (P=0.10). Exercise did not improve muscle power at 9 months, but improved functional tests (five-time-chair rise, P=0.01). BMI increased with supplements (+3.65 %), but decreased with placebo (-0.5 %) at 9 months (P=0.007). A long-term combined intervention is feasible in frail elderly individuals with a good rate of compliance. Nutritional supplements and exercise may improve muscle function. Despite no significant results on FFM, due to the limited number of volunteers, combined intervention should be suggested to counteract muscle weakness in the frail elderly. PMID- 12720594 TI - Higher erythrocyte 22:6n-3 and 22:5n-6, and lower 22:5n-3 suggest higher Delta-4 desaturation capacity in women of childbearing age. PMID- 12720595 TI - The effect of nutrient profiles of the Dietary Approaches to Stop Hypertension (DASH) diets on blood pressure and bone metabolism and composition in normotensive and hypertensive rats. AB - Hypertension has been associated with abnormalities of Ca and bone metabolism. Consequently, dietary strategies aimed at reducing blood pressure may also benefit bone health; however, this issue has received little attention. Therefore, the objective of the present study was to investigate the effect of two antihypertensive-type diets on blood pressure and bone metabolism and composition in normotensive (Wistar-Kyoto NHsd, WKY) and hypertensive (spontaneously hypertensive NHsd, SHR) rats. Thirty WKY and thirty SHR male rats, 14 weeks old, were separately randomized by weight into three groups of ten rats each. One group from each strain was given a control diet while the other two groups were fed two anti-hypertensive (high fruit and vegetable (F/V) and high fruit and vegetable and low-fat dairy produce (combination)) diets for 8 weeks. SHR rats were significantly (P<0.01) heavier than WKY rats. Blood pressure and femoral length, width, dry weight, ash, Ca, Mg, P and bone mineral mass were significantly (P<0.0001) greater in SHR than WKY rats, but were unaffected by diet, irrespective of strain. While markers of bone formation (serum osteocalcin) and bone resorption (urinary pyridinoline and deoxypyridinoline) were similar in both strains, these markers were significantly (P<0.05) lower (28-31, 16-23, 31 33 % respectively) in the SHR rats fed the combination diet relative to those fed the control and F/V diets. Bone turnover in WKY rats was unaffected by diet. In conclusion, these findings suggest that the combination diet may benefit bone metabolism in hypertensive animals. However, as blood pressure was unaffected by this diet, the mechanism by which it reduced bone turnover requires further investigation. PMID- 12720597 TI - Pharmacotherapy of neuropathic low back pain. AB - Neuropathic low back pain is examined from a structural standpoint, distinguishing processes that start from chronic inflammation and mechanical compromise and cross into the realm of neuropathy with primary neurogenic pathophysiology. The disease of chronic pain is discussed, examining peripheral and central changes in neuroanatomy, neurophysiology, and neuromolecular dynamics. The limitations of inadequate random controlled trials regarding long term pharmacologic interventions are contrasted with excellent work in the basic science of chronic pain. Complex rational pharmacologic strategies for structural pathology, central pain processes, sites of medication action, and differing routes of administration are delineated. PMID- 12720596 TI - Pharmacotherapy of painful diabetic neuropathy. AB - The scope of this review is to describe the epidemiology, physiology, symptomatology, and treatment of diabetic painful neuropathy, which is a common complication of diabetes with significant morbidity. This article focuses on treatment options. Various clinical trials of several classes of medications (eg, antidepressants, anticonvulsants, and topical medications) and alternative treatments (eg, acupuncture, electrostimulation, magnets) are reviewed. Physicians have a large panel of medications that can be used effectively solely or in combination at their disposal. However, a number of these treatments have significant side effects, which are noted, that limit their use. As the understanding of the pathophysiologic mechanisms of diabetic neuropathy improves, new medications are under investigation, which are reviewed in this article. There is great hope that the future may hold treatments that would prevent nerve damage. PMID- 12720598 TI - Complex regional pain syndrome: a review of evidence-supported treatment options. AB - Complex regional pain syndrome consists of pain and other symptoms that are unexpectedly severe or protracted after an injury. In type II complex regional pain syndrome, major nerve injury, often with motor involvement, is the cause; in complex regional pain syndrome I, the culprit is a more occult lesion, often a lesser injury that predominantly affects unmyelinated axons. In florid form, disturbances of vasoregulation (eg, edema) and abnormalities of other innervated tissues (skin, muscle, bone) can appear. Because of these various symptoms and the difficulty in identifying causative lesions, complex regional pain syndrome is difficult to treat or cure. Complex regional pain syndrome has not been systematically investigated; there are few controlled treatment trials for established complex regional pain syndrome. This article reviews the existing studies (even if preliminary) to direct clinicians toward the best options. Treatments for other neuropathic pain syndromes that may be efficacious for complex regional pain syndrome also are discussed. Some common treatments (eg, local anesthetic blockade of sympathetic ganglia) are not supported by the aggregate of published studies and should be used less frequently. Other treatments with encouraging published results (eg, neural stimulators) are not used often enough. We hope to encourage clinicians to rely more on evidence supported treatments for complex regional pain syndrome. PMID- 12720600 TI - Imaging chronic daily headache. AB - New imaging technology allows us to study neurologic disorders that have had no previous structural basis. There have been recent reports on the involvement of nociceptive pathways in daily headache. A systematic review was performed using key words "chronic daily headache" and "imaging." This paper reviews the literature on imaging studies performed on daily headache with emphasis on the new imaging technology. PMID- 12720599 TI - Pharmacotherapy for pain in rheumatologic conditions: the neuropathic component. AB - Nociceptive and neuropathic types of pain occur in rheumatologic conditions. Most clinicians are familiar with the former, but many are not aware of the prevalence of the latter. The literature reports numerous examples of the occurrence of rheumatologic neuropathic pain, but little has been published on its management. In this article, neuropathic and nociceptive pain in rheumatologic conditions are differentiated and treatment recommendations are discussed. Common rheumatologic conditions and their pathophysiology in relation to pain mechanisms also are described. Pharmacotherapeutic recommendations for the treatment of both types of pain in the common rheumatologic conditions are presented. PMID- 12720602 TI - New daily persistent headache. AB - New daily persistent headache was first described by Vanast in 1986 as a benign form of chronic daily headache that improved without therapy. In the headache specialist's office, new daily persistent headache is anything but benign and is thought to be one of the most treatment refractory of all headache conditions. Little is known about this syndrome. It is unique in that the headache begins daily from onset, typically in a patient without a history of headache, and can continue for years without any sign of alleviation despite aggressive treatment. This article discusses the epidemiology, diagnostic criteria, clinical characteristics, and treatment strategies for new daily persistent headache. PMID- 12720601 TI - Migraine genetics. AB - The genetics of migraine is a fascinating and moving research area. Familial hemiplegic migraine, a rare subtype of migraine with a Mendelian pattern of inheritance, is caused by mutations in the chromosome 19 CACNA1A gene in approximately 75% of the families. The finding of mutations in an ionchannel subunit defines migraine as a channelopathy (eg, epilepsy). The genetics of the more frequent variants, migraine with and without aura, is more complex. Several loci have been studied in families and case-control studies, but need to be confirmed. PMID- 12720604 TI - Is there a role for botulinum toxin in the treatment of migraine? AB - In this review, the studies and case reports that are available from reference systems and published congress contributions on the treatment of migraine with botulinum toxin are evaluated. The studies and reports were analyzed with respect to the study design, the efficacy parameters, and the significance of results. One double-blind, placebo-controlled, randomized study with negative (for a 75 U dose of botulinum toxin) and positive (for a 25 U dose of botulinum toxin) evidence of efficacy, one that was a partly positive controlled study (pain intensity, but not attack-frequency improved), and four positive open studies were available. For the acute treatment of migraine with botulinum toxin, only positive case reports were published. As a result of this analysis, there is no sufficient scientific evidence for a treatment recommendation of migraine with botulinum toxin. Further studies are needed for a definite evaluation of subgroups with probable benefit from such a treatment and for the comparison of botulinum toxin with other migraine prophylactic drugs. PMID- 12720605 TI - Malnutrition research: high time to change the menu. PMID- 12720603 TI - Pseudomigraine with lymphocytic pleocytosis. AB - Pseudomigraine with temporary neurologic symptoms and lymphocytic pleocytosis is a self-limited syndrome of unknown origin characterized by headache accompanied by transient neurologic symptoms and cerebrospinal fluid lymphocytosis. Patients with this condition are between 15 and 40 years of age. The syndrome is more frequent in men. The clinical picture encompasses one to 12 episodes of changing variable neurologic deficits accompanied by moderate to severe headache and occasional fever. These headaches are described as predominantly throbbing and bilateral with a variable duration (mean, 19 hours). The average duration of the transient neurologic deficit is 5 hours. Sensory (78% episodes), aphasic (66%), and motor (56%) disturbances are the most common. Migraine-like visual symptoms are relatively rare (18% episodes). Patients are asymptomatic between episodes and after the symptomatic period (duration > 3 months). Lymphocytic pleocytosis (10 to 760 cells mm(3)) and increased cerebrospinal fluid protein are found with negative bacteriologic, viral, fungal, and immunologic studies. Brain computed tomography and magnetic resonance imaging are normal, but an electroencephalogram frequently shows focal slowing over the symptomatic brain area. Single photon emission computed tomography reveals transient focal areas of decreased uptake consistent with the clinical symptoms. It is possible that pseudomigraine with temporary neurologic symptoms and lymphocytic pleocytosis could result from an activation of the immune system secondary to a recent viral infection, which would produce antibodies against neuronal or vascular antigens. This autoimmune attack may induce an aseptic leptomeningeal vasculitis, accounting for the headache and the transient symptoms likely through a spreading depression-like mechanism. PMID- 12720606 TI - The initial drug treatment of older patients with Parkinson's disease - consider an agonist, but don't demonise dopa. PMID- 12720607 TI - The new imperative of long-term care. PMID- 12720608 TI - Experience assessment and management of pain in people with dementia. AB - Pain is an inherently subjective experience that is difficult to prove. In a cognitively impaired older person whose verbal fluency is declining, both the experience and expression of pain are altered. Assessment poses many difficulties. Consequently the older person with dementia and pain may be under treated and poorly managed. This review addresses each of these issues and makes recommendations for more effective care in the future. The search strategy for this review was carried out using Medline (1990-2002), Embase (1989-2001) and ClinPSYCH (1990-2001) databases. References cited within these sources were also reviewed. Searches were limited to English language studies. The quality of relevant studies retrieved was assessed and information from relevant papers synthesised using narrative summary. PMID- 12720609 TI - A new classification of higher level gait disorders in patients with cerebral multi-infarct states. AB - BACKGROUND: cerebral multi-infarct states may lead to gait disorders in the absence of cognitive impairment. Where these gait disorders occur in the absence of neurological signs they have been termed gait apraxia or more recently higher level gait disorders. In this paper we hypothesise three main types based on presumptive sites of anatomical damage: (a) Ignition Apraxia, where damage is predominantly in the supplementary motor area and its connections, with good responses to external clues; (b) Equilibrium Apraxia, where damage is predominantly in the pre-motor area in its connections, with poor responses to external cues and (c) Mixed Gait Apraxia. SUBJECTS: the clinical features and measured gait parameters of 13 patients with cerebral multi-infarct states and higher-level gait disorder are described (7 with Ignition Apraxia and 6 with Equilibrium Apraxia) along with those of 6 healthy elderly control subjects. METHODS: baseline gait characteristics were assessed on a walkway, which measured the following: step lengths, width of base and velocity. RESULTS: measured baseline gait parameters support the above hypothesis. CONCLUSIONS: it is suggested, though not proven, that patients with Ignition Apraxia could have problems with internal cueing due to lesions in the supplementary motor area or its connections whereas those with Equilibrium Apraxia could have dysfunction predominantly in the pre-motor area and its connections. PMID- 12720610 TI - Medical conditions as risk factors for pressure ulcers in an outpatient setting. AB - OBJECTIVES: the purpose of this study was to evaluate the likelihood that the presence of certain medical conditions in older ambulatory patients are associated with the risk of developing a new pressure ulcer. DESIGN: a cohort study. SETTING AND SUBJECTS: a large outpatient record database from the United Kingdom called the General Practice Research Database. METHODS: the frequency of disease was reported as simple percentages and the associations between the medical conditions and the development of a pressure ulcer as instantaneous rate ratios. RESULTS: we studied 75,168 older individuals. Pressure ulcers occurred in 1,211 individuals. The medical conditions that were significantly associated with the development of a pressure ulcer after adjustment were: Alzheimer's disease, congestive heart failure, chronic obstructive pulmonary disease, cerebral vascular accident, diabetes mellitus, deep venous thrombosis, hip fracture, hip surgery, limb paralysis, lower limb oedema, malignancy, malnutrition, osteoporosis, Parkinson's disease, rheumatoid arthritis, and urinary tract infections. Angina, hypertension, and pneumonia were inversely associated with the development of a pressure ulcer. CONCLUSIONS: it is important that physicians recognise that patients with many medical conditions may be at higher risk for pressure ulcers so that even in the ambulatory care environment appropriate prevention and detection strategies can be directed towards the patients who are most likely to benefit. PMID- 12720612 TI - Dietary selenium intake affects cardiac susceptibility to ischaemia/reperfusion in male senescent rats. AB - BACKGROUND: cardiovascular ageing is associated with an increase in cardiac susceptibility to ischaemia and reperfusion. This has been suggested to be partly related to an increased sensitivity of the myocardium to the reactive oxygen species that are produced during post-ischaemic reperfusion. The aim of the present study was therefore to determine whether increasing cardiac glutathione peroxidase activity by a selenium-enriched diet could afford some protection against ischaemia and reperfusion to senescent rat hearts. METHODS: 22 months old male Wistar rats received either a high-selenium (1.5 mg Se/kg diet) or a low selenium (0.05 mg Se/kg diet) diet for 10 weeks. At the end of the diet, hearts were submitted to ischaemia and reperfusion ex vivo and either fixed for semi quantitative analysis of ultrastructural damage by electron microscopy or used for glutathione peroxidase activity assessment. RESULTS: high-selenium supply increased cardiac total, mitochondrial and cytosolic glutathione peroxidase activities. Moreover, this diet induced a significant improvement of cardiac post ischaemic functional recovery. Finally, this preservation of cardiac function was associated with a significant limitation of ultrastructural alterations of sarcomeres and mitochondria. CONCLUSION: our high-selenium diet considerably limits the sensitivity of senescent rat hearts to ischaemia and reperfusion. This finding suggests that peroxides might play a key role in the increase in cardiac sensitivity to ischaemia and reperfusion during ageing. Together with the observation that selenium status decreases with age in humans, our results indicate that reinforcing selenium supply could improve the prognosis of cardiovascular diseases in old patients. PMID- 12720611 TI - Long-standing and limiting long-standing illness in older people: associations with chronic diseases, psychosocial and environmental factors. AB - OBJECTIVE: to examine the associations between domains of chronic diseases, social, psychological and environmental factors and long-standing and limiting long-standing illness among older people. DESIGN: cross sectional survey. SETTING: national sample living in private households. SUBJECTS: 999 adults aged 65 years and over, mean age 73.2 years. MAIN OUTCOME MEASURES: self-reports of long-standing illness and limiting long-standing illness. RESULTS: the prevalence of long-standing illness was 61.8% (95% CI 58.8, 64.9) and that for limiting long standing illness was 40.0 (95% CI 38.0, 43.0). Strong associations between long standing illness and circulatory disease, odds ratio: 2.23 (95% CI 1.63, 3.05) and musculoskeletal disorders, odds ratio: 3.21 (95% CI 2.35, 4.39) were found. In addition associations with other domains were observed. For example, feelings of vulnerability, odds ratio: 1.79 (95% CI, 1.28, 2.51) from the psychological domain and, having close relatives living close by, odds ratio: 1.52 (95% CI 1.11, 2.09) from the social domain. CONCLUSION: the importance of considering a wide range of domains of human experience in the causation of limitations in society is emphasised. The currently dominant disease oriented view is insufficient to explain people's reported long-standing illness and limiting long standing illness. PMID- 12720614 TI - Developing the KAMA instrument (knowledge and management of abuse). AB - OBJECTIVE: to develop and validate an instrument which measures applied knowledge and practice regarding identification and management of potentially abusive situations in staff who work with older vulnerable inpatients. One of the anticipated uses will be to measure any change produced by attending an educational intervention designed to improve management of abuse. A vignette based instrument was chosen to measure applied knowledge and practice over other methods of assessments which were felt not to be practical or provide too much cueing for the volunteering staff. METHOD: a parallel form vignette-based instrument was developed so that not only baseline knowledge but also change in knowledge could be measured. Abusive scenarios were adapted from researchers' clinical practice and from the literature. Advice from senior nurses about the suitability of the vignettes for ward and community based staff was obtained and the instrument was piloted. Staff working with older people and employed by health or social services working in inner London completed the questionnaire. RESULTS: 79 (92%) of the eligible staff took part in this study. All completed the instrument. Version A of the Knowledge and Management instrument was answered by 39 staff, whilst 40 answered version B. The proportion of qualified and unqualified staff as well as the proportion employed by health or social services was similar for each parallel version of the instrument. The measure of internal consistency Cronbach's alpha was 0.89 for version A and 0.79 for version B. Parallel form reliability coefficient was calculated as 0.82, indicating that the two versions of the Knowledge and Management instrument perform similarly. Measures for concurrent validity according to years of experience and professional education also proved significant. Inter-rater reliability was calculated at 0.98 (P=0.01). Test-retest reliability at 0.69 (P=0.01) indicated that the instrument is stable yet sensitive to change. CONCLUSION: this instrument is a valid assessment tool that can be used to identify gaps in applied knowledge and management in potentially abusive situations in older people and allow knowledge in this field to be built on. PMID- 12720613 TI - Identification of registered nursing care of residents in English nursing homes using the Minimum Data Set Resident Assessment Instrument (MDS/RAI) and Resource Utilisation Groups version III (RUG-III). AB - AIM: to determine if a combination of Minimum Data Set/Resident Assessment Instrument (MDS/RAI) assessment variables and the Resource Utilisation Groups version III (RUG-III) case-mix system could be used as a method of identifying and reimbursing registered nursing care needs in long-term care. METHOD: the sample included 193 nursing home residents from four nursing homes from three different locations and care providers in England. The study included assessments of residents' care needs using either the MDS/RAI assessments or RUG stand-alone questionnaires and a time study that recorded the amount of nursing time received by residents over a 24-h period. Validity of RUG-III for explaining the distribution of care time between residents in different RUG-III groups was tested. The difference in direct and indirect care provided by registered general nurses (RGN) and care assistants (CA) to residents in RUG-III clinical groups was compared. RESULTS: the RUG-III system explained 56% of the variance in care time (Eta2, P=0.0001). Residents in RUG-III groups associated with particular medical and nursing needs (enhanced RGN care) received more than twice as much indirect RGN care time (t-test, P<0.001) and 1.4 times as much direct RGN and direct CA time (t-test, P<0.01) than residents with primarily cognitive impairment or physical problems only (standard RGN care). Residents with enhanced RGN care received an average of 48.1 min of RGN care in 24 h (95% CI 4.1-55.2) compared with an average of 31.1 min (95% CI 26.8-35.5) for residents in the standard RGN care group. A third low RGN care group was created following publication of the Department of Health guidance on NHS Funded Nursing Care. With three levels, the enhanced care group receives about 38% more than the standard group, and the low group receives about 50% of the standard group. CONCLUSIONS: the RUG-III system effectively differentiated between nursing home residents who are receiving 'low', 'standard' and 'enhanced' RGN care time. The findings could provide the basis of a reimbursement system for registered nursing time in long-term care facilities in the UK. PMID- 12720616 TI - Acquisition and short-term retention of inhaler techniques require intact executive function in elderly subjects. AB - BACKGROUND: patients with dementia are almost invariably unable to use any form of inhaler. Some elderly patients are unable to learn to use a metered dose inhaler or Turbohaler despite a normal abbreviated mental test score. Studies have shown that in many people this is due to unrecognised cognitive impairment and/or dyspraxia. The executive domains of cognition are particularly important in planning and sequencing; it might be expected therefore that disordered frontal (executive) function could be a predictor of poor inhaler technique in subjects with no overt features of dementia. OBJECTIVE: to explore the relationship between cognitive, and executive, function and the ability to acquire metered dose inhaler and Turbohaler technique in old age. DESIGN: a prospective randomised observational study with blinded evaluation. SUBJECTS: 30 inhaler-naive inpatients (21 female) with a mean age of 85 (range 75-94) and having a normal (8-10) abbreviated mental test score. METHODS: subjects received standardised metered dose inhaler and Turbohaler training and were scored on an analogue scale (for metered dose inhaler) or for competence (Turbohaler) the following day. The Mini-Mental State Examination and EXIT25 (for executive function) were performed by separate observers. RESULTS: significant correlation was found between the metered dose inhaler score and Mini-Mental State Examination (r 0.540, P<0.002) and EXIT25 (r -0.702, P<0.0001). Threshold effects emerged for the metered dose inhaler in that 18/19 with a competent score compared to 2/11 scored as incompetent had a Mini-Mental State Examination of >23 (P<0.01) and 19/19 compared to 0/11 had an EXIT25 of <15 (P<0.01). Similarly, for the Turbohaler 21/21 of the competent subjects had a Mini-Mental State Examination of >23 compared with 3/9 incompetent subjects (P<0.01), with 21/21 competent compared with 0/9 incompetent having an EXIT25 <15 (P<0.01). CONCLUSION: acquisition and short-term retention of metered dose inhaler and Turbohaler techniques is unlikely to be successful in frail elderly people who have an abnormal Mini-Mental State Examination and/or EXIT25 test. The latter test, when abnormal, is probably the superior predictor of inability to learn inhaler techniques. PMID- 12720615 TI - Using prescribing indicators to measure the quality of prescribing to elderly medical in-patients. AB - OBJECTIVES: to evaluate the performance of hospitals using eight indicators designed to assess prescribing practice in medical in-patients aged > or =65 years. DESIGN: local coalition teams were invited to collect cross-sectional prescribing and clinical data on 100 consecutive medical in-patients aged > or =65 years during a specific week in April 1999. SETTING: 102 hospitals across England. PARTICIPANTS: all NHS Trust hospitals in Wales and England were invited to participate in the study. MAIN OUTCOME MEASURES: the performance and inter hospital variation of hospitals in eight indicators of prescribing. Also, the age related appropriate use of anti-thrombotic stroke prophylaxis in atrial fibrillation, of aspirin in angina and of benzodiazepines. RESULTS: data were collected on 9,979 patients prescribed 70,458 medications. The number of hospitals achieving the prescribing goal for the indicators varied between 0 and 70. Frequency of administration instructions with 'as required' prescriptions were documented on 60% (10,403/17,258) of occasions. Generic (or acceptable proprietary) names were used for 84% (58,953/70,458) medications, 50% (4,870/9,778) of patients had documentation of allergy status on the drug chart and 23% (1,380/6,060) of patients had the potential risk of exceeding the maximum recommended dosage (4 g/24 h) of paracetamol. Long-acting hypoglycaemic drugs were prescribed to 50 patients. Anti-thrombotic stroke prophylaxis in atrial fibrillation were used appropriately for 53% (805/1,518) of patients, aspirin was used appropriately in angina for 90% (952/1,052) of patients and benzodiazepines were used appropriately for 49% (824/1,689) of patients. For the latter three indicators, the appropriate use of medications declined from 60% to 44%, 95% to 85% and 53% to 44% in patients aged >/=85 years compared with those aged 65-74 years. CONCLUSIONS: prescribing indicators were effective in evaluating the performance of 102 hospitals on prescribing practice to medical in-patients aged >/=65 years. Prescribing to elderly medical in-patients is sub-optimal but targets were achieved by some hospitals. This should inspire those hospitals not achieving high standards to improve their performance. The higher level of inappropriate prescribing with increasing age is unacceptable. PMID- 12720617 TI - Health status and assessed need for a cohort of older people admitted to nursing and residential homes. AB - OBJECTIVE: to investigate dependency and health status of a cohort of older people admitted for long term nursing or residential care and to compare these findings with assessments conducted by social services departments prior to placement. DESIGN: retrospective cohort study. SETTING: residential, nursing and dual registered homes within Nottingham Health Authority boundaries. SUBJECTS: 205 residents placed over 3 months. MAIN OUTCOME MEASURES: levels of disability, cognitive impairment and behavioural disturbance identified by assessment before and after admission. RESULTS: cognitive impairment and physical disability were significantly higher in nursing homes, although a third of residents in residential care had substantial physical disability. In nursing homes, a quarter of residents had low dependency needs but these had greater cognitive impairment than those in residential homes with the same level of dependency. Most residents had some degree of behavioural disturbance (particularly in nursing homes) and more severe disturbance was associated with greater cognitive impairment and more depressed mood, but not physical disability. A moderate level of agreement was found between preadmission and follow-up assessments of health status. CONCLUSIONS: a case-mix which includes higher dependency residents in residential homes and lower dependency residents in nursing homes is likely to reflect changes in the health status of residents following placement but also suggests that a range of placement criteria were used together, rather than individual indicators of need. Although pre-placement measures of disability and dependency were supported by follow-up assessments, it is essential that the needs of residents in long-term care are adequately monitored and managed, in particular those in residential care with higher dependency needs. PMID- 12720618 TI - An evaluation of footwear worn at the time of fall-related hip fracture. AB - BACKGROUND: a range of footwear features have been shown to influence balance in older people, however, little is known about the relationships between inappropriate footwear, falls and hip fracture. OBJECTIVES: to describe the characteristics of footwear worn at the time of fall-related hip fracture and establish whether the features of the shoe influenced the type of fall associated with the fracture. METHODS: 95 older people (average age 78.3 years, SD 7.9) who had suffered a fall-related hip fracture were asked to identify the footwear they were wearing when they fell. Footwear characteristics were then evaluated using a standardised assessment form. Information was also collected on the type and location of fall. RESULTS: the most common type of footwear worn at the time of the fall was slippers (22%), followed by walking shoes (17%) and sandals (8%). Few subjects were wearing high heels when they fell (2%). The majority of subjects (75%) wore shoes with at least one theoretically sub-optimal feature, such as absent fixation (63%), excessively flexible heel counters (43%) and excessively flexible soles (43%). Subjects who tripped were more likely to be wearing shoes with no fixation compared to those who reported other types of falls [chi(2)=4.21, df=1, P=0.033; OR=2.93 (95%CI 1.03-8.38)]. CONCLUSIONS: many older people who have had a fall-related hip fracture were wearing potentially hazardous footwear when they fell. The wearing of slippers or shoes without fixation may be associated with increased risk of tripping. Prospective studies into this proposed association appear warranted. PMID- 12720619 TI - Identifying a cut-off point for normal mobility: a comparison of the timed 'up and go' test in community-dwelling and institutionalised elderly women. AB - BACKGROUND: physical mobility testing is an essential component of the geriatric assessment. The timed up and go test measures basic mobility skills including a sequence of functional manoeuvres used in everyday life. OBJECTIVES: to create a practical cut-off value to indicate normal versus below normal timed up and go test performance by comparing test performance of community-dwelling and institutionalised elderly women. SETTING AND PARTICIPANTS: 413 community-dwelling and 78 institutionalised mobile elderly women (age range 65-85 years) were enrolled in a cross-sectional study. MEASUREMENTS: timed up and go test duration, residential and mobility status, age, height, weight and body mass index were documented. RESULTS: 92% of community-dwelling elderly women performed the timed up and go test in less than 12 seconds and all community-dwelling women had times below 20 seconds. In contrast only 9% of institutionalised elderly women performed the timed up and go test in less than 12 seconds, 42% were below 20 seconds, 32% had results between 20 and 30 seconds and 26% were above 30 seconds. The 10(th)-90(th) percentiles for timed up and go test performance were 6.0-11.2 seconds for community-dwelling and 12.7-50.1 seconds for institutionalised elderly women. When stratifying participants according to mobility status, the timed up and go test duration increased significantly with decreasing mobility (Kruskall-Wallis-test: p<0.0001). Linear regression modelling identified residential status (p<0.0001) and physical mobility status (p<0.0001) as significant predictors of timed up and go performance. This model predicted 54% of total variation of timed up and go test performance. CONCLUSION: residential and mobility status were identified as the strongest predictors of timed up and go test performance. We recommend the timed up and go test as a screening tool to determine whether an in-depth mobility assessment and early intervention, such as prescription of a walking aid, home visit or physiotherapy, is necessary. Community-dwelling elderly women between 65 and 85 years of age should be able to perform the timed up and go test in 12 seconds or less. PMID- 12720620 TI - Prevention of malnutrition in older people during and after hospitalisation: results from a randomised controlled clinical trial. AB - OBJECTIVE: to prevent the occurrence of weight loss during hospitalisation and following discharge by daily oral supplementation. DESIGN: in a prospective, randomised, controlled study of 80 patients aged 75 or more, and at risk of undernutrition based on their initial Mini Nutritional Assessment score, patients were randomised into a control group or one receiving oral supplementation. The intervention was a prescription of 200 ml sweet or salty sip feed twice daily (500 kcal, 21 g protein per day) throughout hospitalisation and convalescence. Nutritional status was assessed at baseline and after 2 months using Mini Nutritional Assessment and body weight record. RESULTS: compliance with oral supplementation was good and daily extra energy intake was 407+/-184 kcal. On day 60, significant weight loss from upon admission was observed in the control group (-1.23+/-2.5 kg; P=0.01), but not in the supplemented group (0.28+/-3.8 kg; NS). At the end of the study, Mini Nutritional Assessment scores were higher in the supplemented group than in the control group (23.5+/-3.9 versus 20.8+/-3.6; P<0.01). CONCLUSION: use of daily oral supplementation during and after hospitalisation maintains body weight and increases Mini Nutritional Assessment score in patients at risk of undernutrition. PMID- 12720621 TI - Impact of age on QT interval and QT dispersion in healthy subjects: a regression analysis. AB - OBJECTIVE: although the isolated effects of age on QT interval and QT dispersion (QTd) have been previously investigated, no data are available on the simultaneous effects of age and other physiological or lifestyle factors on QT interval and QTd in healthy subjects. We studied the effects of age, gender, body mass index, smoking status, and blood pressure on these electrocardiographic parameters. DESIGN: observational study. SETTING: academic medical centre. PARTICIPANTS AND MEASUREMENTS: age, gender, body mass index, smoking status, and blood pressure were obtained from 191 consecutive healthy subjects (101 males and 90 females, age range 19-89 years). The subjects were divided into three groups according to their age: <30 (n=56), 30-65 (n=49), and >65 years (n=86). RESULTS: heart-rate corrected QT interval (QTc, Bazett's formula) progressively increased with advancing age (389+/-3 vs. 411+/-4 vs. 418+/-3 ms, means+/-SEM; P<0.01). By contrast, no differences in QTd were observed across the three groups (36+/-2 vs. 35+/-3 vs. 40+/-2 ms, P=NS). A multivariate regression analysis showed that age (P<0.01) and body mass index (P=0.04) independently predicted QT interval while gender was a weak (P=0.09) predictor of QTd. CONCLUSIONS: after adjusting for gender, smoking status, and blood pressure, age and body mass index independently predicted QT interval in healthy subjects. By contrast, age is not a predictor of QTd. The increase of QT interval associated with ageing and body mass index might be secondary to cardiac hypertrophy and myocardial action potential prolongation. PMID- 12720622 TI - Nursing home closures: effects on capacity and reasons for closure. AB - OBJECTIVES: to identify the rate of closure of nursing homes for older people, the types of homes closing and the reasons for closure. DESIGN: mixed method study including a census and telephone survey of registration and inspection units and interviews with independent providers. PARTICIPANTS: 81 of 96 health authority and joint registration and inspection unit managers in England completed the census and 39 managers participated in a further telephone survey. Twenty-five independent providers were interviewed. RESULTS: closures resulted in a net loss of 6% of nursing homes and 4.9% of nursing places during 2000-2001. Smaller homes were more likely to close and were increasingly seen as unviable. The majority of closed homes were reported to have provided good quality care. Shortages of nursing staff were of widespread concern. The dominant combination of factors identified by providers was low fees and concerns about the cost implications of the new care standards. Changes in demand were reflected in the placement of high dependency residents in residential rather than nursing homes. CONCLUSIONS: in the absence of policy interventions capacity will continue to reduce, with smaller homes most likely to disappear. There is an urgent need to address the supply and efficient use of nursing staff skills in care homes. While fee levels are the primary concern the effect of the proposed care standards was clearly having an effect. Even with subsequent amendment to these standards, unless authorities use capacity funding to raise fees and improve expectations, providers are likely to continue to exit the market. PMID- 12720624 TI - Performance indicators for hospital services for older people. AB - BACKGROUND: standardised assessment is recommended in geriatric practice to improve patient care and generate quality data for audit and research. High level indicators used to measure performance of rehabilitation units suggested that more patients were discharged to long-term residential care from one of two hospitals in a Health Authority in South East England. OBJECTIVES: to test whether the information provided by standardised assessment could inform performance indicators used to compare outcomes between hospitals. DESIGN: prospective observational study. SUBJECTS: consecutive patients admitted for rehabilitation to two general geriatric rehabilitation wards in each of two nearby district general hospitals. METHODS: patients were assessed using standard tools (Barthel and Abbreviated Mental Test score) and various scales of the interRAI MDS assessment system (mental and physical functioning, pressure ulcers, continence, falls, mood), within a week of admission and up to one week before discharge. Place of residence prior to admission and discharge destination were determined. RESULTS: on bivariate analysis there was a significant difference in discharge to residential and nursing homes between hospitals. Results from multivariate logistic regression analysis showed an increased risk for institutionalisation at discharge for women [odds ratio 2.42 (95% CI 1.41-4.14)] and patients with impaired cognitive function [odds ratio 1.53 (95% CI 1.28-1.82) for each point increase in MDS cognitive performance scale] and physical function [odds ratio 1.15 (95% CI 1.08-1.22) for each point increase in MDS short ADL scale]. Barthel and Abbreviated Mental Test showed similar odds ratios. Hospital did not remain a significant predictor of discharge destination following adjustment for patients' physical and cognitive function. CONCLUSIONS: comparisons of outcomes between hospitals could be misleading unless informed by standardised data on physical and mental functioning of rehabilitation patients. PMID- 12720623 TI - Characteristics of early fallers on elderly patient rehabilitation wards. AB - BACKGROUND: falls are one of the most frequent complications on rehabilitation wards for elderly patients. OBJECTIVE: to study the characteristics of early and late fallers. DESIGN: prospective observational study with blinded end-point evaluation. SETTING: a non-acute geriatric hospital. SUBJECTS: 1025 consecutive patients admitted for rehabilitation following treatment for an acute illness. MAIN OUTCOME MEASURES: early/late fallers and time to first fall from admission. RESULTS: we identified 824 non-fallers and 201 fallers. Seventy seven (38.3%) fell during the first week. The incidence decreased progressively in subsequent weeks. Early fallers were more likely to have a past history of falls (P=0.0009), an unsafe gait (P=0.001), confusion (P<0.0001) and be admitted from medical wards (P=0.03). Patients admitted from orthopaedic wards having sustained a lower limb fracture were significantly less likely to have an early fall compared to all other patients (P=0.027). When compared to later fallers, early fallers were more likely to have a past history of falls (P=0.045). They were less likely to be admitted from an orthopaedic ward (P=0.01) or to have sustained a fracture of the lower limbs (P=0.002). Logistic regression analysis showed that a past history of falls, confusion and an unsafe gait were independent risk factors predisposing to early falls. The Kaplan-Meier survival analysis showed a significantly higher cumulative risk of falling associated with these characteristics. CONCLUSION: the initial week of patients' rehabilitation is associated with the greatest risk of falling. Early fallers can be predicted by easily identifiable characteristics. This highlights the need for early fall risk assessment. PMID- 12720625 TI - Defecography in symptomatic older women living at home. AB - BACKGROUND: complaints of defecation disorders in older patients living at home is an emerging problem. Little is known about radiological examination of this population. OBJECTIVE: this study aimed to analyse the yield of defecography in women older than 75 years, living at home and complaining of defecation disorders. DESIGN AND SETTINGS: prospective study of patients referred to a radiology department in a tertiary-care medical centre in Rouen, France. SUBJECTS AND METHODS: 52 women (mean age: 78, range: 75-93) complaining of constipation, faecal incontinence or pelvic pain underwent defecography. Defecography was performed after intake of a barium meal and vaginal opacification. Radiographs were analysed accordingly with the established criteria. RESULTS: defecography showed perineal descent in 27 patients, rectocele in 29, intussusception in 33 and enterocele in 14. A combination of abnormalities was found in 40 women. Only 3 studies were normal. There was no significant association between symptoms and pelvic disorders revealed by defecography. CONCLUSIONS: defecography in symptomatic women aged 75 years and over did not raise any technical difficulty. It revealed a 77% rate of abnormalities, but there was no relationship between the symptoms and the detected abnormalities. PMID- 12720626 TI - A difficult case of inflammatory myositis. AB - A 79-year-old lady presented with subclinical autoimmune hypothyroidism and proximal muscle weakness, which failed to respond to thyroxine. Subsequent electromyography and muscle biopsy confirmed polymyositis, which responded well to immunosuppressive therapy. She presented 4 months later with a thyroid carcinoma, most likely anaplastic, and died shortly afterwards. Although the association between malignancy and polymyositis remains controversial, to our knowledge this is the first report of thyroid carcinoma occuring in association with polymyositis and autoimmune hypothyroidism. PMID- 12720627 TI - Newly diagnosed HIV infection in an octogenarian: the elderly are not 'immune'. AB - CASE REPORT: an 83-year-old heterosexual man with malaise tested positive for HIV infection antibodies and has responded well to triple antiretroviral therapy. DISCUSSION: HIV should not be discounted on the basis of age when an elderly patient presents for the investigation of illness. PMID- 12720629 TI - False positive findings in the ultrasound assessment of postvoid residual urine volume. PMID- 12720630 TI - Hospital readmission for elderly patients: subjective versus objective considerations. PMID- 12720631 TI - Photodynamic therapy in age-related macular degeneration: missing information. PMID- 12720632 TI - Reliability of blood pressure measurement for orthostatic hypotension: a hospital based questionnaire surve. PMID- 12720633 TI - Clinical assessment of symptomatic osteoarthritis in older people. PMID- 12720634 TI - An audit of outcomes in day hospital-based crisis intervention. PMID- 12720635 TI - Long waiting list for by-pass surgery. PMID- 12720636 TI - Randomised controlled trial of a nurse-led falls prevention programme. PMID- 12720640 TI - Should this study change my practice? PMID- 12721092 TI - Synergies at the synapse. PMID- 12721093 TI - Crosstalk between presynaptic angiotensin receptors, bradykinin receptors and alpha 2-autoreceptors in sympathetic neurons: a study in alpha 2-adrenoceptor deficient mice. AB - 1. In mouse atria, angiotensin II and bradykinin lose much or all of their noradrenaline release-enhancing effect when presynaptic alpha(2)-autoinhibition does not operate either because of stimulation with very brief pulse trains or because of treatment with alpha(2) antagonists. We now studied this operational condition in alpha(2)-adrenoceptor-deficient mice. Release of (3)H-noradrenaline was elicited by electrical stimulation. 2. In tissues from wild-type (WT) mice, angiotensin II and bradykinin increased the overflow of tritium evoked by 120 pulses at 3 Hz. This enhancement did not occur or was much reduced when tissues were stimulated by 120 pulses at 3 Hz in the presence of rauwolscine and phentolamine, or when they were stimulated by 20 pulses at 50 Hz. 3. In tissues from mice lacking the alpha(2A)-adrenoceptor (alpha(2A)KO) or the alpha(2B) adrenoceptor (alpha(2B)KO), the concentration-response curves of angiotensin II and bradykinin (120 pulses at 3 Hz) were unchanged. In tissues from mice lacking the alpha(2C)-adrenoceptor (alpha(2C)KO) or both the alpha(2A)- and the alpha(2C) adrenoceptor (alpha(2AC)KO), the concentration-response curves were shifted to the same extent downwards. 4. As in WT tissues, angiotensin II and bradykinin lost most or all of their effect in alpha(2A)KO and alpha(2AC)KO tissues when rauwolscine and phentolamine were present or trains consisted of 20 pulses at 50 Hz. 5. Rauwolscine and phentolamine increased tritium overflow evoked by 120 pulses at 3 Hz up to seven-fold in WT and alpha(2B)KO tissues, three-fold in alpha(2A)KO and alpha(2C)KO tissues, and two-fold in alpha(2AC)KO tissues. 6. Results confirm that angiotensin II and bradykinin require ongoing alpha(2) autoinhibition for the full extent of their release-enhancing effect. Specifically, they require ongoing alpha(2C)-autoinhibition. The peptide effects that remain in alpha(2C)-autoreceptor-deficient mice seem to be because of alpha(2B)-autoinhibition. The results hence also suggest that in addition to alpha(2A)- and alpha(2C)- mouse postganglionic sympathetic neurons possess alpha(2B)-autoreceptors. PMID- 12721094 TI - Endothelin as a causative factor of blunted volume reflex in diabetic rats. AB - 1. The study investigated whether endothelin (ET) contributed to the diabetes associated alterations in volume reflex and characterised the receptor subtype that might be involved. The influence of renal sympathetic nerves on these aspects of ET was also examined. 2. Groups of nondiabetic and streptozotocin induced diabetic rats were subjected to an acute isotonic saline volume expansion (VE), 10% body wt in the presence and absence of ET antagonists. 3. Cumulative urine sodium excretion (CuU(Na)V) after VE in diabetic rats reached values of 116+/-10 in the denervated and 74+/-6 micromol min(-1) g kidney wt(-1) in the innervated kidneys, which were both less (both P<0.001) than those achieved in the nondiabetic rats, at 267+/-9 in the denervated and 183+/-10 micromol min(-1) g kidney wt(-1) in the innervated kidney, respectively. 4. Diabetic rats pretreated with a nonselective ET(A)/ET(B) antagonist had an enhanced CuU(Na)V in the denervated kidneys by 37% (P<0.01) compared to that of untreated diabetic rats. At both doses of SB209670 these increments were less than the values obtained previously in nondiabetic rats (both P<0.01). The ET(A)/ET(B) antagonist had no meaningful effect on CuU(Na)V in the innervated kidneys of the diabetic rats, whereas previous studies in nondiabetic rats showed the response to be depressed. The CuU(Na)V responses to VE in diabetic rats given the selective ET(A) antagonist were not different from those observed in untreated diabetic rats, irrespective of whether or not the renal nerves were present. In nondiabetic rats, the ET(A) antagonist had an action similar to the mixed antagonist. 5. These findings demonstrate that activation of ET(B) receptors contributes to the depressed ability to excrete a saline load in diabetes mellitus, but its impact is obscured by the influence of the renal nerves. PMID- 12721096 TI - Ethanol tachyphylaxis in spinal cord motorneurons: role of metabotropic glutamate receptors. AB - 1. Ethanol (EtOH) tachyphylaxis (acute tolerance), a time-dependent decrease in apparent potency, is known in vivo and in some neuronal preparations. The present studies characterize EtOH tachyphylaxis in spinal motorneurons and test the hypothesis that metabotropic glutamate receptors (mGluRs) play a role. 2. Patch clamp studies were carried out in motorneurons in rat spinal cord slices. Currents were evoked by pulses of glutamate, alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic acid (AMPA) or N-methyl-D-aspartic acid (NMDA). 3. In nine of 15 cells, ethanol depression of glutamate-evoked currents was time dependent. EtOH depressed current area 36.9+/-3% at 8-10 min, but only 16.8+/-3% at 20 min. Mean reduction in depression was 20.1+/-1%, N=9. Tachyphylaxis was less prominent in currents evoked by AMPA or NMDA, appearing in two of 10 AMPA and three of 11 NMDA currents. 4. The mGluR agonist trans-(1S,3R)-1-amino-1,3 cyclopentanedicarboxylic acid (ACPD) increased, the antagonist (+/-)-alpha-methyl 4-carboxyphenylglycine (MCPG) decreased the area of glutamate-evoked currents. ACPD also increased the area of NMDA- and AMPA-evoked currents. 5. ACPD increased the incidence of tachyphylaxis in glutamate-evoked currents to 100% (N=9); MCPG markedly reduced tachyphylaxis. ACPD also increased the incidence of tachyphylaxis in currents evoked by NMDA and AMPA to five of eight and four of seven neurons, respectively. 6. Block of G-protein pathways by intracellular GDP beta-s abolished tachyphylaxis in glutamate-evoked currents (N=8); however, currents recovered only partially following EtOH washout. 7. Activation of mGluRs contributes to neuronal tachyphylaxis to EtOH in spinal cord motorneurons, probably via G-protein pathways. PMID- 12721095 TI - Growth hormone protects human lymphocytes from irradiation-induced cell death. AB - 1. Undesired effects of cancer radiotherapy mainly affect the hematopoietic system. Growth hormone (GH) participates in both hematopoiesis and modulation of the immune response. We report both r-hGH cell death prevention and restoration of secretory capacities of irradiated human peripheral blood lymphocytes (PBL) in vitro. 2. r-hGH induced cell survival and increased proliferation of irradiated cells. Western blot analysis indicated that these effects of GH were paralleled by increased expression of the antiapoptotic protein Bcl-2. 3. r-hGH restored mitogen-stimulated release of IL-2 by PBL. Preincubation of irradiated lymphocytes with the growth hormone receptor (GHR) antagonists B2036 and G120 K abrogated r-hGH-dependent IL-2 release. 4. These results demonstrate that r-hGH protects irradiated PBL from death in a specific, receptor-mediated manner. Such effect of r-hGH on PBL involves activation of the antiapoptotic gene bcl-2 and prevention of cell death, associated with preserved functional cell capacity. Finally, potential use of GH as an immunopotentiating agent could be envisioned during radiation therapy of cancer. PMID- 12721097 TI - Cannabinoid CB(1) receptor-mediated inhibition of hippocampal acetylcholine release is preserved in aged mice. AB - 1. The cannabinoid CB(1) receptor inverse agonist/antagonist SR 141716 increases acetylcholine release in rodent hippocampus and improves memory in some experimental paradigms. Since drugs like SR 141716 may represent a novel class of cognition-enhancing drugs, we wanted to check whether the function of the CB(1) receptor is preserved during ageing. 2. Hippocampal and striatal slices from 2- to 3- and 24- to 28-month-old C57BL/6J mice were preincubated with [(3)H]-choline or [(3)H]-noradrenaline ([(3)H]-NA) and superfused. 3. The cannabinoid receptor agonist WIN 55212-2 inhibited, and SR 141716 facilitated, the electrically (3 Hz) evoked tritium overflow in hippocampal slices (preincubated with [(3)H]-choline) from young and aged mice to the same extent. The evoked overflow per se was less by 33% in slices from aged animals. 4. WIN 55212-2 and SR 141716 did not affect, but the muscarinic receptor agonist oxotremorine inhibited, the evoked (3 Hz) overflow in striatal slices (preincubated with [(3)H]-choline) from young and aged mice to the same extent. The evoked overflow per se tended to be less in slices from aged animals. 5. The evoked (0.3 Hz) overflow in hippocampal slices (preincubated with [(3)H]-NA) was not affected by WIN 55212-2 and SR 141716, but was inhibited by histamine (via H(3) receptors) in slices from young mice and, to a somewhat less extent, in slices from aged mice. The evoked overflow per se did not differ between age groups. 6. In conclusion, the function of the CB(1) receptor involved in the tonic inhibition of hippocampal acetylcholine release is preserved in aged mice. PMID- 12721098 TI - Identification and functional characterization of hemorphins VV-H-7 and LVV-H-7 as low-affinity agonists for the orphan bombesin receptor subtype 3. AB - 1. The human orphan G-protein coupled receptor bombesin receptor subtype 3 (hBRS 3) was screened for peptide ligands by a Ca(2+)mobilization assay resulting in the purification and identification of two specific ligands, the naturally occurring VV-hemorphin-7 (VV-H-7) and LVV-hemorphin-7 (LVV-H-7), from human placental tissue. These peptides were functionally characterized as full agonists with unique specificity albeit low affinity for hBRS-3 compared to other bombesin receptors. 2. VV-H-7 and LVV-H-7 induced a dose-dependent response in hBRS-3 overexpressing CHO cells, as well as in NCI-N417 cells expressing the hBRS-3 endogenously. The affinity of VV-H-7 was higher in NCI-N417 cells compared to overexpressing CHO cells. In detail, the EC(50) values were 45+/-15 microM for VV H-7 and 183+/-60 microM for LVV-H-7 in CHO cells, and 19+/-6 microM for VV-H-7 and 38+/-18 microM for LVV-H-7 in NCI-N417 cells. Other hemorphins had no effect. Gastrin-releasing peptide (GRP) and neuromedin B (NMB) showed similar EC(50) values of 13-20 microM (GRP) and of 1-2 microM (NMB) on both cell lines. 3. Structure-function analysis revealed that both the N-terminal valine and the C terminal phenylalanine residues of VV-H-7 are critical for the ligand-receptor interaction. 4. Endogenous hBRS-3 in NCI-N417 activated by VV-H-7 couples to phospholipase C resulting in changes of intracellular calcium, which is initially released from an inositol trisphosphate (IP(3))-sensitive store followed by a capacitive calcium entry from extracellular space. 5. VV-H-7-induced hBRS-3 activation led to phosphorylation of p42/p44-MAP kinase in NCI-N417 cells, but did not stimulate cell proliferation. In contrast, phosphorylation of focal adhesion kinase (p125(FAK)) was not observed. PMID- 12721099 TI - Polarized Th1 and Th2 cells are less responsive to negative feedback by receptors coupled to the AC/cAMP system compared to freshly isolated T cells. AB - 1. The adenylyl cyclase (AC)/cyclic adenosine monophosphate (cAMP) system is known to negatively regulate transcriptional activity of T cells, thereby possibly modulating T-cell-mediated responses at the sites of inflammation. Effects of cAMP have been widely studied in freshly isolated T cells and T-cell clones; yet, effects in differentiated Th1 and Th2 cells are largely unknown. 2. To obtain differentiated T helper cells, we activated naive T cells for 1 week in the presence of IL-12 plus alpha-IL-4 to generate Th1-type cells and in the presence of IL-4 plus alpha-IL-12 to generate Th2-type cells. 3. We demonstrate that, in contrast to freshly isolated T cells, the production of Th1 (IFN-gamma) and Th2 (IL-4, IL-5) cytokines in polarized T helper cells is not strictly controlled by the activation of AC/cAMP-linked beta(2)-adrenergic and prostaglandin (PG)E(2) receptors. 4. In Th2 cells, PGE(2) could still activate the G(s) protein-coupled AC/cAMP system and subsequently induce CREB phosphorylation, whereas PGE(2) was unable to activate the cAMP-dependent pathway in Th1 cells. In both Th1 and Th2 cells, the induction of CREB phosphorylation by beta(2)-agonist fenoterol was impaired. 5. The loss of control over cytokine production by cAMP elevating agents in differentiated Th1 and Th2 subsets may have important implications for the regulation of Th1- and Th2-mediated diseases, in particular those associated with the ongoing immune responses. PMID- 12721101 TI - Ezetimibe potently inhibits cholesterol absorption but does not affect acute hepatic or intestinal cholesterol synthesis in rats. AB - 1. Ezetimibe (1-(4-fluorophenyl)-(3R)-[3-(4-fluorophenyl)-(3S)-hydroxypropyl] (4S)-(4-hydroxyphenyl)-2-azetidinone) and its analog SCH48461 are potent and selective cholesterol absorption inhibitors that inhibit the transport of cholesterol across the intestinal wall, thereby lowering plasma cholesterol. 2. After a dose response for ezetimibe in rats was established, experiments were conducted to determine whether acute administration could alter hepatic or intestinal cholesterol synthesis. To determine whether this class of intestinal cholesterol absorption inhibitors could discriminate between newly synthesized cholesterol in the intestine versus exogenously administered cholesterol, rats were intraduodenally dosed with (14)C-cholesterol and (3)H-mevalonate, and mesenteric lymph was analyzed for radiolabeled cholesterol and cholesteryl ester content. 3. Ezetimibe attenuated diet-induced hypercholesterolemia 60-94% at doses of 0.1-3 mg x kg(-1) in rats. A single administration of ezetimibe did not have a direct effect on intestinal or hepatic cholesterol synthesis, while ketoconazole significantly inhibited cholesterol synthesis after a single dose. The ezetimibe analog, SCH48461, inhibited the movement of exogenously administered cholesterol into lymph, but did not affect the appearance of newly synthesized cholesterol into lymph. 4. These data suggest that this class of cholesterol absorption inhibitors does discriminate by blocking the movement of exogenous cholesterol in the enterocyte before it reaches the intracellular cholesterol pool to be incorporated into intestinal lipoproteins, without affecting the incorporation of newly synthesized cholesterol into intestinal lipoproteins. PMID- 12721100 TI - Extracellular nucleotides induce vasodilatation in human arteries via prostaglandins, nitric oxide and endothelium-derived hyperpolarising factor. AB - 1. The present study was aimed at examining P2 receptor-mediated vasodilatation in human vessels. The isometric tension was recorded in isolated segments of the human left internal mammary artery branches precontracted with 1 microM noradrenaline. 2. Endothelial denudation abolished the dilator responses. 3. The selective P2Y(1) agonist, 2-MeSADP, induced a potent vasodilatation (pEC(50)=6.9+/-0.1). The P2Y(1) antagonist of 10 microM, MRS 2216, shifted the 2 MeSADP concentration-response curve 1.1 log units to the right. The combined P2Y(1) and P2X agonist, 2-MeSATP, stimulated a dilatation with a potency similar to that of 2-MeSADP. Furthermore, MRS 2216 had a similar antagonistic effect on both 2-MeSATP and 2-MeSADP indicating that P2X receptors do not mediate vasodilatation. 4. Both the P2Y(2/4) agonist, UTPgammaS and the P2Y(6) agonist, UDPbetaS, stimulated potent dilatations (pEC(50)=7.8+/-0.4 for UTPgammaS and 8.4+/-0.2 for UDPbetaS). 5. The 2-MeSADP-induced nitric oxide (NO)-mediated dilatation was studied in the presence of 10 micro M indomethacin, 50 nM charybdotoxin and 1 microM apamin. The involvement of the endothelium-derived hyperpolarising factor (EDHF) was investigated in the presence of 0.1 mM L-NOARG and indomethacin. The involvement of prostaglandins was investigated in the presence of L-NOARG, charybdotoxin and apamin. Both NO, EDHF and prostaglandins mediated 2-MeSADP dilatation with similar efficacy (E(max)=25+/-5% for NO, 25+/ 6% for EDHF and 27+/-5% for prostaglandins). 6. In conclusion, extracellular nucleotides induce endothelium-derived vasodilatation in human vessels by stimulating P2Y(1), P2Y(2/4) and P2Y(6) receptors, while P2X receptors are not involved. Endothelial P2Y receptors mediate dilatation by release of EDHF, NO and prostaglandins. PMID- 12721102 TI - Contribution of human cytochrome p-450 isoforms to the metabolism of the simplest phenothiazine neuroleptic promazine. AB - 1. The aim of the present study was to identify human cytochrome p-450 isoforms (CYPs) involved in 5-sulphoxidation and N-demethylation of the simplest phenothiazine neuroleptic promazine in human liver. 2. The experiments were performed in the following in vitro models: (A). a study of promazine metabolism in liver microsomes-(a). correlations between the rate of promazine metabolism and the level and activity of CYPs; (b). the effect of specific inhibitors on the rate of promazine metabolism (inhibitors: CYP1A2-furafylline, CYP2D6-quinidine, CYP2A6+CYP2E1-diethyldithiocarbamic acid, CYP2C9-sulfaphenazole, CYP2C19 ticlopidine, CYP3A4-ketoconazole); (B). promazine biotransformation by cDNA expressed human CYPs (Supersomes 1A1, 1A2, 2A6, 2B6, 2C9, 2C19, 2E1, 3A4); (C). promazine metabolism in a primary culture of human hepatocytes treated with specific inducers (rifampicin-CYP3A4, CYP2B6 and CYP2C inducer, 2,3,7,8 tetrachlordibenzeno-p-dioxin (TCDD)-CYP1A1/1A2 inducer). 3. In human liver microsomes, the formation of promazine 5-sulphoxide and N-desmethylpromazine was significantly correlated with the level of CYP1A2 and ethoxyresorufin O deethylase and acetanilide 4-hydroxylase activities, as well as with the level of CYP3A4 and cyclosporin A oxidase activity. Moreover, the formation of N desmethylpromazine was correlated well with S-mephenytoin 4'-hydroxylation. 4. Furafylline (a CYP1A2 inhibitor) and ketoconazole (a CYP3A4 inhibitor) significantly decreased the rate of promazine 5-sulphoxidation, while furafylline and ticlopidine (a CYP2C19 inhibitor) significantly decreased the rate of promazine N-demethylation in human liver microsomes. 5. The cDNA-expressed human CYPs generated different amounts of promazine metabolites, but the rates of CYP isoforms to catalyse promazine metabolism at therapeutic concentration (10 microM) was as follows: 1A1>2B6>1A2>2C9>3A4>2E1>2A6>2D6>2C19 for 5-sulphoxidation and 2C19>2B6>1A1>1A2>2D6>3A4>2C9>2E1>2A6 for N-demethylation. The highest intrinsic clearance (V(max)/K(m)) was found for CYP1A subfamily, CYP3A4 and CYP2B6 in the case of 5- sulphoxidation, and for CYP2C19, CYP1A subfamily and CYP2B6 in the case of N-demethylation. 6. In a primary culture of human hepatocytes, TCDD (a CYP1A subfamily inducer), as well as rifampicin (mainly a CYP3A4 inducer) induced the formation of promazine 5-sulphoxide and N desmethylpromazine. 7. Regarding the relative expression of various CYPs in human liver, the obtained results indicate that CYP1A2 and CYP3A4 are the main isoforms responsible for 5-sulphoxidation, while CYP1A2 and CYP2C19 are the basic isoforms that catalyse N-demethylation of promazine in human liver. Of the other isoforms studied, CYP2C9 and CYP3A4 contribute to a lesser degree to promazine 5 sulphoxidation and N-demethylation, respectively. The role of CYP2A6, CYP2B6, CYP2D6 and CYP2E1 in the investigated metabolic pathways of promazine seems negligible. PMID- 12721103 TI - Effects of flecainide and quinidine on Kv4.2 currents: voltage dependence and role of S6 valines. AB - 1. The effects of flecainide and quinidine were studied on wild-type Kv4.2 channels (Kv4.2WT), channels with deletion of the N-terminal domain (N-del) and channels with mutations in the valine residues located at positions 402 and 404 in the presence (V[402,404]I) or in the absence (N-del/V[402,404]I) of the N terminus. 2. The experiments were performed at 37 degrees C on COS7 cells using the whole-cell configuration of the patch-clamp technique. 3. Flecainide and quinidine inhibited Kv4.2WT currents in a concentration-dependent manner (IC(50)=23.6+/-1.1 and 12.0+/-1.4 microMat +50 mV, respectively), similar to their potency for the rest of the constructs at the same voltage. In Kv4.2WT channels, flecainide- and quinidine-induced block increased as channel inactivation increased. In addition, the inhibition produced by quinidine, but not by flecainide, increased significantly at positive test potentials. Similar effects were observed in N-del channels. However, in V[402,404]I and N del/V[402,404]I channels, the voltage dependence of block by both quinidine and flecainide was lost, without significant modifications in potency at +50 mV. 4. These results point to an important role for S6 valines at positions 402 and 404 in mediating voltage-dependent block by quinidine and flecainide. PMID- 12721104 TI - AT1 receptor antagonist therapy preferentially ameliorated right ventricular function and phenotype during the early phase of remodeling post-MI. AB - 1. The influence of AII on contractile dysfunction, regulation of the tyrosine kinase-dependent signaling molecule extracellular signal-regulated kinase (ERK), and natriuretic peptide gene expression were examined in the noninfarcted left ventricle (NILV) and right ventricle (RV) during the early phase of remodeling post-myocardial infarct (MI) in the rat. The selective AT(1) receptor antagonist irbesartan was administered <10 h following coronary artery ligation, and rats were killed either at 4-day or 2-week post-MI. 2. At 4 days post-MI, left ventricular systolic pressure (LVSP: sham=125+/-12, MI=91+/-4 mmHg) was decreased, whereas left ventricular end-diastolic pressure (LVEDP: sham=9+/-2, MI=17+/-2 mm Hg), right ventricular systolic (RVSP: sham=26+/-1, MI=34+/-2 mm Hg), and end-diastolic pressures (RVEDP: sham=3+/-0.5, MI=7+/-1 mm Hg) were increased. ERK phosphorylation was significantly elevated in the NILV and RV. 3. Irbesartan (40 mg x kg(-1)/day(-1)) administration did not improve left ventricular function, or suppress increased ERK phosphorylation in the 4-day post MI rat. By contrast, irbesartan therapy normalized RVSP (MI+irbesartan=25+/-1 mm Hg), RVEDP (MI+irbesartan=3+/-0.3 mm Hg), and reduced ERK1 (MI=3.0+/-0.6, MI+irbesartan=2.0+/-0.3-fold increase), and ERK2 (MI=3.8+/-0.8, MI+irbesartan=2.2+/-0.5-fold increase) phosphorylation. 4. In 2-week post-MI rats, biventricular dysfunction was associated with increased prepro-ANP, and prepro-BNP mRNA expression. Irbesartan therapy normalized RVSP, attenuated RVEDP, and abrogated natriuretic peptide mRNA expression (prepro-ANP; MI=9+/-2, MI+irbesartan=2+/-1-fold increase, prepro-BNP; MI=6+/-2, MI+irbesartan=1+/-1-fold increase), whereas both transcripts remained elevated in the NILV despite the partial attenuation of LVEDP. 5. These data suggest that the therapeutic benefit of irbesartan treatment during the early phase of remodeling post-MI was associated with the preferential amelioration of RV contractile function and phenotype. PMID- 12721105 TI - Characterization of a fluorescent conjugate of the rabbit angiotensin AT(1) receptor. AB - 1. The rabbit AT(1) receptor (AT(1)R) for angiotensin II (A(II)) has been conjugated to the yellow fluorescent protein (YFP) in order to establish the pharmacological profile of such a fusion protein and to facilitate the study of ligand-induced regulation. 2. A(II) bound AT(1)R-YFP (K(D) 8.1 nM in transiently transfected cells) and stimulated HEK 293 cells expressing the fusion protein at concentration ranges similar to the ones that stimulate the contraction of the isolated rabbit aorta. Antagonists found to be insurmountable in the latter assay (candesartan and EXP-3174 being the most extreme cases) were also insurmountable in the phospholipase A(2) assay applied to cells expressing AT(1)R-YFP, whereas losartan appeared to be surmountable in both assays. 3. Cells expressing AT(1)R YFP exhibited a membrane-associated fluorescence that was partly and reversibly translocated into intracellular structures upon A(II) stimulation (confocal microscopy); the nonpeptide antagonists were not active in this respect, but prevented the effect of the agonist. 4. A(II) treatment increased the quantity of the fusion protein in cells, and phorbol 12-myristate 13-acetate (PMA) treatment even more so (immunoblot, confocal microscopy) but, unlike the agonist, the latter drug did not induce receptor endocytosis. A protein kinase C (PKC) inhibitor prevented the effect of either A(II) or PMA on AT(1)R-YFP abundance. 5. The conjugate AT(1)R-YFP retains the pharmacological properties of the parent rabbit AT(1)R. Agonist-induced downregulation was not documented using this system; to the contrary, we have observed a PKC-mediated increased expression AT(1)R-YFP likely to be the result of a decreased breakdown rate of the fusion protein. PMID- 12721106 TI - Effects of different beta adrenoceptor ligands in mice with permanent occlusion of the left anterior descending coronary artery. AB - 1. We have studied the effects of three betaAR ligands (carvedilol, alprenolol, and ICI-118551) with different pharmacological profiles and negative efficacy at the beta2AR on cardiac in vivo, in vitro, biochemical and gene expression parameters in mice with permanent occlusion of the left anterior descending coronary artery. 2. Cardiac in vivo parameters were determined using Doppler studies. Mitral-wave E peak velocity (EPV) and aortic peak velocity (AoPV) decreased in the first 2 weeks postocclusion. After 3 weeks of drug treatment, EPV was improved in the carvedilol group to preocclusion values; however, a further reduction in EPV in the alprenolol and control permanent occlusion group was measured and there was no change after ICI-118551 treatment. AoPV was unchanged between weeks 2 and 5 in all groups. 3. The left atria were isolated to record isometric tension responses to isoprenaline. Permanent occlusion significantly reduced the maximum isoprenaline response to 30% of control and carvedilol increased the maximum response to isoprenaline significantly to 60%. 4. The biochemical and gene expression studies revealed different effects of the three betaAR ligands. Most notably, carvedilol reduced gene expression of myosin heavy chain beta. 5. These results indicate that chronic treatment with carvedilol is beneficial in a mouse model of myocardial damage resulting from ischaemia. We hypothesise that these beneficial effects of the drug may be because of the negative efficacy at the beta2AR, combined with beta1AR antagonism. PMID- 12721107 TI - Properties of the Na+/K+ pump current in small neurons from adult rat dorsal root ganglia. AB - 1 The present investigation was undertaken to characterize the Na(+)/K(+) pump current in small (ADP>2 MeSATP=alpha,beta-MeATP. 4. The P2 purinoceptor antagonists, suramin and pyridoxal-5-phosphate-6-azophenyl-2', 4'-disulfonic acid (PPADS), partially antagonized the ATP-induced inhibition of I(Ca), while coapplication of suramin and the P1 purinoceptor antagonist, 8-cyclopentyl-1,3-dipropylxanthine (DPCPX), virtually abolished I(Ca) inhibition. DPCPX alone partially antagonized I(Ca) inhibition. 5. Suramin antagonized the UTP-induced inhibition of I(Ca), while DPCPX had no effect. 6. Extracellular application of adenosine (ADO) also inhibited I(Ca) in a voltage-dependent manner via G(i/o)-protein activation. 7. Mainly N- and P/Q-type VDCCs were inhibited by both ATP and ADO via G(i/o) protein betagamma subunits in seemingly convergence pathways. PMID- 12721111 TI - Localization of the GSH-dependent photolabelling site of an agosterol A analog on human MRP1. AB - 1. Human multidrug resistance protein 1 (MRP1) is a 190 kDa membrane glycoprotein that confers multidrug resistance (MDR) to tumor cells. We recently demonstrated that glutathione (GSH) is required for the labelling of the C-terminal half of MRP1 with a photoanalog of agosterol A (azido AG-A). In this study, we further characterized the GSH-dependent photolabelling site of azido AG-A on MRP1. 2. An epitope-inserted MRP1, MRP1 1222HA, which has two hemagglutinin A (HA) epitopes in the extracellular loop between transmembrane segment (TM) 16 and TM17 of the transporter, could bind azido AG-A in a GSH-dependent manner. 3. Protease digestion of the photolabelled MRP1 1222HA, followed by immunoprecipitation with an anti-HA antibody suggested that the GSH-dependent azido AG-A photolabelling site on MRP1 resides in the region within TM14-17 and the cytoplasmic region proximate to the C-terminus of TM17. 4. Arg(1210) in human MRP2 that corresponds to Arg(1202) in human MRP1 has an important role in the transporting activity of MRP2. Therefore, we replaced the Arg residue at position 1202 of MRP1 with Gly. Whereas photolabelling of the mutant MRP1 R1202G was greatly reduced, it retained leukotriene C(4) (LTC(4)) transport activity and conferred Vincristine resistance in LLC-PK1 cells. 5. In summary, this study demonstrated that the GSH-dependent azido AG-A photolabelling site on MRP1 resides in the region within TM14-17 and the cytoplasmic region proximate to the C-terminus of TM17. The charged amino acid Arg(1202) proximate to TM helix 16 is of critical importance for the GSH dependent photolabelling of MRP1 with azido AG-A. Arg(1202) itself or the region nearby Arg(1202) may be involved in azido AG-A photolabelling. PMID- 12721112 TI - Different modulation by Ca2+-activated K+ channel blockers and herbimycin of acetylcholine- and flow-evoked vasodilatation in rat mesenteric small arteries. AB - 1. The present study addressed whether endothelium-dependent vasodilatation evoked by acetylcholine and flow are mediated by the same mechanisms in isolated rat mesenteric small arteries, suspended in a pressure myograph for the measurement of internal diameter. 2. In pressurized arterial segments contracted with U46619 in the presence of indomethacin, shear stress generated by the flow evoked relaxation. Thus, in endothelium-intact segments low (5.1+/-0.6 dyn cm( 2)) and high (19+/-2 dyn cm(-2)) shear stress evoked vasodilatations that were reduced by, respectively, 68+/-11 and 68+/-8% (P<0.05, n=7) by endothelial cell removal. Acetylcholine (0.01-1 microM) evoked concentration-dependent vasodilatation that was abolished by endothelial cell removal. 3. Incubation with indomethacin alone did not change acetylcholine and shear stress-evoked vasodilatation, while the combination of indomethacin with the nitric oxide (NO) synthase inhibitor, N(G),N(G)-asymmetric dimethyl-L-arginine (ADMA 1 mM), reduced low and high shear stress-evoked vasodilatation with, respectively, 52+/-15 and 58+/-10% (P<0.05, n=9), but it did not change acetylcholine-evoked vasodilatation. 4. Inhibition of Ca(2+)-activated K(+) channels with a combination of apamin (0.5 microM) and charybdotoxin (ChTX) (0.1 microM) did not change shear stress- and acetylcholine-evoked vasodilatation. In the presence of indomethacin and ADMA, the combination of apamin (0.5 microM) and ChTx (0.1 microM) increased contraction induced by U46619, but these blockers did not change the vasodilatation evoked by shear stress. In contrast, acetylcholine evoked vasodilatation was abolished by the combination of apamin and charybdotoxin. 5. In the presence of indomethacin, the tyrosine kinase inhibitor, herbimycin A (1 microM), inhibited low and high shear stress-evoked vasodilatation with, respectively, 32+/-12 and 68+/-14% (P<0.05, n=8), but it did not change vasodilatation induced by acetylcholine. In the presence of indomethacin and ADMA, herbimycin A neither changed shear stress nor acetylcholine-evoked vasodilatation. 6. The present study suggests that Ca(2+) activated K(+) channels sensitive for the combination of apamin and ChTx are involved in acetylcholine-evoked, mainly non-NO nonprostanoid factor-mediated, vasodilatation, while an Src tyrosine kinase plays a role for flow-evoked NO mediated vasodilatation in rat mesenteric small arteries. PMID- 12721113 TI - Cacospongionolide B suppresses the expression of inflammatory enzymes and tumour necrosis factor-alpha by inhibiting nuclear factor-kappa B activation. AB - 1. The marine product cacospongionolide B, a sesterterpene isolated from the Mediterranean sponge Fasciospongia cavernosa, is an inhibitor of secretory phospholipase A(2) with anti-inflammatory properties. In this work, we have studied the mechanism of action of this compound in the inflammatory response induced by zymosan in primary cells and in the mouse air pouch. 2. In mouse peritoneal macrophages, cacospongionolide B was able to downregulate the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX 2), resulting in decreased production of NO and prostaglandin E(2) (PGE(2)). This compound also reduced tumour necrosis factor-alpha (TNF-alpha) mRNA expression and TNF-alpha levels. 3. Cacospongionolide B inhibited nuclear factor-kappaB (NF kappaB)-DNA binding activity and the nuclear translocation of this transcription factor. 4. Treatment of cells with cacospongionolide B impaired NF-kappaB inhibitory protein (IkappaB-alpha) phosphorylation and enhanced IkappaB-alpha expression. 5. Inhibition of iNOS, COX-2 and inflammatory mediators was confirmed in the mouse air pouch. 6. These results show that cacospongionolide B is able to control NO, PGE(2) and TNF-alpha production in vitro and in vivo, effects likely dependent on NF-kappaB inhibition. PMID- 12721114 TI - The vasoactive peptide urotensin II stimulates spontaneous release from frog motor nerve terminals. AB - 1. The effect of urotensin II (U-II) on spontaneous transmitter release was examined in the frog to see if the biological activity of this vasoactive peptide extended to neural tissues. 2. In normal Ringer solution, frog and human U-II (fU II and hU-II, respectively) caused concentration-dependent, reversible increases in miniature endplate potential (MEPP) frequency, with hU-II about 22 times more potent than fU-II. hU-II caused a dose-dependent increase in MEPP amplitude, whereas fU-II caused an increase, followed by a decrease with higher concentrations. 3. Increasing extracellular Ca(2+) three-fold had no effect on the MEPP frequency increase to 25 microM hU-II. Pretreatment with thapsigargin to deplete endoplasmic reticulum Ca(2+) caused a 61% reduction in the MEPP frequency increase to 25 microM hU-II. 4. Pretreatment with the phospholipase C inhibitor U 73122 caused a 93% reduction in the MEPP frequency increase to 25 microM hU-II and a 15% reduction in the increase in MEPP amplitude. Pretreating with antibodies against the inositol 1,4,5-trisphosphate (IP(3)) type 1 receptor using liposomal techniques reduced the MEPP frequency increase by 83% but had no effect on MEPP amplitude. 5. Pretreating with protein kinase C inhibitors (bisindolylmaleimide I and III) had no effect on the response to 25 microM hU-II, but pretreating with protein kinase A inhibitors (H-89 and KT5720) reduced the MEPP frequency increase by 88% and completely abolished the increase in MEPP amplitude. 6. Our results show that hU-II is a potent stimulator of spontaneous transmitter release in the frog and that the effect is mediated by IP(3) and cyclic AMP/protein kinase A. PMID- 12721117 TI - Rurality and suicide. PMID- 12721115 TI - Implication of the bradykinin receptors in antigen-induced pulmonary inflammation in mice. AB - 1. The involvement of bradykinin (BK) receptors in the allergic inflammation associated with airway hyper-reactivity (AHR) was evaluated by means of the selective bradykinin B(1) receptor (BKB(1)-R) antagonists R-715 (Ac-Lys-[D betaNal(7), Ile(8)]desArg(9)-BK) and R-954 (Ac-Orn[Oic(2), alpha-MePhe(5), D betaNal(7), Ile(8)]desArg(9)-BK) or the selective bradykinin B(2) receptor (BKB(2)-R) antagonist HOE-140 (D-Arg(0)-Hyp(3)-Thi(5)-D-Tic(7)-Oic(8)-BK). Cellular migration and AHR were examined 24 h after the second ovalbumin (OA) challenge. 2. R-715 (10-500 microg kg(-1)) and R-954 (1-100 microg kg(-1)) injected intravenously (i.v.), 5 min prior to aerosol OA challenges, decreased by approximately 50% the induced lung eosinophilia in OA-sensitized mice but did not reduce AHR. 3. HOE-140 (1 microg kg(-1)) administered in the same manner, decreased mononuclear cell and eosinophil infiltration in the bronchoalveolar lavage fluid (BALF) of OA-sensitized mice. Moreover, treatment of OA-sensitized mice with HOE-140 (100 microg kg(-1)) completely abolished the AHR to carbachol. 4. The BKB(1)-R agonist desArg(9)-BK (DBK; 10-1000 microg kg(-1)) administered intratrachealy to normal mice had no effect on the basal cell counts recovered in BALF nor on the plasma extravasation, while the BKB(2)-R selective agonist BK (20 microg kg(-1)) stimulated mononuclear cell migration, neutrophilia and plasma extravasation in normal mouse lungs. Such effects were inhibited by HOE-140 (10 microg kg(-1)). 5. Our results suggest that the airway inflammatory response induced by antigen challenge in mice is mediated by stimulation of both BKB(1)-R and BKB(2)-R. PMID- 12721119 TI - Geocoding public health data. PMID- 12721120 TI - Public health and aging. PMID- 12721123 TI - Urinary tract infections and acupuncture. PMID- 12721124 TI - Reducing unhealthy behaviors: where do we start? PMID- 12721125 TI - Poverty, race, and the invisible men. PMID- 12721126 TI - Overlooked and underserved: improving the health of men of color. PMID- 12721127 TI - Improving men's health: developing a long-term strategy. PMID- 12721128 TI - Cuba's energetic AIDS doctor. PMID- 12721129 TI - Formative evaluation of a men's health center. AB - We describe an innovative approach for evaluating a men's health center. Using observation and interview, we assessed patient flow, referral patterns, patient satisfaction, and perceptions of the services' usefulness. Student assistants designed evaluation tools, hired and trained research assistants, supervised data collection, interacted with city and center officials, analyzed data, and drafted a report. To ensure patient confidentiality and anonymity, we designed an innovative observation system. The men had unique perceptions of family, requiring culturally sensitive approaches to engage them in the study. Of patients reporting to the center, 20.3% received referral services. Average satisfaction level was 5.2 (scale = 1-10). Perceived benefits to the family for 23% of respondents included cost savings, improved access, and higher service quality. PMID- 12721130 TI - Physical culture in Amherst College. Presented to the Board of Trustees of Amherst College at their annual meeting, July 8, 1869. PMID- 12721131 TI - Nathan Allen: social reformer and health advocate. PMID- 12721132 TI - The Indian club exercise. PMID- 12721133 TI - The health of men: structured inequalities and opportunities. AB - I have summarized in this article data on the magnitude of health challenges faced by men in the United States. Across a broad range of indicators, men report poorer health than women. Although men in all socioeconomic groups are doing poorly in terms of health, some especially high-risk groups include men of low socioeconomic status (SES) of all racial/ethnic backgrounds, low-SES minority men, and middle-class Black men. Multiple factors contribute to the elevated health risks of men. These include economic marginality, adverse working conditions, and gendered coping responses to stress, each of which can lead to high levels of substance use, other health-damaging behaviors, and an aversion to health-protective behaviors. The forces that adversely affect men's health are interrelated, unfold over the life course, and are amenable to change. PMID- 12721134 TI - Challenges to masculine transformation among urban low-income African American males. AB - In this article we describe and analyze the challenges faced by an intervention program that addresses the fatherhood needs of low-income urban African American males. We used life history as the primary research strategy for a qualitative evaluation of a program we refer to as the Healthy Men in Healthy Families Program to better understand the circumstances and trajectory of men's lives, including how involvement in the program might have benefited them in the pursuit of their fatherhood goals. A model of masculine transformation, developed by Whitehead, was used to interpret changes in manhood/fatherhood attitudes and behaviors that might be associated with the intervention. We combined Whitehead's model with a social ecology framework to further interpret challenges at intrapersonal, interpersonal, community, and broader societal levels. PMID- 12721135 TI - Socioeconomic status and dissatisfaction with health care among chronically ill African Americans. AB - Addressing differences in social class is critical to an examination of racial disparities in health care. Low socioeconomic status is an important determinant of access to health care. Results from a qualitative, in-depth interview study of 60 African Americans who had one or more chronic illnesses found that low-income respondents expressed much greater dissatisfaction with health care than did middle-income respondents. Low socioeconomic status has potentially deadly consequences for several reasons: its associations with other determinants of health status, its relationship to health insurance or the absence thereof, and the constraints on care at sites serving people who have low incomes. PMID- 12721136 TI - Health policy and the coloring of an American male crisis: a perspective on community-based health services. AB - Health services at the community level are organized and financed in such a way that men need access but encounter barriers to care such as poor service design, lack of insurance, and the absence of health literacy. Community health delivery systems may not be appropriate, effective, fit, or able to meet the needs they are charged to fill. Community-based health services, including health departments, are underfunded, understaffed, and unable to carry out their mission in a way that protects the health of the community. The current design for funding and delivering health care services excludes poor men, particularly men of color. Improving the health of men requires modifications in the way health care is financed, delivered, and managed. PMID- 12721137 TI - A decade of research on disparities in Medicare utilization: lessons for the health and health care of vulnerable men. AB - Medicare research has shown that there are substantial disparities by race and socioeconomic status in use of services. In this article, I review past research and discuss how findings apply specifically to vulnerable men aged 65 years or older. Six lessons from this review are identified and illustrated here. Disparities in certain measures of health are growing; to reverse this trend, substantial efforts are needed, including dissemination of information about disparities as well as testing of hypotheses regarding underlying causes. PMID- 12721138 TI - Male prisoners and HIV prevention: a call for action ignored. AB - US prison inmates are disproportionately indigent young men of color. These individuals are severely affected by HIV/AIDS, largely owing to the high-risk behavior that they engage in prior to incarceration. Researchers and practitioners have issued a call for the importance of offering HIV prevention services in prison settings. However, this call has largely been ignored. In this article, we outline reasons why these recommendations have been largely ignored, discuss innovative HIV prevention programs that are currently being implemented in prison settings, and offer recommendations for securing support for HIV prevention services in correctional settings. PMID- 12721139 TI - Slowing the epidemic of tobacco use among Asian Americans and Pacific Islanders. AB - Data on tobacco use among the Asian American/Pacific Islander (AAPI) population remain limited, although existing studies indicate that tobacco use prevalence among males from specific AAPI groups is significantly higher than in the general US male population. This high prevalence of tobacco use and the disparities in use result from social norms, targeted marketing by the tobacco industry, lack of culturally and linguistically tailored prevention and control programs, and limited impact of mainstream tobacco control programs for AAPIs. We review the available literature on tobacco use among AAPI men and women, highlight a national agenda that promotes tobacco prevention and control for AAPI communities, and acknowledge recent trends including the increase of tobacco use among AAPI women and girls. PMID- 12721141 TI - Is there a gradient in life span by position in the social hierarchy? PMID- 12721142 TI - Death of a senator: life expectancy and causes of death in 20th-century US senators. PMID- 12721140 TI - The gender gap in heart disease: lessons from Eastern Europe. AB - Why are men more susceptible to heart disease than women? Traditional risk factors cannot explain the gender gap in coronary heart disease (CHD) or the rapid increase in CHD mortality among middle-aged men in many of the newly independent states of Eastern Europe. However, Eastern European men score higher on stress-related psychosocial factors than men living in the West. Comparisons between the sexes also reveal differences in psychosocial and behavioral coronary risk factors favoring women, indicating that women's coping with stressful events may be more cardioprotective. Men's greater susceptibility to heart disease, particularly observable in many Eastern European countries, poses unique threats to public health and points to solutions in the behavioral and social arena. PMID- 12721143 TI - The health status of American Indian and Alaska native males. AB - OBJECTIVES: This study summarizes current health status information relating to American Indian and Alaska Native (AI/AN) males compared with that of AI/AN females. METHODS: I analyzed published data from the Indian Health Service for 1994 through 1996 to determine sex differences in morbidity and mortality rates and use of health care facilities. RESULTS: AI/AN males' death rates exceed those of AI/AN females for every age up to 75 years and for 6 of the 8 leading causes of death. Accidents, suicide, and homicide are epidemic among AI/AN males. Paradoxically, AI/AN males contribute only 37.9% of outpatient visits, versus 62.1% for females, and only 47% of hospitalizations excluding childbirth. CONCLUSIONS: AI/AN males suffer inordinately from a combination of increased burden of illness and lack of utilization of health care services. Programs targeted to anomie, loss of traditional male roles, and violence and alcoholism are among the most urgently needed. PMID- 12721145 TI - Receipt of preventive care among adults: insurance status and usual source of care. AB - OBJECTIVES: This study ascertained the separate and combined effects of having insurance and a usual source of care on receiving preventive services. METHODS: Descriptive and multivariate analyses of 1996 Medical Expenditure Panel Survey data were conducted. RESULTS: Receipt of preventive services was strongly associated with insurance and a usual source of care. Significant differences were found between insured adults with a usual source of care, who were most likely to have received services, compared with uninsured adults without regular care, who were least likely to have received services. Those with either a usual source of care or insurance had intermediate levels of preventive services. CONCLUSIONS: Having a usual source of care and health insurance are both important to achieving national prevention goals. PMID- 12721144 TI - Informed consent for cancer screening with prostate-specific antigen: how well are men getting the message? AB - OBJECTIVES: This study examined knowledge about prostate-specific antigen (PSA) screening among African Americans and Whites. Because PSA screening for prostate cancer is controversial, professional organizations recommend informed consent for screening. METHODS: Men (n = 304) attending outpatient clinics were surveyed for their knowledge about and experience with screening. RESULTS: Most men did not know the key facts about screening with PSA. African Americans appeared less knowledgeable than Whites, but these differences were mediated by differences in educational level and experience with prostate cancer screening. CONCLUSIONS: Public health efforts to improve informed consent for prostate cancer screening should focus on highlighting the key facts and developing different approaches for men at different levels of formal education and prior experience with screening. PMID- 12721146 TI - Racial/ethnic disparities in the use of mental health services in poverty areas. AB - OBJECTIVES: This study examined racial/ethnic disparities in mental health service access and use at different poverty levels. METHODS: We compared demographic and clinical characteristics and service use patterns of Whites, Blacks, Hispanics, and Asians living in low-poverty and high-poverty areas. Logistic regression models were used to assess service use patterns of minority racial/ethnic groups compared with Whites in different poverty areas. RESULTS: Residence in a poverty neighborhood moderates the relationship between race/ethnicity and mental health service access and use. Disparities in using emergency and inpatient services and having coercive referrals were more evident in low-poverty than in high-poverty areas. CONCLUSIONS: Neighborhood poverty is a key to understanding racial/ethnic disparities in the use of mental health services. PMID- 12721147 TI - Markers of access to and quality of primary care for aboriginal people in Ontario, Canada. AB - OBJECTIVES: We evaluated primary care accessibility and quality for Ontario's aboriginal population. METHODS: We compared a defined aboriginal cohort with nonaboriginal populations with analogous geographic isolation and low socioeconomic status. We determined rates of hospitalization for the following indicators of adequacy of primary care: ambulatory care-sensitive (ACS) conditions and utilization of referral care-sensitive (RCS) procedures from administrative databases. RESULTS: ACS hospitalization rates, relative to the general population, were 2.54, 1.50, and 1.14 for the aboriginal population, the geographic control populations, and the socioeconomic control populations, respectively. The relative RCS procedure utilization rates were 0.64, 0.91, and 1.00, respectively. CONCLUSIONS: The increased ACS hospitalization rate and reduced RCS procedure utilization rate suggest that northern Ontario's aboriginal residents have insufficient or ineffective primary care. PMID- 12721148 TI - Addressing urban health in Detroit, New York City, and Seattle through community based participatory research partnerships. AB - OBJECTIVE: This study describes key activities integral to the development of 3 community-based participatory research (CBPR) partnerships. METHODS: We compared findings from individual case studies conducted at 3 urban research centers (URCs) to identify crosscutting adaptations of a CBPR approach in the first 4 years of the partnerships' development. RESULTS: Activities critical in partnership development include sharing decision-making, defining principles of collaboration, establishing research priorities, and securing funding. Intermediate outcomes were sustained CBPR partnerships, trust within the partnerships, public health research programs, and increased capacity to conduct CBPR. Challenges included the time needed for meaningful collaboration, concerns regarding sustainable funding, and issues related to institutional racism. CONCLUSIONS: The URC experiences suggest that CBPR can be successfully implemented in diverse settings. PMID- 12721149 TI - Migration and HIV risk behaviors: Puerto Rican drug injectors in New York City and Puerto Rico. AB - OBJECTIVES: We compared injection-related HIV risk behaviors of Puerto Rican current injection drug users (IDUs) living in New York City and in Puerto Rico who also had injected in the other location with those who had not. METHODS: We recruited Puerto Rican IDUs in New York City (n = 561) and in Puerto Rico (n = 312). Of the former, 39% were "newcomers," having previously injected in Puerto Rico; of the latter, 14% were "returnees," having previously injected in New York. We compared risk behaviors within each sample between those with and without experience injecting in the other location. RESULTS: Newcomers reported higher levels of risk behaviors than other New York IDUs. Newcomer status (adjusted odds ratio [OR] = 1.62) and homelessness (adjusted OR = 2.52) were significant predictors of "shooting gallery" use; newcomer status also predicted paraphernalia sharing (adjusted OR = 1.67). Returnee status was not related to these variables. CONCLUSIONS: Intervention services are needed that target mobile populations who are coming from an environment of high-risk behavior to one of low-risk behavior. PMID- 12721150 TI - Leisure time physical activity and disease-specific mortality among men with chronic bronchitis: evidence from the Whitehall study. AB - OBJECTIVES: This study examined the association between leisure time physical activity and cause-specific mortality among male Whitehall Study participants with chronic bronchitis. METHODS: Rate ratios were calculated for 4 mortality outcomes, according to level of activity and baseline bronchitis status, in a 25 year follow-up of 6479 men. RESULTS: After multiple adjustment for potential confounding or mediating variables, activity was inversely related to all-cause, cardiovascular, coronary heart disease, and noncardiovascular mortality among men free of chronic bronchitis. Among men with bronchitis, weak, nonsignificant positive associations were observed between activity and these outcomes, with the exception of noncardiovascular mortality. CONCLUSIONS: The suggestion of a positive activity-mortality association among individuals with chronic bronchitis albeit weak and nonsignificant-requires further investigation. PMID- 12721151 TI - R.J. Reynolds' targeting of African Americans: 1988-2000. AB - OBJECTIVES: The purpose of this study was to describe RJ Reynolds (RJR) Tobacco Company's strategy for targeting African Americans, as revealed in tobacco industry documents and magazine advertisements. METHODS: The authors searched industry documents to determine RJR's strategies and analyzed magazine advertising during 2 periods: the time of the launch of the company's Uptown cigarette (1989-1990) and a decade later (1999-2000). RESULTS: RJR's efforts to target the African American market segment existed before and after Uptown, and the company's strategy was largely implemented via other RJR brands. Advertisements featured mentholated cigarettes, fantasy/escape, expensive objects, and nightlife. CONCLUSIONS: To help all populations become tobacco-free, tobacco control practitioners must understand and counter tobacco industry strategies. PMID- 12721152 TI - African Americans' attitudes toward cigarette excise taxes. AB - OBJECTIVES: This study examined African Americans' opinions regarding cigarette excise taxes and other tobacco control issues. METHODS: A stratified cluster sample of US congressional districts represented by African Americans was selected. African Americans from 10 districts were interviewed. RESULTS: Forty seven percent of respondents stated that taxes on tobacco products should be increased, whereas about 30% believed that they should be reduced. Almost 75% disagreed that raising taxes on tobacco products is unfair to African Americans, and 57.9% reported that they would not be opposed to increasing taxes on cigarettes even if low-income smokers would be hit the hardest. CONCLUSIONS: The present results indicate substantial support for cigarette excise taxes among African Americans. PMID- 12721153 TI - Glucocorticoid-induced skeletal muscle atrophy is associated with upregulation of myostatin gene expression. AB - The mechanisms by which excessive glucocorticoids cause muscular atrophy remain unclear. We previously demonstrated that dexamethasone increases the expression of myostatin, a negative regulator of skeletal muscle mass, in vitro. In the present study, we tested the hypothesis that dexamethasone-induced muscle loss is associated with increased myostatin expression in vivo. Daily administration (60, 600, 1,200 micro g/kg body wt) of dexamethasone for 5 days resulted in rapid, dose-dependent loss of body weight (-4.0, -13.4, -17.2%, respectively, P < 0.05 for each comparison), and muscle atrophy (6.3, 15.0, 16.6% below controls, respectively). These changes were associated with dose-dependent, marked induction of intramuscular myostatin mRNA (66.3, 450, 527.6% increase above controls, P < 0.05 for each comparison) and protein expression (0.0, 260.5, 318.4% increase above controls, P < 0.05). We found that the effect of dexamethasone on body weight and muscle loss and upregulation of intramuscular myostatin expression was time dependent. When dexamethasone treatment (600 micro g. kg-1. day-1) was extended from 5 to 10 days, the rate of body weight loss was markedly reduced to approximately 2% within this extended period. The concentrations of intramuscular myosin heavy chain type II in dexamethasone treated rats were significantly lower (-43% after 5-day treatment, -14% after 10 day treatment) than their respective corresponding controls. The intramuscular myostatin concentration in rats treated with dexamethasone for 10 days returned to basal level. Concurrent treatment with RU-486 blocked dexamethasone-induced myostatin expression and significantly attenuated body loss and muscle atrophy. We propose that dexamethasone-induced muscle loss is mediated, at least in part, by the upregulation of myostatin expression through a glucocorticoid receptor mediated pathway. PMID- 12721154 TI - Glucose-dependent insulinotropic peptide stimulates thymidine incorporation in endothelial cells: role of endothelin-1. AB - We have previously characterized the receptor for glucose-dependent insulinotropic polypeptide (GIPR) in vascular endothelial cells (EC). Different EC types were found to contain distinct GIPR splice variants. To determine whether activation of the GIPR splice variants resulted in different cellular responses, we examined GIP effects on human umbilical vein endothelial cells (HUVEC), which contain two GIPR splice variants, and compared them with a spontaneously transformed human umbilical vein EC line, ECV 304, which contains four GIPR splice variants. GIP dose-dependently stimulated HUVEC and ECV 304 proliferation as measured by [3H]thymidine incorporation. GIP increased endothelin-1 (ET-1) secretion from HUVEC but not from ECV 304. Use of the endothelin B receptor blocker BQ-788 resulted in an inhibition of [3H]thymidine incorporation in HUVEC but not in ECV 304. These findings suggest that, although GIP increases [3H]thymidine incorporation in both HUVEC and ECV 304, this proliferative response is mediated by ET-1 only in HUVEC. These differences in cellular response to GIP may be related to differences in activation of GIPR splice variants. PMID- 12721155 TI - NO production by cNOS and iNOS reflects blood pressure changes in LPS-challenged mice. AB - Increased nitric oxide (NO) production is the cause of hypotension and shock during sepsis. In the present experiments, we have measured the contribution of endothelial (e) and inducible (i) nitric oxide synthase (NOS) to systemic NO production in mice under baseline conditions and upon LPS treatment (100 microg/10 g ip LPS). NO synthesis was measured by the rate of conversion of l [guanidino-15N2]arginine to l-[ureido-15N]citrulline, and the contribution of the specific NOS isoforms was evaluated by comparing NO production in eNOS-deficient [(-/-)] and iNOS(-/-) mice with that in wild-type (WT) mice. Under baseline conditions, NO production was similar in WT and iNOS(-/-) mice but lower in eNOS( /-) mice [WT: 1.2 +/- 0.2; iNOS(-/-): 1.2 +/- 0.2; eNOS(-/-): 0.6 +/- 0.3 nmol. 10 g body wt-1. min-1]. In response to the challenge with LPS (5 h), systemic NO production increased in WT and eNOS(-/-) mice but fell in iNOS(-/-) mice [WT: 2.7 +/- 0.3; eNOS(-/-): 2.2 +/- 0.6; iNOS(-/-): 0.7 +/- 0.1 nmol. 10 g body wt-1. min 1]. After 5 h of LPS treatment, blood pressure had dropped 14 mmHg in WT but not in iNOS(-/-) mice. The present findings provide firm evidence that, upon treatment with bacterial LPS, the increase of NO production is solely dependent on iNOS, whereas that mediated by cNOS is reduced. Furthermore, the data show that the LPS-induced blood pressure response is dependent on iNOS. PMID- 12721156 TI - Characterization of leptin pulse dynamics and relationship to fat mass, growth hormone, cortisol, and insulin. AB - To investigate the regulation of leptin secretion and pulsatility by fat mass, we performed overnight leptin sampling every 20 min for 12 h and compared leptin dynamics with total body and regional fat measurements in 20 healthy male subjects. Simultaneous growth hormone (GH), cortisol, and insulin levels were assessed to determine relatedness and synchronicity during overnight fasting. Deconvolution analyses were performed to determine simultaneous hormonal dynamics, synchronicity, and interrelatedness using cross-correlation and cross approximate entropy (X-ApEn) analyses. Subjects demonstrated 4.7 +/- 0.4 leptin pulses/12 h. Leptin secretion correlated highly with total body fat (r = 0.78, P < 0.001) and regional fat depots. In contrast, leptin pulsatility did not correlate with total fat (r = 0.07, P = 0.785) or other measures of fat. There was synchronicity between GH and leptin (lag -39 minutes), cortisol and leptin (lag -211 min), and leptin and insulin, with leptin following insulin by 275 min. The mean random X-ApEn was significant between leptin and GH (0.854 +/- 0.030), cortisol (0.891 +/- 0.023), and insulin (0.868 +/- 0.034), demonstrating a high degree of regularity and pattern frequency. These data demonstrate differential regulation of leptin secretion and pulsatility in adipocytes and suggest that the leptin pulse generator is extrinsic to fat, whereas fat mass acts as an amplifier to modulate secretion and amplitude for a given pulsatility. We demonstrate synchronicity between leptin and GH, cortisol, and insulin. The directionality of the cross correlation suggests a temporal construct in which changes in leptin follow those of insulin but precede those of GH and cortisol during overnight fasting. PMID- 12721157 TI - Expression of uncoupling protein 3 is upregulated in skeletal muscle during sepsis. AB - Uncoupling protein 3 (UCP3) is a member of the mitochondrial transporter superfamily that is expressed primarily in skeletal muscle. UCP3 is upregulated in various conditions characterized by skeletal muscle atrophy, including hyperthyroidism, fasting, denervation, diabetes, cancer, lipopolysaccharide (LPS), and treatment with glucocorticoids (GCs). The influence of sepsis, another condition characterized by muscle cachexia, on UCP3 expression and activity is not known. We examined UCP3 gene and protein expression in skeletal muscles from rats after cecal ligation and puncture and from sham-operated control rats. Sepsis resulted in a two- to threefold increase in both mRNA and protein levels of UCP3 in skeletal muscle. Treatment of rats with the glucocorticoid receptor antagonist RU-38486 prevented the sepsis-induced increase in gene and protein expression of UCP3. The UCP3 mRNA and protein levels were increased 2.4- to 3.6 fold when incubated muscles from normal rats were treated with dexamethasone (DEX) and/or free fatty acids (FFA) ex vivo. In addition, UCP3 mRNA and protein levels were significantly increased in normal rat muscles in vivo with treatment of either DEX or FFA. The results suggest that sepsis upregulates the gene and protein expression of UCP3 in skeletal muscle, which may at least in part be mediated by GCs and FFA. PMID- 12721158 TI - The design, publication and interpretation of research in Subfertility Medicine: uncomfortable issues and challenges to be faced. PMID- 12721159 TI - Stimulated intra-uterine insemination is not a natural choice for the treatment of unexplained subfertility. Should the guidelines be changed? AB - The Royal College of Obstetricians and Gynaecologists Guidelines for the management of infertility in secondary care (1998) states that stimulated intrauterine insemination (IUI) has Grade A evidence to confirm its effective use in unexplained infertility. This paper challenges that assertion after closer assessment of the papers upon which it was based. With the current appraisal of the Guidelines as part of the 'scope' of the assessment of infertility management by the National Institute of Clinical Excellence it is important that the guidelines are indeed critically reviewed prior to their updated publication. With the emphasis on minimizing the risk of multiple pregnancies, a formal trial of stimulated IUI is called for in order to justify its continued use in this setting. Strict attention to limiting the number of follicles treated in verified unexplained subfertility is necessary to validate such a trial. PMID- 12721160 TI - Stimulated intra-uterine insemination is not a natural choice for the treatment of unexplained subfertility. Current best evidence for the advanced treatment of unexplained subfertility. AB - When conventional treatment of unexplained infertility has not been successful, the next choice lies between either gonadotrophin treatment with intrauterine insemination (FSH/IUI) and IVF. Cohort studies indicate that FSH/IUI is less effective while the cost per birth is higher with IVF. The relevant evidence should come from randomized controlled trials, but the results need to be recalculated for the outcome of greatest interest, a singleton live birth. Judged by this standard, FSH/IUI is statistically superior to no treatment, but the effect is small in couples with unexplained infertility. By the same standard, IVF is superior to FSH/IUI treatment, but this benefit is achieved only at considerable cost. The evidence is not robust, comprising only a few trials which involve a minute fraction of the existing patients with unexplained infertility. Current best evidence is consistent with a progression from low-tech to high-tech treatment, but it is not convincing enough to support a rigid management protocol. A large multicentre factorial trial is needed to evaluate the relative value of existing empiric treatments for unexplained infertility. PMID- 12721161 TI - Stimulated intra-uterine insemination is not a natural choice for the treatment of unexplained subfertility. 'Effective treatment' or 'not a natural choice'? AB - Helping couples to choose appropriate therapy for their unexplained subfertility demands a review of evidence for treatment benefit and harm, in the context of both patients' and clinicians' experience and perspective. Ovulation induction (OI) with clomiphene (CC) or gonadotrophin (FSH) and intrauterine insemination (IUI) are often chosen before resorting to IVF. The appropriateness of starting with these low and intermediate intensity treatments is supported by evidence that CC/IUI increases cycle fecundity two- to three-fold, and FSH/IUI, three- to five-fold over the baseline chance of pregnancy in this patient group. While both OI/IUI and IVF have adverse effects which deserve vigorous attention, particularly multiple pregnancy and ovarian hyperstimulation syndrome (OHSS), the balance between benefits and costs often favours OI/IUI. The cost per live birth and potential for OHSS appears lower with OI/IUI, and the proportion of multiple pregnancies similar to that seen with IVF. For these as well as physical and spiritual reasons, OI/IUI is often a natural starting point for couples, especially when female age and duration of subfertility are favourable. PMID- 12721162 TI - Immune infertility: towards a better understanding of sperm (auto)-immunity. The value of proteomic analysis. AB - Antisperm antibodies (ASA) in the male cause an autoimmune disease 'immune infertility'. It has to be clarified whether each antibody binding to an antigen, which is identified on the sperm surface, also influences sperm function. In the past, the clinical interest in ASA was hampered by the fact that a standardized assay for the detection of ASA was lacking. There are several methods to characterize the cognate antigens of ASA. In the following article, reports from the recent literature of immunologically characterized sperm proteins-as cognate antigens of naturally occurring ASA or of artificially produced antibodies-will be quoted with respect to different sperm functions. As a practical consequence of the research on ASA-related sperm proteomics, those ASA that decrease male fertility by inhibiting sperm functions essential for fertilization will be identified. PMID- 12721163 TI - Safety issues in assisted reproduction technology. From theory to reality--just what are the data telling us about ICSI offspring health and future fertility and should we be concerned? AB - Concerns about the effects of ICSI on offspring health and fertility include the rate of chromosomal anomalies, cystic fibrosis (CF) gene mutations associated with congenital bilateral absence of the vas deferens (CBAVD) and Y-chromosome microdeletions. The evidence in favour of screening for these in the presence of azoospermia or severe oligozoospermia is beyond debate. Concerns requiring further investigation include the effects of ICSI on imprinted genes and genes involved in DNA replication error repair. There is evidence of an increased risk of low birth weight, cerebral palsy and major birth defects following assisted reproductive technologies (ART), including ICSI, although the causes remain unknown. Given the large studies required to investigate these questions, particularly for rarer genetic conditions, we may simply have to accept that we may not know for many years if there are increased risks associated with ICSI. It would be prudent, however, to acknowledge this as a possibility and counsel patients accordingly. In terms of certainty of outcome and magnitude of impact the single most important health effect of ART for the offspring remains the iatrogenic multiple pregnancy rate. A reduction of iatrogenic multiples is the single most important and achievable means of preventing cerebral palsy currently available. Once achieved, the occurrence of rare genetic conditions will assume greater importance. PMID- 12721164 TI - Intrafamilial sperm donation: ethical questions and concerns. AB - Sperm donation seems to be the only solution in cases of negative results with testicular sperm extraction (TESE). However, it implies a genetic dissociation between husband and offspring that results in psychological stress, as well as ethical and existential dilemmas for the couple. Facing these dilemmas, some couples prefer father-to-son donation as an alternative solution. However, father to-son sperm donation involves many potential medical, ethical and emotional risks. A broad discussion within the scientific community on these issues should contribute towards a better approach of these cases. PMID- 12721165 TI - Coasting-what is the best formula? AB - Coasting is a method to decrease the incidence of ovarian hyperstimulation syndrome (OHSS), which involves withdrawing exogenous gonadotrophins until the serum estradiol (E(2)) level decreases. The application of this strategy, as it appears in the literature, has been variable, with heterogeneous criteria for initiating and ending the coasting process and as a result, reports of efficacy are inconsistent. In attempt to establish a recommended protocol for coasting we reviewed and analysed 10 relevant studies, found by a Medline search. Based on the data collected, coasting should be initiated when the serum E(2) concentration exceeds 3000 pg/ml, but not unless the leading follicles reach a diameter of 15-18 mm. Its duration should be limited to <4 days, thus, preventing the decrease in implantation and pregnancy rates that occur after longer periods of coasting. Administration of hCG should be withheld until serum E(2) falls below 3000 pg/ml. Based on the published data, these suggested guidelines result in an acceptably low incidence of severe OHSS (<2%) and provide satisfactory fertilization and pregnancy rates (55-71% and 36.5-63% respectively). A multicentre randomized prospective study would help to confirm the effectiveness of this approach. PMID- 12721166 TI - Single versus double intrauterine insemination in stimulated cycles for subfertile couples: a systematic review based on a Cochrane review. AB - This paper is based on a Cochrane review published in The Cochrane Library, issue 1, 2003 (see www.update- software.com) with permission from The Cochrane Collaboration and Update Software. Cochrane reviews are regularly updated as new information becomes available and in response to comments and criticisms, and The Cochrane Library should be consulted for the most recent version of the review. BACKGROUND: The objective of this review was to determine, from the best available evidence, the difference in outcome using single versus double intrauterine insemination (IUI) in stimulated cycles for subfertile couples. METHODS: The principles of the Cochrane Menstrual Disorders and Subfertility Group were employed. Randomized controlled trials with a parallel design, comparing single versus double IUI in subfertile couples, would be eligible. The main outcome measures included live birth rate and pregnancy rate per couple (and per cycle). RESULTS: Three studies involving 386 women were included. The results of pregnancy rate per couple, of two studies showed no significant effect of using double insemination [Peto odds ratio (OR) 1.45; 95% confidence interval (CI) 0.78-2.70]. The results of pregnancy rate per cycle of the included studies favoured double insemination, however this is not an eligible outcome measure. CONCLUSIONS: Based on the results of two trials, double intrauterine insemination showed no significant benefit over single IUI in the treatment of subfertile couples with partner semen. There are no meaningful data to offer advice on the basis of this review. A randomized controlled trial of single versus double IUI is justified. PMID- 12721167 TI - Gaps in the evidence for fertility treatment-an analysis of the Cochrane Menstrual Disorders and Subfertility Group database. AB - BACKGROUND: The randomized controlled trial is considered the best approach to assess the effectiveness of treatments. The aim was to summarize the available evidence and determine gaps in the evidence for clinical decision making in subfertility. METHODS: A search of the Cochrane Library for Menstrual Disorders and Subfertility Group reviews was undertaken and, where the reviews were related to subfertility, the authors' conclusions were appraised and correlated with the results and meta-analysis sections of the reviews. Each review was then categorized as to what extent it had answered the clinical question posed by the reviewers. RESULTS: Of 38 subfertility reviews currently or previously published on the Cochrane Library from the Menstrual Disorders and Subfertility Group, 12 reviews concluded that there was evidence of effectiveness of the interventions studied. There was insufficient evidence of effectiveness in 26 reviews, from which the authors of 23 reviews called for further research. A tabulated summary of the review conclusions is presented. CONCLUSION: Cochrane subfertility reviews have eliminated some gaps in the evidence and highlighted others. Future clinical trial design should focus on adequate power and reporting the major outcome of live-births per couple as well as adverse events. PMID- 12721168 TI - A study of teratogenicity of hydrosalpinx fluid using a whole rat embryo culture model. AB - BACKGROUND: Hydrosalpinx fluid may be toxic to sperm and early embryo growth. Information concerning the effect of hydrosalpinx fluid on embryo development during organogenesis is lacking. METHODS: Rat embryos at gestational day 9.5 were cultured for 48 h with 80% rat serum and 20% of either hydrosalpinx fluid (study group) or lactated Ringer's solution (control group). Embryos were scored for growth and development at the end of the culture period. RESULTS: Hydrosalpinx fluid, collected from 10 patients, was tested for embryotoxicity individually. Median total morphological scores were significantly lower in embryos exposed to hydrosalpinx fluid from three of the 10 patients (43.0 versus 47.0, P = 0.01; 36.0 versus 45.0, P < 0.001; 36.0 versus 46.5, P = 0.003). This was accompanied by a significant reduction in median yolk sac diameter (4.0 versus 5.2 mm, P < 0.001 and 4.0 versus 5.0 mm, P < 0.001) and somite number (17.5 versus 22.5, P < 0.001 and 17.0 versus 21.5, P = 0.008) in the latter two patients. CONCLUSIONS: Hydrosalpinx fluid in some patients may contain toxin(s) that is potentially teratogenic. PMID- 12721169 TI - Nitric oxide inhibits polyamine-induced apoptosis in the human extravillous trophoblast cell line SGHPL-4. AB - BACKGROUND: Polyamines are regulators of proliferation and differentiation in mammalian cells. They are also known to regulate cell survival and apoptosis, although their precise function varies between cell types. We have investigated the effect of polyamines on the apoptosis of human extravillous trophoblasts. METHODS: Using the extravillous trophoblast-derived cell line SGHPL-4 we performed time-lapse microscopy studies to evaluate the induction of apoptosis following exposure to polyamines. RESULTS: The polyamines spermine, and to a lesser extent spermidine, were able to induce apoptosis in extravillous trophoblasts. The induction of apoptosis occurred rapidly and was accompanied by DNA fragmentation and morphological changes consistent with the onset of apoptosis. Apoptosis was inhibited by the broad-spectrum caspase inhibitor Z-VAD fmk, although no activity was detected using assays for caspase-2, -3, -6, -8 or 9 activity. We demonstrated that an oxidation product of spermine accounted for the induction of apoptosis and implicated the formation of hydrogen peroxide as this oxidation product. We have also demonstrated that exposure to nitric oxide inhibited the onset of spermine-induced apoptosis. CONCLUSIONS: Spermine and spermidine induce apoptosis in extravillous trophoblasts following their oxidation and the production of hydrogen peroxide. Nitric oxide is able to inhibit this apoptosis. PMID- 12721170 TI - Immunocompetent cells in the endometrium of fetuses and children. AB - BACKGROUND: Although the immunocompetent cells of the adult human endometrium are well characterized, there is little information about these cells in the developing uterus. This study was undertaken to investigate the distribution of leukocyte subpopulations in the endometrium of fetuses and children. METHODS: Uterine tissue obtained at autopsy from fetuses (n = 11) and neonates/children (n = 9) between 17 weeks gestation and 5(1/2) years of age was investigated with antibodies against various leukocyte subsets by immunohistochemical staining techniques. RESULTS: The densities of CD45+ and CD68+ cells were significantly higher in the endometrium of neonates/children than in that of fetuses. CD14+ monocytes represented the largest leukocyte subpopulation in both groups. CD56+ natural killer cells and HLA-DR+ antigen-presenting cells were absent from fetal endometrium. There were no differences in density of CD3+ T cells between the two groups, but CD4+ T helper cells were found only in fetal endometrium. CONCLUSIONS: The endometrial leukocyte population of fetuses and small children is different from that seen in adult women. The appearance of CD56+ and HLA-DR+ cells in endometrium seems to be a post-natal event, which may be induced by the changes in hormone levels and/or the adaptation of the local immune system to the changing microenvironment. PMID- 12721171 TI - In-utero androgen exposure and 2nd to 4th digit length ratio-comparisons between healthy controls and females with classical congenital adrenal hyperplasia. AB - BACKGROUND: Soft tissue measurements from the hand reveal lower second to fourth finger digit ratios (2D:4D) in males compared with females. The relatively longer 4th finger observed in males may be related to increased fetal exposure to androgens influencing the expression of Hox genes. METHODS: We have measured 2D:4D ratios in 69 healthy females [median age 9.3 (range 1.9-17) years], 77 control males [median age 13.86 (2.1-20.3) years] and in 66 females with classical virilizing congenital adrenal hyperplasia (CAH) (median age 8.5 (1.1 16.2) years] who are known to be exposed to high concentrations of androgens in utero. Measurements were determined from X-rays of the left hand using vernier callipers. Intra-observer variability in measurement technique was 0.01%. RESULTS: Control males had a significantly lower mean (SD) 2D:4D ratio [0.918 (0.029)] compared with female patients [0.927 (0.029), ANOVA P = 0.02]. No difference in 2D:4D ratio was observed between CAH females [0.925 (0.021)] and control females [0.927 (0.029)]. In contrast, 2D:4D ratio in males were significantly lower compared with CAH females (P = 0.03). CONCLUSIONS: 2D:4D ratios determined directly from radiographs of the left hand confirm significant differences between males and females. However, female patients with virilizing CAH do not have a male digit ratio pattern suggesting that in the left hand digit ratio development is not influenced by in-utero exposure to androgens. PMID- 12721172 TI - Cardiovascular risk factors in young Czech females with polycystic ovary syndrome. AB - BACKGROUND: Polycystic ovary syndrome (PCOS) could be associated with a variety of signs of metabolic syndrome. The aim of our study was to compare the cardiovascular risk factors in PCOS women and in a control group selected from a random population sample. METHODS AND RESULTS: 50 PCOS women with a mean (+/-SD) age of 30.7 +/- 4.2 years, and 335 controls with a mean age of 29.9 +/- 3.1 years selected from a random population sample of nine districts of the Czech Republic were compared for basic anthropometric characteristics, blood pressure, plasma lipids and fasting glucose. PCOS women had a significantly higher body mass index (BMI). After adjusting for BMI, PCOS women had higher blood pressure and LDL cholesterol, and lower HDL and HDL-cholesterol/total ratio. Arterial hypertension was more prevalent in PCOS women than in controls. There was no difference in the prevalence of impaired fasting glucose between both groups. Impaired glucose tolerance was found in 11.8% of PCOS women. Diabetes mellitus was more frequent in PCOS families. CONCLUSIONS: Czech PCOS women, even in their thirties, show a significantly worse cardiovascular risk profile than a control group selected from a random population sample. The differences cannot be explained by obesity. PMID- 12721173 TI - Associations between patients with endometriosis and HLA class II; the analysis of HLA-DQB1 and HLA-DPB1 genotypes. AB - BACKGROUND: Although the aetiology of endometriosis remains unclear, many immunological abnormalities involving changes in cell-mediated and humoral immunity may be associated with endometriosis. Several disorders are thought to be associated with particular HLA antigen types. This study examines the possible association between HLA-DQ and DP. METHODS: A total of 83 patients diagnosed with endometriosis following laparoscopic examination were typed for the HLA-DQB1 and DPB1 alleles using PCR-restriction fragment length polymorphism (RFLP). The HLA DQB1 and DPB1 allele frequencies in these patients and in 222 controls were compared. RESULTS: The prevalence of HLA-DQB1*0301 in the patient group was 16.3% (27/166 alleles), compared with 8.3% in the overall control group (37/444 alleles) and 7.7% in the females of the control group (18/234 alleles). Thus, the prevalence of the HLA-DQB1*0301 allele was significantly greater in patients with endometriosis, compared with the general controls [OR 2.13, 95% CI 1.25-3.64, P = 0.004 (chi(2) analysis), Corrected P-values; Pc = 0.049] and with the general female controls [OR 2.33, 95% CI 1.24-4.39, P = 0.008 (chi(2) analysis), Pc; NS]. There was no significant association in the frequencies of DPB1 alleles between the patients and controls. CONCLUSIONS: The HLA systems may be involved in the aetiology of endometriosis, although further study is needed. PMID- 12721174 TI - Intraperitoneal levonorgestrel-releasing intrauterine device following uterine perforation: the role of progestins in adhesion formation. AB - BACKGROUND: Intrauterine contraception is a widely used, highly effective means of birth control. Uterine perforation is a serious, albeit rare, complication of intrauterine device (IUD) use. Although uterine perforation by levonorgestrel releasing (20 micro g/day) intrauterine system (LNG-IUS) has already been reported, the peritoneal adhesion potential of this IUD is unknown. METHODS: The medical files of all patients diagnosed with an intra-peritoneal IUD between the years 1990-2002 at Hadassah Medical Center were reviewed. Histopathological study of peritoneal adhesion tissue adjacent to levonorgestrel medicated IUD was conducted in one case. RESULTS: Eight cases of dislocated IUDs were found. Four cases used LNG-IUS and four other cases used copper-IUD. Laparoscopy for IUD removal disclosed mild local peritoneal adhesions between omentum and pelvic organs in all cases. No difference was noted in the appearance of the peritoneum in the presence of either a copper-IUD or LNG-IUS. Histological examination of peritoneal tissue encasing the levonorgestrel-intrauterine system revealed loose connective tissue with aggregates of submesothelial cells with a pseudo-decidual change. Immunohistochemical staining for progesterone receptor was negative. CONCLUSIONS: The peritoneal adhesions potential of LNG-IUS is low, similar to that of the copper-bearing IUD. PMID- 12721175 TI - Contraception: from accessibility to efficiency. AB - BACKGROUND: Despite the widespread use of medical contraception in France, the prevalence of abortion remains stable. A cross-sectional population-based survey was designed to study the characteristics of current contraceptive use in France, the different types of contraceptive failure, and the reasons reported for not using contraception. METHODS: A representative sample of 14 704 French households was randomly selected from the telephone directory. All women who in the last 5 years had had an abortion or whose last pregnancy was unintended were selected (n = 1034), whilst a fraction (n = 1829) of the other women were randomly selected. Altogether, 2863 women answered the questionnaire. RESULTS: Only 2.7% of the women aged 18-44 years who did not wish to conceive and were sexually active did not use any contraception. However, 33% of pregnancies were estimated to be unplanned, of which 50% ended in an abortion. A total of 65% of the reported unplanned pregnancies occurred among women using contraception (21% the pill, 9% IUD, 12% condoms, 23% other method). The main reasons given for these contraceptive failures were the misuse of the methods or the failure of the partner to withdraw. Reasons for not using any contraception go beyond a simple lack of information. CONCLUSIONS: The importance of-and the reasons mentioned for contraceptive failure show a misadaptation between women's contraceptive needs and the method they use. Improving the training of family planning providers remains a major goal to ensure that women use a contraceptive method that fits their social and sexual lifestyle. PMID- 12721176 TI - Common statistical errors in the design and analysis of subfertility trials. AB - BACKGROUND: The quality of clinical trials has received increasing attention with the growth of evidence-based medicine and systematic reviews. We aimed to identify whether errors and omissions commonly encountered when undertaking Cochrane reviews in this field are still passing peer review. METHODS: We undertook a review of trials published in 2001 by two major journals. We selected from Medline only trials in which authors compared pregnancy rates under two interventions by allocating women to different groups. RESULTS: We identified 39 trials meeting our criteria. Six trials were fatally flawed by design, either by inappropriate use of a cross-over design or by systematic allocation described by the authors as 'random'. Only six reports claimed to apply the intention-to-treat principle, and the principle was misunderstood by four of these. Only five trials reported live birth rates sufficiently to allow valid meta-analysis. Most trials (82%) included at least one 'unit of analysis' error. CONCLUSIONS: We selected simple trials from respected journals, assuming that our sample would represent trials of highest methodological quality in the field. Nevertheless, the standards of design, analysis and reporting of many subfertility trials are not sufficient to allow reliable interpretation of results, or inclusion in meta analyses. PMID- 12721177 TI - Pitfalls in the design and analysis of efficacy trials in subfertility. PMID- 12721178 TI - Participation of the sperm proteasome in human fertilization. AB - BACKGROUND: Fertilization in mammals comprises the sequential interactions of the sperm with the cumulus oophorus, zona pellucida, and oocyte plasma membrane. Here we investigate proteasome activity in human sperm and its possible involvement during the fertilization process. METHODS: Proteasome activity was measured in intact sperm and in sperm extracts using the fluorogenic substrate Suc-Leu-Leu Val-Tyr-AMC, in the presence or absence of the specific proteasome inhibitor, clasto-lactacystin beta-lactone. The participation of the proteasome was evaluated during (i) sperm-zona binding using the hemizona assay; (ii) zona pellucida-induced acrosome reactions with a pulse and chase design; (iii) progesterone-induced acrosome reactions incubating overnight capacitated sperm with progesterone; and (iv) progesterone-induced Ca(2+) influx using fura-2AM. RESULTS: Intact sperm and sperm extracts possessed proteasome activity, which was susceptible to inhibition by clasto-lactacystin beta-lactone. Sperm-zona binding was not inhibited by clasto-lactacystin beta-lactone. However, both zona pellucida- and progesterone-induced acrosome reactions were inhibited by clasto lactacystin beta-lactone. The proteasome inhibitor also blocked the sustained phase of the Ca(2+) influx provoked by progesterone but not the peak. CONCLUSION: The human sperm proteasome is involved in the exocytosis of the acrosome, perhaps in events upstream of the plateau phase of the Ca(2+) influx. PMID- 12721179 TI - Inter-individual variability in the morphological assessment of human sperm: effect of the level of experience and the use of standard methods. AB - BACKGROUND: The assessment of human sperm morphology is not devoid of methodological problems, making comparison among laboratories difficult. The main goal of this study was to assess how the experience and compliance to the recommended procedures can modulate it. METHODS: Sixty-two technicians and biologists of varying degrees of andrological experience classified the same 100 sperm in a blind study from images captured on a video microscope and projected onto a screen. David's morphology classification which distinguishes normal and abnormal sperm and calculates a Multiple Anomalies Index (MAI) was used. These sperm were also classified by two trained and experienced biologists using videotaped images to provide reference values. RESULTS: The inter-participant coefficient of variation was 40% for normal sperm, lower for five defects and, 12% for MAI. Experienced participants had the closest results to the reference values. Moreover, participants not routinely using the recommended methodologies, regardless of their experience level, consistently showed marked differences compared with the reference values. CONCLUSIONS: The present data confirm the wide variability in the assessment of normal sperm and show a lower variability for several anomalies and MAI. They underscore the role that experience plays and the importance of following the recommended methodologies for relevant and comparable results. PMID- 12721180 TI - Sperm DNA fragmentation decreases the pregnancy rate in an assisted reproductive technique. AB - BACKGROUND: Standard sperm characteristics are poor predictors of the outcome of IVF treatments. On the contrary, sperm genome quality has been emphasized for several years as playing a major role in early embryogenesis, thus in the success of IVF attempt. METHODS: Sperm DNA fragmentation from a selected group of 104 couples undergoing assisted reproductive techniques (ART) (IVF: n = 50; and ICSI: n = 54) was measured by TUNEL assay and correlated with semen and ART outcomes. RESULTS: A negative correlation was found between sperm characteristics and the proportion of sperm showing DNA fragmentation. For fragmentation >10%, a significant decrease of the fertilization rate was observed. No correlation was found between sperm DNA fragmentation and embryo quality. A high proportion of sperm with fragmented DNA was a pejorative factor to obtain pregnancies when ICSI was performed, but there was no relationship when conventional IVF was performed. CONCLUSIONS: The proportion of sperm with DNA fragmentation appears to be potentially useful as a predictor of ICSI outcome, whereas embryo quality based on morphological criteria, appeared unaffected by DNA fragmentation. PMID- 12721181 TI - Human sperm volume regulation. Response to physiological changes in osmolality, channel blockers and potential sperm osmolytes. AB - BACKGROUND: Volume regulation is an important sperm function because defective sperm cannot negotiate the female tract in an infertile mouse model and swollen human sperm cannot penetrate and migrate through mucus. METHODS AND RESULTS: The size of sperm from 52 donor ejaculates incubated in medium of female tract fluid osmolality (BWW290) was measured by flow cytometry to be identical to that in homologous semen osmolality (289-351 mosmol/kg), indicating effective volume regulation. Inhibition of anticipated regulatory volume decrease in BWW290 by the channel blocker quinine induced size increases and associated kinematic changes measured by computer-aided sperm analysis. Incubation in L-carnitine, myo inositol and taurine did not change sperm volume or kinematics, but the presence of glutamate and K(+) decreased the efficiency of forward progression indicative of volume increase, suggesting them as potential osmolytes for human sperm. Linear regression suggested correlations of changes in cell volume and in kinematic parameters, and the association of faster forward progressive sperm with smaller cell size. CONCLUSIONS: Sperm volume and its regulation may be crucial to natural fertility. The identification of sperm osmolytes, ion channels and mechanisms involved would contribute to the understanding of male infertility and offer a lead for male contraception. PMID- 12721182 TI - Accuracy of sperm-cervical mucus penetration tests in evaluating sperm motility in semen: a systematic quantitative review. AB - BACKGROUND: Our objective was to determine the accuracy of in-vitro sperm penetration into cervical mucus or substitutes in evaluating sperm motility in semen. METHODS: This was a systematic quantitative review of test accuracy studies. The Cochrane library (2000:4), Medline (1966-2001), Embase (1988-2001) and SciSearch (1981-2001) were searched, in addition to manual searches of conference papers and bibliographies of known primary and review articles. Primary studies measuring in-vitro sperm penetration into cervical mucus, or substitutes (i.e. sperm-mucus penetration test, SMPT) and comparing results with sperm motility in semen were included. RESULTS: There were 18 primary diagnostic studies published in 17 papers, involving a total of 2580 samples. Fourteen primary diagnostic tests used vanguard distance as diagnostic criteria (SMPT(vd)) and the pooled likelihood ratio (LR) for positive (LR+) and negative (LR-) tests were 2.29 (1.82-2.87) and 0.52 (0.44-0.63) respectively. Four studies used diagnostic criteria based directly or indirectly on swim-up sperm count per high power field (SMPT(sc)) instead. Their pooled LR+ and LR- were 5.24 (3.36-8.18) and 0.15 (0.06-0.39) respectively. CONCLUSIONS: SMPT(vd) has a low accuracy in the evaluation of sperm motility in semen. However, SMPT(sc) was found to be more accurate. This method of using sperm concentration, instead of vanguard distance, as diagnostic criteria of in-vitro SMPT has potential as a useful laboratory based sperm function test. PMID- 12721183 TI - Characterization of plasma inhibin forms in fertile and infertile men. AB - BACKGROUND: The reciprocal relationship between plasma FSH and inhibin B generally reflects the state of spermatogenesis but data in some settings indicate further complexity in their relationship. Inhibin circulates as a range of higher molecular weight (mol wt) forms of varying bioactivity such that the serum profile of inhibin forms may differ between normal men and those with varying types of spermatogenic failure. The aim of this study was to establish if the inhibin B mol wt distribution was altered in men with infertility. METHODS: The mol wt profiles of inhibin B and free alpha-subunit were determined in plasma of fertile (n = 11) and infertile (n = 17) men using a combined immunoaffinity chromatography, preparative SDS-PAGE and electro-elution procedure and fractions assayed using ELISAs for inhibin B, total inhibin (all forms containing the alpha subunit) and free alpha-subunit. RESULTS: Inhibin B was identified as precursor (60-65 k) and mature (26-30 k) forms in plasma in similar proportions (29%) in fertile men and oligospermic men (25%), but was undetectable in azoospermic men. The free alpha-subunit detected by the pro-alphaC ELISA was identified as both the precursor and processed (pro-alphaC) forms with similar proportions in fertile (8%) and all infertile (4-14%) men. The pro-alphaC ELISA did not detect the precursor forms of inhibin B in plasma while the inhibin B ELISA detected all total inhibin forms following removal of pro-containing forms by immunoabsorption. CONCLUSIONS: (i) the proportions of precursor inhibin B and alpha-subunit forms in the circulation are unchanged in men with spermatogenic disorders indicating there is no alteration of the Sertoli cell inhibin secretory pattern; (ii) these fractionation studies indicate that pro-alphaC and inhibin B ELISAs specifically detect the free alpha-subunit and inhibin B forms present in male plasma. PMID- 12721184 TI - Traffic pollutants affect fertility in men. AB - BACKGROUND: Given the lack of consensus about the effect of traffic-derived pollutants on male fertility, we evaluated semen quality in men occupationally exposed to traffic. METHODS: Semen quality was investigated in 85 men employed at motorway tollgates and in 85 age-matched men living in the same area. Semen, circulating sex hormones, methaemoglobin, sulphaemoglobin, carboxyhaemoglobin, lead (Pb) and zinc (Zn) protoporphyrin were assayed. Environmental carbonium oxide, nitrogen oxide, sulphur oxide and Pb were also measured. RESULTS: Sperm count, and serum levels of FSH, LH and testosterone were within normal range in both groups. Total motility, forward progression, functional tests and sperm kinetics were significantly lower in tollgate workers versus controls. In a subset of tollgate workers with motility below normal, methaemoglobin was inversely correlated with total motility, viability, the hypo-osmotic swelling test, the acridine orange test, the cervical mucus penetration test, linearity, and amplitude of lateral movement of the sperm head, whereas blood levels of Pb were inversely correlated with viability and sperm count. CONCLUSIONS: The finding that blood methaemoglobin and Pb were inversely correlated with sperm parameters indicates that nitrogen oxide and Pb adversely affect semen quality. PMID- 12721185 TI - Multinucleation in cleavage stage embryos. AB - BACKGROUND: The aim was to analyse multinucleation in relation to its incidence in time and in the population, and its correlation with clinical variables, with other morphological characteristics and with the implantation rate of cleavage stage embryos. METHODS: Retrospective analysis of 10 388 cleaved embryos from 1395 consecutive IVF/ICSI cycles in 700 patients between January 1, 1999 and June 30, 2002. RESULTS: Multinucleation was observed in 3491 (33.6%) embryos in 1107 cycles (79.4%) of 609 (87%) patients, more frequently on day 2 than on day 3: 2848 (27.4%) versus 1567 (15.1%) [relative risk (RR) = 1.82; 95% confidence interval (CI) = 1.72-1.92]. Its incidence increased with fragmentation: 31.0, 34.4 and 36.5% for fragmentation /=20 IU/l) male serum FSH and embryo development, manifested as embryos not reaching the morula stage on day 5 (r = 0.387; P < 0.05). After prolonged culture, 23% of embryos reached the blastocyst stage. The pregnancy rates per transfer, and the abortion rates were approximately the same in the day 2 group and the day 5 group (20 versus 20% and 19 versus 18% respectively). After blastocyst transfer, a high clinical pregnancy rate (55%) and a low abortion rate (6%) were achieved, whereas the transfer of arrested embryos provided a low pregnancy rate (2%) and a high abortion rate (100%). If only blastocysts had been transferred on day 5, the clinical pregnancy rate per started cycle would have been approximately the same in both groups (13 versus 16%). CONCLUSIONS: Blastocyst formation is a good indicator of clinical results after ICSI with testicular sperm. PMID- 12721187 TI - Pregnancy following treatment of symptomatic myomas with laparoscopic bipolar coagulation of uterine vessels. AB - BACKGROUND: Laparoscopic bipolar coagulation of uterine vessels (LBCUV) has been employed for women with symptomatic uterine myomas, but its effect on subsequent pregnancy has not been characterized. METHODS: Four-hundred and twenty-three women entered the study between March 1999 and December 2001. Of these, 142 women (33.6%) were under the age of 40 years at the time of LBCUV, 36 of whom (36/142, 25.3%) were sexually active without contraception. In a prospective study of 142 patients (<40 years old) undergoing LBCUV for symptomatic myomas, 15 women became pregnant (17 total pregnancies) and were evaluated by physical and ultrasound examinations. RESULTS: The volume of the dominant myoma was 117.4 +/- 118.4 and 36.8 +/- 56.8 cm(3) before and after LBCUV respectively. Volume of the dominant myoma after pregnancy was 46.2 +/- 76.7 cm(3) (mean +/- SD). There was a significant difference in myoma volume before and after LBCUV (P = 0.002), but no significant difference in myoma volume when comparing post-partum size with post LBCUV size (P = 0.269). Pregnancy outcomes included seven miscarriages in the first trimester and one premature rupture of membrane (PPROM). Although the other pregnancies were regarded as uncomplicated, only two women were delivered of normal neonates as the other seven pregnancies were terminated secondary to patient request. CONCLUSIONS: The pregnancy and term pregnancy rates in sexually active women without contraception were 41.6% (15/36) and 5.6% (2/36) respectively. Because a relatively high rate (7/17, 41.2%) of early miscarriages was observed, we recommend that this procedure be employed only for women who do not desire additional children. PMID- 12721188 TI - Uterine endometrial thermal balloon therapy for the treatment of menorrhagia: long-term multicentre follow-up study. AB - BACKGROUND: Initial reports from observational and randomized trials of uterine endometrial thermal balloon therapy (UBT) suggested good results as judged by return to eumenorrhoea or less and patient satisfaction. Long-term follow-up data remained limited by the small numbers of patients and duration of follow-up. We present long-term (4-6 years) follow-up data from a cohort of women previously treated with UBT for menorrhagia. METHODS: Of the 260 questionnaires sent to women eligible for long-term follow-up from 10 centres, 188 (72%) replies were received. The primary outcome measure was avoidance of hysterectomy. RESULTS: In women who responded to the questionnaire, 25 had undergone hysterectomy and 21 had had repeat ablation. At 4-6 years after UBT, the probability of avoiding hysterectomy was 86% of all women, and of avoiding re-ablation was 88% of non hysterectomized women. Overall, the probability of avoiding any surgery was 75%. Women with an axial or retroverted uterus were at greater risk of hysterectomy or re-ablation. Among the participants, 47% of the non-hysterectomized women were amenorrhoeic, 30% were hypomenorrhoeic, 13.6% were eumenorrhoeic and 8.5% had heavy periods. CONCLUSIONS: This is the first long-term follow-up report of a second-generation endometrial ablation procedure and confirms our initial experience. The high rate of hysterectomy avoidance over 5 years or more is very encouraging for this technology. PMID- 12721189 TI - Vascular system of intramural leiomyomata revealed by corrosion casting and scanning electron microscopy. AB - BACKGROUND: The vascular system of leiomyomata, the most common benign tumours in women, is an important factor controlling development and growth of the tumour. It has not been, however, investigated morphologically using the best currently available technique, corrosion casting combined with scanning electron microscopy. METHODS: Myomatous uteri collected upon autopsy were perfused via afferent vessels with fixative followed by Mercox resin and corroded after polymerization of the resin. The obtained vascular casts visualizing all vessels including capillaries were examined using scanning electron microscopy. RESULTS: The smallest (1-3 mm) fibroids were avascular, in larger ones (<1 cm) a few small vessels invaded the lesion from the periphery. The largest tumours (>1 cm) contained irregular networks of blood vessels with density similar to or lower than that of normal myometrium. Such tumours were surrounded by an extremely dense vascular layer ('vascular capsule') which was the source of larger vessels supplying and draining the tumour. CONCLUSIONS: During development of leiomyoma, the pre-existing blood vessels undergo regression and new vessels invade the tumour from the periphery, where intense angiogenesis, probably promoted by growth factors secreted by the tumour, leads to the formation of a 'vascular capsule' responsible for supply of blood to the growing tumour. PMID- 12721190 TI - The pathogenic role of anti-thyroglobulin antibody on pregnancy: evidence from an active immunization model in mice. AB - BACKGROUND: The presence of antibodies to thyroglobulin (Tg) is associated with fetal loss even in the absence of thyroid dysfunction. The aim of this study was to examine whether active immunization with Tg could elicit anti-Tg autoantibodies and reproductive failure without interfering with thyroid function. METHODS: BALB/c mice that were immunized with human Tg in complete Freund's adjuvant (CFA) or injected with only CFA were studied for the development of antibodies to Tg, T4, dsDNA, ssDNA and cardiolipin. Total T4, free T4 and thyroid-stimulating hormone (TSH) levels were also assessed before and during pregnancy. Percentages of resorbed fetuses (the equivalent to human missed abortion) were compared and autoantibody presence on the placentae and fetuses was examined. RESULTS: Following immunization, high levels of anti-Tg were observed in mice immunized with Tg, compared with mice injected with CFA [0.83 +/ 0.23 versus 0.012 +/- 0.016 respectively; mean +/- SD optical density (OD) at 405 nm; P < 0.001]. The specificity of binding to Tg was confirmed by competition assay. Although total T4 levels were increased in comparison with control mice, this was associated with the presence of antibodies to T4. Indeed, free T4 levels and TSH were similar to control mice. Mice were killed after 14 days of pregnancy. The thyroid function and the histology of the thyroid glands were normal. Increased fetal wastage was found among the Tg-immunized mice compared with the CFA-injected mice (P = 0.04), with lower fetal and placental weights (fetal weights: 194 +/- 4 mg versus 240 +/- 6 mg; placental weights: 105 +/- 2 mg versus 130 +/- 3; P < 0.001 for both). Antibodies to Tg were demonstrated only on the placentae of Tg-immunized mice. CONCLUSION: Immunization with Tg results in the production of Tg antibodies and fetal resorption. These effects occur in the absence of thyroid dysfunction. PMID- 12721191 TI - Changes in endometrial blood vessels in the endometrium of women with hormone replacement therapy-related irregular bleeding. AB - BACKGROUND: Irregular bleeding affects up to 60% of hormone replacement therapy (HRT) users. The mechanism of this bleeding is not understood. Reduced endometrial microvascular integrity appears to underlie breakthrough bleeding in pre-menopausal women and the aim of this study was to establish whether similar changes are seen in HRT users and hence to elucidate a possible mechanism of irregular bleeding. METHODS: Endometrium from 34 HRT users with amenorrhoea, irregular bleeding or regular bleeding was assessed for endometrial endothelial cell density (anti-CD34), number of blood vessels per mm(2), vascular basal lamina components (laminin, collagen IV and heparan sulphate proteoglycan) and in 32 subjects and 23 controls for perivascular smooth muscle alpha (SMA). Findings were compared with a control population of 29 post-menopausal women not using HRT, other sex steroids or tamoxifen and with no vaginal bleeding. Staining intensity was assessed in a blinded fashion in all immunohistochemical studies. RESULTS: Four significant differences in endometrial blood vessels were observed between HRT users and controls: (i) a significantly lower density of endometrial endothelial cells (EC staining for CD34) per mm(2) was present in HRT users compared with controls (P < 0.001); (ii) endothelial cells (EC) were predominantly organized within blood vessels (83%) in controls but in HRT users EC were dispersed in the tissues with only 29% in organized vessels (P <0.001); (iii) supportive perivascular cell SMA was significantly reduced in 23 post menopausal HRT users compared with 23 post-menopausal controls (n = 29, P = 0.013) and (iv) an atrophic or inactive histological pattern of endometrium was more frequently seen in the controls (P < 0.001). CONCLUSIONS: These findings support the hypothesis that exposure to HRT profoundly alters endometrial blood vessels, reducing structural integrity thereby predisposing to irregular bleeding in HRT users. PMID- 12721192 TI - Public opinion regarding oocyte donation in Sweden. AB - BACKGROUND: Oocyte donation has been permitted by Swedish legislation since January 2003. According to the law, offspring have the right to receive identifying information about the donor when they reach a mature age. The aim of the present study was to investigate public opinion regarding different aspects of oocyte donation. METHODS: A study-specific questionnaire regarding attitudes towards aspects of oocyte donation was sent to a randomized sample of 1000 women (73% response) and 1000 men (56% response). RESULTS: A majority of respondents supported treatment with oocyte donation. Seventeen per cent of the women considered donating in the future, whereas 56% of the men would support their partner. While nearly half of the respondents considered that offspring should receive identifying information of the donor, one-third were opposed to this. Overall, women were more positive towards disclosure to the offspring than were men (P < 0.001). CONCLUSIONS: The present results indicate strong support for the use of oocyte donation among a subset of the Swedish population. There was considerable interest among women in donating oocytes anonymously. While a majority advocated openness regarding the donation between parents and child, there was less support for the offspring to have a legal right to receive identifying information about their donor. PMID- 12721193 TI - Choosing identity-release sperm donors: the parents' perspective 13-18 years later. AB - BACKGROUND: A growing number of donor insemination (DI) programmes offer 'open identity' sperm donors, who are willing to have their identity released to adult offspring. We report findings from parents who chose such donors and whose children are now adolescents. METHODS: Using mail-back questionnaires, parents from 45 households (40% headed by lesbian couples, 38% by single women, 22% by heterosexual couples) reported their experience with using an open-identity donor and disclosure about it, as well as their child's plans for donor identity release(sm). RESULTS: Almost no parents regretted using an open-identity donor. Almost all parents had told their child about his or her DI conception early on and reported a neutral to moderately positive impact. Finally, of those who had told, almost all expected their child to obtain the donor's identity. We also discuss differences found between birth mothers and co-parents and among single women, lesbian couples, and heterosexual couples. CONCLUSIONS: Families were relatively open and positive about their use of DI and that their child could obtain the donor's identity. Disclosure did not appear to have a negative impact on the families, regardless of parental sexual orientation and relationship status. PMID- 12721194 TI - Prions, urinary gonadotrophins and recombinant gonadotrophins. PMID- 12721195 TI - Evidence that endometriosis results from the dislocation of basal endometrium? PMID- 12721197 TI - Antiphospholipid antibodies (APA) and recurrent pregnancy loss: treating a unique APA positive population. PMID- 12721199 TI - Adrenal antibiotics detect asymptomatic auto-immune adrenal insufficiency in young women with premature ovarian failure. PMID- 12721201 TI - Sex ratios of births conceived during wartime. PMID- 12721203 TI - Testicular pain following vasectomy: a review of postvasectomy pain syndrome. PMID- 12721204 TI - Managing the risks of prostate disease during testosterone replacement therapy in older men: recommendations for a standardized monitoring plan. PMID- 12721205 TI - Roeing against the wind and wadeing against the current: embryonic stem cell research and the abortion debate. PMID- 12721206 TI - Using the male gamete for assisted reproduction: past, present, and future. PMID- 12721207 TI - Responses to semen analysis CART report. PMID- 12721209 TI - New semen quality scores developed by principal component analysis of semen characteristics. AB - The purpose of this study was to determine whether semen characteristics can be reduced to 2 semen quality (SQ) scores and whether these new scores can help the clinician in assessing the reproductive outcome. A cross-sectional sample of 250 patients seeking infertility treatment were analyzed for semen characteristics. In addition, 177 male-factor patients (prostatitis with infection, n = 40; varicocele, n = 77; varicocele with infections, n = 11; and vasectomy reversal, n = 43) were also assessed. Sperm motion kinetics were measured by computer assisted semen analysis (CASA) (concentration, percent motility, curvilinear velocity [VCL], straight-line velocity [VSL], average path velocity [VAP], linearity [LIN], and amplitude of lateral head displacement [ALH]). Sperm morphology was assessed by both World Health Organization (WHO) guidelines and Tygerberg strict criteria. The principal component analysis model was used to construct an SQ score and a relative semen quality (RQ) score. A separate set of 25 normal donors was included as controls to determine normal ranges of the semen scores. Among the patient samples, SQ and RQ scores (median and 25% and 75% interquartile values) were 89.9, 25.1, and 130.4 and 106.1, 45.2, and 165.9, respectively. The SQ score for the varicocele and varicocele with infection groups was comparable (78.6 +/- 17.4 and 84.8 +/- 20.6) but significantly different from the control (100 +/- 10, P <.001 and.03). Vasectomy reversal patients had an SQ score of 78.2 plus or minus 16.8 that was significantly lower than controls (P <.001). The correlation among semen characteristics allows for the efficient combining of semen measures. The composite scores can summarize overall SQ and quantity. Both SQ and RQ scores provide meaningful information on the quality of semen specimens for the clinician. PMID- 12721208 TI - Evidence for a role of glycogen synthase kinase-3 beta in rodent spermatogenesis. AB - Glycogen synthase kinase-3 beta (GSK-3 beta) regulates cell metabolism, cell cycle, and cell fate through the phosphorylation of a diverse array of substrates. Herein, we provide evidence that supports a role for GSK-3 in mammalian meiosis and spermatogenesis. Immunostaining of testis sections showed that while GSK-3 alpha was ubiquitous in the seminiferous tubules, GSK-3 beta was expressed in premeiotic type B spermatogonia, in both meiotic preleptotene and leptotene spermatocytes, as well as in Sertoli cells in both the mouse and rat. Thus, GSK-3 beta is expressed in germ cells entering meiosis. In addition, intense immunoreactivity was detected in rat step 6 though 11 spermatids. In situ hybridization (ISH) in rat testis confirmed the immunostaining pattern in leptotene and spermatids and showed a GSK-3 beta messenger RNA (mRNA) signal in some pachytene spermatocytes. The restricted pattern of expression suggests cell specific regulation of Gsk-3 beta mRNA. To determine whether GSK-3 is required for meiosis entry, rat stage VIIa seminiferous tubule segments were cultured with selective small-molecule GSK-3 inhibitors. These compounds markedly and dose dependently suppressed meiotic synthesis (S)-phase DNA. Since a yeast GSK-3 homolog, Rim11p (regulator of inducer of meiosis), is pivotal to meiosis entry, we tested whether GSK-3 beta complements Rim11p function in meiosis. Rim11p phosphorylates transcription factors Ume6p (unscheduled meiotic gene expression) and Ime1p (inducer of meiosis) to induce meiosis entry. Overexpression of murine GSK-3 beta in a rim11 mutant yeast failed to rescue the sporulation defect. Our finding that GSK-3 beta interacted only with Ume6p but not with IME1 in a yeast 2 hybrid assay suggests that noncomplementation reflects partial divergence in substrate specificity. This work provides the basis for future studies of GSK-3 beta signaling in mammalian meiosis and spermatogenesis. PMID- 12721210 TI - The use of novel semen quality scores to predict pregnancy in couples with male factor infertility undergoing intrauterine insemination. AB - The purpose of this study was to determine whether 2 new semen quality (SQ) scores could predict pregnancy in patients undergoing intrauterine insemination (IUI) for male-factor infertility and whether an overall score could help in counseling these couples with assisted reproductive technique (ART) options. Ninety-three couples with male-factor infertility were examined for semen analysis. Samples were prepared by density gradient separation (47% and 90%), and IUI was performed. On the basis of the 2 semen scores (SQ and relative quality [RQ]), the IUI-semen pregnancy score (IUI-SPS) was calculated. Of the 192 IUI cycles, 14% (27 of 192) resulted in pregnancy. Both prewash SQ and RQ scores were significantly related to pregnancy (P =.02 and P <.001), as was the postwash RQ score (P <.001). Of the IUI cycles in which the postwash RQ score was greater than 125, 40% (13 of 32) resulted in pregnancy compared to 9% of cycles (14 of 160) in which the postwash SQ score was less than 125. The prewash IUI-SPS score was significantly related to IUI-induced pregnancy (P <.001). Both the pre- and postwash SQ and RQ scores can predict pregnancy in male infertility patients undergoing IUI. Patients with an IUI-SPS less than 150 may be advised to seek in vitro fertilization (IVF), whereas those with an IUI-SPS greater than 150 may be advised to seek IUI. PMID- 12721211 TI - Rapid disappearance of spermatozoa after vasal occlusion in the dog. AB - The reproductive system of the male dog is unusual in that the seminal vesicles and bulbourethral glands are absent. Therefore, we chose the dog as a model to evaluate the effect of seminal vesicles on clearance of spermatozoa from the male reproductive tract after vasal occlusion. Thirty adult male beagle dogs with ejaculates containing at least 500 x 10(6) sperm with greater than 90% motility were used for this study. The dogs' vasa were occluded percutaneously using a Vasocclude clip-applying device through a small scrotal puncture site. Dogs were ejaculated and semen analysis was performed before and after vas occlusion. The first 24 dogs were completely azoospermic 1 week following vas occlusion. In order to explain these unanticipated results, an additional 6 dogs were evaluated to determine the specific time course of sperm disappearance from the ejaculate at 1, 3, 5, and 7 days after vas occlusion. The results revealed that spermatozoa were almost completely absent within 1 day after vas occlusion (99.9% reduction, X = 1.0 + 1.1 x 10(6) sperm per milliliter at 1 day, P <.0005 vs prevas occlusion). The rapid elimination of spermatozoa after vas occlusion indicates that sperm transit rapidly through the vas deferens in dogs. Therefore, the delayed clearance of spermatozoa from the ejaculate in humans may be due to sperm storage in the seminal vesicles. PMID- 12721212 TI - Effect of azorellanone, a diterpene from Azorella yareta Hauman, on human sperm physiology. AB - Previous studies have shown that cyclic terpenes extracted from plants decrease sperm motility and concentration in rats. In this work, we studied the effect 13 alpha-hydroxy-7-oxoazorellano (azorellanone), a cyclic diterpene extracted from Azorella yareta Hauman, on in vitro human sperm physiology. Sperm aliquots, capacitated for 4.5 or 20 hours, were incubated for 15 minutes with different concentrations of azorellanone. Then, the effects of azorellanone on sperm motility, viability, binding to the human zona pellucida, progesterone-induced acrosome reactions and increase in intracellular Ca(2)(+) concentration, and trypsin and chymotrypsin-like protease activities were evaluated. Sperm motility was evaluated according to World Health Organization (WHO) guidelines; sperm viability with the supravital dye Hoescht 33258; sperm-zona binding with the hemizona assay; progesterone-induced acrosome reaction with fluorescent lectin; intracellular Ca(2)(+) level with fura 2; and protease activity with the synthetic substrates N-t-Boc-Gln-Ala-Arg-Amido-4-methylcoumaryn and Succinyl-Leu Leu-Val-Tyr-Amido-4-methylcoumaryn. The results obtained indicate that azorellanone inhibited sperm motility in a concentration-dependent manner at 0.15, 1.5, and 3 mM, while sperm viability was only inhibited at 3 mM. Treatment with azorellanone significantly inhibited sperm-zona binding, progesterone induced acrosome reactions, and intracellular Ca(2)(+) concentration. Treatment of free-swimming sperm with azorellanone did not affect protease activity; however, the incubation of sperm extracts with azorellanone significantly inhibited both trypsin-like and chymotrypsin-like protease activities. In conclusion, azorellanone has a significant effect on the different parameters that characterize human sperm physiology. PMID- 12721213 TI - Spinal cord contusion impairs sperm motility in the rat without disrupting spermatogenesis. AB - Our previous studies demonstrated various abnormalities in spermatogenesis after spinal cord injury (SCI) in cord-transected rats. In this study, we examined whether abnormalities in spermatogenesis in SCI rats were related to the degree of SCI. We used spinal cord-contused (SCC) rats as a model. Adult male Sprague Dawley rats were subjected to various degrees of cord contusion caused by the weight of a rod dropped from different heights (12.5, 25, 50, and 75 mm) using a New York University IMPACTOR. Testicular histology revealed persistent complete spermatogenesis in all SCC rats 4, 8, or 14 weeks after cord contusion regardless of the extent of SCI. Northern blot complementary DNA (cDNA) hybridization revealed transient but significant decreases in the levels of Sertoli cell specific transcripts in SCC rats. In addition, levels of messenger RNA (mRNA) transcripts for germ cell-specific transition protein-2 and protamine-1 were consistently decreased in these rats. Such effects were related to the height of the weight drop and were associated with reduced levels of mRNA for cyclic adenosine monophosphate (cAMP) responsive element modulator (CREM). These results demonstrated specific effects of SCI on spermiogenesis and were consistent with altered cAMP signaling in testicular cells after SCI. Sperm motility was also significantly decreased in SCC rats and was related to the height of weight drop. Normal sperm motility recovered only in those rats injured by weight drop from 12.5- and 25-mm heights. In summary, current results demonstrate persistent abnormalities in spermiogenesis and sperm motility in rats that suffered spinal cord contusion by weight drop. Such effects were related to the height of the weight drop and thus to the extent of SCI. PMID- 12721214 TI - Effects of medical or surgical castration on erectile function in an animal model. AB - The goal of this study was to investigate the effects of medical castration (luteinizing hormone-receptor hormone [LH-RH] agonist treatment) or surgical castration on erectile function in an animal model. New Zealand White male rabbits were either kept intact (control); surgically orchiectomized; or treated for 2, 4, or 8 weeks with the LH-RH agonist leuprolide acetate (107 microg/kg/mo). At 2 weeks, plasma testosterone levels of orchiectomized and leuprolide acetate-treated animals were 12.8% and 57.4% of intact control animals, respectively. Erectile function was assessed by continuously recording systemic arterial pressure (SAP) and intracavernosal blood pressure (ICP) and determining the ICP:SAP ratios in response to electrical stimulation of the pelvic nerve at varying frequencies (2.5-32 Hz). Androgen deprivation by surgical (orchiectomy) or medical (leuprolide acetate) castration reduced ICP at all frequencies tested but did not alter SAP. Administration of the phosphodiesterase type 5 inhibitor vardenafil (10 microg/kg) did not enhance ICP in surgically orchiectomized or leuprolide acetate-treated animals. Nitric oxide synthase and arginase activities in the corpus cavernosum were not significantly altered by surgical or medical castration. Further, Masson trichrome staining of erectile tissue from androgen-ablated animals showed a reduction in smooth muscle content. These data demonstrate that androgen deprivation achieved by surgical or medical castration adversely affects penile hemodynamics and erectile function without producing significant changes in the activities of nitric oxide synthase or arginase. We conclude that androgen deprivation produces structural alterations in the corpus cavernosum leading to corporal veno-occlusive dysfunction. PMID- 12721216 TI - Relationship between the characteristics of epididymal red deer spermatozoa and penetrability into zona-free hamster ova. AB - A heterologous (zona-free hamster oocytes) in vitro fertilization (IVF) system was used to evaluate the relationship between sperm factors and penetration capacity of epididymal red deer spermatozoa. The sperm parameters evaluated in 36 sperm samples obtained postmortem from stags selectively shot during the rutting season were sperm motility, functional integrity of plasma membrane by means of the hypo-osmotic swelling test (HOST), and, simultaneously, viability and acrosomal status via a triple-stain technique. Zona-free hamster oocytes were used to evaluate the capacity of the different sperm assays to predict in vitro penetration. In order to increase the variability in sperm quality, we recovered samples from stags at different intervals between the death of the male and the collection of the genitalia. All measures of sperm quality declined progressively (P <.001) with increasing intervals between death and sample collection. In addition, many sperm parameters were related to penetration ability in vitro. Subsequently, sperm samples were rearranged in 2 categories according to the interval that had elapsed between death and the collection of the genitalia (group 1, short interval = 0-12 h; group 2, large interval = 18-40 hours). When samples were grouped, less correlation achieved significance, especially for group 1, than when samples were not divided. Also, correlation between the number of sperm per oocyte and sperm parameters for group 1, which had the highest values of sperm quality, failed to reach significance. It is concluded that the classical parameters accepted in assessing the viability of deer spermatozoa can be good predictors of the penetrating ability of the spermatozoa when satisfactory in vitro conditions are used for the development of the IVF system. Also, this study demonstrates that compatible heterologous gamete interaction allows thorough assessment of sperm function in a wild deer. PMID- 12721215 TI - Sperm mitochondrial mutations as a cause of low sperm motility. AB - We report the unique case of a 28-year-old man who, in spite of having a varicocele and a sperm concentration of 5 million/mL, of which 10% were motile and 20% had normal forms (oligoasthenoteratozoospermia [OAT]), was fertile. This was confirmed by paternity testing using 16 autosomal and 6 Y-chromosomal short tandem repeat (STR) loci. An analysis of mitochondrial genes that included cytochrome oxidase I (COI), cytochrome oxidase II (COII), adenosine triphosphate synthase6 (ATPase6), ATPase8, transfer ribonucleic acid (tRNA) serine I, tRNA lysine, and NADH dehydrogenase3 (ND3) revealed, for the first time, 9 missense and 27 silent mutations in the sperm's mitochondrial DNA (mtDNA) but not in the DNA from the blood cells. There was a 2-nucleotide deletion in the mitochondrial COII genes, introducing a stop codon, which might be responsible for low sperm motility. PMID- 12721217 TI - Hormonal regulation of bovine secretory proteins derived from caput and cauda epididymal epithelial cell cultures. AB - The goal of this study was to investigate the effect of hormones (testosterone, dihydrotestosterone [DHT], and hydrocortisone) on the protein secretion of caput and cauda epididymal epithelial cells cultured in principal cell medium (PCM). A confluent monolayer of caput and cauda epididymal epithelial cells was obtained from serum-containing PCM in the presence or absence of hormones after 7 days of culture at 38.5 degrees C (5% CO(2) in air). The protein secretion of epididymal epithelial monolayers incubated in serum-free PCM for 3 days was examined. The secreted proteins were separated by 2-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (2D SDS-PAGE). A comparison of the different protein patterns showed 61 spots, of which 11 were secreted only in the presence of hormones, 3 appeared to show hormone-related changes, and 25 were region specific. Most of these secreted proteins were low-molecular-weight acidic proteins. To obtain evidence of the epididymal origin of the secreted proteins, proteins present in caput and cauda epididymal plasma were analyzed. In conclusion, our data indicate that hormones influence the synthesis of a number of caput and cauda epididymal proteins. Some of these proteins could be important for improving our understanding of spermatozoa maturation and storage and their acquisition of fertilizing ability. PMID- 12721218 TI - Regulation of sulfated glycoprotein-1 and cathepsin D expression in adult rat epididymis. AB - Endocytosis, whereby proteins are internalized from the epididymal lumen to be eventually degraded in lysosomes, is one of the major functions of the epididymal epithelial cells in maintaining a proper luminal milieu conducive for sperm maturation. In the present study, using light microscope immunocytochemical methods, we examined the regulation of 2 lysosomal enzymes, sulfated glycoprotein 1 (SGP-1) and cathepsin D, in adult rat epididymides fixed in Bouin fixative and embedded in paraffin. After orchidectomy (O) with or without testosterone (T) supplementation, efferent duct ligation (EDL), or hypophysectomy (H), lysosomes of principal cells were intensely reactive with the anti-SGP-1 antibody, as were narrow, clear, and basal cells, with staining patterns similar to that of control animals. These experimental procedures also had no effect on cathepsin D expression in all cell types, except for clear cells of the corpus and cauda epididymidis, which after orchiedectomy and hypophysectomy, became intensely reactive, unlike their completely unreactive state in control animals. In O+T animals, as well as in EDL animals, clear cells remained unreactive. These data taken together suggest that expression of SGP-1 is not under the control of testicular or pituitary factors, as is also the case for cathepsin D expression by principal, narrow, and basal cells. However, specific inhibition of cathepsin D expression by testosterone or one of its metabolites appears to occur in clear cells of the corpus and cauda epididymidis. Furthermore, in addition to small, typical lysosomes, principal cells also revealed large supranuclear and infranuclear spherical structures that were immunoreactive with both anti-SGP-1 and anti-cathepsin D antibodies, suggesting their lysosomal nature. With electron microscopy, these structures appeared electron-lucent and contained membranous profiles embedded in an electron-dense, granular background. Such images suggest that the various experimental procedures adversely affect the expression of several other lysosomal enzymes in principal cells, leading to a lysosomal phenotype similar to that observed in various lysosomal storage diseases. PMID- 12721219 TI - Specific order in the appearance of protein tyrosine phosphorylation patterns is functionally coordinated with dog sperm hyperactivation and capacitation. AB - The aims of the present study were to characterize a slow capacitation system that records initial changes in the sperm membrane state, and, using a canine model, to order the specific protein tyrosine phosphorylation signaling in the sequence of capacitational events and to associate them with hyperactivated motility. Dog sperm washed through Percoll were incubated in complete bicarbonate Tyrode medium for 6 hours in 5% CO(2). Capacitation was evaluated using chlortetracycline staining. Tyrosine phosphorylation patterns were assessed by immunocytochemistry. Parallel to this, a computer-assisted motility analysis was performed. Significant changes in the percentage of capacitated and acrosome reacted cells were first observed after 90 minutes, increasing in a linear manner during further incubation (P <.05). Changes in the percentage of capacitated cells were accompanied by motility changes. During incubation, a strictly sequential phosphorylation of sperm tail (midpiece, principal piece, and end piece) and head proteins was observed. According to an analysis of kinetics, phosphorylation of head proteins occurred after the tail became completely phosphorylated. Changes in head phosphorylation progressed at the same rates as capacitation and acrosome reaction. Sperm motility, curvilinear velocity, average path velocity, straight line velocity, and lateral head displacement were correlated positively or negatively with phosphorylation of midpiece or end piece proteins, respectively. The bicarbonate-stimulated increases in cyclic adenosine monophosphate levels and changes in protein phosphatase activity may be involved in the signaling system that controls membrane changes and motility in dog sperm. Phosphorylation kinetics of sperm proteins are potentially useful for diagnostic purposes to characterize the response of individual males to fertilizing conditions. PMID- 12721221 TI - Male genital tract inflammation: The role of selected interleukins in regulation of pro-oxidant and antioxidant enzymatic substances in seminal plasma. AB - Human semen contains spermatozoa as well as populations of round nonspermatozoal cells primarily consisting of leukocytes. Activation of white blood cells present in the seminal plasma during genital tract inflammation and cellular reactions against microbial agents may provoke a release of a variety of products such as cytokines and reactive oxygen species. The aim of this study was to evaluate whether a panel of selected cytokines (interleukin [IL]-1 beta, IL-6, IL-8, and tumor necrosis factor-alpha [TNF alpha]) detectable in seminal plasma during male genital tract inflammation could be considered as mediators between altered semen parameters and changed levels of pro-oxidant and antioxidant substances. Studies using chemiluminometric, spectrophotometric, and enzyme-linked immunosorbent assay methods indicate that proinflammatory cytokines such as IL-1 beta, IL-6, IL 8, and TNF alpha may modulate pro-oxidant and antioxidant activities in the male genital tract. The data also suggest that the function of pro-oxidant and antioxidant systems in semen may directly influence basic semen parameters. The elevated numbers of leukocytes present in semen during male genital tract inflammation without an associated contribution of cytokines and semen antioxidant capacity appear to be of little prognostic value in evaluating male fertilization potential. PMID- 12721220 TI - Marked differences in protamine content and P1/P2 ratios in sperm cells from percoll fractions between patients and controls. AB - The various sperm cell types present in a simple ejaculate differ in their motility and morphology. However, little is known about the nuclear maturity of these sperm cells and their relationship with morphological and motile characteristics. Protamines are considered a good marker of sperm nuclear maturity since they are added to the DNA in the last stage of spermatogenesis. We have analyzed the P1/P2 ratio and the protamine content of subpopulations of human spermatozoa at different stages of maturation, isolated by density gradient centrifugation of ejaculated spermatozoa obtained from 3 groups of patients from our Assisted Reproduction Unit: 10 men of proven fertility, 12 oligozoospermic men, and 13 asthenozoospermic men. Four different fractions (F2-F5) were collected from the top to the bottom of the Percoll gradient. Differences in the motion and morphology were found between the fractions in each of the groups studied, with fraction F5 being the one with the best morphology and motility. However, no significant differences in the P1/P2 ratio were found between fractions within the same group of samples, indicating that the P1/P2 ratio and the amount of protamines are relatively independent of the morphology and motility of sperm cells. In contrast, statistically significant differences were found in the P1/P2 ratio and in the relative amount of protamines between the 3 groups. PMID- 12721222 TI - DNA surveillance: web-based molecular identification of whales, dolphins, and porpoises. AB - DNA Surveillance is a Web-based application that assists in the identification of the species and population of unknown specimens by aligning user-submitted DNA sequences with a validated and curated data set of reference sequences. Phylogenetic analyses are performed and results are returned in tree and table format summarizing the evolutionary distances between the query and reference sequences. DNA Surveillance is implemented with mitochondrial DNA (mtDNA) control region sequences representing the majority of recognized cetacean species. Extensions of the system to include other gene loci and taxa are planned. The service, including instructions and sample data, is available at http://www.dna surveillance.auckland.ac.nz. PMID- 12721223 TI - Reliable noninvasive genotyping: fantasy or reality? AB - Noninvasive genotyping has not gained wide application, due to the notion that it is unreliable, and also because remedial measures are time consuming and expensive. Of the wide variety of noninvasive DNA sources, dung is the most universal and most widely used in studies. We have developed collection, extraction, and amplification protocols that are inexpensive and provide a high level of success in amplifying both mitochondrial and nuclear DNA from dung. Here we demonstrate the reliability of genotyping from elephant dung using these protocols by comparing results from dung-extracted DNA to results from blood extracted DNA. The level of error from dung extractions was only slightly higher than from blood extractions, and conducting two extractions from each sample and a single amplification from each extraction was sufficient to eliminate error. Di , tri-, and tetranucleotide loci were equally reliable, and low DNA quantity and quality and PCR inhibitors were not a major problem in genotyping from dung. We discuss the possible causes of error in genotyping with particular reference to noninvasive samples and suggest methods of reducing such error. PMID- 12721224 TI - Inbreeding, microsatellite heterozygosity, and morphological traits in Lipizzan horses. AB - While the negative effects of inbreeding and reduced heterozygosity on fecundity and survival are well established, only a few investigations have been carried out concerning their influence on morphological traits. This topic is of particular interest for a small and closed population such as the Lipizzan horse. Thus, 27 morphological traits were measured in 360 Lipizzan mares and were regressed on the individual inbreeding coefficients, as well as on the individual heterozygosity and mean squared distances (mean d(2)) between microsatellite alleles within an individual. Both individual heterozygosity and mean d(2) were based on 17 microsatellite loci dispersed over 14 chromosomes. The results obtained by multivariate analysis reveal significant effects of stud (P <.0001), age at measurement (P <.0001), and mean d(2) (P =.0143). In univariate analyses, significant associations were obtained between length of pastern-hindlimbs and inbreeding coefficient (P <.01), length of cannons-hindlimb and mean d(2) (P <.01), and length of neck and mean d(2) (P <.001). After adjustment of single test P values for multiple tests (Hochberg's step-up Bonferroni method), only the association of the length of neck and mean d(2) remained significant (P =.0213). Thus, no overall large effects of inbreeding, microsatellite heterozygosity, and mean d(2) on morphological traits were observed in the Lipizzan horse. PMID- 12721225 TI - Population Structure of Anopheles gambiae in Africa. AB - The population structure of Anopheles gambiae in Africa was studied using 11 microsatellite loci in 16 samples from 10 countries. All loci are located outside polymorphic inversions. Heterogeneity among loci was detected and two putative outlier loci were removed from analyses aimed at capturing genome-wide patterns. Two main divisions of the gene pool were separated by high differentiation (F(ST) > 0.1). The northwestern (NW) division included populations from Senegal, Ghana, Nigeria, Cameroon, Gabon, Democratic Republic of Congo (DRC), and western Kenya. The southeastern (SE) division included populations from eastern Kenya, Tanzania, Malawi, and Zambia. Inhospitable environments for A. gambiae along the Rift Valley partly separate these divisions. Reduced genetic diversity in the SE division and results of an analysis based on private alleles support the hypothesis that a recent bottleneck, followed by colonization from the NW populations shaped this structure. In the NW division, populations possessing the M rDNA genotype appeared to form a monophyletic clade. Although genetic distance increased with geographic distance, discontinuities were suggested between certain sets of populations. The absence of heterozygotes between sympatric M and S populations in the DRC and the high differentiation in locus 678 (F(ST)>0.28) contrasted with low differentiation in all other loci (-0.02 or = 500 U/L was associated with increased risk of failure in multivariate analysis. CONCLUSION: PMLBL mainly is found in adolescents. Dose-intense chemotherapy including high dose methotrexate yields a pEFS at 5 years of 0.70 (SE, 0.08). LDH is of prognostic value in pediatric patients with PMLBL. PMID- 12721256 TI - Intensification of therapy for children with lower-risk acute lymphoblastic leukemia: long-term follow-up of patients treated on Children's Cancer Group Trial 1881. AB - PURPOSE: From December 1988 through December 1992, the Children's Cancer Group (CCG) conducted a randomized trial (CCG-1881) designed to evaluate the impact of adding a single delayed intensification phase of therapy to standard therapy for patients with newly diagnosed low-risk acute lymphoblastic leukemia (ALL). PATIENTS AND METHODS: Patients (n = 778) with newly diagnosed ALL, 2 to 9 years of age at diagnosis with an initial WBC count less than 10,000/microL, were eligible for this protocol. All patients received induction, consolidation, and interim maintenance phases of therapy over the first 16 weeks. At week 16, patients remaining in remission were randomly assigned to receive or not receive a single 7-week delayed intensification (DI) phase of therapy. Maintenance therapy was given in lieu of or after DI, with total duration of therapy approximately 3 years for boys and 2 years for girls. RESULTS: Patients randomized to receive DI experienced fewer relapse events in all categories. Kaplan-Meier life-table estimates for continuous complete remission (CCR) at 7 years for the randomized regimens were 77% (SE, 2.4%) for the standard regimen and 83% (SE, 2.7%) for the DI regimen (P =.072). The only prognostic factor of significance post-randomization in this selected low-risk population was the day 14 marrow response (P =.0001). CONCLUSION: The addition of a single DI phase of therapy was well tolerated and augmented 7-year CCR by 6% (SE of the difference, 3.3%), resulting in 26% fewer adverse events. Overall survival for eligible patients at 7 years is 90% (SE, 1.2%). PMID- 12721257 TI - CNS-directed therapy for childhood acute lymphoblastic leukemia: Childhood ALL Collaborative Group overview of 43 randomized trials. AB - PURPOSE: A collaborative meta-analysis was performed to clarify the relative effects on relapse and survival of different types of therapies directed at the CNS in childhood acute lymphoblastic leukemia. MATERIALS AND METHODS: Data were sought for each individual patient in all trials started in or before 1993 that included unconfounded randomized comparisons of such treatments. Log-rank survival analyses were performed for each trial, and overall results for groups of trials addressing similar questions were obtained from the totals of the observed minus expected number of events and their variances. RESULTS: Radiotherapy and long-term intrathecal therapy gave similar outcomes, with no significant difference in event-free survival despite random assignment of treatment to 2,848 patients, 1,001 of whom suffered relapse or death. Intravenous methotrexate reduced non-CNS rather than CNS relapses, and hence, the addition of intravenous methotrexate to a treatment regimen including radiotherapy or long term intrathecal therapy improved event-free survival, with a 17% reduction in the event rate (95% confidence interval, 6% to 27%; P =.003). The event-free survival at 10 years in these trials was 61.9% without intravenous methotrexate and 68.1% with intravenous methotrexate. There was no significant difference in survival (14% death rate reduction; P =.09). There were insufficient randomly assigned patients to adequately address other questions, such as effect of different doses. No evidence was found of differences, between trials or between subgroups of different types of patients, in the relative effects of treatment. CONCLUSION: Radiotherapy can be replaced by long-term intrathecal therapy. Intravenous methotrexate gives some additional benefit by reducing non-CNS relapses. PMID- 12721258 TI - Repp86 expression and outcome in patients with neuroblastoma. AB - PURPOSE: Given the well-known challenges of neuroblastoma prognosis, we investigated whether the expression of restrictedly expressed proliferation associated protein of 86 kDa theoretical molecular mass (repp86), a proliferation associated protein expressed in S, G2, and M phases of the cell cycle, correlates with the clinical outcome in patients with neuroblastoma. PATIENTS AND METHODS: 161 children with different stages of neuroblastoma were studied; the median follow-up time was 72.8 months. The patients were staged according to the International Neuroblastoma Staging System, and histologic grading of the tumors was performed according to the criteria of Hughes and those of the International Neuroblastoma Pathology Classification. The MYCN gene copy number was determined by Southern blot analysis or fluorescence in situ-hybridization, and repp86 expression was assessed immunohistochemically by means of monoclonal antibody Ki S2 on paraffin sections from archival tumor samples. RESULTS: A repp86 labeling index (RI) of more than 10% positive tumor cells significantly predicted a shortened disease-free interval and an increased tumor mortality (both P <.0001). Moreover, the RI allowed the identification of patients with favorable and adverse prognosis in subsets defined by stage, grade, age, and MYCN status. In a multivariate analysis, the RI emerged as the most important predictor of event free and disease-specific survival with hazard ratios of 11.7 and 10.5, respectively (both P <.0001). CONCLUSION: It seems that repp86 expression is closely associated with the biologic behavior of neuroblastoma. Assessment of the RI might, therefore, considerably refine prognostic models. PMID- 12721259 TI - Phase III study of cyclophosphamide, doxorubicin, and fluorouracil (CAF) plus leucovorin versus CAF for metastatic breast cancer: Cancer and Leukemia Group B 9140. AB - PURPOSE: To determine whether biochemical modulation with LV (leucovorin) enhances the efficacy of CAF (cyclophosphamide, doxorubicin, and fluorouracil) against metastatic breast cancer. PATIENTS AND METHODS: Women with histologically confirmed stage IV breast cancer, Cancer and Leukemia Group B (CALGB) performance status 0 to 2, and no prior chemotherapy for metastatic disease were randomly assigned to receive CAF (cyclophosphamide 500 mg/m2 day 1, doxorubicin 40 mg/m2 day 1, and fluorouracil [FU] 200 mg/m2 intravenous bolus days 1 to 5) with or without LV (LV 200 mg/m2 over 30 minutes days 1 to 5 given 1 hour before FU). RESULTS: Two hundred forty-two patients were randomly assigned to treatment; 124 patients had visceral crisis and 40 patients had a CALGB performance status score of 2. The median follow-up was 6 years. The two study arms were similar with regard to serious adverse events; four patients died from treatment-related causes, two patients on each study arm. Predictive variables for time to treatment failure and survival were visceral disease and performance status. The overall response rate was 29% for CAF versus 28% for CAF plus LV. The median time to treatment failure (9 months) and median survival (1.7 years) did not differ by treatment arm. CONCLUSION: Modulation of CAF with LV improved neither response rates nor survival among women with metastatic breast cancer, compared with CAF alone. Multivariate analyses confirmed the prognostic importance of performance status and visceral crisis. However, the overall and complete response rates, response durations, time to treatment failure, and survival were the same in the two treatment arms. PMID- 12721260 TI - Natural history of more than 20 years of node-positive primary breast carcinoma treated with cyclophosphamide, methotrexate, and fluorouracil-based adjuvant chemotherapy: a study by the Cancer and Leukemia Group B. AB - PURPOSE: Breast cancer heterogeneity dictates lengthy follow-up to assess outcomes. Efficacy differences for three regimens that are based on adjuvant cyclophosphamide, methotrexate, and fluorouracil (CMF) are presented in this article, but cancer recurrence sites, time of relapse, subsequent primary cancers, and causes of death in the natural history of node-positive breast cancer are emphasized. PATIENTS AND METHODS: Beginning in 1975, 905 patients with node-positive cancer were randomly assigned to receive CMF or two regimens of CMF plus other agents. Median follow-up is 22.6 years. The natural-history analysis was performed on a subset of 814 patients. RESULTS: Eighty percent of the 599 women known to have died, died of metastatic breast cancer. Only 8.5% of the deceased women died of a cause other than breast cancer, a second or third cancer, or adjuvant chemotherapy toxicity. One hundred five women (12.8%) developed other primary cancers, with 49 (46.6%) occurring in the contralateral breast. Therapeutic efficacy differences of the CMF regimens reported earlier have been maintained more than 20 years later. For certain subsets, the five-drug regimen had advantages over CMF. Bone was the most common recurrence site. The longest interval to relapse has been 23.5 years, and 18% of those who relapsed did so more than 10 years later. CONCLUSION: Despite adjuvant chemotherapy, a large majority (80%) of women with node-positive breast cancer die of the disease, and many recurrences develop more than 10 years later. CMF plus vincristine and prednisone provides a benefit compared with CMF, but the magnitude varies with the number of involved nodes. Outcome trends in earlier analyses of this study were maintained even years later. PMID- 12721261 TI - Side effects of adjuvant endocrine treatment in premenopausal breast cancer patients: a prospective randomized study. AB - PURPOSE: To compare the effect of adjuvant endocrine therapies with and without chemotherapy on physical symptoms, anxiety, and depressive symptoms in premenopausal women with breast cancer in a randomized clinical trial (the Zoladex in Premenopausal Patients trial). PATIENTS AND METHODS: The patients were randomly assigned to goserelin, goserelin plus tamoxifen, tamoxifen alone, or no endocrine therapy. The duration of the endocrine treatment was 2 years. The groups were observed for 3 years after primary treatment (ie, during 2 years of active treatment as well as 1 year after cessation of the adjuvant endocrine therapy). All patients with node-positive disease received adjuvant chemotherapy with cyclophosphamide, methotrexate, and fluorouracil (CMF), which was given concurrently with the endocrine treatment. RESULTS: Patients treated with CMF typically reported higher levels of physical symptoms than did patients who did not receive CMF. It was only among patients who did not receive chemotherapy that the endocrine treatment had differential effects. Goserelin was most burdensome and resulted in similar symptom levels as those of CMF, whereas the side effects of tamoxifen alone were milder. After cessation of the endocrine treatment, the side effects diminished in patients who had not received CMF, whereas patients treated with CMF reported ongoing problems at the 3-year follow-up. In contrast, anxiety and depressive symptoms were not significantly affected by endocrine treatment or chemotherapy during the 3 years of assessment. CONCLUSION: Goserelin and tamoxifen resulted in menopausal symptoms, but these symptoms were reversible. However, women treated with CMF experienced physical symptoms throughout the whole study period. PMID- 12721263 TI - Trends in clinical trials reports in common cancers between 1989 and 2000. AB - PURPOSE: Cancer clinical trials can be dichotomized into pilot (phase I and phase II) and randomized controlled clinical trials (RCTs). The best data source for evidence-based medicine is from RCTs. However, many patients prefer to enroll onto pilot trials, and many investigators prefer to conduct or refer their patients to pilot trials. This exploratory study sought to describe the epidemiologic patterns of clinical trial reports in common cancers. METHODS: A structured review was conducted of MEDLINE citations of all English clinical trials reports published between 1989 and 2000 in breast, lung, colorectal, prostate, and female genital cancers, plus leukemias and lymphomas. Each report was classified by design (RCT, pilot, or other) and country. The abstracts of RCTs were reviewed for sample size. Reports addressing screening or prevention were excluded. RESULTS: A total of 12,035 reports, of which 3,560 were from RCTs, were found. The annual growth in RCT reports in breast, colorectal, and prostate cancer was significant (range, 4.8% to 8.5% per year) but was insignificant in leukemias, lymphomas, and female genital and lung cancers (range, 0.1% to 4.3% per year). Within each cancer, the average sample size per report did not change during the 12 years. Nonrandomized trial reports increased on average 15.1% per year (range, 10.1% to 23.2%). The United States accounted for 30% of all RCT reports and 45% of pilot trial reports. CONCLUSION: The faster growth in nonrandomized compared with RCT reports may reflect rapid advances in cancer biology or different structural, commercial, and financial incentives, especially in the United States compared with Europe. Unless additional studies show evidence of an increase in their quality, the modest growth in RCT reports may limit future evidence-based cancer care. PMID- 12721262 TI - Dose escalation of carmustine in surgically implanted polymers in patients with recurrent malignant glioma: a New Approaches to Brain Tumor Therapy CNS Consortium trial. AB - PURPOSE: This New Approaches to Brain Tumor Therapy CNS Consortium study sought to determine the maximum-tolerated dose (MTD) of carmustine (BCNU) that can be implanted in biodegradable polymers following resection of recurrent high-grade gliomas and the systemic BCNU exposure with increasing doses of interstitial BCNU. PATIENTS AND METHODS: Forty-four adults underwent tumor debulking and polymer placement. Six patients per dose level were studied using polymers with 6.5%, 10%, 14.5%, 20%, and 28% BCNU by weight. Toxicities were assessed 1 month after implantation by a safety monitoring committee to determine whether subsequent escalations should occur. Nine additional patients were studied at the MTD to confirm safety. BCNU blood levels were obtained before and after polymer implantation. RESULTS: No dose-limiting toxicities were identified at the 6.5%, 10%, or 14.5% dose levels, although difficulties with wound healing, seizures, and brain edema were noted. At the 20% dose, these effects seemed more prominent, and six additional patients were treated at this dose and tolerated treatment well. Three of four patients receiving the 28% polymers developed severe brain edema and seizures, and accrual to this cohort was stopped. Nine additional patients received 20% polymer, confirming this as the MTD. Maximum BCNU plasma concentrations with the 20% loaded polymers were 27 ng/mL. Overall median survival was 251 days. CONCLUSION: The MTD of BCNU delivered in polymer to the surgical cavity is 20%. This polymer provides five times more BCNU than standard commercially available BCNU polymers and results in minimal systemic BCNU exposure. Additional studies are needed to establish the efficacy of high-dose BCNU polymers. PMID- 12721264 TI - Phase I study of an oral formulation of ZD9331 administered daily for 28 days. AB - PURPOSE: To define the maximum-tolerated dose and dose-limiting toxicities (DLTs) of an oral formulation of ZD9331, a novel thymidylate synthase inhibitor that is not a substrate for folylpolyglutamate synthase. PATIENTS AND METHODS: Patients had Cancer and Leukemia Group B performance status < or = 2 and refractory solid tumors. Initially, patients received ZD9331 daily for 2 weeks, with the duration of treatment escalated to a maximum of 4 weeks, followed by a 2-week rest period. Once the maximum-tolerated duration of treatment was determined, the dose of ZD9331 was increased until DLT occurred. RESULTS: Fifty-five patients were enrolled at eight dose levels. The DLTs were thrombocytopenia and neutropenia. At 3 mg/d, two of 19 patients developed DLT; one patient had grade 3 thrombocytopenia and grade 4 neutropenia, and the other patient had grade 3 thrombocytopenia only. Anemia was common, with a median hemoglobin nadir of 75% of baseline, before recovery or transfusion. The apparent oral clearance of ZD9931 was 11.6 +/- 6.3 mL/min. Dose-limiting myelosuppression was associated with both an increased 24-hour ZD9931 concentration and blood urea nitrogen. CONCLUSION: The recommended phase II dose on this schedule is 3 mg/d for 4 weeks, followed by a 2-week rest period. ZD9331 seems to have a manageable toxicity profile, although it should be used with caution in patients with renal impairment. PMID- 12721265 TI - Phase I trial and pharmacokinetic study of BMS-247550, an epothilone B analog, administered intravenously on a daily schedule for five days. AB - PURPOSE: The epothilones are a novel class of nontaxane microtubule-stabilizing agents. BMS-247550 is a semisynthetic analog of the natural product epothilone B. We conducted a phase I study administering BMS-247550 as a 1-hour intravenous infusion daily for 5 consecutive days every 21 days. PATIENTS AND METHODS: Twenty one patients received BMS-247550 without filgrastim in the first cycle. An additional six patients were enrolled at a starting dose of 8 mg/m2/d with filgrastim support. Twenty-one of the 27 patients had received prior paclitaxel, docetaxel, or both. RESULTS: One hundred seven cycles were administered to 27 patients. The maximum-tolerated dose was 6 mg/m2 of BMS-247550 administered as a 1-hour intravenous infusion daily for 5 consecutive days every 21 days. Dose limiting toxicity at a dose of 8 mg/m2/d was neutropenia with or without filgrastim support. Nonhematologic grade 3 toxicities included fatigue (seven cycles), stomatitis (two cycles), and anorexia (one cycle). The mean terminal half-life of BMS-247550 was 16.8 +/- 6.0 hours, the volume of distribution at steady-state was 798 +/- 375 L, and the clearance was 712 +/- 247 mL/min. Objective responses were observed in patients with breast, cervical, and basal cell cancer. Reductions in CA-125 levels were noted in patients with ovarian cancer. CONCLUSION: The recommended phase II dose of BMS-247550 on the daily schedule for 5 days is 6 mg/m2/d. Neutropenia was dose limiting, but higher doses were tolerated by a large fraction of patients with filgrastim support. Peripheral neuropathy was mild, even after multiple cycles of therapy, and was not dose limiting. PMID- 12721266 TI - Monoclonal antibody therapy of chronic lymphocytic leukemia. AB - Chemotherapeutic approaches during the last decade have failed to result in major advances in the outcome of patients with chronic lymphocytic leukemia (CLL). The recent availability of an increasing number of active monoclonal antibodies, immunotoxins, and radioimmunoconjugates (RICs) has stimulated considerable interest in clinical research in CLL. Alemtuzumab was the first antibody approved for CLL on the basis of responses in one third of patients with advanced disease. However, infusion reactions and immunosuppression with opportunistic infections present a challenge that may be overcome with altered schedules and routes of administration. Rituximab has limited activity as a single agent in patients relapsed or refractory after prior chemotherapy; however, response rates seem to be higher in previously untreated patients. More importantly, combinations with chemotherapy drugs such as fludarabine are showing promise in early trials. Newer antibodies in development as single agents and in combinations include apolizumab (Hu1D10), a humanized antibody against an epitope of HLA-DR, and IDEC-152, a primatized anti-CD23 antibody. BL22, an immunotoxin with impressive activity in hairy cell leukemia, is in phase II trials in CLL as well. The safe use of RICs is complicated by the elevated peripheral blood B-cell count, and the extent of bone marrow involvement in CLL; studies will explore the use of agents to eliminate malignant cells from the bone marrow before RIC therapy. It is hoped that the rational development of combinations of the various promising antibodies with chemotherapy and each other will lead to more effective approaches for patients with CLL. PMID- 12721268 TI - Hematologic malignancies with extramedullary spread of disease. Case 1. Multiple myeloma with extramedullary involvement of the pleura and testes. PMID- 12721267 TI - Tobacco dependence: why should an oncologist care? PMID- 12721270 TI - Hematologic malignancies with extramedullary spread of disease. Case 3. Extra nodal Hodgkin's disease presenting as rapidly progressive liver failure. PMID- 12721269 TI - Hematologic malignancies with extramedullary spread of disease. Case 2. Hepatosplenic T-cell lymphoma. PMID- 12721271 TI - Effectiveness of adjuvant fluorouracil in elderly colon cancer patients: the internal and external validity of nonrandomized research design. PMID- 12721275 TI - Surrogate outcomes in quality-of-life research: where will we end up? PMID- 12721273 TI - Postoperative breast irradiation: new trials needed in older patients. PMID- 12721277 TI - Partnering between cancer centers: the American College of Surgeons, National Cancer Institute, American Cancer Society, Centers of Disease Control, American Society of Clinical Oncology, Society of Surgical Oncology, American Society of Therapeutic Radiation and Oncology, and Association of Community Cancer Centers are needed to stimulate improved clinical cancer care. PMID- 12721279 TI - Acute hypokalemic tetraparesis induced by intravenous methotrexate. PMID- 12721280 TI - American Society of Clinical Oncology: background for update of conflict of interest policy. PMID- 12721281 TI - American Society of Clinical Oncology policy statement: oversight of clinical research. AB - Well-publicized lapses in the review or implementation of clinical research studies have raised public questions about the integrity of the clinical research process. Public trust in the integrity of research is critical not only for funding and participation in clinical trials but also for confidence in the treatments that result from the trials. The questions raised by these unfortunate cases pose an important opportunity to reassess the clinical trials oversight system to ensure the integrity of clinical research and the safety of those who enroll in clinical trials. Since its inception, the American Society of Clinical Oncology (ASCO) has worked for the advancement of cancer treatments through clinical research and to help patients gain prompt access to scientifically excellent and ethically unimpeachable clinical trials. As an extension of its mission, ASCO is affirming with this policy statement the critical importance of a robust review and oversight system to ensure that clinical trials participants give fully informed consent and that their safety is a top priority. Ensuring the integrity of research cannot be stressed enough because of its seminal connection to the advancement of clinical cancer treatment. The overall goal of this policy is to enhance public trust in the cancer clinical trials process. To achieve this, the following elements are essential: 1. Ensure safety precautions for clinical trial participants and their fully informed consent. 2. Ensure the validity and integrity of scientific research. 3. Enhance the educational training of clinical scientists and research staff to ensure the highest standards of research conduct. 4. Promote accountability and responsibility among all those involved in clinical research (not just those serving on institutional review boards [IRBs], but also institutional officials, researchers, sponsors, and participants) and ensure support for an effective oversight process. 5. Enhance the professional and public understanding of clinical research oversight. 6. Enhance the efficiency and cost-effectiveness of the clinical research oversight system. This policy statement makes recommendations in several areas that serve as principles to support an improved system of oversight for clinical research. ASCO will work with all parties involved in the clinical research system to develop the steps necessary to implement these recommendations. Centralized Trial Review: A large percentage of oncology clinical trials are coordinated through the National Cancer Institute's (NCI) system of cooperative groups, which already incorporates centralized scientific review. As such, there is a tremendous opportunity to employ a centralized mechanism to provide ethical review by highly trained IRB members, allowing local IRBs to take advantage of the financial and time efficiencies that central review provides. Centralized review boards (CRBs) would also contribute consistency and efficiency to the process. Once successfully completed, the review would represent an approval to open the protocol at all of the institutions that have subscribed to the centralized review system. Local IRBs would be able to devote time usually spent on initial review to ongoing monitoring of the trial taking place at their institution. Considering the enormous size and complexity of the clinical research enterprise, ASCO envisions multiple CRBs, which could be distributed as regional review boards. Central review will use a single protocol and consent form, and monitor and evaluate adverse events (AEs) on a global basis, eliminating many of the time-consuming steps for the local IRB. Global monitoring and assessment of AEs has real potential to enhance trial participants' safety by giving local institutions more information on the overall trial and enabling them to devote more time to ongoing review of the trial onsite. Use of a CRB also has real potential to reduce the costs of clinical trial oversight by allowing local IRBs to eliminate the costs of initial review. These efficiencies will likely lead to institutions redirecting funds toward monitoring ongoing trials. Although a CRB has potential to improve the efficiency of the process, a CRB could also have tremendous ability to delay valuable trials. Checks and balances must be included in the newly devised system to ensure timely review and appeals of CRB actions. ASCO proposes the advent of a new pilot program for centralizing review of clinical trials. It requires clear engagement of all stakeholders in planning the experiment, clear articulation of the goals, and assurance of federal regulatory protection for institutions choosing to participate. If successful, this CRB pilot project could be expanded to include multi-institutional industry sponsored research. Education and Training:Education and training are critical to the ultimate success of an improved oversight system. All members of the research team should receive comprehensive education on conducting scientifically and ethically valid clinical research. The curriculum should also include information on the prevailing local and federal regulations that pertain to the clinical trials process. IRB members should also receive ongoing education and training in the review of clinical research protocols. IRB training should pay particular attention to nonscientific members to give them the tools necessary to speak on behalf of research participants. ASCO should develop a curriculum that focuses on the proper conduct of human research and emphasize ethically sound clinical research in the context of its Annual Meeting. Informed Consent: Investigators and review boards have specific roles to play in ensuring the education of trial participants through the informed consent process, both when they are considering trial enrollment and as they participate in the trial. Review boards and investigators should focus primarily on the informed consent process, rather than the informed consent documents. Federal Oversight: The federal government has an important role to play in the oversight of clinical research. This role should be expanded to cover all research, not just that which is funded by the federal government or conducted with the oversight of the Food and Drug Administration (FDA). The Department of Health and Human Services (HHS) Office for Human Research Protections (OHRP) and the FDA should provide clear regulatory support and guidance for local institutions that choose to employ a CRB. In the case of the pilot CRB discussed in this policy statement, it should serve as the preferred option for the cancer cooperative group clinical trials. Ideally, the federal government should unify and streamline its regulations for the oversight of clinical research. Resources Supporting Clinical Research Infrastructure:An effective oversight process demands the highest quality scientific and ethical review and onsite monitoring of the safety of trial participants. This can only be accomplished by the involvement of an experienced IRB that receives funding, resources, and institutional support enabling it to fulfill its mandate. CONFLICT OF INTEREST: Critical to the integrity of research is the absence of bias in the process. ASCO strongly recommends the adoption of standards for the identification, management, and, where appropriate, elimination of conflicts of interests, whether they are actual, potential, or apparent. PMID- 12721282 TI - American Society of Clinical Oncology: revised conflict of interest policy. PMID- 12721283 TI - TIRP, a novel Toll/interleukin-1 receptor (TIR) domain-containing adapter protein involved in TIR signaling. AB - The Toll/interleukin-1 receptor (TIR) family members play important roles in host defense. These receptors signal through TIR domain-containing adapter proteins. In this report, we identified a novel TIR domain-containing adapter protein designated as TIRP. Co-immunoprecipitation experiments suggest that TIRP is associated with IL-1 receptors. TIRP also interacts with kinase-inactive mutants of IRAK and IRAK-4, IRAK-2, IRAK-M, and TRAF6. Overexpression of TIRP activates NF-kappaB and potentiates IL-1 receptor-mediated NF-kappaB activation. A dominant negative mutant of TIRP inhibits IL-1- but not tumor necrosis factor-triggered NF kappaB activation. Moreover, TIRP-mediated NF-kappaB activation is inhibited by dominant negative mutants of IRAK, IRAK-2, TRAF6, and IKKbeta. Our findings suggest that TIRP is involved in IL-1-triggered NF-kappaB activation and functions upstream of IRAK, IRAK-2, TRAF6, and IKKbeta PMID- 12721284 TI - The C terminus (amino acids 75-94) and the linker region (amino acids 42-54) of the Ca2+-binding protein S100A1 differentially enhance sarcoplasmic Ca2+ release in murine skinned skeletal muscle fibers. AB - S100A1, a Ca2+-binding protein of the EF-hand type, is most highly expressed in striated muscle and has previously been shown to interact with the skeletal muscle sarcoplasmic reticulum (SR) Ca2+ release channel/ryanodine receptor (RyR1) isoform. However, it was unclear whether S100A1/RyR1 interaction could modulate SR Ca2+ handling and contractile properties in skeletal muscle fibers. Since S100A1 protein is differentially expressed in fast- and slow-twitch skeletal muscle, we used saponin-skinned murine Musculus extensor digitorum longus (EDL) and Musculus soleus (Soleus) fibers to assess the impact of S100A1 protein on SR Ca2+ release and isometric twitch force in functionally intact permeabilized muscle fibers. S100A1 equally enhanced caffeine-induced SR Ca2+ release and Ca2+ induced isometric force transients in both muscle preparations in a dose dependent manner. Introducing a synthetic S100A1 peptide model (devoid of EF-hand Ca2+-binding sites) allowed identification of the S100A1 C terminus (amino acids 75-94) and hinge region (amino acids 42-54) to differentially enhance SR Ca2+ release with a nearly 3-fold higher activity of the C terminus. These effects were exclusively based on enhanced SR Ca2+ release as S100A1 influenced neither SR Ca2+ uptake nor myofilament Ca2+ sensitivity/cooperativity in our experimental setting. In conclusion, our study shows for the first time that S100A1 augments contractile performance both of fast- and slow-twitch skeletal muscle fibers based on enhanced SR Ca2+ efflux at least mediated by the C terminus of S100A1 protein. Thus, our data suggest that S100A1 may serve as an endogenous enhancer of SR Ca2+ release and might therefore be of physiological relevance in the process of excitation-contraction coupling in skeletal muscle. PMID- 12721285 TI - Identification of regulatory domains in ADP-ribosyltransferase-1 that determine transferase and NAD glycohydrolase activities. AB - Mono-ADP-ribosyltransferases (ART1-7) transfer ADP-ribose from NAD+ to proteins (transferase activity) or water (NAD glycohydrolase activity). The mature proteins contain two domains, an alpha-helical amino terminus and a beta-sheet rich carboxyl terminus. A basic region in the carboxyl termini is encoded in a separate exon in ART1 and ART5. Structural motifs are conserved among ART molecules. Successive amino- or carboxyl-terminal truncations of ART1, an arginine-specific transferase, identified regions that regulated transferase and NAD glycohydrolase activities. In mouse ART1, amino acids 24-38 (ART-specific extension) were needed to inhibit both activities; amino acids 39-45 (common ART coil) were required for both. Successive truncations of the alpha-helical region reduced transferase and NAD glycohydrolase activities; however, truncation to residue 106 enhanced both. Removal of the carboxyl-terminal basic domain decreased transferase, but enhanced NAD glycohydrolase, activity. Thus, amino- and carboxyl-terminal regions of ART1 are required for transferase activity. The enhanced glycohydrolase activity of the shorter mutants indicates that sequences, which are not part of the NAD binding, core catalytic site, exert structural constraints, modulating substrate specificity and catalytic activity. These functional domains, defined by discrete exons or structural motifs, are found in ART1 and other ARTs, consistent with conservation of structure and function across the ART family. PMID- 12721286 TI - A novel RNA polymerase II C-terminal domain phosphatase that preferentially dephosphorylates serine 5. AB - The transcription and processing of pre-mRNA in eukaryotic cells are regulated in part by reversible phosphorylation of the C-terminal domain of the largest RNA polymerase (RNAP) II subunit. The CTD phosphatase, FCP1, catalyzes the dephosphorylation of RNAP II and is thought to play a major role in polymerase recycling. This study describes a family of small CTD phosphatases (SCPs) that preferentially catalyze the dephosphorylation of Ser5 within the consensus repeat. The preferred substrate for SCP1 is RNAP II phosphorylated by TFIIH. Like FCP1, the activity of SCP1 is enhanced by the RAP74 subunit of TFIIF. Expression of SCP1 inhibits activated transcription from a number of promoters, whereas a phosphatase-inactive mutant of SCP1 enhances transcription. Accordingly, SCP1 may play a role in the regulation of gene expression, possibly by controlling the transition from initiation/capping to processive transcript elongation. PMID- 12721287 TI - Distinct endosomal compartments in early trafficking of low density lipoprotein derived cholesterol. AB - We previously studied the early trafficking of low density lipoprotein (LDL) derived cholesterol in mutant Chinese hamster ovary cells defective in Niemann Pick type C1 (NPC1) using cyclodextrin (CD) to monitor the arrival of cholesterol from the cell interior to the plasma membrane (PM) (Cruz, J. C., Sugii, S., Yu, C., and Chang, T.-Y. (2000) J. Biol. Chem. 275, 4013-4021). We found that newly hydrolyzed cholesterol derived from LDL first appears in certain CD-accessible pool(s), which we assumed to be the PM, before accumulating in the late endosome/lysosome, where NPC1 resides. To determine the identity of the early CD accessible pool(s), in this study, we performed additional experiments, including the use of revised CD incubation protocols. We found that prolonged incubation with CD (>30 min) caused cholesterol in internal membrane compartment(s) to redistribute to the PM, where it became accessible to CD. In contrast, a short incubation with CD (5-10 min) did not cause such an effect. We also show that one of the early compartments contains acid lipase (AL), the enzyme required for liberating cholesterol from cholesteryl ester in LDL. Biochemical and microscopic evidence indicates that most of the AL is present in endocytic compartment(s) distinct from the late endosome/lysosome. Our results suggest that cholesterol is liberated from LDL cholesteryl ester in the hydrolytic compartment containing AL and then moves to the NPC1-containing late endosome/lysosome before reaching the PM or the endoplasmic reticulum. PMID- 12721288 TI - BCL-2 and BCL-XL restrict lineage choice during hematopoietic differentiation. AB - Differentiation of hematopoietic cells from multipotential progenitors is regulated by multiple growth factors and cytokines. A prominent feature of these soluble factors is promotion of cell survival, in part mediated by expression of either of the anti-apoptotic proteins, BCL-2 and BCL-XL. The complex expression pattern of these frequently redundant survival factors during hematopoiesis may indicate a role in lineage determination. To investigate the latter possibility, we analyzed factor-dependent cell-Patersen (FDCP)-Mix multipotent progenitor cells in which we stably expressed BCL-2 or BCL-XL. Each factor maintained complete survival of interleukin-3 (IL-3)-deprived FDCP-Mix cells but, unexpectedly, directed FDCP-Mix cells along restricted and divergent differentiation pathways. Thus, IL-3-deprived FDCP-Mix BCL-2 cells differentiated exclusively to granulocytes and monocytes/macrophages, whereas FDCP-Mix BCL-XL cells became erythroid. FDCP-Mix BCL-2 cells grown in IL-3 were distinguished from FDCP-Mix and FDCP-Mix BCL-XL cells by a striking reduction in cellular levels of Raf-1 protein. Replacement of the BCL-2 BH4 domain with the related BCL XL BH4 sequence resulted in a switch of FDCP-Mix BCL-2 cells to erythroid fate accompanied by persistence of Raf-1 protein expression. Moreover, enforced expression of Raf-1 redirected FDCP-Mix BCL-2 cells to an erythroid fate, and prohibited generation of myeloid cells. These results identify novel roles for BCL-2 and BCL-XL in cell fate decisions beyond cell survival. These effects are associated with differential regulation of Raf-1 expression, perhaps involving the previously identified interaction between BCL-2-BH4 and the catalytic domain of Raf-1. PMID- 12721289 TI - Involvement of integrins in osmosensing and signaling toward autophagic proteolysis in rat liver. AB - Inhibition of autophagic proteolysis by hypoosmotic or amino acid-induced hepatocyte swelling requires osmosignaling toward p38MAPK; however, the upstream osmosensing and signaling events are unknown. These were studied in the intact perfused rat liver with a preserved in situ environment of hepatocytes. It was found that hypoosmotic hepatocyte swelling led to an activation of Src (but not FAK), Erks, and p38MAPK, which was prevented by the integrin inhibitory hexapeptide GRGDSP, but not its inactive analogue GRGESP. Src inhibition by PP-2 prevented hypoosmotic MAP kinase activation, indicating that the integrin/Src system is located upstream in the osmosignaling toward p38MAPK and Erks. Inhibition of the integrin/Src system by the RGD motif-containing peptide or PP-2 also prevented the inhibition of proteolysis and the decrease in autophagic vacuole volume, which is otherwise observed in response to hypoosmotic or glutamine/glycine-induced hepatocyte swelling. These inhibitors, however, did not affect swelling-independent proteolysis inhibition by phenylalanine. In line with a role of p38MAPK in triggering the volume regulatory decrease (RVD), PP-2 and the RGD peptide blunted RVD in response to hypoosmotic cell swelling. The data identify integrins and Src as upstream events in the osmosignaling toward MAP kinases, proteolysis, and RVD. They further point to a role of integrins as osmo- and mechanosensors in the intact liver, which may provide a link between cell volume and cell function. PMID- 12721290 TI - Close homolog of L1 is an enhancer of integrin-mediated cell migration. AB - Close homolog of L1 (CHL1) is a member of the L1 family of cell adhesion molecules expressed by subpopulations of neurons and glia in the central and peripheral nervous system. It promotes neurite outgrowth and neuronal survival in vitro. This study describes a novel function for CHL1 in potentiating integrin dependent cell migration toward extracellular matrix proteins. Expression of CHL1 in HEK293 cells stimulated their haptotactic migration toward collagen I, fibronectin, laminin, and vitronectin substrates in Transwell assays. CHL1 potentiated cell migration to collagen I was dependent on alpha1beta1 and alpha2beta1 integrins, as shown with function blocking antibodies. Potentiated migration relied on the early integrin signaling intermediates c-Src, phosphatidylinositol 3-kinase, and mitogen-activated protein kinase. Enhancement of migration was disrupted by mutation of a potential integrin interaction motif Asp-Gly-Glu-Ala (DGEA) in the sixth immunoglobulin domain of CHL1, suggesting that CHL1 functionally interacts with beta1 integrins through this domain. CHL1 was shown to associate with beta1 integrins on the cell surface by antibody induced co-capping. Through a cytoplasmic domain sequence containing a conserved tyrosine residue (Phe-Ile-Gly-Ala-Tyr), CHL1 recruited the actin cytoskeletal adapter protein ankyrin to the plasma membrane, and this sequence was necessary for promoting integrin-dependent migration to extracellular matrix proteins. These results support a role for CHL1 in integrin-dependent cell migration that may be physiologically important in regulating cell migration in nerve regeneration and cortical development. PMID- 12721291 TI - BCL-2 selectively interacts with the BID-induced open conformer of BAK, inhibiting BAK auto-oligomerization. AB - Caspase-8 cleaves BID to tBID, which targets mitochondria and induces oligomerization of BAX and BAK within the outer membrane, resulting in release of cytochrome c from the organelle. Here, we have initiated these steps in isolated mitochondria derived from control and BCL-2-overexpressing cells using synthetic BH3 peptides and subsequently analyzed the BCL members by chemical cross-linking. The results show that the BH3 domain of BID interacts with and induces an "open" conformation of BAK, exposing the BAK N terminus. This open (activated) conformer of BAK potently induces oligomerization of non-activated ("closed") conformers, causing a cascade of BAK auto-oligomerization. Induction of the open conformation of BAK occurs even in the presence of excess BCL-2, but BCL-2 selectively interacts with this open conformer and blocks BAK oligomerization and cytochrome c release, dependent on the ratio of BID BH3 and BCL-2. This mechanism of inhibition by BCL-2 also occurs in intact cells stimulated with Fas or expressing tBID. Although BID BH3 interacts with both BCL-2 and BAK, the results indicate that when BCL-2 is in excess it can sequester the BID BH3-induced activated conformer of BAK, effectively blocking downstream events. This model suggests that the primary mechanism for BCL-2 blockade targets activated BAK rather than sequestering tBID. PMID- 12721292 TI - A combinatorial G protein-coupled receptor reconstitution system on budded baculovirus. Evidence for Galpha and Galphao coupling to a human leukotriene B4 receptor. AB - To investigate the coupling selectivity of G proteins and G protein-coupled receptors (GPCRs), we developed a reconstitution system made up of GPCR and heterotrimeric G proteins on extracellular baculovirus particles (budded virus (BV)). BV released from Sf9 cells infected with a recombinant baculovirus coding for human leukotriene B4 receptor (BLT1) cDNA exhibited a high level of BLT1 expression (27.3 pmol/mg of protein) and specific [3H]leukotriene B4 binding activity (Kd = 3.67 nm). The apparent low affinity of the expressed BLT1 is thought to be due to relative non-availability of the Galphai isoform, which couples to BLT1, in BV. Co-infection of heterotrimeric G protein recombinant viruses led to co-expression of BLT1 and G protein subunits on BV. A guanosine-5' (beta,gamma-imido)triphosphate-sensitive, high affinity ligand binding was observed in the BLT1 BV co-expressing Galphai1beta1gamma2 (Kd = 0.17 nm). A relatively large amount of high affinity receptor protein was recovered in the co expressing BV fraction (6.81 pmol/mg of protein). A combination of BLT1 and Galphai1 without Gbeta1gamma2 did not exhibit high affinity ligand binding on BV, indicating the low background environment for the GPCR-G protein coupling in this BV reconstitution system. To test other G proteins for coupling, various Galpha subunits were combinatorially expressed in BV with BLT1 and Gbeta1gamma2. The BLT1 BV co-expressing GalphaoAbeta1gamma2 exhibited a comparably high affinity ligand binding as well as ligand-stimulated guanosine 5'-3-O-(thio)triphosphate binding to Galphai1beta1gamma2. Co-expression of other Galpha isoforms such as Galphas, Galpha11, Galpha14, Galpha16, Galpha12, or Galpha13 did not exhibit any significant effects on ligand binding affinity in this system. These results reveal that BLT1 and coupled trimeric G proteins were functionally reconstituted on BV and that Galphao as well as Galphai couples to BLT1. This expression system should prove highly useful for pharmacological characterization, biosensor chip applications, and also drug discovery directed at highly important targets of the membrane receptor proteins. PMID- 12721293 TI - Uptake and metabolism of low density lipoproteins with elevated ceramide content by human microvascular endothelial cells: implications for the regulation of apoptosis. AB - Ceramide is a bioactive molecule involved in cellular responses to stress and inflammation. The major pathway for ceramide accumulation is via agonist-induced activation of cellular sphingomyelinases. It has also been shown that the ceramide level in circulating low density lipoprotein (LDL) increases during systemic inflammation, hence it is of importance to understand whether LDL derived ceramide also contributes to the cellular ceramide homeostasis and affects cell functions. This article provides evidence of uptake of ceramide enriched LDL by human microvascular endothelial cells in a receptor-mediated fashion. This uptake can be competed by native LDL, indicating that the LDL binding receptor may be involved. Following uptake, part of the LDL-derived ceramide is converted to sphingosine, but more importantly, part of it accumulates inside the cells (approximately 1.44 nmol/mg of cell protein). This accumulation of ceramide correlates with an increased incidence of apoptosis. The addition of tumor necrosis factor-alpha further enhances the accumulation of LDL derived ceramide and the rate of apoptosis. In contrast, inhibitors of receptor mediated endocytosis block both, the accumulation of LDL-derived ceramide and the concurrent increase in apoptosis. We also show that LDL-delivered ceramide is a more efficient inducer of apoptosis as compared with ethanol-delivered ceramide, the same apoptotic effect being achieved by substantially smaller increases in intracellular ceramide. Taken together, the presented data indicate that increases in lipoprotein ceramide concentration may result in changes in the bioactive properties of circulating lipoproteins such as the enhanced ability to induce apoptosis in the microvascular endothelium. PMID- 12721294 TI - Purification, kinetic characterization, and molecular cloning of a novel enzyme ecdysteroid-phosphate phosphatase. AB - From eggs of the silkworm Bombyx mori, we isolated a novel enzyme that is involved in the conversion of physiologically inactive conjugated ecdysteroids, such as ecdysone 22-phosphate and 20-hydroxyecdysone 22-phosphate, to active free ecdysteroids. This enzyme, called ecdysteroid-phosphate phosphatase (EPPase), was located in the cytosol fraction and differed from nonspecific lysosomal acid phosphatases in various enzymic properties. EPPase was purified about 3,000-fold to homogeneity by seven steps of column chromatography. The cDNA clone encoding EPPase was isolated by reverse transcription polymerase chain reaction using degenerate primers on the basis of the partial amino acid sequence obtained from purified EPPase and by subsequent 3'- and 5'-rapid amplification of cDNA ends. The full-length cDNA of EPPase was found to be composed of 1620 bp with an open reading frame encoding a protein of 331 amino acid residues. A data base search showed that there was no functional protein with the amino acid sequence identical to that of EPPase. Northern blot analysis revealed that EPPase mRNA was expressed predominantly during gastrulation and organogenesis in nondiapause eggs but was not detected in diapause eggs whose development was arrested at the late gastrula stage. In nondiapause eggs, the developmental changes in the expression pattern of EPPase mRNA corresponded closely to changes in the enzyme activity and in the amounts of free ecdysteroids in eggs. PMID- 12721295 TI - Dentin sialophosphoprotein knockout mouse teeth display widened predentin zone and develop defective dentin mineralization similar to human dentinogenesis imperfecta type III. AB - Dentin sialophosphoprotein (Dspp) is mainly expressed in teeth by the odontoblasts and preameloblasts. The Dspp mRNA is translated into a single protein, Dspp, and cleaved into two peptides, dentin sialoprotein and dentin phosphoprotein, that are localized within the dentin matrix. Recently, mutations in this gene were identified in human dentinogenesis imperfecta II (Online Mendelian Inheritance in Man (OMIM) accession number 125490) and in dentin dysplasia II (OMIM accession number 125420) syndromes. Herein, we report the generation of Dspp-null mice that develop tooth defects similar to human dentinogenesis imperfecta III with enlarged pulp chambers, increased width of predentin zone, hypomineralization, and pulp exposure. Electron microscopy revealed an irregular mineralization front and a lack of calcospherites coalescence in the dentin. Interestingly, the levels of biglycan and decorin, small leucine-rich proteoglycans, were increased in the widened predentin zone and in void spaces among the calcospherites in the dentin of null teeth. These enhanced levels correlate well with the defective regions in mineralization and further indicate that these molecules may adversely affect the dentin mineralization process by interfering with coalescence of calcospherites. Overall, our results identify a crucial role for Dspp in orchestrating the events essential during dentin mineralization, including potential regulation of proteoglycan levels. PMID- 12721296 TI - The protein tyrosine phosphatase SHP-2 regulates interleukin-1-induced ERK activation in fibroblasts. AB - Focal adhesion complexes are actin-rich, cytoskeletal structures that mediate cell adhesion to the substratum and also selectively regulate signal transduction pathways required for interleukin (IL)-1beta signaling to the MAP kinase, ERK. IL 1-induced ERK activation is markedly diminished in fibroblasts deprived of focal adhesions whereas activation of p38 and JNK is unaffected. While IL-1 signaling is known to involve the activity of protein and lipid kinases including MAP kinases, FAK, and PI3K, little is known about the role of phosphatases in the regulation of IL-1 signal generation and attenuation. Here we demonstrate that SHP-2, a protein tyrosine phosphatase present in focal adhesions, modulates IL-1 induced ERK activation and the transient actin stress fiber disorganization that occurs following IL-1 treatment in human gingival fibroblasts. Using a combination of immunoblotting, immunoprecipitation, and immunostaining we show that SHP-2 is present in nascent focal adhesions and undergoes phosphorylation on tyrosine 542 in response to IL-1 stimulation. Blocking anti-SHP-2 antibodies, electoporated into the cytosol of fibroblasts, inhibited IL-1-induced ERK activation, actin filament assembly, and cell contraction, indicating a role for SHP-2 in these processes. In summary, our data indicate that SHP-2, a focal adhesion-associated protein, participates in IL-1-induced ERK activation likely via an adaptor function. PMID- 12721297 TI - Structural analysis of the mitotic regulator hPin1 in solution: insights into domain architecture and substrate binding. AB - The peptidyl-prolyl cis/trans isomerase hPin1 is a phosphorylation-dependent regulatory enzyme whose substrates are proteins involved in regulation of cell cycle, transcription, Alzheimer's disease, and cancer pathogenesis. We have determined the solution structure of the two domain protein hPin1-(1-163) and its separately expressed PPIase domain (50-163) (hPin1PPIase) with an root mean square deviation of <0.5 A over backbone atoms using NMR. Domain organization of hPin1 differs from that observed in structures solved by x-ray crystallography. Whereas PPIase and WW domain are tightly packed onto each other and share a common binding interface in crystals, our NMR-based data revealed only weak interaction of both domains at their interface in solution. Interaction between the two domains of full-length hPin1 is absent when the protein is dissected into the catalytic and the WW domain. It indicates that the flexible linker, connecting both domains, promotes binding. By evaluation of NOESY spectra we can show that the alpha1/beta1 loop, which was proposed to undergo a large conformational rearrangement in the absence of sulfate and an Ala-Pro peptide, remained in the closed conformation under these conditions. Dissociation constants of 0.4 and 2.0 mm for sulfate and phosphate ions were measured at 12 degrees C by fluorescence spectroscopy. Binding of sulfate prevents hPin1 aggregation and changes surface charges across the active center and around the reactive and catalytically essential Cys113. In the absence of sulfate and/or reducing agent this residue seems to promote aggregation, as observed in hPin1 solutions in vitro. PMID- 12721299 TI - Physical and functional interaction between protein kinase C delta and Fyn tyrosine kinase in human platelets. AB - An increasing number of tyrosine kinases have been shown to associate with isoforms of the protein kinase C (PKC) family. Here, we show evidence for physical and functional interaction between PKCdelta and the Src family kinase Fyn in human platelets activated by alboaggregin-A, a snake venom capable of activating both GPIb-V-IX and GPVI adhesion receptors. This interaction involves phosphorylation of PKCdelta on tyrosine and is specific in that other isoforms of PKC, PKCepsilon and lambda, which also become tyrosine-phosphorylated, do not interact with Fyn. In addition, PKCdelta does not interact with other platelet expressed tyrosine kinases Syk, Src, or Btk. Stimulation also leads to activation of both Fyn and PKCdelta and to serine phosphorylation of Fyn within a PKC consensus sequence. Alboaggregin-A-dependent activation of Fyn is blocked by bisindolylmaleimide I, suggesting a role for PKC isoforms in regulating Fyn activity. Platelet activation with alboaggregin-A induces translocation of the two kinases from cytoplasm to the plasma membrane of platelets, as observed by confocal immunofluorescence microscopy. Translocation of Fyn and PKCdelta are blocked by PP1 and bisindolylmaleimide I, showing a dependence upon Src and PKC kinase activities. Although PKC activity is required for translocation, it is not required for association between the two kinases, because this was not blocked by bisindolylmaleimide I. Rottlerin, which inhibited PKCdelta activity, did not block translocation of either PKCdelta or Fyn but potentiated platelet aggregation, 5-hydroxytryptamine secretion, and the calcium response induced by alboaggregin-A, indicating that this kinase plays a negative role in the control of these processes. PMID- 12721298 TI - Syntaxin 1A modulates the voltage-gated L-type calcium channel (Ca(v)1.2) in a cooperative manner. AB - Syntaxin 1A (Sx1A) modifies the activity of voltage-gated Ca2+ channels acting via the cytosolic and the two vicinal cysteines (271 and 272) at the transmembrane domain. Here we show that Sx1A modulates the Lc-type Ca2+ channel, Cav1.2, in a cooperative manner, and we explore whether channel clustering or the Sx1A homodimer is responsible for this activity. Sx1A formed homodimers but, when mutated at the two vicinal transmembrane domain cysteines, was unable to either dimerize or modify the channel activity suggesting disulfide bond formation. Moreover, applying global molecular dynamic search established a theoretical prospect of generating a disulfide bond between two Sx1A transmembrane helices. Nevertheless, Sx1A activity was not correlated with Sx1A homodimer. Application of a vicinal thiol reagent, phenylarsine oxide, abolished Sx1A action indicating the accessibility of Cys-271,272 thiols. Sx1A inhibition of channel activity was restored by phenylarsine oxide antidote, 2,3-dimercaptopropanol, consistent with thiol interaction of Sx1A. In addition, the supralinear mode of channel inhibition was correlated to the monomeric form of Sx1A and was apparent only when the three channel subunits alpha11.2/alpha2delta1/beta2a were present. This functional demonstration of cooperativity suggests that the three-subunit channel responds as a cluster, and Sx1A monomers associate with a dimer (or more) of a three-subunit Ca2+ channel. Consistent with channel cluster linked to Sx1A, a conformational change driven by membrane depolarization and Ca2+ entry would rapidly be transduced to the exocytotic machinery. As shown herein, the supralinear relationship between Sx1A and the voltage-gated Ca2+ channel within the cluster could convey the cooperativity that distinguishes the process of neurotransmitter release. PMID- 12721300 TI - Contribution of basic residues of the autolysis loop to the substrate and inhibitor specificity of factor IXa. AB - The autolysis loop (residues 143-154 in chymotrypsinogen numbering) plays a pivotal role in determining the macromolecular substrate and inhibitor specificity of coagulation proteases. This loop in factor IXa (FIXa) has 3 basic residues (Arg143, Lys147, and Arg150) whose contribution to the protease specificity of factor IXa has not been studied. Here, we substituted these residues individually with Ala in Gla-domainless forms of recombinant factor IX expressed in mammalian cells. All mutants exhibited normal amidolytic activities toward a FIXa-specific chromogenic substrate. However, Arg143 and Lys147 mutants showed a approximately 3- to 6-fold impairment in FX activation, whereas the Arg150 mutant activated factor X normally both in the absence and presence of factor VIIIa. By contrast, Arg143 and Lys147 mutants reacted normally with antithrombin (AT) in both the absence and presence of the cofactor, heparin. However, the reactivity of the Arg150 mutant with AT was impaired 6.6-fold in the absence of heparin and 33- to 70-fold in the presence of pentasaccharide and full length heparins. These results suggest that Arg143 and Lys147 of the autolysis loop are recognition sites for FX independent of factor VIIIa, and Arg150 is a specific recognition site for AT that can effectively interact with AT only if the serpin is in the heparin-activated conformation. PMID- 12721301 TI - Down-regulation of Myc as a potential target for growth arrest induced by human polynucleotide phosphorylase (hPNPaseold-35) in human melanoma cells. AB - Terminal differentiation and senescence share several common properties, including irreversible cessation of growth and changes in gene expression profiles. To identify molecules that converge in both processes, an overlapping pathway screening was employed that identified old-35, which is human polynucleotide phosphorylase (hPNPaseold-35), a 3',5'-exoribonuclease. We previously demonstrated that hPNPaseold-35 is a type I interferon-inducible gene that is also induced in senescent fibroblasts. In vitro RNA degradation assays confirmed its exoribonuclease properties, and overexpression of hPNPaseold-35 resulted in growth suppression in HO-1 human melanoma cells. The present study examined the molecular mechanism of the growth-arresting property of hPNPaseold 35. When overexpressed by means of a replication-incompetent adenoviral vector (Ad.hPNPaseold-35), hPNPaseold-35 inhibited cell growth in all cell lines tested. Analysis of cell cycle revealed that infection of HO-1 cells with Ad.hPNPaseold 35 resulted in arrest in the G1 phase and eventually apoptosis accompanied by marked reduction in the S phase. Infection with Ad.hPNPaseold-35 resulted in reduction in expression of the c-myc mRNA and Myc protein and modulated the expression of proteins regulating G1 checkpoint and apoptosis. In vitro mRNA degradation assays revealed that hPNPaseOLD-35 degraded c-myc mRNA. Overexpression of Myc partially but significantly protected HO-1 cells from Ad.hPNPaseold-35-induced growth arrest, indicating that Myc down-regulation might directly mediate the growth-inhibitory properties of Ad.hPNPaseold-35. Inhibition of hPNPaseold-35 by an antisense approach provided partial but significant protection against interferon-beta-mediated growth inhibition, thus demonstrating the biological significance of hPNPaseold-35 in interferon action. PMID- 12721302 TI - Regulation of canonical transient receptor potential (TRPC) channel function by diacylglycerol and protein kinase C. AB - The mechanism of receptor-induced activation of the ubiquitously expressed family of mammalian canonical transient receptor potential (TRPC) channels has been the focus of intense study. Primarily responding to phospholipase C (PLC)-coupled receptors, the channels are reported to receive modulatory input from diacylglycerol, endoplasmic reticulum inositol 1,4,5-trisphosphate receptors and Ca2+ stores. Analysis of TRPC5 channels transfected within DT40 B cells and deletion mutants thereof revealed efficient activation in response to PLC-beta or PLC-gamma activation, which was independent of inositol 1,4,5-trisphoshate receptors or the content of stores. In both HEK293 cells and DT40 cells, TRPC5 and TRPC3 channel responses to PLC activation were highly analogous, but only TRPC3 and not TRPC5 channels responded to the addition of the permeant diacylglycerol (DAG) analogue, 1-oleoyl-2-acetyl-sn-glycerol (OAG). However, OAG application or elevated endogenous DAG, resulting from either DAG lipase or DAG kinase inhibition, completely prevented TRPC5 or TRPC4 activation. This inhibitory action of DAG on TRPC5 and TRPC4 channels was clearly mediated by protein kinase C (PKC), in distinction to the stimulatory action of DAG on TRPC3, which is established to be PKC-independent. PKC activation totally blocked TRPC3 channel activation in response to OAG, and the activation was restored by PKC blockade. PKC inhibition resulted in decreased TRPC3 channel deactivation. Store operated Ca2+ entry in response to PLC-coupled receptor activation was substantially reduced by OAG or DAG-lipase inhibition in a PKC-dependent manner. However, store-operated Ca2+ entry in response to the pump blocker, thapsigargin, was unaffected by PKC. The results reveal that each TRPC subtype is strongly inhibited by DAG-induced PKC activation, reflecting a likely universal feedback control on TRPCs, and that DAG-mediated PKC-independent activation of TRPC channels is highly subtype-specific. The profound yet distinct control by PKC and DAG of the activation of TRPC channel subtypes is likely the basis of a spectrum of regulatory phenotypes of expressed TRPC channels. PMID- 12721303 TI - Calpains mediate p53 activation and neuronal death evoked by DNA damage. AB - DNA damage is an initiator of neuronal death implicated in neuropathological conditions such as stroke. Previous evidence has shown that apoptotic death of embryonic cortical neurons treated with the DNA damaging agent camptothecin is dependent upon the tumor suppressor p53, an upstream death mediator, and more distal death effectors such as caspases. We show here that the calcium-regulated cysteine proteases, calpains, are activated during DNA damage induced by camptothecin treatment. Moreover, calpain deficiency, calpastatin expression, or pharmacological calpain inhibitors prevent the death of embryonic cortical neurons, indicating the important role of calpain in DNA damage-induced death. Calpain inhibition also significantly reduced and delayed the induction of p53. Consistent with the actions of calpains upstream of p53 and the proximal nature of p53 death signaling, calpain inhibition inhibited cytochrome c release and DEVD-AFC cleavage activity. Taken together, our results indicate that calpains are a key mediator of p53 induction and consequent caspase-dependent neuronal death due to DNA damage. PMID- 12721304 TI - Identification and characterization of the human mus81-eme1 endonuclease. AB - The faithful and complete replication of DNA is necessary for the maintenance of genome stability. It is known, however, that replication forks stall at lesions in the DNA template and need to be processed so that replication restart can occur. In fission yeast, the Mus81-Eme1 endonuclease complex (Mus81-Mms4 in Saccharomyces cerevisiae) has been implicated in the processing of aberrant replication intermediates. In this report, we identify the human homolog of the Schizosaccharomyces pombe EME1 gene and have purified the human Mus81-Eme1 heterodimer. We show that Mus81-Eme1 is an endonuclease that exhibits a high specificity for synthetic replication fork structures and 3'-flaps in vitro. The nuclease cleaves Holliday junctions inefficiently ( approximately 75-fold less than flap or fork structures), although cleavage can be increased 6-fold by the presence of homologous sequences previously shown to permit base pair "breathing." We conclude that human Mus81-Eme1 is a flap/fork endonuclease that is likely to play a role in the processing of stalled replication fork intermediates. PMID- 12721305 TI - Electron transfer in cyanobacterial photosystem I: I. Physiological and spectroscopic characterization of site-directed mutants in a putative electron transfer pathway from A0 through A1 to FX. AB - The Photosystem I (PS I) reaction center contains two branches of nearly symmetric cofactors bound to the PsaA and PsaB heterodimer. From the x-ray crystal structure it is known that Trp697PsaA and Trp677PsaB are pi-stacked with the head group of the phylloquinones and are H-bonded to Ser692PsaA and Ser672PsaB, whereas Arg694PsaA and Arg674PsaB are involved in a H-bonded network of side groups that connects the binding environments of the phylloquinones and FX. The mutants W697FPsaA, W677FPsaB, S692CPsaA, S672CPsaB, R694APsaA, and R674APsaB were constructed and characterized. All mutants grew photoautotrophically, yet all showed diminished growth rates compared with the wild-type, especially at higher light intensities. EPR and electron nuclear double resonance (ENDOR) studies at both room temperature and in frozen solution showed that the PsaB mutants were virtually identical to the wild-type, whereas significant effects were observed in the PsaA mutants. Spin polarized transient EPR spectra of the P700+A1- radical pair show that none of the mutations causes a significant change in the orientation of the measured phylloquinone. Pulsed ENDOR spectra reveal that the W697FPsaA mutation leads to about a 5% increase in the hyperfine coupling of the methyl group on the phylloquinone ring, whereas the S692CPsaA mutation causes a similar decrease in this coupling. The changes in the methyl hyperfine coupling are also reflected in the transient EPR spectra of P700+A1- and the CW EPR spectra of photoaccumulated A1-. We conclude that: (i) the transient EPR spectra at room temperature are predominantly from radical pairs in the PsaA branch of cofactors; (ii) at low temperature the electron cycle involving P700 and A1 similarly occurs along the PsaA branch of cofactors; and (iii) mutation of amino acids in close contact with the PsaA side quinone leads to changes in the spin density distribution of the reduced quinone observed by EPR. PMID- 12721306 TI - Electron transfer in cyanobacterial photosystem I: II. Determination of forward electron transfer rates of site-directed mutants in a putative electron transfer pathway from A0 through A1 to FX. AB - The directionality of electron transfer in Photosystem I (PS I) is investigated using site-directed mutations in the phylloquinone (QK) and FX binding regions of Synnechocystis sp. PCC 6803. The kinetics of forward electron transfer from the secondary acceptor A1 (phylloquinone) were measured in mutants using time resolved optical difference spectroscopy and transient EPR spectroscopy. In whole cells and PS I complexes of the wild-type both techniques reveal a major, slow kinetic component of tau approximately 300 ns while optical data resolve an additional minor kinetic component of tau approximately 10 ns. Whole cells and PS I complexes from the W697FPsaA and S692CPsaA mutants show a significant slowing of the slow kinetic component, whereas the W677FPsaB and S672CPsaB mutants show a less significant slowing of the fast kinetic component. Transient EPR measurements at 260 K show that the slow phase is approximately 3 times slower than at room temperature. Simulations of the early time behavior of the spin polarization pattern of P700+A1-, in which the decay rate of the pattern is assumed to be negligibly small, reproduce the observed EPR spectra at 260 K during the first 100 ns following laser excitation. Thus any spin polarization from P700+FX- in this time window is very weak. From this it is concluded that the relative amplitude of the fast phase is negligible at 260 K or its rate is much less temperature-dependent than that of the slow component. Together, the results demonstrate that the slow kinetic phase results from electron transfer from QK-A to FX and that this accounts for at least 70% of the electrons. Although the assignment of the fast kinetic phase remains uncertain, it is not strongly temperature dependent and it represents a minor fraction of the electrons being transferred. All of the results point toward asymmetry in electron transfer, and indicate that forward transfer in cyanobacterial PS I is predominantly along the PsaA branch. PMID- 12721307 TI - The Saccharomyces cerevisiae COQ6 gene encodes a mitochondrial flavin-dependent monooxygenase required for coenzyme Q biosynthesis. AB - Coenzyme Q (Q) is a lipid that functions as an electron carrier in the mitochondrial respiratory chain in eukaryotes. There are eight complementation groups of Q-deficient Saccharomyces cerevisiae mutants, designated coq1-coq8. Here we have isolated the COQ6 gene by functional complementation and, in contrast to a previous report, find it is not an essential gene. coq6 mutants are unable to grow on nonfermentable carbon sources and do not synthesize Q but instead accumulate the Q biosynthetic intermediate 3-hexaprenyl-4-hydroxybenzoic acid. The Coq6 polypeptide is imported into the mitochondria in a membrane potential-dependent manner. Coq6p is a peripheral membrane protein that localizes to the matrix side of the inner mitochondrial membrane. Based on sequence homology to known proteins, we suggest that COQ6 encodes a flavin-dependent monooxygenase required for one or more steps in Q biosynthesis. PMID- 12721308 TI - Fas-associated death domain protein and caspase-8 are not recruited to the tumor necrosis factor receptor 1 signaling complex during tumor necrosis factor-induced apoptosis. AB - Death receptors are a subfamily of the tumor necrosis factor (TNF) receptor subfamily. They are characterized by a death domain (DD) motif within their intracellular domain, which is required for the induction of apoptosis. Fas associated death domain protein (FADD) is reported to be the universal adaptor used by death receptors to recruit and activate the initiator caspase-8. CD95, TNF-related apoptosis-inducing ligand (TRAIL-R1), and TRAIL-R2 bind FADD directly, whereas recruitment to TNF-R1 is indirect through another adaptor TNF receptor-associated death domain protein (TRADD). TRADD also binds two other adaptors receptor-interacting protein (RIP) and TNF-receptor-associated factor 2 (TRAF2), which are required for TNF-induced NF-kappaB and c-Jun N-terminal kinase activation, respectively. Analysis of the native TNF signaling complex revealed the recruitment of RIP, TRADD, and TRAF2 but not FADD or caspase-8. TNF failed to induce apoptosis in FADD- and caspase-8-deficient Jurkat cells, indicating that these apoptotic mediators were required for TNF-induced apoptosis. In an in vitro binding assay, the intracellular domain of TNF-R1 bound TRADD, RIP, and TRAF2 but did not bind FADD or caspase-8. Under the same conditions, the intracellular domain of both CD95 and TRAIL-R2 bound both FADD and caspase-8. Taken together these results suggest that apoptosis signaling by TNF is distinct from that induced by CD95 and TRAIL. Although caspase-8 and FADD are obligatory for TNF mediated apoptosis, they are not recruited to a TNF-induced membrane-bound receptor signaling complex as occurs during CD95 or TRAIL signaling, but instead must be activated elsewhere within the cell. PMID- 12721309 TI - Regulation of Bcl-xL expression by H2O2 in cardiac myocytes. AB - Oxidative stress promotes cardiac myocyte apoptosis through the mitochondrial death pathway. Since Bcl-2 family proteins are key regulators of apoptosis, we examined the effects of H2O2 on the expression of principal Bcl-2 family proteins (Bcl-2, Bcl-xL, Bax, Bad) in neonatal rat cardiac myocytes. Protein expression was assessed by immunoblotting. Bcl-2, Bax, and Bad were all down-regulated in myocytes exposed to 0.2 mm H2O2, a concentration that induces apoptosis. In contrast, although Bcl-xL levels initially declined, the protein was re-expressed from 4-6 h. Bcl-xL mRNA was up-regulated from 2 to 4 h in neonatal rat or mouse cardiac myocytes exposed to H2O2, consistent with the re-expression of protein. Four different untranslated first exons have been identified for the Bcl-x gene (exons 1, 1B, 1C, and 1D, where exon 1 is the most proximal and exon 1D the most distal to the coding region). All were detected in mouse or rat neonatal cardiac myocytes, but exon 1D was not expressed in adult mouse hearts. In neonatal mouse or rat cardiac myocytes, H2O2 induced the expression of exons 1B, 1C, and 1D, but not exon 1. These data demonstrate that the Bcl-x gene is selectively responsive to oxidative stress, and the response is mediated through distal promoter regions. PMID- 12721310 TI - What should be done about hepatitis-B-infected health-care workers? PMID- 12721311 TI - Immunoglobulins G could prevent adherence of Candida albicans to polystyrene and extracellular matrix components. AB - Immunocompromised patients are at high risk of developing Candida infections. Although cell-mediated immunity is generally believed to play the main role in defence against fungi, antibodies could also be effective in immune defence by different mechanisms of action. The adherence capacity of four strains of Candida albicans to polystyrene and to some extracellular matrix components was investigated after incubation of the yeasts with non-specific and specific anti C. albicans IgG. Experiments were carried out using a colorimetric method based upon the reduction of XTT tetrazolium (2,3-bis[2-methoxy-4-nitro-5-sulfophenyl] 2H-tetrazolium-5-carboxanilide) by mitochondrially active blastospores in the presence of menadione. Incubation of the yeasts with IgG, specific or not, caused a decrease in the capacity for adherence to the surfaces studied. There was no significant effect of the specificity of the tested antibodies on the reduction of adherence capacity. In conclusion, total IgG could play a role in blocking the binding of C. albicans to host and medical device surfaces. These results suggest that regular survey of levels of total IgG in patients suffering from severe hypogammaglobulinaemia could be of interest for the prevention of systemic candidiasis. PMID- 12721312 TI - Antibodies raised in animals against the Streptococcus agalactiae proteins c alpha and R4 and normal human serum antibodies target distinct epitopes. AB - The targets for normal human serum antibodies that react with proteins c(alpha) and R4 isolated from group B streptococci (GBS; Streptococcus agalactiae) have been studied and compared with the targets for murine monoclonal and rabbit polyclonal antibodies raised against these proteins. The proteins were extracted by trypsin digestion and purified by precipitations and gel filtration and testing was based on enzyme immunoassays. The immune antibodies showed specificity for the corresponding protein, targeted that protein in Western blotting and recognized their targets after heat treatment (100 degrees C) of the proteins. Human antibodies in a commercial gammaglobulin preparation targeted a site(s) common to c(alpha) and R4. This target failed to bind the antibodies in Western blotting and was destroyed by heating. c(alpha)- and R4-reactive antibodies in sera from healthy pregnant women recognized the common, heat-labile determinant(s), but contained little or no antibodies against the heat-stable c(alpha)- or R4-specific determinants. These results are consistent with the notions that (i) the normal human antibodies and the immunization-induced animal antibodies targeted different sites on the c(alpha) and R4 proteins and that (ii) the natural human antibodies targeted conformational epitopes and the immune antibodies targeted linear epitopes. These findings are important for further clarification of GBS immunology and immunoprotection in humans. PMID- 12721313 TI - Improvement of mupirocin E-test for susceptibility testing of Staphylococcus aureus. AB - Interpretation of the mupirocin E-test for low-level mupirocin-resistant Staphylococcus aureus strains has been improved by adding the indicator dye tetrazolium. E-tests were compared with agar dilution methods for assessing mupirocin susceptibility. MICs obtained by the agar dilution method and E-tests showed 89.3% agreement within 2 log(2) dilution criteria. The agreement between MICs increased to 100% in the 1 log(2) dilution definition when the indicator dye tetrazolium was added to the E-test. The use of the E-test with tetrazolium reduction is more accurate for determining mupirocin MICs for S. aureus strains. PMID- 12721314 TI - Heat-stable serogroup-specific proteins of Yersinia pseudotuberculosis. AB - A library of mAbs to the species- and serogroup-specific epitopes of Yersinia pseudotuberculosis serogroups I-VI was developed. These mAbs recognized linear sequential protein epitopes, as shown by ELISA and immunoblotting. Using the mAbs, Y. pseudotuberculosis was found to produce serogroup-specific proteins, whose synthesis was dependent on cultivation temperature. These proteins appeared to be parts of heat-stable O-antigens prepared by heating Y. pseudotuberculosis serogroups I-VI at 100 degrees C for 2 h, and are responsible for the protein serotype specificity of these bacteria. The high specificity of serogroup- or species-specific mAbs obtained in ELISA suggests that they may be effective for serotyping of Y. pseudotuberculosis strains or differentiation from other pathogenic yersiniae. PMID- 12721315 TI - Rapid quantification of hepatitis B virus DNA by real-time PCR using fluorescent hybridization probes. AB - A highly sensitive and rapid assay has been developed to quantify hepatitis B virus (HBV) DNA, based on the fluorescence resonance energy transfer principle and real-time PCR, using the LightCycler and a pair of specific fluorescent hybridization probes. This LightCycler real-time PCR assay (LC-PCR) detected HBV DNA in a linear range from 10(1) to 10(8) copies per reaction (250-2.5 x 10(9) copies ml(-1)), with a rapid PCR cycling time of 35 min. The assay was validated with two EUROHEP HBV DNA standards (ad and ay subtypes) and exhibited low intra assay (< 6 %) and inter-assay (< 16 %) variation for both subtypes over the complete range of 7 orders of magnitude. The assay was evaluated clinically using serum samples from 120 HBsAg(+) individuals and 45 healthy controls who were negative for both HBsAg and anti-HBc. Levels of HBV DNA were measured in these samples using both the LC-PCR and Digene Hybrid Capture II HBV DNA (HCII) assays. The prevalence rates for HBV DNA in the HBsAg(+) serum samples were respectively 95 % (114/120) and 56 % (67/120) by LC-PCR and HCII (P < 0.01). All 67 HCII positive samples tested positive with LC-PCR, while the 47 discordant samples showed low levels of HBV DNA (down to 265 copies ml(-1)), detectable only by the more sensitive LC-PCR assay. Levels of HBV DNA as measured by the two assays showed good correlation (r = 0.902; P < 0.001). The level of HBV DNA was significantly higher in HBeAg(+) than anti-HBe(+) samples (median 1.5 x 10(7) vs 4.6 x 10(4) copies ml(-1); P < 0.01). It is concluded that this LC-PCR assay is clinically useful for the rapid, sensitive and accurate measurement of HBV DNA. PMID- 12721317 TI - Aspirin inhibits Chlamydia pneumoniae-induced NF-kappa B activation, cyclo oxygenase-2 expression and prostaglandin E2 synthesis and attenuates chlamydial growth. AB - Infection with Chlamydia pneumoniae has been implicated as a potential risk factor for atherosclerosis. This study was designed to investigate the mechanisms of the anti-chlamydial activity of aspirin. A reporter gene assay for NF-kappa B activity, immunoblot analysis for cyclo-oxygenase (COX)-2 and radioimmunoassay for prostaglandin E(2) (PGE(2)) were performed. Following infection of HEp-2 cells with C. pneumoniae, NF-kappa B was activated, COX-2 was induced and PGE(2) was elevated. Aspirin inhibited NF-kappa B activation at a concentration of 0.1 mM, partially inhibited COX-2 induction and blocked PGE(2) synthesis completely. In addition, high doses of aspirin (1 and 2 mM) inhibited chlamydial growth in HEp-2 cells, decreasing the number and size of inclusion bodies; this effect could be overcome by adding tryptophan to the culture. Indomethacin also blocked the synthesis of PGE(2), but had no effect on COX-2 expression or chlamydial growth. These results indicate that aspirin not only has an anti-inflammatory activity through prevention of NF-kappa B activation but also has anti-chlamydial activity at high doses, possibly through depletion of tryptophan in HEp-2 cells. PMID- 12721316 TI - Expression stability of six housekeeping genes: A proposal for resistance gene quantification studies of Pseudomonas aeruginosa by real-time quantitative RT PCR. AB - Constantly expressed genes are used as internal controls in relative quantification studies. Suitable internal controls for such studies have not yet been defined for Pseudomonas aeruginosa. In this study, the genes ampC, fabD, proC, pbp-2, rpoD and rpoS of P. aeruginosa were compared in terms of expression stability by real-time quantitative RT-PCR. A total of 23 strains with diverse resistance phenotypes were studied. Stability of expression among the housekeeping genes was assessed on the basis of correlation coefficients, with the best-correlated pair accepted as being the most stable one. Eventually, proC and rpoD formed the most stable pair (r = 0.958; P < 0.001). Next, in four ciprofloxacin-selected nfxC-like mutants, levels of oprD, oprM and oprN mRNA were compared with those of their wild-type counterparts. The comparison was made after correcting the raw values by the geometric mean of the internal control genes proC and rpoD. The level of oprN mRNA was significantly up-regulated, while the oprD gene was down-regulated (although this difference was statistically insignificant), in the mutants. This expression pattern was consistent with that of the expected expression profile of nfxC-type mutants; this experiment therefore lends further support to the use of proC and rpoD genes simultaneously as internal controls for such studies. PMID- 12721318 TI - Inhibition of Helicobacter pylori growth in vitro by Bulgarian propolis: preliminary report. AB - Bee glue (propolis) possesses antimicrobial, anti-inflammatory, anaesthetic and immunostimulating activities. The aim of the study was to evaluate the inhibitory effect of Bulgarian propolis on Helicobacter pylori growth in vitro. Activity of 30% ethanolic extract of propolis (EEP) against 38 clinical isolates of H. pylori was evaluated by using the agar-well diffusion method. Ethanol was used as a control. In addition, the effect of propolis on the growth of 26 H. pylori and 18 Campylobacter strains was tested by the disc diffusion method. Mean diameters of H. pylori growth inhibition by the agar-well diffusion method, using 30, 60 or 90 microl EEP or 30 microl ethanol per well, were 17.8, 21.2, 28.2 and 8.5 mm, respectively. EEP was significantly more active than ethanol against H. pylori (P < 0.001). The results obtained by the disc diffusion method were similar. The use of moist propolis discs resulted in mean diameters of growth inhibition of 21.4 mm for H. pylori and 13.6 mm for Campylobacter spp. Dried propolis discs exhibited antibacterial effect against 73.1% of H. pylori isolates, with a considerable zone of growth inhibition (> or = 15 mm) in 36.4% of isolates. Using dried propolis discs resulted in mean diameters of growth inhibition of 12.4 mm for H. pylori and 11.6 mm for Campylobacter spp. In conclusion, Bulgarian propolis possesses considerable antibacterial activity against H. pylori, and can also inhibit the growth of Campylobacter jejuni and Campylobacter coli. The potential of propolis in the prevention or treatment of H. pylori infection is worth further extensive evaluation. PMID- 12721319 TI - Prevalence of extended-spectrum beta-lactamase-producing Gram-negative bacteria in septicaemic neonates in a tertiary care hospital. AB - The present study was undertaken to investigate the high incidence of multiresistant Gram-negative bacilli causing neonatal septicaemia. Samples of neonatal blood from 728 suspected cases were obtained in brain heart infusion broth with sodium polyanethol sulfonate. All Gram-negative rods isolated were subsequently subjected to routine antimicrobial susceptibility testing and tests for extended-spectrum beta-lactamase (ESBL) production, as per NCCLS recommendations. ESBL was detected in 86.6% of Klebsiella spp., 73.4% of Enterobacter spp. and 63.6% of Escherichia coli strains. It was also observed that 74.4-80.9% of these ESBL producers were resistant to cefotaxime and 47.6 59.5% were resistant to ceftazidime in routine susceptibility testing. Some ESBL producers (36.3-61.5%) were found to be susceptible to either or both cephalosporins used in this study. It is concluded that indiscriminate use of third-generation cephalosporins may be responsible for the selection of ESBL producing multiresistant strains in the neonatal intensive-care unit (NICU). PMID- 12721321 TI - Acute viral gastroenteritis: proportion and clinical relevance of multiple infections in Spanish children. AB - Dual infections associated with acute infectious diarrhoea and its microbiological, epidemiological and clinical findings have been evaluated in patients selected from a comprehensive survey of children under 4 years old, admitted to hospital emergency rooms from October 1996 to November 1997. A total of 820 children (433 males and 387 females) were enrolled. Stools were tested for rotavirus, adenovirus, astrovirus and bacterial enteropathogens. Patients were grouped according to age, and the seasonality of mixed infections was evaluated. Clinical trends and severity of gastrointestinal disease by Ruuska's score were also analysed. Mixed infections were identified in 39 cases (5 %), of which 23 were males and 16 were females. The majority of cases were in the 7-18-month age group (26 cases) and occurred in autumn (67 %). Virus-virus co-infections were more frequent (26/39) than virus-bacteria co-infections (13/39). More than two infectious agents were detected in only four cases. The most common viral co infections were rotavirus-astrovirus (13/26) and rotavirus-adenovirus (10/26). The present report is the first prospective analysis of clinical-epidemiological trends of dual infections in young Spanish children with acute viral gastroenteritis. Our results emphasize the clinical importance of mixed infections as a cause of severe diarrhoea in children. PMID- 12721322 TI - Misidentification of Brucella melitensis as Ochrobactrum anthropi by API 20NE. PMID- 12721320 TI - Molecular epidemiology of an outbreak of multiresistant Klebsiella pneumoniae in a Tunisian neonatal ward. AB - During the first quarter of 1996, a major outbreak of clinical infection caused by multiresistant Klebsiella pneumoniae (MRKP) occurred in the neonatal ward of the 'Maternite Wassila Bourguiba' in Tunis, Tunisia. In total, 32 isolates of MRKP, comprising 23 clinical isolates and nine surveillance isolates, were recovered during this period and analysed for epidemiological relatedness. The isolates were compared with 17 other isolates of MRKP that were recovered during 1995. Macrorestriction profiles of total genomic DNA following XbaI restriction endonuclease digestion were analysed by PFGE; this typing classified 56% of the 32 isolates recovered in 1996 into two major clusters. Cluster A included ten isolates from 1996 and three isolates recovered in 1995, whereas cluster B included eight isolates from the outbreak of 1996. The remaining isolates were genetically unrelated to those of clusters A and B; they constituted sporadic strains. The two major clusters were also evident using other molecular typing methods, such as random amplification of polymorphic DNA (RAPD) and enterobacterial repetitive intergenic consensus (ERIC)-PCR, where isolates of clusters A and B could be identified on the basis of their discriminative patterns. This investigation showed the predominance of two epidemic strains, and illustrated the ease with which MRKP strains can disseminate and persist within a single ward. PMID- 12721323 TI - The subtype-selective nicotinic acetylcholine receptor agonist SIB-1553A improves both attention and memory components of a spatial working memory task in chronic low dose 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated monkeys. AB - Monkeys that receive chronic low dose (CLD) 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) administration develop deficits in spatial delayed response task performance. The present study examined the extent to which SIB 1553A [(+/-)-4-[[2-(1-methyl-2-pyrrolidinyl)ethyl]thio]phenol hydrochloride], a novel neuronal nicotinic acetylcholine receptor (nAChR) agonist with selectivity for beta4 subunit-containing nAChRs, could counteract this cognitive deficit produced by CLD MPTP exposure. Prior to MPTP treatment, monkeys displayed a delay dependent decrement in performance on a variable delayed response task. CLD MPTP treatment caused a shift to a delay-independent pattern of responding on this task, such that short-delay trials were performed as poorly as long-delay trials. At lower doses (e.g., 0.025 mg/kg), SIB-1553A significantly improved performance on short-delay trials but only at 24 h after drug administration. At higher doses (e.g., 0.50 mg/kg), SIB-1553A significantly improved performance on both short- and long-delay trials at both 20 min and 24 h after drug administration. When tested 24 h after drug administration, monkeys performed long-delay trials with greater accuracy than they did under normal (pre-MPTP) conditions. These results suggest that at lower doses, SIB-1553A may be more effective in improving attentional deficits associated with CLD MPTP exposure, whereas at higher doses, SIB-1553A may effectively improve both attentional and memory performance. PMID- 12721324 TI - Involvement of endocytic organelles in the subcellular trafficking and localization of riboflavin. AB - Previous studies by our laboratory have suggested the potential role of receptor mediated endocytosis components in the cellular translocation of riboflavin (vitamin B2). To delineate the intracellular compartments and events involved in the internalization of riboflavin, we synthesized a rhodamine-labeled riboflavin conjugate to monitor its movement via fluorescent microscopy. Cellular uptake studies in BeWo cells show that rhodamine-riboflavin conjugate exhibits similar ligand affinity toward the putative riboflavin transport system as [3H]riboflavin, whereas rhodamine does not significantly interfere with its internalization mechanism. Microscope analysis reveals rapid internalization of the rhodamine-riboflavin conjugate via a riboflavin-specific process into acidic vesicular compartments throughout the cells. The intracellular punctate distribution is comparable with that of fluorescein isothiocyanate (FITC) transferrin, a well characterized receptor-mediated endocytosis substrate. Double labeling fluorescence microscopy studies further confirm that with 10 min of internalization, rhodamine-riboflavin conjugate substantially concentrates within vesicular structures associated with clathrin, rab5, FITC-transferrin, and the acidotropic marker LysoTracker Blue. In summary, our studies provide, for the first time, direct morphological evidence of the involvement of endocytosis machinery in the intracellular trafficking of riboflavin. The subcellular localization of rhodamine-riboflavin conjugate suggests that, under the experimental conditions in this study, the internalization of riboflavin follows a classical receptor-mediated endocytosis pathway. PMID- 12721325 TI - Renal action of acute chloroquine and paracetamol administration in the anesthetized, fluid-balanced rat. AB - Chloroquine induces diuresis, natriuresis, and an increase in glomerular filtration rate (GFR) in the rat. These responses are modified in rats with analgesic nephropathy induced by long-term paracetamol (acetaminophen) administration. Here, the effects of acute paracetamol treatment on renal function and the response to chloroquine are reported. Under intraval anesthesia (100 mg kg-1) male Sprague-Dawley rats (n = 6/group) were infused with 2.5% dextrose for 3 h. After a control hour, they received either vehicle, chloroquine (0.04 mg h-1), paracetamol (priming dose of 210 mg kg-1 followed by 110 mg kg-1h 1) or chloroquine and paracetamol over the next hour. Compared with vehicle, chloroquine infusion resulted in increases in GFR (2.4 +/- 0.3 versus 4.8 +/- 0.6 ml min-1), urine flow (4.2 +/- 0.3 versus 10.4 +/- 0.7 ml h-1), and sodium excretion (47.7 +/- 4.1 versus 171.2 +/- 18.6 micromol h-1) and a reduction in urine osmolality (223.2 +/- 5.9 versus 121.7 +/- 23.9 mOsM kg-1). Paracetamol reduced sodium excretion but had no effect on urine flow, GFR, or urine osmolality. When combined, paracetamol blocked the chloroquine-induced diuresis (3.9 +/- 0.7 ml h-1) and natriuresis (22.6 +/- 8.5 micromol h-1), attenuated the increase in glomerular filtration rate (3.5 +/- 0.2 ml min-1), and raised urine osmolality (280.0 +/- 22.8 mOsM kg-1). The differing effects of acute and long term paracetamol treatment on basal and chloroquine-mediated renal function suggest that the length of prior exposure to paracetamol, and thus the presence of analgesic nephropathy, is an important determinant of the renal response to chloroquine. PMID- 12721326 TI - Ethanol excitation of dopaminergic ventral tegmental area neurons is blocked by quinidine. AB - The dopaminergic (DA) neurons in the ventral tegmental area (VTA) are important for the reinforcing effects of ethanol. We have shown that ethanol directly excites DA VTA neurons and reduces the afterhyperpolarization (AHP) that follows spontaneous action potentials in these neurons. These data suggested that ethanol may be increasing the firing rate of DA VTA neurons by modulating currents that contribute to the AHP, either by reducing a K+ current or by increasing the inward current Ih. In the present study, different blockers of K+ channels and Ih were tested to determine whether any could prevent the ethanol excitation of DA VTA neurons. Extracellular single-unit recordings and whole-cell patch-clamp recordings were made from DA VTA neurons in brain slices from Fischer-344 rats and ethanol (40-120 mM) and channel blockers were applied in the bath. Ethanol excitation was not reduced by blockade of Ih with cesium (5 mM) or ZD7288 (30 microM), or by block of G-protein-coupled inwardly rectifying K+ channels with barium (500 microM). Tetraethylammonium (TEA) ion (2-10 mM), which blocks the large conductance calcium-dependent potassium K+ current and some types of delayed rectifier currents, had no effect on the ethanol-induced excitation. Interestingly, ethanol excitation of DA VTA neurons was blocked by quinidine (20 80 microM), a drug that blocks many types of delayed rectifier K+ channels, including some insensitive to TEA. This effect of quinidine was concentration dependent and reversible. These results suggest that ethanol excites DA VTA neurons by reducing a quinidine-sensitive K+ current. PMID- 12721327 TI - Expression and molecular pharmacology of the mouse CRTH2 receptor. AB - Prostaglandin D2 (PGD2), the predominant prostanoid produced by activated mast cells, is implicated in a variety of allergic diseases. PGD2 exerts its effects through two G-protein coupled receptors, DP and CRTH2. PGD2 mediates chemotaxis of eosinophils, basophils, and Th2 cells via CRTH2-evoked signaling, suggesting a role for this receptor in allergic disease. To characterize the mouse CRTH2 ortholog (mCRTH2), we amplified the mCRTH2 receptor gene and expressed it in HEK293 cells. Saturation ligand binding isotherms demonstrated high-affinity binding of [3H]PGD2, with a Kd of 8.8 +/- 0.8 nM. Competition binding assays with a panel unlabeled prostanoids demonstrated an order of affinity of 13,14-dihydro 15-keto-PGD2 (DK-PGD2) >or= 15-deoxy-Delta12,14-PGJ2 (15d-PGJ2) >or= PGD2 >or= PGJ2. [3H]PGD2 binding was also displaced by the nonsteroidal anti-inflammatory drug indomethacin, with a Ki value of 1.04 +/- 0.13 microM. No [3H]PGD2 displacement was detected using fluribrofen, ibuprofen, or aspirin as competitors at concentrations of up to 30 microM. PGD2, DK-PGD2, 15d-PGJ2, and indomethacin each inhibited intracellular cAMP generation in stable transfectant ER293/mCRTH2 cells through a pertussis toxin (PTX) sensitive pathway, consistent with mCRTH2 coupling to a Gi heterotrimeric G-protein. Activation of mCRTH2 elicited chemotaxis of ER293/mCRTH2 cells in response to PGD2, indomethacin, and 15d-PGJ2. mCRTH2-dependent chemotaxis was inhibited by PTX and wortmannin, indicating dependence on Gi and PI 3-kinase signal transduction pathways. These data provide the first pharmacological and functional characterization of the mouse CRTH2 receptor. PMID- 12721329 TI - Threonine-205 in the F helix of p450 2B1 contributes to androgen 16 beta hydroxylation activity and mechanism-based inactivation. AB - Four mutants of Thr-205 in cytochrome p450 2B1 were constructed and expressed in Escherichia coli. The Ser-, Ala-, and Val-mutants displayed stable reduced CO difference spectra and were able to metabolize 7-ethoxy-4 (trifluoromethyl)coumarin, testosterone, androstenedione, and benzphetamine. The Arg-mutant displayed an unstable reduced CO difference spectrum at 450 nm, was concomitantly converted to a denatured form with a peak at 422 nm, and showed no catalytic activity with any of the four substrates tested. The Ser-mutant displayed activity and metabolite profiles for testosterone and androstenedione similar to those of the wild-type p450 2B1 (WT). Substitution of Thr-205 with Ala or Val markedly suppressed the 16 beta-hydroxylation activity but exhibited little effect on the 16 alpha-hydroxylation activity for testosterone and androstenedione. Because 16 beta-hydroxylation activity of androgens is a specific p450 2B subfamily marker and residue 205 is located in the F helix, which forms the ceiling of the active site, we postulate that the gamma-hydroxyl side chain of Thr may play an important role in directing the 16 beta-face of testosterone and androstenedione toward the active site. Surprisingly, the Val mutant retained full activity for benzphetamine demethylation. When mechanism based inactivators for p450 2B1 were used to evaluate the susceptibility to inactivation, the Val-mutant was resistant to inactivation by 17 alpha ethynylestradiol and less sensitive to inactivation by 2-ethynylnaphthalene compared with the WT enzyme. Our results demonstrate the importance of Thr-205 in determining substrate specificity and product formation as well as in influencing the susceptibility of p450 2B1 to mechanism-based inactivators. PMID- 12721330 TI - Functional melatonin receptors in rat ovaries at various stages of the estrous cycle. AB - This study investigated the receptor mechanism(s) by which the hormone melatonin directly affects ovarian function. Expression of MT1 and MT2 melatonin receptor mRNA was detected in the rat ovaries both by reverse transcriptase-polymerase chain reaction and in situ hybridization with digoxigenin-labeled oligoprobes. Specific 2-[125I]iodomelatonin binding was significantly higher in ovarian tissue from animals sacrificed during proestrus than in metestrus, suggesting regulation of melatonin receptors by estrogens. Additionally, basal and melatonin-mediated stimulation of guanosine 5'-O-(3-[35S]thio)triphosphate ([35S]GTPgammaS) binding to ovarian sections was higher in proestrus compared with metestrus. During proestrus, both luzindole (0.1 microM) and 4-phenyl-2-propionamidotetraline (4P PDOT) (0.1 microM), acting as inverse agonists, inhibited basal [35S]GTPgammaS binding to ovarian sections, suggesting the presence of MT1 constitutively active melatonin receptors. In primary cultures of ovarian granulosa cells, melatonin inhibited forskolin-stimulated cAMP accumulation through activation of Gi-coupled melatonin receptors. This inhibition was blocked by both, luzindole, and 4P-PDOT, acting as competitive receptor antagonists. Exposure of granulosa cells in culture to 17beta-estradiol seems to alter the state of melatonin receptor coupling. Indeed, the efficacy of 4P-PDOT on forskolin-stimulated cAMP formation was reversed from an MT2 partial agonist in vehicle-treated cells to that of an MT1 inverse agonist in 17beta-estradiol (0.1 microM)-treated granulosa cells. We conclude that MT1 and MT2 melatonin receptors expressed in antral follicles and corpus luteum may affect steroidogenesis through cAMP-mediated signaling. These results underscore the implications of the levels of ovarian estrogen when melatonin receptor ligands are used as therapeutic agents. PMID- 12721328 TI - Chronic fluoxetine differentially affects 5-hydroxytryptamine (2A) receptor signaling in frontal cortex, oxytocin- and corticotropin-releasing factor containing neurons in rat paraventricular nucleus. AB - Differential adaptive changes in serotonin2A [5-hydroxytryptamine (5-HT)2A] receptor signaling during treatment may be one mechanism involved in the latency of therapeutic improvement with antidepressants, such as fluoxetine. We examined the effects of fluoxetine (2, 3, 7, 21, or 42 days) on hypothalamic 5-HT2A receptor signaling. The hormone responses to an injection of the 5-HT2A receptor agonist (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-amino-propane HCl (DOI) were used as an index of hypothalamic 5-HT2A receptor function. Treatment with fluoxetine for 21 or 42 days produced diminished adrenocorticotropic hormone (ACTH) and oxytocin (but not corticosterone) responses to DOI injections (2.5 mg/kg i.p.; 15 min postinjection). Regulators of G protein signaling 4 and Galphaq protein levels in the hypothalamic paraventricular nucleus were not altered during fluoxetine treatment. Because previous studies indicate that treatment with fluoxetine for 21 days resulted in increased hormone responses to DOI when measured at 30 min after injection, we examined the effect of fluoxetine (21 days) on DOI-induced increase hormone levels at 15, 30, and 60 min after DOI injection. Fluoxetine decreased the oxytocin response at 15 but not at 30 min post-DOI injection, and potentiated the ACTH and corticosterone responses at 30 min post-DOI injection. For comparison, we examined the effect of fluoxetine on 5 HT2A receptor-mediated increase in phospholipase C (PLC) activity in the frontal cortex. 5-HT-stimulated, but not guanosine 5'-O-(3-thio)triphosphate-stimulated PLC activity was increased after 21 days of fluoxetine-treatment. Overall, these results indicate that chronic fluoxetine treatment can potentiate 5-HT2A receptor signaling in frontal cortex but differentially alters 5-HT2A receptor signaling in oxytocin-containing neurons and corticotropin-releasing factor-containing neurons in the paraventricular nucleus. PMID- 12721331 TI - Role of nitric-oxide synthase isoforms in nitrous oxide antinociception in mice. AB - Exposure of mice to the anesthetic gas N2O evokes a prominent antinociceptive effect that is sensitive to antagonism by nonselective nitric-oxide synthase (NOS) inhibitors. The present study was conducted to identify whether a specific NOS isoform is implicated in N2O antinociception in mice. In the abdominal constriction test, exposure of mice to 25, 50, and 70% N2O resulted in a concentration-dependent antinociceptive effect that persisted for up to 6 min following removal of the mice from the N2O atmosphere into room air. This N2O antinociceptive effect was antagonized by pretreatment with S-methyl-l thiocitrulline (SMTC) and higher doses of l-N5-(1-iminoethyl)-ornithine (l-NIO), which reportedly inhibit the neuronal and endothelial isoforms of NOS, respectively. Nevertheless, the N2O-induced antinociception was unaffected by pretreatment with low doses of either SMTC or l-NIO or by pretreatment with 2 amino-5,6-dihydro-6-methyl-4H-1,3-thiazine (AMT), which selectively inhibits inducible NOS. The s.c. pretreatment with SMTC and l-NIO reduced brain NOS activity in a dose-dependent manner, whereas AMT had no such effect. Moreover, in blood pressure experiments, SMTC increased SBP in dose-unrelated fashion, whereas l-NIO showed an appreciably weaker but dose-related increase in SBP. The i.c.v. pretreatment with SMTC also reduced N2O antinociception and brain NOS activity without increasing of SBP. These results suggest that it is the neuronal isoform of NOS that is involved in mediation of the antinociceptive effect of N2O in the mice. PMID- 12721332 TI - Modulation of P-glycoprotein transport activity in the mouse blood-brain barrier by rifampin. AB - The objective of the present study was to examine the time course and concentration dependence of modulation of P-glycoprotein (P-gp) activity in the blood-brain barrier (BBB) with consequent influence on substrate uptake into brain tissue. Potential P-gp inducers (rifampin and morphine) were administered subchorionically to P-gp-competent [mdr1a(+/+)] mice to induce P-gp expression in brain; the impact of rifampin pretreatment on brain penetration of verapamil also was evaluated with an in situ brain perfusion technique. In addition, the effect of single-dose rifampin on P-gp BBB transport activity was assessed with brain perfusion using verapamil and quinidine as model P-gp substrates. Chronic exposure to rifampin or morphine induced P-gp expression in mouse brain to a modest extent. However, single-dose rifampin treatment increased the brain uptake of verapamil and quinidine in mdr1a(+/+) mice in a dose- and concentration dependent manner, consistent with P-gp inhibition. Maximum inhibition of P-gp mediated efflux of verapamil by rifampin pretreatment in vivo (150 mg/kg) was approximately 55%, whereas there was only approximately 12% inhibition of P-gp mediated efflux of quinidine at that rifampin dose. Coperfusion of rifampin at a concentration of 500 microM abolished P-gp-mediated efflux of verapamil at the BBB. However, only approximately 40% inhibition of P-gp-mediated efflux of quinidine was observed with coperfusion of rifampin, even at a 2-fold higher rifampin concentration (1000 microM). The present studies demonstrate that P-gp function at the BBB can be modulated by rifampin in a dose- and concentration dependent manner. The degree to which rifampin inhibits P-gp-mediated transport is dependent on the substrate molecule. PMID- 12721333 TI - Buprenorphine blocks epsilon- and micro-opioid receptor-mediated antinociception in the mouse. AB - Antagonistic properties of buprenorphine for epsilon- and micro -opioid receptors were characterized in beta-endorphin- and [d-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin (DAMGO)-induced antinociception, respectively, with the tail-flick test in male ICR mice. epsilon-Opioid receptor agonist beta-endorphin (0.1-1 micro g), micro opioid receptor agonist DAMGO (0.5-20 ng), or buprenorphine (0.1-20 micro g) administered i.c.v. dose dependently produced antinociception. The antinociception induced by 10 micro g of buprenorphine given i.c.v. was completely blocked by the pretreatment with beta-funaltrexamine (beta-FNA) (0.3 micro g i.c.v.), indicating that the buprenophine-induced antinociception is mediated by the stimulation of the micro -opioid receptor. The antinociceptive effects induced by beta-endorphin (1 micro g i.c.v.) and DAMGO (16 ng i.c.v.) were dose dependently blocked by pretreatment with smaller doses of buprenorphine (0.001-1 micro g i.c.v.), but not by a higher dose of buprenorphine (10 micro g i.c.v.). beta-FNA at a dose (0.3 micro g i.c.v.) that strongly attenuated DAMGO induced antinociception had no effect on the antinociception produced by beta endorphin (1 micro g i.c.v.). However, pretreatment with buprenorphine (0.1-10 micro g) in mice pretreated with this same dose of beta-FNA was effective in blocking beta-endorphin-induced antinociception. beta-FNA was 226-fold more effective at antagonizing the antinociception induced by DAMGO (16 ng i.c.v.) than by beta-endorphin (1 micro g i.c.v.). The antinociception induced by delta opioid receptor agonist [d-Ala2]deltorphin II (10 micro g i.c.v.) or kappa1 opioid receptor agonist trans-3,4-dichloro-N-methyl-N-(2-[1 pyrrolidinyl]cyclohexyl)benzeneacetamine methanesulfonate salt [(-)-U50,488H] (75 micro g i.c.v.) was not affected by pretreatment with buprenorphine (0.1-1.0 micro g i.c.v.). It is concluded that buprenorphine, at small doses, blocks epsilon-opioid receptor-mediated beta-endorphin-induced antinociception and micro -opioid receptor-mediated DAMGO-induced antinociception, and at high doses produces a micro -opioid receptor-mediated antinociception. PMID- 12721334 TI - Nociceptin/orphanin FQ and its receptor--potential targets for pain therapy? AB - The neuropeptide nociceptin, also called orphanin FQ (N/OFQ), is the endogenous agonist of the N/OFQ peptide receptor (NOP receptor). Both N/OFQ and the NOP receptor share a high degree of homology with classical opioid peptides and opioid receptors, respectively, and use similar signal transduction pathways as classical opioids. The NOP receptor has thus been regarded as the fourth member of the opioid receptor family. Despite this close relationship, 7 years of research have demonstrated that the N/OFQ system has a distinct pharmacological profile and serves different physiological functions. In particular, its role in the control of pain and analgesia at different levels of integration appears quite different from that of classical opioids. The recent development of specific antagonists at the NOP receptor and of NOP receptor or N/OFQ precursor knock-out mice have generated new insights into the role of N/OFQ in pain processing and help to evaluate the N/OFQ-NOP system as a potential target for new analgesic drugs. PMID- 12721335 TI - BM-573, a dual thromboxane synthase inhibitor and thromboxane receptor antagonist, prevents pig myocardial infarction induced by coronary thrombosis. AB - The aim of this study was to characterize the effects of BM-573 [N-terbutyl-N'-[2 (4'-methylphenylamino)-5-nitro-benzenesulfonyl] urea], a novel dual thromboxane A2 receptor antagonist and thromboxane synthase inhibitor, on myocardial infarction induced by topical ferric chloride (FeCl3) application to the left anterior descending (LAD) coronary artery in anesthetized pigs. All control animals (n = 6) developed an occlusive thrombus in the LAD coronary artery. The mean infarct size, revealed by triphenyl tetrazolium chloride (TTC), and the area at risk, evidenced by Evans blue, corresponded to 35.3 +/- 2.2 and 36.9 +/- 2.1% of the left ventricular mass, respectively. In the BM-573-treated group (n = 6), a drug infusion (10 mg. kg-1. h-1) started 30 min before FeCl3 application and continued throughout the experimentation. Among the BM-573-treated group, four pigs did not develop coronary artery thrombus and their myocardium appeared healthy. Histopathological examination of FeCl3-injured coronary artery revealed an occlusive and adherent thrombus in control group, while pretreatment with BM 573 prevented thrombus formation. In infarcted zones, lack of desmin staining and muscle structure disorganization were obvious. Depletion of myocardial ATP content was observed in the myocardial necrotic region of the control group, but not in myocardial samples of BM-573-treated pigs that did not develop myocardial infarction. When BM-573 prevented LAD artery occlusion, the area under the curve of plasmatic troponin T was reduced by 77% over 6 h. These data suggest that BM 573 could be useful for the prevention of myocardial infarction. PMID- 12721336 TI - N-(4-Tertiarybutylphenyl)-4-(3-cholorphyridin-2-yl)tetrahydropyrazine -1(2H) carbox-amide (BCTC), a novel, orally effective vanilloid receptor 1 antagonist with analgesic properties: II. in vivo characterization in rat models of inflammatory and neuropathic pain. AB - The vanilloid receptor 1 (VR1) is a cation channel expressed predominantly by nociceptive sensory neurons and is activated by a wide array of pain-producing stimuli, including capsaicin, noxious heat, and low pH. Although the behavioral effects of injected capsaicin and the VR1 antagonist capsazepine have indicated a potential role for VR1 in the generation and maintenance of persistent pain states, species differences in the molecular pharmacology of VR1 and a limited number of selective ligands have made VR1 difficult to study in vivo. N-(4 Tertiarybutylphenyl)-4-(3-cholorphyridin-2-yl)tetrahydropryazine-1(2H)-carbox amide (BCTC) is a recently described inhibitor of capsaicin- and acid-mediated currents at rat VR1. Here, we report the effects of BCTC on acute, inflammatory, and neuropathic pain in rats. Administration of BCTC (30 mg/kg p.o.) significantly reduced both mechanical and thermal hyperalgesia induced by intraplantar injection of 30 micro g of capsaicin. In rats with Freund's complete adjuvantinduced inflammation, BCTC significantly reduced the accompanying thermal and mechanical hyperalgesia (3 mg/kg and 10 mg/kg p.o., respectively). BCTC also reduced mechanical hyperalgesia and tactile allodynia 2 weeks after partial sciatic nerve injury (10 and 30 mg/kg p.o.). BCTC did not affect motor performance on the rotarod after administration of doses up to 50 mg/kg p.o. These data suggest a role for VR1 in persistent and chronic pain arising from inflammation or nerve injury. PMID- 12721337 TI - Hyperlocomotive and discriminative stimulus effects of cocaine are under the control of serotonin(2C) (5-HT(2C)) receptors in rat prefrontal cortex. AB - The serotonin2C (5-hydroxytryptamine2C; 5-HT2C) receptor (5-HT2CR) is found in abundance in dopamine (DA) mesocorticolimbic pathways and is one of the important target proteins that modulates the behavioral effects of cocaine. In the present study, the hypothesis was tested that the 5-HT2CR in the prefrontal cortex (PFC) may control either spontaneous or cocaine-evoked locomotor activity as well as the discriminative stimulus properties of cocaine. In male Sprague-Dawley rats implanted with bilateral cannulae aimed at the PFC, local microinjections of the preferential 5-HT2CR agonist 6-chloro-2-(1-piperazinyl)pyrazine hydrochloride (MK 212) (0.05-0.5 microg/side) did not alter spontaneous activity, but dose dependently decreased horizontal hyperactivity evoked by cocaine (10 mg/kg i.p.). Given alone, the selective 5-HT2CR antagonist 8-[5-(2,4-dimethoxy-5-(4 trifluorophenylsulfonamido)phenyl-5-oxopentyl]-1,3,8-triazo-spiro[4.5]decane-2,4 dione hydrochloride (RS 102221) (5 microg/side) increased basal locomotor activity of rats expressed in the vertical plane. Microinjections of RS 102221 (5 microg/side, but not 0.15-1.5 microg/side) significantly enhanced the horizontal activity induced by cocaine (10 mg/kg). In rats trained to discriminate cocaine (10 mg/kg i.p.) from saline (i.p.) in a two-lever, water-reinforced fixed ratio 20 task, intra-PFC microinjections of MK 212 (0.05 and 0.5 microg/side) did not substitute for cocaine, but attenuated the stimulus effects of cocaine. However, intra-PFC microinjections of RS 102221 (1.5 and 5 microg/side) evoked 13 and 40% cocaine-lever responding when tested alone and enhanced the recognition of cocaine. These data indicate that the PFC is a brain site at which the 5-HT2CR exerts an inhibitory control over the hyperactive and discriminative stimulus effects of cocaine known to be dependent upon activation of the DA mesoaccumbens circuit. PMID- 12721338 TI - N-(4-tertiarybutylphenyl)-4-(3-chloropyridin-2-yl)tetrahydropyrazine -1(2H) carbox-amide (BCTC), a novel, orally effective vanilloid receptor 1 antagonist with analgesic properties: I. in vitro characterization and pharmacokinetic properties. AB - Vanilloids such as capsaicin have algesic properties and seem to mediate their effects via activation of the vanilloid receptor 1 (VR1), a ligand-gated ion channel highly expressed on primary nociceptors. Although blockade of capsaicin induced VR1 activation has been demonstrated in vitro and in vivo with the antagonist capsazepine, efficacy in rat models of chronic pain has not been observed with this compound. Here, we describe the in vitro pharmacology of a highly potent VR1 antagonist, N-(4-tertiarybutylphenyl)-4-(3-chloropyridin-2 yl)tetrahydropyrazine-1(2H)-carbox-amide (BCTC). Similar to capsazepine, this compound inhibits capsaicin-induced activation of rat VR1 with an IC50 value of 35 nM. Interestingly however, BCTC also potently inhibits acid-induced activation of rat VR1 (IC50 value of 6.0 nM), whereas capsazepine is inactive. Similarly, in the rat skin-nerve preparation both BCTC and capsazepine block capsaicin-induced activation, whereas the response to acidification is inhibited by BCTC, but not by capsazepine. Specificity for VR1 was demonstrated against 63 other receptor, enzyme, transporter, and ion channel targets. BCTC was orally bioavailable in the rat, demonstrating a plasma half-life of approximately 1 h and significant penetration into the central nervous system. Thus, BCTC is a high potency, selective VR1 antagonist that, unlike capsazepine, has potent blocking effects on low pH-induced activation of rat VR1. These properties make it a more suitable candidate than capsazepine for testing the role played by VR1 in rat models of human disease. PMID- 12721339 TI - Integration of science into orthopaedic practice: implications for solving the problem of articular cartilage repair. PMID- 12721340 TI - Outcome of untreated traumatic articular cartilage defects of the knee: a natural history study. AB - BACKGROUND: Articular cartilage damage has been reported in 23% of knees with an acute anterior cruciate ligament (ACL) injury and 54% of those with chronic ACL laxity. Because the purpose of surgery is to reconstruct the ACL, the chondral lesion is usually an incidental finding. It is not known if any of the numerous treatments that have been recommended for chondral defects alters the natural history of the untreated lesion. We sought to determine what effect, if any, an isolated articular cartilage defect observed at the time of ACL reconstruction would have on the radiographic, subjective, and objective results after surgery if no intervention was performed on the cartilage lesion itself. METHODS: From 1987 to 1999, 2770 ACL reconstructions were performed, and 125 of them were done in patients who had an articular cartilage defect of Outerbridge grade 3 or 4 but had both menisci intact. The mean defect size was 1.7 cm (2) (range, 0.5 to 6.5 cm (2) ). Postoperative rehabilitation was not altered because of the chondral defect, and patients were allowed full weight-bearing and the full range of motion of which they were capable. A control group of patients matched on the basis of sex and age at surgery was identified from the database. No patient in the control group had a chondral defect or meniscal tear. Patients were evaluated at one, two, and five years after surgery and every five years thereafter with use of the IKDC (International Knee Documentation Committee) criteria, modified Noyes subjective questionnaire, and radiographs. RESULTS: Subjective follow-up was carried out more than two years after surgery (mean time, 8.7 years after surgery) for 101 patients. The results of objective evaluation were available for fifty-two patients, at a mean of 6.3 years. The patients in the control group had significantly higher subjective scores than did the patients with a defect in the medial compartment (mean, 95.2 points versus 94.0 points; p = 0.0451) and those with a defect in the lateral compartment (mean, 95.9 points versus 92.8 points; p = 0.0047). There was no significant correlation between larger defect size and lower subjective scores (p = 0.2543). The distribution of IKDC radiographic ratings was not significantly different between the groups. At least 79% of the patients in both groups returned to jumping, twisting, and pivoting sports at least at the recreational level. CONCLUSIONS: While statistical analysis revealed a difference in subjective scores between the defect and control groups, an average of 93 points for the patients with a lateral defect and 94 points for those with a medial defect indicates that most patients have very few symptoms. This study provides a baseline of information that can be used to compare the results of procedures designed to treat articular cartilage defects. PMID- 12721341 TI - Treatment of osteochondritis dissecans of the knee with autologous chondrocyte transplantation: results at two to ten years. AB - BACKGROUND: Osteochondritis dissecans of the knee is a challenging clinical problem. We previously reported on the early successful results of autologous chondrocyte transplantation for the treatment of focal cartilage defects. The purpose of the present study was to assess the intermediate to long-term results of this technique in a large group of patients with osteochondritis dissecans. METHODS: Fifty-eight patients with radiographically documented osteochondritis dissecans of the knee underwent treatment with autologous chondrocyte transplantation between 1987 and 2000 and were assessed clinically with use of standard rating scales. Twenty-two patients consented to arthroscopic second-look evaluation of graft integrity. RESULTS: The mean age of the patients at the time of autologous chondrocyte transplantation was 26.4 years (range, fourteen to fifty-two years). Seven patients were less than eighteen years old. Thirty-five patients (60%) had juvenile-onset disease, and forty-eight patients (83%) had had a mean of 2.1 prior operations. The defect was located on the medial femoral condyle in thirty-nine patients and on the lateral femoral condyle in nineteen. The mean lesion size was 5.7 cm (2) (range, 1.5 to 12.0 cm (2) ), and the mean defect depth was 7.8 mm (range, 4 to 15 mm). After a mean duration of follow-up of 5.6 years, 91% of the patients had a good or excellent overall rating on the basis of a clinician evaluation and 93% had improvement on a patient self assessment questionnaire. The Tegner-Wallgren, Lysholm, and Brittberg-Peterson VAS scores were all improved. The macroscopic quality of graft integrity averaged 11.2 on a 12-point scale, with only one graft having a score of <9 points. Two patients had a failure of treatment in the early postoperative period. Only one patient who had had a good or excellent rating at two years had a decline in clinical status at the time of the latest follow-up. CONCLUSIONS: Treatment of osteochondritis dissecans lesions of the knee with autologous chondrocyte transplantation produces an integrated repair tissue with a successful clinical result in >90% of patients. We recommend the wider use of autologous chondrocyte transplantation for this condition. PMID- 12721342 TI - Autologous osteochondral mosaicplasty for the treatment of full-thickness defects of weight-bearing joints: ten years of experimental and clinical experience. AB - BACKGROUND: The successful treatment of chondral and osteochondral defects of the weight-bearing surfaces is a challenge for orthopaedic surgeons. Autologous osteochondral transplantation is one method that can be used to create hyaline or hyaline-like repair in the defect area. This paper describes the results after ten years of clinical experience with autologous osteochondral mosaicplasty. METHODS: Clinical scores, imaging techniques, arthroscopy, histological examination of biopsy samples, and cartilage stiffness measurements were used to evaluate the clinical outcomes and quality of the transplanted cartilage in 831 patients undergoing mosaicplasty. RESULTS: According to these investigations, good-to-excellent results were achieved in 92% of the patients treated with femoral condylar implantations, 87% of those treated with tibial resurfacing, 79% of those treated with patellar and/or trochlear mosaicplasties, and 94% of those treated with talar procedures. Long-term donor-site disturbances, assessed with use of the Bandi score, showed that patients had 3% morbidity after mosaicplasty. Sixty-nine of eighty-three patients who were followed arthroscopically showed congruent gliding surfaces, histological evidence of the survival of the transplanted hyaline cartilage, and fibrocartilage filling of the donor sites. Complications of the surgery included four deep infections and thirty-six painful postoperative hemarthroses. CONCLUSIONS: On the basis of these promising results and those of other similar studies, autologous osteochondral mosaicplasty appears to be an alternative for the treatment of small and medium-sized focal chondral and osteochondral defects of the weight-bearing surfaces of the knee and other weight-bearing synovial joints. PMID- 12721343 TI - Long-term follow-up of fresh tibial osteochondral allografts for failed tibial plateau fractures. AB - BACKGROUND: The management of large posttraumatic full-thickness osteochondral defects in the proximal part of the tibia remains a challenge. The goal of treatment is a pain-free range of motion of the knee that provides enduring function and enables a young patient to participate in a wide range of activities. The use of fresh osteochondral allograft transplantation for tibial plateau lesions has been well documented. The purpose of the present study was to assess the survivorship and the long-term functional outcome following fresh osteochondral transplantation for unipolar posttraumatic tibial plateau defects in young, high-demand patients. METHODS: A cohort study was carried out to assess outcome in patients who had undergone fresh tibial osteochondral grafting for the treatment of a posttraumatic defect that measured at least 3 cm in diameter and 1 cm in depth. Sixty-five (97%) of sixty-seven patients were identified and were evaluated clinically and radiographically. A modified Hospital for Special Surgery (HSS) score was calculated for each patient. Radiographic examination was directed toward the identification of graft union, resorption, or collapse. Degeneration of the joint and alignment of the limb (on standing radiographs) was assessed. The end points that defined survivorship were an HSS score of <70 points, a patient's decision to undergo knee arthroplasty, or revision of the graft for any reason. RESULTS: At a mean of twelve years, forty-four patients had an intact graft and twenty-one had had conversion to a total knee arthroplasty. The mean modified HSS Score for the patients with an intact graft was 85 points. Radiographs, reviewed for thirty-five of the forty-four patients with an intact graft, revealed union of the graft to host bone in all cases and an 8.6% prevalence of graft collapse in excess of 3 mm. Forty percent of these thirty five patients demonstrated moderate to severe degenerative changes. Kaplan-Meier survivorship analysis showed that the survival rate was 95% at five years, 80% at ten years, 65% at fifteen years, and 46% at twenty years. A trend toward increased survivorship (p = 0.08) was seen among patients who had undergone meniscal transplantation in conjunction with osteochondral grafting. CONCLUSIONS: Fresh osteochondral allografts for large traumatic defects of the tibial plateau have provided a long-lasting and reliable reconstructive solution for a high demand population. Meniscal allografts should be used when clinically warranted. In the present study, all grafts were protected with a coincident realignment osteotomy when preoperative radiographs suggested that the allograft would be placed under increased load. Conversion to knee arthroplasty was required for approximately one-third of the patients at an average of ten years. PMID- 12721344 TI - What type of cartilage repair are we attempting to attain? PMID- 12721345 TI - Histological assessment of cartilage repair: a report by the Histology Endpoint Committee of the International Cartilage Repair Society (ICRS). PMID- 12721346 TI - Evaluation of cartilage injuries and repair. PMID- 12721347 TI - New MRI techniques for imaging cartilage. PMID- 12721349 TI - Surgical removal of articular cartilage leads to loss of chondrocytes from cartilage bordering the wound edge. AB - BACKGROUND: A number of arthroscopic procedures that are used in the treatment of focal cartilage lesions or osteoarthritic joints, such as shaving, debridement, and laser abrasion, involve the removal of both diseased and healthy articular cartilage. The excision of such tissue has the effect of generating lesions within the articular cartilage. The fate of the chondrocytes that border such lesions has not been evaluated. The purpose of this investigation was to ascertain whether the surgical creation of lesions in articular cartilage induces irreversible loss of chondrocytes over time from tissue bordering the wound edge and to determine whether the synthetic activity of cells in this region is compromised. METHODS: Partial-thickness defects of defined dimensions were created in the femoral condyle and/or trochlear groove of rabbits and miniature pigs. Cell volumes, cell volume densities, and numerical cell densities within tissue close to (within 100 micro m) and remote from (control site) the wound edge were determined by quantitative histomorphometry at various time intervals up to six months after surgery. Rates of proteoglycan synthesis by cells in both regions were determined by quantitative autoradiography following (35) S-sulphate labeling in vivo. RESULTS: The surgical creation of partial-thickness lesions in articular cartilage induced a significant and long-term loss of cells from tissue near the wound edge. However, the surviving cell population maintained a normal rate of matrix proteoglycan deposition. CONCLUSIONS: This study illustrates that maintenance and remodeling of cartilage matrix close to wound edges in articular cartilage lesions is compromised, since fewer cells, with an unchanged metabolic activity rate, are left to sustain matrix domains. PMID- 12721348 TI - Mechano-acoustic diagnosis of cartilage degeneration and repair. AB - BACKGROUND: The combined use of high-frequency ultrasound and mechanical indentation has been suggested for the evaluation of cartilage integrity. In this study, we investigated the usefulness of high-resolution B-mode ultrasound imaging and quantitative mechanical measurements for the diagnosis of cartilage degeneration and for monitoring tissue-healing after autologous chondrocyte transplantation. METHODS: In the first study, osteochondral samples (n = 32) were obtained from the lateral facet of a bovine patella, and the samples were visually classified as intact (n = 13) or degenerated (n = 19) and were graded with use of the Mankin scoring system. Samples were imaged with use of a 20-MHz ultrasound instrument, and the dynamic modulus (Edyn) of cartilage was determined in unconfined compression with use of a high-resolution materials tester. In the second study, cartilage chondrocytes were harvested from the low-weight-bearing area of six-month-old porcine knee joints and cultured. A month later, a cartilage lesion was created on the facet of the femoral trochlea and was repaired with use of the autologous chondrocyte transplantation technique (n = 10). Three months later, to estimate cartilage Edyn, the repair tissue, the adjacent cartilage, and the sham-operated contralateral joint cartilage (control) were analyzed in situ with an arthroscopic indentation instrument. Subsequently, the same sites were imaged with ultrasound. RESULTS: All visually degenerated bovine samples (mean Mankin score = 4) and five visually normal samples (Mankin score = 1) showed reduced Edyn (<2.1 MPa) as compared with histologically normal cartilage (Edyn = 13.8 +/- 3.2 MPa, Mankin score = 0). Cartilage stiffness, as shown by the indenter force, was lower (0.6 +/- 0.3 N, p < 0.05, Wilcoxon's signed-rank test) in the porcine tissue repaired with autologous chondrocyte transplantation than it was in the adjacent (1.6 +/- 0.1 N) or the control (1.9 +/- 0.4 N) tissue. The superficial and internal structure of the degenerated and repaired tissue, including the subchondral erosion at the repair site, was sensitively demonstrated by the ultrasound imaging. CONCLUSIONS: Measurement of cartilage Edyn is an objective method with which to follow changes in the mechanical integrity of cartilage. B-mode ultrasound imaging offers detailed information on the structural properties of cartilage and subchondral bone. PMID- 12721350 TI - Phenotypic plasticity of human articular chondrocytes. AB - BACKGROUND: Progenitor cells in mesenchymal tissues are important in the maintenance of tissue homeostasis and regeneration capacity. Articular cartilage is a tissue with a very low capacity for repair. One explanation could be the lack of chondrogenic progenitor cells within the adult tissue. As a test of chondrogenic differentiation potential, we examined the ability of isolated chondrocytes to take on several phenotypic identities within the mesenchymal lineage by applying culture techniques and markers used in the study of the phenotypic plasticity of marrow-derived mesenchymal stem cells (MSCs). METHODS: Culture-expanded human articular chondrocytes were analyzed for chondrogenic, adipogenic, and osteogenic capacity in defined in vitro culture systems. The osteochondrogenic potential of cells loaded into porous calcium-phosphate ceramic cubes implanted into mice was also determined. RESULTS: The different assays demonstrated that culture-expanded chondrocytes have the potential to form cartilage in pellet mass cultures, to form adipose cells in dense monolayer cultures, and to form a calcium-rich matrix in an osteogenic assay. In the in vitro assays, a variability of phenotypic plasticity was demonstrated among the donors. In contrast with MSCs, chondrocytes formed cartilage only (and not bone) in the in vivo osteochondrogenic assay. CONCLUSIONS: These results suggest that, within articular cartilage, there are chondrogenic cells that exhibit a level of phenotypic plasticity that is comparable with that of MSCs. However, there was a difference in the expression of bone in the in vivo assay. PMID- 12721351 TI - Effect of biomechanical conditioning on cartilaginous tissue formation in vitro. AB - BACKGROUND: Although tissue engineering of articular cartilage is a promising approach for cartilage repair, it has been difficult to develop cartilaginous tissue in vitro that mimics the properties of native cartilage. Isolated chondrocytes grown in culture typically do not accumulate enough extracellular matrix, and the generated tissue possesses only a fraction of the mechanical properties of native cartilage. One potential explanation for this might be that the cells are grown in an environment that lacks the mechanical stimuli to which the chondrocytes are exposed in vivo. In this study, we compared the long-term effects of both dynamic compressive and shearing forces on cartilaginous tissue formation in vitro. METHODS: Bovine articular chondrocytes were grown on the surface of porous ceramic substrates and were maintained under static, free swelling conditions for a period of four weeks. Cultures were then subjected to six minutes of mechanical stimulation every other day, in either compression or shear, for an additional four-week period. RESULTS: Cartilaginous tissues cultured in the presence of intermittent compression or shear were significantly thicker (p < 0.05) and had accumulated more extracellular matrix (p < 0.01) compared with the unstimulated controls. However, when normalized by the wet weight of the tissue, cultures stimulated in the presence of shearing forces contained more proteoglycans and collagen compared with compression-stimulated cultures. These cultures also displayed the largest increase in mechanical properties, with a threefold increase in equilibrium stress and a fivefold increase in equilibrium modulus. CONCLUSIONS AND CLINICAL RELEVANCE: The results of this study demonstrate that a brief application of mechanical forces applied periodically over a long duration can improve the quality of cartilaginous tissue formed in vitro. However, the changes in tissue composition and mechanical properties were dependent on the specific mode of the applied mechanical forces, with shear stimulation eliciting the greater effect. This finding suggests that chondrocytes may respond differently to different modes of applied forces. PMID- 12721352 TI - The role of chondrocyte senescence in the pathogenesis of osteoarthritis and in limiting cartilage repair. AB - BACKGROUND: With increasing age, the prevalence of osteoarthritis increases and the efficacy of articular cartilage repair decreases. As chondrocytes age, they synthesize smaller, less uniform aggrecan molecules and less functional link proteins, their mitotic and synthetic activity decline, and their responsiveness to anabolic mechanical stimuli and growth factors decreases. These observations led us to hypothesize that progressive cell senescence decreases the ability of chondrocytes to maintain and to restore articular cartilage. METHODS: To test this hypothesis, we measured cell senescence markers (beta-galactosidase expression, mitotic activity, and telomere length) in human articular cartilage chondrocytes from twenty-seven donors ranging in age from one to eighty-seven years. We also assessed mitochondrial DNA, membrane potential, and numerical density. To determine if chondrocyte age changes are reversible, we transfected human articular cartilage chondrocytes with the human telomerase gene (hTERT) and human papilloma virus oncogenes (E6 and E7). RESULTS: Beta-galactosidase expression increased with age (r = 0.84, p = 0.0001), while mitotic activity and telomere length declined (r = -0.77, p = 0.001 and r = -0.71, p = 0.0004, respectively). Decreasing telomere length was closely correlated with increasing expression of beta-galactosidase and decreasing mitotic activity. As the number of population doublings increased, mitochondrial DNA was degraded, mitochondrial membrane potential was lost, and the number of mitochondria per cell declined. Transfection of human articular cartilage chondrocytes from a forty-seven-year old donor with hTERT and human papilloma virus proto-oncogenes E6 and E7 created a cell line that has completed more than 300 population doublings as compared with an upper limit of twenty-five population doublings for normal cells. Telomere length increased in cells transduced with hTERT. CONCLUSIONS: These findings help to explain the previously reported age-related declines in chondrocyte synthetic activity, mitotic activity, and responsiveness to anabolic cytokines and mechanical stimuli. They also suggest that in vivo chondrocyte senescence contributes to the age-related increase in the prevalence of osteoarthritis and decrease in the efficacy of cartilage repair. The creation of immortal cells with increased telomere length suggests that the progression of human chondrocytes toward senescence is not inevitable. PMID- 12721353 TI - Stem cells as platforms for delivery of genes to enhance cartilage repair. AB - BACKGROUND: The long-held axiom put forth by Hunter in 1743, that cartilage once injured is incapable of healing, has been challenged by the technique of autologous chondrocyte transplantation. This conceptual change in the way in which orthopaedists are approaching the problem of cartilage repair has spawned a myriad of new and innovative treatment modalities. This review will focus on the new techniques and directions that our facility and other investigators are exploring to restore functional articular cartilage. METHODS: To show the usefulness and effectiveness of local tissue-engineered gene therapy, we transduced periosteal stem cells known to have osteochondral potential with either bone morphogenetic protein-7 (BMP-7) or sonic hedgehog (Shh) gene. These cells were cultured to increase the number of cells and then were seeded onto bioresorbable polymer scaffolds. Full-thickness osteochondral defects were created in the mid-trochlear region of eighty New Zealand White rabbits, and the implants containing the transduced cells were placed in the defects. Animals were killed at six, eight, twelve, and twenty-six weeks postoperatively and were examined macroscopically and histologically. RESULTS: Periosteal-derived cambium layer cells proliferated rapidly and were easily used for transfection of both the bone morphogenic protein-7 (BMP-7) and sonic hedgehog (Shh) genes. The control defects became filled with a mixture of fibrous and fibrocartilaginous tissue. The addition of either the BMP-7 or the Shh gene significantly enhanced the quality of the repair tissue, resulting in a much smoother surface and more hyaline-appearing cartilage. There was, however, a noticeable difference in the persistence of the cartilage phase between the group that received the Shh gene and the group that received the BMP-7 gene, with the subchondral compartment in the latter group seeming to remodel with bone much faster. CONCLUSION AND CLINICAL RELEVANCE: The results of these experiments clearly demonstrate the utility of tissue-engineering strategies in which gene therapy is used to locally influence the repair environment. It is interesting to note the relative differences in the two different gene responses with regard to skeletal development and the repair process. These differences could be related to the genes' temporal patterns in skeletal development. PMID- 12721355 TI - Molecular mechanisms regulating chondroblast differentiation. AB - BACKGROUND: Formation of the cartilage template involves a multi-step process in which prechondrogenic mesenchymal cells form condensations prior to differentiating into matrix-producing chondroblasts. Retinoids, particularly retinoic acid, are among the numerous signaling molecules that have been implicated in this process. A proper balance of retinoids is essential for normal skeletal development in that too much or too little negatively impacts skeletogenesis. During the past few years, substantial advances have been made in our understanding of the role of retinoid signaling in these processes, which is reviewed in this report. METHODS: To examine the function of retinoid signaling in skeletal development, transgenic mice that overexpressed a weak, constitutively active retinoic acid receptor (retinoic acid receptor-alpha) in their developing limbs were generated. The mice presented with a range of skeletal abnormalities. To examine the mechanisms responsible for these abnormalities, primary limb mesenchymal cultures from the transgenic mice were compared with cultures from wild-type mice. In addition, to address the molecular basis of retinoic acid receptor action, retinoic acid receptor activity in the primary cultures was manipulated with use of retinoic acid receptor-selective agonists and antagonists. The evaluation of the response to the manipulation of retinoic acid receptors was followed by histological studies and by the use of Northern blot analysis and reporter assays to analyze changes in the expression of chondrocytic markers and to monitor transcription factor activity, respectively. RESULTS: The evidence reviewed here indicates that retinoids maintain cells within condensations in a prechondrogenic, mesenchymal cell state, which prevents the cells from differentiating into chondroblasts. More recent studies have demonstrated that the inhibition of receptor-mediated retinoid signaling induces the expression of Sox9, a transcription factor that is considered a "master switch" for the differentiation of chondroblasts. These effects are largely mediated by the activation of the p38 MAPK signaling cascade. CONCLUSIONS: These findings demonstrate that retinoid receptor-mediated repression is both necessary and sufficient for chondroblast differentiation. Moreover, retinoic acid receptor repression acts downstream of BMP signaling or in a distinct pathway to activate p38 MAPK, which in turn induces chondroblast differentiation. PMID- 12721354 TI - Gene expression in chondrocytes assessed with use of microarrays. AB - BACKGROUND: Despite considerable limitations such as low sensitivity and insensitivity to alternative splicing, posttranscriptional regulation, and posttranslational modification, cDNA array technology provides a powerful tool with which to obtain an overview of gene expression patterns, hardly achievable with other techniques. This has been shown to be true for the analysis of known genes as well as the discovery of new genes of interest. METHODS: Samples of normal and late-stage osteoarthritic cartilage of human knee joints were analyzed with use of the Human Cancer 1.2 cDNA-array and TaqMan analysis. RESULTS: In spite of a large variability of expression levels among different patients, significant expression patterns for many known genes of interest such as cartilage matrix proteins (e.g., collagen types II, VI, and XI; aggrecan; decorin; biglycan) and matrix-degrading proteases were detected. Of the latter, MMP-3 appeared to be strongly expressed in normal and early degenerative cartilage and downregulated in the late disease stages. This indicates that, in the late stages of cartilage degeneration, other degradation pathways might be more important, for example, those involving enzymes such as MMP-2 and MMP-13, both of which were upregulated in late-stage disease. CONCLUSION: Most results have to be considered to be preliminary to a certain degree, as technical tools and interpretation approaches are still emerging and need more validation. Clearly, there is a major challenge to distill information and knowledge out of the obtained mass of data. However, these data will be one basis of a new world of biological understanding. These new insights will be network-based and no longer molecule-centered. Today, molecules have a biochemical and physiological context; tomorrow, biological networks will have molecules as constituents. PMID- 12721356 TI - The role of activating transcription factor-2 in skeletal growth control. PMID- 12721358 TI - A humble hexose monophosphate pathway metabolite regulates short- and long-term control of lipogenesis. PMID- 12721357 TI - Cellular signaling in developmental chondrogenesis: N-cadherin, Wnts, and BMP-2. PMID- 12721359 TI - The loss of the hemoglobin H2S-binding function in annelids from sulfide-free habitats reveals molecular adaptation driven by Darwinian positive selection. AB - The hemoglobin of the deep-sea hydrothermal vent vestimentiferan Riftia pachyptila (annelid) is able to bind toxic hydrogen sulfide (H(2)S) to free cysteine residues and to transport it to fuel endosymbiotic sulfide-oxidising bacteria. The cysteine residues are conserved key amino acids in annelid globins living in sulfide-rich environments, but are absent in annelid globins from sulfide-free environments. Synonymous and nonsynonymous substitution analysis from two different sets of orthologous annelid globin genes from sulfide rich and sulfide free environments have been performed to understand how the sulfide binding function of hemoglobin appeared and has been maintained during the course of evolution. This study reveals that the sites occupied by free-cysteine residues in annelids living in sulfide-rich environments and occupied by other amino acids in annelids from sulfide-free environments, have undergone positive selection in annelids from sulfide-free environments. We assumed that the high reactivity of cysteine residues became a disadvantage when H(2)S disappeared because free cysteines without their natural ligand had the capacity to interact with other blood components, disturb homeostasis, reduce fitness and thus could have been counterselected. To our knowledge, we pointed out for the first time a case of function loss driven by molecular adaptation rather than genetic drift. If constraint relaxation (H(2)S disappearance) led to the loss of the sulfide binding function in modern annelids from sulfide-free environments, our work suggests that adaptation to sulfide-rich environments is a plesiomorphic feature, and thus that the annelid ancestor could have emerged in a sulfide-rich environment. PMID- 12721360 TI - IL-17 production from activated T cells is required for the spontaneous development of destructive arthritis in mice deficient in IL-1 receptor antagonist. AB - IL-17 is a T cell-derived, proinflammatory cytokine that is suspected to be involved in the development of various inflammatory diseases. Although there are elevated levels of IL-17 in synovial fluid of patients with rheumatoid arthritis, the pathogenic role of IL-17 in the development of rheumatoid arthritis remains to be elucidated. In this report, the effects of IL-17 deficiency were examined in IL-1 receptor antagonist-deficient (IL-1Ra(-/-)) mice that spontaneously develop an inflammatory and destructive arthritis due to unopposed excess IL-1 signaling. IL-17 expression is greatly enhanced in IL-1Ra(-/-) mice, suggesting that IL-17 activity is involved in the pathogenesis of arthritis in these mice. Indeed, the spontaneous development of arthritis did not occur in IL-1Ra(-/-) mice also deficient in IL-17. The proliferative response of ovalbumin-specific T cells from DO11.10 mice against ovalbumin cocultured with antigen-presenting cells from either IL-1Ra(-/-) mice or wild-type mice was reduced by IL-17 deficiency, indicating insufficient T cell activation. Cross-linking OX40, a cosignaling molecule on CD4(+) T cells that plays an important role in T cell antigen-presenting cell interaction, with anti-OX40 Ab accelerated the production of IL-17 induced by CD3 stimulation. Because OX40 is induced by IL-1 signaling, IL-17 induction is likely to be downstream of IL-1 through activation of OX40. These observations suggest that IL-17 plays a crucial role in T cell activation, downstream of IL-1, causing the development of autoimmune arthritis. PMID- 12721361 TI - Targets of the transcriptional repressor oncoprotein Gfi-1. AB - Gfi-1 is a zinc finger transcriptional repressor originally recognized for its role in T cell differentiation and lymphomas. Recent experiments reveal that gene targeted Gfi-1-deficient mice are neutropenic and that Gfi-1 mutations cause human neutropenia. In both cases, myeloid progenitor cells lose the ability to distinctly differentiate granulocytes from monocytes. The molecular mechanism of the hematopoietic abnormalities caused by Gfi-1 deficiency remains undetermined because of a lack of known Gfi-1 target genes. To identify Gfi-1 targets in vivo, we performed large-scale chromatin immunoprecipitation analysis on a set of 34 candidate genes in myeloblast (KG-1 and HL-60), monoblast (U937), and T lymphocyte cell lines (Jurkat), in concert with RT-PCR-based expression profiling. We identified 32 Gfi-1 binding sites in a functionally variable set of 16 genes, including complements of cell-cycle regulators, transcription factors, and granulocyte-specific markers. Cluster analysis of expression patterns and chromatin immunoprecipitation data reveals that Gfi-1 targets a subset of genes differentiating hematopoietic lineages and therefore plays a relatively superior role in the hierarchy of factors governing stem cell differentiation. PMID- 12721362 TI - Tauroursodeoxycholic acid reduces apoptosis and protects against neurological injury after acute hemorrhagic stroke in rats. AB - Tauroursodeoxycholic acid (TUDCA), an endogenous bile acid, modulates cell death by interrupting classic pathways of apoptosis. Intracerebral hemorrhage (ICH) is a devastating acute neurological disorder, without effective treatment, in which a significant loss of neuronal cells is thought to occur by apoptosis. In this study, we evaluated whether TUDCA can reduce brain injury and improve neurological function after ICH in rats. Administration of TUDCA before or up to 6 h after stereotaxic collagenase injection into the striatum reduced lesion volumes at 2 days by as much as 50%. Apoptosis was approximately 50% decreased in the area immediately surrounding the hematoma and was associated with a similar inhibition of caspase activity. These changes were also associated with improved neurobehavioral deficits as assessed by rotational asymmetry, limb placement, and stepping ability. Furthermore, TUDCA treatment modulated expression of certain Bcl-2 family members, as well as NF-kappaB activity. In addition to its protective action at the mitochondrial membrane, TUDCA also activated the Akt 1protein kinase Balpha survival pathway and induced Bad phosphorylation at Ser 136. In conclusion, reduction of brain injury underlies the wide-range neuroprotective effects of TUDCA after ICH. Thus, given its clinical safety, TUDCA may provide a potentially useful treatment in patients with hemorrhagic stroke and perhaps other acute brain injuries associated with cell death by apoptosis. PMID- 12721363 TI - Linkage disequilibrium in human populations. AB - Whereas the human linkage map appears on limited evidence to be constant over populations, maps of linkage disequilibrium (LD) vary among populations that differ in gene history. The greatest difference is between populations of sub Saharan origin and populations remotely derived from Africa after a major bottleneck that reduced their heterozygosity and altered their Malecot parameters, increasing the intercept M that reflects association in founders and decreasing the exponential decline epsilon. Variation among populations within this ethnic dichotomy is much smaller. These observations validate use of a cosmopolitan LD map based on a sizeable sample representing a large population reliably typed for markers at high density. Then an LD map for a region or isolate within an ethnic group may be created by fitting the sample LD to the cosmopolitan map, estimating Malecot parameters simultaneously. The cosmopolitan map scaled by epsilon recovers 95% of the information that a local map at the same density gives and therefore more than the information in a low-resolution local map. Relative to a Eurasian cosmopolitan map the scaling factors are estimated to be 0.82 for isolates of European descent, 1.53 for Yorubans, and 1.74 for African Americans. These observations are consistent with a common bottleneck (perhaps but not necessarily speciation) approximately 173,500 years ago, if the bottleneck associated with migration out of Africa was 100,000 years ago. Eurasian populations (especially isolates with numerous cases) are efficient for genome scans, and populations of recent African origin (such as African Americans) are efficient for identification of causal polymorphisms within a candidate sequence. PMID- 12721364 TI - Catalytic and structural role of the metal ion in dUTP pyrophosphatase. AB - The metal ion dependence of the catalytic activity of recombinant Escherichia coli dUTP pyrophosphatase (dUTPase), an essential enzyme preventing incorporation of uracil into DNA, has been investigated by steady-state kinetic, electron paramagnetic resonance, and electron nuclear double resonance methods. Values of k(cat) and k(cat)K(m) were 4.5 +/- 0.1 s(-1) and 0.49 +/- 0.1 x 10(6) M(-1).s(-1) in the absence of divalent metal ions, 14.7 +/- 2.2 s(-1) and 25.1 +/- 7.4 x 10(6) M(-1).s(-1) in the presence of Mg(2+) or Mn(2+), and 24.2 +/- 3.6 s(-1) and 2.4 +/- 0.7 x 10(6) M(-1).s(-1) when supported by VO(2+) or bis(acetylacetonato)oxovanadium(IV). Binding of VO(2+) to the enzyme in the presence of dUDP, a nonhydrolyzable substrate analog, was specific and competitive with Mg(2+). Electron paramagnetic resonance spectra of the ternary enzyme-VO(2+)-chelate-dUDP complex revealed a pattern of (31)P superhyperfine coupling specifying two structurally equivalent phosphate groups equatorially coordinated to the VO(2+) ion. Proton electron nuclear double resonance spectra revealed an equatorial acetylacetonate ligand, indicating that one of the organic ligands had been displaced. By molecular graphics modeling, we show that the diphosphate group of enzyme-bound dUDP is sterically accessible to a hemi-chelate form of VO(2+). We propose a similar location compatible with all kinetic and spectroscopic results to account for the reactivity of VO(2+) and the VO(2+) chelate in dUTP hydrolysis. In this location the metal ion could promote an ordered conformation of the C-terminal fragment that is obligatory for catalysis but dynamically flexible in the free enzyme. PMID- 12721365 TI - Activation of CD4 T cells by Raf-independent effectors of Ras. AB - Small GTPase Ras is capable of mediating activation in T lymphocytes by using Raf kinase-dependent signaling pathway. Other effectors of Ras exist, however, suggesting that targets of Ras alternative to Raf may also contribute to T cell functions. Here we demonstrate that Ras(V12G37) mutant that fails to bind Raf, potently increases intracellular calcium concentration and cytokine production in primary antigen-stimulated T cells. From three known effectors which retain the ability to interact with Ras(V12G37), overexpression of phospholipase C epsilon but not that of RIN1 or Ral guanine nucleotide exchange factors enhanced cytokine and nuclear factor-activated T cell reporter T cell responses. Hence T cell activation can be critically regulated by the Ras effector pathway independent from Raf that can be mimicked by phospholipase C epsilon. PMID- 12721366 TI - SPARC-null mice exhibit increased adiposity without significant differences in overall body weight. AB - Secreted protein acidic and rich in cysteine/osteonectin/BM-40 (SPARC) is a matrix-associated protein that elicits changes in cell shape, inhibits cell-cycle progression, and influences the synthesis of extracellular matrix (ECM). The absence of SPARC in mice gives rise to aberrations in the structure and composition of the ECM that result in generation of cataracts, development of severe osteopenia, and accelerated closure of dermal wounds. In this report we show that SPARC-null mice have greater deposits of s.c. fat and larger epididymal fat pads in comparison with wild-type mice. Similar to earlier studies of SPARC null dermis, we observed a reduction in collagen I in SPARC-null fat pads in comparison with wild-type. Although elevated levels of serum leptin were observed in SPARC-null mice, their overall body weights were not significantly different from those of wild-type counterparts. The diameters of adipocytes from SPARC-null versus wild-type epididymal fat pads were 252 +/- 61 and 161 +/- 33 microm (means +/- SD), respectively, and there was an increase in adipocyte number within SPARC null fat pads in comparison with wild-type pads. Thus the absence of SPARC appears to result in an increase in the size of individual adipocytes as well as an increase in the number of adipocytes per fat pad. In fat pads isolated from wild-type mice, SPARC mRNA was associated with both the stromal/vascular and adipocyte fractions. We propose that SPARC limits the accumulation of adipose tissue in mice in part through its demonstrated effects on the regulation of cell shape and production of ECM. PMID- 12721367 TI - Dynamic assembly of surface structures in living cells. AB - Although the dynamics of cell membranes and associated structures is vital for cell function, little is known due to lack of suitable methods. We found, using scanning ion conductance microscopy, that microvilli, membrane projections supported by internal actin bundles, undergo a life cycle: fast height-dependent growth, relatively short steady state, and slow height-independent retraction. The microvilli can aggregate into relatively stable structures where the steady state is extended. We suggest that the intrinsic dynamics of microvilli, combined with their ability to make stable structures, allows them to act as elementary "building blocks" for the assembly of specialized structures on the cell surface. PMID- 12721368 TI - The mechanism of ferrichrome transport through Arn1p and its metabolism in Saccharomyces cerevisiae. AB - Siderophores are low molecular weight compounds, synthesized and secreted by microorganisms, that specifically bind ferric iron with exceptionally high affinity. Microbes capture these compounds and take up the bound iron through specific, high-affinity systems. Saccharomyces cerevisiae can take up iron bound to siderophores through the transporters of the ARN family; however, the mechanism by which the siderophore-bound iron enters the cell via these transporters is not known. Here we describe how ferrichrome, a siderophore of the hydroxamate class, is taken up by Arn1p. Arn1p exhibits two surface binding sites for ferrichrome, one that is similar in affinity to the K(T) for uptake and one of a much higher affinity that is specific for the metallated form of ferrichrome. Ferrichrome may gain access to the higher-affinity site through endocytosis. Tracer studies using (14)C-labeled ferrichrome bound to either iron(III) or aluminum(III), a nonreducible ligand for ferrichrome, indicate that ferrichrome enters the cell as the intact metallosiderophore and accumulates in the cytosol. Both ferrichrome chelates were relatively stable within the cell, and metal-free ferrichrome did not accumulate, indicating a role for ferrichrome in intracellular iron storage. Iron stored as ferrichrome was readily mobilized to meet the metabolic needs of the cell. PMID- 12721369 TI - Folding quality control in the export of proteins by the bacterial twin-arginine translocation pathway. AB - To examine the relationship between folding and export competence by the twin arginine translocation (Tat) pathway we analyzed the subcellular localization of fusions between a set of eight putative Tat leader peptides and alkaline phosphatase in isogenic Escherichia coli strains that either allow or disfavor the formation of protein disulfide bonds in the cytoplasm. We show that export by the Tat translocator is observed only in strains that enable oxidative protein folding in the cytoplasm. Further, we show that other disulfide-containing proteins, namely single-chain Fv and heterodimeric F(AB) antibody fragments, are export-competent only in strains having an oxidizing cytoplasm. Functional, heterodimeric F(AB) protein was exported from the cytoplasm by means of a Tat leader peptide fused to the heavy chain alone, indicating that the formation of a disulfide-bonded dimer preceeds export. These results demonstrate that in vivo only proteins that have attained the native conformation are exported by the Tat translocator, indicating that a folding quality-control mechanism is intrinsic to the export process. The ability to export proteins with disulfide bonds and the folding proofing feature of the Tat pathway are of interest for biotechnology applications. PMID- 12721370 TI - NADPH oxidase mediates oxidative stress in the 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine model of Parkinson's disease. AB - Parkinson's disease (PD) is a neurodegenerative disorder of uncertain pathogenesis characterized by a loss of substantia nigra pars compacta (SNpc) dopaminergic (DA) neurons, and can be modeled by the neurotoxin 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine (MPTP). Both inflammatory processes and oxidative stress may contribute to MPTP- and PD-related neurodegeneration. However, whether inflammation may cause oxidative damage in MPTP and PD is unknown. Here we show that NADPH-oxidase, the main reactive oxygen species (ROS)-producing enzyme during inflammation, is up-regulated in SNpc of human PD and MPTP mice. These changes coincide with the local production of ROS, microglial activation, and DA neuronal loss seen after MPTP injections. Mutant mice defective in NADPH-oxidase exhibit less SNpc DA neuronal loss and protein oxidation than their WT littermates after MPTP injections. We show that extracellular ROS are a main determinant in inflammation-mediated DA neurotoxicity in the MPTP model of PD. This study supports a critical role for NADPH-oxidase in the pathogenesis of PD and suggests that targeting this enzyme or enhancing extracellular antioxidants may provide novel therapies for PD. PMID- 12721372 TI - Acetylcholine enhancement in the nucleus accumbens prevents addictive behaviors of cocaine and morphine. AB - Drug addiction poses serious social, medical, and economic problems, but effective treatments for drug addiction are still limited. Cocaine and morphine elevate dopamine levels in the nucleus accumbens (NAc), and the overwhelming actions of dopamine are implicated in reinforcement and addiction of abusive drugs. In our previous studies, we reported the regulatory role of acetylcholine (ACh) in the NAc function by selectively ablating the NAc cholinergic neurons with use of immunotoxin-mediated cell targeting. These studies indicated that ACh and dopamine acted convergently but oppositely on the NAc circuit and that cholinergic cell ablation enhanced long-lasting behavioral changes of cocaine addiction. In this investigation, we showed that immunotoxin-mediated ablation of the NAc cholinergic neurons enhanced not only the sensitivity to morphine in conditioned place preference but also negative reinforcement of morphine withdrawal in conditioned place aversion. Remarkably, acetylcholinesterase (AChE) inhibitors that act on the brain AChE suppressed both cocaine- and morphine induced conditioned place preference and blocked the induction and persistence of cocaine-evoked hyperlocomotion. Importantly, this inhibition was abolished by ablation of the NAc cholinergic neurons. These results demonstrate that centrally active AChE inhibitors prevent long-lasting behavioral abnormalities associated with cocaine and morphine addictions by potentiating the actions of ACh released from the NAc cholinergic neurons. Centrally active AChE inhibitors could thus be approached as novel and potential therapeutic agents for drug addiction. PMID- 12721371 TI - Transgenic overexpression of galanin in the dorsal root ganglia modulates pain related behavior. AB - The neuropeptide galanin is expressed in the dorsal root ganglia (DRG) and spinal cord and is thought to be involved in the modulation of pain processing. However, its mechanisms of action are complex and poorly understood, as both facilitatory and inhibitory effects have been described. To understand further the role played by galanin in nociception, we have generated two transgenic lines that overexpress galanin in specific populations of primary afferent DRG neurons in either an inducible or constitutive manner. In the first line, a previously defined enhancer region from the galanin locus was used to target galanin to the DRG (Gal-OE). Transgene expression recapitulates the spatial endogenous galanin distribution pattern in DRG neurons and markedly overexpresses the peptide in the DRG after nerve injury but not in the uninjured state. In the second line, an enhancer region of the c-Ret gene was used to constitutively and ectopically target galanin overexpression to the DRG (Ret-OE). The expression of this second transgene does not alter significantly after nerve injury. Here, we report that intact Ret-OE, but not Gal-OE, animals have significantly elevated mechanical and thermal thresholds. After nerve damage, using a spared nerve-injury model, mechanical allodynia is attenuated markedly in both the Gal-OE and Ret-OE mice compared with WT controls. These results support an inhibitory role for galanin in the modulation of nociception both in intact animals and in neuropathic pain states. PMID- 12721373 TI - Mutational analysis of predicted interactions between the catalytic and P domains of prohormone convertase 3 (PC3/PC1). AB - The subtilisin-like prohormone convertases (PCs) contain an essential downstream domain (P domain), which has been predicted to have a beta-barrel structure that interacts with and stabilizes the catalytic domain (CAT). To assess possible sites of hydrophobic interaction, a series of mutant PC3-enhanced GFP constructs were prepared in which selected nonpolar residues on the surface of CAT were substituted by the corresponding polar residues in subtilisin Carlsberg. To investigate the folding potential of the isolated P domain, signal peptide-P domain-enhanced GFP constructs with mutated andor truncated P domains were also made. All mutants were expressed in betaTC3 cells, and their subcellular localization and secretion were determined. The mutants fell into three main groups: (i) Golgisecreted, (ii) ERnonsecreted, and (iii) apoptosis inducing. The destabilizing CAT mutations indicate that the side chains of V292, T328, L351, Q408, H409, V412, and F441 and nonpolar fragments of the side chains of R405 and W413 form a hydrophobic patch on CAT that interacts with the P domain. We also have found that the P domain can fold independently, as indicated by its secretion. Interestingly, T594, which is near the P domain C terminus, was not essential for P domain secretion but is crucial for the stability of intact PC3. T594V produced a stable enzyme, but T594D did not, which suggests that T594 participates in important hydrophobic interactions within PC3. These findings support our conclusion that the catalytic and P domains contribute to the folding and thermodynamic stability of the convertases through reciprocal hydrophobic interactions. PMID- 12721374 TI - The rate of hydrolysis of phosphomonoester dianions and the exceptional catalytic proficiencies of protein and inositol phosphatases. AB - To evaluate the proficiency of phosphatases as catalysts, the rate of the uncatalyzed hydrolysis of simple phosphate monoester dianions was estimated by extrapolating rates measured over a range of high temperatures. The rate of spontaneous hydrolysis of phenyl phosphate dianion indicates that a linear free energy relationship reported earlier is reliable for leaving groups whose conjugate acids have pKa values up to at least 10. Using Teflon reaction vessels, it proved possible to follow the hydrolysis of methyl phosphate and 3-(4-carboxy) 2,2-dimethylpropyl phosphate in strong alkali. Even in 1 M KOH, the reaction was found to be specific acid catalyzed. These results establish an upper limit for dianion reactivity, which had been overestimated earlier as a result of the leaching by alkali of silicic acid from quartz reaction vessels. The present findings indicate that the half-time for attack by water on alkyl phosphate dianions is 1.1 x 10(12) years (k = 2 x 10(-20) s) at 25 degrees C and that phosphatases involved in cell signaling and regulation produce the largest rate enhancements that have been identified thus far. Protein phosphatase-1 and inositol 1-phosphatase exceed all other known enzymes in their affinities for the altered substrates in the transition state. PMID- 12721377 TI - Temporal association of Ca(2+)-dependent protein kinase with oil bodies during seed development in Santalum album L.: its biochemical characterization and significance. AB - Calcium-dependent protein kinase (CDPK) is expressed in sandalwood (Santalum album L.) seeds under developmental regulation, and it is localized with spherical storage organelles in the endosperm [Anil et al. (2000) Plant Physiol. 122: 1035]. This study identifies these storage organelles as oil bodies. A 55 kDa protein associated with isolated oil bodies, showed Ca(2+)-dependent autophosphorylation and also cross-reacted with anti-soybean CDPK. The CDPK activity detected in the oil body-protein fraction was calmodulin-independent and sensitive to W7 (N-(6-aminohexyl)-5-chloro-1-naphthalene sulfonamide) inhibition. Differences in Michaelis Menton kinetics, rate of histone phosphorylation and sensitivity to W7 inhibition between a soluble CDPK from embryos and the oil body associated CDPK of endosperm suggest that these are tissue-specific isozymes. The association of CDPK with oil bodies of endosperm was found to show a temporal pattern during seed development. CDPK protein and activity, and the in vivo phosphorylation of Ser and Thr residues were detected strongly in the oil bodies of endosperm from maturing seed. Since oil body formation occurs during seed maturation, the observations indicate that CDPK and Ca(2+) may have a regulatory role during oil accumulation/oil body biogenesis. The detection of CDPK-protein and activity in oil bodies of groundnut, sesame, cotton, sunflower, soybean and safflower suggests the ubiquity of the association of CDPKs with oil bodies. PMID- 12721375 TI - Resistance of cell membranes to different detergents. AB - Partial resistance of cell membranes to solubilization with mild detergents and the analysis of isolated detergent-resistant membranes (DRMs) have been used operationally to define membrane domains. Given the multitude of detergents used for this purpose, we sought to investigate whether extraction with different detergents might reflect the same underlying principle of domain formation. We therefore compared the protein and lipid content of DRMs prepared with a variety of detergents from two cell lines. We found that the detergents differ considerably in their ability to selectively solubilize membrane proteins and to enrich sphingolipids and cholesterol over glycerophospholipids as well as saturated over unsaturated phosphatidylcholine. In addition, we observed cell type-dependent variations of the molecular characteristics of DRMs and the effectiveness of particular detergents. These results make it unlikely that different detergents reflect the same aspects of membrane organization and underscore both the structural complexity of cell membranes and the need for more sophisticated analytical tools to understand their architecture. PMID- 12721378 TI - The cDNA microarray analysis using an Arabidopsis pad3 mutant reveals the expression profiles and classification of genes induced by Alternaria brassicicola attack. AB - The hypersensitive response (HR) was induced in a wild-type Arabidopsis thaliana plant (Columbia) (Col-wt) by inoculation with Alternaria brassicicola that causes the development of small brown necrotic lesions on the leaves. By contrast, pad3 1 mutants challenged with A. brassicicola produced spreading lesions. The cell death in pad3-1 mutants could not inhibit the pathogen growth and development, although both production of H(2)O(2) and localized cell death were similar in Col wt and pad3-1 plants after the inoculation. The difference between Col-wt and pad3-1 plants is defense responses after the occurrence of cell death. In other words, PAD3 is necessary for defense response to A. brassicicola. Therefore, we examined the changes in the expression patterns of ca. 7,000 genes by cDNA microarray analysis after inoculation with A. brassicicola. The cDNA microarrays were also done to analyze Arabidopsis responses after treatment with signal molecules, reactive oxygen species (ROS)-inducing compounds and UV-C. The results suggested that the pad3-1 mutation altered not only the accumulation of camalexin but also the timing of expression of many defense-related genes in response to the challenge with A. brassicicola. Furthermore, the plants integrate two or more signals that act together for promoting the induction of multiple defense pathways. PMID- 12721379 TI - Enzymatic and non-enzymatic antioxidant responses of Carrizo citrange, a salt sensitive citrus rootstock, to different levels of salinity. AB - In several plant species, oxidative stress has been shown to be one of the causes of damage produced by salinity. In order to assess the implication of oxidative stress in the reported sensitivity of the citrus rootstock Carrizo citrange to salt stress, 5-month-old seedlings were grown with increasing NaCl concentrations added to the watering solution. As an indicator of oxidative damage, malondialdehyde content was measured. The antioxidant capability of the plants was determined by measuring superoxide dismutase, ascorbate peroxidase, catalase and glutathione reductase activities together with the non-enzymatic antioxidant activity. As additional physiological responses to the stress, 1 aminocyclopropane-1-carboxilic acid and proline accumulation were assessed. Data indicate that Carrizo citrange responded to salt-induced oxidative stress by increasing enzymatic and non-enzymatic antioxidant defenses proportionally to the extent of the stress imposed, and that in all plants the malondialdehyde content remained at a moderate level. We suggest that the important deleterious effects reported in Carrizo citrange grown under high NaCl concentrations are mainly due to a cellular intoxication by Cl(-) ions and not to the salt-induced oxidative stress. PMID- 12721380 TI - Camptothecin biosynthetic genes in hairy roots of Ophiorrhiza pumila: cloning, characterization and differential expression in tissues and by stress compounds. AB - Camptothecin derivatives are clinically used anti-tumor compounds that biogenetically belong to a group of monoterpenoid indole alkaloids (TIA). We have already established a hairy root culture of Ophiorrhiza pumila (Rubiaceae) that produces camptothecin. The present study describes the cloning and characterization of cDNAs encoding strictosidine synthase (OpSTR; EC 4.3.3.2) and tryptophan decarboxylase (OpTDC; EC 4.1.1.28), two key enzymes in the biosynthesis of TIA from hairy roots of O. pumila. We also isolated the cDNA coding for NADPH:cytochrome P450 reductase (OpCPR; EC 1.6.2.4) that is presumed to be indirectly involved in camptothecin synthesis. The recombinant OpSTR and OpTDC proteins exhibit STR and TDC activities, respectively, when expressed in Escherichia coli. The tissue-specific and stress-inducible expression patterns of OpSTR and OpTDC were quite similar, unlike those of OpCPR. The high expression of OpSTR and OpTDC observed in hairy roots, roots and stems were closely correlated with STR protein accumulation as observed by immunoblot analysis. Plant stress compounds like salicylic acid repressed expression of OpSTR and OpTDC, suggesting coordinate regulation of these genes for camptothecin biosynthesis. PMID- 12721381 TI - Up-regulation of soyasaponin biosynthesis by methyl jasmonate in cultured cells of Glycyrrhiza glabra. AB - Exogenously applied methyl jasmonate (MeJA) stimulated soyasaponin biosynthesis in cultured cells of Glycyrrhiza glabra (common licorice). mRNA level and enzyme activity of beta-amyrin synthase (bAS), an oxidosqualene cyclase (OSC) situated at the branching point for oleanane-type triterpene saponin biosynthesis, were up regulated by MeJA, whereas those of cycloartenol synthase, an OSC involved in sterol biosynthesis, were relatively constant. Two mRNAs of squalene synthase (SQS), an enzyme common to both triterpene and sterol biosyntheses, were also up regulated by MeJA. In addition, enzyme activity of UDP-glucuronic acid: soyasapogenol B glucuronosyltransferase, an enzyme situated at a later step of soyasaponin biosynthesis, was also up-regulated by MeJA. Accumulations of bAS and two SQS mRNAs were not transient but lasted for 7 d after exposure to MeJA, resulting in the high-level accumulation (more than 2% of dry weight cells) of soyasaponins in cultured licorice cells. In contrast, bAS and SQS mRNAs were coordinately down-regulated by yeast extract, and mRNA accumulation of polyketide reductase, an enzyme involved in 5-deoxyflavonoid biosynthesis in cultured licorice cells, was induced transiently by yeast extract and MeJA, respectively. PMID- 12721382 TI - Isolation of stress-related genes of rubber particles and latex in fig tree (Ficus carica) and their expressions by abiotic stress or plant hormone treatments. AB - Two rubber particle protein genes and one latex gene in fig tree (Ficus carica) have been isolated and their expression following various abiotic stress treatments have been investigated. The two major proteins that are tightly associated with the catalytically active rubber particles have been sequenced to be peroxidase (POX) and trypsin inhibitor (TRI). A cDNA encoding a basic class I chitinase (CHI) has also been isolated from the fig tree latex. Wounding treatment strongly induced the expression of the three stress-related genes. Among the abiotic stresses investigated, drought treatment greatly induced the expression of POX, whereas the expression of CHI and TRI decreased after the same treatment. Cold treatment reduced slightly the transcript levels of the thee genes, and NaCl reduced marginally the expression of CHI. The expression of POX, CHI, and TRI was induced by jasmonic acid and abscisic acid, by jasmonic acid, and by salicylic acid, respectively. Different expression of the stress-related genes following various abiotic stress or plant hormone treatments suggests that a crosstalk exists between the signal transduction pathways elicited by abiotic stresses and hormones in plants. Our present results showing the expression of stress-related proteins on the surface of rubber particles and latex in F. carica also imply the possible role of rubber particles and latex in defense in rubber producing plant species. PMID- 12721383 TI - Time course analysis of apoptotic cell death during expression of hybrid lethality in hybrid tobacco cells (Nicotiana suaveolens x N. tabacum). AB - Hybrid cells from the cross Nicotiana suaveolens x N. tabacum expressed hybrid lethality at 28 degrees C in a thin layer cell culture system. Features characteristic of apoptosis, such as DNA fragmentation, chromatin condensation and nuclear fragmentation, were detected during expression of hybrid lethality. Actinomycin D (ActD) or cycloheximide (CHX) added to the medium suppressed apoptotic cell death during hybrid lethality. This indicates that hybrid lethality requires de novo transcription and translation, and is thus under genetic control. To estimate the time course of apoptotic cell death during the expression of hybrid lethality, we determined when factors controlling hybrid lethality were expressed by observing the point of no return. When cells were exposed to 28 degrees C for 2 h or less in inhibitor-free medium before addition of ActD or CHX, the percentage of dead cells did not increase. However, when cells were exposed to 28 degrees C for 4 h before the addition of inhibitor, the percentage of dead cells increased. When cells were exposed to 28 degrees C for 3 h before the addition of inhibitor, the percentage of dead cells varied from experiment to experiment. These data indicate that the factors controlling hybrid lethality are expressed 3 h after induction of hybrid lethality. In addition, we found a time difference between the expression of cell death and nuclear fragmentation. This suggests that the factor controlling cell death is different from the one controlling nuclear fragmentation. PMID- 12721384 TI - Immunohistochemical studies on translocation of pollen S-haplotype determinant in self-incompatibility of Brassica rapa. AB - The self-incompatibility system in Brassica is controlled by the S-locus, which contains S-receptor kinase (SRK) and S-locus protein 11 (SP11). SRK and SP11 control stigma and pollen S-haplotype specificity, respectively. SP11 binding to SRK induces the autophosphorylation of SRK, which triggers the signaling cascade that results in the rejection of self-pollen. The localization of SP11 protein during pollen development and pollination, however, have never been demonstrated. In this study, we examined the localization of S(8)-SP11 protein in the anther or pollinated stigma by immuno-electron microscopy. The immunostaining suggested that S(8)-SP11 was secreted from the tapetal cell into the anther locule as a cluster and translocated to the pollen surface at the early developmental stage of the anther. During the pollination process, SP11 was translocated from the pollen surface to the papilla cell, and then penetrated the cuticle layer of the papilla cell to diffuse across the pectin cellulose layer. Furthermore, SP11 protein could only penetrate the cuticle layer of the papilla cell in the presence of pollen grains, and could not penetrate on its own. This suggests that another factor from the pollen grain is needed for SP11 protein to penetrate the papilla cell wall. PMID- 12721385 TI - Molecular cloning and characterization of a cDNA encoding a novel apoplastic protein preferentially expressed in a shikonin-producing callus strain of Lithospermum erythrorhizon. AB - A cDNA (LEPS-2) encoding a novel cell wall protein was cloned from shikonin producing callus tissues of Lithospermum erythrorhizon by differential display between a shikonin-producing culture strain and a non-producing strain. The LEPS 2 cDNA encoded a polypeptide of 184 amino acids. The deduced amino acid sequence exhibited no significant homology with known proteins. Expression of LEPS-2 gene as well as accumulation of LEPS-2 protein was highly correlated with shikonin production in L. erythrorhizon cells in culture. In the intact plant, expression of LEPS-2 was detected only in the roots where shikonin pigments accumulated. Cell fractionation experiments and immunocytochemical analysis showed that the protein was localized in the apoplast fraction of the cell walls. The shikonin pigments were also stored on the cell walls as oil droplets. These results indicate that expression of the LEPS-2 is closely linked with shikonin biosynthesis and the LEPS-2 protein may be involved in the intra-cell wall trapping of shikonin pigments. PMID- 12721386 TI - Transformation of tobacco with a gene for the thermophilic acyl-lipid desaturase enhances the chilling tolerance of plants. AB - The desC gene for the acyl-lipid Delta9-desaturase from the thermophilic cyanobacterium Synechococcus vulcanus was introduced into Nicotiana tabacum under control of the 35S promoter. Expression of the desaturase was confirmed by Western blotting. Lipid analysis revealed that lipid content and the extent of fatty acid unsaturation significantly increased in leaves of transgenic plants. Chilling tolerance of those plants also increased, as estimated by the electrolyte leakage from the tissues damaged by cold treatments. Seeds of plants that expressed the desC gene imbibed at low temperatures demonstrated higher chilling tolerance than those of the control plants. The results demonstrate that the cyanobacterial thermophilic acyl-lipid desaturase was efficiently expressed in tobacco at ambient temperatures, and its expression resulted in the enhanced chilling tolerance of the transgenic plants. PMID- 12721387 TI - Dynamic Interaction between the D1 protein, CP43 and OEC33 at the lumenal side of photosystem II in spinach chloroplasts: evidence from light-induced cross-Linking of the proteins in the donor-side photoinhibition. AB - During the donor-side photoinhibition of spinach photosystem II, the reaction center D1 protein cross-linked with the antenna chlorophyll binding protein CP43 of photosystem II lacking the oxygen-evolving complex (OEC) subunit proteins. The cross-linking did not occur upon illumination of photosystem II samples that retained the OEC33, nor when OEC33-depleted photosystem II samples were reconstituted with the OEC33 prior to illumination. These results suggest that the D1 protein, CP43 and the OEC33 are located in close proximity at the lumenal side of photosystem II, and that the OEC33 suppresses the unnecessary contact between the D1 protein and CP43. Previously we presented data showing the D1 protein located adjacent to CP43 on the stromal side of photosystem II [Ishikawa et al. (1999) BIOCHIM: Biophys. Acta 1413: 147]. The present data suggest that the spatial arrangement of the D1 protein and CP43 at the lumenal side of photosystem II in spinach chloroplasts is similar to that at the stromal side of photosystem II and is consistent with the assignment of these proteins recently proposed on the crystal structures of the photosystem II complexes from cyanobacteria [Zouni et al. (2001) Nature 409: 739, Kamiya and Shen 2003 PROC: Natl. Acad. Sci. USA, 100: 98]. Moreover, the data suggest that the binding condition and positioning of the OEC33 in the photosystem II complex from higher plants may be different from those in cyanobacteria. PMID- 12721388 TI - Cyclic electron flow within PSII protects PSII from its photoinhibition in thylakoid membranes from spinach chloroplasts. AB - Cyclic electron flow within PSII (CEF-PSII) was proven to alleviate the photoinhibition of PSII. We set the conditions where CEF-PSII functioned or did not, by adding nigericin to the reaction mixture for the dissipation of DeltapH across thylakoid membranes, and then the thylakoids were illuminated. When CEF PSII did not function and the activity of linear electron flow (LEF) was low, light-treated thylakoid membranes largely lost the activity of LEF. The inactivation of LEF was due to the loss of the activity of PSII, but not that of PSI. The inactivation of PSII was suppressed, when CEF-PSII functioned or LEF was enhanced. These results imply that CEF-PSII contributes to the protection of PSII from its photoinhibition with LEF, as an electron sink. PMID- 12721389 TI - Calcium and the failing heart: phospholamban, good guy or bad guy? AB - In cardiac cells, phospholamban is a potent inhibitor of sarcoplasmic reticulum calcium (Ca(2+)) transport. Overexpression of mutant forms of phospholamban may result in beneficial or detrimental effects on intracellular Ca(2+) handling and cardiac systolic and diastolic function. Mutations in phospholamban have also been linked to human cardiomyopathies, providing important insights into the underlying disease mechanisms and the key role of phospholamban in myocardial excitation-contraction coupling. This Perspective discusses new advances in our understanding of the role of phospholamban in intracellular Ca(2+) handling and the development of human and murine cardiomyopathies. PMID- 12721390 TI - Capsaicinoids cause inflammation and epithelial cell death through activation of vanilloid receptors. AB - Capsaicinoids, found in less-than-lethal self-defense weapons, have been associated with respiratory failure and death in exposed animals and people. The studies described herein provide evidence for acute respiratory inflammation and damage to epithelial cells in experimental animals, and provide precise molecular mechanisms that mediate these effects using human bronchiolar and alveolar epithelial cells. Inhalation exposure of rats to pepper sprays (capsaicinoids) produced acute inflammation and damage to nasal, tracheal, bronchiolar, and alveolar cells in a dose-related manner. In vitro cytotoxicity assays demonstrated that cultured human lung cells (BEAS-2B and A549) were more susceptible to necrotic cell death than liver (HepG2) cells. Transcription of the human vanilloid receptor type-1, VR1 or TRPV1, was demonstrated by RT-PCR in all of these cells, and the relative transcript levels were correlated to cellular susceptibility. TRPV1 receptor activation was presumably responsible for cellular cytotoxicity, but prototypical functional antagonists of this receptor were cytotoxic themselves, and did not ameliorate capsaicinoid-induced damage. Conversely, the TRPV1 antagonist capsazepine, as well as calcium chelation by EGTA ablated cytokine (IL-6) production after capsaicin exposure. To address these seemingly contradictory results, recombinant human TRPV1 was cloned and overexpressed in BEAS-2B cells. These cells exhibited dramatically increased cellular susceptibility to capsaicinoids, measured using IL-6 production and cytotoxicity, and an apoptotic mechanism of cell death. Surprisingly, the cytotoxic effects of capsaicin in TRPV1 overexpressing cells were also not inhibited by TRPV1 antagonists or by treatments that modified extracellular calcium. Thus, capsaicin interacted with TRPV1 expressed by BEAS-2B and other airway epithelial cells to cause the calcium-dependent production of cytokines and, conversely, calcium-independent cell death. These results have demonstrated that capsaicinoids contained in pepper spray products produce airway inflammation and cause respiratory epithelial cell death. The mechanisms of these cellular responses to capsaicinoids appear to proceed via distinct cellular pathways, but both pathways are initiated by TRPV1. PMID- 12721393 TI - Hypersensitivity reactions to non-beta-lactam antibiotics. AB - As alternatives to beta lactam antibiotics continue to be developed, an increasing number of patients are experiencing hypersensitivity reactions to these agents. Unfortunately, unlike penicillin, there are no validated skin testing reagents to aid in confirming the presence of specific IgE antibodies to these drugs. This review summarizes what is known about hypersensitivity reactions to non-beta lactam antibiotics, and includes practical approaches to readministering these drugs in selected situations. Sulfonamides, vancomycin, macrolides, tetracyclines, fluoroquinolones, and anti-virals are included in the discussion. PMID- 12721392 TI - Hypersensitivity reactions to beta-lactam antibiotics. AB - Clinicians commonly encounter patients with a history of allergy to penicillin and other beta-lactam antibiotics, since about 10% of the population reports such an allergy. At the same time, it is known that about 90% of these patients are not truly allergic and could safely receive beta-lactam antibiotics. Instead, these patients are treated unnecessarily with alternate broad-spectrum antibiotics, which increases costs and contributes to the development and spread of multiple drug-resistant bacteria. In the case of penicillin, relevant allergenic determinants that elicit immune responses are known. Hence, validated diagnostic skin testing to detect the presence of drug-specific IgE antibodies is available. For non-penicillin beta-lactams, the immunogenic determinants that are produced by degradation are unknown, and diagnostic skin testing is of more limited value. Ideally, patients with a history of penicillin allergy should be evaluated when they are well and not in immediate need of antibiotic therapy. Patients who are found to be penicillin skin test-negative may be safely treated with all beta-lactam antibiotics. Penicillin skin test-positive patients should only receive a penicillin-class antibiotic via rapid desensitization, and only in cases when an alternative agent cannot be substituted. Penicillin skin test positive patients may be safely treated with monobactams. The extent of allergic cross-reactivity between penicillin arid cephalosporins/carbapenems is uncertain; therefore penicillin skin test-positive patients should only receive these antibiotics via cautious graded challenge or desensitization. Identification of patients who erroneously carry a label of beta-lactam allergy leads to improved utilization of antibiotics and slows the spread of multiple drug-resistant bacteria. PMID- 12721394 TI - Reactive metabolites and adverse drug reactions: clinical considerations. AB - Idiosyncratic reactions can affect many different organ systems, either as -an isolated event (e.g., hepatitis) or as part of a syndrome (e.g., drug hypersensitivity syndrome). Formation of reactive metabolites of drugs in conjunction with a decreased ability for detoxification is believed to be the initiating step in many idiosyncratic reactions. The drug hypersensitivity syndrome, defined by the presence of fever, rash and internal organ involvement, is estimated to occur in approx 1 in 1000 to 1 in 10,000 exposures with drugs such as anticonvulsants sulfonamide antibiotics, allopurinol, and dapsone. Symptoms usually start within 2-8 wk of drug initiation. Serum sicknesslike reaction, most frequently found after 7-14 d of drug exposure, is distinguished by a fever, cutaneous eruption and arthralgias. Medications such as cefaclor, minocycline and bupropion are most frequently implicated in this reaction. In contrast, drug-induced lupus can occur l-2 yr after initiation of medication. Drug-induced lupus is characterized by musculoskeletal complaints and fever and weight loss. Drugs most commonly associated with drug-induced lupus include procainamide, hydralazine, chlorpromazine, isoniazid, and minocycline. Management of patients who develop idiosyncratic reactions includes discontinuation of the implicated drug, initiation of corticosteroids (when appropriate), and symptomatic relief as required. Internal organ involvement, which may initially be asymptomatic, should be monitored. Confirmatory or diagnostic tests are not readily available in most areas, except for research purposes. PMID- 12721395 TI - Adverse reactions to aspirin and nonsteroidal antiinflammatory drugs (NSAIDs). AB - BACKGROUND: Aspirin can provoke reactions ranging from respiratory to cutaneous in those susceptible individuals. There has been particular attention looking at the role of cyclooxygenase enzymes and 2 and their role in aspirin exacerbated respiratory disease. OBJECTIVE: Patients who present with a spectrum of allergic and pseudoallergic reactions to aspirin pose a special challenge for the physician. This article discusses proposed classification system, clinical manifestations, pathogenesis of disease, and current treatment options of aspirin related disease. METHODS: Relevant articles in the medical literature were. derived from searching the MEDLINE database. Sources also include review articles, randomized control trials, and standard textbooks of allergy and immunology. RESULTS: Aspirin-exacerbated respiratory disease remains a complex, heterogeneous disease with manvaried clinical presentations. There have been many advances in trying to elucidate the pathogenesis of this disease. The classification system presented will provide greater ease when reading the literature and communicating with one another. Oral aspirin challenge remains the diagnostic test of choice for both respiratory and cutaneous reactions. Aspirin desensitization is an option for those with refractory respiratory disease or who require aspirin for other medical conditions. CONCLUSIONS: This review discusses the challenges in classification, diagnosis and treatment of those patients with a sensitivity to aspirin. Special attention is made to the possible mechanisms mediating disease progression and how specific. Therapies, such as the leukotriene modifiers may be helpful. PMID- 12721396 TI - Hypersensitivity reactions to chemotherapeutic drugs. AB - There is an ever-increasing number of therapeutics used to treat cancer. A recent publication listed 86 currently available antineoplastic medications. Despite this large number, hypersensitivity reactions are not common except with platinum compounds (cisplatin, carboplatin), epipodophyllotoxins (teniposide, etoposide), asparaginase, taxanes (paclitaxel), and procarbazine. Doxorubicin and 6 mercaptopurine are occasionally associated with hypersensitivity reaction. Comparable reactions with other chemotherapeutic agents are. uncommon; many are only anecdotal reports. Reactions associated with individual drugs are discussed in detail. The mechanisms responsible for most of these reactions are not known, as they have generally not been evaluated. The term "hypersensitivity" is widely used in the chemotherapy literature without a common definition. Hypersensitivity is defined here as an unexpected reaction with signs and symptoms not consistent with known toxicity of the drug. Most reactions are coincident with or within hours of drug administration. Almost all are associated with parenteral administration. Symptoms include flushing, alterations in heart rate and blood pressure, dyspnea and bronchospasm, back pain, fever, pruritus, nausea and all types of rashes. Some cases may be due to non-immune mediated release of histamine or cytokines, as many patients can subsequently tolerate re-exposure after pretreatment with steroids and antihistamine, and slow readministration of the drug. This is more compatible with a graded challenge, than desensitization and is generally successful for taxanes, less so for platinum compounds. In most cases hypersensitivity reactions are associated with the specific chemotherapeutic drug. Reaction rates may vary with different forms of the drugs, e.g. pegylated. Occasionally excipients such as Cremaphor EL may induce hypersensitivity reactions. PMID- 12721397 TI - Adverse reactions to vaccines. AB - (The opinions or assertions contained herein are the private views of the authors and are not to be construed as official or as reflecting the views of the Department of the Army or the Department of Defense.) Immunization healthcare is becoming increasingly complex as the number and types of vaccines have continued to expand. Like all prescription drugs, vaccines may be associated with adverse events. The majority of these reactions are self-limited and not associated with prolonged disability. The media, Internet and public advocacy groups have focused on potentially serious vaccine-associated adverse events with questions raised about causal linkages to increasing frequencies of diseases such as autism and asthma. Despite a lack of evidence of a causal relationship to a variety of vaccine safety concerns, including extensive reviews by the Institute of Medicine, questions regarding vaccine safety continue to threaten the success of immunization programs. Risk communication arid individual risk assessment is further challenged by the public health success of vaccine programs creating the perception that certain vaccines are no longer necessary or justified because of the rare reaction risk. There is a need for improved understanding of true vaccine contraindications and precautions as well as host factors and disease threat in order to develop a patient specific balanced risk communication intervention. When they occur, vaccine related adverse events must be treated, documented and reported through the VAERS system. The increasing complexity of vaccination health care has led the Center of Disease Control and Prevention (CDC) to identify Vaccine Safety Assessment and Evaluation as a potential new specialty. PMID- 12721399 TI - Selenium, copper, and zinc concentrations in intestinal cancer tissue and in colon and rectum polyps. AB - The prospective randomized trial was used to determine Se, Zn, and Cu concentrations in intestinal cancer tissue and colorectal polyp. We also determined the relationship among the trace element levels in cancer tissue, the localization of neoplasms, and the stage of their development. The concentrations of these trace elements were examined in cancer tissue of the colorectum in 67 patients and in the colon and rectum polyps in 42 patients using the total reflection X-ray fluorescence (TRXRF) method. The mean concentration of Se in colorectal cancer was 0.86 microg/g tissue and was statistically higher than in the case of polyps (0.57 microg/g). The mean concentration of Zn in colorectal cancer was higher than in the polyp (14.8 microg/g and 9.84 microg/g, respectively). The determined average concentration of Cu in colorectal cancer was 3.87 microg/g tissue and was a little lower than the level of this metal in the polyp (3.94 microg/g). There was no difference in the levels of these trace elements depending on the location of the neoplasm and the stage of its development. Also, there was no difference between the concentrations of these trace elements in the cancer tissue of malignant and benign tumors after taking into consideration the sex and age of patients. During the examination, we determined significantly higher concentrations of only selenium and zinc in the cancer tissue and not in the polyp. The level of copper was lower in a malignant tumor than in a benign one. PMID- 12721400 TI - Changes of total content of magnesium and zinc status in patients with chronic toxoplasmosis. AB - The aim of this study was to investigate the effects of the Toxoplasma gondii infection on the total content of the essential elements Zn and Mg levels. Plasma zinc and magnesium concentrations were measured in 158 patients whose anti-T gondii IgG antibodies were positive. Scores were obtained for the patients and their age- and sex-matched 82 sero-negative healthy controls. The mean concentration of Mg in blood was significantly lower in sero-positive patients than in their controls both in females (p < 0.01) and males (p < 0.01). The average Zn concentration in seropositive female patients was 0.72+/-0.2 mg/L and 0.65+/-0.1 mg/L in controls (p > 0.05). The mean values of the Zn in blood were 1.00+/-0.3 mg/L in seropositive male patients and 0.84+/-0.2 in controls (p > 0.05). No correlation could be demonstrated between age and mean values of Zn and Mg in both sero-positive females/males and controls (p > 0.05). No significant correlation could be found between blood Zn and Mg levels in seropositive female/male patients and controls (p > 0.05). Mg levels were found to be clearly decreased in the patients with chronic toxoplasmosis, whereas no change was observed in Zn levels. PMID- 12721401 TI - Arsenic content in the femur head of the residents of southern and central Poland. AB - Arsenic content was assayed in the samples of the femur head of the people living in southern and central Poland (Krakow, n=13; Silesian region, n=13; Lodz, n=12). The average age being 68.7+/-8.7 yr. Arsenic content in the femur head was determined applying the hydride generation atomic absorption spectrometry (HG AAS) method after microwave mineralization. The average arsenic contents in the femur head of the residents of the Lodz, Krakow, and Silesian regions were 0.41 microg/g, 0.37 microg/g, and 0.18 microg/g, respectively. No correlation has been found between arsenic content in the femur head and the content of other metals. Neither the age nor sex of the people tested affected the arsenic content in the femur head. PMID- 12721402 TI - Comparison of whole-blood glutathione peroxidase activity, levels of serum selenium, and lipid peroxidation in subjects from the fishing and rural communities of "Rabo de Peixe" village, San Miguel Island, the Azores' Archipelago, Portugal. AB - The activity of glutathione peroxidase (GSH-Px), serum selenium (Se), and thiobarbituric acid reactive substances (TBARS) were measured in the whole blood of 148 healthy adults aged 20-60 yr from the fishing and rural communities of "Rabo de Peixe," The Azores, Portugal. The subjects did not live in the same household and had different socioeconomic profiles and dietary habits. The serum lipid profile and selected life habits were also considered in this study. No significant differences in the activity of GSH-Px were found in the interpopulation or intrapopulation analyses, classified by age or lipid profile. An age-dependent GSH-Px increase was noted in the younger male (M) subgroups (20 39 yr). The Se levels were higher in fishers (f) of both genders (M, F) than in subjects living in the rural (r) environment: 110+/-25 microg/L (f, M), 89+/-20 microg/L (f, F), 88+/-22 microg/L (r, M) and 80+/-17 microg/L (r, F). In the fishers, but not in the rural population, Se was higher in the males, but it did not show significant variation with age. The levels of TBARS were lower in the f than in the r male group. The Se level was lower and TBARS higher in the hyperlipemic women in the f group, compared to the corresponding controls. Our results suggest that the fishers (mainly men) show a better antioxidant status than that of their rural counterparts, due to differences in dietary habits between the study populations and between genders. PMID- 12721403 TI - Zinc inhibition of hepatic fructose metabolism in rats. AB - The ability of Zn to modulate key metabolic processes was investigated in a study of gluconeogenesis in isolated hepatocytes from fasted rats. Zn (100 microM) inhibited glucose production from fructose by 41%, sorbitol by 28%; glycerol by 17%, and glyceraldehyde by 26%. Maximum inhibition of gluconeogenesis from fructose occurred at 25 microM Zn. Zn inhibited the rate of lactate production from fructose by 24% but not from sorbitol, glycerol, or glyceraldehyde. Fructose uptake by hepatocytes was not affected by Zn. A positive linear relationship (r=0.994) was obtained between inhibition by Zn of glucose and lactate production, indicating that a common step in both pathways is inhibited by Zn. The effect of Zn on fructokinase, aldolase-B, and triokinase activities was determined on semipurified rat liver enzyme preparations. Zn had no affect on triokinase activity but inhibited the two other enzymes in a dose-dependent manner, with the inhibition of aldolase-B being much greater than of fructokinase for concentrations of Zn between 2.5 and 20 microM. Zn increased the intracellular concentration of fructose-1-P in hepatocytes incubated with fructose, indicating a more potent Zn inhibition of aldolase-B than fructokinase. In addition, hepatocytes treated with Zn had decreased ATP and ADP concentrations, but had normal energy charge, suggesting an effect of Zn on adenine nucleotide degradation or synthesis. The demonstration that Zn inhibits two enzymes in fructose metabolism adds to the growing list of metabolic pathways that are catalyzed by enzymes that are sensitive to Zn. PMID- 12721404 TI - Studies on the effects of x-ray on erythrocyte zinc and copper concentrations in rabbits after treatment with antioxidants. AB - The aim of this study was to investigate the effect of supplemental antioxidant vitamins and minerals on the erythrocyte concentrations of zinc and copper in rabbits after exposure to X-rays. The animals were divided into two experimental and one control group (CG). The first group (VG) was given daily oral doses of vitamins E and C; supplemental amounts of manganese, zinc, and copper were mixed with the feed and given to the second group of experimental animals (MG). Blood samples were taken from all groups before and after 4 wk of vitamin and mineral administration and after irradiation with a total dose of 550-rad X-rays. The administration of minerals caused the most significant increases of Zn and Cu. Even after irradiation, the zinc levels in the irradiated animals were higher than in the nonirradiated vitamin-supplemented animals (p<0.05). The results suggest that supplementation with antioxidant vitamins and minerals may have a protective effect against X-ray-induced damage. PMID- 12721405 TI - Study of the thermokinetic properties of copper(II) on Escherichia coli growth. AB - By using an LKB2277 Bioactivity Monitor, stop-flow mode, the power-time curves of Escherichia coli at 37 degrees C affected by Cu(II) were determined. Some parameters, such as growth rate constants k, inhibitory ratio I, the heat output Qlog in the log phase, and the generation times G were obtained. According to these parameters, we found that a low concentration of Cu(II) (0-20 microg/mL) had an promoting action on the growth of E. coli, but a high concentration of Cu(II) (40-100 microg/mL) had an inhibitory action. The toxicity of Cu(II) can also be expressed as the half-inhibitory concentration IC50; the value is 69.7 microg/mL. The assay is quantitative, inexpensive, and versatile. PMID- 12721406 TI - Microcalorimetric investigation of the effect of manganese(II) on the growth of Tetrahymena shanghaiensis S199. AB - The heat output of and the effect of manganese (II) on Tetrahymena shanghaiensis S199 growth metabolism has been determined by means of a LKB-2277 BioActivity monitor. Different concentrations of manganese(II) ions have different effects on the growth of T. shanghaiensis. At low concentrations (0-40 microg/mL) culture growth is promoted, whereas high concentrations (60-800 microg/mL) slow growth. Furthermore, concentrations of 1200 microg/mL or greater stop the growth of this protozooa completely. PMID- 12721407 TI - Microcalorimetric studies of the action of Na2SeO3 on the growth of Halobacterium halobium R1. AB - The effect of Na2SeO3 on the growth of Halobacterium halobium R1 was investigated by means of microcalorimetry at 37 degrees C. The biological response to toxicants is observed as the inhibition of the rate constant of growth of living cells. A low concentration of Na2SeO3 stimulated the growth of H. halobium R1, and a high concentration of Na2SeO3 inhibited the growth of H. halobium R1. Toxicity may be expressed as the half-inhibition concentration (IC50). The rate constants of growth (k) and the concentrations of Na2SeO3 (c) shows a linear relationship: k = 1.790 x 10(-6) -- 2.27 x 10(-3) c. The value of IC50 obtained from the accompanying figure of I-c is 679 microg/mL. PMID- 12721408 TI - Adsorption of components of enzymatic synthesis of ampicillin on different hydrophobic resins. AB - This work compared the performance of three hydrophobic resins for the adsorption of ampicillin (AMP), D-phenylglycine (PG), D-phenylglycine methyl ester (PGME), and 6- aminopenicillanic acid (6-APA). The influence of pH on adsorption efficiencies was assessed in the range of 4.5-8.5, at 4 and 25 degrees C. The values at 4 degrees C were slightly higher than those at 25 degrees C. The adsorption efficiency of AMP and 6-APA decreased at higher pHs, for the three resins. An opposite behavior was found for PGME, and the pH did not affect PG adsorption efficiency. Isotherm models were fitted to experimental equilibrium data and the best models were discriminated. PMID- 12721409 TI - Xylanase production by Trichoderma reesei rut C-30 on rice straw. AB - Xylanase production of Trichoderma reesei Rut C-30 was examined at different initial pH values (4.8, 5.9, and 7.0) on rice straw in shake flasks, and in a fermentor, for the best pH condition. Enzyme performance was tested on ammonia treated dwarf elephant grass. The maximum xylanase activities, 92 and 122 IU/mL, were obtained at pH 4.8 in the shake flasks and fermentor, respectively, in which good growth of the fungus was observed during the first 24 h and consumption of proteins dissolved from the rice straw caused the pH to rise later to values between 6.4 and 6.7 (optimal for xylanase production). The xylanases from T. reesei were as effective as Multifect XL, a commercial enzyme preparation, in hydrolyzing ammonia-treated elephant grass. PMID- 12721410 TI - A different method of measuring and detecting monoand dioxygenase activities: key enzymes in hydrocarbon biodegradation. AB - A spectrophotometric method of measuring oxygenase activity in cell extracts or in zymograms was developed. It is an easy and cheap method that allows spectrophotometric measurement of activity by a colored reaction and reveals activity bands in a polyacrylamide gel electrophoresis (PAGE) gel as brown bands. To prove its usefulness, we report on a study with the oxygenase present in strain YR-1, isolated from petroleum-contaminated soils, that uses hydrocarbons as its sole carbon source. Soluble oxygenase activity was detected (under our conditions of cellular homogenization) in the mycelium of a filamentous fungus strain named YR-1. Oxygenase activity from aerobically grown mycelium was detected in growth medium containing the hydrocarbons decane or hexadecane; the enzyme activity exhibited similar optimum pH for the hydroxylation of different aliphatic or aromatic substrates (decane, hexadecane, benzene, and naphthalene) to the corresponding alcohols. Zymogram analysis conducted with partially purified fractions from cell extracts from the aerobic mycelium of the YR-1 strain indicated the existence of only one oxygenase enzyme. Partially purified samples of enzyme, analyzed by sodium dodecyl sulfate PAGE, indicated the presence of one major protein band with a mol wt of 56 kDa that can be a constituent of the native enzyme. In samples of the enzyme, the 56-kDa protein gave a positive reaction in immunodetection experiments with antibodies directed against oxygenase from soybean. The partially purified enzyme oxidized different substrates, although higher activity was displayed with benzene. Km values obtained for benzene and decane indicated a higher affinity for the latter. PMID- 12721411 TI - Xylanase production by Penicillium canescens 10-10c in solid-state fermentation. AB - Filamentous fungi have been widely used to produce hydrolytic enzymes for industrial applications, including xylanases, whose levels in fungi are generally much higher than those in yeast and bacteria. We evaluated the influence of carbon sources, nitrogen sources, and moisture content on xylanase production by Penicillium canescens 10-10c in solid-state fermentation. Among agricultural wastes tested (wheat bran, untreated wheat straw, treated wheat straw, beet pulp, and soja meal), untreated wheat straw gave the highest production of xylanase. Optimal initial moisture content for xylanase production was 83%. The addition of 0.4 g of xylan or easily metabolizable sugar, such as glucose and xylose, at a concentration of 2 % to wheat straw enhanced xylanase production. In solid-state fermentation, even at high concentrations of glucose or xylose (10%), catabolic repression was minimized compared to the effect observed in liquid culture. Yeast extract was the best nitrogen source among the nitrogen sources investigated: peptone, ammonium nitrate, sodium nitrate, ammonium chloride, and ammonium sulfate. A combination of yeast extract and peptone as nitrogen sources led to the best xylanase production. PMID- 12721412 TI - Protease production by Streptomyces sp. isolated from Brazilian Cerrado soil: optimization of culture medium employing statistical experimental design. AB - Streptomyces are important microorganisms because of their capacity to produce numerous bioactive molecules. In the present work protease production, by Streptomyces sp. 594 isolated from a Brazilian Cerrado soil, was maximized by optimizing a low-cost culture medium composition (casitone and sugarcane molasses) using statistical experimental design. The final protease activity (56 U/mL) was 2.8-fold and 58-fold higher than that obtained in the beginning of this study, and in a previous work, using an actinomycete selection medium, respectively. Protease production, not growth associated, appeared to be modulated by an inducer system, whereby the C/N ratio seemed to play a significant role. PMID- 12721413 TI - Synthesis of monocaprin catalyzed by lipase. AB - The production of monoglyceride emulsifiers commonly employed in the food, cosmetic, and pharmaceutical industries can be catalyzed by lipases, biocatalysts that are becoming increasingly attractive in the enzyme market. The aim of this study was to produce monocaprin utilizing a commercial immobilized lipase (Lipozyme IM 20) through the direct esterification of capric acid and glycerol. Experiments were performed for 6 h in an open reactor and the products were analyzed by gas chromatography. The parameters investigated were the amount of enzyme, temperature, and molar ratio between the reagents (capric acid/glycerol). The experimental runs followed an experimental design generated using Statistica software. The results showed that all the parameters were significant and that monocaprin production was enhanced at the lower ranges of the tested variables. The best conditions established were 55 degrees C, 3% (w/w) enzyme concentration, and molar ratio of 1. The final product, obtained after 6 h of reaction, was 61.3% monocaprin, 19.9% dicaprin, and 18.8% capric acid. This composition satisfies the directives of the World Health Organization food emulsifiers, which requires that these mixtures have at least 70% mono- plus diglyceride, and a minimum of 30% monoacylglycerol. PMID- 12721414 TI - Effect of dose of xylanase on bleachability of sugarcane bagasse ethanol/water pulps. AB - Pulps obtained from the ethanol/water cooking of sugarcane bagasse were bleached with the xylanase enzyme obtained from the fungus Thermomyces lanuginosus IOC 4145 and with the commercial enzyme Cartazyme HS from Sandoz. By changing the enzyme dose from 4.3 to 36 IU/g of pulp, kappa number and viscosity were maintained when the xylanase from T. lanuginosus was used. On the other hand, by using Cartazyme HS, kappa number decreased by 17%, reaching 35.5. This pulp was further extracted with NaOH without a decrease in viscosity (10 cP), and pulp with a kappa number of 13 was obtained. Xylanases had no significant effect on the ethanol/water pulps. PMID- 12721415 TI - CelF of Orpinomyces PC-2 has an intron and encodes a cellulase (CelF) containing a carbohydrate-binding module. AB - A cDNA, designated celF, encoding a cellulase (CelF) was isolated from the anaerobic fungus Orpinomyces PC-2. The open reading frame contains regions coding for a signal peptide, a carbohydrate-binding module (CBM), a linker, and a catalytic domain. The catalytic domain was homologous to those of CelA and CelC of the same fungus and to that of the Neocallimastix patriciarum CELA, but CelF lacks a docking domain, characteristic for enzymes of cellulosomes. It was also homologous to the cellobiohydrolase IIs and endoglucanases of aerobic organisms. The gene has a 111-bp intron, located within the CBM-coding region. Some biochemical properties of the purified recombinant enzyme are described. PMID- 12721417 TI - Effect of aeration on lignin peroxidase production by Streptomyces viridosporus T7A. AB - The effect of aeration on lignin peroxidase production by Streptomyces viridosporus T7A was studied in a bench-scale bioreactor using a previously optimized growth medium (0.65% yeast extract and 0.1% corn oil, pH 7.0) at 37 degrees C and natural pH. Airflow rates of 0.3, 1.0, and 1.5 vvm and a fixed agitation of 200 rpm were initially studied followed by 1.0 vvm and 200, 300, 400, and 500 rpm. The use of 1.0 vvm and 400 rpm increased enzyme concentration 1.8-fold (100-180 U/L) and process productivity 4.8-fold (1.4-6.7 U/[L h]) in comparison with the use of 200 rpm and 0.3 vvm. The inexpensive corn oil, used as carbon source, besides its antifoam properties, proved to be nonrepressive for enzyme production. PMID- 12721416 TI - Partition behavior and partial purification of hexokinase in aqueous two-phase polyethylene glycol/citrate systems. AB - This study dealt with the partition behavior and partial purification of hexokinase (HK) from baker's yeast by liquid-liquid extraction using aqueous two phase polyethylene glycol (PEG)/citrate systems. First, we investigated the effect of agitation type (vortex and 8 rpm rotation) on the stability of the system, and then the effects of sodium citrate concentration, PEG concentration, and molar mass of PEG on the partition coefficient of this enzyme by using a 25 factorial experimental design. The results of this factorial experiment showed the possibility of a partial purification of HK by using two extraction steps, since the enzyme preferentially migrated to the top phase and the total proteins (mainly contaminants) remained in the bottom phase. The purification factor (PurTOP) of the enzyme in the top phase was 1.87, and the partition coefficient of the total proteins (KProt) was 0.47. PMID- 12721418 TI - Evaluation of supports and methods for immobilization of enzyme cyclodextringlycosyltransferase. AB - An experimental design with factorial planning was used for the immobilization of the enzyme cyclodextringlycosyltransferase (CGTase) from Bacillus firmus (strain no. 37) to select the best combination of support, method of immobilization, and conditions that gives primarily higher average values for the specific immobilized enzyme activity, and secondarily, higher average values for the percentage of protein fixation. The experimental design factors were as follows: supports-controlled-pore silica, chitosan, and alumina; immobilization methods adsorption, and two covalent bonding methods, either with gamma aminopropyltriethoxysilane or hexamethylenediamine (HEMDA); conditions-7 degrees C without agitation and 26 degrees C with stirring. The best combination of factors that lead to higher average values of the response variables was obtained with immobilization of CGTase in silica with HEMDA at 7 degrees C. However, immobilization in chitosan at 7 degrees C gave the highest immobilized CGTase specific activity, 0.25 micromole of beta-CD/ (min mg protein). Physical adsorption gave low specific enzyme activities, and, in general, a high load of enzyme leads to lower specific enzyme activity. PMID- 12721419 TI - Active nuclear shuffling system using a swollen conidium of Trichoderma reesei. AB - Cellulase hyperproducers of Trichoderma reesei can be constructed using autopolyploidization and haploidization techniques. To increase the efficiency of this method, the active nuclear shuffling system in a swollen conidium was effective. A dried mature green conidium of a model strain, T. reesei QM6a (IFO 31326), was swollen to make room for a larger autopolyploid nucleus. After colchicine treatment, a larger autopolyploid nucleus was produced in such a swollen conidium. Benomyl treatment of swollen conidia generated multiple smaller nuclei from one larger autopolyploid nucleus. Those smaller nuclei were transported through conidia to mycelia after germination. This system could contribute to increasing the efficiency of genetic shuffling. PMID- 12721421 TI - Phenylboronate-chitosan resins for adsorption of beta-amylase from soybean extracts. AB - Isolation and purification of bioproducts from crude extracts can be obtained by affinity methods based on reversible binding of a specific molecule to ligand immobilized in a porous matrix. In the present work, nicrospheres based on chitosan matrix, which incorporated aminophenylboronic acid as a derivative, were prepared and characterized, aimed at developing a beta-amylase adsorption process. Kinetic curves and adsorption isotheriru of the crude extracts as well as the breakthrough curves for a frontal chromatographic separation method of a commercial sample of beta-amylase from soybean are presented. These results were compared to similar data obtained with a comercial microspheres gel based-on agarose. PMID- 12721422 TI - Ester fuels and chemicals from biomass. AB - Bench-scale research demonstrated that using an efficient esterification step to integrate an ethanol with a carboxylic acid fermentation stream offers potential for producing valuable ester feedstocks and fuels. Polar organic acids from bacterial fermentations are difficult to extract and purify, but formation of the ammonium salts and their conversion to esters facilitates the purifications. An improved esterification procedure gave high yields of esters, and this method will lower the cost of ester production. Fuel characteristics have been determined for a number of ester-gasoline blends with promising results for lowering Reid vapor pressure and raising octane numbers. PMID- 12721423 TI - Purification of glucose-6-phosphate dehydrogenase from baker's yeast in aqueous two-phase systems with free triazine dyes as affinity ligands. AB - To improve the selectivity of glucose-6-phosphate dehydrogenase (G6PDH) extraction by an aqueous two-phase system, a simple and inexpensive affinity aqueous two-phase system using unbound reactive triazine dyes as ligands was introduced. In a polyethylene glycol (PEG)/hydroxypropyl starch (PES) system, the unbound free triazine dyes, Cibacron Blue F3GA and Procion Red HE3B, partitioned unevenly in the top PEG-rich phase and thus showed an affinity effect on G6PDH, but no influence on hexokinase. The various parameters investigated were pH of the system, buffers, molecular weight of PEG, and ligand type and concentration. A two-step affinity extraction process was established for the purification of G6PDH from baker's yeast. The total yield of G6PDH was 66.9% and purification factor was 2.35. PMID- 12721424 TI - Continuous clavulanic acid adsorption process. AB - Adsorption kinetics and equilibrium data of clavulanic acid, a beta-lactam antibiotic, on ion-exchange resin Amberlite IRA 400 were utilized to carry out the modeling and simulation of a continuous adsorption process. These simulations allowed the estimation of yield, concentration, and purification factors of the process utilizing the product final concentration. Experimental runs of this process were carried out using the conditions pointed out by simulation studies. Comparison of the experimental results and those calculated by the proposed model showed that the model could describe very well the main features of the continuous process. PMID- 12721425 TI - Extraction of nutraceuticals from milk thistle: I. Hot water extraction. AB - Milk thistle contains compounds that display hepatoxic protection properties. We examined the batch extraction of silymarin compounds from milk thistle seed meal in 50, 70, 85, and 100 degrees C water as a function of time. After 210 min of extraction at 100 degrees C, the yield of taxifolin was 1.2 mg/g of seed, a 6.2 fold increase over the results obtained in a Soxhlet extraction with ethanol on pretreated (defatted) seeds. Similarly, the yield of silychristin was 5.0 mg/g of seed, a 3.8-fold increase. The yields of silybinin A and silybinin B were 1.8 and 3.3 mg/g of seed, respectively, or roughly 30% of the Soxhlet yield. The ratios of the extracted compounds, and particularly the ratios at long extraction times, showed that the more polar compounds (taxifolin and silychristin) were preferentially extracted at 85 degrees C, while the less polar silybinin was favored at 100 degrees C. PMID- 12721426 TI - Extraction of nutraceuticals from milk thistle: part II. Extraction with organic solvents. AB - Seeds from milk thistle (Silybum marianum Gaert L.) contain flavanolignan and dihydroflavanol compounds that have interesting and important therapeutic activities. The recovery of these silymarin compounds generally involves a two step defatting and extraction process using organic solvents. This study examined the batch, single-stage extraction of whole and defatted seeds using ethanol, methanol, acetonitrile, and acetone as the solvents. In extracting defatted milk thistle seeds with organic solvents, extraction with ethanol resulted in the highest silymarin yield, although some potential degradation was observed. The maximum yields of taxifolin, silychristin, silydianin, silybinin A, and silybinin B in ethanol were 0.6, 4.0, 0.4, 4.0, and 7.0 mg/g of defatted seed, respectively. However, if silybinin A were the diastereoisomer of choice, methanol would be the preferred extraction solvent because it yielded the highest silybinin A to silybinin B ratio. Interestingly, lipid removal is an important extraction step, because defatted material yields twice the silymarin concentration. PMID- 12721427 TI - Effect of lidocaine on ovalbumin and egg albumin foam stability. AB - Foam fractionation is a simple separation process that can remove and concentrate hydrophobic molecules such as proteins, surfactants, and organic wastes from an aqueous solution. Bovine serum albumin and ovalbumin have been widely used as model proteins due to their strong foaming potential and low price. Here, we study the effect of lidocaine on albumin foam, since drugs like lidocaine are known to bind with albumin. We observed that lidocaine not only enhances the amount of foam produced but also the stability of that foam as well. The foam stability was evaluated as the decay rate constant of the foam, determined from a change in height (or volume) of the foam over a given time period. PMID- 12721428 TI - Estimation of solubility effect on the herbicide controlled-release kinetics from lignin-based formulations. AB - Understanding the main phenomena involved in the controlled-release kinetics of herbicides in a water bath is a very important requisite for diffusive- parameter estimation, because, some mathematical models based on Fick's second law for diffusion have been developed to describe the controlled-release kinetic data. However, the validity of these models is restricted to the following assumptions: (1) the formulation is an isothermal slab; (2) the release occurs through the two faces of the slab; (3) the herbicide is dissolved in the water contained in the slab pores at a concentration less than the saturation concentration (cis); (4) the total sum of the individual volumes of the pores is epsilonAL (epsilon is the slab porosity, A is the slab area, and L is the slab thickness); and (5) the initial concentration of herbicide in the pores is M0/epsilonAL (M0 is the initial amount of herbicide in the matrix). The fourth assumption may be invalid for mathematical description of systems in which the herbicide concentration in the slab may be above the saturation concentration. If this were true, the final assumption would also be invalid, because the initial concentration of herbicide in the pores is cis in this case. This work presents a study of the solubility effect on the controlled-release kinetics of herbicides from lignin matrices. PMID- 12721429 TI - Permeation associated with three-phase-partitioning method on release of green fluorescent protein. AB - Transformed cells of Escherichia coli expressing recombinant green fluorescent protein (GFPuv) were subjected to two methods of extraction: (1) freezing/thawing/sonication (FTS) cycles prior to the three-phase partitioning (TPP) method, or (2) directly to TPP extraction. The amount of GFPuv released by the FTS plus TPP method varied: 374 microg/mL (first cycle), 93-442 microg/mL (second cycle), 32-359 microg/mL (third cycle), 18-115 microg/mL (fourth cycle). The GFPuv yields by the second method (TPP only) were, 23-54 microg/mL for the first extract and 33-91 microg/mL for the second. The FTS plus TPP method released similar amounts of GFPuv to that extracted by TPP; and provided a better mixture elution through the hydrophobic interaction column: 13-63 microg/mL for FTS plus TPP methods, and 2.5-13 microg/mL for TPP. The results showed that although selective permeation is a more laborious methodology, it was more efficient for obtaining of GFPuv in relation to the direct extraction of the cells for TPP. PMID- 12721430 TI - Comparison of growth characteristics of Panax ginseng hairy roots in various bioreactors. AB - This study investigated the effects of flask-to-liquid volume ratio on the growth of Panax ginseng hairy root, transformed by Agrobacterium rhizogenes, in flask cultures and compared the characteristics of various bioreactors for scale-up. The flask-to-liquid volume ratio was optimum at 1.5 mL of air/mL of medium in flask cultures, and hairy root growth was not affected above the optimum ratio. In 500-mL flask culture, hairy root showed two growth phases. After the first exponential growth, specific growth rate decreased. The growth characteristics of P. ginseng hairy root in various bioreactors were investigated. Hairy root growth was about 55- fold of inoculum after 39 d in a 5-L bioreactor and about 38-fold of inoculum after 40 d in a 19-L bioreactor. Carbon yield was higher in a 19-L bioreactor than in others, but it did not show any linear relationship to the growth rate of hairy roots in bioreactors. PMID- 12721431 TI - Heterogeneous aspects of acid hydrolysis of alpha-cellulose. AB - Hydrolysis of alpha-cellulose by H2SO4 is a heterogeneous reaction. As such the reaction is influenced by physical factors. The hydrolysis reaction is therefore controlled not only by the reaction conditions (acid concentration and temperature) but also by the physical state of the cellulose. As evidence of this, the reaction rates measured at the high-temperature region (above 200 C) exhibited a sudden change in apparent activation energy at a certain temperature, deviating from Arrhenius law. Furthermore, alpha-cellulose, once it was dissolved into concentrated H2SO4 and reprecipitated, showed a reaction rate two orders of magnitude higher than that of untreated cellulose, about the same magnitude as cornstarch. The alpha-cellulose when treated with a varying level of H2SO4 underwent an abrupt change in physical structure (fibrous form to gelatinous form) at about 65% H2SO4. The sudden shift of physical structure and reaction pattern in response to acid concentration and temperature indicates that the main factor causing the change in cellulose structure is disruption of hydrogen bonding. Finding effective means of disrupting hydrogen bonding before or during the hydrolysis reaction may lead to a novel biomass saccharification process. PMID- 12721432 TI - Characterization of molecular weight distribution of oligomers from autocatalyzed batch hydrolysis of xylan. AB - Oat spelt xylan was treated with water in a batch reactor at temperaturesof 180 and 200 degrees C. Ion-moderated partition (IMP) chromatography was thenapplied to separate oligomers in solution according to their molecular size. Calibration of the IMP measurements based on peak height was found toquantify dissolved monomer and oligomer yields well. Oligomer concentrationsin the liquid hydrolysate were also determined from the difference inxylose monomer concentrations measured by high-performance liquid chromatographybefore and after posthydrolysis of dissolved xylooligosaccharidesto xylose. Delayed formation and then rapid disappearance of oligomersfrom DP10 to DP2 was observed by IMP, and total oligomer yields measuredby IMP and posthydrolysis were very similar at these times. However, whileIMP detected virtually no oligomers initially, posthydrolysis measurementsgave significant amounts of soluble oligomers at these times, indicating thatoligomers with chain lengths >10 were in solution but not detectable by theIMP system used. PMID- 12721433 TI - Conversion of sugarcane bagasse to carboxylic acids using a mixed culture of mesophilic microorganisms. AB - Using the MixAlco process, biomass can be converted into carboxylic acids, which can be chemically converted into mixed alcohol fuels. This study focused on the use of countercurrent fermentation to anaerobically convert sugarcane bagasse and chicken manure to mixed carboxylic acids using a mixed culture of mesophilic microorganisms from terrestrial and marine sources. Bagasse was pretreated with lime to increase digestibility. The continuum particle distribution model (CPDM) simulated continuous fermentors based on data collected from batch experiments. This model saves considerable time in determining optimum operating conditions. For an 80% bagasse/20% chicken manure fermentation with terrestrial inoculum at a volatile solids loading rate (VSLR) of 7.36 g/(L of liquid d) and a liquid residence time (LRT) of 8.88 d, total carboxylic acid productivity, total acid selectivity, and yield were 2.49 g/(L of liquid d), 0.581 g of total acid/ g of VS digested, and 0.338 g of total acid/g of VS fed, respectively, at a concentration of 18.7 g of total acid/L. At the same VSLR and LRT, fermentation with marine inoculum gave higher total acid productivity, total acid selectivity, and yield than fermentation with terrestrial inoculum. For an 80% bagasse/20% chicken manure fermentation with marine inoculum at a VSLR of 3.83 g/(L of liquid d) and an LRT of 12.1 d, total carboxylic acid productivity, total acid selectivity, and yield were 1.38 g/(L of liquid d), 0.667 g of total acid/g of VS digested, and 0.359 g of total acid/g of VS fed, respectively, at a concentration of 16.2 g of total acid/L. PMID- 12721434 TI - Alternative approach for utilization of pentose stream from sugarcane bagasse by an induced flocculent Pichia stipitis. AB - A new approach for the utilization of hemicellulosic hydrolysate from sugarcane bagasse is described. This approach consists of using the hydrolysate to dilute the conventional feedstock (sugarcane juice) to the usual sugar concentration (150 g/L) employed for the industrial production of ethanol. The resulting sugar mixture was used as the substrate to evaluate the performance of a continuous reactor incorporating a cell recycle module, operated at several dilution rates. An induced flocculent pentose-fermenting yeast strain was used for this bioconversion. Under the conditions used, the reactor performance was satisfactory at substrate feed rates of 30 g/(L h) or less, corresponding to an ethanol productivity of about 11.0 g/(L h) and an overall sugar conversion >95%. These results show real advantages over the existing alternatives for a better exploitation of surplus bagasse to increase industrial alcohol production. PMID- 12721435 TI - Hydrogen production from paper sludge hydrolysate. AB - The main objective of this study was to develop a system for the production of "renewable" hydrogen. Paper sludge is a solid industrial waste yielding mainly cellulose, which can be used, after hydrolysis, as a feedstock in anaerobic fermentation by (hyper)thermophilic organisms, such as Thermotoga elfii and Caldicellulosiruptor saccharolyticus. Tests on different medium compositions showed that both bacteria were able to produce hydrogen from paper sludge hydrolysate, but the amount of produced hydrogen and the requirement for other components differed. Hydrogen production by T. elfii strongly depended on the presence of yeast extract and salts. By contrast, C. saccharolyticus was less dependent on medium components but seemed to be inhibited by a component present in the sludge hydrolysate. Utilization of xylose was preferred over glucose by C. saccharolyticus. PMID- 12721436 TI - Single-stage anaerobic codigestion for mixture wastes of simulated Korean food waste and waste activated sludge. AB - Korean food waste was treated with a single-stage anaerobic codigester (SSAD) using waste activated sludge (WAS) generated from a municipal wastewater treatment plant. The stability and performance of the system was analyzed. The C/N ratio was improved with increasing food waste fraction of feed mixture. The pH, alkalinity, and free ammonia nitrogen concentration were the parameters used to evaluate the digester's stability. The experimentally determined values of the parameters indicated that there were no methane inhibitions in the digester. Digester performance was determined by measuring the total chemical oxygen demand TCOD), volate solids (VS) removal, methane content in biogas, methane production rate (MPR), and specific methane productivity. Methane content in biogas and MPR were significantly dependent on hydraulic retention time (HRT) and ratio of food waste to WAS. The methane content in biogas decreased at shorter HRT or higher organic loading rate (OLR) with increased food waste fraction. Concerning the performance of the codigester, the optimum operating condition of the SSAD was found to be at an HRT of 10 d with a feed mixture ratio of 50% food waste and 50% WAS. A TCOD removal efficiency of 53.6% and a VS removal efficiency of 53.7% were obtained at an OLR of 5.96 kg of TCOD/(m3 d) and 3.14 kg of VS/(m3 d), respectively. A maximum MPR of 1.15 m3 CH4/(m3 d) and an SMP of 0.37 m3 CH4/kg of VSfeed were obtained at an HRT of 10 d with a methane content of 63%. PMID- 12721437 TI - Biosorption and desorption of copper (II) ions by Bacillus sp. AB - Batch biosorption experiments were conducted to investigate the removal of Cu2+ ions from aqueous solutions by a series of bacterial strains isolated from a local activated sludge process. The characteristics of 12 isolates were identified and examined for their ability to bind Cu2+ ions from aqueous solution. Among the isolates, two species exhibited biosorption capacity >40 mg of Cu/g of dry cell. Isotherms for the biosorption of copper on bacterial cells were developed and compared, and the equilibrium data fitted well to the Langmuir and Freundlich isotherm models. The biosorption of copper increased significantly with increasing pH from 2.0 to 6.0 regardless of the species. More than 90% of copper sorbed on the cells of Bacillus sp. could be recovered by washing with 0.1 M HNO3 for 5 min. The performance of two different desorption processes was also tested and compared. The results show that five biosorption and desorption cycles are a better operation process than five successive biosorptions followed by one desorption to remove and recover copper from aqueous solution. The biosorbent could be used for at least five biosorptions and desorption cycles without loss of copper removal capacity. It can be concluded that the activated sludge or sludge-isolated bacteria could be a potential biosorbent for copper removal. PMID- 12721438 TI - Flow field in a shrinking-bed reactor for pretreatment of cellulosic biomass. AB - A shrinking-bed reactor was designed by the National Renewable Energy Laboratory to maintain a constant bulk packing density of cellulosic biomass. The high solid to-liquid ratio in the pretreatment process allows a high sugar yield and avoids the need to flush large volumes of solution through the reactor. The shrinking bed reactor is a promising pretreatment reactor with the potential for scale-up for commercial applications. To scale up the shrinking-bed reactor, it is necessary to understand the flow pattern in the reactor. In this study, flow field is simulated with computational fluid dynamics using a porous medium model. Different discrete "snapshots" and multiple steady states are utilized. The bulk flow pattern, velocity distribution, and pressure drop are determined from the simulation and can be used to guide reactor design and scale-up. PMID- 12721439 TI - Lactic acid production through cell-recycle repeated-batch bioreactor. AB - The effect of various nitrogen sources on cell growth and lactic acid production was investigated. The most effective nitrogen source was yeast extract; more yeast extract gave higher cell growth and lactic acid productivity. Yeast extract dosage and cell growth were proportional up to a yeast extract concentration of 30 g/L, and lactic acid productivity was linearly correlated up to a yeast extract dosage of 25 g/L. However, increasing the yeast extract content raises the total production cost of lactic acid. Therefore, we attempted to find the optimum yeast extract dosage for a repeated-batch operation with cell recycling. The results show that when using Enterococcus faecalis RKY1 only 26% of the yeast extract dosage for a conventional batch fermentation was sufficient to produce the same amount of lactic acid, whereas the lactic acid concentration in the product stream (92-94 g/L) and lactic acid productivity (6.03-6.20 g/[L x h]) were similar to those of a batch operation. Furthermore, long-term stability was established. PMID- 12721440 TI - Limits for alkaline detoxification of dilute-acid lignocellulose hydrolysates. AB - In addition to fermentable sugars, dilute-acid hydrolysates of lignocellulose contain compounds that inhibit fermenting microorganisms, such as Saccharomyces cerevisiae. Previous results show that phenolic compounds and furan aldehydes, and to some extent aliphatic acids, act as inhibitors during fermentation of dilute-acid hydrolysates of spruce. Treatment of lignocellulose hydrolysates with alkali, usually in the form of overliming to pH 10.0, has been frequently employed as a detoxification method to improve fermentability. A spruce dilute acid hydrolysate was treated with NaOH in a factorial design experiment, in which the pH was varied between 9.0 and 12.0, the temperature between 5 and 80 degrees C, and the time between 1 and 7 h. Already at pH 9.0, >25% of the glucose was lost when the hydrolysate was treated at 80 degrees C for 1 h. Among the monosaccharides, xylose was degraded faster under alkaline conditions than the hexoses (glucose, mannose, and galactose), which, in turn, were degraded faster than arabinose. The results suggest that alkali treatment of hydrolysates can be performed at temperatures below 30 degrees C at any pH between 9.0 and 12.0 without problems with sugar degradation or formation of inhibiting aliphatic acids. Treatment with Ca(OH)2 instead of NaOH resulted in more substantial degradation of sugars. Under the harsher conditions of the factorial design experiment, the concentrations of furfural and 5-hydroxymethylfurfural decreased while the total phenolic content increased. The latter phenomenon was tentatively attributed to fragmentation of soluble aromatic oligomers in the hydrolysate. Separate phenolic compounds were affected in different ways by the alkaline conditions with some compounds showing an increase in concentration while others decreased. In conclusion, the conditions used for detoxification with alkali should be carefully controlled to optimize the positive effects and minimize the degradation of fermentable sugars. PMID- 12721443 TI - Production of lactic acid from food wastes. AB - Conversion of food wastes into lactic acid by simultaneous saccharification and fermentation (SSF) was investigated. The process involves saccharification of the starch component in food wastes by a commercial amylolytic enzyme preparation (a mixture of amyloglucosidase, alpha-amylase, and protease) and fermentation by Lactobacillus delbrueckii. The highest observed overall yield of lactic acid in the SSF was 91% of theoretical. Lactic acid concentration as high as 80 g/L was attainable in 48 h of the SSF. The optimum operating conditions for the maximum productivity were found to be 42 degrees C and pH 6.0. Without supplementation of nitrogen-containing nutrients, the lactic acid yield in the SSF decreased to 60%: 27 g/L of lactic acid from 60 g/L of food waste. The overall performance of the SSF, however, was not significantly affected by the elimination of mineral supplements. PMID- 12721444 TI - Microbiologic oxidation of isosafrole into piperonal. AB - The biotransformation of isosafrole by Cladosporium sphaerospermum yielded piperonal, which is a compound of great commercial importance in the flavor and fragrance industries. The experiments were performed in 500-mL conical flasks containing 100 mL of Czapek-modified medium in an orbital shaker with controlled agitation and temperature. Spores of C. sphaerospermum were used as inocula, and after 96 h of incubation the substrate was added to the culture. Samples of 2 mL were withdrawn at 24-h intervals and analyzed by gas chromatography, (GC) and/or GC/MS spectroscopy. PMID- 12721445 TI - Breathing air from protein foam. AB - Protein foams can be used to extinguish fires. If foams are to be used to extinguish fires where people are present, such as in high-rise buildings or ships, then a method for allowing people to breathe in a foam-filled environment is needed. It is proposed that the air, used to create the foam be used for breathing. A canister that will break incoming air-filled foam has been designed for attachment to a standard gas mask, in order to provide breathable air to a trapped person. Preliminary results for the modified mask indicate feasibility of breathing air from air-filled protein foam. PMID- 12721447 TI - Partial purification and kinetic characterization of acid phosphatase from garlic seedling. AB - The objective of this study was to obtain purer acid phosphatases than produced by prior art by operating under conditions that improve the final product. The study features are the use of a mild nonionic detergent, 40-80% saturation with (NH4)2SOm4, maintained at low temperature to remove impurity, and the use of chromatografic columns to concentrate the acid phosphatase and remove non-acid phosphatase proteins with lower or higher molecular weights. Acid phosphatase was isolated and purified from garlic seedlings by a streamline method without the use of proteolytic and lipolytic enzymes, butanol, or other organic solvents. Grown garlic seedlings of 10- 15 cm height were homogenized with 0.1 M acetate buffer containing 0.1 M NaCl and 0.1% Triton X-100. After homogenization, the supernatant was filtered with paper filters. Filtrated supernatant was cooled to 4 degrees C, followed by a threestep fractionation of the proteins with ammonium sulfate. The crude enzyme was isolated as a green precipitate that was dissolved in a small amount of 0.1 M acetate buffer containing 0.1 M NaCl and 0.1% Triton X 100. Garlic seedling acid phosphatase was purified with ion-exchange chromatography (DEAE cellulose). The column was equilibrated with 0.1 M acetate buffer. Acid phosphatase was purified 40-fold from the starting material. The specific activity of the pure enzyme was 168 U/mg. A variety of stability and activity profiles were determined for the purified garlic seedling acid phosphatase: optimum pH, optimum temperature, pH stability, temperature stability, thermal inactivation, substrate specificity, effect of enzyme concentration, effect of substrate concentration, activation energy, and effect of inhibitor and activator. The molecular mass of acid phosphatase was estimated to be 58 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The optimum pH was 5.7 and the optimum temperature was 50 degrees C. The enzyme was stable at pH 4.0-10.0 and 40-60 degrees C. Activation energy was between 10 and 20 kcal, and as Michaelis Menten coefficients, Vm values were 100 and 20 mM/s and Km values were 21.27 and 8.33 mM for paranitrophenylphosphate and paranitrophenyl, respectively. Studies of the effect of metal ions on enzyme activity showed both an activating and a deactivating effect. While Cu, Mo, and Mn showed strong inhibitory effects, Na, Ca, and K were the significant activators of acid phosphatase. PMID- 12721448 TI - Automated filter paper assay for determination of cellulase activity. AB - Recent developments in molecular breeding and directed evolution have promised great developments in industrial enzymes as demonstrated by exponential improvements in beta-lactamase and green fluorescent protein (GFP). Detection of and screening for improved enzymes are relatively easy if the target enzyme is expressible in a suitable high-throughput screening host and a clearly defined and usable screen or selection is available, as with GFP and beta-lactamase. Fungal cellulases, however, are difficult to measure and have limited expressibility in heterologous hosts. Furthermore, traditional cellulase assays are tedious and time-consuming. Multiple enzyme components, an insoluble substrate, and generally slow reaction rates have plagued cellulase researchers interested in creating cellulase mixtures with increased activities and/or enhanced biochemical properties. Although the International Union of Pure and Applied Chemists standard measure of cellulase activity, the filter paper assay (FPA), can be reproduced in most laboratories with some effort, this method has long been recognized for its complexity and susceptibility to operator error. Our current automated FPA method is based on a Cyberlabs C400 robotics deck equipped with customized incubation, reagent storage, and plate-reading capabilities that allow rapid evaluation of cellulases acting on cellulose and has a maximum throughput of 84 enzyme samples per day when performing the automated FPA. PMID- 12721449 TI - D-Xylose transport by Candida succiphila and Kluyveromyces marxianus. AB - The kinetics and regulation of D-xylose uptake were investigated in the efficient pentose fermentor Candida succiphila, and in Kluyveromyces marxianus, which assimilate but do not ferment pentose sugars. Active high affinity (Km approximately 3.8 mM; Vmax approximately 15 nmol/[mg min]) and putative facilitated diffusion low-affinity (Km approximately 140 mM; Vmax approximately 130 nmol/[mg min]) transport activities were found in C. succiphila grown, respectively, on xylose or glucose. K. marxianus showed facilitated diffusion low affinity (Km approximately 103 mM; Vmax approximately 190 nmol/[mg min]) transport activity when grown on xylose under microaerobic conditions, and both a low-affinity and an active high-affinity (Km approximately 0.2 mM; Vmax approximately 10 nmol/[mg min]) transport activity when grown on xylose under fully aerobic conditions. PMID- 12721450 TI - Molecular characterization of a gene for aldose reductase ( CbXYL1) from Candida boidinii and its expression in Saccharomyces cerevisiae. AB - Candida boidinii produces significant amounts of xylitol from xylose, and assays of crude homogenates for aldose (xylose) reductase (XYL1p) have been reported to show relatively high activity with NADH as a cofactor even though XYL1p purified from this yeast does not have such activity. A gene coding for XYL1p from C. boidinii (CbXYL1) was isolated by amplifying the central region using primers to conserved domains and by genome walking. CbXYL1 has an open reading frame of 966 bp encoding 321 amino acids. The C. boidinii XYL1p is highly similar to other known yeast aldose reductases and is most closely related to the NAD(P)H-linked XYL1p of Kluyveromyces lactis. Cell homogenates from C. boidinii and recombinant Saccharomyces cerevisiae were tested for XYL1p activity to confirm the previously reported high ratio of NADH:NADPH linked activity. C. boidinii grown under fully aerobic conditions showed an NADH:NADPH activity ratio of 0.76, which was similar to that observed with the XYL1p from Pichia stipitis XYL1, but which is much lower than what was previously reported. Cells grown under low aeration showed an NADH:NADPH activity ratio of 2.13. Recombinant S. cerevisiae expressing CbXYL1 showed only NADH-linked activity in cell homogenates. Southern hybridization did not reveal additional bands. These results imply that a second, unrelated gene for XYL1p is present in C. boidinii. PMID- 12721451 TI - Changing flux of xylose metabolites by altering expression of xylose reductase and xylitol dehydrogenase in recombinant Saccharomyces cerevisiae. AB - We changed the fluxes of xylose metabolites in recombinant Saccharomyces cerevisiae by manipulating expression of Pichia stipitis genes (XYL1 and XYL2) coding for xylose reductase (XR) and xylitol dehydrogenase (XDH), respectively. XYL1 copy number was kept constant by integrating it into the chromosome. Copy numbers of XYL2 were varied either by integrating XYL2 into the chromosome or by transforming cells with XYL2 in a multicopy vector. Genes in all three constructs were under control of the strong constitutive glyceraldehyde-3-phosphate dehydrogenase promoter. Enzymatic activity of XR and XDH in the recombinant strains increased with the copy number of XYL1 and XYL2. XR activity was not detected in the parent but was present at a nearly constant level in all of the transformants. XDH activity increased 12-fold when XYL2 was on a multicopy vector compared with when it was present in an integrated single copy. Product formation during xylose fermentation was affected by XDH activity and by aeration in recombinant S. cerevisiae. Higher XDH activity and more aeration resulted in less xylitol and more xylulose accumulation during xylose fermentation. Secretion of xylulose by strains with multicopy XYL2 and elevated XDH supports the hypothesis that D-xylulokinase limits metabolic flux in recombinant S. cerevisiae. PMID- 12721452 TI - Effect of media on spore yield and thermal resistance of Bacillus stearothermophilus. AB - The interference of eight components in the yield of sporulation and thermal resistance to moist heat (121 degrees C) of Bacillus stearothermophilus spores suspended in 0.02 M calcium acetate solution and inoculated on paper strips previously treated with calcium acetate/calcium hydroxide was studied. The spore yield of 1.0 x 10(8)mL was developed at 62 degrees C in 17 media containing different concentrations of D-glucose, sodium chloride, L-glutamic acid, yeast extract, peptone, manganese sulfate, potassium phosphate, and ammonium phosphate. The combined effects of yeast extract, peptone, and glucose contributed positively to the spore yield and to the stability of the thermal resistance of both spores in suspension and on strips. PMID- 12721454 TI - A new oxygen sensitivity and its potential application in photosynthetic H2 production. AB - We have discovered a new competitive pathway for O2 sensitivity in algal H2 production that is distinct from the O2 sensitivity of hydrogenase per se. This O2 sensitivity is apparently linked to the photosynthetic H2 production pathway that is coupled to proton translocation across the thylakoid membrane. Addition of the proton uncoupler carbonyl cyanide-p-trifluoromethoxy- phenylhydrazone eliminates this mode of O2 inhibition on H2 photoevolution. This newly discovered inhibition is most likely owing to background O2 that apparently serves as a terminal electron acceptor in competition with the H2 production pathway for photosynthetically generated electrons from water splitting. This O2-sensitive H2 production electron transport pathway was inhibited by 3[3,4-dichlorophenyl]1,1 dimethylurea. Our experiments demonstrated that this new pathway is more sensitive to O2 than the traditionally known O2 sensitivity of hydrogenase. This discovery provides new insight into the mechanism of O2 inactivation of hydrogenase and may contribute to the development of a more-efficient and robust system for photosynthetic H2 production. PMID- 12721453 TI - Cassava flour wastewater as a substrate for biosurfactant production. AB - Five cassava flour wastewater (manipueira) preparations were tested as culture media for biosurfactant production by a wild-type Bacillus sp. isolate. No-solids (F), no-solids diluted (F/2), natural (I), natural diluted (I/2), and decanted (IPS) were the tested manipueira media. The microorganism was able to grow and to produce biosurfactant on all manipueira preparations. The media whose solids were removed (F and F/2) showed better results than preparations with the presence of solids (I, I/2, and IPS). No-solids medium (F) showed a surface tension of 26,59 mN/m and reciprocal of critical micelle concentration of over 100 and was selected as a potential substrate for biosurfactant production. PMID- 12721456 TI - Optimization of SO2-catalyzed steam pretreatment of corn fiber for ethanol production. AB - A batch reactor was employed to steam explode corn fiber at various degrees of severity to evaluate the potential of using this feedstock as part of an enzymatically mediated cellulose-to-ethanol process. Severity was controlled by altering temperature (150-230 degrees C), residence time (1-9 min), and SO2 concentration (0-6% [w/w] dry matter). The effects of varying the different parameters were assessed by response surface modeling. The results indicated that maximum sugar yields (hemicellulose-derived water soluble, and cellulose-derived following enzymatic hydrolysis) were recovered from corn fiber pretreated at 190 degrees C for 5 minutes after exposure to 3% SO2. Sequential SO2-catalyzed steam explosion and enzymatic hydrolysis resulted in a conversion efficiency of 81% of the combined original hemicellulose and cellulose in the corn fiber to monomeric sugars. An additional posthydrolysis step performed on water soluble hemicellulose stream increased the concentration of sugars available for fermentation by 10%, resulting in the high conversion efficiency of 91%. Saccharomyces cerevisiae was able to ferment the resultant corn fiber hydrolysates, perhydrolysate, and liquid fraction from the posthydrolysis steps to 89, 94, and 85% of theoretical ethanol conversion, respectively. It was apparent that all of the parameters investigated during the steam explosion pretreatment had a significant effect on sugar recovery, inhibitory formation, enzymatic conversion efficiency, and fermentation capacity of the yeast. PMID- 12721457 TI - A comprehensive kinetic model for dilute-acid hydrolysis of cellulose. AB - Acid-catalyzed hydrolysis is controlled not only by temperature and acid concentration but also by the physical state of the cellulose. Under low temperature and acid condition the cellulose structure stays in stable crystalline form. Therefore, the prevailing reaction mode is endwise hydrolysis. Glucose then becomes the main sugar product. However, when temperature and/or acid concentration is raised to a certain level, the cellulose structure becomes unstable by breakage of hydrogen bonding, the primary force that holds the cellulose chains. Once the crystalline structure of the cellulose is disrupted, acid molecules can penetrate into the inner layers of the cellulose chains. In support of this hypothesis, we have experimentally verified that a substantial amount of oligomers is formed as reaction intermediates under extremely low-acid and high-temperature conditions. We also found that the breakage of hydrogen bonds occurs rather abruptly in response to temperature. One such condition is 210 degrees C, 0.07% H2SO4. Glucose, once it is formed in the hydrolysate, interacts with acid-soluble lignin, forming a lignin-carbohydrate complex. This occurs concurrently with other reactions involving glucose such as decomposition and reversion. On the basis of these findings, a comprehensive kinetic model is proposed. This model is in full compliance with our recent experimental data obtained under a broad range of reaction conditions. PMID- 12721458 TI - Effect of agitation on removal of acetic acid from pretreated hydrolysate by activated carbon. AB - The effect of agitation on the adsorption of acetic acid by activated carbon was tested utilizing an external mass transfer-diffusion model. Simulated pretreated biomass was contacted with activated carbon under prescribed conditions of temperature and agitation. Adsorption isotherm studies are presented as well as batch kinetic rate studies. Use of these data enabled the determination of isotherm constants, an external mass transfer coefficient, and an effective diffusivity for each agitation rate studied. The external film coefficient results ranged from 33.62 microm/s to a complete absence of external mass transfer resistance, and the diffusivity results ranged from 0.8625 to 10.70 microm(2)/s. The optimum combination of no external film resistance, and highest diffusivity, 10.70 microm(2)/s, occurred at 250 rpm and 25 degrees C. The results of these models and the experimental parameters suggested an efficacious method and conditions for the removal of this undesirable chemical. PMID- 12721459 TI - Enzymatic digestibility of used newspaper treated with aqueous ammonia-hydrogen peroxide solution. AB - Wastepaper constitutes approximately half of municipal solid waste, making it a potential source of bioenergy. Newspaper was pretreated with an ammonia-hydrogen peroxide (H2O2) mixture in a shaking bath from room temperature to 80 degrees C, and then its enzymatic digestibility was measured. A significant amount of ink was removed from the newspaper slurry by the reciprocating movement of the shaking bath. In addition, the ammonia-H2O2 significantly swelled the substrate, thereby greatly increasing its susceptibility to enzymatic digestion. After pretreating the newspaper with conditions of 40 degrees C, 3 h, 130 strokes/min, and 4 wt% ammonia-2 wt% H2O2, the enzymatic digestibility was almost 90% of theoretical, or about 25% higher than that of untreated substrate. Digestibility was also investigated as a function of ammonia concentration, H2O2 concentration, shaking speed, pretreatment temperature, and time. PMID- 12721460 TI - Continuous production of butanol from starch-based packing peanuts. AB - Acetone, butanol, ethanol (ABE, or solvents) were produced from starch-based packing peanuts in batch and continuous reactors. In a batch reactor, 18.9 g/L of total ABE was produced from 80 g/L packing peanuts in 110 h of fermentation. The initial and final starch concentrations were 69.6 and 11.1 g/L, respectively. In this fermentation, ABE yield and productivity of 0.32 and 0.17 g/(L h) were obtained, respectively. Compared to the batch fermentation, continuous fermentation of 40 g/L of starchbased packing peanuts in P2 medium resulted in a maximum solvent production of 8.4 g/L at a dilution rate of 0.033 h-1. This resulted in a productivity of 0.27 g/(L h). However, the reactor was not stable and fermentation deteriorated with time. Continuous fermentation of 35 g/L of starch solution resulted in a similar performance. These studies were performed in a vertical column reactor using Clostridium beijerinckii BA101 and P2 medium. It is anticipated that prolonged exposure of culture to acrylamide, which is formed during boiling/autoclaving of starch, affects the fermentation negatively. PMID- 12721462 TI - Effect of Bacillus circulans D1 thermostable xylanase on biobleaching of eucalyptus kraft pulp. AB - The alkalophilic Bacillus circulans D1 was isolated from decayed wood. It produced high levels of extracellular cellulase-free xylanase. The enzyme was thermally stable up to 60 degrees C, with an optimal hydrolysis temperature of 70 degrees C. It was stable over a wide pH range (5.5-10.5), with an optimum pH at 5.5 and 80% of its activity at pH 9.0. This cellulase-free xylanase preparation was used to biobleach kraft pulp. Enzymatic treatment of kraft pulp decreased chlorine dioxide use by 23 and 37% to obtain the same kappa number (kappa number) and brightness, respectively. Separation on Sephadex G-50 isolated three fractions with xylanase activity with distinct molecular weights. PMID- 12721461 TI - Measurement of rheological properties of corn stover suspensions. AB - Corn stover is currently being evaluated as a feedstock for ethanol production. The corn stover suspensions fed to reactors typically range between 10 and 40% solids. To simulate and design bioreactors for processing highly loaded corn stover suspensions, the rheologic properties of the suspension must be measured. In systems with suspended solids, rheologic measurements are difficult to perform owing to settling in the measurement devices. In this study, viscosities of corn stover suspensions were measured using a helical ribbon impeller viscometer. A calibration procedure is required for the impeller method in order to obtain the shear rate constant, k, which is dependent on the geometry of the measurement system. The corn stover suspensions are described using a power law flow model. PMID- 12721463 TI - Fungi allergens produced by solid-state fermentation process: optimization and allergen characterization. AB - Allergenic extracts were produced from Drechslera (Helminthosporium) monoceras biomass cultured by solid-state fermentation using wheat bran as the substrate. The main fermentation variables were selected by statistical design, and the optimized biomass yield (1.43 mg/[g of dry substrate d]) was obtained at pH 9.5 and 45.8% moisture. The allergenic extracts were produced from crude extract by protein precipitation and polyphenol removal. Proteins in the range of 16-160 kDa were identified in the extracts. Their reactions in patients were characterized by in vivo cutaneous tests (positive in 40% of the atopic patients) and by dot blotting assays. PMID- 12721464 TI - Hybrid model for an enzymatic reactor: hydrolysis of cheese whey proteins by alcalase immobilized in agarose gel particles. AB - Cheese whey proteolysis, carried out by immobilized enzymes, can either change or evidence functional properties of the produced peptides, increasing the potential applications of this byproduct of the dairy industry. Optimization and scale-up of the enzymatic reactor relies on its mathematical model-a set of mass balance equations, with reaction rates usually given by Michaelis-Menten-like kinetics; no information about the distribution of peptides' molecular sizes is supplied. In this article, a hybrid model of a batch enzymatic reactor is presented, consisting of differential mass balances coupled to a "neural-kinetic model," which provides the molecular weight distributions of the resulting peptides. PMID- 12721465 TI - Preliminary investigation of fungal bioprocessing of wheat straw for production of straw-thermoplastic composites. AB - Straw utilization for composites is limited by poor resin and polymer penetration, and excessive resin consumption owing to the straw cuticle, fines, and lignin-hemicellulose matrix. White-rot fungi degrade these components of straw and could, therefore, potentially be used to improve resin penetration and resin binding without the use of physical or chemical pretreatments. Although long treatment times and large footprints the limit use of fungal treatments on a large scale, distributed fungal pretreatments could alleviate land requirements. In this article, we present progress toward the development of a passive fungal straw upgrading system utilizing whiterot fungi. PMID- 12721466 TI - Simulation of aerated lagoon using artificial neural networks and multivariate regression techniques. AB - The aim of this study was to develop an empirical model that provides accurate predictions of the biochemical oxygen demand of the output stream from the aerated lagoon at International Paper of Brazil, one of the major pulp and paper plants in Brazil. Predictive models were calculated from functional link neural networks (FLNNs), multiple linear regression, principal components regression, and partial least-squares regression (PLSR). Improvement in FLNN modeling capability was observed when the data were preprocessed using the PLSR technique. PLSR also proved to be a powerful linear regression technique for this problem, which presents operational data limitations. PMID- 12721467 TI - Simplistic modeling approach to heterogeneous dilute-acid hydrolysis of cellulose microcrystallites. AB - The classic kinetic model for cellulose hydrolysis is often referred to as pseudo homogeneous, a term revealing the insight that the process is actually heterogeneous. During the past 10-15 yr, the shortcomings of this model have been demonstrated in various studies and the interest in the heterogeneous aspects has increased. The present work presents a simplistic model in which the intrinsic, heterogeneous hydrolysis and transport rates are coupled by the assumption of a constant glucosidic surface concentration. The mechanisms affecting these two rates are largely unknown, but the model serves as a guideline for further exploration of the process. PMID- 12721468 TI - Cellulosic fuel ethanol: alternative fermentation process designs with wild-type and recombinant Zymomonas mobilis. AB - Iogen (Canada) is a major manufacturer of industrial cellulase and hemicellulase enzymes for the textile, pulp and paper, and poultry feed industries. Iogen has recently constructed a 40 t/d biomass-to-ethanol demonstration plant adjacent to its enzyme production facility. The integration of enzyme and ethanol plants results in significant reduction in production costs and offers an alternative use for the sugars generated during biomass conversion. Iogen has partnered with the University of Toronto to test the fermentation performance characteristics of metabolically engineered Zymomonas mobilis created at the National Renewable Energy Laboratory. This study focused on strain AX101, a xylose- and arabinose fermenting stable genomic integrant that lacks the selection marker gene for antibiotic resistance. The "Iogen Process" for biomass depolymerization consists of a dilute-sulpfuric acid-catalyzed steam explosion, followed by enzymatic hydrolysis. This work examined two process design options for fermentation, first, continuous cofermentation of C5 and C6 sugars by Zm AX101, and second, separate continuous fermentations of prehydrolysate by Zm AX101 and cellulose hydrolysate by either wildtype Z. mobilis ZM4 or an industrial yeast commonly used in the production of fuel ethanol from corn. Iogen uses a proprietary process for conditioning the prehydrolysate to reduce the level of inhibitory acetic acid to at least 2.5 g/L. The pH was controlled at 5.5 and 5.0 for Zymomonas and yeast fermentations, respectively. Neither 2.5 g/L of acetic acid nor the presence of pentose sugars (C6:C5 = 2:1) appreciably affected the high performance glucose fermentation of wild-type Z. mobilis ZM4. By contrast, 2.5 g/L of acetic acid significantly reduced the rate of pentose fermentation by strain AX101. For single-stage continuous fermentation of pure sugar synthetic cellulose hydrolysate (60 g/L of glucose), wild-type Zymomonas exhibited a four fold higher volumetric productivity compared with industrial yeast. Low levels of acetic acid stimulated yeast ethanol productivity. The glucose-to-ethanol conversion efficiency for Zm and yeast was 96 and 84%, respectively. PMID- 12721469 TI - Effects of pressure pulsation on oxygen transfer rate measured by sulfite method. AB - Pressure pulsation (PP) was investigated for its effects on oxygen transfer rate (OTR) measured by sulfite oxidation. By manipulating airflow rate, 0.41- 1.2 vvm, and a control valve in a 4-L bioreactor, the frequency of PP was varied at different gas pressures3-15 psig. A mathematical model of OTR was built and compared to experimental data. OTR was also examined at constant gas pressure, 4.5-15.0 psig. The results indicate a good agreement between measurement and model prediction. OTR above 9 psig during PP showed significant enhancement at 25 degrees C. This proves that PP not only affects the elevation of DO level, but also increases the interfacial area and mass transfer coefficient. PMID- 12721471 TI - Rapid biomass analysis: new tools for compositional analysis of corn stover feedstocks and process intermediates from ethanol production. AB - New, rapid, and inexpensive methods that monitor the chemical composition of corn stover and corn stover-derived samples are a key element to enabling the commercialization of processes that convert stover to fuels and chemicals. These new techniques combine near infrared (NIR) spectroscopy and projection to latent structures (PLS) multivariate analysis to allow the compositional analysis of hundreds of samples in 1 d at a cost of about $10 each. The new NIR/PLS rapid analysis methods can also be used to support a variety of research projects that would have been too costly to pursue by traditional methods. PMID- 12721472 TI - Xylem-specific and tension stress-responsive expression of cellulose synthase genes from aspen trees. AB - Genetic improvement of cellulose biosynthesis in woody trees is one of the major goals of tree biotechnology research. Yet, progress in this field has been slow owing to (1) unavailability of key genes from tree genomes, (2) the inability to isolate active and intact cellulose synthase complexes and, (3) the limited understanding of the mechanistic processes involved in the wood cellulose development. Here I report on the recent advances in molecular genetics of cellulose synthases (CesA) from aspen trees. Two different types of cellulose synthases appear to be involved in cellulose deposition in primary and secondary walls in aspen xylem. The three distinct secondary CesAs from aspen- PtrCesA1, PtrCesA2, and PtrCesA3-appear to be aspen homologs of Arabidopsis secondary CesAs AtCesA8, AtCesA7, and AtCesA4, respectively, based on their high identity/similarity (>80%). These aspen CesA proteins share the transmembrane domain (TMD) structure that is typical of all known "true" CesA proteins: two TMDs toward the N-terminal and six TMDs toward the C-terminal. The putative catalytic domain is present between TMDs 2 and 3. All signature motifs of processive glycosyltransferases are also present in this catalytic domain. In a phylogenetic tree based on various predicted CesA proteins from Arabidopsis and aspen, aspen CesAs fall into families similar to those seen with Arabidopsis CesAs, suggesting their functional similarity. The coordinate expression of three aspen secondary CesAs in xylem and phloem fibers, along with their simultaneous tension stress-responsive upregulation, suggests that these three CesAs may play a pivotal role in biosynthesis of better-quality cellulose in secondary cell walls of plants. These results are likely to have a direct impact on genetic manipulation of trees in the future. PMID- 12721473 TI - Microbial pretreatment of biomass: potential for reducing severity of thermochemical biomass pretreatment. AB - Typical pretreatment requires high-energy (steam and electricity) and corrosion resistant, high-pressure reactors. A review of the literature suggests that fungal pretreatment could potentially lower the severity requirements of acid, temperature and time. These reductions in severity are also expected to result in less biomass degradation and consequently lower inhibitor concentrations compared to conventional thermochemical pretreatment. Furthermore, potential advantages of fungal pretreatment of agricultural residues, such as corn stover, are suggested by its effectiveness in improving the cellulose digestibility of many types of forage fiber and agricultural wastes. Our preliminary tests show a three- to five fold improvement in enzymatic cellulose digestibility of corn stover after pretreatment with Cyathus stercoreus; and a ten- to 100-fold reduction in shear force needed to obtain the same shear rate of 3.2 to 7 rev/s, respectively, after pretreatment with Phanerochaete chrysosporium. PMID- 12721474 TI - Physical separation of straw stem components to reduce silica. AB - In this paper, we describe ongoing efforts to solve challenges to using straw for bioenergy and bioproducts. Among these, silica in straw forms a low-melting eutectic with potassium, causing slag deposits, and chlorides cause corrosion beneath the deposits. Straw consists principally of stems, leaves, sheaths, nodes, awns, and chaff. Leaves and sheaths are higher in silica, while chaff, leaves, and nodes are the primary sources of fines. Our approach to reducing silica is to selectively harvest the straw stems using an in-field physical separation, leaving the remaining components in the field to build soil organic matter and contribute soil nutrients. PMID- 12721475 TI - Application of a depolymerization model for predicting thermochemical hydrolysis of hemicellulose. AB - Literature data were collected and analyzed to guide selection of conditions for pretreatment by dilute acid and water-only hemicellulose hydrolysis, and the severity parameter was used to relate performance of different studies on a consistent basis and define attractive operating conditions. Experiments were then run to confirm performance with corn stover. Although substantially better hemicellulose sugar yields are observed when acid is added, costs would be reduced and processing operations simplified if less acid could be used while maintaining good yields, and understanding the relationship between operating conditions and yields would be invaluable to realizing this goal. However, existing models seldom include the oligomeric intermediates prevalent at lower acid levels, and the few studies that include such species do not account for the distribution of chain lengths during reaction. Therefore, the polymeric nature of hemicellulose was integrated into a kinetic model often used to describe the decomposition of synthetic polymers with the assumption that hemicellulose linkages are randomly broken during hydrolysis. Predictions of monomer yields were generally consistent with our pretreatment data, data reported in the literature, and predictions of other models, but the model tended to overpredict oligomer yields. These differences need to be resolved by gathering additional data and improving the model. PMID- 12721476 TI - Dilute-sulfuric acid pretreatment of corn stover in pilot-scale reactor: investigation of yields, kinetics, and enzymatic digestibilities of solids. AB - Corn stover is a domestic feedstock that has potential to produce significant quantities of fuel ethanol and other bioenergy and biobased products. However, comprehensive yield and carbon mass balance information and validated kinetic models for dilute-sulfuric acid (H2SO4) pretreatment of corn stover have not been available. This has hindered the estimation of process economics and also limited the ability to perform technoeconomic modeling to guide research. To better characterize pretreatment and assess its kinetics, we pretreated corn stover in a continuous 1 t/d reactor. Corn stover was pretreated at 20% (w/w) solids concentration over a range of conditions encompassing residence times of 3-12 min, temperatures of 165- 195 degrees C, and H2SO4 concentrations of 0.5-1.4% (w/w). Xylan conversion yield and carbon mass balance data were collected at each run condition. Performance results were used to estimate kinetic model parameters assuming biphasic hemicellulose hydrolysis and a hydrolysis mechanism incorporating formation of intermediate xylo-oligomers. In addition, some of the pretreated solids were tested in a simultaneous saccharification and fermentation (SSF) process to measure the reactivity of their cellulose component to enzymatic digestion by cellulase enzymes. Monomeric xylose yields of 69-71% and total xylose yields (monomers and oligomers) of 70-77% were achieved with performance level depending on pretreatment severity. Cellulose conversion yields in SSF of 80-87% were obtained for some of the most digestible pretreated solids. PMID- 12721477 TI - Hydrothermal pretreatment conditions to enhance ethanol production from poplar biomass. AB - Pretreatment has been recognized as a key step in enzyme-based conversion processes of lignocellulose biomass to ethanol. The aim of this study is to evaluate two hydrothermal pretreatments (steam explosion and liquid hot water) to enhance ethanol production from poplar (Populus nigra) biomass by a simultaneous saccharification and fermentation (SSF) process. The composition of liquid and solid fractions obtained after pretreatment, enzymatic digestibility, and ethanol production of poplar biomass pretreated at different experimental conditions was analyzed. The best results were obtained in steam explosion pretreatment at 210 C and 4 min, taking into account cellulose recovery above 95%, enzymatic hydrolysis yield of about 60%, SSF yield of 60% of theoretical, and 41% xylose recovery in the liquid fraction. Large particles can be used for poplar biomass in both pretreatments, since no significant effect of particle size on enzymatic hydrolysis and SSF was obtained. PMID- 12721478 TI - Estimation of temperature transients for biomass pretreatment in tubular batch reactors and impact on xylan hydrolysis kinetics. AB - A combined heat transfer/kinetic model was developed to quantify temperature variations in small tubular batch reactors and estimate the effect of deviations from isothermal operation on the kinetics of biomass pretreatment. Assuming that heat transfer was dominated by conduction in the radial direction, a classic parabolic time-dependent partial differential equation was applied to describe the temperature in the system and dedimensionalized to provide a single solution for application to all situations. A dimensionless expression for the reaction kinetics for xylan hydrolysis was then developed, and a single parameter expressed as the dimensionless ratio of the first-order rate constant times the tube radius squared divided by the thermal diffusivity was found to control the reaction rate. Three different characterizations of the deviation between the concentration profile predicted for isothermal xylan hydrolysis and that based on the transient temperature were directly related to this dimensionless rate constant parameter for both catalyzed and uncatalyzed hydrolysis kinetics. These results were then used to project the relationship between deviations in yield from isothermal results and the tube radius and reaction time. PMID- 12721480 TI - Combined use of H2SO4 and SO2 impregnation for steam pretreatment of spruce in ethanol production. AB - Fuel ethanol can be produced from softwood through hydrolysis in an enzymatic process. Prior to enzymatic hydrolysis of the softwood, pretreatment is necessary. In this study, two-step steam pretreatment employing dilute H2SO4 impregnation in the first step and SO2 impregnation in the second step, to improve the overall sugar and ethanol yield, was investigated. The first pretreatment step was performed under conditions of low severity (180 degrees C, 10 min, 0.5% H2SO4) to optimize the amount of hydrolyzed hemicellulose. In the second step, the washed solid material from the first pretreatment step was impregnated with SO2 and pretreated under conditions of higher severity to make the cellulose more accessible to enzymatic attack, as well as to hydrolyze a portion of the cellulose. A wide range of conditions was used in the second step to determine the most favorable combination. The temperatures investigated were between 190 and 230 degrees C, the residence times were 2, 5, and 10 min; and the SO2 concentration was 3%. The effect of pretreatment was assessed by both enzymatic hydrolysis of the solids and by simultaneous saccharification and fermentation (SSF) of the whole slurry, after the second pretreatment step. For each set of pretreatment conditions, the liquid fraction was also fermented to determine any inhibitory effects. Ethanol yield using the SSF configuration reached 66% of the theoretical value for pretreatment conditions in the second step of 210 degrees C and 5 min. The sugar yield using the separate hydrolysis and fermentation configuration reached 71% for pretreatment conditions of 220 degrees C and 5 min. PMID- 12721479 TI - Effect of sulfuric and phosphoric acid pretreatments on enzymatic hydrolysis of corn stover. AB - The pretreatment of corn stover with H2SO4 and H3PO4 was investigated. Pretreatments were carried out from 30 to 120 min in a batch reactor at 121 degrees C, with acid concentrations ranging from 0 to 2% (w/v) at a solid concentration of 5% (w/v). Pretreated corn stover was washed with distilled water until the filtrate was adjusted to pH 7.0, followed by surfactant swelling of the cellulosic fraction in a 0-10% (w/v) solution of Tween-80 at room temperature for 12 h. The dilute acid treatment proved to be a very effective method in terms of hemicellulose recovery and cellulose digestibility. Hemicellulose recovery was 62 90%, and enzymatic digestibility of the cellulose that remained in the solid was >80% with 2% (w/v) acid. In all cases studied, the performance of H2SO4 pretreatment (hemicellulose recovery and cellulose digestibility) was significantly better than obtained with H3PO4. Enzymatic hydrolysis was more effective using surfactant than without it, producing 10-20% more sugar. Furthermore, digestibility was investigated as a function of hemicellulose removal. It was found that digestibility was more directly related to hemicellulose removal than to delignification. PMID- 12721481 TI - Effect of lignocellulosic degradation compounds from steam explosion pretreatment on ethanol fermentation by thermotolerant yeast Kluyveromyces marxianus. AB - The filtrate from steam-pretreated poplar was analyzed to identify degradation compounds. The effect of selected compounds on growth and ethanolic fermentation of the thermotolerant yeast strain Kluyveromyces marxianus CECT 10875 was tested. Several fermentations on glucose medium, containing individual inhibitory compounds found in the hydrolysate, were carried out. The degree of inhibition on yeast strain growth and ethanolic fermentation was determined. At concentrations found in the prehy-drolysate, none of the individual compounds significantly affected the fermentation. For all tested compounds, growth was inhibited to a lesser extent than ethanol production. Lower concentrations of catechol (0.96 g/L) and 4-hydroxybenzaldehyde (1.02 g/L) were required to produce the 50% reduction in cell mass in comparison to other tested compounds. PMID- 12721482 TI - Enzymatic hydrolysis of ammonia-treated rice straw. AB - Rice straw pretreated with liquid anhydrous ammonia was hydrolyzed with cellulase, cellobiase, and hemicellulase. Ammonia-processing conditions were 1.5 g of NH3/g of dry matter, 85 degrees C, and several sample moisture contents. There were four ammonia addition time (min)-processing time (min) combinations. Sugars produced were analyzed as reducing sugars (dinitrosalicylic acid method) and by high-performance liquid chromatography. Monomeric sugars increased from 11% in the nontreated rice straw to 61% of theoretical in treated rice straw (79.2% conversion as reducing sugars). Production of monosaccharides was greater at higher moisture content and was processing time dependent. Glucose was the monosaccharide produced in greater amounts, 56.0%, followed by xylose, arabinose, and fructose, with 35.8, 6.6, and 1.4%, respectively. PMID- 12721483 TI - Effects of temperature and moisture on dilute-acid steam explosion pretreatment of corn stover and cellulase enzyme digestibility. AB - Corn stover is emerging as a viable feedstock for producing bioethanol from renewable resources. Dilute-acid pretreatment of corn stover can solubilize a significant portion of the hemicellulosic component and enhance the enzymatic digestibility of the remaining cellulose for fermentation into ethanol. In this study, dilute H2SO4 pretreatment of corn stover was performed in a steam explosion reactor at 160 degrees C, 180 degrees C, and 190 degrees C, approx 1 wt % H2SO4, and 70-s to 840-s residence times. The combined severity (Log10 [Ro] - pH), an expression relating pH, temperature, and residence time of pretreatment, ranged from 1.8 to 2.4. Soluble xylose yields varied from 63 to 77% of theoretical from pretreatments of corn stover at 160 and 180 degrees C. However, yields >90% of theoretical were found with dilute-acid pretreatments at 190 degrees C. A narrower range of higher combined severities was required for pretreatment to obtain high soluble xylose yields when the moisture content of the acidimpregnated feedstock was increased from 55 to 63 wt%. Simultaneous saccharification and fermentation (SSF) of washed solids from corn stover pretreated at 190 degrees C, using an enzyme loading of 15 filter paper units (FPU)/ g of cellulose, gave ethanol yields in excess of 85%. Similar SSF ethanol yields were found using washed solid residues from 160 and 180 degrees C pretreatments at similar combined severities but required a higher enzyme loading of approx 25 FPU/g of cellulose. PMID- 12721484 TI - Sugar monomer and oligomer solubility: data and predictions for application to biomass hydrolysis. AB - Oligomer solubility could potentially play an important role in controlling the rates and yields in the thermochemical hydrolysis of hemicellulose as a pretreatment for subsequent enzymatic conversion of cellulose. However, limited data or models are available to describe the aqueous solubility of sugar monomers and oligomers. In this work, we measured the solubilities of sugars common to many biomass feedstocks in the temperature range of 25-30 degrees C. Then we reviewed solubility models for sugars from the open literature. Finally, we applied models to test their ability to describe this and other data reported in the literature. It was found that the solubility of sugar monomers was not well described by the ideal solubility law or other more complex models. However, with an empirical adjustment to the enthalpy of fusion, the ideal solubility law was able to approximately predict the solubility of cello-oligomers. Based on these results, solubilities for low molecular weight xylo-oligomers are predicted to investigate their possible importance in pretreatment and define further experimental measurements needed to improve our understanding of sugar and oligomer solubility. PMID- 12721485 TI - Influence of pressure in ethanol/water pulping of sugarcane bagasse. AB - The influence of the pressure in the ethanol/water pulping of sugarcane bagasse was studied using argon pressure varying from 0.5 to 1.5 MPa. The reaction volume and activation volume were studied. For the reaction volume, temperature and time were constant and pressure was varied, and for the activation volume, temperature was constant and pressure and time were varied. The degradation of cellulose was not promoted by the pressure with positive reaction volume (4100 cm(3)/mol). On the other hand, degradation of xylan (polyoses) and lignin was strongly favored by the pressure and reaction volume ranged from -1000 to -3000 cm(3)/mol. PMID- 12721486 TI - Post-harvest processing methods for reduction of silica and alkali metals in wheat straw. AB - Silica and alkali metals in wheat straw limit its use for bioenergy and gasification. Slag deposits occur via the eutectic melting of SiO2 with K2O, trapping chlorides at surfaces and causing corrosion. A minimum melting point of 950 degrees C is desirable, corresponding to an SiO2:K2O weight ratio of about 3:1. Mild chemical treatments were used to reduce Si, K, and Cl, while varying temperature, concentration, % solids, and time. Dilute acid was more effective at removing K and Cl, while dilute alkali was more effective for Si. Reduction of minerals in this manner may prove economical for increasing utilization of the straw for combustion or gasification. PMID- 12721487 TI - Composition and ethanol production potential of cotton gin residues. AB - Cotton gin residue (CGR) collected from five cotton gins was fractionated and characterized for summative composition. The major fractions of the CGR varied widely between cotton gins and consisted of clean lint (5-12%),hulls (16-48%), seeds (6-24%), motes (16-24%), and leaves (14-30%). The summative composition varied within and between cotton gins and consisted of ash (7.9-14.6%), acid insoluble material (18-26%), xylan (4-15%),and cellulose (20-38%). Overlimed steam-exploded cotton gin waste was readily fermented to ethanol by Escherichia coli KO11. Ethanol yields were feedstock and severity dependent and ranged from 58 to 92.5% of the theoretical yields. The highest ethanol yield was 191 L (50 gal)/t, and the lowest was 120 L (32 gal)/t. PMID- 12721488 TI - Wood-ethanol for climate change mitigation in Canada. AB - The impetus for this paper is Canada's commitment under the United Nations Framework Convention on Climate Change to reduce national greenhouse gas emissions as well as reducing our dependency on fossil fuels. Wood-based ethanol offers an excellent opportunity for greenhouse gas mitigation due to market potential, an ability to offset significant emissions from the transportation sector, a reduction of emissions from CO2-intensive waste-management systems, and carbon sequestration in afforested plantations. While there are technological and economic barriers to overcome, using wood-biomass as a source of ethanol can be an economically viable tool for reducing greenhouse gas levels in the atmosphere. This paper examines the costs and mitigation potential of the production of ethanol from biomass supplied from industrial wood waste as well as from trees harvested from afforested land. PMID- 12721490 TI - Saccharification of marine microalgae using marine bacteria for ethanol production. AB - The saccharification of marine microalgae using amylase from marine bacteria in saline conditions was investigated. An amylase-producing bacterium, Pseudoalterimonas undina NKMB 0074 was isolated and identified. The green microalga NKG 120701 was determined to have the highest concentration of intracellular carbohydrate and was found from our algal culture stocks. P. undina NKMB 0074 was inoculated into suspensions containing NKG 120701 cells and increasingly reduced suspended sugars with incubation time. Terrestrial amylase and glucoamylase were inactive in saline suspension. Therefore, marine amylase is necessary in saline conditions for successful saccharification of marine microalgae. PMID- 12721491 TI - Efficient display of two enzymes on filamentous phage using an improved signal sequence. AB - Directed protein-evolution strategies generally make use of a link between a protein and the encoding DNA. In phage-display technology, this link is provided by fusion of the protein with a coat protein that is incorporated into the phage particle containing the DNA. Optimization of this link can be achieved by adjusting the signal sequence of the fusion. In a previous study, directed evolution of signal sequences for optimal display of the Taq DNA polymerase I Stoffel fragment on phage yielded signal peptides with a 50- fold higher incorporation of fusion proteins in phage particles. In this article, we show that for one of the selected signal sequences, improved display on phage can be generalized to other proteins, such as adenylate cyclases from Escherichia coli and Bordetella pertussis, and that this is highly dependent on short sequences at the C-terminus of the signal peptide. Further, the display of two enzymes on phage has been achieved and may provide a strategy for directing coevolution of the two proteins. These findings should be useful for display of large and cytoplasmic proteins on filamentous phage. PMID- 12721492 TI - Specific mutations within the alpha4-alpha5 loop of the Bacillus thuringiensis Cry4B toxin reveal a crucial role for Asn-166 and Tyr-170. AB - The widely accepted model for toxicity mechanisms of the Bacillus thuringiensis Cry delta-endotoxins suggests that helices alpha4 and alpha5 form a helix-loop helix hairpin structure to initiate membrane insertion and pore formation. In this report, alanine substitutions of two polar amino acids (Asn-166 and Tyr-170) and one charged residue (Glu-171) within the alpha4-alpha5 loop of the 130-kDa Cry4B mosquito-larvicidal protein were initially made via polymerase chain reaction-based directed mutagenesis. As with the wild-type toxin, all of the mutant proteins were highly expressed in Escherichia coli as inclusion bodies upon isopropyl-beta-Dthiogalactopyranoside induction. When E. coli cells expressing each mutant toxin were assayed against Aedes aegypti mosquito larvae, the activity was almost completely abolished for N166A and Y170A mutations, whereas E171A showed only a small reduction in toxicity. Further analysis of these two critical residues by induction of specific mutations revealed that polarity at position 166 and highly conserved aromaticity at position 170 within the alpha4-alpha5 loop play a crucial role in the larvicidal activity of the Cry4B toxin. PMID- 12721493 TI - Cloning, co-expression with an amidating enzyme, and activity of the scorpion toxin BmK ITa1 cDNA in insect cells. AB - BmK ITa1 is an insect-specific neurotoxin from the Chinese scorpion Buthus martensi Karsch (Bmk). We succeeded in obtaining biologically active recombinant BmK ITa1 protein by simultaneous expression in insect cells of BmK ITa1 cDNA with an amidating enzyme expressed by the rat peptidylglycine alpha-amidating monooxygenase (PAM) gene. We investigated the insecticidal efficacy of recombinant BmK ITa1/W (without coexpression of PAM), and of BmK ITa1/A (with coexpression of PAM) in 5th instar Bombyx mori, by injecting these recombinant toxins into larvae. The lethal time for 50% of larvae (LT50) was 9 h for BmK ITa1/A and 17 h for BmK ITa1/W. At 19 h after injection all of the larvae exposed to BmK ITa1/A had been killed, whereas only half of the larvae exposed to BmK ITa1/W had been killed. These results show that the simultaneous expression of an amidating enzyme can result in apparently higher insecticidal activity of BmK ITa1. PMID- 12721494 TI - Historical perspective of the renin-angiotensin system. AB - Researchers continue to be fascinated with the renin-angiotensin system (RAS) more than 100 yr after its discovery because of its powerful role in controlling sodium balance, body fluid volumes, and arterial pressure. Development of drugs that block different components of this system has led to powerful treatments for hypertension, heart failure, diabetes, and other diseases. Molecular approaches to studying this system offer new possibilities for better understanding the physiology and pathophysiology of the RAS, and for developing new therapeutic paradigms. Our challenge in the future will be to effectively utilize the technological advances that are taking place in virtually all areas of science, including the RAS, and to translate them into a better understanding of the pathophysiology and treatment of human diseases. PMID- 12721496 TI - Determination of amino acids in foods by reversed-phase HPLC with new precolumn derivatives, butylthiocarbamyl, and benzylthiocarbamyl derivatives compared to the phenylthiocarbamyl derivative and ion exchange chromatography. AB - New precolumn derivatizing reagents for analysis of amino acids by HPLC butylisothiocyanate (BITC) and benzylisothiocyanate (BZITC)-reacted quantitatively with 22 standard amino acids and the amino acids in the acid hydrolysate of food and protein standard, bovine serum albumin (BSA), at 40 degrees C for 30 min to yield butylthiocarbamyl (BTC) amino acids and at 50 degrees C for 30 min to yield benzylthiocarbammyl (BZTC) amino acids. BTC and BZTC amino acids were successfully separated in 35 min on the reversed-phase Nova Pak C18 column (30 cm x 3.9 mm, 4 microm). The optimum wavelengths for determination of BTC and BZTC derivatives were 240 nm and 246 nm, respectively. Analysis of the results obtained with BSA and food samples as BTC and BZTC derivatives showed good agreement with those determined as ionexchange chromatography and data presented in the literature. The advantage of BITC reagent over the phenylisothiocyanate (PITC) and BZITC was that it had high volatility, so the excess reagent and by-products were easily removed in about 10 min, compared to about 1 h in the PITC and BZITC reagents. In the BTC and BZTC derivatives, cystine and cysteine were determined separately, but in the PTC amino acids derivatized with PITC reagent they were resolved into single peak. PMID- 12721495 TI - The use of capillary electrophoresis for DNA polymorphism analysis. AB - Capillary electrophoresis has advanced enormously over the last 10 yr as a tool for DNA sequencing, driven by the human and other major genome projects and by the need for rapid electrophoresis-based DNA diagnostic tests. The common need of these analyses is a platform providing very high throughput, high-quality data, and low process costs. These demands have led to capillary electrophoresis machines with multiple capillaries providing highly parallel analyses, to new electrophoresis matrices, to highly sensitive spectrofluorometers, and to brighter, spectrally distinct fluorescent dyes with which to label DNA. Capillary devices have also been engineered onto microchip formats, on which both the amount of sample required for analysis and the speed of analysis are increased by an order of magnitude. This review examines the advances made in capillary and chip-based microdevices and in the different DNA-based assays developed for mutation detection and genotype analysis using capillary electrophoresis. The automation of attendant processes such as for DNA sample preparation, PCR, and analyte purification are also reviewed. Together, these technological developments provide the throughput demanded by the large genome-sequencing projects. PMID- 12721497 TI - Assay interference leading to misdiagnosis of central precocious puberty. AB - Immunoassays are widely used to determine hormone levels. Antibodies directed against components of the immunoassay system can interfere with analyte concentration estimates. When unrecognized by clinicians, inappropriate clinical intervention may follow. The case of a young child with premature thelarche and elevated basal and stimulated luteinizing hormone (LH) levels is presented, in whom it is hypothesized that heterophile antibodies (HAs) caused interference in the LH immunoassay. LH concentrations were measured in two different assays: LH microparticle enzyme immunoassay (MEIA) and LH-immunochemiluminometric assay (ICMA). To detect HA interference, LH level was remeasured after both preincubation with mouse serum to neutralize human anti-mouse antibodies, and treatment with a heterophile-blocking tube. The mean basal LH concentration by LH MEIA was 7.4 mIU/mL and for LH-ICMA was 0.08 mIU/mL (normal range for age: 0.02 0.3 mIU/mL). LH concentration by MEIA was 0.08 mIU/mL after preincubation with mouse serum and 2.7 mIU/mL after preincubation with a heterophile blocking tube. In conclusion, HAs were identified in the serum of a child with premature thelarche. The presence of HAs led to spuriously elevated basal and gonadotropin releasing hormone-stimulated LH concentrations, resulting in a diagnosis of central precocious puberty and unnecessary therapy. To avoid similar cases in the future, clinicians should consider the possibility of assay interference when the clinical picture is incongruent with the laboratory data. PMID- 12721499 TI - Effect of metformin and sulfonylurea on C-reactive protein level in well controlled type 2 diabetics with metabolic syndrome. AB - The objective of this study was to examine the effect of the antihyperglycemic agents metformin (insulin sensitizer) and glibenclamide (insulin secretory agent) on the serum level of C-reactive protein (CRP) in well-controlled type 2 diabetics with metabolic syndrome. The participants were diabetic patients being followed in the medical outpatient clinic of King Abdulaziz University Hospital. The inclusion criteria were type 2 diabetics with the metabolic syndrome, well controlled blood glucose on metformin alone or glibenclamide alone, and exclusion of major medical illness. Patients were divided into two groups according to the antihyperglycemic agent used. CRP level was measured 4-wk apart and the mean was calculated. The following data were collected from the study groups: age, sex, body mass index (BMI), duration of diabetes, smoking history, presence of hypertension, hyperlipidemia, and mean CRP level. A total of 110 patients were studied, 65 using metformin and 45 using glibenclamide. CRP level was significantly lower in patients using metformin for blood glucose control compared with those using glibenclamide, 5.56 and 8.3 mg/L, respectively (p = 0.01). A significantly higher level was observed in hypertensive and hyperlipidemic patients compared with normotensive and normolipidemic, 5.3 vs 3.2 mg/L and 7.1 vs 4.3 mg/L, respectively (p = 0.02, 0.01). There was a statistically significant correlation between CRP and BMI (r = 0.37) and age (r = 0.36) (all p = 0.01). The data showed that metformin decreases the level of circulating CRP, a marker of inflammation, more than glibenclamide. PMID- 12721500 TI - Expression of membrane prostaglandin E synthase in human placenta and fetal membranes and effect of labor. AB - Initiation and maintenance of labor in humans is associated with an increase in prostaglandin synthesis by intrauterine tissues. The objective of the present study was to characterize the distribution of membrane-bound PGES (mPGES) protein and mPGES mRNA in human placenta, fetal membranes, and decidua at term and to determine whether any changes occurred with labor. Immunoreactive mPGES was found to be highly concentrated in amnion epithelial cells and the chorion laeve trophoblasts, with lower levels in the mesenchymal layers. The enzyme was at very low levels or undetectable in the decidual tissue. Much lower levels of mPGES protein and mRNA were found in placenta than in fetal membranes. mPGES was associated with the syncytiotrophoblast and in cells surrounding blood vessels. The expression of mPGES mRNA did not change with labor in full membranes or placenta, but Western analysis showed an increase in mPGES protein in chorion laeve and a decrease in mPGES protein in placenta during labor, with no change in the amnion. The differences in expression found among placenta, chorion, and amnion before and after labor would indicate that this enzyme is differentially regulated in these tissues at this time. PMID- 12721498 TI - Neuropeptides of the pituitary adenylate cyclase-activating polypeptide/vasoactive intestinal polypeptide/growth hormone-releasing hormone/secretin family in testis. AB - Mammalian testicular development and the maintenance of spermatogenesis are hormone-dependent processes that are controlled by the pituitary gonadotropins and testosterone. Recent studies have demonstrated the presence of many neuropeptides and their receptors in the testis, suggesting that these peptides operate as local regulators of testicular germ cell development and function. Among these testicular neuropeptides, the peptides that belong to the pituitary adenylate cyclase-activating polypeptide (PACAP) family, particularly growth hormone-releasing hormone and secretin, appear to show some unique common features in terms of intratesticular localization and the time of expression during the spermatogenic cycle. However, their precise physiologic roles and mechanisms of action remain unknown. This review analyzes the available information on the functional interactions among the testicular cells that appear to be mediated by locally produced neuropeptides, with a special emphasis on the peptides of the PACAP family. PMID- 12721501 TI - Effect of 17beta-estradiol on mRNA expression of large- conductance, voltage dependent, and calcium-activated potassium channel alpha and beta subunits in guinea pig. AB - Large-conductance, voltage- and calcium-activated potassium (MaxiK) channels play a key role in cell excitability. MaxiK channels are composed of a pore-forming alpha-subunit and a regulatory beta-subunit, of which four (beta1-4) genes have been identified. Previous findings suggested that MaxiK channel activity is regulated by estradiol. However, the underlying mechanisms have remained incompletely documented. Therefore, we used reverse transcriptase polymerase chain reaction to clone four cDNA fragments that were specific to the guinea pig alpha, beta1, beta2, and beta4 genes. Using a sensitive ribonuclease protection assay, we found that the alpha and beta4 mRNAs were the most abundant mRNAs in the brain and pituitary, whereas in the aorta, the alpha-subunit was coexpressed with the beta1-subunit. Moreover, there was a significant upregulation of the alpha- but not the beta1-subunit mRNA and the alpha-subunit protein in the aorta of the estrogenvs oil-treated ovariectomized animals. In specific brain areas including preoptic area, ventral hypothalamus, hippocampus, and amygdala, and in the pituitary, neither the alpha- nor beta4-subunit mRNAs were affected by estrogen. These findings suggest that estrogen may not affect the mRNA expression of MaxiK channels in the brain and pituitary. However, estrogen causes increased expression of MaxiK alpha in the aorta, which may explain some of the cardioprotective effects of estrogen in women. PMID- 12721502 TI - Impact of estradiol on parametrial adipose tissue function: evidence for establishment of a new set point of leptin sensitivity in control of energy metabolism in female rat. AB - Estradiol has been implicated in the regulation of food intake; however, its effect seems to be exerted in a bimodal fashion. We examined whether a single im injection of estradiol valerate (E2V), lastingly effective, could induce changes in parametrial fat function that further induce a new set point of leptin sensitivity in the female rat. E2V induced severe anorexia and loss of body weight between d 4 and 12 posttreatment. E2V rats recovered normal food intake and departing body weights on wk 2 and 3 posttreatment, respectively; however, they did not reach body weights of control rats. On d 61 posttreatment, we found that unfasting E2V, vs control, rats displayed increased E2 and leptin circulating levels; reduced plasma tumor necrosis factor-alpha(TNF-alpha) concentrations; similar circulating levels of glucose, insulin, and triglyceride; and lower parametrial fat mass containing a higher number of adipocytes that, although normal in size, in vitro released more leptin. Metabolic responses to fasting indicated that unlike control animals, E2V rats did not decrease triglyceride circulating levels, and that both groups decreased plasma glucose, leptin, and insulin, but not TNF-alpha, levels. High glucose load experiments indicated that E2V animals displayed a better insulin sensitivity than control rats; did not significantly increase circulating leptin concentrations as control rats did; and, unlike control, significantly decreased plasma triglyceride levels. Our data strongly support a potent acute anorectic effect of E2 and that, after several weeks, E2 modified parametrial fat function and insulin sensitivity, protecting the organism against future unfavorable metabolic conditions. PMID- 12721503 TI - Exploring levels of hexosamine biosynthesis pathway intermediates and protein kinase C isoforms in muscle and fat tissue of Zucker Diabetic Fatty rats. AB - Many studies suggest that insulin resistance develops and/or is maintained by an increased flux of glucose through the hexosamine biosynthesis pathway. This pathway may attenuate insulin-stimulated glucose uptake by activating protein kinase C (PKC). Therefore, we investigated whether the concentrations of the major hexosamine metabolites, uridine diphosphate- N-acetyl-glucosamine (UDP GlcNAc) and uridine diphosphate- N-acetyl-galactosamine (UDP-GalNAc), and the expression levels of PKC isoforms were affected in Zucker Diabetic Fatty (ZDF) rats, an animal model widely used to study type 2 diabetes mellitus. At the age of 6 wk, control and ZDF rats were normoglycemic. Whereas control rats remained normoglycemic, the ZDF rats became hyperglycemic. The amount of UDP-GlcNAc and UDP-GalNAc in muscle tissue of ZDF rats was similar at 6, 12, 18, and 24 wk of age. Moreover, the concentration of both hexosamines did not differ among ZDF, phlorizin-treated ZDF, and control rats. Western blot analysis revealed that PKCalpha, delta, epsilon, andzeta, but not PKCbeta and gamma, were expressed in muscle and fat tissues from 6- and 24-wk-old control and ZDF rats. In addition, we did not observe changes in the expression levels of the PKC isoforms following prolonged hyperglycemia. Taken together, these findings indicate that the amounts of several metabolites from the hexosamine biosynthesis pathway and PKC isoforms, both hypothesized to be important in the development and/or maintenance of the insulin-resistant state of muscle and fat tissue, are not different in ZDF compared with nondiabetic rats. PMID- 12721506 TI - Somatostatin receptors in pituitary and development of somatostatin receptor subtype-selective analogs. AB - Somatostatin receptor (SSTR) subtypes 1, 2, and 5 are expressed in the normal human pituitary. SSTR2 and SSTR5 are expressed in almost all growth hormone (GH) cell adenomas, and prolactin (PRL)-secreting tumors express SSTR5 more than SSTR2. SSTR4 is not detected in all pituitary adenoma subtypes, and SSTR1 and SSTR3 are expressed in about 50% of tumors. Human GH is regulated through ligand binding to both SSTR2 and SSTR5, but octreotide and lanreotide, the two clinically available somatostatin analogs, bind to human SSTR2 much better than to SSTR5. Novel SSTR2- and SSTR5- selective analogs with improved binding affinity for these receptor subtypes are highly potent in suppressing GH release from cultures of human fetal pituitaries or GH-cell adenomas. Only SSTR5 selective analogs suppress in vitro PRL secretion from cultured prolactinomas. A new SSTR2+5 bispecific analog with high affinity and selectivity for both SSTR2 and SSTR5, and a somatostatin analog with a unique broad receptor (SSTR1, 2, 3, and 5) binding profile, are both able to inhibit in vitro GH release in GH cell adenomas partially sensitive to octreotide. Recently, a somatostatindopamine hybrid molecule was introduced with potentially functional synergy on GH and PRL release. Using the expanding knowledge on SSTRs and their ligand activation, the development of novel pharmacologic concepts may open new opportunities for effective manipulation of this complex intracellular signaling system. These concepts may achieve better control of pituitary hormone hypersecretion, pituitary size, as well as antitumor effects in patients with SSTR-expressing tumors. PMID- 12721505 TI - Somatostatin and somatostatin receptor physiology. AB - Since the discovery of somatostatin (SST) over three decades ago, its ubiquitous distribution and manifold functions are still being documented. SST is synthesized in the hypothalamus and transported to the anterior pituitary gland where it tonicaly inhibits GH and TSH secretion as well as being responsible for GH pulsatile release. Several internal feedback loops, sleep, exercise, and chemical agents control and influence SST release. SST also impacts the function of a wide variety of cells and organ systems throughout the body. Knowledge of the structures of the SSTs has resulted in recognition of the essential four core conserved residues responsible for their actions. The SSTs act through six separate SST cell surface receptors (SSTRs), members of the family of G protein coupled receptors. Receptor ligand binding (SST/SSTR) results in cellular activities specific for each receptor, or receptor combinations, and their tissue/cell localization. Understanding the structure/function relationship of the SSTs and their receptors, including the internalization of SST/SSTR complexes, has facilitated the development of a variety of novel pharmacologic agents for the diagnosis and treatment of neuroendocrine tumors and unfolding new applications. PMID- 12721508 TI - Somatostatin analogs in treatment of non-growth hormone-secreting pituitary adenomas. AB - Besides well-known effects in GH-secreting adenomas, somatostatin analogs such as octreotide and lanreotide have been used in TSH-secreting adenomas and in the so called clinically nonfunctioning adenomas. The rationale for their use is based on the evidence that both these tumor types express large amounts of somatostatin receptor subtypes 2 and 5, which are preferentially bound by octreotide and lanreotide. However, whether in TSH-secreting adenomas the results are excellent in the nonfunctioning type, the results are controversial. Some preliminary results showing a very rapid recovery of the visual field have not been confirmed subsequently. No evident effect of tumor shrinkage has been reported. At present, the use of somatostatin analogs in clinically nonfunctioning adenomas is questioned. PMID- 12721507 TI - Somatostatin analogs in medical treatment of acromegaly. AB - Although acromegaly remains a disease primarily addressed by pituitary microsurgery, most patients require secondary treatment for persistent growth hormone (GH) hypersecretion and elevated serum insulin-like growth factor-1 (IGF 1) concentrations following adenomectomy. Persistently abnormal serum GH and IGF 1 can be reduced to normal concentrations in better than half of post-surgery acromegalics using the pharmacologic treatments available at present, the dopamine agonists (DA) and somatostatin (SST) analogs. The long-acting SST analogs octreotide LAR and lanreotide SR have become the mainstay of medical treatment for acromegaly, having largely supplanted DA agents since the introduction of bromocriptine for the suppression of GH secretion in the 1970s. The DA cabergoline may be effective in up to half of patients, however, in particular those patients whose tumors cosecrete prolactin. On the horizon is the GH-receptor antagonist pegvisomant, which is expected to enable the reduction of serum IGF-1 to the normal range in the vast majority of postoperative acromegaly patients, representing a revolutionary development in the medical treatment of this disease. We here review the choices available to the endocrinologist in the pharmacologic treatment of acromegaly, focusing upon the SST analogs. PMID- 12721510 TI - Somatostatin analogs as primary medical therapy for acromegaly. AB - Acromegaly is a debilitating disease usually caused by a growth-hormone secreting pituitary adenoma. Therapeutic goals include improvement of symptoms, reduction in tumor mass, biochemical normalization, and preservation of pituitary function. Treatment options include transsphenoidal surgery, radiation, and pharmacotherapy. In view of the good cure rate, surgery remains the therapeutic modality of choice for most patients with microadenomas or well-circumscribed macroadenomas. In contrast, >40% of patients with invasive macroadenomas (who make up the majority of patients with acromegaly) will have residual disease following surgery, and require additional therapeutic intervention. Somatostatin analogs result in biochemical normalization in >60% of non-operated patients, and are well tolerated. Therefore, somatostatin analogs have emerged as a rational first-line treatment for the appropriately selected patient with acromegaly. PMID- 12721509 TI - Somatostatin analog therapy in treatment of gastrointestinal disorders and tumors. AB - Long-acting octapeptide somatostatin analogs can effectively control symptoms resulting from excessive hormone release in patients with endocrine tumors of the gastrointestinal tract, provided that these tumors and metastases show a high expression of the somatostatin receptor subtype 2. The presence of this receptor subtype on these tumors can be demonstrated by in vitro studies, but also in vivo using 111In-pentetreotide scintigraphy. In a few studies, significant antiproliferative effects of these drugs on these tumors have also been demonstrated. The effectiveness of octapeptide somatostatin analogs in the management of chemotherapy- related and AIDS-related diarrhea and in reducing postoperative complications of pancreatic surgery have also been demonstrated. These drugs have been used to decrease the output of enterocutaneous pancreatic fistulas and are prophylactically used to prevent the development of these fistulas. Octapeptide somatostatin analog therapy is widely accepted for the initial management of acute variceal bleeding in cirrhotic patients. These drugs are currently also being evaluated for the treatment of advanced hepatocellular carcinoma and malignant intestinal obstruction. Radiotherapy with octapeptide somatostatin analogs coupled to radionuclides such as indium-111, yttrium-90, and lutetium- 177 is currently being studied in phase I-III trials. PMID- 12721511 TI - Comparison of efficacy and tolerability of somatostatin analogs and other therapies for acromegaly. AB - Medical therapy plays an important role in the management of acromegaly. Dopamine agonists and somatostatin analogs are two classes of drugs approved for this purpose worldwide. Pegvisomant, a growth hormone receptor antagonist, has recently been evaluated in clinical trials. Somatostatin analogs have been the mainstay of medical treatment during the last 10 yr with their acceptability enhanced by the development of depot preparations. Somatostatin analogs improve symptoms and signs of acromegaly in the majority, normalize IGF- 1 in up to 60%, and result in tumor shrinkage in up to half of patients. Dopamine agonists have modest efficacy and limited tolerability. They are more effective in mixed GH/prolactin-secreting tumors. Newer agonists with D2 receptor specificity have fewer side effects but are less efficacious than somatostatin analogs. The addition of a dopamine agonist to somatostatin analog therapy can result in greater biochemical control than with individual agents. Pegvisomant is the most effective drug treatment for acromegaly, but it is likely to have a major adjuvant role as its mechanism of action is not directed at the tumor. The availability of more effective and better tolerated drug therapies offers greater flexibility and individualization of therapy that will lead to improved patient care and disease control. PMID- 12721512 TI - Octreoscan radioreceptor imaging. AB - With the in vivo demonstration of somatostatin-receptor- positive tumors in patients using a radiolabeled somatostatin analog, peptide receptor scintigraphy became available some 15 yr ago. Octreoscan is a radiopharmaceutical with proven clinical importance in the visualization of somatostatin-receptor-positive tumors, and the overall sensitivity of somatostatin receptor imaging is high. In a number of neuroendocrine tumor types, as well as in Hodgkin's disease, inclusion of somatostatin receptor imaging in the localization or staging procedure may be very rewarding. The value of somatostatin receptor imaging in patients with other tumors, like breast cancer, or in patients with granulomatous diseases, has to be established. PMID- 12721513 TI - Baculoviruses as Vectors for Gene Therapy against Human Prostate Cancer. AB - Current curative strategies for prostate cancer are restricted to the primary tumour, and the effect of treatments to control metastatic disease is not sustained. Therefore, the application of gene therapy to prostate cancer is an attractive alternative. Baculoviruses are highly restricted insect viruses, which can enter, but not replicate in mammalian cells. Baculoviruses can incorporate large amounts of extra genetic material, and will express transgenes in mammalian cells when under the control of a mammalian or strong viral promoter. Successful gene delivery has been achieved both in vitro and in vivo and into both dividing and nondividing cells, which is important since prostate cancers divide relatively slowly. In addition, the envelope protein gp64 is sufficiently mutable to allow targeted transduction of particular cell types. In this review, the advantages of using baculoviruses for prostate cancer gene therapy are explored, and the mechanisms of viral entry and transgene expression are described. PMID- 12721514 TI - In Vivo Noninvasive Imaging for Gene Therapy. AB - Gene therapy is reaching a stage where some clinical benefits have been demonstrated on patients involved in phase I/II clinical trials. However, in many cases, the clinical benefit is hardly measurable and progress in the improvement of gene therapy formulations is hampered by the lack of objective clinical endpoints to measure transgene delivery and to quantitate transgene expression. However, these endpoints rely almost exclusively on the analysis of biopsies by molecular and histopathological methods. These methods provide only a limited picture of the situation. Therefore, there is a need for a technology that would allow precise, spacio-temporal measurement of gene expression on a whole body scale upon administration of the gene delivery vector. In the field of gene therapy, a considerable effort is being invested in the development of noninvasive imaging of gene expression and this review presents the various strategies currently being developed. PMID- 12721515 TI - Targeting Radiotherapy to Cancer by Gene Transfer. AB - Targeted radionuclide therapy is an alternative method of radiation treatment which uses a tumor-seeking agent carrying a radioactive atom to deposits of tumor, wherever in the body they may be located. Recent experimental data signifies promise for the amalgamation of gene transfer with radionuclide targeting. This review encompasses aspects of the integration of gene manipulation and targeted radiotherapy, highlighting the possibilities of gene transfer to assist the targeting of cancer with low molecular weight radiopharmaceuticals. PMID- 12721517 TI - Gene Therapy for Cardiovascular Disease. AB - The last decade has seen substantial advances in the development of gene therapy strategies and vector technology for the treatment of a diverse number of diseases, with a view to translating the successes observed in animal models into the clinic. Perhaps the overwhelming drive for the increase in vascular gene transfer studies is the current lack of successful long-term pharmacological treatments for complex cardiovascular diseases. The increase in cardiovascular disease to epidemic proportions has also led many to conclude that drug therapy may have reached a plateau in its efficacy and that gene therapy may represent a realistic solution to a long-term problem. Here, we discuss gene delivery approaches and target diseases. PMID- 12721520 TI - Charles Dickens and the movement for sanitary reform. AB - Charles Dickens's adult life parallels the period when the movement for sanitary reform took root in England. Although he was not one of its leaders, he became in time one of its most outspoken advocates. This essay describes Dickens's growing involvement in the sanitary movement and looks at one of the most important ways he supported it--articles published in his weekly journal Household Words PMID- 12721519 TI - Evolution by phenotype: a biomedical perspective. AB - Genes are widely assumed to play a major role in the epidemiology of complex chronic diseases, yet attempts to characterize the genetic architecture of such traits have been frustrating. Understanding that evolution works by screening phenotypes rather than genotypes can help explain the source of this frustration. Complex traits are usually the result of long-term, often subtle, gene environment interactions, such that individual life histories may be as important as population histories in predicting and explaining these traits. Recognizing that the problem is not due to technological limitations can help temper expectations and guide the design of future work in biomedical genetics, by allowing us to focus on better approaches where they exist and on those problems most likely to yield a genetic solution. We may even be forced to re-conceive complex biological causation. PMID- 12721521 TI - Animals, disease, and man: making connections. AB - The intricate causal relationships between disease in man and disease in animals first began to be elucidated in the mid-19th century. Although the connections between animal and human disease are now generally understood, individuals as well as societies remain slow to act on this knowledge. This paper examines the gradual recognition of these disease connections and explores the parallel theme of man's reluctance to appreciate the implications of these connections. It identifies factors that have inhibited the realization of the links between disease in man and animals, and discusses several milestones in the scientific elucidation of these links. Beginning with emerging concerns over the relationship between bovine and human tuberculosis in the 1860s, it follows the discovery of insect vectors, animal reservoirs, and the links between animals, influenza, and man. Despite warnings of the potential significance for human disease of patterns of changes in the relationship with animals and the natural world, scientists have continued to treat human and animal health as largely independent disciplines, while historians too have neglected this important aspect of human disease. PMID- 12721518 TI - Targeting of Synthetic Gene Delivery Systems. AB - Safe, efficient, and specific delivery of therapeutic genes remains an important bottleneck for the development of gene therapy. Synthetic, nonviral systems have a unique pharmaceutical profile with potential advantages for certain applications. Targeting of the synthetic vector improves the specificity of gene medicines through a modulation of the carriers' biodistribution, thus creating a dose differential between healthy tissue and the target site. The biodistribution of current carrier systems is being influenced to a large extent by intrinsic physicochemical characteristics, such as charge and size. Consequently, such nonspecific interactions can interfere with specific targeting, for example, by ligands. Therefore, a carrier complex should ideally be inert, that is, free from intrinsic properties that would bias its distribution away from the target site. Strategies such as coating of DNA carrier complexes with hydrophilic polymers have been used to mask some of these intrinsic targeting effects and avoid nonspecific interactions. Preexisting endogenous ligand-receptor interactions have frequently been used for targeting to certain cell types or tumours. Recently exogenous ligands have been derived from microorganisms or, like antibodies or phage-derived peptides, developed de novo. In animal models, such synthetic vectors have targeted remote sites such as a tumour. Furthermore, the therapeutic proof of the concept has been demonstrated for fitting combinations of synthetic vectors and therapeutic gene. PMID- 12721522 TI - Rethinking the boundaries of Kawasaki disease: toward a revised case definition. AB - This paper describes the historical evolution of the Kawasaki disease (KD) case definition and its limitations for identification and treatment of children at risk for coronary artery aneurysms (CAA). The dominant view of pathogenesis is that an unknown agent infects infants and children, who then develop the signs of KD. Some of the infected infants and children then develop CAA, and a few die from myocardial infarction. Because the etiologic agent remains unknown, diagnosis of KD relies on observation and recognition of the clinical signs that comprise the KD case definition criteria. This approach has been successful in identifying and treating many children at risk for CAA. Unfortunately, however, it has delayed the effective treatment of children who fail to meet the KD case definition criteria but who, nevertheless, develop CAA. The original case definition was developed before the general acceptance of CAA as sequelae of KD, the availability of the echocardiogram, and effective treatment with intravenous immunoglobulin. Despite an evolution in awareness, detection, and treatment of possible CAA sequela, the case definition has not been altered so as to incorporate this knowledge. Our investigation explores the transformation of the case definition from an epidemiological instrument to a diagnostic tool. We urge the construction of a more sensitive KD case definition that includes signs and laboratory findings associated with CAA. PMID- 12721523 TI - Scholars, investigators, and entrepreneurs: the metamorphosis of American medicine. PMID- 12721516 TI - Transcriptional Targeting in Cancer Gene Therapy. AB - Cancer gene therapy has been one of the most exciting areas of therapeutic research in the past decade. In this review, we discuss strategies to restrict transcription of transgenes to tumour cells. A range of promoters which are tissue-specific, tumour-specific, or inducible by exogenous agents are presented. Transcriptional targeting should prevent normal tissue toxicities associated with other cancer treatments, such as radiation and chemotherapy. In addition, the specificity of these strategies should provide improved targeting of metastatic tumours following systemic gene delivery. Rapid progress in the ability to specifically control transgenes will allow systemic gene delivery for cancer therapy to become a real possibility in the near future. PMID- 12721524 TI - Physicians and the pharmaceutical industry: a dysfunctional relationship. AB - The pharmaceutical industry is one of the largest and most profitable industries in the world, and in the United States, the industry has a particularly privileged economic position. Yet the cost of drugs in the United States is higher than anywhere else, due largely to the fact that the industry is focusing increasingly on marketing rather than on the development of meaningful new medications: available evidence does not support claims of great expense for the development of new drugs. Because of its vast resources, the pharmaceutical industry has assumed an increasing influence in medicine, which, given the differences in values and priorities between medicine and the drug companies, is a cause for concern. The pharmaceutical industry has acted to maximize its profits in ways that frequently conflict with medicine's need for truth and full disclosure. Indeed, the industry has arguably worked to compromise physicians' judgments, as well as academic standards. As a result, despite government regulation there have been unnecessary adverse effects from drugs. The experience with butorphanol (Stadol) exemplifies problems in the current system and the harm that can result. Changes are suggested to make the pharmaceutical industry more responsive to the needs of patients and physicians. PMID- 12721525 TI - Where have all the giants gone? Reconciling medical education and the traditions of patient care with limitations on resident work hours. AB - The Accreditation Council for Graduate Medical Education recently approved regulations that would prohibit residents from working more than 80 hours per week and more than 24 hours at a stretch. These regulations are scheduled to take effect in all U.S. teaching hospitals on 1 July 2003. Those who approve of the proposed regulations argue that house staff fatigue is responsible for physician error, depression, anger, and a lack of compassion for patients. But critics point to the adverse effects on key goals of house staff training--the development of accountability and responsibility. Can the rigorous discipline of medical education and the long tradition of medicine as a profession be reconciled with the current calls for limiting resident duty hours and on-call schedules? The intensity of patient care in teaching hospitals today is far greater than it was in the past. These changes in medical care make it critical to develop new programs that will reconcile rigorous, scientifically based humanistic medicine with the needs of patients and physicians. This will require imaginative and creative solutions that take a larger view of medical education and medical care than mere manpower calculations and numerical solutions focused simply on compliance with an 80-hour work week. PMID- 12721526 TI - We-intentionality: an essential element in understanding human culture? AB - Culture is a powerful cause of human behavior, yet there have been few attempts to synthesize the biological and social sciences in order to provide an explanation of culture and hence to lay the foundations for a comprehensive human science. This essay explores one possible way of doing this by outlining the likely psychological mechanisms that give rise to the unique human capacity for creating social reality. The core assumption is that "we-intentionality," which is key to understanding social constructions, can and must be linked to specific evolved psychological traits, and hence placed within a neuroscience perspective. PMID- 12721527 TI - Growth and photosynthesis of Japanese flowering cherry under simulated microgravity conditions. AB - The photosynthetic rate, the leaf characteristics related to photosynthesis, such as the chlorophyll content, chlorophyll a/b ratio and density of the stomata, the leaf area and the dry weight in seedlings of Japanese flowering cherry grown under normal gravity and simulated microgravity conditions were examined. No significant differences were found in the photosynthetic rates between the two conditions. Moreover, leaf characteristics such as the chlorophyll content, chlorophyll a/b ratio and density of the stomata in the seedlings grown under the simulated microgravity condition were not affected. However, the photosynthetic product of the whole seedling under the simulated microgravity condition increased compared with the control due to its leaf area increase. The results suggest that dynamic gravitational stimulus controls the partitioning of the products of photosynthesis. PMID- 12721528 TI - [Perspective on gravitational biology of amphibians]. AB - We review here the scientific significance of the use of amphibians for research in gravitational biology. Since amphibian eggs are quite large, yet develop rapidly and externally, it is easy to observe their development. Consequently amphibians were the first vertebrates to have their early developmental processes investigated in space. Though several deviations from normal embryonic development occur when amphibians are raised in microgravity, their developmental program is robust enough to return the organisms to an ostensibly normal morphology by the time they hatch. Evolutionally, amphibians were the first vertebrate animal to come out of the water and onto land. Subsequently they diversified and have adaptively radiated to various habitats. They now inhabit aquatic, terrestrial, arboreal and fossorial niches. This diversity can be used to help study the biological effects of gravity at the organismal level, where macroscopic phenomena are associated with gravitational loading. By choosing different amphibian models and using a comparative approach one can effectively identify the action of gravity on biological systems, and the adaptation that vertebrates have made to this loading. Advances in cellular and molecular biology provide powerful tools for the study in many fields, including gravitational biology, and amphibians have proven to be good models for studies at those levels as well. The low metabolic rates of amphibians make them convenient organisms to work with (compared to birds and mammals) in the difficult and confined spaces on orbiting research platforms. We include here a review of what is known about and the potential for further behavioral and physiological researches in space using amphibians. PMID- 12721540 TI - From the ground up. PMID- 12721541 TI - Conjugal rites of mycobacteria. PMID- 12721543 TI - Centrosomes as DNA damage regulators. PMID- 12721542 TI - A nuclear address with influence. PMID- 12721544 TI - ORFeomics: correcting the wiggle in worm genes. PMID- 12721547 TI - The human phenome project. PMID- 12721548 TI - Genomic analysis of primary tumors does not address the prevalence of metastatic cells in the population. PMID- 12721549 TI - Genetic background is an important determinant of metastatic potential. PMID- 12721554 TI - Recipe for disaster. PMID- 12721553 TI - Assessing the impact of biobanks. PMID- 12721555 TI - Concerns over refuge size for US EPA-approved Bt corn. PMID- 12721556 TI - Europe sees sharp decline in GMO research. PMID- 12721557 TI - US bioterrorism countermeasures research builds across a broad front. PMID- 12721558 TI - US FDA approves new class of HIV therapeutics. PMID- 12721560 TI - Thermodynamics--old laws in medicine and complex disease. PMID- 12721561 TI - Will we reap what biopharming sows? PMID- 12721562 TI - The set of embryo subjects. PMID- 12721563 TI - Splurge or merge? PMID- 12721565 TI - Organelle evolution meets biotechnology. PMID- 12721566 TI - Global architecture of genetic interactions on the protein network. PMID- 12721567 TI - Developing countries and systems biology. PMID- 12721568 TI - A genome-wide view of antisense. PMID- 12721569 TI - Delivering zinc fingers. PMID- 12721570 TI - GMO opposition not based on a mistake. PMID- 12721571 TI - Walking the drug regulatory tightrope. PMID- 12721572 TI - Biobusiness on the Web. PMID- 12721573 TI - The Streptomyces genome--be prepared! PMID- 12721574 TI - Engineering what comes naturally. PMID- 12721575 TI - Membrane proteins ride shotgun. PMID- 12721576 TI - The rollercoaster ride to anti-cancer antibodies. PMID- 12721578 TI - Integrating intellectual property within the organizational social structure. PMID- 12721583 TI - A chance for change in France. PMID- 12721582 TI - The past, present, and future of biotechnology in Mexico. PMID- 12721584 TI - Rice institute needs strong support. PMID- 12721586 TI - Biologists seek to head off future sources of infection. PMID- 12721587 TI - Critics slam treatment for SARS as ineffective and perhaps dangerous. PMID- 12721588 TI - Neutron source powers ahead with weapons-grade uranium. PMID- 12721585 TI - Apartment complex holds clues to pandemic potential of SARS. PMID- 12721590 TI - Commission lays foundations for rise in research spending. PMID- 12721589 TI - Human fatality adds fresh impetus to fight against bird flu. PMID- 12721591 TI - French researchers demand radical overhaul of funding. PMID- 12721592 TI - Arizona institute names leader. PMID- 12721593 TI - Columbia inquiry homes in on faulty foam. PMID- 12721594 TI - Stealth ship sets sail for a quiet life fishing for data. PMID- 12721596 TI - Cell nanotechnology: The tiny toolkit. PMID- 12721597 TI - Brazilian forensic medicine: Back from the dead. PMID- 12721599 TI - Oil and war: we had the warning 30 years ago. PMID- 12721598 TI - Improving science through online commentary. PMID- 12721604 TI - Innate immunity: The unsung heroes. PMID- 12721605 TI - Structural biology: Life's transistors. PMID- 12721606 TI - Optics: Positively negative. PMID- 12721608 TI - Genomics: Relative pathogenic values. PMID- 12721609 TI - Earth science: Subduction the hard way. PMID- 12721610 TI - Chemical physics: How to keep dry in water. PMID- 12721611 TI - Molecular biology: Complicity of gene and pseudogene. PMID- 12721612 TI - Genetics: Suicidal mushroom cells. PMID- 12721613 TI - Atmospheric chemistry: Burning domestic issues. PMID- 12721614 TI - Astronomy: Elements of surprise. PMID- 12721616 TI - Insect communication: Polarized light as a butterfly mating signal. PMID- 12721617 TI - Social insects: Cuticular hydrocarbons inform task decisions. PMID- 12721618 TI - X-ray structure of a voltage-dependent K+ channel. AB - Voltage-dependent K+ channels are members of the family of voltage-dependent cation (K+, Na+ and Ca2+) channels that open and allow ion conduction in response to changes in cell membrane voltage. This form of gating underlies the generation of nerve and muscle action potentials, among other processes. Here we present the structure of KvAP, a voltage-dependent K+ channel from Aeropyrum pernix. We have determined a crystal structure of the full-length channel at a resolution of 3.2 A, and of the isolated voltage-sensor domain at 1.9 A, both in complex with monoclonal Fab fragments. The channel contains a central ion-conduction pore surrounded by voltage sensors, which form what we call 'voltage-sensor paddles' hydrophobic, cationic, helix-turn-helix structures on the channel's outer perimeter. Flexible hinges suggest that the voltage-sensor paddles move in response to membrane voltage changes, carrying their positive charge across the membrane. PMID- 12721619 TI - The principle of gating charge movement in a voltage-dependent K+ channel. AB - The steep dependence of channel opening on membrane voltage allows voltage dependent K+ channels to turn on almost like a switch. Opening is driven by the movement of gating charges that originate from arginine residues on helical S4 segments of the protein. Each S4 segment forms half of a 'voltage-sensor paddle' on the channel's outer perimeter. Here we show that the voltage-sensor paddles are positioned inside the membrane, near the intracellular surface, when the channel is closed, and that the paddles move a large distance across the membrane from inside to outside when the channel opens. KvAP channels were reconstituted into planar lipid membranes and studied using monoclonal Fab fragments, a voltage sensor toxin, and avidin binding to tethered biotin. Our findings lead us to conclude that the voltage-sensor paddles operate somewhat like hydrophobic cations attached to levers, enabling the membrane electric field to open and close the pore. PMID- 12721620 TI - Ligand-receptor binding revealed by the TNF family member TALL-1. AB - The tumour necrosis factor (TNF) ligand TALL-1 and its cognate receptors, BCMA, TACI and BAFF-R, were recently identified as members of the TNF superfamily, which are essential factors contributing to B-cell maturation. The functional, soluble fragment of TALL-1 (sTALL-1) forms a virus-like assembly for its proper function. Here we determine the crystal structures of sTALL-1 complexed with the extracellular domains of BCMA and BAFF-R at 2.6 and 2.5 A, respectively. The single cysteine-rich domain of BCMA and BAFF-R both have saddle-like architectures, which sit on the horseback-like surface formed by four coil regions on each individual sTALL-1 monomer. Three novel structural modules, D2, X2 and N, were revealed from the current structures. Sequence alignments, structural modelling and mutagenesis revealed that one disulphide bridge in BAFF R is critical for determining the binding specificity of the extracellular domain eBAFF-R to TALL-1 instead of APRIL, a closely related ligand of TALL-1, which was confirmed by binding experiments in vitro. PMID- 12721621 TI - The elemental abundance pattern in a galaxy at z = 2.626. AB - The discovery of metal-poor stars (where metal is any element more massive than helium) has enabled astronomers to probe the chemical enrichment history of the Milky Way. More recently, element abundances in gas inside high-redshift galaxies has been probed through the absorption lines imprinted on the spectra of background quasars, but these have typically yielded measurements of only a few elements. Furthermore, interpretation of these abundances is complicated by the fact that differential incorporation of metals into dust can produce an abundance pattern similar to that expected from nucleosynthesis by massive stars. Here we report the observation of over 25 elements in a galaxy at redshift z = 2.626. With these data, we can examine nucleosynthetic processes independent of the uncertainty arising from depletion. We find that the galaxy was enriched mainly by massive stars (M > 15 solar masses) and propose that it is the progenitor of a massive elliptical galaxy. The detailed abundance patterns suggest that boron is produced through processes that act independently of metallicity, and may require alternative mechanisms for the nucleosynthesis of germanium. PMID- 12721622 TI - Interplanetary dust from the explosive dispersal of hydrated asteroids by impacts. AB - The Earth accretes about 30,000 tons of dust particles per year, with sizes in the range of 20-400 microm (refs 1, 2). Those particles collected at the Earth's surface--termed micrometeorites--are similar in chemistry and mineralogy to hydrated, porous meteorites, but such meteorites comprise only 2.8% of recovered falls. This large difference in relative abundances has been attributed to 'filtering' by the Earth's atmosphere, that is, the porous meteorites are considered to be so friable that they do not survive the impact with the atmosphere. Here we report shock-recovery experiments on two porous meteorites, one of which is hydrated and the other is anhydrous. The application of shock to the hydrated meteorite reduces it to minute particles and explosive expansion results upon release of the pressure, through a much broader range of pressures than for the anhydrous meteorite. Our results indicate that hydrated asteroids will produce dust particles during collisions at a much higher rate than anhydrous asteroids, which explains the different relative abundances of the hydrated material in micrometeorites and meteorites: the abundances are established before contact with the Earth's atmosphere. PMID- 12721623 TI - Bose-Einstein condensation of the triplet states in the magnetic insulator TlCuCl3. AB - Bose-Einstein condensation denotes the formation of a collective quantum ground state of identical particles with integer spin or intrinsic angular momentum. In magnetic insulators, the magnetic properties are due to the unpaired shell electrons that have half-integer spin. However, in some such compounds (KCuCl3 and TlCuCl3), two Cu2+ ions are antiferromagnetically coupled to form a dimer in a crystalline network: the dimer ground state is a spin singlet (total spin zero), separated by an energy gap from the excited triplet state (total spin one). In these dimer compounds, Bose-Einstein condensation becomes theoretically possible. At a critical external magnetic field, the energy of one of the Zeeman split triplet components (a type of boson) intersects the ground-state singlet, resulting in long-range magnetic order; this transition represents a quantum critical point at which Bose-Einstein condensation occurs. Here we report an experimental investigation of the excitation spectrum in such a field-induced magnetically ordered state, using inelastic neutron scattering measurements of TlCuCl3 single crystals. We verify unambiguously the theoretically predicted gapless Goldstone mode characteristic of the Bose-Einstein condensation of the triplet states. PMID- 12721624 TI - The origin of multiple superconducting gaps in MgB2. AB - Magnesium diboride, MgB2, has the highest transition temperature (T(c) = 39 K) of the known metallic superconductors. Whether the anomalously high T(c) can be described within the conventional BCS (Bardeen-Cooper-Schrieffer) framework has been debated. The key to understanding superconductivity lies with the 'superconducting energy gap' associated with the formation of the superconducting pairs. Recently, the existence of two kinds of superconducting gaps in MgB2 has been suggested by several experiments; this is in contrast to both conventional and high-T(c) superconductors. A clear demonstration of two gaps has not yet been made because the previous experiments lacked the ability to resolve the momentum of the superconducting electrons. Here we report direct experimental evidence for the two-band superconductivity in MgB2, by separately observing the superconducting gaps of the sigma and pi bands (as well as a surface band). The gaps have distinctly different sizes, which unambiguously establishes MgB2 as a two-gap superconductor. PMID- 12721625 TI - Extreme crustal oxygen isotope signatures preserved in coesite in diamond. AB - The anomalously high and low oxygen isotope values observed in eclogite xenoliths from the upper mantle beneath cratons have been interpreted as indicating that the parent rock of the eclogites experienced alteration on the ancient sea floor. Recognition of this genetic lineage has provided the foundation for a model of the evolution of the continents whereby imbricated slabs of oceanic lithosphere underpin and promote stabilization of early cratons. Early crustal growth is thought to have been enhanced by the addition of slab-derived magmas, leaving an eclogite residuum in the upper mantle beneath the cratons. But the oxygen isotope anomalies observed in eclogite xenoliths are small relative to those in altered ocean-floor basalt and intermediate-stage subduction-zone eclogites, and this has hindered acceptance of the hypothesis that the eclogite xenoliths represent subducted and metamorphosed ocean-floor basalts. We present here the oxygen isotope composition of eclogitic mineral inclusions, analysed in situ in diamonds using an ion microprobe/secondary ion mass spectrometer. The oxygen isotope values of coesite (a polymorph of SiO2) inclusions are substantially higher than previously reported for xenoliths from the subcratonic mantle, but are typical of subduction-zone meta-basalts, and accordingly provide strong support for the link between altered ocean-floor basalts and mantle eclogite xenoliths. PMID- 12721626 TI - Migration of a Late Cretaceous fish. AB - Late Cretaceous sediments from the Western Interior of North America yield exceptionally well preserved fossils that serve as proxies for the rapidly changing climate preceding the Cretaceous/Tertiary boundary (about 67-65 Myr ago). Here we reconstruct the ontogenetic history of a Maastrichtian-age fish, Vorhisia vulpes, by using the carbon, oxygen and strontium isotope ratios of four aragonite otoliths collected from the Fox Hills Formation of South Dakota. Individuals of V. vulpes spawned in brackish water (about 70-80% seawater) and during their first year migrated to open marine waters of the Western Interior Seaway, where they remained for 3 years before returning to the estuary, presumably to spawn and die. The mean delta(18)O from the marine growth phase of V. vulpes yields a seawater temperature of 18 degrees C, which is consistent with leaf physiognomy and general-circulation-model temperature estimates for the Western Interior during the latest Maastrichtian. PMID- 12721627 TI - Fitness costs of R-gene-mediated resistance in Arabidopsis thaliana. AB - Resistance genes (R-genes) act as an immune system in plants by recognizing pathogens and inducing defensive pathways. Many R-gene loci are present in plant genomes, presumably reflecting the need to maintain a large repertoire of resistance alleles. These loci also often segregate for resistance and susceptibility alleles that natural selection has maintained as polymorphisms within a species for millions of years. Given the obvious advantage to an individual of being disease resistant, what prevents these resistance alleles from being driven to fixation by natural selection? A cost of resistance is one potential explanation; most models require a lower fitness of resistant individuals in the absence of pathogens for long-term persistence of susceptibility alleles. Here we test for the presence of a cost of resistance at the RPM1 locus of Arabidopsis thaliana. Results of a field experiment comparing the fitness of isogenic strains that differ in the presence or absence of RPM1 and its natural promoter reveal a large cost of RPM1, providing the first evidence that costs contribute to the maintenance of an ancient R-gene polymorphism. PMID- 12721628 TI - Non-classical receptive field mediates switch in a sensory neuron's frequency tuning. AB - Animals have developed stereotyped communication calls to which specific sensory neurons are well tuned. These communication calls must be discriminated from environmental signals such as those produced by prey. Sensory systems might have evolved neural circuitry to encode both categories. In weakly electric fish, prey and communication signals differ in their spatial extent and frequency content. Here we show that stimuli of different spatial extents mimicking prey and communication signals cause a switch in the frequency tuning and spike-timing precision of electrosensory pyramidal neurons, resulting in the selective and optimal encoding of both stimulus categories. As in other sensory systems, pyramidal neurons respond only to stimuli located within a restricted region of space known as the classical receptive field (CRF). In some systems, stimulation outside the CRF but within a non-classical receptive field (nCRF) can modulate the neural response to CRF stimulation even though nCRF stimulation alone fails to elicit responses. We show that pyramidal neurons possess a nCRF and that it can modulate the response to CRF stimuli to induce this neurobiological switch in frequency tuning. PMID- 12721630 TI - Genome sequence of Bacillus cereus and comparative analysis with Bacillus anthracis. AB - Bacillus cereus is an opportunistic pathogen causing food poisoning manifested by diarrhoeal or emetic syndromes. It is closely related to the animal and human pathogen Bacillus anthracis and the insect pathogen Bacillus thuringiensis, the former being used as a biological weapon and the latter as a pesticide. B. anthracis and B. thuringiensis are readily distinguished from B. cereus by the presence of plasmid-borne specific toxins (B. anthracis and B. thuringiensis) and capsule (B. anthracis). But phylogenetic studies based on the analysis of chromosomal genes bring controversial results, and it is unclear whether B. cereus, B. anthracis and B. thuringiensis are varieties of the same species or different species. Here we report the sequencing and analysis of the type strain B. cereus ATCC 14579. The complete genome sequence of B. cereus ATCC 14579 together with the gapped genome of B. anthracis A2012 enables us to perform comparative analysis, and hence to identify the genes that are conserved between B. cereus and B. anthracis, and the genes that are unique for each species. We use the former to clarify the phylogeny of the cereus group, and the latter to determine plasmid-independent species-specific markers. PMID- 12721632 TI - Seeking out the elite. PMID- 12721631 TI - An expressed pseudogene regulates the messenger-RNA stability of its homologous coding gene. AB - A pseudogene is a gene copy that does not produce a functional, full-length protein. The human genome is estimated to contain up to 20,000 pseudogenes. Although much effort has been devoted to understanding the function of pseudogenes, their biological roles remain largely unknown. Here we report the role of an expressed pseudogene-regulation of messenger-RNA stability-in a transgene-insertion mouse mutant exhibiting polycystic kidneys and bone deformity. The transgene was integrated into the vicinity of the expressing pseudogene of Makorin1, called Makorin1-p1. This insertion reduced transcription of Makorin1-p1, resulting in destabilization of Makorin1 mRNA in trans by way of a cis-acting RNA decay element within the 5' region of Makorin1 that is homologous between Makorin1 and Makorin1-p1. Either Makorin1 or Makorin1-p1 transgenes could rescue these phenotypes. Our findings demonstrate a specific regulatory role of an expressed pseudogene, and point to the functional significance of non-coding RNAs. PMID- 12721629 TI - The genome sequence of Bacillus anthracis Ames and comparison to closely related bacteria. AB - Bacillus anthracis is an endospore-forming bacterium that causes inhalational anthrax. Key virulence genes are found on plasmids (extra-chromosomal, circular, double-stranded DNA molecules) pXO1 (ref. 2) and pXO2 (ref. 3). To identify additional genes that might contribute to virulence, we analysed the complete sequence of the chromosome of B. anthracis Ames (about 5.23 megabases). We found several chromosomally encoded proteins that may contribute to pathogenicity- including haemolysins, phospholipases and iron acquisition functions--and identified numerous surface proteins that might be important targets for vaccines and drugs. Almost all these putative chromosomal virulence and surface proteins have homologues in Bacillus cereus, highlighting the similarity of B. anthracis to near-neighbours that are not associated with anthrax. By performing a comparative genome hybridization of 19 B. cereus and Bacillus thuringiensis strains against a B. anthracis DNA microarray, we confirmed the general similarity of chromosomal genes among this group of close relatives. However, we found that the gene sequences of pXO1 and pXO2 were more variable between strains, suggesting plasmid mobility in the group. The complete sequence of B. anthracis is a step towards a better understanding of anthrax pathogenesis. PMID- 12721633 TI - Getting organized. PMID- 12721634 TI - A genetic linkage map for hexaploid, cultivated oat (Avena sativa L.) based on an intraspecific cross 'Ogle/MAM17-5'. AB - Genetic research and breeding of oat ( Avena sativa L.) would be aided by development of a genetic linkage map for a breeding population. Such a map could be used for localization of qualitative and quantitative trait loci, marker assisted selection and other genetic analysis in an adapted, agronomically useful background. The objectives of this research were to develop a genetic linkage map of hexaploid cultivated oat, to identify homoeologous relationships of linkage groups, and to compare homologous linkage groups between this map and the previously published hexaploid oat map from the cross 'Kanota/Ogle' (KO). A total of 510 markers, including 172 restriction fragment length polymorphisms (RFLP), 324 amplified fragment length polymorphisms (AFLP) and 14 simple sequence repeats (SSR), were assessed on a recombinant inbred population of 152 F(5:6) lines derived from the cross, 'Ogle/MAM17-5' (OM). Twenty eight linkage groups of 5 cM or longer were formed using 476 of the markers, while 34 markers remained either unlinked or in small fragments less than 5 cM. The 28 linkage groups contained from 3 to 33 markers, and varied in size from 5.2 to 123.0 cM, representing a total map length of 1,396.7 cM. Three putative homoeologous groups (OM7, OM8 and OM18; OM2 and OM23; OM13 and OM16) were identified. Comparison with the published KO map indicated that nine OM linkage groups could be determined to be homologous to linkage groups in the KO map. Further comparison of the homologous linkage groups revealed that residual differences in genomic rearrangements existed between the two hexaploid oat populations. Some linkage groups were significantly extended compared with the KO map. Since the OM mapping population is segregating for a number of agronomically important traits, this genetic map will provide a useful tool for identification of qualitative and quantitative loci for these traits. PMID- 12721635 TI - Gene actions of QTLs affecting several agronomic traits resolved in a recombinant inbred rice population and two testcross populations. AB - To understand the types of gene action controlling seven quantitative traits in rice, QTL mapping was performed to dissect the main effect (M-QTLs) and digenic epistatic (E-QTLs) QTLs responsible for the trait performance of 254 recombinant inbred lines (RILs) of "Lemont/Teqing", and two testcross (TC) F(1) populations derived from these RILs. The correlation analyses reveal a general pattern, i.e. trait heritability in the RILs was negatively correlated to trait heterosis in the TC hybrids. A large number of M-QTLs and E-QTLs affecting seven traits, including heading date (HD), plant height (PH), flag leaf length (FLL), flag leaf width (FLW), panicle length (PL), spikelet number per panicle (SN) and spikelet fertility (SF), were identified and could be classified into two predominant groups, additive QTLs detected primarily in the RILs, and overdominant QTLs identified exclusively in the TC populations. There is little overlap between QTLs identified in the RILs and in the TC populations. This result implied that additive gene action is largely independent from non-additive gene action in the genetic control of quantitative traits of rice. The detected E-QTLs collectively explained a much greater portion of the total phenotypic variation than the M QTLs, supporting prior findings that epistasis has played an important role in the genetic control of quantitative traits in rice. The implications of these results to the development of inbred and hybrid cultivars were discussed. PMID- 12721636 TI - Leaf-rust resistance in rye (Secale cereale L.). 1. Genetic analysis and mapping of resistance genes Pr1 and Pr2. AB - Genetic analysis of resistance to leaf rust in rye (Puccinia recondita f. sp. secalis) led to the identification of two dominant resistance genes, Pr1 and Pr2. Both genes proved to be effective against a local leaf-rust population as well as a subset of single-pustule isolates (SPIs) the latter of which comprised SPIs with very high virulence complexity. Resistance conferred by Pr1 and Pr2 was expressed in detached-leaf tests of seedlings as well as in field tests of adult plants. Molecular marker analysis allowed us to map Pr1 in the proximal part of rye chromosome 6RL, whereas Pr2 was assigned to the distal part of chromosome 7RL. These results are discussed in view of homoeology relationships among Triticeae. A proposal is submitted for the designation of resistance genes to rye leaf rust which would avoid interference with existing gene-symboling in respect to wheat leaf-rust resistances introgressed from rye into wheat or triticale. PMID- 12721637 TI - Two critical factors are required for efficient transformation of multiple soybean cultivars: Agrobacterium strain and orientation of immature cotyledonary explant. AB - An efficient transformation system was developed for multiple soybean [Glycine max (L.) Merrill.] cultivars using Agrobacterium-mediated gene transfer. A significantly high number of hygromycin-resistant somatic embryos (SEs) was obtained when immature zygotic cotyledons were inoculated with Agrobacterium tumefaciens strain KYRT1 and when the abaxial side of explants was oriented upwards (i.e., the adaxial side of explants was in contact with the medium). Most hygromycin-resistant SEs on selective medium were induced along the periphery of the abaxial side of cotyledonary explants. Extended periods of selection (up to 10 weeks post-cocultivation) increased the frequency of somatic embryogenesis, and more than 50% of selected SEs tested positive for beta-glucuronidase (GUS). Following maturation and regeneration of selected SEs, ten independent transgenic soybean plants of cv Jack were obtained, and the overall transformation frequency ranged from 1.1 to 1.7%. Six and two transgenic plantlets were obtained from cvs Dwight and Williams, respectively. In addition, transgenic suspension lines were established from cvs Jack, Williams, Dwight, Rend and Ina. Molecular analysis of embryogenic lines and/or transgenic plants, established from different cultivars, confirmed stable integration, expression, and/or inheritance of transgenes in both T0 and T1 plants. PMID- 12721639 TI - Hybridization between transgenic Brassica napus L. and its wild relatives: Brassica rapa L., Raphanus raphanistrum L., Sinapis arvensis L., and Erucastrum gallicum (Willd.) O.E. Schulz. AB - The frequency of gene flow from Brassica napus L. (canola) to four wild relatives, Brassica rapa L., Raphanus raphanistrum L., Sinapis arvensis L. and Erucastrum gallicum (Willd.) O.E. Schulz, was assessed in greenhouse and/or field experiments, and actual rates measured in commercial fields in Canada. Various marker systems were used to detect hybrid individuals: herbicide resistance traits (HR), green fluorescent protein marker (GFP), species-specific amplified fragment length polymorphisms (AFLPs) and ploidy level. Hybridization between B. rapa and B. napus occurred in two field experiments (frequency approximately 7%) and in wild populations in commercial fields (approximately 13.6%). The higher frequency in commercial fields was most likely due to greater distance between B. rapa plants. All F(1) hybrids were morphologically similar to B. rapa, had B. napus- and B. rapa-specific AFLP markers and were triploid (AAC, 2n=29 chromosomes). They had reduced pollen viability (about 55%) and segregated for both self-incompatible and self-compatible individuals (the latter being a B. napus trait). In contrast, gene flow between R. raphanistrum and B. napus was very rare. A single R. raphanistrum x B. napus F1 hybrid was detected in 32,821 seedlings from the HR B. napus field experiment. The hybrid was morphologically similar to R. raphanistrum except for the presence of valves, a B. napus trait, in the distorted seed pods. It had a genomic structure consistent with the fusion of an unreduced gamete of R. raphanistrum and a reduced gamete of B. napus (RrRrAC, 2n=37), both B. napus- and R. raphanistrum-specific AFLP markers, and had <1% pollen viability. No hybrids were detected in the greenhouse experiments (1,534 seedlings), the GFP field experiment (4,059 seedlings) or in commercial fields in Quebec and Alberta (22,114 seedlings). No S. arvensis or E. gallicum x B. napus hybrids were detected (42,828 and 21,841 seedlings, respectively) from commercial fields in Saskatchewan. These findings suggest that the probability of gene flow from transgenic B. napus to R. raphanistrum, S. arvensis or E. gallicum is very low (<2-5 x 10(-5)). However, transgenes can disperse in the environment via wild B. rapa in eastern Canada and possibly via commercial B. rapa volunteers in western Canada. PMID- 12721640 TI - Linkage mapping of Hsa-1(Og), a resistance gene of African rice to the cyst nematode, Heterodera sacchari. AB - Inheritance of resistance to cyst nematode (Heterodera sacchari) in Oryza sativa was investigated by inoculation tests with isolate 244 from Congo in segregating populations derived from hybridisation between O. sativa and its African sister cultivated species, O. glaberrima. We found that the resistance was controlled by one major gene, Hsa-1(Og), with codominance of susceptible and resistant alleles. To map Hsa-1(Og) on the rice genome, we pooled the data obtained from segregation of the resistance trait and microsatellite markers in three kinds of progeny: BC(1)F(3), BC(1)F(4), and pseudo-F(2) populations. Hsa-1(Og) was unambiguously located between Cornell University's RM206 and RM254 markers on chromosome 11. Two additional microsatellite markers derived from Monsanto publicly available sequences were found to be tightly linked to the Hsa-1(Og) gene. It is possible that numerous plant resistances to a pathogen in fact exhibit a codominant inheritance, possibly explaining misleading conclusions in several reports on resistance segregation. PMID- 12721642 TI - [Primary hyperparathyroidism, adrenal tumors and neuroendocrine tumors of the pancreas--clinical diagnosis and imaging requirements]. AB - INTRODUCTION: Diseases of the parathyroids, the adrenals and of neuroendocrine tumors of the pancreas are primarily diagnosed by clinical and endocrinological evaluation. The requirements concerning various imaging techniques and their relative importance in localization strategies of the different tumors are complex. MATERIALS AND METHODS: Current literature search, using PubMed. RESULTS: Evaluation of primary hyperparathyroidism requires bone densitometry by DXA and search for nephrolithiasis by ultrasound or native CT examination. While ultrasound of the thyroid and parathyroids seems useful before any parathyroid surgery, more extensive preoperative localization strategies (sestamibi scintigraphy, MRI) should be restricted to minimal invasive parathyroid surgery or reoperations. For adrenal tumors CT and MRI are of similar diagnostic value. Imaging of pheochromocytomas should be completed by MIBG scintigraphy. Each adrenal incidentaloma requires an endocrinological work-up. A fine-needle aspiration or core needle biopsy of an adrenal tumor is rarely indicated. Before adrenal biopsy a pheochromocytoma has to be excluded. Successful localization strategies for neuroendocrine tumors of the pancreas include somatostatin receptor scintigraphy, endoscopic ultrasound and MRI. DISCUSSION: Specific localization strategies have been established for the aforementioned tumors. The continuous progress of different imaging techniques requires a regular reevaluation of these localization strategies. PMID- 12721643 TI - [Diagnosis of hyperparathyroidism]. AB - AIM: To preoperatively localize enlarged parathyroid glands in patients with hyperparathyroidism (HPT). METHODS: Besides clinical and biochemical workup, high resolution ultrasonography (US) is the most commonly used imaging method.Additionally,Tc-99m-MIBI scintigraphy in subtraction or biphasic technique, computed tomography (CT) and magnetic resonance imaging (MRI) are used. RESULTS: US fails to detect a minimum of 10% of enlarged parathyroid glands,most commonly due to ectopic location, or difficult examination conditions, such as nodular goiter or previous surgery. If attempted US localization is unsuccessful, multiphase scintigraphy, using Tc-99m-sestamibi, can help to locate ectopic adenomas in the mediastinum. With SPECT, ademomas can be found which escape detection on planar scans due to their small size. With combined use of US and scintigraphy, a correct localization of parathyroid adenomas is possible in up to 90%. CT and MRI are of limited value due to their low specificity. CONCLUSION: For newly diagnosed hyperparathyroidism, high resolution US is the method of choice for localizing parathyroid adenomas. If ultrasound fails to detect a lesion, Tc-99m-MIBI scintigraphy is recommended. In patients scheduled for re-operation for recurrent or persistent HPT, a preoperative detection of a parathyroid adenoma should be attempted whenever possible, in order to minimize the extent of surgery. The role of CT or MRI is mainly to help to better anatomically localize a suspected adenoma previously detected with scintigraphy. PMID- 12721644 TI - [Value of various imaging modalities for diagnostic work-up of tumors of the adrenal gland]. AB - OBJECTIVE: This paper describes the value of various imaging modalities for diagnostic work-up of tumors of the adrenal gland. METHODS: Results of the literature are reviewed. An optimized examination protocol for computed tomography (CT) and magnetic resonance imaging (MRI) is shown for assessment and differentiation of unclear lesions of the adrenal gland. RESULTS: Measurements of attenuation in the native examination as well as delayed enhancement are the cornerstones in the CT diagnostics of tumors of the adrenal gland. In MRI, chemical-shift imaging and evaluation of signal characteristics in T1- and T2 weighted images are most important for characterization even in unclear cases. CONCLUSION: CT and MRI play the major role in imaging of adrenal gland tumors. Whereas CT is less expensive and widely available,MRI provides advantages in unclear cases because of the excellent tissue contrast and the superior characterization. PMID- 12721645 TI - [Imaging of neuroendocrine tumors of the pancreas]. AB - OBJECTIVE: To compare the value of the various imaging modalities in the detection of pancreatic islet cell tumors. METHODS: Analysis of the literature to compare the value of ultrasound, CT, MRI, angiography, venous sampling, scintigraphy and PET in the detection of neuroendocrine tumors of the pancreas. RESULTS: For transabdominal ultrasound a sensitivity of 60% has been reported; for endoscopic ultrasound a sensitivity of more than 90%. MRI with a sensitivity of 75-100% is more sensitive compared to CT with a sensitivity of 64-82%. Angiography and venous sampling have a reported sensitivity of 66% and 76%. Somatostatin receptor scintigraphy has a sensitivity of 81-88% with significant differences between the various histological subtypes of the pancreatic islet cell tumors. PET does not play an important role in the detection of neuroendocrine tumors of the pancreas. CONCLUSION: None of the various modalities is superior alone, so that it is often necessary to combine them for diagnostic imaging of pancreatic islet cell tumors. PMID- 12721647 TI - [Experience with a quality control system for mammography in a private practice setting]. AB - Design and performance of a private audit of mammography utilizing the BI-RADS classification in a private practice setting are presented. By using a combination of physical examination,mammography and, whenever necessary, additional views and sonography, the goals of the BI-RADS audit were met. Practical issues of this audit are addressed. PMID- 12721646 TI - [Intraarterial calcium stimulation (ASVS) for pancreatic insulinoma: comparison of preoperative localization procedures]. AB - PURPOSE: Evaluation of clinical relevance of the arterial stimulation procedure with venous sampling (ASVS) in the preoperative localization of insulinoma. METHODS: Thirteen patients with endogenous hyperinsulinism underwent preoperative transabdominal ultrasound (US), helical CT (CT), MRI, endoscopic ultrasound (EUS), and angiography (DSA) in conjunction with the ASVS-test for the detection of insulinoma. The results were compared with intraoperative findings, intraoperative ultrasound (IOUS) and histology. RESULTS: Sensitivity was as follows: US 8%, MRI 27%, CT 46%, EUS 50%,DSA 69%,and ASVS 92%. Intraoperative palpation and IOUS yielded a sensitivity of 77%. In 3 patients the tumors were neither palpable nor detectable by IOUS, the mode of resection was based on preoperative diagnostics. The ASVS procedure as a functional test was superior to all other modalities for the preoperative tumor detection. CONCLUSION: The ASVS was the most sensitive diagnostic modality. It should especially be considered in terms of health economical aspects when CT or MRI do not yield conclusive results. PMID- 12721648 TI - [Application of multislice detector spiral computed tomography to intracranial aneurysms: first clinical experience]. AB - PURPOSE: To identify advantages when using the new multislice spiral CT technology (MSCT) for the detection and demonstration of intracranial aneurysms with CT angiography (CTA)? MATERIAL AND METHODS: Patients with suspected intracranial aneurysms underwent 4- and 8-slice spiral CTA. Image postprocessing included "maximum intensity projection" (MIP),"volume rendering" (VR) and additionally curved multiplanar reconstructions (MPR) for aneurysms suspected at the carotid siphon. The results were compared with those achieved with digital subtraction angiography (DSA). RESULTS: In 19 patients 21 out of 22 aneurysms were detected by CTA. A high resolution three-dimensional view of the aneurysm morphology in spatial relation to the surrounding structures was achieved. Further information could be gained by making a detailed examination of the inside wall of the aneurysm, using virtual vascular 3D-endoscopy and virtual 3D craniotomy. CONCLUSION: Compared to DSA,MS-CTA is a useful additional and in selected cases alternative modality for the detection with marked advantages in the demonstration of intracranial aneurysms. PMID- 12721649 TI - [Inflammatory diseases of the orbit. Part 1: Intraconal space]. PMID- 12721663 TI - Cloning and characterization of a novel cardiac-specific kinase that interacts specifically with cardiac troponin I. AB - Cardiac-restricted genes play important roles in cardiovascular system. In an effort to identify such novel genes we identified a novel cardiac-specific kinase gene TNNI3K localized on 1p31.1 based on bioinformatics analyses. Sequence analysis suggested that TNNI3K is a distant family member of integrin-linked kinase. Northern blot and 76-tissue array analyses showed that TNNI3K is highly expressed in heart, but is undetectable in other tissues. Immunohistochemical analysis predominantly localized TNNI3K in the nucleus of cardiac myocytes. In vitro kinase assay showed that TNNI3K is a functional kinase. The yeast two hybrid system showed that TNNI3K could directly interact with cardiac troponin I, results that were further confirmed by coimmunoprecipitation in vivo. Our data suggest that TNNI3K is a cardiac-specific kinase and play important roles in cardiac system. PMID- 12721664 TI - B7-H1 pathway and its role in the evasion of tumor immunity. AB - B7-H1 is a recently identified member of the B7 family molecules. Upon ligation to its receptors on T cells it regulates activation and differentiation of T cells. B7-H1 preferentially costimulates IL-10 production in resting T cells and further induces the apoptosis of activated T cells. PD-1 is a receptor of B7-H1 and is shown to mediate the inhibition of activated T cell response, presumably by inhibiting cell cycle progression. The expression of B7-H1 protein is limited to macrophage lineage of cells in normal tissues, although its mRNA transcription is found in a broad range of tissues. In contrast, B7-H1 is abundant in various human cancers. The tumor-associated B7-H1 increases apoptosis of antigen specific T cells, leading to growth of immunogenic tumor growth in vivo. Current data suggest that B7-H1 regulates the organ-specific tolerance in normal tissue and may contribute to immune evasion by cancers. Selective manipulation of B7-H1 pathway thus aids in the design of new regimens in the treatment of human autoimmune disease and the control of malignant cancers. PMID- 12721665 TI - Lentivirus-mediated gene transfer of uroporphyrinogen III synthase fully corrects the porphyric phenotype in human cells. AB - Congenital erythropoietic porphyria (CEP) is an inherited disease due to a deficiency in the uroporphyrinogen III synthase, the fourth enzyme of the heme biosynthesis pathway. It is characterized by accumulation of uroporphyrin I in the bone marrow, peripheral blood and other organs. The prognosis of CEP is poor, with death often occurring early in adult life. For severe transfusion-dependent cases, when allogeneic cell transplantation cannot be performed, the autografting of genetically modified primitive/stem cells may be the only alternative. In vitro gene transfer experiments have documented the feasibility of gene therapy via hematopoietic cells to treat this disease. In the present study lentiviral transduction of porphyric cell lines and primary CD34(+) cells with the therapeutic human uroporphyrinogen III synthase (UROS) cDNA resulted in both enzymatic and metabolic correction, as demonstrated by the increase in UROS activity and the suppression of porphyrin accumulation in transduced cells. Very high gene transfer efficiency (up to 90%) was achieved in both cell lines and CD34(+) cells without any selection. Expression of the transgene remained stable over long-term liquid culture. Furthermore, gene expression was maintained during in vitro erythroid differentiation of CD34(+) cells. Therefore the use of lentiviral vectors is promising for the future treatment of CEP patients by gene therapy. PMID- 12721667 TI - [Two consecutive pregnancies in a type 1 diabetic under treatment with insulin Lispro]. AB - BACKGROUND: Glucose control is mandatory in pregnant women with type 1 diabetes to keep the incidence of malformations and complications low. Insulin Lispro is a new and popular fast-acting insulin analog used for intensified insulin therapy. Although popular among young patients, this drug has not been licensed for use in pregnancy due to a possibly increased risk of fetal malformations. CASE REPORT: Retrospectively, the case of a 34-year-old woman with diabetes type 1, gravid III and para III, which carried her second and third child full time under Lispro treatment, was analyzed. We performed an interview of the patient and her doctors. Additionally, the pregnancy documents along with the examination documents of the children, our patient files, and the blood glucose documentation were investigated. Contrary to the first pregnancy which was carried out in the absence of Lispro (first degree hip dysplasia), both subsequent pregnancies were completely normal despite continuous Lispro therapy. HbA(1c) values in the range of 4.9-5.9% showed an adequate glucose control throughout these pregnancies. CONCLUSION: This case supports the hypothesis that insulin Lispro which is highly popular among young patients can safely and effectively be used in pregnancies. In order to be able to adequately assess possible risks associated with this application, additional cases need to be documented and analyzed. PMID- 12721666 TI - [The selective cyclooxygenase-2 inhibitor celecoxib is a safe alternative in patients with pseudo-allergic reactions to nonsteroidal anti-inflammatory drugs]. AB - BACKGROUND: Adverse reactions to nonsteroidal anti-inflammatory drugs (NSAIDs) are frequent, and the need to identify a safe alternative drug is a common problem in clinical practice. We assessed the tolerability of the selective cyclooxygenase-2 inhibitor celecoxib in a group of NSAID-sensitive patients. PATIENTS AND METHODS: 77 patients (24 males, 53 females, age 31-80 years) with a history of adverse reactions to NSAIDs underwent standardized skin prick, scratch and patch tests along with oral, placebo-controlled blinded exposure to celecoxib (maximum single dose 200 mg, cumulative daily dose 350 mg). RESULTS: 21 patients had a history of cutaneous reactions only (urticaria), 25 had encountered respiratory symptoms (asthma), 18 reported cutaneous as well as respiratory symptoms, and in 13 patients an anaphylactoid shock occurred. Acetylsalicylic acid triggered symptoms in 38 patients. In 46 cases, several NSAIDs of different chemical groups caused symptoms. Oral challenge with celecoxib was tolerated by all 77 patients without adverse effects. CONCLUSION: This study demonstrates that celecoxib does not have cross-intolerance to NSAIDs. Celecoxib is a safe alternative in subjects with previous adverse reactions to NSAIDs. PMID- 12721668 TI - [Apoptosis induced by free fatty acids]. AB - BACKGROUND AND AIMS: Elevated free fatty acid (FFA) levels are believed to be one of the major contributing factors in the pathogenesis of type 2 diabetes. FFAs enhance peripheral insulin resistance, promote beta-cell dysfunction, and trigger beta-cell death. The purpose of this study was to investigate whether the lipoapoptotic effect of FFAs is determined by the degree of saturation and the chain length, and which pathways might be involved. MATERIAL AND METHODS: After incubation of RIN1046-38 rat insulinoma cells and primary human islet cells with different FFAs, apoptosis was assessed by flow cytometry (sub-G1-DNA formation) and TUNEL assay, respectively. Intracellular distribution of protein kinase C delta (PKC-delta) was determined by confocal laser microscopy. RESULTS: In RIN1046-38 cells, the sub-G1 fraction increased 40-fold over basal (0.5% to approximately 20%) after treatment with the saturated FFAs palmitate and stearate (1 mM, 24 h), while the monounsaturated FFAs palmitoleate and oleate as well as the polyunsaturated FFA linoleate had no proapoptotic effect. Unsaturated FFAs even more prevented palmitate- and stearate-induced apoptosis completely. Furthermore, these results could be confirmed in primary beta-cells of human islets. Investigating potential underlying signaling pathways, it was found that only the saturated FFAs were able to induce nuclear translocation of PKC-delta. CONCLUSIONS: FFAs exert different effects on beta-cell survival. Saturated FFAs turned out to be pro-apoptotic, whereas unsaturated FFAs protected from saturated FFA-induced apoptosis. Furthermore, our data suggest an involvement of PKC-delta in apoptosis induced by saturated FFAs. PMID- 12721669 TI - [Urinary albumin excretion and noninvasive assessment of peripheral endothelial dysfunction with high-resolution ultrasound in type 2 diabetic subjects and nondiabetic controls]. AB - BACKGROUND: Recent theory in pathogenesis of atherosclerosis has focused on the pathobiology of the artery wall including the emerging influence of the nitric oxide (NO) system on thrombogenicity and trigger mechanisms leading to morphologic changes culminating in the stenotic plaque. Therefore, diagnostic evaluation of disturbances in NO bioavailability might be of prognostic relevance regarding primary prevention of cardiovascular disease. Disturbances in NO production can be measured noninvasively with conventional high-resolution ultrasound. On the other hand, particularly in individuals with diabetes, microalbuminuria is thought to be associated with an increased risk of cardiovascular events. Thereby it is still unknown, whether an increase in renal albumin excretion can be regarded as an indicator of global endothelial dysfunction, or whether other partial functions such as the nitric oxide system might be disturbed earlier. PROBANDS AND METHODS: Therefore, the NO system and renal albumin excretion were examined in 129 subjects (56 with type 2 diabetes and 73 nondiabetics). Nitric oxide production was assessed by measuring flow mediated vasodilatation (FMD) of the brachial artery using a 13-MHz linear array. Comparison was done between subjects with disturbed endothelial NO production (FMD < 5%) and subjects with normal regulation of the vascular tone (FMD > 5%). RESULTS: In normoalbuminuric individuals (< 20 microg/min, and < 20 mg/l, respectively), neither for the group of subjects with type 2 diabetes nor in the group of nondiabetics, relevant differences could be found in renal albumin excretion (RAE) rate between subjects with disturbed and normal FMD (RAE in diabetics 4.8 +/- 5.5 vs. 4.6 +/- 5.1 mg/l and in nondiabetics 5.1 +/- 2.6 vs. 4.9 +/- 2.7 microg/min). Both groups were well balanced regarding other risk factors of the metabolic syndrome (systolic/diastolic blood pressure, glucose and lipid metabolism). Furthermore, comparison of FMD in subjects with microalbuminuria (20-200 microg/min and 20-200 mg/l, respectively, n = 18) versus normoalbuminuric individuals (n = 111) again did not reveal a significant difference for the diabetic group (FMD median 4.3% [range 1.8-7.6%] vs. 5.0% [range 1.1-9.1%]) nor for the nondiabetic group (FMD median 4.7% [range 3.1 13.3%] vs. 5.2% [range -1.2-31.6%]). However, this analysis underlined the considerable influence of the classic cardiovascular risk factors. Particularly in the nondiabetic group, individuals with microalbuminuria showed higher blood pressure (p = 0.05) and a higher body mass index (p < 0.01). CONCLUSION: From these results, it is concluded that both procedures (renal albumin excretion rate and the measurement of endothelium-dependent vasodilatation) investigate two independent disturbances of the vascular wall. Furthermore, these results lead to the hypothesis that disturbances in endothelial NO production occur early and may already be operative before renal albumin excretion increases. Thus, for the purpose of actually identifying cardiovascular high-risk subjects early, peripheral endothelial dysfunction should be measured in addition to renal albumin excretion rate. PMID- 12721670 TI - [Microbiological aspects and antibiotic therapy of diabetic foot infections]. AB - IMMUNOLOGICAL AND MICROBIOLOGICAL ASPECTS OF DIABETIC FOOT INFECTIONS: Diabetic patients are at increased risk of severe skin and bone infections. Immunological disturbances are reasonable and due to altered specific and unspecific cellular immune responses. Analysis of epidemiology and microbial pathogenicity shows that staphylococci seem to be predestined to induce such infections. Staphylococcus aureus and coagulase-negative staphylococci are able to adhere to the wound ground by a sequela of mechanisms. Initial bacterial adherence is due to hydrophobicity, ion exchanges, and specific binding of bacterial adhesion molecules to cellular receptors. Moreover, staphylococci secrete polysaccharides which form a biofilm together with multilayer cell clusters. The highly structured communities within a biofilm are resistant to distinct immunoeffectors and have a decreased susceptibility to antibiotics in vivo. ASPECTS OF ANTIBIOTIC THERAPY: Assessing the severity of an infection is essential to selecting an antibiotic regimen, the mode of drug administration, and the duration of therapy. Regimens for severe and chronic infections are broader spectrum and often intravenously to obtain high drug concentrations immediately. Infections of the bone often require an antibiotic therapy for > 4 weeks, while a 1- to 2-week therapy for mild to moderate infections has been found to be effective. CONCLUSIONS: Because of the tremendous progress in diagnostics and therapy of diabetic foot infections, infectious complications can be successfully treated by appropriate wound care, metabolic control, and early surgical and antibiotic intervention. Bacterial biofilms involved into chronic infections are new aspects currently not visualized by clinical therapy. Besides the classic antimicrobial therapy, new concepts of an enzymatic therapy or the inhibition of bacterial "communication" (quorum sensing) are in progress and the hope for the future. PMID- 12721671 TI - [Glucocorticoid-induced insulin resistance and diabetes mellitus. Receptor-, postreceptor mechanisms, local cortisol action, and new aspects of antidiabetic therapy]. AB - BACKGROUND: Glucocorticoids are frequently prescribed drugs. Nearly half of the patients treated with glucocorticoids over a longer period develop a deranged glucose metabolism. In about 50%, these disturbances persist despite reduction or even withdrawal of the drug. PATHOPHYSIOLOGY: Glucocorticoids antagonize the insulin-mediated inhibition of hepatic glucose release, decrease glucose utilisation in muscle, and reduce the binding affinity of insulin receptors. Therefore, glucocorticoid-induced diabetes mellitus is equivalent to unmasked type 2 diabetes. New studies presume that an increased endogenous production of glucocorticoids particularly in adipocytes could play a role in type 2 diabetes as well. THERAPY: Patients with glucocorticoid-induced diabetes bear, comparable to patients with other types of diabetes, a considerable risk of arteriosclerotic and cardiovascular diseases and should therefore receive an intensified treatment. Therapy of glucocorticoid-induced diabetes basically corresponds to that of type 2 diabetes. Applicable are oral antidiabetic drugs, particularly metformin and the glitazones as insulin sensitizers both requiring consideration of contraindications, or treatment with insulin. PMID- 12721672 TI - [The HIV protease inhibitor-induced insulin resistance syndrome]. AB - BACKGROUND: HIV protease inhibitors improve the morbidity and mortality of HIV infection, however, this therapy also initiates insulin resistance that is associated with an increasing development of disturbances in glucose and lipid metabolism. Similar to the common insulin resistance syndrome, treated patients also suffer from an increased cardiovascular risk. PATHOGENESIS: Studies based on animal or cell models suggest that HIV protease inhibitors induce short-term effects on GLUT4 glucose transport and longterm effects on insulin signal transduction. CONCLUSION: Therapy with HIV protease inhibitors should include regular monitoring of metabolic alterations and cardiovascular damage. Particularly patients with genetic or traditional risk factors for type 2 diabetes should be monitored frequently. PMID- 12721673 TI - [Current status and perspectives of stem cell therapy for the treatment of diabetes mellitus]. AB - Due to autoimmune destruction of insulin-producing pancreatic beta-cells, type 1 diabetic patients, and also patients with type 2 diabetes suffering from defective insulin secretion rely on lifelong substitution with insulin. A clinically established alternative therapy for diabetics with exogenous insulin substitution, the transplantation of human islets of Langerhans, is limited by the lack of donor organs. The intensive search for new sources of pancreatic beta cells now focuses on human stem cells. Insulin-producing cells for transplantation can be generated from both embryonic and adult pancreatic stem cells. Both types of stem cells, however, differ with respect to availability, in vitro expansion, potential for differentiation, and tumorigenicity, which is elucidated by the authors. Before stem cell therapeutic strategies for diabetes mellitus can be transferred to clinical application in humans, aspects of functional effectiveness, safety, and cost-effectiveness have to be solved. Considering these prerequisites in the light of currently available therapeutic options, however, it can be estimated, that stem cell therapy for diabetes mellitus may be cost-effectively introduced into clinical routine in the future. PMID- 12721674 TI - [Novel aspects in the mechanisms of steroid diabetes and the regulation of hepatic glucose production by insulin and steroids]. AB - Glucocorticoids are known to increase blood glucose levels, and an impairment of glucose tolerance is a common side effect of glucocorticoid therapy and a central feature of Cushing's disease. A major pathophysiological event in this process is an increased glucose production of the liver on the basis of glucocorticoid induced insulin resistance resulting in an increment in hepatic gluconeogenesis. Both, glucocorticoids and insulin are known to affect the expression of the two gluconeogenic key enzymes, phosphoenolpyruvate-carboxykinase (PEPCK) and glucose 6-phosphatase (G6Pase). While glucocorticoids are known to stimulate the expression of the PEPCK- and G6Pase gene, insulin decreases hepatic glucose production through an inhibition of PEPCK- and G6Pase gene expression. Recently, considerable progress has been made in the understanding of the signal transduction involved in the glucocorticoid- and insulin-dependent regulation of hepatic gluconeogenesis. In this article, we will review the most recent advances and assemble the current knowledge into a clinically relevant pathophysiological model. PMID- 12721675 TI - [Insulin secretion and aging. Implications for insulin therapy in the geriatric patient with diabetes mellitus]. AB - Diabetes mellitus has become a global epidemic. With diabetes prevalence highest in old age groups and aging populations, it is the prevalence among the elderly, above all, where most dramatic increases can be expected. The present paper summarizes the age-related metabolic abnormalities resulting in diabetes mellitus, specifically the progressive decline in beta-cell function. In addition, insulin therapy in the elderly is described. Among the many different insulins and therapeutic algorithms, advantages and disadvantages pertaining to the elderly will be highlighted. Specifics in frequency, pathophysiology, and avoidance of severe hypoglycemia in the elderly are discussed. PMID- 12721676 TI - [Molecular fundamentals of wound healing in diabetic foot syndrome]. AB - BACKGROUND: The diabetic foot syndrome represents a considerable problem of health care. Due to the systemic character of diabetes mellitus, disturbances on the molecular level of wound healing are assumed. PATHOGENESIS: Growth factors and proteases affect the process of normal wound healing, and changes of their activity are relevant to the pathogenesis of the chronic wound. First studies describe the transition of a diabetic foot lesion to a nonhealing chronic wound on the molecular and cellular level. CURRENT RESEARCH: This review reports on the current status of research and new implications for the treatment of diabetic foot syndrome. PMID- 12721677 TI - Bioinformatic and molecular analysis of hydroxymethylbutenyl diphosphate synthase (GCPE) gene expression during carotenoid accumulation in ripening tomato fruit. AB - Carotenoids are plastidic isoprenoid pigments of great biological and biotechnological interest. The precursors for carotenoid production are synthesized through the recently elucidated methylerythritol phosphate (MEP) pathway. Here we have identified a tomato ( Lycopersicon esculentum Mill.) cDNA sequence encoding a full-length protein with homology to the MEP pathway enzyme hydroxymethylbutenyl 4-diphosphate synthase (HDS, also called GCPE). Comparison with other plant and bacterial HDS sequences showed that the plant enzymes contain a plastid-targeting N-terminal sequence and two highly conserved plant specific domains in the mature protein with no homology to any other sequence in the databases. The ubiquitous distribution of HDS-encoding expressed sequence tags (ESTs) in the tomato collections suggests that the corresponding gene is likely expressed throughout the plant. The role of HDS in controlling the supply of precursors for carotenoid biosynthesis was estimated from the bioinformatic and molecular analysis of transcript abundance in different stages of fruit development. No significant changes in HDS gene expression were deduced from the statistical analysis of EST distribution during fruit ripening, when an active MEP pathway is required to support a massive accumulation of carotenoids. RNA blot experiments confirmed that similar transcript levels were present in both the wild-type and carotenoid-depleted yellow ripe ( r) mutant fruit independent of the stage of development and the carotenoid composition of the fruit. Together, our results are consistent with a non-limiting role for HDS in carotenoid biosynthesis during tomato fruit ripening. PMID- 12721678 TI - Nutrient translocation in the xylem of poplar--diurnal variations and spatial distribution along the shoot axis. AB - This investigation shows diurnal variations in the xylem sap composition of poplar (Populus tremula x P. alba). All major macronutrients reached a maximum concentration in the first half of the light period and decreased to the middle of the night. The relative abundance of the nutrients did not change during the day. The sap flow, which responded very fast to the environmental changes (2.2 fold increase within 10-20 min of illumination), reached a maximum value in the second half of the light period. Transpiration (and photosynthesis) was constant throughout the light phase. The calculated translocation rates displayed a maximum in the first half of the light period and therefore did not fit the time course of sap flow. During the night, translocation rates were 63-69% lower than the maximum. The regulation of nutrient translocation is discussed taking the active xylem loading into account. The axial distribution located the nitrate assimilation in younger leaves and storage of nitrate (and other macronutrients) in older leaves. Nitrate and potassium concentrations in the xylem sap did not change along the plant axis. However, the sap flow was greater in younger shoot sections than in older sections. We assume that the greater demand for nitrate in the younger shoot section was satisfied via an increased volume flow rather an increased nitrate concentration. PMID- 12721680 TI - Inflammatory bowel disease: management issues during pregnancy. AB - INTRODUCTION: Inflammatory bowel disease often affects women during their reproductive years, causing management concerns for obstetricians caring for these patients during pregnancy. TREATMENT: Apart from methotrexate, most drugs used regularly to treat ulcerative colitis and Crohn's disease can safely be used by pregnant women. No causal relationship has been established between exposure to sulfasalazine or other 5-aminosalicylic acid drugs and the development of congenital malformations and these drugs may be used with relative safety during pregnancy and lactation. Current evidence indicates that maternal use of azathioprine and mercaptopurine is not associated with an increased risk of congenital malformations, though impaired foetal immunity, intrauterine growth retardation and prematurity are occasionally observed. Cyclosporin is not teratogenic, but may be associated with growth retardation and prematurity. CONCLUSIONS: Pregnancy should be avoided in women treated with methotrexate because of its known abortifacient effects and risk of causing typical malformations. There is no actual evidence of adverse effect in pregnant women receiving Infliximab but the amount of clinical information is small. The treatment with metronidazole or ciprofloxacin for short durations appear to be safe, but there is no data about the effects of increased length of treatment as required in Crohn's disease remains unknown. Control of disease activity before conception and during pregnancy is critical to optimise both maternal and foetal health. A multidisciplined approach involving both obstetrician and gastroenterologist and education about pregnancy are essential components of the treatment of any young women with IBD. PMID- 12721682 TI - Halo jacket in odontoid fractures type II and III. AB - The treatment of odontoid fractures remains controversial. The late results of 14 patients with Anderson D'Alonzo type II and III treated with halo jacket from 1995 to 1999 are presented. The fractures were reduced under image intensifier and stabilized by halo jacket. The immobilization period was 12 weeks. After 1 year, the fracture was roentgenologically consolidated in 12 out of 14 patients (85.7%), and all the patients were without neurological deficit. Nine patients (64.3%) were without subjective complaints, five had a reduced range of motion. Painful motion appeared in four patients (28.6%), and cervical stiffness was noted in five (35.7%). We recommend halo jacket for the treatment of most type II and III odontoid fractures which are unstable or displaced 6 mm and more, except for patients with tetraplegia. Fractures with minimal displacement and without neurological deficit were treated conservatively with rigid collars. In our institution, internal fixation was performed in selected cases. The presence of neurological deficit and the anatomic properties of the fracture site crucially influence the choice of treatment. PMID- 12721681 TI - Reduced hepatic transcription factor activation and expression of IL-6 and ICAM-1 after hemorrhage by NO scavenging. AB - BACKGROUND: Hemorrhagic shock (HS) elicits an inflammatory response characterized by increased cytokine production and recruitment of polymorphonucleated neutrophilic granulocytes (PMN) that we reported to be inducible nitric oxide synthase (iNOS) dependent. In a previous study, we demonstrated that removing excess induced nitric oxide (NO) by administration of the NO scavenger NOX resulted in reduced PMN infiltration, attenuated liver injury, and improved survival. In this study, we examined the role of NOX treatment in down-modulating the inflammatory response in the liver following HS. METHODS: Rats ( n=5) were subjected to severe HS with mean arterial blood pressure (MAP) of 40 mmHg for 100 min followed by resuscitation and killing at 24 h. RESULTS: Shock animals demonstrated increased mRNA levels of interleukin (IL)-6 and intercellular adhesion molecule (ICAM)-1 and increased activation of the transcription factors nuclear factor kappa B (NF-kappa B) and signal transducers and activators of transcription 3 (Stat3). Treatment with NOX (30 mg/kg/h) infused 60 min following the onset of shock over 4 h resulted in significant reduction in cytokine mRNA expression and transcriptional factor activation. These results suggest that excessive NO contributes to hemorrhage-induced tissue inflammation and that reducing the bioavailability of NO using NOX may be beneficial in HS. CONCLUSION: These data indicate that NOX prevents liver injury in this HS model, possibly through down-modulation of proinflammatory signaling and the shock-induced inflammatory response. PMID- 12721679 TI - Myenteric nitrergic neurons along the rat esophagus: evidence for regional and strain differences in age-related changes. AB - Several studies have suggested an age-related reduction in the number of myenteric neurons in the lower gastrointestinal (GI) tract linked to changes in GI neuromuscular functions. The present study, combining protein gene product 9.5 immunostaining and NADPH-diaphorase histochemistry, aimed at quantifying the proportion of nitrergic neurons compared to the overall number of enteric neurons in the esophagus of young (3-4.5 months) and aged (18-20 months) Sprague-Dawley and Wistar rats. In both strains, the neuron numbers per ganglion in the cervical region were almost twice as high as in the other esophageal regions. Irrespective of age or strain, the esophagus harbored a very high proportion of intrinsic nitrergic neurons (greater than approximately 65%). Both strains showed with aging an overall neuronal loss of approximately 27%. While a significant increase (young: 64-71%; aged: 82-89%) was observed in all esophageal regions in the Wistar strain, the proportion of nitrergic neurons remained stable with aging in the Sprague-Dawley strain (range: 72-82%). In conclusion, the age-related reduction in the overall number of myenteric, nitrergic, and non-nitrergic neurons observed in the rat esophagus, appears to be highly region- and strain dependent. Therefore, a protective mechanism against neuronal cell loss, selectively present in specific (nitrergic) enteric subpopulations, as suggested in earlier reports, cannot be put forward as a general phenomenon throughout the entire GI tract. PMID- 12721683 TI - Age at onset, extent of necrosis, and containment in Perthes disease. Results at maturity. AB - BACKGROUND: The concept of containment of the femoral head within the acetabulum has been popularised as a principle of treatment in Perthes disease. The results of 30 patients treated for Perthes disease following a predefined scheme based on the containment were analyzed. METHODS: All children were treated with abduction braces as long as containment was maintained. However, 15 of them required surgery due to failure of containment in the further course of the disease. All children were followed until skeletal maturity. RESULTS: Within a mean follow-up period of 8 years and 9 months (range 5-13 years), all patients showed good clinical function. On radiographic assessment, 70% of the hips were Stulberg classes I and II, with a good prognosis. CONCLUSION: Failure of the containment was influenced by age at onset and extent of necrosis. Weight-bearing abduction braces cannot be recommended in children who are 6 years and older and show femoral head involvement of more than 50%. PMID- 12721684 TI - Proximal humeral fractures: how stiff should an implant be? A comparative mechanical study with new implants in human specimens. AB - BACKGROUND: The objective of this study was to determine the in vitro characteristics of the clinically used and newly developed implants for the stabilization of proximal humeral fractures under static and cyclic loading. The goal was to optimize implant stiffness for fracture stabilization even in weak bone stock. METHODS: In a laboratory study using 35 fresh human humeri, the specimens were randomized into 5 groups, which included the clinically used humerus T-plate (HTP), the cross-screw osteosynthesis (CSO), the unreamed proximal humerus nail with spiral blade (UHN), the recently developed Synclaw Proximal Humerus Nail (Synclaw PHN) and the angle-stable Locking Compression Plate Proximal Humerus (LCP-PH). The implant stiffness was determined for three clinically relevant load cases: axial compression, torsion and varus bending. In addition, a cyclic varus-bending test was performed to determine the implant properties under cyclic loading. RESULTS: In contrast to a rather elastic and minimally invasive implant(LCP-PH), the conventionally designed ones (Synclaw PHN, CSO, HTP, UHN) showed rather high stiffness values under static loading. In cyclic loading, a strong decrease in stiffness ( p<0.05) was found for the rigid implants HTP and UHN. In comparison with the other implants, only the elastic implant (LCP-PH) showed a significantly lower load reduction in a weak bone stock (17+/-6.2%). CONCLUSION: The high initial stiffness of rigid implants led to an early loosening and failure of the implant-bone interface under cyclic loading. Implants with low stiffness and elastic characteristics, however, appear to minimize the peak stresses at the bone-implant interface, making them particularly suitable for fracture fixation in osteoporotic bone. PMID- 12721685 TI - Do orthopaedic journals provide high-quality evidence for clinical practice? AB - BACKGROUND: In the hierarchy of research designs, randomized controlled trials and meta-analyses are considered to be evidence of the highest grade, and scientific journals are the main source of scientific information. METHODS: Using the National Library of Medicine Medline database, all randomized controlled trials and meta-analyses from 1966 to 1999 were retrieved from the journals indexed in the core list of the Science Citation Index in 1999, dedicated primarily to orthopaedics. The abstracts of the articles were reviewed independently by each author and classified by the year, journal name and subject. RESULTS: The total number of articles was 36,293, of which only 671 were randomized controlled trials (1.85%) and 12 were meta-analyses (0.03%). Although there was a progressively increasing trend for randomized controlled trials, more than half of them (81.9%) were published in four journals. Of the randomized controlled trials, 66% was about arthroplasty, and hip and knee arthroplasties covered 90.7%. CONCLUSION: Although the number of randomized controlled trials and meta-analyses is tending to increase, the conclusion of this study is that the high-quality evidence provided by the major orthopaedic journals is quite low, and more randomized controlled trials and meta-analyses are needed for evidence-based orthopaedic practice. PMID- 12721686 TI - Heat distribution and heat transport in bone during radiofrequency catheter ablation. AB - BACKGROUND: To assess the feasibility of percutaneous radiofrequency ablation in large bone tumours, the heat distribution in cortical bone and marrow around inserted electrodes was measured. METHODS: Fresh bovine cadaver tibial bones were locally heated through drill holes for a maximum of half an hour using water cooled single radiofrequency electrodes (Radionics Instruments Inc) by pulsed energy. Temperatures were measured in the marrow canal as well as in cortical bone by thermocouples at various distances from the inserted probes. RESULTS: Perpendicular to the probe, hyperthermia of more than 50 degrees C could be created in bone marrow in a sphere of approximately 3 cm, and of approximately 1 cm in cortical bone. CONCLUSION: As irreversible cellular damage can be expected when increasing the temperature to 50 degrees C for a duration of 6 min, this method may be effective for the minimal invasive ablation of neoplasms within human bone in cigar-shaped regions of approximately 3-cm diameter. PMID- 12721687 TI - Concomitant plantar tarsometatarsal (Lisfranc) and metatarsophalangeal joint dislocations. AB - We report an unusual case of concomitant plantar tarsometatarsal (Lisfranc) and 1st and 2nd metatarsophalangeal (MTP) joint dislocations and fracture of the neck of the third metatarsal bone which has never been reported before. The plantar dislocation of the Lisfranc joint was treated by open reduction and fixation with K-wires; the dislocations of the MTP joints and neck fracture of the third metatarsal bone were treated by closed reduction and percutaneous fixation with K wires and immobilized with a plaster cast. At the 5 year follow-up examination, our patient had no complaints, but the radiograph showed degenerative changes of the Lisfranc and the 1st MTP joint. PMID- 12721688 TI - Isolated peripheral neurilemoma attached to the tendon of the flexor digitorum longus muscle. AB - Neurilemomas are benign tumours arising from peripheral nerves with a Schwann cell sheath. They are normally painless and slowly growing, rarely causing motor disturbances. Neurilemomas are most common in the cranial nerves, in the trunk, the upper and lower extremities, but may appear anywhere. Especially rare are neurilemomas of the lateral peroneal nerve in the region of the fibular head and in the foot. We present the first detailed report of a neurilemoma localized between the Achilles tendon and the flexor digitorum longus muscle with separation of the tumour from the tibial nerve. PMID- 12721689 TI - Synovial involvement in hemangiomatosis. AB - BACKGROUND: Angiodysplastic lesions of the extremities are very often of great interest for the orthopedist, involving not only bones with length discrepancy, but also joints with hemarthrosis and synovial hypertrophy. METHODS: We describe 4 patients with cutaneous hemangiomatosis in the lower limb and a concomitant knee arthropathy. Like in other arthropathies, the articular damage is rapidly invasive and leads to progressive damage in the joint if not adequately diagnosed and treated. We treated the 4 patients at different ages, corresponding to different stages of intra-articular involvement. The preoperative management included MRI and diagnostic arthroscopy. All patients underwent an open synovectomy. All our patients were reviewed at a clinical and radiographic follow up after 10 years. RESULTS: In all patients, the range of motion was improved or at least not worsened after surgery, in spite of a larger extent of cutaneous lesions. X-ray and MRI confirmed clinical data showing no recurrence of the lesion inside the joint and no progression of the articular damage. CONCLUSION: The clinical outcomes were very good or good in the patients treated early, poorer in the patient operated on at an adult age. The importance of an early and precise diagnosis and of an accurate preoperative planning must be underlined. PMID- 12721690 TI - Complications of halo treatment for cervical spine injuries in patients with ankylosing spondylitis--report of three cases. AB - BACKGROUND: Patients suffering from ankylosing spondylitis are prone to injuries of the cervical spine even with minor trauma. Although the fractures are markedly unstable, nonsurgical treatment using a halo-thoracic plaster or jacket is a common approach. METHODS: We present three patients with cervicothoracic fractures of the ankylosed spine to describe problems and complications inherent in this type of treatment. In two, pin track infections and pin protrusion through the skull occurred, leading in one case to an intracerebral hemorrhage. In the third patient, the halo had to be removed after 8 months, just early enough to prevent the pins from cutting through. RESULTS: One patient required craniotomy. The second one could be resolved by local revision. In the third case, the fracture eventually united after using a stiff collar for 2 years. CONCLUSION: Halo treatment for cervical spine fracture in patients with ankylosing spondylitis is a challenging task for orthopedic surgeons and neurosurgeons. PMID- 12721691 TI - Periosteal ganglion in a child. AB - A case of periosteal ganglion in a 10-year-old boy is reported. The diagnosis of periosteal ganglion was made on roentgenographic and MR appearance and histologic evidence. The patient was successfully treated by excision. The follow-up examination showed no recurrence. Periosteal ganglia must be considered a cause of expanding cortical erosions. This lesion may mimic other peripheral expanding lesions. PMID- 12721693 TI - Pycnodysostosis associated with bilateral congenital pseudarthrosis of the clavicle. AB - We report the case of a 22-year-old woman with pycnodysostosis associated with bilateral congenital pseudarthrosis of the clavicle. The patient was first seen at the age of 19 years and had no symptoms during the 3-year follow-up period. The diagnosis of pycnodysostosis was made by typical clinical and radiological findings. Bilateral painless mid-clavicular mass and plain radiograms confirmed the diagnosis of bilateral congenital pseudarthrosis of the clavicle. Surgery was not performed for pseudarthrosis of the clavicles, just observation was preferred. PMID- 12721692 TI - Posterior dislocation of the shoulder associated with fracture of the humeral anatomical neck with 11-year follow-up after early open reduction and internal fixation. AB - BACKGROUND: . The combination of posterior dislocation and fracture of the anatomical neck represents an extremely rare injury. METHODS: . A patient with posterior dislocation of the shoulder and ipsilateral fracture of the humeral anatomical neck was treated with open reduction and internal fixation with two Kirschner wires. He was followed up for 11 years. RESULTS: . The functional results were excellent, and X-ray and MRI investigations revealed the absence of avascular necrosis of the humeral head. CONCLUSION: . Early and accurate open reduction with minimal osteosynthesis resulted in excellent function of the injured shoulder without avascular necrosis. PMID- 12721694 TI - Rapid sequence quadruple joint replacement in a rheumatoid Jehovah's Witness. AB - We report the case of a wheelchair-bound rheumatoid Jehovah's Witness who underwent rapid sequence, staged, simultaneous ipsilateral hip and knee replacements. Using perioperative erythropoietin and postoperative blood salvage, all four joints were reconstructed without homologous blood transfusion. One year following surgery, the patient has achieved an independent functional status, and all four replaced joints remain painless. PMID- 12721695 TI - Access to the medullary canal in closed antegrade femoral nailing: a technical report. AB - Although general recommendations exist regarding the correct placement of the skin incision and the direction of deep dissection for closed antegrade intramedullary nailing of the femur, in surgical practice simultaneously establishing the correct entry point and exact direction for insertion of the entry instrument in the lateral (sagittal) plane may be difficult. This is due to sub-optimal radiographic images in the lateral plane as a result of the overlying shadows of the pelvis, variations in the degree of rotation of the femur during patient positioning and fracture reduction manoeuvres, variations in the degree of anterior bowing of the femoral shaft and the anatomy of the greater trochanter, and deviations of the plane of deep dissection caused by the glutei muscle fibres. This may lead to the need for several attempts with increased damage to the glutei muscles, high exposure to radiation and the risk of an iatrogenic fracture. The present technical note describes a simple method for swift, easy and accurate access to the medullary canal during closed antegrade femoral nailing. PMID- 12721696 TI - Concerning Tero Heikkinen et al. (2002). Hemiarthroplasty or osteosynthesis in cervical hip fractures: matched-pair analysis in 892 patients. PMID- 12721700 TI - [Diffuse interstitial lung diseases in association with connective tissue diseases]. AB - This article gives a comprehensive overview of diffuse interstitial lung diseases especially those associated with connective tissue diseases. Interstitial lung diseases (ILD) represent a heterogeneous group of diseases of very different and partially unknown etiologies. Therefore, an exact classification of the ILD is difficult. ILDs are rarely seen in clinical practice and often lead to diagnostic problems. Diagnostic procedures require the patient's history, physical examination, serological-immunological and routine laboratory tests, pulmonary function testing, chest X-ray and high-resolution CT scan, bronchioloalveolar lavage (BAL) and bioptic procedures. The diagnostic approach to ILD requires an intensive multidisciplinary cooperation to offer early and effective therapeutic regimens. PMID- 12721699 TI - Features of cell death in brain and liver, the target tissues of progressive neuronal degeneration of childhood with liver disease (Alpers-Huttenlocher disease). AB - Alpers-Huttenlocher disease (AHD) is a rare encephalopathy of infancy and childhood characterized by myoclonic seizures and progressive neurological deterioration, usually associated with signs and symptoms of liver dysfunction. There is no biological marker of the disease, and ultimate diagnosis still relies on pathological examination. Features of clinical progression and pathological findings suggest AHD to be secondary to a genetically determined disorder of mitochondrial function. We report on four AHD patients and focus on their pathological features in brain, liver and muscle. Liver and muscle biopsy specimens were examined using histochemical markers of the oxidative pathways, probes to immunodetect molecules of the apoptotic cascades and electron microscopy. In liver (but not in muscle) biopsy samples, activated caspases were detected by immunohistochemistry: foci of caspase-9-positive cells were seen in a child affected with chronic, progressive fibrosis. In an 18-year-old boy, who suffered from valproic acid-associated acute hepatitis, caspase-3 cells were clustered among the necrotic foci and the foamy cells. In both patients electron microscopy revealed apoptotic nuclei. Normal muscle biopsy specimens were observed in two children, 2 and 8 years-old respectively; in the 18-year-old patient cytochrome oxidase-negative fibers as well as ultrastructural findings of mitochondrial abnormalities were observed. In no patient was there biochemical evidence of impaired oxidative metabolism. Neuropathological examination of the brains of two patients (13 months and 19 years old, respectively) showed focal distribution of the lesions affecting the telencephalic cortex and, to a lesser extent, subcortical gray nuclei. Along with the necrotizing lesions, characterized by neuronal loss, neuropil microcysts and newly formed vessels, we also observed acutely shrunken neurons and features of apoptotic cell death in the cerebral cortex only. Severe neuronal loss without necrotizing features was observed in the cerebellar cortex. The presence of both anoxic and apoptotic nuclei in brain and liver, the target tissues of the disease, is consistent with the hypothesis that abnormal activation of mitochondrion-related cell death pathways might be involved in the pathogenesis of AHD. PMID- 12721701 TI - [Imaging in collagen-vascular diseases with special emphasis on fibrotic lung alterations]. AB - This review presents the wide spectrum of radiological abnormalities seen in patients with collagen-vascular disorders. It provides an overview of both common and uncommon findings observed in this group of diseases. Moreover, diagnostic imaging algorithms used in conjunction with clinical tests are presented. In addition to conventional radiography and computed tomography (CT), these diagnostic algorithms also include more recent radiological modalities, such as thin section CT and multislice CT. PMID- 12721702 TI - [Heterogeneity of Borrelia burgdorferi: etiopathogenetic relevance and clinical implications]. AB - Of the ten different species of Borrelia (B.) burgdorferi sensu lato which have been characterized to date, only B. burgdorferi sensu stricto, B. garinii and B. afzelii have been identified as pathogenic in humans. It was suggested that different species possess different organotropisms and may preferentially cause distinct clinical manifestations of Lyme disease. Molecular analyses revealed a strong association of B. afzelii with the late cutaneous manifestation acrodermatitis chronica atrophicans, whereas B. garinii was predominantly identified in clinical samples from patients with neuroborreliosis. PCR-based analyses of samples from European patients with Lyme arthritis had given controversial results, but B. burgdorferi sensu stricto appears to be the major pathogen. The identity of the infecting species seems to be a major determinant in the pathogenesis of Lyme arthritis, although its complex immunopathological background and its clinical heterogeneity clearly indicate concomitant factors. Thus, characterization of the infecting organism at the species level on the one hand and linkage of clinical data with pathogenetically relevant immune parameters on the other, shall lead to a more precise understanding of the pathogenesis and the individual clinical course of Lyme borrelioses. PMID- 12721703 TI - Termination of disease-modifying antirheumatic drugs in rheumatoid arthritis and in psoriatic arthritis. A comparative study of 270 cases. AB - 102 rheumatoid arthritis (RA) and 104 psoriatic arthritis (PsA) patients' records were analysed according to a standardised protocol. Using Cox regression, life table analysis and log rank test, the effectiveness and toxicity of, and duration of disease modifying antirheumatic drug (DMARD) treatment were compared in RA and PsA. RA patients were treated with gold sodium thiomalate (GST), methotrexate (MTX) and sulphasalazine (SSZ) for a median duration of 35, 72 and 12 months respectively, whereas PsA patients were treated for 12, 12 and 17 months. The differences for GST and MTX were statistically significant (p=0.0043 and 0.0447). Drug toxicity was more frequently seen among patients with PsA (p=0.0023). No difference in efficacy could be proved. Results suggest that there is a significant difference between RA and PsA patients in terms of toxicity of these agents. Therefore, separate treatment strategies are needed, and earlier results with RA may not be directly applicable to PsA. PMID- 12721704 TI - [Multiple joint replacement of the lower limbs in rheumatoid arthritis]. AB - INTRODUCTION: Erosive-destructive changes in rheumatoid arthritis particularly in the large joints of the lower limbs often result in the patient's immobility. The aim of this retrospective study was to weigh the benefits of multiple joint replacement in rheumatoid arthritis against the specific complications. MATERIALS AND METHODS: From January 1977 until 31 December 1998, 215 patients with proven rheumatoid arthritis (according to the ACR classification) received three or four endoprostheses in the lover limbs. In a retrospective study with a standard questionnaire and evaluation of the patient record, we were able to assess 201 rheumatoid patients (93.5%) who received 688 endoprostheses (93.7%), with 347 total hip prostheses (THR) and 341 total knee prostheses (TKR). RESULTS: There was a significant improvement of the patient's ability to walk and in 16.9% of the cases even a remobilization of the rheumatoid patients who had been unable to walk before surgery. On the other hand, there were significant complications--in the majority of the cases (appr. 60%) with total knee replacement. Together the complications per implant had an extremely high complication rate of 43.8% (n=88) per rheumatoid patient. DISCUSSION: Due to the long-term follow-up of 22 years (1977 through 1998), we were able to demonstrate that with a new implant generation and a more a careful indication this unacceptable complication rate can be reduced considerably. However, the multiple joint replacement of the lower limbs remains a difficult method with a high complication rate and should therefore only be performed at orthopedic rheumatological centers. PMID- 12721705 TI - Validation of a German version of the 'Disabilities of Arm, Shoulder and Hand' questionnaire (DASH-G). AB - OBJECTIVE: In 1994 the American Academy of Orthopedic Surgeons' outcome research committee developed and validated a functional outcome questionnaire for disabilities of the upper extremity (DASH). The objective of our study was to translate the DASH into German and to evaluate its reliability and validity for German-speaking patients with shoulder pain. METHODS: Translation-backtranslation of the DASH was performed according to published guidelines. Psychometric properties and validity were assessed in 49 consecutive patients with shoulder pain originating from within the shoulder girdle. Age, duration of symptoms and current pain were obtained. Test-retest reliability was assessed in a subset of 18 patients who filled in the questionnaire 10 days later. Internal consistency was evaluated with Cronbach's alpha coefficient. Test-retest reliability was assessed using Spearman correlations and the Bland and Altman plot. To study the validity, we examined the correlation of the DASH with other measures of function of the upper extremity including the HAQ subscales relating to the upper extremity and pain measures (e.g. numerical rating scale, SF-36 pain scale). We also examined our hypothesis that the DASH is moderately correlated with measures of range-of-motion (e.g., forward flexion and abduction). RESULT: Translation backtranslation revealed no major difficulties. The mean age of the patients was 59 years, mean duration of symptoms 60 months and current pain score 5.6 (NRS). Test-retest reliability was 0.90 for the total DASH. The internal consistency was 0.96. Strong correlations (p<0.01) were found between the DASH and the mean among the five subscales of the HAQ representing upper extremity function (0.88) and pain as measured with the SF-36 bodily pain scale (-0.79). As hypothesized, the DASH was also moderately correlated with measures of range-of-motion (e.g. forward flexion rho=-0.49; abduction rho=-0.57). CONCLUSION: Our data confirm that the German version of the DASH retains the characteristics of the American original and is a reliable and valid instrument to measure functional disability in German speaking patients with shoulder pain. PMID- 12721706 TI - [Muscle weakness and Gottron's sign]. PMID- 12721707 TI - [Cervical myelopathy induced by calcium pyrophosphate tophus in primary chondrocalcinosis]. AB - A 78 year old female patient with cervical myelopathy induced by a calcium pyrophosphate dihydrate tophus around the dens axis with clinical signs of a cervical tumor is presented. After operation of the tumor, the diagnosis of a generalized CPPD disease was established retrospectively, where by patient had complained of joint pain already for many years. This known complication of a primary chondrocalcinosis should be a reason for a careful investigation of the cervical spine in CPPD disease, because neurological disturbances might increase the joint destruction in the manner of "Charcot joints". PMID- 12721708 TI - [Pigmented villonodular synovitis of the hip--diagnosis and therapy of an invasive disease]. AB - In the diffuse form of pigmented villonodular synovitis, the transformed synovia invades local bone and cartilage, leading to destruction of the osseous and cartilage tissue. Secondary osteoarthritis and ultimately joint destruction is a result. Malignant development is under discussion. The knee is the most frequently involved joint, followed by involvement of the hip, which is well documented in several studies. This paper introduces a case of pigmented villonodular synovitis involving the hip. Its general diagnostic features and treatment options are introduced and discussed, including the role of reconstructive joint surgery in advanced cases of PVS. The key function of early diagnosis and treatment resulting in joint-salvaging procedures is emphasized. PMID- 12721709 TI - The role of laparoscopy in the management of lower gastro-intestinal bleeding. AB - This report describes three cases of significant lower gastro-intestinal haemorrhage caused by a bleeding Meckel's diverticulum. In the first two cases a pre-operative technetium-pertechnetate or Meckel's scan was negative or inconclusive, and in the third case no Meckel's scan was carried out. The diagnosis was established at laparoscopy in all three cases and in each case the diverticulum was excised extracorporeally. PMID- 12721710 TI - Effect of a high fat diet on lipid absorption and fatty acid transport in a rat model of short bowel syndrome. AB - Long chain fatty acids (LCFAs) appear to be powerful stimulants for small bowel adaptation in patients with short bowel syndrome (SBS). However, the dietary lipid content may alter intestinal lipid transport. The aim of this study was to investigate the effects of a high fat diet (HFD) on in vivo lipid absorption and molecular and cellular mechanisms of LCFAs uptake by the remaining bowel. Male Sprague-Dawley rats (240-280) were randomly assigned to one of three groups: sham rats fed normal chow (sham-NC), SBS rats fed NC (SBS-NC) and SBS rats fed HFD (SBS-HFD). SBS rats underwent a 75% small bowel resection. Rats were sacrificed on day 3 or 14. Body weight, fat intake and fat clearance (total fecal fat) were measured twice a week. Fat absorbability was calculated as intake minus clearance and was expressed as percent of intake. Total RNA from the mucosa of duodenum, jejunum and ileum was extracted using TRIZOL Reagent. Northern blot analysis was performed to determine FAT/CD36 mRNA levels. Enterocyte LCFA transport was measured on day 14. LCFA uptake was determined by measuring cellular [3H]-oleate uptake over time (4-120 s). Mean (+/-SE) FAT/CD36 mRNA levels and oleate uptake kinetic parameters were analyzed using ANOVA. Fat absorbability diminished after bowel resection, suggesting fat malabsorption. Remaining bowel in SBS-NC rats responded by an increase in FAT/CD36 mRNA levels in the duodenum and ileum on day 3, and the duodenum and jejunum on day 14 compared to sham-NC animals, and was accompanied by an increase in enterocyte LCFA transport in all segments. Exposure to a HFD for 14 days resulted in significantly increased fat absorbability after 3 days compared to SBS-NC rats. However, FAT/CD36 mRNA levels (vs. SBS-NC) decreased in all segments on day 3. On day 14, FAT/CD36 mRNA levels were decreased in the duodenum and ileum and were accompanied by reduced oleate uptake by isolated enterocytes in the ileum (vs. SBS-NC). In a rat model of SBS, early high fat diet increased lipid absorptive capacity of the intestinal remnant as seen by increased fat absorbability. The main mechanisms of this effect may be an acceleration of structural intestinal adaptation resulting in an increased number of enterocytes. However, at molecular and cellular levels HFD decreased mucosal FAT/CD36 mRNA levels and oleic acid uptake by isolated enterocytes. PMID- 12721711 TI - Appendicitis in infancy. AB - Acute appendicitis is the most common cause of abdominal pain requiring surgery in children. But it is an uncommon entity in young children and rare in infants. During a 10-year period, between January 1991 and December 2000, 7 infants (age from 17 days to 8 months) were treated for acute appendicitis at Changhua Christian Hospital. All of the preoperative symptoms and signs, the duration between admission and operation, pathology reports, and laboratory data were reviewed. There were no specific clinical signs and symptoms. The duration between admission and operation ranged from 2 hours to 5 days (mean: 31.4 hours). The pathology reports revealed 3 were gangrenous, 3 were gangrenous with perforation and one was suppurative. There was one death. Pseudomonas aeruginosa was cultured from blood, ascites, bile or stool in 6 cases (85.7%). Early diagnosis of acute appendicitis in infants is still difficult. Although the mortality has declined, the morbidity still remains high. The high percentage of infection by Pseudomonas aeruginosa should be further evaluated in infantile appendicitis. PMID- 12721712 TI - Investigations for incontinence and constipation after surgery for Hirschsprung's disease in children. AB - Surgery for Hirschsprung's disease is associated with high rate of morbidity, in the form of either constipation or incontinence or a combination of the two. This study investigates the mechanisms responsible for incontinence and/or constipation following the pull-through operation for Hirschsprung's disease. There were 19 children (15 boys and 4 girls), who at the time of study; 16 had undergone Duhamel, 1 Rehbein, and 2 Soave operation. We classified patients according to their symptoms into 3 groups: Group A was incontinent of faeces; Group B was constipated and incontinent of faeces, and Group C was constipated only. The median age at referral was 6 years, and the median period after operation was 5 years. All patients were investigated by intestinal transit study, endoanal sonography and anorectal manometry. Group A had normal or rapid transit study, as opposed to Groups B and C, who had delayed-transit study. On endoanal sonography, all children had an intact internal and an external anal sphincter, below the level of pull-through operation. The anorectal manometry showed a significantly lower resting anal pressure in the incontinent Group A as compared to the constipated children with or without incontinence in Group B or C (38 mmHg versus 57 or 66 mmHg respectively). The rectal pressure was also significantly higher in children in Group A as compared to those in Group B or C (71 mmHg versus 42 or 36 mmHg). The ratio of rectal/anal pressure was higher in incontinent children in Group A, as compared to constipated children in Group B or C. Therefore, constipation can be caused by high anal resting pressure and a weak rectal peristalsis, while faecal incontinence can be secondary to poor compliance and elevated rectal pressure in the presence of normal or low anal sphincter resting pressure. Aperients are the mainstay of treatment of constipation, however, children with incontinence are more difficult to treat. We did not attempt to define the pattern of nerve plexus because of poor results of revision operation for residual hypoganglionic segment and intestinal neuronal dysplasia. Treatment of these children can become more rational, if furnished with detailed functional studies. We advocate investigation of the anorectal function at an early stage in symptomatic children after surgery for Hirschsprung's disease, and less invasive treatment should be considered before embarking on major surgery. PMID- 12721713 TI - Application of the Meckel's scan in a case of gastric corrosive injury. AB - This report describes a useful application of (99m)Tc-pertechnetate abdominal scintigraphy (Meckel's scan) in the assessment of gastric viability after corrosive injury. The Meckel's scan provided information about residual gastric mucosal function and prognosis in a less invasive manner than more usual methods. PMID- 12721715 TI - Congenital colonic atresia: should primary anastomosis always be the goal? AB - We report 4 cases of the rare condition, congenital colonic atresia, presenting over 9 years. Two patients had dilated loops of bowel noted on routine ante-natal ultrasound. Three had primary anastomosis for lesions in transverse or distal descending colon and one had a staged procedure with colostomy formation. All had type III atresia. One patient had an early leak following primary colo-colic anastomosis for atresia extending from the hepatic flexure to the sigmoid colon. If it is desired to preserve the proximal colon in a right sided lesion with significant loss of colonic length then primary anastomosis may not be safe and we would advocate a staged procedure. Otherwise we would support current moves towards primary anastomosis in this condition. We also urge early investigation in all patients who exhibit intestinal dilatation on antenatal ultrasound. PMID- 12721714 TI - Carbon dioxide laser circumcisions for children. AB - The carbon dioxide laser for circumcision was introduced by our department in 1989. This study aims to review our experience with laser circumcision for children and to evaluate its cost effectiveness as compared to conventional methods. A retrospective study of 30 patients who underwent conventional circumcision in 1985 and another 30 patients who underwent laser circumcision in 1995 was undertaken. The operating times in both groups were compared. The total cost of use of the laser machine was calculated, taking into account maintenance costs, estimated life span of laser machines (10 years) and costs of disposables used during each circumcision. This was weighed against the cost savings from shorter operating times and reduced operating theatre facility charges. Also, morbidity data from 2781 laser circumcisions done between May 1997 and April 2000 was collected. There was a significant decrease of 5 minutes in operating time for the group of patients who underwent laser circumcision. Calculated cost savings per laser circumcision from the reduced operating theatre time was S dollars 31/-. Of the 2781 cases of laser circumcision performed, there was an overall complication rate of 1.15%. Twenty-nine cases (1.04%) had post circumcision bleeding, of which 10 cases (0.36%) required unplanned return to operating theatre for hemostasis. Three cases (0.11%) had wound infection, requiring admission to hospital. Laser circumcision is a simple method with reduced operative time translating into cost effectiveness. Morbidity rates of laser circumcision compare favourably to those of conventional circumcision based on reports from other institutions. PMID- 12721716 TI - A prospective comparison of tissue glue versus sutures for circumcision. AB - The safety of wound closure with tissue glue after surgery has been well established. The efficacy of its use in circumcision is poorly documented. The aim of this study was to carry out a comparative study of tissue glue versus suture closure after circumcision. The duration of the operative procedure, pain score, cosmesis and postoperative complications were evaluated. There were no complications in either group. There was no statistically significant difference in the pain score in both groups. However the mean time taken for tissue glue was 16.6 minutes and the mean time taken for sutures was 23.7 minutes. (p < 0.0001) which was statistically significant. The cosmetic appearance was found to be superior in the tissue glue group as there were no suture marks on the join of the shaft skin and foreskin base. PMID- 12721717 TI - The detection of IgG antibodies to silicone. AB - Following a recent report of an ELISA test for the detection of antibodies to silicone, we attempted to use the same assay in four patients with known exposure to silicone. These patients all gave similar positive results as did a number of control sera with no known silicone exposure. We conclude that this assay does not measure serum levels of antibodies to silicone. PMID- 12721719 TI - Management of blunt hepatic injury in children: usefulness of emergency transcatheter arterial embolization. AB - We reviewed our experience to determine the usefulness of emergency transcatheter arterial embolization (TAE) for severe blunt hepatic injury (BHI) in children. Between 1978 and 2000, 21 children with BHI (14 boys and 7 girls, ranging in age from 2 to 14 years) were managed according to our protocol. The patients who were hemodynamically stable, and had no other associated injury requiring laparotomy, regardless of the hepatic injury grade, were managed nonsurgically. Emergency angiography and TAE performed after a CT scan revealed extravasation of the contrast medium. Of the 21 patients, 3 underwent emergency laparotomy; 2 due to hemodynamic instability despite fluid resuscitation (1 died), and the 3rd patient because of associated injury. The other 18 patients (86%) were initially managed nonsurgically; however, 2 underwent delayed laparotomy because of complications (1 each of suspected delayed hepatic hemorrhage and liver abscess). Nonsurgical management was completed in the remaining 16 (89%) with no morbidity and mortality. Two of the 16 returned to a hemodynamically stable condition with fluid resuscitation, but were compromised with persistent hepatic hemorrhage, and were successfully treated with emergency TAE. We propose that emergency TAE should be considered as an initial treatment for severe BHI in children. PMID- 12721720 TI - Renal and peri-renal abscesses in children: proposed physio-pathologic mechanisms and treatment algorithm. AB - Renal and perinephric abscess in children are uncommon. Three basic pathophysiologic mechanisms are involved, namely, hematogenous spread, ascending infection and contamination by proximity to an infected area. Six pediatric patients diagnosed with renal abscess were treated at our institution from 1990 2000. Five patients were females; ages ranged from 3-17 years (mean 11.8 years). Diagnosis, as expected, was not readily apparent at presentation. Computerized tomography and renal sonograms were the most useful imaging modalities. Gram negative bacteria were commonly isolated; only one patient grew Staphylococcus aureus. All patients received broad-spectrum intravenous antibiotics. Additional treatments consisted of percutaneous drainage (4 patients), exploratory laparotomy (1 patient, for presumed Wilm's tumor) and nephrectomy (2 patients). A new classification of the etiologic mechanisms of this condition is proposed along with a simple and practical treatment algorithm. PMID- 12721718 TI - Matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1: expression in the lung of fetal rats with nitrofen-induced diaphragmatic hernia. AB - The surrounding extracellular matrix of airway wall tissues changes in response to mechanical stresses and hypoxia. The presence of matrix metalloproteinase-9 (MMP-9) and its inhibitor, tissue inhibitor of metalloproteinase-1 (TIMP-1), is correlated with collagen degradation and tissue repair in lung disorders. The aim of this study was to evaluate the expression of MMP-9 and TIMP-1 in the lung of fetal rats with nitrofen-induced congenital diaphragmatic hernia (CDH). Administering 100 mg of nitrofen dissolved in 1 ml olive oil to pregnant Wistar rats on day 9 of gestation induced left-sided CDH in fetal rats. In control animals, the same dose of olive oil was given without nitrofen. Cesarean section was performed on day 21 of gestation. The fetuses were divided into two groups: normal controls (n = 10) and nitrofen-induced left-sided CDH (n = 10). Immunoreactivity of the staining for MMP-9 and TIMP-1 in the lung tissues was semiquantitatively analyzed using the staining scores. The relative amount of MMP 9 or TIMP-1 divided by the amount of beta-actin for each lung sample was measured by using the real-time reverse-transcriptase polymerase chain reaction. The immunoreactivity of MMP-9 was significantly increased in the CDH group (n = 5) compared with the control group (n = 5) (p = 0.031). On the other hand, the immunoreactivity of TIMP-1 in the two groups was not significantly different (n = 0.134). The relative amount of MMP-9 (or TIMP-1) in the CDH group (n = 5) does not differ significantly from that in the control group (n = 5) (p = 0.059, 0.596, respectively), but the relative amount of MMP-9 is higher in the CDH group, although it is not significantly higher. On the other hand, the ratios of MMP-9 to TIMP-1 were significantly higher in the CDH group (p = 0.028). In conclusion, fetal rats with nitrofen-induced CDH, a model of respiratory disorders, manifested the excess of MMP-9 activity due to the absence of TIMP-1 that would suggest a trend toward disruption of the extracellular matrix in the CDH lung tissues. PMID- 12721721 TI - Long-term evaluation of esophageal function in patients treated at birth for esophageal atresia. AB - Dysphagia, gastroesophageal reflux (GER) and esophageal metaplasia are reported with various incidence in the long term follow-up of patients treated at birth for esophageal atresia (EA). To evaluate the long term outcomes 26 patients treated at birth for EA with Tracheo Esophageal Fistula (TEF) were examined 8-28 (mean 15.8) years later by clinical evaluation, including barium meal, fiberoptic upper GI endoscopy, 24 hour ambulatory two-channel pH-monitoring and stationary esophageal manometry. 50% of patients complained of dysphagia. Mild esophagitis was found in 20% of patients but GER was detected in only 16.7% of the cases. By morphological X-ray, esophageal anomalies were detected in 31% of cases without significant functional relevance. Hundred percent of patients had a disorganized peristaltic esophageal activity and a low amplitude of the esophageal contractions was observed in 58% of them. In our series, esophageal dismotilty seems to be the main consequence of EA without any relevant disturbance of normal nutritional habit. PMID- 12721722 TI - Treatment of intestinal pseudo obstruction by segmental resection. AB - Intestinal pseudo-obstruction refractory to medical therapy is a debilitating problem for specialists dealing with gastrointestinal disorders. We report the case of a newborn who developed severe, recurrent symptoms of intestinal obstruction, due to visceral myopathy. The case was persistently intractable to medical management, leading to repeated laparotomies. Gastrointestinal lesions showed marked dilatation of the entire digestive tract, with enlarging to enormously distended segments at two areas. Resection of these segments improved bowel function, facilitating enteral nutrition. Long-term hyperalimentation and repetitive hospitalizations were also avoided with this procedure. These results suggest that segmental resections can save unnecessary intestinal resections in cases with extensive gastrointestinal involvement. PMID- 12721723 TI - Sacrococcygeal teratomas: the UK Children's Cancer Study Group's experience. I. Neonatal. AB - The aim of this study was to review the United Kingdom Children's Cancer Study Group (UKCCSG) experience of sacrococcygeal teratomas (SCT) including histological presentation, response to surgery and chemotherapy, and long term effects of the tumour and treatment. This paper presents the results for those children diagnosed during the neonatal period. Children aged up to 4 weeks with biopsy proven localised or metastatic sacrococcygeal germ cell tumours were eligible. From 1st January 1989 to 31st December 1997 (9 years), 15 UKCCSG centres registered 51 neonates with SCT into GC 8901. Surgery alone was performed in all and the prognosis was good - except for 1 baby who died from massive haemorrhage at the initial operation and 1 who died from the complications of prematurity. Seven of the 51 children (14%) who had teratomas in the neonatal period (5 mature, two immature) had yolk sac tumour (YST) recurrence at: 4, 12, 15, 20, 20, 28 and 32 months of age. These children received chemotherapy in the form of etoposide/bleomycin/carboplatin (JEB) and are alive and well at review. These results emphasise the need for oncological follow-up of SCT and the good response to JEB chemotherapy of malignant teratomas and YST. PMID- 12721724 TI - Proposal of fibrosis index using image analyzer as a quantitative histological evaluation of liver fibrosis in biliary atresia. AB - This study was designed to elucidate whether the fibrosis index (FI), which was measured as a ratio of histological fibrotic tissue area to the whole area using an image analyzer, could reflect liver function and long-term prognosis in biliary atresia (BA). Liver biopsies were performed in 46 BA patients at hepatic portoenterostomy (HPE) and stoma closure. The chronological difference rate of FI (FIDR) was the monthly FI difference between HPE and stoma closure. FI at HPE and stoma closure was significantly higher than in the control. FI at HPE and at stoma closure significantly correlated with gammaGTP or T.Bil, D.Bil, cholinesterase and total bile acid, respectively. FIDR in jaundice-free group was significantly lower than in jaundiced group at 5 years after HPE. FIDR in V-2 (varices with red-color sign) was significantly higher than in V-0 (no varices) or V-1 (varices without red-color sign). ICG-K value significantly correlated with FIDR. FI at stoma closure or FIDR was significantly lower in living patients than in patients who eventually died or underwent liver transplantation. In conclusion, FI can reflect the degree of cholestasis in BA. FIDR would be useful for predicting long-term outcome in BA. PMID- 12721725 TI - When to resect and when not to resect an asymptomatic Meckel's diverticulum: an ongoing challenge. AB - To determine the morbidity and mortality of Meckel's diverticulum (MD) as a cause of acute abdominal disorders and to evaluate the relationship between patient age, MD complications, and postoperative complications. We reviewed 74 patients who underwent surgery between 1990 and 2000 for an acute abdominal syndrome with a MD diagnosed intraoperatively. Forty children were treated before 1995 and reviewed retrospectively, while the remaining 34 were reviewed prospectively. The average age was 4.8 years; the male/female ratio was 2.5/1; 34 (46%) were less than 2 years old, 32 were between 2 and 8 years, and 8 were older than 8 years. None of the symptoms was suggestive of the diagnosis of MD. Thirty-nine MDs were asymptomatic (21 intussusception, 18 volvulus), but all were the secondary cause of the acute abdomen. The remaining 35 children had a symptomatic MD (diverticulitis in 14, diverticular bleeding in 11, diverticular perforation in 10). The risk of complications due to a MD occurring in children under 2 years and between 2 and 8 years of age was significantly higher compared to children older than 8 years (P = 0.02). Postoperative complications occurred more commonly in children between 2 and 8 years of age compared to other patients. There is thus an increased risk of morbidity in a symptomatic MD in patients less than 2 and between 2 and 8 years of age, and there is no predictive factor for the development of diverticular complications. Resection of the MD is recommended in all children younger than 8 years, including asymptomatic ones, in the absence of absolute contraindications. PMID- 12721726 TI - Multiple endocrine neoplasias type 2A and thyroid medullary carcinoma: an interdisciplinary challenge. AB - The diagnosis and treatment of multiple endocrine neoplasias type 2A (MEN 2A) requires interdisciplinary management. The association of RET proto-oncogene mutations and medullary thyroid carcinoma (MTC) in children is well-known, but the optimal timing for elective surgery is controversial. Besides the risk of MTC, associated anomalies like hyperparathyroidism have to be considered. We report the results of molecular genetic investigations, the pentagastrin stimulation test, pre- and postoperative staging, and histologic examinations of four children who had a positive family history for MEN 2A. Histologic specimens of the removed thyroid glands showed MTC in all four cases. The patients had an uneventful postoperative clinical course. In view of the recent literature and our patients' results, we suggest a concept for diagnostic strategy and timing of the elective thyroidectomy. PMID- 12721727 TI - Acalculous cholecystitis in Nigerian children. AB - Sixteen children with acalculous cholecystitis (AC) were treated over a 9-year period (13 male and 3 female). Their ages ranged from 8 to 18 years (median 11). Eight (50%) presented with complications (perforation 4, gangrene 2, empyema 2); 13 (80%) presented with acute AC with a duration of symptoms of 2 weeks or less while 3 (20%) presented with chronic AC with symptoms present for more than 3 months. The diagnosis was made by ultrasound except in the patients with complications, who were diagnosed at laparotomy. Salmonella typhi was cultured in the bile and blood in 2 cases and the Widal titre was significantly elevated in 4 others. One child had chronic blockage of the cystic duct by a lymph node; in 9 there was no identifiable cause. Open cholecystectomy was successfully performed in 15 cases, while 1 child was managed non-operatively. The need for early diagnosis of cholecystitis in children is obvious if the potentially life threatening complications of perforation and gangrene are to be avoided. PMID- 12721728 TI - A prospective study of neonatal inguinal herniotomy: the problem of the postoperative hydrocele. AB - Previous studies have reported an increased incidence of complications following neonatal inguinal herniotomy (IH) in boys. The incidence and natural history of postoperative hydrocele in such cases has not been described. A prospective follow-up study of a consecutive series of male infants weighing less than 3 kg at the time of IH was undertaken. Regular follow-up examinations were scheduled for at least 1 year. Thirty-eight boys weighing less than 3 kg underwent IH during an 18-month period. One subsequently died from complications of prematurity. Complete follow-up data were available for 29/37 (78%) patients, yielding a total of 46 IHs. There were 2 recurrent hernias (4%), 2 unequivocally atrophic testes (4%), and 1 iatrogenic testicular ascent (2%). Five ipsilateral hydroceles complicated the postoperative course of 4 boys (14% of patients, 11% of herniotomies). Two of these were explored, but in neither case was a recurrent/residual patent processus vaginalis found. One hydrocele was aspirated without recurrence, and the remaining 2 resolved spontaneously. The complication rate in small infants undergoing neonatal IH is significantly higher than in older boys. The presence of a hydrocele after neonatal IH may simply reflect the accumulation of fluid in the distal hernia sac and, provided there is no evidence of a recurrent inguinal hernia, an expectant approach is recommended. PMID- 12721729 TI - Should cholecystectomy be performed concomitantly with splenectomy in children with sickle-cell disease? AB - Splenectomy and cholecystectomy are among the common surgical procedures required to treat complications of sickle-cell disease (SCD), and when performed separately have been shown to be safe and effective. To determine whether cholecystectomy be performed concomitantly with splenectomy (CSC) in these children, we studied a total of 130 children who underwent splenectomy for various hematologic diseases at our hospital. The most common indication was SCD. Ninety-nine patients (86 SCD and 13 sickle-B-thalassemia) underwent splenectomy and 19 (19.2%) (12 males and 7 females, mean age 13.4 years [7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18]) underwent CSC due to concomitant gallstones, which were asymptomatic in 13 cases. Those with an admission hemoglobin (Hb) of less than 10 g/dl were transfused with packed erythrocytes to increase their Hb to 10-12 g/dl and their hematocrit to 30%-40%. All patients received intravenous hydration the night before surgery at a rate of 1(1/2) times their maintenance rate, which was continued postoperatively until they were able to tolerate adequate oral fluid intake. The indications for splenectomy in those who had CSC were: acute splenic sequestration crisis in 12, splenic abscess in 3, hypersplenism in 2, and massive splenic infarction in 2. Eight patients had massive splenomegaly (spleen weight >1 kg). In 7 cases CSC was done through a left-upper-quadrant (LUQ) transverse incision, in the remaining 12 through an upper midline incision. There was no mortality and only 2 patients developed postoperative complications; a wound infection in 1 and a hematoma in the splenic bed in another. With good perioperative management, CSC is both safe and effective for children with SCD. Both procedures can be performed safely through an upper midline or a LUQ transverse incision, even in the presence of massive splenomegaly. PMID- 12721730 TI - The impact of cystic fibrosis on neonatal intestinal obstruction: the need for prenatal/neonatal screening. AB - To determine the incidence of cystic fibrosis (CF) in neonates with intestinal obstruction (NIO) secondary to meconium ileus (MI), jejunoileal atresia (JA), meconium plug syndrome (MPS), volvulus (V), and meconium peritonitis (MP) and analyze the correlation of ultrasonographic (US) signs with CF in NIO with a prenatal diagnosis of intestinal anomaly, a prospective analysis of different types of NIO from 1990 to 1998 was undertaken. Immunoreactive trypsin measurement, genetic studies, and sweat tests were performed to confirm or rule out CF. Cases with prenatal diagnosis were analyzed for gestational age, dilated bowel, ascites, hyperechoic bowel, and calcifications. Of 80 neonates, 19 (24%) had CF: 2/33 (6%) JA, 6/14 (43%) MPS, 1/14 (7.1%) MP, 10/10 (100%) MI, and 0/9 V. Thirty (37.5%) had a prenatal diagnosis of an intestinal anomaly. The overall incidence of CF in NIO with a prenatal diagnosis of intestinal anomaly was 4/30 (13%), or 333 times the estimated risk of CF in the general population. A hyperechoic pattern with dilated bowel was associated with higher specificity for CF: 3/3 cases (100%), followed by hyperechoic bowel with ascites: 3/4 cases (75%). All babies with any type of NIO should thus be screened for CF. Prenatal screening for CF should be indicated in all pregnancies with US patterns of specific intestinal disorders. PMID- 12721731 TI - Bilateral phrenic-nerve paralysis treated by thoracoscopic diaphragmatic plication in a neonate. AB - Traditionally, diaphragmatic plication (DP) is performed via a thoracotomy that includes incision of the lower intercostal muscles, which are involved in respiratory movement. This may adversely affect ventilation by causing deterioration of respiratory function and making ventilation less efficient. These problems do not occur with thoracoscopic DP (TDP), since the lower intercostal muscles are left intact. We describe a full-term newborn infant with bilateral phrenic-nerve paralysis (PNP) who was treated by TDP and conclude that this method may be effective in the treatment of PNP even in small infants. PMID- 12721732 TI - Use of parenteral testosterone prior to hypospadias surgery. AB - Surgical correction of genital defects was formerly proposed when the size of the penis was sufficient to permit easy surgical repair. To enlarge penile size, temporary stimulation with testosterone or dihydrotestosterone cream has been used; however, the results were not only inconsistent, but absorption was also variable. We report our experience with parenteral testosterone as an adjunct to reconstructive genital surgery in 25 patients aged 6-18 months from July 1999 to December 2000, including 8 with penile hypospadias, 15 with penoscrotal hypospadias, and 2 with perineal hypospadias. Each had a penis that was significantly smaller than usual. Testosterone enanthate 25 mg was given i.m. once per month for a total of three doses before surgical repair. Penile length and glans circumference were measured before therapy and at operation. Side effects such as the development of pubic hair and acne were monitored. Bone age was checked 1 year later. An increase in penile length (from 19.8 +/- 2.4 mm to 23.8 +/- 2.0 mm) and glans circumference (from 27.4 +/- 1.4 mm to 37.84 +/- 2.6 mm) was apparent in all except 2 patients (P < 0.001 for both, paired t-test). Four patients had a significant increase in either penile length or glans circumference after the initial dose so that no further injections were required. No definite secondary effects were found. Preoperative parenteral testosterone therapy thus causes a significant increase in penile length and glans circumference without apparent side effects. We suggest that this therapy prior to microphallic hypospadias repair is appropriate. PMID- 12721733 TI - Unusual intestinal sequelae after operations for Ascaris lumbricoides infestation. AB - Two children with severe bowel sequelae distal to the site of an Ascaris lumbricoides bowel obstruction are reported. It is postulated that these resulted from inflammatory reaction to toxic decomposition products of disintegrating worms. Surgery should be modified in the presence of bowel-wall injury distal to the site of ascarid worm - bolus obstruction. PMID- 12721734 TI - The effect of pentoxifylline on intestinal anastomotic healing after ischemia. AB - The negative effects of ischemia-reperfusion injury (IRI) on intestinal anastomotic healing have been described in previous studies. To determine the effect of pentoxifylline (PTF) on small-bowel anastomoses performed after IRI, 20 male Wistar albino rats were studied. Under ketamine anesthesia, the superior mesenteric artery was occluded for 30 min and ileal resection-anastomosis was performed after 10 min of reperfusion. In group I, no medication was given. In group II, PTF 50 mg/kg was administered intraperitoneally before reperfusion and i.m. on the 1st, 2nd, 3rd and 4th postoperative days (POD). All rats were killed and anastomotic segments were removed on the 7th POD. Anastomotic bursting pressure (ABP) and hydroxyproline (HP) content were measured in the anastomotic segments; parameters were compared statistically with Student's t-test. In group I, the mean ABP was 225.5 +/- 38.18 mmHg the and mean HP level was 16.03 +/- 1.91 micro mol/g tissue. In group II, these levels were 277 +/- 41.1 mmHg and 19.85 +/ 1.81 micro mol/g tissue, respectively. The differences between the groups were significant (P < 0.005). Thus, PTF can improve anastomotic healing in an experimental model after IRI by promotion of collagen synthesis. PMID- 12721735 TI - Mutilations due to medical disorders in children. AB - Soft-tissue and bone necrosis, although rare in childhood, occasionally occur in the course of infectious diseases, either viral or bacterial, and seem to be the result of hypoperfusion on a background of disseminated intravascular coagulation. Treatment consists in correction of septic shock and control of necrosis. Necrosis, once started, shows extraordinarily rapid evolution, leading to soft-tissue and bone destruction and resulting in anatomic, functional, psychological, and social handicaps. Ten mutilated children were treated from January 1986 to January 1999 in Hospital de Dona Estefania, Lisbon, Portugal. One was recovering from hemolytic-uremic syndrome with a severe combined immunodeficiency, another malnourished, anemic child had malaria, and three had chicken pox (in one case complicated by meningococcal septicemia). There were three cases of meningococcal and two of pyocyanic septicemia (one in a burned child and one in a patient with infectious mononucleosis). The lower limbs (knee, leg, foot) were involved in five cases, the face (ear, nose, lip) in four, the perineum in three, the pelvis (inguinal region, iliac crest) in two, the axilla in one, and the upper limb (radius, hand) in two. Primary prevention is based on early recognition of risk factors and timely correction. Secondary prevention consists of immediate etiologic and thrombolytic treatment to restrict the area of necrosis. Tertiary prevention relies on adequate rehabilitation with physiotherapy and secondary operations to obtain the best possible functional and esthetic result. PMID- 12721736 TI - Evaluation of the protective effect of methotrexate on OK432-induced liver injury in a rat model. AB - Cholangitis-induced liver damage was created in rats by intraductal injection of OK432, a lyophilized incubation mixture of group A Streptococcus pyogenes of human origin. Oral, low-dose methotrexate (MTX) was given to one group to study its protective effect on liver injury. The liver histology was studied and semiquantitatively graded. The OK432-induced changes were compared with and without MTX therapy. The results revealed statistically significant higher grades of portal inflammation and sinusoidal infiltration in rats treated with OK432 compared to saline-treated controls. There was significant improvement in liver changes in the group treated with MTX compared to the untreated group. It could be concluded that oral, low-dose, pulsed MTX therapy caused significant improvement in cholangitis-induced liver damage in rats. PMID- 12721737 TI - Neonatal gastric pull up: reality or myth? AB - Between 1991 and 1998, 28 out of 356 neonates with esophageal atresia and tracheoesophageal fistula (EATEF) required esophageal substitution. As only 8 returned (28%) for the esophageal replacement after initial esophagostomy and gastrostomy at our center, we were prompted to offer single-stage esophageal replacement when the primary repair had either failed or was not found feasible. Twelve full-term neonates (mean birth weight 2.32 kg) with EATEF who underwent esophageal replacement by gastric pull-up between 1998 and 2000 were reviewed. The indications were: major leak after primary repair (n=9); pure EA (n=2); and EATEF with a very wide gap (n=1). The average ages at presentation and gastric pull-up were 6.0 and 8.5 days, respectively. The patients were evaluated for gastric transit by a colloid radiopharmaceutical, for duodenogastric reflux (DGR) by hepatic immunodiacetic acid (HIDA) scan, and for gastric clearance and transit by contrast studies. Three patients had minimal leaks from the neck site, all of which healed well. Follow-up with nuclear scans and contrast studies to evaluate gastric emptying revealed obstruction in 1 case and DGR in 25% of cases. There were 2 deaths (16%), 1 due to complex congenital cardiac disease and the other due to septicemia. In view of the acceptable morbidity, mortality, and functional outcome following gastric pull-up, we recommend this procedure if it becomes inevitable in the neonatal period. PMID- 12721738 TI - Rectum leiomyoma in a 10-month-old female. AB - An unexpected case of leiomyoma (LM) of the rectum in a 10-month-old female patient. The patient presented with a palpable mass and symptoms of intestinal obstruction, constipation and rectal discomfort. Rectal examination revealed a clearly visible mass. The treatment consisted of a surgical resection with wide margins. Pathology reported a leiomyoma. The patient was submitted to a careful clinical evaluation and a continuous follow up. PMID- 12721739 TI - Anorectal anomaly associated with caudal regression: late evaluation after posterior sagittal anorectoplasty. AB - The authors present a case of an anorectal anomaly associated with caudal regression syndrome, sirenomelia (mermaid syndrome), and a spectrum of mesodermal axial dysplasia. It was treated with a colostomy, posterior sagittal anorectoplasty, and closure of the colostomy. The patient is now 7-years-old, with fecal and urinary continence. Comments are made regarding the clinical and surgical findings, the follow-up evaluation, and anorectal manometry in this rare case. PMID- 12721740 TI - Congenital cystic mesoblastic nephroma. AB - With widespread use of antenatal ultrasound, an increasing number of urinary tract lesions are being detected, and the clinician is called upon to make management decisions. Parenchymal lesions of the kidney pose a special challenge because of the associated risk of the occurrence of malignancy. A case of an antenatally-detected solid/cystic lesion of the kidney diagnosed as a congenital cystic mesoblastic nephroma following nephrectomy is presented and the issues involved in therapeutic decision-making are discussed. PMID- 12721741 TI - Hepatoblastoma associated with Beckwith-Wiedemann syndrome and hemihypertrophy. AB - Both Beckwith-Wiedemann syndrome (BWS) and hemihypertrophy (HH) have been recognized to be overgrowth syndromes associated with an increased risk of cancer. We report an infant with hepatoblastoma associated with both BWS and HH in whom high serum alpha-fetoprotein (AFP) levels persisted even after complete tumor resection with no tumor recurrence. This phenomenon might be partly due to the nature of the proliferative disease. It is important to recognize that in some infants with BWS prolonged high serum AFP levels mimic the existence of a tumor, and that treatment should be based not only on AFP measurement, but also on repeated radiologic imaging. PMID- 12721742 TI - Mesenteric lipoblastoma in a 2-year-old child. AB - An usual case of mesenteric lipoblastoma in a 2-year-old child is presented, and the literature on the subject is reviewed. PMID- 12721743 TI - Adenomatous hyperplastic polyp of the gall bladder associated with cholelithiasis in a child. AB - Polyps of the gall bladder are uncommon conditions in children. We present a case report of a 14-year-old girl who had calculous cholecystitis and an adenomatous hyperplastic polyp of the gall bladder. She was treated by laparoscopic cholecystectomy. PMID- 12721744 TI - Strangulated left paraduodenal hernia in an infant. AB - Paraduodenal hernia is a rare cause of intestinal obstruction. It occurs typically in adults, but can also present in children. The symptoms include chronic abdominal pain and acute intestinal obstruction. Rapid evaluation must be followed by early surgery to avoid the morbidity and mortality associated with strangulation. PMID- 12721745 TI - Use of pyloric exclusion with a double jejunostomy in the treatment of a serious duodenal lesion in a child. AB - A 3-year-old male was beaten by his stepfather, resulting in a lesion of the third portion of the duodenum that was treated with an occlusive suture of the pyloric mucosa for pyloric exclusion and a gastric-jejunal anastomosis with a double jejunostomy for alimentation with a good result, no serious complications, and relatively a short hospitalization. This procedure may be an alternative for other similar cases. PMID- 12721746 TI - Buttressing the divided bladder neck by a rectus abdominis muscle flap to prevent urethral recanalisation in paediatric urinary incontinence. AB - Division of the bladder neck (BN) along with an augmentation cystoplasty and Mitrofanoff procedure was successfully employed in a group of patients with urinary incontinence. The BN division was buttressed with a unilateral vascularised rectus muscle flap, which effectively prevented urethral recanalisation without any cosmetic or functional disability. PMID- 12721747 TI - Port-related complications in children. PMID- 12721748 TI - Paediatric surgery in sub-Saharan Africa. PMID- 12721749 TI - Fetus in fetu. PMID- 12721750 TI - Limitations of oximeters. PMID- 12721751 TI - Thoughts on the implantation of prosthetic material at the esophageal hiatus in children. PMID- 12721753 TI - Prostatitis: ancient history and new horizons. PMID- 12721754 TI - Staurosporine-induced apoptosis is independent of p16 and p21 and achieved via arrest at G2/M and at G1 in U251MG human glioma cell line. AB - OBJECTIVE: The mechanisms involved in the cell cycle and cell death remain unresolved despite much investigation. Staurosporine induces cell death and G1 or G2/M arrest in a dose-dependent manner, but the mechanisms remain unknown. METHODS: In the present study an adenovirus vector expressing p16 or p21 genes in human glioma cell lines was used to examine cell cycle regulation and cell death induced by staurosporine. RESULTS: A low concentration (/=30 n M) induced G2/M arrest and finally induced apoptosis via a caspase-3 activated pathway from both the G2/M and G1 phases. However, pRb was dephosphorylated and cdc2 was inhibited at both the low and the high concentrations of staurosporine, indicating that the mechanisms of cell cycle regulation are not simply p53-Rb- or cdc2-dependent pathways. CONCLUSIONS: Forced G1 arrest by transfection with p16 or p21 genes did not alter the rate of staurosporine-induced cell death. This implies that an unknown pathway of apoptosis occurs from the G1 phase. PMID- 12721755 TI - Tamoxifen induces apoptosis in Fas+ tumor cells by upregulating the expression of Fas ligand. AB - PURPOSE: Tamoxifen (TAM), a nonsteroidal anticancer agent, is used in the treatment of breast cancer. In the current study, we investigated whether TAM induces apoptosis in tumor cells by altering the expression of Fas and Fas ligand (FasL). METHODS: Several tumor cell lines were used to test the ability of TAM to induce apoptosis, which was studied using the TUNEL assay. The effect of TAM on the expression of Fas and FasL was analyzed using a flow cytometer. RESULTS: TAM was found to suppress the growth of an estrogen receptor-positive human mammary tumor cell line (T-47D) by inducing apoptosis. Interestingly, TAM also induced apoptosis in an estrogen receptor-negative murine T cell lymphoma cell line, EL 4. The ability of TAM to induce apoptosis in T-47D and EL-4 tumor cells correlated with the increased expression of FasL but not Fas on the tumor cells. Similar to TAM, a metalloproteinase (MP) inhibitor, which is known to increase the expression of membrane-bound FasL, was found to induce apoptosis in both T 47D and EL-4 tumor cells by increasing the expression of FasL but not Fas. Furthermore, both TAM and the MP inhibitor failed to induce apoptosis in L1210 tumor cell lines that failed to express FasL. CONCLUSIONS: The current study demonstrates that TAM can induce apoptosis in Fas(+) tumor cells by upregulating FasL. PMID- 12721756 TI - The schedule-dependent effects of etoposide in leukaemic cell lines: a function of concentration and duration. AB - PURPOSE: Etoposide is a commonly used anticancer agent that is highly schedule dependent. The in vitro activity of etoposide (0-10 microM) was investigated in a panel of leukaemic cell lines. METHODS: Cells were cultured with etoposide in drug schedules of equal exposure duration (ED, durationxconcentration), and the effects of drug exposure on cell parameters, including cell cycle distribution, were assessed over an 8-day period. RESULTS: Proliferation assays indicated a concentration- and duration-dependent induction of cell death by etoposide in CEM and HL60 cells, and flow cytometric analysis indicated that this cell kill was by apoptosis. Efficacy was also dependent upon schedule, with more cell kill seen in schedules of longer duration. As an example, accumulative percent cell kill resulting from a continuous exposure to 0.05 microM etoposide was significantly greater than that in cultures involving either a 4-day exposure to 0.1 microM or a single-day exposure to 0.4 microM etoposide (193.4+/-15.9% vs 125.2+/-5.4% vs 42.3+/-5.9%, respectively; P<0.001 in all cases; equi-ED 0.4 microM.days). Efficacy was also dependent upon the ED of the schedule. At very low concentrations, the initial enhancement of cytotoxicity mediated by increasing duration would gradually and paradoxically be lost in the more protracted schedules (e.g. accumulative percent cell kill 66.4+/-7.4%, 158.3+/-12.0% and 40.1+/-6.0% with 100 n M for 2 days, 33 n M for 6 days and 25 n M for 8 days, respectively; P<0.001 in all cases; equi-ED 0.2 microM x days). CONCLUSIONS: Our results confirm the schedule-dependency of etoposide in vitro, highlighting the importance of total duration of drug exposure in determining cytotoxicity, and emphasizing the requirement to achieve a cytotoxic concentration in longer exposures. It is therefore crucial to ensure that etoposide regimens used clinically involve doses that are effectively cytotoxic. PMID- 12721757 TI - Safety and efficacy of the MDR inhibitor Incel (biricodar, VX-710) in combination with mitoxantrone and prednisone in hormone-refractory prostate cancer. AB - PURPOSE: VX-710 (biricodar, Incel) restores drug sensitivity to cells expressing P-glycoprotein (P-gp) and multidrug resistance-associated protein (MRP1). MRP1 is expressed in a high proportion of prostate tumors while P-gp expression is variable. Since mitoxantrone (M) and prednisone (P) are substrates for MDR transporters, we initiated a study to evaluate the safety, pharmacokinetics, and efficacy of VX-710 plus M/P in patients with hormone-refractory prostate cancer (HRPC). PATIENTS AND METHODS: Eligible patients had progressive HRPC (defined as new lesions, new disease-related pain, or 50% increase in PSA within 6 weeks of entry), testosterone <30 ng/ml, no prior chemotherapy, ECOG performance status of 0-3, and adequate organ function. Patients received VX-710 (120 mg/m(2) per h) as a 72-h continuous intravenous infusion with intravenous bolus mitoxantrone (12 mg/m(2)) administered 4 h after VX-710 was started and prednisone (5 mg twice daily) administered throughout the study treatment. Endpoints included serum PSA response, PSA response duration, time to PSA progression, pain reduction, and quality of life measures. RESULTS: Enrolled in the study were 40 patients and 184 courses of VX-710 plus M/P were administered. Intensive pharmacokinetics, which were performed on six patients who received one cycle of M/P alone, followed by VX-710 plus M/P for all other cycles, showed that VX-710 did not alter mitoxantrone clearance. VX-710 blood concentration at the time of mitoxantrone administration averaged 4.52 microg/ml. VX-710 plus M/P was well tolerated. Transient nausea/vomiting and mild neutropenia were the principal treatment toxicities. Five patients experienced an uncomplicated febrile neutropenic episode (12%), three had severe nausea/vomiting, and two experienced transient moderate to severe ataxia. Of the 40 patients, 12 (30%, 95% confidence interval 16-44%) had a reduction in PSA of >/=50% and 9 of the 12 patients (23% overall, 95% CI 10-35%) achieved a reduction in PSA of >/=80% that was sustained for the duration of treatment with M/P plus VX-710. The median time to PSA progression was 41 weeks (95% CI 34-68 weeks). Of the 40 patients, 15 completed treatment with stable disease and 13 had progressive disease with increasing serum PSA during study treatment. Median survival was 48 weeks for the intent-to-treat population of 40 patients. CONCLUSIONS: The addition of VX-710 to M/P therapy did not appear to increase the proportion of patients with significant serum PSA reductions compared to M/P alone. However, the duration of PSA response observed for the 12 PSA responders suggests that MDR inhibition may benefit some patients with HRPC. In addition to MRP1 or P-gp expression, other mechanisms of drug resistance are probably associated with the relative insensitivity of HRPC to cytotoxic therapy. PMID- 12721758 TI - The cardioprotector monoHER does not interfere with the pharmacokinetics or the metabolism of the cardiotoxic agent doxorubicin in mice. AB - PURPOSE: Monohydroxyethylrutoside (monoHER) has proved to be a good protector against doxorubicin-induced cardiotoxicity without interfering with the antitumor effect of doxorubicin. The aim of the present study was to determine whether there is a pharmacokinetic interaction between monoHER and doxorubicin which may be involved in monoHER cardioprotection. METHODS: Mice were treated with monoHER (500 mg x kg(-1) i.v.) alone, monoHER 5 min after doxorubicin (10 mg x kg(-1) i.v.), doxorubicin alone and doxorubicin 5 min after monoHER. The levels of monoHER and doxorubicin(ol) in plasma and heart tissue were measured by HPLC 24 h and 48 h after monoHER and doxorubicin administration, respectively. RESULTS: The areas under the concentration-time curves (AUCs) of monoHER and doxorubicin(ol) were not affected by the coadministered drug. No changes were observed in pharmacokinetic parameters such as initial and final half-lives, mean residence time, clearance and volume of distribution of monoHER and doxorubicin(ol) after single or combined administration. CONCLUSION: The cardioprotection of monoHER in mice is not caused by a pharmacokinetic interaction between monoHER and doxorubicin. PMID- 12721759 TI - High-dose methotrexate: on the relationship of methotrexate elimination time vs renal function and serum methotrexate levels in 1164 courses in 264 Swedish children with acute lymphoblastic leukaemia (ALL). AB - PURPOSE: The objectives of the present study were to determine the relationship between methotrexate (MTX) elimination time and various aspects of renal function and to evaluate the prognostic value of elevated serum MTX and creatinine for delayed MTX elimination. PATIENTS AND METHODS: The majority of the 264 children were being treated for ALL. According to the NOPHO-92 protocol, 5 or 8 g MTX/m(2) was administered over 24 h. Serum creatinine was assessed daily. In 11 patients from one centre, renal function was studied in more detail using serum cystatin C, iohexol clearance, and urinary albumin, IgG and protein HC. RESULTS: Increased serum creatinine correlated significantly with the elimination time of MTX, whereas no indications were found of tubular or barrier function damage. Of the 1164 courses, 44 had delayed elimination of MTX (>/=120 h). Serum MTX >150 microM at the end of infusion had a sensitivity of 0.27 and a specificity of 0.94 to predict delayed MTX elimination, and >/=50% increase in serum creatinine during the first treatment day (creatinine ratio) had a sensitivity of 0.32 and a specificity of 0.99. The corresponding risk ratios were 5 and 19 for MTX >150 micro M and creatinine ratio, respectively. In courses with a normal elimination time (<72 h), 99% of the courses had a rise in serum creatinine of less than 50%. CONCLUSIONS: Elevation of serum creatinine by more than 50% is a better predictor of delayed elimination than the level of serum MTX at the end of MTX infusion, especially if information on previous creatinine measurements is used to reduce the impact of an occasionally low serum creatinine value before the start of the MTX infusion. PMID- 12721760 TI - Circulating angiogenic growth factor levels in mice bearing human tumors using Luminex Multiplex technology. AB - Tumor angiogenesis is essential for tumor growth and metastasis formation. Luminex methodology was used to measure the levels of four angiogenic cytokines in cell culture medium and in the plasma of mice bearing human tumors. We obtained plasma and conditioned culture medium from 12 different human tumor cell lines. Tumor necrosis factor-alpha (TNF-alpha), basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), and transforming growth factor beta (TGF-beta) were determined by the Luminex FlowMetrix assay. VEGF, TNF-alpha, and bFGF were undetectable in non-tumor-bearing animals. HS746T gastric cancer and Caki-1 renal cell cancer cells in culture produced high levels of VEGF (1000 and 450 pg/10(6) cells, respectively). High levels of TGF-beta were produced by HS746T gastric carcinoma and Calu-6 non-small-cell lung carcinoma (3000 and 1000 pg/10(6) cells, respectively). Caki-1 renal cell carcinoma and Calu-6 non-small cell lung carcinoma cells in culture produced high levels of bFGF (42 and 10 pg/10(6) cells, respectively). Caki-1, SW2 SCLC, HCT-116 and HT-29 colon tumors produced high plasma levels of VEGF (200, 220, 42, and 151 pg/ml, respectively) and TGF-beta (31, 36, 45, 32 pg/ml, respectively). A positive linear correlation was seen between tumor volume and VEGF in SW2 (r=0.87) and Caki-1 (r=0.47) tumors, and a moderate correlation in HCT116 tumors (r=0.3). Angiogenic profiles in the plasma of nude mice bearing human tumors may be useful to identify appropriate biomarkers for antiangiogenic therapy, as diagnostic and prognostic tools, and to monitor the responses of individual tumors to antiangiogenic therapy. PMID- 12721761 TI - Gemcitabine pharmacokinetics and interaction with paclitaxel in patients with advanced non-small-cell lung cancer. AB - PURPOSE: Gemcitabine administered at a fixed dose rate of 10 mg/m(2) per min has been reported to achieve plasma steady-state concentrations ranging from 10 to 20 microM in patients with acute leukemia. These concentrations have been shown to saturate the intracellular accumulation of the active triphosphate metabolite. We designed this pharmacokinetic study to assess the ability of a fixed dose rate of gemcitabine to achieve the desired steady-state concentration in the absence and presence of paclitaxel in patients with solid tumors. PATIENTS AND METHODS: A group of 14 patients with advanced non-small-cell lung cancer received paclitaxel 110 mg/m(2) over 3 h on days 1 and 8 and gemcitabine 800 mg/m(2) over 80 min on days 1 and 8 every 21 days. Patients received gemcitabine alone on cycle (C) 1, day (D) 1. Pharmacokinetic samples were collected at 0, 15, 30, 45, 60 and 80 min during infusion and 0.25, 0.5, 1, 2, 4, 6, and 8 h after infusion on C1D1, C1D8, C2D1, C4D1 and C6D1. RESULTS: Of 13 patients included in the pharmacokinetic analysis, 61% achieved the desired steady-state concentration (C(ss)) with gemcitabine alone (C1D1), whereas only 0 to 45% of patients achieved the desired C(ss) with paclitaxel and gemcitabine, depending on the treatment cycle. Paclitaxel significantly decreased systemic clearance (Cl(T); P=0.012) and volume of distribution (V(d); P=0.050) and significantly increased C(ss) ( P=0.009). Gemcitabine plasma pharmacokinetic parameters demonstrated great interpatient variability in the absence of paclitaxel (C(ss) 30%, Cl(T) 30%, V(d) 55%). Interpatient and intrapatient variability in gemcitabine pharmacokinetics were not observed when gemcitabine was administered in combination with paclitaxel (P>0.05). CONCLUSIONS: Gemcitabine plasma pharmacokinetic parameters are significantly altered in the presence of paclitaxel. PMID- 12721762 TI - Cell cycle-dependent potentiation of cisplatin by UCN-01 in non-small-cell lung carcinoma. AB - PURPOSE: We evaluated the combination of UCN-01 plus cisplatin and sought to determine how the cell cycle effects of each agent affected the combined response. Cisplatin-induced DNA damage results in cell cycle arrest, primarily at the S and G(2) checkpoints, providing the opportunity for DNA damage repair prior to mitosis. Thus, strategies to enhance cisplatin cytotoxicity include attenuation of DNA damage-induced checkpoints. The cyclin-dependent kinase inhibitor 7-hydroxystaurosporine (UCN-01) can potentiate cisplatin activity, likely via abrogation of the S and G(2) checkpoints. UCN-01 has additional effects on cell cycling, including induction of an RB-associated G(1) arrest. METHODS: NSCLC cell lines A549 (wt p53, wt RB), Calu1 ( p53-null, wt RB) and H596 (mt p53, RB-null) were treated with UCN-01 and/or cisplatin with two-drug treatments delivered in alternate sequences. Effects of drug treatment on cell growth, cell cycling, apoptosis and levels and phosphorylation of cell cycle associated proteins were evaluated. The interaction between the two drugs was assessed using median effect analysis. RESULTS: When UCN-01 preceded cisplatin, growth inhibition was additive or less than additive, as assessed by median effect analysis. In contrast, when NSCLC cells were treated with cisplatin followed by UCN-01, the combination was synergistic. In this treatment sequence, a decrease in the proportion of cells at the G(2) checkpoint was confirmed by reduced expression of cyclins A and B and activation of Cdk1. Abrogation of the G(2) DNA damage checkpoint and apoptosis were prevalent only in cell populations treated with cisplatin followed by UCN-01 and was markedly enhanced in the cell lines with disrupted p53. CONCLUSIONS: These studies indicate that timing of drug administration strongly influences response to cisplatin plus UCN-01 in NSCLC cells, and this is related to the cell cycle-modulatory effects of these agents. Furthermore, this sequence combination was more effective in cell lines with dysfunctional p53. These findings support the hypothesis that checkpoint abrogation is the major mechanism of UCN-01-mediated potentiation of cisplatin cytotoxicity. PMID- 12721763 TI - Oxygen-15 positron-emission tomography for predicting selective delivery of a chemotherapeutic agent to hepatic cancers during angiotensin II-induced hypertension. AB - PURPOSE: Selective drug delivery is important for successful chemotherapy. In patients with hepatic cancer, we used (15)O-carbon dioxide (steady-state) and (15)O-water (dynamic) positron emission tomography (PET) to evaluate changes in blood flow induced by angiotensin (AT) II in the liver parenchyma, hepatic tumors, and spleen. METHODS: The study group comprised 13 patients, 6 with hepatic metastasis and 7 with hepatocellular carcinoma. PET was performed before and during AT II administration to induce hypertension. RESULTS: Steady-state PET images showed a preferential increase in blood flow to hepatic tumor compared with liver parenchyma during hypertension. In dynamic PET, tumor blood flow was maintained during induced hypertension while hepatic arterial blood flow, portal blood flow, total hepatic blood flow, and splenic blood flow decreased to 71.4% (not significant), 65% (P<0.01), 67.2% (P<0.001), and 72.3% (P<0.01) of the respective baseline values. CONCLUSION: AT II-induced hypertension produced a relative increase in tumor blood flow. Oxygen-15 PET should be useful for predicting preferential drug delivery to hepatic cancers. PMID- 12721764 TI - Bleomycin-mediated electrochemotherapy in mouse NR-S1 carcinoma. AB - PURPOSE: To determine whether or not low-voltage electrochemotherapy has cell killing effects. MATERIALS: Dorsally transplanted NR-S1 carcinomas in mice were stimulated with electric pulses (40 V/cm) after bleomycin (1 microg/g) had been injected around them. The tumors were fixed with a forceps electrode and electroporation was carried out three times a day for 4 days per week for 2 weeks. RESULTS: After 8 weeks of experimentation, the tumor had disappeared in four of the ten mice. The cell killing effects were mainly apoptosis and necrosis. CONCLUSION: Electroporation should be clinically introduced into the cosmetic and functional treatment of the head and neck region. Further investigation is also necessary to determine suitable carcinostatic agents and clarify the electric pulse conditions. PMID- 12721766 TI - Current therapies in Hodgkin's disease. AB - Advances in the treatment of Hodgkin's disease (HD) have resulted in cure rates of greater than 80%. This remarkable achievement has occurred in the past 50 years secondary to improvements in combination chemotherapy and radiotherapy. Over the last several decades, with the increase in long-term survivors of HD, it has become evident that cure is not the only issue, and late side-effects of treatment, including secondary malignancies and impaired fertility, are of major concern as well. As a result, attempts to improve response and survival rates by intensifying therapy must be countered against the potential for long-term toxicity. PMID- 12721767 TI - Inhibition of early 99mTc-MIBI uptake by Bcl-2 anti-apoptotic protein overexpression in untreated breast carcinoma. AB - Lack of technetium-99m methoxyisobutylisonitrile ((99m)Tc-MIBI) uptake is consistently reported to predict poor response to subsequent chemotherapy in a variety of human malignant tumours. Since (99m)Tc-MIBI accumulates within mitochondria, which also play a central role in apoptosis through the integration of death signals by Bcl-2 family members, we tested whether early (99m)Tc-MIBI uptake is affected by alterations of the apoptotic pathway. Forty-two breast cancer patients were intravenously injected with 740 MBq of (99m)Tc-MIBI and planar images were obtained 10 min post injection with the patients in the prone lateral position. Ten carcinomas failed to accumulate (99m)Tc-MIBI and could not be visualised on scintigraphic images despite being larger than 1.8 cm (MIBI negative). Thirty-two of the 42 breast carcinomas showed focal uptake of (99m)Tc MIBI (MIBI positive), and 10 min tumour-to-background ratios (T/B) varied between 1.14 and 6.93. The apoptotic index, the rate of proliferation, and the expression of the anti-apoptotic Bcl-2 protein and pro-apoptotic Bax protein were assessed in surgically excised tumours. All MIBI-negative carcinomas showed a dramatic and statistically significant reduction in the apoptotic index as compared with MIBI positive lesions (mean+/-SD, 0.14+/-0.15 vs 1.28+/-0.83, P<0.0001) independently of rate of proliferation, tumour size and P-glycoprotein expression. Significantly higher levels of Bcl-2 were also found in MIBI-negative as compared with MIBI-positive carcinomas. In MIBI-positive lesions, an inverse significant correlation was found between T/B ratios and Bcl-2 levels ( r=-0.50, P<0.01). Our findings indicate that early uptake of (99m)Tc-MIBI in breast carcinomas is affected by alterations of apoptotic pathway. High levels of Bcl-2, despite the stabilisation of mitochondrial membrane potentials, prevent accumulation of (99m)Tc-MIBI in tumour cells. In conclusion, absent or reduced early (99m)Tc-MIBI uptake in large tumours may indicate a Bcl-2-mediated resistance to chemo- and radiotherapy. PMID- 12721769 TI - Undergraduate teaching of nuclear medicine in European universities. AB - This paper provides an overview of the curricula of undergraduate training in nuclear medicine in 77 European medical departments and, for comparison, in nine departments outside Europe. The data show a high level of variation in the number of hours (0-62) devoted to nuclear medicine in the different departments. In most cases this teaching is integrated into one of the radiology or clinical modules, and in some cases also into training in clinical physiology. The paper discusses the differences in the particular approaches to nuclear medicine teaching. PMID- 12721768 TI - Combined low dose radio- and radioimmunotherapy of experimental HeLa Hep 2 tumours. AB - Radiation therapy of malignant tumours can be delivered by external beam radiation (RT) or radioimmunotherapy (RIT), using nuclides attached to monoclonal antibodies (mAbs). These treatment modalities have now been combined in order to investigate putative therapeutic advantages and elucidate the biological responses involved. Nude mice were transplanted subcutaneously on the back with human HeLa Hep2 tumour cells. RT (3x5 Gy) and/or 100 microg (131)I-labelled mAb H7, against placental alkaline phosphatase, or (131)I-labelled mAb TS1, against cytokeratin, was administered separately or in combination (specific activity of 120-200 MBq/mg antibody). Significant tumour growth retardation was observed both with RT alone and with RIT alone. Combining these regimens enhanced the therapeutic effects further, and a significant reduction in tumour volume could be demonstrated. The tumours were subjected to extensive histochemical and immunohistochemical investigations in order to elucidate changes in biology and histology within them. The following stainings were used: haematoxylin-eosin (morphology), Ki67 (proliferation), M30 (apoptosis), TUNEL (apoptosis) and endoglin (vascularisation). Tumours in the control group grew fast, with an average tumour doubling time of 9 days. These tumours contained large viable tumour cell masses displaying vast proliferation zones of Ki67-positive tumour cells, as well as necrotic regions and small amounts of connective tissue. Apoptotic cells could be identified both with M30 and TUNEL staining. When RT was applied, the growth rate was significantly reduced (doubling time 19 days) and typical alterations in morphology were seen, with a relative increase in connective tissue and a decrease in necrotic regions. Apoptotic cells were identified and a decrease in cell density was also observed. When RIT alone was applied, the growth parameters indicated a longer lasting growth reduction, especially when TS1 was used separately or in combination with H7. The histological appearances of these tumours were somewhat different from the RT treated tumours, with a larger portion of intratumoural cysts. These tumours also presented a reduced tumour cell density. Dramatic effects were observed when RT was combined with RIT, with a pronounced growth reduction seen in all combination treatment groups. Pronounced tumour volume reduction was also evident in both the RT + RIT ((131)I-TS1) group and RT + RIT ((131)I-TS1/(131)I-H7) group, and in some animals no tumour remained at all. The morphology of the tumour remnants at day 22 was chaotic with a drastically changed histology, with presence of abundant cysts, low fractions of Ki67-positive cells, reduction in cell density, increased amounts of connective tissue and a decrease in necrotic regions. Again, apoptotic cells could be identified, scattered throughout the viable regions. Combining RT and RIT seems to generate an efficient treatment with convincing and long-lasting tumour growth inhibition, which is reflected in a highly aberrant histology within the tumour. Results obtained in this study indicate that both necrosis and apoptosis may be involved in the process leading to this efficient therapy of epithelially derived tumours. PMID- 12721770 TI - A statistical investigation of amphiphilic properties of C-terminally anchored peptidases. AB - A number of DD-peptidases have been reported to interact with the membrane via C terminal amphiphilic alpha-helices, but experimental support for this rests with a few well-characterized cases. These show the C-terminal interactions of DD carboxypeptidases to involve high levels of membrane penetration, DD endopeptidases to involve membrane surface binding and class C penicillin-binding proteins to involve membrane binding with intermediate properties. Here, we have characterized C-terminal alpha-helices from each of these peptidase groups according to their amphiphilicity, as measured by mean , and the corresponding mean hydrophobicity, . Regression and statistical analyses showed these properties to exhibit parallel negative linear relationships, which resulted from the spatial ordering of alpha-helix amino acid residues. Taken with the results of compositional and graphical analyses, our results suggest that the use of C-terminal alpha-helices may be a universal feature of the membrane anchoring for each of these groups of DD-peptidases. Moreover, to accommodate differences between these mechanisms, each group of C-terminal alpha-helices optimizes its structural amphiphilicity and hydrophobicity to fulfil its individual membrane-anchoring function. Our results also show that each anchor type analysed requires a similar overall balance between amphiphilicity for membrane interaction, which we propose is necessary to stabilize their initial membrane associations. In addition, we present a methodology for the prediction of C-terminal alpha-helical anchors from the classes of DD-peptidases analysed, based on a parallel linear model. PMID- 12721773 TI - Drug formulations intended for the global market should be tested for stability under tropical climatic conditions. AB - RATIONALE OBJECTIVE: The quality of drugs imported into developing countries having a tropical climate may be adversely affected if their formulations have not been optimized for stability under these conditions. The present study investigated the influence of tropical climate conditions (class IV: 40 degrees C, 75% relative humidity) on the drug content, in vitro dissolution and oral bioavailability of different formulations of two essential drugs marketed in Tanzania: diclofenac sodium and ciprofloxacin tablets. METHODS: Before and after 3 and 6 months storage under class IV conditions the drug content and in vitro dissolution were evaluated using United States Pharmacopoeia (USP) 24 methods. Following a randomized four-period cross-over study, the pharmacokinetic parameters of drug formulations stored for 3 months under class IV conditions were compared with those stored at ambient conditions. RESULTS: Drug content and drug release from all tested ciprofloxacin formulations were within USP-24 requirements and remained stable during storage at simulated tropical conditions. Oral bioavailability was also not influenced by tropical conditions. The dissolution rate of two diclofenac formulations (Diclo 50 manufactured by Camden and Dicloflame 50 manufactured by Intas) reduced significantly during storage under class IV conditions. After oral administration Camden tablets stored for 3 months under class IV conditions showed a reduction in C(max) (90% CI of C(max) ratio: 0.59 - 0.76). This reduction was smaller than expected based on the in vitro tests. CONCLUSIONS: Some drug formulations imported into Tanzania are not optimized for stability in a tropical climate. Manufacturers and regulatory authorities should pay more attention to the WHO recommendations for testing the stability of drugs under tropical climate conditions. Efforts should be made to improve the in vitro tests to better predict the bioavailability. PMID- 12721772 TI - Validation of laser Doppler flowmetry coupled with intra-dermal injection for investigating effects of vasoactive agents on the skin microcirculation in man. AB - OBJECTIVE: To determine the reproducibility of laser Doppler flowmetry coupled with intra-dermal saline delivery. METHODS: Delivery of saline was judged visually by two operators ( n=100), using a graduated syringe (Becton-Dickinson), by expelling saline onto a weighing boat. Volume was assessed by weight. Skin blood flow following intra-dermal injection of saline was assessed in 18 healthy volunteers; 10 attended twice to assess between-day reproducibility, and 8 attended once to assess between-site reproducibility. Results are expressed as mean value+/-SEM and 95% confidence interval for mean differences. RESULTS: There was no difference between operators in mean injection weight, both weights being 10.3+/-0.1 mg (mean difference 0.08, 95% confidence interval, CI -0.23 to 0.39 mg; n=100, P=0.9). Intra-dermal saline caused a nine-fold increase in blood flow (0.03+/-0.003 to 0.27+/-0.02 perfusion units, PU; n=18, P<0.001). This response had a rapid onset, with the maximal effect seen at 4 min and a duration of greater than 30 min. There was no difference in the magnitude of the response between the dominant and non-dominant arms, AUC was 2.9+/-0.4 and 2.9+/-0.4, respectively (mean difference -0.05, 95% CI -0.8 to 0.73 PU; n=18, P=0.93). However, there was a trend towards differences between study visits 1 and 2: AUC was 3.2+/-0.6 and 2.0+/-0.5, respectively (mean difference 1.2, 95% CI -0.03 to 2.43 PU; n=10, P=0.7). There was no difference in the magnitude of responses between different sites on the forearm ( n=64, P=0.6). CONCLUSIONS: These studies demonstrate that the technique of laser Doppler flowmetry coupled with intra dermal injection is a safe, well-tolerated technique with good reproducibility. A trend towards reduced between-day reproducibility emphasizes the importance of vehicle control sites when investigating the effects of vasoactive compounds. This technique provides a reliable method for the intra-dermal delivery of drugs, despite the direct effect of injection of saline on blood flow. PMID- 12721775 TI - Synchrotron X-ray studies of the keel of the short-spined sea urchin Lytechinus variegatus: absorption microtomography (microCT) and small beam diffraction mapping. AB - In sea urchin teeth, the keel plays an important structural role, and this paper reports results of microstructural characterization of the keel of Lytechinus variegatus using two noninvasive synchrotron x-ray techniques: x-ray absorption microtomography (microCT) and x-ray diffraction mapping. MicroCT with 14 keV x rays mapped the spatial distribution of mineral at the 1.3 microm level in a millimeter-sized fragment of a mature portion of the keel. Two rows of low absorption channels (i.e., primary channels) slightly less than 10 microm in diameter were found running linearly from the flange to the base of the keel and parallel to its sides. The primary channels paralleled the oral edge of the keel, and the microCT slices revealed a planar secondary channel leading from each primary channel to the side of the keel. The primary and secondary channels were more or less coplanar and may correspond to the soft tissue between plates of the carinar process. Transmission x-ray diffraction with 80.8 keV x-rays and a 0.1 mm beam mapped the distribution of calcite crystal orientations and the composition Ca(1-x)Mg(x)CO(3) of the calcite. Unlike the variable Mg concentration and highly curved prisms found in the keel of Paracentrotus lividus, a constant Mg content (x = 0.13) and relatively little prism curvature was found in the keel of Lytechinus variegatus. PMID- 12721774 TI - Antidepressant drug use in general practice: inter-practice variation and association with practice characteristics. AB - OBJECTIVE: The use of antidepressants (ADs) has escalated and prompted considerable debate. Many depressed patients go unrecognised or under-treated and the area of indication of the new ADs is widening. The aim of this study was to analyse (i). the variation in general practitioners' prescribing of ADs by comparing with prescribing of other drug groups and (ii). whether the general prescribing behaviour, practice activity and demography are associated with the AD prescribing. METHODS: Analysis of AD prescribing patterns among 174 general practices (93.5%) in the County of Funen, Denmark. Age- and sex-standardised 1 year incidences and prevalences of AD prescribing for patients listed were calculated using individual prescription data from Odense University Pharmacoepidemiologic Database. Data about health services and practice demography were obtained from the Health Insurance Register. The variation in AD 1-year prevalence was compared with other drug groups by a variation index (90%/10% percentile). Univariate linear regression analysis was used to examine associations between practice characteristics and prescribing. RESULTS: The 1 year prevalence of AD prescribing varied sixfold, no more than the prevalence of five other drug groups. Practices with high yearly: general prescribing prevalence, mean number of drugs per medicated patient, number of surgery consultations/100 patients and counsellings/100 surgery consultations showed the highest yearly prevalence of AD prescribing. Single-handed practices had higher AD prescribing rates than partnerships. The relative use of selective serotonin re-uptake inhibitors and other new ADs showed only little variation (10% and 90% percentiles as close as 66-86%), but practices with high 1-year prevalence and incidence most often chose the new ADs. CONCLUSION: Analysis of inter-practice variation showed no extraordinary quality problems with regard to AD prescribing, but does not exclude that there might be problems. The general prescribing pattern of the general practitioners seems essential to their attitude to AD prescribing. The relationship between counselling and prescribing was a feature specific to ADs and deserves further investigation. Quality indicators are needed to understand differences in AD prescribing, and studies based on prescription data have to be supplemented with individual clinical data. PMID- 12721777 TI - Driving ability after acute and sub-chronic administration of levocetirizine and diphenhydramine: a randomized, double-blind, placebo-controlled trial. AB - RATIONALE: Sedation following antihistamine use poses a danger to ambulant patients involved in daily activities such as driving. OBJECTIVE: To investigate effects of levocetirizine (5 mg), diphenhydramine (50 mg), and placebo on driving ability during normal traffic. METHODS: Forty-eight healthy volunteers participated in a double-blind, placebo-controlled, randomized clinical trial. Treatments were administrated on days 1, 2, 3 and 4, exactly 1.5 h before the start of the standardized driving test (performed on day 1 and day 4). In the standardized driving test, subjects were instructed to drive with a steady lateral position, while maintaining a constant speed (95 km/h). Primary parameter was the standard deviation of lateral position (SDLP; cm). Statistical analyses were performed separately for day 1 and day 4, using analysis of variance and an equivalence test. Equivalence to placebo was evidenced if the 95% confidence interval lay between -2.6 cm and +2.6 cm. RESULTS: SDLP after levocetirizine was equivalent to placebo on both day 1 (-0.66 cm; +1.12 cm) and day 4 (-0.37 cm; +1.28 cm). In contrast, SDLP after diphenhydramine differed significantly from placebo on both day 1 ( P<0.0001) and day 4 ( P<0.0003). On day 1, the 95% confidence interval of diphenhydramine (+1.85 cm; +3.63 cm) was partially above the upper equivalence limit (+2.6 cm), indicating clinically relevant driving impairment. On day 4, however, the 95% confidence interval of diphenhydramine (+0.74 cm; +2.38 cm) was contained within the acceptance range. CONCLUSION: In contrast to diphenhydramine, driving performance was not significantly affected while using 5 mg levocetirizine once daily. PMID- 12721776 TI - Blockade of serotonin 5-HT1B and 5-HT2A receptors suppresses the induction of locomotor activity by 5-HT reuptake inhibitors, citalopram and fluvoxamine, in NMRI mice exposed to a novel environment: a comparison to other 5-HT receptor subtypes. AB - RATIONALE: Though 5-HT plays an important role in the modulation of motor function, which is perturbed in depressive states, little is known concerning the influence of serotonin reuptake inhibitors (SSRIs) on locomotor activity (LA). Recently, we demonstrated that SSRIs, such as citalopram, enhance LA in mice exposed to a novel environment. OBJECTIVES: This study examined the role of multiple classes of 5-HT receptor in citalopram-induced LA. METHODS: The most selective antagonists currently available were used. RESULTS: Citalopram-induced LA was dose-dependently attenuated by the 5-HT1B/1D receptor antagonists, S18127, GR125,743 and GR127,935, and by the selective 5-HT1B antagonist, SB224,289, but unaffected by the selective 5-HT1A antagonist, WAY100,635. The selective antagonists at 5-HT2A receptors, MDL100,907 and SR46,349 also dose-dependently attenuated induction of locomotion by citalopram, whereas the 5-HT2B antagonist, SB204,741, and the 5-HT2B/2C antagonist, SB206,553 were ineffective. Further, the selective 5-HT2C antagonist, SB242,084, potentiated the response to citalopram. Selective antagonists at 5-HT3 (ondansetron), 5-HT4 (GR125,487), 5-HT6 (SB271,046) and 5-HT7 (SB269,970) receptors did not significantly modify the action of citalopram. Underpinning these findings, SB224,289, GR125,743, MDL100,907 and SR46,349 likewise attenuated induction of locomotion by a further SSRI, fluvoxamine. CONCLUSIONS: The locomotor response to SSRIs of mice exposed to a novel environment is mediated via 5-HT1B and 5-HT2A receptors. In view of the importance of motor function to the etiology and treatment of depression, the significance of these observations to the clinical actions of SSRIs will be of interest to elucidate. PMID- 12721778 TI - The reinstatement model and relapse prevention: a clinical perspective. AB - OBJECTIVES: This commentary assesses the degree to which the reinstatement model is homologous to the human experience of relapse. RESULTS: A review of the literature suggests that the relationship is less clear than is often assumed, largely due to a lack of prospective data on the precipitants and process of relapse (especially relapse to heroin or cocaine abuse). However, reinstatement does not need to resemble relapse to have immediate clinical value; predictive validity as a medication screen would be sufficient. Whether the model has predictive validity is unknown, because, to date, very few clinical trials have tested medications that are effective in the reinstatement model, and even fewer have used designs comparable to those of reinstatement experiments. A clinical trial comparable to a reinstatement experiment would enroll participants who are already abstinent, and its main outcome measure would be propensity to undergo a specific type of relapse (e.g., relapse induced by stress or cues). CONCLUSIONS: Until clinical and preclinical work are more comparable, criticisms of the reinstatement model's presumed shortcomings are premature. PMID- 12721779 TI - Reduced ethanol-induced conditioned taste aversion and conditioned place preference in GIRK2 null mutant mice. AB - RATIONALE: Previous studies have shown that GIRK2 channel function is enhanced by ethanol and that GIRK2 null mutant mice are less sensitive to some of ethanol's effects, including anxiolysis, habituated locomotor stimulation, and acute handling-induced convulsions than wild types. Under some conditions, GIRK2 knockout mice consume more ethanol than wild types, but it is unclear whether they do so because they are more sensitive to ethanol's rewarding effects or less sensitive to its aversive effects. OBJECTIVE: To further assess the role of GIRK2 in ethanol action, GIRK2 null mutant and wild type mice were tested in conditioning models that measure the motivational effects of ethanol. METHOD: In a conditioned taste aversion (CTA) procedure, knockout and wild type mice were given ethanol (0.0, 2.0, 2.5, or 3.5 g/kg, IP) following 1-h access to saccharin every 48 h over a 10 day period. In a conditioned place preference (CPP) procedure, knockout and wild type mice were given ethanol (2.0 or 3.0 g/kg, IP) paired with one stimulus (grid or hole floor) and saline paired with the other. After four 5-min trials with each stimulus, a 60-min choice test was done. RESULTS: The results demonstrated a genotypic difference in both paradigms. In CTA, there was no difference between genotypes at 0.0 or 3.5 g/kg ethanol, but at the 2.0 and 2.5 g/kg doses, wild types developed a stronger aversion to saccharin than knockouts. In CPP, wild types developed place preference, but knockouts did not. CONCLUSIONS: These studies show that GIRK2 deletion reduced ethanol's impact in tasks that are commonly used to index the drug's rewarding and aversive effects. These findings could reflect either a learning/memory deficit or decreased sensitivity to ethanol's motivational effects in null mutant mice. The latter interpretation is more consistent with previous data showing that knockout mice consume higher doses of ethanol than wild type mice. PMID- 12721780 TI - Response to alcohol in females with a paternal history of alcoholism. AB - RATIONALE: Several studies have demonstrated that males with a family history of alcoholism (FHP) show less of a response to alcohol (e.g. lower ratings of intoxication) than males without a family history of alcoholism (FHN). The purpose of this pilot study was to determine if FHP females also showed a reduced sensitivity to alcohol compared to FHN females. OBJECTIVES: To determine if FHP females (n=16) were less sensitive to the subjective effects and performance impairing effects of alcohol compared to FHN females (n=16). METHODS: The effects of placebo and alcohol (0.25, 0.50, 0.75 g/kg, based on total body water) were evaluated using a double-blind, placebo-controlled outpatient design. Drug effects were assessed using performance tasks, observer ratings of drug effect and subjective ratings of drug effect. RESULTS: There were no differences in breath alcohol levels between FHN and FHP women. FHP women were less impaired by alcohol than FHN women, as shown by DSST scores and observer-ratings. However, FHP women were more impaired on the Digit Recall task after alcohol than FHN women and they tended to have higher ratings of "Good Drug Effect," "Drug Liking" and "Willingness to Take Again." Of note, FHP women reported more dysphoric mood than FHN women in the absence of alcohol administration. CONCLUSIONS: The results of the present study suggest that FHP women may have a reduced response to alcohol on some measures, but FHP women report greater positive effects on other measures. Overall, the differences between FHP and FHN women are subtle compared to the previous studies demonstrating a reduced response to alcohol in FHP men. PMID- 12721781 TI - Beyond the question of placebo controls: ethical issues in psychopharmacological drug studies. AB - RATIONALE: There is a broad range of complex ethical issues in the conduct of psychopharmacological drug studies that go beyond the question of the ethics of placebo controls. However, our empirical knowledge with respect to these issues is very limited. This review, although not exhaustive, highlights an array of ethical issues that arose from discussions within the NIMH Human Subjects Research Council Workgroup. OBJECTIVES: To delineate issues in psychopharmacological drug studies that require debate and would benefit from research leading to the development of empirically-supported guidelines. METHODS: Information included in this report was drawn from the first author's participation as chair of the NIMH Human Subjects Research Council Workgroup, guidelines for the ethical conduct of research proposed by professional organizations to which the first and third author belong, and relevant research literature. RESULTS: We have focused on general issues relating to informed consent, research with special populations, and long-term treatment studies. Additionally, we raise issues relevant to large research-oriented institutions. CONCLUSIONS: The essential ethical challenge in psychopharmacological trials is to balance risks and benefits in the context of the needs and capacities of individual research subjects. The IRB system must become evidence-based and not rely on unproven assumptions. Specific research studies should be undertaken to address many of the issues of informed consent and research ethics postulated in this paper. PMID- 12721783 TI - Medical-record review of potential donor pool in the Czech Republic suggests a possible increase to more than double the number of donors. AB - The objective of this study was the investigation of the maximal potential donor pool and causes of non-realized organ donation. On-site retrospective medical record review was performed for all cases of patient death ( n=1608) occurring in 1999 at 34 (83% of all) intensive care units (ICUs) in the region of the transplant center in Prague, Czech Republic. Two hundred and eighty-eight (18%) patients died with clinical signs of brain death. ICU physicians considered 111 of them as being potential donors at the time; 63 became donors and 48 did not. The remaining 177 patients with clinical signs of brain death were, in retrospect, assessed as being unsuitable ( n=105) or suitable ( n=72) for donation. The maximal potential donor rate was 55.7 per million population (pmp), with a more conservative estimate of 37.4 pmp. The actual donor rate was 18.1 pmp. For the maximal level to be approached, further educational efforts are warranted, targeting ICU physicians and concentrating on the identification of potential donors. PMID- 12721784 TI - Life-history parameters of a wild group of West African patas monkeys (Erythrocebus patas patas). AB - Based on long-term, although intermittent, observations (2 years 4 months of 14 years), we present data on birth seasonality, age at first birth, interbirth intervals, mortality rates, age at first emigration, and population change of a wild population of West African patas monkeys ( Etythrocebus patas patas) in northern Cameroon. Birth season was from the end of December until the middle of February, corresponding to the mid-dry season. In spite of large body size, the patas females had the earliest age at first birth (36.5 monthsold) and the shortest interbirth intervals (12 months) compared to the closely related wild forest guenons. Age at first emigration of the males was considered to occur between 2.5 and 4.5 years. The group size of the focal group drastically decreased between 1984 and 1987, and steadily increased until 1994, then decreased again in 1997. The neighboring group also showed a similar trend in group size. The population decreases were likely to be caused by drought over 3 years. Annual crude adult mortality rate was 4% during population increase periods (PIP) between 1987 and 1994. It rose to 22% during all the periods (AP), including drought over 3 years. Despite their smaller body size, the rate of the wild forest guenons ( Cercopithecus mitis) (4%) was the same and much lower than those of the patas during PIP and AP, respectively. The annual average juvenile mortality rate was 13% during PIP and it also rose to 37% during AP. That of wild forest guenons ( C. ascanius) (10-12%) was a little lower and much lower than those of the patas during PIP and AP, respectively. These findings were consistent with Charnov's theoretical model of mammalian life-history evolution in that patas with high adult and juvenile mortality showed early and frequent reproduction in spite of large body size. Charnov also considered high adult mortality as a selective force and high juvenile mortality as a density-dependent consequence of high fecundity. Our results support the former but not the latter research findings. PMID- 12721782 TI - Impulse activity of midbrain dopamine neurons modulates drug-seeking behavior. AB - RATIONALE: Withdrawal from non-contingent exposure to psychostimulants increases the activity of midbrain dopamine cells and impairs the function of impulse regulating dopamine autoreceptors. It is unclear whether these neuroadaptations play an important role in withdrawal-associated drug seeking. OBJECTIVES: We determined whether cocaine self-administration modifies the impulse activity of midbrain dopamine neurons and dopamine autoreceptor function, and whether experimentally induced reduction in dopamine cell activity (by autoreceptor activation) could influence drug-seeking behavior. METHODS: Animals were trained to self-administer saline or cocaine (500 micro g/kg per infusion) for 7 days. At different withdrawal periods, we used single-unit extracellular recordings to measure impulse activity of dopamine cells and administered the D2/D3 dopamine receptor agonist quinpirole to determine autoreceptor sensitivity. In a separate set of experiments, we determined the effects of autoreceptor-selective doses of quinpirole on drug-seeking behavior (non-reinforced responding in the absence of cocaine) during an extinction/reinstatement task. RESULTS: Cocaine self administration induced a short-lived increase in the mean firing rate and bursting activity of midbrain dopamine cells. This effect was greatest at early withdrawal and was paralleled by decreased ability of quinpirole to inhibit dopamine cell firing rate and drug-seeking behavior. Changes in dopamine cell activity dissipated over time; at late withdrawal, when both impulse activity and autoreceptor sensitivity returned to control values, quinpirole dramatically decreased drug-seeking behavior. CONCLUSIONS: These results show that inhibiting dopamine cell impulse activity, by activation of dopamine autoreceptors, reduces drug-seeking behavior. This suggests that the impulse activity of midbrain dopamine cells could be an important factor contributing to relapse. PMID- 12721785 TI - Biosynthesis of cytokinins. AB - Cytokinins are adenine derivatives with an isoprenoid side chain and play an essential role in plant development. Plant isopentenyltransferases that catalyze the first and rate-limiting steps of cytokinin biosynthesis have recently been identified. Unlike bacterial enzymes, which catalyze the transfer of the isopentenyl moiety from dimethylallyldiphosphate (DMAPP) to the N(6) position of adenosine 5'-monophosphate (AMP), plant enzymes catalyze the transfer of the isopentenyl moiety from DMAPP preferentially to ATP and to ADP. The isopentenylated side chain is hydroxylated to form zeatin-type cytokinins. An alternative pathway, in which a hydroxylated side chain is directly added to the N(6) position of the adenine moiety, has also been suggested. PMID- 12721786 TI - Structure and function of cytokinin oxidase/dehydrogenase genes of maize, rice, Arabidopsis and other species. AB - Cytokinin oxidases/dehydrogenases (CKX) catalyze the irreversible degradation of the cytokinins isopentenyladenine, zeatin, and their ribosides in a single enzymatic step by oxidative side chain cleavage. To date the sequences of 17 fully annotated CKX genes are known, including two prokaryotic genes. The CKX gene families of Arabidopsis thaliana and rice comprise seven and at least ten members, respectively. The main features of CKX genes and proteins are summarized in this review. Individual proteins differ in their catalytic properties, their subcellular localization and their expression domains. The evolutionary development of cytokinin-catabolizing gene families and the individual properties of their members indicate an important role for the fine-tuned control of catabolism to assure proper regulation of cytokinin functions. The use of CKX genes as a tool in studies of cytokinin biology and biotechnological applications is discussed. PMID- 12721788 TI - Task preparation in macaque monkeys ( Macaca mulatta). AB - We investigated whether macaque monkeys possess the ability to prepare abstract tasks in advance. We trained two monkeys to use different stimulus-response (S-R) mappings. On each trial, monkeys were first informed with a visual cue which of two S-R mapping to use. Following a delay, a visual target was presented to which they would respond with a left or right button-press. We manipulated delay time between cue and target and found that performance was faster and more accurate with longer delays, suggesting that monkeys used the delay time to prepare each task in advance. PMID- 12721789 TI - Catalog of 680 variations among eight cytochrome p450 ( CYP) genes, nine esterase genes, and two other genes in the Japanese population. AB - We screened DNAs from 48 Japanese individuals for single-nucleotide polymorphisms (SNPs) in eight cytochrome p450 ( CYP) genes, nine esterase genes, and two other genes by directly sequencing the relevant genomic regions in their entirety except for repetitive elements. This approach identified 607 SNPs and 73 insertion/deletion polymorphisms among the 19 genes examined. Of the 607 SNPs, 284 were identified in CYP genes, 302 in esterase genes, and 21 in the other two genes ( GGT1, and TGM1); overall, 37 SNPs were located in 5' flanking regions, 496 in introns, 55 in exons, and 19 in 3' flanking regions. These variants should contribute to studies designed to investigate possible correlations between genotypes and phenotypes of disease susceptibility or responsiveness to drug therapy. PMID- 12721790 TI - Association of natural tooth loss with genetic variation at the human matrix Gla protein locus in elderly women. AB - Natural tooth loss represents a major medical issue within the elderly population, since it impairs masticatory function critical for oral intake of essential nutrition. Contribution of genetic factors has been implicated in the determination of natural tooth loss; degree of reduction in number of natural teeth remaining intact (NTI) varies among individuals; thus, heterogeneity in NTI might reflect genetic variation within the population. One candidate gene, the matrix Gla protein gene (MGP), has been implicated in the pathogenesis of bone loss through a repression of bone/tooth formation. We have investigated a possible association between the CA repeat polymorphism at the human MGP gene locus and the NTI in 458 elderly Japanese women. In 916 chromosomes tested, ten alleles of the polymorphic nucleotide repeat were observed (designated A1-A10), among which five alleles were regarded as major alleles to be tested for the association. Twenty-seven women who possessed an A6 allele (164 bp) had significantly higher NTI than the remaining participants (n=431), who did not carry an allele of that size (mean: 10.0 teeth vs 5.6 teeth; P=0.007, Mann Whitney test). An eight-year longitudinal follow-up study of NTI suggested that the genetic variations at the MGP locus did not affect the rate of tooth loss in the elderly period. These results suggest that genetic variation at the MGP gene locus is associated with some determinants for tooth loss in elderly women. PMID- 12721791 TI - GPC5 is a possible target for the 13q31-q32 amplification detected in lymphoma cell lines. AB - Comparative genomic hybridization (CGH) analyses have detected gains of copy number on 13q, especially at 13q31-q32, in cell lines and primary cases of various types of lymphoma. Since amplification of chromosomal DNA is one of the mechanisms that can activate tumor-associated genes, and because 13q amplification had been reported in various other types of tumors as well, we attempted to define by fluorescence in situ hybridization (FISH) a common region at 13q31-q32 in which to explore genes that might be targets for the amplification events. Although the commonly amplified region we defined was relatively large (approximately 4 Mb), only one true gene, GPC5, was found there. GPC5 was over-expressed in lymphoma cell lines that had shown amplification, in comparison with those that had not. Our findings suggest that GPC5 is a likely target for amplification, and that over-expression of this gene may contribute to development and/or progression of lymphomas and other tumors. PMID- 12721792 TI - Virioplankton community structure along a salinity gradient in a solar saltern. AB - The virioplankton community structure along a salinity gradient from near seawater (40 per thousand ) to saturated sodium chloride brine (370 per thousand ) in a solar saltern was investigated by pulsed-field gel electrophoresis. Viral populations with genome sizes varying from 10 kb to 533 kb were detected. The viral community structure changed along the salinity gradient. Cluster analysis of the viral genome-banding pattern resulted in two main clusters. The virioplankton diversity within the samples with salinity from 40 per thousand to 150 per thousand was on the same cluster of a cladogram. The other group consisted of virioplankton from samples with salinity above 220 per thousand. The virioplankton diversity in the different samples was calculated using the Shannon index. The diversity index demonstrated an increase in diversity in the samples along the gradient from 40 per thousand to 150 per thousand salinity, followed by a decrease in the diversity index along the rest of the salinity gradient. These results demonstrate how viral diversity changes from habitats that are considered one of the most common (seawater) to habitats that are extreme in salt concentrations (saturated sodium brine). The diversity index was highest in the environments that lie in between the most extreme and the most common. PMID- 12721793 TI - Tissue distribution of the Ankara strain of vaccinia virus (MVA) after mucosal or systemic administration. AB - MVA is a candidate vector for vaccination against pathogens and tumors. Little is known about its behaviour in mucosal tissues. We have investigated the fate and biosafety of MVA, when inoculated by different routes in C57BL/6 mice. Intranasal inoculation targeted the virus to the nasal associated lymphoid tissue and the lungs, whereas systemic inoculation led to distribution of MVA in almost all lymphoid organs, lungs and ovaries. Intravaginal, intrarectal and intragastric inoculations failed to induce efficient infection. After 48 h no virus was detectable any more in the organs analyzed. Upon intranasal inoculation, no inflammatory reactions were detected in the central nervous system as well as the upper and lower airways. These results show the tropism of MVA and indicate that high doses of recombinant MVA are safe when nasally administered, a vaccination route known to elicit strong cellular and humoral immune responses in the female genital tract. PMID- 12721795 TI - Genetic and antigenic analysis of two recently circulating genotypes of type A foot-and-mouth disease virus in India: evidence for positive selection in the capsid-coding genes. AB - We have analyzed isolates of two recently circulating genotypes (genotypes VI and VII) of type A foot-and-mouth disease virus (FMDV) from India. Maximum-likelihood models provided support for the presence of positively selected sites in the capsid-coding (P1) region. Positive selection was detected at a number of amino acid positions behind a background of strong purifying selection. Among the positively selected sites, four were identified at known critical antigenic residues (VP2 79, VP3 59 and 70 and, VP1 83), suggesting that FMDVs are under pressure from the immune system. Two residues (VP2 134 and VP3 59) that are part of the heparan sulfate-binding pocket in subtype A22 FMDV are also inferred to be under positive selection. Antigenic divergence was observed between and within the genotypes in neutralization tests with sera raised against the representative isolates from genotypes VI and VII. The two vaccine strains showed one-way antigenic relationships (r value) of <0.2 with 64% of the isolates, whereas, with genotypes VI and VII an r value of >0.4 was observed with 24% and 64% of the isolates, respectively. No correlation could be deduced from the amino acid substitutions at specific critical residues and lower r values in the field isolates. PMID- 12721794 TI - Persisting TT virus (TTV) genogroup 1 variants in renal transplant recipients. AB - TT virus (TTV) genogroup 1 infection has an increased prevalence in solid organ transplant recipients. In this study, the presence of TTV in renal transplant recipients was examined by two PCR methods, one capable of detecting most TTV genotypes (UTR-PCR), the other specific to genogroup 1 (N22-PCR). The N22-PCR detected TTV in 57% (53/92) of the renal transplant patients and in 20% (13/66) of the healthy individuals, while the prevalence of TTV with the UTR-PCR was above 90% in both the control and the patient groups. The N22-PCR was used in longitudinal studies of 31 renal transplant recipients, these PCR products were sequenced and aligned. TTV status was not associated with the patients' age at transplantation, male to female ratio and the time lag between kidney transplantation and the TTV test. During the follow-up consistent TTV status was found in 26 patients, while two initially TTV positive patients converted to negative and three initially negative patients converted to positive. The TTV variants varied among the tested patients, but were the same in the consecutive samples of each patient, indicating that TTV infection was persistent in renal transplant recipients and novel infection occurred rarely in the post-transplant period. PMID- 12721796 TI - Kaposi's sarcoma-associated herpesvirus (KSHV)-encoded vMIP-I and vMIP-II induce signal transduction and chemotaxis in monocytic cells. AB - Kaposi's sarcoma-associated herpesvirus (KSHV)/ Human herpesvirus 8 encodes three chemokines, which are called viral macrophage inflammatory protein (vMIP)-I, -II, and -III. Here, we expressed the KSHV vMIP-I and vMIP-II proteins and analyzed their biological functions. Both vMIP-I and vMIP-II had an apparent molecular mass of 7.8 kDa and were localized to the cytoplasm in a body cavity-based lymphoma cell line BC-3, stimulated with phorbol ester. We next treated a human monocytic leukemia cell line, THP-1, with purified recombinant vMIP-I and vMIP II, or vMIP-I and vMIP-II fused with alkaline phosphatase to study Ca(2+) signalling and in vitro chemotaxis in response to these proteins. Calcium mobilization was induced by both vMIP-I and vMIP-II. Furthermore, vMIP-I and vMIP II induced Ca(2+) mobilization in K562 cells expressing the CC chemokine receptor 5 (CCR5), suggesting that both may be agonistic for CCR5. Additionally, vMIP-I induced Ca(2+) mobilization through the intermediary of CCR8. These viral MIPs were also capable of chemotactically activating the THP-1 cells. These results imply that vMIP-I and vMIP-II may play important roles in the propagation of KS and primary effusion lymphoma by inducing the chemotaxis of CCR5-expressing monocytes. PMID- 12721797 TI - Rapid serological profiling by enzyme-linked immunosorbent assay and its use as an epidemiological indicator of foot-and-mouth disease viral activity. AB - Frequency distribution of reactivity levels of foot-and-mouth disease infection specific antibodies in livestock populations was analysed. Specific antibody responses against non-capsid polyprotein 3ABC were assessed through a highly sensitive indirect enzyme-linked immonosorbent assay (I-ELISA 3ABC). A graphic display of data was designed based on three negative and three positive categories to illustrate reactivity patterns. The resulting patterns were correlated to the epidemiological status. On this basis, results of over 100,000 sera derived from cattle populations in regions with various well-documented epidemiological situations were compiled and are exemplified in this paper.Distinct distributions of antibody reactivity patterns reflecting the various epidemiological situations were attained. Whereas non-affected areas presented a rather homogenous negative pattern with very limited test-positive reactions, affected regions revealed quite heterogeneous profiles, including positive and negative categories, with distributions that varied according to the region. The use of graphic prints encompassing I-ELISA 3ABC antibody profile responses constituted an adequate epidemiological indicator of the risk of foot and-mouth disease viral activity, providing immediate visualization for a rapid inference of the epidemiological situation of a region. Moreover, such profiles allowed for convenient follow-up of infection after a focus as a function of time and geographical spread. PMID- 12721799 TI - The pseudorabies virus immediate-early promoter directs neuronal tissue-specific expression in transgenic mice. AB - Pseudorabies virus (PRV) immediate-early (IE) gene product is required for expression of the viral early and late genes as a transactivator. The IE gene is expressed as the first gene among the viral genes after the infection. To examine the activity of the IE promoter in vivo, we have generated transgenic mice expressing transgenes under the control of the IE promoter. To analyze the tissue specificity of the transgene expression, mRNA of the transgene was monitored in various tissues from the transgenic mice by reverse transcriptase (RT)-polymerase chain reaction (PCR) analysis. A strong transgene expression was observed in the neuronal tissues by the RT-PCR analysis. These neuronal tissues included cerebrum, cerebellum and trigeminal nerve. Although the PCR product was hardly detected in other tissues by the RT-PCR analysis, specific PCR bands were detected in multiple organs (skin, skeletal muscles, heart muscles, lung, liver, spleen, small intestine and kidney) by Southern blot analysis using the RT-PCR products. These results indicate that although the IE promoter acts as a pan specific promoter in vivo, it is capable of driving a high level of transgene expression in neuronal tissues. PMID- 12721798 TI - Molecular analysis of the capsid coding region of a virulent encephalomyocarditis virus isolate after serial cell passages and assessment of its virulence. AB - EMCV has been recognized as a worldwide-shared pathogen in many host species with natural differences of virulence among strains. Differences of virulence were also demonstrated experimentally after serial passages of particular EMCV strains in cell culture. In order to evaluate the genetic variability of the CCR of an highly virulent Belgian EMCV isolate after 210 passages in cell culture, this region has been molecularly characterised by RT-PCR and sequencing. The analyses enlightened the stability of the isolate as only 3 mutations appeared within the CCR and this was observed between passage 200 and 210. One of the mutations is adjacent to the amino acid described as the differentiation amino acid between diabetogenic and non-diabetogenic variants of EMCV. An experimental inoculation of pigs with the passage 210 of the virus enlightened its attenuation and the challenged pigs were protected against the wild-type virus. Results confirm the ability of EMCV to become attenuated after passages in cell culture. PMID- 12721800 TI - Genomic RNA constellation of recently emerging serotype G14 equine rotavirus strains in Japan that is highly homologous with prototype G3 and G14 strains previously identified in the United States of America. AB - Serotype G14 was once considered to be uncommon among equine rotaviruses. While it sporadically emerged in some parts of the world, serotype G14 became the dominant G serotype among rotaviruses detected in foals with diarrhea in Japan in the late 1990s. However, it is not known how such recently emerging G14 rotaviruses are related in their overall genomic RNA constellation to prototype G14 strain identified earlier in the United States of America or how they were generated and why they have dominated over G3 equine rotaviruses. Genogrouping by RNA-RNA hybridization revealed that recently emerging serotype G14 equine rotavirus strains had an overall genomic RNA constellation that was highly conserved not only with contemporary and earlier G3 strains in Japan but also with prototype G3 and G14 strains previously identified in the United States of America. Japanese G14 rotavirus strains are likely to have originated form a VP7 gene substitution reassortant that had been formed earlier in the United States of America on the background of the then dominant G3 equine rotavirus. PMID- 12721801 TI - Genomic characterisation of taro bacilliform virus. AB - Taro bacilliform virus (TaBV) has been classified as a putative badnavirus based on its non-enveloped, bacilliform virion morphology and transmission by mealybugs. The complete nucleotide sequence of a Papua New Guinea isolate of TaBV has now been determined and comprises 7458 bp. The genome contains four open reading frames (ORFs) on the plus-strand that potentially encode proteins of 17, 16, 214 and 13 kDa. The size and organisation of TaBV ORFs 1-3 is similar to that of most other badnaviruses, while the location of ORF 4 is similar to that of ORF 4 and ORF X of the atypical badnaviruses Citrus yellow mosaic virus and Cacao swollen shoot virus, respectively. The putative amino acid sequence of TaBV ORF 3 contained motifs that are conserved amongst badnavirus proteins including aspartic protease, reverse transcriptase (RT) and ribonuclease H (RNase H). The highly conserved putative plant tRNA(met)-binding site was also present in the 935 bp intergenic region of TaBV. Phylogenetic analysis using the amino acid sequence of ORF 3 showed that TaBV branched most closely to Dioscorea bacilliform virus. These results confirm that TaBV is a pararetrovirus of the genus Badnavirus, family Caulimoviridae. PMID- 12721802 TI - Adenoviral-expressed GP5 of porcine respiratory and reproductive syndrome virus differs in its cellular maturation from the authentic viral protein but maintains known biological functions. AB - The ORFs 5, 6 and 7, encoding for the three major structural proteins, GP(5), M and N, of the IAF-Klop strain of PRRSV were cloned and expressed in 293 cells using replication-defective human type 5 adenoviral vectors (hAdVs). Although the M protein gene could be cloned into hAdVs and expressed constituvely in 293 cells under the control of the hCMV immediate early promotor/enhancer, hAdVs expressing N and GP(5) proteins, which appeared to be toxic or interfered with adenovirus replication, could only be generated by inclusion of a tetracycline-regulatable promotor in the transfer vector pAdTR5. The recombinant (rec) proteins appeared similar to the authentic viral proteins in regards to their M(r)s and antigenicities. However, the recGP(5) apparently possesses different N-linked oligosaccharides residues. Its sensitivity to endo-beta-galactosidase digestion indicates that poly-N-acetyllactosamine is present on the individually-expressed protein, but not on the authentic GP(5) anchored into the virion envelope. The recGP(5) apparently accumulates within the ER compartment as a glycoprotein that possesses high-mannose N-linked oligosaccharide side chains sensitive to endo beta-N-acetylglucosaminidase H treatment, by contrast to its viral counterpart for which N-linked oligosaccharide side chains are of both high-mannose and complex types. Coinfection of 293 cells with hAdVs expressing the M and GP(5) did not lead to M-GP(5) heterodimer formation, as demonstrated in PRRSV-infected cells. Moreover, cells infected with inducible hAdV/ORF5 showed that GP(5) of the North American strain is proapoptotic. Indeed, when the expression cassette was turned-on, caspase 3 activity in hAdV/ORF5 infected cells was enhanced and DNA fragmentation could be detected by TUNEL assays. Pigs intradermally injected twice with hAdV/ORF5 developed antibody titers to the authentic viral GP(5) as soon as 10 days following challenge with the homologous virulent PRRSV strain, as revealed by Western blot and virus neutralization tests, suggesting the establishment of a specific immune memory. PMID- 12721803 TI - Infection of macaques with an R5-tropic SHIV bearing a chimeric envelope carrying subtype E V3 loop among subtype B framework. AB - To establish simian/human immunodeficiency virus (SHIV) clones bearing a chimeric envelope carrying subtype E V3 loop among subtype B envelope, four subtype E V3 sequences were substituted into SHIV(MD14), a SHIV clone bearing an envelope derived from a CXCR4 (X4)/CCR5 (R5)-dual tropic subtype B HIV-1 strain. SHIV TH09V3, an only V3-chimera clone capable of replicating in human and macaque peripheral blood mononuclear cells (PBMCs), was propagated in pig-tailed macaque PBMCs and in cynomolgus macaque splenic mononuclear cells. The propagated virus stocks were intravenously inoculated into respective macaque species. SHIV-TH09V3 infected both macaque species as shown by plasma RNA viremia, isolated viruses from PBMCs and plasma, and antibody production against viral proteins. To assess how the substituted V3 sequence affected coreceptor usage, SHIV-TH09V3 stocks propagated in vitro and after isolation from macaques were verified for their corecepor usage by GHOST cells assay. SHIV-TH09V3 maintained R5-tropic phenotype both in vitro and after isolation from macaques, in contrast to the X4/R5-dual tropic SHIV(MD14). This indicates the substituted V3 sequence among the backbone of SHIV(MD14) governs coreceptor usage. Future study of infecting macaques with SHIV-TH09V3 and SHIV(MD14) will focus on differences of the outcome caused by the different V3 sequences in connection with coreceptor usage. PMID- 12721804 TI - Sequence and genetic arrangement of the UL region of the monkey B virus (Cercopithecine herpesvirus 1) genome and comparison with the UL region of other primate herpesviruses. AB - The complete DNA sequence of the unique long (U(L)) region of monkey B virus (BV) was determined. Based on sequence homology and the presence of transcriptional control element motifs, homologues of every open reading frame present in the U(L) region of the Human herpesvirus 1 (herpes simplex virus 1, HSV-1) and Human herpesvirus 2 (herpes simplex virus 2, HSV-2) genomes were identified in BV. The BV genes are arranged in the same order and orientation as in HSV. These results demonstrate that the BV U(L) region is entirely co-linear with that of HSV-1 and HSV-2. PMID- 12721805 TI - Temperature sensitive mutations in the genes encoding the NS1, NS2A, NS3, and NS5 nonstructural proteins of dengue virus type 4 restrict replication in the brains of mice. AB - Using reverse genetics, it is possible to readily add well-defined attenuating mutations to the genome of wild type or incompletely attenuated dengue (DEN) viruses to generate vaccine candidates that exhibit the desired balance between attenuation and immunogenicity. Here, we describe the identification of eight temperature sensitive missense mutations distributed in four non-structural protein genes that specify a 60- to 10,000-fold range of restricted replication in the suckling mouse brain compared to wild type recombinant DEN4 virus. The usefulness of such mutations in flavivirus vaccine design and development is discussed. PMID- 12721806 TI - Characterization of a distinct carlavirus isolated from Verbena. AB - Avirus was isolated from Verbena plants that bore virus-like symptoms. The virus, for which the name Verbena latent virus (VeLV) is proposed, was consistently isolated from these plants, both with and without disease symptoms. Electron microscopy studies of ultrathin sections of infected Verbena tissues revealed the presence of elongated flexuous virus particles, ca. 650 nm in length. Its experimental host range was limited to Verbena spp. and Nicotiana clevelandii. No inclusion bodies or specific cytopathological effects, were observed. Electrophoresis of dissociated purified virus preparation in sodium dodecyl sulfate-polyacrylamide gel revealed a major protein component with a molecular mass of 38.9 kDa. Polyclonal antibodies which could specifically bind to virus particles were produced. A portion of the viral RNA was cloned and sequenced; it comprised 2503 nucleotides and contained part of three open reading frames (ORFs) which from the 5' to the 3'-ends, potentially encode for 489 amino acids (ORF1), a 25.8-kDa protein (ORF2) and a 12-kDa protein (ORF3). Comparison of the predicted amino acid sequence with those of other plant viruses revealed 40-60% identity with several carlaviruses. In the light of particle morphology, absence of specific cytopathological effects in ultrathin sections, and genomic and serological properties, it is suggested that this virus belongs to the genus Carlavirus. PMID- 12721807 TI - Activation of defense-related gene expression and systemic acquired resistance in cucumber mosaic virus-infected tobacco plants expressing the mammalian 2'5'oligoadenylate system. AB - Tobacco plants expressing the mammalian 2'5'oligoadenylate system (2-5A system) exhibit resistance to cucumber mosaic virus (CMV). Here, to characterize the molecular aspect of the resistance to CMV in 2-5A system-expressing tobaccos, the activation of defense-related genes and systemic acquired resistance (SAR) as the markers for the hypersensitive resistance (HR), were elucidated. Northern hybridization analysis indicated that the expression of four pathogenesis-related (PR) protein genes and five HR-related genes were induced in CMV-infected tobaccos expressing 2-5A system. Furthermore, the induction of SAR against Pseudomonas syringae pv. tabaci as second challenge, was observed on CMV inoculated tobaccos expressing 2-5A system. These results suggested that the resistance to CMV in tobacco expressing 2-5A system is associated with the establishment of an HR-like response. PMID- 12721808 TI - Regional variations in the physiology of the rat caudate-putamen. 1. In vivo subregional effects of amphetamine on extracellular dopamine concentration. AB - The caudate putamen is a neurochemically and functionally heterogeneous nucleus. Understanding the correlation between these regional variations in neurochemistry and function could greatly aid in the treatment of neurological disorders associated with this area of the brain. Since dopaminergic dysfunction has been implicated in some of these disorders, regional variations in the neurochemistry of this transmitter system are of particular interest. The dopaminergic response to 2.5 mg/Kg D-amphetamine was examined by in vivo voltammetry in 7 dorsal and 7 ventral regions of the caudate-putamen in the urethane anaesthetized rat. Extracellular dopamine concentration increased in all the areas examined. However, the effect was regionally heterogeneous-areas separated by as little as 1 mm showing significantly different responses in terms of both the absolute change and the rate of change in dopamine concentration. A significant general trend was also evident. Amphetamine produced an increasing effect in extracellular dopamine concentrations in the dorsal and lateral areas of the nucleus. It was concluded that the regionally heterogeneous effects of amphetamine on extracellular dopamine could be attributed to regional variations in the density of dopamine transport sites within the caudate-putamen. Since this transport site is the site of entry of a number of neurotoxins this finding may contribute to our understanding of the functional loss associated with disorders such as Parkinson's disease. PMID- 12721809 TI - Regional variations in the physiology of the rat caudate-putamen. 2. Effects of amphetamine and amphetamine induced dopamine release on basal and cortical stimulation evoked multiple unit activity. AB - Dopaminergic terminals within the caudate-putamen are located in an ideal position to modulate the corticostriatal system. Since this is the major afferent system of the striatum, dopamine has very powerful effects on striatal electrophysiological activity. The striatum is a regionally specialized multifunctional nucleus. It is therefore important to determine if dopamine has the same modulatory effects within different areas of the nucleus. The effects of 2.5 mg/Kg D-amphetamine (IP) on cortical stimulation evoked and basal multiple unit activity (MUA) was measured in 7 dorsal and 7 ventral striatal areas of the urethane anaesthetized rat. In general, amphetamine caused an increase in the basal activity and a decrease in the cortical stimulation evoked activity. However, there were both qualitative and quantitative regionally dependent differences in these responses. The effect on basal MUA was more pronounced in the dorsal and caudal areas whereas the effect on cortical stimulation evoked MUA was more pronounced in the ventral areas. The electrophysiological effects of amphetamine within the striatum were correlated with its regionally dependent effects on extracellular dopamine. This produced a measure of the effects of striatal dopamine on regional electrophysiological activity. This information was also used to determine the mathematical relationship between dopamine concentration change and the change in MUA. These data indicate that the excitatory effects of amphetamine-induced dopamine release on the non-stimulated MUA progressively increase along the rostro-caudal axis of the nucleus. In addition, the effects were more pronounced in the ventromedial as compared to the ventrolateral areas. These effects correlated best with the rate of change in dopamine concentration. In the dorsal striatum amphetamine-induced increases in dopamine had a regionally homogeneous inhibitory effect on the stimulated MUA. In the ventral striatum however, it had a progressively stronger effect along the rostro-caudal axis. These effects correlated best with the absolute change in dopamine concentration. PMID- 12721810 TI - Striatal formation of 6-hydroxydopamine in mice treated with pargyline, pyrogallol and methamphetamine. AB - Formation of 6-hydroxydopamine (6-OHDA) has been posited in the striatum following methamphetamine treatment and plays a critical role in methamphetamine induced nigrostriatal dopaminergic toxicity. We used high performance liquid chromatography electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) to determine the formation of 6-OHDA by the treatments of methamphetamine combined with pargyline, a monoamine oxidase inhibitor, and pyrogallol, a catechol-O-methyl-transferase inhibitor, in female C57BL/6J mouse striatum. A substantial amount of 6-OHDA (9.9 +/- 0.7 pg/mg wet tissue) was detected in mice treated with pargyline (100 mg/kg) and pyrogallol (25 mg/kg) in combination. Greater striatal 6-OHDA levels were observed in mice treated with combined pargyline, pyrogallol and methamphetamine (50 mg/kg) as compared to mice treated with combined pargyline and pyrogallol. However, mice treated with pargyline and pyragollol in combination followed by one and two doses of methamphetamine exhibited comparable striatal 6-OHDA levels (23.2 +/- 4.3, 27.3 +/- 1.3 pg/mg wet tissue) in our protocol. We conclude that blockade of the primary metabolic pathways of dopamine by inhibiting both monoamine oxidase and catechol-O-methyl transferase activities is sufficient to induce 6-OHDA formation in the striatum. Acute 6-OHDA accumulation in the striatum can be potentiated by methamphetamine, a potent dopamine releaser, administration following such metabolic inhibitions. PMID- 12721811 TI - BOLD fMRI response to direct stimulation (transcranial magnetic stimulation) of the motor cortex shows no decline with age. AB - Previous studies using BOLD fMRI to examine age-related changes in cortical activation used tasks that relied on peripheral systems to activate the brain. They were unable to distinguish between alterations due to age-related changes in the periphery and actual changes in cortical physiology. Transcranial magnetic stimulation (TMS), which allows direct, noninvasive stimulation of cortical neurons, was interleaved with BOLD fMRI to study 6 young and 5 old subjects. Three different tasks were compared: direct stimulation by TMS, indirect active stimulation produced by a motor task, and indirect passive stimulation produced by hearing the TMS coil discharge. Direct neuronal stimulation by TMS produced similar fMRI signal increases in both groups, suggesting that cortical physiology itself may not necessarily decline with age. PMID- 12721812 TI - Neuroprotection by deprenyl and other propargylamines: glyceraldehyde-3-phosphate dehydrogenase rather than monoamine oxidase B. AB - Deprenyl and other propargylamines are clinically beneficial in Parkinson's disease (PD). The benefits were thought to depend on monoamine oxidase B (MAO-B) inhibition. A large body of research has now shown that the propargylamines increase neuronal survival independently of MAO-B inhibition by interfering with apoptosis signaling pathways. The propargylamines bind to glyceraldehyde-3 phosphate dehydrogenase (GAPDH). The GAPDH binding is associated with decreased synthesis of pro-apoptotic proteins like BAX, c-JUN and GAPDH but increased synthesis of anti-apoptotic proteins like BCL-2, Cu-Zn superoxide dismutase and heat shock protein 70. Anti-apoptotic propargylamines that do not inhibit MAO-B are now in PD clinical trial. PMID- 12721813 TI - Idiopathic Parkinson's disease: possible routes by which vulnerable neuronal types may be subject to neuroinvasion by an unknown pathogen. AB - The progressive, neurodegenerative process underlying idiopathic Parkinson's disease is associated with the formation of proteinaceous inclusion bodies that involve a few susceptible neuronal types of the human nervous system. In the lower brain stem, the process begins in the dorsal motor nucleus of the vagus nerve and advances from there essentially upwards through susceptible regions of the medulla oblongata, pontine tegmentum, midbrain, and basal forebrain until it reaches the cerebral cortex. With time, multiple components of the autonomic, limbic, and motor systems become severely impaired. All of the vulnerable subcortical grays and cortical areas are closely interconnected. Incidental cases of idiopathic Parkinson's disease may show involvement of both the enteric nervous system and the dorsal motor nucleus of the vagus nerve. This observation, combined with the working hypothesis that the stereotypic topographic expansion pattern of the lesions may resemble that of a falling row of dominos, prompts the question whether the disorder might originate outside of the central nervous system, caused by a yet unidentified pathogen that is capable of passing the mucosal barrier of the gastrointestinal tract and, via postganglionic enteric neurons, entering the central nervous system along unmyelinated praeganglionic fibers generated from the visceromotor projection cells of the vagus nerve. By way of retrograde axonal and transneuronal transport, such a causative pathogen could reach selectively vulnerable subcortical nuclei and, unimpeded, gain access to the cerebral cortex. The here hypothesized mechanism offers one possible explanation for the sequential and apparently uninterrupted manner in which vulnerable brain regions, subcortical grays and cortical areas become involved in idiopathic Parkinson's disease. PMID- 12721814 TI - Preserved memory traces within diencephalic amnesia. AB - A male patient with bilateral thalamic lesions (medio-ventral nuclei) was investigated. Despite explicit memory impairments his lexical ability was normal. We recorded magnetic field changes (magnetoencephalography, MEG) during the performance of an animate/inanimate discrimination task in which some words where repeated after long delays. Normally, repeated items are classified significantly faster than their first presentations which is accomplished by an unconscious process called priming. The patient did not show any behavioural evidence of priming but the physiological data indicated preservation of this robust form of memory. Brain activation associated with repetitions was attenuated at early stages. The activity difference was posteriorly distributed which is consistent with previous reports about repetition priming. The present study indicated that the bilateral thalamic lesions of our patient disconnected the information processing stream between the primed information and the behavioural response. PMID- 12721815 TI - Olanzapine, risperidone and haloperidol in the treatment of adolescent patients with schizophrenia. AB - OBJECTIVES: To evaluate and compare the drug response and side effects of adolescents with schizophrenia treated with olanzapine, risperidone, and haloperidol. METHODS: Forty-three patients were treated with olanzapine (n = 19), risperidone (n = 17) and haloperidol (n = 7) for 8 weeks in an open clinical trial. Clinical improvement was evaluated with the Positive and Negative Syndrome Scale (PANSS), and side effects with the Udvalg for Kliniske Undersogelser (UKU) Side Effect Rating Scale. RESULTS: Significant clinical improvement was observed by week 4 for all medications. Olanzapine and haloperidol induced fatigability more frequently than risperidone. Haloperidol was associated with a higher frequency of depression and more severe extrapyramidal symptoms. CONCLUSIONS: To the best of our knowledge this is the first study in adolescents to compare the efficacy and side effects of three most commonly prescribed antipsychotic medications. Olanzapine, risperidone and haloperidol appear to be equally effective for the treatment of schizophrenia in adolescent inpatients but have different side effect profiles. PMID- 12721816 TI - Dopamine D3 receptor gene polymorphism and violent behavior: relation to impulsiveness and ADHD-related psychopathology. AB - Several lines of evidence indicate that dopaminergic neurotransmission is involved in the regulation of impulsive aggression and violence and that genetically determined variability in dopaminergic gene expression modifies complex traits including that of impulsivity and aggression. In this study we report an association of the dopamine D3 receptor (DRD3) polymorphism with impulsiveness according to Eysenck's EIQ and scores on the German short version of the Wender Utah Rating Scale (WURS-k), which we used for the assessment of a history of ADHD-related symptoms. This association was detected in a group of violent offenders, but not in non-violent individuals. Highest scores of EIQ impulsiveness and of the WURS-k were found in heterozygous violent individuals, while homozygotes showed significant lower rating scores, suggesting an heterosis effect. The results of our study suggest that variations of the DRD3 gene are likely involved in the regulation of impulsivity and some psychopathological aspects of ADHD related to violent behavior. PMID- 12721817 TI - Morphological characterization of the mycorrhiza formed by Helianthemum almeriense Pau with Terfezia claveryi Chatin and Picoa lefebvrei (Pat.) Maire. AB - This work presents the first anatomical description of the mycorrhizal systems of Helianthemum almeriense, and of the structure and ultrastructure of the mycorrhizae formed by this plant species with the ascomycetes Terfezia claveryi and Picoa lefebvrei. Four different mycorrhizal systems are described, the club shaped mycorrhiza being the most abundant. The type of mycorrhiza formed depended on the mycorrhiza culture conditions, but not on the fungal species. For both fungal species, H. almeriense formed an endomycorrhiza in natural field conditions, an ecto- and ectendomycorrhiza without a sheath in pot cultures, and an ectomycorrhiza with a characteristic sheath and Hartig net in in vitro cultures. This is the first report of a typical sheath in Helianthemum-desert truffle mycorrhizal associations. The results support the idea that culture conditions can induce changes in mycorrhiza morphology and that there is no clear barrier between the two main types of mycorrhiza organization in Helianthemum species. The ultrastructural study confirmed the regular presence of T. claveryi intracellular hyphae in direct contact with the host wall, a localization which seems to be a characteristic of the T. claveryi mycorrhiza organization. The P. lefebvrei mycorrhiza organization was characterized by intracellular hyphae with large amounts of electron-dense globules, probably with a lipidic content, and a warty ornamentation on the wall of the root external hyphae. PMID- 12721818 TI - Interactions between Trichoderma pseudokoningii strains and the arbuscular mycorrhizal fungi Glomus mosseae and Gigaspora rosea. AB - The interaction between Trichoderma pseudokoningii (Rifai) 511, 2212, 741A, 741B and 453 and the arbuscular mycorrhizal fungi Glomus mosseae (Nicol. & Gerd.) Gerdemann & Trappe BEG12 and Gigaspora rosea Nicolson & Schenck BEG9 were studied in vitro and in greenhouse experiments. All T. pseudokoningii strains inhibited the germination of G. mosseae and Gi. rosea except the strain 453, which did not affect the germination of Gi. rosea. Soluble exudates and volatile substances produced by all T. pseudokoningii strains inhibited the spore germination of G. mosseae. The germination of Gi. rosea spores was inhibited by the soluble exudates produced by T. pseudokoningii 2212 and 511, whereas T. pseudokoningii 714A and 714B inhibited the germination of Gi. rosea spores by the production of volatile substances. The strains of T. pseudokoningii did not affect dry matter and percentage of root length colonization of soybean inoculated with G. mosseae, except T. pseudokoningii 2212, which inhibited both parameters. However, all T. pseudokoningii strains decreased the shoot dry matter and the percentage of AM root length colonization of soybean inoculated with Gi. rosea. The saprotrophic fungi tested seem to affect AM colonization of root by effects on the presymbiotic phase of the AM fungi. No influence of AM fungi on the number of CFUs of T. pseudokoningii was found. The effect of saprotrophic fungi on AM fungal development and function varied with the strain of the saprotrophic species tested. PMID- 12721819 TI - Procalcitonin as a marker of acute pyelonephritis. PMID- 12721820 TI - UV-B radiation arising from stratospheric ozone depletion influences the pigmentation of the Antarctic moss Andreaea regularis. AB - Changes to the radiative environment arising from stratospheric ozone (O(3)) depletion and subsequent associations between these changes and the pigmentation of the moss Andreaea regularis were measured in late austral spring and early summer 1998 at Rothera Point on the western Antarctic Peninsula (67 degrees S, 68 degrees W). A strong relationship between O(3) column depth and the ratio of UV-B to PAR irradiance ( F(uv-b)/ F(par)) was recorded at ground level ( r(2)=92%, P<0.001). Weaker, but significant, associations between O(3) column depth and ground level unweighted and biologically effective UV-B radiation (UV-B(be)) were also found. Regression analyses indicated that F(uv-b)/ F(par) was the best predictor for concentrations of UV-B screening pigments and total carotenoids extracted from plant tissues. Concentrations of these pigments were loosely ( r(2)= ca. 30%) but significantly ( P<0.01) positively associated with F(uv-b)/ F(par). Concentrations of UV-B screening pigments were also positively associated with irradiances and daily doses of unweighted UV-B and UV-B(be) radiation. The concentrations of chlorophylls a and b were apparently unaffected by O(3) depletion. The data derived from this study suggest that changes to the radiative environment associated with stratospheric O(3) depletion influence the pigmentation of A. regularis. As a corollary, flavonoids are shown to be present in tissues of A. regularis. PMID- 12721822 TI - Growth reduction of Sphagnum magellanicum subjected to high nitrogen deposition: the role of amino acid nitrogen concentration. AB - We tested the relationship between Sphagnum growth and the amount of nitrogen stored in free amino acids in a fertilisation experiment with intact peat monoliths in an open greenhouse in The Netherlands. Three nitrogen deposition scenarios were used: no nitrogen deposition, field conditions and a doubling of the latter, corresponding to 0, 40 and 80 kg N ha(-1 )year(-1). Growth of Sphagnum as expressed by height increment was reduced in the 80 kg N treatment, but showed no correlation with the total nitrogen tissue concentration or with the concentration of individual or pooled free amino acids. The amount of nitrogen stored in free amino acids increased concomitantly with deposition, although it lagged more and more behind the total nitrogen concentration, the latter pointing to the accumulation of unmeasured nitrogen compounds. Asparagine clearly acted as the major storage compound for nitrogen in Sphagnum stem tissue, whereas arginine fulfilled this function to a lesser extent in the capitulum. It appears that nitrogen-induced growth inhibition of Sphagnum is related to acclimation rather than to certain threshold concentrations of amino nitrogen or total nitrogen. We propose that when Sphagnum is exposed to a step increase of nitrogen, its nitrogen metabolism does not adapt fast enough to keep up with the enhanced uptake rate. This imbalance between nitrogen uptake and assimilation may lead to an accumulation of toxic NH(4)(+ )in the cell and a subsequent reduction in growth. PMID- 12721821 TI - Food quality controls reproduction of the zebra mussel (Dreissena polymorpha). AB - Species such as Dreissena polymorpha sometimes contribute substantially in the transfer of primary to secondary production. During the ontogenetic cycle, the reproductive investment of adult mussels is one of the main parameters that affect recruitment success. We studied how food quality and temperature affect the reproductive investment in term of egg mass of D. polymorpha in a lake by sampling mussels monthly from 4 m and 15 m depths. Temperature affected reproduction directly and also indirectly through the food. To assess whether temperature and/or food conditions led to the differences observed in mussels sampled from the two depths, mussels were reared in the laboratory under two different temperature regimes for 3 months, simulating the temperature of the lake at 4 m and 15 m depth. Possible effects of food quality were tested at each temperature using four diets differing in fatty acid composition. Temperature played an important role as a trigger for spawning, and the type of diet clearly affected the reproductive investment. When the heterokont chromophyte alga Nannochloropsis limnetica, which is rich in polyunsaturated fatty acids (PUFAs) and long-chained PUFAs (>C18), was fed to mussels, an increased egg mass was obtained. This result was in contrast to that found when the green alga Scenedesmus obliquus and the cyanobacterium Aphanothece sp., both of which are deficient in long-chained PUFAs, were offered as food to the mussels. Such a PUFA dependent food quality may affect reproduction in lakes. Food quality effects vary seasonally in a lake and may be most important in summer, when low-food quality green algae and cyanobacteria are abundant. The low biochemical quality of these blooms may affect at least the later period of gametogenesis of D. polymorpha, which reproduces from June to August. PMID- 12721823 TI - Decelerating and sex-dependent tooth wear in Norwegian red deer. AB - In ungulates, tooth wear is often suggested as a proximate cause of senescence. Tooth wear is expected to be sex-dependent since energetic requirements and food selection varies largely between sexes in sexually dimorphic ungulates. Furthermore, tooth wear may lower mastication efficiency, and we predict a negative correlation between tooth wear and body weight or condition. We tested these predictions on data on tooth wear (estimated as height of first molar) of 1,311 male and 1,348 female red deer ( Cervus elaphus) aged 3-25 years and harvested along the west coast of Norway. The rate of tooth wear decreased with age. Males wear teeth at a higher rate (from 0.61 mm/year in 4-year olds to 0.45 mm/year in 11-year olds) than females (from 0.52 mm/year in 4-year olds to 0.39 mm/year in 11-year olds). Molar height correlated positively with body weight in both sexes, but not after adjusting for body size. Molar height was strongly dependent on body size in 3-year-old individuals (when tooth wear is minimal). Earlier reports in the literature of a positive correlation between tooth height and body weight may therefore be due to initial size differences rather than differences in condition due to tooth wear. PMID- 12721825 TI - Interactions between parasitized and unparasitized conspecifics: parasitoids modulate competitive dynamics. AB - Parasitism influences many aspects of a host's behavior and physiology. Therefore, parasitism is also likely to influence the competitive ability of the host. Field populations of phytophagous insects are often a mix of parasitized and unparasitized conspecifics and the inclusion of parasitism in their competitive dynamics may alter expected outcomes. We investigated the influence of parasitism by the hymenopteran parasitoid Phanerotoma franklini Gahan on the competitive interactions among larvae of its host Acrobasis vaccinii Riley. We found that parasitized larvae were poorer competitors and required less food to complete development compared to unparasitized larvae. To examine the influence of parasitism on the competitive dynamics of this system, we constructed an individual-based model parameterized with our laboratory data. The model examined the role of resource availability and parasitism rate on larval survival. The model suggests that parasitized larvae (and, hence parasitoids) experience higher levels of mortality from competition than unparasitized larvae. Further, the model also suggests that the decreased consumption of resources by parasitized larvae results in a decline in the occurrence of competition as the parasitism rate increases. We suggest that these observations may be general to many parasitoid-host systems. PMID- 12721824 TI - Performance of the cyclic autumnal moth, Epirrita autumnata, in relation to birch mast seeding. AB - The mast depression hypothesis has been put forward to explain the 9- to 10-year population cycle of the autumnal moth (Epirrita autumnata; Lepidoptera: Geometridae) in northern Fennoscandia. We analysed long-term data from Finnish Lapland in order to evaluate the critical assumption of the mast depression hypothesis: that better individual performance of herbivores, followed by high annual growth rate of populations, occurs in the year following mast seeding of the host, the mountain birch ( Betula pubescens ssp. czerepanovii). Since mast seeding has been suggested to occur at the expense of chemical defence against herbivores, we bioassayed the quality of birch leaves from the same trees by means of yearly growth trials with autumnal moth larvae. We also measured the size of wild adults as a determinant of potential fecundity of the species in different years. The relative growth rate of larvae was poorer in post-mast years compared to other years, rather than better as assumed by the hypothesis. Conversely, a slight indication of the increase in potential fecundity was observed due to the somewhat larger adult size in post-mast years. Population growth rate estimates, however, showed that the increase in fecundity would have to be much higher to facilitate population increase towards a cyclic peak with outbreak density. Accordingly, our two data sets do not support the assumption of a higher annual growth rate in autumnal moth populations subsequent to mast seeding of the host, thereby contradicting the predictions of the mast depression hypothesis. Temperatures, when indexed by the North Atlantic Oscillation and accumulated thermal sums, were observed to correlate with the abundance or rate of population change of the autumnal moth. The factors underlying the regular population cycles of the autumnal moth, however, remain unidentified. Overall, we suggest that the causal agents in cyclic insect population dynamics should be clarified by field experimentation, since trophic interactions are complex and are further modified by abiotic factors such as climate. PMID- 12721826 TI - Is reproduction of the Australian house mouse (Mus domesticus) constrained by food? A large-scale field experiment. AB - Food quantity and especially food quality are thought to be key factors driving reproductive changes in the house mouse, Mus domesticus, leading to outbreaks of house mouse populations in the Australian grain-growing region. Characteristic changes during an incipient mouse plague are an early start of breeding, a high proportion of females breeding at a young age and a prolonged breeding season. We conducted a large-scale food manipulation during an incipient mouse plague, which started with early breeding and relatively high spring numbers of mice. We measured background food availability in four farms throughout the study and conducted a food manipulation experiment from November to March in two of them. After harvest in December 100-200 kg/ha spilled grain remained in the stubble. This was depleted by March. In two treatment farms we added high-protein food pellets on a weekly basis between November and March and two farms served as controls. We measured changes in mouse numbers by capture-mark-recapture trappings and changes in reproduction by scoring embryos and recent placental scars at necropsy. Mouse numbers did not differ between treatments and controls. There were no differences in the litter size or the proportion of females breeding between treatments and controls. We observed the normal pattern of high litter size in spring and decreasing litter size towards the end of summer in treatments and controls. In all farms reproduction stopped in March. Mouse numbers were high but not at plague densities. Contrary to our prediction we did not observe food constraint affecting the reproduction of female mice. Our field experiment seems to rule out food quality as the driving factor for improved reproduction and formation of an outbreak of mice. We suggest that physiological mechanisms in mice might not enable them to take advantage of food with a high protein content in arid summers in southeastern Australian grain fields because of the lack of free-standing water. PMID- 12721827 TI - Fruit abortion, developmental selection and developmental stability in Quercus ilex. AB - Fruit abortion has been hypothesized to be a parental means of selective removal of propagules with low viability. In particular, aborted zygotes have been suggested to have developmentally deviant phenotypes, and surviving offspring may therefore give rise to adults with a developmentally stable phenotype. We tested predictions from this hypothesis using acorns of holm oaks Quercus ilex as a model system. Fecundity of oak trees was negatively related to mean fluctuating asymmetry of leaves, and abortion rates were positively related to leaf fluctuating asymmetry in at least one population. Aborted acorns were asymmetric in 83-99% of cases in three samples, while mature acorns were only asymmetric in 57-78% of cases. Acorn asymmetry was unrelated to germination probability and germination date, and had no significant effect on number of leaves, leaf mass, stem mass, seedling height or leaf area of seedlings. However, acorn asymmetry affected the trade-off between number and size of leaves in seedlings. Seedlings from asymmetric acorns showed a positive relationship between acorn size and number of leaves, but no relationships between acorn size and leaf area, while symmetric acorns showed the opposite. A positive relationship between acorn size and number of leaves in spring was found for naturally emerged seedlings that died during their first summer, whereas the number of leaves produced by surviving seedlings did not depend on acorn size. These findings are consistent with the hypothesis that stressed trees selectively abort propagules of low viability, and that developmental selection acts on a measure of developmental instability of fruits. PMID- 12721828 TI - Interaction between maternal effects: onset of incubation and offspring sex in two populations of a passerine bird. AB - Maternal phenotype and maternal environment can profoundly affect the phenotype and fitness of offspring. Yet the causes of variation in such maternal effects are rarely known. Embryos in avian eggs cannot develop without being incubated and this creates an opportunity for maternal control of duration and onset of offspring development. However, females might adjust the start of incubation (e.g., coincident with the first egg or delayed until after egg-laying) in response to environmental conditions that they experience at the time of breeding. We studied two populations of the house finch ( Carpodacus mexicanus) that breed at the climatic extremes of the species' geographical range (Montana and Alabama) and found that in both populations, the timing of incubation onset was closely associated with the bias in the sequence in which male and female eggs were laid within a clutch. When females started incubation with the first egg, they produced sons and daughters in highly biased sequence, when females delayed the onset of incubation until after the egg-laying, the sequence of sons and daughters was not biased. Because in both populations, onset of incubation was associated with the ambient temperature, these results emphasize that maternal effects on offspring can be influenced by ecological conditions experienced by parental generation. PMID- 12721829 TI - Advantages of a mixed diet: feeding on several foliar age classes increases the performance of a specialist insect herbivore. AB - Two field studies were carried out to determine the influence of Abies balsamea foliage age on the preference and performance of larvae of Neodiprion abietis, a specialist Diprionid sawfly. Preference was determined by examining N. abietis defoliation on all age classes of foliage. Performance was estimated using larval survival, cocoon weights and the percentage of adults that were females. Neodiprion abietis preference for, and performance on, current-year foliage was very low, peaked on 2 or 3-year-old foliage, and declined on older foliage. Thus, sawfly feeding preference was adaptive. However, survival and cocoon weight were highest when sawflies were allowed to feed on all age classes of foliage, demonstrating that an insect specialist may perform better when feeding on several age classes of foliage from a single host plant species. These results indicate that either different larval instars have different nutritional requirements, or that food mixing provides the best diet, permitting the herbivore to obtain needed nutrients while avoiding ingestion of toxic doses of secondary metabolites. In addition, our results suggest that limited availability of varied foliage has more negative consequences for N. abietis females than for males, as the percentage of survivors that were females decreased when juvenile mortality was high. Our results emphasize the importance of considering non linear changes in foliar quality as leaves age on herbivore preference and performance, and demonstrate how a herbivore can use this variability to maximize its fitness. PMID- 12721830 TI - Effects of habitat disruption on the activity of nectarivorous bats (Chiroptera: Phyllostomidae) in a dry tropical forest: implications for the reproductive success of the neotropical tree Ceiba grandiflora. AB - In the tropical dry forest of the central Pacific coast of Mexico the pollination and reproductive success of the bombacaceous tree Ceiba grandiflora was negatively affected by habitat disruption. Two of the three bat species that function as effective pollinators for this species ( Glossophaga soricina and Musonycteris harrisoni) visited flowers found in trees in disturbed habitats significantly less than trees found in undisturbed habitats. A similar pattern was observed for the effective bat pollinator, Leptonycteris curasoae; however the difference was not significant. The three nectarivorous bats that functioned as effective pollinators of C. grandiflora also visited flowers to exclusively feed on pollen by biting or pulling off an anther (see Fig. S1 of Electronic Supplementary Material). The number of pollen grains deposited on stigmas from flowers in undisturbed areas was significantly greater than from flowers in disturbed habitats. The greater visitation rate and the greater number of pollen grains deposited on flowers from trees in undisturbed forest resulted in a significantly greater fruit set for trees in these areas. Our study demonstrates the negative effect that habitat disruption has on bat pollinators in tropical dry forest ecosystems and documents the negative consequences for the plants they pollinate. PMID- 12721831 TI - Bottom-up control of carabid beetle communities in early successional wetlands: mediated by vegetation structure or plant diversity? AB - Two hypotheses of bottom-up control that predict that the species richness of Carabidae will depend either on the taxonomic diversity of plants ("taxonomic diversity hypothesis") or on the structural heterogeneity of the vegetation ("structural heterogeneity hypothesis") were tested. Plant species were classified into nine plant structural groups through cluster analysis of morphological traits (e.g. total height) at 30 early successional temporary wetlands in the east-German agricultural landscape. In a linear regression analysis, the heterogeneity of vegetation structures explained 55% of the variation in carabid beetle diversity. According to a partial correlation analysis, plant taxonomic diversity did not have a significant effect, consistent with the "structural heterogeneity hypothesis," and contradicting previous studies which concluded that plant taxonomic diversity would be the most important factor in early successional habitats. An experimental study was used to test hypotheses on the processes underlying this bottom-up control by vegetation structure: the "hunting efficiency hypothesis," the "enemy-free space hypothesis," and the "microhabitat specialization hypothesis." The composition of plant structural groups in 15 vegetation plots (1 m(2)) was manipulated, creating a gradient from dense vegetation to open plots. Subsequent pitfall catches revealed significant differences in the activity-abundances of the carabid species. Large species preferred dense vegetation plots, consistent with the enemy-free space hypothesis that large species are more vulnerable to predation on the open plots and prefer dense vegetation to escape from natural enemies. The results indicate that bottom-up control is not mediated only by plant taxonomic or functional group diversity and that vegetation structures may be more important than previously suggested. PMID- 12721832 TI - Response of NDVI, biomass, and ecosystem gas exchange to long-term warming and fertilization in wet sedge tundra. AB - This study explores the relationship between the normalized difference vegetation index (NDVI), aboveground plant biomass, and ecosystem C fluxes including gross ecosystem production (GEP), ecosystem respiration (ER) and net ecosystem production. We measured NDVI across long-term experimental treatments in wet sedge tundra at the Toolik Lake LTER site, in northern Alaska. Over 13 years, N and P were applied in factorial experiments (N, P and N + P), air temperature was increased using greenhouses with and without N + P fertilizer, and light intensity (photosynthetically active photon flux density) was reduced by 50% using shade cloth. Within each treatment plot, NDVI, aboveground biomass and whole-system CO(2) flux measurements were made at the same sampling points during the peak-growing season of 2001. We found that across all treatments, NDVI is correlated with aboveground biomass ( r(2)=0.84), GEP ( r(2)=0.75) and ER ( r(2)=0.71), providing a basis for linking remotely sensed NDVI to aboveground biomass and ecosystem carbon flux. PMID- 12721833 TI - A comparative analysis of morphological and ecological characters of European aphids and lycaenids in relation to ant attendance. AB - Ants are a major environmental factor for many insect species. For example, aphids and lycaenids have evolved an array of associations with ants ranging from obligate myrmecophily to the avoidance of contact. Here we (1) analyze the predictive power of different ecological and morphological traits for explaining the strength of the association between ants and aphids/lycaenids and (2) contrast different taxonomic levels with respect to the variance explained by ant attendance. Data come from a literature survey including 112 species of aphids and 103 species of lycaenids from Europe. For aphids, feeding on woody plant parts is positively associated with ant attendance, while a high degree of mobility, feeding in isolation, and the possession of wings in the adult stage are negatively associated with ant attendance. In lycaenids, feeding on inflorescences and feeding on Fabaceae host plants is closely associated with ant attendance, while living in forests bears a smaller likelihood to establish mutualistic relationships. Body size always appeared to be a poor predictor for the degree of ant attendance. Overall, in both insect groups less than 10% of the variation in the ecological traits recorded is explained by the different modes of ant association. When decomposing the variance in traits explained by ant attendance at different taxonomic levels, aphids and lycaenids show contrasting results. In aphids, most variance in the degree of ant attendance is explained at the subfamily level and least at the species level. The opposite is true for lycaenids, where most variance is explained at the lowest taxonomic level. Possible mechanisms explaining these different patterns of associations with ants are suggested. PMID- 12721834 TI - Changes in habitat complexity negatively affect diverse gastropod assemblages in coralline algal turf. AB - The physical structure of a habitat generally has a strong influence on the diversity and abundance of associated organisms. I investigated the role of coralline algal turf structure in determining spatial variation of gastropod assemblages at different tidal heights of a rocky shore near Sydney, Australia. The structural characteristics of algal turf tested were frond density (or structural complexity) and frond length (the vertical scale over which structural complexity was measured). This definition of structural complexity assumes that complexity of the habitat increases with increasing frond density. While frond length was unrelated to gastropod community structure, I found significant correlations between density of fronds and multivariate and univariate measures of gastropod assemblages, indicating the importance of structural complexity. In contrast to previous studies, here there were negative relationships between the density of fronds and the richness and abundance of gastropods. Artificial habitat mimics were used to manipulate the density of fronds to test the hypothesis that increasing algal structural complexity decreases the richness and abundance of gastropods. As predicted, there were significantly more species of gastropods in loosely packed than in tightly packed turf at both low- and mid shore levels. Despite large differences between gastropod assemblages at different tidal heights, the direction and magnitude of these negative effects were similar at low- and mid-shore levels and, therefore, relatively independent of local environmental conditions. These novel results extend our previous understanding of the ecological effects of habitat structure because they demonstrate possible limitations of commonly used definitions of structural complexity, as well as distinct upper thresholds in the relationship between structural complexity and faunal species richness. PMID- 12721835 TI - Herbivorous mites as ecological engineers: indirect effects on arthropods inhabiting papaya foliage. AB - We examined the potential of a leaf roller to indirectly influence a community of arthropods. Two mite species are the key herbivores on papaya leaves in Hawaii: a spider mite, Tetranychus cinnabarinus Boisduval, and an eriophyid mite, Calacarus flagelliseta, which induces upward curling of the leaf margin at the end of the summer when populations reach high densities. A survey and three manipulative field experiments demonstrated that (1) leaf rolls induce a consistent shift in the spatial distribution of spider mites and their predators, the coccinellid Stethorus siphonulus Kapur, the predatory mites Phytoseiulus spp., and the tangle web building spider Nesticodes rufipes Lucas; (2) the overall abundance of spiders increases on leaves with rolls; (3) the specialist predators Stethorus and Phytoseiulus inhabit the rolls in response to their spider mite prey; and (4) the spider inhabits the rolls in response to the architecture of the roll itself. This study shows the importance of indirect effects in structuring a terrestrial community of herbivores. PMID- 12721836 TI - Reduced resistance of invasive varieties of the alien tree Sapium sebiferum to a generalist herbivore. AB - Invasive plants are often larger in their introduced range compared to their native range. This may reflect an evolved reduction in defense and increase in growth in response to low herbivory in their introduced range. Key elements of this scenario include genetic differences in defense and growth yet uniformly low rates of herbivory in the field that dissociate defense and herbivore damage for alien species. We conducted a laboratory experiment with Melanoplus angustipennis grasshoppers and Chinese Tallow Tree seedlings ( Sapium sebiferum) from its native range (China) and its introduced range (Texas, USA) where it is invasive. We caged grasshoppers with pairs of Sapium seedlings from the same continent or different continents. The amounts of leaf area removed from Texas and China seedlings, and their height growth rates, were indistinguishable when both seedlings in the pair were from the same continent. However, when grasshoppers had a choice between seedlings from different continents, they removed more Texas Sapium foliage than China Sapium foliage and height growth rates were higher for China Sapium seedlings compared to Texas seedlings. Grasshopper growth rates increased with greater Sapium foliage consumption. In a common garden in Texas, Sapium seedlings from Texas grew 40% faster than those from China. Chewing insect herbivores removed little Sapium foliage in the field experiment. Although grasshoppers preferred to feed on Texas Sapium when offered a choice in the laboratory, extremely low herbivory levels in the field may have allowed the Texas seedlings to outperform the China seedlings in the common garden. These results demonstrate post-invasion genetic differences in herbivore resistance and growth of an invasive plant species together with a decoupling of defense and herbivore choice in the introduced range. PMID- 12721837 TI - Avian community composition of kopjes in a heterogeneous landscape. AB - We examined avian communities of kopjes (naturally occurring insular rock outcrops) in grassland and thorn tree woodland habitats in the Seronera region of Serengeti National Park, northwestern Tanzania. Although kopjes cover a small proportion of the Park's area, they provide resources that are uncommon in the Serengeti landscape and are known to host diverse, yet poorly documented, biotic communities. The primary objectives of this study were (1) to compare avian communities of kopjes with those of their surrounding habitat matrixes; (2) to compare the avian communities among kopjes; and (3) to determine the effects of kopje characteristics (e.g., size, vegetation cover, level of human disturbance and matrix type) on avian diversity and community composition. The avian communities of kopjes differed significantly from those of the matrixes in species composition and guild characteristics. Rare species, frugivorous species and nectarivorous species were more abundant on kopjes, whereas there were more ground-feeding species in matrix sites. Species richness was positively correlated with the area of kopjes covered by tall vegetation (5+ m), but neither total habitat area nor total vegetation cover significantly affected avian diversity. The surrounding habitat type, the fruiting phenology of Ficus and Commiphora trees and the level of human disturbance also influenced the presence and abundance of individual species and accounted for differences in community composition among kopjes. Our results show that kopjes support unique avian assemblages comprising many species that are otherwise rare in Serengeti, and consequently that kopjes may be local hotspots of avian diversity in the region. PMID- 12721838 TI - Exposure of leopard frogs to a pesticide mixture affects life history characteristics of the lungworm Rhabdias ranae. AB - We tested the hypothesis that exposure of leopard frogs ( Rana pipiens) to agricultural pesticides can affect the infection dynamics of a common parasite of ranid frogs, the lungworm Rhabdias ranae. After a 21-day exposure to sublethal concentrations of a pesticide mixture composed of atrazine, metribuzin, aldicarb, endosulfan, lindane and dieldrin, or to control solutions (water, dimethyl sulfoxide), parasite-free juvenile frogs were challenged with 30 infective larvae of R. ranae. Approximately 75% of the larvae penetrated the skin and survived in both exposed and control animals, suggesting that pesticides did not influence host recognition or penetration components of the transmission process. Rather, we found that the migration of R. ranae was significantly accelerated in hosts exposed to the highest concentrations of pesticides, leading to the establishment of twice as many adult worms in the lungs of frogs 21 days post-infection. Pesticide treatment did not influence the growth of lungworms but our results indicate that they matured and reproduced earlier in pesticide-exposed frogs compared to control animals. Such alterations in life history characteristics that enhance parasite transmission may lead to an increase in virulence. Supporting evidence shows that certain components of the frog immune response were significantly suppressed after exposure to the pesticide mixture. This suggests that the immune system of anurans exerts a control over lungworm migration and maturation and that agricultural contaminants can interfere with these control mechanisms. Our results also contribute to the ongoing debate regarding the role that anthropogenic factors could play in the perplexing disease-related die-offs of amphibians observed in several parts of the world. PMID- 12721839 TI - Covariance of preference and performance on normal and novel hosts in a locally monophagous and locally polyphagous butterfly population. AB - Covariance between preference and performance was quantified for Papilio glaucus strains derived from a locally monophagous Florida population and a locally polyphagous Ohio population. I used two-choice assays to assess relative host preferences of mothers for plant species that represent reciprocal normal and novel hosts for each population (i.e., Liriodendron tulipifera and Magnolia virginiana) and a split-brood design to quantify relative performance of their progeny on each host. Covariance between preference and proxies of performance was detected independently within each population, which is a level of genetic structure at which such covariance has rarely been documented. These results support the hypothesis that preference-performance covariance can exist in populations that have no obvious internal host-associated structure. In the Ohio strain, all proxies of performance (larval duration, pupal mass, relative growth rate, and survival) were significantly correlated with relative preference for the normal host, L. tulipifera. In the Florida strain, however, only pupal mass was correlated with maternal preference, and this correlation was not in the direction expected. Progeny that attained the heaviest mass were derived from mothers that preferred L. tulipifera, the locally rare host. The nature of the preference-performance links was not in the manner predicted by conventional optimal oviposition theory, whereby host-associated tradeoffs have been considered an implicit element. Relative performance was congruent across hosts, regardless of whether mothers preferred L. tulipifera, M. virginiana, or neither host. After considering possible genetic and nongenetic explanations that could account for preference-performance covariance in P. glaucus, I conclude that this covariance has a genetic basis. Likely, multiple genetic control mechanisms (e.g., pleiotropy and co-adaptation) integrate at the level of different trait combinations and/or a particular trait combination to generate observed patterns. PMID- 12721840 TI - Mucopolysaccharidosis IVA: characterization of a common mutation found in Finnish patients with attenuated phenotype. AB - Mucopolysaccharidosis IVA (MPS IVA) is caused by the deficiency of the lysosomal enzyme N-acetylgalactosamine-6-sulfate sulfatase encoded by the GALNS gene on chromosome 16. We describe, in detail, the clinical phenotype of five patients from three unrelated Finnish families and have characterized the disease-causing mutations in GALNS. Genotypes of the patients are D60N/A291T, D60N/W230X, and D60N/1374delT. Mutation 1374delT introduces premature termination of GALNS. Cells over-expressing the novel mutation W230X and A291T had no residual GALNS activity, whereas D60N gave 12.2% residual activity compared with the wild type. Co-transfection of D60N/A291T and D60N/W230X showed 5.5% and 6.7% of wild type activity, respectively. The precursor proteins of D60N and A291T were observed at 55 kDa and 57 kDa, respectively, whereas there was no detectable band in cells over-expressing W230X. At 55 degrees C, the mutant protein showed lower thermostability than the wild type protein at pH 3.8 and 7.0. The tertiary structural model of the GALNS protein revealed that aspartic acid at position 60 is located on the surface of the molecule, away from the active site. This makes it unlikely that the enzymatic function of the protein with D60N is severely impaired. On the other hand, A291 and W230 are localized near the active site. The molecular characteristics of the D60N mutation explain the attenuated clinical phenotype of the patients. PMID- 12721841 TI - A point mutation in bioactive RNA results in the failure of mutant heart correction in Mexican axolotls. AB - Ambystoma mexicanum is an intriguing animal model for studying heart development because it carries a mutation in gene c. Hearts of homozygous recessive (c/c) mutant embryos do not contain organized myofibrils and fail to beat. The defect can be corrected by organ-culturing the mutant heart in the presence of RNA from anterior endoderm or endoderm/mesoderm-conditioned medium. By screening a cDNA library made of total conditioned medium RNA from normal axolotl embryonic endoderm, we isolated a single clone (MIR), the synthetic RNA from which corrects the mutant heart defect by promoting myofibrillogenesis and thus was named MIR (myofibrillogenesis inducing RNA). In the present study, we have examined MIR gene expression in mutant axolotl hearts at early pre-heart-beat developmental stages and found its quantitative expression, as detected by RT-PCR, to be the same as in normal hearts. However, careful analysis of sequence data revealed a G ->U point mutation in the mutant MIR RNA. Further computational analyses, using GENEBEE software to compare normal and mutant MIR RNAs show a significant alteration in RNA secondary structure of the point-mutated MIR RNA. The results from bioassay and confocal microscopy immunofluorescent studies demonstrate that, unlike MIR RNA derived from normal embryos, the mutated MIR RNA does not promote myofibrillogenesis in mutant embryonic hearts and fails to rescue/correct the mutant heart defect. PMID- 12721842 TI - Histomorphology of angiogenesis in human perinatal orbitofrontal cortex: a Golgi and electron microscopic study of anastomosis formation. AB - We demonstrate here for the first time the formation of vascular anastomoses in the human neocortex. The orbitofrontal cortex (OFC) of the perinatal human brain is structurally immature, so that the maturation of neurons, neuroglia and intracortical capillaries can be conveniently studied by comparison with these elements in motor and striatal cortex of the same brain. In OFC of the perinatal human brain, indication of prominent anastomosis formation in the upper layers (lamina II and III) is observed. Golgi silver impregnation and electron microscopic techniques were employed to demonstrate individual stages of vascular anastomosis development. Along with capillary angiogenesis, vascular anastomosis formation is of major importance for the ontogenesis of neuronal and glial elements of the CNS, particularly with respect to optimal metabolic support of structural elements of the mature brain. In the neocortex of the adult human brain, a characteristic three-dimensional capillary angioarchitecture is observed, contrasting with the two-dimensional arrangement in the immature perinatal cortex. Abnormal OFC angioarchitecture was also found in 14% of histomorphologically investigated brains from persons with endogenous psychoses. For the pathogenesis of functional disorders of the human brain, including endogenous psychoses, knowledge about the processes of structural maturation of neurons and neuroglia, but also of capillary architecture, is essential. PMID- 12721843 TI - Arabidopsis sulfurtransferases: investigation of their function during senescence and in cyanide detoxification. AB - Sulfurtransferases (STs) and beta-cyano- l-alanine synthase (CAS) are suggested to be involved in cyanide detoxification. Therefore, the accumulation of ST1 and CAS RNAs, and the ST and CAS protein levels and enzyme activities were determined in Arabidopsis thaliana Heynh. plants grown under different conditions. Senescence-associated processes were successfully induced by natural aging, by jasmonate methyl ester and by darkness in whole plants and detached leaves, as demonstrated by the expression of the senescence marker genes SAG12 and SAG13. However, the changes in RNA accumulation and protein levels of ST and CAS did not correlate with the expression of these senescence marker genes; the specific ST and CAS activities either decreased (ST) or increased (CAS). In another experiment, Arabidopsis plants were sprayed with cyanide to investigate the role of ST and CAS in cyanide detoxification. The expression of ST and CAS at the RNA and protein levels, and also the enzyme activities, remained equal in cyanide treated and control plants. Incubation with 1-aminocyclopropane-1-carboxylic acid, the precursor of ethylene, increased while fumigation with ethylene decreased expression and activity of ST and CAS. In summary, cyanide does not induce the expression or enhance the activity of ST and CAS in Arabidopsis. For both proteins the evidence for a role in cyanide detoxification or induced senescence is low. PMID- 12721844 TI - Effects of carbohydrate starvation on gene expression in citrus root. AB - The roots of alternate-bearing citrus (Murcott, a Citrus reticulata hybrid) trees undergo extreme fluctuations of carbohydrate abundance and starvation. Using this system, we investigated the effect of root carbohydrate (total soluble sugar, sucrose and starch) depletion on carbohydrate-related gene expression. A series of genes, including those coding for starch phosphorylase ( STPH-L and STPH-H), ADP-glucose pyrophosphorylase, small subunit ( Agps), R1, plastidic ADP/ATP transporter ( AATP), phosphoglucomutase ( PGM-P and PGM-C), sucrose synthase ( CitSuS1 and CitSuSA), sucrose transporter ( SUT1 and SUT2), hexokinase ( HK) and alpha-amylase ( alpha-AMY), have been isolated and their expression analyzed. The genes were found to respond differentially to carbohydrate depletion. STPH-L, STPH-H, Agps, R1, AATP, PGM-P, PGM-C, CitSuS1 and HK were down-regulated while SUT1 and alpha-AMY were up-regulated during carbohydrate depletion. Two other genes, CitSuSA and SUT2, did not respond to carbohydrate depletion. Fruit removal, which interrupted the carbohydrate depletion induced by heavy fruiting, reversed these gene expression patterns. Trunk girdling and whole-plant darkening treatments, which brought about root carbohydrate depletion, induced the same changes in gene expression obtained in the alternate-bearing system. The possible roles of the up- and down-regulated genes in the metabolism of carbohydrate depleted citrus roots are discussed. Although the specific signals involved have not been determined, the results support the feast/famine hypothesis of carbohydrate regulation proposed by Koch [K.E. Koch (1996) Annu Rev Plant Physiol Plant Mol Biol 47:509-540]. PMID- 12721846 TI - Mechanical, chemical and X-ray analysis of wood in the two tropical lianas Bauhinia guianensis and Condylocarpon guianense: variations during ontogeny. AB - Mechanical and chemical properties as well as microfibril angles of wood tissues from different ontogenetic stages are determined for the neotropical lianas Bauhinia guianensis and Condylocarpon guianense. The mechanical properties include the elastic moduli under bending and under dynamic torsion. The chemical analyses cover (i) the content of cellulose, lignin and hemicelluloses fractions, (ii) the monomeric composition of the uncondensed lignin, and (iii) the composition of the hemicelluloses with respect to neutral monosaccharides. By comparing the wood properties of these lianas with the corresponding properties of wood from self-supporting deciduous trees, common characteristics and differences are revealed. Additionally, the changes in the lignin and polysaccharides fractions as well as the variations in microfibril orientation that occur during ontogeny of the two liana species are discussed with regard to their implications for the mechanical properties of wood. PMID- 12721845 TI - Sm and U2B" proteins redistribute to different nuclear domains in dormant and proliferating onion cells. AB - Monoclonal antibodies against the spliceosomal proteins Sm and U2B", and against p105, a protein component of interchromatin granules, were used to investigate the nuclear distribution of the splicing factors in Allium cepa L. meristematic cells. Confocal microscopy showed that in steady-state proliferating cells, the spliceosomal components were distributed into two nuclear domains: (i) a diffuse nucleoplasmic network similar to that formed by interchromatin granules and (ii) numerous Cajal bodies. These domains were the counterpart of the perichromatin fibrils and granules, interchromatin granules and Cajal bodies observed by electron microscopy after EDTA and bismuth oxynitrate stainings. Dormant cells showed a nuclear distribution of the proteins in small Cajal bodies and numerous micro-speckles, correlated with the distribution of ribonucleoproteins (RNPs) observed by electron microscopy. The spliceosomal proteins relocated to the diffuse nucleoplasmic network and Cajal bodies when the cells were released from dormancy by water soaking and they re-started their proliferative activity. Inhibition of RNA synthesis by 5,6-dichloro-1-beta- d-ribofuranosylbenzimidazole (DRB) treatment in proliferating cells demonstrated that the micro-speckles were not the morphological expression of a transcription block. Fractionation and confocal microscopy studies showed a differential association of the splicing factors with the nuclear matrix depending not only on the protein, but also on nuclear activity. Our results suggest a reversible relocation of the spliceosomal proteins between different sub-nuclear domains in physiological conditions. We report here an unusual nuclear domain in dormant nuclei, the micro-speckles, corresponding to storage sites for RNPs, which were rapidly mobilised after water imbibition. PMID- 12721847 TI - Carbon and nitrogen distribution in the green algal lichens Hypogymnia physodes and Platismatia glauca in relation to nutrient supply. AB - With the aim of understanding how some lichens can survive intensive fertilization we investigated two green algal ( Trebouxia) lichens, Hypogymnia physodes (L.) Nyl. and Platismatia glauca (L.) W. Culb., and compared control (Ctr), and intensively fertilized (F) thalli. We measured total N, proteins and amino acids to assess lichen N status. Chlorophyll a indicated photosynthetic capacity and photobiont mass, ergosterol the metabolic demands of the fungus, and chitin the fungal biomass. For carbon status we measured glucose, the photobiont ( Trebouxia) export product ribitol, and the mycobiont-specific carbohydrates arabitol and mannitol. The F-thalli had 2-3 times higher protein and N concentrations, 5-10 times higher chlorophyll a concentrations, while ergosterol and chitin were doubled. The ribitol concentrations were 4-5 times higher in the F-thalli, while the fungal carbohydrates did not increase to the same extent. The amino acid arginine had increased 60-fold. The F-thalli also had a relatively higher N investment in the photobiont in relation to mycobiont tissue compared to the Ctr-thalli, probably resulting in an increased capacity for carbon assimilation, most possibly required for maintaining the higher nutrient status of the F-thalli. Arginine accumulation possibly avoided toxic effects of accumulated NH4+, albeit binding a significant fraction of assimilated carbon. PMID- 12721848 TI - tla1, a DNA insertional transformant of the green alga Chlamydomonas reinhardtii with a truncated light-harvesting chlorophyll antenna size. AB - DNA insertional mutagenesis and screening of the green alga Chlamydomonas reinhardtii was employed to isolate tla1, a stable transformant having a truncated light-harvesting chlorophyll antenna size. Molecular analysis showed a single plasmid insertion into an open reading frame of the nuclear genome corresponding to a novel gene ( Tla1) that encodes a protein of 213 amino acids. Genetic analysis showed co-segregation of plasmid and tla1 phenotype. Biochemical analyses showed the tla1 mutant to be chlorophyll deficient, with a functional chlorophyll antenna size of photosystem I and photosystem II being about 50% and 65% of that of the wild type, respectively. It contained a correspondingly lower amount of light-harvesting proteins than the wild type and had lower steady-state levels of Lhcb mRNA. The tla1 strain required a higher light intensity for the saturation of photosynthesis and showed greater solar conversion efficiencies and a higher photosynthetic productivity than the wild type under mass culture conditions. Results are discussed in terms of the tla1 mutation, its phenotype, and the role played by the Tla1 gene in the regulation of the photosynthetic chlorophyll antenna size in C. reinhardtii. PMID- 12721849 TI - Sweet pepper ferredoxin-like protein ( pflp) gene as a novel selection marker for orchid transformation. AB - A novel method for selection of transgenic plants utilizing the sweet pepper ( Capsicum annuum L.) ferredoxin-like protein ( pflp) gene as selection marker and Erwinia carotovora as the selection agent has been developed. An expression vector containing a pflp cDNA driven by a cauliflower mosaic virus 35S promoter was successfully transformed into protocorm-like bodies of Oncidium orchid by Agrobacterium tumefaciens and particle bombardment, respectively. Erwinia carotovora was used as a selection agent to screen transformants, thereby obtaining transgenic plants without the use of an antibiotic selection agent. A total of 32 independent transgenic orchid lines were obtained, out of which 9 transgenic lines (beta-glucuronidase positive) were randomly selected and confirmed by Southern and northern blot analyses. The transgenic orchid plants showed enhanced resistance to E. carotovora, even when the entire plant was challenged with the pathogen. Our results suggest the novel use of the pflp gene as a resistance selection marker in plant genetic engineering strategies. In the future, the use of the pflp gene as a selection marker may facilitate the use of smaller gene constructs due to removal of bulky antibiotic selection and reporter genes. These constructs can then be used to incorporate additional genes of choice. PMID- 12721850 TI - Allelic variation and differential expression of methionine-rich delta-zeins in maize inbred lines B73 and W23a1. AB - The sulfur-amino-acid-rich delta-zeins of maize ( Zea mays L.) are represented by 18-kDa and 10-kDa proteins. We have cloned a novel 11-kDa methionine-rich delta zein from developing endosperm of the inbred line W23a1. The nucleotide sequence of this new delta-zein is identical to the published 10-kDa delta-zein, except for an insertion of 18 nucleotides between +316 and +333 bp from the translation start site. Antibodies raised against the recombinant 18-kDa delta-zein recognized both the 18-kDa and 10-kDa delta-zein from total seed protein extracts of different maize inbred lines. Western blot analysis revealed differences in the levels of the delta-zeins in different inbred lines and some of the inbred lines lacked either the 10-kDa or the 18-kDa delta-zeins. Northern blot analysis revealed temporal differences in the RNA transcript levels of the 11-kDa and 18 kDa delta-zeins between B73 and W23a1. Such differences were not evident on Western blot analysis where similar protein accumulation profiles were seen for both lines. Immunostaining of paraffin sections of developing maize endosperm with the 18-kDa delta-zein antibodies revealed specific labeling of protein bodies found in the first few starchy layers from the aleurone layer. Electron microscopic observation of thin-sections of B73 and W23a1 endosperm cells confirmed the presence of recently discovered novel, vacuole-like structures in these inbred lines. Immunogold labeling studies revealed that the delta-zeins were localized in the endoplasmic-reticulum-derived protein bodies and showed no preferential gold particle labeling over either the light or electron-dense material found in these protein bodies. PMID- 12721851 TI - Enhanced resistance to Phytophthora infestans and Alternaria solani in leaves and tubers, respectively, of potato plants with decreased activity of the plastidic ATP/ADP transporter. AB - Recently, it has been reported that tubers of transgenic potato ( Solanum tuberosum L.) plants with decreased activity of the plastidic ATP/ADP transporter (AATP1) contain less starch, despite having an increased glucose level [P. Geigenberger et al. (2001) Plant Physiol 125:1667-1678]. The metabolic alterations correlated with enhanced resistance to the bacterium Erwinia carotovora. Here it is shown that transgenic potato tubers, possessing less starch yet increased glucose levels due to the expression of a cytoplasm localized yeast invertase, exhibit drastic susceptibility to E. carotovora. In addition, it is demonstrated that AATP1 anti-sense tubers show an increased capacity to ward off the pathogenic fungus Alternaria solani. In contrast to AATP1 anti-sense tubers, the corresponding leaf tissue does not show changes in carbohydrate accumulation. However, upon elicitor treatment, AATP1 anti-sense leaves possess an increased capacity to release H(2)O(2) and activate various defence-related genes, reactions that are associated with substantially delayed appearance of disease symptoms caused by Phytophthora infestans. Grafting experiments between AATP1 anti-sense plants and wild-type plants indicate the presence of a signal that is generated in AATP1 rootstocks and primes wild-type scions for potentiated activation of cellular defence responses in leaves. Together, the results suggest that (i) the enhanced pathogen tolerance of AATP1 anti-sense tubers is not due to "high sugar resistance", (ii) the increased disease resistance of AATP1 anti-sense tubers is effective against different types of pathogen and (iii) a systemic signal induced by antisensing the plastidic ATP/ADP transporter in potato tubers confers increased resistance to pathogens. PMID- 12721852 TI - Suborganellar localisation and effect of light on Helianthus tuberosus chloroplast transglutaminases and their substrates. AB - The light stimulation of transglutaminase (TGase EC 2.3.2.13) activity was verified by incubating isolated chloroplasts of Helianthus tuberosus L. continuously or for alternate periods of light or dark (light/dark and dark/light). The first 10 min of incubation always represented the critical period. Light-harvesting complexes of photosystem II (LHCII) were more intensely labelled by (14)C-polyamines under light and light/dark than under dark and dark/light conditions. Chloroplasts were fractionated into thylakoid- and stroma enriched fractions in which multiple TGase forms and substrates were found. Antibodies against TGase recognised 58- and 24-kDa bands in thylakoids and a 150 kDa band in the stroma. The latter, and its 150-kDa fraction, catalysed the conjugation of 14C-polyamines to Rubisco. In both fractions (thylakoid-pre and stroma-pre) the analysis of polyamine glutamyl derivatives showed a significant light-affected conjugation of polyamines to endogenous proteins. Alternatively, entire chloroplasts were incubated and afterwards their sub-fractions were isolated (thylakoid-post and stroma-post). The PSII and LHCII complexes were more intensely immunodetected in thylakoid-post than in thylakoid-pre, especially under dark conditions. Conversely, the conjugation of polyamines to thylakoid proteins was clearly light-stimulated in thylakoid-post, and much less in thylakoid-pre. Stroma-pre proteins were poorly polyamine-conjugated and not light affected; on the contrary, stroma-post proteins were much more polyamine-modified and strongly light-stimulated. Thus, the light-activated conjugation depends mainly on the presence of the thylakoid fraction during the assay. The protective effect on chloroplasts under photo-damage, stress or senescence conditions attributed in the literature to free polyamines is discussed with regard to the occurrence of polyamine conjugates catalysed by TGases. PMID- 12721853 TI - Cinnamic acid 4-hydroxylase from cell cultures of the hornwort Anthoceros agrestis. AB - Cinnamic acid 4-hydroxylase (EC 1.14.13.11), a cytochrome P450-dependent hydroxylase was for the first time characterized from a hornwort, Anthoceros agrestis Paton (Anthocerotaceae). In suspension cultures of A. agrestis up to 5% of the dry weight was accumulated as rosmarinic acid, a natural product commonly known from higher plants (e.g. species of the Lamiaceae and Boraginaceae). Cinnamic acid 4-hydroxylase is involved in the biosynthesis of rosmarinic acid. The participation of cytochrome P450 was demonstrated by the inhibition of hydroxylase activity by cytochrome c and the inhibition of cinnamic acid hydroxylation in a CO-containing atmosphere, which is partially released by illumination with blue light. The apparent K(m) values were determined to be at 60 microM and 5 microM for NADPH and cinnamic acid, respectively. A comparatively high hydroxylation activity was seen with NADH as electron donor. While the hydroxylase activity with NADPH was strongly inhibited by the competitive electron acceptor cytochrome c, the activity with NADH was less susceptible, indicating the possibility of different electron-transfer pathways. PMID- 12721854 TI - Isolation and characterisation of Chlamydomonas reinhardtii mutants with an impaired CO2-concentrating mechanism. AB - In order to identify new genes involved in the carbon-concentrating mechanism of Chlamydomonas reinhardtii (Dangeard), high-CO2-requiring mutants were isolated by insertional mutagenesis after transformation of strain CC1618 with a plasmid carrying Arg7 as a selectable marker gene. Six mutants were classified by their growth behaviour under ambient CO2, the affinity of the CO2-concentrating mechanism for inorganic carbon and the expression of known low-CO2-inducible proteins. The mass-spectrometric measurement of carbonic anhydrase activity and CO2/HCO3- transport revealed that four of the mutants are unable to induce a high affinity carbon-concentrating mechanism. The expression of various carbonic anhydrases and chloroplast inner envelope polypeptides was examined with Western Blots. While three high-CO2-requiring mutants showed abnormal expression patterns, one matched the wild type. With Southern blots the total number and structure of the insertion events were determined to select possible candidates for plasmid recovery. Abnormal structures of thylakoid lamellae traversing the pyrenoid were detected by electron microscopy in some of the high-CO2-requiring mutants. Our characterisations of the insertionally generated mutants revealed phenotypes that have not been published before and therefore might be useful tools to obtain new insights on the molecular background of the CO2-concentrating mechanism and its regulation. PMID- 12721855 TI - Purification and characterization of pyrophosphate- and ATP-dependent phosphofructokinases from banana fruit. AB - Pyrophosphate-dependent phosphofructokinase (PFP; EC 2.7.1.90) and two isoforms of ATP-dependent phosphofructokinase (PFK I and PFK II; EC 2.7.1.11) from ripened banana ( Musa cavendishii L. cv. Cavendish) fruits were resolved via hydrophobic interaction fast protein liquid chromatography (FPLC), and further purified using anion-exchange and gel filtration FPLC. PFP was purified 1,158-fold to a final specific activity of 13.9 micromol fructose 1,6-bisphosphate produced (mg protein)(-1) x min(-1). Gel filtration FPLC and immunoblot analyses indicated that this PFP exists as a 490-kDa heterooctomer composed of equal amounts of 66- (alpha) and 60-kDa (beta) subunits. PFP displayed hyperbolic saturation kinetics for fructose 6-phosphate (Fru 6-P), PPi, fructose 1,6-bisphosphate, and Pi ( K(m) values = 32, 9.7, 25, and 410 microM, respectively) in the presence of saturating (5 microM) fructose 2,6-bisphosphate, which elicited a 24-fold enhancement of glycolytic PFP activity ( K(a)=8 nM). PFK I and PFK II were each purified about 350-fold to final specific activities of 5.5-6.0 micromol fructose 1,6 bisphosphate produced (mg protein)(-1) x min(-1). Analytical gel filtration yielded respective native molecular masses of 210 and 160 kDa for PFK I and PFK II. Several properties of PFK I and PFK II were consistent with their respective designation as plastid and cytosolic PFK isozymes. PFK I and PFK II exhibited: (i) pH optima of 8.0 and 7.3, respectively; (ii) hyperbolic saturation kinetics for ATP ( K(m)=34 and 21 microM, respectively); and (iii) sigmoidal saturation kinetics for Fru 6-P ( S0.5=540 and 90 microM, respectively). Allosteric effects of phospho enolpyruvate (PEP) and Pi on the activities of PFP, PFK I, and PFK II were characterized. Increasing concentrations of PEP or Pi progressively disrupted fructose 2,6-bisphosphate binding by PFP. PEP potently inhibited PFK I and to a lesser extent PFK II ( I50=2.3 and 900 microM, respectively), while Pi activated PFK I by reducing its sensitivity to PEP inhibition. Our results are consistent with: (i) the respiratory climacteric being regulated by fine (allosteric) control of pre-existing enzymes; and (ii) primary and secondary glycolytic flux control being exerted at the levels of PEP and Fru 6-P metabolism, respectively. PMID- 12721857 TI - Ethylene regulates the timing of anther dehiscence in tobacco. AB - We investigated the involvement of ethylene signaling in the development of the reproductive structures in tobacco ( Nicotiana tabacum L.) by studying flowers that were insensitive to ethylene. Ethylene-insensitivity was generated either by expression of the mutant etr1-1 ethylene-receptor allele from Arabidopsis thaliana or by treatment with the ethylene-perception inhibitor 1 methylcyclopropene (MCP). Development of ovaries and ovules was unaffected by ethylene-insensitivity. Anther development was also unaffected, but the final event of dehiscence was delayed and was no longer synchronous with flower opening. We showed that in these anthers degeneration of the stomium cells and dehydration were delayed. In addition, we found that MCP-treatment of detached flowers and isolated, almost mature anthers delayed dehiscence whereas ethylene treatment accelerated dehiscence. This indicated that ethylene has a direct effect on a process that takes place in the anthers just before dehiscence. Because a similar function has been described for jasmonic acid in Arabidopsis, we suggest that ethylene acts similarly to or perhaps even in concurrence with jasmonic acid as a signaling molecule controlling the processes that lead to anther dehiscence in tobacco. PMID- 12721856 TI - Phosphatidic acid produced by phospholipase D is required for tobacco pollen tube growth. AB - Phospholipase D (PLD) and its product phosphatidic acid (PA) are involved in a number of signalling pathways regulating cell proliferation, membrane vesicle trafficking and defence responses in eukaryotic cells. Here we report that PLD and PA have a role in the process of polarised plant cell expansion as represented by pollen tube growth. Both phosphatidylinositol-4,5-bisphosphate dependent and independent PLD activities were identified in pollen tube extracts, and activity levels during pollen tube germination and growth were measured. PLD mediated PA production in vivo can be blocked by primary alcohols, which serve as a substrate for the transphosphatidylation reaction. Both pollen germination and tube growth are stopped in the presence 0.5% 1-butanol, whereas secondary and tertiary isomers do not show any effect. This inhibition could be overcome by addition of exogenous PA-containing liposomes. In the absence of n-butanol, addition of a micromolar concentration of PA specifically stimulates pollen germination and tube elongation. Furthermore, a recently established link between PLD and microtubule dynamics was supported by taxol-mediated partial rescue of the 1-butanol-inhibited pollen tubes. The potential signalling role for PLD derived PA in plant cell expansion is discussed. PMID- 12721858 TI - Arabidopsis glucosyltransferases with activities toward both endogenous and xenobiotic substrates. AB - Arabidopsis thaliana Heynh. harbors UDP-glucose-dependent glucosyltransferase (UGT; EC 2.4.1.-) activities that are able to glucosylate xenobiotic substrates as a crucial step in their detoxification, similar to other plants. However, it has remained elusive whether side-activities of UGTs acting on endogenous substrates could account for that property. Therefore, seven recombinantly expressed A. thaliana enzymes were tested using the phytotoxic xenobiotic model compound 2,4,5-trichlorophenol (TCP) as a substrate. The enzymes were selected from the large Arabidopsis UGT gene family because their previously identified putative endogenous substrates comprised both carboxylic acid, and phenolic and aliphatic hydroxyl moieties as biochemical targets. In addition, UGT75D1, which was shown to accept the endogenous flavonoid kaempferol as a substrate, was included. All enzymes tested, except the sterol-conjugating UGT80A2, glucosylated TCP as a parallel activity. The K(m) values for TCP ranged from 0.059 to 1.25 mM. When tested at saturating concentrations of the native substrates the glucosylation of TCP by the glucose-ester-forming UGT84A1 and UGT84A2 was suppressed by p-coumaric acid and sinapic acid, respectively. In contrast, the activities of UGT72E2 and UGT75D1 toward their phenolic native substrates and the xenobiotic TCP were mutually inhibited. TCP was a competitive inhibitor of sinapyl alcohol glucosylation by UGT72E2. These overlapping in vitro activities suggest cross-talk between the detoxification of xenobiotics and endogenous metabolism at the biochemical level, depending on the presence of competing substrates and enzymes. PMID- 12721859 TI - Ionic and osmotic disruptions of the lily pollen tube oscillator: testing proposed models. AB - Two mechanisms have been proposed as the primary control of oscillating tip growth in Lilium longiflorum Thunb. pollen tubes: changes in cell wall strength (Holdaway-Clarke et al. 1997) or alternatively, changes in turgor pressure (Messerli et al. 2000). Here we have modified the ionic and osmotic concentrations of the growth medium to test predictions derived from both models. Raising the [Ca2+]o tenfold above normal reduced the amplitude of the [Ca2+]i oscillations and growth oscillations while it raised the basal [Ca2+]i and growth rate such that the average growth rate did not change. Raising the [H+] of the growth medium tenfold reversibly decreased and sometimes eliminated the [Ca2+]i and growth oscillations without changing the average growth rate. Lowering the [H+] tenfold led to irregular frequency and amplitude [Ca2+]i oscillations, reduced the average growth rate of tubes and led to cell bursting in 33% of tubes. Addition of 50 mM H+ buffer, MES, to prevent pH changes in the cell wall increased the period, amplitude and duration of both [Ca2+]i and growth oscillations. Changing the [K+]o did not markedly effect [Ca2+]i oscillations. Reducing the osmolarity of the medium led to transient large-amplitude [Ca2+]i and growth oscillations while reducing large-amplitude oscillations over long periods. In many different conditions under which growth still occurs, lily pollen tubes maintain growth oscillations, albeit with modified frequency, amplitude and duration. We conclude that modifications to both proposed models are necessary to explain oscillating growth in this system. PMID- 12721862 TI - [The olfactory bulb as a model for experimental studies]. AB - The olfactory bulb represents a model for several research works carried out in our laboratory. Several details concerning structure, cell varieties and connections have been described. Special attention should be given to the ensheathing glia which accompanies olfactory axons, it being responsible for its plasticity and regeneration, as well as intervening in glomerular formation and their specificity. The olfactory system also represents a good model for the study of certain developmental aspects of the nervous system, such as the formation of nerve tracts. The lateral olfactory tract extends into the basal telencephalon and its development is supported by the existence of pre-existing routes and several attractive or repulsive factors. Its development requires complex molecular interactions in combination with several signalling influences. Pax6 is a transcription factor required for brain development. Nasal structures and eyes are absent in the homozygous mutant mice. In this mutant we have described the presence of one particular structure which could be a prospective olfactory bulb. Specific cell markers have been used to demonstrate that this structure shows many structural characteristics of the olfactory bulb. These observations favor the hypothesis of the existence of specific patterns delimiting specific functional domains in absence of their proper afferent fibers. PMID- 12721861 TI - Salicylic acid-induced aluminum tolerance by modulation of citrate efflux from roots of Cassia tora L. AB - Aluminum-induced exudation of organic acids from roots has been proposed as a mechanism for Al tolerance in plants. To better understand the regulatory process leading to efflux of organic acids, the possible involvement of salicylic acid (SA) in regulating Al-induced citrate release in Cassia tora L. was identified. The response of citrate efflux to exogenous SA was concentration-dependent. Application of SA at 5 microM in solution containing 20 microM Al increased citrate efflux to levels 1.76-fold higher than in controls (20 microM Al alone). However, inhibition of citrate release was observed when SA concentrations increased to more than 20 microM. Increased citrate efflux due to the SA treatment was associated with decreased inhibition of root growth and Al content in root tips, suggesting that exogenous SA could confer Al tolerance by increasing citrate efflux. We also examined citrate synthase activities (EC 4.1.3.7) and citrate concentrations in root tips exposed to Al and/or SA. However, both citrate synthase activities and citrate accumulation remained unaffected. These results indicate that SA-promotion of Al-induced citrate efflux is not correlated with increase in citrate production. Total endogenous SA concentrations were measured in root tips and the SA concentrations were significantly enhanced by Al at levels of 10-50 microM. PMID- 12721860 TI - Overexpression of the sucrose transporter SoSUT1 in potato results in alterations in leaf carbon partitioning and in tuber metabolism but has little impact on tuber morphology. AB - The aim of this work was to examine the consequences of the heterologous expression of a spinach ( Spinacia oleracea L.) sucrose transporter ( SoSUT1) in potato ( Solanum tuberosum L.). Many studies have indicated that reduction of the expression of this class of sucrose transporter has deleterious effects on plant growth and development; however, until now the possibility of improving plant performance by enhancing the expression of this sucrose transporter has not been reported. With this intention we constructed a chimeric construct in which SoSUT1 was cloned in-frame with the myc epitope. We confirmed that this construct, SoSUT1m, was able to mediate sucrose transport by expression in the yeast strain SUSY7. SoSUT1m was expressed in wild-type potato in the sense orientation under the control of the cauliflower mosaic virus 35S promoter to evaluate the effect of an increased constitutive expression of a class-I sucrose transporter. We confirmed that these plants displayed expression of SoSUT1 at both the transcript and protein level and that microsomal fragments isolated from selected lines had an increased sucrose uptake capacity. Analysis of metabolism of these lines indicated that the leaves were characterised by a reduced sucrose level yet exhibited little change in photosynthetic rate. Furthermore, despite the observed increase in sugar (and reduction in amino acid) levels within the tubers, there was little change in either starch content or tuber yield in the transformants. In summary, the genetic manipulation described in this paper resulted in a shift in carbon partitioning in both leaves and tubers and an increased sucrose uptake rate in plasma-membrane vesicles isolated from these lines, but had little impact on tuber metabolism or morphology. PMID- 12721863 TI - [Bilateral subthalamic nucleus deep-brain stimulation (STN-DBS) in Parkinson's disease: initial experience in Cruces Hospital]. AB - INTRODUCTION: Clinical outcomes of Parkinson's disease patients treated for 12 months with STN-DBS were analyzed. PATIENTS ADN METHODS: Twelve patients were selected using the CAPSIT protocol criteria and placement of electrodes in the appropriate target was performed according to results of fusion image techniques and intraoperative microrecording. RESULTS: A reduction in motor UPDRS (44 %) and activities of daily living (58 %) scores during <> phases were observed. <> time with dyskinesias was reduced (86 %), while severe dyskinesias disappeared. Levodopa dosage was also lowered (44 %). Patients and caregivers showed a clear-cut benefit on quality of life (58 % and 61 % respectively). No cognitive deterioration was observed and morbidity was in the same range as that published by other teams. CONCLUSION: Bilateral STN-DBS is an effective symptomatic therapy for complicated Parkinsons disease patients. It improves the quality of life of patients and their caregivers and allows a reduction of levodopa dosage. PMID- 12721865 TI - [Cabergoline in the treatment of Parkinson's disease]. AB - Cabergoline (1-[(6-allelylergolin-8 beta-yl)carbonyl]-1-[3-(dimethylamino)propyl] 3-ethyl-urea) is a new agonist of the D2 dopaminergic receptors used in the treatment of Parkinson's disease. Cabergoline is characterized by unique pharmacologic properties, such as its long plasma half-life (about 68 hours), which allows for once a day administration. Cabergoline is well tolerated, as has been shown in several clinical trials. Based on the information available, we suggest that cabergoline produces an improvement in the symptoms of Parkinson's disease similar to those produced by other dopaminergic agonists. Cabergoline monotherapy, when used in previously untreated patients, is an appropriate option for the symptomatic treatment of Parkinson's disease. Cabergoline improves motor symptoms, delays the presentation of levodopa-induced motor complications, and diminishes the amount of levodopa required for the control of the symptoms. We suggest that cabergoline is an adequate adjuvant treatment for Parkinson' disease. There is improvement in motor symptoms (without substantially increased dyskinesias), reduced severity and duration of the wearing-off period, and diminished need for levodopa. Cabergoline can also be useful in the treatment of sleep disturbances associated with advanced Parkinson's disease such as nocturnal akinesia and dystonia. However, additional studies on cabergoline's effects in nocturnal disturbances associated with Parkinson's disease are still required. Cabergoline is a well tolerated drug. Its side effects are seen mainly in the digestive and nervous system (central and peripheral). The efficacy of cabergoline in comparison to other dopaminergic agonists should be tested in future clinical studies. PMID- 12721864 TI - [Interictal basal brain SPECT versus interictal activated brain SPECT in temporal cryptogenic epilepsy]. AB - INTRODUCTION: Interictal epileptiform activity (IEA) is a common finding in temporal epileptic patients. The nature of the IEA is not entirely understood; it may be microseizures or just a negative phenomenon expressing brain inhibition mechanisms. In order to understand how brain metabolism is during interictal activity we studied a group of patients with SPECT HMPAO during basal condition no ictal activity - and when the same patients were having interictal activity. Temporal, thalamic and cerebellar perfusion were quantified by a digital method. PATIENTS AND METHODS: Two studies were performed in 24 epileptic patients: (9/15:M/F, 36,5 14,6 years). The MRI was normal or had minimal changes. Previous EEG had showed frontotemporal foci. The patients were studied by SPECT HMPAO performed with a STARCAM 3.200 equipment, the images being obtained by high resolution collimator with circular orbit 64 images, 30 s per image. The first study was performed when there was no anomaly in the EEG (basal study). In the second study (activated study) the radioligand was injected when the EEG showed at least 10 graphoelements (spike, polispikes, or sharp waves per EEG page). RESULTS: Thalamic, temporal and cerebellar perfusion decreased significantly in the activated state compared to the basal one (p < 0.001). The statistical relationship between these three locations was studied. There was a significant relationship (p < 0.001) between thalamic, temporal, and cerebellar perfusion, both in basal state and in the activated one. The correlation index was R2 > 0.9. CONCLUSIONS: For the cases studied, temporal epilepsy is associated to a significant decrease in thalamic, temporal and cerebellar perfusion in the basal state compared to the activated one. This finding may indicate that IEA expresses a post-ictal state. PMID- 12721866 TI - [Health-related quality of life in multiple sclerosis]. AB - The clinical scales used to assess disease severity in multiple sclerosis (MS) patients share a common feature: assessment is made and interpreted by neurologists, and, therefore, someone other than the patient. Furthermore, these scales do not assess the health factors that are important components of patient's experienced quality of life. Health-related quality of life (HRQoL) instruments include several domains, which are regarded by patients as being more important determinants of their overall health states. In fact, HRQoL measurements in MS are becoming important in assessing the effect of treatment and progression of the disease. This article will provide a review of the most significant studies on HRQoL in MS. PMID- 12721867 TI - [Bilateral occipital intraparenchymatous hemorrhage]. PMID- 12721868 TI - [Traumatic dissection of the vertebral arteries]. PMID- 12721869 TI - [Megadolicobasilar, ulcerative colitis and ischemic stroke]. AB - Megadolichobasilar (MDB) is a rare arterial anomaly consisting of excessive elongation, widening and tortuosity of the basilar artery. It may be associated with different neurological disturbances, including cerebral ischemic stroke, compression of the cranial nerves, hydrocephalus, headache and vertigo. Ulcerative colitis (UC) is an inflammatory bowel disease of unknown aetiology which may be complicated by arterial or venous cerebral illness, among others neurological anomalies. We report a patient suffering from UC who presented ischemic stroke. The neurorradiological studies showed incompletely thrombosed MDB accompanied by a distal aneurysm from narrow zone. These findings advised anticoagulant treatment which leads to neurological stability. To our knowledge, it is the first report of MDB associated with UC. Although this association is probably fortuitous, we discuss both the etiopatogeny and the possibly influence of each one on the clinical picture. PMID- 12721870 TI - [Crazy laughter: first manifestation of a pontine tumour]. AB - Crazy laughter (<>) was first described in 1903 as a prodromic symptom of an ischemic stroke and was later associated with brain lesions having a different location and etiology. We describe the case of a patient with a poorly differenciated pulmonary carcinoma who presented a centropontine image consistent with metastasis, whose initial manifestation was involuntary, persistent and unmotivated laughter that preceded other clinical manifestations. We revised, on the one hand, previous cases described in the literature of pathological laughter in relationship to structural lesions, of vascular or neoplastic etiology, and, on the other, the nervous centers and pathways that control the laughter mechanism. PMID- 12721871 TI - [CADASIL: a case with clinical, radiological, histological and genetic diagnoses]. AB - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a rare inherited cerebrovascular disease. The onset of clinical symptoms occurs with migraine with aura, transient ischemic attacks, recurrent subcortical ischemic infarcts, neuropsychiatric changes reaching subcortical dementia. Brain magnetic resonance images show multiple deep cerebral infarcts in white matter and basal ganglia and diffuse leukoencephalopathy. Neuropathologic hallmark consists of deposition of small electron dense granular patches related to the basement membrane of vascular smooth muscle cells with degeneration of smooth muscle cells and media and luminal obliteration. Recently, the genetic characteristics of this disorder have been reported. Missense mutations in notch3 gene localized in chromosome 19 are involved in its pathogenesis. Only three families from Spain have been reported. Here we describe a patient with typical clinical symptoms, neuroimaging and pathology of CADASIL. C406T (Arg110Cys) mutation in notch3 gene was found. We comment on the clinical symptoms of different members of the patient's family. PMID- 12721873 TI - [Lhermitte's sign in a familial form of cerebral cavernous malformations]. PMID- 12721874 TI - [On metamorphosis and metamorphopsia: terminology and definition]. PMID- 12721872 TI - [Myastenia gravis unmasked by botulinum toxin]. PMID- 12721875 TI - [Influence of institutional factors in neurological, medical and geriatric departments on length of stay in patients with stroke]. AB - BACKGROUND: Length of hospital stay (LOS) is determined not only by medical procedures or complications but also by institutional factors. We examined the influence of various institutional factors in neurological, medical and geriatric departments on LOS in patients with ischemic stroke. METHODS: We used data of 12,410 patients from the Westphalian Stroke Register for the years 2000 and 2001. Forty-two centres including 24 neurological, 13 medical and five geriatric departments participated in the register. The register is based on a standardized data assessment, including patient-related sociodemographic and clinical items, diagnostic and treatment procedures, complications, and status at discharge. RESULTS: 7855 patients with ischemic stroke from 37 centres (median age: 73 years, 51 % female) were included in the analysis. In neurological departments, the LOS decreased with increasing numbers of stroke patients treated per centre and year, presence of a stroke unit or a rehabilitation unit. Conversely, the ratio beds to number of physicians was positively associated with LOS. In geriatric departments, a significant decrease in LOS with an increasing number of stroke cases and availability of a rehabilitation unit was also observed. In departments of medicine, no significant influence on LOS was found for the institutional factors analysed. CONCLUSIONS: Institutional factors have a significant influence on LOS in patients with stroke. In this analysis, the influence varied between the different medical specialties. Institutional factors gain importance in the management of stroke patients, when Diagnosis Related Groups (DRG) are introduced. PMID- 12721876 TI - [Cholestasis and vomiting: unusual differential diagnosis in a case of Peutz Jeghers syndrome]. AB - HISTORY AND CLINICAL FINDINGS: A 54-year-old patient with painless jaundice and vomiting had been diagnosed with a Peutz-Jeghers syndrome 20 years before. INVESTIGATIONS: The blood analysis showed a cholestatic constellation as well as increased transaminases. Sonographic, radiological, endoscopic and histological findings indicated multiple hamartomatous polyps of the Peutz-Jeghers' type in the entire small and large bowel with occlusion of the papilla of Vater and the superior gastrointestinal tract by a big polyp. TREATMENT AND COURSE: After an initial percutaneous transhepatic cholangiographic drainage, Whipple's operation and a segmental resection of small and large bowel were performed. A highly differentiated adenocarcinoma of the duodenum was found in the resected specimen. CONCLUSION: This case demonstrates the potentially severe complications of a Peutz-Jeghers syndrome that had been neglected for years. PMID- 12721877 TI - [Deep venous thrombosis--case report]. PMID- 12721878 TI - [Deep venous thrombosis--diagnostic]. PMID- 12721879 TI - [Deep venous thrombosis--treatment]. PMID- 12721882 TI - [Natriuretic peptides BNP and NT-pro-BNP--the "new troponins" for estimation of heart failure?]. PMID- 12721883 TI - [Supervision duty of the physician in ambulatory dispensation of sedatives. Decision of the Court of Appeals, Frankfurt a. M., 6/12/2002 - 1 0 56/96]. PMID- 12721884 TI - Impact of alternative approaches to accelerated measles control: experience in the African region, 1996-2002. AB - From 1996 to 2000, several African countries accelerated measles control by providing a second opportunity for measles vaccine through supplemental campaigns. Fifteen countries completed campaigns in children aged 9 months to 14 years. Seven countries completed campaigns in children aged 9-59 months. In almost all countries that conducted campaigns in children aged 9 months to 14 years, measles deaths were reduced to near zero. In six countries, near-zero measles mortality has been maintained for 4-6 years. Supplemental immunization in children <5 years old was only partially effective (range, 0-67%) in reducing mortality. Measles cases decreased by 50% when routine vaccination coverage increased from 50% to 80%. Initial measles campaigns in children aged 9 months to 14 years, follow-up campaigns in those aged 9-59 months every 3-5 years, and increased routine coverage to 80% will be needed to reduce and maintain measles deaths in African countries at near zero. PMID- 12721885 TI - The unfinished measles immunization agenda. AB - Despite achieving and sustaining global measles vaccination coverage of about 80% over the past decade, worldwide measles remains the fifth leading cause of mortality among children aged <5 years. In May 2002, the United Nations Special Session on Children endorsed the goal of reducing measles deaths by half by 2005. Countries and World Health Organization (WHO) regions that adopted aggressive measles control or elimination strategies have shown excellent results. In 2001, countries in the Americas reported an all time low of 537 confirmed measles cases. Substantial progress in measles control has also been achieved in the WHO Western Pacific Region, in seven southern African countries, and in selected countries in WHO European, Eastern Mediterranean, and Southeast Asian regions. The ongoing measles disease burden and availability of safe and effective measles mortality reduction strategies make a compelling case to complete the unfinished agenda of measles immunization. PMID- 12721886 TI - The global burden of measles in the year 2000--a model that uses country-specific indicators. AB - The estimation of the global burden of measles is challenging in the absence of reliable and comparable surveillance systems worldwide. A static model is described that enables estimation of measles morbidity, mortality, and disability for the year 2000 on the basis of country-specific information (i.e., demographic profile, vaccine coverage, and estimates of case-fatality ratios). This approach estimated a global incidence of 39.9 million measles cases, 777,000 deaths, and 28 million disability-adjusted life years. The World Health Organization regions of Africa and Southeast Asia had 70% of incident cases and 84% of measles-related deaths; 11 countries alone (Afghanistan, Burkina Faso, Democratic Republic of the Congo, Ethiopia, India, Indonesia, Niger, Nigeria, Pakistan, Somalia, Uganda) account for 66% of deaths. This approach quantifies the measles burden by considering country-specific indicators, which can be updated, permitting an assessment of country, regional, and global changes in the burden associated with measles infection. PMID- 12721888 TI - Determining measles-containing vaccine demand and supply: an imperative to support measles mortality reduction efforts. AB - Measles remains a major cause of mortality with an estimated 745,000 deaths in 2001. The timely, sustained, and uninterrupted supply of affordable vaccines is critical for global efforts to reduce measles mortality. The measles vaccine supply needs to be considered in the context of vaccine security. In 2000, the World Health Organization (WHO) and United Nations Children's Fund (UNICEF) issued a number of new recommendations for measles control that resulted in a two fold increase in the number of measles-containing vaccine (MCV) doses administered between 2000 and 2002. Any additional increments in mass campaigns must be duly planned and have time lines so that vaccine production capacities are increased to optimal levels. The cornerstone of vaccine security efforts remains at the country level. WHO and UNICEF, with major partners, will review progress on measles mortality reduction and assess the feasibility of global measles eradication. Strong collaboration by all key stakeholders will be invaluable. PMID- 12721887 TI - Experience in global measles control, 1990-2001. AB - Worldwide during the 1980s remarkable progress was made in controlling measles through increasing routine measles vaccination to nearly 80%. In 2000, an estimated 777,000 measles deaths occurred, of which 452,000 were in the African Region of the World Health Organization (WHO). In 2001, WHO and the United Nations Children's Fund published a 5-year strategic plan to reduce measles mortality by half by 2005. Strategies include providing a second opportunity for measles immunization to all children through nationwide supplementary immunization activities, increasing routine vaccination coverage, and improving surveillance with laboratory confirmation of suspected measles cases. In 2000, over 100 million children received a dose of measles vaccine through supplementary immunization activities, a number projected to increase during 2002 2005. Current systems for monitoring measles vaccination coverage and disease burden must be improved to accurately assess progress toward measles control goals. PMID- 12721889 TI - Future savings from measles eradication in industrialized countries. AB - Estimates are made of monetary savings associated with measles eradication in seven industrialized countries. Three scenarios were studied: First, changing from the present two-dose measles-mumps-rubella (MMR) immunization schedule to one-dose of MMR; second, the use of an MMR and mumps-rubella schedule; or third, continuing the present schedule. Results show that the largest savings (US $623 million) would be achieved by changing to a one-dose MMR schedule with an assumption of a 3% discount rate and measles eradication in 2010. The smallest overall savings would result from option 3, by use of a 5% discount rate and the assumption that measles eradication occurs in 2020 ($10 million). These savings are less than previously estimated for the United States, partly because of the assumption that measles vaccines will continue to be delivered in response to possible bioterrorism threats. PMID- 12721890 TI - Progress in the control of measles in Ghana, 1980-2000. AB - By review of available literature, routine surveillance data, coverage surveys, and hospital records, measles control in Ghana was assessed since vaccinations began in 1978. Nationally, measles vaccination coverage increased from 24% in 1980 to 84% in 2000. This achievement is attributed to health sector reforms that included a higher district share of the total recurrent health budget from 20% in 1996 to 42% in 1999. The budget reallocation resulted in improved access to immunization services, supply procurement, transport management, staff motivation, and information flow. On the client side, the age of the child, socioeconomic status of parents, and type of prenatal care were associated with vaccination coverage. Routine vaccination coverage of >80% has resulted in lower measles incidence, a longer interepidemic interval, and a shift in cases to older children. Ghana recently developed a strategic plan to reduce measles deaths to near zero. PMID- 12721891 TI - Failure of targeted urban supplemental measles vaccination campaigns (1997-1999) to prevent measles epidemics in Mozambique (1998-2001). AB - This study assessed the effect of urban supplemental measles vaccination campaigns (1997-1999) in Mozambique that targeted children aged 9-59 months. Reported measles cases were analyzed to the end of 2001 to determine campaign impact. Hospital inpatient data were collected in the national capital and in three provincial capitals where epidemics occurred the year after the campaigns. Measles epidemics followed campaigns in the capital city, in 4 of 9 provincial capitals, and in 39 of 126 districts. Reasons for limited campaign impact included a low proportion of urban dwellers, the geographic location of some provincial capitals, the limited target age group, and low routine and campaign coverage. Routine immunization and disease surveillance should be strengthened and campaigns must achieve >90% coverage and target wider age groups and geographic areas in order to reach a high proportion of persons susceptible to measles. PMID- 12721892 TI - An outbreak of measles in Tanzanian refugee camps. AB - From March 2000 to May 2001, four camps in Kibondo District, Tanzania, hosting refugees from Burundi reported 1062 cases of measles, a highly infectious and potentially lethal disease. Of 1062 case-patients, 225 (21%) were <9 months old, 286 (27%) were 9 months to 5 years, 324 (31%) were 6-15 years, and 227 (21%) were >/=16 years old. No deaths were reported. Although, in accordance with Sphere Project guidelines for humanitarian emergencies, camp policy was to vaccinate all new arrivals aged 6 months to 15 years against measles, 152 (72%) of 210 newly arrived refugees in this age group were unvaccinated; 143 (94%) of the 152 had lived in the camp >/=1 month before rash onset. This investigation supports Sphere Project recommendations for wide age group vaccination and suggests that in some circumstances vaccination of refugees >15 years old may be beneficial. PMID- 12721893 TI - Impact of mass measles campaigns among children less than 5 years old in Uganda. AB - In 1999-2001, a national measles control strategy was implemented in Uganda, including routine immunization and mass vaccination campaigns for children aged 6 months to 5 years. This study assesses the impact of the campaigns on measles morbidity and mortality. Measles cases reported from 1992 through 2001 were obtained from the Health Management Information System, and measles admissions and deaths were assessed in six sentinel hospitals. Measles incidence declined by 39%, measles admissions by 60%, and measles deaths by 63% in the year following the campaigns, with impact lasting 15 to 22 months. Overall, 64% of measles cases were among children <5 years of age, and 93% were among children /=5 years old. In areas with complex emergencies, measles vaccination is recommended for those aged 6 months to 12-15 years. From December 2001 to May 2002, Afghan authorities and national and international organizations targeted 1,748,829 children aged 6 months to 12 years in five provinces in central Afghanistan for measles vaccinations. Two provinces reported coverage of >90% and two >80%. Coverage in Kabul city was 62%. A subsequent cluster survey in the city found 91% coverage (95% confidence interval [CI], 0.85-0.91) among children 6-59 months and 88% (95% CI, 0.87-0.95) among those 5-12 years old. Thus, this campaign achieved acceptable coverage despite considerable obstacles. PMID- 12721913 TI - Measles and rubella in the World Health Organization European region: diversity creates challenges. AB - Since 1984, the World Health Organization (WHO) European Region has had targets for reducing the burden of a number of communicable diseases. While some countries have already met the targets for interrupting indigenous measles transmission and for reducing the incidence of congenital rubella syndrome to <1 case per 100,000 births, most have not. The cultural and economic diversity of the region present a number of challenges that must be overcome before the regional targets are met. These include social factors, political will, economic costs associated with supplementary campaigns, and more effective communication with health professionals and the public on the benefits and risks associated with immunization. Most WHO European Region member states are expected to use combined measles-mumps-rubella vaccine within the next 5 years. Consultation within the region is occurring on a strategic plan to meet the targets by 2010. PMID- 12721914 TI - The elimination of indigenous measles transmission in England and Wales. AB - Following a school-based measles-rubella vaccination campaign in November 1994, enhanced surveillance of measles, including IgM antibody testing of oral fluid from clinically diagnosed case-patients, was introduced in England and Wales. Between 1995 and 2001, 665 cases of measles were confirmed, including 371 (56%) confirmed only by IgM detection in oral fluid. Two hundred thirty-nine cases (36%) were sporadic and 426 (64%) were associated with 61 clusters. Fifty-four (23%) of the 239 sporadic cases and 26 (43%) of the 61 clusters were associated with a probable or possible importation of infection from overseas, and a wide variety of genotypes were identified in each calendar year. The effective reproduction number for measles over the period was estimated to be below 0.7. These data suggest that most measles in the UK is acquired following limited transmission from an imported infection, and they confirm that measles elimination has been achieved and sustained over this period. PMID- 12721915 TI - Progress toward measles elimination in Germany. AB - While the former East Germany (FEG) achieved a reduction of measles incidence to <1 case per 100,000 population before reunification in 1990, the former West Germany (FWG) experienced significant measles morbidity. In 2001, according to statutory surveillance data, the incidence of measles was still higher in FWG than in FEG (8.7 vs. 0.7 cases/100,000 population). This article describes the development of the vaccination strategies in FEG and FWG, vaccination coverage, results of seroprevalence studies, measles surveillance in Germany, the epidemiology of a recent outbreak, and the role of laboratory diagnosis for measles control in Germany. Recent establishment of comprehensive nationwide surveillance and prevention programs to attain higher vaccine coverage have led to a decrease in measles incidence. However, further improvement of age appropriate vaccine coverage and closure of immunity gaps in school-age children are necessary to eliminate measles in Germany. PMID- 12721917 TI - Measles-rubella mass immunization campaign in Albania, November 2000. AB - In 2000, Albania resolved to eliminate measles by 2007 by use of a four-step program: by conducting a "catch-up" vaccination campaign for all children aged 1 14 years, achieving and sustaining high coverage (>/=95%) among children aged 1 year with the first dose of a measles-containing vaccine, by introducing a routine second dose of measles-containing vaccine for children at age 5 years, and by improving measles surveillance. This catch-up campaign took place in November 2000: 867,000 doses of measles-rubella vaccine were administered for an estimated coverage of 99%. In all, 231 campaign-related adverse events were reported: syncope, 206; allergic reactions, 10; fever, 8; encephalitis/encephalopathy, 2; and aseptic meningitis, seizures, Guillain-Barre syndrome, anaphylaxis, and arthralgia, 1 each. All resolved without sequelae. This report describes the status of measles and rubella/congenital rubella syndrome control in Albania before 2000 and reports on implementation of the catch-up campaign. PMID- 12721916 TI - Progress toward measles elimination in Romania after a mass vaccination campaign and implementation of enhanced measles surveillance. AB - In response to an outbreak of >33,000 measles cases in 1996-1998 and to prevent an outbreak predicted for 2002, Romania conducted a nationwide measles-rubella vaccination campaign in October 1998. Some 2.1 million children aged 7-18 years were vaccinated. Data from national surveillance and seroprevalence studies conducted in three districts were used to assess the campaign and status of measles control. Surveillance data showed a dramatic drop in measles despite enhanced surveillance starting in October 1999. From October 1999 to December 2001, 400 suspected measles cases were reported, down from about 5000 cases annually in non-outbreak years. Only 29 (8%) of 386 cases with specimens were laboratory confirmed; 14 were clinically confirmed. Seroprevalence estimates showed high measles antibody levels before (92.9%) and after (94.4%) the campaign. The low number of laboratory-confirmed cases and high population immunity suggest that interruption of indigenous measles virus transmission is a real possibility for Romania. PMID- 12721918 TI - Measles epidemiology and elimination strategies in Turkey. AB - In Turkey, 15,000-30,000 measles cases have been reported annually since the 1990s. Epidemics occur every 3-4 years, and >/=90% of cases are <15 years old. The high incidence is due to inadequate vaccination coverage (nationally 84% in 2001) and immunity provided by the first dose of vaccine administered at age 9 months. The second dose, which has been recommended for first grade students since 1998, has been insufficient to provide the herd immunity necessary to control measles. The Ministry of Health launched a comprehensive program for 2002 2010 targeting measles elimination. This plan calls for a national vaccination campaign among all children aged 9 months to 14 years and routine two-dose vaccination coverage will be increased to >/=95% or follow-up campaigns will be conducted in areas not achieving high coverage levels. Also, all military recruits in 2002-2009 will be vaccinated and case-based, laboratory supported surveillance will be initiated. PMID- 12721919 TI - Investigation of a rubella outbreak in Kyrgyzstan in 2001: implications for an integrated approach to measles elimination and prevention of congenital rubella syndrome. AB - In 1999, the Ministry of Health of Kyrgyzstan adopted the goal of measles elimination. This opportunity was used to launch a rubella and congenital rubella syndrome prevention program. Between January and August 2001, a rubella outbreak occurred in Bishkek City and Chui Oblast. Rubella surveillance data were reviewed for Kyrgyzstan (1981-2000) and rubella case-patient and laboratory information from Bishkek City and Chui Oblast during the outbreak. The data suggest that rubella is endemic in Kyrgyzstan with periodic epidemics every 3-5 years. From January to August 2001, 1936 rubella case-patients were reported from Bishkek City and Chui Oblast; 242 were tested and 176 (73%) were laboratory confirmed. Most case-patients were 3-14 years old. However, the incidence rate per 100,000 among persons aged 15-35 years increased >/=40-fold from 1 in 2000 to 41 in 2001. These findings highlight the importance of introducing rubella-containing vaccine in conjunction with measles elimination activities. PMID- 12721920 TI - Measles outbreak in Sri Lanka, 1999-2000. AB - A large measles outbreak occurred in Sri Lanka from October 1999 through June 2000 following a period of low incidence. During this period, >15,000 suspected cases were reported to the central Epidemiological Unit of the Department of Health Services. The outbreak was investigated through review of surveillance data for 1999-2000, which were compared with previous surveillance data. Among the clinically confirmed cases, the highest morbidity rate (114/100,000 population) was observed among children <9 months of age. Nearly 54% of the cases were among persons >/=15 years old, and this percentage had increased over the previous decade. Forty percent of cases had a history of measles vaccination. There were 5 deaths, giving a case-fatality rate of 0.1%. Action taken during the outbreak and plans for future outbreak prevention strategies were also evaluated. PMID- 12721921 TI - Accelerated measles control in the Western Pacific region. AB - By the 1990s, an immunization program in the western Pacific had dramatically reduced measles morbidity and mortality. Building on the region's successful elimination of polio, several countries and areas achieved or are close to measles elimination, thus showing the potential for global eradication. The diverse challenges for measles control in different parts of the region have produced lessons that will help with future control, including the need for surveillance of sufficient standard to guide and monitor progress. A group of experts recognized both the potential and the challenges of the measles immunization program and proposed regional elimination as the appropriate disease control target for the region. No date was recommended for its achievement. If progress continues at the present rate, the western Pacific region should soon be able to set a target date for measles elimination. PMID- 12721922 TI - Progress in accelerated measles control in the People's Republic of China, 1991 2000. AB - Measles incidence decreased dramatically following widespread use of measles vaccine in China in 1965. To evaluate continued progress in accelerated measles control, data on measles cases reported to the National Notifiable Disease Reporting System during 1991 to 2000 were analyzed. From 1991-1995 to 1996-2000, average annual measles incidence decreased from 9.0 to 5.7 cases per 100,000 population, mortality rates fell from <0.3 to 0.1 deaths per million population, and the percentage of China's total population residing in provinces with a measles incidence of <2 cases per 100,000 population and having a measles elimination goal increased from 21% to 29%. Incidence rates were highest in western provinces and in infants and young children. Additional attention must be focused on western provinces and toward ensuring that all infants are immunized. Achieving high routine two-dose coverage with measles vaccine and enforcing school entry requirements may be highly effective strategies to support further gains in measles control. PMID- 12721923 TI - Active case-based surveillance for measles in China: lessons learned from Shandong and Henan provinces. AB - To identify issues relevant to nationwide implementation, a project was conducted during 1999-2001 to support and evaluate the development of a case-based measles surveillance system (MSS) in Shandong and Henan provinces, China. The performance of MSS surveillance and the descriptive characteristics of reported measles cases and outbreaks were analyzed. Of the 5782 suspected cases in 2001, 85% were investigated and 66% had serologic results. In all, 39% of cases were confirmed, 36% were compatible, and 25% were discarded; 81% of outbreaks identified involved <15 cases. In all, 15% of cases were temporary (floating) residents. The MSS was useful in monitoring the impact of measles control activities. Standardized laboratory quality-assurance activities and indicators should be developed while the system is still in the early stages of implementation. PMID- 12721924 TI - Development of the Global Measles Laboratory Network. AB - The routine reporting of suspected measles cases and laboratory testing of samples from these cases is the backbone of measles surveillance. The Global Measles Laboratory Network (GMLN) has developed standards for laboratory confirmation of measles and provides training resources for staff of network laboratories, reference materials and expertise for the development and quality control of testing procedures, and accurate information for the Measles Mortality Reduction and Regional Elimination Initiative. The GMLN was developed along the lines of the successful Global Polio Laboratory Network, and much of the polio laboratory infrastructure was utilized for measles. The GMLN has developed as countries focus on measles control activities following successful eradication of polio. Currently more than 100 laboratories are part of the global network and follow standardized testing and reporting procedures. A comprehensive laboratory accreditation process will be introduced in 2002 with six quality assurance and performance indicators. PMID- 12721925 TI - Update on the global distribution of genotypes of wild type measles viruses. AB - Molecular characterization of measles viruses is an important component of measles surveillance because these studies enhance our ability to identify the source and transmission pathways of the virus. Molecular surveillance is most beneficial when it is possible to observe the change in virus genotypes over time in a particular region. Such information can help to document the interruption of transmission of measles virus and thus provide an important method for assessing the effectiveness of vaccination programs. It is recommended that virus surveillance be conducted during all phases of measles control and be expanded to give an accurate description of the global distribution of measles genotypes. This review provides updated information on the circulation patterns of measles genotypes and examples of the utility of virologic surveillance. PMID- 12721926 TI - Limited diversity of measles field isolates after a national immunization day in Burkina Faso: progress from endemic to epidemic transmission? AB - Despite recent National Immunization Days in Burkina Faso, the rural province of Houet reported >400 measles cases in 2001 (82% not vaccinated). Phylogenetic analysis of 58 measles virus field isolates plus the first sequences from the Democratic Republic of the Congo and the Republic of Congo are reported. All viruses were genotype B3, which is common in the region. In Houet, there were two geographically confined genetic variants, suggesting two independent importation events. Strain diversity in Houet (1.5%) and the Congos was limited in comparison with Ibadan, Nigeria (4.6%), where measles is endemic. Strain variability, assessed by heteroduplex mobility assay, confirmed these findings. Despite large local pools of susceptible persons even after several rounds of vaccination, the limited strain diversity suggests that parts of rural Burkina Faso may be moving from an endemic to an epidemic transmission pattern of measles virus. PMID- 12721927 TI - The challenges and strategies for laboratory diagnosis of measles in an international setting. AB - Serum-based measles-specific IgM EIAs are the recommended laboratory assays for diagnosis of acute measles infections and appear to be sufficient for measles control programs. However, serum samples are not ideal for molecular characterization of measles virus. Although neither laboratory nor field-based diagnostic tests that rival the EIAs have been developed, laboratory surveillance could be improved if specimen collection were simplified. Ideally the collection method should be noninvasive, have no requirement for a cold chain, and/or have no requirement for technically sophisticated equipment. Two alternative specimen collection technologies appear promising and can be used for both diagnostics and for collecting pertinent genotyping information: oral fluid and filter paper collection methods. These methods are compared along with their respective utilities in supporting measles diagnosis and strain surveillance. PMID- 12721928 TI - Monitoring vaccine safety during measles mass immunization campaigns: clinical and programmatic issues. AB - In the planning and implementation of mass immunization campaigns, vaccine delivery has always been a priority. However, safety issues have gained increasingly more attention and grown in importance, and campaign planners must now take them into prime consideration. The World Health Organization has released guidelines to assist with the design and implementation of safety surveillance systems, primarily for developing countries, and these include a new monograph for measles mass campaigns. Experience in the past decade with mass campaigns (primarily in developed countries) shows that measles vaccine performs in these settings as anticipated from pre- and post-licensure studies. Serious adverse events are rare, even under the increased scrutiny extended during a campaign. The experience in developing country settings is growing. The implementation of safety surveillance for mass campaigns offers a unique opportunity for countries to avoid crisis situations and to begin vaccine safety monitoring in routine immunization programs. PMID- 12721929 TI - Ensuring injection safety during measles immunization campaigns: more than auto disable syringes and safety boxes. AB - Measles immunization campaigns are effective elements of a comprehensive strategy for preventing measles cases and deaths. However, if immunizations are not properly administered or if immunization waste products are not safely managed, there is the potential to transmit bloodborne pathogens (e.g., human immunodeficiency virus and hepatitis B and hepatitis C). A safe injection can be defined as one that results in no harm to the recipient, the vaccinator, and the surrounding community. Proper equipment, such as the exclusive use of auto disable syringes and safety boxes, is necessary, but these alone are not sufficient to ensure injection safety in immunization campaigns. Equally important are careful planning and managerial activities that include policy and strategy development, financing, budgeting, logistics, training, supervision, and monitoring. The key elements that must be in place to ensure injection safety in measles immunization campaigns are outlined. PMID- 12721930 TI - How does herpes simplex virus type 2 influence human immunodeficiency virus infection and pathogenesis? PMID- 12721931 TI - Recent herpes simplex virus type 2 infection and the risk of human immunodeficiency virus type 1 acquisition in India. AB - To estimate the impact of prevalent and incident herpes simplex virus type 2 (HSV 2) infection on the acquisition of human immunodeficiency virus type 1 (HIV-1), stored serum samples from a cohort of 2732 HIV-1-seronegative patients attending 3 sexually transmitted infection clinics and 1 reproductive tract infection clinic in Pune, India, were screened for HSV-2-specific antibodies. Incident HSV 2 infection was defined serologically as "recent" if a negative result of testing for HSV-2 could be documented within the previous 6 months or "remote" if >6 months had elapsed since the last negative test result. The prevalence of HSV-2 at enrollment was 43%. The HSV-2 incidence was 11.4 cases/100 person-years, and the HIV-1 incidence was 5.8 cases/100 person-years. The adjusted hazard ratios of HIV-1 acquisition from exposure to HSV-2 infection were 1.67 for prevalent HSV-2, 1.92 for remote incident HSV-2, and 3.81 for recent incident HSV-2. Recent incident HSV-2 infection was associated with the highest risk of HIV-1 in this study, which suggests that prevention of HSV-2 infection may reduce the risk of HIV-1 acquisition. PMID- 12721932 TI - Human immunodeficiency virus type 1 infection of human uterine epithelial cells: viral shedding and cell contact-mediated infectivity. AB - We examined the mechanism of human immunodeficiency virus (HIV) type 1 infection of human uterine epithelial cells to gain a clearer understanding of the events by which HIV-1 infects cells within the female reproductive tract. We demonstrated that these cells can be productively infected by HIV-1 and that infection is associated with viral RNA reverse transcription, DNA transcription, and secretion of infectious virus. Levels of viral DNA and secreted virus decreased gradually after infection. Moreover, virus released by the uterine epithelial cells shortly after infection was able to infect human T cell lines, but virus released later did not. In contrast, human CD4(+) T cell lines were infected after cocultivation with epithelial cells at both early and late stages of infection. These data demonstrated that HIV-1 infects human epithelial cells of upper reproductive tract origin and that productive viral infection of epithelial cells may be an important mechanism of transmission of HIV-1 infection in women. PMID- 12721933 TI - T cell activation is associated with lower CD4+ T cell gains in human immunodeficiency virus-infected patients with sustained viral suppression during antiretroviral therapy. AB - Although T cell activation is associated with disease progression in untreated human immunodeficiency virus type 1 (HIV-1) infection, its significance in antiretroviral-treated patients is unknown. Activated (CD38(+)HLA-DR(+)) T cell counts were measured in 99 HIV-infected adults who had maintained a plasma HIV RNA level or=1 blood sample positive for EBV. Levels of EBV varied widely. CMV shedding occurred infrequently but occurred more commonly in younger individuals (P<.03). JCV and BKV virurias were 46.7% and 0%, respectively. JCV shedding was age dependent and occurred commonly in individuals >or=40 years old (P<.03). Seasonal variation was observed in shedding of EBV and JCV, but there was no correlation among shedding of EBV, CMV, and JCV (P>.50). Thus, adults independently control persistent viruses, which display discordant, sporadic reactivations. PMID- 12721939 TI - Phylogenetic analysis of rubella virus isolated during a period of epidemic transmission in Italy, 1991-1997. AB - To study the molecular epidemiology of rubella virus during endemic transmission, phylogenetic analysis of the nucleotide sequence of the E1 gene was done with 31 isolates collected in northern Italy during 1991-1997, a period spanning 3 epidemics. The viruses segregated into distinct genotypic strains. Cocirculation of genotypic strains was detected; however, each epidemic was associated with specific strains, and strain displacement occurred concomitantly with each epidemic. Although most of the viruses from Italy belonged to rubella genotype I and many were related to viruses isolated concurrently in other European countries, 3 viruses belonged to rubella genotype II, which previously had been isolated only in Asia. Thus, intercontinental importation of viruses also occurred. PMID- 12721940 TI - Protection against group A streptococcus by immunization with J8-diphtheria toxoid: contribution of J8- and diphtheria toxoid-specific antibodies to protection. AB - A conformationally constrained, minimally conserved peptide from the M protein of group A streptococcus (GAS) has been defined. It consists of 12 amino acids from the C-repeat region within a non-M protein helix-forming sequence and is referred to as "J8." Here, we investigate the immunogenicity of a J8-diphtheria toxoid (DT) conjugate adjuvanted with the human-compatible adjuvants, SBAS2 and alum, and demonstrate that it is capable of inducing opsonic antibodies and can protect outbred mice from virulent challenge. In a range of experiments, protection correlated with the titer of J8-specific antibodies and not with the induction of J8-specific T cells. However, DT-specific antibodies (as well as J8-specific antibodies) were shown to stain the surface of fixed GAS and to be capable of opsonizing live organisms. DT may be an ideal carrier protein for J8 and other GAS peptides for GAS vaccines. PMID- 12721941 TI - Severe inflammation and reduced bacteria load in murine helicobacter infection caused by lack of phagocyte oxidase activity. AB - The vaccine-induced immune mechanisms that protect against Helicobacter pylori infection in the mouse model have not been identified. This study investigated the contribution of reactive oxygen and nitrogen intermediates to Helicobacter pathogenesis and immunity. Mice deficient in nicotinamide-adenine dinucleotide phosphate oxidase activity (gp91(phox-/-)), nitric oxide synthase activity (NOS2( /-)), or both (gp91(phox-/-)/NOS2(-/-)) were infected with Helicobacter organisms and evaluated for inflammation and bacteria load. Infection of all 3 transgenic strains resulted in significantly more inflammation than found in infected C57BL/6 wild-type mice. However, only gp91(phox-/-) and gp91(phox-/-)/NOS2(-/-) mice had significantly reduced numbers of infected gastric glands. Intranasal immunization of NOS2(-/-) or gp91(phox-/-)/NOS2(-/-) mice against H. pylori resulted in protective immunity comparable to that seen in C57BL/6 control mice. Therefore, reactive oxygen species may play a role in limiting the inflammatory response associated with H. pylori infection of the gastric mucosa but may also limit the host's ability to eradicate Helicobacter organisms. PMID- 12721942 TI - Genetic basis for nongroupable Neisseria meningitidis. AB - Nongroupable Neisseria meningitidis may constitute one-third or more of meningococcal isolates recovered from the nasopharynx of human carriers. The genetic basis for nongroupability was determined in isolates obtained from a population-based study in which 60 (30.9%) of 194 meningococcal isolates from asymptomatic carriers were not groupable. Forty-two percent of nongroupable isolates were related to serogroup Y ET-508/ST-23 clonal complex strains, the most common groupable carrier isolate from the study population. Nongroupable isolates were all rapidly killed by 10% normal human serum. The capsule loci of 6 of the ET-508/ST-23 complex strains and of 25 other genetically diverse nongroupable meningococci were studied in detail. Serogroup A or novel capsule biosynthesis genes were not found. Nongroupable isolates were genetically serogroup Y, B, or C isolates that did not express capsule but were related to groupable isolates found in the population (class I); capsule deficient because of insertion element-associated deletions of capsule biosynthesis genes (class II); or isolates that lacked all capsule genes and formed a distinct genetic cluster not associated with meningococcal disease (class III). PMID- 12721943 TI - Significant variation in serotype-specific immunogenicity of the seven-valent Streptococcus pneumoniae capsular polysaccharide-CRM197 conjugate vaccine occurs despite vigorous T cell help induced by the carrier protein. AB - Streptococcus pneumoniae capsular polysaccharides (PnPSs) induce protective antibodies but are T cell-independent type 2 antigens and are poorly immunogenic in infants. Conjugate vaccines of PnPSs linked to proteins like cross-reactive material (CRM(197)) increase PS antibody titer and elicit immunologic memory in infants. Despite being linked to an identical carrier protein, each PS component of the 7-valent PnPS-CRM(197) vaccine has different immunogenicity. To determine whether variations in conjugate-induced memory T cell responses or PnPS-specific antibody-secreting cells (ASCs) were responsible for serotype-specific differences in immunogenicity, adults were immunized with 7-valent PnPS-CRM(197), and antibody titer, vaccine component-specific CD4(+) T cell recall response, numbers of PnPS-specific ASCs, and cytokine production were measured. PnPS CRM(197) induced significantly different serotype-specific antibody titers, despite vigorous T cell recall responses to all 7 vaccine components, and production of interleukin (IL)-2, IL-5, IL-6, IL-10, and interferon-gamma. We conclude that PnPS-CRM(197) induces variable serotype-specific antibody titers, despite induction of comparable CRM(197)-specific memory T cell responses. PMID- 12721944 TI - Randomized, double-blind, controlled trial of pneumococcal vaccination in renal transplant recipients. AB - Renal transplant recipients are at increased risk for developing invasive pneumococcal disease but may have a poor response to pneumococcal polysaccharide vaccine (PPV23). For them, pneumococcal conjugate vaccine (PCV7) may be more immunogenic. Patients were given a single dose of PPV23 or PCV7 in our randomized, controlled, double-blind trial. Immunogenicity was assessed 8 weeks after vaccination by serotype-specific enzyme-linked immunosorbent assay (ELISA) and opsonophagocytic assay (OPA). Baseline demographics, renal function, time since transplantation, and immunosuppression were comparable. In the PCV7 group, the vaccine response rate was improved for serotypes 23F (P=.046) and 6B (P=.067), and mean fold increases in antibody titer were higher for serotypes 23F (P=.046) and 9V (P=.09). The response rate and mean fold increase in OPA titers were not significantly different between groups. There was a trend toward enhanced immunogenicity for PCV7 by ELISA. However, functional antibody responses were not different. PMID- 12721945 TI - Phagocytosis of Bordetella pertussis incubated with convalescent serum. AB - Convalescent serum samples were examined for the ability to promote phagocytosis of Bordetella pertussis by human neutrophils. One sample promoted phagocytosis, and 11 of the 51 samples caused a statistically significant reduction in phagocytosis, compared with that of bacteria not incubated with serum. Phagocytosis was influenced by interactions between antibodies that promoted phagocytosis and antibodies that inhibited phagocytosis. Adenylate cyclase toxin (ACT) has been shown to block phagocytosis by neutrophils. Antibodies to ACT were removed from the sample that promoted phagocytosis, by incubation with ACT-coated paramagnetic beads, and the depleted serum no longer enhanced phagocytosis. The adhesin filamentous hemagglutinin (FHA) has been shown to mediate attachment of B. pertussis to neutrophils in a way that promotes phagocytosis. Depletion of antibodies to FHA from samples that blocked phagocytosis improved phagocytosis, compared with the no-antibody control. These results suggest that antibodies to ACT can promote phagocytosis, whereas antibodies to FHA can counteract beneficial opsonins. PMID- 12721946 TI - Staphylococcus aureus menD and hemB mutants are as infective as the parent strains, but the menadione biosynthetic mutant persists within the kidney. AB - Small colony variants (SCVs) of Staphylococcus aureus were generated via mutations in menD or hemB, yielding menadione and hemin auxotrophs, respectively, and studied in the rabbit endocarditis model. No differences in the 95% infectious dose occurred between strains with regard to seeding heart valves ( approximately 10(6) cfu) or other target organs. No differences were observed between the response of the hemB mutant to oxacillin therapy and that of the parent strain in any target tissues, and significant reductions in bacterial densities were seen in all tissues (compared with untreated controls). In contrast, oxacillin therapy did not significantly reduce bacterial densities of the menD mutant in either kidney or spleen and significantly reduced densities within vegetations. These data show that SCVs are able to colonize multiple tissues in vivo and that the menD mutation provides the organism with a survival advantage during antimicrobial therapy, compared with its parent strain, in selected target tissues. PMID- 12721947 TI - Leishmania mexicana and Leishmania major: attenuation of wild-type parasites and vaccination with the attenuated lines. AB - A method for attenuation of Leishmania species by culturing in vitro under gentamicin pressure has been used successfully with Leishmania mexicana, L. major, L. infantum, and L. donovani. The attenuated lines invaded but were unable to survive within bone marrow-derived macrophages in vitro, whereas wild-type parasites survived and multiplied. The attenuated lines of L. mexicana and L. major both failed to induce cutaneous lesions in the majority of BALB/c mice over a minimum 12-week observation period after subcutaneous injection of stationary phase parasites. The attenuated line of L. mexicana retained its properties in gentamicin-free medium over 40 subcultures. The attenuated lines of L. mexicana and L. major both induced significant protection in mice against challenge with wild-type parasites. PMID- 12721948 TI - Does the diagnosis of trachoma adequately identify ocular chlamydial infection in trachoma-endemic areas? AB - We evaluated the validity of clinically determined active trachoma as a surrogate for chlamydial eye infection in 1059 children from the Egyptian arm of the Azithromycin in the Control of Trachoma study. Participants were determined to be "clinically active" if they had >or=5 follicles or intense inflammatory infiltration on the tarsal conjunctiva. Conjunctival swabs were tested using ligase chain reaction (LCR) to detect chlamydial DNA. Of clinically active children aged 1-10 years, 31% did not have infection, as determined by LCR. Conversely, 31% of infected children were not clinically active; 78% of clinically active children aged 1-5 years were infected, versus 17% of those aged 11-15 years. The proportion of clinically active children who were infected decreased from 67% before treatment to 10% 14 months after mass azithromycin treatment. Clinically active trachoma is not always a reliable marker of infection, particularly in teenagers and after treatment. PMID- 12721949 TI - Lethal synergism between influenza virus and Streptococcus pneumoniae. PMID- 12721951 TI - HLA class I alleles and cervical neoplasia. PMID- 12721952 TI - Detection of attenuated, noninfectious spirochetes in Borrelia burgdorferi infected mice, after antibiotic treatment. PMID- 12721954 TI - Variation in the HLA-G promoter region influences miscarriage rates. AB - The HLA-G gene is primarily expressed in placental cells that invade the maternal decidua during pregnancy. This gene encodes multiple isoforms that fulfill a variety of functions at the maternal-fetal interface throughout gestation. Recently, a null allele for the most abundant HLA-G isoform was associated with recurrent miscarriage in two independent studies, suggesting that reduced levels of the HLA-G1 protein may compromise successful pregnancy. We initiated the present study to determine whether other polymorphisms that could affect expression levels of HLA-G were associated with fetal loss in women participating in a 15-year prospective study of pregnancy outcome. We genotyped these subjects for 18 single-nucleotide polymorphisms in the 1,300 bp upstream of exon 1, 13 of which were identified as part of this study, as well as for an insertion/deletion (in/del) polymorphism in the 3' untranslated region. The 18 SNPs defined eight unique haplotypes. One polymorphism, -725C/G, was associated with fetal loss, with an increased risk for miscarriage in couples in which both partners carried the -725G allele, compared with couples not carrying this allele (odds ratio 2.76, 95% confidence interval 1.08-7.09; P=.035). Further, the G at nucleotide 725 creates a CpG dinucleotide, and we demonstrate that this CpG site is methylated on -725G alleles. Overall, this study identified extraordinary levels of variation in the 5'-upstream regulatory region of HLA-G and provides evidence for an association between a promoter-region SNP and fetal loss rates, further attesting to the novel features and critical role of this gene in pregnancy. PMID- 12721955 TI - Mutations of the PAX6 gene detected in patients with a variety of optic-nerve malformations. AB - The PAX6 gene is involved in ocular morphogenesis and is expressed in the developing central nervous system and numerous ocular tissues during development. PAX6 mutations have been detected in various ocular anomalies, including aniridia, Peters anomaly, corneal dystrophy, congenital cataracts, and foveal hypoplasia. However, it has not been identified in patients with optic-nerve malformations. Here, we identified novel mutations in eight pedigrees with optic nerve malformations, including coloboma, morning glory disc anomaly, optic-nerve hypoplasia/aplasia, and persistent hyperplastic primary vitreous. A functional assay demonstrated that each mutation decreased the transcriptional activation potential of PAX6 through the paired DNA-binding domain. PAX6 and PAX2 are each thought to downregulate the expression of the other. Four of the detected mutations affected PAX6-mediated transcriptional repression of the PAX2 promoter in a reporter assay. Because PAX2 gene mutations were detected in papillorenal syndrome, alternation of PAX2 function by PAX6 mutations may affect phenotypic manifestations of optic-nerve malformations. PMID- 12721958 TI - The integrated operating room system - a pathway to the or 2000 and beyond. PMID- 12721956 TI - Association of specific language impairment (SLI) to the region of 7q31. AB - FOXP2 (forkhead box P2) was the first gene characterized in which a mutation affects human speech and language abilities. A common developmental language disorder, specific language impairment (SLI), affects 6%-7% of children with normal nonverbal intelligence and has evidence of a genetic basis in familial and twin studies. FOXP2 is located on chromosome 7q31, and studies of other disorders with speech and language impairment, including autism, have found linkage to this region. In the present study, samples from children with SLI and their family members were used to study linkage and association of SLI to markers within and around FOXP2, and samples from 96 probands with SLI were directly sequenced for the mutation in exon 14 of FOXP2. No mutations were found in exon 14 of FOXP2, but strong association was found to a marker within the CFTR gene and another marker on 7q31, D7S3052, both adjacent to FOXP2, suggesting that genetic factors for regulation of common language impairment reside in the vicinity of FOXP2. PMID- 12721959 TI - The Clinical Significance of TachoComb, a Fibrin Adhesive in Sheet Form. PMID- 12721957 TI - A populationwide coalescent analysis of Icelandic matrilineal and patrilineal genealogies: evidence for a faster evolutionary rate of mtDNA lineages than Y chromosomes. AB - Historical inferences from genetic data increasingly depend on assumptions about the genealogical process that shapes the frequencies of alleles over time. Yet little is known about the structure of human genealogies over long periods of time and how they depart from expectations of standard demographic models, such as that attributed to Wright and Fisher. To obtain such information and to examine the recent evolutionary history of mtDNA and Y-chromosome haplotypes in the Icelandic gene pool, we traced the matrilineal and patrilineal ancestry of all 131,060 Icelanders born after 1972 back to two cohorts of ancestors, one born between 1848 and 1892 and the other between 1798 and 1742. This populationwide coalescent analysis of Icelandic genealogies revealed highly positively skewed distributions of descendants to ancestors, with the vast majority of potential ancestors contributing one or no descendants and a minority of ancestors contributing large numbers of descendants. The expansion and loss of matrilines and patrilines has caused considerable fluctuation in the frequencies of mtDNA and Y-chromosome haplotypes, despite a rapid population expansion in Iceland during the past 300 years. Contrary to a widespread assumption, the rate of evolution caused by this lineage-sorting process was markedly faster in matrilines (mtDNA) than in patrilines (Y chromosomes). The primary cause is a 10% shorter matrilineal generation interval. Variance in the number of offspring produced within each generation was not an important differentiating factor. We observed an intergenerational correlation in offspring number and in the length of generation intervals in the matrilineal and patrilineal genealogies, which was stronger in matrilines and thus contributes to their faster evolutionary rate. These findings may have implications for coalescent date estimates based on mtDNA and Y chromosomes. PMID- 12721960 TI - Latex allergy. PMID- 12721961 TI - Surgical staplers for vascular control of hepatic veins and portal structures in major hepatic resection. PMID- 12721963 TI - Immunotoxins for tolerance induction: preclinical results. PMID- 12721962 TI - A touch information system for healthcare worker exposure, risk, and training management. PMID- 12721964 TI - Vitamin C and the renal-failure patient: risk of oxalosis. PMID- 12721965 TI - Minimally invasive alternatives to traditional operative hepatic cryoablation: works in progress. PMID- 12721966 TI - Robotics and allied technologies in endoscopic surgery. PMID- 12721967 TI - Endoscopic ultrasound-guided real time biopsy of peri-intestinal tumors. PMID- 12721968 TI - Argon plasma coagulator: should everyone have one? PMID- 12721970 TI - Christoudias tissue approximator grasper: a step forward in minimally invasive surgery. PMID- 12721969 TI - The surgical treatment of morbid obesity. PMID- 12721971 TI - Laparoscopic nissen fundoplication using ultrasonic cutting and coagulation. PMID- 12721972 TI - Developments in hernia repair. PMID- 12721973 TI - Laparoscopic resection of a bleeding meckel s diverticulum: a case report and review of the literature. PMID- 12721974 TI - Thoracoscopic sympathectomy: a case report. PMID- 12721975 TI - Minimally invasive approaches to liver surgery. PMID- 12721976 TI - Training phantom for ultrasound with a tissue-mimicking material. PMID- 12721978 TI - Benefits and complications of laparoscopic surgery a decade after its introduction. PMID- 12721977 TI - Enhancement of upper abdominal operative field access and exposure using an operating room table accessory to achieve 62 degree head up feet down tilt (extreme reverse trendelenburg position). PMID- 12721979 TI - Complications following transanal endoscopic microsurgery. PMID- 12721980 TI - Total parenteral nutrition: techniques, complications, and prevention. PMID- 12721981 TI - Gastric endo-organ access: technique and complications. PMID- 12721983 TI - Intestinal ischemia: molecular basis of detection and prevention. PMID- 12721982 TI - Complications of mucormycosis: total gastrectomy for emphysematous gastritis. PMID- 12721984 TI - Surgical Complications: Mobile Surgery vs. Hospital Surgery. PMID- 12721985 TI - Recurrent Groin Hernia: Reduction in Complications by PerFix Hernioplasty. PMID- 12721986 TI - Off-pump coronary artery bypass surgery. PMID- 12721988 TI - Cerebral complications in cardiac surgery: the role of embolization and hypoperfusion. PMID- 12721987 TI - Cardiac valvular replacement devices: residual problems and innovative investigative technologies. AB - The advancements in cardiac valvular replacement devices over the past 25 years have left residual problems with biological and mechanical prostheses. The extensive developments were introduced to reduce or eliminate valve-related complications, namely thromboembolism, anticoagulant-related hemorrhage, and structural failure, as weil as to optimize hemodynamic performance. Residual problems persist with both biological and mechanical prostheses. Structural failure of porcine and pericardial bioprostheses persists over time with leaflet degeneration and dystrophic calcification. Thrombus formation from blood stasis and the resultant thromboembolic phenomena despite anticoagulant management remain a continuing problem with mechanical prostheses. The innovative echnologies under investigation will likely significantly improve the clinical performance of both biological and mechanical prostheses. PMID- 12721990 TI - Techniques of treatment of peritoneal endometriosis: the cavitational ultrasonic surgical aspirator. PMID- 12721989 TI - Veno-veno bypass of the inferior vena cava in trauma, tumor, and transplantation. PMID- 12721992 TI - Pelvic adhesions. PMID- 12721991 TI - Laparoscopic ovarian cystectomy using double balloon catheter: a new technical application. PMID- 12721993 TI - Evaluation of bipolar technology for laparoscopic supracervical hysterectomy. PMID- 12721995 TI - Use of the PumpVac Plus Suction Irrigation System at Gynecologic Endoscopy. PMID- 12721994 TI - Ultrasonic surgical aspiration for gynecologic disease. PMID- 12721996 TI - Radiofrequency Thermal Therapy for Benign Prostatic Hyperplasia by Transurethral Needle Ablation of the Prostate (TUNA): An Update. PMID- 12721997 TI - Interstitial laser coagulation of the prostate: minimally invasive therapy for benign prostatic hyperplasia. PMID- 12721998 TI - Expanding the role of technology: stereotactic breast biopsy in the mobile environment. PMID- 12722000 TI - The impact of genetic screening on surgical decision-making in breast cancer. PMID- 12721999 TI - Thoracoscopic parasternal lymph node dissection for the staging of breast cancer. PMID- 12722001 TI - Advances in the treatment of metastatic colorectal cancer. PMID- 12722002 TI - A modular system for cemented and cementless implantation of femoral stem prostheses. PMID- 12722003 TI - The use of hydroxyapatite-coated implants in revision surgery for acetubular component loosening. PMID- 12722004 TI - Hydroxyapatite Coated (HAC) Femoral Component. PMID- 12722006 TI - Use of external minifixators in hand pathology. PMID- 12722005 TI - Factors affecting stability of press fit acetubular cups. PMID- 12722007 TI - Results of prescription use of pulse ultrasound therapy in fracture management. PMID- 12722008 TI - Absorbable polyglycolide and polylactide devices for fracture fixation. PMID- 12722009 TI - Techniques of posterior cervical spine fusion. PMID- 12722010 TI - Hydroxyapatite cement, a smart biomaterial for craniofacial skeletal tissue engineering. PMID- 12722012 TI - The use of radiosurgery in plastic surgery and dermatology. PMID- 12722011 TI - Alternative Total TMJ Arthroplasty: Metal-on-Metal for Longevity in Implant Survivorship and Patient Satisfaction. PMID- 12722013 TI - Vertical mammaplasty: technique and complications. PMID- 12722014 TI - Image-guided neurosurgery. PMID- 12722015 TI - The use of oxidized regenerated cellulose in neurosurgical procedures. PMID- 12722017 TI - Pemetrexed (Alimta): a promising new agent for the treatment of breast cancer. AB - Pemetrexed (Alimta; Eli Lilly and Company, Indianapolis, IN) is a new antifolate drug with activity at multiple points in folate metabolism, including thymidylate synthase, dihydrofolate reductase, and glycinamide ribonucleotide formyltransferase. In this way it inhibits the de novo synthesis of both purines and pyrimidines. In phase II studies, pemetrexed has shown good clinical activity as single-agent therapy in patients with advanced breast cancer with or without prior treatment, including patients with prior anthracycline and taxane treatment. Toxicities are primarily neutropenia, mucositis, and skin rash. Hematologic toxicities are markedly reduced by supplementation with vitamin B(12) and folate. Skin rash is ameliorated with prophylactic corticosteroid treatment. Pemetrexed is a promising agent in breast cancer and warrants further investigation in this setting. PMID- 12722018 TI - Monotherapy options in the management of metastatic breast cancer. AB - Anthracyclines and taxanes are the most active agents in the first-line treatment of metastatic breast cancer. Among the options following clinical resistance to anthracycline and/or taxane treatment, the most widely used cytotoxic agents are the vinca alkaloid vinorelbine, the fluoropyrimidine capecitabine, and the nucleoside analogue gemcitabine. No direct comparisons of these agents have been reported in single-agent treatment or in single-agent versus combination treatment. Review of data from phase II trials in metastatic breast cancer indicates that the agents produce comparable response rates and are well tolerated. Review of the relatively smaller amount of data on effects in exclusively anthracycline- or anthracycline-taxane-refractory patient groups yields a similar conclusion. Given the absence of definitive comparisons of these agents, and any marked survival difference on informal comparison, treatment decisions should be aimed at maximizing patient benefit by considering the overall benefit of treatment in relation to the potential harm. A similar approach should be applied when considering use of combinations of these agents versus single-agent treatment. PMID- 12722019 TI - Gemcitabine and anthracyclines in breast cancer. AB - Anthracyclines are among the most active drugs in breast cancer. Gemcitabine is a novel agent that has also shown good antitumor activity in advanced breast cancer. This compound has a favorable toxicity profile, no apparent multidrug resistance, and retains its activity in anthracycline-pretreated patients. Therefore, it was logical to combine gemcitabine with anthracyclines. The combinations of gemcitabine and anthracyclines (doxorubicin, liposomal doxorubicin, epirubicin) have been tested in a few phase I and II studies. Generally, these combinations have been shown to be feasible and well tolerated. The pharmacokinetics of gemcitabine and anthracyclines was not affected by their combined use. Myelosuppression was the most common adverse event; neutropenia was easily reversible. Data from phase II clinical studies suggest that gemcitabine/anthracycline combinations are relatively effective in first- or second-line therapy for women with locally advanced and metastatic breast cancer. Future randomized clinical trials are warranted to further elucidate the role of these regimens in breast cancer care. PMID- 12722020 TI - Gemcitabine combined with docetaxel in metastatic breast cancer. AB - The combination of gemcitabine and docetaxel exhibits good activity in the treatment of advanced breast cancer. Phase II trial response rates using 21- or 28-day schedules have ranged from 36% to 79% in patient populations with varying degrees of pretreatment. In studies of extensively pretreated anthracycline resistant patients, response rates have ranged from 36% to 54%, with median times to disease progression of 7 to 8 months and median survival of 12.7 to 14 months. In less treatment-experienced patients, the combination produced an overall response rate of 79%, median time to progression of 7.6 months, and median survival of 24.5 months. Toxicity has primarily consisted of neutropenia. The combination warrants additional investigation as front-line chemotherapy and in patients with anthracycline-resistant and taxane-resistant advanced disease. PMID- 12722021 TI - Gemcitabine combined with paclitaxel or paclitaxel/trastuzumab in metastatic breast cancer. AB - The combination of gemcitabine and paclitaxel in metastatic breast cancer is attractive because of the good single-agent activity of each, the different mechanisms of antitumor effect, and the, generally, nonoverlapping toxicity profiles of the agents. Phase II trials of the combination indicate response rates of 45% to 55% and median survival of 12 months in extensively pretreated patients, including those with anthracycline-resistant disease, and a response rate of 69% in patients with newly recurrent metastatic breast cancer. The combination was well tolerated, with severe hematologic toxicity being infrequent. The combination of gemcitabine, paclitaxel, and trastuzumab in patients with HER2-positive metastatic breast cancer with no prior chemotherapy in the metastatic setting produced a response rate of 71% and median response duration of 11 months. The combination was safe and well tolerated, with no unexpected toxicities being observed. Both combinations warrant additional study in metastatic breast cancer. PMID- 12722023 TI - Making treatment choices: a physician's discussion of patients' needs. AB - Medical treatment decisions in early as well as advanced breast cancer follow established guidelines clearly defined by consensus statements, meta-analyses, and evidence-based insights into the biology of the disease, and treatment efficacy. Nevertheless, to treat patients "holistically" while remaining mindful of individual characteristics, physicians must consider patients' concerns regarding the course of their disease, their distress, and, in advanced disease, their knowledge of a limited life expectancy when making treatment choices. Such considerations will contribute to a more satisfactory patient-physician relationship and superior quality of life. PMID- 12722022 TI - Gemcitabine and trastuzumab in metastatic breast cancer. AB - The background, rationale, and preliminary results of a phase II trial of gemcitabine plus trastuzumab in heavily pretreated metastatic breast cancer patients are presented. Gemcitabine and trastuzumab both exhibit single-agent activity in previously treated metastatic breast cancer. Preclinical studies of gemcitabine and trastuzumab in combination showed additive or synergistic antitumor effects in human breast cancer cell lines that overexpress HER2. A multicenter phase II trial was thus conducted to define the safety and efficacy of gemcitabine/trastuzumab in patients previously treated for metastatic breast cancer. Women with measurable metastatic breast cancer whose primary or secondary tumor overexpressed HER2 were eligible for inclusion. Gemcitabine 1,200 mg/m(2) was administered on days 1 and 8 of a 21-day cycle. Trastuzumab was administered at a loading dose of 4 mg/kg initially, then at 2 mg/kg weekly thereafter. Preliminary analysis of 38 evaluable patients showed that gemcitabine/trastuzumab was well tolerated and had significant antitumor activity in this patient population. The study is now concluded, and final analysis of the data is nearing completion. Publication of the results is anticipated in 2003. PMID- 12722024 TI - Antisense therapy directed to protein kinase C-alpha (Affinitak, LY900003/ISIS 3521): potential role in breast cancer. AB - As our understanding of the biology of cancer increases, the attempts to target specific molecules associated with the promotion of cancer are accelerating. One of the targets currently being studied as an important tumor-promoting factor is protein kinase C-alpha (PKC-alpha). To specifically block PKC-alpha, antisense oligonucleotides have been developed, including LY900003 (Affinitak, ISIS-3521; Eli Lilly and Company, Indianapolis, IN), which is currently in clinical development. Although its single-agent activity in breast cancer is modest, its potential role may be in concert with traditional chemotherapy. This is a review of the pharmacology and current status of the clinical development of LY900003 and its potential role in treating patients with breast cancer. PMID- 12722025 TI - Chronic kidney disease: issues and establishing programs and clinics for improved patient outcomes. AB - The spectrum of chronic kidney disease (CKD) extends from the point at which there is slight kidney damage, but normal function, to the point at which patients require either a renal transplant or renal replacement therapy to survive. Epidemiological studies suggest there are approximately 20,000,000 patients with various stages of CKD. These patients have many comorbidities, including cardiovascular disease, hypertension, diabetes, anemia, nutritional and metabolic derangements, and fluid overload. Unfortunately, evidence shows that current CKD care in the United States is suboptimal, and late referral to a nephrologist is often the rule and not the exception. Roles of primary care physicians (PCPs) and nephrologists in the care of patients with CKD remain undefined. Several studies have suggested that care provided by multidisciplinary nephrology teams can improve patient outcomes. Currently, there are published evidence-based clinical practice guidelines for anemia management, nutritional therapy, and vascular access placement, with other CKD guidelines under development. The intent of this review includes providing compelling evidence for earlier screening, identification, and management of patients with CKD; showing that current CKD care is suboptimal; encouraging the development of multidisciplinary teams that provide collaborative care to patients with CKD, suggesting roles for PCPs and nephrologists in the care of these patients; describing CKD initiatives from national organizations; and providing a comprehensive checklist that can guide the development of CKD clinics and programs. PMID- 12722026 TI - Obesity and survival on dialysis. AB - Several factors associated with greater cardiovascular mortality in the general population may show a paradoxical relationship in patients on dialysis therapy. This dialysis-risk paradox has been reported for high blood pressure, serum lipid levels, and body mass, but the finding is more consistent and persuasive for obesity. This article examines the literature on the association between body mass and dialysis survival and considers the possible mechanistic and clinical implications. PMID- 12722027 TI - Proinflammatory genotype of interleukin-1 and interleukin-1 receptor antagonist is associated with ESRD in proteinase 3-ANCA vasculitis patients. AB - BACKGROUND: Small-vessel vasculitides are associated with antineutrophil cytoplasmic antibodies (ANCAs). Cytoplasmic ANCAs are targeted mainly against proteinase 3 (PR3), whereas myeloperoxidase (MPO) is the major antigen of perinuclear ANCAs. These relapsing vasculitides show heterogeneous clinical pictures, and disease severity may vary broadly from mild local organ manifestation to acute organ failure (eg, renal failure). We tested whether two cytokine polymorphisms in the interleukin-1beta (IL-1beta) and IL-1 receptor antagonist (IL-1ra) genes, known to determine cytokine secretion, are associated with clinical manifestations and outcome of ANCA-associated vasculitides. METHODS: Polymerase chain reaction and restriction fragment length polymorphism analyses were performed to determine polymorphisms in the IL-1beta and IL-1ra genes in 79 patients with PR3-ANCA, 30 patients with MPO-ANCA vasculitis, and 196 healthy controls. RESULTS: The frequency of the so-called proinflammatory genotype, characterized by high secretion of IL-1beta and low secretion of its antagonist IL-1ra, was increased significantly in patients with PR3-ANCA with end stage renal disease. CONCLUSION: Patients with a renal manifestation of PR3-ANCA vasculitis have an increased risk for developing end-stage renal disease when carrying the proinflammatory IL-1beta/IL-1ra genotype. Anti-inflammatory therapy specifically antagonizing the proinflammatory effect of IL-1beta may be a promising treatment for patients with Wegener's granulomatosis with renal manifestations. PMID- 12722028 TI - ACE gene polymorphism and progression of diabetic nephropathy in Korean type 2 diabetic patients: effect of ACE gene DD on the progression of diabetic nephropathy. AB - BACKGROUND: Pathophysiological causes of the development and progression of diabetic nephropathy are not well known, but the angiotensin-converting enzyme (ACE) gene polymorphism has been proposed to be involved in its development and progression. METHODS: The impact of insertion/deletion (I/D) genotypes on the progression of diabetic nephropathy in 239 Korean patients with type 2 diabetes (99 patients with stable renal function, group 1; 140 patients with declining renal function, group 2) was investigated by retrospective review of clinical data. RESULTS: The frequency of the DD genotype was significantly greater in group 2 compared with group 1 (30.7% versus 9.1%; P < 0.05). There were no significant differences in age, blood pressure, hemoglobin A(1c) levels, or lipid profiles among ACE genotype groups. However, the prevalence of retinopathy was significantly greater in patients with the DD genotype (DD, ID, and II, 90.4%, 71.2%, and 70.6%, respectively; P < 0.05). Patients with the DD genotype reached the end point (serum creatinine > 2.0 mg/dL [176.8 micromol/L]) faster than those with the other genotypes (DD, 11.38 +/- 4.08 years; ID, 13.85 +/- 4.04 years; II, 14.04 +/- 4.06 years, respectively; P < 0.05) and took significantly less time to reach dialysis therapy (DD, 13.10 +/- 4.45 years; ID, 16.21 +/- 4.74 years; II, 15.13 +/- 4.09 years, respectively; P < 0.05). In multiple logistic regression analysis, systolic blood pressure and DD genotype showed significant correlations with the progression of diabetic nephropathy. In patients with the DD genotype, the odds ratio was 3.881 (95% confidence interval, 1.564 approximately 9.628; P = 0.003) compared with those with the II genotype. CONCLUSION: It is suggested that the ACE gene DD genotype might be a significant risk factor for the progression of diabetic nephropathy. PMID- 12722029 TI - Costimulatory molecules CD80 and CD86 in human crescentic glomerulonephritis. AB - BACKGROUND: CD80 and CD86, cell-surface molecules found only on antigen presenting cells (APCs), are required for activation of CD4-postitve (CD4+) T cells by interaction with CD28/cytotoxic T-lymphocyte-associated antigen 4 on T cells. The roles of these molecules in human glomerulonephritis (GN) presently are unknown. METHODS: Twelve cases of crescentic GN, thought to be a T helper cell-directed delayed-type hypersensitivity-like injury, and 10 controls with non immunoglobulin A proliferative GN were used. Expression of CD80, CD86, CD4, CD14, CD68, HLA-DR, and intercellular adhesion molecule-1 was investigated in renal tissues using monoclonal antibodies and compared with clinical data at the time of renal biopsy. RESULTS: CD80+ and CD86+ cells were observed significantly more in crescentic GN than in controls. CD86 was expressed in the glomerulus and interstitium, especially in the crescent, and adhesion to Bowman's capsule and periglomerular areas corresponding to these changes. Tubular epithelial cells showed no CD86 expression, but they expressed CD80, and some of them expressed HLA-DR. CD4, CD14, CD68, and CD86 showed similar distribution patterns. Positive correlations were found between CD86+ cells and CD4+, CD14+, and CD68+ cells. The number of interstitial CD86+ cells correlated with deterioration of renal function. Most CD86+ cells were monocyte/macrophages. CONCLUSION: This study suggests that the costimulatory molecules CD80 and CD86 have different expressions in human crescentic GN, and CD86 is concerned with crescent formation and CD4+ T-cell accumulations. The majority of APCs are macrophages, and tubular cells also can act as APCs. PMID- 12722030 TI - High urine volume and low urine osmolality are risk factors for faster progression of renal disease. AB - BACKGROUND: Increased fluid intake slows renal disease progression in animal models. The relevance of these findings to human renal disease is not clear, although increased fluid intake often is recommended to patients with chronic renal insufficiency. This study tested the hypothesis that urine volume, urine osmolality (Uosm), or both are significantly associated with glomerular filtration rate (GFR) decline in patients with chronic renal insufficiency. METHODS: This is a retrospective analysis of Modification of Diet in Renal Disease (MDRD) study A patients with (N = 139) and without polycystic kidney disease (PKD; N = 442). The key outcome measure was GFR slope in relation to mean 24-hour urine volume and Uosm during follow-up in study A (mean, 2.3 years). RESULTS: The regression of GFR slope on mean follow-up 24-hour urine volume (adjusted for body surface area and MDRD diet and blood pressure group) showed that the greater the urine volume, the faster the GFR decline in patients both with and without PKD. For example, the difference in GFR slope for those with a mean follow-up 24-hour urine volume of 2.4 versus 1.4 L was -1.01 mL/min/y (confidence interval, -0.27 to -1.75) for patients without PKD and -1.20 mL/min/y (confidence interval, -0.06 to -2.34) for those with PKD. A similar but inverse relationship was shown between GFR decline and mean 24-hour Uosm in patients with (P = 0.01) and without PKD (P = 0.001). These associations remained significant after adjustment for 13 relevant baseline and follow-up covariates. CONCLUSION: Sustained high urine volume and low Uosm are independent risk factors for faster GFR decline in patients with chronic renal insufficiency. Thus, high fluid intake does not appear to slow renal disease progression in humans. We suggest that until better evidence becomes available, patients with chronic renal insufficiency should generally let their thirst guide fluid intake. The advice to avoid "pushing fluids" might be particularly important for patients with PKD. PMID- 12722031 TI - Controlled, prospective trial of steroid treatment in IgA nephropathy: a limitation of low-dose prednisolone therapy. AB - BACKGROUND: No accepted therapy has been established for progressive immunoglobulin A (IgA) nephropathy. METHODS: A prospective, randomized, controlled trial of low-dose prednisolone therapy was performed in patients with IgA nephropathy with moderate histological characteristics. Forty-three patients in the steroid group and 47 patients in the control group were included in the study. The initial dose of prednisolone was 20 mg/d, gradually tapered to 5 mg/d during 2 years. RESULTS: Baseline urine protein-creatinine ratio (UP-UCR) was significantly greater in the steroid group than in controls. Follow-up duration was 65 +/- 25 months in the steroid group and 64 +/- 23 months in controls. Changes in UP-UCR from baseline, ie, UP-UCR at last follow-up minus UP-UCR at baseline, were significantly lower in the steroid group than in controls (steroid group, -0.84 +/- 1.78; controls, 0.26 +/- 1.65; P = 0.0034). Kidney survival was similar in both groups. Patients were divided into two subgroups according to clinical course. There were 28 improved patients and 15 unimproved patients in the steroid group and 27 improved patients and 20 unimproved patients in the control group. In the steroid group, UP-UCR was significantly greater in the unimproved than improved subgroup (3.1 +/- 2.6 versus 1.8 +/- 1.5). CONCLUSION: These data suggest that our protocol had an antiproteinuric effect, but could not improve kidney survival. Because the effect of steroid therapy to prevent the progression of IgA nephropathy is believed to be linked closely to reduction in urinary protein, an insufficient dose of prednisolone in our protocol may be the reason for the discrepancy between the effect on proteinuria and kidney survival. PMID- 12722032 TI - Continuous renal replacement therapy in children up to 10 kg. AB - BACKGROUND: There is growing use of continuous renal replacement therapy (CRRT) for pediatric patients, but no large studies reporting CRRT use and outcome in young children. We describe a cohort of patients weighing 10 kg or less who underwent CRRT at five US children's hospitals between 1993 and 2001. METHODS: We reviewed records of 85 patients weighing 10 kg or less who underwent at least 24 hours of CRRT. We evaluated weight, diagnosis, pressor number, CRRT characteristics, days on CRRT, and outcome (survival to leave intensive care unit versus death). RESULTS: Patients weighed 1.5 to 10 kg (mean, 5.3 +/- 2.8 kg; 16 patients < or = 3 kg). Sixty-nine percent of patients were being administered pressors at the time of CRRT initiation, 88% of patients were administered heparin, and the others were administered citrate or no anticoagulation. Mean blood flow was 48 +/- 24 mL/min (range, 15 to 106 mL/min) or 9.5 +/- 4.2 mL/min/kg. Six hundred fifty-five patient-days of therapy were studied (mean, 7.6 +/- 8.6 d/patient; range, 1 to 46 d/patient). Thirty-two patients (38%) survived; 4 of 16 patients (25%) weighing 3 kg or less survived. The smallest survivor weighed 2.3 kg. Overall, survivors and nonsurvivors showed no significant difference in weight, days on CRRT, or pressor number. However, for patients weighing more than 3 kg, 28 of 69 patients (41%) survived, and mean pressor number was lower for survivors versus nonsurvivors (0.96 +/- 1.1 versus 1.6 +/- 1.0 pressors; P < 0.03). CONCLUSION: CRRT is feasible and useful in children weighing 10 kg or less. Hemodynamic instability requiring pressor support neither precludes successful CRRT nor adversely affects survival. After CRRT, the survival rate in children who weigh 3 to 10 kg is similar to that in older children and adolescents. PMID- 12722033 TI - Relationships between renovascular disease, blood pressure, and renal function in the elderly: a population-based study. AB - BACKGROUND: The purpose of this study is to examine the associations between renovascular disease (RVD) and cross-sectional measures of blood pressure and renal function among participants in the Cardiovascular Health Study (CHS). METHODS: The CHS is a prospective cohort study of cardiovascular disease among elderly Americans. As part of an ancillary study, participants in the Forsyth County, NC, cohort of the CHS were invited to undergo renal duplex sonography (RDS) to define the presence or absence of RVD (defined as any focal peak systolic velocity > or = 1.8 milliseconds or the absence of a Doppler shifted signal from an imaged artery). Demographic, risk factor, blood pressure, and serum creatinine data were obtained at the time of RDS and from the annual CHS examination. RESULTS: Eight hundred thirty-four CHS participants (including 525 women [63%], 309 men [37%], 194 African Americans [23%], and 635 Caucasians [76%]) with a mean age of 77.2 +/- 4.9 years underwent successful RDS. RVD was present in 57 participants (6.8%). When examined according to the presence or absence of RVD, significant univariate differences were observed in the prevalence of clinical hypertension (72% versus 50%; P = 0.001), systolic blood pressure (145 versus 136 mm Hg; P = 0.001), and renal insufficiency (16% versus 8%; P = 0.041). Multivariate analyses showed significant and independent associations for the presence of RVD with increasing systolic blood pressure (P = 0.034), clinical hypertension (odds ratio, 2.68; 95% confidence interval, 1.44 to 4.99; P = 0.002), increasing serum creatinine level, and renal insufficiency (odds ratio, 2.21; 95% confidence interval, 1.02 to 4.79; P = 0.043). A significant interaction was observed between the presence of RVD and increasing systolic blood pressure in association with increasing serum creatinine levels (P = 0.041). CONCLUSION: These results suggest important population-based associations between RVD and cross-sectional measures of blood pressure and renal function. Furthermore, the observed relationship between RVD and increasing serum creatinine level was influenced strongly by increasing blood pressure. PMID- 12722034 TI - Useful biochemical markers for diagnosing renal osteodystrophy in predialysis end stage renal failure patients. AB - BACKGROUND: Various biochemical markers have been evaluated in dialysis patients for the diagnosis of renal osteodystrophy (ROD). However, their value in predialysis patients with end-stage renal failure (ESRF) is not yet clear. METHODS: Bone histomorphometric evaluation was performed and biochemical markers of bone turnover were determined in serum of an unselected predialysis ESRF population (N = 84). RESULTS: Significant (P < 0.005) differences between the five groups with ROD (ie, normal bone [N = 32], adynamic bone [ABD; N = 19], hyperparathyroidism [N = 8], osteomalacia [OM; N = 10], and mixed lesion [N = 15]) were noted for intact parathyroid hormone, total (TAP) and bone alkaline phosphatase (BAP), osteocalcin (OC), and serum calcium levels. Serum creatinine and (deoxy)pyridinoline levels did not differ between groups. For the diagnosis of ABD, an OC level of 41 microg/L or less (< or =7.0 nmol/L) had a sensitivity of 83% and specificity of 67%. The positive predictive value (PPV) for the population under study was 47%. The combination of an OC level of 41 ng/L or less (< or =7.0 nmol/L) with a BAP level of 23 U/L or less increased the sensitivity, specificity, and PPV to 72%, 89%, and 77%, respectively. ABD and normal bone taken as one group could be detected best by a BAP level of 25 U/L or less and TAP level of 84 U/L or less, showing sensitivities of 72% and 88% and specificities of 76% and 60%, corresponding with PPVs of 89% and 85%, respectively. In the absence of aluminum or strontium exposure, serum calcium level was found to be a useful index for the diagnosis of OM. CONCLUSION: OC, TAP, BAP, and serum calcium levels are useful in the diagnosis of ABD, normal bone, and OM in predialysis patients with ESRF. PMID- 12722035 TI - A comparison of dual dialyzers in parallel and series to improve urea clearance in large hemodialysis patients. AB - BACKGROUND: Dialysis adequacy targets frequently are difficult to achieve in large hemodialysis patients. Dual dialyzers can be used to improve clearance. It is unknown whether series or parallel configurations are superior. METHODS: Eighteen large hemodialysis patients (mean weight, 92.4 kg) were enrolled in a randomized, crossover trial to directly compare dual dialyzers in parallel and series configurations. Treatment times, blood flow rates, and dialysate flow rates were kept constant. RESULTS: Compared with a single dialyzer, parallel dual dialyzers increased the single-pool Kt/V (spKt/V) from 1.25 +/- 0.22 to 1.43 +/- 0.29 (P < 0.003). Series dual dialyzers improved urea clearance measured by spKt/V (spKt/V(urea)) to 1.46 +/- 0.26 (P < 0.0003 compared with a single dialyzer). Kt/V and urea reduction ratio of dual dialyzers in parallel were not significantly different from those of dual dialyzers in series. Half the subjects failed to meet the National Kidney Foundation-Kidney Disease Outcomes Quality Initiative recommended adequacy target for spKt/V(urea) of 1.2 or less using a single dialyzer. With the use of dual dialyzers, 83% of subjects achieved this adequacy target. Serum levels of a middle molecule, beta2-microglobulin, were reduced 34% after 2 months of dual-dialyzer therapy. Cost analysis estimates annual net savings of 1,260 dollars with dual-dialyzer therapy, primarily from projected savings in inpatient expenses. CONCLUSION: In large hemodialysis patients, our study shows that dual dialyzers in parallel and series are equally effective at improving urea clearance without prolonging dialysis treatment times. PMID- 12722036 TI - Correlates of subclinical left ventricular dysfunction in ESRD. AB - BACKGROUND: Abnormalities of the left ventricle are common in patients with end stage renal disease (ESRD) both before and after the start of renal replacement therapy. The purpose of this study is to identify possible causes of subclinical left ventricular (LV) dysfunction in patients with ESRD. In particular, we sought to determine whether the presence of ESRD was itself associated with dysfunction independent of LV hypertrophy and coronary artery disease. METHODS: Assessment of cardiovascular risk factors and dialysis adequacy was completed in 145 unselected patients with ESRD who were recruited from the renal dialysis unit and compared with age- and sex-matched controls. Among the 68 patients with ESRD who had undergone a dobutamine stress echocardiogram with normal findings, regional cardiac function was quantified by myocardial Doppler velocity, LV volumes and mass were measured using three-dimensional echocardiography, and vascular function was assessed using brachial artery reactivity (BAR). RESULTS: LV diastolic velocity was impaired in patients with ESRD, but there was no significant difference in systolic velocity compared with control patients of similar age. Age, diabetes mellitus, hypertension, and LV mass were independent predictors of diastolic velocity (model R2 = 0.45; P < 0.001), whereas age and risk factor number were predictors of systolic velocity (model R2 = 0.19; P = 0.002). Increasing risk factor number had no significant relationship with LV mass or volume. There was no detected association between BAR and incremental risk factors (P = 0.51). CONCLUSION: Subclinical LV dysfunction occurs in patients with ESRD, but is evidenced as abnormal myocardial diastolic, rather than systolic, function. Correlates of abnormal function are age, diabetes mellitus, hypertension, and LV mass, rather than ESRD alone, dialysis adequacy, or abnormal endothelial function. PMID- 12722037 TI - Estimation of delivered dialysis dose by on-line monitoring of the ultraviolet absorbance in the spent dialysate. AB - BACKGROUND: Several methods are available to determine Kt/V, from predialysis and postdialysis blood samples to using on-line dialysate urea monitors or to ionic dialysance using a conductivity method. The aim of this study is to compare Kt/V calculated from the slope of the logarithmic on-line ultraviolet (UV) absorbance measurements, blood urea Kt/V, dialysate urea Kt/V, and Kt/V from the Urea Monitor 1000 (UM; Baxter Healthcare Corp, Deerfield, IL). METHODS: Thirteen uremic patients on chronic thrice-weekly hemodialysis therapy were included in the study. The method uses absorption of UV radiation by means of a spectrophotometric set-up. Measurements were performed on-line with the spectrophotometer connected to the fluid outlet of the dialysis machine; all spent dialysate passed through a specially designed cuvette for optical single wavelength measurements. UV absorbance measurements were compared with those calculated using blood urea and dialysate urea, and, in a subset of treatments, the UM. RESULTS: Equilibrated Kt/V (eKt/V) obtained with UV absorbance (eKt/Va) was 1.19 +/- 0.23; blood urea (eKt/Vb), 1.30 +/- 0.20, and dialysate urea (eKt/Vd), 1.26 +/- 0.21, and Kt/V in a subset measured by the UM (UM Kt/V) was 1.24 +/- 0.18. The difference between eKt/Vb and eKt/Va was 0.10 +/- 0.11, showing a variation similar to the difference between eKt/Vb and eKt/Vd (0.03 +/- 0.10) and in a subset between eKt/Vb and UM Kt/V (-0.02 +/- 0.11). CONCLUSION: The study suggests that urea Kt/V can be estimated by on-line measurement of UV absorption in the spent dialysate. PMID- 12722038 TI - Association of depression with malnutrition in chronic hemodialysis patients. AB - BACKGROUND: Depression is the most common psychological complication and may increase mortality in chronic hemodialysis patients. Because depression could be associated with poor oral intake and activation of proinflammatory cytokines that could further increase mortality by malnutrition, we investigated the relation between depression and nutritional status in chronic hemodialysis patients. METHODS: Sixty-two Korean patients completed the Beck Depression Inventory (BDI) questionnaire, and the diagnosis of depression was confirmed by Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) criteria for major depressive disorder. Nutritional status was evaluated using serum albumin level, normalized protein catabolic rate, subjective global assessment (SGA), and anthropometric measurement. RESULTS: Mean BDI score was 22.7 +/- 11.4, and 35 patients (56.5%) had a BDI score greater than 21, which is the suggested cutoff score for the diagnosis of depression for the Korean population. Of 40 patients who had a score higher than 18 on the BDI, 34 patients met DSM-IV criteria for major depressive disorder. BDI score correlated negatively with a variety of nutritional parameters: serum albumin level (r = -0.47; P < 0.001), normalized protein catabolic rate (r = -0.32; P < 0.05), SGA (r = -0.47; P < 0.01), triceps skinfold thickness (r = -0.40; P < 0.05), midarm muscle circumference (r = -0.57; P < 0.01), and body mass index (r = -0.28; P < 0.05). Multiple regression analysis also identified BDI score as an independent determinant for all kinds of nutritional parameters. CONCLUSION: In patients on chronic hemodialysis therapy, depression is related closely to nutritional status and could be an independent risk factor for malnutrition. PMID- 12722040 TI - Correlation between histomorphometric parameters of bone resorption and serum type 5b tartrate-resistant acid phosphatase in uremic patients on maintenance hemodialysis. AB - BACKGROUND: Serum tartrate-resistant acid phosphatase 5b (TRACP) is a new marker of potential clinical use to monitor osteoclastic activity and bone resorption rate. The relationship between histomorphometric parameters of bone resorption and serum TRACP was evaluated in 14 chronically dialyzed patients and 6 healthy control subjects. METHODS: All patients underwent bone biopsies and serum biochemical testing for TRACP, intact parathyroid hormone (iPTH), pyridinoline cross-linked telopeptide domain of type I collagen (ICTP), total calcium, phosphorus, and albumin, which were measured at the time of biopsy. RESULTS: Bone histological examination showed predominant hyperparathyroid bone disease (HPT) in 6 patients, mixed uremic osteodystrophy in 3 patients, low-turnover osteomalacia in 1 patient, and adynamic bone disease in 4 patients. Mean TRACP activity was 3.25 +/- 0.59 U/L in control subjects. Median TRACP activity was significantly greater in patients with HPT (11.97 +/- 8.92 U/L) than those with other types of renal osteodystrophy (ROD; 2.17 +/- 0.61 U/L). Serum iPTH levels were greatest in all patients with HPT, but also were significantly elevated in 7 of 8 patients with other types of ROD. Serum ICTP levels also were significantly elevated in all patients with HPT and 6 of 8 patients with other types of ROD. Serum TRACP levels correlated more strongly with histological parameters of osteoclasts than those of erosion. Also, correlations between TRACP and histological parameters of osteoclasts were stronger than those of iPTH and ICTP levels. CONCLUSION: These early results suggest that serum TRACP levels correlate well with histological indices of osteoclasts and may serve as a specific marker for osteoclastic activity in patients with renal bone disease. PMID- 12722039 TI - Soluble Fas: a novel predictor of atherosclerosis in dialysis patients. AB - BACKGROUND: Cardiovascular disease (CVD) is the leading cause of death in patients with end-stage renal disease (ESRD). Disregulation of apoptosis within the vessel wall and upregulation of the Fas/Fas-ligand (Fas-L) system contribute to the development of atherosclerosis. Cross-sectional studies have suggested that elevated plasma levels of the soluble form of Fas (sFas) are associated with CVD. However, the role of sFas and sFas-L in predicting future cardiovascular events has yet to be defined. METHODS: We evaluated the role of plasma sFas and sFas-L levels as predictors of CVD in a prospective cohort of 107 chronic hemodialysis patients. RESULTS: During the study period (27 months), 53 patients (49.5%) presented with at least one cardiovascular end point. On univariate analysis, baseline sFas levels were significantly associated with the occurrence of cardiovascular end points, whereas sFas-L levels were not. Using Cox proportional hazards, increased sFas levels were associated with a significantly greater risk for cardiovascular end points (P = 0.03). This effect was independent of baseline CVD history, classic risk factors for atherosclerosis (diabetes, hypercholesterolemia, hypertension, and smoking), and markers of inflammation (C-reactive protein [CRP], soluble intercellular adhesion molecule 1). Increased CRP levels also were associated with cardiovascular end points (P = 0.04). In addition, increased cardiovascular mortality was found in patients in the highest sFas tertile compared with those in the lowest tertile (27.8% versus 8.6%; P = 0.04). CONCLUSION: Increased plasma sFas levels are predictive of future CVD. These results suggest that sFas is a novel and independent predictor of active atherosclerotic disease in patients with ESRD. PMID- 12722041 TI - Is dialysis modality a factor in survival of patients with ESRD and HIV associated nephropathy? AB - BACKGROUND: Human immunodeficiency virus (HIV)-associated nephropathy (HIVAN), characterized by a fulminant form of focal segmental glomerulosclerosis, has become the third leading cause of end-stage renal disease (ESRD) in young African Americans. There is a theoretical possibility that hemodialysis (HD) therapy in these patients may enhance HIV replication through the activation of white blood cells and release of such cytokines as tumor necrosis factor-alpha, interleukin 1, and interleukin-6, which have been found to increase HIV replication in vitro. We therefore determined whether dialysis modality is a factor in the survival of patients with HIVAN and ESRD. METHODS: Information regarding dialysis modality was available for 6,053 of 6,166 patients with ESRD and HIVAN who started dialysis therapy in the United States from December 1995 to December 1999 by using the US Renal Data System database. RESULTS: Eighty-nine percent were black. Eighty-eight percent underwent HD, and 12%, peritoneal dialysis (PD). On Cox proportional hazard analysis, after adjusting for demographic variables and year of dialysis therapy initiation, there was no difference in survival between the different modalities (PD versus HD: hazard ratio, 1.01; 95% confidence interval, 0.91 to 1.13). In addition, on censoring patients at the time of first dialysis modality switch, no difference in survival was found between PD and HD. CONCLUSION: We conclude that patients with HIVAN and ESRD should be given an option to choose dialysis modality because it is not a factor in predicting survival. PMID- 12722042 TI - Structural and functional correlations in stable renal allografts. AB - BACKGROUND: Renal functional reserve (RFR) has been proposed as a surrogate marker of renal mass, but its significance in well-functioning renal transplants is controversial. Thus, we used early protocol biopsies to analyze structural and functional correlations in stable grafts. METHODS: We studied 32 cyclosporine (CsA)-treated stable cadaveric transplants at 5 months. Biopsies were evaluated according to Banff criteria and histomorphometry. Inulin and p-aminohippurate clearances were used to calculate glomerular filtration rate (GFR) and effective renal plasma flow (ERPF). RFR after an amino acid infusion (RFR-AA) and after a combined amino acid and dopamine infusion (RFR-AA-DOPA) was evaluated. RESULTS: Baseline GFR was 54 +/- 16 mL/min/1.73 m2, and ERPF was 219 +/- 55 mL/min/1.73 m2. RFR-AA was 9% +/- 13%, and RFR-AA-DOPA was 22% +/- 20%. RFR-AA correlated with CsA dose (R = 0.39; P = 0.02), whereas RFR-AA-DOPA correlated with CsA dose (R = 0.36; P = 0.04) and CsA levels (R = 0.40; P = 0.02). The only histological parameter associated with RFR was the presence of arteriolar hyalinosis (AH). Patients showing an AH score of 1 or greater (n = 7) had lower RFR-AA (0% +/- 9% versus 11% +/- 13%; P = 0.02) and lower RFR-AA-DOPA (9% +/- 17% versus 26% +/- 19%; P = 0.03). Multivariate analysis showed that an AH score of 1 or greater, but not CsA dose or levels, was associated with RFR-AA (R = 0.42; P = 0.01). RFR AA-DOPA was associated with hyaline arteriolar damage (R = 0.43; P = 0.01), as well as CsA levels (R = 0.54; P = 0.006). CONCLUSION: The presence of AH is the only histological parameter associated with impaired RFR in well-functioning grafts. PMID- 12722043 TI - Urinary actin, interleukin-6, and interleukin-8 may predict sustained ARF after ischemic injury in renal allografts. AB - BACKGROUND: Cellular damage and inflammation after ischemia contribute to sustained acute renal failure (ARF). METHODS: To quantify cellular damage and inflammation in postischemic ARF and identify markers of renal functional outcome, urine specimens from 40 renal allograft recipients, including 30 cadaveric (9 "sustained ARF" and 21 "recovery" subjects) and 10 living donor allografts ("LD"), were analyzed for actin, gamma-glutamyl transpeptidase (GGTP), lactate dehydrogenase (LDH), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) and interleukin-8 (IL-8) during the first posttransplant week. RESULTS: On day 0, urinary actin, GGTP, IL-6, and IL-8 were elevated in recipients destined to have sustained ARF compared with those destined to recover. Median values per gram of urine creatinine in the sustained ARF, recovery, and LD groups were 263.9, 0.0, and 0.0 microg for actin; 5000.0, 892.9, and 5555.6 U for GGTP; 193.1, 27.2, and 10.5 ng for IL-6; and 382.0, 17.8, and 18.5 ng for IL-8, respectively. In contrast, urinary LDH and TNF-alpha increased in recipients with recovering function compared with those who had sustained ARF. The corresponding median values were 36.7 and 16.3 U (recovery versus sustained ARF) for LDH, and 18.4 and 7.6 ng (LD versus sustained ARF) for TNF-alpha. Computational analyses using the Receiver Operating Characteristic Curve found that elevated urinary actin, IL-6, and IL-8 on day 0 were strong predictors of sustained ARF, where the calculated areas under the curve were 0.75, 0.91, and 0.82, respectively. CONCLUSION: Increased urinary actin, IL-6, and IL-8 may be useful markers for the prediction of sustained ARF after ischemia. PMID- 12722044 TI - Effect of a 3-year therapy with the 3-hydroxy-3-methylglutaryl coenzyme a reductase-inhibitor fluvastatin on endothelial function and distensibility of large arteries in hypercholesterolemic renal transplant recipient. AB - BACKGROUND: In patients after renal transplantation functional arterial vessel wall properties are impaired. Whether 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase inhibitors have a sustained effect on endothelial function and arterial distensibility in patients after renal transplantation is not clear. The authors studied the effects of a long-term therapy with fluvastatin on large artery distensibility and flow-mediated vasodilation (FMD) in hypercholesterolemic patients after renal transplantation in a prospective, blinded, and randomized trial. METHODS: Twenty-six patients who had undergone renal transplantation were assigned randomly to either fluvastatin, 40 mg/d (n = 13) or placebo (n = 13) and underwent follow-up for 3 years. At baseline and after 6, 12, and 36 months of treatment, carotid and brachial artery distensibility, endothelium-dependent FMD, and nitroglycerine-induced vasodilation (NMD) of the brachial artery were measured by a echo-tracking device. RESULTS: A significant decrease in total and low-density cholesterol was observed after 6, 12, and 36 months in patients treated with fluvastatin but not in the placebo group. FMD increased with fluvastatin from 4.6 +/- 2% to 12.4 +/- 2% after 12 months; this improvement was sustained with 13.4 +/- 3% after 36 months (P < 0.05). However, placebo did not alter FMD (P < 0.001 for trend difference between groups by analysis of covariance). Endothelium-independent NMD was similar in both groups at baseline and during therapy. Neither carotid nor brachial artery distensibility coefficients were altered by either treatment. CONCLUSION: HMG-CoA reductase inhibitor therapy over 3 years results in a significant and sustained improvement of endothelial function in hypercholesterolemic patients after renal transplantation. However, this is not accompanied by a beneficial effect on impaired large artery distensibility even after long-term therapy with fluvastatin. PMID- 12722045 TI - IgA antiglomerular basement membrane nephritis associated with Crohn's disease: a case report and review of glomerulonephritis in inflammatory bowel disease. AB - A case of glomerulonephritis in a 35-year-old man with Crohn's disease is described here. The patient presented with severe diarrhea, nephrotic range proteinuria, hematuria, microangiopathic hemolytic anemia, thrombocytopenia, hypocomplementemia, acute renal failure requiring hemodialysis, cryoglobulinemia, and extensive thrombotic gangrene of the distal upper and lower limbs. The patient did not respond to plasmapheresis and steroid therapy and died of upper gastrointestinal bleeding. Renal tissue obtained at autopsy showed IgA-mediated antiglomerular basement membrane crescentic glomerulonephritis. Linear staining of the glomerular basement membrane by non-IgG antibodies is quite unusual with only 11 cases previously reported in the worldwide literature, 8 caused by IgA. Glomerulonephritis is a rarely reported extraintestinal manifestation of inflammatory bowel disease, and there are only 24 previously described cases that are reviewed and summarized in this report. Glomerulonephritis occurred in the setting of active bowel inflammation in all cases, circulating immune complexes were found in nearly half the cases, and serum complements usually were normal. Renal insufficiency and nephrotic range proteinuria were typically present at the time of diagnosis of glomerulonephritis and most often improved in parallel with treatment of the gastrointestinal disorder. The histologic findings were varied and included membranoproliferative glomerulonephritis, mesangioproliferative glomerulonephritis, membranous nephropathy, IgA nephropathy, and IgM nephropathy. Thus, the authors present the first case of glomerulonephritis caused by antiglomerular basement membrane disease in association with inflammatory bowel disease. PMID- 12722046 TI - Alport syndrome-like basement membrane changes in Frasier syndrome: an electron microscopy study. AB - Frasier syndrome (FS) is a rare disease characterized by male pseudohermaphroditism and slowly progressing nephropathy. FS originates from heterozygous mutation in the intron 9 splicing donor site of Wilms' tumor suppressor gene (WT1). Focal segmental glomerular sclerosis is common in FS, but there have not been so many detailed pathologic investigations. The authors examined the kidneys of 3 patients with FS. The results showed that nephropathy started as mesangial proliferative glomerulonephritis, and later a concomitant focal segmental lesion developed. In all cases, electron microscopy results showed widespread thinning, splitting, and lamellation of the glomerular basement membrane, which mimicked hereditary nephritis. Throughout adulthood, WT1 protein expresses on glomerular podocytes. Recent reports described that podocytes expressing WT1 play an important role in maintaining the glomerular basement membrane. Hereditary nephritis-like glomerular basement membrane findings in FS suggest that one of the important functions of podocytes is to form and maintain the glomerular basement membrane. PMID- 12722047 TI - Eosinophil-rich interstitial infiltrate in an allograft biopsy. PMID- 12722048 TI - Intravenous versus subcutaneous EPO: is the cost benefit acceptable? PMID- 12722050 TI - Anemia outcomes in ESRD: individual or facility intervention? PMID- 12722053 TI - Enos polymorphism and renal disease progression in autosomal dominant polycystic kidney disease. PMID- 12722054 TI - Access surgery and the role of nephrologists. PMID- 12722056 TI - Pth 1-84, Pth fragments and bone turnover. PMID- 12722065 TI - Single-molecule-sensitive fluorescent sensors based on photoinduced intramolecular charge transfer. PMID- 12722066 TI - To "dress" the "naked" acetylide with metal complexes: how and why? PMID- 12722062 TI - Quiz page. Systemic amyloidosis. PMID- 12722067 TI - The Pauson-Khand reaction: the catalytic age is here! AB - As a consequence of growing environmental awareness, it is now inappropriate to design a synthetic metal-mediated transformation that involves a noncatalytic use of toxic and expensive transition-metal species. One of the earliest examples of such a metal-mediated transformation is the Pauson-Khand reaction, a [2+2+1] cyclocarbonylation that generates a cyclopentenone. Despite the early descriptions by Pauson and co-workers of catalytic versions of the reaction with octacarbonyldicobalt(0), applications of the Pauson-Khand reaction have to date almost exclusively used approaches that involve stoichiometric quantities of cobalt-carbonyl complexes. In the last decade, and, most markedly, in the last two to three years, however, there have been many exciting and novel developments in the catalytic Pauson-Khand reaction. Furthermore, asymmetric catalysis of the Pauson-Khand reaction has been shown to be a viable process. In view of the impressive developments in Pauson-Khand catalysis in the last two to three years, we present a comprehensive and critical coverage of the catalytic Pauson-Khand reaction that is designed to facilitate its application and to point to exciting future developments. PMID- 12722068 TI - Noncontact liquid-crystal alignment by supramolecular amplification of nanogrooves. PMID- 12722069 TI - Polymerizable bent-core mesogens: switchable precursors to ordered, polar polymer materials. PMID- 12722070 TI - [Cd16In64S134]44-: 31-A tetrahedron with a large cavity. PMID- 12722071 TI - Dendrimer design: how to circumvent the dilemma of a reduction of steps or an increase of function multiplicity? PMID- 12722072 TI - Direct observation of a hydrogen-bonded charge-transfer state of 4 dimethylaminobenzonitrile in methanol by time-resolved IR spectroscopy. PMID- 12722073 TI - Vectorial control of energy-transfer processes in metallocyclodextrin heterometallic assemblies. PMID- 12722074 TI - Synthesis of 2,3,6-trideoxysugar-containing disaccharides by cyclization and glycosidation through the sequential activation of sulfoxide and methylsulfanyl groups in a one-pot procedure. PMID- 12722075 TI - Nanotubes of magnesium borate. PMID- 12722076 TI - Effect of the helix macrodipole on surface-initiated N-carboxyanhydride polymerization on gold. PMID- 12722078 TI - 5,6,7,12-tetrahydrodibenz[c,f][1,5]azabismocines: highly reactive and recoverable organobismuth reagents for cross-coupling reactions with aryl bromides. PMID- 12722077 TI - Detection of paramagnetic pH-dependent inclusion complexes between beta cyclodextrin dimers and nitroxide radicals. PMID- 12722079 TI - Zinc saccharate: a robust, 3D coordination network with two types of isolated, parallel channels, one hydrophilic and the other hydrophobic. PMID- 12722080 TI - The total synthesis of racemic teucvin and 12-epi-teucvin. PMID- 12722081 TI - The dienyl Pauson-Khand reaction. PMID- 12722082 TI - Inverted piano stools: a molecular recognition motif that enforces 1:1 cocrystallization of two diastereomers in the same single crystal. PMID- 12722084 TI - Deprotonation of enoxy radicals: theoretical validation of a 50-year-old mechanistic proposal. PMID- 12722083 TI - 1,4-addition of Grignard reagents to 2,4,6-tri-tert-butyl-1,3,5 triphosphabenzene. PMID- 12722087 TI - Part 1: the history of 19th century neurology and the American Neurological Association. PMID- 12722088 TI - Part 2: history of 20th century neurology: decade by decade. PMID- 12722089 TI - Clinical significance of immunohistochemically detectable epithelial cells in sentinel lymph node and bone marrow in breast cancer. PMID- 12722090 TI - Third look surgery and beyond for appendiceal malignancy with peritoneal dissemination. AB - BACKGROUND AND OBJECTIVES: Second look surgery has been previously studied in patients with recurrent peritoneal dissemination from appendiceal malignancy. However, selection criteria for third, fourth, and subsequent reoperations are not available. METHODS: Five hundred and one patients with epithelial peritoneal surface malignancy of appendiceal origin underwent treatment with cytoreductive surgery and intraperitoneal chemotherapy during an 18-year period. Forty-five of these patients (8.9%) underwent three or more operative interventions. A critical statistical analysis of the impact of selected clinical features on survival was performed from a prospective database. RESULTS: Overall 5-year survival of these 45 patients was 70%. Five- and ten-year survival rates for patients with three interventions were 60 and 48%, for four interventions were 78 and 36%, and for five or more interventions were 100 and 80%. Sites of recurrent disease, histopathologic type of tumor, and lymph node status had no impact on survival. A complete second and third cytoreduction was associated with an improved 5-year survival rate. CONCLUSIONS: Prolonged survival in patients with three or more reoperations was significantly associated with a complete cytoreduction. However, after four or more interventions, the effects of tumor biology may predominate. Although 5-year survival is likely with multiple reoperations, prolonged follow up shows that cancer cure is a rare event. PMID- 12722092 TI - Factors predictive of survival for esophageal carcinoma treated with preoperative radiotherapy with or without chemotherapy followed by surgery. AB - PURPOSE: To evaluate parameters that may influence prognosis in patients treated with preoperative radiotherapy (RT) or chemoradiation. METHODS AND MATERIALS: One hundred seventy-six patients with esophageal carcinoma received preoperative radiotherapy (45 patients) or chemoradiation (131 patients). Forty-three received no surgery (NS), 32 had exploratory surgery (ES), and 101 received definitive surgery (DS). RESULTS: Five-year cause-specific survival and absolute survival rates were overall, 19% and 16%; NS group, 0% and 0%; ES group, 3% and 3%; DS group, 30% and 26%. On univariate analysis, definitive surgery (P < 0.0001), tumor size less than 5 cm (P < 0.0001), and chemotherapy (P = 0.0015) were significant predictors of improved cause-specific survival. Cause-specific survival was 51% for tumors /=6 cm (n = 86) survived. Multivariate analysis of the DS group showed complete or partial pathologic response (P = 0.0001), chemotherapy (P = 0.0026), and overall treatment time less than 3 months (P = 0.0405) significantly predicted improved cause-specific survival. Tumor <5 cm was marginally significant (P = 0.0515). CONCLUSION: Patients who undergo preoperative chemoradiation and definitive surgery have improved survival. PMID- 12722093 TI - Clinical spectrum of primary mediastinal tumors: a comparison of adult and pediatric populations at a single Japanese institution. AB - BACKGROUND AND OBJECTIVES: The purpose of this study was to review our Japanese institutional experience of mediastinal tumors and to compare differences in the clinical spectrum between adults and children. METHODS: We retrospectively reviewed the records of 806 patients (676 adults and 130 children) between 1951 and 2000 at our institution. RESULTS: There were 244 thymomas (36%), 106 germ cell tumors (16%), 95 congenital cysts (14%), 82 lymphomas (12%), 76 neurogenic tumors (11%), and 24 thyroid tumors (4%) in adult patients. There were 60 neurogenic tumors (46%), 24 germ cell tumor (19%), 17 lymphoma (13%), 10 congenital cysts (8%), and 5 thymomas (4%) in the pediatric patients. There was a significant higher incidence of neurogenic tumors and a lower incidence of thymomas and thyroid tumors in children than adults. The most common location was the anterior compartment (68%) in adults and the posterior compartment in children (52%). The prevalence of malignancy in pediatric patients was lower than adults (37% vs. 47%; P < 0.05). Symptoms due to compression or direct invasion into adjacent structures may have caused the higher incidence of dyspnea or respiratory distress in the pediatric group compared to the adults (15.4% vs. 4.0%; P < 0.001). There were fewer incidences of facial edema in children compared to adults (2.6% vs. 10.7%; P < 0.02). Emergent operations were required more often in the pediatric group than the adult group. CONCLUSIONS: We demonstrated definite differences in histologic distribution, location, and symptomatology in mediastinal tumors between adult and pediatric populations, which should be considered in the evaluation and planning of a therapeutic modality for mediastinal tumors. PMID- 12722094 TI - Myocutaneous versus thoraco-abdominal flap cover for soft tissue defects following surgery for locally advanced and recurrent breast cancer. AB - BACKGROUND AND OBJECTIVES: Radical surgical extirpation in advanced breast cancer patients produces large defects that may not be suitable for primary closure. The primary aim in such cases is to achieve an adequate soft tissue cover expeditiously. Various methods, including skin graft, omentum, random flaps, and myocutaneous (MC) flaps, have been tried in the past; however, there is no consensus regarding the method of choice in such patients. METHODS: A retrospective analysis of the breast cancer database of a single surgical unit was performed to find out the incidence of advanced breast cancer patients requiring an additional surgical procedure for soft tissue cover. A comparative analysis of two major groups, "MC flaps" and "thoraco-abdominal (TA) flaps," was performed in relation to operative time, blood loss, morbidity, hospital stay, and final outcome. RESULTS: Forty (12.7%) out of 315 advanced breast cancer patients undergoing surgery required additional surgical procedures for soft tissue cover. MC flaps were used in 18 patients and TA flaps were used in 22 patients. The mean blood loss (192 +/- 77 ml vs. 40 +/- 19 ml), operating time (110 +/- 20 minutes vs. 35 +/- 7 minutes), and hospital stay (10 +/- 4 days vs. 5 +/- 2 days) were significantly less in the TA flap group. CONCLUSIONS: The results of the current study show a significant difference in the blood loss, operating time, and hospital stay between patients requiring TA and MC flap for soft tissue cover following mastectomy. In view of its simplicity and better results, we recommend that the TA flap should be used as a first option flap in advanced breast cancer patients requiring skin cover. PMID- 12722095 TI - Laparoscopic radiofrequency ablation of the liver in proximity to major vasculature: effect of the Pringle maneuver. AB - BACKGROUND AND OBJECTIVES: The Pringle maneuver has been shown to increase ablation size during radiofrequency ablation (RFA). Efficacy of laparoscopic Pringle in proximity to major vasculature has not been well described. Laparoscopic RFA was performed in proximity to major hepatic vessels to examine effects of the Pringle on ablation size and vascular damage. METHODS: Laparoscopic RFA was performed in 10 pigs. Each underwent ablation of a peripheral site, and sites adjacent to the portal and hepatic veins. Ultrasound was used to position the RFA adjacent to vascular structures. US flow characteristics verified occlusion of blood flow. Five pigs underwent laparoscopic RFA with Pringle and five underwent laparoscopic RFA alone. Animals were then sacrificed for gross and microscopic evaluation. RESULTS: Peripheral, hepatic, and portal vein ablations showed no significant differences in volume between non-Pringle and Pringle lesions, though the median ablation volume for the peripheral site in the Pringle group was approximately twice that of the non Pringle group. Pringle group overall median time to target temperature was significantly shorter (P = 0.047). Histologic examination revealed no evidence of endothelial damage or thermal-induced intravascular thrombosis of the hepatic or portal veins. CONCLUSIONS: Laparoscopic RFA with Pringle in proximity to major vascular structures does not significantly increase ablation size, or cause acute vascular damage. Further studies are necessary to determine the utility of the Pringle in proximity to major intrahepatic blood vessels. PMID- 12722097 TI - Radioguided excision of the nonpalpable breast cancer and simultaneous sentinel lymphnode biopsy using a single radiopharmaceutical: an original approach to accurate administration of the blue dye. PMID- 12722096 TI - Budding as a useful prognostic marker in pT3 well- or moderately-differentiated rectal adenocarcinoma. AB - BACKGROUND: Budding along the invasive margin is reported to be associated with greater malignant potential in colorectal adenocarcinoma. We examined the prognostic significance of budding in patients with pT3 rectal carcinoma, particularly in comparison to other routine pathological findings. METHODS: Surgically resected specimens from 83 well- or moderately-differentiated pT3 rectal adenocarcinomas were studied. All resections were curative (R0), and the median postoperative follow-up was 47 months. We examined the presence of budding according to Morodomi's criteria, using hematoxylin-eosin stained sections. RESULTS: Budding was found in 48 patients (57.9%). The incidence of budding was significantly higher in lesions displaying lymphatic invasion and lymph-node metastasis than in those without (P < 0.0001). Both local recurrence and liver metastasis were more frequent in patients with budding-positive lesions (P < 0.002 and 0.02, respectively). Moreover, postoperative survival was significantly decreased in patients with budding-positive lesions than in those with budding negative lesions (cumulative 5-year survival rate were 51.8 and 85.0%, respectively: P < 0.002). Multivariate proportional hazard model revealed that the presence of budding was the only significant co-factor of postoperative survival. CONCLUSIONS: Budding is a pathological marker suggesting high malignant potential and decreased postoperative survival in patients with well- or moderately-differentiated pT3 rectal adenocarcinoma. PMID- 12722098 TI - Siting of drains after surgery. PMID- 12722100 TI - Harvesting a free jejunal graft with a small skin incision. PMID- 12722101 TI - Neuroepithelial cells and associated innervation of the zebrafish gill: a confocal immunofluorescence study. AB - Peripheral chemoreceptors responsive to hypoxia have been well characterized in air-breathing vertebrates, but poorly in water-breathers. The present study examined the distribution of five populations of neuroepithelial cells (NECs), putative O(2) chemoreceptors, and innervation patterns in the zebrafish gill using whole-mounts and confocal immunofluorescence. Nerve bundles and fibers of the gill were labeled with zn-12 (a zebrafish-specific neuronal marker) and SV2 antisera and NECs were characterized by serotonin (5-HT) immunoreactivity (IR), SV2-IR and the purinoceptor P2X(3)-IR. A zn-12-IR nerve bundle extended the length of the gill filament and gave rise to a nerve plexus surrounding the efferent filament artery (eFA) and a rich network of fibers that innervated both serotonergic and nonserotonergic NECs of the filament and lamellar epithelium. Three populations of serotonergic, SV2-IR neurons intrinsic to the gill filaments are described, one of which provided innervation to NECs of the filament epithelium. Degeneration of nerve fibers in gill arches maintained in explant culture for 2 days revealed the extrinsic origin of nerve fibers of the plexus and lamellae and the innervation of filament NECs by both intrinsic and extrinsic fibers. Intrinsic innervation surrounding the eFA survived in explant cultures, suggesting a mechanism of local vascular control within the gill. In addition, NECs survived in explants after degeneration of extrinsic nerve fibers. Thus, NECs of the zebrafish gill are organized in a manner reminiscent of O(2) chemoreceptors of mammalian vertebrates, suggesting a role in respiratory regulation. PMID- 12722103 TI - Differential expression of the Na+-Ca2+ exchanger transcripts and proteins in rat brain regions. AB - In the central nervous system (CNS), the Na(+)-Ca(2+) exchanger plays a fundamental role in controlling the changes in the intracellular concentrations of Na(+) and Ca(2+) ions. These cations are known to regulate neurotransmitter release, cell migration and differentiation, gene expression, and neurodegenerative processes. In the present study, nonradioactive in situ hybridization and light immunohistochemistry were carried out to map the regional and cellular distribution for both transcripts and proteins encoded by the three known Na(+)-Ca(2+) exchanger genes NCX1, NCX2, and NCX3. NCX1 transcripts were particularly expressed in layers III-V of the motor cortex, in the thalamus, in CA3 and the dentate gyrus of the hippocampus, in several hypothalamic nuclei, and in the cerebellum. NCX2 transcripts were strongly expressed in all hippocampal subregions, in the striatum, and in the paraventricular thalamic nucleus. NCX3 mRNAs were mainly detected in the hippocampus, in the thalamus, in the amygdala, and in the cerebellum. Immunohistochemical analysis revealed that NCX1 protein was mainly expressed in the supragranular layers of the cerebral cortex, in the hippocampus, in the hypothalamus, in the substantia nigra and ventral tegmental area, and in the granular layer of the cerebellum. The NCX2 protein was predominantly expressed in the hippocampus, in the striatum, in the thalamus, and in the hypothalamus. The NCX3 protein was particularly found in the CA3 subregion, and in the oriens, radiatum, and lacunoso-moleculare layers of the hippocampus, in the ventral striatum, and in the cerebellar molecular layer. Collectively, these results suggest that the different Na(+)-Ca(2+) exchanger isoforms appear to be selectively expressed in several CNS regions where they might underlie different functional roles. PMID- 12722102 TI - Dlx-1 and Dlx-2 expression in the adult mouse brain: relationship to dopaminergic phenotypic regulation. AB - Expression of the homeodomain-containing transcription factors Dlx-1 and Dlx-2 in the lateral (LGE) and medial (MGE) ganglionic eminences, subpallial embryonic structures, is required for generation of telencephalic interneurons. LGE- and MGE-derived progenitors migrate and populate a number of forebrain structures, including the cortex, hippocampus, and olfactory bulb (OB). Previous reports focusing on embryogenesis of telencephalic neurons in Dlx-1 and Dlx-2 null mice suggested a specific role for these genes in expression of the OB dopamine (DA) phenotype. We have investigated whether these genes also are expressed in adult brain, especially in those pallial derivatives, such as the OB, hippocampus, and possibly cortex, where neurogenesis continues in adults. With a highly sensitive, nonradioactive in situ hybridization technique and both DLX-2 and pan DLX antisera, widespread expression of both genes was found in adult mouse fore- but not mid- or hindbrain. The adult unilateral naris closure paradigm was employed to establish a causative role for Dlx in regulating tyrosine hydroxylase (TH) expression; TH is the first enzyme in DA biosynthesis. TH mRNA, but not Dlx expression, was significantly down-regulated in the OB ipsilateral to closure. These findings suggest that Dlx-1 and -2 do not play a direct role in DA phenotypic differentiation and TH gene regulation in adult OB. The widespread expression of Dlx mRNA and protein in the adult brain suggests that these genes may have additional roles in mature animals. PMID- 12722104 TI - Localization of [3H]nicotine, [3H]cytisine, [3H]epibatidine, and [125I]alpha bungarotoxin binding sites in the brain of Macaca mulatta. AB - We determined the localization of [(3)H]nicotine, [(3)H]cytisine, [(3)H]epibatidine, and [(125)I]alpha-bungarotoxin binding sites in the brain of rhesus monkey by means of receptor autoradiography. The labelings by [(3)H]nicotine, [(3)H]cytisine, and [(3)H]epibatidine were highly concordant, except for epibatidine. Layer IV of some cortical areas, most thalamic nuclei, and presubiculum displayed high levels of labeling for the three ligands. Moderate levels of binding were detected in the subiculum, the septum, and the mesencephalon. Low levels were present in layers I-II and VI of the cortex, the cornu Ammonis, the dentate gyrus, and the amygdala. In addition, the level of epibatidine labeling was very high in the epithalamic nuclei and the interpeduncular nucleus, whereas labeling by nicotine and cytisine was very weak in the same regions. The distribution of [(125)I]alpha-bungarotoxin binding differed from the binding of the three agonists. The labeling was dense in layer I of most cortical areas, dentate gyrus, stratum lacunosum-moleculare of CA1 field, several thalamic nuclei, and medial habenula. A moderate labeling was found in layers V and VI of the prefrontal and frontal cortices, layer IV of primary visual cortex, amygdala, septum, hypothalamus, and some mesencenphalic nuclei. A weak signal was also detected in subiculum, claustrum, stratum oriens, and stratum lucidum of cornu Ammonis and also in some mesencephalic nuclei. The distribution of nicotine, cytisine, and epibatidine bindings corresponds broadly to the patterns observed in rodents, with the marked exception of the epithalamus. However, in monkey, those distributions match the distribution of alpha2 messenger RNA, rather than that of alpha4 transcripts as it exists in rodent brains. The distribution of the binding sites for alpha-bungarotoxin is larger in the brain of rhesus monkeys than in rodent brain, suggesting a more important role of alpha7 receptors in primates. PMID- 12722105 TI - Ectopic corticospinal tract and corticothalamic tract neurons in the cerebral cortex of yotari and reeler mice. AB - Reeler and yotari mice, which are mutant for Reelin or Dab1, respectively, show disorders of cerebral cortical lamination. We injected horseradish peroxidase (HRP) into the upper lumbar enlargement to label corticospinal tract (CST) neurons and wheat germ agglutinin-conjugated HRP (WGA-HRP) into the ventral lateral nucleus of the thalamus to label corticothalamic tract (CTT) neurons in both 19-day-old yotari and reeler mice with the aim of discovering whether or not they show differences in the distribution pattern of layer V or layer VI neurons. Similar injections of tracers were made in normal controls. HRP-labeled CST neurons, which were exclusively distributed in layer V of the normal cortex, were radially scattered in the cortex of both mutants, but those in reeler were more deeply distributed than in yotari. WGA-labeled CTT neurons, which were mainly located in layer VI in the normal cortex, were superficially distributed just beneath the pia mater in both reeler and yotari cortex. The present quantitative study shows that the distribution pattern of layer V neurons, but not layer VI neurons, differs between reeler and yotari mice, suggesting that the Reelin and Dab1 proteins may play different roles in the migration and cell positioning of layer V neurons. PMID- 12722107 TI - Morphology, axonal projection pattern, and responses to optic nerve stimulation of thalamic neurons in the fire-bellied toad Bombina orientalis. AB - Intracellular recording and biocytin labeling were carried out in the fire bellied toad Bombina orientalis to study the morphology and axonal projections of thalamic (TH) neurons and their responses to electrical optic nerve stimulation. Labeled neurons (n = 142) were divided into the following groups: TH1 neurons projecting to the dorsal striatum; TH2 neurons projecting to the amygdala, nucleus accumbens, and septal nuclei; TH3 neurons projecting to the medial or dorsal pallium; TH4 neurons with projections ascending to the dorsal striatum or ventral striatum/amygdala and descending to the optic tectum, tegmentum, and rostral medulla oblongata; TH5 neurons with projections to the tegmentum, rostral medulla oblongata, prectectum, or tectum; and TH6 neurons projecting to the hypothalamus. TH1 neurons are found in the central, TH2 neurons in the anterior and central, TH3 neurons in the anterior dorsal nucleus, and TH4 and TH5 neurons in the posterior dorsal or ventral nucleus. Neurons with descending projections arborize in restricted parts of retinal afferents; neurons with ascending projections do not substantially arborize within retinal afferents. At electrical optic nerve stimulation, neurons in the ventral thalamus respond with excitation at latencies of 10.8 msec; one-third of them follow repetitive stimulation and possibly are monosynaptically driven. Neurons in the dorsal thalamus respond mostly with inhibition at latencies of 42.3 msec and are polysynaptically driven. This corroborates the view that neurons in the dorsal thalamus projecting to the telencephalon receive no substantial direct retinal input and that the thalamopallial pathway of amphibians is not homologous to the mammalian retinogeniculocortical pathway. PMID- 12722106 TI - Synaptic connections of starburst amacrine cells and localization of acetylcholine receptors in primate retinas. AB - Starburst amacrine cells in the macaque retina were studied by electron microscopic immunohistochemistry. We found that these amacrine cells make a type of synapse not described previously; they are presynaptic to axon terminals of bipolar cells. We also confirmed that starburst amacrine cells are presynaptic to ganglion cell dendrites and amacrine cell processes. In order to determine the functions of these synapses, we localized acetylcholine receptors using a monoclonal antibody (mAb210) that recognizes human alpha3- and alpha5-containing nicotinic receptors and also antisera against the five known subtypes of muscarinic receptors. The majority of the mAb210-immunoreactive perikarya were amacrine cells and ganglion cells, but a subpopulation of bipolar cells was also labeled. A subset of bipolar cells and a subset of horizontal cells were labeled with antibodies to M3 muscarinic receptors. A subset of amacrine cells, including those that contain cholecystokinin, were labeled with antibodies to M2 receptors. Taken together, these results suggest that acetylcholine can modulate the activity of retinal ganglion cells by multiple pathways. PMID- 12722108 TI - Distribution of CNTF receptor alpha protein in the central nervous system of the chick embryo. AB - Ciliary neurotrophic factor (CNTF) promotes the survival and differentiation of various neuronal and glial cell populations in the nervous system of vertebrates. In mammals, the ligand-binding alpha-subunit of the CNTF receptor (CNTFRalpha) is expressed in a variety of neuronal populations, including all CNTF-responsive cells. Previous studies suggested that functional differences in the CNTF/CNTF receptor system between chicks and mammals exist. The purpose of the present study was to examine the temporal and spatial expression pattern of the chick CNTFRalpha protein during CNS development. Receptor expression was detectable by immunoblotting in all CNS areas tested but showed area-specific developmental regulation. Interestingly, two variants of CNTFRalpha, 69 and 65 kD, were identified by immunoblotting with a shift from the higher to the lower molecular mass species occurring during development. Immunoreactivity for CNTFRalpha protein was preferentially observed in neuropil and white matter structures of the developing CNS while neuronal somata generally appeared unlabeled. For example, expression was observed in the olfactory system, in the telencephalon, in parts of the somatosensory system, in components of the tectofugal pathway, in the cerebellum, and in auditory brainstem nuclei. Fiber tracts that exhibit CNTFRalpha immunoreactivity were the lateral forebrain bundle, occipitomesencephalic tract, quintofrontal tract, and vestibular nerve. Our study identifies potential new targets of a chick CNTF-related molecule and reveals significant regional differences of CNTFRalpha protein expression between chick and mammals. These results suggest that the CNTF receptor performs distinct developmental functions in different animals. PMID- 12722109 TI - Determinants of the exclusion zone in dopaminergic amacrine cell mosaics. AB - A fundamental organizing feature of the retina is the presence of regularly spaced distributions of neurons, yet we have little knowledge of how this patterning emerges during development. Among these retinal mosaics, the spatial organization of the dopaminergic amacrine cells is unique: using nearest-neighbor and Vornoi domain analysis, we found that the dopaminergic amacrine cells were neither randomly distributed, nor did they achieve the regularity documented for other retinal cell types. Autocorrelation analysis revealed the presence of an exclusion zone surrounding individual dopaminergic amacrine cells and modeling studies confirmed this organization, as the mosaic could be simulated by a minimal distance spacing rule defined by a broad set of parameters. Experimental studies determined the relative contributions of tangential dispersion, fate determination, and cell death in the establishment of this exclusion zone. Clonal boundary analysis and simulations of proximity-driven movement discount tangential dispersion, while data from bcl-2 overexpressing mice rule out feedback-inhibitory fate-deterministic accounts. Cell death, by contrast, appears to eliminate dopaminergic amacrine cells that are within close proximity, thereby establishing the exclusion zone surrounding individual cells and in turn creating their mosaic regularity. PMID- 12722110 TI - Fourier transform IR spectroscopy study for new insights into molecular properties and activation mechanisms of visual pigment rhodopsin. AB - Fourier transform IR (FTIR) spectroscopy has been successfully applied in recent years to examine the functional and structural properties of the membrane protein rhodopsin, a prototype G protein coupled receptor. Unlike UV-visible spectroscopy, FTIR spectroscopy is structurally sensitive. It may give us both global information about the conformation of the protein and very detailed information about the retinal chromophore and all other functional groups, even when these are not directly related to the chromophore. Furthermore, it can be successfully applied to the photointermediates of rhodopsin, including the active receptor species, metarhodopsin II, and its decay products, which is not expected presently or even in the near future from crystallographic approaches. In this review we show how FTIR spectroscopy has significantly contributed to the understanding of very different aspects of rhodopsin, comprising both structural properties and the mechanisms leading to receptor activation and deactivation. PMID- 12722112 TI - Hydration of hyaluronan polysaccharide observed by IR spectrometry. III. Structure and mechanism of hydration. AB - The results of the analysis of hydration spectra of Na(+) hyaluronan (HA) performed in a companion study are translated in terms of chemical mechanisms. We find that dried HA is characterized by chains having ordered parts of at least 6 disaccharide repeat units that extend over 60 A. The order is mainly due to C3O3H...O5 and C4O4H...O5 hydrogen bonds that hinder rotations around beta(1-4) and beta(1-3) glycoside bonds. Along one chain there are two-thirds of the N-H amide groups and carboxyl groups that are directly hydrogen bonded, with no water intermediate, to form N-H...(-)O-C=O hydrogen bonds, which are collateral to C3O3H...O5 hydrogen bonds. The existence of these N-H...(-)O-C=O bonds is somewhat in opposition to literature descriptions. In this dry state a "water wire" of 4-5 H(2)O molecules, which are anchored on C=O carboxyl groups and hydrating the Na(+) CO(-) ionic group, establishes hydrogen bonds on other hydrophilic groups of the same chain or other chains and remains embedded in HA, even at 104 degrees C. Hydration occurs at low hygrometry around the remaining one-third of the N-H...(-)O-C=O pairs that are not hydrogen bonded. Each of these N-H and (-)O-C=O groups is hydrated by a nanodroplet of some 25 H(2)O molecules that finds other sites for binding and hydrates 2 disaccharide repeat units. At higher hygrometry bigger nanodroplets hydrate all hydrophilic sites. PMID- 12722111 TI - Hydration of hyaluronan polysaccharide observed by IR spectrometry. II. Definition and quantitative analysis of elementary hydration spectra and water uptake. AB - We recorded a series of spectra of sodium hyaluronan (HA) films that were in equilibrium with their surrounding humid atmosphere. The hygrometry of this atmosphere extended from 0 to 0.97% relative humidity. We performed a quantitative analysis of the corresponding series of hydration spectra that are the difference spectra of the film at a defined hygrometry minus the spectrum of the dried film (hygrometry = 0). The principle of this analysis is to use this series of hydration spectra to define a limited number (four) of "elementary hydration spectra" over which we can decompose all hydration spectra with good accuracy. This decomposition, combined with the measurements of the numbers of H(2)O molecules at the origin in these elementary hydration spectra of the three characteristic vibrational bands of H(2)O, allowed us to calculate the hydration number under different relative humidity conditions. This number compares well with that determined by thermogravimetry. Furthermore, the decomposition defines for each hygrometry value which chemical mechanisms represented by elementary hydration spectra are active. This analysis is pursued by determining for the elementary hydration spectra the number of hydrogen bonds established by each of the four alcohol groups found in each disaccharide repeat unit before performing the same analysis for amide and carboxylate groups. These results are later utilized to discuss the structure of HA at various stages of hydration. PMID- 12722113 TI - Interaction of antitumoral 9-aminoacridine drug with DNA and dextran sulfate studied by fluorescence and surface-enhanced Raman spectroscopy. AB - Fluorescence spectroscopy and surface-enhanced Raman spectroscopy are applied to study the interaction of the drug 9-aminoacridine (9AA) with DNA and dextran sulfate. The effect of the electrostatic interaction between the positively charged 9AA and negatively charged groups in relation to the excimer or exciplex emission is investigated. The exciplex emission of 9AA is connected to the intercalation of this drug between nucleic base residues. The importance of negative groups in this interaction is evaluated by using dextran and dextran sulfate as model polymers. The existence of negative charges seems to induce an increase of the drug concentration in the vicinity of the polymers. The role of electrostatic attraction in the 9AA dimerization is confirmed by the excimer emission of 9AA in the presence of dextran sulfate. In the case of DNA, the phosphate groups may induce the drug approach to the DNA chain, but the exciplex fluorescence emission could be due to a charge transfer between the drug and adenine-rich sequences of DNA. PMID- 12722114 TI - FT-Raman spectroscopy as diagnostic tool of Congo red binding to amyloids. AB - Chorion is the major component of silkmoth eggshell. More than 95% of its dry mass consists of the A and B families of low molecular weight structural proteins, which have remarkable mechanical and chemical properties protecting the oocyte and developing embryo from environmental hazards. We present data from FT Raman spectroscopy of silkmoth chorion and amyloid-like fibrils formed from peptide analogues of chorion proteins, both unstained and stained by Congo red. The results show that FT-Raman spectroscopy is not a straightforward diagnostic tool for the specific interactions of Congo red with amyloids: a dilute aqueous solution of the Congo red dye at pH 5.5 and a thin solid film of the dye cast from this solution exhibit the same "diagnostic" Raman shifts relative to the neat Congo red dry powder as do amyloid fibrils formed from peptide analogues of chorion proteins stained by Congo red. An important consequence of this finding is that these shifts of the Raman active modes of Congo red are probably due to the formation of supramolecular dye aggregates in the presence of water. Therefore, this is not an appropriate diagnostic test for Congo red binding to amyloids. PMID- 12722115 TI - IR spectra of cytochrome c denatured with deuterated guanidine hydrochloride show increase in beta sheet. AB - Attenuated total reflectance Fourier transform IR (ATR-FTIR) spectra are obtained for horse heart ferricytochrome c in solutions of 0-7M guanidine hydrochloride and deuterated guanidine hydrochloride. Substitutions of deuterium for hydrogen in both the denaturant and protein provide resolvable amide I spectra over a wide range of denaturant concentrations. Deuteration enhances the ability to measure the true protein IR spectrum in the amide I region in which the secondary structure can be deduced, because spectra in D(2)O are less prone to spectral distortion upon background denaturant subtraction than spectra in H(2)O. Other investigators studying equilibrium unfolded cytochrome c were limited to guanidine concentrations below 3.0M because of detector saturation. Detector saturation is avoided with the use of ATR-FTIR spectroscopy, allowing one to obtain protein spectra at high denaturant concentrations. Second derivative spectra of samples show reductions in alpha helix and increases in beta sheet at high denaturant concentrations, contrary to expectations of finding primarily a random coil secondary structure. Using this new technique, the protein was estimated to consist of 51% beta sheet and only 15% random coil in the presence of 6.6M deuterated guanidine hydrochloride. PMID- 12722116 TI - Mollaret's meningitis: cytopathologic analysis of fourteen cases. AB - Mollaret's meningitis (MM) is a rare disease of benign nature characterized by recurrent episodes of aseptic meningitis. Cerebrospinal fluid (CSF) examination remains the sole diagnostic modality. Eighteen CSF samples from 14 patients were studied along with the clinical data. Specimens were prepared by cytocentrifugation and Millipore filtration and were stained with Diff-Quik and Papanicolaou stains. Eight patients were men and six were women, with an age range of 17-74 yr (mean age 37 yr). Most common clinical presentation was recurrent episodes of headaches and photophobia followed by a sustained mild fever lasting 5-7 days. The CSF showed markedly increased cellularity with pleocytosis. The differential count showed predominant monocytosis ranging from 84% to 100% (mean 96). In our series, two patients had herpes simplex virus type 2 (HSV-2) DNA detected by polymerase chain reaction (PCR) in the CSF. The monocytes were seen predominantly singly, but three cases showed a strong tendency to aggregate in small groups. Phenotypically, these cells had bean shaped bilobed nuclei as well as multiple deep nuclear clefts depicting the so called "footprint" appearance. In four cases, multiple blunt-tipped cytoplasmic pseudopods were noted. Degenerated monocytes with the appearance of the so-called "ghost cells" were noted in one-half of the cases. Background cells were mostly small mature lymphocytes; however, one-half of cases showed a significant amount of plasma cells and/or polymorphonuclear leukocytes (PMNs). Lysed blood with hemosiderin-laden macrophages and numerous leptomeningeal cells were seen in two cases. CSF examination of MM presents a spectrum of cytomorphologic features. When interpreted in light of the appropriate clinical setting. the latter, although nonspecific, provides an accurate diagnosis. The differential diagnosis includes various degenerative, inflammatory/infectious, and lymphoproliferative disorders of the central nervous system. PMID- 12722117 TI - Fine-needle aspiration of synovial sarcoma: criteria for diagnosis: retrospective reexamination of 37 cases, including ancillary diagnostics. A Scandinavian Sarcoma Group study. AB - The cytologic criteria of synovial sarcoma in fine-needle aspirates were defined by a retrospective examination of 37 primary tumors. Irrespective of subtype, a typical pattern at low power was found, provided the yield was rich. The typical pattern was a mixture of dispersed cells with the presence of striped nuclei and cell-tight tumor tissue fragments with irregular borders. Often a branching network of vessels was present in the fragments, imitating a true vascular tumor. Except in poorly differentiated synovial sarcomas, the tumor cells were, small to medium in size, with rounded, ovoid, or fusiform bland nuclei with inconspicuous nucleoli. In the biphasic variant, small glandular- or acinar-like structures were present, although not in all cases. In the poorly differentiated type, however, the cellular pleomorphism was marked with the presence of cells with irregular nuclei and rhabdomyoblast-like cells, corresponding to the pleomorphic variant. The Ewing's sarcoma-like and the atypical spindle cell variants of poorly differentiated synovial sarcoma were not diagnosed in the material. An unequivocal diagnosis of sarcoma is possible when the yield is rich. However, ancillary diagnostics are necessary for a correct diagnosis, to avoid important pitfalls, such as other sarcomas with bland tumor cells and vessel-rich tumor fragments, in particular, solitary fibrous tumor and true hemangiopericytoma. Electron microscopic and/or molecular genetic analyses were better diagnostic adjuncts than immunocytochemistry. PMID- 12722118 TI - Cytology of the central zone of the prostate. AB - The prostate has three anatomical regions: the peripheral, transition, and central zones (CZ). The CZ has distinct histological features, but its cytological morphology has not been described. This study was done on surgical specimens to ensure that samples were representative of the CZ, and that no prostatic intraepithelial neoplasia (PIN) or cancer contaminated the smears. An incision was made in the CZ of 51 prostatectomy specimens, and cells were scraped from cut surfaces. After exclusion of samples contaminated by PIN or cancer or with poor cell yield, 39 Giemsa-stained smears remained for analysis. Large branching epithelial sheets with geographic architecture and crowded nuclei were seen in 97% of smears. Epithelial clusters with elongated palisaded nuclei were identified in 80% of cases, but were always a minor component. Visible nucleoli (97%), cytoplasmic vacuoles (97%), and smooth muscle cells in the background (95%) were common. Blue-green cytoplasmic granules resembling seminal vesicle pigment were seen in 97%. Magenta-colored cytoplasmic pigment, similar to granules seen in other regions of the prostate, was found in 74%. Recognition of CZ epithelium as a benign constituent of prostate cytology is important because elongated cells, crowded nuclei, and visible nucleoli may otherwise be misinterpreted as PIN or cancer. PMID- 12722119 TI - Cervicovaginal (Papanicolaou) smear findings in patients with malignant mixed Mullerian tumors. AB - Malignant mixed Mullerian tumor is a rare neoplasm that occurs most frequently in elderly patients. It is characterized by a mixture of malignant epithelial and sarcomatous components. Little has been published about Papanicolaou smear findings pertaining to malignant mixed Mullerian tumors. We present our experience, with an emphasis on cytologic detail. Nine patients (median age, 65 yr) met our study criteria. All available smears and surgical specimens were reviewed. Four smears were positive for malignancy, with a sensitivity of 44% (3 adenocarcinoma, and 1 squamous-cell carcinoma, small-cell type). The results of our study showed that Papanicolaou smear findings pertaining to malignant mixed Mullerian tumors are seen in patients with advanced-stage disease with involvement of the lower uterine segment or cervix. The usual finding is large numbers of high-grade epithelial malignant cells in a necrotic background. The mesenchymal component rarely sheds cells visible on Papanicolaou smear. PMID- 12722120 TI - Scrape cytology of the ovaries: potential role in intraoperative consultation of ovarian lesions. AB - Intraoperative diagnosis of ovarian lesions can be achieved by gross examination with the help of frozen sections and/or cytologic examination. Various cytologic techniques, including imprint, fine-needle aspiration, and scrape, may be used. In this study, we evaluated the application of scrape cytology in the diagnosis of ovarian lesions occurring during a 16-mo period at one institution. The cytologic diagnosis was primarily based on findings in air-dried, Diff-Quik stained smears in correlation with clinical and intraoperative findings. In total, 131 histologically proven ovarian lesions, including 13 nonneoplastic lesions, 47 benign tumors, 17 epithelial tumors of low malignant potential (LMP), and 54 malignant tumors (35 primary, 1 leukemia, and 18 metastases), were cytologically examined. The accuracy of scrape cytology was 95% in the benign, 47% in the LMP, and 98% in the malignant group. In the LMP group, the false negative rate was 12%, while the deferred rate and false-positive rate were 24% and 18%, respectively. There was no misdiagnosis between the benign and malignant categories. The histologic subtypes were correctly predicted in 78% of cases. There were limitations of scrape cytology in the diagnosis of LMP and mucinous tumors, which require histologic architectural evaluation and adequate histologic sampling. Scrape cytology is a simple, rapid, and inexpensive adjunctive technique that should be used in intraoperative consultation for ovarian lesions. PMID- 12722121 TI - Fine-needle aspiration cytology in lesions related to ornamental body procedures (skin tattooing, intraoral piercing) and recreational use of drugs (intranasal route). AB - Body adornment through tattooing and body piercing and the use of recreational drugs are on the increase, producing a variety of secondary lesions, the etiology of which often remains undetected, as the medical community is not yet aware of the extent of the morbidity of such procedures and practices. Three cases are presented, which underscore the problem and also the role that fine-needle aspiration (FNA) can play in clarifying the etiology of such lesions. Two of these cases were lymphadenopathies, one secondary to tattooing and the other to tongue piercing, while the third was a deep intranasal lesion, which in all probability had resulted from intranasal use of recreational drugs. Although the clinical diagnosis of these lesions was problematic, the FNA performed by a pathologist, by associating the cytologic findings with the corresponding clinical setting, was quite indicative of their relation to the aforementioned procedures or practices. PMID- 12722122 TI - Cytologic differential diagnosis of myxoid and mucinous neoplasms of the sacrum and parasacral soft tissues. AB - A diagnostically important group of lesions involving the sacrum, spinal canal, and parasacral soft tissues is characterized by a myxoid or mucinous background in cytologic smears. This group of myxoid/mucoid neoplasms includes chordoma, myxopapillary ependymoma, metastatic mucinous adenocarcinoma, and extraskeletal myxoid chondrosarcoma. Despite the similarity of the background substance, each neoplasm within this differential diagnosis has a characteristic composite set of morphologic and immunophenotypic features. Because many of these masses are not easily surgically biopsied, fine-needle aspiration (FNA) is often used for their diagnosis. The private consultation records of the author and the cytology files of the University of Utah Department of Pathology were searched for all lesions arising in or around the sacrum. These cases were reviewed to determine which had a myxoid/mucinous background. Fourteen neoplasms were found and comprise the study set. Four of these cases had cell block material in addition to Diff-Quik stained smears; a panel of antibodies, including cytokeratin, glial fibrillary acid protein (GFAP), S-100 protein, and carcinoembryonic antigen (CEA), was performed on the cell block material. The smears were evaluated for cytologic features, including the presence of rosette-like structures, physaliphorous cells, gland-like structures, chondroid fragments, "signet ring" and "goblet" cells, as well as the character of the myxoid/mucinous background substance. The cases included one myxopapillary ependymoma, 10 chordomas, two mucinous adenocarcinomas of colonic or gastric origin, and one myxoid chondrosarcoma. Physaliphorous cells were found to be highly specific for chordoma, while a fibrillary myxoid stroma containing cells with elongated cytoplasmic processes and cells lying in a rosette-like pattern around central cores of myxoid to fibrillary stroma were highly characteristic of myxopapillary ependymoma. Fragments of a myxoid/chondroid matrix with lacunar-like spaces strongly supported the diagnosis of myxoid chondrosarcoma. "goblet" or "signet ring" cells with a single distinct vacuole favored mucinous adenocarcinoma. There appear to be sufficient cytomorphologic features present within the FNA smears and cell block material to allow cytologic separation of the more common myxoid and mucinous neoplasms involving the sacral and parasacral tissues. PMID- 12722123 TI - Cytologic diagnosis of fibrous hamartoma of infancy: a case report. AB - Fibrous hamartoma of infancy (FHI) is a rare benign lesion in the first 2 years of life with a distinctive histomorphologic appearance. There were only two descriptions of this entity in the cytology literature. In this article, the detailed clinical profile and cytologic features of a case of FHI is described in a 6-month-old male infant who presented with a nodular mass in the anterior axillary fold. Fine-needle aspiration (FNA) cytology smears showed an admixture of adipose tissue fragments, discretely lying, and clusters of fibroblastic cells in association with myxoid and collagenous matrix. FNA cytology features along with proper clinical history, radiological findings, family history, and location of the lesion may be helpful to suggest the probable preoperative diagnosis of this lesion. PMID- 12722124 TI - Meningioma with extensive noncalcifying collagenous whorls and glial fibrillary acidic protein expression: new variant of meningioma diagnosed by smear preparation. AB - The purpose of this report is to describe the unique cytological findings of a new recently characterized type of meningioma that has extensive noncalcifying collagenous whorls and glial fibrillary acid protein (GFAP) expression. This new entity, described by Haberler and colleagues, was named whorling sclerosing variant of meningioma. The patient was a 34-yr-old white man with a large tumor in the brainstem. Intraoperative smear preparations showed a tumor with a large number of solid hyaline masses in a loose background and in focal areas tumor cells formed cohesive nests with a somewhat whorling appearance. The histological sections showed a neoplasia composed of innumerable eosinophilic, collagenous, noncalcified round deposits, cuffed by scattered meningothelial tumor cells. The neoplastic cells showed diffuse cytoplasmic reactivity for EMA and vimentin, as well as positivity to GFAP. This is the first cytological description of this new entity in the literature. PMID- 12722125 TI - Interdigitating dendritic reticulum cell tumor of lymph nodes: case report with differential diagnostic considerations. AB - A 66-yr-old man with a history of interdigitating reticulum cell tumor (IDRCT) presented with an enlarging right cervical lymph node suspicious of recurrent disease. He reported no fevers or night sweats and was otherwise asymptomatic at the time of the fine-needle aspiration (FNA). FNA of the lymph node showed a spindle cell lesion with plump, at times deeply convoluted, large nuclei, coarsely granular chromatin, with one or two distinct nucleoli, intimately associated with mature-appearing lymphocytes. A cell block was made and immunohistochemistry was carried out. Immunopositivity for vimentin, S-100, and CD68 and immunonegativity for CD21 and AE1/AE3 were noted. This case report is the first to illustrate the FNA cytomorphology of IDRCT and to discuss the cytologic differential diagnosis of this rare tumor type in lymph nodes. PMID- 12722126 TI - Papanicolaou Society of Cytopathology guidelines for educational notes, disclaimers, and similar comments on reports of cervical cytology specimens. AB - Controversy exists regarding the need for and appropriate form of notes addended to cervical cytology reports. The Guidelines Task Force of the Papanicolaou Society of Cytopathology reviewed the pertinent literature and formulated a list of suggested guidelines to aid practicing cytopathologists in conforming to the National Cancer Institute Bethesda Workshop's consensus recommendations and to fill the requirements of the College of American Pathologists inspections. PMID- 12722127 TI - Maximal efficiency of PAPNET in the diagnosis of infections in cervicovaginal smears. PMID- 12722128 TI - Pharmacological and toxicological properties of Nigella sativa. AB - The seeds of Nigella sativa Linn. (Ranunculaceae), commonly known as black seed or black cumin, are used in folk (herbal) medicine all over the world for the treatment and prevention of a number of diseases and conditions that include asthma, diarrhoea and dyslipidaemia. This article reviews the main reports of the pharmacological and toxicological properties of N. sativa and its constituents. The seeds contain both fixed and essential oils, proteins, alkaloids and saponin. Much of the biological activity of the seeds has been shown to be due to thymoquinone, the major component of the essential oil, but which is also present in the fi ed oil. The pharmacological actions of the crude extracts of the seeds (and some of its active constituents, e.g. volatile oil and thymoquinone) that have been reported include protection against nephrotoxicity and hepatotoxicity induced by either disease or chemicals. The seeds/oil have antiinflammatory, analgesic, antipyretic, antimicrobial and antineoplastic activity. The oil decreases blood pressure and increases respiration. Treatment of rats with the seed extract for up to 12 weeks has been reported to induce changes in the haemogram that include an increase in both the packed cell volume (PCV) and haemoglobin (Hb), and a decrease in plasma concentrations of cholesterol, triglycerides and glucose. The seeds are characterized by a very low degree of toxicity. Two cases of contact dermatitis in two individuals have been reported following topical use. Administration of either the seed extract or its oil has been shown not to induce significant adverse effects on liver or kidney functions. It would appear that the beneficial effects of the use of the seeds and thymoquinone might be related to their cytoprotective and antioxidant actions, and to their effect on some mediators of inflammation. PMID- 12722129 TI - Protective effect of a polyherbal formulation (Immu-21) against cyclophosphamide induced mutagenicity in mice. AB - The object was to evaluate the effects of a polyherbal formulation, Immu-21, against cyclophosphamide (CP)-induced chromosomal aberrations (CA) and micronuclei (MN) in mice. CP alone (40 mg/kg, i.p.) produced classical as well as non-classical chromosomal aberrations in mice, and the incidence of CA was significantly more in the CP treated group when compared with that of the control group. Immu-21, which contains extracts of Ocimum sanctum, Withania somnifera, Emblica officinalis and Tinospora cordifolia, was given at 100 mg/kg, daily, over 7 days, and 30 mg/kg daily over 14 days and inhibited both CP-induced classical and non-classical chromosomal aberrations ( approximately 40%-60% of control). A significant increase in MN was also observed in bone marrow erythrocytes of mice treated with CP, and pretreatment with Immu-21 also significantly reduced these. Cytotoxicity was evaluated by estimating the ratio of polychromatic erythrocytes (PCEs) to normochromatic erythrocytes (NCEs). The present results indicate that chronic treatment with Immu-21 prevented CP-induced genotoxicity in mice. PMID- 12722130 TI - Glutathione S-transferases and malondialdehyde in the liver of NOD mice on short term treatment with plant mixture extract P-9801091. AB - Changes in the concentration of glutathione S-transferases (GSTs) and malondialdehyde (MDA) were assessed in the liver of normal and diabetic NOD mice with and without treatment with the plant extract P-9801091. The plant extract P 9801091 is an antihyperglycaemic preparation containing Myrtilli folium (Vaccinium myrtillus L.), Taraxaci radix (Taraxacum of fi cinale Web.), Cichorii radix (Cichorium intybus L.), Juniperi fructus (Juniperus communis L.), Centaurii herba (Centaurium umbellatum Gilib.), Phaseoli pericarpium (Phaseolus vulgaris L.), Millefoliiherba (Achillea millefolium L.), Mori folium (Morus nigra L.), Valerianae radix (Valeriana of ficinalis L.) and Urticae herba et radix (Urtica dioica L). Hyperglycaemia in diabetes mellitus is responsible for the development of oxidative stress (via glucose auto-oxidation and protein glycation), which is characterized by increased lipid peroxide production (MDA is a lipid peroxidation end product) and/or decreased antioxidative defence (GST in the liver is predominantly an alpha enzyme, which has antioxidative activity). The catalytic concentration of GSTs in the liver was significantly reduced in diabetic NOD mice compared with normal NOD mice (p < 0.01), while the concentration of MDA showed a rising tendency (not significant). The results showed that statistically significant changes in antioxidative defence occurred in the experimental model of short-term diabetes mellitus. A 7-day treatment with P-9801091 plant extract at a dose of 20 mg/kg body mass led to a significant increase in the catalytic concentration of GSTs in the liver of diabetic NOD mice (p < 0.01) and a decrease in MDA concentration (not significant), which could be explained by its antihyperglycaemic effect. PMID- 12722131 TI - Male rat infertility induction/spermatozoa and epididymal plasma abnormalities after oral administration of Kalanchoe gastonis bonnieri natural juice. AB - Natural aqueous crude extracts (NACE) of several Crassulaceae family plants have been applied as a vaginal contraceptive by the populace. The aim of this work was to evaluate the inhibition of fertility in male Wistar rats and some physiological and biochemical changes in spermatozoa and epididymal plasma induced by NACE from Kalanchoe gastonis bonnieri (K. g. b.) (Crassulaceae). The NACE was obtained by mechanic pressure on grinding fresh plant leaves. Sublethal doses (150-300 mg/kg body weight) of NACE were orally administered to adult and fertile male rats daily for 30 days in a search for a contraceptive effect, and physiological and biochemical modifications on sperm cells and cauda epididymal plasma. The toxicity studies revealed that the lethal dose (LD(50)) calculated was 11 g/kg body weight. Sublethal doses induced 50%-100% fertility inhibition, with 100% recovery of fertility 30 days after stopping the treatment. The sperm motility, viability and spermatic density were also significantly decreased (p < 0.001). The outstanding biochemical change observed in the cauda epididymal plasma was a decrease of carnitine concentration. The NACE of K. gastonis contains one substance active on fertility by affecting spermatozoa motility, viability and sperm density with a significantly decreased carnitine and sialic acid (p < 0.001) in the caudal epididymal plasma. PMID- 12722132 TI - Smooth muscle relaxant effect of kaurenoic acid, a diterpene from Copaifera langsdorffii on rat uterus in vitro. AB - The effect of kaurenoic acid, a diterpene isolated from the oleo-resin of the popular medicinal plant Copaifera langsdorffii (Leguminaceae), was analysed on rat uterine muscle responsiveness to various drugs in vitro. Cumulative concentration-response curves to acetylcholine and oxytocin were obtained before and after incubation of uterine segments with up to 160 microm of kaurenoic acid. The maximal contractile response (E(max)) evoked by these agonists was inhibited by kaurenoic acid in a concentration-related manner; at 160 microm, kaurenoic acid depressed the E(max) of oxytocin and acetylcholine by 83% and 91%, respectively. The relaxation caused by kaurenoic acid on oxytocin-induced contraction was unaffected in the presence of tetraethyl ammonium, a compound that blocks the calcium activated potassium channels. It was partially reversed by glibenclamide (10(-5) m), an ATP-sensitive potassium channel blocker. Also, kaurenoic acid at 160 microm concentration was found to inhibit significantly the CaCl(2)-evoked contractile responses in a medium of high potassium and zero calcium. Furthermore, kaurenoic acid was found to relax the sustained tonic contraction induced by acetylcholine, oxytocin, BaCl(2) and KCl in a concentration-dependent way. However, KCl-induced tonic contraction was only weakly inhibited by kaurenoic acid. These data indicate that the diterpene, kaurenoic acid, exerts a uterine relaxant effect acting principally through calcium blockade and in part, by the opening of ATP-sensitive potassium channels. PMID- 12722134 TI - Protective effect of Spirulina on lead induced deleterious changes in the lipid peroxidation and endogenous antioxidants in rats. AB - The present study aims to investigate the protective effect of Spirulina on lead induced changes in the levels of lipid peroxidation and endogenous antioxidants in liver, lung, heart, kidney and brain of rats. Levels of elemental lead were also measured in the organs of rats in all experimental groups. In the liver, lung, heart and kidney of lead-exposed animals, there was a significant (p < 0.001) increase in the lipid peroxidation and a decrease in the levels of endogenous antioxidants. Although, Spirulina did not affect the deposition of lead in organs apart from the brain, simultaneous administration of Spirulina to lead exposed animals significantly (p < 0.001) inhibited lipid peroxidation and restored the levels of endogenous antioxidants to normal. To conclude, Spirulina had a significant effect on scavenging free radicals, thereby protecting the organs from damage caused by the exposure to lead. Further more, Spirulina showed a significant (p < 0.05) decrease in the deposition of lead in the brain. PMID- 12722133 TI - Antibacterial, antiprotozoal and antioxidant activity of five plants used in Izabal for infectious diseases. AB - Methanol and aqueous extracts from fi ve plant species, used in traditional medicine in Guatemala for the treatment of microbial infections, were tested in vitro for their ability to scavenge DPPH, OH(.) and O(2) (-) radicals and to inhibit lipoperoxidation (LPO) in order to establish a relationship between their antioxidant activities and their effects against infectious agents. Acalypha guatemalensis, Ocimum micranthum and Smilax spinosa possessed a significant activity against both the three free radicals assayed and LPO; Guazuma ulmifolia showed effects against DPPH and OH(.). Piper auritum showed no activity. These extracts were also evaluated for antibacterial and antiprotozoal activities. A. guatemalensis showed activity against Pseudomonas aeruginosa; S. spinosa was active against Salmonella typhi, and A. guatemalensis, and S. spinosa against Trypanosoma cruzi or Leishmania spp. PMID- 12722136 TI - Antiangiogenic activity of lupeol from Bombax ceiba. AB - In the search for antiangiogenic agents from medicinal plants used in Vietnam, a methanol extract of the stem barks of Bombax ceiba was found to exhibit a significant antiangiogenic activity on in vitro tube formation of human umbilical venous endothelial cells (HUVEC). Bioactivity-guided fractionation and isolation carried out on this extract afforded lupeol as an active principle. At 50 and 30 microg/mL lupeol showed a marked inhibitory activity on HUVEC tube formation while it did not affect the growth of tumor cell lines such as SK-MEL-2, A549, and B16-F10 melanoma. PMID- 12722135 TI - Anti-inflammatory and smooth muscle relaxant activities of the hydroalcoholic extract and chemical constituents from Amburana cearensis A C Smith. AB - Amburana cearensis A. C. Smith, Fagaceae, is a medicinal plant commonly known as 'cumaru' and used in Northeast Brazil for the treatment of respiratory tract diseases. In the present work, we investigated the anti-inflammatory and smooth muscle relaxant activities of the hydroalcoholic extract (HAE), coumarin (Coum) and fl avonoid fraction (FF) isolated from the trunk barks of Amburana cearensis A. C. Smith. It was shown that HAE (200 and 400 mg/kg), Coum (20 and 40 mg/kg) and FF (40 mg/kg), administered orally, significantly inhibited both leukocyte and neutrophil migrations, in the carrageenan or N-formyl-methyl-leucyl phenylalanine (fMLP)-induced migration in rat peritoneal cavity. The increase in cutaneous vascular permeability induced by serotonin in rats was significantly blocked by HAE (150 mg/kg, i.p.), Coum (5 mg/kg, i.p.) and FF (20 mg/kg, i.p.). However, only HAE blocked the histamine effect on Evans blue extravasation. In the guinea-pig trachea precontracted with carbachol (0.3 microM), histamine (0.1 microM) or KCl (0.1 M), the HAE, Coum and FF evoked a concentration-dependent relaxation in the presence of the three agonists. HAE (100-800 microg/ml) and Coum (4-32 microg/ml) also caused significant relaxation of the rat vas deferens previously contracted with adrenaline, acetylcholine or barium chloride. In addition, HAE, Coum and FF inhibited the histamine and serotonin-induced increase of cutaneous vascular permeability in rats. PMID- 12722137 TI - Antioxidant flavonoids and phenolic acids from leaves of Leea guineense G Don (Leeaceae). AB - From leaves of Leea guineense (Leeaceae) three hydrophilic flavonoids were isolated and identified as quercetin-3'-sulphate-3-O-alpha-L-rhamnopyranoside, quercetin-3,3'-disulphate and a new flavonoid sulphate, quercetin-3,3',4' trisulphate, together with kaempferol, quercetin, quercitrin, mearnsitrin, gallic acid and ethyl gallate. The structures were established by spectroscopic analysis (UV, MS, (1)H-NMR, (13)C-NMR and 2D-NMR). Their antioxidant effect on free radical scavenging was evaluated in the DPPH assay. PMID- 12722138 TI - Cytotoxic and multidrug resistance reversal activity of a vegetable, 'Anastasia Red', a variety of sweet pepper. AB - The vegetable, Anastasia Red, Capsicum annuum L. var. angulosum Mill. (Solanaceae) was successively extracted with hexane, acetone, methanol and 70% methanol, and the extracts were further separated into a total of 21 fractions by silica gel or octadecylsilane (ODS) column chromatography. The biological activities of extracts and fractions were determined. These extracts showed relatively higher cytotoxic activity against two human oral tumor cell lines (HSC 2, HSG) than against normal human gingival fibroblasts (HGF), suggesting a tumor specific cytotoxic activity. The cytotoxic activity of these extracts was enhanced by fractionation on silica gel [H2, A2, M1-M3] or ODS column chromatography [70M]. Several fractions [H2, H4, H5, A1, A2, A3, A5, A6, A7, M2] reversed the multidrug resistance (MDR) phenotype with L5178 mouse lymphoma T cells, more efficiently than (+/-)-verapamil. The extracts and fractions did not show any detectable anti-human immunodeficiency virus (HIV) or anti-Helicobacter pylori activity. Thus, this study suggests the effective and selective antitumor potential of 'Anastasia Red' of sweet pepper for further phytochemical and biological investigation. PMID- 12722140 TI - DNA degradation by water extract of green tea in the presence of copper ions: implications for anticancer properties. AB - In recent years a number of reports have documented the chemopreventive effect of green tea consumption on various types of cancers such as those of bladder, prostate, esophagus and stomach. This property is attributed to the presence in green tea of polyphenols known as catechins. These include epigallocatechin-3 gallate, epigallocatechin and epicatechin. In addition to their antioxidant properties plant derived polyphenolics are also capable of oxidative DNA damage particularly in the presence of transition metal ions. We have recently proposed a mechanism for cytotoxic action of plant-derived polyphenols against cancer cells that involves mobilization of endogenous copper and consequent prooxidant action. In partial support of the idea, in the present paper we show that water extract of green tea is considerably more efficient than black tea extract in DNA cleavage in the presence of copper ions. Green tea extract also shows a higher rate of Cu(II) reduction and consequent hydroxyl radical formation. Cu(II) reduction is presumably accompanied by the formation of 'oxidized species' of tea polyphenols, which in turn also appear to catalyze the reduction of Cu(II) leading to redox cycling of copper ions. The results are discussed in relation to the structural differences between polyphenols of green and black tea. PMID- 12722139 TI - Protective effects of Acanthopanax Radix extract against endotoxemia induced by lipopolysaccharide. AB - Endotoxemia causes an enhanced production of reactive oxygen radicals, which contribute to multiple organ dysfunction. When rats were given intravenous lipopolysaccharide and tested 6 h later we found that the activities of catalase and glutathione peroxidase (GSH-Px) in kidney, were acutely suppressed while in serum the levels of nitric oxide (NO), lipid peroxidation, urea nitrogen and creatinine were significantly increased, indicating the excessive production of reactive oxygen radicals and the presence of renal injury. Pretreatment of rats with Acanthopanax Radix extract administered orally for 30 days reduced the NO and lipid peroxidation levels, increased the activities of catalase and GSH-Px, and attenuated the renal dysfunction. These results suggested that scavenging of reactive oxygen radicals is part of the mechanism by which Acanthopanax Radix extract works as an effective agent in preventing multiple organ dysfunction. PMID- 12722141 TI - Acaricidal activity of Uvaria versicolor and Uvaria Klaineana (Annonaceae). AB - Three Uvaria species, namely U. klaineana, U. mocoli and U. versicolor were tested in vitro against Dermatophagoides pteronyssinus, the European house dust mite. The most active extracts were the Uvaria versicolor stems crude methanol and hexane extracts with EC(50) values of 0.095 g/m(2 )and 0.12 g/m(2), respectively. The bioassay-guided fractionation of the hexane extract led to the isolation of benzyl benzoate (1). which exhibited an EC(50) value of 0.045 g/m(2). A new fl avanone, versuvanone (2). and the known oxoaporphine liriodenine (3). were also isolated from this species and showed EC(50) values > 1.5 g/m(2). A weak acaricidal activity (0.85 g/m(2)) was observed for the dichloromethane extract of Uvaria klaineana, due to the presence of benzyl benzoate. Uvaria mocoli was inactive. The structures of compounds were elucidated using 2D-NMR techniques. PMID- 12722142 TI - Essential oil composition and antifungal activity of Foeniculum vulgare Mill obtained by different distillation conditions. AB - The influence of different hydrodistillation conditions was evaluated from the standpoint of essential oil yield, chemical composition and antifungal activity from seeds of Foeniculum vulgare Mill. Three hydrodistillation conditions were considered. The main constituents of the oils were: (E)-anethole (72.27%-74.18%), fenchone (11.32%-16.35%) and methyl chavicol (3.78%-5.29%). The method of distillation significantly effected the essential oil yield and quantitative composition, although the antifungal activity of the oils against some fungi was only slightly altered. PMID- 12722143 TI - Butanolides from Machilus thunbergii and their inhibitory activity on nitric oxide synthesis in activated macrophages. AB - In activated macrophages the inducible form of nitric oxide synthase (iNOS) generates high amounts of the toxic mediator, nitric oxide (NO), that contributes to the circulatory failure associated with septic shock. Three butanolides were isolated from Machilus thunbergii as active principles which inhibit the production of NO in lipopolysaccharide-activated RAW 264.7 cells, and their structures were identified as litsenolide A2 (1), B1 (2) and B2 (3). They showed dose-dependent inhibition of NO syntheses and the IC(50)s were 3.36, 3.70 and 6.19 micro m, respectively. These new inhibitors of iNOS may have potential in the treatment of endotoxaemia and inflammation accompanied by the overproduction of NO. PMID- 12722144 TI - In vitro antagonistic activity of monoterpenes and their mixtures against 'toe nail fungus' pathogens. AB - The antibiotic effect of the active ingredients in Meijer medicated chest rub (eucalyptus oil, camphor and menthol) as well as the inactive ingredients (thymol, oil of turpentine, oil of nutmeg and oil of cedar leaf) were studied in vitro using the fungal pathogens responsible for onychomycosis, such as the dermatophytes Tricophyton rubrum, Trichophyton mentagrophytes, Microsporum canis, Epidermophyton fl occosum and Epidermophyton stockdale. The zones of inhibition data revealed that camphor (1). menthol (2). thymol (3). and oil of Eucalyptus citriodora were the most efficacious components against the test organisms. The MIC(100) for mixtures of these four components in various carrier solvents revealed that formulations consisting of 5 mg/mL concentrations of each have a potential to be efffective in controlling onychomycosis. PMID- 12722145 TI - Antidiarrhoeal activity of hot water extract of black tea (Camellia sinensis). AB - The effect of a hot water extract of black tea (Camellia sinensis (L.) O. Kuntze, Theaceae) on upper gastrointestinal transit and on diarrhoea was investigated employing conventional rodent models of diarrhoea. Black tea extract was found to possess antidiarrhoeal activity in all the models of diarrhoea used. Naloxone (0.5 mg/kg i.p.) significantly inhibited the antidiarrhoeal activity of the extract as well as loperamide, thus indicating a role of the opioid system in the antidiarrhoeal activity of the extract. PMID- 12722146 TI - Crown gall -- a plant tumour with biological activities. AB - Petroleum ether, acetone, 80% MeOH and water extracts of crown gall, a plant tumour, obtained from Eucalyptus globulus tree were screened for cytotoxic, antioxidant, antiinflammatory, embryotoxic, antitumour-promoting and antimicrobial activities. In terms of bioactivity the 80% MeOH extract was most effective followed by the acetone extract. The petroleum ether extract showed weak to moderate cytotoxic activity in dose-dependent manner against PC12 cells, mouse L fibroblasts and 1321N1 glia cells, whereas the hydroalcohol extract had no or a weak cytotoxic effect. The 80% MeOH extract exhibited strong antioxidant activity. Based on the in vitro HET-CAM assay all the extracts were effective against inflammation. The extracts did not show any embryotoxic effect at the concentrations tested. Antitumour-promoting activity (100% inhibition; 100 microg/mL) was observed in the 80% MeOH and acetone extracts. In the antimicrobial screening all extracts displayed predominantly antifungal activity against Candida sp. The extracts also showed various levels of antibacterial activity against E. faecalis, Ps. aeruginosa, Bac. subtilis and Staph. epidermidis. From the results of the investigations it can be concluded that crown gall is a valuable plant tumour tissue having interesting biological activities. PMID- 12722147 TI - Duodenal ulcer prevalence: research into the nature of possible protective dietary lipids. AB - The varying geographical prevalence of duodenal ulceration has suggested a relationship to staple diet. Previous experiments on animal peptic ulcer models showed that certain foods, particularly the lipid fraction, are ulceroprotective. This paper reports experiments on animal models further to investigate the nature of the protective substances in the most active lipid, that of horse gram. The free fatty acids and triglycerides, sterols, sterol esters and phospholipids from horse gram were extracted and tested for protective activity on rat peptic ulcer models: the pyloric ligation model which is chronic, involving 14 days pre feeding, and two acute models using ethanol or cysteamine to induce ulceration. The results showed that sterol esters, but not sterols, were protective in the pyloric ligation model. Sterols were protective in the acute models using ethanol induced and cysteamine-induced ulceration. Phospholipids were protective in both types of model. The free fatty acids and triglycerides gave no protection using the pyloric ligation model. The presence of sterols, sterol esters and phospholipids in the lipid fraction of foods in staple diets may account for the low prevalence of duodenal ulcer in certain geographical areas, despite a uniformly high prevalence of Helicobacter pylori infection. PMID- 12722149 TI - Aphrodisiac activity of methanol extract of leaves of Passiflora incarnata Linn in mice. AB - The aphrodisiac properties of the methanol extract of leaves of Passiflora incarnata Linn. have been evaluated in mice by observing the mounting behaviour. The methanol extract of P. incarnata exhibited significant aphrodisiac behaviour in male mice at all doses, i.e. 75, 100 and 150 mg/kg. Amongst these, the highest activity was observed with the 100 mg/kg dose when the mountings were calculated about 95 min after the administration of the test extracts. PMID- 12722148 TI - Parmelia cirrhatum: a potential source of broad spectrum natural antifungal. AB - During antifungal screening of some plants, the water extract of lichen, Parmelia cirrhatum was found to be most effective against tested pathogenic fungi at the range of 60-80 microL/mL concentrations. The extract showed heavy doses of inoculum potential at 80 microL/mL and the antifungal activity of the extract did not expire up to 24 months of storage. The extract did not showed any irritation on mammalian skin up to 10% concentration. Thus, the water extract of Parmelia cirrhatum can be used a potential source of natural antifungal against fungal infections. PMID- 12722151 TI - Indonesian tea mistletoe (Scurrula oortiana) stem extract increases tumour cell sensitivity to tumour necrosis factor alpha (TNFalpha). AB - The water extract of Indonesian tea mistletoe, Scurrula oortiana ('benalu teh'), has been used for generations to treat tumours, however, little is known of its biological action. In the present study the stem and leaf extracts of S. oortiana were investigated for their modulation of tumour cell sensitivity toward TNFalpha, a potent anti-tumour cytokine. WEHI-164 cells which are known to be sensitive to TNFalpha, were used as a model. The assay results showed that either the stem or leaf extract of S. oortiana increased the sensitivity or susceptibility of WEHI-164 cells to TNFalpha as shown by decreases in LD(50) values in the TNFalpha sensitivity assay. The stem extract showed a greater increase than the leaf extract, increasing the sensitivity more than 160 times compared with the normal untreated tumour cells. This study showed for the fi rst time that water extracts of S. oortiana were significantly cytotoxic to the WEHI 164 tumour cell line and increased tumour cell sensitivity to TNFalpha mediated lysis. PMID- 12722150 TI - Effects of Zuccagnia punctata on the gastrointestinal tract in rats and mice. AB - A pharmacological evaluation of Zuccagnia punctata Cav. (Fabaceae) on the gastrointestinal tract was made in rats and mice. 2',4'-dihydroxychalcone and 2',4'-dihydroxy-3'-methoxychalcone were isolated from Zuccagnia punctata. The data reported in the present work indicate that the acetone extract and infusion of Zuccagnia punctata reduced intestinal transit in rats and mice and offered protection against ethanol-induced ulceration in rats. The Z. punctata effect could be due, in part, to the presence of 2',4'-dihydroxychalcone and 2',4' dihydroxy-3'-methoxychalcone in this plant. PMID- 12722152 TI - Hypoglycaemic and hypolipidaemic action of alcohol extract of Tinospora cordifolia roots in chemical induced diabetes in rats. AB - The present investigation was undertaken to evaluate the hypoglycaemic and hypolipidaemic effects of an alcohol extract of Tinospora cordifolia roots, an indigenous plant used in Ayurvedic medicine in India. Oral administration of the extract of Tinospora cordifolia (TCREt) roots for 6 weeks resulted in a significant reduction in blood and urine glucose and in lipids in serum and tissues in alloxan diabetic rats. The extract also prevented a decrease in body weight. Thus our study clearly shows that an alcohol TCREt has a hypoglycaemic and hypolipidaemic action. PMID- 12722153 TI - In vitro antimicrobial efficacy of leaf essential oils of Chukrasia tabularis Adr Juss and Melia dubia Cav (Meliaceae). AB - The essential oils of Chukrasia tabularis and Melia dubia were tested for their antimicrobial activity against ten different pathogenic microorganisms responsible for human pathologies using standard antimicrobial assays. Chukrasia tabularis leaf oil exhibited strong antibiotic activities against Proteus vulgaris and Fusarium oxysporum and did not show any activity against the tested bacteria. Melia dubia leaf essential oil exhibited bacteriostatic and fungistatic activities against Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae and Fusarium oxysporum and Candida albicans respectively. The inhibitory activities of both essential oils are comparable with that of respective standards. PMID- 12722154 TI - Secoiridoid glucosides with free radical scavenging activity from the leaves of Syringa dilatata. AB - Activity-guided fractionation of the EtOAc and MeOH extract of the leaves of Syringa dilatata NAKAI furnished one free radical scavenger, the secoiridoid glucoside oleuropein together with ligstroside and an iridoid glucoside, syringopicroside. Oleuropein interacted with the stable free radical, 1,1 diphenyl-2-picrylhydrazyl (DPPH), and showed an IC(50) value of 40.4 microM. L Ascorbic acid as a positive control showed an IC(50) value of 50.3 microM. PMID- 12722155 TI - Trypsin inhibitory effect of wedelolactone and demethylwedelolactone. AB - Wedelolactone (WL) and demethylwedelolactone (DWL) isolated from Eclipta alba were tested in the trypsin inhibition bioassay (in vitro). Both compounds showed potent activity. IC(50) values of WL and DWL were found to be 2.9 and 3.0 microg/mL respectively. PMID- 12722156 TI - Antinociceptive, antiinflammatory and acute toxicity effects of Salvia leriifolia Benth seed extract in mice and rats. AB - The antinociceptive and antiinflammatory effects as well as the acute toxicity of Salvia leriifolia aqueous seed extract were studied in mice and rats. Antinociceptive activity was assessed using the hot-plate and tail flick tests. The effect on acute inflammation was studied using vascular permeability increased by acetic acid and xylene-induced ear oedema in mice. The activity against chronic inflammation was assessed using the cotton pellet test in rats. The LD(50) of the extract was found to be 19.5 g/kg (i.p.) in mice. The aqueous seed extract showed significant and dose-dependent (1.25-10 g/kg) antinociceptive activity over 7 h, and was inhibited by naloxone pretreatment. Significant and dose-dependent (2.5-10 g/kg) activity was observed against acute inflammation induced by acetic acid and in the xylene ear oedema test. In the chronic inflammation test the extract (2.5-5 g/kg) showed significant and dose-dependent antiinflammatory activity. The aqueous seed extract of S. leriifolia may therefore have supraspinal antinociceptive effects which may be mediated by opioid receptors, and showed considerable effects against acute and chronic inflammation. PMID- 12722157 TI - Korean medicinal plants inhibiting to human immunodeficiency virus type 1 (HIV-1) fusion. AB - In order to find novel anti-HIV agents from natural products, 80 MeOH extracts of Korean plants were applied to a syncytia formation inhibition assay, which is based on the interaction between the HIV-1 envelope glycoprotein gp120/41 and the cellular membrane protein CD4 of T lymphocytes. The most potent HIV-1 fusion inhibition was shown by the stem bark of Ailanthus altissima with 74.9 +/- 4.4% at a concentration of 100 microg/mL. PMID- 12722158 TI - Cytoprotective activity of Amla (Emblica officinalis) against chromium (VI) induced oxidative injury in murine macrophages. AB - The cytoprotective and immunomodulating properties of Emblica officinalis (Amla) against chromium (VI) induced oxidative damage are reported. Chromium (VI) at 1 micro g/mL concentration was highly cytotoxic. It enhanced free radical production and decreased reduced glutathione (GSH) levels and glutathione peroxidase (GPx) activity in macrophages. The presence of Amla resulted in an enhanced cell survival, decreased free radical production and higher antioxidant levels similar to that of control cells. Further, chromium (VI) treatment resulted in decreased phagocytosis and gamma-interferon (gamma-IFN) production while Amla inhibited chromium induced immunosuppression and restored both phagocytosis and gamma-IFN production by macrophages significantly. PMID- 12722159 TI - Cryptolepine hydrochloride: a potent antimycobacterial alkaloid derived from Cryptolepis sanguinolenta. AB - The activity of cryptolepine hydrochloride, a salt of the main indoloquinoline alkaloid from the West African medicinal plant Cryptolepis sanguinolenta, was assessed against the fast growing mycobacterial species Mycobacterium fortuitum, which has recently been shown to be of use in the evaluation of antitubercular drugs. The low minimum inhibitory concentration (MIC) of this compound (16 microg/mL) prompted further evaluation against other fast growing mycobacteria namely, M. phlei, M. aurum, M. smegmatis, M. bovis BCG and M. abcessus and the MICs ranged over 2-32 microg/mL for these species. The strong activity of this agent, the need for new antibiotics with activity against Mycobacterium tuberculosis, coupled with the ethnobotanical use of C. sanguinolenta extracts to treat infections, highlight the potential of the cryptolepine template for development of antimycobacterial agents. PMID- 12722160 TI - Movement Disorders Society Scientific Issues Committee report: SIC Task Force appraisal of clinical diagnostic criteria for Parkinsonian disorders. AB - As there are no biological markers for the antemortem diagnosis of degenerative parkinsonian disorders, diagnosis currently relies upon the presence and progression of clinical features and confirmation depends on neuropathology. Clinicopathologic studies have shown significant false-positive and false negative rates for diagnosing these disorders, and misdiagnosis is especially common during the early stages of these diseases. It is important to establish a set of widely accepted diagnostic criteria for these disorders that may be applied and reproduced in a blinded fashion. This review summarizes the findings of the SIC Task Force for the study of diagnostic criteria for parkinsonian disorders in the areas of Parkinson's disease, dementia with Lewy bodies, progressive supranuclear palsy, multiple system atrophy, and corticobasal degeneration. In each of these areas, diagnosis continues to rest on clinical findings and the judicious use of ancillary studies. PMID- 12722161 TI - Effect of pulsatile administration of levodopa on dyskinesia induction in drug naive MPTP-treated common marmosets: effect of dose, frequency of administration, and brain exposure. AB - Levodopa (L-dopa) consistently primes basal ganglia for the appearance of dyskinesia in parkinsonian patients and 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine hydrochloride (MPTP) -treated primates. This finding may reflect its relatively short duration of effects resulting in pulsatile stimulation of postsynaptic dopamine receptors in the striatum. We have compared the relationship between L-dopa dose and frequency of administration on dyskinesia initiation in drug-naive, MPTP-treated common marmosets. We have also studied the effect of increased brain exposure to pulsatile administration by combining a low-dose of L-dopa with the peripheral catechol-O-methyltransferase inhibitor (COMT-I), entacapone. Pulsatile administration of a low (dose range, 5.0-7.5 mg/kg p.o.) or a high (12.5 mg/kg) dose of L-dopa plus carbidopa b.i.d. produced a dose-related reversal of motor deficits. Repeated administration of low and high doses of L-dopa for 26 days to drug-naive, MPTP-treated animals also caused a dose-related induction of peak-dose dyskinesia. Repeated administration of high-dose L-dopa b.i.d. compared to once daily caused a frequency-related improvement of motor symptoms, resulting in a more rapid and initially more intense appearance of peak-dose dyskinesia. Administration of low-dose L-dopa b.i.d. for 26 days in combination with entacapone enhanced the increase in locomotor activity and reversal of disability produced by L-dopa alone, but with no obvious change in duration of L-dopa's effect. However, combining entacapone with L-dopa resulted in the more rapid appearance of dyskinesia, which was initially more severe than occurred with L-dopa alone. Importantly, increasing pulsatile exposure of brain to L-dopa by preventing its peripheral breakdown also increases dyskinesia induction. PMID- 12722162 TI - Fear of falling and postural control in Parkinson's disease. AB - This study investigated the relationship between fear of falling (FOF) and qualitative and quantitative postural control in Parkinson's disease (PD). Fifty eight nondemented PD patients were studied along with age-matched healthy controls. The degree of FOF was estimated using the Activities-specific Balance Confidence scale. Qualitative postural control was evaluated using a component of the Unified Parkinson Disease Rating Scale. Postural control was quantified, using centre of pressure measures obtained from a force plate, for eight standing balance tests of different challenges. The results showed that FOF was more evident for PD patients when compared with healthy individuals of similar age. Furthermore, FOF was significantly associated with a qualitative estimate of postural control in PD; individuals with PD who had a greater degree of posture impairment reported greater FOF. The results also showed that an estimate of FOF may help to explain quantitative postural instability in PD. FOF, when coupled with a qualitative estimate of postural control, was able to explain a greater amount of variation in quantitative balance performance for five of the eight balance tests. When considered independently, the qualitative measure of postural control, in general, could not well predict quantitative balance performance. The greater degree of FOF and its possible association with altered postural control suggests that FOF should be considered as an important, independent risk factor in the assessment and treatment of postural instability in patients with PD. PMID- 12722163 TI - Psychosocial problems in Parkinson's disease: evaluation of a disease-specific questionnaire. AB - Our objective was to evaluate the BELA-P-k, a questionnaire for measuring psychosocial problems and need for help in Parkinson's disease (PD) patients. The Belastungsfragebogen Parkinson kurzversion (BELA-P-k) was translated from German into Dutch. It consists of 19 items distributed over four subscales: achievement capability/physical symptoms, fear/emotional functioning, social functioning and partner-bonding/family, with a "Bothered by" (Bb) and a "Need for Help" (NfH) score. The BELA-P-k was tested for cultural differences, relevance, and feasibility in a pilot study (n = 10) and compared in a validation study (n = 54) with the Sickness Impact Profile, the COOP/WONCA Functional Health Assessment Charts and the Loneliness Questionnaire. All questionnaires were administered in person at home, in a prescribed order. The BELA-P-k was completed by 64 patients with PD. The internal-consistency reliability coefficients for the total Bb (0.90) and NfH (0.93) scales were excellent. The internal consistency of the subscales exceeded the 0.70 standard except for the "Bothered by partner bonding/family scale" (0.61). Almost all BELA-P-k subscales correlated highly (P < 0.001) with the corresponding scales of the standard quality-of-life indices. There was no significant relationship between disease severity (Hoehn and Yahr) and the BELA-P-k. We conclude that the BELA-P-k is a relevant, reliable and valid measure for assessing psychosocial problems and need for help of PD patients. PMID- 12722164 TI - Long-term outcome of quetiapine use for psychosis among Parkinsonian patients. AB - To evaluate the long-term efficacy and tolerability of quetiapine for psychosis among parkinsonian patients, a retrospective analysis of all parkinsonian patients taking quetiapine for psychosis in a single movement disorders center was carried out. Demographic data, including type and severity of psychosis, presence of dementia, treatment response, before and after Unified Parkinson's Disease Rating Scale (UPDRS)-motor scores and Hoehn and Yahr (H&Y) scale were obtained. One hundred six parkinsonian patients with a mean age of 76.6 years were on an average levodopa (L-dopa) dose of 415 mg/d. Seventy-eight of 106 (74%) remained on quetiapine for a mean duration of 15 months at an average dose of 60 mg per day. Eighty-seven (82%) patients had partial or complete resolution of their psychosis whereas 19 (18%) patients had no improvement on quetiapine. Motor worsening was noted in 34 (32%) patients but was uncommonly sufficient to warrant quetiapine discontinuation. More quetiapine non-responders were noted to be demented, delusional, and experienced threatening psychosis but only the presence of dementia remained significant on multivariate analysis (OR = 11.6; 95% CI = 1.4-92.9). Also, patients who developed motor worsening while on quetiapine tended to be more demented (P = 0.07). PMID- 12722165 TI - Effects of unilateral pallidotomy on voluntary movement, and simple and choice reaction times in Parkinson's disease. AB - We studied the effects of unilateral pallidotomy on motor execution and reaction times in patients with moderately advanced Parkinson's disease (PD). Twelve consecutive patients (7 men, 5 women; all right-handed) underwent left-side microelectrode-guided pallidotomy. In addition to clinical rating, reaction time (RT) tests and repetitive movements of the contralesional hand/arm were carried out at baseline and 2 to 3 months after surgery while patients were on optimal medical regimens (on period). The initiation time in both simple reaction time (SRT) and choice reaction time (CRT) improved significantly after pallidotomy (P < 0.05), whereas no effect was observed on the choice processing time, which was calculated by subtracting the mean value of the onset of SRT from that of CRT. Pallidotomy resulted in significant improvement of repetitive movements such as hand pronation/supination and finger-tapping (P < 0.002, P < 0.005, respectively). Improvements in RT tests and repetitive movements suggest that pallidotomy may enhance attention and motor function. These effects are probably mediated through the pallido-thalamic-cortical neural circuitry. PMID- 12722166 TI - Acute psychotropic effects of bilateral subthalamic nucleus stimulation and levodopa in Parkinson's disease. AB - High-frequency deep brain stimulation (DBS) of the subthalamic nucleus (STN) improves the motor symptoms of Parkinson's disease (PD). Opposite changes in mood, such as mania or depression, have been reported after surgery, but it is not known whether these side effects are specifically related to STN DBS. To learn whether STN DBS also influences the limbic loop, we investigated acute subjective psychotropic effects related to levodopa or bilateral STN DBS. After a median postoperative follow-up of 12 months, 50 PD patients completed the Addiction Research Center Inventory (ARCI), assessing subjective psychotropic effects in four conditions: off-drug/on-stimulation; off-drug/off-stimulation; on drug/off-stimulation; and on-drug/on-stimulation. Both levodopa and STN DBS improved all the ARCI subscales, indicating subjective feelings of well being, euphoria, increase in motivation, and decrease in fatigue, anxiety, and tension. A suprathreshold dose of levodopa was significantly more effective than STN DBS, using the same electrical parameters as for chronic stimulation, on four of the five ARCI subscales. We concluded that 1) both STN DBS and levodopa have synergistic acute beneficial psychotropic effects in PD, 2) the psychotropic effects of both treatments need to be considered in the long-term management of chronic STN DBS, and 3) the results indicate an involvement of the limbic STN in mood disorders of PD. PMID- 12722167 TI - Treatment of advanced Parkinson's disease by subthalamotomy: one-year results. AB - We studied effects on parkinsonian features at 6 and 12 months in 12 patients who underwent unilateral ablation of subthalamic nucleus (STN). Microelectrode mapping was used, and a lesion was created in the STN using thermal coagulation and confirmed with magnetic resonance imaging. At 6 months postoperatively, improvements were seen in several areas: 1) Unified Parkinson's Disease Rating Scales II and III (UPDRS II and III) scores, 30% in off period, 38% in on period; 2) Schwab and England Scale (S&E) score, 21%; and 3) on dyskinesia, 85%. Contralateral rigidity, bradykinesia, UPDRS II and III scores, and S&E scores remained improved at 12 months. Daily dosage of levodopa requirement was reduced by 42%. Axial motor features, gait, postural stability, off period tremor, and motor fluctuation improved at 6 and 12 months but showed a decline in benefits at 18 months. Complications include 3 cases of hemiballism, of whom 2 patients recovered spontaneously but 1 died from aspiration pneumonia. One patient had asymptomatic hematoma, and 2 suffered transient postural asymmetries. We conclude that unilateral subthalamotomy results in moderate improvement in all aspects of parkinsonian features, allows reduction in the dose of levodopa required, and ameliorates drug-induced complications throughout 12-month assessments. PMID- 12722168 TI - Executive cognitive deficits in primary dystonia. AB - Primary dystonia is a disorder of movement for which no consistent pathophysiology has been identified; in the absence of evidence to the contrary, it is assumed to be cognitively benign. We have studied a clinically heterogeneous group of 14 patients with primary dystonia on a battery of neuropsychological tests. Despite well-preserved speed of information processing, language, spatial, memory and general intellectual skills relative to normal controls, we have identified a constellation of attentional-executive cognitive deficits on the Cambridge Neuropsychological Test Automated Battery (CANTAB). Specifically, patients demonstrated significant difficulties negotiating the extra-dimensional set-shifting phase of the IED task. The implications of these findings for the pathophysiology of primary dystonia are discussed. This is, to the best of our knowledge, the first report of a significant cognitive deficit in patients with primary dystonia. PMID- 12722169 TI - Alternating leg muscle activation during sleep and arousals: a new sleep-related motor phenomenon? AB - We describe a quickly alternating pattern of anterior tibialis activation, recorded during nocturnal polysomnography in 16 patients. Polysomnography, usually for sleep-disordered breathing, included surface electromyograms over the anterior tibialis of each leg. Cases were identified from approximately 1,500 studies reviewed in the course of standard clinical care. Patients were 12 men and 4 women (mean age, 41 +/- 15 years; range, 12-70 years). Brief activation of the anterior tibialis in one leg alternated with similar activation in the other leg. Activations occurred at a frequency of approximately 1 to 2 Hz, each lasted between 0.1 and 0.5 seconds, and sequences of alternating activations usually lasted between several and 20 seconds. The phenomenon occurred in all sleep stages but particularly during arousals. Ten of the 16 patients had periodic leg movements during sleep at a rate >/= 5.0 per hour, and 12 of the 16 patients were taking antidepressant medication. Alternating leg muscle activation (ALMA) during sleep, at this relatively high frequency, may be a newly described phenomenon. We speculate that ALMA could represent transient facilitation of a spinal central pattern generator for locomotion, perhaps due to serotonergic effects of antidepressant medication. PMID- 12722170 TI - Impairment of individual finger movements in Parkinson's disease. AB - By analyzing the kinematics of repetitive, constant-amplitude, finger oppositions, we compared the impairment of individual and nonindividual finger movements in patients with Parkinson's disease. In one task, subjects tapped only the index finger against the thumb (individual oppositions); in the other task, they tapped all four fingers together against the thumb pad (nonindividual oppositions). We used an optoelectronic motion analysis system to record movements in three-dimensional space and recorded three 5-second trials for each task. We counted how many finger oppositions subjects performed during each trial and measured the duration and amplitude of the flexions and extensions. We also calculated the duration of the pauses after flexion and extension. We assessed the deterioration of motor performance in patients by investigating the changes in speed and amplitude with task completion. During both tasks, normal subjects and patients performed finger flexions faster than extensions, and they invariably paused longer after flexion than after extension. Patients performed individual and nonindividual finger movements slowly and with reduced amplitude. Patients were disproportionately slow during flexion and in switching from flexion to extension. Movement slowness increased as finger oppositions progressed but predominantly when patients had to move fingers individually. In conclusion, in patients with Parkinson's disease, the motor performance deteriorated with task completion more during individual than during nonindividual finger movements. Parkinson's disease, therefore, impairs individual finger movements more than gross hand movements. This distinction reflects the finer cortical control needed to promote and sustain this highly fractionated type of motor output. PMID- 12722171 TI - Manual transport in Parkinson's disease. AB - We analyze hand dexterity in Parkinson's disease patients (PD) and control subjects using a natural manual transport task (moving an object from one place to another). Eight PD patients and 10 control subjects carried out the task repeatedly at maximum speed both in off and on medicated status. The movement parameters and the grip and load forces were recorded. Using the force and velocity signals, 10 subsequent phases of the transport movement were defined and their durations were measured. The difference between the control group and the test group in off and on was established statistically using non-parametric methods. There was slowed reaching and a striking disturbance of establishing the precision grip in PD. The transport capabilities were impaired differentially. Although acceleration and reaching sufficient height of the lift were disturbed in PD subjects, transport of the object toward the target position was almost normal. A partial disturbance was observed when cancelling the grip. Dopaminergic medication improved only specific hand skills, especially establishment of the precision grip and one of the four transport phases. A long movement path was more sensitive for movement disturbance in Parkinson's disease than a short one. PMID- 12722173 TI - Clinical and economic analysis of spa therapy in Parkinson's disease. AB - The effectiveness of spa therapy in the management of patients with Parkinson's disease (PD) has never been evaluated. This is assessed in this pilot study. A prospective, randomized, cross-over, controlled study was conducted in 31 PD patients who underwent a 20-week spa period, including spa therapy for 3 weeks, and a 20-week non-spa period. Effectiveness was assessed using quality of life scales (PDQ-39 and SF-36), motor scale (UPDRS) and psychological questionnaire (GHQ-28), at baseline and at 4 (T4) and at 20 weeks (T20). Direct medical costs (radiological and laboratory tests, physician fees, drug therapy, and ancillary care) were recorded over each 20-week period. At T4, spa therapy improved significantly several dimensions of PDQ-39 and SF-36, part IV of the UPDRS, and GHQ-28. At T20, no difference in any parameter was found. The mean direct medical cost over 20 weeks (euro;1,328 +/- 167; pound 776 +/- 97 per patient) in the spa period was slightly but significantly reduced in comparison with that of the non spa period (euro;1380 +/- 523; pound 807 +/- 306 per patient). This cost effectiveness analysis suggests that spa therapy is more effective and less expensive than conventional treatment alone and could be beneficial in the management of PD. PMID- 12722172 TI - Short-term effect of a single levodopa dose on micturition disturbance in Parkinson's disease patients with the wearing-off phenomenon. AB - We investigated the short-term effects of a single dose of levodopa (L-dopa) on micturition function in PD patients with wearing-off phenomenon. Eighteen PD patients who had median Hoehn and Yahr scores of 5 during the off phase and 3 during the on phase were recruited. We carried out urodynamic studies before and about 1 hour after the patients had taken 100 mg of L-dopa with dopa decarboxylase inhibitor (DCI). After taking the L-dopa/DCI, urinary urgency and urge incontinence aggravated, whereas voiding difficulty was alleviated in all 12 patients. When compared to the baseline assessment, urodynamic study results after taking 100 mg of L-dopa/DCI showed aggravated detrusor hyperreflexia; decreased maximum bladder capacity (P = 0.006); an increased maximum Watts Factor value (P = 0.001), reflecting the detrusor power on voiding; an increased Abrams Griffiths number (P = 0.042), reflecting urethral obstruction on voiding; decreased residual urine volume (P = 0.025); and increased static urethral closure pressure (P = 0.012). One hundred milligrams of L-dopa/DCI worsened detrusor hyperreflexia, producing worsened urinary urgency and urge incontinence during the storage (bladder-filling) phase. It also increased detrusor contractility much more than it did urethral obstruction in the voiding phase, producing overall lessening of voiding difficulty and improving voiding efficiency in our PD patients with the wearing-off phenomenon. PMID- 12722174 TI - Mirtazapine in essential tremor: a double-blind, placebo-controlled pilot study. AB - We sought to determine whether mirtazapine is safe and well-tolerated as a treatment for essential tremor (ET). We studied mirtazapine in a randomized, double-blind, placebo-controlled, crossover study of 17 ET patients. Patients were started with 15 mg per day of either mirtazapine or placebo for 1 week and the dose was escalated weekly until the targeted dose of 45 mg per day was achieved. This dose was maintained for 2 weeks. Tremor was assessed at baseline and after 14 days of 45 mg of mirtazapine or placebo. There was a minimum washout period of 14 days between the two arms of the study. Tremor assessments included global improvement, Fahn Tolosa Marin Tremor Rating Scale, Beck Depression Inventory and the Parkinson's Disease Questionnaire-39. Patient global improvement ratings indicated that in the placebo condition 12 patients were unchanged and 1 patient was mildly improved. In the mirtazapine condition, 10 patients were unchanged, 2 were moderately improved and 1 was markedly improved. There was no significant improvement with mirtazapine or placebo compared to baseline as measured by the Tremor Rating Scale. Adverse effects were more common in the mirtazapine group and included drowsiness, confusion, dry mouth, weight gain, polyuria, itching, nausea, gait and balance problems, blurred vision, and bad taste. We conclude that the majority of the ET patients do not benefit from mirtazapine. Mirtazapine has significant adverse effects and should be used cautiously in ET patients. PMID- 12722175 TI - Trigemino-facial inhibitory reflexes in idiopathic hemifacial spasm. AB - We investigated trigemino-facial excitatory and inhibitory responses in perioral muscles in hemifacial spasm (HFS). We examined 15 patients affected with idiopathic HFS and 8 healthy controls. Five patients had spasms mostly limited to the periocular region and 10 had spasms also involving the perioral muscles. Responses were recorded from the resting orbicularis oculi (OOc), levator labii superioris (LLS) and orbicularis oris (OOr) muscles, after supraorbital (SO) nerve stimulation and during isolated voluntary contraction of LLS muscle. Eight patients showed complete or partial preservation of the late silent period (SP2) in activated LLS muscle. The remaining 7 patients showed absence of SP2. Early and late excitatory responses were variably present in LLS muscle at rest. Patients with HFS clinically restricted to periocular muscles had at least partial preservation of the SP2. In conclusion, in HFS patients inhibitory trigemino-facial reflexes are impaired and excitatory trigemino-facial responses are elicited in perioral muscles. These two phenomena seem to develop independently; the degree of trigemino-facial reflex impairment parallels the extension of involuntary movements to the lower facial muscles. PMID- 12722177 TI - Phenotypic presentation of frontotemporal dementia with Parkinsonism-chromosome 17 type P301S in a patient of Jewish-Algerian origin. AB - A 39-year-old old Jewish woman of Algerian origin developed a rapidly progressive neurocognitive disorder characterized by asymmetric rigidity, spasticity with bilateral Babinski's sign, bradykinesia, altered speech that progressed to mutism, and severe bradyphrenia. She partially responded to levodopa. The family history revealed 4 affected first-degree relatives (3 had already died). Genetic studies carried out in the proband and her living affected sister showed a P301S mutation in chromosome 17. PMID- 12722176 TI - Dopamine D2 receptor TaqIA and TaqIB polymorphisms in Parkinson's disease. AB - In a case control study, we examined the association of DRD2 Taq1A and Taq1B polymorphisms and risk of PD, and evaluated the strength of linkage disequilibrium of the polymorphisms. The Taq1A and Taq1B polymorphisms were in strong linkage disequilibrium. There was, however, no significant association of the two polymorphisms with PD. PMID- 12722179 TI - Digital video. PMID- 12722178 TI - Pseudoathetosis in a patient with leprosy. AB - A 35-year-old man with borderline tuberculoid leprosy developed Type I lepra reaction 12 days after anti-leprosy treatment. There was acute worsening of neuropathic symptoms and skin lesions. He developed severe sensory ataxia and pseudoathetosis resulting in marked disability. His symptoms significantly improved on corticosteroid therapy. PMID- 12722180 TI - The Gts1 protein stabilizes the autonomous oscillator in yeast. AB - Continuous cultures of Saccharomyces cerevisiae show a robust autonomous temperature compensated oscillation in many metabolic functions. Respiratory activity, a convenient output to measure, oscillates with a period of 40 min. Deletion of GTS1, whose protein product has homology to the circadian per protein, has been implicated in temporal events within yeast, causes a reduction in periodicity to 18 min (wild-type period 40-60 min). The dilution rate was steadily increased from 0.04/h to 0.085/h and the oscillation stabilized after four to six dilutions. However, Gts1p's involvement in the maintenance and generation of metabolic synchrony, and in the central oscillating loop, appear to be minimal, as the mutant oscillation was robust and autonomous. Deletion of GTS1 did cause decreased temperature compensation of the period of the oscillation from Q(10) = 1.07 for the wild-type to Q(10) = 1.6 for the mutant. Also the degree of nutrient compensation observed for the wild-type was not observed in the GTS1-null mutant strain. It is postulated that Gts1p is involved in the mechanism that communicates external conditions, such as temperature, to the central oscillating loop. PMID- 12722181 TI - cDNA cloning, biochemical and phylogenetic characterization of beta- and beta' subunits of Candida albicans protein kinase CK2. AB - We have previously reported that Candida albicans protein kinase CK2 is composed of two distinct catalytic (alpha- and alpha'-) and two distinct regulatory (beta- and beta'-) subunits. We report here the isolation of two cDNAs clones, CaCKB1 and CaCKB2, encoding C. albicans beta- and beta'-subunits, respectively. The predicted beta- and beta'-proteins have calculated molecular masses of 34 kDa and 31 kDa and show all major features conserved in beta-subunits of other organisms, including the N-terminal autophosphorylation site, the internal acidic region and a potential metal-binding motif. The deduced amino acid sequence of C. albicans beta-subunit displays 48% identity with that of Saccharomyces cerevisiae and has an unusually long C-terminal acidic region containing a putative autophosphorylation site. C. albicans beta' shows 54% sequence identity with its S. cerevisiae homologue. Semi-quantitative RT-PCR analyses indicate that the mRNAs corresponding to both subunits are present in similar amounts in the yeast and hyphal forms of the fungus. To evaluate the biochemical properties of C. albicans beta- and beta'-subunits, both proteins were expressed in Escherichia coli and purified. Experiments performed in vitro indicate that both recombinant subunits reconstitute a fully functional holoenzyme when incubated with stoichiometric amounts of human recombinant alpha-subunit, as judged by their ability to abolish basal phosphorylation of calmodulin by human recombinant alpha subunit and the reversion of the inhibitory effect by polylysine. In addition, both regulatory subunits can be phosphorylated by human recombinant alpha subunit. Phylogenetic analysis of beta- and beta'-proteins of C. albicans and other organisms shows that the CKB gene duplication occurred before the split of the ascomycete and basidiomycete lineages. PMID- 12722182 TI - Cu,Zn superoxide dismutase in Rhodotorula and Udeniomyces spp. isolated from sea water: cloning and sequencing the encoding region. AB - The gene encoding the copper-zinc superoxide dismutase enzyme (SODC or Cu,Zn-SOD) has been cloned from several species of higher eukaryotes, but superoxide dismutase genes from moulds and yeast have not been studied extensively. Only 15 nucleotide sequences have been reported in the SwissProt, EMBL and GenBank data libraries. In general the presence of Cu,Zn-SOD in cytosol, as well as Mn-SOD in the mitochondrial matrix of yeast, has been accepted. The absence of Cu,Zn-SOD in a pigmented yeast has been accepted as a general rule. Some authors suggest that the absence of Cu,Zn-SOD in pigmented yeast is complemented by the presence of carotenoproteins that act as an extra mitochondrial antioxidant. In this report, we found that the absence of SODC is not a rule for pigmented yeast: Udeniomyces puniceus expresses an active SODC which responds to Cu(2+) induction, as has been reported previously for non-pigmented yeast. The encoding region of the sod1 gene was cloned from three species of pigmented marine yeast thorough genomic DNA PCR amplification. Fragments of 485-487 nucleotides were obtained, which contain information for theoretical products of 153-154 amino acids. In Rhodotorula mucilaginosa the deduced amino acid sequence shows that insertion of three bases (C(112), A(149) and C(166)) generates a stop codon at position 123 (TGA). For Rhodotorula graminis a single change (T for A) generates a stop codon at position 298. For both species, this non-transcription of encoding sequence correlates with the absence of peptides or active proteins in cell homogenates. For U. puniceus, the cloned nucleotide sequence contains all necessary information to produce a functional protein, which correlates with activity detected in cell homogenates, both under normal conditions and by copper induction experiments. Finally, we clearly showed that the key factor in protection against oxidative stress on pigmented yeast is related not only to the presence of protective pigments but also to their amounts and spectra, as well as the presence and activity of SODC. PMID- 12722184 TI - A history of research on yeasts 5: the fermentation pathway. PMID- 12722183 TI - Ty3/gypsy-like retrotransposons in Candida albicans and Candida dubliniensis: Tca3 and Tcd3. AB - Ty3/gypsy retrotransposons are a widespread group of eukaryote mobile genetic elements. They are similar in structure to, and may be ancestors of, the vertebrate retroviruses. Here we describe the first Ty3/gypsy retrotransposons from the pathogenic yeasts Candida albicans and Candida dubliniensis, which we refer to as Tca3 and Tcd3, respectively. Tca3 was first identified in a variety of strains as an element lacking a large part of its coding region. Comparative analyses between C. albicans and C. dubliniensis allowed us to identify the closely related full-length Tcd3 element, and, subsequently, the full-length Tca3 elements. The full-length versions of Tca3 and Tcd3 are broadly similar in structure to other Ty3/gypsy elements, but have several features of special interest, e.g. both elements appear to have a novel mechanism for priming minus strand DNA synthesis, probably involving conserved secondary structures adjacent to the 5' LTRs. Also, while closely related to each other, the two elements appear to be fairly distantly related to other known Ty3/gypsy-like elements. Finally, the occurrence of the internally deleted forms of Tca3 in many strains raises interesting questions concerning the evolution of these transposable elements in Candida and the evolution of Candida itself. The sequences reported in this paper have been assigned GenBank Accession Nos AF499463, AF499464 and AF510498. PMID- 12722185 TI - Associating protein activities with their genes: rapid identification of a gene encoding a methylglyoxal reductase in the yeast Saccharomyces cerevisiae. AB - Methylglyoxal is associated with a broad spectrum of biological effects, including cytostatic and cytotoxic activities. It is detoxified by the glyoxylase system or by its reduction to lactaldehyde by methylglyoxal reductase. We show that methylglyoxal reductase (NADPH-dependent) is encoded by GRE2 (YOL151w). We associated this activity with its gene by partially purifying the enzyme and identifying by MALDI-TOF the proteins in candidate bands on SDS-PAGE gels whose relative intensities correlated with specific activity through three purification steps. The candidate proteins were then purified using a glutathione-S transferase tag that was fused to them, and tested for methylglyoxal reductase activity. The advantage of this approach is that only modest protein purification is required. Our approach should be useful for identifying many of the genes that encode the metabolic pathway enzymes that have not been associated with a gene (about 275 in S. cerevisiae, by our estimate). PMID- 12722186 TI - [Infectious diseases in the global days. Present situation in Japan--infectious disease surveillance]. PMID- 12722187 TI - [Infectious diseases in the world--global health threat]. PMID- 12722188 TI - [Strategies for the prevention and control of infectious diseases in Japan]. PMID- 12722189 TI - [Medical practitioner's role on the communicable diseases control law]. PMID- 12722190 TI - [Changes in bacterial infectious diseases]. PMID- 12722191 TI - [History of deep-seated mycoses]. PMID- 12722192 TI - [Changes of parasitic and protozoan diseases in Japan]. PMID- 12722193 TI - [Nosocomial infections surveillance in Japan]. PMID- 12722194 TI - [Germs and toxins in bioterrorism]. PMID- 12722195 TI - [Japanese encephalitis]. PMID- 12722196 TI - [Influenzal encephalopathy]. PMID- 12722197 TI - [Herpes simplex virus encephalitis]. PMID- 12722198 TI - [Aseptic meningitis]. PMID- 12722199 TI - [Poliomyelitis]. PMID- 12722200 TI - [JC virus infection (progressive multifocal leukoencephalopathy)]. PMID- 12722201 TI - [Borna disease virus infection]. PMID- 12722203 TI - [Common cold (rhinovirus)]. PMID- 12722204 TI - [Viral upper respiratory tract infection]. PMID- 12722202 TI - [Influenza]. PMID- 12722205 TI - [Bronchiolitis (RS virus infection)]. PMID- 12722206 TI - [Rotavirus gastroenteritis]. PMID- 12722207 TI - [Norwalk virus infection]. PMID- 12722208 TI - [Astrovirus and enteric adenovirus]. PMID- 12722210 TI - [Erythema infectiosum]. PMID- 12722209 TI - [Rubella and congenital rubella syndrome]. PMID- 12722211 TI - [Exanthem subitum (roseola infantum)]. PMID- 12722212 TI - [Hand, foot and mouth disease]. PMID- 12722213 TI - [Herpes simplex virus infection]. PMID- 12722214 TI - [Varicella-zoster virus infection]. PMID- 12722215 TI - [Molluscum contagiosum]. PMID- 12722216 TI - [Adult T-cell leukemia]. PMID- 12722217 TI - [Kaposi's sarcoma]. PMID- 12722218 TI - [Epstein-Barr virus-associated diseases--from infectious mononucleosis to malignant neoplasms]. PMID- 12722219 TI - [Hepatitis A]. PMID- 12722220 TI - [Hepatitis B]. PMID- 12722222 TI - [Hepatitis D virus infection]. PMID- 12722221 TI - [Hepatitis C]. PMID- 12722223 TI - [Hepatitis E]. PMID- 12722224 TI - [Infection of TT virus]. PMID- 12722225 TI - [Cytomegalovirus infection]. PMID- 12722226 TI - [Adenovirus infection]. PMID- 12722227 TI - [MUMPS]. PMID- 12722228 TI - [AIDS]. PMID- 12722229 TI - [Viral hemorrhagic fever]. PMID- 12722230 TI - [West Nile fever/encephalitis]. PMID- 12722231 TI - [Nipah virus infection]. PMID- 12722233 TI - [Dengue/dengue hemorrhagic fever]. PMID- 12722232 TI - [Rabies]. PMID- 12722234 TI - [Hantavirus pulmonary syndrome]. PMID- 12722235 TI - [Staphylococcal infections]. PMID- 12722236 TI - [Hemolytic streptococcal infections]. PMID- 12722237 TI - [Streptococcus pneumoniae infection]. PMID- 12722238 TI - [Enterococcus infection]. PMID- 12722239 TI - [Meningococcal disease]. PMID- 12722240 TI - [Gonococcal infection]. PMID- 12722241 TI - [Moraxella (Branhamella) catarrhalis infection]. PMID- 12722242 TI - [Listeriosis]. PMID- 12722243 TI - [Anthrax]. PMID- 12722244 TI - [Nocardiosis]. PMID- 12722245 TI - [Actinomycosis]. PMID- 12722246 TI - [Escherichia coli infections]. PMID- 12722247 TI - [Klebsiella infection]. PMID- 12722249 TI - [Enterobacter, Citrobacter, Proteus]. PMID- 12722248 TI - [Serratia infections]. PMID- 12722250 TI - [Typhoid and paratyphoid fever]. PMID- 12722251 TI - [Salmonella infections]. PMID- 12722252 TI - [Shigellosis]. PMID- 12722253 TI - [Campylobacter infection]. PMID- 12722254 TI - [Haemophilus influenzae infection]. PMID- 12722255 TI - [Pseudomonas aeruginosa infection]. PMID- 12722256 TI - [Melioidosis]. PMID- 12722257 TI - [Pertussis]. PMID- 12722258 TI - [Legionnaires' disease]. PMID- 12722259 TI - [Helicobacter pylori infection]. PMID- 12722260 TI - [Brucellosis]. PMID- 12722261 TI - [Plague]. PMID- 12722262 TI - [Yersiniosis (except the plague)]. PMID- 12722264 TI - [Tetanus]. PMID- 12722263 TI - [Cat scratch disease]. PMID- 12722266 TI - [Anaerobic abscess]. PMID- 12722265 TI - [Pseudo-membranous colitis]. PMID- 12722267 TI - [Chlamydia trachomatis]. PMID- 12722268 TI - [Chlamydia (Chlamydophila) pneumoniae pneumonia]. PMID- 12722269 TI - [Epidemic typhus]. PMID- 12722270 TI - [Tsutsugamushi disease]. PMID- 12722271 TI - [Q fever]. PMID- 12722272 TI - [Ehrlichiosis]. PMID- 12722273 TI - [Tuberculosis]. PMID- 12722275 TI - [Leprosy]. PMID- 12722274 TI - [Nontuberculous mycobacteriosis]. PMID- 12722276 TI - [Mycoplasma pneumoniae pneumonia]. PMID- 12722277 TI - [Syphilis]. PMID- 12722278 TI - [Relapsing fever]. PMID- 12722279 TI - [Lyme disease]. PMID- 12722280 TI - [Leptospirosis]. PMID- 12722283 TI - [Pneumocystis carinii pneumonia]. PMID- 12722282 TI - [Aspergillosis]. PMID- 12722281 TI - [Candidiasis]. PMID- 12722284 TI - [Coccidioidomycosis]. PMID- 12722285 TI - [Protozoan diseases--overview]. PMID- 12722286 TI - [Amebic dysentery]. PMID- 12722287 TI - [Malaria]. PMID- 12722288 TI - [Toxoplasmosis]. PMID- 12722289 TI - [Cryptosporidiosis]. PMID- 12722291 TI - [Giardiasis]. PMID- 12722290 TI - [Cyclosporiasis]. PMID- 12722292 TI - [Babesiosis]. PMID- 12722293 TI - [Parasitic infections--introduction]. PMID- 12722294 TI - [Echinococcosis]. PMID- 12722295 TI - [Strongyloidiasis]. PMID- 12722296 TI - [Scabies]. PMID- 12722297 TI - [Guidelines for the diagnosis and treatment of hemolytic uremic syndrome (HUS) with enterohemorrhagic colitis (Japanese Society of Pediatric Nephrology)]. PMID- 12722298 TI - [Guideline for the treatment of Hansen's disease]. PMID- 12722299 TI - [Japanese guidelines for HIV infection]. PMID- 12722300 TI - [The Japanese Respiratory Society guidelines for management of community-acquired pneumonia in adults]. PMID- 12722301 TI - [Prevention of nosocomial infection of tuberculosis]. PMID- 12722303 TI - [Guideline for STD]. PMID- 12722302 TI - [Statement on the treatment for atypical mycobacterial diseases (the Japanese Society for Tuberculosis)]. PMID- 12722305 TI - [Guide for immunization]. PMID- 12722304 TI - [Guidelines in the management of Helicobacter pylori infection in Japan]. PMID- 12722306 TI - [Appropriate use of antimicrobial agents]. PMID- 12722308 TI - [Cephem antibiotics]. PMID- 12722307 TI - [Penicillins]. PMID- 12722309 TI - [Carbapenem antibiotics]. PMID- 12722310 TI - [Macrolides]. PMID- 12722311 TI - [Quinolone]. PMID- 12722313 TI - [Anti-tuberculosis drugs]. PMID- 12722312 TI - [Other antimicrobials (linezolid, quinupristin/dalfopristin)]. PMID- 12722314 TI - [Antifungal drug]. PMID- 12722315 TI - [Antiprotozoal drugs]. PMID- 12722316 TI - [Antiretrovirals]. PMID- 12722317 TI - [Antiviral therapy for chronic viral hepatitis B and C]. PMID- 12722318 TI - [Anti-herpetic chemotherapeutic drugs]. PMID- 12722319 TI - [Antiviral agents for influenza]. PMID- 12722320 TI - [Macrolide]. PMID- 12722321 TI - [Fosfomycin]. PMID- 12722322 TI - [New effects of antimicrobial agents other than antimicrobial activity]. PMID- 12722323 TI - [Imaging diagnosis for intralobar pulmonary sequestration by subclassification of CT findings in bronchoalveolar structures]. AB - We examined the chest CT findings in 12 cases of intralobar pulmonary sequestration. We classified 4 subtypes by evaluating bronchial and alveolar structures, thus: type A (3 cases), mild cylindrical dilatation of the bronchial structure and hyperlucent alveolar structure; type B (3 cases), marked cylindrical dilatation of the bronchial structure and hyperlucent alveolar structure; type C (2 cases), multicystic dilatation of the bronchial structure and alveolar structure without hyperlucency; and type D (4 cases), multicystic dilatation of the bronchial structure and absence of any alveolar structure. All 77 cases (present and previously reported cases) with CT-documented intralobar pulmonary sequestration could be classified into 4 subtypes: type A 9%, type B 34%, type C 19%, and type D 38%. We concluded that these 4 types were useful for the radiological diagnosis of intralobar pulmonary sequestration. PMID- 12722325 TI - [The effects of pulmonary rehabilitation in elderly patients]. AB - Pulmonary rehabilitation is one of the most important treatment modalities for patients with chronic lung disease. To determine the effects of an outpatient pulmonary rehabilitation program for the older members (aged 77 +/- 3 years; seniors) of an elderly patient group with chronic lung disease, we prospectively compared the degrees of improvement of lung function, dyspnea, daily activities and exercise tolerance (6-minute walking distance) after a 9-week rehabilitation program in 27 seniors with chronic lung disease (COPD 18, post-tuberculosis lung disorders 8, lung fibrosis 1; %FEV1 50.9 +/- 17.1%) with that in disease- and %FEV1-matched younger members (aged 70 +/- 2 years; juniors; %FEV1 49.3 +/- 16.1%) of the elderly patient group. All patients performed supervised weekly outpatient exercise and education activities for 9 weeks and a home exercise regimen. Assessments were made before and after the program. Twenty-three of the seniors and 25 of the juniors completed the program. There was no significant difference in the withdrawal rates between these groups. Although lung function and blood gas data had not changed significantly after rehabilitation, the clinical symptoms and the 6-minute walking exercise improved significantly in both groups (Baseline Dyspnea Index focal score: +1.3 +/- 0.9 in the seniors and +0.6 +/- 0.9 in the juniors: 6-minute walking distance: +/- 52 m and +/- 62 m, respectively) and the improvement of the Baseline Dyspnea Index focal score was significantly greater in the seniors than in the juniors. We observed the patients after they had followed the program for 2,000 days and found that the continuation ratio of rehabilitation in the seniors was far inferior to that in the juniors (continuation ratios for 1.2 and 3 years in the seniors were 50.5, 18.0 and 0%, respectively, and in the juniors, 79.5, 66.2 and 61.5%). We concluded that, although the senior elderly patients could benefit from the pulmonary rehabilitation program, it is difficult to maintain this benefit for many years. PMID- 12722324 TI - [Prognostic significance of serum uric acid in patients with chronic obstructive pulmonary disease receiving home oxygen therapy]. AB - Home oxygen therapy (HOT) not only prolongs life expectancy but also improves quality of life. Serum uric acid (UA), the final product of purine catabolism, has been shown to be increased in the hypoxic state. To elucidate the prognostic significance of serum UA in patients with chronic obstructive pulmonary disease (COPD) receiving HOT, we assessed the ratio between the serum concentration of UA and creatinine (Cr) in 91 outpatients with COPD. During a mean follow-up period of 31 months, 24 patients died of acute exacerbation of COPD. The delta UA/Cr ratio was calculated as the percent changes in serum UA/Cr during HOT. delta UA/Cr was increased in non-survivors, but not in survivors, and was negatively correlated with the nadir of oxyhemoglobin saturation (r = -0.32, p < 0.01). Of the clinical and laboratory variables, only the delta UA/Cr ratio was found to be independently related to mortality by a multivariate Cox proportional-hazards analysis. The Kaplan-Meier survival curves divided into values below and above the median value (9.7%) of this ratio demonstrated that mortality was significantly higher among patients with high values than among those with low values (log-rank test: p < 0.05). We conclude that the delta UA/Cr ratio appears to be a reliable marker of prognosis, and may be useful for the long-term follow up of outpatients with COPD receiving HOT. PMID- 12722326 TI - [Clinical properties of community-acquired pneumonia in patients with asthma]. AB - PURPOSE: Many reports were found on the clinical properties of community-acquired pneumonia. The clinical properties of community-acquired pneumonia in patients with asthma have not been elucidated, and we therefore investigated such properties. MATERIALS AND METHODS: Asthmatic patients who required hospitalization for community-acquired pneumonia from the beginning of 1989 through the end of 2001 were enrolled in this study. We performed the study in a retrospective manner. Patients were divided into two groups based on severity of their asthma (mild to moderate asthma vs severe asthma), and we studied the clinical properties of the pneumonia. RESULT: No significant difference was seen in body temperature, white blood cell counts, or CRP value on admission between the two groups. No significant difference was seen in the resolving period of the pneumonia. The frequency of common pathogens (Streptococcus pneumoniae + Haemophilus influenzae) was lower in patients with severe asthma. Asthmatic patients not taking daily oral corticosteroids were divided into two groups based on whether or not they were using a inhaled corticosteroid, and we examined the frequency of pathogendetection. The percentage of common pathogens was almost the same in the two groups. CONCLUSION: The frequency of common pathogens was lower in patients with severe asthma than in those with mild to moderate asthma. This fact is worth considering when empiric therapy for pneumonia is performed in patients with asthma. Inhaled corticosteroid therapy seems to have no influence on the pathogens of pneumonia in patients with asthma. PMID- 12722327 TI - [Cavitary pulmonary metastasis from bladder cancer: a case report]. AB - A 69 year-old [correction of 63] man who had had a radical cystectomy for bladder cancer was admitted to our hospital because of hemosputum and right femoral pain. His chest radiograph and computed tomogram showed a mass shadow with a cavity in the left upper lung field. Sputum cytology showed class V squamous cell carcinoma and a bone scintigram showed right femoral metastasis. Despite radiotherapy to the left upper lung and the right femur, the patient's condition worsened, and he died of respiratory failure after hospitalization for about 1 month. At autopsy, pathologic studies of lung cancer revealed mixed-type transitional cell carcinoma, squamous cell carcinoma and adenocarcinoma. A diagnosis of metastatic lung cancer from bladder cancer was made. Cavitating pulmonary metastasis is uncommon. We report a rare case of pulmonary metastasis from bladder cancer, with mixed-type histopathology at both primary and metastatic sites. PMID- 12722328 TI - [Bronchiolitis obliterans organizing pneumonia after irradiation therapy for breast cancer]. AB - We report three cases of bronchiolitis obliterans organizing pneumonia (BOOP) that developed after irradiation therapy following breast cancer. All patients presented with cough and fever for 3 to 10 months after the completion of irradiation. Chest radiographs and computed tomography (CT) in all three patients demonstrated a consolidation outside the irradiated fields. Their laboratory data revealed increased C-reactive protein and increased erythrocyte sedimentation rates. Transbronchial lung biopsy was performed in all patients, and plugs of granulation tissue in the bronchioles and interstitial infiltration by mononuclear cells were found. Corticosteroid treatment resulted in rapid clinical improvement. BOOP was diagnosed from the histological and clinical findings. Although the etiology of BOOP still remains unknown, there may be a subgroup of such patients in whom the BOOP is induced by irradiation for breast cancer. These cases were assumed to be in a series of reported cases of BOOP primed by radiotherapy. PMID- 12722329 TI - [A case of Mycobacterium avium pulmonary disease detected as multiple small nodular shadows and diagnosed by CT guided transbronchial biopsy with ultrathin bronchoscopy]. AB - A 65-year-old man was admitted to our hospital because multiple small nodular shadows measuring five to 12 mm in diameter in both lung fields were noticed on chest radiographs and CT scans during a regular checkup for pulmonary emphysema. The levels of ESR and CRP were within normal limits, but those of CEA and CYFRA were slightly elevated. Mycobacterium avium (M. avium) was detected in sputum cultures and in a sputum examination using PCR. Non-necrotizing granulomas were detected in the specimens obtained from a lesion of the right S6b by CT-guided transbronchial biopsy using an ultrathin bronchoscope after navigation with virtual bronchoscopy. On the basis of these findings, M. avium pulmonary disease was diagnosed. The patient is being followed up, but because he has no symptoms, without medication. To our knowledge, there have been no previous reports of M. avium-intracellulare complex pulmonary disease, which is characterized by multiple small nodular shadows. It is speculated that multiple centrilobular lesions developed simultaneously without extension to the bronchial walls, leading in this case to multiple small nodules in both lung fields. PMID- 12722330 TI - [A case of pulmonary hamartoma and sclerosing hemangioma in the same lung]. AB - A 49-year-old woman was referred to our hospital because abnormal masses had been found in the left lung on chest radiography. This examination and CT scanning on admission both revealed a 12 x 8 mm mass in the left S5 and a 31 x 25 mm mass in the left S8. Calcification was not found in the tumors. Bronchoscopy showed narrowing of the left B5 and B8 bronchi by irregularity of the mucous membrane. No malignant cells were found in the cytological specimens, which were obtained by brushing and washing of the bronchi. Although the radiological findings suggested benign tumors, surgical resection was performed to confirm the diagnosis. The tumor in the left S5 was diagnosed as a hamartoma because it contained cartilage, lipoid tissue and lymphatic follicles. The tumor in the left S8 was diagnosed as a sclerosing hemangioma because vasculo-capillary formation by the cells with epithelial origin was seen. The pathological diagnosis of the two tumors differed, but a similar mechanism of tumorigenesis was suggested. PMID- 12722331 TI - [A case of silicoproteinosis with pneumothorax]. AB - A 46-year-old man was admitted because of an increasingly severe cough and dyspnea on exertion. For 13 years, he had inhaled sand containing 100% crystalline silica (SiO2). Chest radiographs revealed right pneumothorax and diffuse small nodular and ground-glass opacities in both lungs (especially in the upper lung fields). A chest CT scan disclosed several bullae in both upper lobes, and an open lung biopsy was performed along with resection of these bullae. Subsequently, silicotic nodules containing silica and PAS-positive materials were recognized in the alveolar spaces in the histological findings, and a diagnosis of silicoproteinosis was made. We have reported on this case of silicoproteinosis with pneumothorax which progressed for over one year and which showed unusual radiological findings dissimilar to those of primary pulmonary alveolar proteinosis. PMID- 12722332 TI - [Acute lung injury caused by inhalation of waterproofing spray]. AB - A 55-year-old man was admitted to our hospital because of severe dyspnea 30 minutes after inhalation of waterproofing spray. He had used the spray outdoors and had then smoked a cigarette with spray-contaminated fingers. Chest radiography and computed tomography (CT) revealed diffuse ground glass opacities in both lungs. In pulmonary function tests, the lungs showed a moderately decreased diffusing capacity and there was slight hypoxemia. Transbronchial lung biopsy specimens demonstrated extensive alveolitis and marked eosinophil migration. Without any specific treatment, the patient recovered clinically in 4 days. We speculated that acute lung injury in this patient may have been induced by not only direct inhalation of the waterproofing spray itself, but also by inhalation of spray by-products resulting from decomposition due to heat. When waterproofing spray is used, precautions should be taken to avoid both inhalation and heating of the fumes. PMID- 12722333 TI - [A case of pulmonary low-grade B cell lymphoma (MALT type) presenting seven years after gastric lymphoma resection]. AB - A 71-year-old man was admitted to our hospital because of systemic edema and exertional dyspnea. Chest radiographs revealed infiltrative shadows in both lung fields, pleural effusion, and pericardial effusion. Seven years before, he had undergone gastric surgery for a gastric ulcer with lymphoid hyperplasia. In the pathologic diagnosis based on the percutaneous lung biopsy, hyalinizing granuloma was suspected. For a more thorough diagnosis, the patient was subjected to an open lung biopsy, and the final diagnosis was low-grade B-cell lymphoma of the MALT (mucosa-associated lymphoid tissue) type. Gallium scintigraphy showed accentuated accumulation in the left neck and hypothyroidism was present. Histologic re-examination of the resected stomach revealed infiltration of centrocyte-like cells and lymphoepithelial lesions, compatible with the pathologic features of MALT lymphoma. We considered that the gastric neoplasm and the pulmonary, pleural, and thyroid tumors of MALT lymphoma had occurred seven years apart in this case. Thyroid hormone replacement and CHOP therapy improved the symptoms and decreased the lung tumor size by 73%. MALT lymphomas tend to remain localized for a long period. The multiorgan involvement seen in this case is rather rare. PMID- 12722334 TI - [Summer-type hypersensitivity pneumonitis in monozygotic twins]. AB - We encountered a pair of monozygotic twins with summer-type hypersensitivity pneumonitis. Patients 1 and 2 were 24-year-old men who worked in the same place and shared the same room, which was built of wood. In August, patient 1, a non smoker, was referred to our hospital because of coughing, fever and dyspnea. In November, the same symptoms appeared in patient 2, who was a smoker. The difference between the onset times in patients 1 and 2 was suspected to be due to cigarette smoking, because their occupations and other circumstances were the same and they were monozygotic twins. PMID- 12722335 TI - [Electromyographic activity of human triangularis sterni muscles during thoracic rotation]. AB - The triangularis sterni muscles (TS) have been recognized as expiratory muscles, but there is no information on their postural activity or their phasic electromyographic (EMG) activity during thoracic rotation. We examined the EMG activity of the right TS using fine-wire electrodes, in 5 healthy male subjects. With subjects breathing through a mouthpiece, airflow, raw and moving-averaged EMG signals were sampled with a computer during thoracic rotation and resting, accompanied by breathing with or without voluntary thoracic rotation. Maximum EMG (EMGmax) was defined as the greatest moving-averaged EMG activity recorded from each subject during the slow expiratory maneuver from functional residual capacity to residual volume. With leftward thoracic rotation without breathing, the tonic EMG activity of the right TS was observed in all subjects. During resting breathing without thoracic rotation, there were respiration-related phasic activities on TS EMG during expiration in all subjects. With leftward thoracic rotation, the respiration-related phasic activities on TS EMG significantly increased from 8.2 +/- 3.5 (mean +/- SE) %EMGmax to 15.2 +/- 6.1 %EMGmax. On the other hand, with rightward thoracic rotation, it decreased to 3.0 +/- 1.0 %EMGmax. We concluded that 1) TS has a postural function like that of the intercostal muscles, and that 2) the respiratory function of TS is affected by thoracic rotation. PMID- 12722336 TI - [Case report of pulmonary epithelioid hemangioendothelioma]. AB - A 45-year-old man was admitted to our hospital because of multiple nodular shadows in the right upper field of a chest radiogram taken at a regular medical checkup. He underwent open lung biopsy. The lung tumor found was diagnosed histologically as pulmonary epithelioid hemangioendothelioma. The tumor cells showed positive staining for CD34 and factor VIII-related antigen. Pulmonary epithelioid hemangioendothelioma (PEH) is a rare lung tumor, of which only 40 cases, including the present case, were reported between 1983 and 2002 in Japan. PEH is a progressive, low-grade malignant tumor that originates from hemangioendothelial cells. In chest radiography or CT scanning, PEH is usually discovered incidentally as multiple nodular shadows. Many cases of PEH are diagnosed by open lung biopsy or thoracoscopic biopsy. No standard therapy for PEH has yet been established, other than resection of a solitary lesion. The present patient has been followed without treatment for five-and-a-half years, and is still alive with no symptoms. PMID- 12722337 TI - [Pathophysiology and treatment of a plastic anemia: useful markers for the diagnosis of immune pathophysiology]. PMID- 12722338 TI - [Trends in the therapy for low-grade non-Hodgkin lymphoma]. PMID- 12722339 TI - [Co-expression of major and minor bcr/abl in chronic myelogenous leukemia]. PMID- 12722340 TI - [Gene expression profiling of de novo CD5-positive diffuse large B-cell lymphoma]. PMID- 12722341 TI - [Congenital plasmin inhibitor deficiencies: basic concepts and clinical applications]. PMID- 12722342 TI - [vWF-cleaving protease/ADAMTS 13]. PMID- 12722345 TI - [Recent medical treatment for advanced hepatocellular carcinoma]. PMID- 12722344 TI - [Disappearance of CD 20 after treatment with rituximab of mantle cell lymphoma]. AB - A 60-year-old female visited our hospital in May 2001 because of systemic lymphadenopathy. Her white blood cell count was 25,510/microliters with 93% of lymphocytes. Bone marrow aspiration revealed that 86% of nucleated cells were lymphocytes. Lymphocytes in the peripheral blood and bone marrow were positive for CD 5, 19, 20, and sIgx and negative for CD 23. FISH analysis detected the chimeric bcl 1/IgH fusion gene. Immunohistochemistry of a biopsied lymph node revealed that lymphoma cells were positive for cyclin D 1. Mantle cell lymphoma (MCL) was diagnosed at clinical stage IV A. Although a partial remission was obtained after CHOP plus rituximab therapy, the patient's disease recurred in March 2002 and she died in spite of salvage therapy including rituximab. Immunohistochemistry of the bone marrow cells after salvage rituximab therapy revealed that lymphoma cells were still positive for CD 5 and cyclin D 1, but negative for CD 20 and sIgx. We could not exactly determine how frequently CD 20 expression becomes negative in B-cell lymphomas after treatment with rituximab. We found only two reported cases that suggested rituximab down-regulated CD 20 expression in MCL. We herein describe a case of MCL with very notable clinical features. PMID- 12722343 TI - [Therapy-related acute myelogenous leukemia (AML-M6) with add(11) (q23) and del(20) (q11.2) developing via myelodysplastic syndrome after chemotherapy for malignant lymphoma]. AB - A 62-year-old woman was diagnosed as having malignant lymphoma, diffuse large B cell type. She underwent chemotherapy with the standard dose of CHOP and MINE regimens, resulting in complete remission. Four months later, the myelodysplastic syndrome of RA (refractory anemia) with pancytopenia developed and rapidly progressed to acute myelogenous leukemia (AML-M6) in 4 months. Cytogenetic analysis for the bone marrow specimens of both periods of MDS and AML-M6 revealed complex karyotypic abnormalities involving chromosome 5, 7, 11q23 and 20q11.2. Neither rearrangement of the MLL gene by Southern blot analysis nor tandem duplication of MLL gene by RT-PCR technique was detected. The patient was died from progression of leukemia and pneumonia. The autopsy showed no residual disease of lymphoma-related disease. PMID- 12722346 TI - [Progress in surgical therapy for advanced hepatocellular carcinoma]. PMID- 12722347 TI - [A case of gastric fundic polyps during long-term treatment of reflux esophagitis with omeprazole]. PMID- 12722348 TI - [A case of steroid dependent ulcerative colitis (total colitis type) treated by combined use of germinated barley foodstuff]. PMID- 12722349 TI - [A serious case of pseudomembranous colitis accompanied by toxic megacolon in patient of paralytic ileus]. PMID- 12722350 TI - [A case of Cronkhite-Canada syndrome showing specific nail deformity after remission with prednisolone]. PMID- 12722351 TI - [Leiomyosarcoma with osteoclast-like giant cells in the small intestine]. PMID- 12722352 TI - [A case of advanced hepatocelluar carcinoma achieving a complete response following the hepatic arterial infusion chemotherapy and subsequent lipiodolization]. PMID- 12722353 TI - [An acute hepatitis case domestically infected with a hepatitis E virus whose nucleotide sequence showed a high similarity to that from a domestic swine in Japan]. PMID- 12722354 TI - [Prednisolon-induced acute pancreatitis in an adult patient with idiopatic thrombocytopenic purpura]. PMID- 12722355 TI - [A case of ileal ulcer in the blind pouch syndrome diagnosed by retrograde ileoscopic examination]. PMID- 12722356 TI - [Development of the concept and governmental program of a rehabilitation system for participants in military operations]. PMID- 12722357 TI - [Training of surgeons in the emergency medical care university department]. PMID- 12722358 TI - [Loss of visual function as the result of gunshot injuries of the eye]. AB - The article presents the clinical-and-statistical analysis of causes of visual function loss occurred after the gunshot injuries of eyes. The main cause of sight loss is the eye destruction as the result of gunshot injury (61.1%). Because of the unsuccessful treatment of posttraumatic changes in eyeball the injured eyes were removed in 38.9% of the casualties. In most cases the fragmentation and mine-explosive trauma were the cause of eye destruction. In the cases of eyeball destruction enucleation and evisceration were the main surgical interventions. In the therapy of posttraumatic changes in the injured eye (flaccid inflammatory process without tendency towards its resolution, painful syndrome as well as eyeball atrophy and subatrophy) enucleation was performed. Improvement of conservative methods in therapy of posttraumatic complications could contribute to decrease in the number of casualties with sight loss after the eye gunshot injures. PMID- 12722359 TI - [Method of excimer laser personalized ablation in astigmatism correction for the patient with sequelae of corneal penetrating wounds]. AB - Aberrations of 25 eyes of patients with posttraumatic corneal scars and artiphakia were investigated using OPD-scan aberrometer. All patients underwent LASIK using the segmental and personalized ablation technology. Clinical and functional results were followed-up during up to 18 months after the surgery. Personalized ablation technology can be safely and effectively used in correction of sphere, cylinder and high order aberrations and allows to achieve maximally high refractive and functional results in the patients with refraction anomaly after eye trauma. Today in most cases such effective results can't be achieved by any other correction method. PMID- 12722360 TI - [Principles of treatment and prophylaxis of chronic prostatitis]. PMID- 12722361 TI - [Treatment of visceral pathology in patients with trauma]. AB - The problem of visceral pathology in patients with trauma that was intensively investigated by the scientists during the period of Great Patriotic War and in post-war period remains actual in our days. The purpose of investigation is to study the possibilities, peculiarities and efficiency of diagnosis, treatment, rehabilitation and medical examination of the casualties suffering from visceral pathology in different periods after trauma in conditions of therapeutic hospital as well as the peculiarities of their regular medical checkup at the post hospital stage. It will allow to use more widely the therapeutic beds for the complex medical support of this patient group. 692 patients admitted in different periods after mild mechanical trauma complicated by different visceral diseases were investigated in conditions of therapeutic hospital. After the complex investigation the following conclusion was drawn: the casualties with mild closed injury complicated by visceral pathology even in the early posttraumatic period can be send to the therapeutic hospital where the adequate medical care will be rendered. It is especially important during the simultaneous mass admission of the casualties in the time of natural disaster and catastrophes, in conditions of military operations when the surgical subunits of medical evacuation stages will be full of persons who require the active resuscitation and surgical intervention. Complex medical support of the casualties with visceral changes in the late periods after trauma is the many-sided process connected with solution of therapeutic, rehabilitation, expert-and-diagnostic problems that are fully in the sphere of specialists of therapeutic institutions. PMID- 12722362 TI - [Significance of microelements in the development of typical pathological process in the presence of ischemic heart disease]. PMID- 12722363 TI - [Prophylaxis of pneumonia occurring outside the hospital in organized groups]. PMID- 12722364 TI - [The problem of otomycosis treatment]. PMID- 12722365 TI - [Combined infection of abdominal typhoid and hepatitis A and E]. PMID- 12722366 TI - [Hypobaric variant of interval hypoxic training in aerospace medicine]. PMID- 12722367 TI - [Graduates of the year of "Stalingrad"]. PMID- 12722368 TI - [Medical service provided prisoners of war during the period of the Stalingrad offensive operations]. PMID- 12722370 TI - [Correlation between the expression of mRNA of histones H2B and H4 and mRNA of kinin receptors B1 and B2 assessed by QRT-PCR in vulvar cancer]. AB - OBJECTIVES: Histones are involved in process of proliferation and differentiation of the cells. In many lesions the expression of mRNA of histones is a marker of proliferation. The expression of kinin receptors B1 and B2 coexist with inflammatory processes, but in some studies the expression of these receptors was found to influence the growth and differentiation of different cells. DESIGN: In search of proliferation markers in squamous cell vulvar cancer we have analysed the correlation between the expression of the mRNA of histone H2B and H4 and mRNA of kinin receptors B1 and B2. MATERIALS AND METHODS: The specimen obtained from 46 women operated for squamous cell vulvar cancer stages I to IV according to FIGO were analysed. RNA was extracted and the number of mRNA copies were assessed by QRT-PCR using ABI PRISM 7700 (TaqMan). RESULTS: The results obtained in this study indicate a statistically significant correlation between mRNA B1 and mRNA H4 (p < 0.01), B2 and histone H4 (p < 0.01) B1 and histone H2B (p < 0.01). CONCLUSIONS: The expression of mRNA of histones and kinins receptors may reflects the dynamics of neoplastic growth in vulvar cancer. PMID- 12722371 TI - [Abrikosov's tumor of the vulva--a case report]. AB - A rare case of Abrikosov's tumor of the vulva has been reported. A 49 year old postmenopausal woman was admitted to hospital with a slow growing tumor of the left greater pudendal lip. The patient underwent tumorectomy. Histologically GCM- granular cell myoblastoma known as the Abrikosov's tumor was diagnosed. As the tumor was of no potential malignancy the patient did not require any additional treatment. We present our experience and the review of current literature. PMID- 12722372 TI - [Surgical treatment of vulvar cancer in 101-year old patient--a case report]. AB - OBJECTIVES: Vulva cancer makes up 2.5-5% of all malignant neoplasms developing within the female genital organs. The average age of patients with vulvar cancer is 60 years, and most cases occur between 70-80. The great majority of cases (95%) belong to squamous cell carcinoma with endophytic or exophytic growth, for example carcinoma verrucosum. It is malignant neoplasms showing a significant histologic maturity, which however, does not cause metastases. STUDY DESIGN: This is a case report concerning a 101 year old patient with vulvar cancer treated surgically. CONCLUSIONS: In cases of vulvar cancer, surgical treatment is still the method of choice. The surgical removal of the tumor together with the surrounding tissues proved to be sufficient. PMID- 12722373 TI - [Clinical analysis of 105 cases of vulvar cancer]. AB - OBJECTIVES: Vulvar cancer has a low frequency (4-5%) compared to other genital cancers. It should be considered as a skin tumor and its detection is possible in an early stage. However, because of patients' and doctors' delay, one in three vulvar cancers is not treated before an advanced stage. DESIGN: To estimate clinical and histopathological features of vulvar cancer. MATERIAL AND METHOD: We analysed 105 women with a primary malignant vulvar disease treated operatively in our Dept. in the years 1991-2002. In the group there were 82 invasive tumours (80 squamous and 2 solid) and 23 intraepithelial cancers. RESULTS: The mean age in the group of intraepithelial cancer was 53.2 +/- 13.2 years, and in the group of invasive disease 65.0 +/- 11.7 years. The most common symptoms were ache (40.0%) and pruritus of the vulva (35.2%). 33.3% women were asymptomatic. Most of cancers were localised on the large pudendal lip (71.4%). Inguinal node dissections performed on invasive cancer patients showed negative nodes in 57 cases and positive in 25 cases. Sensitivity, specificity, positive and negative predictive value for clinical detection of metastatic cancer in the inguinal nodes were 60%, 82.4%, 60%, 82.4% respectively. The distribution by stage included Stage 0- 21.9%; Stage I--19.0%; Stage II--31.4%; Stage III--11.5%; and Stage IV--19.0%. The commonest surgical procedure was a simple vulvectomy with a bilateral inguinal lymphadenectomy. CONCLUSIONS: Vulvar squamous cell carcinoma predominantly affects older women. It benefits from early detection, and their morbidity is decreased by early treatment. It is necessary to educate patients to improve the detection of vulvar cancer. PMID- 12722374 TI - [Function of the lower urinary tract in patients after treatment of cervical cancer]. AB - OBJECTIVES: Irritation of lower urinary tract--bladder and urethra--is seen as a common risk factor for radiotherapy treatment. Is it possible to reduce miction disorders and urinary incontinence? DESIGN: Functional assessment of the lower urinary tract in women treated with surgery and radiotherapy. MATERIALS AND METHODS: Authors evaluated 25 patients diagnosed with uterus neck cancer treated with surgery or radiotherapy during 1998-2001. Urodynamic assessment was based on uroflowmetry, residual urine volume measurement and cystometry results. RESULTS: Lower urinary tract ailments were present in all 25 patients, in 2 of them they were only temporary--during recovery after surgery. During miction symptoms were reported by 23 patients, 20 patients reported symptoms between mictions. During and after treatment all patients reported the increase in symptoms and changes of urodynamics parameters. CONCLUSIONS: Function of the lower urinary tract are disturbed whilst radiotherapy treatment of uterus neck cancer. Functional diagnostics is important in oncological treatment design and should be evaluated separately for each patients. PMID- 12722375 TI - [Lymph node metastasis as a prognostic factor in cervical carcinoma]. AB - MATERIAL AND METHODS: 499 patients with cervical carcinoma at stage I and IIa after radical hysterectomy were included in the study. Diagnosis was based on gynecological examinations and cervical biopsies. Clinical staging was determined by FIGO classification. Pelvic lymph nodes were routinely removed on hysterectomy. RESULTS: Metastatic nodes were observed in 26.3% (131 patients). We found no metastatic nodes at stage Ia. In the group of 410 patients with stage Ib cervical cancer metastases in lymph nodes were found in 24.6% (101 patients). In the group of 78 patients with stage IIa cervical cancer metastatic nodes were observed in 38.5% (30 patients). In our finding metastases were located in one group of lymph nodes in 64.4% (64 patients) with stage Ib and 43.3% (13 patients) with stage IIa. Metastatic involvement of more than one group of lymph nodes was observed in 36.6% (37 patients) of stage Ib and 56.7% (17 patients) of stage IIa. The most frequent pattern of lymph nodes metastatic involvement comprised common iliac and obturatorious nodes. 5 year survival in the group without metastases in lymph nodes was estimated at 82.2%, and in the group with nodal metastases--50.8% (p = 0.005). CONCLUSIONS: 1. Metastases to pelvic lymph nodes are significant prognostic factor of long-term survival in patients with cervical cancer. 2. Patients with metastases in lymph nodes and no subsequent postoperative radiotherapy had significantly worse long-term survival. PMID- 12722376 TI - [The effectiveness of photodynamic diagnosis method in diagnosis of cervical lesions]. AB - OBJECTIVES: The aim of this study was to evaluate Photodynamic Diagnosis (PDD) effectiveness among patients with cervical lesions and comparison to effectiveness of colposcopy. MATERIAL AND METHODS: 68 patients with cervical lesions underwent cytologic examination, colposcopy and PDD which were followed by histologic examination. RESULTS: In 41 patients with Pap smear gr. II or IV the result of Pap smear, colposcopy, PDD and histologic result were correlated. Among 27 patients with Pap smear gr. III the conformity of the final pathologic result with colposcopic examination was 55.6% and positive fluorescent effect during PDD was 63% in patients with Pap smear gr. III. CONCLUSION: The efficacy, specificity and sensitivity of PDD are similar to those of colposcopy in diagnosis of Pap smear gr. III patients. PMID- 12722377 TI - [The results of screening program for cervical cancer in Lodz]. AB - OBJECTIVES: Despite effective screening methods, most often tumor of the urogenital tract is still cancer of the uterine cervix. Proven in many countries the value of widespread screening of the precancerous lesions and cervical cancer justifies carrying out such prophylactic tests in Poland. DESIGN: Assessment of results of screening as a method of early detection of cancer of the uterine cervix among inhabitants of Lodz area. MATERIAL AND METHODS: Study was done between 1.07.2000 and 31.12.2000 among 5,000 women aged from 30 to 59 years, inhabitants of Lodz area. Smears were evaluated in five-grade Papanicolaou scale and according to TBS system. In cases of detection of erosion of the uterine cervix or abnormal results of cytological test (> or = III degree) biopsies were taken to make pathological diagnosis. RESULTS: 459 smears were qualified to group I according to Papanicolaou (9.18%), 4435 smears to group II (88.7%), 38 smears to II/III (0.76%), 65 smears to group III (1.3%), 2 smears to IV (0.04%) and 1 smear to V (0.02%). Due to histopathological verification of the cytological results 53 low grade cervical dysplasia (1.06%) were diagnosed, 8 dysplasia of medium grade (0.16%), 7 high grade dysplasia (0.14%), 3 pre-invasive cancers (0.06%) and 3 invasive cancers (0.06%). CONCLUSIONS: Population of inhabitants of Lodz area aged 30 to 59 years old is characterised by a high frequency of incidence of pre-cancerous conditions and invasive cervical cancer. Number of detected pathologies of the uterine cervix proves purposefulness of this project. PMID- 12722379 TI - [Analysis of epidemiologic risk factors for endometrial cancer]. AB - The aim of this study was an analyse of several risk factors for women with endometrial cancer and their influence on overall survival. MATERIALS AND METHODS: A retrospective analysis covered 160 women treated with the primary surgery and adjuvant radiotherapy because of endometrial cancer. Age, lifestyle, age of menarche and menopause, parity, body size, hypertension and diabetes mellitus were determined. RESULTS: The 5-year survival rate were: 87% for the women younger then 60 and 70% for the older ones (p = 0.0026), 81% for the city women and 73% for the country women (p = 0.2034), 91% for the educated women and 70% for the women with elementary education (p = 0.0102), 85% for 1-2 x parous women and 65% for nulliparous (p = 0.0433), 79% for women without miscarriage and 63% for women with 2 or more miscarriages (p = 0.3265), 82% for women with menarche above age of 15 and 66% with menarche under age of 13 (p = 0.1989), 82% for women without obesity, diabetes, hypertension and 72% for women with hypertension, 64% with obesity, 50% for women with all analysed diseases (p = 0.0104). CONCLUSION: An analyse show that age, lifestyle, parity, body size and hypertension modify the overall survival of women with endometrial cancer. PMID- 12722378 TI - [Frequency of HPV infection of the uterine cervix among perimenopausal women in Wielkopolska Region]. AB - OBJECTIVES: Human papillomavirus (HPV) is one of the main causes of cervical neoplasia. There is now consistent evidence that Human Papillomavirus (HPV) has a causal role in the etiology of cervical cancer and that sexual habits and reproductive/hormonal factors are associated with the risk of invasive cervical cancer. In our investigation we estimated the frequency of HPV infection in uterine cervix in women gathered in perimenopausal and postmenopausal groups. MATERIALS AND METHODS: We screened 90 women in western Poland to obtain cytological smears, and tested all smears for 33 types of HPV with a polymerase chain reaction-based system. All women were classified according to age into three groups. RESULTS: HPV infections peaked in the group of 45 to 49 year old (48.5% using universal starters) and decreased in 56 year or olders (26.1% using universal starters) with predominantly non-cancer-associated types of HPV and uncharacterized HPV types. CONCLUSIONS: We confirm the decline of HPV infection with age. But note increased prevalence after menopause, which could be related to a second peak of HSILs, an observation that warrants further investigation. At least 80% of HPVs involved in cervical carcinogenesis in this population have been characterized. Polyvalent vaccines including the main cancer-associated HPV types may be able to prevent most cases of cervical disease in this region. PMID- 12722380 TI - [Microvessel density index as a prognostic factor in a low histological differentiation stage of endometrial carcinoma]. AB - OBJECTIVE: Angiogenesis in malignant tumors is a prognostic factor associated with tumor growth and metastasis. Studies of angiogenesis in breast, prostate and lung cancer showed that neovascularisation correlates with likelihood of metastasis and recurrence. Our study was to evaluate microvessel density as a prognostic factor in endometrial cancer. METHODS: Between 1995-1999, 58 women were treated for endometrial carcinoma. The primary treatment consisted of total abdominal hysterectomy, bilateral salpingoophorectomy and pelvic lymphadenectomy. The microscopic examination of paraffin blocks showed the areas of the deepest myometrial invasion. The microvessels within the invasive cancer were highlighted by means of immuno-cytochemical staining to detect CD-31 antigen. RESULTS: Microvessel count was related to likelihood of recurrence. We found statistically significant differences between patients who died after operation and patients with nonrecurrence process. All patients were in the same stage and grade of endometrial carcinoma. CONCLUSION: Microvessel density index seems to be an important factor for planing postoperation treatment in endometrial carcinoma. PMID- 12722381 TI - [Evaluation of selected risk factors in patients operated for uterine body carcinoma]. AB - OBJECTIVES: Uterus body carcinoma ranks among to the most common malignant neoplasms. It is of special interest what kind of factor influences on the survival span of treated patients. DESIGN: Evaluation of selected risk factors such as: age, obesity, clinical stage, type and histologic differentiation and proliferation depths on the 5-year survival span. MATERIALS AND METHODS: /Due to our research we analyzed 280 endometrium samples of patients with the diagnosis of endometrial cancer, who*.../ Analysis was made on the group of 280 endometrium carcinoma patients, who were operated in the Department of Obstetrics and Women Diseases, University School of Medicine in Bydgoszcz in 1982-2000 period, 221 of the total number of patients (78.8%) were in the cancer stage I, while 33 (11.9%) in stage II, followed by 26 (9.3%) in stage III. RESULTS: The highest percentage of 5-year survival time was observed in the group of patients under age of 50. In patients with the clinical stage I and proliferation depths under 1/2 of myometrum the percentage of 5-year survival time was 91%. Adenocarcinoma was the most common type of cancer and also with the best prognosis (81% of 5-year survival). Neither histologic differentiation G1 nor G2 have an influence on the survival time of operated patients. CONCLUSIONS: 1. The proliferation depths, histologic type of the endometrial carcinoma and clinical stage are the major prognostic factors for the uterus body carcinoma. 2. Endometrial carcinoma in the group of patients under age of 50 is connected with the higher percentage of five year survival time. PMID- 12722382 TI - [Estimation of DNA methylation level in nonendometrial uterus malignancies]. AB - OBJECTIVES: Rebuilding of genome structure leads to many pathological states including neoplastic malignancies. Rebuilding often occurs as a process caused by disturbances in gene silencing mechanism. DNA methylation pattern is one of the most important mechanisms connected to gene's silencing. Estimation of DNA methylation level in nonendometrial uterine neoplastic tissues compared to normal endometrial samples was the aim of this study. DESIGN: It was to be shown, that changes in methylation rate in promotory regions could lead to carcinogenesis in particular cell. Authors describe an analysis of DNA methylation level in nonendometrial neoplastic uterine tissues compared to DNA methylation level in normal endometrium. MATERIALS AND METHODS: Tissue samples from 9 women with tumor mixtus mesodermalis were collected. 12 samples were normal endometrium-control group. DNA was isolated from tissues and than we performed an estimation of DNA methylation levels. Than we performed a statistical analysis of results. RESULTS: The median DNA methylation level was significantly higher in neoplastic tissues than in normal endometrium. CONCLUSIONS: Authors conclude, that DNA methylation level is higher in neoplastic tissues, but does not correlate with clinical stage of the disease. PMID- 12722383 TI - [Estimation of DNA methylation level in endometrial cancer tissues]. AB - OBJECTIVES: A level of DNA methylation plays an important role in regulation of cellular gene's expression. Estimation of DNA methylation level in endometrial neoplastic tissues compared to normal endometrial samples was the aim of this study. DESIGN: It was to be shown, that changes in methylation rate in promotory regions could lead to carcinogenesis in particular cell. Authors describe an analysis of DNA methylation level in endometrial cancer tissues compared to DNA methylation level in normal or hyperplastic endometrium. MATERIAL AND METHODS: Endometrial samples from 88 women were collected. 56 of them were classified as adenocarcinoma, 20 as hyperplastic changes, 12 as normal endometrium-control group. DNA was isolated from tissues and than prepared to pm5dC and pdC. Than we performed a statistical analysis of results. RESULTS: The median DNA methylation level was significantly higher in neoplastic tissues than in normal endometrium. There was no difference between DNA methylation level between normal endometrium and hyperplastic changes. CONCLUSIONS: Authors conclude that neoplastic endometrial tissues show high DNA methylation rate compared to normal or hyperplastic endometrium. PMID- 12722384 TI - [Three dimensional sonography in the endometrial volume measurement in women with perimenopausal irregular uterine bleeding]. AB - OBJECTIVES: Evaluation of the role of three-dimensional (3D) ultrasonographic measurement of the endometrium volume in perimenopausal women with irregular uterine bleeding. MATERIAL AND METHODS: We have assessed 45 women (23 postmenopausal) with 3D transvaginal probe and automatic volume measurement (VOCAL) software using Voluson V730 scanner (Kretztechnik, Austria). Additionally, doppler blood flow indices PI, RI and PSV in uterine artery as well as endometrial thickness were measured. All ultrasound data were verified by histology of the endometrial specimens removed during D&C or hysterectomy. RESULTS: The age of the study group ranged between 43-70 years (median 53 years). The mean volume of endometrium in women with endometrial cancer was 19.9 +/- 7.5 ml. The mean volumes measured in women with endometrial hyperplasia and normal endometrium were 12.2 +/- 7.9 ml and 7.4 +/- 4.8 ml, respectively. There was a significant difference in endometrial volume between each pair of study groups (P < 0.05). Moreover, significant differences were found in mean endometrial thickness between endometrial cancer and benign but not in hyperplastic endometrium. In contrast, there were no significant differences in study groups in mean values of Doppler blood flow indices measured in uterine artery. CONCLUSION: We conclude that 3D sonography and automatic volume measurement might be used better to characterize endometrial changes in women with perimenopausal bleeding. PMID- 12722385 TI - [Clinical value of pelvic lymphadenectomy in surgical treatment of endometrial cancer]. AB - OBJECTIVES: The aim of the study was the clinical analysis of value of pelvic lymphadenectomy in surgical treatment of endometrial carcinoma. MATERIAL AND METHODS: The group of 420 women treated for of endometrial cancer in stage IA-IV by FIGO 1988 was analysed. All these women were operated in 2nd Dept. of Obstetrics and Gynecology Medical University of Gdansk between 1981-2000. The subgroup of patients with pelvic lymphadenectomy was selected. The data were obtained from case histories. RESULTS: The subgroup of patients on whom pelvic lymphadenectomy was performed was up 32.6% of all treated women (137 women). Neoplastic metastases were found in 27 cases (19.7%). All those patients were in clinical stage II-IV by FIGO 1988, and no earlier stage of endometrial cancer was found in that group. In every case myometrial invasion was more then 1/2 of myometrium. In 59.2% of neoplastic metastases high or medium differential adenocarcinoma were found. CONCLUSION: Pelvic lymphadenectomy has to be part of surgical treatment of endometrial cancer. Lymph nodes metastases are important prognostic factors. Myometrial invasion plays an important role as a risk factor of lymph nodes metastases. PMID- 12722386 TI - [Clinical assessment of sonohysterography in the diagnosis and management of abnormal uterine bleeding]. AB - OVERVIEW: Abnormal uterine bleeding is a common gynecological symptom. The curettage can asses only 50-60% of endometrium surface, thereby leaving some changes unrevealed. Sonohysterography (SHG), which combines the distention of the uterine cavity trough saline solution and ultrasound examination, allows a more exact evaluation of uterine cavity abnormalities. OBJECTIVES: To assess the value of sonohysterography in the diagnosis and management of abnormal uterine bleeding. MATERIALS AND METHODS: 67 women with abnormal uterine bleeding and uterine cavity abnormalities confirmed by transvaginal ultrasound were examined with sonohysterography. RESULTS: Sonohysterography revealed 21 cases of submucous myomas, 22 endometrial polyps and 24 cases of endometrial hyperplasia. All patients underwent diagnostic hysteroscopy to verify the results. The sensitivity and specificity for sonohysterography were both 93%. The positive and negative predictive values were 96% and 93%, respectively. CONCLUSIONS: Sonohysterography is a sensitive and specific method for diagnosis of abnormal uterine bleeding. It allows faster and less invasive diagnosis. Given the low costs of examination sonhysterography can be used as an ambulatory procedure reducing delay of treatment. PMID- 12722387 TI - [Transvaginal ultrasonography in the diagnosis of endometrial and uterine cavity changes in perimenopausal women]. AB - DESIGN: The purpose of the study was to estimate the diagnostic value of ultrasonic endometrium thickness measurement and estimation of ultrasonic endometrium qualitative features in detecting pathological changes in women in perimenopausal period. MATERIALS AND METHODS: The group of the patients consist of 242 patients in age 45-86 years, with abnormal uterine bleeding or incorrect ultrasonographic image of endometrium. In all cases transvaginal ultrasonography (TVS) and hysteroscopy were performed. RESULTS: The average thickness of endometrium in carcinoma (CA), hyperplasia (H) and polyps (P) group (properly 8.96 and 6.09 and 5.02 mm) showed essential differences in comparison with a group without changes in endometrium (3.38 mm). In group CA and H the greatest cumulation of abnormal features of ultrasonic image was ascertained. CONCLUSIONS: Ultrasonic measurement of endometrial thickness is a sensitive index in detecting cancer and pathological endometrial hyperplasia. The combination measurement of endometrial thickness and estimation of qualitative features of endometrial and uterine cavity TVS image improves results of detecting all of types of intrauterine pathology. PMID- 12722388 TI - [Assessment of the reliability of different methods in diagnosing endometrial cancer]. AB - OBJECTIVES: Endometrial cancer is the most common gynecologic malignancy. The purpose of the study was to compare the reliability of different methods for diagnosing endometrial cancer. DESIGN: A prospective study from January 1996 to September 1998. MATERIALS AND METHODS: 264 women at risk of endometrial cancer with clinical indications for hysteroscopy with curettage participated in the study. They underwent different diagnostic methods: endometrial cytology (121 cases), endometrial biopsy (150) or ultrasonography (200). The accuracy of these methods was related to the following results obtained from hysteroscopy with curettage. RESULTS: The results of the study suggest higher reliability of endometrial biopsy from cytology. The hysteroscopic diagnosis conforms with the histologic examination showed by the curettage. Endometrial cancer as the cause of post-menopausal bleeding can be excluded in cases with endometrial thickness of < or = 5 mm measured by vaginal ultrasonography. CONCLUSIONS: We concluded that the combination of hysteroscopy and endometrial biopsy may substitute for curettage and become the method of choice for the endometrium. PMID- 12722390 TI - [Risk factors for gestational trophoblastic tumors following complete hydatidiform mole]. AB - OBJECTIVES: The aim of our study was to determine the factors associated with higher incidence of trophoblastic tumours following complete hydatidiform mole. MATERIAL AND METHODS: Epidemiological and clinical factors were studied in eighty five patients with complete hydatidiform mole evacuated from 1973 to 1997 in Department of Obstetrics and Gynecology Medical University of Gdansk. Univariate and multivariate analysis were used to study of group. RESULTS AND CONCLUSIONS: In the analysis of 85 patients we found three prognostically independent factors that were associated with higher incidence of trophoblastic tumours after the complete hydatidiform mole: pre-evacuation hCG level, presence of prominent theca lutein cysts (greater than 6 cm in diameter) and molar pregnancy in the patient's past history. Persistent vomiting was a symptom of lower significance as a risk factor of trophoblastic tumour. PMID- 12722391 TI - [Externalized anus in patients with advanced ovarian cancer]. AB - OBJECTIVE: Intestinal operations in advanced ovarian carcinoma due to intestinal obstruction are still controversial. DESIGN: The aim of study was to analyse the efficacy of intestinal operation and making anus praeter in the patient with advanced ovarian carcinoma. MATERIAL AND METHODS: A total of 497 patient with ovarian carcinoma were operated in II Department of Obstetrics and Gynecology in 1990-2001. 66 patients with advanced ovarian cancer were analysed with a view to interventions for intestinal obstruction and making the anus praeter. RESULTS: In the group of 497 patients with ovarian carcinoma there were a operations of making anus praeter in 66 cases (13.28%). It was performed during first operation in 40.9%, during SLO in 22.72%, TLO in 16.67% and following operations in 19.70%. In 5 cases (7.58%) there was a liquidatio of anus praeter after an average 8.8 months after intestinal operation. CONCLUSION: Surgical intervention can be of value in removing life-threatening intestinal obstruction and to achieving longer survival with a possibility of receiving a further antineoplastic therapy. PMID- 12722392 TI - [Paclitaxel hypersensitivity reactions in patients with advanced ovarian carcinoma]. AB - OBJECTIVES: The purpose of our study was to analyze the rate of hypersensitivity reactions (HSRs) to paclitaxel in patients with advanced ovarian carcinoma. We also analyzed the possibility of re-administration paclitaxel with adequate treatment after hypersensitivity reactions. MATERIALS AND METHODS: The incidence of HSRs was analyzed retrospectively in 112 patients who received 24-hr infusions of paclitaxel (135 mg/m2) for advanced ovarian cancer at the Department of Gynecology Medical University of Gdansk between the January 2000 and the February 2002. Before each course of the paclitaxel administration patients received orally dexamethasone (20 mg p.o., 12 and 6 hours prior to the chemotherapy) and thirty minutes prior to the infusion of paclitaxel all patients received, diphenhydramine (50 mg), and cimetidine (300 mg) intravenously (i.v.). When HSR was observed administration of paclitaxel was temporally stopped and before the re-challenge additional intravenous dosage of hydrocortisone (200-500 mg) and diphenhydramine (25 mg) was given. RESULTS: Severe hypersensitivity reactions occurred in 9 patients (8%) and the administration of paclitaxel was withdrawn. Mild hypersensitivity reactions were observed in 8 patients (7%) and all of these patients were successfully retreated with paclitaxel without HSRs. CONCLUSION: We conclude that this premedication and treatment strategy provides lower hypersensitivity reactions rate and allows for continuing administration of paclitaxel in patients with advanced ovarian carcinoma. PMID- 12722393 TI - [Evolution of surgical treatments in patients with borderline ovarian tumors]. AB - OBJECTIVES: The aim of this study was to evaluate a type and range of primary operation of patients with borderline ovarian tumours. MATERIAL AND METHODS: The analysis included 129 patients operated for ovarian tumours of borderline malignancy in the Gynaecological Department of Medical University of Gdansk between 1978-2000. The two types of primary operations were defined: conservative and radical. The dependency of type of operation on such factors as age of patients, parity, stage and histological type on was assessed. RESULTS: The group of 42 patients were treated with conservative surgery and 87 underwent radical treatment. 36 from 42 patients (85.7%) were treated by USO. A conservative surgery was mainly performed in young patients with mucinous borderline tumor in stage IA. The risk of recurrence was 4.8% in the group of patients treated conservatively and 8.0% in group of patients treated radically. 5-years survival was, 98.2% and 87.2% respectively. CONCLUSIONS: The type of the primary operation depends on age, parity, stage and histological type of borderline ovarian tumors. A conservative surgery is a proper treatment for young women with borderline ovary tumours in stage Ia. For older, perimenopausal women TAH with BSO is suggested. PMID- 12722394 TI - [Evaluation of the effectiveness of second-line intraperitoneal chemotherapy for patients with ovarian cancer]. AB - OBJECTIVES: From a theoretical viewpoint, intraperitoneal therapy in patients with ovarian cancer, a malignancy, which remains mainly confined to the peritoneal cavity is logical. Intraperitoneal catheters have moved to the forefront as a delivery system in cancer treatment. DESIGN: The authors sought to evaluate effects of intraperitoneal chemotherapy (IPC) as a second line therapy for ovarian cancer patients. MATERIAL AND METHODS: From January 1996 to January 2002, 92 patients with recurrent or persistent cancer, after surgery, and first line chemotherapy, were treated with intraperitoneal chemotherapy as a second line treatment. Only 74 were included in the study, due to incomplete of therapy (6 patients), spontaneous fold-out of catheter (3 patients), five patients were treated because of some other kind of carcinomas, three patients passed away during therapy because of independent reasons, and weren't be verified and a patient who had wrong pathological diagnosis in SLL. RESULTS: The three year survival in the whole group reached 58.62% for patients who responded to the first line chemotherapy, or when the debulking surgery was completed, which was a significant improvement in survival. There was a significant improvement in survival for patients with residual tumor < 5 mm compared with the whole group, and especially with these, whose residual tumors were greater then 5 mm. CONCLUSIONS: 1. Survival was increased for patients who had a positive response to the first line intravenous chemotherapy, or had complete a debulking surgery 2. The response for IPC depends on the size of residual disease. 3. Intraperitoneal chemotherapy improves survival in ovarian cancer. PMID- 12722395 TI - [New techniques with biodegradable Valtrac-Bar rings for intestinal anastomosis management of advanced cancer]. AB - AIM: In the study we tried to demonstrate usefulness of the biofragmentable Valtrac-Bar rings in treatment of intestinal obstruction caused by advanced ovarian carcinoma. MATERIALS AND METHODS: Intestinal anastomosis with biofragmentable Valtrac-Bar rings were performed in 26 patients with advanced ovarian cancer with symptoms of intestinal obstruction. In nine patients manifestation of acute or subacute intestinal obstruction symptoms, cachexia and ascites necessitated the use of Valtrac rings during primary surgical operation. In the other 17 women we observed recidive of the disease causing obstruction mainly in the lower part of the digestive tract. Eight sigmoid, six sigmo-rectal and three ileo-ileal Valtrac-Bar rings anastomosis were done after secondary cytoreductive surgery. RESULTS: In 25 treated women we achieved improvement of their general condition, so we were able to continue treatment by chemio- or radiotherapy. Only in one treated patient further relaparotomy with colostomy was needed due to of anastomosis leak. CONCLUSIONS: We conclude that biofragmentable valtrac-Bar rings are very useful, safe and effective tools enabling fast intestinal anastomosis even in patients with inappropriate healing, for instance treated previously by chemio- et radiotherapy. PMID- 12722396 TI - [Use of color Doppler technique and estimation of CA 125 amd TPA levels in blood serum for the diagnosis of adnexal masses]. AB - OBJECTIVE: The purpose of the study was to compare usefulness of color doppler technique and measurements of Ca 125 and TPA levels in differential diagnosis of adnexal masses. MATERIAL AND METHODS: 122 patients admitted and diagnosed due to presence of adnexal masses. Transvaginal scans with color doppler technique were performed Ca 125 and TPA levels in blood serum were measured. Morphological features of tumor during transvaginal scans were also noticed. Histopathological data after operation were collected and compared with preoperative scores. RESULTS: 22 adnexal masses were malignant. Presence of central vascularisation within the tumor and low Resistance Index and Pulsatility Index values were the best predictors of malignancy. Measurements of Ca 125 and TPA levels had worse predictive value ind diagnosis of adnexal masses. CONCLUSIONS: Color doppler technique is a useful tool in diagnosis of adnexal masses and has higher sensitivity and specificity compared with measurements of Ca125, TPA levels and single tumor features achieved during transvaginal ultrasound. PMID- 12722397 TI - [Intraperitoneal hyperthermic perfusion chemotherapy in recurrent ovarian cancer]. AB - The paper presents a treatment of peritoneal spread of ovarian cancer with intraperitoneal hyperthermic perfusion chemotherapy. Hyperthermic antiblastic perfusion was carried out throughout the abdomino-pelvic cavity for 90 min, at the temperature of 41-42 degrees C, with cisplatin (50 mg/l). Perfusion was performed with 4 liters of Maxwell liquid. The perfusion rate was 1200-2000 ml/min. There was no complication during and after procedure. PMID- 12722398 TI - [Own experiences in salvage chemotherapy of ovarian cancer--a preliminary report]. AB - OBJECTIVE: To compare the efficacy and side effects of the ovarian cancer salvage treatment. MATERIALS AND METHODS: 28 patients from 2nd Department of Gynecology of Wroclaw Medical University, treated for recurrent ovarian cancer in stage III and IV were analyzed. First line treatment after complete debulking operation was based on platinum analogue (CAP). Platinum-sensitive patients were administered carboplatin with paclitaxel in the second line treatment. Platinum-resistant cases were treated with paclitaxel, topotecan or liposomal doxorubicin only. RESULTS: The effectiveness of the salvage chemotherapy was similar. Side effects depended on used agent. CONCLUSIONS: We conclude that the results of salvage treatment in ovarian cancer remain unsatisfactory. Introducing new agents into the therapy slightly improved the overall survival time. PMID- 12722399 TI - [Sonoangiography and logistic regression analysis in the preoperative differentiation of ovarian tumors]. AB - OBJECTIVE: To apply logistic regression analysis for several clinical and sonographic data for the construction of a predictive model that could be helpful in the preoperative differentiation of adnexal masses. MATERIALS AND METHODS: Two hundred and eight women with tumors thought to be of adnexal origin were examined preoperatively. Initial analysis included age and menopausal status, ultrasound derived morphological features of adnexal masses (unilateral/bilateral tumors, papillae, septae, tumor size and volume) as well as color Doppler criteria such as PI, RI, Peak Systolic Velocity, PSV assessment. In all examinations we used B&K 2002 ADI (Denmark) and Kretz Voluson V730 (Austria) scanners with transvaginal probes 5-9 MHz. Stepwise logistic regression analysis was used to construct a predictive model that would allow probability of malignancy calculation for individual patient. RESULTS: There were 159 benign and 49 malignant masses. Seven cancers were in FIGO stage one. Statistical analysis revealed that only 5 of initially tested 14 variables had significant influence on the regression equation. These were: age, bilateral mass, presence of septa > 3 mm, papillary projections > 3 mm in the tumor wall and subjective color scale assessment according to Timmerman et al. (1999). Sensitivity and specificity at the 50% probability level of malignancy in the studied tumor were 77.5% and 96.8%, respectively. When 25% cut-off probability level was used, sensitivity increased to 87.7% and specificity dropped to 89.9%. Prospective testing in a new group of 30 patients (5 ovarian cancers) gave sensitivity of 80% and specificity of 100%. CONCLUSIONS: The use of logistic regression analysis can help in modeling clinical and sonographic data. Our model had better predictive value than individual tests and allowed to calculate true probability figure of ovarian malignancy for any given patient with adnexal mass. PMID- 12722400 TI - [Three dimensional sonography and 3D power angiography in differentiation of adnexal tumors]. AB - BACKGROUND: Three-dimensional sonography is a novel diagnostic method proposed to be an additional non-invasive tool in the assessment of ovarian tumors. OBJECTIVE: To study diagnostic potential of 3D sonography and power Doppler angiography in the preoperative differentiation of adnexal masses. MATERIAL AND METHODS: One hundred twenty-eight women with tumors thought to be of adnexal origin were examined preoperatively. Following morphological (papillae, septae, tumor size and volume) and color Doppler (PI, RI, Vmax and TAMX) assessment, 3D ultrasound of adnexal tumors according to Kurjak et al. (2000) was performed. Various scanners were used and included: ATL 5000 HDI (Phillips, USA) and Combison 530 and Voluson 730 (Kretztechnik, Austria) machines. Following variables were studied: inner wall structure, presence of papillae, thickening > 3 mm of septa as well as vascular branching pattern, number and localization of small blood vessels and the presence of vascular anastomoses. RESULTS: Twenty-one tumors were malignant (3 FIGO stage I) and 101 masses were benign. Power Doppler combined with 3D sonography predicted malignancy with a sensitivity of 92.6% (25 of 27 patients). Commonly used morphological and Doppler criteria produced lower sensitivity, the values being in range of 45% to 87.5%. Negative predictive value of 97.2% was the highest for 3D sonography (> 7 points in Kurjak's scale). CONCLUSIONS: Selective use of 3D ultrasound and power Doppler angiography could be used to better characterize adnexal tumors. Detailed 3D sonography may help to identify women who, if needed, may have less invasive surgical procedure such as laparoscopy or be referred to gynecological oncologist. PMID- 12722401 TI - [Gastrointestinal surgery in patients with ovarian cancer]. AB - OBJECTIVES: The objectives were to assess indications as well as outcome and morbidity of gastrointestinal surgery in patients with ovarian cancer. METHODS: The study included 74 patients with primary ovarian cancer who had debulking surgery (bowel surgery) from 1987 to 2001. RESULTS: In our group postoperative residual tumor was--R0 in 15 cases (20.3%) and < or = 2 cm (R2) in 33 patients (44.6%). CONCLUSION: Gastrointestinal surgery is frequently indicated during operation in ovarian cancer. Gynecologic cancer surgeons should be trained accordingly. PMID- 12722402 TI - [Parovarian cysts--not always benign]. AB - OBJECTIVES: The purpose of this study was to determinate the frequency, clinical aspects and management of epithelial parovarian neoplasms among patients hospitalised in our department. MATERIALS AND METHODS: This work has a retrospective character. The research material composed of patients treated in our clinic in the years 1992-2001 because of adnexal mass. In clinical analysis of 13 cases with epithelial parovarian neoplasms we took account of age, symptoms, ultrasound investigations, CA 125 levels, family history, operative treatment, histopathological examinations and follow up. RESULTS: Among 1110 patients operated for of adnexal mass 19.21% constituted parovarian cysts, and 2.6% (13 cases) of them were epithelial neoplasms (11 benign cystadenomas and 2 borderline malignancy neoplasms in FIGO I). The mean age of patient was 42.1. The clinical presentation was lower abdominal pain. CA 125 had the normal range in all patients. In ultrasound investigation parovarian cysts was suspected only in 2 cases when both normal ovaries were apparent. All patients were operated and extension of surgical procedures depended firs of all on age. Two patients with borderline tumors were operated in 1998 and have lived ever since without signs of disease. CONCLUSION: Parovarian cysts are not always benign and broad ligament region may be the point of issue for neoplasms of different histopathological types. PMID- 12722403 TI - [Characteristics of selected features of hereditary ovarian cancer in carriers of constitutional BRCA1 gene mutation]. AB - BACKGROUND: Hereditary ovarian cancer in BRCA1 constitutional mutation carriers shows a characteristic clinical pattern. AIM OF THE STUDY: Evaluation of the type of mutation, age of onset, clinical stage (FIGO) and morphological grade (G) of ovarian cancers in BRCA1 mutation carriers. MATERIAL AND METHOD: We analyzed 16 cases of hereditary ovarian cancers from 14 families. The cases were included into studied group as a result of evaluation of pedigree criteria and molecular genetic analyses detecting constitutional mutation--5382insC, C61G, 4153delA- founder mutations dominating the Polish population. The following features were compared between studied and control group: C1--age of onset of ovarian cancer in 5-year intervals (below 35, 40, 45, 50 and 55 years of age); C2--clinical staging (FIGO) I/II or III/IV: C3--grading (G1-G3). The data were analyzed statistically. RESULTS: Age of onset < 50, OR--2.52 is highly characteristic for BRCA1 mutation carries. Similarly the staging III/IV is more often observed in studied group--OR 2.71. The most characteristic of studied group is high morphological grading--OR 12.38. CONCLUSIONS: 1. Features of HOC in studied material and in other populations are similar 2. High morphological grading is the most characteristic for HOC in BRCA1 carriers. PMID- 12722404 TI - [Evaluation of serum sICAM-1 amd CA-125 in patients with ovarian tumors- preliminary report]. AB - OBJECTIVES: Today's diagnostics and differentiation of ovarian tumour is not efficient enough. Wide spreading tumour immunology of neoplasm presents an opportunity to determine new markers and mechanisms useful for diagnostic and discrimination. The aim of our study was to evaluate concentration of soluble intracellular adhesion molecule-1 (sICAM-1) with CA-125 in ovarian cancer. MATERIAL AND METHOD: Serum of 40 patients undergoing surgical treatment, 32 with ovarian cancer and 8 with benign tumours. Concentration of markers was determined immunoenzymatically. RESULTS: Concentration of sICAM-1 correlated moderately with FIGO stage, tumour volume and CA-125. Mean concentrations between analysed groups were different, but not statistically significant. Determination of sICAM-1 together with CA-125 increased discrimination power of the test. CONCLUSIONS: Determination of sICAM-1 alone seems to have limited application. Addition of this test to CA-125 could improve value of the diagnostic process. PMID- 12722405 TI - [Comparison of quality of life in patients with advanced ovarian cancer treated with intraperitoneal or intravenous cisplatin and cyclophosphamide as a second line of therapy]. AB - OBJECTIVES: The study aimed to compare the effect of second line intravenous and intraperitoneal chemotherapy on physical and psychological aspects of quality of life in patients with advanced ovarian cancer. MATERIALS AND METHODS: Quality of life was measured with EORTC QLQ-C30 (version 3.0) questionnaire. 42 sample patients with histologically confirmed diagnosis of advanced epithelial ovarian cancer treated with second line intravenous or intraperitoneal chemotherapy were included in the study. RESULTS: Higher score of global quality of life and less side-effects of chemotherapy were observed in the group of patients treated with intraperitoneal chemotherapy. In this group constipation and dyspnoea were less common. CONCLUSION: Intraperitoneal chemotherapy has less negative influence on quality of life than intravenous drug delivery. PMID- 12722406 TI - [Evaluation of surgical complications connected with intraperitoneal chemotherapy in ovarian cancer]. AB - OBJECTIVES: From a theoretical viewpoint, the use of intraperitoneal therapy (i.p.) in patients with ovarian cancer (a malignancy, which remains mainly confined to the peritoneal cavity) is logical. Intraperitoneal catheters have moved to the forefront as a delivery system in cancer treatment. Authors described complications during the placement, usage, and evacuation of Tenckhoff catheters. Some of them report much too high number of surgery complications connected with i.p. DESIGN: We report a number of complications connected with insertions, functions, and evacuations of Tenckhoffa catheter. MATERIAL AND METHODS: From January 1996 to January 2002, 92 patients with recurrent or persistent ovarian cancer, after surgery and first line chemotherapy, have had catheter insertion performed, but only 79 have had performed catheter evacuation: because of: not complete therapy (7 patients), three patients died during i.p. therapy, in three cases intraperitoneal catheter has spontaneously fold out. RESULTS: During insertion total number of complications 9(9.78%), 6 bowel incision, 1 bladder incision, 1 hernia of the linea alba, 1 incision of bowel and bladder. During catheter evacuation total number of complications 9 (11.39%), 8 bowel incisions 1 hernia of the linea alba. Complications connected with catheter function: only 8 of 92 (8.70%) required cessation of chemotherapy prior to its expected completion, 2 fistula of the catheter to vagina, 2 fistulas to bowel, in two cases intraperitoneal catheter has spontaneously fold out due to abscess, one after citostatics flow under the skin, one because of abscess in peritoneal cavity, and problems with citostatics inflow one because of subileus. CONCLUSION: The surgical complications occurring during IPC are not dangerous for patients. IPC is valid and safe way of treatment patients with ovarian cancer. The frequency of complications occurring during insertion of Tenckhoff catheter depends on the way of placement. PMID- 12722407 TI - [Evaluation of endometrium during tamoxifen therapy of breast cancer]. AB - Tamoxifen is one of most common drugs for breast cancer therapy. But its weak estrogenic activity in endometrium can be the source of different abnormalities including even endometrial cancer. OBJECTIVES: The aim of this clinical trial was to compare ultrasound scan results, hysteroscopy and pathomorphological findings of tamoxifen treated patients with breast cancer who complained of bleeding. MATERIALS AND METHODS: Analyzed group consisted of 10 patients treated for abnormal endometrium during breast cancer tamoxifen therapy. RESULTS: Among 10 examined women 7 presented no pathological changes, there was 1 case of simple benign hypertrophy and 2 cases of endometrial polyps. CONCLUSIONS: There is no exact correlation between ultrasound scan results and pathomorphological findings in patients treated with tamoxifen for of breast cancer. Transvaginal ultrasound examination is not efficient screening test for endometrial abnormalities during tamoxifen therapy. PMID- 12722408 TI - [Fine-needle biopsy of the breast in the material of Gynecology and Obstetrics Hospital in Walbrzych]. AB - OBJECTIVES: Since the year of 2000 we have done more than 500 fine-needle aspiration biopsies in our hospital. DESIGN: The main cause was a breast lesion interpreted as suspicion of cancer in ultrasonography or mammography and its verification between benign tumor and a cancer. MATERIALS AND METHODS: 508 women with clinical findings of breast tumor were qualified for fine-needle biopsy procedure. RESULTS: The most common diagnosis was a benign cystic tumor found in 183 cases and the second one--fibro-cystic mastopathy (132 cases). Surgical operation of the breast was performed in 32 cases. In 17 we found breast cancer and in 14 the diagnosis was suspicion of breast cancer. CONCLUSIONS: Fine-needle biopsy is valuable for diagnosis of breast cancer and can be easily learned and used by the clinician. It's safe, cost-effective, and accurate technique, which has no counter-indications and causes little discomfort. Results are available in a short time so the decision about the necessity for excisional biopsy or mastectomy is made quickly. PMID- 12722410 TI - [Adjuvant pharmacologic therapy after fine needle aspiration of breast cysts in women]. AB - OBJECTIVES: The most common and generally accepted methods of breast cysts treatment are fine needle aspiration or surgical excision. None of these methods allow to avoid or decrease number of recurrences. Regarding hormonal disturbances as a main reason for breast cysts also conservative treatment should be given. DESIGN: To evaluate whether adjuvant pharmacologic therapy is effective in breast cysts recurrence prevention. MATERIAL AND METHODS: 30 women (35-54 yrs) were treated with fine needle aspiration and afterwards with bromocriptine, estrogen progestogens and progestogens. RESULTS: The authors shows that breast cysts recurrence is approximately 10 fold decreased in group with adjuvant therapy with bromocriptine, estrogen-progestogens and progestogens than in group treated with fine needle aspiration only. PMID- 12722409 TI - [Hormonal abnormalities in women with breast cysts]. AB - OBJECTIVES: Increased estrogen activity and prolactin hypersecretion is a hormonal reason for Benign Breast Diseases. DESIGN: To evaluate hormonal disorders in women with gross breast cysts. MATERIAL AND METHODS: 30 women with gross breast cysts (over 10 mm in diameter) have been investigated. Cytohormonal vaginal swabs and serum levels of prolactin, estradiol and gonadotropins were evaluated. RESULTS: Normal prolactin levels were found in 96.7% of cases, but in 8 women with recurrent breast cysts after repeated FNA-biopsies prolactin secretion stimulated by metoclopramide was significantly increased. Low progesterone activity in all women and low estrogen activity in 65.3% were shown. Absolute high estrogen activity was diagnosed only in 8.7% women with gross breast cysts. CONCLUSIONS: Nearly all patients with gross breast cysts have normal prolactin and low progesterone levels. High estrogen levels appear very seldom and in most cases these patients are hypoestrogenic. PMID- 12722411 TI - [Oncologic gynecology and the Internet]. AB - The strategy of World Wide Web searching for medical sites was presented in this article. The "deep web" and "surface web" resources were searched. The 10 best sites connected with the gynecological oncology, according to authors' opinion, were presented. PMID- 12722412 TI - [Occurrence of the cervix, corpus and ovarian carcinoma in the years 1984-2001 in women hospitalized in the Gynecological-Obstetric Hospital in Walbrzych]. AB - OBJECTIVE: Retrospective analysis the cervix, corpus and ovarian carcinoma in the years 1984-2001. MATERIALS AND METHODS: 1104 women with cervix, corpus and ovarian carcinoma were investigated during the last 18 years. This time was divided conventionally into six-year periods. They include the occurrence, the age of the patients and clinical advanced degree by FIGO in the time of diagnosis of the disease. RESULTS AND CONCLUSIONS: Decrease of the quantity of women cured because of carcinoma of cervix. Considerable increase of quantity of women with carcinoma of corpus. Increase of the number of patients with carcinoma of ovarium. PMID- 12722413 TI - [Perception of neoplasms and prophylactic measures]. AB - OBJECTIVES: Breast cancer is one of the most frequently medical problems of Polish women. DESIGN: The aim of the study was to answer the question, if imagination of neoplasm disease influence the undertaking of prophylactic activity by nurses and midwives. MATERIALS AND METHODS: Study was carried out among 202 female students from the Faculty of Nursing and Health Studies at the University Medical School in Lublin. Study was carried on the basis of the author's questionnaire. RESULT: Nearly half (99, that is 49.0%) respondents associated neoplasm's disease with suffering resulting in death. The majority (146, it means 72.3%) of women said that they had informed others of guidelines on breast self-control. Only 31.7% of respondents wrote that they practise breast self-examination on a regular basis. CONCLUSIONS: Imagination of neoplasm's disease did not influence regularity of self-examination and undertaking of prophylactic activity against breast cancer. PMID- 12722415 TI - [Quality of life in women with cancers of the reproductive organs--humanistic trends in oncology ]. AB - Diagnosis of cancer and oncologic treatment create stress which influences estimation of their situation and physical feeling. Permanent development of medical sciences enables discoveries of more effective ways struggling with illness. However taking advantage of scientific development should be followed by constant concern about patients' quality of life. PMID- 12722414 TI - [Attitude of medical personnel regarding prophylaxis of neoplasms of the reproductive organs and the breast]. AB - OBJECTIVES: Breast cancer is one of the most frequent medical problems of women in Poland. DESIGN: The aim of the study was to verify, whether and how far nurses and midwives keep fundamental rules of prophylaxis of breast cancer and that of reproductive organs and whether the fact of taking care of a patient with cancer influence their health's behaviour. MATERIALS AND METHODS: Questionnaire study was carried out among 202 people. RESULTS: Only 17.8% of all respondents have not taken any actions to protect themselves against tumours disease. The others tried to assume a healthy life style and took advantage of prophylactic examinations. The variables accepted in the study did not differentiate the surveyed behaviour. CONCLUSIONS: Majority of nurses and midwives have taken prophylactic activity of breast cancer. The fact of taking care of a patient with cancer did not influence their health's behaviour. PMID- 12722416 TI - [Neoplastic cell death induced by photodynamic therapy]. AB - The photodynamic therapy (PDT) is a new therapeutic method aimed at selective destruction of neoplastic cells. In PDT photosensitizing pigments are used, introduced into cells and then activated with laser light. This leads to formation of free radicals in the cell (peroxide aminoradical, hydroxyl radical), or singlet oxygen. The outcome is death of neoplastic cells through apoptosis or necrosis. The effect of PDT on the type of cell death depends on the biochemical properties of the photosensitizer used, and exposure time of neoplastic cells to light. The studies on application of PDT are primarily aimed at directing neoplastic cells towards apoptosis. PMID- 12722418 TI - [From the scientific heritage of V.V. Parin (to the development of problems of space medicine and physiology)]. PMID- 12722417 TI - [Lymph node mapping and sentinel node detection in gynecological oncology]. AB - The aim of this paper is the presentation of the latest opinions on the lymph node mapping and the sentinel node localization in the female genital organ neoplasm. The current strategies of lymph node resection in gynecologic oncology have been presented. The methods of lymph node staining and detection has been expounded as well. The paper also contains the results of sentinel node localization in the vulvar, cervical and endometrial cancers. PMID- 12722419 TI - [Effects of centrifugation-related artificial gravity on the human organism. Positive and negative effects]. AB - Data of investigations of the artificial gravity issues in centrifuge experiments in Russian, USA and Japan over the past 40 years were analyzed. Periodic +Gz centrifugation was used to prevent or eliminate body deconditioning after 3- to 56-d simulated microgravity, and to increase the body functional reserve (orthostatic stability) in ordinary life. Positive effects included prevention of a decline of gravitational stability and reduction or averting the negative effects of simulated microgravity on various physiological systems. Better results can be achieved by combining artificial g-loads with the well-proven methods of combating deconditioning, i.e. physical training on bicycle ergometer and induced hydration. Human tolerance of g-loads on a short-arm centrifuge with a high (100%) gravitational gradient was found no worse than on a mid-arm centrifuge with a lower (20%) gravitational gradient. Negative effects of +Gz on a short-arm centrifuge were revealed and counteracting measures were determined. PMID- 12722420 TI - [Electromagnetic fields: is there any probability of the risk of cancer?]. AB - The literature on epidemiological studies of impacts of humans of electromagnetic fields (EMF) including weak ones has been reviewed. The risk of cancer to people in EMF appears to be a complicated problem. Consideration is given to the phenomenon of radiophobia associated with exposure in EMF. PMID- 12722421 TI - [Correction of the human body hydration in different periods of space flight]. AB - Hydration level of the human body at the end of space flight is not same as at its beginning. This was the reason for development and testing of opposite in action methods for hydration improvement: at the onset of microgravity a dehydration therapy is applied and, on the contrary, in the final period of space flight methods for retaining body fluids are of preference. Consumption of a diuretic and a water-salt supplement by orbiting crews reached the required effect suggesting applicability of the pharmaceutical correction as a measure against dehydration. PMID- 12722422 TI - [Problems of diagnostics and prophylaxis in a Martian flight]. AB - In future interplanetary missions that will take up to two years the role of monitoring of the main human body systems and prevention of the effects of microgravity for the benefit of crew health and performance will rise eminently. In contrast to the orbital flights, diagnostics and prophylaxis will be a responsibility of the doctor within the crew who will consult solely own expertise and judgment. Hence, criteria and requirements to health monitoring and medicare systems and items are to be defined. The paper substantiates a concept and principles of diagnostics and prophylaxis, ideas concerning the scope of associated measures on the way to Mars, during the visit to the planet and on return to Earth; potentiality of a short-arch centrifuge as a means of prophylaxis is also discussed. PMID- 12722423 TI - [Hormonal regulation of metabolism in the human body in microgravity and during simulation of its physiological effects]. AB - The paper presents results of investigations into the effects of space flight and simulation experiments of various length on the hormonal regulation of metabolism in the human body. Microgravity was shown to instigate shifts on different levels of the hormonal regulation and consequent adjustment of metabolism to this new environment. For instance, adaptation occurs on the level of basal secretory activity resulting in altered metabolism and formation of a pool of hormones. Metabolism readaptation to the Earth's gravity is dependent on polymorphic processes in the system of hormonal regulation developing in the course of time. Trends in the hormonal regulation of water-electrolyte metabolism during early adaptation point to inequality of contributions of the antidiuretic hormone, natriuretic peptide, and the renin-angiotensin-aldosterone system. In the ground based simulations responses of the hormonal regulation of water-electrolyte metabolism differ in intensity and types of hormones involved. Temperature variation can modify reactions of the comosis and volume regulating hormones at the beginning of adaptation. Physical-chemical regulation of calcium homeostasis in microgravity reveals itself by a rapid decline of the calcium-binding ability of blood buffers and, later on, degradation of the relative ability of extraplasmic structures to bind calcium. Qualitative and quantitative changes in the diurnal rhythm of the suprarenal steroidogenesis are indicative of modification of intensity of reactions of the main biosynthetic sequences. Countermeasures used by test-subjects in these investigations loosened significantly the aldosterone-secreting biosynthetic sequences but were favorable to the synthesis of testosterone and hydrocortisone. Some of the highly variable processes of hormonal regulation were mute to the diurnal rhythms in the pre flight and preexperimental periods. PMID- 12722424 TI - [Dynamics of some electrocardiographic parameters in cosmonauts during long-term Mir mission]. AB - The paper deals with the results of electrocardiographic investigations of relatively resting 59 cosmonauts at the age of 30 to 52 during long-term space missions. Heart rate (HR), T-wave amplitude and an electric systole interval were analyzed. During their missions, the cosmonauts consistently combined HR rises with aggravated repolarization manifested by a relative elongation of the phase and T-depression. All alterations were statistically significant (p < 0.05); however, no pathology was detected in the myocardial bioelectrical activity. With this dynamics, values of the parameters under study did not, as a rule, leave the boundaries of the physiological norm. PMID- 12722425 TI - [Prophylaxis of the unfavorable shifts in mineral turnover in bones during extended exposure to the conditions reproducing the physiological effects of microgravity]. AB - Effectiveness of the countermeasures and methods for mitigation of the unfavorable effects of microgravity on mineral turnover and properties of bones was evaluated in simulation experiments with animals (penal immobilization, suspension) and with participation of human subjects (37 to 120-d head-down bed rest). The experiments with rats were aimed at testing pharmaceutical preparations (bisphosphonates), determining doses, plan and way of administration. Preparations of this group differ considerably in their action on osteogenesis, mostly because of specifics of the chemical structure. Nonetheless, optimization of the course of treatment masked significantly side effects without detriment to the preventive action. Head-down bed rest was used to test effectiveness of physical exercises (PE) as a main countermeasure against the microgravity impacts. Both standard and experimental PE regimens were tested. Several groups of test-subjects combined PE with consumption of xidifon (a bisphosphonate). In addition, mineral supplements (potassium and calcium) were used. The PE + xidifon combination and regulation of mineral intake with foods was found to reduce calcium losses during 370-d bed rest and prevent a material degradation of strength of bone samples, and essentially rehabilitate shifts in the hormonal regulation of calcium turnover and calciuretic function of the kidney. PMID- 12722427 TI - [The syndrome of visual asthenopia in ground aviation personnel: prevention, correction, rehabilitation]. AB - A complex of measures for prevention, correction and rehabilitation of ground aviation personnel revealing symptoms of vision fatigue due to the work with electronic information display systems is proposed. Yet, the author point to the need to apply these measures as a part of an integral approach. PMID- 12722426 TI - [Respiration and cardiovascular mechanisms of hypobaric correction of the head down redistribution of local blood volumes]. AB - The focus was placed on the physiological mechanisms of correction of the blood redistribution during the head-down tilt (HDT) with a discrete or complex use of the methods of negative pressure respiration (NPR) and lower body negative pressure (LBNP). It was evidenced that rise in the intracranial hydrostatic pressure in HDT (-6 degrees) can be compensated by NPR within a range of -10 to 15 cm of water column causing decrease in the intrasternal pressure from -5.04 to -7.74 cm w.c. Bioimpedance and ultrasonic investigations led to the conclusion that the decreased transpulmonary pressure is responsible for blood outflow from the intracranial venous system till the level adequate to the horizontal position of the body. The results were verified in experiments with unconscious cats: pressure in the exterior jugular vein and v. cava superior during HDT (-30 degrees) with NRP at -5 cm w.c. decreased by 3.2. and 4.3. w.c., respectively. Experiments with human subjects also demonstrated that the complex use of NPR and LBNP produces an additive hemodynamic effect and can be considered a new method for correcting the adverse consequences of redistribution of the local blood volumes toward the head end. PMID- 12722428 TI - [Biomass and net primary productivity of artificial tropical rainforest in Xishuangbanna]. AB - The stand biomass and primary net productivity of artificial tropical rainforest in Xishuangbanna were estimated, based on sample tree method and harvesting method. The results showed that the standing biomass was 390.4 t.hm-2, of which, 362.5 t.hm-2(92.8%) were contributed by tree layers. The biomass of shrub and inter-layer plants (including epiphytes) was 19.3 t.hm-2(4.9%) and 3.6 t.hm 2(0.9%), respectively, and that of herbaceous layers was 5.0 t.hm-2. The primary net productivity of the stand was 2227.3 g.m-2.yr-1, of which, 1553.5 g.m-2.yr 1(69.7%) were contributed by tree layers. In the allocation of primary net productivity in different parts of trees stems showed the highest net productivity, accounted for 42.0%. Leaves and branches were accounted for 30.2% and 13.5%, respectively. The leaf area index (LAI) was 7.061. The optimum regression models of different dominant plants and organs of the sample trees of tree layer in the artificial tropical forest were built. PMID- 12722429 TI - [A comparison study on the species diversity between the gap and non-gap in Jinyun Mountain]. AB - The species diversity and community stability of gap and non-gap stands in Jinyun Mountain were studied by using species richness index(dGL), Shannon-Weiner index, Pielou evenness index(J), Simpson index(D), ecological dominance(lambda), evenness-dominance-abundance index(Z) and community dominance(C). The results showed that the dGL, H', J and D calculated by individuals numbers in gap are 12.14, 4.62, 0.70, 13.32, respectively. Shrub layer plays a greater role than other layers. The corresponding indices in non-gap stand are 6.32, 3.74, 0.66 and 8.16, respectively, which were lower than those in gap. Species diversity indices of the community were far higher in gap than in non-gap. However, ecological dominance and community dominance in gap were smaller than those in non-gap significantly. It suggested that species diversity of the community increased, and community stability decreased, due to the existence of gap formed by natural or human disturbance. PMID- 12722430 TI - [Fractal characteristics of particle size distributions of mangroves soils in Yingluo Bay]. AB - Based on fractal theory, the fractal characteristics of particle-size distributions of mangrove soils in Yingluo Bay (21 degrees 28'N, 109 degrees 43'E) were studied. The results show that the fractal dimensions of the soils ranged from 2.6837 to 2.8834, and decreased in the order of sand loam < light loam < medium loam < heavy loam < light clay. The fractal dimensions of the soils on exterior beach were lower than those on middle and inner beaches. There was a significant positive linear relationship between fractal dimension and soil salinity and organic matter content. The major factors that influenced the fractal dimensions of the soils were community type, soil texture, beach position, soil salinity, and soil organic matter content. PMID- 12722431 TI - [Characteristics of soil water infiltration in sub-alpine dark coniferous ecosystem of upper reaches of Yangtze River]. AB - Dark coniferous forest is the predominant type of vegetation in the upper reaches of Yangtze River. Difference among different types of soil exists. The sand content of soil is higher and the soil texture is coarser in the early stage of forest succession. The sand content of soil decreases with the advancement of the forest succession while that of soil in Abies fabri over-mature forest is the lowest. In slope wash soil, the sand content of soil decreases with the increasing soil depth. The soil porosity and soil water-holding capacity increases and soil bulk density decreases with the advancement of forest succession and decrease of soil depth. The deeper soil depth or the smaller soil water content are, the smaller the unsaturated hydraulic conductivity of soil measured by CGA method. Moreover, the correlation of soil water content with unsaturated hydraulic conductivity of soil can be simulated by an exponential function. The saturated hydraulic conductivity of soil decreases exponentially with the increasing soil depth. The time to attain the stable infiltration rate is different among different soil depth, while the deeper the soil depth is, the longer the time needs. The variation in soil texture, soil physical properties and the high infiltration rate of soil there implicated that there are scarce surface runoff, but abundant in subsurface flow, return flow and seepage, which is the result of regulation by dark coniferous forest on hydrological processes. PMID- 12722432 TI - [Structural characteristics of Abies fabri forests at the upper reach of Yangtze River]. AB - The structural characteristics of Abies fabri forest under different succession stages in Gongga Mountains at the upper reach of Yangtze River were studied. The results showed that in the mature Abies fabri forest, there existed abundant seeds and a few saplings of Abies fabri younger than 20 years, but only Abies fabri was the dominant regeneration species. In the poplar-fir mixed forest, the height growth of poplar and birch was very fast during first 30 years, and poplar and birch dominated rapidly over the canopy. Abies fabri had a lower growth rate and a strong shade-tolerance in its first growth stage, and could replace other tree species gradually. The cycle of mud-rock flow occurrence was above 100 years in the Gongga montane areas from elevation of 2800 to 3200 m. After mud-rock flow, poplar and birch often occurred and dominated, and there were only a small number of Abies fabri saplings in slash. Under natural condition, to recover Abies fabri forest would demand a long time, but this process could be controlled and improved by human activities. PMID- 12722433 TI - [Niche characteristics of dominant populations in natural forest in north Guangdong]. AB - Two communities of the natural evergreen forest in Luoba Nature Reserve, North Guangdong, each represented by a 2500 m2 plot, were surveyed with Tree-layer frequency method. Based on these data, the niche characteristics of 12 dominant populations, i.e., Castanopsis fargesii, Pinus massoniana, Schima superba, Liquidambar formosana, Diospyros morrisiana, Myrica rubra, Castanopsis lamontii, Adinandra milletii, Castanopsis eyrei, Elaeocarpus silvestris, Daphniphyllum calycinum, Neolitsea chuii, were described and analyzed, with stand layer as the resource state and tree abundance as the resource state descriptor of niche. The results indicate that most of the shade-tolerant tree species have higher niche breadth values, while the intolerant tree species have the lower ones, with zero value for Pinus massoniana and Liquidambar fomosana, which is in accordance with the declining status of the two species in the community. All the dominant populations, both shade-tolerant and intolerant, showed some adaptation to the community environment. 54.5% of the population-links had a niche similarity over 0.5, and 51.5% of them had a niche overlap over 0.2 in community 1. Niche characterization combined with conventional Tree-layer-frequency method could better determine the status and regenerating potential of tree species in the community. On the other hand, measuring tree abundance in different stand layers as the resource niche could demonstrate tree's requirement for the habitat. Variations in tree composition result in varied niche breadth, which lead to the variations of niche similarity and niche overlap values. PMID- 12722434 TI - [Biomass allocation and growth analysis on the ramets of Phragmites communis populations in different habitats in the Songnen Plains of China]. AB - In the Songnen Plains of China, the reeds grow fastest in middle August. In this experiment, four research plots were chosen in the arid habitats. The first plot was dominated by reed, and in which, there was some stagnant water in July and August. The ramets of reeds grew singly, and most of them could turn into reproductive growth. Its community coverage was more than 95% after the fast growing season of July. The second plot was Leymus chinensis + Phragmites communis community, where soil was alkaline meadow with stagnant water for a short time or not. Its community coverage was more than 90%. Two or three ramets grew in a tuft, while most of them couldn't turn to reproductive growth, so reeds were used to make hay stored in winter every year. The third plot was on Aeolian sandy soil with good permeability and drainage on a grassland of artificial forest edge on sand dune. Its community coverage was about 50%, and two or three ramets grew in a tuft. The last plot was alkaline bareground patches, where top soil was lost completely. Soil was hard and permeability was poor. On some alkaline patches, there could form a micro-community dominated by reed, and eight to twenty ramets could grow in a tuft. Ramets were all slender and creeping stems. Its community coverage was from 20% to 40%. The results showed that four mentioned habitats, the growth and productivity of the ramets of the populations were the largest in the lowland and the smallest in the alkaline patch. The height of ramets differed 2.8 times, and the biomass differed 4.4 times in the two habitats. There were also significant difference between the other two habitats, sand soil and mixed-grass community. The integrated condition of habitat were the major factor that affected the height of the ramets of the population. Moreover, the growth condition of interior habitat was the major factor that affected the ramet biomass of the population in lowland and alkaline patch, while the integrated habitat conditions had much more effects on the ramet biomass of the population in sand land and meadow. The height of ramet was much lower in ecological plasticity than the biomass in every population. The variations of biomass allocation of the ramet in different habitat conditions indicated important strategies in the growth regulation and biomass allocation. More biomass of ramets was allocated to leaves preferentially in the four habitants, especially on sand dune. Reeds in alkaline patch had small ramets and enough growth spaces. There were 63.04% and 53.61% biomass allocated to leaves, and only 14.82% and 19.92% to the stems, although 58.31% was allocated to leaves and only 19.05% to stems to ensure the full material product in the meadow, because of the interspecific competition. But, in the lowland with better conditions where had a higher density and large coverage, only 42.1% were allocated to leaves, while 37.52% were allocated to stem to elongation of stem in order to improve the individual competition for space and light. The leaf sheath/biomass ratio of the ramets was 20.38%-26.47%, which was nearly, at the same level in four habitats. There was the same regulation of the heterogeneous speed growth by power function, either in the increased height and weight of the ramets, or in the relative increased weight of the stems, leaves and the ramets of the populations in four different habitat conditions. PMID- 12722435 TI - Relationship between resistance and growth of Trifolium repens plants and their disease history. AB - 17 healthy host clones and 14 clones originally diseased with the annual rust, Uromyces trifolii-repentis, were sampled from the natural population of Trifolium repens, which were used as the experimental plant materials. 10 rust strains of Uromyces trifolii-repentis were collected for the experimental inoculation. Two greenhouse and two field experiments were done, with three treatments as inoculated with one rust strain, inoculated with ten rust strains and controls, respectively. The results showed that originally healthy plants were more resistant, and originally diseased ones were more susceptible to the rust. However, there was a considerable overlap among some of the clones. Pot experiments, both in the greenhouse and in the field, revealed no significant differences in growth between originally healthy and originally diseased plants within the treatments (controls, inoculations with one strain and with ten strains). However, there were significant differences in growth between treatments. This lack of difference in growth within treatments contrasted to the result of a transplantation experiment in a natural meadow where the rust was absent. The transplantation experiment showed that originally diseased clones produced significantly more leaves than originally healthy ones (mean + S.E.: + 0.77 and 17.6 + 0.49, respectively). Further, originally diseased clones showed more variation in leaf production, compared with the originally healthy ones. The difference between the pot experiments and the transplatation experiments tells that to study individual differentiation in responses to environmental factors, such experiments ought to be performed under a range of natural conditions, where various biotic and abiotic factors are involved. PMID- 12722436 TI - [Relationship between modular growth characteristics of five plant species and water and heat combination in warm-temperate and moderate-temperate steppes]. AB - Applying comparative study on the characteristics of modular growth of five mutual species such as Stipa grandis, Leymus chinensis, Cleistogenes squorrosa, Lespedeza dahurica and Heteropappus altaicus, the grey-correlative analyses between the demographic growth analysis indexes and average temperature, precipitation and moisture had been carried out during months of the growth seasons from 1999 to 2000 in Research Stations of warm-temperate and moderate temperate steppes of Inner Mongolia. The results demonstrated that the indexes of DRGR & DRGRa and DULA of these species were all higher in moderate-temperate steppe than warm-temperate steppe, showing the ecological characteristics of adaption themselves to the climate of shorter growth season and inadequate thermal condition, while longer duration of leaf area and modular quantity (DLAD & DMND) of these species reflected the adaption of the longer growth season and sufficient quantity of heat in warm-temperate steppe. Meanwhile, the greycorrelative degree between modular growth indexes of these species and moisture was remarkably higher in warm-temperate steppe than in moderate temperate steppe, showing that modular growth of plant individual was more sensitive to the combination of water and heat in growth season in warm-temperate steppe, and indicating the response characteristics of plant modular growth in different thermal steppe region. PMID- 12722437 TI - [Protective mechanism in photoinhibition of photosynthesis in Citrus unshiu leaves]. AB - By using a portable PAM-2000 fluorometer and a spectrophotometer (722 Model), the diurnal variation in main chlorophyll fluorescence parameters such as initial fluorescence (F0), maximal fluorescence (Fm), ratio of variable fluorescence to maximal fluorescence (Fv/Fm), non-photochemical quenching (qN), fast-phase (qNf) and slow-phase (qNs) of non-photochemical quenching, electron transport rate (ETR), and relative content of xeaxanthin in Citrus unshiu leaves were observed on fine days. The results showed that ETR, qN, qNf, qNs and relative content of xeaxanthin increased with increasing light intensity, but Fm, F0, photo-chemical efficiency of PS II (Fv/Fm) decreased. After treated by DTT, qNs was much lower, but F0 was much higher than control. These results indicated that xanthophyll cycle and delta pH existed, and played an important protective role against photo damage to the photosynthetic apparatus in Citrus unshiu leaves during diurnal variation of photosynthetic efficiency. PMID- 12722438 TI - [Ecophysiological effects of multiple cropping of winter wheat-spring corn-summer corn in Huanghuaihai Plain]. AB - Compared to sequential cropping of winter wheat-summer corn and mono cropping of spring corn, the ecophysiological effects of multiple cropping of winter wheat spring corn-summer corn in Huanghuaihai Plain were studied. The results showed that under the multi-cropping, the crops occupied higher spatial niches during the period of reproductive growth. Ecological factors such as light, temperature, and air were improved, and plane light acceptance was changed into multistory light acceptance, which made the relative intensity of illumination in crop communities increased. Moreover, soil temperature between rows and wind velocity in planting strips were also increased. All these changes were advantageous to increasing the intensity and velocity of grain filling. The chlorophyll content and photosynthetic rate in functional leaves of crops were higher, which was the main reason of yield increase under multiple cropping. PMID- 12722439 TI - [Thermo-sensitive period and critical temperature of fertility transition of thermo-photo-sensitive genic male sterile wheat]. AB - The thermo-sensitive period and the critical temperature of fertility transition of C49S, a principal thermo-photosensitive genic male sterile line in two-line hybrid wheat, was studied in the growth chambers for controlling temperature and photoperiod. The seeds were sown on different time for some years. The results showed that the thermo-sensitive period in fertility expression of C49S was from PMC formation stage to mature pollen stage, and there were two most sensitive stages to temperature on fertility expression. One was the PMC meiosis stage, and the other was the middle microspore stage. The critical temperatures evoking a complete male sterility were the mean minimum temperature at PMC meiosis stage (Tmin1), the mean temperature at microspore stage (T2) and the mean minimum temperature at microspore stage (Tmin2) lower than 8.5 degrees C, 13.5 degrees C and 10.5 degrees C, respectively. The critical temperatures keeping a nearly normal male fertility Tmin1 and T2 and Tmin2 were higher than 11.5 degrees C, 15.0 degrees C and 12.5 degrees C, respectively. The value as well as the conditions and the risks of thermo-photo-sensitive genic male sterile line of wheat applied to hybrid wheat were evaluated in this paper. PMID- 12722441 TI - [Effects of forms and application rate of nitrogen fertilizer on yield and qualities of tobacco in southeast Tibet]. AB - Field experiments in southeast Tibet were conducted to study the effect of nitrogenous fertilizer supply and its forms on tobacco. The result indicated that the yield, production value, and the contents of nicotine, potassium oxide and total nitrogen of the cured leaves were positively correlated to the amount of N application, and significantly and negatively correlated to the percentage of superior-medium class leaves of tobacco and contents of deoxidize sugar. The highest yield and best quality of tobacco were obtained by applying 75 kg N per hectare. Inorganic nitrogenous fertilizer could significantly improve the yield and quality of tobacco, compared with organic nitrogenous fertilizer. Although there was no significant difference among ammonium N, nitrate N and nitrate ammonium N application, the effects of ammonium N, nitrate N, and nitrate ammoniun N application were significantly higher than that of urea application. The effect of ammoniun N application on tobacco yield and quality was better than that of nitrate N application, and the important reason was the great difference in the process of absorption and assimilation of ammoniun N and nitrate N. PMID- 12722440 TI - [Physiological adaptability of seeding rice genotypes with different P uptake efficiency under low P-deficient stress]. AB - The genotypic differences in P uptake efficiency of three rice varieties (IR74, IR71331 and IR71379) were studied under the P-deficiency condition with hydroponics, and their adaptability to low phosphorus stress about physio biochemical mechanisms was further studied. The results showed that rice genotypes tolerated low P stress resulted from the co-ordination of P uptake efficiency, internal utilization efficiency and its translocation efficiency. The higher P-efficiency type IR74 and the middle type IR71331 had a higher P uptake efficiency. The rice genotype with higher P-efficiency was characterized by higher activity and desirable kinetic parameters of H2 PO4- uptake, showing lower Km and Cmin, but higher Imax values and relative APase activity, small amount of Km and Cmin. Moreover, under low P stress, the activity of RNase was about ten to fifteen times as high as that of the control (CK), but it had little genotypic differences. PMID- 12722442 TI - [Cycling and balance of copper in farmland ecosystem of Jianghan plain in Hubei Province]. AB - The cycling and balance of copper in 8 cropping systems, i.e., wheat-rice, rice rice, rape-soybean, rape-peanut, wheat-sesame, wheat-cotton, pepper-cabbage and radish-eggplant in Jianghan plain of Hubei Province were studied. The results showed that the main output of copper was crop harvesting, up to 85.9%-95.1% of total output, and the output of copper by field runoff and soil leaching was low. The main inputs of copper were organic fertilizer, rainfall, falldrop, and natural return. The inputs of copper from inorganic fertilizer and seeds (seedlings) were low. The balance analysis pointed out that copper had an accumulative trend in two patterns of rice and commercial vegetable ecosystem, and was in balance in upland field ecosystem. PMID- 12722443 TI - [Effect of long-term fertilization on NO3(-)-N accumulation and moisture distribution in soil profiles]. AB - On the basis of long-term fertilization experiments in gully area of Loess Plateau, the relationships among fertilizations, NO3(-)-N accumulation, and moisture distribution in soil profiles were evaluated. The results showed that fertilization and rainfall significantly influenced the yields. NO3(-)-was accumulated in soil profiles under N and P fertilizers and organic manure(NPM) and N and P fertilizer (NP) treatment. The accumulated NO3-N reached 340 kg.hm-2 in 60-120 cm under NPM treatment, and 220 kg.hm-2 in 80-140 cm under NP treatment, respectively. Soil moisture in 100-300 cm was significantly reduced under NPM treatment. Under NP treatment, it was significantly reduced in dry year and normal year, while under organic manure(M) treatment, it was significantly reduced in dry year. Soil moisture under P and CK treatments was relatively stable in different years. N uptake under NPM and NP treatments was significantly decreased in dry year. It reflected the close relationship among soil moisture, crops, and fertilization. PMID- 12722444 TI - [Degradation of metsulfuron-methyl in soils. I. Effect of soil properties]. AB - The ANOVA results showed that degradation of metsulfuron-methyl was different among various soils. Five principal factors were selected by principal factor analysis and they represented 90.4% of total information. The multivariate regression equations were established between soil properties and metsulfuron methyl residues after stepwise regression analysis. The partial correlation and path analysis indicated that soil pH and microbial activity were the most significant factors controlling the degradation of metsulfuron-methyl in soils. PMID- 12722445 TI - [Analysis on landscape pattern in the Tuoketuo region of Inner Mongolia]. AB - The Tuoketuo region is located in the middle reach of Yellow River, farming pastoral zone in Inner Mongolia. In this paper, the vegetation map of study area was compiled by combining TM image and field vegetation survey. Applying several landscape pattern indices such patch size, number, perimeter density, diversity index, evenness index and fragmentation index etc., the landscape pattern of the Tuoketuo region was analyzed on a GIS platform. The result showed that the landscape types in Tuoketuo region mainly consisted of farmland, artificial forest and grassland. The area of farmland was 281.72 km2 and accounted for 33.64% of study region, artificial forest was 22.0%, and grassland was 6.65%. The landscape matrix in northeast part of study area was farmland. On the farmland matrix, Populus + Ulmus patches distributed with centering of residential spot, there were 89 patches. The patch density was 2.568 patch.km-2, and the mean patch area of Populus + Ulmus was only 0.39 km2, therefore, the characteristics of matrix and patch was very typical. Perimeter density of Populus + Ulmus and hill gully were higher than other landscape types, indicating that the fragmentation degree of the former was higher, and hill-gulf was cut seriously in study area. All of these results emphasize that human activity had a strong impact on the landscape pattern in the Tuoketuo region. PMID- 12722446 TI - [Application of ordinary Kriging method in entomologic ecology]. AB - Geostatistics is a statistic method based on regional variables and using the tool of variogram to analyze the spatial structure and the patterns of organism. In simulating the variogram within a great range, though optimal simulation cannot be obtained, the simulation method of a dialogue between human and computer can be used to optimize the parameters of the spherical models. In this paper, the method mentioned above and the weighted polynomial regression were utilized to simulate the one-step spherical model, the two-step spherical model and linear function model, and the available nearby samples were used to draw on the ordinary Kriging procedure, which provided a best linear unbiased estimate of the constraint of the unbiased estimation. The sum of square deviation between the estimating and measuring values of varying theory models were figured out, and the relative graphs were shown. It was showed that the simulation based on the two-step spherical model was the best simulation, and the one-step spherical model was better than the linear function model. PMID- 12722447 TI - [Chemical components of volatile oil from Mikania micrantha and its biological activity on insects]. AB - The chemical components of volatile oil from Mikania micrantha were analyzed by GC/MS. Twenty-two compounds were identified, and monoterpenes and sesquiterpenes as well as alcohol and ketone with their derivatives were major components. Biological activity of volatile oil on insect was investigated. The results showed that the volatile oil had significant deterrent effect on oviposition of Plutella xylostella, Phyllotreta striolata and Phaedon brassicae at dose of 5-10 microliters.plant-1. They also possessed established contracting toxicity, and their rectifying reduce rates for Lipaphis erysimi were 50.0%, 59.86%, 62.51% at concentration of 500, 750, and 1000 mg.L-1, respectively. However, they had not fumigating toxicity for virginogeniae, L. erysimi, 2nd instar larvae of P. xylostella, and adult of P. striolata at concentration of 500 mg.L-1. PMID- 12722448 TI - [Suppressive effect of secondary substances on Liriomyza sativae population]. AB - Life table and interference index of population control (IIPC) were used to evaluate the effects of spraying alcohol extracts of plant secondary substances on the population of Liriomyza sativae. Field experiment results showed that alcohol extracts derived from Lantana camara, Eupatorium odoratum, and Wedelia chinensis, and one kind of commercial petroleum spray oil were positive to protect kidney bean from infestation of L. sativae. The IIPC of these repelling substance compared with control were 0.136, 0.163, 0.213, and 0.222, and the control effects on L. sativae were 86.4%, 83.7%, 78.7%, and 77.8%, respectively. At the same time, the parasitism of vegetable leafminer was increased compared with control, indicated that the effects of parasitism were increased. The results indicated that these substances had strong effects in controlling the population of L. sativae. PMID- 12722449 TI - [Using chlorella and effective microorganisms to optimize aquatic ecological structure and to regulate water quality]. AB - To optimize aquatic ecological structure and to regulate water quality, Chlorella vulgaris and effective microorganism were added to Exopalaemon carinicauda pond and fishponds. The results showed that after adding Chlorella vulgaris to the shrimp pond and fishpond, Chlorella vulgaris turned into a dominant species, and its amount was 16.92 and 4.76 times of CK. The zooplankton biomass reached to 4.32 mg.L-1 and 2.84 mg.L-1, increasing by 19.3% and 2.5%, compared with CK, respectively. Rhodospirillaceae, photosynthetic bacteria and yeast saccharomycete in the ponds could obviously change the composition, number, ratio, and biomass of the plankton (phytoplankton and zooplankton), and adjust aquatic chemical environment. The treatment of "Saccharomycete + Nitrifying bacteria" decreased the concentrations of NH4+ obviously, which was only 44% of CK. The BOD and COD in shrimp ponds were only 56.5% and 38.4% of CK. The treatment could increase the dissolved oxygen and primary production in the pond. PMID- 12722450 TI - [Toxicity identification evaluation on efficiency of chemical effluent treatment]. AB - Acute toxicity tests of the effluents from both inlet and outlet of the treatment system in a chemical plant in Jiangsu Province were conducted with Daphnia magna. The results showed that both effluents were toxic to this species. Toxicity identification evaluation (TIE) was used to identify the key toxicants in the effluent before and after treatment. The results indicated that Cu2+ was the key toxicant causing the toxicity of input water, and some metals and polar organics were the coexistent toxicants in the water. The toxicity of output water was caused by some toxic organics, which were easy to be oxidized under acidic conditions. The results also showed that the toxicity removal efficiency reached 77.6% through the treatment process. It could be seen that the effluent treatment process in this plant had a higher efficiency of removing heavy metals, but the removal rate of organic compounds was quite low. It was concluded that the production techniques and the effluent treatment process of this plant were not perfect, and needed to be improved. PMID- 12722451 TI - [Relationship among soil enzyme activities, vegetation state, and soil chemical properties of coal cinder yard]. AB - From field investigation and laboratory analysis, the relationships among soil enzyme activities, vegetation state and soil chemical properties of coal cinder yard in thermal power station were studied. The results showed that vegetation on coal cinder yard was distributed in scattered patch mainly with single species of plant, and herbs were the dominant species. At the same time, the activity of three soil enzymes had a stronger relativity to environment conditions, such as vegetation state and soil chemical properties. The sensitivity of three soil enzymes to environmental stress was in order of urease > sucrase > catalase. The relativity of three soil enzymes to environmental factor was in order of sucrase > urease > catalase. Because of urease being the most susceptible enzyme to environmental conditions, and it was marked or utmost marked interrelated with vegetation state and soil chemical properties, urease activity could be used as an indicator for the reclamation of wasteland. PMID- 12722452 TI - [Advance in researches on the effect of forest on hydrological process]. AB - According to the effects of forest on hydrological process, forest hydrology can be divided into three related aspects: experimental research on the effects of forest changing on hydrological process quantity and water quality; mechanism study on the effects of forest changing on hydrological cycle, and establishing and exploitating physical-based distributed forest hydrological model for resource management and engineering construction. Orientation experiment research can not only support the first-hand data for forest hydrological model, but also make clear the precipitation-runoff mechanisms. Research on runoff mechanisms can be valuable for the exploitation and improvement of physical based hydrological models. Moreover, the model can also improve the experimental and runoff mechanism researches. A review of above three aspects are summarized in this paper. PMID- 12722453 TI - [A review on mast seeding studies]. AB - Mast seeding means that the plants of a population have a highly variable seed production in different years synchronously, and it is widely existed in perennial plants. The periodic variation of seed yield and synchrony of seed production among individuals are the key characteristics. Several hypotheses, including evolutionary bases and limitations of mast seeding, were reviewed in this paper. Moreover, the synchronous degree among individuals was discussed, and the temporal permanence and spatial heterogeneity were emphasized. PMID- 12722454 TI - [Temporal-spatial scale characteristic of wetland ecosystem health]. AB - Wetland ecosystem health (WEH) is a new concept in wetland researches. Beginning with basic concepts and characters of temporal-spatial scale, all kinds of characteristics of WEH under temporal scale, spatial scale and temporal-spatial scale were discussed in this paper, and the significance of temporal-spatial scale in the researches of WEH was emphasized. PMID- 12722455 TI - [Characteristics and environmental significance of soil dissolved organic matter]. AB - Soil is a complex ecosystem with multi-interface. A numerous studies on soil dissolved organic matter (DOM) were carried out, and proved that DOM was one of the most active chemical components in the environment. Increasing attention has been paid on the study of soil DOM, especially in recent years, and the study has become an interdisciplinary focus in the fields of soil science, ecology, and environmental science due to the important roles of DOM in the biogeochemical cycles of carbon, nitrogen, phosphorus, sulfur, etc. In addition, DOM has significant effects on pedogenesis, growth and metabolism of soil microorganisms, decomposition and transformation of soil organic matter, and transport of pollutants in soils. The recent literatures about extraction methods, origin, composition, contents and controlling factors, bioavailability, and environmental significance of DOM were therefore reviewed, and future research aspects on this topic were also proposed. PMID- 12722456 TI - [Soil quality and its evaluation]. AB - Preservation and improvement of soil quality is the basic task for agricultural sustainable development. For this reason, the concept and the evaluation method of soil quality should be clearly understood. In this paper, the concept of soil quality and its advanced researches were reviewed. Soil quality evaluation should be based on soil function, different types of soils should be evaluated by different criterions, and soil quality evaluation should be relational rather than absolute as well. Some methods of soil quality evaluation were introduced, and the selection of evaluation indicators was discussed. PMID- 12722457 TI - [Discussion on freshwater ecosystem service and its evaluation index system]. AB - The importance of freshwater ecosystem and the connotation of ecosystem service were expounded in this paper, and the insufficiency in studies on freshwater ecosystem service was pointed out. Freshwater ecosystem service cannot be fulfilled without ecosystem health, yet there is a scarcity of synthetic studies integrating freshwater ecosystem service with ecosystem health, while these studies are necessary and important. Water resources should include water capacity, water quality, water energy, and aquatic organism. Based on the new meaning of water resources, the freshwater ecosystem service and its evaluation index system were discussed, and the evaluating methods were introduced briefly. The consideration and technical course of developing studies on freshwater ecosystem service in China were also discussed. PMID- 12722458 TI - [Research advance in estuarine phytoplankton ecology]. AB - The estuarine phytoplankton composition, spatial and temporal distribution, primary productivity, and their influencing factors were reviewed, and the significance of estuarine phytoplankton in monitoring aquatic environment, biodiversity of estuarine phytoplankton, and edge effect were discussed. Generally, the dominant phytoplankton populations were diatoms and dinoflagellates in estuaries. Microphytoplankton and picophytoplankton played important roles in estuarine ecosystems. Spatial and temporal changes in estuarine phytoplankton species composition and primary productivity were apparent. Several factors including light, temperature, nutrients, herbivory and river flow could influence estuarine phytoplankton production. PMID- 12722460 TI - [Micro-ecology in animal stomach and digestive tracts--theory and practice]. AB - The origin, development and practical use of animal micro-ecology community in the gastrointestinal tract of domestic animals were reviewed, and the microbial species, features, and their function mechanism were introduced. Meanwhile, the relationship of micro-ecological theory, microbial additives and their practical use in animal feeding were emphatically discussed. PMID- 12722459 TI - [Roles of rhizosphere in remediation of contaminated soils and its mechanisms]. AB - Rhizosphere is a special 'ecological remediation unit' to treat contaminated soils, which contains a great quantity of microorganisms such as fungi and rhizobacteria living with plant roots. Thus, physiological and ecological roles of plant roots to remedy contaminated soils, to accumulate and to solidify heavy metals, to absorb and degrade organic pollutants in rhizosphere were illustrated, and the function of mycorrhizospheric fungi to absorb, barrier and chelate heavy metals, to degrade organic pollutants through their metabolism activities, the action of rhizobacteria to absorb and solidify heavy metals, to degrade organic pollutants in rhizosphere through their active living activities, and the combined remediation of fungi and bacteria to pollutants in rhizosphere and their relevant mechanisms were explained. It was suggested that the remediation role of rhizosphere was the main part of phytoremediation, and one of the main basic theories to remedy contaminated soils by the activity of green plants and other organisms. The use of hyperaccumulative plants in remedying soils contaminated by some heavy metals would be prospective. It would be one of the important approach to contaminated soils remediation by organic pollutants through the mechanism of screening some special plants whose roots had strong secreting ability to absorb and accumulate dissolvent organic pollutants on the basis of inoculating specific or non-specific fungi and bacteria from the rhizosphere. This will be a developing trend of research on the remediation of contaminated soils by organic pollutants. PMID- 12722461 TI - [Analysis on leaf forms in Quercus mongolica community]. AB - Leaf-size classes of 337 vascular plants in Quercus mongolica community in northeast, China were analyzed according to Raunkiaer system. The dominant type was microphyll which percentage is 45.4%, followed by compound leaf (23.1%), mesophyll (22.6%), nanophyll (6.8%), macrophyll (0.9%), aphyllous (0.9%), leptophyll (0.3%). The leaves edges of 337 plants were investigated and the percentage of entire leaves plants is 22.3%. Leaf-size classes spectra of trees, shrubs and herbs were figured out. The mesophyll ratio in trees was higher than that in shrubs and herbs. The mesophyll ratio in shrubs was the lowest. Different sites had different leaf-size classes spectra. Microphyll also was the dominant type in every site. No significant relationships were found between leaf-size classes spectra and latitude and altitude. PMID- 12722462 TI - [Litterfalls of major forest stands at Baiyunshan scenic spot of Guangzhou]. AB - The productions, seasonal dynamics, macronutrient contents and decomposition rates of the litterfalls of four typical stands, e.g., Pinus massoniana plantation, secondary evergreen broadleaved forest, Acacia mangium plantation and Schima superba-Acacia mangium plantation at the scenic Baiyunshan of Guangzhou were studied. The litterfall productions of four stands in 1998 were 8.34, 6.77, 6.31 and 11.54 t.hm-2, respectively. The seasonal dynamics of litterfall amounts demonstrated the single-peak model with the peak period in June and July. The total amounts of macronutrients returned to the forest land by means of litters in the four stands in 1998 were 26.30, 69.81, 54.9 and 152.24 kg.hm-2, respectively. The annual decomposition rates of the litterfalls were 24.4%, 53.4%, 52.4% and 57.4%. PMID- 12722463 TI - [Soil nutrient, microorganism and enzyme activity in Pinus elliottii mixed stands]. AB - Soil nutrients, microorganisms and enzyme activities in mixed stands of Castanopsis fissa x Pinus elliottii and Schima wallichii x Pinus elliottii were studied. The results showed that compared with pure Pinus elliottii stand, Pinus elliottii mixed stands could increase organic matter content and improve the nutrient status of soil, especially for available nutrients of soil. The amounts of bacteria, actinomyces and enzyme activity in Pinus elliottii mixed stand were higher than those in pure Pinus elliottii stand, whereas their fungi amount was lower than the latter. PMID- 12722464 TI - [Arteries of the hilum and parenchymal part of the ovary in reproductive age in microangiographic studies]. AB - OBJECTIVES: The main aim of ours research was to recognize the arterial system of female ovaries in women in the reproductive age. MATERIALS AND METHODS: Study was performed on 31 cases of organs taken from the minor pelvis, composing female genital organs, urinary bladder, rectum and the upper part of the vagina from women aged 15 to 40. The ovarian and the uterine arteries have been injected with fine-granular radiological contrast medium Hexabrix 200 and Ultravist 370. Global X-rays of the whole organs complex and the microangiography of the isolated uterine adnexa and ovarian glands scraps were made. RESULTS: In all the examined cases the main source of ovarian blood supply was the ovarian arch created by the ending of the ovarian artery and the ovarian branch of the uterine artery. A different number of branches of the main vessel of the ovarian arch were going into the hilum of the ovary gland. The arterial branches on all length of the ovarian arch have been disposed as a dense net. Nearly all vessels of the ovarian arcade after short course have been divided several times and finally a higher number of branches were getting into the parenchymal parts of the ovarian gland. The vessels of the parenchymal part have been arranged regularly in the middle of the gland from the hilum to the free edge. In ends and peripheral parts of the ovary gland the number of vessels were significantly reduced. There were singles thinly vessels with non-typical course only. CONCLUSIONS: The arteries of the uterine adnexa always come from two sources: ovarian artery and uterine artery, both are forming the ovarian arch. Arteries of ovarian medulla are running axially, from hilus to the free margin of the gland. The spiral arteries system of the gonad is uniform and dense in the middle part of the gland, peripheral parts of the ovary and the area of ends of the gonads have got poor vessel picture. PMID- 12722465 TI - [Arteries of the uterine cervix in reproductive age in microangiographic studies]. AB - OBJECTIVES: The main aim of our research was the estimation of cervical arteries in women in the reproductive age. MATERIALS AND METHODS: Study was performed on 61 cases of organs taken, during autopsies, from the minor pelvis composing the female genital organs, urinary bladder, rectum and the upper part of the vagina from women aged 18 to 41. The material was divided in two groups: mothers and non mothers. The uterine arteries of a fresh specimen have been injected with radiological contrast medium Hexabrix 200 and Ultravist 370. The global X-rays of the organ complex and the microangiography of the uterine cervix were made. After that uterus was sectioned horizontally in sections of 4-5 mm thick and series of x-rays was performed. The number, size and way of uterine artery branches in the cervix were estimated. RESULTS: The cervical branches of the uterine artery were present in every cases. Usually branches were from 1 to 4 in number. All the branches were divided in cervical walls, very rich in upper and poorer in lower part of organ. Radiographic analysis revealed that connections between cervical vessels on both sides of the same uterus weren't permanent. Their absence was associated with the presence of the hypovascular or even avascular zone in the middle of the anterior and posterior cervical wall. CONCLUSION: The cervical branches of both uterine arteries are always present in the reproductive age, Number, size, course and way of cervical branches are different in all parts of the organ. The angioarchtectonic of cervical vessels does not depend on past pregnancies. PMID- 12722466 TI - [Results of cervical cancer treatment at the Lower Silesian Oncology Center between 1987 and 1989]. AB - OBJECTIVE: The aim of the project was to analyze a survival of patients with cervical cancer during 10-years follow-up and to identify factors that predict survival. Age, place of living (city vs. country), stage of disease, histology of the tumor and performed treatment were considered as potential prognostic factors. MATERIALS AND METHODS: The analysis included 497 patients diagnosed with cervical cancer, treated in Lower Silesian Oncology Centre between 1987 and 1989. Squamous cell carcinoma was a predominant type of the tumors. 488 patients were treated with radiotherapy, 92 patients underwent hysterectomy m. Freund. Intracavitary brachytherapy with radium 226 or cesium 137 (remote after loading- technique) was performed in most (486) patients. RESULTS: 10-years survival rate was 43.5% (216 patients), 5-years-survival 53.3%. During 5-year follow-up mortality decreased more rapidly than during next 5 years. It has been observed, that prognosis was better for younger patients (< 50 years old) with 10-y SR (survival rate) of 51.9%, compared with the group of older (> 50 years old) with 10-y SR of 35.5%. No difference in survival was noticed between patents living in the cities and in the country. Like other, I found that clinical stage of disease is the most important prognostic factor based on observed SR. 5-y SR was 72.5%, 59.5%, 22.1% an 10-y SR was 63.8%, 48%, 15.2% for stage Ib, IIb, IIIb respectively. Brachytherapy with cesium resulted in improved SR of 50.9% compared with radium (39.8%). CONCLUSION: Based on obtained data, the factors of prognostic value are: stage of disease, age and treatment. PMID- 12722467 TI - [Nucleolus organizer regions (agnors) and Ki-67 antigens expression from the cervix and cervical canal smears with respect to changes in ascus and AGUS (atypical cells of undetermined significance) according to Bethesda Cytological System of Classification]. AB - OBJECTIVES: The cytologic report written by a cytopathologist on the basis of smears from the cervix and cervical canal ought to give detailed and worthwhile information which allows making appropriate diagnostic and therapeutic decisions. The diagnosis of atypical cells of unspecified importance (ASCUS and AGUS) ought to have clinicians to carry out further diagnostic histopathologic procedures in order to explain the pathologic character of processes going on in the female reproductive organ. This is because such pathological states may as well spell could mean reactive, preinvasive as neoplastic changes of invasive character. DESIGN: This problem has led to carry out a research in order to examine the proliferative potential of cells in cytological smears (Ki-67, AgNORs) and to answer whether this potential allows us to qualify atypical cells as changes of reactive or neoplastic type. MATERIALS: In our study there were estimated 97 cases of cytological smears from the cervix and cervical canal. In this paper, immunohistochemical methods, and staining techniques developed by Ploton et al. in the detection of AgNORs and in the examination of Ki-67 antigen were used. CONCLUSIONS: 1. The average number of AgNORs in cell nucleus was highest in ASCUS cells and exceedingly higher in CIN 1 and CIN 3.2. The areal ratio of AgNORs to that of the cell nucleus was highest in HSIL and cancer cells, least in ASCUS. 3. In ASCUS and LSIL, AgNORs were minute and scattered whereas they were large and centrally located in HSIL and cancer cells. RESULTS: We found that analysis of AgNORs makes easier the differentiation of ASCUS and LSIL cells from HSIL and neoplastic ones according to the classification system of Bethesda. PMID- 12722468 TI - [Modulation of hypothalamo-pituitary axis by stress during labor]. AB - OBJECTIVES: The present study is aimed to investigate the function of hypothalamo pituitary-adrenal axis of women during late pregnancy and term labor. DESIGN: Levels of hormones were measured in blood of 34 women undergoing spontaneous labor and elective cesarean section, 2 days before and after delivery, and during labour. Additionally, blood from the umbilical vein and artery was also collected. MATERIALS AND METHODS: We have evaluated changes in corticotropin releasing hormone (CRH), adrenocorticotropin (ACTH) and dehydroepiandrosterone (DHEA) in vein blood of 34 subjects. The concentrations of hormones were measured by dint of RIA method. RESULTS: No significant correlation was found between hormone measurements and fetal outcome. CRH level in the umbilical vein was higher than in the umbilical artery, suggesting the placental origin of hormone. Prepartum CRH concentration was significantly higher in the group of spontaneously delivered patients. There were no correlations between CRH levels and ACTH and DHEA concentration in mother's blood plasma. In fetuses, higher prepartum CRH concentrations resulted in elevated levels of ACTH. No changes were found in DHEA concentration, in both mother and fetus. CONCLUSIONS: These results suggest that placental CRH may modulate a fetus's pituitary but not mother's. The observed high levels of this hormone play an important role mainly in preparation of mother and fetus for delivery. PMID- 12722469 TI - [New methods of monitoring the fetus during labor--detection of intrapartum fetal hypoxia by electrocardiography]. AB - The biophysical diagnostic methods are an essential element of evaluation of the fetus state in pregnancy and delivery. Nowadays, the applied methods of fetus supervision during labour don't ensure the permanent analysis and they don't deliver diagnostic information about fetus hypoxia. With a view to avoiding the risk of fetus injury and its hypoxia technologies should be developed which allow to make a diagnosis of perils to good fetus condition and detection of intralabor hypoxia. Therefore, enhancing all new methods are indicated, which facilitate the diagnostic in a permanent mode and which allow for hypoxia prediction and the reduction of unjustified medical interferences. One of these methods is fetus electrocardiography. PMID- 12722470 TI - [Curettage immediately after delivery]. AB - The purpose of the study was to compare indications for curettage directly after delivery with histologic findings. Only in 51% there was a histologic confirmation of placental or membrane tissue. Residences indications of the suspected retained placenta were wrong in 70% cases. The curetting of 75% of the patients experiencing post partum bleeding revealed no fragments of placental or membrane tissue. PMID- 12722471 TI - [Two cases of congenital toxoplasmosis with a central nervous system damage- delayed diagnosis]. AB - OBJECTIVES: The intrauterine toxoplasma gondii infection is linked with a high risk of central nervous system/CNS/damage in the fetus. Why, despite the wide knowledge of this neurodegenerative disease prophylaxis and therapy, do we still meet neonates with the CNS damage? DESIGN, MATERIALS AND METHODS: Authors present two cases of congenital toxoplasmosis with the CNS involvement. RESULTS: One of the cases was identified as the internal hydrocephalus during a prenatal ultrasound examination but no further diagnostics tests were undertaken. The congenital toxoplasmosis and severe CNS damage was diagnosed post delivery. In the other case some clinical symptoms of the CNS infection appeared in the neonatal period. Now this patient is eight months old and presenting mild motor developmental delays. CONCLUSIONS: 1. There are many clinical symptoms of the congenital toxoplasmosis, that can occur in the prenatal/case 1/and postnatal/case 2/period. 2. No serologic tests for toxoplasmosis were performed in mothers during pregnancy, what delays treatment and diagnosis of sick children. 3. Fetus presenting the CNS anomalies identified during ultrasound examinations should be immediately diagnosed. PMID- 12722472 TI - Gliomatosis peritonei with mature teratoma of the ovary. AB - We report a case of menstrual mature teratoma of ovary with implants of glial tissue in peritoneum and its adnexa. After resection of the tumour and omentum laparoscopic examination was performed and revealed reduction, fibrosis of glial implants and massive cellular reaction. PMID- 12722473 TI - [Delayed delivery of the second twin--a case report]. AB - Delayed interval delivery of the second twin is not a conventional procedure in case of early delivery of the first one. The routine procedure is the provocation of the uterine activity and terminating pregnancy. Patient's willingness to have a desired child presents difficulties for obstetricians and necessitates unconventional steps. The described delivery of the second twin took place in the fifth week after the delivery of the first one. The patient was discharged on the 7th day after caesarean section and the new-born was discharged in good general condition on the 64th day when reaching body weight 3490 gram. Postponing the delivery of the second twin gave time for treatment with glucocorticoids and it prepared the fetus's lungs for extrauterine life. Therapeutically the most essential problem was the prevention of intrauterine infection. The described procedure posing a threat to both mother and foetus gives rise to understandable doubts. Possible risks should be thoroughly discussed with the patient as well as possible future child's problems. PMID- 12722475 TI - [Genital mucous membrane involvement in the course of autoimmune bullous diseases -a diagnostic problem]. AB - Blisters and erosions localized to genital mucous membranes often accompany various diseases and may cause significant diagnostic difficulties. Autoimmune bullous diseases (pemphigus, pemphigoid, LABD, EBA), characterized by skin and mucosal lesions, sometimes are limited only to genital mucous membranes. The possibility of the autoimmune bullous disorders should be considered in the differential diagnosis of genital tract ulceration because of severity of those diseases and the necessity of long-term immunosuppressive therapy. The correct diagnosis of autoimmune bullous diseases is established by immunofluorescence tests (DIF, IIF). PMID- 12722474 TI - [Melatonin and its role in human reproduction]. AB - Melatonin may be a key factor in regulation of seasonal variation in gonadal activity. The circadian disturbances related to reproduction are probably subsequent to the seasonal change. Moreover, melatonin might also be considered essential to both spermatogenesis and folliculogenesis. Exposure to bright light, suppressing the concentration of melatonin in circulation, is hypothesized to be useful in treatment of both male and female infertility in couples with abnormal melatonin metabolism. In women, seasonal variation observed in the concentrations of gonadotropins and gonadal steroids is associated with melatonin synthesis. This hormone may exert an inhibitory effect on the GnRH pulse generator to decrease gonadotropin secretion. Consequently, it has been implicated in the control of the pubertal onset and timing of ovulation. The number of irregular menstrual cycles with anovulation in creases in winter compared to summer. In addition, there is circadian variation in the time of ovulation, occurring usually in the morning during summer and in the evening during winter. Hormonal interactions and phases of the cycle are important variables contributing to the fluctuations in melatonin levels during the menstrual cycle. PMID- 12722476 TI - The karyotypic structure of cell populations in vitro as an integral system. AB - This review describes regularities of karyotypic variability maintaining karyotypic stabilization of continuous cell lines. Statistical analysis of individual karyotypes of "marker" and "markerless" cell lines show that survival of cell population in vitro is maintained by a certain ratio of cells with different structural variants of karyotype (SVK). Characteristic feature of karyotypic variability in the "markerless" cell lines during long-term cultivation under various conditions is dicentric formation due to telomeric associations. These dicentrics seem to form genetical structures providing adaptation to conditions in vitro of the cell population as an autonomous system. Correlations between the numerical variability reflecting in SVK, and structural variability (dicentric formation) are manifestations of an integral cell populational function. Experimental data allow to suggest that integrity of the karyotypic structure of cell populations is maintained not only by selection of random variations, but also by programmed (adaptive) changes of karyotype. As a whole, in the cell population the state is realized that can be called karyotypic homeostasis; the observed phenomena characterize processes maintaining such homeostasis. PMID- 12722477 TI - [Locomotion of CV-1 cytoplasts with or without a centrosome]. AB - The movement of cultured cells along the substratum is a convenient model for studying cell movement in the organism, occurring during embryogenesis, angiogenesis, metastasis, wound closure, etc. The moving cells must control their pseudopodial activity along the perimeter, regulate the attachment and reattachment to the substratum, and pull their body following pseudopodium during their movement along the substratum. As proven by numerous investigations, these processes directly depend on the actomyosin system of cells. The role of microtubules as components of cytoskeleton in cell locomotion still remains obscure. The role of microtubules in cell movement is commonly investigated using microtubule-destructive drugs. Therefore in the final results the accessory drug effect on, for example, signal cascades cannot be excluded. Another mode of action on the microtubule dynamics is centrosome removal from the cells, which is easily realized by their removal together with the nucleus. It has been shown that in cytoplasts of centrosome containing fibroblasts, dynamic instability of microtubules remains. Unlike, in non-centriolar cytoplasts tread milling is observed. Some literature evidence suggests that cytoplasts of cultured cells move along the substratum not worse that intact cells do. In this study cytoplasts with and without centrosome were obtained and identified, and parameters of movement along the substratum (speed, direction) were registered for both these two populations of cytoplasts, and for control intact cells and cells involved in the experiment. The model of experimental wound of monolayer was used, because it guaranteed cell synchronization in respect to movement direction and speed. Centrosome-containing CV-1 cytoplasts displayed radial microtubule array, and centrosome-lacking cytoplasts exhibited chaotic distribution of microtubules, which is characteristic of microtubule tread milling. In addition, both kinds of cytoplasts appeared to move along the substratum much slower than the whole cells. No difference was found is speed and keeping direction between centriolar and non-centriolar cytoplasts. PMID- 12722478 TI - [Effects of actin cytoskeleton rearrangements on channel activation by calcium entry into A431 cells]. AB - Using patch clamp and ion-selective fluorescence dye techniques, we investigated the influence of actin cytoskeleton rearrangements on the activity of calcium entry channels in plasma membrane of human carcinoma A431 cells. It is shown that disruption of actin microfilaments by cytohalasin D has no significant effect on calcium release from the stores and its entry from the extracellular space. It also does not interfere with the activation of inositol 1,4,5-trisphosphate (IP3) dependent high-selective low-conductance calcium channels Imin. The treatment of cells with calyculin A induces the formation of actin filament layer beneath plasma membrane and also inhibits Imin activation and calcium entry through the plasma membrane, though calcium efflux from the stores was nearly unchanged. Thus, it is concluded that calcium signalling in A431 cells can be modulated by actin cytoskeleton rearrangements, and may be well described in terms of "conformational coupling" model. PMID- 12722479 TI - [Changes in the activity of cyclin-kinase complexes governing cell transition from G1 phase to DNA replication phase in E1A + c-Ha-ras transformants transfected with the bcl-2 gene]. AB - The antiproliferative effect of human bcl-2 gene transferred to E1A + c-Ha-ras transformed rat embryo fibroblasts, which are characterized by the absence of cell cycle checkpoints after damage and by a high proapoptotic sensitivity was studied. Ionizing irradiation, adriamycin treatment, and serum starvation were shown to induce G1/S arrest in E1A + c-Ha-ras-transformants. Bcl-2 antiproliferative effect in E1A + c-Ha-ras-transformants was not associated with alterations in Cdk2, cyclin E and A contents. G1/S arrest following irradiation or serum starvation was accompanied by a decrease in kinase activity associated with cyclin E-cdk2, whereas G1/S arrest in tetraploid subpopulation after adriamycin treatment did not correlate with a decrease in cyclin E-associated kinase activity. Cyclin A-associated kinase activity did not decrease after any used treatment. Transfection of bcl-2 in E1A + c-Ha-ras-transformants resulted in elevated expression of cyclin-cdk complexes inhibitor p21/Waf-1, but not p27/Kip. Damaging agents caused p21/Waf-1 and p27/Kip accumulation, but bcl-2 overexpression did not restore functions of these inhibitors, since p21/Waf-1 and p27/Kip were unable to suppress cyclin-cdk complexes activity after damage. These results suggest that bcl-2 transfection in E1A + c-Ha-ras-transformants is likely to result in irradiation- or serum starvation-induced G1/S arrest accomplished by a selective decrease in cyclin E-associated kinase activity. Adriamycin-induced G1/S arrest seems to be realized via cyclin-cdk complexes activity-independent way involving antiproliferative targets downstream of cyclin E-cdk2 and cyclin A cdk2 complexes. PMID- 12722480 TI - [EGF-induced signal transduction in clonal cells of epidermoid carcinoma A431]. AB - A431 epidermoid carcinoma cells have an increased expression of EGF receptor. In contrast to many other cell lines and primary cells, these respond to EGF in high (more than 1 ng/ml) concentrations by cell cycle arrest, apoptosis and detachment. Clonal variants of A431 (1a and 8a), able to grow in the presence of EGF in high concentration, were previously developed in our laboratory (Gudkova, Sorokin, 1989). Here we tested upper pathways of signal transduction from EGF receptor in the clonal variants, as compared to A431. We found no reasonable difference in the expression of EGF receptor, as well as in its EGF-induced phosphorylation in A431 and clonal variants. There were also no changes in the amount and activation of ERK MAP kinase in different cell lines. In contrast, the amount of STAT1 transcription factor, known to play pro-apoptotic and antiproliferative roles, was strictly diminished in both the clonal variants tested (1a and 8a), as compared to the parental line A431. However, EGF-induced tyrosine phosphorylation of STAT1 decreased only in 8a. Increased phosphorylation of Akt protein kinase, the key component of PI-3 kinase of the anti-apoptotic and proliferative signaling pathway, was also observed in clonal variants. The data obtained demonstrate that resistance to EGF can be acquired in cells having similar levels of EGF receptor expression and phosphorylation, but different in STAT1 or PI-3 kinase signal transduction pathways. These pathways may presumably represent two antagonistic key elements regulating A431 proliferation and apoptosis. PMID- 12722481 TI - [RNA polymerase II and pre-mRNA splicing factors in diplotene oocyte nuclei of the giant African gastropod Achatina fulica]. AB - The nuclear distribution of pre-mRNA splicing factors (snRNPs and SR-protein SC35) and unphosphorylated from of RNA polymerase II (Pol II) was studied using fluorescent and immunoelectron cytochemistry in diplotene oocytes of the gastropod Achatina fulica. Association of Pol II and splicing factors with oocyte nuclear structures was analysed. The antibodies against splicing factors and Pol II were shown to label perichromatin fibrils at the periphery of condensed chromatin blocks as well as those in interchromatin regions of nucleoplasm. The revealed character of distribution of snRNPs, SC35 protein, and Pol II, together with the decondensed chromatin and absence of karyosphere, enable us to suggest that oocyte chromosomes maintain their transcriptional activity at the diplotene stage of oogenesis. In A. fulica oocytes, sparse nuclear bodies (NBs) of a complex morphological structure were revealed. These NBs contain snRNPs rather than SC35 protein. NBs are associated with a fibrogranular material (FGM), which contains SC35 protein. No snRNPs were revealed in this material. Homology of A. fulica oocyte nuclear structures to Cajal bodies and interchromatin granule clusters is discussed. PMID- 12722482 TI - [Changes in the expression of membrane markers and in the number of human blood monocytes after single and repeated courses of visible and infrared light at therapeutic doses]. AB - An attempt has been made to prove that the immunomodulating effect of therapeutic doses of polychromatic visible + infrared polarized (VIP) light at its application to a small body surface area is connected with a transcutaneous photomodification of a small amount of blood in superficial skin microvessels. For this purpose, in parallel experiments, using monoclonal antibodies, the membrane phenotype of circulating blood mononuclears was studied after irradiation of volunteers, of samples of their blood in vitvo, and of a mixture of the irradiated and non-irradiated autologous blood in a 1:10 volume ratio, thereby modeling events in vivo, when a small amount of the transcutaneously photomodified blood in the vascular bed contacts its main circulating volume. In this variant of experiment, a great similarity has been established of changes in expression of mononuclear membrane markers (CD3, CD4, CD8, CD20, CD16, HLA-DR and to a lesser degree of CD25); the ability has been proven of the photomodified blood to "translate" the light-induced changes to a much higher volume of non irradiated blood, which might represent a mechanism of the systemic immunomodulating effect of phototherapy. Under conditions in vivo and in vitro, the most "reactive" were HLA-DR+, CD20+, CD16+, CD4+, and 0-cells. An increase of the total number of lymphocytes and monocytes has been shown by the end of the 10 day-long phototherapeutic course. The regulatory character of the single and course sessions of the VIP light on the blood immunocompetent cells is substantiated: depending on the initial state of the immune system, the VIP light can produce both stimulating and inhibitory effect on lymphoid cell subpopulations, which opens large possibilities of using this method for correction of immunological disturbances in diseases of different etiopathogenesis. PMID- 12722483 TI - [The role of tyrosine phosphorylation in the regulation of cell proliferation and differentiation in lower eukaryotes]. AB - The review summarizes current data on signaling transduction mechanisms in some unicellular eukaryotes, based on sequential translation of phosphotyrosine signals from the cell surface to the nucleus. PMID- 12722484 TI - Pastoral crisis intervention in response to terrorism. AB - Pastoral crisis intervention may be thought of as the functional integration of crisis intervention and pastoral support. In effect, the practice of pastoral crisis intervention largely represents the use of faith-based interventions refined and augmented through the use of an emergency mental health delivery context. The value of pastoral crisis intervention seems apparent in situations involving death, serious injury, mass disasters, and cataclysmic events such as war. Nowhere, however, is pastoral crisis intervention potentially more useful than in response to real or threatened terrorism. PMID- 12722485 TI - A maritime disaster: reactions and follow-up. AB - In 1999, 69 people survived a maritime disaster on the Norwegian coast, during which 16 others died. Besides immediate psychosocial assistance, post-disaster intervention included psychological debriefings after one week, follow-up debriefing a month later, screening of those in need of individual help, and help for those returning to the scene of the disaster. The results of the psychometric tests showed that a considerable number of survivors scored above clinical cut off points for extreme stress reactions. These results were compared with results from other studies of maritime disasters. Although the life threat and exposure in this disaster were extreme, the scores were lower than for the other studies, with one exception. The authors concluded the lower distress scores compared to other maritime disasters were probably impacted by the structured and caring system that was implemented to care for survivors. Almost all (93%) considered the debriefing meetings as helpful, and they were able to discriminate between different functions served by the meetings. PMID- 12722486 TI - PSTD, major depressive symptoms, and substance abuse following September 11, 2001, in a midwestern university population. AB - This research investigated the prevalence of Posttraumatic Stress Disorder (PTSD), Major Depressive Disorder (MDD) and substance abuse in a midwestern university population following the terrorist attacks on September 11, 2001, in New York City and Washington, DC. Three-hundred five subjects volunteered to complete a questionnaire which measured nine areas of psychosocial functioning which included demographics, personality, PTSD, MDD, prior traumatic experiences, alcohol and drug use, psychiatric history and treatment, and current attitudes towards government, religion, the economy, and how children were affected by the events. The participants lived in a large urban city over which United Flight 93 circled before crashing in Pennsylvania due to terrorist attacks. The subjects were forced to evacuate their university and city due to attacks on New York and errant United Flight 93. The study also replicated the first two national studies on PTSD prevalence (Schuster, et al., 2002; Galea, et al., 2002). The results found a prevalence rate of 5.9% for probable PTSD, matching identically previous national surveys. There were higher levels of PTSD and MDD for females, those with less education and who were single or unmarried, and those who had a prior history of mental health problems or psychological trauma. PTSD and MDD were associated with higher levels of alcohol and drug use since September 11. Relations to active duty military personnel appear to moderate the perception of threat, suggesting the importance of affiliative kinship patterns to coping with stress. Finally, the concept of geographic and psychological proximity to the 'zone of danger' is discussed. PMID- 12722488 TI - Compassion satisfaction, compassion fatigue, and critical incident stress management. AB - This study examines the potential for compassion satisfaction, compassion fatigue, and burnout in a sample of persons attending a workshop on Prevention of Compassion Fatigue at an international conference of providers of Critical Incident Stress Management (CISM) services. In the study, more than half (58%) of the respondents reported experiencing psychological reactions after providing CISM services, including an array of behavioral, emotional, cognitive, and physical symptoms of psychological stress. Forty percent of respondents were found to have moderate, high, or extremely high risk for compassion fatigue. At the same time, 89% of respondents were found to have a good, high, or extremely high potential for compassion satisfaction, and 87% were found to be at extremely low risk for burnout. The results appear to indicate that, while the CISM practitioners recognize the stress associated with their work (as reflected in the reported symptoms), the work provides significant rewards (as measured by compassion satisfaction) that outweigh the stress and mitigate against burnout. Likewise, while 40% tested positive for compassion fatigue (or secondary traumatic stress) as a result of their empathy with CISM recipients, the rewards of the work again appear to mitigate the negative effects of the work. Implications for future research and practice are presented. PMID- 12722487 TI - An evaluation of the Respiratory One Method (ROM) in reducing emotional exhaustion among family physician residents. AB - The purpose of this study was to evaluate a particular burnout risk reduction method among family physicians within a family practice residency program and to contribute to the very limited research literature on family physician residents' stress reduction. The author used the Respiratory One Method (ROM) as a burnout risk reduction method. This study used a quasi-experimental design to compare participating physicians with a control group. To test the research hypothesis that ROM has an effect on emotional exhaustion (EE), the data were analyzed using the Mann-Whitney U test. The results revealed that the ROM treatment did have an effect on emotional exhaustion scores. Findings of this study are useful for burnout prevention and curriculum development among family physicians caring for patients in family practice programs. PMID- 12722489 TI - Coping with threats of terrorism: a protocol for group intervention. AB - This article presents a group protocol designed to assist people in coping with direct and ongoing threats of terrorism. The protocol is intended to enable participants to address the psychological issues necessary to cope during periods of extreme threat. A step-by-step description of the protocol is provided. PMID- 12722490 TI - The Emperor has no clothes: velocity, flow, and the use of TCD. PMID- 12722491 TI - B0 images obtained from diffusion-weighted echo planar sequences for the detection of intracerebral bleeds. AB - BACKGROUND AND PURPOSE: To evaluate the accuracy of B0 echo planar imaging (EPI) sequences for the detection of intracerebral bleeds. METHODS: One hundred patients with acute strokes had magnetic resonance imaging and computed tomography (CT) examinations performed within 48 hours after the onset of symptoms. The detectability of intracerebral bleeds by the B0 EPI sequences was assessed. The results were compared to the gradient echo (GRE) sequence and CT brain examinations. The results of the GRE sequences were used as the gold standard. RESULTS: The B0 EPI sequences detected 11 out of 11 acute, intracerebral hematomas; 6 out of 8 acute hemorrhagic strokes; 2 out of 2 acute, intraventricular hemorrhages; 8 out of 8 old hemorrhagic infarcts; 1 out of 1 subarachnoid hemorrhages; and 11 out of 22 patients with microbleeds. For the detection of acute, intracerebral hematomas and acute, hemorrhagic infarcts, B0 EPI sequences had a sensitivity of 89.5%, a specificity of 100%, and an accuracy of 98%. CT had a sensitivity of 57.9%, a specificity of 100%, and an accuracy of 92%. B0 EPI sequences did not miss any acute or chronic hemorrhages detected by CT examinations. CONCLUSIONS: B0 EPI sequences could not replace GRE images for the detection of both acute and chronic hemorrhages. Their sensitivity for the detection of acute and chronic blood products, however, was comparable, if not superior, to that of CT examinations. PMID- 12722493 TI - Apolipoprotein E genotype and early Alzheimer's disease: a longitudinal SPECT study. AB - BACKGROUND AND PURPOSE: To determine the association of the apolipoprotein E (APOE) genotype and longitudinal changes of regional cerebral blood flow (rCBF) in Alzheimer's disease (AD). Previous reports have yielded conflicting results concerning this association. METHODS: A retrospective cohort study was performed. rCBF was noninvasively measured using 99mTc-ethyl cysteinate dimer single-photon emission computed tomography in 23 patients with probable AD at the very early stage and at a mean interval of 24 months, as well as in 55 age-matched healthy volunteers. Patients were classified into 2 groups according to the presence of the epsilon 4 allele: 11 epsilon 4 carriers and 12 noncarriers. Correction for partial volume effects (PVEs) was performed in all patients using gray matter volume measured by magnetic resonance imaging. Statistical parametric mapping was used for the analysis of absolute rCBF data and the adjusted rCBF images of relative flow distribution. RESULTS: In the baseline study, both carriers and noncarriers showed significant decreases of absolute and adjusted rCBF in the psoterior cingulate gyri and precunei. After PVE correction, carriers showed a greater spread of areas with significant rCBF reduction from the parietotemporal to the frontal area than noncarriers during the follow-up period compared to healthy volunteers. Moreover, carriers showed a significant decline of absolute rCBF in the frontal cortex from the baseline to the follow-up study. CONCLUSIONS: The authors' study suggests that the APOE epsilon 4 allele is associated with the faster progression of AD, and PVE correction may be necessary for accurate assessments of SPECT studies of AD. PMID- 12722492 TI - Cerebrovascular reactivity in hypertensive and healthy adolescents: TCD with vasodilatory challenge. AB - BACKGROUND: Adolescent hypertension predicts hypertension and increased cardiovascular morbidity in adulthood. The aim of the present work was to test whether cerebrovascular reactivity to CO2 is altered among hypertensive adolescents. METHODS: From the population-based cohort of the Debrecen Hypertension Study, 113 hypertensive and 58 normotensive adolescents underwent transcranial Doppler measurements at rest and after 30 seconds of breath holding. RESULTS: Systolic, mean, and diastolic blood flow velocities were higher among hypertensive individuals at rest. Taking the absolute blood flow velocity parameters into account, after breath holding, only the pulsatility index was significantly higher in the hypertensive group. The percentage change of the different blood flow parameters showed less intensive change in hypertensive teenagers. CONCLUSION: Cerebral vasoreactivity is decreased among hypertensive individuals as compared to healthy controls. PMID- 12722494 TI - Characteristics of microembolic signals detected near their origins in middle cerebral artery stenoses. AB - BACKGROUND AND PURPOSE: Usually, microembolic signal (MES) monitoring is performed for MESs arising from downstream sources. The aim of this study is to describe the special characteristics of MESs originating from nearby sources in the middle cerebral artery (MCA). METHODS: We analyzed 265 MESs detected in 5 patients (the study group) who had acute ischemic strokes within the territory of MCA stenoses and 48 MESs detected in 7 patients (the control group) from proximal sources: severe internal carotid artery stenoses and prosthetic heart valves. MESs were recorded with a 2-MHz, bigate transducer. We performed offline analyses of the recorded MESs using both post-fast Fourier transform (FFT) spectra and pre FFT time domain signals. MESs were divided into 3 types--focused-frequency signals (FFSs), bottom-frequency signals (BFSs), and multifrequency signals (MFSs)--according to the frequencies of the signals displayed on the post-FFT spectra. RESULTS: In the study group, among 265 MESs, 245 (92.5%) were MFSs, 11 (4.2%) were FFSs, and 9 (3.4%) were BFSs. In the control group, among 48 MESs, 45 (93.5%) were FFSs, 2 (4.2%) were MFSs, and 1 (2.%) was a BFS. There was significant difference in MES type between MCA stenoses and other embolic sources (P < .05). On the post-FFT spectra, MFSs recorded from MCA stenoses occupied multiple frequencies along the vertical axis, and the low-frequency parts were usually bidirectional. On the pre-FFT time domain signals, the highest frequency parts presented distorted, amplitude-modulated sine waves, and the low-frequency parts presented totally irregular waves. The amplitudes of the low-frequency parts diminished with time in the proximal channel but enlarged with time in the distal channel over a short period. CONCLUSION: MESs recorded from MCA stenoses may have special characteristics of multiple frequencies on both post-FFT spectra and pre-FFT time domain signals. Our findings may represent rotating or vibrating emboli as they are just dislodged from the thrombus and are moving from the vessel wall to the center. Further clinical and laboratory studies are needed to confirm this hypothesis. PMID- 12722495 TI - Measurement of carotid stenosis: comparisons between duplex and different angiographic grading methods. AB - BACKGROUND AND PURPOSE: Endarterectomy has been proved to be an effective stroke prevention procedure. However, there are still inconsistencies between the results of different preoperative evaluation methods, which may sometimes complicate treatment plans. This study measured the discrepancies between different angiographic grading methods and attempted to further assess the accuracy of the carotid duplex examination according to these different angiographic grading methods. METHODS: One hundred seventy-one preendarterectomy carotid duplex examinations and angiograms were reviewed. All angiograms were measured blindly by one of the authors using the North American Symptomatic Carotid Endarterectomy Trial (N), the European Carotid Surgery Trial (E), and the common carotid (C) methods. The measurement results were further converted into the area of stenosis indices (N2, E2, and C2, respectively). By using regression testing, all results could be compared. The duplex examination data were then compared with the results of different angiographic measurement methods to evaluate their accuracy. RESULTS: The measurement results of all angiographic grading methods were well correlated. Using different angiographic grading systems as the gold standard, duplex examination for screening endarterectomy candidates produced the following results: According to the N method, accuracy was 74%; according to the E method, accuracy was 90%; and according to the C method, the accuracy rate was 92%. According to N2, accuracy was 88%; according to E2, accuracy was 94%; and according to C2, the accuracy rate was 93%. CONCLUSIONS: The measurement results of all 3 commonly used angiographic grading methods were linearly correlated with one another. (It is important to note that none of those standards should be treated as the only gold standard.) In this study, the duplex criteria have a greater accuracy rate according to E or C rather than N. However, this study also demonstrated that the cut point of the Doppler criteria is the determinant factor for accuracy rather than which gold standard was compared. Through careful adjustment of the cutoff criteria, carotid duplex can be highly accurate, despite the use of different reference standards. PMID- 12722496 TI - Brain microembolism. AB - PURPOSE: To alert clinicians about the occurrence of a subtype of brain infarction, its suspected etiology, and its detection by specific neuroimaging techniques. METHODS: The article presents 5 nonconsecutive patients admitted to the stroke services of 2 tertiary care hospitals, who presented with acute or subacute symptoms suspicious, but at times atypical, of brain ischemia. FINDINGS: Each patient had evidence of 3 to > 20 small areas of recent brain infarction detected by diffusion-weighted imaging (DWI). When available, brain computerized tomography images were not helpful for the diagnosis of these recent infarcts. Most lesions were present on magnetic resonance imaging fluid-attenuated inversion recovery sequences, but the diffusion-weighted images allowed the determination of their acuity. Further evaluation revealed a potential source of embolism in each patient. Brain microembolism was suspected in all cases. CONCLUSION: Small and multiple areas of acute or subacute brain infarction occasionally present with clinical features atypical for brain embolism. They can be detected by magnetic resonance DWI studies. PMID- 12722497 TI - Diffusion-weighted MRI in Creutzfeldt-Jakob disease: a better diagnostic marker than CSF protein 14-3-3? AB - Two middle-aged patients presented with rapidly progressive dementia and ataxia, nonspecific electroencephalography findings, and negative cerebrospinal fluid (CSF) protein 14-3-3. Both patients underwent brain magnetic resonance imaging (MRI) scans that demonstrated abnormalities on diffusion-weighted imaging (DWI) sequences, and both were later confirmed to have Creutzfeldt-Jakob disease. (CJD) by tissue examination. Because a recent position paper from the American Academy of Neurology characterized CSF protein 14-3-3 as a gold standard for clinically diagnosing CJD, the authors reviewed studies of CJD in which DWI-MRI imaging and CSF protein 14-3-3 studies were both performed. Among 19 reported cases of CJD with DWI-MRI lesions, CSF protein 14-3-3 was negative in 6 cases and positive in 2 others. The authors' findings suggest that multifocal cortical and subcortical hyperintensities confined to gray matter regions in DWI-MRI may be a more useful noninvasive diagnostic marker for CJD than CSF protein 14-3-3. These observations provide a compelling rationale for a prospective comparative study. PMID- 12722498 TI - Reversal of perfusion and diffusion abnormalities after intravenous thrombolysis for a lacunar infarction. AB - Perfusion and diffusion magnetic resonance imaging (MRI) have been used to guide therapy in patients with large vessel occlusions. In an analogous fashion, the authors used perfusion and diffusion MRI to demonstrate a perfusion deficit and a matching diffusion lesion in a patient with lacunar infarct. Following thrombolytic therapy, the authors observed a reversal of the perfusion deficit, the diffusion lesion, and clinical recovery. This case suggests that perfusion and diffusion MRI may be informative in patients with lacunar infarction who are candidates for thrombolysis. PMID- 12722499 TI - Dynamics of dotlike hemosiderin spots associated with intracerebral hemorrhage. AB - The authors observed dotlike, low-intensity spots in T2*-weighted magnetic resonance imaging (MRI), subsequently diagnosed histologically as previous microbleeds associated with lipofibrohyalinosis, amyloid angiopathy, and small vessel disease. The nature of dotlike hemosiderin spots (dotHSs), however, is still unknown. This case report seeks to demonstrate the dynamics of dotHSs associated with an intracerebral hematoma (ICH). T2*-weighted MRI of a 72-year old man with a history of hypertension demonstrated 4 dotHSs 24 months after a left putaminal hemorrhage. Follow-up T2*-weighted MRI 40 months after the acute event demonstrated the asymptomatic formation of 3 more dotHSs, even with good control of blood pressure. Fifty months after the stroke, T2*-weighted MRI showed that 2 of the new dotHSs had become fainter, whereas the hemosiderin associated with the ICH scar remained detectable. To the authors' knowledge, this is the first description of dotHS dynamics associated with ICH. PMID- 12722500 TI - Acute stroke syndrome with fixed neurological deficit and false-negative diffusion-weighted imaging. AB - Diffusion-weighted imaging (DWI) is sensitive for the detection of acute ischemic stroke. However, a negative DWI study of the brain does not always exclude a patient from the possibility of acute cerebral ischemia. The authors report 1 case in which the patient presented with a fixed ischemic neurological deficit (National Institute of Health Stroke Scale score = 22) that included global aphasia, right hemiparesis, and a right visual field neglect. The initial DWI of the brain within 27 hours of symptom onset was negative. The deficit persisted, and a repeat magnetic resonance imaging study 7 days later showed a large area of restricted diffusion involving the gray matter of the entire left middle cerebral artery and anterior cerebral artery distribution, indicating a large area of cortical stroke. PMID- 12722501 TI - Water apparent diffusion coefficient and T2 changes in the acute stage of maple syrup urine disease: evidence of intramyelinic and vasogenic-interstitial edema. AB - BACKGROUND: The acute phase of the neonatal classical form of maple syrup urine disease (MSUD) is usually associated with generalized brain edema. METHODS AND RESULTS: The authors present the case of a newborn infant in the acute stage of the classical form of MSUD in whom a remarkable decrease in the water apparent diffusion coefficient (ADC) in advanced myelinating white matter areas was associated with an increase in the T2 signal. This diffusion magnetic resonance imaging (MRI) pattern appears to be compatible with a rare kind of cytotoxic edema, the so-called intramyelinic edema. At the same time, an increase in the ADC was seen in unmyelinated areas together with an increase in the T2 signal, a sign of a coexistent vasogenic-interstitial edema. CONCLUSIONS: ADC measurements in MSUD provide more specific information than conventional MRI about the pathophysiology of white matter changes. PMID- 12722503 TI - Nonpulsatile cerebral perfusion in Takayasu's arteritis. AB - A nonpulsatile cerebral perfusion may be encountered in an artificial cardiopulmonary bypass used in cardiac surgery, which is exceptional in physiological conditions. The authors report on a 37-year-old woman with Takayasu's arteritis (TA) who had been suffering from progressive visual loss and recurrent seizures. Ocular findings of chronic ischemia and multiple, subcortical, high-signal lesions in magnetic resonance (MR) imaging were indicative of significant hemodynamic impairment. MR angiography showed the complete occlusion of the innominate artery, the left common carotid artery (CCA), and the subclavian artery from the orifice of the aortic arch. The patient's transcranial Doppler (TCD) waveform was flat throughout all segments of the intracranial arteries. Intravenous acetazolamide injection confirmed the severe impairment of vasoreactivity. After a bypass graft from the aorta to the left CCA, flow velocity and pulsatility were dramatically increased without postoperative complications. A nonpulsatile cerebral perfusion indicates severe hemodynamic impairment and is partially reversible by a surgical bypass graft. TCD seems to be useful to detect "high-risk" patients and to follow up in TA. PMID- 12722502 TI - Harmonic imaging in acute stroke: detection of a cerebral perfusion deficit with ultrasound and perfusion MRI. AB - Perfusion harmonic imaging of the brain is a new ultrasound technique for assessing cerebral perfusion. In a patient with acute middle cerebral artery infarction, this method detected a cerebral perfusion deficit corresponding to the area of delayed perfusion as displayed by perfusion magnetic resonance imaging. PMID- 12722504 TI - [Guarding the interests of justice]. AB - The article is devoted to the history of creation and formation of forensic medical examination in the Red Army. The difficulty in formation and topical of activity of army, front and district forensic medical laboratories are emphasized. The orderly structure of expert institutions created during the years of the Great Patriotic War allowed to lay the scientific and methodical foundation of activity of the forensic medical examination in the Armed Forces. Special contribution into creation and organization of military expert service was made by Mickail Ivanovitch Avdeyev--the chief expert of the Red Army. The peculiarities of expert activity and its significance both during the years of the Great Patriotic War and today in difficult conditions of the Russian Federation Armed Forces reformation are discussed in terms of the history. PMID- 12722505 TI - [Scientific investigations conducted in forensic medical examination]. AB - The authors discuss the conceptual problems connected with necessity to reform the research activities in the RF MD expert institutions within the "Program of development of RF MD forensic medical examination for the period of 2001-2005", the character of its main directions. The complex of concrete research and experimental works is described. PMID- 12722508 TI - [Heart failure in casualties with explosive mine trauma]. PMID- 12722506 TI - [On the state of psychiatric services in the Armed Forces of the Russian Federation]. PMID- 12722507 TI - [Organization of work in the North-Caucasus region for the admission, investigation and shipment of killed servicemen]. PMID- 12722509 TI - [Microcirculatory disorders caused by gunshot wounds of the small intestine (experimental study)]. AB - The mirocirculatory dynamics in intestinal wall at different distances from the wound canal was established in experimental dogs using the stimulation of jejunal high-energetic gunshot injury. The radial spectrum of microcirculatory changes is characterized by the following parameters: the decreased arterio- and arteriole spasm, the increased insufficiency of venous outflow, the capillary net "thickening" and the increased load on lymph drainage system. Temporal dynamics is manifested by insignificant resolution of arterio- and arteriole spasm with remained microcirculatory disorders. Together with rather stable microvascular morphometric parameters there were the following disorders: the irregular spasm and sludge in capillaries, the endotheliocyte swelling, the increased venous congestion. The microcirculatory state determines both the degree of morphologic changes in the zone of gunshot injury of intestinal wall and the intensity of compensatory-and-adoptive processes. PMID- 12722511 TI - [Prognostic significance of arterial hypertension in the formation of hypertensive disease in youths]. PMID- 12722510 TI - [Improvement of the rehabilitation system of patients after aortocoronary bypass]. AB - Using the significant representative sample the important conclusion was drawn about the necessity of patient early differentiation on the base of psychosomatic status evaluation. The algorithms of treatment measures were developed on the base of which two rehabilitation patterns were created. It is shown that the use of the patterns developed permits to decrease the treatment expenses (the important factor in modern conditions). For practical work the new integral indices are proposed that allow to improve the diagnosis quality and to work out the tactics of individual restorative treatment. The necessity to improve the medical rehabilitation system using the differential approach to determination of need of restorative treatment and place of its conduction is proved. PMID- 12722512 TI - [On the issue of the diagnosis and correction of disease-related malnutrition]. PMID- 12722513 TI - [Low-dose digital roentgenography of the thoracic cavity: comparative evaluation of the method]. PMID- 12722514 TI - [Effect of chemoprophylaxis on pneumonia incidence in organized groups]. PMID- 12722515 TI - [Pathology and chemical verification and detection of "risk group" of endogenous psychosis]. PMID- 12722516 TI - [Botulism in servicemen: diagnosis and treatment]. PMID- 12722517 TI - [Women serving in the Russian Army]. PMID- 12722518 TI - [Women--military medical paralegals with forensic medical expertise]. PMID- 12722519 TI - [N M Aleksandrov--an outstanding Russian maxillofacial surgeon]. PMID- 12722521 TI - Arthroscopic monopolar radiofrequency thermal capsulorrhaphy for the treatment of shoulder instability: a prospective outcome study with mean 2-year follow-up. AB - PURPOSE: This study describes the experience with arthroscopic monopolar thermal capsulorrhaphy for painful multi-directional shoulder instability in a community based practice. TYPE OF STUDY: Prospective outcome study of consecutive patients. MATERIALS AND METHODS: Thirty-two shoulders in 30 patients were prospectively evaluated using the twelve-question Simple Shoulder Test (SST). This questionnaire assesses shoulder pain and function in a variety of everyday tasks. The duration of symptoms prior to surgery averaged 26.1 +/- 41.0 months (range 2 months-20 years). Patients were evaluated at baseline and then again at various follow-up time points (mean follow-up of 27.7 +/- 8.6 months post-surgery, range, 4-41 months). Patients completed at least 4-6 weeks of physical therapy prior to surgery. All patients were determined to have multi-directional instability during examination under anesthesia and there were no Bankart lesions in the series. When thermal capsulorrhaphy alone was determined to not be adequate to achieve a stable shoulder, the procedure was augmented with additional tuck sutures in whatever direction necessary. Twenty-one of the 32 surgeries (62%) used suture capsulorraphy in addition to the thermal treatment. Following surgery patients were immobilized in a sling for a minimum of 4 weeks and were not allowed to return to sports or strenuous activity until 12 weeks postoperatively. RESULTS: Mean pre-operative SST score was 5.5 +/- 3.5. Post-operative mean SST score was 9.0 +/- 3.3. This difference was significant (p < 0.0001). Five patients in the series were unable to return to activities of daily living and were considered failures. Two of these patients received a second operation that was then successful. CONCLUSIONS: Arthroscopic monopolar radiofrequency thermal capsulorrhaphy alone in or addition to suture capsular tightening procedures is a successful means of treating patients with painful, symptomatic, multi directional instability. PMID- 12722522 TI - Child head injuries: review of pattern from abusive and unintentional causes resulting in hospitalization. AB - OBJECTIVE: Comparison of patterns of hospitalized child head injuries among unintentionally injured with intentional Shaken Baby Syndrome and Abused victims. METHODS: Medical records of children birth to 4 years of age admitted to the Fletcher Allen Health Care Hospital in the years 1993 to 1999 for head injury due to any cause were reviewed. Reviews which included age, gender, site of injury, caretaker of child, mechanism of injury, time of injury, severity of injury, CNS sequelae, and quality of investigation were completed for each child. RESULTS: Of the total of 85 records reviewed, 49 were male and 36 female with a mean age of 18.9 months. Seventy-three children were injured unintentionally. Twelve were victims of intentional actions. Fifty-three percent of the unintentionally injured were male and 83% of the abused were male. Falls caused 53%, motor vehicles 17%, abuse 14%, of all the 85 hospitalized children. Only three deaths occurred among the 85 children, all from motor vehicle crashes. Forty-two percent of the 12 abused victims suffered serious CNS injury compared to only 10% among the unintentionally injured. Earlier symptoms and signs of abuse were missed in four of the 12 abused children. CONCLUSIONS: Demographic patterns of children hospitalized in Vermont for head injuries are similar to other state and national studies. Severity of injury is significantly higher for abused children. Primary health care providers should receive training emphasizing higher diagnostic index of suspicion for abusive head injuries. PMID- 12722523 TI - A qualitative study of the importance and etiology of chronic respiratory disease in Alaska native children. AB - OBJECTIVES: To generate new hypotheses about factors that may contribute to chronic respiratory disease in Alaska Native children in rural Alaska. METHODS: Qualitative formative research with interviews of community members, village healthcare providers, and referral providers in the Yukon-Kuskokwim Delta Region of Alaska. RESULTS: The respondents identified chronic and acute respiratory illnesses as the most important serious child health problems of the region. They believed that chronic respiratory conditions, especially asthma, were increasing. The most frequently discussed potential contributing factors were smoke, dust, feeding practices, socioeconomic conditions, and mold. The intervention mentioned most frequently that would make the greatest impact was a more timely diagnosis of asthma. CONCLUSIONS: Reports of an increase in chronic respiratory problems are parallel with reports of increases in dust and mold in the communities. Further environmental quantitative research is necessary to confirm the importance of these factors. In addition, improved education for identification and treatment of asthma would appear to be beneficial. PMID- 12722524 TI - [Consumer protection or self-interest?]. PMID- 12722525 TI - [Antiretroviral treatment of HIV infected patients]. PMID- 12722526 TI - [Autologous chondrocyte transplantation]. PMID- 12722527 TI - [Motion sickness. Findings and treatment]. PMID- 12722528 TI - [Unexplained vague abdominal complaints in adults: indication for CT in case of suspected intussusception]. AB - In two adult patients, a 74-year-old woman and a 84-year-old man, who suffered from vague abdominal complaints, an intussusception was diagnosed by CT. Surgical resection of the affected bowel parts was successful. Intussusception is usually seen in children; in adults it is a rare condition. Adult patients mostly complain about vague abdominal pain only. Physical examination, laboratory investigations and plain abdominal X-rays often don't give any additional information. In such patients it is advised to perform CT of the abdomen at an early stage. CT may show a so-called 'target sign' which is characteristic of an intussusception. In addition it can provide information about the possible causes of the intussusception, most commonly a malignant tumour in adults. During laparotomy one should not attempt to reduce the intussusception because of the risk of tumour spill. In this clinical review, we present two adult patients with unexplained abdominal complaints due to intussusception, caused by malignancy. PMID- 12722529 TI - [Treating hypertension is more important than the choice of agent; results from the largest clinical trial until now]. AB - During the past few years, several randomised trials have compared the effects of older blood-pressure lowering drugs (diuretics, beta-blockers) with those of newer ones (angiotensin converting enzyme (ACE) inhibitors, calcium entry blockers) on the long-term prognosis. In general, no significant differences were found between these regimes. Recently, the ALLHAT trial, which was the largest hypertension trial ever and in which over 40,000 patients with hypertension participated, was completed. The initial treatment consisted of either the diuretic chlorthalidone, the calcium entry blocker amlodipine, the ACE inhibitor lisinopril, or the alpha-blocker doxazosin. The latter arm was prematurely discontinued because of a higher incidence of the secondary endpoint heart failure and stroke. Based on an intention-to-treat analysis, the other types of treatment proved to be equivalent in terms of the primary endpoint, a composite of fatal coronary heart disease and non-fatal myocardial infarction. Although the investigators conclude that ALLHAT suggests that thiazide diuretics should be first choice in the treatment of hypertension, there are several caveats that tend to lessen the strength of this conclusion. PMID- 12722530 TI - [Current validation of the Greulich and Pyle atlas for the determination of skeletal age]. AB - The radiological atlas of Greulich and Pyle published in 1959 has been validated for the modern measurement of skeletal development in a population of white school-age children from Rotterdam. This is a useful study result. Although contemporary children grow to become much taller than in previous generations, this increased growth potential is apparently realised within the same time span. PMID- 12722531 TI - [New developments in the treatment of nausea and vomiting caused by chemotherapy]. AB - With the advent of the 5HT3-receptor antagonists in the 1990s and their combination with dexamethasone, complete emesis protection during the first 24 hours after administration of an emetogenic cytostatic agent became possible in 70% of the patients. Despite acute emesis protection through the use of 5HT3 receptor antagonists and dexamethasone, 40% of the patients do have symptoms during the following days. 5HT3-receptor antagonists and dexamethasone are modestly effective in this delayed phase and the antiemetic protection decreases progressively with multiple cycles. Neurokinine(NK)-1-receptor antagonists belong to a new class of antiemetic agents that specifically target the NK-1-receptor which is involved in both the acute, and in particular, the delayed phase. Clinical studies have demonstrated that the addition of the NK-1-receptor antagonist apprepitant to a 5HT3-receptor antagonist plus dexamethasone combination improves acute emesis protection by about 20% (from approximately 60% to 80%) and by about 30-40% in the delayed phase (from approximately 30% to 60 70%). The effectiveness of this triple therapy is sustained during subsequent cycles. The chance that an individual patient will be completely protected from severe nausea and vomiting during the entire course of chemotherapy (often 6 cycles) consequently increases significantly (from 34% in the placebo group to 59% in the aprepitant group). PMID- 12722532 TI - [Dutch Institute for Health Care Improvement revised guideline, 'Sexually transmitted diseases and neonatal herpes']. AB - The Dutch Institute for Health Care Improvement revised guideline, 'Sexually transmitted diseases and neonatal herpes' summarises the current scientific position on the diagnosis and treatment of a great number of sexually transmitted diseases (STD) and neonatal herpes. Symptomatic treatment of suspected Chlamydia trachomatis infection and gonorrhoea without previous diagnosis is not recommended. Treatment can be started immediately, once samples have been taken. Risk groups eligible for screening or proactive testing on C. trachomatis infection include: partners of C. trachomatis-positive persons, visitors of STD clinics, women who will undergo an abortion, mothers of newborns with conjunctivitis or pneumonitis, young persons of Surinam or Antillean descent, young women with new relationships and individuals whose history indicates risky sexual behaviour. A period of 3 months can be adopted between a risky contact and the HIV test (this used to be 6 months), unless post-exposure prophylaxis was used. For the treatment of early syphilis no distinction is drawn between HIV infected and non-HIV-infected persons. It is no longer recommended that women in labour with a history of genital herpes are tested for the herpes simplex virus. Virological testing of the neonate is only advised if the mother shows signs of genital herpes during delivery. PMID- 12722534 TI - [Risk of ventricular fibrillation in patients with Wolff-Parkinson White syndrome]. AB - A 16-year-old boy suddenly fell off his stool, a 26-year-old man had persistent palpitations and a 29-year-old man was reanimated without an incriminating anamnesis. The diagnosis 'Wolff-Parkinson-White(WPW)-syndrome' was made in all three cases. The boy died as a result of postanoxic neurological injury; in the two men, further cardiac rhythm disturbances were prevented by interrupting the accessory atrioventricular connection via radiofrequency catheter ablation. In ECG databases, a WPW-pattern is encountered in 1-3 of 1000 electrocardiograms. Atrial fibrillation with 1:1 conduction via the accessory pathway, leading to ventricular fibrillation, is the most common cause of sudden death in WPW patients. In some cases, atrial fibrillation with a rapid ventricular response is the first sign of the syndrome. The risk of sudden death in these patients is estimated to be 0.0-0.6% per patient per year and cannot be predicted easily. Curative treatment is possible in the form of radiofrequency catheter ablation. PMID- 12722533 TI - [Diagnostic image (134). A woman with a crepitating arm. Post trauma emphysema]. AB - A 35-year-old woman presented with extensive subcutaneous emphysema caused by trapping of air by valve-like wound edges after a minor trauma of her right olecranon. PMID- 12722535 TI - [Two patients with a perforation of the esophagus and hypopharynx, respectively, caused by a bone in their food]. AB - Two patients, men aged 58 and 28 years, developed pain in the throat while eating chicken and pork, respectively. In the emergency clinic and during a check-up on the following day, no foreign body could be found. On the third visit to the hospital, oesophagoscopy was performed. In one patient a chicken bone was found lying transversely in the oesophagus; after removal of this bone extraluminal pus was seen. In the other patient a pork bone was found in the right piriform recess, with a great deal of local pus. Both foreign bodies were removed during oesophagoscopy. The recovery of both patients was uneventful. A foreign body is found in the upper digestive tract in approximately one-third of the patients presenting to an emergency department with a history of foreign body ingestion. Serious complications occur in less than 1%; however, these complications can be life-threatening. If, based on the anamnesis, physical examination or radiological evaluation, there is suspicion of a foreign body in the oesophagus, or if the examination is incomplete, then oesophagoscopy should be performed. PMID- 12722536 TI - [A degradable artificial nerve guide to bridge peripheral nerve defects]. AB - The standard technique for bridging a peripheral nerve defect is an autologous nerve graft if the nerve ends cannot be sutured. Recent evidence indicates that an alternative procedure-application of a degradable nerve guide-may be feasible. Currently the use of such a degradable nerve guide for the recovery of peripheral nerve defects in the hand is being tested in a multicenter trial. Conventional suturing as well as autologous nerve grafting are accepted methods in the control group within the protocol of the multicenter study. In the first two patients to receive the implant, a 28-year-old man and a 50-year-old women with tendon and nerve injury due to glass cuts, the operation was technically successful. Functional nerve recovery will be assessed in due course. PMID- 12722537 TI - [Early diagnosis of rheumatoid arthritis with a test based upon a specific antigen: cyclic citrullinated peptide]. PMID- 12722538 TI - [The small epidemiologic transition: further decrease in infant mortality due to medical intervention during pregnancy and childbirth, yet no decrease in childhood disabilities]. PMID- 12722539 TI - [Quality assessment of medical-scientific research: in the future it should also be assessed on the basis of social impact]. PMID- 12722540 TI - [New active agents approved as pharmaceutical specialties in 2002]. PMID- 12722541 TI - [Pharmaceutical care in the aged: illustration of the process in patients hospitalized in Canada]. PMID- 12722542 TI - [St. Mary's Thistle: an overview]. AB - St. Marys Thistle has been approved for registration as a regular medicine in Belgium. The hepatotropic properties of this plant are rather difficult to evaluate objectively. Mortality rate in case of life-threatening hepatic diseases is the most objective parameter. Legalon is the only drug registered in Belgium. It has a prescription only status. The plant Silybum marianum is a thistle and as a consequence belongs to the Compositae. There is a limited production of St. Marys Thistle in Pajottenland, west of Brussels. The seeds are exported to Italy in order to extract silymarine, a mixture of flavonolignanes with antioxidant properties. Silymarine has been tested in living animals deliberately intoxicated with mushroom toxins, medicines, heavy metals or toxic organic solvents. Preventive as well as curative activity has been confirmed. Silymarine accumulates in the liver, which is also the target organ in therapy. Silymarine improves the prognosis after accidental ingestion of the toxic Amanita phalloides. Patients infected with hepatitis B and C might benefit from Silymarine, but more data have to be generated. Silymarine given to patients with liver damages by alcohol lowers the death toll. The drug has a general safety pattern comparable to placebo. PMID- 12722543 TI - [Can we neutralize skin aging?]. PMID- 12722544 TI - [Drug eluting stents, a priceless advance!]. PMID- 12722545 TI - [Use of coronary stents]. PMID- 12722546 TI - [Comparison between radial and femoral approaches in ad hoc coronary angioplasty]. AB - Between September 1999 and June 2001, 591 patients required ad hoc coronary angioplasty. The authors compared the group of patients in whom the approach of first intent was radial (n = 328, 55%) with those in whom the femoral approach had been used (n = 263), in terms of immediate local (haematoma or thrombosis requiring surgical intervention or transfusion) and general complications (myocardial infarction, cerebrovascular accident), and major adverse cardiovascular events (infarction, angioplasty, bypass and death) at 1 year. The dose of ionising radiation during the procedures was also compared prospectively. The conversion rate from the radial (R) to the humeral or femoral (F) approach was 10%. The angioplasty, stenting, and stenting without dilatation failure rates were identical in the two groups (5% versus 5%, 0.6% versus 1.9%, 3% versus 4%, respectively). The average irradiation time was greater in the R group than in the F group (23 +/- 12 min vs 17 +/- 4 min, p < 0.001) as was the irradiation per surface unit (242 +/- 137 Gy.cm2 vs 185 +/- 117 Gy.cm2, p < 0.001). The immediate complication rate was comparable in the two groups (2.5% in group R vs 3.6% in group F) as was the major adverse cardiovascular event rate at 1 year (13% in both groups). The authors observed the same rate of immediate complications and late adverse cardiac events in patients undergoing coronary angioplasty followed by immediate angioplasty by the radial or femoral approaches with an acceptable conversion rate from the radial to the femoral approach. The procedures by the radial approach seem to be associated with a greater time and dosage of ionising radiation. PMID- 12722547 TI - [Utility of BNP measurement in the emergency room in patients with suspected unstable angina with a normal ECG]. AB - Unstable angina is a serious condition, difficult to diagnose in the emergency room. Clinical, electrocardiographic and biological signs (increased troponine) are not sensitive. The authors set out to assess whether measuring B natiuretic peptide in the emergency room was more sensitive for identifying symptomatic coronary lesions. One hundred and twenty patients admitted to the emergency room for chest pain compatible with the diagnosis of unstable angina and a normal ECG were included in this prospective study. All patients underwent coronary angiography during their hospital admission. The sensitivities of troponine at a threshold of 0.4 ng/ml and of brain natiuretic peptide (BNP) at a threshold of 10 pg/ml in this population were 66% and 92% respectively. The use of troponine and BNP together provided better results than troponine and BNP alone for the identification of patients with chest pain with significant coronary lesions. PMID- 12722548 TI - [The future of the cardiologist: job and professional conditions]. PMID- 12722549 TI - [Economic repercussions of cardiac insufficiency in France]. AB - The aim of the article is to review economic and public health consequences of congestive heart failure in France. This disease with an increasing prevalence induces for the social security system and the society a high cost; 85 to 93% of this cost are bound to hospitalizations, most of them avoidable. New modalities of care are to be chosen in order to improve the management of heart failure and to contain costs. In this context, multidisciplinary interventions based on patients' education are experimented taking care to assure a greater role to health actors working in the ambulatory sector (GPs, private cardiologists, nurses). PMID- 12722550 TI - Impact of genetic polymorphisms on heart failure prognosis. AB - Recent progress in genomic applications have led to a better understanding of the relationship between genetic background and cardiovascular diseases such as heart failure. The broad variability in heart failure patient outcome is in part secondary to modifier genes, i.e. genes that are not involved in the genesis of a disease but modify the severity of the phenotypic expression once the disease has developed. The strategy most commonly used to identify modifier genes is based on association studies between the severity of the phenotype and the sequence variation(s) of selected candidate gene(s). Using this strategy, several polymorphisms of the beta 1 and beta 2-adrenergic receptors genes and the angiotensin converting enzyme gene have been correlated to the prognosis of patients with heart failure. Recently, we have applied an experimental strategy, known as genome mapping, for the identification of heart failure modifier genes. Genome mapping has previously been used with success to identify the genes involved in the development of both monogenic and multifactorial diseases. We have shown that the prognosis of heart failure mice, induced through calsequestrin overexpression, is linked to two Quantitative Trait Loci localized on chromosomes 2 and 3. Using both strategies (candidate gene and genome mapping) should allow us to identify a number of modifier genes that may provide a more rational approach to identify patients with the worst prognosis and to predict their response to therapy. PMID- 12722551 TI - Statins: maid-of-all-work in cardiovascular diseases! AB - Inhibitors of HMG-CoA reductase (statins) lower the level of circulating LDL-C by blocking the activity of HMG-CoA reductase. Their efficiency to prevent cardiovascular events was demonstrated in several clinical trials for primary and secondary prevention. However, subgroups analysis of trials together with experimental studies have increasingly documented that the beneficial effects of statins extend beyond the sole reduction in LDL-C. These effects include improvements of vasoreactivity, haemostasis and plaque stability, reduction of pro-inflammatory events such as a decrease in monocyte adhesion and infiltration, oxidation level and pericellular proteolysis. Possible repair of ischemic tissues through enhancement of mobilization of endothelial progenitor cells are also described, although more investigation are needed to clearly identify the role and safety of statins in angiogenesis. These pleiotropic effects are generally explained by the fact that statins inhibit the intracellular production of metabolites located downstream of mevalonate in the cholesterol pathway, such as isoprenoids (farnesyl pyrophosphate and geranylgeranylpyrophosphate). These hydrophobic metabolites allow the membrane anchorage of small G proteins (Ras and Rho) as well as the Gy subunit of heterotrimeric G proteins, a post-translational step that is critical in the regulation of G protein signaling activity. These drugs are therefore a valuable tool not only for the clinician but also for the biologist, allowing to investigate the regulation of gene expression that is controlled by the intracellular activity of membrane-anchored prenylated signaling proteins. PMID- 12722552 TI - Oxidative stress and heart failure. AB - Despite advances in treatment, chronic congestive heart failure carries a poor prognosis and remains a leading cause of cardiovascular death. Accumulating evidence suggests that reactive oxygen species (ROS) play an important role in the development and progression of heart failure, regardless of the etiology. Under pathophysiological conditions, ROS have the potential to cause cellular damage and dysfunction. Whether the effects are beneficial or harmful will depend upon site, source and amount of ROS produced, and the overall redox status of the cell. All cardiovascular cell types are capable of producing ROS, and the major enzymatic sources in heart failure are mitochondria, xanthine oxidases and the nonphagocytic NADPH oxidases (Noxs). As well as direct effects on cellular enzymatic and protein function, ROS have been implicated in the development of agonist-induced cardiac hypertrophy, cardiomyocyte apoptosis and remodelling of the failing myocardium. These alterations in phenotype are driven by redox sensitive gene expression, and in this way ROS may act a potent intracellular second messengers. Recent experimental studies have suggested a possible causal role for increased ROS in the development of contractile dysfunction following myocardial infarction and pressure overload, however the precise contribution of different cellular and enzymatic sources involved remain under investigation. PMID- 12722553 TI - [Recommendations of the French Society of Cardiology concerning indications for doppler echocardiography]. PMID- 12722554 TI - [Opening of the XIIIth European meeting of the French Society of Cardiology]. PMID- 12722555 TI - [Case reports of suicide with an electric drill]. AB - In this report, three cases of suicide by an electric power drill are described. In the first case, a man aged 51 was found with four penetrating wounds of the head and chest. The second case involves a 56-year-old man who suffered a penetrating craniocerebral injury and the third case involves a 53-year-old man who suffered one head injury and two chest injuries. In the first two cases, the victims were found dead. In the third case, the victim survived his injuries for 28 hours. The case histories and the post-mortem examinations revealed no evidence of homicide. PMID- 12722556 TI - [Fatal outcome of Ecstasy overdose]. AB - Consumption of amphetamine derivatives has considerably increased in Germany since the early nineties. Again and again intoxications with lethal outcome have also been reported, especially after physical activities such as intensive dancing. The authors present a case of an obviously suicidal intoxication of a 21 year-old man who was found dead with marked cuts on the right forearm. Toxicological tests showed in particular 3, 4-methylene-dioxymethamphetamine (MDMA). The results of the hair analysis revealed chronic consumption, but no cellular liver damage could be demonstrated. When examining the body fluids and organs, the highest concentrations by far were measured in the lungs (36.6 mg/kg), the liver (29.7 mg/kg) and the brain (29.1 mg/kg). The concentration in heart blood amounted to 10.8 mg/kg and was thus markedly higher than in peripheral blood (7.2 mg/kg). In the muscles concentrations ranged between 14.3 mg/kg and 20.2 mg/kg. On the basis of these concentrations and the available pharmacokinetic data the amount of MDMA probably consumed is assessed. It is demonstrated that for this assessment the concentrations in the muscular system are of special importance, as redistribution of highly lipophilic substances from the surrounding tissue is possible also in peripheral blood. PMID- 12722557 TI - [An unusual case of double death]. AB - The report presents the case of an alleged double homicide or a homicide followed by suicide of a couple, in which the court-ordered autopsy of both victims revealed that the homicide had been committed by the husband immediately before he died himself from cardiac failure. The article gives a short summary of the pathophysiological relations between emotional tension (stress) and physical strain and the effects on the cardiovascular system. The case report describes a homicide by strangulation immediately followed by the perpetuator's death from a natural cause. PMID- 12722558 TI - [Mixed fatal poisoning caused by taking L-methadone and chloral hydrate]. AB - A 42-year-old female drug user who was enrolled in a methadone maintenance program was found dead in her apartment. Cause of death was an intoxication with chloral hydrate and L-methadone. Trichloroethanol (TCE), the primary metabolite of chloral hydrate, was quantified by solid phase microextraction (SPME) and GC/MS in heartblood (27 micrograms/ml) and urine (338 micrograms/ml). D- and L methadone were differentiated by chiral HPLC, which showed that only L-methadone had been taken. The quantitation of L-methadone and its metabolite EDDP was carried out by GC/MS from heartblood (1300 ng/ml and 86 ng/ml, respectively), urine (5239 ng/ml and 4960 ng/ml, respectively) and gastric contents (159 ng/ml and 122 ng/ml, respectively). The concentrations of both--trichloroethanol and methadone--were in toxic ranges. PMID- 12722559 TI - [Heteroplasmy in mtDNA in a set of dizygotic twins]. AB - The authors report on a case of a dicygotic female twin pair, one of them containing a T/C heteroplasmy in two (heart and brain) of five analysed organs at position nt204 of the HV2 region. Results and especially the evidence of an unequal distribution of the heteroplasmy are discussed. PMID- 12722561 TI - Home health quality goes public. PMID- 12722560 TI - [Causality of an accident in subsequent fatal pulmonary embolism a few hours after trauma]. AB - An 87-year-old woman, who was largely immobilized in a wheelchair, suffered rib fractures and an unstable fracture of the pelvic ring in a fall. Approximately 2 1/2 hours later she developed marked clinical symptoms of pulmonary thromboembolism and died 5 hours after the accident. The question to be discussed with regard to the causality was the unusually short interval of only 2 1/2 to 5 hours between the accident and the clinical signs of embolism leading to her death. Current literature gives only rough outlines stating that thromboembolism can occur as early as the first day after the trauma. An alternative theory explaining the pulmonary thromboembolism may be the breaking off of a pre existing thrombus due to manipulation during transport or diagnostic measures in connection with the unstable fracture of the pelvic ring. PMID- 12722562 TI - Winning the race against the nursing shortage. PMID- 12722563 TI - Living with stress since 9/11. PMID- 12722564 TI - Charting the biologic frontier. Interview by Val J. Halamandaris. PMID- 12722565 TI - Caregiving. PMID- 12722566 TI - A tribute to Congressman W.J. "Billy" Tauzin. PMID- 12722567 TI - How to use the hidden asset of testimonials. PMID- 12722568 TI - The challenge of balancing staff and patient needs. PMID- 12722569 TI - State budget deficits connected to Medicare cuts in home care. PMID- 12722570 TI - Immediate approval of health plan participant's cancer treatment ordered. Czarnopys v. Crystal Flash Ltd. Partnership of Michigan. PMID- 12722571 TI - Health education and health promotion--implementing healthy public policies. PMID- 12722572 TI - Promoting sexual health to young people: preventing teenage pregnancy and sexually transmitted infections. PMID- 12722573 TI - Unprotected sexual intercourse in teenagers--causes and consequences. PMID- 12722574 TI - Increases in sexually transmitted infections--a challenge for sexual health promotion. PMID- 12722575 TI - Pre-eclampsia: prediction. PMID- 12722577 TI - A new year, a new commission: the commission for patient and public involvement in health. PMID- 12722576 TI - Pre-eclampsia: prevention. PMID- 12722578 TI - Health information on the Internet. PMID- 12722579 TI - Levelling the playing fields of England: promoting health in deprived communities. AB - Health promotion is about focusing on systems in which those 'waiting to be ill' are living their lives now. Unlike the old, the new public health agenda is about developing and implementing healthy public policies. This stance involves everyone working together to develop change in the conditions of living. This article presents a brief overview of the role of the health promotion specialist in a Primary Care Trust working with six Sure Start Programmes in a town that has high levels of poverty and deprivation. To demonstrate the principles of 'health for all', the article concludes with an overview of the development of a partnership to promote initiation and continued exclusive breastfeeding using collaborative working, community participation and equity. PMID- 12722580 TI - A review of social housing regeneration in the London Borough of Brent. AB - Council high-rise estates sprang up rapidly during the 1960s and 1970s, with cross-governmental support to resolve the nation's housing crisis. It soon became apparent that many such new estates, designed by remote architects, and sometimes constructed rapidly and unsatisfactorily, did not provide the ideal living initially perceived. Many estates had early problems with architecture, construction and design. They proved an inhumane environment for many residents and there were frequent problems with communal features. Such estates were soon stigmatised and difficult to let as increasingly residual households were placed there, creating majority welfare-dependent estates. This created a downward spiral that traditional, and remote, housing management found difficult to address, and was too wide-scale to rectify financially within existing regimes. Prior to the 1980s, there were no specific government policies to tackle housing regeneration on high-rise estates. Problems became wider than traditional housing management and poor housing environments, encompassing social and economic exclusion. This paper, based on historical and contemporary literature as well as estate visits, reviews regeneration policy in three council housing estates within the London Borough of Brent. It traces successive government approaches since the 1980s from one that challenged the very status of council housing- notably at Stonebridge Park and Chalkhill, to one of partnership with the local authority--at South Kilburn. Housing policy is now concerned with more than just housing--it is about moving toward social inclusion, which requires initiative, flair, resource and commitment. It is about new accountabilities--not just numbers of bricks and mortar constructions, but about the lives, opportunities and health of those who live in an area. This fundamentally involves a partnership approach with residents at the centre of regeneration. This paper finds that sustainable estate regeneration policies are about continued improvements in both housing policy and social development. PMID- 12722581 TI - Mixed signals from Kerala's improving health status. AB - It can be said that health policy is about creating the conditions to free a population from disease and impairment, and so prolong the quality of life for all. If this is its purpose, then the practice of public health policy in the Indian state of Kerala presents a conundrum for health promotion analysts. On the one hand, Kerala has had remarkable success in reducing its infant mortality rates to around one-third of the Indian average, but on the other, its morbidity rates have risen to impose an economic cost more than three times the national level. This paper sets out to explain this apparent contradiction in outcomes, serving both to provide a review of Kerala's health status and as an illustrative guide for the interpretation of the effects of health policy in some other developing areas. While the essential health data for Kerala might imply an unsatisfactory future, the authors argue that the rapid improvements that have occurred simultaneously in public education are more likely to explain an increase in the effective reporting of disease conditions, thus suggesting a more optimistic scenario for future practice that can lead to a sustained improvement in health care. PMID- 12722582 TI - The survival and recovery of bacteria in vacuum cleaner dust. AB - The possibility exists that environmental dust could be a source of gastro intestinal infection in the domestic environment and that the causative microbes are collected during vacuum cleaning. This study examines the survival of total bacterial populations, Enterobacteriaceae and salmonella species in vacuum cleaner dust in vitro and in use. Total counts remain constant at around 10(6) 10(8) colony forming units (cfu) g-1 for at least 60 days. Enterobacteriaceae showed only a slight decline over the same period. Recovery of salmonellae artificially inoculated into vacuum dust was dose and time dependent: even relatively small inocula (< 100 cfu g-1) were recoverable more than one month after inoculation. Questionnaires and vacuum cleaner dust samples were received from 76 households. A significant association was found between total bacterial counts and whether the household was in a rural or urban situation and the number of people in the household. No association was found between total bacterial counts and the number of children in the household, the presence of pets, wearing shoes indoors, the makes, model and age of the vacuum cleaner, the type of dust collector or dust bag, how often the vacuum cleaner was used and the length of time between sample collection and microbiological testing. Salmonella species were isolated from vacuum cleaner dust from three of the 76 households although no factors could be identified linking the homes. This study shows that vacuum cleaners are effective collectors and reservoirs of microbial contamination and that these contaminants are able to survive for up to two months. Vacuum cleaner dust could thus be a useful indicator of environmental contamination in the home. PMID- 12722583 TI - Reducing domestic exposure to environmental tobacco smoke: a review of attitudes and behaviours. AB - This paper reviews research on attitudes and behaviours towards environmental tobacco smoke (ETS), with a special focus on child health and the indoor environment. Research needs and ways forward to encourage reductions in domestic ETS levels are discussed. Published material was identified through online literature searches (Medline, Toxline, Cancerlit, Biosis, Embase, Enviroline, Sociological Abstracts, Social Science Citation Index, Academic Index and Psychinfo). The literature search strategy employed search terms such as "passive smoking" or "environmental tobacco smoke" with "attitude" or "awareness" and other synonyms. Additional publications were identified by citation chasing and expert advice. Focusing on the UK, studies that provided survey-derived data about attitudes and behaviours in relation to ETS exposure in the indoor environment were selected for review. Published studies from other countries were also included when they provided information pertinent to this review. Most people are aware of the health risks associated with ETS exposure, and there is a high level of support for smoking restrictions in public places to protect non smokers from ETS. However, although there is concern among both non-smoking and smoking parents about children and second-hand smoke, many people allow children to be exposed to ETS in the home. The review suggests that traditional health promotion campaigns have had only limited success in encouraging ETS risk reduction measures in the home. Because ETS is a public health priority, particularly in relation to child health, the barriers to the uptake of such measures need to be explored in detail to inform the future promotion of reductions in domestic levels of ETS. PMID- 12722584 TI - Lessons to be learned: a case study approach. Underage drinking in adolescents. AB - The drinking habits of children aged 11 to 16 years attending a comprehensive school are reported. Pupils were able to access alcoholic drinks readily despite being underage. A general awareness of the health and social risks of drinking did not act as a deterrent and, alarmingly, only less than 10% of the group reported that they did not drink. Health education and the role that schools can play are highlighted. PMID- 12722585 TI - Historical perspectives on health. The Parkes Museum of Hygiene and the Sanitary Institute. AB - The Parkes Museum was founded in 1876 to commemorate the life and work of Edmund Alexander Parkes (1819-1876), who was the first Professor of Military Hygiene at the Army Medical School and one of the pioneers of the public health reforms of the nineteenth century. The Museum was a multi-disciplinary institution which aimed to teach an awareness of public health matters to the general public and members of the building trade, thereby encouraging healthy design of living accommodation. The Sanitary Institute had been founded in the same year and sought to improve public health practice by setting standards for health professionals. The aims of the two organisations were therefore complementary. From 1883 they occupied the same premises in Margaret Street, London, and in 1888 they amalgamated to form a joint organisation known as The Sanitary Institute. Despite a move of location in 1909, the Parkes Museum continued to exist until the mid-1950s when an extensive refurbishment, together with a change of focus from museum to exhibition, led to its reopening in 1961 as the Health Exhibition Centre. It closed in 1971 when The Royal Society for the Promotion of Health, the successor to The Sanitary Institute, moved to new accommodation in Grosvenor Place, London. PMID- 12722586 TI - [Application of the criterion of biological time (tau n/tau 0) to the studies of embryogenesis in birds]. AB - It was shown on the example of chick embryo that the number tau 0 (tau n/tau 0) can be recommended as a measure of biological time and, for this purpose, the duration of the minimal mitotic cycle during synchronous cleavage divisions should be determined (in minutes) in various avian species. Based on the preliminary data, one can propose the comparability and similarity of the temporal programs of gastrulation and somitogenesis in the chick embryo and embryos of some fish and amphibians. PMID- 12722588 TI - [Application of the model system of hormonal stimulation of the sturgeon oocyte maturation and ovulation in vitro for solving some fundamental and applied problems]. AB - The author summarizes the results of many-year application of the model of in vitro sturgeon oocyte maturation for different purposes, such as comparison of gonadotropic activities of different preparations, selection of females for breeding, and studying the effects of different factors in order to improve the breeding technology. Special attention is paid to factors that can affect the results of experiments on hormonal stimulation of in vitro oocyte maturation and ovulation and their interpretation. Two other phenomena are discussed: the inhibitory effect of gonadotropic pituitary hormones on the progesterone-induced in vitro oocyte maturation and the non-hormonal induction of oocyte maturation, further studies of which can elucidate the mechanisms underlying the hormonal regulation of oogenesis in sturgeons. PMID- 12722590 TI - [Meiotic gynogenesis in the stellate, Russian sturgeon and sterlet]. AB - Diploid gynogenetic progenies were obtained in the stellate sturgeon Acipenser stellatus, Russian sturgeon A. gueldenstaedtii, and sterlet A. ruthenus by means of insemination of the eggs with UV-irradiated spermatozoa and suppression of the second meiotic division by heat shock. The gynogenetic nature of experimental fish was confirmed by RAPD-PCR analysis of DNA. Effective photoreactivation of UV induced lesions of spermatozoa was shown in the case of illumination of the fertilized eggs with visible light. This phenomenon should be taken into account when determining the doses of irradiation that allow inactivation of the male chromosomes and incubating gynogenetic embryos. Gynogenetic stellate and Russian sturgeons are viable and can be reared in order to study the mechanism of sex determination in sturgeons. PMID- 12722589 TI - [Protein kinase activities associated with actin cytoskeleton in the oocytes and eggs of African clawed frog]. AB - Protein phosphorylation with specific protein kinases plays the key role in the regulation of meiotic maturation of oocytes. However, little is known about the contribution of kinases to the temporal and positional regulation of the cytoskeleton rearrangement in maturing oocytes, including the actin cytoskeleton. In order to study a relationship between the kinase activities and actin cytoskeleton rearrangement, we analyzed protein phosphorylation in the isolated actin cytoskeleton of Xenopus laevis oocytes. Analysis of the full grown oocytes and eggs injected with [gamma-32P] "P has revealed phosphorylation of many proteins associated with the actin cytoskeleton and shown the appearance of three additional major phosphoproteins, 20, 43, and 69 kDa, during oocyte maturation. A significant number of these phosphoproteins were also found after incubation of the isolated cytoskeleton with [gamma-32P] "P in vitro, thus confirming that the kinases modifying these substrates are also specifically associated with actin. The in vivo and in vitro kinase activities were also stimulated during maturation. Analysis of kinase self-phosphorylation in situ and protein phosphorylation in solutions and substrate containing gels revealed a set of actin-associated kinases, including cAMP- and Ca(2+)-dependent kinases, as well as MAP, p34cdc2, and tyrosine kinase activities. Their level was the highest in the eggs. The involvement of kinases in the actin cytoskeleton rearrangement during oocyte maturation is discussed. PMID- 12722591 TI - [On the problem of formalization of temperature dependence of developmental rate in plants]. AB - Based on the data obtained in the experiments with constant and fluctuating diurnal temperatures, the temperature dependence of developmental rate was calculated for different photoperiodic groups of plants using the square equations of regression. These equations made it possible to calculate the temperature areas of maximum developmental rates of plants under different photoperiodic conditions. PMID- 12722592 TI - [Maturation of the follicle-enclosed common frog oocytes stimulated by low progesterone concentrations depends on transcription]. AB - The maturation of follicle-enclosed common frog oocytes stimulated by low progesterone concentration is inhibited by actinomycin D (5 micrograms/ml). The concentrations of progesterone, at which oocyte maturation was inhibited by actinomycin D, varied with the season and were different in different females. The inhibitor of steroidogenesis aminogluthetimide (100 micrograms/ml) did not suppress the maturation of follicle-enclosed oocytes in most experiments, which was induced by both high and low progesterone concentrations. The induction of maturation of the defolliculated ("denuded") oocytes required lower progesterone concentrations than of the follicle-enclosed oocytes and, in addition, the maturation of denuded oocytes was not suppressed by actinomycin D. Thus, it was shown for the first time that low doses of progesterone induced the maturation of amphibian oocytes while acting on the follicle wall cells and this process depended on transcription. The factor inducing or enhancing maturation, which is formed in the follicle cells in the presence of low progesterone concentrations, remains as yet unknown. PMID- 12722593 TI - [Effects of cycloheximide, aminoglutethimide, indomethacin, cytochalasin B and colchicine on ovulation and the ultrastructure of the ovarian follicle wall in the stellate sturgeon Acipenser stellatus Pall]. AB - The inhibitor of protein synthesis cycloheximide, inhibitor of steroidogenesis aminoglutethimide, and inhibitor of prostaglandin synthesis indomethacin, as well as the drugs affecting the cell cytoskeleton, such as cytochalasin B and colchicine, were used for studying the mechanisms of ovulation in the stellate sturgeon Acipenser stellatus Pall. Follicles were isolated from the body cavity within certain time intervals after the injection of pituitary suspension to a female and cultivated in media with the inhibitors. In the case of follicles isolated in the middle of the period from hormonal injection until ovulation, cycloheximide, cytochalasin B, and aminoglutethimide suppressed ovulation most effectively, while in the case of oocytes isolated during the last quarter of this period, aminoglutethimide and cytochalasin B were the most effective. It was shown using TEM and SEM that cycloheximide suppressed all processes related to the preparation for ovulation, except the initial ones: contraction of follicle cells and their processes and secondary flattening of these cells. In the presence of aminoglutethimide, the follicle cells underwent pathological changes. Incubation in the media containing indomethacin and colchicine prevented degradation of the outer theca layer at the follicle apex. In the presence of cytochalasin B affecting the cytoskeleton, the drawing of follicle cell processes from the jelly coat channels was blocked, the outer theca cells were strongly contracted, but the cell layer integrity was affected and it was divided in separate fragments. A relationship is discussed between the metabolic processes and morphological changes that lead to ovulation. It was proposed on the basis of the present and previous data that the preovulatory preparation of the follicle tissues comprises two contractile and two apoptotic processes distinctly coordinated in time and space. PMID- 12722594 TI - [The influence of thermoperiod on cucumber growth and development]. AB - We studied the influence of gradient temperature regimes on various parameters of the formation of above-the-ground and underground organs of cucumber plants, such as rate of leaf appearance, rate of growth, duration of growth and length of leaves, and the rate of growth of above-the-ground organs and roots. The plants were grown under the controlled conditions: at different combinations of day and night temperature, illumination 100 Wt/m2, and 12 h photoperiod. The comparison of constant and fluctuating diurnal temperature regimes has shown that in the optimal area for all studied indices, the highest values were recorded at the constant daily temperature (25 degrees C for all growth indices of above-the ground organs and 20 degrees C for growth of roots), while all gradient regimes either did not affect, or exerted inhibitory effects on the plant. The main acting fluctuating temperatures, that exerted stimulating effects, combined low hardening (15 degrees C) and optimal temperatures (25 degrees C), which was earlier described for animals. The 15/35 and 35/15 degrees C combinations were unambiguously inhibitory, since both temperatures are hardening for the cucumber. A lesser stimulating effect of the developmental rate in a plant, as compared to poikilothermic animals, could be due to a greater autonomy of plant ontogenesis because of autotrophy and, correspondingly, a greater degree of homeostasis. The mechanisms accounting for the reactions to temperature gradients are similar in different groups of ectotherms. PMID- 12722596 TI - Can you afford that capitation contract? Try this tool. PMID- 12722597 TI - Research suggests more care, greater expense fails to produce outcome improvements. PMID- 12722598 TI - New study quantifies impact of obesity on health care costs. PMID- 12722599 TI - IPA cap rates reflect California experience. PMID- 12722600 TI - Benchmark your medical management costs against this data set. PMID- 12722601 TI - [Dangerous recreational games]. PMID- 12722602 TI - [Feline immunodeficiency virus infection]. PMID- 12722604 TI - [Physiopathology of inflammation]. AB - The inflammatory reaction represents all the means associated with any aggression, and needed to maintain body integrity and survival. It acts first as a way to stimulate body surveillance and then as all the means to control any aggression. Such response largely non-specific of a specific cause, results from interactions between cells and soluble factors. The specificity of such response accounts for the anatomic localization of the disease. Among the many soluble factors, the proinflammatory cytokines tumour necrosis factor a and Interleukin-1 play a critical role, which has led to their therapeutic control with specific inhibitors. Positive clinical results have confirmed their biologic importance. However some side effects associated with the use of such inhibitors namely infections, have demonstrated the protective role of cytokines in stress survival. PMID- 12722603 TI - [Inflammation under the microscope]. AB - The diagnosis of an inflammatory tissular reaction is usual. The microscopical study gives a classification of the pattern of modification, of acute or chronic type, and teds to yield some clues in favor of the etiology. Routinely we perform a quantification of inflammation and fibrosis in some well defined areas of pathology (liver biopsies for example). PMID- 12722605 TI - [Acute-phase reaction]. AB - The systemic changes induced by inflammation have been referred as the acute phase response. The changes in the concentrations of acute-phase proteins are due largely to changes in their production by hepatocytes induced by pro-inflammatory cytokines. Because of its specificity, sensibility and short half-life, C reactive protein is the most useful indicator among all the acute-phase proteins. The clinical strategy to deal with an acute-phase response is to search the aetiology: infections, neoplasms, auto-immune and allergic diseases. The treatment of an acute-phase response is the treatment of its aetiology. PMID- 12722606 TI - [Corticosteroids and inflammation]. AB - Corticoids are molecules which the organism synthesizes to regulate a large number of immune and metabolic physiological mechanisms. The compounds used as therapeutic agents have at higher doses very useful anti-inflammatory and immunomodulatory properties. Corticoids have most original mechanisms of action which are essentially genomic (transcriptional) and characterized by activation (transactivation) or inhibition (transrepression) of numerous target genes. These molecules act in many cells involved in innate immunity (macrophages, granulocytes, mastocytes) and adaptive immunity (lymphocytes) but also in other cells (fibroblasts, epithelial and endothelial cells). The anti-inflammatory efficacy of corticoids may be explained by their inhibition of the synthesis of numerous cytokines, enzymes and mediators of inflammation or induction of cytokines and anti-inflammatory molecules (lipocortin). Corticoids also regulate cellular activation and survival processes (apoptosis), which explains their cytostatic efficacy in certain malignant hematological disorders. PMID- 12722607 TI - [Non-steroidal anti-inflammatory agents]. AB - Non steroidal anti-inflammatory drugs have the highest rate of prescription in the world due to their efficacy in counteracting inflammation, pain, fever and blood clot (with low-dose aspirin). This therapeutic class has been recently widen by a new family of NSAID, namely COX-2 selective inhibitors or coxibs. A 50%-reduction in the rate of gastric ulcers and bleeding has been achieved with coxibs. However, some questions remain unanswered yet, such as the cardiovascular issue with coxibs. Although the tolerability of NSAIDs is generally good, some well-established good clinical practices avoid the rare unexpected events which could sparsely occurred. PMID- 12722608 TI - [Anti-cytokines in the treatment of inflammation]. AB - Tumour necrosis factor (TNF) alpha and interleukine 1 (IL-1) have a pro inflammatory effect in all targets of the body. However, every inflammatory disease is characterized by a peculiar profile of secretion of cytokines. TNF alpha blocking agents are a major advance in the treatment of of rheumatoid arthritis, spondylarthropathies and Crohn disease. To date, the most common adverse events are reactions to infusions and reactivation of active tuberculosis with infliximab. Association of another immunosuppressive drug, such as methotrexate, decreases the first of these adverse events. IL1-RA is an inhibitor of IL-1 effects which have demonstrated its efficiency in rheumatoid arthritis. These anticytokines represent a dramatic progress in the treatment of inflammatory diseases but we have to stay very careful concerning the possible increased risk of infection or cancer. PMID- 12722609 TI - [Acute inflammation: when and how to treat?]. AB - Treatment of acute inflammation contains usually local or systemic anti inflammatory drugs. These medications, such as corticosteroids or NSAIDs, are not always safe to use. Major side effects are boosting secondary infections due to its immunosuppressive action or masking such as infections by its anti inflammatory activity. Because of these potential complications, the benefice/risk ratio of anti-inflammatory drugs must be weighed carefully besides its correct patterns of use and the prevention of its side effects. Available recommendations in the literature concerning its use are discussed thereafter. PMID- 12722610 TI - [What has become of inflammatory amyloidosis?]. AB - Inflammation associated amyloidosis (AA amyloid) remains a severe complication of chronic inflammatory diseases, namely of rheumatoid arthritis and of juvenile arthritis. Infectious causes of AA amyloid have become uncommon. In the course of chronic inflammatory diseases, amyloid should be regularly screened by proteinuria. The proof of amyloidosis requires tissue analysis examplifying specific apple-green birefringence after staining with Congo red. When amyloidosis is progressive in the course of one chronic disease, powerful anti inflammatory and immunosuppressive drugs can be used. PMID- 12722611 TI - [Problem-based learning: efficacy?]. PMID- 12722612 TI - [Obesity in the adult]. PMID- 12722613 TI - [Leg ulcer]. PMID- 12722614 TI - [Angina pectoris and myocardial infarction]. PMID- 12722615 TI - [Gastro-esophageal reflux in infants, children and adults. Hiatal hernia]. PMID- 12722616 TI - [Gait and balance disorders. Diagnostic approach]. PMID- 12722617 TI - [Urinary infections in children. Leukocyturia]. PMID- 12722618 TI - [New capacities of X-ray diagnosis in detection of small intestine pathology]. AB - The paper based on the data of more than 200 studies of the small bowel with the new agent Entero-VU specially designed for X-ray study of the bowel gives an objective evaluation of this agent. The authors of the paper analyze the capacities of Entero-VU in diagnosing 38 cases of Crohn's disease, tuberculosis and tumors of the small bowel. Moreover, as a separate section the paper provides an assessment of the agent in mapping the X-ray standards of the small bowel. In the authors' opinion, the use of this agent as though fills the existing gap in the X-ray study of the small bowel. PMID- 12722619 TI - [Results of point stenting of extended coronary stenosis in patients with ischemic heart disease]. AB - The paper analyzes the immediate and long-term results of point stenting in patients with extended (more than 30 mm) coronary stenoses and compares them with those obtained by routine stenting. The study included 177 patients with varying extent stenoses who underwent traditional implantation of intracoronary stents. Thirty-seven patients undergone the so-called point stenting formed a separate group. There was no significant difference in clinical and angiographic success rates between the patients of all three groups. Significant difference was also absent in subacute stent thrombosis between the patients of Groups 2 and 3 and those of the point stenting group. Thus, acute or subacute stent thrombosis was noted in 4 (2.3%) patients from the routine stenting groups, all the patients being from Groups 2 and 3, and in 2 (5.4%) from the point stenting group. No significant difference was found in the incidence of recurrent angina pectoris (restenosis) between Groups 1 and 2 and between Groups 2 and 3. At the same time, recurrent angina (restenosis) significantly more frequently developed in the patients of Group 3 than those of Group 1. As compared with Groups 2 and 3 patients, there was no significant difference in the incidence of recurrent angina (restenosis). PMID- 12722620 TI - [X-ray manifestations of severe congenital deformities of the visceral cranium]. AB - The paper discusses the most rational algorithm of an X-ray study and its results obtained while analyzing X-ray findings in 320 patients with severe congenital deformities of the visceral cranium. The malformations are shown to involve not only the visceral cranium, but also the cerebral cranium and its base and they can be rarely assigned to any of the known syndromes as they have frequently something in common. The most rational scheme of a X-ray study is presented. PMID- 12722621 TI - [Examining the effect of amiodarone on the lung: prospective follow-up data]. AB - Amiodarone (A) that belongs to Class III antiarrhythmics is a highly antiarrhythmic agents not only in treating, but in preventing various cardiac arrhythmias. However, its prolonged treatment may have an adverse life threatening effect on the lung in 1-10% of patients. The purpose of our study was to verify the safe effect of A on the lung. Forty-one patients receiving A were examined in the period of its saturation and for the following 6 months. A hundred and forty-nine patients treated with its maintaining doses long (for 11 years) were also studied. Lung X-ray films and spirograms were repeated every 1-3 months at the beginning of therapy and then every 6 months. Side pulmonary effects were not observed. The findings may lead to the conclusion that comparatively small maintaining doses of A are safe for the patient. PMID- 12722622 TI - [MRI in shoulder joint instability]. AB - The paper provides the results of MRI studies in 100 patients having complaints of pain and impaired movements in the shoulder joint in order to establish a diagnosis. Sixty-three patients were found to have MRI signs of shoulder joint instability (SJI). The paper presents and states the found MRI symptoms of SJI. The authors concludes that MRI of the shoulder joint in its instability should be used appropriately as it may early reveal changes in the articular osseous, cartilaginous, and soft tissues, which is useful in diagnosing and choosing a treatment. PMID- 12722623 TI - [Systems analysis of X-ray films of the vertebral column]. AB - The systems analysis of spinal X-ray films (SASXF) developed by the author solves the problem of an integral and, concurrently, detailed study of the spine of a patient on the basis of routine X-ray study. The examination of the patient gave rise to a patient's systems spinal model that considers the individual features of the vertebral column, which are most significant for a manual therapist's work. The standards of a X-ray study, a protocol form, and examination tools are described. An example of systems spinal model is given. The experience accumulated during studies of more than 2000 patients has shown that this procedure is highly effective in implementing medical manipulations by the methods of manual therapy. PMID- 12722624 TI - [Spiral computed tomography in the diagnosis of limb osteomyelitis]. AB - The results of radiation studies in 121 patients of different age (4 to 75 years) examined for limb osteomyelitis are analyzed. All the patients underwent routine X-ray study and computed tomography (CT), 26 patients had X-ray fistulography; 8, linear tomography; 10, CT fistulography; 6, scintigraphy, and 15, ultrasound study. Acute hematogenous osteomyelitis (AHO), chronic hematogenous osteomyelitis (CHO), and atypical (here Garre's sclerosing osteomyelitis and Brodie's abscess) osteomyelitis were ascertained in 10.6, 26.4, and 10.1% of cases, respectively. Posttraumatic osteomyelitis was diagnosed in almost 50% of the patients. CT defined the phase of chronic limb osteomyelitis. Spiral CT has proven to be the most effective technique for diagnosing limb osteomyelitis as compared with routine X-ray study: the accuracy of X-ray study was 81.8%, its sensitivity, 84.9%, and specificity, 60.0% and those of computed tomography were 96.7, 99.1, and 80.0%, respectively. PMID- 12722625 TI - [Bronchiolo-alveolar cancer: a case report]. PMID- 12722626 TI - [Diverticulum of the cardia]. PMID- 12722627 TI - [Intrathoracic and intraabdominal splenosis 40 years after splenectomy]. PMID- 12722628 TI - [Prospects of the use of non-ionic X-ray contrast media during endoscopic retrograde cholangiopancreatography]. PMID- 12722629 TI - [Genetic engineering of forest woody plants]. AB - The present state of genetic engineering (GE) of forest woody plants is considered with special reference to the materials of the International Conference "Wood, Breeding, Biotechnology and Industrial Expectations" held in France in June, 2001. Main tree species subjected to GE are listed, aims of constructing transgenic plants discussed, and methods described. Major achievements in the field are considered along with the problems associated with the employment of GE in the breeding of forest woody plants. PMID- 12722630 TI - [Cloning and expression of the Mycoplasma hominis ftsZ for a cell division protein]. AB - A Mycoplasma hominis chromosomal fragment containing the full-length ftsZ gene was cloned and sequenced. Natural expression of this gene was demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) with total RNA. The M. hominis FtsZ protein was shown to differ substantially from its counterparts of two other Mycoplasma species, M. genitalium and M. pneumoniae. The possibility of M. hominis ftsZ expression in Escherichia coli was demonstrated with several bacterial strains. The M. hominis FtsZ protein was isolated from E. coli cells transformed with recombinant plasmids carrying the M. hominis ftsZ gene. Complementation between the E. coli and M. hominis FtsZ proteins was observed in transformants. PMID- 12722631 TI - [Determination of functional role of nucleotide composition in the transcription start region of the Escherichia coli udp gene]. AB - The promoter of the Escherichia coli udp gene contains the poly-T (5'-TTTTT-3') motif in the transcription start region located at the distance of 3 nucleotides with respect to the Pribnow box. By means of site-directed mutagenesis, mutations in the +1, -1, and +3 positions of this region were isolated and their functional role in transcription initiation was determined. It was shown that in addition to the thymine nucleotide earlier identified at position 4 (with respect to the 5' end of the poly-T motif), the third thymine nucleotide may serve as an efficient transcription site. The functional significance of the presence of the poly-T motif in the transcription initiation region is discussed. Analysis of the isolated mutant promoters revealed the ability of the guanine nucleotide at the +3 position to stabilize the formation of a productive transcription complex and to serve as an additional transcription start. PMID- 12722632 TI - [The prqA and mvrA genes encoding carrier proteins control resistance to methyl viologen in the cyanobacterium Synechocystis sp. PCC6803]. AB - Derivatives with insertional inactivation of prqA and mvrA genes were obtained and studied in the Synechocystis sp. PCC6803 wild-type strain and in the mutant Prq20 resistant to methyl viologen (MV). It was shown that the formation of resistance to MV is associated with the operation of two systems: constitutive and inducible. The prqA gene encoding drug efflux proteins controls the constitutive system of cell resistance to MV. Derepression of the prqA gene is the main reason for an enhanced MV resistance in the Prq20 mutant with impaired repressor function of the PrqR protein. The mvrA gene encoding the transmembrane protein from the family of transporters of sugar and other compounds controls the inducible MV resistance. It is assumed that the MvrA protein is required for efficient elimination from cells of toxic substances formed upon oxidative stress or participates in the repair of membranes destroyed by oxidants. The data obtained demonstrated for the first time that transport systems are involved in the formation of MV resistance in photosynthetic organisms. PMID- 12722634 TI - [Comparative analysis of MGE 412 patterns in 18 isogenic lines of Drosophila melanogaster]. AB - Comparative analysis of patterns of mobile genetic element 412 was conducted in 18 isogenic lines of Drosophila melanogaster isolated in three isogenic experiments in 1987 through 1999. Twelve "extra-hot" isogenization sites (in 15 18 lines) and 23 "hot" isogenization sites (> or = 10 lines) were found; of these, 19 occurred in the original heterogeneous line. These sites virtually do not overlap with hot induction sites of transposition. Sites of the latter group generally retain their positions during isogenization. It was shown that no more than 20% of the new sites were brought from the balancer by double recombination, while inbreeding and outbreeding caused 80% of them. Different factors were shown to have different hot isogenization sites. A similarity tree was constructed for the patterns of 18 isogenic lines. The maximum peak of the tree was very low (< 0.25), i.e., the isogenic lines are more similar to than different from one another. The tree was subdivided into subtrees. The division was in good agreement with the isogenization groups corresponding to individual isogenization experiments. Significant correlation was found between the total fragment length and the number of new sites per lines. PMID- 12722633 TI - [Semidominant effect of the l(1)ts403 (sbr10) mutation on nondisjunction of sex chromosomes in meiosis in Drosophila melanogaster females exposed to heat]. AB - The sbr gene of Drosophila melanogaster belongs to the NXF (nuclear export factor) family responsible for the mRNA transport from nucleus to cytoplasm. We have shown that in the heat-exposed (37 degrees C, 1 h) females, the l(1)ts403 (sbr10) mutation leads, in particular, to the high-frequency nondisjunction and loss of sex chromosomes in meiosis. For this trait, the incomplete dominance of the sbr10 mutation is observed. At the same time, the sbr10 mutation is recessive for many other traits of the heat-exposed flies: reduced viability, low fertility, impaired synthesis of the heat shock proteins, etc. The females heterozygous for the null allele (Df(1)vL4, a deletion eliminating gene srb) do not differ from females homozygous for the wild-type allele in frequency of the heat shock-induced nondisjunction and loss of sex chromosomes in meiosis. Because of this, the sbr10 mutation can be assigned to the gain-of-function alleles (those gaining the dominance function). Expression of the mutant sbr10 allele against the background of the wild-type allele suggests that in the heat shock exposed females, the heat-modified product of this ts allele has a strong effect on sex chromosome disjunction in meiosis. PMID- 12722635 TI - [A method for preparation of synaptonemal complexes of meiotic chromosomes from basidial protoplasts of the common mushroom Agaricus bisporus (Lange) Imbach]. AB - For the first time, preparations of synaptonemal complexes (SCs) were made from meiotic chromosomes of white button mushroom (Agaricus bisporus) basidia. It is the first experience of obtaining SC preparations of filamentous fungi from isolated meiosporangium protoplasts. Previously, only yeast SC preparations were obtained following this approach. The method includes four major stages: isolation of basidium protoplasts by treatment of basidia with lytic enzymes, spreading of protoplast nuclei on a filmy support by osmotic shock, staining the preparations with silver nitrate, and examination under light and electron microscopes. The structures of spread premeiotic nuclei, axial elements of chromosomes, SCs, chromatin, and nucleoli were studied at the leptotene-diplotene stage of meiotic prophase I. PMID- 12722637 TI - [The effect of genotypes of parental rye lines on the development of quantitative traits in primary octoploid Triticale: spike fertility]. AB - The number of seeds and seed setting in the main spike were studied in primary octoploid triticale obtained from crosses between the common wheat cultivar Chinese Spring and 66 inbred rye lines. In some rye lines, the mutations of self fertility were identified in the S, Z, or T incompatibility loci. The number of seeds was determined under controlled self-pollination of the main spike. In the set of triticale examined, each trait exhibited high variation. Hence, the rye lines were suggested to carry gene alleles both increasing and decreasing these traits in triticale. All the traits studied were significantly influenced by environmental conditions. Ten triticale lines were identified, which had the largest seed setting under self-pollination. Seven out of ten samples with the high number of seeds carried mutations in the T locus and in the three samples, the unidentified self-fertility mutations were present. The triticale lines with mutations in the S and Z loci displayed much lower self-fertility on average. The ways and means of identifying and mapping the rye gene responsible for distinctions between the triticale quantitative traits are discussed. PMID- 12722636 TI - [Abnormal condensation of meiotic chromosomes caused by the mei8 mutation in rye Secale cereale L]. AB - Inheritance of two spontaneous meiosis-specific mutations with similar cytologic phenotype was studied. Both mutations were independently obtained from two rye populations (Vyatka variety and weedy rye). Both mutations are recessive, allelic, and monogenically inherited; the corresponding gene is designated mei8. The mutant alleles of the gene cause abnormal meiotic chromosome structure expressed as irregular compaction along the chromosome length, chromatin stickiness at all stages of meiosis, and chromosome fragmentation in anaphase I. PMID- 12722639 TI - [Genetic mechanisms of resistance and susceptibility to leukemia in Ayrshire and black pied cattle breeds determined by allelic distribution of gene Bola-DRB3]. AB - In the herds of Ayrshire and Black Pied cattle breeds of Russian selection, comparative analysis of allelic distribution of BoLA-DRB3 was performed in animal groups with different status of persistent lymphocytosis (PL) caused by the bovine leukemia virus (BLV). Alleles were typed by PCR-RFLP. Different spectra of BoLA-DRB3 alleles mediating susceptibility and resistance to leukemia were detected in the studied breeds. The role of amino acid motives in beta 1 domain of BoLA-DRB3 antigens was confirmed: ER (in positions 70-71), in resistance to leukemia and VDTY and VDTV (75-78), in susceptibility to leukemia. The nucleotide sequence of allele BoLA-DRB3.2*7 with deletion of codon 65, which resulted in the changed conformation of the corresponding antigen molecule, was associated with resistance to PL. Cows of Black Pied and Ayrshire breeds with genotypes coding VDTY/VDTV (RR = 11.67, P = 0.014) and VDTY/VDTY (RR = 4.71, P = 0.022), respectively, were shown to be susceptible to PL. The role of heterozygosity level was demonstrated (estimated by BoLA-DRB3 alleles and by amino acid motives in positions 75-78 of the antigen) as an unspecific factor of resistance to PL. The lowest heterozygosity level by amino acid motives (75-78) was revealed in PL animals, for which sample inbreeding coefficients were detected: F = 0.324 and F = 0.084 in Ayrshire and Black Pied breeds, respectively. PMID- 12722640 TI - [Interaction of the diabetes insipidus locus alleles with the renal 120 kDa protein-encoding gene in rat development]. AB - Age-dependant dynamics of the kidney inner medullary 120-kDa protein content in vasopressin-deficient Brattleboro rats with di/di mutant genotype was studied in comparison with WAG rats with genotype and normal vasopressin expression. Age dependant dynamics of vasopressin content in neurohypophysis of WAG rats was also examined. It was shown that 10-day-old WAG rats were unable to elevate the synthesis of the 120 kDa protein in respond to long-term dehydration, while this tendency was clearly observed in the 15-day-old rats and later in the development. In WAG rats the onset of this specific feature was time correlated with the development of the respond to hydration by the elevation of vasopressin synthesis and release from neurohypophysis into blood. In the di/di rats dehydration had no effect on the kidney 120-kDa protein synthesis in all ages examined. These results point to the interaction between the di alleles and the 120-kDa protein-encoding gene in the course of development. PMID- 12722638 TI - [Expression of a thermostable bacterial cellulase in transgenic tobacco plants]. AB - The bacterial gene of the thermostable endo-beta-1,4-glucanase (cellulase) was shown to retain its activity and substrate specificity when expressed in transgenic tobacco plants. The leader peptide of the carrot extensin was efficient in transferring the bacterial enzyme into the apoplast. The expression of the bacterial cellulase gene leads to changes in the plant tissue morphology. In the transgenic plant lines, regeneration of primary shoots from callus occurred at the three to five times higher cytokinin (6-BAP) concentration than in control plants. The transgenic plants that expressed the bacterial gene exhibited increased business and altered leaf shape. The transgenic plants developed can be used as models for studying the cellulases role and function in plants. PMID- 12722641 TI - [Genetic changes in pink salmon Oncorhynchus gorbuscha Walbaum during acclimatization in the White sea basin]. AB - Genetic parameters of pink salmon introduced into the White Sea basin in 1985 and 1998 were compared to the corresponding parameters of the donor population from the Ola River (Magadan oblast). The detected genetic differences indicate that colonization of a new area is accompanied by impoverishment of the gene pool of the native population. This effect was particularly marked in the odd-year line of pink salmon introduced in 1985. The probable causes of these genetic changes are discussed. PMID- 12722643 TI - [Chromosomal studies of male infertility]. AB - Prometaphase and metaphase chromosome analyses performed on 70 consecutive men with primary infertility (for a period of at least 2 years) revealed 8 (11.42%) men with some kind of chromosomal abnormality. The highest frequency of abnormal karyotypes (10%) was found among patients with azpospennia and the most frequent anomaly was 47, XXY chromosomal constitution, found in 6 (8.57%) patients. All the chromosomal aberrations found in this study was sex chromosomal type and we did not find any autosomal aberration. All patients with numerical chromosomal anomalies had azoospermia. The incidence of structural aberration in our study was 1.42%. Fifteen patients had different chromosomal variants (21.38%). We suggest that men with azoospermia should be considered for cytogenetic investigation and we report that "variants of the Y chromosome" have no influence on the sperm count (million/ml) and fertility of men. PMID- 12722642 TI - [Genetic and epidemiological studies in Dagestan highland isolates]. AB - Interpopulation differences in the epidemiology and age of onset of complex diseases, as well as expression of some vital parameters, have been found. The relationship between these interpopulation differences and the genetic processes that have been occurring in the populations throughout their history has been demonstrated. The Daghestan genetic isolates studied are characterized by aggregation of certain complex diseases. In each genetic isolate, almost all affected subjects with homogeneous clinical phenotypes belong to the same large pedigree with a limited number of founders. There is evidence for a large variance of the population risk of schizophrenia (morbid risk) in Daghestan isolates (this parameter varies from 0 to 5%). Examination of 211 cases of schizophrenia earlier diagnosed in Daghestan psychiatric hospitals has shown that only 139 of them meet the DSM-IV criteria for schizophrenia. The remaining 72 subjects have, according to DSM-IV criteria, various schizoaffective and affective disorders; all of these subjects are close relatives of the schizophrenic patients. The age of onset of schizophrenia in the isolates studied varies from 14 to 40 years (20.84 +/- 0.568 years). Offspring of consanguineous marriages exhibit later age at onset and a higher risk of schizophrenia than offspring of exogamous marriages. The results of multivariate genetic analysis indicate that different gene complexes are involved in the pathogeneses of early onset and late-onset forms of schizophrenia. An association of schizophrenia incidence, its age dependence, and reproductive parameters with polymorphisms of some microsatellite loci have been demonstrated. PMID- 12722644 TI - [The frequency of heteroploidy in spermatozoa of patients with infertility]. AB - The incidence of heteroploid spermatozoa was studied by mono- and dual-colored fluorescence in situ hybridization in semen samples from donors and patients with normal and impaired spermatogenesis. The frequency of heteroploid sperm in the ejaculate was linearly and inversely correlated with sperm parameters (sperm concentration in the ejaculate and proportion of motile and morphologically normal spermatozoa). The level of heterploidy was the most significant in the semen samples from patients with oligoasthenoteratospermia and oligoasthenospermia. PMID- 12722646 TI - [Molecular genetic diagnostics of some Daphnia species (Crustacea, Cladocera) from the Volga river]. AB - Population genetic structure of the widespread Daphnia species from Kuibyshev and Saratov reservoirs was examined by use of RAPD-PCR technique with four mitochondrial DNA (16S rRNA) specific primers. One of the examined Daphnia clones from the Volga region appeared to be conspecific to the North American Daphnia galeata, while another clone was most likely a hybrid between D. galeata and D. cucullata. PMID- 12722645 TI - [Analysis of the allelic polymorphism of the five X-linked (CA)n dinucleotide repeats in Russia]. AB - A PCR-based survey of allelic polymorphism of three microsatellite markers, DXS998, DXS548, and FRAXAC1, mapped to chromosome region Xp27.3, and two microsatellite markers, DXS8091 and DXS1691 located on Xq28 was carried out using a series of DNA samples obtained from 98 unrelated individuals from Russia. The number of alleles detected on electrophregrams for each marker tested was 4, 6, 4, 5, and 3, respectively. The values of heterozygosity index for the markers examined were 0.65, 0.27, 0.38, 0.70, and 0.29, respectively. The observed distribution of the allelic frequencies for each microsatellite marker examined fitted Hardy--Weinberg expectations. The values of individualization potential determined for each marker were 0.24, 0.53, 0.43, 0.12, and 0.52, respectively. In the sample tested the genotype distribution with regard to above loci was determined. The perspectives of using the analyzed allelic polymorphisms for indirect DNA diagnostics of the monogenic diseases located in this chromosome region (X-linked mental retardations, FRAXA and FRAXE) as well as for human population genetics and personal identification is discussed. PMID- 12722647 TI - [Multiple functions of corrinoids in prokaryote biology]. AB - Data on more than 30 metabolic processes and biochemical reactions, involving corrinoids, which have been described in prokaryotes thus far, are reviewed. These pathways (central or specific, catabolic or anabolic) are inherent in bacteria and archebacteria of diverse phylogenetic lineages, comprising several physiological groups. Particular emphasis is placed on the role of corrinoid dependent transmethylation in acetogenesis and methanogenesis and on the contribution of adenosylcobalamin in DNA metabolism. PMID- 12722648 TI - [Kinetics of catalase inactivation induced by ultrasonic cavitation]. AB - Kinetic patterns of sonication-induced inactivation of bovine liver catalase (CAT) were studied in buffer solutions (pH 4-11) within the temperature range from 36 to 55 degrees C. Solutions of CAT were exposed to low-frequency (20.8 kHz) ultrasound (specific power, 48-62 W/cm2). The kinetics of CAT inactivation was characterized by effective first-order rate constants (s-1) of total inactivation (kin), thermal inactivation (*kin), and ultrasonic inactivation (kin(us)). In all cases, the following inequality was valid: kin > *kin. The value of kin(us) increased with the ultrasound power (range, 48-62 W/cm2) and exhibited a strong dependence on pH of the medium. On increasing the initial concentration of CAT (0.4-4.0 nM), kin(us) decreased. The three rate constants were minimum within the range of pH 6.5-8; their values increased considerably at pH < 6 and pH > 9. At 36-55 degrees C, temperature dependence of kin(us) was characterized by an activation energy (Eact) of 19.7 kcal/mol, whereas the value of Eact for CAT thermoinactivation was equal to 44.2 kcal/mol. Bovine serum and human serum albumins (BSA and HSA, respectively) inhibited sonication-induced CAT inactivation; complete prevention was observed at concentrations above 2.5 micrograms/ml. Dimethyl formamide (DMFA), a scavenger of hydroxyl radicals (HO.), prevented sonication-induced CAT inactivation at 10% (kin and *kin increased with the content of DMFA at concentrations in excess of 3%). The results obtained indicate that free radicals generated in the field of ultrasonic cavitation play a decisive role in the inactivation of CAT, which takes place when its solutions are exposed to low-frequency ultrasound. However, the efficiency of CAT inactivation by the radicals is determined by (1) the degree of association between the enzyme molecules in the reaction medium and (2) the composition thereof. PMID- 12722649 TI - [Induction of endo-1,4-beta-xylanase and beta-galactosidase in the original and recombinant strains of the fungus Penicillium canescens]. AB - The induction of the synthesis of secreted enzymes endo-1,4-beta-xylanase (EC 3.2.1.8) and beta-galactosidase (EC 3.2.1.23) in the original and recombinant Penicillium canescens strains has been studied. In all producer strains, the synthesis of these enzymes was induced by arabinose and its metabolite arabitol. The two enzymes differed in the concentration of arabinose required for the induction: the synthesis of beta-galactosidase was most pronounced at 1 mM, whereas maximum synthesis of endo-1,4-beta-xylanase was observed at 5 to 10 mM. An increase in the number of endo-1,4-beta-xylanase copies in the high-copy number strain of the fungus suppressed the synthesis of beta-galactosidase; the synthesis of endo-1,4-beta-xylanase in the high-copy-number recombinant producing beta-galactosidase was affected to a lesser extent. The amount of the enzymes synthesized did not depend on the saccharide used as a sole source of carbon for growing the mycelium prior to its transfer to the inducer-containing medium. PMID- 12722650 TI - [Identification of a new steroid transforming strain of mycobacteria as Mycobacterium neoaurum]. AB - The ability to utilize sterols as a sole source of carbon was studied in 80 strains and consortia of hydrocarbon-oxidizing bacteria. One of the strains, which efficiently transformed both individual sterols and their mixtures, was identified as Mycobacterium neoaurum, based on the analysis of the sequence of the 16S rRNA gene. PMID- 12722651 TI - [Regulation of acetate metabolism in a strain of Acinetobacter sp., growing on ethanol]. AB - Ethanol metabolism in Acinetobacter sp. is limited by the rate of acetate assimilation in a reaction catalyzed by acetyl-CoA synthetase (EC 6.2.1.1). Effects of ions (sodium, potassium, and magnesium), byproducts of ethanol and acetaldehyde oxidation (NADH and NADPH), and pantothenic acid on this enzyme have been studied (sodium, NADH, and NADPH inhibit acetyl-CoA synthetase; pantothenic acid, potassium, and magnesium act as the enzyme activators). Conditions of culturing were developed, under which ethanol, acetaldehyde, and acetate in Acinetobacter cells were oxidized at the same rates, producing a threefold increase in the activity of acetyl-CoA synthetase in the cell-free extract. The results of studies of acetyl-CoA synthetase regulation in a mutant strain of Acinetobacter sp., which is incapable of forming exopolysaccharides, provide a basis for refining the technology of ethapolan production, involving the use of C2 substrates. PMID- 12722652 TI - [Kinetic parameters of a culture of the hydrogen-oxidizing Ralstonia eutropha, grown under the regimen of biosynthesis of polyhydroxybutyrate]. AB - Kinetic parameters of a culture of the hydrogen-oxidizing bacterium Ralstonia eutropha, grown on a gas substrate under the conditions favoring autotrophic biosynthesis of polyhydroxybutyrate, were studied. The following parameters, making it possible to control and optimize the process in industrial situations, were determined: specific rate of substrate consumption, physical properties of culture medium, and coefficients of heat emission and mass transfer. PMID- 12722653 TI - [Effect of toxic concentrations of 2,4,6-trinirtotoluene on the physical properties and morphology of bacillus subtilis SK1]. AB - It has been demonstrated for the first time that the toxic effect of 2,4,6 trinitrotoluene on the gram-positive strain Bacillus subtilis SK1 is accompanied by a decrease in the cell size and an increase in the refraction index (i.e., density) and thermostability of the culture. These data suggest that determination of kinetic parameters of bacteria growing under the conditions of toxic stress from optical density measurements may result in erroneous conclusions. PMID- 12722654 TI - [Reversed-phase high performance liquid chromatography of the products of microbiological degradation of toluene ]. AB - Conditions have been selected for a reversed-phase high-performance liquid chromatographic assay of intermediate products formed in the course of utilization of toluene by Pseudomonas putida. The composition of products indicates that degradation of toluene by strain BS590-P proceeds primarily through the formation of benzoate and catechol. This is followed by degradation of catechol via ortho-cleavage. In strain BS3701-P, toluene oxidation involves both the side chain and the aromatic ring. PMID- 12722655 TI - [Extracellular protein metabolite of Luteococcus japonicus subsp. casei reactivates cells subjected to oxidative stress]. AB - A protein exometabolite isolated from the culture liquid of Luteococcus japonicus subsp. casei reactivates the cells of this microorganism, following H2O2 or paraquat-induced oxidative stress. The resistance of L. casei cells to these oxidizers is accounted for by the high activity of superoxide dismutase and catalase. The effect of the protein exometabolite is universal, in that it reactivates the cells after UV irradiation, heating, or oxidative stress. However, the cells subjected to oxidative stress are significantly less susceptible to the reactivating effect, as compared to their UV-irradiated or heated counterparts. Possible causes of these differences are discussed. PMID- 12722656 TI - [Dependence of the composition of the avermectin complex of Streptomyces avermitilis on glucose content of the growth medium]. AB - The effects of glucose concentration in the medium and O-methyl-L-threonine resistance on the ratio of components of the avermectin complex produced by Streptomyces avermitilis have been studied. Glucose deficiency increases the ratio of components A and a, while decreasing that of components 1 in the complex. A mutation that renders the microorganisms resistant to O-methyl-L threonine (an analog of isoleucine) increases the ratio of components a, while decreasing that of components 1 in the complex. The distribution of a and b in fractions 1 and 2 remains constant: the values of the ratio a/b in the fractions amount, respectively, to 1:1 and 2:1. The relation of the variations in the composition of the avermectin complex to changes in the carbohydrate metabolism of the producer stain, underlain by availability of the source of carbon, is discussed. PMID- 12722657 TI - [Effect of various sources of nitrogen and carbon on the biosynthesis of proteolytic enzymes in a culture of Aspergillus awamori 21/96]. AB - Effects of various conditions of nitrogen and carbon nutrition on the biosynthesis of proteolytic enzymes in a selected culture of Aspergillus awamori 21/96 were studied. This strain was shown to produce proteolytic enzymes constitutively. In the presence of mineral sources of nitrogen, the synthesis of the enzymes under study was not induced by proteinaceous substrates. Optimum conditions of the enzyme biosynthesis were achieved with casein as a source of nitrogen and starch or dulcitol as a source of carbon (which increased the production of the enzymes 1.7 and 8 times, respectively). When the cells were grown on starch, their specific activity exceeded control levels 18 times. PMID- 12722658 TI - [Occurrence of indole alkaloids among secondary metabolites of soil Aspergillus]. AB - The occurrence of indole alkaloids among secondary fungal metabolites was studied in species of the genus Aspergillus, isolated from soils that were sampled in various regions of Russia (a total of 102 isolates of the species A. niger, A. phoenicis, A. fumigatus, A. flavus, A. versicolor, A. ustus, A. clavatus, and A. ochraceus). Clavine alkaloids were represented by fumigaclavine, which was formed by A. fumigatus. alpha-Cyclopiazonic acid was formed by isolates of A. fumigatus, A. flavus, A. versicolor, A. phoenicis, and A. clavatus. The occurrence of indole containing diketopiperazine alkaloids was documented for isolates of A. flavus, A. fumigatus, A. clavatus, and A. ochraceus. No indole-containing metabolites were found among the metabolites of A. ustus or A. niger. PMID- 12722659 TI - [Production of fumonisins by Fusarium moniliforme strains isolated from corn grain]. AB - Fusarium moniliforme is the predominant fusarium species in the grain mycoflora of corn grown in Northern Caucasus, accounting for 95% of fusarium isolates. Eighty-five Fusarium moniliforme strains were grown on grain substrate and checked for the presence of fumonisins (B1 + B2 + B3) by indirect solid-phase enzyme immunoassay (EIA). All strains were capable of producing fumonisins (0.95 to 32,000 mg/kg). Strains sampled in the Krasnodar krai produced the highest fumonisin levels (averaging 5490 mg/kg). PMID- 12722660 TI - [Products of spontaneous conjugation of aflatoxins with bovine serum albumin: immunochemical properties]. AB - Products of spontaneous conjugation of aflatoxins B1, G1, and G2 with bovine serum albumin (BSA) were shown to interact with antibodies against aflatoxins. Solid-phase BSA conjugates inhibited the binding of aflatoxins by anti-aflatoxin antibodies. Antisera against BSA-B1, BSA-G1, and BSA-G2 were obtained and their specificity, determined. The mechanisms of spontaneous binding of aflatoxins by proteins are discussed. PMID- 12722661 TI - [Effect of ambiol and 2-chloroethylphosphonic acid on the content of phytohormones in potato leaves and tubers]. AB - The effects of the antioxidant Ambiol and 2-chlorethylphosphonic acid (2-CEPA) on individual concentrations and concentration ratios of phytohormones, photosynthesis and photophosphorylation rates, sucrose and starch content in tubers, and plant productivity were studied in potato (Solanum tuberosum L). Ambiol increased the ratio of indoleacetic acid (IAA) to abscisic acid (ABA), IAA/ABA, and that of zeatin (Z) and zeatin riboside (ZR) to ABA, (Z + ZR)/ABA. These effects were underlain by an increase in the content of auxins and cytokinins and a decrease in ABA. Unlike Ambiol, 2-CEPA increased the level of ABA, the effect being the most pronounced in the tubers. Ambiol increased the rates of photosynthesis and noncyclic photophosphorylation in chloroplasts isolated from potato leaves. The relation of this phenomenon to auxin and cytokinin accumulation, Ambiol- and 2-CEPA-induced changes in the hormonal balance of potato tubers, carbon metabolism, and plant productivity is discussed. PMID- 12722663 TI - [New method for detection of enzymatic activity of flavocytochrome b2 in electrophoregrams]. AB - A new method of visualization of the activity of flavocytochrome b2 (FC b2; L lactate: ferricytochrome c oxidoreductase, EC 1.1.2.3) in electrophoretograms was developed, based on the interaction between ferrocyanide (generated during the enzymatic reaction) and Fe23+, resulting in the formation of intensely colored precipitates of Berlin blue. The main advantages of this method were its high sensitivity (less than 0.005 U FC b2 was detected within a suitable time period) and the stability of the dye formed. The method developed can be used for determining FC b2 activity in cell-free extracts (e.g., in the selection of FC b2 producers) and monitoring chromatographic purification of proteins, as well as in other cases associated with FC b2 assessment. PMID- 12722662 TI - [Binding of aliphatic aldehydes by cornstarch polysaccharides]. AB - Binding by cryotextured cornstarch of individual aliphatic aldehydes (C6-C10; saturated or unsaturated) and their mixtures from aqueous solutions has been studied using capillary gas chromatography. The amount of compounds absorbed by the cryotextures depended linearly on the concentration of aldehydes in the original gel. The majority of the compounds under study were bound irreversibly. Aldehydes with low molecular weight were better absorbed by the cryotextures than by granules of intact cornstarch. Data of IR spectrocopy demonstrated that binding to cornstarch polysaccharides decreased the conformational mobility of odorants. The appearance of binding isotherms depended of the extent of aldehyde sorption, suggesting the involvement of multiple mechanisms of binding. The formation of supramolecular complexes through cooperative hydrophobic interactions between aldehydes and cornstarch polysaccharides was the preferential mechanism of the sorption. PMID- 12722664 TI - [Surgical education in medical school]. AB - Progress of medicine and technology is quick. Accumulation of knowledge is immense and advancement of surgical technologies is vast. Curriculum and methods of surgical education in medical school should be modified accordingly. Our society has not actively participated in surgical education in medical school but considering the importance of surgical education in medical school for the future of surgery and surgical practice, and society (JSS) should more actively participate in surgical education for medical students by starting a special committee for this particular purpose within the society. A desirable curriculum in surgical education in medical school is presented. PMID- 12722665 TI - [Compulsory postgraduate clinical training and surgical education]. AB - In April 2004, a new training system for physicians and surgeons will take effect in Japan. All medical graduates, after passing the National Medical Practitioners Examination, are to complete a two-year compulsory clinical training program, which includes rotations to the clinical services of internal medicine, surgery, emergency medicine, pediatrics, obstetrics and gynecology, psychiatry, and regional primary practice. Hospitals offering training programs must be approved by the Ministry of Health, Welfare and Labor. The trainees will be provided with reasonable stipends. In accordance with the introduction of new system, the current surgical training system is to be renovated and the terms for surgical trainees to be eligible for surgical specialty boards must be reviewed. PMID- 12722667 TI - [Postgraduate trauma education for surgeon--from the veiwpoint of trauma and emergency physicians]. AB - A recent important topic in the field of trauma care is the elimination of "preventable death." Since patients' satisfactory prognosis and good recovery mainly depend upon the success of the initial evaluation and care, the responsibility of physicians who first come into contact with trauma patients in the emergency room is especially heavy. Many general surgeons examine trauma patients, although, unfortunately they have few opportunities for training in how to deal with those patients in the present educational program. Traumatology should be one of the most important areas of surgery, and therefore this paper emphasizes that every surgeon needs to be educated and trained in an adequate curriculum of trauma care. The Japanese Association for Acute Medicine could help by offering the Japan Advanced Trauma Evaluation and Care (JATEC) training program. JATEC proposes standardized guidelines as well as a nationwide the educational program for trauma management. PMID- 12722666 TI - [The new Japanese board of surgery for general surgery specialists]. AB - The postgraduate training and board certification system for general surgery has just reached a new era with the upgrading from the previous system for registered surgeons. The new system consists of first-step training of a minimum of 5 years (initially 4 years or more) covering general surgery, emergency medicine, and basic knowledge of subspecialties. The subspecialties programs are optional but combined as the second step, and 7 years of training including the first step are required. The new system of a general surgical training consists of a residency training program with registered hospitals and a minimum requirement of surgical experience. At this starting point, the Japan Surgical Society should take responsibility for this new training system, together with the related societies of subspecialties, in this new step for the official and adequate public qualification of surgeons. PMID- 12722668 TI - [Specialist of general surgery and gastroenterological surgery]. AB - The educational committee of the society of gastroenterological surgery made a rule that the specialist of gastroenterological surgery must have a title of a general surgeon which was approved by the educational committee of the Japan surgical society. This means that a specialist of gastroenterological surgery should be able to treat diseases in the area of the general surgery and should be able to do the primary cares for whole areas of gastroenterological diseases which are the upper and the lower intestinal diseases, hepato-bilial and pancreatic diseases in addition. The Japan society of gastroenterological surgery offers a 7 years long gastroenterological surgery training curriculum after graduate from a medical school to train the specialist of Gastroenterological surgery. In this 7 years training period, the Japan surgical society also offers a 5 years long general surgery training course in the first 5 years. These two courses are proceeding in parallel. The essential contents of these two courses are overlapped, so these common contents may count as experiences for both courses respectively. PMID- 12722669 TI - [Surgical education and surgical board system from the view point of pediatric surgeons]. AB - It is necessary to give priority to training in problem-solving ability regarding pre- and postoperative care during undergraduate education and internship training. We do not need to give priority to instruction in the knowledge of a specialized surgical field before surgical residency. Moreover, education that emphasizes a subspecialty from the early stage of surgical residency does not answer the requirements of society, while training in general surgery is indispensable. Reevaluation from the viewpoints of a number of specialists and regional distribution is requested for the present board system, at least in pediatric surgery. In addition, major issues like the acquisition of corporate status of a related surgical society with specialists' advertising deregulation and the influence of the introduction of required two-year clinical training should be resolved. PMID- 12722670 TI - [Training of cardiovascular surgery]. AB - Among all the other fields in medicine where safety and good quality are always expected, cardiovascular surgery is the one field where you are expected the most. Therefore cardiovascular surgeons should have an ability to deal adequately with a severe relationship between patients and surgeons. A long-term education after several years of basic surgery training is mandatory to be an authorized high quality cardiovascular surgeon. However, basic surgery is a synonym of general surgery in the present system. In order to authorize general surgeons as well, basic surgery and general surgery should be separated. To avoid the world of cardiovascular surgery becoming the aging society, a long-term training of cardiovascular surgery should begin as early as possible after efficient short term training of basic surgery. Meanwhile high income should be guaranteed for high risk/volume work. PMID- 12722671 TI - [Medical education and surgical residency program for general thoracic surgery]. AB - We introduced medical education for general thoracic surgery to medical students and a new surgical residency program to instruct residents who aspire to become general thoracic surgeons. It is important to understand the etiology of thoracic diseases, methods of diagnosis for tumorous lesions, anatomy of the interior thorax, surgical therapies and indications for thoracic diseases, and management of perioperative periods in the medical education of general thoracic surgery. According to the new residency program, a surgical resident must complete a super rotating system that consists of the subjects of internal medicine, general surgery, emergency and critical care medicine, anesthesiology, pediatrics, gynecology, and local healthcare for several months each in the first two years. On the other hand, a surgical residency program includes core surgical residencies to train clinical patient-care oriented surgeons and advanced residencies in such areas as thoracic surgery, cardiovascular surgery, pediatric surgery, and digestive organ surgery. Residents who want to become general thoracic surgeons must complete 350 cases of general surgery including at least 120 cases for an operator in the first 4 or more years in core programs. After passing the board for general surgery, they can advance to a subspecialty program for thoracic surgery in the final 3 or more years to pass the board for general thoracic surgery. It is essential to train the general thoracic surgeons who empathize with patients with thoracic diseases and can contribute to overcoming thoracic diseases by practicing evidence- or science-based medicine. PMID- 12722672 TI - [Introduction of new payment system (DRG/PPS) to university hospitals]. PMID- 12722673 TI - [Translational research for cancer therapeutics]. AB - The importance and necessity of promoting translational research in Japan are well recognized. Successful development of molecular cancer therapeutics is strongly dependent on the promotion of such translational research. Obviously, the success of translational research in the field of molecular cancer therapeutics is essential for the development of new drugs for cancer treatment in the 21st century. For this purpose, the formation of research teams composed of basic molecular biologists, basic pharmacological scientists and clinical physicians is important. At the same time, the research must be conducted on a scientific basis with the reliable and speedy development of supporting systems in Japan. PMID- 12722674 TI - [Brain tumors]. AB - Brain tumors generally arise as the culmination of a multistep process that involves a variety of genetic abnormalities. Theoretically, replacement of abnormal genes with normal genes is essential to brain tumor treatment. However, it is very difficult to replace all damaged genes. Currently, most clinical protocols for gene therapy in brain tumors include transfer of a gene which can induce tumor cells to die or which can enhance the environment to generate a systemic immune response against the tumor. The former strategy includes suicide gene therapies, tumor suppressor gene therapy and oncolytic virus therapy. The latter adopts immunogene therapy. In this report, we also focus on other gene therapies, such as therapies to control the cell cycle or apoptosis, and promote antiangiogenesis. Gene therapy is generally accepted to be rather safe in recent years. In fact, the current single-gene therapies for brain tumor are limited and probably restricted to a few tumors. Several agents with different mechanisms of action would be necessary to kill heterogenously mixed tumor cells. Further molecular techniques and basic studies may overcome the malignancy of cancers. PMID- 12722675 TI - [Adenoviral p53 gene therapy for human lung cancer]. AB - Recent advances in molecular biology have fostered remarkable insights into the molecular basis of neoplasms. This new understanding of cancer pathogenesis suggests that restoration of the function of critical gene products could halt or reverse these mechanisms, thus having a therapeutic effect in cancer. The tumor suppressor p53 gene has been implicated in many inherited and sporadic forms of malignancy in humans. A number of preclinical experiments have demonstrated that restoration of the wild-type p53 function in the cancer cell by gene transfer is sufficient to cause antitumor effects such as cell-cycle arrest and induction of apoptosis. This approach has entered initial clinical testing and provided intriguing information about the intratumoral administration of an adenovirus vector expressing the wild-type p53 gene in non-small cell lung cancer patients. PMID- 12722676 TI - [Gene therapy for breast cancer]. AB - Breast cancer is sensitive to chemotherapy and endocrine therapy, but the prognosis of advanced or relapsed breast cancer is unsatisfactory. Gene therapy is promising as another useful therapeutic approach for advanced breast cancer. Strategies of gene therapy for breast cancer in ongoing clinical protocols can be divided into four: (1) suppression of oncogenes or transduction of tumor suppressors; (2) enhancement of immunological response to cancer cells; (3) transduction of suicide genes; and (4) protection of bone marrow using drug resistance genes. We have started a clinical study of gene therapy for breast cancer using multidrug resistance gene (MDR1), in which advanced or relapsed breast cancer patients received high dose chemotherapy and autologous peripheral blood stem cell transplantation (PBSCT) with MDR1-transduced hemopoietic cells, and then were treated with docetaxel. Two patients have been treated so far, and in vivo enrichment of MDR1-transduced cells with docetaxel treatment after PBSCT was seen in both cases. Both patients are in complete remission and have no apparent adverse effect from MDR1 gene transduction. PMID- 12722677 TI - [Hematopoietic malignancies and gene therapy]. AB - In the case of hematopoietic malignancies, direct approach of gene therapy [gene transfer to cancer cells in order to obtain direct therapeutic effects (cell damage)] is difficult, because malignant cells are distributed in the whole body. As for indirect approaches, immuno-gene-therapy is investigated: As a unique approach, suicide gene therapy is applied to donor lymphocyte infusion for relapsed leukemia after bone marrow transplantation. The purpose of gene therapy is to eliminate donor lymphocytes quickly when severe side effects (GVHD) appeared. HSV-TK gene is generally utilized as a suicide gene. Basic studies are conducted to determine whether anti-tumor-angiogenesis therapy is also effective for hematological malignancies. In addition, leukemia development in 2 patients with X-linked severe combined immunodeficiency who underwent hematopoietic stem cell gene therapy is currently a serious problem in the field of gene therapy. In both cases, LMO2 gene was activated through insertional mutagenesis which was caused by retroviral vectormediated gene transfer. This genetic event is considered to be a trigger of T-lymphocytic leukemia development. Further basic studies are needed in terms of safety for stem cell gene therapy. PMID- 12722678 TI - [Usefulness of gemcitabine in the treatment of advanced pancreatic cancer in terms of its clinical benefits]. AB - Since pancreatic cancers are frequently discovered in an advanced stage, the expectations for chemotherapy are high. However, the response rate to chemotherapy of advanced cancer remains low. In Western countries, gemcitabine (GEM), a newly approved drug in Japan, was reported to be useful in improving symptoms and prolonging survival. In the present study, we compared 13 patients with unresectable advanced pancreatic cancer who were treated with GEM alone and 13 patients with pancreatic cancer treated with 5-fluorouracil/cisplatin (FP) as historical controls, with reference to therapeutic results, adverse reactions, and clinical benefit response. Although the response rate to GEM was low, at 7.7% (1/13), prolonged NC was seen in 76.9% (10/13) of cases, suggesting a contribution to prolonged survival (p = 0.0025). A clinical benefit response was seen in 38.5% (5/13) of the GEM group, as compared with the lower rate of 7.7% (1/13) in the FP group (p = 0.063). Since GEM is convenient to-administer, and does not diminish a patient's QOL, it is considered to be useful as a drug of first choice in the treatment of advanced pancreatic cancer in terms of clinical benefits. PMID- 12722679 TI - [Clinical study of TS-1 therapy for inoperable and recurrent gastric cancer]. AB - The clinical efficacy and safety of TS-1 therapy were studied retrospectively in patients with inoperable and recurrent gastric cancer. The subjects were 45 patients who were treated with TS-1 for more than 4 weeks at our center between May 1999 and July 2002. The objective overall response rate was 32% (14/44; 95% confidence interval, CI, 19-48). The response rate in the chemo-naive patients was 44% (11/25; 95% CI, 24-65), and that in the patients with previous chemotherapy was 16% (3/19; 95% CI, 3.4-40). Although doses or durations of TS-1 administration were reduced in 22 patients (reduction group) due to adverse effects or poor performance status, they achieved a fairly high response rate of 38% (8/21). For primary lesions, the response rate was 30% (8/27). The prevalence of adverse reactions with a grade of 3 or 4 was 36%. However, the prevalence of each grade 3 or 4 adverse effect was relatively low, at 13% for neutropenia, and around 5% for anorexia, nausea, vomiting, and diarrhea. The median administration period was 10 weeks (4-47 weeks) in all patients and 11 weeks (6-47 weeks) in the reduction group. The relative dose intensity was 0.89 in all patients and 0.81 in the reduction group. In patients who were treated until August 2001, the median survival time (MST) was 13 months with 1-year and 2-year survival rates of 53% and 14%, respectively. These results were similar to those reported in the phase II study for the new drug approval. This study demonstrated the reproducible activity and safety of TS-1 in practice. PMID- 12722680 TI - [Thymidine phosphorylase is correlated with DPD in colon cancer]. AB - Analysis of thymidine phosphorylase (TP), thymidylate synthase (TS) and dihydropyrimidine dehydrogenase (DPD) or their mRNA are now being applied before the start of chemotherapy to predict the therapeutic efficacy. Although these key enzymes were reported to be basically independent, we found the differences in TS between cancer and adjacent mucosa was reversely correlated with the difference in DPD. We found a significant relationship between TP and DPD in 52 patients with colon cancer. TP and DPD were measured by EIA. Statistical analysis was made non-parametrically using Statview 5.0. TP was significantly higher in cancer (78 +/- 58 U/mg protein) than in the adjacent mucosa (43 +/- 24 U/mg protein). Conversely, DPD was significantly lower in cancer(43 +/- 32 U/mg protein) than in the adjacent mucosa (55 +/- 18 U/mg protein). The amount of TP and DPD in cancer were not correlated with the clinicopathological parameters. TP was significantly (r = 0.70) correlated with DPD in cancer but not in adjacent mucosa. The difference in TP between cancer and adjacent mucosa was significantly (r = 0.69) correlated with the difference in DPD as well. In the colon cancer with low TP, DPD in cancer is lower than in the adjacent mucosa, however, the more TP in cancer increased, the higher the DPD in cancer increased over that in the adjacent mucosa. Modulation of DPD as well as TP may be necessary when high levels of TP or DPD are measured in the cancer tissue. The understanding of the basic relationship among these key enzymes will improve the 5-FU-based chemotherapeutic prediction. PMID- 12722681 TI - [Clinical outcome of intra-hepatic arterial infusion therapy for multiple liver metastases from colorectal cancer]. AB - Weekly infusion of high-dose 5-FU through the hepatic artery after resection of colorectal cancer was compared with resection alone in patients with multiple liver metastases. Twenty-seven patients (Group I) underwent hepatic artery infusion chemotherapy and 74 patients (Group II) underwent resection alone. The average survival time was 17.9 months in Group I and 8.5 months in Group II. One CR and 7 PR were achieved with arterial infusion therapy and the response rate was 29.6%. One- to three-year survivals were better in Group I than Group II. PMID- 12722682 TI - [Concurrent weekly nedaplatin-based radiotherapy for high risk, recurrent and advanced cervical cancer]. AB - Advanced cervical cancer has been predominantly treated with a combination of external beam and brachytherapy in Japan. Recent studies suggest concurrent use of cisplatin and radiation treatment has superior disease control to radiation only treatment. We have conducted a phase I pilot study of concurrent use of weekly nedaplatin (30 mg/m2) and sequential external beam and brachytherapy in advanced stage or recurrent uterine cervical cancer patients (n = 6). All patients completed the treatment without serious complications. Five patients had complete responses and one a partial response. The average AUC of nedaplatin after one administration was 5.0 micrograms/ml.hr. The therapeutic index was 2. We concluded that concurrent use of weekly nedaplatin and radiation is well tolerated by Japanese women, and may well be an excellent therapeutic modality for selected cases of advanced or recurrent cervical cancer. PMID- 12722683 TI - [Combination therapy of fluoropyrimidine (TS-1) administration and selective intra-arterial cisplatin infusion for tongue carcinoma--a case report]. AB - A 64-year-old male with primary squamous cell carcinoma of the tongue (T3N0M0) was treated with the novel fluoropyrimidine oral anticancer drug TS-1 and selective intra-arterial infusion of CDDP. His past history revealed pulmonary emphysema as a complication. Since his pulmonary function was reduced and it would have been difficult to perform surgery under general anesthesia, we started administration of TS-1 (100 mg/day) first and added selective arterial infusion of CDDP (5 mg/day) after one week of TS-1. On day 8 of TS-1 administration (day 2 of arterial infusion), the tumor had shrunk considerably, and on day 15 of TS-1 administration (day 9 of arterial infusion) the tumor had almost completely disappeared. Nausea and vomiting developed as adverse effects on day 20 of TS-1 administration (day 14 of arterial infusion), and administration of the anticancer drugs was stopped. Nutrition management and an antiemetic agent were started by intravenous drip infusion, and the adverse effects improved one week after administration was stopped. By the 5th week after the start of treatment, the tumor had disappeared macroscopically, and a CR had been achieved. Interstitial radiotherapy was performed as supplemental therapy, and as of this writing, September 3, 2002, 16 months after the start of treatment, the patient is being followed on an outpatient basis. His course has been favorable and recurrence-free. Although there were some slight adverse reactions, they were relatively mild, and since sufficient efficacy was observed, oral TS-1 plus selective intra-arterial CDDP therapy was concluded to be an effective method of treatment for patients with a past medical history making their general condition unfavorable. PMID- 12722684 TI - [Successful treatment with chemotherapy of vinorelbine and gemcitabine for an elderly patient with advanced non-small cell lung cancer]. AB - A 74-year-old man with non-small cell lung cancer was referred to our hospital for chemotherapy. On admission, he suffered from high fever and left upper back pain. Laboratory data showed marked leukocytosis and increased CRP. He was treated with chemotherapy of weekly vinorelbine and gemcitabine. After the second cycle, the levels of leukocytes and CRP were remarkably decreased, and the severe back pain was also alleviated. He has been given an additional 4 cycles as an outpatient. He is now doing well. This regimen seems to be effective in alleviating symptoms and improving QOL. PMID- 12722685 TI - [Delivery of twins after undergoing adjuvant LHRH analogue and CEF therapy]. AB - Chemotherapy is an important part of adjuvant therapy for younger patients with breast cancer, but prevention of ovarian dysfunction due to chemotherapy is important. There are some reports that pretreatment with LHRH analogue can preserve ovarian function. The present patient, who was treated in this way, delivered twins 6 years after surgery. A 33-year-old woman underwent breast conserving surgery for 1 cm carcinoma in the left A region, and pathologic findings were papillotubular carcinoma. t2n1 alpha (2/18) ER (+) PgR (+). LHRH analogue was administrated together with radiation therapy for the preserved breast. Six subsequent courses of CEF therapy were given, and then LHRH analogue was administered for 2 years and tamoxifen for 2 years and six months. Menstruation was recovered 6 months later and twins were delivered 3 years after the last administration of hormone therapy. This case should be referred in the treatment of young patients. PMID- 12722686 TI - [A case of gastric cancer with liver metastasis in which obstruction of the bile duct and choledocholithiasis was caused by intra-hepatic arterial infusion chemotherapy]. AB - A 64-year-old man underwent gastrectomy and partial liver resection for gastric cancer and liver metastasis, and was administered intra-arterial infusion chemotherapy for metastases of the remnant liver. This treatment was very effective against the liver metastases, but 13 months after the operation obstructive jaundice occurred. An examination revealed obstruction of the bile duct and choledocholithiasis. The choledocholithiasis was treated using a percutaneous transhepatic cholangio-scope, and choledocho-duodenostomy was performed for the obstruction of the bile duct. Findings from the operation suggested that the obstruction was caused by the intra-arterial infusion chemotherapy. At present, 2 years after the first operation, the patient is alive without the regrowth of the liver metastasis. PMID- 12722687 TI - [A case of advanced gastric cancer with peritoneal metastases responding to bi weekly paclitaxel therapy]. AB - A 68-year-old man underwent bypass operation for gastric cancer, because the tumor was judged to be unresectable due to peritoneal dissemination. After the operation, the patient was treated with daily oral administration of 100 mg TS-1 for two weeks followed by one week rest as one cycle. However, symptoms such as anemia, ascites and edema became worse and the TS-1 resulted in progressive disease. Bi-weekly paclitaxel therapy (80 mg/m2/2 weeks) was then chosen as second line chemotherapy. Anemia and edema were reduced and computed tomography showed shrinkage in the size of lymph nodes and disappearance of ascites. Only grade 1 alopecia was observed as an adverse event during the therapy. Bi-weekly paclitaxel therapy could be safe and useful as the second line therapy. PMID- 12722688 TI - [A case of advanced gastric cancer resected after successful treatment with the novel oral anticancer drug TS-1]. AB - We report a case of advanced gastric cancer resected after successful treatment with the novel oral anticancer drug TS-1. The patient was a 52-year-old male. Gastrointestinal fiberscopy showed advanced gastric cancer. Examinations by computed tomography revealed gastric cancer invasion of the pancreas and swollen para-aortic lymph nodes. This patient was treated by preoperative chemotherapy with oral administration of TS-1 (120 mg per day). After 3 courses of treatment of TS-1, the primary lesion and swollen lymph nodes were remarkably reduced. This chemotherapy enabled total gastrectomy in curative resection. The pathological effectiveness of chemotherapy was Grade 1b in the primary lesion and Grade 2 or 3 in the lymph nodes. The patient sustained few side effects. This preoperative chemotherapy regimen seems to be an effective and promising therapy for patients with advanced gastric cancer. PMID- 12722689 TI - [A case of irinotecan-resistant colon cancer responding to chronotherapy with oxaliplatin, 5-FU, l-LV]. AB - We report a case of irinotecan-resistant colon cancer responding to chronotherapy with oxaliplatin (L-OHP), 5-FU, l-LV (l-Leucovorin). A 72-year-old man was examined at a certain hospital because of constipation and appetite loss. Chest computed tomography (CT) revealed lung metastases, and abdominal CT revealed liver metastases. He was then referred to our hospital. A colonoscopy revealed type 2 tumor in the colon, at the hepatic flexure. We diagnosed adenocarcinoma of the colon with metastases to the liver and lung. Resection of the primary lesion was performed, and chemotherapy consisting of systemic administration of CPT-11, 5-FU and l-LV was performed. After 2 courses of combined treatment with CPT-11/5 FU/l-LV, CT revealed considerable reduction of the metastatic tumors. However, after 3 courses of combined treatment, progressive disease was observed and new brain and bone metastases were detected. We imported and used a non approved/pending drug, oxaliplatin from the Remedy and Health Corporation, with informed consent from the patient and his family and our clinical ethics committee. Chronotherapeutic schedules have been performed, from which the safety and activity of oxaliplatin against advanced colorectal cancer was reported. Our patient received a 5-day course of chronomodulated 5-FU and l-LV (750 and 300 mg/body/day, respectively; peak delivery rate at AM 4:00 hours) with L-OHP on the first day of each course (100 mg/body, as a 6-hour infusion). Each course was again repeated every 21 days. A partial response was observed in the liver and lung metastases. These results indicate that chronomodulated 5-FU and LV with L OHP therapy could be an effective regimen for cases of irinotecan-resistant colon cancer. PMID- 12722690 TI - [A case of para-aortic lymph node recurrence of gallbladder cancer completely responding to single drug (UFT) chemotherapy]. AB - A 67-year-old man with gallbladder cancer was treated by cholecystectomy and extrahepatic bile duct resection with regional lymph node dissection. At 10 months after surgery, CT demonstrated para-aortic lymph node recurrence. Single drug chemotherapy of UFT at 400 mg was started. After one month, the lymph node recurrence could not be detected by CT. UFT may be the primary candidate for chemotherapy for lymph node recurrence of gallbladder cancer. PMID- 12722691 TI - [Gemcitabine improved QOL and survival in a case of diffuse malignant pleural mesothelioma]. AB - Diffuse malignant pleural mesothelioma is a disease with poor prognosis, and no standard therapy for inoperable cases has been established. There are several reported trials using various anticancer drugs, but their sample sizes were small and none documented the drugs' effectiveness. Recently, some reports revealed that gemcitabine (GEM), which has been demonstrated to be effective for the treatment of non-small cell lung carcinoma, was effective in cases of diffuse malignant pleural mesothelioma. We also treated a 76-year-old female patient with inoperable, biphasic malignant pleural mesothelioma in whom monotherapy with GEM 1,000 mg/m2 was continued leading to improved QOL and survival for 2 years. Though diagnostic imaging indicated SD (stable disease), a high efficacy was observed in terms of reduced subjective symptoms such as a feeling of difficulty in breathing and pain. It is suggested that GEM is a potentially effective drug for the improvement of QOL in inoperable cases of diffuse malignant pleural mesothelioma. PMID- 12722692 TI - [Globalization of clinical trials]. AB - Based on reviews of the Japanese clinical trial situation in lung cancer, gastric cancer, prostate cancer and breast cancer, it was clear that much progress has been made in short time. There are considerable differences between Japan and the West and also differences between clinical areas in Japan. For regulatory purposes bridging studies have become increasingly important. Use of identical protocols are required for effective bridging. Participations in global phase III trials is the best way of achieving registration in Japan. For successful global trials in Japan it is important to include Japanese investigators in the preparation of the protocol and to recognise the challenges facing such a project. Clinical practice in diagnosis and treatment have many differences, thus it is recommended to have clear and detailed information in the protocol. Hard end points like survival are important since they are not biased by cultural differences. There are clear difficulties with HE or QOL outcomes. The emergence of focus on evidence based medicine is also happening in Japan and will help to harmonize documentation across the world. For large adjuvant or prevention cancer global trials are essential. To facilitate global studies further development of infrastructure is necessary in Japan. Use of electronic data capture web based communication etc. will help overcome communication difficulties. Other improvements that will make Japanese participation in global trials easier and better include establishment of clinical trial centre at each hospital, introduction of trial coordinators or study nurses and an improved collaboration with company staff. A critical issue that also need addressing is agreement of centre target recruitment. We need to introduce a new flexible system in Japan if participation in global trial is to be optimised. If we can address these issues Japanese investigators and collaborative groups should be able to initiate and lead global trials in the future. PMID- 12722693 TI - Sudden death due to atrial fibrillation in hypertrophic cardiomyopathy: a predictable event in a young patient. AB - This case refers to a 39-year-old woman with hypertrophic cardiomyopathy (HCM) and family history of sudden death (SD). In 1985, high rate atrial stimulation induced VF. In 1996 an ICD was implanted and she remained without arrhythmic events until November 2000 when the device reported one episode of atrial fibrillation degenerating into VF and terminated by the ICD. The VF induction mechanism recorded by the ICD was similar to that observed in 1985. The high incidence of atrial tachyarrhythmias in HCM renders cases like this at higher risk of SD. The predictive role of incremental atrial stimulation merits highlighting in future studies. PMID- 12722694 TI - Median nerve somatosensory evoked potentials and upper limb motor function in hemiparesis. AB - The purpose of this study is to describe the relationship between median nerve somatosensory evoked potential (SEP) parameters and clinical measures of motor impairment and physical disability in the affected upper limb of patients with hemiparesis. SEP assessments were carried out in 28 long-term survivors of stroke. Stroke survivors with preserved SEPs over the affected hemisphere exhibited significantly lower motor impairment and physical disability than those with absent waveforms. Among those with preserved waveforms, SEP amplitude exhibited a weak correlation with degree of upper limb motor impairment. The correlation between SEP amplitude and physical disability was not statistically significant. Contrary to prior reports, median nerve SEP appears to have a weak correlational relationship with clinical measures of upper limb function in hemiparesis. PMID- 12722701 TI - How Buddhism influences pain control choices. PMID- 12722703 TI - [Exercise training in heart failure]. AB - BACKGROUND: In patients with chronic heart failure, exercise limitation cannot be exclusively attributed to cardiac dysfunction. During progression of the disease, peripheral factors have a growing impact on functional capacity. Chronic underperfusion of skeletal muscles, inactivity and a harmful effect of neuroendocrine stimulation lead to a myopathy syndrome characterized by atrophy, a shift in muscle fiber type and a loss of capillaries and mitochondria, which cause reduced muscle strength and aerobic capacity. Unlike left ventricular systolic function, which is a poor predictor of exercise tolerance in patients with chronic heart failure, parameters characterizing the myopathy syndrome correlate well with functional capacity. In the last 20 years, numerous studies have demonstrated that chronic heart failure patients in NYHA classes II and III benefit from physical training without deterioration of their cardiac function. EFFECTS OF EXERCISE TRAINING: Physical activity primarily improves muscle function and increases maximal as well as submaximal exercise capacity. Advantageous effects have also been observed with regard to neuroendocrine stimulation. The effect on central hemodynamics is much smaller and indirectly mediated by a trend towards decreased peripheral resistance. Left ventricular diameters, ejection fraction and filling pressures do not show significant changes. The gain in exercise capacity is usually accompanied by an improvement in symptom status and quality of life. Recent studies indicate that prognosis may also be improved by exercise training. CONVERSION INTO CLINICAL PRACTICE: Although continuous exercise training (mainly by use of cycle ergometry) has been investigated most thoroughly, there is an increasing number of studies reporting beneficial effects of interval and resistance training. Before starting a standardized training program, functional limitation and training intensity should be determined by an appropriate stress test. During the first training sessions, patients should always be supervised by a physician. In order to increase efficiency and feasibility of the exercise therapy, home-based training should be engaged as soon as a stable condition during repeated training sessions has been proven. PMID- 12722706 TI - [Expert consultant emphasizes more competition in health care]. PMID- 12722709 TI - Nurses and needlesticks, then and now. PMID- 12722707 TI - [Politics and medicine in the 21st century--misunderstandings, insolvency or foolishness?]. PMID- 12722710 TI - [Cardio-MR and cardio-CT: quo vadis?]. PMID- 12722713 TI - Starting peripheral I.V.s: tips for planning ahead. PMID- 12722714 TI - AIDS/HIV roundup. PMID- 12722715 TI - 50th anniversary of the first ascent of Everest. PMID- 12722718 TI - [Gradual progress towards optimal treatment of diabetes]. PMID- 12722721 TI - Managing a patient with a venous ulcer. PMID- 12722720 TI - Closing in on clues to encephalitis. PMID- 12722722 TI - How do I manage a spinal epidural hematoma? PMID- 12722723 TI - Protecting your patient during a seizure. PMID- 12722724 TI - Myths and facts...about ephedrine use. PMID- 12722725 TI - Detecting neuromuscular problems with electromyography. PMID- 12722726 TI - You routinely administer combination drugs... PMID- 12722728 TI - [On the 100th birthday of Ernst Jeckelin]. PMID- 12722729 TI - Healing and feeling: the clinical ontology of emotion. AB - In the clinical setting, not enough attention is paid to the role that emotion plays. It is at worst ignored or avoided, isolating those who are suffering, at best treated as something to help another to endure. This is the result, in part, of an impoverished idea that views emotion as mere feelings. However, emotions are not just feelings, they are cognitive. If we look beneath the surface, emotions can provide information about values and beliefs, some of which may be false or unreasonable. When they are motivating decisions that seriously affect the lives of others, there is a moral obligation to ferret them out and correct or deal with them. Attention needs to be paid to the emotions of healthcare providers as well as to the emotions of patients and their loved ones, and more done to train healthcare providers to deal effectively and productively with emotion--their own and others'. A son tearfully requests everything be done for his dying father. He fears limiting treatment means his father will be ignored. A physician orders another course of chemotherapy because she cannot bear for this patient to die. The patient, fearing he will painfully suffocate to death, agrees. Paying attention to emotions and bringing to light the beliefs and values behind them can only enhance the care and consideration of all involved. PMID- 12722731 TI - Editorial: simulation--saviour or Satan? PMID- 12722730 TI - The conjoined twins and the limits of rationality in applied ethics. AB - In this article I consider the case of the surgical separation of conjoined twins resulting in the immediate and predictable death of the weaker one. The case was submitted to English law by the hospital, and the operation permitted against the parents' wishes. I consider the relationship between the legal decision and the moral reasons adduced in its support, reasons gaining their force against the framework of much mainstream normative ethical theory. I argue that in a few morally dilemmatic situations, such a legalistic-theoretical approach cannot plausibly accommodate certain irreducible and ineliminable features of the ethical experience of any concrete individual implicated in the situation, and that this failure partly undermines its self-appointed role of guiding such an individual's conduct. For example, the problem as experienced by the judge and by the parents might not be the same problem at all, and some of their respective reasons may be mutually unintelligible or impotent. I certainly do not argue for a rejection of law or of moral theory; I merely challenge their implicit claim to comprehensiveness and their fixation with an idealised and putatively universal rationality modelled on converging scientific enquiry. Finally, I claim that at least in the twins' case there may be insufficient normative robustness to the conclusions reached, or indeed reachable, by the court in a situation where intuitions and moral reasons pull in fundamentally incommensurable directions; as such, there may be room for an acknowledgement of the spiritual, through a humble abstention from making a decision--which is not to be confused with deciding to do nothing. PMID- 12722732 TI - [Antiarrhythmic drugs in atrial fibrillation]. PMID- 12722733 TI - Navigating the paraneoplastic neurological syndromes. PMID- 12722734 TI - The problematic role of 'irreversibility' in the definition of death. AB - Most definitions of death--whether cardiopulmonary, whole brain and brain stem, or just upper brain--include an irreversibility condition. Cessation of function is not enough to declare death. Irreversibility should be limited to an organism's ability to 'restart' itself after vital organs have ceased to function. However, this would mean that every hour people who cannot be revived without the intervention of medical personnel and their technology are coming back from the dead. However, the alternative of irreversibility being dependent upon technology will lead to even more counterintuitive results such as: some people are dead at a particular time and place, but others in more technologically advanced eras and locations are alive despite their being in identical physical states; in the future, millions of cryogeneically frozen human beings could spend centuries in a non-dead state because of the future technological breakthroughs; or large numbers of 'frozen' people are dead for aeons but coroners are not able to declare them so because they are unaware of what biological conditions science will never be able to reverse. So death should be defined only in non-relational biological terms, with a self-starting condition similar to that once advocated by Lawrence Becker. PMID- 12722735 TI - 5-HT(1)-like receptor agonists and the pathophysiology of migraine. AB - Recently, AG-25086 and GR-43175 have been reported to be highly effective in aborting acute migraine attacks. These compounds seem selectively to stimulate a sub-population of 5-HT(1)-like receptors to inhibit noradrenaline release from certain sympathetic neurovascular terminals, to contract dog saphenous vein and dog, monkey and human basilar arteries, and to decrease the arteriovenous anastomotic component of carotid blood flow in the cat. GR-43175 neither has any antinociceptive effect nor crosses the blood-brain barrier. Pramod Saxena and Michel Ferrari review the clinical effectiveness of these 5-HT(1)-like receptor agonists and their selective pharmacology, both of which strongly suggest that excessive dilatation in the extracerebral cranial (scalp and/or dural) vasculature is an integral part of the pathophysiology of migraine. PMID- 12722736 TI - Information giving in clinical trials: the views of medical researchers. AB - It is both an ethical and legal requirement that patients who participate in clinical trials must generally give their consent. As part of this process, patients must be provided with adequate information to enable them to decide whether or not to take part. In the UK, the pharmaceutical companies that sponsor such research, as well as Local Research Ethics Committees, specify in detail the information that must be given to trial participants. The researchers who conduct clinical trials inevitably form views on the amount of information they are required to provide, and about patients' comprehension of that information. The literature in this area suggests that some medical researchers may be unhappy with the amount of information that they must give patient participants. There have been, however, few systematic attempts to determine their views. This paper reports a study that explored researchers' views as to (i) the amount of information provided to trial participants, and (ii) participants' understanding of that information. Researchers generally felt that they were required to give trial participants an appropriate amount of information, and that most patients had at least a reasonable understanding of key aspects of the clinical trials' process. However, there were differing views as to the level of information that they felt patients themselves wanted. The researchers did not generally feel that the patients' inability to comprehend information rendered the process of obtaining 'informed consent' a waste of time. However, some did believe that they were required to burden patients with excessive information. PMID- 12722738 TI - Proceedings of the 6th Biennial Motor Control and Human Skill Research Workshop. Fremantle, Western Australia, December 2001. PMID- 12722737 TI - New horizons in biotechnology. Proceedings of an international conference. April 18-21, 2001. Trivandrum, India. PMID- 12722739 TI - 125I-BMIPP and 18F-FDG uptake in a transgenic mouse model of stunned myocardium. AB - Reported metabolic patterns in myocardial stunning are not uniform. We investigated relative myocardial perfusion, glucose and fatty acid uptake using a technetium-99 hexakis-2-methoxyisobutyl-isonitrile (MIBI), fluorine-18 2-fluoro-2 deoxyglucose (FDG) and iodine-125 15-(p-iodo-phenyl)-3(R,S)-methylpentadecanoic acid (BMIPP) mixture, in a recently developed transgenic (TR) mouse model which mimics stunned myocardium. Twenty-seven mice - 14 TR and 13 age-matched wild type controls (C) - were divided into four groups: TR-fed, TR-fasted, C-fed and C fasted. Animals were sacrificed 2 h after injection, tissue samples counted and percent-injected dose/gram tissue (% id/g) calculated for each radioisotope. Tissues were also Folch extracted and 125I incorporation into the various lipid pools (TG, triglycerides; DG, diglycerides; FFA, free fatty acids; PL, phospholipids) was determined by thin-layer chromatography (TLC). The pooled data for each of the four groups (TR-fed vs C-fed and TR-fed vs C-fasted) showed no differences in myocardial blood flow (% MIBI id/g), glucose uptake (% FDG id/g) or fatty acid uptake (% BMIPP id/g). Only minor differences were observed in the incorporation of 125I-BMIPP into the myocardial TG, DG, FFA and PL lipid pools. However, significantly decreased myocardial FDG uptake was observed in a subset of fasted mice - four out of ten TR-fasted mice (3.4% vs 20.5% id/g) and three out of nine C-fasted mice (5.5% vs 30.6% id/g). The transgenic mouse model of stunned myocardium shows normal myocardial perfusion and overall intact myocardial glucose and myocardial fatty acid uptake as determined with clinically applicable radiolabelled analogues. These data are in line with the hypothesis that the contractile inefficiency in stunned myocardium is not linked to metabolic alterations but is associated with an insufficient chemical to mechanical energy coupling. PMID- 12722741 TI - 111In-oxine and 99mTc-HMPAO labelling of antigen-loaded dendritic cells: in vivo imaging and influence on motility and actin content. AB - In cancer vaccination trials, antigen-loaded dendritic cells (DCs) are usually injected intradermally and are expected to rapidly move to a regional lymph node where antigen presentation should occur. In this study we investigated the influence of indium-111 oxine (111In) and technetium-99m hexamethylpropylene amine oxime (99mTc-HMPAO) labelling on the motility and actin content of antigen loaded DCs in parallel with in vivo migration in humans. Human autologous monocyte-derived DCs loaded with a tumour antigen were labelled with 111In (0.11, 0.37 or 0.74 MBq/10(7) DCs) or 99mTc-HMPAO (18.5 or 185 MBq/10(7) DCs). 111In labelling was much more stable than 99mTc-HMPAO labelling. Quantitative videomicroscopy showed that the mean distance of displacement of DCs increased in accordance with the 111In activity used for labelling. Monomeric (G) and filamentous (F) actin content of DCs evaluated by quantitative immunofluorescence demonstrated that the ratio of filamentous to globular actin content in labelled DCs increased significantly in accordance with the activity used for labelling with both tracers. Twelve patients enrolled in a phase I/II vaccination trial received injections of 10(7) antigen-loaded DCs labelled with either 0.74 MBq of 111In (group A, n=6/12) or 18.5 MBq of 99mTc-HMPAO (group B, n=6/12) in the proximal part of the legs, one intradermally on one side, one subcutaneously on the opposite side. In three of the six patients of each group, antigen-loaded DCs were incubated with monophosphoryl lipid A (MPL) just before the labelling, in order to initiate the maturation process (subgroup MPL+). Only one MPL+ patient of group A exhibited faint focal uptake in the inguinal region on the late images. Group B presented a more complex pattern of radioactivity distribution (early bladder activity without brain uptake) indicating that 99mTc-HMPAO is not a suitable radiopharmaceutical for labelling of loaded DCs. The activity cleared from DCs as a labelled molecule different from the lipophilic 99mTc-HMPAO. Only one of the six patients had nodular inguinal uptake on the intradermally injected side (DCs not incubated with MPL). In conclusion, the present study did not demonstrate migration of loaded labelled DCs from intradermal or subcutaneous sites of injection to regional lymph nodes. This provides an indication that a large proportion of antigen-loaded DCs, as used in current human trials for cancer therapy, may not reach regional lymph nodes. PMID- 12722742 TI - MRI monitoring of tumor response following angiogenesis inhibition in an experimental human breast cancer model. AB - The aim of this study was to evaluate the potential of dynamic magnetic resonance imaging (MRI) enhanced by macromolecular contrast agents to monitor noninvasively the therapeutic effect of an anti-angiogenesis VEGF receptor kinase inhibitor in an experimental cancer model. MDA-MB-435, a poorly differentiated human breast cancer cell line, was implanted into the mammary fat pad in 20 female homozygous athymic rats. Animals were assigned randomly to a control (n=10) or drug treatment group (n=10). Baseline dynamic MRI was performed on sequential days using albumin-(GdDTPA)30 (6.0 nm diameter) and ultrasmall superparamagnetic iron oxide (USPIO) particles (approximately 30 nm diameter). Subjects were treated either with PTK787/ZK 222584, a VEGF receptor tyrosine kinase inhibitor, or saline given orally twice daily for 1 week followed by repeat MRI examinations serially using each contrast agent. Employing a unidirectional kinetic model comprising the plasma and interstitial water compartments, tumor microvessel characteristics including fractional plasma volume and transendothelial permeability (K(PS)) were estimated for each contrast medium. Tumor growth and the microvascular density, a histologic surrogate of angiogenesis, were also measured. Control tumors significantly increased (P<0.05) in size and in microvascular permeability (K(PS)) based on MRI assays using both macromolecular contrast media. In contrast, tumor growth was significantly reduced (P<0.05) in rats treated with PTK787/ZK 222584 and K(PS) values declined slightly. Estimated values for the fractional plasma volume did not differ significantly between treatment groups or contrast agents. Microvascular density counts correlated fairly with the tumor growth rate (r=0.64) and were statistically significant higher (P<0.05) in the control than in the drug-treated group. MRI measurements of tumor microvascular response, particularly transendothelial permeability (K(PS)), using either of two macromolecular contrast media, were able to detect effects of treatment with a VEGF receptor tyrosine kinase inhibitor on tumor vascular permeability. In a clinical setting such quantitative MRI measurements could be used to monitor tumor anti-angiogenesis therapy. PMID- 12722745 TI - Abstracts from the 8th European ISSX Meeting. April 27-May 1, 2003. Dijon, France. PMID- 12722743 TI - [Re the review article by V. Votava, L. Svobodova, J. Homolka: Interstitial Pulmonary Processes]. PMID- 12722746 TI - Abstracts of the 25th Annual Meeting of the Japan Neuroscience Society. Tokyo, Japan, July 7-9, 2002. PMID- 12722747 TI - Medicaid matters: children's health and Medicaid eligibility expansions. AB - In the late 1980s, a series of federal laws were enacted which expanded Medicaid eligibility to more of the nation's children. States had a great amount of discretion in how fast and how far these expansions were implemented. As a result, there was great variation among the states in defining who was eligible for the program. This variation provides a rare opportunity to disentangle the effect of Medicaid from a child's socioeconomic status. Using data from the National Health Interview Survey, we address whether the Medicaid expansions improved the health and functional status of children. Econometric models were developed using fixed-effects regressions, and were estimated separately for white, black, and Hispanic children. White children experienced statistically significant reductions in acute health conditions and functional limitations. Black and Hispanic children showed some evidence of improved health conditions and functional status, but this evidence is inconclusive in the study sample. This may be due to differences in their access to appropriate health services or to the smaller sample size of minorities in each geographic area. The findings are also relevant to the implementation of the Children' Health Insurance Program (CHIP), the latest federal effort to expand access to health care to poor and near poor children. In many states, CHIP is being implemented in whole or in part through further Medicaid expansions. PMID- 12722748 TI - Abstracts of the 58th Annual Meeting of the Society of Biological Psychiatry. May 15-17, 2003, San Francisco, California, USA. PMID- 12722750 TI - A new era in methods of embryo manipulation? PMID- 12722751 TI - Abstracts of the North American Society of Pacing and Electrophysiology 24th annual meeting. May 14-17, 2003. Washington, DC, USA. PMID- 12722752 TI - The value of reducing risk of death: a policy perspective. PMID- 12722753 TI - History of heart failure. PMID- 12722754 TI - Abstracts of the 13th Keio University International Symposium for Life Science and Medicine: Research and Development of Artificial Oxygen Carrier--It's Frontline. February 28-March 1, 2003. Tokyo, Japan. PMID- 12722755 TI - [Abstracts of the Congress of the 50th Anniversary of the Gastroenterology Society of Peru. 30 October-1 November 2002]. PMID- 12722757 TI - [Comments on the German brief version of the Wender Utah Rating Scale]. PMID- 12722756 TI - [Neuropsychiatry and parapsychology of autoscopic phenomena]. PMID- 12722758 TI - [Ernst Grunthal (1894-1972). Remarks on the cover illustration]. PMID- 12722759 TI - [Psychopharmacological regulations. Results and commentary to the report on drug regulations 2002]. PMID- 12722760 TI - Innovations and intellectual property: the case of genomic patenting. AB - In an effort to balance static and dynamic efficiency in the production and use of knowledge, societies institute intellectual property policies. In the United States, the patent system is a well-established mechanism to provide inventors with time-limited protection of new technologies in exchange for disclosure of information about their inventions. Emerging biotechnology, specifically the filing of patents on gene sequences, raises serious questions about whether the patent system is appropriately weighing societal costs and benefits in its grants of intellectual property protection. Gene sequences represent a hybrid case between discrete inventions and more general pieces of information that are useful for many, potentially very different, purposes. This information content in genes makes it possible for a patent on a gene to cover a wide range of possible technological applications and, as a result, be of unknown breadth when issued. This analysis explores the potential effect of these characteristics on future innovation in biotechnology. PMID- 12722761 TI - [Ethics in tension center of postmodern antagonisms]. PMID- 12722763 TI - Abstracts of the European Congress of Radiology. March 7-11, 2003. Vienna, Austria. PMID- 12722762 TI - Hospital selective contracting without consumer choice: what can we learn from Medi-Cal? AB - In the selective contracting era, consumer choice has generally been absent in most state Medicaid programs, including California's (called Medi-Cal). In a setting where beneficiary exit is not a threat, a large payer may have both the incentives and the ability to exercise undue market power, potentially exposing an already vulnerable population to further harm. The analyses presented here of Medi-Cal contracting data, however, do not yield compelling evidence in favor of the undue market power hypothesis. Instead, hospital competition appears to explain with greater consistency why certain hospitals choose to contract with Medi-Cal while others do not, the trends in inpatient prices paid by Medi-Cal over time, and the effect of price competition on service cutbacks, such as emergency room closures. PMID- 12722764 TI - [Postoperative infections in implanted port systems?]. PMID- 12722765 TI - [Pancreatic carcinoma--overcome technically but not oncologically]. PMID- 12722766 TI - Strategy for e-PTFE separation. PMID- 12722768 TI - Proceedings of the First International Union of Biochemistry and Molecular Biology/South African Society for Biochemistry and Molecular Biology (IUBMB/SASBMB) Special Meeting on Biochemistry and Molecular Basis of Disease. Cape Town, South Africa, November 19-23, 2001. PMID- 12722767 TI - Microalbuminuria in insulin-dependent diabetes mellitus always indicative of diabetic nephropathy? PMID- 12722769 TI - Glaucoma is present prior to its detection with standard automated perimetry: is it time to change our concepts? PMID- 12722770 TI - Preoperative biometry of cataractous eyes using partial coherence laser interferometry. PMID- 12722771 TI - Bibliography. Current world literature. Maternal-fetal medicine. PMID- 12722772 TI - Bibliography. Current world literature. Prenatal diagnosis. PMID- 12722773 TI - Current trends and controversies in ischemia-reperfusion research--meeting report of the Hatter Institute 3rd International Workshop on Cardioprotection. PMID- 12722774 TI - Interview with Louis W. Sullivan, M.D., on his early childhood years. Interview by Clarice G. Conley. PMID- 12722775 TI - Inappropriate attribution: the "Lazy Author Syndrome". PMID- 12722776 TI - Bibliography. Current world literature. Infectious diseases. PMID- 12722777 TI - Immediate repair compared with surveillance of small abdominal aortic aneurysms. PMID- 12722778 TI - [70th anniversary of the St. Petersburg I.I. Dzhanelidze Emergency Medical Care Institute]. PMID- 12722779 TI - Medical errors and their prevention. PMID- 12722780 TI - The care of children from ethnic minorities in our hospitals. PMID- 12722781 TI - Patients presenting with acute myocardial infarction to a district general hospital: baseline results and effect of audit. AB - The Cardiovascular Health Strategy recommended that patients presenting with acute myocardial infarction receive thrombolysis within ninety minutes of alerting medical or ambulance services. The aim of this prospective study was to describe the management of patients with acute myocardial infarction (AMI) presenting to a district general hospital in Donegal. All patients with a confirmed diagnosis of acute myocardial infarction, excluding those from the Donegal Area Rapid Treatment Study (DARTS) practices, admitted to Letterkenny General Hospital (LGH) from 31.08.99 to 31.08.01 were included in the study. 349 patients were included in the study; average age of 68 ranging from 30 to 96 years and 69% were male. Of the 349 patients, 101 (29%) were located more than 30 miles from LGH at the time of onset of symptoms. The median time taken from the onset of symptoms to calling for help was 119 minutes. The median time from hospital arrival to patients being admitted to CCU was 90 minutes. Thrombolytic therapy was administered in 31% of patients; for these patients the median call to needle time was 200 minutes. Call to needle times differed significantly between rural (median 227.5 minutes n = 64) and urban patients (median 175 minutes n = 37, p < 0.05, Mann-Whitney). Hospital delay times decreased throughout the study period (p > 0.05, Mann-Whitney). The study extends the findings from previous research by investigating the individual time delay components from onset of symptoms to treatment in AMI patients. Delay times exceed the recommended call to needle and door to needle times suggesting the need for interventions to reduce these times. PMID- 12722782 TI - An investigation of the psychosocial impact of a Compensation Tribunal on women with latrogenic hepatitis C infection. AB - The aim of this study was to investigate the psychosocial impact of a Compensation Tribunal in women with an iatrogenic hepatitis C virus (HCV) infection. Eighty-three women diagnosed with an iatrogenic HCV infection were recruited, 19 women were Pre-Tribunal and 64 women were post-Tribunal. Both standardised and disease specific psychological measures were used. A series of t tests revealed no differences in psychological well-being and adjustment to HCV infection in women pre and post Compensation Tribunal. Chi-square tests revealed no association between PCR status and 1) psychological well-being and 2) experience of anger/blame in women post-Compensation Tribunal. A further series of t-tests revealed that women with high levels of anger and blame post Compensation Tribunal perceived their future as more uncertain, experienced more pain, low self-esteem and psychological distress, viewed their ability to work as impeded and complained of increased stress preparing for their Compensation Tribunal. This study suggests that poor adjustment in women with an iatrogenic HCV infection post-Compensation Tribunal is not associated with attendance at a Compensation Tribunal nor PCR status but rather to experiences of anger and blame. PMID- 12722783 TI - A comparison of the effectiveness of pre-hospital bag-valve-mask ventilation performed by Irish emergency medical technicians and anaesthetists working in a tertiary referral teaching hospital. AB - The objective of this study was to assess the effectiveness of bag-valve-mask ventilation performed by emergency medical technicians with prehospital clinical responsibilities and to compare this with anaesthetists working clinically in tertiary referral teaching hospitals. Participants were asked to perform bag valve-mask ventilation for three minutes on a Resusi Anne manniquin using a facemask and a two litre self inflating bag. The tidal volumes generated were quantified using a Laerdal skillmeter computer as grades 0-5 corresponding to 0, 335, 434, 561, 673 and > 800 mls. The effectiveness of bag-valve-mask (i.e. the proportion of ventilation attempts which achieved a tidal volume of > 434 mls) was greater for emergency medical technicians [88.2 (17.1)%] than for anaesthetists [40.4 (36.5)%] (P<0.001). Six of the 27 anaesthetists, but none of the 29 emergency medical technicians, were unable to produce even one effective tidal volume i.e. > 434 mls. It is likely that emergency medical technicians are able to perform adequate bag-valve-mask ventilation. PMID- 12722784 TI - Efficacy of pulsed-dye laser for viral warts--an internal audit. AB - Pulsed-dye laser (PDL) surgery is a recognized treatment of viral warts. Initial studies in 1993 by Tan et al achieved a 72% cure rate for 39 patients after a mean of 1.68 treatments. Since then, conflicting response rates ranging from 47 95% have been reported. The records of 44 patients, who attended the Laser Unit at St. James's Hospital between January 1999 to June 2000, were reviewed. Follow up was conducted by telephone interview or questionnaire. Twenty-eight (64%) patients identified for the study responded. The average number of treatments was 4 (range 1-12) and mean treatment intervals were 4.9 weeks (3-10). Our results showed 64% complete clearance of all the areas treated, and 46% complete clearance of the 13 areas identified as recalcitrant. Rates for complete clearance varied with anatomic site: 73% for the face, 68% for the hands, and 40% for feet. Seventy-eight percent (7/9) of periungal warts completely cleared, and 70% (7/10) mosaic warts cleared. Twenty-five percent of the patients complained of severe pain during treatment. Post-operative complications were few, and rarely affected the patients' lifestyle. The recurrence of warts, in weeks to months following the last treatment, was reported by 36% of the patients. Seventy nine percent of patients were satisfied with the treatment they were given. PDL may be useful in treatment of refractory warts in selected patient populations. The study identified a need to address pain control in a significant number of patients. PMID- 12722785 TI - Carbon monoxide poisoning in the Republic of Ireland. AB - Carbon monoxide (CO) poisoning remains a common preventable cause of morbidity and mortality from poisoning worldwide. Common environmental sources include motor vehicle exhaust, faulty kerosene or gas heaters used in unventilated rooms, and fireplaces with blocked flues. This paper describes the epidemiology of CO poisoning in Ireland, using Hospital Inpatient Enquiry (HIPE) data, Mortality data and data from the National Poisons Information Centre (NPIC). CO poisoning is responsible for over 40 deaths per year in Ireland. Many occur at home as a result of house-fires. Incomplete combustion of domestic fuels is responsible for many cases of non-fatal unintentional CO poisoning. In comparison, most intentional poisonings are caused by motor vehicle exhaust, representing approximately 6.4% of successful suicides. The advent of catalytic converters in cars may lead to a decline in the effectiveness of this method of suicide. This study illustrates the hidden impact of CO poisoning. Most deaths occur at home and thus do not come to the attention of the health services. Those admitted to hospital represent the milder end of the spectrum and generally recover after a short stay. It is important that the public be aware of the risks of CO poisoning and that Public Health action be taken. PMID- 12722786 TI - Thirty year trends in a large Irish obstetric cohort. AB - To evaluate sequential trends in respect of parity, maternal age, gestation at delivery and birthweight in a large cohort of the national birth total during the period 1968 to 1998. Retrospective review of obstetric variables in six biennial cohorts at six yearly intervals, among mothers delivering at the National Maternity Hospital (NMH), Dublin. Where feasible comparisons have been made with contemporary national data. During the thirty year period, total NMH births varied between 11 and 14% of the Irish total. The proportions of primigravidae and grandmultiparae have increased by 15% and decreased by 21% respectively. There has been a shift in the predominant childbearing age group from 25-29 years to the 30-34 years. The proportion of teenage pregnancies has altered very little but births to women over 40 years has declined sharply from 6.0% to 3.0%. Births outside marriage have increased from 5% to 29%. Although, there has been a trend towards increasing average birthweight, no increase was observed in infants exceeding 4.5Kg. Total infants weighing under 1.5Kg have more than doubled. National birth statistics generally reflected NMH trends but with an apparent year time lag. PMID- 12722787 TI - The Football World Cup 2002--analysis of related attendances to an Irish Emergency Department. AB - Throughout the Football World Cup Finals of 2002, 47 patients attended the Emergency department with a variety of conditions related to the event. The majority were minor trauma victims such as assaults or falls and in over half the presentations alcohol was a factor. Thirty-eight of these patients attended within 24 hours of an Ireland match. The overall impact on the Emergency Department was limited. Introduction Ireland's participation in the Football World Cup Finals of 2002 led to intense media coverage and huge public interest. The Finals were held in South Korea and Japan. Many travelled to watch the Irish team but the vast majority of supporters had to be content with watching the televised coverage of the event. To illustrate this, on Irish terrestrial television alone, an estimated 1,714,000 people watched the televised Ireland v Spain match. This represents 53% of the population of Ireland. PMID- 12722788 TI - Current trends for infection prevention in total hip arthroplasty--a survey in Republic of Ireland. PMID- 12722790 TI - Immunohistochemical localization of androgen receptor in rat oocytes. AB - Immunohistochemical localization of androgen receptor (AR) was investigated in the rat oocytes during their development in the primary, secondary and antral follicles. The group of experimental Wistar rats included prepubertal female rats, killed at 30 days of age, and mature female rats killed at estrus or metestrus. Excised ovaries were submitted to immunohistochemical procedure in which polyclonal antibody against androgen receptor, avidin-biotin-peroxidase complex, and DAB were used. Characteristic changes in AR immunostaining intensity and localization in the oocyte compartments were observed during the oocyte growth. Relatively strong cytoplasmic AR immunostaining of oocytes from the primordial, primary and some secondary follicles became gradually weaker during the further oocyte development and was only weakly positive in the oocytes from the antral follicles. Germinal vesicles (GVs) usually displayed less intense AR immunostaining than cytoplasm and it was decreasing together with the cytoplasmic depletion of AR. On the contrary, nucleoli appeared as moderately AR-positive structures in the early secondary follicles and were strongly AR-positive in the multilaminar secondary and antral follicles. The presence of AR in the nucleoli persisted even when oocytes had undergone fragmentation in atretic follicles. These findings suggest that during the oocyte growth AR translocates from the oocyte cytoplasm to GV, and then to the nucleolus, which seems to become the main target for this receptor. A possible role of AR in the GV nucleolus is obscure. However, nucleolus contains rRNA genes and is the site of an active transcription, so the role of AR as a ligand-activated, transcriptional factor cannot be excluded. PMID- 12722791 TI - Immunohistochemical characterisation of cholinergic neurons in the anterior pelvic ganglion of the male pig. AB - This study investigated immunohistochemical properties of cholinergic neurons in the anterior pelvic ganglion (APG) of juvenile male pigs (n=7). Cholinergic neurons were identified using antibodies against choline acetyltransferase (ChAT) and vesicular acetylcholine transporter (VAChT). Immunoblotting was applied to verify the specificity of ChAT-immunostaining. Western blotting performed on APG tissue homogenates detected single immunoreactive protein with a molecular weight matching that of ChAT (71.6 kDa). It was found that many APG neurons expressed immunoreactivity to ChAT or VAChT (40% and 39% of the neurons, respectively). The analysis of adjacent sections from the ganglion revealed complete colocalization of ChAT and VAChT in these nerve cells. Furthermore, virtually all the ChAT positive neurons were tyrosine hydroxylase (TH)-negative (non-adrenergic) but many of them displayed immunoreactivity to nitric oxide synthase (NOS), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY) or somatostatin (SOM). There were also single nerve cell bodies that stained for neither ChAT nor TH. The comparison of the adjacent sections revealed that NOS, VIP, NPY and SOM were simultaneously co-expressed in the majority of the cholinergic somata. ChAT- or VAChT-positive varicose nerve terminals supplied nearly all neuronal profiles within the ganglion often forming loose basket-like formations surrounding the particular nerve cell bodies. The present study for the first time has revealed that nearly all non-adrenergic neurons in the porcine APG are cholinergic in nature, i.e. express immunoreactivity for ChAT and VAChT. Considering a high coincidence between the chemical coding of non-adrenergic (cholinergic) nerve fibres supplying some porcine male reproductive organs described in earlier papers and that of cholinergic pelvic neurons found in this study it is further concluded that pelvic ganglia are probably the major source of cholinergic innervation for the porcine urogenital system. PMID- 12722792 TI - Localization of neuropeptide Y and norepinephrine in the porcine ovarian artery and their influence on the local blood pressure. AB - The aim of the present study was to determine: (i) the presence of dopamine-beta hydroxylase (DbetaH)- and neuropeptide Y (NPY)-immunoreactive (IR) nerve fibres in the wall of the porcine ovarian artery, (ii) the influence of NPY and norepinephrine (NE) on the contractile activity of the pig ovarian arteries, and (iii) the pharmacological analysis of the interaction between NPY and NE in the isolated porcine ovarian arteries collected from immature pigs and from animals in different days of the estrous cycle. Ovarian arteries for immunohistochemistry and isolated arteries for pharmacological studies were excised from immature pigs and mature animals on days 1-5, 8-13 and 17-20 of the estrous cycle. The study showed that both DbetaH- and NPY-IR nerve fibres were present in the pig ovarian arteries in all periods examined, and, that in some fibres DbetaH and NPY were co localized. Both NE (10(-6) M) and NPY (10(-7) M) increased blood pressure of examined preparations, however, NE caused stronger changes in the vessel wall tension (P<0.001), than did NPY. NE significantly increased (P<0.001) blood pressure of all isolated arteries, however, this response was stronger in vessels from days 1-5 of the cycle, when compared to days 8-13 (P<0.01), 17-20 and immature pigs (P<0.001). NPY increased significantly blood pressure in isolated arteries from days 8-13 and 17-20 of the cycle (P<0.001), while in preparations taken from immature pigs and animals on days 1-5 of the estrous cycle this response was somewhat weaker (P<0.01). A higher elevation (P<0.001) of blood pressure after NPY administration was observed in isolated arteries from days 17 20 of the cycle, when compared to vessels from days 1-5 and 8-13 and those from immature pigs. Moreover, NE significantly intensified (P<0.001) an increase in the blood pressure in isolated arteries pre-treated with NPY in all periods examined. NPY insignificantly (P>0.05) potentiated increase of blood pressure in NE pre-treated vessels of immature pigs and in isolated arteries from days 17-20 of the cycle and significantly (P<0.05) in vessels from days 1-5 and 8-13 of the estrous cycle. Our results indicate that DbetaH- and NPY-IR nerve fibres are present in the pig ovarian arteries. NE and NPY administered alone increased blood pressure in the pig isolated ovarian artery and simultaneous administration of both substances caused each other potentiation of vasocontractile effect, however, the strength of observed changes was dependent on the stage of the estrous cycle. PMID- 12722793 TI - The evaluation of quantity and distribution of murine pulmonary mast cells in experimental hemorrhagic shock. AB - Respiratory failure is one of the most serious clinical complications in the course of post-hemorrhagic changes. Cascade-like, systemic inflammatory reaction including the axis: intestines-liver-lung-immune system has a special significance in the pathogenesis of this syndrome. In order to broaden the knowledge of the respiratory insufficiency pathogenesis in hemorrhagic shock, the attempt was made to evaluate quantitatively rat mast cells localized under the pulmonary pleura, around the bronchi, around large vessels and placed in the interalveolar septa. The examinations were conducted on 24 young female Wistar rats, divided into two groups (n=12): (I) sham-operated and (II) shocked. The hemorrhagic shock was evoked by the withdrawal of 25% of the circulating blood. The shock duration was 75 min. The obtained lung sections were stained with toluidine blue and examined in a light microscope. After hemorrhagic shock, sections of lung samples revealed about two-fold increase in mast cell number/mm2 compared to controls. Mast cells were concentrated mostly around the bronchi and blood vessels. Hyperplasia and migration of mast cells may suggest their role in the modulation of inflammatory process causing acute lung injury in the hemorrhagic shock. PMID- 12722794 TI - Immunohistochemical demonstration of nitric oxide synthase (NOS) in the normal rat pituitary gland, estrogen-induced rat pituitary tumor and human pituitary adenomas. AB - Nitric oxide synthase (NOS) immunoreactivity was examined in normal rat anterior pituitary glands, estrogen-induced rat pituitary tumors and human pituitary adenomas using a polyclonal antibody reacting with all three isoforms of NOS in both species. It was found that NOS immunorectivity in pituitary glandular cells is stronger in rat experimental pituitary tumors than in normal pituitaries. NOS immunoreactivity is also detectable in all but two human pituitary adenomas and seems to negatively correlate with microvascularization. PMID- 12722795 TI - Expression of C-ERBB2 and P65 genes and their protein products in follicular neoplasms of thyroid gland. AB - C-ERBB2 and P65 gene expression was investigated by reverse transcriptase polymerase chain reaction method (RT-PCR) in thirty follicular thyroid cancers and twenty follicular adenomas. Additionally, the cancers and adenomas were stained by immunohistochemistry for the expression of their protein products. We did not observe P65 gene expression in any of the analyzed follicular cancers (n=30) but it was observed in 13 out of 20 (65%) follicular adenomas. The presence of C-ERBB2 gene expression was found in 18 (90%) follicular adenomas but not in cancers. There were 10 (50%) adenomas and 11 (36.7%) cancers with positive staining for C-ERBB2 protein and 15 (75%) adenomas and 2 (6.7%) cancers with positive staining for P65 protein. We conclude that expression of C-ERBB2 and P65 genes is associated with follicular adenoma but not with cancer of thyroid gland. PMID- 12722796 TI - Evaluation of K-RAS gene in colorectal cancer. AB - The aim of the study was to assess the prevalence of K-RAS gene mutations in colorectal cancer and their role in diagnosis and prognosis. The study involved 36 patients with colorectal cancer at different stages of the disease progression and with different histopathologic grading. Mutations of codon 12 of K-RAS gene investigated using PCR-RFLP technique were found in 15 patients (41.67%). Although no statistically significant correlation was observed between the disease progression, histopathologic findings, gender and age, we suppose that assessment of K-RAS gene mutations might be of clinical value in the prognosis of colorectal cancer. PMID- 12722797 TI - Chromosomal aberrations in Crepis capillaris cells detected by FISH. AB - Crepis capillaris (2n=6) is an excellent plant for the assay of chromosome aberrations after mutagenic treatment. It has simple karyotype: three pairs of morphologically distinct and relatively large chromosomes. The frequency of structural chromosome aberrations and micronuclei in root meristem cells has been used for evaluation of the genotoxicity of chemicals and environmental pollutants. The introduction of fluorescence in situ hybridization method allows more detailed detection and localization of chromosomal rearrangements not only in mitotic but also in interphase nuclei. We demonstrate a few examples of the detection of chromosomal aberrations using rDNA and telomeric sequences as probes for in situ hybridization to C. capillaris chromosomes. PMID- 12722798 TI - Exposure to residential radon causes lung cancer. For the proposition. PMID- 12722799 TI - Exposure to residential radon causes lung cancer. Against the proposition. PMID- 12722800 TI - Are we doing any good by doing really well? (Where's the Bacon?). AB - Francis Bacon, who with Rene Decartes laid the intellectual foundations for Western science in the seventeenth century, asserted that the purpose of all knowledge is "action in the production of works for ... the relief of man's estate." We assess briefly several aspects of a few of the current efforts directed to the production of such "works" with respect to such "relief" as they may provide: cancer mortality, the medical literature, evidence-based medicine, clinical trials, observational databases and criteria for the promotion and tenure of the medical faculty. We suggest why each of these efforts appears to have failed to some degree and then propose some measures that may possibly serve as correctives. PMID- 12722801 TI - Thresholds for detecting and characterizing focal lesions using steady-state MR elastography. AB - An objective contrast-detail analysis was performed in this study to assess the low contrast detectability of a clinical prototype harmonic magnetic resonance elastographic imaging system. Elastographic imaging was performed on gelatin phantoms containing spherical inclusions of varying size and modulus contrast. The results demonstrate that lesions as small as 5 mm can be detected with a minimum modulus contrast of 14 dB. However, the shear modulus of such small lesions was not accurately recovered. In general, the shear modulus of larger focal lesions was accurately (i.e., within 25% of the true value) recovered. The minimum modulus contrast needed to detect focal lesions was observed to decrease with increasing lesion size. PMID- 12722802 TI - Quantifying the predictability of diaphragm motion during respiration with a noninvasive external marker. AB - The aim of this work was to quantify the ability to predict intrafraction diaphragm motion from an external respiration signal during a course of radiotherapy. The data obtained included diaphragm motion traces from 63 fluoroscopic lung procedures for 5 patients, acquired simultaneously with respiratory motion signals (an infrared camera-based system was used to track abdominal wall motion). During these sessions, the patients were asked to breathe either (i) without instruction, (ii) with audio prompting, or (iii) using visual feedback. A statistical general linear model was formulated to describe the relationship between the respiration signal and diaphragm motion over all sessions and for all breathing training types. The model parameters derived from the first session for each patient were then used to predict the diaphragm motion for subsequent sessions based on the respiration signal. Quantification of the difference between the predicted and actual motion during each session determined our ability to predict diaphragm motion during a course of radiotherapy. This measure of diaphragm motion was also used to estimate clinical target volume (CTV) to planning target volume (PTV) margins for conventional, gated, and proposed four-dimensional (4D) radiotherapy. Results from statistical analysis indicated a strong linear relationship between the respiration signal and diaphragm motion (p<0.001) over all sessions, irrespective of session number (p=0.98) and breathing training type (p=0.19). Using model parameters obtained from the first session, diaphragm motion was predicted in subsequent sessions to within 0.1 cm (1 sigma) for gated and 4D radiotherapy. Assuming a 0.4 cm setup error, superior-inferior CTV-PTV margins of 1.1 cm for conventional radiotherapy could be reduced to 0.8 cm for gated and 4D radiotherapy. The diaphragm motion is strongly correlated with the respiration signal obtained from the abdominal wall. This correlation can be used to predict diaphragm motion, based on the respiration signal, to within 0.1 cm (1 sigma) over a course of radiotherapy. PMID- 12722803 TI - Application of measured pencil beam parameters for electron beam model evaluation. AB - Most current electron beam models, as are used in commercial treatment planning systems, combine measured broad beam central axis depth dose data with measured or modeled functions to approximate radial scatter and heterogeneity effects. In this paper, we extend a recently developed pencil beam model to calculate doses outside the field edge and doses in heterogeneous media. We have also explored use of this model as a tool for evaluating commercial electron planning programs. The algorithm we have developed, based on the concept of the lateral buildup ratio (LBR), enables calculation of dose at any point in an irregular electron field, and is capable of generating both on- and off-axis depth dose curves and isodose profiles. This model includes the effects of density and mass-angular scattering power in measured broad beam central axis depth dose data, which when combined with small field reference data, can be used to generate LBR ratios. From these ratios one can infer the depth dependent, effective pencil beam radial spread parameter a in water or other materials, which can be used to model any arbitrary field. We have used this approach to calculate fractional depth doses for small fields incident on aluminum and cork, which we have then compared against measurements and the calculations of several commercial planning systems. PMID- 12722805 TI - Head scatter off-axis for megavoltage x rays. AB - Our objective in this study has been to investigate how head scatter varies with the off-axis position in a 6 MV x-ray beam. We define the head-scatter off-axis ratio, HOA, as the ratio of the kerma due to head-scatter photons at the off-axis position x to the kerma from direct primary photons on the central axis. "Direct primary" are those photons that come from the source without interactions in the intervening structures. We determined HOA from measurements with an ionization chamber in a miniphantom. Head-scatter and direct primary photons contribute to a measurement of the ionization per mu Q(x) at the off-axis position x in the open field cx x cy. The ionization per mu QP(x), measured in the same position but with the field collimated to the smallest possible opening (cx x 3 cm), is intended to include only direct primary photons. Head-scatter photons cannot be completely eliminated, and the errors due to remaining head scatter and radiation back-scattered by the movable collimators into the monitor were estimated. For normalization of the final results, ionization due to direct primary photons was also measured on the central axis, QP(0). HOA was derived from these three measurements as HOA(cx,cy,x)=(Q(cx,cy,x) - QP(cx,cy,x))/QP(cx,cy,0). On the central axis (x=y=0), HOA represents the "scatter-to-primary ratio" between head scatter and the direct primary dose. Monte Carlo simulations were made to help with the interpretation and evaluation of the results. HOA could be fitted to a Gaussian model with two components corresponding to sources of widths 1.8 and 14 cm, projected on a plane 5 cm below the x-ray source. The narrow Gaussian component is interpreted as the source of photons scattered in the flattening filter and the primary collimator. The broad component is attributed to photons scattered in the secondary (variable) collimators. Conventional head-scatter models (e.g., a single Gaussian source model) do not fit the measured HOA data for large collimator settings (c>20 cm) or outside beam collimation. The full width at half-maximum (FWHM) of HOA(x) across the field increased with the field width (cx) in the direction of the measurements in a manner consistent with the field of view of the two sources. It was not sensitive to the field measure in the orthogonal direction (cy). Head scatter outside the field also increased with field size, reflecting an increased contribution of photons scattered at large angles. It exceeds the leakage through the collimator 2 cm outside the edge for square fields c>10 cm. Monte Carlo calculations showed considerably less head scatter outside the field than measurements. PMID- 12722804 TI - Monte Carlo calculated correction factors for primary standards of air kerma. AB - Many laboratories with cavity chambers as primary standards for air kerma are considering using additional Monte Carlo calculated correction factors, in particular the correction for attenuation and scatter in the walls, Kwall, and possibly the correction for point of measurement, Kan. Standards labs also use Monte Carlo calculated Spencer-Attix stopping-power ratios for graphite to air. The purpose of this article is to investigate the sensitivity of these calculations to their details and to assign uncertainties to the calculated values. We also investigate the correction needed for the Canadian primary standard to account for a polystyrene insulator, Kcomp and find that it is quite large (1.0046 +/- 0.0017). The article shows that the values of correction factors are very robust and insensitive to most details of the calculations except the values of the underlying electron stopping powers which have a significant effect on the stopping-power ratio and on Kcomp. The 1% uncertainties on the photon cross-sections have a negligible effect on these correction factors except for Kcomp. As a result of these investigations, with no change in the stopping power data used, the Canadian primary standard of air kerma in a 60Co beam needs to be increased by 0.54%. PMID- 12722806 TI - IMRT dose shaping with regionally variable penalty scheme. AB - A commonly known deficiency of currently available inverse planning systems is the difficulty in fine-tuning the final dose distribution. In practice, it is not uncommon that just a few unsatisfactory regions in the planning target volume or an organ at risk prevent an intensity modulated radiation therapy (IMRT) plan from being clinically acceptable. The purpose of this work is to introduce a mechanism for controlling the regional doses after a conventional IMRT plan is obtained and to demonstrate its clinical utility. Two types of importance factors are introduced in the objective function to model the tradeoffs of different clinical objectives. The first is the conventional structure-dependent importance factor, which quantifies the interstructure tradeoff. The second type is the voxel-dependent importance factor which "modulates" the importance of different voxels within a structure. The planning proceeds in two major steps. First a conventional inverse planning is performed, where the structure-dependent importance factors are determined in a trial-and-error fashion. The next level of planning involves fine-tuning the regional doses to meet specific clinical requirements. To achieve this, the voxels where doses need to be modified are identified either graphically on the isodose layouts, or on the corresponding dose-volume histogram (DVH) curves. The importance value of these voxels is then adjusted to increase/decrease the penalty at the corresponding regions. The technique is applied to two clinical cases. It was found that both tumor hot spots and critical structure maximal doses can be easily controlled by varying the regional penalty. One to three trials were sufficient for the conventionally optimized dose distributions to be adjusted to meet clinical expectation. Thus introducing the voxel-dependent penalty scheme provides an effective means for IMRT dose distributions painting and sculpting. PMID- 12722808 TI - Photon beam relative dose validation of the DPM Monte Carlo code in lung equivalent media. AB - Validation experiments have been conducted using 6 and 15 MV photons in inhomogeneous (water/lung/water) media to benchmark the accuracy of the DPM Monte Carlo code for photon beam dose calculations. Small field sizes (down to 2 x 2 cm2) and low-density media were chosen for this investigation because the intent was to test the DPM code under conditions where lateral electronic disequilibrium effects are emphasized. The treatment head components of a Varian 21EX linear accelerator, including the jaws (defining field sizes of 2 x 2, 3 x 3 and 10 x 10 cm2), were simulated using the BEAMnrc code. The phase space files were integrated within the DPM code system, and central axis depth dose and profile calculations were compared against diode measurements in a homogeneous water phantom in order to validate the phase space. Results of the homogeneous phantom study indicated that the relative differences between DPM calculations and measurements were within +/- 1% (based on the rms deviation) for the depth dose curves; relative profile dose differences were on average within +/- 1%/1 mm. Depth dose and profile measurements were carried out using an ion-chamber and film, within an inhomogeneous phantom consisting of a 6 cm slab of lung equivalent material embedded within solid water. For the inhomogeneous phantom experiment, DPM depth dose calculations were within +/- 1% (based on the rms deviation) of measurements; relative profile differences at depths within and beyond the lung were, on average, within +/- 2% in the inner and outer beam regions, and within 1-2 mm distance-to-agreement within the penumbral region. Relative point differences on the order of 2-3% were within the estimated experimental uncertainties. This work demonstrates that the DPM Monte Carlo code is capable of accurate photon beam dose calculations in situations where lateral electron disequilibrium effects are pronounced. PMID- 12722807 TI - Quantifying the effect of intrafraction motion during breast IMRT planning and dose delivery. AB - Respiratory motion during intensity modulated radiation therapy (IMRT) causes two types of problems. First, the clinical target volume (CTV) to planning target volume (PTV) margin needed to account for respiratory motion means that the lung and heart dose is higher than would occur in the absence of such motion. Second, because respiratory motion is not synchronized with multileaf collimator (MLC) motion, the delivered dose is not the same as the planned dose. The aims of this work were to evaluate these problems to determine (a) the effects of respiratory motion and setup error during breast IMRT treatment planning, (b) the effects of the interplay between respiratory motion and multileaf collimator (MLC) motion during breast IMRT delivery, and (c) the potential benefits of breast IMRT using breath-hold, respiratory gated, and 4D techniques. Seven early stage breast cancer patient data sets were planned for IMRT delivered with a dynamic MLC (DMLC). For each patient case, eight IMRT plans with varying respiratory motion magnitudes and setup errors (and hence CTV to PTV margins) were created. The effects of respiratory motion and setup error on the treatment plan were determined by comparing the eight dose distributions. For each fraction of these plans, the effect of the interplay between respiratory motion and MLC motion during IMRT delivery was simulated by superimposing the respiratory trace on the planned DMLC leaf motion, facilitating comparisons between the planned and expected dose distributions. When considering respiratory motion in the CTV-PTV expansion during breast IMRT planning, our results show that PTV dose heterogeneity increases with respiratory motion. Lung and heart doses also increase with respiratory motion. Due to the interplay between respiratory motion and MLC motion during IMRT delivery, the planned and expected dose distributions differ. This difference increases with respiratory motion. The expected dose varies from fraction to fraction. However, for the seven patients studied and respiratory trace used, for no breathing, shallow breathing, and normal breathing, there were no statistically significant differences between the planned and expected dose distributions. Thus, for breast IMRT, intrafraction motion degrades treatment plans predominantly by the necessary addition of a larger CTV to PTV margin than would be required in the absence of such motion. This motion can be limited by breath-hold, respiratory gated, or 4D techniques. PMID- 12722809 TI - Determining the incident electron fluence for Monte Carlo-based photon treatment planning using a standard measured data set. AB - An accurate dose calculation in phantom and patient geometries requires an accurate description of the radiation source. Errors in the radiation source description are propagated through the dose calculation. With the emergence of linear accelerators whose dosimetric characteristics are similar to within measurement uncertainty, the same radiation source description can be used as the input to dose calculation for treatment planning at many institutions with the same linear accelerator model. Our goal in the current research was to determine the initial electron fluence above the linear accelerator target for such an accelerator to allow a dose calculation in water to within 1% or 1 mm of the measured data supplied by the manufacturer. The method used for both the radiation source description and the patient transport was Monte Carlo. The linac geometry was input into the Monte Carlo code using the accelerator's manufacturer's specifications. Assumptions about the initial electron source above the target were made based on previous studies. The free parameters derived for the calculations were the mean energy and radial Gaussian width of the initial electron fluence and the target density. A combination of the free parameters yielded an initial electron fluence that, when transported through the linear accelerator and into the phantom, allowed a dose-calculation agreement to the experimental ion chamber data to within the specified criteria at both 6 and 18 MV nominal beam energies, except near the surface, particularly for the 18 MV beam. To save time during Monte Carlo treatment planning, the initial electron fluence was transported through part of the treatment head to a plane between the monitor chambers and the jaws and saved as phase-space files. These files are used for clinical Monte Carlo-based treatment planning and are freely available from the authors. PMID- 12722810 TI - MOSFET dosimetry for microbeam radiation therapy at the European Synchrotron Radiation Facility. AB - Preclinical experiments are carried out with approximately 20-30 microm wide, approximately 10 mm high parallel microbeams of hard, broad-"white"-spectrum x rays (approximately 50-600 keV) to investigate microbeam radiation therapy (MRT) of brain tumors in infants for whom other kinds of radiotherapy are inadequate and/or unsafe. Novel physical microdosimetry (implemented with MOSFET chips in the "edge-on" mode) and Monte Carlo computer-simulated dosimetry are described here for selected points in the peak and valley regions of a microbeam-irradiated tissue-equivalent phantom. Such microbeam irradiation causes minimal damage to normal tissues, possible because of rapid repair of their microscopic lesions. Radiation damage from an array of parallel microbeams tends to correlate with the range of peak-valley dose ratios (PVDR). This paper summarizes comparisons of our dosimetric MOSFET measurements with Monte Carlo calculations. Peak doses at depths <22 mm are 18% less than Monte Carlo values, whereas those depths >22 mm and valley doses at all depths investigated (2 mm-62 mm) are within 2-13% of the Monte Carlo values. These results lend credence to the use of MOSFET detector systems in edge-on mode for microplanar irradiation dosimetry. PMID- 12722811 TI - Image reconstruction with shift-variant filtration and its implication for noise and resolution properties in fan-beam computed tomography. AB - In computed tomography (CT), the fan-beam filtered backprojection (FFBP) algorithm is used widely for image reconstruction. It is known that the FFBP algorithm can significantly amplify data noise and aliasing artifacts in situations where the focal lengths are comparable to or smaller than the size of the field of measurement (FOM). In this work, we propose an algorithm that is less susceptible to data noise, aliasing, and other data inconsistencies than is the FFBP algorithm while retaining the favorable resolution properties of the FFBP algorithm. In an attempt to evaluate the noise properties in reconstructed images, we derive analytic expressions for image variances obtained by use of the FFBP algorithm and the proposed algorithm. Computer simulation studies are conducted for quantitative evaluation of the spatial resolution and noise properties of images reconstructed by use of the algorithms. Numerical results of these studies confirm the favorable spatial resolution and noise properties of the proposed algorithm and verify the validity of the theoretically predicted image variances. The proposed algorithm and the derived analytic expressions for image variances can have practical implications for both estimation and detection/classification tasks making use of CT images, and they can readily be generalized to other fan-beam geometries. PMID- 12722813 TI - An experimental comparison of detector performance for direct and indirect digital radiography systems. AB - Current flat-panel detectors either directly convert x-ray energy to electronic charge or use indirect conversion with an intermediate optical process. The purpose of this work was to compare direct and indirect detectors in terms of their modulation transfer function (MTF), noise power spectrum (NPS), and detective quantum efficiency (DQE). Measurements were made on three flat-panel detectors, Hologic Direct-Ray DR-1000 (DRC), GE Revolution XQ/i (XQ/i), and Philips Digital Diagnost (DiDi) using the IEC-defined RQA5 (approximately 74 kVp, 21 mm Al) and RQA9 (approximately 120 kVp, 40 mm Al) radiographic techniques. The presampled MTFs of the systems were measured using an edge method [Samei et al., Med. Phys. 25, 102 (1998)]. The NPS of the systems were determined for a range of exposure levels by two-dimensional (2D) Fourier analysis of uniformly exposed radiographs [Flynn and Samei, Med. Phys. 26, 1612 (1999)]. The DQEs were assessed from the measured MTF, NPS, exposure, and estimated ideal signal-to-noise ratios. For the direct system, the MTF was found to be significantly higher than that for the indirect systems and very close to an ideal function associated with the detector pixel size. The NPS for the direct system was found to be constant in relation to frequency. For the XQ/i and DRC systems, the DQE results reflected expected differences based on the absorption efficiency of the different detector materials. Using RQA5, the measured DQE values in the diagonal (and axial) direction(s) at spatial frequencies of 0.15 mm(-1) and 2.5 mm(-1) were 64% (64%) and 20% (15%) for the XQ/i system, and 38% (38%) and 20% (20%) for the DRC, respectively. The DQE results of the DiDi system were difficult to interpret due to additional preprocessing steps in that system. PMID- 12722812 TI - Evaluation of a flat panel digital radiographic system for low-dose portable imaging of neonates. AB - The purpose of this study was to evaluate the clinical utility of an investigational flat-panel digital radiography system for low-dose portable neonatal imaging. Thirty image-pairs from neonatal intensive care unit patients were acquired with a commercial Computed Radiography system (Agfa, ADC 70), and with the investigational system (Varian, Paxscan 2520) at one-quarter of the exposure. The images were evaluated for conspicuity and localization of the endings of ancillary catheters and tubes in two observer performance experiments with three pediatric radiologists and three neonatologists serving as observers. The results indicated no statistically significant difference in diagnostic quality between the images from the investigational system and from CR. Given the investigational system's superior resolution and noise characteristics, observer results suggest that the high detective quantum efficiency of flat-panel digital radiography systems can be utilized to decrease the radiation dose/exposure to neonatal patients, although post-processing of the images remains to be optimized. The rapid availability of flat-panel images in portable imaging was found to be an added advantage for timely clinical decision-making. PMID- 12722814 TI - Optical-CT gel-dosimetry I: basic investigations. AB - Comprehensive verification of the intricate dose distributions associated with advanced radiation treatments is now an immediate and substantial problem. The task is challenging using traditional dosimeters because of restrictions to point measurements (ion chambers, diodes, TLD, etc.) or planar measurements (film). In essence, rapid advances in the technology to deliver radiation treatments have not been paralleled by corresponding advances in the ability to verify these treatments. A potential solution has emerged in the form of water equivalent three dimensional (3D) gel-dosimetry. In this paper we present basic characterization and performance studies of a prototype optical-CT scanning system developed in our laboratory. An analysis of the potential role or scope of gel dosimetry, in relation to other dosimeters, and to verification across the spectrum of therapeutic techniques is also given. The characterization studies enabled the determination of nominal operating conditions for optical-CT scanning. "Finger" phantoms are introduced as a powerful and flexible tool for the investigation of optical-CT performance. The modulation-transfer function (MTF) of the system is determined to be better than 10% out to 1 mm(-1), confirming sub-mm imaging ability. System performance is demonstrated by the acquisition of a 1 x 1 x 1 mm3 dataset through the dose distribution delivered by an x-ray lens that focuses x rays in the energy range 40-80 KeV. This 3D measurement would be extremely difficult to achieve with other dosimetry techniques and highlights some of the strengths of gel dosimetry. Finally, an optical Monte Carlo model is introduced and shown to have potential to model light transport through gel-dosimetry systems, and to provide a tool for the study and optimization of optical-CT gel dosimetry. The model utilizes Mie scattering theory and requires knowledge of the variation of the particle size distribution with dose. The latter was determined here using the technique of dynamic-light-scattering. PMID- 12722815 TI - Review of the energy check of an electron-only linear accelerator over a 6 year period: sensitivity of the technique to energy shift. AB - The calibration and monthly QA of an electron-only linear accelerator dedicated to intra-operative radiation therapy has been reviewed. Since this machine is calibrated prior to every procedure, there was no necessity to adjust the output calibration at any time except, perhaps, when the magnetron is changed, provided the machine output is reasonably stable. This gives a unique opportunity to study the dose output of the machine per monitor unit, variation in the timer error, flatness and symmetry of the beam and the energy check as a function of time. The results show that, although the dose per monitor unit varied within +/- 2%, the timer error within +/- 0.005 MU and the asymmetry within 1-2%, none of these parameters showed any systematic change with time. On the other hand, the energy check showed a linear drift with time for 6, 9, and 12 MeV (2.1, 3.5, and 2.5%, respectively, over 5 years), while at 15 and 18 MeV, the energy check was relatively constant. It is further shown that based on annual calibrations and RPC TLD checks, the energy of each beam is constant and that therefore the energy check is an exquisitely sensitive one. The consistency of the independent checks is demonstrated. PMID- 12722816 TI - Temporal resolution and the evaluation of candidate algorithms for four dimensional CT. AB - The four-dimensional computed tomography ("4D-CT") with area detector has been developed for dynamic volumetric imaging with large longitudinal coverage. In this paper one of the key technologies for 4D-CT development is discussed: Image reconstruction algorithm with high temporal resolution. All of the cone-beam algorithms investigated previously assume that the object is stationary. In this paper a new class of cone-beam problem is addressed: a dynamic volumetric (4-D) imaging. A continuously rotating circular (stationary couch) scanning is employed, and then, a generalized version of the well-known Feldkamp algorithm with the following three steps is performed: (1) applying a weighting function (along the time axis) to projection data, (2) filtering the weighted data along the detector row direction, (3) cone-beam backprojecting of the filtered data along the corresponding x-ray path. The weighting function controls the time center, the temporal resolution, and the image quality. Four weighting functions developed for fan-beam reconstruction were applied to the first step: (a) a constant weight fixed at 0.5 (FS-FDK), (b) feathering both edges of the (time) window (OS-FDK), (c) Parker's weight for a half-scan (HF-FDK), and (d) an extended Parker's weight, which allows us to use a larger range of projection data up to one rotation (NHS-FDK). We evaluated them in terms of temporal resolution, image noise, and image quality. Also, the cause of the artifact has been investigated. The temporal resolution of NHF-FDK equals that of HS-FDK, which is half of the one rotation period. For the moving object, NHS-FDK offers the best image quality. The images with FS-FDK are degraded by streak artifacts; HS-FDK provides poor image quality with good temporal resolution; and images by OS-FDK are blurred due to insufficient temporal resolution. The cause of the artifact was found as an inconsistency of projection data due to object motion (in FS-FDK) and lost 3-D-Radon data caused by applying Parker's weight (in HS FDK). A hand toy was employed for the preliminary evaluation of dynamic volumetric imaging with the real 256-slice scanner. In an overall evaluation, NHS FDK provides the stable and the sufficient image quality both with moving and stationary objects. PMID- 12722817 TI - Technical note: fitted dosimetric parameters of high dose-rate 192Ir sources according to the AAPM TG43 formalism. AB - Functional fits for the anisotrophy function and the radial dose function, have been studied, in a previous work, in order to characterize dose-rate distributions around some of the high-intensity 192Ir sources. The purpose of the present work is to complete the previous one in order to include all the existing HDR and PDR 192Ir sources. The sources addressed here are: the Buchler source from Amersham, the 12i and Plus PDR sources and the 12i and Plus HDR sources from GammaMed, and the new VariSource HDR source wire model VS2000 from Varian Oncology Systems. PMID- 12722818 TI - Therapeutic treatment plan optimization with probability density-based dose prescription. AB - The dose optimization in inverse planning is realized under the guidance of an objective function. The prescription doses in a conventional approach are usually rigid values, defining in most instances an ill-conditioned optimization problem. In this work, we propose a more general dose optimization scheme based on a statistical formalism [Xing et al., Med. Phys. 21, 2348-2358 (1999)]. Instead of a rigid dose, the prescription to a structure is specified by a preference function, which describes the user's preference over other doses in case the most desired dose is not attainable. The variation range of the prescription dose and the shape of the preference function are predesigned by the user based on prior clinical experience. Consequently, during the iterative optimization process, the prescription dose is allowed to deviate, with a certain preference level, from the most desired dose. By not restricting the prescription dose to a fixed value, the optimization problem becomes less ill-defined. The conventional inverse planning algorithm represents a special case of the new formalism. An iterative dose optimization algorithm is used to optimize the system. The performance of the proposed technique is systematically studied using a hypothetical C-shaped tumor with an abutting circular critical structure and a prostate case. It is shown that the final dose distribution can be manipulated flexibly by tuning the shape of the preference function and that using a preference function can lead to optimized dose distributions in accordance with the planner's specification. The proposed framework offers an effective mechanism to formalize the planner's priorities over different possible clinical scenarios and incorporate them into dose optimization. The enhanced control over the final plan may greatly facilitate the IMRT treatment planning process. PMID- 12722819 TI - A video analysis technique for organ dose assessment in pediatric fluoroscopy: applications to voiding cystourethrograms (VCUG). AB - The time-sequence videotape-analysis methodology, originally developed by Sulieman et al. [Radiol. 178, 653-658 (1991)] for use in tissue dose estimation in adult fluoroscopy exams, has been adapted to the study of the newborn voiding cystourethrogram (VCUG). Individual frames of fluoroscopic and radiographic video were analyzed with respect to unique combinations of field size, field center, projection, tube potential, and mA or mAs, respectively. A modified version of the stylized ORNL newborn model was coupled to the MCNP4C radiation transport code to report organ doses per unit entrance air kerma (free-in-air) for each identified x-ray field. A series of urinary bladder models was additionally developed representing the organ at differing stages of contrast filling. The technique was subsequently applied to two patients, a 3-month male and a 1-month female, examined via a conventional fluoroscopy system used just prior to departmental conversion to digital systems. The effective dose to these patients was estimated as 0.47 mSv and 1.36 mSv, respectively (ratio of 2.9). Corresponding ratios of cumulative fluoroscopy time and entrance air kerma were 2.2 and 1.6, respectively. For the male patient, the mean percent dose contribution from fluoroscopy for all irradiated organs was 71 +/- 12%, while that value for the female patient was 88 +/- 4%. PMID- 12722820 TI - Vessel size measurements in angiograms: manual measurements. AB - Vessel size measurement is perhaps the most often performed quantitative analysis in diagnostic and interventional angiography. Although automated vessel sizing techniques are generally considered to have good accuracy and precision, we have observed that clinicians rarely use these techniques in standard clinical practice, choosing to indicate the edges of vessels and catheters to determine sizes and calibrate magnifications, i.e., manual measurements. Thus, we undertook an investigation of the accuracy and precision of vessel sizes calculated from manually indicated edges of vessels. Manual measurements were performed by three neuroradiologists and three physicists. Vessel sizes ranged from 0.1-3.0 mm in simulation studies and 0.3-6.4 mm in phantom studies. Simulation resolution functions had full-widths-at-half-maximum (FWHM) ranging from 0.0 to 0.5 mm. Phantom studies were performed with 4.5 in., 6 in., 9 in., and 12 in. image intensifier modes, magnification factor = 1, with and without zooming. The accuracy and reproducibility of the measurements ranged from 0.1 to 0.2 mm, depending on vessel size, resolution, and pixel size, and zoom. These results indicate that manual measurements may have accuracies comparable to automated techniques for vessels with sizes greater than 1 mm, but that automated techniques which take into account the resolution function should be used for vessels with sizes smaller than 1 mm. PMID- 12722821 TI - A Grangeat-type half-scan algorithm for cone-beam CT. AB - Modern CT and micro-CT scanners are rapidly moving from fan-beam toward cone-beam geometry. Half-scan CT algorithms are advantageous in terms of temporal resolution, and widely used in fan-beam and cone-beam geometry. While existing half-scan algorithms for cone-beam CT are in the Feldkamp framework, in this paper we compensate missing data explicitly in the Grangeat framework, and formulate a half-scan algorithm in the circular scanning case. The half-scan spans 180 degrees plus two cone angles that guarantee sufficient data for reconstruction of the midplane defined by the source trajectory. The smooth half scan weighting functions are designed for the suppression of data inconsistency. Numerical simulation results are reported for verification of our formulas and programs. This Grangeat-type half-scan algorithm produces excellent image quality, without off-mid-plane artifacts associated with Feldkamp-type half-scan algorithms. The Grangeat-type half-scan algorithm seems promising for quantitative and dynamic biomedical applications of CT and micro-CT. PMID- 12722822 TI - Brachytherapy dosimetry of 125I and 103Pd sources using an updated cross section library for the MCNP Monte Carlo transport code. AB - Permanent implantation of low energy (20-40 keV) photon emitting radioactive seeds to treat prostate cancer is an important treatment option for patients. In order to produce accurate implant brachytherapy treatment plans, the dosimetry of a single source must be well characterized. Monte Carlo based transport calculations can be used for source characterization, but must have up to date cross section libraries to produce accurate dosimetry results. This work benchmarks the MCNP code and its photon cross section library for low energy photon brachytherapy applications. In particular, we calculate the emitted photon spectrum, air kerma, depth dose in water, and radial dose function for both 125I and 103Pd based seeds and compare to other published results. Our results show that MCNP's cross section library differs from recent data primarily in the photoelectric cross section for low energies and low atomic number materials. In water, differences as large as 10% in the photoelectric cross section and 6% in the total cross section occur at 125I and 103Pd photon energies. This leads to differences in the dose rate constant of 3% and 5%, and differences as large as 18% and 20% in the radial dose function for the 125I and 103Pd based seeds, respectively. Using a partially updated photon library, calculations of the dose rate constant and radial dose function agree with other published results. Further, the use of the updated photon library allows us to verify air kerma and depth dose in water calculations performed using MCNP's perturbation feature to simulate updated cross sections. We conclude that in order to most effectively use MCNP for low energy photon brachytherapy applications, we must update its cross section library. Following this update, the MCNP code system will be a very effective tool for low energy photon brachytherapy dosimetry applications. PMID- 12722823 TI - Monte Carlo dosimetry of 60Co HDR brachytherapy sources. AB - Monte Carlo based dosimetry is presented for three 60Co HDR sources with a unique configuration of two active pellets in contact or spaced 9 and 11 mm apart. Results are presented in Cartesian "away and along" as well as polar coordinates following the AAPM TG-43 dosimetric formalism. Iso-dose rate contours around the sources in Cartesian coordinates reveal that significant differences between the three source designs exist only close to the source centers where dose rate distributions bear the effect of the unique source configurations. Dose rate constants of all three sources are accurately described by an equation of the form: lambda(cGyh(-1)U(-1))=lambda*pointG(1 cm,90 degrees) = 1.094*G(1 cm,90 degrees) where lambdapoint is the dose rate constant of a bare 60Co point source and G(1 cm,90 degrees) is the "exact" source geometry factor as defined by the TG 43. Radial dose and anisotropy function data extracted using the point source approximated geometry factors are tabulated for all three source designs. Finally, the dependence and variation of the above TG-43 parameters are discussed and it is demonstrated that the dosimetric properties of high-energy photon emitters are largely dependent on radionuclide source distribution. PMID- 12722824 TI - Comment on "Analytical representation of enhanced dynamic wedge factors for symmetric and asymmetric fields". PMID- 12722826 TI - Neurotrophins and neurodegenerative diseases: receptors stuck in traffic? AB - Neurotrophins are well known for their physiological role as key modulators of neuronal survival, neurite out-growth, and synaptic connectivity during development and into adulthood. Moreover, neurotrophins are potent agents, ameliorating neuronal degeneration in many model systems for neurological diseases. However, a causal role for mutations in neurotrophins or neurotrophin receptors in human neurodegenerative diseases has been largely lacking. As neurotrophin receptors are located at synapses and as their signaling involves the neuronal nucleus, they need to bridge tantalizing distances in order to retrogradely communicate their survival signals. On the other hand, anterogradely transported neurotrophins are released at the synapse and act on postsynaptic cells. Antero- and retrograde signaling and trafficking is an emerging focus of interest in neurotrophin research. Some neurodegenerative diseases are known to affect transport of organelles. Thus, it appears likely that neurodegeneration could be caused by "indirect" effects on neurotrophin trafficking and, hence, signaling. In this review we summarize recent work on neurotrophins in neurodegenerative diseases with special focus on possible implications of disturbed trafficking of organelles and retrograde axonal signaling. PMID- 12722825 TI - Signaling of cell death and cell survival following focal cerebral ischemia: life and death struggle in the penumbra. AB - Focal ischemia by middle cerebral artery occlusion (MCAO) results in necrosis at the infarct core and activation of complex signal pathways for cell death and cell survival in the penumbra. Recent studies have shown activation of the extrinsic and intrinsic pathways of caspase-mediated cell death, as well as activation of the caspase-independent signaling pathway of apoptosis in several paradigms of focal cerebral ischemia by transient MCAO to adult rats and mice. The extrinsic pathway (cell-death receptor pathway) is initiated by activation of the Fas receptor after binding to the Fas ligand (Fas-L); increased Fas and Fas-L expression has been shown following focal ischemia. Moreover, focal ischemia is greatly reduced in mice expressing mutated (nonfunctional) Fas. Increased expression of caspase-1, -3, -8, and -9, and of cleaved caspase-8, has been observed in the penumbra. Activation of the intrinsic (mitochondrial) pathway following focal ischemia is triggered by Bax translocation to and competition with Bcl-2 and other members of the Bcl-2 family in the mitochondria membrane that is followed by cytochrome c release to the cytosol. Bcl-2 over-expression reduces infarct size. Cytochrome c binds to Apaf-1 and dATP and recruits and cleaves pro-caspase-9 in the apoptosome. Both caspase-8 and caspase-9 activate caspase-3, among other caspases, which in turn cleave several crucial substrates, including the DNA-repairing enzyme poly(ADP-ribose) polymerase (PARP), into fragments of 89 and 28 kDa. Inhibition of caspase-3 reduces the infarct size, further supporting caspase-3 activation following transient MCAO. In addition, caspase-8 cleaves Bid, the truncated form of which has the capacity to translocate to the mitochondria and induce cytochrome c release. The volume of brain infarct is greatly reduced in Bid-deficient mice, thus indicating activation of the mitochondrial pathway by cell-death receptors following focal ischemia. Recent studies have shown the mitochondrial release of other factors; Smac/DIABLO (Smac: second mitochondrial activator of caspases: DIABLO: direct IAP binding protein with low pI) binds to and neutralizes the effects of the X-linked inhibitor of apoptosis (XIAP). Finally, apoptosis-inducing factor (AIF) translocates to the mitochondria and the nucleus following focal ischemia and produces peripheral chromatin condensation and large-scale DNA strands, thus leading to the caspase-independent cell death pathway of apoptosis. Delineation of the pro-apoptotic and pro-survival signals in the penumbra may not only increase understanding of the process but also help to rationalize strategies geared to reducing brain damage targeted at the periphery of the infarct core. PMID- 12722827 TI - Neuron-conditioned media differentially affect the survival of activated or unstimulated microglia: evidence for neuronal control on apoptotic elimination of activated microglia. AB - It is presently unknown what types of neuronal signals maintain microglial cells resting in the normal brain or control their activation in neuropathology. Recent data suggest that microglia activation induces apoptosis and that healthy neurons are controllers of the activation state and immune functions of microglia. In the present study we have evaluated, on microglial cells in cultures, whether neurons are able to affect their survival in resting conditions or upon activation with the bacterial endotoxin, lipopolysaccharide (LPS). We report that neuron conditioned culture media induced apoptosis of LPS-stimulated, but not of unstimulated, microglia. This effect was, however, only present when conditioned media had been exposed to differentiated neurons and not to immature ones, and was absent when glutamate receptors had been pharmacologically blocked in neuronal cultures. The effect was also blocked by heat-inactivation of the conditioned media. Media conditioned with either differentiated or undifferentiated cerebellar granule neurons positively affected the survival of unstimulated microglial cells when the standard concentration of fetal bovine serum (10%) was included in the culture media. Our results highlight the ability of differentiated neurons to maintain a controlled inflammatory state through production of factor(s) favoring the apoptotic elimination of activated microglia. They also suggest that immature neurons may, on the contrary, favor the survival of microglia during development. PMID- 12722828 TI - Regional distribution of alpha-synuclein pathology in unimpaired aging and Alzheimer disease. AB - The amygdaloid complex (AC) was found highly vulnerable to alpha-synuclein (alphaS) pathology in both familial and sporadic Alzheimer disease (AD), and recently, incidental Lewy bodies (LBs) were identified primarily in the lower brainstem. This challenges the traditional view that the substantia nigra (SN) is the region that is predominately affected in the spectrum of LB disorders. We examined the immunoreactivity of alphaS in the SN, the nucleus basalis of Meynert (nbM), and the AC in 904 subjects with or without concomitant AD pathology. AlphaS-positive structures were seen in at least one of the studied brain areas in 121 subjects (13%). The affected regions in the alphaS-positive subjects included the SN (89%), the nbM (73%), and the AC (67%). This study also included 82 sporadic AD patients diagnosed using CERAD criteria. AlphaS-positive structures were seen in 32% of the AD patients, with the SN and AC being equally affected. In a few subjects the AC was the only affected area. However, this was not inevitably associated with AD pathology, but was related to cognitive decline. Incidental LBs in the SN were described in the occasional subjects, with no alphaS pathology in the lower brainstem. PMID- 12722830 TI - Hippocampal and entorhinal cortex neurofibrillary tangle formation in Guamanian Chamorros free of overt neurologic dysfunction. AB - Since first described, amyotrophic lateral sclerosis/parkinsonism-dementia complex (ALS/PDC) of Guam has represented an important model of age-related neurodegenerative disease. ALS/PDC is characterized neuropathologically by severe widespread involvement by neurofibrillary tangles (NFTs). Over the past 30 years there has been a dramatic decrease in the incidence of ALS and a 10-year increase in the age of onset of ALS and PDC. In 1979, Anderson et al reported evidence of significant NFT involvement in Guam natives who had been free of evidence of neurologic dysfunction. Using the slides from this study, we re-examined the extent of hippocampus and entorhinal NFT involvement and compared it to brains recently obtained from neurologically intact Guam natives and age-matched controls from New York. The tendency towards hippocampal and entorhinal NFT formation continues to be encountered among the inhabitants of Guam, particularly among those over age 50. although severe involvement was less commonly noted in relatively young individuals (< 50 years). As noted by Anderson et al, the pattern of neuropathologic lesions seen in those with extensive NFT involvement suggests that such cases represent preclinical examples of ALS/PDC in individuals who have yet to accumulate a sufficient burden of pathology to attract clinical attention and diagnostic evaluation. PMID- 12722829 TI - Genetically modified NT2N human neuronal cells mediate long-term gene expression as CNS grafts in vivo and improve functional cognitive outcome following experimental traumatic brain injury. AB - Human Ntera-2 (NT2) cells can be differentiated in vitro into well-characterized populations of NT2N neurons that engraft and mature when transplanted into the adult CNS of rodents and humans. They have shown promise as treatments for neurologic disease, trauma, and ischemic stroke. Although these features suggest that NT2N neurons would be an excellent platform for ex vivo gene therapy in the CNS, stable gene expression has been surprisingly difficult to achieve in these cells. In this report we demonstrate stable, efficient, and nontoxic gene transfer into undifferentiated NT2 cells using a pseudotyped lentiviral vector encoding the human elongation factor 1-alpha promoter and the reporter gene eGFP. Expression of eGFP was maintained when the NT2 cells were differentiated into NT2N neurons after treatment with retinoic acid. When transplanted into the striatum of adult nude mice, transduced NT2N neurons survived, engrafted, and continued to express the reporter gene for long-term time points in vivo. Furthermore, transplantation of NT2N neurons genetically modified to express nerve growth factor significantly attenuated cognitive dysfunction following traumatic brain injury in mice. These results demonstrate that defined populations of genetically modified human NT2N neurons are a practical and effective platform for stable ex vivo gene delivery into the CNS. PMID- 12722831 TI - Colocalization of tau and alpha-synuclein epitopes in Lewy bodies. AB - The major protein constituent of Lewy bodies (LBs), the pathological hallmark of Parkinson disease and dementia with Lewy bodies, is considered to be alpha synuclein, but other proteins, in particular the microtubule-associated protein tau, have been implicated in the pathogenesis of LBs. Tau is the major structural component of neurofibrillary tangles (NFTs). Both direct immunochemical studies of partially purified LBs and indirect immunohistochemical studies have suggested that LBs may contain tau, but most of these studies were based upon a single tau antibody, and immunologic cross-reactivity was not completely excluded. To gain insight into the relation between tau and alpha-synuclein in LBs, double immunostaining was performed in Lewy body cases with a rabbit polyclonal antibody to alpha-synuclein and a panel of monoclonal antibodies to phospho- and nonphospho-tau epitopes (Alz50, CP9, CP13, PG5, TG3, PHFI) that spanned the length of the tau molecule. Tau-immunoreactive LBs were present in the medulla in 80% of the cases, irrespective of Braak stage. All tau antibodies recognized at least some LBs, arguing against nonspecific antibody cross-reactivity. In most lesions the tau immunostaining was present at the periphery of the LB. The phospho-tau antibody, TG3, detected more LBs than any of the other tau antibodies. The proportion of LBs with tau immunoreactivity was greatest in neurons vulnerable to NETs, such as those in the locus ceruleus and basal nucleus of Meynert, and least in neurons resistant to NFTs, such as the dorsal motor nucleus of the vagus in the medulla. The present results suggest that tau may coaggregate with alpha-synuclein in LBs, especially in neuronal populations vulnerable to both NFTs and LBs. PMID- 12722833 TI - Regulation and functional effects of monocyte migration across human brain derived endothelial cells. AB - We have used human brain-derived endothelial cells (HBECs) maintained under basal culture conditions in a Boyden chamber assay system as an in vitro model of migration of cells of systemic immune origin across the blood brain barrier (BBB) during the initiation of a CNS-directed inflammatory response. In this study we evaluated the molecular mechanisms that regulate passage of ex vivo peripheral blood-derived monocytes across this barrier and the effects of such migration on the properties of both the HBECs and the monocytes. Our results indicate that monocytes can migrate across HBECs in the absence of inflammatory conditions, at rates exceeding those of lymphocytes. Monocyte migration could be significantly inhibited by the addition of blocking antibodies to intercellular adhesion molecule (ICAM)-1, very late antigen (VLA)-4 integrin, and monocyte chemoattractant protein (CCL-2/MCP-1), or treatment with tissue inhibitor of metalloproteinase (TIMP-1). Following monocyte migration there was a significant increase in permeability of soluble molecules and an enhanced rate of T cell migration across HBECs. The enhanced permeability could be partially prevented with anti-TNF-alpha antibody. The migration process did not induce the upregulation of either co-stimulatory molecules or chemokine receptors on the monocytes. These studies emphasize the functional role of monocyte-endothelial interactions in permitting target access of a CNS-directed cell-mediated immune response. PMID- 12722835 TI - The discovery of DNA structure: 50 years on. PMID- 12722834 TI - Downregulation of Bcl-2 proteins in type I spinal muscular atrophy motor neurons during fetal development. AB - Spinal muscular atrophy (SMA) is an autosomal recessive disorder caused by mutations in the survival motor neuron gene. The degeneration and loss of the anterior horn cells constitute the major neuropathological finding in SMA, although the mechanism and timing of this abnormal motor neuron death remain unknown. It has recently been reported that the fetal SMA spinal cord shows a significant increase in cells with DNA fragmentation, suggesting that the programmed cell death is aberrantly increased in type I SMA during development. We have analyzed 2 antiapoptotic proteins, Bcl-2 and Bcl-X, by Western blot and immunohistochemistry screening for differential expression in control and SMA fetal spinal cords. Expression of these proteins was found in various neuronal populations and structures of the developing spinal cord. At 15 weeks, motor neurons of SMA fetuses showed a marked decrease in the levels of Bcl-2 and a delay in the expression of Bcl-X in comparison with controls. The difference in the pattern and degree of expression is consistent with a role for both proteins in the aberrant programmed cell death observed in type I SMA. PMID- 12722832 TI - Schwann cell apoptosis in the postnatal axotomized sciatic nerve is mediated via NGF through the low-affinity neurotrophin receptor. AB - Schwann cell death is a developmentally regulated phenomenon and is also induced after peripheral nerve axotomy in neonatal rodents. In this study, we explored whether ligand-induced activation of the low-affinity neurotrophin receptor (p75(NTR)) is responsible for inducing Schwann cell death in vivo. Administration of exogenous nerve growth factor (NGF) to the axotomized nerve site in wild-type animals resulted in a 2.6-fold increase in Schwann cell apoptosis in the distal nerve stumps compared to axotomy alone. No increase in apoptosis, above baseline levels, was seen in p75(NTR)-mutant mice either with or without NGF When anti-NGF antibodies were administered to the site of the peripheral nerve lesion in wild type mice there was a reduction in the percentage of Schwann cell apoptosis to levels seen in both the quiescent state and in the axotomized nerves of the p75(NTR)-mutant mice. These results demonstrate that apoptosis of Schwann cells in axotomized peripheral nerve is mediated predominantly through p75(NTR) signaling and initiated via endogenously produced NGF. PMID- 12722836 TI - Institute of Human Development, Child and Youth Health. PMID- 12722838 TI - Research subject advocates: to whom are they loyal? PMID- 12722837 TI - Proinflammatory cytokine levels in hyperthyroidism. AB - BACKGROUND: Among suspected causes of the osteoporosis frequently seen in untreated thyrotoxicosis are the osteotrophic cytokines. We studied serum levels of osteotrophic cytokines including interlukin (IL)-1, IL-6, IL-8 and tumour necrosis factor alpha (TNF-alpha) in patients with various hyperthyroid states. METHODS: Serum cytokines were detected in 4 groups of SUBJECTS: 14 patients with Graves' disease, 9 patients with toxic nodular goitre, 27 patients with toxic multinodular goitre and 30 euthyroid control subjects. The levels of IL-1-beta, IL-6, IL-8 and TNF-alpha in the serum were determined by the IMMULITE autoanalyzer, using a chemoilluminesence method. RESULTS: Compared with euthyroid control subjects, patients with hyperthyroidism had significantly elevated serum levels of IL-8 (506.8 pg/mL v. 7.0 pg/mL, p < 0.001) and TNR-alpha (18.6 pg/mL v. 8.7 pg/mL, p < 0.05). Levels of IL-1-beta (12.2 pg/mL v. 5.0 pg/mL) and IL-6 (30.3 pg/mL v. 5.3 pg/mL) were also higher in controls, but the differences were not statistically significant. Levels of the cytokines were similar in 14 patients with diffuse goitre compared with 36 patients having nodular goitre. Cytokine levels in 20 premenopausal and 20 postmenopausal women with hyperthyroidism were also similar. INTERPRETATION: We conclude that increased circulating cytokine concentrations observed in patients with hyperthyroidism may derive from the activation of humoral reactions in sites other than the thyroid. PMID- 12722839 TI - Bioinformatics: implications for medical research and clinical practice. PMID- 12722841 TI - The population approach to stroke prevention: a Canadian perspective. AB - OBJECTIVES: To contrast the population and clinical approaches to stroke prevention and to review Canadian data relevant to estimating disease burden, assessing risk factors, designing preventive strategies and organizing health services. METHOD: A narrative review of the published literature and statistical data accessible through the Internet. MAIN FINDINGS: Unlike the clinical approach, which emphasizes individual patients at high risk diagnosed and treated intensively, usually by medical or surgical means (or both), a population approach focuses on the entire population and bases interventions on behavioural and environmental changes. Stroke offers a particularly promising target for prevention. It represents a leading cause of serious disability, death and reduced quality of life. The aging of our population threatens to increase the already considerable burden. Stroke shares several risk factors with other chronic diseases, especially ischemic heart disease. These risk factors vary in their impact on Canadians (population attributable risk proportion), which is a function of their prevalence and strength of association with the occurrence of stroke. Although effective preventive measures are available for people at high risk, they are not being applied systematically among potential beneficiaries. Small reductions in the exposure to risk factors in the entire population offer an alternative, where even modest success may translate into major gain. CONCLUSIONS: The clinical and population approaches to stroke prevention are complementary. Existing national strategies directed at promotion of healthy life styles (especially physical activity) and hypertension control, when fully implemented, will reduce the frequency, severity and impact of stroke on Canadian society. PMID- 12722840 TI - Bioinformatics in research and clinical practice. PMID- 12722842 TI - Family physicians are an important source of mental health care. AB - While comprising about 15 percent of the physician workforce, family physicians provided approximately 20 percent of physician office-based mental health visits in the United States between 1980 and 1999. This proportion has remained stable over the past two decades despite a decline in many other types of office visits to family physicians. Family physicians remain an important source of mental health care for Americans. PMID- 12722843 TI - Differentiating foot fractures from ankle sprains. PMID- 12722844 TI - Management of risk factors in relatives of patients with SAH. PMID- 12722846 TI - Screening and intervening for patients with substance use disorders. PMID- 12722845 TI - Evaluation and treatment of heat-related illnesses. PMID- 12722847 TI - Hormone therapy: continuing discussion and debate. PMID- 12722848 TI - Prevention and treatment of common eye injuries in sports. AB - Sports have become increasingly popular and account for numerous eye injuries each year. The sports that most commonly cause eye injuries, in order of decreasing frequency, are basketball, water sports, baseball, and racquet sports. Sports are classified as low risk, high risk, and very high risk. Sports-related eye injuries are blunt, penetrating, and radiation injuries. The use of eye protection has helped to reduce the number and severity of eye injuries. The American Society for Testing and Materials has established performance standards for selected eyewear. Consultation with an eye care professional is recommended for fitting protective eyewear. The functionally one-eyed, or monocular, athlete should take extra precautions. A preparticipation eye examination is helpful in identifying persons who may be at increased risk for eye injury. Sports-related eye injuries should be evaluated on site with an adequate examination of the eye and adnexa. Minor eye injuries may be treated on site. The team physician must know which injuries require immediate referral to an ophthalmologist and the guidelines for returning an athlete to competition. PMID- 12722849 TI - Information from your family doctor. Eye injuries in sports. PMID- 12722850 TI - Nocturnal enuresis. AB - Nocturnal enuresis is a common problem that can be troubling for children and their families. Recent studies indicate that nocturnal enuresis is best regarded as a group of conditions with different etiologies. A genetic component is likely in many affected children. Research also indicates the possibility of two subtypes of patients with nocturnal enuresis: those with a functional bladder disorder and those with a maturational delay in nocturnal arginine vasopressin secretion. The evaluation of nocturnal enuresis requires a thorough history, a complete physical examination, and urinalysis. Treatment options include nonpharmacologic and pharmacologic measures. Continence training should be incorporated into the treatment regimen. Use of a bed-wetting alarm has the highest cure rate and the lowest relapse rate; however, some families may have difficulty with this treatment approach. Desmopressin and imipramine are the primary medications used to treat nocturnal enuresis, but both are associated with relatively high relapse rates. PMID- 12722851 TI - Information from your family doctor. Bed-wetting. PMID- 12722852 TI - Osteoporosis in men. AB - Osteoporosis in men is now recognized as an increasingly important public health issue. About 30 percent of hip fractures occur in men, and one in eight men older than 50 years will have an osteoporotic fracture. Because of their greater peak bone mass, men usually present with hip, vertebral body, or distal wrist fractures 10 years later than women. Hip fractures in men, however, result in a 31 percent mortality rate at one year after fracture versus a rate of 17 percent in women. Major risk factors for osteoporosis in men are glucocorticoid use for longer than six months, osteopenia seen on plain radiographs, a history of nontraumatic fracture, hypogonadism, and advancing age. Bisphosphonates and teriparatide (recombinant parathyhroid hormone) have recently been approved for use in men and should be considered along with supplemental calcium and vitamin D. Increased awareness by physicians of risk factors for male osteoporosis--and early diagnosis and treatment--are needed to decrease the morbidity and mortality resulting from osteoporotic fractures. PMID- 12722853 TI - Recognition of alcohol and substance abuse. AB - Ten percent of the population abuses drugs or alcohol, and 20 percent of patients seen by family physicians have substance-abuse problems, excluding tobacco use. These patients can be identified by relying on regular screening or a high index of suspicion based on "red flags" that can be noted in various clinical situations. The modified CAGE questionnaire is an excellent screening instrument, but several alternatives are available. The best screening test is one that the physician will routinely use well. Laboratory indicators such as gamma-glutamyl transpeptidase, mean corpuscular volume, and carbohydrate-deficient transferrin are nonspecific but can add to the evidence of alcohol abuse. If problem alcohol use is diagnosed, even brief physician advice can be helpful. If the problem has progressed to addiction, referral to an addiction specialist or treatment center is recommended. Special issues arise in dealing with substance abuse in adolescents, elderly patients, and patients with mental illness, but the family physician can play an important role in recognizing this common problem. PMID- 12722854 TI - Information from your family doctor. Substance abuse--how to recognize it. PMID- 12722855 TI - Fusiform excision. AB - The fusiform excision technique is commonly used by physicians for removing skin and subcutaneous lesions. The technique requires basic skills in anesthetic administration, lesion excision, and suture closure. Physicians experienced in skin surgery can often perform the procedure unsupervised after two to five precepted sessions. The fusiform excision technique should be used to biopsy any suspicious pigmented lesions because prognosis may depend on the depth of the lesion. A properly designed fusiform excision has a length-to-width ratio of 3 to 1 and produces a 30-degree angulation at both edges of the wound. The skin edges should be everted during the closure to improve final scar appearance. Malignant growths may require a second procedure to provide a wider margin of excision around the lesion. Post-excisional wound infections are uncommon with use of proper aseptic technique. PMID- 12722856 TI - Information from your family doctor. Fusiform excision procedure. PMID- 12722858 TI - Screening for depression. PMID- 12722859 TI - Photo quiz. A nonhealing ulcer of the hand. PMID- 12722860 TI - Information from your family doctor. Abstinence: information for teens. PMID- 12722861 TI - Information from your family doctor. Birth control. PMID- 12722862 TI - Information from your family doctor. Progestin-only contraceptives. PMID- 12722868 TI - Should I give money to my patients? PMID- 12722867 TI - ACOG releases guidelines on diagnosis and management of polycystic ovary syndrome. PMID- 12722870 TI - Celsion microwave therapy proves effective in breast cancer. PMID- 12722869 TI - Taking stock of targeted therapy in cancer: rational expectations or irrational exuberance? PMID- 12722871 TI - Encouraging results for Plenaxis in prostate cancer trial. PMID- 12722872 TI - Aplidin increases sensitivity to treatment in leukemia and lymphoma cells. PMID- 12722873 TI - Role of capecitabine (Xeloda) in breast cancer. AB - One rationale for the development of new treatment strategies for advanced breast cancer is to provide targeted antineoplastic therapy, while at the same time improving the quality of life of patients. One such drug, capecitabine (Xeloda), is an oral fluoropyrimidine 5-fluorouracil carbamate. Capecitabine is converted to 5-fluorouracil primarily in cancer tissue and it has been demonstrated to combine ease of administration, a manageable toxicity profile and potent antineoplastic activity. Capecitabine is widely used in metastatic breast cancer and offers symptom palliation and in combination with docetaxel (Taxotere) improved survival compared with docetaxel alone. Its toxicity profile includes hand-foot syndrome and stomatitis and diarrhea, whereas its hematologic side effects are mild. Capecitabine has been evaluated as a single agent in women with advanced breast cancer where it offers an overall response rate of 20-30%. Capecitabine is synergistic with other chemotherapeutic agents, such as the taxanes, where it increases the response rate to over 40%. This review will place the available data on the use of capecitabine in metastatic breast cancer as a single agent or as part of a combination regime in context. PMID- 12722874 TI - Pemetrexed (Alimta): a novel multitargeted antifolate agent. AB - Pemetrexed (Alimta) is a novel, multitargeted antifolate that inhibits at least three of the enzymes involved in folate metabolism, and purine and pyrimidine synthesis. These enzymes are thymidylate synthase, dihydrofolate reductase and glycinamide ribonucleotide formyltransferase. Pemetrexed has demonstrated broad antitumor activity in Phase II trials in a wide variety of solid tumors, including mesothelioma, non-small cell lung, breast, cervical, colorectal, head and neck, and bladder cancers. Promising activity has also been demonstrated when pemetrexed is combined with cisplatin and gemcitabine (Gemzar). A pivotal Phase III study in mesothelioma has been presented. This study indicates the superiority of pemetrexed in combination with cisplatin versus cisplatin alone in this disease. The most significant toxicities of pemetrexed, myelosuppression and mucositis have been significantly ameliorated by folate and vitamin B12 supplementation. More importantly, vitamin supplementation has not demonstrated any adverse efficacy. This review discusses the biochemistry and clinical activity of pemetrexed. PMID- 12722876 TI - Improved tolerability of chemotherapy in soft tissue sarcomas: old and new strategies. AB - Anthracyclines and alkylating agents (ifosfamide, Mitoxana) are the mainstays of chemotherapy for soft tissue sarcoma and as there is a close correlation between dose-intensity and response, methods should be developed to ameliorate the clinical tolerability of these agents. Improvements in the efficacy of medical treatments for soft tissue sarcomas may derive from old strategies by aiming to counteract the side effects of standard chemotherapy regimes and from new, less toxic, anticancer drugs. This paper reviews the currently available options for reducing the cardiotoxicity of anthracyclines, the role of growth factors and autologous stem cell transplantation in dose-intensification of chemotherapy and also examines the clinical impact of the more promising new agents. PMID- 12722875 TI - Zoledronic acid (Zometa) use in bone disease. AB - Zoledronic acid (Zometa) is the most recent addition to the clinically available bisphosphonates. Clinical benefits in metabolic, as well as cancer-related bone disease have been observed. In addition to its profound antiosteoclast effects, it has demonstrated anticancer effects in preclinical models. Zoledronic acid has been evaluated in randomized, double-blind clinical trials of osteoporosis, Paget's disease of bone, and metastatic, osteolytic and osteoblastic bone disease. Antiosteoclast activity has been demonstrated by reductions in the bone breakdown products N-telopeptide, C-telopeptide and deoxypyridinoline. Bone mineral density, measured by dual energy x-ray absorptometry, is increased with administration of zoledronic acid in postmenopausal osteoporosis. Clinical benefit in cancer includes improvement in bone pain, reductions in skeletal events and delay in time-to-first-skeletal-events. These zoledronic acid treatment benefits have been demonstrated in patients with multiple myeloma, breast, prostate and lung cancer, and other solid tumors. PMID- 12722877 TI - Recent advances in systemic therapy of soft tissue sarcomas. AB - Improvements in surgical techniques, for example reconstructive surgery; and radiation therapy techniques, such as intensity-modulated radiotherapy; have made some impact on the functional outcomes of sarcomas with a local biology. Effective local control can be achieved with effective function in the vast majority of patients. The problem of distant metastases, however, continues to plague a large group of patients with this disease. Recent advances in the systemic therapy of sarcomas is highlighted by the rapid development and approval of the molecularly targeted therapy imatinib (Gleevec) for advanced gastrointestinal stromal tumors. Several other agents have been, or are being studied with far less rewarding results. Among these, the more encouraging examples include the nucleoside analog gemcitabine (Gemzar) and gemticibine/docetaxel (Taxotere) in combination, which display some selective activity in sarcomas of gynecologic origin. The marine compound ecteinascidin (ET 743) has been studied in two different schedules (24 and 3 h infusions), demonstrating biological activity worthy of further investigation. Identification of new agents with activity in this diverse group of diseases is extremely important. Identification of specific targets responsible for tumorigenesis and effective inhibition of these targets holds the most promise for future improvement in cure rates. However, until such time, it is equally important to emphasize clinical research attempting to further optimize the use of the standard chemotherapeutic agents, with growth-factor support in appropriately selected patients. PMID- 12722878 TI - Advances in the adjuvant treatment of infantile fibrosarcoma. AB - Infantile fibrosarcoma is a rare soft tissue tumor, predominately affecting young infants. It grows rapidly and is locally infiltrative but rarely metastases. Complete surgical removal is usually curative but is impossible in some patients and would result in significant functional or cosmetic consequences in many others. Neoadjuvant chemotherapy will cause many tumors to shrink significantly, allowing less mutilating surgical resections to be performed--this is the current recommendation where immediate surgical removal cannot be accomplished without unacceptable morbidity. In contrast, there is no defined role for adjuvant chemotherapy or radiation following complete surgical resection. Although there is a significant risk of local recurrence, most of these can be successfully treated with further surgery and the overall survival rate exceeds 90%. PMID- 12722879 TI - Aromatase inhibitors for treatment of postmenopausal patients with breast cancer. AB - The third generation of aromatase inhibitors and inactivators, such as anastrozole (Arimidex), letrozole (Femara) and exemestane (Aromasin), have become available for treatment of postmenopausal breast cancer patients. Several clinical trials have demonstrated that these new drugs can achieve better treatment results than megestrol acetate (Megace) and may replace tamoxifen for the first-line hormonal therapy for metastatic breast cancer patients. In fact, these drugs are now used in many hospitals and clinics for patients with metastatic breast cancer who were previously given tamoxifen as adjuvant treatment. However, the primary concern is whether they can be used as first-line agents for adjuvant treatment of primary breast cancer or are suitable for breast cancer prevention in view of possible adverse side effects. Recently, the Arimidex and Tamoxifen Alone or in Combination trial demonstrated the superiority in terms of disease-free survival of anastrozole over tamoxifen in adjuvant use for postmenopausal patients with Stage I and II primary breast cancer. The results of this report indicate the potential of anastrozole as an alternative drug in the adjuvant setting, although the mean follow-up time is so far only 47 months. Additional data regarding survival resulting from comparative trials of letrozole and tamoxifen and of exemestane and tamoxifen are expected to be available in a few years. However, limited information is available regarding adverse events caused by long-term administration of aromatase inhibitors. Longer follow-up is needed to determine the efficacy and safety of these new aromatase inhibitors when used for adjuvant treatment of postmenopausal patients with breast cancer. PMID- 12722880 TI - Chemoprevention of prostate cancer. AB - Dietary factors and other naturally occurring substances may emerge as potent therapeutic or preventative agents in the battle against prostate cancer. Much of the current support for these agents is epidemiologically based, but new prospective studies are now underway which may support their use in conventional medical practice. PMID- 12722881 TI - von Hippel-Lindau disease: recent advances and therapeutic perspectives. AB - von Hippel-Lindau disease is a hereditary cancer syndrome predisposing carriers to the development of a panel of highly vascularized tumors such as central nervous system and retinal hemangioblastomas, endolymphatic sac tumors, clear cell renal cell carcinomas, pheochromocytomas and pancreatic neuroendocrine tumors. The disease is the foremost cause of inherited renal cell carcinomas, which is induced by germline mutations of the VHL tumor-suppressor gene also inactivated in most sporadic renal cell carcinomas. VHL appears to be a pivotal gene in the oxygen-sensing pathway, mainly involved in targeting the hypoxia inducible factors for ubiquitination. This discovery is opening the way for the development of new specific drugs inhibiting hypoxia-inducible factors and/or their downstream targets, possibly representing an attractive treatment not only for von Hippel-Lindau disease but also for sporadic renal cell carcinomas and others cancers. PMID- 12722882 TI - Role of systemic chemotherapy in metastatic cervical cancer. AB - Cervical cancer is a chemoresponsive tumor. Concurrent chemotherapy with cisplatin and radiotherapy has resulted in improved survival in patients with locally advanced cervical cancer and is now standard of care. There are many active drugs in metastatic cervical cancer with cisplatin being the most active single agent. Although response rates are significantly higher with cisplatin combinations, to date, there is no evidence to suggest this is associated with an improved survival. However, this is still an area that is attracting much research interest. The role of chemotherapy in metastatic cervical cancer will be reviewed together with some of the new areas of research. PMID- 12722883 TI - Treatment of tumor-induced hypercalcemia: a solved problem? AB - Less than 25 years ago, tumor-induced hypercalcemia was often a lethal complication of cancer. Nowadays, it can be successfully and easily treated in at least 90% of the cases by rehydration and potent antiosteoclastic bisphosphonates. The standard therapy consists of the administration of 90 mg of pamidronate (Aredia Dry Powder) or more recently, 4 mg of zoledronic acid (Zometa)], which is even more efficient, at least in patients without bone metastases. Recurrent hypercalcemia is nevertheless difficult to control and antibodies against parathyroid-hormone-related protein may prove to be a useful treatment. PMID- 12722885 TI - The role of psychoanalytic theory and practice in understanding and treating schizophrenia: a rejoinder to the PORT report's condemnation of psychoanalysis. AB - The Schizophrenia PORT report rejects, in recommendation 22, the use of psychodynamic therapies. It also rejects, in recommendation 26, a crucial point of psychoanalytic theory by rejecting family interventions based on the assumption that the family of origin can make a ca usal contribution to schizophrenia. Both recommendations are based on level "C" evidence, which is defined by the authors as a: "Recommendation based primarily on expert opinion, with minimal research-based evidence, but significant clinical experience" (Lehman et al., 1998, p. 2). Conclusions based on low-level evidence would be better formulated in tentative rather than categorical statements as is now done in the PORT report. More substantially, I report on a number of empirical studies that contradict the claims of recommendations 22 and 26. I therefore urge that the two PORT recommendations be revised. I argue that the recommendations can be rewritten so that a useful point in each of the recommendations can be preserved and even strengthened. With reference to recommendation 22, it is wise to stress that psychoanalytic-inspired therapies with schizophrenics should not simply apply the same methods used in the treatment of neurotics. With reference to recommendation 26, it is important to stress that imposing or augmenting guilt is not a good therapeutic technique. Similarly, it is not good therapeutic practice to deny the truth. To tell the truth and not create guilt requires artful interventions. If recommendations 22 and 26 were to be rewritten as suggested, they would become scientifically defensible and would make a positive contribution by stressing the contributions that many approaches, including psychoanalysis, can make to the treatment of schizophrenia. As they stand now, recommendations 22 and 26 are contradicted by the scientific evidence I will submit. PMID- 12722884 TI - FRONTLINE--the schizophrenic person and the benefits of the psychotherapies- seeking a PORT in the storm. PMID- 12722886 TI - Violent behavior in chronic schizophrenia and inpatient psychiatry. AB - The author describes how work with inpatients with chronic schizophrenia has contributed to a better understanding of antisocial behavior. She has used the concept of regression, along biological and psychological lines, to hypothesize fantasies of primitive object relationships which drive the behavior. Engaging patients in thoughtful reflection, she has introduced a third perspective on the potential state of mind of the important people in their lives; the possibility of a concerned object, rather than that of a vengeful or rejecting object. Finding that even those with resistant schizophrenia respond with change in behavior, she found that she could more easily employ the same psychoanalytic concepts in engaging those who present with more acute problems of violent and suicidal behavior. PMID- 12722887 TI - The mask of psychotic diagnoses. AB - Chronic mental illness results in the patient becoming adhered to a DSM-IV diagnostic label. Over time, this diagnosis can expand and become a "mask" that invisibly covers over the true person of the patient. Most commonly, two things then occur. First, the outside world forgets that the patient is a person and family, friends, staff, and doctors begin to treat the patient according to the superficial aspects of what the mask of the diagnosis connotes, rather than connecting with the person struggling with the illness. Second and, perhaps, more insidious, is that the patient, who has been vulnerable and shattered by his or her experience and battle with the illness, adopts the mask as a kind of invisible protective shield. The task of making contact with the patient behind the mask of the diagnosis is therefore a formidable one for psychoanalysts and therapists and staff who work with seriously ill patients. Treatment must focus on the dual process of interfering with the patient's use of the diagnostic mask while, at the same time, making safe contact with the person of the patient behind the mask. A focus on affect can help achieve these dual goals. By utilizing Semrad's (Semrad and van Buskirk, 1969) method of noticing and asking about "feelings" as conveyed by hallucinations, delusions, or bodily sensations, a reliable relationship can evolve and the clinician can come to have an important "selfobject" (Kohut, 1971) meaning for the patient. By attuning to the patient's "vitality" affects (Stern, 1985), great stability and a new sense of "aliveness" is made possible to help the patient emerge from the deadening effects of the illness and the mask of the diagnosis. PMID- 12722888 TI - Pre-therapy: a newer development in the psychotherapy of schizophrenia. AB - Pre-Therapy is an evolution from client-centered theory and practice. The focus of Pre-Therapy is on regressed patients such as the psychotic retarded or chronic schizophrenics. It is also applicable to demented populations. The central construct is psychological contact, described as the necessary condition of psychotherapy and outlined on three levels: (1) the contact reflections--the "work" the therapist does; (2) the contact functions--the psychological process of the patient; (3) the contact behaviors--operationalized behavior for measurement. The Pre-Expressive Self, an intuitive and heuristic concept, interprets the progress from pre-expressive to expressive states of communication that is necessary for psychotherapy. PMID- 12722889 TI - Interview with Gaetano Benedetti, M.D. AB - Professor Gaetano Benedetti has been working as a psychoanalyst with individuals with a schizophrenic disorder for over 50 years in his capacity as psychotherapist, supervisor, and teacher. In this interview Professor Benedetti defines the basic psychological problems in schizophrenia. He and his younger colleague in the field of schizophrenia, Dr. Maurizio Peciccia, understand the illness to be the result of an interaction between possible neurobiological vulnerabilities, overwhelming affects, and a self lacking cohesion and integration. Specifically, they characterize the core psychological deficit in schizophrenia to be a deintegration of the separate and symbiotic selves of the patient, resulting in the oscillation between pathological symbiosis with the world and a defensive autistic-like retreat from object relations. Other topics discussed include: the therapeutic symbiosis; the transitional subject; progressive psychopathology; therapeutic counteridentification; the origins of schizophrenia; therapeutic images and psychotherapeutic transformation. PMID- 12722890 TI - The tragedy of schizophrenia without psychotherapy. AB - No one who read Frieda Fromm-Reichmann's "Transference Problems in Schizophrenia" could reasonably think about persons with schizophrenia in the same way as before. Her writings made clear that schizophrenia is a human experience with meaning, meaning that is hard to uncover, but it only takes patience, kindness, a tolerance for not understanding as well as for the patient's desperate defenses, and a willingness to understand the human condition at its most painful and to take psychoanalytic ideas seriously when patients talk about them. Understanding persons with schizophrenia means facing facts about ourselves, our families, and our society that we do not want to know, or to know again (in the case of repressed feelings and experiences). Families and professionals are settling for treatments that aim at making the patient a lifelong cripple who is not too disturbing. Psychoeducational programs, which potentially could be helpful, usually give false information which makes worse the burdens of both patients and their families. The ultimate genetic experiment, the Nazi sterilization and annihilation of patients, led to no decrease in schizophrenia in the next generation. Long-term follow-up studies show one third of schizophrenics fully recover within 25 years and another third have social recoveries with or without treatment--and psychological treatments before the neuroleptic era, from "moral treatment" to psychoanalytic therapies, produced superior results, but we are not using them. The central role of terror in producing symptoms and the genesis and psychotherapeutic handling of symptoms, including delusions and hallucinations, will be described. PMID- 12722891 TI - Lessons in how to ruin a study in psychotherapy effectiveness: a critical review of the follow-up study from Chestnut Lodge. AB - The principal publications from the follow-up study of former patients at Chestnut Lodge are reviewed and critiqued. Originally, the follow-up study was intended as a study of the psychotherapeutic process and its outcome. While widely referred to as a follow-up study, it is not possible to make conclusions about psychotherapy from the study. Data about the psychodynamic aspects of therapy have been collected but never analyzed. Additionally, the study does not fulfill the requirements of a psychotherapeutic outcome study. The conclusion of the current review is that as the study turned out, it only allows one to draw conclusions about different kinds of illness courses and predictor factors. PMID- 12722892 TI - Evidence-based psychosocial treatment practices in schizophrenia: lessons from the patient outcomes research team (PORT) project. AB - The care of persons with schizophrenia, the prototypical severe mental illness, has been a barometer of mental health care policy for decades. The prevalence, severity, and costs of schizophrenia combine to make this illness a major health problem throughout the world. In 1992, the Agency for Health Care Policy and Research and the National Institute of Mental Health funded the Schizophrenia Patient Outcomes Research Team (PORT) at the University of Maryland School of Medicine and the Johns Hopkins University School of Public Health. The PORT undertook several activities, including a comprehensive review of the empirical literature on the treatment of persons with schizophrenia; development of evidence-based treatment recommendations; description of current treatment practices; and comparison of these current practices to the evidence-based treatment recommendations, using administrative claims data and a survey of persons under treatment for schizophrenia; and dissemination of the treatment recommendations to evaluate impacts on practices. The PORT found that despite considerable evidence for effective treatments for persons with schizophrenia, most patients do not receive an appropriately comprehensive treatment "package." In particular, efficacious psychosocial treatments are highly underutilized. PMID- 12722893 TI - Psychological treatments for psychosis: history and overview. AB - This article is part of the ISPS (International Society for the Psychological Treatment of the Schizophrenias and other Psychoses) task force report on the PORT (Patients Outcome Research Team) recommendations for treatment of schizophrenia. It reviews psychological treatment approaches in psychosis to date and assesses recent trends. The most influential therapies have been psychoanalytic/psychodynamic, cognitive behavioral (CBT), and supportive therapy. PMID- 12722894 TI - Evidence-based medicine in the psychological treatment of schizophrenia. AB - Treatments should be based on reviews of the best available evidence. However, there are different views about the types of evidence that may be included. Evidence-based medicine (EBM) has its own clear hierarchy with systematic reviews of randomized controlled trials (RCTs) at the top level and expert and user opinion at the lowest level of evidence. Other paradigms, such as qualitative approaches, are difficult to place in such a hierarchy. This article examines some of the assumptions behind evidence-based practice, particularly when applied to complex "real life" clinical situations. A case example is used to demonstrate some of the complexity of applying evidence to practice. A complementary paradigm of practice-based evidence is described. PMID- 12722896 TI - The concept of schizophrenia and phase-specific treatment: cognitive-behavioral treatment in pre-psychosis and in nonresponders. AB - The development of psychosis is often described as a process that goes through different linear stages such as a premorbid and a prodromal phase, with the onset of psychosis as the end state. In this article we elaborate a hierarchical model in which psychosis is understood at different levels of abstraction: we differentiate an etiological, a descriptive, and a therapeutical level, and finally a level of understanding. We regard coping as a special psychological process that starts when symptoms of psychosis first emerge (often many years before treatment is initiated) and continues throughout therapy. In patients who do not respond very well to standard treatment (approximately 15-20% of all patients who suffer from schizophrenia), coping strategies are probably some of the most important long-term prognostic factors. Two cognitive-oriented treatment programs are described: the German model for possible pre-psychosis and the UK tradition for treatment of chronic psychotic symptoms (nonresponders). We conclude that psychological treatment that focuses on coping strategies might be of help both in possible pre-psychotic stages and in nonresponders. PMID- 12722895 TI - A critique of the methods and conclusions in the patient outcome research team (PORT) report on psychological treatments for schizophrenia. AB - The schizophrenia Patient Outcome Research Team (PORT; Lehman and Steinwachs, 1998), a policy paper on the treatment of schizophrenia sponsored by the United States government Agency for Health Care Policy and Research, has minimized the efficacy and utility of psychological treatments for schizophrenia and concluded that some treatments like psychoanalytic psychotherapy are contraindicated and harmful. A critical review of Recommendations 22, 23, 24, 25, and 26 (those addressing the efficacy of psychological treatments) was undertaken. The purpose of this critique was to describe the threats to validity inherent in the PORT report's design and to demonstrate how those threats make the relevant recommendations specious. In addition, empirical evidence from meta-analytic reviews is described to show that, indeed, psychological treatments for schizophrenia are efficacious. Limitations of the available evidence and suggestions for a PORT revision are discussed. PMID- 12722897 TI - Psychological treatment in pre- and early psychosis. AB - This article aims to overview a broad range of psychosocial treatments for first episode psychosis, and for the prodromal phase (or so-called at-risk mental state)--the period preceding the first acute episode (Yung and McGorry, 1996). Firstly, an introduction to the empirically based rationale for early intervention in first-episode psychosis is provided. This is followed by a selective review of individual psychotherapies for early psychosis, which then proceeds to a discussion of family-based interventions for first-episode families and the role of group programs. Next, the role of psychological interventions within the newly emerging indicated preventive approach (Mrazek and Haggerty, 1994) for at-risk mental state is examined before some illustrative case material is presented. It is concluded that integrated psychosocial interventions for first-episode psychosis and for prodrome are newly emerging, innovative fields that offer some preventive opportunities. These opportunities, combined with some initial outcome data, warrant continuing research and clinical innovation. PMID- 12722898 TI - Psychological trauma and psychosis: another reason why people diagnosed schizophrenic must be offered psychological therapies. AB - This article summarizes the research literature documenting the high prevalence of psychological trauma, including childhood sexual and physical abuse, among people diagnosed psychotic in general and schizophrenic in particular. A review of the relevant literature indicates that childhood trauma may in some cases be causally related to the development of psychotic symptomatology later in life, perhaps by contributing to the diathesis in the stress-diathesis equation. However it is argued that regardless of whether clinicians view the relationship as causal, contributory, co-morbid, or coincidental is irrelevant to the fact that this high prevalence dictates that all people diagnosed schizophrenic receive a proper trauma assessment and, where appropriate, be offered psychological treatments to address the sequelae of the trauma or abuse. Taken in conjunction with the proven effectiveness of various psychological and psychosocial treatments in ameliorating the symptomatology and improving the quality of life of people diagnosed schizophrenic, the need to address the trauma found in such high rates in this population requires a broadening, and not as some suggest a limitation, of the range of treatments offered. The authors identify and challenge ideologically, rather than empirically, driven assumptions about the etiology of schizophrenia, particularly the distorted but dominant version of the diathesis-stress model, which claims that the diathesis is predominantly or exclusively biogenetic in origin and therefore inaccurately places all psychosocial stressors and traumas exclusively on the stress side of the equation. It is this bias that inhibits many clinicians from taking trauma histories with people diagnosed schizophrenic and from offering appropriate treatment when trauma is disclosed. It is the same bias that leads some to argue for a reduction in the availability of psychological therapies for such people. PMID- 12722899 TI - Risks of intravenous cross-contamination infections in the dental office. PMID- 12722901 TI - Transpatellar fixation. PMID- 12722902 TI - Medical ethics. PMID- 12722903 TI - Hip challenges. PMID- 12722904 TI - Universal care may be a solution. PMID- 12722900 TI - Pathogenesis of postoperative oral surgical pain. AB - Pain is a major postoperative symptom in many oral surgical procedures. It is a complex and variable phenomenon that can be influenced by many factors. Good management of oral surgical pain requires a detailed understanding of the pathogenesis of surgical pain. This article aims at reviewing postoperative pain from a broad perspective by looking into the nociception, neuroanatomy, neurophysiology, and neuropharmacology of pain. Therapeutic recommendations are made after reviewing the evidence from the literature for maximizing the efficacy of pain management techniques for oral surgical pain. PMID- 12722905 TI - Radiologic case study. Partial tear of the distal biceps tendon with mass-like bicipitoradial bursitis and associated hyperostosis of the radial tuberosity. PMID- 12722906 TI - Cages are the preferred treatment for large acetabular defects. PMID- 12722907 TI - Treatment of large acetabular defects with jumbo cups. PMID- 12722908 TI - The running near-near-far-far stitch for closure of fasciotomies and other large wounds. AB - The running near-near-far-far stitch is useful for the closure of large wounds such as those created by fasciotomy. The far limb of the suture can be placed outside the injury zone and the near limb helps approximate the wound edges. The running nature of the suture balances the tension throughout the wound and allows the suture to be tightened once swelling has receded. PMID- 12722909 TI - Comparison of surgical outcome between bursal and articular partial thickness rotator cuff tears. AB - Surgical outcomes after arthroscopic subacromial decompression and debridement in bursal and articular partial thickness rotator cuff tears with a tear depth of < 50% were compared. Twenty-four articular and 13 bursal partial thickness rotator cuff tears were evaluated for pain relief and functional recovery. At 6 months postoperatively, the average pain score decreased from 6.2 to 1.7 in patients with articular tears and from 7.1 to 0.9 in patients with bursal tears. Although pain relief and functional recovery were excellent in both groups, the results were better in patients with bursal partial thickness rotator cuff tears at 6 months postoperatively. PMID- 12722910 TI - Repair of full-thickness cartilage defects in rabbit knees with free periosteal graft preincubated with transforming growth factor. AB - This study compared different concentrations of transforming growth factor-beta 1 (TGF-beta1) used for short-term preincubation in vitro of large periosteal explants to determine the effect of chondrogenesis and the fate of repair cartilage over time. Eighty-two rabbits were divided into four groups: group A, non-grafted; group B, non-incubated grafted; group C, 100 ng/mL recombinant human (rh) TGF-beta1 preincubated grafted; and group D, 20 ng/mL rhTGF-beta1 preincubated grafted. Rabbits from each group were sacrificed at intervals between 2 and 24 weeks. Histologic slides were stained with safranin O and were scored based on a subjective scoring system. Group A healed with non cartilaginous material. Group B healed with hyaline cartilage-like material with progressive thinning of this regenerated layer; at 24 weeks, this layer was fibrous tissue. Group C enhanced repair with hyaline cartilage-like material but accelerated early degeneration and osteophyte formation; the cartilage became fibrous tissue at 24 weeks. Group D did not enhance cartilaginous repair. At 24 weeks, all groups had the same result. The 100 ng/mL rhTGF-beta1 preincubation in vitro with periosteum enhanced early osteochondral repair but did not show prolonged durability. Clinical application of this growth factor necessitates further study. PMID- 12722911 TI - Coccygodynia: treatment. AB - This article presents a retrospective review of the treatment of coccygodynia. The past 5 years of conservative treatment for coccygodynia were reviewed, including local injection. The results were evaluated. Retrospectively, the past 20 years of surgical treatment for coccygodynia were reviewed and the clinical results were evaluated. Twenty-four patients were treated with local injection and 15 patients were treated with coccygectomy. Local injection was successful in 78% of patients. Coccygectomy was successful in 87% of patients. The results of conservative treatment with local injection for coccygodynia appear to be successful. However, no other historical literature exists to compare these results. The results of coccygectomy for coccygodynia were also highly successful, and the success rate compares favorably to previous historical data in the literature. PMID- 12722912 TI - Complications related to postoperative casting after surgical treatment of subluxed/dislocated hips in patients with cerebral palsy. AB - Three hundred sixteen subluxed/dislocated hips (222 patients) underwent upper femoral osteotomy. Of these, 286 (90.5%) hips were casted (average patient age: 8.9 years) and 30 (9.5%) were not (average patient age: 13.6 years). Average follow-up was 4.7 years. Complications in the casted/noncasted groups (per hip) were: 43/0 (15%/0%) skin sores; 11/1 (3.8%/3.3%) wound infections; 6/0 (2.1%/0%) instrumentation failures; 22/1 (7.7%/3.3%) reoperations; and 13/1 (4.5%/3.3%) rehospitalizations. Differences between the groups were not statistically significant. Casted patients were younger and more neurologically involved. Casting is useful to ensure healing of osteotomies, prevent instrumentation failure and injury to the operated legs, and allow for ease of handling. Complications that occurred were managed and had no long-term sequelae. PMID- 12722913 TI - Intraoperative hemostasis during spinal reconstructive procedures. AB - In a consecutive sample of patients undergoing similar spinal reconstructive procedures in the thoracolumbar region, hemostatic effectiveness of a collagen based composite (CoStasis; Cohesion Technologies, Palo Alto, Calif) n = 10) was compared with standard hemostatic methods (manual compression) (n = 9). All patients treated with CoStasis at the operative site(s) demonstrated immediate hemostasis,whereas all cases treated with gauze and sponges showed protracted bleeding (P < .01). The average perioperative blood loss was approximately twice as great among control patients compared to CoStasis patients (1322 mL versus 685 mL; P = .02). Total transfusion requirements and operative duration also were less among CoStasis patients. These findings suggest CoStasis may provide effective hemostasis in spinal reconstructive procedures such as instrumented fusion where blood loss is excessive. PMID- 12722914 TI - Diabetic neuroarthropathy (Charcot joints): the importance of recognizing chronic sensory deficits in the treatment of acute foot and ankle fractures in diabetic patients. AB - Patients with diabetic neuropathy are at a higher risk of developing complications, especially Charcot arthropathy. Early diagnosis and intervention is the key to optimizing outcome. Therefore, diabetic patients with a lower extremity injury should be screened with sensory testing using a 5.07 monofilament. PMID- 12722916 TI - Obturator internus tendinitis as a source of chronic hip pain. PMID- 12722915 TI - Failed-back syndrome as a complication of epidural free fat grafts. PMID- 12722917 TI - Uncommon presentation of a medial meniscal tear. PMID- 12722918 TI - Intraneural ganglion of the ulnar nerve at the wrist. PMID- 12722919 TI - Biomechanics of soft-tissue interference screw fixation for anterior cruciate ligament reconstruction. PMID- 12722920 TI - Role of Chlamydia pneumoniae-infected macrophages in atherosclerosis developments of the carotid artery. AB - Chlamydia pneumoniae (C. pneumoniae) infection has been recently accepted as an important cause of atherosclerosis. However, the precise mechanisms remain unclear. The present study was aimed to clarify the distribution link among C. pneumoniae, chlamydial HSP 60, and activated macrophages. Atheromatous carotid plaques were obtained from 40 consecutive carotid endarterectomies (CEA). The specimens were prepared for HE and elastica-van Gieson staining. Parallel sections were stained immunocytochemically with monoclonal antibodies for a C. pneumoniae-specific antigen, chlamydial HSP 60, activated macrophages, and smooth muscle cells. Immunoreactivity for the C. pneumoniae-specific antigen was observed within the endothelial cells, activated macrophages, and smooth muscle cells in 36 of 40 specimens (90%). Chlamydial HSP 60 was found in all specimens positive for the C. pneumoniae-specific antigen, and mainly co-localized with the C. pneumoniae-specific antigen within the activated macrophages. The present results suggest that C. pneumoniae is a key microbial organ that causes atheroma developments in the carotid artery. Chlamydia pneumoniae-infected macrophages may come into the arterial intima and mediate inflammatory and autoimmune processes through the production of chlamydial HSP 60, leading to atherosclerosis. PMID- 12722921 TI - Immunohistochemical study of the expression of cytokines and nitric oxide synthases in brains of patients with dementia with Lewy bodies. AB - Regional expression of cytokines (IL-1alpha, TNF-alpha), inducible nitric oxide synthase (iNOS) and neuronal NOS (nNOS) was immunohistochemically investigated in the brains of patients with dementia with Lewy bodies (DLB), compared with those of patients with Alzheimer's disease (AD) and non-demented elderly persons. It has been reported that inflammatory responses by cytokines and oxygen free radicals such as nitric oxide (NO) are associated with damaged neurons, degenerative neurites or amyloid deposits in AD brains. In the present study, overexpression of IL-1alpha, TNF-alpha and iNOS was demonstrated in the amygdala, hippocampus, entorhinal and insular cortices of DLB brains, which are pathologically the most vulnerable regions in DLB brains as well as AD brains. In addition, some Lewy body (LB)-bearing neurons were involved by the processes of IL-1alpha- and TNF-alpha-positive microglia, and most extracellular LB were associated with the processes of TNF-alpha- and iNOS-positive astroglia. Glial involvement was also found around neuritic plaques and extracellular neurofibrillary tangles. In contrast, the expression of nNOS was reduced in the amygdala of DLB brains showing severe Lewy pathology. These findings suggest that cytokines and NO are significantly implicated in neuronal damage and death including LB formation in DLB brains. PMID- 12722922 TI - Expression of presenilin 1 and synapse-related proteins during postnatal development is not different between accelerated senescence-prone and -resistant mice. AB - SAMP1TA/Ngs is an inbred strain of senescence-accelerated mice in which there is delayed development of cognitive functions and dendritic spine formation compared with normal control SAMR1TA//Ngs mice. It is hypothesized that abnormalities might be in the postnatal expression of synapse-related proteins in SAMP1TA/Ngs mice. Quantitative western blot analyses showed that the postnatal developmental changes in the expression of synaptophysin, post-synaptic density protein 95 and presenilin 1 in the cerebrum were similar between SAMP1TA/Ngs and SAMR1TA//Ngs mice. Therefore, the expression of synapse-related proteins was not disturbed in SAMP1TA/ Ngs mice regardless of reported abnormal numbers of dendritic spines during postnatal development. Immunohistochemical studies showed that the expression of synaptophysin in the neuropil increased postnatally with development in the same way in SAMP1TA/Ngs and SAMR1TA//Ngs mice. Presenilin 1 expression was relatively high at age 5 days in the neuropil of the cerebral cortex and decreased with postnatal development in the same way in SAMP1TA/Ngs and SAMR1TA//Ngs mice. At age 5 days the distribution of presenilin 1 was similar to the distribution of synaptophysin in that there were two separate immunoreactive patterns: a subpial band and patches in the middle layers reminiscent of barrels. These findings suggest that presenilin 1 is transiently expressed in the neuropil to induce synaptogenesis, and then its expression decreases overall. PMID- 12722923 TI - Neuropathological study of the role of mast cells and histamine-positive neurons in selective vulnerability of the thalamus and inferior colliculus in thiamine deficient encephalopathy. AB - The purpose of the present study was to examine the role of histamine in the pathogenesis of experimental thiamine-deficient encephalopathy. By studying sagittal serial sections the authors were able to examine the topographical relationship between histamine-positive neurons and fibers, the number of mast cells, and localized lesions in the thalamus (TH) and inferior colliculus (IC). Adult rats were given a thiamine-deficient diet and pyrithiamine was given intraperitoneally (30 microg/100 g bodyweight per day), and the distribution of vulnerable regions and petechial bleeding was histologically examined by reconstruction of the sagittal serial sections. The distribution of mast cells and histamine-positive neurons and fibers was examined immunohistochemically in control rats, and compared between the vulnerable and non-vulnerable regions of the TH and tectum. Changes in the aforementioned measures during the thiamine deficient state were also examined. The blood-brain barrier was examined using antibodies against rat endothelial barrier antigen (EBA) and albumin. The density of histamine-positive fibers in the vulnerable regions of the TH and IC was very low and not different from the non-vulnerable regions, and the number of mast cells was significantly higher in the lateral portion of the TH than the medial portion of the TH. The numbers of mast cells increased on days 7-10 after the start of the experiment, and significantly decreased on days 14-21. Histamine positive neurons and fibers in the TH and IC also had the same changes. Bleeding of the IC occurred exclusively around arteries, and perivenous bleeding was absent. Albumin exudation and suppression of EBA expression of capillaries were found in the spongy lesions of the TH and IC. The role of histamine in selective vulnerability of the TH and IC in experimental thiamine-deficient encephalopathy was not supported. Findings in the present study suggest that the spongy change is a primary event, and vascular changes are secondary. PMID- 12722924 TI - Glyceraldehyde 3-phosphate dehydrogenase and endothelin-1 immunoreactivity is associated with cerebral white matter damage in dentatorubral-pallidoluysian atrophy. AB - DRPLA is a rare neurodegenerative disorder caused by CAG triplet elongation on chromosome 12p. In addition to neurodegeneration of both the dentatorubral and pallidoluysian systems, there is cerebral white matter damage, especially in older cases. Intracellular accumulation of DRPLA protein is widespread in the central nervous system, and DRPLA protein has been shown to immobilize glyceraldehyde 3-phosphate dehydrogenase (GAPDH), which regulates glycolysis and controls mRNA of tissue-type plasminogen activator (tPA) in tissue restoration. However, little is known about the pathogenesis regarding the formation of cerebral white matter damage in DRPLA. Therefore, the pathology of this damage was investigated by examining markers of glycolysis and related processes. Nine clinically and pathologically confirmed DRPLA cases were used in the present study. CAG triplet elongation on chromosome 12p was confirmed in all cases where tissue was available for genotyping (seven cases). PAS and immunohistochemistry with antibodies to GFAP, GAPDH and endothelin-1 were used to demonstrate astrocytosis. The polysaccharides storage state with PAS-positive astrocytes was detected in seven cases. GAPDH- and endothelin-1-positive endothelium and astrocytes were observed in two cases with GFAP-positivity. Based on the biochemical process together with the present results, GAPDH and endothelin-1 immunoreactivity is associated with this damage and the mismetabolism of polysaccharides caused by CAG triplet elongation on chromosome 12p may contribute to the formation of the cerebral white matter damage in DRPLA. PMID- 12722925 TI - Neuropathological study of C57BL/6Akita mouse, type 2 diabetic model: enhanced expression of alphaB-crystallin in oligodendrocytes. AB - The structure of the central and peripheral nervous systems was studied. in the C57BL/6Akita (Akita) mouse, a non-obese type 2 diabetes model characterized by early onset, autosomal dominant inheritance and a mutation of the insulin 2 gene. Usual neuropathological examinations showed no remarkable abnormalities in the brain, spinal cord or sciatic nerve of Akita mice up to 48 weeks of age. However, immunohistochemical examination revealed that expression of alphaB-crystallin was enhanced in oligodendrocytes in the cerebral white matter, especially in the corpus callosum, after 32 weeks of age. The oligodendrocytes were not positive for ubiquitin and HSP25. It is suggested that long-standing hyperglycemia might stress the CNS and thus enhance the expression of alphaB-crystallin in oligodendrocytes. PMID- 12722926 TI - Lack of amyloid plaque formation in the central nervous system of a patient with Werner syndrome. AB - Werner syndrome (WS) is an autosomal recessive disorder associated with accelerated aging. It is well documented on systemic aging but it is unclear whether the brain with WS shows accelerated aging. A 55-year-old patient with WS was studied and it was found that a deletion mutation of exon 26 of the WRN gene was not associated with CNS pathology, such as amyloid plaques or NFT. Furthermore, additional genetic analysis showed an apolipoprotein E genotype of epsilon3/epsilon3 that did not play either an accelerating or inhibitory action on' amyloid deposition. Therefore, based on the genetic and neuropathological analysis, it was observed that the WS-associated aging seen in many organs did not extend to the CNS. PMID- 12722927 TI - Lung carcinoma metastasis presenting as a pineal region tumor. AB - An autopsy case is reported of pineal metastasis from lung adenocarcinoma, which is a rare manifestation of the disease. A 75-year-old man who had been found to have a lesion in the lung by chest CT 2 years previously, became aware of head heaviness and then suffered consciousness disturbance. Brain MRI revealed a solitary mass in the pineal region with hydrocephalus. At autopsy a midsagittal section of the brain disclosed a well-circumscribed mass consisting of epithelial cells occupying the third ventricle. Although it should be recognized that such metastasis is very rare, the present case provides further information that might be useful for diagnosis. PMID- 12722928 TI - Fundamentals of the morphometric approach in neuropathology. AB - The purposes of neuropathological examination are diagnosis and discovery of new findings. Quantitative examination closely relates to these aims. To fulfil these aims, neuropathologists should understand well the strengths and weaknesses of the power of observation. The power of observation in humans is quite strong in differentiation of shapes, but weak in counting numbers and measuring size, length and width. Quantitative examination is a help in observation, and the fruits of the examination are generalization, numerization and schematization of the neuropathological findings, and analogy of neuronal function. PMID- 12722929 TI - Tissue culture methods to study neurological disorders: establishment of immortalized Schwann cells from murine disease models. AB - Previously, the authors have established spontaneously immortalized cell lines from long-term cultures of normal adult mouse Schwann cells. Establishment of such Schwann cell lines derived from murine disease models may greatly facilitate studies of the cellular mechanisms of their peripheral nervous system lesions in the relevant diseases. Recently, the authors have established immortalized Schwann cell lines derived from Niemann-Pick disease type C mice (NPC; spm/spm) and globoid cell leukodystrophy mice (twitcher). In the present study, long-term cultures were maintained of Schwann cells derived from dorsal root ganglia and consecutive peripheral nerves of another NPC mouse (npc(nih)/npc(nih), npc(nih)/+), myelin P0 protein-deficient mice (P0-/-, P0+/-) with their wild-type littermates (P0+/+), and neurofibromatosis type 1 gene (NF1)-deficient mice (Nf1(FCr)/+) for 8-10 months, and immortalized cell lines from all these animals established spontaneously. These cell lines had spindle-shaped Schwann cell morphology and distinct Schwann cell phenotypes and retained genomic and biochemical abnormalities, sufficiently representing the in vivo pathological features of the mutant mice. These immortalized Schwann cell lines can be useful in studies of nervous system lesions in these mutant mice and relevant human disorders. PMID- 12722930 TI - Checkpoints and pitfalls in the experimental neuropathology of circulatory disturbance. AB - In neural tissue injury many pathological processes are common to different neurological disorders, including cerebral ischemia. Because ischemia has a fundamentally simple impact on neural tissue, good laboratory modeling can help improve the general understanding of the neuropathological processes involved. Summarized here are some basic principles that should be followed to ensure that cerebral ischemia studies are reproducible and informative: (i) selection of an appropriate model of cerebral ischemia in an appropriate species (although rodents are widely used for genomic studies, the use of larger animals, with brain structures macroscopically similar to those of humans, is appropriate for many studies, e.g. of white matter lesions or the pathophysiology of cerebral edema); (ii) correct maintenance of physiological parameters, including body temperature, systemic blood pressure, and blood gas tensions, under appropriate general anesthesia; (iii) selection of an appropriate method of cerebral blood flow (CBF) monitoring (decisions include whether or not the experiment requires real-time monitoring, in vivo measurement, and CBF mapping); (iv) appropriate timing of drug application in therapeutic studies (many drugs that are effective when given immediately after a short period of ischemia are ineffective in clinical trials, probably because of longer periods of ischemia and delayed drug delivery in clinical settings); and (v) multiparametric evaluation of therapeutic effect (with the recent increase in diagnosis of cases of mild stroke, measurement of mortality and infarct size have proven to be insufficient for the evaluation of therapeutic effect). Use of mild ischemia models and batteries of neurological tests for individual neurological functions, such as motor, somatosensory, and visual function, are becoming important in experimental ischemia research. In histological evaluation, assessment of the extent of both selective neuronal loss and the infarct will become mandatory. Regional analysis of each brain structure and coordination of the results with the apparent neurological dysfunction is a promising approach. PMID- 12722931 TI - Genotype-phenotype correlation: familial Parkinson disease. AB - Kindreds with Mendelian inheritance of Parkinson disease (PD) have been known since a long time ago. Nine loci have been mapped in familial PD by linkage study and four causative genes have been cloned. This paper discusses Park 1 and Park 2, the identification of which has brought about many advances in the studies on pathomechanism of PD. Investigations of these genes in familial PD have expanded their clinical and pathological phenotypes. However, to clarify the effect of mutations on these phenotypes, additional post-mortem neuropathological studies are required. PMID- 12722932 TI - Genetic alterations of human brain tumors as molecular prognostic factors. AB - Despite a number of basic and clinical studies, it is still very difficult to improve the prognosis of patients with high-grade astrocytoma. However, the recent success of procarbazine, N-(2-chloroethyl)-N'-cyclohexyl-N-nitrosourea and vincristine (PCV) chemotherapy for oligodendrogliomas that have lost chromosomes 1p and 19q has encouraged the authors to evaluate the biological behavior of brain tumors by means of genetic analysis. Both the disorders of the p53/MDM2/p14(ARF) and the p16(INK4a)/RB signaling pathways have been found to play an essential role in tumorigenesis of various brain tumors. Herein, the authors summarize the genetic alterations of brain tumors by mainly focusing on two pathways that appear to affect significantly the patient prognosis. PMID- 12722933 TI - Binaural beat induced theta EEG activity and hypnotic susceptibility: contradictory results and technical considerations. AB - The present study offered a constructive replication of an earlier study which demonstrated significant increases in theta EEG activity following theta binaural beat (BB) entrainment training and significant increases in hypnotic susceptibility. This study improved upon the earlier small-sample, multiple baseline investigation by employing a larger sample, by utilizing a double-blind, repeated-measures group experimental design, by investigating only low and moderate susceptible participants, and by providing 4 hours of binaural beat training. With these design improvements, results were not supportive of the specific efficacy of the theta binaural beat training employed in this study in either increasing frontal theta EEG activity or in increasing hypnotic susceptibility. Statistical power analyses indicated the theta binaural beat training to be a very low power phenomenon on theta EEG activity. Furthermore, we found no significant relationship between frontal theta power and hypnotizability, although the more hypnotizable participants showed significantly greater increases in hypnotizability than the less hypnotizables. Results are discussed within the context of participant selection and classification factors, technical considerations in the presentation of TBB training, and theta blocking. PMID- 12722934 TI - Working with Dissociative Fugue in a general psychotherapy practice: a cautionary tale. AB - Dissociative Fugue is a somewhat rare condition that therapists may see only once or twice over the course of a professional career. A brief review of the uses of hypnosis in the treatment of Dissociative Fugue is followed by a presentation of the case of a 51-year-old man who presented with the clinical picture of Dissociative Fugue State and who experienced complete amnesia for the time prior to the fugue state. This article focuses on the pitfalls that the psychotherapist in a general practice may face when working with such a patient and offers specific recommendations and scripts that may be useful in proceeding with treatment. PMID- 12722935 TI - Antaeus and androgogy: negotiating paradigm exhaustion and pursuing professional growth in clinical practice. PMID- 12722936 TI - Can medical hypnosis accelerate post-surgical wound healing? Results of a clinical trial. AB - Although medical hypnosis has a long history of myriad functional applications (pain reduction, procedural preparation etc.), it has been little tested for site specific effects on physical healing per se. In this randomized controlled trial, we compared the relative efficacy of an adjunctive hypnotic intervention, supportive attention, and usual care only on early post-surgical wound healing. Eighteen healthy women presenting consecutively for medically recommended reduction mammaplasty at an ambulatory surgery practice underwent the same surgical protocol and postoperative care following preoperative randomization (n = 6 each) to one of the three treatment conditions: usual care, 8 adjunctive supportive attention sessions, or 8 adjunctive hypnosis sessions targeting accelerated wound healing. The primary outcome data of interest were objective, observational measures of incision healing made at 1,7 weeks postoperatively by medical staff blind to the participants' group assignments. Data included clinical exams and digitized photographs that were scored using a wound assessment inventory (WAI). Secondary outcome measures included the participants' subjectively rated pain, perceived incision healing (VAS Scales), and baseline and post-surgical functional health status (SF-36). Analysis of variance showed the hypnosis group's objectively observed wound healing to be significantly greater than the other two groups', p < .001, through 7 postoperative weeks; standard care controls showed the smallest degree of healing. In addition, at both the 1 and 7 week post-surgical observation intervals, one-way analyses showed the hypnosis group to be significantly more healed than the usual care controls, p < 0.02. The mean scores of the subjective assessments of postoperative pain, incision healing and functional recovery trended similarly. Results of this preliminary trial indicate that use of a targeted hypnotic intervention can accelerate postoperative wound healing and suggest that further tests of using hypnosis to augment physical healing are warranted. PMID- 12722937 TI - Cleaning up the river: a metaphor for functional digestive disorders. AB - The irritable bowel syndrome is characterized by abdominal pain, altered bowel habits and various other digestive symptoms. Emotional factors are important in some patients. I describe here a metaphor which links the altered motility of the digestive system to the emotional contents it may embody. A metaphor of a river is used to evoke both a smooth, coordinated flow through the normal digestive tract and a normal flow in the management of the patient's emotions. The possibility that some blockage has occurred in the river, resulting in perturbation of the normal flow is then suggested to the patient. This is followed by a suggestion for the patient to clear the blockage. This approach may lead patients to work on the emotional components of their symptoms, resulting in their subsequent resolution. I have used this approach with several patients and it proved very effective. This paper demonstrates the use of this metaphor in one of the patients. PMID- 12722938 TI - Poly(ethyleneimine)-immobilized-cloth enzyme immunoassay for the detection of Salmonella lipopolysaccharide. AB - Poly(ethyleneimine) (PEI) was immobilized on non-woven polyester cloth and examined for application on a simple, rapid and economical "cloth enzyme immunoassay (CEIA)" which was developed originally as polymyxin-CEIA for the detection of Salmonella lipopolysaccharide (LPS). PEI-cloth regardless of the PEI molecular weight, but with the amine group contents of 0.1 to approximately 0.35 meq/g immobilized either in a physisorption-like or chemisorption-like manner, adsorbed LPS rapidly, preferentially and effectively. The captured LPS was then able to be detected qualitatively and quantitatively as an antigen by enzyme immunoassay. PEI-CEIA had a detection limit for Salmonella LPS of 10 ng/ml, which was equivalent to 1.6 x 10(5) cell/ml and was ten times more sensitive than polymyxin-CEIA. It was possible to detect Salmonella LPS in the presence of a 100 fold excess of E. coli LPS. PEI-CEIA was found to be more sensitive and much easier to carry out than polymyxin-CEIA but had the same advantages as polymyxin CEIA. PMID- 12722939 TI - Disregulated expression of the Th2 cytokine gene in patients with intraoral squamous cell carcinoma. AB - It has been seen that advanced stage oral squamous cell carcinoma is associated with impaired T-cell function and higher antibody response. In order to find out if such immune disregulation is associated with alteration of T-helper (Th) type CD4+ T-cell phenotype leading to altered cytokine production, we studied the Th like cytokine profile in 35 oral squamous cell carcinoma patients and 21 normal controls. Concomitant expression of both Th1 and Th2 cytokine genes was studied by reverse transcription and Polymerase Chain Reaction (PCR) based amplification (RT-PCR) of mRNA extracted from freshly isolated peripheral blood mononuclear cells (PBMC) using specific primers for Interferon (IFN)-gamma, Interleukin (IL) 2, IL-4 and IL-10. Almost 63% of oral cancer patients showed polarization of a Th like cytokine response as compared to 33% of the normal controls while 66.6% of normal controls showed a predominantly non-polarized Th0 response. Expression of IFN-gamma and IL-2 genes was more commonly seen in the early stage of the disease (p < 0.02) whereas majority of advanced stage tumours was associated with enhanced expression of IL-4 and IL-10 but not IFN-gamma and IL-2 genes. Patients with lymphnode metastases and poorly differentiated tumours expressed IL-4 and IL 10 more frequently with concomitant suppression of IFN-gamma and IL-2 genes. It seems therefore, that the development of oral squamous cell carcinoma leads to polarization of cytokine gene expression that is skewed towards the Th1-like response in the early stage. However, increasing tumour load and lymphnode invasion suppresses Th1 cytokine genes, thus skewing it toward a Th2-like cytokine response. PMID- 12722941 TI - Carvedilol modulates in-vitro granulocyte-macrophage colony-stimulating factor induced interleukin-10 production in U937 cells and human monocytes. AB - Both granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-10 (IL-10) are important mediators regulating inflammatory responses. Inflammatory processes have an important role in atherogenesis. In this paper, the effects of carvedilol on GM-CSF-induced IL-10 production were examined on human monocytic cell line, U937, and purified human monocytes. First, we showed that one-time carvedilol pretreatment at concentrations 0.3-10 microM dose-dependently inhibited GM-CSF-induced IL-10 production in U937 cells. In addition, we found carvedilol to be non-cytotoxic at concentrations equal to or less than 10 microM. However, at concentrations higher than 10 microM, carvedilol induced programmed cell death in U937 cells. The inhibition of GM-CSF-induced IL-10 production by carvedilol was also observed at the expression of mRNA. Furthermore, the inhibition of IL-10 production was demonstrated in GM-CSF-activated purified human peripheral blood monocytes. Finally, long-term carvedilol pretreatment of U937 cells up to 2 months at concentrations of 1.0 microM mildly enhanced the IL 10 production. Our observations that carvedilol modulated GM-CSF-induced IL-10 production may have some implication in understanding the broad-spectrum effects of carvedilol in regulating inflammatory reactions. PMID- 12722940 TI - Cross-binding between Plasmodium falciparum CTL epitopes and HLA class I molecules. AB - Plasmodium falciparum CTL epitope minigenes containing HLA-A2 and HLA-B7 subtype supermotifs were cloned into a plasmid expression vector; this expression was measured in eight human HLA class I molecule specific cell lines. Three assays for in vitro antigen presentation analysis were developed to examine the cross binding between CTL epitopes and HLA class I molecules, including cell surface peptide-MHC class I binding assay, binding stabilization assay and MHC class I assembling assay. The results demonstrated that the HLA-B51 restricted CTL epitope of Plasmodium falciparum could be presented by other HLA class I molecules; however, no other presentation was found for HLA-A2.1 CTL epitope. This work suggests the possibility for improved vaccine-coverage rates by development of a CTL vaccine which contains epitopes capable of cross-binding among different MHC class I alleles. PMID- 12722942 TI - Tyrosine mutation in CD3epsilon-ITAM blocked T lymphocyte apoptosis mediated by CD3epsilon. AB - Anti-CD3epsilon monoclonal antibody induces programmed cell death of thymocytes and accelerates activation-induced cell death (AICD) by apoptosis of matured or transformed T lymphocytes. However, the underlying molecular mechanism of this phenomenon is unclear. Therefore, we produced a chimera protein (termed CD8epsilon by fusing the extracellular and transmembrane domains of human CD8alpha to the intracellular domain of mouse CD3epsilon and expressed in CD8- Jurkat T cells. Stable cell lines of mutants expressing the motifs of Y170F, Y181F, and Y170F/Y181F in the CD3epsilon-ITAM were established. Experiments showed that apoptosis could be induced only in the T Jurkat cells with intact CD3epsilon intracellular domain, but not in the cells with the mutant CD8epsilon when stimulated with anti-CD8alpha monoclonal antibody. This finding indicated that a single tyrosine mutation in CD3epsilon-ITAM blocked the signal transduction, causing the cell death by apoptosis when stimulated by CD8epsilon molecule. During the apoptotic process, we showed that expressions of CD95, CD95L and Nur77 were enhanced in stimulated TJK cells but not in control cells. In addition, the high expression of Nur77 kept pace with the onset of apoptosis of T cells mediated by CD8epsilon. We further showed that 3'-phosphatidylinositol kinase (PI3K) were not only enhanced during T cell activation, but also in the AICD process. The results suggest that PI3K/Akt is not only a cell proliferation signal, but also a potential apoptosis regulator in T lymphocytes. PMID- 12722943 TI - The H-Y response in mid-gestation and long after delivery in mice primed before pregnancy. AB - The mechanisms underlying maternal tolerance of the semi-allogeneic fetus are not completely understood. The maternal immune system's response to the male antigen, H-Y is an example of the conflicting evidence that both supports and refutes the idea that the immune system in pregnant females is fundamentally different from that in non-pregnant females. Although multiple pregnancies may inactivate H-Y specific T cells, the immune system of the pregnant female can also generate a cytotoxic response to this antigen. To help understand this apparent conflict, we immunized female mice against H-Y with male spleen cells before pregnancy and examined the subsequent anti H-Y response during mid-pregnancy. The pregnant mice studied were able to mount cytotoxic immune responses to H-Y that were equivalent to those generated in their non-pregnant counterparts. Moreover the experience of pregnancy did not impair the ability to maintain immunologic memory to H-Y. The data support the idea that pregnancy does not violate general rules of antigen specific immunity, even if the antigen is expressed on the fetus. PMID- 12722944 TI - Better survival of Helicobacter pylori infected patients with early gastric cancer is related to a higher level of Thomsen-Friedenreich antigen-specific antibodies. AB - The survival of patients with histologically verified gastric carcinoma at stage I (n = 44) and stage II (n = 43) was analysed by the Kaplan-Meier method depending on H. pylori serological status and a level of IgG and IgM antibody to tumor-associated Thomson-Friedenreich antigen (T Ag). In cancer patients at stage I, significantly better survival for H. pylori seropositive patients was observed compared to H. pylori seronegative patients (median SE survival time: 60.0 +/- 3.8 mths and 37.0 +/- 7.8 mths, respectively; P < 0.0004, log-rank test). Patients with higher level of T Ag-specific IgG antibody (strong responders) showed significantly and dramatically better (P < 0.00001) survival rate than weak responders. However, an association of better survival with a higher level of anti-T antibody level was limited to the H. pylori seropositive patients exclusively (P < 0.00001) with no difference for H. pylori seronegative group of patients. The level of IgM anti-T Ag antibody was not significantly related to the survival of patients at both stages of the disease, though better survival was noted in H. pylori seropositive IgM strong responders at approximately 40-60 months of observation. Statistically insignificant associations between survival and H. pylori status or anti-T antibody levels were also observed in a group of gastric cancer patients at stage II. In summary, the survival of patients with early gastric cancer (stage I) is significantly better in H. pylori seropositive patients, and this phenomenon may be in part explained by up-regulation of T Ag specific IgG immune response in H. pylori infected individuals. PMID- 12722945 TI - Comparative analysis of mononuclear cell surface markers in atopic processes--a preliminary study. AB - Atopic disorders are driven by the Th2 cell subset. We have determined the expression of costimulatory molecules and cell surface markers on peripheral CD4+ T cells and antigen presenting cells, in different atopic diseases, and we have also tried to correlate the expression of these markers with the severity of the disease. Cells from patients with atopic and contact dermatitis, mild or severe asthma, and symptomatic and non-symptomatic atopic rhinitis were analyzed by flow cytometry. Our results showed that CD30, CD124, and CD152 expression on CD4+ T cells was significantly higher in atopic dermatitis than in contact dermatitis patients (p < 0.05). It was interesting to observe that the cell surface expression of CD80 in T and B cells from atopic dermatitis patients was not enhanced as opposed to the other atopic diseases we analyzed. Our results suggest that there are differences in the immune mechanisms involved in the different atopic diseases, and that expression of CD30 in CD4+ T cells might be a marker of disease activity in atopic dermatitis. PMID- 12722946 TI - Bystander T cells participate in specific response to cockroach antigen (CR) in vitro. AB - Allergic reactions due to whole body, body parts and fecal products of cockroach (CR) are characterized by inflammatory reaction that may lead to symptoms of rhinitis or asthma in atopic individuals. Although the majority of T cells at the site of CR hypersensitivity are not antigen specific, the cellular subset and cytokine receptors that participate and control the outcome of the reaction have not been fully studied. In this study, we have used fluorescent activated cell sorter (FACS) analysis to characterize the activation marker and cytokine profile of antigen specific and bystander T cells after in vitro stimulation of peripheral blood lymphocytes with whole body extract of CR antigen. There was significant enhancement of CD69 on blast and bystander T cells in all atopic subjects compared to non-atopics. Both antigen specific and bystander T cells showed increased expression of HLA-DR, CD25 and CD71 in 9 of 11 atopic patients compared to control. There was also an increase in CD45RA+ and a decrease in CD45RO+ cells following antigen stimulation. These results correlated with the increase in the early apoptotic cells observed in patients as measured by Annexin V stain. Our data revealed that there was no difference in the expression of CD95 in both stimulated and bystander T cells. However, there was enhancement of FasL by CR antigen, suggesting that the increased apoptosis that was observed was probably due to the Fas/FasL interaction. Positive intracellular IL2, IL-4 and IFN-gamma in T cells were observed in only the antigen specific blast cells in 83% of patients studied. These results suggest interplay of memory T cell response, apoptosis, and activated bystander T cells activities in maintaining cellular homeostasis during allergic reaction in cockroach sensitive atopic subjects. PMID- 12722947 TI - Distribution, function, and properties of leptin receptors in the brain. AB - Leptin, a peptide hormone, is implicated in the modulation of food intake and maintenance of energy balance in many vertebrates including humans. It is considered to act via its receptor mainly through several hypothalamic nuclei that play critical roles in the regulation of appetite. This article looks mainly at the functional significance of leptin in rat brain by drawing on published reports of morphological and physiological analyses. Our immunohistochemical observations indicate that the leptin receptor is distributed throughout the brain, including the hypothalamus, and interestingly, is found in the hippocampus and neocortex. Physiological experiments with single living cells isolated from fresh rat hypothalamus clearly demonstrate that leptin has a significant effect on feeding-regulating neurons in the hypothalamus. Studies to date support a role for leptin not only in modulating food intake and appetite in rats and humans, but also in relation to learning and memory processes. PMID- 12722948 TI - Apoptosis and necrosis in health and disease: role of mitochondria. AB - Mitochondria play an important role in both the life and death of cells. Mitochondria are the powerhouse of the cell, providing over 90% of adenosine triphosphate (ATP) consumed by the cell. Mitochondrial energy production, however, is disrupted in various pathological situations leading to cellular Injury. The mechanisms causing the injury are turning out to be more complex than originally expected. For instance, calcium, oxidant chemicals, ischemia/ reperfusion, and a range of other agents promote onset of the mitochondrial permeability transition in mitochondria from liver, heart, and other tissues. Often the consequence of this event is ATP depletion, ion deregulation, mitochondrial and cellular swelling, activation of degradative enzymes, plasma membrane failure, and cell lysis. This is referred to as necrotic cell death. The mitochondrial permeability transition is also involved in apoptotic cell death. In this mode of death, the role of the permeability transition is to release proapoptotic proteins from mitochondria into the cytosol where with the aid of cellular ATP they complete the apoptotic cascade. Therefore, mitochondria contribute to both apoptotic and necrotic death. PMID- 12722949 TI - The apicoplast: a plastid in Plasmodium falciparum and other Apicomplexan parasites. AB - Apicomplexan parasites cause severe diseases such as malaria, toxoplasmosis, and coccidiosis (caused by Plasmodium spp., Toxoplasma, and Eimeria, respectively). These parasites contain a relict plastid-termed "apicoplast"--that originated from the engulfment of an organism of the red algal lineage. The apicoplast is indispensable but its exact role in parasites is unknown. The apicoplast has its own genome and expresses a small number of genes, but the vast majority of the apicoplast proteome is encoded in the nuclear genome. The products of these nuclear genes are posttranslationally targeted to the organelle via the secretory pathway courtesy of a bipartite N-terminal leader sequence. Apicoplasts are nonphotosynthetic but retain other typical plastid functions such as fatty acid, isoprenoid and heme synthesis, and products of these pathways might be exported from the apicoplast for use by the parasite. Apicoplast pathways are essentially prokaryotic and therefore excellent drug targets. Some antibiotics inhibiting these molecular processes are already in chemotherapeutic use, whereas many new drugs will hopefully spring from our growing understanding of this intriguing organelle. PMID- 12722950 TI - Transcriptional regulation of meiosis in budding yeast. AB - Initiation of meiosis in Saccharomyces cerevisiae is regulated by mating type and nutritional conditions that restrict meiosis to diploid cells grown under starvation conditions. Specifically, meiosis occurs in MATa/MATalpha cells shifted to nitrogen depletion media in the absence of glucose and the presence of a nonfermentable carbon source. These conditions lead to the expression and activation of Ime 1, the master regulator of meiosis. IME1 encodes a transcriptional activator recruited to promoters of early meiosis-specific genes by association with the DNA-binding protein, Ume6. Under vegetative growth conditions these genes are silent due to recruitment of the Sin3/Rpd3 histone deacetylase and Isw2 chromatin remodeling complexes by Ume6. Transcription of these meiotic genes occurs following histone acetylation by Gcn5. Expression of the early genes promote entry into the meiotic cycle, as they include genes required for premeiotic DNA synthesis, synapsis of homologous chromosomes, and meiotic recombination. Two of the early meiosis specific genes, a transcriptional activator, Ndt80, and a CDK2 homologue, Ime2, are required for the transcription of middle meiosis-specific genes that are involved with nuclear division and spore formation. Spore maturation depends on late genes whose expression is indirectly dependent on Ime1, Ime2, and Ndt80. Finally, phosphorylation of Imel by Ime2 leads to its degradation, and consequently to shutting down of the meiotic transcriptional cascade. This review is focusing on the regulation of gene expression governing initiation and progression through meiosis. PMID- 12722951 TI - Myosins and cell dynamics in cellular slime molds. AB - Myosin is a mechanochemical transducer and serves as a motor for various motile activities such as cell migration, cytokinesis, maintenance of cell shape, phagocytosis, and morphogenesis. Nonmuscle myosin in vivo does not either stay static at specific subcellular regions or construct highly organized structures, such as sarcomere in skeletal muscle cells. The cellular slime mold Dictyostelium discoideum is an ideal "model organism" for the investigation of cell movement and cytokinesis. The advantages of this organism prompted researchers to carry out pioneering cell biological, biochemical, and molecular genetic studies on myosin II, which resulted in elucidation of many fundamental features of function and regulation of this most abundant molecular motor. Furthermore, recent molecular biological research has revealed that many unconventional myosins play various functions in vivo. In this article, how myosins are organized and regulated in a dynamic manner in Dictyostelium cells is reviewed and discussed. PMID- 12722952 TI - Structure-specific DNA-binding proteins as the foundation for three-dimensional chromatin organization. AB - Any functions of tandem repetitive sequences need proteins that specifically bind to them. Telomere-binding TRF2/MTBP attaches telomeres to the nuclear envelope in interphase due to its rod-domain-like motif. Interphase nuclei organized as a number of sponge-like ruffly round chromosome territories that could be rotated from outside. SAF-A/hnRNP-U and p68-helicase are proteins suitable to do that. Their location in the interchromosome territory space, ATPase domains, and the ability to be bound by satellite DNAs (satDNA) make them part of the wires used to help chromosome territory rotates. In case of active transcription p68 helicase can be involved in the formation of local "gene expression matrices" and due to its satDNA-binding specificity cause the rearrangement of the local chromosome territory. The marks of chromatin rearrangement, which have to be heritable, could be provided by SAF-A/hnRNP-U. During telophase unfolding the proper chromatin arrangement is restored according to these marks. The structural specificity of both proteins to the satDNAs provides a regulative but relatively stable mode of binding. The structural specificity of protein binding could help to find the "magic" centromeric sequence. With future investigations of proteins with the structural specificity of binding during early embryogenesis, when heterochromatin formation goes on, the molecular mechanisms of the "gene gating" hypothesis (Blobel, 1985) will be confirmed. PMID- 12722953 TI - Stinging nettle. PMID- 12722954 TI - Ouch! Missing the point of needlestick prevention. PMID- 12722955 TI - The allergist as consultant. PMID- 12722956 TI - Diet and asthma: has the role of dietary lipids been overlooked in the management of asthma? AB - OBJECTIVE: This article discusses the role of diet in the management of asthma. Readers will gain an understanding of how evolution of the western diet has contributed to increased asthma prevalence and how dietary modification that includes management of dietary lipids may reduce symptoms of asthma. DATA SOURCES: Relevant studies published in English were reviewed. STUDY SELECTION: Medline search to identify peer-reviewed abstracts and journal articles. RESULTS: Asthma and obesity, which often occur together, have increased in prevalence in recent years. Studies suggest adaption of a western diet has not only contributed to obesity, but that increased intake of specific nutrients can cause changes in the frequency and severity of asthma. Increased asthma prevalence has also been proposed to arise from increased exposure to diesel particles or lack of exposure to infectious agents or endotoxins during childhood, generating a biased Th2 immune response, and increased cytokine and leukotriene production. Antagonists directed against these pro-inflammatory mediators include anticytokines and antileukotrienes. A reduction in the levels of inflammatory mediators associated with asthma has also been seen with dietary interventions, such as the administration of oils containing gamma-linolenic acid and eicosapentaenoic acid. CONCLUSIONS: Evidence suggests elevated body mass index and dietary patterns, especially intake of dietary lipids, contribute to symptoms of asthma. Dietary modification may help patients manage their asthma as well as contribute to their overall health. PMID- 12722957 TI - A 16-month-old with persistent vomiting. PMID- 12722958 TI - Bloody diarrhea, fever, and pancytopenia in a patient with active ulcerative colitis. PMID- 12722959 TI - Needle sticks and adverse outcomes in office-based allergy practices. AB - BACKGROUND: In 1984 the first case of needle stick transmitted human immunodeficiency virus was reported. In 1986 Occupational Safety and Health Administration was petitioned by various unions representing health care employees to develop a standard which protects employees from occupational exposure to blood-borne diseases. Congress passed the Needle Stick Safety and Prevention Act. This specifies that "safer medical devices, such as sharps with engineered sharps injury protections and needle-less systems" constitute an effective engineering control, and must be used where feasible. This has been mandated in California as part of the labor code. Blood-borne pathogens of concern in needle stick injuries are human immunodeficiency virus, hepatitis virus B, and hepatitis virus C. OBJECTIVE: The objective of this study was to determine the incidence of accidental needlesticks (ANSs) and disease transmission in the allergy setting. METHODS: A retrospective survey of most California allergy practices and a few large multi-physician allergy practices. We received and used 121 of 400 surveys. RESULTS: Analysis of the survey data showed an overall incidence of 45 ANSs with 7.026 million 26-/27-gauge needles reported. There was zero rate of disease transmission; 6.41 ANSs per million compares favorably with an estimated 267 ANSs per million in the general medical setting. CONCLUSIONS: The rate of ANSs in the allergist's office is 2% that of general medical ANSs. The current "safety" needles have no proven effectiveness. There is no reported disease transmission in the allergist's office setting using existent methods. This solution needs further study before there is generalized implementation of the engineering devices of no proven effectiveness that may in fact increase ANSs. PMID- 12722961 TI - Determination of the incidence of sensitization after penicillin skin testing. AB - BACKGROUND: Concerns for sensitization after penicillin skin testing are a factor in limiting the timing and population for whom this testing is offered. The sensitizing potential of the penicillin skin test has never been studied directly. METHODS: A total of 329 volunteers underwent prick and intradermal skin testing with penicillin G, benzylpenicilloyl-polylysine, and a minor determinant mixture. Those with negative skin testing had repeat testing 4 weeks later. Medical history and antibiotic use were determined by interview, questionnaire, and electronic pharmacy records. RESULTS: Seventy-two of the 329 subjects (22%) reported a history of previous beta-lactam reaction, of which 10 (14%) had a positive initial skin test. Overall, the initial skin test was positive in 23 of 329 (7%). Of the subjects with a negative initial skin test, 239 completed the second test 4 weeks later. Of these, 6 subjects (2.5%, 95% confidence interval 0.5% to 4.5%) converted to a positive skin test. None had taken a beta-lactam antibiotic between the two tests, and none had any previous history of beta lactam reaction. One subject reported having never taken a beta-lactam antibiotic before. In comparison to the 233 subjects who did not convert their skin test, the statistically significant factors favoring sensitization were: female sex (odds ratio [OR] 6.53, P = 0.05), atopy (OR 5.31, P = 0.04), and history of food allergy (OR 6.35, P = 0.02). There was a trend toward more recent penicillin use in the newly sensitized subjects, but this was not statistically significant.. CONCLUSION: Penicillin skin testing may sensitize a small number of individuals to penicillin. PMID- 12722960 TI - Inpatient consultation of allergy/immunology in a tertiary care setting. AB - BACKGROUND: Few studies examine the referral patterns for allergy/immunology (A/I) inpatient consultation. OBJECTIVE: The purpose of this study was to examine the primary reason and trends for A/I inpatient consultation to improve fellowship training. METHODS: We performed a retrospective chart review of all inpatient A/I consults from July 1, 1987 to June 30, 2001 to determine the primary reason for consultation. We also reviewed trends in the total admissions and the average daily patient load compared with A/I consultation. RESULTS: A total of 1,284 A/I inpatient consults were reviewed. Thirty-six percent (460 of 1,284) of inpatient consults were for evaluation of adverse drug reactions, 21% (270 of 1,284) asthma, 21% (272 of 1,284) miscellaneous reasons, 8% (109 of 1,284) possible immunodeficiency, 7% (93 of 1,284) angioedema/urticaria, and 6% (80 of 1,284) anaphylaxis. Our results demonstrated a fall in inpatient consults that correlated with a similar fall in total hospital admissions. The ratio of A/I inpatient consults to total admissions remained constant. Additionally, the ratio of A/I consults to average daily patient load increased over the study period. There was a decrease in asthma and adverse drug reaction consults, whereas immunodeficiency and anaphylaxis referrals remained stable. CONCLUSIONS: Identifying the most common reasons for inpatient consultation provides a guide for the education of A/I fellows and primary care residents. Inpatient consultation continues to play a crucial role in A/I training as it provides unique opportunities to evaluate serious life threatening diseases. An unchanged trend of consultation for immunodeficiency and anaphylaxis reaffirms the importance of the allergist/immunologist as a valuable resource for inpatient consultation. PMID- 12722962 TI - The effects of acute doses of fexofenadine, promethazine, and placebo on cognitive and psychomotor function in healthy Japanese volunteers. AB - BACKGROUND: Genetic variations in cross-cultural metabolic capability may attenuate the lack of central nervous system effects of fexofenadine. OBJECTIVE: To compare the pharmacodynamics of fexofenadine and promethazine versus placebo in Japanese volunteers. METHODS: In this randomized, crossover, double-blind study, 24 subjects received single doses of fexofenadine 60 mg and 120 mg, promethazine 25 mg, and placebo, with a 6-day washout period between treatments. Objective measures included critical flicker fusion, choice reaction time, and a compensatory tracking task. A line analog rating scale evaluated self-rated sedation. A rapid visual information-processing task evaluated vigilance at baseline and at 2 hours. RESULTS: Fexofenadine was not significantly different from placebo on any test at any timepoint. In contrast, promethazine impaired critical flicker fusion thresholds (F[3,63] = 5.37, P = 0.0023); increased recognition reaction time (F[3,63] = 13.63, P < 0.0001) and total reaction time (F[3,63] = 12.23, P < 0.0001) components of the choice reaction time test; reduced tracking accuracy (F[3,63] = 14.25, P < 0.0001) and increased reaction times to peripheral stimuli (F[3,63] = 9.29, P < 0.0001) in the compensatory tracking task; reduced the number of valid responses (F[3,63] = 14.86, P < 0.0001) and impaired reaction times (F[3,63] = 12.02, P < 0.0001) in the rapid visual information-processing task test; and impaired subjective ratings of sedation (F[3,63] = 7.55, P = 0.0002), compared with placebo. CONCLUSIONS: A battery of tests sensitive to impairment by promethazine failed to show any negative cognitive or psychomotor effects with fexofenadine 60 and 120 mg. Fexofenadine is an intrinsically non-impairing antihistamine in Japanese subjects. PMID- 12722963 TI - Direct clinician-to-patient feedback discussion of inhaled steroid use: its effect on adherence. AB - STUDY OBJECTIVES: To evaluate whether direct feedback discussion on inhaled steroid use might influence subsequent adherence with this therapy. DESIGN AND SETTING: A 10-week, single-blind, randomized trial in asthma patients. Inclusion criteria included forced expiratory volume in 1 second <80%, one or more markers for low socioeconomic status, and the use of inhaled steroids. Inhaled steroid and beta-agonist use were electronically monitored. All patients received standard asthma care. The treatment group received direct clinician-to-patient feedback discussion on their inhaled steroid and beta-agonist use on all subsequent visits, whereas this information was withheld during the study period in the control group. MEASURES: 1) Mean weekly inhaled steroid adherence [(number of actuations/prescribed number of actuations) x 100]; 2) number of days with overuse of inhaled steroids; 3) 24-hour and nighttime albuterol use; 4) included forced expiratory volume in 1 second; and 5) Asthma Quality of Life Questionnaire total score. RESULTS: Ten treatment and nine control patients completed the study. Mean weekly inhaled steroid adherence over the first week was not significantly different in the treatment and control groups: 61 +/- 9% versus 51 +/- 5%, respectively. However, by the second week, adherence increased to 81 +/- 7% in the treatment group, whereas it decreased to 47 +/- 7% in the control group (P = 0.003). Adherence remained above 70% in the treatment group for the entire trial, but continued to decrease in the control group. Overuse of inhaled steroids was low in both groups. There were no group differences in any of the asthma outcomes. CONCLUSIONS: Direct clinician-to-patient feedback discussion on inhaled steroid use using electronic printouts did improve adherence in the short term in asthma patients at high-risk for poor adherence. PMID- 12722964 TI - Relief of cough and nasal symptoms associated with allergic rhinitis by mometasone furoate nasal spray. AB - BACKGROUND: Cough commonly occurs as a symptom of seasonal allergic rhinitis (SAR), an inflammatory condition of the nasal mucous membranes that results in rhinorrhea, nasal stuffiness/congestion, nasal itching, and sneezing. Mometasone furoate nasal spray (MFNS, Nasonex, Schering, Kenilworth, NJ), an anti inflammatory nasal corticosteroid, has been shown to be safe and effective in reducing the nasal inflammation of SAR. OBJECTIVE: To examine the effectiveness of MFNS in relieving SAR-associated cough, in addition to nasal symptoms. METHODS: This was a multicenter, randomized, double-blind study. Patients 12 years of age or older with > or = 1-year history of SAR symptoms, positive skin test to a prevailing seasonal allergen, moderate nasal symptoms, and moderate cough were treated for 14 days with MFNS 200 microg daily (n = 122) or placebo (n = 123). RESULTS: The group treated with MFNS showed significant improvement in the daytime cough severity score at endpoint compared with placebo (P = 0.049). Improvement in the nighttime cough severity score showed a trend in favor of MFNS treatment. Treatment with MFNS significantly improved total nasal symptoms in both the daytime and nighttime compared with placebo at endpoint (P < or = 0.017). Overall daytime symptom scores (cough + total nasal) improved significantly compared with placebo at endpoint (P = 0.005). Overall nighttime symptom scores improved significantly compared with placebo at endpoint (P = 0.028). Treatments were well tolerated, with no significant differences in the incidence of adverse events. CONCLUSIONS: MFNS is effective and well tolerated in the treatment of daytime cough associated with SAR. PMID- 12722965 TI - Serum interferon-gamma is associated with longitudinal decline in lung function among asthmatic patients: the Normative Aging Study. AB - BACKGROUND: Cytokines are important mediators of the asthmatic response. A retrospective pilot study showed that serum levels of interleukin (IL)-5 and interferon (IFN)-gamma were related to lung function decline among asthmatic patients over the preceding 3 years. To confirm these findings, we tested the hypothesis that serum cytokines are associated with longitudinal lung function decline. METHODS: We conducted a prospective, longitudinal study of 25 asthmatic and 50 nonasthmatic men (median age, 63 years; range, 45 to 80 years) participating in the Normative Aging Study. All study subjects completed two consecutive triennial examinations, including spirometry, methacholine challenge testing, allergy skin testing, and phlebotomy. Serum levels were measured for IL 4, IL-5, IL-6, IL-8, IL-10, and IFN-gamma. RESULTS: Among asthmatic patients, a higher initial serum level of IFN-gamma was associated with a greater rate of decline of forced expiratory volume in 1 second (FEV1; beta = -67 mL/year per log increase in serum IFN-gamma, P = 0.04) and, to a lesser extent, of FEV1/forced vital capacity ratio (beta = -0.91%/year per log increase in serum IFN-gamma, P = 0.07) after adjusting for age, smoking status, and baseline level of lung function. Serum IL-5 level was associated with a rate of decline in FEV1 of borderline significance (beta = -61 mL/year per log increase in serum IL-5, P = 0.08) among asthmatic patients. These relationshipswere not observed among nonasthmatic patients. CONCLUSIONS: Serum levels of IFN-gamma are associated with subsequent rate of change in lung function among asthmatic patients in this cohort of middle-aged and older men, and may be useful as biologic markers of risk for accelerated lung function decline in population studies. PMID- 12722966 TI - Interleukin-4 and interleukin-4 soluble receptor alpha levels in bronchoalveolar lavage from children with asthma. AB - BACKGROUND: In asthma there is increased expression of the Th2-type cytokine interleukin-4 (IL-4). IL-4 is important in immunoglobulin isotype switching to immunoglobulin E and adhesion of eosinophils to endothelium. OBJECTIVE: We hypothesized that levels of IL-4 in bronchoalveolar lavage (BAL) fluid would be increased in stable, atopic asthmatic children compared with controls and that levels of its physiologic inhibitor IL-4 soluble receptor alpha (IL-4sR alpha) would be correspondingly decreased. METHODS: One hundred sixteen children attending a children's hospital for elective surgery were recruited. A nonbronchoscopic BAL was performed, and IL-4 and IL-4sR alpha were measured in the BAL supernatants. RESULTS: There was no significant difference in IL-4 concentrations between atopic asthmatic children, atopic normal controls, and nonatopic normal controls [0.13 pg/mL (0.13 to 0.87) vs 0.13 pg/mL (0.13 to 0.41) vs 0.13 pg/mL (0.13 to 0.5), P = 0.65]. IL-4sR alpha levels were significantly increased in asthmatic patients compared with atopic controls [6.4 pg/mL (5.0 to 25.5) vs 5.0 pg/mL (5.0 to 9.9), P = 0.018], but not when compared with the nonatopic controls [5.2 pg/mL (5.0 to 10.6), P = 0.19]. CONCLUSIONS: Contrary to expectation, IL-4sR alpha levels are increased in BAL from stable asthmatic children compared with nonatopic controls, and we speculate that IL-4sR alpha is released by inflammatory cells in the airways to limit the proinflammatory effects of IL-4. PMID- 12722967 TI - Rapid anti-inflammatory action of azelastine eyedrops for ongoing allergic reactions. AB - BACKGROUND: Some second-generation antihistamines have anti-inflammatory activities, but the clinical relevance of this property is still unclear. OBJECTIVE: The aim of our study was to investigate the effects of azelastine when administered during the early-phase reaction. METHODS: This investigation was designed as a randomized, placebo-controlled, double-blind, parallel-group study. Clinical and inflammatory events were evaluated after a single dose of azelastine eyedrops or placebo was administered 30 minutes after an allergen-specific conjunctival challenge. Twenty outpatients with allergic rhinoconjunctivitis attributable to Parietaria judaica were enrolled in the study outside the pollen season. Patients were evaluated at baseline, after allergen challenge (at 30 minutes), and after administration of azelastine (at 30 minutes and at 6 hours). The following variables were evaluated: hyperemia, lacrimation, itching, eyelid swelling, number of inflammatory cells (neutrophils, eosinophils, monocytes, and lymphocytes), and intercellular adhesion molecule-1 expression on conjunctival epithelial cells. RESULTS: Azelastine, in comparison to placebo, significantly reduced symptom scores, number of inflammatory cells, and intercellular adhesion molecule-1 expression during the early- and late-phase reaction. CONCLUSIONS: The ability of azelastine to reduce symptoms and inflammation during an ongoing allergic reaction can be considered concrete and convincing proof of a clinically relevant anti-inflammatory activity. PMID- 12722969 TI - Anaphylactic reaction after artificial insemination. AB - BACKGROUND: Bovine seroalbumin is known as an allergen for human beings, but reactions to it in an artificial insemination procedure are much rarer. We report a case of anaphylaxis after intrauterine insemination (IUI) in which sensitization to bovine serum albumin (BSA) is demonstrated. OBJECTIVE: Report the allergy evaluation performed in a patient who suffered a severe reaction immediately after an IUI procedure. METHODS: A 33-year-old woman was referred because of an anaphylactic reaction after a second trial of IUI. She developed pruritus, abdominal pain, nausea and vomiting, bronchospasm, and generalized urticaria. She had an atopic medical history of pollen allergy and sensitization to cat epithelium. She had never had trouble with minor surgery and she usually uses latex material. She had never received heterologous sera before. Her husband's semen for the IUI was processed in a standard fluid medium called upgraded INRA B 2 (Laboratoires CCD, Paris, France), which contains amino acids, lipids, vitamins, BSA, penicillin, and streptomycin in addition to inorganic salts. RESULTS: Skin prick tests with the medium and BSA 10 mg/mL were positive. In vitro studies demonstrated an immunoglobulin E binding protein of 60 to 65 kDa and mast cells and basophil activation (CD63 expression) against BSA contained in the medium. Cutaneous and challenge tests with penicillin and streptomycin were negative. CONCLUSIONS: We consider the BSA in the semen culture medium to be the factor which triggered the anaphylactic reaction. This case supports the authors who state that media free from heterologous proteins should be used for human application, especially on atopic patients, to avoid sensitization. PMID- 12722970 TI - Cytokine abnormalities in a patient with eosinophilic fasciitis. AB - BACKGROUND: Eosinophilic fasciitis (EF) and peripheral eosinophilia are rare in children. OBJECTIVE: To describe a case of EF in a 3-year-old child who presented with acute painless induration of her forearm. METHODS: Cytokine profiles were obtained and compared with patients with atopic disease during the acute presentation and after treatment with low-dose prednisone. RESULTS: The patient's serum showed elevation of transforming growth factor beta1 and interleukin-5, which improved after treatment with low-dose prednisone. CONCLUSIONS: The case suggests that patients with EF have cytokine abnormalities similar to atopic patients, but with a striking elevation of transforming growth factor beta1. The responsiveness of the clinical symptoms and cytokine profile to low dose prednisone supports treatment with low dose immunosuppressive therapy in this disorder. PMID- 12722968 TI - Lipid transfer protein from Hevea brasiliensis (Hev b 12), a cross-reactive latex protein. AB - BACKGROUND: Latex-allergic individuals experience clinical cross-reactivity to a large number of fruits and vegetables. Much of the cross-reactivity can be attributed to Hev b 6, but evidence indicates that additional cross-reactive allergens may be present. A common pan-allergen, which has not previously been identified in latex, but may contribute to this cross-reactivity is lipid transfer protein (LTP). We sought to determine whether Hevea brasiliensis produces LTP and whether it would bind immunoglobulin E from latex-allergic patients. METHODS: LTP was identified in H. brasiliensis RNA by polymerase chain reaction using degenerate primers. The entire cDNA was obtained by polymerase chain reaction using rapid amplification of cDNA ends reactions. The complete coding sequence for LTP was determined and produced as a recombinant protein using the glutathione S-transferase and pET32 expression systems. Immunoblot analysis of sera from latex-allergic patients was used to determine whether patients recognize LTP as an allergen. RESULTS: We identified a 662-basepair cDNA with a 351-basepair open reading frame that encodes for a 116-amino acid protein. The protein has significant homology to the family of nonspecific LTPs. We expressed the protein as a mature LTP of 92 amino acids with a predicted isoelectric point of 10.8 and molecular weight of 9.3 kDa. Immunoblots demonstrated specific immunoglobulin E for LTP in the sera of 9 of 37 (24%) latex allergic individuals. CONCLUSIONS: We describe the initial identification of rLTP in H. brasiliensis that may be important as a cross-reactive pan-allergen (Hev b 12). PMID- 12722971 TI - Alcohol-induced memory impairment in trace fear conditioning: a hippocampus specific effect. AB - It has been hypothesized that the amnesic effects of alcohol are through selective disruption of hippocampal function. Delay and trace fear conditioning are useful paradigms to investigate hippocampal-dependent and independent forms of memory. With delay fear conditioning, learning of explicit cues does not depend on normal hippocampal function, whereas learning explicit cues in trace fear conditioning does. In both delay and trace fear conditioning, the hippocampus is involved in learning to contextual cues, but it may not be entirely necessary. The present study investigates the effects of alcohol on the acquisition of delay and trace fear conditioning in mice, using freezing as a measure of learning. Male C57BL/6J mice were injected with 0.8 or 1.6 g/kg of 20% v/v alcohol and were immediately exposed to eight tone-footshock pairings in which the conditional stimulus (CS) either coterminated with a footshock unconditional stimulus (US) (delay conditioning) or was separated from the footshock by a 30-s trace interval (trace conditioning). During trace, but not delay fear conditioning, 0.8 g/kg alcohol impaired learning to a tone CS. This dose also impaired context-dependent learning in both procedures (although only slightly for trace fear conditioning). The 1.6 g/kg alcohol exerted a nonselective impairment on learning. The impairment by alcohol of learning to a tone CS when it is hippocampus-dependent, but not when it is hippocampus independent provides further support for the hypothesis that alcohol exerts a selective effect on hippocampus-dependent learning. PMID- 12722972 TI - Function of hippocampus in "insight" of problem solving. AB - Since the work of Wolfgang Kohler, the process of "insight" in problem solving has been the subject of considerable investigation. Yet, the neural correlates of "insight" remain unknown. Theoretically, "insight" means the reorientation of one's thinking, including breaking of the unwarranted "fixation" and forming of novel, task-related associations among the old nodes of concepts or cognitive skills. Processes closely related to these aspects have been implicated in the hippocampus. In this research, the neural correlates of "insight" were investigated using Japanese riddles, by imaging the answer presentation and comprehension events, just after participants failed to resolve them. The results of event-related functional magnetic resonance imaging (fMRI) analysis demonstrated that the right hippocampus was critically highlighted and that a wide cerebral cortex was also involved in this "insight" event. To the best of our knowledge, this work is the first neuroimaging study to have investigated the neural correlates of "insight" in problem solving. PMID- 12722974 TI - Transitivity, flexibility, conjunctive representations, and the hippocampus. I. An empirical analysis. AB - After training on a set of four ordered, simultaneous, odor discrimination problems (A+B-, B+C-, C+D-, D+E), intact rats display transitivity: When tested on the novel combination BD, they choose B. Rats with damage to the hippocampus, however, do not show transitivity (Dusek and Eichenbaum, 1997. Proc Natl Acad Sci U S A 94:7109-7114). These results have been interpreted as support for the idea that the hippocampus is a relational memory storage system that enables the subject to make comparisons among representations of the individual problems and choose based on inferential logic. We provide evidence for a simpler explanation. Specifically, subjects make their choices based on the absolute excitatory value of the individual stimuli. This value determines the ability of that stimulus to attract a response. This conclusion emerged because after training on a five problem set (A+B-, B+C-, C+D-, D+E-, E+F-) rats preferred B when tested with BE, but not when tested with BD. The implication of these results for how to conceptualize the role of the hippocampus in transitive-like phenomena is discussed. PMID- 12722975 TI - Transitivity, flexibility, conjunctive representations, and the hippocampus. II. A computational analysis. AB - A computational neural network model is presented that explains how the hippocampus can contribute to transitive inference performance observed in rats (Dusek and Eichenbaum, 1997. Proc Natl Acad Sci U S A 94:7109-7114; Van Elzakker et al., 2003. Hippocampus 12:this issue). In contrast to existing theories that emphasize the idea that the hippocampus contributes by flexibly relating previously encoded memories, we find that the hippocampus contributes by altering the elemental associative weights of individual stimulus elements during learning. We use this model to account for a range of existing data and to make a number of distinctive predictions that clearly contrast these two views. PMID- 12722973 TI - Involuntary, unreinforced (pure) spatial learning is impaired by fimbria-fornix but not by dorsal hippocampus lesions. AB - Pure spatial learning occurs when rats acquire information about an environment while exploring it in the absence of reinforcers. We previously reported that voluntary, unreinforced exploration of a radial maze retards subsequent reinforced conditioned cue preference (CCP) learning in the same maze. In the present experiment, we examined the effects of involuntary, unreinforced pre exposure to a radial maze. During pre-exposure, rats were moved by an experimenter between the ends of two arms of a radial maze five times in 30 min. This form of pre-exposure retarded CCP learning, whereas rats that were not pre exposed and rats that were pre-exposed to a maze in a different room displayed normal CCP learning. These findings suggest that some information specific to the maze environment was acquired during involuntary unreinforced pre-exposure to it. In experiment 2, the retardation of reinforced CCP learning by involuntary unreinforced pre-exposure was eliminated by fimbria-fornix lesions made before pre-exposure but was unaffected by fimbria-fornix lesions made after pre-exposure but before training. Large neurotoxic lesions of the dorsal hippocampus made before pre-exposure had no effect on the retardation of CCP learning, but the rats with these lesions were impaired on a standard test of reinforced spatial learning in a water maze. The lesion effects in experiment 2 are similar to those previously reported for voluntary exploration and suggest that pure spatial learning may occur during both voluntary exploration of and involuntary exposure to an environment in the absence of reinforcers. Pure spatial learning can apparently occur with exposure to two different locations within an environment, but the rats do not have to move between the locations voluntarily. An intact fimbria-fornix is required for acquisition but not expression of this form of learning. The hippocampus is not involved in this form of learning. PMID- 12722976 TI - Phorbol ester uncouples adenosine inhibition of presynaptic Ca2+ transients at hippocampal synapses. AB - Synaptic transmission involves Ca2+ influx at presynaptic terminals. Adenosine receptors inhibit transmission, and this effect can be abolished by activation of PKC with phorbol esters. Whether protein kinase C (PKC) acts via alterations in Ca2+ entry at the presynaptic terminal is unknown. In the present study, we recorded the presynaptic Ca2+ transients (preCa(delta)) in hippocampal stratum radiatum, using fluorescence photometry. The calcium dye Fura-2 AM was used to load the Schaffer collateral/commissural tract and its terminals. Tetrodotoxin (TTX)-sensitive Na+ channels and Cd2+-sensitive, high-voltage activated Ca2+ channels (HVACCs) were required to elicit the preCa(delta). Application of the phorbol ester phorbol-12,13-dibutyrate (PDBu) abolished the adenosine inhibition of both preCa(delta) and the field excitatory postsynaptic potentials (fEPSPs). PDBu consistently potentiated fEPSPs, and also increased preCa(delta) in a large majority of the slices examined. Regardless of whether potentiation was observed, PDBu always prevented adenosine inhibition of preCa(delta). In contrast, the inactive phorbol ester, 4alpha-phorbol, did not alter adenosine inhibition of preCa(delta), indicating that PKC activation is necessary for the occurrence of the observed effects. Our findings suggest that PKC activation abolishes adenosine's inhibitory effect on synaptic activity involving presynaptic Ca2+ entry. PMID- 12722977 TI - Multiple hypothalamic sites control the frequency of hippocampal theta rhythm. AB - Stimulation of a neural pathway originating in the brainstem reticular formation, with synapses in the medial hypothalamus, activates the hippocampal theta rhythm. The frequency of reticular-elicited theta is determined in the medial supramammillary nucleus (mSuM) completely in anaesthetised rats, but only partially when the animal is awake. We tested other medial hypothalamic sites for their capacity to control theta frequency in awake rats. Blockade of sodium channels (1 microl fast infusion of the local anaesthetic procaine, experiment 1) or increased inhibition by GABA (Chlordiazepoxide [CDP], experiment 2) was found to reduce or increase the frequency of reticular-elicited theta, depending on the precise site of injection, in the region of the dorsomedial hypothalamic nucleus (DMH) and the posterior hypothalamic nucleus (PH). A band of null sites for CDP was located in the region of the ventral border of PH and dorsal border of mSuM. Using 0.5 and 1 microl CDP, and slow infusions (experiment 3), it was found that effective PH sites were also separate from mSuM in the rostrocaudal direction. In experiment 4, the DMH/PH region was mapped with unilateral and bilateral slow infusions of 0.5 microl CDP. CDP significantly reduced frequency in medial (periventricular) and dorsal PH, but not DMH. Bilateral injections appeared to generally sum the usual effects of unilateral injection, producing effects of intermediate size. However, the absolute frequency change in any given site, or with any pair of sites, did not exceed 1 Hz, which is similar to what is seen with single injections in mSuM. Overall, it appears that at, any one time, theta frequency may be determined by a complex interplay between distinct but interacting modulatory regions in the medial hypothalamus. PMID- 12722978 TI - Protective role of melatonin in domoic acid-induced neuronal damage in the hippocampus of adult rats. AB - Domoic acid (DA), a kainite-receptor agonist and potent inducer of neurotoxicity, has been administered intravenously in adult rats in the present study (0.75 mg/kg body weight) to demonstrate neuronal degeneration followed by glial activation and their involvement with inducible nitric oxide synthase (iNOS) in the hippocampus. An equal volume of normal saline was administered in control rats. The pineal hormone melatonin, which protects the neurons efficiently against excitotoxicity mediated by sensitive glutamate receptor, was administered intraperitoneally (10 mg/kg body weight), 20 min before, immediately after, and 1 h and 2 h after the DA administration, to demonstrate its role in therapeutic strategy. Histopathological analysis (Nissl staining) demonstrated extensive neuronal damage in the pyramidal neurons of CA1, CA3 subfields and hilus of the dentate gyrus (DG) in the hippocampus at 5 days after DA administration. Sparsely distributed glial fibrillary acidic protein (GFAP)-immunoreactive astrocytes were observed in the hippocampus at 4-24 h after DA administration and in the control rats. Astrogliosis was evidenced by increased GFAP immunoreactivity in the areas of severe neuronal degeneration at 5 days after DA administration. Along with this, microglial cells exhibited an intense immunoreaction with OX-42, indicating upregulation of complement type 3 receptors (CR3). Ultrastructural study revealed swollen or shrunken degenerating neurons in the CA1, CA3 subfields and hilus of the DG and hypertrophied astrocytes showing accumulation of intermediate filament bundles in the cytoplasm were observed after administration of DA. Although no significant change could be observed in the mRNA level of iNOS expression between the DA-treated rats and controls at 4-24 h and at 5-day time intervals, double immunofluorescense revealed co-expression of induced iNOS with GFAP immunoreactive astrocytes, but not in the microglial cells, and iNOS expression in the neurons of the hippocampal subfields at 5 days after DA administration. Expression of iNOS was not observed in the hippocampus of control rats. DA induced neuronal death, glial activation, and iNOS protein expression were attenuated significantly by melatonin treatment and were comparable to the control groups. The results of the present study suggest that melatonin holds potential for the treatment of pathologies associated with DA-induced brain damage. It is speculated that astrogliosis and induction of iNOS protein expression in the neurons and astrocytes of the hippocampus may be in response to DA-induced neuronal degeneration. PMID- 12722979 TI - Effects of alcohol exposure during early life on neuron numbers in the rat hippocampus. I. Hilus neurons and granule cells. AB - We previously showed that 16-day-old rats exposed to a relatively high dose of ethanol at 10-15 postnatal days of age have fewer neurons in the hilus region of the hippocampus compared with controls. Dentate gyrus granule cell numbers, however, showed no statistically significant changes attributable to the ethanol treatment. It is possible that some of the changes in brain morphology, brought about as a result of the exposure to ethanol during early life, may not be manifested until later in life. This question has been further addressed in an extension to our previous study. Wistar rats were exposed to a relatively high daily dose of ethanol on postnatal days 10-15 by placement in a chamber containing ethanol vapour, for 3 h/day. The blood ethanol concentration was found to be approximately 430 mg/dl at the end of the period of exposure. Groups of ethanol-treated (ET), separation control (SC), and mother-reared control (MRC) rats were anaesthetised and killed either at 16 or 30 days of age by perfusion with phosphate-buffered 2.5% glutaraldehyde. The Cavalieri principle and the physical disector methods were used to estimate, respectively, the regional volumes and neuron cell numerical densities in the hilus and granule cell regions of the dentate gyrus. The total numbers of neurons in the hilus region and granule cell layer were computed from these estimates. It was found that 16-day old animals had 398,000-441,000 granule cells, irrespective of group. The numbers of granule cells increased such that by 30 days of age, rats had 487,000-525,500 granule cells. However, there were no significant differences between ethanol treated rats and their age-matched controls in granule cell numbers. In contrast, ethanol-treated rats had slightly but significantly fewer neurons in the hilus region than did control animals at 16 days of age, but not at 30 days of age. Therefore, it appears that a short period of ethanol exposure during early life can have effects on neuron numbers of some hippocampal neurons, but not others. The effects on hilar neuron numbers, observed as a result of such short periods of ethanol treatment, appeared to be transitory. PMID- 12722981 TI - Maturation of granule cell dendrites after mossy fiber arrival in hippocampal field CA3. AB - Most granule neurons in the rat dentate gyrus are born over the course of the first 2 postnatal weeks. The resulting heterogeneity has made it difficult to define the relationship between dendritic and axonal maturation and to delineate a time course for the morphological development of the oldest granule neurons. By depositing crystals of the fluorescent label Dil in hippocampal field CA3, we retrogradely labeled granule neurons in fixed tissue slices from rats aged 2-9 days. The results showed that all labeled granule cells, regardless of the age of the animal, exhibited apical dendrites. On day 2, every labeled neuron had rudimentary apical dendrites, and a few dendrites on each cell displayed immature features such as growth cones, varicosities, and filopodia. Some cells displayed basal dendrites. By day 4, the most mature granule neurons had longer and more numerous apical branches, as well as various immature features. Most had basal dendrites. On days 5 and 6, the immature features and the basal dendrites had begun to regress on the oldest cells, and varying numbers of spines were present. On day 7, the first few adult-like neurons were seen: immature features and basal dendrites had disappeared, all dendrites reached the top of the molecular layer, and the entire dendritic tree was covered with spines. These data show that dendritic outgrowth occurs before, or concurrent with, axon arrival in the CA3 target region, and that adult-like granule neurons are present by the end of the first week. PMID- 12722982 TI - The ketogenic diet for the treatment of epilepsy: a challenge for nutritional neuroscientists. AB - The ketogenic diet (KD) is a high-fat, low-carbohydrate, adequate-protein diet that has been used for more than eight decades for the treatment of refractory epilepsy in children. Despite this long history, the mechanisms by which the KD exerts its anti-seizure action are not fully understood. Questions remain regarding several aspects of KD action, including its effects on brain biochemistry and energetics, neuronal membrane function and cellular network behavior. With the explosion of the KD use in the last 10 years, it is now imperative that we understand these factors in greater detail, in order to optimize the formulation, administration and fine-tuning of the diet. This review discusses what is known and what remains to be learned about the KD, with emphasis on clinical questions that can be approached in the laboratory. We encourage scientists with a primary interest in nutritional neuroscience to join with those of us in the epilepsy research community to address these urgent questions, for the benefit of children ravaged by intractable seizures. PMID- 12722983 TI - Gastric emptying and control of ingestion in preweanling rat pups. AB - In adult rats, the rate of gastric emptying is modulated by properties of the diet, including its caloric content and osmotic properties. In developing pups, there is little known about the modulation of gastric emptying, despite evidence that volume of gastric contents may play a significant role in modulating intake in young rats. The present experiments examined gastric emptying of oral infusions of corn oil emulsions or glucose solutions in pups aged 6 or 15 days of age and their effects on independent ingestion. The results demonstrated that pups as young as 6 days of age modulate the rate of gastric emptying in response to changes in the concentration of a corn oil or glucose diet. However, oral infusions of corn oil emulsions failed to produce differences in subsequent intake of a milk diet, while oral infusions of a glucose diet did suppress subsequent intake. Taken together, the results support different ontogenetic courses for the development of mechanisms modulating gastric emptying of lipid versus carbohydrate diets, and suggest that gastric fill plays a limited role in modulation of intake in young pups. PMID- 12722984 TI - The effects of phenylpropanolamine on Zucker rats selected for fat food preference. AB - Treatments of human and rodent obesity frequently involve administration of amphetamine derivatives, much like phenylpropanolamine, which suppress food intake. The Zucker rat is a commonly employed model of youth-onset obesity in which the homozygous genotype manifests hyperphagia as well as other characteristics that parallel human obesity. Using a macronutrient selection procedure, we examined phenylpropanolamine's differential actions in controlling dietary intake, spontaneous open-field activity, and regional hypothalamic neurotransmitter levels in obese female Zucker rats of varying fat food preference. We hypothesized that phenylpropanolamine would alter hypothalamic monoamine levels differently in low-fat preferring and high-fat preferring Zucker rats, and hence affect feeding behavior and activity differently in these two groups. It was found that in high-fat preferring animals, phenylpropanolamine significantly decreased spontaneous open-field activity, decreased only carbohydrate caloric intake, and increased serotonin and 5-HIAA levels in the paraventricular nucleus (PVN). In low-fat preferring animals, phenylpropanolamine decreased carbohydrate, protein, and total caloric intake, had no significant effect of spontaneous activity, and increased serotonin and 5-hydroxyindole acetic acid levels in the PVN. Inherent and induced physiological differences of low-fat and high-fat preferring animals are discussed as well as phenylpropanolamine's potential in combination drug therapy for the treatment of human hyperphagic obesity. PMID- 12722985 TI - Sibutramine decreases body weight gain and increases energy expenditure in obese Zucker rats without changes in NPY and orexins. AB - The aim of the present work was to describe the effects of sibutramine on body weight and adiposity and to establish the potential involvement of neuropeptide Y (NPY) and orexins in the anorectic action of this drug. Male obese Zucker rats were daily administered with sibutramine (10 mg/kg, intraperitoneal) for two weeks. Carcass composition was assessed using the official methods of the Association of Official Analytical Chemists. Total body oxygen consumption was measured daily for 60 min before sibutramine or saline injection and for 30 min (from 60 to 90 min) after drug or saline injection. Hypothalamic arcuate and paraventricular nuclei, and the lateral hypothalamic area were immunostained for NPY, orexin A and orexin B. Commercial kits were used for serum determinations. Reductions in body weight and adipose tissue weights were observed after sibutramine treatment in obese Zucker rats. No changes in NPY immunostaining in the arcuate and paraventricular nuclei were found. Orexin A and orexin B immunostaining was not modified in the lateral hypothalamic area in treated rats. The reduction in body weight and adiposity induced by sibutramine was achieved by both a reduction in food intake and an increase in energy expenditure. NPY and orexins do not seem to be involved in the anorectic effect of sibutramine. PMID- 12722986 TI - A paradigm of undernourishing and neonatal rehabilitation in the newborn rat. AB - Perinatal undernutrition as a deficiency of nutrient availability, affects body and brain developmental processes and promotes recurrent health problems. Thus, altered mother-litter bonds and deficient environmental interactions may interfere with the brain pluripotential capabilities of the newborn. To gather information concerning the mechanisms underlying perinatal undernutrition we designed a paradigm of undernutrition and neonatal rehabilitation in the rat. An underfed group came from pregnant Wistar rats fed with 50% of the diet from G6 to G12 and with 60% from G13 until G21. After birth, pups were daily undernourished during 12 h daily by rotating a pair of lactating well-nourished dams which had one of their nipples subcutaneously ligated. The rehabilitated animals were undernourished pups neonatally fed by a pair of normally lactating dams. Controls received plenty of food during the pre- and neonatal periods. Pups were sacrificed at 12, 20 and 30 days of age. Perinatal underfeeding significantly reduced body and brain weights and neuronal morphometric parameters. Normal neonatal feeding in the newborn ameliorated the damages associated to food deprivation. The current undernourishing paradigm may be helpful to assess brain development alterations, as well as to study the compensatory mechanisms associated to salutary epigenetic influences. PMID- 12722980 TI - Mossy fiber plasticity and enhanced hippocampal excitability, without hippocampal cell loss or altered neurogenesis, in an animal model of prolonged febrile seizures. AB - Seizures induced by fever (febrile seizures) are the most frequent seizures affecting infants and children; however, their impact on the developing hippocampal formation is not completely understood. Such understanding is highly important because of the potential relationship of prolonged febrile seizures to temporal lobe epilepsy. Using an immature rat model, we have previously demonstrated that prolonged experimental febrile seizures render the hippocampus hyperexcitable throughout life. Here we examined whether (1) neuronal loss, (2) altered neurogenesis, or (3) mossy fiber sprouting, all implicated in epileptogenesis in both animal models and humans, were involved in the generation of a pro-epileptic, hyperexcitable hippocampus by these seizures. The results demonstrated that prolonged experimental febrile seizures did not result in appreciable loss of any vulnerable hippocampal cell population, though causing strikingly enhanced sensitivity to hippocampal excitants later in life. In addition, experimental febrile seizures on postnatal day 10 did not enhance proliferation of granule cells, whereas seizures generated by kainic acid during the same developmental age increased neurogenesis in the immature hippocampus. However, prolonged febrile seizures resulted in long-term axonal reorganization in the immature hippocampal formation: Mossy fiber densities in granule cell- and molecular layers were significantly increased by 3 months (but not 10 days) after the seizures. Thus, the data indicate that prolonged febrile seizures influence connectivity of the immature hippocampus long-term, and this process requires neither significant neuronal loss nor altered neurogenesis. In addition, the temporal course of the augmented mossy fiber invasion of the granule cell and molecular layers suggests that it is a consequence, rather than the cause, of the hyperexcitable hippocampal network resulting from these seizures. PMID- 12722987 TI - Extracellular hypothalamic serotonin and plasma amino acids in response to sequential carbohydrate and protein meals. AB - In previous studies, we showed that carbohydrate and protein ingestion, respectively, increased and decreased hypothalamic extracellular serotonin and the plasma ratio tryptophan over its competitor amino acids (Trp/LNAAs), reflecting serotonin synthesis. Serotonin levels returned towards baseline 2 h after either meal while the ratio remained altered. The question addressed is the ability of serotonin to respond expectedly to a second meal of the alternate nutrient. Rats were fed with sequential meals of either carbohydrates first and then casein 2 h later or in reverse order. Hypothalamic serotonin was measured using microdialysis. Permanent blood sampling allowed to track in parallel plasma amino acids. A carbohydrate meal increased hypothalamic serotonin, so did a subsequent casein meal. Conversely, following a casein meal that reduced serotonin, a carbohydrate meal also decreased it. The plasma ratio Trp/LNAAs was enhanced by a carbohydrate meal and remained high for 2h. A subsequent casein meal reversed this change but the ratio remained higher than basal values. A first casein meal reduced the ratio that was not increased again by a subsequent carbohydrate meal. It is obvious that ingestion of specific nutrients induce long lasting metabolic and neurochemical variations that prevent subsequent changes to occur. The lack of expected changes to a second meal addresses again the hypothesis of alternate appetites for carbohydrates and proteins driven by serotonin changes. PMID- 12722988 TI - Prolonged treatment with L-lysine and L-arginine reduces stress-induced anxiety in an elevated plus maze. AB - The aim of this study was to investigate whether a chronic pretreatment with a combination of L-lysine (Lys) and L-arginine (Arg) reduces anxiogenic effects of acute stress in rats. Male rats were orally infused with a distilled water solution of L-glutamine (200 mg/kg), Lys (200 mg/kg), or a combination of Lys (200 mg/kg) plus Arg (200 mg/kg) for four consecutive days (twice daily) and subjected to restraint stress on the fifth day. Immediately thereafter, rats were placed on an elevated plus maze (EPM) and their behavior was evaluated for 10 min. Lys and Arg significantly increased exploration time rats spent on open arms of the EPM, as compared to L-glutamine controls. In addition, the combination of Lys and Arg partly, but significantly, decreased stress-enhanced plasma corticosterone measured at the end of behavioral testing. Data suggest that a treatment with a solution of Lys and Arg reduces anxiety in stressed rats. PMID- 12722989 TI - Choline availability during embryonic development alters the localization of calretinin in developing and aging mouse hippocampus. AB - Choline availability in the diet during pregnancy alters fetal brain biochemistry with resulting behavioral changes that persist throughout the lifetime of the offspring. In the present study, the effects of dietary choline on the onset of GABAergic neuronal differentiation in developing fetal brain, as demarcated by the expression of calcium binding protein calretinin, are described. In these studies, timed-pregnant mice were fed choline supplemented, control or choline deficient AIN-76 diet from day 12-17 of pregnancy and the brains of their fetuses were studied on day 17 of gestation. In the primordial dentate gyrus, we found that pups from choline deficient-dams had more calretinin protein (330% increase), and pups from choline supplemented-dams had less calretinin protein (70% decrease), than did pups from control-dams. Importantly, decreased calretinin protein was still detectable in hippocampus in aged, 24-month-old mice, born of choline supplemented-dams and maintained since birth on a control diet. Thus, alterations in the level of calretinin protein in fetal brain hippocampus could underlie the known, life long effects of maternal dietary choline availability on brain development and behavior. PMID- 12722990 TI - Analyzing data in which the outcome is time to an event part II: The presence of multiple covariates. AB - In this article, the second of a series on the analysis of time to event data, we address the case in which multiple predictors (covariates) that may influence the time to an event are taken into account. The hazard function is introduced, and is given in a form useful for assessing the impact of multiple covariates on time to an event. Methods for the assessment of model fitting are also discussed and an example with cancer survival as outcome with the presence or absence of multiple genes as covariates is presented. PMID- 12722991 TI - Triumphs and controversies in laparoscopy: the past, the present, and the future. PMID- 12722992 TI - Telerobotic laparoscopic repair of incisional ventral hernias using intraperitoneal prosthetic mesh. AB - Laparoscopic ventral hernia repair shortens the length of hospital stay and achieves low rates of hernia recurrence. The inherent difficulties of performing advanced laparoscopy operations, however, have limited the adoption of this technique by many surgeons. We hypothesized that the virtual operative field and hand-like instruments of a telerobotic surgical system could overcome these limitations. We present herein the first 2 reported cases of telerobotic laparoscopic ventral hernia repair with mesh. The operations were accomplished with the da Vinci telerobotic surgical system. The hernia defects were repaired with dual-sided, expanded polytetrafluoroethylene (ePTFE) mesh. The mesh was secured in place with 8 sutures that were passed through the abdominal wall, and 5-mm surgical tacks were placed around the circumference of the mesh. The 2 operations were accomplished with total operative times of 120 and 135 minutes and total operating room times of 166 and 180 minutes, respectively. The patients were discharged home on postoperative days 1 and 4. The surgeon sat in an ergonomically comfortable position at a computer console that was remote from the patient. Immersion of the surgeon within the 3-dimensional virtual operative field expedited each stage of these procedures. The articulation of the wristed telerobotic instruments greatly facilitated reaching the anterior abdominal cavity near the abdominal wall. This report indicates that telerobotic laparoscopic ventral hernia repair is feasible and suggests that telepresence technology facilitates this procedure. PMID- 12722994 TI - Laparoscopic bowel resection: a comparison of three ultrasonically activated devices. AB - BACKGROUND AND OBJECTIVES: To compare resection time and collateral thermal damage of 3 currently available ultrasonically activated devices in laparoscopic small bowel surgery. METHODS: AutoSonix, SonoSurg, and UltraCision were compared in laparoscopic small bowel mesentery resection in a porcine model. A resection was defined as 12 endarcade arteries supplying the intended bowel segment. Vssels were divided 1 cm off the bowel wall. AutoSonix, SonoSurg, and UltraCision were comparable for blade length and type, cutting mechanism, handle ergonomics, and vibration amplitude, but not well matched for vibration frequency (55.5;23.5;55.5 kHz), working shaft diameter (5;11;10 mm) and length (29;33;34 cm), respectively. A sample size of 114 was calculated to detect a 25% difference with 90% power at a 5% significance level. Resections were allocated to devices by block randomization. Analysis of variance and pairwise Scheffe tests were used for multiple comparisons, and a Kaplan-Meier plot was drawn to confirm differences in resection time with each device. A pathologist blind to the devices evaluated bowel wall biopsies for thermal damage. RESULTS: Procedures as allocated comprised 114 resections (38 with each device). UltraCision median resection time of 5160 (range 2340-7860) seconds was significantly longer (P=0.0001). The difference in resection time between AutoSonix (median 3420, range 1860-8760 s) and SonoSurg (median 3660, range 1800-6900 s) did not reach statistical significance. A microscopy revealed no thermal damage. CONCLUSIONS: Laparoscopic resection time for porcine bowel mesentery was shorter with AutoSonix or SonoSurg than with UltraCision, and no thermal damage to the bowel wall was found. PMID- 12722995 TI - Recurrent cecocolic torsion: radiological diagnosis and treatment. AB - Recurrent cecocolic torsion may cause recurrent right lower abdominal pain and right-sided thrust dyspareunia. It is seldom considered in the differential diagnosis of pain in this region. The propensity of the cecocolon to torse is secondary to the double developmental defect of a mobile cecocolon compounded by an elongated and overrotated organ that can be eccentrically located in the abdomen. The torsion may result in recurrent obstructions with spontaneous resolutions, but it may proceed to an acute episode of obstruction and strangulation with a more profound morbidity and mortality rate. The diagnosis of recurrent cecocolic torsion is verified by a plain x-ray film of the abdomen, contrast enema, and computed tomography scan. The preferred treatment is outpatient laparoscopic cecocolopexy. Laparoscopic or classical open cecocolectomy and right hemicolectomy are reserved for more complex and morbid presentations. PMID- 12722993 TI - Accuracy of laparoscopic diagnosis of endometriosis. AB - BACKGROUND AND OBJECTIVE: Laparoscopy is the standard method to visually identify endometriotic lesions under magnification within and outside the minor pelvis. The aim of this study was to analyze the accuracy of laparoscopic visualization in diagnosing the various endometriotic sites as confirmed histologically. METHOD: Presumed endometriotic sites were observed in 164 patients operated on under the clinical suspicion of endometriosis. Targeted biopsies were performed for histologic corroboration, comparing the laparoscopic findings and diagnosis to the histological results. RESULT: The histological reports of the biopsies confirmed the presence of endometriosis in 138 patients (84.1%), but in 26 patients (15.9%), no evidence of endometriosis was observed. 100% of "red" lesions, 92% of "black" lesions, and 31% of "white" lesions turned out to be endometriosis. Of the 264 various suspected endometriotic sites observed, 142 (53.8%) were confirmed histologically. The most accurate diagnosis was in lesions on the parietal peritoneum of the pelvis, confirmed in 9/9 cases (100%); the ovarian fossa, confirmed in 8/12 cases (66.7%); and the uterosacral ligaments and posterior surface of the broad ligament, confirmed in 83/138 cases (60.1%). As for the other sites, the histologic confirmation rates in the ovarian surface, bowel serosa, and vesicouterine fold of the peritoneum were 48%, 40%, and 13%, respectively. CONCLUSION: Endometriosis has a multiple appearance, and the lesions may be confused with nonendometriotic lesions. It is clear that a nonhistology-based diagnosis may lead to unnecessary prolonged medical treatment and operations and may delay the proper treatment measures from being applied. Therefore, a meticulous histological confirmation should still be the first step in the laparoscopic diagnosis and treatment of suspected endometriosis. PMID- 12722996 TI - A modular laparoscopic training program for pediatric surgeons. AB - OBJECTIVES: A structured endoscopic training program for pediatric surgeons has not yet been established. This study was conducted to develop a modular training program (MTP) for pediatric surgeons and to evaluate its effectiveness for surgeons with and without previous experience in laparoscopic surgery. METHODS: Nine pediatric surgeons participated in the study. They were divided into 2 groups: group A (n=4), surgeons who had experienced more than 10 cases of laparoscopic surgery prior to MTP; group B (n=5), those who had experienced fewer than 10 cases. They participated in a standardized MTP workshop, which consisted of 2 "see-through" and 3 "laparoscopic" tasks. Each participant's psychomotor skills were evaluated objectively before and after MTP with a computer-generated virtual simulator and were evaluated for precision, efficiency, and speed. RESULTS: In participants, speed was significantly enhanced after MTP. In group A, no differences were observed after MTP, whereas significant improvements were noted in efficiency and speed after MTP in group B. Before MTP, efficiency was significantly higher in group A than in group B; however, no difference remained between the 2 groups after MTP. CONCLUSIONS: MTP is effective for nonlaparoscopic pediatric surgeons to become familiar with basic endoscopic skills. PMID- 12722997 TI - Hysteroscopic myomectomy: our experience and review. AB - OBJECTIVE: To analyze the results of hysteroscopic myomectomy in our center and to compare the results to those published in the literature. METHODS: We performed a retrospective study of the clinical histories of patients who had undergone hysteroscopic myomectomy with a resectoscope between January 1992 and December 1999. Procedures were performed at a hysteroscopic clinic in the Department of Obstetrics and Gynecology at the University Public Hospital in Madrid's south zone. One hundred twenty pre-, peri-, and postmenopausal women with submucous myomas were included in the study. All patients underwent hysteroscopic resection with a monopolar loop. RESULTS: We performed 120 hysteroscopic myomectomies. The patients' median age was 44.8 years (23 to 74). Abnormal uterine bleeding (AUB) was the most frequent indication (84.1%). Inability to reproduce was the indication in 14 (11.6%) cases. GnRH analogue preparation was used in 60% of cases. We operated on 52 (43.3%) type 0, 51 (42.5%) type I, and 17 (14.1%) type II myomas, according to Wamsteker and Blok classification. A median of 32.5 (10 to 105) minutes was required for the interventions. The myomectomy was combined with another operation (12 polypectomies, 24 endometrial resections, and 1 laparoscopic ovarian cystectomy) in 32 patients. The median retention of glycemia was 281 cc (0 to 1300). We could not complete the resection in 22 patients. Twelve underwent reoperation (3 hysterectomies and 9 second myomectomies). No serious complications occurred, and the median hospital stay was 25.4 hours. The histological study confirmed leiomyoma in all the cases. The intervention results were satisfactory after a follow-up period of 12 months to 7 years, AUB being controlled in 88.5% of the patients. CONCLUSION: Hysteroscopic myomectomy is a reliable procedure that is effective in controlling abnormal uterine bleeding. It is a good alternative to hysterectomy and has an acceptable surgical time and minimum hospital stay. To reduce the need of reintervention, appropriate patient selection and improved technique are necessary. The technique also offers significant economic savings compared with the conventional surgical methods. PMID- 12722998 TI - Introduction of a laparoscopic gynecological program in a general hospital in Taiwan. AB - BACKGROUND AND OBJECTIVES: We reviewed the records for 571 gynecologic laparoscopies performed at a privately owned general hospital in Kaosiung Taiwan in 1998 and 1999 and discuss here the major obstacles we encountered while introducing these procedures at our institution. METHODS: Included in this series are 293 procedures performed in 1998 (149 hysterectomies, 144 adnexal procedures), and 278 procedures performed in 1999 (131 hysterectomies, 147 adnexal procedures). Thirty-nine of these patients also underwent laparoscopic appendectomy. Mean patient age was 62 years (range 28 to 82). All procedures were performed by 1 of 6 board-certified gynecologic surgeons, or by 1 of 4 residents under the direct supervision of a board-certified surgeon. RESULTS: We experienced 0% mortality and 7.2% morbidity in this series. In comparing cases from 1998 and 1999, we observed a decrease in both mean surgery time (135.4 to 123.0, P=0.032) and mean length of hospital stay (5.52 to 4.62, P=0.046) for hysterectomies and adnexal procedures combined. CONCLUSIONS: These data support ongoing efforts to incorporate gynecologic laparoscopy as an alternative to open procedures at our institution. Introduction of these procedures in privately owned hospitals in Taiwan has been limited because of the large initial investment for equipment, patient education issues, and difficulties obtaining reimbursement. PMID- 12722999 TI - Complications of operative gynecological laparoscopy. AB - OBJECTIVE: To assess the incidence and type of laparoscopic complications. METHODS: A series of 2140 operative laparoscopies were reviewed in a retrospective study of patient records. The setting was a tertiary-care university hospital. Operative laparoscopy included minor procedures (minimal adhesiolysis, destruction of minimal endometriosis foci, ovarian biopsy, ovarian puncture, tubal sterilization), major laparoscopic surgery (extended adhesiolysis, tuboplasties, uterine suspension, treatment for ectopic pregnancy, salpingitis, ovarian cyst, moderate and severe endometriosis), and advanced laparoscopic surgery (hysterectomy, myomectomy, bladder neck suspension). RESULTS: Two major vascular complications, 3 intestinal injuries, 1 anesthesiological complication, and 4 urinary tract injuries occurred. Two minor and 5 postoperative complications were noted. The overall complication rate was 17/2140 (0.79%). The major complication rate was 10/2140 (0.46%). CONCLUSIONS: This review is useful for helping surgeons reduce the risk of injuries and to inform patients about potential complications. These rates are similar to those that have been previously reported. PMID- 12723000 TI - Robotically assisted endoscopic ovarian transposition. AB - BACKGROUND: Ovarian transposition is the anatomical relocation of the ovaries from the pelvis to the abdomen. Transposition is beneficial in women who are to undergo pelvic radiation, because it allows maintenance of ovarian function and preservation of assisted reproductive capacity. METHODS: The da Vinci surgical system (Intuitive Surgical, Mountainview, CA, USA) was used to perform an endoscopic ovarian transposition. The ovaries were mobilized on their respective infundibulopelvic ligaments and sutured to the ipsilateral pericolic gutters. RESULTS: A series, of laboratory sessions using the da Vinci system was completed at our institution's training facility. Surgical experience included cadaveric pelvic dissection and abdominopelvic procedures on anesthetized porcine models. Additional didactic and laboratory training, including a certification examination, was obtained from Intuitive Surgical, Inc. The first clinical case of robotically assisted endoscopic ovarian transposition was performed. CONCLUSIONS: Robotically assisted endoscopy was successfully used for ovarian transposition. PMID- 12723001 TI - Laparoscopic nephrectomy for renovascular hypertension in a 6-month-old infant. AB - Laparoscopic procedures continue to gain popularity over traditional open operations for a variety of abdominal and retroperitoneal surgical procedures. With regard to urological surgery, the first laparoscopic nephrectomy was performed in an adult in 1991. In the following years, the feasibility of laparoscopic management of pediatric urological disorders was described, and in 1992 the first laparoscopic nephrectomy in an 8-month-old infant with a multicystic dysplastic kidney was reported. We report the feasibility of laparoscopic nephrectomy for the management of renovascular hypertension in a 6 month-old infant with a dysplastic left kidney. PMID- 12723003 TI - Pneumomediastinum as a complication of preperitoneal laparoscopic herniorrhaphy. AB - BACKGROUND: As new approaches to herniorrhaphy are popularized, new complications are being encountered. Pneumomediastinum is one such complication. METHODS: We present a patient who developed pneumomediastinum after an elective bilateral preperitoneal laparoscopic hernia repair. RESULTS: The patient was observed and discharged 24 hours later with no permanent sequelae. CONCLUSION: Pneumomediastinum is a rare complication of laparoscopic hernia repair. PMID- 12723004 TI - Laparoscopic excision of a mesenteric cyst during pregnancy. AB - BACKGROUND: Laparoscopic management of mesenteric cysts during pregnancy has not been reported before. CASE REPORT: A young woman with a mesenteric cyst associated with a crossed ectopic kidney, underwent laparoscopic excision of the cyst in the second trimester of pregnancy. The procedure was completed without complications, and the patient was discharged on the third postoperative day. Thereafter, the pregnancy progressed uneventfully, and she delivered a healthy baby at term. CONCLUSION: Laparoscopic management of mesenteric cysts during pregnancy is feasible, safe, and less invasive than laparotomy when performed in select patients by experienced surgeons. PMID- 12723005 TI - Obesity and diabetes: is the worst yet to come for cardiovascular disease? PMID- 12723002 TI - Postoperative hypoxemia from clinically suspected pulmonary embolism complicated by patent foramen ovale. AB - Postoperative hypoxemia can be a challenging diagnostic and management dilemma for the clinician. We present here a case of postoperative hypoxemia following laparoscopic gastric bypass surgery secondary to presumed pulmonary embolism complicated with a patent foramen ovale. The diagnostic pitfalls associated with a negative spiral computed tomography scan and the impact of coexisting medical conditions aggravating the pulmonary dysfunction are reviewed. PMID- 12723006 TI - Cultural orientation and diabetes self-care in low-income African Americans with type 2 diabetes mellitus. AB - OBJECTIVES: The purpose of the study was to examine the relationships between cultural variables and diabetes self-care behaviors and glycemic control among African Americans with type 2 diabetes. DESIGN: Cross-sectional survey. METHODS: Questionnaires assessing traditional African-American cultural orientation, ethnic identity, self-identification, and diabetes self-care were administered to a sample of 94 low-income, African-American, inner-city hospital outpatients with type 2 diabetes. Participants were predominantly female (64%), with an average age of 53 years, and most had attained less than or equal to a high school education (66%). RESULTS: No significant relationships were found among ethnic identity, self-identification, glycemic control, and diabetes self-care behaviors. Traditional African-American cultural orientation was significantly associated with decreased dietary adherence scores (P<.03). Increased scores on cultural mistrust were related to decreased dietary adherence scores (P<.002). Traditional food practices showed a non-significant trend toward decreased dietary adherence in conjunction with number of dependents and income (P<.055). CONCLUSIONS: Traditional African-American cultural orientation was found to be associated with decreased dietary adherence scores in a sample of urban African Americans with type 2 diabetes. Assessment of the cultural orientation of African American patients has the potential to assist providers in designing culturally tailored, diabetes-specific dietary interventions. PMID- 12723008 TI - The effectiveness of a community health worker outreach program on healthcare utilization of west Baltimore City Medicaid patients with diabetes, with or without hypertension. AB - OBJECTIVES: Assess impact of community health workers (CHWs) on healthcare utilization of African-American Medicaid patients with diabetes mellitus (DM) with/without hypertension (HTN). DESIGN: Maryland Medicaid claim files were analyzed to compare utilization of emergency rooms, hospitalization, and costs in a population managed by CHWs (N=117). SETTING/PATIENTS: Patients were recruited from the discharge rolls of the University of Maryland Medical System (UMMS) and/or the Maryland Diabetes Care Program from March 1992 to June 1994. INTERVENTION: CHWs alternated weekly home visits and phone contacts to teach patients to understand the need to control their illnesses, to follow both their therapy and behavioral regimens, and to maintain appropriate visits to a primary care practitioner. RESULTS: Total emergency room (ER) visits declined by 40%; ER admissions to hospitals declined by 33%, as did total hospital admissions; and Medicaid reimbursements declined by 27%. CONCLUSIONS: The CHW program resulted in an average savings of 2,245 dollars per patient per year, and a total savings of 262,080 dollars for 117 patients, with improved quality of life (QOL) indicating cost effectiveness. PMID- 12723007 TI - A preliminary assessment of acculturation and its relationship to body size and glucose intolerance among Blacks in the US Virgin Islands. AB - OBJECTIVES: This study is a preliminary investigation of the relationship of acculturation to body size and glucose intolerance among African Caribbeans living in the United States Virgin Islands (USVI). METHODS: Focus groups were used to identify items for measuring acculturation, and validity and reliability procedures were performed on the items. The acculturation items were administered to a population-based sample of 740 individuals aged 20 years and older. Anthropometric measurements and blood pressure were taken and a fasting blood sample was also drawn. RESULTS: Factor analysis showed that the acculturation items clustered as 3 factors that appeared to represent: 1) the 'adoption' of USVI-American values; 2) 'integration' and practice of USVI customs; and 3) allegiance to 'traditional' African-Caribbean values and customs. Each factor exhibited relationships to demographic variables that were characteristic of acculturation, with the mean adoption (AD) score being significantly (P=.0002) higher for USVI-born persons than for African-Caribbean immigrants [AD=11.3 (95% CI=11.0-11.6) vs AD=10.6 (95% CI=10.4-10.8), respectively], while African Caribbean immigrants had a higher (P=.0001) traditional score (TS) compared to USVI-born persons [TS=9.0 (95% C=8.8-9.2) vs TS=7.0 (95% CI=6.7-7.2), respectively]. In regression analyses adjusting for age, education, income, smoking, and alcohol consumption, the 'adoption' factor was independently and positively related to BMI (P=.02) among USVI-born African Caribbeans, and to fasting glucose (P=.005) among African-Caribbean immigrants. CONCLUSIONS: Acculturation is associated with increased body size and diabetes risk in African Caribbeans in the USVI and appears to bear a differential impact according to place of birth. PMID- 12723009 TI - Advanced diabetic nephropathy disproportionately affects African-American females: cross-sectional analysis and determinants of renal survival in an academic renal clinic. AB - OBJECTIVE: The aims of this study were to study further the previously observed 2 fold higher prevalence of endstage renal disease (ESRD) due to diabetic nephropathy (DN) in African-American (AA) women, when compared to AA men, and to identify factors predictive of renal survival in this population. DESIGN: Cross sectional analysis of DN in an academic renal clinic. METHODS: We identified and retrospectively studied all patients with diabetic nephropathy enrolled in our renal clinic in 1996. Charts were reviewed for basic demographic factors, medications, and renal function. RESULTS: Of approximately 600 total patients, 141 were diagnosed with DN, and sufficient data was available on 119 (98 AA) of these, with a mean followup of 105 weeks. Patients were hypertensive, had advanced renal disease at presentation (creatinine [Cr]=3.97 mg/dL), and had been diabetic for 16.7 years. The AAs were disproportionately represented by females (69 vs 29). Seventy patients reached ESRD at the time of analysis (April 2000) with a mean time to dialysis of 79.3 weeks. Significant determinants for longer time to ESRD (TTE) were: lower presenting Cr, female gender, and AA race. A MAP<100 mm Hg over the course of followup was adversely related to TTE. Among AAs, taking an angiotensin converting enzyme (ACE) inhibitor, or a calcium channel blocker, was associated with better renal survival. Changes in renal function at 1 and 2 years were associated with age, urine protein at presentation, and MAP over the course of followup. CONCLUSIONS: In Mississippi, DN is more much more prevalent in AA females, compared to AA males. In those with advanced DN, MAP<100 mm Hg may lessen the chances for renal survival. PMID- 12723010 TI - Obesity, diabetes, hypertension, and vegetarian status among Seventh-Day Adventists in Barbados: preliminary results. AB - A population-based sample of Seventh-Day Adventists was studied to determine the relationship between vegetarian status, body mass index (BMI), obesity, diabetes mellitus (DM), and hypertension, in order to gain a better understanding of factors influencing chronic diseases in Barbados. A systematic sampling from a random start technique was used to select participants for the study. A standard questionnaire was used to collect data on demographic and lifestyle characteristics, to record anthropometrics and blood pressure measurements, and to ascertain the hypertension and diabetes status of participants. The sample population consisted of 407 Barbadian Seventh-Day Adventists (SDAs), who ranged in age from 25 to 74 years. One hundred fifty-three (37.6%) participants were male, and 254 (62.4%) were female, and 43.5% were vegetarians. The prevalence rates of diabetes and hypertension were lower among long-term vegetarians, compared to non-vegetarians, and long-term vegetarians were, on average, leaner than non-vegetarians within the same cohort. A significant association was observed between a vegetarian diet and obesity (vegetarian by definition P=.04, self-reported vegetarian P=.009) in this population. Other components of the study population lifestyle should be further analyzed to determine the roles they may plan in lessening the prevalence rates of obesity, diabetes, and hypertension. PMID- 12723011 TI - Personalized follow-up increases return rate at urban eye disease screening clinics for African Americans with diabetes: results of a randomized trial. AB - OBJECTIVE: To evaluate the effectiveness of using personalized follow-up, as compared to reminder letters, in increasing return rates at urban eye disease screening clinics for African Americans with diabetes, and to identify factors predictive of the patient's likelihood of returning for annual follow-up exams. RESEARCH DESIGN AND METHODS: All patients attending free community-based retinopathy screening clinics who were advised to return in one year for another diabetes eye evaluation (DEE) were randomized to standard or personalized follow up interventions. Patients in the standard follow-up group received reminder letters a month before it was time to return for their next annual DEE. Patients in the intensive personalized intervention also received the letters, but those patients who did not call for an appointment within 10 days received a phone call from project staff, encouraging them to return for a DEE. RESULTS: One hundred thirty-two African Americans with diabetes were randomized to one of the 2 treatments. The return rate for the intensive, personalized follow-up group was 66%, significantly (P=.001) higher than the 35% return rate for the standard follow-up group. CONCLUSION: This study demonstrated the efficacy of personal contact by telephone in improving return rates for annual DEEs in this population of patients. This finding is consistent with one of the key design principles of the project, which was to establish credible personal relationships with community leaders and patients as a means to maximize the utilization of the eye screening clinics. PMID- 12723012 TI - Racial/ethnic differences in the healthcare experience (coverage, utilization, and satisfaction) of US adults with diabetes. AB - OBJECTIVE: To examine racial/ethnic differences in healthcare coverage, utilization, and satisfaction, among US adults with diabetes. DESIGN AND SETTING: We conducted a cross-sectional analysis among 9443 adults with diabetes who participated in the 1999 Behavioral Risk Factor Surveillance System (BRFSS), a telephone survey of the civilian non-institutionalized US population aged > or = 18 yrs. MAIN OUTCOME MEASURES: We compared healthcare coverage, utilization, and satisfaction across 4 race/ethnicity categories: non-Hispanic Whites (NHW), non Hispanic Blacks (NHB), Hispanics (HSP), and others, and examined whether these factors were associated with self-rated health status. RESULTS: By self-report, more NHB (14.8%), HSP (20.7%), and members of other races (21.8%) were uninsured, compared to NHW (6.4%). Similarly, cost was a barrier to visiting a doctor for 23.9% of HSP, 19.5% of NHB, and 13.4% of members of other races; however, only 8.2% of non-Hispanic Whites reported cost as a barrier. More NHW (90.1%) and NHB (90.7) reported having had a check-up in the past year, compared to HSP (84.5%) or others (84.1%). All 3 variables exhibited significant differences by race or ethnicity (all P<.01). After adjustment for age, sex, income, education, and insulin use, the association with race/ethnicity persisted for health insurance coverage (P<.001), and for cost as a barrier (P<.003). Reporting cost as a barrier to visiting a doctor (P=.013), and rating one's overall health care as fair or poor (P=.001), were associated with poorer health status. CONCLUSIONS: These results suggest that ethnic minorities with diabetes report less healthcare coverage and more cost-related barriers to utilization, compared to non-Hispanic Whites. Persons with fair/poor health status were more likely to report cost barriers and poor satisfaction. Future research should focus on the reasons for such differences and on interventions to improve health care for minority populations. PMID- 12723013 TI - Gestational diabetes among Aboriginal Australians: prevalence, time trend, and comparisons with non-Aboriginal Australians. AB - OBJECTIVE: This paper investigates the prevalence, trends, and risk factors of gestational diabetes mellitus (GDM) in the Aboriginal population in Australia, and provides comparisons to the non-Aboriginal population. DESIGN: A retrospective population analysis of all deliveries (230,011) that took place in the state of South Australia between 1988 and 1999, inclusive. Data were obtained from the Pregnancy Outcome Unit of the Department of Human Services in South Australia, and include information on the demographical characteristics and medical status of both mothers and babies. MAIN OUTCOME MEASURES: Gestational diabetes mellitus prevalence rate among Aboriginal mothers, as compared to their non-Aboriginal counterparts. RESULTS: The age-standardized GDM rate for Aboriginal mothers was found to be more than 2.5 times higher than that for non Aboriginal mothers; there have been no significant changes, nor any decrease (as some previous studies have suggested), in this trend over the time period considered in this study. CONCLUSIONS: Clearly, the prevalence of GDM, however it is measured or defined, is still significantly higher in the Aboriginal population of Australia, as compared to the country's non-Aboriginal population. Researchers in the health field should target appropriate public health programs, such as universal screenings for GDM conditions, and health promotion procedures, toward the Aboriginal community. PMID- 12723014 TI - The relationship of spirituality and health outcomes in Black women with type 2 diabetes. AB - The purpose of this pilot study was to explore the relationships between spiritual well-being emotional distress, HbA1c values, and blood pressure levels in a convenience sample of 22 Black women with type 2 diabetes. Results revealed significant inverse correlations between diastolic blood pressure (BP) and both total spiritual well-being (r=-.51, P=.02) and religious well-being (RWB) (r= .55, P=.01). Women with higher RWB scores tended to have lower diastolic BP, as compared to their counterparts with lower RWB scores (z=2.78, P=.005). Emotional distress was positively related to systolic BP (r=.48, P=.03). These finding suggest that holistic care, addressing the spiritual and emotional dimensions, may foster improved BP levels among Black women with type 2 diabetes, thereby potentially reducing their high risk for secondary complications. PMID- 12723016 TI - Hypertriglyceridemic waist as a screening tool for CVD risk in indigenous Australian women. AB - Research has demonstrated that the simultaneous determination of waist circumference and fasting plasma triglyceride (TG) concentrations can identify men characterized by a metabolic triad of unconventional risk variables: increased levels of fasting insulin, apolipoprotein (apo) B, and a predominance of small, dense, low density lipoprotein (LDL) particles. The aim of this study was to assess the efficacy of using "hypertriglyceridemic waist" to identify individuals at high risk of CVD in a sample of indigenous Australian women, for whom 2 of the 3 non-traditional risk factors were measured (apo B and insulin). Subjects (N=80) were divided into subgroups on the basis of waist girth and TG levels. The TG/HDL ratio increased in women with both elevated waist (above 95 cm) and TG levels (above 2.0 mmol/L), who were also characterized by lower HDL and elevated LDL concentrations. Although there was no trend toward an increase in apo B with increasing waist girth and TG levels, apo B concentration was highest among subgroups with elevated waist and TG levels. Fasting insulin levels were higher with increasing waist girth, but not with increasing TG levels. Utilizing hypertriglyceridemic waist as a marker of high plasma insulin and apo B can be an important factor in assessing cardiovascular risk in indigenous Australian women, despite an unexpected apo B distribution. PMID- 12723015 TI - Adjunctive sympathoplegic therapy to ACE inhibition in Blacks with congestive heart failure: a comparison of alpha-1 with beta-1 blockade on exercise tolerance and cardiac sympathovagal reflex activity. AB - OBJECTIVES: Congestive heart failure (CHF) is characterized by an initial compensatory, but subsequently deleterious, activation of both the renin angiotensin (RAS) and the sympathetic nervous system (SNS). Incomplete suppression of the SNS may contribute to the residual mortality during optimal ACE inhibitor therapy in CHF. Carvedilol, a mixed alpha and beta-blocker with antioxidant properties, and other pure beta-adrenoceptor blockers reduce morbidity and mortality in Caucasians with CHF. However, beta-blocker monotherapy is of poor efficacy in Blacks with essential hypertension or in the treatment of glaucoma. The efficacy of beta-blockers in the treatment of African Americans with congestive heart failure is a controversial issue with conflicting findings. The aims of the present study were to examine and compare the cardiovascular, autonomic, and clinical effects of additional alpha-1, or beta-1 blockade in ACE inhibitor treated Black patients with moderate to severe CHF. METHODS: Twenty eight Nigerian patients with chronic CHF stabilized on digoxin and diuretics, were randomized to 3 groups of similar demographics according to a single blind, parallel group design. The patients were aged 53 +/- 6 years, and comprised 14 men and 14 women, with a mean cardiothoracic ratio of 0.66 +/- 0.03, and ejection fraction of 0.38 +/- 0.10, 60% hypertensive etiology. Group 1 patients received 5 mg enalapril alone, group 2 received 5 mg enalapril + 1 mg prazosin, and group 3 received 5 mg enalapril + 50 mg atenolol. All medication was taken daily for 4 weeks. Blood pressure, heart rate, pressure rate product, 6-minute walk test, NYHA class, and cardiac autonomic reflexes were measured at baseline and again at 2 and 4 weeks of treatment. Two-way repeated measures ANOVA, and a one-way ANOVA were used in data analysis. RESULTS: The 3 treatments caused significant (P<.001 ANOVA) and similar improvements for the NYHA class (-1.0 to -1.6), and increased the 6-minute distance covered (+130 m to +205 m). Although no treatment differences were observed, a trend suggesting a greater improvement with enalapril + atenolol became apparent. By the fourth week, the sympathoplegic treatments, enalapril + atenolol, and enalapril + prazosin, caused significant reductions in the pressure rate product (-3726 +/- 1885 mm Hg x beats/min; -3498 +/- 396 mm Hg beats/min, respectively), (compared to enalapril alone (-1349 +/- 894 mm Hg x beats/min) (P<.001 ANOVA). During the Valsalva maneuver, the phase IV bradycardia were significantly greater after treatment with enalapril + atenolol (944 +/- 66 msec) or with enalapril + prazosin (825 +/- 48 msec), compared to enalapril alone (760 +/- 45 msec) (P<.001 ANOVA). The phase II Valsalva tachycardia were similar between treatments. The respiratory sinus arrhythmia ratio increased significantly (P<.005 ANOVA) and equally on all treatments. However, the pressor and chronotropic responses to forearm isometric handgrip increased significantly on the enalapril + prazosin combination (P<.02), compared to the other treatments. CONCLUSIONS: Our findings demonstrated not only the safety of providing additional therapy with alpha-1 or beta-1 receptor blockade concurrent with ACE inhibition in Blacks with CHF, but also the resultant improvement in exercise tolerance and NYHA class. Compared to using ACE inhibition alone, the combined therapies caused a marked reduction in the pressure rate product, an index of myocardial oxygen consumption, and a greater enhancement of cardiac parasympathetic activity. Selective beta-1 blockade caused a greater enhancement of central baroreceptor vagal activity compared to alpha-1 blockade. Conversely, the pressor and chronotropic abnormalities during forearm isometric handgrip in CHF, were normalized by alpha-1, but not beta-1, blockade. Thus, the combined reflex cardiac vagal augmentation following selective beta-1 blockade, and the hemodynamic effects of alpha-1 antagonism with concurrent ACE inhibition, may be of major therapeutic and prognostic benefit in Blacks with non ischemic (hypertensive) CHF stabilized on digoxin and diuretics. PMID- 12723017 TI - Changes in prevalence of nonfatal coronary heart disease in the United States from 1971-1994. AB - OBJECTIVE: To examine temporal trends in the prevalence of nonfatal coronary heart disease in the United States. DESIGN: Four national cross-sectional health surveys: National Health and Nutrition Examination Survey (NHANES) I (1971-1975), NHANES II (1976-1980), NHANES III (1988-1994), and Hispanic HANES (HHANES) (1982 1984). SETTING: United States. PARTICIPANTS: Persons aged 40-74 years. MAIN OUTCOME MEASUREMENTS: Prevalence of angina, self-reported myocardial infarction, and electrocardiographically defined myocardial infarction (ECG-MI). RESULTS: Generally, the age-adjusted prevalence of angina pectoris was higher among women than men, but the reverse was true for self-reported myocardial infarction and ECG-MI. Increases in the prevalence of angina pectoris occurred for Mexican American men and women, and African-American women, but were not statistically significant for the latter. Age-adjusted rates of self-reported myocardial infarction increased among African-American men (P=.019) and women (P=.005) and Mexican-American men (P>.05), but decreased among White men (P>.05) and women (P>.05). The prevalence of age-adjusted ECG-MI decreased among African-American men and women, White women, and to a lesser degree, White men; however, none of these decreases were statistically significant. Relative standard errors for ECG MI prevalence in NHANES I and II among African Americans were large; therefore, prevalence trends need to be interpreted cautiously. CONCLUSIONS: The decreases in ECG-MI could be due either to decreased incidence of coronary heart disease or myocardial infarction, or increases in the rates of timely cardiac interventions that minimize damage to the myocardium. PMID- 12723019 TI - Factors associated with overweight/obesity in economically active South African populations. AB - OBJECTIVE: To investigate factors associated with self-reported weight status of economically active adults from the 4 major ethnic groups in South Africa. DESIGN: Cross-sectional survey. SETTING: South Africa. PARTICIPANTS: A random sample (N=2100) was selected from a database of economically active adults from the 4 major ethnic groups in South Africa (Black, White, Asian, and mixed ancestry). Of the 2100 selected, 554 subjects returned mailed questionnaires. MAIN OUTCOME MEASURES: Weight status (BMI), dieting history, meal patterns, intake of high fat food items and alcohol, level of physical activity, smoking habits, family history of obesity, and socioeconomic characteristics. RESULTS: Identified risk factors for overweight/obesity included: Black ethnicity, education level < or = Grade 7, inactivity, and at least one overweight parent. Protective factors included: describing one's own weight as under- or normal weight, hardly ever or never binging, not having tried to lose weight during the past year, and describing one's own health as excellent. Factors that were not related to overweight/obesity in this group included: employment status, income, smoking, meal patterns, intake of high fat food items and alcohol, and "sick" days taken off from work during the prior 6 months. PMID- 12723020 TI - Retention factors for participants of an inner-city community-based asthma intervention study. AB - Participant retention is a significant challenge for asthma field trials examining the effectiveness of prevention strategies in inner-city communities. Here, the authors evaluate factors associated with participant retention in an inner-city, pediatric, asthma intervention trial in Atlanta, Georgia, during 1998 2000. Demographic, clinical, residential, personnel, and logistical variables were analyzed by chi-square and Wilcoxon rank sum nonparametric tests to compare children who remained in the asthma study with those who were dropped. Of the 489 participants, 486 (99%) were African-American, 467 (96%) were non-Hispanic, 281 (57%) were male, and 142 (29%) remained in the study. Of the 347 dropouts, 149 (43%) were dropped because of missing study visits. Retention rates were significantly higher (P<.05) for participants enrolled in the second year of the study (2nd yr=43%, 1st yr=19%), for those who lived longer at the same residence (> or = 3 yrs=36%, 2-3 yrs=26%, 1-<2 yr=22%), and for those enrolled during a face-to-face follow-up home visit, rather than at the emergency department (ED) (follow-up=38%, ED=27%). Neither sex nor enrollment season were associated with retention. These findings underscore the importance of performing a comprehensive pilot study and considering a home residency period for participant enrollment eligibility, along with alternative study methods that take into account the challenges of retaining participants. PMID- 12723018 TI - Migration history, health behaviors, and cardiovascular disease risk factors in overweight Mexican-American women. AB - PURPOSE: This research examined whether the migration history of overweight Mexican-American women had an independent effect on cardiovascular risk factors, or whether it was mediated by health behavior changes. DATA AND METHODS: Cross sectional data from 390 overweight, non-diabetic Mexican-American women (aged 18 to 65 years), all recruited from Starr County, Texas, were used for this analysis. Migration history was inferred from birthplaces of subjects and relatives, and length of residence in the United States. Health behaviors included tobacco and alcohol use, sleeping, exercise, and dietary practices. The cardiovascular disease risk factor variables (CDRFVs) studied were plasma glucose, abdominal obesity, blood pressures, and blood lipids. A migration history score (MHS) was developed from factor analysis, almost equally contributed to by the 9 migration history variables. Healthy habits were defined by 6 variables, and 3 factors (blood pressures, lipids/glucose, and body fat/glucose) were used for the CDRFVs. FINDINGS AND CONCLUSION: MHS was correlated positively with socioeconomic status, and negatively with family stress. Older women had healthier drinking and sleeping habits. Women with a higher migration history score exhibited poorer exercise habits, and increased blood pressures. After adjusting for the effect of healthy exercise habits on blood pressures, the impact of migration history on blood pressures became non significant (P>.05), leading to the conclusion that healthy exercise behaviors mediated the negative relationship of MHS with blood pressures. Age was independently positively correlated with all CDRFVs. Age also weakly moderated the negative relationship of MHS and healthy exercise habits. PMID- 12723021 TI - Data quality and adjusted Hispanic mortality in the United States, 1989-1991. AB - OBJECTIVES: Hispanics appear to live longer compared to other ethnic groups in the United States. Our main objective was to determine whether data quality biases mortality statistics. We calculated the impact of misclassification of ethnicity on death certificates in order to create adjusted mortality estimates. METHODS: We used the National Mortality Follow-Back Survey of 1993 (NMFS) for our assessment of ethnicity misclassification. We then created misclassification estimates for 10-year age-sex groups, and used these to correct mortality estimates for 1989-1991. RESULTS: The overall predictive value positive (PV+) and sensitivity were 0.981 and 0.805, respectively, for men; and 0.994 and 0.902, respectively, for women. Age-specific adjustment for misclassification on death rates caused Hispanic male life expectancy to go from a 1.01 year advantage over White non-Hispanics, to a 1.83 year deficit, with a similar pattern found for females. CONCLUSIONS: The apparent Hispanic advantage in life expectancy is influenced by misclassification of ethnicity. Misclassification of ethnicity on death certificates biases Hispanic mortality downward, thereby falsely inflating life expectancy. PMID- 12723022 TI - For the patient. Customs and beliefs can affect your health. Cultural orientation and diabetes self-care in low-income African-Americans with type 2 diabetes mellitus. PMID- 12723023 TI - For the patient. The effectiveness of a community health worker outreach program on healthcare utilization of west Baltimore City Medicaid patients with diabetes, with or without hypertension. PMID- 12723024 TI - For the patient. African-American women at risk of diabetes and kidney disease. Advanced diabetic nephropathy disproportionately affects African-American females: cross-sectional analysis and determinants of renal survival in an academic renal clinic. PMID- 12723025 TI - For the patient. Are vegetarians at less risk for obesity, diabetes, and hypertension? Obesity, diabetes, hypertension, and vegetarian status among Seventh-Day Adventists in Barbados: preliminary results. PMID- 12723026 TI - For the patient. Eye screening can prevent eye disease. Personalized follow-up increases return rate at urban eye disease screening clinics for African Americans with diabetes: results of a randomized trial. PMID- 12723028 TI - For the patient. Hypertriglyceridemic waist as a screening tool for CVD risk in Australian indigenous women. PMID- 12723027 TI - For the patient. New treatments for heart failure in Black people. Adjunctive sympathoplegic therapy to ACE inhibition in Blacks with congestive heart failure: a comparison of alpha-1 with beta-1 blockade on exercise tolerance and cardiac sympathovagal reflex activity. PMID- 12723029 TI - For the patient. The importance of health behaviors for better heart health. Migration history, health behaviors, and cardiovascular disease risk factors in overweight Mexican-American women. PMID- 12723030 TI - For the patient. What lifestyle choices place people at-risk of obesity? Factors associated with overweight/obesity in economically active South African populations. PMID- 12723031 TI - For the patient. Racial/ethnic differences in the health care experience of US adults with diabetes. PMID- 12723032 TI - For the patient. Do Hispanics really live longer or is ethnicity reported incorrectly on death certificates? Data quality and adjusted Hispanic mortality in the US, 1989-1991. PMID- 12723033 TI - An expression cDNA library for suppression cloning in yeast mutants, complementation of a yeast his4 mutant, and EST analysis from the symbiotic basidiomycete Hebeloma cylindrosporum. AB - An oriented expression library was constructed from the mycelia of the symbiotic model fungus Hebeloma cylindrosporum in the high-level yeast expression vector pDR196. DNA sequencing of approximately 500 expressed sequence tags (ESTs) showed that 15% correspond to known genes, two thirds contain sequences with unknown function, andthe remaining 20% showed no significant similarity to any known genes. The ESTs had a GC content between 44 and 56%, with most of them having a GC content of 52-54%, which could be correlated with GC contents of fungal genes. The library was successfully used to identify the Hebeloma HIS4 gene by functional complementation of a yeast his4 mutant. Thus, the library may serve as a powerful tool for identification and characterization of mycorrhizal genes by EST analysis and for the identification of ectomycorrhizal genes by means of suppression cloning. PMID- 12723034 TI - The genetic basis of C-glycosyl flavone B-ring modification in maize (Zea mays L.) silks. AB - Resistance to corn earworm (CEW) (Helicoverpa zea Boddie) has been attributed to high concentrations of C-glycosyl flavones and chlorogenic acid in maize (Zea mays L.) silks. The most common C-glycosyl flavones isolated from maize silks are maysin, apimaysin, and methoxymaysin, which are distinguished by their B-ring substitutions. For a better understanding of the genetic mechanisms underlying the synthesis of these compounds, we conducted a quantitative trait locus (QTL) study with two populations: (Tx501 x NC7A)F2 and (Tx501 x Mp708)F2. For chlorogenic acid, maysin, and methoxymaysin concentration, the major QTL for both populations was located on chromosome 4 near umc1963. For apimaysin, the major QTL in both populations was located at the position of the pr1 locus on chromosome 5. The QTL alleles on chromosome 4 that increased the synthesis of methoxymaysin significantly decreased the synthesis of maysin and chlorogenic acid. This decrease in maysin concentration was four-fold greater than the increase in methoxymaysin. Our results indicate that the QTL on chromosome 4, responsible for the increase in methoxymaysin synthesis, alters the dynamics of both the phenylpropanoid and flavonoid pathways. PMID- 12723035 TI - A new karyological variant of Oecomys (Rodentia: Sigmodontinae) and its phylogenetic relationship based on molecular data. AB - A new karyological variant of the Sigmodontinae genus Oecomys was detected in specimens from Brazilian Pantanal. Karyologic analyses showed 2n = 72 and FN = 90, which differs from all known chromosomal complements of the genus. This new karyomorph showed distinctive morphological attributes and could not be attributed to any described Oecomys species. Molecular analysis using cytochrome b sequence data suggested the monophyly of Oecomys. Maximum-likelihood analyses placed the Oecomys sp. haplotype clade as the sister branch to Oecomys bicolor, whereas maximum-parsimony analyses did not resolve its relationship with the other Oecomys species. Kimura's two parameters distance estimates between this and other Oecomys species are high, and active gene flow has been found not to coexist with these divergence estimates. PMID- 12723036 TI - Mapping QTL for yield components, vigor, and resistance to Phytophthora palmivora in Theobroma cacao L. AB - Quantitative trait loci (QTL) mapping for agronomic traits was carried out in cocoa (Theobroma cacao L.). Regions of the genome involved in yield, vigor, and resistance to Phytophthora palmivora were identified. Three heterozygous clones, one upper Amazon Forastero (IMC78) and two Trinitario (DR1 and S52), were crossed with the same male parent, a lower Amazon Forastero (Catongo), known to be highly homozygous. Observations were made on progeny over nine consecutive years. One to three QTL related to yield were detected in each of the three populations, located on chromosomes 1, 2, 4, 5, 9, and 10. They explained between 8.1 and 19.3% of the phenotypic variation and showed various levels of repeatability. In IMC78, the QTL detected on chromosome 5 was the most repeatable over years. The QTL for the average individual pod weight on chromosome 4 was the most significant with an LOD of 17.3 and an R2 of 43.7. QTL related to these traits were identified in the same region of the genome in clones of different genetic groups. This suggests that molecular markers can be used to improve cocoa varieties. PMID- 12723038 TI - Genetic dissection of partial resistance to race 6 of Venturia inaequalis in apple. AB - Scab, caused by the fungus Venturia inaequalis, is one of the most important diseases of apple (Malus x domestica). The major resistance gene, Vf, has been widely used in apple breeding programs, but two new races of the fungus (races 6 and 7) are able to overcome this gene. A mapped F1 progeny derived from a cross between the cultivars Prima and Fiesta has bee n inoculated with two monoconidial strains of race 6. These strains originated from sporulating leaves of 'Prima' and a descendant of 'Prima' that were grown in an orchard in northern Germany. 'Prima' carries the Vf resistance gene, whereas 'Fiesta' lacks Vf. A large variation in resistance and (or) susceptibility was observed among the individuals of the progeny. Several quantitative trait loci (QTLs) for resistance were identified that mapped on four genomic regions. One of them was located in the very close vicinity of the Vf resistance gene on linkage group LG-1 of the 'Prima' genetic map. This QTL is isolate specific because it was only detected with one of the two isolates. Two out of the three other genomic regions were identified with both isolates (LG-11 and LG-17). On LG-11, a QTL effect was detected in both parents. The genetic dissection of this QTL indicated a favourable intra-locus interaction between some parental alleles. PMID- 12723037 TI - Evolutionary dynamics of a satellite DNA in the tiger beetle species pair Cicindela campestris and C. maroccana. AB - Satellite repeat elements are an abundant component of eukaryotic genomes, but not enough is known about their evolutionary dynamics and their involvement in karyotype and species differentiation. We report the nucleotide sequence, chromosomal localization, and evolutionary dynamics of a repetitive DNA element of the tiger beetle species pair Cicindela maroccana and Cicindela campestris. The element was detected after restriction digest of C. maroccana total genomic DNA with EcoRI as a single band and its multimers on agarose gels. Cloning and sequencing of several isolates revealed a consensus sequence of 383 bp with no internal repeat structure and no detectable similarity to any entry in GenBank. Hybridization of the satellite unit to C. maroccana mitotic and meiotic chromosomes revealed the presence of this repetitive DNA in the centromeres of all chromosomes except the Y chromosome, which exhibited only a very weak signal in its short arm. PCR-based tests for this satellite in related species revealed its presence in the sister species C. campestris, but not in other closely related species. Phylogenetic analysis of PCR products revealed well-supported clades that generally separate copies from each species. Because both species exhibit the multiple X chromosome karyotypic system common to Cicindela, but differ in their X chromosome numbers (four in C. maroccana vs. three in C. campestris), structural differences could also be investigated with regard to the position of satellites in a newly arisen X chromosome. We find the satellite in a centromeric position in all X chromosomes of C. maroccana, suggesting that the origin of the additional X chromosome involves multiple karyotypic rearrangements. PMID- 12723039 TI - Fine mapping of quantitative trait loci for improved fruit characteristics from Lycopersicon chmielewskii chromosome 1. AB - The near-isogenic line (NIL) TA1150 contains a 56-cM introgression from Lycopersicon chmielewskii chromosome 1 and has several interesting phenotypic characteristics including fruit with orange color, high levels of soluble solids, thick pericarp, small stem scars, and good firmness. A set of overlapping recombinant lines (subNILs) was developed and field tested to fine map the quantitative trait loci (QTL) controlling these traits. The results indicated that the solids, pericarp thickness, and firmness QTL are distinct from the color locus. Several of the QTL mapped in this study, including the soluble-solids QTL, probably correspond to QTL mapped in other wild species of tomato. However, analysis of a set of TA523 subNILs containing complementary introgressions from Lycopesicon hirsutum chromosome 1 suggests that this wild species may contain a different locus for improved soluble solids. Thus, it might be possible to combine the L. chmielewskii and L. hirsutum alleles for these loci in a single line with the potential for extremely highly soluble solids. The TA1150 subNIL TA1688 contains the smallest introgression of the solids locus (approximately 19 cM), as well as the pericarp thickness and firmness QTL, with a yield that was equivalent to two of the three control lines. Isolation of recombinant subNILs from TA1688 should break the linkage between orange color and high solids and provide a small introgressed segment for marker-assisted breeding and genetic improvement of processing tomato. PMID- 12723040 TI - Genetic diversity of different apricot geographical groups determined by SSR markers. AB - Forty apricot cultivars with different geographic origins belonging to the germplasm collections of St. Istvan University (Budapest, Hungary) and the Instituto Valenciano de Investigaciones Agrarias (IVIA) (Valencia, Spain) were studied by means of SSR markers. The aim of the study was to determine the genetic relationships among genotypes from different eco-geographical groups. Sixteen primer pairs flanking microsatellite sequences in the peach genome were assayed. Eleven of them were polymorphic in the set of cultivars studied and allowed every genotype to be unambiguously distinguished. Genetic diversity in the population studied was analyzed using several variability parameters. A total of 34 alleles were detected with a mean value of 3.1 alleles/locus. The expected heterozygosity mean was 0.46 and the observed heterozygosity was 32% on an average leading to a high value of the Wright's fixation index (0.32). Additionally, UPGMA cluster analysis based on Nei's genetic distance grouped genotypes according to their geographic origins and pedigrees. SSR markers have proved to be an efficient tool for fingerprinting cultivars and conducting genetic-diversity studies in apricot. PMID- 12723041 TI - Genetic mapping of the Sorghum bicolor vp1 gene and its relationship with preharvest sprouting resistance. AB - The plant vp1 gene encodes a transcription factor originally identified in maize that participates in the control of the transition from embryogenesis to seed germination. Different lines of evidence suggest that vp1 participates in preharvest sprouting (PHS) resistance in cereals, but the genetic relationship is not yet established. Thus, the aim of this study was to investigate if there is a connection between vp1 and the formerly documented quantitative trait loci for PHS in Sorghum bicolor. This gene was mapped in sorghum using two well-studied inbred lines displaying contrasting PHS phenotypes. Linkage analysis revealed that the S. bicolor vp1 (Sbvp1) locus is linked to markers located on chromosomes 3 and 8 in maize. Analysis indicated that this gene is not correlated with PHS at a statistically significant level. Structural analysis of the Sbvp1 gene from both inbred lines showed a high degree of overall conservation (low polymorphism), even in the promoter region. These findings, taken together with the previously observed correlation between the pattern of expression of this gene and PHS performance, suggest that Sbvp1 might act downstream in the signalling pathway that leads to seed germination, or even be differentially regulated as a consequence rather than as a cause of the phenotypic behaviour. PMID- 12723042 TI - Predicting progeny performance in common bean (Phaseolus vulgaris L.) using molecular marker-based cluster analysis. AB - Recovery of superior individuals from a cross based solely on the phenotypic characteristics ofsingle-plant selections is inefficient because some traits, like yield, have low heritabilities, or because it is difficult to create the correct conditions for selection, as with disease resistance. In contrast, molecular markers are highly heritable and unaffected by environmental conditions. The objective of this study was to investigate the potential of molecular markers to identify superior lines in a breeding population by examining relationships between genetic distances (GDs) and phenotypic data for eight agronomic and architectural traits (branch angle, height, hypocotyl diameter, lodging, maturity, upper pods, pods per plant, and yield) obtained from three locations over a two-year period. From an elite common bean (Phaseolus vulgaris L.) cross, 110 recombinant inbred lines (RILs) and the two parents were screened with 116 random amplified polymorphic DNA (RAPD) markers. Pairwise GD values were calculated between each line and a selected "target" (the parent 'OAC Speedvale') using the Jaccard method and correlated to the trait data. The correlations were low and non-significant for all traits, except for branch angle (r = 0.30), maturity (r = -0.25), and pods per plant (r = 0.35). The lines were also grouped according to their cluster-based GD from the target parent using UPGMA cluster analysis. Trait data of lines within groups were combined and correlated to cluster-based GD. Correlation values were large and significant for all traits. Additionally, one-half of the top 10 yielding lines and nearly one third of the best phenotypically ranked lines were present within the 13% of lines clustered nearest the target. A selection method using marker-based cluster analysis (MBCA) is suggested to assist phenotypic selection by directing a breeder's attention to a subsample of the population containing a high proportion of superior lines. PMID- 12723043 TI - Comparison of peach and Arabidopsis genomic sequences: fragmentary conservation of gene neighborhoods. AB - We examined the degree of conservation of gene order in two plant species, Prunus persica (peach) and Arabidopsis thaliana (thale cress), whose lineages diverged more than 90 million years ago. In the three peach genomic regions studied, segments with a gene order congruent with A. thaliana were short (two to three genes in length); and for any peach region, corresponding segments were found in diverse locations in the A. thaliana genome. At the gene level and lower, the A. thaliana sequence was enormously useful for identifying likely coding regions in peach sequences and in determining their intron-exon structure. The peach BAC sequence data reported here contained a BLAST-detectable putative coding sequence an average of every 7 kb, and the peach introns identified in this study were, on average, almost twice the length of the corresponding introns in A. thaliana. PMID- 12723044 TI - Development and characterization of EST-derived simple sequence repeat (SSR) markers for pasture grass endophytes. AB - Fungal endophytes of the genus Neotyphodium are common in temperate pasture grass species and confer both beneficial and deleterious agronomic characteristics to their hosts. The aim of this study was to develop molecular markers based on simple sequence repeat (SSR) loci for the identification and assessment of genetic diversity among Neotyphodium endophytes in grasses. Expressed sequence tags (ESTs) from both Neptyphodium coenophialum and Neotyphodium lolii were examined, and unique SSR loci were identified in 9.7% of the N. coenophialum sequences and 6.3% of the N. lolii sequences. A variety of SSRs were present, although perfect trinucleotide repeat arrays were the most common. Primers were designed to 50 SSR loci from N. coenophialum and 57 SSR loci from N. lolii and were evaluated using 20 Neotyphodium and Epichloe isolates. A high proportion of the N. coenophialum and N. lolii primers produced amplification products from the majority of isolates and most of these primers detected genetic variation. SSR markers from both N. coenophialum and N. lolii detected high levels of polymorphism between Neotyphodium and Epichloe species, and low levels of polymorphism within N. coenophialum and N. lolii. SSR markers may be used in appropriate combinations to discriminate between species. Comparison with amplified fragment length polymorphism (AFLP) data demonstrated that the SSR markers were informative for the assessment of genetic variation within and between endophyte species. These markers may be used to identify endophyte taxa and to evaluate intraspecific population diversity, which may be correlated with variation for endophyte-derived agronomic traits. PMID- 12723045 TI - Patterns of genome duplication within the Brassica napus genome. AB - The progenitor diploid genomes (A and C) of the amphidiploid Brassica napus are extensively duplicated with 73% of genomic clones detecting two or more duplicate sequences within each of the diploid genomes. This comprehensive duplication of loci is to be expected in a species that has evolved through a polyploid ancestor. The majority of the duplicate loci within each of the diploid genomes were found in distinct linkage groups as collinear blocks of linked loci, some of which had undergone a variety of rearrangements subsequent to duplication, including inversions and translocations. A number of identical rearrangements were observed in the two diploid genomes, suggesting they had occurred before the divergence of the two species. A number of linkage groups displayed an organization consistent with centric fusion and (or) fission, suggesting this mechanism may have played a role in the evolution of Brassica genomes. For almost every genetically mapped locus detected in the A genome a homologous locus was found in the C genome; the collinear arrangement of these homologous markers allowed the primary regions of homoeology between the two genomes to be identified. At least 16 gross chromosomal rearrangements differentiated the two diploid genomes during their divergence from a common ancestor. PMID- 12723046 TI - Segmental allotetraploidy and allelic interactions in buffelgrass (Pennisetum ciliare (L.) Link syn. Cenchrus ciliaris L.) as revealed by genome mapping. AB - Linkage analyses increasingly complement cytological and traditional plant breeding techniques by providing valuable information regarding genome organization and transmission genetics of complex polyploid species. This study reports a genome map of buffelgrass (Pennisetum ciliare (L.) Link syn. Cenchrus ciliaris L.). Maternal and paternal maps were constructed with restriction fragment length polymorphisms (RFLPs) segregating in 87 F1 progeny from an intraspecific cross between two heterozygous genotypes. A survey of 862 heterologous cDNAs and gDNAs from across the Poaceae, as well as 443 buffelgrass cDNAs, yielded 100 and 360 polymorphic probes, respectively. The maternal map included 322 RFLPs, 47 linkage groups, and 3464 cM, whereas the paternal map contained 245 RFLPs, 42 linkage groups, and 2757 cM. Approximately 70 to 80% of the buffelgrass genome was covered, and the average marker spacing was 10.8 and 11.3 cM on the respective maps. Preferential pairing was indicated between many linkage groups, which supports cytological reports that buffelgrass is a segmental allotetraploid. More preferential pairing (disomy) was found in the maternal than paternal parent across linkage groups (55 vs. 38%) and loci (48 vs. 15%). Comparison of interval lengths in 15 allelic bridges indicated significantly less meiotic recombination in paternal gametes. Allelic interactions were detected in four regions of the maternal map and were absent in the paternal map. PMID- 12723047 TI - Molecular cytogenetic analysis of wheat-barley hybrids using genomic in situ hybridization and barley microsatellite markers. AB - In the present investigation, genomic in situ hybridization (GISH) and barley microsatellite markers were used to analyse the genome constitution of wheat barley hybrids from two backcross generations (BC1 and BC2). Two BC1 plants carried 3 and 6 barley chromosomes, respectively, according to GISH data. Additional chromosomal fragments were detected using microsatellites. Five BC2 plants possessed complete barley chromosomes or chromosome segments and six BC2 plants did not preserve barley genetic material. Molecular markers revealed segments of the barley genome with the size of one marker only, which probably resulted from recombination between wheat and barley chromosomes. The screening of backcrossed populations from intergeneric hybrids could be effectively conducted using both genomic in situ hybridization and molecular microsatellite markers. GISH images presented a general overview of the genome constitution of the hybrid plants, while microsatellite analysis revealed the genetic identity of the alien chromosomes and chromosomal segments introgressed. These methods were complementary and provided comprehensive information about the genomic constitution of the plants produced. PMID- 12723049 TI - The organization of genes tightly linked to the Ha locus in Aegilops tauschii, the D-genome donor to wheat. AB - The grain hardness locus, Ha, is located at the distal end of the short arm of chromosome 5D in wheat. Three polypeptides, puroindoline-a, puroindoline-b, and grain softness protein (GSP-1), have been identified as components of friabilin, a biochemical marker for grain softness, and the genes for these polypeptides are known to be tightly linked to the Ha locus. However, this region of the chromosome 5D has not been well characterized and the physical distance between the markers is not known. Separate lambda clones containing the puroindoline-a gene and the puroindoline-b gene have been isolated from an Aegilops tauschii (the donor of the D genome to wheat) genomic lambda library and investigated. Considerable variation appears to exist in the organization of the region upstream of the gene for puroindoline-b among species closely related to wheat. Using in situ hybridization the genes for puroindoline-a, -b, and GSP-1 were demonstrated to be physically located at the tip of the short arm of chromosome 5 of A. tauschii. Four overlapping clones were isolated from a large-insert BAC library constructed from A. tauschii and of these one contained genes for all of puroindoline-a, puroindoline-b, and GSP-1. The gene for puroindoline-a is located between the other two genes at a distance no greater than approximately 30 kb from either gene. The BAC clone containing all three known genes was used to screen a cDNA library constructed from hexaploid wheat and cDNAs that could encode novel polypeptides were isolated. PMID- 12723048 TI - Shoot meristem: an ideal explant for Zea mays L. transformation. AB - We report on a rapid high-frequency somatic embryogenesis and plant regeneration protocol for Zea mays. Maize plants were regenerated from complete shoot meristem (3-4 mm) explants via organogenesis and somatic embryogenesis. In organogenesis, the shoot meristems were directly cultured on a high-cytokinin medium comprising 5-10 mg x L(-1) 6-benzylaminopurine (BAP). The number of multiple shoots produced per meristem varied from six to eight Plantlet regeneration through organogenesis resulted in just four weeks. Callus was induced in five days of incubation on an auxin-modified Murashige and Skoog (MS) medium. Prolific callus, with numerous somatic embryos, developed within 3-4 weeks when cultured on an auxin medium containing 5 mg 2,4-dichlorophenoxyacetic acid x L(-1). The number of multiple shoots varied from three to six per callus. Using R23 (Pioneer, Hi-Bred, Johnston, Iowa), the frequency of callus induction was consistently in excess of 80% and plant regeneration ranged between 47 and 64%. All regenerated plantlets survived in the greenhouse and produced normal plants. Each transgenic plant produced leaves, glumes, and anthers that uniformly expressed green fluorescent protein (GFP). The GFP gene segregated in the pollen. Based on this data it is concluded that the transgenics arose from single-cell somatic embryos. The rate of transfer DNA (T-DNA) transfer to complete shoot meristems of Zea mays was high on the auxin medium and was independent of using super-virulent strains of Agrobacterium. PMID- 12723050 TI - Moth sex chromatin probed by comparative genomic hybridization (CGH). AB - Abstract: Comparative genomic hybridization (CGH) with a probe mixture of differently labeled genomic DNA from females and males highlighted the W chromosomes in mitotic plates and the W chromatin in polyploid interphase nuclei of the silkworm Bombyx mori, the flour moth Ephestia kuehniella, and the wax moth Galleria mellonella. The overproportionate fluorescence signal indicated an accumulation of repetitive sequences in the respective W chromosomes. Measurements of the fluorescence signals revealed two components, one that is present also in male DNA (non-W chromosomes) and another one that is present only in or preponderantly in female DNA (W chromosomes). While the W chromosomes of E. kuehniella and G. mellonella had both components, that of B. mori appeared to lack the latter component. Our results show that CGH can be applied to obtain a first estimate of the sequence composition of sex chromosomes in species from which otherwise little is known on the molecular level. PMID- 12723051 TI - Membrane current from transmembrane potentials in complex core-conductor models. AB - Core-conductor models, used to integrate the behavior of the longitudinal currents with the distributed voltages of electrically active tissue, have evolved for over a century. A critical step in the use of such models is the computation of membrane current from the set of distributed transmembrane potential values that exist at a given moment, where the potentials are obtained either experimentally or computationally. Over time, interest has developed in a number of substantial extensions of the original model to include such features as nonuniform spatial resistances, loop instead of linear structure, and multiple sites of extracellular stimulation. This paper concisely restates and extends the equations for calculation of transmembrane currents with the systematic inclusion of alternative cases, noting how they reduce to the standard forms. An important issue is how complex the calculation of membrane current has to be. Thus, the paper goes on to show criteria (based on the uniformity of resistance and the presence of stimulation) for deciding when membrane currents can be obtained with a relatively simple calculation with a single equation involving local variables versus with a more complex calculation involving the simultaneous solution of a (possibly large) set of equations. PMID- 12723052 TI - A numerical scheme for modeling wavefront propagation on a monolayer of arbitrary geometry. AB - The majority of models of wavefront propagation in cardiac tissue have assumed relatively simple geometries. Extensions to complicated three-dimensional (3-D) representations are computationally challenging due to issues related both to problem size and to the correct implementation of flux conservation. In this paper, we present a generalized finite difference scheme (GDFS) to simulate the reaction-diffusion system on a 3-D monolayer of arbitrary shape. GDFS is a vertex centered variant of the finite-volume method that ensures local flux conservation. Owing to an effectively lower dimensionality, the overall computation time is reduced compared to full 3-D models at the same spatial resolution. We present the theoretical background to compute both the wavefront conduction and local electrograms using a matrix formulation. The same matrix is used for both these quantities. We then give some results of simulation for simple monolayers and complex monolayers resembling a human atria. PMID- 12723053 TI - Method for unsupervised classification of multiunit neural signal recording under low signal-to-noise ratio. AB - Neural spike sorting is an indispensable step in the analysis of multiunit extracellular neural signal recording. The applicability of spike sorting systems has been limited, mainly to the recording of sufficiently high signal-to-noise ratios, or to the cases where supervised classification can be utilized. We present a novel unsupervised method that shows satisfactory performance even under high background noise. The system consists of an efficient spike detector, a feature extractor that utilizes projection pursuit based on negentropy maximization (Huber, 1985 and Hyvarinen et al, 1999), and an unsupervised classifier based on probability density modeling using mixture of Gaussians (Jain et al., 2000). Our classifier is based on the mixture model with a roughly approximated number of Gaussians and subsequent mode-seeking. It does not require accurate estimation of the number of units present in the recording and, thus, is better suited for use in fully automated systems. The feature extraction stage leads to better performance than those utilizing principal component analysis and two nonlinear mappings for the recordings from the somatosensory cortex of rat and the abdominal ganglion of Aplysia. The classification method yielded correct classification ratio as high as 95%, for data where it was only 66% when a kappa means-type algorithm was used for the classification stage. PMID- 12723054 TI - Reduction of interference in oscillometric arterial blood pressure measurement using fuzzy logic. AB - In oscillometry, oscillation amplitudes (OAs) embedded in the cuff pressure are drastically affected by a variety of artifacts and cardiovascular diseases, leading to inaccurate arterial blood pressure (ABP) measurement. The purpose of this paper is to improve the accuracy in the arterial pressure measurement by reducing interference in the OAs using a recursive weighted regression algorithm (RWRA). This method includes a fuzzy logic discriminator (FLD) and a recursive regression algorithm. The FLD is used to reduce the effect of artifacts caused by measurement motion disturbance or cardiovascular diseases, and to determine the truthfulness of the oscillation pulse. According to the truth degree, the relationship between the cuff pressure and OA is reconstructed using the regression algorithm. Because the regression method must utilize inverse matrix operation, which will be difficult to implement in an automatic or ambulatory monitor, the recursive regression method is proposed to solve this problem. To test the performance of this RWRA, 47 subjects underwent the ABP measurement using both the auscultation and the oscillometry combined with the RWRA. It was found that the average difference between the pooled blood pressures measured by the auscultation and those by the oscillometry combined with the RWRA was found to be only 4.9 mmHg. Clinical results demonstrated that the proposed RWRA is more robust than the traditional curve fitting algorithm (TCFA). We conclude that the proposed RWRA can be applied to effectively improve the accuracy of the oscillometric blood pressure measurement. PMID- 12723055 TI - Prediction of cyclosporine dosage in patients after kidney transplantation using neural networks. AB - This paper proposes the use of neural networks for individualizing the dosage of cyclosporine A (CyA) in patients who have undergone kidney transplantation. Since the dosing of CyA usually requires intensive therapeutic drug monitoring, the accurate prediction of CyA blood concentrations would decrease the monitoring frequency and, thus, improve clinical outcomes. Thirty-two patients and different factors were studied to obtain the models. Three kinds of networks (multilayer perceptron, finite impulse response (FIR) network, and Elman recurrent network) and the formation of neural-network ensembles are used in a scheme of two chained models where the blood concentration predicted by the first model constitutes an input to the dosage prediction model. This approach is designed to aid in the process of clinical decision making. The FIR network, yielding root-mean-square errors (RMSEs) of 52.80 ng/mL and mean errors (MEs) of 0.18 ng/mL in validation (10 patients) showed the best blood concentration predictions and a committee of trained networks improved the results (RMSE = 46.97 ng/mL, ME = 0.091 ng/mL). The Elman network was the selected model for dosage prediction (RMSE = 0.27 mg/Kg/d, ME = 0.07 mg/Kg/d). However, in both cases, no statistical differences on the accuracy of neural methods were found. The models' robustness is also analyzed by evaluating their performance when noise is introduced at input nodes, and it results in a helpful test for models' selection. We conclude that neural networks can be used to predict both dose and blood concentrations of cyclosporine in steady-state. This novel approach has produced accurate and validated models to be used as decision-aid tools. PMID- 12723056 TI - Detection of seizure precursors from depth-EEG using a sign periodogram transform. AB - Brief bursts of focal, low amplitude rhythmic activity have been observed on depth electroencephalogram (EEG) in the minutes before electrographic onset of seizures in human mesial temporal lobe epilepsy. We have found these periods to contain discrete, individualized synchronized activity in patient-specific frequency bands ranging from 20 to 40 Hz. We present a method for detecting and displaying these events using a periodogram of the sign-limited temporal derivative of the EEG signal, denoted joint sign periodogram event characterization transform (JSPECT). When applied to continuous 2-6 day depth-EEG recordings from ten patients with temporal lobe epilepsy, JSPECT demonstrated that these patient-specific EEG events reliably occurred 5-80 s prior to electrical onset of seizures in five patients with focal, unilateral seizure onsets. JSPECT did not reveal this type of activity prior to seizures in five other patients with bilateral, extratemporal or more diffuse seizure onsets on EEG. Patient-specific, localized rhythmic events may play an important role in seizure generation in temporal lobe epilepsy. The JSPECT method efficiently detects these events, and may be useful as part of an automated system for predicting electrical seizure onset in appropriate patients. PMID- 12723057 TI - A pseudojoint estimation of time delay and scale factor for M-wave analysis. AB - A pseudojoint estimation of time scale and time delay between an unknown deterministic transient type signal and a reference signal is proposed. The method is based on the separation between the estimations of the two dependent parameters. The time autocorrelation function (TACF) preserves the time scale and is invariant with respect to the time delay between the signals. The time scale factor can, thus, be estimated independently from time delay using the TACFs of the two signals. After estimating the time scale factor, the signal can be scaled by the estimated amount. The time delay is then estimated without bias due to the time scale factor. To obtain high resolution joint estimates, the time scale factor is estimated in the scale domain from the scale transforms of the TACFs of the two signals. The proposed method has low computational cost. Moreover, the results on synthetic signals show good performance of the method with respect to the Cramer-Rao Lower Bound and the joint Maximum Likelihood Estimation. A possible application of the technique to the analysis of electromyogram (EMG) signals detected during electrically elicited contractions is also presented. In a few representative cases, it is shown that the time scale estimate reveals myoelectric manifestations of muscle fatigue and is less affected by M-wave truncation than spectral EMG attributes. PMID- 12723058 TI - Harmonic cut and regularized centroid transform for localization of subcellular structures. AB - Two novel computational techniques, harmonic cut and regularized centroid transform, are developed for segmentation of cells and their corresponding substructures observed with an epi-fluorescence microscope. Harmonic cut detects small regions that correspond to small subcellular structures. These regions also affect the accuracy of the overall segmentation. They are detected, removed, and interpolated to ensure continuity within each region. We show that interpolation within each region (subcellular compartment) is equivalent to solving the Laplace equation on a multi-connected domain with irregular boundaries. The second technique, referred to as the regularized centroid transform, aims to separate touching compartments. This is achieved by adopting a quadratic model for the shape of the object and relaxing it for final segmentation. PMID- 12723059 TI - Is it feasible to reconstruct body segment 3-D position and orientation using accelerometric data? AB - The analysis of the mechanics of the musculo-skeletal system during the execution of a motor task requires the determination of the instantaneous position and orientation of the body segments involved in relation to an inertial system of reference. By using adequately assembled uniaxial accelerometric sensors, an easy to-manage measurement system can be obtained that estimates the three-dimensional position and orientation (P&O) of a body segment through an appropriate analytical model. However, the extent to which experimental errors, in particular accelerometers (ACs) assembly inaccuracies, affect such estimation has never been systematically investigated. This paper systematically analyzes the sensitivity of analytical models of body segment P&O reconstruction through a six-AC system and a nine-AC system to different sources of experimental error. We simulated and statistically assessed the performance of these models in the case of body segment motions typical of movements under muscular control. The results obtained indicated that the inaccuracy in the orientation of the individual AC's active axes and the offset error in the AC responses were the major sources of P&O estimation errors. In particular, no accurate estimation of position was possible with the analytical models analyzed. Under the motion conditions simulated in this study, no substantial advantages were found in using a nine-AC system rather than a six-AC system. Considering that the magnitudes of the simulated experimental errors were quite low (< or = 0.1 deg: AC's orientation; < or = 10( 4) m: uncertainty of the distance between two ACs; < or = 10(-2) ms(-2): random error; 0.5 x 10(-2) ms(-2): offset error), the results indicate that none of the two ACs systems analyzed is suitable for body segment P&O estimation in routine biomechanical applications. PMID- 12723060 TI - Electromagnetic radiation from ingested sources in the human intestine between 150 MHz and 1.2 GHz. AB - The conventional method of diagnosing disorders of the human gastro-intestinal (GI) tract is by sensors embedded in cannulae that are inserted through the anus, mouth, or nose. However, these cannulae cause significant patient discomfort and cannot be used in the small intestine. As a result, there is considerable ongoing work in developing wireless sensors that can be used in the small intestine. The radiation characteristics of sources in the GI tract cannot be readily calculated due to the complexity of the human body and its composite tissues, each with different electrical characteristics. In addition, the compact antennas used are electrically small, making them inefficient radiators. This paper presents radiation characteristics for sources in the GI tract that should allow for the optimum design of more efficient telemetry systems. The characteristics are determined using the finite-difference time-domain method with a realistic antenna model on an established fully segmented human body model. Radiation intensity outside the body was found to have a Gaussian-form relationship with frequency. Maximum radiation occurs between 450 and 900 MHz. The gut region was found generally to inhibit vertically polarized electric fields more than horizontally polarized fields. PMID- 12723061 TI - Hepatic radiofrequency ablation with internally cooled probes: effect of coolant temperature on lesion size. AB - Radiofrequency (RF) ablation is a minimally invasive method for treatment of primary and metastatic liver tumors. One of the currently commercially available devices employs an internally cooled 17-gauge needle probe. Within the probe, cool water is circulated during ablation, which cools tissue close to the probe resulting in larger lesions. We evaluated the effect of different cooling water temperatures on lesion size. We created a finite-element method model, simulated 12 min impedance-controlled ablation and determined temperature distribution for three water temperatures. Lesion diameters in the model were 33.8, 33.4, and 32.8 mm for water temperatures of 5 degrees C, 15 degrees C, and 25 degrees C, respectively. We solved a simplified model geometry analytically and present dependence of lesion diameter on cooling temperature. We further performed ex vivo experiments in fresh bovine liver. We created four lesions for each water temperature, with the same water temperatures as used in the finite-element method (FEM) model. Average lesion diameters were 28.3, 30, and 29.5 mm for water temperatures of 5 degrees C, 15 degrees C, and 25 degrees C, respectively. Water temperature did not have a significant effect on lesion size in the ex vivo experiments (p = 0.76), the FEM model, and the analytical solution. PMID- 12723062 TI - Geometry-based finite-element modeling of the electrical contact between a cultured neuron and a microelectrode. AB - The electrical contact between a substrate embedded microelectrode and a cultured neuron depends on the geometry of the neuron-electrode interface. Interpretation and improvement of these contacts requires proper modeling of all coupling mechanisms. In literature, it is common practice to model the neuron-electrode contact using lumped circuits in which large simplifications are made in the representation of the interface geometry. In this paper, the finite-element method is used to model the neuron-electrode interface, which permits numerical solutions for a variety of interface geometries. The simulation results offer detailed spatial and temporal information about the combined electrical behavior of extracellular volume, electrode-electrolyte interface and neuronal membrane. PMID- 12723063 TI - Microelectrode array on folding polyimide ribbon for epidural mapping of functional evoked potentials. AB - This paper describes a flexible polyimide microelectrode ribbon with a bending structure for epidural recording. Bending the ribbon establishes good electro mechanical contact of the electrodes to the cortex. Impedance spectroscopy and noise measurements in vitro were used to characterize the electrodes. Our in vivo experiments then successfully achieved synchronized neuronal activity and evoked potential patterns useful for understanding brain function and for finding cortical function. PMID- 12723064 TI - An improved reconstruction algorithm for 3-D diffraction tomography using spherical-wave sources. AB - Diffraction tomography (DT) is an inversion technique that reconstructs the refractive index distribution of a weakly scattering object. In this paper, a novel reconstruction algorithm for three-dimensional diffraction tomography employing spherical-wave sources is mathematically developed and numerically implemented. Our algorithm is numerically robust and is much more computationally efficient than the conventional filtered backpropagation algorithm. Our previously developed algorithm for DT using plane-wave sources is contained as a special case. PMID- 12723065 TI - A comparison of algorithms for detection of spikes in the electroencephalogram. AB - Identification of the short transient waveform, called a spike, in the cortical electroencephalogram (EEG) plays an important role during diagnosis of neurological disorders such as epilepsy. It has been suggested that artificial neural networks (ANN) can be employed for spike detection in the EEG, if suitable features are provided as input to an ANN. In this paper, we explore the performance of neural network-based classifiers using features selected by algorithms suggested by four previous investigators. Of these, three algorithms model the spike by mathematical parameters and use them as features for classification while the fourth algorithm uses raw EEG to train the classifier. The objective of this paper is to examine if there is any inherent advantage to any particular set of features, subject to the condition that the same data are used for all feature selection algorithms. Our results suggest that artificial neural networks trained with features selected using any one of the above three algorithms as well as raw EEG directly fed to the ANN will yield similar results. PMID- 12723066 TI - A simple system for detection of EEG artifacts in polysomnographic recordings. AB - We present an efficient parametric system for automatic detection of electroencephalogram (EEG) artifacts in polysomnographic recordings. For each of the selected types of artifacts, a relevant parameter was calculated for a given epoch. If any of these parameters exceeded a threshold, the epoch was marked as an artifact. Performance of the system, evaluated on 18 overnight polysomnographic recordings, revealed concordance with decisions of human experts close to the interexpert agreement and the repeatability of expert's decisions, assessed via a double-blind test. Complete software (Matlab source code) for the presented system is freely available from the Internet at http://brain.fuw.edu.pl/artifacts. PMID- 12723067 TI - In vitro measurement of myocardial impedivity anisotropy with a miniature rectangular tube. AB - Due to rapid change of fiber orientation, it is difficult to measure myocardial impedivity separately in a longitudinal or transverse fiber direction without mutual influence in the two directions. Previously published values of the longitudinal and the transverse myocardial impedivity were derived indirectly from measurements that mixed the impedivity in all directions. Those values are questionable because the derivations were based on a simplified uniform myocardial fiber model. In this paper, a miniature rectangular tube was devised to facilitate direct measurement of myocardial impedivity in a uniform fiber direction. The average transverse-to-longitudinal ratio of the measured in vitro swine myocardial impedivity was about 1.66 from 1 Hz to 1 kHz and dropped to 1.25 at 1 MHz. The result is important for accurate modeling of the electrical property of myocardium in biomedical research of radio-frequency cardiac catheter ablation. PMID- 12723068 TI - Influence of hormone supplementation to extended semen on artificial insemination, uterine contractions, establishment of a sperm reservoir, and fertility in swine. AB - This study was performed to quantify the effect of hormone addition to semen using a low-fertility model to evaluate its effectiveness and mode of action. At 24 h after the onset of estrus, all gilts received a single low-dose AI (0.5 x 10(9) sperm/80 mL) with no hormone (control, C), estrogens (E, 11.5 microg), PGF2alpha (PG, 5 mg of Lutalyse), or oxytocin (OT, 4 IU), which were then evaluated for semen backflow (n = 48), oviductal and uterine sperm numbers (n = 28), uterine contractions (n = 12), pregnancy rate (PR, n = 120), and number of fetuses (n = 67). In Exp. 1, backflow of semen from the uterus was collected for 8 h after AI, whereas PR and fetuses were assessed at d 25 to 30 after AI. In Exp. 2, backflow was collected and reproductive tracts flushed to determine sperm numbers in the oviducts and the anterior segments of the uterus. In Exp. 3, sows were monitored for uterine contractions for 1 h before AI and for 2 h after AI. In Exp. 1, there was a treatment x time interaction for fluid loss (P < 0.001), but by 8 h after AI, there was no difference in the total volume (70 +/- 1 mL) of semen lost between hormone treatments (85%) compared to controls (90%). There was also a treatment x time interaction (P < 0.05) for number of sperm lost in the backflow (2.1 +/- 0.1 x 10(8)), but by 8 h following AI, there was no effect on total sperm lost for the hormone treatments (38%) compared to C (54%). There was a trend (P = 0.10) for increased numbers of sperm in the uteri of hormone-treated gilts (6.0 +/- 1.3 x 10(4)) compared with C gilts (2.2 +/- 1.3 x 10(4), but there was no effect of treatment on sperm numbers in the oviducts (3.2 +/- 1.3 x 10(4)). Within 0.5 h of AI, there was an increase in the frequency of contractions for PG compared with the other treatments (14.2 vs. 6.3/h, P < 0.005), however there was no effect on amplitude (54 mmHg) or duration (35 s) of contractions. The PR was not influenced by treatment and averaged 54% (P > 0.60), but total numbers of healthy fetuses were increased (P < 0.04) by PG (8.7) and tended (P = 0.06) to be increased for OT (8.4), but not for E (7.2) compared to C (5.8). Hormone addition to semen increased numbers of fetuses and this may be related to an alteration in the pattern of fluid and sperm loss after AI and a tendency for increased numbers of sperm in the anterior segment of the uterus. Therefore, in situations of lowered fertility, hormone addition could be a strategy to limit infertility in swine. PMID- 12723069 TI - Synchronization of Bos indicus x Bos taurus cows for timed artificial insemination using gonadotropin-releasing hormone plus prostaglandin F2alpha in combination with melengestrol acetate. AB - Nonlactating Bos indicus x Bos taurus cows were used in three herds to determine the efficacy of different PGF2alpha treatments in combination with GnRH and melengestrol acetate (MGA) for a timed artificial insemination protocol. The start of the experiment was designated as d 0, at which time cows were assigned a body condition score and received 100 microg of GnRH. Cows were fed MGA (0.5 x mg x cow(-1) x d(-1)) on d 1 to 7. On d 7, cows received either a single injection of PGF2alpha (Lutalyse sterile solution; 25 mg; n = 297), a single injection of cloprostenol sodium (Estrumate; 500 microg; n = 297), or half the recommended dose of PGF2alpha (12.5 mg; n = 275) on d 7 and 8. On d 10, all cows were artificially inseminated and received 100 microg of GnRH. Pregnancy rates to the timed artificial insemination (39%) were not affected by treatment, herd, or treatment x herd. There was an effect (P < 0.01) of artificial insemination sire on timed artificial insemination pregnancy rate for one herd, but not the other two herds. Herd influenced (P < 0.05) 30-d pregnancy rates, but there were no treatment or treatment x herd effects as 72.3% of the cows became pregnant during the first 30 d of the breeding season. Results indicate that the type of PGF2alpha treatment administered 7 d after GnRH did not influence timed artificial insemination pregnancy rates in nonlactating Bos indicus x Bos taurus cows. PMID- 12723070 TI - Effect of restricted postweaning growth resulting from reduced floor and feeder trough space on pig growth performance to slaughter weight in a wean-to-finish production system. AB - The effect of reduced pig growth rate postweaning as a result of restricted floor space and feeder trough space on subsequent growth to slaughter was investigated in a wean-to-finish system. Crossbred pigs (n = 1,728) were used in a randomized block design with a 2 x 2 x 2 factorial arrangement of treatments: 1) floor space (high [0.630 m2/pig] vs. low floor space [0.315 m2/pig]), 2) feeder trough space (unrestricted [4 cm/pig] vs. restricted feeder trough space [2 cm/pig]), and 3) period of imposing floor- and feeder-trough-space treatments (12 vs. 14 wk postweaning). Growth performance was measured from weaning (5.5 +/- 0.01 kg of BW; 17 d of age) to slaughter (the end of wk 25 postweaning). From the end of the treatment period to the end of wk 25, pigs on all treatments had the same floor and feeder trough space. Pigs with low floor space had lower (P < 0.01) ADG, ADFI, and gain:feed ratio than those with high floor space, and were therefore lighter (P < 0.05) at the end of the postweaning treatment period. Pigs given the restricted feeder trough space had lower (P < 0.05) ADFI, similar (P > 0.05) ADG, and higher (P < 0.01) gain:feed ratio than those with unrestricted feeder trough space during the treatment period. Pigs in the 14-wk treatment period had higher (P < 0.01) ADG and ADFI, but lower gain:feed than those in the 12-wk treatment during that period. In the subsequent period, from the end of treatment to wk 25, there was an interaction (P < 0.05) between floor space and treatment period; the difference in ADG and gain:feed for pigs on low vs. high floor space was greater for the 14-wk than the 12-wk treatment period. However, low-floor-space pigs tended (P = 0.06) to be lighter than high-floor-space pigs at the end of wk 25 postweaning. Neither feeder trough space nor treatment period affected pig growth performance during the period from the end of treatment to wk 25. Carcass backfat and longissimus depths at the end of wk 25 were not influenced (P > 0.05) by treatment. In summary, pigs with restricted growth due to low floor space until either 12 or 14 wk postweaning had increased growth and feed efficiency in the subsequent period to wk 25 postweaning, with only a slight effect on BW and no effect on carcass measures. PMID- 12723072 TI - Effects of supplementary selenium source on the performance and blood measurements in beef cows and their calves. AB - On December 2, 1999, 120 pregnant cows were weighed, their body condition scored, and then sorted into six groups of 20 stratified by BCS, BW, breed, and age. Groups were assigned randomly to six, 5.1-ha dormant common bermudagrass (Cynodon dactylon [L.] Pers.) pastures for 2 yr to determine the effects of supplemental Se and its source on performance and blood measurements. During the winter, each group of cows had ad libitum access to bermudagrass/dallisgrass (Paspalum dilatatum Poir.) hay plus they were allowed limited access (1 to 4 d/wk) to a 2.4 ha winter-annual paddock planted in half the pasture. Treatments were assigned randomly to pastures (two pastures per treatment), and cows had ad libitum access to one of three free-choice minerals: 1) no supplemental Se, 2) 26 mg of supplemental Se from sodium selenite/kg, and 3) 26 mg of supplemental Se from seleno-yeast/kg (designed intake = 113 g/cow daily). Data were analyzed using a mixed model; year was the random effect and treatment was the fixed effect. Selenium supplementation or its source had no effect (P > or = 0.19) on cow BW, BCS, conception rate, postpartum interval, or hay DMI. Birth date, birth weight, BW, total BW gain, mortality, and ADG of calves were not affected (P > 0.20) by Se or its source. Whole blood Se concentrations and glutathione peroxidase (GSH Px) activity at the beginning of the trial did not differ (P > or = 0.17) between cows receiving no Se and cows supplemented with Se or between Se sources. At the beginning of the calving and breeding seasons, cows supplemented with Se had greater (P < 0.01) whole blood Se concentrations and GSH-Px activities than cows receiving no supplemental Se; cows fed selenoyeast had greater (P < or = 0.05) whole blood Se concentrations than cows fed sodium selenite, but GSH-Px did not differ (P > or = 0.60) between the two sources. At birth and on May 24 (near peak lactation), calves from cows supplemented with Se had greater (P < or = 0.06) whole blood Se concentrations than calves from cows fed no Se. At birth, calves from cows fed seleno-yeast had greater (P < or = 0.05) whole blood Se concentrations and GSH-Px activities than calves from cows fed sodium selenite. Although no differences were noted in cow and calf performance, significant increases were noted in whole blood Se concentrations and GSH-Px activities in calves at birth as a result of feeding of seleno-yeast compared to no Se or sodium selenite. PMID- 12723071 TI - Effect of source of energy and rate of growth on performance, carcass characteristics, ruminal fermentation, and serum glucose and insulin of early weaned steers. AB - Seventy-three crossbred steers (initial BW = 170.5 +/- 5.5 kg) from The Ohio State University (Exp. 1) and 216 crossbred steers (initial BW 135.4 +/- 4.4 kg) from the University of Illinois (Exp. 2) were used to determine the effect of source of energy and rate of growth on performance, carcass characteristics, and glucose and insulin profiles on early-weaned steers. Effects of the diets used in Exp. 1 and 2 on ruminal pH and VFA concentrations were quantified using ruminally fistulated steers (Exp. 3). Cattle were weaned at an average age of 119 d in all experiments and were allotted by age, BW, and breed to one of four diets: high concentrate, fed ad libitum (ALCONC), high-concentrate fed to achieve a gain of either 1.2 kg/d (1.2CONC) or 0.8 kg/d (0.8CONC), or high-fiber, fed ad libitum (ALFIBER). At 218 d of age, all steers were placed on the ALCONC diet until slaughter. Steers were implanted with Compudose at the initiation of all experiments and with Revalor-S when they were estimated to be 100 d from slaughter. When steers in Exp. 1 averaged 181 and 279 d of age, serum samples were collected to determine glucose and insulin concentrations. Steers were slaughtered when a fat thickness of 1.27 cm was reached (Exp. 1) or after 273 d on feed (Exp. 2). In Exp. 1, days in the feedlot (P < 0.01) and age at slaughter (P < 0.01) were lowest for ALCONC and ALFIBER steers, and greatest for 0.8CONC steers. Overall, ADG was greatest for ALCONC and lowest for 0.8CONC steers; feed efficiency was lowest (P < 0.01) for ALFIBER steers. Final BW did not differ (P > 0.57) among treatments. At 181 and 218 d of age, serum insulin was increased (P < 0.10) and intramuscular fat percentage was greatest (P < 0.07), respectively, for ALCONC steers. In Exp. 2, overall ADG (P < 0.06) and final BW (P < 0.04) were greatest for ALCONC and lowest for 1.2CONC and 0.8CONC steers. Overall feed efficiency was greatest for 0.8CONC and lowest for ALFIBER (P < 0.01). Growing phase diet did not affect marbling score at 218 d of age or at slaughter (P > 0.81). In Exp. 3, differences in ruminal pH after feeding may have been a consequence of increasing acetate (ALFIBER), propionate (ALCONC), or a combination of VFA (0.8CONC and 1.2CONC), respectively (diet x time after feeding, P < 0.10). Controlling growth by limit-feeding a high-concentrate diet for only 100 d does not extend the growth curve of early-weaned steers or enhance intramuscular fat deposition at slaughter compared to ad libitum intake of a high concentrate or high-fiber diet. PMID- 12723073 TI - Estimated copper requirements of angus and simmental heifers. AB - In Exp. 1, Simmental (n = 21) and Angus (n = 21) heifers, approximately 9 mo of age, were used in a 160-d study to determine the effect of dietary Cu on growth and Cu status. Two- or three-yr-old first-calf heifers (21 Angus and 21 Simmental) entering into their last trimester of pregnancy were used in Exp. 2 to estimate Cu requirements of the two breeds during gestation and early lactation. Treatments in both studies consisted of 0 (control), 7, or 14 mg of supplemental Cu (as CuSO4)/kg of DM. The control corn silage-based diets contained 6.4 and 4.4 mg of Cu/kg of DM in Exp. 1 and 2, respectively, and 1.2 mg of Mo/kg. Dietary Cu did not affect performance in either breed in Exp. 1. Copper supplementation generally did not affect plasma Cu concentrations in Angus heifers, but increased (P < 0.05) plasma Cu in Simmental heifers from d 37 until the end of Exp. 1. Final liver Cu concentrations were lower (P < 0.05) than initial concentrations in control Angus and Simmental heifers; however, liver Cu increased (P < 0.01) in Cu-supplemented heifers. In Exp. 2, Cu supplementation of the control diet increased (P < 0.05) plasma Cu during gestation and greatly increased (P < 0.01) liver Cu in both breeds. Calves born to cows not supplemented with Cu also had lower plasma Cu concentrations than Cu-supplemented calves by 73 d of age. In both studies, control Simmental heifers had lower (P < 0.05) plasma Cu concentrations than Angus on most sampling dates. When Cu was supplemented at 7 or 14 mg/kg of DM,few differences in plasma Cu concentrations were observed between breeds. Results suggest that Angus heifers have a lower minimal Cu requirement than Simmental. Based on liver Cu, the control diets containing 4.4 or 6.4 mg of Cu/kg of DM did not meet the Cu requirement of either breed during gestation and lactation or growth. Addition of 7 mg of Cu/kg of DM to the control diets met Cu requirements of both breeds. PMID- 12723074 TI - Effect of group size on performance of growing-finishing pigs. AB - Six hundred forty growing-finishing pigs (initial BW = 23.2 +/- 4.8 kg) were used in a 12-wk study (final BW = 95.5 +/- 10.2 kg) to quantify the effects of group size (10, 20, 40, and 80 pigs/pen) on performance, tail biting, and use of widely distributed feed resources. One single-space wet/dry feeder was provided for every 10 pigs, and floor allowance was 0.76 m2/pig in all treatment groups. Weight gain and feed intake were measured every 2 wk. At weighing, a tail-biting injury score was given to each pig. Blood samples were collected and analyzed for neutrophil:lymphocyte ratio before regrouping at the beginning of the experiment, 24 to 48 h after regrouping, and on the last day of each trial. The use of feeders by individual pigs was assessed by behavioral observations. Average daily gain for the entire 12-wk trial did not differ among group sizes (861, 873, 854, and 845 g/d for groups of 10, 20, 40, and 80, respectively; P > 0.10). During the first 2 wk, ADG was lower for pigs in groups of 40 (554 g/d) than pigs in groups of 10 (632 g/d; P < 0.05), but not pigs in groups of 20 or 80 (602 and 605 g/d, respectively). Average daily feed intake, feed efficiency, and variability in final BW within a pen also did not differ among group sizes. Tail-biting injury scores increased throughout the study, but did not differ among group sizes. Similar proportions of pigs were removed from the trial for health reasons, primarily due to tail biting, in all treatments. Individual pigs in each group size ate from most, if not all, of the feeders in the pen. There was no evidence of spatial subgrouping within the larger groups. The results suggest that housing growing-finishing pigs in groups of up to 80 pigs is not detrimental to productivity and health if space allowance is adequate and feed resources are evenly distributed. PMID- 12723075 TI - Effects of small ruminal boluses used for electronic identification of lambs on the growth and development of the reticulorumen. AB - Fifty-four male lambs were used to study the effects of two types of small electronic boluses on the dimensions and epithelial characteristics of their reticulorumen. Newborn lambs were assigned according to bolus type and age of application to the following treatments: 1) control (C, n = 21), without bolus; 2) mini (M, n = 21), identified with a 9.3 x 37.4-mm, 5.2-g bolus during the first week after birth; and 3) small (S, n = 12), identified with a 15.0 x 39.1 mm, 20-g bolus after weaning at wk 5, when lambs weighed more than 12 kg. After weaning, lambs were given ad libitum access to concentrate and barley straw. Six lambs were euthanized at the start of the experiment to measure initial reticulorumen characteristics. Ten lambs (five from M and five from C treatments) were slaughtered at weaning and 24 (eight per treatment) were slaughtered when they reached 24 kg. After bolus recovery, the reticulorumen was emptied and filled with polyurethane foam to obtain reticulorumen casts. Weight of the emptied reticulorumen and volume of the casts were measured. Four representative lambs from each treatment were also slaughtered at 24 kg, and their reticulorumen used to evaluate papillae size, number of dead cells, and degree of keratinization of both the reticulum wall and the rumen wall epithelia. Weight at weaning (13.8 kg), age at the end of fattening (65 d), and mortality rate (4%) did not differ among treatments. Retention rate for M and S boluses was 82.4 and 100%, respectively. Fresh weight and volume of the reticulorumen did not differ among treatments at weaning (130 g and 1,679 mL) or at the end of the fattening period (640 g and 5,931 mL). Lambs in the M treatment had greater (P < 0.05) rumen papillae size and lower (P < 0.10) keratinization than C lambs; values in the S lambs were intermediate between M and C lambs. Neither the M nor S type of bolus affected dimensions of the reticulorumen, but the earlier presence of M boluses induced a greater papillae size, with no negative effects on health and fattening performances of young lambs. PMID- 12723076 TI - Effects of feeding supplemental fat to beef cows on cold tolerance in newborn calves. AB - Our objectives were to examine the effects of added fat in late-gestation cow diets on neonatal response to cold. In Exp. 1, pregnant fall-calving heifers received control (n = 5), safflower seed (n = 5), or whole cottonseed (n = 5) diets. The hay-based, isonitrogenous, and isocaloric diets, fed for 47 d prepartum, contained 1.5, 4.0, and 5.0% fat for control, safflower, and whole cottonseed diets, respectively. At calving, calf BW and vigor score, as well as fat, lactose, and IgG in colostrum were not affected (P > 0.30) by diet. Heifers fed the safflower diet tended to have greater colostral solids (P < 0.10) than heifers fed the control or whole cottonseed diets. At 6.5 h of age, calves were placed in a 5 degrees C cold room for 90 min. Calf vigor, shivering, body temperature, and blood samples were taken every 15 min. During cold stress, calf body temperature decreased 0.7 degrees C (P < 0.03). Across all diets, shivering and serum glucose concentrations increased (P < 0.05), whereas calf vigor and cortisol concentrations decreased (P < 0.02) during cold exposure. In Exp. 2, pregnant spring-calving cows (n = 98) received a control (n = 47) or whole cottonseed (n = 51) supplement. Hay-based diets fed for 68 d prepartum contained 2.0 and 5.0% fat for control and whole cottonseed diets, respectively. Calf BW, vigor, shivering, dystocia score, time to stand, time to nurse, serum glucose concentrations, and serum IgG were not affected (P > 0.50) by diet. Between 30 and 180 min, body temperature of calves from dams fed the whole cottonseed supplement decreased (P < 0.05) more than calves from dams fed the control supplement. Serum glucose concentrations in calves were not affected by diet (P > 0.30). Serum cortisol concentrations tended (P < 0.09) to be greater for calves from dams fed whole cottonseed than control calves. When ambient temperature was < 6 degrees C, calves born to dams fed whole cottonseed had greater (P < 0.05) BW, tended (P < 0.1) to stand earlier, and had greater serum IgG concentrations. We conclude that calves from dams fed high-fat diets containing safflower or whole cottonseed respond similarly to cold stress, but these responses may not be consistent with greater cold resistance. In addition, high-fat dietary supplementation of late-gestation cows may only be beneficial during calving seasons with prolonged cold weather. PMID- 12723077 TI - Economic weights for feed intake in the growing pig derived from a growth model and an economic model. AB - Economic weights are obtained for feed intake using a growth model and an economic model. The underlying concept of the growth model is the linear plateau model. Parameters of this model are the marginal ratio (MR) of extra fat and extra protein deposition with increasing feed intake (FI) and the maximum protein deposition (Pd(max)). The optimum feed intake (FI0) is defined as the minimum feed intake that meets energy requirements for Pd(max). The effect of varying FI and MR on performance traits was determined. An increase in FI results in a larger increase in growth rate with lower MR. For a given MR, feed conversion ratio is lowest when FI equals FI0. Lean meat percentage (LMP) is largest for a low MR in combination with a low FI. The decrease in LMP with higher FI islargest when FI exceeds FI0. Economic weights for FI, MR and Pd(max) depend on FI in relation to FI0. Economic weights for FI are positive when FI is less than FI0 and negative when FI is larger than FI0. The MR has only then a negative economic weight, when FI is below FI0. Economic weights of FI and MR have a larger magnitude with lower MR and lower Fl. In contrast, economic weights for growth rate and FI derived from the economic model only change in magnitude and not in sign with different levels of these traits. The economic model always puts a negative economic weight on FI since it expresses profit due to a decrease in FI with constant growth rate and LMP. This holds the risk of continuous decrease in FI in pig breeding programs. In contrast, the use of growth models for genetic improvement allows direct selection for an optimum feed intake which maximizes feed efficiency in combination with maximum lean meat growth. It is concluded that recording procedures have to be adapted to collect the data necessary to implement growth models in practical pig breeding applications. PMID- 12723078 TI - Maternal grandsire, granddam, and sire breed effects on growth and carcass traits of crossbred cattle. AB - Postweaning growth, feed efficiency, and carcass traits were analyzed on 1,422 animals obtained by mating F1 cows to F1 (Belgian Blue x British breeds) or Charolais sires. Cows were obtained from mating Hereford, Angus, and MARC IIIHereford, 1/4 Angus, 1/4 Pinzgauer, and 1/4 Red Poll) dams to Hereford or Angus (British breeds), Tuli, Boran, Brahman, or Belgian Blue sires. Breed groups were fed in replicated pens and slaughtered serially in each of 2 yr. Postweaning average daily gain; live weight; hot carcass weight; fat depth; longissimus area; estimated kidney, pelvic, and heart fat (percentage); percentage Choice; marbling score; USDA yield grade; retail product yield (percentage); retail product weight; fat yield (percentage); fat weight; bone yield (percentage); and bone weight were analyzed in this population. Quadratic regressions of pen mean weight on days fed and of cumulative ME consumption on days fed were used to estimate gain, ME consumption and efficiency (Mcal of ME/kg of gain) over time (0 to 200 d on feed), and weight (300 to 550 kg) intervals. Maternal grandsire breed was significant (P < 0.01) for all traits. Maternal granddam breed (Hereford, Angus, or MARC III)was significant (P < 0.05) only for fat depth, USDA yield grade, retail product yield, fat yield, fat weight, and bone yield. Sire breed was significant (P < 0.05) for live weight, hot carcass weight, longissimus area, and bone weight. Sex class was a significant (P < 0.001) source of variation for all traits except for percentage Choice, marbling score, retail product yield, and fat yield. Interactions between maternal grandsire and sire breed were nonexistent. Sire and grandsire breed effects can be optimized by selection and use of appropriate crossbreeding systems. PMID- 12723079 TI - Evaluation of melatonin receptor 1a as a candidate gene influencing reproduction in an autumn-lambing sheep flock. AB - The effect of the melatonin receptor 1a (MTNR1A) gene on fertility and litter size in autumn lambing was studied with 373 ewes from a population of sheep selected for 10 yr for fertility in May and June matings. Animals were from a composite line of 50% Dorset, 25% Rambouillet, and 25% Finnsheep. Two restriction fragment length polymorphisms were present in the population, with allelic frequencies of 0.42 and 0.58 for an MnII polymorphism (alleles M and m, respectively) and of 0.34 and 0.66 for an RsaI polymorphism (alleles R and r, respectively). Genotypic frequencies for the polymorphisms were not independent, suggesting an association between them in foundation animals. Effects of MTNR1A genotype on fertility and litter size were evaluated using mixed linear model or REML procedures, but were not significant for matings involving ewes of all ages. However, in adult ewes (3 yr old and older), fertility of ewes of genotype mm was 10.0 (mixed model; P < 0.09) to 11.2% (REML; P = 0.03) less than that of other ewes. Genotypic effects associated with MTNR1A in adult ewes accounted for 23.8% of the estimated total additive genetic variance in fertility. Litter size was not significantly associated with MTNR1A genotype; adult ewes of genotype mm had approximately 0.11 fewer lambs per ewe lambing than ewes of other genotypes. Fertility in autumn lambing is lowly heritable, expressed only in females, and manifested relatively late in life and only in some management systems. Access to genetic markers would thus be advantageous in selection programs. PMID- 12723080 TI - Analyses of growth curves of nellore cattle by multiple-trait and random regression models. AB - The purpose of this study was to compare estimates of genetic parameters for sequential growth of beef cattle using two models and two data sets. Growth curves of Nellore cattle were analyzed using body weights measured at ages 1 (birth weight) to 733 d. Two data samples were created, one with 71,867 records sampled from all herds (MISS), and the other with 74,601 records sampled from herds with no missing traits (NMISS). Records preadjusted to a fixed age were analyzed by a multiple-trait model (MTM), which included the effects of contemporary group, age of dam class, additive direct, additive maternal, and maternal permanent environment. Analyses were by REML, with five traits at a time. The random regression model (RRM) included the effects of age of animal, contemporary group, age of dam class, additive direct, additive maternal, permanent environment, and maternal permanent environment. All effects were modeled as cubic Legendre polynomials. These analyses were also by REML. Shapes of estimates of variances by MTM were mostly similar for both data sets for all except late ages, where estimates for MISS were less regular, and for birth weight with MISS. Genetic correlations among ages for the direct and maternal effects were less smooth with MISS. Genetic correlations between direct and maternal effects were more negative for NMISS, where few sires were maternal grandsires. Parameter estimates with RRM were similar to MTM cept that estimates of variances showed more artifacts for MISS; the estimates of additive direct maternal correlations were more negative with both data sets and approached -1.0 for some ages with NMISS. When parameters of a growth model obtained by used for genetic evaluation, these parameters should be examined for consistency with parameters from MTM and prior information, and adjustments may be required to eliminate artifacts. PMID- 12723081 TI - Genetic evaluation of growth in nellore cattle by multiple-trait and random regression models. AB - The objective of this study was to identify issues in genetic evaluation of beef cattle for growth by a random regression model (RRM). Genetic evaluation data included 2,946,847 records of up to nine sequential weights of 812,393 Nellore cattle measured at ages ranging from birth to 733 d. Models considered were a five-trait multiple-trait model (MTM) and a cubic RRM. The MTM included the effects of contemporary group, age of dam class, additive direct, additive maternal, and maternal permanent environment. Both additive effects were assumed correlated. The RRM included the same effects as MTM, with the addition of permanent and random error effects. The purpose of the random error effect, which was in addition to a residual effect with constant variance, was to model heterogeneous residual variances. All effects in RRM were modeled as cubic Legendre polynomials. Expected progeny differences (EPD) were obtained iteratively using a preconditioned conjugate gradient algorithm. Numerically accurate solutions with RRM were not obtained until the random regressions were orthogonalized. Computing requirements of RRM were reduced by more than 50%, without affecting the accuracy by removing regressions corresponding to very low eigen-values and by replacing the random error effects with weights. Afterward, the correlations between EPD from RRM and from MTM for EPD on selected weights were between 0.84 and 0.89. For sires with at least 50 progeny, these correlations increased to 0.92 to 0.97. Low correlations were caused by differences in parameters. The RRM applied to growth i s prone to numerical problems. Estimates of EPD with RRM may be more accurate than those with MTM only if accurate parameters are applied. PMID- 12723082 TI - Evaluation models and genetic parameters for calving difficulty in beef cattle. AB - Calving difficulty was analyzed under threshold and linear models considering either a fixed or random herd-year effect. The aim of the study was to compare models for predicting breeding values according to the size of herd-year groups. When simulating data sets with small herds, in order to obtain an unbiased evaluation under a nonrandom and negative association of sire and herd effects, the best model for a practical evaluation was the fixed linear model. Field data included 246,576 records of the largest Charolais herds in France. Models were compared using the correlations of estimated breeding values between the different models. Although the best model from a theoretical point of view was a threshold model with a fixed herd-year effect, a linear model with a fixed herd year effect was the best choice from a practical point of view for predicting direct effects for calving difficulty in beef cattle and was a sufficient choice for predicting the associated maternal effects for data set with large herds. Correlations between direct estimated breeding values under the reference model and the fixed linear model and the random threshold model were 0.94 and 0.91, respectively. Correlations between the corresponding maternal estimated breeding values were 0.94 and 0.98. Heritabilities of direct effects were 0.27 and 0.14 under fixed threshold and fixed linear models, respectively. The corresponding heritabilities of maternal effects were 0.18 and 0.13, and the genetic correlation between direct and maternal effects were -0.36 and -0.34, respectively. PMID- 12723083 TI - Reallocation of body resources in lactating mice highly selected for litter size. AB - The present study investigated differences in the allocation patterns of body stores in lactating female mice from a line selected for high litter size at birth (S-line, average litter size of 20) and dams from a nonselected control line (C-line, average litter size of 10). Body weight, litter size, litter weight, and absolute and relative lipid and protein mass were measured at peak lactation (2 wk in lactation) and at weaning (3 wk in lactation). Body size in S line females has been increased as a correlated effect of selection for high litter size at birth, allowing for larger litters and higher absolute milk production. However, these dams produce larger litters relative to their own body weight. At peak lactation, lipid and protein percentage did not differ between lines. At weaning, S-line females had a higher protein percentage (P < 0.001) and lower lipid percentage (P < 0.05) than C-line females. Apparently, S-line females produce more offspring but at a greater cost to their own metabolism. This process was insufficient to supply the offspring with adequate resources, resulting in reduced (P < 0.0001) pup development and increased (P < 0.0001) preweaning mortality rates. PMID- 12723084 TI - A laser-based method to measure thermal nociception in dairy cows: short-term repeatability and effects of power output and skin condition. AB - To validate a laser-based method to measure thermal nociception in dairy cows (e.g., for the use in studies on stress-induced analgesia), we performed three experiments to observe the behavioral responses to a computer-controlled CO2 laser beam applied to the skin on the caudal aspect of the metatarsus. In Exp. 1, effects of power output (0, 1.3, 1.8, 2.2, 2.4 and 2.6 W) on nociceptive responses were examined using 18 dairy cows kept and tested in tie stalls. Increasing the power output affected the latencies to respond (decreasing latencies, P < or = 0.01), types of response (less nonresponding and more kicking, P < 0.0001), and behavior during (increasing frequency of tail flicking, P = 0.003) and between single laser exposures (increasing frequency of kicking, P = 0.02). Therefore, behavioral responses to a laser stimulus seem to be a valid measure of nociception in dairy cows. Repeatability within 15 min was investigated in Exp. 2 using n = 36 dairy cows kept and tested in tie stalls and a power output of 1.8 W. The variables' latency to move the exposed leg and frequency of tail flicking during laser exposure showed the highest level of repeatability (0.50 and 0.38, respectively). However, retesting at t = 15 min led to increased responses in terms of shorter latencies to respond (P < 0.05), increased kicking (P = 0.05), and tail flicking (P = 0.02), which probably can be explained by sensitization. Effects of power output (1.0 vs. 1.8 W) and skin condition (naked vs. intact) were examined in Exp. 3 on 11 group-housed dairy cows, tested just outside their home pen. Increasing the power output and shaving off hair led to increased responses as seen by shorter latencies to respond (P < 0.0001), less nonresponding (P < 0.0001), and increased kicking (P = 0.0003), as well as reduced intra- and interindividual variability (P < or = 0.04). In conclusion, the results of these experiments suggest that behavioral responses to laser stimulation are a valid and reliable measure of nociception in dairy cows, especially when applied on naked skin, both in the home environment and just outside a group pen. The fact that repeated testing in itself at t = 15 min led to increased responses means that the test will be a conservative measure of stress-induced analgesia. PMID- 12723085 TI - Fender design and insulation of farrowing huts: effects on performance of outdoor sows and piglets. AB - Two studies assessed sow and litter performance when design features of farrowing huts varied. A fender is a structure that extends out the front of the hut to create a veranda that prevents young piglets from leaving. In Exp. 1, 206 lactating sows and their litters were used to assess litter performance and the time required to process litters for two fender designs (short wooden [WS] vs. tall metal with a board front [MT-b]) and insulation status (insulated [IN] vs. uninsulated [UN]) farrowing huts. A significant fender x insulation interaction was observed for total litter weaning weight. Lighter litters weaned (P = 0.013) from WS fenders with UN huts compared with the other treatments. Fender design did not (P > 0.05) influence the time required to process a litter with one stockperson. Less total time (P = 0.001) was required to process a litter when two people were present (10.60 +/- 0.74 min) compared with one stockperson (14.52 +/- 0.74 min). There were no (P > 0.05) differences between IN and UN huts for temperature and relative humidity measurements. In Exp. 2, 331 lactating sows and their litters were used to determine sow and litter performance when using one of two fender front designs (boards [MT-b] vs. roller [MT-r]). The front design of the fender did not influence (P > 0.05) most litter performance measures. We conclude that fender design, fender fronts, and insulation effects did not have large influences on sow and litter performance in a West Texas environment. PMID- 12723086 TI - Growth factor messenger RNA levels in muscle and liver of steroid-implanted and nonimplanted steers. AB - Ribonuclease protection assays were used to measure steady-state semimembranosus muscle and/or hepatic levels of IGF-I, IGFBP-3, IGFBP-5, hepatocyte growth factor (HGF), and myostatin messenger RNA (mRNA) in steers implanted from 32 to 38 d with Revalor-S, a combined trenbolone acetate and estradiol implant. Insulin-like growth factor-ImRNA levels were 69% higher (P < 0.01, n = 7) in the livers of implanted steers than in the livers of nonimplanted steers. Similarly, IGF-I mRNA levels were 50% higher (P < 0.05, n = 7) in the semimembranosus muscles of implanted steers than in the same muscles from nonimplanted steers. Hepatic IGFBP 3 mRNA levels were 24% higher (P < 0.07, n = 7) in implanted steers than in nonimplanted steers. Hepatic HGF and IGFBP-5 mRNA levels did not differ between implanted and nonimplanted steers. Similarly, muscle IGFBP-3, IGFBP-5, HGF, and myostatin mRNA levels were not affected by treatment. Previous data from these same steers have shown that circulating IGF-I and IGFBP-3 concentrations were 30 to 40% higher (P < 0.01, n = 7) in implanted steers than in nonimplanted, control steers. Additionally, the number of actively proliferating satellite cells that could be isolated from the semimembranosus muscle was 45% higher (P < 0.01, n = 7) for implanted steers than for nonimplanted steers. Viewed together, these data suggest that increased muscle IGF-I levels stimulate increased satellite cell proliferation, resulting in the increased muscle growth observed in Revalor-S implanted steers. PMID- 12723087 TI - Slow fiber cluster pattern in pig longissimus thoracis muscle: implications for myogenesis. AB - Recent evidence implicates fiber type proportions as playing a role in meat eating quality, and in pigs it has been suggested that the slow oxidative fibers contribute to both juiciness and tenderness. The fiber distribution in pigs is different from that found in most other species, in which the various types of skeletal muscle fiber are distributed in a "checkerboard" pattern, because in pigs the slow oxidative fibers have a clustered distribution. The initial processes leading to fiber clustering are likely to occur during myogenesis, but the precise mechanistic aetiology of this patterning and whether the slow oxidative fiber clusters occur in a random or ordered fashion is unknown. In the present study longissimus thoracis muscle from Large White crossbred pigs was sampled at the 10th rib, 48 h postmortem. Transverse cryo-sections were cut and histochemically stained to allow the identification of the main muscle fiber types: slow oxidative, fast glycolytic, and fast oxidative glycolytic. Images of the sections were captured and analyzed using point processes and Voronoi Tesselations to examine the randomness and spatial distribution of the clusters of slow oxidative fibers found in pig longissimus thoracis muscle. The results showed that an assumption of complete spatial randomness can be rejected and that a mathematical model incorporating a minimum distance of 1.7 to 2.0 microm between cluster centers produced fiber patterns similar to those observed in the original transverse sections of the muscle. In addition, if it assumed that the central fiber in each cluster is derived from primary myoblast progenitors, these results suggest that there may be some degree of repulsion between the primary fibers during the initial stages of cluster formation. The mechanistic basis of such repulsion is not clear, but it is speculated that secreted factors, such as sonic hedgehog or myostatin may play a role. PMID- 12723088 TI - Effects of repetitive use of hormonal implants on beef carcass quality, tenderness, and consumer ratings of beef palatability. AB - Effects of repetitive use of anabolic implants on beef carcass quality, tenderness, and consumer ratings for palatability were investigated using crossbred steer calves (n = 550). Steers from five ranches were randomly allocated to one of 10 different lifetime implant strategies or to a nonimplanted control group. Cattle were implanted at some or all of five phases of production (branding, weaning, backgrounding, feedlot entry, or reimplant time). Carcasses from the control group had higher (P < 0.05) marbling scores than carcasses from steers in all other treatment groups. Implanting steers at branding, weaning, or backgrounding vs. not implanting steers at these production stages did not affect (P > 0.05) marbling scores. Steers implanted twice during their lifetime produced carcasses with higher (P < 0.05) marbling scores than did steers receiving a total of four or five implants. Steaks obtained from carcasses in the control group had lower (P < 0.05) shear force values and were rated by consumers as more desirable (P < 0.05) for tenderness like/dislike than steaks obtained from carcasses in all other treatment groups. Implanting steers at branding or weaning production stages did not affect (P > 0.05) steak shear force values, consumer ratings for like/dislike of steak tenderness, or percentage of consumers rating overall eating quality of steaks as satisfactory. Implanting steers at backgrounding vs. not implanting steers at this production stage increased (P < 0.05) steak shear force values, but did not influence (P > 0.05) consumer ratings for like/dislike of steak tenderness or percentage of consumers rating overall eating quality of steaks as satisfactory. Steaks from nonimplanted steers were rated as more desirable (P < 0.05) for overall eating quality than steaks from steers implanted two, three, four, or five times. Use of implants increased (P < 0.05) average daily gain by 11.8 to 20.5% from weaning to harvest compared with nonimplanted controls. Implant strategies increased (P < 0.05) hot carcass weight of steers by 8.9 to 13.8% compared with the control group. Use of implants also increased (P < 0.05) longissimus muscle area and decreased (P < 0.05) estimated percentages of kidney/pelvic/heart fat, but did not affect (P > 0.05) dressing percentage or adjusted fat thickness. Our findings suggest that beef quality, palatability, and production characteristics are influenced by lifetime implant protocols. PMID- 12723089 TI - Prerigor injection using glycolytic inhibitors in low-quality beef muscles. AB - The objective of this experiment was to determine the effect of prerigor injection of several glycolytic inhibitors on pH, color, tenderness, and related traits of low-value beef cuts. The semimembranosus, triceps brachii, and supraspinatus muscles from each of 10 steer carcasses were removed 1 h postmortem. Control samples remained in the carcass at 2 degrees C for 24 h. Prerigor muscles were injected and tumbled with 10% (by weight) of one of four solutions: sodium citrate (NaC; 200 mM), sodium fluoride (NaF; 200 mM), sodium acetate (NaA; 200 mM), and calcium chloride (CaCl2; 300 mM). All muscles treated with NaC and NaF showed the highest pH and glycogen content (P < 0.05), indicating that glycolysis was inhibited. Injection of NaC in semimembranosus and supraspinatus produced the tenderest meat (P < 0.05), showing a greater increase in tenderization at 3 d than at 7 d postmortem. Treatment did not affect color or oxidation-reduction potential, but all treated muscles tended to be more oxidative (higher oxidation-reduction potential). Sodium citrate was identified as a potential compound to enhance tenderness of prerigor muscle without altering color. Further studies are required to investigate its effect on palatability traits and to provide a basis for commercial application of the process. PMID- 12723090 TI - Growth performance, nutrient digestibility, and fecal microflora in weanling pigs fed live yeast. AB - Two experiments were conducted to evaluate the effects of live yeast supplementation on nursery pig performance, nutrient digestibility, and fecal microflora and to determine whether live yeast could replace antibiotics and growth-promoting concentrations of Zn and Cu in nursery pigs. In Exp. 1, 156 pigs were weaned at 17 d of age (BW = 5.9 kg) and allotted to a 2 x 2 factorial randomized complete block design (six or seven pigs per pen with six pens per treatment). Factors consisted of 1) dietary supplementation with oat products (oat flour and steam-rolled oats; 0 or 27.7%) and 2) yeast supplementation at 0 or 1.6 x 10(7) cfu of Saccharomyces cerevisiae SC47/g of feed. In Exp. 2, 96 pigs were weaned at 17 d of age and allotted to a 2 x 2 factorial randomized complete block design (four pigs per pen with six pens per treatment) with factors of 1) diet type (positive control containing growth-promoting concentrations of Zn, Cu, and antibiotics or negative control) and 2) live yeast supplementation (0 or 2.4 x 10(7) cfu of Saccharomyces cerevisiae SC47/g of feed). The inclusion of oat products in Exp. 1 decreased (P < 0.10) overall ADG and final BW. Yeast supplementation did not affect growth performance of pigs in Exp. 1 (P = 0.65); however, ADG in Exp. 2 was 10.6% greater (P < 0.01) and ADFI was increased by 9.4% (P < 0.10) in pigs supplemented with yeast in the positive control diet. Addition of Zn, Cu, and antibiotics to the diet improved gain:feed ratio during the prestarter period (P < 0.02) and overall (P = 0.10). In Exp. 1, inclusion of oat products increased (P < 0.01) total bacteria in feces when measured on d 10. Fecal lactobacilli measured on d 28 were reduced (P < 0.05) in pigs fed diets with oat products and yeast (interaction, P < 0.05). In Exp. 2, yeast supplementation decreased (P < 0.05) total bacteria and lactobacilli. Dietary yeast resulted in a greater (P < 0.05) yeast count in feces of pigs during the starter phase of Exp. 1. Yeast decreased (P < 0.10) the digestibility of DM, fat, and GE in the prestarter phase and DM, fat, P, and GE in the starter phase, whereas oat products increased the digestibility of DM, CP, fat, and GE (P < 0.05) in the prestarter phase. Results indicate that live yeast supplementation had a positive effect on nursery pig performance when diets contained growth promoting antimicrobials. Nonetheless, the response was variable, and the conditions under which a response might be expected need to be further defined. PMID- 12723091 TI - Effects of blood meal pH and irradiation on nursery pig performance. AB - A total of 720 nursery pigs in three experiments were used to evaluate the effects of blood meal with different pH (a result of predrying storage time) and irradiation of spray-dried blood meal in nursery pig diets. In Exp. 1, 240 barrows and gilts (17 +/- 2 d of age at weaning) were used to determine the effects of blood meal pH (7.4 to 5.9) in diets fed from d 10 to 31 postweaning (7.0 to 16.3 kg of BW). Different lots of dried blood meal were sampled to provide a range in pH. Overall (d 0 to 21), pigs fed diets containing blood meal had greater ADG (P < 0.05) and ADFI (P < 0.05) than pigs fed diets without blood meal. Ammonia concentrations in blood meal rose as pH decreased. However, blood meal pH did not influence (P > 0.16) ADG, ADFI, or gain:feed (G:F). In Exp. 2, 180 barrows (17 +/- 2 d of age at weaning) were used to determine the effects of post drying pH (7.6 to 5.9) and irradiation (gamma ray, 9.5 kGy) of blood meal on growth performance of nursery pigs from d 5 to 19 postweaning (6.8 to 10.1 kg of BW). One lot of whole blood was isolated with 25% of the total lot dried on d 0, 3, 8, and 12 after collection to create a range in pH. Overall, pigs fed blood meal had improved G:F (P < 0.01) compared to pigs fed the control diet. Similar to Exp. 1, the ammonia concentration of blood meal increased with decreasing pH. Blood meal pH did not influence ADG, ADFI, or G:F (P > 0.21), but pigs fed irradiated blood meal (pH 5.9) had greater ADG and G:F (P < 0.05) than pigs fed nonirradiated blood meal (pH 5.9). In Exp. 3, 300 barrows (17 +/- 6 d of age at weaning) were used to determine the effects of blood meal irradiation source (gamma ray vs. electron beam) and dosage (2.5 to 20.0 kGy) on growth performance of nursery pigs from d 4 to 18 postweaning (8.7 to 13.2 kg of BW). Overall, the mean of all pigs fed blood meal did not differ in ADG, ADFI, or G:F (P > 0.26) compared to pigs fed the control diet without blood meal. Pigs fed irradiated blood meal had a tendency (P < 0.10) for increased G:F compared with pigs fed nonirradiated blood meal. No differences in growth performance were detected between pigs fed blood meal irradiated by either gamma ray or electron beam sources (P > 0.26) or dosage levels (P > 0.11). These studies suggest that pH alone as an indicator of blood meal quality is not effective and irradiation of blood meal improved growth performance in nursery pigs. PMID- 12723093 TI - Content of ileal EAAC1 and hepatic GLT-1 high-affinity glutamate transporters is increased in growing vs. nongrowing lambs, paralleling increased tissue D- and L glutamate, plasma glutamine, and alanine concentrations. AB - Glutamate is a central metabolite for whole-animal energy and N metabolism. This study tested the hypothesis that ileal epithelium, liver, and kidney content of system X-(AG) glutamate transporters EAAC1 and GLT-1 would be up-regulated to support growth of wethers (30 +/- 1.2 kg) fed a forage-based diet for at least 14 d to gain (2.0 x NEm; n = 9) vs. maintain (1.2 x NEm; n = 9) BW. We have previously demonstrated that two high-affinity glutamate transporters (EAAC1, GLT 1) are expressed by these extensive glutamate metabolizing epithelial tissues. Wethers fed at 2.0 x NEm gained (P < 0.001; 0.26 kg/d) BW, whereas those fed 1.2 x NEm did not. Although plasma concentrations (microM) of glucose and L- or D glutamate did not differ, plasma glutamine (precursor of glutamate) and alanine concentrations (transamination product of glutamate) were 28% (P < 0.007) and 22% (P < 0.072) greater for growing lambs than nongrowing lambs. In tissues, the concentration of L-glutamate in ileum epithelia and D-glutamate of liver was 49% (P < 0.015) and 181% (P < 0.042) greater, respectively, in growing vs. nongrowing animals, whereas concentrations of glutamate isoforms did not differ in kidney. Paralleling these increased amino acid concentrations, ileal epithelium contained 313% more (P < 0.038) EAAC1 protein and liver contained 240% more (P < 0.001) GLT 1 protein, whereas kidney transporter content did not differ between growing and nongrowing wethers. In contrast to increased EAAC1 and GLT-1 protein content in ileal and liver tissue of growing lambs, messenger RNA levels did not differ. These results indicate that the increased capacity for high-affinity glutamate uptake in growing vs. nongrowing lambs is achieved through increased expression of EAAC1 by ileal epithelium and GLT1 by liver, which parallel increased tissue concentrations of glutamate and plasma concentrations of two major interorgan N carriers, glutamine and alanine. PMID- 12723092 TI - Season of the year influences testosterone secretion in bulls administered luteinizing honrmone. AB - The objective of the present study was to evaluate the secretion of testosterone (T) in bulls in response to the administration of varying doses of bovine LH (bLH) during the four seasons of the year. Five adult bulls (4 yr of age) were treated with an amount of bLH that was estimated to induce a 5 ng/mL amplitude pulse of LH in blood serum on five consecutive days around the spring equinox, summer solstice, fall equinox, and winter solstice. Five hours after this dose, bulls were treated with bLH in amounts that were estimated to induce a 0.25, 0.5, 1, 2, or 4 ng/mL amplitude LH pulse in blood serum in a Latin square design. Blood samples were collected for 5 h after administration of a dose of bLH that was estimated to induce the 5-ng amplitude LH pulse, and for 3 h after administration of the variable doses of bLH, and were then assayed for concentrations of T. Average concentrations and amplitude of T release after doses of bLH that were estimated to induce the 5-ng amplitude LH pulses were greater during the spring and summer than during the winter (P < 0.05). The area under the release curve (AUC) was greater during the spring than during the winter (P < 0.05). During the 3 h after treatment with the variable doses of bLH, T response was affected by dose (P < 0.001) and season (P < 0.001), but there was no dose x season interaction. Testosterone response increased in a dose-dependent fashion for all variables studied. The greatest average concentrations of T and AUC were observed in the spring compared with the fall and winter (P < 0.05). These data support our working hypothesis that testes of bulls are more responsive in releasing T in response to bLH stimulation in the spring and summer compared with the winter; however, there were no changes in sensitivity of the testes to LH during different seasons of the year as indicated by the lack of a dose of bLH x season interaction. PMID- 12723095 TI - Pancreatic exocrine secretion in steers infused postruminally with casein and cornstarch. AB - Our objective was to evaluate the effect of postruminal protein infusion on pancreatic exocrine secretions. One Holstein, two crossbred, and five Angus steers (305 +/- 5 kg) with pancreatic pouch-duodenal reentrant cannulas and abomasal infusion catheters were used in a replicated 4 x 4 Latin square. All steers were abomasally infused with 1,050 g/d of raw cornstarch with treatments of 0, 60, 120, or 180 g/d of sodium casein suspended in water to yield 6,000 g/d of infusate daily. Steers were limit-fed (1.5 x NEm; 12 equal portions daily) a 90% corn silage, 10% supplement diet formulated to contain 12.5% CP. Periods consisted of 3 d of adaptation to infusion, 7 d of full infusion, 1 d of collection, and 7 d of rest. Pancreatic juice was collected in 30-min fractions continuously for 6 h. Total juice secreted and the pH of individual fractions were recorded, a 10% subsample was retained to form a composite sample, and remaining fluid was returned to the duodenum. Juice composite samples were stored (-30 degrees C) until analyzed for total protein and activities of alpha-amylase, trypsin, and chymotrypsin. Casein infusion linearly increased alpha-amylase concentration (182 to 271 units/mL; P < 0.02; 17.5 to 24.6 units/mg of protein; P < 0.03) and secretion rate (26,847 to 41,894 units/h; P < 0.01). Total juice secretion (155 g/h), pH of pancreatic juice (8.13), secretion rate of protein (1,536 mg/h), and concentration of protein (10.2 mg/mL) in pancreatic secretions were not affected (P > 0.05) by casein infusion. Similarly, casein infusion did not change 0.05) trypsin and chymotrypsin concentrations (1,379 and 349 units/L or 0.134 and 0.033 units/mg of protein, respectively) or secretion rates (206 and 52 units/h, respectively). Abomasal infusion of protein with starch stimulated a greater pancreatic secretion of alpha-amylase activity into the intestine than infusion of starch alone. PMID- 12723094 TI - Influence of fibrolytic enzymes on the hydrolysis and fermentation of pure cellulose and xylan by mixed ruminal microorganisms in vitro. AB - A series of in vitro studies was conducted to determine the effects of adding a commercial enzyme product on the hydrolysis and fermentation of cellulose, xylan, and a mixture (1:1 wt/wt) of both. The enzyme product (Liquicell 2500, Specialty Enzymes and Biochemicals, Fresno, CA) was derived from Trichoderma reesei and contained mainly xylanase and cellulase activities. Addition of enzyme (0.5, 2.55 and 5.1 microL/g of DM) in the absence of ruminal fluid increased (P < 0.001) the release of reducing sugars from xylan and the mixture after 20 h of incubation at 20 degrees C. Incubations with ruminal fluid showed that enzyme (0.5 and 2.55 microL/g of DM) increased (P < 0.05) the initial (up to 6 h) xylanase, endoglucanase, and beta-D-glucosidase activities in the liquid fraction by an average of 85%. Xylanase and endoglucanase activities in the solid fraction also were increased (P < 0.05) by enzyme addition, indicating an increase in fibrolytic activity due to ruminal microbes. Gas production over 96 h of incubation was determined using a gas pressure measurement technique. Incremental levels of enzyme increased (P < 0.05) the rate of gas production of all substrates, suggesting that fermentation of cellulose and xylan was enzyme limited. However, adding the enzyme at levels higher than 2.55 microL/g of DM failed to further increase the rate of gas production, indicating that the maximal level of stimulation was already achieved at lower enzyme concentrations. It was concluded that enzymes enhanced the fermentation of cellulose and xylan by a combination of pre- and postincubation effects (i.e., an increase in the release of reducing sugars during the pretreatment phase and an increase in the hydrolytic activity of the liquid and solid fractions of the ruminal fluid), which was reflected in a higher rate of fermentation. PMID- 12723096 TI - Effect of grain processing and silage on microbial protein synthesis and nutrient digestibility in beef cattle fed barley-based diets. AB - Effects of the extent of grain processing and the percentage of silage in barley based feedlot diets on microbial protein synthesis and nutrient digestibility were evaluated using four steers (initial BW of 442 +/- 15 kg) with ruminal and duodenal cannulas. The experiment was a 4 x 4 Latin square with four periods of 21 d each. Dietary treatments were arranged as a 2 x 2 factorial with two levels of barley silage (20 and 5% DM basis) and two degrees of barley grain processing (coarsely and flatly steamrolled to a processing index [PI] of 86 and 61%, respectively). The PI was quantified as the volume weight of the barley grain after processing, expressed as a percentage of the volume weight prior to processing. Digest a flow (Yb) and microbial (15N) markers were continuously infused into the rumen for a period of 13 d. Ruminal, duodenal, and fecal samples were collected at various times over the last 6 d of marker infusion. Diurnal ruminal pH was measured for 48 h. Intake of DM averaged 1.8% of BW, and was not different among the dietary treatments (P > 0.10). Ruminal starch digestibility was higher (P < 0.05) for the more extensively processed grain and tended (P < 0.10) to be highest when the more extensively processed grain was combined with 5% barley silage. In contrast, ruminal fiber digestibility for the 5% silage diets was reduced (P < 0.05) when the grain was more extensively processed. There was, however, no effect of grain processing on ruminal OM digestibility (P > 0.10), and hence, no inhibitory effect on microbial N flow to the intestine (P > 0.10). There was also no effect of the level of silage on microbial N flow (P > 0.10), but there was a tendency for improved efficiency of microbial protein synthesis for the 20% silage diets (P = 0.072). Ruminal escape of nonmicrobial N (P = 0.003) was greater, and thus, protein flow to the intestine was greater for the 5% silage diets. Diurnal ruminal pH was lower (P < 0.05) for 11 of the 24 hourly time points in steers fed the 5% silage diets than those fed the 20% silage diets. In conclusion, barley grain rolled to a PI of 86 to 61% and combined with 20 and 5% barley silage had little effect on microbial protein supply. Microbial protein supply was not inhibited when the barley grain was extensively processed (PI of 61%) and the silage was limited to only 5% of the diet DM, but feed intake of steers in this study was lower than would be expected in the feedlot. PMID- 12723097 TI - Ryegrass-based diet and barley supplementation: partition of energy-yielding nutrients among splanchnic tissues and hind limbs in finishing lambs. AB - Splanchnic metabolism of energy-yielding nutrients and their uptake by the hind limb were studied in finishing lambs receiving ryegrass harvested at grazing stage (ear at 10 cm) with or without barley supplementation. Six ruminally cannulated and multicatherized lambs (40.2 +/- 1.5 kg) were fed with frozen ryegrass (RG) at 690 kJ of metabolizable energy intake (MEI) x d(-1) x BW(-0.75) successively with and without barley supplementation (RG + B), according to a triplicated Latin square design. Barley supplementation represented 21% of DM intake and increased the MEI by 32% (P < 0.002). In ruminal fluid, barley supplementation increased the acetate and butyrate concentrations by 21.2 and 49.6%, respectively (P < 0.04), without modifying those of propionate. Thus, molar proportions of acetate and butyrate were not modified, and those of propionate tended (P < 0.06) to decrease from 26 to 23%. As a result, the net portal appearance of propionate was not modified. Net portal appearance of butyrate and beta-hydroxybutyrate increased (P < 0.03), and that of acetate was not modified. Consequently, hepatic uptake of butyrate increased and probably spared acetate from hepatic metabolism. The hepatic fractional extraction of propionate decreased (P < 0.03), whereas the net flux of lactate switched from a net release to a net uptake, suggesting an alteration in the contribution of gluconeogenic substrates to glucose synthesis without modification in net hepatic glucose release. As a consequence, barley supplementation increased net splanchnic release of acetate (P < 0.02), propionate (P < 0.001), and beta hydroxybutyrate (P < 0.01) by 60, 157, and 78%, respectively. In addition, the net splanchnic release of insulin increased (P < 0.03) because of a decrease (P < 0.02) in its hepatic extraction. Despite those changes, the net uptake of nutrients by the hind limb was not modified and even decreased in the case of glucose (P < 0.02), suggesting a stimulation of lipogenesis in adipose tissues. Results from the present study suggested that supplementation of a ryegrass-based diet would likely have little effect on the orientation of muscle energy metabolism and on meat quality because the net uptake of nutrients by the hind limb was unchanged. PMID- 12723098 TI - Pre-harvest factors influencing the acid resistance of Escherichia coli and E. coli O157:H7. AB - The effects of pH, acetate, propionate, or butyrate concentration, and diet on acid resistance of fecal Escherichia coli and E. coli O157:H7 were determined by in vitro and in vivo experiments. The pH tested was from 4.0 to 8.0, and the VFA concentrations tested were 0 to 100 mM. The E. coli O157:H7 used was strain 505B. In an in vivo study, cattle were fed a grain-based diet, then either not switched or switched to a grain-based diet with 3% added calcium carbonate or two fiber based diets (soybean hulls or hay). Acid resistance was expressed as viability after acid-shock at pH 2.0 for 1 h and 4 h for fecal E. coli and E. coli O157:H7, respectively. Enumeration methods used were multitube fermentation, agar plate, and petri-film methods. The E. coli O157:H7 was not found in continuous culture inocula or in vivo samples. The viability of fecal E. coli decreased linearly (P < 0.01) as the culture pH increased, and viability of E. coli O157:H7 was highest (P < 0.01) when cultivated at pH 6.0. The viability of fecal E. coli and E. coli O157:H7 showed quadratic responses (P < 0.05) as acetate and butyrate concentrations increased at pH 7.2, with maximal acid resistance at 20 and 12 mM, respectively. As propionate concentration increased, the acid resistance was not different (P > 0.05) for fecal E. coli. Acid resistance of E. coli was induced by acetate and butyrate, even though the environmental pH was near neutral. Similar results were measured in the in vivo study, where viability after acid shock was more dependent on VFA concentration than on pH. Increasing the dietary calcium carbonate concentration also increased (P < 0.05) acid resistance of fecal E. coli. Results from these studies demonstrated that culture pH and VFA affect acid resistance of E. coli. PMID- 12723099 TI - Internationalization of the animal science undergraduate curriculum: a survey of its current status, barriers to its implementation and its value. AB - The goal of this project was to identify the current level at which internationalization has been adopted as a theme in the North American animal science curriculum and to identify its value and the barriers to its implementation. We surveyed animal, dairy, and poultry science departments across Canada and the United States. One hundred twenty-four surveys were mailed and 60% were returned. Associations between aspects of internationalization and student outcomes (admission to veterinary and graduate schools and starting salaries) were examined. Although administrators strongly believed internationalization had value, implementation was limited. The most common practices included international content in core animal science classes, advising, international internships, and participation of faculty in international scholarly activities. Few departments have incorporated internationalization into their mission statements or developed a specific international-themed class, scholarships devoted to international activities, or roles for international students. Few departments reported participation of students in international programs. Barriers included finances and limited commitment from higher administration. Student outcomes were positively associated with faculty size, percentage of international faculty, the ratio of international students to the total student population, international content in core animal science classes, a specific international-themed class, availability of international internships, and exchange of class material internationally via the Internet. Departments that did not offer international opportunities had a negative association (r = -0.79) with starting salary, but these relationships may not be causal. Alternatively, progressive departments may attract and retain exceptional students. The analysis indicated an awareness of the value of international programs, positive impacts in student outcomes, and financial barriers to implementation. PMID- 12723100 TI - An investigation into the effect of three months' clinical wear on toothbrush efficacy: results from two independent studies. AB - OBJECTIVE: Limited evidence has suggested that a worn toothbrush may be significantly less effective than a new brush with respect to plaque removal. Two independent studies, one with a manual toothbrush and one with a powered toothbrush, were undertaken to compared the ability of these toothbrushes as new and after three months' use to remove plaque. METHODOLOGY: Subjects from a general population were given either an Oral-B CrossAction manual toothbrush (Study 1) or a Braun Oral-B Battery (D4) toothbrush (Study 2). They were instructed to use their respective toothbrushes twice per day for a period of three months. After this time, those subjects meeting the studies' respective inclusion criteria entered the single-use, cross-over phase of the two studies. Both studies involved disclosing the teeth prior to measuring plaque. In Study 1, plaque was recorded using the Proximal Marginal Plaque Index and subjects brushed for one minute, by random assignment, with either a new or a worn toothbrush. In Study 2, plaque was measured using the Modified Quigley-Hein Plaque Index and subjects brushed for two minutes with either new or used brush heads, by random assignment. After a period of approximately one week, subjects returned to the test facilities and brushed with the alternate brush. RESULTS: Study 1 showed that both the new and the worn CrossAction toothbrushes significantly reduced plaque scores (p < 0.0001). A comparison of the two toothbrushes revealed no significant difference for whole-mouth plaque scores; however, at approximal sites the new toothbrush was significantly more effective than the worn brush (p = 0.033). In Study 2, as in Study 1, brushing with either a new or a worn brush head significantly reduced plaque scores (p < 0.0001). A comparison of the efficacies of the new and worn D4 toothbrushes revealed a non-significant tendency for the new brush head to remove more plaque than the worn brush head. However, when plaque removal was assessed for subjects using brush heads with the most extreme wear, i.e., scores of 3 or 4 (n = 15), a significant difference (p < 0.05) between new and worn brush heads was observed for the whole-mouth and approximal surfaces. CONCLUSION: The results from these two studies provide further data in support of the hypothesis that a worn toothbrush is less efficient with respect to plaque removal than a new brush. Patients should therefore be encouraged to replace their toothbrush regularly before bristle wear becomes excessive. PMID- 12723101 TI - Comparison of plaque removal efficacy of a battery-powered toothbrush and a manual toothbrush: a single-use clinical study in New Jersey. AB - OBJECTIVE: This single-use, examiner-blind clinical study evaluated the efficacy of a newly introduced battery-powered toothbrush (Colgate Motion Toothbrush) relative to a manual toothbrush (Oral-B CrossAction toothbrush) for the removal of supragingival plaque. METHODOLOGY: This study assessed plaque removal via the comparison of pre- and post-brushing plaque levels. A total of 126 adult male and female subjects from the northern New Jersey area reported to the clinical facility for a baseline (pre-brushing) plaque examination after having refrained from all oral hygiene procedures and chewing gum for 24 hours, and from eating, drinking or smoking for four hours. Subjects were entered into the study and stratified into two balanced groups based on their baseline plaque scores. Subjects were instructed to brush their teeth for one minute under supervision with their assigned toothbrush and a commercially available dentifrice, after which they were once again evaluated for supragingival plaque (post-brushing). RESULTS: All 126 subjects completed all aspects of the single-use clinical study. The subjects who used the Colgate Motion toothbrush exhibited a statistically significant 42.1% greater plaque reduction after a single tooth brushing than did those subjects who used the Oral-B CrossAction toothbrush. Relative to the pre brushing baseline scores, the Colgate Motion toothbrush group exhibited a statistically significant 59.0% reduction in plaque removal. CONCLUSION: The results of this single-use, examiner-blind, clinical study support the conclusion that the battery-powered Colgate Motion toothbrush provides significantly greater efficacy for the removal of supragingival plaque than does the manual Oral-B Cross-Action toothbrush. PMID- 12723103 TI - Comparative clinical study in plaque removal efficacy of a new sonic toothbrush (Float-Brush) with floating bristle action. AB - OBJECTIVE: Floating action toothbrush bristles have been incorporated into a V shaped sonic toothbrush. The base of the bristles moves down or up according to the pressure of the toothbrush being applied to a tooth surface. It was thought that this floating action with sonic vibration may generate new motion for increased plaque removal efficacy through increased contact between bristles and tooth surfaces. The objective of this investigation was to compare the plaque removal efficacy of a sonic toothbrush with floating bristle action (Float-Brush) to either a conventional V-shaped sonic toothbrush (Techno-Brush) or a manual toothbrush. MATERIALS AND METHODS: A single-blind, three-treatment cross-over study was performed on 42 subjects. Each subject refrained from brushing for 24 hours, followed by assessment for dental plaque on the Ramfjord teeth employing the Rustogi Modified Navy Plaque Index. Plaque removal efficacy was determined by the percentage of plaque score reduction in a single-use toothbrushing under supervision for two minutes. RESULTS: Forty participants completed this study. No significant difference in pre-brushing plaque scores was detected among the three toothbrushes. In the comparison of whole mouth mean scores, plaque removal efficacy of the Float-Brush (65.0%) was significantly higher than that of the Techno-Brush (59.1%, p = 0.0487) and the manual toothbrush (50.9%, p < 0.0001). Superiority of the Float-Brush was similar to whole mouth scores, when comparing the gumline and interproximal tooth area scores. CONCLUSION: These findings indicate the superiority of the Float-Brush for plaque removal compared to the Techno-Brush and the manual toothbrush. PMID- 12723102 TI - Comparative efficacy of two battery-powered toothbrushes on overnight plaque removal: a single-use clinical study in New Jersey. AB - OBJECTIVE: The objective of this single-use, examiner-blind clinical study, was to evaluate the efficacy of a newly designed Colgate Actibrush (battery-powered toothbrush) relative to the Crest SpinBrush (battery-powered toothbrush) for the removal of supragingival plaque. METHODOLOGY: This study included the assessment of plaque removal via the comparison of pre- and post-brushing plaque levels. A total of 80 adult male and female subjects from the central New Jersey area reported to the clinical facility for a baseline (pre-brushing) plaque examination after having refrained from all oral hygiene procedures and chewing gum for 24 hours, and from eating, drinking, or smoking for four hours. Subjects were entered into the study and stratified into two balanced groups based on their baseline plaque scores. Subjects were instructed to brush their teeth for one minute under supervision with their assigned toothbrush and a commercially available toothpaste (Colgate Cavity Protection Great Regular Flavor Fluoride Toothpaste), after which they were once again evaluated for supragingival plaque (post-brushing). RESULTS: All 80 subjects complied with the protocol, and completed the single-use clinical study. The subjects assigned to the Colgate Actibrush group exhibited a statistically significant 44.5% greater whole-mouth plaque reduction after a single brushing than did those subjects assigned to the Crest SpinBrush group. Relative to the pre-brushing baseline scores, the Colgate Actibrush group exhibited a statistically significant 63.7% reduction in whole mouth plaque removal. CONCLUSION: The results of this single-use, examiner-blind, clinical study support the conclusion that the newly designed Colgate Actibrush provides significantly greater efficacy for the removal of supragingival plaque than does the Crest SpinBrush. PMID- 12723105 TI - The Texas meeting teaser. PMID- 12723104 TI - Anticalculus efficacy of a chewing gum with polyphosphates in a twelve-week single-blind trial. AB - OBJECTIVE: A twelve-week clinical study was conducted to evaluate the effect of a chewing gum containing pyrophosphate (1%) and tripolyphosphate (1%) on controlling supragingival calculus deposits. METHODOLOGY: One-hundred and seventeen subjects enrolled in the study were randomly assigned to either a test gum or no gum group. Each participant received a full oral prophylaxis. Those chewing the test gum agreed to chew four times a day for five minutes each time over twelve weeks. Those assigned to the no gum group agreed not to chew any gum. After twelve weeks, participants were scored for calculus deposits using the modified Volpe-Manhold Calculus Index (VMI), received a second oral prophylaxis and entered the alternate group in this cross-over format. At the end of the twenty-fourth week, all subjects were again scored for calculus deposits and the study was completed. All participants received a supply of a sodium fluoride (0.32%) dentifrice for the study period. Calculus levels were scored by the same examiner for all measurements and recorded on computer. He was blinded to all treatment assignments. RESULTS: One-hundred and eleven participants completed the study. Six participants left the study but none reported problems linked to the pyrophosphates in the chewing gum. The VMI mean score for the test gum group was 2.55 (+/- 2.50) and the mean score for the no gum group was 4.09 (+/- 3.18). The difference between the mean scores was highly significant with paired sample t test (p < 0.0001). CONCLUSION: The results indicated that a chewing gum containing tripolyphosphate and pyrophosphate reduced calculus formation by 37.6% compared to a no gum treatment. PMID- 12723106 TI - Local anesthetics: special considerations in endodontics. AB - Local anesthetics are essential for successful endodontic treatment, and their pharmacologic characteristics have special implications for the treatment of painful, chronically-inflamed or necrotic teeth. Their dosages must be limited to prevent toxicity, which may be enhanced by the coadministration of sedative agents or drugs which affect hepatic drug metabolism. Endodontic patients with established central and peripheral sensitization represent special challenges for pain control, since morphogenetic changes resulting from neurogenic inflammation can render pain fibers more resistant to local anesthesia. The use of vasconstrictors with conventional and alternative injection techniques, e.g., intraosseous injections, are necessary to prolong the duration of action of local anesthesia but can place patients with cardiovascular disease at some risk. An appreciation of all of these aspects of local anesthesia in endodontics will better prepare the operator for predictably safe and effective patient care. PMID- 12723107 TI - Pulpalgia, the Pimpernel of pain. PMID- 12723109 TI - Use of nickel titanium instruments for cleaning and shaping root canal systems. AB - Rapid and significant changes in goals, techniques, instrument design, and type of metals used to manufacture endodontic instruments have been made over the last few years in an attempt to overcome canal preparation errors. Although techniques change, basic treatment objectives remain the same. Schilder has stated that "root canal systems must be cleaned and shaped: cleaned of their organic remnants and shaped to receive a three dimensional hermetic filing of the entire root canal space." The goal of root canal instrumentation is to obtain a continuously tapering funnel from the coronal access to the apex that flows with the shape of the original canal (1). The ability to prepare the canal without weakening the remaining dentin or perforating the root is essential for proper obturation and subsequent long-term success (2, 3). Procedural errors occurring during the instrumentation phase include apical zipping, instrument separation, canal transportation, ledging, and strip perforation (4-6). PMID- 12723108 TI - Strategies for managing the endodontic pain patient. AB - The most common form of orofacial pain is odontalgia, or toothache. This painful condition afflicts about 12-14 percent of the population, or about 2.5 million Texans (1-4). This is an important clinical challenge since our patients have high expectations for receiving the latest advances in health care, and successful pain control in endodontics is no exception. Fortunately, research has provided new approaches for delivering more effective pain control treatments in an efficient and reproducible manner. This article provides an overview of several treatment strategies for managing acute odontogenic pain; due to space limitations, interested readers are encouraged to use the references for more extensive discussions on these topics. PMID- 12723111 TI - Diagnosis and treatment planning: cracked tooth. AB - This article discusses the cracked tooth, one of the five major classifications of longitudinal tooth fractures: 1) craze line; 2) cuspal fracture; 3) cracked tooth; 4) split tooth; and 5) vertical root fracture. The term "longitudinal tooth fracture" was first introduced by Rivera (Personal Communication, Iowa City, IA, 1996) and has two meanings. The first implies distance (length), particularly in the vertical (occlusal-cervical) plane, as illustrated by longitudinal lines on a map. The second indicates that these fractures occur over a period of time. Therefore, the term longitudinal tooth fracture applies to fractures that have both a distance and a time component. Thus, fractures are described that are not related to impact trauma (which occurs primarily in incisors), in which the distance (length) component may be similar, but is immediate instead of over a period of time. PMID- 12723112 TI - Oral and maxillofacial pathology case of the month. Cicatricial pemphigoid. PMID- 12723114 TI - More bioterrorism. PMID- 12723115 TI - Terrorism: its impact on primary pediatrics, Part III. PMID- 12723110 TI - The management of traumatic dental injuries. AB - Traumatic dental injuries are classified typically as luxation, avulsion, and horizontal root fracture injuries. However, the most confusing aspect of treatment is generally centered on luxation and avulsion injuries. As clinicians, it can be somewhat confusing as to which regime is the most appropriate treatment, since there are a variety of guidelines and treatment modalities. PMID- 12723113 TI - What's driving up health insurance cost now? PMID- 12723116 TI - A 3-year-old girl with vomiting and a sixth nerve palsy. PMID- 12723117 TI - Industrial chemicals: terrorist weapons of opportunity. PMID- 12723118 TI - Cyanide as a weapon of terror. PMID- 12723119 TI - Diagnosis and management of nerve agent exposure. PMID- 12723120 TI - Vesicant agents and children. PMID- 12723121 TI - Weapons of mass destruction: the decontamination of children. PMID- 12723122 TI - Advocating for children during uncertain times. PMID- 12723123 TI - Graduating during a time of war. PMID- 12723124 TI - In her mother's voice: reflections on "femininity" and the superego. AB - For some female analysands the anxiety generated by superego conflict stimulated by aggressive and creative wishes may be warded off by invoking representations of the archaic all-powerful, limit-setting mother. To understand this clinical phenomenon in terms of a specific kind of female superego vulnerability is to miss the possibility that the fantasy of being mother's good little girl may be employed in the service of concealing the analysand's wishes and capacity for creative and destructive power. In the analytic hour we see transformed aspects of the analysand's past that are useful to the mind in its current operations. Representations of the mother and the archaic maternal imago are neither reflections of the past as it was, nor are they monolithic. These maternal representations may reverberate with adult memories of the past, but that remembered experience is a constructed one, constructed to serve particular mental operations. As these apparently remembered constructions arise in the current hour we need to remember that the present is the present, with its immediate desires and conflicts that the mind is attempting to represent. The past as it appears in the present is a new creation and, like the transference, a mutual creation of the analysand with her analyst. PMID- 12723127 TI - Thinking about thinking about "thinking about thinking". Neurobiology as the background for brain and mind. AB - This paper presents details of the development of recursive thought from early infancy to early childhood. During this period the child is said to face the task of developing a "representational theory of mind." In earlier writings I have suggested eight developmental stages of modeling of mind emerging from brain development. My emphasis has been that the selectionist neural Darwinian nature of the phylogenetic background for human brains and the accompanying nurture of the prolonged infant learning period for minds have together fashioned clear-cut steps of what the mind has to learn about thinking during this period. Object relations development of increasing sophistication emerges naturally from the recursive, back and forth communications between people and internally self reflective dialogues, which take place together. Both natural selection and social instruction based on that selection are seen as necessary aspects of this development. The contributions of Edelman, Tomasello, Barresi and Moore, and Fonagy and Target are each uniquely useful to catch these infant development concepts. My contributions of eight infant developmental stages by four-and-a half years of age fit with the findings of these others. PMID- 12723126 TI - Extra-transference interpretation. A defense of classical technique. AB - New psychoanalytic techniques stemming from various object relations and Kleinian frameworks have increasingly diverged from many of the principles of classical analysis, such as interpretations of genetic transference, resistances and reconstructions of the past. In contrast to the wide-ranging foci of classical technique, the so-called modern analysis is narrowly focused on the interpretation of transference in the here-and-now analytic relationship. This change in technique is attributed to paradigm shifts from Freud's intrapsychic metapsychology to two-person theories about internal object relationships, infantile phantasies and countertransference. This paper examines important elements of this broad issue through a specific focus on the controversy over the therapeutic value of extra-transference interpretations vs. here-and-now transference interpretations. An extensive review of the literature is followed by excerpts from published analyses to illustrate the different clinical styles of classical vs. modern analysis. Freud's actual analytic work with a patient is presented to show his natural, rule-free clinical style is very different from his early technique papers. Classical technique is also illustrated by a description of relevant aspects of the author's own training analysis with Anna Freud. Finally, a clinical vignette illustrates the author's approach to extra transference material and its interpretation. PMID- 12723125 TI - The history and current status of outcome research at the Anna Freud Centre. AB - In past decades there have been impressive systematic studies of the outcome of the psychoanalysis across groups of patients and analysts, but the studies carried out to date have almost all been with adult patients. There is very little information even on the short-term outcome of psychoanalytic treatment for children. The information that does exist has largely emerged from the research tradition of the Anna Freud Centre (formerly the Hampstead Clinic) in London. The paper surveys the early studies and details the current program of research at the AFC, providing a review of fifteen years of work on the outcomes of child psychoanalytic therapy. PMID- 12723128 TI - Dreams within dreams. AB - The concept of a dream within a dream is studied intensively. The illusion created by the dream work portrays one portion of a dream enacted within the envelope of another. Freud (1900) emphasized that the function of placing a piece of reality in a dream within a dream is an attempt to rob it of its significance and obliterate it. However, he seemed curiously disinterested in the fact that segmentation of a dream text into two seemingly discrete fragments does offer a dream investigator the opportunity to explore the dynamic relationship between the two fragments and the multiple meanings of the illusion created during sleep. In this study the linkage between the two parts of the dream sequence is highlighted. While Freud's intuition is corroborated, his lack of interest in the duality of the dream events is puzzling, as if he believed that only the dream within a dream is meaningful and the complementary and contextual dream sequence can be ignored. This paper suggests that both portions of the dream within a dream are significant, the one helping to explicate the other as the free associative process of dream interpretation gives equal democratic time to both. PMID- 12723129 TI - The superego. Its formation, structure, and functioning. AB - In this paper the theory of the superego is explored from the point of view of ego psychology. It traces the historical background in Freud's original contributions and the more contemporary understanding of the forces at work in the formation of this new psychic structure as they come together at a unique point in development, the oedipal phase. Superego functions are delineated, and precursors of superego functioning are differentiated from the functioning of the superego proper. Some attention is paid to the distinctions between ego and superego identifications, and between guilt and shame. Some clinical illustrations are included. PMID- 12723130 TI - Female sexuality. The pleasure of secrets and the secret of pleasure. AB - Why are secrets so fascinating and pleasurable, especially for little girls? A review of the psychoanalytic literature suggests that the ability to keep a secret represents an important developmental step. This paper proposes that secrets are integral features of female sexuality and that the sharing of secrets is important in how females relate to each other throughout development. It examines the role of sharing secrets in the psychoanalytic process and presents case material of female patients who cherished secrets and for whom the novel The Secret Garden was a favorite during childhood. PMID- 12723131 TI - The illusion of parental celibacy. A necessary stage in adolescent development. AB - The paper begins by reviewing Freud's case history of Dora and emphasizing her involvement in and overstimulation by her parents' sexual behavior. This markedly interfered with her ability to desexualize her relationship with them. As a result she was unable to develop the illusion of parental celibacy, which I postulate is an important and necessary defensive stage in normal adolescence. This illusion facilitates the desexualization of the adolescent's relationship to the parents and so contributes to separation from them and the seeking of non incestuous sexual outlets. The disruption of this illusion of parental celibacy by parental sex education, or by the complications of parental divorce may contribute significantly to the development of adolescent psychopathology. Clinical vignettes are presented. PMID- 12723132 TI - A psychoanalytical approach to integrating family and individual therapy in the treatment of adolescents. A case study. AB - An integrated model of object relations family therapy and a self psychology oriented individual therapy is presented in the form of a case study of an adolescent. The therapy strives to create and negotiate a balance between deconstructing the family's defensive delineations of the adolescent and helping him to form new building blocks of self-representation. The integrated model both highlights and adeptly addresses the unavoidable adolescent issue of conflicted loyalty between self and family as reflected in the adolescent's internal and external psychological world. PMID- 12723133 TI - Little orphan Anastasia, Part Two: developmental growth, love, and therapeutic change. AB - This is the second paper on a project that was undertaken to explore the opportunities and difficulties that might be encountered in writing about an ongoing child analytic case. For many reasons having to do with confidentiality and with the risks of reaching premature conclusions amid the normal confusions of the analytic process psychoanalysts generally write only after a case has been completed. Our project has sought to explore whether, as was the case with early psychoanalytic writings, the exercise of recording and synthesizing material gleaned from fresh open-ended psychoanalytic process might be accompanied by a more supple, informed, and nuanced clinical understanding. The first paper deals with two main questions. First, how could an infant survive and grow in the grim circumstances of a Siberian orphanage? Secondly, how do we understand the ways in which analytic technique interacted with Anastasia's extreme defensive activity to unstick her disorder of relatedness? Anastasia was born sickly. She was placed at two months in an orphanage where she developed a stool withholding disorder along with an attachment disorder. She was partially socialized in the home of the American parents who adopted her at nine months of age. However great gaps persisted between her intelligence and the use of her imagination and the torment and rage accompanying her infrequent bowel movements. Anastasia entered child analysis at age five and a half. The first paper described Anastasia's recovery from encopresis early in her child analysis and her subsequent struggle to overcome the psychological sequelae if her anti-dyadic state of relatedness. This paper will ponder some of the viscissitudes of transference and development that accompanied Anastasia's emerging capacity for love as she continued to overcome her alienation from her body and from significant others in her outside world. It will re-examine ways of thinking about object relationships and new objects to further understand Anastasia's recovery process. PMID- 12723134 TI - "The littlest balls ever company". The analysis of a five-year-old boy. AB - This paper describes the two-and-a-half year analysis of a five year old boy, Nick. Apart from the intrinsic interest that most child analytic cases hold, there are two especially noteworthy features of this analysis. First, Nick underwent an extensive psychological evaluation at the hands of the treating analyst some months before the treatment began. Thus, the case provides data about how that prior relationship affected the analytic process that evolved. Second, Nick initiated a project with the analyst that he named "the littlest balls ever factory." This project continued throughout the treatment, representing a variety of complex meanings. Although the genital and specifically testicular allusions in the name of this project seem obvious, the analyst came to understand that the project represented an equally important ego function: i.e., the enactment of an obsessional defensive process that allowed more dangerous and forbidden fantasies to emerge (A. Freud, 1936). This project represents a cautionary tale about not jumping to obvious conclusions. Nevertheless, the analyst was left with more questions than answers about the function of this all-important game to the patient. PMID- 12723136 TI - Escaping gravity. Movie magic and dreams of flying. AB - When movie-goers are thrilled by cinematic imagery portraying transcendent motor action, they are drawn into a world of imagination more often inhabited only in dreams of flying or of perfectly executed motion. The drive toward free and expansive movement which such dreams portray doesn't require explanation in terms of symbolic links to other aims, as has been common in the literature. The aim to achieve mastery in the locomotor sphere of action is a basic aim, in itself, integral to ego development. The developing child could not begin to organize perception, intention, and a coherent sense of self without this drive to move. This proposition is supported by emerging views of the ways the developing nervous system integrates systems mediating perception, action, and affect, which complement observations of the bodycentered development of mutual recognition in infants and their caregivers. PMID- 12723135 TI - Fathers facing their daughters' emerging sexuality. The return of the oedipal. AB - I propose that parents revisit their own oedipal passions and conflicts parallel to their children's passages through their oedipal phases. The child's oedipal behavior stirs up in the parent old attachment and separation issues as well as old desires and competitions. This both tests the parent's old solutions and offers the chance of new ones. In my view the parent not only is reacting to the child but is also experiencing the destabilization of his own intrapsychic structures. Such destabilizing, which occurs in any important period of psychological transition or development, is both promising and potentially dangerous; it opens up the possibility of more mature and more adaptive resolutions of revisited conflicts but also poses the danger of cracking a brittle or rigid adjustment, prompting a return to more primitive ways of coping. The resulting inner turmoil may produce disruptive thoughts, affects, and sometimes behavior, ranging from temporary benign regression to severe decompensation. It goes without saying that oedipal revisitations occur in both parents in conjunction with the oedipal stages of children of both sexes, but I focus here on paternal reactions to, and enactments with, daughters. It may be argued that not all men experience such a crisis, although my belief is that all fathers do. This paper illustrates the way it can appear in a number of different men. Awareness of this possibility enhances our understanding and clinical work with men and with their daughters. PMID- 12723137 TI - The character of the exception as a defense against deformity. AB - There is a tendency to regard Freud's paper on "the exception" (1916) as describing a general narcissistic defensive organization, but this paper argues that Freud was primarily concerned with the character of the exception as a defense against acknowledging bodily deformity. Two cases of bodily deformity, one fictional, one drawn from the author's practice, are presented to explore the role of magical identification with an ego-ideal in the regulation of self-esteem in the exception. The first shows a case irrevocably stuck in the character of the exception; the second example shows how analysis can help to effect some lessening of the defense. The paper ends with brief observations on technique in relation to the exception. PMID- 12723138 TI - The analysis of a pre-adolescent girl with primal scene fantasies. AB - This paper presents the analysis of Daphne, a ten-year-old girl whose prominent primal scene fantasies organized the analytic process in a rather unusual way, creating challenging technical problems and offering the analyst the opportunity to learn more about the formation of primal scene fantasies and their potential impact on psychic development and function. The data are consistent with the view that primal scene fantasies become organized in response to the cognitive requirements of development--to answer the enigmas of sexual difference, genital sensations, procreation, and parental rejection; that these fantasies arise from phase-specific libidinal and aggressive struggles such as voyeurism, exhibitionism, sadism, and homoerotic fantasy; and that they influence superego development. Daphne, however, appeared to have used the primal scene fantasy in a rather extraordinary way to organize and integrate the vicissitudes of development and especially to master conflict. Moreover, the analytic work demonstrates the technical difficulty of distinguishing between primal scene fantasy and primal scene exposure. Another technical issue was raised by the remarkable discrepancy between this little girl's relatively high functioning behavior and demeanor both in school and at home, and the outpouring of sexual material that emerged and flooded our sessions. Why was there such a discrepancy? How should I manage the sexualized nature of the material? Several colleagues, reflecting on the overstimulated quality of the analytic productions, questioned whether I "should have done something" to stop it. In the hours, I certainly felt perplexed by the discontinuity between her presentation and her fantasies. Thus, I ask the reader to follow me in the process of uncovering the particular way in which the analytic data revealed themselves. PMID- 12723139 TI - Language, transference, and the developmental context in child analysis. AB - The meaning of silence and its effect on the transferential relationship in child analysis are examined through the analysis of a ten-year-old boy. Silence is treated as a multiply determined symptom whose origins can be found in the early parent-child relationship but whose meaning is reworked and intertwined with wishes, both libidinal and aggressive in nature, that are aroused during later developmental phases. Silence is conceptualized as a powerful linguistic communication, for once sound is united with meaning, there can be no blank silence. Although silence also serves to defend against the expression of forbidden wishes, it cannot be treated purely as resistance; it is a defining boundary of the arena in which the patient has to be engaged in order for the analysis to proceed. The question of whether the patient needs to speak for the work of interpretation to be mutative is also addressed. PMID- 12723141 TI - The body in adolescent diaries. The case of Karen Horney. AB - The role of the body and its functions in the psychic life of the individual has occupied a central place in psychoanalytic thinking and writing. Developmentally, the body and the ego as a psychic organization have an integral, mutual relationship which becomes particularly important during adolescence, when the body matures physically while at the same time cognition, self-reflection, and social relations develop. This contribution presents results of the content analyses, focusing on the body, of 40 diaries written by twenty women during their adolescent years, compared with Karen Horney's adolescent diaries. In contrary to these diaries of the other young women, Karen Horney's adolescent diaries lack a focus on the body. Instead, idealized relationships with teachers are frequently mentioned. Only in the last diary, written at the transition to young adulthood, is the body, with a focus on sexual relations, more prominent. This discrepancy between Horney and female age-mates in normative samples is noteworthy. It is discussed here with emphasis on theoretical approaches about the body in adolescence in psychoanalysis, the importance of body and sexuality in Karen Horney's later life, Horney's contributions to female psychoanalysis, and her relationship with her father. PMID- 12723140 TI - The language of flowers. Freud's adolescent language of love, lust, and longing. AB - This paper traces the influence of Freud's adolescent language of flowers, a symbolic, metaphoric language of love, lust, and longing, from his youthful letters to Eduard Silberstein to later conceptualizations of his theory of sexuality, especially his 1905 theory of the transformations of puberty. In addition, information discovered by Boelich, editor of the Silberstein letters, sheds new light on the impetus for Freud's vigorous suppression of his adolescent poetic imagination and his adolescent crush on Gisela Fluss. The paper demonstrates that Freud's adolescent poetic imagination was transformed into a scientific imagination; the imagery and metaphors derived from the language of flowers were incorporated into his theory of masculinity, equated with aggressive sexuality, differentiated from femininity, characterized by submission and masochistic fantasies. By presenting details of Freud's experience from his autobiographic writings, the paper demonstrates his use of his personal struggle with adolescent fantasies in drawing generalizations about the role of fantasy in mental life. PMID- 12723142 TI - A visit to the Budapest school. AB - Within the past ten years, there has been a growing interest among American psychoanalysts in the work of Sandor Ferenczi and his followers, the so-called Budapest School. Several collections of essays on Ferenczi have appeared, while his correspondence with Freud and his Clinical Diary were finally published and translated. In this paper, I will give an overview of the historical importance of the Budapest School analysts, and then focus on three overlapping areas of their interest that have been and will continue to be particularly challenging. First, their exploration, stimulated by Ferenczi, of clairvoyance and telepathy, as these relate to the analytic situation, to transference and countertransference. Second, the mother-child dyad in the first year of life and the importance of trauma in early development. Third, the theory of psychosis. PMID- 12723143 TI - The influence of Marianismo on psychoanalytic work with Latinas. Transference and countertransference implications. AB - Marianismo, the traditional sexual code of behavior for Latinas, specifies chastity before marriage, sexual passivity after marriage, and the subordination of women to men. In addition to describing traditional gender role expectations for Latinos, this paper discusses marianismo as an important organizer of various conflicts experienced by Latinas. It also presents selected case material from a psychoanalysis to illustrate the impact of Marianismo on the therapeutic process when both the analyst and the patient are Latinas. PMID- 12723144 TI - A joke in a dream. A note on the complex aesthetics of disguise. AB - It was Freud's belief that while jokes and dreams have much in common, the comic elements that appear in dreams are not truly funny: unlike "real" jokes, they leave us cold. What, then, are we to make of the appearance in a dream of a well constructed joke that would produce laughter in and of itself even beyond the context of the dream? Such an example is cited and the reasons for the rarity of the phenomenon, as well as its multiple meanings, are explored. PMID- 12723145 TI - "Rage to order": Wallace Stevens and Hans Loewald. AB - In an essay published in 1988, "Psychoanalysis in Search of Nature: Thoughts on Metapsychology, Metaphysics, Projection," Hans Loewald animates both psychoanalysis and nature in their dynamic relation, one to the other. He recapitulates many of his earlier ideas about how the mind of the child "comes into being" in the resonating, transindividual field of the mother-infant dyad and applies them to how the individual person and nature form their transindividual field. The present paper uses Loewald's earlier ideas (1951 & 1960) and their elaboration in his late (1988) work to illuminate Wallace Stevens' poem "The Idea of Order at Key West." PMID- 12723146 TI - Distribution of the lanthanides in the earth's crust. PMID- 12723147 TI - Mobilization of lanthanides through the terrestrial biosphere. PMID- 12723149 TI - Biologically relevant structural coordination chemistry of simple lanthanide ion complexes. PMID- 12723148 TI - Complexes of lanthanide ions with amino acids, nucleotides, and other ligands of biological interest in solution. PMID- 12723150 TI - Lanthanide ions as probes in studies of metal ion-dependent enzymes. PMID- 12723151 TI - Lanthanide chelates as fluorescence labels for diagnostics and biotechnology. PMID- 12723152 TI - Responsive luminescent lanthanide complexes. PMID- 12723153 TI - Lanthanide ions as probes of electron transfer in proteins. PMID- 12723154 TI - Lanthanide ions as luminescent probes of proteins and nucleic acids. PMID- 12723155 TI - Lanthanide-promoted peptide bond hydrolysis. PMID- 12723156 TI - Lanthanide-catalysed hydrolysis of phosphate esters and nucleic acids. PMID- 12723157 TI - Sequence-selective scission of DNA and RNA by lanthanide ions and their complexes. PMID- 12723158 TI - Lanthanide ions as probes for metal ions in the structure and catalytic mechanism of ribozymes. PMID- 12723159 TI - Lanthanides as shift and relaxation agents in elucidating the structure of proteins and nucleic acids. PMID- 12723160 TI - Lanthanide ions as magnetic resonance imaging agents. Nuclear and electronic relaxation properties. Applications. PMID- 12723161 TI - Interactions of lanthanides and their complexes with proteins. Conclusions regarding magnetic resonance imaging. PMID- 12723162 TI - Metabolism and toxicity of the lanthanides. PMID- 12723163 TI - Cell responses to lanthanides and potential pharmacological actions of lanthanides. PMID- 12723164 TI - The effect of health care working conditions on patient safety. PMID- 12723165 TI - [Test your decision making skills. When should endodontic treatment be redone?]. AB - Research has shown wide variations in dentists' clinical attitude towards root canal treated teeth. Simulated cases are used as tools in the investigation of practitioners' clinical behavior and decision making. We therefore used simulated cases (clinical histories and radiographs) of 14 endodontically treated teeth to: (1) determine the predominant proposed treatment strategy for each case, (2) evaluate the consensus, if any, among practitioners, (3) identify pathology and dentist factors influencing the treatment choice. PMID- 12723166 TI - [Local anesthesia with and without vasoconstrictors: indications, contra indications and complications in relation to the health of the patient]. AB - Patient's fear of pain is one of the major reasons of not routinely seeking dental care. Local anesthetics are commonly used in dentistry and dental practitioners must be familiar with the effects and side-effects of these substances in medically compromised patients. This article is a review of these side-effects and the resulting contraindications. A frequently asked question is when to combine a vasoconstrictor to a local anesthetics. The purpose would be to reduce the anxiety and avoid any pain during dental procedures. Indeed, pain produces much more endogenous epinephrine than the small amount of epinephrine combined to the local anesthetics. PMID- 12723167 TI - [Electronic determination of the length of the root canal]. AB - The endpoint of an endodontic treatment must be situated at the cemen-to-dentinal junction to preserve the biological potential of the periapical area and to realize a complete disinfection of the principal root canal system. Thus, the determination of working length is a crucial factor in endodontic therapy. The conventional radiographic method cannot provide the accurate location of the apical constriction. For this reason, the electronic apex locators have been developed. Now, the last generation of apex locators has become more sophisticated and the electronic measurements seem to be accurate and reliable with electrolytic solutions in the canal. PMID- 12723169 TI - [Complications of refraction disorders]. AB - Refraction disorders, myopia, hypermetropy, astigmatism and anisometropy represent an important segment in ocular pathology as well as an economic and social problem for. Refractions disorders are also responsible for a series of complications generated by different levels of damages of the visual function. The purpose of this paper is to methodically present the possible complications at different types of refraction disorders at different ages. PMID- 12723168 TI - [Pitfalls to avoid in the cleaning and shaping of the pulp canal]. AB - Chemomechanical instrumentation of the root canal system is without doubt the most important but most challenging part of endodontic treatment. The first essential in cleaning and shaping is gaining direct access to the root canals. A properly prepared endodontic access opening can eliminate many of the technical problems encountered in root canal treatment. Excessive calcification in the root canal system and performing root canal treatment through existing artificial crowns however can be a challenge for the clinician. Secondly, enlargement of a curved root canal often results in loss of working length, incomplete removal or extrusion of the root canal content and unwanted alterations in the canal shape, such as transportation (stripping, zipping, elbow formation), ledge formation and perforation. This article focuses on the causes and effects of procedural errors during root canal cleaning and shaping. Each of these complications is addressed in a manner designed to prevent problems rather than to manage them once they are recognized. Throughout the immense amount of literature published the last 30 years on this topic, concensus is obtained concerning the preferential procedure of cleaning and shaping the root canal. PMID- 12723170 TI - [Postoperative endophthalmitis]. AB - PURPOSE: Is to establish the risk factors, the treatment and the prognostic factors at the patients operated for cataract, who developed earl endophtalmitis after the operation. MATERIAL AND METHOD: We studied 14 eyes which were operated for cataract who developed endophtalmitis after the the operation. RESULTS: At all patients the general treatment associated antibiotics (Cefalosporine) with steroids (high doses) and the local one was with mydriatics and steroids. After the treatment, we obtained a visual acuity between 0.7-1 at 11 eyes, visual acuity between 0.400,6 in 3 eyes and 2 eye had a visual acuity under 0.1. CONCLUSIONS: The postoperative endophthalmities is a complication which affect the functional prognosis of the patients operated for cataract. PMID- 12723172 TI - [Blue-yellow full threshold automated perimetry in glaucoma diagnosis]. AB - PURPOSE: To compare White & White and Blue-Yellow Full Threshold automated perimetry. MATERIAL AND METHOD: Group of 37 patients; we used a Humphrey Zeiss automated perimeter. RESULTS: We obtained 49% abnormal white & white visual fields and 62% abnormal blue yellow fields. CONCLUSIONS: Blue Yellow perimetry has higher sensibility than white & white perimetry. We can detect earlier with this test the glaucoma defects. PMID- 12723171 TI - [Secondary implantation of posterior chamber intraocular lenses]. AB - PURPOSE: Is to establish the possibilities of secondary implantation of posterior chamber intraocular lenses, complications and functional results obtained. MATERIAL AND METHOD: We revied 26 aphakic eyes which were secondary implanted with posterior chamber intraocular lenses s PC IoL). RESULTS: At 16 eyes were implanted standard PC IaL and at 1 o eyes we used scleral fixation PC IoL. Early postoperative complications of standard PC IoL were: uveitis in 4 eyes, dislocation at one case and late postoperative complications were: bulous keratopathy in one eye, secondary cataract in 4 eyes and at 3 eyes dislocations. At scleral fixation PC IQL we had in the early postoperative period secondary glaucoma in 2 eyes and in the late period one eye had retinal detchement and in 2 eyes persist ocular hypertonia. CONCLUSIONS: Posterior chamber intraocular lenses is the optimal solution in correction of aphakia. PMID- 12723173 TI - [Cytomegalovirus retinitis in HIV positive patients]. AB - First it is shown an up-date about cytomegalovirus retinitis, concerning pathogeny, diagnosis, evolution and treatment. Then is a study about incidence and evolution of cytomegalovirus retinitis in AIDS patients, which have the diagnosis among january 1994-september 2002. CONCLUSIONS: In Romania the incidence of cytomegalovirus retinitis in AIDS patients is lower than in western countries, because over 60% of patients are children. PMID- 12723174 TI - New concepts in etiology, clinic and management of postoperative endophthalmitis. AB - THE AIM OF THE PAPER: In the last few years, as a result of the continuing changing of the ethiological, clinical and therapeutical aspects of the postoperative endophthalmitis, there are a lot of efforts to find the adequate means to prevent and to treat this severe complication of cataract surgery. So, the paper presents the most actual therapeutical approaches in the field. MATERIAL AND METHODS: The goals of treatment of postoperative endophthalmitis are: to regain vision and to save the bulb. From the prophylactic techniques, preoperative povidone-iodine antisepsis is the most recommended procedure for decreasing the risk of endophthalmitis related to surgery. The main elements of the modern treatment of endophthalmitis are medical treatment by intravitreal injections or by systemic drugs and surgical treatment with vitrectomy. CONCLUSIONS: Although endophthalmitis remains a potential cause of visual loss, both medical therapies and surgical techniques have evolved so that prognosis is not always as negative as it used to be. PMID- 12723175 TI - [Trigeminal neuropathy, diagnostic and treatment problems]. AB - AIM OF THE STUDY: Clinical evaluation of patients with the diagnosis of Sturge Weber syndrome (encephalotrigeminal angiomathosis) for establishing an accurate diagnosis and therapy. MATERIAL AND METHOD: The paper presents 5 cases of encephalotrigeminal angiomathosis (4 of them associating glaucoma) admitted in our clinic in the last 5 years, pointing out the peculiarities concerning the clinical, diagnostic, therapeutically aspects and outcome. RESULTS: The analysis of our cases showed that glaucoma does not correlate with the extension of skin lesion (nevus flammeus). The outcome of glaucoma's therapy is dictated by the adjustment of the therapy to the type of glaucoma. CONCLUSION: Glaucoma associated with the Sturge-Weber syndrome has always proved to be a therapeutic challenge and remains one of the causes of increased intraocular pressure, difficult to control. The management of glaucoma in Sturge-Weber syndrome includes surgical and drug therapy. These patients need long-term follow-up from the clinical and imagistic point of vue. PMID- 12723176 TI - [Complications after catheterization of nasolacrimal duct]. AB - THE AIM OF THIS STUDY: Is to identify two severe complications after catheterism of nasolacrimal duct. A boy, 9 months old, was treated in Clinica I Oftalmologie lasi for presptal cellulitis after catheterism of nasolacrimal duct which was performed in other Eye clinic, without general anesthesia. Also, this child presented the rupture of punctal and inferior canalicular duct. CONCLUSION: Catheterism of nasolacrimal duct must be performed by ophthalmopediatric doctors with experience, under general anesthesia, to prevent complications. PMID- 12723177 TI - [Diabetes mellitus and glaucoma]. AB - PURPOSE: The evaluation of the incidence of the open angle glaucoma to the patients with diabetes mellitus compared to the incidence in the general population and the frequency of the neovascular glaucoma. MATERIAL AND METHOD: The work consists in a study of 1700 patients with diabetes mellitus who where examined during 13 years. RESULTS: From 1700 patients with diabetes mellitus, 240 (14.1%) have glaucoma (primary or secondary). Of those 240, 15 (6.2%) have diabetes mellitus type I and 225 (93.7%) have diabetes mellitus type II. Concerning the clinical form of glaucoma, 177 patients (10.4%) have primary glaucoma with open angle, 23 (1.3%) have primary glaucoma with narrow-angle and 40 (2.5%) have secondary glaucoma. CONCLUSIONS: The highly incidence of the primary glaucoma to the patients with diabetes mellitus comparative to the general population can be explained by the vegetative dysfunction in association with diabetes mellitus and the presence of some common immunitary anomalies. The small numbers of cases with neovascular glaucoma means an indicator of the efficiency of the antidiabetic treatment and of the ocular complications of the diabetes mellitus. PMID- 12723178 TI - [Phacoemulsification and diabetic eye]. AB - THE PURPOSE OF THIS STUDY: Is to display the influence of diabetes on the course of the operation, postoperative complications and functionals results in operated primary cataract. MATERIAL AND METHODS: We performed a study on 134 eyes with cataract and diabetes operated in the Universitary Eye Hospital and Optilens Eye Clinic from Cluj-Napoca. RESULTS: The majority of patients (83.58%) had noninsulinodependent diabetes. Background diabetic retinopathy was present in 27 cases, associated with the diabetic maculopathy in 8 eyes. In 3 cases was present preproliferative diabetic retinopathy and in one eye it was discovered proliferative retinopathy. EEC + PC-IOL was performed in 81 eyes, mini-nuc in 37 cases and in 16 eyes phacoemulsification with PC-IOL. Postoperative the complications were less in the cases operated with mini-nuc and phacoemulsification. Visual acuity was better in eyes without diabetic retinopathy. CONCLUSIONS: 1. The modern surgical techniques of cataract surgery reduce significant the frequency of postoperative complications. 2. Functional results depends on the presence of diabetic retinopathy. PMID- 12723179 TI - [Epidemiology of refraction disorders]. AB - THE PURPOSE OF THIS STUDY: Is to appreciate the prevalence and incidence of the refraction disturb among the children aged 7-11 years old. MATERIAL AND METHOD: Using the descriptive study method, we examined 3562 children aged between 7 to 11 years old, a representative group for Bihor Country. RESULTS: Among the studied children batch, hypermetropy represents 89.8%, myopia 1.4%, astigmatism 4.9% and anisotropy 4%. The amblyopy was diagnosed in 474 cases (13.3%). The medium amblyopy is predominant. CONCLUSIONS: The refraction disturb incidence is 9.5@1000 and their prevalence is 82.4@1000. In the studied group, the hypermetropy is predominant. In the group of children with 10 years old, we encountered the greatest number of myopia and emetropy. Amblyopic represents 13.3% from all cases. It is required an ophthalmological health program that will include the refraction disturb with children, as well. PMID- 12723180 TI - [Perforanting ocular trauma--comparative study]. AB - THE PURPOSE: Is the comparative study of surgically solved ocular trauma, within TARGOVISTE and BRASOV county Hospitals. MATERIAL AND METHOD: A retrospective study has been carried out, over a group of 234 patients, analysing comparatively their age, sex, environment background, occurrence circumstances and year season. RESULTS: The dominance of male patients can be remarked; according to their age, the dominance is between 40 and 55; according to the time period, the greater occurrence is between october and december. CONCLUSIONS: The study stresses out the similarity between the two groups (from BRASOV and TARGOVISTE): a greater occurrence of the male patients, the dominance of the perforant injuries of occular globe, the most affected age range is between 40 and 55. PMID- 12723181 TI - [Ocular biometry theory and practice]. AB - The paper presents elements of eye biometry theory and practice as seen by a young resident, in his first years of practice as ophthalmologist. The paper refers to the current stage of IOL measurement technique. Also, challenges for the younger generation are presented, such as new formulas for the patients who have undergone refractive surgery, measurement solutions for the patients affected by cataract, whose vitreous was replaced by silicone, and last but not least, the proper approach as regards biometrical measurements for children. PMID- 12723182 TI - [Correlation between iridology and general pathology]. AB - PURPOSE: The research proposal is to evaluate the association between certain irian signs and general pathology of studied patients. METHOD: There were studied 57 hospitalized patients; there was taken over all their iris images, which were analyzed through iridological protocols; in the same time the pathology of these patients was noted from their records in the hospital, concordant with the clinical diagnosis; all these information were included in a database for a computerised processing. RESULTS: The correlations resulted from, shows a high connection between the irian constitution establish through iridological criteria and the existent pathology. CONCLUSIONS: Iris examination can be very useful for diagnosis of a certain general pathology, in a holistic approach of the patient. PMID- 12723183 TI - [Management of intra-vitreous hemorrhage in diabetes]. PMID- 12723184 TI - [Trans scleral cyclophotocoagulation with diode laser, modern method of treatment in cyclodestructive surgery]. AB - PURPOSE: To analyze the results in lower IOP using cyclophotocoagulaton with diode laser in a refractory glaucoma series of cases (standard protocol). METHOD: 23 eyes from 23 subjects, watched for 6 months (preliminary results). Power 1600 mW, exposure 500 ms, wave 810 nm. The parameters who had been watched: IOP, VA, neovascularization, corneal oedema, epitelial breaks, pain and photophobia. RESULTS: IOP had changed from 32.08 mm Hg to 20.47 mm Hg. VA had been stationary in 65.2% cases; decreased in 30.43% and increased in 4.34% cases; 43.47% subjects needed just another laser session and 13.04% needed two another interventions. Average sessions 1.69; average power 18.08 W; average applications 22.6. CONCLUSIONS: The protocol used, repeated in need, seems to be sure and efficient in lower IOP and in ocular pain relief in refractory glaucoma cases with minimum VA. PMID- 12723185 TI - [The role of Brinzolamide 1% ophtalmic suspension in corneal edema]. AB - PURPOSE: The study is aimed at assessing the efficacy and safety of topical carbonic anhydrase inhibitor (IAC--i.e. ophthalmic suspension of brinzolamide 1%- Azopt) in the treatment of corneal edema. METHOD: For this purpose, we carried out an experimental, epidemiological-operational, randomised, placebo-controlled and double blind study, on three groups of laboratory animals (rabbits): group 1 (5 rabbits) included endothelial corneal injuries inflicted by ultrasound exposure by means of a phacoemulsification sound in both eyes group 2 (5 rabbits) included endothelial corneal injuries inflicted by direct trauma in both eyes group 3 (3 rabbits)--no endothelial corneal lesions (control group) In each group we instilled, all the animals, with ophthalmic suspension of brinzolamide 1% (Azopt), 1 drop t.i.d., in one eye for 14 days (starting the following day after infliction of endothelial injuries, when the corneal edema was evidenced by biomicroscopic examination) and placebo (saline solution 0.9% with the same osmolar and pH values as the Azopt) in the other eye. The assessment was performed throughout the following stages: To: preceding the infliction of corneal endothelial injuries, resorting to: biomicroscopic examination ultrasound pahimetry direct specular microscopy T1: the following day after infliction of corneal endothelial injuries, resorting to: biomicroscopic examination (the assessment of corneal edema) ultrasound pahimetry (measurement of corneal thickness and, hence edema) direct specular microscopy (cell density and endothelial ultrastructure evaluation: cellular polimegetism, pleiomorphism) T2: in the wake of 14 days therapy (after eye removal), adding to the previous examinations: indirect specular microscopy (with the same aim as the direct examination, yet examination carried out from the endothelial side) pathologic examination (using hematoxilin-eosin dye and Van-Gieson dye). PMID- 12723186 TI - [Posterior capsule rupture in lens surgery]. AB - PURPOSE: The paper studies the breaks of the posterior capsule in lens surgery, specifically the incidence, causes, types, management and the methods of IOL implantation. METHODS: For this purpose, a retrospective clinical trial was performed, within: 01.06.1999-31.05.2002, on 346 eyes group operated for senile cataract (without other ocular diseases) Patients were split in two groups, based on surgical technique: group 1 with 208 eyes operated using phacoemulsification and group 2 with 138 eyes operated using extracapsular extraction technique. RESULTS: The incidence of the breaks of the posterior capsule was 8.17% for group 1 (17 eyes out of 208) and 5.07% for group 2 (7 eyes out of 138). In all cases an IOL was used, most of them being placed in the posterior chamber (in bag, in sulcus or sclera-sutured) and infrequently IOL for anterior chamber or with irian support. CONCLUSIONS: Breaks of the posterior capsule is a complication of cataract surgery, but appropriate management (viscoelastic substances, anterior vitrectomy) leads to a good outcome, with no influence on the subsequent evolution. PMID- 12723187 TI - [The simultaneous contrast of the colors in Van Gogh paints]. AB - Vincent Van Gogh, Dutchman painter, is a forerunner but also a spokesman of the expressionism. His life was short and sad, marked by a psychic instability. In his paintings the color is subordinated of some subjective motivations. Van Gogh uses complementary colors to get a maximum intensity of the overlapped tones. The chromatic increase serves to obtain some strong emotions. In his landscapes the special distances are suggested through tonal distances. The increase of the pure tones through complementary contrast, the cancel of the imitation colors through sensation-colors, the explosive chromatic are characteristic of her painting. In his singular creation, Vincent Van Gogh created his suffering ego. PMID- 12723188 TI - [Contrast sensitivity]. AB - This paper presents a few theoretical considerations about the contrast sensitivity, modalities of testing and changes of contrast sensitivity in different ophthalmologic diseases. PMID- 12723189 TI - [Pseudotumor cerebri or idiopathic cranial hypertension. Neuro-ophthalmologic considerations]. AB - Pseudotumor cerebri is defined and the general pathology added where the idiopathic intracranial hypertension syndrom can be found. The paper reviews the consequences that the increase in cerebral spinal liquid (CSL) pressure has on the visual function. The symptomatology is polymorphous, hard to frame and much more difficult to cure, the diagnosis being one of exclusion. PMID- 12723190 TI - [Optic nerve bilateral atrophy in a suprachiasmatic astrocytoma]. AB - The paper presents a clinical case of a 5 years old child with bilateral optic atrophy and the specific diagnosis and treatment problems of this case. The image investigations have a major part in establishing the positive diagnosis: computerized tomography, magnetic resonance imaging and the elective treatment belongs to the field of neurosurgery. PMID- 12723191 TI - [Post-inflammatory glaucoma--methods of treatment]. AB - We show the mechanisms of postinflammatory secondary glaucoma and the possibilities of treatment of this, all of this concerning to an individual case. We evaluate the benefits and risks for every kind of therapeutical approach and we show our choice. PMID- 12723192 TI - [Therapeutical difficulties in a patient with TB panuveitis and open angle glaucoma]. AB - The article present the case of a young female patient showing TB panuveitis associated with primary glaucoma, as well as all the implications of the symptomatic and ethiological treatment of TB panuveitis on the increased intraocular pressions (IOP). There are also underlined the difficulties of maintaining the IOP between normal ranges under combined therapy for glaucoma. PMID- 12723193 TI - [Diabetic retinopathy in newly diagnosed patients with type II diabetes mellitus]. AB - PURPOSE: To report the prevalence of the diabetic retinopathy in newly diagnosed patients with type II diabetes mellitus and to evaluate the association between the retinopathy and different clinical variables. METHOD: A prospective study including all the newly diagnosed patients with type II diabetes mellitus between January 2001-March 2002 in the evidence of the Diabetes Center from Cluj has been conducted. The fundus appearance, age, sex, incidence of arterial hypertension and the diabetes therapy (diet alone, oral drugs, insulin) have been evaluated. RESULTS: During January 2001-March 2002, 487 patients have been diagnosed with type II diabetes mellitus in Cluj county. Diabetic retinopathy has been identified in 70 of these patients (14.37%). Most of the newly diagnosed patients with type II diabetes mellitus belonged to the age interval: 40-65 years, with an average of 58-59 years. CONCLUSIONS: Among the 70 patients with diabetic retinopathy at the moment of type II diabetes diagnosis, the first place is occupied by the back-ground diabetic retinopathy. (11.70%), followed by the clinically significant macular edema (1.43%), proliferative diabetic retinopathy (1.02%) and preproliferative diabetic retinopathy (0.20%). Systolic and diastolic arterial hypertension has been associated with higher degrees of severity of the diabetic retinopathy. PMID- 12723194 TI - [Primary open angle glaucoma evolution after surgery]. AB - The authors present a four year (01.02.1998-01.02.2002) clinical retrospective trial, including 209 eyes operated for Primary Open Angle Glaucoma (POAG). 52 eyes (24.8%) out of 209 eyes the progression of the disease was discovered observing the increase of the intraocular pressure (IOP), failure in achieving "IOP target" due to the insufficient filtration, the obstruction of the filtration and the deterioration of the papiloperimetrics parameters. The therapeutic approach consisted of additional medication, filtration revision, subsequent surgical treatment physical end/or classic. The authors suggest a sequence for the operated cases of POAG, but that continued to progress, postoperative IOP "seemingly normal" requiring the follow-up for t e disease, in spite of using entire methods of the treatment. PMID- 12723195 TI - [Particular aspects of treatment in diabetic eye cataract]. AB - The knowing of the ocular consequences of diabetes is essential for the evaluation of the risk factors for the cataract surgery. The clinical study was made on a total number of 19 diabetic patients (3 with insulin dependent diabetes and the rest with noninsulin dependent diabetes), which were operated for cataract from september 2001 till march 2002. Preoperatory preparation presumed a good metabolic control of diabetes and a very carefully examination of the retina. The surgical technique was the extracapsular cataract extraction, followed by the implantation of a posterior chamber IOL, respectively phacoemulsification with the implantation of a foldable lens. The presence of a proliferative diabetic retinopathy, of a rubeosis iridis, of the neovascularization of the angle, or the preexistence of the vitreoretinal surgery (vitrectomy) were the main factors which dictated the anatomical and functional postoperatory prognosis. In conclusion, evenly the diabetic patient is a highly risk patient, the adequate preoperator preparation, the right treatment and a carefully follow-up could assure the success of the surgery in those cases. PMID- 12723196 TI - [Optic nerve lesions in diabetes mellitus]. AB - The present paper is a retrospective study revealing some optic nerve lesions on a group of 1.700 diabetic patients which belonged to the common casuistry of the Ophthalmologic Clinic and Diabetic and Nutrition Diseases Clinic in Craiova. A reduced frequency of optic neuropathy in diabetic persons was established such as: 27 patients (1.58%). Affected optic nerve may present various clinical aspects among which ischemic optic neuropathy is predominant (59.20%) followed by secondary optic atrophy, ischemic optic post neuropathy (33.40%) and juxtabulbar optic neuritis (7.40%). PMID- 12723197 TI - [The incidence of diabetic retinopathy on a set of 10,000 patients with diabetes mellitus]. PMID- 12723198 TI - [Is atropine efficient in para-bulbar injections?]. AB - Supposing the fact that atropine has antispastic effect on blood vessels, we studied the variation of blood flow in ophthalmic artery, central retinal artery and ciliary arteries, by Doppler ultrasound. We used 1 ml Atropine 1@1000 in parabulbar administration to 17 eyes with neuro-retinal diseases and the arterial flow was registered for 3 times: before the injection, at 15 and respective 30 minutes after administration. The results were processed using t-Student method. The atropine effect seemed to be more intensive 30 minutes after administration on central retinal artery. PMID- 12723199 TI - [Immunopathologic mechanism in some ocular diseases]. AB - OBJECTIVES: The goal of the present study is to elucidate the pathogenic mechanisms that determine the ocular involvement in some systemic diseases. MATERIAL AND METHODS: We have studied 45 patients with ocular diseases and concomitant systemic diseases (systemic lupus erythematosus-SLE, ankylosing spondylitis, Reiter syndrome, juvenile rheumatoid arthritis, Sjogren syndrome). The patients studied have been compared with a control group represented by 20 healthy individuals. In each case we have determined the serum levels of the following humoral markers: the C3 complement fraction, circulating immune complexes, autoantibodies (antinuclear autoantibodies, anti-double stranded DNA autoantibodies, anti-smooth muscle autoantibodies, anti-neutrophilic cytoplasm autoantibodies, anti-cardiolipin autoantibodies, anti-SSA, anti-SSB, rheumatoid factor). RESULTS: The autoantibodies detected in certain cases of uveitis suggest the existence of an autoimmune etiology; the antibody titers have been high in some severe cases of rheumatic uveitis, especially in patients with ankylosing spondylitis. We have found that there is a correlation between the presence and the titer of certain autoantibodies, and the onset and evolution of the disease. Specifically, the antinuclear autoantibodies may be considered a predictive factor for the development of chorio-retinopathy of SLE. Uveitis mediated by immune complexes has been seen in rheumatoid arthritis, ankylosing spondylitis, and SLE, these being the same systemic diseases where there is an important pathogenic role of immune complexes. CONCLUSIONS: The abnormal immunologic function, that was observed in the ocular inflammatory diseases studied, may be synthesized by the association between two immunopathologic phenomena; both of them are finalized by the non-self transformation of the antigens linked to various eye structures. These are represented by autoimmunity (of humoral, cellular, or mixed type), and the immediate type III hypersensitivity reaction. This study provides important data regarding the involvement of autoimmune diseases in the etiology of ocular diseases. PMID- 12723200 TI - [Humoral and cellular immunity investigation in idiopathic uveitis]. AB - OBJECTIVES: There is an amount of controversy regarding the patho-physiology of uveitis. Due to the fact that the eye (and especially the highly vascular tissues such as the uvea and the conjunctiva) is considered a special "target" for the immunopathologic reactions, the goal of our study is to contribute to the understanding of the causal role of the immune reactions in the development of idiopathic uveitis. MATERIAL AND METHODS: We have studied 48 patients with idiopathic uveitis and 20 control subjects (with no ocular symptomatology) for the following humoral and cellular immune markers: IgM and IgG, the C3 complement fraction, circulating immune complexes, autoantibodies (antinuclear autoantibodies, anti-double stranded DNA autoantibodies, anti-smooth muscle autoantibodies, anti-neutrophilic cytoplasm autoantibodies, anti-cardiolipin autoantibodies, anti-parietal cell autoantibodies), and lymphocyte subtypes. RESULTS: Changes in the serum levels of the immune humoral and cellular markers have been noted in 39 (81.25%) of the patients studied. C3 was decreased in 12 (30%) of all cases; the immune complex level was increased over 50 U/mL in 20 patients (51.28%). The presence of the above-mentioned autoantibodies was seen in 19 patients (48.71%), with different incidence. By flow cytometry the percentage of CD4+ (T helper lymphocytes) has been found to be decreased, and the percentage of CD8+ was found to be increased in 8 cases (20.51%). The opposite, the increase in the percentage of CD4+ lymphocytes, and the decrease in the percentage of CD8+ cells was seen in 25 (64.10%) of the studied patients. Taking into consideration the clinical symptoms and the level of the immune humoral and cellular parameters, the cases included in the study have been included in two groups: anterior uveitis (acute and chronic), and posterior uveitis. CONCLUSIONS: By comparing the results, we conclude that the autoimmunity and the immediate hypersensitivity mediated by immune complexes may be involved in the appearance and the development of the intraocular inflammation in certain cases of idiopathic uveitis. PMID- 12723202 TI - [Internal tamponade in proliferative diabetic retinopathy surgery]. AB - Vitreous hemorrhage has been the most common indication of posterior vitrectomy in diabetic retinopathy. Now, the leader is tractional retinal detachement and many others: rhegmatogenous retinal detachement, fibrovascular proliferations membranes, huge premacular hemorrhage. During vitrectomy procedure it may be necessary internal tamponade using air, expanding gases, silicon oil and/or perfluorocarbon liquids. PMID- 12723201 TI - [Acute posterior multifocal placoid pigmentary epitheliopathy--case report ]. AB - We present a case of young female diagnosed with acute posterior multifocal placoid pigment epitheliopathy, presented with decreased visual acuity. Main differential diagnosis were discussed as well as some general data regarding the pathology of this relatively rare syndrome. PMID- 12723203 TI - [Secondary cataract treatment with laser YAG-ND]. AB - The purpose of the study is to present the favorable results of YAG: Nd laser capsulotomy for the treatment of secondary opacification of the posterior capsule, in our department. A 3 years retrospective study was performed. For 1440 cataract operations, most of them extracapsular extractions with PMMA IOL placed in the posterior chamber (ciliary sulcus or capsular bag), 285 laser capsulotomies were done. The results are presented statistically, including sex and age distribution, type of cataract surgery and implant, treatment protocol (number of sessions, number of impulses per session, energy per impulse) and complications. The frequency of secondary capsular opacification in our study is 19.78%. It is most important in the age group 60-70 years, where the surgery for cataract is performed more often. Capsulotomy was done after extracapsular extraction with IOL placed in the posterior chamber, but especially after extracapsular extraction without implant placement or with complications that have permitted only the use of an anterior chamber implant. Capsulotomy is performed in only one session in 88% of the cases. The energy per impulse is usually 3-5 mJ and the mean number of impulses per session is 10-30. There are few complications: transient elevation of intraocular pressure in 14.73%, discrete laser marks on the implant in 8.77% and moderate iritis in 1% of the cases. YAG: Nd laser capsulotomy appears to be a safe and efficient method of treating secondary cataract; it improved visual acuity in all cases, if different ocular pathology had not influenced it. PMID- 12723205 TI - [Temporary sutured trabeculotomy in primary open angle glaucoma]. AB - The paper presents the results in surgical treatment of open angle glaucoma with a removable suture trabeculectomy technique. 25 eyes of 19 patients with different stages of open angle glaucoma were operated; 7 eyes had pseudoexfoliation glaucoma. The results obtained on this patients were compared to those obtained on the patients operated with classical trabeculectomy. In the final we discuss also the results presented by the Italian authors. Preoperative medium IOP was 33.04 mmHg +/- 13. The pressional reduction was over 50% on the patients with preoperative values of IOP under 30 mmHg, and over 70% on the patients with preoperative values of IOP over 30 mmHg. The follow-up period was 6 months. In the present all patients have IOP in normal ranges without additional medication. PMID- 12723204 TI - [Complications at the beginning of refractive surgery]. AB - We revised the charts of first interventions of refractive surgery with LASIK and PRK in order to evaluate the results and to analyze the incidents and complications we encountered as beginners in photorefractive surgery. In total 50 eyes of 29 patients (7 men and 22 women) were operated. 42 eyes were myopic (21 with astigmatism) and 8 eyes were hyperopic (6 with astigmatism). LASIK was performed in 45 cases and PRK in 5 cases all myopias. The intervention was performed with AESCULAP MEDITEC MEL 70G-Scan laser. The follow-up period was between 3 and 12 months. Per total the results were as follows: Average pre operative non-corrected visual acuity was 0.23 Average pre-operative corrected visual acuity was 0.8 Average post-operative non-corrected visual acuity was 0.74 Average post-operative corrected visual acuity was 0.77 We noted some minor intra operative incidents: insufficient anesthesia (6%) and some important intra operative incidents: free cap (6%), flap desepithelization (4%). Minor post operative complications were: mild corneal edema, corneal desepitelisation (14%), and some severe post-operative complications: corneal ulcer (4%), comeal mycosis (2%), pupilary paresis (2%). Under-correction was present per total in 52% of cases, but mainly in myopias over 10 dpt (30%), as well as induced astigmatism (10%). Major complicated cases were only 5 (10%) and were consecutive to some mechanical problems (vacuum failure, system decentration). An interesting observation is related to IOP evolution. In average a decrease of IOP was noted from pre-operative medium of 14.43 mmHg to a post-operative medium value of 10.73 mmHg. PMID- 12723206 TI - [Trichromatic method Gomori in anatomopathologic study of ocular structure]. AB - Histologic sections performed from the anterior and posterior pole of the eye were stained by the trichrome method described by Gomori. There are described details of the cornea and ciliary body in normal and pathologic conditions. There are discussed advantages of this method over routine stained slider, especially in the case of achromic melanoma. PMID- 12723207 TI - Photodynamic occlusion of ocular neo-vascularization with B2 vitamin. AB - Photodynamic therapy (PDT) of cancer has been known for over twenty years and is based on the dye-sensitized photooxidation of different biological targets in the tumoral tissue yielding to a photochemically induced cell's death via apoptotic pathways. Several parameters affect clinical trials in PDT and influence the therapeutic outcome. A potentially major application of PDT in a non-cancer field is its use in treatment of age-related macular degeneration. This condition, caused by proliferation of neovasculature in the retina, is the major cause of blindness in the over 50s. Using a photosensitiser which enters neovasculature very rapidly following administration and is subsequently quickly moved from the circulation. Ocular neovascular disease represents an important cause of blindness today. In this paper, was used an animal model (rabbit) to evaluate the vaso-occlusive potential of photodynamic therapy with B2 vitamin. Neovascularization induced in the rabbit cornea consists of an easily accessible monolayer-like neovascular net within a transparent matrix. This fact allows studying the efficiency of occlusion in an isolated neovascular structure. PMID- 12723208 TI - [Arteries of visual cortex]. AB - The purpose of the study was to point out the arterial branches that are supplying the visual cortex. This study was carried out on 100 brains, fixed in formaldehid 10% and dissected under the surgical microscope; 20 of them were at first injected with colored nitrolac. It was observed the origin, relationships and distribution of the arterial branches, which supply these areas. The data obtained reveal that: medial face of the visual cortex receives branches from the posterior cerebral artery (throughout its collaterals: calcarine artery 100%, parietooccipital artery 96% and posterior temporal artery 92%; its lateral surface receives cortical branches from the middle cerebral artery (100%) and its inferior surface is irigated from collaterals of the posterior temporal and temporo-occipital arteries (100%). Some of the branches penetrate directly the visual cortex perpendiculary and others form at first, a superficial, pial network. The knowledge of arterial variants which supply the visual areas is very important for ophthalmologists, radiologists, neuroophthalmologists, neurosurgeons, for recognising their vascular damage in some complex neuroophthalmologic syndroms. PMID- 12723209 TI - Perioperative medical considerations for the geriatric dental patient. Risk assessment and management. PMID- 12723210 TI - Managing pain for the elderly. PMID- 12723211 TI - Considerations for dental treatment in long-term care facilities. PMID- 12723212 TI - Experience-based dentistry in the nursing home setting. PMID- 12723213 TI - Caries prevalence, nutrition, and xerogenic medication use among a geriatric population. PMID- 12723214 TI - A clinico-pathologic correlation. Condensing osteitis (focal sclerosing osteomyelitis). PMID- 12723215 TI - [Reliability of the glutaraldehyde test to measure gamma-globulin levels in foals and the use of this test to check colostrum intake of foals]. AB - The glutaraldehyde coagulation test is a semi-quantitative test used to determine the gammaglobulin concentration in serum. The purpose of this study was to examine the reliability of the different modifications of this test by determining the sensitivity, specificity, positive and negative predictive value, and the prevalence of hypogammaglobulinemia in foals. The results of the glutaraldehyde coagulation test were compared with the serum gammaglobulin concentration as a reference value, determined by measuring total serum protein and the serum protein spectrum. It was concluded that the glutaraldehyde coagulation test is a suitable test to use in the field for determining the serum gammaglobulin concentration in foals. The test has good sensitivity, specificity, positive and negative predictive value when using a 5% glutaraldehyde solution and when results are evaluated at 30 minutes for a serum concentration of 8 g/l and at 50 minutes for a serum concentration of 5.5 g/l, a concentration below which therapy is recommended. PMID- 12723216 TI - [Active care!]. PMID- 12723217 TI - [Interview with Susan E. Shaw at Chateau Veterinair. Dangers of traveling with pets through Europe. Interview by F. J. W. C. van Herten]. PMID- 12723218 TI - [The Regulation of Aggressive Animals and the Staffordshire terrier]. PMID- 12723219 TI - [Priapism in the stallion and in man]. PMID- 12723220 TI - [Veterinary homeopathy at war with the Code for Veterinarians]. PMID- 12723221 TI - [College called to order..!]. PMID- 12723222 TI - [Substitute veterinarian on the pay role]. PMID- 12723223 TI - Your practice in the palm of your hand. PMID- 12723224 TI - Dentists talking negatively about dentists. PMID- 12723225 TI - Why not prosthodontists? PMID- 12723226 TI - Oral pathology quiz #38. Case 1. Pyogenic granuloma. PMID- 12723227 TI - Oral pathology quiz #38. Case 2. Mucoepidermoid carcinoma. PMID- 12723228 TI - Oral pathology quiz #38. Case 3. Ameloblastoma. PMID- 12723229 TI - Oral pathology quiz #38. Case 4. Odontogenic keratocysts. PMID- 12723230 TI - Low level mercury exposure in dentists--the tip of the iceberg. PMID- 12723231 TI - Staying hip to HIPAA! PMID- 12723232 TI - The best Christmas gift ever: the day I got the call. PMID- 12723233 TI - The healing paradox. PMID- 12723235 TI - [Several aspects of young students' health]. AB - The authors calculated probable risks of main pathologic syndromes, considering whole motivational sphere and personal features of future employees engaged into aircraft building and chemical industries. PMID- 12723234 TI - [Use of simulation experiments to forecast the development of vibration disease]. AB - The article covers results of simulation regression experiments that provided a intensity-time-effect dependence for longevity, vibration dose and parameters of endocrine functions. PMID- 12723236 TI - [Altered immunologic reactivity in pregnant women living in ecologically unfavorable conditions]. AB - Pregnant women living in major industrial center near Baikal were proved to have disordered immune response associated not only with disbalance of helpers and suppressors, but also with altered activation of mononuclear phagocytes and T helpers. The authors revealed relationships between genetic structures of HLA complex and activity of various links in immune system. PMID- 12723237 TI - [Toxicologic and hygienic aspects in evaluating the risk due to isolated and combined action of biotechnologic products]. AB - The article covers topics of evaluating the risk caused by biotechnologic products action both isolated and combined with chemical factor. The biotechnologic products under study were complete proteous preparations and microorganisms producers of nutritive supplements. The authors revealed peculiarities and patterns of body response to biotechnologic synthetic products. PMID- 12723238 TI - [Evaluation of health changes in children under environmental hazards]. AB - The authors considered changes of biochemical parameters of thyroid, lipid and proteous metabolism in children under chronic multicomponent exposure to ambient air pollution along with moderate iodine deficit as a result of interaction between intoxication processes and human compensatory reactions. PMID- 12723239 TI - [Characteristics of work conditions and evaluation of health status of firemen]. AB - Hygienic evaluation of fireman's work conditions revealed that smoke generation zone near ignition source contains a complex of toxic chemicals in concentrations hundreds of times exceeding the MACs. Staffers are exposed to noise and vibration, levels of which exceed the normal values. Those who more frequently participated in fire extinguishing and therefore faced more toxic factors demonstrate higher levels of disablement. PMID- 12723240 TI - [Progression of toxic encephalopathy in firemen]. AB - The authors analyzed possible causes of encephalopathy progression in late occupational intoxication among firemen. Those causes are disordered autoregulation of cerebral vessels, oxidant and psychoemotional stress, autoimmune disorders. PMID- 12723241 TI - [Disorders of main stress effector parameters under effects of local vibration]. AB - The authors studied some parameters of hormonal, immune and cytokine state in workers exposed to vibration. Considering local vibration as a chronic stressful factor, the authors analyzed changes in the studied interplaying neuroendocrine and immune parameters in human response to physical stress. PMID- 12723242 TI - [Methodical approaches to determination of microelements in biologic materials]. AB - The authors present data on sensitivity, accuracy and reliability of analytic methods suitable for mass determination of major microelements in human biologic materials. The findings are that, considering probable microelements content, combination of atomic absorptive analyzers appears optimal. PMID- 12723243 TI - [Evaluation of the state of 2d phase of xenobiotics biotransformation in firemen who participated in fire extinguishing at AO "Irkutskkabel"]. PMID- 12723244 TI - [Use of mathematical models to evaluate influence of environmental air pollution on children's health]. PMID- 12723245 TI - [Course of encephalopathy in the late period of occupational mercury intoxication]. PMID- 12723246 TI - Fever after a stay in the tropics. Part 1: Diagnostic approach. PMID- 12723247 TI - Fever after a stay in the tropics. Part 2: Common imported tropical diseases. PMID- 12723248 TI - Outpatient and home parenteral antibiotic therapy (OHPAT) in low-risk febrile neutropenia: consensus statement of a Belgian panel. AB - Febrile neutropenia requires adequate antibiotic treatment. A subgroup of patients are only at low risk for complications and could be treated at home/as outpatients (OHPAT) after a short initial admission for work up. This position paper by a Belgian panel of experts presents criteria defining low-risk in febrile neutropenia, gives an overview of the existing experience and examines the present obstacles to a more widespread use of OHPAT in this country. PMID- 12723249 TI - Autoimmunity and pregnancy: theory and practice. PMID- 12723250 TI - Plasma D-dimer concentrations in different clinical conditions. AB - D-dimers (DD), specific degradation products of crosslinked fibrin, are markers for activation of plasma coagulation and/or fibrinolysis. DD results below the cut-off level are proven to be useful to rule out the probable diagnosis of deep venous thrombosis (DVT) and/or pulmonary embolism (PE). Our objective was to demonstrate that positive DD occur in many diseases and certain physiological conditions as high age and pregnancy and to look for gradations in positivity between different pathological conditions. We wanted to investigate the request attitude of our clinicians concerning DD. Positive DD results still confuse some physicians. Retrospectively, we examined medical records of 574 consecutive patients, in whom plasma DD were measured in daily routine. Both outpatients (n = 423) and inpatients (n = 151) were included. We noted their clinically predominant disease. Measurement of DD concentration is too often requested by clinicians, in any medical condition, and is not always clinically relevant. The relation of a positive result and the clinical problem is sometimes not understood. Overall, we found 64% DD positivity with a median concentration of 775 micrograms/L. We found elevated DD concentrations in various clinical conditions. PMID- 12723251 TI - Thromboprophylaxis in general medical patients. Recommendations of the Thrombosis Guidelines Group of the Belgian Society on Thrombosis and Haemostasis and the Belgian Working Group on Angiology. PMID- 12723252 TI - Surgical treatment for pulmonary metastases. AB - As the lungs are a filter for the entire circulation, they are a common site for metastatic disease. Although never proven by a prospective randomized trial, surgical resection is a widely accepted treatment for pulmonary metastases on the condition that a complete resection can be obtained. However, many patients will develop recurrent disease in the thorax despite the use of systemic chemotherapy, dosage of which is limited due to systemic toxicity. As isolated limb and liver perfusion, isolated lung perfusion is an attractive and promising surgical technique for the delivery of high-dose chemotherapy with minimal systemic toxicity. The use of biological response modifiers, like tumour necrosis factor, is also feasible. Isolated lung perfusion has proven to be highly effective in rat models of pulmonary metastases with a clear survival advantage. Lung levels are significantly higher after isolated lung perfusion compared to intravenous therapy without systemic exposure. Preliminary human studies have shown that isolated lung perfusion is technically feasible with low morbidity and without compromising the patient's pulmonary function. Further clinical research is necessary to determine its effect on pulmonary metastases in man, especially in case of unresectable disease or as adjuvant therapy after surgical resection. PMID- 12723253 TI - Paralysis due to severe hypokalemia. AB - We report two cases of a patient with paresis/paralysis caused by a severe iatrogenic hypokalemia. In both cases, the extreme muscle weakness disappeared with correction of the electrolyte disturbance. The mechanisms of hypokalemia due to nephrotoxic drugs and treatment of this electrolyte disturbance are discussed. PMID- 12723254 TI - Primary meningococcal arthritis of the hip in an immunocompetent adolescent. AB - The clinical spectrum of meningococcal infection ranges from asymptomatic carriage to fulminant sepsis, with meningitis and septicemia being well recognized clinical presentations. Meningococcal arthritis as a complication of Neisseria meningitidis infection occurs in about 2-10% of cases, whereas primary meningococcal arthritis (PMA) is a relatively rare phenomenon, even in children. We report here a case of meningococcal infection in an immunocompetent adolescent suffering from acute pain of the right hip as the only symptom upon presentation at the hospital. In such a situation, meningococci are not usually considered as a possible causative agent. PMID- 12723256 TI - Genetic fibrillinopathies: new insights in molecular diagnosis and clinical management. AB - The Marfan syndrome (MFS) is an autosomal dominant connective tissue disorder with a prevalence of 2-3 per 10,000 individuals and symptoms ranging from skeletal overgrowth, cutaneous striae to ectopia lentis and aortic dilatation leading to dissection. Mutation in the gene for fibrillin-1 (FBN1) cause MFS and other related disorders of connective tissue, grouped as fibrillinopathies. Fibrillin-1 is the main constituent of extracellular microfibrils. Microfibrils can exist as individual structures or associate with elastin to form elastic fibers. This article provides an overview of the current diagnostic criteria and medical management, estimates the role of fibrillin-1 mutation analysis, sheds new light on genotype-phenotype correlations and summarizes new insights on the pathogenesis of this disorder based on mouse models. PMID- 12723255 TI - Biliary complications of large Echinococcus granulosus cysts: report of 2 cases and review of the literature. AB - Hydatid cysts are often incidentally found and remain clinically silent. However complications can occur. We present 2 patients who developed biliary complications due to a large hydatid cyst. In the first patient compression on the intrahepatic bile ducts and cystic duct by the cyst, caused cholangitis and cholecystitis. Moreover the cyst had ruptured into the right intrahepatic bile ducts. A sphincterotomy was performed with extraction of hydatid sand. A pericystectomy was necessary because of infectious deterioration of the patient. Albendazole was continued for 8 weeks after surgery. The second case presented with jaundice and weight-loss since 1 month. A large hydatid cyst caused compression on the bile duct bifurcation with proximal bile duct dilatation. A cystectomy was performed 2 weeks after albendazole therapy initiation, which was continued for 8 weeks after surgery. Follow-up of both surgical interventions was unremarkable. Although Echinococcus granulosus in not prevalent in Belgium, we must be aware of this pathology in patients coming from high endemic regions. PMID- 12723257 TI - [Severe diarrhea due to rotavirus infection in a Belgian hospital 1981-2002]. AB - Rotavirus infections are a major cause of severe diarrhea in children younger than 2 years. In Belgium they cause many hospitalizations because of dehydration. A study of the laboratory diagnosis of rotavirus infections in 28.251 stool samples at a university teaching hospital in Belgium during a twenty-year period (1981-2002) showed a marked seasonality. The virus was most often diagnosed during the winter months: 54% of the rotavirus isolates were found in the first three months of the year, with 21% of the positive samples occurring in February. Recently, rotaviruses can be genotyped based on differences in the viral outer capsid protein VP7. Vaccines are currently being developed against the four most prevalent genotypes G1, G2, G3 and G4. During the last three epidemic seasons (1999-2002) in Belgium, G1 was the most prevalent genotype and accounted for 62% of the rotavirus isolates recovered. G2, G3 and G4 were also isolated, and other emerging types need to be carefully monitored too, since G9 (45%) was co-dominant with G1 (42%) in the 2000-2001 rotavirus season in Belgium. The future development of an efficient rotavirus vaccine will need to take the diversity of the rotavirus genotypes into account. PMID- 12723258 TI - Antibiotic susceptibility and serotype distribution of 240 Streptococcus pneumoniae causing meningitis in Belgium 1997-2000. AB - Cerebrospinal fluid isolates of Streptococcus pneumoniae, collected during the years 1997-2000 at more than 100 Belgian laboratories were studied. The 10 most common serotypes-serogroups representing 76% of the isolates were 14, 6, 9, 19, 23, 18, 4, 10, 8 and 12 (in order of frequency). Thirty-six percent of strains were isolated in children < 5 years old. In this age group the number of serogroups was more limited and 81.4% are included in the 7-valent conjugate vaccine. Decreased susceptibility to penicillin was observed in 13.9% of 237 strains (MIC > 0.06 mg/L), with only 2.1% resistant strains (MIC > 1 mg/L). Twelve strains showed reduced susceptibility to cefotaxime (MIC > 0.5 mg/L). Only three of the 237 strains were intermediately susceptible to meropenem. All strains were susceptible to vancomycin and moxifloxacin. In Belgium, high doses of third generation cephalosporins remain effective for the treatment of pneumococcal meningitis. The new fluoroquinolones seem the most promising agents for the treatment of pneumococcal meningitis in the future. PMID- 12723259 TI - Is there a rationale for prescription of benzodiazepines in the elderly? Review of the literature. AB - Benzodiazepines (BZDs) constitute the most widely used symptomatic treatment of insomnia and anxiety. Many of these drugs are associated with adverse effects, such as daytime sedation and dependence with continued use. There is a concern about the rationale for and extent of benzodiazepine (BZD) use in the elderly. The sedation due to BZD use is a main risk factor for falls and other accidents. Impaired cognitive function with continuous use appears to be a major side effect. There is a general awareness that BZD use is inappropriate in many patients, and therefore discontinuation should be recommended whenever possible. Moreover, long-term use of these drugs should be actively discouraged. Although no unanimous recommendations concerning the optimal duration of the withdrawal process exist, BZDs may easily be withdrawn during a short period in most patients who are habituated to a low dose, if an initial phase with dose reduction and psychological support are provided. Alternative approaches involve sleep hygiene guidelines, behavioural treatment and psychotherapy tailored to the needs of the individual patient. PMID- 12723260 TI - Minitransplants: allogeneic stem cell transplantation with reduced toxicity. AB - Allogeneic hematopoietic stem cell transplantation (HSCT) is used for the treatment of selected hematological malignancies. Its curative potential is based on two very different mechanisms, involving the conditioning regimen and the graft-versus-host reactions, respectively. The high-dose chemo-radiotherapy conditioning regimen is aimed at destroying tumor cells, ablating the host immune system (to prevent rejection) and eliminating the host bone marrow (to "make space" for donor stem cells). However, the definitive eradication of tumor cells is also largely mediated by an immune-mediated destruction of malignant cells by donor lymphocytes termed graft-versus-leukemia (GVL) or graft-versus-tumor (GVT) effect. However, because of its toxicity, conventional allogeneic HSCT is restricted to younger (< 55 years) and fitter patients. These observations led several groups to set up new (less toxic) transplant protocols based on a two step approach: first the use of immunosuppressive (but nonmyeloablative) conditioning regimens providing sufficient immunosuppression to achieve engraftment of allogeneic hematopoietic stem cells and, in a second step, destruction of malignant cells by the GVL effect. These transplants are called nonmyeloablative HSCT or reduced-conditioning HSCT or minitransplants. Preliminary results show that minitransplants are feasible with a relatively low transplant-related mortality (TRM) even in patients up to 70 years. In addition, strong anti-tumor responses are observed in several hematological malignancies as well as in some patients with renal cell carcinoma. As the benefits of minitransplants over alternative forms of treatment remain to be demonstrated, this strategy should be restricted to patients included in clinical trials. PMID- 12723262 TI - [Minocycline-induced pulmonary infiltration and eosinophilia]. AB - A 90-year old woman with pulmonary infiltrates and eosinophilia caused by minocycline is described. Until now, in the English, French and Dutch literature only some 35 cases are known. The prognosis is good, especially after treatment with corticosteroids. This doesn't legitimate, however, the use of minocycline without a clear indication. This case again stresses the need for a careful history taking, especially regarding the intake of medications (like minocycline), when one is confronted with a diagnostic problem of pulmonary infiltrates and eosinophilia. PMID- 12723261 TI - Toxic intrathoracic goiter and mediastinal lymphadenopathy: an unusual presentation of systemic primary AL amyloidosis. AB - Mediastinal lymphadenopathy and goiter have been associated with primary amyloidosis, although not in the same patient. One previous case report described the association of an amyloid goiter and hyperthyroidism (due to Graves' disease) with primary amyloidosis. Till now no case reports of patients presenting simultaneously with mediastinal lymphadenopathy, intrathoracic amyloid goiter and hyperthyroidism as the first manifestation of systemic primary (idiopathic) amyloidosis have been described. The present case report describes the clinical, biological radiological and histological features in such a male patient. PMID- 12723263 TI - [Familial hypertrophic cardiomyopathy associated with Wolff-Parkinson-White syndrome]. AB - Familial hypertrophic cardiomyopathy (HCM) with Wolff-Parkinson-White (WPW) syndrome is extremely rare and associated with a high risk of ventricular tachyarrhythmia and sudden death. We report a familial form of hypertrophic cardiomyopathy associated with Wolff-Parkinson-White syndrome in two siblings 7 and 12-year-old. These patients showed progression to left ventricular dilatation. Early recognition and treatment of such forms can improve such evolution and the risk of sudden death. PMID- 12723265 TI - Notice of possible bylaws change. PMID- 12723264 TI - Extensive acronecrosis as a manifestation of mixed cryoglobulinaemia: a case report. AB - Cryoglobulinaemia is a systemic disorder characterized by circulating antibodies that precipitate in the cold and resolve on rewarming. Three different types have been described, distinct in the class of immunoglobulins and their clonality. The clinical expression varies from purpura and arthralgia to progressive renal failure and even acronecrosis (1-3). Associated conditions are lymphoproliferative disorders, auto-immune diseases and chronic infections, but several cases occur in the absence of identifyable other disease states. The present communication reports on a case of mixed cryoglobulinaemia. Of particular interest are the rapidly progressive clinical evolution to acronecrosis of the four limbs, necessitating amputation, the presence of spurious leucocytosis and the absence of other systemic symptoms. PMID- 12723266 TI - Testing your diagnostic skills. Case no 1. Malignant lymphoma. PMID- 12723268 TI - Laura Jamison to highlight practice management, communication skills. PMID- 12723267 TI - Testing your diagnostic skills. Case no. 2. Central giant cell lesion. PMID- 12723269 TI - Drs. Gremillion, Dolwick to present FNDC's first dissection courses. PMID- 12723270 TI - Whose info is it, anyway? As HIPAA provisions take effect, patients may be surprised by the level confidentiality, intent on exercising new rights. AB - As the long-awaited HIPAA privacy regulations finally take effect this week, the question remains as to just how warmly consumers will respond to the raft of new federal rules governing control of medical information. The adjustment period may be lengthy for these complex regulations, but the new rules certainly will educate patients about their rights as consumers, says consultant Jill Callahan Dennis, left. PMID- 12723271 TI - Barbakow exits as chairman, Leader will keep CEO title as Tenet Healthcare Corp. struggles to bring in new investors, send a message. PMID- 12723272 TI - Barely afloat. HealthSouth scrambles to avoid bankruptcy filing. PMID- 12723273 TI - Polling questions. Gallup accuses Scully of intimidation, collusion. PMID- 12723274 TI - Don't mess with VHA. Despite turmoil, Novation won't lie down for GPOs. PMID- 12723275 TI - Dodging a bullet. Rulings limit punitive damages, uphold arbitration. PMID- 12723276 TI - One insurer's plan to cover everyone. Universal coverage requires universal sacrifice by all segments of society. PMID- 12723277 TI - Universal appeal. Although there's strong consensus on the need to provide healthcare coverage for all Americans, that's where the agreement ends. PMID- 12723278 TI - Front and center. Dems making universal coverage a rallying cry. PMID- 12723279 TI - Targeting the middlemen. Pharmacy benefits managers under fire for prices. PMID- 12723280 TI - A '100 top hospitals' addendum. Statement from Solucient. PMID- 12723281 TI - WHO recommended measures for persons undertaking international travel from areas affected by severe acute respiratory syndrome (SARS). PMID- 12723282 TI - Global surveillance for severe acute respiratory syndrome (SARS). PMID- 12723284 TI - Current strategies in the management of spinal metastatic disease. AB - The treatment of metastatic spinal cord compression is complex. The three treatment modalities that are currently applied (in a histologically non-specific manner) are surgery, radiotherapy and the administration of steroids. The development of new spinal instrumentations and surgical approaches considerably changed the extent of therapeutic options in this field. These new surgical techniques have made it possible to resect these tumours totally, with subsequent vertebral reconstruction and spinal stabilization. In this respect, it is important to clearly identify those patients who can benefit from such an extensive surgery. We present our management algorithm to help select patients for surgery and at the same time identifying those for whom primary non-surgical therapy would be indicated. The retrospective review of surgically treated patients in our department in the last four years reveals a meagre application of conventional guidelines for the selection of the appropriate operative approach in the surgical management of these patients. The reasons for this discrepancy are discussed. PMID- 12723283 TI - [Swiss surgery: quo vadis? Reader and market analysis for strategic positioning of a specialty journal]. AB - Scientific journals currently face challenges including cost pressures caused by economic constraints, increasing rivalry among competitors, limited market potential of non-english speaking journals, increasing medical specialization with resulting market fragmentation, and internet-based competition. We therefore analyzed strategic opportunities of the journal Swiss Surgery on the basis of customer surveys and of a market analysis. RESULTS: Swiss surgeons expressed their interest in the continuation of the journal but also indicated their support for changes in its concept and for an increased use of electronic media. An international market analysis points-out the difficulties of national, non english speaking journals in gaining impact points and in attracting authors and readers of scientific medical articles. Therefore, a journal such as Swiss Surgery should identify and use publication niches. RECOMMENDATION: The demand for a concept addressing surgical training including continuous postgraduate education was confirmed by the customers of Swiss Surgery. A corresponding offer does not presently exist in the area and could become the new focus of the journal. This change of concept may have a number of consequences: A journal focusing on surgical training and education should use the results of readers' surveys rather than impact point assignment to evaluate quality. The journal should increasingly use electronic services including data bases, pictures, videos and closed user groups to supplement the print version. At short term, however, the printed version should be continued and not be substituted by the electronic version in order to maintain the established brand "Swiss Surgery". PMID- 12723285 TI - [Which lymphadenectomy in papillary thyroid gland carcinoma?]. AB - BACKGROUND: The optimal treatment of papillary thyroid carcinoma (PTC) is still debated, also with respect to nodal treatment. METHOD: Retrospective analysis of a personal series of 159 patients with PTC, with respect to nodal disease, follow up 1-27 (mean 8) years. RESULTS: In 42 patients with clinical, macroscopic nodal disease (cN1) a therapeutic lymphadenectomy was performed, with pN1 status in 41 (98%) patients. 117 patients had no clinical or intraoperative suspicion of nodal involvement (cN0), with occult nodal disease in 5/29 (17%) patients undergoing prophylactic (elective) lymphadenectomy, and in 2/88 (2.3%) patients without primary lymphadenectomy (metachronous nodal disease) (p < 0.005). Nodal recurrences were observed (1-5 years after primary treatment for cure) in 5/42 (12%) pN1 and in 3/114 (2.6%) cN0, pN0 tumors (p = 0.009), with unfavourable outcome in 2 and 1 patients, respectively. The 20-year tumor specific survival was 100% in TNM I + II (low risk) patients (including pN1 and N0 tumors); the survival rate was deteriorated by stage pN1 vs. N0 in TNM high risk patients (50% vs. 86%; p = 0.03). DISCUSSION: The intraoperative macroscopic staging (cN) remains important:--clinical nodal disease warrants a systematic node dissection (microdissection), for preventing (curable or serious) nodal recurrences. Occult nodal disease does not evolve frequently in clinical nodal recurrence. A less radical (and only central) prophylactic lymphadenectomy, avoiding surgical morbidity, may be oncologically adequate. More sensitive detection of nodal positivity (frozen section of sampling tissue or sentinel nodes, immunohistochemistry) appears not rationale. In pN0, cN0 tumors use of prophylactic 131I may represent overtreatment, and follow up controls may be conducted less rigorously. PMID- 12723286 TI - [Intramedullary Kirschner wire osteosynthesis in treatment of distal metacarpal fractures]. AB - The intramedullary Kirschner wire fixation of distal metacarpal fractures reported by Foucher et al. combines the known advantages of intramedullary implants with a reduction of iatrogenic soft tissue trauma. We applied this minimal invasive internal fixation technique in 38 patients with fractures dislocated by more than 20 degrees and/or with rotatory deformity. After opening the intramedullary cavity from the base of the respective metacarpal bone and after fracture reduction, two pre-bent Kirschner wires were intramedullary inserted in an orthograde fashion. The pre-bent distal end of the wire in the form of a hockey club allows an additional closed reduction of the displaced distal fracture fragment. Intraoperative complications did not occur. A fixation in a plaster splint followed for one week only. The elastic fixation of the wires working as springs is stable enough to allow physiotherapeutic exercises. After the third postoperative week, the intensity of physical exercise was increased. 36 of the 38 patients were followed. With one exception, all fractures were healed in a proper position. The wires were removed under local anaesthesia on an outpatient basis after six to eight weeks and full mobility of the fingers was achieved in 34 patients at that time. Complications included one redislocation and one distal wire perforation. PMID- 12723287 TI - Open or closed pinning for distal humerus fractures in children? AB - INTRODUCTION: In children, the choice between percutaneous pinning (PP) and open pinning fixation (OPF) for the surgical treatment of fractures of the distal humerus remains controversial, especially the PP method for internal humeral condylar (IHC) fractures. PATIENTS AND METHODS: Eighty fractures of the distal humerus in children were treated surgically in our hospital over a ten year period. 47% (n = 38) were supracondylar (SC), 20% (n = 16) comminuted (COM), 18% (n = 14) internal humeral condylar (IHC), and 15% (n = 12) lateral humeral condylar (LHC). We used PP, OPF and three times osteosynthesis with screws. RESULTS: In comparison to OPF, PP reduced the length of hospitalization in SC fractures (2.8 versus 6.1 days) and IHC fractures (2.4 versus five days). It reduced the risk of extension deficiency (11.1% versus 15%) and of cubitus valgus (0% versus 20%) in SC fractures, and of cubitus varus in IHC fractures (0% versus 11.1%). However it induced a higher rate of cubitus valgus (11.1% versus 20%) in IHC fractures, one persistent neurological motor deficiency (radial nerve) and four cases of transitional neurological involvement (ulnar nerve). CONCLUSIONS: PP is a good surgical method for SC and for also for IHC fractures, if performed by experienced surgeons so as to avoid neurological damage. PMID- 12723288 TI - Sub- or intertrochanteric fracture following screw fixation of an intracapsular proximal femoral fracture: true complication or technical error? AB - PURPOSE: To review, retrospectively, the possible causes of sub- or intertrochanteric fractures after screw fixation of intracapsular fractures of the proximal femur. METHODS: Eighty-four patients with an intracapsular fracture of proximal femur were operated between 1995 and 1998 by using three cannulated 6.25 mm screws. The screws were inserted in a triangular configuration, one screw in the upper part of the femoral neck and two screws in the inferior part. Between 1999 and 2001, we use two screws proximally and one screw distally. RESULTS: In the first series, two patients died within one week after operation. Sixty-four fractures healed without problems. Four patients developed an atrophic non-union; avascular necrosis of the femoral head was found in 11 patients. Three patients (3.6%) suffered a sub- and/or intertrochanteric fracture after a mean postoperative time of 30 days, in one case without obvious trauma. In all three cases surgical revision was necessary. Between 1999 and 2001 we did not observe any fracture after screwing. CONCLUSION: Two screws in the inferior part of the femoral neck create a stress riser in the subtrochanteric region, potentially inducing a fracture in the weakened bone. For internal fixation for proximal intracapsular femoral fracture only one screw must be inserted in the inferior part of neck. PMID- 12723289 TI - [Embolization of the splenic artery after splenic gunshot wound]. AB - Computed tomography (CT) of the abdomen is a reliable method for evaluation of spleen injuries and has the potential to exclude further abdominal injuries. Blunt and penetrating injuries of the spleen have to be managed immediately due to a high mortality rate. Two therapeutical options, conservative or operative, are currently available. In general, a hemodynamic stable patient, no further injuries of the abdominal organs or the skull as well as no history of abdominal surgery of the abdomen are prerequisites for a non-surgical therapy. Catheter based angiography gives the possibility to diagnose and to treat injuries of blood vessels of the spleen, which were seen on the CT scans. Gunshots are relatively rare in Europe, but the mortality of such traumas is high. The present case demonstrates a patient with a penetrating gunshot trauma of the left hemiabdomen with a bleeding injury of the spleen. Due to the stable hemodynamic conditions, absence of further injuries of the abdomen or the skull and, because of previous pancreas surgery a non-surgical therapy was chosen, consisting of proximal embolisation of the splenic artery. PMID- 12723290 TI - Spontaneous splenic rupture--a rare complication of amyloidosis. AB - We present a case of spontaneous rupture of the spleen, an uncommon complication of systemic amyloidosis. Amyloid deposition leading to capsular distension and increased vascular fragility is thought to predispose the spleen to rupture spontaneously. PMID- 12723291 TI - [Cantilever resin-bonded bridges with one adhesive surface. A review of the literature]. AB - Conventional resin-bonded bridges are designed with adhesive surfaces on both sides of the dummy. As a consequence both abutment teeth are rigidly connected to each other. Loading of the abutment teeth or dummy will lead to wringing forces in the cement. Adhesive bridges with one adhesive surface are actually cantilever bridges. Theoretically this type of bridges will be less prone to wringing forces due to loading. However, long term research into the performance of cantilever resin-bonded bridges (RBBs) is hardly available. The results of research into the longevity of conventional RBBs differ remarkably between Europe, and the United States and Japan. These difference are discussed in this article. This article compares the fixed-fixed design RBBs with the cantilever RBBs. The differences are discussed based on eight articles, the earliest being published in 1991, which are dedicated, completely or partially, to the subject of cantilever adhesive bridges. One can conclude that in The Netherlands RBBs are wrongly considered to be unreliable. On the contrary, they appear to be reliable and predictable restorations provided their preparations meet the right standards. Although a number of authors conclude that cantilever RBBs are performing better than their fixed-fixed design counterparts in similar situations, further research is needed concerning the longevity of this type of adhesive bridges. PMID- 12723292 TI - [A finite element model of a cusp-replacing adhesive restoration]. AB - This article describes the development of a three-dimensional finite element model of a premolar, based on a micro-scale computed tomographic data-acquisition technique. Using the model shrinkage stresses were analysed during and after the polymerisation process of resin composite. The stress patterns generated were three-dimensional. The results of this study indicate that failure of the interface is more probable than failure of the composite material. The described procedure is a relatively easy method to produce a highly detailed 3-D finite element model of a premolar with an adhesive cups-replacing restoration. PMID- 12723293 TI - [Cosmetic dentistry. An ethical reflection on the boundaries of embellishment]. AB - From a technical point of view the possibilities of aesthetics in medicine and dentistry are unlimited. This makes the call for moral boundaries a number one topic. It is especially this 'question of boundaries' with regard to the realisation of embellishments, which is considered problematic: who determines the moral boundaries of dental intervention for embellishment purposes and where do those moral boundaries of embellishment lie? Considering the increasing possibilities it is not advisable to just leave this to the individual dentist, let alone to the patient. Te vision of the profession is needed, so that the individual dentist feels supported. PMID- 12723294 TI - [A mistake in the picture processing procedure?]. AB - A 12-year old boy was referred to an orthodontist. During examination of his dental status a swelling was seen at the right side of his palate. Further examination demonstrated a central giant cell granuloma. This is a rare benign tumor that often causes local destruction of the bone and displacement of teeth. Surgical curettage of these large central giant cell granulomas gives mulilations of the jaws and loss of teeth and tooth germs. Furthermore, it has a high recurrence rate. Therefore, calcitonin was chosen as an alternative therapy. PMID- 12723295 TI - [The plague]. AB - The word plague evokes associations with old times, but plague actually does still occur all over the world. The survival chances are at the moment much better (antibiotics) than formerly, when half of the patients died; now this figure is 15%. The causal organism is Yersinia pestis, a bacterium in fleas on rodents. This bug is a plausible candidate for attacks by bioterrosists. So, in this time, there is good reason to pay attention to it, as well as to to smallpox and anthrax and their causal organisms. PMID- 12723296 TI - Employment-based retirement and pension plan participation: geographic differences and trends. PMID- 12723297 TI - Clarian Health focuses site upgrade on personalization and integration. PMID- 12723298 TI - Clinical trial sites only marginally effective. PMID- 12723299 TI - DM on the Web. PMID- 12723300 TI - Personalized functionality will help bring them back. PMID- 12723301 TI - Syndromic surveillance. PMID- 12723302 TI - Concerns remain as healthcare facilities embark on smallpox vaccination. PMID- 12723303 TI - [Flexor tendon pulley system: anatomy, pathology, treatment]. AB - Flexor tendon pulley has been very early noticed and described. Terminology usually accepted recognizes 6 arcifom pulleys (A0 to A5) and 3 cruciform pulleys (C1 to C3). Anatomy and physiology of this flexor tendon gliding and reflection system at the level of the digital sheet are exposed. The integrity necessity of this system became obvious regarding the flexor tendons repair. Four main pathologies may be concerned: the trigger finger congenital or progressive, due to a chondroid metaplasia of the A1 pulley; tenosynovial ganglions arising at the weak point between A1 and A2 pulley; lesions of the flexor tendon sheet during traumatic lacerations or surgical repairs; quite experimental lesions creating isolated ruptures of one or several pulleys which occur during sport practice, especially high level rock climbing. The repair techniques are exposed to allow to graduate and hierarchy the reparation technique regarding the pathology. A2 and A4 repair is always indicated. The best reconstruction material is an extensor retinaculum graft. But its poor surface available often draws to use conventional palmaris longus free graft. PMID- 12723304 TI - [Surgical treatment of chronic mallet finger by shortening--suture of the tendon scar. Sixty six cases]. AB - Acute mallet fingers are commonly treated by splinting. Treatment of chronic injuries is more debated. Since 1989, a "shortening and suture" technique have been used for such chronic injuries on the elongated tendon scar. Sixty six of 77 patients treated on a 10 years period were reviewed with a mean follow-up of 21 months. The mean active extension lag at the distal interphalangeal (DIP) joint was 4.5 degrees (41 degrees of improvement) with 52% of fingers which recovered a full extension, representing 77% of good and excellent results according to Abouna's and Brown's modified criteria. There were two failures which lead to reoperation, and no complication (2 painful scars and 20% of cold intolerance). We propose this safe and simple technique for chronic mallet fingers if deformity exceeds 30 degrees, for patients untreated (after the second month), or when splinting has failed. "Swan-neck" deformities were improved by an associated Fowler procedure. In case of failure, a new "shortening and suture" or a DIP arthrodesis can be discussed. PMID- 12723305 TI - Extensor tendon repair: mobilise or splint? AB - OBJECTIVES: The aim of this study was to assess the static and dynamic hand therapy regimes used at Mount Vernon Hospital, following extensor tendon injury during 1995-2000 and compare them to the early active regimes published. METHODS: Sixty-five patients were included and their hand function recorded by calculating total active motion (TAM), percentage combined motion and extensor lag at 4-6 weeks and at 10 weeks postinjury. RESULTS: The results in two groups were good, with mean TAMs of 202 and 258 at 4-6 weeks and at 10 weeks, respectively, for the static regime and 214 and 245, respectively, for the dynamic regime, during the same time periods. CONCLUSION: The results from each group compare favourably with the published series of patients undergoing early active motion, where mobilisation is commenced almost immediately. The authors' preference is the static regime as it is simple, effective and particularly useful in poorly compliant patients. PMID- 12723306 TI - [Extensor tendon rupture after dorsal surgery of the rheumatoid wrist: analysis of nine reviewed cases]. AB - We led a retrospective study to determine the causes of the tendon ruptures post operating in the surgery of the wrist rheumatoid dorsal and to estimate the clinical result. At follow-up, we measured the extension lag and the rolling-up of fingers by the distance palm-pulps. Nine patients were so revised in the average of 40 months, average age was of 50.7 years. The tendon ruptures arose in 3 months in 67% of the cases. Seven times, a procedure on the distal radio-ulnar joint had been necessary (5 Sauve-Kapandji and 2 Darrach). Thirty tendons had been concerned in this study, that is 3.3 tendons on average (1-5). Two main causes were found: attrition on the stub ulnaire and great intra-tendinous synovitis (per operating observation). At the revision, the lag extension means was 23 degrees (0-40). Rolling-up of the long fingers was complete 4 times on 7. The best results were observed after tendinous grafting or index proprius transfer with a lateral suture. Tendinous adhesions had arisen 6 times and persisted still at 3 patients. Our study underlines the interest to stabilize the stub ulnaire to prevent the post-operating ruptures and proposes a transfer or a graft in front of tendons very weakened by the synovitis. PMID- 12723308 TI - [Treatment of piso-triquetral pain by excision of the pisiform: report of fifteen cases]. AB - INTRODUCTION: Piso-triquetral joint could be painful and pisiformectomy can be the best procedure if conservative treatment have failed. The aim of this retrospective study was to evaluate clinical and functional results of pisiformectomy. MATERIAL AND METHOD: Thirteen patients with pisiformectomy were included in this retrospective study, 2 of them have bilateral procedure (n = 15). An ulnar neuropathy was associated in four cases. The average follow-up was 31.5 month. Professional traumatic injuries were the most common etiology. Surgical procedure was the same for all patients. Patients were reviewed clinically and subjective results were evaluated with an analogic scale (Eva). RESULTS: No postoperative complications were found. At the maximal follow-up, result was excellent for 12 cases, good for 2 cases and fair for one case. Analogic evaluation pain at maximal follow-up was 0.8 points compared to 6.4 points preoperatively (p < 0.001). Wrist range of motion was improved. DISCUSSION: Etiologies of piso-triquetral disorders were large and dominated by traumatic or microtraumatic injuries. Pisiformectomy is the best procedure if conservative treatment have failed. There is no place for piso-triquetral arthrodesis considering pisiformectomy's good results. Nevertheless, it's important to know that piso-triqueral disorder could be included in an authentic loco-regional disorder. PMID- 12723307 TI - [Guepar anatomical trapeziometacarpal prosthesis]. AB - INTRODUCTION: A choice of surgical techniques of treatment for trapeziometacarpal (TMC) Osteo-Arthritis (OA) have been described. Total arthroplasty is often used, especially in France. Many papers have been published, presenting various prostheses. In English literature, this device is not thoroughly used. MATERIALS AND METHODS: [corrected] Guepar total arthroplasty is a cemented ball-in-socket prosthesis in metal-polyethylene. It includes an anatomical stem available in 4 sizes. After failure of the conservative treatment, total arthroplasty must be reserved to elderly patients, painful, with OA Dell stage III or IV aligned or not. The trapezial height must be sufficient. The authors reports the preliminary results of 64 Guepar prostheses, anatomical new design, implanted since 1995. RESULTS: Results of 63 prostheses are presented. One removal had been necessary at 9 months for metacarpal loosening (failure). Mean follow-up was 29 months. Clinical results were judged excellent or good in all cases. Regarding the radiological results, no modifications has been observed in 56 cases. Six radiolucent lines without displacement of the implants has been noted, with no incidence on clinical results. In one case, a metacarpal stem penetrated into the medullary canal in the bone axis but without any clinical modifications. DISCUSSION AND CONCLUSION: Clinically, in addition to pain relief, trapeziometacarpal prosthesis allows to preserve the first column length and to obtain a better opposition of the thumb as well of a better thumb-digits pinch, compared after trapeziectomy. Radiologically, as for total hip arthroplasty, the exact adaptation of an anatomical stem (new design) to the canal has probably a better prognosis at long term follow-up. PMID- 12723309 TI - [Finkelstein's versus Brunelli's test in De Quervain tenosynovitis]. AB - This short paper demonstrates that the Finkelstein's test in De Quervain's tenosynovitis is based on an incorrect assumption. The correct basis for a pathognomic manoeuvre in De Quervain is the provocation of tendons attrition of the first wrist dorsal compartment against their pulley which elicits pain. The Brunelli's test induces this friction and pain by asking the patient to hardly adduct the thumb with the wrist in radial deviation. PMID- 12723310 TI - Gouty involvement of flexor tendons. AB - This article presents three patients suffering from complications related to tophi deposited within the hand and wrist synovium and flexor tendons. One patient had no previous history of gout or acute arthritis, with uricemia within normal values upon admission. The pathophysiology and treatment of gout in these special circumstances are discussed. PMID- 12723311 TI - [Extraskeletal chondroma of the hand, a rare cause of carpal tunnel syndrome]. AB - The authors report the case of a 18 years old girl presenting with a carpal tunnel syndrome, with no history of trauma and a progression of the symptoms in 2 months. Surgery was performed and the diagnosis seems to be an extraskeletal chondroma of the hand. PMID- 12723312 TI - [Orthopedic and traumatologic surgery; peripheral nerve surgery; surgery of the hand]. PMID- 12723313 TI - [Correlation of risk factors with the efficacy of applied therapies by breast conserving procedure in breast cancer]. AB - 1007 cases of female breast cancer patients treated with breast conserving surgery and subsequently irradiation with a median dose of 66 (50-80) Gy including boost with tangential high voltage photon beams. 34.6% (348/1007) received no further therapies, 53.4% (538/1007) Tamoxifen, 26% (262/1007) an adjuvant chemotherapy +/- Tamoxifen. All tumors were classified on the basis of the pathologic-anatomical spreading: 70.7% (712/1007) pT1a-c, 27.4% (276/1007) pT2. 1.9% (19/1007) pT3-4 due to the refusal of mastectomy or an error in the preoperative diagnosis. 32.5% (327/1007) showed proven axillary metastases, of which 26.3% (86/327) > or = 4 LN+. Median age 56 (23-92) years. The local relapse rate after a median follow-up of 70 (12-264) months amounted to 5.9% (59/1007). Distant metastases were registered in 11.5% (116/1007). A total of 8.8% (89/1007) died as consequence of breast cancer, 3.2% (32/1007) of other causes. In 82.6% (816/988) of the pT1/pT2 tumors the resection area had been described. In 29.8% (156/524) in the resected parts there were found rests of tumors. The LRFS falls from 94% to 82% and by remained R1 (26/524) to 47%. Correlation likewise the DMFS, which sanks from 81% to 68% respectively to 63%. We expect a second wave of metastases like the situation by local relapses. Often the R1-resection was connected with other histological high risk factors as multifocality/-centricity, necrosis or vascular invasion. If one divides the patient case sample into a first group with special risk factors (< or = 40 years of age, > or = 4 positive axillary lymph nodes, vascular invasion), and a second which exhibited none of these components, the first group had a 23-26% lower disease free survival rate. Amazing is the fact that, subsequent to a lumpectomy and irradiation, the use or non use of Tamoxifen and/or cytostatics was without proven statistical significance. The evaluation was conceived and implemented more than 20 years ago, and documentation was continuously collected ever since. We're aware of the lack of randomization, but there are less the randomized studies than rather its transformations respectively the daily routine who will decide about life and death. However, evaluations of this data by medical oncologists would, on the one hand, make it possible to better assess the importance of the available data and our results, and, on the other hand, clarify the clinical value of partially and/or completely applied medical treatments. PMID- 12723314 TI - [Inclusion body myositis (IBM) -- a review]. AB - Inclusion body myositis (IBM) is characterized by the insidious onset of slowly progressive proximal and distal weakness. The clinical hallmark of IBM are atrophy and weakness of the quadriceps and the wrist and finger flexors. Although frequently misdiagnosed, IBM is the most common inflammatory myopathy in patients over the age of 50 years. The diagnosis of IBM can be made, even in absence of a typical clinical history, exclusive on the basis of muscle biopsy when all of the characteristic histopathological findings are present (inflammation, rimmed vacuoles, protein deposits, and 15-18 nm tubofilaments). Apart from sporadic IBM there is a group of heterogeneous inherited myopathies with histopathologic similarities to sporadic IBM. Although some immunomodulating therapies may exert transient and mild benefits, there is no effective treatment for IBM. PMID- 12723315 TI - [Delirium. A 74-year-old retired butcher's shop employee]. PMID- 12723316 TI - [Teenager with fatigue and hepatosplenomegaly. A 15-year-old secondary school student]. PMID- 12723317 TI - What Joint Commission's alert on infections means for ORs. PMID- 12723318 TI - Implant prices see largest jump since 1992. PMID- 12723319 TI - Top hospitals have better outcomes, lower costs. PMID- 12723320 TI - Top 100 hospitals. Hospital's culture values employees. PMID- 12723321 TI - Patient privacy and implant tracking. PMID- 12723322 TI - How is block time reviewed and revised? Part 3. PMID- 12723324 TI - Top ten safety issues with medical devices. PMID- 12723323 TI - AAMI gives nod to new steam indicators. PMID- 12723326 TI - What's best way to market your ASC? PMID- 12723325 TI - Taking steps to control patients' pain. PMID- 12723327 TI - Some thoughts on patient compliance. PMID- 12723328 TI - Orthodontic recycling at the crossroads. PMID- 12723329 TI - My life as a locum tenens. PMID- 12723330 TI - Herbst appliance variations. PMID- 12723331 TI - The hybrid core system for indirect bonding. PMID- 12723332 TI - Clinical management of unilaterally impacted mandibular first and second molars. PMID- 12723333 TI - Evaluating and choosing a fund-raising database program. PMID- 12723335 TI - 2003 non-profit software guide. PMID- 12723334 TI - American Red Cross releases September 11 report. PMID- 12723336 TI - HIPAA. Health Insurance Portability and Accountability Act of 1996. AB - Organizations must obtain a copy of the final privacy rule. For those organizations that have acted in advance and developed policies it will be necessary for them to assess and revise these policies as indicated. Develop and post a notice of the organizations privacy practices and patient's privacy rights. Develop and implement a mechanism in which patients would be asked to sign an acknowledgment of receipt of the privacy notice, which would become part of the patient's treatment record. Offices will be required to provide evidence that staff has been trained on the organization's privacy policies procedures, patient's privacy rights and the consequence of violating established policies and procedures. This training should become part of the employee initial orientation. Develop a log to be kept in each patient's record to document release of information. This includes any element of the patient's medical record released to a third party (sexual abuse, substance abuse, health issues, etc.). This document should be reviewed with legal counsel. Develop a log to be kept in each patient's record to document patient authorization to leave messages on personal answering devices By ensuring consistency throughout the industry, these national standards will make it easier for health plans, doctors, dentists and hospitals to process claims and other electronic transactions efficiently and effectively. PMID- 12723338 TI - The missing link. PMID- 12723337 TI - Security in dental office computing. AB - Dental office computers are increasingly used to store critical financial and clinical data. That data is subject to loss or alteration from a number of possible sources. Secure communications, security of the computer system from outside attacks, and data back-up are discussed. PMID- 12723339 TI - Helping the beat go on! PMID- 12723340 TI - The army goes rolling along.... mobilization operations offer great experience for Fort Stewart dental assistants. PMID- 12723341 TI - [Fluctuations of pass band coefficients of water and water salt solutions in the spectral infrared region]. AB - The effect of superlow concentrations of KCl and CaCl2 solutions, obtained by diluting starting 1 M solutions of these salts 10(9)-10(15) times, on the fluctuations of pass band coefficients of water was studied. It was found that these solutions are substantially distinguished from bidistilled water by a high intensity of fluctuations of spectra in the infrared region. It was also shown that higher concentrations of solutions of these salts (at dilutions less than 10(8) times) have no strong and specific effect on fluctuations of the infrared spectrum: the intensity of fluctuations of the infrared spectra of these solutions practically coincides with that of control water. Possible reasons for this effect are discussed. PMID- 12723342 TI - [Study of kinetic parameters of singlet molecular oxygen in aqueous porphyrin solutions. Effect of detergents and the quencher sodium azide]. AB - The kinetic parameters of porphyrin-photosensitized formation and deactivation of singlet molecular oxygen (1O2) and their dependence on the concentration of the 1O2 quencher sodium azide were investigated in air-saturated water, ethanol, and aqueous micellar solutions of detergents using time-resolved measurements of oxygen phosphorescence under pulsed laser excitation. The lifetimes of 1O2 formation and deactivation and the rate constants of 1O2 quenching by sodium azide were determined. It was shown that, with no azide in the solutions, the rise in phosphorescence intensity after the laser flash corresponded to the kinetics of energy transfer from the porphyrin triplet molecules to oxygen, while the decay kinetics corresponded to the kinetics of 1O2 deactivation. In the presence of detergent, a considerable increase in the 1O2 lifetime was observed, which is likely due to the localization of 1O2 molecules mostly in lipophilic micelles and not in the water phase. If relatively high azide concentrations were used, the lifetime of the porphyrin triplet state did not change but the 1O2 lifetime decreased to values similar to those in living cells. In this case, the inversion of the phosphorescence kinetic phases was observed. The rise corresponded to 1O2 deactivation, and the decay, to the energy transfer from triplet porphyrin to oxygen. The data suggest that, in living cells, 1O2 molecules are also located mainly in lipophilic structures and the 1O2 lifetime determines the kinetics of the phosphorescence rise after the laser pulse. PMID- 12723343 TI - [Effect of disulfide bonds on lysozyme dynamics]. AB - The effect of destruction of disulfide bonds on the dynamics of proteins was studied by an example of lysozyme by the methods of molecular dynamics. In lysozyme, in the absence of disulfide bonds, the characteristic times of motions of secondary structure devices increased 3-7 times, whereas the amplitudes of fluctuations of secondary structure devices practically did not vary. In the absence of S-S-bonds, the volume of the molecule decreased approximately by 2%, primarily due to a "cleft" between the major and the small domains of lysozyme. Thus, disulfide bonds not only "glue" the secondary structure devices of the protein but also play a role of "rods", maintaining a certain free volume of the molecule necessary for the realization of its functions. PMID- 12723344 TI - [Destruction of amyloid beta-protein by exposure to weak magnetic fields]. AB - It was found that weak magnetic fields (constant 42 microT; variable 0.05 microT; summation of frequencies in the range of 3.58-4.88 Hz) substantially (approximately fourfold) accelerate the hydrolysis of the amyloid beta-protein in aqueous solution. A region of the amyloid beta-protein most sensitive to the action of weak magnetic fields was determined. It is localized between Asp and Ser amino acid residues in positions 7 and 8 of the peptide sequence. It is in this region that the hydrolysis of the molecule of the amyloid beta-protein by the action of magnetic fields predominantly takes place. PMID- 12723345 TI - [Calculation of spectra of absorption and circular dichroism of Rhodopseudomonas acidophila light harvesting complexes based on roentgen structural data]. AB - Absorption and circular dichroism spectra of two forms of the peripheral light harvesting complex from photosynthetic purple bacteria Rhodopseudomonas acidophila were calculated. Calculations were carried out on the basis of exciton theory for circular aggregates of bacteriochlorophyll molecules and X-ray data for these forms of the complex. It was shown that theoretical spectra fit well experimental ones at the same values of excitation energy, homogeneous and inhomogeneous broadening, and bandwidth for all bacteriochlorophyll molecules of complexes. To approximate the circular dichroism spectra of complexes, it was necessary to change the orientations and the values of the moments of transition of Qy molecules relative to their orientation determined on the basis of X-ray structure analysis data. PMID- 12723346 TI - [Pigment accumulation and functional activity of chloroplasts in common Pisum sativum L. mutants with low chlorophyll level (chlorotica)]. AB - Pea mutants chlorotica 2004 and 2014 with a low content of chlorophyll were studied. The mutant 2004 has light green leaves and stem, and the mutant 2014 has yellow green leaves and stem. They accumulate approximately 80 and 50% chlorophylls of the parent form of pea Torsdag cv. The content of carotene in carotenoids of the mutant 2004 was much lower, and the accumulation of lutein and violaxanthine was increased. The accumulation of all carotenoids in the mutant 2014 decreased almost proportionally to a decrease in the chlorophyll content. The rate of CO2 evolution in mutant chlorotica 2004 and 2014 was established to be lower. The quantum efficiency of photosynthesis in the mutants was 29-30% lower as compared to the control, and in hybrid plants it was 1.5-2-fold higher. It is assumed that the increase in the activity of the night-time respiration in gas exchange of chlorotica mutants and the drop of photosynthesis lead to a decrease in biomass increment. The results obtained allow us to conclude that the mutation of chlorotica 2004 and 2014 affects the genes controlling the formation and functioning of different components of the photosynthetic apparatus. PMID- 12723347 TI - [The role of lipid-nucleic acid interactions in the reconstruction in vitro of the nuclear shell with pores]. AB - Previous studies of triple complexes DNA-phosphatidylcholine liposomes-Me2+ showed that, upon complexing, along with aggregation, a partial reversible fusion of liposomes takes place. In this case, DNA acts as a fusogen. The addition of the extract from Xenopus laevis or Drosophila melanogaster oocytes to the triple complex leads to a complete fusion of initial liposomes 100-200 nm in size and the formation of giant liposomes of up to 100-500 microns in size. Upon the addition of liposomes to the extract from Xenopus or Drosophila eggs, either fragments of the nuclear envelope with pores or structures like annulate lamellae are formed. The diameter of pores in these formations is approximately twice as small as in the native nuclear envelope, which may be associated with a lower surface tension of the lipid bilayer. This fact is in agreement with our model of the formation of nuclear pores suggested earlier. PMID- 12723348 TI - [Electric capacitance of bilayer lipid membranes from hydrogenated egg lecithin during phase transition from liquid crystalline state to gel]. AB - The electrical capacity of planar bilayer lipid membranes (BLM) from natural hydrogenated egg lecithin (HEL) in n-decane at a temperature of phase transition was measured. The temperature of phase transition was determined calorimetrically to be 51 degrees C. The data obtained revealed a phase separation of HEL in BLM into two fractions, one freezing at 42-44 degrees C and one that is converted to a liquid-crystal state at 51-59 degrees C. It was assumed that the first fraction is rich in dipalmitoyl lecithin, and the second fraction is rich in distearoyl lecithin. Freezing and the transition to the liquid-crystal state were accompanied by an increase and decrease in membrane thickness, respectively, in part due to a displacement of the solvent from the torus to the planar part of the bilayer. The displacement of the solvent is explained by changes in the disjoining pressure in BLM, which arises across the lipid bilayer due to van der Waals forces of attraction between water layers on both sides of the BLM. PMID- 12723349 TI - [Changes in the lipid physical state in human erythrocyte membranes subjected lead exposure in vitro]. AB - The effect of lead acetate on the physical state of membrane lipids in human erythrocytes in vitro was studied using the lipophilic fluorescence probe 1,6 diphenyl-1,3,5-hexatriene and spin probes 16-doxyl-stearate and iminoxyl palmitic acid. It was shown that 2-10 microM lead acetate causes an increase in both intensity and polarization of fluorescence of 1,6-diphenyl-1,3,5-hexatriene, indicating changes in the microviscosity of the lipid bilayer of erythrocyte membranes. Judging from the parameters of EPR spectra of 16-doxyl stearate and iminoxyl palmitic acid incorporated into erythrocyte membranes, 2-10 microM lead acetate increases the heterogeneity of the lipid bilayer in surface and deep hydrophobic layers of the erythrocyte membrane. PMID- 12723351 TI - [Effect of diamide on deformability and La(3+)-induced aggregation and fusion of human erythrocytes]. AB - The effect of diamide on the deformability and La(3+)-induced aggregation and fusion of human erythrocytes was studied. It was shown that diamide decreased the deformability of erythrocytes and practically completely inhibited their fusion. It was found that diamide did not change the aggregation of erythrocytes. PMID- 12723350 TI - [Photohemolysis sensitized by deuteroporphyrin-IX derivatives: determination of binding strength of dyes to erythrocytes]. AB - A comparative analysis of the ability of 4-(1-methyl-3-hydroxybutyl) deuteroporphyrin-IX (I) and 2,4-di-(1-methyl-3-hydroxybutyl)-deuteroporphyrin-IX (II) to photosensitize hemolysis of human erythrocytes was performed. The photohemolytic efficiency of dye I was shown to be about 60 times higher than that of dye II. It was found that a part of each dye tightly binds to erythrocyte membranes and is not removed by washing. A method for estimating the share of the dye tightly bound to the membrane (beta) was proposed, which takes into account the shielding effect produced by the free dye and the photohemolytic efficiency of the bound dye. It was shown that the beta values for dyes I and II are 86 and 61% and correlate with the coefficients of distribution of the dyes in the octanol/water system (20.7 and 17.0, respectively). PMID- 12723353 TI - [Spontaneous mutational background of microorganisms in the absence of substrate]. AB - The dynamics of spontaneous mutagenesis in a population of heterotrophic microorganisms during incubation in the absence of organic substrate was studied. A model of this dynamics was proposed. which is determined by the excitation by an external electromagnetic field of Langmuir proton oscillations activating the tautomeric transitions of nucleotides of DNA. It was shown that the model fits well the experimental data. PMID- 12723354 TI - [Kinetic analysis of behavior of chemotactic bacterial population at the interface of two media]. AB - The behavior of a population of bacteria at the boundary with a chemotactically active water medium containing the sites of specific binding to cell receptors was analyzed. A kinetic model of chemotaxis was used for the analysis. Differences in the behavior of strains metabolizing and not metabolizing the substrate were revealed. Six phases of interface taxis were distinguished and characterized. The results of the analysis were confirmed by the densitometric data. PMID- 12723352 TI - [Reactive oxygen forms and luminescence of intact microspore cells]. AB - The participation of reactive oxygen species (ROS) in luminescence (chemiluminescence and autofluorescence induced by ultraviolet light of 360-380 nm) was analyzed. Microspores, the pollen (male gametophyte) of Hippeastrum hybridum, Philadelphus grandiflorus, and Betula verrucosa and vegetative microspores of the spore-breeding plant Equisetum arvense served as models. It was found that the addition of the chemiluminescent probe lucigenin, which luminesces in the presence of superoxide anionradicals, leads to intensive chemiluminescence of microspores. No emission was observed in the absence of lucigenin and in the presence of the dye luminol as a chemiluminescent probe. The emission decreased significantly if superoxide dismutase, an enzyme of the superoxide anionradical dismutation during which this radical disappeared, was added before the dye addition. The autofluorescence intensity of microspores decreased in the presence of both superoxide dismutase and peroxidase, an enzyme destroying hydrogen peroxide and organic peroxides. The most significant effect was noted after the addition of peroxidase, which indicates a greater contribution of peroxides to this type of emission. The fumigation with ozone, which increases the amount of ROS on the cell surface, enhanced the intensity of the chemiluminescence of microspores with lucigenin, but decreased the intensity of the autofluorescence of microspores. Exogenous peroxides (hydrogen peroxide and tert-butylhydroperoxide) stimulated the autofluorescence of pollen and vegetative spores in a concentration-dependent manner. It was shown that the formation of ROS contributes to the luminescence of plant microspores, which reflects their functional state. PMID- 12723355 TI - [Immunocorrective effect of low intensity radiation of ultrahigh frequency on carcinogenesis in mice]. AB - The effect of low-intensity centimeter electromagnetic waves (8.15-18 GHz, 1 microW/cm2, 1.5 h daily, 20 days) on the production of tumor necrosis factor, intreleukin-2, and interleukin-3 and the expression of the heat shock protein 72 in healthy and tumor-bearing mice was measured. A significant increase in tumor necrosis factor production and a slight reduction of interleukin-2 concentration were observed after exposure to microwaves; we consider these effects as adaptive response. The interleukin-3 production in healthy mice was not affected by microwaves. Low-intensity centimeter waves induced antitumoral resistance in tumor-bearing mice. Thus, exposure of tumor-bearing mice led to a significant rise in the tumor necrosis factor production and the normalization of both interleukin-2 and interleukin-3 concentration. We assume that the significant immunomodulating effect of low-density centimeter microwaves can be used for immunocorrection and suppression of tumor growth. PMID- 12723356 TI - [Three-dimentional organization of synapses and astroglia in the hippocampus of rats and ground squirrels: new structural and functional paradigms of the synapse function]. AB - The literature data and our own data on the synaptic plasticity and remodeling of synaptic organelles in the central nervous system are reviewed. Modern techniques of laser scanning confocal microscopy and serial thin sectioning for in vivo and in vitro studies of dendritic spines, including the relationship between morphological changes and the efficacy of synaptic transmission, are discussed using, in particular, a model of long-term potentiation. The organization of dendritic spines and postsynaptic densities of different categories as well as the role of filopodia in spine genesis were analyzed. It was shown that the method of serial ultrathin sections is the most effective for unbiased quantitative stereological analysis and 3D reconstructions. By using the refined method of serial ultrathin sections with subsequent three-dimensional reconstructions, the presence of giant mitochondria in hippocampal neuronal dendrites was demonstrated. It was shown that smooth endoplasmic reticulum forms a unified continuum with the outer membrane of the mitochondrial envelope within dendrites. It was suggested that this continuum provides calcium tunneling, which makes possible intracellular signal transduction during synaptic transmission. Evidence is presented indicating the presence of gap junctions ("electrical synapses") in the synapses of mammalian brain, as well as between glial processes, and between glial cells and neurons. Our data and the data of other authors show that glial cell processes form a structural and functional glial network, which modulates the functioning of the neuronal network. The connection of dendritic spines with the glial network is shown on 3D reconstructions by analyzing the neuropil volume in CA1 hippocampal area of ground squirrels in three functional states: normothermia, provoked arousal, and hibernation when brain temperature falls below 6 degrees C. The own data of the authors are discussed indicating the formation of more than five presynaptic boutons (multiple synapses) on both CA1 mushroom-like dendritic spines and CA3 thorny excrescences. On the basis of the analysis, new ideas of the organization and functioning of synapses were suggested. PMID- 12723357 TI - [Determination of glucose diffusion coefficient in the human eye sclera]. AB - The diffusion coefficients of aqueous glucose solutions in human sclera in vitro were estimated. The method is based on measurements of time-dependence of collimated transmittance of scleral samples under the action of biocompatible immersion liquids. It was shown that changes in collimated transmittance are connected with the matching between the indices of refraction of scleral scatterers and interstitial fluid. The dynamics of interstitial fluid replacement was registered by recording successively the collimated transmittance spectra in the range of 400-800 nm. For a quantitative description of the replacement process, a diffusion model was proposed, which assumes that the diffusion coefficient is constant throughout the volume of the scleral sample. Experimental results for glucose solutions of concentrations 0.18, 0.3, and 0.4 g/ml are presented. The diffusion coefficients were estimated by approximating the experimental data in the framework of the proposed model. PMID- 12723358 TI - [The study of autowave mechanisms of electrocardiogram variability during high frequency arrhythmias: mathematical modeling data]. AB - High-frequency cardiac arrhythmias are very dangerous, as they quite often lead to sudden death. These high-frequency arrhythmias are frequently produced by rotating autowaves. In the given work, the dynamics of a rotating three dimensional excitation scroll wave and the influence of this dynamics on the variability of model electrocardiograms (ECGs) were simulated with the use of the Aliev--Panfilov model for both homogeneous and heterogeneous excitable media. Model ECGs were obtained by summing up local membrane potentials, while ECG variability was estimated numerically through the normalized variability analysis. In the homogeneous medium, the stability of the scroll wave to its filament perturbations was shown to be dependent both on the excitability of the medium and tension of the filament, while in the heterogeneous medium, the scroll was shown to be unstable. It was shown that the scroll wave dynamics affects essentially the variability of the model ECGs, and the ECG variability increases as the excitation threshold value grows. It was found that, at some parameters of the excitable medium, the variability of ECGs in the homogeneous medium is higher than in the heterogeneous medium. PMID- 12723359 TI - [Interpretation of mechanical impedance of biological tissues in the three level model with the power source of vibrations]. AB - Theoretical expressions were derived for the mechanical impedance of a round piston on the surface of a viscoelastic triple layer with sublayers linked with each other and with the rigid base. The expressions were obtained on the assumption that the piston creates an even oscillatory pressure and does not create shear stresses. The calculations in the developed model were compared with corresponding calculations in the known strict model and with experimental values obtained on a relaxed and stressed human biceps. The results obtained suggest that models of this kind can be used for reconstructing the mechanical parameters of multilayer biological tissues. PMID- 12723360 TI - [Frequency discrimination by the bottle-nosed dolphin and the Northern fur seal depending on sound parameters and sound conduction pathways]. AB - Underwater differential frequency hearing thresholds in the Black Sea bottle nosed dolphin (Tursiops truncatus p.) and the northern fur seal (Callorhinus ursinus) were measured depending on signal frequency and sound conduction pathways. The measurements were performed by the method of instrumental conditioned reflexes with food reinforcement under conditions of full and partial (with heads out of water at sound conduction through body tissues) submergence of animals into water. It was shown that in a frequency range of 5-100 kHz, underwater differential frequency hearing thresholds of the bottle-nosed dolphin changed from 0.46-0.60% to 0.21-0.34% and depended little on sound conduction pathways. The minimum underwater differential frequency hearing thresholds of the northern fur seal corresponded to the frequencies of maximum hearing sensitivity, changed from 1.7% to 1-2.3% in a frequency range of 1-20 kHz, sharply increased at the edges of the frequency hearing perception range, and depended little (in a range of 5-40 kHz) on sound conduction pathways. Thus, underwater sounds propagating through the body tissues of dolphin and fur seal reach the inner ear. PMID- 12723361 TI - [Modeling of the forest insect population size: a game theory approach]. AB - A game theory model of insect population dynamics is proposed. For the case when the population may be in one of two states: when physiological processes are directed to growth and reproduction, and when physiological processes are directed to the development of defense reactions, outbreaks of mass reproduction of insect populations may occur in conditions when population and environment have the "memory", and the state of population and environment depends on their state at the previous time moment. In the framework of the model, the well known effect of insect phase variation during the outbreak of reproduction is explained. PMID- 12723363 TI - [Generation of valuable information and the problem of goal self-setting in living systems]. AB - The problem of origination of capacity for goal self-setting is discussed. It was shown that the definition "goal" in living systems differs from the definition "target function" in physical problems concerned with nonliving systems. It was also shown that the main goal of the elements of a system is the storage of information. In biology, this goal is the extension of the principle of struggle for existence. Conditions were determined that the dynamic system describing the goal self-setting process must satisfy. It was shown that living systems meet these conditions. In inorganic nature, such systems may also arise but only as a result of long-term evolution, after which they become living. PMID- 12723362 TI - [Population size and areas of stratification in the variational model of ecosystem]. AB - The formulations of the variational task for finding the relative population size of species in community at a stationary stage of growth and of the theorem of stratification are given. Algorithms of finding the relative size of populations for communities consisting of two and three species consuming two or three resources were obtained. The borders of areas of stratification were described in which one, two, or three resources are limiting. For two species and two resources, the formulae of the dependence of relative size on the ratio of resources were derived, and the shape of this dependence for real requirements of species was demonstrated. PMID- 12723364 TI - [The study of the mixing layer by the point image method]. AB - The properties of the mixing layer in dynamic systems were studied by the example of a mathematical model of the cubic image type. Its role in the generation of information and the evolution of its significance was shown. At the moment of generation, information is of zero significance, and this significance then increases. A criterion of efficiency was proposed, and the optimum moment of making a decision in creative work was determined. It was shown that the increase in the variability of the parameters of a living system upon entry into the mixing layer and its decrease upon exit can serve as objective indicators of the transition of the system from one dynamic regime (attractor) to another. PMID- 12723365 TI - [The scaling method of spatial distribution of keratinocytes in the basal layer of epidermis in norm and during the development of psoriatic lesion]. AB - On the basis of the modern conceptions postulating that the growing layer of normal epidermis contains two populations of keratinocytes differing greatly in the rate of mitotic division (stem and transient cells), a hypothesis was proposed that explains the reason for the establishment of the portion of the proliferating fraction close to 60%. A relationship between this value and the distribution of keratinocytes of the basal layer in space was demonstrated, and formulae for its calculation were derived. Possible pathways of the transition from the normal spatial configuration to the distribution typical for the focal lesion in psoriasis were analyzed when all stem cells begin to divide vigorously, and the size of the population of transient cells is 100%. PMID- 12723366 TI - [The study of effects of geomagnetic disturbances at high latitudes on the intrauterine condition of fetus by cardiotocography]. AB - The effect of geomagnetic disturbances on the intrauterine condition of the fetus in high latitudes was studied by comparing the index of fetus condition (IFC) with the heliogeophysical indices (planetary Kp index and Dst variation index). The functional state of fetus at late terms of pregnancy was assessed by the method of cardiomonitoring. The data of fetal cardiomonitoring were processed by the method of Prof. Demidov by calculating the integral IFC using a computer program. It was found that the variations in maximum IFC values (approximately 15% of the total number of cases) coincide with seasonal variations in geomagnetic disturbances. The maximum values of IFC were observed at the equinox (March-April-May, October-November), and the minimum values were registered in the period of the solstice. A comparison of IFC values with the indices of geomagnetic disturbances indicated that the IFC value was maximum in the periods of either the greatest weakening of geomagnetic activity or maximum disturbances. PMID- 12723367 TI - [Response of the human body to heliogeophysical disturbanes]. AB - A correlation between variations in the medical parameters and the heliogeophysical indices describing physical processes during solar storms was established. It was proposed that one of the reasons for the biotropic effects observed is rheological shifts of human blood during enhancement of solar activity. The influence of heliogeophysical disturbances on the rheological properties of blood was investigated. PMID- 12723368 TI - [Essentialism and typological thinking in biological systematics]. AB - In biological literature, essentialism and typological thinking are believed to be incompatible with evolutionary ideas. At present, the same considerations underlay the claims to abandon the Linnaean hierarchy, or the fundamental classificatory structure rooted in essentialism. This paper suggests to reconsider the negative views of Plato's typology and Aristotle's essentialism following the narrow interpretations that have nothing to do with the classification of living beings. Plato's theory of 'ideas' (or 'forms') is the basis of classificatory theory; it provided such concepts as 'species', 'genus', 'essence', 'dichotomous division' but the development of this theory in the framework of moral and esthetic values could not be beneficial to biology. Aristotle's essentialism is more complicated and exists in two forms; one of these, or classificatory essentialism, is a modification of Plato's typology; another one, or organismal essentialism, represents the shift of 'essence' from the world of relations between objects to the realm of particular things, where the concept of essence lost its basic meaning. It is senseless to look for unreal 'type of an organism' ('essence of a thing') but precisely this kind of essentialism is attractive for biologists and philosophers. Organismal essentialism is the underlying basis of so-called 'individuality thesis' that is used as a weapon against classificatory essentialism. The same thesis is associated with an extensional vision of taxa that also explains the criticism of Linnaean hierarchy, while the latter is the intentional structure and the first tool suggested for the rank coordination of many unequal taxa. PMID- 12723369 TI - [The regularities in connection between fecundity with body weight and growth rate in fishes]. AB - The absolute and relative fecundity of freshwater, anadromous and marine fishes (102 species and subspecies from 33 families) and its dependence on body mass and growth rate were analyzed on the basis of published data. According to the spawning type all studied fishes were divided into species with short-term and single spawning and fishes with extended or long-term spawning. The equations of dependence of absolute fecundity (E) on body mass (W) were calculated: E = 1.033 W0.578 (the first group) and E = 0.792 W0.74 (the second group). If W < 177 g the equations don't differ significantly and one may use the equation E = 1.34( 0.742) for both groups. The body mass of females at age of maturity expressed as a portion of maximal definitive body mass equals 0.22 +/- 0.044 for many different species with 0.95 probability. The relative fecundity (a1) of some species negatively correlates with maximal body mass of adult individuals (Wmax). This dependence is expressed by equations: a1 = 3.033 Wmax-0.549 (for the first group) and a1 = 1.726 Wmax-0.351 (for the second group). Value of ratio Wov/W of different fish species changes irregularly from 0.054 to 0.32 and its average is 0.150 +/- 0.012 for the first group and 0.156 +/- 0.007 for the second one. In such a way, single reproduction effort of fishes is approximately 0.15. Comparison of data on Pisces, Crustacea, Amphibia, Reptilia, and Mammalia revealed that reproduction effort of different aquatic and terrestrial invertebrates and vertebrates varies within rather narrow limits (from 0.05 to 0.44). Average values of this index varies even less--from 0.097 to 0.238, on average 0.162, i.e. approximately 15-18% of animals' body mass falls on their reproduction constituent. PMID- 12723371 TI - [Possible ways of negative influence of acid gases on plants]. AB - The degree of negative influence of acid gases on plants is considered in dependence of their solubility in water. The linkage of water in the processes of hydration of gases forming acids can decrease the chemical potential of water in leaf apoplast. It causes the decrease in water inflow into leaf symplast. The more solubility of acid gases in water and the higher their concentration in the air, the lower water inflow from apoplast to symplast. At high concentration of toxicant water chemical potential in leaf apoplast is lower, than in symplast, and the water flows out into apoplast, i.e. plasmolis takes place. Plasmolis leads to the increase in toxicant concentration in leaf symplast and finally to the necrosis of cells. When air with acid gases are dissolving in apoplast water, "concentrating" of acid gases takes place because the acid components are more soluble in water than the main components of the air (nitrogen and oxygen). The lower acid dissociation in apoplast water, the higher speed of receipt from apoplast to symplast and even to inner cell compartments through cell membranes. It can explain why sulfur dioxide and fluoric hydrogen forming weak acids, are more toxic than nitric dioxide. Exogenous acids producing the hydrogen ions negatively influence on different metabolic processes of plants. PMID- 12723372 TI - [The possible contribution of late pleistocene biota to biodiversity in present permafrost zone]. AB - During the last decade a wide range of biological objects, which have preserved their viability for tens and hundreds of thousands of years, was found in the samples of permafrost sediments from North-East Eurasia. Among them are bacteria, fungi, algae, moss spores, seeds of higher plants, protists. Along with physiological mechanisms of cryoconservation and constant low temperature of great importance for long-term preservation of biological objects in permafrost layers are ways of burying the organisms and conditions that prevail before the transition of sediments to the permafrost state. The analysis of viability showed by preserved biological objects gives reasons to suppose that some representatives of Pleistocene biota buried in permafrost thickness may contribute to the biodiversity of present cryolite zone. PMID- 12723370 TI - [The budget model of ecosystem of a shallow highly eutrophic lake]. AB - A model of energy budget of Lake Bolshoi Okunenok ecosystem was based on the data received during field studies from May through November 1986. The model takes into account 36 components including dissolved organic matter, bacteria, phytoplankton, zooplankton, meiobenthos, macrobenthos, fish, suspended and sediment detritus. The growing season has been divided into 16 intervals according to the number of observations. The balance equation for each live component describes the change in its biomass for a time interval between two successive sampling dates. The change is considered as a balance of energy input with assimilation or feeding, and energy loss due to respiration, excretion, predation, natural mortality, fishery catchment or and emergence of imago insects. For non-live components we estimate an increase and a decrease in their mass due to the activity of living organisms, as well as organic matter exchange between water and sediments. Seasonal value of balance elements for each component are equal to sums of appropriate interval value. Comparison of energy flows through different links of a trophic web has shown that the role of a bacterial-detrial link was extremely important in Lake Bolshoi Okunenok for the growth season of 1986. Detritus constituted 58% of seasonal diet of non-predatory zooplankton, 39% of diet of predatory zooplankton, 50% of diet of planktivorous fish (fry of whitefish) and 92% of diet of benthivorous fish (fry of carp). The contribution of bacteria to the total seasonal decomposition amounted to 46%. Approximately 57% of the forage phytoplankton production, 86% of non-predatory benthos production, and 23-38% of the other trophic groups production were consumed by all grazers. "Coefficient of energy transformation" is proposed. It is calculated as: CET(s, k) = Ps(k)/Pk, where Ps(k) is production of consumers "s", built due to consumption of source "k"; Pk is production of source "k" itself. In Lake Bolshoi Okunenok only 14% of energy built by phytoplankton were accumulated in organic matter of zooplankton due to direct consumption. PMID- 12723374 TI - [On interrelation between genetic systematics and genomics]. AB - Review papers describing recent achievements of genomics usually do not pay attention to direct interrelation between genomics and genosystematics (DNA systematics). Genomics on general is based in complete DNA sequencing of genomes. Initial aim of genosystematics was the same. Absence of historical perspective in review papers devoted to genomics decreases its value. In case it is done deliberately it becomes the problem of scientific ethics. It is postulated that genomics is a natural stage of genosystematics (DNA-systematics) development. Russian scientists were among the founders of these branches of biology. PMID- 12723373 TI - [Plants in urbanized natural environment: florogenesis, coenogenesis and population structure]. AB - Published data on both flora and phytocoenosis genesis and plant population dynamics are reviewed. Urbanized environment is rated as a stress factor for single plants and their communities. The change of plant population structure in cities and towns is analysed as the important indicator of the environment quality. The problems of phytoindication of environment quality in cities and towns for its optimization are considered. PMID- 12723375 TI - [Hospital malnutrition]. PMID- 12723376 TI - Pharmacological nutrition in inflammatory bowel diseases. AB - Inflammatory Bowel Diseases--ulcerative colitis and Crohn's disease--are chronic gastrointestinal inflammatory diseases of unknown etiology. Decreased oral intake, malabsorption, accelerated nutrient losses, increased requirements, and drug-nutrient interactions cause nutritional and functional deficiencies that require proper correction by nutritional therapy. The goals of the different forms of nutritional therapy are to correct nutritional disturbances and to modulate inflammatory response, thus influencing disease activity. Nutritional intervention may improve outcome in certain individuals; however, because of the costs and complications of such therapy, careful selection is warranted. Total parenteral nutrition has been used to correct and prevent nutritional disturbances and to promote bowel rest during active disease, mainly in cases of digestive fistulae with a high output. Its use should be reserved for patients who cannot tolerate enteral nutrition. Enteral nutrition is effective in inducing clinical remission of disease in adults and promoting growth in children. Recent research has focused on the use of specific nutrients as primary treatment agents. Although some reports have indicated that glutamine, short-chain fatty acids, antioxidants and immunonutrition with omega-3 fatty acids are an important therapeutic alternative in the management of inflammatory bowel diseases, the beneficial reported effects have yet to be translated into the clinical practice. The real efficacy of these nutrients still need further evaluation through prospective and randomized trials. PMID- 12723377 TI - [Local reference intervals for the excretion of creatinine in urine for an adult population]. AB - OBJECTIVE: To present the height- and sex-specific reference intervals for the urinary excretion of creatinine in cuban subjects with ages between 19-58 years. BACKGROUND: The standards advanced by Bistrian (1985) for anglosaxon subjects have been traditionally used for the construction of the Index of Creatinine Excretion (ICE). This practice could lead to diagnostic errors in the case of ICE calculation for non-anglosaxon populations differing in dietetic behaviour and body composition. METHODS: Values of urinary excretion of Creatinine measured in 103 men and 112 women with ages between 19 and 58 years, and preserved nutritional wellbeing (weight within the interquartile range for height; 18.5 < or = body mass index < or = 30.0 kg/m2) and renal function (serum Creatinine < or = 132 mumol/l) were drawn retrospectively from the databases of the Section of Urinalysis, Service of Clinical Laboratory, "Hermanos Ameijeiras" Hospital (La Habana, Cuba). For each sex, the regression line urinary-Excrecion-of-Creatinine = c + d. height, where c, d: parameters dependent on the subject's height, were constructed. The predicted values of the urinary excretion of Creatinine for the cuban subjects were derived from the 100 (1-alpha)% prediction intervals (PI) of the regression line. RESULTS: The urinary excretion of Creatinine for a 170 cm height, 66.5 kg-weight, cuban male with age between 19-58 years, was 1216.19 mg/24 hours (95% Pl: 821.02-1611.36 mg/24 hours). The urinary excretion of Creatinine for a 160 cm-height, 59.7k Kg-weight, cuban female with age between 19 58 years, was 852.00 mg/24 hours (95% Pl: 420.15-1283.84 mg/24 hours). After taking into account the subject's weight, the urinary excretion of Creatinine for the cuban subjects was lower than that of their anglosaxon counterparts. There was not correlation between the age of the subject and the urinary excretion of Creatinine. CONCLUSIONS: We recommend the use of the values of the urinary excretion of Creatinine tabulated in this article in the conduction of studies of body composition and nutritional assessment. PMID- 12723378 TI - [Interchangeability of the fat-to-fat-free mass ratios obtained by means arm anthropometric measures, skinfold thickness, and bioelectrical impedance]. AB - BACKGROUND: Body fat, fat free mass and the relationship between both, the fat-to fat-free mass ratio are useful parameters in the nutritional evaluation of the patient. The aim of this study is to evaluate the degree of agreement among the fat-to-fat-free mass ratios obtained by arm anthropometric measures (fat-muscle index, FMI), Siri equation for the sum of four skinfold thickness (body fat muscle index, BFMISiri) and triceps skinfold (BFMItriceps) and bioelectrical impedance (BFMIOmron) methods. METHODS: This is a cross-sectional study. A total of 145 patients were evaluated by anthropometry and bioelectrical impedance (Omron BF 300), being estimated the agreement through the Intraclass Correlation Coefficient (ICC) and Bland-Altman method. The reference method was BFMISiri. RESULTS: The ICC between BFMISiri-BFMItriceps were 0.9304 (0.9035; 0.9498), between BFMISiri-FMI of 0.7726 (0.6846; 0.8361) and between BFMISiri-BFMIOmron of 0.9114 (0.8771; 0.9361). BFMItriceps (limits of agreement -0.171 to 0.117) show the best agreement according to Bland-Altman analysis with BFMISiri, followed by BMFISiri-BFMIOmron (-0.186; 0.178). The agreement limits between FMI and BFMISiri (-0.2; 0.42), BFMItriceps (-0.26; 0.42) or BFMIOmron (-0.292; 0.504) were beyond of the established cut-off points (-0.2; 0.2). CONCLUSIONS: Due to the nature of the statistical agreement BFMItriceps and BFMIOmron are methods interchangeable methods between them and with BFMISiri. FMI is not interchangeable with BFMISiri, BFMIOmron or BFMItriceps. These results suggest that FMI cannot estimate properly the body composition. BFMItriceps and BFMIOmron are valid alternative methods to be used instead BFMISiri in the fat-to-fat-free mass ratio assessment. PMID- 12723379 TI - Daily intake of macronutrients in a group of institutionalized elderly people in Leon. Spain. AB - Monitoring of energy distribution of the three macronutrients of diet could be beneficial in order to improve the physiological status of elderly people. The objective of this study is to analyse total daily energy intake as well as the caloric contribution of the macronutrients and alcohol, which make up basic diet of five nursing homes in Leon (Spain). Dietary consumption was evaluated in a group of 107 elderly people, aged 65-98 years. A precise weighing method was used to conduct the control of food intake covering seven days. Protein, carbohydrates, fat, alcohol, dietary fiber and cholesterol intake were obtained. Weight, and Height also were measured. Total dietary energy intake was significantly higher in men (130.5%) than in women (115.6%), with regard to recommended value. Relative contribution of macronutrients to total energy intake is extremely unbalanced. Energy derived from protein was very high (16.7%), energy derived from fat was also very high, and significantly higher for females (39.6%) than for males (34.4%), whereas the proportion derived from carbohydrates was very low, although also significantly higher in females (41.5%) than in males (35.8%), due to the high energy percentage that make up the alcohol intake in males (9.1%). A review of the diet offered by nursing homes, not only directed at the adjustment of total energy intake but also with respect to alcohol intake and macronutrient content of foodstuffs used in the elaboration of the menus, would be required in order not to unbalance the caloric profile of the diet. PMID- 12723380 TI - [Nutritional status in head and neck cancer patients: the impact on the prognoses]. AB - Malnutrition is reported to affect 30-50% of all patients with head and neck cancer. The impact of malnutrition on surgical outcome is not clearly understood. The purpose of this study is to determine if nutritional status is related to postoperative complications (pharyngostoma) or tumor recurrence. METHODS: Fifty patients with T2-4 head and neck carcinomas underwent surgery were studied prospectively. Nutritional status was related to postoperative complications and 2-year survival. RESULTS: Twenty-seven patients present malnutrition (54%). We had 15 patients with pharyngostoma and 11 tumor recurrences. We did not find any correlation between the pharyngostoma and malnutrition, but we we found correlation between malnutrition and tumor recurrence (p < .05). CONCLUSION: Malnutrition has negative impact on outcome of patients with head and neck carcinomas. PMID- 12723381 TI - [Detection of malnutrition on admission to hospital]. AB - In view of the high prevalence of malnutrition in hospitals as shown by some studies and in the light of its direct impact on the increase in morbidity and mortality among hospitalized patients, it is necessary to ensure the early identification of this condition. This would allow appropriate nutritional treatment to be instituted as soon as possible for patients at risk who require it. The present multi-centric study included the participation of hospitals in the northern region of the country which have a Nutrition Unit, with use of the Overall Subjective Assessment and a few basic parameters. The study analyzed the nutritional status on admission of 620 patients hospitalized in the Internal Medicine and General Surgery areas during the months of December, 1999, January, February and March, 2000. We detected moderate malnutrition or suspected malnutrition in 20% of the patients studied and severe malnutrition in 18.2%, with a greater incidence in Internal Medicine. Malnutrition was mainly evidenced by weight loss of more than 5% and a reduction in subcutaneous fat and muscle mass. In addition, there are functional limitations on performing everyday activities. Changes in dietary intake were present in 40.8% of the patients. The associated gastro-intestinal symptoms included anorexia, which affected 37.7% of the group studied. Malnutrition is most frequently linked with infectious diseases, problems in circulation and with the digestive apparatus. We feel that the Overall Subjective Assessment, together with the measurement of height and weight, should be used systematically with patients on admission, as it is a simple and effective method for the identification of patients with nutritional risk. PMID- 12723383 TI - Editing the British Journal of Psychiatry. PMID- 12723382 TI - The privilege of editing a scientific journal. PMID- 12723384 TI - Editing a journal in an era of publishing change. PMID- 12723385 TI - Conflict of interest and the credibility of medical journals. PMID- 12723386 TI - Editing Social Psychiatry and Psychiatric Epidemiology. PMID- 12723387 TI - World Psychiatry: trying to reach psychiatrists worldwide. PMID- 12723389 TI - [Circumstances of the interview]. PMID- 12723388 TI - [Michael Shepherd and his influence on contemporary psychiatry]. PMID- 12723390 TI - [Michael Shepherd interviewed by David Healy (London, June 1995)]. PMID- 12723391 TI - [Efficacy of the "VADO" approach in psychiatric rehabilitation: a controlled study]. AB - AIMS: Assessment of the efficacy of the rehabilitation approach that is recommended by the manual VADO (AAOS in English: Abilities Assessment and Objectives Setting) in schizophrenic syndromes. METHODS: Controlled trial, partly with individual randomisation. Centres were invited to recruit 10 patients who in most centres were randomly allocated either to the VADO approach or to usual rehabilitation practice. At least two professionals for each centre attended a brief intensive training course. Patients were assessed at baseline and six months later with a) the FPS scale, which is an improvement on the DSM-IV SOFAS; b) the BPRS 4.0; c) AR, a clinical tool that is contained in VADO. RESULTS: 55 patients received the experimental interventions and 40 the control one. Six months later greater significant improvements were observed in the experimental group, both in social functioning and in psychopathology. CONCLUSIONS: The dissemination of structured rehabilitation approaches that are based on personalized definition of objective, intensive progress monitoring, patient's therapeutic education and validated strategies and do not imply additional costs, may be warranted. A longer follow-up is under way. PMID- 12723392 TI - [HoNOS-Rome: an extended, customized, and longitudinal oriented version of the HoNOS]. AB - AIMS: To clarify the acceptability, reliability and factorial validity of a new Italian version of the HoNOS called HoNOS-Rome. Its main innovations are both in design and in contents. METHODS: Face validity was assessed by surveying 3 focus groups. Reliability was assessed in 8 different pairs of raters on a sample of 24 patients; construct validity was analysed by factor analysis using a sample of 187 patients at 6 day centres. Acceptability was investigated by means an anonymous questionnaire filled by professionals that were using the instrument. RESULTS: Time of completion was low (range 4-12 minutes), the tool proved very acceptable and the reliability was good (weighted kappa > or = 0.71 for all items). Factor analysis was consistent with the division of HoNOS-Rome into four sensible factors accounting for 52% of the total variance. CONCLUSIONS: The findings indicate that HoNOS-Rome has a satisfactory degree of acceptability, construct validity and reliability, and may promote the routine evaluation of outcomes in mental health services. PMID- 12723394 TI - [The latest diagnosis and treatment on deep-seated mycosis. Expectations for micafungin, the first candin in Japan (discussion)]. PMID- 12723393 TI - [Validation of the Italian version of the Self-report Insight Scale]. AB - AIMS: To evaluate the psychometric properties of the Italian version of the Insight Scale (IS), self-report questionnaire assessing the awareness of psychiatric illness. The instrument contains two forms, the first A that enquires about the present status, and the second B that concerns past episodes of illness. METHOD: Factorial structure, internal consistency and concurrent validity (towards three selected items of the 24-item BPRS, Unusual thought content", Conceptual disorganization and Uncooperativeness) were studied on 80 chronic subjects affected by schizophrenia. Differences between acute and stabilised patients were investigated. Test-retest reliability was assessed in a sub-sample of 22 stable cases. RESULTS: The Italian IS showed satisfactory concurrent validity and reliability. Acute patients had lower scores than stabilised ones. Factorial analysis brought to the distinction between insight for need for care in the present and in the past, which seems both plausible and clinically-useful. CONCLUSION: The use of the IS Italian version may be encouraged as a valid insight self-report instrument. Sensitivity to change and predictive power concerning clinical and social outcome and adherence to treatment should be investigated. PMID- 12723395 TI - [A questionnaire survey on the theory of postoperative infection prophylaxis in otorhinolaryngology]. AB - A questionnaire survey on postoperative infection prophylaxis was conducted to achieve the consensus on the perioperative antimicrobial use among otolaryngologists in Japan during the period of time from April to July 2000. Fifty-two out of 84 otolaryngologists replied, and the following consensus was obtained. An antimicrobial prophylaxis (AMP) agent should be chosen based on their efficacy against the pathogens expected to be contaminants, such as Staphylococcus spp., Pseudomonas aeruginosa and Bacteroides fragilis group; Use an AMP agent that achieves a bactericidal concentrations in both the serum and operating site. Use an AMP agent that has little unfavourable side effects. Newer agents should be considered as therapeutics for postoperative infections. Therapeutic antimicrobial agents having no cross-resistance to the AMP agents should be used, if postoperative infection is suspected or developed. The most commonly used agent for clean operations is cefazolin (CEZ), followed by cefotiam (CTM) and piperacillin (PIPC), in this order. For clean-contaminated operations, the most commonly used agent is CEZ, followed by flomoxef (FMOX) and CTM. PMID- 12723396 TI - [Alteration of antibacterial activity of tosufloxacin and various antibacterial agents against Streptococcus pneumoniae]. AB - The minimum inhibitory concentrations (MICs) of tosufloxacin (TFLX), levofloxacin (LVFX), ciprofloxacin (CPFX), gatifloxacin (GFLX), sparfloxacin (SPFX), azithromycin (AZM), cefteram (CFTM), cefdinir (CFDN) and cefpodoxime (CPDX) against 337 clinical isolates of Streptococcus pneumoniae isolated from Japanese hospital from 1997 to 2002 were investigated by agar plate method. The incidence of penicillin-susceptible S. pneumoniae (PSSP), penicillin-intermediate resistant S. pneumoniae (PISP), and penicillin-resistant S. pneumoniae (PRSP) in each year was studied, and the MICs of antibacterial agents against these strains were determined. As the results, the total incidence of PSSP, PISP, and PRSP was 51.0%, 40.4% and 8.6%, respectively. The incidences of PSSP from 1997 to 2002 were 46.0-55.9%, and were almost definite in each year. In quinolone antibiotics, the differences of antibacterial activity among TFLX, SPFX, and GFLX against PSSP, PISP, and PRSP, were not observed, and these 3 quinolones had potent antibacterial activity. Although CPFX and LVFX showed antibacterial activity as well as other quinolones by 2001, the CPFX-resistant or LVFX-intermediate resistant strains of PSSP were seen with 56.5% and 91.3% in 2002, respectively. Thirty percents of each PSSP, PISP, and PRSP strains were AZM-resistant strains. Such tendency of increase was recognized in PSSP. Against cephem antibiotics, the incidence of intermediate resistant and resistant strains was higher for PISP and PRSP than for PSSP. No difference in the incidence of resistant strains was noted among CFTM, CFDN, and CPDX. PMID- 12723397 TI - [Relationship between arbekacin-susceptibility and aminoglycoside-resistant gene of methicillin-resistant Staphylococcus aureus (MRSA)]. AB - The susceptibility to arbekacin (ABK) of methicillin-resistant Staphylococcus aureus (MRSA) was investigated to find out how it related to aac(6')/aph(2") gene. In 49 isolates of MRSA for which MIC of ABK ranged from 0.125 to 64 micrograms/ml, the MICs of ABK for 38 strains carrying aac(6')/aph(2") gene were widely distributed from 0.25 to 64, whereas those for 11 strains without that gene were all < or = 0.5 microgram/ml. Residual rate of ABK activity was higher than that of gentamicin after the reaction with each crude enzyme preparation extracted from 3 isolates of MRSA, carrying aac(6')/aph(2") and aad(4',4") genes. Furthermore, 97 strains of MRSA isolated at Kanagawa prefecture in Japan in 1999 were all sensitive to ABK, although 28 strains of them carried aac(6')/aph(2") gene. These results showed that ABK resistance was not necessarily related to carrying aac(6')/aph(2") gene in clinical isolates of MRSA. PMID- 12723398 TI - [Attenuation of arbekacin-induced nephrotoxicity in rats by pazufloxacin mesilate]. AB - The protective effect of pazufloxacin (PZFX) mesilate, a parenteral quinolone antimicrobial agent, on arbekacin (ABK)-induced nephrotoxicity was evaluated with 8-week-old male Sprague-Dawley rats. Animals were injected with ABK at a dose of 32 mg/kg intramuscularly, or a combination of ABK in the same manner with PZFX mesilate at a dose of 208 mg/kg (160 mg/kg convert to PZFX, active principle of PZFX mesilate) intravenously once a day for 4 days. In consequent, ABK induced increases in protein, beta 2-microglobulin and N-acetyl-beta-(D)-glucosaminidase in urine, and histopathological phospholipidosis in kidneys. The extent of these changes was reduced when ABK was given in a combination with PZFX mesilate. Renal cortex level of ABK increased after an administration of ABK 1 hour to 4 hours; however, the increase was suppressed by coadministration of PZFX mesilate. Taken together, these results suggest that PZFX mesilate has the protective effect on ABK-induced nephrotoxicity, and that this was attributable to a suppression of uptake of ABK in cortical renal tubules. PMID- 12723399 TI - [Successful combination therapy of teicoplanin with cefmetazole on postoperative cardiovascular MRSA infections: clinical report on two cases]. AB - We clinically evaluated the efficacy of the combination therapy of teicoplanin (TEIC) with cefmetazole (CMZ) in two patients, one (case 1) had developed surgical site infection and the other (case 2) mediastinitis caused by methicillin-resistant Staphylococcus aureus (MRSA) after cardiovascular surgeries. TEIC (400 mg twice a day on day-1 and 400 mg once a day thereafter) was administered intravenously immediately after the end of drip infusion of CMZ (1.0 g twice a day). Both patients showed marked improvement on the 5th-day. The isolated MRSA from these two cases were subjected to the in vitro studies and synergistic effects between TEIC and CMZ were recognized under the checkerboard method. Thus, the favorable effects of the combination therapy might be attributable to the synergy between TEIC and CMZ recognized in vitro. These results suggest that the combination therapy of TEIC with CMZ may be useful in the treatment of severe MRSA infections. PMID- 12723401 TI - [Synthetic studies of microbial natural products from microorganisms]. PMID- 12723400 TI - [Effect of prophylaxis against myocosis in patients with hematological malignancy disease: efficacy of dosage of itraconazole]. AB - Prophylactic effect of itraconazole (ITCZ) in mycosis associated with hematological malignancy was studied. In a total of 12 patients of hematological malignancy admitted to Hokkaido University Hospital from April, 2001 to March, 2002, six patients received 100 mg per day of ITCZ and the other six received 200 mg per day of ITCZ. ITCZ was administrated from the starting of chemotherapy, and from the time point capable of oral administration in bone marrow transplantation patients. ITCZ administration was finished based on docter's judgement. As the results of this study, three cases with 100 mg per day of ITCZ had mycosis, but none with 200 mg per day of this drug had it. Mild itching associated with ITCZ was found only one case. In conclusion, we suggest that ITCZ is effective and safe drug in terms of prophylaxis of mycosis. PMID- 12723402 TI - [Suppressive effects of macrolides and their derivatives on quorum sensing systems in Pseudomonas aeruginosa]. PMID- 12723403 TI - [Analysis of nasopharyngeal flora in children with acute otitis media attending a day care center]. PMID- 12723404 TI - Clinical experience that many periodontists have had when treating poorly controlled diabetic patients. PMID- 12723405 TI - Immediate implant placement and provisionalization in edentulous, extraction, and sinus grafted sites. AB - The incorporation of restorative procedures during implant placement, as well as during the creation of natural emergence profiles and lifelike ceramic restorations, has become the focus of implantology over the last few years. Recent publications have provided guidelines for success with the immediate restoration procedure and have presented basic surgical protocols for the implant team. Enhancement of the healing phase through the local delivery of growth factors to the surgical site, as well as through advancements in bone grafting materials, has allowed the implant surgeon to accomplish multiple surgical procedures during the initial surgical visit. In addition, advancements in surgical stent designs have allowed the restorative dentist to adequately communicate to the surgeon during surgery the parameters required in the final restoration to replace the natural tooth system with form, function, and esthetics. This article presents the results of more than 400 immediate restored implants placed in edentulous sites, fresh extraction sockets, and sinus grafted sites. Also highlighted are guidelines for surgical success, as well as a description of a surgical stent design that communicates requirements for restorative success to the surgeon, while also serving as an esthetic provisional restoration. PMID- 12723406 TI - If not amalgam, then what? PMID- 12723407 TI - The efficacy of minocycline microspheres in the treatment of chronic periodontitis: statistical vs clinical significance. AB - This article addresses the benefits of using minocycline microspheres as an adjunct to conventional periodontal therapy. The author reviewed data from a large controlled clinical trial. The investigation indicated that scaling and root planing, plus minocycline microspheres, attained statistically significant improvements, with regard to several clinical parameters, when compared with scaling and root planing alone. Because it is the clinical meaningfulness of data that determines whether a therapy should be implemented, these data are interpreted with respect to their clinical relevance. PMID- 12723408 TI - How direct bonding satisfies an esthetic need. AB - The patient was more than satisfied with his appearance and with the cost of the procedure. After the wedding, he was ready to begin a more conventional series of treatments with orthodontia. Within the past several years, we have significantly progressed in our ability to provide complete in-office, long-term comprehensive dentistry. This article was written to help dentists visualize the potential of both materials and techniques for direct, in-mouth restorative dentistry. As more composites or composite replacement materials become available, practitioners will learn to perform direct-placement techniques even more skillfully. The majority of patients appear to covet white, bright teeth--sometimes to a degree that is beyond their normal color. This patient was no different. As a dentist, I have learned to give in to patients' demands. After all, it is their teeth. PMID- 12723409 TI - Osseous site development for optimal implant placement. PMID- 12723410 TI - Terrorism in the courtroom. PMID- 12723411 TI - "Quid pro quo" not the way to go. PMID- 12723412 TI - Bursting the bubble. PMID- 12723413 TI - Find the real enemy. PMID- 12723414 TI - Remember Osler's words. PMID- 12723415 TI - Parity doesn't equal higher costs. PMID- 12723416 TI - Professionalism redux. PMID- 12723417 TI - What's happening with HIPAA. PMID- 12723418 TI - Fat kids on the block. PMID- 12723419 TI - Mo' better blues. PMID- 12723421 TI - Medicine's changing face. PMID- 12723420 TI - Putting patients on trial. PMID- 12723422 TI - Born to be a doctor. Interview by Larry Besaw. PMID- 12723423 TI - Deaths of Texas adolescents from injury, 1996 through 1998. AB - From studying the deaths of Texas adolescents from injury during 1996 through 1998, we compiled data showing major causes. Injuries accounted for 3760 deaths and 75% of all adolescent deaths. Unintentional injuries (accidents) accounted for 65% of injury deaths and 49% of all deaths. Motor vehicle traffic accidents accounted for 74% of deaths from unintentional injuries and 36% of all deaths. Homicide accounted for 18% of deaths from injury and 14% of all deaths. Suicide accounted for 15% of deaths from injury and 11% of all deaths. The major single cause of adolescent death, assault with unspecified firearm, accounted for 483 deaths, 13% of injury deaths, and 10% of all deaths. Several age, gender, and ethnic differences were identified in risk of death from the various causes of injury, and these analyses may help target specific populations of Texas adolescents for strategies to prevent injuries. PMID- 12723424 TI - Cause-specific mortality among employees of a Texas-based chemical manufacturing facility, 1940 through 1996. AB - We identified and followed 27,795 Texas-based employees of The Dow Chemical Company from 1940 through 1996. Overall, the study population experienced favorable mortality patterns when compared with external populations. We observed lower death rates for several major causes of death, including heart disease and many cancers, which may be indicative of the "healthy worker effect" and the absence of major health hazards from these manufacturing facilities. Previous studies of the chemical company found higher-than-expected rates of lung, kidney, and brain cancer. More lung cancer deaths than expected continue to occur when the plant population is compared with the US and Texas populations but not with the local 5-county region. The numbers of brain and kidney cancers were also greater than expected, but the risk is attenuating. These findings, taken together with previously reported studies that examined these lung, kidney, and brain cancers relative to exposures to specific hazardous agents, do not suggest an occupational etiology. PMID- 12723425 TI - Larger than whose life? The 2003 MDA president's interview. PMID- 12723426 TI - Diagnosis and management of oral lichen planus. AB - Oral lichen planus is a complex and poorly understood clinical condition which cannot be cured. A definitive diagnosis and careful, conscientious follow-up are imperative. Symptoms and complications are common and challenging but may be managed with a variety of therapies including orally administered and systemic medications as well as lifestyle alterations and reduction of precipitating factors. PMID- 12723428 TI - Ambulatory BP monitoring. PMID- 12723429 TI - Will sex merchants invade your Web site? PMID- 12723427 TI - This side of the sea of tranquility. PMID- 12723430 TI - How do your staff salaries stack up? PMID- 12723431 TI - The target of a "shotgun" suit fires back. PMID- 12723432 TI - Add services without big debt. PMID- 12723433 TI - The key to a HIPAA-safe computer system. PMID- 12723434 TI - "My first inclination was to hit him in the mouth". PMID- 12723435 TI - Traveling safely in an unsafe world. PMID- 12723436 TI - Don't quit your day job. PMID- 12723437 TI - Credentialing made easy. PMID- 12723438 TI - Carpe diem, my new mantra. PMID- 12723439 TI - 16 Rx safeguards that also safeguard you. PMID- 12723440 TI - What's your practice worth? PMID- 12723441 TI - Moves you can make to cut premiums. PMID- 12723442 TI - Our adolescents--our future. PMID- 12723443 TI - Sex education in the schools: what role does it play? AB - Information regarding pubertal development, sex, pregnancy, and contraception should be provided to children and adolescents in an age-appropriate manner from parents, health care providers, and schools. This article reviews the medical literature on school sex education programs, adolescents' perception of these programs, and the role played by the media, parents, and health care professionals in sexual education of teens. PMID- 12723444 TI - Parenting adolescents. AB - The period of adolescence is often thought to be one of intense stress and turmoil. Yet many parents and teens negotiate this developmental stage without extreme family conflict and without sacrificing close relationships. This review summarizes a portion of the literature on parent-adolescent relationships, focusing on monitoring and control of adolescent behavior and parenting style. Basic principles to emphasize when working with adolescents and parents are also included. PMID- 12723445 TI - Clinical factors in group psychotherapy for parents of adolescents with disruptive behaviour disorders. AB - This paper describes a group treatment approach to helping parents of youth with Disruptive Behaviour Disorders. These youth have a significant negative and pervasive impact on their parents, who feel incompetent and guilty. Parents are helped to express their grief at their many losses in a safe, supportive and non blaming group environment. They gain insight into the factors that make it difficult for them to set limits and they begin to change themselves rather than "fix" their child. Parent's hope and confidence grow as positive change becomes evident. As their sense of shame, guilt and fear lessen, parents are able to act more effectively with their youth and others. This paper delineates the clinical factors that contribute to parental effectiveness. PMID- 12723446 TI - Evaluation of the parent group experience. What helps and what hinders. AB - This paper describes the responses of parents to questionnaires filled out as part of a group therapy program for parents whose adolescents exhibit conduct problems and refuse interventions. A brief description of the youth, parents and the program is followed by representative examples of the comments parents made at their termination from the group about their experiences and what helped them or hindered them in dealing with their situations and emotions. Their responses are compared and contrasted with the responses to Irving Yalom's adult groups responses to a Q-sort, which rated aspects of their groups from most helpful to least helpful. Parents' experiences are an informative guide as to their perceptions and what they feel helped or hindered them in this program. PMID- 12723447 TI - A longitudinal study of parenting and adolescent adjustment in Chinese adolescents with economic disadvantage. AB - This longitudinal study examines the relationship between parenting behavior and adolescent adjustment (psychological well-being, substance abuse and delinquent behavior) in Chinese adolescents with economic disadvantage (N = 199). Results showed that parenting characteristics were concurrently and longitudinally related to measures of adolescent adjustment, particularly adolescent problem behavior. Compared with the norm based on adolescents of a community sample, poor adolescents perceived parenting characteristics to be more negative and they had relatively lower life satisfaction. Paternal parenting was perceived to be more negative than maternal parenting and parenting behavior was perceived to deteriorate over time. PMID- 12723448 TI - Parenting style of Chinese fathers in Hong Kong: correlates with children's school-related performance. AB - This study investigates parenting styles among Chinese fathers in Hong Kong as perceived by their school-age children. Four parenting styles, namely inductive, indulgent, indifferent, and dictatorial parenting, are assessed using the Parent Behavior Report (1988). Data were collected through a questionnaire survey on a sample of 1011 Primary Three to Five Chinese students from six schools in Hong Kong and 471 fathers. Findings show that among Chinese fathers, the least common parenting style is inductive, while the other three styles are of similar occurrence. Chi-square analysis shows no significant association between children's grade level and father's parenting style. However, there is a significant association with gender, with fathers more likely to be perceived as dictatorial with boys and indulgent with girls. The effect of paternal styles on children's school-related performance is also examined. MANOVA results show that significant differences are found among children of the four paternal style groups with respect to academic performance, interest in school work, aspiration for education, involvement in extracurricular activities, and efficacy for self regulated learning. Post-hoc tests reveal that children's performance is similar between the groups with indulgent and inductive fathers, and between children of indifferent and dictatorial fathers, with the former groups performing better than the latter in general. Findings are discussed with regard to research on parenting style and paternal behavior, as well as understanding the roles of fathers in Chinese families in the socio-cultural context in Hong Kong. PMID- 12723449 TI - Covert parental control: parent-adolescent interaction and adolescent development in a Chinese context. AB - Although there are well-known theories of adolescent development in the West, there is a notable lack of theory and empirical support for understanding the process of Chinese adolescent development. This paper investigates the parental attitudes and practices of Chinese-Canadian immigrant parents and the reactions of adolescents in an effort to elucidate the pattern of parent-adolescent interaction and the process of adolescent development in the Chinese-Canadian cultural context. A qualitative study approach was used to explore the experiences and views of 19 adolescents and ten of their parents. The findings indicate that Chinese parents and adolescents co-construct the dominant theme of "covert parental control" in the adolescent development process. "Covert control," in this context, refers to a form of parenting characterized by parental guidance, family teaching, coaching, and monitoring, Parents, through "tact" and "skills," successfully influence and guide their adolescent children. Adolescents, motivated by their sense of loyalty to family and the concepts associated with parental guidance, reinforce their parents' covert control and the indigenous concept of "guan" in the practice of Chinese parenting. PMID- 12723450 TI - Educating adolescents about puberty: what are we missing? AB - Adolescents undergo significant physical and cognitive changes during their pubertal development. These changes contribute to and impact their future development. Educating adolescents at an early age about their expected development decreases the possible anxiety associated with this period of life and also helps adolescents make better choices in regards to their sexuality. In order to assess the degree of education regarding pubertal development and sexuality, we conducted a survey of late adolescents (Median age 19 years) and parents of adolescents. A total of 409 adolescents (237 females, 172 males) and 124 parents completed the survey. 14.4% of teens (36.6% of males and 2% of females) reported that no one spoke to them prior or during puberty about pubertal development or sexuality issues. Teens receiving some form of puberty/sexuality education did so at a median age of 13 for girls and 15 for boys. More than one source of information was the most common (49%) followed by mother only (20%). 85% of parents reported talking to their teens about pubertal development and sexuality. There were several differences between areas reported covered by parents but not by teens, for example 72% of parents reported talking to their teens about gender differences in growth but only 31% of teens reported being spoken to about that. Areas that are very poorly covered are breast development in boys and sexual assault/date rape in girls at 5% and 26% respectively. In summary, it appears that we continue to do a relatively poor job in educating our kids about their development and sexuality and we do it late. Boys are even less likely than girls to be talked to about many areas of pubertal development and sexuality and when that is done, it is done at a later age. PMID- 12723451 TI - Barriers to home care and social support for an adolescent with disability. AB - Home care for children with disability has expanded in the last twenty years in order to support the child and family at home and avoid hospitalization or long term care facilities. A case story is presented to show several barriers to home care for a 15 year old boy with Hunter's syndrome and severe disability. The case showed a need for further education and coordination of professionals in order for pediatric home care to be more efficient and supportive in Israel. PMID- 12723452 TI - The life mission theory: a theory for a consciousness-based medicine. AB - Genetic factors, external stress and the human factor are influential to the health and well-being of every person. Several studies have shown that the human being have many internal powers that can promote health and increase quality of life. A theory on the human meaning of life is put forward and how it relates to health, disease and quality of life in the context of holistic medicine. PMID- 12723453 TI - Trends in adolescent sexual risk behaviors. AB - Many societies worldwide are faced with an increase in adolescent pregnancy, rape, sexual abuse and sexually transmitted disease due to different sexual behaviors and sometimes lack of responsible sexual behaviors. The World Health Organization and the Surgeon General of the United States Government have called for action to this serious public health issue. In this short communication data from the United States, Canada and Israel is presented with a call for a professional response to this public health challenge. PMID- 12723454 TI - Biomarkers in drug discovery and development: from target identification through drug marketing. AB - Biomarkers of disease play an important role in medicine and have begun to assume a greater role in drug discovery and development. The challenge for biomarkers is to allow earlier, more robust drug safety and efficacy measurements. Their role in drug development will continue to grow for the foreseeable future. For biomarkers to assume their rightful role, greater understanding of the mechanism of disease progression and therapeutic intervention is needed. In addition, greater understanding of the requirements for biomarker selection and validation, biomarker assay method validation and application, and clinical endpoint validation and application is needed. Biomarkers need to be taken into account while the therapeutic target is still being identified and the concept is being formulated. Biomarkers need to be incorporated into a continuous cycle that takes what is learned from the discovery and development of one series of biomarkers and translates it into the next series of biomarkers. Optimum biomarker development and application will require a team approach because of the multifaceted nature of biomarker selection, validation, and application, using such techniques as pharmacoepidemiology, pharmacogenetics, pharmacogenomics, and functional proteomics; bioanalytical method development and validation; disease process and therapeutic intervention assessments; and pharmacokinetic/pharmacodynamic modeling and simulation to improve and refine drug development. The potential for biomarkers in medicine and drug development will be limited by the least effective component of the processes. The team approach will minimize the potential for the least effective component to be fatal to the rest of the process. As scientific/regulatory foundations for biomarkers in medicine and drug development begin to be established, successes and applications will need to be effectively communicated with all of the stakeholders, including not only internal and external drug developers and regulators but also the medical community, to ensure that biomarkers are totally integrated into drug discovery and development as well as the practice of medicine. PMID- 12723455 TI - Pharmacogenetics and pharmacogenomics in drug development and regulatory decision making: report of the first FDA-PWG-PhRMA-DruSafe Workshop. AB - The use of pharmacogenetics and pharmacogenomics in the drug development process, and in the assessment of such data submitted to regulatory agencies by industry, has generated significant enthusiasm as well as important reservations within the scientific and medical communities. This situation has arisen because of the increasing number of exploratory and confirmatory investigations into variations in RNA expression patterns and DNA sequences being conducted in the preclinical and clinical phases of drug development, and the uncertainty surrounding the acceptance of these data by regulatory agencies. This report summarizes the outcome of a workshop cosponsored by the Food and Drug Administration (FDA), the Pharmacogenetics Working Group (PWG), the Pharmaceutical Research and Manufacturers of America (PhRMA), and the PhRMA Preclinical Safety Committee (DruSafe). The specific aim of the workshop was to identify key issues associated with the application of pharmacogenetics and pharmacogenomics, including the feasibility of a regulatory "safe harbor" for exploratory genome-based data, and to provide a forum for industry-regulatory agency dialogue on these important issues. PMID- 12723456 TI - Tegaserod pharmacokinetics are similar in patients with severe renal insufficiency and in healthy subjects. AB - Tegaserod (HTF 919), a selective 5-HT4 receptor partial agonist with promotile activity throughout the gastrointestinal tract, is in development for the treatment of irritable bowel syndrome. In an open-label, parallel-group study, the pharmacokinetics of a single 12-mg oral dose of tegaserod in patients with severe renal insufficiency requiring hemodialysis were compared with data obtained from healthy subjects matched for age, weight, height, and gender (n = 10, both). The pharmacokinetics of tegaserod were similar in both groups (AUC(0h tz), ng.h/ml: 14.6 +/- 8.5 vs. 14.3 +/- 7.1; Cmax, ng/ml: 4.6 +/- 2.3 vs. 5.1 +/- 2.2; tmax, h: 1.0, for both). Tegaserod had similar tolerability in renally impaired patients and healthy volunteers, with adverse events largely related to the gastrointestinal pharmacological actions of the drug. Therefore, no dose adjustment of tegaserod is necessary for patients with renal insufficiency. PMID- 12723457 TI - Population pharmacokinetics and pharmacodynamics of ciclesonide. AB - Ciclesonide is a novel glucocorticoid that is converted into ciclesonide--active principle (CIC-AP) in the lung. The study objectives were to identify a structural model for population pharmacokinetic (PK) analysis of CIC-AP using nonlinear mixed-effects modeling, assess the influence of select covariates on PK and/or pharmacodynamic (PD) parameters, and investigate the effects of CIC-AP on endogenous cortisol. Pooled concentration data from nine phase I studies (dose: 400-3600 micrograms) involving healthy and asthmatic patients were included in the PK analysis. There were 151 subjects (3300 observations) for the CIC-AP population PK analysis. Various models examined inter- and intrasubject variability for the PK parameters. Population estimates of the PK parameters of clearance and volume of distribution were 396 L/h (64.8% co-efficient of variation [CV]) and 1190 L (41.2% CV), respectively. Pharmacodynamic population estimates included maximum cortisol release rate, 3140 ng/h (5.4% CV). The EC50 of CIC-AP was 0.88 ng/mL. Ciclesonide is a safe corticosteroid that causes negligible cortisol suppression. The disposition and effect of CIC-AP can be described using mixed-effect modeling. The estimated EC50 is similar to mean Cmax from an 800-micrograms dose, further suggesting CIC-AP has little effect on cortisol suppression. PMID- 12723458 TI - Pharmacokinetics of an immediate-release oral formulation of Fampridine (4 aminopyridine) in normal subjects and patients with spinal cord injury. AB - Plasma concentration profiles of the K+ channel-blocking compound Fampridine were obtained from (1) control subjects (n = 6) following oral administration of doses of 10, 15, 20, and 25 mg and (2) patients with spinal cord injury (SCI) (n = 11) following a single oral dose of 10 mg of an immediate-release formulation. Plasma concentrations were determined using a reversed-phase ion-pair high-performance liquid chromatography (HPLC) assay with ultraviolet light detection employing liquid extraction. The drug was rapidly absorbed with a tmax approximately 1 hour for both groups; tmax was independent of dose. Cmax and AUC0-infinity were linearly related to dose, and t 1/2 was 3 to 4 hours for both groups. There were no obvious differences in the (10-mg) plasma concentration profiles between control subjects and SCI patients. The drug was well tolerated, with only mild and transient side effects of light-headedness, dysesthesias, and dizziness. PMID- 12723459 TI - Population kinetics, efficacy, and safety of dichloroacetate for lactic acidosis due to severe malaria in children. AB - The authors conducted a randomized, double-blind, placebo-controlled trial of intravenous dichloroacetate (DCA) for the purpose of treating lactic acidosis in 124 West African children with severe Plasmodium falciparum malaria. Lactic acidosis independently predicts mortality in severe malaria, and DCA stimulates the oxidative removal of lactate in vivo. A single infusion of 50 mg/kg DCA was well tolerated. When administered at the same time as a dose of intravenous quinine, DCA significantly increased the initial rate and magnitude of fall in blood lactate levels and did not interfere with the plasma kinetics of quinine. The authors developed a novel population pharmacokinetic-pharmacodynamic indirect response model for DCA that incorporated characteristics associated with disease reversal. The model describes the complex relationships among antimalarial treatment procedures, plasma DCA concentrations, and the drug's lactate-lowering effect. DCA significantly reduces the concentration of blood lactate, an independent predictor of mortality in malaria. Its prospective evaluation in affecting mortality in this disorder appears warranted. PMID- 12723460 TI - Pharmacokinetics and pharmacodynamics of a single bolus of propofol 2% in healthy volunteers. AB - This study was undertaken to assess the bioequivalence between a new formulation of propofol 2% and the commercially available product Diprivan. Secondary objectives were to compare the times to onset of and emergence from hypnosis, the hemodynamic effects, and the safety profiles. Twelve healthy male volunteers were included in a randomized crossover study. Subjects were administered a 2-mg/kg single bolus injection of each formulation separated by a 7- to 10-day washout period. Plasma propofol was determined by reversed-phase liquid chromatography with fluorescence detection. Eleven subjects completed the study, and both formulations were considered bioequivalent. There were no serious or severe adverse events. The concentration-time profiles of all the subjects could adequately be described using a three-compartment model. The mean times to cessation of counting out loud (17 vs. 18 s) and to eye opening (245 vs. 244 s) were not statistically different between treatment groups. Moreover, they seem to show some degree of pharmacodynamic bioequivalence, although a higher number of subjects are necessary to unequivocally demonstrate it. PMID- 12723461 TI - Single-dose pharmacokinetics and pharmacodynamics of RWJ 67657, a specific p38 mitogen-activated protein kinase inhibitor: a first-in-human study. AB - The objective of this study was to investigate the pharmacokinetics and ex vivo pharmacodynamics of increasing doses of RWJ 67657, along with the effect of food at one dose level in a first-in-human (FIH) study. This was a placebo-controlled, double-blind, randomized trial in healthy male subjects. Subjects received increasing doses of RWJ 67657 or placebo as a single oral dose (0.25-30 mg/kg) under fasting conditions. The effect of food was investigated for the 10-mg/kg dose. Plasma concentrations of RWJ 67657 were measured over a period of 48 hours using a validated LC-MS/MS method. To evaluate the pharmacodynamics of RWJ 67657, inhibition of cytokine production was monitored from exvivo-stimulated polymorphonuclear blood cells (PBMCs). Pharmacokinetic/pharmacodynamic modeling was used to characterize the inhibitory activity of RWJ 67657. RWJ 67657 was rapidly absorbed (mean tmax = 0.6-2.5 h). The pharmacokinetics of RWJ 67657 appear to be nonlinear with respect to single-dose administration of the investigative formulation. Coadministration of food did not have a significant effect on half-life or time to peak concentration (tmax) but decreased the exposure. Mean Cmax values in the presence of food were almost 50% lower than during fasting (542 vs. 1283 ng/mL), and the AUC decreased from 2832 to 1904 ng.h/mL with food. RWJ 67657 inhibited TNF-alpha, IL-8, and IL-6 in a concentration-dependent manner with mean IC50 values of 0.18 microM, 0.04 microM, and 0.43 microM, respectively. At 20 mg/kg, the median inhibition was greater than 85%. There were no significant adverse effects associated with single doses of this drug. This study demonstrates that RWJ 67657 has acceptable safety and pharmacokinetics to warrant further investigation in a repeat-dose setting. In addition, the early determination of effect on biomarkers suggests potential efficacy in diseases mediated by proinflammatory and inflammatory cytokines. PMID- 12723462 TI - Short-term exposure to low-dose ritonavir impairs clearance and enhances adverse effects of trazodone. AB - Antiretroviral agents may participate in drug interactions that influence the efficacy and toxicity of other antiretrovirals, as well as pharmacologic treatments of coincident or complicating diseases. The viral protease inhibitor, ritonavir, may cause drug interactions by inhibiting the activity of cytochrome P450-3A (CYP3A) isoforms. In a single-dose, blinded, four-way crossover study, 10 healthy volunteer subjects received 50 mg of trazodone hydrochloride or matching placebo concurrent with low-dose ritonavir (four doses of 200 mg each) or with placebo. Compared to the control condition, ritonavir significantly reduced apparent oral clearance of trazodone (155 +/- 23 vs. 75 +/- 12 ml/min, p < 0.001), prolonged elimination half-life (6.7 +/- 0.7 vs. 14.9 +/- 3.9 h, p < 0.05), and increased peak plasma concentrations (842 +/- 64 vs. 1125 +/- 111 ng/ml, p < 0.05) (mean +/- SE). Coadministration of trazodone with ritonavir increased sedation, fatigue, and performance impairment compared to trazodone plus placebo; differences reached significance only for the digitsymbol substitution test. Three subjects experienced nausea, dizziness, or hypotension when trazodone was given with ritonavir; 1 of these subjects also experienced syncope. Thus short-term low-dose administration of ritonavir impairs oral clearance of trazodone and increases the occurrence of adverse reactions. The findings are consistent with impairment of CYP3A-mediated trazodone metabolism by ritonavir. PMID- 12723463 TI - Interaction between ketoconazole and almotriptan in healthy volunteers. AB - The interaction between almotriptan, a 5-HT1B/1D agonist, and the potent CYP3A4 inhibitor ketoconazole was examined in 16 healthy volunteers. Subjects received (A) 12.5 mg almotriptan orally on Day 2 of a 3-day regimen of 400 mg ketoconazole once daily and (B) 12.5 mg almotriptan in a crossover design. Plasma and urine concentrations of almotriptan were measured by HPLC. Treatment effects on almotriptan pharmacokinetics were assessed by analysis of variance. Ketoconazole coadministration increased mean almotriptan AUC and Cmax from 312 to 490 ng h/mL and 52.6 to 84.5 ng/mL, respectively. Mean oral clearance was decreased from 40.7 to 26.2 L/h by ketoconazole, with an accompanying increase in the fraction of almotriptan excreted unchanged in the urine (40.6% to 53.3%) and a decrease in renal clearance (16.4 to 13.8 L/h). These effects were statistically significant. The effects of ketoconazole on almotriptan clearance were consistent with inhibition of the CYP3A4-mediated metabolism and a slight effect on the active tubular secretion of almotriptan. PMID- 12723464 TI - Effects of orlistat, a lipase inhibitor, on the pharmacokinetics of three highly lipophilic drugs (amiodarone, fluoxetine, and simvastatin) in healthy volunteers. AB - To investigate the effect of orlistat on the pharmacokinetics of three highly lipophilic drugs (amiodarone, fluoxetine, and simvastatin), the authors performed double-blind, placebo-controlled, randomized two-period crossover (for fluoxetine and simvastatin) or parallel (for amiodarone) studies in healthy volunteers ages 18 to 65 years of a body mass index between 18 and 30 kg/m2. During treatment with orlistat or matching placebo for 5 to 13 1/3 days, a single oral dose of highly lipophilic drug was administered, followed by obtaining serial blood samples for measuring plasma (for fluoxetine and simvastatin) or serum (for amiodarone) concentrations of the lipophilic drug and its active metabolite. Treatments were compared for the pharmacokinetic parameters AUC0-infinity, Cmax, tmax, and t 1/2 of highly lipophilic drugs and active metabolites. Analysis of variance was performed to assess the significance of the sequence effect and provide the variance estimate for the 90% confidence intervals. Subjects were also evaluated for adverse events, vital signs, and clinical and laboratory safety. The absorption of amiodarone (and active metabolite) was significantly reduced by approximately one-quarter using parameters of Cmax and AUC, while no inhibition of absorption was observed for fluoxetine and simvastatin as well as their active metabolites. There were no clinically significant differences in t 1/2 and tmax for all three drugs tested. Due to expected gastrointestinal adverse events known to occur with orlistat, there was a higher incidence of adverse events under regimen B (highly lipophilic drugs and orlistat) than under regimen A (highly lipophilic drugs and placebo). Other adverse events were sporadic and unremarkable. There were no clinically relevant changes in vital signs or laboratory values. In conclusion, except for amiodarone, there was no effect of orlistat on the pharmacokinetics of highly lipophilic drugs when these drugs were taken concomitantly with orlistat. PMID- 12723465 TI - [The anniversary of DNA: double-stranded DNA is 50 years old]. PMID- 12723466 TI - [Genetic defects as markers of tumor development]. AB - Carcinogenesis is long-term multistep accumulation of defects of genes responsible for cell division, DNA repair, and apoptosis. The functions of these genes are known both for norm and for pathologies caused by their damage and resulting in "asocial" cell behavior. Owing to the recent progress in studying the mechanisms of carcinogenesis, some genetic defects may be considered from the applied point of view (as tumor markers rather than as pathogenetic factors) and employed in diagnostics. Thus detection of mutant alleles in biological fluids (e.g., beyond the tumor) suggests higher risk of carcinogenesis. Genetic defects are a new class of tumor markers and have a substantial diagnostic potential. In contrast to known protein markers (alpha-fetoprotein, etc.) used in clinical practice, DNA markers are oncospecific (as these are in direct cause-and-effect relationships with carcinogenesis) and universal (as there is not a single tumor cell without a genetic defect). Analysis of DNA markers may be employed not only in diagnostics or tumor growth monitoring (assessment of treatment efficiency, early detection of recurrence or metastasis), but also (prospectively) in screening (tumor detection at the presymptomatic stage, identification of high risk groups). Theoretical grounds, prospects, problems, and methods of this new field are considered. PMID- 12723468 TI - [Segment duplications in the human genome]. AB - The review considers the structure, evolution, and possible mechanisms of spreading of intrachromosomal and interchromosomal segment duplications (SD), which account for more than 5% of the human genome. Most SD are mosaic and consist of multiple modules, which occur in several copies in different genome regions. SD are preferentially located in pericentric and subtelomeric regions, which are least studied on the human chromosomes. Homologous recombination between SD results in various chromosome rearrangements, contributing to the genome instability and the origin of several human hereditary disorders. PMID- 12723467 TI - [New tumor suppressor genes in hot spots of human chromosome 3: new methods of identification]. AB - Studies of the recent decade, including sequencing of numerous human genome regions, allowed a great progress in detection of new tumor suppressor genes (TSG) and development of new means of their identification and analysis. Effective methods of genome scanning and TSG identification combine DNA array techniques and subtraction hybridization. Alternative ways take advantage of new extrachromosomal vector systems (pETE, pETR) and the functional gene inactivation test. A breakthrough was made in localizing new TSG on the human chromosome 3 short arm, which harbors tumor-suppressing regions and is often rearranged in various tumors and in early carcinogenesis. On 3p, only three putative TSG were known five years ago, and at least ten were identified by the end of 2002. The role of new TSG in carcinogenesis is commonly inferred from a decrease in their transcription in tumor cell lines or primary tumors and from their ability to suppress the growth of these. Protein products of 3p TGS play an important part, constraining cell malignization. Some are directly involved in regulating the cell cycle and inducing apoptosis (RASSFIA), others suppress angiogenesis (Sema3B) or metastasis (Hyal-1). Numerous attempts to find mutations in exons of silent genes failed, and at least half of the new candidate genes (RASSFIA, CACNA2D2, BLU, HYAL1, SEMA3B, RAR-beta) proved to be inactivated by promoter methylation. PMID- 12723469 TI - [Segment duplications in subtelomeric regions of human chromosome 13]. AB - Owing to a great progress in studying the human genome, its euchromatic portion is almost completely sequenced; the complete sequence is still unknown only for pericentric and telomeric regions and short arms of acrocentric chromosomes. Extended satellite blocks and segment duplications located in these regions substantially hinder the joining of the sequenced fragments and construction of the full-length genome map. The sequence was established for a 1.5-kb human chromosome 13 subtelomeric region, which is about 10 kb away from the rDNA cluster, and deposited in GenBank under accession no. AF478540. The region showed 83-84% homology to the pericentric region of human chromosome 19, and contained short fragments homologous to the pericentric region of human chromosome 13. The results may contribute to the current revision of genome evolution concepts in view of numerous segment duplications revealed. PMID- 12723470 TI - [Tandem and interspersed repeats contribute to the mosaic structure of segmented duplications in the human genome]. AB - Intrachromosomal and interchromosomal segmental duplications account for more than 5% of the human genome. To analyze the processes resulting in the complex mosaic structure of duplicons, a draft human genome sequence was searched for duplicated segments of a genomic fragment of the pericentric region of the chromosome 21 short arm. The duplicons found consist of modules having paralogs in various genome regions. Module ends are flanked with various tandem or interspersed repeats, which are more unstable as compared with unique sequences. In most cases, the boundaries of duplicated segments exactly coincide with or are in close proximity to hot spots of various rearrangements within repeats or boundaries between repeats and unique sequences or between two different repeats. Homologous recombination between repetitive elements was assumed to be the major mechanism contributing to the mosaic structure of duplicons. PMID- 12723471 TI - [Genetic peculiarity of the yakut population from the autosomal loci data]. AB - The autosomal gene pool of Yakuts was analyzed with a panel of polymorphic Alu insertions. The observed allele frequencies were typical for other Asian ethnic groups. Genetic differentiation of three Yakut populations was relatively high, 2%. East Siberian ethnic groups were shown to have a common gene pool and to experience no intense gene flow from other populations. Development of the Yakut gene pool was assumed to involve no substantial genetic effect of neighboring populations. The results fit both autochthonous and southern origin hypotheses. PMID- 12723472 TI - [Methods of computer modeling and conformational mobility of DNA duplexes]. AB - The review is focused on issues of transferability of the context-sensitive conformational characteristics of DNA estimated from crystallographic structural data on the DNA in aqueous solution. The state of the art in molecular dynamics of charged biopolymers in aqueous solution is covered. Elaboration of expedient force fields and algorithms of calculating long-range electrostatic interactions and solving combined equations of atomic motion have made it possible to generate stable nanosecond trajectories of thermal atomic motion of the biopolymer in aqueous solution in the presence of counterions and salt ions over reasonable time. Tools for analyzing the atomic statistical trajectories of DNA duplexes in aqueous solution to infer context-sensitive conformational dynamic characteristics are discussed together with advances in simulating the mechanisms of global axial bend in DNA duplexes. These techniques allow one to consecutively analyze relationships between the contextual composition of the duplex and the basic modes of essential motions, their amplitude and extent of fluctuation. Development of satisfactory methods for estimating the free energy of biopolymer conformations in solution permits qualitative assessment of the conformational thermodynamic stability of biopolymers and their complexes. PMID- 12723473 TI - [Parallel-stranded DNA with natural base sequences]. AB - Noncanonical parallel-stranded DNA double helices (ps-DNA) comprising natural nucleotide sequences are usually second in stability to antiparallel-stranded (aps) canonical DNA structures, which ensures reliable cell functioning. However, recent data indicate a possible role of ps-DNA in DNA loops or in trinucleotide repeats connected with neurodegenerative diseases. The review surveys recent studies on the effect of nucleotide sequence on preference of one or other type of DNA duplex. (1) Ps-DNA with mixed AT/GC composition was found to have conformational and thermodynamic properties drastically different from those of Watson-Crick double helix. Its stability depends strongly on the specific sequence in a manner peculiar to the ps double helix, because of the energy disadvantage of the AT/GC contacts. The AT/GC boundary facilitated flipping of A and T out of the ps double helix. Proton acceptor groups of bases are exposed into the both grooves of the ps-DNA and are accessible to solvent and ligands, including proteins. (2) DNA regions containing natural minor bases isoguanine and isomethylcytosine were shown to form ps-DNA with transAT-, trans isoGC, and trans iso5meCG pairs exceeding in stability a related aps duplex. (3) Nucleotide sequence dG(GT)4G from yeast telomeres and microsatellites was demonstrated to form novel ps-DNA with GG and TT base pairing. Unlike d(GT)n and d(GnTm) sequences able to form quadruplexes, the dG(GT)4G sequence formed no alternative double- or multistranded structures in a wide range of experimental conditions, thus suggesting that the nucleotide context governs the observed structural polymorphism of the d(GT)n sequence. The possible biological role of ps-DNA and the prospects of its study are discussed. PMID- 12723475 TI - [Conformational effects of DNA stretching]. AB - DNA is an extensible molecule, and an extended conformation of DNA is involved in some biological processes. We have examined the effect of elongation stress on the conformational properties of DNA base pairs by conformational analysis. The calculations show that stretching does significantly affect the conformational properties and flexibilities of base pairs. In particular, we have found that the propeller twist in base pairs reverses its sign upon stretching. The energy profile analysis indicates that electrostatic interactions make a major contribution to the stabilization of the positive-propeller-twist configuration in stretched DNA. This stretching also results in a monotonic decrease in the helical twist angle, tending to unwind the double helix. Fluctuations in most variables initially increase upon stretching, because of unstacking of base pairs, but then the fluctuations decrease as DNA is stretched further, owing to the formation of specific interactions between base pairs induced by the positive propeller twist. Thus, the stretching of DNA has particularly significant effects upon DNA flexibility. These changes in both the conformation and flexibility of base pairs probably have a role in functional interactions with proteins. PMID- 12723474 TI - [Definitive role of polar residue clusters in B-DNA major groove recognition by protein factors]. AB - The 3D structural data for a number of protein-DNA complexes were used to analyze the regions of specific contact with the major groove of B-DNA double helix. The set included seven nonhomologous complexes featuring 12 DNA-binding domains of transcription factors and regulatory factors. The protein domains differed in structure, contained different motifs in the binding region, and broadly varied in chain length, from 30 to 200 residues. Protein-DNA interaction was assessed as hydrogen bonding between polar atoms and van der Waals contacts between nonpolar atoms. The binding sites were formed mainly through polar side chain interactions. On average, the recognition site comprises seven residues, six of them polar. The contact residues nearly always belong to a largest polar cluster of the protein. Thus one can think that the protein polar residue clusters play an important role in forming the protein-DNA recognition module. PMID- 12723476 TI - [Replication of the baculovirus genome]. AB - The review describes the current state of studying the baculovirus DNA replication. The structural organization of replication initiation sites and replication intermediates are considered. Attention is focused on virus replication factors, including DNA polymerase, helicase, IE-1, LEF-1, LEF-2, and LEF-3. PMID- 12723477 TI - [Prokaryotic DNA methyltransferases: the structure and the mechanism of interaction with DNA]. AB - The review considers current views on the function of DNA methyltransferases (MTases) that belong to prokaryotic type II restriction-modification systems. A commonly accepted classification of MTases is described along with their primary and tertiary structures and molecular mechanisms of their specific interaction with DNA (including methylation). MTase inhibitors are also considered. Special emphasis is placed on the flipping of the target heterocyclic base out of the double helix and on the methods employed in its analysis. Base flipping is a fundamentally new type of DNA conformational changes and is also of importance in the case of other DNA-operating enzymes. MTases show unique sequence homology, and are similar in structure of functional centers and in the mechanism of methylation. These data contribute to the understanding of the general biological significance of methylation, since prokaryotic and eukaryotic MTases are structurally and functionally similar. PMID- 12723478 TI - [Structural and functional analyses of the Hfb1, Hmob3 and Hmob33 cDNAs as an example of human brain-specific gene studies]. AB - Brain-specific human genes were studied over the recent years in the Department of Molecular Bases of Human Genetics, Institute of Molecular Genetics. Clones Hfb1, Hmob3, and Hmob33 were selected from human brain cDNA libraries by differential screening. The clones were sequenced, mapped, and tested for expression in various human tissues. In vitro and in silico experiments identified Hfb1 as an earlier unknown complexin 2 gene (CPLX2) fragment, which codes for the large 3'-untranslated region of the CPLX2 mRNA. Hmob3 proved to correspond to an earlier unknown fragment of the large 3'-untranslated region of the human MAP1B mRNA. With Hfb1 and Hmob3, new terminal exons were revealed and exact structures established for CPLX2 and MAP1B. Hmob33 was identified as a fragment of the 3'-terminal exon of a new gene, MOB, which codes for a thus far unknown evolutionarily conserved transmembrane protein. The structure of the deduced protein product was analyzed. PMID- 12723480 TI - [Molecular construction (superstructures) with adjustable properties based on double-stranded nucleic acids]. AB - Formation of molecular construction that consists of double-stranded molecules of nucleic acids (or synthetic polynucleotides) located at the distance 30-50 A in the spatial structure of particles of their cholesteric liquidcrystalline dispersion and crosslinked by polymeric chelate bridges is described. The resulting superstructure, which possesses peculiar physicochemical properties, could be used as integral biosensor whose properties depend on temperature, the presence of chemical or biologically active compounds of different nature, etc. PMID- 12723479 TI - [The mechanism of specific cleavage of supercoiled DNA by human DNA topoisomerase I: the effect of ligand structure on the catalytic step of reaction]. AB - Eukaryotic DNA topoisomerase I (Topo) regulates the topological state of cell DNA and plays an important part in replication, transcription, repair, and recombination. Factors affecting the specific recognition of topologically stressed DNA were analyzed on the basis of the thermodynamic and kinetic data on the Topo-DNA interaction and the X-ray data on human Topo. A model was advanced for possible structural changes occurring in the ligand after initial recognition. The effect of conformational changes in specific DNA on the catalytic stage of the reaction was analyzed. PMID- 12723483 TI - [Development of the physician-patient relationship]. PMID- 12723481 TI - [Bifunctional reporter genes: construction and expression in prokaryotic and eukaryotic cells]. AB - Bifunctional reporter proteins were constructed to combine Clostridium thermocellum lichenase (LicBM2) with Aequorea victoria green fluorescent protein (GFP) or with Escherichia coli beta-glucuronidase (GUS). The major properties of the initial proteins were preserved in the hybrid ones: LicBM2 was active at 65 degrees C, GFP fluoresced, and GUS hydrolyzed its substrates. LicBM2 remained active after extension of its C of N end. Bifunctional reporter systems were shown to provide a convenient tool for studying the gene expression regulation in prokaryotic (E. coli) and eukaryotic (Saccharomyces cerevisiae, mammalian) cells, advantages of one reporter compensating for drawbacks of the other. PMID- 12723485 TI - [A simple audiometry test of speech intelligibility in background noise]. AB - We propose a new audiometric test to assess French speakers' speech discrimination in noise. This soundfield test uses a standard cocktail-party background noise superimposed on Fournier's disyllabic word lists. The background noise is CD-recorded at constant 55 dB SPL. The discrimination score is measured at a signal-to-noise ratio of -5 dB, 0, and dB. In comparison with other audiometric tests in noise, our test requires only 5 minutes and is simple for both patient and audiologist. All patients are tested using the same standard condition, one that approaches daily life and uses a noise of moderate intensity. The result is reproducible and easy to calculate and graph. We propose this test for diagnosing in-noise speech intelligibility disorders and for assisting the process of conventional or implantable hearing-aid fitting. PMID- 12723484 TI - [Utility and limitations of speech audiometry testing in the detection of occupational noise-induced hearing loss: a comparative study in two groups in the hospital setting]. AB - OBJECTIVE: The objective of this study is the evaluation of the calculation of 100% intelligibility threshold and noise resistance index as a monitoring test in populations exposed to noise in their work environment. METHOD: This is a retrospective exposed-unexposed study as the exposure to the risk factors took place before the survey. The exposed subjects were chosen in principle on sonometric criteria and the unexposed subjects among employees whom we believe were not exposed. The exploration method included history taking, clinical examination, free field audiometry and speech audiometry with background noise. RESULTS: The exposed population has a 100% intelligibility threshold on average equal to 29.69 dB, significantly higher than in the unexposed population where it is calculated at 25.30 dB (p = 0.07). The noise resistance index varies with age (p = 0.04). CONCLUSION: Our study shows the existence of a significant hearing difference between two populations with different noise exposure. It demonstrates a relationship between a hearing loss risk factor such as noise and the occurrence of the "professional hearing loss" disorder. The calculation of the 100% intelligibility index and the noise resistance index advantageously complements tonal audiometry. These procedures provide a better approach to the social impact of hearing loss but were not useful in the detection of hearing loss. PMID- 12723486 TI - [Necrotizing external otitis in children in Abidjan (Ivory Coast)]. AB - Necrotizing external otitis (NEO) is a fulminant Pseudomonas infection of the external auditory canal affecting mainly elderly diabetic patients. Since the early descriptions, many authors have related cases of NEO in non diabetic patients. We report eight cases of NEO in young children. They are less than two years old, they are undernourished and non diabetics. We get a good evolution in spite of our modest therapeutic ways. Emphasis is placed on efficiency of local remedy with colistine. PMID- 12723487 TI - Changing patterns of buccal manifestations in AIDS. AB - INTRODUCTION: Since the new antiviral drugs, e.g. protease inhibitors, arrived for the treatment of HIV-infected patients, the main oral infections associated with HIV disease have been brought under wider control. MATERIAL AND METHOD: We examined 214 HIV-1 infected patients, that were in-patients or presented for consultation at the ENT department of the Hospital das Clinicas (Sao Paulo, University Medical School), between January 1996 and November 1998. We review the different disorders which may affect the buccal cavity of HIV patients, 57 patients (26.6%). We divided the patients into two groups to compare the differences in appearance of oral lesions in those ones receiving two or three antiviral drugs. We had 53 patients presenting with oral lesions, the majority of them from the group receiving two drugs. RESULTS: The "P" value test was used and we concluded that there was a significant correlation between the use of triple antiviral therapy and a decrease of buccal lesions in HIV-infected patients. PMID- 12723489 TI - [Our experience with papillary microcarcinoma of the thyroid]. AB - INTRODUCTION: Papillary microcarcinoma (PMC) of the thyroid gland is high incidence pathology. Most of cases are related to a good prognosis. Conversely, numerous cases are reported with an aggressive course. Prognosis factors are age, size of tumour, multifocality, capsular invasion and cervical lymphadenopathy. MATERIAL AND METHOD: We have conduced a retrospective study about 4,208 consecutive thyroidectomies over a period of 16 years. 251 cases of papillary cancers were analysed including 64 PMC (25.5%). RESULTS: 90% of PMC were incidentally found and 59% were diagnosed during surgical procedure. Multifocality improved the sensibility of histological diagnosis (P = 0.04). In 78%, total thyroidectomy was performed associated with a selective neck dissection in 47%. In case of extemporaneous diagnosis all partial thyroidectomy was convert into total thyroidectomy. In case of delayed diagnosis and in the absence of unfavourable prognosis factor, no other surgical procedure was carried out. A 7 years mean follow-up revealed only one case of recurrence successfully treated with Iodine 131 ablation. There was no case of death due to PMC. CONCLUSION: This study confirms the good prognosis of PMC treated according to usual criterions. PMID- 12723488 TI - [Thyroid oncocytomas]. AB - OBJECTIVE: Thyroid oncocytomas are rare tumors. They put problems of differential diagnosis enter malignant and benign forms. The modalities of the surgical treatment are controversial, in form as benign as malignant. Their prognosis is also debated. MATERIAL AND METHOD: Our retrospective study concerns 111 thyroid oncocytomas, so 6.5% of thyroid operated in our centre between 1981 and 2001. The sex-ratio of the patients was 0.16 with an average age of 41 years (15 to 72 years). RESULTS: All the fixed nodules, those associated to a recurrential paralysis, to tangible nodes or/and to microcalcifications on the radiography of the neck was malignant. Histological extemporaneous exam was not decisive in 16% of cases, among which 33% showed themselves malignant in the definitive exam, which put in evidence 9 carcinomas (8%). We treated benign oncocytomas by a loboisthmectomy for the isolated nodules, and the subtotal or total thyroidectomy for multinodular glands. For oncocytic carcinomas, we realized a total thyroidectomy with ganglionic taking and histological extemporaneous exam, followed by evidment in case of invasion (2 patients). For the 5 patients having presented a residual fixation, we obtained a white cartography after radioactive iodine administration. For the benign oncocytoma, we did ot notice any recurrences with an average drop of 28 months (2 months to 7 years). For carcinoma, 8 patients on 9 presented an actuarial survival without disease with an average drop of 44 months (18 months to 8 years). CONCLUSION: The diagnosis of malignancy of thyroid oncocytomas can be strongly evoked on however fickle clinical and radiological criteria; it can be eliminated only after definitive anatomo-pathological exam. PMID- 12723490 TI - [Thyroid pseudo-nodules: 3 case reports]. AB - The thyroid pseudo-nodules constitute cervical lesions that can simulating neoplastic thyroid lesions either by their size or by their "cold" character in scintigraphy. We describe 3 cases of thyroid pseudo-nodules collected in 10 years: a thyroid hydatid cyst, a schwannoma of the recurrent nerve and a thyroglossal duct cyst. Throw these observations, the authors discuss the problems of differential diagnosis set out with these pseudo-nodules. Although the diagnosis in the 3 cases, was histological after post operative exam, the authors insist throw a review of the literature, on the place of fine needle aspiration cytology in the therapeutic strategy that can show specific signs for some lesions. For the other cases, surgery resection is effected to eliminate malignant lesions who are the principal differential diagnosis of thyroid pseudo nodules. PMID- 12723491 TI - [Carotid body paraganglioma: an uncommon localization. A case report]. AB - Paraganglioma are rare lesions, which come from neural cells. They arise the most frequently from carotid body. The tumor is classically located in the carotid bifurcation, between internal and external carotid arteries. We report a case of carotid body paraganglioma which is uncommonly located under the carotid bifurcation. PMID- 12723492 TI - [A rare tumor of the parapharyngeal space: myxoid chondrosarcoma]. AB - Extrasqueletal myxoid chondrosarcoma (EMC) is an uncommon soft tissue malignant tumor, locally aggressive with a high incidence of distant metastasis. It has distinctive clinical, immunophenotypic, cytogenetic and ultrastructural features. Most EMC are associated with the translocation t(9;22) (q22;q12). Their occurrence in the parapharyngeal space is extremely rare. Our objective is to discuss the difficulties of the histological diagnosis of EMC and to describe its immunophenotypic, cytogenetic features and clinical behavior. We report a case of a 67 years old woman who presented with a five months history of dysphagia. The oral examination found a mass displacing the posterior and left walls of the pharynx. Surgical resection of the tumor was undertaken. The pathologic examination concluded to the diagnosis of an EMC of the left parapharygeal space. Now, the patient is receiving an adjuvant radiotherapy. PMID- 12723493 TI - Schwannoma of the tongue: one case report. AB - The schwannomas are nervous tissue tumours that arise from Schwann cells; they are uncommon in peripheral nerves and rare in the tongue. After, a review of the literature, we present a case of schwannoma which arose in the ventral part of the body of the tongue. The tumour presented as a slowly growing mass producing few symptoms. The diagnosis of schwannoma is usually made post-operatively by histological identification although modern imaging techniques can provide useful indications. The treatment is exclusively surgical and usually enucleation of the mass is uncomplicated. There are no reports of malignant transformation. PMID- 12723494 TI - An important procedure in ossiculoplasty: autoclaving the ossicles. AB - Autoclaving of the ossicles prior to ossiculoplasty is a very important procedure in surgery of cholesteatomatous chronic otitis media. Autoclaving allows the reuse of the ossicles removed from patients with cholesteatomatous chronic otitis media as an autograft. It also allows utilization of the malleus, incus and stapes taken from cadavers or of the uneroded malleus and incus removed from patients undergoing non-functional middle ear surgery chosen carefully with detailed history and laboratory analysis. The powerful disinfecting effect of the sterilising procedures of the homografts inactivates prion proteins which cause degenerative encephalopathies. In various studies, it has been concluded that autoclaving does not alter the matrix of the bone which is responsiable for its biophysical properties, whereas it removes all viable cells within the bone and dentures the soft tissue attached to the surface of the ossicle. We have also found confirming histological results in our two previous studies published in 1999 and 2001. It is usually recommended that soft tissue on the surface of the ossicles is removed before autoclaving. It is also sufficient to autoclave the ossicles at 134 degrees C, and at 2.5 atmosphere pressure for 5 minutes in a flash autoclave. The autoclaving time of the homograft ossicles must be longer, for 20 minutes. Following the autoclaving, these homograft ossicles should be kept at pH 5.6 for 3 days, then use a solution of 0.5% formaldehyde at pH 7 and 4 degrees C for 21 days and it should be washed with physiologic saline solution three times for 7 minutes before use. Depending on the results obtained from our planned comparative experimental study, there will be no need for additional formaldehyde fixation procedure after autoclaving and the autoclaved ossicles will be used immediately without preservation in formaldehyde solution for 24 days. PMID- 12723495 TI - [Circadian rhythm and renal colic]. AB - Many physiological variables show a temporal pattern characterized by rhythmic changes along a time period, e.g. a day, a week, a month, a year, and it is also known that several acute diseases show a rhythmic variability as well. In these last years, high risk periods have been so identified, probably secondary to the temporal overlapping of favouring or triggering mechanisms. Recently, a circadian variability has been reported also for the occurrence of renal colic, with a pattern characterized by a morning peak, independent of gender and presence or not of demonstrable kidney stones. Most, if not all, renal functions exhibit temporal changes e.g. glomerular filtration rate, urine production and renal excretion of solutes, leading to an increased night-time concentration of urine, which could act as a predisposing factor for the morning occurrence of renal colic attacks. PMID- 12723496 TI - [The endocannabinoid system as a target for the development of new drugs for cancer therapy]. AB - Studies on the main bioactive components of Cannabis sativa, the cannabinoids, and particularly delta 9-tetrahydrocannabinol (THC), led to the discovery of a new endogenous signalling system that controls several physiological and pathological conditions: the endocannabinoid system. This comprises the cannabinlid receptors, their endogenous agonists--the endocannabinoids--and proteins for endocannabinoid biosynthesis and inactivation. Recently, evidence has accumulated indicating that stimulation of cannabinoid receptors by either THC or the endocannabinoids influence the intracellular events controlling the proliferation and apoptosis of numerous types of cancer cells, thereby leading to anti-tumour effects both in vitro and in vivo. This evidence is reviewed here and suggests that future anti-cancer therapy might be developed from our knowledge of how the endocannabinoid system controls the growth and metastasis of malignant cells. PMID- 12723497 TI - [Clinical policy for management and risk stratification of community-acquired pneumonia in patients hospitalized on the basis of conventional admission criteria]. AB - To identify discrepancies between Pneumonia Severity Index (PSI) risk class and the conventional criteria for deciding the site of care we performed a prospective observational study on 229 patients hospitalized for community acquired pneumonia. PSI classes and corresponding mortality rates were as following: class I, 41 patients (0%); class II, 20 (0%); class III, 58 (1.7%); class IV, 86 (8.1%); class V, 24 (33.3%). Overall, 119 patients (52%) who were hospitalized according to conventional criteria were assigned to low-risk classes (I-III). Among these low risk patients, 58 (49%) had complications as respiratory failure, pleural effusion, hypotension or shock; among remaining patients, no reasons for admission were found. This latter group deserves prospective evaluation in randomized studies comparing in-hospital versus outpatient management. PMID- 12723498 TI - [A multidisciplinary organizational Emergency Department model for trauma patients in hypovolemic shock]. AB - Hypovolemic shock in trauma patients causes high mortality because of exanguination, secondary brain damage, multiple organ disfunction. Good organization and operative protocols are vital to patients in need of a prompt and multidisciplinary treatment. PATIENTS AND METHODS: We collected data about resuscitative and diagnostic process, severity scores and outcome of all trauma patients in hypovolemic shock admitted to the Ospedale Maggiore in Bologna (Italy) after the organization of an Internal Trauma System. RESULTS: We obtained a reduction of both diagnostic and therapeutic times and mortality during the last 14 years. CONCLUSIONS: A valid internal organization can improve mortality of trauma patients without expenditure of extra resources. PMID- 12723500 TI - [Clinical surveillance of osteoporotic patients]. AB - The diagnosis of osteoporosis is often not considered, while many different pathophysiological conditions, as well as diseases, can determine or at least predispose to this syndrome. A detailed surveillance as well as an appropriate consideration of the general state of health and of the clinical history of the patient are therefore mandatory. Once established the diagnosis of osteoporosis, precise therapeutical directions are necessary as well as preventive strategies for evaluation of the disease and for the damages that can be derive: the fractures. It is a must to follow defined habits regarding life's style (i.e. appropriate diet, physical exercise, sun exposure, abstinence from drugs and excess of alcohol). Moreover, considering the age, the situation of the single patient as well as the results of specific tests (i.e.: densitometry, X-ray, biochemical markers of bone metabolism), the first step is whether it is appropriate to start a therapy with antiresorptive drugs. It takes a long lasting therapeutical intervention, either for years or sometimes for the entire life, in order to avoid fractures due to osteoporosis. These events are to be considered as the most dangerous consequences of osteoporosis, since they can worsen life's quality and cause a great burden of personal sufferance and of economical costs for the community. During these long lasting pharmacological interventions, the patients has to be checked at the appropriate time (usually every 18 months in terms of DXA), and at the most informative skeletal sites, in general the hip and the lumbar spine. Osteoporosis is a severe and chronic disease, and the related fractures impose an accurate consideration and surveillance by physicians. The patient has to be motivated by clear information of the risks related to disease, in order to obtain a good compliance as regards the above mentioned therapy and life's style habits. PMID- 12723499 TI - [Type I autoimmune hepatitis. An atypical case]. AB - Typically, autoimmune hepatitis (AIH) arises in young women, but recent papers have shown that it can affect older people too. The authors describe a case of AIH in a 70 ys old man affected by acute cholostatic hepatitis. The diagnosis of type I AIH was based on immunological features (high levels of ANA) together with histological picture in the absence of other known causes of hepatitis. Corticosteroid therapy is an effective treatment of type I AIH in older patients. AIH should be considered for the diagnosis of hepatitis in older male patients too. PMID- 12723501 TI - [Chronic obstructive lung disease. Systemic manifestations]. AB - Chronic obstructive lung diseases (COPD) are a complex disease state which not rarely can be associated with significant systemic manifestations. These alterations, though recognized since long time, are currently under extensive research, due to the increasing appreciation of their relevant negative role in the prognosis and health-related quality of life (Hr-QoL) of the COPD patients. The most clinically important are the decrease in body weight with loss of skeletal muscle mass (cachexia), osteoporosis, hypercapnia-induced peripheral edema, neuro-psychiatric disorders, such as oxygen-related cognitive impairment and depression, excessive polycytaemia and sleep disorders. Chronic systemic inflammation, oxidative stress and chronic hypoxia are believed as the main factors involved in the pathogenesis of systemic effects seen in COPD. Their adequate control with nutritional support, change of life-style and targeted pharmacological treatment is able to improve the prognosis and Hr-QoL among these COPD patients. PMID- 12723503 TI - [Image of the month. Isolated single coronary artery or blockage of the common arterial trunk and branches? ]. PMID- 12723502 TI - [Interstitial lung diseases]. AB - Interstitial lung diseases (ILD) are an heterogeneous group of inflammatory diseases characterized by an anatomical distortion of peripheral airways and interstitium, determined by a first stage of alveolitis and a following stage of fibrosis. Natural history of several ILD is characterized by slow and progressive destruction of alveolar-capillary functional units, often with respiratory failure and death. For their smoldering evolution and not specificity of symptoms (exertional dyspnea and cough) ILD may remain not diagnosed and not treated for a long time. PMID- 12723505 TI - [Clinical case of the month. Pulmonary toxicity due to gemcitabine for NSCLC with brain metastasis]. AB - Gemcitabine pulmonary toxicity is rare and represents a difficult diagnosis. A 61 year old female treated with gemcitabine for a metastatic non-small cell lung cancer (NSCLC) developed during the fifth chemotherapy cycle an acute respiratory distress syndrome with fever, tachypnea, marked hypoxemia and a diffuse interstitial-alveolar infiltrate on chest radiograph. No infectious or opportunistic etiology or cardiovascular disease was demonstrated. Withdrawal of gemcitabine and administration of corticosteroids led to symptomatic improvement. This acute pneumonitis was likely drug induced. PMID- 12723504 TI - [How I treat...severe hypoglycemia in a diabetic patient]. AB - Severe hypoglycaemic episodes are defined as need of assistance and may progress to profound coma. They can occur in patients treated with insulin, generally for type 1 diabetes, or in patients receiving sulphonylureas, for type 2 diabetes. Diagnosis is usually obvious, at least in insulin-treated patients, and requires an urgent intervention from the entourage. Such an intervention should comprise the oral administration of carbohydrates with high-glycaemic index if consciousness allows it or, if not, the injection of glucagon. When necessary, people should ask the help of a physician who will inject hypertonic glucose intravenously. Hypoglycaemic coma related to an absolute or relative excess of insulin should, in most cases, be treated at home. In contrast, a hypoglycaemic coma due to a too high dosage of sulphonylurea always requests a hospitalisation in order to carefully supervise the patient and to provide a prolonged intravenous infusion of glucose. It is mandatory that family or entourage members of any diabetic patient at risk to develop severe hypoglycaemia receive a specific education in order to promptly apply the best treatment capable of a rapid and safe recovery from hypoglycaemic coma. PMID- 12723506 TI - [Ozone exposure and asthma]. AB - Ozone is a pollutant the production of which depends on weather conditions and car engine combustion. Numerous epidemiological studies have indicated that high ozone levels correlated with morbidity in asthma. Experimental studies have shown that exposure of healthy subjects and asthmatics to ozone levels comparable to those measured in ambient air during hot summer days can generate respiratory symptoms, neutrophilic airways inflammation and lung function impairment. Lung function changes following ozone exposure are more pronounced in asthmatics and are dependent on the duration and intensity of exposure, a previous exposure and the nutritional status of the subjects. The airway epithelial cell layer is likely to play a pivotal role in initiating the inflammatory process following ozone exposure. Control of ambient air ozone levels must be a target for public health authorities. PMID- 12723507 TI - [Optimizing the management of type 2 diabetics: results of the "DREAM" study in general practice]. AB - This paper reports the results of the "DREAM" ("Diabetes REinforcement of Adequate Management") study performed among 42 general practitioners. A treatment algorithm was proposed to intensify the management of type 2 diabetic patients in case of poor control (HbA1C > 7.5% and fasting blood glucose > 140 mg/dl) before asking for an advice from a diabetologist. A total of 163 patients participated in the study: 84 men and 79 women, 59 +/- 10 years old; 5.2 +/- 6.1 years of known duration of diabetes; BMI: 30.7 +/- 5.8 kg/m2. Almost two thirds of patients received an antihypertensive agent, one third a lipid-lowering drug, and three quarters at least one antidiabetic agent: 22% a sulfonylurea, 18% metformin, 25% a combined oral therapy, 10% insulin (alone or in combination). At baseline, mean HbA1C averaged 7.1 +/- 2.2% (N: 4-6%), and 34% of patients had HbA1C > 7.5%. From these 163 patients, 100 were evaluated after 12 months, 57% being well controlled vs 36% at baseline. Among the 32 patients whose hypoglycaemic treatment was intensified, HbA1C level significantly decreased by almost 0.8% (p < 0.05). Despite precise guidelines, 55% of patients with HbA1C > 7.5% had no treatment adjustment, essentially in the group already on oral bitherapy or on insulin. Among the 65 patients evaluated after 24 months, the percentage of well controlled patients decreased to 44% (vs 51% after 12 months). The decision of switching to insulin was often delayed (no patient after 1 year and 2 patients only after 2 years). In conclusion, the "DREAM" study demonstrates the feasibility, but also the difficulty, of optimising the management of type 2 diabetic patients in general practice. PMID- 12723508 TI - [Nonalcoholic steatohepatitis: a review]. AB - The nonalcoholic steatohepatitis (NASH) is a liver disease of people who do not drink alcohol, with the histology of an alcoholic liver disease. It is the second or third most prevalent liver disease in western countries. The risk factors are the female gender, a middle age, a diabetes and a dyslipidemia. Some cases of NASH can reach the cirrhosis stage. The most common sign of the disease is an aminotransferase level elevation. PMID- 12723509 TI - [Alteration of bone metabolism in HIV-infected patients treated by HAART]. AB - For several years already, a growing number of studies reports modifications in the bone metabolism among HIV-infected patients. Some of these studies, published even before the use of HAART, involved the infection itself. With the experience already available as concerns HAART, antiretroviral treatments (ART) seem however to be called into question. Data are divergent yet. Some studies tend to invalidate the collected data about the harmful role of HAART and prove the absence of effect or even the beneficial action of ART on bone. Moreover, the three important classes of ART are implied, even if the proteases inhibitors are most commonly charged. Pathogenic mechanism remain hypothetical. While the impact on morbidity seems to be weak for the time being, long-term repercussions are still unknown, in particular when children are concerned. In such conditions, it appears difficult to set up coherent politics of screening, prevention and treatment. Nevertheless beyond the divergences, the multifactorial character of alteration of HIV-infected patient's bone metabolism seems to be undeniable. The identification of the different parameters should in the future clarify the situation and enable the publishing of exact criteria of screening, prevention and treatment. PMID- 12723510 TI - [How I explore...glycemic kinetics by continuous glucose monitoring]. AB - The improvement of diabetic patients' glycaemic status requires an increase of fingerstick blood glucose measurements. Among the possibilities available to assess diabetes' control, we will consider new systems of continuous interstitial glucose monitoring. After a brief description of the devices presently commercialized, we will discuss their indications and their limitations, as well as their future prospects in a possible "closed loop" insulin delivery according to blood glucose level. PMID- 12723511 TI - [Medication of the month. Topical pimecrolimus (Elidel)]. AB - Pimecrolimus inhibits some specific steps of the Th1 and Th2 immune reactions. It belongs to the class of ascomycin macrolactams. The topical drug formulation targets cutaneous inflammation, more particularly lymphocytes and mast cells without impairing systemic immunosurveillance. Topical applications are indicated for treating atopic dermatitis. In this indication, it represents an alternative or an adjuvant drug to topical corticosteroids. The treatment of other dermatoses could also benefit from pimecrolimus. PMID- 12723512 TI - [On-line monitoring of hemodialysis]. PMID- 12723513 TI - [Cardioprotection: an essential component for predialysis chronic renal failure treatment]. AB - Cardiovascular (CV) disease in uremic patients is a major concern to the nephrologist because it represents the main cause of morbidity and mortality in chronic renal failure patients, both predialysis and while on dialysis therapy. CV mortality is 3 to 20 times higher in dialysis patients than in the general population at similar age. Of note, a high prevalence of CV comorbidity is already present at start of maintenance dialysis, and is predictive of subsequent mortality on dialysis. CV disease progresses over years prior to the onset of ESRD, because risk factors develop from the early stage of chronic renal insufficiency. However, CV disease may be prevented or attenuated in patients who benefit from early, regular care of CV risk factors. Mechanisms of uremic cardiopathy, the major cause of mortality in uremic patients, are multifactorial and their effects are cumulative. Risk factors for left ventricular hypertrophy are hypertension, anemia, fluid overload and arteriosclosis, all of which are amendable by therapy. Risk factors for accelerated atherosclerosis, responsible for ischemic cardiopathy and myocardial infarction, are both common factors (e.g., hypertension, tobacco smoking and diabetes) and factors more specific for the uremic state (e.g., dyslipidemia, hyperhomocysteinemia and oxidative stress), all of which also are amendable by proper therapy. As a result, mixed hypertensive and ischemic cardiomyopathy develops, ultimately leading to cardiac failure, together with accidents resulting from valvular and arterial calcifications (favored by calcium-phosphate disorders), and from occlusion of coronary, cerebral and peripheral arteries. Cardioprotective therapy thus has become a cornerstone in the management of chronic renal failure patients, in conjunction with renoprotective therapy. Cardioprotective strategy involves optimal treatment of hypertension, anemia, fluid overload, dyslipidemia, hyperhomocysteinemia and calcium-phosphate disorders, and smoking cessation. To achieve a maximal efficacy, such treatment has to be initiated as early as possible in the course of renal failure. Because of its complexity, the integrated combined nephrotective and cardioprotective therapy requires early and sustained guidance by a nephrologist throughout the whole predialysis period. PMID- 12723514 TI - [Is there a place for ribavirin in the treatment for renal transplant patients infected by hepatitis C virus?]. AB - Treatment of chronic hepatitis C in renal-transplant (RT) recipients with alpha interferon is associated with a high rate of acute rejection. We therefore evaluated the biochemical, virological, histological efficacies, as well as the safety of one year ribavirin monotherapy in 16 HCV-(+) RNA (+) RT patients (group A) matched to 32 HCV-(+) RNA (+) RT patients (group B) who did not receive ribavirin. Ribavirin was initially started at a daily dose of 1000 mg and then adapted to hemoglobin level. Ribavirin monotherapy was associated with a significant decrease in AST, ALT and gamma glutamyl transpeptidase levels. Serum creatinine decreased as well. When proteinuria was present (n = 5), this decreased or disappeared. There was no significant changes in HCV viremia. The histological analysis of liver biopsies revealed a significant progression in liver fibrosis with no improvement in inflammation scores. There was a significant decrease in hemoglobin levels, despite an important support by recombinant erythropoeitin. However, in three cases, ribavirin therapy had to be stopped. In group B, after 1 year of follow up, there was a significant increase in serum ALT and creatinine values. Proteinuria decreased in only 2 of 12 patients. In conclusion, one year ribavirin therapy in HCV-(+) RNA (+)ve RT has no impact upon liver histology, although it improves liver enzyme levels. It impact upon renal function remains unknown. Nevertheless when proteinuria is present it disappears. PMID- 12723516 TI - [Value of isotretinoin in the treatment of acne in hemodialysis patients: apropos of 1 case]. AB - Rare are reports about isoretinoin treatment for acne in dialysis or kidney transplant patients whereas its efficacy and safety make it very interesting. Authors report here an additional observation of a 32 year-old young woman, hemodialyzed after one period of 12 years renal transplantation, and presenting a diffuse and severe acne. Treatment by isotretinoin with a moderate amount of 20 mg (0.38 mg/kg) per day cures this dermatosis in 2 months and half, without severe clinical side effect or significant metabolic disturbance, apart from a transient increase in parathormone rate. PMID- 12723517 TI - [Fast 1H-NR spectroscopic imaging of the human brain]. AB - Techniques allowing the reduction of the measurement time of MR spectroscopic imaging of protons (1H SI) were studied in a 1.5-T whole-body MR tomograph. The purpose was to facilitate the inclusion of the modality into an MRT routine protocol for examination of the human brain. The techniques were evaluated in a study with 6 healthy volunteers. Shortening of the measurement times can be achieved by reducing the number of phase-encoding steps with spherical k-space sampling, or by using delay times during TR periods for excitation and detection of several slices or echos. Since the tomograph enables echo-planar imaging, multi-gradiennt-echo techniques were implemented, thus sparing one phase-encoding gradient and shortening the measurement time by a factor of N. Due to signal loss, however, this technique is applicable only when the concentration of the metabolites of interest is sufficiently high to obtain a signal-to-noise ratio of at least 2 with a single acquisition. PMID- 12723515 TI - [Obstructive acute renal failure revealing visceral leishmaniasis in a diabetic patient]. AB - We report a case of a 22 years old type 1 diabetic man with a history of weight loss, weakness, anorexia, fever and recurrent urinary tract infection since February 2001. In April 2001, he presented anuria due to obstructive acute renal failure. Hepatosplenomegaly and lymphadenopathy were absent at physical examination. Laboratory tests revealed a high level of gamma globulin (53.4 g/l) and anaemia (haemoglobin 7.7 g/100 ml) without leukopenia and thrombocytopenia. CT scan showed multiple retroperitoneal lymphadenopathies causing compression of the two ureters, hydro-ureter associated with hydronephrosis, hepatosplenomegaly and multiple pulmonary nodes. Lymphadenopathies, anaemia, high level of gamma globulin, high titres of anti-leishmanial antibodies and the excellent outcome after treatment with meglumine antimoniate (Glucantime) confirmed visceral leishmaniasis. This report documented an unusual clinical presentation of Visceral leishmaniasis in a diabetic patient. PMID- 12723518 TI - [Fluence-modulated radiotherapy with an optimization-integrated sequencer]. AB - On the basis of two clinical cases, we present fluence-modulated radiotherapy with a sequencer integrated into the optimization of our treatment-planning software HYPERION. In each case, we achieved simple relations for the dependence of the total number of segments on the complexity of the sequencing, as well as for the dependence of the dose-distribution quality on the number of segments. For both clinical cases, it was possible to obtain treatment plans that complied with the clinical demands on dose distribution and number of segments. Also, compared to the widespread concept of equidistant steps, our method of sequencing with fluence steps of variable size led to a significant reduction of the number of segments, while maintaining the quality of the dose distribution. Our findings substantiate the value of the integration of the sequencer into the optimization for the clinical efficiency of IMRT. PMID- 12723519 TI - [Radiotherapy treatment planning for small complex-shaped lesions and its experimental verification]. AB - The Gamma Knife is used as a stereotactic tool for the conformal treatment of very small, complex-shape cranial lesions. The combination of planning software and treatment equipment enables a highly-precise conformal dose distribution and positioning. The purpose of the present study was to experimentally verify the precision actually achievable in case of extremely irregular, small target volumes. For this purpose, a complete treatment procedure was performed using a standard head phantom complemented with a specially developed insert that simulates an L-shaped lesion. The spatial precision of the irradiation was recorded by means of high-resolution film dosimetry using GafChromic films. The analysis of the films showed for the film in the center plane an excellent conformity of the 75% isodose line used to circumscribe the lesion. A very good agreement between planning and measurement resulted also for isodose lines residing outside of the target volume. PMID- 12723520 TI - [Improvement of the accuracy of the Laplace transform method for the determination of radiotherapy spectra of clinical linear accelerators]. AB - The present study focused on the reconstruction of the bremsstrahlung spectrum of a clinical linear accelerator from the measured transmission curve, with the aim of improving the accuracy of this method. The essence of the method was the analytic inverse Laplace transform of a parameter function fitted to the measured transmission curve. We tested known fitting functions, however they resulted in considerable fitting inaccuracy, leading to inaccuracies of the bremsstrahlung spectrum. In order to minimise the fitting errors, we employed a linear combination of n equations with 2n-1 parameters. The fitting errors are now considerably smaller. The measurement of the transmission function requires that the energy-dependent detector response is taken into account. We analysed the underlying physical context and developed a function that corrects for the energy dependent detector response. The factors of this function were experimentally determined or calculated from tabulated values. PMID- 12723521 TI - [Advanced method for calculating the scatter signal contribution in CT detectors by the Monte Carlo method]. AB - Multislice spiral CT scanners allow to acquire multiple slices simultaneously. With increasing numbers of slices, not only the total extent of slice collimation increases, but also the contribution of scatter radiation to the detector signal. A fast method for calculating the scatter signal would offer the possibility to correct the measured detector signal. Monte Carlo methods allow to simulate the paths of photons through a 3D volume, both in a patient- and scanner-specific fashion. If a scatter photon leaves the volume, its path can be followed and its interaction with an element of the detector be checked. This conventional way of calculating the scatter signal is time-consuming. In order to reduce the calculation time, a more efficient method was developed (Method of Weights). Every time an interaction occurs inside of the 3D volume, the probability of a detector hit due to photon scattering is calculated for each detector channel. The respective value is added to the scatter signal per detector with the corresponding weight. Simulated values of scatter-to-primary-signal ratios were confirmed by data available in the literature. Both the conventional and fast methods for the calculation of scatter signals yielded identical values within the range of statistical accuracy. Assuming the same computing time, the standard deviation for the conventional method was 5 times higher than for the fast one. The presented method allows to significantly reduce the computation time. It may therefore provide a basis for "real time" methods to correct for the scatter signal, especially in case of increasing numbers of slices. PMID- 12723522 TI - [Validation of constancy testing of film processing according to Regulation 6868 2]. AB - Constancy tests of film processing according to DIN 6868-2 require the measurement of the quantities "Empfindlichkeitsindex EI" (Speed Index) and "Kontrastindex KI" (Contrast Index), aimed at determining changes of film processing that can influence the resulting radiographic image. In this study it was investigated whether these quantities are actually suited to adequately describe processing-induced changes of the sensitometric properties of X-ray films. For this purpose, processing-induced changes of the parameters EI and KI were compared with corresponding changes of the parameters "Lichtempfindlichkeit LE" (Light Speed) and "Lichtkontrast LK" (Light Contrast), both used for acceptance tests of film processing according to DIN V 6868-55. The results of this study can be summarised as follows: 1. There was a very good correlation between the quantities EI and LE. 2. Changes of the quantity KI were generally not correlated with changes of LK, for example significant changes of LK were not always reflected by changes of the quantity KI. The following conclusions are drawn: 1. The quantity EI is well suited to indicate changes of the sensitivity of film processing. 2. The use of the quantity KI for constancy testing is questionable, and it is suggested to refrain from the measurement of KI. PMID- 12723523 TI - [R50 and Rp in electron beam dosimetry]. AB - The practical range Rp and the half-value depth R50 are relevant parameters in clinical dosimetry of electron beams. The aim of this study was to determine a relation between Rp and R50, as well as between Rp and the mean energy at the phantom surface Eo. A total of 135 depth-absorbed-dose curves from 20 different accelerators were analyzed. The relation between Rp and R50 could be fitted by a linear equation. This equation was compared with the equation calculated in the literature by means of Monte Carlo simulation. The prevailing data confirm that a linear relation exists between Rp and Eo. PMID- 12723524 TI - [Staffing in medical radiation physics in Germany--summary of a questionnaire]. AB - To obtain an overview of the actual staffing levels in Medical Radiation Physics, a survey was carried out in 1999 by the task-group "Staffing requirements" of the Deutsche Gesellschaft fur Medizinische Physik (DGMP; German Society of Medical Physics) among all DGMP members active in this field. The main components for equipment and activities are defined as in Report 8 and 10 of the DGMP for staffing requirements in Medical Radiation Physics. The survey focused on these main components. Of 322 forms sent out, 173 answers could be evaluated. From the answers regarding equipment and activities, theoretical staff requirements were calculated on the basis of this spot-check target and compared with the effective staffing levels documented in the survey. The spot-check data were then extrapolated to the whole Germany. The calculation revealed a deficit of 72% for the whole physics staff and of 58% for the number of physicists. Considering the age distribution of the DGMP members and the calculated staffing deficit, a training need was calculated of approximately 100 medical physicists per year in Germany, provided that the goal is set of cutting back the deficit in 10 years. PMID- 12723525 TI - [Radiotherapy and breast reconstruction: the issue of compatibility]. AB - BACKGROUND: Breast reconstruction after mastectomy represents an improvement in the quality of life of breast cancer patients. Radiotherapy is also suggested for many of them to prevent local relapse. However, irradiation increases the risk of complications require surgical procedures. PURPOSE: This study was undertaken to analyse the compatibility of breast reconstruction and irradiation in the treatment of breast cancer. METHODS: Computerised searches for publications debating this issue were done of MEDLINE data. Studies were grouped according to the techniques and types of reconstruction: reconstruction with silicone implant or autogenous skin-muscle flap, timing of reconstruction (immediate or delayed), and sequencing of treatments (pre- or post-reconstruction irradiation). The results of studies were assessed and compared in respect to reconstruction related chronic complications. RESULTS: The incidence of complications was significant even in the absence of irradiation, but radiotherapy increased the risk of complications to less or more extent. When reconstruction was done with implant, the most common type of complication was Grade III-IV capsular contracture. In patients subjected to immediate reconstruction, the complication rates with or without radiotherapy were from 0% to 64% and from 0% to 12%, respectively. In women who underwent delayed reconstruction, the incidence of complications with or without irradiation was from 22% to 55%, and from 17% to 34%, respectively. The negative effect of radiotherapy was more significant with immediate than with delayed reconstruction. In patients who underwent reconstruction with skin-muscle flaps, the most common type of complication was skin necrosis. The incidence of complications with or without radiotherapy was from 12% to 39% and from 5% to 25%, respectively. Cancer, developing after cosmetic augmentation mammaplasty in the breast, can be treated with lumpectomy and radiotherapy without removal of the implant. In series with this method of treatment the incidence of complications (mainly capsular contracture) was from 0% to 65%. The use of moderate dose (45-50 Gy), wedge filters, and no use of bolus application decreased the risk of complications. CONCLUSIONS: Radiotherapy and breast reconstruction are not incompatible, but careful consideration of their relative timing and technique is important. Plastic surgeons should counsel patients before starting their cancer disease treatment, and those who choose to have reconstruction need to be informed about risks for specific complications associated with the procedure. Results of the studies debating this issue are controversial. Longer follow-up time, larger patient material and better specified parameters are needed to validate results. PMID- 12723526 TI - [Lipid-lowering therapy in patients with type-2 diabetes mellitus]. AB - Diabetic dyslipidemia is mainly characterised by hypertriglyceridaemia, low HDL cholesterol level, an increased small dense HDL concentration, i.e. by atherogenic dyslipidemia. Dyslipidaemia occurs in some two third of the type 2 diabetes cases. In the treatment of dyslipidaemia it is essential to control the diabetes, to reduce the intake of saturated fat and supplement it with monounsatured fat ty acid or complex carbohydrates. Based on the latest studies diabetes is considered the same risk as coronary heart disease and, therefore, diabetic dyslipidaemia should be treated in the same aggressive way. According to the simplified guidelines, after the diet--above 5.2 mmol/l cholesterol level- antilipaemic drugs, i.e. statin should be administered in order to achieve the primary goal of the therapy, namely the 2.6 mmol/l LDL-cholesterol level. In patients with combined II/b type hyperlipoproteinaemia statins are the drugs of first choice, fibric acid derivates being only considered in case of normal LDL cholesterol level (< 3.4 mmol/l), if the HDL-cholesterol level is also low. Fibrate therapy is the first choice in the isolated hypertriglyceridaemia (> 2.3 mmol/l) as well as in type V. hyperlipoproteinaemia. On the basis of the guidelines far more patients with diabetes should be treated with lipid lowering therapy than before. PMID- 12723527 TI - [Optimizing estrogen treatment in Turner syndrome]. AB - INTRODUCTION AND AIM: Authors deducibled from retrospective analysis of patients data with Turner syndrome, who reached near adult height and from results of many multicentre study, that induction of puberty should be designed individually. They show, that estrogen therapy used in their earlier practice for growth promoting did not improve final height of patients. METHODS: The patients were assigned into three groups. The group 1. consisted of untreated patients, who had spontaneous puberty (n = 10). The group 2. consisted of patients, who received oxandrolone and estrogen (n = 18). The group 3. consisted of patients treated with growth hormone (n = 17). In those patients received growth hormone estrogen treatment was started at the age of 14.7 +/- 1.97 years. RESULTS: The final height or near adult height was 144.0 +/- 4.6 cm, 143.5 +/- 1.8 cm, and 154.2 +/- 7.0 cm in group 1, 2 and 3 respectively. The final height was not greater in the group 2. compared to that found in patients who developed spontaneous puberty (group 1.). CONCLUSION: The individual estrogen therapy in patients treated with growth hormone allows feminization, as well as the best adult height, without the occurrence of the more serious mental disturbances. PMID- 12723528 TI - [Comparative analysis in artificial nutrition]. AB - INTRODUCTION: It is well known to Hungarian experts in the field of nutrition that hospital inpatients in this country do not have the required nutrition. AIM: To compare data of intensive care national costing analysis from England and intensive care unit in County Hospital, Eger, Hungary in order to advise a cost effective nutrition protocol. METHOD: English data were extracted from the report of Intensive Care National Cost Block Programme, year 1999. The Hungarian data were obtained by top down method from annual costing report of the same year. The authors used Purchasing Power Parity to make international cost comparison between these countries. RESULTS: In proportion to Purchasing Power Parity, the hospital budget per patient for nutrition is more than double in England than in the studied intensive care unit in Hungary. Intensive care units in England spend 1% on nutrition, 13% on drugs and 10% on disposables. There is only 0.2% spent on nutrition, 29.8% on drugs and 8% on disposables in the studied intensive care unit in Hungary. Cost of nutrition per patient day is HUF 117 in Hungary, which is even lower than the hospital budget per patient day for nutrition. CONCLUSION: On the basis of the above findings and literature review as well, the authors set up a cost effective nutrition guideline: 1. Identify malnourished patient. 2. Nutrition can be delayed for 4-5 days in not malnourished patient. 3. Enteral nutrition should always be the first choice. 4. Total parenteral nutrition is beneficial only if given over at least 7 days. 5) It is cost effective to prevent nosocomial infections even at higher cost. PMID- 12723529 TI - ["Devotions upon emergent occasions": medical views of John Donne]. PMID- 12723530 TI - [Soma Weiss (1899-1942)]. PMID- 12723531 TI - [External comparative quality assurance in cholecystectomy. Results of the Federal evaluation, 2001]. PMID- 12723533 TI - [DRG exercises. No fear of errors: the G-DRG 901Z and 902Z]. PMID- 12723534 TI - [Stand-by service as work time: does everything remain the same after the decision of the Federal Labor Court?]. PMID- 12723536 TI - [Borrowed feathers. Shaman medicine on the positive list]. PMID- 12723535 TI - [Flexible work time for physicians. Experience with work sheets and a system of time accounting at the Alfried Krupp Hospital]. PMID- 12723537 TI - [Comment on the article by U. H. Hohmann: Nonsense of certification. Der Chirurg BDC (2002) 41:303]. PMID- 12723539 TI - [A large coalition for health care reform?]. PMID- 12723540 TI - [The cards are on the table]. PMID- 12723541 TI - [Postoperative pain therapy. Synopsis from Chirurg, issue 3/2003]. PMID- 12723542 TI - [Wilms tumors. Diagnosis and therapy]. PMID- 12723543 TI - Ethical challenges of palliative care research. PMID- 12723544 TI - [Off-label prescriptions. Drug use outside of established guidelines and treatment standard]. PMID- 12723545 TI - [Rare variations in exstrophy-epispadias complex]. PMID- 12723546 TI - [Will health insurance survive the year 2006?]. PMID- 12723547 TI - [The Dusseldorf Urology Network. An economic association as an alternative to the national health program organization]. PMID- 12723548 TI - [Requirements regarding safety information of vasectomy]. PMID- 12723549 TI - [AndroCHECK: "The Urology--a specialist for men" Project"--and now?]. PMID- 12723550 TI - [Hormone substitution with testosterone gel]. PMID- 12723552 TI - Analysis of continuing education and scientific activities at the 2002 WFNMB meeting, Santiago, Chile. PMID- 12723551 TI - Imaging of renal structure and function. Post-congress symposium to the EANM Congress 2002 in Vienna Ceske Budejovice, Czech Republic, 5-6 September 2002. PMID- 12723553 TI - Portugal gets its first full-ring high-resolution dedicated PET camera 18 October 2002, Porto, Portugal. PMID- 12723554 TI - Sensitivity or resolution - or both? "State of the Art Imaging Using Small Animal PET Scanners" report on the 10th Bottstein Colloquium/2nd Workshop on Basic Research in Molecular Imaging, 11-12 October 2002, Villigen, Switzerland. PMID- 12723555 TI - Summary of the Third International Sentinel Node Conference 16-18 November 2002, Yokohama. PMID- 12723556 TI - HTBS advises new PET facility in Scotland. PMID- 12723557 TI - EANM procedure guidelines for treatment of refractory metastatic bone pain. PMID- 12723558 TI - EANM procedure guidelines for radiosynovectomy. PMID- 12723559 TI - Addendum to Freud's "Criminals from a sense of guilt". PMID- 12723560 TI - EANM procedure guidelines for 32P therapy. PMID- 12723561 TI - Guidelines for 131I-ethiodised oil (Lipiodol) therapy. PMID- 12723562 TI - Guidelines for 131I-meta-iodobenzylguanidine therapy. PMID- 12723563 TI - EANM procedure guidelines for therapy with iodine-131. PMID- 12723564 TI - Recognizing and addressing the mental health needs of children: a new understanding of treatment options and efficacy. PMID- 12723565 TI - Epilepsy in England. PMID- 12723566 TI - Standards for privacy of individually identifiable health information. HIPAA implementation. PMID- 12723567 TI - [Michelle Georges-Smoular: break the isolation of occupational health nurses]. PMID- 12723568 TI - [Magnetic field therapy--effective treatment or nonsense?]. PMID- 12723569 TI - [SARS: a new paramyxovirus or coronavirus?]. PMID- 12723570 TI - Human Leucocyte Differentiation Antigen nomenclature: update on CD nomenclature. Report of IUIS/WHO Subcommittee. AB - The 7th International Workshop on Human Leucocyte Differentiation Antigens (HLDA7) studied a number of newly characterised molecules relevant to human leucocyte differentiation and function. The HLDA organisation, which devised and continues to maintain the CD nomenclature, is responsible, under the auspices of IUIS and WHO, for the nomenclature of all leucocyte differentiation markers. The 7th Workshop redefined a number of (principally carbohydrate) molecules, and assigned CD names to approximately 80 new molecules. This update lists, in tabular form, the redefined and newly assigned names, together with antibodies, which have been confirmed under Workshop conditions as specific for the new and redefined molecules. The major features of the cellular expression patterns are summarised, and a LocusLink accession number provided to enable the reader to access more detailed information through http://www.ncbi.nlm.nih.gov/LocusLink. PMID- 12723571 TI - [Consensus on basic principles of team work between gastroenterologists and visceral surgeons]. PMID- 12723573 TI - [Antibodies and oxidative stress]. PMID- 12723572 TI - [Correlation between dermoscopy and histopathology in pigmented and non-pigmented skin tumours]. AB - Dermoscopy (dermatoscopy, epiluminescence microscopy, surface microscopy) is a non-invasive method in dermatology. With this method pigmented and non-pigmented skin tumours can be diagnosed with a clear higher sensitivity and specificity compared to clinical examination. Malignant skin tumours are detected earlier and unnecessary excisions of benign skin tumours can be avoided. The knowledge about the correlation between dermoscopy and histopathology is hereby an essential condition. In this continuing medical education article the correlation of both examination techniques is presented for the dermoscopic differential diagnoses. PMID- 12723574 TI - [Vision disorders in therapy with fumaric acid esters?]. PMID- 12723575 TI - [Corneal abscess with the threat of perforation]. PMID- 12723576 TI - [Multiple tumors of the facial skin]. PMID- 12723577 TI - [Angiogenesis in ophthalmology. A theme with a future]. PMID- 12723578 TI - Prediction of improved vision in the amblyopic eye after visual loss in the non amblyopic eye, by J.S. Rahi, S. Logan, M. Cortina-Borja, C. Timms, I. Russel Eggit, and D. Taylor. Lancet 360: 597-602, 2002. PMID- 12723579 TI - Talaporfin sodium. LS 11, ME 2906, mono-L-aspartyl chlorine e6, NP e6, NPE 6, taporfin sodium. PMID- 12723580 TI - Medicare mental health coverage. AB - Medicare generally covers less than half the cost of mental health-care services. In order for you to best help your clients with mental health-care needs, it is critical to understand Medicare's mental health benefits and gaps. In this brief, we present an overview of mental health coverage and payment policies, including consumers' costs and their rights to appeal denials of payment. PMID- 12723581 TI - Prognostic significance of persistent mediastinal metastasis following induction therapy in large (> or = 2 cm) N2 or N3 non-small cell lung cancer. AB - OBJECTIVE: It is controversial whether or not surgery is beneficial for patients with non-small cell lung cancer accompanied by persistent lymph node metastasis in the mediastinum following induction therapy. We have therefore conducted a retrospective study to assess this issue. METHODS: Eligibility criteria were defined as follows: 1) the period of treatment was between January 1991 and April 1998, 2) the clinical stages were IIIA (N2) or IIB (N3) with large lymph nodes (> or = 2 cm), 3) induction therapy had been administered, 4) tumor was resected completely, 5) at least one mediastinal lymph node had necrosis or scar if the pathological N status was p-N0 or p-N1 and 6) the p-stage was not IV. Dichotomous variables included the radiographic response of the tumor, the T status, and the N status. RESULTS: Thirty-nine patients were eligible. There were 29 males and 10 females aged from 27 to 74 years, and involved 20 cases of adenocarcinoma. The pathological N status was as follows: p-N0 in 18 patients, p-N1 in 3, p-N2 in 16, and p-N3 in the other 2. In overall survival, the median survival time (MST) was 34 months and the actuarial 5-year-survival rate (5-YSR) was 28%. The group of patients with either N0 or N1 (n = 21) had a 71-month MST and a 54% 5-YSR, and the group of patients with either N2 or N3 (n = 18) had a 13-month MST and a 5 YSR of 0% (p < 0.0001). On multivariate analysis, the pathological N factor was confirmed as an independently significant. CONCLUSIONS: Our retrospective study found that the survival rate of patients with persistent mediastinal nodal metastasis was very poor. A prospective study is needed to investigate whether or not surgery is beneficial for these patients. PMID- 12723582 TI - Off-pump multivessel revascularization: efficacy of suction type of coronary stabilizer. AB - OBJECTIVE: Off-pump coronary artery bypass grafting (CABG) has come into widespread use with the availability adequate coronary stabilization devices. We studied the efficacy of second-generation coronary stabilization devices (suction device) comparing to the first-generation device (compression device). METHODS: We prospectively analyzed consecutive patients who underwent isolated off-pump CABG via a midline sternotomy at Shin-Tokyo Hospital Group between July 1, 1996, and August 31, 2000, comparing perioperative, and follow-up data in the group using a suction device (group S) to that in the group using a compression device (group C). RESULTS: Preoperative risk factors were identical between the two groups, with the exception of a higher incidence of three vessel disease in group S. Complete revascularization increased from 47.3% in group C to 88.1% in group S, and the number of distal anastomoses from 2.1 +/- 0.6 in group C to 2.9 +/- 0.9 in group S. Revascularization of the circumflex artery was achieved in 21.7% of group S patients, which was significantly higher than that in group C (2.2%). Postoperative recovery, mortality, and morbidity did not differ significantly between groups. Calculated event-free rates at 2 years was 88.7% in group C and 92.0% in group S (p = NS). CONCLUSIONS: Anastomosis to the posterior wall of the heart using the suction device is safe. An increased number of distal anastomoses may reduce the occurrence of cardiac events related to incomplete revascularization. PMID- 12723583 TI - Surgical treatment of traumatic thoracic aorta rupture: a 7-year experience. AB - OBJECTIVES: Traumatic aortic rupture is highly lethal and an ongoing therapeutic challenge. We review our 7-year experience with traumatic aortic disruption. METHODS: We treated 12 cases of traumatic rupture of the thoracic aorta (TRTA) from December 1994 to June 2001 at our institution. Of these, 9 were male, and the average age 26 years. Injuries were caused by traffic accidents in 9 cases and falls in 3. Contrast-enhanced helical computed tomography was used to diagnose10 cases and digital subtraction angiography to diagnose 2 at other hospitals. Six of 12 (50%) disruptions were located in the aortic isthms. All surgery was conducted under cardiopulmonary bypass. A percutaneous cardiopulmonary support system (heparin-bonded artificial lung and centrifugal pump) was used in 6 cases since 1998. RESULTS: Among the 12 patients, 6 had early surgical repair within 2 days after the accident, and all survived free of neurological problems. Six other had repair delayed more than 2 days and all were doing well. CONCLUSION: Immediate repair of aortic lesions should be the rule because the majority of deaths from TRTA occur within 24 hours. We believe, however, that immediate surgery may not be necessary for some patients with severe, multiple associated lesions who survive initial traumatic aortic disruption of the aorta. PMID- 12723584 TI - Myocardial ischemic preconditioning during minimally invasive direct coronary artery bypass grafting attenuates ischemia-induced electrophysiological changes in human ventricle. AB - OBJECTIVE: Ischemic preconditioning (IPC) has been found to protect the myocardium in animal studies. However, clinical studies have been limited and the clinical effects of IPC are still uncertain. The purpose of this study was to assess whether IPC has any protective effect on the human myocardium during minimally invasive CABG (MIDCAB), by means of epicardial electrophysiological testing. METHOD: Forty-five patients with left anterior descending artery disease who underwent a MIDCAB procedure were evaluated. In the present study, the electrical potentials which were not affected by cardio-pulmonary bypass or cardioplegia were measured. The ratio of longitudinal to transverse conduction velocity (phiL /phiT), and QT, JT dispersions were measured using plaque electrodes in the preischemic state, during a 5-minute coronary occlusion, during the subsequent 5-minute reperfusion, during 5- and 10-minute anastomosis periods, and after anastomosis. RESULT: The phiL/phiT was 2.2 +/- 0.2 at baseline. Anisotropy was exaggerated during the 5-minute coronary occlusion (2.6 +/- 0.3). During anastomosis, conduction velocities were decreased, but showed no further deterioration (2.4 +/- 0.3, and 2.4 +/- 0.3, respectively). QT and JT dispersions were improved by reperfusion. CONCLUSION: The effectiveness of IPC during the MIDCAB procedure was confirmed electrophysiologically. Anisotropy and dispersions were minimized after IPC, therefore IPC demonstrated antiarrhythmic protective effects on the human myocardium. PMID- 12723585 TI - Successful aortic valve replacement for severe aortic regurgitation combined with annular abscess due to active infected endocarditis: usefulness of gelatine resorcine-formol biological glue in remodeling the new aortic annulus. AB - Two men, 58 and 72 years old, were diagnosed as severe aortic regurgitation complicated by aortic annular abscess due to active endocarditis infection diagnosed by echocardiography. We conducted aortic valve replacement using gelatine-resorcine-formol biological glue to close the abscess cavity and remodel the new aortic annulus. Although 1 man developed complete atrial-ventral blockage postoperatively and required that a permanent pacemaker be implanted, neither experienced recurrence of infectious peri-valvular leakage. PMID- 12723586 TI - Real-time cerebral monitoring using multichannel near-infrared spectroscopy in total arch replacement. AB - Multichannel near-infrared spectroscopy (NIRS) could detect change in the regional cerebral oxygenation by following animated pictures of oxy-hemoglobin (OxyHb), deoxy-hemoglobin (DeoxyHb) and total hemoglobin in operations for three surgical cases of thoracic aortic aneurysm with selective cerebral perfusion (SCP). Simultaneously measured jugular venous oxygen saturation (SjO2) showed no change in parallel to OxyHb or DeoxyHb of NIRS. It was concluded that SjO2 represented the entire rather than the local findings of the cerebral oxidative metabolism. Assessment of the intra cranial oxidative metabolism using a multichannel NIRS provided real-time information about the efficacy of SCP, while SjO2 had a comprehensive limited value. The animation enabled the detection of regional hypoperfusion visually and instantly during SCP. This multichannel NIRS was a new real-time monitoring method and was useful to prevent cerebral neurological complication due to hypoperfusion during SCP. PMID- 12723587 TI - Herpes simplex viral pneumonia after coronary artery bypass grafting. AB - We report a case of herpes simplex viral (HSV) pneumonia as a post-CABG pulmonary complication in a 70-year-old man. Chest radiography on postoperative day 9, showed a glass-like shadow and pleural effusion in the left lung field, and the man's condition began deteriorating rapidly. Bronchofiberscopy to detect the pathogen and a bronchoalveolar lavage with polymerase chain reaction (PCR) yielded a definitive diagnosis of HSV pneumonia. Once therapy with acyclovir was begun, his condition improved markedly. Our case suggests that 1 viral pulmonary infection should be considered as a possible cause in postoperative cardiac patients with unexplained progressive pulmonary infiltrates, and 2 DNA amplification using PCR is rapid--it can be completed within 1 day--and sensitive and specific in diagnosing such infections. PMID- 12723588 TI - Surgical repair of pulmonary arterial sling associated with tetralogy of Fallot. AB - Pulmonary artery sling associated with tetralogy of Fallot was successfully repaired in 2 patients. In 1 patient, extensive reconstruction was needed for severe hypoplasia in the left pulmonary artery, followed by definitive repair. In the other patient, surgical repair was achieved in a single-stage fashion. PMID- 12723589 TI - Pulmonary resection with chest wall removal and reconstruction for invasive pulmonary mucormycosis during antileukemia chemotherapy. AB - A 54-year-old woman undergoing chemotherapy for acute myeloid leukemia developed invasive pulmonary mucormycosis in the right upper lobe at the neutropenic nadir. Amphotericin B therapy became ineffective after an abscess formed in the affected lung, and insufficient infection control compelled us to interrupt chemotherapy. The lesion was suspected of invading the anterior chest wall. After right upper lobectomy combined with the anterior chest wall resection, the chest wall defect was reconstructed using autologous free rib grafts. Successful control of the fungal infection by resection enabled us to restart chemotherapy with concomitant use of Amphotericin B. In selected cases of leukemia-associated pulmonary mucormycosis refractory to Amphotericin B therapy, aggressive surgical intervention may facilitate anti-leukemia chemotherapy and prolong survival. PMID- 12723590 TI - Chondrosarcoma of the rib. AB - A 73-year-old woman evidencing an abnormal shadow on chest X-ray film since 1993 was admitted after a tumor of the left 8th rib was diagnosed in 2001. Computed tomography showed a low-density mass with coarse calcification arising from the left 8th rib and protruding into the abdominal cavity. The tumor was diagnosed as low-grade chondrosarcoma by incision biopsy and was resected together with the left 7th, 8th, and 9th ribs. The chest wall was reconstructed using Marlex mesh. Histological findings were compatible with Grade I chondrosarcoma. The patient had a long clinical course without distant metastasis thanks to the tumor's low malignancy. PMID- 12723591 TI - Type II antifreeze protein from a mid-latitude freshwater fish, Japanese smelt (Hypomesus nipponensis). AB - A lot of reports of antifreeze protein (AFP) from fish have been published, but no report has mentioned of commercialized mid-latitude fresh water fish which producing AFP in its body fluid. We found that the AFP in the body fluid of Japanese smelt (Hypomesus nipponensis) from mid-latitude fresh water was purified and characterized. The N-terminal amino acid sequence of the Japanese smelt AFP was 75.0% identical to Type II AFP from herring. Results of EDTA treatment and ruthenium red staining suggested that the Japanese smelt AFP had at least one Ca2+-binding domain. Interestingly, the antifreeze activity of the Japanese smelt AFP did not completely disappear when Ca2+ ions were removed. The molecular mass of the Japanese smelt AFP was calculated to be 16,756.8 by the TOF-mass analysis. The Open reading flame of the gene coding for the Japanese smelt AFP was 444 bp long and was 85.0% identical with the entire herring AFP gene. The cDNA and amino acid sequence of the Japanese smelt AFP were the same length as those of herring AFP. PMID- 12723592 TI - Elimination of cell-cycle regulators during caspase-3-dependent apoptosis caused by an immunosuppressant, FTY720. AB - The immunosuppressant, FTY720 causes apoptosis of lymphocytes, reduces numbers of lymphocytes in peripheral blood, and prevents infiltration of lymphocytes into allografts, which may be one of the mechanisms involved in its effects. Here we compared caspase activation and expression of cell-cycle regulators during apoptosis caused by FTY720, and Fas-stimulation in a mouse lymphoma transfected with human Fas antigen. FTY720 activated caspases-3, -8, and -9 as rapidly as did Fas-mediated apoptosis. The activation was blocked by a peptide inhibitor for caspase-3, DEVD-CHO. Fas-induced activation of caspases-8 and -9 was unaffected by the inhibitor. FTY720 eliminated proliferating cell nuclear antigen, retinoblastoma family members, differentiation regulated transcription factor polypetide-1, and cyclin H. These cell-cycle regulators were not eliminated when the peptide inhibitor was used. Dysfunction of cell-cycle regulators may play a critical role in the signal transduction pathway for activation of FTY720 mediated apoptosis. PMID- 12723593 TI - (+)-Menthol and its hydroxy derivatives, novel fungal monoterpenols from the fusicoccin-producing fungi, Phomopsis amygdali F6a and Niigata 2. AB - In our search for new fusicoccins of unique diterpene glucosides from Phomopsis amygdali, we found that a fragrant substance was formed in the early stage of fusicoccin fermentation. This fragrant constituent was isolated and identified as (+)-menthol, which is a novel fungal metabolite as the enantiomer of well-known peppermint (-)-menthol. (+)-7-Hydroxymenthol and new (+)-(6S)-hydroxymenthol were also isolated and identified as fungal metabolites. In addition, p-menthanetriol, which has been reported as the first fungal monoterpene from the fungus, was also isolated. The possible biosynthetic relationship of these metabolites is discussed. PMID- 12723594 TI - High-throughput PCR screening of genes for three-component regulatory system putatively involved in quorum sensing from low-G + C gram-positive bacteria. AB - Quorum sensing of gram-positive bacteria is often regulated by three-component regulatory system composed of autoinducing peptide, sensor kinase and response regulator. We used PCR to study a gene cassette encoding this three-component regulatory system. Degenerate primers were designed from consensus amino acid sequences in the HPK10 subfamily, mostly involved in quorum sensing. Products amplified from genomic DNA of Lactobacillus, Enterococcus, and Clostridium species were cloned and sequenced; their deduced amino acid sequences were similar to those of members of the HPK10 subfamily. Complete genes for the putative gene cassette were cloned by inverse PCR from L. paracasei E93490 and L. plantarum WCFS6. Phylogenetic analysis grouped the cloned putative HPKs into the HPK10 subfamily. These results indicated the usefulness of this high-throughput gene screening and suggested that the three-component regulatory gene cassette are widely present. PMID- 12723595 TI - Characterization of aspartate aminotransferase from the cyanobacterium Phormidium lapideum. AB - Aspartate aminotransferase (AspAT) was purified to homogeneity from cell extracts of the non-N2-fixing cyanobacterium Phormidium lapideum. The NH2-terminal sequence of 25 amino acid residues was different from the sequences of the subfamily Ialpha of AspATs from eukaryotes and Escherichia coli, but it was similar to sequences of the subfamily Igamma of AspATs from archaebacteria and eubacteria. The enzyme was most active at 80 degrees C and was stable at up to 75 degrees C. Thermal inactivation (60-85 degrees C) of the enzyme followed first order kinetics, with 2-oxoglutarate causing a shift of the thermal inactivation curves to higher temperatures. However, at 25 degrees C the kcat of P. lapideum AspAT was nearly equal to the values of AspATs from mesophilic organisms. The enzyme used L-aspartate and L-cysteine sulfinate as amino donors and 2 oxoglutarate as an amino acceptor. The Km values were 5.0 mM for L-aspartate, 5.7 mM for L-glutamate, 0.2 mM for 2-oxoglutarate, and 0.032 mM for oxaloacetate. PMID- 12723596 TI - Regioselective glucosylation of pyridoxine by microorganisms. AB - Microorganisms from culture collections and isolates from nature were screened for the ability to catalyze the regioselective glucosylation of pyridoxine (PN) to produce pyridoxine 5'-alpha-D-glucoside (PN-5'-alpha-G) or pyridoxine 4'-alpha D-glucoside (PN-4'-alpha-G). Transglucosylation activity specific to 5'-position of PN was found in fungi belonging to genera such as Coriolus and Verticillium, and activity at the 4'-position of PN was found in bacteria belonging to genera such as Bacillus and Serratia. From 100 mM PN, intact cells of Verticillium dahliae TPU 4900 produced 42 mM (13.9 mg/mL) PN-5'-alpha-G after 70 h of reaction. Intact cells of Bacillus cereus TPU 5504 produced 33 mM (10.9 mg/mL) PN 4'-alpha-G after 19 h of reaction. The selectivities for 5'- and 4'-positions were 80% and 90%, respectively. PMID- 12723597 TI - Improvement in 5'-position-selective glucosylation of pyridoxine by Verticillium dahliae TPU 4900. AB - Optimization of culture and reaction conditions for 5'-position-selective transglucosylation to pyridoxine by Verticillium dahliae TPU 4900 was investigated. V. dahliae TPU 4900 had high transglucosylation activity when grown with soluble starch as a carbon source and organic nitrogens such as Esusan meat as a nitrogen source at 15-20 degrees C. Both the yield of pyridoxine 5'-alpha-D glucoside (PN-5'-alpha-G) and the 5'-position-selectivity reached a maximum when an intact-cell reaction was done at 50-60 degrees C and pH 7 with additions of dextrin. The transglucosylation activity in culture broth was 71 times with the optimization of culture conditions that under the conditions used for screening. The productivity of PN-5'-alpha-G synthesis was 6.9 times that under the initial conditions when the reaction conditions of intact cells were optimized. From 1000 mM (206 g/L) pyridoxine hydrochloride, PN-5'-alpha-G was synthesized to the concentration of 300 mM (98.4 g/L as PN-5'-alpha-G) with 5'-selectivity of 85% in 53 h by intact cells of V. dahliae TPU 4900. PMID- 12723598 TI - Characterization of a Saccharomyces cerevisiae mutant with pseudohyphae and cloning of a gene complementing the mutation. AB - Screening for morphological mutants of a haploid strain of Saccharomyces cerevisiae was done on the basis of their cell-shape on a solid medium containing isoamyl alcohol, which causes cell elongation, to obtain information on the morphogenesis. Mutant J19, which had pseudohyphae in liquid medium even in the absence of isoamyl alcohol, had many elongated cells. Few reports exist of haploid cells growing as pseudohyphae in liquid culture. Cell-wall analysis showed that J19 had ordinary amounts of alkali-insoluble glucan and chitin, but that isoamyl alcohol in the medium caused structural changes in the cell wall. Addition of a DNA fragment that included the wild-type SCL1 gene to J19 complemented its morphological phenotype. Sequencing of J19 SCL1 showed that the glycine at position 226 in the Scl1 protein had been replaced by asparatic acid, suggesting that this mutation in the protein, a subunit of proteasomes, may be involved in the morphological change. PMID- 12723600 TI - Acarviosine-simmondsin, a novel compound obtained from acarviosine-glucose and simmondsin by Thermus maltogenic amylase and its in vivo effect on food intake and hyperglycemia. AB - Simmondsin was modified with acarviosine-glucose using the transglycosylation activity of Thermus maltogenic amylase to synthesize a novel compound with both antiobesity and hypoglycemic activity. The LC/MS and 13C NMR analyses confirmed that the structure of the major transglycosylation product was acarviosine simmondsin (Acv-simmondsin), in which acarviosine was attached to the glucose moiety of simmondsin by an alpha-(1,6)-glycosidic linkage. It was found that Acv simmondsin was a potent competitive inhibitor of alpha-glucosidase with the Ki value of 0.69 microM and a mixed type inhibitor of alpha-amylase with the Ki and KI of 20.78 microM and 26.31 microM, respectively. The administration of Acv simmondsin (0.1 g/100 g diet/day) to mice for 5 days significantly reduced food intake by 35%, compared to 25% with simmondsin in control obese mice. Acv simmondsin (50 mg/kg BW) suppressed the postprandial blood glucose response to sucrose (1 g/kg BW) by 74%, compared to 71% with acarbose, in normal rats. PMID- 12723599 TI - Maltosyl-erythritol, a major transglycosylation product of erythritol by Bacillus stearothermophilus maltogenic amylase. AB - This study was done to modify erythritol to change its physicochemical and sensory properties. Erythritol, a four-carbon sugar alcohol, was transglycosylated by Bacillus stearothermophilus maltogenic amylase with maltotriose as a donor molecule. The presence of various transglycosylation products of erythritol was confirmed by TLC and high performance ion exchange chromatography (HPIC). The major transfer product was purified by gel filtration chromatography on Bio-Gel P-2. Examination by LC-MS, matrix-assisted laser desorption ionisation-time of flight mass spectrometry (MALDI-TOF-MS), and 13C NMR showed that the major transfer product was maltosyl-erythritol. Results of 13C NMR of maltosyl-erythritol suggested that linkage was formed between the C1 carbon of glucose unit in maltose and either one of the two carbon atoms of the terminal hydroxyl groups of erythritol, so that a mixture of 1-O- and 4-O-alpha maltosyl-erythritol was produced. The sweetness of maltosyl-erythritol was about 40% that of sucrose, and its negative sensory properties were less than those of erythritol. PMID- 12723601 TI - Histidine-114 at subsites E and F can explain the characteristic enzymatic activity of guinea hen egg-white lysozyme. AB - The courses of the reaction catalyzed by guinea hen egg-white lysozyme (GHL), in which Asn113 and Arg114 at subsites E and F in hen egg-white lysozyme (HEL) are replaced by Lys and His, respectively, was studied with the substrate N acetylglucosamine pentamer, (GlcNAc)5. Although GHL was found to retain the main chain folding similar to HEL as judged from CD spectroscopy, the courses of GHL showed increased production of (GlcNAc)4 and reduced production of (GlcNAc)2 when compared with HEL. To identify critical residue(s) involved in the alteration in the courses of GHL, two mutant enzymes as to subsites E and F in HEL, N113K and R114H, were prepared by site-directed mutagenesis. Kinetic analysis of these mutants revealed that the mutation of Asn113 to Lys had little effect on the courses of HEL, while the Arg114 to His mutation completely reproduced the courses of GHL, demonstrating that His114 in GHL is the key residue responsible for the characteristic courses of GHL. Computer simulation of the reaction courses of the R114H mutant revealed that this substitution decreased not only the binding free energies for subsites E and F, but also the rate constant of transglycosylation. The Arg residue at position 114 may play an important role in the transglycosylation activity of HEL. PMID- 12723602 TI - Characterization of the Prr1 response regulator with special reference to sexual development in Schizosaccharomyces pombe. AB - The histidine (His)-to-Aspartate (Asp) phosphorelay is a paradigm of intracellular signaling systems through protein phosphorylation in both prokaryotes and eukaryotes. The fission yeast Schizosaccharomyces pombe has three histidine kinases (Phk1/Mak2, Phk2/Mak3, and Phk3/Mak1), together with two response regulators (Mcs4 and Prr1). The results of recent extensive studies suggested that these His-to-Asp phosphorelay components are involved in oxidative stress responses through the transcriptional regulation of several scavenger genes for toxic free radicals. It was also suggested that they were somehow implicated in control of both the mitotic and meiotic cell proliferations. Among these S. pombe His-to-Asp phosphorelay components, however, the function of Prr1 is less clear. We here characterized a mutant, named prr1-D418N, specifying an altered Prr1 protein that presumably acts as a gain-of-function (or constitutive active) mutant, with special reference to sexual development. The mutant cells showed a striking phenotype in that they underwent mating even in a nitrogen sufficient medium, under which conditions the wild-type cells hardly did so. Furthermore, the mutant cells underwent mating very rapidly in a nitrogen deficient medium. Under anaerobic (or micro-aerobic) growth conditions, the wild type cells were not capable of undergoing sexual development even in a nitrogen deficient medium. The prr1-D418N cells underwent mating efficiently under such anaerobic growth conditions. Taken these together, it was suggested that the function of Prr1 is closely linked to the well-characterized signaling pathways for induction of the sexual development, in a way that this response regulator regulates a critical step of the initiation of meiosis through activating the transcription of ste11+, mam2+, and mei2+, in S. pombe. PMID- 12723603 TI - Characterization of rice functional monosaccharide transporter, OsMST5. AB - cDNA of a monosaccharide transporter in rice, OsMST5 (Oryza sativa monosaccharide transporter 5) was cloned and its sugar transport activity was characterized by heterologous expression analysis. The amino acid sequence and topology were similar to the sequences and topology of other plant monosaccharide transporters. Yeast cells co-expressed with OsMST5 cDNA transported some monosaccharide substrates. The transport rate increased when ethanol as an electron donor was added, so the transporter was an energy-dependent active one. Most of the OsMST5 was expressed in panicles before pollination, indicating that it is associated with pollen development in rice. PMID- 12723604 TI - The N-terminal sequence of paratropomyosin binding fragments from beta-connectin. AB - In order to clarify the position where paratropomyosin binds to connectin in the A-I junction region of a sarcomere, chicken beta-connectin was digested by Staphylococcus aureus V8 protease under denaturing conditions and the digested peptides were electrophoretically separated. Five peptides, 150-kDa, 100-kDa, 70 kDa, and 43-kDa fragments, were simultaneously detected by biotinylated paratropomyosin and an anti-connectin monoclonal antibody. The N-terminal sequence of the 43-kDa fragment was found to be YQFRVYAVNK, similar to the sequence of 7556-7565 amino acids in the I51 fibronectin type 3 domain that was located at the A-I junction region of human cardiac titin/connectin. Therefore, we propose that paratropomyosin binds to the 43-kDa fragment from beta-connectin at the A-I junction region in both living muscle and in muscle immediately postmortem, and the N-terminus of the 43-kDa fragment is localized in the I51 domain. PMID- 12723605 TI - Enzymatic properties of glutamine 32 mutants of RNase Rh from Rhizopus niveus, a trial to alter the most preferential inter-nucleotidic linkages of RNase Rh. AB - In order to investigate the effects of mutation of Gln32, a component of a base recognition site (B2 site) of a base-nonspecific RNase from Rhizopus niveus, we prepared several enzymes mutant at this position, Q32F, Q32L, Q32V, Q32T, Q32D, Q32N, and Q32E, and their enymatic activities toward RNA and 16 dinucleoside phosphates were measured. Enzymatic activities of the mutant enzymes towards RNA were between 10-125% of the native enzyme. From the rates of hydrolysis of 16 dinucleoside phosphates by mutant enzymes, we estimated the base specificity of both B1 and B2 sites. The results indicated that mutation of Gln32 to Asp, Asn, and Glu caused the B2 site to prefer cytosine more and to a less extent, to prefer uracil (Q32N), and that Q32F made the enzyme more guanine-base preferential. The results suggested that we are able to construct an enzyme that preferentially cleaves internucleotidic linkages, at the 5'-side of cytosine residues (Q32D, Q32N, and Q32E) and guanine residues (Q32F and Q32T), thus, cleaves purine-C(Q32D, Q32N, Q32E) and GpG and ApG (Q32F, and Q32T) most easily. The results seemed to suggest converting a base-non-specific RNase to a base specific one. PMID- 12723606 TI - Effects of bovine alpha-lactalbumin on gastric defense mechanisms in naive rats. AB - We recently investigated the effects of the major proteins in cow's milk on gastric mucosal injuries in rat ulcer models. We found that alpha-lactalbumin (alpha-LA) has marked preventive effects against gastric mucosal injuries and that prostaglandin (PG) synthesis may contribute to these effects [Matsumoto et al., Biosci. Biotechnol. Biochem., 65, 1104-1111, 2001]. In this study, we investigated the effects of alpha-LA on several defense mechanisms of gastric mucosa by evaluating gastric PGE2 content, gastric mucin content, gastric luminal pH, gastric fluid volume, and gastric emptying in naive rats. Oral administration of alpha-LA (200, 500, and 1000 mg/kg) elevated endogenous PGE2 levels in gastric tissue and increased the gastric mucin contents of both the gastric fluid and the adherent mucus gel layer. In addition to these PG-related responses, alpha-LA also caused PG-independent responses such as elevation of gastric luminal pH, increase in gastric fluid volume, and delay in gastric emptying. These responses were observed to be dose-dependent (200-1000 mg/kg of alpha-LA). Thus, we demonstrated that alpha-LA enhances both PG-dependent and PG-independent gastric defense mechanisms in naive rats. Both of these mechanisms are probably involved in its gastroprotective action. PMID- 12723607 TI - Cloning of the xylitol dehydrogenase gene from Gluconobacter oxydans and improved production of xylitol from D-arabitol. AB - Xylitol dehydrogenase (XDH) was purified from the cytoplasmic fraction of Gluconobacter oxydans ATCC 621. The purified enzyme reduced D-xylulose to xylitol in the presence of NADH with an optimum pH of around 5.0. Based on the determined NH2-terminal amino acid sequence, the gene encoding xdh was cloned, and its identity was confirmed by expression in Escherichia coli. The xdh gene encodes a polypeptide composed of 262 amino acid residues, with an estimated molecular mass of 27.8 kDa. The deduced amino acid sequence suggested that the enzyme belongs to the short-chain dehydrogenase/reductase family. Expression plasmids for the xdh gene were constructed and used to produce recombinant strains of G. oxydans that had up to 11-fold greater XDH activity than the wild-type strain. When used in the production of xylitol from D-arabitol under controlled aeration and pH conditions, the strain harboring the xdh expression plasmids produced 57 g/l xylitol from 225 g/l D-arabitol, whereas the control strain produced 27 g/l xylitol. These results demonstrated that increasing XDH activity in G. oxydans improved xylitol productivity. PMID- 12723608 TI - Functional analysis of individual oligosaccharide chains of Sendai virus hemagglutinin-neuraminidase protein. AB - The roles of N-linked glycosylation in the intracellular transport and biological activities of the Sendai virus hemagglutinin-neuraminidase (HN) protein were studied. The protein contains four potential N-glycosylation sites: N77, N448, N499, and N511. By site-directed mutagenesis of these positions, the mature protein contained three N-linked oligosaccharides attached to N77, N499, and N511. The role of each added oligosaccharide in the structure and functions of the protein was identified by characterization of surface expression, hemadsorption, and neuraminidase activities of the corresponding mutant proteins. Elimination of the sites of N499 and N511 had the most detrimental effect, decreasing surface expression and hemadsorption. Elimination of the sites of N77 and N448 had similar but weaker effects. Mutants missing the sites of N499 and N511 were not able to induce syncytia formation in cells expressing mutant HN proteins and the fusion protein. Therefore, the N-linked oligosaccharides attached to N499 and N511 were important for intracellular transport and for the fusion promotion. PMID- 12723609 TI - Isotope ratio analysis by HRGC-MS of monoterpene hydrocarbons from citrus essential oils. AB - The isotope ratio of monoterpene hydrocarbons in citrus essential oils of different origins was measured by ordinary high-resolution gas chromatography mass spectrometry (HRGC-MS). The isotope ratio (Ir) was determined by the ratio of the isotope peak intensity (m/z 137) to the molecular mass peak intensity (m/z 136) of the monoterpene hydrocarbons. The accuracy of Ir was examined by measuring monoterpene hydrocarbon standards and 13C-labeled compounds. The isotope fingerprints based on the values of monoterpene hydrocarbons from lemon, lime and yuzu essential oils were determined. These citrus essential oils were also discriminated by a principal component analysis of their Ir data. The characteristic vectors showed that alpha-terpinene, beta-pinene and beta phellandrene were important components for distinguishing between the citrus species. It is suggested that this technique will be applicable to evaluate the quality, genuineness and origin of citrus fruits and their products. PMID- 12723610 TI - Functional analysis of a beta-ketoacyl-CoA synthase gene, MpFAE2, by gene silencing in the liverwort Marchantia polymorpha L. AB - We have isolated a beta-ketoacyl CoA synthase (KCS) gene, MpFAE2, from a liverwort, Marchantia polymorpha, and identified its substrate specificity using the technique of dsRNA-mediated gene silencing and overexpression. KCS catalyzes an essential reaction in the fatty acid elongation process, i.e., condensation of malonyl-CoA with acyl-CoA. By introducing a construct with a hairpin structure containing a partial MpFAE2 gene, the level of the MpFAE2 gene expression was suppressed constitutively. The transgenic plants showed a specific accumulation of fatty acid 18:0. In contrast, in transgenic M. polymorpha plants overexpressing the MpFAE2 gene, fatty acid 22:0 is accumulated. These results indicate that the MpFAE2 gene product catalyzes the elongation steps of 18:0 to 20:0 and possibly also of 20:0 to 22:0. PMID- 12723611 TI - Cloning of structural gene of Deinococcus radiodurans UV-endonuclease beta. AB - To characterize its enzymic property we cloned and sequenced the gene of Deinococcus radiodurans encoding UV-endonuclease beta, an alternative enzyme to UvrABC repairing damaged DNA. Amino acid substitutions were found in UV-sensitive mutants. The putative amino acid sequence had some similarity with those of eukaryotic UV-endonucleases and with a sequence found in a protein data base of Bacillus subtilis. PMID- 12723612 TI - Effects of adding some dietary fibers to a cystine diet on the activities of liver antioxidant enzymes and serum enzymes in rats. AB - This study investigates whether some dietary fibers can the toxicity due to cystine added to the diet. Wistar rats were investigated for the effects of adding pectin, sugar beet fiber or konjac mannan to a cystine diet on the growth rate and on the activities of liver antioxidant enzymes and serum enzymes. The addition of pectin, sugar beet fiber or konjac mannan to the cystine diet resulted in a significant increase in both the food intake and body weight gain. Feeding the cystine diet caused lower activities of total and Cu,Zn-superoxide dismutase, and of catalase in the liver. The addition of pectin to the cystine diet counteracted the activities of the total and Cu,Zn-superoxide dismutase, and of catalase in liver. Of the dietary fibers tested, konjac mannan prevented the elevation of the two enzyme activities in the serum induced by feeding the cystine diet, indicating that this fiber might have the ability to alleviate hepatic damage due to dietary cystine. PMID- 12723614 TI - Purification and characterization of extracellular beta-galactosidase secreted by supension cultured rice (Oryza sativa L.) cells. AB - A beta-galactosidase was purified 1300-fold by lactosyl-Sepharose 4B and Sephacryl S-200 column chromatographies from the cultured medium of a rice-cell suspension. The purified enzyme appeared as 47 kD and 40 kD polypeptides on SDS PAGE and had a specific activity of 65.1 units/mg. Optimum activity was observed at pH 3.5 and 60 degrees C. The enzyme released galactose from galactoxyloglucan and pectic galactans. PMID- 12723613 TI - Genomic organization of the S-locus region of Brassica. AB - To gain some insights into the structure of the S-locus and the mechanisms that have kept its diversity, a 75-kb genomic fragment containing the self incompatibility (S) locus region was isolated from the S12-haplotype of Brassica rapa and compared with those of other S-haplotypes. The region around the S determinant genes was highly polymorphic and filled with S-haplotype-specific intergenic sequences. The diverse genomic structure must contribute to the suppression of recombination at the S-locus. PMID- 12723615 TI - Mushroom tyrosinase inhibitory activity of esculetin isolated from seeds of Euphorbia lathyris L. AB - A tyrosinase inhibitor was isolated from the seeds of Euphorbia lathyris L. by bioassay-guided fractionation and purification, using silica gel column chromatography. It was identified as esculetin by comparing its physical properties and spectral data with those of an authentic sample. The IC50 value of esculetin in the mushroom tyrosinase activity test was 43 microM. The kinetic study indicates that esculetin exhibited competitive inhibition against the oxidation of 3-(3,4-dihydroxyphenyl)-alanine by mushroom tyrosinase. The structure-activity relationships among five esculetin analogs suggest that hydroxyl groups at the C6 and C7 positions of the coumarin skeleton played an important role in the expression of tyrosinase inhibitory activity. PMID- 12723617 TI - Continuous cell-free protein synthesis directed by messenger DNA and catalyzed by extract of Thermus thermophilus HB27. AB - Polypeptide synthesis directed by DNA as the messenger in a cell-free system of Thermus thermophilus was investigated. Polypeptides were synthesized with the addition of neomycin in the presence of DNA catalyzed by the cell extract. The stability of messenger DNA was greater than that of messenger RNA. Continuous cell-free translation with messenger DNA produced polypeptides at the rate of more than 8 microg/h in the presence of spermine. PMID- 12723616 TI - Inhibitory activity of analogs of AM-toxin, a host-specific phytotoxin from the Alternaria alternata apple pathotype, on photosynthetic O2 evolution in apple leaves. AB - The effect of the host-specific phytotoxins, AM-toxins, on the photosynthetic activity of leaves from susceptible apple cultivars was investigated by using an oxygen electrode. The photosynthetic O2 evolution was inhibited by AM-toxin I in a host-specific manner. The inhibitory activity of several AM-toxin analogs against photosynthesis was also evaluated and the findings were correlated with their necrosis-inducing activity. PMID- 12723618 TI - Formation of 3'-O-beta-galactosyl compounds of 5-bromouridine by Sporobolomyces singularis. AB - Two new compounds of 5-bromouridine, 3'-O-(beta-D-galactopyranosyl)-5 bromouridine and 3'-O-[beta-D-galactopyranosyl-(1-->4)-beta-D-galactopyranosyl]-5 bromouridine, were found to be selectively formed in a high yield in a culture filtrate of Sporobolomyces singularis, when grown on a medium containing lactose and 5-bromouridine. PMID- 12723619 TI - Cloning and overexpression of beta-N-acetylglucosaminidase encoding gene nagA from Aspergillus oryzae and enzyme-catalyzed synthesis of human milk oligosaccharide. AB - We isolated a beta-N-acetylglucosaminidase encoding gene from the filamentous fungus Aspergillus oryzae, and designated it nagA. The nagA gene encoded a polypeptide of 600 amino acids with significant similarity to glucosaminidases and hexosaminidases of various eukaryotes. A. oryzae strain carrying the nagA gene under the control of the improved glaA promoter produced large amounts of beta-N-acetylglucosaminidase in a wheat bran solid culture. The beta-N acetylglucosaminidase was purified from crude extracts of the solid culture by column chromatographies on Q-Sepharose and Sephacryl S-200. This enzyme was used for synthesis of lacto-N-triose II, which is contained in human milk. By reverse hydrolysis reaction, lacto-N-triose II and its positional isomer were synthesized from lactose and D-N-acetylglucosamine in 0.21% and 0.15% yield, respectively. PMID- 12723620 TI - Targeted gene disruption of the neuronal calcium sensor 1 homologue in rice blast fungus, Magnaporthe grisea. AB - We isolated a neuronal calcium sensor 1/frequenin-like gene, Mg-NCS-1, from Magnaporthe grisea and evaluated the phenotypes of null-mutants of the gene. The putative Mg-NCS-1 protein showed high similarity to the other NCS-1 proteins. The null-mutants had normal growth and pathogenicity similar to the parental strain, but their growth was suppressed in high concentrations of Ca2+ or acidic conditions. PMID- 12723621 TI - Molecular cloning of rat transcription factor YY1. AB - YY1 is a ubiquitously expressed multifunctional transcription factor that is involved in both positive and negative regulation of gene expression as well as initiation of transcription. Here, we isolated cDNA encoding a full-length open reading frame (ORF) of rat YY1. Rat YY1 is composed of 411 amino acid residues and its amino acid sequence is 97.6% identical to that of mouse YY1 and 97.8% identical to that of human YY1. The transactivating abilities of wild-type rat YY1 and four truncated mutant forms of YY1 were examined by transient reporter assays. When residues 114-193, which sequence includes a portion of the activation region and most of the Gly/Lys-rich region, were lacking, transactivation activity decreased somewhat, but the further deletion in the activation region (of residues 56-113) did not cause further decrease of the activity. On the other hand, N-terminus of the activation region (1-78/100-106) did not have transactivation activity by itself as well as synergistic activity with an erythroid specific transcription factor GATA-1. PMID- 12723622 TI - Total synthesis of 0231B, an inhibitor of 3alpha-hydroxysteroid dehydrogenase produced by Streptomyces sp. HKI0231. AB - The new inhibitors of 3alpha-hydroxysteroid dehydrogenase, 0231A 1 and 0231B 2, have a unique benz[c,d]indol-3(1H)-one structure in their molecules. In our advanced studies on indole chemistry, we have developed an efficient synthetic method for benz[c,d]indol-3(1H)-one derivatives. We report here its application to the synthesis of 0231B in 10 steps (8.1% overall yield) from 6-methylindole 8 by introducing an acyl group into the 3-position of the indole nucleus, cyclization of the side chain at the 3-position to the 4-position and subsequent elimination of the phenyl group, and conjugate addition of the substituted phenyl group. PMID- 12723623 TI - Gnetol as a potent tyrosinase inhibitor from genus Gnetum. AB - Gnetol (2,3',5',6-tetrahydroxy-trans-stilbene), a naturally occurring compound particularly found in the genus Gnetum, had a strong inhibitory effect on murine tyrosinase activity. Gnetol (IC50, 4.5 microM) was stronger than kojic acid (IC50, 139 microM) as a standard inhibitor for murine tyrosinase activity. Moreover, gnetol significantly suppressed, melanin biosynthesis in murine B16 melanoma cells. PMID- 12723624 TI - RCVS issues guidance on dealing with abuse in animals and humans. PMID- 12723625 TI - Practice standards: decisions for the future. PMID- 12723626 TI - Handling wildlife casualties. PMID- 12723627 TI - Experiences of phocine morbillivirus. PMID- 12723628 TI - Randomised, double-blind, placebo-controlled parallel group study of P54FP for the treatment of dogs with osteoarthritis. AB - P54FP is an extract of Indian and Javanese turmeric, Curcuma domestica and Curcuma xanthorrhiza respectively, which contains a mixture of active ingredients including curcuminoids and essential oils. A randomised, double-blind, placebo controlled, parallel group clinical trial of P54FP as a treatment for osteoarthritis of the canine elbow or hip was conducted to assess its efficacy and safety. Sixty-one client-owned dogs with osteoarthritis were recruited through first-opinion practices and examined at a single centre. After a two-week wash-out period, they were randomly allocated to receive P54FP or a placebo orally twice daily for eight weeks, and were re-examined after four, six and eight weeks of treatment. The effectiveness of the treatment was assessed in terms of the peak vertical force (PVz) and vertical impulse of the affected limbs, as measured with a force platform, by clinical assessments of lameness and joint pain by the investigators, and overall assessments of the response to treatment by the investigators and the owners. The results from 25 P54FP-treated dogs and 29 placebo-treated dogs showed that there was no statistically significant difference between the groups in terms of the PVz of the affected limb. The investigators' overall assessment showed a statistically significant treatment effect in favour of P54FP (P=0.012), but the owners' assessment just failed to reach statistical significance (P=0.063). No serious adverse effects were recorded, but two P54FP-treated dogs and four placebo-treated dogs were withdrawn from the study because their condition deteriorated. PMID- 12723629 TI - Classical swine fever in wild boar in the Lombardy region of Italy from 1997 to 2002. AB - In 1997, outbreaks of classical swine fever occurred in wild boar and domestic pigs in the Italian province of Varese (Lombardy region). Statutory animal health measures were imposed, and a control plan aimed at preventing the virus from spreading outside the infected hunting zone of 108 km2 was implemented. However, in 1998, virologically positive wild boar were found outside this zone, including within Swiss territory. The infected zone was then enlarged to include the three other hunting areas of the northern province of Varese and the neighbouring Swiss territory. Italian and Swiss veterinary services initiated a joint policy to control the disease by hunting young wild boar and conserving older, immune animals. The percentage of virologically positive animals gradually decreased, the last being found in July 2000. The seroprevalence initially increased, but in 2000 and 2001 it decreased from 42.2 per cent to 8.8 per cent, indicating that the wild boar population had had no further contact with the virus. PMID- 12723630 TI - In vitro activity of 10 antimicrobial agents against bacteria isolated from cows with clinical mastitis. AB - The susceptibility of 495 strains of bacteria, recently isolated in France from cows with clinical mastitis, to 10 antimicrobial agents--penicillin G, cloxacillin, oxacillin, cephalexin, cefazolin, cephapirin, cefquinome, neomycin, ampicillin and colistin--was determined by measuring their minimum inhibitory concentrations (MICS). Overall, the levels of resistance were very low except for staphylococci and penicillin G. The 167 streptococcal strains were susceptible to all of the beta-lactams tested, but six (3-6 per cent) were highly resistant to neomycin. Of the 171 staphylococcal isolates, 36.2 per cent were resistant to penicillin G, one strain of Staphylococcus sciuri was classified as methicillin resistant, but they were all susceptible to neomycin. None of the 122 strains of Escherichia coli was resistant to colistin, but 12 had high MIC values for one or more of the cephalosporins. PMID- 12723631 TI - First PCR-confirmed report of a Neospora caninum-associated bovine abortion in Austria. PMID- 12723632 TI - Mycobacterium kansasii infection in a Chinese soft shell turtle (Pelodiscus sinensis). PMID- 12723633 TI - Extension of the VLA surveillance network. PMID- 12723634 TI - Daft lamb disease. PMID- 12723635 TI - Vaccine-associated feline sarcoma. PMID- 12723636 TI - Testing for equine arteritis virus. PMID- 12723637 TI - FMD control strategies. PMID- 12723638 TI - FMD control strategies. PMID- 12723639 TI - Bird tick survey. PMID- 12723641 TI - Probiotics: considerations for human health. AB - Evidence for the role of probiotics in maintenance of health or prevention of disease is mounting and is supported in some cases by blinded, placebo-controlled human trials. Today, in an era of antibiotic-resistant pathogens and other looming microbial threats, the value of prevention of infection is recognized. Probiotics may play an important role in helping the body protect itself from infection, especially along the colonized mucosal surfaces of the gastrointestinal tract. Probiotic products are available in many different forms worldwide, including pills, powders, foods, and infant formula. In some cases, general health claims are made that cannot be substantiated for the specific strains and levels being used and consumers must therefore beware. PMID- 12723640 TI - Triple trouble: the role of malnutrition in tuberculosis and human immunodeficiency virus co-infection. AB - Worldwide, the number of individuals who are co-infected with human immunodeficiency virus (HIV) and tuberculosis is increasing greatly. The "triple trouble" of HIV and tuberculosis infection and malnutrition may put those infected at greater risk than those with any of the three conditions alone. Further investigation is needed to evaluate the prophylactic and therapeutic potential of nutritional interventions for co-infection with HIV and tuberculosis. PMID- 12723642 TI - Ghrelin: update 2003. AB - Ghrelin is a recently described peptide hormone that is secreted by endocrine cells in the gastrointestinal tract. Although its initial discovery was as a novel growth hormone secretagogue, it has been found to regulate feeding behavior by modulating expression levels of orexigenic peptides in the hypothalamus. Ghrelin has been implicated in the coordination of energy balance and weight regulation, and its dysregulation may be important in obesity. Ghrelin also has several other physiologic actions besides potential regulation of food intake that are described in this brief review. PMID- 12723644 TI - Prevalence of vitamin D insufficiency in Canada and the United States: importance to health status and efficacy of current food fortification and dietary supplement use. AB - Several recent studies have identified a surprisingly high prevalence of vitamin D insufficiency in otherwise healthy adults living in Canada and the United States. Most striking are the effects of latitude, season, and race. Also noteworthy is that dietary vitamin D is not reaching the population in greatest need, nor is it very protective against insufficiency. Fluid milk, as the predominant vehicle for vitamin D fortification, is apparently not very effective in staving off vitamin D insufficiency in adults in all populations at all times of the year. PMID- 12723643 TI - Body mass index and mortality in asian populations: implications for obesity cut points. AB - Investigators have questioned whether body mass index (BMI, kg/m2) cut-points for obesity used in the United States and Europe are appropriate for Asian countries. A recent study examined the association between BMI and mortality in a population based cohort of Japanese men and women. These and other results did not indicate a need for lower cut-points in Asians. PMID- 12723645 TI - Severe acute respiratory syndrome-global alert. PMID- 12723646 TI - Molecular diagnosis of cystic fibrosis in Indian patients--a preliminary report. AB - AIM: The aim of the study was to screen for the common deltaF508 mutation and the poly T polymorphism and to determine their frequency in the cystic fibrosis transmembrane conductance regulator (CFTR) gene among the suspected CF cases referred to our clinical care centre for sweat chloride tests. METHODOLOGY: Sweat and EDTA blood samples were obtained from 23 clinically suspected cystic fibrosis (CF) cases. Sweat was estimated by pilocarpine iontophoresis procedure. Poly T polymorphism was detected by the multiplex-PCR based on ARMSTM technique and deltaF508 mutation by PCR-mediated site-directed mutagenesis method. RESULTS: Five cases, mainly with respiratory abnormalities and followed by steatorrhea had elevated sweat chloride levels (> 60 mmol/l), three of them, each with nutritional, respiratory and pancreatic abnormalities were borderline (40-60 mmol/l) and the remaining 15 clinically suspected CF cases had normal sweat chloride levels (< 40 mmol/l). The 9T variant was frequently observed (75%) in cases with elevated sweat chloride, including those exhibiting borderline values; with no 5T variant. The 7T was the most common variant (77%) observed in the cases with normal sweat chloride, with only one 5T variant (33%). Of the five cases with high sweat chloride, four cases were homozygous for deltaF508, whereas one was heterozygous with borderline sweat chloride, thus showing an overall frequency of 56.25% in the CF chromosome. DeltaF508 was found to be present with the 9T variant in all the instances. CONCLUSION: The presence of the 9T variant along with elevated sweat chloride levels can be used to predict a high risk of the individual harboring the severe deltaF508 mutation. It would be advisable to test for to the deltaF508 mutation along with the sweat chloride estimation in all the critically suspected CF cases diagnose CF with a higher degree certainty. PMID- 12723647 TI - A comparative evaluation of C-reactive protein as a short-term prognostic marker in severe unstable angina--a preliminary study. AB - OBJECTIVE: The present study was done to evaluate the short-term prognostic significance of C-reactive protein (CRP) in unstable angina (UA) and to compare it with other known prognostic markers of UA, as there is a paucity of data from our country. METHODOLOGY: This prospective study comprised of 44 UA patients (Braunwald Class II A, B) with age < 65 years, along with 40 age and sex matched healthy controls. Patients with h/o myocardial infarction (MI) in the preceding 1 month and evidence of infection, inflammation or neoplasm were excluded from the study. Complete clinical evaluation was done and presence of any prognostic variables of UA was noted (including Brauriwald high-risk variables). Apart from biochemical investigations and ECG, CRP was measured at the time of admission in the study group and controls using Microwell ELISA assay. Standard treatment protocol was followed for all patients and they were closely monitored during hospital stay and subsequently for 4 weeks for occurrence of any adverse cardiac events. RESULTS: There was a statistically significant difference in the mean level of CRP between study group (6.12 +/- 6.134 mg/L) and controls (1.52 +/- 0.751 mg/L); p = 0.000. Among the 44 patients, 19 (43%) experienced any one or more outcome measures. Of the three variables (i.e. ongoing chest pain, ST depression > or = 1 mm and CRP > or = 4 mg/L) which showed statistical significance on univariate analysis with respect to adverse outcome measures (p = 0.001, 0.027 and 0.000 respectively), only CRP > or = 4 mg/L and ST depression > or = 1 mm showed independent prognostic significance on multiple logistic regression analysis (p = 0.000 and 0.002 respectively). The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and relative risk (RR) for CRP > or = 4 mg/L were 78.9%, 96%, 93.75%, 85.74% and 6.56 (95% CI; 2.62 - 16.4) and that for ST depression > or = 1 mm were 89.47%, 64%, 65.38%, 88.89% and 5.88% (95% CI: 1.54 - 22.38), respectively. Trop T could be done in eight patients only. CONCLUSION: CRP > or = 4 mg/L by ELISA is an independent predictor of adverse cardiac outcome in severe UA in the short-term follow up and, hence, is useful for risk stratification of these patients. CRP has a higher specificity, PPV and overall RR for prediction of an outcome than ST segment depression, although it is less sensitive. PMID- 12723648 TI - Endothelium-dependent brachial artery flow mediated vasodilatation in patients with diabetes mellitus with and without coronary artery disease. AB - AIM OF THE STUDY: Endothelial function as assessed by brachial artery flow mediated vasodilatation (FMD) has been shown to be impaired in patients with coronary artery disease (CAD). Since diabetes mellitus (DM) has been considered to be CAD risk-equivalent, we sought to determine whether endothelial dysfunction is present in patients with DM independent of presence of CAD. METHODOLOGY: One hundred and ninety eight individuals were included in the study and divided into four groups: Group 1--patients with risk factors for CAD, but no DM or CAD; Group 2--patients with DM but no CAD; Group 3--patients with CAD but no DM and Group 4- patients with both DM and CAD. Brachial artery FMD assessment was performed once in all subjects and FMD was calculated as percentage increase in brachial artery diameter in response to increase in brachial artery flow. RESULTS: Mean FMD was significantly higher in Group 1 (7.03 +/- 2.87%) compared to the other three groups. Mean FMD in Group 2 (5.51 +/- 2.12%) was similar to that in Group 3 (4.56 +/- 2.70%; p value 0.195) but significantly higher than that in Group 4 (4.26 +/- 1.93%; p value 0.038). There was no statistically significant difference in mean FMD in Group 3 and Group 4 (p value 0.65). CONCLUSION: Endothelial function as assessed by FMD is significantly impaired in diabetics compared to non-diabetics in absence of CAD. In addition, similar degree of impairment in endothelial function is seen in diabetics without CAD and non-diabetic patients having CAD, implying CAD risk-equivalence of diabetes. PMID- 12723649 TI - Prevalence of anticardiolipin antibodies in various thrombotic conditions: a hospital-based study. AB - Thrombosis is one of the leading cause of death, globally, anticardiolipin antibodies (aCL) has been implicated as one of the most common acquired protein defect causing thrombosis. This study was undertaken to evince the incidence of aCL in various thrombotic settings. MATERIAL AND METHODS: Three hundred and two patients were retrospectively screened for the incidence of aCL The sera were screened for aCL IgG using enzyme linked immunosorbent assay. Detail clinical and epidemiological data were obtained from hospital records and clinical examination. RESULTS: Among the 302 patients, 134 (44.37%) were below the age of 40 years (juvenile-onset thrombosis), mean age being 35.3 years. High titres of aCL IgG was seen in 65 (20.77%) patients. Deep vein thrombosis (DVT), seen in 90 (29.8%) was the most common thrombotic condition. Other sites of thrombosis were coronary artery (19.2%), central nervous system territory arteries (17.21%) and peripheral arteries (5.29%); the incidence of aCL IgG in these sites were 13.79%, 25.0% and 18.75% respectively. CONCLUSION: aCL is the most common acquired thrombophilic defect. Epidemiological data of our population is required for evaluating the strategy for further research of thrombosis in this condition. PMID- 12723650 TI - Quality of life and diabetes integration among subjects with type 2 diabetes. AB - Diabetes integration indicates that a person with diabetes makes an appropriate emotional adjustment to the requirements of diabetic way of life. Diabetes integration and a sense of well-being are expected to be correlated. The aim of diabetes treatment should be to enable a patient to adjust himself to his being a diabetic person. This should ensure a sense of well-being. Aims of this study was to assess the role of diabetes integration and psychological factors in patients with type 2 diabetes. METHOD: A sample of 227 type 2 diabetes patients participated in the study. They were all in the upper middle class social stratum. Diabetes integration scale yields one composite score of adjustment, has 19 items applicable to both type 1 and type 2 diabetic subjects. The psychological well-being scale has 22 items, measures depression (6 items), anxiety (6 items), energy (4 items), positive well-being (6 items) and a general sense of well-being score by the whole test of 22 items. RESULTS: The psychological well-being subscales and the whole scale scores significantly correlated with diabetes integration scale (all P values were < 0.0001). However, we found that diabetes integration and the psychological well-being subscales were not significantly correlated with metabolic and other medical indices. There were gender differences in depression (p < 0.04), anxiety (p < 0.0001), energy (p = 0.004), positive well-being. (p = 0.02) and general sense of well-being (p < 0.0001), men fared in a better than women subjects. No such gender differences were found in diabetes integration score. CONCLUSION: Diabetic patients who integrate themselves and emotionally adjust to diabetes experience a psychological sense of well-being. PMID- 12723651 TI - Charcot foot--an update. PMID- 12723652 TI - The human genome project: its implications in clinical medicine. PMID- 12723653 TI - Plummer's nails. PMID- 12723654 TI - Heliotrope rash and 'V' sign in dermatomyositis. PMID- 12723655 TI - Sepsis and neurology. PMID- 12723656 TI - Idiopathic hypertrophic cervical pachymeningitis: a case report with 5 year follow up. AB - Idiopathic hypertrophic pachymeningitis is an extremely rare entity. It usually affects cranial meninges. The spinal form is further uncommon and presents as a chronic progressive disease. We describe a 42 year old female with isolated idiopathic hypertrophic cervical pachymeningitis who had a relapsing remitting course under observation for five years. Laminectomy and immunosuppressive therapy produced temporary and partial relief. The long term course and relevant literature is reviewed. PMID- 12723657 TI - Reactive arthritis, psoriasiform lesions and protein loosing enteropathy secondary to Strongyloidiasis. AB - Reactive arthritis is associated with demonstrable infection at a distant site without evidence of sepsis at the affected joint(s). We present a rare case reactive arthritis where no bacterial or chlamydial infections could be established, rather larvae of Strongyloides stercoralis could be demonstrated in the stool and duodenal biopsy. Reactive arthritis, psoriasiform lesions and malabsorption with hypoproteinaemia, responded to successful treatment with antihelminthic drugs. Early recognition and adequate treatment for gastrointestinal infections and infestations before complications is important. PMID- 12723658 TI - Pulmonary arteriovenous malformation treated with embolotherapy. AB - Pulmonary arteriovenous malformations (PAVM) represent pulmonary vascular anomalies and in majority of cases are congenital in origin. We report a case that presented with predominant complaints of dyspnoea on exertion and cyanosis. Clinical examination revealed a bruit in the left lower interscapular and infrascapular areas which led us to suspect PAVM. The diagnosis was established on the basis of computed tomography (CT) thorax with three-dimensional virtual reconstruction technology (3D VRT) images and pulmonary angiography. Patient was successfully treated by embolotherapy using steel coils, which resulted in 90% exclusion of the aneurysm from the rest of the systemic circulation. PMID- 12723660 TI - Mediastinitis presenting as pyrexia of unknown origin. AB - A 55 year old female was admitted as a case of pyrexia of unknown origin (PUO) of 2 months duration. She had developed throat ache, progressive dysphagia for both solids and liquids, dry cough and retrosternal pain for one week. Examination revealed fever, tachycardia, tachypnoea and a soft tissue warm tender, erythematous, non-fluctuant swelling in lower anterior neck with chest findings suggestive of bilateral pleural effusion. Plain X-rays of the neck and chest strengthened the clinical suspicion of cellulitis of lower neck with bilateral pleural effusion. CT scan confirmed the radiologic findings and also revealed pericardial effusion and thickening; small mediastinal lymphadenopathy and mediastinitis. Patient responded to parenteral antibiotics (ceftriaxone and metronidazole) and hydrocortisone with complete resolution in 10 days. PMID- 12723659 TI - Langerhans cell histiocytosis presenting as adult onset epilepsy. AB - A 37 year old man presented with recurrent secondarily generalized seizures from right partial onset since December 1999, MRI scan of brain (contrast study) revealed multiple enhancing lesions predominantly involving frontal, parietal and temporal regions. Left frontotemporal lesion was biopsied and histopathology confirmed it to be rare case of adult Langerhans cell histiocytosis. He was given short course of oral corticosteroids. Two years postoperative course on antiepileptic therapy alone is uneventful and repeat MRI brain and MR spectroscopy showed significant resolution of lesions. PMID- 12723661 TI - Aggressive behaviour of solid arrangement pattern in differentiated papillary carcinoma of thyroid. AB - The prognosis and overall outcome of solid growth pattern in differentiated papillary carcinoma of thyroid is controversial. While general consensus suggest outcomes similar to typical papillary carcinomas others demonstrate a more aggressive biology. We present a case of differentiated papillary carcinoma of thyroid with solid growth pattern presenting with neck swelling and widespread skeletal metastases. The areas of bone lesions showed avid 131I concentration on a postoperative large dose 131I whole body scan and was treated with 131I subsequently. The present case suggest that the solid architecture in a papillary thyroid carcinoma may be indicative of an aggressive clinical course, contrary to the common opinion that it does not adversely influence its biologic behaviour and thus emphasize the need to reexplore the prognostic significance of histopathologic subclassification along with an assessment of histologic grade and expression of molecular risk factors in this particular tumour subtype. PMID- 12723662 TI - Toxic thyroid adenoma and acromegaly: an unusual association. AB - Hyperthyroidism is seen in 3.5-26% of acromegalic subjects, and can occur through TSH-dependent or independent mechanisms. Thyrotoxicosis as the first presenting illness in acromegaly is particularly uncommon, as described in this patient who had both acromegaly and a toxic thyroid adenoma. PMID- 12723663 TI - Idiopathic hypereosinophilic syndrome manifesting as pulmonary oedema. AB - Idiopathic hypereosinophilic syndrome is a progressive and fatal disease if not treated effectively. We report this case since hypereosinophilia is an uncommon cause of pulmonary oedema. PMID- 12723664 TI - Duodenal diverticular haemorrhage in a patient taking NSAID. AB - We present a 55 years old male with severe anemia with history of pain abdomen for 2 weeks and malena of 10 days duration. He was taking NSAID (Diclofenac sodium). Upper GI endoscopy done twice did not reveal any abnormality. Upper gastrointestinal (UGI) barium series with small bowel follow through revealed a diverticulum on medial wall of second part of duodenum and there was evidence of ulcer in diverticulum. He underwent diverticulectomy. On follow up after 6 months patient was asymptomatic. PMID- 12723665 TI - Ineffectiveness of iron polymaltose in treatment of iron deficiency anemia. PMID- 12723666 TI - Myxoedema pellagra--a report of two cases. PMID- 12723667 TI - Dengue encephalitis. PMID- 12723668 TI - Failure of DOTS category--II treatment: a report on 3 cases. PMID- 12723669 TI - Trichinosis of psoas muscle. PMID- 12723670 TI - Observation of cases of hanging treated in medical wards. PMID- 12723671 TI - Methanol poisoning--a Chennai experience. PMID- 12723672 TI - Effectiveness versus efficacy: more than a debate over language. PMID- 12723674 TI - Factors associated with triathlon-related overuse injuries. AB - STUDY DESIGN: Descriptive correlational investigation. OBJECTIVES: To assess the incidence of, and potential risk factors associated with, overuse injury in triathlon. BACKGROUND: The sport of triathlon is rapidly increasing in popularity with a concomitant rise in the prevalence of injuries sustained by triathletes. METHODS AND MEASURES: The training and injury patterns of 131 triathletes were surveyed over a 10-week prospective period during the triathlon competition season. A complementary retrospective 6-month analysis of training history and prior overuse injuries was conducted. RESULTS: Fifty percent of triathletes sustained an injury in the 6-month preseason at an injury exposure rate of 2.5 per 1000 training hours. Thirty-seven percent were injured during the 10-week competition season at an injury exposure rate of 4.6 per 1000 training hours. Overuse accounted for 68% of preseason and 78% of competition season injuries reported. Increased years of triathlon experience, high running mileage, history of previous injury, and inadequate warming-up and cooling-down regimes appeared to have individual associations with injury incidence. When interactions were included in a multiple logistic regression model, increasing years of triathlon experience was the most significant predictor of preseason injury risk and a previous history of injury and high preseason running mileage increased the risk of injury during the competition season. CONCLUSIONS: The results indicate that in assessing triathletes, a full training and competition history is required by the sports clinician for a comprehensive assessment of the factors that may contribute to overuse injury. PMID- 12723673 TI - Deficits in detection of inversion and eversion movements among subjects with recurrent ankle sprains. AB - STUDY DESIGN: Observational cross-sectional design. OBJECTIVES: To determine whether a deficit exists in the ability to perceive inversion and eversion movements among subjects with recurrent ankle sprains. BACKGROUND: Although unproven, ankle sprains are thought to recur because proprioception is impaired subsequent to the original sprain. Proprioception has been widely studied, however, the specific property of movement perception has not been rigorously examined for both inversion and eversion movements. METHODS AND MEASURES: The ability to perceive passive inversion and eversion movements imposed at the relaxed ankle was measured in 39 subjects with recurrent ankle sprains (sprainers) and 30 subjects with no history of ankle injury (controls) by an assessor masked to subject group. The 70% detection level for movement perception was determined for 3 velocities, 0.1 degrees/s, 0.5 degrees/s, and 2.5 degrees/s, tested in random order. RESULTS: The 70% detection level for inversion and eversion movements improved with increasing velocity in both groups. At all velocities, the 70% detection level for movements imposed at the ankle of the sprainers was significantly worse than for controls. CONCLUSIONS: Perception of passive inversion and eversion movements imposed at the ankle was impaired in subjects with recurrent ankle sprains. The impairment was evident at the 3 velocities tested, 0.1 degrees/s, 0.5 degrees/s, and 2.5 degrees/s. PMID- 12723675 TI - Resident's case problem: diagnosis and treatment of posterolateral instability in a patient with lateral collateral ligament sprain. PMID- 12723676 TI - Shoulder motions during the golf swing in male amateur golfers. AB - STUDY DESIGN: Prospective descriptive biomechanical analysis of shoulder motion in golf. OBJECTIVE: To characterize normal shoulder motion during the driving swing in male recreational golfers of various age groups. BACKGROUND: Shoulder trauma accounts for approximately 12% of all golf-related injuries. To design sport-specific rehabilitation programs for the injured golfer and exercise programs for the healthy golfer, clinicians and teachers need quantitative information describing range of motion requirements about the shoulder for the amateur player. METHOD AND MEASURE: Sixty-five male golfers were divided into 3 age groups: college, middle, and senior. A high-speed, 6-camera motion analysis system recorded 3-dimensional bilateral shoulder motion (vertical elevation, horizontal adduction, external rotation, and shoulder tum) for 3 swings of the driver. Group means for ranges and functional end points of motion were compared using a single-factor 1-way ANOVA (alpha = 0.05). RESULTS: All maximum values of shoulder motion were lower in the senior group than in the other 2 groups. At peak backswing, senior golfers exhibited 38 degrees less right-side shoulder external rotation than college golfers. However, from address, seniors horizontally abduct the right arm 18 degrees more than college golfers. In the older golfers, total range of motion was reduced for both shoulders in the vertical plane and for the left shoulder in the horizontal plane. CONCLUSIONS: This study describes shoulder motion for asymptomatic golfers of various age groups. These data may serve as a baseline reference for assessing disease- or injury-related changes in the golf swing and for designing sport-specific exercise and rehabilitation programs. PMID- 12723677 TI - Decreasing landing forces in children: the effect of instructions. AB - STUDY DESIGN: Randomized controlled experimental design. OBJECTIVES: To examine the effects of verbal instructions related to control of joint kinematics and auditory feedback on lowering the vertical peak ground reaction forces associated with landing from a jump in children. BACKGROUND: The majority of children's injuries are associated with activities that involve jumping and landing. Instructions related to joint motion may assist children in decreasing landing forces and thus reduce the chances of sustaining an injury. METHODS AND MEASURES: Sixty-one school children whose mean (+/- SD) age was 9 (+/- 0.89) years participated. The children were randomly assigned (by school) to either experimental (received instruction on lower limb position and auditory cues) or control (received no instruction) groups. Participants were required to step off a box 300 mm in height and land on a force plate. The testing procedure involved 5 sessions. The first 4 sessions were performed over 3 days in the same week, while session 5 was performed approximately 3 months later to assess retention of the skill. The main outcome measure was the peak vertical ground reaction force recorded at foot strike. RESULTS: After instructions, subjects in the experimental group produced lower peak ground reaction forces (P<0.05) during landing compared to their control counterparts (ie, sessions 2-4). There was no significant difference (P<0.05) between groups at session 5 (3 months postinstruction). CONCLUSIONS: Children are capable of assimilating precise instruction to minimize forces on the lower limbs during landing from a jump. Providing further opportunity (ie, more than 1 session) to perform the skill does not alter the magnitude of the effect observed after the initial session of instruction. However, the improvements in landing performance were temporary. PMID- 12723678 TI - Relation between structure, function, and imaging in a three-dimensional model of the lung. AB - Previous studies have reported morphometric models to predict function relations in the lung. These models, however, are not anatomically explicit. We have advanced a three-dimensional airway tree model to relate dynamic lung function to alterations in structure, particularly when constriction patterns are imposed heterogeneously inspecific anatomic locations. First we predicted the sensitivity of dynamic lung resistance and elastance (RL and EL) to explicit forms of potential constriction patterns. Simulations show that severe and heterogeneous peripheral airway constriction confined to a single region in the lung (apex, mid, or base) will not produce substantial alterations in whole lung properties as measured from the airway opening. Conversely, when measured RL and EL are abnormal, it is likely that significant (but not necessarily homogeneous) constriction has occurred throughout the entire airway tree. We also introduce the concept of image-assisted modeling. Here positron emission tomographic imaging data sensitive to ventilation heterogeneity is synthesized with RL and EL data to help identify which airway constriction conditions could be consistent with both data sets. An ultimate goal would be personalized predictions. PMID- 12723679 TI - Computational fluid dynamics simulation of airflow and aerosol deposition in human lungs. AB - Computational fluid dynamics (CFD) simulations of airflow and particle deposition in geometries representing the human tracheobronchial tree were conducted. Two geometries were used in this work: (1) based on the Weibel A model, and (2) based on a CT scan of a cadaver lung cast. Flow conditions used included both steady state inhalation and exhalation conditions as well as time-dependent breathing cycles. Particle trajectories were calculated in each of these models by solving the equations of motion of the particle for the deterministic portion of particle displacement, and adding a stochastic Brownian term at each step. The trapping of particles on the wall surfaces was monitored, and the locations of trapping in each generation were recorded. The results indicate that there are dramatic differences in the predicted deposition between the two models. The intragenerational deposition locations show that in regions where the deposition mechanism is inertial impaction, the predominant deposition seems to be at the airway bifurcations. The results of this study suggest that under most conditions, an idealized model based on the Weibel dimensions is not sufficient to predict deposition, and an accurate model, such as those based on imaging techniques may be required. PMID- 12723680 TI - Phenotype modulation in vascular tissue engineering using biochemical and mechanical stimulation. AB - Biochemical stimulation was applied in combination with cyclic mechanical strain to engineered vascular constructs made of isolated smooth muscle cells in a three dimensional (3D) collagen type 1 matrix. Platelet-derived growth factor (PDGF) and transforming growth factor beta (TGF-beta) were added exogenously to the medium used to culture the constructs. Mechanical stimulation was applied using a bioreactor system that imparted a 10% circumferential strain at a frequency of 1 Hz. The parameters studied were gel compaction, cell proliferation, and expression of the contractile protein smooth muscle alpha-actin (SMA). Mechanical stimulation caused a characteristic increase in gel compaction and cell proliferation, relative to statically cultured controls. Stimulation with PDGF increased cell proliferation and decreased SMA expression in 3D gels, but inhibited the effects of mechanical stimulation and produced a more open matrix structure. TGF-beta strongly inhibited cell proliferation and increased SMA expression, especially in the presence of mechanical strain, and resulted in a dense matrix. These results show that cell phenotype can be modulated in engineered blood vessels by applying selected combinations of biochemical and mechanical stimuli, and suggest that such control over cell function can be used to tailor the properties of engineered tissues. PMID- 12723682 TI - Modeling the adaptive permeability response of porcine iliac arteries to acute changes in mural shear. AB - The hypothesis that much of the uptake of macromolecules by the vascular wall takes place while the endothelial lining is adapting to changes in its hemodynamic environment is being tested by a series of in vivo measurements of the uptake of Evans-blue-dye-labeled albumin by porcine iliac arteries subjected to acute changes in blood flow. The uptake data are interpreted through an ad hoc model of the dynamic permeability response that is proposed to accompany alterations in mural shear. The model is able to correlate, with a single set of parameters, the vascular response to a variety of experimental protocols, including sustained step increases and decreases in shear, and alternations in shear of various periods. The best-fit parameters of the model suggest that the adaptive response to an increase in shear proceeds with a latency of approximately 1.5 min and a time constant of approximately 90 min that is substantially shorter than the response to a decrease in shear. PMID- 12723681 TI - Arterial wall adaptation under elevated longitudinal stretch in organ culture. AB - Arteries in vivo are subjected to large longitudinal stretch which may change significantly due to vascular disease and surgery. However, little is known about the effect of longitudinal stretch on vascular function and wall remodeling, although the effects of tensile and shear stress from blood pressure and flow have been well documented. To study the effect of longitudinal stretch on vascular function and wall remodeling, porcine carotid arteries were longitudinally stretched 20% more than in vivo for 5 days while being maintained in an ex vivo organ culture system under conditions of pulsatile flow at physiologic pressure. Vessel viability was demonstrated by strong vasomotor responses to norepinephrine (NE, 10(-6) M), carbachol (10(-6) M), and sodium nitroprusside (10(-5) M), as well as by dense staining for mitochondrial activity and a low occurrence of cell necrosis. Cell proliferation was examined by incorporation of bromodeoxyuridine (BrdU). Results showed that arteries maintain normal structure and viability after 5 days in organ culture. Both the stretched and control arteries demonstrated significant contractile responses. For example, both stretched and control arteries showed approximately 10% diameter contraction in response to NE. Stretched arteries contained 8% BrdU-positive cells compared to 5% in controls (p<0.05). These results indicate that longitudinal stretch promotes cell proliferation in arteries while maintaining arterial function. PMID- 12723683 TI - Effects of cardiac motion on right coronary artery hemodynamics. AB - The purpose of this work was to investigate the effects of physiologically realistic cardiac-induced motion on hemodynamics in human right coronary arteries. The blood flow patterns were numerically simulated in a modeled right coronary artery (RCA) having a uniform circular cross section of 2.48 mm diam. Arterial motion was specified based on biplane cineangiograms, and incorporated physiologically realistic bending and torsion. Simulations were carried out with steady and pulsatile inflow conditions (mean ReD=233, alpha=1.82) in both fixed and moving RCA models, to evaluate the relative importance of RCA motion, flow pulsation, and the interaction between motion and flow pulsation. RCA motion with a steady inlet flow rate caused variations in wall shear stress (WSS) magnitude up to 150% of the inlet Poiseuille value. There was significant spatial variability in the magnitude of this motion-induced WSS variation. However, the time-averaged WSS distribution was similar to that predicted in a static model representing the time-averaged geometry. Furthermore, the effects of flow pulsatility dominated RCA motion-induced effects; specifically, there were only modest differences in the WSS history between simulations conducted in fixed and moving RCA models with pulsatile inflow. RCA motion has little effect on time averaged WSS patterns. It has a larger effect on the temporal variation of WSS, but even this effect is overshadowed by the variations in WSS due to flow pulsation. The hemodynamic effects of RCA motion can, therefore, be ignored as a first approximation in modeling studies. PMID- 12723685 TI - Modified magnetic resonance image based parcellation method for cerebral cortex using successive fuzzy clustering and boundary detection. AB - Development of the accurate and reproducible parcellation of the human brain can be used to resolve the complex structure-functional relationships in the brain. We propose a modified parcellation method that provides the reliable and reproducible regions of interest using successive fuzzy c-means (sFCM) and boundary-detection algorithm. This method displays simultaneously both original brain image for identifying the sulcal landmarks and its tissue-classified image for referring to patterns of sulci. The whole cerebral region is extracted by the semiautomated region growing method and then classified to gray matter, white matter, and cerebrospinal fluid by sFCM. Referred to the other previous researches, the volume ratio of gray matter to white matter was shown to find that the efficiency of classification was improved (conventional FCM: 0.80 +/- 0.12 vs. sFCM: 1.57 +/- 0.18). Inter-rater reliability, estimated by the regression analysis, demonstrated that the proposed method was more reliable and reproducible than conventional methods [ANALYZE: correlation coefficient (CC)=0.341, Sig.=0.335 vs. proposed method: CC=0.816, Sig.=0.004]. The volume ratio of the whole cerebrum to the parceled object can be used to investigate structural abnormalities for the pathological detection of the various mental diseases such as schizophrenia, obsessive-compulsive disorder. PMID- 12723684 TI - Novel technique for cardiac electromechanical mapping with magnetic resonance imaging tagging and an epicardial electrode sock. AB - Near-simultaneous measurements of electrical and mechanical activation over the entire ventricular surface are now possible using magnetic resonance imaging tagging and a multielectrode epicardial sock. This new electromechanical mapping technique is demonstrated in the ventricularly paced canine heart. A 128 electrode epicardial sock and pacing electrodes were placed on the hearts of four anesthetized dogs. In the magnetic resonance scanner, tagged cine images (8-15 ms/frame) and sock electrode recordings (1000 Hz) were acquired under right ventricular pacing and temporally referenced to the pacing stimulus. Electrical recordings were obtained during intermittent breaks in image acquisition, so that both data sets represented the same physiologic state. Since the electrodes were not visible in the images, electrode recordings and cine images were spatially registered with Gd-DTPA markers attached to the sock. Circumferential strain was calculated at locations corresponding to electrodes. For each electrode location, electrical and mechanical activation times were calculated and relationships between the two activation patterns were demonstrated. This method holds promise for improving understanding of the relationships between the patterns of electrical activation and contraction in the heart. PMID- 12723686 TI - A computational model of direct interstitial infusion of macromolecules into the spinal cord. AB - Convection-enhanced interstitial infusion can deliver macromolecular drugs to large tissue volumes of the central nervous system. To characterize infusion into the spinal cord, an image-based three-dimensional finite element model of the rat spinal cord was developed. The model incorporated convection and diffusion through white and gray matter, including anisotropic transport due to alignment of white matter tracts. Spatial and temporal distribution of the marker substance albumin within the interstitial space was determined. Consistent with previous experiments, predicted distribution was highly anisotropic. Infusing into the dorsal column, albumin was primarily confined to, white matter with limited penetration into adjacent gray matter. Distribution was determined primarily by the ratio of fiber-parallel to fiber-perpendicular hydraulic conductivity tensor components (k(wm-z)/k(wm-x)), the ratio of transverse white and gray matter hydraulic conductivity (k(wm-x)/k(gm)), and tissue porosity. Fits to previous experimental measures of axial and transverse spread, distribution volume, and protein recovery yielded an optimum k(wm-z)/k(wm-x) of approximately 20 at 0.1 microl/min. k(wm-x)/k(gm) of 100 was sufficient to match experimental transverse distribution data. Best fits to data at 0.1 microl/min were achieved by porosities characteristic of moderate edema (e.g., 0.26). Distribution also varied with catheter placement with more medial placement resulting in greater distribution volumes. PMID- 12723687 TI - Recognition of adenosine triphosphate binding sites using parallel cascade system identification. AB - Parallel cascade identification (PCI) is a method for approximating the behavior of a nonlinear system, from input/output training data, by constructing a parallel array of cascaded dynamic linear and static nonlinear elements. PCI has previously been shown to provide an effective means for classifying protein sequences into structure/function families. In the present study, PCI is used to distinguish proteins that are binding to adenosine triphosphate or guanine triphosphate molecules from those that are nonbinding. Classification accuracy of 87.1% using the hydrophobicity scale of Rose et al. (Hydrophobicity of amino acid residues in globular proteins. Science 229:834-838, 1985), and 88.8% using Korenberg's SARAH1 scale, are obtained, as measured by tenfold cross-validation testing. Nearest-neighbor and K-nearest-neighbor (KNN) classifiers are constructed, and the resulting accuracy is, respectively, 88.0% and 90.8% on the SARAH1-encoded test data set, as measured by the above testing protocol. Significantly improved classification accuracy is achieved by combining PCI and KNN classifiers using quadratic discriminant analysis: accuracy rises from 87.9% (PCI) and 87.4% (KNN) to 96.5% for the combination, as measured by twofold cross validation testing on the SARAH1-encoded test data set. PMID- 12723688 TI - Mathematical model of diffusion-limited evolution of multiple gas bubbles in tissue. AB - Models of gas bubble dynamics employed in probabilistic analyses of decompression sickness incidence in man must be theoretically consistent and simple, if they are to yield useful results without requiring excessive computations. They are generally formulated in terms of ordinary differential equations that describe diffusion-limited gas exchange between a gas bubble and the extravascular tissue surrounding it. In our previous model (Ann. Biomed. Eng. 30: 232-246, 2002), we showed that with appropriate representation of sink pressures to account for gas loss or gain due to heterogeneous blood perfusion in the unstirred diffusion region around the bubble, diffusion-limited bubble growth in a tissue of finite volume can be simulated without postulating a boundary layer across which gas flux is discontinuous. However, interactions between two or more bubbles caused by competition for available gas cannot be considered in this model, because the diffusion region has a fixed volume with zero gas flux at its outer boundary. The present work extends the previous model to accommodate interactions among multiple bubbles by allowing the diffusion region volume of each bubble to vary during bubble evolution. For given decompression and tissue volume, bubble growth is sustained only if the bubble number density is below a certain maximum. PMID- 12723689 TI - Effect of postmortem changes and freezing on the viscoelastic properties of vocal fold tissues. AB - It is common practice in laryngeal research laboratories to store excised larynges and vocal fold tissue specimens frozen, presumably to preserve viability and structural integrity of the specimens for physiologic and biomechanical experiments. However, little is known about the biomechanical effects of postmortem changes and frozen storage in vocal fold tissues. This study attempted to quantify the effects of postmortem changes and freezing on the viscoelastic shear properties of the canine vocal fold mucosa. Sixteen larynges were excised from adult dogs immediately postmortem in a viable state. Using a torsional rheometer, the complex shear modulus (G*) of the mucosal tissue from one vocal fold of each larynx was measured as a function of frequency (0.01-15 Hz) by linear small-amplitude oscillation experiments. Measurement was repeated for ten mucosal specimens after 24 h of postmortem storage in saline solution at room temperature. Eleven of the 16 larynges were frozen and stored at -20 degrees C, six of them at a slow rate of cooling (storage in a regular freezer) and five others by quick freezing (chilling by liquid nitrogen prior to frozen storage). The larynges were thawed slowly overnight after one month and the viscoelastic shear properties of the contralateral vocal fold mucosa were measured. Results showed that the elastic shear modulus (G') and dynamic viscosity (eta') of the vocal fold mucosa did not seem to change significantly after 24 h of storage in saline at room temperature, nor after one month of frozen storage following quick freezing, whereas both G' and eta' decreased significantly for tissues that were slowly frozen. These findings supported the feasibility of using quick freezing to preserve laryngeal tissues for in vitro biomechanical testing, for excised larynx experiments, and for tissue engineering applications. PMID- 12723690 TI - Expanding the boundaries of heterochromatin. PMID- 12723692 TI - HP1/ORC complex and heterochromatin assembly. AB - We have used the highly conserved heterochromatin component, heterochromatin protein 1 (HP1), as a molecular tag for purifying other protein components of Drosophila heterochromatin. A complex of HP1 associated with the origin recognition complex (ORC) and an HP1/ORC-associated protein (HOAP) was purified from the maternally loaded cytoplasm of early Drosophila embryo. We propose that the DNA-binding activities of ORC and HOAP function to recruit underphosphorylatedisoforms of HP1 to sites of heterochromatin nucleation. The roles of highly phosphorylated HP1, other DNA-binding proteins known to interact with HP1, and histone modifying activities in heterochromatin assembly are also addressed. PMID- 12723691 TI - Analysing the contribution of nucleic acids to the structure and properties of centric heterochromatin. AB - A class of repetitive DNA sequences frequently found at centromeric regions are R/Y-satellites showing an asymmetric distribution of residues resulting in one strand being rich in purines (R-strand) while the complementary strand is pyrimidine-rich (Y-strand). The dodeca-satellite of Drosophila belongs to this class of centromeric satellites. In vitro, the dodeca-satellite forms altered DNA structures in which the R-strand forms very stable intramolecular fold-backs that are stabilised by the formation of tandem G x A mismatches. A single-stranded nucleic acids binding protein, DDP1, binds the unstructured dodeca-satellite Y strand with high affinity. In polytene chromosomes, DDP1 associates with the heterochromatic chromocenter and, at the euchromatic chromosome arms, co localises with HP1. DDP1 is a vigilin. Vigilins are highly conserved multi-KH domain proteins. Scp160p, the vigilin from S. cerevisiae, is involved in the control of ploidy. DDP1 complements a deltascp160 deletion. PMID- 12723693 TI - Chromosomal distribution of heterochromatin protein 1 (HP1) in Drosophila: a cytological map of euchromatic HP1 binding sites. AB - The Heterochromatin Protein 1 (HP1) is a conserved protein which is best known for its strong association with the heterochromatin of Drosophila melanogaster. We previously demonstrated that another important property of HP1 is its localization to the telomeres of Drosophila, a feature that reflects its critical function as a telomere capping protein. Here we report our analysis of the euchromatic sites to which HP1 localizes. Using an anti-HP1 antibody, we compared immunostaining patterns on polytene chromosomes of the Ore-R wild type laboratory strain and four different natural populations. HP1 was found to accumulate at specific euchromatic sites, with a subset of the sites conserved among strains. These sites do not appear to be defined by an enrichment of known repetitive DNAs. Comparisons of HP1 patterns among several Drosophila species revealed that association with specific euchromatic regions, heterochromatin and telomeres is a conserved characteristic of HP1. Based on these results, we argue that HP1 serves a broader function than typically postulated. In addition to its role in heterochromatin assembly and telomere stability, we propose that HP1 plays an important role in regulating the expression of many different euchromatic regions. PMID- 12723694 TI - SU(VAR)3-9 is a conserved key function in heterochromatic gene silencing. AB - This review summarizes genetic, molecular and biochemical studies of the SU(VAR)3 9 protein and the evidence for its key role in heterochromatin formation and heterochromatic gene silencing. The Su(var)3-9 locus was first identified as a dominant modifier of position-effect variegation (PEV) in Drosophila melanogaster. Together with Su(var)2-5 and Su(var)3-7, Su(var)3-9 belongs to the group of haplo-suppressor loci which show a triplo-dependent enhancer effect. All three genes encode heterochromatin-associated proteins. Su(var)3-9 is epistatic to the PEV modifier effects of Su(var)2-5 and Su(var)3-7, and it also dominates the effect of the Y chromosome on PEV. These genetic data support a central role of the SU(VAR)3-9 protein in heterochromatic gene silencing, one that is correlated with its activity as a histone H3-K9 methyltransferase (HMTase). In fact, SU(VAR)3-9 is the main chromocenter-specific HMTase of Drosophila. SU(VAR)3 9 and HP1, the product of Su(var)2-5, are main constituents of heterochromatin protein complexes and the interaction between these two proteins is interdependent. Functional analysis in fission yeast, Drosophila and mammals demonstrate that SU(VAR)3-9-dependent gene silencing processes are conserved in these organisms. This is also demonstrated by the rescue of Drosophila Su(var)3-9 mutant phenotypes with human SUV39H1 transgenes. PMID- 12723695 TI - The sounds of silence--histone deacetylation meets histone methylation. AB - The repression of gene activity and the maintenance of the repressed state are fundamental requirements of cell differentiation, ordered embryonic development and tissue integrity. Furthermore, large regions of the genome such as centromeres and telomeres have a structural function and have to be kept transcriptionally inactive to be functional. In both cases the transcriptional silencing is accomplished through a dense packaging of the corresponding DNA into heterochromatin or heterochromatin-like structures. In this minireview we summarise recent findings, which point towards a major function of posttranslational histone modifications in the process of establishment and maintenance of condensed heterochromatin. The physical association of two enzymatic activities, histone methylation and histone deacetylation, which are thought to be involved in transcriptional silencing, provide the framework of a molecular model of how heterochromatin is initiated and maintained during cell division and differentiation. PMID- 12723696 TI - The modifier of mdg4 locus in Drosophila: functional complexity is resolved by trans splicing. AB - The modifier of mdg4 (mod(mdg4)) gene of Drosophila melanogaster has been identified in many different genetic assays. It has been independently identified through mutations isolated for their effects on position effect variegation (PEV), the properties of insulator sequences, correct pathfinding of growing nerve cells, meiotic pairing of chromosomes, or apoptosis. Molecular analysis of the mod(mdg4) locus revealed that it encodes a family of at least 26 protein isoforms. Inspired by the fact that some mod(mdg4) transcripts are encoded by both antiparallel DNA strands, it was shown that mRNA trans splicing is the mechanism used by this locus to produce mature transcripts. All Mod(mdg4) protein isoforms share a common N-terminal region of 402 amino acids, which includes the conserved BTB/POZ domain. However, the isoforms differ in their C-terminal ends. Most of the C-termini contain a conserved Cys2His2 protein motif, which we have named the FLYWCH motif. Genetic and immunological data indicate that mod(mdg4) encodes a family of related chromatin proteins. Recent results indicate a functional correlation between the large number of different isoforms and the pleiotropic mutant phenotypes of most mod(mdg4) mutations. We discuss the putative function of Mod(mdg4) proteins as chromatin modulators involved in higher order chromatin domains. We also provide evidence for the evolutionary conservation of several of the isoforms and the unusual structure of the locus. PMID- 12723697 TI - Dosage dependent gene regulation and the compensation of the X chromosome in Drosophila males. AB - Dosage compensation refers to the phenomenon that despite the difference in copy number of X chromosomes in males and females approximately equal expression results from the sex chromosomes. We describe evidence for a model that dosage compensation is caused by an 'inverse dosage effect' that results from an altered stoichiometry of transcriptional regulators on the X versus the autosomes. This imbalance of regulators would cause a two-fold increase in target gene expression throughout the genome in the absence of any modification. The two-fold hyperactivation compensates the X chromosome. However, the MSL (male specific lethal) complex modifies this effect on the autosomes, which would otherwise double their expression, by becoming sequestered to the X chromosome together with a histone acetylase (MOF) and kinase (JIL1). This situation reduces the level of histone 4 Lys16 acetylation and H3 phosphorylation on the autosomes, thus bringing their expression down to near the female level. The presence of the MSL complex on the X modifies chromatin in such a manner that it counteracts any impact of increased histone acetylation and phosphorylation on gene expression. This situation fosters the proper two-fold increase in gene expression needed for X chromosomal dosage compensation in males and reduces the inverse effect on the autosomes to equalize gene expression throughout the genome for the two sexes. PMID- 12723698 TI - Assembly of Polycomb complexes and silencing mechanisms. AB - Polycomb complexes assemble at their target sites and silence neighboring genes when these are not actively transcribed. The action of these complexes and of Trithorax complexes bound to the Polycomb Response Element establish alternative silent or derepressed states that are remembered through cell division and maintained for the rest of development. Recent results that may help explain the properties of these states are reviewed. PMID- 12723699 TI - Composite cis-acting epigenetic switches in eukaryotes: lessons from Drosophila Fab-7 for the Igf2-H19 imprinted domain. AB - One of the central problems of eukaryotic gene regulation is to understand the mechanism(s) by which the activity of enhancer elements is circumscribed such that they only act upon their cognate promoter sequences. Studies on the bithorax gene complex (BX-C) in Drosophila have highlighted the potential problem of enhancer promiscuity and detailed molecular and genetic analyses are now providing insight into how this gene complex resolves the problem through the activity of boundary/silencer elements that can block the communication between enhancers and promoters. Analysis of the mouse Igf2-H19 imprinted locus also suggests a role for boundary/silencer elements, but in this case these elements are invoked to account for the preferential expression of Igf2 and H19 from the paternally and maternally inherited chromosomes respectively despite the presence of functional downstream enhancers. We discuss recent work that has illuminated both of these systems and consider what parallels exist between them. PMID- 12723700 TI - Vital genes in the heterochromatin of chromosomes 2 and 3 of Drosophila melanogaster. AB - Heterochromatin has been traditionally regarded as a genomic wasteland, but in the last three decades extensive genetic and molecular studies have shown that this ubiquitous component of eukaryotic chromosomes may perform important biological functions. In D. melanogaster, about 30 genes that are essential for viability and/or fertility have been mapped to the heterochromatin of the major autosomes. Thus far, the known essential genes exhibit a peculiar molecular organization. They consist of single-copy exons, while their introns are comprised mainly of degenerate transposons. Moreover, about one hundred predicted genes that escaped previous genetic analyses have been associated with the proximal regions of chromosome arms but it remains to be determined how many of these genes are actually located within the heterochromatin. In this overview, we present available data on the mapping, molecular organization and function of known vital genes embedded in the heterochromatin of chromosomes 2 and 3. Repetitive loci, such as Responder and the ABO elements, which are also located in the heterochromatin of chromosome 2, are not discussed here because they have been reviewed in detail elsewhere. PMID- 12723701 TI - A strategy for mapping the heterochromatin of chromosome 2 of Drosophila melanogaster. AB - The heterochromatin of chromosome 2 of Drosophila melanogaster has been among the best characterized models for functional studies of heterochromatin owing to its abundance of genetic markers. To determine whether it might also provide a favorable system for mapping extended regions of heterochromatin, we undertook a project to molecularly map the heterochromatin of the left arm of chromosome 2 (2Lh). In this paper, we describe a strategy that used clones and sequence information available from the Drosophila Genome Project and chromosome rearrangements to construct a map of the distal most portion of 2Lh. We also describe studies that used fluorescent in situ hybridization (FISH) to examine the resolution of this technique for cytologically resolving heterochromatic sequences on mitotic chromosomes. We discuss how these mapping studies can be extended to more proximal regions of the heterochromatin to determine the structural patterns and physical dimensions of 2Lh and the relationship of structure to function. PMID- 12723702 TI - Y chromosome and other heterochromatic sequences of the Drosophila melanogaster genome: how far can we go? AB - Whole genome shotgun assemblies have proven remarkably successful in reconstructing the bulk of euchromatic genes, with the only limit appearing to be determined by the sequencing depth. For genes imbedded in heterochromatin, however, the low cloning efficiency of repetitive sequences, combined with the computational challenges, demand that additional clues be used to annotate the sequences. One approach that has proven very successful in identifying protein coding genes in Y-linked heterochromatin of Drosophila melanogaster has been to make a BLASTable database of the small, unmapped contigs and fragments leftover at the end of a shotgun assembly, and to attempt to capture these by blasting with an appropriate query sequence. This approach often yields a staggered alignment of contigs from the unmapped set to the query sequence, as though the disjoint contigs represent small portions of the gene. Further inspection frequently shows that the contigs are broken by very large, heterochromatic introns. Methods of this sort are being expanded to make best use of all available clues to determine which unmapped contigs are associated with genes. These include use of EST libraries, and, in the case of the Y chromosome, testing of male specific genes and reduced shotgun depth of relevant contigs. It appears much more hopeful than anyone would have imagined that whole genome shotgun assemblies can recover the great bulk of even heterochromatic genes. PMID- 12723703 TI - Stellate repeats: targets of silencing and modules causing cis-inactivation and trans-activation. AB - The mechanism of silencing of testis expressed X-linked Stellate repeats by homologous Y-linked Suppressor of Stellate [Su(Ste)] repeats localized in the crystal locus was studied. The double stranded RNA as a product of symmetrical transcription of Su(Ste) repeat and small interference Su(Ste) siRNA were revealed suggesting the mechanism of RNA interference (RNAi) for Stellate silencing. The relief of Stellate silencing as a result of impaired complementarity between the sequences of putative target Stellate transcripts and Su(Ste) repeats was shown. The role of RNAi mechanism in the silencing of heterochromatic retrotransposon GATE inserted in Stellate cluster was revealed. The studies of cis-effects of Stellate tandem repeats causing variegated expression of juxtaposed reporter genes were extended and the lacZ variegation in imaginal disc was shown. The exceptional case of a non-variegated expression of mini-white gene juxtaposed to Stellate repeats in a construct inserted into the 39C region was shown to be accompanied by trans-activation in homozygous state. Trans-activation effect was retained after transposition of this construct into heterochromatic environment in spite of strong variegation of a mini-white gene. PMID- 12723704 TI - Structure, regulation and evolution of the crystal-Stellate system of Drosophila. AB - The crystal-Stellate system is one of the most known example of interaction between heterochromatin and euchromatin: a heterochromatic locus on the Y chromosome (crystal) 'represses' a euchromatic locus (Stellate) on the X chromosome in Drosophila melanogaster. The molecular mechanism regulating this interaction is not completely understood. It is becoming clear that an RNA interference (RNAi) mechanism could be responsible for the silencing carried out by crystal on the Stellate sequences. Here, a detailed structural analysis of all the sequences involved in the system is reported, demonstrating a their 'puzzling' structure. In addition three autosomal mutations: sting, scratch and sirio are described that interfere with the system. All of them are male sterile mutations and exhibit crystals made by the STELLATE protein in their primary spermatocytes. They are requested during oogenesis and early in embryogenesis as well. Hypothesis on the involvement of these genes in activating the Stellate sequences are discussed. PMID- 12723705 TI - Intercalary heterochromatin in Drosophila melanogaster polytene chromosomes and the problem of genetic silencing. AB - The morphological characteristics of intercalary heterochromatin (IH) are compared with those of other types of silenced chromatin in the Drosophila melanogaster genome: pericentric heterochromatin (PH) and regions subject to position effect variegation (PEV). We conclude that IH regions in polytene chromosomes are binding sites of silencing complexes such as PcG complexes and of SuUR protein. Binding of these proteins results in the appearance of condensed chromatin and late replication of DNA, which in turn may result in DNA underreplication. IH and PH as well as regions subject to PEV have in common the condensed chromatin appearance, the localization of specific proteins, late replication, underreplication in polytene chromosomes, and ectopic pairing. PMID- 12723706 TI - Imprinted facultative heterochromatization in mealybugs. AB - In lecanoid Coccids, or mealybugs, the male development is accompanied by the facultative heterochromatization of the entire, paternally derived, haploid chromosome set. This epigenetic phenomenon occurs in all the cells of mid cleavage male embryos. Consequently, the Coccid chromosome system offers a powerful tool for gaining insights into the structure of facultative heterochromatin, and into the epigenetic mechanisms of its imprinted, developmentally regulated formation. This paper will present new data and summarize recent studies on genomic imprinting and facultative heterochromatization in mealybugs. First, the existence and the possible role of DNA methylation as an epigenetic modification that fulfills the requisites of the imprinting process in mealybugs will be considered. The second part of this paper will focus on proteins involved in the facultative heterochromatization process. In particular, the involvement of an HP-1-like protein in the silencing of the paternally derived haploid chromosome set and its interaction with the lysine 9 methylated isoform of histone H3 will be discussed. PMID- 12723707 TI - Organization and possible origin of the Bari-1 cluster in the heterochromatic h39 region of Drosophila melanogaster. AB - The molecular organization of the heterochromatic h39 region of the Drosophila melanogaster second chromosome has been investigated by studying two BAC clones identified both by Southern blotting and by FISH experiments as containing tandem arrays of Bari1, a transposable element present only in this region. Such BAC clones appear to contain different portions of the h39 region since they differ in the DNA sequences flanking the Bari1 repeats on both sides. Thus, the 80 Bari1 copies estimated to be present in the h39 region are split into at least two separated subregions. On the basis of the analysis of the flanking sequences a possible mechanism depending on an aberrant activity of the Bari1 transposase is proposed for the genesis of the heterochromatic tandem arrays of the element. PMID- 12723708 TI - Responder (Rsp) alleles in the segregation distorter (SD) system of meiotic drive in Drosophila may represent a complex family of satellite repeat sequences. AB - In D. melanogaster males carrying Segregation Distorter (SD) second chromosomes, sperm receiving sensitive alleles of the Responder (Rsp) locus are subject to high rates of dysfunction. The Rsp region is located in 2R immediately adjacent to the centromere in heterochromatic band 39, and covers roughly 600 kb of material, of which approximately 85 kb is comprised of several hundred copies of a 240-bp satellite DNA sequence. Cytological observations as well as molecular analysis of rearrangements which bisect h39 indicate that sensitivity of the Rsp target to SD action is also subdivisible, and sensitivities of the component pieces appear to be correlated with copy number of the 240 bp repeat. In an attempt to examine possible higher order sequence structure for these blocks, PCR using single primers derived from a canonical repeat was used to identify potential reversals of direction of tandem arrays; that is, head-to-head or tail to-tail junctions. Surprisingly, for two different Rsp alleles, only a single such reversal product for each was identified, differing in size and sequence between alleles. Sequencing of PCR products identified diverged copies of the canonical repeats that would not have been found using the levels of DNA stringency employed in earlier studies. Examination of Southern digests and slot blots for DNA quantification indicates that adding the estimated numbers of such diverged copies to the canonical repeat copies discovered earlier is potentially sufficient to account for the entire 600 kb Rsp region. This adds strength to the hypothesis that this extended family of repeats is in fact the target of SD mediated sperm dysfunction. Implications of these results for understanding the evolution of repetitive DNA are also discussed. PMID- 12723709 TI - A subset of the elements of the 1731 retrotransposon family are preferentially located in regions of the Y chromosome that are polytenized in larval salivary glands of Drosophila melanogaster. AB - It has been previously reported that the abundance and distribution of transposable elements (TEs) in Drosophila heterochromatin are conserved in unrelated stocks although they may greatly differ between families. The biases in genomic distribution of TEs are potentially informative for understanding host transposon interactions. Here we report that in most stocks, one to four elements of the 1731 retrotransposon family are located on the Y chromosome within regions that appear to be polytenized in larval salivary glands. We discuss the hypothesis that these elements may be beneficial to the host and consider the relevance of our observations to the organization of sequences within the heterochromatin. PMID- 12723710 TI - The role of HeT-A and TART retrotransposons in Drosophila telomere capping. AB - Drosophila telomeres contain multiple copies of HeT-A and TART retrotransposons. These elements specifically transpose to chromosomal ends, compensating for loss of terminal nucleotides that occurs at each cycle of DNA replication. We have investigated the role of these sequences in the formation of telomere-telomere attachments induced by mutations in the UbcD1 gene. We have constructed UbcD1 mutant males carrying terminally deleted X chromosomes devoid of both HeT-A and TART sequences. Cytological analysis of larval neuroblasts from these males revealed that telomeres lacking HeT-A and TART and normal telomeres that contain these sequences participate in telomeric fusions with comparable frequencies. These results indicate that the UbcD1 substrate(s) binds chromosomal termini in a sequence-independent manner. Previous studies have shown that the telomere capping protein HP1 also binds telomeres lacking HeT-A and TART. Taken together, these findings strongly suggest that the assembly of DNA-protein complexes that protect chromosome ends from fusions do not require specific terminal sequences. PMID- 12723711 TI - Telomeric position effect in drosophila melanogaster reflects a telomere length control mechanism. AB - The terminal DNA arrays on chromosomes of Drosophila melanogaster are composed of two families of non-LTR retrotransposons, HeT-A and TART. Available evidence suggests that chromosome length in this species and its close relatives is maintained by targeted transposition of these elements, with attachment of the elements to the chromosome end by their 3' oligo(A) tails. However, the regulation of transposition of these elements and the control of telomere length are poorly understood. Here we present the hypothesis that the forces involved in telomere length regulation in Drosophila are the underlying forces that manifest themselves as telomeric position effect (TPE). Based on recent studies of TPE, which found that expression of a reporter gene is influenced by telomere structure in cis and trans, we propose that the subtelomeric satellite (TAS) in D. melanogaster plays an important role in controlling telomere elongation. Transcription of a HeT-A element is probably initiated at a promoter in the 3' UTR of an upstream element, and TAS may repress this transcriptional activity in cis and trans. A region of HeT-A not at the extreme 3' end of the element may act as a transcriptional enhancer that may be modulated by TAS. PMID- 12723713 TI - PDE5 inhibitors: looking beyond ED. PMID- 12723712 TI - Telomeric transgenes and trans-silencing in Drosophila. AB - Autonomous P elements, inserted in heterochromatic telomeric associated sequences (TAS) at the X chromosome telomere (site 1A) have strong P element regulatory properties that include repression of P-induced hybrid-dysgenesis and of P-lacZ expression in the germline. P-lacZ insertions or defective P elements at 1A in TAS can also repress in trans a euchromatic P-lacZ in the germline. This property has been called a trans-silencing effect (TSE). It requires some sequence homology between the telomeric insertion and the euchromatic transgene. When repression is partial, variegating lacZ expression is observed, suggesting a chromatin-based component. TSE is observed only when the silencer transgenes are maternally inherited and occurs only in the female germline. We have evidence that this silencing also works in the presence of homologous non-P element sequences suggesting that homology-dependent silencing could be a general phenomenon in the female germline; such a system might have been subsequently adopted by the P element family, allowing its own repression. PMID- 12723714 TI - Vaccination against hepatitis B in patients on chronic haemodialysis. PMID- 12723715 TI - A simple questionnaire to detect hypermobility: an adjunct to the assessment of patients with diffuse musculoskeletal pain. AB - The aim of the study was to develop a simple and reproducible self-reporting questionnaire that identifies individuals with hypermobility. Two hundred and twelve consecutive hypermobile female new attendees to the hypermobility clinic at two London teaching hospitals and a random selection of 57 healthy volunteers completed a 10-part questionnaire. Questions were selected from clinical experience (RG), and assessed musculoskeletal symptoms and past and present physical agility. Of the 212 cases, 30 were hypermobile with no other underlying disorder and 182 fulfilled the 1998 Brighton criteria for benign joint hypermobility syndrome (BJHS). Odds ratios for the presence of hypermobility were calculated for each question. Six questions were found to be significant and the model of 'best fit' for sensitivity and specificity contained five of these. To demonstrate the reproducibility of the five-part questionnaire a second cohort of 170 hypermobile cases with BJHS and 50 controls was surveyed. Analysis demonstrated that a positive answer to any two questions in the five-part questionnaire gave the highest combined sensitivity and specificity for detecting hypermobility. The sensitivity and specificity was 84% and 89% respectively in the first cohort and reproduced with values of 84% and 80% in the second cohort. Overall the questionnaire correctly identified 84% of all cases and controls. This simple and reproducible questionnaire for detecting hypermobility could be of particular use as an adjunct in the clinical assessment of chronic, diffuse pain syndromes where hypermobility is often missed yet is potentially treatable. PMID- 12723716 TI - Why do family doctors prescribe antibiotics for upper respiratory tract infection? AB - The prescribing behaviour of family doctors in Hong Kong towards upper respiratory tract infections (URTIs), and the major clinical factors that might affect such behaviour, were studied. Members of the Hong Kong College of Family Physicians were surveyed; 801 completed the questionnaire with an overall response rate of 65%. Purulent nasal discharge, purulent sputum, persistent fever over three days, patients looking unwell, exudates on throat, inflamed eardrums and cervical lymphadenopathy made more than half the respondents 'likely' or 'very likely' to prescribe antibiotics. Those in private practice and those who graduated in Hong Kong were more likely to prescribe antibiotics, while fellows of the College were less likely to do so. Vocational training and higher qualifications in family medicine/general practice, however, revealed minimal effect. Our results showed that many doctors are still prescribing antibiotics when they encounter URTI patients presenting with clinical factors that have been proven to have no or little benefit from antibiotics. PMID- 12723717 TI - Growth charts for children with Down's syndrome in Saudi Arabia: birth to 5 years. AB - The objective was to construct growth charts for Saudi children under 5 years of age with Down's syndrome (DS). A prospective cross-sectional survey of anthropometrics standards (weight, height, head circumference and weight for height) of Saudi children with DS, excluding those with associated pathology, and a group of normal children seen at King Khalid University Hospital, Riyadh, Saudi Arabia, during 2001 were studied. Both groups were compared to a reference population of American children documented by the National Center for Health Statistics/Center for Disease Control. A total of 785 children with DS and 989 normal children were studied. Growth charts for the different parameters in both sexes at each age were used to estimate seven centiles. The growth pattern for children with DS differed significantly from normal children and was characterised by short stature at most ages. Data indicated a high proportion of underweight children with DS in the first two years of life, but a clear tendency to be overweight by the age of 3 years; 54.5% of children with DS had a head circumference below -2 SD for age and sex. The results of this study accord with others indicating a genetic effect of DS on growth, and support the use of growth charts in conjunction with charts for normal children for assessing growth in DS in Saudi Arabia. PMID- 12723718 TI - An open-label, multicentre study to assess the safety and efficacy of a novel reflux suppressant (Gaviscon Advance) in the treatment of heartburn during pregnancy. AB - This study investigated the efficacy and safety of a novel reflux suppressant, Gaviscon Advance, in the treatment of heartburn during pregnancy. The study was an open-label, multicentre, phase IV study in general practice and antenatal clinics in the UK and Republic of South Africa. Pregnant women (< or = 38 weeks gestation; n=150) aged 18-40 years suffering from heartburn were instructed to take Gaviscon Advance 5-10 ml, as required, to relieve symptoms. The main outcome measures were the efficacy rating of the study medication by the investigator and women after four weeks using a five-point efficacy scale. After four weeks the investigators' and women's rating of efficacy was 'very good' or 'good' in 88% and 90% of women, respectively. Most women (57%, n=83) reported symptom relief within 10 minutes. Thus Gaviscon Advance effectively and rapidly treats heartburn during pregnancy. Its use during pregnancy presents no known significant safety concerns for mother or child. PMID- 12723719 TI - Use of nephrostomy tubes in ureteric obstruction from incurable malignancy. AB - Malignancy may produce ureteric obstruction. This obstruction may be relieved by inserting a nephrostomy tube. All 31 cases of malignant ureteric obstruction from a single institution were retrospectively analysed. The commonest indications for nephrostomy tube insertion were renal failure (87%) and flank pain with hydronephrosis (13%). The mean serum creatinine levels pre and post insertion were 481 and 170 micromol/l, representing significant improvement (p<0.01). The complication rate was 13%, relating to tube blockage or dislodgement. Survival of patients was significantly worse if their malignancy was not resectable or suitable for chemotherapy (p=0.01). However, incurable patients still survived for a mean of 232 days, and although 46% of this time was spent as an inpatient, it did not differ significantly from the inpatient rate of curable patients (31%, p=0.1). Nephrostomy tube insertion should be considered for all palliative cases of malignant ureteric obstruction. PMID- 12723720 TI - Incontinence in adult females with cystic fibrosis: a Northern Ireland survey. AB - The objectives of this survey were to establish the prevalence, onset, severity and impact of incontinence and attitudes towards the availability of advice and treatment in female adults with cystic fibrosis (CF) in Northern Ireland. All female patients (n=59) at the Northern Ireland Regional Adult CF Centre were posted a questionnaire on incontinence. Leakage of urine occurred in 14/46 respondents. Leakage of urine occurred when chest was bad in 8/14 patients and when chest was good or bad in 3/14 patients. Patients reported that their bladder problem affected their ability to perform airway clearance or cough (13/14), and exercise (4/14). The main reason given for the patients who did not seek help for their incontinence (10/14) was that they were too embarrassed. This study highlights that patients with CF are reluctant to seek treatment for incontinence despite the impact this condition can have. PMID- 12723721 TI - Bone metastasis as the first manifestation of lung cancer. AB - Bone metastasis usually occurs late in the course of the disease, but in some patients it is the first manifestation of lung cancer. To evaluate the characteristics of patients with bone metastasis as a first manifestation of lung cancer, the medical records of 1063 patients with lung cancer between 1976 and 2001 were reviewed retrospectively. Twenty-four (2.3%) lung cancer patients presented symptoms of bone metastasis as a first manifestation; 11 patients had bone metastasis as the only site of spread in lung cancer; 16 had adenocarcinoma; five had a primary lesion <30 mm, and nine had N0-1 disease. Patients with sole bone metastasis did not have a more favourable survival rate than patients with bone and other systemic metastases (p=0.2938). Whether metastasis is the first manifestation of lung cancer or not, the occurrence of bone metastasis generally means a poor prognosis. PMID- 12723722 TI - Management of locally advanced prostate cancer: a European consensus. AB - This report summarises the findings of a European Consensus Group review of current standards of care in locally advanced prostate cancer defined as (a) untreated cancer extending clinically beyond the prostatic capsule in patients with no evidence of lymph node invasion or distant metastases, and (b) residual disease remaining after local treatment with positive surgical margins, seminal vesicle invasion, persistent prostate-specific antigen (PSA) and/or secondary PSA relapse. There was no overall consensus as to the standard of care in clinically apparent locally advanced prostate cancer. It was agreed, however, that hormonal therapy (e.g. with a gonadotrophin releasing hormone analogue [GnRHa]) represents a valid treatment in these patients. Treatment practices and regimens vary considerably between European countries, but GnRHa is widely used, either alone or in combination with antiandrogens. Hormonal therapy alone is a valid option, though the optimal modality, timing and duration of treatment remain to be defined. Adjuvant therapy with a GnRHa has been shown to improve survival in patients undergoing external beam radiotherapy. It is a viable option after prostatectomy in patients with persistent or secondary relapsing PSA. It was determined that optimal treatment will be different according to PSA, clinical staging and Gleason score, and the treatment of locally advanced disease should be individually tailored after discussion between physician and patient. In many instances, patients prefer and expect some form of treatment in preference to watchful waiting. Treatment nomograms such as the Kattan nomograms provide precise, comprehensive and invaluable tools for everyday use and may be used to predict outcomes and guide treatment decisions. PMID- 12723723 TI - 21. The adjuvant treatment of breast cancer. AB - Risk of relapse of breast cancer depends largely on tumour features: size, grade and, particularly, lymph node status. Adjuvant systemic therapies reduce risk of relapse and death, as shown in several meta-analyses. Relative risk reductions are similar across different prognostic groups but the absolute benefits are greater in those with a worse prognosis. Combination chemotherapy is superior to single agent regimens but longer duration (>6 months) gives no additional benefit compared with six months of therapy. Benefits from adjuvant chemotherapy diminish with the age of the patient. CMF type regimens have been most widely used: anthracyclines can reduce risk of relapse by 12% (an absolute survival benefit of about 3%) but with greater toxicity. Early trials of taxoids look promising but benefit was seen only in those with ER- tumours. Tamoxifen is the most extensively used endocrine adjuvant therapy, and is associated with a reduction in risk of death of 26% independent of age or menopausal status but only in those with ER+ tumours. Aromatase inhibitors are effective only in postmenopausal women and, although the early results with anastrozole look promising, long-term efficacy and toxicity studies are presently lacking, so five years of tamoxifen is standard treatment for women with ER+ tumours. PMID- 12723724 TI - 22. Diagnostic delays in breast cancer and impact on survival. AB - There is always an interval between the first symptoms, time to diagnosis and start of treatment in women with symptomatic breast cancer. Delay may be due to the patient, her general practitioner or a false negative diagnosis in hospital. A systematic overview showed that delays of 3-6 months were clearly associated with increased tumour size, advance in disease stage and poorer long-term prognosis. Nevertheless, controversy persists regarding the impact of delay on survival. The conflicting results may be due to a difference in sample characteristics, differences in the delay interval studied or variations in the definition of delay. A major drawback of many studies was that the potential confounding effect of lead-time bias was not taken into account. One of the largest single institution studies, designed to control for the lead-time bias, confirmed that survival measured from both date of diagnosis and onset of patient's symptoms was worse in women with delays of >12 weeks. Within individual stages, longer delays had no adverse impact on survival. It is important to identify groups of patients at high risk of delay so that strategies can be developed and effectively targeted. PMID- 12723725 TI - Auscultatory percussion: an added dimension in physical diagnosis. AB - The combination of both auscultation and percussion in the diagnostic examination improves both auscultation and percussion and, in so doing, increases the skill of palpation. Auscultatory percussion (AP) is easy to learn, easy to use and requires little time to perform. The size of normal organs and the size of abnormal masses as determined by AP are usually similar to measurements determined by X-rays and CT scans. Abnormalities found by AP need to be evaluated and confirmed by other diagnostic procedures. AP brings increased precision to physical diagnosis and is felt to be a valuable addition to medical practice. PMID- 12723726 TI - Endothelial dysfunction and cardiovascular disease: the promise of blocking the renin-angiotensin system. AB - As a crucial regulator of vascular function the endothelium balances a complex range of actions. Accordingly, damage to the endothelium often precedes the development of clinically manifest vascular disease. This review surveys our current understanding of risk factors involved in causing endothelial damage and the effects of lifestyle changes and pharmacotherapy on the endothelium. Our developing understanding of the intricacies of endothelial function and the effects of risk factors may aid in optimising cardiovascular prevention as well as therapy. PMID- 12723727 TI - Galantamine: a review of its use in Alzheimer's disease and vascular dementia. AB - Dementia has the potential to become a major public health concern during this century due to increasing life expectancy and growth in the ageing population. The commonest types of dementia include Alzheimer's disease (AD), vascular dementia (VaD), and dementia with cerebrovascular components. Galantamine is the only acetylcholinesterase inhibitor that exhibits a dual mechanism of action- inhibition of acetylcholinesterase and nicotinic receptor modulation. Clinical studies demonstrate the efficacy and safety of galantamine in patients with AD, VaD, and AD with cerebrovascular components. Galantamine shows beneficial effects on cognition, global function, activities of daily living and behaviour. Adverse events observed with galantamine use are generally mild to moderate in severity, transient and gastrointestinal in nature. The dose of galantamine should be escalated to 16 and 24 mg/day at four-week intervals to achieve maximal tolerability. Because of its unique mechanism of action, galantamine may have potential benefits over conventional enzyme-inhibiting agents. PMID- 12723728 TI - Efficacy and safety of bupropion SR for smoking cessation: data from clinical trials and five years of postmarketing experience. AB - Bupropion SR was introduced for smoking cessation in the US in 1997. This review assesses the efficacy and safety of bupropion SR for treatment of tobacco dependence based on data from clinical trials and five years of postmarketing experience. Through June 2001, there were approximately 32 million patient exposures to bupropion (9 million for smoking cessation) in clinical practice, and more than 8000 patients have been studied in clinical trials for tobacco dependence. In clinical trials, bupropion SR was more effective than placebo at improving initial and long-term abstinence rates and preventing relapse. Bupropion SR is generally well tolerated. The most common adverse event in clinical trials or clinical practice is insomnia, which can also be a symptom of nicotine withdrawal. The two main risks of treatment with bupropion SR are major motor seizure and hypersensitivity reaction. Clinical trials data suggest that the incidence of seizure is approximately 0.1%, and that of serious cases of hypersensitivity approximately 0.12%. Benefit-risk assessment, assuming a 30% one year quit rate demonstrates that for every 10,000 smokers treated with bupropion SR, 19 lives are saved and 86 cases of smoking-attributed morbidity are averted in a five-year period while the risk of experiencing one of the two potentially serious adverse events during treatment is 0.22%. These data further establish both the efficacy and safety of bupropion SR and its use in preventing the adverse health effects of chronic tobacco use. PMID- 12723729 TI - Anakinra: the first interleukin-1 inhibitor in the treatment of rheumatoid arthritis. AB - Rheumatoid arthritis is an immunologically mediated inflammation of joints of unknown aetiology and often leads to disability. This inflammatory process may also involve extra-articular connective tissue. New therapeutic approaches have been made by inhibition of proinflammatory cytokines. Interleukin-1 (IL-1) is regarded as one of the most important mediators in the development of synovialitis. In this article, anakinra (Kineret), the first direct antagonist to IL-1, is discussed, in particular the efficacy and safety data from clinical trials. More than 10,000 patients have been treated with anakinra with significant improvement of inflammation and pain; the rate of radiologically visible progressive joint damage was significantly reduced. Among the adverse events, injection site reactions were most frequent, followed by a mild increase in infections. No activation of tuberculosis, as in tumour necrosis factor-alpha antagonist administration, has so far been reported. PMID- 12723730 TI - Acute disseminated encephalomyelitis: a riddle wrapped in a mystery inside an enigma. AB - The difficulties in establishing a diagnosis of acute disseminated encephalomyelitis (ADEM) are illustrated in this case report, which attempts to raise awareness of the condition among general physicians, since they are the practitioners most likely to see such acute neurological presentations. Clinical and investigative features which raise the index of suspicion for the diagnosis of ADEM are outlined. PMID- 12723731 TI - A prospective evaluation of the management of acute pyelonephritis in adults referred to urologists. AB - In order to assess whether it is appropriate and clinically efficient to admit adults with 'clinically diagnosed' acute pyelonephritis (APN) under urologists, as is current practice in many NHS hospitals, a prospective study was undertaken over nine months in an NHS teaching hospital. Thirty-nine patients with clinical APN were admitted to the urology unit; all were pyrexial and 30 (77%) had typical features of rigor, flank pain and irritative lower urinary tract symptoms. Twenty one (54%) had positive urine cultures, 31 (79%) had parenteral antibiotics, while another three (7%) had oral agents initially. The remaining five (14%) were continued on agents initiated by their GPs before admission. Thirty-three (85%) had imaging procedures with eight significant anomalies being noted. Urgent invasive intervention was required in only four (10%) patients; length of stay varied from one to 25 days. Uncomplicated moderate to severe APN in adults may be treated safely without the need for admission to the urology unit, either in the outpatient setting or on an acute admissions observation ward. Complicated cases requiring intervention can be transferred to the urologist once recommended investigations have been undertaken. This care pathway may help to reduce cancellations of elective urological cases and is likely to be more cost effective for the NHS by reducing unnecessary admissions. PMID- 12723732 TI - Sarcoidosis emerging after adrenalectomy for Cushing's syndrome. AB - Cushing's syndrome is characterised by circulating steroid excess; the mainstay of treatment of systemic sarcoidosis is steroid therapy. We describe a case of sarcoidosis emerging after treatment for Cushing's syndrome. PMID- 12723733 TI - Acute rhabdomyolysis due to prolonged exposure to the cold. AB - Rhabdomyolysis represents an acute skeletal muscle injury and may be accompanied by acute renal failure due to myoglobinuria. This disorder has many potential causes. We describe the case of a 75-year-old man who developed rhabdomyolysis after prolonged exposure to cold but without associated hypothermia. This unusual case highlights that an appropriate investigation for rhabdomyolysis such as serum CK measurement should be performed in patients presenting with a history of prolonged exposure to cold, even in the absence of an obvious precipitating factor. PMID- 12723734 TI - Spontaneous rupture of a normal spleen. AB - A 45-year-old woman was admitted with a gangrenous toe. She had a pancreatico renal transplant for end-stage renal disease due to type I insulin dependent diabetes three years previously and had been taking immunosuppressive agents. An epidural catheter was placed to relieve the rest pain. Next day she was found to be hypotensive and tachycardic along with other features of shock. She did not have any clinical symptoms and signs initially. Urgent blood investigations were normal except for Hb of 5.0 g/dl; an ultrasound scan showed free fluid in the peritoneal cavity. Emergency laparotomy after resuscitation confirmed a massive haemoperitoneum secondary to a ruptured spleen. Histology showed loss of areas of the capsule and other areas showing subcapsular haemorrhage with no underlying pathology in the spleen. PMID- 12723735 TI - Bilateral renal artery stenosis after abdominal radiotherapy for Hodgkin's disease. AB - A 49-year-old woman was admitted with fatigue, dyspnoea, pretibial oedema and decreased daily urination. Seven years ago she was treated with doxorubicin, bleomycin, vinblastine and dacarbazine, alternating with mechlorethamine, vincristine, procarbazine and prednisone and 80 Gy abdominal radiotherapy for Hodgkin's disease. Two years later, malignant hypertension was diagnosed. Angiotensin-2 antagonist and beta-blocker treatment was given. After increased serum creatinine levels were determined, renal angiography was performed and total obstruction in the left renal artery and near total obstruction in the right side was observed. She was admitted to our clinic with oliguria, and acute renal failure was diagnosed. Balloon angioplasty and stent implantation was performed to the right renal artery. After a polyuric period, serum creatinine reduced to near normal levels. Angiotensin-2 antagonist treatment worsened the course in this patient. Patients with resistant hypertension occurring years after abdominal radiotherapy should be evaluated for renal artery stenosis. PMID- 12723736 TI - Combination fibrate-statin therapy for the treatment of severe hypertriglyceridaemia in renal disease. AB - This case report describes the treatment of a patient with end-stage renal disease on peritoneal dialysis who presented with triglycerides of 35 mmol/l, cholesterol of 14 mmol/l and visible chylomicronaemia. She was only partially responsive to statin and fibrate monotherapy, so was treated with fibrate-statin combination therapy. Daily fibrate therapy resulted in a myositis, but once weekly fibrate therapy with nocturnal daily statin therapy resulted in a reduction in total cholesterol of 49%, triglycerides of 82% and raised HDL by 52%, enabling her to achieve a target LDL of 3 mmol/l and HDL of 1 mmol/l. Combination therapy with low frequency (weekly) fibrate administration and daily statin treatment may provide a means to control the severe hypertriglyceridaemia of patients with renal disease. PMID- 12723737 TI - Does C-reactive protein represent an oxidative stress marker in cardiovascular disease? PMID- 12723738 TI - The mechanism of action of anti-inflammatory drugs. PMID- 12723739 TI - Discovery, mechanisms of action and safety of ibuprofen. AB - Ibuprofen was the product of a long research programme during the 1950s and 1960s to develop a 'super aspirin' for the treatment of rheumatoid arthritis which was as effective as current alternatives but safer. Selected for development in 1964 after several promising compounds had proved disappointing at the clinical stage, ibuprofen was found to have a short elimination half-life and exceptional gastrointestinal tolerability. Ibuprofen was introduced in the United Kingdom in 1966 and in the United States in 1974, and was the first non-steroidal anti inflammatory drug (NSAID) licensed for over-the-counter use in the UK in 1983 and in the US the following year. Ibuprofen is a non-cyclo-oxygenase selective NSAID but recent evidence suggests additional anti-inflammatory properties are due to modulation of leucocyte activity, reduced cytokine production, inhibition of free radicals and signalling transduction. Ibuprofen may also exert a central analgesic action in the dorsal horn. Future roles for ibuprofen may include protection against certain cancers and Alzheimer's disease. PMID- 12723741 TI - Review of the analgesic efficacy of ibuprofen. AB - There is a clear relationship between single doses of ibuprofen over the range 50 400 mg and the peak analgesic effect and the duration of analgesia. The smallest clinically useful dose of ibuprofen is 200 mg. Ibuprofen 400 mg has been shown to be as effective as aspirin 600 or 900 mg/day in models of moderate pain but superior to aspirin or paracetamol in more sensitive models such as dental pain. The duration of action of ibuprofen 400 mg is at least 6 hours compared with 4-6 hours for ibuprofen 200 mg or paracetamol. In patients undergoing oral surgery, ibuprofen 200 mg was broadly comparable with naproxen 220 mg and ibuprofen 400 mg comparable with ketoprofen 25 mg. The combination of ibuprofen and hydrocodone is more effective than either drug alone in patients undergoing abdominal and gynaecological surgery. The absorption of ibuprofen acid is influenced by formulation, and certain salts of ibuprofen (lysine, arginine, potassium) and solubilised formulations have an enhanced onset of activity. These differences are clinically important, offering a shorter time to onset of relief of tension headache compared with paracetamol. PMID- 12723740 TI - A general overview of the use of ibuprofen in paediatrics. AB - Ibuprofen is prescribed for children for the treatment of acute pain and fever, and for juvenile idiopathic arthritis. The pharmacokinetic characteristics of ibuprofen in children are similar to those in adults and the relationship between dose and response is linear over the range 5-10 mg/kg. Clinical trials of ibuprofen have shown the effective dose range to be 7.5-10 mg/kg. The maximum reduction in temperature occurs 3-4 hours after administration. In comparative clinical trials, ibuprofen has been shown to be equally as effective as or more effective than paracetamol as an analgesic and antipyretic and to have a longer duration of action; it is also as effective as aspirin. The adverse effects of ibuprofen are similar to those of other non-steroidal anti-inflammatory drugs but clinical experience suggests that ibuprofen is better tolerated by children than adults and it is safer in overdose than paracetamol and aspirin. PMID- 12723743 TI - The use of ibuprofen in neonates in the treatment of patent ductus arteriosus. AB - Indomethacin has long been used to treat patent ductus arteriosus but it is associated with a relatively high risk of adverse effects; recent evidence suggests that ibuprofen is effective and may be safer. In a randomised trial to compare the efficacy and safety of ibuprofen and indomethacin in the treatment of patent ductus arteriosus, 144 infants received three doses of ibuprofen lysine (10, 5 and 5 mg/kg) at 24-hour intervals or indomethacin 0.2 mg/kg at 12-hour intervals. Ductal closure occurred in 70% of children treated with ibuprofen and 66% of those given indomethacin on the first treatment (P = 0.41). Nineteen children underwent surgical ligation, equally distributed between the treatment groups (P = 0.81). Urine production was significantly greater than in children given indomethacin from day 3 to day 7 and the serum creatinine concentration was significantly lower from day 7. Ductal closure was associated with higher serum concentrations of ibuprofen and a concentration of 10-12 mg/l appears to be the minimum level for efficacy. In a randomised, placebo-controlled, double-blind trial of prophylaxis with ibuprofen, the rate of patent ductus arteriosus associated with ibuprofen was 19% compared with 42% with placebo. Urine output was comparable in the two groups except on day 1, when it was significantly lower among infants given ibuprofen. Ibuprofen is therefore as effective as indomethacin in the treatment of patent ductus arteriosus, and effective as prophylaxis, in premature infants. PMID- 12723742 TI - Relative efficacy of selective COX-2 inhibitors compared with over-the-counter ibuprofen. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) suppress the activity of both isoforms of cyclo-oxygenase (COX). Inhibition of COX-1, the constitutive isoform, is primarily responsible for the adverse gastrointestinal effects of the NSAIDs whereas inhibition of COX-2, the inducible isoform, accounts for their therapeutic effects. COX-2 inhibitors such as celecoxib and rofecoxib appear to be as effective as non-selective NSAIDs in the treatment of chronic inflammatory disease but their analgesic efficacy and their safety at the higher doses required for analgesia are less certain. There is consistent evidence that COX-1 plays a major role in the early pain response following injury and that analgesia is increased when both COX-1 and COX-2 are inhibited simultaneously. Early postoperative nociception may cause hyperalgesia at a later time by a process of central plasticity. In an experimental model of pain, ibuprofen promptly suppresses prostaglandin E2 concentrations whereas celecoxib has no discernible effect until 90-120 minutes postoperatively, when COX-2 activity is induced. Both drugs significantly reduce pain compared with placebo but celecoxib appears to have a slower onset of action. The analgesic effect of ibuprofen is well characterised for acute pain and short-term treatment is well tolerated. PMID- 12723745 TI - Risk factors associated with the development of gastroduodenal ulcers due to the use of NSAIDs. AB - The risk of gastrointestinal mucosal injury with non-steroidal anti-inflammatory drugs (NSAIDs) is dose-dependent. Epidemiological studies have clearly demonstrated a rank order of risk of ulcer complications for commonly used NSAIDs, with ibuprofen consistently associated with the lowest risk and piroxicam with the highest. Antacids, H2 receptor antagonists and misoprostol all have drawbacks as prophylaxis. Of the cyclo-oxygenase (COX)-2 selective NSAIDs, rofecoxib is associated with a lower risk of gastrointestinal toxicity but there is uncertainty about the long-term risk associated with celecoxib. Rofecoxib has been associated with a significantly higher incidence of myocardial infarction than naproxen that may counteract the benefit of greater gastrointestinal safety. At over-the-counter doses, the short duration of use and the low dose reduce the risk of a serious adverse event compared with chronic use at prescribed doses. Intermittent therapy with low-dose NSAIDs has proved extremely safe and it has not been determined whether COX-2 selective agents offer a safety advantage compared with such treatment. PMID- 12723744 TI - Forty years of ibuprofen use. AB - Low-dose ibuprofen is as effective as aspirin and paracetamol for the indications normally treated with over-the-counter (OTC) medications and is associated with the lowest risk of gastrointestinal toxicity of any non-steroidal anti inflammatory drug. By contrast, even low-dose aspirin is associated with an appreciable risk of gastrointestinal toxicity. Paracetamol is well tolerated and effective in treating mild to moderate pain but there is growing concern about a possible risk of gastrointestinal toxicity and a possible link with asthma in children. The PAIN (Paracetamol, Aspirin, Ibuprofen New tolerability) study was a blinded randomised comparison of the tolerability of OTC analgesics in the treatment of common types of acute pain encountered in the community. A total of 8,677 adults were randomised to treatment with ibuprofen 1200 mg/day, paracetamol 3 g/day or aspirin 3 g/day for 1-7 days. The most common indications for treatment were musculoskeletal conditions (31-33%), colds or flu (19-20%), backache (15-17%), sore throat (11-12%) and headache (10-11%). Significant adverse events were more common with aspirin (10.1%) than ibuprofen (7.0%) (P<0.001) or paracetamol (7.8%). Significant gastrointestinal events were less frequent with ibuprofen (4.0%) than with aspirin (7.1%, P<0.001) or paracetamol (5.3%) (P=0.025). For every 100 patients treated, five more will experience significant adverse events if they are taking aspirin rather than ibuprofen, and four more than if they were taking paracetamol. PMID- 12723747 TI - Epidemiology overview of gastrointestinal and renal toxicity of NSAIDs. AB - This review updates previous systematic reviews to explore the overall levels of risk of serious upper gastrointestinal complications of treatment with non steroidal anti-inflammatory drugs (NSAIDs) and to investigate the importance of dose in explaining variations in risk. Thirty-six eligible case control studies were published between 1985 and 2000 and involved 19,648 cases and 105,373 controls. Eight eligible cohort studies included around 400,000 exposed subjects and 1 million non-exposed controls. The pooled unadjusted odds ratio from case control studies (compared with non-use of NSAIDs) for serious gastrointestinal complications is 4.06 (CI95% 3.47, 4.75); the pooled odds ratio from cohort studies is 2.29 (CI95% 1.50, 3.51). Unadjusted odds ratios range from 1.81 for ibuprofen to 7.46 for piroxicam. These data require careful interpretation because the statistical power of analyses of individual NSAIDs is lower than that for the class as a whole. There is a rank order of risk for different NSAIDs at low doses but at higher doses the odds ratios tend to converge. Ibuprofen is associated with a lower risk of serious gastrointestinal complications than other NSAIDs; this advantage is probably lost at higher doses (>1800 mg/day). PMID- 12723746 TI - Population impact of strategies designed to reduce peptic ulcer risks associated with NSAID use. AB - The risk of ulcer complications rises steeply with dose for aspirin and other non steroidal anti-inflammatory drugs (NSAIDs) but estimates of the overall incidence of bleeding ulcer are unreliable. Drug utilisation data, epidemiological data on the frequency of bleeding ulcer and death, and the relative risks associated with different NSAIDs, indicate that the number of cases of bleeding ulcer attributable to NSAIDs in the United Kingdom is approximately 2,400. Substitution of ibuprofen at a dose of 2.4 g/day for all other NSAIDs would reduce the number of events attributable to NSAIDs from 2,431 to 695 annually. At a dose of 1200 mg/day, substituting ibuprofen or another safe NSAID would be likely to reduce events to zero. Similarly, substitution of ibuprofen 2.4 g/day for all other NSAIDs would reduce attributable ulcer mortality to 80. The total number of excess cases attributable to aspirin is 753 annually. If prophylactic aspirin was prescribed solely at a dose of 75 mg/day, the number of cases would fall to 445 annually and the number of related deaths from 87 to 51 annually. NSAIDs and aspirin account for approximately one-third and previous ulcer for about one fifth of the overall risk of bleeding ulcer and its complications. PMID- 12723748 TI - The safety of ibuprofen suspension in children. AB - This paper describes two studies in children with fever in which the safety of ibuprofen was compared with that of paracetamol. The Boston University Fever Study aimed to assess the risk of rare but serious adverse events in febrile children. There were 795 admissions among 84,192 children during the study. There were no significant differences between the drugs in the risk of admission or the risk of secondary endpoints (admissions for asthma or cellulitis, or physician visits for abdominal pain or dyspepsia) and no evidence of clinically significant impairment of renal function. However, ibuprofen was associated with a significantly lower risk of physician visits for asthma: the incidence associated with ibuprofen was 3.0% (CI95% 2.1, 4.1) compared with 5.1% (CI95% 3.5, 7.1) for paracetamol (P = 0.02). The second study was a case control study to investigate a possible association between antipyretic medication, varicella infection and necrotising fasciitis. We identified 52 children aged under 19 years who were admitted to hospital with varicella and Group A streptococcal infection and 172 matched controls with uncomplicated varicella. The risk of invasive Group A streptococcal infection was associated with demographic and environmental factors and persistent high fever. There was no association with the use of ibuprofen or paracetamol alone, but the use of both agents was significantly associated with streptococcal infection. These studies demonstrate that children with fever tolerate treatment with ibuprofen as well as treatment with paracetamol. Neither agent is associated with an increased risk of necrotising soft tissue infections. PMID- 12723749 TI - Ibuprofen overdose. AB - Enquiries about the management of patients possibly suffering from ibuprofen overdose account for over 5% of the total enquiry workload of the London Centre of the United Kingdom National Poisons Information Service. Unlike overdose with aspirin and paracetamol, no additional pathophysiological findings have been reported in ibuprofen overdose and all the demonstrated toxic effects relate to its known pharmacological actions and the effects of accumulation of the two acidic metabolites, 2-hydroxyibuprofen and 2-carboxylibuprofen. The most striking finding in reported cases is that the great majority of patients suffer no or only mild symptoms. In one series of 1,033 enquiries involving ingestion of ibuprofen alone, 705 (65%) patients were asymptomatic; 199 (18%) experienced mild symptoms; and 23 (2%) experienced moderate symptoms. We are aware of only seven case reports of fatal overdose with ibuprofen and in each case there are complicating factors related to other drugs and/or other disease processes. The management of ibuprofen overdose is generally straightforward and can be related to the dose ingested. Initial findings suggest even less evidence for toxicity associated with modified release formulations than with the conventional tablets. There is at present no reason to be concerned that co-ingestion of ethanol increases the risk of toxicity from ibuprofen overdose. Ibuprofen overdose is common but serious toxic effects are unusual and guidelines for treatment are straightforward. PMID- 12723750 TI - Polysaccharide peptide mediates apoptosis by up-regulating p21 gene and down regulating cyclin D1 gene. AB - The use of herbal medicine is a common practice among Chinese women with breast cancer. Yunzhi (Voriolus versicolor), a substance that is regarded as a biological response modifier, is frequently used. The aim of the present study is to evaluate the anti-proliferative action of, Yunzhi, polysaccharide peptide (PSP), on breast cancer cells. Breast cancer cells (MDA-MB-231) were cultured with and without PSP for 7 days. Cell growth at 24, 72, 120 and 168 hours was measured by Cell Proliferation Reagent (WST-1). Cells treated with PSP were found to have a significant reduction in cell proliferation as compared to controls after 72 hours of incubation. This lasted for 168 hours. When the effect of PSP on apoptosis was studied by the TdT-mediated X-dUTP nick end-labeling (TUNEL) assay, we found that PSP had a significant effect upon apoptosis from 24 hours onward. Immunostaining showed that PSP increased p21 expression and decreased cyclin D1 expression. In conclusion, PSP is effective in inhibiting cell proliferation through apoptosis. The mechanism for the apoptosis may be through up-regulation of p21 and down-regulation of cyclin D1. PMID- 12723751 TI - Effect of polysaccharide Krestin on the up-regulation of macrophage colony stimulating factor gene expression in protecting mouse peritoneal macrophages from oxidative injury. AB - Oxidative injury caused by oxidatively modified low density lipoprotein (Ox-LDL) plays an important role in the transformation of macrophages into foam cells and atherogenesis. Treatments to protect macrophages from oxidative injury will be effective in treating atherosclerosis. A macrophage-specific growth factor, macrophage colony-stimulating factor (M-CSF), was reported to be able to prevent the progression of atherosclerosis in Watanabe heritable hypercholesterolemic (WHHL) rabbits. A protein-bound polysaccharide, polysaccharide Krestin (PSK), was also proven to have effects in preventing atherosclerosis in our previous work. We proposed that, both M-CSF and PSK could protect macrophages from oxidative injury, and the effects of PSK were associated with its capability of inducing M CSF expression. In our present results, M-CSF could alleviate the Ox-LDL- or tert butyl hydroperoxide (tbOOH)-induced injury to mouse peritoneal macrophages, and PSK exhibited some similar effects. PSK treatment could induce M-CSF gene expression and secretion in mouse peritoneal macrophages. Furthermore actinomycin D and cycloheximide could attenuate that induction. We concluded that, maybe PSK exerted its effects on macrophages partly through the transcriptional induction of M-CSF in the cells. PMID- 12723752 TI - Scavenging of reactive oxygen species and inhibition of the oxidation of low density lipoprotein by the aqueous extraction of Anoectochilus formosanus. AB - The ability of Anoectochilus formosanus extract (AFE) to react with relevant biological oxidants was evaluated in this study. In addition, its effect on oxidation of low density lipoprotein (LDL) was investigated in vitro and in vivo. AFE could scavenge reactive oxygen species, such as superoxide anion and hydroxyl radical. The study of human LDL oxidation showed that AFE delayed oxidation in a concentration-dependent manner. In vivo studies also showed that oral administration of AFE delayed the oxidation of LDL from hyperlipidemic hamsters. The ability of AFE to scavenge free radicals suggests that it may be a promising anti-atherogenic agent. PMID- 12723753 TI - Antileukemic activity of selected natural products in Taiwan. AB - The aim of the present study was to evaluate the antileukemic activity of six chemical classes of pure compounds present in commonly used medicinal plants in Taiwan--such as the genus Plantago. Studies were conducted on a series of human leukemia and lymphoma cell lines. Results showed that water soluble compounds (aucubin, caffeic acid, chlorogenic acid, ferulic acid, p-coumaric acid and vanillic acid) exhibited a weak antileukemic activity (IC50: 26-56 microg/ml, SI: 2-11). On the other hand, water insoluble compounds such as triterpenoids (oleanolic acid and ursolic acid), monoterpene (linalool) and flavonoid (luteolin) possessed strong activity against human leukemia and lymphoma cell lines. Among them, linalool showed the strongest activity against histiocytic lymphoma cells U937 (IC50: 3.51 microg/ml, SI: 592.6) and Burkitt lymphoma cells P3HR1 (IC50: 4.21 microg/ml, SI: 494.1). Ursolic acid was effective against P3HR1 cells (IC50: 2.5 microg/ml, SI: 262.6) and chronic myelogenous leukemia cells K562 (IC50: 17.79 microg/ml, SI: 36.91), whereas oleanolic acid inhibited the growth of P3HR1 cells (IC50: 26.74 microg/ml, SI: 11.37). Luteolin exhibited effective activity against K562 cells (IC50 18.96 microg/ml, SI: 5.14) and P3HR1 cells (IC50: 18.99 microg/ml, SI: 5.13). We conclude that terpenes and flavonoid in commonly used medicinal plants possess strong activity against lymphoma and leukemia cells, especially human lymphoma cells, suggesting the potential use of these compounds for treatment of lymphoma. PMID- 12723755 TI - Evaluation of antinociceptive, anti-inflammatory and antipyretic effects of Strobilanthes cusia leaf extract in male mice and rats. AB - The leaf of Strobilanthes cusia (Acanthaceae), popularly known as Da-Ching-Yeh, has been commonly used in traditional Chinese medicine. It is used for influenza, epidemic cerebrospinal meningitis, encephalitis B, viral pneumonia and mumps. It is also used to treat sore throat, aphthae and inflammatory diseases with redness of skin, etc. In this study, we evaluated the antinociceptive, anti-inflammatory and antipyretic effects of methanol extract of Strobilanthes cusia leaf. The results showed that the extract significantly inhibited the writhing responses of mice and decreased the licking time on both the early and late phases of the formalin test in a dose-dependent manner. It also reduced the paw edema induced by carrageenan in rats. In addition, it potently attenuated pyrexia induced by lipopolysaccharide. PMID- 12723754 TI - Anxiolytic effect of ting-chih-wan in mouse behavior models of anxiety. AB - The anxiolytic effect of the alcoholic extract of ting-chih-wan (TCWa) was studied using the black and white test (BWT) and the elevated plus-maze (EPM). We further demonstrated the anxiolytic mechanism of TCWa by combining with diazepam (DIZ), serotonin (5-HT) agonists or antagonists, and measuring the levels of monoamines and its metabolites in the brain stem and cortex. In the BWT, TCWa (0.1-1.0 g/kg, p.o.) increased the time spent in the white chamber and total change between the two chambers, and decreased the time spent in the black chamber. TCWa (0.1-0.5 g/kg, p.o.) increased the arm entries and the time spent on the open arms, and decreased the arm entries and the time spent on the closed arms in the EPM. On the other hand, TCWa (1.0 g/kg, p.o.) decreased horizontal activity and prolonged pentobarbital-induced sleeping times. TCWa (0.1, 0.5 g/kg) decreased the levels of norepinephrine (NE), dopamine (DA), 5-HT and 5-hydroxy indoleacetic acid (5-HIAA) and increased the levels of vanillylmandelic acid (VMA) and homovanillic acid (HVA) in the brain stem. TCWa (0.1 and 0.5 g/kg) decreased the levels of NE, DA and increased the levels of VMA and HVA in the cortex. TCWa also attenuated the anxiogenic effect of 5-hydroxytryptophan (5-HTP) and enhanced the anxiolytic effect of 9p-chlorophenylalanine (PCPA), buspirone (BUS) and ritanserin (RIT) in the EPM. From these results, TCWa at 0.1 and 0.5 g/kg possessed an anxiolytic effect T heanxiolytic mechanisms of TCWa might be due to decreased catecholaminergic activity caused by the increase in the turnover rate of catecholamines in the brain and decreased concentrations of 5-HT in the brain stemvia activating somatodendritic 5-HT1A autoreceptors and inhibiting postsynaptic 5-HT receptors. PMID- 12723756 TI - Long-term administration of Polygonum multiflorum Thunb. reduces cerebral ischemia-induced infarct volume in gerbils. AB - Focal cerebral ischemia was produced by an occlusion of the middle cerebral artery for 1, 3 and 24 hours in gerbils. Infarct volumes were determined by 2,3,5 triphenyltetrazolium chloride (TTC) transcardiac perfusion 24 hours after cerebral ischemia. Significant and consistent infarct sizes were produced in gerbils subjected to 24-hour occlusion of the middle cerebral arterey when compared to the 1 and 3-hour occlusion group. Longterm pretreatment of the 50% ethanol extract of Polygonum multiflorum Thunb. for 2 weeks significantly reduced the infarct volume by 50% as compared to that of the 24-hour occlusion group. The results revealed that long term pretreatment of Polygonum multiflorum Thunb. may protect the brain against focal cerebral ischemia. PMID- 12723757 TI - Choto-san prevents occurrence of stroke and prolongs life span in stroke-prone spontaneously hypertensive rats. AB - The effects of long-term oral administration of choto-san (diao-teng-san in Chinese) extract on the occurrence of stroke and life span were investigated in stroke-prone spontaneously hypertensive rats (SHR-SPs). Twenty-four rats were ramdomized into three groups. From 8 weeks of age, 0.1% and 0.3% choto-san groups were given water containing 0.1% (150 mg/kg/day) and 0.3% (450 mg/kg/day) choto san extract, respectively. A control group was given only water. The mean survival times of the control group, 0.1% and 0.3% choto-san groups were 122.1, 159.8 and 176.8 days, respectively. The percent survivals of both the 0.1% and 0.3% choto-san groups were significantly enhanced compared to the control (Kaplan Meier analysis followed by log-rank test; 0.1% choto-san: p < 0.05; 0.3% choto san: p < 0.05). Furthermore, the cumulative percent occurrence of neurological and behavioral signs accompying stroke in the 0.3% choto-san group was significantly inhibited compared to the control (p < 0.05). These results suggested that choto-san prevents the occurrence of stroke and prolongs the life span of SHR-SPs. PMID- 12723758 TI - Effects of a herbal extract on the bone density, strength and markers of bone turnover of mature ovariectomized rats. AB - For many decades, the Chinese have been using herbal medications to treat bone diseases. To examine effects of an extract of ten medicinal herbs on estrogen deficiency bone loss, ten-month-old female rats were randomly divided into three groups: ovariectomized (OVX), OVX treated with herbs (OVX-M) 4 ml/day by gavage, and OVX treated with estrogen (OVX-E) 10 mg subcutaneously (s.c.) twice per week. The bone mineral density (BMD) of the left femur (fBMD), spine (sBMD) and global body (gBMD) were measured at baseline and at 4, 8 and 12 weeks using a Hologic QDR 2000 dual-energy X-ray densitometer. Tibial strength was tested using the Instron Model 5566 electro-mechanical testing machine. The urinary pyridinoline creatinine ratio (Pyd/Cr), deoxypyridinoline creatinine ratio (Dpd/Cr), plasma alkaline phosphatase (ALP), calcium (CA), phosphorus (P) and albumin (ALB) were also determined. Uterine weight was determined at 12 weeks. The results showed that percent changes of fBMD in the OVX (n = 9), OVX-E (n = 8) and OVX-M (n = 8) rats at the 12-week time point were -11.8 +/- 4.6(c), 1.8 +/- 3.1(a), -7.6 +/- 1.9(abc) (p < 0.05-0.001, a: vs. OVX, b: vs. OVX-E, c: vs. baseline); sBMD were 10.7 +/- 4.6(c), -0.3 +/- 5.5(a), -5.9 +/- 3.5(abc); and gBMD were -4.8 +/- 2.3(c), 0.1 +/- 2.4(a), -2.7 +/- 2.6(abc), respectively. Further, the tibia maximum breaking stress and flexural modulus of elasticity in OVX-M rats (295 +/- 33(a), 18,194 +/- 3,264(a)) were significantly higher (p < 0.005-0.001) than that in OVX rats (189 +/- 83, 10,309 +/- 4,930), and similar to OVX-E rats (298 +/- 35(a), 18,766 +/- 2,620(a)). Additionally, the herbal extract reduced the urinary Pyd/Cr, Dpd/Cr and plasma ALP increment followed OVX and was not associated with a rise in uterine weight. In conclusion, the herbal extract demonstrated a therapeutic effect to inhibit bone resorption and to reduce estrogen-dependent bone loss without uterine stimulation. It may have potential as a new approach in treating and preventing postmenopausal osteoporosis (PMOP). PMID- 12723759 TI - Effects of goshajinkigan on corneal sensitivity, superficial punctate keratopathy and tear secretion in patients with insulin-dependent diabetes mellitus. AB - Goshajinkigan (niu-che-shen-qi-wan in Chinese), a traditional herbal medicine, has been used in Japan to treat clinical symptoms of diabetic neuropathy. A double-masked study was performed to evaluate its effects on corneal sensitivity, superficial punctate keratopathy and tear production in patients with insulin dependent diabetes mellitus. Fifty diabetic patients were randomized into two groups: Group A, in which 25 patients received goshajinkigan orally, 7.5 g/day for 3 months; Group B, in which 25 patients were orally administered placebo, 6.0 g/day for 3 months; and in Group C, 25 non-diabetic subjects were orally administered goshajinkigan, 7.5 g/day for 3 months. Corneal sensitivity was measured with an aesthesiometer. The area of superficial punctate keratopathy was expressed as a fluorescein staining score. Reflex tearing was determined with a Schirmer test without anesthesia goshajinkigan was analyzed by high-performance liquid chromatography. Corneal thresholds after treatment with goshajinkigan (2.03 g/mm2) in Group A were significantly lower than those before treatment (2.47 g/mm2). Those in Groups B and C did not change after treatment. Fluorescein staining scores after administration of Goshajinkigan (0.64) in Group A were significantly lower than those before treatment (1.32). Those in Groups B and C did not change after treatment. Schirmer test results after goshajinkigan administration (11.0 mm/5 min) in Group A were significantly higher than those before treatment (9.3 mm/5 min). Those in Groups B and C did not change after treatment. Hemoglobin A1c levels in Groups A, B,and C did not change after treatment. Several components in goshajinkigan were found on high performance liquid chromatography. In conclusion, goshajinkigan improved ocular surface disorders in patients with insulin-dependent diabetes mellitus. PMID- 12723760 TI - Hepatoprotective activity of reduohanxiao-tang (yuldahanso-tang) is related to the inhibition of beta-glucuronidase. AB - Beta-glucuronidase-inhibitory and hepatoprotective effects of Reduohanxiao-tang (Yuldahanso-tang), which has been used for liver diseases and stroke, on carbon tetrachloride (CCl4)-induced hepatotoxicity of rats were investigated. Reduohanxiao-tang potently inhibited beta-glucuronidases. Serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactic acid dehydrogenase (LDH) levels of the CCl4 group orally treated with Reduohanxiao tang (100 mg/kg) were lowered to 54%, 71.5% and 66.1% of the CCl4-treated control group, respectively. Among the ingredients of the Reduohanxiao-tang, the rhizomes of Pueraria thunbergiana and Scutellaria baicalensis potently inhibited beta glucuronidases and protected against CCl4-induced liver injury. Orally administered puerarin, which is a main component of Pueraria thunbergiana, showed potent hepatoprotective activity, but did not inhibit beta-glucuronidase. However, daidzein, which is produced from puerarin by human intestinal bacteria, potently inhibited beta-glucuronidase. These results suggest that beta glucuronidase inhibition by herbal medicines may protect against CCl4-induced liver injury. PMID- 12723761 TI - Traditional Chinese medicine prevents inflammation in CCl4-related liver injury in mice. AB - Alternative medicines are being increasingly used and investigated in the management of a variety of disorders. Hepatitis is a common indication for the use of alternative therapies but evidence for the efficacy of many compounds is lacking. We have utilized a well-defined model of liver injury to study the efficacy of three herbal products designed to assist in the management of liver disease. Mice were exposed to carbon tetrachloride (CCL4) given intragastrically after they had been pretreated for five days with either saline or one of four doses of silymarin extract or CH100 (a Chinese herbal medicine comprising of 19 herbs) or one of two doses of CH101 (a Chinese herbal preparation designed to reduce fibrosis). Animals were sacrificed 24 hours after receiving CCL4. Liver enzymes and hepatic histology formed the basis for evaluating efficacy of the treatments. Each of the alternative medicines reduced the alanine amino transferase (ALT) elevation demonstrated after CCL4 injection. The high dose CH100 regimen was most effective in protecting against injury and this was confirmed with hepatic histology. Other doses of CH100, CH101 and silymarin were not shown to provide protection against the histological damage. In conclusion, Silymarin, CH100 and CH101 are able to reduce ALT elevation in animals exposed to CCL4. High dose CH100 provides protection from hepatocyte necrosis in this model. The data add to our understanding of the capacity some herbal medicines have to modify the reaction of the liver to a variety of insults and suggest the value of studying these agents further in human liver diseases. PMID- 12723762 TI - Effect of dai-kenchu-to on obstructive bowel disease in children. AB - The aim of this study was to evaluate the effects of a traditional Chinese herbal medicine, dai-kenchu-to (DKT), on obstructive bowel diseases in children. We have treated 46 pediatric patients with various obstructive bowel diseases with DKT: six patients with postoperative ileus, 12 with large abdominal surgery (including three neonates), one with ano-rectal anomaly, three with Hirschsprung's disease, two with functional bowel obstructions, one with SMA syndrome, and 21 patients with chronic constipation. DKT (0.1-0.15 g/kg) was mixed with 5-10 ml of warm water, and was given orally two to three times a day. DKT was effective for 39 patients (85%) and their clinical symptoms improved. DKT was ineffective in seven patients: two with postoperative ileus, two with Hirschsprung's disease, and three with chronic constipation. DKT had mild but significant effects for various obstructive bowel diseases in children, while no side effects were encountered. Our current strategy for pediatric patients with obstructive bowel disease is to use DKT first and then test its efficacy. If DKT is effective, the regimen is continued. However, in cases where DKT is not effective, we will consider laparotomy or will further investigate the illness. PMID- 12723763 TI - Reexamination of the relation between menstrual cycle and kampo diagnosis, yin yang. AB - Several Chinese herbal medicine textbooks describe that a short menstrual cycle indicates "yang" status and a long cycle indicates "yin" status. However, we sometimes encounter yang patients with a long cycle or yin patients with a short cycle in daily practice. Therefore, we reviewed the relationship between yin-yang and length of menstrual cycle. A questionnaire addressing both menstrual cycle and several parameters for determining yin-yang was completed at the time of the patients' visit to the hospital. Patients were divided into yang or yin groups, and menstrual cycle was compared between the two groups. A short menstrual cycle was mostly observed in yin patients (27.42 +/- 1.73, 95% CI: 26.32-28.52, n = 12) and a long cycle was mostly observed in yang patients (31.63 +/- 3.96, 95% C.I: 29.51-33.74, n = 16). The difference in menstrual cycle between the two groups was significant (p = 0.002), and this result was entirely opposite to the traditional theory. This study identified a discrepancy between the traditional theory and the actual menstrual cycle findings in this small population. Current reexamination of the relationship between yin-yang and menstruation in a larger population is warranted. PMID- 12723764 TI - Qi-training enhances respiratory burst function and adhesive capacity of neutrophils in young adults: a preliminary study. AB - The main objective of this study is to examine the effect of Qi-training on the immune system, especially neutrophil bactericidal function. Nine healthy male subjects were studied for the effects of one bout of ChunDoSunBup (CDSB) Qi training on superoxide (O2- production and adhesion capacity of neutrophils at times immediately after (Post I) and 2 hours after the Qi-training (Post II). The Qi-training enhanced O2- production, reaction velocity and neutrophil adhesion capacity and there were significant differences at Post I compared to before Qi training (Pre). In addition, the number of white blood cells (WBC), monocytes and lymphocytes were changed significantly through Qi-training.Therefore, it seems that CDSB Qi-training may increase the resistance of trained individuals against common infection and inflammation. PMID- 12723765 TI - Effect of bee venom treatment in sows with oligogalactic syndrome postpartum. AB - The objective of this study was to determine the clinico-therapeutic effect of worker honeybee venom in sows with oligogalactic syndrome postpartum. Comparison between bee venom- and drug-treated groups was our main concern in the present study. Sows after parturition were assigned to bee venom- and drug-treated groups, respectively. In the bee venom-treated group, 22 sows were bee acupunctured once a day for 3 consecutive days. Honeybees (Apis mellifera L.) forbee acupuncture were about 15 days old after metamorphosis. Live bees were used to sting the acupoints known as yang-ming (ST-18, 1.5 cm lateral to the base of the last two pairs of teats) and jiao-chao (GV- , at the indentation between the base of tail and the anus). In the drug-treated group, 20 sows were intramuscularly injected with a standard dose of penicillin G (400,000 IU/head) once a day for 3 consecutive days. On post-treatment day 4, 85.0% of the drug treated group and 90.9% of the bee venom-treated group recovered from oligogalactic syndrome postpartum. The result suggested that apitherapy using worker honeybee is an effective treatment for sows with oligogalactic syndrome postpartum. PMID- 12723767 TI - Is smaller necessarily better? PMID- 12723766 TI - Analgesic effects of different acupoint stimulation frequencies in humans. AB - Transcutaneous electrical acupoint stimulation (TEAS) provides a convenient and standardized technique for pain treatment. The cold-pressor test is a simple and reliable model in humans for the induction of tonic pain. In this controlled study, the effects of TEAS on cold pressor-induced pain were evaluated in 22 healthy human subjects. Electrical stimulation at 4 Hz and 32 Hz was applied to He-Gu (LI 4) and Nui-Guan (P 6) acupoints for 15 minutes. Pain score ratings were evaluated at four time points from 30-170 seconds during the cold-pressor test. We observed an analgesic effect at both 4 Hz and 32 Hz of stimulation, and pain score rating reductions were statistically significant compared to control (p < 0.01). Our data support the efficacy of TEAS analgesia. However, there was no significant difference between pain scores at 4 Hz and 32 Hz stimulation. PMID- 12723768 TI - Posterior lateral corner reconstruction. PMID- 12723769 TI - Evaluation of osteoporosis in orthopedic practice: a review of current diagnostic modalities. AB - It is increasingly recognized that osteoporosis should be of utmost importance to all, because of the large size of the affected population and the staggering implications of osteoporotic fractures on morbidity, mortality, and medical costs. Osteoporosis is one of the most prevalent conditions associated with aging, and, as the demographics shift toward an older population, a dramatic increase in the number of osteoporotic fractures is expected in coming years. Fortunately, osteoporosis is a disease that can be easily diagnosed, owing to the growing armamentarium of diagnostic tests that should be familiar to orthopedic surgeons, radiologists, endocrinologists, nephrologists, rheumatologists, gynecologists, and generally all physicians with a special interest in bone metabolism. PMID- 12723770 TI - Patellar retinacular peel: an alternative to lateral retinacular release in total knee arthroplasty. AB - The need to perform a lateral retinacular release (LRR) during total knee arthroplasty (TKA) may be lessened or eliminated by performing a patellar retinacular peel (PRP). PRP involves peeling the extensor retinaculum off the dorsolateral border of the patella to the extent necessary to facilitate patellar eversion. Generally, a portion of the lateral patellar facet is removed to allow conformity with a round patellar prosthesis. The combination of retinacular peel and partial facetectomy decreases lateral retinacular tension, which reduces the tendency toward lateral subluxation. In the study reported here, a series of 50 TKAs that included a PRP in all cases was compared with an earlier series of 50 TKAs that included LRR for control of patellar tracking, as needed. The incidence of anterior knee pain was lower in the PRP group than in the LRR group, though this difference was not significant from a statistical perspective. The 2 groups were virtually identical on other comparison criteria. PRP seems to be a viable alternative to LRR. PMID- 12723771 TI - Mechanical therapy for loss of knee flexion. AB - We hypothesized that adding home mechanical therapy to traditional physical therapy by a physical therapist would significantly reduce the need for surgical management of loss of knee flexion after surgery or injury. From 1990 to 1999, we followed up on 34 patients who added home mechanical therapy after failure of physical therapy alone. Thirty-one (91.2%) of these patients regained functional flexion (defined as flexion to 115 degrees) after 6.7 weeks. After surgery, these patients had waited a mean of 23.6 weeks before starting home mechanical therapy. Over the course of this therapy, mean knee flexion progressed from 70.8 degrees to 130.6 degrees. Only 2 patients in this study required surgical manipulation. We conclude that a home mechanical therapy program will reduce the need for surgical management of loss of knee flexion. PMID- 12723772 TI - Cost analysis: fondaparinux versus preoperative and postoperative enoxaparin as venous thromboembolic event prophylaxis in elective hip arthroplasty. AB - Patients undergoing hip arthroplasty in the absence of prophylaxis for venous thromboembolic events (VTEs) are at high risk for experiencing postoperative VTEs. In the study reported here, we performed cost analyses involving efficacy and safety data from clinical trials evaluating fondaparinux and enoxaparin as VTE prophylaxis. Incremental cost-effectiveness ratios were calculated to determine cost per VTE avoided. In addition, cost per death averted and cost per life-year gained were calculated. Once-daily fondaparinux proved to be more cost effective than once-daily enoxaparin 40 mg but less cost-effective than twice daily enoxaparin 30 mg. PMID- 12723773 TI - Premature failure of a hip hemiarthroplasty secondary to osteolysis and aseptic loosening. AB - Bipolar hemiarthroplasty and unipolar hemiarthroplasty have been widely used, especially in cases of femoral neck fracture in elderly patients. In this article, we report on a single case of premature loosening of a bipolar endoprosthesis secondary to presence of titanium and polyethylene debris from wear. We removed the patient's prosthesis and performed a total hip arthroplasty. With use of improved surgical and cementing techniques, state-of-the-art biomaterials, and better patient selection, hemiarthroplasties recently became as long-lived a stotal hip arthroplasties. However, our patient's case of premature loosening points up some basic principles regarding the tribology of implant failure: First, the surface of titanium is poor for articulation; second, polyethylene wear, especially in cases of bipolar hemiarthroplasty with a thin liner and in the presence of titanium prostheses, may hasten failure; third, meticulous third-generation cementing techniques minimize the risk for premature failure; and fourth, hemiarthroplasties performed in patients with previous surgery have higher rates of premature failure. PMID- 12723774 TI - Bicycling-induced ulnar tunnel syndrome. AB - Symptoms of ulnar tunnel syndrome are commonly experienced by avid bicyclists. Previous orthopedic and hand surgery publications have rarely included bicycling as a causative factor for this condition. We present the case of a 41-year-old man who developed bilateral ulnar tunnel syndrome during a week-long mountain bicycling tour. His symptoms gradually improved with nonoperative treatment measures, including rest, splinting, and nonsteroidal anti-inflammatory medication. PMID- 12723775 TI - Hierarchical linear models for the quantitative integration of effect sizes in single-case research. AB - In this article, the calculation of effect size measures in single-case research and the use of hierarchical linear models for combining these measures are discussed. Special attention is given to meta-analyses that take into account a possible linear trend in the data. We show that effect size measures that have been proposed for this situation appear to be systematically affected by the duration of the experiment and fail to distinguish between effects on level and slope. To avoid these flaws, we propose to perform a multivariate meta-analysis on the standardized ordinary least squares regression coefficients from the study specific regression equations describing the response variable. PMID- 12723776 TI - Averaging learning curves across and within participants. AB - We examine recent concerns that averaged learning curves can present a distorted picture of individual learning. Analyses of practice curve data from a range of paradigms demonstrate that such concerns are well founded for fits of power and exponential functions when the arithmetic average is computed over participants. We also demonstrate that geometric averaging over participants does not, in general, avoid distortion. By contrast, we show that block averages of individual curves and similar smoothing techniques cause little or no distortion of functional form, while still providing the noise reduction benefits that motivate the use of averages. Our analyses are concerned mainly with the effects of averaging on the fit of exponential and power functions, but we also define general conditions that must be met by any set of functions to avoid distortion from averaging. PMID- 12723777 TI - Use of latent semantic analysis for predicting psychological phenomena: two issues and proposed solutions. AB - Latent semantic analysis (LSA) is a computational model of human knowledge representation that approximates semantic relatedness judgments. Two issues are discussed that researchers must attend to when evaluating the utility of LSA for predicting psychological phenomena. First, the role of semantic relatedness in the psychological process of interest must be understood. LSA indices of similarity should then be derived from this theoretical understanding. Second, the knowledge base (semantic space) from which similarity indices are generated must contain 'knowledge' that is appropriate to the task at hand. Proposed solutions are illustrated with data from an experiment in which LSA-based indices were generated from theoretical analysis of the processes involved in understanding two conflicting accounts of a historical event. These indices predict the complexity of subsequent student reasoning about the event, as well as hand-coded predictions generated from think-aloud protocols collected when students were reading the accounts of the event. PMID- 12723778 TI - An examination of daily activities and their scripts across the adult lifespan. AB - In two normative studies, we examined daily scripted activities from the perspective that scripts are frequency-based knowledge structures. In Study 1 individuals recorded their daily activities for 7 consecutive days. Fifteen activities that were reported with low, moderate, and high frequency were selected for Study 2, in which individuals generated a script for each activity. The 18 most frequently generated events from each script are reported, along with their centrality and distinctiveness rankings and the number of individuals reporting each event. Overall, the mean number of events generated increased with increasing script frequency, suggesting that script representations are subject to frequency effects. Also, we found a high level of consistency across the three age groups in the events generated in each script and in their corresponding rankings of centrality and distinctiveness. Finally, we found no evidence of age or gender bias in the frequency or recency of engaging in each of the scripted activities. PMID- 12723779 TI - Using the PLUM procedure of SPSS to fit unequal variance and generalized signal detection models. AB - The recent addition of aprocedure in SPSS for the analysis of ordinal regression models offers a simple means for researchers to fit the unequal variance normal signal detection model and other extended signal detection models. The present article shows how to implement the analysis and how to interpret the SPSS output. Examples of fitting the unequal variance normal model and other generalized signal detection models are given. The approach offers a convenient means for applying signal detection theory to a variety of research. PMID- 12723780 TI - A tool for tracking visual attention: the Restricted Focus Viewer. AB - Eye-tracking equipment has proven useful in examining the cognitive processes people use when understanding and reasoning with visual stimuli. However, eye tracking has several drawbacks: accurate eye-tracking equipment is expensive, it is often awkward for participants, it requires frequent recalibration, and the data can be difficult to interpret. We introduce an alternative tool: the Restricted Focus Viewer (RFV). This is a computer program that takes an image, blurs it, and displays it on a computer monitor, allowing the participant to see only a small region of the image in focus at any time. The region in focus can be moved using the computer mouse. The RFV records what the participant is focusing on at any point in time. It is cheap, nonintrusive, does not require calibration, and provides accurate data about which region is being focused on. We describe this tool and also provide experimental comparisons with eye-tracking. The RFV (Version 2.1) is freely available at http://www.csse.monash.edu. au/projects/RFV/. PMID- 12723781 TI - Measuring surface achromatic color: toward a common measure for increments and decrements. AB - Surface color is traditionally measured by matching methods. However, in some conditions, the color of certain surfaces cannot be measured: The surface simply looks brighter or darker than all the patches on a matching scale. We studied the reliability, validity, and range of application of three different types of simulated Munsell scales (white-, black-, and split-surrounded) as methods for measuring surface colors in simple disk-ring displays. All the scales were equally reliablefor matching both increments and decrements, but about 2096 of the increments were unmatchable on the white-surrounded scale, about 1396 of the decrements were unmatchable on the black-surrounded scale, and about 9% of the increments were unmatchable on the split-surrounded scale. However, matches on all the scales were linearly related. Therefore, it is possible to convert them to common units, using regression parameters. These units provide an extended metric for measuring all increments and decrements in the stimulus space, effectively removing ceiling and floor effects, and providing measures even for surfaces that were perceived as out of range on some of the scales. PMID- 12723782 TI - A simple system for the remote detection and analysis of sniffing in explosives detection dogs. AB - In this article, we describe a device that we have developed, for both research and operational purposes, to assist handlers of sniffer dogs and improve efficiency of search. The device transmits sounds from the dog's muzzle area to a listener, enabling determination of whether the dog is actually sniffing during a search. Both the aural and the computerized sound analyses enable study of the dog's sniffing behavior under a variety of situations. Correlation of these data with the visually monitored behavior of the working dog contributes additional and important information and leads to a greater understanding of detector dogs' behavior and abilities. PMID- 12723783 TI - The palm IAT: a portable version of the implicit association task. AB - The Implicit Association Test (IAT) uses reaction times to measure implicit linkages among concepts or between concepts and attributes. The Palm IAT is a simplified version of the IAT that runs on Palm Pilot or Handspring Visor personal digital assistant devices. The Palm IAT is portable, provides precise measurement, and allows for efficient data collection inside or outside the laboratory. Three studies were conducted to asses its practical usefulness. Study 1 (N = 1 2) replicated a previous demonstration of more favorable implicit associations for flowers than for insects. Study 2 (N = 9) found individual differences in linking the self-concept to characteristics of agency and communion. Study 3 (N = 23) found individual differences in linking the self concept to characteristics of boldness and friendliness. Reliability of the Palm IAT is similar to reliability of theoriginal desktop computer version. Using the Palm IAT, one can study hard-to-reach populations in unusual settings. PMID- 12723784 TI - Timing accuracy under microsoft windows revealed through external chronometry. AB - Recent studies by Myors (1998, 1999) have concluded that the Microsoft Windows operating system is unable to support sufficient timing precision and resolution for use in psychological research. In the present study, we reexamined the timing accuracy of Windows 95/98, using (1) external chronometry, (2) methods to maximize the system priority of timing software, and (3) timing functions with a theoretical resolution of 1 msec or better. The suitability of various peripheral response devices and the relative timing accuracy of computers with microprocessors with different speeds were also explored. The results indicate that if software is properly controlled, submillisecond timing resolution is achievable under Windows with both old and new computers alike. Of the computer input devices tested, the standard parallel port was revealed as the most precise, and the serial mouse also exhibited sufficient timing precision for use in single-interval reaction time experiments. PMID- 12723785 TI - A simple and sensitive method to measure timing accuracy. AB - Timing accuracy in presenting experimental stimuli (visual information on a PC or on a TV) and responding (keyboard presses and mouse signals) is of importance in several experimental paradigms. In this article, a simple system for measuring timing accuracy is described. The system uses two PCs (at least Pentium II, 200 MHz), a photocell, and an amplifier. No additional boards and timing hardware are needed. The first PC, a SlavePC, monitors the keyboard presses or mouse signals from the PC under test and uses a photocell that is placed in front of the screen to detect the appearance of visual stimuli on the display. The software consists of a small program running on the SlavePC. The SlavePC is connected through a serial line with a second PC. This MasterPC controls the SlavePC through an ActiveX control, which is used in a Visual Basic program. The accuracy of our system was investigated by using a similar setup of a SlavePC and a MasterPC to generate pulses and by using a pulse generator card. These tests revealed that our system has a 0.01-msec accuracy. As an illustration, the reaction time accuracy of INQUISIT for a few applications was tested using our system. It was found that in those applications that we investigated, INQUISIT measures reaction times from keyboard presses with millisecond accuracy. PMID- 12723786 TI - DMDX: a windows display program with millisecond accuracy. AB - DMDX is a Windows-based program designed primarily for language-processing experiments. It uses the features of Pentium class CPUs and the library routines provided in DirectX to provide accurate timing and synchronization of visual and audio output. A brief overview of the design of the program is provided, together with the results of tests of the accuracy of timing. The Web site for downloading the software is given, but the source code is not available. PMID- 12723787 TI - Presenting multiple auditory signals using multiple sound cards in Visual Basic 6.0. AB - In auditory research, it is often desirable to present more than two auditory stimuli at any one time. Although the technology has been available for some time, the majority of researchers have not utilized it. This article provides a simple means of presenting multiple, concurrent, independent auditory events, using two or more different sound cards installed within a single computer. By enabling the presentation of more auditory events, we can hope to gain a better understanding of the cognitive and attentional processes operating under more complex and realistic scenes, such as that embodied by the cocktail party effect. The software requirements are Windows 98SR2/Me/NT4/2000/XP, Visual Basic 6.0, and DirectX 7.0 or above. The hardware requirements are a Pentium II, 128 MB RAM, and two or more different sound cards. PMID- 12723788 TI - Generating anagrams from multiple core strings employing user-defined vocabularies and orthographic parameters. AB - Anagrams are used widely in psychological research. However, generating a range of strings with the same letter content is an inherently difficult and time consuming task for humans, and current computer-based anagram generators do not provide the controls necessary for psychological research. In this article, we present a computational algorithm that overcomes these problems. Specifically, the algorithm processes automatically each word in a user-defined source vocabulary and outputs, for each word, all possible anagrams that exist as words (or as nonwords, if required) as defined by the same source vocabulary. Moreover, we show how the output of the algorithm can be filtered to produce anagrams within specific user-defined orthographic parameters. For example, the anagrams produced can be filtered to produce words that share, with each other or with other words in the source vocabulary, letters in only certain positions. Finally, we provide free access to the complete Windows-based program and source code containing these facilities for anagram generation. PMID- 12723789 TI - Grapheme-phoneme probabilities in British English. AB - A computational analysis of a large British English database was performed and frequencies occurrence of grapheme-phoneme correspondences were obtained. A computer program was implemented, which used these frequencies to predict the probabilities of all possible pronunciations of any given string of graphemes. These results led to a proposal for a quantitative method of measurement of the orthographic depth of different languages. PMID- 12723790 TI - A new set of 299 pictures for psycholinguistic studies: French norms for name agreement, image agreement, conceptual familiarity, visual complexity, image variability, age of acquisition, and naming latencies. AB - Pictures are often used as stimuli in studies of perception, language, and memory. Since performances on different sets of pictures are generally contrasted, stimulus selection requires the use of standardized material to match pictures across different variables. Unfortunately, the number of standardized pictures available for empirical research is rather limited. The aim of the present study is to provide French normative data for a new set of 299 black-and white drawings. Alario and Ferrand (1999) were closely followed in that the pictures were standardized on six variables name agreement, image agreement, conceptual familiarity, visual complexity, image variability, and age of acquisition. Objective frequency measures are also provided for themost common names associated with the pictures. Comparative analyses between our results and the norms obtained in other, similar studies are reported. Finally, naming latencies corresponding to the set of pictures were also collected from French native speakers, and correlational/multiple-regression analyses were performed on naming latencies. This new set of standardized pictures is available on the Internet (http://leadserv.u-bourgogne.fr/bases/pictures/) and should be of great use to researchers when they select pictorial stimuli. PMID- 12723791 TI - Normative data and naming times for action pictures. AB - The present article provides Spanish norms for name agreement, printed word frequency, word compound frequency, familiarity, imageability, visual complexity, age of acquisition, and word length (measured by syllables and phonemes) for 100 line drawings of actions taken from Druks and Masterson (2000). In addition, through a naming-time experiment carried out with a group of 54 Spanish students in a pool of 63 of these line drawings, we determined the best predictors of naming actions. In the multiple regression analysis, age of acquisition and name agreement emerged as the most important determinants of action-naming reaction time. PMID- 12723792 TI - The global methane cycle: isotopes and mixing ratios, sources and sinks. AB - A review of the global cycle of methane is presented with emphasis on its isotopic composition. The history of methane mixing ratios, reconstructed from measurements of air trapped in ice-cores is described. The methane record now extends back to 420 kyr ago in the case of the Vostok ice cores from Antarctica. The trends in mixing ratios and in delta13C values are reported for the two Hemispheres. The increase of the atmospheric methane concentration over the past 200 years, and by 1% per year since 1978, reaching 1.7 ppmv in 1990 is underlined. The various methane sources are presented. Indeed the authors describe the methane emissions by bacterial activity under anaerobic conditions in wet environments (wetlands, bogs, tundra, rice paddies), in ruminant stomachs and termite guts, and that originating from fossil carbon sources, such as biomass burning, coal mining, industrial losses, automobile exhaust, sea floor vent, and volcanic emissions. Furthermore, the main sinks of methane in the troposphere, soils or waters via oxidation are also reported, and the corresponding kinetic isotope effects. PMID- 12723793 TI - Softly, softly: veterinarians and conservation practioners working in the developing world. PMID- 12723794 TI - Pharmacokinetics of florfenicol in loggerhead sea turtles (Caretta caretta) after single intravenous and intramuscular injections. AB - The pharmocodynamics of single injections of florfenicol in yearling loggerhead sea turtles (Caretta caretta) were determined. Eight juvenile loggerhead sea turtles weighing 1.25 (+/- 0.18) kg were divided into two groups. Four animals received 30 mg/kg of florfenicol i.v., and four received the same dose i.m. Plasma florfenicol concentrations were analyzed by reverse-phase high performance liquid chromatography. After the i.v. dose, there was a biphasic decline in plasma florfenicol concentration. The initial steep phase from 3 min to 1 hr had a half-life of 3 min, and there was a longer slow phase of elimination, with a half-life that ranged from 2 to 7.8 hr among turtles. The volume of distribution varied greatly and ranged from 10.46 to -60 L/kg. Clearance after the i.v. dose was 3.6-6.3 L/kg/hr. After the i.m. injection, there was a peak within 30 min of 1.4-5.6 microg/ml, and florfenicol was thereafter eliminated with a half-life of 3.2-4.3 hr. With either route, florfenicol plasma concentrations were below the minimum inhibitory concentrations for sensitive bacteria within 1 hr. Florfenicol does not appear to be a practical antibiotic in sea turtles when administered at these doses. PMID- 12723795 TI - Disposition of sulfadimethoxine in male llamas (Llama glama) after single intravenous and oral administrations. AB - This study determined the disposition of sulfadimethoxine in six, healthy, adult, gelded male llamas (Llama glama) by using a nonrandomized crossover design with i.v. dosing (58.8 +/- 3.0 mg/kg based on metabolic scaling) followed by oral dosing (59.3 mg/kg +/- 8.3). Blood samples were collected intermittently for a 72 hr period, and serum sulfadimethoxine concentrations were quantified using high performance liquid chromatography. Serum sulfadimethoxine concentrations across time were subjected to standard pharmacokinetic analysis based on linear regression. Mean maximum serum concentration after oral dosing was 23.6 +/- 14.9 microg/ml, and extrapolated peak concentration after i.v. administration was 246.6 +/- 15.8 microg/ml. Total clearance of sulfadimethoxine was 45.4 +/- 13.9 L/kg. Half-lives after i.v. and oral administration were 541 +/- 111 min and 642.4 +/- 204.8 min, respectively. Oral bioavailability was 52.6 +/- 15%. These data suggest that the oral dose administered to llamas in this study, based on metabolic scaling from cattle, may be inadequate when compared with the reported minimum inhibitory concentration (512 microg/ml) breakpoint for sulfadimethoxine. PMID- 12723796 TI - Biomedical evaluation of free-ranging ring-tailed lemurs (Lemur catta) in Tsimanampetsotsa Strict Nature Reserve, Madagascar. AB - Complete health assessments were performed on 20 adult ring-tailed lemurs (Lemur catta), 10 males and 10 females, free ranging within the Tsimanampetsotsa Strict Nature Reserve in southwest Madagascar. Each animal was anesthetized using tiletamine and zolazepam, weighed, and given a thorough physical examination. Blood was collected for hematology, determination of plasma total protein concentration, serum chemistry, determination of fat-soluble vitamin and trace mineral concentrations, assessment of iron metabolism, toxoplasmosis and viral serologies, and examination for parasites. Feces were collected for bacterial culture and parasite examination, and representative numbers of ectoparasites were collected. Blood values differed significantly in a number of ways from values in captive lemurs, possibly associated with recent food consumption, hydration, and diet. There was no evidence of serious infectious disease and no hemoparasites. The enteric flora appeared unremarkable, although results may have been skewed toward cold-tolerant bacteria. The fecal parasite burden was low, but there were large numbers of ectoparasites (mites) present. Our study demonstrated that a substantial amount of data can be collected from free-ranging populations, information that is invaluable in the management of captive populations, particularly with regard to disease, and in assessing risks associated with reintroduction programs. PMID- 12723797 TI - Evaluation of oral and subcutaneous delivery of an experimental canarypox recombinant canine distemper vaccine in the Siberian polecat (Mustela eversmanni). AB - We assessed the safety and efficacy of an experimental canarypox-vectored recombinant canine distemper virus (CDV) subunit vaccine in the Siberian polecat (Mustela eversmanni), a close relative of the black-footed ferret, (M. nigripes), an endangered species that is highly susceptible to the virus. Siberian polecats were randomized into six treatment groups. Recombinant canine distemper vaccine was administered s.c. at three dose levels (10(4.5), 10(5.0), and 10(5.5) plaque forming units [PFU] per dose) and was administered orally by spraying the vaccine into the oropharnyx at two dose levels (10(5.5), 10(8.0) PFU per dose). The sixth group of control animals was not vaccinated. For both routes of administration, two 1-ml doses of reconstituted vaccine were delivered 4 wk apart, followed by live virus challenge 3 wk after the second vaccination. During the challenge, Synder Hill test strain CDV obtained from the National Veterinary Services Laboratory in Ames, Iowa, was administered i.p. Serial blood samples for CDV serology were collected immediately before vaccination and challenge, and 10, 15, and 20 days after challenge. Clinical signs and body weights were recorded up to 32 days after challenge. The survival rate in animals receiving vaccine at the highest oral dose (10(8.0) PFU per dose) was 83.3%. Survival rate was 50.0% in the high s.c. and 60.0% in the medium s.c. groups. All animals in the low-s.c. dose, low-oral dose, and control groups died after exposure. Vaccine dose overall (oral and s.c.) and dose in response to s.c. administration when considered alone were significant predictors of survival (P = 0.006 and P = 0.04, respectively). Among the polecats challenged with virulent virus, those that died became sick sooner than those that survived. Animals that died lost significantly more weight during the 10 days after challenge than did animals that survived (P = 0.02). Survival rates did not differ by sex, founder female status, or breeding pedigree in any of the treatment groups. Survival rates were higher in animals with increasing serum neutralization titers (P = 0.027). This study demonstrates the efficacy of oral delivery of a recombinant CDV vaccine in the Siberian polecat. Further studies are needed to evaluate the safety and efficacy of vectored recombinant vaccines in highly susceptible species and especially in those species in which vaccination with modified live CDV has led to disease. PMID- 12723798 TI - Idiopathic acute onset myelopathy in cheetah (Acinonyx jubatus) cubs. AB - Numerous cases of ataxia, hind limb paresis, and paralysis have occurred in cheetah (Acinonyx jubatus) cubs over the past 10 yr within the European Endangered Species Program population, including 12 in mainland Europe, two in the British Isles, one in Namibia, and one in Dubai. The condition is the most important medical factor limiting European cheetah population growth. Eight cubs at the Salzburg Zoo, Austria, were affected. They demonstrated upper motor neuron lesions when alive and bilateral, symmetrical myelin degeneration of the spinal cord on necropsy. Ballooning of myelin sheaths surrounded mostly preserved axons, and no spheroids, characteristic of acute axonal degeneration, were found. Myelin loss markedly exceeded axonal degeneration. The syndrome's etiology is unclear, although viral, bacterial, parasitic, genetic, nutritional-metabolic, toxic, and physical causes have been considered. PMID- 12723799 TI - Comparison of the cardiorespiratory effects of medetomidine-butorphanol-ketamine and medetomidine-butorphanol-midazolam in patas monkeys (Erythrocebus patas). AB - The cardiorespiratory effects, effectiveness, and reversibility of two injectable anesthetic combinations were compared in captive patas monkeys (Erythrocebus patas). Seven patas monkeys were hand-injected with medetomidine (40 microg/kg, i.m.), butorphanol (0.4 mg/kg. i.m.), and ketamine (3.0 mg/kg. i.m.), and seven were injected with the same dosages of medetomidine and butorphanol plus midazolam (0.3 mg/kg, i.m.). Heart rates decreased in monkeys in both treatment groups and were lower than those previously recorded in patas monkeys anesthetized with either ketamine or ketamine and isoflurane. Mean arterial pressures were highest in ketamine-treated monkeys but remained within normal limits for both groups. End tidal CO2 values increased gradually over time in both groups and were above physiologic norms after 20 min. Respiratory rates were similar between groups and remained constant throughout the procedures. Despite adequate ventilation parameters, initial low percent oxygen-hemoglobin saturation (SpO2) values were suggestive of severe hypoxemia. It was not clear whether these were accurate readings or an artifact of medetomidine-induced peripheral vasoconstriction. Oxygen supplementation restored SpO2 values to normal (>94%) in both groups. Both combinations effectively produced a state of light anesthesia, although spontaneous recoveries occurred after 30 min in three ketamine-treated monkeys. All monkeys were given i.m. atipamezole (0.2 mg/kg) and naloxone (0.02 mg/kg), whereas midazolam-treated monkeys also received flumazenil (0.02 mg/kg, i.v.), which resulted in faster (median recovery time = 5 min) and more complete recoveries in this group. Both combinations are safe to use when supplemented with oxygen, although the midazolam combination provided a longer anesthetic period and was more fully reversible. PMID- 12723800 TI - A comparison between pre- and posthibernation morphometry, hematology, and blood chemistry in viperid snakes. AB - Snakes from temperate climates are often made to hibernate in zoos to stimulate reproduction. Unfortunately, deaths have occurred during and after hibernation. This study evaluated the health status, pre- and posthibernation, of 31 adult viperid snakes. It included morphometric measurements, hematology, and blood chemistry. No differences were seen in body weights and weight to length ratios between pre- and posthibernation examinations, suggesting that the overall condition of the snakes did not change. No differences were seen in hematologic and blood chemistry parameters, except that bile acids (3alpha-hydroxybile acids) decreased, the implications of which are unknown. Three individuals had markedly high plasma uric acid levels posthibernation; of these, two individuals died from extensive visceral gout and one recovered with fluid therapy. Viperid snakes should be clinically healthy, well hydrated, and in good body condition when they are put into hibernation. They should be maintained in an environment with sufficient humidity and should have access to water. Blood samples should be collected on arousal for measuring plasma uric acid levels. Changes in morphometry, hematology, and blood chemistry appear to be abnormal and should be investigated thoroughly. PMID- 12723801 TI - Effect of venipuncture site and anticoagulant on selected hematologic values in black rhinoceros (Diceros bicornis). AB - Paired blood samples were collected from the ear and radial vein of four captive healthy adult black rhinoceroses (Diceros bicornis). Samples were collected using heparin or ethylenediaminetetraacetic acid (EDTA) as an anticoagulant. Packed cell volume (PCV) and total protein (TP) values were compared between samples drawn from the two venipuncture sites and treated with the two anticoagulants to determine whether statistically significant variation occurred. No significant difference in the grouped values was observed when venipuncture sites (ear and radial vein) were compared using the same anticoagulant (heparin). However, when comparing different anticoagulants (EDTA and heparin) used to collect blood from the radial vein, the grouped-heparinized samples had higher mean PCV and TP values than did the EDTA-treated samples. These differences may be important when performing serial sampling in a sick rhinoceros and suggest that the choice of anticoagulant should be consistent, although selection of venipuncture site may be less important when monitoring selected hematologic values in black rhinoceroses. PMID- 12723802 TI - Serosurvey for feline leukemia virus and lentiviruses in captive small neotropic felids in Sao Paulo state, Brazil. AB - Feline leukemia virus (FeLV), Gammaretrovirus, and feline immunodeficiency virus, a Lentivirus, are members of the family Retroviridae, and may establish persistent infections in the domestic cat (Felis catus). Cytoproliferative and cytosuppressive disorders may result from infection with these viruses. Morbidity and mortality rates are high in domestic cats worldwide. Infection of endangered neotropic small felids with these viruses could be devastating. To investigate the prevalence of FeLV and feline lentiviruses in neotropic small felids kept in captivity in Sao Paulo state. Brazil, serum samples from 104 animals belonging to the species Leopardus pardalis, Leopardus tigrinus, Leopardus wiedii, Herpailurus yaguarondi, and Oncifelis geoffroyi were tested for FeLV and feline lentiviruses by commercially available immunoassays. All results were negative, suggesting that retrovirus infection is not an important clinical problem in these populations. Because domestic cats in Sao Paulo city are naturally infected with these pathogens, and feral cats are commonly found in zoologic facilities in Brazil, preventive measures should be taken to avoid transmission of retroviruses to naive populations of wild and captive neotropic felids in Brazil. PMID- 12723803 TI - Proliferative enteritis in leopard geckos (Eublepharis macularius) associated with Cryptosporidium sp. infection. AB - Twenty-three leopard geckos (Eublepharis macularius) with various clinical histories of weight loss, anorexia, lethargy, and diarrhea were submitted either intact or as biopsy specimens to the University of Florida Anatomic Pathology Service. Gross necropsy findings in the intact geckos included marked reduction of subcutaneous adipose tissue stores at the tail base and mild thickening and reddening of the small intestine. Histologic examination revealed Cryptosporidium sp. infection associated with hyperplasia and mononuclear inflammation of the small intestine in all geckos. Parasites and lesions were only rarely observed in the stomach and large intestine of geckos. The histologic and ultrastructural lesions in the small intestine of leopard geckos infected with Cryptosporidium sp. have not been well characterized previously. This report implicates Cryptosporidium sp. as the cause of disease in the geckos and describes the range of histologic lesions observed. PMID- 12723804 TI - A molecular epidemiologic investigation of Salmonella from a meat source to the feces of captive cheetah (Acinonyx jubatus). AB - Low cheetah (Acinonyx jubatus) birth rates were observed for a long time in a captive breeding facility in which Salmonella, which was possibly present in contaminated beef, was isolated from still-born lion (Panthera leo) cubs. Salmonella, including 14 isolates of Salmonella serovar typhimurium and 19 isolates of Salmonella serovar muenchen, was subsequently isolated 47 times from 378 meat samples at the facility during a 13-mo period. Salmonella, including 26 isolates of S. serovar typhimurium, 10 of S. serovar muenchen, and 11 other serovars, also was isolated 54 times from 119 fecal samples. Only three plasmid profiles were identified in 59 S. typhimurium isolates from both meat and fecal samples. Although random-amplified polymorphic DNA fingerprinting using different primers in the polymerase chain reaction was able to distinguish between S. typhimurium and S. muenchen and to demonstrate similar chromosomal DNA fingerprints in some of the isolates from meat and feces, the results were not consistent enough to prove that the Salmonella in the feces originated from contaminated meat. However, the predominance of only two serovars in the meat fed to carnivores and in the feces of these animals suggests that the meat was the source of the Salmonella organisms in the feces. PMID- 12723805 TI - Oral treatment of avian lead intoxication with meso-2,3-dimercaptosuccinic acid. AB - The efficacy of meso-dimercaptosuccinic acid (DMSA) (succimer) in treating avian lead intoxication was studied in a retrospective, nonrandomized, longitudinal study. Nineteen birds with moderate to high blood lead concentration and neurologic signs compatible with lead toxicity were treated with DMSA (30 mg/kg p.o., b.i.d.; n = 15) for a minimum of 7 days. In cases with severe neurologic signs, DMSA was supplemented with a single dose of edetate calcium disodium (<50.0 mg/kg of body weight i.m.; n = 4). Blood lead concentrations were measured two or more times (before and after treatment). Median blood lead concentration decreased (87%), neurologic signs were resolved, and there were no apparent adverse secondary effects. PMID- 12723806 TI - Endoscopic evaluation of the esophagus and stomach in three loggerhead sea turtles (Caretta caretta) and a Malaysian giant turtle (Orlitia borneensis). AB - Three loggerhead sea turtles (Caretta caretta) and a Malaysian giant turtle (Orlitia borneensis) were presented with suspected or confirmed esophageal foreign bodies. Esophagoscopy was performed on all turtles, and gastroscopy was performed on three turtles. In all cases, endoscopy was easy to perform, and allowed visualization of most upper gastrointestinal features. The papillated esophagus was easy to navigate, but mucosal papillae in the loggerhead sea turtles prevented examination of the underlying mucosa. The stomach was easily entered and examined in both species, but the working endoscope length (100 cm) prevented inspection of the pyloric antrum and the duodenum in all turtles. The turtles in this report may serve as references for future endoscopic examinations of these species. PMID- 12723807 TI - Induced elephant (Loxodonta africana) tusk removal. AB - Elephant tusk removal usually requires costly surgical procedures that are time consuming and present a significant risk to the animal when performed using general anesthesia. Such techniques require gauges, chisels, and forceps to remove the tusk. This article reports the simple removal of the tusk of an 18-yr old African elephant (Loxodonta africana) without the use of surgical instruments and anesthesia. Rubber elastics were placed around a tusk, causing loss of alveolar bone with subsequent exfoliation of the tusk within 3 wk. The healing process was uneventful. PMID- 12723808 TI - Klinefelter syndrome (39 XXY) in an adult Siberian tiger (Panthera tigris altaica). AB - Fibroblast cultures of a skin biopsy from an adult intact male Siberian tiger (Panthera tigris altaica) revealed an abnormal standard and G-banded karyotype diploid chromosome number of 2n = 39 XXY due to an extra sex chromosome as opposed to the expected 2n = 38 XY. The tiger was euthanatized 1 yr later due to acute multifocal intervertebral disc disease. Histopathology of the reproductive tract demonstrated a paucity of seminiferous tubules and these were devoid of spermatagonia. An increase in fibrous connective tissue was noted in sections of the prostate and epididymis, and expansion of the fibrous interstitium was observed in the testes. PMID- 12723809 TI - Visceral mast cell tumor in a captive black jaguar (Panthera onca). AB - Little is known about neoplasia in the jaguar (Panthera onca), the largest American feline. A captive black jaguar was diagnosed at necropsy with a mastocytic form of visceral mast cell tumor similar to that which occurs in domestic cats. This animal had no previous clinical disease and died during anesthesia for a routine dental treatment. PMID- 12723810 TI - Chat reference service in medical libraries: part 1--An introduction. AB - Chat reference services offer the opportunity to provide immediate access to quality information to meet the medical information needs of students, faculty, staff, physicians, nurses, and allied health care professionals. Part 1 of this two-part article on chat reference service in medical libraries is an introduction to the management of chat reference services and to features available for chat reference. The management of chat reference services raises issues of planning, staffing, selecting, and marketing. Planning issues focus on the identification of the users, the users' medical information needs, and the users' information-seeking behavior. Staffing issues relate to the selection of chat hours, the location of the chat service, and participation in collaborative agreements. Selecting chat software weighs the sophistication of the chat features against the related cost. Marketing uses techniques similar to traditional reference services and often begins slowly as chat expertise develops. Part 2 of the article discusses trends in chat reference services in medical libraries. PMID- 12723812 TI - The Region 4 collaborative virtual reference project. AB - In May 2002, the Denison Memorial Library at the University of Colorado Health Sciences Center and the J. Otto Lottes Health Sciences Library at the University of Missouri-Columbia, with funding from the National Network of Libraries of Medicine-Midcontinental Region, embarked on a collaborative, real-time reference project using the 24/7 Reference, Inc., software package. This paper describes how the project was conceived, and includes details on the service hours, staffing, training, marketing, lessons learned, and future plans for the service. PMID- 12723811 TI - Chat reference service in medical libraries: part 2--Trends in medical school libraries. AB - An increasing number of medical school libraries offer chat service to provide immediate, high quality information at the time and point of need to students, faculty, staff, and health care professionals. Part 2 of Chat Reference Service in Medical Libraries presents a snapshot of the current trends in chat reference service in medical school libraries. In late 2002, 25 (21%) medical school libraries provided chat reference. Trends in chat reference services in medical school libraries were compiled from an exploration of medical school library Web sites and informal correspondence from medical school library personnel. Many medical libraries are actively investigating and planning new chat reference services, while others have decided not to pursue chat reference at this time. Anecdotal comments from medical school library staff provide insights into chat reference service. PMID- 12723813 TI - Promotion and evaluation of a virtual live reference service. AB - The University of North Carolina at Chapel Hill Health Sciences Library began providing virtual live reference service in the fall of 2001. The library was interested in exploring new ways to interact with users accessing electronic resources from outside the library. This article describes the background for starting the service, the methods used to promote the service, and the results of an evaluation conducted after the first seven months of operation. The service was lightly used during these months with only 82 interactions. An analysis of the transcripts examined referring Web pages, types of questions asked, software features used, and user login data. Text of two user surveys is included, with the data from the in-person survey analyzed. PMID- 12723814 TI - Not virtual, but a real, live, online, interactive reference service. AB - In today's fast-paced environment, traditional medical reference services alone are not adequate to meet users' information needs. Efforts to find new ways to provide comprehensive service to users, where and when needed, have often included the use of new and developing technologies. This paper describes the experience of an academic health science library in developing and providing an online, real-time reference service. Issues discussed include selecting software, training librarians, staffing the service, and considering the future of the service. Use statistics, question type analysis, and feedback from users of the service and librarians who staff the service, are also presented. PMID- 12723815 TI - Planning, implementing, and using a virtual reference service. AB - Virtual reference (VR) can be a successful vehicle for libraries if there is a demonstrated need by the user base for such a service and if the library staff believes in, accepts, and plans thoroughly for the concept. This article focuses on the experiences of the Duke University Medical Center Library (DUMCL) in planning, implementing, and using a virtual reference service utilizing LSSI's Virtual Reference Desk (VRD) Software. PMID- 12723816 TI - PD(X)MD: a new clinical information tool for patient care. AB - PD(X)MD is a new and promising evidence-based electronic primary care clinical information resource. It is geared primarily to the physician, as is its companion database, MDConsult, but it can also be very useful to medical and non medical librarians seeking to quickly summarize current, accurate, and quality filtered clinical information focusing on diagnosis, therapy, prognosis, and prevention. PMID- 12723817 TI - One library's experience with review and selection of chat software for reference. AB - When Michigan State University (MSU) Libraries decided to make the foray into virtual reference, the first thing that needed to be done was to decide on the software to use. This article discusses the process used including the items considered essential (deal-breakers) for software to make the first cut, what other features needed to be included, and what features would be useful but were not critical. A literature review of some useful current articles on virtual reference is included. The vendor and software ultimately selected was not one of the original vendors; how MSU Libraries was able to evaluate and select Docutek is presented. A matrix for software comparison is included in the appendix. PMID- 12723818 TI - "Ask A Librarian": the experiences of four hospital libraries. AB - The librarians at four hospital libraries describe the electronic reference service, "Ask A Librarian," offered at their institutions. The hospitals are vastly different in size and in number of library staff, and offer the "Ask A Librarian" service to different clientele. The article illustrates that both large hospitals with a large library staff and small hospitals with a solo librarian and some volunteers can offer this type of service. PMID- 12723820 TI - What's this HIPAA stuff ... and does it affect me? PMID- 12723819 TI - Teaching EBM teachers: a team approach. AB - A collaborative Evidence-Based Medicine (EBM) course, co-taught by medical and library faculty, targets the needs of health care and information professionals involved in teaching and practicing EBM by introducing concepts, methods and tools of the EBM process. The one-week course engages working teams of health care and information professionals in case-based activities to encourage participation in the learning process and the "Train the Trainer" approach of the course. The interdisciplinary partnership, both of teachers and learners, has led to a successful and effective educational program that accentuates the strengths and contributions of each group. Further research will determine whether this collaborative experience leads to ongoing partnerships between these professional groups in the clinical setting. PMID- 12723821 TI - The ABCs of pediatric laboratory interpretation: understanding the CBC with differential and LFTs. AB - Interpretation of pediatric laboratory values is an essential component of comprehensive nursing care delivery to children and their families. Key laboratory interpretation strategies for common pediatric laboratory values including the complete blood count (CBC) with differential and liver function tests (LFTs) are discussed. Basic anatomy and physiology, normal laboratory parameters, and rationale for deviations from normal are reviewed. PMID- 12723822 TI - Pediatric and neonatal cardiovascular pharmacology. AB - Advances in cardiology, surgical techniques, postoperative care, and medications have improved the chances of long-term survival of the neonatal and pediatric patient with complex congenital cardiac anomalies. Rather than undergoing palliative repair, these children are now frequently taken to the operating room for complete repair. As complete repair becomes the norm, collaborative management and a thorough understanding of the pre and postoperative medications used become essential to the care of these patients. The nurse's ability to understand preop, postop, and management medications is enhanced by an understanding of the principles of cardiac anatomy and physiology, as well as developmental changes in cardiac function. All of these are reviewed. In addition, since the safe and effective administration of these drugs depends on the pediatric intensive care unit (PICU) and neonatal intensive care unit (NICU) nurse's thorough knowledge of these medications and their effects on the cardiovascular system, a brief review of these medications is presented. While new technology and techniques are improving survival rates for children with congenital heart anomalies, it is the postoperative care that these children receive that enhances the patient's survival even more. PMID- 12723823 TI - Evaluation of a new, wearable, precision phase-change thermometer in neonates. AB - The primary purpose of this research was to examine the quantitative relationship between a new, wearable, continuous-read, precision phase-change thermometer (WCPPT) and a glass-mercury thermometer used in the axilla (GMA) in neonates. The study design was prospective and quasi-experimental with each subject acting as his/her own control. Data were collected in a tertiary teaching facility in the Southwestern United States from a convenience sample of 29 neonates. Simultaneous test and reference temperature measurements were taken from each neonate. Measurements were compared to determine clinical bias (mean difference +/- 1 standard deviation [SD]) and clinical agreement portrayed graphically (mean difference and limits of agreement plotted against the mean of both test and reference measurements) (Bland & Altman, 1986). Key findings included a clinical bias between the GMA and the TraxIt WCPPT of 0.04 degrees C (+/- 0.22) (mean difference +/- 1 SD) during the first measurement session with the GMA slightly higher, and -0.11 degrees C (+/- 0.17) (mean difference +/- 1 SD) during the second measurement with the WCPPT slightly higher. Bland and Altman representations supported these findings. These outcomes support a conclusion that the TraxIt WCPPT is neither clinically nor statistically different from the GMA on initial placement and exceeds GMA readings during prolonged, continuous axillary contact. Continuous thermometer placement eliminates drawdown, the transitory, local cooling effect of intermittent thermometer placement. PMID- 12723824 TI - Anemia: when is it iron deficiency? AB - Iron deficiency anemia, the most common of childhood anemias, presents many challenges to the practitioner. Careful history taking that includes nutritional assessment may uncover this frequently subtle condition. In keeping with AAP recommendations, screening will aid in diagnosing this condition in populations at risk. Prompt treatment and conscientious follow-up will afford the most optimal outcome. PMID- 12723825 TI - Pediatric management problems. PMID- 12723827 TI - Say when ... end-of-life decisions in PICU. PMID- 12723826 TI - Facing challenges to family-centered care. I: Conflicts over visitation. AB - Policies and practices related to parental presence and participation in the care of a hospitalized child can be a source of tension between nurses and families. Tensions often revolve around "visiting" hours, who may visit, the number of visitors allowed at the bedside at a time, and inconsistent enforcement of existing visitation policies. A family-centered framework for evaluating these policies and practices can provide direction that will help reduce these tensions. Visitation policies that are flexible and offer guidelines, not rules, will usually best meet the needs of families. Some nurses may need education, mentoring, skill-building, and role-playing opportunities to work comfortably with flexible guidelines and increased family presence and participation. PMID- 12723828 TI - Clinical validation of FLACC: preverbal patient pain scale. AB - PURPOSE: To test the validity of the Faces, Legs, Activity, Cry and Consolability (FLACC) pain assessment tool by measuring changes in scores in response to analgesics. METHOD: METHODS: Pediatric nurses used the FLACC scale to assess pain in 147 children under 3 years of age who were hospitalized in the pediatric intensive care unit (PICU), post-anesthesia care unit (PACU), surgical/trauma unit, hematology/oncology unit, or infant unit. FLACC is an observational tool for quantifying pain behaviors. Facial expression, leg movement, activity, cry, and consolability are each scored 0-2, for a total FLACC score of 0-10. The FLACC measurements were done pre-analgesia, at predicted onset of analgesia, and at predicted peak analgesia. FINDINGS: Pre-analgesia FLACC scores were significantly higher than post-analgesic scores and significantly higher for patients who received opioids than patients who received non-opioids. Peak analgesia FLACC scores across analgesia groups were not significantly different and reflect effective pain relief for patients regardless of analgesic choice. CONCLUSIONS: The FLACC pain assessment tool is appropriate for preverbal children in pain from surgery, trauma, cancer, or other disease processes. The results support pediatric nurses' clinical judgment to determine analgesic choice rather than providing distinct FLACC scores to guide analgesic selection. PMID- 12723829 TI - Finding and appraising systematic reviews of clinical interventions: critical skills for evidence-based practice. PMID- 12723830 TI - Ten things I learned while writing my last research grant. AB - There are nurses in numerous roles today fulfilling their professional responsibilities by engaging in a variety of clinical, educational, administrative and research activities. Nurse researchers strive to develop the knowledge base that nurse clinicians use in practice, and their synergistic efforts contribute toward continuously improving patient care. For nurse clinicians, the daily pursuit of excellence is evidenced in their rewards for exemplary clinical performance; for nurse researchers, receiving recognition and funding from the National Institutes of Health (NIH) is the hallmark of their accomplishments. But seeking research grant money is an arduous and complex ordeal requiring focus, persistence and deliberate action. Many seek the pinnacle "RO1"--that research grant which endorses one's scientific status in the nursing research community--and many never succeed. This author offers advice to those who would not be weak in heart or courage--to attempt to climb this pinnacle. PMID- 12723831 TI - Compliance leadership: the 17th century model that doesn't work. AB - The world of health care has changed. We can't operate on 17th century models and be successful. We don't have to argue for the movement to committed, inspired leadership models, and the death of compliance leadership. There is abundant research to document this is the way we must go. In reality, we have a moral obligation to provide the kind of work environment that provides the meaningful work that Maslow (1998) tells us makes life meaningful. No one has the right to make people miserable at work because we have failed to create the cultures that create commitment, inspiration, and transformation for our patients, their families, and our staff. It is unfortunate that in times of staff shortages, this message is heard louder. We should be equally committed no matter what the situation, because it is the right thing to do. We do not have the right to be abusive to others in any context. PMID- 12723832 TI - Two-year outcomes of primary molar ferric sulfate pulpotomy and root canal therapy. AB - PURPOSE: The objective of this outcome study was to compare ferric sulfate pulpotomy (FS) and primary tooth root canal therapy (RCT) on cariously exposed vital pulps of primary molars. METHODS: A total of 291 molars were treated in 130 children--182 molars received FS and 109 received RCT by random selection. RESULTS: At 2-year reassessment, 116 molars (73 FS, 43 RCT) were available for clinical and radiographic examination. There was no clinical evidence of pathosis in 96% of FS and 98% of RCT molars. Two independent pediatric dentists evaluated periapical radiographs of the treated molars. Molars were classified into 1 of 4 outcomes: (1) N--normal treated molar, (2) H--nonpathologic radiographic change present, (3) P(O)--pathologic change present, follow-up in 6 months, and (4) P(X) -pathologic change present, extract immediately. Survival analysis was applied. A good level of agreement between raters was found for molars with outcome P(X) (kappa=0.745). Intrarater reliability was good for molars with outcome P(X) (kappa=0.710). Significantly greater numbers of FS than RCT molars were rated P(X) at the 2-year recall (chi2=5.8; P=.02). No significant difference in survival between the 2 types of vital pulp treatments was detected in log rank tests (P=.22). CONCLUSIONS: Outcomes for FS were poorer than RCT outcomes at 2 years; however, at 2 years, the survival rates were not statistically different. PMID- 12723833 TI - Assessing growth and development of the facial profile. AB - The study of growth and development of the facial profile is of interest to clinicians and researchers in the fields of pediatric dentistry, orthodontics, and craniofacial surgery, enabling diagnosis, planning, and evaluation of treatment. Until recently, craniofacial studies addressed facial growth, facial asymmetry, and gender differences by examining changes in size. However, size changes alone do not represent fully the complicated process of craniofacial growth which also involves changes in shape. The shape of the facial profile can now be quantified with Fourier analysis, contributing to a better understanding of growth. A combination of recently developed methods, such as 3-dimensional facial morphometry and Fourier analysis, should allow a more comprehensive knowledge of growth and development of the craniofacial structures, including the facial profile. This article examines various methods for assessing facial growth and development currently available with particular reference to the facial profile, and addresses the value of Fourier analysis in assessing shape changes. PMID- 12723834 TI - Interdental spacing and caries in the primary dentition. AB - PURPOSE: This study assessed the relationship between interdental spacing patterns and caries experience in the primary dentition. METHODS: Caries examinations were conducted amoung 356 children 4 to 6 years of age. At the time of the examinations, alginate impressions were obtained and poured in yellow stone. From the stone casts, each interdental area was categorized as: (1) space > 1 mm, (2) space < 1 mm, (3) no space, teeth in contact, or (4) no space, teeth overlapped. These categories were collapsed into presence or absence of space for each interdental site, and counted for each individual. Analyses assessed the relationships between interdental spacing and caries experience with separate analyses for anterior spacing, posterior spacing and total spacing. RESULTS: Children with more total interdental spaces had less decay experience and less untreated decay than children with fewer interdental spaces, and children with more molar spacing had less molar decay experience; however, these relationships were weak. Correlation analyses demonstrated significant relationships between number of decayed surfaces and total number of interdental spaces (r=-0.11, P=.04) and number of molar sites with interdental spaces (r=-0.13, P=.02). Multivariate analyses revealed the total number of interproximal spaces to be weakly associated with interproximal caries experience, but that fluoride exposure was a much stronger predictor. CONCLUSIONS: Absence of interdental spaces is weakly associated with greater decay experience in the primary dentition. PMID- 12723835 TI - Preschool caries as an indicator of future caries: a longitudinal study. AB - PURPOSE: This study was performed to compare the increment of carious surfaces per year in preschool-age children with early childhood caries (ECC), children with posterior caries only, and caries-free children after 7 to 10 years. METHODS: One hundred and fifty files of children were included in the study. The first examination referred to the ages of 3 to 5 years (T1), and follow-up visits took place after at least 7 years (T2). The number of carious surfaces was recorded. The study population was divided into 3 groups: (1) caries free children (CF), (2) children with ECC, and (3) children with posterior caries only (PC). There were 50 patients in each group. RESULTS: Children with ECC had 1.15+/ 0.97 new affected surfaces per year, while caries-free children had an increment per year of 0.41+/-0.60, and children with posterior caries only showed an increment per year of 0.74+/-0.64. A statistically significant difference was found between the ECC and caries-free groups, and between the ECC and posterior caries groups. The high increment in the ECC group is influenced by the high number of affected surfaces in the primary teeth, CONCLUSIONS: Children with ECC may have a high risk to develop future carious lesions compared with caries-free children. Children with posterior caries demonstrate less carious lesions by the age of 12 years, however, they resemble ECC children when they reach their mid teens. PMID- 12723836 TI - Fluoride varnish concentration gradient and its effect on enamel demineralization. AB - PURPOSE: Two of the 4 fluoride varnishes available on the American market today are sold in 10-mL tubes of 5% NaF varnish (Duraphat and Duraflor). Pilot studies have shown that a separation of contents within these tubes exists. The purpose of the current study was 4-fold: (1) to measure the fluoride concentration gradient in 10-mL tubes of fluoride varnish, based on resting position of the tube prior to use; (2) to compare and contrast fluoride concentration gradients of Duraphat, Duraflor, and CavityShield; (3) to compare this gradient to the ability to inhibit caries in an artificial caries environment; and finally, (4) to determine if quantitative light fluorescence (QLF) can detect differences in lesions developed when exposed to an artificial caries environment and fluoride varnish. METHODS: Human teeth specimens were subjected to a caries challenge and treated with a sample of fluoride varnish from 1 of 5 categories: Duraphat stored horizontally and vertically for 1 week, Duraflor stored horizontally and vertically for 1 week, or a CavityShield 0.4 mL "unidose." All specimens were then analyzed with the QLF system and with confocal microscopy. RESULTS: Results showed no significant fluoride ppm differences exist between groups (P=.29). It was shown that the order in which Duraflor varnish was dispensed from tubes significantly affected the fluoride concentration (P<.05). The order effect was not significant for Duraphat (P=.99). QLF data analysis showed there was no significant difference (P>.05) in the amount of remineralization obtained by using any varnish stored in any position. CONCLUSIONS: These results indicate that all 3 brands of fluoride varnish are able to remineralize incipient in vitro carious lesions, regardless of what part of the 10-mL tube the varnish is taken from. However, a fluoride concentration gradient exists in tubes of Duraflor. Also, QLF is able to detect demineralized and remineralized incipient lesions. PMID- 12723837 TI - The effect of acid primer or conventional acid etching on microleakage in a photoactivated sealant. AB - PURPOSE: The purpose of this study was to examine differences in microleakage in extracted human teeth when placing sealants using conventional acid etching or an acidic primer resin. METHODS: Three experimental groupings were used: group 1- conventional acid etching with placement of light-cured sealant; group 2- application of acidic primer resin (Prompt-L-Pop) and light curing, followed by sealant placement; group 3--similar to group 2, but acidic primer and sealant were photocured after placement of sealant. Teeth were thermocycled, stained, sectioned and examined for marginal microleakage. RESULTS: In group 1, 94% of the enamel-sealant interfaces were free of microleakage. For groups 2 and 3, only 28% showed no leakage, with most leakage occurring at both margin and base areas. Nonparametric data analysis indicated acid etching demonstrated significantly lower microleakage than either treatment using the acidic primer resin (P<.001) and that leakage scores in the acidic primer groups were identical (P=.4011). CONCLUSIONS: Use of this specific acidic resin primer in lieu of conventional acid etching (whether cured prior to or subsequent to sealant placement) demonstrated greater incidence of microleakage and would not be advocated over traditional etching procedures. PMID- 12723838 TI - Comparison of the def index with Nyvad's caries diagnostic criteria in 3- and 4 year-old Colombian children. AB - PURPOSE: The aims of this research were to determine the epidemiological profile of dental caries in 3- and 4-year-old preschool children living in Bogota, Colombia, and to compare two different caries indices--the standard def and Nyvad's new caries diagnostic criteria. METHODS: The children were screened by two calibrated examiners who first brushed the children's teeth and air dried them for 5 seconds before they were examined. The diagnostic criteria used were the standard def-t and def-s and the def-t and def-s of the new caries diagnostic system proposed by Nyvad. The chi-square test2 was used with a significance level of 5%. RESULTS: Prevalence of caries was 70% using the standard def-t criteria and 97% with the criteria proposed by Nyvad. The standard def-t and def-s were 3.3 and 5.7, respectively, and the def-t and def-s with the Nyvad citeria were 8.7 and 14.3, respectively. CONCLUSIONS: Prevalence of caries was high, indicating that the population studied had a high disease rate. The results obtained with the more detailed Nyvad new caries diagnostic criteria were higher than the ones obtained with the standard def-t index, both for teeth and surfaces. PMID- 12723839 TI - The effects of midazolam on pediatric patients with asthma. AB - PURPOSE: This study was performed to evaluate the safety and efficacy of midazolam in asthmatic patients undergoing dental treatment. METHODS: Twenty-four children, aged 19 to 65 months, with a diagnosis of mild to moderate asthma were given an oral dose of 0.5 mg/kg of midazolam. Oxygen saturation, respiratory rate, and pulse rate were monitored before, during, immediately after, and 30 minutes following dental treatment. The child's asthma score was also determined before and after treatment. The dental operator assessed the overall sedation outcome immediately after treatment. RESULTS: Twenty-three of the 24 subjects had asthma scores of "0" before and after treatment. During dental treatment, 2 patients had oxygen saturations of 94% at one point during treatment. However, oxygen saturation increased when the patient's head and neck were repositioned. Twenty-three of the 24 subjects had oxygen saturations above 95% at 30 minutes following treatment. Pulse rates and respiratory rates exhibited transitory increases, linked to when the child was stimulated. Statistical analysis was conducted from within subjects repeated measures via ANOVA and with a general linear model approach. No statistically significant differences occurred in oxygen saturation and respiratory rate. However, significant differences did occur in pulse rate between 5 and 10 minutes (mean difference=10+/-3.84) and between 10 and 15 minutes (mean difference=19+/-5.50), as expected. No statistically significant differences occurred in asthma score before and after treatment. Twelve subjects were assessed to have excellent behavior, 5 subjects were satisfactory, and 7 subjects were unsatisfactory. No treatment was aborted. CONCLUSIONS: With adherence to the AAPD's sedation guidelines, midazolam at a dose of 0.5 mg/kg is a safe and effective mean for sedation of patients with mild to moderate asthma. PMID- 12723840 TI - The detection of Porphyromonas gingivalis, Prevotella intermedia, and Actinobacillus actinomycetemcomitans in the supragingival plaque of children with and without caries. AB - PURPOSE: The present study was undertaken to determine the presence of 3 periodontopathic bacteria in the supragingival plaques of 3- to 16-year-old children with different oral health conditions. METHODS: DMFT and dft, PMA index (P=papillary gingivitis, M=marginal gingivitis, and A=attached gingivitis), OHI (oral hygiene index), and oral malodor of each subject were determined prior to the collection of supragingival plaques. Periodontopathic bacteria (P. gingivalis, P. intermedia, and A. actinomycetemcomitans) in supragingival plaques were detected using an immunoslot blot assay with monoclonal 3 periodontopathic bacteria in the 2 subject groups (children with and without caries). P. gingivalis-positive subjects, but not their P. intermedia or A. actinomycetemcomitans counterparts, were correlated to oral malodor. Oral malodor was also correlated to debris index, a component of OHI. RESULTS: The group with the higher OHI showed a higher prevalence of periodontopathic bacteria. For the 3 periodontopathic bacteria in the subjects tested, P. gingivalis-, P. intermedia-, and A. actinomycetemcomitans-positive plaques were not age related. CONCLUSIONS: The supragingival plaques in children can harbor periodontopathic bacteria such as P. gingivalis, P. intermedia, and A. actinomycetemcomitans. PMID- 12723841 TI - Condyloma acuminatum and human papilloma virus infection in the oral mucosa of children. AB - PURPOSE: The purpose of this study was to investigate the clinicopathological features of oral condylomas in children and condylomatous lesions of their mothers. Moreover, the authors sought to determine the mode of transmission of this disease and to find the genotype of human papilloma virus (HPV) in the children's oral condyloma. METHODS: Nine instances of oral condyloma acuminatum in children and lesions in their mothers were reviewed. Their HPV genotypes were evaluated by in situ hybridization (ISH). RESULTS: This study revealed that the lesions appeared during 3 years of age and the most common location was the hard and soft palate. Seven of the 9 mothers had experienced vulva and/or oral cavity condylomata during pregnancy. Social evaluation confirmed sexual abuse in 1 girl, and probable sexual abuse in another girl. The results of ISH demonstrated HPV 16/18 DNA being positive in 5 of the 9 cases, and HPV 6 and HPV 11, HPV 6 and HPV 16/18, HPV 6, and HPV 11 DNA being positive, respectively, in 1 case. HPV DNA types in mother-child pairs were not concordant. CONCLUSIONS: Oral condyloma acuminatum in children is probably induced by HPV 16/18. The mode of transmission by sexual abuse is the most likely route. Prenatal transmission of HPV to children is rare. This study provides further confirmation of possible different genotype and transmission in oral CA of children and adults. PMID- 12723842 TI - Prolonged bleeding following tongue piercing: a case report and review of complications. AB - The number of adolescents and young adults undergoing intraoral piercing is increasing in the United States. Numerous articles have documented complications following intraoral piercing. This article presents a case of prolonged bleeding and reviews other documented sequelae following intraoral piercing. The article may serve as a guide to dental professionals whose patients seek advice regarding these procedures. PMID- 12723843 TI - Repositioning an inverted maxillary central incisor using a combination of replantation and orthodontic movement: a clinical case report. AB - In some patients, the permanent central incisors fail to erupt and become impacted within the alveolus. In individuals with unfavorable impactions, surgical replantation may be considered to be a reasonable solution before orthodontic repositioning. In this clinical case report, a 9-year-old male patient presented with the permanent maxillary left central incisor in an inverted position high in the vestibule. He was treated with replantation and orthodontic repositioning with fixed appliances. Radiographic review 2 years post treatment showed an intact lamina dura consistent with a healthy tooth. It is concluded that orthodontic repositioning in combination with surgical replantation can be an optimal treatment choice for impacted teeth, assuming the anatomic location of the tooth is favorable. PMID- 12723844 TI - American Board of Pediatric Dentistry--the site visit checklist. PMID- 12723845 TI - Research on evidence-based practices: future directions in an era of recovery. AB - Many mental health systems are trying to promote the adoption of what has come to be known as evidence-based practices while incorporating a recovery vision into the services they provide. Unfortunately, much of the existing, published, research on evidence-based practices was conceived without an understanding of the recovery vision and/or implemented prior to the emergence of the recovery vision. As result, evidence-based practice research that has been published to date is deficient in speaking to a system being built on a recovery philosophy and mission; these deficiencies are detailed, and suggestions are advanced for new directions in evidence-based practice research. PMID- 12723847 TI - An expanded version of the Multnomah Community Ability Scale: anchors and interview probes for the assessment of adults with serious mental illness. AB - Objective clinical assessments are important in psychiatric settings to assess patients' functioning and the outcome of rehabilitation interventions. We developed anchors of the Multnomah Community Ability Scale (MCAS) and tested the inter-rater reliability of the expanded instrument. Twenty patients receiving psychiatric rehabilitation services participated in a structured interview and were rated by two raters. Intraclass correlation coefficients were .96 for the total and .87-.99 for the subscale scores. The expanded MCAS can serve as a reliable assessment tool. PMID- 12723848 TI - Poverty and mental health: a qualitative study of residential care facility tenants. AB - The impact of poverty on daily living was assessed for twenty-two mental health consumers using qualitative in-depth interviews. Consumers reported considerable difficulty meeting basic needs, such as clothing, shoes and personal care items. A majority reported running out of money before the end of each month. Respondents used strategies including careful budgeting, borrowing and employment to cope with poverty. Analysis indicated that the poverty experienced by respondents worked directly against participation in meaningful activities and their ability to build and sustain social relationships. Moreover, poverty directly contributed to stigma experienced by respondents. Findings suggest that a failure to recognize poverty as a key mental health policy issue will continue to constrain efforts to facilitate consumer empowerment and social integration. PMID- 12723849 TI - The emotional distress in a community after the terrorist attack on the World Trade Center. AB - OBJECTIVES: To examine psychological impact of the September 11th disaster on the immediate neighborhood of the New York World Trade Center. METHODS: 555 residents from the local Chinatown community participated in the study. They were surveyed retrospectively on their emotional-distress immediately after the tragedy and five months later. RESULTS: Prevalent anxiety was found in general community residents and additional depression in those who lost family members or friends. The mental health condition of the community improved tremendously five months later, with the initial 59% of general residents having 4 or more emotional symptoms dropping to 17%. However, more than half of the community residents had persistently shown one or more symptoms of emotional distress. Those who had lost a family member or friend in the disaster showed significantly higher distress, with 90% of them had four or more major psychiatric symptoms during the first few weeks right after the disaster, and the rate dropped to 35% five months later. Overall, those in their 40s and 50s seemed to have had relatively higher emotional distress than both younger and older groups. DISCUSSION: Methodological limitations were discussed concerning retrospective reporting and sample characteristics. PMID- 12723846 TI - Further validation of the Psycho-Social Well-Being Scale (PSWS) with community clients. AB - To monitor and evaluate the use of evidence-based approaches with mentally ill persons who abuse alcohol and other drugs, administrators, practitioners and evaluators will need to incorporate brief, reliable, and valid instruments into daily practice. The current two-part study provides further validation of the Psycho-Social Wellbeing Scale (PSWS), a multidisciplinary "debriefing tool" designed to capture and summarize data on clients' wellbeing from multiple sources in team-based community care. In the current investigation the PSWS was correlated with a number of valid instruments including the Brief Psychiatric Rating Scale, the Role Functioning Scale, the Social Functioning-12, the Alcohol Use Disorders Identification Test, the Alcohol Use and Drug Use Scales along with other important indices. Results show strong evidence of internal consistency, concurrent and discriminant validity with these criterion variables. Regression modeling demonstrates that the two main subscales (psychological and social wellbeing) add significantly to a model predicting substance abuse problems. PMID- 12723851 TI - Obstructive sleep apnea and dentists. PMID- 12723850 TI - A study of treatment outcomes from atypical antipsychotic medications in the Virginia public system of community care. AB - This retrospective study examines treatment outcomes associated with atypical antipsychotic medications in areas of 1) treatment response; 2) employment; 3) living arrangements; and 4) hospitalization rates. The study included a random sample of adults with serious mental illness, who participated in treatment for at least 36 months. Study results suggest that the utilization of these medications, despite higher costs, promote cost savings of public dollars through reduced hospitalization and enhanced functioning/outcomes. The study results proved to be very vital to the Virginia State Legislature and key system policy makers and funding authorities, as important actions were taken to enhance consumer access to these important medications and essential services. PMID- 12723852 TI - Making the case for insurance TMD reimbursement. PMID- 12723853 TI - Dr. Melis comments on Chakfa, et al.'s article in the October 2002 issue of CRANIO. PMID- 12723854 TI - Changes in head position due to occlusal supporting zone loss during clenching. AB - To establish the relationship between the condition of the stomatognathic system and body posture, changes in the head position during clenching were observed and investigated when the occlusal supporting zone was lost unilaterally and bilaterally. The results were as follows: 1. Regardless of the occlusal conditions, the head position was changed by clenching; 2. The occlusal conditions did not affect the changed distance of the head position; 3. The head position was changed forward and down by clenching regardless of the condition of the occlusal supporting zone. The head position changed more laterally to the opposite side of the lost occlusal supporting zone by clenching with the occlusal supporting zone lost unilaterally rather than bilaterally. Based on this study, it is suggested that unilateral loss of the occlusal supporting zone may cause the neck muscles to become inharmonious and thus affect body posture. PMID- 12723855 TI - Effects of laterotrusive occlusal scheme and body position on bilateral sternocleidomastoid EMG activity. AB - This study was conducted to determine the effects of laterotrusive occlusal scheme and body position on bilateral sternocleidomastoid electromyographic (EMG) activity. The study was performed on 20 healthy subjects with natural dentition and bilateral molar support. During laterotrusive occlusal excursion (working side), each individual had to present canine guidance on one side and group function on the opposite side, without balancing-side contacts. Integrated EMG (IEMG) recordings were performed by placing bipolar surface electrodes on the right and left sternocleidomastoid muscles. IEMG activity was recorded seated upright with the head unsupported and in the right and the left lateral decubitus body positions (head, neck and body horizontally aligned), under the following experimental conditions: 1. Maximal voluntary clenching in the intercuspal position; 2. Laterotrusive occlusal excursion with canine guidance; 3. Laterotrusive occlusal excursion with group function. Bilateral sternocleidomastoid IEMG activity with canine guidance or group function was significantly lower than the intercuspal position in both body positions. In the seated upright position, significantly lower activity was observed with canine guidance than in group function. In the lateral decubitus position activity was similar with both laterotrusive occlusal schemes and significantly higher than seated upright. Results suggest that both laterotrusive occlusal scheme and body position have significant influence on sternocleidomastoid IEMG activity. The clinical relevance of both factors is discussed. PMID- 12723856 TI - Mandible and maxilla bone mineral density and threshold analysis studies by pQCT in two edentulous women receiving pamidronate. AB - The mandible and maxilla of two edentulous women, treated during 25 weeks with oral pamidronate, were monitored by peripheral quantitative tomography (pQCT). Whole bone volumetric mineral density failed to show meaningful variations after treatment. However, an analysis of separated cortical and medullar areas disclosed focal bone loss at the right mandible cortex of patient #1 and at the left maxilla cortex of patient #2. These and other bone sub-regions were further studied by clustering the internal sites with a different degree of bone mineral density, resorting to the mineral threshold analysis provided by the system. Where bone loss was detected, it corresponded to increased loss of the most osteopenic sites, while medium and high-density portions tended to remain unchanged within the region. There were no significant variations in all other regions, or alternatively, minor losses at osteopenic sites were compensated by an increase at high-density portions. Hence, the pQCT system allowed monitoring volumetric bone mineral density at particular sites of interest, discriminating variations at portions with a dissimilar degree of bone volume. Further studies should confirm whether pamidronate exerts a protective effect on sub-regions with previous medium and high degrees of bone mineralization, as suggested by our present findings. PMID- 12723857 TI - Therapeutic response of benzodiazepine, orphenadrine citrate and occlusal splint association in TMD pain. AB - Loss of function, muscle inflammation, and pain are some of the signs and symptoms of temporomandibular dysfunction (TMD). Pharmacological strategies to minimize the clinical manifestation of these disorders often focus on blocking or inhibiting the pain-causing symptom. Resources such as muscle-relaxants, anxiety relief drugs, and splint therapy are often used to reduce muscular hyperactivity related to TMD muscle pain. This study compares the effect of a randomly ordered association of occlusal splint therapy (S), nonsteroid anti-inflammatory with a muscle-relaxant drug (orphenadrine citrate) (O), and an anxiety-relief drug (benzodiazepine) (B), to ease painful TMD muscle symptoms. Clinical and anamnestic analyses were recorded in accordance with the Helkimo TMD index and applied before and after treatments. Twenty-one group two Helkimo TMD adult female patients were treated, all of whom were subjected to the three random therapeutic associations proposed: SBO, BOS, and OSB. The same operator applied the three specific associations over a period of 21 days in the proposed sequence, seven days for each therapy. The results show that all the groups presented the best results in terms of relief from pain after the therapeutic association (28.5% showed a decrease and 47.6% showed an absence of symptoms). No significant difference was observed among association therapeutic protocols. PMID- 12723858 TI - Relationship between cervical lordosis and facial morphology in Caucasian women with a skeletal class II malocclusion: a cross-sectional study. AB - Several published studies show a relationship between craniofacial morphology and head posture. The aim of this study was to evaluate the association between cervical lordosis angle and mandibular length from lateral skull radiographs, and to investigate the relationship between mandibular retrusion and cervical lordosis angle decreasing. The sample comprised 70 Caucasian adult women, average 27.4, in skeletal class II and Angle class II. Lateral skull radiographs were obtained in natural head position (mirror position). Ten morphological variables were individuated on tracings. In order to assess errors due to landmark identification, double measurements were made in ten randomly selected radiographs and were compared with Dahlberg's formula. A Sperman's rank correlation test showed a negative correlation (P < 0.01) between cervical lordosis and mandibular length (compensatory curvature of the cervical spine) and positive correlation between anterior cranial base and maxillary length (P < 0.05). Based upon the cross-sectional method, no conclusion was possible about the mechanism concerning these results. Future longitudinal studies in growing patients should be directed to understanding the extent of environmental and genotype influences on cervical lordosis angle. PMID- 12723859 TI - Septoturbinal surgery in contact point headache syndrome: long-term results. AB - Facial pain syndrome secondary to sinonasal pathology is reported by the International Headache Society (IHS) classification (1988). It is underlined that a clear and proven nasal pathology with adequate painful stimuli must be present, i.e., acute sinusitis, vacuum sinus, or other unspecified pathologies. No clear role of septal abnormalities and turbinate hypertrophy has been attributed in the genesis of pain by the IHS classification. One of the most difficult problems in dealing with patients with sinonasal headaches is the definition of the primary cause of the pain. In our experience possible guidelines are history, endoscopic evaluation, diagnostic blocks, and computed tomography. The data reported here is from a long-term follow-up study of facial pain in a group of 34 patients with facial pain and nasal obstruction due to septoturbinal contact that did not respond to medical therapy. Patients, free from sinus disease or other causes of headache, were treated by septoplasty/rhinoseptoplasty,and middle turbinate electrocauterization. Pre- and postoperative patency was assessed by endoscopic evaluation and nasal resistance was assessed by anterior rhinomanometry. Patients were interviewed regarding pre- and post-operative intensity of pain (subjective pain was evaluated using the 0-10 Visual Analogue Scale (VAS) and frequency of the facial pain. The follow-up period ranged from 12 to 47 months (mean: 26.7 +/- 8.5 months). In 25% of the cases the pain relapsed post-operatively (from two days to one year); but in only three patients (8%) the relapses were persistent. Two out of three, however, reported a decreased VAS score after surgery. These results seem to indicate septoplasty and turbinate decongestion to be a fairly good surgical option in treating facial pain due to septoturbinal contact resistant to conservative nasal therapy. PMID- 12723860 TI - Reduction in sick leave and costs to society of patients with Meniere's disease after treatment of temporomandibular and cervical spine disorders: a controlled six-year cost-benefit study. AB - This study compares the frequency of sick leave between the three-year period after and the three-year period before coordinated treatment of temporomandibular and cervical spine disorders in 24 patients (ten males and 14 females) diagnosed with Meniere's disease. The frequency of sick leave for the patients was also compared with the frequency of sick leave in a control group from the population. A cost-benefit analysis was made regarding the costs to society of sick leave related to the treatment costs of the patients. In a previous study the same patients were treated for their severe signs and symptoms of temporomandibular and cervical spine disorders, and they reported a substantial reduction in their vertigo, non-whirling dizziness, tinnitus, feeling of fullness in the ear, pain in the face and jaws, pain in the neck and shoulders, and headache. The number of days of sick leave and the year the patient began to receive disability pension due to the symptoms of Meniere's disease were obtained from the National Health Insurance Service in Sweden. Two of the patients received disability pension benefits due to Meniere's disease 17 years prior to their normal retirement pension. A third patient received disability pension for another reason and two were receiving a retirement pension. Data on the remaining 19 patients showed a considerable reduction in number of days of sick leave during the three-year period after coordinated treatment (270 days) compared with the three-year period before the treatment (1,536 days). The control subjects used a total of 14 days sick leave for the same symptoms during the same six-year period. Vertigo (nine days) was the dominant cause followed by pain in the neck and shoulders, and headache. The reduction in sick leave for the 19 nonretired patients and the treatment costs for the 24 patients can be used for a simple cost-benefit calculation for the subgroup of nonretired patients. During the first three years after treatment the reduction in sick leave was on average 66.6 days for each of the 19 nonretired patients. Within the limits of this study, it can be concluded that the costs to society for sick leave and disability pension due to Meniere's disease are substantial. A coordinated treatment of temporomandibular and cervical spine disorders appears to substantially reduce these costs. PMID- 12723861 TI - Prevalence of bruxism awareness in a Sardinian population. AB - 1014 subjects on the island of Sardinia (Italy) were interviewed regarding the habit of clenching and grinding their teeth. They had to specify if this activity occurred during the day, during the night, or both. Other information recorded was their age, gender, marital status, and occupation. Overall prevalence of bruxism was 27.2% (276 subjects). No association was found between bruxism and age, gender and job. Even differentiating diurnal, nocturnal, diurnal and nocturnal bruxism, associations were non-significant. Marital status seems to make some difference: divorced people reported higher parafunctional activity compared to widows and widowers who reported the least. Although awareness of bruxism is not a precise measure of parafunction, based on the results we cannot support the role of stress on bruxism etiology. PMID- 12723862 TI - Orofacial pain of cardiac origin: a case report. AB - It is the responsibility of the dentist to make the differential diagnosis of pain in the region of the oral cavity. Cardiac pain most commonly radiates to the left arm, shoulder, neck and face. In rare instances the pain may present as dental pain. In this case report, the patient presented with complaints of intense bilateral pain in the jaws which was diagnosed as pain of cardiac origin. PMID- 12723863 TI - Treatment efficacy of noncontingent reinforcement during brief and extended application. AB - We evaluated the long-term therapeutic effects of noncontingent reinforcement (NCR). In Experiment 1, NCR effects were examined with 2 participants' arbitrary responses; in Experiment 2, NCR was used as treatment with 3 participants whose self-injurious behavior (SIB) was maintained by automatic reinforcement. In both experiments, NCR consisted of continuous access to a highly preferred leisure item and was implemented initially during 10-min and later during 120-min sessions. Varied reinforcers (leisure items) were subsequently introduced during 120-min sessions to determine if treatment effects might be extended. Finally (Experiment 2 only), NCR was implemented throughout the day in participants' homes. Results of Experiments 1 and 2 showed that reinforcers obtained through object manipulation can compete with those obtained automatically by engaging in SIB during brief NCR sessions. However, data from the 120-min sessions indicated that satiation to a specific leisure item might occur over periods of time more typical of those during which treatment would be implemented. Access to a variety of highly preferred leisure items extended the effectiveness of NCR for some individuals. When NCR was implemented throughout the day (Experiment 2), therapeutic effects were shown to be maintained for up to 1 year. PMID- 12723864 TI - Analysis of precurrent skills in solving mathematics story problems. AB - We conducted an analysis of precurrent skills (responses that increase the effectiveness of a subsequent or "current" behavior in obtaining a reinforcer) to facilitate the solution of arithmetic word (story) problems. Two students with developmental disabilities were taught four precurrent responses (identifying the initial value, change value, operation, and resulting value) in a sequential manner. Results of a multiple baseline design across behaviors showed that the teaching procedures were effective in increasing correct performance of each of the precurrent behaviors with untaught problems during probes and that once the precurrent behaviors were established, the number of correct problem solutions increased. PMID- 12723865 TI - Progressing from programmatic to discovery research: a case example with the overjustification effect. AB - Scientific research progresses along planned (programmatic research) and unplanned (discovery research) paths. In the current investigation, we attempted to conduct a single-case evaluation of the overjustification effect (i.e., programmatic research). Results of the initial analysis were contrary to the overjustification hypothesis in that removal of the reward contingency produced an increase in responding. Based on this unexpected finding, we conducted subsequent analyses to further evaluate the mechanisms underlying these results (i.e., discovery research). Results of the additional analyses suggested that the reward contingency functioned as punishment (because the participant preferred the task to the rewards) and that withdrawal of the contingency produced punishment contrast. PMID- 12723866 TI - Response-restriction analysis: I. Assessment of activity preferences. AB - We used procedures based on response-restriction (RR) analysis to assess vocational and leisure activity preferences for 3 adults with developmental disabilities. To increase the efficiency of the analysis relative to that reported in previous research, we used criteria that allowed activities to be restricted at the earliest point at which a preference could be determined. Results obtained across two consecutive RR assessments showed some variability in overall preference rankings but a high degree of consistency for highly ranked items. Finally, we compared results of the RR assessment with those of an extended free-operant assessment and found that the RR assessment yielded (a) more differentiated patterns of preference and (b) more complete information about engagement with all of the target activities. PMID- 12723867 TI - Response-restriction analysis: II. Alteration of activity preferences. AB - We used response-restriction (RR) assessments to identify the preferences of 7 individuals with mental retardation for a variety of vocational and leisure activities. We subsequently increased their engagement in nonpreferred activities using several procedures: response restriction per se versus a Premack-type contingency (Study 1), supplemental reinforcement for engagement in target activities (Study 2), and noncontingent pairing of reinforcers with nonpreferred activities (Study 3). Results indicated that preferences are not immutable and can be altered through a variety of relatively benign interventions and that the results of RR assessments may be helpful in determining which types of procedures may be most effective on an individual basis. PMID- 12723868 TI - Pyramidal training for families of children with problem behavior. AB - The pyramidal training model was extended to multiple family members of children with behavior disorders. Three primary caregivers were taught to implement individualized treatments for problem behavior. They were then taught how to use various instructional strategies (e.g., prompting, feedback) to teach 2 other family members to implement the treatment. Results showed that pyramidal training was effective in increasing caregiver implementation of treatments across three families. PMID- 12723869 TI - Acquisition of cup drinking using previously refused foods as positive and negative reinforcement. AB - We used previously refused foods as positive and negative reinforcement in the acquisition of cup drinking. Cup drinking increased with positive and negative reinforcement, both alone and in combination (without escape extinction), indicating that treatment of food refusal can establish some foods as appetitive stimuli whereas others remain aversive. PMID- 12723870 TI - Recombinative generalization of within-syllable units in nonreading adults with mental retardation. AB - Two adults with mental retardation demonstrated the recombination of within syllable units (onsets and rimes) using a spoken-to-printed-word matching-to sample (MTS) procedure. Further testing with 1 participant showed comprehension of the printed words. Printed-word naming was minimal before, but greater after, comprehension tests. The findings suggest that these procedures hold promise for further basic and applied analyses of word-attack skills. PMID- 12723871 TI - Covariation between bizarre and nonbizarre speech as a function of the content of verbal attention. AB - A functional analysis suggested that the bizarre speech of an individual with developmental disabilities was maintained by attention. The content of verbal attention was manipulated in two subsequent analyses and revealed that (a) bizarre speech was more frequent when attention was related to the participant's bizarre speech and (b) the participant's statements tended to reflect the content of the therapist's attention, whether bizarre or nonbizarre. PMID- 12723872 TI - Use of the high-probability instructional sequence and escape extinction in a child with food refusal. AB - We used the high-probability (high-p) instructional sequence with and without escape extinction in the treatment of food refusal. Acceptance increased and refusal decreased only with the introduction of escape extinction. These results raise important questions about the high-p sequence in the treatment of food refusal. PMID- 12723873 TI - Treatment of vocal tics in children with Tourette syndrome: investigating the efficacy of habit reversal. AB - Habit reversal was used to treat vocal tics in 5 children with Tourette syndrome. Vocal tics were reduced in 4 of the 5 children, the untreated motor tics did not increase, and treatment was acceptable to the children's parents. PMID- 12723874 TI - Citation trends of applied journals in behavioral psychology 1981-2000. AB - One variable with which to evaluate scientific journals is how often their articles are cited in the literature. Such data are amenable to longitudinal analysis and can be used as a measure of a journal's impact on research within a discipline. We evaluated multiple citation measures for a number of applied journals in behavioral psychology from 1981 to 2000. The results indicate a relatively consistent impact across these journals, with some evidence of growth. PMID- 12723875 TI - Collateral effects of response blocking during the treatment of stereotypic behavior. AB - The collateral effects of response blocking were evaluated while treating stereotypic behavior in a woman diagnosed with autism. Blocking stereotypic behavior (head and tooth capping) was associated with decreases in leisure-item interaction and increases in another stereotypic response (hand wringing). Results suggested that the reduction in item interaction was due to adventitious punishment. Prompts to access an alternative source of reinforcement attenuated the side effects somewhat, but results suggested that the undesirable effects of response blocking may be fairly durable. PMID- 12723877 TI - Brief training to promote the use of less intrusive prompts by nursing assistants in a dementia care unit. AB - We evaluated the efficacy of a brief staff-training procedure to increase the use of graduated prompting by 2 certified nursing assistants (CNAs) while they helped to dress 3 persons with dementia in a seven-bed dementia care unit. The multiple baseline design across participants showed that CNAs dressed residents with minimal resident involvement during baseline observations. Following brief in service training, CNAs provided graduated prompts and praise appropriately, suggesting that CNAs can promote active involvement in personal care routines by older adults with dementia. PMID- 12723876 TI - An evaluation of a brief functional analysis format within a vocational setting. AB - We conducted and compared both brief and extended functional analyses of disruptive behaviors for 3 individuals with developmental disabilities who attended a vocational training program. Results demonstrated that the brief assessment identified the function of 2 of the 3 participants' disruptive behavior compared to the extended assessment. PMID- 12723878 TI - Effects of teacher-directed versus student-directed instruction on self management of young children with disabilities. AB - In this study, students worked independently by setting goals, selecting assignments, and recording and evaluating their results after receiving one of two different types of self-management training. During teacher-directed training, the teacher set goals, assigned work, and recorded and evaluated results for students. During student-directed training, students performed those tasks themselves. The results indicated that students engaged in the self management behaviors more frequently during independent work following student directed instruction than following teacher-directed instruction. PMID- 12723880 TI - Seasonal affective disorder: an overview. AB - Seasonal Affective Disorder (SAD) is a condition of regularly occurring depressions in winter with a remission the following spring or summer. In addition to depressed mood, the patients tend to experience increased appetite and an increased duration of sleep during the winter. SAD is a relatively common condition, affecting 1-3% of adults in temperate climates, and it is more prevalent in women. The pathological mechanisms underlying SAD are incompletely understood. Certain neurotransmitters have been implicated; a dysfunction in the serotonin system in particular has been demonstrated by a variety of approaches. The role of circadian rhythms in SAD needs to be clarified. The phase-delay hypothesis holds that SAD patients' circadian rhythms are delayed relative to the sleep/wake or rest/activity cycle. This hypothesis predicts that the symptoms of SAD will improve if the circadian rhythms can be phase-advanced. There is some experimental support for this. SAD can be treated successfully with light therapy. In classical light therapy, the SAD sufferer sits in front of a light box, exposed to 2000-10,000 lux for 30-120 min daily during the winter. Other forms of light treatments, pharmacotherapy, and other therapies are currently being tested for SAD. PMID- 12723879 TI - Electrophysiology of the circadian pacemaker in mammals. AB - The neurons of the mammalian suprachiasmatic nuclei (SCN) control circadian rhythms in molecular, physiological, endocrine, and behavioral functions. In the SCN, circadian rhythms are generated at the level of individual neurons. The last decade has provided a wealth of information on the genetic basis for circadian rhythm generation. In comparison, a modest but growing number of studies have investigated how the molecular rhythm is translated into neuronal function. Neuronal attributes have been measured at the cellular and tissue level with a variety of electrophysiological techniques. We have summarized electrophysiological research on neurons that constitute the SCN in an attempt to provide a comprehensive view on the current state of the art. PMID- 12723881 TI - The precision of circadian clocks: assessment and analysis in Syrian hamsters. AB - Locomotor activity recordings of Syrian hamsters were systematically analyzed to estimate the precision of the overt circadian activity rhythm in constant darkness. Phase variation, i.e., the standard deviation of phase markers around the regression line, varied with the definition of phase. Smallest phase variation was found in the onset of wheel running activity defined by 1h running means of the raw data. Both lower and higher degrees of smoothing lead to decreased precision measured in the overt rhythm. With passive infrared recordings, the midpoint of activity defined by 3h running means was the least variable. This demonstrates that the choice of phase marker should vary between recording methods. Phase variation decreased with increasing activity and was larger in females than in males. By calculating the average cycle variation and serial covariance of consecutive cycles, we estimated the contribution of 'clock' and 'non-clock' related processes to the overt rhythm variability. Variance in precision between phase markers could be shown to be attributable mainly to nonclock processes. Variance in pacemaker cycle length appeared reduced in wheel running activity records compared with passive infrared sensing records, suggesting feedback from running activity onto pacemaker function. PMID- 12723882 TI - Effect of light intensity on the phase and period response curves in the nocturnal field mouse Mus booduga. AB - The effect of light intensity on the phase response curve (PRC) and the period response curve (tauRC) of the nocturnal field mouse Mus booduga was studied. PRCs and tauRCs were constructed by exposing animals free-running in constant darkness (DD), to fluorescent light pulses (LPs) of 100 lux and 1000 lux intensities for 15 min duration. The waveform of the PRCs and tauRCs evoked by high light intensity (1000 lux) stimuli was significantly different compared to those constructed using low light intensity (100 lux). Moreover, a weak but significant correlation was observed between phase shifts and period changes when light stimuli of 1000 lux intensity were used; however, the phase shifts and period changes in the 100lux PRC and tauRC were not correlated. This suggests that the intensity of light stimuli affects both phase and period responses in the locomotor activity rhythm of the nocturnal field mouse M. booduga. These results indicate that complex mechanisms are involved in entrainment of circadian clocks, even in nocturnal rodents, in which PRC, tauRC, and dose responses play a significant role. PMID- 12723883 TI - Plasticity of hamster circadian entrainment patterns depends on light intensity. AB - The multiple oscillatory basis of the mammalian circadian pacemaker is adduced by, among other phenomena, the occurrence of split locomotor activity rhythms in rodents after prolonged exposure to constant light. More recently, split rhythms entrained to a 24h light:dark:light:dark cycle have been documented following scheduled access of hamsters to a novel running wheel or by photoperiod manipulations alone. Because the incidence of constant light-induced splitting depends on light intensity, the role of this variable was assessed in this new splitting paradigm. Male Syrian hamsters, entrained to a 14h light:10h dark cycle, were transferred to individual running wheel cages 7h after light onset. Transfer coincided with the beginning of the scotophase of a new photocycle alternating between 5h of relative dark and 7h of light. For four weeks bright photophases (approximately 350 lux) were alternated with either dim (< 0.1 lux) or completely dark (0 lux) scotophases. An additional group received moderate intensity photophases (approximately 45 lux) paired with dim scotophase illumination. For an additional four weeks, all hamsters were exposed to the same bright:dim light:dark cycle. Dim light in the scotophase significantly increased the incidence of split activity rhythms relative to that observed with completely dark scotophases. Overall wheel-running rates and activity induced by a cage change were also increased in dim light-exposed animals. Group differences largely disappeared four weeks later when hamsters previously maintained in completely dark scotophases were exposed to dim scotophases. Photophase light intensity did not affect the overall incidence of splitting, but influenced the timing of activity in the afternoon scotophase. The effects of dim illumination may be mediated in part via enhanced locomotor responses to transfer to a new cage or by changes in coupling interactions between component oscillators. PMID- 12723884 TI - The development of new purification methods to assess the circadian rhythm of body temperature in Mongolian gerbils. AB - Six Mongolian gerbils were studied for 8-10d while housed in separate cages in a 12:12h light-dark (L-D) cycle (lights on at 07:00h). Recordings of body temperature, heart rate, and spontaneous activity were made throughout. The temperature and heart rate rhythms were "purified" to take into account the effects of activity, and then the rhythm of temperature was further purified to take into account other masking influences ("non-activity masking effects" or NAME,). The methods employed in the purification processes involved linear regression analysis or analysis of covariance, the latter using functions of activity and NAME as covariates. From these methods, it was possible to obtain not only an estimate of the endogenous component of the temperature rhythm but also a measure of circadian changes in the sensitivity of temperature to masking effects. Even though all purification methods removed many of the effects of spontaneous activity from the temperature record, there remained temperature fluctuations at the L-D and D-L transitions that appeared to be independent of activity. The NAME was of only very marginal value in the purification process. Comparison of the purification methods indicated that the linear methods were inferior (both from a biological viewpoint and when the results were compared mathematically) to those that allowed the rate of rise of temperature due to increasing amounts of activity to become progressively less. The sensitivity of temperature and heart rate to the masking effects of activity showed a circadian rhythm, with sensitivities in the resting phase being greater than those in the active phase. These findings are compatible with the view that thermoregulatory reflexes are induced by spontaneous activity of sufficient amount, and that there is a circadian rhythm in the body temperature at which these reflexes are initiated and in their effectiveness. PMID- 12723885 TI - Daily changes of GABA and taurine concentrations in various hypothalamic areas are affected by chronic hyperprolactinemia. AB - This study was designed to characterize, in anterior, mediobasal, and posterior hypothalamic and median eminence, the 24h changes of gamma aminobutyric acid (GABA) and taurine (TAU) contents in adult male rats and to analyze whether chronic hyperprolactinemia may affect these patterns. Rats were turned hyperprolactinemic by a pituitary graft. Plasma prolactin (PRL) levels increased after pituitary grafting at all time points examined. A disruption of the circadian rhythm was observed in pituitary-grafted rats, whereas GABA and TAU content followed daily rhythms in all areas studied in controls. In the mediobasal hypothalamus, two peaks for each amino acid were found at midnight and midday. In the anterior hypothalamus, GABA and TAU showed only one peak of concentration at midnight. In the posterior hypothalamus, the values of both GABA and TAU were higher during the light as compared to the dark phase of the photoperiod. In the median eminence GABA content peaked at 20:00h, the time when TAU exhibited the lowest values. Hyperprolactinemia abolished the 24h changes of GABA in the mediobasal hypothalamus and reduced its content as compared to controls. Hyperprolactinemia advanced the diurnal peak of TAU to 12:00h in the mediobasal hypothalamus and did not modify the 24:00h peak. In the anterior hypothalamus, hyperprolactinemia increased GABA and TAU contents during the light phase while it decreased them during the dark phase of the photoperiod. In the posterior hypothalamus hyperprolactinemia did not modify GABA or TAU patterns as compared to controls. In the median eminence hyperprolactinemia increased the 20:00h peak of GABA and shift advanced the decrease in TAU content at 20:00h and its maximum at 24:00h as compared to controls. These data show that GABA and TAU content exhibit specific daily patterns in each hypothalamic region studied. PRL differentially affects the daily pattern of these amino acids in each hypothalamic region analyzed. PMID- 12723886 TI - Effects of diazepam and its metabolites on nocturnal melatonin secretion in the rat pineal and Harderian glands. A comparative in vivo and in vitro study. AB - We investigated the effects of diazepam (DZP) and its three metabolites: nordiazepam (NZP), oxazepam (OZP), and temazepam (TZP) on pineal gland nocturnal melatonin secretion. We looked at the effects of benzodiazepines on pineal gland melatonin secretion both in vitro (using organ perifusion) and in vivo in male Wistar rats sacrificed in the middle of the dark phase. We also examined the effects of these benzodiazepines on in vivo melatonin secretion in the Harderian glands. Neither DZP (10(-5)-10(-6)M) nor its metabolites (10(-4)-10(-5)M) affected melatonin secretion by perifused rat pineal glands in vitro. In contrast, a 10(-4)M suprapharmacological concentration of DZP increased melatonin secretion of perifused pineal glands by 70%. In vivo, a single acute subcutaneous administration of DZP (3 mg/kg body weight) significantly affected pineal melatonin synthesis and plasma melatonin levels, while administration of the metabolites under the same conditions did not. DZP reduced pineal melatonin content (-40%), N-acetyltransferase activity (-70%), and plasma melatonin levels (-40%), but had no affects on pineal hydroxyindole-O-methyltransferase activity. Neither DZP nor its metabolites affected Harderian gland melatonin content. Our results indicate that the in vivo inhibitory effect of DZP on melatonin synthesis is not due to the metabolism of DZP. The results also show that the control of melatonin production in the Harderian glands differ from that observed in the pineal gland. PMID- 12723887 TI - Circadian response of annual weeds in a natural setting to high and low application rates of four herbicides with different modes of actions. AB - Four herbicides [glyphosate (GLYT), an amino acid synthesis inhibitor; glufosinate (GLUF), a glutamine synthetase inhibitor; fomesafen (FOME), a protoporphyrinogen oxidase inhibitor, and chlorimuron ethyl (CLIM), an acetolactate synthase inhibitor] were used to examine the influence of time of day of application on the control of a variety of annual broadleaf weeds in field studies conducted in Minnesota (five studies on GLYT and GLUF, three studies on FOME and CLIM). All herbicides were applied with an adjuvant at recommended high and low (half or quarter strength) rates every 3h between 06:00 and 24:00h local time. Visual ratings of percent weed control evaluated at 14d were analyzed by herbicide and application rate for each study and across studies for time-of-day effect by analysis of variance (ANOVA) and single cosinor. A circadian response to each herbicide was found, with greatest weed control observed between 09:00 and 18:00h. Increasing the herbicide application rate did not overcome the time of-day effect (ANOVA: p < or = 0.008 for time-of-day effect for each herbicide and application rate). The least-squares fit of a 24h cosine was significant (p < or = 0.001) for each herbicide and application rate, with double amplitudes of 18 82% (units = % visual control) and estimated peaks (acrophases) near midday between 12:40 and 13:35h. Analysis of residuals obtained from multiple regression that included weed height, herbicide rate, temperature, and relative humidity as independent factors also found a significant time-effect by both ANOVA and cosinor for each herbicide and rate, with acrophases advancing significantly by 3 to 7h for GLYT and GLUF, but not for FOME or CLIM. These results suggest that the four herbicides, while belonging to different families with different modes of action, may reveal different peak times of efficacy when adjusting for environmental factors. Nonetheless, each displays similar circadian patterns when influenced by these factors under natural seasonal field conditions. The within day rhythmic differences found in weed control are large enough to warrant consideration of the practical financial and environmental importance of the time of-day that these and other herbicides are applied. PMID- 12723888 TI - Circadian rhythms in serum bone markers and their relation to the effect of etidronate in rats. AB - Circadian rhythmicity is an essential feature of bone metabolism. The present study was undertaken to (Aoshima et al., 1998) determine the changes in bone resorption and formation in rats over 24h, (Black et al., 1999) evaluate the effect of the consecutive administration of etidronate on circadian rhythms of serum bone markers, and (Blumsohn et al., 1994) determine whether the effect of etidronate on bone metabolism is circadian time-dependent. One hundred twenty male Wistar rats, which had been adapted to a 12/12h light/dark cycle, were injected subcutaneously once daily with either 0.5 mgP/kg etidronate or 0.9% NaCl (control group) at 0090, 1300, 1700, 2100, 0100, or 0500h for 10d. Serum was collected and tibiae were dissected 24h after the last injection. Serum pyridinoline (Pyd), tartrate-resistant acid phosphatase (TRAP), osteocalcin (OC), alkaline phosphatase (ALP), calcium (Ca), phosphorus (Pi), calcitonin (CT), and parathyroid hormone (PTH) were determined. Bone mineral density (BMD) in the proximal tibia, and the rate of formation of longitudinal trabecular bone over the past 48h were also determined using a chronological labeling method with NTA Pb. The results showed characteristic circadian rhythms in serum bone markers in rats, with peaks in both bone resorption and bone formation during the animals' rest span. The administration of etidronate at the different times of the day decreased the level of bone-resorption markers (Pyd and TRAP) without affecting the circadian patterns of markers of bone formation (OC and ALP). However, the magnitude of the decrease due to etidronate was not uniform throughout the day, and was greatest during the daytime. Etidronate increased the BMD in the tibial metaphysis in all of the time-treatment groups, but the magnitude of the increase did not vary with the time of etidronate administration. The present data provide a physiological basis for future studies on bone metabolism and may be important in the design of future experiments and in the interpretation of experimental data. PMID- 12723889 TI - Comparison of circadian rhythm characteristics of blood pressure and heart rate in patients before and after elective coronary artery bypass surgery. AB - Coronary artery bypass grafting surgery (CABGS) is done to reperfuse the ischemic myocardium of coronary disease patients. This study was designed to analyze the circadian rhythm characteristics of blood pressure (BP) and heart rate (HR) of patients before and after CABGS. Fifty-one patients undergoing elective CABGS were studied; 21 patients received one, 12 two and 18 three or more grafts. BP was monitored for 24h before and after CABGS while patients were recumbent in the hospital. Systolic (S) and diastolic (D) BP and HR were assessed every 30 min. Of the 51 patients, 37 (73%) had nondipper 24h BP patterns (nocturnal decline in BP < 10% of daytime mean level) in the preoperative baseline assessment. The peak and MESOR (rhythm-adjusted 24h mean) values of the circadian rhythm in SBP, DBP, and pulse pressure (PP) significantly declined following surgery, while HR and rate-pressure product (RPP = SBP x HR) markedly increased. The double amplitude (peak-to-trough variation) of the circadian rhythm in SBP and DBP was significantly reduced postoperatively, and that of the rhythm in HR and RPP significantly increased. The slopes of the morning rise and evening dip in the 24h SBP profile were reduced significantly after bypass grafting. The corresponding slopes of the HR profile, in contrast, were markedly increased. PMID- 12723890 TI - Twelve-hour night shifts of healthcare workers: a risk to the patients? AB - We assessed the impact of 12h fixed night shift (19:00-07:00h) work, followed by 36h of off-time, on the sleep-wake cycle, sleep duration, self-perceived sleep quality, and work-time alertness on a group composed of 5 registered and 15 practical nurses. Wrist actigraphy (Ambulatory Monitoring, Inc.), with data analysis by the Cole-Kripke algorithm, was applied to determine sleep/wake episodes and their duration. The sleep episodes were divided into six categories: sleep during the night shift (x = 208.6; SD +/- 90.6 mins), sleep after the night shift (x = 138.7; SD +/- 79.6 min), sleep during the first night after the night work (x = 318.5; SD +/- 134.6 min), sleep before the night work (x = 104.3; SD +/ 44.1 min), diurnal sleep during the rest day (x = 70.5; SD +/- 43.0 min), and nocturnal sleep during the rest day (x = 310.4; SD +/- 188.9mins). A significant difference (p < .0001; T-test for dependent samples) was detected between the perceived quality of sleep of the three diurnal sleep categories compared to the three nocturnal sleep categories. Even thought the nurses slept (napped) during the night shift, their self-perceived alertness systematically decreased during it. Statistically significant differences were documented by one-way ANOVA (F = 40.534 p < .0001) among the alertness measurements done during the night shift. In particular, there was significant difference in the level of perceived alertness (p < .0001) between the 7th and 10th of the 12h night shift. These findings of decreased alertness during the terminal hours of the night shift are of concern, since they suggest risk of comprised patient care. PMID- 12723891 TI - Fluoride-induced apoptosis in human epithelial lung cells (A549 cells): role of different G protein-linked signal systems. AB - In the present study, possible mechanisms involved in fluoride-induced apoptosis in a human epithelial lung cell line (A549) were examined. Sodium fluoride (NaF) induced apoptosis in the A549 cells, with a maximum at 5-7.5 mM after 20 hours of exposure. The number of cells with plasma membrane damage (PI-positive cells) increased moderately up to 5 mM, but markedly at 7.5 mM. Deferoxamine (an Al3+ chelator) almost completely prevented these NaF-induced responses, which may suggest a role for G protein activation. The apoptotic effect was partially reduced by the PKA inhibitor H89. NaF induced a weak but sustained increase in PKC activity, whereas the PKC activator TPA induced a transient effect. TPA, which enhanced the NaF-induced PKC activity, was not apoptotic when added alone, but facilitated the NaF-induced apoptosis and the increase in PI-positive cells. PKC downregulation induced by TPA pretreatment almost completely prevented the NaF-induced apoptosis and the increase in PI-positive cells. Pretreatment with the PKC inhibitor GF109203X, which abolished the PKC activity after 3 hours, enhanced the NaF-induced apoptosis. KN93 (a CaM kinase II inhibitor) and W7 (a calmodulin inhibitor) seem to reduce the apoptotic effect of NaF, whereas BAPTA AM (a Ca2+ chelator) was without effect. The tyrosine kinase inhibitor genistein also markedly reduced the NaF-induced apoptosis, whereas the PI-3 kinase inhibitor wortmannin augmented the response. In conclusion, the present results suggest that NaF induces an apoptotic effect and an increase in PI-positive A549 cells via similar mechanisms, involving PKC, PKA, tyrosine kinase and Ca2+-linked enzymes, whereas PI-3 kinase seems to exert a counteracting effect. PMID- 12723892 TI - Correlation of lead and cadmium in human seminal plasma with seminal vesicle and prostatic markers. AB - The objective of this study was to investigate the correlation between lead and cadmium with seminal vesicle and prostatic markers. Semen samples categorized into fertile and infertile were evaluated for the presence of lead and cadmium and biochemical markers in the seminal plasma. Associations between lead and fructose, acid phosphatase and gamma-glutamyl transpeptidase (gamma-GT) were observed. However, no such relationships were noticed for cadmium. It is concluded that lead may be one of the pollutants indirectly affecting semen quality by altering the functions of accessory sex glands. PMID- 12723893 TI - Protective role of ascorbic acid and alpha-tocopherol on arsenic-induced microsomal dysfunctions. AB - Arsenic, a naturally occurring element, is present in food, soil, air and water. All human populations are exposed to arsenic and its compounds through occupational or environmental processes. Since arsenic compounds have been shown to exert their toxicity chiefly by generating reactive oxygen species, we have evaluated the effect of ascorbic acid and alpha-tocopherol on oxidative damage, antioxidant status and on xenobiotic metabolizing systems in arsenic-exposed rat liver and kidney microsomes. Arsenic exposure increases oxidative damage to lipids and proteins and decreases the levels of antioxidants and the activities of xenobiotic metabolizing enzymes. Coadministration of ascorbic acid and alpha tocopherol to arsenic-exposed rats resulted in a reduction in the levels of lipid peroxidation, protein carbonyls and hydrogen peroxide and an elevation in the levels of reduced glutathione, ascorbic acid and alpha-tocopherol. Ascorbic acid and alpha-tocopherol treatment decreases the activity of haem oxygenase, whereas it increases the levels/ activity of cytochrome P450, cytochrome b5 and NADPH cytochrome P450 reductase in arsenic-intoxicated rats. The results of this study provide evidence that ascorbic acid and alpha-tocopherol supplementation can improve the arsenic-induced altered microsomal functions in liver and kidney. PMID- 12723894 TI - A comparative study on the gastroduodenal tolerance of different antianaemic preparations. AB - The most significant adverse effect of repeated oral administration of iron containing antianaemic preparations is the gastroduodenal toxicity, attributable to a direct toxic effect of iron on the glandular epithelium. To assess gastroduodenal mucosal damage and the potential protective effect of different antianaemic preparations, a study was carried out to compare the gastroduodenal toxicity caused by three different types of antianaemic drugs in normal and anaemic rats administered at repeated therapeutic doses. Histological damage to the gastroduodenal mucosa was evaluated using light and electron microscopy. In both normal and anaemic rats, pathological changes were less marked in animals treated with ferrimannitol-ovoalbumin (TM/FMOA) than in those treated with iron protein succinylate or ferrous sulphate. Electron microscopic studies of duodenal mucosa in normal rats treated with iron protein succinylate and ferrous sulphate confirmed a severe ultrastructural alteration, whereas no changes were detected in animals treated with TM/FMOA. In anaemic rats, slight duodenal ultrastructural changes were noted with all three types of treatment. The effectiveness of the preparations in resolving the anaemia was similar in the three groups. It was concluded that TM/FMOA exerts a protective effect against the toxicity normally observed of the iron in other formulations in normal and anaemic rats, which was attributed to the fact that administration of iron bound to a protein core allows for gradual release of iron. PMID- 12723895 TI - Potential adverse interaction between aspirin and lisinopril in hypertensive rats. AB - The potential clinical effect of aspirin (ASA) in patients treated with angiotensin converting enzyme (ACE) inhibitors is debatable. Several studies have suggested that ASA attenuates the beneficial effects of ACE inhibitors in hypertension, congestive heart failure (CHF) or coronary artery disease (CAD) and have questioned the safety of using ASA concomitantly with these agents. The present study aims to investigate the possible interaction between ASA and ACE inhibitor in hypertensive rats. Hypertension was induced in adult male Wistar rats using Methylprednisolone (MP) 20 mg/kg per week s.c. for 2 weeks. The systolic blood pressure (SBP) was measured by noninvasive BP technique. The effect of Lisinopril (LS) 15 mg/kg per day and that of combination of LS and ASA; 100 and 25 mg/kg per day p.o. was studied on hypertension induced by glucocorticoid. Concurrent ASA treatment with LS did not hinder the hypotensive effect of LS at either dose. However ASA 100 mg/kg per day caused mortality in animals and produced massive cardiac necrosis and renal damage as evident from histopathology. Treatment with ASA 25 mg/kg per day caused lower mortality with variable effects on cardiac and renal tissues. These results indicate that ASA attenuates the beneficial effects of ACE inhibitor on survival in hypertensive rats and this effect was more pronounced at higher dose of ASA. PMID- 12723896 TI - Effect of leptin administration on plasma and tissue lipids in alcohol induced liver injury. AB - Previous studies suggest a possible link between leptin and hepatic inflammation; however the role of leptin in liver diseases remains unclear. The purpose of the present study was to evaluate the effect of leptin on plasma and tissue lipids in experimental hepatotoxicity. Administering ethanol (6.32 g/kg body weight) to 4 week-old healthy mice for 45 days resulted in significantly elevated levels of plasma and tissue phospholipids, triglycerides and free fatty acids as compared with those of the control animals. Subsequent to the experimental induction of hepatotoxicity (i.e., the initial period of 30 days) exogenous leptin was simultaneously administered (230 microg/kg body weight) every alternate day for 15 days along with the daily dose of alcohol. Leptin administration to control and alcohol-treated mice reduced the weight gain and significantly lowered the levels of plasma and tissue lipids as compared with the untreated control and alcohol supplemented mice. It is postulated that the increase in systemic leptin levels lower the plasma and tissue lipids of alcohol-treated mice, which operates independently of changes in food intake, body weight and the size of the fat stores. PMID- 12723897 TI - The effects of aflatoxin B1 on the development of kwashiorkor in mice. AB - Seventy Swiss albino mice (6-week-old male) were selected for the investigation into aflatoxin B1's role in the cause of kwashiorkor. The mice were divided randomly into four groups. They were grouped within each group by being fed either low or normal protein level diets supplemented with very small amounts of aflatoxin B1 (0.5 microg/day). The control groups were fed aflatoxin B1-free diets containing either normal or low protein levels. All groups were monitored for 7 weeks. The increase in body weight was found to be low in groups I and II, given diets contaminated with aflatoxin B1. Although groups II and IV, which were given low dietary protein, showed remarkable decreases in serum total protein and albumin levels (group II: total protein 4.1 +/- 0.1 g/dL, albumin 2.6 +/- 0.8 g/dL and group IV: total protein 4.6 +/- 1.3 g/dL, albumin 2.8 +/- 0.82 g/dL) when compared with the groups fed a normal dietary protein level (group I: total protein 5.9 +/- 1.3g/dL, albumin 3.4 +/- 0.7g/dL and group III: total protein 5.4 +/- 1.6g/dL, albumin 3.5 +/- 1.2g/dL; P < 0.05). The statistical difference between these two groups was found not to be significant (P > 0.05). However, decreases in total protein and albumin levels were a little more prominent in group II. In addition, histopatological changes of the liver was remarkable in the group fed a low protein diet and aflatoxin B1 when compared with the group fed only a low protein diet and no aflatoxin B1. More significantly, however, was the increase in liver weight in both groups fed a low protein diet (groups II and IV). Our conclusion is that aflatoxin B1 could not have contributed to the development of kwashiorkor. PMID- 12723898 TI - Hair analysis used to assess chronic exposure to the organophosphate diazinon: a model study with rabbits. AB - The main purpose of the present study was to determine whether hair analysis would be a suitable method to assess chronic exposure of rabbits to the pesticide diazinon. A controlled study was designed, in which white rabbits of the New Zealand variety were systemically exposed to two dosage levels (15 mg/kg per day and 8 mg/kg per day) of the pesticide, through their drinking water, for a period of 4 months. Hair samples from the back of the rabbits were removed before commencing the experiment and at the end of the dosing period. Parallel experiments with spiked hair were carried out in order to design a simple and efficient method of extraction of diazinon from hair. The hair was pulverized in a ball mill homogenizer, incubated in methanol at 37 degrees C overnight, liquid liquid extracted with ethyl acetate and measured by chromatography techniques (GC NPD and GC-MS) for confirmation. The concentration of the diazinon in the hair of the exposed animals ranged from 0.11 to 0.26 ng/mg hair. It was concluded that there is a relationship between the administered dose and the detected pesticide concentration in hair. Finally, it seems that hair analysis may be used to investigate chronic exposure to the pesticide. PMID- 12723899 TI - Acute renal failure from organophospate poisoning: a case of success with haemofiltration. AB - Severe organophosphate poisoning (OPP) has a high mortality rate. Respiratory and neurological complications are common in OPP. Multiple organ distress syndrome (MODS) and renal impairment are relatively rare but correlated with death. In previous publications, in patients who did not survive OPP, their deaths were due to MODS or acute renal failure. A case of intentional ingestion of an organophosphate with renal and multiple organ complications is described. In addition to the standard atropine/oxime regimen, continuous venous-venous haemofiltration (CVVH) therapy was started; the patient survived this intoxication. The pathogenesis of renal injury by OPP is unclear and more insight is required. In our experience, CVVH can be a valid therapy, considering in particular the toxicokinetics of the organophosphate. PMID- 12723900 TI - Early substance use by juvenile offenders. AB - Although the interconnection between delinquency and substance use in adolescence is well documented, considerably less is known about substance-use initiation in childhood for juvenile delinquent populations. This descriptive study examined early substance initiation in childhood as reported by adolescents who were incarcerated for juvenile offenses (93 males, 96 females; 58% African American, 42% European American). Youth were individually interviewed using an adapted version of substance-related questions from the National Household Survey. Juvenile justice system records were reviewed to characterize offense histories. A majority of males and females reported using at least one substance (other than cigarettes) such as alcohol, marijuana, or inhalants by age 13. Alcohol use reportedly occurred by age 10 for 17% of the youth. For a substantial portion, early initiation turned into frequent early use. For example, 32% of the males and 39% of the females reported drinking alcoholic beverages at a frequency of several times per month or greater by age 13. Limited evidence related early substance initiation with subsequent substance abuse. Offense status was related to early substance initiation for females but not males. The study provided clear evidence that very early substance use is a significant problem among youth who end up in the juvenile justice system and that we need to find out more about the environmental and social variables affecting very early substance initiation. PMID- 12723901 TI - Gender differences in reactive and proactive aggression. AB - The purpose of our investigation was to study gender differences in proactive and reactive aggression in a sample of 323 clinically referred children and adolescents (68 females and 255 males). Proactive aggression and reactive aggression were assessed using the Proactive/Reactive Aggression Scale. Demographic, historical, family, diagnostic, and treatment variables were entered into stepwise regression analyses to determine correlates of proactive and reactive aggression in males and females. Results reveal high rates of aggression in both males and females in the sample. Self reported drug use, expressed hostility, and experiences of maladaptive parenting were correlated with proactive aggression for both genders. Hyperactive/impulsive behaviors were correlated with male reactive aggression. An early age of traumatic stress and a low verbal IQ were correlated with female proactive aggression. Gender differences in correlates of proactive and reactive aggression may provide possible targets for research, prevention, and treatment efforts focused on reducing maladaptive aggression in clinically referred youth. PMID- 12723902 TI - The stress response in anorexia nervosa. AB - Patients with eating disorders have been found to have problems with Interoceptive Awareness. This study seeks to examine this issue in an experimental paradigm. In the present study, we investigated the hypothesis that, in addition to lowering a body's autonomic stress response, a state of starvation also lowers the psychological stress response. Results indicated that those with anorexia nervosa showed a muted physiology, but they did not show a complete denial of negative emotion. No relation was seen, however, between their affective and physiological responses to a stress task, which contrasted results found for the controls. PMID- 12723903 TI - Defenses in school age children: children's versus parents' report. AB - We tested a questionnaire assessing defenses in school age children and compared the results with their mothers' reports of the same. Thirty-four children (56% male, mean age 8.5) completed the Response Evaluation Measure for Youth (REM-Y) twice over three weeks. Simultaneously, mothers completed the parent version of this measure (REM-P) about their children. In these questionnaires, factor 1 defenses (less adaptive; broken down into intrapersonal and interpersonal defenses) are assessed separately from factor 2, or more adaptive, defenses. Only lower level, interpersonal defenses, such as acting out, were recognized by mothers in their children. The children's intrapersonal defenses, along with factor 2 reactions, were reported less by mothers than by children. PMID- 12723904 TI - Contingency-competence-control-related beliefs and symptoms of anxiety and depression in a young adolescent sample. AB - The present study examined the connection between contingency-competence-control related beliefs, on the one hand, and anxiety and depression, on the other hand, in a large sample of young adolescents aged 10 to 14 years (N = 214). Participants completed measures of perceived contingency, competence, and control, as well as a questionnaire assessing symptoms of common anxiety disorders and major depressive disorder. Results showed that contingency competence-control-related beliefs were intercorrelated and that these beliefs, in turn, were significantly associated with symptoms of anxiety and depression. Structural equation modeling provided support for a model in which perceived contingency and perceived competence predicted perceived control and in which perceived competence (anxiety and depression) and perceived control (depression only), in turn, predicted symptoms of psychopathology. A prospective test of this model indicated that none of the contingency-competence-control-related beliefs was able to predict symptoms of anxiety and depression at 4-weeks follow-up. However, data also demonstrated that perceived competence significantly contributed to the subjective experience of anxiety and depression on both occasions. PMID- 12723905 TI - Reversed-phase liquid chromatographic determination of riboflavin in feeds. AB - A method for determination of riboflavin in animal feeds using liquid chromatography (LC) was developed for feed samples fortified with riboflavin at 1 mg/lb or greater (up to 10,000 mg/lb). Feed samples were extracted in 0.1 N HCl with heating on a steam bath for 30 min, followed immediately by mechanical shaking for 30 min. Sample extracts were diluted to target volume with 2% acetic acid and filtered; riboflavin was determined by LC on a reversed-phase C18 column with 2% acetic acid-acetonitrile (85 + 15) mobile phase for separation and fluorescence detection with excitation at 460 nm and emission at 530 nm. The extraction was compared with that of the AOAC Official Method for riboflavin in food and feed premixes. The 2 method extractions were not significantly different from each other at the 95% confidence level. The developed method also had good linearity over 4 orders of magnitude, recovery of 95-99% from spiked feed samples, a limit of detection of riboflavin at 0.00034 microg/mL in solution, a limit of quantitation of 0.023 mg/lb in feed, and good ruggedness. PMID- 12723907 TI - Spectrofluorimetric and spectrophotometric determination of gliclazide in pharmaceuticals by derivatization with 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole. AB - Accurate, sensitive, and simple spectrophotometric and spectrofluorimetric methods were developed for the determination of gliclazide in pharmaceutical formulations and biological fluids. Both methods are based on a coupling reaction between gliclazide and 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole in borate buffer, pH 7.8, in which a yellow reaction product that can be measured spectrophotometrically at 400 nm was developed. The same product exhibited a yellow fluorescence at 470 nm upon excitation at 400 nm. The absorbance concentration plot was rectilinear over the range of 2-20 microg/mL with minimum detectability [signal-to-noise (S/N) ratio = 2] of 0.2 microg/mL (6.18 x 10(-7) M); the fluorescence-concentration plot was rectilinear over the range of 0.2-2.5 microg/mL with minimum detectability (S/N = 2) of 0.02 microg/mL (6.18 x 10(-8) M). The different experimental parameters affecting the development and stability of the color were carefully studied and optimized. Both methods were successfully applied to the analysis of commercial tablets. The results were in good agreement with those obtained with the official and reference spectrophotometric methods. A proposal of the reaction pathway was presented. PMID- 12723906 TI - Determination of phenolic compounds in dietary supplements and tea blends containing Echinacea by liquid chromatography with coulometric electrochemical detection. AB - Analytical methodologies with ultrasonic extraction and liquid chromatography (LC) were developed for the determination of phenolic compounds in dietary supplements containing Echinacea. The phenolic compounds determined by these methods included caftaric acid, chlorogenic acid, cynarin, echinacoside, and cichoric acid. Samples from tablets, capsules, and bags of tea blends were extracted by sonication for < or = 30 min with methanol-water (60 + 40). The extracts were centrifuged and filtered, and the filtrates were diluted and analyzed by LC using a reversed-phase column and coulometric electrochemical (EC) detection. The mobile phase was acetonitrile-ammonium formate buffer, pH 3.5 (15.3 + 84.7) containing tetrabutyl ammonium hydrogen sulfate as an ion-pairing reagent. Extraction conditions (e.g., composition of the extraction solvent and sonication time) were optimized for different types of samples. Intra- and interday analytical variations were determined, and intraday analyses were performed by 2 independent analysts using 2 different LC systems. Results were generally comparable. The LC method with EC detection showed better sensitivity and selectivity when compared with LC with ultraviolet detection, although results were similar for the 2 methods for major compounds, i.e., caftaric acid, echinacoside, and cichoric acid. The identities of these major compounds found in samples were confirmed by LC/electrospray ionization mass spectrometry. PMID- 12723908 TI - Simultaneous determination of eleven dyes and their aluminum lakes in drugs. AB - A 3-step extraction method was developed for the simultaneous determination of 11 dyes and their aluminum lakes in drugs. The dyes were first extracted with warm water (approximately 60 degrees C) and were cleaned up by solid-phase extraction with a tC18 cartridge. Aluminum lake dyes that remained in the precipitate were extracted with 0.02M NaOH. Aluminum in the dye lakes was reextracted into the organic layer with acetylacetone-butyl acetate (1 + 9, v/v), as an acetylacetone chelate, and was quantified by atomic absorption spectrometry. The dye portions of the aluminum lakes remained in the aqueous layer and were cleaned up in the same way as the dyes. The dyes and the dye portions of the aluminum lakes were quantified by ion-pair liquid chromatography with a photodiode array detector within 20 min. The recoveries of dyes from drug fortified at 10 microg of each dye per pill were 87.0-102.2%, and the recoveries of dyes from drugs fortified at 50 microg of each dye lake per pill were 82.9-101.6%, except for recoveries of indigo carmine. In 40 ethical and over-the-counter drugs, dyes that were not indicated in the package insert information for drugs were detected in 5 samples. The highest amount of dye found in a drug was 1169.5 microg erythrosine, which was detected in a capsule of antibiotic. Aluminum lake dyes were detected in 8 samples of various dosage forms. PMID- 12723909 TI - Concentration of sodium ion as the determining factor for the association of dansyl amino acids with the teicoplanin molecule in reversed-phase liquid chromatography. AB - The mechanism of the binding of D,L dansyl amino acids to teicoplanin was investigated. Na+ was used as an indicator of the interactions between the solutes and teicoplanin. The number (n) of sodium ions, Na+, excluded from the solute-teicoplanin interface when analyte transfer occurred was determined. A thermodynamic study and enthalpy-entropy compensation were performed to further explore the interaction mechanism. From these results, it was shown that teicoplanin was balanced between 2 conformational states characterized by distinct enantioselective properties. This approach indicates that liquid chromatography (LC) is a useful tool to extract physicochemical and molecular information from retention data. Thus, LC can be used as a complementary technique with the conventional techniques of molecular interaction analysis. PMID- 12723910 TI - Determination of fleroxacin in pure and tablet forms by liquid chromatography and derivative UV-spectrophotometry. AB - Derivative UV-spectrophotometric and liquid chromatographic (LC) methods for fleroxacin determination were validated. In the spectrophotometric assay, first-, second-, third-, and fourth-order measurements were applied with the use of peak zero and peak-peak techniques. The linear correlation between amplitude of the peak and concentration of the examined drug ranged from 2.0 to 12.0 micro/mL. An isocratic LC analysis was performed on a Purospher ODS column with an acidic mobile phase containing tetrabutylammonium hydroxide. Measurements were made at a wavelength of 285 nm with 4-aminobenzoic acid (PABA) as internal standard. The calibration curve was linear (r = 0.9999) in the studied range of concentration (1.0-10.0 microg/mL). The accuracy (mean recovery, about 100%), precision (relative standard deviation < 1%), selectivity, and sensitivity of the elaborated methods were satisfactory. PMID- 12723911 TI - Simultaneous determination of different antibiotic residues in bovine and in porcine kidneys by solid-phase fluorescence immunoassay. AB - Parallux, a solid-phase fluorescence immunoassay (SPFIA) developed for antibiotic residue detection in milk, was used for analysis of bovine and porcine kidney tissue. Four tetracyclines, 2 broad-spectrum cephalosporins, 3 beta-lactam antibiotics, and cephapirin were detected in one run after minimal sample preparation. This commercially available test system is designed as cartridges, each with a combination of 1-4 tests. One cartridge can be used to detect 4 analytes in the same sample, or 1 or 2 analytes in different samples. The cartridge with the combination tetracyclines-ceftiofur-penicillin-cephapirin was selected because tetracyclines, beta-lactam antibiotics as well as cephalosporins, are registered for oral or parenteral use in bovines and pigs in Europe. The test is qualitative and is recommended only for screening. Tetracycline, oxytetracycline, chlortetracycline, and doxycycline were easily detected at 300 ppb with the tetracyclines channel; ceftiofur at 1000 ppb and cefquinome at 200 ppb with the ceftiofur channel; penicillin G, ampicillin, and amoxicillin at 50 ppb with the penicillin channel; and cephapirin at 100 ppb with the cephapirin channel. These levels are equal to or lower than the corresponding maximal residue limits in kidney tissue. Cephalexin was not detected. The SPFIA test can be used as an alternative to classical inhibition tests and for post screening inhibitor- positive kidneys, because it detects 3 specific groups of antibiotics, which enables selection of specific confirmatory methods for identification and quantification. PMID- 12723912 TI - Simultaneous determination of dapsone and pyrimethamine by derivative spectrophotometry in pharmaceutical formulations. AB - A simple and fast method was developed for the simultaneous determination of dapsone and pyrimethamine by first-order digital derivative spectrophotometry. Acetonitrile was used as a solvent to extract the drugs from the pharmaceutical formulations, and the samples were subsequently evaluated directly by digital derivative spectrophotometry. The simultaneous determination of both drugs was performed by the zero-crossing method at 249.4 and 231.4 nm for dapsone and pyrimethamine, respectively. The best signal-to-noise ratio was obtained when the first derivative of the spectrum was used. The linear range of determination for the drugs was from 6.6 x 10(-7) to 2.0 x 10(-4) and from 2.5 x 10(-6) to 2.0 x 10(-4) mol/L for dapsone and pyrimethamine, respectively. The excipients of commercial pharmaceutical formulations did not interfere in the analysis. Chemical and spectral variables were optimized for determination of both analytes. A good level of repeatability, 0.6 and 1.7% for dapsone and pyrimethamine, respectively, was observed. The proposed method was applied for the simultaneous determination of both drugs in pharmaceutical formulations. PMID- 12723913 TI - Determination of semivolatile organic compounds in environmental samples by gas chromatography/mass spectrometry after extraction by cyclic steam distillation. AB - A method was developed for the multiple determination of semivolatile organic compounds found in groundwater, river water, seawater, sediment, and soil. Forty standard compounds were determined: n-alkenes, cycloalkanes, aromatic hydrocarbons, and polycyclic aromatic hydrocarbons. The compounds were isolated from water and soil samples by using an essential oil distillator (cyclic steam distillator) with hexane as a solvent. The extract was cleaned by using a silica gel cartridge with an acetone-hexane solution. The compounds were determined by using a gas chromatograph/mass spectrometer with 12 stable isotope-labeled compounds (surrogate compounds). The efficiencies of recoveries from water samples were 80.0-106% for groundwater, 80.1-106% for river water, and 81.2-103% for seawater. The relative standard deviation (RSD) values were 2.05-16.0% for groundwater, 3.22-16.6% for river water, and 4.45-16.0% for seawater. The efficiencies of recoveries from sediment and soil were 71.5-96.4% and 70.1-99.8%, respectively. RSD values ranged from 2.27 to 16.0% for sediment and from 2.12 to 15.1% for soil. Adjustment of recovery efficiencies of standard compounds by using surrogate compounds gave more accurate values. The present study proved that an essential oil distillator provides satisfactory results for multiple determinations of the semivolatile compounds in environmental waters, sediment, and soil. PMID- 12723914 TI - Enumeration of total aerobic microorganisms in foods by SimPlate Total Plate Count-Color Indicator methods and conventional culture methods: collaborative study. AB - The relative efficacy of the SimPlate Total Plate Count-Color Indicator (TPC-CI) method (SimPlate 35 degrees C) was compared with the AOAC Official Method 966.23 (AOAC 35 degrees C) for enumeration of total aerobic microorganisms in foods. The SimPlate TPC-CI method, incubated at 30 degrees C (SimPlate 30 degrees C), was also compared with the International Organization for Standardization (ISO) 4833 method (ISO 30 degrees C). Six food types were analyzed: ground black pepper, flour, nut meats, frozen hamburger patties, frozen fruits, and fresh vegetables. All foods tested were naturally contaminated. Nineteen laboratories throughout North America and Europe participated in the study. Three method comparisons were conducted. In general, there was <0.3 mean log count difference in recovery among the SimPlate methods and their corresponding reference methods. Mean log counts between the 2 reference methods were also very similar. Repeatability (Sr) and reproducibility (SR) standard deviations were similar among the 3 method comparisons. The SimPlate method (35 degrees C) and the AOAC method were comparable for enumerating total aerobic microorganisms in foods. Similarly, the SimPlate method (30 degrees C) was comparable to the ISO method when samples were prepared and incubated according to the ISO method. PMID- 12723915 TI - Detection of Salmonella in fresh cheese, poultry products, and dried egg products by the ISO 6579 Salmonella culture procedure and the AOAC official method: collaborative study. AB - Three food types were analyzed for the presence of Salmonella by the AOAC culture method and by the International Organization for Standardization (ISO 6579:2002) culture method. Paired test portions of each food type were simultaneously analyzed by both methods. A total of 21 laboratories representing federal government agencies and private industry, in the United States and Europe, participated in this interlaboratory study. Foods were artificially contaminated with Salmonella and competing microflora if naturally contaminated sources were not available. No statistical differences (p < 0.05) were observed between the AOAC and ISO culture methods for fresh cheese and dried egg products. A statistically significant difference was observed for one of the 2 lots of poultry from the first trial. The poultry meat used in this run was radiation sterilized, artificially contaminated with Salmonella and competitive flora, and then lyophilized. A second trial was conducted with 2 separate lots of raw ground chicken that were naturally contaminated. The results from the second trial showed no statistical difference between the 2 culture methods. A third trial involving 4 laboratories was conducted on 2 separate lots of naturally contaminated raw poultry. Again, no statistically significant differences occurred. It is recommended that ISO 6579:2002 culture method for Salmonella be adopted Official First Action for the analysis of fresh cheese, fresh chilled and frozen poultry, and dried egg products. PMID- 12723916 TI - Enumeration of total yeasts and molds in foods by the SimPlate Yeast and Mold Color Indicator method and conventional culture methods: collaborative study. AB - The relative effectiveness of the SimPlate Yeast and Mold-Color Indicator method (Y&M-CI) was compared to the U.S. Food and Drug Administration's (FDA) Bacteriological Analytical Manual (BAM) method and the proposed International Organization for Standardization (ISO) method, ISO/CD 21527, for enumerating yeasts and molds in foods. Test portions were prepared and incubated according to the conditions stated in both the BAM and ISO methods. Six food types were analyzed: frozen corn dogs, nut meats, frozen fruits, cake mix, cereal, and fresh cheese. Nut meats, frozen fruits, and fresh cheese were naturally contaminated. All other foods were artificially contaminated with either a yeast or mold. Seventeen laboratories throughout North America and Europe participated in the study. Three method comparisons were conducted. In general, there was <0.3 mean log count difference in recovery between the SimPlate method and the 2 corresponding reference methods. Moreover, mean log counts between the 2 reference methods were also very similar. The repeatability (Sr) and reproducibility (SR) standard deviations were comparable between the 3 method comparisons. These results indicate that the BAM method and the SimPlate method are equivalent for enumerating yeast and mold populations in foods. Similarly, the SimPlate method is comparable to the proposed ISO method when test portions are prepared and incubated as defined in the proposed ISO method. PMID- 12723917 TI - Detection of botulinal neurotoxins A, B, E, and F by amplified enzyme-linked immunosorbent assay: collaborative study. AB - An amplified enzyme-linked immunosorbent assay (amp-ELISA) was compared with the AOAC Official Method 977.26 for detection of Clostridium botulinum and its toxins in foods. Eleven laboratories participated and the results of 10 laboratories were used in the study. Two anaerobic culture media, tryptone peptone glucose yeast extract (TPGY) and cooked meat medium (CMM) were used to generate toxic samples with types A, B, E, and F botulinal strains. Nonbotulinal clostridia were also tested. The toxicity of each botulinal culture was determined by the AOAC method, and the cultures were then diluted, if necessary, to high (about 10,000 minimal lethal dose [MLD]/mL) and low (about 100 MLD/mL) test samples. The overall sensitivity of detection in TPGY and CMM cultures with the amp-ELISA was 94.7% at about 100 MLD/mL and 99.6% for samples with > or = 10,000 MLD/mL toxicity. The amp-ELISA detection sensitivity for low toxin samples was 92.3% in TPGY and 99.4% in CMM. The false-positive rate ranged from 1.5% for type A to 28.6% for type F in TPGY, and from 2.4% for type A to 11.4% for type F in CMM. Most of the cross-reactivity was due to detection of other botulinal types, especially in high toxin samples. The amp-ELISA could be used to screen suspect cultures for botulinal toxins. Positive amp-ELISA samples would be confirmed by the AOAC reference method. PMID- 12723918 TI - Modification of enrichment protocols for TECRA Listeria Visual Immunoassay method 995.22: collaborative study. AB - A collaborative study was conducted to validate new enrichment methods for the TECRA Listeria Visual Immunoassay (TLVIA). These new methods incorporate a newly formulated medium, TECRA Listeria Enrichrment Broth, which does not contain the highly toxic antifungal agent, cycloheximide. The new procedures will provide an alternative to the enrichment procedures described in AOAC Method 995.22. Three food types (raw ground beef, lettuce, and ice cream) were analyzed in the United States, and 2 food types (cooked turkey and cooked fish fillets) were analyzed in Australasia. Thirty collaborators participated in the study, 16 in Australasia and 14 in the United States. With the exception of one batch of ground beef, comparison of the proportion of positive test portions (p > or = 0.05) showed no significant difference between the TLVIA and the reference method for the 5 foods at 3 inoculation levels. For the one batch of naturally contaminated raw ground beef, the TLVIA gave significantly more confirmed positive results than the reference method. PMID- 12723919 TI - Sensitivity and specificity of the Sanita-kun Aerobic Count: internal validation and independent laboratory study. AB - The Sanita-kun Aerobic Count consists of a transparent cover film, an adhesive sheet, a layer of nonwoven fabric, and a water-soluble compound film, including a culture medium formula for detection of aerobic microorganisms. The Sanita-kun sheet was validated for 14 food categories in an internal study and an independent study was conducted on ground beef and hot dogs. Both studies showed no significant difference in performance between 5 or 8 replicates of the Sanita kun sheets and AOAC Method 966.23, excluding some lots of foods. The correlation coefficient to plate count agar in the internal accuracy study was 0.99. The average relative standard deviation for repeatability of total foods was 0.26 and 0.19, respectively, excluding < 10 average counts. The ruggedness study, which examined the influence of incubation temperature and period, recommended incubation of the Sanita-kun sheet at 32.5 +/- 2.5 degrees C for 46 +/- 2 h. Comparison of 3 lots of Sanita-kun sheets showed no decrease of performance in the older lot. The shelf-life of the sheet is at least 14 months. The Sanita-kun Aerobic Counts has been granted AOAC Performance Tested Method status. PMID- 12723920 TI - Determination of total vitamin C in fruit juices and related products by liquid chromatography: interlaboratory study. AB - A interlaboratory study was conducted to evaluate a liquid chromatographic (LC) procedure for the determination of total vitamin C in foods at levels of 5-60 mg/100 g. Emphasis was placed on fruit juices, although selected foods were also included in the study. Following dissolution of sample in water, endogenous dehydroascorbic acid was converted to ascorbic acid by precolumn reduction with dithiothreitol at neutral pH. Total ascorbate was determined by C18 reversed phase LC with a phosphate eluent at pH 2.5, incorporating dithiothreitol to maintain vitamin C in the reduced form, and UV detection at 254 nm. Seven types of fruit juices and foods were tested by 19 collaborators in 7 countries. Three duplicate juices and foods met the criteria for Youden pairs and yielded repeatability relative standard deviation of 5.80-14.66%. Reproducibility relative standard deviation ranged from 6.36 to 35.54% (n = 10) with HORRAT values of 0.82-4.04. The LC method is suitable for routine use in fruit products and foods containing > 5 mg/100 g vitamin C and is recommended for further validation by AOAC INTERNATIONAL and International Fruit Juice Union. PMID- 12723921 TI - Determination of retinyl palmitate (vitamin A) in fortified fluid milk by liquid chromatography: collaborative study. AB - A liquid chromatographic (LC) method was developed for fast and simple measurement of retinyl palmitate (vitamin A) in fortified milk. Retinyl acetate internal standard was added to a test portion of milk followed by extraction into hexane. The hexane extract was analyzed by LC using a normal-phase silica gel column equilibrated with mobile phase (conditioned hexane-isopropanol, 99.85 + 0.15, v/v) about 1 h before injections. The retinyl palmitate concentration was calculated by using a relative response factor determined with calibration standards. In the collaborative study, 11 laboratories analyzed 13 pairs of fluid milk materials in blind duplicate. Twelve of the materials were composed of skim milk (< 0.5% fat), 1% fat milk, 2% fat milk, and 1% fat chocolate milk. Each material was fortified at 3 concentrations of retinyl palmitate of approximately 581 microg/L (1000 IU/qt), 1163 microg/L (2000 IU/qt), and 2236 microg/L (4000 IU/qt). The 13th material, unfortified skim milk, served as a matrix blank. Repeatability standard deviations (RSDr) without outliers ranged from 1.5 to 5.7% and reproducibility standard deviations (RSDR) without outliers ranged from 5.0 to 22.7%. cis-Isomers co-eluted with the predominant trans-retinyl palmitate isomer and were included in the results reported by all the collaborative laboratories. Endogenous long-chain esters from milk fat were also measured with the retinyl palmitate additive. The Study Director recommends that this method for determination of retinyl palmitate in fluid milk by LC be adopted First Action. PMID- 12723922 TI - Determination of immunoglobulin G in bovine colostrum and milk by direct biosensor SPR-immunoassay. AB - An automated biosensor surface-plasmon resonance-based assay was developed for the determination of immunoglobulin G (IgG) in bovine milk and colostrum with either goat or rabbit antibovine IgG or protein G used as detecting molecule. The method is configured as a direct and nonlabeled immunoassay, with quantitation against an authentic IgG calibrant. Whole colostrum or milk is prepared for analysis by dilution into buffer. Analysis conditions, including ligand immobilization, flowrate, contact time, and regeneration, were optimized, and nonspecific binding was evaluated. Performance parameters included working range of 15-10 000 ng/mL, method detection limit of 0.08 mg/mL, overall instrument response reproducibility relative standard deviation (RSD(R)) of 0.47%, mean between-run RSD(R) of 10.5% for colostrum, and surface stability over 200 analyses. The proposed method was compared with independent alternative methods. The technique was applied to the measurement of IgG content during early lactation transition from colostrum to milk, as well as in consumer milk, colostrum, and hyperimmune milk powders. PMID- 12723923 TI - Analytical pervaporation: a key technique in the enological laboratory. AB - This paper reviews the use of analytical pervaporation (defined as the integration of 2 different analytical separation principles, evaporation and gas diffusion, in a single micromodule) coupled to flow-injection manifolds for the determination of analytes of interest in enology; the review discusses the advantages that these techniques can provide in wine analytical laboratories. Special attention is given to methods that enable the determination of either of 2 volatile analytes, or of one volatile analyte and one nonvolatile analyte by taking advantage of the versatility of the designed approaches. In a comparison of these methods with the official and/or standard methods, the results showed good agreement. In addition, the new methods offer improvements in linear determination range, quantitation limit, precision, rapidity, and potential for full automation. Thus, this review demonstrates that although the old technologies used in wine analytical laboratories may be supported by official and standard methods, they should be replaced by properly validated, new, and automated technologies. PMID- 12723924 TI - Determination of cholecalciferol (vitamin D3) in selected foods by liquid chromatography: NMKL collaborative study. AB - Results are presented from an NMKL (Nordic Committee on Food Analysis) collaborative study of a method for the determination of cholecalciferol (vitamin D3) in foods. The method is based on the addition of an internal standard (vitamin D2), followed by saponification and extraction with n-heptane. The fraction that contains vitamin D2/D3 is separated by preparative normal-phase liquid chromatography (LC), and the analytes are determined by reversed-phase LC with UV detection at 265 nm. The method was tested by 8 participating laboratories. In this study 6 different matrixes were analyzed for cholecalciferol content: milk, liquid infant formula (gruel), cooking oil, margarine, infant formula, and fish oil. The contents varied from 0.4 to 12 microg/100 g. Three matrixes (milk, gruel, and margarine) were fortified with vitamin D3. In the other matrixes, vitamin D3 was added at 3 different levels at the Swedish National Food Administration. The milk was analyzed as a blind duplicate, whereas the other matrixes were analyzed as split-level pairs. The recoveries from the samples with vitamin D3 added varied from 93 to 102%. The repeatability relative standard deviation (RSDr) values for accepted results varied between 2.2% (fish oil) and 7.4% (cooking oil), whereas the reproducibility relative standard deviation (RSD(R)) values varied between 6.8% (margarine) and 24% (cooking oil). PMID- 12723925 TI - Alternative AOAC sporicidal test carrier for evaluating peracetic acid-based sterilants (modification of AOAC official method 966.04). AB - Dacron suture loops were demonstrated to be inert, consistent carriers in the presence of peracetic acid-based sterilants, whereas black silk sutures had a variable preparation process and interacted with peracetic acid. In addition, Dacron suture loops provided comparable spore loading to black silk suture loops and an HCI resistance of > or = 2 min. These results indicate that black silk suture loops are not appropriate carriers for assessing peracetic acid-based sterilants, and Dacron loops are an acceptable alternative. This finding is consistent with the Office of Science and Technology Laboratory (Center for Devices and Radiological Health) study which determined that "polyester suture material is a viable alternative to silk for the AOAC sporicidal test for liquid disinfectants." PMID- 12723926 TI - Fast and easy multiresidue method employing acetonitrile extraction/partitioning and "dispersive solid-phase extraction" for the determination of pesticide residues in produce. AB - A simple, fast, and inexpensive method for the determination of pesticide residues in fruits and vegetables is introduced. The procedure involves initial single-phase extraction of 10 g sample with 10 mL acetonitrile, followed by liquid-liquid partitioning formed by addition of 4 g anhydrous MgSO4 plus 1 g NaCl. Removal of residual water and cleanup are performed simultaneously by using a rapid procedure called dispersive solid-phase extraction (dispersive-SPE), in which 150 mg anhydrous MgSO4 and 25 mg primary secondary amine (PSA) sorbent are simply mixed with 1 mL acetonitrile extract. The dispersive-SPE with PSA effectively removes many polar matrix components, such as organic acids, certain polar pigments, and sugars, to some extent from the food extracts. Gas chromatography/mass spectrometry (GC/MS) is then used for quantitative and confirmatory analysis of GC-amenable pesticides. Recoveries between 85 and 101% (mostly > 95%) and repeatabilities typically < 5% have been achieved for a wide range of fortified pesticides, including very polar and basic compounds such as methamidophos, acephate, omethoate, imazalil, and thiabendazole. Using this method, a single chemist can prepare a batch of 6 previously chopped samples in < 30 min with approximately 1 dollar (U.S.) of materials per sample. PMID- 12723927 TI - Comparative tests to improve the gas chromatographic analysis of chlorobornanes in fish samples. AB - A comparison was made between electron capture negative ionization quadrupole mass spectrometry (ECNI-MS) and electron capture detection (ECD) with regard to repeatability and reproducibility for the gas chromatographic (GC) analysis of toxaphene congeners [chlorobornanes (CHBs)]. The tests, including standard solutions and several cleaned fish extracts, showed larger relative standard deviations (RSDs) for the repeatability of ECNI-MS but no differences in the reproducibility of the 2 techniques. The sensitivity of the GC-ECNI-MS was considerably better than that of GC/ECD. Four stepwise-designed comparative tests were also conducted on GC analysis, cleanup, and the complete method. The results showed that, according to the current state-of-the-art, coefficients of variation for the between-laboratory performance were not < 20% and were usually between 20 and 30%. In spite of separation problems, e.g., for CHB 26, which cannot be separated into a single-component peak, a 95% methyl 5% phenyl polysiloxane (CP Sil 8) column was preferred to more polar columns for the analysis of CHBs 26, 40, 41, 44, 50, and 62. CHB 62 was more difficult to determine than CHB 26 and 50. Addition of the CHBs 40, 41, and 44 to the standard set of 3 chlorobornanes (26, 50, and 62) resulted in more separation problems. A 3-step cleanup method was recommended. PMID- 12723929 TI - Antisense DNAs as targeted genetic medicine to treat cancer. AB - Nucleic acid therapies represent a direct genetic approach for cancer treatment. Such an approach takes advantage of mechanisms that activate genes known to confer a growth advantage to neoplastic cells. The ability to block the expression of these genes allows exploration of normal growth regulation. Progress in antisense technology has been rapid, and the traditional antisense inhibition of gene expression is now viewed on a genomic scale. This global view has led to a new vision in antisense technology, the elimination of nonspecific and undesirable side effects, and ultimately, the generation of more effective and less toxic nucleic acid medicines. Several antisense oligonucleotides are in clinical trials, are well tolerated, and are potentially active therapeutically. Antisense oligonucleotides are promising molecular medicines for treating human cancer in the near future. PMID- 12723928 TI - Determination of metals in composite diet samples by inductively coupled plasma mass spectrometry. AB - A study was conducted to evaluate the applicability of inductively coupled plasma mass spectrometry (ICP-MS) techniques for determination of metals in composite diets. Aluminum, cadmium, chromium, copper, lead, manganese, nickel, vanadium, and zinc were determined by this method. Atmospheric pressure microwave digestion was used to solubilize analytes in homogenized composite diet samples, and this procedure was followed by ICP-MS analysis. Recovery of certified elements from standard reference materials ranged from 92 to 119% with relative standard deviations (RSDs) of 0.4-1.9%. Recovery of elements from fortified composite diet samples ranged from 75 to 129% with RSDs of 0-11.3%. Limits of detection ranged from 1 to 1700 ng/g; high values were due to significant amounts of certain elements naturally present in composite diets. Results of this study demonstrate that low-resolution quadrupole-based ICP-MS provides precise and accurate measurements of the elements tested in composite diet samples. PMID- 12723930 TI - Synthesis and immunosuppressive activity of novel succinylacetone analogues. AB - This study describes the synthesis of novel enol esters (3) and triketones (4) as analogues of succinylacetone (SA) (Ed- this abbreviation is introduced here based on your use of it in the body of the paper) and the evaluation on the mouse allogeneic mixed lymphocyte reaction (MLR) and the murine model of antigen induced paw edema formation for immunosuppressive activity. Enol esters (3a-f) were about 2-4 fold more potent than SA in in vitro activity. PMID- 12723931 TI - Synthesis of beta-hydroxy-propenamide derivatives and the inhibition of human dihydroorotate dehydrogenase. AB - Novel beta-hydroxy propenamides as analogues of the active metabolite of leflunomide (A 771726) were synthesized and evaluated for their inhibitory activity on dihydroorotate dehydrogenase (DHODH) in an investigation into their immunosuppressive activity. Compounds 2a, 3a, and 3h were approximately 4-40 times more potent than leflunomide in their activity while they were-less active than A 771726. PMID- 12723932 TI - Synthesis and antifungal activities of some aryl (3-methyl-benzofuran-2-yl) ketoximes. AB - In this study, some aryl (3-methyl-benzofuran-2-yl) ketoximes and their ethers and esters were synthesised. The structure elucidation of the compounds was performed by IR, 1H-NMR, MASS spectroscopy and elemental analyses. Antifungal activities of the compounds were examined and moderate activity was obtained. PMID- 12723933 TI - A new (E)4-hydroxy-dodec-2-enedioic acid from the stem bark of Albizzia julibrissin. AB - A new unsaturated hydroxy acid was isolated from the stem bark extract of Albizzia julibrissin through repeated silica gel and Sephadex LH-20 column chromatography. The chemical structure of the new acid was determined as (E)-4 hydroxy-dodec-2-enedioic acid on the basis of several spectral data including 2D NMR. The stereochemical feature of the double bond was determined to be E on the basis of the coupling pattern of related proton signals in the 1H-NMR and COSY experiments. PMID- 12723934 TI - Study of substance changes in flowers of Pueraria thunbergiana Benth. during storage. AB - Puerariae Flos is a traditional herbal medicine that has long been used as a treatment for colds, diabetes, and hangovers. The herbal medicine contains a wide variety of isoflavones such as kakkalide, tectoridin, and tectorigenin. This study demonstrates that the substances undergo a certain degree of change depending on the storage period by the method of HPLC and 13C-NMR, and that the HPLC analysis can be used to determine the freshness of Puerariae Flos. PMID- 12723935 TI - Comparison of green tea extract and epigallocatechin gallate on blood pressure and contractile responses of vascular smooth muscle of rats. AB - The present study was conducted to investigate the effects of green tea extract (GTE) on arteral blood pressure and contractile responses of isolated aortic strips of the normotensive rats and to establish the mechanism of action. The phenylephrine (10(-8) approximately 10(-5) M)-induced contractile responses were greatly inhibited in the presence of GTE (0.3 approximately 1.2 mg/mL) in a dose dependent fashion. Also, high potassium (3.5 x 10(-2) approximately 5.6 x 10(-2) M)-induced contractile responses were depressed in the presence of 0.6 approximately 1.2 mg/mL of GTE, but not affected in low concentration of GTE (0.3 mg/mL). However, epigallocatechin gallate (EGCG, 4 approximately 12 microg/mL) did not affect the contractile responses evoked by phenylephrine and high K+. GTE (5 approximately 20 mg/kg) given into a femoral vein of the normotensive rat produced a dose-dependent depressor response, which is transient. Interestingly, the infusion of a moderate dose of GTE (10 mg/kg/30 min) made a significant reduction in pressor responses induced by intravenous norepinephrine. However, EGCG (1 mg/kg/30 min) did not affect them. Collectively, these results obtained from the present study demonstrate that intravenous GTE causes a dose-dependent depressor action in the anesthetized rat at least partly through the blockade of adrenergic alpha1-receptors. GTE also causes the relaxation in the isolated aortic strips of the rat via the blockade of adrenergic alpha1-receptors, in addition to the unknown direct mechanism. It seems that there is a big difference in the vascular effect between GTE and EGCG. PMID- 12723936 TI - LB30057, an orally effective direct thrombin inhibitor, prevents arterial and venous thrombosis in rats and dogs. AB - The anti-thrombotic effects of LB30057, a direct thrombin inhibitor, were evaluated with in vivo rat and dog thrombosis models. In rats, 1 mg/kg of LB30057 inhibited half of the clot formations in the inferior vena cava at 5 minutes after intravenous application. When measured at 2 hours after oral application, 100 mg/kg prevented approximately half of the clot formations in the inferior vena cava and 50 mg/kg prolonged the mean occlusion time from 15.6 +/- 1.3 minutes to 47.2 +/- 8.3 minutes in the carotid artery. In dogs, the formation of thrombus in the jugular vein was reduced to half at a dose range of 20-30 mg/kg at 6 hours after oral application. In addition, the LB30057 dosage required to reduce venous clot formation by approximately 80-90% in dogs was only about 10% of that required for the same reduction in rats. This is probably due to the variation in its time-dependent blood concentration profiles in each species; for example, the plasma half-life of LB71350 in dogs was longer than that in rats (153.0 +/- 3.0 vs. 129.7 +/- 12.7 min at 30 mg/kg, i.v., respectively). AUC, T(max), C(max), and BA in dogs were 59, 8.9, 9.17, and 13.3 times higher than those in rats at oral 30 mg/kg, respectively. Taken together, these results suggest that LB30057 administered orally is effective in the prevention of arterial and venous thrombosis in rats and dogs. It therefore represents a good lead compound for investigations to discover a new, orally available, therapeutic agent for treating thrombotic diseases. PMID- 12723938 TI - The role of ascorbic acid on the redox status and the concentration of malondialdehyde in streptozotocin-induced diabetic rats. AB - We investigated the role of ascorbic acid on the redox status in streptozotocin induced diabetic rats. In the plasma of diabetic rats, the ratio of reduced/total ascorbic acid was significantly decreased as compared with normal control. Ascorbic acid supplementation increased the reduced and total ascorbic acid contents as compared with diabetic control. In the rutin-treatment group, reduced and total contents of ascorbic acid were significantly decreased, however, the ratio of reduced/total contents of ascorbic acid had no difference as compared with diabetic rats. In the insulin-treatment group, this ratio is not significantly different as compared with diabetic control. However, in the insulin plus ascorbic acid treatment group, reduced form and the ratio of reduced/total ascorbic acid were significantly increased as compared with diabetic control. In addition, we measured the contents of malondialdehyde (MDA) in the plasma of diabetic rats. The contents of MDA was increased as compared with normal control, however, in insulin-treatment group, the contents of MDA was decreased as compared with diabetic rats. Ascorbic acid had no effects on the increases of MDA in diabetic rats. In conclusion, plasma ascorbic acid level and its reduced/total ratio reflects the status of the oxidative stress in the diabetic rats. Supplement of ascorbic acid did not correct the ratio of the reduced/ total ascorbic acid. However, supplement of insulin and ascorbic acid corrected the ratio of reduced/total ascorbic acid. PMID- 12723937 TI - Anti-inflammatory activity of Elsholtzia splendens. AB - Elsholtzia splendens Nakai has been used in North-East Asia as an ingredient of folk medicines for treating cough, headache and inflammation. The present investigation was carried out to establish its in vivo anti-inflammatory activity using several animal models of inflammation and pain. The 75% ethanol extract of the aerial part of E. splendens significantly inhibited mouse croton oil-induced, as well as arachidonic acid-induced, ear edema by oral administration (44.6% inhibition of croton oil-induced edema at 400 mg/kg). This plant material also showed significant inhibitory activity against the mouse ear edema induced by multiple treatment of phorbol ester for 3 days, which is an animal model of subchronic inflammation. In addition, E. splendens exhibited significant analgesic activity against mouse acetic acid-induced writhing (50% inhibition at 400 mg/kg), while indomethacin (5 mg/kg) demonstrated 95% inhibition. E. splendens (5-100 microg/mL) significantly inhibited PGE2 production by pre induced cyclooxygenase-2 of lipopolysaccharide-treated RAW 264.7 cells, suggesting that cyclooxygenase-2 inhibition might be one of the cellular mechanisms of anti-inflammation. PMID- 12723939 TI - Modulation of TNF-alpha-induced ICAM-1 expression, NO and H2O2 production by alginate, allicin and ascorbic acid in human endothelial cells. AB - Plant nutrients are believed to provide protection against various diseases including inflammation. Since interactions of the cell adhesion molecules are known to play important roles in mediating inflammation, inhibiting adhesion protein upregulation is a possible therapeutic target. In this study, the interacellular adhesion molecule-1 (ICAM-1) was induced in human umbilical endothelial cells (HUVECs) after stimulation with TNF-alpha. In addition, alginate, ascorbic acid and allicin were demonstrated to inhibit the TNF-alpha induced expression of ICAM-1 on the HUVECs in a dose-dependent manner. These compounds also inhibited the production of NO and H2O2 induced by TNF-alpha, which suggests that the inhibition of ICAM-1 expression by the three compounds may be due to the modulated production of the reactive oxygen/nitrogen components. Overall, these results indicate that these dietary components have a therapeutic potential in the treatment of various inflammatory disorders associated with an increase in endothelial leukocyte adhesion molecules. PMID- 12723940 TI - 2002 Medicinal Chemistry Division Award address: monoamine transporters and opioid receptors. Targets for addiction therapy. PMID- 12723941 TI - Synthesis and antibacterial activity of 6-O-arylbutynyl ketolides with improved activity against some key erythromycin-resistant pathogens. AB - A series of novel 6-O-substituted homopropargyl ketolides was synthesized and evaluated against various erythromycin-resistant pathogens. Promising in vitro antibacterial activity was demonstrated for compounds bearing this structural motif. PMID- 12723942 TI - Design and synthesis of statine-based cell-permeable peptidomimetic inhibitors of human beta-secretase. AB - We describe the development of statine-based peptidomimetic inhibitors of human beta-secretase (BACE). The conversion of the peptide inhibitor 1 into cell permeable peptidomimetic inhibitors of BACE was achieved through an iterative strategy of conceptually subdividing 1 into three regions: an N-terminal portion, a central statine-containing core, and a C-terminus. Replacement of the amino acid residues of 1 with moieties with less peptidic character was done with retention of BACE enzyme inhibitory activity. This approach led to the identification of the cell-permeable BACE inhibitor 38 that demonstrated BACE mechanism-selective inhibition of Abeta secretion in human embryonic kidney cells. PMID- 12723943 TI - Benzodiazepines as potent and selective bradykinin B1 antagonists. AB - Antagonism of the bradykinin B(1) receptor was demonstrated to be a potential treatment for chronic pain and inflammation. Novel benzodiazepines were designed that display subnanomolar affinity for the bradykinin B(1) receptor (K(i) = 0.59 nM) and high selectivity against the bradykinin B(2) receptor (K(i) > 10 microM). In vivo efficacy, comparable to morphine, was demonstrated for lead compounds in a rodent hyperalgesia model. PMID- 12723944 TI - Retroinverso analogue of the antiviral octapeptide C8 inhibits feline immunodeficiency virus in serum. AB - We described the antiviral activity of an octapeptide corresponding to a Trp-rich domain of feline immunodeficiency virus (FIV) transmembrane glycoprotein. To overcome the limited enzymatic stability of short peptides, the retroinverso analogue was prepared and tested for inhibitory activity of FIV in the presence or absence of normal cat serum. Differently from the unmodified peptide, the retroinverso analogue maintains strong inhibitory activity in serum. NMR studies showed that it displays crucial conformational features believed to be important for antiviral activity. PMID- 12723945 TI - Design, synthesis, and evaluation of benzothiadiazepine hydroxamates as selective tumor necrosis factor-alpha converting enzyme inhibitors. AB - Elevated levels of tumor necrosis factor-alpha (TNF-alpha) have been associated with several inflammatory diseases, and therefore, strategies for its suppression have become important targets in drug discovery. Our efforts to suppress TNF alpha have centered on the inhibition of TNF-alpha converting enzyme (TACE) through the use of hydroxamate inhibitors. Starting from broad-spectrum matrix metalloproteinase (MMP) inhibitors, we have designed and synthesized novel benzothiadiazepines as potent and selective TACE inhibitors. The benzothiadiazepines were synthesized with variation in P1 and P1' in order to effect potency and selectivity. The inhibitors were evaluated versus porcine TACE (pTACE), and the initial selectivity was assessed with counterscreens of MMP-1, 2, and -9. Several potent and selective inhibitors were discovered with compound 41 being the most active against pTACE (K(i) = 5 nM) while still maintaining good selectivity versus the MMP's (at least 75-fold). Most compounds were assessed in the human peripheral blood mononuclear cell assay (PBMC) and the human whole blood assay (WBA) to determine their ability to suppress TNF-alpha. Compound 32 was the most potent compound in the PBMC assay (IC(50) = 0.35 microM), while compound 62 was the most active in the WBA (IC(50) = 1.4 microM). PMID- 12723946 TI - Novel pyrazolo[3,4-d]pyrimidine-based inhibitors of Staphlococcus aureus DNA polymerase III: design, synthesis, and biological evaluation. AB - 6-Anilinopyrazolo[3,4-d]pyrimidin-4-ones are novel dGTP analogues that inhibit the replication-specific enzyme DNA polymerase III (DNA pol III) of Staphlococcus aureus and other Gram-positive (Gr+) bacteria. To enhance the potential of these inhibitors as antimicrobial agents, a structure-activity relationship was developed involving substitutions at the 2, 4, and pyrazolo NH positions. All of the new inhibitors were tested for their ability to inhibit S. aureus DNA pol III and the growth of several other Gr+ bacteria in culture. 2-Anilino groups with small hydrophobic groups in the meta or para position enhanced both antipolymerase and antimicrobial activity. 2-Benzyl-substituted inhibitors were substantially less active. Substitution in the 4-position by oxygen gave the optimal activity, whereas substitution at the pyrazolo NH was not tolerated. These pyrazolo[3,4-d]pyrimidine derivatives represent a novel class of antimicrobials with promising activities against Gr+ bacteria. PMID- 12723947 TI - Design, synthesis, and biological evaluation of monopyrrolinone-based HIV-1 protease inhibitors. AB - The design, synthesis, and biological evaluation of a series of HIV-1 protease inhibitors [(-)-6, (-)-7, (-)-23, (+)-24] based upon the 3,5,5-trisubstituted pyrrolin-4-one scaffold is described. Use of a monopyrrolinone scaffold leads to inhibitors with improved cellular transport properties relative to the earlier inhibitors based on bispyrrolinones and their peptide counterparts. The most potent inhibitor (-)-7 displayed 13% oral bioavailability in dogs. X-ray structure analysis of the monopyrrolinone compounds cocrystallized with the wild type HIV-1 protease provided valuable information on the interactions between the inhibitors and the HIV-1 enzyme. In each case, the inhibitors assumed similar orientations for the P2'-P1 substituents, along with an unexpected hydrogen bond of the pyrrolinone NH with Asp225. Interactions with the S2 pocket, however, were not optimal, as illustrated by the inclusion of a water molecule in two of the three inhibitor-enzyme complexes. Efforts to increase affinity by displacing the water molecule with second and third generation inhibitors did not prove successful. Lack of success with this venture is a testament to the difficulty of accurately predicting the many variables that influence and build binding affinity. Comparison of the inhibitor positions in three complexes with that of Indinavir revealed displacements of the protease backbones in the enzyme flap region, accompanied by variations in hydrogen bonding to accommodate the monopyrrolinone ring. The binding orientation of the pyrrolinone-based inhibitors may explain their sustained efficacy against mutant strains of the HIV-1 protease enzyme as compared to Indinavir. PMID- 12723948 TI - Rational design, synthesis, and structure-activity relationships of novel factor Xa inhibitors: (2-substituted-4-amidinophenyl)pyruvic and -propionic acids. AB - An inhibitor of factor Xa (fXa), the m-substituted benzamidine AXC1578 (1a), was structurally modified with the aim of increasing its potency. In particular, pyruvic acid and propionic acid substituents were incorporated into the P1 benzamidine moiety to introduce a favorable interaction with the oxy-anion hole in the catalytic triad region of fXa. This strategy was based on computational docking studies using the extracted active site of fXa. The validity of the computational model was supported by the acquisition of X-ray crystal structures of the 1a-trypsin and 3b-trypsin complexes (the homology around the active sites of fXa and trypsin is high). The above modifications significantly increased the inhibitory activity toward fXa, whereas the high selectivity for fXa versus thrombin was maintained or enhanced. Compounds 3b, 3c, 3e, and 4b are considered to be potential lead compounds for the development of orally active anticoagulant drugs because they demonstrated potent activity when administered orally to cynomolgus monkeys. PMID- 12723950 TI - 2-Amino-3-benzoylthiophene allosteric enhancers of A1 adenosine agonist binding: new 3, 4-, and 5-modifications. AB - 2-Amino-3-aroylthiophenes are agonist allosteric enhancers (AE) at the A(1) adenosine receptor (A(1)AR). Here we report the syntheses of three kinds of novel 2-aminothiophenes and assays of their AE activity at the human A(1)AR (hA(1)AR), namely, (1) 2-amino-4,5-diphenylthiophene-3-carboxylates, 3a-h, (2) 2-amino-3 benzoyl-4,5-diphenylthiophenes, 7a-p, and (3) 2-amino-5-bromo-3-benzoyl-4 phenylthiophenes, 10a-h. An in vitro assay employing the A(1)AR agonist [(125)I]ABA and membranes from CHO-K1 cells stably expressing the hA(1)AR measured an index of AE activity, the ability of a candidate AE to stabilize the agonist-A(1)AR-G protein ternary complex, scored as the percentage of ternary complex remaining after 10 min of dissociation initiated by CPX and GTPgammaS. The AE activity score of 2-amino-4,5-dimethyl-3-(3 trifluoromethylbenzoyl)thiophene (PD 81,723), which was 19%, served as a standard for comparison. Two 3-carboxythiophene 3-trifluoromethylbenzyl esters, 3d (49%) and 3f (63%), had substantial AE activity. The 3-(1-naphthoyl) substituent of 7e (52%) also supported AE activity. Compounds in series 3 tended to be more potent, 10a and 10c having scores of 91 and 80%, respectively. The activity of 2-amino-5 bromo-3-ethoxycarbonyl-4-(3-nitrophenyl)thiophene, 10h (26%), is an exception to the rule that a 3-ethoxycarbonyl substituent cannot support AE activity. PMID- 12723951 TI - Antiviral amphipathic oligo- and polyribonucleotides: analogue development and biological studies. AB - A series of novel N1 alkylated purine nucleic acids were polymerized either enzymatically or by automated synthesis to further establish the SAR requirements for HIV, RT, and HCMV activity. Out of the series, two constructs, 2'-O-methyl-1 allylinosinic acid phosphorothioate 33-mer (16) and an oligomer incorporating 1 propyl-6-thioinosinic acid residues (20), were found to be highly active under all three assay conditions. SAR studies indicate that sulfur incorporation, high molecular weight, and low steric bulk at N1 all can be important for activity. PMID- 12723949 TI - Effects of heterocyclic aromatic substituents on binding affinities at two distinct sites of somatostatin receptors. Correlation with the electrostatic potential of the substituents. AB - In our continuing program exploring glucose-based peptidomimetics of somatostatin (SRIF-14), we sought to improve the water solubility of our glycosides. This led to insights into the nature of the ligand binding sites at the SRIF receptor. Replacement of the C4 benzyl substituent in glucoside (+)-2 with pyridinylmethyl or pyrazin-2-ylmethyl congeners increased water solubility and enhanced affinity for the human SRIF subtype receptor 4 (sst4). We attribute this effect to hydrogen bond formation. The pyridin-3-ylmethyl substituent at C4, when combined with the imidazol-4-ylmethyl group at C2, generated (-)-19, which has the highest affinity of a glucose-based peptidomimetic at a human SRIF receptor to date (K(i) 53 +/- 23 nM, n = 6 at sst4). The C4 heterocyclic congeners of glucosides bearing a 1-methoxy substituent rather than an indole side chain at the anomeric carbon, such as (+)-16, also provided information about the Trp(8) binding pocket. We correlated the SARs at both the C4 and the Trp(8) binding pockets with calculations of the electrostatic potentials of the diverse C4 aromatic substituents using Spartan 3-21G(*) MO analysis. These calculations provide an approximate analysis of a molecule's ability to interact within a receptor binding site. Our binding studies show that benzene and indole rings, but not pyridinylmethyl nor pyrazin-2-ylmethyl rings, can bind the hydrophobic Trp(8) binding pocket of sst4. The Spartan 3-21G(*) MO analysis reveals significant negative electrostatic potential in the region of the pi-clouds for the benzene and indole rings but not for the pyridinylmethyl or pyrazin-2-ylmethyl congeners. Our data further demonstrate that the replacement of benzene or indole side chains by heterocyclic aromatic rings typified by pyridine and pyrazine not only enhances water solubility and hydrogen bonding capacity as expected, but can also profoundly diminish the ability of the pi-cloud of the aromatic substituent to interact with side chains of an aromatic binding pocket such as that for Trp(8) of SRIF-14. Conversely, these calculations accommodate the experimental findings that pyrazin-2-ylmethyl and pyridinylmethyl substituents at C4- of C1-indole substituted glycosides afford higher affinities at sst4 than the C4-benzyl group of (+)-2. This result is consistent with the high electron density in the plane of the heterocycle depicted in Figure 6 which can accept hydrogen bonds from the C4 binding pocket of the receptor. Unexpectedly, we found that the 2 fluoropyridin-5-ylmethyl analogue (+)-14 more closely resembles the binding affinity of (+)-8 than that of (+)-2, thus suggesting that (+)-14 represents a rare example of a carbon linked fluorine atom acting as a hydrogen bond acceptor. We attribute this result to the ability of the proton to bind the nitrogen and fluorine atoms simultaneously in a bifurcated arrangement. At the NK1 receptor of substance P (SP), the free hydroxyl at C4 optimizes affinity. PMID- 12723952 TI - Synthesis and evaluation of B-, C-, and D-ring-substituted estradiol carboxylic acid esters as locally active estrogens. AB - We have synthesized derivatives of estradiol that are structurally modified to serve as "soft" estrogens and act within a geographically limited area of the body; estrogens without systemic action. We have previously shown with 16alpha substituted analogues of estradiol that carboxylates proximal to the steroid ring neither bind to the estrogen receptor nor activate estrogen-responsive genes. However, when the carboxylic acid is masked as an ester, they bind to the receptor and stimulate estrogenic responses. Enzymatic hydrolysis through nonspecific esterases can inactivate these estrogens and thereby limit their area of action. Here, we describe our continued studies to design "soft" estrogens by synthesizing carboxylic acid esters of estradiol at the 7alpha-, 11beta-, and 15alpha-positions in the steroid nucleus at which bulky substituents are accommodated by the estrogen receptor. These compounds were tested for estrogen receptor binding (estrogen receptors alpha and beta), stimulation of an estrogen sensitive gene in Ishikawa cells in culture, and as substrates for enzymatic hydrolysis. Likely candidates were tested in in vivo assays for systemic and local estrogenic action. The biological studies showed that regardless of the point of attachment, all of the short-chain carboxylic acids, C-1 to C-3, were devoid of hormonal action, while many of the esters were estrogenic. The site on the steroid nucleus had great influence on hormonal activity and esterase hydrolysis. Formate esters at 7alpha and 15alpha were good estrogens, but lengthening the chain to acetate dramatically decreased hormonal activity. However, the 7alpha-formate esters were not enzymatically hydrolyzed. At 11beta, the acetate (methyl ester) was an effective estrogen, but increasing the chain length to propionate dramatically reduced hormonal activity. In general, the length of the alcohol from methyl to butyl had only a small effect on receptor binding, and as the size of the alcohol increased, so did esterase hydrolysis. One exception was the 11beta-acetate esters where increasing the alcohol moiety from methyl to ethyl eliminated estrogenic activity (Ishikawa cells) without affecting estrogen receptor binding. Several of the esters were tested in vivo, and two, the methyl and ethyl esters of estradiol-15alpha-formate, appeared to have the requisite properties (high local and low systemic activity) of superior "soft" estrogens. PMID- 12723953 TI - A novel antibacterial 8-chloroquinolone with a distorted orientation of the N1-(5 amino-2,4-difluorophenyl) group. AB - Fluoroquinolones represent a major class of antibacterial agents with great therapeutic potential. In this study, we designed m-aminophenyl groups as novel N 1 substituents of naphthyridones and quinolones. Among newly synthesized compounds, 7-(3-aminoazetidin-1-yl)-1-(5-amino-2,4-difluorophenyl)-8-chloro-6 fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid (4) has extremely potent antibacterial activities against Gram (+) as well as Gram (-) bacteria. This compound is significantly more potent than trovafloxacin against clinical isolates: 30 times against Streptococcus pneumoniae and 128 times against methicillin resistant Staphylococcus aureus. The structure-activity relationship (SAR) study revealed that a limited combination of 1-(5-amino-2,4-difluorophenyl) group, 7-(azetidin-1-yl) group, and 8-Cl atom (or Br atom or Me group) gave potent antibacterial activity. An X-ray crystallographic study of a 7-(3 ethylaminoazetidin-1-yl)-8-chloro derivative demonstrated that the N-1 aromatic group was remarkably distorted out of the core quinolone plane by steric repulsion between the C-8 Cl atom and the N-1 substituent. Furthermore, a molecular modeling study of 4 and its analogues demonstrated that a highly distorted orientation was induced by a steric hindrance of the C-8 substituent, such as Cl, Br, or a methyl group. Thus, their highly strained conformation should be a key factor for the potent antibacterial activity. PMID- 12723955 TI - Inhibition of papilloma formation by analogues of 7,8-dihydroretinoic acid. AB - The design of analogues of 7,8-dihydroretinoic acid (7,8-dihydro-RA) was based on reported biological activities of this retinoid and its dihydro-TMMP(1) analogue and on structural hypotheses. 7-Oxa-7,8-dihydroretinoids (5, 6) were prepared by O-alkylation of phenoxides by methyl 8-bromo-3,7-dimethyl-2,4,6-octatrienoate. In some cases, C-alkylation also occurred. 7-Aza-8-oxo-7,8-dihydroretinoids (12, 13) were synthesized from benzeneamines and the acyl cyano or bromo derivative of the monomethyl ester of 3,7-dimethyl-2,4,6-octatriene-1,8-dioic acid. These monomethyl ester precursors were synthesized from the known analogous aldehyde via an O-trimethylsilyl cyanohydrin. 7-(2,3,5-Trimethylphenoxy)-3,5-dimethyl 2,4,6-octatrienoic acid (6b) was the most active of the 7-oxa-7,8-dihydro-RAs in inhibiting DMBA-initiated and TPA-promoted mouse-skin papillomas. The ED(50) was about 4-fold that of etretinate. Two additional 7-oxa-7,8-dihydro-RAs exhibited modest activity in the papilloma assay. Some of the 7-oxa-7,8-dihydro-RAs bind to CRABP and RARalpha. PMID- 12723954 TI - Design of selective peptidomimetic agonists for the human orphan receptor BRS-3. AB - New tool substances may help to unravel the physiological role of the human orphan receptor BRS-3 and its possible use as a drug target for the treatment of obesity and cancer. In continuation of our work on BRS-3, the solid- and solution phase synthesis of a library of low molecular weight peptidomimetic agonists based on the recently developed short peptide agonist 4 is described. Functional potencies of the compounds were determined measuring calcium mobilization in a fluorometric imaging plate reader (FLIPR) assay. Focusing on the N-terminus, the d-Phe-Gln moiety of 4 was modified in a combinatorial SAR-oriented medicinal chemistry approach. With the incorporation of N-arylated glycine and alanine building blocks azaglycine, piperazine, or piperidine and the synthesis of semicarbazides and semicarbazones, a number of highly potent and selective compounds with a reduced number of peptide bonds were obtained, which also should have enhanced metabolic stability. PMID- 12723956 TI - Molecular modeling calculations of HIV-1 reverse transcriptase nonnucleoside inhibitors: correlation of binding energy with biological activity for novel 2 aryl-substituted benzimidazole analogues. AB - The energies and physical descriptors for the binding of 20 novel 1-(2,6 difluorobenzyl)-2-(2,6-difluorophenyl)benzimidazole analogues (BPBIs) to HIV-1 reverse transcriptase (RT) have been determined using Monte Carlo (MC) simulations. The crystallographic structure of the lead compound, 1-(2,6 difluorobenzyl)-2-(2,6-difluorophenyl)-4-methylbenzimidazole, was used as a starting point to model the inhibitors in both the bound and the unbound states. The energy terms and physical descriptors obtained from the calculations were correlated with their respective experimental EC(50) values, resulting in an r(2) value of 0.70 and a root-mean-square deviation (rms) of 0.53 kcal/mol. The terms in the correlation include the change in total Coulombic energy and solvent accessible surface area. Structural analysis of the data files from the BPBI calculations reveals that all of the analogues with good biological activity show the formation of a hydrogen bond between the ligand and the backbone nitrogen atom of lysine 103. By use of the structural results, two novel BPBI inhibitors have been designed and calculations have been carried out. The results show the formation of the desired hydrogen bonds, and the DeltaG(binding) values predict the compounds to be excellent RT inhibitors. Subsequent synthesis and biological activity testing of these analogues have shown the validity of the predictive calculations. If the BPBIs are modeled in a site constructed from the crystal coordinates of a member of another class of nonnucleoside inhibitors (the 4,5,6,7 tetrahydroimidazo[4,5,1-jk][1,4]benzodiazepine-2(1H)-thione and -one (TIBO) compounds), the correlation with the same terms drops slightly, giving an r(2) value of 0.61 with an associated root-mean-square value of 0.53 kcal/mol. Conversely, if the TIBO compounds are modeled in a site constructed from the BPBI complex crystal coordinates, a correlation can be obtained using the drug-protein interaction energy and change in the total number of hydrogen bonds, giving an r(2) value of 0.63. These are the same descriptors that were used for the TIBO compounds modeled in their own sites, where the r(2) value was 0.72. These data suggest that it may be possible, in some cases, to design novel inhibitors utilizing structural data from related, but not identical, inhibitors. PMID- 12723958 TI - Antimalarial activity of novel 1,2,5,6-tetraoxacycloalkanes and 1,2,5 trioxacycloalkanes. AB - Photooxygenation of 2-phenylnorbornene 1 in the presence of 30% aqueous hydrogen peroxide afforded 1,2-bishydroperoxide 3, which could be cycloalkylated on treatment with silver oxide and a 1,omega-diiodoalkane to provide the tricyclic peroxides 12. Trimethylsilylation of 3 followed by TMSOTf-catalyzed cyclocondensation with carbonyl compounds led to the formation of the tricyclic peroxides 14 containing a 1,2,4,5-tetroxepane structure. Photooxygenation of 1 in the presence of either unsaturated hydroperoxides or unsaturated alcohols followed by bis(collidine)iodine hexafluorophosphate promoted cyclization gave the corresponding cyclic peroxides 15-17. Several of these cyclic peroxides showed substantial antimalarial activity particularly in vitro. PMID- 12723957 TI - Structural properties of dibenzosuberanylpiperazine derivatives for efficient reversal of chloroquine resistance in Plasmodium chabaudi. AB - For the purpose of developing chemosensitizers to reverse chloroquine (CQ) resistance in Plasmodium chabaudi in vivo, dibenzosuberanylpiperazine (1-(10,11 dihydro-5H-dibenzo[a,d]cyclohepten-5-yl)piperazine) (DSP) and its piperazin-1-yl derivatives were synthesized systematically. DSP hydrochloride (3) was obtained from the reaction of dibenzosuberanyl chloride with piperazine in the presence of 1,8-diazabicyclo[5,4,0]-7-undecene (DBU). To understand the relationship between the substituent patterns of DSP derivatives and their biological activities, 13 hydroxyalkyl or hydroxyalkenyl derivatives were synthesized by an attack of the piperazine secondary amine of 3 on commercially available epoxides in the presence of triethylamine or DBU, and three alkyl or alkynyl derivatives were synthesized by the reactions of 3 with the corresponding organic chlorides in the presence of DBU. In both reactions, the yield was a maximum of 90%. The biological activities of the synthesized compounds were evaluated on the basis of two values: antimalarial activity and reversal activity. The values of antimalarial activities by single administration of 17 test compounds were not effective, being in the range 67-152% on day 4 after infection of Plasmodium chabaudi to mice except for the administration of 3-(dibenzosuberanylpiperazin-1 yl)-1-butene (29, 22%). On the other hand, administration of the seven test compounds (50 mg/kg dose) combined with CQ (3-4 mg/kg) gave high reversal activities, namely, low values (0% on day 4). The effective test compounds were those obtained by introducing the following substituents: 2-hydroxybutyl (24), 2 hydroxyhexen-5-yl (27), 2-hydroxybuten-3-yl (28a), 2-substituted 1-hydroxybuten-3 yl (28b), 4-acetoxybutyn-2-yl (30), 4-hydroxybutyn-2-yl (31), and 3-substituted buten-1-yl (29), which correspond to the nonbulky groups of hydroxyalkyl (C4), hydroxyalkenyl (C4-C6), hydroxyalkynyl (C4), or alkenyl (C4). These results may lead to the development of an approach to developing clinically applicable chemosensitizers for drug-resistant malaria. PMID- 12723959 TI - Synthesis and biological evaluation of new 2-(4,5-dihydro-1H-imidazol-2-yl)-3,4 dihydro-2H-1,4-benzoxazine derivatives. AB - 2-(4,5-Dihydro-1H-imidazol-2-yl)-3,4-dihydro-2H-1,4-benzoxazine derivatives and tricyclic analogues with a fused additional ring on the nitrogen atom of the benzoxazine moiety have been prepared and evaluated for their cardiovascular effects as potential antihypertensive agents. The imidazoline ring was generated by reaction of the corresponding ethyl ester with ethylenediamine. Affinities for imidazoline binding sites (IBS) I(1) and I(2) and alpha(1) and alpha(2) adrenergic receptors were evaluated as well as the effects on mean arterial blood pressure (MAP) and heart rate (HR) of spontaneously hypertensive rats. With few exceptions the most active compounds on MAP were those with high affinities for IBS and alpha(2) receptor. Among these, compound 4h was the most interesting and is now, together with its enantiomers, under complementary pharmacological evaluation. PMID- 12723960 TI - Cyclopropane-based conformational restriction of histamine. (1S,2S)-2-(2 aminoethyl)-1-(1H-imidazol-4-yl)cyclopropane, a highly selective agonist for the histamine H3 receptor, having a cis-cyclopropane structure. AB - A series of cyclopropane-based conformationally restricted analogues of histamine, the "folded" cis-analogues, i.e., (1S,2R)-2-(aminomethyl)-1-(1H imidazol-4-yl)cyclopropane (11), (1S,2S)-2-(2-aminoethyl)-1-(1H-imidazol-4 yl)cyclopropane (13), and their enantiomers ent-11 and ent-13, and the "extended" trans-analogues, i.e., (1R,2R)-2-(aminomethyl)-1-(1H-imidazol-4-yl)cyclopropane (12) and its enantiomer ent-12, were designed as histamine H(3) receptor agonists. These target compounds were synthesized from the versatile chiral cyclopropane units, (1S,2R)- and (1R,2R)-2-(tert-butyldiphenylsilyloxy)methyl-1 formylcyclopropane (14 and 15, respectively) or their enantiomers ent-14 and ent 15. Among the conformationally restricted analogues, the "folded" analogue 13 (AEIC) having the cis-cyclopropane structure was identified as a potent H(3) receptor agonist, which showed a significant binding affinity (K(i) = 1.31 +/- 0.16 nM) and had an agonist effect (EC(50) value of 10 +/- 3 nM) on the receptor. This compound owes its importance to being the first highly selective H(3) receptor agonist to have virtually no effect on the H(4) subtype receptor. These studies showed that the cis-cyclopropane structure is very effective in the conformational restriction of histamine to improve the specific binding to the histamine H(3) receptor. PMID- 12723962 TI - Biaryl analogues of conformationally constrained tricyclic tropanes as potent and selective norepinephrine reuptake inhibitors: synthesis and evaluation of their uptake inhibition at monoamine transporter sites. AB - A series of novel conformationally constrained tricyclic tropane derivatives containing a biaryl moiety, (Z)-9-(biarylylmethylene)-7 azatricyclo[4.3.1.0(3,7)]decanes, were synthesized and evaluated for their ability to inhibit reuptake of dopamine (DA), serotonin (5-HT), and norepinephrine (NE) by the DA, 5-HT, and NE transporters. Most of the compounds containing a methoxycarbonyl substituent at C-10 exhibit moderate to high inhibitory activity at the NET but lower activity at the DAT and SERT. Among these new compounds, some potent, NET-selective ligands were identified. The p methoxy derivative 11a has a K(i) value of 39 nM for uptake inhibition at the NET and moderate to high selectivity over the SERT (100-fold) and the DAT (20-fold). Compound 11f exhibits a remarkable potency (K(i) = 9.7 nM) at the NET and a 25 fold selectivity over both the SERT and the DAT. Analogue 23 containing a thiophene ring as a bioisosteric replacement of the phenyl ring Ar(1) displays a high activity (K(i) = 10.3 nM) for the NET and similar selectivity over the SERT (50-fold) and the DAT (37-fold). The selectivity profile of biaryl analogues differs from that of the monoaryl series, as most members of that series display excellent potency at and selectivity for the SERT (J. Med. Chem. 2002, 45, 1930). This finding suggests that the different shape and size of the lipophilic recognition pocket that encompasses the aryl ring(s) of these tropanes are major determinants of a ligand's transporter activity at either the NET or the SERT. Some of the compounds in this series may also be valuable in sorting out the contribution of the individual transporters to cocaine's reinforcing properties. PMID- 12723961 TI - Synthesis and biological evaluation of thiol-based inhibitors of glutamate carboxypeptidase II: discovery of an orally active GCP II inhibitor. AB - A series of 2-(thioalkyl)pentanedioic acids were synthesized and evaluated as inhibitors of glutamate carboxypeptidase II (GCP II, EC 3.4.17.21). The inhibitory potency of these thiol-based compounds against GCP II was found to be dependent on the number of methylene units between the thiol group and pentanedioic acid. A comparison of the SAR of the thiol-based inhibitors to that of the phosphonate-based inhibitors provides insight into the role of each of the two zinc-binding groups in GCP II inhibition. The most potent thiol-based inhibitor, 2-(3-mercaptopropyl)pentanedioic acid (IC(50) = 90 nM), was found to be orally bioavailable in rats and exhibited efficacy in an animal model of neuropathic pain following oral administration. PMID- 12723964 TI - Extrinsic cues aid shape recognition from novel viewpoints. AB - It has been shown previously that the visual recognition of shape is susceptible to the mismatch between the retinal input and its representation in long-term memory, especially when this mismatch arises from rotations in depth. One possibility is that the visual recognition system deals with such mismatch by a transformation of the input or the representation thereby bringing both into alignment for comparison. In either case, knowing what transformation has taken place should facilitate recognition. In natural circumstances, objects do not disappear and appear in different orientations inexplicably and an observer usually knows what to expect according to the context. This context includes the environment, and the history of the observers' movements, which specify the transient relationship between the object, the environment and the observer. We used interactive computer graphics to study the effects of providing observers with either implicit or explicit indications of their view transformations in the recognition of a class of shape found previously to be highly view-dependent. Results show that these cues aid recognition to varying degrees but mostly for oblique views and primarily in terms of accuracy not response times. These results provide evidence for egocentric encoding of shape and suggest that knowing ones' transformation in view helps to reduce the problem space involved in matching a shape percept with a mental representation. PMID- 12723965 TI - The detection of colored Glass patterns. AB - The detection of many chromatic stimuli is mediated by mechanisms that sum their inputs linearly. As a result, these mechanisms have a broad range of selectivity in color space, as do the majority of cells in the early stages of visual processing. In extrastriate cortex, there are cells with a narrow tuning in color space. The function of these cells is not fully understood: they could be involved in color categorization, or could mediate the detection of stimuli such as Glass patterns, whose properties make them undetectable by early stages of processing. We measured the tuning properties of the mechanisms responsible for the detection of colored Glass patterns and found that they have a broad tuning in color space. Our results suggest that Glass patterns are detected by a multitude of mechanisms that sum their inputs linearly. PMID- 12723963 TI - Synthesis and structure-activity relationships of cis tetrahydrophthalazinone/pyridazinone hybrids: a novel series of potent dual PDE3/PDE4 inhibitory agents. AB - In this study, the synthesis and in vitro and in vivo pharmacological investigations of a new series of phthalazinone/pyridazinone hybrids with both PDE3 and PDE4 inhibitory activities are described. These compounds combine the pharmacophores of recently discovered 4a,5,8,8a-tetrahydro-2H-phthalazin-1-one type inhibitors of PDE4 and the well-known 2H-pyridazin-3-one-type PDE3 inhibitors such as the tetrahydrobenzimidazoles. Most of the synthesized compounds are pharmacologically spoken PDE3/PDE4 hybrids. All hybrids show potent PDE4 inhibitory activity (pIC(50) = 7.0-8.7), whereas the pIC(50) values for inhibition of PDE3 vary from 5.4 to 7.5. In general, analogues with a 5-methyl 4,5-dihydropyridazinone moiety exhibit the highest PDE3 inhibitory activities. The highest in vivo antiinflammatory activity is displayed by phthalazinones 43 and 44 showing, at a dose of 30 micromol/kg po, 46% inhibition of arachidonic acid (AA) induced mouse ear edema. No correlation was found between the in vitro PDE3 and/or PDE4 inhibitory activity and the in vivo antiinflammatory capacity after oral dosing. PMID- 12723966 TI - Comparison of two weighted integration models for the cueing task: linear and likelihood. AB - In a task in which the observer must detect a signal at two locations, presenting a precue that predicts the location of a signal leads to improved performance with a valid cue (signal location matches the cue), compared to an invalid cue (signal location does not match the cue). The cue validity effect has often been explained with a limited capacity attentional mechanism improving the perceptual quality at the cued location. Alternatively, the cueing effect can also be explained by unlimited capacity models that assume a weighted combination of noisy responses across the two locations. We compare two weighted integration models, a linear model and a sum of weighted likelihoods model based on a Bayesian observer. While qualitatively these models are similar, quantitatively they predict different cue validity effects as the signal-to-noise ratios (SNR) increase. To test these models, 3 observers performed in a cued discrimination task of Gaussian targets with an 80% valid precue across a broad range of SNR's. Analysis of a limited capacity attentional switching model was also included and rejected. The sum of weighted likelihoods model best described the psychophysical results, suggesting that human observers approximate a weighted combination of likelihoods, and not a weighted linear combination. PMID- 12723967 TI - The origin of the oblique effect examined with pattern adaptation and masking. AB - The decreased visibility of obliquely oriented patterns as compared to horizontal or vertical ones is termed the oblique effect. The origin of the oblique effect in the chain of visual processing was examined by comparing the potency of oblique adapting gratings to the potency of horizontal ones. Oblique gratings (which were less visible but of equal physical contrast) were as powerful or more powerful than horizontal gratings as adapting stimuli. Obliquely oriented stimuli also produced a slightly stronger tilt aftereffect than stimuli near the cardinal axes. These results suggest that the diminished neural representation of oblique stimuli arises in the human cortex, rather than from impairments of sensitivity or resolution in the initial geniculo-cortical projection. PMID- 12723968 TI - Extraocular connective tissue architecture. AB - Extraocular muscle pulleys, now well known to be kinematically significant extraocular structures, have been noted in passing and described in fragments several times over the past two centuries. They were late to be fully appreciated because biomechanical modeling of the orbit was not available to derive their kinematic consequences, and because pulleys are distributed condensations of collagen, elastin and smooth muscle (SM) that are not sharply delineated. Might other mechanically significant distributed extraocular structures still be awaiting description?An imaging approach is useful for describing distributed structures, but does not seem suitable for assessing mechanical properties. However, an image that distinguished types and densities of constituent tissues could give strong hints about mechanical properties. Thus, we have developed methods for producing three dimensional (3D) images of extraocular tissues based on thin histochemically processed slices, which distinguish collagen, elastin, striated muscle and SM. Overall tissue distortions caused by embedding for sectioning, and individual-slice distortions caused by thin sectioning and subsequent histologic processing were corrected by ordered image warping with intrinsic fiducials. We describe an extraocular structure, partly included in Lockwood's ligament, which contains dense elastin and SM bands, and which might refine horizontal eye alignment as a function of vertical gaze, and torsion in down-gaze. This active structure might therefore be a factor in strabismus and a target of therapeutic intervention. PMID- 12723969 TI - Inhibition of liver trans-sulphuration pathway by propargylglycine mimics gene expression changes found in the mammary gland of weaned lactating rats: role of glutathione. AB - In the lactating mammary gland, weaning produces mitochondrial cytochrome c release and nuclear DNA fragmentation, as determined by gel electrophoresis. This is followed by a significant decrease in lactation. Weaning for 2 h produces an early induction of the tumour suppressor/transcription factor p53, whereas the oncoprotein c-Jun and c-Jun N-terminal kinase are elevated after 24 h of weaning when compared with controls. The expression of p21(cip1) and p27(kip1), cyclin dependent kinase inhibitors, was significantly higher in weaned rats when compared with control lactating rats. All the changes mentioned above also happen in the lactating mammary gland when propargylglycine, an inhibitor of the liver trans-sulphuration pathway, is administered. This effect is partially reversed by N -acetylcysteine administration. The administration of buthionine sulphoximine, an irreversible inhibitor of gamma-glutamylcysteine synthetase, to lactating rats produces a decrease in GSH levels and changes in protein concentrations and gene transcripts similar to those in rats with impaired trans-sulphuration pathway. These data suggest that the inter-tissue flux of GSH is an important mechanism of L-cysteine delivery to the lactating mammary gland and emphasize the importance of this physiological event in maintaining the gene expression required to sustain lactation. PMID- 12723970 TI - Free-oligosaccharide control in the yeast Saccharomyces cerevisiae: roles for peptide:N-glycanase (Png1p) and vacuolar mannosidase (Ams1p). AB - Free oligosaccharides (fOS) are generated during glycoprotein biosynthesis in mammalian cells. Here we report on the origin and fate of these structures in the yeast Saccharomyces cerevisiae. After metabolic radiolabelling with [2 (3)H]mannose ([2-(3)H]Man) for 30 min, Man(8)GlcNAc(2) was identified as the predominant fOS in this organism, and radioactivity associated with this structure was found to correspond to approximately 1% of that associated with the same structure N -linked to glycoprotein. Despite provoking a fourfold increase in radioactivity associated with lipid-linked oligosaccharide, the protein synthesis inhibitor cycloheximide blocked [2-(3)H]Man incorporation into both endo-beta-D- N -acetylglucosamine H-sensitive N-glycans and fOS. Peptide:N glycanase, encoded by the PNG1 gene, was found to be required for the generation of a large proportion of yeast fOS during, and soon after, protein glycosylation. Use of an ams1 Delta strain deficient in the vacuolar alpha-mannosidase revealed this enzyme to be responsible for the slow growth-associated catabolism of fOS. The present paper constitutes the first description of fOS formation in intact S. cerevisiae, and, with the demonstration that fOS are degraded by the vacuolar mannosidase, a novel function for this poorly understood enzyme has been identified. PMID- 12723972 TI - On the substrate specificity of bacterial DD-peptidases: evidence from two series of peptidoglycan-mimetic peptides. AB - The reactions between bacterial DD-peptidases and beta-lactam antibiotics have been studied for many years. Less well understood are the interactions between these enzymes and their natural substrates, presumably the peptide moieties of peptidoglycan. In general, remarkably little activity has previously been demonstrated in vitro against potential peptide substrates, although in many cases the peptides employed were non-specific and not homologous with the relevant peptidoglycan. In this paper, the specificity of a panel of DD peptidases against elements of species-specific D-alanyl-D-alanine peptides has been assessed. In two cases, those of soluble, low-molecular-mass DD-peptidases, high activity against the relevant peptides has been demonstrated. In these cases, the high specificity is towards the free N-terminus of the peptidoglycan fragment. With a number of other enzymes, particularly high-molecular-mass DD peptidases, little or no activity against these peptides was observed. In separate experiments, the reactivity of the enzymes against the central, largely invariant, peptide stem was examined. None of the enzymes surveyed showed high activity against this structural element although weak specificity in the expected direction towards the one structural variable (D-gammaGln versus D gammaGlu) was observed. The current state of understanding of the activity of these enzymes in vitro is discussed. PMID- 12723971 TI - Differential role of glutaredoxin and thioredoxin in metabolic oxidative stress induced activation of apoptosis signal-regulating kinase 1. AB - Redox-sensing molecules such as thioredoxin (TRX) and glutaredoxin (GRX) bind to apoptosis signal-regulating kinase 1 (ASK1) and suppress its activation. Glucose deprivation disrupted the interaction between TRX/GRX and ASK1 and subsequently activated the ASK1-stress-activated protein kinase/extracellular-signal-regulated kinase kinase-c-Jun N-terminal kinase 1 (JNK1) signal-transduction pathway. L Buthionine-( S, R )-sulphoximine, which decreases intracellular glutathione content, enhanced glucose deprivation-induced activation of JNK1 by promoting the dissociation of TRX, but not GRX, from ASK1. Treatment of cells with exogenous glutathione disulphide ester resulted in the dissociation of GRX, but not TRX, from ASK1 and the subsequent activation of JNK1. Nonetheless, overexpression of calatase, an H(2)O(2) scavenger, inhibited JNK1 activation and cytotoxicity as well as the dissociation of TRX and GRX from ASK1 during combined glucose deprivation and L-buthionine-( S, R )-sulphoximine treatment. Taken together, glucose deprivation-induced metabolic oxidative stress may activate ASK1 through two different pathways: glutathione-dependent GRX-ASK1 and glutathione independent TRX-ASK1 pathways. PMID- 12723973 TI - Complexes between the nonsense-mediated mRNA decay pathway factor human upf1 (up frameshift protein 1) and essential nonsense-mediated mRNA decay factors in HeLa cells. AB - mRNAs harbouring premature translation-termination codons are usually degraded by the nonsense-mediated mRNA decay (NMD) pathway. Human up-frameshift protein 1 (Hupf1) is an NMD factor that is conserved between yeast and mammals. To isolate cellular complexes that are formed with Hupf1 and to explore the role of cellular proteins in NMD, we generated a HeLa cell line that stably expresses Hupf1 bearing a double-affinity tag (termed Hupf1-2tag). Hupf1-2tag is localized in the cytoplasm similar to the endogenous Hupf1 protein, and the Hupf1-2tag cell line is fully NMD-competent. Using affinity chromatography, Hupf1-2tag-associated proteins were isolated. MS and immunoblotting identified the NMD factors Hupf2 and Hupf3a/b as interaction partners of Hupf1. Size-exclusion chromatography indicates that the NMD factors Hupf1, Hupf2 and the large isoform of Hupf3a might exist in a stable, high-molecular-mass complex of approx. 1.3 MDa. Interestingly, the poly(A)-binding protein was also identified by MS to be associated specifically with Hupf1-2tag. In contrast with the interaction with Hupf2 and Hupf3a/b, the association of poly(A)-binding protein with Hupf1 is highly sensitive to treatment of the isolated complexes with RNase. Components of the exon-exon junction complex or the translational eukaryotic release factor (eRF) 3 were not identified in complexes associated with Hupf1-2tag. We discuss these findings in the context of current models of NMD. PMID- 12723974 TI - Biochemical mechanism of action of a diketopiperazine inactivator of plasminogen activator inhibitor-1. AB - XR5118 [(3 Z,6 Z )-6-benzylidine-3-(5-(2-dimethylaminoethyl-thio-))-2 (thienyl)methylene-2,5-dipiperazinedione hydrochloride] can inactivate the anti proteolytic activity of the serpin plasminogen activator inhibitor-1 (PAI-1), a potential therapeutic target in cancer and cardiovascular diseases. Serpins inhibit their target proteases by the P(1) residue of their reactive centre loop (RCL) forming an ester bond with the active-site serine residue of the protease, followed by insertion of the RCL into the serpin's large central beta-sheet A. In the present study, we show that the RCL of XR5118-inactivated PAI-1 is inert to reaction with its target proteases and has a decreased susceptibility to non target proteases, in spite of a generally increased proteolytic susceptibility of specific peptide bonds elsewhere in PAI-1. The properties of XR5118-inactivated PAI-1 were different from those of the so-called latent form of PAI-1. Alanine substitution of several individual residues decreased the susceptibility of PAI-1 to XR5118. The localization of these residues in the three-dimensional structure of PAI-1 suggested that the XR5118-induced inactivating conformational change requires mobility of alpha-helix F, situated above beta-sheet A, and is in agreement with the hypothesis that XR5118 binds laterally to beta-sheet A. These results improve our understanding of the unique conformational flexibility of serpins and the biochemical basis for using PAI-1 as a therapeutic target. PMID- 12723976 TI - COX-2: where are we in 2003?--distinction from NSAIDs becoming blurred. AB - The distinction between cyclooxygenase-2-selective inhibitors (CSIs) and nonsteroidal anti-inflammatory drugs ultimately must be clinical and must be clinically and economically relevant. This distinction needs to be demonstrated in a substantial and clinically relevant difference in the respective rates of serious adverse reactions of the upper gastrointestinal tract. Event-driven, randomized, blinded, controlled trials with sufficient power are required to resolve uncertainties concerning the relative risk of thrombotic cardiovascular events in patients taking CSIs who have risk factors for these events. Patients and situations more representative of those in primary-care practice - elderly, comorbidities, comedication - need to be included in larger studies to provide a better understanding of the risks and benefits of CSIs. PMID- 12723975 TI - CD44 in rheumatoid arthritis. AB - CD44 is a multistructural cell-surface glycoprotein that can theoretically generate close to 800 isoforms by differential alternative splicing. At present, several dozen isoforms are known. The polymorphic nature of CD44 might explain its multifunctionality and its ability to interact with many cell-surface and extracellular ligands, the principal one being hyaluronic acid (HA). Of the many CD44 functions, our review focuses on its involvement in cell-cell and cell matrix interactions, as well as on its implication in the support of cell migration and the presentation of growth factors to their cognate receptors. Cells involved in pathological activities such as cancer cells and destructive inflammatory cells, and also normal cells engaged in physiological functions, use cell-surface CD44 for their localization and expansion at extravascular sites. This article reviews the evidence that the joint synovium of patients with rheumatoid arthritis (RA) contains considerable amounts of various CD44 isoforms as well as the HA ligand. The review also shows that anti-CD44 monoclonal antibody (mAb) directed against constant epitopes, shared by all CD44 isoforms, can markedly reduce the inflammatory activity of arthritis induced by collagen or proteoglycans in mice. Anti-CD44 mAb also interferes with the migration of RA synovial-like fibroblasts in vitro and is able to disturb the destructive interaction between RA synovial-like fibroblasts and the cartilaginous matrix. However, the transition from the experimental model to the patient's bedside is dependent on the ability to target the CD44 of cells engaged in RA pathology, while skipping the CD44 of normal cells. PMID- 12723978 TI - B cells as a therapeutic target in autoimmune disease. AB - Depleting B cells with anti-CD20 monoclonal antibodies emerges as a new therapeutic strategy in autoimmune diseases. Preliminary clinical studies suggest therapeutic benefits in patients with classic autoantibody-mediated syndromes, such as autoimmune cytopenias. Treatment responses in rheumatoid arthritis have opened the discussion about whether mechanisms beyond the removal of potentially pathogenic antibodies are effective in B-cell depletion. B cells may modulate T cell activity through capturing and presenting antigens or may participate in the neogenesis of lymphoid microstructures that amplify and deviate immune responses. Studies exploring which mechanisms are functional in which subset of patients hold the promise of providing new and rational treatment approaches for autoimmune syndromes. PMID- 12723977 TI - Degeneration of the intervertebral disc. AB - The intervertebral disc is a cartilaginous structure that resembles articular cartilage in its biochemistry, but morphologically it is clearly different. It shows degenerative and ageing changes earlier than does any other connective tissue in the body. It is believed to be important clinically because there is an association of disc degeneration with back pain. Current treatments are predominantly conservative or, less commonly, surgical; in many cases there is no clear diagnosis and therapy is considered inadequate. New developments, such as genetic and biological approaches, may allow better diagnosis and treatments in the future. PMID- 12723979 TI - SLE--systemic lupus erythematosus: a BLySful, yet BAFFling, disorder. AB - BLyS/BAFF (B-lymphocyte stimulator/B-cell activating factor) is a vital B-cell survival factor. Overexpression of BLyS in mice may lead to systemic-lupus erythematosus-like (SLE-like) disease, and BLyS overexpression is common in human SLE. Treatment of SLE-prone mice with a BLyS antagonist ameliorates disease progression and enhances survival, making BLyS an attractive therapeutic target in human disease. However, several unresolved issues remain, including what is the contributory role of APRIL (a tumor-necrosis-factor superfamily member related to BLyS) in the 'autoimmunogenic' effects of BLyS, identification of the 'optimal' BLyS antagonist, and identification of those SLE patients most likely to benefit from BLyS antagonist therapy. PMID- 12723980 TI - Behcet's disease. AB - Behcet's disease is characterized by recurrent aphthous stomatitis, uveitis, genital ulcers, and skin lesions. The role of the HLA-B*51 gene has been confirmed in recent years, although its contribution to the overall genetic susceptibility to Behcet's disease was estimated to be only 19%. The production of a variety of cytokines by T cells activated with multiple antigens has been shown to play a pivotal role in the activation of neutrophils. As regards the treatment, anti-tumor necrosis factor alpha therapy has been shown to be effective for mucocutaneous symptoms as well as for sight-threatening panuveitis, although a randomized, controlled trial is required. PMID- 12723981 TI - Autoantibodies in vasculitis. AB - Before the mid-1980s the only autoantibody widely used to assist in diagnosing vasculitic disease was IgG antibody to the alpha(3) domain of the noncollagenous part of type IV collagen (anti-glomerular basement membrane). Since that time, antineutrophil cytoplasmic antibodies (ANCAs) directed at the azurophilic granule proteins proteinase-3 and myeloperoxidase have been established as clinically useful autoantibodies to support a diagnosis of Wegener's granulomatosis, microscopic polyangiitis, Churg-Strauss syndrome and limited forms of these primary, small vessel necrotizing and often granulomatous vasculitides. The establishment of standardized methods for identifying those antibodies was needed before they could be used in clinical practice. The levels of both types of ANCAs tend to increase in parallel with the degree of clinical disease activity, and they decrease with successful immunosuppressive therapy. More than one assay may have to be used to discover imminent exacerbations in proteinase-3-ANCA associated syndromes. Although autoantibodies to endothelial cells may be important players in the pathogenesis of several vasculitic conditions, they have not gained clinical popularity because of lack of standardized detection methods. PMID- 12723983 TI - New insights into integrin signalling: implications for rheumatoid arthritis synovial fibroblasts. PMID- 12723985 TI - Adverse effects of adenovirus-mediated gene transfer of human transforming growth factor beta 1 into rabbit knees. AB - To examine the effect of transforming growth factor (TGF)-beta1 on the regulation of cartilage synthesis and other articular pathologies, we used adenovirus mediated intra-articular gene transfer of TGF-beta1 to both naive and arthritic rabbit knee joints. Increasing doses of adenoviral vector expressing TGF-beta1 were injected into normal and antigen-induced arthritis rabbit knee joints through the patellar tendon, with the same doses of an adenoviral vector expressing luciferase injected into the contralateral knees as the control. Intra articular injection of adenoviral vector expressing TGF-beta1 into the rabbit knee resulted in dose-dependent TGF-beta1 expression in the synovial fluid. Intra articular TGF-beta1 expression in both naive and arthritic rabbit knee joints resulted in significant pathological changes in the rabbit knee as well as in adjacent muscle tissue. The observed changes induced by elevated TGF-beta1 included inhibition of white blood cell infiltration, stimulation of glycosaminoglycan release and nitric oxide production, and induction of fibrogenesis and muscle edema. In addition, induction of chondrogenesis within the synovial lining was observed. These results suggest that even though TGF beta1 may have anti-inflammatory properties, it is unable to stimulate repair of damaged cartilage, even stimulating cartilage degradation. Gene transfer of TGF beta1 to the synovium is thus not suitable for treating intra-articular pathologies. PMID- 12723987 TI - Smoking-gender interaction and risk for rheumatoid arthritis. AB - The present case-control study was conducted to investigate the relationship between smoking and rheumatoid arthritis, and to investigate formally the interaction between sex, smoking, and risk for developing rheumatoid arthritis. The study was performed in the Central District of Finland. Cases were patients with rheumatoid arthritis and the control group was a random sample of the general population. Logistic regression models were used to evaluate the effect of smoking on risk for rheumatoid arthritis, after adjusting for the effects of age, education, body mass index, and indices of general health and pain. Overall, 1095 patients with rheumatoid arthritis and 1530 control individuals were included. Patients were older, less well educated, more disabled, and had poorer levels of general health as compared with control individuals (all P < 0.01). Preliminary analyses revealed the presence of substantial statistical interaction between smoking and sex (P < 0.001). In separate multivariable analyses, past history of smoking was associated with increased risk for rheumatoid arthritis overall in men (odds ratio 2.0, 95% confidence interval 1.2-3.2) but not in women. Among men, this effect was seen only for rheumatoid factor-positive rheumatoid arthritis. There were significant interactions between smoking and age among women but not among men. We conclude that sex is a biologic effect modifier in the association between smoking and rheumatoid arthritis. The role of menopause in the etiology of rheumatoid arthritis merits further research. PMID- 12723984 TI - Efficacy of treatment with glycosaminoglycans on experimental collagen-induced arthritis in rats. AB - To evaluate the antioxidant activity of the glycosaminoglycans hyaluronic acid (HYA) and chondroitin-4-sulphate (C4S), we used a rat model of collagen-induced arthritis (CIA). Arthritis was induced in Lewis rats by multiple intradermal injections of 250 microl of emulsion containing bovine type II collagen in complete Freund's adjuvant at the base of the tail and into three to five other sites on the back. Rats were challenged again with the same antigen preparation 7 days later. Disease developed about 11 days after the second immunization. The effects of treatment in the rats were monitored by biochemical parameters and by macroscopic and histological evaluations in blood, synovial tissue and articular cartilage. Arthritis produced the following symptoms: severe periarticular erythema, edema and inflammation in the hindpaws; membrane peroxidation in the cartilage of the joints; endogenous antioxidant wasting; high tumour necrosis factor-alpha (TNF-alpha) plasma levels; and synovial neutrophil accumulation. Treatment with HYA and C4S, starting at the onset of arthritis for 10 days, limited the erosive action of the disease in the articular joints of knee and paw, reduced lipid peroxidation, restored the endogenous antioxidants reduced glutathione (GSH) and superoxide dismutase, decreased plasma TNF-alpha levels, and limited synovial neutrophil infiltration. These data confirm that erosive destruction of the joint cartilage in CIA is due at least in part to free radicals released by activated neutrophils and produced by other biochemical pathways. The beneficial effects obtained with the treatment suggest that HYA and C4S could be considered natural endogenous macromolecules to limit erosive damage in CIA or as a useful tool with which to study the involvement of free radicals in rheumatoid arthritis. PMID- 12723988 TI - Osteoclast-independent bone resorption by fibroblast-like cells. AB - To date, mesenchymal cells have only been associated with bone resorption indirectly, and it has been hypothesized that the degradation of bone is associated exclusively with specific functions of osteoclasts. Here we show, in aseptic prosthesis loosening, that aggressive fibroblasts at the bone surface actively contribute to bone resorption and that this is independent of osteoclasts. In two separate models (a severe combined immunodeficient mouse coimplantation model and a dentin pit formation assay), these cells produce signs of bone resorption that are similar to those in early osteoclastic resorption. In an animal model of aseptic prosthesis loosening (i.e. intracranially self stimulated rats), it is shown that these fibroblasts acquire their ability to degrade bone early on in their differentiation. Upon stimulation, such fibroblasts readily release acidic components that lower the pH of their pericellular milieu. Through the use of specific inhibitors, pericellular acidification is shown to involve the action of vacuolar type ATPases. Although fibroblasts, as mesenchymal derived cells, are thought to be incapable of resorbing bone, the present study provides the first evidence to challenge this widely held belief. It is demonstrated that fibroblast-like cells, under pathological conditions, may not only enhance but also actively contribute to bone resorption. These cells should therefore be considered novel therapeutic targets in the treatment of bone destructive disorders. PMID- 12723986 TI - Functional analysis of an arthritogenic synovial fibroblast. AB - Increasing attention has been directed towards identifying non-T-cell mechanisms as potential therapeutic targets in rheumatoid arthritis. Synovial fibroblast (SF) activation, a hallmark of rheumatoid arthritis, results in inappropriate production of chemokines and matrix components, which in turn lead to bone and cartilage destruction. We have demonstrated that SFs have an autonomous pathogenic role in the development of the disease, by showing that they have the capacity to migrate throughout the body and cause pathology specifically to the joints. In order to decipher the pathogenic mechanisms that govern SF activation and pathogenic potential, we used the two most prominent methods of differential gene expression analysis, differential display and DNA microarrays, in a search for deregulated cellular pathways in the arthritogenic SF. Functional clustering of differentially expressed genes, validated by dedicated in vitro functional assays, implicated a number of cellular pathways in SF activation. Among them, diminished adhesion to the extracellular matrix was shown to correlate with increased proliferation and migration to this matrix. Our findings support an aggressive role for the SF in the development of the disease and reinforce the perspective of a transformed-like character of the SF. PMID- 12723989 TI - Autoantibodies to low-density-lipoprotein-receptor-related protein 2 (LRP2) in systemic autoimmune diseases. AB - We previously reported that autoantibodies (autoAbs) to the main epitope on CD69 reacted to its homologous amino acid sequence in low-density-lipoprotein-receptor related protein 2 (LPR2), a multiligand receptor for protein reabsorption. In this study, we have investigated the prevalence, autoepitope distribution, and clinical significance of the autoAbs to LRP2 in patients with systemic autoimmune diseases. Using six recombinant proteins (F2-F7) for LRP2 and one for CD69, we detected autoAbs to LRP2 in sera of patients with rheumatoid arthritis (RA), systemic lupus erythematosus, Behcet's disease, systemic sclerosis, and osteoarthritis and then mapped autoepitopes by Western blotting. The autoAbs to LRP2 were detected in 87% of the patients with rheumatoid arthritis, 40% of those with systemic lupus erythematosus, 35% of those with systemic sclerosis, 15% of those with osteoarthritis, and 3% of those with Behcet's disease. Multiple epitopes on LRP2 were recognized by most of the anti-LRP2+ serum samples. All of the tested anti-CD69 autoAb+ samples reacted to LRP2-F3 containing the homologous sequence to the main epitope of CD69; however, only 38% of the anti-LRP2-F3+ samples reacted to CD69. Clinically, the existence of the autoAbs to LRP2-F4, F5, and -F6 correlated with the presence of proteinuria in RA. This study revealed that LRP2 is a major autoantigen in RA. The autoAbs to LRP2 are probably produced by the antigen-driven mechanism and the autoimmunity to LRP2 may spread to include CD69. The anti-LRP2 autoAbs may play pathological roles by inhibiting the reabsorbing function of LRP2. PMID- 12723990 TI - The relation of menarcheal age to obesity in childhood and adulthood: the Bogalusa heart study. AB - BACKGROUND: Several studies have shown that girls who undergo menarche at a relatively young age tend to be more obese as adults. However, because childhood (pre-menarcheal) levels of weight and height are associated with an earlier menarche, the increased prevalence of adult obesity among early maturers may largely reflect the persistence of childhood obesity into adulthood. METHODS: We examined these interrelationships among 1179 girls (65% white, 35% black) who were examined as children (mean age, 9 y), adolescents, and adults (mean age, 26 y) in the Bogalusa Heart Study. RESULTS: Both white and black women who reported that they underwent menarche before age 12 y had, on average, higher adult levels of weight (+10 kg), body mass index (BMI, +4 kg/m2) and skinfold thicknesses (+6 mm) than did women who underwent menarche after age 13.5 y. However, relatively fat children tended to undergo menarche earlier than did thinner children, with each standard deviation increase in pre-menarcheal BMI increasing the odds of early menarche (<12 y) by approximately 2-fold. Stratified and regression analyses indicated that (1) adult obesity was more strongly associated with childhood obesity than with menarcheal age, and (2) about 60% to 75% of the apparent effect of menarcheal age was due to the influence of childhood obesity on both menarcheal age and adult obesity. CONCLUSIONS: Although additional longitudinal studies are needed, it is likely that the importance of early menarche in adult obesity has been overestimated. Most of apparent influence of menarcheal age on adult obesity is attributable to the association of childhood obesity with both menarcheal age and adult obesity. PMID- 12723991 TI - High prevalence of high risk human papillomavirus-capsid antibodies in human immunodeficiency virus-seropositive men: a serological study. AB - BACKGROUND: Serological study of human papillomavirus (HPV)-antibodies in order to estimate the HPV-prevalence as risk factor for the development of HPV associated malignancies in human immunodeficiency virus (HIV)-positive men. METHODS: Sera from 168 HIV-positive men and 330 HIV-negative individuals (including 198 controls) were tested using a direct HPV-ELISA specific to HPV-6, 11, -16, -18, -31 and bovine PV-1 L1-virus-like particles. Serological results were correlated with the presence of HPV-associated lesions, the history of other sexually transmitted diseases (STD) and HIV classification groups. RESULTS: In HIV-negative men low risk HPV-antibodies were prevailing and associated with condylomatous warts (25.4%). Strikingly, HIV-positive men were more likely to have antibodies to the high-risk HPV types -16, -18, -31, and low risk antibodies were not increased in a comparable range. Even those HIV-positive heterosexual individuals without any HPV-associated lesions exhibited preferentially antibody responses to the oncogenic HPV-types (cumulative 31.1%). The highest antibody detection rate (88,8%) was observed within the subgroup of nine HIV-positive homosexual men with anogenital warts. Three HIV-positive patients had HPV associated carcinomas, in all of them HPV-16 antibodies were detected. Drug use and mean CD4-cell counts on the day of serologic testing had no influence on HPV IgG antibody prevalence, as had prior antiretroviral therapy or clinical category of HIV-disease. CONCLUSION: High risk HPV-antibodies in HIV-infected and homosexual men suggest a continuous exposure to HPV-proteins throughout the course of their HIV infection, reflecting the known increased risk for anogenital malignancies in these populations. The extensive increase of high risk antibodies (compared to low risk antibodies) in HIV-positive patients cannot be explained by differences in exposure history alone, but suggests defects of the immunological control of oncogenic HPV-types. HPV-serology is economic and can detect past or present HPV-infection, independently of an anatomical region. Therefore HPV serology could help to better understand the natural history of anogenital HPV infection in HIV-positive men in the era of antiretroviral therapy. PMID- 12723993 TI - A case-control study of vanishing twin as a risk factor for cerebral palsy. AB - It has been hypothesized that cerebral palsy of unknown etiology is the result of the death of an unrecognized co-twin--a vanishing twin--in early gestation. We conducted a case-control study of vanishing twin as a risk factor for cerebral palsy of unknown etiology in women who had an obstetric ultrasound during pregnancy. Among mothers of cases, one of 86 had evidence of a vanishing twin on ultrasound, as compared to two of 381 control mothers (odds ratio [OR] 2.2, 95% confidence interval [CI] 0.2-24.8; p = 0.5). Bleeding in early pregnancy, which may indicate the loss of a co-twin, was reported by 14 case mothers and 46 control mothers (OR 1.6, 95% CI 0.8-3.0; p = 0.3). On the basis of results presented here, the vanishing twin syndrome is unlikely to account for a high proportion of cases of cerebral palsy, but there is insufficient statistical power to draw firm conclusions. PMID- 12723994 TI - Reliability and stability of mothers' reports about their pregnancies with twins. AB - The objective of this study was to determine if mothers' retrospective reports about events in their pregnancies with twins are reliable and stable. Six hundred and twenty-four mothers completed psychiatric interviews about their twins. These interviews also contained questions about the mothers' pregnancies, the perinatal period, and the child's early development. The mothers reported first on one twin and then on the other with interviews spaced from 3 days to 2 weeks apart. Thus mothers reported on the same pregnancy twice. Of these mothers, 47 were re interviewed 6 to 18 months later by raters blind to the results of the initial interview. The twin design allowed us to compare the short-term reliability of the 624 mothers' reports of the same pregnancy. The re-interview of the 47 mothers enabled us to compare the stability of reports over a longer time period. Agreement between the reports was measured with the kappa statistic. Kappas were good to excellent for the short-term reports of pregnancy for each twin for the 624 mothers. Kappas were equally high for the 47 mothers that were re-interviewed 6 to 18 months later. Mothers show good reliability and stability of reporting about events during pregnancy. PMID- 12723995 TI - Comparison of child-rearing problems between mothers with multiple children who conceived after infertility treatment and mothers with multiple children who conceived spontaneously. AB - The purpose of this study was to clarify the characteristic child-rearing problems for mothers of multiple children who conceived after infertility treatment as compared to mothers of multiple children who conceived spontaneously. The subjects were 990 mothers of multiple children: 359 who conceived after infertility treatment and 631 who conceived spontaneously. Mothers who conceived after infertility treatment were more delighted when informed of a multiple pregnancy than those who conceived spontaneously. In addition, with respect to anxiety during twin pregnancies, mothers of twins who conceived after infertility treatment showed lower rates of anxiety about nursing the infants and economic concerns than those of twins who conceived spontaneously. However, after delivery, mothers who conceived after infertility treatment showed a higher rate of depressive symptoms than those who conceived spontaneously. After adjusting for each associated factor using logistic regression, the risk of depressive symptoms in mothers who conceived after infertility treatment was significantly associated with disabled multiple children and the methods for alleviating stress. The odds ratio indicated that mothers with at least one disabled child were twice as likely to have depressive symptoms as mothers with no disabled children. Furthermore, the odds ratio indicated that mothers who used no methods for alleviating stress were twice as likely to have depressive symptoms than those who did. PMID- 12723992 TI - A novel Leishmania infantum nuclear phosphoprotein Lepp12 which stimulates IL1 beta synthesis in THP-1 transfectants. AB - BACKGROUND: We report cloning and characterization of a novel Leishmania infantum protein which we termed Lepp12, and we examine its possible implication in the interference with intramacrophage signaling pathways. RESULTS: The protein Lepp12 contains 87 amino acid sequence and exhibits 5 potential phosphorylation sites by protein kinase C (PKC). Recombinant GST-Lepp12 is phosphorylated in vitro by exogenous PKC and by PKC-like activities present in promastigote and in the myelomonocytic THP-1 cell line, indicating that at least one phosphorylation site is functional on the recombinant Lepp12. The natural Lepp12 protein is present in L. infantum promastigotes, as evidenced using specific anti-Lepp12 antibodies produced by immunopurification from acute phase VL patient sera. Interestingly, human patient sera are strongly reactive with GST-Lepp12, demonstrating immunogenic properties of Lepp12 in man, but no immune response to Lepp12 is detectable in experimentally infected animals. When isolated from promastigotes, Lepp12 migrates as two species of apparent MW of 18.3 kDa (major) and 14 kDa (minor), localizes in the nuclear fraction and appears constitutively phosphorylated. Natural Lepp12 is phosphorylable in vitro by both exogenous PKC and PKC-like activity present in THP-1 extracts. The intracellular Lepp12 transfected into THP-1 cells activates these cells to produce IL-1beta and induces an enhancing effect on PMA stimulated IL-1beta synthesis, as demonstrated using GST-Lepp12 transfectants. CONCLUSIONS: Together these results indicate that Lepp12 represents a substrate for PKC or other PKC-like activities present in the promastigote form and the host cell and therefore may interfere with signal transduction pathways involving PKC. PMID- 12723996 TI - Twins and non-twin siblings: different estimates of shared environmental influence in early childhood. AB - Twin studies typically indicate shared environmental influence for cognitive abilities, especially in early childhood. However, across studies, DZ twin correlations tend to be greater than non-twin sibling correlations, suggesting that twin estimates of shared environment are to some extent specific to twins. We tested this hypothesis in a sample of more than 1800 MZ and 1800 same-sex DZ pairs from the Twins Early Development Study (TEDS), a study of twins born in England and Wales in 1994 and 1995. For this analysis, we obtained comparable data from more than 130 same-sex younger siblings of the twins. Twins and their younger siblings were assessed for language, cognitive abilities and behavior problems by their parents at 2 and 3 years of age. For language and cognitive measures at both 2 and 3 years, but not for behavior problems, estimates of shared environment were more than twice as large for twins as compared to non twin siblings. We conclude that about half of twin study estimates of shared environment for cognitive abilities in early childhood are specific to twins. Although many possibilities exist for explaining the special shared environment effect for twins, we suggest that cognitive-relevant experiences that are not shared by siblings are shared by twins because they are exactly the same age. PMID- 12723997 TI - Peer reports of adaptive behavior in twins and singletons: is twinship a risk or an advantage? AB - We compared twins to their gender-matched singleton classmates in peer-assessed behavioral adjustment. Our samples include 1874 11- to 12-year-old Finnish twins (687 monozygotic, MZ; 610 same-sex dizygotic, SSDZ; 577 opposite-sex dizygotic, OSDZ) and their 23,200 non-twin classmates. Data were collected using a 30-item Multidimensional Peer Nomination Inventory containing three factors and their subscales. We found twin-singleton differences: classmates rated twin girls and boys higher than gender-matched singletons in Adaptive Behaviors (constructive, compliant, and socially active behavior), and those effects were particularly evident among OSDZ twins for assessments of social interaction, popularity, and leadership. We found no evidence that individual twins differ from singletons in Externalizing (hyperactivity-impulsivity, inattention, aggression) or Internalizing Problem Behaviors (depressive symptoms, social anxiety). Nor did we find systematic differences between MZ and SSDZ twins. Among both twins and singletons, boys exceeded girls in Externalizing, and girls exceeded boys in Internalizing Problem Behaviors. Results suggest that a twinship forms a positive developmental environment for socioemotional behavior, particularly among OSDZ twins. PMID- 12723998 TI - Special twin environments, genetic influences and their effects on the handedness of twins and their siblings. AB - It has been suggested that twinning may influence handedness through the effects of birth order, intra-uterine crowding and mirror imaging. The influence of these effects on handedness (for writing and throwing) was examined in 3657 Monozygotic (MZ) and 3762 Dizygotic (DZ) twin pairs (born 1893-1992). Maximum likelihood analyses revealed no effects of birth order on the incidence of left-handedness. Twins were no more likely to be left-handed than their singleton siblings (n = 1757), and there were no differences between the DZ co-twin and sibling-twin covariances, suggesting that neither intra-uterine crowding nor the experience of being a twin affects handedness. There was no evidence of mirror imaging; the co twin correlations of monochorionic and dichorionic MZ twins did not differ. Univariate genetic analyses revealed common environmental factors to be the most parsimonious explanation of familial aggregation for the writing-hand measure, while additive genetic influences provided a better interpretation of the throwing hand data. PMID- 12723999 TI - Genetic correlations between brain volumes and the WAIS-III dimensions of verbal comprehension, working memory, perceptual organization, and processing speed. AB - We recently showed that the correlation of gray and white matter volume with full scale IQ and the Working Memory dimension are completely mediated by common genetic factors (Posthuma et al., 2002). Here we examine whether the other WAIS III dimensions (Verbal Comprehension, Perceptual Organization, Processing Speed) are also related to gray and white matter volume, and whether any of the dimensions are related to cerebellar volume. Two overlapping samples provided 135 subjects from 60 extended twin families for whom both MRI scans and WAIS III data were available. All three brain volumes are related to Working Memory capacity (r = 0.27). This phenotypic correlation is completely due to a common underlying genetic factor. Processing Speed was genetically related to white matter volume (r(g) = 0.39). Perceptual Organization was both genetically (r(g) = 0.39) and environmentally (r(e) = -0.71) related to cerebellar volume. Verbal Comprehension was not related to any of the three brain volumes. It is concluded that brain volumes are genetically related to intelligence which suggests that genes that influence brain volume may also be important for intelligence. It is also noted however, that the direction of causation (i.e., do genes influence brain volume which in turn influences intelligence, or alternatively, do genes influence intelligence which in turn influences brain volume), or the presence or absence of pleiotropy has not been resolved yet. PMID- 12724000 TI - A twin study of genetic influences on epilepsy outcome. AB - The identification of genetic factors that confer susceptibility to the epilepsies has to date been the focus of genetic efforts in this field. Few studies have assessed the genetic contribution to disease course in epilepsy, yet an understanding of the genetic influences on epilepsy outcome is key to developing new therapeutic strategies. The aim of this study was to assess the genetic contributions to epilepsy outcome in twin pairs concordant for epilepsy. We studied 37 epilepsy concordant twin pairs (27 monozygotic, 10 dizygotic) in whom there were no recognized environmental contributions (e.g., acquired brain injury) to epilepsy, and in whom the most likely cause for epilepsy was a shared genetic susceptibility. Clinical outcome was determined using the binary measure of Seizure Status (seizure remission or recurrence) and on a six-category ordinal Outcome Scale. Epilepsy outcome was independent of age of seizure onset, age at assessment and major epilepsy syndrome diagnosis. The proportion of twin pairs concordant for Seizure Status was 0.81 (22/27) for monozygous and 1.0 (10/10) for dizygous pairs, p = 0.3. Within-pair correlation in outcome (Outcome Scale) was 0.60 (95% CI: 0.32, 0.78) in monozygous and 0.78 (0.48, 0.92) in dizygous pairs. These data provide no evidence for genetic influences on epilepsy outcome independent of those that contribute to disease susceptibility. The observed high correlations for outcome suggest that, for epilepsy, susceptibility genes also have a major influence on outcome. PMID- 12724001 TI - Primary osteoarthritis of the hip in monozygotic and dizygotic male twins. AB - Population and total hip replacement surveys show that primary osteoarthritis of the hip is uncommon in African Americans and rare in Asians, suggesting a genetic basis for this disease. We studied genetic influences on primary osteoarthritis of the hip by estimating monozygotic (MZ) and dizygotic (DZ) twin correlations using a two-stage data collection. A total of 6419 male veteran twins of the NAS NRC Twin Registry, born between 1917 and 1927, were contacted by telephone (first stage). Telephone interview determined that 2% reported a total hip replacement for arthritis rather than fracture. X-rays of twin pairs in which one twin had undergone total hip replacement were sought and reviewed (second stage), and concordance for primary arthritis was determined based on x-ray diagnosis. Heritability of primary osteoarthritis, Kellgren & Lawrence Grade II and higher, was estimated using a covariance structure analysis for 2-stage data. The best fitting model included only components for additive genetics and for unique environment. Additive genetics accounted for 53% (95% confidence interval 30-72%) in the liability for self reported hip replacement surgery and unique environment for the remaining 47% (95% confidence interval 28-70%). Additive genetics accounted for 61% (95% confidence interval 18-86%) of the variance in liability for x-ray determined primary osteoarthritis with unique environment accounting for the remaining 39%. These data establish a genetic influence on primary osteoarthritis of the hip in male twins and suggest that further work is indicated to isolate the genes responsible for this disease. PMID- 12724002 TI - The relation between insulin resistance and hemostasis: pleiotropic genes and common environment. AB - Risk factors for coronary heart disease (CHD), including prethrombotic changes in hemostasis, cluster with the insulin resistance (IR) syndrome. The aim of the present study was to investigate to what extent the relation between IR and hemostatic risk factors is due to shared genes or environmental factors. Multivariate genetic analysis was performed using a total of 314 (107 monozygotic and 207 dizygotic) twin pairs on IR assessed by HOMA, fibrinogen, plasminogen activator inhibitor (PAI-1), tissue plasminogen activator (tPA), factor VIII (FVIII), von Willebrand factor (vWF) and factor XIII B-subunit. The relationship between IR and the 6 hemostatic factors could best be explained by an independent pathway model consisting of 2 common genetic factors, one of which influenced IR and all hemostatic factors, and 3 common environmental factors, each representing the shared variance between IR and different aspects of the hemostatic system. Genetic correlations between IR and hemostatic proteins were larger than their environmental counterparts. Since IR and prethrombotic changes are features of both diabetes and CHD, the finding of one set of pleiotropic genes warrants the identification of these common pathways which may provide new avenues for treatment and prevention of both diabetes and CHD. PMID- 12724004 TI - Immune reconstitution syndrome in AIDS. Interview by Vicki Glaser. PMID- 12724003 TI - No linkage to obesity in candidate regions of chromosome 2 and 10 in a selected sample of Swedish twins. AB - The aim of the current study was to investigate the importance of genetic and environmental effects in the variation of body mass index, and to investigate linkage for obesity to previously reported candidate regions on chromosome 2 and 10. A sample of 1422 twin pairs from the population based Swedish Twin Registry was used in order to estimate the genetic and environmental effects in the variation of body mass index by means of structural equation modeling. A selection of those, 51 concordant and 155 discordant for obesity, was used for the linkage analysis by implementing the "combined" Haseman-Elston approach. Heritability of body mass index ranged from 59-70%, implying that genetic effects were of importance for the variation of obesity, and there were significant sex and age differences. Linkage could not be verified in candidate regions of chromosomes 2 and 10, indicating that these genetic variants have a significant effect in extreme obese populations rather than in moderately obese Caucasians. However, the results were sensitive to issues related to power, minor effects of the genes, ethnic differences and the complex mechanism underlying obesity. PMID- 12724005 TI - Indinavir-induced thrombocytopenia. PMID- 12724006 TI - Prospective comparison of first-line nelfinavir therapy versus nelfinavir introduction in rescue antiretroviral regimens. AB - In order to establish the role of the protease inhibitor nelfinavir in current clinical practice, a prospective 18-month open-label comparison of efficacy and tolerability of nelfinavir was performed among HIV-infected patients who either incorporated nelfinavir in their first-line highly active antiretroviral therapy (HAART) regimen (group A, 57 patients), or who added nelfinavir to a rescue antiretroviral regimen (following at least two attempts with protease inhibitor based HAART) (group B, 67 patients). All evaluable data were analyzed according to the prior and concurrent antiretroviral therapy, including genotypic resistance assays for patients undergoing salvage therapy. A significantly better virologic outcome (as expressed by a > 2 log(10) drop of plasma viremia versus baseline or attainment of undetectable levels), was shown among patients belonging to group A versus group B, where a number of genotypic mutations possibly elicited by previous anti-HIV treatment strongly impaired a potent and sustained nelfinavir activity. On the whole, the immunologic response (as expressed by the mean CD4(+) lymphocyte count versus baseline), substantially paralleled the virologic one in all analyzed subgroups, but a tendency toward a maintained immunologic competence was also observed in the majority of patients experiencing virologic failure. Nelfinavir introduction was sufficiently safe, because a limited percentage of patients suffered from mild-to-moderate, novel, or continuing adverse events, which proved significantly more frequent in the salvage group but did not affect adherence to HAART. PMID- 12724007 TI - The impact of apathy and depression on quality of life in patients infected with HIV. AB - Apathy refers to decreased self-initiation and goal-directed behavior. Apathy is a relatively common neuropsychiatric symptom associated with HIV, yet the impact of apathy on health-related quality of life (QOL) has not been investigated. We examined the relationship between apathy, depression, and QOL among individuals infected with HIV. Apathy was quantified using the Marin Apathy scale and QOL was measured with the Medical Outcomes Study Short-Form 36 (SF-36). Results of the study revealed that both apathy and depression were more common among patients with HIV than healthy control subjects. Twenty-six percent of the patients with HIV reported clinically significant apathy while 80% of the patients reported clinically significant depression. Apathy did not relate to ratings of overall QOL, but it was modestly associated with ratings of mental health and role disruption secondary to mental health. By contrast, ratings of depression were strongly related to overall QOL and most indices of SF-36. Regression equations revealed that depression and apathy independently contributed to mental health and role disruption secondary to mental health. Importantly, ratings of depression accounted for the majority of variance for ratings of QOL. The findings indicate that while apathy is more common among individuals with HIV compared to healthy control subjects, the impact of apathy on QOL is less significant than depression. Clinicians should continue to focus on depression as an important neuropsychiatric symptom associated with HIV. PMID- 12724008 TI - Survey of physician attitudes toward HIV testing in pregnant women in Ohio. AB - HIV infection among women of childbearing age is still increasing in the United States. In most states, HIV testing of women or neonates during pregnancy is not mandatory. The current study assessed HIV prenatal testing practices among obstetrician-gynecologists and primary care physicians listed in a regional physician referral data base in a predominantly rural region. Between December 2000 and March 2001 a 20-question survey was sent by mail to regional physicians in obstetrics/gynecology and primary care regarding physician practice demographics and prenatal HIV testing practices. Of 1116 surveys sent, 431 were returned (38.6% response). Only 42% of physicians offered universal HIV prenatal testing. Factors associated with universal testing (p < 0.5) included obstetrics/gynecology as the practice specialty (90%) physicians' age younger than 50 years, and a practice with predominantly Medicaid or African American patients. Further educational and public health initiatives may be needed to increase nonselective, universal HIV testing in pregnant women. PMID- 12724009 TI - Imaging HIV/AIDS. Body cavity-based lymphoma. AB - Non-Hodgkin's lymphoma (NHL) is an important and common AIDS-related neoplasm. AIDS-related NHL can be defined by its anatomic distribution as systemic (nodal and extranodal masses), primary central nervous system, and occasionally body cavity-based lymphomas (BCBL). Radiologic imaging plays an important role in differentiating the varied appearances of AIDS-related NHL, particularly regarding BCBL: patients' images demonstrate only fluid in the pleural, pericardial, and/or peritoneal spaces. Imaging of a case of BCBL is presented, including simultaneous pericardial, pleural, and peritoneal effusions. PMID- 12724010 TI - Daily pill burden and adherence. PMID- 12724011 TI - Black media sponsor prevention program. PMID- 12724012 TI - Proliferative activity in primary ovarian carcinoid tumors. AB - The proliferative potential of six primary ovarian carcinoids with different clinical outcome and histogenetic origin was examined immunohistochemically. The results showed that two cases with extremely high level of proliferative activity were associated with metastatic spread. In the remaining tumors, the examined factor was found to be at low level comparable with excellent prognosis of typical carcinoids in other locations. The preliminary results showed a possibility of a prognosis prediction according to typing of the ovarian carcinoids into two categories, i.e., tumors of low and intermediate malignancy. Topoisomerase II-alpha and Ki-67 are suitable markers giving valuable information about this phenomenon. PMID- 12724013 TI - Leptin, neuropeptide Y, beta-endorphin, gonadotropin, and estradiol levels in girls before menarche. AB - More than two decades of clinical studies have provided us with the opportunity to develop and implement criteria that distinguish three phases during pre menarche, paralleling rising levels of estrogens, namely 'pre-estrogenization', 'onset of estrogenization' and 'full estrogenization'. The aim of this study was to examine the relationships between somatic features and levels of leptin, neuropeptide Y (NPY), beta-endorphin, gonadotropin and estradiol in pubertal girls before menarche. Weight, height, body mass index (BMI), tertiary sex features, estrogen-related changes in hymen, fat and lean body mass were studied on a quarterly basis in 45 girls. At the same time, ovarian and uterine dimensions were established sonographically and serum was obtained for the determination of leptin, NPY, beta-endorphin, gonadotropin, and estradiol levels. Onset of estrogenization in girls was marked by weight loss, followed by an increase in total, fat and lean body mass, and pubertal acceleration of growth. At the time of full estrogenization, lower NPY and beta-endorphin levels were observed, accompanied by an increase in gonadotropin secretion. Changes in leptin levels are consistent with a role of this hormone in metabolic signaling. PMID- 12724014 TI - Use of complementary therapies by women attending a specialist premenstrual syndrome clinic. AB - In this study, we investigate the use of complementary therapies by women attending a specialist premenstrual syndrome (PMS) clinic in the UK. Data was collected via an anonymous questionnaire survey of 100 women attending the clinic. Results showed 91% of women had used at least one form of complementary therapy for the management of their premenstrual symptomatology although only 35% were current users. Over half (53%) felt that these therapies had been of some benefit. Prescribed medication for PMS was being used by 71% of women at the time of the questionnaire and 83% of these women were satisfied with the perceived success of conventional therapy. In conclusion, the vast majority of women attending a specialist PMS clinic in the UK have used complementary therapies to treat this chronic debilitating condition but few continue use long-term. Treatment may be instigated by the woman with advice from her informal support network and/or her physicians. However as use is so prevalent, but with few randomized controlled trials conducted to show their benefits or risks, it is important to improve awareness of these therapies, both in qualitative and quantitative terms. Satisfaction with prescribed medications did not appear to be influenced by complementary therapy use in this group of women. PMID- 12724015 TI - Evaluation of bone turnover in postmenopausal patients with type 2 diabetes mellitus using biochemical markers and bone mineral density measurements. AB - Although osteoporosis is reported as a potential complication of type 1 diabetes mellitus (DM), the effects of type 2 DM on bone mass are conflicting. Most of the studies conducted in recent years reveal that bone mineral density (BMD) values of type 2 DM patients are not decreased and even increased when compared with healthy control groups. In this study we evaluated bone turnover in 57 postmenopausal type 2 DM patients utilizing biochemical markers for bone formation and resorption, and BMD measurements. We found that BMD values in diabetic patients (0.91 +/- 0.11 g/cm(2) for lumbar region, 0.89 +/- 0.14 g/cm(2) for hip region) were higher than healthy postmenopausal control group (0.81 +/- 0.12 g/cm(2) for lumbar region, 0.76 +/- 0.10 g/cm(2) for hip region). Serum alkaline phosphatase values were similar to the control group, whereas serum osteocalcin and N-telopeptide/creatinine (NTx/Cr) values were significantly lower than the control group (osteocalcin: 8.82 +/- 4.03 ng/ml, NTx/Cr: 122.70 +/- 81.76 nMBCE/mMCr) in diabetic patients (osteocalcin: 4.44 +/- 3.53 ng/ml, NTx/Cr: 42.24 +/- 29.97 nMBCE/mMCr). Also a significant correlation was observed between body mass index and BMD values. Our findings suggested that the bone turnover rate is remarkably lower in type 2 DM patients compared to healthy postmenopausal patients. PMID- 12724016 TI - Inverse correlation between serum leptin concentration and vertebral bone density in postmenopausal women. AB - Although the protective effect of obesity on bone mass has been ascribed to high body fat content, this is still controversial. The present study of 215 postmenopausal Japanese women investigated whether circulating leptin concentration was correlated with per cent fat mass or age-adjusted bone mineral density (Z-score). In this study population, the mean circulating leptin concentration was 10.15 +/- 5.42 (range 1.7-29.6) ng/ml. Significant correlations were found between circulating leptin concentrations and per cent fat mass (r = 0.514, p < 0.0001) and Z-score (r = -0.516, p < 0.0001), confirming the existence of an inverse relationship between leptin concentration and postmenopausal bone density. By contrast, no significant correlation was found between per cent abdominal fat mass and vertebral bone mineral density (r = -0.071). Serum leptin concentration reflects fat mass and postmenopausal bone mass in human subjects. Increased serum leptin levels might cause bone loss in postmenopausal Japanese women, and our results do not support the hypothesis that leptin induces or mediates the bone-protective effects of obesity. PMID- 12724017 TI - Biosynthesis of MUC1 mucin in human endometrial adenocarcinoma is modulated by estradiol and tamoxifen. AB - Polymorphic epithelial mucin MUC1 is expressed by most epithelial cancers, although free natural MUC1 antibodies are present in the circulation of healthy subjects as well as in that of cancer patients. The role of MUC1 mucin molecules in cancer cells of endometrium is not precisely known. The results reported here demonstrate that MUC1 biosynthesis in human endometrial adenocarcinoma cells (Ishikawa line) is stimulated by estradiol hormone and inhibited by tamoxifen, which was measured by [(14)C]threonine or [(3)H]glucosamine incorporation into MUC1 protein. Tamoxifen applied in combination with estradiol also inhibited this process, but pre-incubation of cells with estradiol resulted in a decrease in the inhibitory effect of tamoxifen. Electroblotting and reactions with antibodies against MUC1 core protein epitopes confirmed the presence of MUC1 in cell lysates and culture media of Ishikawa cells. Reactions with lectins showed the presence of oligosaccharide structures demonstrating antigen-T activity and the presence of sialic acid residues. The results confirm that there is downregulation of MUC1 expression in cancer culture cells treated with selective estrogen receptor modulators, which may be essential for reducing the migration of cancer cells and the metastatic properties of tumor cells. PMID- 12724018 TI - Effects of genistein on the endometrium: ultrasonographic evaluation. AB - The aim of our study was to evaluate the effects of isoflavones on climacteric related symptoms and on the endometrium in postmenopausal women, in a prospective, open, randomized, clinical trial performed at the Menopause Clinic of our Department. Seventy postmenopausal women were randomly assigned to two treatment groups receiving 12 cycles of treatment with genistein (group A) or calcium (group B). In all patients ultrasonographic endometrial thickness and Kupperman Index (KI) were evaluated at baseline and after 6 and 12 cycles of treatment. At baseline no significant difference was detected in endometrial thickness and in KI between groups A and B. After 6 and 12 cycles of treatment, no significant difference was observed in endometrial thickness between or within groups. Endometrial thickness was lower than 5 mm in all cases before and during treatment except in two cases in group B and in one case in group A after 12 months. At 6 and 12 months, the KI was significantly (p < 0.05) lower in group A in comparison with baseline values and group B. We conclude that genistein administration reduces climacteric symptoms in postmenopausal women and does not increase endometrial thickness. PMID- 12724019 TI - Effects of metformin therapy on hyperandrogenism in women with polycystic ovarian syndrome. AB - Polycystic ovarian syndrome (PCOS) is one of the most common endocrine diseases in women. This syndrome is characterized by hyperandrogenism, chronic anovulation, infertility and obesity. The association between PCOS-related hyperandrogenemia and insulin resistance is well documented in the literature. Insulin resistance and the resulting raised plasma levels of insulin are reported to be responsible for the high androgen concentration observed in patients with PCOS. In this prospective study, blood samples for levels of testosterone (T), dehydroepiandrosterone sulfate (DHEAS), luteinizing hormone (LH), follicle stimulating hormone (FSH), LH/FSH, prolactin and fasting blood sugar (FBS) before starting metformin administration were obtained randomly from 40 women who were apparently obese, had PCOS and had been referred to a university hospital. Metformin was then given at a dose of 500 mg three times a day for 8 weeks, after which time the pretreatment study was repeated. Clinical symptoms of PCOS, including acne and hirsutism score and body mass index (BMI), were assessed before and after the treatment cycle. Metformin therapy resulted in a significant decrease in total testosterone levels and FBS. There was also a significant decline in BMI, length of the menstrual cycle, acne and hirsutism score. There were no significant changes in the levels of DHEAS, prolactin, FSH or LH, or in LH/FSH. The effect of metformin on subjects with elevated DHEAS levels was different to that on individuals with normal DHEAS levels. In the latter group there were only significant improvements in the length of the menstrual cycle, BMI and testosterone and DHEAS levels. It is concluded that metformin therapy in subjects with PCOS results in a decrease in fasting blood sugar and testosterone levels, and leads to a significant improvement in the clinical manifestation of hyperandrogenism. These responses also related to the level of adrenal function. PMID- 12724021 TI - Effect of different hormonal replacement therapies on circulating allopregnanolone and dehydroepiandrosterone levels in postmenopausal women. AB - The effects of hormone replacement therapy (HRT) on the central nervous system in postmenopausal women might be mediated by changes in neurosteroid synthesis and/or release. The aim of this study was to evaluate the impact of HRT on the levels of allopregnanolone, a sedative anxiolytic GABA(A) agonist steroid, and dehydroepiandrosterone (DHEA), a GABA(A) antagonist steroid. We evaluated allopregnanolone and DHEA circulating levels after 1, 3, 6, 9 and 12 months of HRT with ten different estrogen or estrogen-progestin molecules, regimens and routes of administration in 186 postmenopausal women. Cortisol, luteinizing hormone, follicle stimulating hormone, estradiol and progesterone levels were also evaluated. Allopregnanolone levels significantly increased during follow-up with all HRT preparations. The addition of progestin molecules (except for 19-nor derivatives) to transdermal estradiol administration alone determined a higher increase in allopregnanolone levels. Transdermal HRT showed a significantly higher percentage change in allopregnanolone levels compared with oral HRT. DHEA levels showed a progressive decline starting from the 3-month follow-up, without significant differences between the transdermal and oral groups, as well as among the ten groups, independently of the presence and type of progestin molecule used. In conclusion, HRT strongly modifies circulating neurosteroid levels in postmenopausal women. PMID- 12724022 TI - Thyroid function and postmenopause. AB - There is an increasing prevalence of high levels of thyroid stimulating hormone (TSH) with age - particularly in postmenopausal women - which are higher than in men. The incidence of thyroid disease in a population of postmenopausal women is as follows: clinical thyroid disease, about 2.4%; subclinical thyroid disease, about 23.2%. Among the group with subclinical thyroid disease, 73.8% are hypothyroid and 26.2% are hyperthyroid. The rate of thyroid cancer increases with age. The symptoms of thyroid disease can be similar to postmenopausal complaints and are clinically difficult to differentiate. There can also be an absence of clinical symptoms. It is of importance that even mild thyroid failure can have a number of clinical effects such as depression, memory loss, cognitive impairment and a variety of neuromuscular complaints. Myocardial function has been found to be subtly impaired. There is also an increased cardiovascular risk, caused by increased serum total cholesterol and low-density lipoprotein cholesterol as well as reduced levels of high-density lipoprotein. These adverse effects can be improved or corrected by L-thyroxine replacement therapy. Such treatment has been found to be cost-effective. With time, overt hypothyroidism can develop. Therefore, routine screening of thyroid function in the climacteric period to determine subclinical thyroid disease is recommended. Hormone replacement therapy (HRT) in women with hypothyroidism treated with thyroxine causes changes in free thyroxine and TSH. Increased binding of thyroxine to elevated thyroxine-binding globulin causes an elevation of TSH by feedback. Since adaptation is insufficient, there is an increased need for thyroxine in these women taking HRT. TSH levels should be controlled at 12 weeks after the beginning of therapy. At higher age the need for iodine and thyroxine is decreased. Therefore, therapy has to be controlled. For bone metabolism thyroid hormones play a dominant role. While there are only marginal differences between hypothyroid patients and euthyroid controls, there are large differences for hyperthyroid patients. Previous thyrotoxicosis and subsequent long-lasting L-thyroxine treatment are together associated with reduction in femoral and vertebral bone density in postmenopausal women. In these cases HRT is important for the control of bone loss. PMID- 12724020 TI - The benefits of finasteride for hirsute women with polycystic ovary syndrome or idiopathic hirsutism. AB - Finasteride has been used frequently in the treatment of prostate hyperplasia, but this drug inhibits 5alpha-reductase and for this reason could be useful for the treatment of hirsutism. The aim of this study was to evaluate the clinical and hormonal effects of finasteride on hirsute women with idiopathic hirsutism or polycystic ovary syndrome. Twenty-four women were randomly divided into two groups: those given placebo and those given finasteride 5 mg/day. The treatment period was 6 months. All patients were evaluated before the beginning of treatment (baseline) and after 3 and 6 months of treatment using clinical examination through Ferriman-Gallwey score, blood pressure, cardiac frequency and body mass index. Also, we collected blood for hormonal determination of levels of prolactin, 17alpha-hydroxyprogesterone, follicle stimulating hormone, luteinizing hormone, total and free testosterone, dehydroepiandrosterone sulfate, androstenedione and dihydrotestosterone. Furthermore, all patients were asked about their concerns and satisfaction with the treatment. The results showed that the Ferriman-Gallwey score in the 6th month of finasteride treatment was significantly lower than at baseline and the 3rd month of this drug treatment. The dihydrotestosterone level in the finasteride group was also significantly reduced compared to that in the placebo group. The other hormones did not show any statistical difference during the study. All the patients treated with finasteride perceived a reduction in hirsutism after 6 months. In conclusion, our data suggest that finasteride may be effective for the treatment of the hirsute woman with idiopathic hirsutism or polycystic ovary syndrome. PMID- 12724023 TI - The Regius Chair of Military Surgery in the University of Edinburgh, 1806-55. PMID- 12724032 TI - Personalised prescribing for asthma--is pharmacogenetics the answer? AB - An individual's response to anti-asthma medication is likely to arise from a complex interaction between social, environmental and inherited factors. Studies indicate that genetic factors may account for 60-80% of the heterogeneity in treatment responsiveness in asthmatics. Identifying the genetic variants responsible may potentially lead to the development of novel treatments, improved effectiveness in the use of existing treatments and better prediction of efficacy in phase II and III trials. This article will briefly outline the current methods of identifying relevant treatment-responsive genes and their genetic variants. The pharmacogenetics of the main asthma treatment groups will then be reviewed in detail. Finally, the impact of pharmacogenetics on the pharmaceutical industry, and clinical practice in the future will be discussed. PMID- 12724033 TI - Antimicrobial activity of beta-lactam antibiotics against clinical pathogens after molecular inclusion in several cyclodextrins. A novel approach to bacterial resistance. AB - Recognition and uptake by specific cellular receptors and transport systems for cyclodextrins have been demonstrated. Based on this concept, natural and synthetically modified cyclodextrins were used as drug carriers. Several beta lactam antibiotics were selected and their inclusion complexes with different cyclodextrins were prepared (molar ratio ranging from 1:1 to 1:3). The complex formation, in aqueous solution, was monitored and optimum complexation conditions were selected. The inclusion of the active molecules in the cyclodextrin cavity was confirmed by (1)H NMR spectroscopy. Specific HPLC methods for the quantitation of antibiotics in the presence of cyclodextrins were developed and their chemical stability under complexation conditions was confirmed. Antimicrobial activity of drug-cyclodextrin complexes, in terms of minimum inhibitory concentration (MIC), were compared with the corresponding values of uncomplexed free molecules. A wide range of clinical pathogens and known beta lactamase-producing strains were tested. The activity of the cyclodextrin included antibiotics was increased, particularly against Gram-negative clinical strains. The nature and degree of substitution on cyclodextrin macromolecules may be the predominant factor in the observed improvement in antimicrobial activity. We believe that the proposed methodology is a novel approach to the microbial resistance problem and will trigger research towards the development of new cyclodextrin derivatives bearing the ability to increase the uptake of included antimicrobial molecules through intensification of the corresponding molecular recognition phenomena. PMID- 12724034 TI - The effect of protective agents on the stability of plasmid DNA by the process of spray-drying. AB - The effect of several protective agents was assessed on the stability of spray dried plasmid DNA. The spray-drying process had adverse effects on the tertiary structure of plasmid DNA with the protective agents of sucrose, glycine and agarose. With the protection of these noncondensing agents, a band corresponding to the linear form of plasmid DNA was observed in the gel electrophoresis between the supercoiled circular (SC) form and the open circular (OC) form. On the contrary, spray-dried plasmid DNA maintained some degree of structural integrity under the protection of condensing agents. For the protection by neutral condensing polymers, such as polyethylene glycol 1000 and 4000, no linear form between the SC form and the OC form of plasmid DNA was revealed in the gel electrophoresis. Also, excess cationic condensing polymer, polyethyleneimine, had the ability to provide the plasmid DNA with protection from degradation as indicated by the preservation in SC and OC forms of plasmid DNA on the agarose gel electrophoresis. Moreover, DNA topology was unchanged after six-month storage at 4 degrees C by the protection of these neutral and cationic condensing agents. Accordingly, DNA condensation induced by condensing agents may provide a way to minimize damage to plasmid DNA by the process of spray drying. PMID- 12724035 TI - Resveratrol transport and metabolism by human intestinal Caco-2 cells. AB - Resveratrol is a dietary constituent suggested to have protective effects against cancer as well as cardiovascular disease. The purpose of the study was to learn whether this agent could be absorbed in man and enter the systemic circulation. This was examined by measuring transport and metabolism of resveratrol (5-40 microM) by the human intestinal epithelial cell line Caco-2 cultured in Transwells. Transport across the Caco-2 monolayer occurred in a direction independent manner with P(app) values of approximately 7 x 10(-6) cm s(-1), much higher than for the paracellular transport marker mannitol (approximately 0.4 x 10(-6) cm s(-1)), suggesting efficient absorption in-vivo. At the highest resveratrol concentration, the absorption increased, possibly due to saturation of metabolism. In sharp contrast to previous findings in the rat, the metabolism of resveratrol in Caco-2 cells involved mainly sulfation and, to a minor extent, glucuronidation. At low resveratrol concentrations, most of the sulfate conjugate was exported to the apical side, presumably by MRP2, which is well expressed in these cells. At high concentrations, there was a shift towards the basolateral side, possibly involving MRP3, which was recently shown also to be expressed in Caco-2 cells. These results indicate that absorption of resveratrol in-vivo may be high but with limited bioavailability due to efficient sulfate conjugation. Extensive accumulation of resveratrol in the Caco-2 cells, demonstrated in additional experiments, suggests enterocytes as a major target site for this cancer preventive agent. PMID- 12724036 TI - Influence of co-administered antibiotics on the pharmacokinetic fate in rats of paeoniflorin and its active metabolite paeonimetabolin-I from Shaoyao-Gancao tang. AB - The effects of orally co-administered antibiotics on the pharmacokinetics of paeoniflorin (PF) and paeonimetabolin-I (PM-I), a bioactive metabolite derived from PF by intestinal bacteria, from the traditional Chinese formulation, Shaoyao Gancao-tang (SGT), were investigated in rats to clarify the effect of administering SGT together with some synthetic drugs. Co-administration of the antibiotics amoxicillin and metronidazole (AMPC-MET) significantly increased the area under the plasma concentration versus time curve (AUC) of PF, whereas it markedly decreased that of PM-I, to 2.6% of the normal AUC by administration of a single dose, and less than 1% by a 3-day pretreatment. Similar effects were observed using the combination of ofloxacin with SGT. The PF-metabolizing activity of intestinal bacteria was reduced to 16% and 33% of normal levels by treatment with AMPC-MET and ofloxacin, respectively, which caused alterations of that degree in the extent of absorption of PF and PM-I, but did not affect their rate of absorption or elimination. The present study suggests that it may not be appropriate to use SGT simultaneously with antibiotics such as AMPC-MET or ofloxacin, and also reveals the important role of intestinal bacteria in the pharmacokinetics of the active components of this traditional Chinese formulation. PMID- 12724037 TI - Investigation on the antiplatelet activity of pyrrolo[3,2-c]pyridine-containing compounds. AB - A series of 4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridines (THPPs), mostly C(2) substituted derivatives, and some 2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indoles (THPIs) were synthesized and tested in-vitro for their ability to inhibit aggregation of human platelet-rich plasma (PRP) induced by adenosine 5' diphosphate (ADP) and adrenaline (epinephrine). 5-Benzyl THPP (3), 2 (benzylamino)methyl THPP (5f) and 2-ethyl THPI (6) moderately and dose dependently inhibited platelet aggregation induced by adrenaline and, to a lesser extent, by ADP. These compounds inhibited the second phase of the PRP aggregation triggered by adrenaline, which largely depends upon thromboxane A(2) production and ADP release. In the adrenaline stimulated aggregation, the THPI derivative 6 was found to be nearly equipotent with aspirin, their IC50 values (concentration effecting 50% inhibition of aggregation) being 90 and 60 microM, respectively. A relation between activity and calculated octanol-water partition coefficient suggested that a log P value around 2.5 should be the optimal lipophilicity value for the activity of THPP-containing compounds. PMID- 12724038 TI - The polysaccharide from Tamarindus indica (TS-polysaccharide) protects cultured corneal-derived cells (SIRC cells) from ultraviolet rays. AB - The aim of this work was to investigate the possible protective effect of a new viscosising agent, TS-polysaccharide, on corneal-derived cells (SIRC) exposed to ultraviolet-B rays. To verify this, SIRC cells were first exposed, in the absence or in the presence of TS-polysaccharide (1% w/v), for 9 s at the UV-B source and then post-incubated for 45 min at 37 degrees C. After this period the hydrogen peroxide (H(2)O(2)) accumulated in the medium and the concentration of 8-hydroxy 2'-deoxy-guanosine (8-OHdG) in cell DNA was measured. In addition, the amount of (3)H-methyl-thymidine incorporated in cellular DNA was evaluated after 18 h from irradiation. Our results show that cells exposed to UV-B rays accumulate H(2)O(2), and have higher levels of 8OHdG and a lower amount of (3)H-methyl thymidine incorporated in DNA than control cells. In the presence of TS polysaccharide, the H(2)O(2) and 8-OHdG accumulation, and the (3)H-methyl thymidine incorporation were significantly reduced with respect to the values measured in cells exposed in the absence of the polysaccharide. We propose a protective role of the polysaccharide in reducing UV-B derived DNA damage to eye cells. This finding could be of some clinical importance when the polysaccharide is used as a delivery system for ophthalmic preparations. PMID- 12724039 TI - Thiorphan enhances bradykinin-induced vascular relaxation in hypoxic/hyperkalaemic porcine coronary artery. AB - Relaxation induced by bradykinin is diminished by hypoxia in epicardial coronary arteries. The bradykinin-degrading enzyme, neutral endopeptidase (NEP, EC.3.4.24.11), is a potential target for coronary artery vasodilators. In this study, we examined the effect of thiorphan, an inhibitor of NEP, on the tone of porcine isolated coronary artery under hypoxic conditions. Endothelium-intact porcine isolated coronary artery rings were isometrically contracted with a prostaglandin F(2alpha) analogue (U46619, 0.75 microM) and potassium chloride (KCl, 30 mM), and relaxed with bradykinin (1-1000 nM) under normoxic (partial pressure of oxygen, pO(2) approximately 90-100 mmHg) and moderately hypoxic (pO(2) approximately 50-60 mmHg) conditions. Experiments were performed to study the effects of 30 min pre-treatment with the NEP-inhibitor, thiorphan (10 microM), both at physiological and at low pO(2)s. Hypoxia inhibited the bradykinin-induced relaxation in porcine epicardial coronary arteries. In normoxia, thiorphan significantly enhanced the decrease of coronary tone produced by bradykinin (1-10 nM) when U46619 was used as contractile agent. Under hypoxic conditions, in U46619 contracture, thiorphan did not influence, but in KCl contracture it enhanced the magnitude of relaxations induced by bradykinin. In the absence of bradykinin, thiorphan had no significant effect on the basal, KCl- and U46619-elevated tones and on the hypoxia-induced decrease of coronary artery tone. Inhibition of NEP-enzyme activity may effectively improve the relaxing capacity of epicardial coronary arteries under hypoxic/hyperkalemic conditions. This effect could be potentially utilized when the endothelial function and relaxation of the coronary arteries are impaired under clinical conditions. PMID- 12724040 TI - Butein suppresses myofibroblastic differentiation of rat hepatic stellate cells in primary culture. AB - Hepatic stellate cells play a key role in the pathogenesis of hepatic fibrosis. In this study, we investigate the inhibitory effect of butein on the activation and proliferation of rat primary cultured hepatic stellate cells. Possible cytotoxic effects were measured on stellate cells and hepatocytes using the 3 (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The effects of butein on the production of collagen and smooth muscle alpha-actin proteins were examined at the same concentration, by western blot. The effects of butein on alpha1(I) collagen, tissue inhibitor of metalloproteinase-1, and metalloproteinase-13 gene expression in activated stellate cells were investigated by measuring mRNA levels using reverse transcription polymerase chain reaction. The effect of butein on DNA synthesis was also determined. Butein, at a concentration of 1 microg mL(-1), reduced DNA synthesis without affecting cell viability, and downregulated smooth muscle alpha-actin and type-I collagen expression, and alpha1(I) collagen and tissue inhibitor of metalloproteinase-1 mRNA expression, while treatment with butein induced metalloproteinase-13 mRNA expression. These findings suggest that butein is a potent inhibitor of stellate cell transformation. PMID- 12724041 TI - Quercetin, a phytoestrogen and dietary flavonoid, activates different membrane bound guanylate cyclase isoforms in LLC-PK1 and PC12 cells. AB - Accumulated evidence suggests that quercetin, a dietary flavonoid, has beneficial effects in protection against cardiovascular diseases and in the inhibition of tumour growth. We have recently shown that antioxidants such as 17beta-estradiol, resveratrol, dithiothreitol and vitamin C activate membrane-bound guanylate cyclase GC-A, a receptor for atrial natriuretic factor (ANF). Since quercetin is a phytoestrogen and potent antioxidant, it is possible that it may activate GC-A or other guanylate cyclase isoforms. We examined whether quercetin activates GC-A or GC-B (the receptor for C-type natriuretic peptide, CNP) in PC12 and porcine kidney proximal tubular LLC-PK1 cells. The results showed that quercetin activated a guanylate cyclase isoform in both cell types. Quercetin inhibited CNP stimulated GC-B activity, but had little effect on ANF-stimulated GC-A activity in PC12 cells, suggesting that quercetin mainly activates GC-B in PC12 cells. In contrast, CNP had no effect on guanylate cyclase activity in LLC-PK1 cells, indicating that GC-B is not expressed in LLC-PK1 cells. Furthermore, quercetin had a small effect on ANF-stimulated GC-A activity and had no effect on soluble guanylate cyclase activity in LLC-PK1 cells, suggesting that quercetin does not activate GC-A, GC-B or soluble guanylate cyclase in LLC-PK1 cells. However, quercetin did stimulate membrane-bound guanylate cyclase activity in LLC-PK1 cell membranes. These results indicate that quercetin activates the GC-B isoform in PC12 cells, but activates an unknown membrane-bound guanylate cyclase isoform in LLC-PK1 cells. PMID- 12724042 TI - Endothelium-dependent and -independent vasodilator effects of eugenol in the rat mesenteric vascular bed. AB - The possible involvement of the endothelium in the vasodilator action of eugenol was investigated in the mesenteric vascular bed (MVB) of the rat. Bolus injections of eugenol (0.2, 2 and 20 micromol) and acetylcholine (ACh; 10, 30 and 100 pmol) induced dose-dependent vasodilator responses in noradrenaline precontracted beds that were partially inhibited by pretreatment of the MVB with deoxycholate (1 mg mL(-1)) to remove the endothelium (approximately 14% and approximately 30% of the control response remaining at the lowest doses of ACh and eugenol, respectively). The vasodilator effect of glyceryl trinitrate (1 micromol) was unaltered by deoxycholate. In the presence of either N(omega)-nitro L-arginine methyl ester (300 microM) or tetraethylammonium (1 mM)the response to ACh was partially reduced, whereas eugenol-induced vasodilation was unaffected. Similarly the vasodilator effect of eugenol was not inhibited by indometacin (3 microM). Under calcium-free conditions the vasoconstrictor response elicited by bolus injections of noradrenaline (10 nmol) was dose-dependently and completely inhibited by eugenol (0.1-1 mM). Additionally, the pressor effects of bolus injections of calcium chloride in potassium-depolarized MVBs were greatly reduced in the presence of eugenol (0.1 mM), with a maximal reduction of approximately 71% of the control response. Our data showed that eugenol induced dose-dependent, reversible vasodilator responses in the rat MVB, that were partially dependent on the endothelium, although apparently independent of nitric oxide, endothelium derived hyperpolarizing factor or prostacyclin. Furthermore, an endothelium independent intracellular site of action seemed likely to participate in its smooth muscle relaxant properties. PMID- 12724043 TI - Changes of materials that scavenge 1,1-diphenyl-2-picrylhydrazyl radicals in plasma by per-oral administration of Kampo medicine, Ninjin-yoei-to in rats. AB - The Kampo medicine, Ninjin-yoei-to, scavenged 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals in a dose-dependent fashion as did ascorbic acid and alpha tocopherol. Ninjin-yoei-to, which is composed of 12 herbs, had a potent DPPH radical scavenging ability. We investigated the transition of the materials that scavenge DPPH radicals in plasma after oral administration of Ninjin-yoei-to to rats. When 1.0 g kg(-1) Ninjin-yoei-to was administered, the DPPH radical scavenging ability increased at 30 min and biphasic peaks were observed at 2 h and at 10 h. From the response-time profile, kinetic parameters including values for K(a) (absorption rate constant), t(max) (peak concentration time), t(1/2) (half-life) and MRT (mean residence time) of the radical scavenging ability in plasma could be calculated for DPPH radicals. K(a) values were 0.53 +/- 0.03 and 0.36 +/- 0.07 h, t(max) values were 2.1 +/- 1.04 and 8.56 +/- 2.69 h, t(1/2) values were 1.60 +/- 0.12 and 3.39 +/- 1.72 h, and MRT values were 4.14 +/- 1.59 and 8.18 +/- 2.55 h, respectively. These parameters calculated from the antioxidation dynamics were considered to offer a very meaningful procedure for examining the effects of Ninjin-yoei-to. PMID- 12724044 TI - Dual effect of agmatine in the bisected rat vas deferens. AB - The functional effects of the amine agmatine, the putative endogenous ligand for alpha(2)-adrenoceptors and imidazoline receptors, in rat vas deferens were investigated by using the epididymal and prostatic portions. Tissues were contracted by electrical stimulation or by exogenous drugs. In electrically stimulated portions, agmatine caused a dual effect on contractions. In the epididymal portion an inhibition on twitch contractions was observed, which was partially antagonised by idazoxan and yohimbine, indicating the involvement of at least a presynaptic alpha(2)-adrenoceptor-mediated mechanism, without the interference of imidazoline receptors. In the prostatic portion, agmatine enhanced the amplitude of twitches. In contractions induced by exogenous drugs, agmatine potentiated, only in the prostatic segment, the effects of noradrenaline (norepinephrine) or ATP; it also enhanced the effect of low concentrations of KCl and blocked the maximum effect of the higher concentrations. Effects induced by agmatine on the exogenous ATP in the prostatic portion were blocked by cromakalim, suggesting a blocking action on the postsynaptic K(+) channels, which explains, in part, the potentiation of the twitch amplitude. It was concluded that agmatine interferes with sympathetic neurotransmission, but the physiological relevance of this needs to be better understood because of the high doses employed to induce its effects. PMID- 12724045 TI - Apparent mechanism-based inhibition of human CYP3A in-vitro by lopinavir. AB - The influence of the viral protease inhibitor lopinavir on the activity of six human cytochrome P450 (CYP) enzymes was evaluated in a model system using human liver microsomes. Column chromatography methodology was developed to separate lopinavir from ritonavir starting from the commercially available lopinavir ritonavir combination dosage form. Lopinavir produced negligible or weak inhibition of human CYP1A2, 2B6, 2C9, 2C19 and 2D6. However, lopinavir was an inhibitor of CYP3A. At 250 microM triazolam (the CYP3A index substrate), the mean (+/- s.e., n = 4) IC50 versus triazolam alpha-hydroxylation (where IC50 is the concentration producing a 50% decrement in reaction velocity) was 7.3 (+/- 0.5) microM. Pre-incubation of lopinavir with microsomes prior to addition of triazolam yielded a significantly lower IC50 of 4.1 (+/- 0.5) microM. This is consistent with mechanism-based inhibition of human CYP3A by lopinavir. Although lopinavir is less potent than ritonavir as an inhibitor of CYP3A, lopinavir is nonetheless likely to contribute to net CYP3A inhibition in-vivo during treatment with the lopinavir-ritonavir combination. PMID- 12724046 TI - Effects of folic acid and lamotrigine therapy in some rodent models of epilepsy and behaviour. AB - It has been suggested that a folic acid (FA) deficiency induced by antiepileptic drugs might be the basis for the neuropsychiatric toxicity associated with these drugs. In the present study, lamotrigine (LTG), one of the newer antiepileptic drugs, was evaluated for its effect on epilepsy, mood and memory in mice. Further, the effect of the addition of FA to LTG therapy was also investigated. The increasing current electroshock seizure test was used to evaluate the anticonvulsant effect of drugs, while the forced swimming test (FST) and spontaneous alternation behaviour (SAB) models were employed for assessing the effects on mood and memory, respectively. LTG exhibited a dose-dependent increase in seizure threshold, whereas FA did not have any effect. LTG did not affect, whereas FA decreased, behavioural depression in the FST in mice. Neither LTG nor FA affected memory scores in the SAB test. The combination of LTG and FA significantly reduced depression while enhancing the effects on memory and seizure threshold. The present observations have confirmed the antiepileptic action of LTG in yet another rodent model of epilepsy. Further, the results clearly demonstrate the additional benefits on epilepsy, mood and memory brought about by the inclusion of FA in the LTG regimen. PMID- 12724047 TI - Comparison of suppressive potency between azathioprine and 6-mercaptopurine against mitogen-induced blastogenesis of human peripheral blood mononuclear cells in-vitro. AB - Azathioprine (AZ) is a prodrug of 6-mercaptopurine (6-MP), but little is known about the relative suppressive efficacy of these agents against blastogenesis of human peripheral blood mononuclear cells (PBMCs) in-vitro. We compared the pharmacological efficacy of AZ and 6-MP against T cell mitogen-induced blastogenesis of PBMCs in-vitro. PBMCs were obtained from seven healthy subjects. The potency of AZ and 6-MP to suppress PBMC-blastogenesis in-vitro was compared using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay procedures. Production of 6-MP from AZ in PBMC culture was examined by high performance liquid chromatography. Both AZ and 6-MP dose-dependently suppressed PBMC blastogenesis. Mean +/- s.d. IC50 (concentration of drug that gave 50% inhibition) values for AZ and 6-MP were 230.4 +/- 231.3 and 149.5 +/- 124.9 nM, respectively. Thus, the potencies of AZ and 6-MP to suppress PBMC blastogenesis were not significantly different. A significant correlation was observed between the IC50 values of AZ and 6-MP (P < 0.01, n = 6). After incubation of PBMCs with 100 microM AZ for 4 days, 35.3-92.5 microM 6-MP was liberated into the culture medium. The ratio of 6-MP liberation from AZ was influenced by the presence of PBMCs, but not by the medium or fetal bovine serum. The results suggest that the suppressive potency of the prodrug AZ and its converted form 6-MP against blastogenesis of human PBMCs in-vitro is similar, although PBMCs appeared to convert AZ to 6-MP. AZ is suggested to be effective after conversion to 6-MP to express immunosuppressive efficacy in-vitro. PMID- 12724048 TI - Improvement of (+)-catechin inhibitory activity on human PMN respiratory burst by (+)-3-O-propionyl and (-)-3-O-valeryl substitution. AB - Catechins and their derivatives are abundant flavanols in the plant kingdom. Usually, catechin activity correlates with chemical structure. We hypothesized that by adding hydrophobic groups to the native catechin, we could ameliorate penetration of the cell and make the derivatives more active than native molecule in inhibiting polymorphonuclear leucocyte (PMN) oxidative burst. This study was designed to compare the antioxidant activity of native catechin with that of (+) 3-propionylcatechin and (-)-3-Ovalerylcatechin esters by two cell-free colorimetric methods and by their effects on whole blood leucocytes as well as on isolated PMN chemiluminescence activity. The results showed that the colorimetric methods did not detect differences between catechins. On the contrary, cellular chemiluminescence studies showed that light emission by resting, as well as by phorbol myristate acetate (PMA)-stimulated PMNs and whole blood leucocytes was inhibited by catechin esters more intensively than native catechin. The compartmental chemiluminescence evaluation showed that the extracellular activity was similar with all catechins, while the intracellular activity was higher with esters. PMN pre-incubation, with catechins at various times before stimulation with PMA, enhanced the inhibitory activity of all compounds. Since the esterification with propionic or valeric acid increased the lipophilicity of (+) catechin, we hypothesized that native and esterified catechins have different intracellular availability and therefore differ in effectiveness. An ancillary result obtained is that a single approach, chemical or cellular, is not sufficient to evaluate overall antioxidant activity in biological sytems. The results indicate that modified catechins may be very intriguing as possible future leucocyte modulating drugs, with possible applications in vascular and inflammatory diseases. PMID- 12724049 TI - The Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial (ALLHAT). Major outomes in high-risk hypertensive patients randomized to angiotensin-converting enzyme inhibitor or calcium channel blocker vs diuretic. PMID- 12724050 TI - Optimizing target-organ protection in patients with diabetes mellitus: angiotensin-converting enzyme inhibitors or angiotensin receptor blockers? AB - High blood pressure in the setting of type 1 and type 2 diabetes is commonly associated with the earlier development of target-organ damage, including cardiovascular and cerebrovascular disease and progressive renal insufficiency. The major goal of treating high blood pressure in this population is to prevent or reduce the likelihood of target-organ damage. The treatment goal for high blood pressure, therefore, has to be defined based on optimal means of preventing cardiovascular and renal events. The reduction of high blood pressure with pharmacologic therapy is associated with reduction of cardiovascular events, renal disease, and associated mortality. However, many questions remain. Some of the basic and important questions include the following: What should be the goal of treated blood pressure in the diabetic, and are there preferred agents that should be used in the hypertensive diabetic population? How do angiotensin converting enzyme inhibitors and angiotensin receptor blockers fit in? Are there advantages of one class over the other? The goal of this review is to summarize the recent clinical trial findings and try to provide recommendations based on the evidence of these trials to help the clinician better choose blood pressure goals and treatment strategies in the diabetic population. PMID- 12724051 TI - Potential mechanisms of impaired endothelial function in arterial hypertension and hypercholesterolemia. AB - This review focuses on the role of impaired endothelial function for the development of atherosclerosis in human arterial hypertension and hypercholesterolemia in vivo. Potential mechanisms underlying impaired endothelial function and decreased bioavailability of nitric oxide under these clinical conditions are discussed. It further addresses therapeutic strategies aimed at improving the bioavailability of nitric oxide in these patients. The overall conclusion is that the bioavailability of nitric oxide is probably impaired, not by a single defect, but by various mechanisms affecting nitric oxide synthesis as well as nitric oxide breakdown. In both diseases increased superoxide anion production and oxidative stress represent a major mechanism. Decreased bioavailability of nitric oxide not only impairs endothelium-dependent vasodilation, but also activates other mechanisms that play an important role in the pathogenesis of atherosclerosis. Thus, therapeutic strategies should aim to restore bioavailability of nitric oxide, which has been demonstrated for lipid lowering therapy in hypercholesterolemia and blood pressure control in hypertension. In addition, antioxidative strategies will represent a major therapeutic tool against atherosclerotic diseases in the future. Statins and blockers of the renin-angiotensin system seem to have such antioxidative effects independent from their effects on lipid profiles or blood pressure control. PMID- 12724053 TI - Oxidative stress and heart disease: cardiac dysfunction, nutrition, and gene therapy. AB - Oxidative stress is defined as the imbalance between the generation of reactive oxygen species and antioxidant defense mechanisms. The cardiovascular system is a major target for reactive oxygen species. Cardiomyocytes and the vasculature of the heart can be severely damaged as a result of oxidative stress. In this paper, we discuss recent findings with respect to the role of oxidative stress in heart disease. The efficacies of treatments with vitamins and wine-derived compounds, as well as innovative gene therapeutic experiments that may potentially alleviate oxidative stress-induced disease, are also discussed. PMID- 12724052 TI - Microalbuminuria in hypertension. AB - A body of evidence indicates that microalbuminuria is a well-recognized marker of cardiovascular complications and increased cardiovascular risk in hypertension. However, the prognostic significance of microalbuminuria remains controversial because only the results of a few prospective studies performed in small groups of hypertensive subjects without diabetes mellitus are available. Several factors can affect the prevalence of microalbuminuria in hypertension including age, sex, race, severity of the disease, and concomitant risk factors. This accounts for the large differences in the prevalence of microalbuminuria that can be found in the literature, with prevalence rates going from a low of 4.7% to a high of 46%. The main determinant of albumin excretion rate in subjects with mild hypertension and no cardiovascular complications seems to be the hemodynamic load, whereas in subjects with more severe hypertension and associated target organ damage, the augmented urinary albumin leak is probably the consequence of glomerular damage. Inhibition of the renin-angiotensin system with angiotensin-converting enzyme inhibitors or angiotensin II receptor antagonists is particularly effective at reducing the albumin excretion rate, but whether these classes of drugs are more beneficial in patients with microalbuminuria remains to be determined. There is general consensus that evaluation of microalbuminuria is useful for the assessment of overall cardiovascular risk in hypertension, since albumin excretion rate appears to be a cost-effective way to identify patients at higher risk for whom additional preventive and therapeutic measures are advisable. PMID- 12724054 TI - Are beta-blockers efficacious as first-line therapy for hypertension in the elderly? AB - The aim of this paper is to assess antihypertensive efficacy of b-blockers and their effects on cardiovascular morbidity and mortality and all-cause mortality in elderly patients with hypertension. Randomized placebo-controlled trials, trials with untreated controls, and trials comparing antihypertensive drugs were selected from the literature. The relative risk reduction of primary endpoints and 95% confidence intervals were calculated. There were six trials in which elderly patients were treated with beta-blockers for hypertension (three trials were placebo-controlled, one trial had untreated controls, and two trials compared antihypertensive drugs). There was no study with monotherapy of beta blockers. In combination with diuretics, beta-blockers were superior to placebo and untreated controls in preventing cardiovascular events, especially strokes, but there was no superiority of beta-blockers to angiotensin-converting enzyme inhibitors and calcium antagonists in preventing cardiovascular morbidity and mortality and total mortality in elderly patients with hypertension. Beta blockers are only efficacious in combination with diuretics in preventing cardiovascular morbidity and mortality in elderly patients with hypertension. PMID- 12724055 TI - The renin-angiotensin system in blacks: active, passive, or what? AB - Plasma renin activity in blacks has been consistently reported to be lower than in whites. Many mechanisms for the low plasma renin activity have been proposed, including volume status, renal sodium handling, and the reduction of renin release. The status of the RAS is paramount in the regulation of salt and water balance and its implications in disease processes such as hypertension and renal failure. In this review, we present data to suggest that low systemic plasma renin activity in blacks may not be the primary abnormality, but rather the reflection of an overactive RAS at the tissue level in the kidney. PMID- 12724056 TI - Are low target blood pressure goals justified in persons with diabetes mellitus? AB - Hypertension is common in patients with diabetes and is a major risk factor for development and progression of the macro- and microvascular complications seen in diabetes. The Joint National Committee VI recommendation for goal blood pressure is less than 130/85 mm Hg in diabetics--a more aggressive target than in nondiabetic patients. Data over the past decade support these aggressive goals, especially for cardiovascular and renal outcomes and overall mortality. In addition, in diabetics, blood pressure appears to be a continuous risk factor for these outcomes without evidence of a J-point effect. While these goals are rarely obtained in diabetic patients, studies demonstrate that they are achievable with attention to detail and use of multiple antihypertensive agents. PMID- 12724057 TI - Effects of hormone replacement therapy on the sympathetic nervous system and blood pressure. AB - Hypertension is a major health problem that significantly contributes to heart disease and stroke. While most studies of hypertension have focused on men, women also experience significant hypertension-related morbidity and mortality. However, the incidence of hypertension and cardiovascular disease is significantly lower in premenopausal women compared with men until the onset of menopause, at which time cardiovascular disease incidence increases dramatically in women and eventually approaches that in men. These observations indicate that the loss of estrogen contributes to menopause-related increases in blood pressure and cardiovascular disease, and suggest that the use of estrogen hormone replacement therapy could decrease the incidence of cardiovascular disease in postmenopausal women. However, new findings from the Women's Health Initiative study suggest that estrogen therapy has few positive benefits and some significant negative effects on the health of postmenopausal women, and these data have caused many to abandon long-term estrogen replacement therapy. Conversely, numerous clinical and basic research studies indicate that estrogen replacement therapy beneficially reduces blood pressure, thereby decreasing the incidence of hypertension and cardiovascular disease. Further, several of these studies suggest that one means by which estrogen lowers blood pressure is by decreasing sympathetic nervous system activity. This review examines the evidence supporting estrogen's ability to modulate sympathetic nervous system tone and thereby decrease arterial pressure. PMID- 12724058 TI - Insulin resistance and the sympathetic nervous system. AB - The obesity epidemic is driving metabolic (insulin resistance) syndrome-related health problems including an approximately threefold increased coronary heart disease risk. Sympathetic hyperfunction may participate in the pathogenesis and complications of the metabolic syndrome including higher blood pressure, a more active renin-angiotensin system, insulin resistance, faster heart rates, and excess cardiovascular disease including sudden death. Possible factors augmenting sympathetic activation in the metabolic syndrome include alterations of insulin, leptin, nonesterified fatty acids (NEFAs), cytokines, tri-iodothyronine, eicosanoids, sleep apnea, nitric oxide, endorphins, and neuropeptide Y. Of note, high plasma NEFAs are a risk factor for hypertension and sudden death. In short term human studies, NEFAs can raise blood pressure, heart rate, and a(1) adrenoceptor vasoreactivity, while reducing baroreflex sensitivity, endothelium dependent vasodilatation, and vascular compliance. Efforts to further identify the mechanisms and consequences of sympathetic dysfunction in the metabolic syndrome may provide insights for therapeutic advances to ameliorate the excess cardiovascular risk and adverse outcomes. PMID- 12724059 TI - Hypertension and cognitive function: pathophysiologic effects of hypertension on the brain. AB - Accumulating evidence supports a causal role of hypertension for cognitive decline above and beyond its relationship to frank stroke. Hypertension associated pathologic changes in the brain and its vasculature include vascular remodeling, impaired cerebral autoregulation, cerebral microbleeds, white matter lesions, unrecognized lacunar infarcts, and Alzheimer-like changes such as amyloid angiopathy and cerebral atrophy. White matter lesions and retinal vascular changes, both of which can be imaged noninvasively, may reveal the general condition of the cerebral vasculature or the presence of arteriosclerotic or hypertensive encephalopathy. These noninvasive indicators may also identify a subgroup in whom infarct prevention, particularly via blood pressure reduction, is of paramount importance. Optimal control of blood pressure on a population wide basis, but particularly in those prone to cognitive loss, is thus a critically important but as yet elusive goal that should be fully exploited for its potential to reduce future cognitive impairment. PMID- 12724061 TI - The role of the sympathetic nervous system in linking obesity with hypertension in white versus black Americans. AB - Several previous studies confirmed that obesity is a major risk factor for the development of cardiovascular diseases, including hypertension. A large number of clinical studies considered the role of the sympathetic nervous system in linking obesity with hypertension, and there is substantial evidence that human obesity is characterized by defects in sympathetic cardiovascular control. The association of obesity with hypertension has been well documented in most racial, ethnic, and socioeconomic groups. Ethnicity may be an important factor to consider since sympathetic nervous system activity, and the propensity for obesity and hypertension, all differ substantially among different populations. Obesity is actually accompanied by increased sympathetic nerve discharge to skeletal muscles, a main site for energy expenditure. Adiposity-related sympathetic overactivity is a compensatory mechanism to burn fat and decrease weight gain, but at the cost of increased sympathetic discharge to the peripheral vasculature, which could predispose to hypertension. Thus, sympathetic nervous system activity is important in the development and maintenance of obesity related hypertension in different racial and ethnic populations, including white and black Americans. PMID- 12724060 TI - Brainstem mechanisms of hypertension: role of the rostral ventrolateral medulla. AB - The central nervous system plays a key role in the regulation of cardiovascular function, and alterations in the central neural mechanisms that control blood pressure may underlie the vast majority of cases of primary hypertension. The well-studied baroreceptor reflex powerfully regulates arterial pressure, though its involvement in the pathogenesis of chronic hypertension is likely to be only of minor importance. Supraspinal maintenance of sympathetic vasomotor outflow appears to emanate from neurons in the rostral ventrolateral medulla, and the tonic drive exerted on sympathetic vasomotor activity by the rostral ventrolateral medulla appears to be increased in several animal models of hypertension. In particular, the excitation of the rostral ventrolateral medulla by excitatory amino acid neurotransmitters and by stimulation of AT(1) angiotensin receptors appears to be increased in experimental hypertension. The current data support the view that neurogenic hypertension is mediated by increased excitatory drive of rostral ventrolateral medulla sympathoexcitatory neurons. PMID- 12724063 TI - [Circadian blood pressure variation in normal pregnancy, gestational hypertension, and preeclampsia]. AB - BACKGROUND AND OBJECTIVE: Changes in circadian variation of blood pressure could be used either to predict preeclampsia or to assess its severity. With the objective of identifying potential differences in blood pressure at the early stages of pregnancy, we examined and compared the characteristics of circadian variability in blood pressure in healthy and complicated pregnant women who were systematically monitored throughout gestation. SUBJECTS AND METHOD: We analyzed 2,014 blood pressure series sampled through ambulatory monitoring for 48 hours once every 4 weeks from the first obstetric visit until delivery. The study included 205 women with uncomplicated pregnancy, 92 with gestational hypertension and 31 with preeclampsia. The circadian pattern of blood pressure variation for each group and trimester of gestation was established by means of a population multiple-components analysis. RESULTS: Differences in the 24-hour mean between healthy and complicated pregnancies were highly significant in all trimesters (p < 0.001), with values of 15.1 and 9.1 mmHg for systolic and diastolic blood presure, respectively, in the third trimester of pregnancy. The 24-hour mean of systolic/diastolic blood pressure for complicated pregnancies was always below 120/72 mmHg. Results further indicated similar circadian characteristics between gestational hypertension and preeclampsia in the first trimester of pregnancy. The difference between these two groups in the 24-hour mean was significant in the second trimester for systolic (3 mmHg; p = 0.002) but not diastolic blood pressure (0.9 mmHg; p = 0.230). In the third trimester, the difference between gestational hypertension and preeclampsia was significant for both variables (5.4 and 3.7 mmHg for systolic and diastolic blood pressure, respectively; p < 0.001). CONCLUSIONS: The differences in blood pressure between healthy and complicated pregnancies, which are observed as early as the first trimester of pregnancy, are detected when both systolic and diastolic blood pressure measurements in women with a late diagnosis of gestational hypertension or preeclampsia fall within accepted ranges of normotension. These differences offer new end points that may lead to an early identification of hypertensive complications in pregnancy as well as to the establishment of prophylactic interventions. PMID- 12724062 TI - Genetics of the kidney and hypertension. AB - Success in the search for genes that cause or contribute to hypertension susceptibility has been limited to a few rare Mendelian forms of hypertension (glucocorticoid remediable aldosteronism, apparent mineralocorticoid excess, and Liddle's syndrome). Our well-reasoned efforts to assess candidate genes in critical pathways known to be involved in blood pressure regulation have not been as productive in complex genetic cases of hypertension. These cases involve both genetic and environmental determinants. The most frequently used approach to the identification of hypertension genes involves genetic association studies, which are population based and compare cases and controls. Linkage analyses are also used but require family data. While much effort is spent identifying new markers and candidate genes, it is important to periodically determine which findings of linkage or association are confirmed in order to advance our quest to identify hypertension genes. In this review, the status of the assessment of the HSD11B2 gene is reviewed. In addition, data supporting the need to assess the mitochondrial genome, the other human genome, in hypertension susceptibility are reviewed. PMID- 12724064 TI - [Control of blood pressure in diabetic patients in primary care setting. DIAPA study]. AB - BACKGROUND AND OBJECTIVE: We aimed to assess the blood pressure (BP) control in patients with diabetes mellitus (DM) treated in primary care (PC) and to establish the factors associated with a suboptimal control of BP. PATIENTS AND METHOD: This was a multicenter, transverse study. Ninety investigators included 875 patients with DM (57.8% women), with a mean (SD) age of 64 (11.8) years. BP was measured according to the Sixth Report of Joint National Committee (JNC-VI) and Word Health Organization/International Society Hypertension (WHO/ISH) recommendations, calculating the arithmetic mean of three consecutive measurements. Patients with a previous established diagnosis were considered to be hypertensive, and an optimal control was considered when BP values were lower than 130/85 mmHg. RESULTS: 66.7% (n = 583) patients with DM were previously diagnosed of hypertension (HT); 86.3% (CI 95%: 83.0-89.1) of those with type 2 DM and 87.8% (CI95%: 70.9-96.0) of those with type 1 DM did not have an optimal control of BP at the visit (BP >= 130 and/or 85 mmHg). 56.8% (CI 95%: 50.1-63.3) of those with type 2 DM and 23.8% (CI 95%: 12.6-39.8) of those with type 1 DM with unknown HT had a BP >= 130 and/or 85 mmHg at the visit. The median of antihypertensive drugs used was 1 (1-2). In a multivariate analysis, age, higher body mass index (BMI), higher cholesterol levels and uric acid were associated with a worse BP control (p < 0.001). CONCLUSIONS: In our study, 66.7% of patients with DM attended in PC had known HT and only 13.6% had an optimal BP control. Diabetic patients with HT were undertreated, with a median of one antihypertensive drug. PMID- 12724065 TI - [Analysis of temporal evolution in alcohol consumption during pregnancy in the Spanish autonomic communities]. AB - BACKGROUND AND OBJECTIVE: The fetal alcohol syndrome (FAS) is considered as the main preventable cause of congenital defects and mental deficiency. We analyzed whether the consumption of alcohol during gestation has changed in Spain. SUBJECTS AND METHOD: We analyzed the evolution over the last 24 years, as well as the geographical distribution according to Spanish regions, of maternal alcohol consumption during pregnancy. We used data from the ECEMC, a registry which includes 1,820,862 liveborns, of whom 30,836 were malformed. A similar number of healthy controls was collected. We established 5 increasing levels of alcohol consumption, for sporadic and diary intakes, and 3 levels of maternal scholarship. RESULTS: Except for the lowest and highest intakes, alcohol consumption levels showed a decreasing trend over the time. These tendencies were similar in almost all Spanish regions. When comparing the intake between regions, the differences were statistically significant. There was a correlation between the maternal scholarship and the alcohol intake, showing that the higher the education the lower the amount of alcohol consumption. CONCLUSIONS: The awareness of the alcohol effects is related to the maternal cultural level. Maternal consumption of alcohol varies across Spanish regions, though its trends are similar. PMID- 12724066 TI - [Influenza vaccine in pregnant women]. PMID- 12724067 TI - [Refractoriness to platelet transfusions]. PMID- 12724068 TI - [Cyclooxygenase-2 and gastroduodenal lesions. Any relationship with Helicobacter pylori ? A systematic review]. PMID- 12724069 TI - [Splenic infartion and subcapsular haematoma in the course of chronic pancreatitis]. PMID- 12724070 TI - [Chronicity of hypercoagulative state: a favourable evidence for postoperative prolongation of antithrombotic prophylaxis]. PMID- 12724071 TI - [Cardiovascular risk factors and healthy habits in the pediatric age group]. PMID- 12724072 TI - [Cardiovascular risk factors in children and teenagers aged 6-18 years old from Medellin (Colombia)]. AB - OBJECTIVE: To evaluate exposure to the major risk factors for cardiovascular disease in children from Medellin according to age, sex, type of school, and socioeconomic status. METHOD: We performed a descriptive study in 2611 children aged 6-18 years old from the city of Medellin in Colombia. Lipid profile, blood pressure, body mass index, diet, exercise, alcohol intake, and smoking were evaluated. RESULTS: Forty-six percent of the children drank alcohol, 8.7 % smoked and 50 % were physically inactive. Fat and carbohydrate intake was high in 48 % and 47 %, respectively. A total of 9.3 % of the children were overweight and 4.6 % were obese. Systolic and diastolic blood pressure were high in 1.3 % and 3.9 %, respectively. Mean high-density lipoprotein cholesterol (HDL-C) values were lower than in other populations, while triglyceride values were higher. Differences in serum lipid concentrations were found according to age and sex. The prevalence of risk factors according to the National Cholesterol Education Program criteria were: 19.1 % for HDL-C, 17.1 % for triglycerides, 17.0 % for low-density lipoprotein cholesterol (LDL-C), 13.5 % for total cholesterol (TC) and 22.9 % for TC/HDL-C. CONCLUSIONS: In children from the city of Medellin, the most prevalent cardiovascular risk factors were related to lifestyle. Mean plasma lipid concentrations varied according to age and sex. The prevalence of overweight was higher than in other populations in Colombia but was lower than that reported for other countries. PMID- 12724073 TI - [Nutritional status and immunoglobulin and cytokine concentrations in children with malaria]. AB - OBJECTIVES: To relate nutritional status and concentrations of immunoglobulins and cytokines in children with malaria from two areas with different risk of malaria transmission. METHODS: We performed a descriptive, cross-sectional study comparing children aged 4-11 years old from two areas with different risk of malaria transmission in Colombia. The sample consisted of 66 children from El Bagre and Zaragoza (high transmission area) and 62 children from Turbo (low transmission area). To determine the risk of undernutrition, height/weight, age/height and weight/age indexes were calculated, and serum concentrations of interleukin-10 (IL-10), tumor necrosis factor-alpha (TNF-alpha), total IgE and malaria-specific IgE were measured. RESULTS: In the high transmission area, concentrations of total and specific IgE and of TNF-alpha were significantly higher. In both areas, the values obtained for total IgE (84 %), specific-IgE (32 %), TNF-alpha (72 %) and IL-10 (84 %) were higher than standard values. Anthropometric indicators revealed acute undernutrition (wasting) in 33 %, chronic undernutrition (stunting) in 52 %, and global undernutrition in 56 % of the population. CONCLUSIONS: Malaria and protein-energy malnutrition were highly prevalent in both areas. In children from the low transmission area, stunting was significantly greater. In the high transmission area, the mean total IgE was twice that found in the low transmission area and no association with nutritional status was observed. Levels of specific IgE did not differ according to the species of Plasmodium infection. PMID- 12724074 TI - [Toxocariasis: clinical and laboratory features in 54 patients]. AB - OBJECTIVE: To describe the clinical and laboratory findings in children with toxocariasis. METHODS: Fifty-four children with reactive serology to Toxocara determined by ELISA were prospectively identified between January 1998 and September 2000. The patients were divided into three groups: asymptomatic children (n 24), those with visceral larva migrans (n 16) and those with ocular larva migrans (n 14). Age, serology titers, and eosinophil count at diagnosis were compared among the groups. The patients received treatment with albendazole 10-15 mg/kg/day for 15 days or thiabendazole 25 mg/kg/day in two series of 7 days. RESULTS: The clinical features were as follows: 24 children (44.4 %) were asymptomatic, pneumonitis was found in 9 (16.7 %), hepatomegaly in 6 (11.1 %), acute posterior uveitis in 5 (9.3 %), strabismus in 5 (9.3 %), leukocoria in 4 (7.4 %), fever in 3 (5.6 %). There was 1 case of keratitis, 1 of cataracts, 1 of myocarditis and 1 case of pneumonia with pleural effusion. Some patients showed more than one clinical feature. Four children experienced loss of vision in the affected eye. No differences in age or serology titers were found among the groups. Eosinophil count was lower in the group with ocular larva migrans than in the other groups (p < 0.001). Children with active disease showed clinical improvement and a 70.4 % decrease in eosinophilic count one year after treatment. Serological titers showed an unpredictable pattern during the follow-up. CONCLUSIONS: Most of the infected children were asymptomatic. In the post treatment follow-up, clinical improvement and a decrease in eosinophilic count were observed. Further studies are needed to evaluate the efficacy of treatment, especially in asymptomatic children. PMID- 12724075 TI - [Epidemic outbreak of tuberculosis in a primary and secondary school in Granada (Spain)]. AB - OBJECTIVE: To describe the diagnosis and early treatment of an epidemic outbreak of tuberculosis and determine the utility of polymerase chain reaction (PCR) compared with routine culture in gastric juice. MATERIAL AND METHODS: A computer studies teacher, with clinical features suggestive of tuberculosis and caverns on X-ray, was diagnosed with bacilliferous tuberculosis. Primary health care services carried out a Mantoux test on the school's 387 students as well as on teachers and other staff. The children with a positive Mantoux test underwent laboratory, radiological, and microbiological investigations for one week in the Pediatric Respiratory Unit of Hospital Clinico in Granada. In the teaching and non-teaching staff, active tuberculosis was ruled out through bacilloscopy of sputum samples, Mantoux test, and chest X-ray. RESULTS: In the first screening, the Mantoux test was positive in 67 children. Of these, 7 children were diagnosed with tuberculosis and 60 were found to be infected. Of the 7 children with tuberculosis, five presented positive gastric juice culture in Lowenstein medium while Roche COBAS PCR was negative. In the second screening, 9 children became tuberculin positive. Of these, 8 were diagnosed with tuberculosis and one was infected. Cultures were positive in 3 and PCR was negative. In 77.6 % of the children (59/76), the Mantoux induration was equal to or higher than 18 mm. All of the 15 children with tuberculosis were aged between 9 and 14 years old, except one who was 5 years old. CONCLUSIONS: The Mantoux test remains a basic screening method in diagnosis and epidemiological research, whereas the results of microbiological investigation remain poor and in our study the results DNA were disappointing. The screening of tuberculosis and of other infectious diseases should be more closely monitored in professional groups, such as teachers, that are in contact with large numbers of children. This would identify infected adults and prevent epidemics such as that described in the present study. PMID- 12724076 TI - [Pancreatitis: review of our casuistics in the last 10 years]. AB - OBJECTIVE: Pancreatitis is a rare cause of abdominal pain in childhood. Advances in imaging techniques for pancreatic disease have enabled earlier clinical and etiological diagnosis, thus improving treatment possibilities. The aim of this study was to analyze the etiology, diagnostic methods, and management in patients with pancreatitis diagnosed in our hospital in the last 10 years. MATERIAL AND METHODS: We reviewed the children aged 0-16 years with a diagnosis of pancreatitis admitted to our hospital between 1990 and 2000. Diagnostic criteria were symptoms suggestive of pancreatitis and hyperamylasemia. RESULTS: There were 8 females and 1 male. The median age was 11.5 years (range: 7-16 years). Laboratory findings were as follows: mean amylase level was 1601 U/l and mean lipase level was 506 U/l. Imaging tests: ultrasonography, abdominal computed tomography (CT), and endoscopic retrograde cholangiopancreatography (ERCP) were performed in 1 patient, and magnetic resonance cholangiopancreatography (MRCP) was performed in 2. Etiology was cholelithiasis (2 patients), secondary to anti oncological therapy (3 patients), hypercholesterolemia (1 patient), and idiopathic (3 patients). Medical treatment consisted of diet, analgesia, and octreotide in 1 patient, and antioxidants in 1 patient. Surgery was required in 2 patients. Complications consisted of pancreatic pseudocyst in 2 patients. CONCLUSIONS: The incidence of childhood pancreatitis is low. In our series, mainly teenage girls were affected. The main causes were idiopathic and toxic. Diagnosis was given by elevated amylase and lipase levels in blood and by imaging tests (ultrasonography, abdominal CT, ERCP, MRCP, depending on the patient). Basic treatment consisted of support measures (fasting and analgesia) together with octreotide, antioxidants or surgical treatment, depending on etiology. PMID- 12724077 TI - [Effects of valproate on sexual development]. AB - INTRODUCTION: Valproate use in young girls has been associated with the adverse endocrinological effects of weight gain and hyperandrogenism. Furthermore, polycystic ovaries and hyperinsulinism have been described in adult women. In men and young boys, however, the possible adverse endocrinological effects of valproate have scarcely been analyzed. OBJECTIVES: The aim of this study was to evaluate the effects of valproate treatment on pubertal development, especially the possible hyperandrogenic effects, in girls and boys with epilepsy. MATERIAL AND METHODS: Twenty-three girls and 15 boys (aged 8-16 years old) who were undergoing valproate treatment for epilepsy were compared with 15 control girls and 10 control boys of the same age range. Anthropometric indexes, sexual maturation, and hirsutism scores were evaluated. Serum concentrations of follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone, androstenedione, dehydroepiandrosterone, estradiol, and insulin were measured. Ultrasound examination of ovaries and estimation of bone age through X-ray of the left hand were also performed. RESULTS: Valproate did not affect pubertal development in the study group. No hirsutism or polycystic ovaries were found. Increases in weight, relative weight, and body mass index were observed in the group undergoing valproate treatment, but no statistically significant differences compared with the control group were found. Plasma testosterone was higher in valproate-treated girls (0.71 0.51 ng/ml) than in control girls (0.35 0.15) (p 0.001). This finding was independent of valproate dose and treatment duration. Hyperandrogenism was not found in valproate-treated boys. CONCLUSIONS: Valproate may induce hyperandrogenism in epileptic girls but not in boys. This is an early adverse effect and is independent of the dose used. No changes in normal pubertal development or physical repercussions were found in epileptic patients. PMID- 12724078 TI - [Care of children and adolescents with asthma by primary care physicians: current situation and proposals for improvement]. AB - INTRODUCTION: To provide good asthma care to children and adolescents the following conditions are required: well-trained health professionals with the structure and resources necessary to perform their work efficiently, and client satisfaction as a measure of compliance and self-management. The aims of this study were to determine the current situation of asthma management by primary care pediatricians and to promote improvement of the healthcare and quality of life of these children and their families. MATERIAL AND METHODS: We performed a descriptive study. A questionnaire was completed by primary care pediatricians attending asthma courses given by the Respiratory Team of the Spanish Association of Primary Care Pediatrics in 2001. RESULTS: Three hundred twenty-three questionnaires were completed. The mean age of the pediatricians was 42 years and 70.6 % were women. A total of 7.4 % referred all patients to the pediatric pneumologist and 21.3 % referred all patients to the pediatric allergy clinic; 61.9 % referred only patients with moderate-severe persistent asthma. Concerning items registered in medical records, 32.5 % recorded a codified asthma diagnosis and 61 % included severity classification. Spirometers were available to 48.6 % and peak flow meters to 45.5 %. With regard to lung function measurement, 17.3 % performed spirometry and 33.2 % performed peak expiratory flow. Concerning asthma educational issues, 90.4 % had received education on environmental factors, 81.7 % had received training in the proper use of medication inhalers and 41.2 % had received training in written self-management plans. CONCLUSIONS: This is the first study to present results on the current situation of asthma management by primary care pediatricians in Spain. The results show the need to improve record keeping, facilitate the resources required for diagnosis and follow-up and prioritize education in order to achieve an optimal level of self-management by patients and their families. PMID- 12724079 TI - [Advances in the diagnosis of ENT tumors in childhood]. AB - In the present study we review ENT tumor pathology in childhood. Only the most salient aspects are emphasized and the variety of entities reviewed was restricted. Molecular biology techniques reveal infection by human papilloma virus (types 6 and 11) in 50 % of papillomas, while immunohistochemical techniques are less effective in papilloma virus detection. The myofibroblastic nature of nasal angiofibroma has been demonstrated and its incidence is 25 times more frequent in patients with familial polyposis of the colon. Overexpression of p53 occurs in the initial stages of nasopharyngeal carcinoma, while overexpression of c-myc is correlated with an unfavorable prognosis. Recently, olfactory neuroblastoma has been shown not to express the protein product of the MIC-2 gene (antibody 12E7), thus the hypothesis that it could be a member of the Ewing tumor family (neuroectodermal peripheral tumors) has not been confirmed, although it is a primitive neural tumor. The head and neck rhabdomyosarcoma with the best prognosis is that located in the orbit, and cytogenetic studies have shown chromosomic translocation t(2;13) in 50 % of these childhood tumors when they are of the alveolar-type, while trisomy of chromosome 2 or 20 is more characteristic of the embryonic-type. Currently, any classifying features of ENT lymphomas must be based on the Revised European-American Classification of Lymphoid Neoplasms (REAL). Papillary and medullary carcinomas are the most common histological types of thyroid carcinoma in childhood. Alterations in ret/PTC play a significant role in the pathogenesis of both. PMID- 12724080 TI - [Morbidity and mortality of very-low-birth-weight infants as an indicator of the quality of perinatal care]. AB - OBJECTIVE: To compare the clinical outcomes of a cohort of very low birth weight (VLBW) infants who received healthcare in our unit from 1994-2000 with all the variables included in the Vermont-Oxford Network (VON) database. METHODS: A historical cohort of 417 VLBW live infants born in our center from 1994-2000 was evaluated. The 80 variables of the VON already prospectively included in the unit's database were used and a further 20 variables were added through retrospective review of medical records. The rates of perinatal risk factors, interventions, and causes of morbidity were analyzed and the periods 1994-1997 and 1998-2000 were compared. We also compared these rates with those reported by the VON. RESULTS: Comparison of the results in both periods showed an increase in the percentage of multiple pregnancies and prenatal corticosteroid exposure, as well as in the early use of surfactant and continuous positive pressure. The incidence of intraventricular hemorrhage decreased, but no differences were observed in other outcomes. Our rates of Cesarean sections and multiple births, as well as the use of prenatal steroids, were higher than those of the VON. The outcomes of infants receiving healthcare in our unit were similar to those of the VON but mortality in infants weighing < 800 g was slightly higher, coinciding with higher rates of late sepsis. CONCLUSION: Morbidity rates in VLWB infants receiving care in our unit decreased during the period studied and compared favorably with those reported by the VON. Alltogether, our results indicate that the quality of care in our perinatal center is good. General use of this methodology would permit comparison of outcomes and quality of care across regions and nations, as well as across Europe, in a recently established network (EuroNeoNet.com). PMID- 12724082 TI - [Hepatitis B. The virus, diagnostic techniques, epidemiology, disease and its possible outcomes]. PMID- 12724081 TI - [Retinopathy of prematurity: casuistics between 1996 and 2001]. AB - BACKGROUND: Retinopathy of prematurity (ROP) is a cause of neurosensorial morbidity. OBJECTIVES: To study the incidence, associated risks factors, treatment, and outcome of ROP in premature infants born at less than 32 weeks in our hospital. METHODS: We performed a descriptive study of patients born between the January 1, 1995, and December 31, 2001, in Sant Joan de Deu Hospital in Barcelona (Spain) at <= 32 weeks of gestation who survived until their first month of life. An ocular evaluation was performed between weeks 4 and 6 of life and was repeated every 1-2 weeks until retinal vascularization was complete. Ocular sequelae and visual function were evaluated. Bivariate comparison of groups with and without ROP was performed. RESULTS: Of the 324 patients evaluated, 74 presented ROP (22.8 %), of which 63 patients (21.7 %) were classified as stage 1 or 2 and 11 (3.7 %) as stage 3. An inverse correlation between the incidence of retinopathy and weight and gestational age was found. Threshold disease (3 plus) was detected in 9 patients (16 eyes; 3.1 % of the study sample and 12.1 % of the neonates with retinopathy). All of these neonates were treated with laser therapy. Ocular sequelae were mild in 2.7 % of the patients, moderate in 0.6 % and severe in 0.6 %. The visual function (n 236) of infants with ROP (n 74) was altered in 4 patients (1.7 %). Of these, alterations were severe in 2 patients (0.8 %). Bivariate analysis revealed significant differences (p < 0.001) in low birth weight, gestational age, days of oxygen therapy, days of mechanical ventilation, days of antibiotic therapy, and number of blood transfusions. CONCLUSIONS: In this study the incidence of ROP was similar to that in other centers. Development of ROP was strongly associated with its various risk factors. Severe stages were not seen above 30 weeks of gestational age. The results of laser therapy were optimal, with fewer alterations in ocular examination and visual function than those estimated in patients without treatment. PMID- 12724083 TI - [Hepatitis B. Chronic hepatitis. Outcome and treatment]. PMID- 12724084 TI - [Hepatitis C. The virus, diagnostic methods, epidemiology, self-limiting and chronic disease]. PMID- 12724085 TI - [Hepatitis C. Chronic hepatitis. Outcome and treatment]. PMID- 12724086 TI - [Muscle tumor in a girl with HIV infection: extranodal non-Hodgkin's lymphoma]. AB - Non-Hodgkin's lymphomas are the most common cancers in children with human immunodeficiency virus (HIV) infection. Extranodal disease is the most common form of clinical presentation. Treatment with aggressive chemotherapy and highly active antiretroviral therapy has improved the prognosis of these children. We present a 10-year-old severely immunocompromised girl with HIV infection and non Hodgkin's lymphoma manifested by a muscle mass. PMID- 12724087 TI - [Proteus syndrome: report of two cases]. AB - Proteus syndrome is a complex disorder, consisting of mesodermal and skeletal malformations as well as hamartomatous overgrowth. It is characterized by highly variable presentation and mosaic-pattern lesions. The two cases described herein illustrate the wide polymorphism of this syndrome. The second case also had a pubic fracture, which we have not previously seen reported in the medical literature on Proteus syndrome. PMID- 12724088 TI - [Occult bacteriemia, per se, is not a form of invasive disease]. PMID- 12724089 TI - [Cardiorespiratory arrest related to ophthalmologic examination in premature infants]]. PMID- 12724090 TI - [Early neonatal convulsion: accidental mepivacaine administration]. PMID- 12724091 TI - [Foramina parietalia permagna, an uncommon congenital defect]. PMID- 12724092 TI - [Suppurative lymphadenitis as a complication of BCG vaccination]. PMID- 12724093 TI - [Meningoencephalitis due to Salmonella virchow with unsatisfactory outcome]. PMID- 12724094 TI - [Deformity of the lower left limb and pigmented skin lesions]. PMID- 12724095 TI - Geometrics and dynamics of a rod determine how it is used for reaching. AB - Displacing an object with a hand-held rod provided a simple paradigm for studying tool use. The authors asked how reaching was affected by manipulations of rod properties. Adults held a rod (length =.10 to 1.5 m), with its tip in the air; walked toward an object on a table; chose a place to stop; and displaced the object with the rod's tip. In 3 experiments (Ns = 9, 22, and 17 participants), the authors manipulated rod length, mass, and mass distribution to determine whether and how geometric and dynamic properties affected the chosen distance and the posture. Both the chosen stopping distance and the postures were well accommodated to rod characteristics. Postural adaptations took place only in the arm, which was organized as a synergy. Predictably, rod length explained most of the variance, but small and reliable differences in both distance and posture depended on mass and mass distribution. The chosen distance anticipated not only rod length but also the upcoming posture needed to control the rod. PMID- 12724096 TI - Predictive pointing movements and saccades toward a moving target. AB - The authors investigated whether and, if so, how velocity information is used to control predictive manual pointing movements and saccades. Participants (N = 6) intercepted an occluded moving target as if it were still visible. They kept their eyes fixated while the target moved. The target traveled over a fixed distance and changed its velocity on the way. The presentation time of the final velocity was varied. Both the eye and the hand overshot the slow target and undershot the fast target, particularly when the duration of the final velocity was short. Thus, responses were biased in the direction of the target's initial velocity. The error seemed to arise because participants did not take their latency into account when aiming at the target. Instead, they strategically aimed farther ahead when the target was fast. Amplitude was also more related to the position of velocity change than to final velocity duration. Both findings suggest that target velocity is not extrapolated but that individuals add an increment to the position of velocity change. PMID- 12724097 TI - Effect of practice on effector independence. AB - The authors' primary purpose in the present experiment was to determine if practice changes the extent to which simple motor sequences are effector independent. Contralateral and ipsilateral effector transfers were assessed in 24 participants after 1 (200 trials) and 4 (800 trials) days of practice. The response sequence became increasingly effector dependent; the response structure and the scaling of force on the effector transfer tests were no better after 4 days of practice than after only 1 day, even though retention performance improved substantially. Those results are consistent with the notion that participants refine their movements over extended practice by exploiting the unique characteristics of the effectors. The additional practice results in a more effective movement when the same effectors are used but is of little value when different effectors are required. PMID- 12724098 TI - Development of force adaptation during childhood. AB - Humans learn to make reaching movements in novel dynamic environments by acquiring an internal motor model of their limb dynamics. Here, the authors investigated how 4- to 11-year-old children (N = 39) and adults (N = 7) adapted to changes in arm dynamics, and they examined whether those data support the view that the human brain acquires inverse dynamics models (IDM) during development. While external damping forces were applied, the children learned to perform goal directed forearm flexion movements. After changes in damping, all children showed kinematic aftereffects indicative of a neural controller that still attempted to compensate the no longer existing damping force. With increasing age, the number of trials toward complete adaptation decreased. When damping was present, forearm paths were most perturbed and most variable in the youngest children but were improved in the older children. The findings indicate that the neural representations of limb dynamics are less precise in children and less stable in time than those of adults. Such controller instability might be a primary cause of the high kinematic variability observed in many motor tasks during childhood. Finally, the young children were not able to update those models at the same rate as the older children, who, in turn, adapted more slowly than adults. In conclusion, the ability to adapt to unknown forces is a developmental achievement. The present results are consistent with the view that the acquisition and modification of internal models of the limb dynamics form the basis of that adaptive process. PMID- 12724099 TI - Distraction affects the performance of obstacle avoidance during walking. AB - In this study, dual-task interference in obstacle-avoidance tasks during human walking was examined. Ten healthy young adults participated in the experiment. While they were walking on a treadmill, an obstacle suddenly fell on the treadmill in front of their left leg during either midswing, early stance, or late stance of the ipsilateral leg. Participants were instructed to avoid the obstacle, both as a single task and while they were concurrently performing a cognitive secondary task (dual task). Rates of failure, avoidance strategy, and a number of kinematic parameters were studied under both task conditions. When only a short response time was available, rates of failure on the avoidance task were larger during the dual task than during the single task. Smaller crossing swing velocities were found during the dual task as compared with those observed in the single task. The difference in crossing swing velocities was attributable to increased stiffness of the crossing swing limb. The results of the present study indicated that divided attention affects young and healthy individuals' obstacle avoidance performance during walking. PMID- 12724100 TI - Effects of visual information and task constraints on intersegmental coordination in playground swinging. AB - The authors investigated how and to what extent visual information and associated task constraints are negotiated in the coordinative structure of playground swinging. Participants (N = 20) were invited to pump a swing from rest to a prescribed maximal amplitude under 4 conditions: normal vision, no vision, and 2 visual conditions involving explicit phasing constraints. In the latter conditions, participants were presented with a flow pattern consisting of a periodically expanding and contracting optical structure. They were instructed to phase the swing motion so that the forward turning point coincided with either the maximal size (enhanced optical flow) or the minimal size (reduced optical flow) of the presented flow pattern. Removal of visual information clearly influenced the swinging behavior, in that intersegmental coordination became more stereotyped, reflecting a general stiffening of the swinger. The conditions involving explicit phasing requirements also affected the coordination, but in an opposite way: The coordination became less stereotyped. The two phasing instructions had differential effects: The intersegmental coordination deviated more from normal swinging (i.e., without phasing constraints) when optical flow was enhanced than when it was reduced. Collectively, those findings show that visual information plays a formative role in the coordinative structure of swinging, in that variations of visual information and task constraints were accompanied by subtle yet noticeable changes in intersegmental coordination. PMID- 12724101 TI - Age-related changes in grip force and dynamics of hand movement. AB - The authors investigated whether older adults (n = 16; mean age = 65 years) increased grip force to compensate for load force fluctuations during up and down movements more than young adults did (n = 16; mean age = 24 years) and whether older and young adults exhibited similar adaptation of grip force to alterations in friction associated with changes in object surface texture. As previously reported, older adults used a higher level of grip force than young adults during static holding. Increased grip force was observed in the older group during movement. The increase was appropriate to the lower coefficient of friction estimated for the older group. In both groups, grip force was greater with a smooth than with a rough surface (the latter having the higher coefficient of friction) during static holding and during movement. Moreover, grip force modulation was equally well synchronized with load force fluctuation during movement in the two groups. The authors concluded that changes in organization of grip force with age are well adapted to change in hand-object interface properties. Elevated grip force in older adults does not necessarily signify a fundamental change in synchronizing grip force modulation with load force fluctuation. PMID- 12724102 TI - A methodological note on nonlinear time series analysis: is the open- and closed loop model of Collins and De Luca (1993) a statistical artifact? AB - The authors reexamined, theoretically and empirically, the method proposed by J. J. Collins and C. D. De Luca (1993) for the analysis of center-of-pressure trajectories. The main argument in this article is that Collins and De Luca's approach is not adapted to the analysis of bounded time series and leads to statistical artifacts such as underestimation of the diffusion process for long term intervals. The open- and closed-loop model developed by Collins and De Luca is a direct consequence of those statistical problems. Applying more classical methods, such as rescaled range analysis or detrended fluctuation analysis, the authors show that center-of-pressure trajectories can be modeled as continuous, antipersistent fractional Brownian motion. More specifically, those trajectories behave like 1/f noise, a ubiquitous feature in adaptive biological systems. PMID- 12724104 TI - Research training in mental health and aging: the harvest is plentiful; the laborers, few. PMID- 12724105 TI - It does take a village to care for elderly citizens. PMID- 12724106 TI - Career development and training in geriatric mental health: report of an NIMH workshop. AB - At a National Institute of Mental Health (NIMH)-sponsored meeting, the participants discussed means of increasing the pool of late-life mental-illness researchers. Approaches identified included encouraging retention of junior scientists through greater mentoring and support; creation of research postdoctoral programs by investigators and institutions that lack late-life emphasis; earlier commitment to late-life research with predoctoral training mechanisms; recruitment of ethnic and racial minority scholars into late-life research; and recruitment of newly established researchers through postdoctoral training mechanisms. Federal, public, and private mechanisms need to be better leveraged to grow late-life mental-illness research infrastructure and meet increasing demand and scientific opportunities. PMID- 12724108 TI - Geriatric psychiatry fellowship programs in the United States: findings from the Association of Directors of Geriatric Academic Programs' longitudinal study of training and practice. AB - OBJECTIVE: The authors document the development and growth of geriatric psychiatry fellowship training in the United States (U.S.) through 2002. METHODS: A cross-sectional survey of the 62 U.S. geriatric psychiatry fellowship programs was conducted in Fall 2001. They also analyzed longitudinal data from the American Medical Association (AMA) and the Association of American Medical Colleges' (AAMC) National Graduate Medical Education (GME) Census, along with data from the Accreditation Council for Graduate Medical Education (ACGME). RESULTS: Forty-six (74%) of 62 training directors (TDs) responded. The number of fellowship programs has slowly increased over the past 7 years. During 2001-2002, a total of 94 fellows were in training (all years of training). Seventy-eight percent (N=36) of responding programs offered only 1-year fellowship training experiences. TDs reported that application rates for fellowship positions were stable during the academic years 1999-2002, with a median number of eight applications per program for first-year positions in 2001-2002. The fill-rate for first-year geriatric psychiatry fellowship positions dropped from 84% in 1999 2000 to 61% in 2001-2002. During 2001-2002, 73% of programs reported having two or fewer first-year fellows, and 16% had no first-year fellows. Seventeen programs reported having no U.S. medical school graduates (USMGs) as first-year fellows. CONCLUSION: Recruiting high-quality USMGs into geriatric psychiatry fellowship programs remains a challenge. Furthermore, retaining first-year fellows for additional years of academic training has been difficult. Findings indicate that specific strategies need to be developed to stimulate undergraduate and graduate interest in careers in clinical and academic geriatric psychiatry. PMID- 12724107 TI - Evolution of geriatric medicine fellowship training in the United States. AB - The authors describe the history of subspecialty training in geriatric medicine and geriatric psychiatry, from the time before the formal accreditation of geriatric medicine fellowship programs, in 1988, through the following decade of 2-year accreditation programs, from 1988 to 1997, and then the recent experience with 1-year accredited geriatric medicine programs, from 1998 to the present. They present the history and development of the accrediting organizations, the development of relevant tests and curricula, and the opportunities for grants, funding, and career development, as well as a summary of present status and satisfaction of current practitioners, and provide recommendations for enhancing recruitment and retention. PMID- 12724109 TI - Effect of training and other influences on the development of career interest in geriatric psychiatry. AB - OBJECTIVE: The authors examined the relationship between the timing and nature of educational exposure to geriatric psychiatry and other potential influences and subsequent development of career interest in geriatric psychiatry. METHODS: A 46 item survey was distributed to residents and fellows who attended one of the two sponsored programs for residents at the three American Association of Geriatric Psychiatry (AAGP) annual meetings held between 2000 and 2002, inclusive. RESULTS: Ninety-three percent of attendees responded (N=184). Sixty-five percent first developed interest in geriatric psychiatry during their residency years (the majority during their PGY1 or PGY2 year). The timing of individualized teaching exposure, as well as lectures in geriatric psychiatry, was associated with the development of first interest in the field. The most important influences on the development of interest in the field included specific teacher attributes, training experiences, personal experiences with seniors, and characteristics cited as unique to geriatric psychiatry, such as the medical, neuropsychiatric, and multifactorial nature of the field. Patient personal histories and outcomes, as well as non-educational experiences with seniors and cultural attitudes, also contributed to interest. CONCLUSION: It behooves geriatric psychiatry programs to create exemplary educators and commit them to teaching in the early years of general psychiatry programs as well as in medical school. These educators should be identifying potential recruits by enquiring about trainees' previous experiences with older persons as well as emphasizing the unique aspects of geriatric psychiatry that are attractive to trainees. PMID- 12724110 TI - The Stressful Caregiving Adult Reactions to Experiences of Dying (SCARED) Scale: a measure for assessing caregiver exposure to distress in terminal care. AB - OBJECTIVE: The authors evaluated the performance of the Stressful Caregiving Adult Reactions to Experiences of Dying (SCARED) scale, a new tool to assess caregiver exposure to patient distress, and the fear and helplessness evoked by these experiences. METHODS: The SCARED was administered to 76 hospice-patient caregivers (e.g., family members). Major depressive disorder (MDD) was diagnosed with the Structured Clinical Interview for the DSM-IV; complicated grief (CG) "caseness" was diagnosed with the Inventory of Complicated Grief-Caregiver items, and quality-of-life domains were assessed with the Medical Outcomes Survey Short Form-36. RESULTS: Respondents endorsed frequent exposure to each SCARED experience-from 30.3% who mistakenly thought the patient had died, to 80.3% who witnessed the patient in severe pain. Adjusted analyses revealed that the odds of MDD increased by 3.08 for each standard-deviation increase in the SCARED event frequency score and that higher SCARED scores were positively associated (p <0.05) with social and role impairment, less energy, and more negative health perceptions. CONCLUSION: The SCARED may be a clinically useful tool for identifying caregivers at risk for MDD and quality-of-life impairments, as well as potential aspects of caregiving to target for treatment. PMID- 12724112 TI - Comorbid medical conditions among depressed elderly patients discharged home after acute psychiatric care. AB - OBJECTIVE: The authors examined physical illness among elderly patients hospitalized for treatment of major depression, the impact of comorbidity on functional status, and the burden of comorbidity on post-discharge service needs. METHODS: Data for this cross-sectional study were derived from patient interviews and abstracted from hospital charts. The sample comprised 195 older adults hospitalized for treatment of depression on the geropsychiatric unit of a large urban teaching hospital and discharged to home. Medical comorbidity was measured with the Cumulative Illness Rating Scale for Geriatrics. A psychiatrist confirmed DSM-IV for Axis I diagnosis of depression, and the Geriatric Depression Scale measured depression severity. Unit nurses administered the Mini-Mental State Exam. They assessed functional dependency via the OARS Multidimensional Functional Assessment Questionnaire. Service needs post-discharge were measured from physician discharge orders and patient scores on standardized tests. RESULTS: Almost three-fourths of depressed elderly patients had at least one comorbid condition requiring first-line treatment; nearly half had two, and one fourth had three or more. Comorbid physical illness and cognitive impairment was significantly and negatively associated with elderly patients' functional impairment at discharge. Depressed patients with higher medical comorbidity had significantly more needs for services after they left acute care. CONCLUSION: Medical comorbidity needs to be assessed and considered in planning for post acute care for depressed elderly patients discharged home. PMID- 12724111 TI - Treatment of poststroke generalized anxiety disorder comorbid with poststroke depression: merged analysis of nortriptyline trials. AB - OBJECTIVE: The existence of anxiety disorders plays an important role in the prognosis and associated impairment among patients with poststroke depression. The authors examined the efficacy of nortriptyline treatment for patients with comorbid generalized anxiety disorder (GAD) and depression after stroke. METHODS: Data from three studies were merged to provide 27 patients with comorbid GAD and depression, who participated in double-blind treatment studies comparing nortriptyline (N=13) and placebo (N=14). Severity of anxiety was measured with the Hamilton Rating Scale for Anxiety (Ham-A), and severity of depression was measured with the Hamilton Rating Scale for Depression (Ham-D). Activities of daily living were assessed by use of the Johns Hopkins Functioning Inventory (JHFI). RESULTS: There were no significant differences between the nortriptyline and placebo groups in demographic characteristics, stroke type, and neurological findings. Patients receiving nortriptyline treatment showed significantly greater improvement on the Ham-A, Ham-D, and JHFI than patients receiving placebo. The anxiety symptoms showed earlier improvement than depressive symptoms in patients treated with nortriptyline. CONCLUSIONS: These findings suggest that poststroke GAD comorbid with poststroke depression may be effectively treated with nortriptyline, and data indicate the need for a trial specifically designed to examine treatment of anxiety disorder. PMID- 12724114 TI - Subspecialty certification in geriatric psychiatry. AB - OBJECTIVE: The authors describe the development of certification and recertification processes for geriatric psychiatry, the American Board of Psychiatry and Neurology's second subspecialty in psychiatry. METHODS: Authors referenced databases of the certifying organizations and consulted published surveys of examinees. RESULTS: Since the first certification examination was administered in 1991, 2,595 certificates have been awarded. Because the certificates are time-limited, diplomates must sit for recertification every 10 years. Thus far, about 63% of the diplomates who have needed to do so have been recertified. CONCLUSION: It is estimated that 5,000 geriatric psychiatrists will be needed to meet future healthcare needs. Thus, the demand for this expertise will be increasing, and it is anticipated that board-certified geriatric psychiatrists will continue to play leading roles in educating practitioners from multiple disciplines. PMID- 12724113 TI - Partial response as a predictor of outcome in geriatric depression. AB - OBJECTIVE: Although geriatric depression generally responds to treatment, patients are often left with residual symptoms after acute treatment. Data are lacking on the eventual outcomes of individuals who are partial responders or poor responders to initial treatment. METHODS: The authors examined 200 patients initially treated with a treatment-based guideline approach that allowed for physician choice. RESULTS: After 3 months, 34.0% of patients were remitted, 37.5% had achieved a partial response, and 28.5% were still highly symptomatic. After 6 months of treatment, 51.1% of patients were remitted, 36.5% were partial responders, and 12.5% were still experiencing significant depressive symptoms. Cumulative remission rates over 18 months of treatment were significantly higher among partial responders than among highly symptomatic patients at 6 months, but not at 3 months. In terms of relapse, defined as a Montgomery-Asberg Depression Rating Scale score greater than 15, 42.0% of the remitted and partial-responder groups had relapsed by 18 months. When compared with remitted patients, partial responders at 6 months were more likely to experience a relapse. There were no differences in relapse rate between the two groups at 3 months. CONCLUSION: These results suggest that during ongoing active treatment of geriatric depression, the level of improvement at 6 months, but not 3 months, is predictive of subsequent course. PMID- 12724115 TI - Challenges to the transition to independent investigator in geriatric mental health. AB - OBJECTIVE: The author reports on the extent to which junior investigators in geriatric mental health successfully make the transition to independent scientists. METHODS: The NIH CRISP database identified all NIMH Level-1 career awards (K01/K07/K08/K23/ R29) in geriatric mental health completed from 1992 to 2001. RESULTS: Of 46 awardees, 14 (30.4%) achieved R01 funding within 1 year of completion. An additional six investigators subsequently achieved R01s in the remaining follow-up period (ranging from 2 to 10 years), for a total of 43.5% (20/46). CONCLUSIONS: These data underscore the relatively small number of junior investigators who successfully make the transition to independent scientist. Their numbers underscore the importance of efforts by the field of geriatric mental health aimed at both increasing the flow into this career as well as ensuring the retention and success of junior investigators at the critical transition to independent investigators. PMID- 12724116 TI - Alcohol dependence among elderly French inpatients. AB - OBJECTIVE/METHODS: The authors compared groups of heavier and lighter drinkers in a geriatric hospital inpatient unit in France, using questionnaire measures of alcohol dependence, sociodemographic characteristics, and cognitive status. RESULTS: Alcohol consumption is more frequent in men than in women; 9% of the geriatric patients (3% of women and 18% of men) show symptoms of alcohol dependence. Alcohol dependence is also associated with benzodiazepine treatment and nicotine dependence. CONCLUSION: Elderly male patients hospitalized in the geriatric unit and receiving benzodiazepines should thus be identified as a population at risk for alcohol dependence, and medical practitioners should be alert to the possibility of overprescribing benzodiazepines past the acute withdrawal stage. PMID- 12724117 TI - Age and major depression after mild traumatic brain injury. AB - OBJECTIVE: The authors investigated the relationship between age and major depression in the acute period following mild traumatic brain injury (TBI). METHODS: Patients with mild TBI (N=210) were assessed for the presence of major depression with the Structured Clinical Interview for DSM-IV. RESULTS: Older patients (age 60-plus) had lower rates of major depression than younger patients. CONCLUSION: Older patients seem to be relatively resilient to major depression shortly after mild TBI. PMID- 12724118 TI - Cognitive burden and excess Lewy-body pathology in the Lewy-body variant of Alzheimer disease. AB - OBJECTIVES: Authors compared the degrees of cognitive deficit among individuals with Alzheimer disease (AD), the Lewy-body variant of AD (LBV), and "pure" dementia with Lewy bodies (DLB); and compared cortical Lewy body (LB) counts in LBV versus DLB and neuritic plaque and neurofibrillary tangle severity in LBV versus AD. METHODS: Authors examined brain specimens from consecutive autopsies of elderly nursing home subjects. Numbers and densities of plaques, Lewy bodies, and tangle severity were determined in multiple cortical regions, and demographic and clinical variables were compared among the three groups. RESULTS: The three groups did not differ in demographic or clinical variables. The LBV group was significantly more impaired than the other groups. Cortical LB counts were significantly higher in LBV than in DLB. There was no evidence of increased plaque or tangle severity in LBV than in AD. CONCLUSION: The co-occurrence of AD and LB pathology is associated with higher numbers of LBs and more severe dementia than when classical AD or LB lesions occur alone. PMID- 12724120 TI - Commentary on letter: atypical antipsychotics and coma. PMID- 12724121 TI - Chlorine-induced injury to the airways in mice. AB - Exposure to chlorine gas (Cl2) causes occupational asthma that we hypothesized occurs through the induction of airway inflammation and airway hyperresponsiveness by oxidative damage. Respiratory mechanics and airway responsiveness to methacholine were assessed in A/J mice 24 hours after a 5 minute exposure to 100, 200, 400, or 800 ppm Cl2 and 2 and 7 days after inhalation of 400 ppm Cl2. Airway responsiveness was higher 24 hours after 400 and 800 ppm Cl2. Responsiveness after inhalation of 400 ppm Cl2 returned to normal by 2 days but was again elevated at 7 days. Airway epithelial loss, patchy alveolar damage, proteinaceous exudates, and inflammatory cells within alveolar walls were observed in animals exposed to 800 ppm Cl2. Macrophages, granulocytes, epithelial cells, and nitrate/nitrite levels increased in lung lavage fluid. Increased inducible nitric oxide synthase expression and oxidation of lung proteins were observed. Epithelial cells and alveolar macrophages from mice exposed to 800 ppm Cl2 stained for 3-nitrotyrosine residues. Inhibition of inducible nitric oxide synthase with 1400W (1 mg/kg) abrogated the Cl2-induced changes in responsiveness. We conclude that chlorine exposure causes functional and pathological changes in the airways associated with oxidative stress. Inducible nitric oxide synthase is involved in the induction of changes in responsiveness to methacholine. PMID- 12724122 TI - Fungal levels in the home and lower respiratory tract illnesses in the first year of life. AB - The association between home dampness and lower respiratory symptoms in children has been well documented. Whether fungal exposures contribute to this association is uncertain. In a prospective birth cohort of 499 children of parents with asthma/allergies, we examined in-home fungal concentrations as predictors of lower respiratory illnesses (LRI) (croup, pneumonia, bronchitis, and bronchiolitis) in the first year. In multivariate analyses, we found a significant increased relative risk (RR) between LRI and high levels (more than the 90th percentile) of airborne Penicillium (RR = 1.73, 95% confidence interval [CI], 1.23, 2.43), dust-borne Cladosporium (RR = 1.52; CI, 1.02, 2.25), Zygomycetes (RR = 1.96; CI, 1.35, 2.83), and Alternaria (RR = 1.51; CI, 1.00, 2.28), after controlling for sex, presence of water damage or visible mold/mildew, born in winter, breastfeeding, and being exposed to other children through siblings. In a multivariate analysis, the RR of LRI was elevated in households with any fungal level at more than the 90th percentile (RR = 1.86; CI, 1.21, 2.88). Exposure to high fungal levels increased the risk of LRI in infancy, even for infants with nonwheezing LRI. Actual mechanisms remain unknown, but fungi and their components (glucans, mycotoxins, and proteins) may increase the risk of LRI by acting as irritants or through increasing susceptibility to infection. PMID- 12724123 TI - beta-Lapachone reduces endotoxin-induced macrophage activation and lung edema and mortality. AB - beta-Lapachone, a 1,2-naphthoquinone, is a novel chemotherapeutic agent. It has been shown to be capable of suppressing inducible nitric oxide synthase expression and function in rat alveolar macrophages. The authors further performed experiments to examine the molecular mechanism of beta-lapachone on LPS induced responses in rat alveolar macrophages and to evaluate its in vivo antiinflammatory effect. A significant increase in nitrite production and inducible nitric oxide synthase expression was elicited in macrophages treated with LPS that was inhibited by coincubation with beta-lapachone. beta-Lapachone could also inhibit the production of tumor necrosis factor-alpha induced by LPS. LPS induces protein tyrosine phosphorylation and nuclear factor-kappaB binding activity by gel mobility shift assay in macrophages. These events were significantly inhibited by beta-lapachone. Furthermore, beta-lapachone in vivo protected against the induction of lung edema, lung-inducible nitric oxide synthase protein expression and nuclear factor-kappaB activation, lethality, and increased plasma nitrite and serum tumor necrosis factor-alpha levels induced by LPS. These results indicate that beta-lapachone suppresses inducible nitric oxide synthase induction and tumor necrosis factor-alpha production mediated by the inhibition of protein tyrosine phosphorylation and nuclear factor-kappaB activation caused by LPS. This results in a beneficial effect in an animal model of sepsis. PMID- 12724124 TI - Phenotypic and functional characterization of blood gammadelta T cells in sleep apnea. AB - Hypoxia-induced lymphocyte dysfunction may be implicated in endothelial cell damage in obstructive sleep apnea (OSA) syndrome. gammadelta T cells' unique migration, cytotoxic features, and accumulation in atherosclerotic plaques are considered critical in cardiovascular disorders. We characterized the phenotype, cytokine profile, adhesion properties, and cytotoxicity of gammadelta T cells in patients with OSA and control subjects. The following is a summary of our major findings regarding OSA gammadelta T cells: (1) a significant increase in the expression of the inhibitory natural killer B1 receptors was found; (2) the intracellular content of proinflammatory cytokines tumor necrosis factor (TNF) alpha and interleukin-8 was increased, and the content of the antiinflammatory cytokine interleukin-10 was decreased; (3) gammadelta T cells of patients with OSA adhered significantly more avidly to nonactivated endothelial cells in culture than those of control subjects; (4) L-selectin expression was higher; (5) anti-E/P-selectin antibodies and anti-TNF-alpha antibodies decreased the adhesion index of OSA gammadelta T lymphocytes/endothelial cells but not of control subjects; and (6) cytotoxicity of OSA gammadelta T lymphocytes against endothelial cells in culture was 2.5-fold higher than that of control subjects and could be prevented by pretreatment with anti-TNF-alpha. Collectively these data implicate gammadelta T lymphocyte function in atherogenic sequelae in OSA. PMID- 12724125 TI - Effect of oral appliance therapy on upper airway collapsibility in obstructive sleep apnea. AB - Oral appliance therapy is emerging as an alternative to continuous positive airway pressure for the treatment of obstructive sleep apnea (OSA). However, its precise mechanisms of action are yet to be defined. We examined the effect of a mandibular advancement splint (MAS) on upper airway collapsibility during sleep in OSA. Ten patients with proven OSA had a custom-made MAS incrementally adjusted during an acclimatization period until the maximum comfortable limit of mandibular advancement was reached. Polysomnography with the splint was then performed. After a 1-week washout period, upper airway closing pressures during sleep (with and without MAS) were determined. Significant improvements with MAS therapy were seen in the apnea/hypopnea index (25.0 +/- 3.1 vs. 13.2 +/- 4.5/hour, p < 0.03) and upper airway closing pressure in Stage 2 sleep (-1.6 +/- 0.4 vs. -3.9 +/- 0.6 cm H2O, p < 0.01) and in slow wave sleep (-2.5 +/- 0.7 vs. 4.7 +/- 0.6 cm H2O, p < 0.02) compared with no therapy. These preliminary data indicate that MAS therapy is associated with improved upper airway collapsibility during sleep. The mediators of this effect remain to be determined. PMID- 12724126 TI - Effect of diesel on chemokines and chemokine receptors involved in helper T cell type 1/type 2 recruitment in patients with asthma. AB - The objective of this study was to evaluate if diesel exhausts could favor helper T cell type (Th) 2-associated allergic reactions either through an increased production of Th2-associated chemokines and of their associated receptors or through a decrease of Th1-attracting chemokines and chemokine receptors. Diesel but not allergen exposure of peripheral blood mononuclear cells from subjects with allergy induced a release of I-309, whereas both diesel and Der p 1 induced an early but transient release of monokine induced by IFN-gamma and a late release of pulmonary and activation-regulated chemokine. Although both Th1- and Th2-attracting chemokines were induced, the resulting effect was an increased chemotactic activity on Th2 but not Th1 cells. Surprisingly, diesel induced a late increase in the expression of the Th1-associated CXC receptor 3 and CC receptor 5. T cell CXC receptor 3 upregulation was not associated with an increased migration to its ligands. These two antagonistic effects have been previously reported as a scavenger mechanism to clear chemokines. Altogether, these results suggest that diesel, even without allergen, may amplify a type 2 immune response but that it can also increase late Th1-associated chemokine receptor expression, perhaps as a scavenger mechanism to clear pro-Th1 chemokines and promote the Th2 pathway. PMID- 12724127 TI - Human pulmonary chimerism after hematopoietic stem cell transplantation. AB - Many of the body's tissues once thought to be only locally regenerative may, in fact, be actively replaced by circulating stem cells after hematopoietic stem cell transplantation. Localization of donor-derived cells ("chimerism") has recently been shown to occur in the lungs of mice after either hematopoietic stem cell transplantation or infusion of cultured marrow. To determine whether tissues of the human lung might be similarly derived from extrapulmonary sources, we examined lung specimens from a retrospective cohort of female allogeneic hematopoietic stem cell transplant recipients who received stem cells from male donors. Tissue samples from three such patients who had undergone diagnostic lung biopsy or autopsy were examined. Slides were stained by immunohistochemistry for cytokeratin (epithelium) and platelet endothelial cell adhesion molecule, CD31 (PECAM) (endothelium) and were imaged and then examined by fluorescent in situ hybridization analysis to identify male cells. The resulting overlapping in situ hybridization and immunohistochemistry images were examined for the presence and, if present, cell type of donor cells in the lung. We found significant rates of epithelial (2.5-8.0%) and endothelial (37.5-42.3%) chimerism. These results suggest that significant chimerism of the human lung may follow hematopoietic stem cell transplantation and that adult human stem cells could potentially play a therapeutic role in treatment of the damaged lung. PMID- 12724128 TI - Regulatory volume increase is associated with p38 kinase-dependent actin cytoskeleton remodeling in rat kidney MTAL. AB - The kidney medulla is physiologically exposed to variations in extracellular osmolality. In response to hypertonic cell shrinkage, cells of the rat kidney medullary thick ascending limb of Henle's loop undergo p38 kinase-dependent regulatory volume increase (RVI). In the present study, we investigated the role of actin cytoskeleton reorganization in this process. Addition of hyperosmotic NaCl or sucrose, which activates MAP kinases and reduces cellular volume, induced a sustained actin polymerization occurring after 10 min and concurrently with RVI. In contrast, hyperosmotic urea, which does not modify MAP kinase activity and cellular volume, did not induce sustained actin polymerization. Fluorescence microscopy revealed that hyperosmotic NaCl and sucrose, but not urea, induced the redistribution of F-actin from a dense cortical ring to a diffuse network of actin bundles. Stabilization of actin filaments by jasplakinolide and inhibition of the generation of new actin filaments by swinholide A prevented RVI, whereas depolymerization of actin filaments by latrunculin B attenuated cell shrinkage and enhanced RVI. These actin-interfering drugs did not alter extracellular regulated kinase and p38 kinase activation under hypertonic conditions. Similar to swinholide A, inhibiting p38 kinase with SB-203580 abolished sustained actin polymerization, actin redistribution, and decreased RVI efficacy. We therefore propose that in rat kidney the medullary thick ascending limb of Henle's loop exposed to extracellular hypertonicity, p38 kinase activation induces depolymerization of the F-actin cortical ring and polymerization of a dense diffuse F-actin network that both contribute to increase RVI efficacy. PMID- 12724129 TI - Characterization of Ca2+-activated Cl- currents in mouse kidney inner medullary collecting duct cells. AB - Ca2+-activated Cl- (ClCa) channels were characterized biophysically and pharmacologically in a mouse kidney inner medullary collecting duct cell line, IMCD-K2. Whole cell recording was performed with symmetrical N-methyl-d-glucamine chloride (NMDG)-Cl in the intracellular and extracellular solutions, and the intracellular Ca2+ concentration ([Ca2+]i) was adjusted with Ca2+-EGTA buffers. The amplitude of the current was dependent on [Ca2+]i. [Ca2+]i <800 nM strongly activated outwardly rectifying Cl- currents, whereas high Ca2+ (21 microM) elicited time-independent currents that did not rectify. The currents activated at low [Ca2+] exhibited time-dependent activation and deactivation. The affinity of the channel for Ca2+ was voltage dependent. The EC50 for Ca2+ was approximately 0.4 microM at +100 mV and approximately 1.0 microM at -100 mV. The Cl- channel blocker niflumic acid in the bath equally inhibited both inward and outward currents reversibly, with a Ki = 7.6 microM. DIDS, diphenylamine-2 carboxylic acid, and anthracene-9-carboxylic acid reversibly inhibited outward currents in a voltage-dependent manner. DTT slowly inhibited the currents, but tamoxifen did not. Comparing the biophysical and pharmacological properties, we conclude that IMCD-K2 cells express the same type of ClCa channels as those we have described in detail in Xenopus laevis oocytes (Qu Z and Hartzell HC. J Biol Chem 276: 18423-18429, 2001). PMID- 12724130 TI - Renal uptake of myoglobin is mediated by the endocytic receptors megalin and cubilin. AB - Nephrotoxicity of myoglobin is well recognized as playing a part in the development of acute renal failure in settings of myoglobinuria. However, the molecular mechanism of myoglobin uptake in renal proximal tubules has not been clarified. Here, we report that the endocytic receptors megalin and cubilin are involved in renal reabsorption of myoglobin. Both receptors were captured from solubilized renal brush-border membranes by affinity chromatography using myoglobin-Sepharose. Myoglobin bound to purified megalin and cubilin with Kd values of 2.0 and 3 microM, respectively, as evaluated by surface plasmon resonance analysis. Apomyoglobin bound to megalin with the same affinity, and the affinity of apomyoglobin to cubilin was reduced (Kd = 5 microM). Radioiodinated myoglobin could be displaced by apomyoglobin in inhibition studies using isolated renal brush-border membranes (Ki approximately 2 microM). Receptor-associated protein as well as antibodies directed against megalin and cubilin markedly inhibited the uptake of fluorescent-labeled myoglobin by cultured yolk sac BN-16 cells. The significance of megalin- and cubilin-mediated endocytosis for myoglobin uptake in vivo was demonstrated by use of kidney-specific megalin knockout mice. Injected myoglobin was extensively reabsorbed by megalin expressing proximal tubular cells, whereas there was very little uptake in the megalin-deficient cells. In conclusion, this study establishes the molecular mechanism of myoglobin uptake in the renal proximal tubule involving the endocytic receptors megalin and cubilin. Identification of the receptors for tubular uptake of myoglobin may be essential for development of new therapeutic strategies for myoglobinuric acute renal failure. PMID- 12724131 TI - A 5-HT4 agonist mosapride enhances rectorectal and rectoanal reflexes in guinea pigs. AB - The rectal distension-evoked reflex rectal (R-R) contractions and internal anal sphincter (R-IAS) relaxations in guinea pigs were generated through the extrinsic sacral excitatory nerve pathway (pelvic nerves) and the intrinsic cholinergic excitatory and nitrergic inhibitory nerve pathways. The aim of the present study was to evaluate whether a prokinetic benzamide, mosapride, enhances the R-R and R IAS reflexes mediated via 5-HT4 receptors in the guinea pig. The mechanical activities of the R and IAS were recorded with a balloon connected to a pressure transducer and a strain gauge force transducer in the anesthetized guinea pig with intact spinal-intestinal pathways. Gradual and sustained rectal distension evoked R-R contractions and synchronous R-IAS relaxations. Mosapride (0.1-1.0 mg/kg i.v.) dose-dependently enhanced both R-R and R-IAS reflex responses. Reflex indexes for R-R and R-IAS maximally increased from 1.0 (control) to 1.92 and 1.88, respectively. A specific 5-HT4 receptor antagonist, GR 113808 (1.0 mg/kg i.v.), antagonized the enhancement of the R-R and R-IAS reflexes induced by mosapride 1.0 mg/kg i.v. The present results indicate that mosapride enhanced the R-R and R-IAS reflexes mediated through 5-HT4 receptors. PMID- 12724133 TI - New method to study oxidative damage and antioxidants in the human small bowel: effects of iron application. AB - Iron may induce oxidative damage to the intestinal mucosa by its catalyzing role in the formation of highly reactive hydroxyl radicals. This study aimed to determine iron-induced oxidative damage provoked by a single clinical dosage of ferrous sulfate and to elucidate the antioxidant defense mechanisms in the human small intestine in vivo. A double-lumen perfusion tube was positioned orogastrically into a 40-cm segment of the proximal small intestine in six healthy volunteers (25 +/- 5 yr). The segment was perfused with saline and subsequently with saline containing 80 mg iron as ferrous sulfate at a rate of 10 ml/min. Intestinal fluid samples were collected at 15-min intervals. Thiobarbituric acid reactive substances concentrations as an indicator of lipid peroxidation increased significantly from 0.07 microM (range, 0-0.33 microM) during saline perfusion to 3.35 microM (range, 1.19-7.27 microM) during iron perfusion (P < 0.05). Nonprotein antioxidant capacity increased significantly from 474 microM (range, 162-748 microM) to 1,314 microM (range, 674-1,542 microM) (P < 0.05). These data show that a single dosage of ferrous sulfate induces oxidative damage and the subsequent release of an antioxidant in the small intestine in vivo in healthy volunteers. PMID- 12724132 TI - Enterocyte apoptosis after enterectomy in mice is activated independent of the extrinsic death receptor pathway. AB - Intestinal adaptation following small bowel resection (SBR) is associated with greater rates of enterocyte apoptosis by unknown mechanism(s). Because postresection adaptation is associated with increased translocation of luminal bacteria, we sought to characterize the role for the extrinsic, death receptor pathway for the activation of enterocyte apoptosis after massive SBR. We first performed SBR or sham operations in mice, and the temporal expression of caspases 8, 9, and 3, death receptors tumor necrosis factor receptor-1 (TNFR1) and Fas and corresponding ligands (TNF and Fas ligand) was determined in the remnant intestine at various postoperative time points. Ileal TNFR1 and Fas expression were then measured after SBR in the setting of increased (waved-2 mice) or decreased (exogenous EGF administration) apoptosis. Finally, intestinal adaptation and apoptosis were recorded in the remnant ileum after SBR in TNFR1 null and Fas-null mice. The expression of death receptor family proteins and caspases demonstrated only modest changes after SBR and did not correlate with the histological appearance of apoptosis. In the setting of accelerated apoptosis, TNFR1 and Fas expression were paradoxically decreased. Apoptotic and adaptive responses were preserved in both TNFR1-null and Fas-null mice. These results suggest that the mechanism for increased enterocyte apoptosis following massive SBR does not appear to involve the extrinsic, death receptor-mediated pathway. PMID- 12724134 TI - Amelioration of TNBS-induced colon inflammation in rats by phospholipase A2 inhibitor. AB - The pathophysiology of inflammatory bowel disease (IBD) involves the production of diverse lipid mediators, namely eicosanoids, lysophospholipids, and platelet activating factor, in which phospholipase A2 (PLA2) is the key enzyme. Accordingly, it has been postulated that control of lipid mediator production by inhibition of PLA2 would be useful for the treatment of IBD. This hypothesis was tested in the present study by examining the therapeutic effect of a novel extracellular PLA2 inhibitor (ExPLI), composed of carboxymethylcellulose-linked phosphatidylethanolamine (CMPE), on trinitrobenzenesulfonic acid-induced colitis. Intraperitoneal administration of CMPE suppressed the colitis as measured by mortality rate, intestinal permeability, plasma PLA2 activity, intestinal myeloperoxidase activity, and histological morphometry. Current therapeutic approaches for inflammatory conditions focus on the selective control of a lipid mediator(s) (e.g., prostaglandins or leukotrienes). The present study supports the concept that inclusive control of lipid mediator production by PLA2 inhibition is a plausible approach to the treatment of colitis and introduces the ExPLIs as a prototype of a novel NSAID for the treatment of intestinal inflammation. PMID- 12724135 TI - Ca2+ sparks are initiated by Ca2+ entry in embryonic mouse skeletal muscle and decrease in frequency postnatally. AB - "Spontaneous" Ca2+ sparks and ryanodine receptor type 3 (RyR3) expression are readily detected in embryonic mammalian skeletal muscle but not in adult mammalian muscle, which rarely exhibits Ca2+ sparks and expresses predominantly RyR1. We have used confocal fluorescence imaging and systematic sampling of enzymatically dissociated single striated muscle fibers containing the Ca2+ indicator dye fluo 4 to show that the frequency of spontaneous Ca2+ sparks decreases dramatically from embryonic day 18 (E18) to postnatal day 14 (P14) in mouse diaphragm and from P1 to P14 in mouse extensor digitorum longus fibers. In contrast, the relative levels of RyR3 to RyR1 protein remained constant in diaphragm muscles from E18 to P14, indicating that changes in relative levels of RyR isoform expression did not cause the decline in Ca2+ spark frequency. E18 diaphragm fibers were used to investigate possible mechanisms underlying spark initiation in embryonic fibers. Spark frequency increased or decreased, respectively, when E18 diaphragm fibers were exposed to 8 or 0 mM Ca2+ in the extracellular Ringer solution, with no change in either the average resting fiber fluo 4 fluorescence or the average properties of the sparks. Either CoCl2 (5 mM) or nifedipine (30 microM) markedly decreased spark frequency in E18 diaphragm fibers. These results indicate that Ca2+ sparks may be triggered by locally elevated [Ca2+] due to Ca2+ influx via dihydropyridine receptor L-type Ca2+ channels in embryonic mammalian skeletal muscle. PMID- 12724136 TI - Myosin II is present in gastric parietal cells and required for lamellipodial dynamics associated with cell activation. AB - Nonmuscle myosin II has been shown to participate in organizing the actin cytoskeleton in polarized epithelial cells. Vectorial acid secretion in cultured parietal cells involves translocation of proton pumps from cytoplasmic vesicular membranes to the apical plasma membrane vacuole with coordinated lamellipodial dynamics at the basolateral membrane. Here we identify nonmuscle myosin II in rabbit gastric parietal cells. Western blots with isoform-specific antibodies indicate that myosin IIA is present in both cytosolic and particulate membrane fractions whereas the IIB isoform is associated only with particulate fractions. Immunofluorescent staining demonstrates that myosin IIA is diffusely located throughout the cytoplasm of resting parietal cells. However, after stimulation, myosin IIA is rapidly redistributed to lamellipodial extensions at the cell periphery; virtually all the cytoplasmic myosin IIA joins the newly formed basolateral membrane extensions. 2,3-Butanedione monoximine (BDM), a myosin ATPase inhibitor, greatly diminishes the lamellipodial dynamics elicited by stimulation and retains the pattern of myosin IIA cytoplasmic staining. However, BDM had no apparent effect on the stimulation associated redistribution of H,K ATPase from a cytoplasmic membrane compartment to apical membrane vacuoles. The myosin light chain kinase inhibitor 1-(5-iodonaphthalene-1-sulfonyl)-1H-hexahydro 1,4-diazepine (ML-7) also did not alter the stimulation-associated recruitment of H,K-ATPase to apical membrane vacuoles, but unlike BDM it had relatively minor inhibitory effects on lamellipodial dynamics. We conclude that specific disruption of the basolateral actomyosin cytoskeleton has no demonstrable effect on recruitment of H,K-ATPase-rich vesicles into the apical secretory membrane. However, myosin II plays an important role in regulating lamellipodial dynamics and cortical actomyosin associated with parietal cell activation. PMID- 12724137 TI - Depolarization-induced contraction and SR function in mechanically skinned muscle fibers from dystrophic mdx mice. AB - Dystrophin is absent in muscle fibers of patients with Duchenne muscular dystrophy (DMD) and in muscle fibers from the mdx mouse, an animal model of DMD. Disrupted excitation-contraction (E-C) coupling has been postulated to be a functional consequence of the lack of dystrophin, although the evidence for this is not entirely clear. We used mechanically skinned fibers (with a sealed transverse tubular system) prepared from fast extensor digitorum longus muscles of wild-type control and dystrophic mdx mice to test the hypothesis that dystrophin deficiency would affect the depolarization-induced contractile response (DICR) and sarcoplasmic reticulum (SR) function. DICR was similar in muscle fibers from mdx and control mice, indicating normal voltage regulation of Ca2+ release. Nevertheless, rundown of DICR (<50% of initial) was reached more rapidly in fibers from mdx than control mice [control: 32 +/- 5 depolarizations (n = 14 fibers) vs. mdx: 18 +/- 1 depolarizations (n = 7) before rundown, P < 0.05]. The repriming rate for DICRs was decreased in fibers from mdx mice, with lower submaximal DICR observed after 5, 10, and 20 s of repriming compared with fibers from control mice (P < 0.05). SR Ca2+ reloading was not different in fibers from control and mdx mice, and no difference was observed in SR Ca2+ leak. Caffeine (2-7 mM)-induced contraction was diminished in fibers from mdx mice compared with control (P < 0.05), indicating depressed SR Ca2+ release channel activity. Our findings indicate that fast fibers from mdx mice exhibit some impairment in the events mediating E-C coupling and SR Ca2+ release channel activity. PMID- 12724138 TI - Role of MDR1 and MRP1 in trophoblast cells, elucidated using retroviral gene transfer. AB - Natural differences in expression and retroviral transduction techniques were used to test the hypothesis that MDR1 P-glycoprotein (P-gp) and MRP1 (multidrug resistance-related protein) contribute to xenobiotic handling by placental trophoblast. RT-PCR and Western blotting in placenta, primary cytotrophoblast cell cultures, and BeWo, JAr, and JEG choriocarcinoma cell lines showed that MRP1 was ubiquitously expressed, whereas MDR1 was absent or minimally expressed in BeWo and JEG cell lines. In syncytiotrophoblast, P-gp was localized predominantly to the microvillous, maternal facing plasma membrane, and MRP1 to the basal, fetal facing plasma membrane. Functional studies showed that cyclosporin A sensitive accumulation of [3H]vinblastine by cells containing both transport proteins was significantly different from those expressing predominantly MRP1. Retroviral gene transfer of MDR1 to BeWo cells confirmed that this difference was due to the relative expression of MDR1. Therefore, both P-gp and MRP1 contribute to xenobiotic handling by the trophoblast. Localization of P-gp to the microvillous membrane suggests an essential role in preventing xenobiotic accumulation by the syncytiotrophoblast and, therefore, in protecting the fetus. PMID- 12724139 TI - Uptake of histamine by mouse peritoneal macrophages and a macrophage cell line, RAW264.7. AB - We have previously demonstrated that dietary histamine is accumulated in the spleens of l-histidine decarboxylase (HDC)-deficient mice, which lack endogenous histamine synthesis. To characterize the clearance system for dietary histamine in mice, we investigated the cell type and mechanism responsible for histamine uptake in the spleens of HDC-deficient mice. Immunohistochemical analyses using an antihistamine antibody indicated that a portion of the CD14+ cells in the spleen is involved in histamine storage. Peritoneal macrophages obtained from Balb/c mice and a mouse macrophage cell line, RAW264.7, had potential for histamine uptake, which was characterized by a low affinity and high capacity for histamine. The histamine uptake by RAW264.7 cells was observed at physiological temperature and was potently inhibited by pyrilamine, chlorpromazine, quinidine, and chloroquine, moderately inhibited by N alpha-methylhistamine, dopamine, and serotonin, and not affected by tetraethylammonium and 1-methyl-4 phenylpyridinium. Intracellular histamine was not metabolized in RAW264.7 cells and was released at physiological temperature in the absence of extracellular histamine. These results suggest that histamine uptake by macrophages may be involved in the clearance of histamine in the local histamine-enriched environment. PMID- 12724140 TI - Glutathione reverses early effects of glycation on myosin function. AB - Nonenzymatic glycosylation (glycation) has been recognized as an important posttranslational modification underlying alterations of structure and function of extracellular proteins during aging and diabetes. Intracellular proteins may also be affected by this modification, and glycation has been suggested to contribute to aging-related impairment in skeletal muscle function. Glycation is the chemical reaction of reducing sugars with primary amino groups resulting in the formation of irreversible advanced glycation end products. Glutathione is an abundant tripeptide in skeletal muscle. To understand the effect of glutathione on glycated myosin function, we used a single-fiber in vitro motility assay in which myosin is extracted from a single muscle fiber segment to propel fluorescent-labeled actin filaments. Myosin function responded to glucose exposure in a dose-dependent manner, i.e., motility speeds were reduced by 10, 34, and 90% of preincubation values after 30-min exposure to 1, 3, and 6 mM glucose, respectively. The 30-min 6 mM glucose incubation was followed by a 20 min 10 mM glutathione incubation. Glutathione treatment restored motility (0.98 +/- 0.06 microm/s, n = 3; P < 0.001) after glucose exposure (0.10 +/- 0.07 microm/s, n = 3), close to preincubation levels (1.12 +/- 0.06 microm/s, n = 3). It is concluded that glucose modifies myosin function in a dose-dependent manner and that glutathione reverses the effect of glucose on myosin function. PMID- 12724141 TI - Control of ascorbic acid efflux in rat luteal cells: role of intracellular calcium and oxygen radicals. AB - In luteal cells, prostaglandin (PG)F2a mobilizes intracellular calcium concentration ([Ca]i), generates reactive oxygen species (ROS), depletes ascorbic acid (AA) levels, inhibits steroidogenesis, and ultimately induces cell death. We investigated the hypothesis that [Ca]i mobilization stimulates ROS, which results in depletion of cellular AA in rat luteal cells. We used a self-referencing AA selective electrode that noninvasively measures AA flux at the extended boundary layer of single cells and fluorescence microscopy with fura 2 and dichlorofluorescein diacetate (DCF-DA) to measure [Ca]i and ROS, respectively. Menadione, a generator of intracellular superoxide radical (O2-), PGF2a, and calcium ionophore were shown to increase [Ca]i and stimulate intracellular ROS. With calcium ionophore and PGF2a, but not menadione, the generation of ROS was dependent on extracellular calcium influx. In unstimulated cells there was a net efflux of AA of 121.5 +/- 20.3 fmol x cm-1 x s-1 (mean +/- SE, n = 8), but in the absence of extracellular calcium the efflux was significantly reduced (10.3 +/- 4.9 fmol x cm-1 x s-1; n = 5, P < 0.05). PGF2a and menadione stimulated AA efflux, but calcium ionophore had no significant effect. These data suggest two AA regulatory mechanisms: Under basal conditions, AA efflux is calcium dependent and may represent recycling and maintenance of an antioxidant AA gradient at the plasma membrane. Under luteolytic hormone and/or oxidative stress, AA efflux is stimulated that is independent of extracellular calcium influx or generation of ROS. Although site-specific mobilization of calcium pools and ROS cannot be ruled out, the release of AA by PGF2a-stimulated luteal cells may occur through other signaling pathways. PMID- 12724142 TI - Functional properties of four splice variants of a human pancreatic tandem-pore K+ channel, TALK-1. AB - TALK-1a, originally isolated from human pancreas, is a member of the tandem-pore K+ channel family. We identified and characterized three novel splice variants of TALK-1 from human pancreas. The cDNAs of TALK-1b, TALK-1c, and TALK-1d encode putative proteins of 294, 322, and 262 amino acids, respectively. TALK-1a and TALK-1b possessed all four transmembrane segments, whereas TALK-1c and TALK-1d lacked the fourth transmembrane domain because of deletion of exon 5. Northern blot analysis showed that among the 15 tissues examined, TALK-1 was expressed mainly in the pancreas. TALK-1a and TALK-1b, but not TALK-1c and TALK-1d, could be functionally expressed in COS-7 cells. Like TALK-1a, TALK-1b was a K+ selective channel that was active at rest. Single-channel openings of TALK-1a and TALK-1b were extremely brief such that the mean open time was <0.2 ms. In symmetrical 150 mM KCl, the apparent single-channel conductances of TALK-1a and TALK-1b were 23 +/- 3 and 21 +/- 2 pS at -60 mV and 11 +/- 2 and 10 +/- 2 pS at +60 mV, respectively. TALK-1b whole cell current was inhibited 31% by 1 mM Ba2+ and 71% by 1 mM quinidine but was not affected by 1 mM tetraethylammonium, 1 mM Cs+, and 100 microM 4-aminopyridine. Similar to TALK-1a, TALK-1b was sensitive to changes in external pH. Acid conditions inhibited and alkaline conditions activated TALK-1a and TALK-1b, with a K1/2 at pH 7.16 and 7.21, respectively. These results indicate that at least two functional TALK-1 variants are present and may serve as background K+ currents in certain cells of the human pancreas. PMID- 12724144 TI - Is there an amygdala and how far does it extend? An anatomical perspective. AB - Burdach is credited with first using the term "amygdala" to describe an almond shaped mass of gray matter in the anterior portion of the human temporal lobe. With the subsequent development of histological techniques in the latter part of the nineteenth century it became readily apparent that there were areas dorsal and medial to Burdach's amygdala that seemed to be anatomically related to it. These regions include the nuclei that are now called the central, medial, and cortical amygdalar nuclei. Johnston considered the amygdala to have two major portions: (1) the centromedial nuclei, which are closely associated with the striatum, and (2) the cortical and basolateral nuclei, which are closely associated with the cerebral cortex. Johnston also importantly observed that the centromedial nuclei appear to have extensions that project rostralward to become continuous with the bed nucleus of the stria terminalis. Johnston's concept of a multicompartmental amygdala, with striatal-like and cortical-like components, dominated thought about the structure and function of the amygdala for most of the twentieth century. Recently, it was suggested that the amygdala is not a single neurobiological entity, but is better conceived as a region where several distinct structures, each with affiliations with different systems, are in close proximity to each other. These current concepts and issues concerning the configuration of the amygdala are discussed in light of recent cytological, immunohistochemical, and connectional studies. PMID- 12724145 TI - Ontogeny of the human amygdala. AB - Data on the fetal development of the human amygdala is reviewed with special reference to major ontogenetic events. In the fifth gestational month, the inferior portion of the amygdala reveals cell-dense columns merging with the ganglionic eminence (proliferative zone) in Nissl-stained sections. These columns contain vimentin-positive fibers and can therefore be regarded as migrational routes. In the sixth and seventh months, distinct reorganization of the cytoarchitectonics takes place. The sequential occurrence of afferens can be visualized using anti-GAP-43; moreover, outgrowing axons appear to reach the periphery of the ganglionic eminence. The latter may thus represent an intermediate target for growing axons using anti-calbindin and anti-calretinin. Migrating and immature amygdaloid neurons can be shown in the fifth month. From the eighth month onwards, various nonpyramidal neurons and pyramidal neurons are immunolabeled. Transient expression of calretinin in pyramidal neurons is observed. When punctate calbindin and calretinin immunostaining in the fifth and eighth months is compared, distinct redistribution is observed. On the whole, it is apparent that the amygdala has reached a high degree of maturity in the eighth month. At this developmental stage, AKAP79, being enriched in postsynaptic densities, shows a characteristic nuclear-specific distribution pattern. The latter largely corresponds to the expression pattern of NMDAR1. Thus, AKAP79 may have a preference for anchoring enzymes to glutamate receptors. The aforementioned results provide a basis for investigations on subtle changes in pathologically altered material, such as hemorrhage, in the ganglionic eminence of preterm infants. PMID- 12724146 TI - Intrinsic synaptic circuitry of the amygdala. AB - The present study is part of an ongoing project aimed at understanding the electrophysiologic properties of single amygdaloid neurons and their correlations with the morphology of the somata as well as axonal and dendritic trees. The axonal morphology of 14 three-dimensional, reconstructed spiny neurons (4 in the lateral and 10 in the basal nucleus) that were filled in vivo with intracellular injections of biocytin is described. Three-dimensional reconstruction was performed using Neurolucida software (MicroBrightField). Sholl analysis was used to assess the axonal length as well as the number of axonal varicosities and endings within concentric spherical shells placed at 50- micro m intervals from the soma. These data indicate that the same neuron can innervate several amygdaloid nuclear divisions or nuclei and extra-amygdaloid regions. This finding suggests that the same neuron can modulate various brain areas in parallel. Both the presumed intra-amygdaloid (all axonal branches within the amygdala) and extra amygdaloid (axons also outside the amygdala) projection neurons have dense perisomatic axonal arborizations, and consequently, the intra-amygdaloid and extra-amygdaloid projection neurons are difficult to differentiate based on the analysis of perisomatic axonal morphology. Furthermore, the same extra-amygdaloid neuron can drive many neurons both locally as well as at extra-amygdaloid projection areas within a relatively short time. Finally, the axonal morphology of spiny neurons located in the lateral or basal nuclei was similar. These data provide baseline quantitative information about the axonal dimensions of amygdaloid neurons and can form the anatomic basis for modeling amygdaloid neuronal circuits when more quantitative data regarding neuronal numbers, size, and dendritic morphology become available. PMID- 12724147 TI - Comparative aspects of amygdala connectivity. AB - The connections of the amygdala in rats, cats, and monkeys can be divided into three systems: (1) A largely forebrain system provides sensory information to the amygdala from the olfactory cortex, ascending taste/visceral pathways, posterior thalamus and sensory association cortex. Most of these connections are reciprocal and also support amygdaloid modulation of sensory processing. (2) A set of projections extends from the amygdala to the hypothalamus and brain stem, which appears to modulate visceral function in relation to emotional stimuli. (3) A forebrain circuit involves the amygdala with the ventromedial frontal, rostral insular, and rostral temporal cortical areas, the medial thalamus and ventromedial basal ganglia, and seems to be directly involved in emotional behavior and mood. All of these systems are present in all mammals studied; the major differences lie in the greater elaboration of the cerebral cortex in primates. PMID- 12724148 TI - Inhibitory and excitatory circuitries in amygdala nuclei: a synopsis of session II. PMID- 12724149 TI - Excitatory synaptic transmission in the lateral and central amygdala. AB - The amygdala plays a major role in the acquisition and expression of fear conditioning. NMDA receptor-dependent synaptic plasticity within the basolateral amygdala has been proposed to underlie the acquisition and possible storage of fear memories. Here the properties of fast glutamatergic transmission in the lateral and central nuclei of the amygdala are presented. In the lateral amygdala, two types of neurons, interneurons and projection neurons, could be distinguished by their different firing properties. Glutamatergic inputs to interneurons activated AMPA receptors with inwardly rectifying current-voltage relations (I-Vs), whereas inputs to projection neurons activated receptors that had linear I-Vs, indicating that receptors on interneurons lack GluR2 subunits. Inputs to projection neurons formed dual component synapses with both AMPA and NMDA components, whereas at inputs to interneurons, the contribution of NMDA receptors was very small. Neurons in the central amygdala received dual component glutamatergic inputs that activated AMPA receptors with linear I-Vs. NMDA receptor-mediated EPSCs had slow decay time constants in the central nucleus. Application of NR2B selective blockers ifenprodil or CP-101,606 blocked NMDA EPSCs by 70% in the central nucleus, but only by 30% in the lateral nucleus. These data show that the distribution of glutamatergic receptors on amygdalar neurons is not uniform. In the lateral amygdala, interneurons and pyramidal neurons express AMPA receptors with different subunit compositions. Synapses in the central nucleus activate NMDA receptors that contain NR1 and NR2B subunits, whereas synapses in the lateral nucleus contain receptors with both NR2A and NR2B subunits. PMID- 12724150 TI - Contextual inhibitory gating of impulse traffic in the intra-amygdaloid network. AB - New data on the organization of the intra-amygdaloid circuit is reviewed, beginning with the basolateral (BL) complex, the main input station of the amygdala for sensory afferents, and concluding with the central (CE) nucleus, an important source of projections to brain-stem structures mediating fear responses. The BL complex is endowed with a highly divergent system of intrinsic glutamatergic connections. Yet, BL projection cells have unusually low firing rates. This apparent contradiction is explained by the presence of powerful inhibitory pressures in the BL amygdala: (1) interneurons that generate large amplitude inhibitory synaptic potentials and (2) projection cells that express a Ca(2+)-dependent K(+) current that can be activated by subthreshold synaptic inputs. Likewise, excitatory projections from the BL amygdala to the CE nucleus are controlled by clusters of GABAergic neurons, termed the intercalated (ITC) cell masses. In response to BL inputs, ITC cells generate feedforward inhibition in CE neurons. However, ITC neurons exhibit properties that allow them to modify the amount of inhibition they generate depending on the distribution of BL activity in space and time. Indeed, ITC cell masses can inhibit each other via lateromedial connections. Moreover, they express an unusual K(+) conductance that modifies their response to BL inputs depending on their recent firing history. Thus, inhibitory mechanisms of the amygdala allow for flexible, context-dependent gating of BL impulses to the CE nucleus. PMID- 12724151 TI - Genes and mechanisms in the amygdala involved in the formation of fear memory. AB - With a combined molecular, electrophysiological, and behavioral approach we have sought to correlate conditioned fear behavior with electrophysiological activities in the lateral amygdala and hippocampal formation in mice and rats and to determine the potential contribution of effector genes that are expressed in the basolateral amygdaloid complex during the late phase of pavlovian fear conditioning. Our data indicate that resonant/oscillatory electrical activity in projection neurons of the lateral amygdala provide an important cellular element of coherent theta activity in amygdalohippocampal pathways, which may represent a nondiscriminating neural correlate of conditioned fear. Correlated activity seems to contribute to the formation of synaptic plasticity in these networks, such as input-specific long-term depression of thalamoamygdaloid signals and consolidation of long-term potentiation in the dentate gyrus. Moreover, associative fear conditioning results in selective gene expression in the basolateral amygdaloid complex, involving molecular factors of structural reorganization and signal transduction, particularly GABA function, supporting the view that the amygdala is a site of neural plasticity and information storage during formation of fear memory. PMID- 12724152 TI - The amygdala, synaptic plasticity, and fear memory. AB - The nature and mechanisms of synaptic plasticity in the amygdala and the relation of amygdaloid plasticity to behavior are exciting new areas of study in neuroscience. These issues were at the heart of presentations by Paul Chapman, Michael Fanselow, Patricia Shinnick-Gallagher, and Michael Rogawski in a session entitled "Long-Term Plasticity in Amygdala Synaptic Transmission" that was held at the conference featured in this volume. In this chapter, I briefly summarize these talks and give my perspective on the presentations as the session chair. I argue that we must first understand the role of the amygdala in learning and memory in order to understand the contribution of amygdaloid synaptic plasticity to behavior. Although it is generally agreed that the amygdala is involved in several forms of emotional learning and memory such as pavlovian fear conditioning, a recent debate has emerged concerning the precise role of the amygdala in learning versus performing fear responses. I discuss data from my laboratory that unravel this issue. I argue that the basolateral complex of the amygdala (BLA) normally plays an essential role in associative processes in fear conditioning. Nonetheless, rats with BLA lesions acquire and express conditional fear under some conditions. A neuroanatomical model that accounts for these data is presented. PMID- 12724153 TI - Synaptic plasticity in the amygdala: comparisons with hippocampus. AB - Long-term potentiation (LTP) is a widely studied form of synaptic plasticity, and a considerable amount of evidence indicates that it could be involved in learning and memory. Intensive investigation of this phenomenon in the hippocampus has yielded tremendous insight into the workings of synapses in the mammalian central nervous system, but important questions remain to be answered. The most important of these are: (1) whether LTP is the basis of learning and memory, and (2) how similar are the induction, maintenance, and expression mechanisms in the rest of the brain to those in the hippocampus. Because the most important strategy for linking LTP to learning involves disrupting the mechanisms of LTP and examining the consequences on behavior, it is likely that the first question cannot be answered until the second has been addressed. Recent evidence indicates that although the general processes have much in common, significant differences exist among forebrain structures, including the hippocampus, basolateral amygdala, and ventral striatum. It is clear that the roles of receptors and calcium channels, kinases, and transcription factors vary within these structures, reflecting the different functions of these brain regions. PMID- 12724154 TI - The amygdala, fear, and memory. AB - Lesions of the frontotemporal region of the amygdala, which includes lateral and basal nuclei, cause a loss of conditional fear responses, such as freezing, even when the lesions are made over a year and a half from the original training. These amygdala-damaged animals are not hyperactive and show normal reactivity to strong stimuli such as bright lights. After receiving tone-mild shock pairings rats normally display an appropriately weak response when exposed to the tone. Rats' fear of the tone can be inflated by giving them exposure to strong shocks in the absence of the tone between training and testing. This inflation of fear memory is abolished if the frontotemporal amygdala is inactivated by muscimol only during the inflation treatment with strong shocks. Based on such findings we suggest that the frontotemporal amygdala permanently encodes a memory for the hedonic value of the aversive stimulus used to condition fear. PMID- 12724155 TI - L-type voltage-gated calcium channels are involved in the in vivo and in vitro expression of fear conditioning. AB - Fear conditioning, a behavioral model of fear learning and cue-related anxiety, causes enhanced neuronal transmission in the thalamic to lateral amygdala pathway.(1,2) In the expression phase of learned fear, this increased transmission recorded in vitro is revealed in increased amplitudes of excitatory postsynaptic currents (EPSCs) and occlusion of paired-pulse facilitation (PPF) implicating a presynaptic increase in transmitter release. Here we examined the contribution of L-type calcium channels in fear conditioning. We measured the effect of nimodipine (Nim, 1.5-20 mg/kg), an L-type calcium channel antagonist, on fear-potentiated startle in which startle was assessed in animals receiving paired or unpaired tone and foot shock. Nim administered intraperitoneally blocked fear-potentiated startle but not baseline startle in a dose-dependent manner. We also analyzed the effect of Nim (10 micro M) in vitro on synaptic facilitation of EPSCs and PPF in slices from naive control, unpaired control, and fear-conditioned animals. In neurons from naive control animals, Nim had no effect on EPSC amplitude or PPF, but in slices from fear-conditioned rats, Nim reduced EPSC amplitude, suggesting the recruitment of L-type calcium channels within the fear-conditioning pathway. Nim increased PPF in slices from fear conditioned animals, suggesting that L-type calcium channels may contribute to increased probability of release in fear conditioning. In slices from unpaired animals, Nim decreased synaptic transmission but had little effect on PPF, suggesting that stress or contextual fear learning may induce L-type channel activity in fear-conditioned and unpaired control animal groups. We also analyzed protein expression of the alpha(1C) and alpha(1D) L-type calcium channel subunits isolated from the amygdala and found that alpha(1C) protein was significantly increased in fear-conditioned animals. These findings suggest that L-type calcium channels play a role in the amygdala in cued fear conditioning and have important implications in the treatment of anxiety and in emotional learning and plasticity. PMID- 12724156 TI - GluR5 kainate receptors, seizures, and the amygdala. AB - The amygdala is a critical brain region for limbic seizure activity, but the mechanisms underlying its epileptic susceptibility are obscure. Several lines of evidence implicate GluR5 (GLU(K5)) kainate receptors, a type of ionotropic glutamate receptor, in the amygdala's vulnerability to seizures and epileptogenesis. GluR5 mRNA is abundant in temporal lobe structures including the amygdala. Brain slice recordings indicate that GluR5 kainate receptors mediate a portion of the synaptic excitation of neurons in the rat basolateral amygdala. Whole-cell voltage-clamp studies demonstrate that GluR5 kainate receptor-mediated synaptic currents are inwardly rectifying and are likely to be calcium permeable. Prolonged activation of basolateral amygdala GluR5 kainate receptors results in enduring synaptic facilitation through a calcium-dependent process. The selective GluR5 kainate receptor agonist ATPA induces spontaneous epileptiform bursting that is sensitive to the GluR5 kainate receptor antagonist LY293558. Intra amygdala infusion of ATPA in the rat induces limbic status epilepticus; in some animals, recurrent spontaneous seizures occur for months after the ATPA treatment. Together, these observations indicate that GluR5 kainate receptors have a unique role in triggering epileptiform activity in the amygdala and could participate in long-term plasticity mechanisms that underlie some forms of epileptogenesis. Accordingly, GluR5 kainate receptors represent a potential target for antiepileptic and antiepileptogenic drug treatments. Most antiepileptic drugs do not act through effects on glutamate receptors. However, topiramate at low concentrations causes slow inhibition of GluR5 kainate receptor mediated synaptic currents in the basolateral amygdala, indicating that it may protect against seizures, at least in part, through suppression of GluR5 kainate receptor responses. PMID- 12724157 TI - Sex-related influences on the neurobiology of emotionally influenced memory. AB - In a recent report, the National Academy of Sciences concluded that "Sex matters. Sex, that is, being male or female, is an important basic human variable that should be considered when designing and analyzing studies in all areas and at all levels of biomedical and health-related research."(1) Recent findings from my laboratory concerning neural mechanisms of emotionally influenced memory further support this conclusion. This article first provides a brief, general overview of sex-related influences on brain and cognition. Upon this background, recent findings from my laboratory and others are described, demonstrating sex-related influences on neural mechanisms underlying emotionally influenced, explicit recall of emotionally arousing events. Both the hemispheric involvement of the human amygdala in memory for emotionally arousing events and the impairing effect of beta-adrenergic blockade on memory for emotional events exhibit pronounced sex related differences. We interpret both of these effects in the context of evidence indicating hemispheric specialization in the processing of global/wholistic versus local/fine detail aspects of a situation. The more general conclusion that we draw from these investigations is that theories of the neurobiology of emotion and memory must begin to account for the seemingly substantial influences of sex. PMID- 12724158 TI - The amygdala and its place in the cerebral hemisphere. AB - Developmental, gene expression, connectional, and neuron morphology evidence converges to suggest that cell groups of the amygdalar region participate in at least four distinct, though interrelated, functional systems associated with differentiated corticostriatopallidal projections. The amygdala is neither a structural nor a functional unit; instead, it is a collection of adjacent cell groups in the medial temporal lobe and adjacent regions of the caudal piriform lobe that was defined on gross anatomic terms. PMID- 12724159 TI - Extended amygdala and basal forebrain. AB - The basal forebrain is a confluence of systems that are crucial to understanding some of the most important functions of the brain, including reward and punishment, learning and cognition, and feeding and reproduction. Basic to understanding this broad spectrum of behavior is untangling the interwoven functional systems in basal forebrain. This has been grounded by the appreciation that the major nearby structures, that is, amygdala and basal ganglia, provide a context for interpreting basal forebrain areas that are best viewed as extensions of either of these larger regions. The components of basal forebrain, the ventral striatopallidal system and the medial and central divisions of extended amygdala, are subcortical relays for information garnered from brain stem, thalamus, and cortical areas. With respect to the classically defined amygdala of the temporal lobe, the lateral-basolateral complex, and the superficial amygdaloid nuclei may tentatively be viewed as specialized cortical regions. Their output targets both the striatopallidal complex and the extended amygdala, with some of the most massive basal forebrain efferents originating in the basolateral amygdaloid complex. The subcortical projections of the basolateral nucleus, at least in the rat, appear to be dichotomous, with anterior (or magnocellular) portions of the nucleus preferentially targeting striatum and ventral striatum (including the core of the nucleus accumbens), while the posterior (small-celled) portions of the basolateral nucleus target the extended amygdala as well as the shell of the nucleus accumbens. This divergence represents a particular opportunity for behavioral neuroscientists analyzing basal forebrain functions. Studies exploiting the dual subcortical projection of basolateral amygdala indicate distinct functional roles for striatum versus extended amygdala. These reinforce the identification of extended amygdala as a functional-anatomical entity distinct from the striatopallidal system. PMID- 12724160 TI - Consideration of a unified model of amygdalar associative functions. AB - This paper considers issues arising from presentations by Barry Everitt and Gorica Petrovich concerning amygdalar involvement in appetitive conditioning. Studies reported by these researchers are beginning to unravel the distribution of associative functions among amygdalar nuclei and their efferent targets. The main issue addressed here concerns how to integrate the information from studies of appetitive conditioning with conclusions from studies of other forms of associative learning, including pavlovian fear conditioning and instrumental conditioning. Most students of this issue will agree that no prima facie integration is readily apparent in current scholarship. Although we do not here provide the definitive integration, we do attempt to take some preliminary, broad brush steps towards that goal. PMID- 12724161 TI - Role of the amygdala in fear extinction measured with potentiated startle. AB - Although much is now known about the neural basis of excitatory fear conditioning, much less is known about the neural basis of inhibitory conditioning. One type of inhibitory conditioning is extinction, a process in which stimuli that elicit fear by virtue of previous associations with aversive stimuli such as shock (excitatory fear conditioning) are now presented in the absence of the aversive stimuli (extinction training). As a result, the ability of the conditioned stimulus to elicit fear gradually diminishes. Extinction is different from forgetting and does not reflect an erasure of the original fear memory. Instead, extinction is an active form of inhibitory learning that competes with excitatory fear conditioning. Infusions into the amygdala (a brain area essential for excitatory fear conditioning) of either NMDA receptor antagonists or inhibitors of the NMDA-receptor-linked mitogen-activated protein kinase cascade block extinction learning. Conversely, the NMDA receptor agonist D cycloserine facilitates extinction after either systemic administration or intra amygdala infusion. Because therapeutic interventions based on extinction procedures are commonly used to treat fear disorders, and because D-cycloserine is a widely available and safe compound, D-cycloserine or similar agents might be usefully combined with traditional extinction-based approaches in the treatment of clinical fear. PMID- 12724162 TI - Appetitive behavior: impact of amygdala-dependent mechanisms of emotional learning. AB - In this chapter, we review data from studies involving appetitive conditioning using measures of pavlovian approach behavior and the effects of pavlovian conditioned stimuli on instrumental behavior, including the pavlovian-to instrumental transfer effect and conditioned reinforcement. These studies consistently demonstrate double dissociations of function between the basolateral area and the central nucleus of the amygdala. Moreover, the data show marked parallels with data derived from aversive (fear) conditioning studies and are consistent with the idea that these subsystems of the amygdala mediate different kinds of associative representation formed during pavlovian conditioning. We hypothesize that the basolateral amygdala is required for a conditioned stimulus to gain access to the current affective value of its specific unconditioned stimulus, whereas the central nucleus mediates stimulus-response representations and conditioned motivational influences on behavior. Although these systems normally operate together, they can also modulate behavior in distinct ways. In many circumstances, then, emotional behavior can be seen as a coordinated combination of processing by these amygdaloid sub-nuclei, reflecting the superimposition of a phylogenetically recent basolateral amygdala subsystem that encodes and retrieves the affective value of environmental stimuli and thereby directs complex, adaptive behavioral responses onto a phylogenetically older central amygdala subsystem that enables cortical structures (including the basolateral amygdala) to recruit incentive motivational processes and thereby invigorate emotional responding. PMID- 12724163 TI - Amygdala subsystems and control of feeding behavior by learned cues. AB - A combination of behavioral studies and a neural systems analysis approach has proven fruitful in defining the role of the amygdala complex and associated circuits in fear conditioning. The evidence presented in this chapter suggests that this approach is also informative in the study of other adaptive functions that involve the amygdala. In this chapter we present a novel model to study learning in an appetitive context. Furthermore, we demonstrate that long recognized connections between the amygdala and the hypothalamus play a crucial role in allowing learning to modulate feeding behavior. In the first part we describe a behavioral model for motivational learning. In this model a cue that acquires motivational properties through pairings with food delivery when an animal is hungry can override satiety and promote eating in sated rats. Next, we present evidence that a specific amygdala subsystem (basolateral area) is responsible for allowing such learned cues to control eating (override satiety and promote eating in sated rats). We also show that basolateral amygdala mediates these actions via connectivity with the lateral hypothalamus. Lastly, we present evidence that the amygdalohypothalamic system is specific for the control of eating by learned motivational cues, as it does not mediate another function that depends on intact basolateral amygdala, namely, the ability of a conditioned cue to support new learning based on its acquired value. Knowledge about neural systems through which food-associated cues specifically control feeding behavior provides a defined model for the study of learning. In addition, this model may be informative for understanding mechanisms of maladaptive aspects of learned control of eating that contribute to eating disorders and more moderate forms of overeating. PMID- 12724164 TI - Inhibition of the amygdala: key to pathological states? AB - The amygdala plays an important role in associating sensory stimuli with aversive or appetitive outcomes. Conditioning procedures potentiate inputs to the amygdala, which facilitate emotional responses via subcortical and cortical outputs. Powerful inhibitory circuits exist that control expression of conditioned responses in the amygdala, including inhibition from prefrontal cortex. Deficient inhibitory tone in the amygdala could lead to overexpression of conditioned responses, producing pathological states such as anxiety disorders and drug-seeking behavior. Support for this comes from several lines of animal research: (1) GABA antagonist-induced priming of anxiety states, (2) extinction of conditioned fear, (3) modulation of inhibitory avoidance memory, and (4) cue induced reinstatement of drug seeking. Cue-induced craving in humans is associated with feelings of fear and autonomic arousal, suggesting a link between fear and addiction in the amygdala. Future therapies aimed at increasing inhibitory tone in the amygdala, either locally or via the prefrontal cortex, may prevent anxiety disorders and addiction relapse. Novel neuropeptides, which can either excite or inhibit specific components of anxiety responses, offer promise in this regard. PMID- 12724165 TI - Role of the basolateral amygdala in memory consolidation. AB - Memories of emotionally arousing events tend to be more vivid and to persist longer than do memories of neutral or trivial events. Moreover, memories of emotionally influenced information may endure after a single experience. Recent findings strongly suggest that the influence of emotional arousal on memory consolidation is mediated by the release of adrenal stress hormones (epinephrine and glucocorticoids) and neurotransmitters that converge in modulating the noradrenergic system within the amygdala. Considerable evidence also indicates that amygdala activation influences memory by regulating consolidation in other brain regions. The findings suggest further that this memory-modulatory system may be involved in the formation of traumatic memories and posttraumatic stress disorder in human subjects. PMID- 12724166 TI - Drug addiction, relapse, and the amygdala. AB - Evidence has extensively implicated the amygdala in the associative learning process for appetitive reinforcers. Recent interest has focused on the role of the amygdala in the learned associations that occur during the process of drug addiction and relapse. Using an animal model of relapse after chronic cocaine self-administration, we found that rats reinstate extinguished lever responding for conditioned stimuli (tone + light) previously paired with cocaine or heroin ("conditioned-cued reinstatement"). The basolateral amygdala (BLA) complex plays a critical role in this behavior, because permanent lesions or reversible pharmacologic inactivation of the BLA attenuates conditioned-cued reinstatement without affecting cocaine self-administration or cocaine-primed reinstatement. Conditioned-cued reinstatement appears to be mediated in part by dopamine inputs to the BLA, as intra-BLA infusion of a dopamine D1 receptor antagonist blocks reinstatement, whereas intra-BLA infusion of amphetamine potentiates reinstatement. Furthermore, the BLA is also necessary for acquisition of associative learning with cocaine-paired stimuli. Disruption of neural activity within the BLA by sodium channel blockade or muscarinic receptor blockade just before acquisition of stimulus-cocaine associations blocks the ability of conditioned stimuli to elicit conditioned-cued reinstatement after extinction. Together, these results reveal the importance of the amygdala as part of a corticolimbic circuit mediating both the acquisition and the expression of conditioning that plays a critical role in relapse to drug-seeking behavior. PMID- 12724167 TI - The amygdala, panic disorder, and cardiovascular responses. AB - The amygdala is implicated in a number of emotional responses including conditioned fear and anxiety, and it appears to regulate the behavioral and autonomic responses associated with such emotional responses. The basolateral nucleus of the amygdala (BLA) is under tonic GABAergic inhibition, and acutely blocking this inhibition results in increased anxiety-like behavior, conditioned avoidance, and sympathetically mediated cardiovascular activation. By contrast, activation of the BLA with the stress-related neuropeptide corticotropin releasing factor results in anxiety-like behavior, but not cardiovascular activation. Furthermore, repeated activation of this region with subthreshold GABA blockade or corticotropin-releasing factor-mediated excitation (priming) results in a chronic anxiety-like state, with susceptibility to panic-like arousal following intravenous lactate infusions. The chronic anxiety state appears to result from a loss of basal inhibitory drive in the BLA as a result of NMDA-dependent synaptic plasticity involving cyclic AMP and calcium calmodulin kinase II (CAM KII)-mediated changes. The lactate-induced panic-like response appears to involve angiotensin-II mediated activation of the BLA. These results suggest that the BLA has a significant role in regulating anxiety, autonomic responses, and the development of anxiety disorders. PMID- 12724168 TI - Is the human amygdala specialized for processing social information? AB - A number of studies in humans and other animals has confirmed the amygdala's role in modulating cognition and behavior on the basis of a stimulus' motivational, emotional, and social attributes. This raises the question of how these attributes are related: is social information processing reducible to motivational processing? Some recent data suggest the possibility that the amygdala's primitive function may be motivational processing that is domain general, but that its function in primates, and especially humans, may have evolved to process social information specifically. While the issue is unresolved, future experiments could provide additional support. PMID- 12724169 TI - Amygdaloid regional cerebral blood flow and subjective fear during symptom provocation in anxiety disorders. AB - Whether the amygdala is involved predominantly in emotional perception or in the generation of emotional states has been debated. We reviewed and reanalyzed data from our laboratory, indicating that subjective feelings of fear and distress are correlated with regional cerebral blood flow (rCBF) in the right but not the left amygdala during anxiety provocation in individuals with social anxiety disorder, specific phobias. and posttraumatic stress disorder. Positron emission tomography is a correlative technique, and casual inferences cannot be drawn. However, because studies demonstrate that treatment of social anxiety disorder with cognitive behavior therapy and selective serotonin reuptake inhibitors results in reduced rCBF in the amygdaloid complex and prospective studies reveal that treatment-induced alterations in amygdala rCBF can predict 1 year follow-up status in social anxiety disorder data support the notion that the amygdala, at least in part, seem casually involved in generating the subjective experience of fear. PMID- 12724170 TI - Amygdala automaticity in emotional processing. AB - The question addressed in this paper is whether the human amygdala processes threat-related stimuli independent of selective attention. This is considered from a functional neuroimaging perspective, describing studies in normal volunteers and patients with brain lesions. The conclusion is that the evidence for such automaticity is strong, indicating that important distinctions exist between emotion and other forms of cognition. PMID- 12724171 TI - Role of the amygdala in decision-making. AB - The somatic marker hypothesis proposes that both the amygdala and the orbitofrontal cortex are parts of a neural circuit critical for judgment and decision-making. Although both structures couple exteroceptive sensory information with interoceptive information concerning somatic/emotional states, they do so at different levels, thus making different contributions to the process. We define "primary inducers" as stimuli that unconditionally, or through learning (e.g., conditioning and semantic knowledge), can (perceptually or subliminally) produce states that are pleasurable or aversive. Encountering a fear object (e.g., a snake), a stimulus predictive of a snake, or semantic information such as winning or losing a large sum of money are all examples of primary inducers. "Secondary inducers" are entities generated by the recall of a personal or hypothetical emotional event or perceiving a primary inducer that generates "thoughts" and "memories" about the inducer, all of which, when they are brought to memory, elicit a somatic state. The episodic memory of encountering a snake, losing a large sum of money, imagining the gain of a large sum of money, or hearing or looking at primary inducers that bring to memory "thoughts" pertaining to an emotional event are all examples of secondary inducers. We present evidence in support of the hypothesis that the amygdala is a critical substrate in the neural system necessary for triggering somatic states from primary inducers. The ventromedial cortex is a critical substrate in the neural system necessary for the triggering of somatic states from secondary inducers. The amygdala system is a priori a necessary step for the normal development of the orbitofrontal system for triggering somatic states from secondary inducers. However, once this orbitofrontal system is developed, the induction of somatic states by secondary inducers via the orbitofrontal system is less dependent on the amygdala system. Perhaps the amygdala is equivalent to the hippocampus with regard to emotions, that is, necessary for acquiring new emotional attributes (anterograde emotions), but not for retrieving old emotional attributes (retrograde emotions). Given the numerous lesion and functional neuroimaging studies illustrating the involvement of the amygdala in complex cognitive and behavioral functions, including "social cognition," we suggest that this involvement is a manifestation of a more fundamental function mediated by the amygdala, which is to couple stimuli/entities with their emotional attributes, that is, the processing of somatic states from primary inducers. PMID- 12724172 TI - Brain imaging studies in mood and anxiety disorders: special emphasis on the amygdala. AB - Human studies attempting to elucidate brain functioning in health and disease are crucial for our understanding of neuropsychiatric disorders. In the past, scientists relied heavily on neurological lesion studies to understand the functional roles of brain areas. In the last few decades, brain imaging research has made it possible to investigate the molecular and synaptic neuronal events as well as the functioning of neuronal networks in vivo, in patients with neuropsychiatric illnesses. In this context, the functional role of the amygdala has been a focus of neuroimaging studies by leading researchers. Several of these researchers presented papers at a conference, entitled The Amygdala in Brain Function: Basic and Clinical Approaches, that provided the basis for this volume. These papers follow this review in the current volume. The present paper briefly summarizes the highlights of the different presentations, focusing on the functional diversity of the amygdala and its role in different neuropsychiatric disorders; reviews the various brain imaging technologies currently available; and discusses the major findings on the pathophysiology and treatment of depression, bipolar disorder, and anxiety disorders. PMID- 12724173 TI - Neuroimaging studies of amygdala function in anxiety disorders. AB - Neuroimaging research has helped to advance neurobiological models of anxiety disorders. The amygdala is known to play an important role in normal fear conditioning and is implicated in the pathophysiology of anxiety disorders. The amygdala may also be a target for the beneficial effects of cognitive-behavioral and medication treatments for anxiety disorders. In the current paper, we review neuroimaging research pertaining to the role of the amygdala in anxiety disorders and their treatment. Moreover, we discuss the development of new neuroimaging paradigms for measuring aspects of amygdala function, as well as the function of related brain regions. We conclude that such tools hold great promise for facilitating progress in relevant basic neuroscience as well as clinical research domains. PMID- 12724174 TI - The amygdala and Alzheimer's disease: positron emission tomographic study of the cholinergic system. AB - The primary transmitter deficit is cholinergic in Alzheimer's disease (AD), and the amygdala receives a major cholinergic projection from the nucleus basalis of Meynert (Ch4), which may play an important role in the retention of affective conditioning and/or memory consolidation. We measured brain acetylcholinesterase (AChE) activity in 54 patients with AD and in 22 normal controls by positron emission tomography and N-[(11)C]methylpiperidin-4-yl acetate to characterize the cholinergic pathology in AD. The k(3) values were calculated as an index of AChE activity in a three-compartment model analysis using the metabolite-corrected arterial input function. The k(3) values were highly significantly reduced by 20% in the cerebral neocortex (P <0.0001 in the two-tailed t test), 14% in the hippocampus (P <0.001), and 33% in the amygdala (P <0.0001) in AD patients compared with normal controls. The k(3) values were significantly correlated with the Mini-Mental State Examination scores in both the cerebral cortex (P <0.001) and the amygdala (P <0.05) in AD patients, supporting the cholinergic hypothesis of cognitive dysfuncion in AD. Further studies are required, however, to elucidate the specific role of the cholinergic deficit in the amygdala in the emotional and behavioral symptoms in AD. PMID- 12724175 TI - Neuroimaging abnormalities in the amygdala in mood disorders. AB - Neuroimaging technology has been applied to investigate the pathophysiology of mood disorders in studies aimed at characterizing the anatomical correlates of depressive symptoms, the neurophysiological effects of antidepressant treatments, and the trait-like abnormalities that persist despite symptom remission. These studies have identified cerebral blood flow and metabolic differences between depressives and controls in the amygdala and anatomically related areas of the prefrontal cortex, striatum, and thalamus. Taken together with converging evidence from neuroendocrine, lesion analysis, and postmortem studies of clinically depressed subjects, these data suggest that emotional/stress-response systems that include the amygdala are pathologically activated in major depression and that this activity is associated with dysfunction of the prefrontal cortex and monoamine neurotransmitter systems that normally modulate such responses. PMID- 12724176 TI - Structural and functional abnormalities of the amygdala in schizophrenia. AB - Schizophrenia is characterized by delusions and hallucinations, which tend to respond to treatment with dopamine receptor blockers, and a loss of motivation and affect, which do not. Structural magnetic resonance imaging (sMRI) has convincingly demonstrated reduced volumes of the amygdala-hippocampal complex (AHC) and other limbic and paralimbic structures, on both manual tracing and automated analyses. The Edinburgh High-Risk Study (EHRS) of initially healthy adolescents with at least two affected relatives has found that AHC volumes are reduced pre-morbidly but not to schizophrenic levels, suggesting that further volume reductions may be associated with the onset of schizophrenia. AHC volumes appear to be genetically mediated in families with a dominant pattern of transmission, whereas prefrontal lobe and basal ganglia volumes are related to genetic liability to schizophrenia in the generality of high-risk subjects. Temporal lobe volumes may fall as psychotic symptoms develop, in the context of drug abuse and stress. Neuropsychological testing has also demonstrated pre morbid impairments and symptom-related deterioration. More detailed analyses of the temporal lobe changes on sMRI and fronto-temporal dysconnectivity on fMRI are in progress. These findings are discussed with reference to other indications of pre-morbid developmental disturbance in our high-risk subjects, animal models of schizophrenia, and reliable findings from neuropathological, neuropsychological, and functional imaging studies of patients with schizophrenia. PMID- 12724177 TI - The amygdala and psychopathology studies in epilepsy. AB - The results of volumetric studies of the amygdala and hippocampus in patients with epilepsy who also have psychopathology are described. The main data relate to the increased size of the amygdala in patients with epilepsy and psychosis. The associations between the amygdala and affective disorders, notably dysthymia and euphoria, are also discussed. PMID- 12724178 TI - The amygdala in brain function: the Datablitz sessions. PMID- 12724179 TI - Effect of amygdalar opioids on the anxiolytic properties of ethanol. PMID- 12724180 TI - Amygdalar recruitment during anticipation of monetary rewards: an event-related fMRI study. PMID- 12724181 TI - Effects of chronic alcohol ingestion on rat lateral/basolateral amygdala ligand gated chloride channels. PMID- 12724182 TI - Relationships between amygdala volume and activity during emotional information processing tasks in depressed and never-depressed individuals: an fMRI investigation. PMID- 12724183 TI - Associative plasticity in neurons of the lateral amygdala during auditory fear conditioning. PMID- 12724207 TI - Epigenetics in cancer prevention: early detection and risk assessment: introduction. PMID- 12724184 TI - Affective conditioning in the basolateral amygdala of anesthetized rats is modulated by dopamine and prefrontal cortical inputs. PMID- 12724208 TI - Chromatin as a tool for the study of genome function in cancer. AB - The transition to malignancy requires an extensive reconfiguration of the genome's expression program that does not result entirely from actual changes in primary DNA sequence-i.e., mutation. Epigenetic-meta-DNA-gene expression states result from an assembly over a given locus of a poorly understood nucleoprotein entity that includes histones and other architectural components of chromatin, nonhistone DNA-bound regulators, and additional chromatin-bound polypeptides. This structure is rapidly reestablished in the wake of the DNA replication fork, thus ensuring its persistence in rapidly proliferating cells and thereby yielding an exceptionally stable mode of gene expression. Chromatin is the perfect vehicle for enabling such genome control. During S phase both covalently modified histones and histone-associated regulatory proteins distribute to the newly synthesized daughter chromatids in a form of "molecular dowry" inherited from the G(1) state of the genome, and impose a specific mode of function on the underlying DNA. An extensively studied example of chromatin-based epigenetic inheritance connects DNA methylation to the targeting of chromatin remodeling and modification. In a broad sense, however, genome reprogramming in cancer is associated with the remodeling of a multitude of regulatory DNA stretches-e.g., promoters, enhancers, locus control regions (LCRs), insulators, etc.-into a specific chromatin architecture. This architectural entity provides a general molecular signature of the cancer epigenome that complements and significantly expands its DNA methylation-based component. PMID- 12724209 TI - Unanswered questions about the role of promoter methylation in carcinogenesis. AB - It has been proposed that tumor suppressor genes can be silenced by ectopic de novo methylation during tumor progression and that this epigenetic silencing is an alternative to mutation in tumor suppressor inactivation during oncogenic transformation. However, methylation may follow inactivation and may not directly participate in tumor progression. There are no genetic data that implicate ectopic de novo methylation in cancer, and no DNA methyltransferase gene has been shown to be mutated in any cancer. Promoter methylation at tumor suppressor loci may be a consequence of transcriptional inactivity imposed by mutations in upstream components of the transcriptional machinery or signal transduction pathways. Current estimates of the importance of epigenetic changes in the etiology of cancer may be inflated, and consequences may have been mistaken for causes in some cases. PMID- 12724210 TI - Hypomethylation: one side of a larger picture. AB - Hypomethylation signifies one end of a spectrum of DNA methylation states. In most cases hypomethylation refers to a relative state that represents a change from the "normal" methylation level. Hypomethylation, when approached from a topographical perspective, has been used to describe either overall decreases in the methylation status of the entire genome (global hypomethylation) or more localized relative demethylation of specific subsets of the genome, such as the promoter regions of protooncogenes or normally highly methylated repetitive sequences. Global hypomethylation accompanied by gene-specific hypermethylation is observed in at least two important settings: cancer and aging. Global hypomethylation is generally reflective of decreased methylation in CpGs dispersed throughout repetitive sequences as well as the bodies of genes. Hypomethylation of repetitive and parasitic DNA sequences correlates with a number of adverse outcomes. For example, decreased methylation of repetitive sequences in the satellite DNA of the pericentric region of chromosomes is associated with increased chromosomal rearrangements, a hallmark of cancer. Decreased methylation of proviral sequences can lead to reactivation and increased infectivity. However, hypomethylation in cancer can also affect the CpGs in the promoters of specific genes-namely, protooncogenes-leading to their overexpression and resulting in the functional outcome of increased cell proliferation. Thus, hypomethylation, in a variety of settings in which it represents a deviation from "normal," appears to correlate with progression to cancer and offers potential mechanisms to explain the carcinogenic process. PMID- 12724211 TI - Genetic and epigenetic mosaicism in cancer precursor tissues. AB - The concept of field effects in cancer is old, but recent molecular data have substantiated it. Clones of cells that carry well-defined genetic or epigenetic aberrations, but which have not yet acquired the morphological hallmarks of neoplasia, have been documented in the precursor tissues of some of the most common pediatric and adult malignancies. Here I review this evidence, focusing on loss of heterozygosity (LOH) and gain of DNA methylation. PMID- 12724213 TI - Epigenetics: the role of methylation in the mechanism of action of tumor suppressor genes. AB - Epigenetics is the study of mitotically heritable changes in gene expression without any changes in the primary DNA sequence. The major step in epigenetic gene regulation is gene inactivation by hypermethylation of CpG islands located in the promoter region. Specific enzymes and methylated DNA binding proteins play a major role in causing reduced expression of tumor suppressor genes, resulting in tumor formation and its progression. Prevention approaches are needed to avoid tumor formation. One approach to inhibiting inactivation of tumor suppressor genes is to use chemical agents such as 5-azacytidine to prevent hypermethylation of DNA. Increased understanding of the mechanism of epigenetic silencing and the identification of additional molecular mechanisms (e.g., histone methylases) that may be targeted by pharmaceutical interventions may lead to more preventive strategies. The current status of the epigenetic regulation of tumor suppressor genes is discussed in this review article. PMID- 12724212 TI - Epigenetic control during lymphoid development and immune responses: aberrant regulation, viruses, and cancer. AB - Methylation of cytosines controls a number of biologic processes such as imprinting and X chromosomal inactivation. DNA hypermethylation is closely associated with transcriptional silencing, while DNA hypomethylation is associated with transcriptional activation. Hypoacetylation of histones leads to compact chromatin with reduced accessibility to the transcriptional machinery. Methyl-CpG binding proteins can recruit corepressors and histone deacetylases; thus, the interplay between these epigenetic mechanisms regulates gene activation. Methylation has been implicated as an important mechanism during immune development, controlling VDJ recombination, lineage-specific expression of cell surface antigens, and transcriptional regulation of cytokine genes during immune responses. Aberrations in epigenetic machinery, either by genetic mutations or by somatic changes such as viral infections, are associated with early alterations in chronic diseases such as immunodeficiency and cancer. PMID- 12724214 TI - Histone deacetylases: unique players in shaping the epigenetic histone code. AB - The epigenome is defined by DNA methylation patterns and the associated posttranslational modifications of histones. This histone code determines the expression status of individual genes dependent upon their localization on the chromatin. The silencing of gene expression is associated with deacetylated histones, which are often found to be associated with regions of DNA methylation as well as methylation at the lysine 4 residue of histone 3. In contrast, the activation of gene expression is associated with acetylated histones and methylation at the lysine 9 residue of histone 3. The histone deactylases play a major role in keeping the balance between the acetylated and deacetylated states of chromatin. Histone deacetylases (HDACs) are divided into three classes: class I HDACs (HDACs 1, 2, 3, and 8) are similar to the yeast RPD3 protein and localize to the nucleus; class II HDACs (HDACs 4, 5, 6, 7, 9, and 10) are homologous to the yeast HDA1 protein and are found in both the nucleus and cytoplasm; and class III HDACs form a structurally distinct class of NAD-dependent enzymes that are similar to the yeast SIR2 proteins. Since inappropriate silencing of critical genes can result in one or both hits of tumor suppressor gene (TSG) inactivation in cancer, theoretically the reactivation of affected TSGs could have an enormous therapeutic value in preventing and treating cancer. Indeed, several HDAC inhibitors are currently being developed and tested for their potency in cancer chemotherapy. Importantly, these agents are also potentially applicable to chemoprevention if their toxicity can be minimized. Despite the toxic side effects and lack of specificity of some of the inhibitors, progress is being made. With the elucidation of the structures, functions and modes of action of HDACs, finding agents that may be targeted to specific HDACs and potentially reactivate expression of only a defined set of affected genes in cancer will be more attainable. PMID- 12724215 TI - Genome-wide analysis of epigenetics in cancer. AB - Human cancers are caused by multiple mechanisms. Research in the last 30 years has firmly established the roles of a group of genes including oncogenes, tumor suppressor genes, and DNA repair genes in human cancers. The activation and inactivation of these cancer genes can be caused by genetic mutations or epigenetic alterations. The epigenetic changes in cancers include methylation of CpG islands, loss of imprinting, and chromatin modification. The completion of the genome sequences of many organisms including the human has transformed the traditional approach to molecular biology research into an era of functional genome research. Traditional research usually involves the study of one or a few genes (proteins) in a particular biological process in normal physiology or disease. Functional genome research takes advantage of newly available genome sequences and high-throughput genome technologies to study genes and/or proteins to inform the perspective of entire biological processes. I will focus on recent progress in the identification of imprinted genes and methylation of CpG islands through genome-wide analysis. PMID- 12724216 TI - The development of CpG island methylation biomarkers using restriction landmark genomic scanning. AB - CpG island hypermethylation is a common occurrence in cancer. Because this is a stable molecular alteration of the DNA, which can be detected easily from very small amounts, DNA methylation is an attractive candidate to use as a molecular biomarker. Recent studies have used DNA methylation of genes known to be targets of genetic disruption in cancer as biomarkers for early detection of cancer, classification of malignancies, response to drug treatment, and as markers predictive of outcome. Since many of the currently used targets of methylation are methylated at rather low frequencies in various cancer types even though the gene may be frequently disrupted by other mechanisms, it would be useful to develop additional markers that are methylated at high frequency in the cancer being studied. Restriction landmark genomic scanning has been used for the identification of frequent targets of methylation in multiple malignancies. These markers, which can be either cancer type-specific or nonspecific, may prove to be effective biomarkers for diagnostic or prognostic purposes, or for midpoint analysis of intervention strategies. PMID- 12724217 TI - Sensitive detection of DNA methylation. AB - In recent years, many molecular biomarkers have been discovered that are capable of distinguishing tumors from normal tissue. Among the different types of markers, DNA methylation markers stand out for their potential to provide a unique combination of specificity, sensitivity, high information content, and applicability to a wide variety of clinical specimens. Methylation markers are particularly suited for situations where sensitive detection is necessary, such as when tumor DNA is either scarce or diluted by excess normal DNA. One of the most widely used methods for measuring methylation levels, methylation-specific PCR (MSP), has been proved to be a very effective tool in situations requiring sensitive detection. The addition of fluorogenic probes makes these assays more informative, quantitative, and suitable for a clinical format. The field of sensitive detection is not limited to MSP; hence, an alternative methylation sensitive amplification is discussed. PCR-based methylation assays have been applied to the detection of tumor DNA in a variety of body fluids, including serum, plasma, urine, sputum, and lavage fluids. In many cases, the sensitivity and specificity of these detection assays has been impressive, but important technological issues remain in areas such as sample preparation, assay design, and marker selection. Once these technical concerns have been addressed, the sensitive detection of methylation will provide a powerful diagnostic and prognostic tool, especially for the early detection of preneoplastic and neoplastic lesions. PMID- 12724218 TI - Methylation-sensitive representational difference analysis and its application to cancer research. AB - Methylation-sensitive representational difference analysis (MS-RDA) was previously established to detect differences in the methylation status of two genomes. This method uses the digestion of genomic DNA with a methylation sensitive restriction enzyme, HpaII, and PCR to prepare "HpaII-amplicons," followed by RDA. An HpaII-amplicon prepared using betaine and reverse electrophoresis was enriched 3.6-fold (compared with the HpaII-amplicon prepared by the original method) with DNA fragments originating from CpG islands (CGIs). As for the specificity of MS-RDA, it was shown that DNA fragments that are unmethylated in the tester and almost completely methylated in the driver are efficiently isolated. This indicated that genes that are in biallelic methylation or in monoallelic methylation with loss of the other allele are efficiently isolated. Further, by use of two additional methylation-sensitive six-base recognition restriction enzymes, SacII and NarI, more DNA fragments were isolated from CGIs in the 5' regions of genes. After analysis of human lung, gastric, and breast cancers, 12 genes were seen to be silenced and additional genes seen to show decreased expression in association with methylation of genomic regions outside CGIs in the 5' regions of genes. MS-RDA is effective in identifying silenced genes in various cancers. PMID- 12724219 TI - Cancer biomarker validation: standards and process: roles for the National Institute of Standards and Technology (NIST). AB - Rigorous validation of biomarkers for early detection of cancer differs at the National Institute of Standards and Technology (NIST) from similar processes common among research laboratories. As a newly discovered biomarker assay makes the transition from a research setting to the clinical diagnostic laboratory, it should progress through defined stages of assay confirmation. The first task of a validation laboratory is evaluation of research assay technology, performance, and specifications (analytical validation). However, the ultimate goal is initial validation of the test to identify early stage cancer (clinical validation). Upon technical and clinical confirmation, assays are moved systematically toward a standardized, reproducible, high-throughput format for clinical diagnostic implementation. With laboratory performance rigorously established, the clinical variables can subsequently be analyzed to define limitations, applications, and clinical utility. The role of NIST in technology evaluation for early cancer testing is described in the context of similar programs and prior experience at NIST. Here we conceptualize the validation steps of cancer test development and examine how NIST activities impact health care through institutional focus on measurement, technology, and standards development programs. PMID- 12724220 TI - Epigenetics and the environment. AB - DNA methylation and histone modification promote changes in chromatin structure that may affect gene expression in a heritable manner without directly altering the genome. As such, these phenomena are considered to be epigenetic in nature and are believed to contribute to the normal processes of human development but also to aberrant disease states such as cancer. Epigenetic processes probably contribute mechanistically to toxicant-induced changes in gene expression and cancer. Nickel is a potent human carcinogen that has been shown to alter DNA methylation patterns and affect histone acetylation status. Both of these changes are associated with the proximity of the affected regions to heterochromatin. The two processes probably occur in concert in mammalian cells. However, in yeast cells, DNA methylation is absent, and nickel is capable of regulating gene expression through changes in acetylation of the lysine residues in the N terminal tail of histone H4. Arsenic is another important environmental carcinogen, and it is methylated during its metabolism. Hence, it has been proposed that arsenic metabolism may deplete intracellular methyl group stores and thereby lead to changes in DNA methylation that may be involved in carcinogenesis. However, the data concerning DNA methylation changes following arsenic exposure are equivocal, leading researchers to propose that DNA hypo- and hypermethylation are both important in the development of arsenic-induced cancers. Heightened awareness by toxicologists of the importance of epigenetics in normal human development and in carcinogenesis should lead to the identification of other toxicants that manifest their effects, at least in part, via epigenetic mechanisms. PMID- 12724221 TI - Environmental exposure, DNA methylation, and gene regulation: lessons from diethylstilbesterol-induced cancers. AB - DNA methylation is an epigenetic mechanism that regulates chromosomal stability and gene expression. Abnormal DNA methylation patterns have been observed in many types of human tumors, including those of the breast, prostate, colon, thyroid, stomach, uterus, and cervix. We and others have shown that exposure to a wide variety of xenobiotics during critical periods of mammalian development can persistently alter the pattern of DNA methylation, resulting in potentially adverse biological effects such as aberrant gene expression. Thus, this epigenetic mechanism may underlie the observed increased risk in adulthood of several chronic diseases, including cancer, in response to xenobiotic exposures early in life. We present here the lessons learned from studies on the effects of perinatal diethylstilbesterol (DES) exposure on the methylation pattern of the promoters of several estrogen-responsive genes associated with the development of reproductive organs. Perinatal DES exposure, which induces epithelial tumors of the uterus in mice and is associated with several reproductive tract abnormalities and increased vaginal and cervical cancer risk in women, provides a clear example of how estrogenic xenobiotic exposure during a critical period of development can abnormally demethylate DNA sequences during organ development and possibly increase cancer risk later in life. In addition, nutritional factors and stress may also alter DNA methylation during early life and modulate the risk of cancer and other chronic diseases in adulthood. We suggest that DNA methylation status may be influenced by environmental exposures in early life, leading to increased risk of cancer in adulthood. PMID- 12724222 TI - Viral genes and methylation. AB - Epigenetics represents a new frontier in cancer research. Methylation is the best studied of the epigenetic mechanisms that regulate gene expression. Regulation of gene expression by means of methylation has been reported for tumor suppressor genes, oncogenes, viral promoters, and age-related genes. In this review, the regulation of viral gene expression by methylation is discussed, with particular emphasis on: (1) the virus-specific factors that bind to promoter regions; (2) the implications of this knowledge for designing viral vectors that can be used to deliver genes for the purpose of gene therapy; and (3) the use of this knowledge for the early detection and prevention of cancer. Since methylation can be reversed by a variety of exogenous agents, great potential exists to develop interventions that target cancer-associated aberrant methylation in an effort to reverse or prevent carcinogenesis. PMID- 12724223 TI - Epidemiologic considerations to assess altered DNA methylation from environmental exposures in cancer. AB - Epidemiologic studies in human populations have identified a broad spectrum of risk factors for cancer. Gene-damaging agents have been a primary focus of cancer epidemiology; however, all xenobiotics do not interact with DNA directly. Some exogenous agents induce epigenetic changes. In view of this, markers that measure changes to the epigenome must also be incorporated into molecular epidemiologic studies. We review the current understanding of the impact of exogenous agents including: micronutrients, chemotherapeutic agents, metals, and others, on DNA methylation. Two categories of genes are described: (1) genes that can alter susceptibility to aberrant DNA methylation and (2) genes that increase susceptibility to cancer when they are silenced through DNA methylation. Methods for incorporating markers of DNA methylation status into etiologic investigations of the impact of environmental exposures on disease (e.g., cancer) are discussed. PMID- 12724224 TI - Diet and DNA methylation interactions in cancer prevention. AB - Epigenetic events constitute an important mechanism by which gene function is selectively activated or inactivated. Since epigenetic events are susceptible to change they offer potential explanations of how environmental factors, including diet, may modify cancer risk and tumor behavior. Abnormal methylation patterns are a nearly universal finding in cancer, as changes in DNA methylation have been observed in many cancer tissues (e.g., colon, stomach, uterine cervix, prostate, thyroid, and breast). Site-specific alterations in DNA methylation have also been observed in cancer and may play a significant role in gene regulation and cancer development. This review presents intriguing evidence that part of the anticancer properties attributed to several bioactive food components, encompassing both essential nutrients and non-essential components, may relate to DNA methylation patterns. Four sites where dietary factors may be interrelated with DNA methylation are discussed. First, dietary factors may influence the supply of methyl groups available for the formation of S-adenosylmethionine (SAM). Second, dietary factors may modify the utilization of methyl groups by processes including shifts in DNA methyltransferase (Dnmt) activity. A third plausible mechanism may relate to DNA demethylation activity. Finally, the DNA methylation patterns may influence the response to a bioactive food component. PMID- 12724225 TI - Promoter hypermethylation as an epigenetic component in Type I and Type II endometrial cancers. AB - Epigenetic mechanisms that result in aberrant gene expression are a prominent feature of many cancer types. One main epigenetic mechanism for gene silencing involves promoter hypermethylation. Type I and type II endometrial cancers exhibit differing clinical, histologic, and molecular genetic characteristics. We hypothesize that these differences also extend to epigenetic phenomena. Promoter methylation analysis of 11 genes in a panel of endometrial cancers supports this hypothesis. These initial data indicate that promoter hypermethylation events occur frequently in type 1 cancer and were not detected in type II cancers using this panel of loci. These data tend to support the hypothesis that type I and type II endometrial cancers will exhibit distinct patterns of gene silencing based on promoter hypermethylation events. PMID- 12724226 TI - Epigenetics in carcinogenesis and cancer prevention. AB - Carcinogenesis is a stepwise process of accumulation of genetic and epigenetic abnormalities that can lead to cellular dysfunction. It has become clear that epigenetic changes are equally important for this multistep process to produce its results. This article describes the different roles that epigenetic modulation may play during carcinogenesis and how an early detection and chemopreventive intervention strategy that takes both sides of the equation into account would be advantageous. PMID- 12724227 TI - Histone acetylation and gastrointestinal carcinogenesis. AB - The importance of altered histone acetylation in gastrointestinal carcinogenesis, especially in relation to invasion and metastasis, is described. Histone acetylation and chromatin remodeling linked with CpG island methylation play a major role in epigenetic regulation of gene expression. Acetylation of histones through an imbalance of histone acetyltransferases and deacetylases disrupts nucleosome structure, which leads to DNA relaxation and subsequent increase in accessibility to transcription factors. The expression of acetylated histone H4 is reduced in a majority of gastric and colorectal cancers, indicating the low level of global histone acetylation in tumor cells. Moreover, reduced histone acetylation is significantly associated with depth of tumor invasion and nodal metastasis of gastrointestinal cancers. A histone deacetylase inhibitor, trichostatin A (TSA), induces growth arrest and apoptosis and suppresses invasion of cancer cells. Treatment with TSA, which is followed by increased histone acetylation in the promoters, induces the expression of many genes that are suppressors of invasion and metastasis, including tissue inhibitors of metalloproteinase and nm23H1/H2, in addition to negative cell cycle regulators and apoptosis-related molecules. Our approach, serial analysis of gene expression (SAGE), enabled us to identify a gene that is a novel candidate for a metastasis suppressor, whose expression is induced by histone acetylation. These findings suggest that, by modifying gene expression, histone deacetylation may participate not only in tumorigenesis but also in invasion and metastasis. Therefore, histone acetylation should be a promising target for cancer therapy, especially against invasive and metastatic disease, but also for cancer prevention. PMID- 12724228 TI - Epigenetics in high-grade astrocytomas: opportunities for prevention and detection of brain tumors. AB - Human high-grade astrocytomas (HGA) are the most prevalent incurable brain tumors. We found that the vast majority of HGA patients overexpress a restricted receptor for an immune regulatory cytokine, interleukin 13 (IL-13). Interestingly, the HGA-associated restricted receptor protein IL-13Ralpha2 is expressed in the testes, and its gene is localized to chromosome X. These mirror the expression pattern and genomic localization of cancer/testes tumor antigens (CTA). Hypothetical considerations and now experimental evidence are beginning to point towards epigenetics, and DNA methylation alterations in particular, as being responsible for the appearance in cancer of CTA, including IL-13Ralpha2. In line with our interest in the X chromosome and oncogenesis, we have identified a new ubiquitous angiogenic factor in HGA, a vascular endothelial growth factor-D (VEGF-D). We have also demonstrated that the activating protein-1 (AP-1) family of transcription factors play a potentially critical role in the progression of gliomas by eliciting uncontrolled upregulation of VEGF-D and other compounds essential for cancer cell proliferation, tumorigenesis, and infiltration. The possibility exists that an unopposed constitutive increase in AP-1 activity in HGA is related to epigenetic silencing of the inhibitors of AP-1 activity. These phenomena offer potential targets for exploitation in either prevention or early detection of brain tumors. For example, anticancer vaccines against shared CTA could help in prevention of HGA development. Furthermore, drugs with anti-AP-1 activity could be effective in preventing formation/progression of HGA, or progression from less malignant lower grade gliomas to HGA. Also, circulating antibodies against CTA and factors that are AP-1 regulated may provide a useful tool in early detection of brain tumors or for monitoring their progression following initial treatment. PMID- 12724229 TI - Aberrant DNA methylation in ovarian cancer: is there an epigenetic predisposition to drug response? AB - Epigenetic regulation of gene expression has been observed in a variety of tumor types. We have used microarray technology to evaluate the predisposition of drug response by aberrant methylation in ovarian cancer. Results indicate that loss of gene activity due to hypermethylation potentially confers a predisposition in certain cancer types and is an early event in disease progression. Methylation profiles of ovarian cancer might be useful for early cancer detection and prediction of chemotherapy outcome in a clinical context. PMID- 12724230 TI - DNA methylation, a biomarker for colorectal cancer: implications for screening and pathological utility. AB - Currently up to one-third of colorectal cancer patients present with locally advanced or metastatic disease that precludes a surgical cure. Performance limitations and low uptake of current screening tools have fueled research to develop minimally invasive approaches that can detect early-stage neoplasms. The observation that altered DNA can be amplified from the stool or circulation has stimulated research on its use as a biomarker of occult neoplasia. De novo methylation of CpG islands 5' to certain tumor suppressor genes has been associated with epigenetic silencing. At certain loci this phenomenon is specific for neoplastic populations, and it is frequently detected at early stages in colorectal tumorigenesis. Accordingly, hypermethylation events have been proposed by researchers as ideal targets for the basis of a screening panel to detect peripheral tumor DNA. This critique reviews research findings on the use of epigenetic biomarkers in screening for occult neoplasia. In addition, the authors consider the pathological utility of epigenetic testing in refining tumor staging and predicting disease recurrence. PMID- 12724231 TI - Epigenetic regulation of ARHI in breast and ovarian cancer cells. AB - ARHI (Ras homologue member I) encodes a 26-kDa GTPase with 50-60% amino acid homology to Ras and Rap. ARHI and Ras share similar GTP/GDP binding domains, but exert opposite functions. ARHI is one of the first reported tumor suppressors in the ras superfamily. ARHI is expressed consistently in normal breast and ovarian epithelial cells, but not in breast or ovarian cancers. The loss of ARHI can be related to tumor progression. Reexpression of ARHI induces apoptosis of breast and ovarian cancer cells by a caspase-independent, calpain-dependent pathway. ARHI is consistently expressed in normal breast and ovarian epithelial cells but is dramatically downregulated in more then 70% of breast and ovarian cancers. ARHI is maternally imprinted with methylation of the three CpG islands in the maternal allele of normal cells. ARHI is expressed only from the paternal allele whose three CpG islands are not methylated. Loss of ARHI expression can occur through a genetic event, with loss of heterozygosity observed in 40% of breast, ovarian, and pancreatic cancers; but it can also occur through epigenetic mechanisms, including DNA methylation, histone deacetylation, histone methylation, and transcriptional regulation. Our data suggest that acetylation and methylation of chromatin associated with the ARHI promoter leads to loss of both ARHI expression and the ability to suppress tumor growth. Changes in chromatin that silence ARHI may be driven by methylation-dependent and independent pathways. Reactivation of both the silenced paternal and imprinted maternal alleles can be achieved by demethylation and inhibition of histone deacetylation. PMID- 12724233 TI - DNA methylation as a cancer-specific biomarker: from molecules to populations. AB - Cancer contributes to a large proportion of the mortality and morbidity in the United States and worldwide. Despite advances in diagnosis and treatment of various cancers, early detection and treatment of cancer remain a challenge. Diagnosis of cancer often occurs once the disease has progressed to a point where currently available intervention options provide limited success. Therefore, techniques that enable early detection followed by targeted interventions would influence stage at diagnosis and, in turn, mortality associated with cancer. Identification of molecular biomarkers, especially those that are associated with cancer initiation and progression, shows promise as an effective strategy in this regard. One potential early detection biomarker is DNA methylation of the promoter region of certain cancer-associated genes, which results in gene inactivation. Examination of serum for circulating tumor DNA with abnormal methylation patterns offers a possible method for early detection of several cancers and serves as a point for early intervention and prevention strategies. Additionally, it is imperative to consider how such a screening mechanism can be implemented in populations at risk, especially in resource-poor settings. Thus, the challenge is to validate DNA methylation as a cancer-specific biomarker, with the ultimate goal of designing a research plan that integrates the current knowledge base regarding cancer detection and diagnosis into specific prevention and intervention strategies that can be applied at a population level. PMID- 12724232 TI - Epigenetic changes: potential therapeutic targets. AB - Recent advances in human genome research have resulted in novel approaches for the identification of epigenetic modifications associated with cancer. Modulators of DNA methylation and chromatin structure have a dramatic effect on gene expression, cellular proliferation, differentiation, and apoptosis. Molecular pathways regulating epigenetic events that occur during tumorigenesis have been exploited as new targets for therapeutic intervention. Clinical studies exploring the effectiveness of therapeutic agents targeting DNA methylation and acetylation of histones have yielded promising results. Molecular profiles of epigenetic alterations in cancer cells could allow better stratification of patients who may show responsiveness to specific treatments. PMID- 12724236 TI - Pathways to emotional dysfunction in first-episode psychosis. PMID- 12724237 TI - Validity issues in transcultural epidemiology. PMID- 12724234 TI - Early detection and risk assessment: proceedings and recommendations from the Workshop on Epigenetics in Cancer Prevention. AB - Recent advances in molecular biology that have provided a greater understanding of multistage carcinogenesis include the use of biomarkers of early detection and risk assessment. Prominent among such biomarkers are epigenetic changes. The field of epigenetics has seen a recent surge of interest among cancer researchers since alterations in DNA methylation have emerged as one of the most consistent molecular alterations in multiple neoplasms. Chromatin condensation, histone deacetylation, and promoter methylation are major steps in the epigenetic regulation of gene expression. Epigenetic changes may occur due to environmental factors, aging, and genomic imprinting. An important distinction between genetic and epigenetic alterations in cancer prevention is that the latter might be more easily reversed using therapeutic interventions. In the workshop the following areas of research were recognized for emphasis in future work: (1) basic epigenetic mechanisms in cancer need further investigation; (2) technology development in the area of epigenetics, such as high-throughput quantitative assays and increased sensitivity/specificity, is essential for the early detection and risk assessment of cancer; (3) the clinical application of epigenetic changes to cancer prevention and risk assessment needs further investigation. Further research will lead to the identification of new targets for cancer prevention. PMID- 12724238 TI - Sudden infant death syndrome and psychiatric disorders. PMID- 12724239 TI - The eye's mind: brain mapping and psychiatry. PMID- 12724240 TI - Should psychiatrists read fiction? PMID- 12724241 TI - Is academic psychiatry for sale? PMID- 12724242 TI - Psychiatric disorder in veterans of the Persian Gulf War of 1991. Systematic review. AB - BACKGROUND: Veterans of the Persian Gulf War of 1991 have reported symptoms attributed to their military service. AIMS: To review all studies comparing the prevalence of psychiatric disorders in Gulf War veterans and in a comparison group of service personnel not deployed to the Gulf War. METHOD: Studies of military personnel deployed to the Gulf published between 1990 and 2001 were identified from electronic databases. Reference lists and websites were searched and key researchers were contacted for information. A total of 2296 abstracts and 409 complete articles were reviewed and data were extracted independently by two members of the research team. RESULTS: The prevalence of psychiatric disorder in 20 studies of Gulf War veterans was compared with the prevalence in the comparison group. Prevalence of post-traumatic stress disorder (PTSD) and common mental disorder were higher in the Gulf War veterans. Heterogeneity between studies was significant, but all reported this increased prevalence. CONCLUSIONS: Veterans of the Persian Gulf War reported an increased prevalence of PTSD and common mental disorder compared with other active service personnel not deployed to the Gulf. These findings are attributable to the increase in psychologically traumatic events in wartime. PMID- 12724243 TI - Status of neurosurgery for mental disorder in Scotland. Selective literature review and overview of current clinical activity. AB - BACKGROUND: Despite the application of ablative neurosurgical treatments for intractable mental disorder throughout most of the past century, unequivocal evidence for efficacy has not been provided. AIMS: To review the status of ablative neurosurgery for mental disorder and to describe the activities of the Scottish national service. METHOD: Relevant literature is reviewed alongside a description of recent clinical activity. RESULTS: Neurosurgical treatment is offered to a small number of patients severely disabled by otherwise intractable mental disorder. There are inequalities in the strength of evidence to support the use of some of these procedures. The frequency and severity of adverse effects remains unclear. We are collecting data that should inform future practice. CONCLUSIONS: Modern neurosurgery can offer clinically meaningful symptom relief and improved function for 'untreatable' patients with chronic, severe depression and obsessive-compulsive disorder. However, follow-up studies of greater rigour are required. The potential role of non-ablative alternatives remains unclear. PMID- 12724244 TI - Controlled trial of the short- and long-term effect of psychological treatment of post-partum depression. I. Impact on maternal mood. AB - BACKGROUND: Psychological interventions for postnatal depression can be beneficial in the short term but their longer-term impact is unknown. AIMS: To evaluate the long-term effect on maternal mood of three psychological treatments in relation to routine primary care. METHOD: Women with post-partum depression (n=193) were assigned randomly to one of four conditions: routine primary care, non-directive counselling, cognitive-behavioural therapy or psychodynamic therapy. They were assessed immediately after the treatment phase (at 4.5 months) and at 9, 18 and 60 months post-partum. RESULTS: Compared with the control, all three treatments had a significant impact at 4.5 months on maternal mood (Edinburgh Postnatal Depression Scale, EPDS). Only psychodynamic therapy produced a rate of reduction in depression (Structured Clinical Interview for DSM-III - R) significantly superior to that of the control. The benefit of treatment was no longer apparent by 9 months post-partum. Treatment did not reduce subsequent episodes of post-partum depression. CONCLUSIONS: Psychological intervention for post-partum depression improves maternal mood (EPDS) in the short term. However, this benefit is not superior to spontaneous remission in the long term. PMID- 12724245 TI - Controlled trial of the short- and long-term effect of psychological treatment of post-partum depression: 2. Impact on the mother-child relationship and child outcome. AB - BACKGROUND: Postnatal depression is associated with adverse child cognitive and socio-emotional outcome. It is not known whether psychological treatment affects the quality of the mother-child relationship and child outcome. AIMS: To evaluate the effect of three psychological treatments on the mother-child relationship and child outcome. METHOD: Women with post-partum depression (n=193) were assigned randomly to routine primary care, non-directive counselling, cognitive behavioural therapy or psychodynamic therapy. The women and their children were assessed at 4.5, 18 and 60 months post-partum. RESULTS: Indications of a positive benefit were limited. All three treatments had a significant benefit on maternal reports of early difficulties in relationships with the infants; counselling gave better infant emotional and behaviour ratings at 18 months and more sensitive early mother-infant interactions. The treatments had no significant impact on maternal management of early infant behaviour problems, security of infant-mother attachment, infant cognitive development or any child outcome at 5 years. CONCLUSIONS: Early intervention was of short-term benefit to the mother-child relationship and infant behaviour problems. More-prolonged intervention may be needed. Health visitors could deliver this. PMID- 12724246 TI - Depersonalisation disorder: clinical features of 204 cases. AB - BACKGROUND: Depersonalisation disorder is a poorly understood and underresearched syndrome. AIMS: To carry out a large and comprehensive clinical and psychopathological survey of a series of patients who made contact with a research clinic. METHOD: A total of 204 consecutive eligible referrals were included: 124 had a full psychiatric examination using items of the Present State Examination to define depersonalisation/derealisation and 80 had either a telephone interview (n=22) or filled out a number of self-report questionnaires. Cases assessed were diagnosed according to DSM-IV criteria. RESULTS: The mean age of onset was 22.8 years; early onset was associated with greater severity. There was a slight male preponderance. The disorder tended to be chronic and persistent. Seventy-one per cent met DSM-IV criteria for primary depersonalisation disorder. Depersonalisation symptom scores correlated with both anxiety and depression and a past history of these disorders was commonly reported. 'Dissociative amnesia' was not prominent. CONCLUSIONS: Depersonalisation disorder is a recognisable clinical entity but appears to have significant comorbidity with anxiety and depression. Research into its aetiology and treatment is warranted. PMID- 12724247 TI - Seasonality, negative life events and social support in a community sample. AB - BACKGROUND: Seasonal affective disorder (SAD) is now a well-described form of depressive disorder. However, relatively little research has focused upon psychosocial factors and SAD. AIMS: To determine the association between demographic/psychosocial factors and increased reported seasonal patterns of mood disorder (seasonality) and SAD in a community sample in the UK. METHOD: A total of 1250 people, aged between 18 and 64 years, randomly selected from a primary care database were screened for SAD. Those above cut-off underwent diagnostic interview and completed several self-report questionnaires. Multivariate analysis was conducted to determine which variables were significantly associated with increased seasonality. RESULTS: Four factors (having experienced more numerous negative life events, having low levels of social support, being a woman and being non-native) were predictive of higher seasonality. Being a woman was predictive of being diagnosed as a case of SAD. CONCLUSIONS: A new association has been identified between increased seasonality, negative life events and social support. Future research should assess the psychosocial causes or consequences of SAD while continuing to examine the biology of the condition. PMID- 12724248 TI - Structural disconnectivity in schizophrenia: a diffusion tensor magnetic resonance imaging study. AB - BACKGROUND: There is growing evidence that schizophrenia is a disorder of cortical connectivity. Specifically, frontotemporal and frontoparietal connections are thought to be functionally impaired. Diffusion tensor magnetic resonance imaging (DT-MRI) is a technique that has the potential to demonstrate structural disconnectivity in schizophrenia. AIMS: To investigate the structural integrity of frontotemporal and frontoparietal white matter tracts in schizophrenia. METHOD: Thirty patients with DSM-IV schizophrenia and thirty matched control subjects underwent DT-MRI and structural MRI. Fractional anisotropy - an index of the integrity of white matter tracts - was determined in the uncinate fasciculus, the anterior cingulum and the arcuate fasciculus and analysed using voxel-based morphometry. RESULTS: There was reduced fractional anisotropy in the left uncinate fasciculus and left arcuate fasciculus in patients with schizophrenia compared with controls. CONCLUSIONS: The findings of reduced white matter tract integrity in the left uncinate fasciculus and left arcuate fasciculus suggest that there is frontotemporal and frontoparietal structural disconnectivity in schizophrenia. PMID- 12724249 TI - Mental health of Kosovan Albanian refugees in the UK. AB - BACKGROUND: In 1999 the UK received 4346 refugees from Kosovo. AIMS: To determine the prevalence of mental health problems in this group. METHOD: A sample of 842 adults was surveyed. All were asked to complete self-report questionnaires (translated into Kosovan Albanian). A subset of 120 participants were later interviewed in Albanian using the Clinician Administered PTSD (post-traumatic stress disorder) Scale and a depression interview. RESULTS: The study yielded estimates of prevalence of PTSD and depression. Self-report measures appear to overestimate the prevalence of these disorders. Just under half of the group surveyed had a diagnosis of PTSD and less than one-fifth had a major depressive disorder. CONCLUSIONS: These results may be taken as a sign of the resilience of many who survived this conflict but they also imply that there is still a substantial need for good health and social care in a significant proportion. Psychosocial interventions are likely to be an important part of the treatment programme. PMID- 12724250 TI - Mild cognitive impairment: prevalence and incidence according to different diagnostic criteria. Results of the Leipzig Longitudinal Study of the Aged (LEILA75+). AB - BACKGROUND: Although mild cognitive impairment is associated with an increased risk of developing dementia, there has been little work on its incidence and prevalence. AIMS: To report age-specific prevalence, incidence and predictive validities for four diagnostic concepts of mild cognitive impairment. METHOD: A community sample of 1045 dementia-free individuals aged 75 years and over was examined by neuropsychological testing in a three-wave longitudinal study. RESULTS: Prevalence rates ranged from 3% to 20%, depending on the concept applied. The annual incidence rates applying different case definitions varied from 8 to 77 per 1000 person-years. Rates of conversion to dementia over 2.6 years ranged from 23% to 47%. CONCLUSIONS: Mild cognitive impairment is frequent in older people. Prevalence, incidence and predictive validities are highly dependent on the diagnostic criteria applied. PMID- 12724251 TI - Betel use and schizophrenia. PMID- 12724252 TI - ECT and old age psychiatry. PMID- 12724253 TI - Problem substance use and schizophrenia. PMID- 12724254 TI - Lithium augmentation in treatment-refractory unipolar depression. PMID- 12724255 TI - Getting closer to suicide prevention. PMID- 12724256 TI - Vulnerable individuals and the Human Rights Act. PMID- 12724257 TI - Concepts of social capital. PMID- 12724259 TI - IGDA. Introduction. PMID- 12724260 TI - IGDA. 1: Conceptual bases--historical, cultural and clinical perspectives. PMID- 12724261 TI - IGDA. 2: Interviewing the patient. PMID- 12724262 TI - IGDA. 3: Use of extended sources of information. PMID- 12724263 TI - IGDA. 4: Evaluation of symptoms and mental state. PMID- 12724265 TI - IGDA. 6: Supplementary assessment procedures--functioning, social context, cultural framework and quality of life. PMID- 12724264 TI - IGDA.5: Supplementary assessment procedures--psychopathological, neuropsychological and physical aspects. PMID- 12724266 TI - IGDA. 7: Standardised multi-axial diagnostic formulation. PMID- 12724267 TI - IGDA. 8: Idiographic (personalised) diagnostic formulation. PMID- 12724268 TI - IGDA. 9: Linking diagnosis to care--treatment planning. PMID- 12724269 TI - IGDA. 10: Organising the clinical chart. PMID- 12724270 TI - IGDA. 11: Illustrative clinical case. PMID- 12724271 TI - Oviduct cells express the cyclic AMP-adenosine pathway. AB - The extracellular cAMP-adenosine pathway refers to the local production of adenosine mediated by cAMP egress into the extracellular space, conversion of cAMP to AMP by ectophosphodiesterase (PDE), and the metabolism of AMP to adenosine by ecto-5'-nucleotidase. The goal of this study was to assess whether the cAMP-adenosine pathway is expressed in oviduct cells. Studies were conducted in cultured bovine oviduct cells (mixed cultures of fibroblasts and epithelial cells, 1:1 ratio). Confluent monolayers of oviduct cells were exposed to cAMP (0.01-100 micromol/L) in the presence and absence of 3-isobutyl-1-methylxanthine (IBMX, 1 mmol/L, an inhibitor of both extracellular and intracellular PDE activity), 1,3-dipropyl-8-p-sulfophenylxanthine (DPSPX, 100 micromol/L, a xanthine that can inhibit extracellular or ecto-PDE activity at high concentrations), or alpha,beta-methylene-adenosine-5'-diphosphate (AMPCP, 100 micromol/L, an ecto-5'-nucleotidase inhibitor) for 0-60 min. The medium was then sampled and assayed for AMP, adenosine, and inosine. Addition of exogenous cAMP to oviduct cells increased extracellular levels of AMP, adenosine, and inosine in a concentration- and time-dependent manner. This effect was attenuated by blockade of total (extracellular and intracellular) PDE activity (IBMX), ecto-PDE activity (DPSPX), or ecto-5'-nucleotidase (AMPCP). The functional relevance of the cAMP-adenosine pathway is supported by the findings that treatment with adenylyl cyclase stimulants (forskolin plus isoproterenol) resulted in the egress of cAMP (97% extracellular) into the extracellular space and its conversion into adenosine. The extracellular cAMP-adenosine pathway exists in oviduct cells and may play an important role in regulating the biology and physiology of the oviduct. This pathway also may play a critical role in regulating sperm function, fertilization, and early embryo development. PMID- 12724273 TI - Hormonal induction and stability of monosex populations in the medaka (Oryzias latipes): expression of sex-specific marker genes. AB - The model teleost medaka (Oryzias latipes, d-rR.YHNI strain) was used to produce offspring of a defined sex (monosex populations) by crossing experimentally produced YY and XX males to normal females. These monosex populations had the predicted chromosomal constitution as shown by a sex chromosome-specific DNA sequence. However, in XX populations the spontaneous development of males without previous exposure to androgens was observed. Differences in the percentage of male offspring from individual XX breeding pairs indicate a possible variation of unknown genetic factors to be responsible for the development of XX males. The expression of two gonadal genes that are involved in sex differentiation, Dmrt1b(Y) and Fig1a (factor in the germ line alpha), was analyzed in monosex populations. Dmrt1b(Y) expression correlated strictly with the genotype but not the sexual phenotype. When XY juvenile fish were exposed to 17 alpha ethynylestradiol at concentrations that induce sex reversal, Dmrt1b(Y) expression was not repressed. However, Dmrt1b(Y) was expressed in XY or YY gonads regardless of the sex and could not be detected in XX individuals. In contrast, the expression of Fig1a correlated with the phenotypic sex: Fig1a was expressed in male juvenile fish exposed to 17 alpha-ethynylestradiol and repressed in fish exposed to 17 alpha-methyltestosterone. The Dmrt1b(Y) expression appears to reflect an early and important event in sex determination and lends support to the suggested key regulatory role of the Dmrt1b(Y) gene in sex determination. This process is apparently hormone insensitive, and the expression of further downstream acting genes can be regulated (directly or indirectly) by sex steroids. PMID- 12724272 TI - Effects of prostaglandin F2alpha and progesterone on the ability of bovine luteal cells to stimulate T lymphocyte proliferation. AB - Bovine luteal cells express class I and II major histocompatibility complex molecules and stimulate T lymphocyte proliferation in vitro. Proliferation of T lymphocytes is greater in cocultures of luteal cells and T lymphocytes collected following administration of a luteolytic dose of prostaglandin (PG) F2alpha to the cow. Whether this results from changes in luteal cells that increase their ability to stimulate T lymphocyte proliferation or from changes in T lymphocytes that enhance their ability to respond to luteal cells is unclear. To determine which is the case, luteal cell-T lymphocyte cocultures were performed using luteal cells and T lymphocytes isolated from the same animals before and 8 h after administration of PGF2alpha. In the presence of T lymphocytes collected before PGF2alpha administration, luteal cells isolated after PGF2alpha were more potent stimulators of T lymphocyte proliferation than were luteal cells collected before PGF2alpha (P<0.05). The effect of progesterone on luteal cell-stimulated T lymphocyte proliferation was also evaluated. Proliferation of T lymphocytes was greater (P<0.05) in cultures containing the cytochrome P450 side-chain cleavage enzyme-inhibitor aminoglutethimide. Exogenous progesterone caused a dose dependent inhibition of luteal cell-stimulated T lymphocyte proliferation (P<0.05). Progesterone-receptor mRNA was undetectable in peripheral blood mononuclear cells collected before and after PGF2alpha administration, indicating that the effect of progesterone was not mediated via progesterone receptors in lymphocytes. These results imply that specific changes in luteal cells in response to PGF2alpha enhance the ability of these cells to stimulate T lymphocyte proliferation. These results also demonstrate that progesterone can suppress luteal cell-stimulated T lymphocyte proliferation. PMID- 12724274 TI - Luteogenesis in cyclic ewes: echotextural, histological, and functional correlates. AB - To date, it has not been possible to detect corpus luteum (CL) by ultrasonography, immediately following ovulation, in the ewe. Early CL detection is essential to be able to relate luteal outcome to the developmental pattern of the ovulated follicle and to confirm ovulation. Image analysis of the CL may be useful in providing a noninvasive picture of CL differentiation and function. The present study was designed to use high-resolution ultrasonography to monitor and to correlate the echotextural, histological, and functional attributes of the developing ovine CL from Days 1 to 3 after ovulation. Ten ewes underwent twice daily transrectal ultrasonography and blood sampling from the day of synchronized estrus. Ewes were ovariectomized at 12-24, 36-48, and 60-72 h after ovulation. Ovaries collected were scanned in a water bath before processing for histology. Ultrasonographic images of CL were analyzed for echotexture. Histological sections were analyzed for the percentage area of the CL occupied by blood clot or luteal tissue. Serum samples were analyzed for progesterone concentration. Numerical pixel value, heterogeneity, and percentage of the CL occupied by blood clot declined (P<0.05) from 12-24 to 60-72 h after ovulation. Luteal area and serum progesterone concentration increased (P<0.05) from 12-24 to 60-72 h. The results indicated that it was possible to visualize developing CL as early as 12 24 h after ovulation in the ewe. Echotexture of the CL was closely associated with its morphological and functional characteristics; image analysis holds promise for noninvasive monitoring of CL differentiation and growth. PMID- 12724275 TI - Influence of porcine spermadhesins on the susceptibility of boar spermatozoa to high dilution. AB - The effect of heparin-binding and non-heparin-binding spermadhesins on the viability, motility, and mitochondrial activity of boar spermatozoa at the high dilution (300,000 sperm/ml) to which sperm are exposed during the process of sex sorting by flow cytometry was investigated. Incubation of spermatozoa with heparin-binding spermadhesins caused a time- and dose-dependent decrease in the percentage of functional spermatozoa. The percentage of viable spermatozoa incubated at 38 degrees C with heparin-binding spermadhesins diluted in PBS (1 mg/ml) dropped from 75% (0.5 h) to 4% (5 h), whereas the percentage of viable spermatozoa incubated in PBS without proteins (control) decreased from 85% (0.5 h) to 19% (5 h). Addition of non-heparin-binding PSP-I/PSP-II spermadhesin to the PBS resulted in a concentration-dependent increment of the percentage of viable cells (65% after 5-h incubation), with maximum effect at 1.5 mg/ml. The heparin binding spermadhesins totally suppressed sperm motility and mitochondrial activity after 5 h of incubation. The same parameters of sperm incubated in the presence of 1.5 mg/ml of PSP-I/PSP-II were 50% and 58%, respectively, and the percentages of control sperm displaying motility and mitochondrial activity were 21% and 26%, respectively. Moreover, the viability, motility, and mitochondrial activity all decreased on incubation of spermatozoa with mixtures of PSP-I/PSP-II and heparin-binding spermadhesins as the concentration of the latter increased. We conclude that PSP-I/PSP-II and the heparin-binding spermadhesins exert antagonistic effects on the functionality of highly diluted boar spermatozoa. The finding that PSP-I/PSP-II contributes to maintaining sperm with high viability, motility, and mitochondrial activity for at least 5 h at physiological temperature points to its potential use as an additive for sperm preservation, specifically of highly diluted, flow-sorted spermatozoa for sex preselection. PMID- 12724276 TI - Specific expression of VCY2 in human male germ cells and its involvement in the pathogenesis of male infertility. AB - Abnormal spermatogenesis in men with Y-chromosome microdeletions suggests that genes important for spermatogenesis have been removed from these individuals. VCY2 is a testis-specific gene that locates in the most frequently deleted azoospermia factor c region in the Y chromosome. We have raised an antiserum to VCY2 and used it to characterize the localization of VCY2 in human testis. Using Western blot analysis, the affinity-purified polyclonal VCY2 antibody gave a single specific band of approximately 14 kDa in size, corresponding to the expected size of VCY2 in all the collected human testicular biopsy specimens with normal spermatogenesis. Immunohistochemical analyses showed that VCY2 localized to the nuclei of spermatogonia, spermatocytes, and round spermatids, except elongated spermatids. At the ultrastructural level, VCY2 expression was found in the nucleus of human ejaculated spermatozoa. To determine the possible relationship of VCY2 with the pathogenesis of male infertility, we examined a group of infertile men with and without Y-chromosome microdeletions and with known testicular pathology using VCY2 antibody. VCY2 was weakly expressed at the spermatogonia and immunonegative in spermatocytes and round spermatids in testicular biopsy specimens with maturation arrest or hypospermatogenesis. The specific localization of the protein in germ cell nuclei indicates that VCY2 is likely to function in male germ cell development. The impaired expression of VCY2 in infertile men suggests its involvement in the pathogenesis of male infertility. PMID- 12724277 TI - A comparison of the frequency of sperm chromosome abnormalities in men with mild, moderate, and severe oligozoospermia. AB - Infertile men undergoing intracytoplasmic sperm injection have an increased frequency of chromosome abnormalities in their sperm. Men with low sperm concentration (oligozoospermia) have an increased risk of sperm chromosome abnormalities. This study was initiated to determine whether men with severe oligozoospermia (<10(6) sperm/ml) have a higher frequency of chromosome abnormalities in their sperm compared with men with moderate (1-9 x 10(6) sperm/ml) or mild (10-19 x 10(6) sperm/ml) oligozoospermia. Multicolor fluorescence in situ hybridization analysis was performed using DNA probes specific for chromosomes 13, 21, X, and Y (with chromosome 1 as an autosomal control for the sex chromosomes). Aneuploidy and disomy frequencies were assessed from a total of 603,011 sperm from 30 men: 10 in each of the categories. The mean frequencies of disomy for the patients with mild, moderate, and severe oligozoospermia were 0.17%, 0.24%, and 0.30%, respectively, for chromosome 13 and 0.22%, 0.44%, and 0.58%, respectively, for chromosome 21. For the sex chromosomes, the mean frequencies of disomy for mild, moderate, and severe oligozoospermia were 0.25%, 1.04%, and 0.68%, respectively, for XY, 0.047%, 0.08%, and 0.10%, respectively, for XX, and 0.04%, 0.06%, and 0.09%, respectively, for YY. The frequencies for diploidy also increased from 0.4% for mild to 1.20% for moderate to 1.24% for severe oligozoospermia. There was a significant inverse correlation between the frequency of sperm chromosome abnormalities and the sperm concentration for XY, XX, and YY disomy and diploidy. These results demonstrate that men with severe oligozoospermia have an elevated risk for chromosome abnormalities in their sperm, particularly sex chromosome abnormalities. PMID- 12724278 TI - Polyunsaturated fatty acids and bovine interferon-tau modify phorbol ester induced secretion of prostaglandin F2 alpha and expression of prostaglandin endoperoxide synthase-2 and phospholipase-A2 in bovine endometrial cells. AB - Embryonic mortality in cattle may occur because of inadequate inhibition of uterine secretion of prostaglandin (PG) F2alpha mediated by bovine interferon-tau (bIFN-tau). The objectives of the present study were to determine whether polyunsaturated fatty acids inhibit secretion of PGF2alpha from bovine endometrial cells induced by stimulating protein kinase C with phorbol 12,13 dibutyrate (PDBu) and to investigate possible mechanisms of action. Confluent cells were exposed for 24 h to 100 microM of linoleic, arachidonic (AA; C20:4, n 6), linolenic (LNA; C18:3, n-3), eicosapentaenoic (EPA; C20:5, n-3), or docosahexaenoic (DHA; C22:6, n-3) acid. After incubation, cells were washed and stimulated with PDBu. The EPA, DHA, and LNA attenuated secretion of PGF2alpha in response to PDBu. The EPA and DHA were more potent inhibitors than LNA. The EPA inhibited secretion of PGF2alpha at 6.25 microM. Secretion of PGF2alpha in response to PDBu decreased with increasing incubation time with EPA. Both bIFN tau and EPA inhibited secretion of PGF2alpha, and their inhibitory effects were additive. The bIFN-tau, but not EPA, reduced the abundance of PG endoperoxide synthase-2 (PGHS-2) mRNA. Incubation with 100 microM EPA, DHA, or AA for 24 h followed by treatment with PDBu did not affect concentrations of PGHS-2 and phospholipase A2 proteins. The EPA and DHA inhibit secretion of PGF2alpha through a mechanism different from that of bIFN-tau. The effect of EPA on PGF2alpha secretion may be caused by competition with AA for PGHS-2 activity or reduction of PGHS-2 activity. The use of EPA and DHA to inhibit uterine secretion of PGF2alpha and to improve embryonic survival in cattle warrants further investigation. PMID- 12724279 TI - Evidence for trichloroethylene bioactivation and adduct formation in the rat epididymis and efferent ducts. AB - Recent studies indicate that trichloroethylene (TCE) may be a male reproductive toxicant. It is metabolized by conjugation with glutathione and cytochrome p450 dependent oxidation. Reactive metabolites produced along both pathways are capable of forming protein adducts and are thought to be involved in TCE-induced liver and kidney damage. Similarly, in situ bioactivation of TCE and subsequent binding of metabolites may be one mechanism by which TCE acts as a reproductive toxicant. Cysteine-conjugate beta-lyase (beta-lyase) bioactivates the TCE metabolite dichlorovinyl cysteine (DCVC) to a reactive intermediate that is capable of binding cellular macromolecules. In the present study, Western blot analysis indicated that the soluble form of beta-lyase, but not the mitochondrial form, was present in the epididymis and efferent ducts. Both forms of beta-lyase were detected in the kidney. When rats were dosed with DCVC, no protein adducts were detected in the epididymis or efferent ducts, although adducts were present in the proximal tubule of the kidney. Trichloroethylene can also be metabolized and form protein adducts through a cytochrome p450-mediated pathway. Western blot analysis detected the presence of cytochrome p450 2E1 (CYP2E1) in the efferent ducts. Immunoreactive proteins were localized to efferent duct and corpus epididymis epithelia. Metabolism of TCE was demonstrated in vitro using microsomes prepared from untreated rats. Metabolism was inhibited 77% when efferent duct microsomes were preincubated with an antibody to CYP2E1. Dichloroacetyl adducts were detected in epididymal and efferent duct microsomes exposed in vitro to TCE. Results from the present study indicate that the cytochrome p450-dependent formation of reactive intermediates and the subsequent covalent binding of cellular proteins may be involved in the male reproductive toxicity of TCE. PMID- 12724280 TI - Thermoreversible gel formulation containing sodium lauryl sulfate as a potential contraceptive device. AB - The contraceptive properties of a gel formulation containing sodium lauryl sulfate were investigated in both in vitro and in vivo models. Results showed that sodium lauryl sulfate inhibited, in a concentration-dependent manner, the activity of sheep testicular hyaluronidase. Sodium lauryl sulfate also completely inhibited human sperm motility as evaluated by the 30-sec Sander-Cramer test. The acid-buffering capacity of gel formulations containing sodium lauryl sulfate increased with the molarity of the citrate buffers used for their preparations. Furthermore, experiments in which semen was mixed with undiluted gel formulations in different proportions confirmed their physiologically relevant buffering capacity. Intravaginal application of the gel formulation containing sodium lauryl sulfate to rabbits before their artificial insemination with freshly ejaculated semen completely prevented egg fertilization. The gel formulation containing sodium lauryl sulfate was fully compatible with nonlubricated latex condoms. Taken together, these results suggest that the gel formulation containing sodium lauryl sulfate could represent a potential candidate for use as a topical vaginal spermicidal formulation to provide fertility control in women. PMID- 12724281 TI - Physiological changes in oocyte-cumulus cell complexes from diabetic mice that potentially influence meiotic regulation. AB - We have previously shown that the type I diabetic condition significantly alters meiotic regulation in mouse oocytes. In the present study, possible physiological deficiencies underlying such meiotic dysfunction were examined in oocyte-cumulus cell complexes (OCC) from type I diabetic mice. Whereas the diabetic condition did not affect glycolysis or the tricarboxylic acid cycle, the increased flux of glucose through the pentose phosphate pathway in response to FSH treatment was suppressed. De novo purine synthesis was also compromised, and ATP levels were reduced in freshly isolated OCC. Additionally, diabetes resulted in a reduction in FSH-mediated cAMP synthesis. The responsiveness of the oocyte to cAMP was also affected; fewer oocytes were induced to resume maturation after a stimulatory pulse with cAMP analogs. Meiotic induction triggered by FSH was significantly reduced, but that stimulated by phorbol ester or epidermal growth factor was affected to a much lesser extent. In addition to metabolic deficiencies, the cell cell communication between the oocyte and the cumulus cells was reduced in diabetic mice as determined by coupling assays. Thus, numerous physiological parameters are affected by type I diabetes, and these changes may collectively contribute to altered meiotic regulation. PMID- 12724282 TI - Spatiotemporal changes of levels of a moonlighting protein, phospholipid hydroperoxide glutathione peroxidase, in subcellular compartments during spermatogenesis in the rat testis. AB - We studied temporal changes in the subcellular localization and levels of a moonlighting protein, phospholipid hydroperoxide glutathione peroxidase (PHGPx), in spermatogenic cells and mature sperm of the rat by immunofluorescence and immunoelectron microscopy. The PHGPx signals were detected in chromatoid bodies, clear nucleoplasm, mitochondria-associated material, mitochondrial aggregates, granulated bodies, and vesicles in residual bodies in addition to mitochondria, nuclei, and acrosomes as previously reported. Within mitochondria, PHGPx moved from the matrix to the outermost membrane region in step 19 spermatid, suggesting that this spatiotemporal change is synchronized with the functional change of PHGPx in mitochondria. In the nucleus, PHGPx was associated with electron-lucent spots and with the nuclear envelope, and PHGPx in the latter region increased after step 16. In early pachytene spermatids, PHGPx signals were noted in the nuclear material exhibiting a very similar density to chromatoid bodies and in the intermitochondrial cement, supporting the previous proposal that chromatoid bodies originate from the nucleus and intermitochondrial cement. The presence of PHGPx in such various compartments suggested versatile roles for this protein in spermatogenesis. Quantitative immunoelectron microscopic analysis also revealed dynamic changes in the labeling density of PHGPx in different subcellular compartments as follows: 1). Total cellular PHGPx rapidly increased after step 5 and reached a maximum at step 18; 2). mitochondrial labeling density increased after step 1 and achieved a maximum in steps 15-17; 3). nuclear labeling density suddenly increased in steps 12-14 to a maximum; 4). in cytoplasmic matrix, the density remained low in all steps; and 5). the labeling density in chromatoid bodies gradually decreased from pachytene spermatocytes to spermatids at step 18. These spatiotemporal changes in the level of PHGPx during the differentiation of spermatogenic cells to sperm infer that PHGPx plays a diverse and important biological role in spermatogenesis. PMID- 12724283 TI - Vasopressin receptor expression in the placenta. AB - The arginine vasopressin (AVP) type 1a receptor (V1a) is well known to mediate vasoconstriction. In pregnancy, blood flow in the placenta is crucial for sustaining normal growth and development of the fetus. This is the first AVP receptor study in the placenta and fetal membranes. The aim was to compare, quantitatively, the level of V1a gene expression with that of a known marker for vascularization, aquaporin 1 (AQP1). V1a and AQP1 gene expression did not correlate; placental V1a mRNA levels were significantly upregulated at 45 and 66+/-1 compared with 27, 100+/-4, and 140 days (term approximately 150 days). V1a mRNA levels were much lower in fetal membranes in which no significant difference across gestation was observed. In situ hybridization histochemistry localized V1a gene expression in the maternal component of the placenta similar to the receptor binding studies using 125I-labeled [d(CH2)5, sarcosine7] vasopressin. No AVP gene expression was observed in the placenta and fetal membranes, which eliminates local AVP production. This increase in V1a expression at 45 and 66+/-1 days of gestation correlates with the period of maximal placental growth in the sheep and suggests that AVP and V1a receptors may play a hitherto unrecognized role in placental growth, differentiation, and/or function, particularly in the deleterious effects of heat stress, early in pregnancy, on fetal growth. PMID- 12724284 TI - Caspase-3 and -6 expression and activation are targeted by hormone action in the rat ventral prostate during the apoptotic cell death process. AB - Although the apoptotic cell death process in the prostate is known to be under the control of androgens, the key components targeted by the hormones remain to be investigated. In the present study, we report that the expression and the activation of the effector caspases-3 and -6 are under the control of testosterone in the adult rat ventral prostate. By using a model of adult castrated rats supplemented (or not) with androgens, we observed an increase in caspase-3 (3-fold) and -6 (4-fold) mRNA (P < 0.0001) and procaspase-3 (32 kDa) and -6 (34 kDa) protein levels by 3 days and 1 wk, respectively, after castration in the ventral prostate. Castration also induced an increase in the activation of the procaspases in the ventral prostate, since active (cleaved) caspase-3 (17 kDa) and -6 (12 kDa) forms reached maximal levels by 1 wk after castration. Testosterone administration to castrated adult rats prevented the increase in caspase-3 and -6 mRNA as well as in procaspase-3 and -6 and active caspase-3 and 6 levels in the ventral prostate lobe. In contrast, no changes were observed in the initiator caspase-8 mRNA and protein (procaspase and active) levels after castration. No changes in caspase-3 and -6 expression and activation were observed in the dorsolateral and anterior prostate lobes after castration and testosterone supplementation. Together, the present results show that testosterone inhibits apoptosis in the ventral prostate by potentially targeting the transcriptional activity of effector caspase-3 and -6 genes (but not of casapase-8 gene) as well as the cleavage of procaspase-3 and -6 into active enzymes. PMID- 12724285 TI - Possible role of cyclooxygenase II in the acquisition of ovarian luteal function in rodents. AB - The development of the corpus luteum (CL), which involves angiogenesis, is essential for the establishment of early pregnancy. We investigated the roles of the prostaglandin synthases cyclooxygenase (COX) I and COX-II in angiogenesis and progesterone production in the newly formed CL, using inhibitors of the COX enzymes and the gonadotropin-induced pseudopregnant rat as a model. Injection of indomethacin, a nonselective COX inhibitor, on the day of ovulation and the following day decreased serum levels of progesterone, as did injection of the selective COX-II inhibitor NS-398. In contrast, a selective COX-I inhibitor, SC 560, had no effect on serum progesterone concentrations. None of the inhibitors had any effect on the weight of the superovulated ovaries or on the synthesis of progesterone by cultured luteal cells. To determine whether changes in angiogenesis are responsible for the decrease in progesterone synthesis, we measured hemoglobin and CD34 levels in luteinized ovaries following injection of COX inhibitors and measured the relative frequency of cells positive for platelet endothelial cell adhesion molecule as a specific marker for endothelial cells. All of these parameters were reduced by the COX-II inhibitors, suggesting that changes in the vasculature are responsible for the decrease in serum progesterone. Histological examination of ovarian corrosion casts indicated that NS-398 inhibited the establishment of luteal capillary vessels following the injection of hCG. The results are consistent with the hypothesis that the activity of COX-II is associated with the formation of functional CL via its stimulation of angiogenesis. PMID- 12724287 TI - Meta, Meta(N) and cyber servers. PMID- 12724286 TI - Human choriocarcinoma cell resistance to natural killer lysis due to defective triggering of natural killer cells. AB - The trophoblast, the outermost layer of the human placenta, lacks expression of the classical human leukocyte antigen (HLA) class I molecules. This prevents allorecognition by T cells but raises the question of what protects the trophoblast from natural killer (NK) cells. In a previous study, we have shown that choriocarcinoma cell (CC) resistance to NK lysis was mainly independent of HLA class I molecules. In the present study, we postulated that CC may prevent activation of NK cells by failing to stimulate their triggering receptors (TR). To test this hypothesis, we evaluated the lysis of JAR and JEG-3 CC after effective cross-linking and activation of NK cells by means of lectins or antibodies. Our results show that NK-resistant CC were sensitive to lysis by unstimulated peripheral blood lymphocytes in the presence of phytohemagglutin (PHA), to antibody-dependent cell cytotoxicity in presence of anti-Tja antibodies, and to monoclonal antibody redirected killing using anti-TR antibodies anti-CD16 and anti-CD244/2B4. Finally, CC fail to express CD48, the ligand for CD244/2B4. These results indicate that the resistance of CC to lysis results primarily from defective NK cell activation, at least partially due to the lack of expression of ligands, such as CD48, involved in the triggering of NK cells. PMID- 12724288 TI - Selection of oligonucleotide probes for protein coding sequences. AB - MOTIVATION: Large arrays of oligonucleotide probes have become popular tools for analyzing RNA expression. However to date most oligo collections contain poorly validated sequences or are biased toward untranslated regions (UTRs). Here we present a strategy for picking oligos for microarrays that focus on a design universe consisting exclusively of protein coding regions. We describe the constraints in oligo design that are imposed by this strategy, as well as a software tool that allows the strategy to be applied broadly. RESULT: In this work we sequentially apply a variety of simple filters to candidate sequences for oligo probes. The primary filter is a rejection of probes that contain contiguous identity with any other sequence in the sample universe that exceeds a pre established threshold length. We find that rejection of oligos that contain 15 bases of perfect match with other sequences in the design universe is a feasible strategy for oligo selection for probe arrays designed to interrogate mammalian RNA populations. Filters to remove sequences with low complexity and predicted poor probe accessibility narrow the candidate probe space only slightly. Rejection based on global sequence alignment is performed as a secondary, rather than primary, test, leading to an algorithm that is computationally efficient. Splice isoforms pose unique challenges and we find that isoform prevalence will for the most part have to be determined by analysis of the patterns of hybridization of partially redundant oligonucleotides. AVAILABILITY: The oligo design program OligoPicker and its source code are freely available at our website. PMID- 12724289 TI - Statistical design of reverse dye microarrays. AB - MOTIVATION: In cDNA microarray experiments all samples are labelled with either Cy3 dye or Cy5 dye. Certain genes exhibit dye bias-a tendency to bind more efficiently to one of the dyes. The common reference design avoids the problem of dye bias by running all arrays 'forward', so that the samples being compared are always labelled with the same dye. But comparison of samples labelled with different dyes is sometimes of interest. In these situations, it is necessary to run some arrays 'reverse'-with the dye labelling reversed-in order to correct for the dye bias. The design of these experiments will impact one's ability to identify genes that are differentially expressed in different tissues or conditions. We address the design issue of how many specimens are needed, how many forward and reverse labelled arrays to perform, and how to optimally assign Cy3 and Cy5 labels to the specimens. RESULTS: We consider three types of experiments for which some reverse labelling is needed: paired samples, samples from two predefined groups, and reference design data when comparison with the reference is of interest. We present simple probability models for the data, derive optimal estimators for relative gene expression, and compare the efficiency of the estimators for a range of designs. In each case, we present the optimal design and sample size formulas. We show that reverse labelling of individual arrays is generally not required. PMID- 12724290 TI - SamCluster: an integrated scheme for automatic discovery of sample classes using gene expression profile. AB - MOTIVATION: Feature (gene) selection can dramatically improve the accuracy of gene expression profile based sample class prediction. Many statistical methods for feature (gene) selection such as stepwise optimization and Monte Carlo simulation have been developed for tissue sample classification. In contrast to class prediction, few statistical and computational methods for feature selection have been applied to clustering algorithms for pattern discovery. RESULTS: An integrated scheme and corresponding program SamCluster for automatic discovery of sample classes based on gene expression profile is presented in this report. The scheme incorporates the feature selection algorithms based on the calculation of CV (coefficient of variation) and t-test into hierarchical clustering and proceeds as follows. At first, the genes with their CV greater than the pre specified threshold are selected for cluster analysis, which results in two putative sample classes. Then, significantly differentially expressed genes in the two putative sample classes with p-values < or = 0.01, 0.05, or 0.1 from t test are selected for further cluster analysis. The above processes were iterated until the two stable sample classes were found. Finally, the consensus sample classes are constructed from the putative classes that are derived from the different CV thresholds, and the best putative sample classes that have the minimum distance between the consensus classes and the putative classes are identified. To evaluate the performance of the feature selection for cluster analysis, the proposed scheme was applied to four expression datasets COLON, LEUKEMIA72, LEUKEMIA38, and OVARIAN. The results show that there are only 5, 1, 0, and 0 samples that have been misclassified, respectively. We conclude that the proposed scheme, SamCluster, is an efficient method for discovery of sample classes using gene expression profile. AVAILABILITY: The related program SamCluster is available upon request or from the web page http://www.sph.uth.tmc.edu:8052/hgc/Downloads.asp. PMID- 12724291 TI - Robust cluster analysis of microarray gene expression data with the number of clusters determined biologically. AB - MOTIVATION: The success of each method of cluster analysis depends on how well its underlying model describes the patterns of expression. Outlier-resistant and distribution-insensitive clustering of genes are robust against violations of model assumptions. RESULTS: A measure of dissimilarity that combines advantages of the Euclidean distance and the correlation coefficient is introduced. The measure can be made robust using a rank order correlation coefficient. A robust graphical method of summarizing the results of cluster analysis and a biological method of determining the number of clusters are also presented. These methods are applied to a public data set, showing that rank-based methods perform better than log-based methods. AVAILABILITY: Software is available from http://www.davidbickel.com. PMID- 12724292 TI - Non-linear normalization and background correction in one-channel cDNA microarray studies. AB - MOTIVATION: Data from one-channel cDNA microarray studies may exhibit poor reproducibility due to spatial heterogeneity, non-linear array-to-array variation and problems in correcting for background. Uncorrected, these phenomena can give rise to misleading conclusions. RESULTS: Spatial heterogeneity may be corrected using two-dimensional loess smoothing (Colantuoni et al., 2002). Non-linear between-array variation may be corrected using an iterative application of one dimensional loess smoothing. A method for background correction using a smoothing function rather than simple subtraction is described. These techniques promote within-array spatial uniformity and between-array reproducibility. Their application is illustrated using data from a study of the effects of an insulin sensitizer, rosiglitazone, on gene expression in white adipose tissue in diabetic db/db mice. They may also be useful with data from two-channel cDNA microarrays and from oligonucleotide arrays. AVAILABILITY: R functions for the methods described are available on request from the author. PMID- 12724294 TI - Reconstructing the temporal ordering of biological samples using microarray data. AB - MOTIVATION: Accurate time series for biological processes are difficult to estimate due to problems of synchronization, temporal sampling and rate heterogeneity. Methods are needed that can utilize multi-dimensional data, such as those resulting from DNA microarray experiments, in order to reconstruct time series from unordered or poorly ordered sets of observations. RESULTS: We present a set of algorithms for estimating temporal orderings from unordered sets of sample elements. The techniques we describe are based on modifications of a minimum-spanning tree calculated from a weighted, undirected graph. We demonstrate the efficacy of our approach by applying these techniques to an artificial data set as well as several gene expression data sets derived from DNA microarray experiments. In addition to estimating orderings, the techniques we describe also provide useful heuristics for assessing relevant properties of sample datasets such as noise and sampling intensity, and we show how a data structure called a PQ-tree can be used to represent uncertainty in a reconstructed ordering. AVAILABILITY: Academic implementations of the ordering algorithms are available as source code (in the programming language Python) on our web site, along with documentation on their use. The artificial 'jelly roll' data set upon which the algorithm was tested is also available from this web site. The publicly available gene expression data may be found at http://genome www.stanford.edu/cellcycle/ and http://caulobacter.stanford.edu/CellCycle/. PMID- 12724293 TI - Gene selection and clustering for time-course and dose-response microarray experiments using order-restricted inference. AB - We propose an algorithm for selecting and clustering genes according to their time-course or dose-response profiles using gene expression data. The proposed algorithm is based on the order-restricted inference methodology developed in statistics. We describe the methodology for time-course experiments although it is applicable to any ordered set of treatments. Candidate temporal profiles are defined in terms of inequalities among mean expression levels at the time points. The proposed algorithm selects genes when they meet a bootstrap-based criterion for statistical significance and assigns each selected gene to the best fitting candidate profile. We illustrate the methodology using data from a cDNA microarray experiment in which a breast cancer cell line was stimulated with estrogen for different time intervals. In this example, our method was able to identify several biologically interesting genes that previous analyses failed to reveal. PMID- 12724295 TI - ALES: cell lineage analysis and mapping of developmental events. AB - MOTIVATION: Animals build their bodies by altering the fates of cells. The way in which they do so is reflected in the topology of cell lineages and the fates of terminal cells. Cell lineages should, therefore, contain information about the molecular events that determined them. Here we introduce new tools for visualizing, manipulating, and extracting the information contained in cell lineages. Our tools enable us to analyze very large cell lineages, where previously analyses have only been carried out on cell lineages no larger than a few dozen cells. RESULTS: Ales (A Lineage Evaluation System) allows the display, evaluation and comparison of cell lineages with the aim of identifying molecular and cellular events underlying development. Ales introduces a series of algorithms that locate putative developmental events. The distribution of these predicted events can then be compared to gene expression patterns or other cellular characteristics. In addition, artificial lineages can be generated, or existing lineages modified, according to a range of models, in order to test hypotheses about lineage evolution. AVAILABILITY: The program can run on any operating system with a compliant Java 2 environment. Ales is free for academic use and can be downloaded from http://mbi.dkfz heidelberg.de/mbi/research/cellsim/ales. PMID- 12724296 TI - MuGeN: simultaneous exploration of multiple genomes and computer analysis results. AB - MOTIVATION: The availability of increasing amounts of sequence data about completely sequenced genomes spurs the development of new methods in the fields of automated annotation, and of comparative genomics. Tools allowing the visualization of results produced by analysis methods, superimposed on possibly annotated sequence data, and enabling synchronized navigation in multiple genomes, provide new means for interactive genome exploration. This kind of visual inspection can be used as a basis to assess the quality of new analysis algorithms, or to discover genome portions to be subjected to in-depth studies. RESULTS: We propose a software package, MuGeN, built for navigating through multiple annotated genomes. It is capable of retrieving annotated sequences in several formats, stored in local files, or available in databases over the network. From these, it then generates an interactive display, or an image file, in most common formats suitable for printing, further editing or integrating in Web pages. Genome maps may be mixed with computer analysis results loaded from XML files, whose format is generic enough to be adapted to a majority of sequence oriented analysis methods. AVAILABILITY: MuGeN is available at http://www mig.jouy.inra.fr/bdsi/MuGeN. PMID- 12724297 TI - A search for H/ACA snoRNAs in yeast using MFE secondary structure prediction. AB - MOTIVATION: Noncoding RNA genes produce functional RNA molecules rather than coding for proteins. One such family is the H/ACA snoRNAs. Unlike the related C/D snoRNAs these have resisted automated detection to date. RESULTS: We develop an algorithm to screen the yeast genome for novel H/ACA snoRNAs. To achieve this, we introduce some new methods for facilitating the search for noncoding RNAs in genomic sequences which are based on properties of predicted minimum free-energy (MFE) secondary structures. The algorithm has been implemented and can be generalized to enable screening of other eukaryote genomes. We find that use of primary sequence alone is insufficient for identifying novel H/ACA snoRNAs. Only the use of secondary structure filters reduces the number of candidates to a manageable size. From genomic context, we identify three strong H/ACA snoRNA candidates. These together with a further 47 candidates obtained by our analysis are being experimentally screened. PMID- 12724298 TI - Improvement of the GenTHREADER method for genomic fold recognition. AB - MOTIVATION: In order to enhance genome annotation, the fully automatic fold recognition method GenTHREADER has been improved and benchmarked. The previous version of GenTHREADER consisted of a simple neural network which was trained to combine sequence alignment score, length information and energy potentials derived from threading into a single score representing the relationship between two proteins, as designated by CATH. The improved version incorporates PSI-BLAST searches, which have been jumpstarted with structural alignment profiles from FSSP, and now also makes use of PSIPRED predicted secondary structure and bi directional scoring in order to calculate the final alignment score. Pairwise potentials and solvation potentials are calculated from the given sequence alignment which are then used as inputs to a multi-layer, feed-forward neural network, along with the alignment score, alignment length and sequence length. The neural network has also been expanded to accommodate the secondary structure element alignment (SSEA) score as an extra input and it is now trained to learn the FSSP Z-score as a measurement of similarity between two proteins. RESULTS: The improvements made to GenTHREADER increase the number of remote homologues that can be detected with a low error rate, implying higher reliability of score, whilst also increasing the quality of the models produced. We find that up to five times as many true positives can be detected with low error rate per query. Total MaxSub score is doubled at low false positive rates using the improved method. AVAILABILITY: http://www.psipred.net. PMID- 12724299 TI - Towards optimal views of proteins. AB - MOTIVATION: Graphical representations of proteins in online databases generally give default views orthogonal to the PDB file coordinate system. These views are often uninformative in terms of protein structure and/or function. Here we discuss the development of a simple automatic algorithm to provide a 'good' view of a protein domain with respect to its structural features. RESULTS: We used dimension reduction with the preservation of topology (using Kohonen's self organising map) to map 3D carbon alpha coordinates into 2D. The original protein structure was then rotated to the view which corresponded most closely to the 2D mapping. This procedure, which we call OVOP, was evaluated in a public blind trial on the web against random views and a 'flattest' view. The OVOP views were consistently rated 'better' than the other views by our volunteers. AVAILABILITY: The source code is available from the OVOP homepage: http://www.sbc.su.se/~oscar/ovop. PMID- 12724300 TI - R/qtl: QTL mapping in experimental crosses. AB - SUMMARY: R/qtl is an extensible, interactive environment for mapping quantitative trait loci (QTLs) in experimental populations derived from inbred lines. It is implemented as an add-on package for the freely-available statistical software, R, and includes functions for estimating genetic maps, identifying genotyping errors, and performing single-QTL and two-dimensional, two-QTL genome scans by multiple methods, with the possible inclusion of covariates. AVAILABILITY: The package is freely available at http://www.biostat.jhsph.edu/~kbroman/qtl. PMID- 12724301 TI - GeneMerge--post-genomic analysis, data mining, and hypothesis testing. AB - SUMMARY: GeneMerge is a web-based and standalone program written in PERL that returns a range of functional and genomic data for a given set of study genes and provides statistical rank scores for over-representation of particular functions or categories in the data set. Functional or categorical data of all kinds can be analyzed with GeneMerge, facilitating regulatory and metabolic pathway analysis, tests of population genetic hypotheses, cross-experiment comparisons, and tests of chromosomal clustering, among others. GeneMerge can perform analyses on a wide variety of genomic data quickly and easily and facilitates both data mining and hypothesis testing. AVAILABILITY: GeneMerge is available free of charge for academic use over the web and for download from: http://www.oeb.harvard.edu/hartl/lab/publications/GeneMerge.html. PMID- 12724302 TI - MARAN: normalizing micro-array data. AB - SUMMARY: MARAN is a web-based application for normalizing microarray data. MARAN comprises a generic ANOVA model, an option for Loess fitting prior to ANOVA analysis, and a module for selecting genes with significantly changing expression. AVAILABILITY: http://www.esat.kuleuven.ac.be/maran/. PMID- 12724303 TI - Do current sequence analysis algorithms disclose multifunctional (moonlighting) proteins? AB - SUMMARY: A number of multifunctional, 'moonlighting', proteins have been analyzed by different current programs to test whether they identify both functions. PSI BLAST and PRODOM perform best in predicting the alternative function. PMID- 12724305 TI - SCide: identification of stabilization centers in proteins. AB - SUMMARY: SCide is a program to identify stabilization centers from known protein structures. These are residues involved in cooperative long-range contacts, which can be formed between various regions of a single polypeptide chain, or they can belong to different peptides or polypeptides in a complex. The server takes a PDB file as an input, and the result is presented in graphical or text format. AVAILABILITY: SCide is available on the web at http://www.enzim.hu/scide. The source code can be obtained from the authors on request. PMID- 12724304 TI - BioEditor-simplifying macromolecular structure annotation. AB - SUMMARY: BioEditor is an application to enable scientists and educators to prepare and present structure annotations containing formatted text, graphics, sequence data, and interactive molecular views. It is intended to bridge the gap between printed journal articles and Internet presentation formats. BioEditor is relevant in the era of structural genomics, where annotation and publication could become the rate determining step in structure determination. AVAILABILITY: BioEditor is available at http://bioeditor.sdsc.edu. The Web site includes the latest version of the software for Microsoft Windows, including documentation, the opportunity to submit bug reports and suggestions, example documentaries prepared with BioEditor and a repository where users can submit documentaries for posting to the site. PMID- 12724306 TI - BioQuery: an object framework for building queries to biomedical databases. AB - SUMMARY: BioQuery is an application that helps scientists automate database searches. Users can build and store queries to public biomedical databases, and receive periodic updates on the results of those queries when new data is available. The application is implemented on a portable object framework that can provide database-searching capability to other applications. This framework is easily extensible, allowing users to develop plug-ins that provide access to new databases. BioQuery thus provides end-users with a complete database searching interface and updating service, and gives developers a toolkit to provide database-searching capability to their applications. AVAILABILITY: Free to all users: http://www.bioquery.org. PMID- 12724307 TI - IPPRED: server for proteins interactions inference. AB - SUMMARY: IPPRED is a web based server to infer protein-protein interactions through homology search between candidate proteins and those described as interacting. This simple inference allows to propose or to validate potential interactions. AVAILABILITY: IPPRED is freely available at http://cbi.labri.fr/outils/ippred/. PMID- 12724308 TI - Protein kinase C beta is required for human monocyte chemotaxis to MCP-1. AB - Monocyte chemoattractant protein 1 (MCP-1) is important in attracting monocytes to sites of inflammation. Using predominantly pharmacological approaches, prior studies have indicated that serine/threonine kinases are involved in the MCP-1 induced signaling pathways. We report here that there is substantial inhibition of MCP-1-stimulated chemotaxis of human monocytes treated with inhibitors selective for the subset of serine/threonine kinases, protein kinase C (PKC). Selective inhibitors of PKC such as GF109203X and Calphostin C both caused approximately 80% inhibition of chemotaxis. Because these pharmacological inhibitors do not specifically inhibit individual PKC isoforms, we chose to use antisense oligodeoxyribonucleotides (ODN) to specifically reduce PKC isoform expression, first by inhibiting expression of the conventional PKC family, and next by using specific antisense ODN for PKCalpha and PKCbeta. Conventional PKC antisense ODN treatment completely and significantly inhibited monocyte chemotaxis to MCP-1, whereas sense-control ODN caused no significant inhibition. PKCbeta-antisense ODN caused 89.2% inhibition of chemotaxis at its highest dose. In contrast, PKCbeta-sense ODN and PKCalpha-antisense and -sense ODN were without effect. Further studies evaluating the calcium response that is triggered upon MCP-1 interaction with its receptor, CCR2, indicate that this response is not altered by antisense or sense ODN treatment, thus supporting our hypothesis that PKCbeta is critical for post-receptor signal transduction downstream of the immediate calcium signal. These data contribute to our developing understanding of the signal transduction pathways involved in the chemotactic response of human monocytes to MCP-1 and uniquely identify the requirement for the PKCbeta isoform in this important process. PMID- 12724310 TI - Site-directed mutagenesis of benzalacetone synthase. The role of the Phe215 in plant type III polyketide synthases. AB - Benzalacetone synthase (BAS) and chalcone synthase (CHS) are plant-specific type III polyketide synthases (PKSs) that share approximately 70% amino acid sequence identity. BAS catalyzes a one-step decarboxylative condensation of 4-coumaroyl CoA with malonyl-CoA to produce a diketide benzalacetone, whereas CHS performs sequential condensations with three malonyl-CoA to generate a tetraketide chalcone. A homology model suggested that BAS has the same overall fold as CHS with cavity volume almost as large as that of CHS. One of the most characteristic features is that Rheum palmatum BAS lacks active site Phe-215; the residues 214LF conserved in type III PKSs are uniquely replaced by IL. Our observation that the BAS I214L/L215F mutant exhibited chalcone-forming activity in a pH-dependent manner supported a hypothesis that the absence of Phe-215 in BAS accounts for the interruption of the polyketide chain elongation at the diketide stage. On the other hand, Phe-215 mutants of Scutellaria baicalensis CHS (L214I/F215L, F215W, F215Y, F215S, F215A, F215H, and F215C) afforded increased levels of truncated products; however, none of them generated benzalacetone. These results confirmed the critical role of Phe-215 in the polyketide formation reactions and provided structural basis for understanding the structure-function relationship of the plant type III PKSs. PMID- 12724309 TI - Tyrosinase maturation and oligomerization in the endoplasmic reticulum require a melanocyte-specific factor. AB - Tyrosinase is a glycoprotein responsible for the synthesis of melanin in melanocytes. A large number of mutations have been identified in tyrosinase, with many leading to its misfolding, endoplasmic reticulum (ER) retention, and degradation. Here we describe the folding and maturation of human tyrosinase (TYR) using an in vitro translation system coupled with ER-derived microsomes or with semipermeabilized cells, as an intact ER source. TYR remained misfolded as determined by its sensitivity to trypsin digestion and its persistent interaction with the ER resident lectin chaperones calnexin and calreticulin when produced in ER-derived microsomes or nonmelanocytic semipermeabilized cells. However, when TYR was translocated into semipermeabilized melanocytes, chaperone interactions were transient, maturation progressed to a trypsin-resistant state, and a TYR homodimer was formed. The use of semipermeabilized mouse melanocytes defective for tyrosinase or other melanocyte-specific proteins as the ER source indicated that proper TYR maturation and oligomerization were greatly aided by the presence of wild type tyrosinase and tyrosinase-related protein 1. These findings suggested that oligomerization is a step in proper TYR maturation within the ER that requires melanocyte-specific factors. PMID- 12724311 TI - Ca2+-dependent potentiation of the nonselective cation channel TRPV4 is mediated by a C-terminal calmodulin binding site. AB - Most Ca2+-permeable ion channels are inhibited by increases in the intracellular Ca2+ concentration ([Ca2+]i), thus preventing potentially deleterious rises in [Ca2+]i. In this study, we demonstrate that currents through the osmo-, heat- and phorbol ester-sensitive, Ca2+-permeable nonselective cation channel TRPV4 are potentiated by intracellular Ca2+. Spontaneous TRPV4 currents and currents stimulated by hypotonic solutions or phorbol esters were reduced strongly at all potentials in the absence of extracellular Ca2+. The other permeant divalent cations Ba2+ and Sr2+ were less effective than Ca2+ in supporting channel activity. An intracellular site of Ca2+ action was supported by the parallel decrease in spontaneous currents and [Ca2+]i on removal of extracellular Ca2+ and the ability of Ca2+ release from intracellular stores to restore TRPV4 activity in the absence of extracellular Ca2+. During TRPV4 activation by hypotonic solutions or phorbol esters, Ca2+ entry through the channel increased the rate and extent of channel activation. Currents were also potentiated by ionomycin in the presence of extracellular Ca2+. Ca2+-dependent potentiation of TRPV4 was often followed by inhibition. By mutagenesis, we localized the structural determinant of Ca2+-dependent potentiation to an intracellular, C-terminal calmodulin binding domain. This domain binds calmodulin in a Ca2+-dependent manner. TRPV4 mutants that did not bind calmodulin lacked Ca2+-dependent potentiation. We conclude that TRPV4 activity is tightly controlled by intracellular Ca2+. Ca2+ entry increases both the rate and extent of channel activation by a calmodulin-dependent mechanism. Excessive increases in [Ca2+]i via TRPV4 are prevented by a Ca2+-dependent negative feedback mechanism. PMID- 12724312 TI - Ganglioside GM3 inhibits matrix metalloproteinase-9 activation and disrupts its association with integrin. AB - Gangliosides GM3 and GT1b both inhibit epithelial cell adhesion and migration on fibronectin. GT1b binds to integrin alpha5beta1 and blocks the integrin fibronectin interaction; GM3 does not interact with integrin, and its effect is poorly understood. We evaluated the effects of endogenous modulation of GM3 expression on epithelial cell motility on several matrices and the mechanism of these effects. Endogenous accumulation of GM3 decreased cell migration on fibronectin, types I, IV, and VII collagen matrices; depletion of GM3 dramatically increased cell migration, regardless of matrix. GM3 overexpression and depletion in vitro correlated inversely with the expression and activity of matrix metalloproteinase-9; consistently, the cell migration stimulated by GM3 depletion is reversed by inhibition of matrix metalloproteinase-9 activity. Accumulation and depletion of GM3 in epithelial cells grown on fibronectin also correlated inversely with epidermal growth factor receptor and mitogen activated protein kinase phosphorylation and with Jun expression. Ganglioside depletion facilitated the co-immunoprecipitation of matrix metal-loproteinase-9 and integrin alpha5beta1, while endogenous accumulation of GM3, but not GT1b, blocked the co-immunoprecipitation. These data suggest modulation of epidermal growth factor receptor signaling and dissociation of integrin/matrix metalloproteinase-9 as mechanisms for the GM3-induced effects on matrix metalloproteinase-9 function. PMID- 12724313 TI - Roles of the tetratricopeptide repeat domain in O-GlcNAc transferase targeting and protein substrate specificity. AB - The abundant and dynamic post-translational modification of nuclear and cytosolic proteins by beta-O-linked N-acetylglucosamine (O-GlcNAc) is catalyzed by O-GlcNAc transferase (OGT). Recently, we reported the identification of a novel family of OGT-interacting proteins (OIPs) that interact strongly with the tetratricopeptide repeat (TPR) domain of OGT (Iyer, S. P., Akimoto, Y., and Hart, G. W. (2003) J. Biol. Chem. 278, 5399-5409). Members of this family are modified by O-GlcNAc and are excellent substrates of OGT. Here, we further investigated the role of the TPR domain in the O-GlcNAcylation of OIP106, one of the members of this OIP family. Using N-terminal deletions, we first identified the region of OIP106 that binds OGT, termed the OGT-interacting domain (OID). Deletion analysis indicated that TPRs 2-6 of OGT interact with the OID of OIP106. The apparent Km of OGT for the OID of OIP106 is 3.35 microm. Unlike small peptide substrates, glycosylation of the OID was dependent upon its interaction with the first 6 TPRs of OGT. Furthermore, the isolated TPR domain of OGT competitively inhibited glycosylation of the OID protein, but did not inhibit glycosylation of a 12-amino acid casein kinase II peptide substrate, providing kinetic evidence for the role of the TPR domain as a protein substrate docking site. Additionally, both the OID of OIP106 and nucleoporin p62 competed with each other for glycosylation by OGT. These studies support the model that the catalytic subunit of OGT achieves both high specificity and a remarkable diversity of substrates by complexing with a variety of targeting proteins via its TPR protein-protein interaction domains. PMID- 12724314 TI - Identification and characterization of a novel p300-mediated p53 acetylation site, lysine 305. AB - Post-translational modifications serve as important regulatory elements in modulating the transcriptional activity of the tumor suppressor protein p53. We have previously reported a tandem mass spectrometry-based method (viz. selected ion tracing analysis) that can be applied to the identification of phosphopeptides as well as exact mapping of the phosphorylated residues within. In this study, we describe the application of the same strategy for the identification of p300 acetyltransferase-mediated acetylation sites on p53. Consistent with the previous finding, lysines 370, 372, 373, 381, and 382 were detected by this modified selected ion tracing method as the target sites of p300 in vitro. Moreover, two novel acetylation sites, Lys-292 and Lys-305, were also found. Immunoblotting using anti-acetyl-Lys-305 antibody confirmed this discovery and demonstrated that Lys-305 could be acetylated by p300 both in vitro and in vivo. We also show that an alanine or glutamine substitution at Lys-305 (K305A or K305Q) suppressed the transcriptional activity of p53, whereas an arginine mutation (K305R) increased the transcriptional activity. Thus, p300 may further regulate the transcriptional activity of p53 through a novel acetylation site, Lys-305. PMID- 12724315 TI - Direct demonstration of involvement of protein kinase Calpha in the Ca2+-induced platelet aggregation. AB - Platelets play critical roles in hemostasis and thrombosis through their aggregation following activation of integrin alphaIIbbeta3. However, the molecular mechanism of the integrin activation inside platelets remains largely unknown. Pharmacological experiments have demonstrated that protein kinase C (PKC) plays an important role in platelet aggregation. Because PKC inhibitors can have multiple substrates and given that non-PKC-phorbol ester-binding signaling molecules have been demonstrated to play important roles, the precise involvement of PKC in cellular functions requires re-evaluation. Here, we have established an assay for analyzing the Ca2+-induced aggregation of permeabilized platelets. The aggregation of platelets was inhibited by the addition of the arginine-glycine aspartate-serine peptide, an integrin-binding peptide inhibitor of alphaIIbbeta3, suggesting that the aggregation was mediated by the integrin. The aggregation was also dependent on exogenous ATP and platelet cytosol, indicating the existence of essential cytosolic factors required for the aggregation. To examine the role of PKC in the aggregation assay, we immunodepleted PKCalpha and beta from the cytosol. The PKC-depleted cytosol lost the aggregation-supporting activity, which was recovered by the addition of purified PKCalpha. Furthermore, the addition of purified PKCalpha in the absence of cytosol did not support the aggregation, whereas the cytosol containing less PKC supported it efficiently, suggesting that additional factors besides PKC would also be required. Thus, we directly demonstrated that PKCalpha is involved in the regulation of Ca2+-induced platelet aggregation. PMID- 12724316 TI - Desensitization of signaling by oncostatin M in human vascular cells involves cytoplasmic Tyr residue 759 in gp130 but is not mediated by either Src homology 2 domain-containing tyrosine phosphatase 2 or suppressor of cytokine signaling 3. AB - Oncostatin M (OnM) signals through cell surface receptors, which utilize the gp130 subunit. In cultured human umbilical vein endothelial cells (HUVEC), OnM transiently elevates mRNA encoding for suppressor of cytokine signaling-3 (SOCS 3). By 1 h of OnM treatment, HUVEC become refractory to the restimulation by OnM, measured as failure to reinduce SOCS-3 mRNA. OnM-induced desensitization also prevents responses to other gp130-signaling cytokines (e.g. leukemia inhibitory factor and interleukin 11). OnM treatment does not affect gp130 expression levels and desensitizes signaling mediated by a transduced chimeric receptor containing extracellular domains of platelet-derived growth factor receptor-beta (PDGFRbeta) and the cytoplasmic region of gp130. Interestingly, a chimeric PDGFRbeta-gp130 mutant receptor, in which intracellular Tyr residue 759 of gp130 is replaced by a Phe residue, mediates prolonged signaling and is not cross-desensitized by OnM. Phospho-Tyr759 is the binding site for both SOCS-3 and for Src homology domain 2 containing tyrosine phosphatase 2 (SHP-2). In human aortic smooth muscle cells, neither prevention of SOCS-3 protein induction, using STAT3 or SOCS-3 antisense, nor prevention of SHP-2 expression, also with antisense, ablates desensitization. These data suggest that desensitization of vascular cells to OnM is mediated in trans and involves Tyr residue 759 in gp130 but is not mediated by either SOCS-3 or SHP-2, the only two proteins currently known to bind to gp130 at this site. PMID- 12724317 TI - StAR-related lipid transfer (START) proteins: mediators of intracellular lipid metabolism. PMID- 12724318 TI - Enrichment for histone H3 lysine 9 methylation at Alu repeats in human cells. AB - The aim of this study was to identify in human cells common targets of histone H3 lysine 9 (H3-Lys9) methylation, a modification that is generally associated with gene silencing. After chromatin immunoprecipitation using an H3-Lys9 methylated antibody, we cloned the recovered DNA and sequenced 47 independent clones. Of these, 38 clones (81%) contained repetitive elements, either short interspersed transposable element (SINE or Alu elements), long terminal repeat (LTR), long interspersed transposable element (LINE), or satellite region (ALR/Alpha) DNA, and three additional clones were near Alu elements. Further characterization of these repetitive elements revealed that 32 clones (68%) were Alu repeats, corresponding to both old Alu (23 clones) and young Alu (9 clones) subfamilies. Association of H3-Lys9 methylation was confirmed by chromatin immunoprecipitation PCR using conserved Alu primers. In addition, we randomly selected 5 Alu repeats from the recovered clones and confirmed association with H3-Lys9 by PCR using primer sets flanking the Alu elements. Treatment with the DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine rapidly decreased the level of H3-Lys9 methylation in the Alu elements, suggesting that H3-Lys9 methylation may be related to the suppression of Alu elements through DNA methylation. Thus H3-Lys9 methylation is enriched at human repetitive elements, particularly Alu elements, and may play a role in the suppression of recombination by these elements. PMID- 12724319 TI - The spatial organization of apolipoprotein A-I on the edge of discoidal high density lipoprotein particles: a mass specrometry study. AB - The three-dimensional structure of human apoA-I on nascent, discoidal HDL particles has been debated extensively over the past 25 years. Recent evidence has demonstrated that the alpha-helical domains of apoA-I are arranged in a belt like orientation with the long axis of the helices perpendicular to the phospholipid acyl chains on the disc edge. However, experimental information on the spatial relationships between apoA-I molecules on the disc is lacking. To address this issue, we have taken advantage of recent advances in mass spectrometry technology combined with cleavable cross-linking chemistry to derive a set of distance constraints suitable for testing apoA-I structural models. We generated highly homogeneous, reconstituted HDL particles containing two molecules of apoA-I. These were treated with a thiol-cleavable cross-linking agent, which covalently joined Lys residues in close proximity within or between molecules of apoA-I in the disc. The cross-linked discs were then exhaustively trypsinized to generate a discrete population of peptides. The resulting peptides were analyzed by liquid chromatography/mass spectrometry before and after cleavage of the cross-links, and resulting peaks were identified based on the theoretical tryptic cleavage of apoA-I. We identified at least 8 intramolecular and 7 intermolecular cross-links in the particle. The distance constraints are used to analyze three current models of apoA-I structure. The results strongly support the presence of the salt-bridge interactions that were predicted to occur in the "double belt" model of apoA-I, but a helical hairpin model containing the same salt-bridge docking interface is also consistent with the data. PMID- 12724320 TI - Identification of p2y9/GPR23 as a novel G protein-coupled receptor for lysophosphatidic acid, structurally distant from the Edg family. AB - Lysophosphatidic acid (LPA) is a bioactive lipid mediator with diverse physiological and pathological actions on many types of cells. LPA has been widely considered to elicit its biological functions through three types of G protein-coupled receptors, Edg-2 (endothelial cell differentiation gene 2)/LPA1/vzg-1 (ventricular zone gene-1), Edg-4/LPA2, and Edg-7/LPA3. We identified an orphan G protein-coupled receptor, p2y9/GPR23, as the fourth LPA receptor (LPA4). Membrane fractions of RH7777 cells transiently expressing p2y9/GPR23 displayed a specific binding for 1-oleoyl-LPA with a Kd value of around 45 nm. Competition binding and reporter gene assays showed that p2y9/GPR23 preferred structural analogs of LPA with a rank order of 1-oleoyl- > 1-stearoyl- > 1-palmitoyl- > 1-myristoyl- > 1-alkyl- > 1-alkenyl-LPA. In Chinese hamster ovary cells expressing p2y9/GPR23, 1-oleoyl-LPA induced an increase in intracellular Ca2+ concentration and stimulated adenylyl cyclase activity. Quantitative real-time PCR demonstrated that mRNA of p2y9/GPR23 was significantly abundant in ovary compared with other tissues. Interestingly, p2y9/GPR23 shares only 20-24% amino acid identities with Edg-2/LPA1, Edg-4/LPA2, and Edg-7/LPA3, and phylogenetic analysis also shows that p2y9/GPR23 is far distant from the Edg family. These facts suggest that p2y9/GPR23 has evolved from different ancestor sequences from the Edg family. PMID- 12724321 TI - Direct interaction of subunits a and b of the F0 complex of Escherichia coli ATP synthase by forming an ab2 subcomplex. AB - The addition of a His6 tag to the N terminus of subunit a of the F0 complex of the Escherichia coli ATP synthase allowed the purification of an ab2 subcomplex after solubilization of membranes with n-dodecyl-beta-d-maltoside and subsequent nickel-nitrilotriacetic acid affinity chromatography. After co-reconstitution of the ab2 subcomplex with purified subunit c, passive proton translocation rates as well as coupled ATPase activities after binding of F1 were measured that were comparable with those of wild type F0. The interaction between subunits a and b, which has been shown to be stoichiometric and functional, is not triggered by any cross-linking reagent and therefore reflects subunit interactions occurring within the F0 complex in vivo. PMID- 12724322 TI - Bruton's tyrosine kinase is a Toll/interleukin-1 receptor domain-binding protein that participates in nuclear factor kappaB activation by Toll-like receptor 4. AB - In this study we have identified members of the Toll-like receptor (TLR) family (namely, TLRs 4, 6, 8, and 9) as proteins to which the intracellular protein tyrosine kinase, Bruton's tyrosine kinase (Btk), binds. Detailed analysis of the interaction between Btk and TLR8 demonstrates that the presence of both Box 2 and 3 motifs in the Toll/interleukin-1 receptor domain was required for the interaction. Furthermore, co-immunoprecipitation experiments revealed that Btk can also interact with key proteins involved in TLR4 signal transduction, namely, MyD88, Mal (MyD88 adapter-like protein), and interleukin-1 receptor-associated kinase-1, but not TRAF-6. The ability of Btk to interact with TLR4 and Mal suggests a role for Btk in lipopolysaccharide (LPS) signal transduction. Stimulation of the human monocytic cell line THP-1 with LPS resulted in an increase in the level of tyrosine phosphorylation of Btk (indicative of activation). The autokinase activity of Btk was also stimulated after LPS stimulation. In addition, a dominant negative form of Btk inhibited TLR4-mediated activation of a nuclear factor kappaB (NFkappaB)-dependent reporter gene in HEK293 cells as well as LPS-induced activation of NFkappaB in the astrocytoma cell line U373 and the monocytic cell line RAW264.7. Further investigation revealed that the Btk-specific inhibitor, LFM-A13, inhibited the activation of NFkappaB by LPS in THP-1 cells. Our findings implicate Btk as a Toll/interleukin 1 receptor domain-binding protein that is important for NFkappaB activation by TLR4. PMID- 12724323 TI - Comparison of prostaglandin F2alpha, bimatoprost (prostamide), and butaprost (EP2 agonist) on Cyr61 and connective tissue growth factor gene expression. AB - Connective tissue growth factor (CTGF) and Cyr61 (cysteine-rich angiogenic protein 61) are members of the CCN gene family that encode multifunctional, extracellular matrix-associated signaling proteins. Because the mechanism of action of certain anti-glaucoma drugs involves extracellular matrix remodeling of ocular ciliary muscle, with a resultant increase in drainage of aqueous humor from the eye, we compared the effects of three pharmacologically distinct ocular hypotensive agents on Cyr61 and CTGF gene expression. Thus, prostaglandin F2alpha (PGF2alpha) (FP receptor agonist), Butaprost (EP2 receptor agonist), and Bimatoprost (a prostamide) were compared. Using Affymetrix gene chip technology, we first identified that PGF2alpha dramatically up-regulated Cyr61 and CTGF mRNA expression in HEK 293/EBNA cells (hFP-HEK 293/EBNA). Northern blot further confirmed the Cyr61 and CTGF up-regulation is in a dose- and time-dependent manner. PGF2alpha-induced up-regulation of Cyr61 appeared to exclusively involve the Rho pathway, and up-regulation of CTGF was via multiple intracellular pathways. Because prostamide receptors are, to date, defined only at the pharmacological level, Bimatoprost effects on Cyr61 and CTGF were studied in the isolated feline iris sphincter preparation, a tissue highly responsive to prostamides. Both PGF2alpha and Bimatoprost up-regulated Cyr61 mRNA expression in the cat iris tissue. Only PGF2alpha up-regulated CTGF mRNA expression in the cat iris. Therefore, PGF2alpha and Bimatoprost appear to interact with different receptors populations in the cat iris, according to their markedly different effects on CTGF. Activation of prostaglandin EP2 receptors (Gs-coupled) also up regulated Cyr61 but not CTGF mRNA expression in the isolated cat iris. Similar data were observed in human primary ciliary smooth muscle cells. Thus, despite quite different signal transduction pathways, FP receptor stimulation up regulates CTGF and Cyr61. The prostamide analog Bimatoprost and an EP2-selective agonist affects only Cyr61. PMID- 12724324 TI - Execution of macrophage apoptosis by Mycobacterium avium through apoptosis signal regulating kinase 1/p38 mitogen-activated protein kinase signaling and caspase 8 activation. AB - Macrophage apoptosis is an important component of the innate immune defense machinery (against pathogenic mycobacteria) responsible for limiting bacillary viability. However, little is known about the mechanism of how apoptosis is executed in mycobacteria-infected macrophages. Apoptosis signal-regulating kinase 1 (ASK1) was activated in Mycobacterium avium-treated macrophages and in turn activated p38 mitogen-activated protein (MAP) kinase. M. avium-induced macrophage cell death could be blocked in cells transfected with a catalytically inactive mutant of ASK1 or with dominant negative p38 MAP kinase arguing in favor of a central role of ASK1/p38 MAP kinase signaling in apoptosis of macrophages challenged with M. avium. ASK1/p38 MAP kinase signaling was linked to the activation of caspase 8. At the same time, M. avium triggered caspase 8 activation, and cell death occurred in a Fas-associated death domain (FADD) dependent manner. The death signal induced upon caspase 8 activation linked to mitochondrial death signaling through the formation of truncated Bid (t-Bid), its translocation to the mitochondria and release of cytochrome c. Caspase 8 inhibitor (z-IETD-FMK) could block the release of cytochrome c as well as the activation of caspases 9 and 3. The final steps of apoptosis probably involved caspases 9 and 3, since inhibitors of both caspases could block cell death. Of foremost interest in the present study was the finding that ASK1/p38 signaling was essential for caspase 8 activation linked to M. avium-induced death signaling. This work provides the first elucidation of a signaling pathway in which ASK1 plays a central role in innate immunity. PMID- 12724326 TI - Dynamic traffic through the recycling compartment couples the metal transporter Nramp2 (DMT1) with the transferrin receptor. AB - Nramp2 (natural resistance-associated macrophage protein 2, also called DMT1 and Slc11a2) is a proton-dependent cation transporter, which plays a central role in iron homeostasis. To study the subcellular distribution and dynamics of the transporter, we generated a construct encoding the long splice variant of Nramp2 (isoform II) tagged with the hemagglutinin epitope on a predicted extracellular loop. Cells stably transfected with this construct revealed the presence of Nramp2 in both the plasma membrane and in an endomembrane compartment. By labeling the exofacial epitope with a pH-sensitive fluorescent indicator, we were able to establish that this variant of Nramp2 resides in a vesicular compartment with an acidic lumen (pH 6.2) and that acidification was maintained by vacuolar type ATPases. Dual labeling experiments identified this compartment as sorting and recycling endosomes. Kinetic studies by surface labeling with 125I-labeled antibodies established that the fraction of endomembrane Nramp2 was approximately equal to that on the cell surface. The two components are in dynamic equilibrium: surface transporters are internalized continuously via a clathrin and dynamin dependent process, whereas endosomal Nramp2 is recycled to the plasmalemma by a phosphatidylinositol 3-kinase-dependent exocytic process. Depletion of cholesterol had no discernible effect on Nramp2 internalization, suggesting that rafts or caveolae are not essential. Because the pH at the cell surface and in endosomes differs by >or=1 unit, the rates of transport of Nramp2 at the surface and in endomembrane compartments will differ drastically. Their subcellular colocalization and parallel trafficking suggest that Nramp2 and transferrin receptors are functionally coupled to effect pH-dependent iron uptake across the endosomal membrane. PMID- 12724325 TI - Synergistic cooperation between the beta-catenin signaling pathway and steroidogenic factor 1 in the activation of the Mullerian inhibiting substance type II receptor. AB - Mullerian inhibiting substance type II receptor (MISRII) is a member of the transforming growth factor-beta superfamily. Mutations in mullerian inhibiting substance (MIS) or MISRII cause male sexual abnormalities, persistent mullerian duct syndrome, and pseudohermaphroditism. The spatial and temporal regulation of MIS and MISRII is important for its biological action. Male Wnt7a mutant mice do not undergo regression of mullerian ducts. Here we showed that the canonical Wnt signaling pathway regulated MISRII. The promoter MISRII was activated by beta catenin expression, and this activation was dependent on TCF4-binding sites. The nuclear receptor superfamily member steroidogenic factor 1 (SF1) synergistically activated the MISRII promoter with beta-catenin. APC, a negative regulator of Wnt signaling, decreased SF1-mediated activation of the MISRII promoter in the colon carcinoma cell line SW480. We also showed a direct physical interaction between beta-catenin and SF1 by co-immunoprecipitation. Thus, our findings suggest that MISRII is a developmental target of Wnt7a signaling for mullerian duct regression during sexual differentiation. PMID- 12724327 TI - GIPC interacts with the beta1-adrenergic receptor and regulates beta1-adrenergic receptor-mediated ERK activation. AB - Beta1-adrenergic receptors, expressed at high levels in the human heart, have a carboxyl-terminal ESKV motif that can directly interact with PDZ domain containing proteins. Using the beta1-adrenergic receptor carboxyl terminus as bait, we identified the novel beta1-adrenergic receptor-binding partner GIPC in a yeast two-hybrid screen of a human heart cDNA library. Here we demonstrate that the PDZ domain-containing protein, GIPC, co-immunoprecipitates with the beta1 adrenergic receptor in COS-7 cells. Essential for this interaction is the Ser residue of the beta1-adrenergic receptor carboxyl-terminal ESKV motif. Our data also demonstrate that beta1-adrenergic receptor stimulation activates the mitogen activated protein kinase, ERK1/2. beta1-adrenergic receptor-mediated ERK1/2 activation was inhibited by pertussis toxin, implicating Gi, and was substantially decreased by the expression of GIPC. Expression of GIPC had no observable effect on beta1-adrenergic receptor sequestration or receptor-mediated cAMP accumulation. This GIPC effect was specific for the beta1-adrenergic receptor and was dependent on an intact PDZ binding motif. These data suggest that GIPC can regulate beta1-adrenergic receptor-stimulated, Gi-mediated, ERK activation while having no effect on receptor internalization or Gs-mediated cAMP signaling. PMID- 12724328 TI - Calcium dependence of the interaction between calmodulin and anthrax edema factor. AB - Edema factor (EF), a toxin from Bacillus anthracis (anthrax), possesses adenylyl cyclase activity and requires the ubiquitous Ca2+-sensor calmodulin (CaM) for activity. CaM can exist in three major structural states: an apo state with no Ca2+ bound, a two Ca2+ state with its C-terminal domain Ca2+-loaded, and a four Ca2+ state in which the lower Ca2+ affinity N-terminal domain is also ligated. Here, the interaction of EF with the three Ca2+ states of CaM has been examined by NMR spectroscopy and changes in the Ca2+ affinity of CaM in the presence of EF have been determined by flow dialysis. Backbone chemical shift perturbations of CaM show that EF interacts weakly with the N-terminal domain of apoCaM. The C terminal CaM domain only engages in the interaction upon Ca2+ ligation, rendering the overall interaction much tighter. In the presence of EF, the C-terminal domain binds Ca2+ with higher affinity, but loses binding cooperativity, whereas the N-terminal domain exhibits strongly reduced Ca2+ affinity. As judged by chemical shift differences, the N-terminal CaM domain remains bound to EF upon subsequent Ca2+ ligation. This Ca2+ dependence of the EF-CaM interaction differs from that observed for most other CaM targets, which normally interact only with the Ca2+-bound CaM domains and become active following the transition to the four Ca2+ state. PMID- 12724329 TI - Interaction of glycoprotein H of human herpesvirus 6 with the cellular receptor CD46. AB - Human herpesvirus 6 (HHV-6) employs the complement regulator CD46 (membrane cofactor protein) as a receptor for fusion and entry into target cells. Like other known herpesviruses, HHV-6 encodes multiple glycoproteins, several of which have been implicated in the entry process. In this report, we present evidence that glycoprotein H (gH) is the viral component responsible for binding to CD46. Antibodies to CD46 co-immunoprecipitated an approximately 110-kDa protein band specifically associated with HHV-6-infected cells. This protein was identified as gH by selective depletion with an anti-gH monoclonal antibody, as well as by immunoblot analysis with a rabbit hyperimmune serum directed against a gH synthetic peptide. In reciprocal experiments, a monoclonal antibody against HHV-6 gH was found to co-immunoprecipitate CD46. Studies using monoclonal antibodies directed against specific CD46 domains, as well as engineered constructs lacking defined CD46 regions, demonstrated a close correspondence between the CD46 domains involved in the interaction with gH and those previously shown to be critical for HHV-6 fusion (i.e. short consensus repeats 2 and 3). PMID- 12724330 TI - The inhibitor thiomandelic acid binds to both metal ions in metallo-beta lactamase and induces positive cooperativity in metal binding. AB - Thiomandelic acid is a simple, broad spectrum, and reasonably potent inhibitor of metallo-beta-lactamases, enzymes that mediate resistance to beta-lactam antibiotics. We report studies by NMR and perturbed angular correlation (PAC) spectroscopy of the mode of binding of the R and S enantiomers of thiomandelic acid, focusing on their interaction with the two metal ions in cadmium substituted Bacillus cereus metallo-beta-lactamase. The 113Cd resonances are specifically assigned to the metals in the two individual sites on the protein by using 113Cd-edited 1H NMR spectra. Each enantiomer of thiomandelate produces large downfield shifts of both 113Cd resonances and changes in the PAC spectra, which indicate that they bind such that the thiol of the inhibitor bridges between the two metals. For R-thiomandelate, this is unambiguously confirmed by the observation of scalar coupling between Halpha of the inhibitor and both cadmium ions. The NMR and PAC spectra reveal that the two chiral forms of the inhibitor differ in the details of their coordination geometry. The complex with R-thiomandelate, but not that with the S-enantiomer, shows evidence in the PAC spectra of a dynamic process in the nanosecond time regime, the possible nature of which is discussed. The thiomandelate complex of the mononuclear enzyme can be detected only at low metal to enzyme stoichiometry; the relative populations of mononuclear and binuclear enzyme as a function of cadmium concentration provide clear evidence for positive cooperativity in metal ion binding in the presence of the inhibitor, in contrast to the negative cooperativity observed in the free enzyme. PMID- 12724331 TI - Three distinct epitopes on the extracellular face of the glucagon receptor determine specificity for the glucagon amino terminus. AB - The glucagon and glucagon-like peptide-1 (GLP-1) receptors are homologous family B seven-transmembrane (7TM) G protein-coupled receptors, and they selectively recognize the homologous peptide hormones glucagon (29 amino acids) and GLP-1 (30 31 amino acids), respectively. The amino-terminal extracellular domain of the glucagon and GLP-1 receptors (140-150 amino acids) determines specificity for the carboxyl terminus of glucagon and GLP-1, respectively. In addition, the glucagon receptor core domain (7TM helices and connecting loops) strongly determines specificity for the glucagon amino terminus. Only 4 of 15 residues are divergent in the glucagon and GLP-1 amino termini; Ser2, Gln3, Tyr10, and Lys12 in glucagon and the corresponding Ala8, Glu9, Val16, and Ser18 in GLP-1. In this study, individual substitution of these four residues of glucagon with the corresponding residues of GLP-1 decreased the affinity and potency at the glucagon receptor relative to glucagon. Substitution of distinct segments of the glucagon receptor core domain with the corresponding segments of the GLP-1 receptor rescued the affinity and potency of specific glucagon analogs. Site-directed mutagenesis identified the Asp385 --> Glu glucagon receptor mutant that specifically rescued Ala2-glucagon. The results show that three distinct epitopes of the glucagon receptor core domain determine specificity for the N terminus of glucagon. We suggest a glucagon receptor binding model in which the extracellular ends of TM2 and TM7 are close to and determine specificity for Gln3 and Ser2 of glucagon, respectively. Furthermore, the second extracellular loop and/or proximal segments of TM4 and/or TM5 are close to and determine specificity for Lys12 of glucagon. PMID- 12724332 TI - Characterization of insulin inhibition of transactivation by a C-terminal fragment of the forkhead transcription factor Foxo1 in rat hepatoma cells. AB - The transcription factor Foxo1 controls the expression of genes involved in fundamental cellular processes. In keeping with its important physiological roles, Foxo1 activity is negatively regulated in response to growth factors and cytokines that activate a phosphatidylinositol 3-kinase (PI 3-kinase) protein kinase B (PKB)/Akt pathway. PKB/Akt-mediated phosphorylation of Foxo1 has been shown to result in the inhibition of target gene transcription and to trigger the export of Foxo1 from the nucleus, which is generally believed to explain the subsequent decrease of transcription. In the present study, using a chimeric protein in which a C-terminal fragment of Foxo1 (amino acids 208-652) containing the transactivation domain is fused to the yeast Gal4 DNA binding domain, we present evidence showing that insulin can directly regulate transactivation by Foxo1 in H4IIE rat hepatoma cells. Insulin inhibition of Foxo1-(208-652) stimulated transactivation is mediated by PI 3-kinase but in contrast to full length Foxo1, does not require either of the two PKB/Akt phosphorylation sites (Ser253 and Ser316) present in the protein fragment. Using mutational and deletion studies, we identify two potential phosphorylation sites, Ser319 and Ser499, as well as a 15-amino acid region located between residues 350 and 364 that are critical for insulin inhibition of transactivation by Foxo1-(208-652). We conclude that the transcriptional activity of Foxo1 is regulated at different levels by insulin: transactivation, as well as DNA binding and nuclear exclusion. These different regulatory mechanisms allow the precise control of transcription of Foxo1 target genes by insulin. PMID- 12724334 TI - Genetic research: mining for medical treasures. PMID- 12724335 TI - The discovery of captopril. PMID- 12724336 TI - Regulation of apoptosis: the ubiquitous way. AB - Ubiquitin is a ubiquitously expressed 76 amino acid protein that can be covalently attached to target proteins, leading to their ubiquitination. Many ubiquitinated proteins are degraded by the proteasome, a 2000 kDa ATP-dependent proteolytic complex. Numerous studies have demonstrated that the ubiquitination and proteasome system plays an important role in controlling the levels of various cellular proteins and therefore regulates basic cellular processes such as cell cycle progression, signal transduction, and cell transformation. Ubiquitination also directly affects the function and location of target proteins. Recent studies found that ubiquitination-mediated degradation and change in activity regulate many molecules of the cell death machinery, such as p53, caspases, and Bcl-2 family members. Ring finger-containing members of the IAP (inhibitor of apoptosis) family proteins themselves can function as ubiquitin protein ligases to ubiquitinate their target proteins or promote autoubiquitination. It has been demonstrated that degradation of the IAP proteins is required for apoptosis to occur in some systems, indicating apoptosis proceeds by activating death pathways as well as eliminating "roadblocks" through ubiquitination. These new findings also suggest that ubiquitination is one of the major mechanisms that regulate apoptotic cell death and could be a unique target for therapeutic intervention. PMID- 12724337 TI - Does the release of arachidonic acid from cells play a role in cancer chemoprevention? AB - COX-2 is overexpressed in cancer cells and has become a major target for cancer preventive drugs. NSAIDs, retinoids, antioxidants, and PPAR agonists, reported to be chemopreventive, suppress COX-2 synthesis. NSAIDs also have been shown to be chemopreventive independent of COX-2 activity. Common to all of these compounds is their ability to release arachidonic acid (AA) from rat liver cells in culture. Most of these compounds inhibit induced PGI2 production. Vitamin D3 and tamoxifen, however, not only stimulate the release of AA from cells: they amplify rather than inhibits induced COX activity. In view of the many activities attributable to AA, I propose that its release and accumulation could initiate molecular reactions that lead to apoptosis and eventually to suppression of cancer. Some drugs shown to release AA from cells and affect PGI2 production e.g., thiazolidinediones and statins are widely used for conditions unrelated to cancer. In vivo studies could reveal whether they can also function as cancer preventive agents. PMID- 12724338 TI - 4'-iodo-4'-deoxydoxorubicin and tetracyclines disrupt transthyretin amyloid fibrils in vitro producing noncytotoxic species: screening for TTR fibril disrupters. AB - Transthyretin Leu55Pro is one of the most aggressive mutations in familial amyloidotic polyneuropathy, an autosomal dominant disorder characterized by extracellular deposition of fibrillar amyloid protein. This variant has the ability to form fibrils in vitro under physiological conditions (PBS, pH 7.4). We studied by transmission electron microscopy the effect of the drug 4'-iodo-4' deoxydoxorubicin (I-DOX) on the in vitro assembly of TTR Leu55Pro fibrils by following fibril growth over a 15 day period. Our results showed that I-DOX at a concentration of 10-5 M/100 microg fibrils does not inhibit fibril formation in up to 10 days since fibrils identical to the ones present in the untreated sample were observed. However, after 15 days of treatment, only round particles, resembling soluble native TTR, were observed. We also tested the ability of tetracyclines and nitrophenols to interfere with amyloid fibril formation for 17 days; the group of compounds tested showed fibril disruption activity to different extents: doxycycline and 2,4-dinitrophenol resulted in complete disaggregation of fibrils. The species generated upon I-DOX and tetracyclines treatments were nontoxic, as revealed by the lack of significant caspase-3 activation on a Schwannoma cell line, making them potential therapeutic drugs in TTR-related and other amyloidosis. PMID- 12724339 TI - Evidence for an endogenous per1- and ICER-independent seasonal timer in the hamster pituitary gland. AB - Most mammals use changing annual day-length cycles to regulate pineal melatonin secretion and thereby drive many physiological rhythms including reproduction, metabolism, immune function, and pelage. Prolonged exposure to short winter day lengths results in refractoriness, a spontaneous reversion to long-day physiological status. Despite its critical role in the timing of seasonal rhythms, refractoriness remains poorly understood. The aim of this study was therefore to describe cellular and molecular mechanisms driving the seasonal secretion of a key hormone, prolactin, in refractory Syrian hamsters. We used recently developed single cell hybridization and reporter assays to show that this process is initiated by timed reactivation of endocrine signaling from the pars tuberalis (PT) region of the pituitary gland, a well-defined melatonin target site, causing renewed activation of prolactin gene expression. This timed signaling is independent of per1 clock gene expression in the suprachiasmatic nuclei and PT and of melatonin secretion, which continue to track day length. Within the PT, there is also a continued short day-like profile of ICER expression, suggesting that the change in hormone secretion is independent of cAMP signaling. Our data thus identify the PT as a key anatomical structure involved in endogenous seasonal timing mechanisms, which breaks from prevailing day length-induced gene expression. PMID- 12724340 TI - Gamma-tocopherol, but not alpha-tocopherol, decreases proinflammatory eicosanoids and inflammation damage in rats. AB - Gamma-tocopherol (gammaT), the major form of vitamin E in U.S. diets, and its physiological metabolite 2, 7, 8-trimethyl-2-(beta-carboxyethyl)-6-hydroxychroman (gamma-CEHC), in contrast to alpha-tocopherol (alphaT), the primary vitamin E in supplements, inhibit cyclooxygenase-catalyzed synthesis of prostaglandin E2 (PGE2) in activated macrophages and epithelial cells. Here we report that in carrageenan-induced inflammation in male Wistar rats, administration of gammaT (33 or 100 mg/kg) and gamma-CEHC (2 mg/pouch), but not alphaT (33 mg/kg), significantly reduced PGE2 synthesis at the site of inflammation. gammaT, but not alphaT, significantly inhibited the formation of leukotriene B4, a potent chemotactic agent synthesized by the 5-lipoxygenase of neutrophils. Although gammaT had no effect on neutrophil infiltration, it significantly attenuated the partial loss of food consumption caused by inflammation-associated discomfort. Administration of gammaT led consistently to a significant reduction of inflammation-mediated increase in 8-isoprostane, a biomarker of lipid peroxidation. gammaT at 100 mg/kg reduced TNF-alpha (65%;P=0.069), total nitrate/nitrite (40%;P=0.1), and lactate dehydrogenase activity (30%;P=0.067). Collectively, gammaT inhibits proinflammatory PGE2 and LTB4, decreases TNF-alpha, and attenuates inflammation-mediated damage. These findings provide strong evidence that gammaT shows anti-inflammatory activities in vivo that may be important for human disease prevention and therapy. PMID- 12724341 TI - A composite tissue-engineered trachea using sheep nasal chondrocyte and epithelial cells. AB - This study evaluates the feasibility of producing a composite engineered tracheal equivalent composed of cylindrical cartilaginous structures with lumens lined with nasal epithelial cells. Chondrocytes and epithelial cells isolated from sheep nasal septum were cultured in Ham's F12 media. After 2 wk, chondrocyte suspensions were seeded onto a matrix of polyglycolic acid. Cell-polymer constructs were wrapped around silicon tubes and cultured in vitro for 1 wk, followed by implanting into subcutaneous pockets on the backs of nude mice. After 6 wk, epithelial cells were suspended in a hydrogel and injected into the embedded cartilaginous cylinders following removal of the silicon tube. Implants were harvested 4 wk later and analyzed. The morphology of implants resembles that of native sheep trachea. H&E staining shows the presence of mature cartilage and formation of a pseudo-stratified columnar epithelium, with a distinct interface between tissue-engineered cartilage and epithelium. Safranin-O staining shows that tissue-engineered cartilage is organized into lobules with round, angular lacunae, each containing a single chondrocyte. Proteoglycan and hydroxyproline contents are similar to native cartilage. This study demonstrates the feasibility of recreating the cartilage and epithelial portion of the trachea using tissue harvested in a single procedure. This has the potential to facilitate an autologous repair of segmental tracheal defects. PMID- 12724342 TI - Direct gene transfer into rat articular cartilage by in vivo electroporation. AB - To establish a system for efficient direct in vivo gene targeting into rat joint, we have evaluated a strategy of gene transfer by means of the delivery of external electric pulses (EP) to the knee after intra-articular injection of a reporter gene (GFP). Rats were killed at various times after the electro gene therapy to analyze GFP gene expression by immunohistochemistry. GFP staining was detected in the superficial, middle, and deep zones of the patellar cartilage at days 2 and 9, and thereafter only in the deep zone (months 1 and 2). The average percentage of GFP-positive cells was estimated at 30% both one and 2 months after the gene transfer. Moreover, no pathologic change caused by the EP was detected in the cartilage. The level and stability of the long-term GFP expression found in this study demonstrate the feasibility of a treatment of joint disorders (inflammatory or degenerative, focal or diffuse) using electric gene transfer. PMID- 12724343 TI - Cascades of transcriptional induction during dendritic cell maturation revealed by genome-wide expression analysis. AB - Dendritic cells (DC) are central regulators of immunity. Signal-induced maturation of DCs is assumed to be the starting point for specific immune responses. To further understand this process, we analyzed the alteration of transcript profiles along the time course of CD40 ligand-induced maturation of human myeloid DCs by Affymetrix GeneChip microarrays covering >6800 genes. Besides rediscovery of genes already described as associated with DC maturation proving reliability of the methods used, we identified clusterin as novel maturation marker. Looking across the time course, we observed synchronized kinetics of distinct functional groups of molecules whose temporal coregulation underscores known cellular events during dendritic cell maturation. For example, an early-peaking wave of inflammatory chemokines was followed by a sustained increase of constitutive chemokines and accompanied by slow but continuous induction of survival proteins. After an immediate but transient induction of cytokine-responsive transcripts, there was an increased expression of a group of genes involved in not only the regulation of cytokine effects, but also of transcription in general. Our results demonstrate that microarray studies along time courses combined with real-time PCR not only discover new marker molecules with functional implications, but also dissect the molecular kinetics of biological processes identifying complex pathways of regulation. PMID- 12724344 TI - Heterotrimeric G-proteins associate with microtubules during differentiation in PC12 pheochromocytoma cells. AB - Tubulin modifies G-protein signaling and heterotrimeric G-proteins regulate microtubule assembly. Here we report an interplay among G-protein-coupled receptor and receptor tyrosine kinase (such as nerve growth factor-NGF) signaling systems in PC12 pheochromocytoma cells that resulted in a translocation of Galpha(s), Galpha(i1), and Galpha(o) from cell bodies to cellular processes where they appear to localize with tubulin-containing structures. This relocation appeared to depend on the integrity of microtubules, as it was blocked and reversed by nocodazole. Latrunculin, which promotes actin filament depolymerization, had no effect. Both deconvolution microscopy and immunoprecipitation showed a significant increase of Galpha association with microtubules that was coincident with the extension of "neurites." There were distinctions among the Galpha subtypes, with Galpha(s) showing the most profound NGF-induced colocalization with tubulin. Translocation of Galpha was blocked by agents that inhibit the MAP kinases required for neuronal differentiation, suggesting that G-protein relocation is triggered by the intracellular signals for differentiation. Consistent with this, Galpha in Neuro-2A cells, which spontaneously differentiate, showed a similar translocation coincident with differentiation. Thus, diverse signals that promote neuronal differentiation and changes in cell morphology may use specific G-proteins to evoke cytoskeletal rearrangement. PMID- 12724345 TI - The importance of cell-mediated immunity in the course and severity of autoimmune anti-glomerular basement membrane disease in mice. AB - Anti-glomerular basement membrane (GBM) disease is a rapidly progressive glomerulonephritis (GN) resulting from autoimmunity against the Goodpasture antigen alpha3(IV)NC1. In addition to the well-characterized antibody contribution, a T helper 1 (Th1) response has been suspected as the culprit for glomerular injury. We induced anti-GBM disease in DBA/1, C57BL/6, AKR, and NOD mice with recombinant human alpha3(IV)NC1 to investigate the involvement of humoral and cellular autoimmunity. DBA/1 mice had crescentic GN 11 wk postimmunization with alpha3(IV)NC1. C57BL/6 and AKR mice developed a chronic disease course resulting in comparable kidney injury to DBA/1 mice within 6 months. NOD revealed only minor glomerular changes. The rapid course and the severity of the disease in DBA/1 mice can be explained by our immunological findings in their sera and splenocytes: 1) high antibody titers specific for the putative clinically relevant epitope of alpha3(IV)NC1 with Th1-type isotypes, and 2) a strong proliferative response and high amounts of the inflammatory cytokine IFN-gamma, secreted by splenocytes stimulated in vitro with alpha3(IV)NC1, with only low amounts of the anti-inflammatory cytokine IL-10. Our in vivo and in vitro results provide direct evidence that the balance between Th1 and Th2 responses associates with the outcome of anti-GBM disease in mice. PMID- 12724348 TI - The expression of delta- and kappa-opioid receptor is enhanced during intestinal inflammation in mice. AB - In the gut, mu-, delta-, and kappa-opioid receptors are present in the submucous and myenteric plexi and in enterocytes. Using pharmacological methods, our group has shown that intestinal inflammation enhances the antitransit and antisecretory effects of systemic opioids. The aim of the present study was to evaluate whether the enhanced antisecretory effects of delta and kappa-agonists were associated with an increased transcription and/or expression of these receptors at central (brain and spinal cord) and/or peripheral sites (gut); we also evaluated the expression of delta- and kappa-opioid receptors in dissected sections of the gut containing the myenteric (MP/LM) or submucous (SP/M) plexi. The mRNA and protein levels of both opioid receptors were determined using a reverse-transcriptase polymerase chain reaction and immunoprecipitation/Western blot, respectively. Intestinal inflammation significantly augmented the transcription of delta-opioid receptors in the spinal cord (34%) and in the whole gut (102%). Also increased mRNA and protein levels of delta-opioid receptors in the MP/LM and SP/M preparations. The kappa-opioid receptors gene transcription was not altered by inflammation, whereas kappa-opioid receptors protein levels were significantly enhanced in the SP/M preparation. No changes in gene transcription or protein levels for delta- and kappa-opioid receptors could be demonstrated in the brain. These results suggest that local transcriptional and post-transcriptional changes of the delta- and kappa-opioid receptors genes could be responsible for the enhanced antisecretory effects of delta- and kappa-opioid agonists during intestinal inflammation. PMID- 12724347 TI - Stimulatory action of itopride hydrochloride on colonic motor activity in vitro and in vivo. AB - We investigated the effects of itopride hydrochloride (itopride, N-[4-[2 (dimethylamino)ethoxy]benzyl]-3,4-dimethoxybenzamide hydrochloride), a gastroprokinetic agent, on the colonic motor activity in vitro and in vivo, in comparison with benzamides, cisapride hydrate (cisapride), and mosapride citrate (mosapride). Itopride stimulated both peristaltic and segmental motility induced by applying intraluminal pressure to the isolated guinea pig colon. Although cisapride and mosapride enhanced the segmental motility, they markedly reduced the peristaltic motility. In conscious dogs with implanted strain gauge force transducers, itopride stimulated contractile activity in the gastrointestinal tract from the stomach to the colon. Cisapride stimulated contractile activity in the gastric antrum, ileum, and ascending colon. Mosapride stimulated contractile activity only in the gastric antrum and ileum. In guinea pigs and rats, itopride accelerated colonic luminal transit. On the other hand, cisapride and mosapride failed to enhance colonic transit. These results demonstrate that itopride has a stimulatory action on colonic peristalsis, propelling colonic luminal contents, different from that of cisapride and mosapride. Therefore, itopride may be a useful drug for the treatment of functional bowel disorders such as functional constipation. PMID- 12724346 TI - Receptor-coupled, DAG-gated Ca2+-permeable cationic channels in LNCaP human prostate cancer epithelial cells. AB - Although the prostate gland is a rich source of alpha1-adreno- (alpha1-AR) and m1 cholino receptors (m1-AChR), the membrane processes associated with their activation in glandular epithelial cells is poorly understood. We used the whole cell patch-clamp technique to show that the agonists of the respective receptors, phenylephrine (PHE) and carbachol (CCh), activate cationic membrane currents in lymph node carcinoma of the prostate (LNCaP) human prostate cancer epithelial cells, which are not dependent on the filling status of intracellular IP3 sensitive Ca2+ stores, but directly gated by diacylglycerol (DAG), as evidenced by the ability of its membrane permeable analogue, OAG, to mimic the effects of the agonists. The underlying cationic channels are characterized by the weak field-strength Eisenman IV permeability sequence for monovalent cations (PK(25) > PCs(4.6) > PLi(1.4) > PNa(1.0)), and the following permeability sequence for divalent cations: PCa(1.0) > PMg(0.74) > PBa(0.6) > PSr(0.36) > PMn(0.3). They are 4.3 times more permeable to Ca2+ than Na+ and more sensitive to the inhibitor 2-APB than SK&F 96365. RT-PCR analysis shows that DAG-gated members of the transient receptor potential (TRP) channel family, including TRPC1 and TRPC3, are present in LNCaP cells. We conclude that, in prostate cancer epithelial cells, alpha1-ARs and m1-AChRs are functionally coupled to Ca2+-permeable DAG-gated cationic channels, for which TRPC1 and TRPC3 are the most likely candidates. PMID- 12724349 TI - Mutational uncoupling of alpha1A-adrenergic receptors from G proteins also uncouples mitogenic and transcriptional responses in PC12 cells. AB - Activation of human alpha1A-adrenergic receptors in PC12 cells causes many second messenger, mitogenic, and transcriptional responses. We examined the role of G protein activation in these responses by uncoupling the receptor through deletion of the first three amino acids from the third intracellular loop (Delta208-210). Expression levels of retrovirus-transfected wild-type and Delta(208-210) alpha1A adrenergic receptors in PC12 cells were similar and showed identical binding affinities for antagonists. However, the potency of the agonist norepinephrine was increased 9-fold by the Delta (208-210) mutation. In PC12 cells expressing the Delta (208-210) construct, calcium and inositol phosphate responses to norepinephrine were essentially abolished. The strong activation of mitogen activated protein kinase pathways seen upon stimulation of wild-type alpha1A adrenergic receptors in PC12 cells was abolished by the Delta (208-210) mutation, as was activation of the tyrosine kinase Pyk2. Norepinephrine also activates several transcriptional reporters through alpha1A-adrenergic receptors in PC12 cells, including reporters for activator protein 1, serum response element, cAMP response element, nuclear factor-kappaB, nuclear factor of activated T cells, gamma-interferon-activated sequence, and signal transducer and activator of transcription. All these transcriptional responses were abolished by the Delta (208-210) mutation. Overexpression of Galpha16 did not rescue any of these responses. These data suggest that known second messenger, mitogenic, and transcriptional effects of alpha1A-adrenergic receptors in PC12 cells all require G protein activation. PMID- 12724350 TI - Increased expression of vanilloid receptor 1 on myelinated primary afferent neurons contributes to the antihyperalgesic effect of capsaicin cream in diabetic neuropathic pain in mice. AB - Topical capsaicin is believed to alleviate pain by desensitizing the vanilloid receptor 1 (VR1) at the peripheral nerve endings. Here, we report that an up regulation of VR1 expression on myelinated fibers contributes to the antihyperalgesic effect of capsaicin cream in streptozotocin (STZ)-induced diabetic neuropathic pain. Intravenous injection of STZ (200 mg/kg) in mice caused rapid onset of diabetes within 24 h. Thermal and mechanical hyperalgesia developed by 3 days after STZ injection and persisted at all time points tested until 28 days. There was also hyperalgesic response to intraplantar (i.pl.) prostaglandin I2 (PGI2) agonist-induced nociception in such mice. Application of capsaicin cream dose dependently reversed the thermal, mechanical, and PGI2 agonist-induced hyperalgesia observed in the diabetic mice. The i.pl. injection of capsaicin solution (0.4 microg/20 microl) produced nociceptive biting-licking responses in control mice, and these responses were significantly increased in STZ-induced diabetic mice. After neonatal capsaicin-treatment, which destroys most unmyelinated C-fibers, the i.pl. capsaicin-induced biting-licking responses were almost abolished. However, in neonatal capsaicin-treated diabetic mice, the i.pl. capsaicin-induced biting-licking responses reappeared. The i.pl. capsaicin induced biting-licking responses were blocked by the competitive VR1 antagonist capsazepine. All these results suggest an increase in capsaicin receptor on myelinated fibers due to diabetes. Finally, we confirmed the up-regulation of VR1 expression on myelinated primary afferent neurons of diabetic mice by immunohistochemistry. Together, our results suggest that increased expression of VR1 on myelinated fibers might contribute to the antihyperalgesic effect of topical capsaicin in diabetic neuropathic pain. PMID- 12724351 TI - Involvement of human organic anion transporting polypeptide OATP-B (SLC21A9) in pH-dependent transport across intestinal apical membrane. AB - Some organic anions are absorbed from the gastrointestinal tract through carrier mediated transport mechanism(s), which may include proton-coupled transport, anion exchange transport, and others. However, the molecular identity of the organic anion transporters localized at the apical membrane of human intestinal epithelial cells has not been clearly demonstrated. In the present study, we focused on human organic anion transporting polypeptide OATP-B and examined its subcellular localization and functionality in the small intestine. Localization of OATP-B was determined by immunohistochemical analysis. Transport properties of estrone-3-sulfate and the 3-hydroxy-3-methylglutaryl-CoA reductase inhibitor pravastatin by OATP-B-transfected human embryonic kidney 293 cells were measured. OATP-B was immunohistochemically localized at the apical membrane of intestinal epithelial cells in humans. Uptake of [3H]estrone-3-sulfate and [14C]pravastatin by OATP-B at pH 5.5 was higher than that at pH 7.4. [3H]Estrone-3-sulfate transport was decreased by pravastatin, aromatic anion compounds, and the anion exchange inhibitor 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid, but not by small anionic compounds, such as lactic acid and acetic acid. The inhibitory effect of pravastatin on the uptake of [3H]estrone-3-sulfate was concentration dependent, and the IC50 value was 5.5 mM. The results suggested that OATP-B mediates absorption of anionic compounds and its activity may be optimum at the acidic surface microclimate pH of the small intestine. Accordingly, OATP-B plays a role in the absorption of anionic compounds across the apical membrane of human intestinal epithelial cells, although it cannot be decisively concluded that pH dependent absorption of pravastatin is determined by OATP-B alone. PMID- 12724352 TI - Drosophila securin destruction involves a D-box and a KEN-box and promotes anaphase in parallel with Cyclin A degradation. AB - Sister chromatid separation during exit from mitosis requires separase. Securin inhibits separase during the cell cycle until metaphase when it is degraded by the anaphase-promoting complex/cyclosome (APC/C). In Drosophila, sister chromatid separation proceeds even in the presence of stabilized securin with mutations in its D-box, a motif known to mediate recruitment to the APC/C. Alternative pathways might therefore regulate separase and sister chromatid separation apart from proteolysis of the Drosophila securin PIM. Consistent with this proposal and with results from yeast and vertebrates, we show here that the effects of stabilized securin with mutations in the D-box are enhanced in vivo by reduced Polo kinase function or by mitotically stabilized Cyclin A. However, we also show that PIM contains a KEN-box, which is required for mitotic degradation in addition to the D-box, and that sister chromatid separation is completely inhibited by PIM with mutations in both degradation signals. PMID- 12724353 TI - WIP participates in actin reorganization and ruffle formation induced by PDGF. AB - Platelet-derived growth factor (PDGF) is a chemotactic factor for fibroblasts that triggers actin cytoskeleton reorganization by increasing the level of GTP Rac, the activated form of a small Rho family GTPase. GTP-Rac induces membrane ruffling and lamellipodium formation that are required for adhesion, migration and macropinocytosis, among other functions. We have shown that WIP interacts with members of the Wiskott-Aldrich syndrome protein family and is essential for filopodium formation regulated by Cdc42 GTPase. In this report, we show that WIP participates in the actin reorganization that leads to ruffle formation. WIP overexpression in murine fibroblasts (3T3 cells) enhances ruffle formation in response to PDGF stimulation, as shown by immunofluorescence and electron and video microscopy. More importantly, microinjection of anti-WIP antibody or absence of WIP in murine fibroblasts results in decreased ruffle formation in response to PDGF treatment. Finally, overexpression of a modified form of WIP lacking the actin-binding site blocks PDGF-induced membrane ruffling. These data suggest a role for WIP in actin reorganization to form PDGF-induced ruffles. This is the first in vivo evidence in mammalian cells for a function of WIP dependent on its ability to bind actin. PMID- 12724354 TI - Thrombin induces endothelial cell-surface exposure of the plasminogen receptor annexin 2. AB - Cell-surface annexin 2 (A2) and its ligand p11 have been implicated in fibrinolysis because of their ability to accelerate tissue plasminogen activator (tPA)-mediated activation of plasminogen to plasmin. Because thrombin is a potent cell modulator obligately produced at the site of clot formation, we hypothesized that the amount of cell-surface A2 and p11 might be altered by thrombin with consequent effects on plasmin generation. In support of this hypothesis, immunofluorescence microscopy and hydrophilic biotinylation experiments showed that both A2 and p11 were significantly increased on the surface of human umbilical vein endothelial cells (HUVECs) treated with thrombin (0.8-8 nM) for 5 minutes followed by 1 hour at 37 degrees C. Intracellular immunofluorescence microscopy and immunoblot analyses of whole cell extracts revealed increased p11 but unchanged A2 in response to thrombin, suggesting that transbilayer trafficking of A2 might be controlled by p11. The thrombin receptor-activating peptide (TRAP) similarly affected cells, demonstrating that cell signaling at least involved the type-1 protease activated receptor (PAR-1). An effect on the fibrinolysis pathway after treatment of HUVECs with thrombin was shown by increased fluorescein-labeled plasminogen binding to cells, which was inhibited by an antibody specific for p11. This was confirmed by observing that thrombin pretreatment of HUVECs increased biotin-modified plasminogen binding. Utilizing a chromogenic assay, pretreatment of HUVECs by thrombin further enhanced activation of the Glu and Lys forms of plasminogen by tPA. These data suggest a novel mechanism that links the coagulation and fibrinolysis pathways by thrombin mediated feedback. PMID- 12724355 TI - SFRP1 modulates retina cell differentiation through a beta-catenin-independent mechanism. AB - Secreted frizzled related proteins (SFRPs) are soluble molecules capable of binding WNTS and preventing the activation of their canonical signalling cascade. Here we show that Sfrp1 contributes to chick retina differentiation with a mechanism that does not involve modifications in the transcriptional activity of beta-catenin. Thus, addition of SFRP1 to dissociated retinal cultures or retroviral mediated overexpression of the molecule consistently promoted retinal ganglion and cone photoreceptor cell generation, while decreasing the number of amacrine cells. Measure of the activity of the beta-catenin-responsive Tcf binding site coupled to a luciferase reporter in transiently transfected retinal cells showed that Sfrp1 was unable to modify the basal beta-catenin transcriptional activity of the retina cells. Interestingly, a dominant-negative form of GSK3beta gave similar results to those of Sfrp1, and a phosphorylation dependent inhibition of GSK3beta activity followed SFRP1 treatment of retina cells. Furthermore, retroviral mediated expression of a dominant-negative form of GSK3beta induced a retina phenotype similar to that observed after Sfrp1 overexpression, suggesting a possible involvement of this kinase in SFRP1 function. PMID- 12724356 TI - Biogenesis and nuclear export of ribosomal subunits in higher eukaryotes depend on the CRM1 export pathway. AB - The production of ribosomes constitutes a major biosynthetic task for cells. Eukaryotic small and large ribosomal subunits are assembled in the nucleolus and independently exported to the cytoplasm. Most nuclear export pathways require RanGTP-binding export receptors. We analyzed the role of CRM1, the export receptor for leucine-rich nuclear export signals (NES), in the biogenesis of ribosomal subunits in vertebrate cells. Inhibition of the CRM1 export pathway led to a defect in nuclear export of both 40S and 60S subunits in HeLa cells. Moreover, the export of newly made ribosomal subunits in Xenopus oocytes was efficiently and specifically competed by BSA-NES conjugates. The CRM1 dependence of 60S subunit export suggested a conserved function for NMD3, a factor proposed to be a 60S subunit export adaptor in yeast. Indeed, we observed that nuclear export of human NMD3 (hNMD3) is sensitive to leptomycin B (LMB), which inactivates CRM1. It had, however, not yet been demonstrated that Nmd3 can interact with CRM1. Using purified recombinant proteins we have shown here that hNMD3 binds to CRM1 directly, in a RanGTP-dependent manner, by way of a C terminal NES sequence. Our results suggest that the functions of CRM1 and NMD3 in ribosomal subunit export are conserved from yeast to higher eukaryotes. PMID- 12724357 TI - PTHrP [67-86] regulates the expression of stress proteins in breast cancer cells inducing modifications in urokinase-plasminogen activator and MMP-1 expression. AB - It was previously reported that a midregion domain of parathyroid hormone-related protein (PTHrP), that is, [67-86]-amide, is able to restrain growth and promote matrigel penetration by the 8701-BC cell line, derived from a biopsy fragment of a primary ductal infiltrating carcinoma of the human breast, and that cell invasion in vitro is drastically impaired by inactivation of urokinase plasminogen activator (uPa). In this study we started a more detailed investigation of the possible effects on gene expression arising from the interaction between PTHrP [67-86]-amide and 8701-BC breast cancer cells by a combination of conventional-, differential display-and semi-quantitative multiplex-polymerase chain reaction (PCR) assays. We present here the first evidence that the upregulation of some stress-related genes, most noticeably heat shock factor binding protein-1 (hsbp1) and heat shock protein 90 (hsp-90), is involved in the acquisition of an in vitro more invasive phenotype by cells treated with midregion PTHrP. This is conceivably accomplished by sequestering and inactivating heat shock factor-1 (hsf1) which is able to recognize Ets transcription-factor-binding sites present in some gene promoters, such as those of uPa and matrix metalloprotease-1 (MMP-1). In fact, our data show that incubation of PTHrP [67-86]-amide-treated cells with either antisense hsbp1 oligonucleotide or geldanamycin, an hsp90-inactivating antibiotic, results in downregulation of uPa and upregulation of MMP-1, and in a prominent inhibition of cell invasion in matrigel-containing Transwell chambers. Alternatively, incubation of untreated 8701-BC cells with quercetin, a flavonoid known to decrease the amount of free hsf1, is found to induce upregulation of uPa and downregulation of MMP-1, and an increase of matrigel invasion by cells, thus providing further supporting data of the involvement of hsf unavailability on the modulation of uPa and MMP-1 expression and on cell invasive behaviour. These studies confirm a previous postulate that over-secretion of uPa, rather than of other extracellular proteases, is a primary condition for the increase of invasive activity triggered by PTHrP [67-86]-amide in vitro, and support a role for midregion forms of PTHrP in potentially affecting pathological mammary growth and differentiation. They also identify two new key protagonists in the complex scenario of breast tumor cell invasiveness in vitro, that is, hsbp1 and hsp90, which deserve further and more extensive studies as potential and attractive molecular targets for anti-breast cancer treatments. PMID- 12724358 TI - Local signals from beyond the receptive fields of striate cortical neurons. AB - We examined in anesthetized macaque how the responses of a striate cortical neuron to patterns inside the receptive field were altered by surrounding patterns outside it. The changes in a neuron's response brought about by a surround are immediate and transient: they arise with the same latency as the response to a stimulus within the receptive field (this argues for a source locally in striate cortex) and become less effective as soon as 27 ms later. Surround signals appeared to exert their influence through divisive interaction (normalization) with those arising in the receptive field. Surrounding patterns presented at orientations slightly oblique to the preferred orientation consistently deformed orientation tuning curves of complex (but not simple) cells, repelling the preferred orientation but without decreasing the discriminability of the preferred grating and ones at slightly oblique orientations. By reducing responsivity and changing the tuning of complex cells locally in stimulus space, surrounding patterns reduce the correlations among responses of neurons to a particular stimulus, thus reducing the redundancy of image representation. PMID- 12724359 TI - Antagonistic modulation of a hyperpolarization-activated Cl(-) current in Aplysia sensory neurons by SCP(B) and FMRFamide. AB - Whole cell voltage-clamp recordings from Aplysia mechanosensory neurons obtained from the pleural ganglion were used to investigate the actions on membrane currents of the neuropeptides SCP(B) and FMRFamide. At the start of whole cell recording, SCP(B) typically evoked an inward current at a holding potential of 40 mV, due to the cAMP-mediated closure of the S-type K+ channel, whereas FMRFamide evoked an outward current, due to the opening of the S-type K+ channels mediated by 12-lipoxygenase metabolites of arachidonic acid. However, after several minutes of whole cell recording with a high concentration of chloride in the whole cell patch pipette solution, the responses to SCP(B) and FMRF-amide at 40 mV were inverted; SCP(B) evoked an outward current, whereas FMRFamide and YGGFMRFamide evoked inward currents. Ion substitution experiments and reversal potential measurements revealed that these responses were due to the opposing regulation of a Cl(-) current, whose magnitude was greatly enhanced by dialysis with the high Cl(-) - containing pipette solution. SCP(B) inhibited this Cl(-) current through production of cAMP and activation of PKA. YGGFMRFamide activated this Cl(-) current by stimulating a cGMP-activated phosphodiesterase that hydrolyzed cAMP. Thus a cAMP-dependent Cl(-) current undergoes antagonistic modulation by two neuropeptides in Aplysia sensory neurons. PMID- 12724360 TI - Metabotropic receptor-mediated Ca2+ signaling elevates mitochondrial Ca2+ and stimulates oxidative metabolism in hippocampal slice cultures. AB - Metabotropic receptors modulate numerous cellular processes by intracellular Ca2+ signaling, but less is known about their role in regulating mitochondrial metabolic function within the CNS. In this study, we demonstrate in area CA3 of rat organotypic hippocampal slice cultures that glutamatergic, serotonergic, and muscarinic metabotropic receptor ligands, namely trans-azetidine-2,4-dicarboxylic acid, alpha-methyl-5-hydroxytryptamine, and carbachol, transiently increase mitochondrial Ca2+ concentration ([Ca2+]m) as recorded by changes in Rhod-2 fluorescence, stimulate mitochondrial oxidative metabolism as revealed by elevations in NAD(P)H fluorescence, and induce K+ outward currents as monitored by rapid increases in extracellular K+ concentration ([K+]o). Carbachol (1-1,000 microM) elevated NAD(P)H fluorescence by VPM) of the rodent whisker-to-barrel pathway transforms afferent signals, we applied ramp-and hold deflections to individual whiskers of lightly narcotized rats while recording the extracellular responses of neurons in either the ventroposterior medial (VPM) thalamic nucleus or in brain stem nucleus principalis (PrV). In PrV, only those neurons antidromically determined to project to VPM were selected for recording. We found that VPM neurons exhibited smaller response magnitudes and greater spontaneous firing rates than those of their PrV inputs, but that both populations were similarly well tuned for stimulus direction. In addition, fewer VPM (74%) than PrV neurons (93%) responded with sustained, or tonic, discharges during the plateau phase of the stimulus. Neurons in both populations responded most robustly to deflections of a single, "principal whisker" (PW), and the majority of cells in both PrV (90%) and VPM (73%) also responded to deflections of at least one adjacent whisker (AW). AW responses in both nuclei occurred on average at longer latencies and were more temporally dispersed than PW responses. Lateral inhibition, as evidenced by AW-evoked activity suppression, was rare in PrV but prevalent in VPM. In both nuclei, however, suppression was weak, with AW responses being on average excitatory. Our results suggest that the receptive field structures and response properties of individual VPM neurons can be explained in large part by input from one or a small number of PrV neurons, but that intrathalamic mechanisms act to further transform the afferent signal. PMID- 12724363 TI - Inflammation induces ectopic mechanical sensitivity in axons of nociceptors innervating deep tissues. AB - A variety of seemingly diverse pain syndromes are characterized by movement induced pain radiating in the distribution of a peripheral nerve or nerve root. This could be explained by the induction of ectopic mechanical sensitivity in intact sensory axons. Here we show that inflammation led to mechanical sensitivity of the axons of a subset of mechanically sensitive primary sensory neurons. Dorsal root recordings were made from 194 mechanically sensitive neurons that innervated deep and cutaneous structures and had C, Adelta, and Aalphabeta conduction velocities. No axons of any category were mechanically sensitive in control experiments. However, the axons of neurons innervating deep structures and having C- or Adelta-conduction velocities became mechanically sensitive during the neuritis, and also exhibited an increased incidence of spontaneous discharge. The incidence of mechanical sensitivity followed a distinct time course. In some cases, paw withdrawal thresholds were obtained after neuritis induction. The time course of the resultant hypersensitivity was not directly related to the time course of the axonal mechanical sensitivity. Ectopic axonal mechanical sensitivity could explain some types of radiating, nerve-related pain coexisting with diseases of seemingly diverse etiologies. PMID- 12724365 TI - Flash photolysis reveals a diversity of ionotropic glutamate receptors on the mitral cell somatodendritic membrane. AB - It is widely held that the soma and basal dendrites of olfactory bulb mitral cells receive exclusively inhibitory synaptic input from local interneurons. However, the mitral somatodendritic membrane exhibits immunoreactivity for a variety of glutamate receptors, and blocking GABA receptors unmasks mitral cell self-excitation. This excitation is proposed to be mediated either by diffuse spillover of the mitral cells' own released glutamate, or by punctate transmission from glutamate-releasing granule cells. This study examined the pharmacology and kinetics of glutamate sensitivity of mitral cells by flash photolysis of nitroindoline caged glutamates, which facilitate reliable activation of receptors in the synaptic cleft. Wide-field laser uncaging (3.5-ms flash) of approximately 0.5-1 mM glutamate onto the soma activated large currents with fast (3.4-ms rise, 7.5-ms decay) and slow (64-ms rise, >10-s decay) components. In 100 microM APV, slow currents were reduced to 53% of control (257 ms rise, 2-s decay), displayed outward rectification in 1.3 mM Mg2+, and blocked by 15 microM 5,7-dichlorokynurenate. Responses to less, similar 100 microM glutamate were fully antagonized by 100 microM APV, consistent with competitive inhibition at high-affinity NMDA receptors. An APV-resistant NMDA receptor was not observed, refuting the punctate transmission model. Fast currents were blocked by 10 microM NBQX, boosted 3.28-fold by 100 microM cyclothiazide, and resolved into AMPA (40%) and kainate (60%) receptor components by 100 microM SYM2206. The results suggest that self-excitation depends on AMPA, kainite, and conventional NMDA autoreceptors on the mitral cell. PMID- 12724366 TI - A minimal model for G protein-mediated synaptic facilitation and depression. AB - G protein-coupled receptors are ubiquitous in neurons, as well as other cell types. Activation of receptors by hormones or neurotransmitters splits the G protein heterotrimer into Galpha and Gbetagamma subunits. It is now clear that Gbetagamma directly inhibits Ca2+ channels, putting them into a reluctant state. The effects of Gbetagamma depend on the specific beta and gamma subunits present, as well as the beta subunit isoform of the N-type Ca2+ channel. We describe a minimal mathematical model for the effects of G protein action on the dynamics of synaptic transmission. The model is calibrated by data obtained by transfecting G protein and Ca2+ channel subunits into tsA-201 cells. We demonstrate with numerical simulations that G protein action can provide a mechanism for either short-term synaptic facilitation or depression, depending on the manner in which G protein-coupled receptors are activated. The G protein action performs high pass filtering of the presynaptic signal, with a filter cutoff that depends on the combination of G protein and Ca2+ channel subunits present. At stimulus frequencies above the cutoff, trains of single spikes are transmitted, while only doublets are transmitted at frequencies below the cutoff. Finally, we demonstrate that relief of G protein inhibition can contribute to paired-pulse facilitation. PMID- 12724368 TI - Nitric oxide differentially modulates ON and OFF responses of retinal ganglion cells. AB - Several lines of evidence suggest that nitric oxide (NO) can regulate diverse retinal functions, but whether this gas is capable of modulating the visual responses of retinal output neurons has not been established. In the present study the effects of NO on rod-driven responses of retinal ganglion cells were tested by making whole cell patch-clamp recordings from morphologically identified ganglion cells in the isolated ferret retina. Bath application of L arginine, the substrate of nitric oxide synthase, and S-nitroso-N acetylpenicillamine, the NO donor, was found to differentially affect on and off discharge patterns. The introduction of these drugs significantly decreased visual responses of retinal ganglion cells, but the effects were more pronounced on off than on on discharges. The peak discharge rates of on responses were usually reduced by about 40%, but not completely blocked. In contrast, off responses were completely blocked in most cells. These differential effects were observed in on-off cells as well as in cells that yielded just on or off discharges. The off responses that were blocked by NO were also blocked by DL-2 amino-phosphonobutyric acid (APB) and strychnine, suggesting the involvement of the APB-sensitive rod pathway. PMID- 12724369 TI - Reciprocal interactions between calcium and chloride in rod photoreceptors. AB - This study used imaging and electrophysiological techniques in salamander retinal slices to correlate Ca2+ and Cl- levels in rods and thus test the hypothesis of a feedback interaction between Ca2+- and Ca2+-activated Cl- channels whereby Cl- efflux through Cl- channels can inhibit Ca2+ channels. Increasing [K+]o levels produced a concentration-dependent depolarization of rods accompanied by increases in [Ca2+]i measured with Fura-2. The voltage dependence of increases in [Ca2+]i was compared with the voltage dependence of the calcium current (ICa). [Cl-]i was measured with the dye, MEQ. Depolarization with high K+ to membrane potentials below -20 mV reduced [Cl-]i; larger depolarizations increased [Cl-]i. The Na/K/Cl cotransport inhibitor, bumetanide, shifted the apparent Cl- equilibrium potential (ECl) to more negative potentials, suggesting that this cotransporter helps establish a relatively depolarized ECl. MEQ fluorescence changes evoked by high K+ were inhibited by niflumic acid (0.1 mM), NPPB (2 microM), or replacement of Ca2+ with Ba2+, suggesting that depolarization-evoked Cl- changes result partly from stimulation of Ca2+-activated Cl- channels. Replacing >/=12 mM [Cl-]o with CH3SO4- produced a significant reduction in [Cl ]i. [Ca2+]i increases evoked by 20 or 50 mM K+ were also significantly inhibited by replacing >/=12 mM [Cl-]o with CH3SO4-. Thus modest depolarization can evoke increases in [Ca2+]i that lead to reductions in [Cl-]i, and conversely, reductions in [Cl-]i inhibit depolarization-evoked [Ca2+]i increases. These findings support the hypothesis that feedback interactions between Ca2+- and Ca2+ activated Cl- channels may contribute to the regulation of presynaptic Ca2+ currents involved in synaptic transmission from rod photoreceptors. PMID- 12724367 TI - Persistent sodium and calcium currents cause plateau potentials in motoneurons of chronic spinal rats. AB - After chronic spinal cord injury motoneurons exhibit large plateau potentials (sustained depolarizations triggered by brief inputs) that play a primary role in the development of muscle spasms and spasticity (Bennett et al. 2001a,b). The present study examined the voltage-gated persistent inward currents (PICs) underlying these plateaus. Adult rats were spinalized at the S2 sacral spinal level and after 2 mo, when spasticity developed, intracellular recordings were made from motoneurons below the injury. For recording, the whole sacrocaudal spinal cord was removed and maintained in vitro in normal artificial cerebral spinal fluid (nACSF), without application of neuromodulators. During a slow triangular voltage-clamp command (ramp) a PIC was activated with a threshold of 54.2 +/- 4.8 mV (similar to plateau threshold), with a peak current of 2.88 +/- 0.95 nA and produced a pronounced negative-slope region in the V-I relation. This PIC was in part mediated by Cav1.3 L-type calcium channels because it was low threshold and significantly reduced by 10 to 20 microM nimodipine or 400 microM Cd2+. The PIC that remained during a calcium channel blockade (in Cd2+) was completely and rapidly blocked by tetrodotoxin (TTX; 0.5 to 2 microM), and thus was a TTX-sensitive persistent sodium current. This persistent sodium current was activated rapidly about 7 mV below the spike threshold (spike threshold -46.1 +/- 4.5 mV), contributed approximately 1/2 of the initial peak of the total PIC, inactivated partly to contribute only approximately 1/3 of the sustained PIC (at 5 to 10 s), and deactivated rapidly with hyperpolarization (<50 ms). When TTX was added to the bath first, the nimodipine-sensitive persistent calcium current (L type) was seen in isolation; it was slowly activated (>250 ms), had a low but variable threshold (either slightly above or below the spike threshold), contributed the other approximately 1/2 of the initial peak of the total PIC (before TTX), did not usually inactivate with time (contributed approximately two thirds of the sustained PIC), and deactivated slowly with hyperpolarization to rest (in >300 ms). In summary, low-threshold persistent calcium (Cav1.3) and sodium currents spontaneously develop in motoneurons of chronic spinal rats and these enable large, rapidly activated plateaus that ultimately lead to spasticity. PMID- 12724370 TI - Genetic relatedness and phenotypic characteristics of Treponema associated with human periodontal tissues and ruminant foot disease. AB - Treponema have been implicated recently in the pathogenesis of digital dermatitis (DD) and contagious ovine digital dermatitis (CODD) that are infectious diseases of bovine and ovine foot tissues, respectively. Previous analyses of treponemal 16S rDNA sequences, PCR-amplified directly from DD or CODD lesions, have suggested relatedness of animal Treponema to some human oral Treponema species isolated from periodontal tissues. In this study a range of adhesion and virulence-related properties of three animal Treponema isolates have been compared with representative human oral strains of Treponema denticola and Treponema vincentii. In adhesion assays using biotinylated treponemal cells, T. denticola cells bound in consistently higher numbers to fibronectin, laminin, collagen type I, gelatin, keratin and lactoferrin than did T. vincentii or animal Treponema isolates. However, animal DD strains adhered to fibrinogen at equivalent or greater levels than T. denticola. All Treponema strains bound to the amino-terminal heparin I/fibrin I domain of fibronectin. 16S rDNA sequence analyses placed ovine strain UB1090 and bovine strain UB1467 within a cluster that was phylogenetically related to T. vincentii, while ovine strain UB1466 appeared more closely related to T. denticola. These observations correlated with phenotypic properties. Thus, T. denticola ATCC 35405, GM-1, and Treponema UB1466 had similar outer-membrane protein profiles, produced chymotrypsin-like protease (CTLP), trypsin-like protease and high levels of proline iminopeptidase, and co aggregated with human oral bacteria Porphyromonas gingivalis and Streptococcus crista. Conversely, T. vincentii ATCC 35580, D2A-2, and animal strains UB1090 and UB1467 did not express CTLP or trypsin-like protease and did not co-aggregate with P. gingivalis or S. crista. Taken collectively, these results suggest that human oral-related Treponema have broad host specificity and that similar control or preventive strategies might be developed for human and animal Treponema associated infections. PMID- 12724371 TI - Fluorescence in situ hybridization (FISH) for direct visualization of bacteria in periapical lesions of asymptomatic root-filled teeth. AB - Whether micro-organisms can live in periapical endodontic lesions of asymptomatic teeth is under debate. The aim of the present study was to visualize and identify micro-organisms within periapical lesions directly, using fluorescence in situ hybridization (FISH) in combination with epifluorescence and confocal laser scanning microscopy (CLSM). Thirty-nine periapical lesions were surgically removed, fixed, embedded in cold polymerizing resin and sectioned. The probe EUB 338, specific for the domain Bacteria, was used together with a number of species specific 16S rRNA-directed oligonucleotide probes to identify bacteria. To control non-specific binding of EUB 338, probe NON 338 was used. Alternatively, DAPI (4',6'-diamidino-2-phenylindole) staining was applied to record prokaryotic and eukaryotic DNA in the specimens. Hybridization with NON 338 gave no signals despite background fluorescence of the tissue. The eubacterial probe showed bacteria of different morphotypes in 50 % of the lesions. Rods, spirochaetes and cocci were spread out in areas of the tissue while other parts seemed bacteria free. Bacteria were also seen to co-aggregate inside the tissue, forming microcolonies. Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis and treponemes of phylogenetic Group I were detected with specific probes. In addition, colonies with Streptococcus spp. were seen in some lesions. A number of morphotypes occurred that could not be identified with the specific probes used, indicating the presence of additional bacterial species. CLSM confirmed that bacteria were located in different layers of the tissue. Accordingly, the FISH technique demonstrated mixed consortia of bacteria consisting of rods, spirochaetes and cocci in asymptomatic periapical lesions of root-filled teeth. PMID- 12724372 TI - SseA is a chaperone for the SseB and SseD translocon components of the Salmonella pathogenicity-island-2-encoded type III secretion system. AB - The type III secretion system (TTSS) encoded by the Salmonella pathogenicity island 2 (SPI-2) is required for bacterial replication inside macrophages and for systemic infection in mice. Many TTSS secreted proteins, including effectors and components of the translocon, require chaperones which promote their stability, prevent their premature interactions or facilitate their secretion. In this study, the function of the first gene (sseA) of one of the SPI-2 operons (sseA-G) was investigated. This operon includes genes that encode translocon components (SseB, SseC and SseD), translocated proteins (SseF and SseG) and putative chaperones (SscA and SscB). sseA encodes a 12.5 kDa protein with a C-terminal region with the potential to form a coiled-coil structure, but no sequence similarity to other proteins. Mutation of sseA results in severe virulence attenuation and an intracellular replication defect. It is shown here that SseA is not a secreted protein, but is required for SPI-2-dependent translocation of two effector proteins (SifA and PipB). Furthermore, the translocon components SseB and SseD were not detected in an sseA mutant strain. By using a yeast two hybrid assay and column binding experiments, it is demonstrated that SseA interacts directly with SseB and SseD. These results indicate that SseA is a chaperone for SseB and SseD. The inability of an sseA mutant to assemble the SPI 2 TTSS translocon accounts for its high level of virulence attenuation in vivo. To the authors' knowledge, this is the first chaperone described for the SPI-2 TTSS. PMID- 12724373 TI - Immunological and genetic characterization of Borrelia burgdorferi BapA and EppA proteins. AB - A large majority of examined Lyme disease spirochaete isolates were demonstrated to contain one or both of the paralogous genes bapA and eppA. Immunological analyses of serum samples collected from infected patients coupled with comparative sequence analyses indicated that bapA gene sequences are quite stable but the encoded proteins do not provoke a strong immune response in most individuals. Conversely, EppA proteins are much more antigenic but vary widely in sequence between different bacteria. Considerable evidence of insertion, deletion and other mutations within eppA genes was observed. A number of significant recombination events were also found to have occurred in regions flanking bapA genes, while the genes themselves rarely exhibited evidence of mutation, suggesting strong selective pressure to maintain BapA sequences within narrow limits. Data from these and other studies suggest important roles for BapA and EppA during the Borrelia burgdorferi infectious cycle. PMID- 12724374 TI - Alginate gene expression by Pseudomonas syringae pv. tomato DC3000 in host and non-host plants. AB - Pseudomonas syringae produces the exopolysaccharide alginate, a copolymer of mannuronic and guluronic acid. Although alginate has been isolated from plants infected by P. syringae, the signals and timing of alginate gene expression in planta have not been described. In this study, an algD : : uidA transcriptional fusion, designated pDCalgDP, was constructed and used to monitor alginate gene expression in host and non-host plants inoculated with P. syringae pv. tomato DC3000. When leaves of susceptible collard plants were spray-inoculated with DC3000(pDCalgDP), algD was activated within 72 h post-inoculation (p.i.) and was associated with the development of water-soaked lesions. In leaves of the susceptible tomato cv. Rio Grande-PtoS, algD activity was lower than in collard and was not associated with water-soaking. The expression of algD was also monitored in leaves of tomato cv. Rio Grande-PtoR, which is resistant to P. syringae pv. tomato DC3000. Within 12 h p.i., a microscopic hypersensitive response (micro-HR) was observed in Rio Grande-PtoR leaves spray-inoculated with P. syringae pv. tomato DC3000(pDCalgDP). As the HR progressed, histochemical staining indicated that individual bacterial cells on the surface of resistant tomato leaves were expressing algD. These results indicate that algD is expressed in both susceptible (e.g. collard, tomato) and resistant (Rio Grande-PtoR) host plants. The expression of algD in an incompatible host-pathogen interaction was further explored by monitoring transcriptional activity in leaves of tobacco, which is not a host for P. syringae pv. tomato. In tobacco inoculated with DC3000(pDCalgDP), an HR was evident within 12 h p.i., and algD expression was evident within 8-12 h p.i. However, when tobacco was inoculated with an hrcC mutant of DC3000, the HR did not occur and algD expression was substantially lower. These results suggest that signals that precede the HR may stimulate alginate gene expression in P. syringae. Histochemical staining with nitro blue tetrazolium indicated that the superoxide anion () is a signal for algD activation in planta. This study indicates that algD is expressed when P. syringae attempts to colonize both susceptible and resistant plant hosts. PMID- 12724375 TI - Diurnal expression of hetR and diazocyte development in the filamentous non heterocystous cyanobacterium Trichodesmium erythraeum. AB - The marine non-heterocystous cyanobacterium Trichodesmium fixes atmospheric N(2) aerobically in light. In situ immunolocalization/light microscopy of NifH revealed that lighter, non-granulated cell regions observed correspond to the nitrogenase-containing diazocyte clusters in Trichodesmium IMS101. The number of diazocyte clusters per trichome varied from 0 to 4 depending on trichome length. The constant percentage of diazocytes (approx. 15 %) in cultured strains and five natural populations suggests a developmentally regulated differentiation process. Real-time RT-PCR showed that ntcA, encoding the global nitrogen regulator in cyanobacteria, and hetR, the key regulatory gene in heterocyst differentiation, are both constitutively expressed during a 12 h/12 h light/dark cycle. hetR in addition showed a distinct peak in the dark (close to midnight) while nifH expression commenced 6-8 h later. The expression of all three genes was negatively affected by addition of ammonia. Some early heterocyst differentiation genes were also identified in the genome of Trichodesmium. The data suggest that hetR and ntcA may be required for development and function of diazocytes in Trichodesmium. PMID- 12724376 TI - Calcium release from Synechocystis cells induced by depolarization of the plasma membrane: MscL as an outward Ca2+ channel. AB - Cells of the cyanobacterium Synechocystis sp. PCC 6803 are equipped with a mechanosensitive ion channel MscL that is located in their plasma membrane. However, the exact function of the channel in this freshwater cyanobacterium is unknown. This study shows that cells of Synechocystis are capable of releasing Ca(2+) in response to depolarization of the plasma membrane by the K(+) ionophore valinomycin in the presence of K(+) or by tetraphenylphosphonium (TPP(+)). A fluorescent dye, diS-C(3)-(5), sensitive to membrane potential and the metallochromic Ca(2+) indicator arsenazo III were used to follow the plasma membrane depolarization and the Ca(2+) release, respectively. The Ca(2+) release from wild-type cells was temperature-dependent and it was strongly inhibited by the Ca(2+) channel blocker verapamil and by the mechanosensitive channel blocker amiloride. In MscL-deficient cells, Ca(2+) release was about 50 % of that from the wild-type cells. The mutant cells had lost temperature sensitivity of Ca(2+) release completely. However, verapamil and amiloride inhibited Ca(2+) release from these cells in same manner as in the wild-type cells. This suggests the existence of additional Ca(2+) transporters in Synechocystis, probably of a mechanosensitive nature. Evidence for the putative presence of intracellular Ca(2+) stores in the cells was obtained by following the increase in fluorescence intensity of the Ca(2+) indicator chlortetracycline. These results suggest that the MscL of Synechocystis might operate as a verapamil/amiloride-sensitive outward Ca(2+) channel that is involved in the plasma-membrane depolarization induced Ca(2+) release from the cells under temperature stress conditions. PMID- 12724377 TI - A computer investigation of chemically mediated detachment in bacterial biofilms. AB - A three-dimensional computer model was used to evaluate the effect of chemically mediated detachment on biofilm development in a negligible-shear environment. The model, BacLAB, combines conventional diffusion-reaction equations for chemicals with a cellular automata algorithm to simulate bacterial growth, movement and detachment. BacLAB simulates the life cycle of a bacterial biofilm from its initial colonization of a surface to the development of a mature biofilm with cell areal densities comparable to those in the laboratory. A base model founded on well established transport equations that are easily adaptable to investigate conjectures at the biological level has been created. In this study, the conjecture of a detachment mechanism involving a bacterially produced chemical detachment factor in which high local concentrations of this detachment factor cause the bacteria to detach from the biofilm was examined. The results show that the often observed 'mushroom'-shaped structure can occur if detachment events create voids so that the remaining attached cells look like mushrooms. PMID- 12724379 TI - Heterologous expression of laccase cDNA from Ceriporiopsis subvermispora yields copper-activated apoprotein and complex isoform patterns. AB - Analysis of genomic clones encoding a putative laccase in homokaryon strains of Ceriporiopsis subvermispora led to the identification of an allelic variant of the previously described lcs-1 gene. A cDNA clone corresponding to this gene was expressed in Aspergillus nidulans and in Aspergillus niger. Enzyme assays and Western blots showed that both hosts secreted active laccase. Relative to the isozymic forms of the native C. subvermispora enzyme, the A. niger-produced laccase had a higher molecular mass and gave a single band on IEF gels. In contrast, A. nidulans transformants secreted several isoforms remarkably similar to those of the native system. Considered together with previously reported Southern blots and protein sequencing, expression in A. nidulans supports the view that C. subvermispora has a single laccase gene and that multiple isoforms result from post-translational processes. In addition, several lines of evidence strongly suggest that under copper limitation, A. nidulans secretes apoprotein which can be reconstituted by a short incubation with Cu(I) and to a lesser extent with Cu(II). PMID- 12724378 TI - Only one catalase, katG, is detectable in Rhizobium etli, and is encoded along with the regulator OxyR on a plasmid replicon. AB - The plasmid-borne Rhizobium etli katG gene encodes a dual-function catalase peroxidase (KatG) (EC 1.11.1.7) that is inducible and heat-labile. In contrast to other rhizobia, katG was shown to be solely responsible for catalase and peroxidase activity in R. etli. An R. etli mutant that did not express catalase activity exhibited increased sensitivity to hydrogen peroxide (H(2)O(2)). Pre exposure to a sublethal concentration of H(2)O(2) allowed R. etli to adapt and survive subsequent exposure to higher concentrations of H(2)O(2). Based on a multiple sequence alignment with other catalase-peroxidases, it was found that the catalytic domains of the R. etli KatG protein had three large insertions, two of which were typical of KatG proteins. Like the katG gene of Escherichia coli, the R. etli katG gene was induced by H(2)O(2) and was important in sustaining the exponential growth rate. In R. etli, KatG catalase-peroxidase activity is induced eightfold in minimal medium during stationary phase. It was shown that KatG catalase-peroxidase is not essential for nodulation and nitrogen fixation in symbiosis with Phaseolus vulgaris, although bacteroid proteome analysis indicated an alternative compensatory mechanism for the oxidative protection of R. etli in symbiosis. Next to, and divergently transcribed from the catalase promoter, an ORF encoding the regulator OxyR was found; this is the first plasmid-encoded oxyR gene described so far. Additionally, the katG promoter region contained sequence motifs characteristic of OxyR binding sites, suggesting a possible regulatory mechanism for katG expression. PMID- 12724380 TI - Isolation and characterization of two specific regulatory Aspergillus niger mutants shows antagonistic regulation of arabinan and xylan metabolism. AB - This paper describes two Aspergillus niger mutants (araA and araB) specifically disturbed in the regulation of the arabinanase system in response to the presence of L-arabinose. Expression of the three known L-arabinose-induced arabinanolytic genes, abfA, abfB and abnA, was substantially decreased or absent in the araA and araB strains compared to the wild-type when incubated in the presence of L arabinose or L-arabitol. In addition, the intracellular activities of L-arabitol dehydrogenase and L-arabinose reductase, involved in L-arabinose catabolism, were decreased in the araA and araB strains. Finally, the data show that the gene encoding D-xylulose kinase, xkiA, is also under control of the arabinanolytic regulatory system. L-Arabitol, most likely the true inducer of the arabinanolytic and L-arabinose catabolic genes, accumulated to a high intracellular concentration in the araA and araB mutants. This indicates that the decrease of expression of the arabinanolytic genes was not due to lack of inducer accumulation. Therefore, it is proposed that the araA and araB mutations are localized in positive-acting components of the regulatory system involved in the expression of the arabinanase-encoding genes and the genes encoding the L arabinose catabolic pathway. PMID- 12724381 TI - Response to high osmotic conditions and elevated temperature in Saccharomyces cerevisiae is controlled by intracellular glycerol and involves coordinate activity of MAP kinase pathways. AB - In the yeast Saccharomyces cerevisiae, response to an increase in external osmolarity is mediated by the HOG (high osmolarity glycerol) MAP kinase pathway. HOG pathway mutant strains display osmosensitive phenotypes. Recently evidence has been obtained that the osmosensitivity of HOG pathway mutants is reduced during growth at elevated temperature (37 degrees C). A notable exception is the ste11ssk2ssk22 mutant, which displays hypersensitivity to osmotic stress at 37 degrees C. This paper reports that overexpression of FPS1 or GPD1 (encoding the glycerol transport facilitator and glycerol-3-phosphate dehydrogenase, respectively, and both affecting intracellular glycerol levels) reduces the hypersensitivity to osmotic stress of ste11ssk2ssk22 at 37 degrees C. Although in this particular HOG pathway mutant a correlation between suppression of the phenotype and glycerol content could be demonstrated, the absolute level of intracellular glycerol per se does not determine whether a strain is osmosensitive or not. Rather, evidence was obtained that the glycerol level may have an indirect effect, viz. by influencing signalling through the PKC (protein kinase C) MAP kinase pathway, which plays an important role in maintenance of cellular integrity. In order to validate the data obtained with a HOG pathway mutant strain for wild-type yeast cells, MAP kinase signalling under different growth conditions was examined in wild-type strains. PKC pathway signalling, which is manifest at elevated growth temperature by phosphorylation of MAP kinase Mpk1p, is rapidly lost when cells are shifted to high external osmolarity conditions. Expression of bck1-20 or overexpression of WSC3 in wild-type cells resulted in restoration of PKC signalling. Both PKC and HOG signalling, cell wall phenotypes and high osmotic stress responses in wild-type cells were found to be influenced by the growth temperature. The data taken together indicate the intricate interdependence of growth temperature, intracellular glycerol, cell wall structure and MAP kinase signalling in the hyperosmotic stress response of yeast. PMID- 12724382 TI - Synergistic inhibition of APC/C by glucose and activated Ras proteins can be mediated by each of the Tpk1-3 proteins in Saccharomyces cerevisiae. AB - Proteolysis triggered by the anaphase-promoting complex/cyclosome (APC/C) is essential for the progression through mitosis. APC/C is a highly conserved ubiquitin ligase whose activity is regulated during the cell cycle by various factors, including spindle checkpoint components and protein kinases. The cAMP dependent protein kinase (PKA) was identified as negative regulator of APC/C in yeast and mammalian cells. In the yeast Saccharomyces cerevisiae, PKA activity is induced upon glucose addition or by activated Ras proteins. This study shows that glucose and the activated Ras2(Val19) protein synergistically inhibit APC/C function via the cAMP/PKA pathway in yeast. Remarkably, Ras2 proteins defective in the interaction with adenylate cyclase fail to influence APC/C, implying that its function is regulated exclusively by PKA, but not by alternative Ras pathways. Furthermore, it is shown that the three PKAs in yeast, Tpk1, Tpk2 and Tpk3, have redundant functions in regulating APC/C in response to glucose medium. Single or double deletions of TPK genes did not prevent inhibition of APC/C, suggesting that each of the Tpk proteins can take over this function. However, Tpk2 seems to inhibit APC/C function more efficiently than Tpk1 and Tpk3. Finally, evidence is provided that Cdc20 is involved in APC/C regulation by the cAMP/PKA pathway. PMID- 12724383 TI - An iron-regulated gene required for utilization of aerobactin as an exogenous siderophore in Vibrio parahaemolyticus. AB - A previous investigation using the Fur titration assay system showed that Vibrio parahaemolyticus possesses a gene encoding a protein homologous to IutA, the outer-membrane receptor for ferric aerobactin in Escherichia coli. In this study, a 5.6 kb DNA region from the V. parahaemolyticus WP1 genome was cloned and two entire genes, iutA and alcD homologues, were identified which are absent from Vibrio cholerae genomic sequences. The V. parahaemolyticus IutA and AlcD proteins share 43 % identity with the Escherichia coli IutA protein and 24 % identity with the Bordetella bronchiseptica AlcD protein of unknown function, respectively. Primer extension analysis revealed that the iutA gene is transcribed in response to low-iron availability from a putative promoter overlapped with a sequence resembling a consensus E. coli Fur-binding sequence. In agreement with the above finding, V. parahaemolyticus effectively utilized exogenously supplied aerobactin for growth under iron-limiting conditions. Moreover, insertional inactivation of iutA impaired growth in the presence of aerobactin and incapacitated the outer membrane fraction from iron-deficient cells for binding (55)Fe-labelled aerobactin. These results indicate that the V. parahaemolyticus iutA homologue encodes an outer-membrane protein which functions as the receptor for ferric aerobactin. Southern blot analysis revealed that the iutA homologues are widely distributed in clinical and environmental isolates of V. parahaemolyticus. However, additional genes required for ferric aerobactin transport across the inner membrane remain to be clarified. PMID- 12724384 TI - Genetic organization of an Acinetobacter baumannii chromosomal region harbouring genes related to siderophore biosynthesis and transport. AB - The Acinetobacter baumannii 8399 clinical isolate secretes dihydroxybenzoic acid (DHBA) and a high-affinity catechol siderophore, which is different from other bacterial iron chelators already characterized. Complementation assays with enterobactin-deficient Escherichia coli strains led to the isolation of a cosmid clone containing A. baumannii 8399 genes required for the biosynthesis and activation of DHBA. Accordingly, the cloned fragment harbours a dhbACEB polycistronic operon encoding predicted proteins highly similar to several bacterial proteins required for DHBA biosynthesis from chorismic acid. Genes encoding deduced proteins related to the E. coli Fes and the Bacillus subtilis DhbF proteins, and a putative Yersinia pestis phosphopantetheinyl transferase, all of them involved in the assembly and utilization of catechol siderophores in other bacteria, were found next to the dhbACEB locus. This A. baumannii 8399 gene cluster also contained the om73, p45 and p114 predicted genes encoding proteins potentially involved in transport of ferric siderophore complexes. The deduced products of the p114 and p45 genes are putative membrane proteins that belong to the RND and MFS efflux pump proteins, respectively. Interestingly, P45 is highly related to the E. coli P43 (EntS) protein that participates in the secretion of enterobactin. Although P114 is similar to other bacterial efflux pump proteins involved in antibiotic resistance, its genetic arrangement within this A. baumannii 8399 locus is different from that described in other bacteria. The product of om73 is a Fur- and iron-regulated surface-exposed outer-membrane protein. These characteristics together with the presence of a predicted TonB box and its high similarity to other siderophore receptors indicate that OM73 plays such a role in A. baumannii 8399. The 184 nt om73-p114 intergenic region contains promoter elements that could drive the expression of these divergently transcribed genes, all of which are in close proximity to almost perfect Fur boxes. This arrangement explains the iron- and Fur-regulated expression of om73, and provides strong evidence for a similar regulation for the expression of p114. PMID- 12724385 TI - Limitations of the widely used GAM42a and BET42a probes targeting bacteria in the Gammaproteobacteria radiation. AB - The 23S rRNA-targeted probes GAM42a and BET42a provided equivocal results with the uncultured gammaproteobacterium 'Candidatus Competibacter phosphatis' where some cells bound GAM42a and other cells bound BET42a in fluorescence in situ hybridization (FISH) experiments. Probes GAM42a and BET42a span positions 1027 1043 in the 23S rRNA and differ from each other by one nucleotide at position 1033. Clone libraries were prepared from PCR products spanning the 16S rRNA genes, intergenic spacer region and 23S rRNA genes from two mixed cultures enriched in 'Candidatus C. phosphatis'. With individual clone inserts, the 16S rDNA portion was used to confirm the source organism as 'Candidatus C. phosphatis' and the 23S rDNA portion was used to determine the sequence of the GAM42a/BET42a probe target region. Of the 19 clones sequenced, 8 had the GAM42a probe target (T at position 1033) and 11 had G at position 1033, the only mismatch with GAM42a. However, none of the clones had the BET42a probe target (A at 1033). Non-canonical base-pairing between the 23S rRNA of 'Candidatus C. phosphatis' with G at position 1033 and GAM42a (G-A) or BET42a (G-T) is likely to explain the probing anomalies. A probe (GAM42_C1033) was optimized for use in FISH, targeting cells with G at position 1033, and was found to highlight not only some 'Candidatus C. phosphatis' cells, but also other bacteria. This demonstrates that there are bacteria in addition to 'Candidatus C. phosphatis' with the GAM42_C1033 probe target and not the BET42a or GAM42a probe target. PMID- 12724386 TI - Modification of the signal sequence cleavage site of listeriolysin O does not affect protein secretion but impairs the virulence of Listeria monocytogenes. AB - Listeriolysin O (LLO, hly-encoded), a major virulence factor secreted by the bacterial pathogen Listeria monocytogenes, is synthesized as a precursor of 529 residues. To impair LLO secretion, the four residues of the predicted signal sequence cleavage site (EA-KD) were deleted and the mutant LLO protein was expressed in a hly-negative derivative of L. monocytogenes. Unexpectedly, the mutant protein was secreted in normal amounts in the culture supernatant and was fully haemolytic. N-terminal sequencing of the secreted LLO molecule revealed that N-terminal processing of the preprotein occurred three residues downstream of the natural cleavage site. L. monocytogenes expressing this truncated LLO showed a reduced capacity to disrupt the phagosomal membranes of bone marrow macrophages and of hepatocytes; and the mutant strain showed a 100-fold decrease in virulence in the mouse model. These results suggest that the first N-terminal residues of mature LLO participate directly in phagosomal escape and bacterial infection. PMID- 12724387 TI - Adhesins encoded by the gingipain genes of Porphyromonas gingivalis are responsible for co-aggregation with Prevotella intermedia. AB - Co-aggregation among bacterial cells caused by the adherence of one bacterial species to another is a potential colonization mechanism. Several putative aggregation factors for co-aggregation between Porphyromonas (Por.) gingivalis and Prevotella (Pre.) intermedia were partially purified from Por. gingivalis vesicles by gel filtration and affinity chromatography. Antisera against the aggregation factors were made. Analysis using these antisera revealed that 18 and 44 kDa proteins might be responsible for Por. gingivalis vesicle-mediated aggregation of Pre. intermedia. Using antiserum against the 18 kDa protein, the DNA region encoding it was cloned from Por. gingivalis genomic DNA. Sequence analysis revealed that the DNA region was located within the rgpA and kgp genes, encoding Arg-gingipain (Rgp) and Lys-gingipain (Kgp), respectively, and it encoded non-catalytic adhesin domain regions, namely a C-terminal portion of HGP15, the entire HGP17 sequence and an N-terminal portion of HGP27. A portion of the DNA sequence was also found in the haemagglutinin A (hagA) gene. A recombinant glutathione S-transferase (GST)-HGP17 fusion protein reacted to antiserum against the 18 kDa protein and Pre. intermedia cells could adhere to GST-HGP17-conjugated Sepharose 4B beads, indicating that the HGP17 domain protein is responsible for Por. gingivalis vesicle-mediated aggregation of Pre. intermedia. PMID- 12724388 TI - n-Hexane sensitivity of Escherichia coli due to low expression of imp/ostA encoding an 87 kDa minor protein associated with the outer membrane. AB - Most Escherichia coli strains are resistant to n-hexane. E. coli OST4251 is a n hexane-sensitive strain that was constructed by transferring the n-hexane sensitive phenotype from a n-hexane-sensitive strain by P1 transduction. OST4251 is resistant to diphenyl ether, which is less harmful than n-hexane to micro organisms. The genetic determinant responsible for this subtle difference in the solvent resistance is mapped at 1.2 min on the E. coli chromosome. Nucleotide sequence analysis showed that IS2 and IS5 had integrated upstream of the imp/ostA structural gene in OST4251. The integration of IS2 decreased the activity of the imp/ostA promoter. A product of the gene was identified immunologically as an 87 kDa minor protein associated with the outer membrane. Upon transformation with plasmids containing the imp/ostA gene, OST4251 produced a high level of the gene product in the membrane and acquired n-hexane resistance. Thus, the low level of promoter activity resulted in low Imp production and the n-hexane-sensitivity phenotype. It is likely that the gene product contributes to n-hexane resistance by reducing the influx of n-hexane. PMID- 12724390 TI - An inducible tellurite-resistance operon in Proteus mirabilis. AB - Tellurite resistance (Te(r)) is widespread in nature and it is shown here that the natural resistance of Proteus mirabilis to tellurite is due to a chromosomally located orthologue of plasmid-borne ter genes found in enteric bacteria. The P. mirabilis ter locus (terZABCDE) was identified in a screen of Tn5lacZ-generated mutants of which one contained an insertion in terC. The P. mirabilis terC mutant displayed increased susceptibility to tellurite (Te(s)) and complementation with terC carried on a multicopy plasmid restored high-level Te(r). Primer extension analysis revealed a single transcriptional start site upstream of terZ, but only with RNA harvested from bacteria grown in the presence of tellurite. Northern blotting and reverse transcriptase-PCR (RT-PCR) analyses confirmed that the ter operon was inducible by tellurite and to a lesser extent by oxidative stress inducers such as hydrogen peroxide and methyl viologen (paraquat). Direct and inverted repeat sequences were identified in the ter promoter region as well as motifs upstream of the -35 hexamer that resembled OxyR binding sequences. Finally, the 390 bp intergenic promoter region located between orf3 and terZ showed no DNA sequence identity with any other published ter sequences, whereas terZABCDE genes exhibited 73-85 % DNA sequence identity. The ter operon was present in all clinical isolates of P. mirabilis and Proteus vulgaris tested and is inferred for Morganella and Providencia spp. based on screening for high level Te(r) and preliminary PCR analysis. Thus, a chromosomally located inducible tellurite resistance operon appears to be a common feature of the genus Proteus. PMID- 12724389 TI - Regulation of expression of the cyanide-insensitive terminal oxidase in Pseudomonas aeruginosa. AB - The regulation of the cyanide-insensitive oxidase (CIO) in Pseudomonas aeruginosa, a bacterium that can synthesize HCN, is reported. The expression of a cioA-lacZ transcriptional fusion, CioA protein levels and CIO activity were low in exponential phase but induced about fivefold upon entry into stationary phase. Varying the O(2) transfer coefficient from 11.5 h(-1) to 87.4 h(-1) had no effect on CIO expression and no correlation was observed between CIO induction and the dissolved O(2) levels in the growth medium. However, a mutant deleted for the O(2)-sensitive transcriptional regulator ANR derepressed CIO expression in an O(2)-sensitive manner, with the highest induction occurring under low-O(2) conditions. Therefore, CIO expression can respond to a signal generated by low O(2) levels, but this response is normally kept in check by ANR repression. ANR may play an important role in preventing overexpression of the CIO in relation to other terminal oxidases. A component present in spent culture medium was able to induce CIO expression. However, experiments with purified N-butanoyl-L-homoserine lactone or N-(3-oxododecanoyl)homoserine lactone ruled out a role for these quorum-sensing molecules in the control of CIO expression. Cyanide was a potent inducer of the CIO at physiologically relevant concentrations and experiments using spent culture medium from a DeltahcnB mutant, which is unable to synthesize cyanide, showed that cyanide was the inducing factor present in P. aeruginosa spent culture medium. However, the finding that in a DeltahcnB mutant cioA-lacZ expression was induced normally upon entry into stationary phase indicated that cyanide was not the endogenous inducer of the terminal oxidase. The authors suggest that the failure of O(2) to have an effect on CIO expression in the wild type can be explained either by the requirement for an additional, stationary phase-specific inducing signal or by the loss of an exponential-phase-specific repressing signal. PMID- 12724392 TI - Proteome analysis of extracellular proteins regulated by the las and rhl quorum sensing systems in Pseudomonas aeruginosa PAO1. AB - The las and rhl quorum sensing (QS) systems regulate the expression of several genes in response to cell density changes in Pseudomonas aeruginosa. Many of these genes encode surface-associated or secreted virulence factors. Proteins from stationary phase culture supernatants were collected from wild-type and P. aeruginosa PAO1 mutants deficient in one or more of the lasRI, rhlRI and vfr genes and analysed using two-dimensional gel electrophoresis. All mutants released significantly lower amounts of protein than the wild-type. Protein spot patterns from each strain were compared using image analysis and visible spot differences were identified using mass spectrometry. Several previously unknown QS-regulated proteins were characterized, including an aminopeptidase (PA2939), an endoproteinase (PrpL) and a unique 'hypothetical' protein (PA0572), which could not be detected in the culture supernatants of Deltalas mutants, although they were unaffected in Deltarhl mutants. Chitin-binding protein (CbpD) and a hypothetical protein (PA4944) with similarity to host factor I (HF-I) could not be detected when any of the lasRI or rhlRI genes were disrupted. Fourteen proteins were present at significantly greater levels in the culture supernatants of QS mutants, suggesting that QS may also negatively control the expression of some genes. Increased levels of two-partner secretion exoproteins (PA0041 and PA4625) were observed and may be linked to increased stability of their cognate transporters in a QS-defective background. Known QS-regulated extracellular proteins, including elastase (lasB), LasA protease (lasA) and alkaline metalloproteinase (aprA) were also detected. PMID- 12724391 TI - Detection and analysis of transpositionally active head-to-tail dimers in three additional Escherichia coli IS elements. AB - This study demonstrates that Escherichia coli insertion elements IS3, IS150 and IS186 are able to form transpositionally active head-to-tail dimers which show similar structure and transpositional activity to the dimers of IS2, IS21 and IS30. These structures arise by joining of the left and right inverted repeats (IRs) of two elements. The resulting junction includes a spacer region (SR) of a few base pairs derived from the flanking sequence of one of the reacting IRs. Head-to-tail dimers of IS3, IS150 and IS186 are unstable due to their transpositional activity. They can be resolved in two ways that seem to form a general rule for those elements reported to form dimers. One way is a site specific process (dimer dissolution) which is accompanied by the loss of one IS copy along with the SR. The other is 'classical' transposition where the joined ends integrate into the target DNA. In intramolecular transposition this often gives rise to deletion formation, whereas in intermolecular transposition it gives rise to replicon fusion. The results presented for IS3, IS150 and IS186 are in accordance with the IS dimer model, which is in turn consistent with models based on covalently closed minicircles. PMID- 12724393 TI - Molecular analysis of mutS expression and mutation in natural isolates of pathogenic Escherichia coli. AB - Deficiencies in the MutS protein disrupt methyl-directed mismatch repair (MMR), generating a mutator phenotype typified by high mutation rates and promiscuous recombination. How such deficiencies might arise in the natural environment was determined by analysing pathogenic strains of Escherichia coli. Quantitative Western immunoblotting showed that the amount of MutS in a wild-type strain of the enterohaemorrhagic pathogen E. coli O157 : H7 decreased about 26-fold in stationary-phase cells as compared with the amount present during exponential phase growth. The depletion of MutS in O157 : H7 is significantly greater than that observed for a laboratory-attenuated E. coli K-12 strain. In the case of stable mutators, mutS defects in strains identified among natural isolates were analysed, including two E. coli O157 : H7 strains, a diarrhoeagenic E. coli O55 : H7 strain, and a uropathogenic strain from the E. coli reference (ECOR) collection. No MutS could be detected in the four strains by Western immunoblot analyses. RNase T2 protection assays showed that the strains were either deficient in mutS transcripts or produced transcripts truncated at the 3' end. Nucleotide sequence analysis revealed extensive deletions in the mutS region of three strains, ranging from 7.5 to 17.3 kb relative to E. coli K-12 sequence, while the ECOR mutator contained a premature stop codon in addition to other nucleotide changes in the mutS coding sequence. These results provide insights into the status of the mutS gene and its product in pathogenic strains of E. coli. PMID- 12724394 TI - The LicT protein acts as both a positive and a negative regulator of loci within the bgl regulon of Streptococcus mutans. AB - An open reading frame (ORF) that would encode a putative antiterminator protein (LicT) of the BglG family was identified in the genomic DNA sequence of Streptococcus mutans. A DNA sequence that would encode a potential ribonucleic antiterminator (RAT) site in the mRNA at which the putative antitermination protein LicT would bind was located immediately downstream from this ORF. These putative antitermination components are upstream of a glucose-independent beta glucoside-utilization system that is responsible for aesculin utilization by S. mutans NG8 in the presence of glucose. It was hypothesized that these putative regulatory components were an important mechanism that was involved with the controlled expression of the S. mutans bglP locus. A strain of S. mutans containing a licT : : Omega-Kan2 insertional mutation was created. This strain could not hydrolyse aesculin in the presence of glucose. The transcriptional activity associated with other genes from the bgl regulon was determined in the licT : : Omega-Kan2 genetic background using lacZ transcriptional fusions and beta-galactosidase assays to determine the effect of LicT on these loci. The LicT protein had no significant effect on the expression of the bglC promoter, a regulator of the bglA locus. However, it is essential for the optimal expression of bglP. These data correlate with the phenotype observed on aesculin plates for the S. mutans wild-type strain NG8 and the licT : : Omega-Kan2 strain. Thus, the glucose-independent beta-glucoside-specific phosphotransferase system (PTS) regulon in S. mutans relies on LicT for BglP expression and, in turn, aesculin transport in the presence of glucose. Interestingly, LicT also seems to negatively regulate the expression of the bglA promoter region. In addition, the presence of the S. mutans licT gene has been shown to be able to activate a cryptic beta-glucoside-specific operon found in Escherichia coli. PMID- 12724395 TI - Inactivation of mshB, a key gene in the mycothiol biosynthesis pathway in Mycobacterium smegmatis. AB - The mshB gene encoding N-acetyl-1-D-myo-inosityl-2-amino-2-deoxy-alpha-D glucopyranoside deacetylase (MshB) is a key enzyme in mycothiol biosynthesis. Disruption of mshB in Mycobacterium smegmatis resulted in decreased production of mycothiol (5-10 % of the parent strain mc(2)155) but did not abolish mycothiol synthesis completely. Complementation of the MshB(-) mutants with the mshB gene resulted in increased mycothiol production towards the exponential and stationary phases of the bacterial growth cycle. These results suggest that another enzyme is capable of mycothiol biosynthesis by providing N-acetylglucosaminylinositol deacetylation activity in the absence of MshB. One of the candidate enzymes capable of carrying out such reactions is the MshB orthologue mycothiol amide hydrolase, MCA. However, epichromosomal expression of mca in the MshB(-) mutants did not restore mycothiol levels to the level of the parent strain. Unlike other mutants, which have little or no detectable levels of mycothiol, the MshB(-) mutant did not exhibit increased resistance to isoniazid. However, the MshB(-) mutant was resistant to ethionamide. Phenotypic analysis of other mutants lacking mycothiol revealed that MshA(-) mutants also exhibit ethionamide resistance but that a MshC(-)mutant was sensitive to ethionamide, suggesting that mycothiol or its early intermediates influence ethionamide activation. PMID- 12724396 TI - Limited regions of homology between linear and circular plasmids encoding methylenomycin biosynthesis in two independently isolated streptomycetes. AB - pSV1 is a plasmid in Streptomyces violaceoruber SANK95570 that carries the methylenomycin biosynthetic (mmy) gene cluster. An ordered cosmid map and an EcoRI map have been constructed for pSV1, confirming that pSV1 is a 163 kb circular plasmid. The mmy gene cluster has been found on three different replicon structures; the circular plasmid pSV1, the 356 kb linear plasmid SCP1 and, via SCP1 integration, the linear chromosome of Streptomyces coelicolor A3(2). Comparison of pSV1 and SCP1 sequences revealed that the two plasmids have homology to each other only around the mmy and parAB regions, eliminating models in which pSV1 was generated by circularization of SCP1 or vice versa. It is likely that the mmy gene cluster was horizontally transferred as a set together with the parAB region in the comparatively recent evolutionary past. PMID- 12724397 TI - Fur is not the global regulator of iron uptake genes in Rhizobium leguminosarum. AB - Rhizobium leguminosarum fur mutants were unaffected in Fe-dependent regulation of several operons that specify different Fe uptake systems, yet cloned R. leguminosarum fur partially corrected an Escherichia coli fur mutant and R. leguminosarum Fur protein bound to canonical fur boxes. The lack of a phenotype in fur mutants is not due to functional redundancy with Irr, another member of the Fur superfamily found in the rhizobia, since irr fur double mutants are also unaffected in Fe-responsive regulation of several operons involved in Fe uptake. Neither Irr nor Fur is needed for symbiotic N(2) fixation on peas. As in Bradyrhizobium japonicum, irr mutants accumulated protoporphyrin IX. R. leguminosarum irr is not regulated by Fur and its Irr protein lacks the motif needed for haem-dependent post-translational modification that occurs in B. japonicum Irr. The similarities and differences in the Fur superfamily in the rhizobia and other Gram-negative bacteria are discussed. PMID- 12724398 TI - Distinct transcriptional pathways regulate basal and activated major histocompatibility complex class I expression. AB - Transcription of major histocompatibility complex (MHC) class I genes is regulated by both tissue-specific (basal) and hormone/cytokine (activated) mechanisms. Although promoter-proximal regulatory elements have been characterized extensively, the role of the core promoter in mediating regulation has been largely undefined. We report here that the class I core promoter consists of distinct elements that are differentially utilized in basal and activated transcription pathways. These pathways recruit distinct transcription factor complexes to the core promoter elements and target distinct transcription initiation sites. Class I transcription initiates at four major sites within the core promoter and is clustered in two distinct regions: "upstream" (-14 and -18) and "downstream" (+12 and +1). Basal transcription initiates predominantly from the upstream start site region and is completely dependent upon the general transcription factor TAF1 (TAF(II)250). Activated transcription initiates predominantly from the downstream region and is TAF1 (TAF(II)250) independent. USF1 augments transcription initiating through the upstream start sites and is dependent on TAF1 (TAF(II)250), a finding consistent with its role in regulating basal class I transcription. In contrast, transcription activated by the interferon mediator CIITA is independent of TAF1 (TAF(II)250) and focuses initiation on the downstream start sites. Thus, basal and activated transcriptions of an MHC class I gene target distinct core promoter domains, nucleate distinct transcription initiation complexes and initiate at distinct sites within the promoter. We propose that transcription initiation at the core promoter is a dynamic process in which the mechanisms of core promoter function differ depending on the cellular environment. PMID- 12724399 TI - Tax recruitment of CBP/p300, via the KIX domain, reveals a potent requirement for acetyltransferase activity that is chromatin dependent and histone tail independent. AB - Robust transcription of human T-cell leukemia virus type 1 (HTLV-1) genome requires the viral transactivator Tax. Although Tax has been previously shown to interact with the KIX domain of CBP/p300 in vitro, the precise functional relevance of this interaction remains unclear. Using two distinct approaches to interrupt the physical interaction between Tax and KIX, we find that Tax transactivation from chromatin templates is strongly dependent on CBP/p300 recruitment via the KIX domain. Additionally, we find that the primary functional contribution of CBP/p300 to Tax transactivation resides in the intrinsic acetyltransferase activity of the coactivators. These studies unexpectedly uncover a specific requirement for CBP/p300 acetyltransferase activity on chromatin templates assembled with nucleosomes lacking their amino-terminal tails. Together, these data indicate that the KIX domain of CBP/p300 is essential for targeting the acetyltransferase activity of the coactivator to the Tax-CREB (Tax/CREB) complex. Significantly, these observations reveal the presence of one or more CBP/p300 acetyltransferase targets that function specifically on chromatin templates, are independent of the histone tails, and are critical to Tax transactivation. PMID- 12724400 TI - Rad53 checkpoint kinase phosphorylation site preference identified in the Swi6 protein of Saccharomyces cerevisiae. AB - Rad53 of Saccharomyces cerevisiae is a checkpoint kinase whose structure and function are conserved among eukaryotes. When a cell detects damaged DNA, Rad53 activity is dramatically increased, which ultimately leads to changes in DNA replication, repair, and cell division. Despite its central role in checkpoint signaling, little is known about Rad53 substrates or substrate specificity. A number of proteins are implicated as Rad53 substrates; however, the evidence remains indirect. Previously, we have provided evidence that Swi6, a subunit of the Swi4/Swi6 late-G(1)-specific transcriptional activator, is a substrate of Rad53 in the G(1)/S DNA damage checkpoint. In the present study we identify Rad53 phosphorylation sites in Swi6 in vitro and demonstrate that at least one of them is targeted by Rad53 in vivo. Mutations in these phosphorylation sites in Swi6 shorten but do not eliminate the Rad53-dependent delay of the G(1)-to-S transition after DNA damage. We derive a consensus for Rad53 site preference at positions -2 and +2 (-2/+2) and identify its potential substrates in the yeast proteome. Finally, we present evidence that one of these candidates, the cohesin complex subunit Scc1 undergoes DNA damage-dependent phosphorylation, which is in part dependent on Rad53. PMID- 12724401 TI - A multiprotein nuclear complex connects Fanconi anemia and Bloom syndrome. AB - Bloom syndrome (BS) is a genetic disorder associated with dwarfism, immunodeficiency, reduced fertility, and an elevated risk of cancer. To investigate the mechanism of this disease, we isolated from human HeLa extracts three complexes containing the helicase defective in BS, BLM. Interestingly, one of the complexes, termed BRAFT, also contains five of the Fanconi anemia (FA) complementation group proteins (FA proteins). FA resembles BS in genomic instability and cancer predisposition, but most of its gene products have no known biochemical activity, and the molecular pathogenesis of the disease is poorly understood. BRAFT displays a DNA-unwinding activity, which requires the presence of BLM because complexes isolated from BLM-deficient cells lack such an activity. The complex also contains topoisomerase IIIalpha and replication protein A, proteins that are known to interact with BLM and could facilitate unwinding of DNA. We show that BLM complexes isolated from an FA cell line have a lower molecular mass. Our study provides the first biochemical characterization of a multiprotein FA complex and suggests a connection between the BLM and FA pathways of genomic maintenance. The findings that FA proteins are part of a DNA unwinding complex imply that FA proteins may participate in DNA repair. PMID- 12724402 TI - Protein-protein interaction between Fli-1 and GATA-1 mediates synergistic expression of megakaryocyte-specific genes through cooperative DNA binding. AB - Friend leukemia integration 1 (Fli-1) is a member of the Ets family of transcriptional activators that has been shown to be an important regulator during megakaryocytic differentiation. We undertook a two-hybrid screen of a K562 cDNA library to identify transcription factors that interacted with Fli-1 and were potential regulators of megakaryocyte development. Here we report the physical interaction of Fli-1 with GATA-1, a well-characterized, zinc finger transcription factor critical for both erythroid and megakaryocytic differentiation. We map the minimal domains required for the interaction and show that the zinc fingers of GATA-1 interact with the Ets domain of Fli-1. GATA-1 has previously been shown to interact with the Ets domain of the Fli-1-related protein PU.1, and the two proteins appear to inhibit each other's activity. In contrast, we demonstrate that GATA-1 and Fli-1 synergistically activate the megakaryocyte-specific promoters GPIX and GPIbalpha in transient transfections. Quantitative electrophoretic mobility shift assays using oligonucleotides derived from the GPIX promoter containing Ets and GATA binding motifs reveal that Fli-1 and GATA-1 exhibit cooperative DNA binding in which the binding of GATA-1 to DNA is increased approximately 26-fold in the presence of Fli-1 (from 4.2 to 0.16 nM), providing a mechanism for the observed transcriptional synergy. To test the effect on endogenous genes, we stably overexpressed Fli-1 in K562 cells, a line rich in GATA-1. Overexpression of Fli-1 induced the expression of the endogenous GPIX and GPIbalpha genes as measured by Northern blot and fluorescence-activated cell sorter analysis. This work suggests that Fli-1 and GATA-1 work together to activate the expression of genes associated with the terminal differentiation of megakaryocytes. PMID- 12724403 TI - The U1 snRNP base pairs with the 5' splice site within a penta-snRNP complex. AB - Recognition of the 5' splice site is an important step in mRNA splicing. To examine whether U1 approaches the 5' splice site as a solitary snRNP or as part of a multi-snRNP complex, we used a simplified in vitro system in which a short RNA containing the 5' splice site sequence served as a substrate in a binding reaction. This system allowed us to study the interactions of the snRNPs with the 5' splice site without the effect of other cis-regulatory elements of precursor mRNA. We found that in HeLa cell nuclear extracts, five spliceosomal snRNPs form a complex that specifically binds the 5' splice site through base pairing with the 5' end of U1. This system can accommodate RNA-RNA rearrangements in which U5 replaces U1 binding to the 5' splice site, a process that occurs naturally during the splicing reaction. The complex in which U1 and the 5' splice site are base paired sediments in the 200S fraction of a glycerol gradient together with all five spliceosomal snRNPs. This fraction is functional in mRNA spliceosome assembly when supplemented with soluble nuclear proteins. The results argue that U1 can bind the 5' splice site in a mammalian preassembled penta-snRNP complex. PMID- 12724404 TI - Identification and characterization of three new components of the mSin3A corepressor complex. AB - The mSin3A corepressor complex contains 7 to 10 tightly associated polypeptides and is utilized by many transcriptional repressors. Much of the corepressor function of mSin3A derives from associations with the histone deacetylases HDAC1 and HDAC2; however, the contributions of the other mSin3A-associated polypeptides remain largely unknown. We have purified an mSin3A complex from K562 erythroleukemia cells and identified three new mSin3A-associated proteins (SAP): SAP180, SAP130, and SAP45. SAP180 is 40% identical to a previously identified mSin3A-associated protein, RBP1. SAP45 is identical to mSDS3, the human ortholog of the SDS3p component of the Saccharomyces cerevisiae Sin3p-Rpd3p corepressor complex. SAP130 does not have detectable homology to other proteins. Coimmunoprecipitation and gel filtration data suggest that the new SAPs are, at the very least, components of the same mSin3A complex. Each new SAP repressed transcription when tethered to DNA. Furthermore, repression correlated with mSin3A binding, suggesting that the new SAPs are components of functional mSin3A corepressor complexes. SAP180 has two repression domains: a C-terminal domain, which interacts with the mSin3A-HDAC complex, and an N-terminal domain, which functions independently of mSin3A-HDAC. SAP130 has a repression domain at its C terminus that interacts with the mSin3A-HDAC complex and an N-terminal domain that probably mediates an interaction with a transcriptional activator. Together, our data suggest that these novel SAPs function in the assembly and/or enzymatic activity of the mSin3A complex or in mediating interactions between the mSin3A complex and other regulatory complexes. Finally, all three SAPs bind to the HDAC interaction domain (HID) of mSin3A, suggesting that the HID functions as the assembly interface for the mSin3A corepressor complex. PMID- 12724405 TI - Multiple mechanistically distinct functions of SAGA at the PHO5 promoter. AB - Our previous studies have shown that the rate of chromatin remodeling and consequently the rate of PHO5 activation are strongly decreased in the absence of Gcn5 histone acetyltransferase activity. Using chromatin immunoprecipitation, we demonstrate that SAGA is physically recruited to the PHO5 promoter. Recruitment is dependent on the specific activator Pho4 and occurs only under inducing conditions. Spt3, another subunit of SAGA, also plays a role in PHO5 activation but has a function that is completely different from that of Gcn5. An SPT3 deletion severely compromises the PHO5 promoter and reduces the extent of transcriptional activation by diminishing the binding of the TATA binding protein to the promoter without, however, affecting the rate or the extent of chromatin remodeling. A gcn5 spt3 double mutant shows a synthetic phenotype almost as severe as that observed for an spt7 or spt20 mutant. The latter two mutations are known to prevent the assembly of the complex and consequently lead to the loss of all SAGA functions. The absence of the Ada2 subunit causes a strong delay in chromatin remodeling and promoter activation that closely resembles the delay observed in the absence of Gcn5. A deletion of only the Ada2 SANT domain has exactly the same effect, strongly suggesting that Ada2 controls Gcn5 activity by virtue of its SANT domain. Finally, the Gcn5 bromodomain also contributes to but is not essential for Gcn5 function at the PHO5 promoter. Taken together, the results provide a detailed and differentiated description of the role of SAGA as a coactivator at the PHO5 promoter. PMID- 12724406 TI - Human BAG-1 proteins bind to the cellular stress response protein GADD34 and interfere with GADD34 functions. AB - The cellular stress response protein GADD34 mediates growth arrest and apoptosis in response to DNA damage, negative growth signals, and protein malfolding. GADD34 binds to protein phosphatase PP1 and can attenuate the translational elongation of key transcriptional factors through dephosphorylation of eukaryotic initiation factor 2alpha (eIF2alpha). Recently, we reported the involvement of human SNF5/INI1 (hSNF5/INI1) protein in the functions of GADD34 and showed that hSNF5/INI1 binds GADD34 and stimulates the bound PP1 phosphatase activity. To better understand the regulatory and functional mechanisms of GADD34, we undertook a yeast two-hybrid screen with full-length GADD34 as bait in order to identify additional protein partners of GADD34. We report here that human cochaperone protein BAG-1 interacts with GADD34 in vitro and in SW480 cells treated with the proteasome inhibitor z-LLL-B to induce apoptosis. Two other proteins, Hsp70/Hsc70 and PP1, associate reversibly with the GADD34-BAG-1 complex, and their dissociation is promoted by ATP. BAG-1 negatively modulates GADD34-bound PP1 activity, and the expression of BAG-1 isoforms can also mask GADD34-mediated inhibition of colony formation and suppression of transcription. Our findings suggest that BAG-1 may function to suppress the GADD34-mediated cellular stress response and support a role for BAG-1 in the survival of cells undergoing stress. PMID- 12724407 TI - The mechanism of Mus81-Mms4 cleavage site selection distinguishes it from the homologous endonuclease Rad1-Rad10. AB - Mus81-Mms4 and Rad1-Rad10 are homologous structure-specific endonucleases that cleave 3' branches from distinct substrates and are required for replication fork stability and nucleotide excision repair, respectively, in the yeast Saccharomyces cerevisiae. We explored the basis of this biochemical and genetic specificity. The Mus81-Mms4 cleavage site, a nick 5 nucleotides (nt) 5' of the flap, is determined not by the branch point, like Rad1-Rad10, but by the 5' end of the DNA strand at the flap junction. As a result, the endonucleases show inverse substrate specificity; substrates lacking a 5' end within 4 nt of the flap are cleaved poorly by Mus81-Mms4 but are cleaved well by Rad1-10. Genetically, we show that both mus81 and sgs1 mutants are sensitive to camptothecin-induced DNA damage. Further, mus81 sgs1 synthetic lethality requires homologous recombination, as does suppression of mutant phenotypes by RusA expression. These data are most easily explained by a model in which the in vivo substrate of Mus81-Mms4 and Sgs1-Top3 is a 3' flap recombination intermediate downstream of replication fork collapse. PMID- 12724408 TI - Two ubiquitin-conjugating enzymes, UbcP1/Ubc4 and UbcP4/Ubc11, have distinct functions for ubiquitination of mitotic cyclin. AB - Cell cycle events are regulated by sequential activation and inactivation of Cdk kinases. Mitotic exit is accomplished by the inactivation of mitotic Cdk kinase, which is mainly achieved by degradation of cyclins. The ubiquitin-proteasome system is involved in this process, requiring APC/C (anaphase-promoting complex/cyclosome) as a ubiquitin ligase. In Xenopus and clam oocytes, the ubiquitin-conjugating enzymes that function with APC/C have been identified as two proteins, UBC4 and UBCx/E2-C. Previously we reported that the fission yeast ubiquitin-conjugating enzyme UbcP4/Ubc11, a homologue of UBCx/E2-C, is required for mitotic transition. Here we show that the other fission yeast ubiquitin conjugating enzyme, UbcP1/Ubc4, which is homologous to UBC4, is also required for mitotic transition in the same manner as UbcP4/Ubc11. Both ubiquitin-conjugating enzymes are essential for cell division and directly required for the degradation of mitotic cyclin Cdc13. They function nonredundantly in the ubiquitination of CDC13 because a defect in ubcP1/ubc4+ cannot be suppressed by high expression of UbcP4/Ubc11 and a defect in ubcP4/ubc11+ cannot be suppressed by high expression of UbcP1/Ubc4. In vivo analysis of the ubiquitinated state of Cdc13 shows that the ubiquitin chains on Cdc13 were short in ubcP1/ubc4 mutant cells while ubiquitinated Cdc13 was totally reduced in ubcP4/ubc11 mutant cells. Taken together, these results indicate that the two ubiquitin-conjugating enzymes play distinct and essential roles in the degradation of mitotic cyclin Cdc13, with the UbcP4/Ubc11-pathway initiating ubiquitination of Cdc13 and the UbcP1/Ubc4-pathway elongating the short ubiquitin chains on Cdc13. PMID- 12724409 TI - A constitutive decay element promotes tumor necrosis factor alpha mRNA degradation via an AU-rich element-independent pathway. AB - Tumor necrosis factor alpha (TNF-alpha) expression is regulated by transcriptional as well as posttranscriptional mechanisms, the latter including the control of mRNA decay through an AU-rich element (ARE) in the 3' untranslated region (UTR). Using two mutant cell lines deficient for ARE-mediated mRNA decay, we provide evidence for a second element, the constitutive decay element (CDE), which is also located in the 3' UTR of TNF-alpha. In stably transfected RAW 264.7 macrophages stimulated with lipopolysaccharide (LPS), the CDE continues to target a reporter transcript for rapid decay, whereas ARE-mediated decay is blocked. Similarly, the activation of p38 kinase and phosphatidylinositol 3-kinase in NIH 3T3 cells inhibits ARE-mediated but not CDE-mediated mRNA decay. The CDE was mapped to an 80-nucleotide (nt) segment downstream of the ARE, and point mutation analysis identified within the CDE a conserved sequence of 15 nt that is required for decay activity. We propose that the CDE represses TNF-alpha expression by maintaining the mRNA short-lived, thereby preventing excessive induction of TNF alpha after LPS stimulation. Thus, CDE-mediated mRNA decay is likely to be an important mechanism limiting LPS-induced pathologic processes. PMID- 12724410 TI - Chromosomal elements regulate gene activity and chromatin structure of the human serpin gene cluster at 14q32.1. AB - The human serine protease inhibitor (serpin) gene cluster at 14q32.1 contains a number of genes that are specifically expressed in hepatic cells. Cell-specific enhancers have been identified in several of these genes, but elements involved in locus-wide gene and chromatin control have yet to be defined. To identify regulatory elements in this region, we prepared a series of mutant chromosomal alleles by homologous recombination and transferred the specifically modified human chromosomes to hepatic cells for functional tests. We report that deletion of an 8-kb DNA segment upstream of the human alpha1-antitrypsin gene yields a mutant serpin allele that fails to be activated in hepatic cells. Within this region, a 2.3-kb DNA segment between kb -8.1 and -5.8 contains a previously unrecognized control region that is required not only for serpin gene activation but also for chromatin remodeling of the entire locus. PMID- 12724411 TI - Functional relation among RecQ family helicases RecQL1, RecQL5, and BLM in cell growth and sister chromatid exchange formation. AB - Human RECQL1 and RECQL5 belong to the RecQ family that includes Bloom syndrome, Werner syndrome, and Rothmund-Thomson syndrome causative genes. Cells derived from individuals suffering from these syndromes show significant levels of genomic instability. However, neither RECQL1 nor RECQL5 has been related to a disease, and nothing is known about the functions of RecQL1 and RecQL5. We generated here RECQL1(-/-), RECQL5(-/-), RECQL1(-/-)/RECQL5(-/-), RECQL1(-/ )/BLM(-/-), and RECQL5(-/-)/BLM(-/-) cells from chicken B-lymphocyte line DT40 cells. Although BLM(-/-) DT40 cells showed a slow-growth phenotype, a higher sensitivity to methyl methanesulfonate than the wild type, and an approximately 10-fold increase in the frequency of sister chromatid exchange (SCE) compared to wild-type cells, RECQL1(-/-), RECQL5(-/-), and RECQL1(-/-)/RECQL5(-/-) cells showed no significant difference from the wild-type cells in growth, sensitivity to DNA-damaging agents, and the frequency of SCE. However, both RECQL1(-/-)/BLM( /-) and RECQL5(-/-)/BLM(-/-) cells grew more slowly than BLM(-/-) cells because of the increase in the population of dead cells, indicating that RecQL1 and RecQL5 are somehow involved in cell viability under the BLM function-impaired condition. Surprisingly, RECQL5(-/-)/BLM(-/-) cells showed a higher frequency of SCE than BLM(-/-) cells, indicating that RecQL5 suppresses SCE under the BLM function-impaired condition. PMID- 12724412 TI - Replication of the chicken beta-globin locus: early-firing origins at the 5' HS4 insulator and the rho- and betaA-globin genes show opposite epigenetic modifications. AB - Chromatin structure is believed to exert a strong effect on replication origin function. We have studied the replication of the chicken beta-globin locus, whose chromatin structure has been extensively characterized. This locus is delimited by hypersensitive sites (HSs) that mark the position of insulator elements. A stretch of condensed chromatin and another HS separate the beta-globin domain from an adjacent folate receptor (FR) gene. We demonstrate here that in erythroid cells that express the FR but not the globin genes, replication initiates at four sites within the beta-globin domain, one at the 5' HS4 insulator and the other three near the rho- and beta(A)-globin genes. Three origins consist of G+C-rich sequences enriched in CpG dinucleotides. The fourth origin is A+T rich. Together with previous work, these data reveal that the insulator origin has unmethylated CpGs, hyperacetylated histones H3 and H4, and lysine 4-methylated histone H3. In contrast, opposite modifications are observed at the other G+C-rich origins. We also show that the whole region, including the stretch of condensed chromatin, replicates early in S phase in these cells. Therefore, different early-firing origins within the same locus may have opposite patterns of epigenetic modifications. The role of insulator elements in DNA replication is discussed. PMID- 12724413 TI - Human gene targeting by adeno-associated virus vectors is enhanced by DNA double strand breaks. AB - The use of adeno-associated virus (AAV) to package gene-targeting vectors as single-stranded linear molecules has led to significant improvements in mammalian gene-targeting frequencies. However, the molecular basis for the high targeting frequencies obtained is poorly understood, and there could be important mechanistic differences between AAV-mediated gene targeting and conventional gene targeting with transfected double-stranded DNA constructs. Conventional gene targeting is thought to occur by the double-strand break (DSB) model of homologous recombination, as this can explain the higher targeting frequencies observed when DSBs are present in the targeting construct or target locus. Here we compare AAV-mediated gene-targeting frequencies in the presence and absence of induced target site DSBs. Retroviral vectors were used to introduce a mutant lacZ gene containing an I-SceI cleavage site and to efficiently deliver the I-SceI endonuclease, allowing us to carry out these studies with normal and transformed human cells. Creation of DSBs by I-SceI increased AAV-mediated gene-targeting frequencies 60- to 100-fold and resulted in a precise correction of the mutant lacZ reporter gene. These experiments demonstrate that AAV-mediated gene targeting can result in repair of a DNA DSB and that this form of gene targeting exhibits fundamental similarities to conventional gene targeting. In addition, our findings suggest that the selective creation of DSBs by using viral delivery systems can increase gene-targeting frequencies in scientific and therapeutic applications. PMID- 12724414 TI - Efficient gene targeting mediated by adeno-associated virus and DNA double-strand breaks. AB - Gene targeting is the in situ manipulation of the sequence of an endogenous gene by the introduction of homologous exogenous DNA. Presently, the rate of gene targeting is too low for it to be broadly used in mammalian somatic cell genetics or to cure genetic diseases. Recently, it has been demonstrated that infection with recombinant adeno-associated virus (rAAV) vectors can mediate gene targeting in somatic cells, but the mechanism is unclear. This paper explores the balance between random integration and gene targeting with rAAV. Both random integration and spontaneous gene targeting are dependent on the multiplicity of infection (MOI) of rAAV. It has previously been shown that the introduction of a DNA double stranded break (DSB) in a target gene can stimulate gene targeting by several thousand-fold in somatic cells. Creation of a DSB stimulates the frequency of rAAV-mediated gene targeting by over 100-fold, suggesting that the mechanism of rAAV-mediated gene targeting involves, at least in part, the repair of DSBs by homologous recombination. Absolute gene targeting frequencies reach 0.8% with a dual vector system in which one rAAV vector provides a gene targeting substrate and a second vector expresses the nuclease that creates a DSB in the target gene. The frequencies of gene targeting that we achieved with relatively low MOIs suggest that combining rAAV vectors with DSBs is a promising strategy to broaden the application of gene targeting. PMID- 12724415 TI - Direct involvement of HERV-W Env glycoprotein in human trophoblast cell fusion and differentiation. AB - We recently demonstrated that the product of the HERV-W env gene, a retroviral envelope protein also dubbed syncytin, is a highly fusogenic membrane glycoprotein inducing the formation of syncytia on interaction with the type D mammalian retrovirus receptor. In addition, the detection of HERV-W Env protein (Env-W) expression in placental tissue sections led us to propose a role for this fusogenic glycoprotein in placenta formation. To evaluate this hypothesis, we analyzed the involvement of Env-W in the differentiation of primary cultures of human villous cytotrophoblasts that spontaneously differentiate by cell fusion into syncytiotrophoblasts in vitro. First, we observed that HERV-W env mRNA and glycoprotein expression are colinear with primary cytotrophoblast differentiation and with expression of human chorionic gonadotropin (hCG), a marker of syncytiotrophoblast formation. Second, we observed that in vitro stimulation of trophoblast cell fusion and differentiation by cyclic AMP is also associated with a concomitant increase in HERV-W env and hCG mRNA and protein expression. Finally, by using specific antisense oligonucleotides, we demonstrated that inhibition of Env-W protein expression leads to a decrease of trophoblast fusion and differentiation, with the secretion of hCG in culture medium of antisense oligonucleotide-treated cells being decreased by fivefold. Taken together, these results strongly support a direct role for Env-W in human trophoblast cell fusion and differentiation. PMID- 12724416 TI - Mice lacking the giant protocadherin mFAT1 exhibit renal slit junction abnormalities and a partially penetrant cyclopia and anophthalmia phenotype. AB - While roles in adhesion and morphogenesis have been documented for classical cadherins, the nonclassical cadherins are much less well understood. Here we have examined the functions of the giant protocadherin FAT by generating a transgenic mouse lacking mFAT1. These mice exhibit perinatal lethality, most probably caused by loss of the renal glomerular slit junctions and fusion of glomerular epithelial cell processes (podocytes). In addition, some mFAT1(-/-) mice show defects in forebrain development (holoprosencephaly) and failure of eye development (anophthalmia). In contrast to Drosophila, where FAT acts as a tumor suppressor gene, we found no evidence for abnormalities of proliferation in two tissues (skin and central nervous system [CNS]) containing stem and precursor cell populations and in which FAT is expressed strongly. Our results confirm a necessary role for FAT1 in the modified adhesion junctions of the renal glomerular epithelial cell and reveal hitherto unsuspected roles for FAT1 in CNS development. PMID- 12724417 TI - Activating signal cointegrator 2 required for liver lipid metabolism mediated by liver X receptors in mice. AB - Activating signal cointegrator 2 (ASC-2), a cancer-amplified transcriptional coactivator of nuclear receptors and many other transcription factors, contains two LXXLL-type nuclear receptor interaction domains. Interestingly, the second LXXLL motif is highly specific to the liver X receptors (LXRs). In cotransfection, DN2, an ASC-2 fragment encompassing this motif, exerts a potent dominant-negative effect on transactivation by LXRs, which is rescued by ectopic coexpression of the full-length ASC-2 but not by other LXXLL-type coactivators, such as SRC-1 and TRAP220. In contrast, DN2/m, in which the LXXLL motif is mutated to LXXAA to abolish the interactions with LXRs, is without any effect. Accordingly, expression of DN2, but not DN2/m, in transgenic mice results in phenotypes that are highly homologous to those previously observed with LXRalpha( /-) mice, including a rapid accumulation of large amounts of cholesterol and down regulation of the known lipid-metabolizing target genes of LXRalpha in the liver upon being fed a high-cholesterol diet. These results identify ASC-2 as a physiologically important transcriptional coactivator of LXRs and demonstrate its pivotal role in the liver lipid metabolism. PMID- 12724418 TI - Cardiac p300 is involved in myocyte growth with decompensated heart failure. AB - A variety of stresses on the heart initiate a number of subcellular signaling pathways, which finally reach the nuclei of cardiac myocytes and cause myocyte hypertrophy with heart failure. However, common nuclear pathways that lead to this state are unknown. A zinc finger protein, GATA-4, is one of the transcription factors that mediate changes in gene expression during myocardial cell hypertrophy. p300 not only acts as a transcriptional coactivator of GATA-4, but also possesses an intrinsic histone acetyltransferase activity. In primary cardiac myocytes derived from neonatal rats, we show that stimulation with phenylephrine increased an acetylated form of GATA-4 and its DNA-binding activity, as well as expression of p300. A dominant-negative mutant of p300 suppressed phenylephrine-induced nuclear acetylation, activation of GATA-4 dependent endothelin-1 promoters, and hypertrophic responses, such as increase in cell size and sarcomere organization. In sharp contrast to the activation of cardiac MEK-1, which phosphorylates GATA-4 and causes compensated hypertrophy in vivo, p300-mediated acetylation of mouse cardiac nuclear proteins, including GATA 4, results in marked eccentric dilatation and systolic dysfunction. These findings suggest that p300-mediated nuclear acetylation plays a critical role in the development of myocyte hypertrophy and represents a pathway that leads to decompensated heart failure. PMID- 12724419 TI - Defective gene expression, S phase progression, and maturation during hematopoiesis in E2F1/E2F2 mutant mice. AB - E2F plays critical roles in cell cycle progression by regulating the expression of genes involved in nucleotide synthesis, DNA replication, and cell cycle control. We show that the combined loss of E2F1 and E2F2 in mice leads to profound cell-autonomous defects in the hematopoietic development of multiple cell lineages. E2F2 mutant mice show erythroid maturation defects that are comparable with those observed in patients with megaloblastic anemia. Importantly, hematopoietic defects observed in E2F1/E2F2 double-knockout (DKO) mice appear to result from impeded S phase progression in hematopoietic progenitor cells. During DKO B-cell maturation, differentiation beyond the large pre-BII-cell stage is defective, presumably due to failed cell cycle exit, and the cells undergo apoptosis. However, apoptosis appears to be the consequence of failed maturation, not the cause. Despite the accumulation of hematopoietic progenitor cells in S phase, the combined loss of E2F1 and E2F2 results in significantly decreased expression and activities of several E2F target genes including cyclin A2. Our results indicate specific roles for E2F1 and E2F2 in the induction of E2F target genes, which contribute to efficient expansion and maturation of hematopoietic progenitor cells. Thus, E2F1 and E2F2 play essential and redundant roles in the proper coordination of cell cycle progression with differentiation which is necessary for efficient hematopoiesis. PMID- 12724420 TI - FAP-1 association with Fas (Apo-1) inhibits Fas expression on the cell surface. AB - As revealed by intracellular pools of nonactive Fas (Apo-1), export of Fas to the cell surface is often impaired in human tumors, thereby inactivating Fas ligand mediated apoptosis. Here, we demonstrate that association with Fas-associated phosphatase 1 (FAP-1) attenuates Fas export to the cell surface. Forced expression of FAP-1 reduces cell surface Fas levels and increases the intracellular pool of Fas within the cytoskeleton network. Conversely, expression of dominant-negative forms of FAP-1, or inhibition of FAP-1 expression by short interfering RNA, efficiently up-regulates surface expression of Fas. Inhibition of Fas surface expression by FAP-1 depends on its association with the C terminus of Fas. Mutation within amino acid 275 results in decreased association with FAP 1 and greater export of Fas to the cell surface in melanomas, normal fibroblasts, or Fas null cells. Identifying the role of FAP-1 in binding to, and consequently inhibition of, Fas export to the cell surface provides novel insight into the mechanism underlying the regulation of Fas trafficking, which is commonly impaired in advanced tumors with FAP-1 overexpression. PMID- 12724421 TI - Nonmuscle myosin promotes cytoplasmic localization of PBX. AB - In the absence of MEIS family proteins, two mechanisms are known to restrict the PBX family of homeodomain (HD) transcription factors to the cytoplasm. First, PBX is actively exported from the nucleus via a CRM1-dependent pathway. Second, nuclear localization signals (NLSs) within the PBX HD are masked by intramolecular contacts. In a screen to identify additional proteins directing PBX subcellular localization, we identified a fragment of murine nonmuscle myosin II heavy chain B (NMHCB). The interaction of NMHCB with PBX was verified by coimmunoprecipitation, and immunofluorescence staining revealed colocalization of NMHCB with cytoplasmic PBX in the mouse embryo distal limb bud. The interaction domain in PBX mapped to a conserved PBC-B region harboring a potential coiled coil structure. In support of the cytoplasmic retention function, the NMHCB fragment competes with MEIS1A to redirect PBX, and the fly PBX homologue EXD, to the cytoplasm of mammalian and insect cells. Interestingly, MEIS1A also localizes to the cytoplasm in the presence of the NMHCB fragment. These activities are largely independent of nuclear export. We show further that the subcellular localization of EXD is deregulated in Drosophila zipper mutants that are depleted of nonmuscle myosin heavy chain. This study reveals a novel and evolutionarily conserved mechanism controlling the subcellular distribution of PBX and EXD proteins. PMID- 12724422 TI - Modeling del(17)(p11.2p11.2) and dup(17)(p11.2p11.2) contiguous gene syndromes by chromosome engineering in mice: phenotypic consequences of gene dosage imbalance. AB - Contiguous gene syndromes (CGS) are a group of disorders associated with chromosomal rearrangements of which the phenotype is thought to result from altered copy numbers of physically linked dosage-sensitive genes. Smith-Magenis syndrome (SMS) is a CGS associated with a deletion within band p11.2 of chromosome 17. Recently, patients harboring the predicted reciprocal duplication product [dup(17)(p11.2p11.2)] have been described as having a relatively mild phenotype. By chromosomal engineering, we created rearranged chromosomes carrying the deletion [Df(11)17] or duplication [Dp(11)17] of the syntenic region on mouse chromosome 11 that spans the genomic interval commonly deleted in SMS patients. Df(11)17/+ mice exhibit craniofacial abnormalities, seizures, marked obesity, and male-specific reduced fertility. Dp(11)17/+ animals are underweight and do not have seizures, craniofacial abnormalities, or reduced fertility. Examination of Df(11)17/Dp(11)17 animals suggests that most of the observed phenotypes result from gene dosage effects. Our murine models represent a powerful tool to analyze the consequences of gene dosage imbalance in this genomic interval and to investigate the molecular genetic bases of both SMS and dup(17)(p11.2p11.2). PMID- 12724424 TI - The cyclin E/Cdk2 substrate and Cajal body component p220(NPAT) activates histone transcription through a novel LisH-like domain. AB - p220(NPAT) is a substrate of cyclin E/Cdk2 that localizes in nuclear organelles called Cajal bodies in a cell cycle-regulated manner. In normal diploid fibroblasts, p220 is concentrated in two Cajal bodies tethered to histone gene clusters at chromosome 6p21 during G(1), S, and G(2) phases and two additional Cajal bodies tethered to histone genes at 1q21 during S, and G(2) phases. Overexpression of p220 in U2OS cells can promote the G(1)/S transition and can also promote transcription from histone H2B and H4 luciferase reporter constructs. How p220 expression induces these activities and whether the two activities are related are unknown. In this study, we developed a "lox-scanning" mutagenesis approach to identify functional domains in p220. We identified two distinct functional regions of p220. The C-terminal half of the protein contains multiple elements that are required for its ability to induce S phase in transfected cells. In contrast, sequences at the N terminus appear to be critical for activation of histone H4 and H2B reporter constructs. We identified an approximately 30-amino-acid motif at the N terminus of p220 that has the characteristics of a LisH motif. LisH motifs are found in a large number of proteins in the database but are of unknown function. Point mutations in conserved residues in the LisH motif of p220 block histone H4 transcriptional activity without affecting localization in Cajal bodies or phosphorylation on Cdk2 phosphorylation sites. These studies indicate that the ability of p220 to promote S phase is independent of its ability to promote histone H4 transcription and suggests that p220 may link cyclin E/Cdk2 to multiple independent downstream functions. PMID- 12724423 TI - Constitutive E2F1 overexpression delays endochondral bone formation by inhibiting chondrocyte differentiation. AB - Longitudinal bone growth results from endochondral ossification, a process that requires proliferation and differentiation of chondrocytes. It has been shown that proper endochondral bone formation is critically dependent on the retinoblastoma family members p107 and p130. However, the precise functional roles played by individual E2F proteins remain poorly understood. Using both constitutive and conditional E2F1 transgenic mice, we show that ubiquitous transgene-driven expression of E2F1 during embryonic development results in a dwarf phenotype and significantly reduced postnatal viability. Overexpression of E2F1 disturbs chondrocyte maturation, resulting in delayed endochondral ossification, which is characterized by reduced hypertrophic zones and disorganized growth plates. Employing the chondrogenic cell line ATDC5, we investigated the effects of enforced E2F expression on the different phases of chondrocyte maturation that are normally required for endochondral ossification. Ectopic E2F1 expression strongly inhibits early- and late-phase differentiation of ATDC5 cells, accompanied by diminished cartilage nodule formation as well as decreased type II collagen, type X collagen, and aggrecan gene expression. In contrast, overexpression of E2F2 or E2F3a results in only a marginal delay of chondrocyte maturation, and increased E2F4 levels have no effect. These data are consistent with the notion that E2F1 is a regulator of chondrocyte differentiation. PMID- 12724425 TI - Telomeric position effect variegation in Saccharomyces cerevisiae by Caenorhabditis elegans linker histones suggests a mechanistic connection between germ line and telomeric silencing. AB - Linker histones are nonessential for the life of single-celled eukaryotes. Linker histones, however, can be important components of specific developmental programs in multicellular animals and plants. For Caenorhabditis elegans a single linker histone variant (H1.1) is essential in a chromatin silencing process which is crucial for the proliferation and differentiation of the hermaphrodite germ line. In this study we analyzed the whole linker histone complement of C. elegans by telomeric position effect variegation in budding yeast. In this assay an indicator gene (URA3) placed close to the repressive telomeric chromatin structure is subject to epigenetically inherited gene inactivation. Just one out of seven C. elegans linker histones (H1.1) was able to enhance the telomeric position effect in budding yeast. Since these results reflect the biological function of H1.1 in C. elegans, we suggest that chromatin silencing in C. elegans is governed by molecular mechanisms related to the telomere-dependent silencing in budding yeast. We confirmed this hypothesis by testing C. elegans homologs of three yeast genes which are established modifiers of the yeast telomeric chromatin structure (SIR2, SET1, and RAD17) for their influence on repeat dependent transgene silencing for C. elegans. PMID- 12724427 TI - DIPEx: fresh insights for medical practice. PMID- 12724426 TI - Role for the fission yeast RecQ helicase in DNA repair in G2. AB - Members of the RecQ helicase subfamily are mutated in several human genomic instability syndromes, such as Bloom, Werner, and Rothmund-Thomson syndromes. We show that Rqh1, the single Schizosaccharomyces pombe homologue, is a 3'-to-5' helicase and exists with Top3 in a high-molecular-weight complex. top3 deletion is inviable, and this is suppressed by concomitant loss of rqh1 helicase activity or loss of recombination functions. This is consistent with RecQ helicases in other systems. By using epistasis analysis of the UV radiation sensitivity and by analyzing the kinetics of Rhp51 (Rad51 homologue), Rqh1, and Top3 focus formation in response to UV in synchronized cells, we identify the first evidence of a function for Rqh1 and Top3 in the repair of UV-induced DNA damage in G(2). Our data provide evidence that Rqh1 functions after Rad51 focus formation during DNA repair. We also identify a function for Rqh1 upstream of recombination in an Rhp18-dependent (Rad18 homologue) pathway. The model that these data allow us to propose helps to reconcile different interpretations of RecQ family helicase function that have arisen between work based on the S. pombe system and models based on studies of Saccharomyces cerevisiae SGS1 suggesting that RecQ helicases act before Rad51. PMID- 12724428 TI - Violence--a noxious cocktail of genes and the environment. PMID- 12724429 TI - Oxygen or air for palliation of breathlessness in advanced cancer. PMID- 12724430 TI - Patients' memory for medical information. PMID- 12724432 TI - Hypothermia and acute pancreatitis: myth or reality? AB - Among the rarer causes of acute pancreatitis listed in surgical texts is hypothermia. To assess the evidence for cause and effect, we questioned selected consultants about their experience and examined the case-notes of patients admitted with hypothermia. The 31 consultants who returned our questionnaire (69% response rate; 317 consultant-years' experience) could recall only 5 cases of pancreatitis associated with hypothermia, in 2 of which other aetiological factors were judged primary. In case-notes for 100 months of emergency admissions at a single hospital we identified 310 patients with hypothermia and 1153 with acute pancreatitis; none had the dual diagnosis. Of the hypothermic patients, none had abdominal pain typical of acute pancreatitis. In 43 serum amylase was measured because the patient was unable to give a full history and in 2 of these the enzyme was slightly raised; both had experienced a cerebrovascular accident, which is a known cause of hyperamylasaemia. Considered alongside the weak evidence from previous studies, these findings offer negligible support for the idea that hypothermia is a clinically relevant risk factor for acute pancreatitis. PMID- 12724431 TI - Medically unexplained symptoms: exacerbating factors in the doctor-patient encounter. PMID- 12724433 TI - Nasal reconstruction in the Yemen with the Converse scalping flap. PMID- 12724434 TI - Serotonin syndrome after single doses of co-amoxiclav during treatment with venlafaxine. PMID- 12724435 TI - Two cases of pregnancy with coarctation of the aorta. PMID- 12724436 TI - A painful swollen calf. PMID- 12724437 TI - Isolated septic arthritis: meningococcal infection. PMID- 12724438 TI - Sustained remission of metastatic Merkel cell carcinoma with treatment of HIV infection. PMID- 12724439 TI - Rheumatoid hydrocele. PMID- 12724440 TI - 20 Hanover Square, London W1: first home of the Royal Society of Medicine. PMID- 12724441 TI - A five-year pregnancy. PMID- 12724442 TI - Legal considerations of clinical guidelines. PMID- 12724443 TI - Autonomy of the pregnant woman. PMID- 12724444 TI - Legal considerations of clinical guidelines. PMID- 12724445 TI - Myths and mandrakes. PMID- 12724446 TI - Myths and mandrakes. PMID- 12724447 TI - Atypical mycobacterial infections. PMID- 12724448 TI - Supporting individuals with disabling multiple sclerosis. PMID- 12724449 TI - Michelangelo and medicine. PMID- 12724450 TI - Lymphocytic hypophysitis. PMID- 12724451 TI - Doctors' knowledge about consent and capacity. PMID- 12724452 TI - Functional magnetic resonance imaging: application to posttraumatic stress disorder. PMID- 12724453 TI - A critical review of memory stimulation programs in Alzheimer's disease. AB - The authors describe the memory stimulation programs used in the treatment of Alzheimer's disease (AD) and review their efficacy. Visual imagery, errorless learning, dyadic approaches, spaced retrieval techniques, encoding specificity with cognitive support at retrieval, and external memory aids were the memory stimulation programs used alone or in combination in AD. Preliminary evidence suggests that the errorless learning, spaced retrieval, and vanishing cues techniques and the dyadic approach, used alone or in combination, are efficacious in stimulating memory in patients with AD. PMID- 12724454 TI - Electrophysiological aberrations in borderline personality disorder: state of the evidence. AB - Electrophysiological technology is noninvasive and relatively inexpensive. In order to assess the usefulness of this technology in probing the pathophysiology of borderline personality disorder (BPD), we reviewed the literature in which an electrophysiological modality was used to examine BPD. Twenty-two articles were identified, from which diagnostic criteria and data on comorbidity and control groups were extracted. Although the majority of studies pointed to a high prevalence of electrophysiological aberrations in patients, very few studies had adequate control groups and adequate evaluation of comorbidity. We conclude that although the existing literature reflects a preliminary stage of the field, it suggests that electrophysiological investigations may be useful in investigating BPD. PMID- 12724455 TI - Clinical correlates of aggressive behavior after traumatic brain injury. AB - The authors assessed aggressive behavior in 89 patients with traumatic brain injury (TBI) and 26 patients with multiple trauma but without TBI using a quantitative scale (the Overt Aggression Scale) and examined its clinical correlates. Aggressive behavior was found in 33.7% of TBI patients and 11.5% of patients without TBI during the first 6 months after injury. Aggressive behavior was significantly associated with the presence of major depression, frontal lobe lesions, poor premorbid social functioning, and a history of alcohol and substance abuse. Interventions aimed at treatment of depression and substance abuse and enhancing social support may help reduce the severity of this disruptive behavior. PMID- 12724456 TI - Nulliparity and late menopause are associated with decreased cognitive decline. AB - Changes in Mini-Mental State Examination (MMSE) scores were examined over a median of 12.8 years in a population of 361 community-dwelling postmenopausal women who had never received estrogen replacement therapy. In a linear regression model that took into account age, education, race, surgical versus natural menopause, use of birth control pills, and MMSE score at baseline, it was found that nulliparous women and women who went through menopause later in life had significantly less cognitive decline. These results suggest that greater lifetime exposure to endogenous estrogen may be associated with less age-related cognitive decline. PMID- 12724458 TI - Diagnostic utility of visual evoked potential changes in Alzheimer's disease. AB - Previous studies have consistently found a selective delay of the P2 flash visual evoked potential (VEP) component among groups of patients with Alzheimer's dementia (AD) compared with control groups. Several authors have termed the selective P2 delay a "marker" for AD and have called for its use in clinical diagnosis. This study examined the diagnostic utility of the selective P2 delay in a retrospective sample of 45 AD patients and 60 age-equivalent healthy control subjects. Although significant between-group differences were found, classification accuracies for individual patients and controls were too low for the P2 delay to contribute meaningfully to clinical diagnosis. PMID- 12724457 TI - Magical ideation modulates spatial behavior. AB - Previous research has found that animals as well as persons with psychotic disorders preferentially orient away from the cerebral hemisphere with the more active dopamine system. This study investigated the modulation of spatial behavior by a mode of thinking reminiscent of the positive symptoms of psychosis. In a non-treatment-seeking sample of healthy volunteers (20 women and 16 men), the authors assessed the lateral biases in turning and veering behavior and in line bisection as a function of their magical ideation, that is, a mild form of schizotypy. Across tasks, pronounced magical ideation was associated with reduced right-sided orientation preferences. This finding suggests a relative hyperdopaminergia of the right hemisphere as the biological basis of magical ideation. PMID- 12724459 TI - Frequency and characteristics of anxiety among patients with Alzheimer's disease and related dementias. AB - The purpose of this study was to assess the cross-sectional prevalence and characteristics of anxiety among patients with Alzheimer's disease (AD), as compared with patients with frontotemporal dementia (FTD), patients with vascular dementia (VaD), and normal control subjects. The authors used the anxiety subscale of the Neuropsychiatric Inventory (NPI), an instrument with established reliability and validity, to compare patients. Patients were identified in a query of the UCLA Alzheimer's Disease Center database and included 115 patients with probable AD, 43 patients with VaD, 33 patients with FTD, and 40 normal, elderly control subjects. Descriptive statistics were generated, and partial correlations, controlling for Mini-Mental State Examination (MMSE) score, were performed between the anxiety subscale and other behavioral features as measured by the NPI and the Functional Activities Questionnaire (FAQ). Relationships between cognitive status (as indicated by MMSE score) and anxiety were explored. Anxiety was reported more commonly in patients with VaD and FTD than in patients with AD. These differences remained significant (P<0.01) in an analysis of variance (ANOVA) after adjusting for age, age at onset, educational level, and MMSE score. In AD, anxiety was inversely related to MMSE score (i.e., worse with more severe dementia), was more prevalent among patients with a younger age at onset (under age 65), and correlated with disability as measured by the FAQ score. These data suggest that anxiety is common among patients with diverse forms of dementia. In AD, anxiety is most common in those with more severe cognitive deterioration and an earlier age at onset. PMID- 12724460 TI - Item-by-item factor analysis of the Yale-Brown Obsessive Compulsive Scale Symptom Checklist. AB - Clinical subtypes of obsessive-compulsive disorder may have differing pathophysiological mechanisms and treatment outcomes. The subtypes identified by the Yale-Brown Obsessive Compulsive Scale (Y-BOCS) Symptom Checklist, clustered by clinical consensus, have never been subject to statistical validation. A factor analysis using a sample of 160 patients with OCD was conducted to determine whether factor analytically derived categories are identical to extant clinically determined categories. Our analysis revealed that the contamination subtype of the Y-BOCS Symptom Checklist contains two distinct subgroups: one relating to discomfort and the other to fear of harm. If replicated on a larger scale, the finding of new subtype groupings would have important implications for future research in obsessive-compulsive disorder. PMID- 12724461 TI - Antisocial violent offenders with attention deficit hyperactivity disorder demonstrate akathisia-like hyperactivity in three-channel actometry. AB - Actometry enables quantitative and qualitative analysis of various hyperactivity disorders. Antisocial violent offenders have demonstrated diurnal increases in motor activity that may be related to attention deficit hyperactivity disorder (ADHD) that often precedes antisocial development. Motor restlessness in ADHD has common features with neuroleptic-induced akathisia. In this study, three-channel actometry was used to compare 15 antisocial violent offenders who had a history of ADHD with 15 healthy control subjects and 10 akathisia patients. The Barnes Akathisia Rating Scale (BARS) was used for clinical evaluation of akathisia symptoms. Ankle movement indices and the ankle-waist ratio differentiated the antisocial patients from the healthy controls significantly, with no overlap, and the same parameters expectedly differentiated the akathisia patients from the healthy controls. The repetitive, rhythmic pattern of akathisia was found in 13 of the 15 antisocial patients. Nine of the antisocial patients scored 2 or 3 (mild to moderate akathisia) on the BARS. Thus, the motor hyperactivity of antisocial ADHD patients has common features with mild akathisia. This may be due to a common hypodopaminergic etiology of ADHD and akathisia. PMID- 12724462 TI - An empirical study of different diagnostic criteria for delirium among elderly medical inpatients. AB - This study compared the sensitivity and specificity of DSM-IV criteria for delirium with the sensitivity and specificity of DSM-III and ICD-10 criteria among elderly medical inpatients with or without dementia. Secondary objectives were to examine the effect of changing the definition of criterion A on sensitivity and specificity and to compare the sensitivity and specificity of different numbers of symptoms of delirium. A total of 322 elderly patients who had been admitted from the emergency department to the medical services were classified into one of four groups using DSM-III-R criteria: delirium and dementia (n = 128), delirium only (n = 40), dementia only (n = 94), and neither (n = 60). The sensitivity and specificity of DSM-IV, DSM-III, and ICD-10 criteria were determined against DSM-III-R criteria using three definitions of criterion A (clouding of consciousness only, clouding of consciousness and inattention, clouding of consciousness or inattention). When criterion A was defined as clouding of consciousness or inattention, the sensitivity and specificity of DSM IV, DSM-III, and ICD-10 criteria were 100% and 71%, 96% and 91%, and 61% and 91%, respectively. The results were similar among patients with or without dementia. The lower specificity of DSM-IV was accounted for by its inclusion of patients who did not show disorganized thinking. DSM-IV criteria for delirium are the most inclusive criteria to date for elderly medical patients with or without dementia. PMID- 12724463 TI - Longitudinal improvement in psychomotor processing speed is associated with potent combination antiretroviral therapy in HIV-1 infection. AB - This longitudinal natural history study aimed to assess the pattern and durability of neurocognitive benefits of progressively more potent combination antiretroviral therapy in HIV-1 infection. A cohort of 141 homosexual or bisexual men were assessed semiannually for CD4 count, HIV RNA viral load, medical and depressive symptoms, and a neuropsychological test battery, including psychomotor speed, verbal memory, and executive function. In a mixed-effects model, increasingly potent antiretroviral therapy was associated with improvement in tests of psychomotor processing speed. This study contributes to the growing literature documenting the longitudinal benefit provided by potent antiretroviral therapy for neuropsychological function, particularly psychomotor processing speed, in patients with HIV illness. PMID- 12724464 TI - Methamphetamine dependence is associated with neurocognitive impairment in the initial phases of abstinence. AB - This study documented the association between neurocognitive impairment and methamphetamine dependence in a sample of 27 methamphetamine-dependent individuals who achieved 5 to 14 days of continuously monitored abstinence and in 18 control subjects. Methamphetamine-dependent individuals performed significantly worse than control subjects on neurocognitive measures sensitive to attention/psychomotor speed, on measures of verbal learning and memory, and on executive systems measures sensitive to fluency. These findings are the first to demonstrate that methamphetamine dependence is associated with impairments across a range of neurocognitive domains in a sample of users whose abstinence was continuously monitored with the use of urine screening. PMID- 12724465 TI - Enhancement of declarative memory by emotional arousal and visual memory function in Alzheimer's disease. AB - The specific effects of visual and verbal memory on the ability of emotional arousal to enhance declarative memory were examined by using multiple linear regression analysis on data from a sample of 56 patients with probable Alzheimer's disease (AD). The enhancing effect of emotion on memory was evaluated by an illustrated story paradigm, and the visual and verbal memory by a standard memory test. In AD, memory enhancement by emotion was significantly correlated with visual memory but not with verbal memory, regardless of age, sex, educational attainment, and severity of dementia, suggesting a close association between memory enhancement by emotion and visual memory. PMID- 12724466 TI - Discontinuation of antidepressants in newly admitted psychotic patients. AB - Antidepressants may exacerbate manic or psychotic symptoms in vulnerable individuals. The authors discontinued antidepressants in 16 consecutive cases in which patients with manic or psychotic symptoms were otherwise judged to be on a satisfactory regimen prior to admission. Thirteen of the patients improved rapidly, which suggests a possible association between antidepressant discontinuation and clinical improvement in this patient group. PMID- 12724467 TI - Clinicopathologic case report: progressive aphasia in a 77-year-old man. PMID- 12724468 TI - Dissociative symptoms from combined treatment with sertraline and trazodone. PMID- 12724469 TI - Acute dystonia caused by low dosage of olanzapine. PMID- 12724470 TI - Favorable effect of milnacipran on depression induced by interferon-alpha. PMID- 12724473 TI - A lesion approach to neurobiology of dissociative symptoms. PMID- 12724471 TI - Counterintuitive antidepressant properties of slow rTMS over the left frontal cortex: a possible mechanism. PMID- 12724474 TI - Intermetamorphosis in a patient with Alzheimer's disease. PMID- 12724478 TI - Heart failure in aortic stenosis - improving diagnosis and treatment. PMID- 12724479 TI - Decline in invasive pneumococcal disease after the introduction of protein polysaccharide conjugate vaccine. AB - BACKGROUND: In early 2000, a protein-polysaccharide conjugate vaccine targeting seven pneumococcal serotypes was licensed in the United States for use in young children. METHODS: We examined population-based data from the Active Bacterial Core Surveillance of the Centers for Disease Control and Prevention to evaluate changes in the burden of invasive disease, defined by isolation of Streptococcus pneumoniae from a normally sterile site. Serotyping and susceptibility testing of isolates were performed. We assessed trends using data from seven geographic areas with continuous participation from 1998 through 2001 (population, 16 million). RESULTS: The rate of invasive disease dropped from an average of 24.3 cases per 100,000 persons in 1998 and 1999 to 17.3 per 100,000 in 2001. The largest decline was in children under two years of age. In this group, the rate of disease was 69 percent lower in 2001 than the base-line rate (59.0 cases per 100,000 vs. 188.0 per 100,000, P<0.001); the rate of disease caused by vaccine and vaccine-related serotypes declined by 78 percent (P<0.001) and 50 percent (P<0.001), respectively. Disease rates also fell for adults; as compared with base line, the rate of disease in 2001 was 32 percent lower for adults 20 to 39 years of age (7.6 cases per 100,000 vs. 11.2 per 100,000, P<0.001), 8 percent lower for those 40 to 64 years of age (19.7 per 100,000 vs. 21.5 per 100,000, P=0.03), and 18 percent lower for those 65 years of age or more (49.5 per 100,000 vs. 60.1 per 100,000, P<0.001). The rate of disease caused by strains that were not susceptible to penicillin was 35 percent lower in 2001 than in 1999 (4.1 cases per 100,000 vs. 6.3 per 100,000, P<0.001). CONCLUSIONS: The use of the pneumococcal conjugate vaccine is preventing disease in young children, for whom the vaccine is indicated, and may be reducing the rate of disease in adults. The vaccine provides an effective new tool for reducing disease caused by drug resistant strains. PMID- 12724480 TI - Effectiveness of pneumococcal polysaccharide vaccine in older adults. AB - BACKGROUND: Streptococcus pneumoniae is the chief cause of pneumonia in older adults, but it remains unclear whether use of the pneumococcal polysaccharide vaccine alters the overall risk of community-acquired pneumonia. In a large population of older adults, we assessed the effectiveness of the pneumococcal vaccine. METHODS: In this retrospective cohort study, 47,365 Group Health Cooperative members 65 years of age or older were assessed over a three-year period. The primary outcomes were hospitalization because of community-acquired pneumonia (validated by chart review), pneumonia in patients who were not hospitalized ("outpatient pneumonia," determined from administrative data sources), and pneumococcal bacteremia. The association between pneumococcal vaccination and the risk of each outcome was evaluated by means of multivariate Cox proportional-hazards models, with adjustment for age, sex, nursing-home residence or nonresidence, smoking status, medical conditions, and receipt or nonreceipt of influenza vaccine. RESULTS: During the study period, 1428 cohort members were hospitalized with community-acquired pneumonia, 3061 were assigned a diagnosis of outpatient pneumonia, and 61 had pneumococcal bacteremia. Receipt of the pneumococcal vaccine was associated with a significant reduction in the risk of pneumococcal bacteremia (hazard ratio, 0.56; 95 percent confidence interval, 0.33 to 0.93) but a slightly increased risk of hospitalization for pneumonia (hazard ratio, 1.14; 95 percent confidence interval, 1.02 to 1.28). Pneumococcal vaccination did not alter the risk of outpatient pneumonia (hazard ratio, 1.04; 95 percent confidence interval, 0.96 to 1.13) or of any case of community acquired pneumonia, whether or not it required hospitalization (hazard ratio, 1.07; 95 percent confidence interval, 0.99 to 1.14). CONCLUSIONS: These findings support the effectiveness of the pneumococcal polysaccharide vaccine for the prevention of bacteremia, but they suggest that alternative strategies are needed to prevent nonbacteremic pneumonia, which is a more common manifestation of pneumococcal infection in elderly persons. PMID- 12724481 TI - Nitroprusside in critically ill patients with left ventricular dysfunction and aortic stenosis. AB - BACKGROUND: Vasodilators are considered to be contraindicated in patients with severe aortic stenosis because of concern that they may precipitate life threatening hypotension. However, vasodilators such as nitroprusside may improve myocardial performance if peripheral vasoconstriction is contributing to afterload. METHODS: We determined the response to intravenous nitroprusside in 25 patients with severe aortic stenosis and left ventricular systolic dysfunction. Patients were included in the study if they had been admitted to the intensive care unit for invasive hemodynamic monitoring of heart failure and if they had a depressed ejection fraction (or=21, N=6; 19.4%). Low-complexity patients had fewer depressive and anxiety symptoms, less distress, and better clinical outcome at their 3-month follow-up than high-complexity patients. The data confirmed the ability of the instrument to detect patients at risk of complex urinary tract symptoms and to predict clinical outcome. PMID- 12724501 TI - Effect of lamotrigine on mood and cognition in patients receiving chronic exogenous corticosteroids. AB - Mood changes, cognitive deficits, and psychosis have been reported during corticosteroid therapy. However, minimal data are available on the treatment of these side effects. This pilot study examined the effect of 12 weeks of open label lamotrigine treatment (dose: mean=340 mg/day, SD=65) on mood and cognition in five patients receiving prescription corticosteroids continuously for at least 6 months before study entry. The participants showed significant improvement in cognition with lamotrigine. Two subjects who met criteria for a current major depressive episode at baseline had baseline-to-exit reductions in scores on the Hamilton Depression Rating Scale of more than 20 points. These pilot data suggest that lamotrigine may be associated with improved mood and performance on cognitive tasks in steroid-treated patients. Larger controlled trials are needed to confirm these preliminary findings. PMID- 12724502 TI - Association of depression and rheumatoid arthritis. AB - This study assessed the relative strength of the association of physical characteristics and social stresses with a diagnosis of depression in patients with rheumatoid arthritis. Depression and social difficulties were assessed in 74 patients with rheumatoid arthritis by using standardized research interviews. Rheumatoid arthritis activity, damage related to rheumatoid arthritis, and subjective functional disability were assessed with well-validated methods. Twenty-nine patients (39.2%) were depressed. Compared to nondepressed patients, depressed patients had more marked social difficulties related to rheumatoid arthritis (72.4% versus 46.7%, respectively) and more marked social difficulties independent of rheumatoid arthritis (55.2% versus 31.1%, respectively). With logistic regression, social difficulties, independent of rheumatoid arthritis, was the only variable significantly associated with depression. Demographic characteristics and rheumatoid arthritis were not associated with a diagnosis of depression. Recognition by clinicians of the importance of social stresses, independent of disease state, should lead to more appropriate and specific psychological and social treatment of depression in rheumatoid arthritis. PMID- 12724503 TI - A double-blind, placebo-controlled study of sertraline in the prevention of depression in stroke patients. AB - The authors tested the effect of sertraline in the prevention of poststroke depression. After experiencing an acute ischemic stroke, nondepressed patients (N=137) were randomly assigned to 12 months of double-blind treatment with either sertraline (N=70) or placebo (N=67). Kaplan-Meier analysis showed sertraline to have significantly superior prophylactic efficacy compared with placebo. Two definitions of clinical depression were used: total score >18 on the HAM-D(17) and score >or=9 on the HAM-D(6). Approximately 10% of the sertraline-treated group developed depression according to either definition, whereas 30% developed depression in the placebo group. On the HAM-D(6) the superiority of sertraline to placebo was demonstrated already after 6 weeks of therapy. Treatment was well tolerated; patients treated with sertraline experienced significantly fewer adverse events. PMID- 12724504 TI - Predicting panic disorder among patients with chest pain: an analysis of the literature. AB - As many as 25% of patients with chest pain who come to hospital emergency departments have panic disorder. Rates of panic disorder are even higher among those who present for outpatient evaluation of their chest pain. Unfortunately, panic disorder remains largely undiagnosed and untreated in these settings. The authors reviewed studies published between 1970 and 2001 that addressed the prevalence of panic disorder among persons who seek treatment for chest pain in an emergency department or outpatient cardiology clinic. A meta-analysis of the findings revealed five variables that appear to correlate with higher rates of panic disorder among persons who present with chest pain: 1). absence of coronary artery disease, 2). atypical quality of chest pain, 3). female sex, 4). younger age, and 5). a high level of self-reported anxiety. Further studies of these and other variables associated with panic disorder should aid in the detection of this disabling but treatable cause of chest pain. PMID- 12724505 TI - Pitfalls in assessment of decision-making capacity. AB - A total of 395 consultation-liaison psychiatrists, geriatricians, and geriatric psychologists responded to a survey that asked them to rate the frequency and importance to clinical care of 23 potential pitfalls and misunderstandings by clinicians who refer patients for assessment of decision-making capacity. Respondents also indicated which pitfalls were the most important to address in educating health care professionals. Overall, 22 of 23 pitfalls were rated as common by more than half of the respondents. Thirty-six percent of the respondents indicated that the most important pitfall to address in educating health care professionals was the tendency for health care practitioners to assume that a patient who lacks capacity for one type of medical decision also lacks capacity for all medical decisions. The results suggest that additional education is needed to improve clinicians' ability to evaluate patients' decision making capacity. PMID- 12724506 TI - Somatic symptoms for diagnosing major depression in cancer patients. AB - Diagnosing depression in cancer patients has been challenging because the diagnostic criteria include somatic symptoms frequently attributed to the cancer itself or its treatment. However, few studies have explored how to appropriately deal with individual somatic symptoms. The authors used data from 220 cancer patients with major depression to examine the intercorrelations among the DSM-IV somatic and nonsomatic symptom criteria as well as whether the presence of an individual somatic symptom could discriminate the severity of major depression. Appetite changes and a diminished ability to think were positively associated with anhedonia. Sleep disturbance and fatigue were not significantly associated with nonsomatic symptoms. These associations were consistent after adjusting for physical functioning and pain. Only patients with appetite changes showed a higher severity of depression. These results suggest that individual somatic symptoms differ in nature and that appetite-related symptoms and a diminished ability to think may be useful for diagnosing depression in cancer patients, whereas sleep disturbances and fatigue may not be as useful. PMID- 12724507 TI - Low-dose clozapine and diabetic ketoacidosis. PMID- 12724508 TI - Rapid desensitization for needle phobia. PMID- 12724509 TI - Six patterns of drug-drug interactions. AB - The literature on pharmacokinetic drug-drug interactions usually focuses on various interactions relating to the cytochrome p450 system, phase II glucuronidation, and P-glycoprotein function. However, there has been relatively little examination of how the modes or patterns that govern these interactions can be systematically characterized to better anticipate drug-drug interactions in clinical practice. This article details a schema of six core patterns of pharmacokinetic drug-drug interaction relating to processes of induction and inhibition and the action of substrates. Case examples illustrating each pattern are provided. PMID- 12724510 TI - Paradoxical reaction to fluoxetine. PMID- 12724511 TI - Itraconazole-induced delirium. PMID- 12724512 TI - Sauna and hot tub warnings. PMID- 12724513 TI - Seizures associated with venlafaxine, methylphenidate, and zolpidem. PMID- 12724514 TI - Heart transplantation and schizophrenia. PMID- 12724517 TI - Oxidative folding intermediates with nonnative disulfide bridges between adjacent cysteine residues. AB - The oxidative folding of the Amaranthus alpha-amylase inhibitor, a 32-residue cystine-knot protein with three disulfide bridges, was studied in vitro in terms of the disulfide content of the intermediate species. A nonnative vicinal disulfide bridge between cysteine residues 17 and 18 was found in three of five fully oxidized intermediates. One of these, the most abundant folding intermediate (MFI), was further analyzed by (1)H NMR spectroscopy and photochemically induced dynamic nuclear polarization, which revealed that it has a compact structure comprising slowly interconverting conformations in which some of the amino acid side chains are ordered. NMR pulsed-field gradient diffusion experiments confirmed that its hydrodynamic radius is indistinguishable from that of the native protein. Molecular modeling suggested that the eight-membered ring of the vicinal disulfide bridge in MFI may be located in a loop region very similar to those found in experimentally determined 3D structures of other proteins. We suggest that the structural constraints imposed on the folding intermediates by the nonnative disulfides, including the vicinal bridge, may play a role in directing the folding process by creating a compact fold and bringing the cysteine residues into close proximity, thus facilitating reshuffling to native disulfide bridges. PMID- 12724518 TI - Generation of neural crest-derived peripheral neurons and floor plate cells from mouse and primate embryonic stem cells. AB - To understand the range of competence of embryonic stem (ES) cell-derived neural precursors, we have examined in vitro differentiation of mouse and primate ES cells into the dorsal- (neural crest) and ventralmost (floor plate) cells of the neural axis. Stromal cell-derived inducing activity (SDIA; accumulated on PA6 stromal cells) induces cocultured ES cells to differentiate into rostral CNS tissues containing both ventral and dorsal cells. Although early exposure of SDIA treated ES cells to bone morphogenetic protein (BMP)4 suppresses neural differentiation and promotes epidermogenesis, late BMP4 exposure after the fourth day of coculture causes differentiation of neural crest cells and dorsalmost CNS cells, with autonomic system and sensory lineages induced preferentially by high and low BMP4 concentrations, respectively. In contrast, Sonic hedgehog (Shh) suppresses differentiation of neural crest lineages and promotes that of ventral CNS tissues such as motor neurons. Notably, high concentrations of Shh efficiently promote differentiation of HNF3beta(+) floor plate cells with axonal guidance activities. Thus, SDIA-treated ES cells generate naive precursors that have the competence of differentiating into the "full" dorsal-ventral range of neuroectodermal derivatives in response to patterning signals. PMID- 12724519 TI - Anthrax lethal factor represses glucocorticoid and progesterone receptor activity. AB - We report here that a bacterial toxin, anthrax lethal toxin (LeTx), at very low concentrations represses glucocorticoid receptor (GR) transactivation in a transient transfection system and the activity of an endogenous GR-regulated gene in both a cellular system and an animal model. This repression is noncompetitive and does not affect ligand binding or DNA binding, suggesting that anthrax lethal toxin (LeTx) probably exerts its effects through a cofactor(s) involved in the interaction between GR and the basal transcription machinery. LeTx-nuclear receptor repression is selective, repressing GR, progesterone receptor B (PR-B), and estrogen receptor alpha (ERalpha), but not the mineralocorticoid receptor (MR) or ERbeta. GR repression was also caused by selected p38 mitogen-activated protein (MAP) kinase inhibitors, suggesting that the LeTx action may result in part from its known inactivation of MAP kinases. Simultaneous loss of GR and other nuclear receptor activities could render an animal more susceptible to lethal or toxic effects of anthrax infection by removing the normally protective antiinflammatory effects of these hormones, similar to the increased mortality seen in animals exposed to both GR antagonists and infectious agents or bacterial products. These finding have implications for development of new treatments and prevention of the toxic effects of anthrax. PMID- 12724520 TI - Intermittent fasting dissociates beneficial effects of dietary restriction on glucose metabolism and neuronal resistance to injury from calorie intake. AB - Dietary restriction has been shown to have several health benefits including increased insulin sensitivity, stress resistance, reduced morbidity, and increased life span. The mechanism remains unknown, but the need for a long-term reduction in caloric intake to achieve these benefits has been assumed. We report that when C57BL6 mice are maintained on an intermittent fasting (alternate-day fasting) dietary-restriction regimen their overall food intake is not decreased and their body weight is maintained. Nevertheless, intermittent fasting resulted in beneficial effects that met or exceeded those of caloric restriction including reduced serum glucose and insulin levels and increased resistance of neurons in the brain to excitotoxic stress. Intermittent fasting therefore has beneficial effects on glucose regulation and neuronal resistance to injury in these mice that are independent of caloric intake. PMID- 12724521 TI - Membrane receptor trafficking: evidence of proximal and distal zones conferred by two independent endoplasmic reticulum localization signals. AB - The generic membrane trafficking signals of internal RXR and carboxyl-terminal KKXX motifs direct intracellular endoplasmic reticulum (ER) localization of the signal-bearing proteins. These signaling motifs play a critical role in partitioning proteins into designated subcellular compartments by functioning as an intracellular "zip code." In the process of determining the potential distinctions between these two otherwise functionally identical motifs, two functional zones of these signals were revealed. The KKXX signal was effective only when it was positioned closer to the membrane surface. In contrast, under identical conditions, the internal RXR signal was functional when it was positioned distally from the membrane. Different from the C-terminal KKXX signal, the internal RXR motif may be present in multiple copies. The receptor with multivalent RXR motifs displayed similar trafficking behavior to that of the same receptor with one copy of the RXR motif. The distinctive operating ranges from their anchored membrane surface provide experimental evidence for the notion that there are functional zoning layers within which membrane protein signal motifs are active. PMID- 12724522 TI - Effect of the anorectic fatty acid synthase inhibitor C75 on neuronal activity in the hypothalamus and brainstem. AB - Intraperitoneal (i.p.) injection of C75, a fatty acid synthase inhibitor, causes a rapid ( His, K8 Gly-61 --> Cys, and K18 His-127 --> Leu. The four remaining patients had mutations at one K8 and three other K18 new sites. Of the 349 blood bank control samples, only one contained the Tyr-53 --> His and one the Gly-61 --> Cys K8 mutations (P < 0.004 when comparing cirrhosis versus control groups). Two additional mutations were found in both the liver disease and blood bank groups and, hence, likely represent polymorphisms. Livers with keratin mutations had cytoplasmic filamentous deposits that were less frequent in livers without the mutations (P = 0.03). Therefore, K8K18 are likely susceptibility genes for developing cryptogenic and noncryptogenic forms of liver disease. PMID- 12724529 TI - A conserved function of YidC in the biogenesis of respiratory chain complexes. AB - The Escherichia coli inner membrane protein (IMP) YidC is involved in the membrane integration of IMPs both in concert with and independently from the Sec translocase. YidC seems to be dispensable for the assembly of Sec-dependent IMPs, and so far it has been shown to be essential only for the proper Sec-independent integration of some phage coat proteins. Here, we studied the physiological consequences of YidC depletion in an effort to understand the essential function of YidC. The loss of YidC rapidly and specifically induced the Psp stress response, which is accompanied by a reduction of the proton-motive force. This reduction is due to defects in the functional assembly of cytochrome o oxidase and the F(1)F(o) ATPase complex, which is reminiscent of the effects of mutations in the yidC homologue OXA1 in the yeast mitochondrial inner membrane. The integration of CyoA (subunit II of the cytochrome o oxidase) and F(o)c (membrane subunit of the F(1)F(o) ATPase) appeared exceptionally sensitive to depletion of YidC, suggesting that these IMPs are natural substrates of a membrane integration and assembly pathway in which YidC plays an exclusive or at least a pivotal role. PMID- 12724532 TI - Maize Genetics 2003. PMID- 12724530 TI - Unbiased quantitative proteomics of lipid rafts reveals high specificity for signaling factors. AB - Membrane lipids were once thought to be homogenously distributed in the 2D surface of a membrane, but the lipid raft theory suggests that cholesterol and sphingolipids partition away from other membrane lipids. Lipid raft theory further implicates these cholesterol-rich domains in many processes such as signaling and vesicle traffic. However, direct characterization of rafts has been difficult, because they cannot be isolated in pure form. In the first functional proteomic analysis of rafts, we use quantitative high-resolution MS to specifically detect proteins depleted from rafts by cholesterol-disrupting drugs, resulting in a set of 241 authentic lipid raft components. We detect a large proportion of signaling molecules, highly enriched versus total membranes and detergent-resistant fractions, which thus far biochemically defined rafts. Our results provide the first large-scale and unbiased evidence, to our knowledge, for the connection of rafts with signaling and place limits on the fraction of plasma membrane composed by rafts. PMID- 12724533 TI - Rha1, an Arabidopsis Rab5 homolog, plays a critical role in the vacuolar trafficking of soluble cargo proteins. AB - Rab proteins are members of the Ras superfamily of small GTP binding proteins and play important roles in various intracellular trafficking steps. We investigated the role of Rha1, an Arabidopsis Rab5 homolog, in intracellular trafficking in Arabidopsis protoplasts. In the presence of a dominant-negative mutant of Rha1, soluble vacuolar cargo proteins such as sporamin:green fluorescent protein (Spo:GFP) and Arabidopsis aleurain like protein:GFP are not delivered to the central vacuole; instead, they accumulate as a diffuse or punctate staining pattern within the cell. Spo:GFP at the punctate stains observed in the presence of hemagglutinin:Rha1[S24N] is colocalized with endogenous vacuolar sorting receptor (VSR(At-1)), which is known to localize primarily to the prevacuolar compartment, whereas Spo:GFP in the diffuse pattern is associated with tonoplasts. Furthermore, expression of Rha1[S24N] causes the secretion of a portion of the vacuolar proteins into medium. However, the inhibitory effect of Rha1[S24N] on vacuolar trafficking is relieved partially by coexpressed wild-type Rha1. Based on these results, we propose that Rha1 plays a critical role in the trafficking of soluble cargoes from the prevacuolar compartment to the central vacuole. PMID- 12724535 TI - The COP9 signalosome interacts physically with SCF COI1 and modulates jasmonate responses. AB - The COP9 signalosome (CSN) is an evolutionarily conserved, nucleus-enriched multiprotein complex. CSN plays roles in photomorphogenesis, auxin response, and floral organ formation, possibly via the regulation of ubiquitin-proteasome mediated protein degradation. COI1 encodes an F-box protein, which is a subunit of SCF(COI1) E3 ubiquitin ligase, and is required for jasmonate (JA) responses. Here, we demonstrate using coimmunoprecipitation and gel-filtration analyses that endogenous as well as epitope-tagged COI1 forms SCF(COI1) and associates directly with CSN in vivo. Like the coi1-1 mutant, CSN reduction-of-function plants exhibited a JA-insensitive root elongation phenotype and an absence of JA-induced specific gene expression. Genome expression profile analyses indicated that JA triggered genome expression is critically dependent on COI1 dosage. More importantly, most of the COI1-dependent JA-responsive genes also required CSN function, and CSN abundance was shown to be important for JA responses. Furthermore, we showed that both COI1 and CSN are essential for modulating the expression of genes in most cellular pathways responsive to JA. Thus, CSN and SCF(COI1) work together to control genome expression and promote JA responses. PMID- 12724534 TI - The COP9 signalosome interacts with SCF UFO and participates in Arabidopsis flower development. AB - The COP9 signalosome (CSN) is involved in multiple developmental processes. It interacts with SCF ubiquitin ligases and deconjugates Nedd8/Rub1 from cullins (deneddylation). CSN is highly expressed in Arabidopsis floral tissues. To investigate the role of CSN in flower development, we examined the expression pattern of CSN in developing flowers. We report here that two csn1 partially deficient Arabidopsis strains exhibit aberrant development of floral organs, decline of APETALA3 (AP3) expression, and low fertility in addition to defects in shoot and inflorescence meristems. We show that UNUSUAL FLORAL ORGANS (UFO) forms a SCF(UFO) complex, which is associated with CSN in vivo. Genetic interaction analysis indicates that CSN is necessary for the gain-of-function activity of the F-box protein UFO in AP3 activation and in floral organ transformation. Compared with the previously reported csn5 antisense and csn1 null mutants, partial deficiency of CSN1 causes a reduction in the level of CUL1 in the mutant flowers without an obvious defect in CUL1 deneddylation. We conclude that CSN is an essential regulator of Arabidopsis flower development and suggest that CSN regulates Arabidopsis flower development in part by modulating SCF(UFO)-mediated AP3 activation. PMID- 12724536 TI - Dominant-negative receptor uncovers redundancy in the Arabidopsis ERECTA Leucine rich repeat receptor-like kinase signaling pathway that regulates organ shape. AB - Arabidopsis ERECTA, a Leu-rich repeat receptor-like Ser/Thr kinase (LRR-RLK), regulates organ shape and inflorescence architecture. Here, we show that a truncated ERECTA protein that lacks the cytoplasmic kinase domain (DeltaKinase) confers dominant-negative effects when expressed under the control of the native ERECTA promoter and terminator. Transgenic plants expressing DeltaKinase displayed phenotypes, including compact inflorescence and short, blunt siliques, that are characteristic of loss-of-function erecta mutant plants. The DeltaKinase fragment migrated as a stable approximately 400-kD protein complex in the complete absence of the endogenous ERECTA protein and significantly exaggerated the growth defects of the null erecta plants. A functional LRR domain of DeltaKinase was required for dominant-negative effects. Accumulation of DeltaKinase did not interfere with another LRR-RLK signaling pathway (CLAVATA1), which operates in the same cells as ERECTA but has a distinct biological function. Both the erecta mutation and DeltaKinase expression conferred a lesser number of large, disorganized, and expanded cortex cells, which are associated with an increased level of somatic endoploidy. These findings suggest that functionally redundant RLK signaling pathways, including ERECTA, are required to fine-tune the proliferation and growth of cells in the same tissue type during Arabidopsis organogenesis. PMID- 12724537 TI - PP7 is a positive regulator of blue light signaling in Arabidopsis. AB - The cryptochrome blue light photoreceptors mediate various photomorphogenic responses in plants, including hypocotyl elongation, cotyledon expansion, and control of flowering time. The molecular mechanism of cryptochrome function in Arabidopsis is becoming increasingly clear, with recent studies showing that both CRY1 and CRY2 are localized in the nucleus and that CRY2 is regulated by blue light-dependent phosphorylation. Despite these advances, no positive cryptochrome signaling component has been identified to date. Here, we demonstrate that a novel Ser/Thr protein phosphatase (AtPP7) with high sequence similarity to the Drosophila retinal degeneration C protein phosphatase acts as an intermediate in blue light signaling. Transgenic Arabidopsis seedlings with reduced AtPP7 expression levels exhibit loss of hypocotyl growth inhibition and display limited cotyledon expansion in response to blue light irradiation. These effects are as striking as those seen in hy4 mutant seedlings, which are deficient in CRY1. We further demonstrate that AtPP7 transcript levels are not rate limiting and that AtPP7 probably acts downstream of cryptochrome in the nucleus, ensuring signal flux through the pathway. Based on our findings and recent data regarding cryptochrome action, we propose that AtPP7 acts as a positive regulator of cryptochrome signaling in Arabidopsis. PMID- 12724539 TI - Genes encoding proteins of the cation diffusion facilitator family that confer manganese tolerance. AB - The yeast Saccharomyces cerevisiae expressing a cDNA library prepared from Stylosanthes hamata was screened for enhanced Mn(2+) tolerance. From this screen, we identified four related cDNAs that encode membrane-bound proteins of the cation diffusion facilitator (CDF) family. One of these cDNAs (ShMTP1) was investigated in detail and found to confer Mn(2+) tolerance to yeast by internal sequestration rather than by efflux of Mn(2+). Expression of ShMTP1 in a range of yeast mutants suggested that it functions as a proton:Mn(2+) antiporter on the membrane of an internal organelle. Similarly, when expressed in Arabidopsis, ShMTP1 conferred Mn(2+) tolerance through internal sequestration. The ShMTP1 protein fused to green fluorescent protein was localized to the tonoplast of Arabidopsis cells but appeared to localize to the endoplasmic reticulum of yeast. We suggest that the ShMTP1 proteins are members of the CDF family involved in conferring Mn(2+) tolerance and that at least one of these proteins (ShMTP1) confers tolerance by sequestering Mn(2+) into internal organelles. PMID- 12724538 TI - The Arabidopsis SLEEPY1 gene encodes a putative F-box subunit of an SCF E3 ubiquitin ligase. AB - The Arabidopsis SLY1 (SLEEPY1) gene positively regulates gibberellin (GA) signaling. Positional cloning of SLY1 revealed that it encodes a putative F-box protein. This result suggests that SLY1 is the F-box subunit of an SCF E3 ubiquitin ligase that regulates GA responses. The DELLA domain protein RGA (repressor of ga1-3) is a repressor of GA response that appears to undergo GA stimulated protein degradation. RGA is a potential substrate of SLY1, because sly1 mutations cause a significant increase in RGA protein accumulation even after GA treatment. This result suggests SCF(SLY1)-targeted degradation of RGA through the 26S proteasome pathway. Further support for this model is provided by the observation that an rga null allele partially suppresses the sly1-10 mutant phenotype. The predicted SLY1 amino acid sequence is highly conserved among plants, indicating a key role in GA response. PMID- 12724540 TI - Conserved noncoding sequences among cultivated cereal genomes identify candidate regulatory sequence elements and patterns of promoter evolution. AB - Surveys for conserved noncoding sequences (CNS) among genes from monocot cereal species were conducted to assess the general properties of CNS in grass genomes and their correlation with known promoter regulatory elements. Initial comparisons of 11 orthologous maize-rice gene pairs found that previously defined regulatory motifs could be identified within short CNS but could not be distinguished reliably from random sequence matches. Among the different phylogenetic footprinting algorithms tested, the VISTA tool yielded the most informative alignments of noncoding sequence. VISTA was used to survey for CNS among all publicly available genomic sequences from maize, rice, wheat, barley, and sorghum, representing >300 gene comparisons. Comparisons of orthologous maize rice and maize-sorghum gene pairs identified 20 bp as a minimal length criterion for a significant CNS among grass genes, with few such CNS found to be conserved across rice, maize, sorghum, and barley. The frequency and length of cereal CNS as well as nucleotide substitution rates within CNS were consistent with the known phylogenetic distances among the species compared. The implications of these findings for the evolution of cereal gene promoter sequences and the utility of using the nearly completed rice genome sequence to predict candidate regulatory elements in other cereal genes by phylogenetic footprinting are discussed. PMID- 12724541 TI - Analysis of the Arabidopsis MADS AFFECTING FLOWERING gene family: MAF2 prevents vernalization by short periods of cold. AB - The Arabidopsis FLOWERING LOCUS C (FLC) gene is a key floral repressor in the maintenance of a vernalization response. In vernalization-sensitive genetic backgrounds, FLC levels are high, and they decline after exposure to long cold periods. Four FLC paralogs (MAF2 [MADS AFFECTING FLOWERING2] to MAF5) are arranged in a tandem array on the bottom of Arabidopsis chromosome V. We used a reverse genetics approach to analyze their functions. Loss-of-function and gain of-function studies indicate that MAF2 acts as a floral repressor. In particular, maf2 mutant plants display a pronounced vernalization response when subjected to relatively short cold periods, which are insufficient to elicit a strong flowering response in the wild type, despite producing a large reduction in FLC levels. MAF2 expression is less sensitive to vernalization than that of FLC, and its repressor activity is exerted independently or downstream of FLC transcription. Thus, MAF2 can prevent premature vernalization in response to brief cold spells. Overexpression of MAF3 or MAF4 produces alterations in flowering time that suggest that these genes also act as floral repressors and might contribute to the maintenance of a vernalization requirement. However, the final gene in the cluster, MAF5, is upregulated by vernalization. Therefore, MAF5 could play an opposite role to FLC in the vernalization response. PMID- 12724542 TI - Cloning and characterization of the WAX2 gene of Arabidopsis involved in cuticle membrane and wax production. AB - Insertional mutagenesis of Arabidopsis ecotype C24 was used to identify a novel mutant, designated wax2, that had alterations in both cuticle membrane and cuticular waxes. Arabidopsis mutants with altered cuticle membrane have not been reported previously. Compared with the wild type, the cuticle membrane of wax2 stems weighed 20.2% less, and when viewed using electron microscopy, it was 36.4% thicker, less opaque, and structurally disorganized. The total wax amount on wax2 leaves and stems was reduced by >78% and showed proportional deficiencies in the aldehydes, alkanes, secondary alcohols, and ketones, with increased acids, primary alcohols, and esters. Besides altered cuticle membranes, wax2 displayed postgenital fusion between aerial organs (especially in flower buds), reduced fertility under low humidity, increased epidermal permeability, and a reduction in stomatal index on adaxial and abaxial leaf surfaces. Thus, wax2 reveals a potential role for the cuticle as a suppressor of postgenital fusion and epidermal diffusion and as a mediator of both fertility and the development of epidermal architecture (via effects on stomatal index). The cloned WAX2 gene (verified by three independent allelic insertion mutants with identical phenotypes) codes for a predicted 632-amino acid integral membrane protein with a molecular mass of 72.3 kD and a theoretical pI of 8.78. WAX2 has six transmembrane domains, a His-rich diiron binding region at the N-terminal region, and a large soluble C-terminal domain. The N-terminal portion of WAX2 is homologous with members of the sterol desaturase family, whereas the C terminus of WAX2 is most similar to members of the short-chain dehydrogenase/reductase family. WAX2 has 32% identity to CER1, a protein required for wax production but not for cuticle membrane production. Based on these analyses, we predict that WAX2 has a metabolic function associated with both cuticle membrane and wax synthesis. These studies provide new insight into the genetics and biochemistry of plant cuticle production and elucidate new associations between the cuticle and diverse aspects of plant development. PMID- 12724543 TI - Rapid genome divergence at orthologous low molecular weight glutenin loci of the A and Am genomes of wheat. AB - To study genome evolution in wheat, we have sequenced and compared two large physical contigs of 285 and 142 kb covering orthologous low molecular weight (LMW) glutenin loci on chromosome 1AS of a diploid wheat species (Triticum monococcum subsp monococcum) and a tetraploid wheat species (Triticum turgidum subsp durum). Sequence conservation between the two species was restricted to small regions containing the orthologous LMW glutenin genes, whereas >90% of the compared sequences were not conserved. Dramatic sequence rearrangements occurred in the regions rich in repetitive elements. Dating of long terminal repeat retrotransposon insertions revealed different insertion events occurring during the last 5.5 million years in both species. These insertions are partially responsible for the lack of homology between the intergenic regions. In addition, the gene space was conserved only partially, because different predicted genes were identified on both contigs. Duplications and deletions of large fragments that might be attributable to illegitimate recombination also have contributed to the differentiation of this region in both species. The striking differences in the intergenic landscape between the A and A(m) genomes that diverged 1 to 3 million years ago provide evidence for a dynamic and rapid genome evolution in wheat species. PMID- 12724544 TI - CLAVATA1 dominant-negative alleles reveal functional overlap between multiple receptor kinases that regulate meristem and organ development. AB - The CLAVATA1 (CLV1) receptor kinase controls stem cell number and differentiation at the Arabidopsis shoot and flower meristems. Other components of the CLV1 signaling pathway include the secreted putative ligand CLV3 and the receptor-like protein CLV2. We report evidence indicating that all intermediate and strong clv1 alleles are dominant negative and likely interfere with the activity of unknown receptor kinase(s) that have functional overlap with CLV1. clv1 dominant-negative alleles show major differences from dominant-negative alleles characterized to date in animal receptor kinase signaling systems, including the lack of a dominant-negative effect of kinase domain truncation and the ability of missense mutations in the extracellular domain to act in a dominant-negative manner. We analyzed chimeric receptor kinases by fusing CLV1 and BRASSINOSTEROID INSENSITIVE1 (BRI1) coding sequences and expressing these in clv1 null backgrounds. Constructs containing the CLV1 extracellular domain and the BRI1 kinase domain were strongly dominant negative in the regulation of meristem development. Furthermore, we show that CLV1 expressed within the pedicel can partially replace the function of the ERECTA receptor kinase. We propose the presence of multiple receptors that regulate meristem development in a functionally related manner whose interactions are driven by the extracellular domains and whose activation requires the kinase domain. PMID- 12724546 TI - (E)-beta-ocimene and myrcene synthase genes of floral scent biosynthesis in snapdragon: function and expression of three terpene synthase genes of a new terpene synthase subfamily. AB - Snapdragon flowers emit two monoterpene olefins, myrcene and (E)-beta-ocimene, derived from geranyl diphosphate, in addition to a major phenylpropanoid floral scent component, methylbenzoate. Emission of these monoterpenes is regulated developmentally and follows diurnal rhythms controlled by a circadian clock. Using a functional genomics approach, we have isolated and characterized three closely related cDNAs from a snapdragon petal-specific library that encode two myrcene synthases (ama1e20 and ama0c15) and an (E)-beta-ocimene synthase (ama0a23). Although the two myrcene synthases are almost identical (98%), except for the N-terminal 13 amino acids, and are catalytically active, yielding a single monoterpene product, myrcene, only ama0c15 is expressed at a high level in flowers and contributes to floral myrcene emission. (E)-beta-Ocimene synthase is highly similar to snapdragon myrcene synthases (92% amino acid identity) and produces predominantly (E)-beta-ocimene (97% of total monoterpene olefin product) with small amounts of (Z)-beta-ocimene and myrcene. These newly isolated snapdragon monoterpene synthases, together with Arabidopsis AtTPS14 (At1g61680), define a new subfamily of the terpene synthase (TPS) family designated the Tps-g group. Members of this new Tps-g group lack the RRx(8)W motif, which is a characteristic feature of the Tps-d and Tps-b monoterpene synthases, suggesting that the reaction mechanism of Tps-g monoterpene synthase product formation does not proceed via an RR-dependent isomerization of geranyl diphosphate to 3S linalyl diphosphate, as shown previously for limonene cyclase. Analyses of tissue specific, developmental, and rhythmic expression of these monoterpene synthase genes in snapdragon flowers revealed coordinated regulation of phenylpropanoid and isoprenoid scent production. PMID- 12724545 TI - Leaf mitochondria modulate whole cell redox homeostasis, set antioxidant capacity, and determine stress resistance through altered signaling and diurnal regulation. AB - To explore the role of plant mitochondria in the regulation of cellular redox homeostasis and stress resistance, we exploited a Nicotiana sylvestris mitochondrial mutant. The cytoplasmic male-sterile mutant (CMSII) is impaired in complex I function and displays enhanced nonphosphorylating rotenone-insensitive [NAD(P)H dehydrogenases] and cyanide-insensitive (alternative oxidase) respiration. Loss of complex I function is not associated with increased oxidative stress, as shown by decreased leaf H(2)O(2) and the maintenance of glutathione and ascorbate content and redox state. However, the expression and activity of several antioxidant enzymes are modified in CMSII. In particular, diurnal patterns of alternative oxidase expression are lost, the relative importance of the different catalase isoforms is modified, and the transcripts, protein, and activity of cytosolic ascorbate peroxidase are enhanced markedly. Thus, loss of complex I function reveals effective antioxidant crosstalk and acclimation between the mitochondria and other organelles to maintain whole cell redox balance. This reorchestration of the cellular antioxidative system is associated with higher tolerance to ozone and Tobacco mosaic virus. PMID- 12724548 TI - When is a cluster of disease really a cluster? PMID- 12724549 TI - Medical emergencies at sea and injuries among Scottish fishermen. AB - BACKGROUND: It has long been known that fishing is one of the most dangerous occupations. In 2001, 33 boats were lost and 10 fishermen killed in UK waters. Despite the dangerous nature of the occupation, very little research has been conducted on fishermen's health and safety. Aims To address this gap in current knowledge, research was conducted to gain an understanding of health and lifestyle issues affecting Scottish fishermen. It was hoped that the study would identify aspects of fishermen's health that could be improved. This paper considers medical emergencies at sea and injuries among fishermen. METHODS: Data were collected using a postal questionnaire sent to the Scottish fishermen population and health diaries in a small sub-sample. RESULTS: In total, 1157 usable responses were received, giving a response rate of 57%. One-fifth of respondents had been involved in a medical emergency at sea that required them to be evacuated to shore for immediate treatment. The incidence of injuries was high, and one-third of the injuries experienced were to the back. The likelihood of evacuation for a medical emergency or experiencing an injury was increased both for certain occupations and with increasing number of boats worked on during the fisherman's career. CONCLUSION: Groups identified as being at a high risk of experiencing medical emergencies or injuries should be targeted in training initiatives or accident awareness and prevention initiatives. PMID- 12724550 TI - Ascertaining the risk of chronic obstructive pulmonary disease in relation to occupation using a case-control design. AB - BACKGROUND: Recent community surveys have reported a risk of chronic obstructive pulmonary disease (COPD) in occupations involving exposure to organic dust and gas/vapour. Aims Our aim was to confirm these results using a case-control design on 131 COPD cases (FEV(1) < 80% of predicted value, minimally reversible with bronchodilators) and 298 controls, selected from registers for patients admitted to our Institute of Occupational Medicine. METHODS: Surrogates of past exposure were occupation, exposure assigned by a job-exposure matrix, and years spent in a given occupation. The odds ratio (OR) and 95% confidence interval (CI) for COPD were calculated using logistic regression models with office workers as the reference. RESULTS: Age-smoking-adjusted ORs (and CIs) were: 15.1 (3.2-71.6) in farmers; 7.2 (1.3-41.1) in cotton workers; 6.4 (1.6-25.5) in welders; 4.7 (1.3 16.4) in painters; 12.1 (1.3-108) in foundry workers; 6.50 (1.14-37.0) in refractory brick workers; and 3.1 (1.0-9.5) in construction workers. In farmers, cotton workers, welders and painters, the adjusted ORs significantly increased (by 6-9%) for each extra year of work, while in other occupations any such increase was of borderline significance. Adjusted ORs were 3.80 (1.21-12.0), 5.83 (1.82-18.6) and 8.86 (2.29-34.3) in workers exposed to high levels of mineral dust, gas/vapour/fume and biological dust, respectively. Consistent risk estimates were obtained for farmers, textile workers, painters and welders. CONCLUSIONS: Our findings confirm previous epidemiological evidence and are supported by recent observations that cigarette smoke, cotton and organic farm dust contain the same powerful pro-inflammatory agents, and that organic dust and irritant gas induce bronchitis by triggering the same effector molecules as cigarette smoke. PMID- 12724547 TI - The GTPase ARF1p controls the sequence-specific vacuolar sorting route to the lytic vacuole. AB - We have studied the transport of soluble cargo molecules by inhibiting specific transport steps to and from the Golgi apparatus. Inhibition of export from the Golgi via coexpression of a dominant-negative GTP-restricted ARF1 mutant (Q71L) inhibits the secretion of alpha-amylase and simultaneously induces the secretion of the vacuolar protein phytepsin to the culture medium. By contrast, specific inhibition of endoplasmic reticulum export via overexpression of Sec12p or coexpression of a GTP-restricted form of Sar1p inhibits the anterograde transport of either cargo molecule in a similar manner. Increased secretion of the vacuolar protein was not observed after incubation with the drug brefeldin A or after coexpression of the GDP-restricted mutant of ARF1 (T31N). Therefore, the differential effect of inducing the secretion of one cargo molecule while inhibiting the secretion of another is dependent on the GTP hydrolysis by ARF1p and is not caused by a general inhibition of Golgi-derived COPI vesicle traffic. Moreover, we demonstrate that GTP-restricted ARF1-stimulated secretion is observed only for cargo molecules that are expected to be sorted in a BP80 dependent manner, exhibiting sequence-specific, context-independent, vacuolar sorting signals. Induced secretion of proteins carrying C-terminal vacuolar sorting signals was not observed. This finding suggests that ARF1p influences the BP80-mediated transport route to the vacuole in addition to transport steps of the default secretory pathway to the cell surface. PMID- 12724551 TI - An exploratory workplace study to investigate the perceived value of continuous low-level heatwrap therapy in manual workers. AB - BACKGROUND: Between 60 and 80% of the population in industrialized countries experience low back pain at some time in their lives. The physical demands of work, i.e. manual handling, lifting, bending, twisting, awkward postures and whole body vibration are associated with low back symptoms. This study was undertaken to investigate the benefits of a new form of continuous low-level heatwrap therapy available for the symptomatic relief of acute low back pain in the workplace. METHODS: The subjects were recruited to the study upon their visit to the occupational health doctor after being diagnosed with acute low back pain. RESULTS: The study results show that use of the heatwrap therapy significantly reduced pain intensity and impact of pain on everyday activities. The results of the study were further extrapolated to show potential cost reduction benefits of using this heatwrap therapy in the workplace. PMID- 12724552 TI - Is foundry work a risk for cardiovascular disease? A systematic review. AB - Aims Foundry work has been associated with an increased risk of adverse cardiovascular events. The objective of this review was systematically and qualitatively to review the published literature to determine whether foundry work is significantly associated with cardiac disease. METHODS: MEDLINE and Cochrane databases were systematically searched to identify relevant English language publications between 1966 and October 2002. Articles were rated as 'good', 'fair' or 'poor', using published quality review criteria. Additionally, variables suggesting causality were extracted. A qualitative summation of the literature was presented for two scenarios: all studies, or using only studies rated 'fair' and above. RESULTS: Fourteen studies were analysed. Four were found to be of 'fair' quality, the remainder 'poor'. No 'good' quality studies were found. Nine studies show increased cardiac mortality among foundry worker groups and four studies also show a decreased risk. When only 'fair' quality studies are taken into consideration, two support increased risk of cardiac disease, one supports a protective effect of foundry work on cardiac disease and one revealed both increased and decreased risk for different cardiac outcomes. CONCLUSIONS: The exploration of foundry workers' risks of cardiac events reveals conflicting findings, which can only be partly attributed to confounders. Further prospective research to establish the independent contribution of foundry work to cardiac disease is needed. PMID- 12724553 TI - Reliability and validity of instruments measuring job satisfaction--a systematic review. AB - BACKGROUND: Although job satisfaction research has been carried out for decades, no recent overview of job satisfaction instruments and their quality is available. Aim The aim of this systematic review is to select job satisfaction instruments of adequate reliability and validity for use as evaluative tools in hospital environments. METHODS: Systematic literature searches were performed in the Medline and PsycInfo databases. First, the construct of job satisfaction was operationalized by generating work factors from both theoretical studies and meta analyses or reviews of empirical studies on job satisfaction. Secondly, emphasis was placed on the internal consistency, construct validity and responsiveness of these instruments.Twenty-nine job satisfaction instruments were retrieved in total. RESULTS: Seven instruments met the defined reliability and validity criteria. Of the seven, the 'Measure of Job Satisfaction' had an adequate content validity. Only the 'Job in General Scale' provided data about 'responsiveness' to change. CONCLUSION: Few instruments have shown both high reliability and high validity, but little is known about their evaluative potential. PMID- 12724554 TI - Investigating and analysing workplace clusters of diseases: a Health & Safety Executive perspective. AB - BACKGROUND: Clusters of disease arising in workplaces cause concern among the management of the company, the workers affected and their families and friends. Chance is the most likely explanation for their occurrence, although a number of real workplace hazards have been identified through their observation and investigation. Employers have a duty to investigate such occurrences in order to assess whether some unknown or unidentified hazard is at work and to take the appropriate action. Several papers have been published over the last 15 years or so that set out a method for investigating workplace clusters of disease. Aims This paper presents the steps in the approach taken by the Health & Safety Executive in Great Britain. METHOD: An initial step identifies the relevant stakeholders at the outset, in order to maintain a realistic expectation of what the investigation can hope to achieve and to open a dialogue. The main steps in the assessment are: (1) identifying cases; (2) determining the other parameters of the investigation; (3) statistically assessing the cluster; (4) examining potential exposures and assessing their biological plausibility; and (5) determining the overall significance of the cluster. The approach is illustrated throughout by examples. PMID- 12724555 TI - The rise and fall in incidence of malignant mesothelioma from a British Naval Dockyard, 1979-1999. AB - BACKGROUND: The incidence of malignant mesothelioma in Britain is predicted to rise over the next 15-25 years because of past failure to protect the workforce against inhalation of asbestos. In British Naval dockyards, alternative insulation materials and respiratory protection were introduced from the mid 1960s. Aims This study was carried out to investigate the effects of these control measures on mesothelioma deaths in dockyard workers. METHODS: Cases of mesothelioma of the pleura and peritoneum between 1979 and 1999 in workers from the Devonport Naval Dockyard, south-west England, were sought from coroners' and medico-legal records. RESULTS: Three hundred and one cases were identified, 7% peritoneal. The peak incidence occurred in 1991 with 25 cases per annum (quadratic model fit R(2) = 74.2%, P < 0.001) and we predict that by 2003 the incidence will fall to fewer than five cases per annum. The mean time between first exposure and presentation was 48.5 years [95% confidence interval (CI) = 47.3-49.8], but this was significantly shorter in the more heavily exposed trades, when compared with the less heavily exposed (42 years, 95% CI = 39.0 45.0, versus 49.5 years, 95% CI = 48.2-50.9). Those with higher exposure were also at significantly greater risk of peritoneal disease (P < 0.023, Fisher's exact test). CONCLUSION: The reduction in incidence of mesothelioma is greater than can be accounted for by reduction in numbers of dockyard workers over the last 50 years. Changes in insulation materials and improved industrial hygiene measures introduced into the Devonport Dockyard from the mid-1960s have resulted in an earlier decline in the incidence of malignant mesothelioma than that predicted for the British workforce as a whole. PMID- 12724556 TI - Demographic and lifestyle predictors of body mass index among offshore oil industry workers: cross-sectional and longitudinal findings. AB - BACKGROUND: Significant overweight among offshore workers on North Sea oil and gas installations has been linked to high calorie intake, lack of active leisure time pursuits, and environmental factors conducive to weight gain. However, the prevalence of overweight among offshore workers has not been examined in recent data, and no longitudinal studies of body mass index (BMI) in this occupational group have been reported. Aims The present study sought to examine BMI levels in a sample of UK offshore personnel, and to evaluate demographic factors, smoking and work-related physical activity as predictors of BMI, and 5 year change in BMI. METHODS: Survey data (including age, education, marital status, work-related physical activity and height/weight) were collected in 1995 from male workers on 17 North Sea installations (n = 1581, 83% response rate); follow-up data were obtained in 2000 (n = 354, 34.9% of the potential sample). RESULTS: Overall mean BMI was 25.6 (2.8) kg/m(2): rates of obesity (BMI > 30) and overweight (BMI = 25 30) were 7.5 and 47.3%, respectively. Mean age was 38.7 (8.9) years; linear and quadratic age terms predicted BMI. Age-adjusted BMI values were very similar to those reported from other offshore studies over the past 15 years. Age, marital status, education, smoking and physical activity significantly predicted baseline BMI, but only age (and some interactive effects) predicted 5 year BMI change. CONCLUSIONS: The present age-adjusted BMI values were closely similar to those found offshore in the mid-1980s, but also to recent national data; thus, North Sea personnel do not appear to reflect current population trends towards increased BMI levels. This result accords with the emphasis now given to health promotion (particularly dietary change) on offshore installations; the present findings also highlight the need to focus these initiatives on workers with sedentary jobs and/or low education. PMID- 12724557 TI - Measurement of beryllium in lung tissue of a chronic beryllium disease case and cases with sarcoidosis. AB - BACKGROUND: The clinical features of chronic beryllium disease (CBD) are similar to many other chronic lung diseases. In particular, it may be difficult to distinguish it from pulmonary sarcoidosis since the two conditions may be very alike in clinical, pathological and radiological features. Aim To determine if the amount of beryllium found in the lungs could be used to differentiate CBD from sarcoidosis and controls. METHODS: Analyses for beryllium in the autopsied lung tissues of 29 cases and controls were carried out. The cases included one CBD, three confirmed sarcoidosis and 25 controls. Blocks of formalin-fixed tissues were analysed by an atomic absorption spectrophotometer equipped with a graphite furnace. A method for analysis of beryllium in air was modified to permit tissue analysis. RESULTS: The CBD case had a much higher average beryllium level, but some individual results were similar to controls and patients with sarcoidosis. CONCLUSION: The CBD case had beryllium levels within the range of values reported in the literature. The differentiation between CBD and sarcoidosis could not be made with reasonable assurance based only on the analytic result. Occupational history is very important in making a diagnosis of CBD, along with the analysis of tissues. Tissue analysis helped confirm the diagnosis of compensatable CBD in this particular case. PMID- 12724558 TI - The connection between age, job control and sickness absences among Finnish food workers. AB - METHODS: The relationship between sickness absence and job control among industrial food workers of different ages (n = 114) was studied. RESULTS: The number of absence spells, particularly short absence spells, was higher among younger workers, as expected. Using multivariate analysis, low job control was associated with an increased number of all absence spells (P < 0.02). CONCLUSIONS: Job control may be an important factor in determining sickness absence among industrial workers. PMID- 12724559 TI - Aesthesioneuroblastoma in a woodworker. AB - A case is described of aesthesioneuroblastoma in a woodworker who had been exposed to wood dust for 25 years, without any individual or environmental protection. The case described supports the contention that occupational exposure to wood dust may have caused the neoplasm. PMID- 12724560 TI - Difficult to access does not mean disadvantaged. PMID- 12724561 TI - Monitor. PMID- 12724562 TI - In situ zymography: a molecular pathology technique to localize endogenous protease activity in tissue sections. AB - Proteases play important roles in modulating a wide range of cellular functions, in the regulation of biologic processes, and in the pathogenesis of various diseases. Several molecular techniques are available to identify and characterize proteases in cells and tissues. Most of these techniques do not provide information on the activity of proteases in tissues. In situ zymography (ISZ) is a relatively low-cost technique that uses specific protease substrates to detect and localize specific protease activities in tissue sections. Used in combination with other techniques, ISZ provides data that further our understanding of the role of specific proteases in various pathologic and physiologic conditions. This review describes the general principle of ISZ and highlights the past and future applications of this technique in molecular pathology. PMID- 12724563 TI - Clinical and pathologic relevance of p53 index in canine osseous tumors. AB - The clinicopathologic value of the immunohistochemical (IHC) expression of p53 protein was evaluated in 167 canine osseous tumors. p53 staining frequency and intensity in tumor cells was expressed as a p53 index. p53 index was significantly higher in osteosarcomas than in other sarcomas, chondrosarcoma, multilobular tumor of bone, and tumors initially misdiagnosed as osteosarcomas as well as in appendicular versus axial and in distal versus proximal osteosarcomas. A strong correlation is demonstrated between the p53 index and a range of clinicopathologic parameters in osteosarcoma, including the tumor site, histologic grade and score, mitotic index, degree of tumor necrosis, and pleomorphism. Chondroblastic osteosarcomas had significantly higher and telangiectatic osteosarcomas significantly lower p53 index than did osteosarcomas belonging to other histopathologic subtypes, a fact that tends to reinforce the perception of these osteosarcomas as distinct clinicopathologic entities. Entire males had higher p53 index than did neutered males. p53 index was higher in Rottweilers than in Great Danes and Terriers, confirming breed susceptibilities to osteosarcoma. p53 index showed no association with age, primary or secondary site status, or the presence of metastases or other tumor types. Biopsy samples had a higher p53 index than did postmortem samples, either because of differences in sample processing or the possibility that p53 overexpression is more evident at the earlier stages of osteosarcoma pathogenesis, presumably represented by the biopsy material. IHC examination for p53 and the derived index has the potential to be used as an additional diagnostic tool and prognostic indicator for osseous tumors. PMID- 12724564 TI - Histopathologic and immunophenotypic characterization of extramedullary plasmacytomas in nine cats. AB - To contribute to the existing body of knowledge in the literature on the apparently rare extramedullary plasmacytoma in cats, lymphoid tumors with plasmacytic cellular morphology taken from nine cats were examined. The paraffin embedded material was investigated by standard hematoxylin and eosin, and special staining techniques (Giemsa, Congo-red, and periodic acid-Schiff reaction). The tumors also were examined immunohistochemically for the presence of immunoglobulin G, immunoglobulin A, immunoglobulin M, immunoglobulin light chains (lambda, kappa), various amyloid proteins, and FeLV-antigen (p27 protein). An immunoglobulin-producing tumor of plasmacellular origin (extramedullary plasmacytoma [EMP]) could be diagnosed in all cases on the basis of immunohistochemical light-chain expression. All but one of the neoplasms occurred in the skin of older, predominantly male cats. As in humans and dogs, the following types could be identified according to their morphologic features: mature type (two), cleaved type (two), asynchronous type (four), and polymorphous type (one). The tumor tissue of three cats revealed amyloid deposits, which were immunohistochemically diagnosed as ALlambda-amyloid in all three cases. PMID- 12724565 TI - A histopathologic, immunohistochemical, and ultrastructural study of the intestine in pigs inoculated with classical swine fever virus. AB - The aim of this study was to report on the lesions occurring in the intestine during experimental classical swine fever (CSF) and to clarify the nature of infected cells and the distribution of viral antigen. Thirty-two pigs were inoculated with the virulent CSF virus (CSFV) isolate Alfort 187 and slaughtered from 2 to 15 postinoculation days; four animals of similar background served as a control group. Immunohistochemistry, electron microscopy, and the transferase mediated deoxyuridine triphosphate nick-end labeling method were used to detect viral antigens and apoptosis. The results showed progressive lymphoid depletion and mucosal necrosis. The lymphoid depletion could have been caused by apoptosis of lymphocytes but could not be directly attributed to the effect of CSFV on these cells. Vascular changes, pathogenic bacteria, and viral infection of epithelial cells were ruled out as causes of necrotic lesions. However, large virally infected monocytes-macrophages with ultrastructural changes indicative of activation were observed in the intestine. This suggests that monocytes macrophages play an important role in the pathogenesis of intestinal lesions. An understanding of the function of these cells will require additional study. PMID- 12724566 TI - Experimental infection of equine herpesvirus 9 in dogs. AB - Equine herpesvirus 9 (EHV-9), a new neurotropic equine herpesvirus, was inoculated intranasally at 107 plaque-forming units in five dogs to assess its pathogenicity. Dogs showed weight loss, pyrexia, anorexia, and neurologic signs on the fourth day. The EHV-9 virus was recovered from the examined brains. Histologically, dogs had a fulminant nonsuppurative encephalitis characterized by severe neuronal degeneration and loss, with intranuclear inclusions, slight glial reactions, perivascular cuffing, and multifocal hemorrhage. The olfactory bulb and the frontal and temporal lobes were predominantly affected. Immunohistochemistry revealed reactivity for EHV-9 antigen in neurons. All dogs had mild bronchopneumonia and various degrees of lymphoid necrosis. These findings indicate that dogs are fully susceptible to EHV-9 and that EHV-9 can cause fulminant encephalitis with high mortality in dogs, as in gazelles and goats. PMID- 12724567 TI - Morphometry of canine cutaneous mast cell tumors. AB - Twenty-four canine cutaneous nodules, diagnosed as mast cell tumors by fine needle aspiration biopsy and confirmed by histopathologic analysis by staining with hematoxylin and eosin (HE) and toluidine blue, were analyzed by computerized nuclear morphometry on panoptic- and HE-stained cytopathology slides. Two hundred nuclei per lesion were examined. The morphometric parameters investigated were nuclear area, mean diameter, perimeter, regularity factor, and ellipticity factor. Lesions were graded as I (well differentiated), II (intermediate differentiation), or III (poorly differentiated) according to the following morphologic features: invasiveness, cellularity and cellular morphology, mitotic index, and stromal reaction. Nuclear morphometric results were then compared with histopathologic grades. Values of nuclear area, mean diameter, and perimeter increased with increase in histopathologic grade, but statistical analysis revealed significant differences only between grades II and III and between grades I and III when HE was used (P < 0.01) and between grades I and III with panoptic stain (P < 0.05). The ellipticity factor and regularity factor did not reveal significant differences between histopathologic grades. The results indicate that nuclear morphometric analysis, in combination with the rapid and inexpensive cytopathology technique, can help in mast cell tumor grading, thus contributing to the establishment of a more precise prognosis and treatment. PMID- 12724568 TI - Evidence of nitric oxide synthase 2 activity in swine naturally infected with Actinobacillus pleuropneumoniae. AB - Evidence of nitric oxide synthase (NOS) 2 activity was determined by formation of nitrotyrosine (a reaction product of peroxynitrite) and by activation of poly(ADP ribose) synthetase (PARS) in NOS2-expressed pleuropneumonic lungs from 20 pigs naturally infected with Actinobacillus pleuropneumoniae using immunohistochemistry. Intense immunostaining for nitrotyrosine residue was seen within the lung lesions from A. pleuropneumoniae-infected pigs, but it was minimal in the unaffected parts of the lung from A. pleuropneumoniae-infected pigs and in the normal lung from control pigs. Staining was especially strong in neutrophils and macrophages in the periphery of the lesions and within the alveolar spaces. There was close cell-to-cell correlation when serial sections were examined by immunohistochemistry for NOS2 and nitrotyrosine in each of the 20 lung samples. Expression of PARS was always present within inflammatory lesions but was minimal in the unaffected lung of A. pleuropneumoniae-infected pigs. Macrophages in alveolar spaces frequently exhibited strong staining for PARS. Colocalization of nitrotyrosine and PARS antigen was especially prominent in macrophages in the periphery of lesions. NOS2 expression in pleuropneumonic areas associated with protein nitrosation and PARS suggests that NOS2 is functionally active during infections caused by A. pleuropneumoniae. PMID- 12724569 TI - Abscess-forming inflammatory granulation tissue with Gram-positive cocci and prominent eosinophil infiltration in cats: possible infection of methicillin resistant Staphylococcus. AB - We occasionally encounter feline cervical or mesenteric lesions diagnosed histopathologically as abscess or inflammatory granulation tissue with eosinophil infiltration. Gram-positive cocci accompany the lesions. In the present study, such lesions obtained from 27 cats were examined to evaluate the histopathologic features and the nature of the causative bacteria. The average age was 7.3 +/- 3.5 years. No sex predilection was observed. Most frequent locations of the lesions included the abdominal cavity with/without mesenteric lymph nodes (11/27, 41%) and subcutaneous tissue or lymph nodes of the neck (9/27, 33%). Common clinical presentation was a localized mass. Grossly, the lesions contained abscesses in the center and were surrounded by fibrous tissue. Microscopically, the necrotic zone contained bacterial colonies. Large numbers of eosinophils and macrophages infiltrated the area surrounding the necrotic tissue. The surrounding connective fiber-rich granulation tissue demarcated the eosinophilic abscess. The bacteria were Gram-positive cocci in 23 of the 27 cats and were positive for anti staphylococcus antiserum in 19 of the 23 cats. In 15 out of 17 lesions, the colonies expressed immunoreactivity to penicillin-binding protein 2', which is a drug-resistance gene product of methicillin-resistant Staphylococcus (MRS) species. These findings suggest strongly that MRS causes this type of infectious lesion. PMID- 12724570 TI - Histology and immunohistochemistry of seven ferret vaccination-site fibrosarcomas. AB - The anatomical location, histology, and immunohistochemistry of 10 ferret dermal and subcutaneous fibrosarcomas were examined. Seven of the 10 tumors were from locations used for vaccination. All fibrosarcomas contained spindle-shaped cells surrounded by variable quantities of connective tissue stroma. However, vaccination-site fibrosarcomas (VSFs) subjectively contained a higher degree of cellular pleomorphism. Multinucleated cells were present in three of seven VSFs but not in any of the nonvaccination-site fibrosarcomas (NVSFs). Large histiocytic cells, interpreted as macrophages, containing intracytoplasmic basophilic granular material were observed in two VSFs but not in any of the NVSFs. Five VSFs contained peripheral lymphoplasmacytic aggregates. Immunohistochemically, three VSFs stained with anti-smooth muscle actin antibodies and one stained with antibodies against desmin. No expression of muscle cytoskeletal filaments was observed in any NVSF. Filaments interpreted as actin were visible in both the VSFs examined ultrastructurally. One of the VSFs examined ultrastructurally contained intracytoplasmic crystalline material. The preferential development of subcutaneous fibrosarcomas in vaccination sites suggests that, as in cats, vaccination may promote local sarcoma development in ferrets. Additionally, some of the histologic, immunohistochemical, and ultrastructural features of these tumors are similar to those reported for feline vaccine-associated sarcomas. To the authors' knowledge, vaccination has not previously been reported to be oncogenic in any species other than cats. PMID- 12724571 TI - Adenoviral gizzard erosion in commercial broiler chickens. AB - Pathologic and immunohistochemical changes caused by group I of the fowl adenovirus (FAV) serotype-1 99ZH strain, isolated from broiler chickens exhibiting gizzard erosion, were investigated in commercial broiler chickens. One hundred twenty-two chickens were inoculated with the strain by both oral and ocular routes at 1, 3, or 5 weeks of age and euthanatized for necropsy within 4 18 days of inoculation. Focal gizzard erosions were observed in the inoculated chickens of each age group. A histologically degenerative koilin layer, necrotic mucosa, intranuclear inclusion bodies in the glandular epithelial cells, inflammatory cell infiltrations in the lamina propria, submucosa, and a muscle layer were seen in the gizzards. Immunohistochemical staining showed evidence of FAV antigens in the intranuclear inclusion bodies. These findings were recognized regardless of their maternal antibody levels for FAV serotype-1. Gizzard lesions appeared later in the lower-dose-inoculated chickens than in the higher-dose inoculated chickens. Numerous CD3-positive cells and IgY-positive plasma cells were seen in the gizzard lesions. In 5-week-old chickens the heterophil infiltrations in the lesions were milder than in younger chickens. Intranuclear inclusion bodies also were observed in the epithelial cells of the ileum or cecal tonsils of some chickens. Thus, this study shows that FAV-99ZH causes adenoviral gizzard erosion in broiler chickens without hepatic or pancreatic lesions and that cell infiltration is more severe than in dietary gizzard erosions. PMID- 12724572 TI - Pathology of human influenza A (H5N1) virus infection in cynomolgus macaques (Macaca fascicularis). AB - Infection with influenza A (H5N1) virus, which has not been associated with respiratory disease in humans previously, caused clinical signs of acute respiratory distress syndrome and multiple-organ dysfunction syndrome with high mortality in humans in Hong Kong in 1997. To study the pathogenesis of this disease, we infected four cynomolgus monkeys (Macaca fascicularis) with 2.5 x 104 median tissue culture infectious dose (TCID50) of influenza virus A/Hong Kong/156/97 (H5N1) and euthanatized them 4 or 7 days after infection. The main lesion was a necrotizing broncho-interstitial pneumonia (4/4) similar to those found in primary influenza virus pneumonia in humans, with desquamation of respiratory epithelium (4/4), intra-alveolar hemorrhage (4/4), hyaline membrane formation (2/4), and infiltration with neutrophils and macrophages (4/4). Lesions in other organs consisted of a suppurative tonsillitis (2/4) and necrosis in lymphoid organs (1/4), kidney (1/4), and liver (1/4). By immunohistochemistry, influenza virus antigen was limited to pulmonary tissue (4/4) and tonsils (2/4). Based on these results, we suggest that the cynomolgus monkey is a suitable animal model for studying the pathogenesis of human H5N1 virus infection and that multiple-organ dysfunction syndrome in this disease may be caused by diffuse alveolar damage from virus replication in the lungs alone. PMID- 12724573 TI - Symmetric focal degeneration in the cerebellar and vestibular nuclei in swine caused by ingestion of Aeschynomene indica seeds. AB - A vestibulocerebellar disorder was observed in 2- to 4-month-old swine after consumption of broken rice contaminated with 13% Aeschynomene indica seeds. Affected animals recovered in 2-14 days after removal of the contaminated food. To reproduce the syndrome, 10 pigs were fed 13% A. indica seeds in commercial pig ration or rice. They showed clinical signs similar to those observed in the spontaneous cases, 1 hour to 6 days after ingestion. Three pigs recovered after the withdrawal of the contaminated food. The others were killed. Two pigs that were fed for 25 days with food containing 3% and 6% seeds and two control pigs did not show clinical signs. Histologic lesions were characterized by symmetric focal degeneration in the cerebellar and vestibular nuclei. The initial lesion, observed 24 hours after the first ingestion, was characterized by vacuolation of the neuropil. Subsequently, there was progressive loss of parenchyma, vascular reaction, a few small spheroids, astrocytosis, and accumulation of gitter cells. The brains from four pigs were perfused with a buffered solution of glutaraldehyde-paraformaldehyde 12-48 hours after they started to ingest the seeds. No ultrastructural lesions were observed in the cerebellar nuclei after 12 hours of seed consumption. At 24 hours, endothelial cells and pericytes were separated by empty spaces from astrocyte foot processes, suggesting perivascular edema. The astrocytes were enlarged, consistent with intracellular fluid accumulation. These results suggest that A. indica seeds are toxic, causing functional derangement of the vestibulocerebellar system, followed by alterations in the microvasculature in the cerebellar and vestibular nuclei. PMID- 12724574 TI - Narthecium ossifragum (L.) huds. causes kidney damage in goats: morphologic and functional effects. AB - We studied the effects of Narthecium ossifragum on goat kidneys. Twenty-five Norwegian dairy goats, 5 weeks to 4 months of age, were orally dosed with an aqueous extract from N. ossifragum. In experiment 1, we studied microscopic and functional changes in 12 animals that were euthanatized 2, 3, 4, 5, and 6 days after treatment. In experiment 2, we included ultrastructural studies on serial renal biopsies and urine analysis from five extract-treated animals and two controls. In addition, urine samples were collected from four dosed and two control goats. Ultrasonography revealed perirenal and retroperitoneal fluids. Microscopic changes were observed after 6 hours. The findings, most obvious in the inner cortex and the outer medulla, consisted of cytoplasmic vacuolization, interstitial edema, and focal necrosis of tubular epithelial cells. Ultrastructurally, the tubules had loss of microvilli, irregular cytoplasmic vacuolization, mitochondrial swelling with loss of cristae, and irregular but continuous basement membranes even with necrosis. In the glomeruli, there were occasional endothelial damage and shortening and swelling of the foot processes. Peritubular capillaries had breaks in the vessel walls and irregular endothelial cell edema, and the interstitium had marked edema. The functional lesions included elevated serum urea, creatinine, and magnesium concentrations, a slight decrease in serum calcium concentration, elevated urine protein and urine protein creatinine ratio, and increased activities of urine alkaline phosphatase and gamma glutamyl transferase. Our findings indicate a fast-acting toxic principle inducing damage by both direct toxic and secondary ischemic effects. PMID- 12724575 TI - Immunohistochemical detection of apolipoprotein A-I and B-100 in canine atherosclerotic lesions. AB - We attempt to determine and compare the localization of apolipoproteins (apo) apoA-I and B-100 in atherosclerotic lesions of canine aortas, coronary arteries, and the peripheral arteries, using immunohistochemical techniques. Histopathologically, atherosclerotic lesions were characterized by deposition of lipids and infiltration of lipid-laden foamy cells in the tunica intima and tunica media, sometimes forming fibrofatty plaques containing abundant sudanophilic and mineralized material. Canine apoA (CapoA)-I and canine apoB (CapoB)-100 immunopositive signals were simultaneously observed in mild and severe atherosclerotic lesions of the aorta, coronary arteries, splenic arteries, and renal arteries in the double-immunolabeled sections. Both CapoA-I and CapoB 100 positive signals were seen in the cytoplasm of endothelial cells, smooth muscle cells, and macrophages. The subendothelial space and extracellular matrix in the tunica intima and media were also positive. Neither CapoA-I nor CapoB-100 positive signals were seen in normal arteries. These findings closely resemble those of the localization of apoA-I and apoB-100 in human atherosclerotic lesions. PMID- 12724577 TI - Hemochromatosis secondary to repeated blood transfusions in a dog. AB - Hemochromatosis was presumptively diagnosed using cytologic examination of liver tissue from an aged male Miniature Schnauzer. The dog was presented after receiving whole blood transfusion every 6-8 weeks for 3 years to treat pure red cell aplasia. The cytologic specimen contained clusters of hepatocytes with abundant intracytoplasmic gold-yellow pigment granules and clumps of extracellular, green-black, globular pigment, both interpreted to be hemosiderin. Histologic sections of liver revealed hepatocellular degeneration with bridging portal fibrosis, lobular atrophy, biliary hyperplasia, and diffuse, severe hemosiderin accumulation. Serum iron and ferritin levels, and dry-weight iron concentrations of liver, heart, and kidneys were markedly increased. Hemosiderin accumulation was confirmed in hepatocytes of cytologic and histologic specimens using Perl's Prussian blue staining. This report is the first description of transfusional hemochromatosis in a dog and is the first to describe its cytologic appearance in a veterinary patient. PMID- 12724576 TI - Squamous cell carcinoma of the anal sac in five dogs. AB - Tumors of the perianal area of dogs are common and include multiple tumor types. Whereas perianal adenomas occur often, adenocarcinomas of the apocrine glands of the anal sac occur less frequently. A review of the literature revealed no reports of squamous cell carcinomas arising from the epithelial lining of the anal sac. Squamous cell carcinomas originating from the lining of the anal sac were diagnosed in five dogs. Microscopically, the tumors consisted of variably sized invasive nests and cords of epithelial cells displaying squamous differentiation. Four of the five dogs were euthanatized because of problems associated with local infiltration by the tumors. In the fifth dog, there was no evidence of tumor 7 months after surgical removal, but further follow up was not available. PMID- 12724579 TI - Gastric amebiasis due to Entamoeba histolytica in a Dama wallaby (Macropus eugenii). AB - A 1.5-year-old captive female Dama wallaby (Macropus eugenii) died after a 3 month period of progressive weight loss, anorexia, bloat, and diarrhea. Histopathologic examination revealed numerous Entamoeba histolytica trophozoites within the gastric mucosa and, less frequently, gastric submucosa and submucosal vessels. Immunofluorescent antibody testing confirmed the identity of the trophozoites as E. histolytica. The trophozoites were associated with mild glandular epithelial necrosis, mucosal erosions, and lymphoplasmacytic inflammation. E. histolytica most commonly causes necrotizing and ulcerative colitis in humans and captive nonhuman primates, and it causes necrotizing and ulcerative gastritis in nonhuman primates with sacculated stomachs adapted for leaf fermentation. Rare cases of gastric amebiasis also have been been reported in captive macropods, which also have complex sacculated stomachs. To our knowledge, this is the first report confirming E. histolytica as the cause of gastric amebiasis in a wallaby. The zoonotic potential of this infection in macropods is uncertain. PMID- 12724578 TI - Septic peritonitis due to colonic perforation associated with aberrant migration of a Gasterophilus intestinalis larva in a horse. AB - An adult quarter horse mare was presented with acute colic and fever. Physical examination and abdominocentesis showed septic peritonitis, and the mare was euthanatized. Necropsy and histopathologic examination revealed a focal partial perforation of the right ventral colon, which contained a single bot fly larva, identified as a third-instar larva of Gasterophilus intestinalis. This larva was embedded deep within the muscularis and the submucosa. Although bot fly larvae are known to attach to aberrant sites within the digestive tract, this is the first known report of deep penetration of the colon by a gasterophilus larva, with the resulting leakage of intestinal content leading to septic peritonitis. PMID- 12724580 TI - Diffuse coronary disease and atherothrombosis: a rationale for long-term therapy to prevent recurrent ischemic events. PMID- 12724581 TI - Antiplatelet therapy: aspirin. PMID- 12724582 TI - Aspirin and clopidogrel in patients with ACS undergoing PCI: CURE and PCI-CURE. PMID- 12724583 TI - Improving acute coronary syndrome care: the ACC/AHA guidelines and critical pathways. PMID- 12724584 TI - Brachytherapy and drug-eluting stents. PMID- 12724585 TI - Case presentations and discussions. PMID- 12724589 TI - Gait analysis alters decision-making in cerebral palsy. AB - This study was designed to assess the impact of gait analysis on the treatment of patients with cerebral palsy. One hundred two ambulant patients with cerebral palsy were assessed clinically and with gait analysis. Separate treatment proposals for each patient were recorded after clinical examination and after gait analysis. The results of the two methods of assessment were compared. After clinical assessment, 71 of the 102 patients evaluated were recommended for a surgical procedure and 31 for nonoperative treatment. After gait analysis, the indications for treatment were confirmed in 91 cases (89%). Clinical assessment by the same orthopedic surgeon was in close agreement with gait analysis in identifying an indication for surgery. There was less agreement in the type or level of operation recommended. Gait analysis altered the decision in 106 of 267 operations (40%). There was good agreement for bone surgery, suggesting that clinical evaluation of torsional problems was fairly reliable. The poorer agreement seen for soft tissue operations probably reflects the difficulties in assessing tone-related problems in these patients clinically. This study confirms the value of gait analysis for decision-making in cerebral palsy. PMID- 12724590 TI - Edinburgh visual gait score for use in cerebral palsy. AB - Complex gait analysis systems are not generally available worldwide, and no simple system of assessing gait by observation has been validated specifically for use in patients with cerebral palsy. The authors have developed a visual gait analysis score for use in cerebral palsy. Videotaped sequences of patients were recorded before and after surgery as part of a three-dimensional gait study using a Vicon (Oxford, U.K.) gait analysis system. The score demonstrated good intraobserver and interobserver reliability. The numeric values of the score elements correlated well with the measurements obtained from instrumented gait analysis for the same patients, and the score was able to detect postoperative change. PMID- 12724586 TI - Interobserver variability of gait analysis in patients with cerebral palsy. AB - In this study 11 ambulatory patients (mean 10.8 years) with spastic cerebral palsy were each evaluated with instrumented gait analysis at four different centers. After review of the data, each medical director chose from a list of treatment options. The average variability in static range of motion from physical examination ranged from 25 degrees to 50 degrees. Hip and knee sagittal motion had the best relative variability of 20 degrees to 24%. Via gait analysis, the average variability in sagittal, coronal, and transverse plane kinematic motions averaged 12, degrees 7 degrees, and 20 degrees, respectively. Increased variability was noted in transverse (worst) to coronal and finally sagittal (best) plane motion. Only two mildly affected patients had similar, but not exact, treatment recommendations. The authors conclude that substantial variations in raw data exist when the same cerebral palsy patient is evaluated at different gait centers. These data do not yield the same treatment recommendations in the majority of patients. PMID- 12724591 TI - Walking speed in children and young adults with neuromuscular disease: comparison between two assessment methods. AB - Self-selected walking speed is being increasingly used as a primary outcome measure in the management of neuromuscular disease. It would be useful if the speed recorded in the gait laboratory represented the child's walking speed in the community. This study investigated the difference in self-selected walking speeds between a 10-meter walk, as measured during instrumented gait analysis, and a 10-minute walk. The authors found that self-selected walking speed during the 10-minute walk was slower than the self-selected walking speed recorded during the 10-meter walk. The former may be more representative of walking speed in the community setting. Walking speed measured during walks of 10 minutes or more should become an integral part of gait laboratory evaluation. PMID- 12724592 TI - Effects of surgical lengthening of the hamstrings without a concomitant distal rectus femoris transfer in ambulant patients with cerebral palsy. AB - Eighteen ambulant patients (32 legs) who had undergone fractional lengthening of the medial and lateral hamstrings without rectus femoris transfer underwent pre- and postoperative gait analysis. A significant increase in the amount of knee extension and a decrease in the amount of peak knee flexion in swing were observed. This decrease in knee flexion signified a change towards more normal speed-related values. Dorsiflexion at initial contact decreased significantly for patients who did not undergo a gastrocnemius lengthening (n = 24). Absolute cadence was significantly lower after surgery, but the change in dimensionless cadence was not significantly different. This difference in the outcome between dimensionless and absolute stride parameters can be attributed to the increase in body height after surgery. The clinical significance of these findings is that it is important to recognize that postoperative effects of surgery on gait in children may, in part, be explained by changes in height and not surgery alone. PMID- 12724593 TI - Spastic hemiplegic cerebral palsy and the femoral derotation osteotomy: effect at the pelvis and hip in the transverse plane during gait. AB - Gait analysis techniques were used to evaluate the outcome of the femoral derotation osteotomy in children with hemiplegic cerebral palsy. Seventy-one patients were evaluated and classified according to the Winters and Gage scale. Nine of the 13 patients classified as type IV underwent a femoral derotation osteotomy. Postoperatively, all nine patients demonstrated less internal hip rotation, pelvic retraction, and internal foot progression angle. Before surgery patients improve the foot progression by an externally biased hip position within the hip rotation arc. After surgery the hip position is allowed to be more centered within the rotation arc without compromising foot progression. Interestingly, the changed hip position during gait was significantly less than the magnitude of the derotation osteotomy. When contemplating postoperative outcome, consideration of hip position within the arc of rotation and pelvic retraction should be given to avoid undercorrection and residual pelvic compensation with femoral osteotomy based solely on foot progression. PMID- 12724594 TI - Proximal femoral varus rotation osteotomy in cerebral palsy: a prospective gait study. AB - This prospective study examined the kinematic and temporal/spatial effects of proximal femoral varus rotation osteotomy (VRO) on the gait of individuals with cerebral palsy from preoperative to 1-year postoperative status. Participants were a consecutive sample of 37 individuals (14 males, 23 females). The analysis consisted of three-dimensional kinematics, temporal/spatial measures, and functional status. A curve representing the difference between the preoperative and 12-month postoperative conditions was calculated for each joint motion. Ninety-five percent confidence intervals were calculated about the mean difference curves using a bootstrapping technique. VRO resulted in improved cosmesis and objective improvement in gait, including a statistically significant increase in hip external rotation and hip extension, a decrease in anterior pelvic tilt, and an increase in knee extension strength. The use of confidence bands to identify surgical outcomes with respect to kinematic variables has enormous value for patients and professionals. Long-term follow-up is needed to see if the aforementioned gains improve. PMID- 12724595 TI - Gait analysis in low lumbar myelomeningocele patients with unilateral hip dislocation or subluxation. AB - The surgical indications for the treatment of unilateral hip dislocations or subluxations in patients with low lumbar myelomeningocele remain highly debatable. This study examines the influence of unilateral hip dislocation or subluxation on the gait of these patients using three-dimensional gait analysis. Twenty patients with a diagnosis of low lumbar myelomeningocele underwent three dimensional gait analysis. All patients were community ambulators with solid ankle-foot orthoses and crutches who presented with unilateral hip dislocation or subluxation and no scoliosis. The patients were divided in two groups. Group 1 comprised 10 patients who demonstrated either no evidence of hip flexion or adduction contractures or symmetric hip contractures. Group 2 comprised 10 patients with unilateral hip flexion and/or adduction contractures. Pelvic and hip kinematics were assessed to determine the symmetry of motion between the involved and the noninvolved side during walking. Seven patients from group 1 walked with a symmetric gait pattern; only two patients from group 2 walked with a symmetric pattern. Gait symmetry corresponded to the absence of hip contractures or bilateral symmetrical hip contractures and had no relation to the presence of hip dislocation. The authors concluded that reduction of the hip is unnecessary. PMID- 12724597 TI - Pavlik: the man and his method. AB - Because of the high rate of avascular necrosis, Pavlik developed his harness and method of treating developmental dysplasia of the hip in infants. This historical review highlights Pavlik's career from the time he worked with Frejka and then later in his own clinic in Olomouc, Czech Republic (1938). In the 1950s, he wrote five articles on the results of his harness use in developmental dysplasia of the hip. Subsequently, Pavlik's harness has become the orthosis of choice worldwide for treatment of infants with hip dysplasia because of the principles he espoused. PMID- 12724596 TI - Acquired atlantoaxial instability in children with spastic cerebral palsy. AB - The development of nontraumatic atlantoaxial instability in children with spastic cerebral palsy has not been reported. The authors present three patients with severe spastic quadriplegia who developed C1-C2 instability and cervical myelopathy at mean age 12.6 years. These patients demonstrated a similar clinical picture with symptoms attributed to cervical myelopathy in varied severity including apneic episodes, opisthotonus, alteration in muscle tone, torticollis, respiratory problems, hyperreflexia, and bradycardia. Patient 1 was scheduled for surgery but died due to an apneic episode. Patient 2 refused surgery and has been followed for 3 years while his neurologic condition remains unchanged. Patient 3 underwent occipitocervical decompression and fusion, recovered neurologically, and resumed his previous functional skills. Patients demonstrating considerable functional deterioration or insidious change in their established neurologic status should undergo detailed screening to rule out developing upper cervical instability. Early surgical intervention consisting of spinal decompression and fusion may prevent the development of myelopathy. PMID- 12724598 TI - New technique for early screening of developmental dysplasia of the hip: pilot study. AB - An acoustical technique has been developed for early screening of developmental dysplasia of the hip (DDH) in neonates by comparing the sound transmitted across the hips while a vibratory force was applied to the sacrum. The baseline for 90 normal neonates has been established and tests in the frequency bands of 200, 250, and 315 Hz were found to be most effective, achieving high coherence and smallest discrepancy. Sixteen patients with unilateral DDH were examined and the results suggested that coherence below 0.8 in at least one of these frequency bands was strongly indicative of structural asymmetry between both hips, and there was a significant difference between normal neonates and patients with unilateral DDH. By setting the cut-off discrepancy at 2.0 dB, the best sensitivity (100%) would be achieved, and this could be proposed as the threshold for wide-scale screening of DDH. PMID- 12724599 TI - Splintage in developmental dysplasia of the hip: how low can we go? AB - The indications for using abduction splints in developmental dysplasia of the hip (DDH) are not clearly defined. In this study, the authors prospectively evaluated 797 babies born between 1996 and 1998 as part of a limited targeted ultrasound screening program. In the first group (1996-97), babies with clinical hip instability at first scan were placed in a Pavlik harness. In the second group (1998), only babies who had persistent instability at 2 weeks were splinted. Children with persistent major dysplasia at 9 weeks were splinted in both groups. The splintage rate was 1.6/1,000 live births in the first group and 0.8/1,000 live births in the second group. The rate of surgery for DDH did not increase despite a significant decrease in the number of babies being splinted. This study demonstrates that a splintage rate as low as 1.3/1,000 live births (average rate for 1996-98) can be achieved without adversely affecting the outcomes following treatment of DDH. PMID- 12724600 TI - Computed tomographic findings of osteochondritis dissecans following Legg-Calve Perthes disease. AB - Routine radiographs in the follow-up study of Legg-Calve-Perthes disease patient have generally been plain radiographs of the anteroposterior and frog-leg lateral view. Recently, several reports have found that osteochondritis dissecans can develop in 2% to 4% of Legg-Calve-Perthes disease patients. Early roentgenographic recognition of this lesion is very important for its long-term prognosis. However, follow-up examination of the lesion using plain radiography is sometimes not satisfactory in terms of delineating the lesion. Accordingly, the authors performed computed tomography and three-dimensional reconstruction in 13 hips with osteochondritis dissecans following Legg-Calve-Perthes disease and were able to obtain more precise information on the extent of the involvement, the degree of healing, the stability of the osteochondral fragment, and the location of the dislocated loose body. PMID- 12724601 TI - A new innominate osteotomy in Perthes' disease. AB - The purpose of this study was to evaluate the results of the authors' modification of Salter innominate osteotomy in Perthes' disease. The operation was performed in 16 hips of 15 Perthes' patients (11 boys and 4 girls). The posterior half of the ilium was cut by a Gigli saw using a conventional method. The anterior half was osteotomized using a reciprocal saw in a direction of 45 degrees obliquely to the coronal plane and 30 degrees to 40 degrees posteroinferiorly to the sagittal plane. Stable interposition of the bone block was achieved within the osteotomy site. Bone union occurred in all cases within 4 months without displacement of the osteotomy. The mean of the center-edge angle improved from a mean 19 degrees preoperatively to 29.4 degrees postoperatively, and the average epiphyseal extrusion improved from 23% to 9.5%. The modified Salter osteotomy provided much better stability by simply changing the direction of osteotomy. PMID- 12724602 TI - Significance of laboratory and radiologic findings for differentiating between septic arthritis and transient synovitis of the hip. AB - Although significant differences exist in the methods of treatment and prognoses of septic coxitis and transient synovitis in children complaining of acute hip pain, similar symptoms are present in these two diseases at the early stages, and differential diagnosis is difficult. To differentiate between these two diseases, the authors evaluated the clinical, serologic, and radiologic findings and tried to determine factors that could be used as diagnostic criteria. The authors performed a retrospective study by evaluating medical records, plain hip radiographs, and clinical findings in 97 patients with transient synovitis and 27 patients with septic arthritis. Univariate analysis showed significant differences in body temperature, serum WBC count, and ESR and CRP levels of the two patient groups. Plain radiographs showed a displacement or blurring of periarticular fat pads in all patients with acute septic arthritis, and multivariate regression analysis showed that body temperature >37 degrees C, ESR >20 mm/h, CRP >1 mg/dL, WBC >11,000/mL, and an increased hip joint space of >2 mm were independent multivariate predictors of acute septic arthritis. The authors conclude that the independent multivariate predictors are effective indices for the differential diagnosis of acute septic coxitis and transient synovitis. PMID- 12724603 TI - Assessment of the test characteristics of C-reactive protein for septic arthritis in children. AB - The purpose of this study was to determine the test characteristics of C-reactive protein (CRP) in the diagnosis of septic arthritis in children and to compare with erythrocyte sedimentation rate (ESR). The authors reviewed patients with synovial fluid aspiration sent for culture and Gram stain for whom a CRP was drawn within 24 hours of presentation. Descriptive statistics and univariate analyses were performed. Results for CRP were compared with ESR. Thirty-nine of 133 patients had septic arthritis. Sensitivity of CRP ranged from 41% to 90%, specificity from 29% to 85%. Positive predictive values ranged from 34% to 53%, negative predictive values from 78% to 87%. In comparison to ESR, CRP is a better independent predictor of disease. CRP is a better negative predictor than a positive predictor of disease. Indeed, if the CRP is <1.0 mg/dL, the probability that the patient does not have septic arthritis is 87%. PMID- 12724604 TI - Treatment of leg length discrepancy with temporary epiphyseal stapling in children with juvenile idiopathic arthritis during 1957-99. AB - Temporary epiphyseal stapling is a flexible method for correction of leg length discrepancy. Due to a high risk of premature epiphyseal plate closure and other complications reported in previous studies, the technique has not been in wide use. In this study a retrospective analysis of 71 knees in patients with juvenile idiopathic arthritis was performed to determine the safety and effectiveness of the method. Only seven minor reversible complications were encountered, and the authors found the method suitable even for immunologically compromised patients. PMID- 12724605 TI - Leg length inequality and epiphysiodesis: review of 96 cases. AB - A retrospective analysis of 96 patients who underwent an epiphysiodesis procedure for leg length discrepancy was performed. Forty patients were identified who had an open Phemister-type epiphysiodesis. Fifty-six patients underwent a percutaneous epiphysiodesis procedure. All patients' medical records and radiographs were reviewed for complications. No angular deformities or epiphysiodesis failures occurred in the open group; however, one deep infection requiring intravenous antibiotics and serial surgical debridement was successfully treated. Five complications occurred in the percutaneous group. The complications included two superficial infections, two failures of physeal arrest, and one postoperative angular deformity. The occurrence of complications in the two groups was not statistically significant. Regular follow-up and radiographic evaluation of patients who have undergone a percutaneous epiphysiodesis is essential. Failure of physeal arrest and angular deformities are rare in this patient population, but they can occur. PMID- 12724606 TI - Adjustable bracing technique for the prevention of knee flexion contracture during tibial lengthening. AB - The authors present a simple thigh-knee brace that prevents flexion contracture during tibial lengthening. The brace is strapped to the thigh and connected to the Ilizarov frame via two simple hinges. While in the brace the knee can be mobilized for physiotherapy and locked in extension during rest. PMID- 12724607 TI - Correction of tibia vara with six-axis deformity analysis and the Taylor Spatial Frame. AB - Operative correction for infantile and adolescent tibia vara has been described using both external and internal fixation. Gradual correction using a circular fixator offers the advantage of accurate coronal, sagittal, and axial plane correction without significant soft tissue dissection. This study evaluated the use of six-axis deformity analysis and the Taylor Spatial Frame (TSF) for the correction of tibia vara. Nineteen patients (22 tibias), 6 with infantile and 13 with adolescent tibia vara, underwent correction with TSF. On the basis of mechanical axis correction, 21 of 22 tibias were corrected within 3 degrees of normal. Using Schoenecker's criteria, all patients achieved good results (no pain, <5 degrees difference in tibial-femoral angle from the normal side). Complications included one intractable pin-site infection, two superficial pin site infections, and one delayed union. Six-axis deformity analysis and TSF provide accurate and safe correction of infantile and adolescent tibia vara. PMID- 12724608 TI - Surgical management of chronic lateral ankle instability in adolescents. AB - Chronic lateral ankle instability in adolescents is an infrequent result of either an acute or recurrent ankle inversion injury. The majority of these patients improve with nonoperative management. Numerous surgical interventions have been proposed when nonoperative management is ineffective. Since 1979, 12 adolescents, all girls, have undergone surgical treatment of chronic lateral ankle instability at a major pediatric center. The average age at the time of surgery was 14 years and 3 months. Questionnaires were sent to each patient to assess subjective long-term outcome. Follow-up ranged from 1 year and 5 months to 8 years and 5 months (average 3 years and 1 month). The Watson-Jones technique was performed in six, the Evans in four, the Chrisman-Snook in one, and the Brostrom in one. At most recent follow-up, 11 adolescents had a stable ankle, 9 had a full range of ankle and subtalar motion, and 6 had returned to all activities. PMID- 12724609 TI - Surgical technique and preliminary results of a new fixation concept for olecranon fractures in children. AB - The authors report preliminary results of an original fixation technique used for the treatment of olecranon fractures in six children. This technique uses two threaded pins fitted with an adjustable lock, introduced through a minimal skin incision. With a mean follow-up of 14 months, clinical results are excellent in five cases and good in one case. Radiologic results are satisfying, with five anatomic reductions and without any secondary displacement. No growth impairment has been observed, but the follow-up period is still too short to conclude that this technique is superior, as far as this factor is concerned. However, the simplicity of the technique and its efficiency have led the authors to use it routinely for this fracture type. PMID- 12724611 TI - Pediatric hand friction burn injuries secondary to treadmills. AB - A characteristic hand friction burn secondary to a treadmill fitness machine has been seen in fourteen pediatric patients. This retrospective review looks at mechanism of injury, digit(s) involved, depth of burn, initial treatment, time to referral and its effect on the final result, and the need and results of definitive treatment. Based on our findings, certain recommendations are made to avoid potential sequela due to this hand friction burn. PMID- 12724610 TI - Operative management of pediatric scaphoid fracture nonunion. AB - Fractures of the scaphoid are uncommon childhood injuries that can usually be successfully managed by standard immobilization techniques. However, scaphoid waist fractures are less frequent in children and may proceed to nonunion. The authors retrospectively reviewed the charts and radiographs of 19 adolescents with 20 scaphoid waist fractures that had required operative intervention for nonunion from January 1975 to December 2000. All cases had undergone a trial of plaster immobilization, averaging 4.2 months, before operative intervention was performed. Operative management resulted in a 100% rate of union. Herbert or AO screw fixation and grafting was performed in 11, 2 had K-wire fixation and bone grafting, 6 had bone grafting and no fixation, and one had screw fixation with no graft. All but three patients were available for telephone interview, with an average follow-up of 6.1 years. One reported having mild pain at rest, 4 had discomfort with heavy activity, and the remaining 12 were pain-free. Only two had restriction of activity because of pain, with the remainder returning to previous levels of function. All but one reported complete satisfaction with the results of their operation. Operative intervention of nonunion of scaphoid waist fractures in children consistently offers successful fusion, with very low rates of nonunion and a high rate of patient satisfaction. PMID- 12724612 TI - Clubfoot salvage: a review of the past decade's contributions. PMID- 12724613 TI - Surgical prevention of foot deformity in patients with Duchenne muscular dystrophy. PMID- 12724614 TI - Prothrombotic genetic risk factors and heparin-induced thrombocytopenia. PMID- 12724615 TI - Pharmacogenetics of oral anticoagulants. AB - There is wide interindividual variation in oral anticoagulant dose requirement, which is partly genetically determined. Several cytochrome P450s contribute to oxidative metabolism of oral anticoagulants. The most important of these is CYP2C9, which hydroxylates the S-enantiomers of warfarin, acenocoumarol and phenprocoumon with high catalytic activity. In at least eight separate clinical studies, possession of the CYP2C9*2 or CYP2C9*3 variant alleles, which result in decreased enzyme activity, has been associated with a significant decrease in a mean warfarin dose requirement. Several studies also suggest that possession of a CYP2C9 variant allele is associated with an increased risk of adverse events, such as bleeding. Possession of the CYP2C9*3 variant also appears to be associated with a low acenocoumarol dose requirement. Other genetic factors, such as polymorphisms in the cytochromes P450 that metabolize the R-enantiomers of warfarin and acenocoumarol, may also be relevant to anticoagulant dose. The molecular basis of anticoagulant resistance where a higher than normal dose of anticoagulant is required remains unclear, but could be due to unusually high CYP2C9 activity (pharmacokinetic resistance) or to an abnormality in the target enzyme vitamin K epoxide reductase (pharmacodynamic resistance). PMID- 12724616 TI - Platelet receptor and clotting factor polymorphisms as genetic risk factors for thromboembolic complications in heparin-induced thrombocytopenia. AB - Heparin-induced thrombocytopenia (HIT) is an immune mediated adverse reaction to heparin treatment often associated with limb- and/or life-threatening thromboembolic complications (TECs). Presently, no prognostic marker has been identified that allows differentiation between mildly (isolated thrombocytopenia) and severely (TECs) affected patients. This study assesses the impact of platelet glycoprotein- and clotting factor polymorphisms in HIT-patients with isolated thrombocytopenia compared to HIT-patients with TECs. Sixty-three HIT-patients with isolated thrombocytopenia and 79 HIT-patients with HIT-related TECs were genotyped for GPIIb-IIIa polymorphisms (HPA-1, HPA-3), GPIa-IIa polymorphisms (HPA-5, GPIaC807T), GPIb-IX-V polymorphisms (HPA-2, Kozak-5, VNTR), and clotting factor polymorphisms (FV-Leiden R506Q, prothrombin PT-G20210A and MTHFR C677T). Women more often presented with TECs than men (P = 0.04). No differences in genotype frequencies could be seen on comparing HIT-patients with and without TECs. Analysing men and women separately, the C allele of the Kozak polymorphism was overrepresented in men who developed TECs (P = 0.034). The enhanced risk of women to develop HIT-associated TECs remains unexplained but it is potentially important in view of recent data on sex-hormone related changes of haemostasis. There was no correlation between platelet glycoprotein- and clotting factor polymorphisms and the risk to develop HIT-associated TECs. An association between the development of TECs and the Kozak-5C allele could be seen among male patients. However, this would need to be assessed in further larger studies. Most likely, the high levels of thrombin generation during acute HIT are so procoagulant that less pronounced risk factors such as polymorphisms are overshadowed. PMID- 12724617 TI - Polymorphism in the P-glycoprotein drug transporter MDR1 gene: a possible link between environmental and genetic factors in Parkinson's disease. AB - P-glycoprotein is a membrane protein encoded by the MDR1 gene, which demonstrates functional polymorphism. It is present in endothelial cells of the blood-brain barrier, thus limiting accumulation of its substrates in the central nervous system. Many epidemiological studies suggest an association between pesticides, which are substrates for P-glycoprotein, and Parkinson's disease. It was hypothesized that polymorphism of the MDR1 gene could modulate interindividual susceptibility for the disease in subjects exposed to pesticides. In a pilot case control study involving 107 Parkinson's disease patients (30 early onset and 77 late onset patients; 59 exposed to pesticides and 48 non-exposed) and 103 controls, C3435T polymorphism of the gene was analysed. No statistically significant correlation between MDR1 gene polymorphism and Parkinson's disease was found. The 3435TT genotype was noted more frequently, but not significantly, in patients with early onset compared to late onset disease (23.3% versus 10.4%, respectively). A significant association between patients with parkinsonism exposed to pesticides and C3435T polymorphism of the MDR1 gene was found. Comparing the exposed and non-exposed patients, a statistically higher frequency of heterozygous subjects was observed (72.9% versus 47.9%, respectively). This genotype was associated with a significant, almost three-fold increased risk of disease. Similarly, a higher frequency of 3435TT subjects was revealed in exposed subjects (15.5%) compared to non-exposed patients (12.5%). In exposed versus non exposed subjects, patients carrying at least one 3435T allele (i.e. homozygous and heterozygous) had a significant, five-fold higher risk of Parkinson's disease. Thus, it appears that mutation of the MDR1 gene predisposes to damaging effects of pesticides, and possibly other toxic xenobiotics transported by P glycoprotein, leading to Parkinson's disease. PMID- 12724618 TI - Frequency of butyrylcholinesterase gene mutations in individuals with abnormal inhibition numbers: an Italian-population study. AB - OBJECTIVES: More than 30 genetic variants of serum cholinesterase (butyrylcholinesterase, BChE) have been described. Some of them (the atypical and the fluoride-resistant variants) are well known because carriers are prone to develop prolonged apnea following the administration of the muscle relaxant succinylcholine. Genotype characterization is therefore important in order to prevent such episodes. Genetic studies have so far focused on selected individuals or families rather than on the random population. METHODS: From a large group of healthy blood donors (n = 2609), we selected all the 58 individuals with low serum cholinesterase activity: among them 28 subjects had abnormal dibucaine and fluoride inhibition numbers. Twenty-five mutations in the coding region of the human cholinesterase gene were analyzed. RESULTS: All individuals with abnormal inhibition numbers were homozygotes or double heterozygotes in several mutations. Asp70Gly (Atypical variant) and Ala539Thr (K variant) were the most frequently observed amino acid substitutions. The majority of subjects with low BChE activity but normal dibucaine and fluoride number presented only the K form. We analyzed 106 randomly chosen subjects for K and atypical variants. Carriers of these alleles were at risk of low BChE activity (OR = 9.55, 95%CI, 5.61-16.26 and OR = 30.33, 95%CI, 7.05-130.52 respectively). CONCLUSIONS: Data obtained from this study help to better define the etiology of low BChE activity and the role of the rather common K allele. It is the first time that such a large population has been screened for so many mutations. BChE is also implicated in detoxifying cocaine; therefore genetic analysis could be useful in cases of cocaine toxicity in Italian subjects. PMID- 12724619 TI - A microsatellite repeat in the promoter of the N-methyl-D-aspartate receptor 2A subunit (GRIN2A) gene suppresses transcriptional activity and correlates with chronic outcome in schizophrenia. AB - Hypofunction of the N-methyl-D-aspartate (NMDA) receptor has been hypothesized to underlie the pathophysiology of schizophrenia, based on the observation that non competitive antagonists of the NMDA receptor, such as phencyclidine, induce schizophrenia-like symptoms. Mice lacking the NR2A subunit of the NMDA receptor complex are known to display abnormal behaviour, similar to schizophrenic symptoms. The expression of NR2A starts at puberty, a period corresponding to the clinical onset of schizophrenia. This evidence suggests that the NR2A (GRIN2A) gene may play a role in the development of schizophrenia and disease phenotypes. In this study, we performed a genetic analysis of this gene in schizophrenia. Analysis of the GRIN2A gene detected four single nucleotide polymorphisms, and a variable (GT)(n) repeat in the promoter region of the gene. A case-control study (375 schizophrenics and 378 controls) demonstrated evidence of an association between the repeat polymorphism and the disease (P = 0.05, Mann-Whitney test), with longer alleles overly represented in patients. An in-vitro promoter assay revealed a length dependent inhibition of transcriptional activity by the (GT)(n) repeat, which was consistent with a receptor binding assay in postmortem brains. Significantly, the score of symptom severity in chronic patients correlated with repeat size (P = 0.01, Spearman's Rank test). These results illustrate a genotype phenotype correlation in schizophrenia and suggest that the longer (GT)(n) stretch may act as a risk-conferring factor that worsens chronic outcome by reducing GRIN2A levels in the brain. PMID- 12724620 TI - Role of the eNOS Glu298Asp variant on the GNB3825T allele dependent determination of alpha-adrenergic coronary constriction. AB - The 825T allele of a polymorphism at GNB3, encoding the G protein beta(3) subunit, is associated with an enhanced coronary blood flow (CBF) reduction in response to alpha(2)-, but not to alpha(1)-adrenoceptor activation. Because the regulation of vascular tone by alpha(2)-adrenoceptors includes direct vasoconstriction as well as vasodilatation by endothelial release of nitric oxide, the eNOS Glu298Asp variant might further contribute to explain the variability of CBF reduction in response to alpha-adrenoceptor activation. Genotyping at the GNB3 and the eNOS gene was performed on 48 individuals receiving either the alpha(1)-adrenoceptor agonist methoxamine (5 mg i.c.) and/or the alpha(2)-adrenoceptor agonist BHT 933 (5 mg i.c.). CBF was calculated from quantitative coronary angiography and intracoronary Doppler flow velocity measurement. To analyse the impact of genotypes and coronary artery disease, a linear regression model was used, including cholesterol levels, heart rate, smoking and mean aortic blood pressure. An initial, univariate analysis suggested an impact of the eNOS Glu298Asp variant on alpha(2)-adrenoceptor-induced coronary constriction (CBF reduction 53.4 ch006.1 TT/TG versus 30.7 ch006.9% GG; P = 0.003). However, multifactorial analysis showed that the GNB3825T allele was associated exclusively with the CBF reduction on alpha(2)-adrenoceptor activation (58.2 +/- 4.4% TT/TC versus 27.9 +/- 4.3% CC; P < 0.0001). Contrary to the initial analysis, the Glu298Asp variant of the eNOS gene provides no additional information on the genetic basis of alpha(2)-adrenoceptor-induced coronary vasoconstriction, which appears exclusively associated to the 825T allele at GNB3. Analysis of modifying genes appears crucial for the understanding of genetic associations. PMID- 12724621 TI - NAT2 slow acetylator function as a risk indicator for age-related cataract formation. AB - OBJECTIVES: To show that the slow arylamine N-acetyltransferase type 2 (NAT2) catalysed acetylator function is associated with the development of age-related cataracts. METHODS: Both the acetylator phenotype and genotype of 139 patients with age-related cataracts were determined, and the distribution of the acetylator subtypes in the case population was compared with the distribution in the general (control) population. The genotype was determined by restriction enzyme analysis of DNA, and the phenotype was determined using the elimination characteristics of isoniazid as discriminant. RESULTS: The frequency of alleles coding for slow acetylator characteristics was higher in the patients than in the controls, and the difference was significant (P = 0.013). CONCLUSIONS: Slow acetylators are at higher risk of developing age-related cataracts than fast acetylators and we suggest that exogenous factors, which can be detoxified by acetylation, are aetiological agents for cataract formation. Identification of and avoidance of such (environmental) agents should reduce the incidence of age related cataracts. PMID- 12724622 TI - Novel paraoxonase (PON1) nonsense and missense mutations predicted by functional genomic assay of PON1 status. AB - Paraoxonase (PON1) has been termed an environmental response enzyme for its function in the detoxification of organophosphate pesticides, nerve agents and pharmaceuticals such as glucocorticoids and statins, as well as its cardioprotective role in breaking down oxidized LDL. PON1(192) genotype can be predicted with high accuracy from an examination of the two-dimensional plot of paraoxon and diazoxon hydrolysis rates [ 1]. Individuals for whom this functional genomic assay failed to predict PON1(192) genotype, or who had a low PON activity relative to others with the same genotype, were predicted to have genetic alterations that explained the inconsistency. Sequencing of the PON1 region of 23 Caucasian individuals detected a nonsense mutation changing amino acid 194 from a Trp to a stop codon (PON1(Trp194stop)). It was predicted that subjects who genotyped as PON1(192QR) but phenotyped as PON1(192QQ) or PON1(192RR) might carry the protein truncation mutation for which the defective product failed to be detected by the phenotyping assay. Screening of the five discordant subjects resulted in the detection of a single Caucasian carrying the stop codon, and determined its phasing on the PON1(192R) allele. Sequencing confirmed the change and revealed an additional subject with a likely deletion of the 5' end of the PON1 gene. Additional sequencing of 25 subjects with low PON1 activities identified two additional previously undescribed PON1 mutations, which may affect PON1 function: PON1(Pro90Leu) associated with the PON1(192Q) allele and PON1(Asp124missplice) associated with the PON1(192R) allele. PMID- 12724623 TI - Functional characterization in yeast of genetic variants in the human equilibrative nucleoside transporter, ENT1. AB - The human equilibrative nucleoside transporter, ENT1, appears to play a critical role in the disposition of nucleosides and nucleoside analogs used clinically as anti-viral and anti-cancer drugs. Recently, we identified variants of ENT1 in an ethnically diverse DNA sample set from 247 individuals, focusing primarily on the coding region. The goal of the present study was to analyse the haplotype structure and functionally characterize the variants of ENT1. We observed that a single haplotype, ENT1*1, accounted for 91.3% of the 494 chromosomes. Functional analysis in Saccharomyces cerevisiae revealed no differences in the kinetics of uptake of nucleosides and nucleoside analogs by the two non-synonymous variant transporters, ENT1-I216T and ENT1-E391K, and the reference ENT1. These results, together with the observation that there are few haplotypes of ENT1, indicate that coding region variants of ENT1 do not contribute to inter-individual differences in response to nucleoside analog drugs. PMID- 12724625 TI - Why pediatric surgery? A personal journey through the first 50 years. PMID- 12724626 TI - Extracapsular extension of the sentinel lymph node metastasis: a predictor of nonsentinel node tumor burden. AB - OBJECTIVE: To identify predictors of nonsentinel node (NSN) tumor involvement in patients with a tumor-involved sentinel node (SN). SUMMARY BACKGROUND DATA: For many breast cancer patients who undergo intraoperative lymphatic mapping and sentinel lymphadenectomy (LM/SL), the SN is the only tumor-involved axillary node. Associations between NSN tumor involvement and several clinical and histopathologic factors have been identified. The authors hypothesize that extracapsular extension (ECE) of the SN metastasis is highly predictive of NSN tumor involvement. METHODS: Between May 1998 and December 2001, 260 patients (263 cases) with clinical T1 or T2 (<5.0 cm) breast cancer underwent LM/SL at the University of North Carolina, using a combined blue dye and technetium sulfur colloid technique. In all cases with a tumor-involved SN, axillary lymph node dissection (ALND) was recommended. Statistical analysis, with Pearson chi-square tests, Fisher exact test, and multiple logistic regression, was performed. RESULTS: The SN contained tumor in 74 (28.1%) cases. ALND was performed in 70 of the 74 cases. ECE of the SN metastasis was present in 18 (25.7%) of the 70 cases. Patients with ECE of the SN metastasis were more likely to have NSN tumor involvement and had a greater total number of tumor-involved nodes than patients without ECE of the SN metastasis. Increasing size of the SN metastasis and increasing size of the primary tumor, examined as continuous variables, were associated with an increased likelihood of NSN tumor involvement on univariate analysis. However, only ECE of the SN metastasis was associated with NSN tumor involvement on multivariate analysis. CONCLUSIONS: ECE of the SN metastasis is a strong predictor of NSN tumor involvement. All patients with ECE of the SN metastasis should undergo mandatory completion ALND. PMID- 12724627 TI - Duct drainage alone is sufficient in the operative management of pancreatic pseudocyst in patients with chronic pancreatitis. AB - OBJECTIVE: To test a hypothesis that definitive management of pseudocyst associated with chronic pancreatitis is predicated on addressing pancreatic ductal anatomy. SUMMARY BACKGROUND DATA: The authors have previously confirmed the impact of pancreatic ductal anatomic abnormalities on the success of percutaneous drainage of pancreatic pseudocyst. The authors have further defined a system to categorize the pancreatic ductal abnormalities that can be seen with pancreatic pseudocyst. The authors have published, as have others, the usefulness of defining ductal anatomy when managing pancreatic pseudocysts associated with chronic pancreatitis. METHODS: Beginning in 1985, all patients with pseudocyst who were candidates for intervention (operative, percutaneous, or endoscopic) have undergone endoscopic retrograde cholangiopancreatography (ERCP). An associated diagnosis of chronic pancreatitis was established by means of ERCP findings. Patients were candidates for longitudinal pancreaticojejunostomy (LPJ) if they had a pancreatic ductal diameter greater than 7 mm. In a nonrandomized fashion, patients were managed with either combined simultaneous LPJ and pseudocyst drainage or with LPJ alone. RESULTS: Two hundred fifty-three patients with pseudocyst have been evaluated. Among these there have been 103 patients with chronic pancreatitis and main pancreatic duct (MPD) dilatation (>7 mm). Among these 103 patients, 56 underwent combined LPJ/pseudocyst drainage and 47 had LPJ alone. Compared to combined LPJ/pseudocyst drainage, the patients undergoing LPJ alone had a shorter operative time, slightly less transfusion requirement, slightly reduced length of hospital stay, and slightly reduced complication rate. Long-term pain relief was achieved in 90%, and pseudocyst recurrence was less than 1%. Rates of each of these long-term outcomes were nearly incidental among the two groups. CONCLUSIONS: Ductal drainage alone (LPJ) is sufficient in patients with chronic pancreatitis (MPD > 7 mm) and an associated pseudocyst. Simultaneous drainage of pseudocyst is not necessary. PMID- 12724628 TI - Abdominal aortic aneurysms in "high-risk" surgical patients: comparison of open and endovascular repair. AB - OBJECTIVE: To evaluate the early results of endovascular grafting for high-risk surgical candidates in the treatment of abdominal aortic aneurysms (AAA). SUMMARY BACKGROUND DATA: Since the approval of endoluminal grafts for treatment of AAA, endovascular repair of AAA (EVAR) has expanded to include patients originally considered too ill for open AAA repair. However, some concern has been expressed regarding technical failure and the durability of endovascular grafts. METHODS: The University of Alabama at Birmingham (UAB) Computerized Vascular Registry identified all patients who underwent abdominal aneurysm repair between January 1, 2000, and June 12, 2002. Patients were stratified by type of repair (open AAA vs. EVAR) and were classified as low risk or high risk. Patients with at least one of the following classifications were classified as high risk: age more than 80 years, chronic renal failure (creatinine > 2.0), compromised cardiac function (diminished ventricular function or severe coronary artery disease), poor pulmonary function, reoperative aortic procedure, a "hostile" abdomen, or an emergency operation. Death, systemic complications, and length of stay were tabulated for each group. RESULTS: During this 28-month period, 404 patients underwent AAA repair at UAB. Eighteen patients (4.5%) died within 30 days of their repair or during the same hospitalization. Two hundred seventeen patients (53%) were classified as high risk. Two hundred fifty-nine patients (64%) underwent EVAR repair, and 130 (50%) of these were considered high-risk patients (including four emergency procedures). One hundred forty-five patients (36%) underwent open AAA repair, including 15 emergency operations. All deaths occurred in the high-risk group: 12 (8.3%) died after open AAA repair and 6 (2.3%) died after EVAR repair. Postoperative length of stay was shorter for EVAR repair compared to open AAA. CONCLUSIONS: High-risk and low-risk patients can undergo EVAR repair with a lower rate of short-term systemic complications and a shorter length of stay compared to open AAA. Despite concern regarding the durability of EVAR, high-risk patients should be evaluated for EVAR repair before committing to open AAA repair. PMID- 12724629 TI - Use of the Jarvik 2000 left ventricular assist system as a bridge to heart transplantation or as destination therapy for patients with chronic heart failure. AB - OBJECTIVE: To evaluate the Jarvik 2000 axial flow left ventricular assist system (LVAS) as a bridge to transplant and as destination therapy. SUMMARY BACKGROUND DATA: The Jarvik 2000 LVAS was implanted in 22 patients (16 men, 6 women; mean age 53 years) as a bridge to transplant (in the United States) and in 4 patients (all men; mean age 62.8 years) as destination therapy (in the United Kingdom). All patients in both of these initial feasibility studies were in NYHA class 4. METHODS: The pump was implanted through a thoracotomy or median sternotomy incision with the aid of partial cardiopulmonary bypass in bridge-to-transplant patients. A skull-mounted percutaneous power delivery was used for the patients who received the pump as destination therapy. RESULTS: Of the 22 bridge-to transplant patients, 13 underwent transplant; 7 died during support; and 2 studies are ongoing. The surviving patients have an average follow-up of 15 months; one died at 2.6 months after transplant, and the remaining patients are all in NYHA class 1. Support averaged 67.1 days. Deaths were due to acute myocardial infarction in two patients and multiorgan failure in five patients. Hemodynamic function improved with LVAS support. The average cardiac index increased 70.6% by 48 hours after implant, pulmonary capillary wedge pressure decreased 44%, systemic vascular resistance decreased significantly, and inotropic support became unnecessary. Similar results have been seen in the patients who received the device as destination therapy. In that series, one patient died of subdural hematoma 380 days after implant. The other two patients are in NYHA class 1, 642 and 889 days after implant. The average cardiac index increased 89.5%, and pulmonary capillary wedge decreased 52.2%. CONCLUSIONS: The Jarvik 2000 axial-flow LVAS can be used safely in selected patients to provide support until transplant or as destination therapy. In this series, the patients who most benefited from this device were those who required true left ventricular assistance rather than total capture of left ventricular output. Current experience indicates that continuous offloading of the ventricle is most effective when there is enough residual myocardial function to maintain pulsatility and aortic root ejection and to maintain, with nonpulsatile pump support, a normal cardiac index as well as reinstitution of the Frank-Starling response to the native ventricle. PMID- 12724630 TI - Paradigm shift in the management of gastroesophageal reflux disease. AB - OBJECTIVE: To compare the short-term results of the radiofrequency treatment of the gastroesophageal junction known as the Stretta procedure versus laparoscopic fundoplication (LF) in patients with gastroesophageal reflux disease (GERD). SUMMARY BACKGROUND DATA: The Stretta procedure has been shown to be safe, well tolerated, and highly effective in the treatment of GERD. METHODS: All patients presenting to Vanderbilt University Medical Center for surgical evaluation of GERD between August 2000 and March 2002 were prospectively evaluated under an IRB approved protocol. All patients underwent esophageal motility testing and endoscopy that documented GERD preoperatively, either by a positive 24-hour pH study or biopsy-proven esophagitis. Patients were offered the Stretta procedure if they had documented GERD and did not have a hiatal hernia larger than 2 cm, LES pressure less than 8 mmHg, or Barrett's esophagus. Patients with larger hiatal hernias, LES pressure less than 8 mmHg, or Barrett's were offered LF. All patients were studied pre- and postoperatively with validated GERD-specific quality-of-life questionnaires (QOLRAD) and short-form health surveys (SF-12). Current medication use and satisfaction with the procedure was also obtained. RESULTS: Results are reported as mean +/- SEM. Seventy-five patients (age 49 +/- 14 years, 44% male, 56% female) underwent LF and 65 patients (age 46 +/- 12 years, 42%, 58% female) underwent the Stretta procedure. Preoperative esophageal acid exposure time was higher in the LF group. Preoperative LES pressure was higher in the Stretta group. In the LF group, 41% had large hiatal hernias (>2 cm), 8 patients required Collis gastroplasty, 6 had Barrett's esophagus, and 10 had undergone previous fundoplication. At 6 months, the QOLRAD and SF-12 scores were significantly improved within both groups. There was an equal magnitude of improvement between pre- and postoperative QOLRAD and SF-12 scores between Stretta and LF patients. Fifty-eight percent of Stretta patients were off proton pump inhibitors, and an additional 31% had reduced their dose significantly; 97% of LF patients were off PPIs. Twenty-two Stretta patients returned for 24-hour pH testing at a mean of 7.2 +/- 0.5 months, and there was a significant reduction in esophageal acid exposure time. Both groups were highly satisfied with their procedure. CONCLUSIONS: The addition of a less invasive, endoscopic treatment for GERD to the surgical algorithm has allowed the authors to stratify the management of GERD patients to treatment with either Stretta or LF according to size of hiatal hernia, LES pressure, Barrett's esophagus, and significant pulmonary symptoms. Patients undergoing Stretta are highly satisfied and have improved GERD symptoms and quality of life comparable to LF. The Stretta procedure is an effective alternative to LF in well-selected patients. PMID- 12724631 TI - Possible primary lymph node gastrinoma: occurrence, natural history, and predictive factors: a prospective study. AB - OBJECTIVE: To analyze the results of a prospective study of 176 patients with Zollinger-Ellison syndrome (ZES) (138 sporadic, 38 MEN1) undergoing 207 operations over a 17-year period. SUMMARY BACKGROUND DATA: The existence of lymph node (LN) primary gastrinoma causing ZES is controversial. METHODS: Three groups of patients were compared: LN only resected, cured, and no relapse (likely LN primary); same criteria but relapse (unlikely LN primary); and duodenal primary and LN metastases (Duo-LN). RESULTS: Forty-five (26%) had only LN(s) as the initial tumor found. Twenty-six of the 45 (58%) fit the definition of a likely LN primary because they were apparently cured postresection. At 10.4 +/- 1.2 years, 69% of the 26 patients with likely LN primary tumors have remained cured and have LN primaries. In the 8 of 26 with recurrent ZES, it occurred at 5 +/- 1 years, and 3 had duodenal gastrinoma that had been missed. Ten percent (13/138) of all patients with sporadic ZES and 0% (0/38) with ZES and MEN1 remained cured with only a LN tumor removed. In patients with sporadic gastrinomas no clinical, laboratory, or radiographic localization feature differed among patients with likely LN primary (n = 16) and those with unlikely LN primary (n = 6) or those with Duo-LN (n = 37). In the likely LN primary group, the largest LN was 2.2 +/- 0.2 cm, the number of LNs removed was 1.3 +/- 0.1 (25% > or =1 LN), and 78% were in the gastrinoma triangle, which also did not differ from the other 2 groups. Disease-free survival was similar in the likely LN primary group, patients with Duo-LN, and those with pancreatic primaries. CONCLUSIONS: These results support the conclusion that primary LN gastrinomas occur and are not rare (approximately 10% of sporadic cases). These results suggest that a proportion (25%) of these tumors are either multiple or malignant. Because no clinical, laboratory, or tumoral characteristic distinguishes patients with LN primary tumors, all patients with ZES undergoing surgery should have an extensive exploration to exclude duodenal or pancreatic tumors and routine removal of lymph nodes in the gastrinoma triangle. PMID- 12724632 TI - Characterization of renal allograft rejection by urinary proteomic analysis. AB - OBJECTIVE: To develop a diagnostic method with no morbidity or mortality for the detection of acute renal transplant rejection. SUMMARY BACKGROUND DATA: Rejection constitutes the major impediment to the success of transplantation. Currently available methods, including clinical presentation and biochemical organ function parameters, often fail to detect rejection until late stages of progression. Renal biopsies have associated morbidity and mortality and provide only a limited sample of the organ. METHODS: Thirty-four urine samples were collected from 32 renal transplant patients at various stages posttransplantation. Samples were collected from 17 transplant recipients with acute rejection and 15 patients with no rejection. Samples from patients less than 4 days posttransplant were omitted from data analysis due to the presence of excessive inflammatory response proteins. Rejection status was confirmed by kidney biopsy. Specimens were analyzed in triplicate using SELDI mass spectrometry. The obtained spectra were subjected to bioinformatic analysis using ProPeak as well as CART (Classification and Regression Tree) algorithms to identify rejection biomarker candidates. These candidates were identified by their molecular weight and ranked by their ability to distinguish between nonrejection and rejection based on receiver operating characteristic (ROC) analysis. The candidates with the highest area under the ROC curve (AUC) exhibited the best diagnostic performance. RESULTS: The best candidate biomarkers demonstrated highly successful diagnostic performance: 6.5 kd (AUC = 0.839, P <.0001), 6.7 kd (AUC = 0.839, P <.0001), 6.6 kd (AUC = 0.807, P <.0001), 7.1 kd (AUC = 0.807, P <.0001), and 13.4 kd (AUC = 0.804, P <.0001). A separate analysis using the CART algorithm in the Ciphergen Biomarker Pattern Software correctly classified 91% of the 34 specimens in the training set, giving a sensitivity of 83% and specificity of 100% using two separate biomarker candidates at 10.0 kd and 3.4 kd. CONCLUSIONS: Biomarker candidates exist in urine that have the ability to distinguish between renal transplant patients with no rejection and those with acute rejection. These biomarker candidates are the basis for development of a noninvasive method of diagnosing acute rejection without the morbidity and mortality associated with needle biopsy. The combination of biomarkers into a panel for diagnosis leads to the possibility of enhanced diagnostic performance. PMID- 12724633 TI - Liver transplantation for fulminant hepatic failure: experience with more than 200 patients over a 17-year period. AB - OBJECTIVE: To analyze outcomes after liver transplantation (LT) in patients with fulminant hepatic failure (FHF) with emphasis on pretransplant variables that can potentially help predict posttransplant outcome. SUMMARY BACKGROUND DATA: FHF is a formidable clinical problem associated with a high mortality rate. While LT is the treatment of choice for irreversible FHF, few investigations have examined pretransplant variables that can potentially predict outcome after LT. METHODS: A retrospective review was undertaken of all patients undergoing LT for FHF at a single transplant center. The median follow-up was 41 months. Thirty-five variables were analyzed by univariate and multivariate analysis to determine their impact on patient and graft survival. RESULTS: Two hundred four patients (60% female, median age 20.2 years) required urgent LT for FHF. Before LT, the majority of patients were comatose (76%), on hemodialysis (16%), and ICU-bound. The 1- and 5-year survival rates were 73% and 67% (patient) and 63% and 57% (graft). The primary cause of patient death was sepsis, and the primary cause of graft failure was primary graft nonfunction. Univariate analysis of pre-LT variables revealed that 19 variables predicted survival. From these results, multivariate analysis determined that the serum creatinine was the single most important prognosticator of patient survival. CONCLUSIONS: This study, representing one of the largest published series on LT for FHF, demonstrates a long-term survival of nearly 70% and develops a clinically applicable and readily measurable set of pretransplant factors that determine posttransplant outcome. PMID- 12724634 TI - Enteral feeding preserves mucosal immunity despite in vivo MAdCAM-1 blockade of lymphocyte homing. AB - OBJECTIVE: To determine the influence of route of nutrition on gut mucosal addressin cellular adhesion molecule-1 (MAdCAM-1) expression and the effect of MAdCAM-1 blockade on gut-associated lymphoid tissue (GALT) lymphocyte populations and established respiratory antibacterial immunity. SUMMARY BACKGROUND DATA: Lymphocytes, sensitized to antigens in Peyer's patches, migrate via mesenteric lymph nodes and home to intestinal lamina propria. MAdCAM-1 located on endothelial cells regulates this trafficking. Experimentally, parenteral nutrition (PN) decreases GALT cell mass and mucosal immunity when compared with enteral feeding. METHODS: In experiment 1, MAdCAM-1 expression was quantified in 32 mice after 4 days of feeding chow, a complex diet, intragastric (IG)-PN, or PN. In experiment 2, MAdCAM-1 was measured in 102 mice 0, 4, 8, 12, 24, 48, or 72 hours after starting PN and at 0, 4, 8, 12, 24, or 48 hours after reinstituting chow following 5 days of PN. In experiment 3, 56 mice received chow, PN, chow + MECA-367 (anti-MAdCAM-1 mAb), or chow + Isotype control Ab (IsoAb) for 5 days, followed by Peyer's patches, lamina propria, and intraepithelial lymphocyte yield with respiratory and intestinal IgA levels. In experiment 4, 10 days after Pseudomonas immunization, mice received chow + MECA-367 or chow + IsoAb for 4 days followed by 1.2 x 108 Pseudomonas intratracheally. RESULTS: Diet and route affect MAdCAM-1 expression (chow > complex diet > IG-PN > PN). Decreased MAdCAM-1 expression occurred within hours of starting PN in Peyer's patches, but not mesenteric lymph nodes or the intestine, and recovered quickly with enteral refeeding. MAdCAM-1 blockade reduced all GALT populations. Blockade had little effect on IgA levels and partially impaired the late response of established respiratory immunity. CONCLUSIONS: Enteral feeding affects MAdCAM-1 expression. Complete MAdCAM-1 blockade reduces GALT lymphocytes to PN levels, but the chow feeding stimulus preserves IgA and early antibacterial resistance, implying the existence of non-MAdCAM-1 mechanisms to preserve mucosal immunity. PMID- 12724635 TI - Preoperative portal vein embolization for extended hepatectomy. AB - OBJECTIVE: To examine the authors' experience with preoperative ipsilateral portal vein embolization (PVE) and assess its role in extended hepatectomy. SUMMARY BACKGROUND DATA: Extended hepatectomy (five or more liver segments) has been associated with higher complication rates and increased postoperative liver dysfunction than have standard hepatic resections involving lesser volumes. Recently, PVE has been used in patients who have a predicted (postresection) future liver remnant (FLR) volume less than 25% of total liver volume in an attempt to increase the FLR and reduce complications. METHODS: Sixty patients from 1996 to 2002 were reviewed. Thirty-nine patients had PVE preoperatively. Eight patients who had PVE were not resected either due to the discovery of additional unresectable disease after embolization but before surgery (n = 5) or due to unresectable disease at surgery (n = 3). Therefore, 31 patients who had PVE subsequently underwent extended hepatic lobectomy. A comparable cohort of 21 patients who had an extended hepatectomy without PVE were selected on the basis of demographic, tumor, and liver volume characteristics. Patients had colorectal liver metastases (n = 30), hepatocellular carcinoma (n = 15), Klatskin tumors (n = 9), peripheral cholangiocarcinoma (n = 3), and other tumors (n = 3). The 52 resections performed included 42 extended right hepatectomies, 6 extended left hepatectomies, and 4 right hepatectomies extended to include the middle hepatic vein and the caudate lobe but preserving the majority of segment 4. Concomitant vascular reconstruction of either the inferior vena cava or hepatic veins was performed in five patients. RESULTS: There were no differences between PVE and non-PVE groups in terms of tumor number, tumor size, tumor type, surgical margin status, complexity of operation, or perioperative red cell transfusion requirements. The predicted FLR was similar between PVE and non-PVE groups at presentation. After PVE the FLR was higher than in the non-PVE group. No complications were observed after PVE before resection. There was no difference in postoperative mortality, with one death from liver failure in the non-PVE group and no operative mortality in the PVE group. Postoperative peak bilirubin was higher in the non-PVE than the PVE group, as were postoperative fresh-frozen plasma requirements. Liver failure (defined as the development of encephalopathy, ascites requiring sustained diuretics or paracentesis, or coagulopathy unresponsive to vitamin K requiring fresh-frozen plasma after the first 24 hours postresection) was higher in the non-PVE patients than the PVE patients. The hospital stay was longer in the non-PVE than the PVE group. CONCLUSIONS: Preoperative PVE is a safe and effective method of increasing the remnant liver volume before extended hepatectomy. Increasing the remnant liver volume in patients with estimated postresection volumes of less than 25% appears to reduce postoperative liver dysfunction. PMID- 12724636 TI - Champ Lyons: an incomplete life. AB - The life of Champ Lyons, MD, is presented, with emphasis on his tenure as Chairman of the Department of Surgery of the Medical College of Alabama (University of Alabama in Birmingham School of Medicine) from 1950 until 1965. Before becoming chairman Dr. Lyons, as an esteemed microbiologist, participated in the early use of penicillin in wounded servicemen during World War II. Later in his career, he made many contributions to the emerging disciplines of vascular and cardiac surgery. After a brief illness in 1965, Dr. Lyons expired due to a brain tumor. His relatively brief career and his unanticipated sudden demise have lessened the fame of Dr. Lyons to which he is justly entitled. It is the purpose of this presentation to reawaken the surgical community to the importance of this giant of American surgery. PMID- 12724637 TI - Intraperitoneal resuscitation improves intestinal blood flow following hemorrhagic shock. AB - OBJECTIVE: To study the effects of peritoneal resuscitation from hemorrhagic shock. SUMMARY BACKGROUND DATA: Methods for conventional resuscitation (CR) from hemorrhagic shock (HS) often fail to restore adequate intestinal blood flow, and intestinal ischemia has been implicated in the activation of the inflammatory response. There is clinical evidence that intestinal hypoperfusion is a major factor in progressive organ failure following HS. This study presents a novel technique of peritoneal resuscitation (PR) that improves visceral perfusion. METHODS: Male Sprague-Dawley rats were bled to 50% of baseline mean arterial pressure (MAP) and resuscitated with shed blood plus 2 equal volumes of saline (CR). Groups were 1) sham, 2) HS + CR, and 3) HS + CR + PR with a hyperosmolar dextrose-based solution (Delflex 2.5%). Groups 1 and 2 had normal saline PR. In vivo videomicroscopy and Doppler velocimetry were used to assess terminal ileal microvascular blood flow. Endothelial cell function was assessed by the endothelium-dependent vasodilator acetylcholine. RESULTS: Despite restored heart rate and MAP to baseline values, CR animals developed a progressive intestinal vasoconstriction and tissue hypoperfusion compared to baseline flow. PR induced an immediate and sustained vasodilation compared to baseline and a marked increase in average intestinal blood flow during the entire 2-hour post resuscitation period. Endothelial-dependent dilator function was preserved with PR. CONCLUSIONS: Despite the restoration of MAP with blood and saline infusions, progressive vasoconstriction and compromised intestinal blood flow occurs following HS/CR. Hyperosmolar PR during CR maintains intestinal blood flow and endothelial function. This is thought to be a direct effect of hyperosmolar solutions on the visceral microvessels. The addition of PR to a CR protocol prevents the splanchnic ischemia that initiates systemic inflammation. PMID- 12724638 TI - Rapid parathyroid hormone analysis during venous localization. AB - OBJECTIVE: To determine the usefulness of the rapid parathyroid hormone (PTH) assay during venous localization for primary hyperparathyroidism (1 degrees HPTH). SUMMARY BACKGROUND DATA: Remedial exploration for persistent 1 degrees HPTH poses a significant challenge when noninvasive preoperative localization studies are negative. Based on experience with the intraoperative rapid PTH assay, this technique was extrapolated to the interventional radiology suite and generated near real-time data for the interventional radiologist employing on site hormone analysis, with a 12-minute turnaround time from blood sampling to assay result. METHODS: Between November 1997 and July 2002, 446 patients with 1 degrees HPTH were referred for treatment. Of these, 56 (12.5%) represented remedial patients who had each undergone one or more previous cervical explorations. Noninvasive imaging studies were positive for or suggestive of localized disease in 49/56 (87.5%) of these patients, who therefore proceeded directly to surgical exploration. Seven patients with persistent 1 degrees HPTH and negative noninvasive studies underwent selective venous sampling employing a rapid PTH assay in the interventional suite. RESULTS: Venous localization demonstrated an apparent PTH gradient in six of the seven patients. In three, a subtle gradient demonstrated in near real-time prompted additional sampling, which confirmed an unequivocal hormone gradient. In an additional case, the absence of a gradient on initial sampling prompted further sampling, which was positive. All of the patients were explored, and in five of the six patients with a positive PTH gradient, a parathyroid adenoma (mean weight 636 +/- 196 mg) was resected from a location predicted by venous localization. In the sixth patient with a positive gradient, parathyroid tissue was not identified; however, there was a significant fall in the intraoperative PTH values, and immediate postoperative and follow-up laboratory data at 1 month are indicative of a cure. In the one patient with negative localization, abnormal parathyroid tissue could not be located during surgical exploration. CONCLUSIONS: The rapid PTH assay is a major adjunct for obtaining informative venous localization in patients with persistent 1 degrees HPTH. This information is extremely helpful to the surgeon in this challenging group of patients and resulted in a 100% cure rate when a venous gradient was demonstrated. The authors now employ this technique routinely in remedial patients with negative noninvasive imaging studies. PMID- 12724639 TI - Sestamibi scanning and minimally invasive radioguided parathyroidectomy without intraoperative parathyroid hormone measurement. AB - OBJECTIVE: To evaluate the results of a large series of patients undergoing minimally invasive radioguided parathyroidectomy (MIRP) in which routine use of the intraoperative parathyroid hormone assay was not used, and to investigate characteristics between patients who had positive preoperative parathyroid scans versus those with negative scans. SUMMARY BACKGROUND DATA: The technique of parathyroidectomy has traditionally involved bilateral exploration of the neck under general endotracheal anesthesia. Parathyroid imaging using technetium-99m sestamibi (MIBI) has evolved and can localize the adenomas in 80% to 90% of patients. The MIRP technique combines parathyroid scintigraphy with a hand-held gamma detector used intraoperatively to guide the surgeon to the adenoma in patients with positive MIBI scans. Central to this technique or other unilateral approaches is a positive MIBI scan. METHODS: One hundred seventy-three patients with primary hyperparathyroidism operated on by a single surgeon between January 1998 and July 2002 were included. One hundred twelve patients underwent the MIRP procedure and by definition had a positive preoperative parathyroid scan. The technique involved injecting 20 mCi MIBI 1 hour before the surgical procedure in patients who preoperatively had positive MIBI imaging. Patients had the choice of general or MAC anesthesia. Using an incision of less than 4 cm, the dissection to the adenoma was guided by the Navigator 11-mm probe. These 112 patients and 4 additional patients who for various reasons did not have the MIRP procedure yet had positive MIBI scans were compared to 57 patients who had clearly negative MIBI parathyroid imaging. RESULTS: Follow-up data were available for 108 of 112 patients who underwent MIRP. No patients had persistent hypercalcemia. The long term success rate for the MIRP group was 98%. Fifty-two percent of the MIRP procedures were performed using MAC anesthesia. Overall, gland weight and serum PTH were related to the probability of a positive MIBI scan. Multiple logistic regression revealed that females were more likely to exhibit positive scans than were males for any fixed serum PTH level. For females, there was a significant relationship between increasing serum parathyroid hormone and a positive MIBI scan. Conversely, in males, the relationship between scan positivity and serum parathyroid hormone was weaker. CONCLUSIONS: The MIRP technique without routine intraoperative serum parathyroid hormone measurement resulted in an excellent cure rate for primary hyperparathyroidism. As the MIRP technique as well as other techniques for unilateral cervical exploration are predicated on a positive parathyroid scan, the possible effect of gender on the sensitivity of MIBI scintigraphy for the detection of parathyroid adenomas warrants further investigation. PMID- 12724640 TI - Axillary lymph node status, but not tumor size, predicts locoregional recurrence and overall survival after mastectomy for breast cancer. AB - OBJECTIVE: To assess the significance of axillary lymph node status and tumor size for predicting locoregional recurrence (LRR) and overall survival after mastectomy for breast cancer and to discuss the utility of postmastectomy radiation therapy. SUMMARY BACKGROUND DATA: Patients with locally advanced breast cancer require multimodality treatment combining chemotherapy (and/or hormonal therapy), surgery, and radiation. Randomized trials have demonstrated that postmastectomy radiation reduces LRR, but no overall survival benefit has been established. METHODS: Criteria for accrual to the Alabama Breast Cancer Project (1975-1978) were female gender and T2-3 breast cancer with M0 status. Patients underwent a radical or a modified radical mastectomy. Node-positive patients received adjuvant cyclophosphamide, methotrexate, and fluorouracil chemotherapy or adjuvant melphalan. Patients were evaluated for LRR and overall survival based on the number of positive axillary lymph nodes and (in N0 patients) pathologic tumor size. Significance was determined using chi-square analysis. Survival curves were generated using the Kaplan-Meier method and were compared by log-rank analysis. RESULTS: After median follow-up of 15 years, neither type of surgery nor chemotherapy was shown to affect locoregional disease-free or overall survival. LRR rates were higher and overall survival rates were lower in patients with nodal involvement, while tumor size was not shown to significantly affect these rates. CONCLUSIONS: Patients with axillary lymph node metastases may benefit from postmastectomy radiation, but the use of postmastectomy radiation in N0 patients is not supported when it is based on tumor size alone. PMID- 12724641 TI - Increased expression of ceruloplasmin in the retina following photic injury. AB - PURPOSE: Oxidative stress plays a role in the photic injury model of retinal degeneration and in age-related macular degeneration. Our preliminary microarray analysis of retinal gene expression upon photic injury suggested increased expression of ceruloplasmin, a ferroxidase that could reduce retinal oxidative stress. Patients with acerul oplasminemia have retinal degeneration, indicating that ceruloplasmin is necessary for maintenance of retinal health. The purpose of this study was to determine whether retinal ceruloplasmin is upregulated following photo-oxidation, to localize ceruloplasmin protein, and to determine which ceruloplasmin isoform is present in the retina. METHODS: Balb/c mice were exposed to bright white light for seven hours. TUNEL labeling was used to detect photoreceptor apoptosis. At several intervals after the light injury, retinal ceruloplasmin was studied by quantitative PCR, immunohistochemistry, and western analysis. Expression of the secreted and expression of the membrane-anchored glycosyl phosphatidyl inositol (GPI) linked forms of ceruloplasmin were assesed in rat retina using primers specific for each form. Vitreous ceruloplasmin was detected by immunohistochemistry in Balb/c mouse eyes and by western analysis of aspirated vitreous from post-mortem human eyes. RESULTS: Retinal ceruloplasmin mRNA was upregulated eight-fold following photic injury. Ceruloplasmin protein was detected throughout normal retinas by immunohistochemistry, with a specific increase in Muller cell labeling following photic injury. Western analysis confirmed an increase in ceruloplasmin protein following photic injury and revealed eight-fold more ceruloplasmin protein in normal retina than in brain. The mRNAs for both the secreted and GPI linked forms of ceruloplasmin were detected by RT-PCR in the retina. Ceruloplasmin protein was detected by western analysis of normal human vitreous and was increased in mouse vitreous following photic injury. CONCLUSIONS: Ceruloplasmin, a retinal ferroxidase, is upregulated at the mRNA and protein levels upon light damage. The increased protein is primarily in Muller cells. Ceruloplasmin is considerably more abundant in retina than in brain. The retina expresses both the GPI-linked and secreted forms of ceruloplasmin, and since vitreous ceruloplasmin increases following photic injury, some of the retinal ceruloplasmin may be secreted into the vitreous. Ceruloplasmin may protect the retina from oxidative stress by decreasing the amount of ferrous iron available to produce reactive oxygen species. PMID- 12724642 TI - Caspase-3 inhibitor reduces apototic photoreceptor cell death during inherited retinal degeneration in tubby mice. AB - PURPOSE: The tubby mouse, previously suggested as an animal model for the human Usher Syndrome type I, was used in an analysis of pathophysiological processes leading to the inherited retinal degeneration, also shown in Usher syndrome patients. To evaluate pathogenic mechanisms causing retinal degeneration in tubby mice, we examined the time course of apoptotic photoreceptor cell death. Apoptotic pathways were determined by the inhibition of specific caspases in vivo. METHODS: Apoptotic cells were identified during retinal differentiation and degeneration by the TUNEL-method. Apoptotic events were confirmed by DNA laddering. Intravitreal injection of apoptosis inhibitors was applied to reduce apoptotic photoreceptor cell death in tubby mice. RESULTS: During retinal differentiation there is no apparent difference between tubby and wild type mice in apoptotic events. Between post natal day 16 and 23, apoptosis was detected in the outer nuclear layer of tubby mice retinas, but was absent in control mice. The number of TUNEL-labeled photoreceptor cells peaked at post natal day 19. After this peak of apoptosis, the number of apoptotic photoreceptor cells gradually decreased. While a caspase-1 inhibitor did not reduce the number of apoptotic cells, a specific caspase-3 inhibitor caused a significant decrease of apoptotic photoreceptor cells in the tubby mouse retina. CONCLUSIONS: Apoptosis is necessary for appropriate differentiation of the retina of tubby and wild type mice. In the fully developed tubby mouse retina, apoptotic photoreceptor cell death leads to retinal degeneration. Apoptosis in the tubby mouse retina is mediated by specific activation of members of the caspase-3 family. Caspase-3 inhibition drastically reduces photoreceptor cell death in the degenerating tubby mouse retina and may be a potential tool for therapeutic strategies of retinal degeneration in human Usher patients. PMID- 12724643 TI - Genetic heterogeneity of butterfly-shaped pigment dystrophy of the fovea. AB - PURPOSE: Butterfly-shaped macular dystrophy (BSMD) has so far only been associated with mutations in the peripherin/RDS gene. The initial aim of our study was to investigate the peripherin/RDS gene as the causative gene in a family with BSMD. Subsequently the putative involvement of the ROM-1 gene, 4 genes expressed in cone photoreceptors, all known non-syndromic macular, retinal pigment epithelium and choroidal dystrophy loci, all known Leber congenital amaurosis loci and all known non-syndromic congenital and stationary retinal disease loci was examined. METHODS: Thirteen members from the original family with autosomal dominant BSMD were examined. The protein coding exons of the peripherin/RDS gene were screened for mutations by sequence analysis. Linkage analysis was performed using markers flanking the peripherin/RDS gene to rule out the presence of a heterozygous deletion. Likewise, involvement of the ROM-1 gene, four cone genes, 41 non-syndromic retinal disease loci and one syndromic retinal disease locus was investigated. RESULTS: Sequence analysis of the peripherin/RDS gene revealed no mutations. In addition, the BSMD phenotype could not be genetically linked to the peripherin/RDS gene, the ROM-1 gene and the four cone genes nor to any of the 42 retinal disease loci. CONCLUSIONS: This study reveals genetic heterogeneity for BSMD by the identification of a BSMD family, which is not associated with a mutation in the peripherin/RDS gene nor with any other known non-syndromic retinal disease gene. PMID- 12724644 TI - Characterization of RP1L1, a highly polymorphic paralog of the retinitis pigmentosa 1 (RP1) gene. AB - PURPOSE: To determine the full-length sequence of a gene with similarity to RP1 and to screen for mutations in this newly characterized gene, named retinitis pigmentosa 1-like 1(RP1L1). Since mutations in the RP1 gene cause autosomal dominant retinitis pigmentosa, it is possible that mutations in RP1's most sequence similar relative, RP1L1, may also be a cause of inherited retinal degeneration. METHODS: A combination of cDNA clone sequencing, RACE, and database analysis were used to determine the RP1L1 mRNA sequence and its genomic organization. PCR analysis, semi-quantitative RT PCR, and in situ hybridization were used to determine the expression pattern of RP1L1. Single-strand conformational analysis and automated sequencing were used to screen probands from 60 adRP families for potential disease-causing mutations in RP1L1. RESULTS: The human RP1L1 gene is encoded in 4 exons, which span 50 kb on chromosome 8p. The length of the RP1L1 mRNA is large, over 7 kb, but its exact length is variable between individuals due to the presence of several length polymorphisms, including a 48 bp repeat. RP1L1 encodes a protein with a minimal length of 2,400 amino acids and a predicted weight of 252 kDa. Expression of RP1L1 is limited to the retina and appears to be specific to photoreceptors. Mutational analysis of 60 autosomal dominant retinitis pigmentosa probands revealed the presence of 38 sequence substitutions in RP1L1. Over half of these substitutions result in alteration of the RP1L1 protein, but none of these substitutions appear to be pathogenic. CONCLUSIONS: The RP1L1 gene encodes a large, highly polymorphic, retinal-specific protein. No RP1L1 disease-causing mutations were identified in any of the samples tested, making it unlikely that mutations in RP1L1 are a frequent cause of autosomal dominant retinitis pigmentosa. Additional experiments will be needed to determine if mutations in RP1L1 cause other forms of inherited retinal degeneration. PMID- 12724647 TI - Improved detection of inferobasal ischemia during dobutamine echocardiography with doppler tissue imaging. AB - OBJECTIVES: The purpose of this study was quantitative evaluation of the inferobasal segment during dobutamine stress echocardiography using Doppler tissue imaging (DTI). BACKGROUND: Overdiagnosis of myocardial ischemia during dobutamine echocardiography is a common problem. DTI may permit more accurate quantitative diagnosis of ischemia. METHODS: A total of 50 patients with normal contraction of the inferobasal segment at rest were referred for dobutamine stress echocardiography. All underwent coronary angiography. Systolic and diastolic myocardial velocities were measured from apical 2-chamber view at rest and at the peak of dobutamine infusion. RESULTS: Stenosis of the right coronary artery >or= 70% was detected in 11 patients. Conventional stress echocardiography was falsely positive in 10.3% and falsely negative in 27.3%. When DTI was combined with conventional stress echocardiography, sensitivity and specificity was 81.8% and 97.4%, respectively. CONCLUSION: DTI may enhance the diagnosis of inferior ischemia during dobutamine echocardiography and can be added to conventional imaging in the treatment of these patients. PMID- 12724645 TI - Analysis of the rdd locus in chicken: a model for human retinitis pigmentosa. AB - PURPOSE: To identify the locus responsible for the blind mutation rdd (retinal dysplasia and degeneration) in chickens and to further characterise the rdd phenotype. METHODS: The eyes of blind and sighted birds were subjected to ophthalmic, morphometric and histopathological examination to confirm and extend published observations. Electroretinography was used to determine age of onset. Birds were crossed to create pedigrees suitable for genetic mapping. DNA samples were obtained and subjected to a linkage search. RESULTS: Measurement of IOP, axial length, corneal diameter, and eye weight revealed no gross morphological changes in the rdd eye. However, on ophthalmic examination, rdd homozygotes have a sluggish pupillary response, atrophic pecten, and widespread pigmentary disturbance that becomes more pronounced with age. Older birds also have posterior subcapsular cataracts. At three weeks of age, homozygotes have a flat ERG indicating severe loss of visual function. Pathological examination shows thinning of the RPE, ONL, photoreceptors and INL, and attenuation of the ganglion cell layer. From 77 classified backcross progeny, 39 birds were blind and 38 sighted. The rdd mutation was shown to be sex-linked and not autosomal as previously described. Linkage analysis mapped the rdd locus to a small region of the chicken Z chromosome with homologies to human chromosomes 5q and 9p. CONCLUSIONS: Ophthalmic, histopathologic, and electrophysiological observations suggest rdd is similar to human recessive retinitis pigmentosa. Linkage mapping places rdd in a region homologous to human chromosomes 9p and 5q. Candidate disease genes or loci include PDE6A, WGN1, and USH2C. This is the first use of genetic mapping in a chicken model of human disease. PMID- 12724646 TI - Human corneal stem cells display functional neuronal properties. AB - PURPOSE: Human corneal limbal stem cells mature and repopulate the superficial layers of the cornea throughout life. In this study, we tested the hypothesis that human corneal stem cells, derived from neural ectoderm, can exhibit functional neuronal properties. METHODS: Human corneal limbal tissue (donor age 6 weeks to 92 years) was obtained from Upstate New York Transplant Services. Tissues were grown as explants on coverslips in DMEM with 10% calf serum. After 7 14 days in vitro, tissues on coverslips were double-immunostained for the stem cell marker, p63, along with nestin and neurotransmitter receptors GABA, dopamine, serotonin, glycine or acetylcholine. We also carried out whole cell current clamp and voltage clamp recordings on corneal stem cell cultures in order to determine their functional neurophysiological properties. RESULTS: Co localization of p63 with nestin, GABA receptor, glycine receptor, and serotonin receptor immunoreactivity was seen in a small number of cells in the corneal stem cell cultures. The resting potential of corneal stem cells was relatively low, approximately -13+/-8 mV (n=13; range -6 mV to -40 mV) measured in current clamp. No action potentials or voltage sensitive Na+ and K+ currents were detected. However, in a small number of cells, kainic acid (0.5 mM), a non-NMDA glutamate receptor agonist, and GABA induced a small inward current. Glutamate receptor antagonist, CNQX, and GABA receptor antagonist, bicuculline and CGP-35348 blocked the agonist response. CONCLUSIONS: A subpopulation of human corneal stem cells exhibit neuronal properties in vitro, as evidenced by immunoreactivity to nestin, GABA receptor, glycine receptor, and serotonin receptor, as well as functional neurophysiological responses to GABA and kainic acid. Human corneal stem cells may represent a potential source of non-embryonic, autologous, surgically accessible graft material with neuronal potential. PMID- 12724648 TI - Temporal changes and histologic relation of postsystolic thickening in an animal model of acute ischemia and reperfusion. AB - BACKGROUND: We investigated the incidence and temporal changes of postsystolic thickening (PST) during myocardial ischemia and reperfusion to determine factors associated with PST. METHODS: The inferoposterior wall infarction was induced by coronary artery ligation (L) in 22 male cats. Epicardial echocardiographic examination including 2-dimensional, M-mode, and pulsed wave myocardial Doppler imaging of inferoposterior wall was performed at short-axis view before L, and regularly repeated during L until PST was not detected by myocardial Doppler image. Reperfusion (R) was done immediately after PST disappeared, and epicardial echocardiographic examination was repeated for 1 hour. Transmural extent of myocardial necrosis was morphometrically analyzed with triphenyltetrazolium chloride staining. RESULTS: PST measured by myocardial Doppler image was well correlated with that by M-mode (r = 0.63, P <.001). Immediately after L, all 22 cats showed akinesia in 2-dimensional imaging; among them PST was present in 13 cats (59%) and absent in the other 9 cats. The extent of ischemia by perimetric measurement with 2-dimensional imaging was smaller in cats with PST compared with those without (32 +/- 10% vs 47 +/- 5%, P <.001). In 13 cats with PST, the occlusion time until disappearance of PST (90-270 minutes, 160 +/- 70) and transmural extent of myocardial necrosis (0%-72%, 22 +/- 24) were quite variable. After R, PST promptly reappeared in all 13 cats. PST persisted until 1 hour after R in 5 cats, whereas it disappeared in 8 cats as systolic thickening became predominant; occlusion time was significantly longer (220 +/- 40 vs 120 +/- 40 minutes, P <.005) and transmural extent of myocardial necrosis was larger (43 +/- 17% vs 8 +/- 15%, P <.005) in cats with persistent PST until 1 hour after R. CONCLUSION: In acute ischemia and R, we observed variable patterns of PST genesis and maintenance, and different factors were related with these phenomena in each stage. PST could be used not only as a marker of acute ischemia but also as a marker of successful myocardial R, and further study is necessary to test whether PST represents different status of myocardial mechanics according to the extent of myocardial ischemia and necrosis in the clinical setting. PMID- 12724649 TI - Incidence and characteristics of segmental postsystolic longitudinal shortening in normal, acutely ischemic, and scarred myocardium. AB - OBJECTIVE: Myocardial longitudinal shortening after aortic valve closure (postsystolic shortening [PSS]) is considered a marker of pathology with diagnostic potential. However, PSS can also occur in healthy subjects. We, therefore, investigated the occurrence and characteristics of PSS in control subjects and patients, and how to distinguish normality from disease. METHODS: In 20 young control subjects, 10 older control subjects, 30 patients with acute myocardial infarction (acute ischemia), and 10 patients with postischemic myocardial scar, longitudinal myocardial deformation was measured with Doppler tissue strain rate (SR) imaging. Segmental SR and strain were visually and quantitatively analyzed and compared. RESULTS: In young control subjects, PSS was found in 98 of 313 segments (31%) and showed gaussian distribution (median 1.3%). During ejection time, median peak SR was -1.4 s(-1) and median strain -16.6%. In older control subjects, parameters differed only slightly. In acutely ischemic and scarred myocardium, both systolic strain and SR were significantly reduced or inverted. In disease, PSS occurred significantly more often (78% and 79%, respectively), was significantly higher in magnitude, and its peak occurred later than in young and older control subjects. CONCLUSION: PSS is a normal finding in healthy subjects occurring in approximately one-third of myocardial segments and, thus, is not always a marker of disease. Our data indicate that pathologic PSS can be detected by coexisting reduction in systolic strain and, second, by exceeding a postsystolic strain magnitude cutoff. PMID- 12724650 TI - An evaluation of the use of new Doppler methods for detecting longitudinal function abnormalities in a pacing-induced heart failure model. AB - BACKGROUND: Doppler tissue echocardiography and color M-mode Doppler flow propagation velocity have proven useful in evaluating cross-sections of patients with left ventricular (LV) dysfunction, but experience with serial changes is limited. PURPOSE AND METHODS: We tested their use by evaluating the temporal changes of LV function in a pacing-induced congestive heart failure model. Rapid ventricular pacing was initiated and maintained in 20 dogs for 4 weeks. Echocardiography was performed at baseline and weekly during brief pacing cessation. RESULTS: With rapid pacing, LV volume significantly increased and ejection fraction (57%-28%), stroke volume (37-18 mL), and mitral annulus systolic velocity (16.1-6.6 cm/s) by Doppler tissue echocardiography significantly decreased, with ejection fraction and mitral annulus systolic velocity closely correlated (r = 0.706, P <.0001). In contrast to the mitral inflow velocities, mitral annulus early diastolic velocity decreased steadily (12.3-7.3 cm/s) resulting in a dramatic decrease in mitral annulus early/late (1.22-0.57) diastolic velocity with no tendency toward pseudonormalization. The color M-mode Doppler flow propagation velocity also showed significant steady decrease (57-24 cm/s) throughout the pacing period. Multiple regression analysis chose mitral annulus systolic velocity (r = 0.895, P <.0001) and propagation velocity (r = 0.782, P <.0001) for the most important factor predicting LV systolic and diastolic function, respectively. CONCLUSIONS: Doppler tissue echocardiography and color M-mode Doppler flow could evaluate the serial deterioration in LV dysfunction throughout the pacing period. These were more useful in quantifying progressive LV dysfunction than conventional ehocardiographic techniques, and were probably relatively independent of preload. These techniques could be suitable for longitudinal evaluation in addition to the cross-sectional study. PMID- 12724651 TI - Differential changes in regional right ventricular function before and after a bilateral lung transplantation: an ultrasonic strain and strain rate study. AB - The evaluation of regional right ventricular function by ultrasound remains a challenge. This case report demonstrates the potential value of the new cardiac deformation indices, strain and strain rate imaging, in determining the differing regional abnormalities in longitudinal right ventricular function before and after bilateral lung transplantation. These indices were measured in a patient with severe right ventricular dysfunction as a result of primary pulmonary hypertension. PMID- 12724652 TI - Usefulness of the mitral/aortic flow velocity integral ratio as a screening method to identify patients with hemodynamically significant mitral regurgitation. AB - OBJECTIVE: We sought to test the value of a simple Doppler index, the mitral/aortic flow velocity integral ratio (MAVIR), as a screening method to identify patients with hemodynamically significant mitral regurgitation (MR). METHODS: Included in the study were 91 patients (mean age 61 +/- 14 years; 54% men) with echocardiographically diagnosed MR. The cause was organic in 23 patients and ischemic/functional in 68. MR degree was assessed by 2 quantitative echocardiographic measures: the regurgitant fraction and the regurgitant orifice area. RESULTS: A good correlation was found between MAVIR and both regurgitant fraction (r = 0.75) and orifice (r = 0.60). When we divided patients into 3 groups on the basis of the regurgitant fraction, used as reference standard, MAVIR significantly increased in proportion to MR severity (0.7 +/- 0.1 for mild MR vs 1.1 +/- 0.2 for moderate and 1.4 +/- 0.3 for severe regurgitation; P <.0001). A ratio > 1 identified 28 of 30 patients with severe MR (regurgitant fraction > 60%), whereas all patients with mild MR (regurgitant fraction < 40%) had a ratio < 1. Using a regurgitant orifice >or= 40 mm(2) as threshold for severe MR, a significant difference in MAVIR was also present among patients who had severe MR compared with those having mild and moderate regurgitation (P <.0001). CONCLUSIONS: These findings suggest that MAVIR is a sensitive index, potentially widely applicable in clinical practice as a screening parameter for identifying patients with hemodynamically significant MR. PMID- 12724653 TI - Correlation of the Tei index with invasive measurements of ventricular function in a porcine model. AB - BACKGROUND: The Doppler myocardial performance (Tei) index has been reported to be clinically useful in assessing left ventricular systolic and diastolic function in both adults and children. However, there are limited data to compare the Tei index with invasive measurements of ventricular function. We used a porcine model to directly correlate the Tei index with invasive indices of systolic and diastolic function. METHODS: Pressure volume loops were obtained from 10 pigs (32-45 kg). A micromanometer and a conductance catheter were placed in the left ventricle to record pressure and volume, respectively. A flow probe was placed around the ascending aorta to record cardiac output. Baseline pressure volume loops were generated during preload reduction through caval occlusion. Epicardial echocardiograms were performed just before the caval occlusion. Invasive indices including preload recruitable stroke work, ventricular stiffness constant, and cardiac output were assessed, as were noninvasive echocardiographic indices including Tei index and ejection fraction. An ischemic insult, ventricular fibrillation, was induced to alter ventricular function. After cardioversion and 40 minutes of reperfusion, echocardiographic and invasive measurements were repeated. RESULTS: There was a statistically significant inverse relationship between the percent change in Tei and the percent change in preload recruitable stroke work after ventricular fibrillation (r = -0.70, P =.02), although the correlation between the actual values of Tei and preload recruitable stroke work were not statistically significant. There was a statistically significant inverse relationship between the percent change in Tei and the percent change in cardiac output (r = -0.65, P =.03). There was a direct correlation between the value of Tei and the ventricular stiffness constant at baseline (r = 0.63, P <.05). As anticipated, the value of Tei was inversely related to ejection fraction by epicardial echocardiogram at baseline (r = -0.85, P <.001). The percent change in Tei was inversely related to the percent change in ejection fraction as well (r = -0.69, P <.05). CONCLUSIONS: This animal model is one of the first studies to demonstrate a direct correlation between the Tei index and systolic and diastolic invasive measurements of ventricular function. This supports the clinical use of this index as a measure of global ventricular function. PMID- 12724654 TI - Single-plane and biplane echocardiography: use of targeted scan planes improves the estimates of left ventricular volume and shape for analysis of postinfarction remodeling. AB - OBJECTIVE: Acute myocardial infarction and subsequent left ventricular (LV) remodeling induce complex geometric changes quantifiable by 3-dimensional (3D) echocardiography. Our objective was to determine accurate 2-dimensional echocardiographic techniques for analysis of diastolic and systolic LV volume and shape in remodeled hearts. METHODS: We obtained 3D apical scans from 16 patients at the acute stage, and at 1 and 6 months after acute myocardial infarction. LV volumes were calculated by 7 methods: (1) Teichholz; (2) Teichholz including the infarcted area; (3) single-plane area-length (AL) using a 2-chamber (2CH) view; (4) single-plane AL using a 4-chamber (4CH) view; (5) single-plane AL using a view including the infarcted area; (6) biplane AL using 2CH and 4CH views; and (7) biplane AL using a view including the infarction region and corresponding orthogonal view (method ALBMIO). LV shape was assessed by 5 methods: (1) a 2CH view; (2) a 4CH view; (3) a single-plane view including the infarcted area; (4) biplane (2CH and 4CH) views; and (5) biplane views including the infarction region and corresponding orthogonal view (method BMIO). RESULTS: For end diastolic and end-systolic LV volume assessments, all 7 methods correlated with the 3D reference, but method ALBMIO performed best (end-diastolic: r = 0.931, bias = 17.4 mL; end-systolic: r = 0.946, bias = 11.2 mL). For LV shape assessments, method BMIO showed the smallest difference from the 3D reference. CONCLUSIONS: With 2-dimensional echocardiographic techniques, quantitative analysis of LV volume and shape is most accurate when a component scan plane is targeted through the infarcted myocardial region. PMID- 12724656 TI - Feasibility of noninvasive transthoracic echocardiography/Doppler measurement of coronary flow reserve in left anterior descending coronary artery in patients with acute coronary syndrome: a new technique tested in clinical practice. AB - OBJECTIVE: The aim of this study was to test the feasibility and accuracy of transthoracic Doppler echocardiography measurement of coronary flow reserve (CFR) in the left anterior descending coronary artery (LAD) territory in the clinical setting of the acute coronary syndrome. METHODS: Transthoracic Doppler echocardiography measurements of CFR were made in 42 consecutive patients in the distal LAD before and during adenosine infusion. The results were validated by coronary angiography. A normal CFR was predefined as a more than 2-fold increase of flow velocity during adenosine infusion. RESULTS: We were able to detect significant stenosis in the LAD territory with 92% sensitivity and 82% specificity if we considered a stenosis >or= 50% to be significant. Defining a stenosis of >or= 70% as significant increased the sensitivity and the negative predictive value to 100%, with a specificity of 70%. CONCLUSION: Measuring CFR using transthoracic Doppler echocardiography is noninvasive, feasible, accurate, and relatively inexpensive. The excellent negative-predictive value of this technique makes it a useful tool for identifying patients who can avoid repeated angiography as a result of suspected subacute LAD restenosis after percutaneous coronary intervention. PMID- 12724655 TI - Simultaneous assessment of wall motion and coronary flow velocity in the left anterior descending coronary artery during dipyridamole stress echocardiography. AB - OBJECTIVES: This study was designed to assess the feasibility and clinical meaning of simultaneous assessment of wall motion and coronary flow velocity (CFV) reserve in the left anterior descending coronary artery (LAD) by transthoracic approach for detecting LAD stenosis during dipyridamole stress echocardiography. BACKGROUND: Coronary flow reserve plays an important role, which can be evaluated by transthoracic Doppler echocardiography during vasodilator stress. METHODS: Dipyridamole stress test was performed in 110 patients with known or suspected coronary artery disease. CFV in the distal LAD was obtained at baseline and after dipyridamole infusion, and wall motion was also assessed up to the administration of atropine, if required. All patients underwent quantitative coronary angiography within 2 days of the stress test, and significant LAD stenosis was defined as > 50% stenosis. RESULTS: The success rate of both measurements was 92%. CFV reserve < 2 had a higher sensitivity (94% vs 72%, P <.01) and a lower specificity (65% vs 95%, P <.01) than wall-motion assessment for detecting significant LAD stenosis, and diagnostic accuracy between 2 methods was comparable (81% vs 82%). A total of 69 patients (73%) showed concordant results of the 2 methods, and diagnostic accuracy for detecting significant LAD stenosis was high (94%) in this subset of patients. CONCLUSIONS: The simultaneous assessment of CFV and wall motion was feasible in the majority of cases during dipyridamole stress echocardiography. Although diagnostic accuracy between the 2 tests was comparable, concordant results of the 2 methods provided accurate diagnosis in detecting significant LAD stenosis. PMID- 12724657 TI - Coronary blood flow reserve response to left anterior descending coronary artery stenting and its value in predicting coronary restenosis. AB - BACKGROUND: Heterogeneous results of coronary flow velocity reserve (CFVR) to percutaneous procedures have been reported and the impact of transesophageal echocardiographic evaluation of CFVR in predicting restenosis has not been completely established. METHODS AND RESULTS: We studied 20 control volunteers and 51 patients with left anterior descending coronary artery stenosis to determine the CFVR response to left anterior descending coronary artery stenting, the clinical markers of persistent CFVR impairment, and its value in predicting restenosis. Prestent CFVR was lower in the stenosis group than in control volunteers (1.89 +/- 0.66 vs 3.82 +/- 1.15; P <.001). Although there was a significant increase of CFVR after stenting (2.58 +/- 0.76; P <.001 vs prestent), it remained depressed in 53% of patients and was independently related to multivessel disease (odds ratio, 0.14; 95% confidence interval 0.03-0.55; P =.005), age (odds ratio, 1.07; 95% confidence interval 0.99-1.15; P =.056), and prestent CFVR (odds ratio, 3.78; 95% confidence interval 0.99-14.42; P =.051). CFVR measured both before and early after stenting did not differ between patients with and without restenosis. CONCLUSIONS: CFVR impairment occurs in a large proportion of patients despite successful stenting and appears to be consequent of the extent of atherosclerotic coronary disease. Periprocedural CFVR conferred no predictive value for subsequent intrastent restenosis. PMID- 12724658 TI - Echocardiographic guidance for transcatheter coil occlusion of patent ductus arteriosus in the catheterization laboratory. AB - The objective of this study was to investigate the feasibility of transthoracic echocardiography as a guide to monitor the completeness of patent ductus arteriosus (PDA) occlusion immediately after coil implantation. In all, 52 patients who underwent Gianturco coil implantation for PDA occlusion were evaluated by transthoracic echocardiography within 15 minutes after the first coil implantation in the catheterization laboratory. According to Doppler echocardiographic findings, these patients were classified into 3 groups: group 1, complete occlusion without residual shunt; group 2, a residual PDA diameter < 1 mm and no continuous waveforms detected; and group 3, a residual PDA diameter >or= 1 mm or continuous waveforms. There were 34 patients in group 1, 10 patients in group 2, and 8 patients in group 3. In a 12-month follow-up the complete occlusion rate was 100%, 90%, and 87.5% in groups 1, 2, and 3, respectively. We conclude that immediate Doppler echocardiography is useful in assessing the status of residual PDA just after coil implantation. Residual shunt < 1 mm and lack of continuous waveform on Doppler scan indicate sufficient ductal closure of PDA. PMID- 12724659 TI - Hand-carried ultrasound-guided pericardiocentesis and thoracentesis. AB - To date, ultrasound-guided centeses have required the use of large, diagnostic ultrasound equipment that can be cumbersome and not readily available. In this study, we sought to evaluate the use of portable, hand-carried ultrasound devices in guidance of pericardiocentesis and thoracentesis. Hand-carried ultrasound was used to define location of the maximal extent of the effusion and its proximity to the skin surface, confirm needle entry and site, and to evaluate postprocedural result. A total of 12 patients who were stable and hospitalized were prospectively enrolled, 9 for pericardiocentesis and 3 for thoracentesis. We concluded that under the direction of a suitably trained physician, hand-carried ultrasound provided images suitable for guidance of a pericardiocentesis or thoracentesis preprocedurally, intraprocedurally, and postprocedurally. PMID- 12724660 TI - Determination of the optimal region for interaliasing distance measurement for flow regurgitant rate calculation: a fluid simulation study. AB - BACKGROUND: Color Doppler imaging of the convergent region is promising for quantifying valvular regurgitation. Nevertheless, proximal isovelocity surface area method has limitations. We sought to determine the optimal localization to measure the most precise flow rate using a new approach: the interaliasing distance. METHODS: A finite volume-based program was used to perform simulations in unsteady flow conditions. Different instantaneous flow rates, leaflet angles, and orifice sizes were tested reproducing physiologic conditions. Relative difference between actual and interaliasing distance flow rate was calculated for each configuration. RESULTS: The relationship between the relative error and the aliasing velocity location was described by a third-order polynomial equation. The magnitude of relative error is a function of the flow rate, orifice size, and leaflet angle. CONCLUSION: The optimal distance from the orifice to measure the interaliasing distance was when the closer aliasing was between 4 and 8 mm from the orifice. PMID- 12724661 TI - Multiple aortic valve papillary fibroelastoma: an unusual presentation of a rare tumor. AB - Cardiac papillary fibroelastoma is a rare cardiac tumor and occurs mainly on cardiac valves. The incidence of multiple lesions is exceedingly rare and is about 7% of all reported cases. Careful echocardiographic evaluation of the patient before operation is of high importance as the there is no recurrence after surgical excision. We present the first case of a patient with multiple aortic valve papillary fibroelastomas diagnosed before operation. The patient underwent surgical removal of the 3 masses that were confirmed as cardiac papillary fibroelastomas by pathologic examination. There was no evidence of aortic insufficiency after operation. PMID- 12724662 TI - Transesophageal echocardiographic identification of an aortic root to right atrial fistula in a patient with acute streptococcal aortic valve bacterial endocarditis. AB - We describe the case of a patient with acute aortic valve endocarditis complicated by aortic insufficiency, perivalvular abscess, and aortic root to right atrial fistula caused by Streptococcus pneumoniae bacterial infective endocarditis. Although typically of low sensitivity for the diagnosis of myocardial abscess and similar complications of infective endocarditis, transthoracic echocardiogram successfully identified this relatively uncommon complication that was later confirmed by transesophageal echocardiography and at time of operation. PMID- 12724663 TI - Moritz Katzenstein: the father of meniscus repair surgery. PMID- 12724664 TI - A giant ganglion cyst that developed in the infrapatellar fat and partly extended into the knee joint. AB - Ganglion cysts of the knee joint usually present near the lateral meniscus. Their size varies, and they may be encased in single or multiple capsules. Few case reports on ganglion cysts of the knee joint have been reported. We report a giant ganglion cyst that developed in the patellar fat and partly extended into the joint. The ganglion cyst was exposed in the meniscosynovial junction and was electrically evaporated at the anterior horn. No recurrence has occurred to date, and the patient has returned to his previous level of activity. PMID- 12724665 TI - Thermal capsulorraphy of bilateral glenohumeral joints in a pediatric patient with Ehlers-Danlos syndrome. AB - Ehlers-Danlos syndrome (EDS) is a heterogeneous collection of inherited connective tissue disorders characterized by hypermobility of the joints and hyperextensibility and fragility of the skin. For many patients, the hypermobile joints become problematic. To date, the mainstay of surgical treatment for EDS related joint laxity has been open surgical capsulorraphy, which, although usually effective, confers significant morbidity to the patient. We present the case of a 9-year-old girl diagnosed with a variant of EDS and severely disabled from multidirectional instability of her shoulders and recurrent dislocations of her hips. After 1 year of nonoperative treatment (physical therapy, bracing, and activity restriction) failed, we performed a sequential arthroscopic thermal capsulorraphy of both shoulders. At a 2-year follow-up, the patient has no instability in the left shoulder and only occasional subluxations of the contralateral shoulder. We believe that thermal capsulorraphy is a viable addition to the shoulder surgeon's armamentarium in treating multidirectional instability in children with EDS. PMID- 12724666 TI - Partial deficiency of the lateral meniscus. AB - We report an abnormally shaped lateral meniscus. As far as we know, the present case is the first description of partial deficiency of the lateral meniscus in the literature. Although it is a rare abnormality, arthroscopic surgeons should be familiar with this entity for the differential diagnosis of bucket-handle meniscal tears, not only for magnetic resonance imaging, but also for arthroscopic evaluation. This entity could also be an aggravating factor for instability symptoms of partial or total anterior cruciate ligament-deficient knees. PMID- 12724667 TI - Posterior tibial subluxation and short-term arthritis resulting from failed posterior cruciate ligament reconstruction. AB - Posterior tibial subluxations because of combined or isolated posterior cruciate ligament (PCL) injuries require detailed evaluation. PCL reconstructions are difficult procedures because of the low rate of such injuries and the complex anatomy of the ligament. We report on 2 cases of failed PCL reconstruction because of malpositioned femoral tunnels. These 2 cases support the existing biomechanical evidence that the correct placement of the tunnels, especially in the femur, is a major factor in defining the outcome. It seems that the drilling of the tunnels, especially in the femur, during PCL reconstruction must be performed with accuracy and always be evaluated in cases of graft failure. In addition, failed PCL reconstructions are usually accompanied by a short-term excessive arthritis that results in poor functional outcome. PMID- 12724668 TI - Technical tips in orthopaedics: meniscal repair with use of an in situ fibrin clot. AB - The importance of meniscal preservation has become increasingly clear, and many authors advocate repair of the meniscus whenever possible, forgoing patient age and tear location. In accordance with the common goal of preserving meniscal tissue, we describe a simple technique to augment meniscal repairs with the use of an in situ fibrin clot. The site is prepared accordingly: the synovium directly above the tear site is abraded with a shaver, rasp, or small intra articular banana blade. Next, the water inflow cannula is closed, and the outflow cannula is opened to vacuum suctioning, allowing collapse of the distended joint. Under these circumstances, negative intra-articular pressure is produced in the knee joint. This condition is maintained for a period of 1 to 2 minutes and induces bleeding from the abraded synovial site. The knee in a dependent position causes the blood to run down the synovial wall and into the meniscal cleft, pooling there and forming a clot adherent to the edges of the separated meniscal tear. This procedure has been found to be a simple and minimally invasive mechanism for clot placement. The exposed collagen of the meniscal tear provides an ideal surface for a relatively tenacious clot attachment. It provides the healing factors reported to induce successful meniscal healing. PMID- 12724670 TI - Identification of the saphenous nerve at arthroscopy. AB - Injury to the saphenous nerve can be a troublesome complication of arthroscopic procedures to the posterior half of the medial meniscus. In this article, we outline a technique for accurately identifying saphenous nerve position using transillumination of the saphenous vein. PMID- 12724669 TI - Arthroscopically assisted reconstruction and percutaneous screw fixation of a Pilon tibial fracture. AB - We report a case of closed distal tibial fracture (AO 43C3), treated successfully with arthroscopically assisted minimally invasive reduction and percutaneous screw fixation. Techniques and postoperative treatment are described. PMID- 12724671 TI - Surgical technique and rationale for medial patellofemoral ligament reconstruction for recurrent patellar dislocation. AB - This article describes a medial patellofemoral ligament (MPFL) reconstruction procedure using an artificial ligament for recurrent patellar dislocation. The MPFL has been shown biomechanically to be the primary restraint among the medial patellar stabilizers. Although various predisposing factors are involved in lateral patellar dislocation, we believe that the MPFL, as a primary restraint, should undergo primary reconstruction for patellar dislocation. The results of MPFL reconstruction using a mesh-type artificial ligament and medial retinaculum slip coverage for recurrent patellar dislocation were as good as we had expected. In this article, we describe the detailed surgical technique and its rationale. We believe technique has also wide applications in MPFL reconstruction using autogenous tendons. PMID- 12724672 TI - Arthroscopic osteochondral grafting in fissuring osteochondritis dissecans of the knee. AB - We encountered a rare case of osteochondritis dissecans in a 13-year-old girl, in which magnetic resonance imaging appeared to show migration of the joint cartilage into the bone marrow of the femoral condyle, with fissuring visible on the joint cartilage. Arthroscopy showed fissuring, 5 x 20 mm, in the right lateral femoral condyle, and the articular cartilage in the center of the lesion had fibrillation and was depressed about 5 mm. We performed arthroscopic osteochondral grafting to the lesion. The microscopic findings showed that the lesion had fibrillated bone marrow and that the cartilage existed only in the superficial layer of the lesion. Twenty-seven months after surgery, the patient was able to participate in sports and had no knee pain. PMID- 12724673 TI - Anterior cruciate ligament reconstruction: allograft versus autograft. AB - PURPOSE: This study was performed to compare the minimal 2-year outcome of anterior cruciate ligament (ACL) reconstruction using bone-patellar tendon-bone (BPTB) allografts versus autografts, both augmented with an iliotibial band tenodesis. TYPE OF STUDY: Retrospective review. METHODS: Forty-six of 52 BPTB ACL reconstructions using allografts and 33 of 37 BPTB ACL reconstructions using autografts were followed up at a mean of 2.75 and 3.36 years, respectively. All patients had an iliotibial band tenodesis. Evaluations included the Lysholm II scale, a questionnaire, physical examination findings, and KT-1000 arthrometry. RESULTS: No statistically significant differences were seen between groups in Lysholm II scores or in any subjective category. Most patients (91% allograft; 97% autograft) had good to excellent Lysholm II scores. Sixty-five percent of allograft patients and 73% of autograft patients returned to their preinjury activity level. More allograft patients complained of retropatellar pain (16% v 9% for autograft patients). Fifty-three percent of allograft patients versus 23% of autograft patients had a flexion deficit of 5 degrees or more when compared with the normal contralateral side. When comparing KT-1000 side-to-side differences, we found no significant differences between groups. Ninety-one percent of both groups had maximum side-to-side differences less than 5 mm. Three allograft patients (6.5%) had traumatic ruptures at 12, 19, and 43 months postoperatively versus none in the autograft group. All three allograft patients who sustained postoperative traumatic ruptures had received fresh frozen, nonirradiated allografts. CONCLUSIONS: Results of ACL reconstruction using allografts or autografts augmented with an iliotibial band tenodesis were comparable. The BPTB autograft should remain the gold standard, although the BPTB allograft in ACL reconstruction is a reasonable alternative. PMID- 12724674 TI - Results of rasping of meniscal tears with and without anterior cruciate ligament injury as evaluated by second-look arthroscopy. AB - PURPOSE: Meniscal rasping without suturing has been experimentally shown to stimulate vascular induction in tears in the avascular zone of menisci, resulting in meniscal healing. The goals of this study were to arthroscopically assess the results of meniscal rasping and analyze the factors affecting meniscal healing. TYPE OF STUDY: Retrospective cohort study. METHODS: Forty-eight torn menisci in 47 patients (age range, 14-47 years; average, 24 years) treated arthroscopically with the meniscal rasping technique were evaluated by second-look arthroscopy. The interval between the injury and the time of surgery ranged from 3 weeks to 13 years. There were 35 lateral and 13 medial meniscal tears associated with 44 anterior cruciate ligament injuries; 28 of the menisci had a full-thickness longitudinal tear and the other 20 had a partial-thickness tear. The length of the tears ranged from 10 to 33 mm (mean, 14.4 mm). The distance from the capsule to the tear ranged from 1 to 9 mm (mean, 5.0 mm). RESULTS: Thirty-four menisci (71%) healed completely (without a marked visible unhealed area), 10 (21%) healed incompletely, and 4 (8%) showed no evidence of healing. There were no relationships between outcome and age, gender, injured side, or time from injury and rasping. Both the distance from the capsule to the tear and the length of the tear were longer in the unhealed menisci. Stable tears had a high healing rate after meniscal rasping. CONCLUSIONS: Meniscal rasping without suturing is an easy procedure to perform and seems to be a reliable way to repair longitudinal tears in the avascular region of the meniscus, although the healing potential of the procedure is affected by the distance from the capsule to the tear site and the length and the stability of the tear. PMID- 12724675 TI - The incidence and significance of femoral tunnel widening after quadrupled hamstring anterior cruciate ligament reconstruction using femoral cross pin fixation. AB - PURPOSE: The purpose of this study was to evaluate the amount of femoral tunnel widening that occurred after anterior cruciate ligament reconstruction using quadrupled hamstring autografts and to determine the clinical significance of any such tunnel enlargement. TYPE OF STUDY: Retrospective clinical analysis. METHODS: Twenty-nine patients who had undergone reconstruction of a torn anterior cruciate ligament with quadrupled hamstring autograft and cross pin femoral fixation were evaluated to determine the incidence and significance of postoperative femoral tunnel widening. A single surgeon performed all procedures, and average follow-up was 18.4 months (range, 12 to 31.5 months). All patients underwent flexion posteroanterior and lateral radiographs, an examination for determination of an International Knee Documentation Committee (IKDC) rating, had KT-1000 data collected, and completed Lysholm and Knee Outcome Survey functional questionnaires. Femoral tunnels were clearly seen in 27 patients. The tunnel diameters were measured at the opening of the tunnel, at the widest part of the tunnel, and just proximal to the cross pin. The amount of tunnel widening for each patient was then compared with the individual's KT-1000 data, IKDC rating, and Lysholm and knee outcome survey scores to assess correlation. RESULTS: Four different tunnel morphologies were noted, with the linear type being the most common. The widening at the greatest tunnel diameter was 65.5% on average. Side to-side KT-1000 differences averaged 1.04 mm at 30 lb, and 1.10 mm at manual maximum. Eleven patients had IKDC overall ratings of normal, 13 had ratings that were nearly normal, and 2 had abnormal. Average Lysholm and knee outcome survey scores were 92.6 and 93.9, respectively. A significant correlation was found only between F2 and F3 widening with Lysholm scores. However, the significance was eliminated with removal of 2 outliers. CONCLUSIONS: The exact etiology of postoperative anterior cruciate ligament tunnel widening remains unknown. The present study reveals that significant tunnel widening occurs with quadrupled hamstring autografts and femoral cross pin fixation. However, the widening does not appear to have a significant effect on postoperative ligament laxity or functional knee scores, at least in the short term. PMID- 12724676 TI - Outcomes of microfracture for traumatic chondral defects of the knee: average 11 year follow-up. AB - PURPOSE: In this study, we measured functional outcomes of patients treated arthroscopically with microfracture for full-thickness traumatic defects of the knee. TYPE OF STUDY: A case series of patients with 7 to 17 years' follow-up. METHODS: Between 1981 and 1991, a total of 72 patients (75 knees) met the following inclusion criteria: (1) traumatic full-thickness chondral defect, (2) no meniscus or ligament injury, and (3) age 45 years and younger (range, 13 to 45 years). Seventy-one knees (95%) were available for final follow-up (range, 7 to 17 years). All patients completed self-administered questionnaires preoperatively and postoperatively. RESULTS: The following results were significant at the P <.05 level. Significant improvement was recorded for both Lysholm (scale 1 to 100; preoperative, 59; final follow-up, 89) and Tegner (1 to 10; preoperative, 3; final follow-up, 6) scores. At final follow-up, the SF-36 and WOMAC scores showed good to excellent results. At 7 years after surgery, 80% of the patients rated themselves as "improved." Multivariate analysis revealed that age was a predictor of functional improvement. CONCLUSIONS: Over the 7- to 17-year follow-up period (average, 11.3 years), patients 45 years and younger who underwent the microfracture procedure for full-thickness chondral defects, without associated meniscus or ligament pathology, showed statistically significant improvement in function and indicated that they had less pain. PMID- 12724677 TI - The effect of outlet fixation on tunnel widening. AB - PURPOSE: To compare the development of tibial tunnel widening after a standard bone-patellar tendon-bone autograft (BPTB) to a flipped BPTB that allows interference screw outlet fixation with a bone plug at both femoral and tibial tunnels, and to identify any observable clinical effect. The hypothesis of this study was that the outlet fixation achieved by the flipped BPTB technique results in diminished tunnel widening at the site of the bone plug. Type TYPE OF STUDY: Nonrandomized control trial. METHODS: The postoperative radiographs of 67 BPTB anterior cruciate ligament (ACL) reconstructions were retrospectively reviewed; 31 had conventional BPTBs and 36 had the bone plugs flipped at the tibial end to achieve interference screw fixation of the bone plug at the tibial outlet as well as the femoral outlet. Biodegradable interference screws (PLLA) were used in all cases, which facilitated tunnel measurements. One week after surgery, the maximal tibial bone tunnel widths were measured on anteroposterior and lateral radiographs 1 cm below the tibial plateau. These initial postoperative measurements were compared with measurement from radiographs taken annually thereafter. Clinical information including Lysholm, Tegner, IKDC activity, Lachman, pivot-shift, and range of motion data was also obtained and compared for the 2 groups. RESULTS: In the conventional BPTB group, at an average follow-up of 28 months, 28 of 31 (90%) showed at least a 2-mm increase (20%) in the tibial tunnel width. The mean maximum tunnel width increase was 2.2 mm. In the flipped BPTB group, at an average follow-up of 31 months, none of these 36 showed any increase in tunnel size. In fact, 15 patients had no distinct tibial tunnel remaining and, of the other 21, the average remaining tunnel width was 3 mm. The clinical results evaluating the Tegner, Lysholm, IKDC activity levels, KT, and physical examination parameters showed no significant differences between these 2 groups. The tunnel width decreased during the first year, but remained unchanged after 12 months. No tunnel dilation was observed with bone plug outlet fixation while 90% of the conventional group had at least a 20% increase (P <.001). CONCLUSIONS: Outlet fixation with the flipped BPTB technique decreased the tibial tunnel width 1 cm below the plateau while 90% of the conventional BPTB patients demonstrated an average widening of 20%. PMID- 12724678 TI - Posterolateral rotatory instability treated by a modified biceps rerouting technique: technical considerations and results in cases with and without posterior cruciate ligament insufficiency. AB - PURPOSE: This study compared the clinical results of modified biceps femoris tenodesis with posterolateral rotatory instability (PLRI) injuries and PLRI combined with PCL injuries. TYPE OF STUDY: Case series. METHODS: Of 46 patients treated for PLRI, 21 had isolated PLRI (group 1), and 25 had PLRI with PCL injuries (group 2). The most common cause of injury was motor vehicle accident. The PCL was reconstructed using an arthroscopic 1-incision technique. The advantages of the modified Clancy technique include fixation of the biceps tendon to the isometric position and reduced surgical damage to the iliotibial band by dissection to the lateral femoral epicondyle through the interval between the iliotibial band and biceps muscle. RESULTS: All knees tested positive in the preoperative reverse pivot shift test, and 43 patients (93%) tested negative postoperatively. The side-to-side difference of an average external rotation thigh-foot angle (ERTFA) at both 30 degrees and 90 degrees of knee flexion was 15 degrees and 11 degrees in group 1 and 21 degrees and 26 degrees in group 2 preoperatively. Postoperative ERTFA tested at 30 degrees and 90 degrees of knee flexion were 10 degrees less than the uninvolved knee in both groups. At a mean follow-up of 40.3 months, the postoperative Lysholm knee score was 93.6 in group 1 and 90.4 in group 2. The postoperative HSS mean value was 91.1 in group 1 and 87.9 in group 2. A correction loss of more than 5 degrees was found in 3 patients of group 1 and in 5 patients of group 2 at an average 12 months after surgery. In 5 of 8 patients, severe scar tissues were found at the insertion site of the biceps tendon to the fibula during surgery. These tissues were associated with damage of the involved structures at the time of injury. CONCLUSIONS: Based on our experience, we recommend the modified biceps tenodesis for the reconstruction of both PLRI injuries and PLRI combined with PCL injuries except in patients with severe damage at the attachment site of the biceps tendon. PMID- 12724679 TI - Patient-controlled bupivacaine infusion into the infrapatellar fat pad after anterior cruciate ligament reconstruction. AB - PURPOSE: The goal of the study was to determine the safety and analgesic efficacy of a patient-controlled infusion pump that dispenses bupivacaine extra articularly to the infrapatellar fat pad after anterior cruciate ligament (ACL) reconstruction with ipsilateral bone-patellar tendon-bone autogenous graft. TYPE OF STUDY: Prospective clinical study. METHODS: Twenty-one consecutive patients were enrolled in the study. After surgery, the catheter of a bupivacaine infusion pump was implanted extra-articularly onto the infrapatellar fat pad. Ten patients were given infusion pumps with 50 mL of 0.5% bupivacaine to be self-administered for analgesia. Eleven patients were given infusion pumps with 50 mL of 0.25% bupivacaine. Opioid analgesics were available if patients were unable to control the pain solely with the bupivacaine pump. Historic controls consisted of 62 consecutive patients who underwent ACL reconstruction before study initiation. Patients recorded volumes of self-administered bupivacaine; also recorded were postoperative pain levels using a visual analog scale (VAS). Opioid analgesic usage was measured and compared with historic controls. RESULTS: Amount of bupivacaine infused and VAS recorded pain levels were not significantly different between the 0.5% and the 0.25% bupivacaine groups. Opioid analgesic usage was significantly reduced when the 0.5% bupivacaine group was compared with historic controls (mean morphine equivalents via intramuscular route: 0.5% group, 17.6 mg; historic controls, 66.4 mg; P =.015). This was also significant when normalized by patient weight (0.5% group, 0.227 mg/kg; historic controls, 0.880 mg/kg; P =.023). Opioid usage in the 0.25% group (35.7 mg and 0.540 mg/kg) was also less than historic controls; however, this was not statistically significant. CONCLUSIONS: Infrapatellar fat pad infusion with 0.5% bupivacaine may provide enhanced analgesia after ACL reconstruction with ipsilateral bone-patellar tendon bone autogenous graft. A double-blinded, randomized control trial with a greater number of patients is required to confirm these findings. PMID- 12724680 TI - Scintigraphic assessment of rabbit knee joints after meniscal allograft transplantation. AB - PURPOSE: It has been shown that meniscal allografts show capsular ingrowth in meniscectomized knees. However, it remains to be established whether a transplanted meniscus can prevent degenerative changes in the long term. In the present study, scintigraphy was used to evaluate degenerative changes in rabbit knees after meniscectomy only and after meniscectomy followed by immediate or delayed meniscus transplantation. TYPE OF STUDY: Experimental study. METHODS: Twenty-eight rabbits were divided into 4 groups. Three rabbits developed infective arthritis and were excluded from the study. In group A (6 animals), 2 rabbits underwent medial meniscectomy, 3 rabbits underwent transplantation with a freshly harvested medial meniscal allograft immediately after meniscectomy, and 1 rabbit underwent a sham operation. In group B (6 rabbits) medial meniscectomy was performed. Group C (6 rabbits) underwent meniscal transplantation immediately after meniscectomy. Group D (7 rabbits) underwent delayed allograft transplantation at 6 weeks after meniscectomy. The animals in group A underwent scintigraphy at 6 weeks after surgery. In the other groups, scintigraphy was performed at 1-year follow-up. Contour changes of the knee joints and uptake of radiolabeled diphosphonate in the subchondral bone were evaluated. RESULTS: No animals in group A showed any abnormalities scintigraphically. Medial compartment changes in group B were more pronounced than in group C, but this difference was not statistically significant. A significant increase in contour changes of the femoral condyle was seen in group D. CONCLUSIONS: Immediate meniscal allograft transplantation did not result in a significant protecting effect on articular cartilage against osteoarthritic degeneration on a long-term basis. Delayed meniscal transplantation revealed even more degenerative changes of articular cartilage than meniscectomy without transplantation. PMID- 12724681 TI - Results of acute arthroscopically repaired triangular fibrocartilage complex injuries associated with intra-articular distal radius fractures. AB - PURPOSE: This study reviews the results of acute repair of peripheral ulnar-sided triangular fibrocartilage complex (TFCC) detachment associated with intra articular distal radius fractures. TYPE OF STUDY: Two-year follow-up of patients who had undergone acute TFCC repair. METHODS: Fifty-six patients underwent arthroscopically assisted treatment of intra-articular distal radius fractures using external fixation and adjunctive percutaneous pinning between 1994 and 1998. Thirteen patients with an acute, complete tear of the ulnar attachment of the TFCC were treated using arthroscopic repair of the TFCC in addition to stabilization of the radius fracture. All patients were evaluated at a mean of 24 months (range, 17 to 35 months) with a physical examination, wrist radiographs, and a Disability of Arm, Shoulder, and Hand (DASH) module outcome assessment questionnaire. RESULTS: Average wrist flexion, extension, pronation, and supination were 67.3, 61.8, 79.1, and 86.8, respectively. The average grip strength was 78% of the uninjured side. The results of the Gartland and Werley grading system were good to excellent in 12 patients and fair in 1 patient. The DASH outcome scores revealed a mean functional score of 13 and a mean athletic score of 12. None of the patients reported ulnar-sided pain at follow-up. CONCLUSIONS: Arthroscopically assisted TFCC repair in conjunction with distal radius fixation resulted in a high degree of patient satisfaction and good to excellent clinical outcomes. PMID- 12724682 TI - Clinical testing for tears of the glenoid labrum. AB - PURPOSE: With the increasing use of shoulder arthroscopy, diagnosis of glenoid labral lesions has become increasingly common. However, a physical examination maneuver that would allow a definitive clinical diagnosis of a glenoid labral tear, and more specifically a SLAP lesion, has been elusive. This study correlated the results of commonly used examination maneuvers with findings at shoulder arthroscopy. The working hypothesis was that 7 commonly used clinical tests, alone or in logical combinations, would provide diagnoses with reliability greater than the accepted standards for magnetic resonance imaging arthrography; i.e., greater than 95% sensitivity and specificity. TYPE OF STUDY: Consecutive sample, sensitivity-specificity study. METHODS: Sixty shoulders undergoing arthroscopy for a variety of pathologies were examined before surgery. All subjects submitted to the Speed test, an anterior apprehension maneuver, Yergason test, O'Brien test, Jobe relocation test, the crank test, and a test for tenderness of the bicipital groove. The examination results were compared with surgical findings and analyzed for sensitivity and specificity in the diagnosis of SLAP lesions and other glenoid labral tears. RESULTS: The results of the O'Brien test (63% sensitive, 73% specific) and Jobe relocation test (44% sensitive, 87% specific) were statistically correlated with presence of a tear in the labrum and the apprehension test approached statistical significance. Performing all 3 tests and accepting a positive result for any of them increased the statistical value, although the sensitivity and specificity were still disappointingly low (72% and 73%, respectively). The other 4 tests were not found to be useful for labral tears, and none of the tests or combinations were statistically valid for specific detection of a SLAP lesion. CONCLUSIONS: Clinical testing is useful in strengthening a diagnosis of a glenoid labral lesion, but the sensitivity and specificity are relatively low. Thus a decision to proceed with surgery should not be based on clinical examination alone. PMID- 12724683 TI - Arthroscopic drilling for chondral, subchondral, and combined chondral subchondral lesions of the talar dome. AB - PURPOSE: We investigated the efficacy of drilling as a treatment for chondral (C), subchondral (S), and combined chondral-subchondral (CS) lesions of the talar dome associated with trauma, using magnetic resonance imaging (MRI), ankle arthroscopy, and the ankle-hindfoot scale of the American Orthopaedic Foot and Ankle Society (AOFAS score). TYPE OF STUDY: Case series study. METHODS: Arthroscopic drilling was performed on 72 patients whose lesions were less than 7 mm in diameter. The patients included 45 men and boys and 27 women and girls whose age at the time of surgery was between 14 and 57 years (mean age, 30.7 +/- 9.5 years). They were followed up for 24 to 71 months (mean follow-up, 39 +/- 6.4 months). RESULTS: There were 13 cases of chondral lesions, 10 cases of S lesions, and 49 cases of CS lesions. The MRI findings revealed that in the chondral lesion group, 13 cases were unchanged and 0 deteriorated; in the S lesion group, 2 improved, 8 were unchanged, and 0 deteriorated; and in the CS lesion group, 13 improved, 36 were unchanged, and 0 deteriorated. The arthroscopic findings showed that in the chondral lesion group, 2 improved, 9 were unchanged, and 1 deteriorated; in the S lesion group, all 8 cases deteriorated; and in the CS lesion group, 28 improved, 22 were unchanged, and 0 deteriorated. Drilling did not always improve the MRI and arthroscopic findings of the 3 respective types of lesions. However, the mean AOFAS score at the most recent follow-up was excellent; 91.7 +/- 2.4 points in the chondral lesion group, 93.1 +/- 2.1 points in the S lesion group, and 98.8 +/- 1.2 points in the CS lesion group. CONCLUSIONS: Our study shows that drilling did not always improve the MRI and arthroscopic findings. However, the clinical results obtained as measured by the AOFAS score were excellent. PMID- 12724684 TI - The disabled throwing shoulder: spectrum of pathology. Part II: evaluation and treatment of SLAP lesions in throwers. PMID- 12724685 TI - Anatomic double-bundle anterior cruciate ligament reconstruction with hamstrings. AB - This article describes a double-bundle gracilis and semitendinosus technique that guarantees a more anatomic anterior cruciate ligament (ACL) reconstruction and allows the surgeon to avoid the use of hardware for graft fixation. The tendons are harvested maintaining their tibial insertion. Sutures are tightened at the free proximal tendon ends to obtain a sufficient strength to traction. The tibial tunnel is located in the medioposterior part of the ACL tibial insertion. For the femoral tunnel, the knee is flexed around 130 degrees and the guide pin is advanced until it passes the femoral cortex. The exit point in the lateral aspect of the femur should be immediately above the end of the lateral femoral condyle. After the lateral incision, the tendons are passed over the top. The correct placement is found by palpating the posterior tubercle of the lateral femoral condyle with a finger. The stitches on the free end of the tendons are tied onto the passing suture that is pulled through the knee joint into the over-the-top position. A suture loop is introduced into the joint through the anteromedial portal using a suture passer and then pulled into the femoral tunnel under the arthroscopic view. The stitches on the free end of the tendons are looped again onto the passing suture, which is pulled through the femoral tunnel, knee joint, and tibial tunnel to retrieve the graft from the tibial incision. The combined gracilis and semitendinosus tendons are then tensioned and secured with a transosseus suture knot. This technique attempts to reproduce the kinematic effect of both anteromedial and posterolateral bundle of the ACL with a 4-bundle reconstruction with a better performance from the anatomic and functional point of view. PMID- 12724686 TI - The radial portal for scaphotrapeziotrapezoid arthroscopy. AB - The scaphotrapeziotrapezoidal (STT) joint is a complex joint in which the arthroscope and instruments can be used through the radial midcarpal and STT ulnar portals. This report describes a new arthroscopic portal at the STT joint to complete and improve the evaluation and treatment of this joint by arthroscopy: The STT-radial (STT-R) portal is situated immediately radial to the abductor pollicis longus at the STT level. Five cadaveric wrist specimens were dissected immediately after the portal was established, and photographed to determine the proximity of neurovascular structures. Magnetic resonance imaging evaluation of 15 hands was performed to measure the proximity of the radial artery. A cadaveric distal scaphoid excision was also created arthroscopically to show the real usefulness of this portal. Results of the anatomic study showed that no lesions of nerves or vessels were seen at this portal. Magnetic resonance imaging showed that the radial artery was located at a safe distance from the portal. The distal scaphoid excision was performed 15 minutes after the STT portals were established. Our results suggest that this new portal is safe and effective. PMID- 12724687 TI - Arthroscopic release of the piriformis muscle under local anesthesia for piriformis syndrome. AB - We developed a minimally invasive technique of releasing the piriformis muscle under endoscopic control for entrapment neuropathy of the sciatic nerve because of tension and contraction of the piriformis muscle. This surgical technique was performed in patients who fulfilled at least 5 of 9 diagnostic criteria we established and who did not respond to conservative therapy for 6 months or more. Although a cavity was maintained using a disposable syringe (10 mL) with a cut tip, an arthroscope (4 mm in diameter) was inserted at an oblique viewing angle of 30 degrees, and the muscle was identified. The area from the musculotendinous junction to the muscle was gradually incised using a special scraper. In particular, pain disappeared simultaneously with release of the piriformis muscle during surgery. With this technique, an adequate cavity can be produced and maintained in a manner similar to that in posterior endoscopic surgery for intervertebral disc herniation. This technique is useful for reducing postoperative pain and allows early return to activity. PMID- 12724689 TI - Cognitive vision, its disorders and differential diagnosis in adults and children: knowing where and what things are. AB - As ophthalmologists we need a basic model of how the higher visual system works and its common disorders. This presentation aims to provide an outline of such a model. Our ability to survey a visual scene, locate and recognise an object of interest, move towards it and pick it up, recruits a number of complex cognitive higher visual pathways, all of which are susceptible to damage. The visual map in the mind needs to be co-located with reality and is primarily plotted by the posterior parietal lobes, which interact with the frontal lobes to choose the object of interest. Neck and extraocular muscle proprioceptors are probably responsible for maintaining this co-location when the head and eyes move with respect to the body, and synchronous input from both eyes is needed for correct localisation of moving targets. Recognition of what is being looked at is brought about by comparing the visual input with the "image libraries" in the temporal lobes. Once an object is recognised, its choice is mediated by parietal and frontal lobe tissue. The parietal lobes determine the visual coordinates and plan the visually guided movement of the limbs to pick it up, and the frontal lobes participate in making the choice. The connection between the occipital lobes and the parietal lobes is known as the dorsal stream, and the connection between the occipital lobes and the temporal lobes, comprises the ventral stream. Both disorders of neck and extraocular muscle proprioception, and disorders leading to asynchronous input along the two optic nerves are "peripheral" causes of impaired visually guided movement, while bilateral damage to the parietal lobes can result in central impairment of visually guided movement, or optic ataxia. Damage to the temporal lobes can result in impaired recognition, problems with route finding and poor visual memory. Spontaneous activity in the temporal lobes can result in formed visual hallucinations, in patients with impaired central visual function, particularly the elderly. Deficits in cognitive visual function can occur in different combinations in both children and adults depending on the nature and distribution of the underlying brain damage. In young children the potential for recovery can lead to significant improvement in parietal lobe function with time. Patients with these disorders need an understanding of their deficits and a structured positive approach to their rehabilitation. PMID- 12724688 TI - The PC knot: a secure and satisfying arthroscopic slip knot. AB - In an attempt to attain equivalent results in arthroscopic versus open shoulder and knee soft tissue reconstructions, orthopaedic surgeons require reproducible techniques and quality equipment. A new arthroscopic slip knot is proposed: the PC knot. The optimal arthroscopic slip knot must be (1) low profile, (2) easy to throw, (3) slide well, (4) easy to set, and (5) provide excellent initial security for reliable soft tissue apposition. The PC knot satisfies these criteria and offers variable loop tensioning that allows for improved slidability and initial loop security. PMID- 12724690 TI - Molecular genetics of microvascular disease in diabetic retinopathy. AB - Diabetic retinopathy is a sight-threatening complication of the retinal microvasculature. While important environmental factors have been clearly identified as influencing its development, increasing evidence suggests that diabetic retinopathy has a genetic component. A variety of studies have explored associations between candidate genes and frequency and severity of retinopathy. Overall, this review has found that the majority of candidate genes studied exhibit weak or no association with retinopathy status, and where associations have been detected these results have not been replicated in multiple populations. This may reflect inaccurate case definition, small subject numbers and possibly inadequate markers for genetic studies. PMID- 12724691 TI - Leber's hereditary optic neuropathy triggered by antiretroviral therapy for human immunodeficiency virus. AB - PURPOSE: To describe the clinical features of two cases of Leber's hereditary optic neuropathy (LHON) precipitated by antiretroviral treatment for human immunodeficiency virus (HIV) infection. METHODS: Two cases of LHON (from an expected four new cases a year throughout Australia) were identified in men on treatment for HIV infection. RESULTS: Two HIV-infected men were receiving combination antiretroviral therapy that included nucleoside analogues. Both patients carried the 14 484 mitochondrial DNA mutation and were distantly related (seventh cousins). Although both men presented with sequential visual loss typical of LHON and one had a known close relative affected by LHON, the correct diagnosis was delayed in both cases. The final visual outcome was profoundly reduced in both instances and cessation of antiretroviral therapy did not result in recovery of vision in one patient. CONCLUSION: Patients with a family history of LHON who require antiretroviral treatment should be warned of the high risk of severe visual loss. The underlying mechanism of antiretroviral side effects may help characterize the other trigger factors for LHON. PMID- 12724692 TI - Visual complications of proton beam therapy for clival chordoma. AB - PURPOSE: Combining maximal surgical resection with high-dose proton radiation therapy is reported to be currently the best management of patients with clival chordoma. METHOD: Since 1991, four consecutive patients from our institution with this tumour have been referred for postoperative proton beam radiotherapy. RESULT: We have experienced an unusually high complication rate (50%) of delayed radiation optic neuropathy. This has resulted in profound, bilateral visual loss at 1 and 2 years postproton beam treatment. CONCLUSION: It has served as a reminder that proton beam therapy is not an innocuous treatment option and this devastating complication should be taken into account when discussing the management of clivus chordoma. We postulate whether the optic apparatus is particularly sensitive to proton vs photon damage. PMID- 12724694 TI - Does frequency doubling technology perimetry reliably detect neurological visual field defects? AB - AIM: To determine the ability of frequency doubling technology (FDT) perimetry to detect visual field defects of neurological origin. METHODS: A total of 15 eyes of nine patients who all had complete hemianopias or quadrantanopias underwent the FDT 20-5 screening mode test and Humphrey 24-2 SITA Fast visual field test (HFA). The FDT results were scored according to the number of abnormal test locations (out of a maximum of 4) in each affected quadrant. FDT locations showing a defect of P< 2% were considered abnormal. RESULTS: Of the 15 eyes, six showed complete superior quadrantanopic and nine complete hemianopic field defects on HFA. Of 96 FDT test locations in these quadrants or hemifields only 38 were abnormal on FDT testing (40%). For the quadrantanopic field defects, five out of 24 locations were abnormal (21%). For the hemianopic field defects, 33 out of 72 locations were abnormal (49%). In three eyes (two with quadrantanopias and one with complete hemianopia), FDT perimetry failed to demonstrate any corresponding abnormality. CONCLUSIONS: The FDT screening test can fail to demonstrate complete hemianopic and quadrantanopic field defects. Users should be aware of this deficiency when using FDT to screen for field defects. PMID- 12724693 TI - Quantitative evaluation of the retinal venous tortuosity in chronic anaemic patients affected by beta-thalassaemia major. AB - AIMS: Retinal venous tortuosity (RVT) is a common finding in patients affected by different forms of chronic anaemia. The aims of this study were to quantify RVT in anaemic patients with beta-thalassaemia major and to verify whether it is related to some of the following parameters: patient's age, ferritin plasma level, and Desferrioxamine (DFX) daily dosage. METHODS: A retrospective study was carried out. In total, 36 consecutive thalassaemic patients, treated with polytransfusion regimen and DFX, were age- and sex-matched with a control group of 36 normal subjects. All subjects bilaterally underwent red-free fundus photography, centred on the optic disc. The four main retinal veins were measured with a computer-assisted method. RESULTS: Mean venous length in the thalassaemic group was significantly greater than that observed in the control group (P <0.001). In thalassaemic patients, no significant correlations between retinal venous length and, respectively, plasma ferritin level and DFX daily dosage were documented. Statistical analysis demonstrated a very significant association between patient's age and increased RVT only in thalassaemic patients (P <0.001). CONCLUSIONS: Our findings demonstrate that patients with beta-thalassaemia major have increased RVT, as compared to normal subjects. In this selected anaemic population, patient's age, closely related to anaemia duration, is the only variable responsible for the RVT increment. This clinical sign indicates a long standing duration of anaemia. PMID- 12724695 TI - Surgical correction of large-angle exotropia in adults. AB - AIM: A retrospective and longitudinal review of the outcome of strabismus surgery for adults with large- and very-large-angle manifest exodeviations, using two-, three- and four muscle horizontal recti surgery with adjustable sutures. METHODS: A total of 26 consecutive adult patients undergoing surgery for socially noticeable strabismus comprising five primary, 16 consecutive, and five secondary constant exotropias with a mean near deviation of 58 prism dioptres and a mean distance deviation of 55Delta were evaluated preoperatively and at various time intervals postoperatively. Surgery involved two muscles in seven cases, three muscles in 13 cases, and four muscles in six cases; and 25 of 26 had adjustable sutures. There was a horizontal preoperative ocular movement deficit in 17 that was asymmetrical in four cases. RESULTS: Binocularity was restored in eight patients (31%), 20 (77%) were within 10Delta of orthotropia, and 24 (92%) were happy with their cosmesis. Two had symptomatic asymmetrical ocular motility deficits postoperatively following a two-muscle procedure and one required reoperation. A total of 19 patients undergoing three- or four-muscle surgery were asymptomatic postoperatively. A total of 22 patients had follow-up of 8 months or more. CONCLUSION: In adults with large-angle manifest exodeviations, adjustable suture surgery involving three or more horizontal recti successfully restores primary position alignment, a high degree of patient satisfaction, and can be expected to be associated with a low incidence of symptomatic postoperative asymmetrical ocular movement deficits. PMID- 12724696 TI - UBM-guided chamber angle surgery for glaucoma management: an experimental study. AB - PURPOSE: The aim of this experimental study was to investigate the potential of ultrasound bimicroscopy (UBM)-guided chamber angle surgery as an alternative or supplement to gonioscopy and intraocular microendoscopy for intraoperative control. METHODS: In 15 porcine cadaver eyes, mechanical goniopuncture or punctual Er:YAG laser trabecular ablation was performed without operating microscope or gonioscopy, but with real-life ultrasound biomicroscopy monitoring with a 50 MHz transducer. Intraoperative localization of the microsurgical instruments and tissue-instrument contact were qualitatively evaluated. RESULTS: The instruments could be clearly visualized within the chamber angle and disturbing artefacts were only minimal when using mechanically fixed instruments in slow motion. Topographic localization, tissue contact, and penetration depth of the instruments entering the scleral were well illustrated as far as the technical resolution limits of UBM would allow. CONCLUSIONS: UBM can be used intraoperatively to monitor the correct manoeuvring of microsurgical instruments in selected ab interno procedures. Some adaptations and further modifications of the technique presented here will be necessary before UBM-guided surgery can be considered for clinical use in humans. PMID- 12724697 TI - Results of cataract surgery in renal transplantation patients. AB - PURPOSE: To evaluate the results of conventional extracapsular cataract extraction and phacoemulsification cataract surgery, in renal transplant patients. METHODS: A total of 11 eyes of eight renal transplant recipients who underwent cataract surgery in Ankara between 1995 and January 2001 were evaluated. Each individual underwent routine ophthalmologic examination including visual acuity with Snellen chart testing, slit-lamp examination, intraocular pressure measurement, and fundus examination. Of these, seven patients had posterior subcapsular cataracts and four had nuclear cataracts. Conventional extracapsular cataract extraction (cECCE) was performed in four eyes with single piece all-polymethylmethacrylate posterior chamber intraocular lens implantation. The other seven eyes were treated with small-incision phacoemulsification and implantation of a foldable hydrophilic acrylic lens. RESULTS: We observed intraoperative posterior capsule rupture and vitreous loss in one patient, postoperative intraocular tension elevation in one patient, and sterile suture infiltration in one patient in our series. Average of the visual acuities 6 months after the operation was 20/30 (range, 20/60-20/20). CONCLUSIONS: In this, the first published report to describe outcomes of cataract surgery in renal transplant patients, most complications were associated with high of steroids use and immunosuppressives, and were independent of the type of cataract surgery used, either ECCE or phacoemulsification surgery. The study suggests the need for additional care before, during, and after operation. PMID- 12724698 TI - Serum leptin concentrations are decreased and correlated with disease severity in age-related macular degeneration: a preliminary study. AB - BACKGROUND: Age-related maculopathy (ARM) or degeneration (ARMD) is the leading cause of irreversible blindness in developed countries. Despite several studies on the morphology of ARMD, the aetiology is unknown and factor(s) contributing to the pathogenesis remain to be characterised. More recent studies have demonstrated that cholesterol esters and lipids are present within Bruch's membrane deposits and drusen, and dietary fat intake is associated with ARMD. The product of Ob gene, leptin, is a recently discovered peptide participating in human metabolism. There is a direct relationship between serum leptin and diet, and lipoprotein metabolism, but the role of leptin in the course of ARMD has not previously been investigated. PURPOSE: This cross-sectional case-control study investigated whether serum leptin level was associated with ARMD as a new possible risk factor and to assess its relationship with disease severity. Methods A total of 32 patients with ARM or ARMD (17 men, 15 women) and 20 age- and sex-matched healthy control subjects without ARMD (11 men, nine women) from a similar ethnic background were enrolled in this multicentre study. Body mass index (BMI) (weight (kg)/height (m(2))) was calculated for each group. The presence of maculopathy was assessed on the basis of colour fundus photographs using an international classification system. Patients were classified as early ARM (n=16) or late-ARMD (n=16) using clinical examination and grading of photographs. Serum leptin levels were measured by an enzyme-linked immunosorbent assay kit. The Mann-Whitney U test or chi(2) test was used for statistics as indicated, and P&<0.05 was considered to be significant. RESULTS: The age, sex ratio, and BMI between groups were comparable. Patients with maculopathy had significantly (P&<0.001) lower leptin levels (mean+/-SD, 6.01+/-2.55 ng/ml) than control subjects (13.21+/-2.27 ng/ml). In addition, late-ARMD patients had significantly lower leptin levels (3.81+/-0.58 ng/ml) than early-ARM patients (8.21+/-1.68 ng/ml, P&<0.001) or control subjects (P&<0.001). CONCLUSION: Leptin seems to be a possible newly associated factor in the course of ARM and may be involved in the lipid composition of the macular lesions, especially in late ARMD. PMID- 12724699 TI - Epidemic kerato-conjunctivitis--do outbreaks have to be epidemic? AB - PURPOSE: To study: the epidemiology of an outbreak of adenoviral keratoconjunctivitis in a UK teaching hospital; disease presentation and its effect on clinical diagnostic efficiency; patterns of viral transmission between staff and patients; the effectiveness of infection control procedures in minimising outbreaks. METHODS: Prospective/retrospective clinical audit and retrospective audit of virological culture results: all viral culture swabs taken during an outbreak of adenoviral keratoconjunctivitis were analysed. The case records of patients whose viral swabs were positive for adenoviral culture were traced. The time for viral cultures to become positive (culture positive time) was calculated. Analysis of the case notes was performed to elucidate (1) the source of infection and (2) the risk factors for acquisition of the infection. Retrospective clinical audit was performed to evaluate the effectiveness of infection control procedures. Adenovirus isolates underwent serotyping. RESULTS: During the 3-month period of study, there were 38 confirmed cases of adenoviral keratoconjunctivitis. This represented a 217% increase in the number of new cases per 3-month period. The case notes for five patients were untraceable. Of the remaining 33 patients, 21 (63%) had acquired their infection either directly or indirectly from the eye department and 22 (67%) had presented with unilateral disease. The rate of misdiagnosis was higher (9/22=42%) in patients presenting with unilateral disease than those presenting with bilateral disease (2/11=18%). Intradepartmental acquisition of infection was associated with invasive procedures, for example use of diagnostic/therapeutic contact lenses. Culture positive times ranged from 3 to 29 days. The introduction of infection control procedures was associated with a dramatic decrease in the incidence of departmentally acquired cases with no new cases after 2 weeks. Multiple serotypes of adenovirus were involved. CONCLUSION: Outbreaks of adenoviral keratoconjunctivitis are a serious public health issue concerning ophthalmic departments. This audit study illustrates several important points: (1) how hospital-acquired infection can account for a significant proportion of the cases seen, (2) how multiple types of adenovirus can be involved in a single outbreak, (3) that severe unilateral disease is associated with a higher rate of misdiagnosis; and (4) how standard viral culture techniques may not be satisfactory in confirming/disproving infection when the diagnosis is in doubt. The potential benefit of infection control procedures in minimising this outbreak could not be proven within this audit. PMID- 12724700 TI - Vasoproliferative tumours of the retina. AB - This paper describes three cases of vasoproliferative tumours of the retina including histopathology in one. The clinical presentation, differential diagnosis and treatment modalities are discussed with a brief review of the literature. PMID- 12724701 TI - Visual function in the brain-damaged child. AB - The essential role of the primary visual cortex in visual processing has been extensively studied over the last century or more. Injuries to the visual cortex in adult humans can produce blindness, referred to as "cortical blindness". In children some degree of visual recovery has been noted in comparable injuries and for that reason the term "cortical visual impairment" has been suggested as a more appropriate diagnosis in children. This term is, however, inaccurate as a significant number of children with visual loss and neurologic damage have injuries to the noncerebral pathways (for example--optic radiations in children with periventricular leukomalacia). In this study we compare visual outcomes and recovery in children with primary visual cortex lesions vs those with periventricular leukomalacia. We suggest that the poorer outcomes of children with periventricular leukomalacia could have been predicted based on studies of the mechanisms of visual recovery in infant animals following visual cortex ablation. PMID- 12724702 TI - Corneal replacement using a synthetic hydrogel cornea, AlphaCor: device, preliminary outcomes and complications. AB - PURPOSE: Clinical assessment of outcome of corneal replacement with a synthetic cornea, AlphaCor, in patients considered at too high risk for conventional penetrating keratoplasty with donor tissue to be successful, but excluding indications such as end-stage dry eye that might be suited to traditional prosthokeratoplasty. METHODS: All patients in the multicentre clinical trial were managed according to an approved protocol, with Ethics Committee approval in each centre. Preoperative visual acuity ranged from perception of light (PL) to 6/60 (20/200). Implantation was by means of an intralamellar technique, with a conjunctival flap in most cases. Tissues anterior to the optic were removed as a secondary procedure. RESULTS: Up to 30 November 2001, 40 AlphaCor devices had been implanted in 38 patients, of mean age 60 years. Follow-up ranged from 0.5 months to 3 years. There had been one extrusion (2.5%) and four cases (10%) where a device had been removed due to melt-related complications. All five of these cases received a donor corneal graft after the device was removed, with these grafts remaining anatomically satisfactory and epithelialised to date. Corneal melts in AlphaCor recipients were found to be strongly associated with a history of ocular herpes simplex infection. Two further devices (5%) were removed owing to reduced optic clarity after presumed drug-related deposition, and have been successfully replaced with second devices. Mean preoperative best-corrected visual acuity was hand movements. Visual acuities after surgery ranged from PL to 6/6(-2) (20/20(-2)). CONCLUSIONS: Early results suggest that the AlphaCor, previously known as the Chirila keratoprosthesis (Chirila KPro), has a low incidence of the complications traditionally associated with keratoprostheses and can be effective in restoring vision in patients considered untreatable by conventional corneal transplantation. Importantly, the device can be replaced with a donor graft in the event of development of a significant complication. A history of ocular herpes simplex is a contraindication to AlphaCor implantation. Ongoing monitoring of clinical outcomes in all patients will allow the indications for AlphaCor, as opposed to donor grafts, to be determined. PMID- 12724703 TI - Calcification of modern foldable hydrogel intraocular lens designs. AB - PURPOSE: To report and compare clinical and pathological features of hydrophilic acrylic intraocular lenses (IOLs) of three major designs, explanted from patients who had visual disturbances caused by opacification of the lens optic. METHODS: Eighty-seven hydrophilic acrylic IOLs (25 Hydroview, 54 SC60B-OUV, and 8 Aqua Sense lenses) were explanted and sent to our center. Most patients became symptomatic during the second year after cataract surgery. A fine granularity was observed on the surface of the lens optic in the case of Hydroview. With the SC60B-OUV and Aqua-Sense lenses, the opacity resembled a nuclear cataract. Gross examination, light microscopy and staining with alizarin red and the von Kossa method (for calcium) were performed. Some lenses were submitted for scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS). RESULTS: Light microscopic and SEM analyses revealed the presence of irregular granular deposits on the external optical surfaces of Hydroview lenses. With the SC60B-OUV lenses, the opacity was caused by the presence of multiple fine, granular deposits within the lens optic, distributed in a line parallel to the anterior and posterior curvatures of the optic, with a clear zone just beneath its external surfaces. The Aqua-Sense lenses exhibited both patterns simultaneously. The deposits in all cases stained positive with alizarin red and von Kossa method. EDS also demonstrated the presence of calcium and phosphates within the deposits. CONCLUSION: Differences in the water content of the hydrophilic acrylic materials used in the manufacture of these three lens designs may be responsible for the different patterns of calcium precipitation. Careful clinical follow up of patients implanted with these lenses is necessary to determine if this phenomenon is rare and sporadic or may be more widespread. PMID- 12724704 TI - Subjective visual experience during phacoemulsification cataract surgery under sub-Tenon's block. AB - In a prospective survey, 121 patients received sub-Tenon's block and were interviewed on the same day after their phacoemulsification cataract surgery regarding their visual experience in the operated eye during surgery. Majority of patients (81%) reported that they could see some light during surgery and various colours were seen by 56%. Movements of various descriptions were also reported by 40% of patients. The majority of patients (93%) found the visual experience acceptable but 4% thought it to be unpleasant and 3% found it frightening. Patients undergoing phacoemulsification surgery under sub-Tenon's block experience a variety of visual sensations and some patients may be frightened. All patients should receive appropriate preoperative warning. PMID- 12724705 TI - Free-floating melanin particles in the anterior chamber: a normal finding in African eyes? AB - PURPOSE: Pigment dispersion syndrome (PDS) is a well-described entity with Krukenberg's spindle, heavy trabecular pigmentation and retroilluminating iris defects. We have observed a group of patients in mesoendemic onchocercal communities of Kaduna State, Nigeria, with significant amounts of free-floating melanin in the anterior chamber, normal angle pigmentation and absence of iris defects. A pseudo-Krukenberg spindle forms when the patients are asked to maintain a 2 min head-down posture as is often done when examining eyes for the presence of anterior chamber microfilaria. This spindle gradually disappears (tumbles back) after about 2 min of return to the erect posture. This paper describes this finding, which does not appear to fit into accepted notions of pigment dispersion. METHODS: As part of the seventh annual ivermectin dosing exercise during which evidence of optic nerve damage was sought, 455 patients were examined for the presence of microfilaria in the anterior chamber. A total of 352 had been selected for signs of optic nerve disease during an earlier screening exercise, while 103 belonged to a random sample of 5 years and above. Signs of onchocerciasis were sought, while gonioscopy and intraocular pressure measurements were carried out. RESULTS: Of the 455 (11%) individuals examined, 53 demonstrated this phenomenon. Within the random sample, the prevalence was 20%. These tumbling Krukenberg positive (TK+) individuals are significantly younger than TK- individuals and the prevalence, highest in the first decade, dropped steadily to zero levels over the age of 75. Sex distribution was about equal. There was no difference in intraocular pressure, cup-disc ratio and angle pigmentation. Distributions of sclerosing keratitis and chorioretinitis were not statistically different. Optic nerve disease was more common in TK- but this was attributable to the older age distribution. Five TK+ were re-examined after a period of 7 years and had not developed PDS or glaucoma. Four of the five remained TK+. A familial tendency was noted and hereditary factors may be at play, possibly autosomal recessive. The same phenomenon was noted in two of 44 patients in an ophthalmic clinic in Abuja, Nigeria, an urban, non-endemic city south of Kaduna. CONCLUSIONS: This phenomenon does not fit into accepted notions of PDS and may well be a normal finding. PMID- 12724706 TI - The use of ultrasound as an aid in the diagnosis of giant cell arteritis: a pilot study comparing histological features with ultrasound findings. AB - AIMS: We present our preliminary experience with the use of ultrasound in aiding the diagnosis of giant cell arteritis (GCA). Schmidt et al have previously described a hypoechoic or "halo" effect surrounding the walls of affected arteries on examination with ultrasound. We illustrate these features and explore the attributes and limitations of this technique. METHOD: Two groups of patients were recruited: (1) patients with suspected GCA awaiting temporal artery biopsy and (2) patients with no history or symptoms of GCA of a similar age group. All the recruited patients underwent ultrasound examination of both temporal arteries. The findings on ultrasound were compared with the results of the histological specimens in group 1. For this study, the histological findings alone were used to define if a patient was suffering from GCA. No biopsies were taken in the patients in group 2. RESULTS: Out of 26 patients with suspected GCA, seven patients were found to be positive on biopsy, of which six had been identified on ultrasound. Six patients were found to be false positive on ultrasound, but all had moderate-to-severe features of arteriosclerosis on histology. A total of 13 patients were found to be negative on ultrasound and negative on biopsy for GCA, two of these patients had histological features of arteriosclerosis. In the group with no symptoms of GCA (12 patients), in two patients hypoechoic areas were detected. The results presented give a sensitivity of 86%, specificity of 68%, and positive predictive value of 50% for the use of ultrasound in the diagnosis of GCA. CONCLUSIONS: This preliminary study indicates that this test may be helpful in those patients with symptoms suggestive of GCA, but currently we cannot recommend any change of present practice. PMID- 12724707 TI - A case of cutaneous melanoma metastatic to the right eye and left orbit. PMID- 12724709 TI - Primary squamous cell carcinoma of the lacrimal gland. PMID- 12724708 TI - Fluorescein angiography and patchy skin discoloration: a case report. PMID- 12724710 TI - Choroidal melanoma presenting as a haemorrhagic detachment in a 12-year old. PMID- 12724711 TI - Uric acid crystals presenting as an orbital mass. PMID- 12724712 TI - A rare solitary fibrous tumour of the lacrimal sac presenting as acquired nasolacrimal duct obstruction. PMID- 12724713 TI - Reduced visual acuity following standard ETDRS macular laser for clinically significant macular oedema. PMID- 12724714 TI - Ophthalmia nodosa secondary to tarantula hairs. PMID- 12724715 TI - A case of orbital myositis secondary to orbital cellulitis in a child. PMID- 12724716 TI - Associations of high hypermetropia in childhood. PMID- 12724717 TI - Choriovitreal neovascularization following transpupillary thermotherapy for choroidal melanoma. PMID- 12724718 TI - Massive spontaneous expulsive suprachoroidal haemorrhage in a blind glaucomatous eye treated with chronic topical steroid. PMID- 12724719 TI - Microbial keratitis following orthokeratology. PMID- 12724720 TI - Aqueous misdirection following needling of trabeculectomy bleb. PMID- 12724721 TI - Systemic side effects of topical latanoprost. PMID- 12724722 TI - Cutaneous latanoprost in the treatment of alopecia areata. PMID- 12724723 TI - Corneal adherent leukoma associated with measles. PMID- 12724724 TI - Intraocular cysticercosis simulating retinoblastoma in a 5-year-old child. PMID- 12724725 TI - Scleromalacia as a complication of herpes zoster ophthalmicus. PMID- 12724726 TI - Pseudophakic posterior iris chafing syndrome. PMID- 12724727 TI - Debris on processed ophthalmic instruments: a cause for concern. PMID- 12724729 TI - Giant mucocoele masquerading as chronic unilateral conjunctivitis. PMID- 12724730 TI - Translational strategies for cancer prevention in liver. AB - Unlike many other types of human cancer, the aetiology of liver cancer is well understood. Infection with hepatitis viruses, coupled with dietary exposure to the fungal toxin aflatoxin, increases the risk of the disease. Although primary prevention, based on vaccination and avoiding exposure to these agents, is an appealing option, such strategies will require considerable investment of time and resources to be successful. In the developing world--where the burden of liver cancer is highest--immediate, practical and economical approaches are essential. So, targeted chemoprevention might be most appropriate for the present generation of individuals at risk. PMID- 12724731 TI - 5-fluorouracil: mechanisms of action and clinical strategies. AB - 5-fluorouracil (5-FU) is widely used in the treatment of cancer. Over the past 20 years, increased understanding of the mechanism of action of 5-FU has led to the development of strategies that increase its anticancer activity. Despite these advances, drug resistance remains a significant limitation to the clinical use of 5-FU. Emerging technologies, such as DNA microarray profiling, have the potential to identify novel genes that are involved in mediating resistance to 5-FU. Such target genes might prove to be therapeutically valuable as new targets for chemotherapy, or as predictive biomarkers of response to 5-FU-based chemotherapy. PMID- 12724732 TI - Cancer and ageing: rival demons? AB - Organisms with renewable tissues use a network of genetic pathways and cellular responses to prevent cancer. The main mammalian tumour-suppressor pathways evolved from ancient mechanisms that, in simple post-mitotic organisms, act predominantly to regulate embryogenesis or to protect the germline. The shift from developmental and/or germline maintenance in simple organisms to somatic maintenance in complex organisms might have evolved at a cost. Recent evidence indicates that some mammalian tumour-suppressor mechanisms contribute to ageing. How might this have happened, and what are its implications for our ability to control cancer and ageing? PMID- 12724734 TI - Tumour-cell invasion and migration: diversity and escape mechanisms. AB - Cancer cells possess a broad spectrum of migration and invasion mechanisms. These include both individual and collective cell-migration strategies. Cancer therapeutics that are designed to target adhesion receptors or proteases have not proven to be effective in slowing tumour progression in clinical trials--this might be due to the fact that cancer cells can modify their migration mechanisms in response to different conditions. Learning more about the cellular and molecular basis of these different migration/invasion programmes will help us to understand how cancer cells disseminate and lead to new treatment strategies. PMID- 12724733 TI - The circadian clock: pacemaker and tumour suppressor. AB - The circadian rhythms are daily oscillations in various biological processes that are regulated by an endogenous clock. Disruption of these rhythms has been associated with cancer in humans. One of the cellular processes that is regulated by circadian rhythm is cell proliferation, which often shows asynchrony between normal and malignant tissues. This asynchrony highlights the importance of the circadian clock in tumour suppression in vivo and is one of the theoretical foundations for cancer chronotherapy. Investigation of the mechanisms by which the circadian clock controls cell proliferation and other cellular functions might lead to new therapeutic targets. PMID- 12724735 TI - Cancer revoked: oncogenes as therapeutic targets. AB - Recent findings show that even the brief inactivation of a single oncogene might be sufficient to result in the sustained loss of a neoplastic phenotype. It is therefore possible that the targeted inactivation of oncogenes could be a specific and effective treatment for cancer. So why does oncogene inactivation cause tumour regression and will this be a generally successful approach for the treatment of human neoplasia? PMID- 12724736 TI - Photodynamic therapy for cancer. AB - The therapeutic properties of light have been known for thousands of years, but it was only in the last century that photodynamic therapy (PDT) was developed. At present, PDT is being tested in the clinic for use in oncology--to treat cancers of the head and neck, brain, lung, pancreas, intraperitoneal cavity, breast, prostate and skin. How does PDT work, and how can it be used to treat cancer and other diseases? PMID- 12724737 TI - Solid response to SARS--almost. PMID- 12724738 TI - NO adducts in mammalian red blood cells: too much or too little? PMID- 12724740 TI - C75 inhibits food intake by increasing CNS glucose metabolism. PMID- 12724741 TI - SARS epidemic unmasks age-old quarantine conundrum. PMID- 12724742 TI - Mosquito mating game could mean buzzkill for Brits. PMID- 12724744 TI - Hollywood horrors bring bioterror to life. PMID- 12724745 TI - New wave of AIDS rocks Pacific Islands. PMID- 12724746 TI - Ozone exposure throws monkey wrench into infant lungs. PMID- 12724748 TI - Goats to de-liver cells for transplants in India. PMID- 12724749 TI - UK launches tumor bank to match maligned Biobank. PMID- 12724750 TI - Roy Anderson. PMID- 12724751 TI - Building the translational highway: toward new partnerships between academia and the private sector. PMID- 12724752 TI - Nitric oxide's reactions with hemoglobin: a view through the SNO-storm. PMID- 12724755 TI - Organ alchemy: producing insulin in the liver. PMID- 12724756 TI - Mysticism to medicine. PMID- 12724757 TI - Calcium and heart failure: the cycle game. PMID- 12724758 TI - Mining the genome for combination therapies. PMID- 12724759 TI - Platelet attraction. PMID- 12724760 TI - Mastering innate immunity. PMID- 12724761 TI - Cisplatin and the sensitive cell. PMID- 12724763 TI - Novel strategies for the treatment of sepsis. AB - The history of therapeutic interventions in clinical trials for sepsis has been referred to as the "graveyard for pharmaceutical companies." That is now set to change, as research provides hope for new approaches that will be therapeutically effective in humans with sepsis. PMID- 12724765 TI - Science for the people. PMID- 12724766 TI - The N-end rule and regulation of apoptosis. PMID- 12724767 TI - Morphogens recycled. PMID- 12724768 TI - Split personalities: the agonistic antagonist Sprouty. PMID- 12724769 TI - Monopolar attachment by Polo. PMID- 12724770 TI - Sliding doors: clathrin-coated pits or caveolae? PMID- 12724771 TI - Deafening cycle. PMID- 12724772 TI - Small changes in Wnt signalling. PMID- 12724773 TI - Chromosomes, positions please! PMID- 12724774 TI - Memorable transcription. PMID- 12724776 TI - Chromatin and transcription: histones continue to make their marks. PMID- 12724781 TI - NTP Toxicology and Carcinogenesis Studies of Tetracycline Hydrochloride (CAS No. 64-75-5) in F344/N Rats and B6C3F1 Mice (Feed Studies). AB - Tetracycline hydrochloride is a broad-spectrum antibiotic used for its bactericidal action in human and veterinary medicine. Toxicology and carcinogenesis studies of tetracycline hydrochloride (USP grade, 91% pure) were conducted by feeding diets containing tetracycline hydrochloride to groups of F344/N rats and B6C3F1 mice of each sex for 14 days, 13 weeks, or 2 years. Fourteen-Day and Thirteen-Week Studies: The same dietary concentrations were used for the 14-day and 13-week studies (0, 3,125, 6,250, 12,500, 25,000 and 50,000 ppm tetracycline hydrochloride). In the 14-day studies, none of the rats or mice died. The final mean body weight of male rats that received 50,000 ppm was 24% lower than that of the controls. The final mean body weight of mice that received 50,000 ppm in the diet was 18% lower than that of the controls for males and 15% lower for females. No compound-related effects were observed in rats or mice at necropsy. During the 13-week studies, none of the rats or mice died. The final mean body weight of male rats that received 50,000 ppm was 18% lower than that of the controls. Compound-related effects included cytoplasmic vacuolization in the liver of male rats at 25,000 and 50,000 ppm. Bone tetracycline concentrations in rats and mice increased with increasing dose of tetracycline hydrochloride. The final mean body weight of mice that received 50,000 ppm was 16% lower than that of the controls for males and 6% lower for females. Estimated feed consumption by dosed rat and mouse groups was similar to that of the controls. No compound related gross or microscopic pathologic effects were observed in mice. Based on these results, 2-year studies of tetracycline hydrochloride were conducted by feeding diets containing 0, 12,500, or 25,000 ppm tetracycline hydrochloride to groups of 50 rats and 50 mice of each sex for 103 weeks. Body Weight, Survival, and Feed Consumption in the Two-Year Studies: Mean body weights of dosed and control male and female rats were similar throughout most of the studies. The survival of both the low and high dose female groups was greater than that of the controls. No significant differences in survival were observed between any groups of male rats (male: control, 27/50; low dose, 24/50; high dose, 31/50; female: 27/50; 39/50; 38/50). Mean body weights of dosed mice were markedly (more than 10%) lower than those of the controls throughout most of the studies. The survival rates of the dosed groups of male mice were greater than that of the control group. No significant differences in survival were observed between any groups of female mice (male: 31/50; 43/50; 43/50; female: 37/50; 35/50; 38/50). Feed consumption was similar by dosed and control rats and mice of either sex throughout the studies. Nonneoplastic and Neoplastic Effects in the Two-Year Studies: Basophilic cytoplasmic change and clear cell change were positively correlated with tetracycline hydrochloride administration in male rats. Otherwise, no significant increases in neoplastic or nonneoplastic lesions in rats or mice of either sex were considered related to tetracycline hydrochloride administration. The incidence of adenomas or carcinomas (combined) of the pancreatic islets in low dose male rats was greater than that in the controls (control, 0/49; low dose, 5/49; high dose, 0/49). This marginal effect in the low dose group was not considered to be chemically related. The historical control rate of pancreatic islet cell neoplasms from previous studies at this laboratory is 6% (9/148). Decreased incidences and severity of chronic nephropathy in male rats were associated with tetracycline hydrochloride administration (48/50; 35/50; 36/50). Female mice administered tetracycline hydrochloride in feed did not develop hepatocellular adenomas or carcinomas (combined incidence: 10/49; 0/48; 0/50). The historical control rate for hepatocellular adenomas or carcinomas (combined) from previous studies at this laboratory is 18/149 (12%). Other decreases in tumor incidence involving several tissues were considered to be of ma tumor incidence involving several tissues were considered to be of marginal biologic significance. Genetic Toxicology: Tetracycline hydrochloride was not mutagenic in four strains of Salmonella typhimurium (TA98, TA100, TA1535, or TA1537) when tested in a preincubation protocol in the presence or absence of exogenous metabolic activation. Tetracycline hydrochloride was negative in the mouse lymphoma L5178Y/TK± assay with or without induced rat liver S9 but gave a marginally positive response when tested in the presence of noninduced S9. In cytogenetic assays with Chinese hamster ovary (CHO) cells, treatment with tetracycline hydrochloride, both with and without S9, did not induce chromosomal aberrations or sister chromatid exchanges (SCEs). Tetracycline hydrochloride did not induce sex-linked recessive lethal mutations when administered by feeding or injection to adult male Drosophila. Conclusions: Under the conditions of these 2 year feed studies, there was no evidence of carcinogenic activity of tetracycline hydrochloride for male or female F344/N rats and B6C3F1 mice fed diets containing 12,500 or 25,000 ppm. Tetracycline hydrochloride-dosed female rats and male mice had greater survival rates than the respective controls during these studies. Dosed mice had lower body weight than controls, and dosed female mice had no hepatocellular adenomas or carcinomas. Trade Names for Tetracycline or Tetracycline hydrochloride: Achromycin; Amycin; Bristacycline; Cyclopar; Dumocyclin; Neocyclin B; Panmycin; Polycycline; Robitet; Ro-cycline; Steclin; Sumycin; Topicycline; Unimycin PMID- 12724779 TI - Identification of Vangl2 and Scrb1 as planar polarity genes in mammals. AB - In mammals, an example of planar cell polarity (PCP) is the uniform orientation of the hair cell stereociliary bundles within the cochlea. The PCP pathway of Drosophila refers to a conserved signalling pathway that regulates the coordinated orientation of cells or structures within the plane of an epithelium. Here we show that a mutation in Vangl2, a mammalian homologue of the Drosophila PCP gene Strabismus/Van Gogh, results in significant disruptions in the polarization of stereociliary bundles in mouse cochlea as a result of defects in the direction of movement and/or anchoring of the kinocilium within each hair cell. Similar, but less severe, defects are observed in animals containing a mutation in the LAP protein family gene Scrb1 (homologous with Drosophila scribble). Polarization defects in animals heterozygous for Vangl2 and Scrb1 are comparable with Vangl2 homozygotes, demonstrating genetic interactions between these genes in the regulation of PCP in mammals. These results demonstrate a role for the PCP pathway in planar polarization in mammals, and identify Scrb1 as a PCP gene. PMID- 12724780 TI - Association of the T-cell regulatory gene CTLA4 with susceptibility to autoimmune disease. AB - Genes and mechanisms involved in common complex diseases, such as the autoimmune disorders that affect approximately 5% of the population, remain obscure. Here we identify polymorphisms of the cytotoxic T lymphocyte antigen 4 gene (CTLA4)- which encodes a vital negative regulatory molecule of the immune system--as candidates for primary determinants of risk of the common autoimmune disorders Graves' disease, autoimmune hypothyroidism and type 1 diabetes. In humans, disease susceptibility was mapped to a non-coding 6.1 kb 3' region of CTLA4, the common allelic variation of which was correlated with lower messenger RNA levels of the soluble alternative splice form of CTLA4. In the mouse model of type 1 diabetes, susceptibility was also associated with variation in CTLA-4 gene splicing with reduced production of a splice form encoding a molecule lacking the CD80/CD86 ligand-binding domain. Genetic mapping of variants conferring a small disease risk can identify pathways in complex disorders, as exemplified by our discovery of inherited, quantitative alterations of CTLA4 contributing to autoimmune tissue destruction. PMID- 12724782 TI - NTP Toxicology and Carcinogenesis Studies of Benzyl Alcohol (CAS No. 100-51-6) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - Toxicology and carcinogenesis studies of technical-grade benzyl alcohol (99% pure), a textile dye additive, solvent, and food flavoring agent, were conducted by administering the chemical by gavage in corn oil vehicle to groups of F344/N rats and B6C3F1 mice of each sex for 16 days, 13 weeks, or 2 years. Short-Term Studies: In 16-day studies, all five male and five female rats and mice dosed with 2,000 mg/kg benzyl alcohol died. Two of five male and 3/5 female rats and 1/5 male and 2/5 female mice dosed with 1,000 mg/kg died. Rats and mice of each sex in the two highest dose groups were lethargic after dosing. Other toxic responses to benzyl alcohol in these dose groups included blood around the mouth and nose, subcutaneous hemorrhages, and blood in the urinary and gastrointestinal tracts of rats and blood in the urinary bladder of mice. Animals administered lower doses of benzyl alcohol (125, 250, or 500 mg/kg) had no compound-related histologic lesions. Doses selected for the 13-week studies were 0, 50, 100, 200, 400, and 800 mg/kg for rats and mice. Eight of 10 male rats dosed with 800 mg/kg died during weeks 7 and 8; four of these deaths were described as gavage related. Rats dosed with 800 mg/kg exhibited clinical signs indicative of neurotoxicity including staggering, respiratory difficulty, and lethargy. Hemorrhages occurred around the mouth and nose, and there were histologic lesions in the brain, thymus, skeletal muscle, and kidney. In mice, deaths were scattered among all dose levels, but none occurred in vehicle controls. Four male and six female mice died after being dosed; all deaths but one were described as gavage related. Staggering after dosing also occurred during the first 2 weeks of the studies in mice dosed with 800 mg/kg. Some of the deaths in the rats and mice may have been caused by a combination of the gavage procedure and chemical toxicity, since there was evidence that benzyl alcohol induced neurotoxic effects. There were reductions in relative weight gain in male rats dosed with 800 mg/kg benzyl alcohol, in female rats dosed with 200 mg/kg or more, in male mice dosed with 400 or 800 mg/kg, and in female mice dosed with 200 mg/kg or more. No notable changes in body weight gain or compound-related histopathologic lesions were observed in rats or mice from the lower dose groups. Based on mortality, reduction in relative body weight gain, and the histopathologic lesions, doses selected for 2 year studies in rats were 0, 200, and 400 mg/kg. Doses selected for 2-year studies in mice were 0, 100, and 200 mg/kg, based on mortality and depression in relative body weight gain. Body Weight and Survival in the Two-Year Studies: Fifty animals of each species and sex were administered benzyl alcohol in corn oil by gavage 5 days per week for 103 weeks. Administration of benzyl alcohol did not affect survival in male rats (final survival rates: vehicle control, 28/50; low dose, 27/50; high dose, 24/50) but reduced survival of dosed female rats by half (36/50; 18/50; 17/50). Many of the early deaths were considered related to the gavage procedure. Survival in mice was not affected by benzyl alcohol administration (male: 34/50; 33/50; 35/50; female: 26/50; 32/50; 36/50). No effect of benzyl alcohol on body weight gain in rats or mice was observed. In the third month of the studies, clinical signs of sialodacryoadenitis virus infection were observed in rats. A positive serologic reaction for rat coronavirus was observed in sentinel animals at 6 months and again at 18 months. Nonneoplastic and Neoplastic Effects in the Two-Year Studies: No apparent compound-related nonneoplastic responses were observed. Dose-related negative trends in the incidences of anterior pituitary gland neoplasms were seen in female rats (vehicle control, 29/50; low dose, 17/47; high dose, 9/49) and of harderian gland adenomas in male mice (8/50; 3/50; 2/50). Adenomas of the adrenal cortex occurred at an increased incidence in high dose male mice (0/48; 0/44; 3/48), but this slight increase was not considered to be related to chemical expcal exposure. Genetic Toxicology: Benzyl alcohol was not mutagenic when tested by the preincubational protocol in the presence or absence of exogenous metabolic activation in Salmonella typhimurium strains TA98, TA100, TA1535, or TA1537. In the mouse L5178Y/TK± lymphoma assay, benzyl alcohol induced an increase in trifluorothymidine (Tft)-resistant cells in the absence, but not in the presence, of S9; the effect was associated with toxicity. In cytogenetic assays with Chinese hamster ovary (CHO) cells, treatment with benzyl alcohol produced an increase in sister chromatid exchanges (SCEs) which was judged to be equivocal both with and without S9; a significant increase in chromosomal aberrations was observed after exposure to benzyl alcohol in the presence, but not the absence, of S9. Audit: The data, documents, and pathology materials from the 2-year studies of benzyl alcohol have been audited. The audit findings show that the conduct of the studies is documented adequately and support the data and results given in this Technical Report. Conclusions: Under the conditions of these 2-year gavage studies, there was no evidence of carcinogenic activity of benzyl alcohol for male or female F344/N rats dosed with 200 or 400 mg/kg. Survival in both dose groups of female rats was 50% that of vehicle controls, primarily due to an increased number of gavage-related deaths. There was no evidence of carcinogenic activity of benzyl alcohol for male or female B6C3F1 mice dosed with 100 or 200 mg/kg for 2 years. Synonyms: benzenemethanol; phenylcarbinol; phenylmethanol; a-hydroxytoluene; benzenecarbinol; phenolcarbinol; a-toluenol PMID- 12724783 TI - NTP Toxicology and Carcinogenesis Studies of Dichlorvos (CAS No. 62-73-7) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - Toxicology and carcinogenesis studies of dichlorvos (99% pure), a contact and stomach poison for control of insects and parasites, were conducted by administering dichlorvos in corn oil by gavage to groups of F344/N rats and B6C3F1 mice of each sex for 13 weeks or 2 years. Previous feed studies were done by the National Cancer Institute using Osborne-Mendel rats and B6C3F1 mice. Thirteen-Week Studies: Thirteen-week studies with groups of 10 rats of each sex were conducted at doses of 0, 2, 4, 8, 16, 32, or 64 mg/kg dichlorvos in corn oil. All rats that received 32 or 64 mg/kg dichlorvos and 4/10 females that received 16 mg/kg died before the end of the studies. Final mean body weights of dosed and vehicle control rats were similar. Thirteen-week studies with groups of 10 mice of each sex were conducted at doses of 0, 5, 10, 20, 40, 80, or 160 mg/kg. All 10 male mice and 9/10 female mice that received 160 mg/kg and 5/10 male mice that received 80 mg/kg dichlorvos died before the end of the studies. Final mean body weights of dosed and vehicle control mice were similar. No compound-related gross or microscopic pathologic effects were observed in rats or mice. Two-year studies of dichlorvos were conducted by administering 0, 4, or 8 mg/kg dichlorvos, 5 days per week for 103 weeks, to groups of 50 F344/N rats of each sex. Groups of 50 male B6C3F1 mice were administered 0, 10, or 20 mg/kg dichlorvos on the same schedule, and groups of 50 B6C3F1 female mice were administered 0, 20, or 40 mg/kg dichlorvos. Body Weight and Survival in the Two Year Studies: Mean body weights of dosed and vehicle control rats and mice were similar. No significant differences in survival were observed between any groups of rats or mice of either sex (rats--male: vehicle control, 31/50; low dose, 25/50; high dose, 24/50; female: 31/50; 26/50; 26/50; mice-- male: 35/50; 27/50; 29/50; female: 26/50; 29/50; 34/50). Neoplastic Effects in the Two-Year Studies: Adenomas of the exocrine pancreas occurred at greater incidences in dosed rats than in vehicle controls (male: vehicle control, 25/50; low dose, 30/49; high dose, 33/50; female: 2/50; 3/47; 6/50). Mononuclear cell leukemia in both dosed groups of male rats occurred more frequently than in vehicle controls (11/50; 20/50; 21/50). Mammary gland fibroadenomas and fibroadenomas or adenomas (combined) in dosed female rats occurred at increased incidences relative to the vehicle controls (9/50; 19/50; 17/50). Multiple fibroadenomas occurred in dosed female rats but not in vehicle controls (0/50; 6/50; 3/50); carcinomas occurred in two vehicle control and two low dose female rats. In mice, incidences of squamous cell papillomas of the forestomach were increased in the high dose groups compared with those in the vehicle controls (male: 1/50; 1/50; 5/50; female: 5/49; 6/49; 18/50). Two high dose female mice developed forestomach squamous cell carcinomas. Genetic Toxicology: Dichlorvos was mutagenic in Salmonella typhimurium strain TA100 with and without metabolic activation but was not mutagenic in strain TA98. Dichlorvos was mutagenic in the mouse lymphoma L5178Y/TK+/- assay without metabolic activation. Dichlorvos induced sister chromatid exchanges and chromosomal aberrations in Chinese hamster ovary cells in the absence and presence of metabolic activation. Conclusions: Under the conditions of these 2-year gavage studies, there was some evidence of carcinogenic activity of dichlorvos for male F344/N rats, as shown by increased incidences of adenomas of the exocrine pancreas and mononuclear cell leukemia. There was equivocal evidence of carcinogenic activity of dichlorvos for female F344/N rats, as shown by increased incidences of adenomas of the exocrine pancreas and mammary gland fibroadenomas. There was some evidence of carcinogenic activity of dichlorvos for male B6C3F1 mice, as shown by increased incidences of forestomach squamous cell papillomas. There was clear evidence of carcinogenic activity of dichlorvos for female B6C3F1 mice, as shown by increased incidences of forestomach squamous cell papils cell papillomas. Synonyms: 2,2 dichloroethenyl dimethyl phosphate; 2,2-dichlorovinyl dimethyl phosphate; O,O dimethyl-O-(2,2-dichlorovinyl)phosphate; DDVP Trade Names: BAY-19149; DDVF; ENT 20738; OMS-14; SD 1750; Canogard®.; Crossman's Fly-Cake®.; Dedevap®.; De-Pester Insect Strip®.; Estrosol®.; Herkol®.; Kill-fly Resin Strip®.; Lethalaire®.; Mafu®.; Misect®.; Nogos®.; Nuvan®.; No-Pest Strip®.; Oko®.; Phoracide®.; Phosvit®.; Vapona®.; Vaponicide®.; Vaporette Bar®. Anthelmintics: Atgard®.; Dichlorman®.; Equigard®.; Task®. PMID- 12724784 TI - NTP Toxicology and Carcinogenesis Studies of Nitrofurantoin (CAS No. 67-20-9) in F344/N Rats and B6C3F1 Mice (Feed Studies). AB - Nitrofurantoin was studied and evaluated because of its widespread use as a drug for treating urinary tract infections in humans, its structural relationship to known carcinogenic 5-nitrofuran compounds, and the lack of adequate studies to assess its carcinogenicity. Toxicology and carcinogenesis studies of nitrofurantoin were conducted by administering nitrofurantoin (greater than 99% pure) in feed to groups of F344/N rats and B6C3F1 mice of each sex for 14 days, 13 weeks, or 2 years. Fourteen-Day and Thirteen-Week Studies: None of the rats (at dietary concentrations up to 20,000 ppm) died before the end of the 14-day studies. Rats that received 5,000, 10,000, or 20,000 ppm lost weight. Four of five male and 4/5 female mice that received 10,000 ppm and 1/5 females that received 5,000 ppm nitrofurantoin died before the end of the studies. Mice that received 5,000 ppm and male mice that received 10,000 ppm lost weight. In the 13 week studies, final mean body weights of rats that received 2,500, 5,000, or 10,000 ppm were 10%, 34%, or 47% lower than that of the controls for males and 15%, 31%, or 41% lower for females. Feed consumption by dosed and control rats was generally similar. Degeneration of the germinal epithelium of the seminiferous tubules of the testis was observed in male rats that received 2,500 to 10,000 ppm nitrofurantoin. Necrosis of the ovarian follicles was observed in 8/10 female rats that received 10,000 ppm, in 3/10 females that received 5,000 ppm, and in 1/10 that received 2,500 ppm. For mice, final mean body weights of the 5,000-ppm groups were 13% lower than that of the controls for males and 15% lower for females. Two of 10 male mice that received 5,000 ppm and 1/10 males that received 300 ppm died before the end of the 13-week studies. Estimated feed consumption was similar for dosed and control groups. Degeneration of the germinal epithelium of the testis was observed in males that received 1,300 to 5,000 ppm; necrosis of the ovarian follicles was observed in females that received 5,000 ppm but not in the lower dose groups. Necrosis of the renal tubular epithelium was observed in 2/9 males that received 5,000 ppm. Based on these results, 2-year studies of nitrofurantoin were conducted by feeding diets containing 0, 1,300, or 2,500 ppm nitrofurantoin to groups of 50 male F344/N rats and to groups of 50 male and female B6C3F1 mice for 103 weeks. Groups of 50 female F344/N rats were fed diets containing 0, 600, or 1,300 ppm nitrofurantoin on the same schedule. Body Weight and Survival in the Two-Year Studies: Mean body weight and average daily feed consumption of dosed male and female rats were similar to those of the controls throughout the studies. The average amount of nitrofurantoin consumed per day was estimated to be 60 and 110 mg/kg for low and high dose male rats and 30 and 60 mg/kg for low and high dose female rats. No significant differences in the number of rats surviving to the end of the studies were observed between any groups of rats of either sex (male: control, 24/50; low dose, 27/50; high dose, 26/50; female: 25/50; 26/50; 31/50). Mean body weights of high dose male and female mice were up to 12% lower than those of the controls throughout most of the studies. The average daily feed consumption by dosed mice ranged from 93% to 100% that by controls. The average amount of nitrofurantoin consumed per day was estimated to be 280-300 mg/kg and 570-580 mg/kg for low and high dose mice. The survival of the control group of female mice was lower than that of the dosed groups (control, 19/50; low dose, 37/50; high dose, 37/50). The decrease in survival was most likely related to the increase in microbial infection in the reproductive tract observed in the controls. Groups of male mice had similar survival (28/50; 29/50; 34/50). Nonneoplastic and Neoplastic Effects in the Two-Year Studies: Organs showing toxicity from nitrofurantoin exposure identified in the short-term studies were the testis in male rats and mice, the ovary in female rats and mice, and the kidney in male mice. Lesions oband mice, the ovary in female rats and mice, and the kidney in male mice. Lesions observed in the 2-year studies were in the testis in male rats and mice, ovary in female mice, and kidney in male rats. Chronic nephropathy was observed in nearly all rats, but the severity of the lesions was judged to be greater in dosed male rats. Hyperplasia of the transitional cell epithelium (control, 0/50; low dose, 5/50; high dose, 2/50) and hydronephrosis of the renal pelvis (0/50; 5/50; 2/50) were also observed in dosed male rats. In the standard single sections of the left and right kidney from each rat, tubular cell adenomas were observed in one low dose and two high dose males; a tubular cell carcinoma was observed in another high dose male. Because the number of renal tubular cell neoplasms identified by standard procedures in the dosed male rats was low, additional step sections of the kidney were evaluated. The incidences of tubular cell adenomas derived from the step-sections and original sections (combined) were significantly increased in dosed male rats (adenomas: 3/50; 11/50; 19/50); tubular cell carcinomas occurred in two high dose males only. Lesions considered to be associated with the nephropathy and nitrofurantoin exposure were observed in male rats and included hyperplasia of the parathyroid glands (3/49; 18/47; 23/49), fibrous osteodystrophy of the bone (0/50; 5/50; 5/50), and mineralization of the glandular stomach (1/49; 8/50; 14/50). Atypical cells of the epididymis (0/50; 0/50; 12/50) and degeneration of the testis (0/50; 0/50; 36/50) were observed in high dose male rats. Fibrinoid necrosis of arterioles (1/50; 8/50; 15/50) and perivascular infiltration of mononuclear cells (3/50; 9/50; 19/50) were also observed in the testis of male rats. Interstitial cell adenomas of the testis occurred with a negative trend (47/50; 45/50; 21/50), and no adenomas or carcinomas of the preputial gland were seen in high dose male rats (12/48; 11/50; 0/47). The incidence of clitoral gland neoplasms was increased in low dose female rats (5/44; 10/38; 4/42). Osteosarcomas were observed in the bone of one low dose and two high dose male rats. The historical incidence of osteosarcomas in untreated male F344/N rats is 8/1,937 (0.4%). The incidences of subcutaneous tissue neoplasms in dosed male rats were greater than that in the controls (1/50; 7/50; 5/50). No neoplastic lesions in dosed female rats or male mice were considered to be compound related at the doses of nitrofurantoin administered. For female mice, ovarian atrophy was observed in 48/50 low dose and 49/50 high dose mice but not in controls. Tubular cell adenomas of the ovary (0/50; 0/50; 5/50), benign mixed tumors (tubular and stromal) (0/50; 0/50; 4/50), and granulosa cell tumors (0/50; 3/50; 2/50)) were observed in dosed female mice. One granulosa cell tumor in the high dose group was malignant. Ovarian abscesses (18/50) and suppurative inflammation of the uterus (11/50) were observed in control female mice but not in dosed female mice and are believed to be related to indigenous microbial infections and most likely were the cause of early deaths in this group. Adenocarcinomas of the uterus were seen in one low dose and in one high dose mouse. Testicular aspermatogenesis (1/49; 1/49; 16/50), degeneration of the germinal epithelium (0/49; 3/49; 23/50), and atypical cells (0/50; 0/49; 26/50) and depletion (1/50; 1/49; 15/50) of the epididymis were observed at increased incidences in high dose male mice. Spindle cell hyperplasia of the adrenal cortex was observed in dosed female mice (3/50; 41/50; 45/50). A spindle cell adenoma (adrenal capsule adenoma) was seen in one low dose female mouse, and a spindle cell carcinoma (adrenal capsule carcinoma) was seen in one low dose male mouse. Mineralization of the renal medulla (male: 0/50; 0/50; 17/50; female: 0/50; 0/50; 7/50) and dilatation of the renal tubules (male: 0/50; 0/50; 14/50) were observed in high dose mice. Hepatocellular neoplasms (adenomas or carcinomas, combined) were observed at an increased incidence in high dose female mice (2/50; 2/50; 8/50). An Ito cell tumor of the liver was observed in one low dose and one high dose female mouse. Malignant lymphomas occurred in female mice (12/50; 19/50; 24/50). Genetic Toxicology: Nitrofurantoin was mutagenic in Salmonella typhimurium strains TA98 and TA100, with and without metabolic activation, but was not mutagenic for strains TA1535 or TA1537. Nitrofurantoin induced forward mutations at the TK+/- locus of L5178Y mouse lymphoma cells in the absence of metabolic activation (it was not tested with activation). Nitrofurantoin induced increased numbers of sister chromatid exchanges and chromosomal aberrations in cultured Chinese hamster ovary cells with and without metabolic activation. Results of the sex-linked recessive lethal assay in Drosophila were negative after administration of nitrofurantoin by feeding or by injection. Conclusions: Under the conditions of these 2-year feed studies, there was some evidence of carcinogenic activity of nitrofurantoin for male F344/N rats as shown by increased incidences of uncommon kidney tubular cell neoplasms. Uncommon osteosarcomas of the bone and neoplasms of the subcutaneous tissue were observed in dosed male rats. Incidences of interstitial cell adenomas of the testis and neoplasms of the preputial gland were decreased in the 2,500-ppm group of male rats. There was no evidence of carcinogenic activity of nitrofurantoin for female F344/N rats fed diets containing 600 ppm or 1,300 ppm for 2 years. Female rats may have been able to tolerate higher doses. There was no evidence of carcinogenic activity of nitrofurantoin for male B6C3F1 mice fed diets containing 1,300 ppm or 2,500 ppm for 2 years. There was clear evidence of carcinogenic activity of nitrofurantoin for female B6C3F1 mice as shown by increased incidences of tubular adenomas, benign mixed tumors, and granulosa cell tumors of the ovary. Nonneoplastic lesions considered related to nitrofurantoin exposure were chronic nephropathy and associated lesions (hyperplasia of the parathyroid gland, fibrous osteodystrophy of the bone, and mineralization of the glandular stomach) in male rats and testicular degeneration in male rats and mice. Ovarian atrophy and hyperplasia of the adrenal cortex spindle cells were observed in dosed female mice. Synonyms: 1-(((5-nitro-2-furanyl)methylene)amino-2,4 imidazolidinedione); 1-(5-nitro-2-furfurylideneamino)-hydantoin; N-(5-nitro-2 furfurlidene)-1-aminohydantoin; 1-((5-nitrofurfurylidene)amino)hydantoin Trade Names: Benkfuran; Benkfurin; Chemiofuran; Cyantin; Dantafur; Furadantin; Furadantine; Furadantoin; Furadonin; Furadonine; Furantoin; Furatoin; Furobactina; Ituran; Macrodantin; Nifurantin; NSC 2107; N-Toin; Orafuran; Parafuran; Urizept; USAF EA-2; Welfurin; Zoofurin PMID- 12724785 TI - NTP Toxicology and Carcinogenesis Studies of Iodinated Glycerol (Organidin(R).) (CAS No. 5634-39-9) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - Toxicology and carcinogenesis studies of iodinated glycerol (Organidin(R)., a complex mixture prepared by the reaction of iodine with glycerol and found to contain 33% 3-iodo-1,2-propanediol as the major component) were conducted because of human exposure to iodinated glycerol as an expectorant and its possible relationship to the formation of alkyl iodides, e.g., methyl iodide, a suspected animal carcinogen. These studies were conducted by giving iodinated glycerol in water by gavage (5 days per week) to groups of F344/N rats and B6C3F1 mice for 16 days, 13 weeks, or 2 years. Genetic toxicology studies were conducted with iodinated glycerol in Salmonella typhimurium, mouse L5178Y lymphoma cells, Chinese hamster ovary (CHO) cells, and B6C3F1 mice (in vivo bone marrow micronucleus test). Also, 3-iodo-1,2-propanediol was tested in S. typhimurium and B6C3F1 mice (in vivo micronucleus assay). Sixteen-Day and Thirteen-Week Studies: Sixteen-day studies were conducted by giving iodinated glycerol at doses up to 1,000 mg/kg to rats and up to 500 mg/kg to mice. All female rats and 4/5 male mice in the highest dose group died before the end of the studies; there were no dose-related effects on body weights of male or female rats or male mice at the end of the studies. The forestomach of 2/5 female mice that received 500 mg/kg was thickened and granular. Thirteen-week studies were conducted by administering iodinated glycerol at doses up to 500 mg/kg to rats and mice. During these studies, 3/10 female rats and 1/10 female mice that received 500 mg/kg died. Final mean body weights of rats and mice that received 500 mg/kg were 4% lower than those of vehicle controls for males and 6%-7% lower for females. Kidney tubular cell lesions, including cortical necrosis, regeneration, and calcification, were observed at increased incidences in the highest dose group of female rats. Lymphoid hyperplasia of the stomach was observed in dosed male and female rats. Kidney tubular cell regeneration was also observed in dosed female mice. Inflammation or abscesses of mild-to-moderate severity and hyperplasia, acanthosis, and/or hyperkeratosis of mild-to-moderate severity were observed in the forestomach of the highest dosed group of female mice. Body Weight and Survival in the Two-Year Studies: Two-year studies were conducted by administering 0, 125, or 250 mg/kg iodinated glycerol in deionized water by gavage, 5 days per week for 103 weeks, to groups of 50 male F344/N rats and 50 male B6C3F1 mice. Groups of 50 female F344/N rats and 50 female B6C3F1 mice were administered iodinated glycerol on the same schedule at lower doses of 0, 62, or 125 mg/kg because of the increased severity of kidney and stomach lesions in the 13-week studies. Mean body weights of high dose male rats were 5%-10% lower than those of vehicle controls from week 43 to week 68 and 10%-13% lower from week 72 to the end of the studies. Mean body weights of low dose male rats and high dose female rats were 4%-9% lower than those of vehicle controls from week 88 to the end of the studies. The survival of the high dose group of male rats was considerably lower than that of the vehicle controls after week 86. No other significant differences in survival were observed between any groups of rats of either sex (male: vehicle control, 28/50; low dose, 20/50, high dose, 2/50; female: 31/50; 30/50; 27/50). Mean body weights of dosed and vehicle control male mice were similar. Mean body weights of high dose female mice were 6%-8% lower than those of vehicle controls from week 40 to week 64 and were 9%-13% lower thereafter. No significant differences in survival were observed between any groups of mice of either sex (male: 36/50; 40/50; 32/50; female: 40/50; 33/50; 38/50). Nonneoplastic and Neoplastic Effects in the Two-Year Studies: The incidence of mononuclear cell leukemia were increased in dosed male rats (vehicle control, 14/50; low dose, 29/50; high dose, 24/50). Follicular cell carcinomas of the thyroid gland in male rats occurred at an increased incidence in low dose male rats (0/49; s of the thyroid gland in male rats occurred at an increased incidence in low dose male rats (0/49; 5/49; 1/49). Reduced survival of high dose male rats may have been responsible for the decreased tumor incidence in this group relative to that in the low dose group. Follicular cell carcinomas were observed in one low dose and one high dose female rat. Follicular cell carcinomas of the thyroid gland have been observed in 3/293 water gavage vehicle control male F344/N rats and in 10/1,904 untreated control male F344/N rats. Adenomas of the nasal cavity were observed in two high dose male rats. Adenomas of the nasal cavity have not been observed in 300 water gavage vehicle control male F344/N rats or in 1,936 untreated control male F344/N rats. Squamous metaplasia and focal atrophy of the salivary glands were observed at increased incidences in dosed rats (squamous metaplasia--male: 0/48; 47/50; 48/49; female:1/49; 48/50; 49/50; focal atrophy--male: 1/48; 10/50; 30/49; female: 0/49; 4/50; 11/50). In dosed female mice, adenomas of the anterior pituitary gland were increased (10/47; 15/45; 24/46). The incidences of adenomas of the harderian gland in dosed female mice were increased (6/50; 8/40; 13/50). A carcinoma of the harderian gland was observed in another high dose female mouse. Dilatation of the thyroid gland follicle and follicular cell hyperplasia were observed at increased incidences in dosed mice (dilatation--male: 0/48; 28/50; 32/50; female: 4/48; 11/48; 10/48; hyperplasia--male: 3/48; 46/50; 34/50; female: 2/48; 25/48; 35/48). The incidences of follicular cell adenomas were 3/48, 6/50, and 0/50 for males and 2/48, 3/48, and 4/48 for females. Hyperkeratosis and acanthosis of the forestomach were observed at increased incidences in high dose male mice (hyperkeratosis: 0/49; 0/49; 5/50; acanthosis: 0/49; 1/49; 5/50). Squamous cell papillomas were observed in female mice (1/49; 2/50; 5/49). The historical incidence of forestomach squamous cell neoplasms is 4/339 (1.2%) in water gavage vehicle control female B6C3F1 mice and is 18/1,994 (0.9%) in untreated control female B6C3F1 mice. Squamous cell neoplasms were not observed in male mice. Genetic Toxicology: Treatment of the base-substitution mutant S. typhimurium strains TA100 and TA1535 with iodinated glycerol in a preincubational protocol with and without S9 resulted in a dose-related increase in the number of revertant colonies; no increase in revertants was observed with the frame-shift mutant strains TA98 or TA1537. 3-Iodo-1,2-propanediol was also mutagenic in TA100 with or without S9; it was not mutagenic in TA98. Iodinated glycerol increased the number of trifluorothymidine-resistant cells in mouse lymphoma L5178Y/TK± assay in the absence of exogenous metabolic activation; it was not tested with activation. Iodinated glycerol induced sister chromatid exchanges (SCEs) and chromosomal aberrations in CHO cells without S9; with S9, the frequency of SCEs was increased more than without S9 but no chromosomal aberrations were induced. No increase in micronucleated polychromatic erythrocytes was observed in the bone marrow of B6C3F1 mice after injection with either iodinated glycerol or 3-iodo-1,2-propanediol. Conclusions: Under the conditions of these 2-year gavage studies, there was some evidence of carcinogenic activity for male F344/N rats administered iodinated glycerol, as indicated by increased incidences of mononuclear cell leukemia and follicular cell carcinomas of the thyroid gland. Adenomas of the nasal cavity in two high dose male rats may have been related to the administration of iodinated glycerol. There was no evidence of carcinogenic activity for female F344/N rats administered 62 or 125 mg/kg iodinated glycerol by gavage for 103 weeks. There was no evidence of carcinogenic activity for male B6C3F1 mice administered 125 or 250 mg/kg iodinated glycerol by gavage for 103 weeks. There was some evidence of carcinogenic activity for female B6C3F1 mice administered iodinated glycerol, as indicated by increased incidences of adenomas of the anterior pituitary gland and neoplasms of the harderian gland. Squamous cell papillomas of the forestomach may have been related to the administration of iodinated glycerol. Significant nonneoplastic lesions considered related to exposure of iodinated glycerol were squamous metaplasia and focal atrophy of the salivary gland in male and female rats. Dilatation of the thyroid gland follicle and follicular cell hyperplasia were observed in male and female mice. Synonyms or Trade Names: Organidin®.; iodopropylidene glycerol PMID- 12724786 TI - NTP Toxicology and Carcinogenesis Studies of 2-Amino-4-Nitrophenol (CAS No. 99-57 0) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - 2-Amino-4-nitrophenol is used to color semipermanent hair dyes and in the manufacture of mordant dyes for leather, nylon, silk, wool, and fur. 2-Amino-4 nitrophenol was nominated by the National Cancer Institute for toxicology and carcinogenesis studies because of widespread human exposure associated with its manufacture and use. Toxicology and carcinogenesis studies were conducted by administering 2-amino-4-nitrophenol (98% pure) in corn oil by gavage, 5 days per week, to groups of F344/N rats and B6C3F1 mice of each sex in 15-day, 13-week, and 2-year studies. Fifteen-Day and Thirteen-Week Studies: During the 15-day studies, rats and mice received doses of 0, 313, 625, 1,250, 2,500, or 5,000 mg/kg. All rats that received 2,500 or 5,000 mg/kg and all female rats that received 1,250 mg/kg died before the end of the studies. Final mean body weights of chemically exposed rats surviving to the end of the studies were comparable to those of vehicle controls. Diarrhea was observed in all groups of exposed rats except those receiving 313 mg/kg. All mice that received 2,500 or 5,000 mg/kg, 2/5 males and all females that received 1,250 mg/kg, and 1/5 females that received 313 mg/kg died before the end of the studies. Final mean body weights of exposed mice surviving until the end of the studies were comparable to those of vehicle controls. In 13-week studies, F344/N rats and B6C3F1 mice of each sex received 2-amino-4-nitrophenol at doses of 0, 62.5, 125, 250, 500, or 1,000 mg/kg. All rats that received 1,000 mg/kg and 2/10 males and 2/10 females that received 500 mg/kg died before the end of the studies. The final mean body weight of male rats that received 500 mg/kg was reduced 10% compared with that of vehicle controls; final mean body weights of all other surviving exposed rat groups were comparable to those of vehicle controls. Diarrhea and lethargy were observed for rats that received 500 or 1,000 mg/kg. All male mice and most females that received 1,000 mg/kg and 4/10 females that received 500 mg/kg died before the end of the studies. Final mean body weights of chemically exposed mice were comparable to those of vehicle controls. No compound-related clinical signs were observed in mice during the studies. Mineralization of the renal cortex and degeneration of the renal tubular epithelium were observed in male and female rats that received 1,000 mg/kg and in males that received 500 mg/kg. Degeneration and necrosis of the renal tubular epithelium was observed in 5/10 male and 3/10 female mice that received 1,000 mg/kg. Body Weight and Survival in the Two-Year Studies: In the 2-year studies, rats and mice received 2-amino- 4- nitrophenol at doses of 0, 125, or 250 mg/kg. Mean body weights of male rats that received 250 mg/kg were 8%-10% lower than those of vehicle controls throughout most of the 2 year study. Mean body weights of female rats were comparable to those of vehicle controls. Soft stools and occasional diarrhea were observed in chemically exposed rats starting 6 months after the beginning of the studies. Survival of male rats that received 250 mg/kg was markedly lower than that of vehicle controls after week 89 (final survival: vehicle control, 32/50; 125 mg/kg group, 24/50; 250 mg/kg group, 10/50). Survival of female rats was comparable among all groups (final survival: 25/50; 27/50; 31/50). Mean body weights of male and female mice that received 250 mg/kg were comparable to those of vehicle controls; the mean body weights of female mice that received 125 mg/kg were as much as 17% greater than that of vehicle controls. Survival of all mouse groups was comparable during the 2-year studies (final survival: male-- 28/50; 29/50; 23/50; female--28/50; 31/50; 30/50). Nonneoplastic and Neoplastic Effects in the Two-Year Studies: Pigmentation of the small and large intestines was present in exposed rats but not in vehicle controls. Ulcers and erosive lesions of the digestive tract were observed in male rats that received 250 mg/kg and to a lesser extent in male rats that received 125 mg/kg. A carcinoma of the colon occurrkg. A carcinoma of the colon occurred in one male rat that received 250 mg/kg; no other neoplasms were observed in the gastrointestinal tract of rats. No pigmentation, ulcers, or erosive lesions were found in the digestive tract of mice. The severity of nephropathy was markedly greater in exposed male rats than in vehicle controls. Associated with the nephropathy were nonneoplastic lesions indicative of reduced renal function and secondary hyperparathyroidism, including parathyroid hyperplasia, mineralization of various organs, and fibrous osteodystrophy. Renal tubular cell hyperplasia (1/50; 4/48; 5/50) and renal cortical (tubular cell) adenomas (0/50; 1/48; 3/50) occurred in male rats. Renal cortical adenomas are infrequently observed in male F344/N rats (historical incidence, 0.5%). More preputial gland adenomas or carcinomas (combined) were observed in low dose male rats than in vehicle controls (3/50; 10/48; 3/50), whereas the incidences of clitoral gland neoplasms were decreased in dosed female rats (9/50; 6/50; 1/49). Hemangiomas or hemangiosarcomas (combined) occurred in male mice that received 2 amino-4-nitrophenol (0/50; 1/50; 5/50); each tumor was present at a different site. The historical control incidence is 11% at the study laboratory and 6% in 2-year NTP studies. Genetic Toxicology: 2-Amino-4-nitrophenol was mutagenic in Salmonella typhimurium strains TA98 and TA100 with metabolic activation. 2-Amino-4-nitrophenol was not mutagenic in strains TA1535 or TA1537. 2-Amino-4-nitrophenol was mutagenic in the mouse lymphoma L5178Y/TK± assay without metabolic activation. It was not tested with activation. 2-Amino-4 nitrophenol induced sister chromatid exchanges (SCEs) and chromosomal aberrations in Chinese hamster ovary cells in the presence and absence of metabolic activation. Audit: The data, documents, and pathology materials from the 2-year studies of 2-amino-4-nitrophenol were audited at the NTP Archives. The audit findings show that the conduct of the studies is documented adequately and support the data and results given in this Technical Report. Conclusions: Under the conditions of these 2-year gavage studies, there was some evidence of carcinogenic activity of 2-amino-4-nitrophenol for male F344/N rats, as shown by increased incidences of renal cortical (tubular cell) adenomas. The incidences of renal tubular cell hyperplasia were also increased in male rats exposed to 2 amino-4-nitrophenol. The survival of male rats that received 2-amino-4 nitrophenol was reduced compared with survival of vehicle control male rats. There was no evidence of carcinogenic activity of 2-amino-4-nitrophenol for female F344/N rats or for male or female B6C3F1 mice that received 125 or 250 mg/kg per day. PMID- 12724787 TI - NTP Toxicology and Carcinogenesis Studies of Roxarsone (CAS No. 121-19-7) in F344/N Rats and B6C3F1 Mice (Feed Studies). AB - Roxarsone is a veterinary drug used as a growth promoter and as an anticoccidial agent and for treatment of swine dysentery. Toxicology and carcinogenesis studies were conducted by administering roxarsone (greater than 99.4% pure) in feed to groups of F344/N rats and B6C3F1 mice of each sex for 14 days, 13 weeks, or 2 years. Fourteen-Day and Thirteen-Week Studies: In the 14-day studies, the diets fed to rats contained 0 or 100-1,600 ppm roxarsone, and those fed to mice contained 0 or 60-1,000 ppm. Deaths occurred in rats and mice that received the highest doses. Rats that received 800 or 1,600 ppm lost weight. Male mice that received 1,000 ppm and female mice that received 500 ppm lost weight. In the first 13-week studies, roxarsone was fed to rats and mice at dietary concentrations of 0 or 50-800 ppm. Decreases (more than 10%) in final mean body weights of dosed rats relative to those of controls were observed for males that received 200, 400, or 800 ppm and for females that received 400 or 800 ppm. Deaths occurred in groups that received 800 ppm. Clinical signs of toxicity (trembling, ataxia, and pale skin) were seen primarily in rats that received 800 ppm. Kidney lesions were observed in rats that received 800 ppm. These lesions were characterized by tubular necrosis and mineralization in the rats that died during the studies and by tubular dilatation and casts, interstitial inflammation, and tubular epithelial cell regeneration in the rats that lived to the end of the studies. Additional 13-week studies were conducted in rats at dietary concentrations of 0, 100, or 400 ppm to demonstrate the absorption of roxarsone from the gastrointestinal tract; to determine its distribution in liver, kidney, and blood; and to study its effects on various hematologic and clinical chemical values. No deaths occurred. Renal lesions of minimal severity observed in male rats that received 400 ppm were characterized by tubular epithelial cell degeneration and regeneration, tubular casts, and mineralization. Arsenic levels in urine, blood, kidney, and liver of dosed rats increased (140% 300%) with time on study and were proportional to the dietary concentrations of roxarsone. No compound-related hematologic or clinical chemical effects were observed in rats. In the first 13-week studies, final mean body weights of mice that received 800 ppm were 11%-18% lower than those of controls. Deaths occurred in males and females receiving 400 and 800 ppm. No compound-related gross or histopathologic lesions were observed. In the second 13-week studies in mice, no compound-related hematologic or clinical chemical effects were observed. At the end of the studies, arsenic concentrations in dosed mice ranged from 0.45 to 0.99 ug/g of liver and from 0.85 to 2.98 ug/g of kidney. No arsenic was detected in the liver or kidney of control mice. Because of kidney lesions, lower body weight gain, and increased mortality in rats and lower body weight gain and increased mortality in mice in the short-term studies, dietary concentrations of roxarsone selected for the 2-year studies were 0, 50, or 100 ppm for rats and 0, 100, or 200 ppm for mice. Body Weight and Survival in the Two-Year Studies: Mean body weights of dosed rats were generally within 5% of those of controls. No significant differences in survival were observed between any groups of rats of either sex, although survival in males was lower than usual (final survival- male: control, 24/50; low dose, 18/50; high dose, 18/50; female: 27/50; 35/50; 32/50). The average feed consumption by high dose rats was 95% that of controls for males and 88% for females. The average amount of roxarsone consumed per day was approximately 2 mg/kg for low dose rats and 4 mg/kg for high dose rats. Mean body weights of high dose male mice were generally 5%-8% higher than those of the controls, whereas those of female mice were generally 6%-15% lower than those of the controls. The survival of the control group of male mice was lower than that of the low dose group after month 22; survival for females was low (final survir than that of the low dose group after month 22; survival for females was low (final survival--male: 27/50; 40/50; 33/50; female: 14/50; 18/50; 17/50). The low survival in females was due in part to utero-ovarian infection, with more than 50% of the animals in each dose group having suppurative inflammation at this site. The average daily feed consumption by dosed mice was 105% 110% that by the controls. The average amount of roxarsone consumed per day was approximately 21 or 43 mg/kg for low dose or high dose male mice and 27 or 54 mg/kg for low dose or high dose female mice. Nonneoplastic and Neoplastic Effects in the Two-Year Studies: Although the incidence of adenomas of the exocrine pancreas in high dose male rats was not statistically greater than that in the controls (control, 1/50; low dose, 1/50; high dose, 5/50), it was greater than that seen in any historical control group of male F344/N rats. The historical rate is 1/437 (0.2%) for the study laboratory and 5/1,871 (0.3%) throughout the Program. The incidences of hyperplasia were 2/50; 0/50; 3/50. No hyperplasia oradenomas were observed in the exocrine pancreas of female rats. Clitoral gland adenomas in female rats occurred with a marginally positive trend (1/44; 3/47; 6/48; P=0.049). One carcinoma was also observed in each of the groups. The incidences of adenomas or of adenomas or carcinomas (combined) in the dosed groups were not significantly different from those in the controls. This marginal effect was not considered to be related to roxarsone administration. No chemical-related increases in neoplastic or nonneoplastic lesions occurred in male or female mice. Lymphomas in female mice occurred with a negative trend; the incidences in the dosed groups were lower than that in the controls (13/50; 2/50; 3/50; P≤0.01). Genetic Toxicology: Roxarsone was not mutagenic in Salmonella typhimurium strains TA98, TA100, TA1535, or TA1537 with or without metabolic activation. Roxarsone induced trifluorothymidine (Tft) resistance in mouse lymphoma L5178Y cells in the absence of metabolic activation; it was not tested with activation. Exposure of adult male Drosophila melanogaster to roxarsone by injection or by feeding did not cause an increase in sex--linked recessive lethal mutations. Audit: The data, documents, and pathology materials from the 2-year studies of roxarsone have been audited. The audit findings show that the conduct of the studies is documented adequately and support the data and results given in this Technical Report. Conclusions: Under the conditions of these 2-year feed studies, there was equivocal evidence of carcinogenic activity of roxarsone for male F344/N rats, as indicated by a marginally increased incidence of adenomas of the exocrine pancreas. There was no evidence of carcinogenic activity for female F344/N rats fed diets containing 50 or 100 ppm roxarsone for 2 years. There was no evidence of carcinogenic activity for male or female B6C3F1 mice fed diets containing 100 or 200 ppm roxarsone for 2 years. Synonyms: 4-hydroxy-3-nitrophenylarsonic acid; 4-hydroxy-3-nitrobenzenearsonic acid; 2-nitro-1-hydroxybenzene-4-arsonic acid; nitrophenolarsonic acid; 3-nitro-4 hydroxybenzenearsonic acid; 3-nitro-4-hydroxyphenylarsonic acid Trade Names: Ristat; Ren-O-sal; 3-nitro; 3-nitro-10; 3-nitro-20; 3-nitro-50; 3-nitro-80 PMID- 12724788 TI - [Organ transplantation in Brazil]. PMID- 12724793 TI - [Neovagina: progressive dilatation or surgery?]. PMID- 12724804 TI - [Retirement for permanent disability]. PMID- 12724803 TI - [Anti-mullerian hormone: importance in pediatric practice]. PMID- 12724806 TI - [Sepsis by sinusopathy in an immunodepressed patient]. PMID- 12724807 TI - [Endogenous suicide in plastic surgery]. PMID- 12724808 TI - [Increased skin pigmentation in patients with chronic renal failure undergoing hemodialysis infected with the hepatitis C virus]. AB - BACKGROUND: Skin pigmentation is commonly found in patients with chronic renal failure (CRF). This symptom is also one of the most evident features of Porphyria Cutanea Tarda (PCT). Hepatitis C virus (HCV) is an important precipitating agent of this disease, which has been described in patients undergoing hemodialysis (HD). In this paper, we intend to evaluate the prevalence of difuse skin hyperpigmentation in patients with CRF infected with the HCV. METHODS: We developed a transversal study with 47 patients (mean age 50,35 + 15,16 years; 31 men and 16 women) who were on hemodialysis in march of 2001 at the Unit of Dialysis of Hospital de Cl nicas de Porto Alegre. Patients were divided in two groups: group 1, anti-HCV positives (n=17), and group 2, anti-HCV negatives (n=30). RESULTS: The prevalence of skin hyperpigmentation related to HD was 36,2% (n=17). There were 10 patients (58,8%) in group 1 and 7 (23,3%) in group 2 (Odds Ratio of 2,52 and 95% Confidence Interval of 1,18-5,4; p<0,05). Among patients who have undergone HD for until 36 months (n=24), 6 were anti-HCV positive and 3 of tham (50%) mentioned skin pigmentation (OR of 9,0 and 95%CI of 1,1-71,0; p<0,05). Patients from group 2 who referenced the symptom have been on HD for a longer period than those who did not (63,85 + 11,9 vs. 29,3 + 4,71 months; p<0,05). CONCLUSIONS: HCV infection was associated with increased skin hyperpigmentation in patients with CRF undergoing HD. Time on treatment was also associated with this signal. PMID- 12724809 TI - [Early diagnosis of childhood cancer: a team responsibility]. AB - OBJECTIVES: To call attention for necessity of the knowledge of the early symptoms of childhood cancer and for the responsibility of the parents, physicians and teachers rendering health supervision to the children and to alert that the early detection and prompt treatment is of paramount importance in achieving cures in childhood cancer. SOURCES: Literature review using Medline. SUMMARY: The diagnosis of childhood cancer is complex and many variables play an important role. Early detection and prompt therapy have the potential to reduce mortality and treatment morbidity. A better understanding of the relationship between symptoms and diagnosis is important for the development of effective public health strategies. CONCLUSIONS: There are several factors that may be associated with length of time between symptoms and diagnosis. Lag time to diagnosis depends on tumor type and age. To determine warning signs and symptoms that should alert to the possibility of malignant disease is still a challenge. PMID- 12724810 TI - [The phagocytosis by polymorphonuclear neutrophils in patients with systemic lupus erythematosus]. AB - OBJECTIVE: To evaluate the presence of immune complexes and the phagocytes by polymorphonuclear neutrophils in patients with systemic lupus erythematosus, with and without disease activity. METHODS: The peripheral blood of 55 subjects was analyzed. Ten of those subjects had disease activity, 15 had not disease activity, and 30 were healthy. We used radial immune diffusion to detect immune complexes. The phagocytic function was estimated by the ingestion of zymosan by polymorphonuclear neutrophils. RESULTS: In this study we found the presence of immune complexes formatted of IgM, IgG, IgA, and complement component C3 and C4 in LES patients. The arithmetic average of zymosan particles ingested by the neutrophils incubated with homologous human serum and autologous human serum was significantly decreased (p<0.05) in the LES activity patients when we compare with the group without activity, and the control group. CONCLUSION: We conclude that there are immune complexes in the LES patients with and without disease activity, and there is a reduction in the digestive step of the phagocytes by polymorphonuclear neutrophils in patients with disease activity. The conclusions of the present study are according with the pathogenesis of the disease and with the high mortality in these patients. PMID- 12724811 TI - [Serum and salivary immunoglobulin A levels in patients with cancer of the mouth and oropharynx]. AB - OBJECTIVE: Patients carriers of head and neck cancer (HNC) may show changes in concentrations of serum and salivary IgA owing to an inespecific immunologic disorder that follows the development of malignant lesions. PURPOSE: Evaluate the serum and salivary IgA levelS in Patients With Hnc. METHODS: A prospective study based on a sample of 34 patients with squamous cell carcinoma of the mouth and oropharynx and 34 normal control cases, matched by sex and age. Blood and saliva samples were collected at the same time and assayed for IgA by nephelometry and single radial immunodiffusion (RID). Statistical analysis included Student t Test, ANOVA and Pearson correlation index. RESULTS: The differences between nephelometry and RID could not be detected (p=0.039). The serum concentrations of IgA were 279.4 +/- 131.7 mg/dl and 310.9 +/- 194.1 mg/dl for control and study groups, respectively. Concerning salivary IgA, levels obtained by nephelometry were 17.0 +/- 10.4 mg/dl for control cases and 7.2 +/- 5.0 mg/dl for cancer cases and RID showed concentrations of 13.7 9.1 mg/dl and 5.6 +/- 4.2 mg/dl for control and study group, respectively. There were no significant correlations between serum or salivary IgA levels and age or disease stage. CONCLUSION: Patients carriers of HNC and control subjects showed similar serum concentrations of IgA but it was found that salivary IgA levels were reduced in cancer patients. Causes associated with decreased salivary IgA levels like malnutrition, stress and tobacco could be related to these findings. PMID- 12724812 TI - [Prevalence of immunohematologic tests at birth and the incidence of hemolytic disease in the newborn]. AB - The administration of anti-D globulin to the mothers has decreased the incidence of Rh hemolytic disease but the improvement of technologic assays has made it possible to identify several hemolytics diseases of the newborn. BACKGROUND: To identify the prevalence of immunohematologic tests demonstrated by indirect (IC), direct (DC) and elution tests; to identify the incidence of hemolytic disease and its treatment (phototherapy and/or exchange transfusion) in neonates with hemolytic disease. This is a retrospective cohort-study performed from January 1st 1996 to July 1st 1998. METHODS: This is a descriptive study of the immunohematologic profile of 1698 mothers and their offsprings, as risk factors for developing hemolytic disease. The inclusion criteria were the positivity of the indirect (IC) and direct (DC) Coombs tests and elution tests. Based on the inclusion criteria three group of infants were analyzed: Group I was composed of 149 offsprings of Coombs-positive mothers (IC+) with antibodies associated with neonatal hemolytic disease. This group was further divided into two groups: Group I-A (IC+DC+) was composed of 83 Coombs-positive offsprings (DC+) of Coombs positive mothers (IC+) and Group I-B (IC+DC-) was composed of 66 Coombs-negative offsprings (DC-) of Coombs-positive mothers (IC+); Group D was composed of 736 Coombs-positive offspring's of Coombs-negative mothers (IC-); and Group E was composed of 807 Coombs-negative and elution-positive offspring's of Coombs negative mothers. RESULTS: This study shown that the overall prevalence of immunohematologic tests associated with hemolytic disease was 9.07% (3212/35429), 0.43% (154/35429) among offsprings of Coombs-positive mothers, 4.10% (1453/35429) among Coombs-positive infants, and 4.53% (1605/35429) among elution-positive infants. The overall incidence of hemolytic disease was 36.23% (613/1692), 33.56% (50/149) among offspring's of Coombs-positive mothers, 44.43% (327/736) among Coombs-positive infants, and 29.24% (236/807) among elution-positive infants. The overall incidence of phototherapy among infants with hemolytic disease was 36.23% (613/1692), 49.40% (41/83) in group I-A (IC+DC+), 13.64% (9/66) in group I B(IC+DC-), 44.43% (327/736) in group D, and 29.24% (236/807) in group E. The overall incidence of exchange transfusion among infants with hemolytic disease was 0.88% (15/1692), 14.46% (12/83) in group I-A (IC+DC+), 0% (0/66) in group I-B (IC+DC-), 0.27% (2/736) in group D, and 0.12% (1/807) in group E. CONCLUSIONS: The results of this study allowed us to conclude that the overall prevalence of immunohematologic tests associated with hemolytic disease was 9.07% (3212/35429) and the overall incidence of hemolytic disease was 36.23% (613/1692) in this study-group. The highest incidences of hemolytic disease and phototherapy were observed among Coombs-positive offsprings of Coombs-positive mothers. PMID- 12724813 TI - [Evaluation of condylar fractures treatment]. AB - BACKGROUND: We analysed 40 patients with mandibular condylar process fractures, treated through the closed (intermaxilary fixation or conservatively) or open methods (wire internal fixation,titanium miniplates, bone reduction or articular exploration), from January 1993 to January 1999. METHODS: We evaluated these patients clinically with pre-determined criteria through the temporomandibular articulation touching (ATM), by observing the mandibular symmetry, dental oclusion, deviation to the mouth opening and maximum interincisal opening, besides the questionnaire about the complaints at ATM, disfunction of the mastication system and satisfaction of the treatment. We found an index of good clinical results of, at least, 70% (in the maximum interincisal opening) in the patients submitted to IMF. In the conservative method, we found an index of, at least, 42.9% and in the patients submitted to wire open reduction, at least, 28.6% (in the ATM touching exam) without significant difference in the clinical exams; the pain index at ATM in the closed method was 3.7% and in the open one was 15.4%; mastication disfunction was 22.2% in the closed method and 15.4% in the open one; we only found 7.1% dissatisfied patients in the closed method. There was no significant difference in the method results. CONCLUSIONS: The closed method - with IMF and conservative - and the open reduction - with ORIF - brought good clinical results in an evaluation up to 6 months. We could also conclude that there is no difference in the complaint results after the open or closed treatment according to the patients' opinion. PMID- 12724814 TI - [Retrospective analysis of permanent disability among retired workers at Universidade Federal de Minas Gerais from 1966 to 1999]. AB - OBJECTIVE: The aim of the present study was to conduct a retrospective analysis of all retirement cases from permanent disability (related or not to work) of the staff members at the Universidade Federal de Minas Gerais (UFMG), from 1966 to 1999. METHODS: a historical series analyses to observational study was done. The considered variables included age, gender, diagnosis, occupation, date of disability leave and data of retirement. RESULTS: It was evidenced from the 355 analysed cases, large variability in the yearly distribution, with minimum variability between the years of 1978/91 and maximum in 1999. From the total retirement cases, 56% were male and 44% female, with a mean age of 51 years. The most frequent diagnoses were schizophrenia (14%), arterial hypertension (13%) and back problems (9%). In relation to occupation, the mean rate of retirement for the teaching group (group 1) was 1.33/1000 workers/year and 1.86/1000 workers/year for the technical and general service workers (group 2). CONCLUSIONS: The findings from the present study associated with the population morbidity data will allow the Servi o de Aten o Sa de do Trabalhador (SAST/Pampulha/UFMG), to better promote health programs in the workplace. PMID- 12724815 TI - [Villitis of unknown etiology in placentas of pregnancies with hypertensive disorders and of small-for-gestational-age infants]. AB - BACKGROUND: The objectives of this study are to analyze the frequency and the histopathological features of the villitis of unknown etiology (VUE) in placentas of pregnancies with hypertensive disorders and of small-for-gestational-age infants (SGA). METHODS: Two hundred and thirteen placentas from pregnancies without clinical or laboratorial evidence of infection were studied. These cases were subdivided according to: a) maternal condition in: non-complicated pregnancy (NCP)- 151 cases, pregnancy-induced hypertension (PIH)- 37 and sustained chronic hypertension (SCH)- 25 and b) newborn weight in: small for gestational age (SGA)- 38 cases and adequate for gestational age (AGA)- 175. Of these placentas, 81 belong to the random sample, which was used to determine the frequency of VUE in the population studied. Eight blocks were taken from placental parenchyma and the histological sections were stained with hematoxylin and eosin. Frequency tables of categorical data were analyzed using the chi- square test and Fisher test; statistical significance was considered for p< 0.05. RESULTS: The frequency of VUE in the placentas was 30.8% in the random sample, 39% in NCP, 29.7% in PIH, 32% in SCH, 34.2% in SGA infants and 37.1% in AGA infants. Placentas of pregnancies with hypertensive disorders were more affected by villitis with parenchymatous component (PIH - 27.0%, SCH - 28.0%). This lesion was also the predominant villitis in the placentas of the SGA infants (31.5%). In contrast, in placentas of NCP and AGA infants, the principal kind of villitis was basal, not associated to a parenchymatous component (16.5% and 14.8% respectively). In the majority of the cases the villitis was mild. CONCLUSION: In the population studied, the frequency of VUE is high (around 30%). The lesion occurs in a similar frequency in placentas from NCP, PIH, SCH, SGA and AGA infants, but basal villitis not associated to a parenchymatous component affects mainly the placentas of NCP and AGA infants. It is possible that this kind of villitis could be an expression of an antigenic stimulation diverse from villitis with a parenchymatous component. PMID- 12724816 TI - [Nutritional condition in chronic renal failure patients treated by hemodialysis in Amazonas]. AB - BACKGROUND: Chronic renal failure patients frequently show malnourishment. OBJECTIVE: The food intake and the nutritional status of 165 chronic renal failure patients on maintenance hemodialysis were studied. METHODS: Nutritional status of 165 patients were studied by anthropometry, biochemical measurements, protein nitrogen appearance rate (PNA) and a average food intake. RESULTS: The results show that in mean the percent standard body weight, the body mass index (BMI), the mid arm muscle circumference (MAMC) and the triceps skinfold thickness (TSF) were adequate, however, 38% of women and 27% of men had the TSF bellow the 5th percentile; 39% of men and 2% of women presented MAMC bellow the 5th percent, showing that lose body fat in both sexs and the muscle stores in men. Energy intake was 29.5 +/- 10.2 Kcal/kg/day and 74% of the patients consumed less than 35 Kcal/kg/day, the recommended level for this population. Although the protein intake was in the recommended range (1.32 +/- 0.4 g/kg/day), 47% of the patients consumed less than 1.2 g/Kg/day. Mean serum albumin was 3.96 +/- 0.36 g/dL (normal value). According to TSF and of MAMC criteria (the reference was the 5th percentile), 74 (44.8%) patients were classified as malnourished. CONCLUSIONS: The frequency of malnutrition is high in our population and the poor food intake may contribute to this condition. PMID- 12724817 TI - [Cardiotocography in pregnancies with absent or reversed end-diastolic velocity in the umbilical arteries: analisys of perinatal outcome]. AB - OBJECTIVES: To study the correlation between cardiotocography parameters and perinatal outcome in pregnancies with absent or reversed end-diastolic velocity (AERDV) in the umbilical arteries. METHODS: One hundred and twenty-seven cases presenting with AREDV followed between 1993 and 2000 were selected for analysis. The last cardiotocographic tracing performed on the day of delivery was reviewed and the following parameters were considered: magnitude of long-term variability, presence or absence of acceleration, late deceleration, variable deceleration, prolonged deceleration, pseudo sinusoidal pattern and the classification in normal, suspected and abnormal tracing. RESULTS: A significant (p<0.05) association was found between abnormal tracing and acidemia at birth (pH inferior to 7.20) with 71.9%, first minute Apgar score inferior to seven (73.4%), newborn intubation at delivery (64.1%), and early neonatal death (20.3%). The absence of acceleration was associated (p<0.05) to acidemia at birth (58.8%), first minute Apgar score inferior to seven (67.7%), newborn intubation at delivery (51%) and neonatal death (29.4%). Low FHR variability (<5bpm) was associated to (p<0.05): acidemia at birth (88.5%), newborn intubation at delivery (69.2%), early neonatal death (34.6%) and neonatal death (42.3%). Late decelerations were significantly (p<0,05) related to acidemia at birth.(78.2%). The severe variable deceleration was associated to (p<0.05): acidemia at birth (79.3%), newborn intubation at delivery (69%) and early neonatal death (17.2%). Prolonged deceleration was associated to (p<0.05) newborn intubation at delivery (70.6%). Pseudo sinusoidal pattern was associated to (p<0.05) early neonatal death (60%). CONCLUSION: The AREDV represents a severe fetal compromise with high risk to neonatal morbidity and mortality, and correlation between cardiotocography abnormalities and adverse perinatal outcome was demonstrated. PMID- 12724818 TI - [Must the TNM staging of the renal cell carcinoma be modified again?]. AB - OBJECTIVE: The behavior of the renal cells carcinoma stage PT1 is not completely clarified. We studied the presence of factors after prognostics and tumoral size in the recurrence of survival of the sporadic kidney carcinoma after surgical treatment. METHODS: 120 patients followed after nephrectomy had been revised retrospectively 93 PT1, 9 PT2, 11 PT3, 7 PT4, It was analyzed survival and recurrence of the disease inside of three groups of tumors: Group 1: < 4cm, group 2: 4-7cm and group 3: > 7cm, and the prognostics factors above-mentioned evaluated were nuclear degree, microvascular invasion, presence of committed ganglia and sarcomatous degeneration. RESULTS: The frequency of adverse prognostics factors increase as the tumor size increase. In the group 1, we had only four tumors of high degree and only one shown microvascular invasion that does not committed ganglia or sarcomatous degeneration. In group 2 there was 16 tumors of high degree, 4 sarcomatoses, two with positive microvascular invasion and two with positive ganglia. In group 3, was found 18 tumors of high degree, 15 with microvascular invasion and 7 with positive ganglia and 5 sarcomatoses. There was statistical significance in the specific cancer survival (p=0.002) and free of illness (p=0.0002) between the three groups. CONCLUSION: The evolution of tumors PT1 is distinct for lesser tumors of 4 cm and 4-7 cm fitting the subdivision of these two groups in T1a and T1b. PMID- 12724819 TI - [Nutritional status and food intake assessment of climacterics women]. AB - BACKGROUND: To evaluate the nutritional status and food intake of climacterics women. METHODS: In a transversal study were evaluated 154 women matriculated in the Clinic of the Climacteric Woman of the Health Center of Public Health College of the S o Paulo University, Brazil. The analyzed variables were body mass index (BMI) and food intake (energy, percentual distribution of macronutrients, A and C vitamins, calcium and iron minerals). To analyse food caloric value and calcium and iron minerals, were utilized two age groups (35-50 y and 51-65 y). To analyse BMI, macronutrients percentage and A and C vitamins were utilized the total population (35-65 years of age). In all variables were calculated mean and standard deviation. RESULTS: The mean BMI was 29.3 kg/m2; (+/- 6.2 kg/m2;), with 75% of population above normality. About the energy, the results indicated adequacy to two age groups; about the distribution percentual of macronutrients, all were inadequate. In relation to the vitamins and minerals, all were inadequates, with exception of iron. CONCLUSION: The population studied presents risks to health owing to high value of BMI and inadequate intake of macronutrients, calcium, A and C vitamins. PMID- 12724820 TI - [Thalidomide: new perspectives for its use as antiinflammatory, immunossupressive and antiangiogenic drug]. AB - The new uses of thalidomide are reviewed. It has recently been used as antinflammatory, immunosuppressive, antiangiogenic, and antiviral agent. Although its mechanism of action is not yet understood, the advantage of its use in several diseases, such as erythema nodosum leprosum, multiple myeloma, and graft versus-host-disease is evident. Owing to its teratogenic properties, the use of thalidomide must be very well controlled. However, thalidomide has become a very important alternative, with new applications being studied. PMID- 12724821 TI - [Critical appraisal of sevelamer for the treatment of hyperphosphatemia in patients with chronic renal failure]. AB - OBJECTIVE: To evaluate in patients with chronic renal failure (CRF) the effectiveness and the costs of sevelamer, a cationic polymer calcium- and aluminum-free, that is a new gastrointestinal phosphate binder. METHODS: Literature review and critical appraisal of six clinical trials about the effectiveness and two economic studies of sevelamer in CRF patients. RESULTS: Sevelamer is an effective phosphate binder (used in a mean daily dose of 3.5 g three times per day with meals) and with similar effect as that obtained with calcium salts, without the adverse manifestations of the latter (elevation of calcium x phosphorus product, hypercalcemia, vascular and cardiac calcifications, etc.). Moreover, sevelamer reduced serum LDL cholesterol in around 30%. Despite the greater direct costs of sevelamer compared with calcium salts, the total costs may be lower due to the reduction of costs with clinical complications and hospitalizations. CONCLUSIONS: Sevelamer has important therapeutic value in CRF patients with hyperphosphatemia. Economic analyses should be performed in our setting to define the cost-effectiveness relationship of sevelamer. PMID- 12724822 TI - [Diagnosis and clinical management of urinary tract infection]. AB - A review about recent aspects on diagnosis and clinical management of urinary tract infection (UTI) is presented. There is a wide variation in clinical presentation of UTI which include different forms as cystitis, pyelonephritis, urethral syndrome and the clinical relevance of asymptomatic bacteriuria and low count bacteriuria that must be distinguished from contamination. Pathogenetic aspects concerning bacterial virulence as well as host factors in susceptibility to UTI as urinary tract obstruction, vesicoureteral reflux, indwelling bladder catheters, pregnancy, diabetes mellitus, sexual activity, contraceptive methods, prostatism, menopause, advanced age and renal transplantation are discussed. Diagnostic criteria and the most common tests utilized for differentiation between lower and upper UTI have been reviewed. The authors conclude that a careful evaluation of the underlying factors is required for the correct diagnosis of UTI and to prevent recurrence and that appropriate strategies and specific therapeutic regimens may maximize the benefit while reducing costs and adverse reactions. PMID- 12724825 TI - Itemized bill: novel method to audit the process of laparoscopic cholecystectomy. AB - One of the recurring obstacles to the successful completion of a medical audit cycle is the unavailability of accurate and complete information. This is particularly evident in the review of clinical processes, where the hand-written medical record is the source of information. We have attempted to bypass this information deficit by using information recorded primarily for financial transactions, using the itemized bill. The study was conducted in two parts. Initially information recorded as an itemized bill for the in-hospital process of laparoscopic cholecystectomies over a one-year period was analyzed. Areas for change in practice were identified, and recommendations were developed. These recommendations were presented to a multidisciplinary group consisting of consultants, residents, and nursing staff involved in caring for these patients. A clinical pathway was developed and implemented from these recommendations. One year after introduction, a review of the clinical pathway was undertaken using the same methodology. The in-hospital process consisted of 23 charge categories according to the itemized bill. Of these, 8 accounted for 95% of the total charge. The surgeon's fee and the anesthesiologist's fee accounted for 34% of the total; medical/surgical supplies, 20%; operating theater time, 17%; pharmacy, 7%; radiology, 5%; laboratory, 7%; and ward, 4%. Areas were identified in the latter 6 categories for change. Review of practice 1 year after implementation of the changes showed that a large number of recommendations were in place. We describe a new method for auditing the processes of medical care, using the itemized bill to adapt and use information primarily recorded for financial purposes. PMID- 12724826 TI - Risk and outcome of bowel resection in patients with incarcerated groin hernias: retrospective study. AB - The goal of this retrospective study was to evaluate factors that increase the risk of bowel necrosis and document the outcome of bowel resection in patients with strangulated hernias. We identified 102 patients (60 men, 42 women) who underwent surgical treatment for an incarcerated hernia at the Kartal Education and Research Hospital Emergency Unit between April 1997 and April 2001. Patients in group 1 required intestinal resection (n = 16), and patients in group 2 did not (n = 86). The median age of the patients was 53 years (range 3-96). Demographic and surgical data were obtained from the patients' charts and compared between the two groups. Women required bowel resections more often than men (p < 0.05). Patients older than 65 years and those with femoral or epigastric hernias required resection more often than patients younger than 65 years and those with inguinal, umbilical, or incisional hernias (p < 0.05 for all). Group 1 patients had a longer hospitalization and experienced more overall complications and wound infections than group 2 (p < 0.05 for all). In conclusion, incarcerated hernias are more common in men, but intestinal resection is required more often in women. The risk of intestinal resection is higher for patients with femoral hernias and those older than 65 years. Patients who undergo intestinal resection have a higher overall complication rate related to wound infections but not an increased risk of other complications or mortality. PMID- 12724824 TI - Current management of colon trauma. AB - This article offers a comprehensive review of colon trauma from World War I to the present. The process of evidence-based medicine was used to analyze the data from the past 25 years and define standards of care in the field. Where data are less compelling, recommendations and suggestions are provided for future research. Topics highlighted include destructive and nondestructive colon injuries, rectal injuries, on-table colonic lavage, colonic bypass tubes, risk factors, perioperative antibiotics, and colostomy closure. PMID- 12724827 TI - Chitin-silk fibroin interactions: relevance to calcium carbonate formation in invertebrates. AB - In mineralized tissues chitin is almost always associated with proteins, many of which are known to have chitin recognition consensus sequences. It has been observed in some mollusk shells that there is a well-defined spatial relation between the crystallographic axes of the crystals and the chitin fibrils. This implies that the chitin functions directly or indirectly as a template for nucleation of the mineral phase. It is thus of much interest to understand the exact nature of the interface between the chitin and the proteins at the molecular level in mineralized tissues. Chitin/silk fibroin interactions were studied in vitro at the molecular level using homogenous films composed of the two macromolecules. The results show that the silk fibroin intercalates between the molecular planes of the chitin, and that the interactions are mainly through the chitin acetyl groups. Published X-ray diffraction patterns and infrared spectra of mineralized tissue organic matrices, as well as infrared spectra reported here of the squid pen and lobster cuticle, all show that in vivo the chitin and protein are not intimately mixed, but exist as two phases. We deduce that there is an interfacial plane between them in which the interactions are through the amide groups. PMID- 12724828 TI - von Kossa staining alone is not sufficient to confirm that mineralization in vitro represents bone formation. AB - Numerous techniques are currently used to characterize biological mineralization in intact tissues and cell cultures; the von Kossa staining method, electron microscopic analysis (EM), X-ray diffraction, and Fourier transform infrared spectroscopy (FTIR) are among the most common. In this study, we utilized three of these methods to compare the mineralization of cultured fetal rat calvarial cells (FRC) and the osteoblast cell lines 2T3 and MC3T3-E1 with the in vivo mineral of rat calvarial bone. The cells were cultured with or without ascorbic acid (100 microg/ml) and beta-glycerophosphate (2.5, 5, or 10 mM betaGP), and harvested between 16 and 21 days (FRC cells and 2T3 cells) or at 30 days of culture (MC3T3-E1 cells). In the FRC cultures, maximal von Kossa staining was observed with 2.5 and 5 mM betaGP in the presence of 100 microg/ml ascorbate. FRC cells also showed some von Kossa staining when cultured with bGP alone. In contrast, maximal von Kossa staining for MC3T3-E1 cells was observed with 10 mM betaGP. Only the cultures of MC3T3-E1 cells that received both ascorbate and betaGP produced von Kossa positive structures. The 2T3 cultures produced von Kossa positive staining only upon treatment with ascorbic acid and betaGP, which was greatly accelerated by bone morphogenic protein-2 (BMP-2). FTIR was performed on the mineral and matrix generated in FRC, MC3T3, and 2T3 cultures, and the results were compared with spectra derived from 16-day-old rat calvaria. The mineral-to-matrix ratios calculated from FTIR spectra for rat calvaria ranged from 2.97 to 7.44. FRC cells made a bonelike, poorly crystalline apatite, and, with increasing betaGP, there was a statistically significant (P 5 log colony forming units per ml (CFU/ml) within 1 hour (h). Interesting trends were also observed when differences with respect to absorbance ratios when data was normalized. Results also showed that diacetyl (0.1%) caused more cellular leakage and death of S. aureus at 45 degrees C as compared to 37 degrees C, and that cellular leakage per se was not directly related to death. Diacetyl, however, caused rapid death of S. aureus and S. typhimurium in the presence of 30% sucrose and NZ-amine broth at 55 degrees C; > 7 log CFU/ml death within 1 and 2 h respectively. We also conducted a study with commercial starter distillate (7%) and proved its ability to control both organisms under the same test conditions within 1 h at 55 degrees C. It is concluded that diacetyl is a potential candidate for the control of foodborne pathogens especially under low water activity conditions such as those encountered during meat fermentation. PMID- 12724924 TI - HEp-2 cells exposed to glucocorticoids and LPS undergo organelle damage and apoptosis. AB - Glucocorticoids are potent anti-inflammatory and immunosuppressive agents that are known to affect T cell-mediated inflammation by the inhibition of cellular proliferation and cytokine production. The literature is lacking in explaining the mode of action of such agents on the epithelial cells. Therefore epithelial cells (HEp-2) were used to determine the effects of cortisol administration or cortisol in the presence of LPS on the cells metabolic functions. Cells were treated with physiological concentrations of cortisol or cortisol + LPS for periods of 24, 48 and 72 hours. After each phase cell number, cellular damage and cellular morphology were determined. The results indicated that cortisol and cortisol + LPS treated cells inhibited cellular proliferation as well as cellular MDA levels as early as 24 hours. Analysis of programmed cell death by apoptosis staining for Annexin V revealed that cortisol and cortisol + LPS treated cells had lower positive response. However, these differences do not take into consideration the reduction in cell number in the cortisol and cortisol + LPS treated cells. Overall, the results indicate that cortisol has a remarkable effect on HEp-2 cellular proliferation similar to the reduction seen in the literature for T-cells. In addition to reduction in cellular number the cell's ability to adjust to a bacterial challenge may be directly altered. This information is important for managing patients who are immuno-suppressed with s respiratory tract infections. PMID- 12724926 TI - Association of pesticide exposure and risk of breast cancer mortality in Mississippi. AB - Breast cancer is the most common form of cancer among women in the United States. Established risk factors include advancing age, early menarche, late menopause, positive first relative, late age of first birth and socioeconomic status. Mississippi has a combination of risk factors making it suitable for studying the pathways of breast cancer etiology. The purpose of this study was to analyze pesticide exposure and the risk of breast cancer mortality. Data for this study consisted of secondary analyses of the Mississippi age-adjusted breast cancer mortality aggregated by period (1970-1994). The total number of acres planted during 1997-2000 for each Statistical Economic Area (SEA) and by type of crop was used as proxy measure for pesticide exposure. Analyses by SEA revealed potential evidence for an association between pesticide exposure and risk of breast cancer mortality in three areas: Greenville, Corinth and Yazoo. The total number of acres planted was positively and significantly associated with female breast cancer mortality rate, and these associations differed by race and type of crop. The strongest correlation was between breast cancer mortality rate for white women and rice crops planted in Yazoo (rho = 0.674, p < 0.030). Moderate correlations were found between African-American breast cancer mortality rates and total acres planted in Corinth (rho = 0.667, p < 0.049), catfish crops in Greenville (rho = 0.648, p < 0.031), and although not statistically significant (p < 0.066) also with total planted acres in Greenville (rho = 0.573). In conclusion, there are moderate statistically significant associations between number of acres of crops planted and the mortality rate from breast cancer in Mississippi. The association varies by state economic area, race and type of crop planted. PMID- 12724925 TI - CYP1a1, HSP70, P53, and c-fos expression in human liver carcinoma cells (HepG2) exposed to pentachlorophenol. AB - Pentachlorophenol (PCP) is a widely used biocidal compound with several industrial, agricultural and domestic applications. Although it has been shown to induce systemic toxicity and carcinogenesis in several experimental studies, the literature is scarce regarding its toxic mechanisms of action. Recent investigations in our laboratory have shown that PCP induces cytotoxicity and transcriptionally activates stress genes in human liver carcinoma (HepG2) cells [1]. We hypothesized that PCP-induced expression of stress proteins may play a role in the molecular events leading to toxicity and tumorigenesis in liver cells. To test this hypothesis, we performed the MTT-assay for cell viability, and the Western Blot and densitometric analyses to assess the expression of cellular protein including CYP1A1, c-fos, HSP70, and p53. Data obtained from the MTT-assay indicated a strong dose-relationship with respect to PCP cytotoxicity. The LD50 was computed to be 23.0 +/- 5.6 micrograms/mL. Western Blot and densitometric analyses also demonstrated a linear dose-response relationship with regard to CYP1A1 expression within the dose range of 0-50 micrograms/mL. However, a biphasic response was obtained with regard to HSP70, c-fos, and p53 expression, showing a peak induction at 25 micrograms/mL, and a drastic reduction in protein expression at 50 micrograms/mL, probably due to cell death at higher level of PCP exposure. At lower level of exposure, PCP was found to be mitogenic. PMID- 12724927 TI - The synergistic effect of conventional and sustained delivery of antioxidants on LNCaP prostate cancer cell line. AB - Studies have been limited involving antioxidants as a mode of treatment for prostate cancer. The goal of this study was to develop alternative treatment and prevention for the invasive behavior of prostatic mutations by exploring the synergistic effects of several commercially available antioxidants. The specific aims were to examine the viability, proliferation, and morphology of human prostate cells in culture upon treatment of low and high doses of vitamin E, selenium, and lycopene alone or in combination. In phase I, groups were treated with physiological doses of the experimental agents. In phase II, the same groups were treated in combination to observe any synergistic effect. Upon treatment with selenium, lycopene, and vitamin E, vitamin E treated groups demonstrated the greatest effect on most assays in the study. Data obtained from this study did not demonstrate that the antioxidants would eradicate LNCaP prostatic carcinoma cells; however, the suppression of the metabolic pathways, cellular damage, and lowered prostatic specific antigen was observed. PMID- 12724929 TI - Morphometric analysis of MRC-5 fibroblast like cells exposed to intermittent UV radiation. AB - The goal of this study was to develop a tissue culture model with and without cellular protection (anti-oxidants) in an attempt to prevent or treat squamous damage as a result of free radical generation. The specific aim was to evaluate the morphological responses of MRC-5 fibroblast cells exposed to UV-radiation and vitamin E supplementation. The criteria for the morphological evaluation was as follows; cell and nucleus shapes, size, N/C ratio, nucleus pleomorphism, hydropic swelling, as well as other features or characteristics of membrane and cytoplasmic alterations. The experimental design was divided into two phases. In the first phase the cells were exposed to radiation for 30, 45 and 60-minute intervals, the morphological evaluation was assessed using Image Pro Digital Analysis technique. In the second phase, vitamin E was administered to the cells before and after exposure to 45 min of radiation. Data obtained demonstrated that, forty-five minutes of radiation exposure caused traumatic stress to the cells, swelling, cellular debris and fragmentation. However, treatment with vitamin E pre and post exposure provided protection for the cells. Vitamin E treatment resulted in less cellular aggregation, more abundant eosinophilic cytoplasm and an overall healthy appearance. The data suggest that the use of vitamin E may minimize skin damage by protecting the integrity of the fibroblast cells for up to 72 hours in culture. PMID- 12724928 TI - A comparative study of four serological tumor markers for the detection of breast cancer. AB - Breast cancer is currently the third most common cause of cancer in the world. Circulating tumor antigens are often used as a minimally invasive tool for noting breast cancer progression. The objective of this study was to compare four tumor antigens (CA 15-3, CA 27.29, alpha-fetoprotein [AFP], and carcinoembryonic antigen [CEA]) for their diagnostic efficacy in breast cancer patients. It was hypothesized that CA 15-3 would proved to be superior to CA 27.29, CEA, and AFP in assay performance. Tumor marker assays were performed according to the manufacturers' directions. Assays used in this study were CA 15-3 and CA 27.29 (Fujirebio Diagnostics/Centocor Inc.), AFP (Abbott Inc.), and CEA (Hybritech Inc.). A total of 554 patient samples were obtained from an area hospital, plus 200 healthy adult samples which were used for the determination of normal reference intervals. The patients included patients with no disease (184), with non-malignant disease (11), with breast cancer (87), and with other types of cancer (272). Diagnostic percent sensitivities for each marker were: CA 15-3 (63%), CA 27.29 (39%), CEA (22%), and AFP (22%). Diagnostic specificities for each marker were comparable, ranging from 80-88%. Analytical parameters were evaluated for the assays and compared favorably. We concluded that CA 15-3 was the best tumor antigen for use as a diagnostic aid and monitoring agent. PMID- 12724930 TI - A laboratory observation of some toxic effects of commercially available implant polyurethanes on HL-60 cancer cells. AB - We studied the interaction of HL-60 cells with films of commercial polyurethanes (PU). The cast films were dried in an oven at 60 degrees C for forty eight hours, washed in doubly distilled water, and soaked in fresh water for another 48 hours to ensure that all of the leachables including the dimethylacetamide used as casting solvent was removed before they were introduced to the cells. The HL-60 cells were cultured in the RPM1-1640, at 37 degrees C, 100% humidity and 5% CO2. Evaluation of the effects between the cells and the films, was done by counting the live cells using a hemacytometer and a light microscope. Preliminary analysis of our data have shown that while some brands of polyurethane can slow down or suppress the growth of the phagocytic type of cells, others can enhance the cell growth. It was concluded that this observation was new but should be investigated further to better understanding the interactions which may lead to the development of in vitro methods for screening the safety and stability of PU intended for implantation. PMID- 12724931 TI - The use of digital imaging technology to assess the pathogenesis of coronary atherosclerosis: the role of elastin. AB - Heart disease is directly associated with aging as well as progression of atherosclerosis. The vessels begin to stiffen with age. It is speculated that the increase in stiffness can occur as a result of either increase progression of atherosclerosis or possibly due to the deterioration of the elastic components of the arterial wall. Regardless of the mechanism, an increase in vessel stiffness can lead to significant increase in the pathophysiological progression of the disease. The overall objectives of this investigation were to evaluate the coronary artery obtained from cadavers in their 7th, 8th and 9th of life and characterized the level of atherosclerosis and to identify using special elastin staining techniques the involvement of fiber disruption in atherosclerosis. The coronary arteries were obtained from cadaveric donors at the University of Saskatoon (average age 81.7 years, range 77-92 years of age). The arteries were fixed, sectioned and stained for routine analysis as well as with an Elastin staining protocol. The arteries were screened and the level of atherosclerosis was measured as well as thickness changes within the arteries. Digital imaging was used to capture the areas of elastin disruption. The overall results suggest elastin disruption occurs as the atherosclerotic plaque progresses. The imaging system in conjunction with elastin staining allows for a very sensitive method to analyze the tissue for the progression of pathophysiological disease mechanisms. PMID- 12724932 TI - Morphometric evaluation of ovarian tissue exposed to PCB conventionally and in a sustained manner. AB - Most natural estrogens are short-lived, do not accumulate in tissue and are easily broken down in the liver. In contrast to natural estrogens, estrogenic drugs such as ethynylestradiol, diethylstilbestrol, synthetic environmental estrogens such as beta-hexachlorocyclohexane, polychlorinated biphenyls (PCBs), and phytoestrogens, are more stable and remain in the body longer than natural estrogens. Because most of these compounds are lipophilic, they tend to accumulate within the fat and tissue of animals and humans. Thus, depending on the natural estrogen levels, environmental estrogens may have different influences (mimicking, or blocking estrogen's effects) on estrogen activities. A total of 14 adult female rats were divided randomly into three groups. Animals in group I (n = 4) served as control. Animals in group II (n = 5) were injected with PCBs, and animals in group III (n = 5) were implanted with TCPL capsules containing PCBs. Pap smears were obtained daily, and at the end of the experimental phase the animals were sacrificed and vital organs as well as reproductive organs were obtained and wet weights were recorded. Significant reductions in ovarian wet weights were found in all animals treated with either sustained release of PCBs or injection of PCBs. Ovarian tissue was further analyzed histologically to determine the effects of PCBs. Histomorphometric data revealed significant reduction (p < 0.05) in the total ovarian area of animals treated with PCBs. Measurements of cross-sectional lengths confirmed the reduction seen in the area. Overall, the data suggest that PCB pollutants have produced detrimental effects on endocrine function as well as fertility regardless of the route of administration. PMID- 12724933 TI - Improvement of experimental allergic encephalomyelitis (EAE) by thymoquinone; an oxidative stress inhibitor. AB - Experimental Allergic Encephalomyelitis (EAE) is an autoimmune demyelinating disease of the central nervous system that is widely accepted as an animal model for the human multiple sclerosis. Oxidative stress appears to play a role in the onset and progression of EAE. We reasoned that decreasing oxidative stress might ameliorate symptoms and signs of EAE. Thymoquinone is reported to inhibit oxidative stress. One way of decreasing oxidative stress is to induce glutathione (GSH). We tested the impact of Thymoquinone (1 mg/kg, injected at tail vein) in our EAE model. We induced (EAE) in female Lewis rats using myelin basic protein emulsified with complete Freund's adjuvant. 24 animals were placed into three groups: A) Rats with EAE B) EAE rats with concomitant five day injection of Thymoquinone days 1-5, C) EAE rats with five doses of Thymoquinone injected at day 12-17. Twenty-eight days later, animals were sacrificed; spinal cord tissues collected for glutathione (GSH). RESULTS: 63% of animals in group "A" developed hind limb weakness and/or paralysis while 37% developed mild tail weakness, perivascular inflammation and low spinal cord GSH level. 25% of animals in group "B" exhibited mild tail and hind limb weakness and 75% animals had no symptoms, no perivascular inflammation and high spinal cord GSH level. 63% of animals of group "C" showed improving symptoms following Thymoquinone injections, no perivascular inflammation and higher GSH level while 37% of animals showed no symptoms prior and post Thymoquinone injections. Clinical symptoms correlated well with perivascular inflammation and GSH level. Animals received Thymoquinone at day 12-17 had higher GSH level, no perivascular inflammation and no symptoms compared with other groups. CONCLUSION: Thymoquinone inhibited oxidative stress which leads into improvement in our EAE animals. Thymoquinone may have a role in treatment of Multiple Sclerosis. PMID- 12724934 TI - Heterotopic bone formation: presentation of an experimental rat model and a clinical case. AB - Heterotopic bone formation (HBF), an ill-defined phenomenon, refers to the formation of bone in tissue that normally does not ossify. Two existing theories to explain HBF are that two cellular entities, one from the bone and the other from the muscle or fascia (two cell types) are involved, and that stem cell responds to a factor induced by trauma (one cell + factor). In our report, the HBF in a patient's vertical abdominal wound was possibly due to IEL, a stem cell, which is stimulated by a factor from xiphoid when it is traumatized by surgical incision. After 28 days' culture rat tissue specimens from the xiphoid, upper gastrointestinal tract, pubis and bladder exhibited macroscopic mineralization with cellular infiltration, a paradigm of 2-dimensional BF. Characteristically, pubis + bladder, xiphoid + ileum and xiphoid + duodenum showed 2-dimensional BF by as early as 5 days. Thus, it appears that both theories of HBF may be valid. PMID- 12724935 TI - Effects of static electromagnetic fields on characteristics of MG-63 osteoblasts grown in culture. AB - The effects of static electromagnetic fields (SEFs) on MG-63, a human osteoblast cell-line, were investigated. We examined proliferation, proline uptake and gene expression in an SEF approximately 1/728th the intensity of those previously reported. Cells were placed within an SEF apparatus (average field intensity of 0.618mT) with appropriate controls. Proliferation was measured by 3H-thymidine incorporation and showed a 34% decrease in cells exposed to SEF (P = .0001; N = 3). Proline, a major component of collagen necessary for bone formation by osteoblasts, incorporation was reduced 37% (P = 0.006; N = 3). Reverse transcription-polymerase chain reaction revealed that collagen I, alkaline phosphatase, parathyroid hormone-receptor, and osteocalcin mRNA's were down regulated with the low intensity SEF. Exposure to very low SEFs affects the MG-63 osteoblasts in a manner that may be detrimental to bone formation. PMID- 12724936 TI - Morphological evaluation of MRC-5 fibroblasts after stimulation with static magnetic field and pulsating electromagnetic field. AB - The quality of tissue repair and the speed with which that repair can be accomplished are the two major variables in the healing of any injury. Today, magnetic field exposure to traumatized areas has shown to be a promising tool in the healing process. The exact mode of action by which radiating and unchanging magnetic fields still has to be elucidated. The objective of this study was to evaluate the morphology of MRC-5 fibro-blasts after stimulation with static and pulsating magnetic fields. Under sterile environment, a total of 24 wells were loaded with 50,000 MRC-5 cells each and further divided into three groups. Groups 1 and 2 were exposed to magnetic fields, static and pulsating respectively. Group 3 wells were unexposed and served as the control group. The cells were monitored at 0, 24, 48, and 72 hours and representative views were captured using digital analysis techniques. The live cells were screened for cellular mobility, cell distribution, and cellular morphology (size, shape, lysis, and background). After 72 hours, the supernatants and cells of all three groups were collected and MDA analysis was performed to determine possible cellular damage. Group 1 cells continued to grow at a reasonable rate, but there was substantial cell membrane damage (high MDA levels, p < 0.05). Group 2 cells appeared to be very stressed under these conditions especially at the initial phase (24 hours). In conclusion, the use of pulsating magnetic stimulation can be beneficial in the healing process of soft tissues. PMID- 12724937 TI - Autoradiographic evaluation of electromagnetic field effects on serotonin (5HT1A) receptors in rat brain. AB - Serotonin (5HT1A) is a chemical mediator of inflammation and the largest single neurotransmitter system of the brain. Its secretion and physiological actions mediate stress and pain, affecting both immune and nervous system functions through the hypothalamic-pituitary-adrenal axis. Serotonin receptor dysfunction is well-characterized in mental disturbances like depression and anxiety. Transcranial magnetic stimulation has been used therapeutically to treat refractory disorders like non-responsive depression and may act in part through its effect on 5HT1A receptors. Previously we have shown that in vitro, 5HT1A receptor binding to a radioactive agonist can be modulated by specific intensity and frequency electromagnetic fields (EMFs). In the present report we have used quantitative receptor autoradiography to evaluate 5HT1A receptor density in rat brain and the impact of pulsed EMF exposure on receptor binding in key brain regions. Rats used in this study had whole body exposures to either a geofield control or to pulsed EMFs to evaluate the treatment for chemically-induced tendinitis. Since the brains were exposed coincidentally as a consequence of the main experiment, we investigated the potential for EMF-induced changes in areas such as the hippocampus. This pilot study should provide a detailed understanding of magnetic field effects on stress-responsive brain regions and will lead to a more coordinated approach to the use of such modalities for therapeutic intervention in humans. PMID- 12724938 TI - LDL oxidation by androgen exposure using MRC-5 cell line as a model. AB - The relationship between LDL oxidation and atherogenesis has not been fully investigated. The objectives of this study were to investigate the effect of androgens (testosterone) on the intactness of LDL upon incubation with MRC-5 cells and to evaluate the morphological changes associated with the incubation of androgens with MRC-5 cells. A total of 72 tubes plated with MRC-5 fibroblasts (50,000) were divided into four equal groups. Each of these groups were treated with one of the following: serum only (control), low-dose LDL, high-dose LDL, and LDL + testosterone. The experiment was divided into 24, 48, and 72-hour periods. At the end of the incubation periods, representative samples from each group were used for the determination of LDL oxidation. This was accomplished through lipoprotein electrophoresis. Results of the lipoprotein electrophoresis indicate that LDL was modified during the incubation periods with low-dose LDL, high-dose LDL, and LDL + testosterone. This was evident by the variations in electrophoretic mobility displayed by the treatment groups. Morphological evaluations were performed using light microscopy. MRC-5 cells treated with LDL + testosterone resulted in significant cellular damage. Results from this study suggest that MRC-5 cells are capable of modifying the native form of LDL. However, further investigation is needed to determine the exact mechanism of LDL modification and the role of MRC-5 cells in its modification. PMID- 12724939 TI - Cytopathological changes associated with continuous and intermittent exposure to glucocorticoids using raw cell line as a model. AB - In a world of pathogens, the ability to detect an incipient infection and to respond rapidly and appropriately to it, is a decisive part of our survival strategy. The innate immune system provides the first response to an infection by initiating an inflammatory response. The monocyte surface of the molecule CD14 is a key element in this response system. Over activation of the inflammatory response can lead to systemic inflammation and septic shock. Glucocorticoids are anti-inflammatory and immunosuppressive agents that act on many cells of the body, including monocytes/macrophages. The specific aim of this investigation was to compare the role of conventional versus sustained delivery of Cortisol on the viability and morphology of RAW 264.7 macrophages. RAW cells were obtained from American Type Culture Collection. Standard laboratory protocols were followed in cell plating (10(4) cells/well), phase terminating, and morphological evaluation. The experimental design consisted of two phases. Each phase had non-treated control groups and experimental groups. The results of this investigation yielded that: (i) TCP drug delivery system was capable of providing sustained and continuous release of Cortisol as evidenced by survival of cells in all experimental wells, and (ii) regardless of the Cortisol concentration, RAW cells were viable throughout the experimental periods. Overall, the results of this study provide a comparative analysis of the modes of actions of steriods; and, the results document the effects of the susceptibility of infection based on route. PMID- 12724940 TI - Histomorphometric evaluation of renal glomeruli exposed to sustained delivery of estrogen using adult ovariectomized rats. AB - Hormonal replacement therapy (HRT) has shown to be efficacious in treatment and preventing of heart disease, osteoporosis and reducing mortality in postmenopausal and ovariectomized females. Several attempts to utilize the native estrogen and its analogs such as Depo-Provera, conjugated estrogen and estrogen benzoate have shown different physiological responses. In addition, the route of administration and its mode of action is lacking in the literature. The specific objective of this study was to investigate the role of sustained delivery of estrogen on the functional and structural capacity of the kidney using adult female rats as a model. A total of 24 adult female rats were subdivided into four equal groups. Groups I and II were ovariectomized (OVX) by following standard laboratory surgical procedures. Each rat in groups II and III (intact) were implanted with tricalcium phosphate lysine (TCPL) drug delivery system loaded with 40 mg of estrogen. Rats in group IV were unimplanted and untreated to be served as a control group. At the end of 45 days post treatment the animals were sacrificed by using overdose of Halothane and assured by cervical dislocation. Vital and reproductive organs were retrieved, weighed and subjected to H&E staining procedure. The results of this investigation suggest: (i) TCPL delivery system released estrogen at a sustained level for 45 days without any untoward response, (ii) the wet weights of kidneys (normalized to body weight) were increased (p < 0.05) in intact rats treated with estrogen compared to control, (iii) sustained delivery of estrogen resulted in a maintenance of kidney weights compared to the control level, however, the lack of estrogen treatment resulted in a remarkable regression in the kidney weights of OVX rats, (iv) the ratio of renal arteries-diameter (normalized to arterial wall thickness) was significantly increased in intact rats treated with estrogen compared to the control and other experimental groups, (v) histopathological evolutions of renal tubules revealed tubular epithelial damage in intact rats received estrogen treatment compared to the control and other groups. In conclusion, this study suggests that exogenous estrogen therapy could lead profound tubular damage and consequently major renal insufficiency. HRT also could lead to hypertensive renal damage. PMID- 12724941 TI - Use of proteomics methodology to evaluate inflammatory protein expression in tendinitis. AB - In previous studies we established a rat model of acute tendinitis including functional and mechanical measures of healing. Achilles' tendinitis was induced by injection of collagenase, an enzyme that produces localized fiber digestion and edema formation. As quantitative measures of tissue inflammation, hypercellularity and edema were evaluated in injured tendons in comparison with controls. Using the rat tendinitis model, we have applied isotope-coded affinity tag analysis (ICAT) methodology to indicate localized tendon healing by quantitating protein expression. This novel proteomics method allows detection of subtle differences in protein levels that provide a detailed picture of tendinitis healing. The method involves a new class of chemical linkers used to differentially label cysteine residues from similar peptides in control and treated protein samples with heavy (deuterium off of backbone) and light (hydrogen off of backbone) ICAT reagents that are otherwise chemically identical. Proteins were extracted under liquid nitrogen from control untreated or injured Achilles' tendons 72 hours after collagenase-injection. These proteins were digested with endoproteinase Glu-C and trypsin and the resulting peptide mixtures were evaluated using reverse-phase C18 HPLC and Tristricine SDS-polyacrylamide gel electrophoresis. The two ICAT-modified peptide populations were mixed, affinity-purified and analyzed using microcapillary liquid chromatography and electrospray ionization tandem mass-spectroscopy. The process resulted in relative abundance and charge-to-mass ratio data used in conjunction with database searching to identify proteins expressed differentially in the two treatment groups. By analyzing different time periods in the healing process, an accurate model of the healing rat tendon can be made. PMID- 12724943 TI - Down regulation of CD14 expression through pretreatment with glucocorticoids. AB - Glucocorticoids such as cortisol are potent immunosuppressive agents that act on many cells of the body, including macrophages. Macrophages express CD14 in response to lipopolysaccharides (LPS) extracted from bacterial coats. The objectives of this study were: (1) to determine if pretreatment of macrophages with cortisol for 30 minutes prior to challenging the cells with endotoxin results in increased cell loss, cell damage (MDA), and suppression of CD14 receptors; and (2) to determine if CD14 receptor expression is able to recover with time. An experimental design incorporating RAW 264.7 cells (RAW) was used in order to evaluate our objectives. The cells were plated on 24 well plates and subsequently divided into four groups. The first group was untreated and served as the control. Group two was treated with LPS, group three with 10 uL of cortisol and a combination of LPS and cortisol was used in the treatment of the fourth group. The cells were recovered at 24, 48, and 72 hours. Results showed that there was a significant decrease in the proliferation rate in RAW cells exposed to cortisol and LPS either alone of in combination when compared to the untreated cells. Cell damage was also increased in treated cells. LPS caused receptor expression at all time points. CD14 expression was down regulated at 48 hours in cells pretreated with cortisol, however, this suppression was no longer evident at 72 hours. PMID- 12724942 TI - Cultured keratinocytes and dermal fibroblasts on a double-layer scaffold with bi medium culture system. AB - The increasing survival rate of massive burn patients, whose available donor sites for autografting are very limited, results in the emerging development and requirement of artificial skin [1-4]. The objective of this study is to produce cultured autologous keratinocyte grafts and to develop an optimal skin substitute for permanent wound closure. In the development of skin equivalent, human dermal fibroblasts were seeded to form three-dimensional dermal replacement tissue. The keratinocytes were initially cultured in keratinocyte serum free medium supplied with epidermal growth factor (EGF). After two days, the medium was changed to keratinocyte basal medium (without EGF) and subsequently cultured for 14 days by the air-liquid interface culture method. We found that time modulation of EGF has great effect on keratinocyte cell behavior. It is suggested that epidermal keratinocytes with bimedium culture system developed the basement membrane and also differentiated upward in the form of multi-layers. PMID- 12724944 TI - The analysis of the dorsal aorta and renal vessels exposed to sustained delivery of AED, T, and DHT using a rat model. AB - Studies have shown that endogenous estrogen minimized cellular injury at the organ level; however, very little research was done to determine the effects of endogenous androgens such as Testosterone (T), Dihydrotestosterone, (DHT), and Androstenedione (AED) on the cardiovascular system at the cellular level. Studies targeted at establishing such effects will broaden our understanding of the roles played by these male steroid hormones on the cardiovascular system. Our objective therefore was to (1) Use a Rat model and sustained delivery of physiological levels of these hormones to and (2) evaluate pathophysiological effects on the dorsal aorta and renal vessels. Sprague Dawley rats equally divided into four groups (n = 4) were included in the study. Group 1 animals served as control, groups II, III, and IV were treated with TCPL drug delivery devices containing 40 mg each of T, DHT, and AED, respectively. Animals were sacrificed after 90 days of exposure. Aorta and renal vessels were stained with hemotoxylin and eosin for histopathological evaluation. Results showed that TCPL drug delivery systems released 5 ng/ml/day of T, and 2 ng/ml/day of DHT and AED. After 90 days and evaluating each group, the renal arteries showed that groups exposed to T, DHT and AED showed an increase in the width of the measured arteries compared to the control group. The renal arteries exposed to AED showed the most significant increase in width. Compared to the dorsal aorta of the control group, T treated animals had decreased dorsal aorta width. The results revealed the following: (i) the exposure of sustained levels of androgenic hormones exhibited increased width of the renal arteries compared to the control animals; (ii) animals treated with sustained delivery of T had a decreased aortic width when compared to control animals; and (iii) anabolic receptors may be differentially expressed in the dorsal aorta and renal vessels of adult male rats. PMID- 12724945 TI - Primary rabbit epithelial cells exposed to adrenal, gonadal, and pituitary hormones in culture. AB - Various studies have investigated the effects of Monkey Kidney Epithelial cells to various agents but there are few studies that are looking at other species to determine the effects these agents may have. Studying other species and their effects may provide valuable information on the differences that are seen between humans and information as to why treatment works for some and not for others. The specific objective of this study was to investigate the effects of cortisol (C), testosterone (T), dehydroepiandrosterone (DHEA), estradiol (E), and Growth Hormone (G) individually and in combination on the proliferation and viability of Primary Rabbit Kidney epithelial cells in culture. In phase I: a total of 30 tubes plated with PRKEC were divided into six equal groups. In-groups 1-5, tubes were treated with physiological doses of hormones individually i.e., C, T, DHEA, E, and G respectively. Data obtained suggests that: (1) PRKEC treated with T for 24, 48, 72, and 96 hours displayed a significant increase in catalase, LDH, as well as MDA, (2) PRKEC treated with DHEA for 48 hours displayed a reduction in cell proliferation (70%) and an increase in MDA, LDH, and catalase. The hormones studied affect Primary Rabbit Kidney Epithelial cell growth, function, and viability, however the data suggests that further study needs to be conducted to determine the full impact of steroids and growth agents on Kidney Epithelial cells. PMID- 12724946 TI - Organotypic keratinocyte-fibroblast cocultures on a bilayer gelatin scaffold as a model of skin equivalent. AB - The skin consists of epidermis and dermis. In order to develop a biomimetic material for more realistic tissue regeneration, we fabricated a double layer gelatin sponge with different pore sizes by freeze-drying at different temperatures -20 [symbol: see text] and -196 [symbol: see text], respectively. A pseudo microporous structure, which provided the paracrine crosstalk in epithelial-mesenchymal interactions similar to the basement membrane in epidermal dermal junction was formed between this two compartment. Chrondroitin-6-Sulphate (C-6-S) and hyaluronic acid (HA) were incorporated within the gelatin scaffold to create an appropriate microenvironment for cell proliferation and migration. The mechanical strength of gelatin sponges was improved by cross-linking of gelatin with carbodiimide in 95% ethanol solution two times. The lattice structure and pore sizes were evaluated by SEM to confirm that the interconnected porous structures were retained. We chose a dynamic spinner flask seeding method for more even distribution followed by a culture system in the air-liquid interface cultured for 21 days. Differentiation and phenotypic expression of keratinocytes were investigated by histological analysis and immunohistochemical identification. In this study, we found multiple epidermis-like layers constructed by cultured keratinocytes. It is suggested that the bi-layer scaffolds have the potential to be used as skin equivalents for application in burn wounds. In the future, the qualitative of chemical reagents and in vivo animal model tests will be evaluated. PMID- 12724948 TI - Morphometric analysis of the hormonal effect on tissue-implant response associated with TCP bioceramic implants. AB - Recent studies in our laboratory have demonstrated the need for an experiment comparing the effect of steroid hormones on the cellular composition of the fibrous tissue capsules surrounding tricalcium phosphate (TCP) bioceramics. The effect of steroid hormones on inflammatory cells has been widely reported in the scientific literature. The purpose of this research is to contribute to the development of a long-term ceramic drug delivery system with human and veterinary applications. This study contributes information on the composition of fibrous tissue capsules surrounding hormone loaded TCP bioceramic implants at 90 days post-implantation. The data obtained in this study was gathered using protocols developed from ImagePro software. Sixteen animals in four experimental groups were implanted with one TCP bioceramic each. Group I animals were implanted with a sham TCP ceramic not containing a steroid hormone (control group). Group II animals received the testosterone loaded TCP ceramic. Group III animals were implanted with the dihydrotestosterone loaded ceramic. Group IV animals received the androstenedione ceramic. At 90 days post-implantation, the animals were euthanized. The implants and fibrous tissue capsules surrounding them were then extracted. The fibrous tissue capsules were evaluated microscopically using ImagePro software following routine II&E staining, modified Papanicolau, and Masson's trichrome. The research variables comparing the hormonal effects on fibrous tissue composition obtained by digital analysis were as follows: thickness of the fibrous tissue, quantification of macrophages, neutrophils, fibroblasts, vascularity, and area occupied by collagen. The results of this study support several conclusions. All three of the hormones in this study, particularly androstenedione, have significant affects on the thickness and cellular composition of the fibrous tissue capsules when compared to the control group. Digital analysis software enables more accurate and reproducible results to be obtained when specific procedures are followed. PMID- 12724947 TI - TCPL delivery system as a model for E + P replacement therapy in post ovariectomized adult rats mimicking a postmenopausal condition. AB - The specific aim of this investigation was to utilize TCPL delivery system as a model for estrogen (E) plus progesterone (P) replacement therapy in post ovariectomized adult rats mimicking a postmenopausal condition. A total of 20 adult female rats were used in this study. The animals were randomly divided into four different groups: groups II, III and IV were ovariectomized (OVX), OVX plus E (40 mg loaded TCPL), and OVX plus E (TCPL, 40 mg) plus P (TCPL, 80 mg) treatment. Group I animals (n = 5) served as intact control group. Blood samples were collected biweekly for 30 days. Vaginal smears were taken and screened daily during the entire investigation. The total serum levels of E, P, luteinizing hormone (LH), and follicle stimulating hormone (FSH) were measured by means of radioimmunoassay procedure. Data obtained from this study suggest the followings: (I) OVX resulted in an increase in total serum levels of LH and FSH within 2 days post-ovariectomy, (II) TCPL were capable of releasing sustained levels of E (10 40 pg/ml) and P (2.30-3.75 ng/ml) at the end of second day and continued until the 30th day, (III) the sustained levels of E plus P were able to suppress the post ovariectomy rise of LH and FSH to almost physiological levels, (IV) sustained delivery of E resulted in maturation of vaginal epithelium and the smears exhibited the estrus phase throughout the investigational period, and (V) sustained delivery of E + P induced no estrus and the epithelial changes resembled the OVX group. PMID- 12724949 TI - Immunohistochemical detection of cytokine expression in tissue-implant response associated with TCP bioceramic implants loaded with steroid hormones. AB - Recent studies in our laboratory have demonstrated the need for an experiment comparing cytokine expression in fibrous tissue capsules surrounding tricalcium phosphate (TCP) bioceramics loaded with androstenedione, dihydrotestosterone, and testosterone to that of a control to further elucidate the mechanisms involved in the tissue-implant response. The effect of steroid hormones on inflammatory cells has been widely reported in the scientific literature. The purpose of this research is to contribute to the development of a long-term ceramic drug delivery system with human and veterinary applications. This study contributes information on the presence of cytokines in fibrous tissue capsules surrounding hormone loaded TCP bioceramic implants at 90 days post-implantation. Sixteen animals in four experimental groups were implanted with one TCP bioceramic each. Group I animals were implanted with a sham TCP ceramic not containing a steroid hormone (control group). Group II animals received the testosterone loaded TCP ceramic. Group III animals were implanted with the dihydrotestosterone loaded ceramic. Group IV animals received the androstenedione ceramic. At 90 days post implantation, the animals were euthanized. The implants and fibrous tissue capsules surrounding them were then extracted. The fibrous tissue capsules were evaluated microscopically following routine H&E staining and IHC staining of antibodies to IL-1 beta, Il-2, IL-6, TNF infinity, and cell specific markers for CD-4 and CD14 positive cells. The results of this study indicate that these hormones, particularly androstenedione, limit the expression of cytokines and greatly affect the cellular composition of the tissue-implant response. PMID- 12724950 TI - Thyrotropin releasing hormone (TRH) potentiates the metabolic effect of norepinephrine (NE) in warm-acclimated lean and obese rats. AB - We have studied the effects of chronic peripheral infusion of thyrotropin releasing hormone (TRH) and norepinephrine (NE) alone and together. Do these hormones additively affect energy balance and body weight similarly in lean and obese animals? 20 each lean Wistar and obese (fa/fa) Zucker strain rats were studied. Food intake, body weight and resting metabolic rate (RMR) were measured over a 14 day period. We confirmed that in the obese animals, separately the hormones act as in the lean: NE stimulated RMR, but with parallel stimulation of appetite so that weight gain continued; TRH stimulated RMR but inhibited food intake so that rate of weight gain was reduced. TRH and NE together acted to further enhance RMR, but the inhibitory and stimulatory effects on food intake cancelled out. Thus a negative energy balance resulted in both groups; however, resultant weight loss was profound in lean but only moderate in obese. We suggest that the relative insensitivity of the obese to TRH and NE together reflects the inability of their brown adipose tissue to respond to thermogenic agents, even when chronically administered. PMID- 12724951 TI - Characterization of a real time H2O2 monitor for use in studies on H2O2 production by antibodies and cells. AB - It was recently shown that antibodies catalyze a reaction between water and ultraviolet light (UV) creating singlet oxygen and ultimately H2O2. Although the in vivo relevance of these antibody reactions is unclear, it is interesting that among a wide variety of non-antibody proteins tested, the T cell receptor is the only protein with similar capabilities. In clinical settings UV is believed to exert therapeutic effects by eliminating inflammatory epidermal T cells and we hypothesized that UV-triggered H2O2 production is involved in this process. To test the hypothesis we developed tools to study production of H2O2 by T cell receptors with the long-term goal of understanding, and improving, UV phototherapy. Here, we report the development of an inexpensive, real time H2O2 monitoring system having broad applicability. The detector is a Clark oxygen electrode (Pt, Ag/AgCl) modified to detect UV-driven H2O2 production. Modifications include painting the electrode black to minimize UV effects on the Ag/AgCl electrode and the use of hydrophilic, large pore Gelnots electrode membranes. Electrode current was converted to voltage and then amplified and recorded using a digital multimeter coupled to a PC. A reaction vessel with a quartz window was developed to maintain constant temperature while permitting UV irradiation of the samples. The sensitivity and specificity of the system and its use in cell-free and cell-based assays will be presented. In a cellfree system, production of H2O2 by CD3 antibodies was confirmed using our real time H2O2 monitoring method. Additionally we report the finding that splenocytes and Jurkat T cells also produce H2O2 when exposed to UV light. PMID- 12724954 TI - Ethical considerations in bioengineering research. AB - Biomedical science and engineering have made rapid advancements in the field of medicine over the past few decades. New ethical problems arising from this technology are influencing biomedical research more and more. It is disturbing that bioengineering professionals have had relatively little contact with moral and legal theory in light of these developments and particularly since they represent the forefront of new medical innovations. The objective of this communication is to introduce the study of bioethics and the use of principlism when examining bioengineering problems and dilemmas. Specific examples derived from actual proceedings, such as the Baltimore case, will alert scientists to the importance of misconduct in academic society. Cases will be used to illustrate how tools learned in this presentation are applied to analyze bioethical issues. New technology has a large social impact and is setting the standard of care for treatment. The health care system continually relies on researchers to produce improvements in patient therapy. Society will increasingly expect scientists to be morally responsible for the research they perform and uphold those virtues that ensure good ethical conduct. PMID- 12724952 TI - Calmodulin activity in aging rat heart. AB - Calcium has been known to play an important role in the aging process. Intracellular calcium is regulated by a number of calcium binding proteins including calmodulin. In cardiac tissue, calmodulin is known to regulate a variety of functions. Therefore, to identify possible relationships between calmodulin and the aging process, heart tissue was collected from rats aging 3 weeks to 1 year. Heart tissue was homogenized in a homogenizing buffer and processed for soluble and membrane fractions. These fractions were further subjected to heat treatment. The heat-treated extracts rich in heat stable proteins were used for different experiments. The calmodulin activity was determined as a measure of stimulation of activator deficient 3'-5' phosphodiesterase. The activity of calmodulin at different ages was compared, and no significant changes were observed between 3 weeks vs. 8 weeks, 3 weeks vs. 6 months, and 3 weeks vs. 1 year. The soluble fractions also followed a similar trend, but there was a significant difference between 3 weeks vs. 1 year. At this point we do not know whether the decrease in activity is due to lower expression of calmodulin or decreased ability to stimulate the target enzyme. However, these data suggest that calmodulin may have some association with the aging process. PMID- 12724953 TI - Human embryonic stem cell research: an ethical controversy in the US & Germany. AB - Stem cells refer to a broad class of cells with potential for prolonged self renewal and the potential to differentiate. Human embryonic stem (ES) cells are derived from a fertilized human egg, before it has been implanted into a womb. Recently published research has used human embryos, produced by in-vitro fertilization for clinical purposes, to develop lines of pluripotent human ES cells. This review discusses the legal status and ethical implications of ES cell research with special attention paid to the situation in the USA and Germany. PMID- 12724955 TI - Validity of self-assessment outcome questionnaires: patient-physician discrepancy in outcome interpretation. AB - Patient outcome following total hip arthroplasty (THA) was evaluated using a previously described patient assessment outcome index questionnaires. The questionnaire was distributed to 263 patients who underwent cementless THA. One hundred and three patients responded to the self-administered questionnaire and had updated clinical evaluation. We obtained a modified Harris Hip Score (HHS) based on patient assessments of their own pain and function and compared it with the clinical HHS obtained at the patients' last office visit. The mean follow up period was 4 years. Statistical analysis was performed between the two scores. The correlation between the scores from the self-administered questionnaire and the patients' last office visit revealed a fairly low correlation coefficient (r = 0.467, p < 0.001). Relative lack of correlation between the HHS's obtained from these two sources was especially noted for patients with a pain score of 30 points or less. These 26 patients were subsequently interviewed in detail about their pain to further explain these differences. The etiology of the perceived "hip pain" was found to be secondary to trochanteric bursitis in 13 patients, lumbar spondylosis in 7 patients, arthrosis of the contralateral hip in 5 patients, and from other causes in 8 patients. Pain that was hip related (anterior thigh or groin) was present in only 5 out of the 26 patients with a pain score of 30 or less. Another source of discrepancy between the total scores of the HHS was found to be on behalf of the physician in evaluating the presence of a limp. We also found that patients' expectations had changed from their preoperative expectations. Although outcome measures developed and administered by clinicians are subject to bias from several sources, results of this study suggest that self administered patient outcome measures also have their limitations. The validity of self-administered patient outcome questionnaires can be severely impacted by the patients' understanding of the questions asked, as even the most seemingly simple questions are subject to misinterpretation. PMID- 12724956 TI - Life's a switch. Experiences in NSF undergraduate design projects. AB - During the summer of 2002 Stephanie Popp and Jennifer Barnes developed a manual, "Life's a Switch," through a project funded by the National Science Foundation. This manual teaches people how to build their own cost effective assistive switches. Assistive switches are a form of assistive technology which includes any device that enhances a person's quality of life by improving the individual's mobility, ability to perform daily activities, enhancing communication, or allowing participation in education, vocational activities and recreation. One main goal of assistive technology is to provide opportunities for children with disabilities to explore, play, learn, and communicate with others. Switches are essential tools used to provide these opportunities. When a child with developmental disabilities understands the connection between the activation of a switch and the resulting action it triggers, the knowledge of cause and effect is gained. Therefore, the basis for all future learning is established [1]. One of the current problems facing assistive switch users is the cost of available items. This project provides more affordable solutions for switch users by teaching the families and educators of switch users how to make their own switches and adaptors in the "Life's a Switch" manual. For example, some assistive technology vendors sell large button switches from $25.00 to $45.00, tread switches for $40.00, and pillow switches for $35.00 [2]. Amazingly, all parts and tools used to make these assistive switches can be bought and made into personally designed assistive devices averaging a cost of around $10.00 [3]. A workshop to teach this manual was also developed. This workshop will spread awareness of the more affordable options this project sets forth. In September of 2002, the first workshop was held in a laboratory classroom at the University of Wyoming's College of Engineering. Each attendant was provided with a kit that included all essential tools and components needed to make an assistive switch. Workshops scheduled into 2003 will provide educational opportunities for participants as well as opportunities for improvement of the manual. PMID- 12724957 TI - University of Wyoming, College of Engineering, undergraduate design project: star tracer. AB - The University of Wyoming received funding in the spring of 2002 from the National Science Foundation Division of Bioengineering and Environmental Systems in order to complete undergraduate design projects. One design project that was chosen by the College of Engineering involved partnering with the College of Education. The College of Education's Special Education Department needed some visual teaching aids to be redesigned and then built. Two undergraduate students were hired throughout the summer of 2002 under NSF REU funding in order to develop thirty new teaching devices. These devices were going to be used to educate middle school students about the effects of possessing a learning disability. The teaching aids are specifically designed for simulating the affects of dyslexia. The new teaching aids required improved transportability and durability, quicker setup time, and a lighter weight. Throughout the summer, the teaching aids were redesigned and built by an undergraduate student team from the College of Engineering, and have since provided many benefits for the state of Wyoming. PMID- 12724958 TI - University of Wyoming, College of Engineering, undergraduate design projects to aid Wyoming persons with disabilities. AB - In Spring 2002 the University of Wyoming received NSF funding from the Division of Bioengineering and Environmental Systems to provide a meaningful design experience for University of Wyoming, College of Engineering students that will directly aid individuals with disabilities within the state of Wyoming. Other universities have participated in this very worthwhile program [1, 2, 3]. To achieve the program purpose, the following objectives were established: Provide engineering students multi-disciplinary, meaningful, community service design projects, Provide persons with disabilities assistive devices to empower them to achieve the maximum individual growth and development and afford them the opportunity to participate in all aspects of life as they choose, Provide engineering students education and awareness on the special needs and challenges of persons with disabilities, and Provide undergraduate engineering students exposure to the biomedical field of engineering. To accomplish these objectives the College of Engineering partnered with three organizations that provide education and service related to disability. Specifically, the college has joined with the Wyoming Institute for Disabilities (WIND) assistive technology program, Wyoming New Options in Technology (WYNOT) and their Sports and Outdoor Assistive Recreation (SOAR) project along with the university's Special Education program. In this paper we will describe how the program was created, developed, and its current status. PMID- 12724959 TI - Advanced clinical engineering workshops in Central America. AB - Over the past ten years the American College of Clinical Engineering has conducted numerous Advanced Clinical Engineering Workshops in developing countries around the world. Most of the recent workshops have been conducted in Latin America and the Caribbean in cooperation with the Pan American Health Organization, an affiliate of the World Health Organization. This presentation describes workshop objectives and topics. It also describes the author's participation in workshops conducted in Costa Rica (2002) and El Salvador (2003). PMID- 12724960 TI - Self-reflecting: reclaiming our nursing roots. PMID- 12724961 TI - Unique outcomes of women and men who were abused. AB - PROBLEM: To determine if individuals who have experienced extensive victimization throughout their lives tell stories about "unique outcomes." METHODS: An examination of existing narrative data collected from 27 women and 17 men who had participated in one of several qualitative studies of sexual violence. Unique outcomes stories identified from the interview transcripts were categorized according to the type of experiences described, and the nature of men an women's stores were compared. FINDINGS: Six types of unique outcomes stories were identified in the women's narratives (rebellion, breaking free, resurgence, refuge, determination, confidant) and three types in the men's narratives (reawakening, buddy and normal guy, champion). CONCLUSIONS: Unique outcomes stories are common in narratives otherwise focused on abuse. Common themes are apparent, and the nature of men's and women's stories differ markedly. PMID- 12724962 TI - Aftermath of a patient's suicide: a case study. AB - TOPIC: Nurse psychotherapists often feel poorly prepared to cope with a patient's death by suicide. The psychotherapist may identify with the family, feel sad at the death, and be plagued by feelings of guilt and responsibility. PURPOSE: A case study illustrates the meaning of the loss to the therapist and the influence on professional identity, self-confidence, and self-esteem. SOURCES: Case study and review of the literature from Medline, psychinfo, and CINAHL. CONCLUSIONS: Therapists experience their own grief as a lack of omnipotence over suicide, and the fear of their colleagues' responses. Understanding bereavement and factors influencing bereavement may help therapists facilitate and reduce negative consequences of their own grief. PMID- 12724963 TI - Health outcomes for people with serious mental illness: a case study. AB - PURPOSE: To describe outcomes of clients with serious mental illnesses who used regional public mental health services; and what the public mental health system did well or needed to change to better satisfy clients' needs. METHODS: A modal case study analysis method was used with 15 clients. Participants were interviewed three times. FINDINGS: Participants reported dissatisfaction with their social functioning and general health, and unmet needs for case management services, social/recreational activities, and vocational rehabilitation. CONCLUSIONS: The case study method is an effective way for nurses to evaluate public mental health systems and to determine areas for system intervention. PMID- 12724965 TI - Aromatherapy and the use of scents in psychotherapy. PMID- 12724964 TI - Questions and answers for the advanced practice psychiatric nurse. AB - Because of the importance of maintaining current knowledge of technological advances involving genetics, neuroscience, and their impact on psychopharmacology, we have created a new column for advanced practice psychiatric nurses. This new column offers a question-and-answer forum that can help nurses maintain their knowledge of advances in prescribing and psychopharmacology, and implications for safe psychiatric care. Deborah Antai Otong has a wealth of knowledge and expertise with prescriptive authority and as a psychotherapist. She currently manages the care of patients with various psychiatric disorders including mood disorders, schizophrenia, dual diagnosis, and anxiety disorders. She is the author of numerous refereed journal articles and book chapters that focus on psychopharmacology and is a guest lecturer at a local university on this topic. She is also the author of several books, including Psychiatric Nursing: Biological and Behavioral Concepts (Clifton Park, NY: Delmar & Thompson Learning, 2003) and Psychiatric Emergencies (Eau Claire, WI: PESI, 2001). PMID- 12724969 TI - "The Campaign for Nursing's Future". PMID- 12724966 TI - Panic disorder: a tightening vortex of misery. PMID- 12724971 TI - [Paramedical personnel and diabetic care networks give hope]. PMID- 12724972 TI - [Which quality criteria for diabetic care networks?]. PMID- 12724973 TI - [Improving practices in the heart of care networks]. PMID- 12724974 TI - [Coordination nurse, a new job?]. PMID- 12724975 TI - [Screening program for foot lesion risk in diabetics]. PMID- 12724976 TI - [Role of the podiatrist in a care network]. PMID- 12724977 TI - [Diabetic foot ulcer]. PMID- 12724978 TI - [Dietetics and primary care]. PMID- 12724979 TI - [Group therapeutic education in a rural milieu]. PMID- 12724980 TI - [Which financing for quality therapeutic education? Interview by Bernadette Fabregas]. PMID- 12724982 TI - [Nursing shortage, a race against time]. PMID- 12724983 TI - [Fifteenth congress of the Society for Research and Education of Nurses]. PMID- 12724984 TI - [Follow-up of diabetic patients, a good investment]. PMID- 12724985 TI - [Depression, migraine, joint diseases: primary prevention]. PMID- 12724987 TI - [Decubitus ulcers in oncology, first results of a study]. PMID- 12724986 TI - [Utility of nursing consultations after hospitalization for patients on insulin]. PMID- 12724988 TI - [Leg ulcers, new therapeutic advances evaluated]. PMID- 12724989 TI - [Myocardial infarction]. PMID- 12724990 TI - ["Reinforcing primary prevention and therapeutic education" Interview by Bernadette Fabregas]. PMID- 12724991 TI - [Nursing care of non-traumatic thoracic pain]. PMID- 12724993 TI - [Management of myocardial infarctions in the cardiology interventional unit]. PMID- 12724995 TI - [The heart of symbolism, the symbolism of the heart]. PMID- 12724994 TI - [Management in cardiology intensive care units]. PMID- 12724996 TI - [Psychological management of the coronary patient]. PMID- 12724997 TI - [Life after an infarction: the dietician's role]. PMID- 12724998 TI - [Physical activity program for life]. PMID- 12724999 TI - [Heart 2001, a occupational preventive action in coronary risk patients]. PMID- 12725000 TI - [Best practice drug therapy. B. Use of antibiotics 8/urinary infections]. PMID- 12725001 TI - [2/10 alcohol]. PMID- 12725002 TI - New Jersey nursing--the first one hundred years. PMID- 12725004 TI - Complying with HIPAA regulations: a brief primer for nurses. PMID- 12725008 TI - Posttraumatic stress responses in children with life-threatening illnesses. AB - Posttraumatic stress symptoms have been reported in response to various serious medical illnesses in adults and children. Not surprisingly, posttraumatic stress is probably more common in response to acute, life-threatening, events that are related to the illness. Emerging data suggest that children often experience life saving medical procedures as traumatic, which puts caretakers and medical personnel in the role of perpetrators for the children. Trauma symptoms are also reported as common and severe in caregivers. Both of these issues have been previously poorly understood and should be addressed in assessment and treatment. As with other traumatic events, developmental considerations, the nature and severity of the event itself, social supports, and premorbid exposure to negative life events are also important issues to consider in developing appropriate interventions. The importance of developing prevention and treatment for PTSD in medically ill children and adults includes increased morbidity and mortality (e.g., nonadherence to medications) and psychiatric sequelae and decreased quality of life. Obstacles to systematic study of a psychiatric intervention for this group include difficulties assessing multidrug regimens and cognitive treatment effects in this group. The relative stability of social supports and the potential use of preventive measures make this an attractive population for intervention. Clinicians and researchers are encouraged to work together to develop and use uniform screening and assessment methods that will help to identify cases and facilitate the multicenter trials that are vital to increasing knowledge in this patient population. PMID- 12725007 TI - Posttraumatic stress disorder in children: diagnosis, assessment, and associated features. AB - Although a growing body of research has increased knowledge of the after-effects of trauma in children, including the development of PTSD, there are significant gaps in this knowledge concerning diagnosis, assessment, and associated features. With regard to diagnosis, variations in symptomatic expression of PTSD in children have been proposed; however, there have been few examinations of the validity of these variations in terms of course and consequence of PTSD defined in these ways. Several factors increase children's risk for development of PTSD or PTSD symptoms after trauma. Such information is potentially useful for identifying children who may benefit from more thorough or ongoing assessment after trauma. With regard to assessment, an array of increasingly sophisticated and clinically useful measures is being developed; however, currently there is a dearth of high quality data concerning the diagnostic use of different assessments. An area of general weakness concerning these assessments is the limited data on discriminant validity. A sizeable body of evidence indicates that trauma can produce diverse reactions in children, including a general increase in internalizing and externalizing symptoms. Clinically useful measures allow PTSD to be differentiated from this general reaction to traumatic events, much of which may reflect a nonpathologic response. With regard to associated features, limited information suggests that PTSD can have a cascading negative effect on children's development and functioning. More research is needed, however, to delineate factors that reflect risk for PTSD after trauma, factors that reflect consequences of PTSD, and mediating variables. PMID- 12725009 TI - Maltreatment and trauma: tracking the connections in adolescence. AB - Posttraumatic stress disorder is one of the most common and often prolonged consequences of childhood maltreatment. In this article the authors consider theories of trauma continuity, with emphasis on a relational path to maladjustment that links childhood maltreatment to elevated trauma symptomatology and intimate victimization in adolescent dating relationships. PMID- 12725010 TI - Dissociative symptoms in posttraumatic stress disorder: diagnosis and treatment. AB - This article explores the complex relationship between dissociation and psychiatric trauma. Dissociation is described as a defense reaction, a risk factor for the development of posttraumatic stress disorder, and as a set of syndromal disturbances. The authors discuss various models proposed for the relationship between these. They outline developmental considerations in diagnosis and treatment and end by discussing further needed research. PMID- 12725011 TI - Pharmacologic treatment approaches for children and adolescents with posttraumatic stress disorder. AB - Posttraumatic stress disorder is a common cause of morbidity in children and adolescents. The disorder in youth is similar to that in adults, with high rates of psychiatric comorbidity. Children seem to be more sensitive to the effects of trauma, and early life trauma exposure may induce a complex sequence of events that leads to the development of multiple psychiatric disorders in adulthood. The state of knowledge regarding medication treatments for children and adolescents is in the earliest stages of development. There are no well-conducted, randomized clinical trials to guide practitioners. Medication may play an important role in reducing debilitating symptoms of PTSD and providing a buffer for children while they confront difficult material in therapy and may help to improve their general functioning in day-to-day life. Given the various medications with potential usefulness in PTSD, it is helpful to use a stepwise approach to treatment. As a general principal, broad-spectrum agents, such as the SSRIs, are a good first choice. The SSRIs have efficacy in treating the core symptoms of PTSD and conditions such as the anxiety disorders and depression that commonly co-occur with PTSD. These agents also improve social and occupational functioning and an individual's perception of improved quality of life [41, 45, 46]. Although the SSRIs are generally effective for a broad spectrum of problems, clinicians should systematically monitor for the persistence of symptoms that do not respond to these agents. For example, despite significant improvements in core PTSD symptoms in one study that used sertraline, little improvement was seen in patients' comorbid anxiety and depressive symptoms [41]. This finding demonstrates the value of continuous symptom monitoring and shows that residual or comorbid symptoms may require a different medication to augment effective SSRI treatment for PTSD. A reasonable approach is to begin with a broad-spectrum agent, such as an SSRI, which should target anxiety, mood, and reexperiencing symptoms. Adrenergic agents, such as clonidine, used either alone or in combination with an SSRI may be useful when symptoms of hyperarousal and impulsivity are problematic. Supplementing with a mood stabilizer may be necessary in severe affective dyscontrol. Similarly, introduction of an atypical neuroleptic agent may be necessary in cases of severe self-injurious behavior, dissociation, psychosis, or aggression. Comorbid conditions such as ADHD should be targeted with pharmacotherapy known to be effective, such as psychostimulants or newer agents such as atomoxetine. Pharmacologic treatment of PTSD in childhood is one approach to alleviating the acute and chronic symptoms of the disorder. Despite the lack of well-designed, randomized, controlled trials that support efficacy, medication can be used in a rational and safe manner. Reduction in even one disabling symptom, such as insomnia or hyperarousal, may have a positive ripple effect on a child's overall functioning. Pharmacotherapy is typically used as one component of a more comprehensive multiple modality treatment package, including psychoeducation of the parent and child, focused exposure-based psychotherapy with adjunctive family therapy when indicated, and long-term booster interventions that use an admixture of psychodynamic, cognitive-behavioral, and pharmacologic interventions. PMID- 12725013 TI - The neurobiology of childhood trauma and abuse. AB - During the past decade there has been rapid progress in the understanding of the effects of exposure to traumatic life experiences on subsequent psychopathology in children. Trauma exposure affects what children anticipate and focus on and how they organize the way they appraise and process information. Trauma-induced alterations in threat perception are expressed in how they think, feel, behave, and regulate their biologic systems. The task of therapy is to help these children develop a sense of physical mastery and awareness of who they are and what has happened to them to learn to observe what is happening in present time and physically respond to current demands instead of recreating the traumatic past behaviorally, emotionally, and biologically. PMID- 12725012 TI - Long-term effects of childhood abuse on brain and neurobiology. AB - Early stress is associated with long-term alterations in brain circuits and systems that mediate the stress response. Early stressors have lasting effects on the HPA axis and norepinephrine systems. Other brain systems that are involved include benzodiazepine, opiate, dopaminergic, and various neuropeptide systems. These neurochemical systems modulate function in brain regions, including the hippocampus, amygdala, and prefrontal cortex. Long-term alterations in these brain regions are hypothesized to play a role in the maintenance of PTSD, depression, and other psychiatric symptoms after childhood abuse. PMID- 12725014 TI - Developing a culturally and ecologically sound intervention program for youth exposed to war and terrorism. AB - This article describes the public mental health approach used to develop and implement a school-based postwar trauma/grief intervention program for adolescents in Bosnia-Herzegovina. This approach includes development of multilateral partnerships with local and ministerial stakeholders, systematic assessment that yields a detailed understanding of the specific range and severity of trauma and loss experiences, current adversities and trauma reminders among the affected population, and a training program aimed at developing the capacities of local service providers and an indigenous support infrastructure so that the intervention program may be directed and sustained by people within the communities served. Concluding comments detail an expanded conceptual framework for public mental health interventions that may be appropriate for terrorist and mass-casualty events. PMID- 12725015 TI - Facing war, terrorism, and disaster: toward a child-oriented comprehensive emergency care system. AB - The combination of the overwhelming nature of disasters and the massive losses they engender gives rise to a complex clinical and social picture with longterm physical, psychological, and social effects on children, families, and communities. The authors suggest that to assess the damage properly, implement interventions on a large scale, keep tabs on rising needs, and restore societal function, mental health professionals must adopt an ecologic systems approach. This approach entails working within and together with related institutions (education, health, local government) and assisting other committed professionals within these institutions to mediate care. This is of utmost importance in the area of children's care because of their particular vulnerability and their special importance for families and society. For this reason, the authors suggest that emergency mental health systems be better designed and implemented while keeping children at the center of their focus. An essential component of the ecologic systems approach is improved education for mental health professionals, providing them the appropriate tools to cope with widespread disaster and the expertise to apply these tools. This approach, however, is not enough. A good outcome cannot be achieved without preparedness on the part of the other relevant institutions and the community as a whole. Greater awareness is needed among local and national authorities of the importance of metaadaptive systems and of local, national, and international networking. In the current global village that is threatened by pervasive terrorism, no community must face it alone. The challenge of a disaster to one community is a challenge to all. By working together we can lessen the devastating impact of these events, save countless lives, prevent untold suffering, and maintain hope for a better world for children. PMID- 12725016 TI - School reactivation programs after disaster: could teachers serve as clinical mediators? AB - Mental health interventions are known to prevent the progressive worsening of symptoms in young victims of disaster and, subsequently, to prevent a decline in their academic performance and self-esteem [8,46]. The tremendous needs that emerge after a disaster and the reluctance shown by most victims to seek professional help require mental health leaders to adopt a proactive stance and implement relief programs in the child's most natural setting. The school as institution and the teachers as empowered mediators offer the appropriate conditions for implementing an effective large-scale intervention program. Well intentioned child professionals who deal with school administrators and teachers must take into account that, as stated by Pfefferbaum et al [25], "avoidance is at the core of the posttraumatic response, and it sometimes involves avoidance of treatment." For child mental health professionals, routine collaboration across systemic boundaries may prove critical for the rapid mobilization of resources during mass traumatic emergencies. Further studies are needed to identify the protective and risk factors that predict resilience and pathology, respectively, and factors that facilitate or aggravate factors that predict improvement, resistance, and deterioration in response to treatment. PMID- 12725017 TI - [Current concepts of pharmacological treatment in schizophrenia]. AB - The modern concepts of pharmacotherapy of schizophrenia, with the special emphasis put on new antipsychotic drugs were presented. The attention was put on the need for careful evaluation of the patient's state: the diagnosis, treatment which the patient receives and compliance with the treatment. It was stated that modern concepts of treatment of schizophrenia do not solely rely on the use of modern drugs, but include the multidirectional therapeutic actions with an active participation of the patient and his/her relatives. PMID- 12725018 TI - [The relevance of genetically determined polymorphism CYP2D6 in psychopharmacology; the relationship between genotype and phenotype]. AB - Based on literature review the paper presents some clinical aspects of the genetically determined polymorphism of the CYP2D6. One of the main biotransformation processes of psychotropic drugs is oxidation catalysed by enzymes of cytochrome P-450. CYP2D6 is an isoenzyme of cytochrome P-450. Its activity is determined genetically and is characterised by interindividual variability. Genetically determined polymorphism of CYP2D6 is related to mutated alleles that code enzymatic proteins with different activity. Based on individual ability to oxidize drugs by CYP2D6 in population there are four phenotypically different groups: extensive (EM), ultra-rapid (UM), intermediate (IM) and poor metabolizers (PM). Each phenotype is determined by a given genotype. About 6-10% of the Caucasian population is known as PM phenotype. Drugs used in standard doses in this group may reach a markedly higher level in blood, even a toxic level. Compared to the group with EM phenotype persons with PM or IM phenotype are more likely to suffer from side effects that are related to impaired metabolic pathways that are catalyzed by CYP2D6. In the group with UM phenotype (1-7% of population) metabolism is very rapid, thus they need higher doses of psychotropic drugs to reach therapeutic blood level of drug. PMID- 12725019 TI - [Perspectives in the development of the concept of digesting mental information in psychotherapy]. AB - Modified Kepinski's concept of information metabolism constitutes a basis for the integrating model of psychotherapeutic diagnosis. It includes estimation of: boundaries, self-control, assimilation of new information ability, accommodation of existing schemata to new information, ability for elimination of unnecessary information, energetic resources. Outlines of concepts of psychopathology and psychotherapy there are in his publications. They have potential of general theory of psychotherapy which may constitute a background of integration of variety contemporary psychotherapeutic approaches. PMID- 12725020 TI - [On controversies around the Oedipus Complex]. AB - In the article the author would like to draw the readers' attention to one of the central issues of psychoanalysis, i.e. Oedipus complex. In the development of the psychoanalysis the classical conception of Oedipus complex presented by Freud gave rise to a severe criticism both on the part of some psychoanalysts, especially women such as e.g. Melanie Klein, and on the part of ethnographers such as B. Malinowski. In the paper the author presents the main controversies concerning this interesting discussion. The fact of the matter is that the Oedipus complex is a hotly debated subject, not only from the cultural point of view but also within the psychoanalytical movement. PMID- 12725021 TI - [Child abuse, neglect and pregnancy losses--combination and its psychological sequel]. AB - The authors surveyed the literature focused on childhood mistreatment and pregnancy loss. They present definitions and classifications of child abuse (emotional, verbal, physical, sexual), child neglect (physical, emotional intellectual) and pregnancy losses (especially miscarriage and abortion). In the second part of the paper a correlation between abuse and pregnancy loss is displayed as well as a correlation between child abuse and neglect. The different kinds of pregnancy losses are viewed as similar in aspect of psychological sequel, though their intensity and particular character depends on the mother's contribution to the loss of her child. In the last part, the consequences of the above correlation are discussed. The clinical observations suggest, that abuse and neglect experienced in childhood increases the probability of pregnancy loss in adulthood. On the other hand, the loss of on unborn child is one of the factors causing child abuse and neglect. PMID- 12725022 TI - [Group therapy for people damaged by child abuse, neglect and pregnancy loss combination]. AB - The authors reviewed the literature focused on groups for people damaged by childhood mistreatment and pregnancy loss. They also presented II-PLCARR therapy designed for those who suffer from a sequel of abuse, neglect and pregnancy loss combination. Time limited group therapy for people damaged by abuse, neglect and pregnancy loss was designed at the Institute of Pregnancy Loss Child Abuse Research and Recovery, Victoria, Canada. The phases of the program include e.g.: realising mistreatment and damage; realising excuses, denial and resistance; training of assertion; resisting manipulation; accepting partial responsibility; facing guilt, mourning the loss of childhood; passing through despair; dead babies rehumanization and mourning; reconciliation with victims, observers and perpetrators; negotiation of realistic expectations; attenuations of unnecessary pair bonding; learning to love; celebration; good good-byes. The study of IIPLCARR therapy effectiveness is displayed as well. PMID- 12725023 TI - [Psychological interventions and their immune consequences]. AB - Psychoneuroimmunology is a new tendency in science which integrates medicine and social sciences. An interdisciplinary point of view on the etiology and treatment of many diseases fortifies the empirically holistic conception which is very old in medicine. The disease is the result of the collapse of the defensive mechanisms in stress. The nervous, hormonal and immune systems are integrated. The social environment and stress influence the individual's personality and cause diseases, frequently cause immunosuppression. Psychotherapy can "wake" up the immune system. The results of research in psychoneuroimmunology are particular significance for psychosomatic medicine since they explain in a systemic way the earlier clinical observations and scientific studies concerning the influence of stress on the health condition. Studies of the immune system conditioning and the influence of this the relaxation training on this function are very important in coping with the disease. PMID- 12725024 TI - [The importance of community services for rehabilitation and social support to decrease psychiatric hospitalization]. AB - GOAL: The evaluation of community services for social support concerning minimalization of psychiatric hospitalizations. METHOD: Mean duration, global time and the number of stays in hospital ward, day treatment ward and "home hospitalisation"; in two year periods before and after their admission to an Occupational Therapy Workshop (O.T.W.), a Community Home Of Mutual Help (C.H.O.M.H.) or to Community Specialised Social Help Services (C.S.S.H.S.) at home were compared in the group of 73 chronic patients. RESULTS: In two years after the admission to the social support units, the number of all the hospitalizations was decreased (twice or three times). A mean duration of stay in the hospital ward and in the day treatment ward shortened (from 101 days to 48.4 days, from 108 days to 73.3 days), and "home hospitalisation" prolonged (from 112 days to 190 days). The global time of stay in the hospital and in the day ward became significantly shorter (by 12.2% and 5.1%), but "home hospitalisation" did not change significantly. The most effective was the care of specialised services at home connected with the participation at the same time in one of the day units O.T.W. or the C.H.O.M.H. (16 persons). The minimalization of the number (3x) and the time of staying in the day treatment ward referred to the participants of the O.T.W. or the C.H.O.M.H. (by 7.2%, 7.9%), but not of the C.S.S.H.S. at home. CONCLUSIONS: Introducing of O.T.W., C.H.O.M.H. and C.S.S.H.S. resulted in a significant reduction of hospital stays. The best results come from the combination of the care given by the daily units (O.T.W. or C.H.O.M.H.) with specialised services at home. The care supplied at the O.T.W. or the C.H.O.M.H. reduced the number and time of staying in the day treatment ward. PMID- 12725025 TI - [Effect of olanzapine treatment on cognitive functions in patients with schizophrenia]. AB - AIM: The aim of this study was to assess whether olanzapine treatment results in an improvement of cognitive functions that became impaired by the schizophrenic process. METHOD: The correlation of the intensity of schizophrenic symptoms and duration of the disease with the level of cognitive functions were also examined. The schizophrenic patients were examined in three steps: before beginning of olanzapine treatment, after 6 weeks of treatment, and after approximately 12 months since the beginning of the olanzapine treatment, 36 patients were examined (9 female, 27 male) aged 20-53 (X = 25.4). The following tests were applied: Positive and Negative Syndromes Scale--PANSS, Squire's Memory Questionnaire, sub scales: Digit span and Similarities of the Wechsler's Intelligence Scale, Long term memory sub-test of the Choynowski's Memory Scale, Diagnosis of Brain Damages, Clock Completion Test, Decroly's Box. RESULTS: The olanzapine treatment of schizophrenia was characterised by significant antipsychotic efficacy including both positive and negative symptoms. The positive symptoms were associated with the attention range, the planning process context and the immediate memory with the tendency towards loosing remembrance in the process of non-verbal learning. The negative symptoms were associated with concrete conceptual thinking. Improvement of cognitive functions was noted for almost all of the examined functions. The effect of olanzapine therapy in schizophrenic patients may be regarded as personality integrating. PMID- 12725026 TI - [Subjective and objective assessment of memory functions in patients with schizophrenia treated with olanzapine]. AB - Schizophrenic patients often complain of intellectual functioning impairment. The aim of this survey was to define the subjective memory loss by means of objective tests. The tests were carried out in three steps: before the admission to the clinic, after 6 weeks of hospitalisation, and after approximately 12 months from the beginning of the olanzapine treatment, 36 patients were examined (9 female, 27 male) aged 20-53 (X = 25.4). The following tests were applied: PANSS, Memory Questionnaire (Squire, Zouzounis 1988), subscales: Digit span and Similarities of the Wechsler Intelligence Scale, subtest for Long-term memory of the Choynowski's Memory Scale, Diagnosis of Brain Damages (Weidlich, Lamberti), Clock Completion Test, Decroly Box. The subjective assessment of memory functioning improved during the treatment with olanzapine. Before the hospitalisation and after 6 weeks of treatment patients' memory complaints were associated with the visuospatial memory. After 12 month of the olanzapine therapy the memory complaints were associated with audioverbal memory. Age, level of education and objective intensity of psychopathological symptoms did not influence patients' subjective assessment of memory functioning. PMID- 12725027 TI - [Effects of fenvalerate and phoxim insecticides on glutamate and gamma aminobutyric acid immunoreactive cells in the central nervous system of rats]. AB - In order to study the neurotoxic effect and the mechanism of fenvalerate and phoxim insecticides, rats were orally treated with fenvalerate (20 mg/kg BW), phoxim (160 mg/kgBW) and fenvalerate (20 mg/kgBW) plus phoxim (160 mg/kgBW). The glutamate (Glu) and gamma-aminobutyric acid(GABA) immunoreactive cells of the central nervous system of rats were observed by immunohistochemical method. The results showed that the total number, percentage of positive area and integral optical density of Glu immunoreactive cells in cerebral cortex, hippocampus and striate exhibited a characteristic decrease in the groups fed with fenvalerate and fenvalerate plus phoxim, while those of GABA were enhanced to certain extents. No obvious changes in Glu or GABA were observed in the phoxim group as compared with the controls, and no significant difference was observed between the fenvalerate group and the fenvalerate plus phoxim group. The results suggest that the disturbance of Glu and GABA metabolism may play an important role in the development of pyrethroid neurotoxicity, and there is no interaction between phoxim and fenvalerate pesticides on Glu and GABA activities. PMID- 12725029 TI - [K-ras gene mutations of asbestos and welding-fumes related human lung cancer]. AB - To investigate the K-ras gene mutations in asbestos and welding-fumes related human lung cancer (hLC), comparing with the mutational spectrum of K-ras gene in non-occupational hLC, 8 cases of non-occupational hLC, 9 cases of asbestos related hLC and 4 cases of welding-fumes related hLC were collected. The techniques of genomic DNA extraction, nested-polymerase chain reaction (PCR), polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP), polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) and DNA direct sequencing were employed. THE RESULTS: showed the mutation frequencies of K-ras gene were 25% in non-occupational hLC and welding-fumes related hLC, whereas 44.4% in asbestos-related hLC. Mutations mainly occurred in codons 12 and 15. A mutation of codon 13 was detected in welding-fumes related hLC. The type of base substitution was principally G-->T transversion in non-occupational hLC, whereas mainly G-->A transition in asbestos and welding-fumes related hLC. THE RESULTS suggested there were obvious differences of K-ras gene mutations among non-occupational hLC, asbestos and silica related hLC. Therefore, different mechanisms of carcinogenesis were implicated among them. However, no speific spectrum of K-ras gene mutation can be derived from welding-fumes related hLC because of the limitation of samples. PMID- 12725028 TI - [Effects of phoxim on sperm production and motility of rats]. AB - The effects of phoxim at various dosage levels on sperm production and motility in rats were studied. Sperm motility was measured by computer assisted sperm analysis (CASA). The results showed that rats exposed to phoxim at higher doses(24.5 and 73.5 mg/kgBW) for 60 successive days showed significant reduction of daily sperm production as compared with the control (P < 0.05, P < 0.01). Sperm motion parameters such as currilinear velocity (VCL), straight line velocity(VSL), beat cross frequency(BCF), linearity(LIN) and straightness(STR) were significantly decreased than those of the control(P < 0.05, P < 0.01) which indicated a dose-dependent relationship to some extent. However, there was no obvious change in body weight, testis weight and the rate of motile sperm in epididymis. It suggests that the male reproductive toxicilIty of phoxim at higher doses (24.5 and 73.5 mg/kgBW) mainly induces a reduction of sperm production and motility. PMID- 12725030 TI - [Effects of local vibration on the levels of plasma endothelin and nitric oxide in rabbits]. AB - In order to explore the effects of local vibration on plasma endothelin(ET) and nitric oxide (NO) and the relations with vibration-induced vascular impairment(VVI), plasma ET and NO levels of rabbits were measured after exposure to different vibration power and different exposure period. The results showed an increasing tendency in plasma ET levels, and a decreasing tendency in plasma NO levels in experiment groups, while lasting vibration exposure and enhancing vibration power. It suggested that local vibration could result in the increasing of plasma ET level and decreasing of plasma NO level, which might be one of the mechanisms of vibration-induced vascular impairment. PMID- 12725031 TI - [Combined effects of lead and ethanol on rat sperms]. AB - In order to study the combined effects of lead and ethanol on rat sperm, 54 male rats were divided into 9 groups randomly. One group was given distilled water as control, and the others (A-H) groups were given lead and ethanol at the concentrations of lead 11 mg/kgBW and lead 22 mg/kgBW, ethanol 0.9 g/kgBW, ethanol 1.8 g/kgBW, lead 11 mg/kgBW + ethanol 0.9 g/kgBW, lead 11 mg/kgBW + ethanol 1.8 g/kgBW, lead 22 mg/kgBW + ethanol 0.9 g/kgBW, lead 22 mg/kgBW + ethanol 1.8 g/kgBW in stomachs respectively for four weeks. The rats were sacrificed by the end of the fourth week. Epididymis were isolated and examined for sperm motility, counts and morphology. The results showed that the spermatozoa motility as well as spermatozoa count was lower; the spermatozoa morphological changed and blood lead level and ratio of epididymis to weight were higher in all combined experimental groups than those in groups with lead or ethanol alone. ANOVA analysis indicated that there were interaction between lead and ethanol on above indices (P < 0.05 P < 0.01). The multiple linear regression showed that there was linear correlation between blood lead and spermatozoa motility, spermatozoa morphological change. It is concluded that there were notable combined effects of lead and ethanol on semen quality in male rats. PMID- 12725032 TI - [Ethanol-induced apoptosis in human HL-60 cells]. AB - The purpose of this study was to examine the effect of ethanol on apoptosis of human promyelocytic leukemia cell line HL-60 cells and search for a possible reason for reduced PMNs, monocytes, lymphocytes in the blood of alcoholics. Human promyelocytic leukemia cell line HL-60 cells was used as a model of apoptosis, with ethanol being the interfering factor. Qualitative and quantitative detection was done by both electron microscopy and flow cytometry. Compared with the control treatment of HL-60 cells with ethanol at 50, 100, 150, 200, 250 mmol/L for 3 days increased apoptotic cells significantly (P < 0.01), and apoptosis was dose-dependent for ethanol. It suggest that ethanol can induce apoptosis of HL-60 cells. So as far as reduced PMNs, monocytes, phagocytes and lymphocytes of alcoholics are concerned, ethanol-induced apoptosis may well be an important reason. PMID- 12725033 TI - [Study on the effect of alcohol on embryonic development by using in vitro post implantation rat whole embryo culture]. AB - In order to explore the effects of drinking alcohol during pregnancy on embryonic development and its mechanisms, a post-implantation whole embryo culture(WEC) technique was used. The 9.5 day rat embryos were explanted in rat serum medium(immediately centrifugal serum, ICS) with alcohol(0.0.4.1.0, 2.00 and 4.00 g/L), and cultured for 48 hours. The index of embryo development and morphological scores induced by alcohol were observed. The result showed that alcohol had obviously effects on the development and growth of embryos with a dose-response relationship. Embryonic development of 0.4 g/L group was not significantly different from the control group, whereas 1.0 g/L group could interfere with the development score of mid-brain, forebrain, neurotube, and visceral yolk sac(VYS) circle obviously. All scores of the 2.00 g/L group were significantly lower than that of control group (P < 0.05). Moreover, the rate of embryo lethality and teratogenecity were obvious increased. It is concluded that alcohol has developmental toxicity and teratogenicity. The target organ affected by alcohol is brain. The effects of alcohol on the developmental differentiation of visceral yolk sac and DNA synthesis are probably related to its developmental abnormalities. PMID- 12725034 TI - [Study on antagonistic effects of selenium and zinc on the renal impairments induced by fluoride in rats]. AB - Wistar rats were provided with distilled water containing NaF(100 mg/L), and were administered through gavage with Na2SeO3[0.1 mg/(kgBW.d)] and/or ZnSO4[14.8 mg/(kg BW.d)]. The results of biochemical, pathological and ultrastructural examinations showed that fluoride could cause serious renal impairments. The major damage induced by fluoride was epithelia of proximal renal tubules. The lipid peroxidation might be one of the mechanisms of fluoride toxicity. Na2SeO3 and ZnSO4 could antagonize the renal impairments induced by fluoride through their antioxidation. The cooperative effect of Na2SeO3 and ZnSO4 was more powerful than either Na2SeO3 or ZnSO4 alone. PMID- 12725035 TI - [Study on the molecular mechanism of genetic damages in body cells in mice treated with organic mutants in drinking water]. AB - The genetic toxicity of some volatile compounds of chlorinated by-products have been determined in animal studies. But little is known at present about the genetic toxicity of nonvolatile matter in drinking water in vivo assay. In a sub acute experiment, the mice were exposed to the organic compounds extracted from in-home tap water. The pathologic changes in the liver and kidney tissues were observed under microscope and electron microscope. The frequencies of micronuclei in polychromatic erythrocytes in mice chest bone marrow in the three treated groups were tested. The point mutants in p53 gene of liver, kidney and colon tissues in the high dosage group were examined by PCR-SSCP. Especially, the frequencies of mutations in exon 7 of p53 gene in those mice were higher than those in exon 5 of p53 gene. It showed that the study on the molecular genetic toxicity of organic mutants in vivo assay was possible. PMID- 12725036 TI - [Effects of microwave radiation on lipid peroxidation and the content of neurotransmitters in mice]. AB - Male LACA mice were exposed to 900 MHz microwaves at the power densities of 0,1,2 and 5 mW/cm2. The resulting specific absorption rates(SARs) were 0, 0.22, 0.44 and 1.10 W/kg respectively. Animals were exposed at the dosage of 1 hour a day for 35 days. The results showed that the activity of glutathione peroxidase (GSH Px) increased and the concentration of malondialdehyde (MDA) decreased in blood, and the concentration of noradrenaline (NE) increased in brains in the microwave exposed groups at 1 mW/cm2 compared with the sham-exposed group. But no significant changes were found in the microwave exposed groups at 2 and 5 mW/cm2. No significant changes of the concentrations of 5-hydroxytryptamine(5-HT), 5 hydroxyindole acetic acid(5-HIAA) and dopamie(DA) in brains were found. PMID- 12725038 TI - [Monitoring on blood lead and cadmium in Nanjing inhabitants from 1983 to 1998]. AB - Blood lead (Pb) and cadmium (Cd) of residents in Nanjing were monitored from 1983 to 1998. The results showed that the blood Pb concentration was male > female, smokers > nonsmokers. Blood Pb in female nonsmokers increased gradually during recent ten years. Blood lead level was also significantly different between urban residents and suburban residents in 1998. Blood Cd concentration in female nonsmoker < male nonsmokers < male smokers < female smokers, male smokers > male nonsmokers. The blood Cd of female nonsmokers kept steady in the last ten years. The concentration of blood Cd of urban residents was accordant with suburban residents in 1998. PMID- 12725037 TI - [Effect of typical aldehyde pollutants on the DNA damage of spleen lymphocytes of mice]. AB - The DNA damage of spleen lymphocytes induced by three typical aldehyde pollutants was studied in vitro with microgel single cell electrophoresis assay (comet assay). It showed that the comet cells induced by the three chemicals were observed, and the dose-dependent relations were existed. It suggested that the three aldehydes could induce DNA strand breakage. PMID- 12725039 TI - [Effects of cadmium chloride on the malignant transformation of human embryo lung fibroblasts]. AB - The malignant transformation of human embryo lung fibroblast (HELF) induced by CdCl2 was tested by 3H-TdR incorporation, chromosome analysis and flow cytometry. Chromosome aberration, the ratio of cells in different cell cycle phases and the quantities of DNA synthesis of different groups were determined. The results showed that the morphology of HELF changed in three levels of cadmium chloride (I 0.004 mumol/L, II 0.02 mumol/L and III 0.04 mumol/L) treated for three times. The cell growth was disordered and overlapping without density regulating effect and the transforming foci were formed. The quantities of chromosome were decreased or increased and the dicentric and acentric fragments were found in experimental groups. The ratio of cells in S phase increased and the quantities of DNA synthesis in experimental groups were 2.4-3.8 times higher than that of negative control group. The chromosome aberration of cells, the increase of cells in S phase and the increase of DNA synthesis may play some roles in the malignant transformation of HELF induced by CdCl2. PMID- 12725040 TI - [Determination of volatile organic compounds in blood by headspace solid-phase microextraction-gas chromatography]. AB - The headspace solid-phase microextraction (HS-SPME) is a novel extraction technique and has been developed rapidly. It is a fast, simple, solventless and sensitive method for sampling, separating, extracting, injecting and analyzing volatile organic compounds. This paper presents the research work in detecting volatile organic compounds(including ten compounds) in blood. The extraction fiber is made by fused-silica fiber with 100 microns polydimethylsiloxane (PDMS). The extraction time of the method was 10 min. The thermal desorption time was 1 min. It was found that the optimized location of the extraction fiber in the injector of GC was to put the whole needle in the injector. The precision of the method was determined to be less 5% relative standard deviation (RSD). The linear range of the detection was rather wide. The lowest detectin limits (LODs) were all < or = 5 ng/ml. PMID- 12725041 TI - [Effects of glutamate and glutamine on learning and memory of rats]. AB - The effects of glutamate (Glu) and glutamine (Gln) on learning and memory of rats and the mechanism involved were examined. Three groups of rats were fed with stock diet (control group), and the diets supplemented with 2% Glu (Glu group) or 2% Gln (Gln group) respectively. Compared with control group, the active avoidance response rats of Gln group in shuttle box test was significantly elevated while the time of passive avoidance response shortened. The contents of brain free amino acids, such as Gln, Glu, aspartic acid and arginine, activity of brain nitric oxide synthase (NOS), Bmax of N-methyl-D-aspartate(NMDA) receptors in hippocampus of Gln group were all increased significantly. Such changes were not observed in Glu group. The results suggested that Gln could improve the learning ability and memory of rats, which might be related to the change of amino acids composition of brain, the synthesis of messenger nitric oxide and the binding capacity of NMDA receptor in hippocampus. PMID- 12725042 TI - [Effect of homocysteine on the proliferation and differentiation of rat embryo midbrain cell]. AB - In order to determine the potential mechanisms by which HCY may exerts its teratogenic effects, a micromass culture for rat embryo midbrain cell was used to reveal the differentiation and proliferation of the cell under different HCY doses (0-10 mmol/L). The results showed that with increased concentration of HCY, the promotion effect changed into inhibition. The concentration inhibiting the formation of differentiated foci by 50% of the control was 4.3 mmol/L, and 2.3 mmol/L when S9 was added. It had an inhibition effects on proliferation in the testing scope. The result suggested that HCY obviously inhibited more cell differentiation than proliferation. The teratogenic hazard of HCY may play an important role in the occurrence of neural tube defects. PMID- 12725043 TI - [Effect of fish oil supplementation on fatty acid composition and neurotransmitters of growing rats]. AB - In order to observe the change of brain fatty acid composition and neurotransmitter by DHA supplementation during the development stage of young rats, pregnant rats were randomly divided into four groups: normal control group (O), low (L), medium (M) and high (H) dose-DHA supplemented groups. The fatty acids composition, the levels of 5-hydroxy-tryptamine (5-HT), dopamine (DA), norepinephrine (NE) and somatostatin (SS) in ceretral cortex and hippocampus of offsprings were measured. The body weight of pregnant rats and offsprings were also observed. The results showed that DHA content of brain homogenate and the levels of 5-HT, DA and SS in hippocampus of experimental young rats were significantly higher than those of the control group. And the body weight of young rats in the three DHA-supplemented groups were retarded in different degree. It was concluded that DHA supplementation changed the fatty acid composition and increased the levels of 5-HT, DA and SS in hippocampus. Proper ratio of n-3 and n-6 fatty acids in dietary supplementation should be considered. PMID- 12725044 TI - [Study on genotypes of cystathionine beta-synthase in neural tube defects]. AB - Mildly elevated maternal plasma homocysteine (Hcy) levels (hyperhomocysteinemia) have recently been observed in some neural tube defects(NTDs) pregnancies. The aetiology of NTDs is also known to have both genetic and nutritional components. The frequency of two relatively common mutations in the enzyme of cystathionine beta-synthase (CBS), which is one of the main enzymes that controls Hcy level, was examined. Among 76 nonpregnant women in the study, 40 of whom had given birth to infants with NTDs and 36 control women previously had normal offspring. The results showed that no significant difference was found between NTDs mothers and non-NTDs mothers for the frequency of T833-C and the G919-A mutations in the CBS gene. The data suggests that the mother's genotype is not the direct factor involved in the pathogenesis of NTDs. PMID- 12725045 TI - [Zinc modulates the Ca(2+)-ATPase activity of rat osteoblast membrane]. AB - The effect and mechanism of zinc on Ca(2+)-ATPase and Na+, K(+)-ATPase activity of rat osteoblast membrane were studied. The activities were measured by colorimetric method. The effect of special inhibitors of protein kinase C or calmodulin on membrane Ca(2+)-ATPase and Na+, K(+)-ATPase activity which induced by zinc were also examined. The results showed that zinc could increase membrane Ca(2+)-ATPase activity, but no influence was observed on Na+, K(+)-ATPase activity. R24571, a special inhibitor of calmodulin, had no effect on membrane Ca(2+)-ATPase activity. Calmodulin also could not promote the basic Ca(2+)-ATPase activity of membrane. But trifluoperazine, a special inhibitor of protein kinase C, obviously reduced its activity. It was concluded that the modulation of zinc on membrane Ca(2+)-ATPase activity of rat osteoblast could be mediated by protein kinase C. PMID- 12725046 TI - [Effects of tea on aberrant crypt foci and colorectal tumors in rats]. AB - The present study was designed to investigate the chemopreventive effects of green tea and tea pigments (the main component of black tea) on 1,2 dimethylhydrazine (DMH)-induced rat colorectal carcinogenesis. Compared with the positive control group, green tea and tea pigments groups had less numbers of aberrant crypt foci (ACF) (P < 0.01) at the end of week 16. At the end of week 32, all rats in the positive control group developed colorectal tumors with an average of 2.6 tumors per rat, and a mean volume of 294.7 mm3 per tumor, while in the groups drinking green tea and tea pigments, the average numbers of colorectal tumors per rat were only 47.1% and 43.1% of controls respectively, and the mean tumor volume was inhibited by 77.1% and 68.1% respectively. It was concluded that tea pigments had a chemopreventive effect on colorectal tumor and ACF formation can be used as a useful intermediate end point to study chemopreventive effects on colorectal cancer. PMID- 12725047 TI - [Acute toxicity and bioavailability of nano red elemental selenium]. AB - The reaction ratio of nano red elemental selenium (nanose) with GSH in vitro was one-tenth of that of sodium selenite. Mice were fed with nanose and sodium selenite separately at 50 micrograms/kg BW for 30 days. Both selenium forms could significantly increased Se concentration of blood, liver and activity of blood GSH-Px. Acute toxicity experiment of mice implied that the acute toxicity of nanose was nearly one-seventh of that of sodium selenite. The LD50 for nanose se was 112.98 mg kg BW, and the LD50 for sodium selenite was Se 15.72 mg/kg BW. The acute toxicity of nanose was low for mice. PMID- 12725048 TI - [Establishment of monoclonal antibodies against aflatoxin M1]. AB - The monoclonal antibodies against aflatoxin M1 were established. Spleen cells from Balb/c mice immunized with AFM1-oxime-BSA conjugate were fused with murine Sp2/0 myeloma cells. Three hybridoma cell lines secretine monoclonal antibodies against AFM1 were established after the fusion cells subcloned for 3-4 cycles. These antibodies were disignated as 3G2, 3G3 and 6G8, respectively. All of them belonged to the subtype of IgG1. The titres of the antibody in ascites ranged from 1:3.2 x 10(6)-1:20 x 10(6). There was no cross reaction between 6G8 monoclone antibody and other types of aflatoxin. PMID- 12725049 TI - [Weights in Horvitz-Thompson statistic for complex samples]. AB - The problem of using ordinary statistical methods basically for simple random samples in analyzing data from complex surveys for non-simple random samples was discussed. Horvitz-Thompson statistic and the poststratification weights were introduced. Illustrative examples were given to show the validity and superiority of Horvitz-Thompson statistic in comparison with statistic used for simple random samples. PMID- 12725050 TI - [Zinc protective effects on pig retinal pigment epithelial cell damage of lipid peroxide induced by 2450 MHz microwave]. AB - To observe the lipid peroxide damage in the cultured pig retinal pigment epithelial cell(RPE) induced by microwave and the protective effects by zinc (Zn). RPE cells were divided into 3 groups in terms of microwave intensity(the group of 10, 20 and 30 mW.cm-2) and each group was exposed to microwave for 1 hour. After adding ZnSO4 into DMEM culture fluid, the group of 30 mW.cm-2 was exposed to microwave. The activity of SOD and the content of MDA were measured. The results showed that the activity of SOD decreased significantly compared with the control group(P < 0.001). The content of MDA increased significantly compared with the control group(P < 0.001). Administration of Zn alleviated the increase of MDA and decrease of SOD. In the experiment, 2450 MHz microwave induces the lipid peroxide damage in RPE cells. Zn can enhance the antioxidation ability of cells and alleviate the damage to some extent. PMID- 12725051 TI - [Role of interleukin-6 and platelet-activating factor in delaying neutrophil apoptosis]. AB - In the present study we investigated the role of platelet-activating factor (PAF) and interleukin-6 (IL-6) in delaying polymorphonuclear neutrophil (PMN) apoptosis. Isolation of PMN was performed by using the discontinuous plasma Percoll gradient technique. PMN was cultured in enriched RPMI 1640 media at 2 x 10(7) PMN/ml for 24 h. Subgroups were treated with IL-6 or PAF or pretreated with PAF receptor antagonist BN52021 before IL-6 or PAF were added. Morphological assessment and quantitation of apoptosis were performed with acrodine/ethidium bromide stain and epifluorescent microscope. The results showed that both IL-6 and PAF suppressed PMN apoptosis. Pretreatment of PMN with BN52021 abrogated the effects of IL-6 and PAF. It is suggested that PAF may be a crucial cytokine in the suppression of PMN apoptosis. These observations may contribute to elucidating the mechanisms of IL-6 and PAF in mediating postinjury hyperinflammation and secondary organ dysfunction and provide a clue for the prevention and treatment of the conditions. PMID- 12725052 TI - [Effect of cadmium on apoptosis of spermatogenic cells of rat testis and the protection effect of zinc against it]. AB - To study the effect of cadmium on the apoptosis of spermatogenic cells of rat testis and the protective effect of zinc against it. 24 Wistar male rats were divided into 3 groups (cadmium, cadmium and zinc, and control rats). Rats were injected with low-dose CdCl2 (2 mg/kgBW), zinc acetate(ZnAc, 15 mg/kgBW) before and after injected with low-dose CdCl2 and ZnAc(50 mg/kgBW). The control rats were injected with same-dose of 0.9% NaCl. After 7 days, the apoptosis of spermatogenic cells was studied with in situ nick translation(ISNT) technique. Compared with control group, the number of apoptosis spermategenic cells was obviously increased in cadmium group(P < 0.01, t = 3.87), but it was not significantly different in cadmium and zinc rats. Cadmium treatment could accelerate testis apoptosis. Zinc could proven it. PMID- 12725054 TI - [Toxicity of anti-herbicide gene (BAR) transgenic rice]. AB - In order to evaluate the safety of anti-herbicide gene(BAR) transgenic rice, acute toxicity experiments, mutation experiments and a 30-day feeding test were conducted. The results were as follows: The oral LD50 for mice and rats was larger than 21.5 g/kg BW and no mutation was found. The rats consuming 16.32 and 64 g/kg BW were in good growth and development at the 30-day feeding test. The body weight, food utilization, routine blood tests, the ratio of organ weight to body weight, and patho-histological observations had no obvious change. The none effect dosage for the transgene rice was 64 g/kg. PMID- 12725053 TI - [Toxic effect of fluoride-arsenic on the reproduction and development of rats]. AB - To determine the effects of the fluoride-arsenic exposure on the reproductive function and the development of offspring. Two generations-one nest reproduction was studied in Wistar rats to assess the changes of reproductive function and structure of ovary and development of their offspring after rats were exposed to different doses of fluoride(NaF)-arsenic(As2O3) orally. The rates of the pregnancy, normal parturition, survival and feeding survival in high dose group were lower than that in low dose groups and control. The rates of pregnancy were 73.08% and 61.54% in high dose groups, and 92.31% and 84.62% in the control. But the contents of fluoride and arsenic in rats' offspring were increased. The weights of rats' offspring in high dose group were lighter than that in control. The average weights were 38.45 g in high dose group and 60.84 g in control one month after birth. Pathological changes could be found in each stage of oocytes with transmission electron microscope(TEM). Conclusion Fluoride-arsenic exposure has adverse effects on the reproductive system and the development of rats' offspring. PMID- 12725055 TI - [Toxic effect of organic extracts of drinking-water on the culture luteal cells of rats]. AB - A separation method of rat luteal cells was established. The results show that the survival rate of the cells is more than 95% during the whole operation. 14-20 days of pregnancy, 10(6) cells/ml and 4 hours of culture are the suitable conditions. The organic extracts of drinking-water produced significant dose related increases(maximally three-fold) in progesterone concentration in the culture medium. Thus, the effects produced by the organic extracts of drinking water in vitro may reflect the damage to the luteal cells. PMID- 12725056 TI - [Contrast test using three kinds of air-microorganism samplers LWC-I, CA6 and CA2 in public places]. AB - A sampling experiment was conducted using three kinds of air-samplers, i.e. LWC 1, CA6 and CA2, on bacteria and fungus in the air under the same conditions and at the same time yet in three different public places. The results showed that there was no significant difference in the total number of bacteria and fungus colony forming units in the air in different public places using the above mentioned three air-samplers. In comparison, the LWC-1 air-sampler demonstrated a feature of small size, light weight, simple structure, and little noise, therefore, it was easy in carrying and requires no external power source. Such being the case, the LWC-1 air-sampler was more suitable for monitoring microorganism in the air in public places. PMID- 12725057 TI - [Effect of smoking on the guanylin in the injured lung of rats]. AB - In order to survey the influence of smoking on the level of guanylin in lung tissue in rats, a method of passive smoking was used to set up chronic bronchitis model in rats. The content of guanylin was measured by radioimmunoassay. In the smoking group, remarkable pathological changes in the lung tissue were found. There was no significant difference in the contents of guanylin in the plasma of experimental rats, compared with the control(P < 0.05). But in the lung tissue, the difference was significant(P < 0.01). PMID- 12725059 TI - [Rapid determination of the ozone in water]. AB - A new potassium iodide--N, N-diethy1-1,4-phenylen-ediammonium sulfate (DPD) spectrophotometry method for the determination of the ozone in water was established. The method was based on the quantitative reaction between the ozone and potassium iodide in buffered solution(pH 3.35): O3 + 2I- + 2H+-->O2 + I2 + H2O. Iodine could react with DPD regent and produce pink color. Its absorption was determined at 510 nm with spectrophotometer. The range of direct determination was 0-1.00 mg/L. Beer's law was obeyed in this range. The detection limit was 0.01 mg/L. The relative standard deviations were from 1.2% to 2.7% (n = 10). The recoveries were from 98.6% to 106.6%. Water samples were determined with both this method and iodimetry. The results were comparable. The method was applied to the determination of the ozone in water sample with satisfactory results. It was practical and reliable. PMID- 12725058 TI - [Dynamic study on blood and milk lead levels of pregnant women in three districts of Hubei]. AB - In order to study the effect of lead exposure on the health during the period of pregnancy, 96 pregnant women and 105 non-pregnant women with no known occupational exposure to heavy metals from urban area(Wuhan), iron mining area(Daye) and rural area(Yidu) were selected. Blood and milk samples were collected under strict quality control and analyzed by flame atomic absorption spectrometer. The general status of subjects were collected by a medical interviewer. The results showed that the blood lead in the non-pregnant women were higher in the urban area, but the blood lead in the pregnant women and the milk (colostrum and mature milk) lead were lower in the urban area. It is concluded that the risk of exposure to low concentration of lead during pregnancy must not be ignored. PMID- 12725060 TI - [Determination of trichlosan in cosmeticy by high-performance liquid chromatography]. AB - A method was developed for the determination of an antiseptic-trichlosan in cosmeticy by HPLC followed by diode-array detection. The chromatographic separation was performed on C18 column with a mobile phase of acetonitrile water(60: 40, V/V) which was adjusted to pH3.0 by glacialacetic acid. The UV wavelength was 280 nm. Retention time as well as UV spectrum were used for the qualitation of trichlosan, and peak area or peak height for quantitation. The method showed good accuracy and precision average recovery of trichlosan from different cosmetics was between 91.86% and 98.83%. Relative standard deviation was between 0.72% and 2.75%. PMID- 12725061 TI - [Mechanism of impairment to microtubule polymerization resulting from zinc deficiency during pregnancy and lactation in mice]. AB - To probe into the mechanism of zinc deficiency on microtubule polymerization impairment, the learning ability and the levels of alpha-tubulin, beta-tubulin and microtubule-associated protein 2 expression in the brain of zinc deficient offsprings, maternal ICR mice were fed with experiment diets containing different levels of zinc(1, 5, 30 and 100 mg/kg) during pregnancy and lactation respectively. On the postnatal day 70 of offsprings, the learning ability and the expression of alpha-tubulin, beta-tubulin and microtubule-associated protein 2 in the brain were examined by shuttle box and Western blot assays respectively. The results showed that the number of trials needed to reach the learning criterion for zinc deficient groups (1 and 5 mg/kg) was much higher than that for non zinc deficient groups (30 and 100 mg/kg). The levels of alpha-tubulin, beta-tubulin and microtubule-associated protein 2 expression in the brain of zinc deficient offsprings (1 and 5 mg/kg) were lower than those in the brain of offsprings whose dams fed with zinc adequately supplied diet (30 mg/kg) and zinc supplemented supplied diet (100 mg/kg) respectively. These results suggested that the inhibition of alpha-tubulin, beta-tubulin and microtubule-associated protein 2 expression might be the most important mechanism of microtubule polymerization decline resulting from zinc deficiency, which had close relationship with brain function impairment. PMID- 12725063 TI - [Effects of dietary lipids on serum cholesterol of elder hypercholesterolemic patients]. AB - The intake of dietary lipids of 167 elder hypercholesterolemic patients was collected by a three-day recorded dietary survey. At the mean time, their serum total cholesterol (TC), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) levels were detected and analyzed. The results showed the effects of dietary lipids and body mass index (BMI) were great on serum cholesterol levels; BMI, dietary cholesterol and saturated fatty acid were positively related to serum TC and LDL-C; serum HDL-C and HDL-C/TC ratio were positively related with dietary monounsaturated fatty acid, and negatively related with BMI. The results indicate that decreasing the intake of dietary cholesterol and saturated fatty acid, controlling body weight and increasing dietary monounsaturated fatty acid intake will have important benefits on the prevention and treatment of hypercholesterolemia and cardiovascular diseases. PMID- 12725062 TI - [Effect of tea polyphenols and tea pigments on inducing the activity of phase II detoxicating enzymes and on the chemoprevention of liver precancerous lesions]. AB - In order to elucidate the chemoprevention mechanism of tea, the regulative effect of tea polyphenols and tea pigments on phase II enzyme, glutathions S-transferase (GST) activity and its isoenzymes in Wistar rat were studied. Rat liver precancerous lesions were initiated with diethylnitrosamine (DEN), promoted with partial hepatectomy and carbon tetrachloride. The rats were killed after 8 weeks of treatment. Liver cytosol GST activities were tested. The GST 1-1, 1-2, 3-3 and Pi isoenzymes were detected with Western Blot method. The results showed that the activities of GST increased greatly in the tea polyphenol and tea pigment groups, while the activities were decreased in positive control group. Both tea polyphenold and tea pigments increased the activities of classes GST alpha (GST1 1, 1-2) and GST mu(GST3-3). The increase of GST alpha was more in tea polyphenol group, and the increase of GST mu was more in tea pigment group. Both tea polyphenols and tea pigments inhibited the over expression of GST Pi in the rats treated with DEN/PH/CCl4. The study indicated that tea polyphenols and tea pigments modulated phase II enzymes effectively, thus inhibited the occurrence and development of the precancerous lesions of rat liver. PMID- 12725064 TI - [Effects of vitamin A on Hox 3.5 gene expression in mouse embryo]. AB - In order to investigate the effects of vitamin A(VA) deficiency and VA supplementation on Hox 3.5 gene expression in mouse embryo, 60 Kunming weaning female mice were divided into four groups. The normal control group (N) was fed diet containing VA 4,000 IU/kg. The other 3 groups were fed VA deficient diet (VA 0 IU/kg diet) for 16 weeks and then make all mice mating with normal male mice. Group A was fed VA deficient diet all through the experiment. Group B and group C were fed diet supplemented with VA (VA 10,000 IU/kg) from the day zero and from the 7th day after conception respectively. On the 12th day of gestation, 6 pregnant mice in each group were killed and the embryos were taken immediately. In situ hybridization was used to determine the contents of mRNA of Hox 3.5 gene in mice embryos by digoxin-labeled cDNA probes. The results revealed that the contents of Hox 3.5 gene mRNA in the embryos of group A and C decreased significantly compared from group N and B, and that in group B had no difference from group N. The mRNA contents of group C were higher compared with group A, but still was lower than that of group N. It was concluded that vitamin A might influence the development of embryos by regulating the expression of homeobox gene. PMID- 12725065 TI - [Effects of sodium selenite on the activity of GSH-Px and the life-span of Drosophila]. AB - In order to investigate the effects of sodium selenite on the activity of glutathione peroxidase and the life-span of drosophila melanogaster, newly enclosed flies were divided randomly into 4 groups and fed with mediums containing different concentrations of sodium selenite. The activity of GSH-Px was determined by 5,5'-dithionbis (2-nitrobenzoic acid) (DTNB) photometric method. The life-span of drosophila was calculated after survival test. The results showed that the activity of GSH-Px in the female and male flies were increased by the increase of sodium selenite in the medium (P < 0.05). The average life-span and the average maximum life-span of flies in selenium groups were increased significantly as compared with control group (P < 0.05). The response of GSH-Px activity and life-span of male flies to selenium intake was better than that of females. PMID- 12725066 TI - [Effect of calcitonin on the proliferation and membrane fluidity of human osteoblast-like cells]. AB - The effects of calcitonin (CT) on the proliferation and membrane fluidity of human osteoblast-like cells line OS-732 were compared with the effect of 1,25(OH)2D3. The rate of DNA synthesis, cell proliferation, cell cycle and membrane fluidity of OS-732 cells were observed. Results showed that 1IU/ml CT stimulated cell proliferation and DNA synthesis. The percentage of cell in the G2 + M phase was increased and the percentage of cell in the synthetic phase was decreased. The membrane fluidity of OS-732 cells was increased after 0.1 mumol/L and 0.01 mumol/L 1,25(OH)2D3 treatment and was decreased after 1IU/ml CT treatment. It is concluded that calcitonin might increase the cell proliferation and decrease the membrane fluidity. PMID- 12725067 TI - [Effects of black and red rice on the formation of aortic plaques and blood lipids in rabbits]. AB - In order to observe the antiatherosclerotic effects of black and red rice and to find out their mechanism, 24 New Zealand male white rabbits (average body weight 1.91 kg) were divided randomly into three groups (white rice, black rice and red rice groups). The rabbits were fed a cholesterol-enriched diet (cholesterol 5 g/kg, lard 35 g/kg) containing 300 g/kg white, black and red rice powder respectively. The rabbits were sacrificed 10 weeks later. The aorta was dissected and stained in 0.5% oilred-isopropanol solution. The plaque area and total area of aorta were measured by an automatic image analyzer. Serum lipids were determined by an automatic analyzer. The results showed that the plaque area (% of total surface) in the black and red rice groups was significantly lower than that in the white rice group (P < 0.05). Serum TG, TC, LDL-C, ApoB and ApoAI/ApoB were not significantly different among the three groups (P > 0.05). However, the concentrations of HDL-C and ApoAI were significantly higher in the black and red rice groups than those in the white groups (P < 0.05). No significant difference was found between the black and red rice groups. It is concluded that black and red rice might be effective in reducing atherosclerotic plaques on the aorta of rabbits fed a cholesterol-enriched diet. The effectiveness of black and red rice against atherosclerosis might be related to the high level of serum HDL-C and ApoAI. PMID- 12725068 TI - [Inhibition of human gastric carcinoma cell growth by vitamin E succinate]. AB - The effects of vitamin E succinate (VES) on the growth of human gastric carcinoma cell (SGC-7901) were examined and compared with the effects of alpha-tocopherol by MTT assay. It was found that VES(5, 10 and 20 micrograms/ml) inhibited cell growth significantly, but alpha-tocopherol did not show any inhibition effects. The results showed that the anticancer effect of VES and the anticancer mechanism of VES is different from alpha-tocopherol. PMID- 12725069 TI - [Mutation of p53 and Ki-ras gene in human fetal lung fibroblast cells in vitro by sterigmatocystin]. AB - To explore the carcinogenic effects of sterigmatocystin(ST), one of the predominant contaminating mycotoxins in high risk areas of cancer in China, mutation of tumor suppressor gene p53 and oncogene Ki-ras in human fetal lung cells in vitro induced by ST was studied using cell culture and silver-staining PCR-SSCP methods. The results showed that, within 22 weeks of ST treatment, no abnormalities were found for both p53 and Ki-ras gene in electrophoresis. Abnormal electrophoretic migration bands were seen at exon 8 of p53 gene and Ki ras gene in ST-treated human lung fibroblast cells 22 weeks after ST treatment. Thus, the results further confirmed the carcinogenic effects of ST on human fetal lung tissue. PMID- 12725070 TI - [Immunological study on the antitumor effects of fungus polysaccharides compounds]. AB - Fungus polysaccharides compounds (FPC) are the mixture of procyanidins oligomers, glycyrrhetinicacid and polysaccharides of hericium erinaceus, lentinus edodes and poria cocos. The antitumor effects of FPC and its immunity regulating effects as an immunostimulant on the mice burdened with sarcoma 180 (S-180) were studied. FPC (100, 200 and 400 mg/kg BW) was gavaged to mice for 31 days. S-180 was transplanted to these mice on the 21th day. Lentinus edodes group was gavaged 200 mg/kg BW saccharine of lentinus edodes. The results showed that FPC could inhibit the growth of S-180 effectively. The inhibitory rates were 37.74%, 44.73% and 48.32% respectively. The antineoplastic activity of FPC (200 mg/kg. BW) was more effective than polysaccharide of lentinus edodes at the same dose. In S-180 burdened mice, the percentage of L3T4 and the ratio of L3T4/Lyt-2, NK activity and the induced IL-2, IFN-gamma levels were decreased significantly compared with the normal control group. As an immunostimulant, FPC could increase the percentage of L3T4 and the ratio of L3T4/Lyt-2 in S-180 burdened mice, but had no significant effects on the percentage of Lyt-2. Polysaccharide of lentinus edodes alone could also increase the immunity competence of mice burdened with S-180, but was not better than that of FPC at the same dose. It could be concluded that the compound of antineoplastic component could be synergetic. PMID- 12725071 TI - [Recent progress in the study of common fusarium mycotoxins on apoptosis of cells]. AB - Fumonisin B1, T-2 toxin, Fusarenon-X and Deoxynivalenol, which are all mycotoxins produced by fusarium, could induce apoptosis of liver cell, kidney cell, gastrointestinal epithelial cell, immunological cells as well as several cell lines of human and animals. The possible mechanisms of apoptosis induction are complicated. Sphingol, p21 gene, protein kinase, CAMP signal system and intracellular Ca2+ level, might be involved in the cellular apoptosis induction by Fusarium mycotoxins. PMID- 12725072 TI - [Study on the toxicity of horseshoe crabs in mice]. AB - In order to study the toxicity of horseshoe crabs(tachypleus tridentatus and carcinoscorpius rotundicauda) in the sea of China, the extracts of tissues from tachypleus tridentatus and carcinoscorpius rotundicauda were injected into the abdominal cavity of mice for testing their poisoning effects. The results showed that the toxicity of carcinoscorpius rotundicauda was much higher than that of tachypleus tridentatus. The length of time from the injection to the death was much shorter for Carcinoscorpius rotundicauda than that for tachypleus tridentatus. The signs before death for Carcinoscorpius rotundicauda poisoning were restless, jumping and spasm but that for Tachypleus tridentatus was lethargy. The toxicity of adult horseshoe crabs was much higher than that of young horseshoe crabs. PMID- 12725074 TI - [Assessment on acute toxicity of combined pesticides]. AB - The acute oral toxicity (LD50) of two pesticides combined from 3 categories of insecticides, i.e. organophosphates(OPs), pyrethroids and carbamates, was evaluated by Harris method on equal toxicity doses. The OPs compounds studied included: methylparathion, omethoate, methamidophos, phoxim, dichlorvos, profenofos, isocarbophos and malathion; the pyrethroids: alpha-cypermethrin, deltamethrin, fenvalerate and fenpropathrin; the carbamates: methomyl, isoprocarb and metolcarb. The mixtures of two OPs on the combination of dichlorvos plus omethoate and methamidophos plus profenofos showed additive effects, but methylparathion plus phoxim showed antagonistic effect. Most of the combination of OPs with pyrethroids showed synergistic effects, such as the mixtures of phoxim plus deltamethrin, phoxim plus alpha-cypermethrin, methlyparathion plus alpha-cypermethrin and ioscoarbphos plus fenpropthrin, with the exception of two mixtures showed additive effects: phoxim plus fenvalerate resulting in 1.5 times higher toxicity than expected, while dichlorvos plus deltamethrin resulting in less toxicity than expected. The effects of the mixtures of OPs and carbamates: phoxim plus methomyl and methamidophos plus metolcarb, were all additive, but the mixtures of he methylparathion plus methomyl showed antagonistic effect and malathion plus isoprocarb were synergistic. The biochemical mechanisms of changed toxicity of pesticides mixtures were discussed based on their toxicokinetics and toxicodynamics. It was concluded that the combined effects of insecticide mixtures were additive for OPs plus OPs or plus carbamates in most cased, but synergistic for OPs plus pyrethroids. PMID- 12725073 TI - [Lycopene on cancer prevention]. AB - Many epidemiological research indicate that there is an inverse correlation between the consumption of tomato products and the risk of cancer, especially the gastrointestinal cancer and prostate cancer. The biological features of lycopene, such as molecular structure, metabolism, tissue distribution and its anti-cancer function and possible mechanism are reviewed. PMID- 12725075 TI - [Synthesis and identification of methylparathion artificial antigen]. AB - In order to synthesize the artificial antigen methylparathion(M1605), methylparathion was reduced into amino-methylparathion by using acetic acid-zinc powder-hydrochloric acid. Artificial antigens (M1605-BSA, M1605-TTH) were synthesized by conjugating amino-methylparathion to bovine serum albumin(BSA) and tachypleus tridentatus hemocyanin (TTH) directly after diazotization. Rabbits were immunized with M1605-BSA for 10 weeks, and the high titer and high specificity antiserum from those rabbits was testified by double agar gel diffusion and indirect ELISA. The results showed that an artificial antigen was obtained successfully and this made it possible to establish the immunoassay of M1605. PMID- 12725077 TI - [Effects of glycidyl methacrylate on gap junctional intercellular communication]. AB - In order to probe into the mechanism of glycidyl methacrylate (GMA)-induced cell transformation, the effects of GMA on gap junctional intercellular communication (GJIC) in human lung, embryonic fibroblasts(HLEF) were studied by using scrape loading and dye transfer(SLDT) technique. HLEF cells were treated with GMA at concentrations of 0.5, 2.5 and 5.0 mg/L for 12 hours and Lucifer Yellow was introduced by scrape-loading after exposure. The GJIC in exposed cells and that of control were determined directly as the diffusion extent of fluorescent dye into adjacent cells. The results indicated that GMA could inhibit the GJIC in exposed cells in a dose-dependent manner. Significantly decreased GJIC was found in 2.5 and 5.0 mg/L dose groups. It was suggested that the inhibition of GJIC might be one of the mechanisms responsible for GMA-induced cell transformation. PMID- 12725076 TI - [Changes of pulmonary surfactant in the rat lung exposed to terephthalic acid]. AB - The contents of phospholipids in the lung surfactant in bronchoalveolar lavage fluid(BALF) of rats inhaled terephthalic acid (TPA) dust were analysed with high pressure liquid chromatography (HPLC). The average geometric concentrations of TPA dust in experimental groups were 8.93, 274.45 and 618.12 mg/m3 respectively. The results showed that phosphatidylcholine(PC), phosphatidyl glycerol(PG), and phosphatidylinositol(PI) of the rats exposed to TPA at the middle and high concentrations were decreased as compared with those in the control (P < 0.001). It was suggested that the functions of producing and secreting pulmonary surfactant in type II epithelial cells were damaged after inhalating TPA dust. PMID- 12725078 TI - [Effect of mercury chloride on the reproductive function and visceral organ of female mouse]. AB - In this paper the effects of mercury chloride on the coefficient of visceral organ of mouse and the meiotic maturation and the fertilization ability of mouse oocyte were studied by in vitro culture and in vitro fertilization (IVF) of mouse oocytes, etc. The results showed that mercury chloride (0.5 and 1.5 mg/kg BW) had obviously toxicity to the liver and the kidney of mouse, and the dose of 1.5 mg/kg BW could damage the ovary function and reduce the number of superovulation oocytes. The results also showed that mercury chloride could inhibit the extruding of the first polar body and affect the quality and the viability of mouse oocyte and reduce the rate of IVF, but little impact on germinal vesicle breakdown (GVBD) of mouse oocyte in vivo. The results indicated that mercury chloride could affect the meiotic maturation of mouse oocyte, obviously block the IVF and injury or make a reduction of reproductive capacity of mouse. PMID- 12725080 TI - [The protective effects of zinc metallothionein against erythrocyte membrane damage induced by methylmercury]. AB - The influences of zinc metallothionein(ZnMT) induced by Zn on mice erythrocyte membrane configuration, membrane sulfhydryl content, membrane fluidity, membrane permeability and so on after methylmercury (MeHg) treatment were studied, the protection of ZnMT against the membrane damage of MeHg was studied. The results showed that Zn could induce the production of ZnMT in the liver of mice. With the increase of Zn content, the level of ZnMT increased and had dose-effect relationship between them (r = 0.996). ZnMT could protect membrane configuration and -SH and prevent the damage of MeHg on the membrane fluidity and permeability. The results also showed that the dose of Zn administered to mice should be appropriate. High doses of Zn could cause damages. PMID- 12725079 TI - [Protection of metallothionein on the peroxidative injury in testis induced by cadmium]. AB - In order to study the protection effect of metallothionein (MT) on peroxidative injury in testis induced by cadmium and to detect the mechanism of reproduction toxicity of cadmium, the SD rats were injected intraperitoneally with different doses of cadmium (0.2, 0.4 and 0.8 mg/kg BW) for 7 days. The content of MT, the activities of antioxidases and the content of malondialdehyde(MDA) in testis were detected. The results showed that the increase of MT content in the low dose group was not obvious. MT content increased significantly in the 0.4 mg/kg BW group, while decreased in the 0.8 mg/kg BW group. There was a significant reduction of the activities of superoxide dismutase(SOD) and glutathione peroxidase (GSH-Px) in the high dose group. The content of MDA did not change obviously in the low and middle dose groups, but increased significantly in the high dose group. It was concluded that MT might play an important role in protecting testis from oxidative damage. The toxicity of cadmium on male reproduction might be related to the unbalance of anti-oxidation system including MT. PMID- 12725081 TI - [Kinetics of plasma membrane and mitochondrial alterations in HL-60 cells undergoing apoptosis]. AB - In the present study, VP16, an inhibitor of topoisomerase II, and NaN3, a chemical toxic substance, were used to induce apoptosis and necrosis of HL-60 cells, respectively. Cells were examined by transmission electron microscopy and by fluorescence microscopy after staining with Hoechst 33258 at 0, 2, 4, 8, 16 and 24 h of culture. Alterations of fluorescent intensity produced by FDA, Rh 123 and PI were measured by flow cytometry, which reflects sequential changes on plasma membrane permeability and the potential of mitochondrial membrane. The results indicated that the nuclear morphologic alteration of cells treated with VP-16 began at 4 hour of culture and the cells with condensed nucleus culminated at 8 h of culture and then reduced with no drop in cell counts, while the percentage of cells with fragmented nuclei reached its maximum at 24 h of culture accounting for some 80%. The potential of mitochondrial membrane of of apoptotic cells decreased gradually from 8 h of culture and showed obvious decline at 16 h of culture. In necrotic cells, the potential decreased by 50% at 4 h of culture which indicates an earlier and more rapid decline than that in apoptotic cells. Alteration of plasma membrane permeability appeared at 8 h of culture and showed a steady increase over time. It was concluded that plasma membrane permeability and mitochondrial membrane potential could reflect the development and degree of apoptosis, and the combination of these two criteria with nuclear morphology revealed by staining with Hoechst 33258 would be a simple and reliable assay for apoptosis and its progression. PMID- 12725082 TI - [Effects of pre-heating on the sensitivity of K562 cell to tumor necrosis factor alpha]. AB - In order to study the effects of pre-heating on the sensitivity of cells to tumor necrosis factor-alpha (TNF-alpha), the K562 cells were pre-heated at 40 degrees C for different times to induce heat shock protein 70 (HSP70). To observe the effects of TNF-alpha concentration on the sensitivity of pre-heated cells, cells were pre-heated for 60 min and treated with TNF-alpha (50, 100, 200 and 400 U/ml) for 16 hours, and the cells vitalities were detected by MTT. In order to observe the effect of pre-heating time on the sensitivity of cells to TNF-alpha, the pre heated cells were treated with 100 U/ml TNF-alpha for 24 hours. The levels of HSP70 were tested by reversed transcription-PCR(RT-PCR). The results showed that HSP70 could be induced by pre-heating, and reached its top level after 120 mins, and the sensitivity of non-heated cells to TNF-alpha was lower than that of pre heated cells. To 100 U/ml TNF-alpha, the sensitivity was lowest after the cells were pre-heated for 120 min. The results suggested that pre-heated could induce the expression of HSP70 and inhibit the sensitivity of cell to TNF-alpha, the mechanism may be involved with the overexpression of HSP70. PMID- 12725083 TI - [Effect of hyperthermia on the red-cell immune function of rats and its teratogenicity on developing embryos]. AB - After rats on gestation day 10 exposed to hyperthermia(40 degrees C) for 0.5, 1.0, 2.5 and 5.0 h, red-cell C3b receptor activity(aggregate), red-cell immune complex, circulating immune complex, Nile blue sulfate (NBS) vital staining, electronic microscope and histochemical techniques were used in investigate whether the hyperthermia was disturb red-cell immune functions of pregnant rats and played some role in teratogenesis. The results showed that hyperthermia increased the level of red-cell C3b receptor activity, red-cell immune complex and circulating immune complex. On the other hand, many of them also decreased to normal level after rats were placed in room temperature for 20 h. Following the extension of hyperthermic exposure, the teratogenic incidence increased from 0.9%(1/117) to 18.3%(22/120), in apparent time-effect relationship, when pregnant rats of gestation day 11.5 were killed. It was found that hyperthermia induced. PMID- 12725084 TI - [Synergic effects on primary hepatocellular cancer between history of hepatitis B, family histories of cancers and psychosocial factors]. AB - To study the risk factors of primary hepatocellular cancer (PHC) and their interactions, a case-control study with calculated odds ratios(OR) and synergy effects index(SI) was conducted. The history of hepatitis B, family histories of cancers and psychosocial factors were main risk factors of PHC, and their effects were positive and synergic. The SI between hepatitis B and family histories of cancers, psychosocial factors are 1.65 and 1.16 respectively. PMID- 12725085 TI - [Effects of tea polyphenols on cell kinetics and apoptosis of primary cultured rat skin keratinocytes]. AB - In order to study the effects of tea polyphenols on cell growth and apoptosis of primary cultured rat skin keratinocytes, the safe dosage of tea polyphenols was determined by the release of lactate dehydrogenase(LDH). Cell counting(cytometry) and flow cytometry(FCM) were used to estimate the cell growth and the percentages of cells in different cell cycle phases or apoptosis. The results showed that tea polyphenols stimulated the cell growth from G1/G0 to G2/M phase, enhanced the synthesis of DNA(increase S phase) and increased the proliferative index(PI) from 18.17% to 25.62%. Also, it could block the occurrence of apoptosis. It is concluded that tea polyphenols could stimulate the growth of skin keratinocytes through stimulating the synthesis of cell DNA and inhibiting apoptosis. PMID- 12725086 TI - [Effect of sepia on intracellular Ca2+ concentration, nuclear Ca2+/Mg(2+)-ATPase activities and c-jun expression in H22 cancer cells]. AB - The effect of sepia on the intracellular Ca2+ concentration, nuclear Ca2+/Mg(2+) ATPase activities and the expression of c-jun were studied in H22 cancer cells by using fluorescent probe and immunohistochemical method. The results showed that intracellular Ca2+ concentration decreased 69% and 79%, nuclear Ca2+/Mg(2+) ATPase activities diminished 21% and 37%, c-jun expression decreased obviously. The results suggested that sepia probably diminished the intracellular Ca2+ concentration, affected Ca(2+)-dependent nuclear Ca2+/Mg(2+)-ATPase activities, thus reduced the amount of Ca2+ transporting into nuclei, so lessened the promoting effect of Ca2+ on c-jun expression, and therefore inhibited cellular differentiation and proliferation. This might be one of the possible anti-cancer mechanisms of sepia. PMID- 12725087 TI - [Study on the absorption of environmental contaminants in low-level exposure by pharmacokinetic analysis]. AB - A dynamic generating toxic gas system and a nose-only exposure system were used for the pharmacokinetic study of inhaled environmental contaminants for benzene, toluene, xylene, ethylbenzene, chlorobenzene, styrene, isopropyl benzene, tetrachloroethylene, nonane and methylcyclohexane in male guinea pig. The change of these substances in blood with time was determined simultaneously by solid phase micro-extraction(SPME) gas chromatography (GC). The results showed that the fraction of absorption of benzene at low (121 micrograms/m3) exposure was 4.8 times higher than that at high(12.1 mg/m3) exposure. The pharmacokinetics of these substances were evaluated by using linear compartment models. The data showed that more styrene was absorbed than tetrachloroethylene at low-exposure. The metabolic elimination of these compounds at various exposure concentrations was extrapolated by using estimated pharmacokinetic parameters. Moreover, not only should the differences in absorption quantities be considered in evaluation of potential risk assessment, the metabolic elimination rates should also be considered although the exposure concentrations in gas for all chemicals were equal. The data presented in this paper was fundamental data used for risk assessment. PMID- 12725088 TI - [Effects of electromagnetic radiation from handsets of cellular telephone on neurobehavioral function]. AB - In order to study the effects of electromagnetic radiation from handsets of cellular telephone on neurobehavioral function, 81 staff with handsets of cellular telephone and 63 staff without handsets of cellular telephone from corporations were selected as the subjects. The subjects were investigated by questionnaire on their general health, lifestyle habit, suppress of spirit, handset using of cellular telephone, environmental exposure, morbidity, and the neurobehavioral core test battery(NCTB). The data was analyzed by chi-square, stepwise regression analysis and covariance statistics. The results showed that the average reaction time in user's group was longer than that in control group (P < 0.01). The time of using handset was negatively associated with corrected reaction number (P < 0.01). The fast reaction time and the slowest reaction time were positively associated with the length of handset using (P < 0.01, P < 0.05). The results suggested that the handset using could cause adverse health effects in neurobehavioral function. PMID- 12725089 TI - [Study on plasma lipids, apolipoprotein A1, apolipoprotein B100 in simple obese children]. AB - To investigate the disorder of lipid metabolism and explore the possible mechanism of increasing risk of atherosclerosis(AS) in simple childhood obesity, serum lipids and apolipoprotein were examined. 49 obese children were selected and another 49 non-obese children were selected as control. Their serum lipids (TG, TC, VLDL-ch, HDL-ch and LDL-ch) and apolipoprotein(apoA1 and apoB100) were measured. The results showed TG, VLDL-ch, apoB100, artherosclerosis index (AI) and LDL-ch/HDL-ch ratio in childhood obesity were significantly higher than that in normal children (P < 0.05), while HDL-ch, apoA1, HDL-ch/TC, apoA1/apoB100 ratio in obese children were obviously lower than that in normal ones(P < 0.05). The results suggested that metabolic disorder of lipid and apolipoprotein took place in obese children. The disorder which may increase the risk of AS should be paid great attention. PMID- 12725090 TI - [Effects of zinc deficiency on the c-fos gene expression and transcription in the epiphyses of fetal mouse long bone in culture]. AB - The effects of zinc on c-fos gene expression and transcription in the epiphyses of fetal long bone were studied in mice. The long bones of 16-day fetal mouse were cultured for 48 hours(in medium GBJb) and then used for measuring c-fos gene expression and transcription by immunohistochemistry and in situ hybridization. The result was analyzed by an imaged-analyses system. The experiment was divided into zinc control group(ZC), zinc deficiency group(ZD), zinc deficiency replenish group(ZDR) and zinc stimulatory group(ZS) respectively. The result showed that 1) zinc deficiency caused c-fos protein and mRNA expression and the number of reactive cell decreased. 2) when zinc concentration of medium was 100 mumol/L, the expression and transcription of c-fos gene were increased in hypertrophic, proliferative and resting zone of epiphyses. The research suggested that zinc could affect the expression of c-fos gene in mouse fetal long bone. PMID- 12725092 TI - [Functional aging and compensation of elder workers and ergonomic design]. AB - It was probably inappropriate to use chronological age as the only index for assessing the functional capacities of elder workers. In fact, some functions of elder workers were remained or improved for compensation. The characteristics of functional aging of elder workers were summed up and analyzed. The compensatory mechanisms of accommodation and strategy change, experience and expertise, environmental and social support, and skill learning and retraining were discussed. Finally, the perspective of elder worker ergonomics was depicted in the paper. It is believed that functional age, rather than chronological age, is a scientific index in evaluating the working capacities of elder workers. PMID- 12725091 TI - [Biomarkers in the studies on chemoprevention of colorectal cancer]. AB - The present study was to investigate the chemopreventive effects of tea on colorectal cancer with a series of biomarkers. The results showed that the number of silver-stained nucleolar organizer dots per nucleus(AgNORs), labeling index(LI) of proliferating cell nuclear antigen(PCNA) of intestinal mucosa, and the expression of ras-p21 protein were significantly reduced in the tea-treated groups(P < 0.01) as compared with the positive control group. Furthermore, tea and tea pigments inhibited the expression of Bcl-2 protein and induced the expression of Bax protein(P < 0.05 or P < 0.01). The study provided evidence supporting that PCNA-LI, AgNORs, Bcl-2, Bax and ras-p21 protein could be used as effective biomarkers for colorectal carcinogenesis in human chemopreventive trials. PMID- 12725094 TI - [Spectrophotometric determination of trace nickel in environmental samples by p acetyl arsenazo]. AB - A new spectrophotometric method for the determination of micro nickel p-acetyl arsenazo was studied. The maximum absorption of the Ni-p-acetyl-arsenazo complex was observed at 640 nm with the molar absorptivity of 6.03 x 10(4) L.ml-1.cm-1. The nickel content within 0-0.8 mg/L followed the Beer's Law. PMID- 12725093 TI - [Isolation and purification of Lycium barbarum polysaccharides and its antifatigue effect]. AB - A purified component of lycium barbarum polysaccharide (LBP-X) was isolated from lycium barbarum L. by DEAE ion-exchange cellulose and sephacryl gel chromatography. LBP-X was tested on five different doses (5, 10, 20, 50 and 100 mg.kg-1.d-1) in mice. The results showed that LBP-X induced a remarkable adaptability to exercise load, enhanced resistance and accelerated elimination of fatigue. LBP-X could enhance the storage of muscle and liver glycogen, increase the activity of LDH before and after swimming, decrease the increase of blood urea nitrogen (BUN) after strenuous exercise, and accelerate the clearance of BUN after exercise. The dosage of LBP-X 10 mg.kg-1.d-1 was the best amount among the five tested doses. PMID- 12725095 TI - [Effect of irradiation on the shelf-life of aquatic products]. AB - The effect of irradiation at doses of 0-10 kGy on the shelf-life of aquatic products was determined. The results showed that irradiation could not prolong the shelf-life of aquatic products kept at 25 degrees C. While the aquatic products were kept at refrigerator (4 degrees C), the irradiation could prolong the shelf-life and effectively reduce the total aerobic plate count. Irradiation could effectively reduce the number of coliform to less than 300/kg and eliminate Shigella shigae, Salmonella and Vibrio parahaemolyticus inoculated on shrimp, needle fish and crucian. A dose of 2.5 kGy could delay the rising of total volatile basal nitrogen in samples. PMID- 12725096 TI - [Determination of effective components in propolis]. AB - Propolis is a resinous hive product collected by bees. Propolis is a complex mixture of phenols, acids, alcohol and flavonoids. The aim of the present work is to study the effective components present in propolis and to confirm its biological activities. Four extracts from propolis were obtained by using different organic solutions. Further separation and identification were carried out by capillary gas chromatography and mass spectrometry(CGC/MS). The CGC/MS analysis on propolis extracted by different solvents resulted in more than 80 compounds being separated. Another 38 bioactive materials, including 6 alcohol, 5 phenols, 6 acids, 6 flavonoids and 13 others, have been identified. PMID- 12725097 TI - [A fitting power curve equation for the accumulative inhaling volume in Chinese under 19 years old]. AB - A fitting power curve equation based on the breath frequency and body weight for the accumulative inhaling volume in Chinese under 19 years old was established. The equation is Y = 754.37 + 258.34 X1.9038, and the fitting is good (R2 = 0.9974). It is useful for estimating the degree of exposure to air pollutants of people in their young period. PMID- 12725098 TI - Panoramic radiograph in pathology. PMID- 12725099 TI - Neuroimaging of neck pathology. PMID- 12725100 TI - Angiographic evaluation of the head and neck. PMID- 12725101 TI - Magnetic resonance imaging of the head and neck. PMID- 12725102 TI - Erasmus Darwin, Herbert Spencer, and the origins of the evolutionary worldview in British provincial scientific culture, 1770-1850. AB - The significance of Herbert Spencer's evolutionary philosophy has been generally recognized for over a century, as the familiarity of his phrase "survival of the fittest" indicates, yet accounts of the origins of his system still tend to follow too closely his own description, written many decades later. This essay argues that Spencer's own interpretation of his intellectual development gives an inadequate impression of the debt he owed to provincial scientific culture and its institutions. Most important, it shows that his evolutionism was originally stimulated by his association with the Derby philosophical community, for it was through this group--of which his father, who also appears to have espoused a deistic evolutionary theory, was a member--that he was first exposed to progressive Englightenment social and educational philosophies and to the evolutionary worldview of Erasmus Darwin, the first president of the Derby Philosophical Society. Darwin's scheme was the first to incorporate biological evolution, associationist psychology, evolutionary geology, and cosmological developmentalism. Spencer's own implicit denials of the link with Darwin are shown to be implausible in the face of Darwin's continuing influence on the Derby savants, the product of insecurity in his later years when he feared for his reputation as Lamarckism became increasingly untenable. PMID- 12725103 TI - Rethinking Big Science. Modest, mezzo, grand science and the development of the Bevalac, 1971-1993. AB - Historians of science have tended to focus exclusively on scale in investigations of largescale research, perhaps because it has been easy to assume that comprehending a phenomenon dubbed "Big Science" hinges on an understanding of bigness. A close look at Lawrence Berkeley Laboratory's Bevalac, a medium-scale "mezzo science" project formed by uniting two preexisting machines--the modest SuperHILAC and the grand Bevatron--shows what can be gained by overcoming this preoccupation with bigness. The Bevalac story reveals how interconnections, connections, and disconnections ultimately led to the development of a new kind of science that transformed the landscape of large-scale research in the United States. Important lessons in historiography also emerge: the value of framing discussions in terms of networks, the necessity of constantly expanding and refining methodology, and the importance of avoiding the rhetoric of participants and instead finding words to tell our own stories. PMID- 12725104 TI - "An expedition to heal the wounds of war". The 1919 eclipse and Eddington as Quaker adventurer. AB - The 1919 eclipse expedition's confirmation of general relativity is often celebrated as a triumph of scientific internationalism. However, British scientific opinion during World War I leaned toward the permanent severance of intellectual ties with Germany. That the expedition came to be remembered as a progressive moment of internationalism was largely the result of the efforts of A. S. Eddington. A devout Quaker, Eddington imported into the scientific community the strategies being used by his coreligionists in the national dialogue: humanize the enemy through personal contact and dramatic projects that highlight the value of peace and cooperation. The essay also addresses the common misconception that Eddington's sympathy for Einstein led him intentionally to misinterpret the expedition's results. The evidence gives no reason to think that Eddington or his coworkers were anything but rigorous. Eddington's pacifism is reflected not in manipulated data but in the meaning of the expedition and the way it entered the collective memory as a celebration of international cooperation in the wake of war. PMID- 12725105 TI - Portrait of a discovery. Watson, Crick, and the double helix. AB - This essay examines an iconic image of twentieth-century science: Antony Barrington Brown's photograph of James Watson, Francis Crick, and the double helical model of DNA. The detailed reconstruction of the production, reception, and uses of the photograph reveals the central role of the image in making the discovery it portrays. Taken in May 1953, two full months after the scientists built the model, to accompany a report on the structure in Time magazine, the photograph (like the report) was never published. It came into circulation only fifteen years later, as an illustration in Watson's best-selling book The Double Helix. While the image served as a historical document and advertisement for the book, only the book provided the description that made the image as well as the people and the model it represented famous. The history of the image provides insights into the retrospective construction of the discovery, which has since been celebrated as the origin of a new science of life. PMID- 12725106 TI - "Mom. My tooth!". PMID- 12725107 TI - Alteration in taste: a common oral disorder. PMID- 12725108 TI - Texas dental establishments in 2000. PMID- 12725109 TI - Anxious (patient) about amalgam. Response to ethical dilemma #43. PMID- 12725110 TI - Oral and maxillofacial pathology case of the month. Central ossifying fibroma. PMID- 12725111 TI - Epidemiological investigation of prostate cancer. PMID- 12725112 TI - Human prostate cancer cell lines. PMID- 12725113 TI - Growth of prostatic epithelial and stromal cells in vitro. PMID- 12725114 TI - Prostate epithelial stem cell isolation and culture. PMID- 12725115 TI - Generation of immortal human prostate cell lines for the study of prostate cancer. PMID- 12725116 TI - Spheroids of prostate tumor cell lines. PMID- 12725117 TI - Animal models of prostate cancer. PMID- 12725118 TI - Transgenic mouse models for prostate cancer. Identification of an androgen dependent promoter and creation and characterization of the long probasin promoter-Large T antigen (LPB-Tag) model. PMID- 12725119 TI - In vivo models of human prostate cancer bone metastasis. PMID- 12725120 TI - Effects of fixation on tissues. PMID- 12725121 TI - Background, methods, and protocols for the histopathological diagnosis of prostate carcinoma. PMID- 12725122 TI - Realizing the potential of ejaculate/seminal fluid in detecting and predicting natural history. PMID- 12725123 TI - Bisulfite methylation analysis of tumor suppressor genes in prostate cancer from fresh and archival tissue samples. PMID- 12725125 TI - The androgen receptor CAG repeat and prostate cancer risk. PMID- 12725124 TI - Production and characterization of antipeptide kallikrein 4 antibodies. Use of computer modeling to design peptides specific to Kallikrein 4. PMID- 12725126 TI - Studies on androgen receptor mutations and amplification in human prostate cancer. PMID- 12725127 TI - Proteomics in the analysis of prostate cancer. PMID- 12725128 TI - Application of gene microarrays in the study of prostate cancer. PMID- 12725129 TI - Enhancer trap method using a green fluorescent protein reporter plasmid for cloning tissue-specific enhancers active in prostate cells. PMID- 12725130 TI - Targeted alpha therapy of prostate cancer. PMID- 12725131 TI - Phenotypic and functional differences of dendritic cells generated under different in vitro conditions. PMID- 12725132 TI - Flavonoid compounds in the prevention and treatment of prostate cancer. PMID- 12725133 TI - Position statement: immunomodulator therapy for inflammatory Bowel disease. PMID- 12725134 TI - Percutaneous cholecystostomy in the management of acute cholecystitis. AB - BACKGROUND: The mainstay of therapy for acute cholecystitis is cholecystectomy, which has a mortality of 14-30% in high risk patients. An alternative approach in patients suffering from acute cholecystitis with contraindications to emergency surgery is percutaneous cholecystostomy. OBJECTIVE: To evaluate the efficacy and safety of percutaneous cholecystostomy as the initial treatment of acute cholecystitis in high risk patients. METHODS: Eighty consecutive patients (42 men, 38 women) underwent ultrasound-guided percutaneous cholecystostomy over a 5 year period. Sixty-five patients suffered from acute calculous cholecystitis, 4 patients had acalculous cholecystitis, and 11 patients had sepsis of unknown origin. RESULTS: Sixty-eight patients improved after the percutaneous gallbladder drainage, 10 patients died from co-morbid disease and 2 patients died from biliary peritonitis. During a 1 year follow-up, 32 of the patients underwent interval cholecystectomy, 4 additional patients died from a co-morbid disease, 18 patients did not suffer from any gallbladder symptoms, and 14 were lost to follow up. CONCLUSIONS: Percutaneous cholecystostomy is an effective contribution to the treatment of acute cholecystitis in high risk patients. PMID- 12725135 TI - How much of a misnomer is "asymptomatic" intestinal malrotation? AB - BACKGROUND: Intestinal malrotation is usually observed in the neonatal period with signs of acute high intestinal obstruction due to midgut volvulus. However, malrotation presenting beyond the neonatal period and well into adult life is associated with a variety of atypical and frequently non-specific gastrointestinal symptoms that may often cause prolonged delay in diagnosis and appropriate treatment. OBJECTIVES: To emphasize the difficulty in predicting the risk of midgut volvulus based on age or symptoms, and to recommend surgery in all patients found to have intestinal malrotation even if they are considered asymptomatic. METHODS: We reviewed 41 patients with malrotation treated over a period of 24 years at the Soroka University Medical Center. RESULTS: In our series, 27 patients (66%) had acute midgut volvulus while 14 (34%) had malrotation found during investigation of various long-term gastrointestinal non specific symptoms. Two patients died of total parenteral nutrition-related sepsis following extensive resection of small bowel. A total of 28 patients was available for long-term follow-up and are asymptomatic. CONCLUSIONS: We recommend elective laparotomy and Ladd procedure in all patients found to have intestinal malrotation. This will prevent the catastrophic results of midgut volvulus and a variety of gastrointestinal symptoms wrongly attributed to other conditions in the span of a lifetime. PMID- 12725136 TI - Free perforation in Crohn's disease. AB - BACKGROUND: Free bowel perforation is one of the indications for emergency surgery in Crohn's disease. It is generally accepted that 1-3% of patients with Crohn's disease will present with a free perforation initially or eventually in their disease course. OBJECTIVES: To evaluate the incidence and treatment results of free perforation in patients with Crohn's disease and, based on our experience, to suggest recommendations. METHODS: Between 1987 and 1996, 160 patients with Crohn's disease were treated in our department and were followed for a mean period of 5 years. RESULTS: Of the 83 patients (52%) requiring surgical intervention, 13 (15.6%) were operated due to free perforation. The mean age of the perforated CD was 33 +/- 12 years and the mean duration of symptoms prior to surgery was 6 years. The location of the free perforation was the terminal ileum in 10 patients, the mid-ileum in 2 patients, and the left colon in 1 patient. Surgical treatment included 10 ileocecectomies, 2 segmental resections of small bowel, and resection of left colon with transverse colostomy and mucus fistula in one patient. There was no operative mortality. Postoperative hospital stay was 21 +/- 12 days (range 8-55 days). All patients were followed for 10-120 months (mean 58.0 +/- 36.7). Six patients (42%) required a second operation during the follow-up period. CONCLUSION: The incidence of free perforation in Crohn's disease in our experience was 15.6%. We raise the question whether surgery should be offered earlier to Crohn's disease patients in order to lower the incidence of free perforation. PMID- 12725137 TI - Divergent effects of nicotine administration on cytokine levels in rat small bowel mucosa, colonic mucosa, and blood. AB - BACKGROUND: Chronic nicotine administration has a dual effect on inflammatory bowel disease: augmentation of jejunitis and amelioration of colitis. We previously showed that chronic nicotine administration has divergent regional effects on small bowel and colonic mucosal mediators and blood flow. OBJECTIVE: To examine the effects of nicotine administration on cytokine levels in normal rat small bowel mucosa, colonic mucosa, and blood. METHODS: Male Sprague-Dawley rats weighing 200-250 g were given nicotine (12.5 micrograms/ml) that was dissolved in tap water. Rats were sacrificed on days 1, 2, 7 and 14 after nicotine initiation; blood was withdrawn, and small bowel and colon were resected, washed and weighed. Mucosal scrapings were extracted in 2 ml Krebs Hemselest buffer for determination of interleukins-2, 6 and 10 using the Biosource International Immunoassay Kit. RESULTS: Nicotine decreased IL-10 and increased IL-6 levels in small bowel mucosa (from 3.5 +/- 0.5 to 0.4 +/- 0.1 pg/ml and from 1.9 +/- 0.4 to 13.6 +/- 0.4 pg/ml respectively; P < 0.05). Nicotine decreased IL-2 levels in the colon (from 15.8 +/- 3.0 to 7.9 +/- 1.0 pg/ml; P < 0.05), having no effect on IL-10 or IL-6 levels. Rats treated with nicotine had lower IL-6 and IL-2 blood levels compared to control rats. CONCLUSIONS: Nicotine has different regional effects on small bowel and colonic cytokine mucosal levels, which might explain some of its opposite effects on small bowel and colonic inflammation. PMID- 12725138 TI - The pathophysiology of irritable bowel syndrome--an update. PMID- 12725139 TI - Peptide growth factors and intestinal adaptation in short bowel syndrome. AB - Intestinal adaptation is the term applied to progressive recovery from intestinal failure following a loss of intestinal length. The regulation of intestinal adaptation is maintained through a complex interaction of many different factors. These include nutrients and other luminal constituents, hormones, and peptide growth factors. The current paper discusses the role of peptide growth factors in intestinal adaptation following massive small bowel resection. This review focuses on the mechanisms of action of peptide growth factors in intestinal cell proliferation, and summarizes the effects of these factors on intestinal regrowth in an animal model of short bowel syndrome. PMID- 12725140 TI - Malignant tumors of the small intestine--new insights into a rare disease. PMID- 12725141 TI - Re-eradication of Helicobacter pylori infection following initial treatment failure: treatment options in clinical practice. PMID- 12725142 TI - Non-steroidal anti-inflammatory drug-induced gastroduodenal toxicity and Helicobacter pylori. PMID- 12725143 TI - Is validation of Helicobacter pylori eradication necessary? PMID- 12725144 TI - Irritable bowel syndrome: epidemiology and diagnosis. PMID- 12725145 TI - Adult celiac disease presenting with intussusception and elevated liver enzymes. PMID- 12725146 TI - Perihepatic lesions mimicking primary liver tumors. PMID- 12725147 TI - Liver abscesses caused by Streptococcus milleri: an uncommon presenting sign of silent colonic cancer. PMID- 12725148 TI - Ischemic hepatitis induced by severe anemia. PMID- 12725149 TI - Right gastroepiploic artery stab wound presenting as massive hemothorax: a diagnostic dilemma. PMID- 12725150 TI - Ethical aspects of percutaneous endoscopic gastrostomy insertion. PMID- 12725151 TI - Patient informed consent for digestive endoscopy. PMID- 12725152 TI - Fibrolamellar hepatocellular carcinoma--an uncommon tumor. PMID- 12725153 TI - Alternative medicine. PMID- 12725154 TI - Unusual manifestations of Hodgkin's disease. PMID- 12725156 TI - [The study on expression of caspase-1 in experimental brain contusion of different severity in rats]. AB - OBJECTIVE: To investigate the expression of caspase-1 in different posttraumatic intervals and severity of brain injury. METHODS: The study examined brain tissue samples of slight (n = 24), severe (n = 24) brain injury and control (n = 6) of rat, using immunohistochemical staining, western-blot and RT-PCR method. RESULTS: Up-regulating of caspase-1 expression was found in tissue from traumatic brain injury compared with controls in early 1 hour after TBI, and lasted for 2 weeks. Caspase-1 cleaved after brain injury. CONCLUSION: These findings are the first time to demonstrate the study of different intense brain injury. The increasing of caspase-1 expression indicates that the TBI exists. The threshold area of caspase-1 positive cells is different in different severity of brain injury. PMID- 12725155 TI - [GDNF expression following the severe brain injury in rats]. AB - OBJECTIVE: To evaluate the changes of GDNF after severe brain injury. METHODS: Changes GDNF in cortex, thalamus and pontine was observed by using immunohistochemistry and image analysis technique. RESULTS: The lever of GDNF was increasing at 1 d, reaching peak at 3 d, not obviously decreasing at 5 d and still higher than normal at 7 d after severe brain injury. CONCLUSION: As an objective indication, the time sequence regularity of GDNF after brain injury may be applied in brain injury time estimation. PMID- 12725157 TI - [A study on the expression of C-FOS protein after experimental rat brain concussion]. AB - OBJECTIVE: To study the relationship between expression of C-FOS protein and brain concussion and find a sensitive marker of diagnosis of the brain concussion. METHODS: Fifty-five rats were randomly divided into brain concussion groups and control group. The expression of C-FOS protein was microscopically observed by immunohistochemical method. RESULTS: There were negative expression of C-FOS protein in control group. In brain concussion group, however, positive expression of C-FOS protein in some neurons was seen at 15 min after brain concussion, and reach to the peak at 6 h after brain concussion, then decreased gradually. CONCLUSION: These findings suggest that detection of C-FOS protein could be an index of diagnosis of brain concussion and a sensitive marker of timing of injury after brain concussion. PMID- 12725158 TI - [A analysis of ELISA on the time-related expression of IL-2 and TNF-alpha during the healing process of wound in rat skin]. AB - OBJECTIVE: In order to explore the relationship between the expression change of cytokines and the wound age during the healing process of rats skin wound. METHODS: A assay of ELISA were performed on intravital skin wounds(after incision 0.5-168 h) to detect the dynamics expression level of IL-2, TNF-alpha. RESULTS: The level of IL-2 and TNF-alpha increased at 0.5 h after wounding, then got to a peak at 3 h and 1 h after injure respectively. Rebound of TNF and IL-2 levels were shown at 48 h after wounding, and both cytokine levels were inclined to elevating between 72 h and 168 h after wounding. CONCLUSION: The cytokine level changes suggest they have a time-related expression during wounds healing process. PMID- 12725159 TI - [A study on the expression of VEGF in the blunt injury of rat's skin]. AB - OBJECTIVE: To evaluate changes of VEGF in skin after blunt injury. METHODS: The rats of injury groups were subjected to skin blunt injury by free-falling iron hammer. The samples taken at 1 h, 3 h, 6 h, 12 h, 1 d, 3 d, 5 d, 7 d, 9 d after injury were studied by immunohistochemistry and MIAS image analysis system. RESULTS: In the skin of normal control group the expression level of VEGF was low. The increase of VEGF could be observed at 1 day after injury, reached peak at 7 days and declined at 9 days. CONCLUSION: Blunt injury in the skin could induce the expression of the VEGF. Moreover, the change pattern of VEGF level was quite regular with time and could be used to estimate the time after skin injury. PMID- 12725160 TI - [Age determination by radiographic feature of thyroid cartilage in male Chinese Han popullation]. AB - OBJECTIVE: To seek a exact method of age determination. METHODS: This sample consists of 73 male Chinese han individuals whose ages have been known. Eight positions were selected from their radiograms which have been divided into three or six grade and then set up a mathematical model to deduce age by multiple linear regression analysis. RESULTS: The standard deviation of male thyroid cartilage is 1.854 and the multiple correctation coefficient is 0.9971. CONCLUSION: The observation proved that age determination by radiographic feature of thyroid cartiage may be suitable for male Chinese han population from 13 to 67 years old. PMID- 12725161 TI - [Morphological changes of tissues after extensive soft tissue injury]. AB - OBJECTIVE: To study the pathological morphological changes for diagnosing the cause of death of extensive soft tissue injury or crush syndrome. METHODS: The tissues were stained by HE and IHC. RESULTS: (1) The Mb positive rate was 60%, 75%, 95% respectively. (2) Both the HSP70 positive rate of hearts and brains were 90%. CONCLUSION: (1) The animal model of broad soft tissue injury was established. (2) Accumulated the pathological morphological data for diagnosing the cause of death of extensive soft tissue injury or crush syndrome. PMID- 12725162 TI - [The study of distortion products otoacoustic emissions in pseudo-anacousia]. AB - OBJECTIVE: To explore the technique that distinguish pseudo-anacousia. METHODS: 60 cases were divided into three groups (normal group, deafness sensorineura group and pretending hearing loss group). They all had been tested by distortion products otoacoustic emissions technique. RESULTS: (1) The DPOAE incidence of normal group is 94.00%. the DPOAE incidence of deafness sensorineura group is 22.00%. There were significant differences at statistics in DPOAE incidence (P < 0.01). those proved that patients (deafness sensorineura group) were in hearing problem. (2) The DPOAE incidence of pretending hearing loss group is 93.00%, there were no significant differences at statistics in DPOAE incidence between normal group and pretending hearing loss group (P > 0.01), furthermore when behavior threshold is over 50 dBHL, DPOAE incidence is no score. CONCLUSION: This study shows the technique can distinguish pretending hearing loss. The application of DPOAE can be used as a new tool in cases assessment of clinic forensic medicine. PMID- 12725163 TI - [Polymorphism of PentaD and PentaE STR locus in five Chinese Han population]. AB - OBJECTIVE: To obtain the genetic polymorphism data of Guangxi, Hunan, Henan, Sichuan, Taiwang Chinese Han population and compare the polymorphism of PentaD and PentaE STR locus. METHODS: The two loci was analyzed by using the PowerPlex 16 System. RESULTS: 10 alleles of PentaD and 19 alleles of PentaE were found in the five Han population. PentaD and PentaE have the expected heterozygosity values of 0.7746-0.8047 and 0.9005-0.9219, the polymorphism information content values of 0.7710-0.8025 and 0.8969-0.9176, the discrimination power values of 0.9223-0.9341 and 0.9471-0.9782, the power of exclusion values of 0.5435-0.6325 and 0.6785-0.8465, respectively. CONCLUSION: The result showed that these two loci were highly informative and suitable for forensic application. PMID- 12725164 TI - [New improved approaches for DYS385 detection]. AB - OBJECTIVE: To establish more sensitive methods for detection of DYS385 in routine forensic casework. METHODS: The primers recommended by Genome Database (GDB) and Schneider were used to amplify DYS385 respectively. Then, a semi-nested PCR of DYS385 was designed by using the two different primers as outer and inner primer. A series of experiments were carried out to achieve good result by adjusting the ratio of outer/inner primer and optimizing the PCR condition. RESULTS: It showed that an overall 112 bp shorter DYS385 fragments and better electrophoretic separation were obtained by using primer2B. By using the semi-nested PCR approach, the shorter specific DYS385 fragments could be amplified and detectable DNA amounted to 50 pg. CONCLUSION: This method is 20 fold more sensitive than the ordinary method. PMID- 12725165 TI - [HLA gene polymorphism and forensic medicine]. AB - The gene complex of Human Leukocyte Antigen (HLA) is located of chromosome 6p21, which is the most complicated dominant polymorphic genetic system. The HLA system has 108 genotypes. It is the best human genetic marker. It has been applied to forensic paternity test and individual identification. This article discusses the research development of HLA polymorphism and its application in forensic medicine. PMID- 12725166 TI - [Trauma brain injury and apoptosis]. AB - The evidence and the feature of apoptosis following tyrauma brain injury(TBI) and the possible mechanisms underlying apoptosis were reviewed. Recently research showed that apoptosis play an important role in TBI, the occurring time and area of apoptosis were found significant differences compared with that of necrosis. The neural cell apoptosis can undergo following many pathways after TBI. In our review, the foreground of apoptosis after TBI research in forensic pathology were also discussed. PMID- 12725167 TI - [Traumatic brain injury and caspase]. AB - Many pathologic and physiologic changes occur after brain injury. Many genes control these changes. Caspase plays an important role in neuron cell apoptosis and has concern with secondary brain injury. It is of great significance in the forensic science. PMID- 12725168 TI - [The latest research progress on CGRP and its potential application in forensic medicine]. AB - Calcitonin gene-related peptide (CGRP) play a key role in some physiological and pathological progresses. The latest studies indicate that CGRP might involve in some disease progress and has a close relation with wound healing. It is significant to further investigate and then apply it to clinical diagnosis and therapy as well as forensic pathology. PMID- 12725169 TI - [The sudden cardiac death in negative autopsy]. AB - The advancement of studies about the molecular biology and electronic physiology on sudden cardiac death was summarized in this article, including particularly cardiac concussion(commotio cordis), congenital long QT syndrome, and Brugada syndrome which probably resulting in fatal arrhythmia and sudden cardiac death. These corpses of fatal functional disorders often show the results of negative autopsy without obvious organic pathological changes. So when come across negative autopsy the medical examiner and the pathologist should be careful to investigate the inductive cause of sudden death, the history of disease, and the family history, then to rule out the possibility of the above disorders. PMID- 12725170 TI - Informed consent. Why you need more than a signature. PMID- 12725171 TI - Lasers: more than meets the eye. PMID- 12725172 TI - American dentists reach out to Armenia. PMID- 12725173 TI - Hygienists push to unionize: for 'common good'--or code for independent practice? PMID- 12725174 TI - [Simultaneous posterior and horizontal canal benign paroxysmal positional vertigo]. AB - OBJECTIVE: To explore effective methods for the diagnosis and treatment of simultaneous posterior and horizontal canal benign paroxysmal positional vertigo (combined BPPV, C-BPPV). METHOD: Epley's maneuver and Barbecue rotation were applied to four cases of C-BPPV separately with an interval of one day. RESULTS: Positional vertigo in all subjects disappeared completely after treatment and yielded excellent resolution of symptoms during period of follow-up. CONCLUSION: Clinical features in C-BPPV are combinations of both posterior and horizontal canal BPPV. Combined particle repositioning procedures of Epley's maneuver and Barbecue rotation is a successful method for treating the disorder. PMID- 12725175 TI - [Surgical management of Mondini dysplasia with cerebrospinal fluid leakage]. AB - OBJECTIVE: To evaluate the clinical manifestation and surgical technique of Mondini dysplasia with cerebrospinal fluid (CSF) leakage. METHOD: Three Mondini dysplasia with spontaneous cerebrospinal fluid leakage were treated by the authors. A transcanal tympanotomy were used to pack the vestibular cavity with muscle. RESULT: Three children were first manifested with recurrent meningitis and spontaneous cerebrospinal fluid rhinorrhea. Further examinations found that the leakages were otogenic. One side or both sides Mondini dysplasia were confirmed by CT scanning of temporal bones. The leakages were all stopped by the primary surgical closure. CONCLUSION: Mondini dysplasia should be considered in children with spantaneous cerebrospinal fluid otorhinorrhea. A temporal bone CT scan can confirmed the diagnosis. A transtympanic closure is very effective in the management. PMID- 12725176 TI - [Multi-channel cochlear implantation in patients with common cavity malformation]. AB - OBJECTIVE: To report the multi-channel cochlear implantation in patients with Common cavity malformation, including surgeries, pre and postoperative hearing assessments as well as outcomes of speech recognition. METHOD: Multi-channel cochlear implantation surgeries have been performed in 6 cases with common cavity malformation in our hospital from May 1995 to May 2002. The transmastoid lateral semicircular canal approach was used in these cases. The postoperative outcomes of these cases are compared with 6 cases of non-malformed cochlea as a control group. RESULT: Among the 6 cases, All 32 electrodes were inserted in 3 cases, 28 electrodes in 1 cases, 26 electrodes in 1 cases, 22 electrodes in 1 cases. The intra and postoperative complications of the surgeries have no significant difference compared with the surgeries of normal cochlea. The intraoperative "Ozze" occurred in 2 cases, the small CSF flowing occurred in 3 cases and no CSF flowing occurred in 1 case. Postoperative threshold in sound field were 30-40 dB recovery for all cases, and similar to the normal cochlea cases. Most of them received the similar hearing and speech recovery, compared with the result of non malformed group. Only a few had a higher hearing threshold. The remote effect will be evaluated by long-term follow-up. CONCLUSION: Multi-channel cochlear implantation could be performed in the cases with common cavity malformation, but preoperative hearing assessments and proper images should be done. PMID- 12725177 TI - [Application of computer land mark guide system in oto-rhino-surgery]. AB - OBJECTIVE: To explore related issues of image-guidance system in oto-rhino surgery application through 5 cases of ear and nose diseases. METHOD: All operations were carried out with endoscope or microscope under the guidance of computer aided landmark system. Five patients include one fungus sphenoid sinusitis, one enormous cyst of ethmoid-frontal sinus and three congenital external meatus block. RESULT: Preoperative preparation time is 16-30 minutes, with an average of 27 minutes. The quotient of calibration is 1.3-2.0, with an average of 1.92. Precise error between image and entity are less than 1 mm in base of skull, mastoid; less than 1.3 mm in nasal cavity and sinuses. Operations were successfully finished without complication intraoperatively or postoperatively. The time spent on surgery with image-guidance system is as similar as that of conventional way. CONCLUSION: Image-guidance system is useful for doctors to precisely recognize the mark of high dangerous area in nose, ear and surroundings, which is helpful to improve both the accusation and safety. It is one of the developing directions of microsurgery on otolaryngology field in the future. PMID- 12725178 TI - [Clinical analysis of 48 cases with Mondini dysplasia]. AB - OBJECTIVE: To improve the ability to diagnosis the Mondini dysplasis, 48 cases of Mondini dysplasis were reported. METHOD: The clinical data of 48 cases were collected. The clinical manifestation and the examination of CT and MRI and treatment of the Mondini dysplasia were discussed. RESULT: Most of them were deaf when they were babies. The prolonged speaking and unsteading were important clinical feature. CONCLUSION: The diagnosis of the disease is based on the examination of the hight resolution tomography and magnetic resonance image of the temporal bone. Hearing aid is common management to the disease. A part of patients should be considered cochlear implantation. PMID- 12725179 TI - [Psychological aspects of subjective tinnitus]. AB - OBJECTIVE: To investigate the psychological aspects of tinnitus and so as to treat them more effectively. METHOD: A total of 225 patients with first complain of subjective tinnitus was studied with interviewing, Zuang's self-rating scale of depression and tinnitus questionnaire to evaluate their psychological issues. RESULT: All patients showed obvious psychological problems. 21% of patients were resulted from psychological causes. 67.7% of patients were resulted in server psychological responses. The rest patients showed not only psychological causes but also psychological responses. CONCLUSION: Our results suggest that there is close relationship between tinnitus and psychological issues. Tinnitus questionnaire and tinnitus retraining therapy are powerful tools in diagnosing and treating psychological issues in tinnitus patients. PMID- 12725180 TI - [Study on distortion product otoacoustic emissions and expanded high frequency audiometry in noise exposure workers]. AB - OBJECTIVE: To analyse the relationship of distortion product otoacoustic emissions(DPOAE), conventional pure tone audiometry and expanded high frequency audiometry, and discuss the originated mechanism of DPOAE and value in early diagnosis and detection noise-induced deafness. METHOD: DPOAE, conventional pure tone audiometry and expanded high frequency audiometry were performed in 42 young adults with normal hearing of control group and 20 noise exposure workers of test group. DPOAE amplitudes, conventional pure tone hearing thresholds and expanded high frequency hearing thresholds were compared. RESULT: In noise exposure workers against young adults with normal hearing, the pure tone hearing thresholds at 6.0 kHz and expanded high frequency area declined significantly(P < 0.05), DPOAE amplitudes at frequency from 4.0 to 6.0 kHz and 11.2 kHz declined significantly too(P < 0.05), but there existed no significantly difference at frequency from 12.5 to 16.0 kHz between the two groups(P > 0.05). CONCLUSION: DPOAE is potential implement to early diagnose and detect noise-induced deafness. It seems likely that the origination of DPOAE not limited to the outer hair cell in corresponding cochlear area. The originated area and mechanism of DPOAE should be solved by further studies. PMID- 12725181 TI - [Cholesteatoma in petrous part of temporal bone]. AB - OBJECTIVE: To explore the clinic characteristic of petrous bone cholesteatoma. METHOD: One case was treated by supra-labyrinth approach. 8 cases did by labyrinth-cochlea approach. RESULT: After 2 to 9 years follow-up in the 8 cases, no local cholesteatoma relapsion occurred. 2 cases were treated by facial nerve decompression and facial paralysis did not recover postoperatively. 1 case suffered from facial muscle movement 2 months after operation and recovered after 7 months later. 1 case still needs further follow-up with 2 months postoperatively. CONCLUSION: Proper surgical approach should be selected according to location and invasion of disease. Packing with adipose tissue and blocking the external auditory meatus are effective methods to prevent the cerebrospinal fluid leak. PMID- 12725182 TI - [Measurement of transient evoked otoacoustic emissions and follow-up of 215 healthy newborns]. AB - OBJECTIVE: To evaluate the characteristics of transient evoked otoacoustic emissions (TEOAE) of healthy newborns and improve hearing screening of newborns. METHOD: To investigate the history of pregnancy and parturition, TEOAE was measured in 425 ears of 215 healthy newborns from 0 to 8 days postpartum and follow-up study was undertaken. RESULT: The general prevalence was 89%. There was some relationship between prevalence and testing time. The average prevalence was 74% in newborns of 0-3 days. The prevalence was 97% in newborns of 4-8 days. The way of parturition, laterality and time of pregnancy over 36 weeks hadn't any significant effect on the prevalence rate. The prevalence of females was higher than males. Quick screen was more sensitive than TEOAE. One ear of hearing loss was diagnosed after follow-up. CONCLUSION: The hearing screening test of newborns should be taken in hospitals after 4 days of birth, the way of parturition had no effect on the cochlear function. The mature time of cochlear hair cells was by the 36th week of pregnancy. Quick screen couldn't take place of TEOAE. Someone who had no TEOAE in one ear or two ears must be followed up and reexamined and exact diagnosis could be made as soon as possible combining with ABR. PMID- 12725183 TI - [The endoscopic technique utilized in removal process of acoustic neuroma by retrosigmoid approach]. AB - OBJECTIVE: Research into the technique and signification of reducing remnant tumor with utilizing endoscopic technique during removal of acoustic neuroma. METHOD: 15 patients proceeded the removal of acoustic tumor through retrosigmoid approach (as routine retrosigmoid-approach operation group, RRSO-G), 11 patients utilized endoscope for inspecting and eliminating remain of tumor during the proceed of the removal of acoustic neuroma through retrosigmoid approach (as retrosigmoid-approach operation combined using endoscope, RSOCE-G), all of them were examined of auditory level, vestibular function, facial nerve function, and MRI before and three months after operation. RESULT: The remain tumor were found in 3 cases of RRSO-G and no any one in RSOCE-G after operation. The rates of auditory and vestibular dysfunction, and facial paralysis have no difference between RRSO-G and RSOCE-G. CONCLUSION: It is one of expedient and safe surgery methods that the acoustic neuroma are removed through the retrosigmoid approach, and some degrees of hearing function are reserved at same time. If the endoscope is utilized during this operation, the rate of tumor remaining and neuroma recrudescing could be reduced evidently. The acoustic neuroma removed through the retrosigmoid approach combining with utilizing endoscope is a worthy method for using widely in otoneurosurgery. PMID- 12725184 TI - [Surgical reconstruction of the abnormalities of earlap, canal and middle ear at one time]. AB - OBJECTIVE: To investigate the surgical reconstruction of the abnormalities of earlap, canal and middle ear at one time. METHOD: 21 patients(21 ears) operated on the canal and middle ears, of which 6 ears were undergone surgical reconstruction of the abnormalities of earlap, canal and middle ear used with the Medpor and PTFE at one time. RESULT: A term(8-23 months) follow-up of 6 ears demonstrated that 28 dB of the hearing level in 5 ears were improved (pure tone threshold), one ear was kept unchanged, which 6 ears were approved of the figuration of earlap and canal ears. CONCLUSION: It is an good method to reconstruction the abnormalities of canal, middle ear and earlap using the Medpor and PTFE at one time. PMID- 12725185 TI - [Ototoxic study spiral ganglion neurons in the pig guinea with ethylene oxide remained in absorbable gelatin sponge]. AB - OBJECTIVE: To measure ethylene oxide remained in absorbable gelatin sponge, and to observe change of ultrastructure in spiral ganglion neurons after deposited in otic vesicle of pig guinea with ethylene oxide sterilized absorbable gelatin sponge. METHOD: To use comparative colour technique measuring ethylene oxide remained in absorbable gelatin sponge, by means of transmission electron microscope to observe change of ultrastructure in spiral ganglion neurons after 3 months deposited in otic vesicle of pig guinea with ethylene oxide sterilized absorbable gelatin sponge. RESULT: The mean value of ethylene oxide remained in absorbable gelatin sponge were 438.32 mg, while nation standard are less 10 mg, FDA standard(1978) less 25 mg. ultrastructural appearance showed condensed matric electron dense of spiral ganglion mitochondria and dark crista of mitochondria, vacuolated cytoplasm. Ultrastructural observation also demonstrated dissolution or necrosis of ganglion cells, satellite cells, and vacuolated myelin. These changes were near Donson reported(1997) intracochlear perfusion with aminoglycosides. CONCLUSION: The mean value of ethylene oxide remained in absorbable gelatin sponge were 40 times nation standard. Toxic changes of ethylene oxide remained in absorbable gelatin sponge in spiral ganglion neurons were similar to intracochlear perfusion with aminoglycosides. PMID- 12725186 TI - [Possible role of transforming growth factor alpha on the cholesteatoma growth regulation]. AB - OBJECTIVE: To investigate the mode of expression and distribution of transforming growth factor alpha in middle ear cholesteatoma. METHOD: The specimens from the cholesteatoma tissue of 32 cases and the normal external ear skin of 10 cases were examined by immunohistochemical S-P method and computer image analysis. RESULT: In cholesteatoma epithelium staining for TGF alpha was consistently stronger than that in normal epithelium. The mean absorbance of the two epithelial tissue is 2.431 +/- 0.587 and 1.463 +/- 0.147 respectively, revealing a significant difference. Some immune cells infiltrating the stroma of cholesteatoma also stained positively for TGF alpha. CONCLUSION: These data suggested that an autocrine and paracrine stimulation of the squamous epithelium of cholesteatoma by TGF might contribute to its hyperproliferation. PMID- 12725187 TI - [Trans-external ear canal blow oxygen method]. AB - OBJECTIVE: To study a new method for the treatment of the glue ear and the other out and middle ear diseases. METHOD: 28 ears of the glue ear were treatment with blow oxygen method trans-external ear canal after myringotomy with grommet insertion. The results were compared with that of only tube insertion group of 20 ears in same period. This method were also used to treatment the patients with otitis media after mastoidectomy to compared the effect on dry ear time. RESULT: The therapy time of the blow oxygen group for glue ear were 1.20 +/- 0.42 days, and the control group were 3.50 +/- 0.87 days. The blow oxygen group need only 3.92 +/- 0.77 weeks for dry ear after mastoidectomy. And the control group need 5.82 +/- 2.03 weeks. The results are statistic significant between this two groups. CONCLUSION: The blow oxygen method is a good way for the treatment of glue ear, and can short the time for dry ear after mastoidectomy. PMID- 12725188 TI - [Proteomics analysis and biochemical activity detection on facial nerve regeneration fluid]. AB - OBJECTIVE: To explore the protein components and the biological neutrional activity upon motor neuron, so that take a general, basic study on the nerve regeneration microenvironment which be important to facial nerve regeneration. METHOD: The animal model of rabbit facial regeneration microenvironment is established. Advanced technology of two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) was used to study on the structural proteins in the regeneration fluid, as well as the motor neuron culture was used to test the biological activity in it. RESULT: 850 +/- 78 protein spots were detected in the regeneration fluid. The body area, neurite length and OD value of experimental group are higher than that of control group. CONCLUSION: We got the protein image of facial nerve regeneration fluid by 2-D PAGE, and proved that many kinds and number of proteins in the regeneration fluid have neutrional activity on motor neuron. PMID- 12725189 TI - [Restricted microcirculation disorder induced by photochemical reaction in the guinea pig cochlea]. AB - OBJECTIVE: To investigate the effect of restricted cochlear microcirculation disorder on the cochlear blood flow, auditory function, and morphology. METHOD: Photochemical reaction was utilized to induce localized microcirculation damage to the second cochlear turn. After systemic administration of rose bengal, the second cochlear turn was illuminated with green light (wave length 540 nm +/- 40 nm) to cause thrombosis in the stria vascularis, the area was about 1.2 x 1.0 mm2. RESULT: At the site of the illumination, cochlear blood flow gradually decreased, and the threshold of action potential gradually increased. Surface preparing showed thrombus was in the stria vascularis. Three hours after the photochemical reaction, some hair cells in the illuminated area necrotized. The length of the lesion was 1152.50 +/- 363.26 microns (n = 4). But hair cells in the non-illuminated area were normal. CONCLUSION: The photochemical method is a reliable tool to produce an animal model with the restricted microvessel lesion in the lateral wall of the cochlea. It could be used as an animal model for the treatment of the cochlear thrombosis. PMID- 12725190 TI - [The simultaneous isolation of outer hair cells and Deiters' cells from the cochlea of the guinea pig]. AB - OBJECTIVE: To investigate the simultaneous method of isolating outer hair cells and Deiters' cells from the organ of Corti of the guinea-pig cochlea. METHOD: The organ of Corti was dissected from the pigmental guinea pig cochlea, and then treated with papain. RESULT: A large number of living outer hair cells and Deiters' cells were obtained synchronously from the guinea pig. CONCLUSION: The key to success in isolating outer hair cells and Deiters' cells from the cochlea is to know very well the anatomical characteristics of the cochlea and keep the contact the organ of Corti. PMID- 12725191 TI - Recognizing and rewarding nurse preceptors in critical care. Some answers. PMID- 12725192 TI - Homocysteine testing. PMID- 12725193 TI - Monitoring delirium in critically ill patients. Using the confusion assessment method for the intensive care unit. PMID- 12725194 TI - Teaching documentation tool: building a successful discharge. PMID- 12725195 TI - Effects of patient-controlled music therapy during coronary angiography on procedural pain and anxiety distress syndrome. PMID- 12725196 TI - Psychological influences in critical care: perspectives from psychoneuroimmunology. PMID- 12725197 TI - Advances in cardiac surgery: valve repair. PMID- 12725198 TI - Epilepsy and pregnancy: maternal and fetal effects of phenytoin. PMID- 12725199 TI - Pain management in the critically ill. PMID- 12725200 TI - Examining nursing malpractice: a defense attorney's perspective. PMID- 12725201 TI - Preoperative considerations: which herbal products should be discontinued before surgery? PMID- 12725202 TI - Supporting lifelong learning and competency development. PMID- 12725203 TI - The challenge of urban family medicine. PMID- 12725204 TI - Anatomy of the Medicare appeals process. PMID- 12725205 TI - The integrated summary: a documentation tool to improve patient care. PMID- 12725206 TI - Making family practice doable in everyday life. PMID- 12725207 TI - Evaluating health plans: finding the keepers. PMID- 12725208 TI - Making paper charts more efficient. PMID- 12725209 TI - Keeping the human touch. PMID- 12725211 TI - Second circuit remands malpractice suit arising from ERISA Plan's denial of preauthorization. PMID- 12725210 TI - A saga in international HIV policy modeling: preventing mother-to-child HIV transmission. PMID- 12725212 TI - Community-acquired bacteremia with an ESBL-producing Escherichia coli strain--a case report. PMID- 12725213 TI - Isolation of Salmonella enterica serotype Worthington from empyema fluid. PMID- 12725214 TI - Subcutaneous abscess caused by extended-spectrum beta-lactamase-producing Salmonella senftenberg in a type 2 diabetic--a case report. PMID- 12725215 TI - Isolated cerebrospinal fluid eosinophilia due to Streptococcus bovis meningitis in an infant. PMID- 12725216 TI - Disseminated fungemia due to Candida krusei with cutaneous lesions and successful treatment by amphotericin B lipid complex and catheter removal: a case report. PMID- 12725217 TI - The pattern of transmission transfusion virus infection in Egyptian patients. PMID- 12725220 TI - Shedding light on epilepsy: advocacy groups are pushing to raise awareness about epilepsy and encourage legislatures to make it a more integral part of public health programs. PMID- 12725219 TI - Abnormal Pap test results. PMID- 12725218 TI - Nurses who provide genetics counseling need ongoing education and certification. PMID- 12725221 TI - Safe, but not sound: limiting safe harbor immunity for health and disability insurers and self-insured employers under the Americans with Disabilities Act. PMID- 12725222 TI - Atypical squamous cells-unspecified Pap test and human papillomavirus testing dilemma. PMID- 12725223 TI - Utilizing research to guide clinical practice in prostate cancer education. PMID- 12725224 TI - Prevent Abuse and Neglect through Dental Awareness (P.A.N.D.A.). PMID- 12725225 TI - A tough one to tackle. PMID- 12725226 TI - Piece offering. PMID- 12725228 TI - Editorial. Ethical issues in gastroenterology. PMID- 12725227 TI - Bridge building. PMID- 12725229 TI - Nurses need more education in healthcare policy making. PMID- 12725230 TI - Nitric Oxide in Hypertension and other Cardiovascular Diseases. Prague, June 23, 2002. Abstracts. PMID- 12725231 TI - Proceedings of the 1st International Symposium on Cardiovascular Science--From Bench to Bedside. November 23-24, 2001, Hong Kong. PMID- 12725232 TI - Proceedings of the 21st Annual International Gravitational Physiology Meeting, Nagoya, Japan, April 3-8, 2000. PMID- 12725233 TI - Proceedings of the 16th Meeting of the Working Group for Pharmacology in Oncology and Hematology. Munster, Germany, June 21-22, 2002. PMID- 12725235 TI - Proceedings of the International Symposium on Antimutagenesis and Anticarcinogenesis. April 25-26, 2002, Valhalla, New York, USA. PMID- 12725234 TI - [Umbilical cord blood donation: unrelated donation, familial (directed) donation and autologous donation (private banking)]. PMID- 12725237 TI - Hypoglycaemia 2001: From Research to Practice. Assisi, Italy, 26-29 May 2001. Proceedings. PMID- 12725236 TI - Profiling centres of excellence in CAM research. PMID- 12725238 TI - Claimant identification pilot projects. Final rules. AB - We are revising our regulations that pertain to the processing of initial claims for disability benefits under title II (Social Security Disability Insurance) and title XVI (Supplemental Security Income) of the Social Security Act (the Act). We will be conducting pilot projects wherein we will request photographic identification from individuals filing for title II and title XVI disability and blindness benefits in specified geographic areas covered by the pilot projects. In addition, we will require individuals to allow us to take their photograph and we will make these photographs a part of the claims folder. We will permit an exception to the photograph requirement when an individual has a sincere religious objection. This process will strengthen the integrity of the disability claims process by helping to ensure that the individual filing the application is the same individual examined by the consultative examination (CE) physician. PMID- 12725239 TI - Exemption of chemical mixtures containing the List I chemicals ephedrine, N methylephedrine, N-methylpseudoephedrine, norpseudoephedrine, phenylpropanolamine, and pseudoephedrine. Final rule. AB - On September 16, 1998, the Drug Enforcement Administration (DEA) published a Notice of Proposed Rulemaking (NPRM) to implement provisions of the Controlled Substances Act (CSA) pertaining to the regulation of chemical mixtures which contain any of 34 listed chemicals. The NPRM was published to implement CSA requirements that only those chemical mixtures identified by regulation be exempt from applicable regulatory controls. The NPRM proposed criteria for the determination of whether a chemical mixture shall qualify for automatic exemption from CSA regulatory controls. Additionally, the NPRM defined an application process by which manufacturers may apply for an exemption for chemical mixtures that do not qualify for automatic exemption. Due to concerns regarding the potential illicit use of chemical mixtures which contain ephedrine, N methylephedrine, N- methylpseudoephedrine, norpseudoephedrine, phenylpropanolamine, and/or pseudoephedrine (as precursor material for the production of methamphetamine and related amphetamines), DEA is hereby finalizing the portion of the NPRM pertaining to these six chemicals. Final regulations for all remaining listed chemicals will be published under separate rulemaking, upon completion of a thorough review of applicable comments. PMID- 12725240 TI - Current awareness in prenatal diagnosis. PMID- 12725241 TI - Synthesis and conformational characterization of peptides related to the neck domain of a fungal kinesin. AB - The Y362K mutation in the neck domain of conventional kinesin from Neurospora crassa provokes a significant reduction of the rate of movement along microtubules. Since the alpha-helical coiled-coil structure of the neck region is implicated in the mechanism of the processive movement of kinesins, a series of peptides related to the heptad region 338-379 of the wild-type and the variant fungal kinesinswere synthesized as monomers and as N-terminal disulfide dimers, crosslinked to favour self-association into coiled-coil structures entropically. A comparison of the dichroic properties of the peptides and the effects of trifluoroethanol and peptide concentration clearly confirmed the strong implication of the single point mutation in destabilizing the intrinsic propensity of the peptides to fold into the supercoiled conformation. That there is a correlation between the stability of the coiled-coil and rate of movement of the kinesin is confirmed. PMID- 12725242 TI - Effects of detergents on the secondary structures of prion protein peptides as studied by CD spectroscopy. AB - Pathogenic prion proteins (PrP(Sc)) are thought to be produced by alpha-helical to beta-sheet conformational changes in the normal cellular prion proteins (PrP(C)) located solely in the caveolar compartments. In order to inquire into the possible conformational changes due to the influences of hydrophobic environments within caveolae, the secondary structures of prion protein peptides were studied in various kinds of detergents by CD spectra. The peptides studied were PrP(129-154) and PrP(192-213); the former is supposed to assume beta-sheets and the latter alpha-helices, in PrP(Sc). The secondary structure analyses for the CD spectra revealed that in buffer solutions, both PrP(129-154) and PrP(192 213) mainly adopted random-coils (approximately 60%), followed by beta-sheets (30%-40%). PrP(129-154) showed no changes in the secondary structures even in various kinds of detergents such as octyl-beta-D-glucopyranoside (OG), octy-beta D-maltopyranoside (OM). sodium dodecyl sulfate (SDS), Zwittergent 3-14 (ZW) and dodecylphosphocholine (DPC). In contrast, PrP(192-213) changed its secondary structure depending on the concentration of the detergents. SDS, ZW, OG and OM increased the alpha-helical content, and decreased the beta-sheet and random-coil contents. DPC also increased the alpha-helical content, but to a lesser extent than did SDS, ZW, OG or OM. These results indicate that PrP(129-154) has a propensity to adopt predominantly beta-sheets. On the other hand, PrP(192-213) has a rather fickle propensity and varies its secondary structure depending on the environmental conditions. It is considered that the hydrophobic environments provided by these detergents may mimic those provided by gangliosides in caveolae, the head groups of which consist of oligosaccharide chains containing sialic acids. It is concluded that PrP(C) could be converted into a nascent PrP(Sc) having a transient PrP(Sc) like structureunder the hydrophobic environments produced by gangliosides. PMID- 12725243 TI - Synthesis and use of a pseudo-cysteine for native chemical ligation. AB - The process of native chemical ligation (NCL) is well described in the literature. An N-terminal cysteine-containing peptide reacts with a C-terminal thioester-containing peptide to yield a native amide bond after transesterification and acyl transfer. An N-terminal cysteine is required as both the N-terminal amino function and the sidechain thiol participate in the ligation reaction. In certain circumstances it is desirable, or even imperative, that the N-terminal region of a peptidic reaction partner remain unmodified, for Instance for the retention of biological activity after ligation. This work discusses the synthesis of a pseudo-N-terminal cysteine building block for incorporation into peptides using standard methods of solid phase synthesis. Upon deprotection, this building block affords a de facto N-terminal cysteine positioned on an amino acid sidechain. which is capable of undergoing native chemical ligation with a thioester. The syntheses of several peptides and structures containing this motif are detailed, their reactions discussed. and further applications of this technology proposed. PMID- 12725244 TI - Conformational features of a synthetic model of the first extracellular loop of the angiotensin II AT1A receptor. AB - The angiotensin II AT1A receptor belongs to the G-protein coupled receptors (GPCRs). Like other membrane proteins, GPCRs are not easily amenable to direct structure determination by the currently available methods. The peptide encompassing the putative first extracellular loop of AT1A (residues Thr88 Leu100, el1) has been synthesized along with a cyclic model where the linear peptide has been covalently linked to a template designed to keep the distance between the peptide termini as expected in the receptor. The conformational features of the two molecules have been studied using circular dichroism and NMR techniques. The region W94PFG97 forms a type-II beta-turn and undergoes a Trp-Pro peptide bond cis-trans isomerization in both peptides confirming that these characteristics are intrinsic to el1. In addition, the presence of the spacer seems to modulate the flexibility of the peptide. PMID- 12725245 TI - The chemical synthesis and binding affinity to the EGF receptor of the EGF-like domain of heparin-binding EGF-like growth factor (HB-EGF). AB - Heparin-binding EGF-like growth factor (HB-EGF), which belongs to the EGF-family of growth factors, was isolated from the conditioned medium of macrophage-like cells. To investigate the effect of N- and C-terminal residues of the EGF-like domain of HB-EGF in the binding affinity to the EGF receptor on A431 cell. We synthesized HB-EGF(44-86) corresponding to the EGF-like domain of HB-EGF and its N- or C-terminal truncated peptides. Thermolytic digestion demonstrated three disulfide bond pairings of the EGF-like domain in HB-EGF is consistent with that of human-EGF and human-TGF-alpha. HB-EGF(44-86) showed high binding affinity to EGF-receptor, like human-EGF. The truncation of the C-terminal Leu86 residue from HB-EGF(44-86), HB-EGF(45-86) or HB-EGF(46-86) caused a drastic reduction in the binding affinity to the EGF receptor. These results suggest that the EGF-like domain of HB-EGF plays an important role in the binding to the EGF receptor, and its C-terminal Leu86 residue is necessary for binding with the EGF-receptor. In addition, the deletion of the two N-terminal residues (Asp44-Pro45) from HB EGF(44-86) caused a 10-fold decrease in relative binding affinity to the EGF receptor. This indicates that the two N-terminal residues of the EGF-like domain of HB-EGF are necessary for its optimal binding affinity to the EGF receptor. PMID- 12725246 TI - A rose is a metaphor is a remedy. PMID- 12725247 TI - Weighing the homeopathic evidence. PMID- 12725248 TI - Does homeopathy reduce the cost of conventional drug prescribing? A study of comparative prescribing costs in general practice. AB - There is little research on cost-effectiveness of homeopathy in General Practice. This study aimed to compare the costs of homeopathic prescribing with conventional drugs prescribing. Data were collected for 4 years on all patients who were treated homeopathically. Costs of homeopathic remedies and costs of conventional drugs which otherwise would be prescribed for these patients was calculated for the total duration of treatment. Savings were calculated. One hundred patients were included in the study. Average cost savings per patient was pounds 60.40. The majority of patients had improved and most did not report any side-effects. The limitations of this study are that it is based on one GP's work, with a small number of patients so definite and generalisable conclusions cannot be drawn. Moreover, calculated costs in this study are based on drugs only, it does not take into account doctor's time, special investigations and time off sick. Future work needs to be carried out to include all of these points for a comprehensive economic analysis. PMID- 12725249 TI - Homeopathic remedies as metaphors in family therapy. A narrative-based approach to homeopathy. AB - OBJECTIVE: To examine a metaphorical, narrative model to explain the outcome of an homeopathic, encounter involving the patient, practitioner and the homeopathic medicine. MATERIAL AND METHODS: The transcript of a videotaped doctor-patient interaction from a European doctor-patient communication study (EUROCOM) was analysed using qualitative methods. RESULTS: Analysis demonstrates a narrative relationship between the protagonists in the therapeutic interaction, with the homeopathic medicine performing a metaphorical role. DISCUSSION: The results justify further investigation of the narrative and metaphorical properties of the therapeutic encounter in homeopathy. PMID- 12725250 TI - The research evidence base for homeopathy: a fresh assessment of the literature. AB - BACKGROUND: The claims made for the clinical effects of homeopathy are controversial. The results of several meta-analyses of clinical trials are positive, but they fail in general to highlight specific medical conditions that respond well to homeopathy. AIMS: This review examines the cumulative research from randomised and/or double-blind clinical trials (RCTs) in homeopathy for individual medical conditions reported since 1975, and asks the question: What is the weight of the original evidence from published RCTs that homeopathy has an effect that is statistically significantly different from that in a comparative group? METHOD: Analysis of the 93 substantive RCTs that compare homeopathy either with placebo or another treatment. RESULTS: 50 papers report a significant benefit of homeopathy in at least one clinical outcome measure, 41 that fail to discern any inter-group differences, and two that describe an inferior response with homeopathy. Considering the relative number of research articles on the 35 different medical conditions in which such research has been carried out, the weight of evidence currently favours a positive treatment effect in eight: childhood diarrhoea, fibrositis, hayfever, influenza, pain (miscellaneous), side effects of radio- or chemotherapy, sprains and upper respiratory tract infection. Based on published research to date, it seems unlikely that homeopathy is efficacious for headache, stroke or warts. Insufficient research prevents conclusions from being drawn about any other medical conditions. CONCLUSIONS: The available research evidence emphasises the need for much more and better-directed research in homeopathy. A fresh agenda of enquiry should consider beyond (but include) the placebo-controlled trial. Each study should adopt research methods and outcome measurements linked to a question addressing the clinical significance of homeopathy's effects. PMID- 12725251 TI - Homeopathic aggravations: a systematic review of randomised, placebo-controlled clinical trials. AB - Homeopathic aggravations have often been described anecdotally. However, few attempts have been made to scientifically verify their existence. This systematic review aimed at comparing the frequency of homeopathic aggravations in the placebo and verum groups of double-blind, randomised clinical trials. Eight independent literature searches were carried out to identify all such trials mentioning either adverse effects or aggravations. All studies thus found were validated and data were extracted by both authors. Twenty-four trials could be included. The average number of aggravations was low. In total, 50 aggravations were attributed to patients treated with placebo and 63 to patients treated with homoeopathically diluted remedies. We conclude that this systematic review does not provide clear evidence that the phenomenon of homeopathic aggravations exists. PMID- 12725252 TI - Antibiotics and the development of resistant microorganisms. Can homeopathy be an alternative? AB - Antibiotic resistance is a global public health problem. Once confined primarily to hospitals it is now increasingly common in primary care. The prevalence of resistant bacteria is rising, and organisms resistant to almost all antibiotics have been identified. The main causes are indiscriminate prescribing and the use of antibiotics in animal feeds and other agricultural applications. Policies to restrict use of antibiotics have had limited success. Homeopathy may have a role to play in combating the development of antibiotic resistance. Clinical research suggests that homeopathy is effective in the treatment of upper respiratory tract infections in children, a frequent cause of inappropriate antibiotic prescribing. The EU recommends the use of homeopathy in organic animal husbandry, and it is used by significant numbers of farmers. At present there is little data on the effectiveness of homeopathy in this setting. Further research should be done. PMID- 12725253 TI - Homeopathic treatment of Japanese patients with intractable atopic dermatitis. AB - The objective of the study was to evaluate the efficacy of homeopathic treatment of intractable atopic dermatitis (IAD). Seventeen IAD patients were given individualized homeopathic treatment in addition to conventional dermatological therapy from 6 months to 2 years and 7 months. Although all of the patients had previously been treated with conventional medicine and various psychological approaches, they had had severe conditions and shown no significant sign of improvement. The efficacy of homeopathic treatment was measured by objective assessments of the skin condition and the patients' own assessments, using a 9 point scale similar to the Glasgow Homeopathic Outcome Scale, was used. Over 50% improvement was reported in overall impression and in their skin conditions by all patients, in itchiness by 15 of the patients, in sleep disturbance by 10 out of 13 patients, in satisfaction in daily life by nine out of 12, in fulfillment at work by seven out of 11 and in satisfaction with human relations by 10 out of 14. Two detailed case histories are reported. PMID- 12725254 TI - 20 years ago: British Homoeopathic Journal, April 1983. PMID- 12725255 TI - Homeopathy and the placebo effect. PMID- 12725257 TI - Is the 'rule of halves' in hypertension still valid?--Evidence from the Chennai Urban Population Study. AB - AIM OF THE STUDY: The aim of the present study was to assess the applicability of the rule of halves in an urban population in South India. METHODS: The Chennai Urban Population Study (CUPS) is an ongoing population based study involving two residential colonies representing a middle and lower socio-economic status in Chennai city. The response rate for the study was 90.1%. Based on a glucose tolerance test (GTT), the study subjects were categorized as having normal glucose tolerance test (NGT), impaired glucose tolerance test (IGT) or diabetes. Blood pressure was measured for all the study subjects and the subjects were classified as hypertensive using the criteria, systolic blood pressure (SBP) > or = 140 mm Hg and/or diastolic blood pressure > or = 90 mmHg and/or known hypertensives and/or treatment with antihypertensive drugs. Controlled hypertension was defined as SBP < 140 mmHg and DBP < 90 mmHg. RESULTS: The overall prevalence of hypertension in this population is 22.1% (279/1262). Of these 279 individuals with hypertension, only 37.3% (104/279) were known hypertensives. Of the 104 known hypertensives, only 52 subjects (50%) were under any kind of antihypertensive therapy. Of these 52 individuals, only 21 (40%) had blood pressure under control. Prevalence of hypertension was higher in subjects with glucose intolerance. Awareness and treatment of hypertension was slightly higher among the diabetic hypertensive subjects. Analysis on the different social classes revealed that the prevalence, awareness and treatment of hypertension to be significantly higher among the middle income group compared to lower income group. CONCLUSION: The rule of halves is still valid in this urban South Indian population and thus the awareness, treatment and control measures for hypertension are still inadequate in this population. PMID- 12725256 TI - Control of blood pressure in India: rule of halves still very much valid. PMID- 12725258 TI - Role of splenectomy in chronic idiopathic thrombocytopenic purpura. AB - AIM OF THE STUDY: To evaluate the usefulness of splenectomy and factors which predict long term remission in chronic idiopathic thrombocytopenic purpura (ITP). METHODS: We reviewed the data of 364 patients diagnosed as chronic ITP between January 1983 to December 1996 of whom 71 patients underwent splenectomy. The patients were followed up for an average period of 58 months and the short and long term response to splenectomy were analyzed at the end of one month and 60 months, respectively. RESULTS: At the end of one month after splenectomy, 82% had complete response, 7% partial response and 11% had no response. At the end of 60 months, 42% maintained complete response, 7% partial response, 34% had no response and 17% were lost to follow up. The results were statistically evaluated by using non-parametric test (Chi-square test) to age, sex, platelet count prior to treatment, initial response to steroids, time interval between diagnosis and splenectomy and post-operative platelet count. Of these factors only preoperative response to steroids (p value = 0.018303) and postoperative platelet count (p value = 0.013536) were found to be significant, statistically to predict long term remission. Age, sex, initial platelet count and time interval between diagnosis and splenectomy didn't seem to be statistically significant. CONCLUSION: This study suggests, that patients with an initial complete response to steroids and a post-operative platelet count > 300 x 10(9)/L at the time of discharge were associated with a long term remission. Splenectomy in ITP is a safe procedure with minimal morbidity and mortality and gives a good long term remission in steroid- failure patients with chronic ITP. PMID- 12725259 TI - Trial of heparin in viper bites. AB - AIMS OF THE STUDY: Viper bites produce hematotoxicity and coagulopathy which may be either true disseminated intravascular coagulation (DIC) or DIC-like syndrome. Role of heparin is studied in the present study of viper bite cases as use of heparin provides a rational therapy for defibrination caused by viper envenomation. METHODOLOGY: One hundred and twenty two patients with viper bite and incoagulable blood were randomised into test group and control group. Test group received heparin in addition to antisnake venom (ASV). Efficacy was assessed by monitoring bleeding time (BT), clotting time (CT), prothrombin time (PT), platelets, fibrinogen, blood urea (BU), serum creatinine, development of complications and overall outcome. RESULTS: Heparin group showed favourable outcome in all parameters studied. But many of them were not statistically significant. CONCLUSION: Heparin seems to be having a beneficial role which needs to be confirmed by larger trials and longer duration of heparin administration. PMID- 12725260 TI - Peripheral nerve complications in hemophilia. AB - AIM: Muscle bleed in hemophilics can result in compressive neuropathy and permanent disability if not treated properly in time. Twenty cases of peripheral nerve involvement in hemophilics are presented with an aim to highlight its features and the role of pulsed ultrasound in the management. METHODOLOGY: Between January, 1999 and 2000 June, 134 patients of bleeding disorders were examined. Twenty of them presented with features of nerve palsy. In addition to usual conservative treatment, pulse ultrasound electrotherapy of 10-15 sittings was used in all the cases. RESULTS: Femoral nerve was involved in majority (15), followed by sciatic (4) and peroneal nerve in one case. Femoral nerve involvement was associated with iliacus bleed in 13 patients and inguinal bleed in two cases. All the four cases of sciatic nerve palsy were associated with gluteal hematoma. Only case of peroneal nerve palsy was associated with hematoma at upper leg and calf. Twelve cases improved completely within six months whereas another four recovered around 10 months. Remaining four showed only partial recovery even when followed up to 12 months. CONCLUSIONS: Recognition of nerve complications in hemophilics is important to avoid long-term disability. Conservative therapy starting early was associated with favourable results. Pulsed ultrasound therapy at the site of haematoma helped in early regression of hematoma and neural recovery. PMID- 12725261 TI - Comparison of oral versus intravenous iron therapy in predialysis patients of chronic renal failure receiving recombinant human erythropoietin. AB - OBJECTIVES: To compare oral versus intravenous iron in pre-dialysis patients of chronic renal failure (CRF) receiving recombinant human erythropoietin (rHuEPO). METHODS: The study was undertaken in 40 adult patients of chronic renal failure. The patients were randomly divided into two groups A and B of 20 patients each. Group A patients were given oral iron and group B patients were given intravenous iron. All patients in both groups were given recombinant human erythropoietin 2000 IU twice weekly subcutaneously. The study was carried for up to three months. Patients were monitored every month for renal parameters and haematological parameters which included haemoglobin, reticulocyte count and packed cell volume. Ferrokinetic studies were done at baseline and at three months. RESULTS: It was observed that haematological parameters showed significant statistical improvement in the intravenous iron group as compared to group A (oral iron group). The ferrokinetic studies revealed that serum iron, serum ferritin and transferrin saturation, decreased significantly in oral iron group, whereas significant increase was seen in group B (intravenous iron group). None of the patients developed any adverse effects because of erythropoietin or iron therapy. CONCLUSIONS: Concomitant use of intravenous iron is better than oral iron in CRF patients treated with rHuEPO. The intravenous route of iron administration may be a preferred route along with rHuEPO therapy, more so in the Indian context where prevalence of iron deficiency anaemia is fairly high. PMID- 12725262 TI - Prevalence of renovascular abnormality in patients undergoing cardiac catheterization. AB - OBJECTIVE: To determine the prevalence of renal artery disease and to correlate the underlying risk factors like age, sex, diabetes, hypertension, urea and creatinine in patients who have undergone angiogram for cardiovascular diseases. METHODS: Retrospective analysis of the reports of angiogram of patients who have undergone cardiac catheterization in Vijaya Heart Foundation for cardiovascular diseases. RESULTS: The prevalence of renovascular stenosis is 12.4%. Prevalence of hypertension and diabetes in the group of patients with renovascular stenosis compared with group having coronary artery disease is not statistically significant (p > 0.8). Univariate and multivariate logistic identified age, diabetes, hypertension and urea as independent predicators of renal artery stenosis; while variables like sex and serum creatinine were not associated. CONCLUSION: High prevalence of unsuspected renovascular abnormalities is found in patients who undergo angiography for cardiovascular disease. Factors like age, diabetes, hypertension and urea could be clinical predicators of renal artery stenosis. Hence renal arteries should be visualized routinely in patients undergoing coronary angiogram for cardiovascular disease. PMID- 12725263 TI - Sudden onset visual impairment and deafness in a patient with "long standing rheumatoid arthritis". PMID- 12725264 TI - Wilson's disease. PMID- 12725265 TI - Polymyositis with arthritis. PMID- 12725266 TI - Isolated splenic hydatid cyst. PMID- 12725267 TI - Diagnosis and treatment of Indian visceral leishmaniasis. PMID- 12725268 TI - The muscular dystrophies--from genes to proteins. PMID- 12725269 TI - Outcome of pregnancies in renal allograft recipients. AB - Despite documented success and long term safety of thousands of pregnancies in female renal allograft recipients in Western countries, pregnancy is still a rare event, and considered risky in India in these patients. Four initial cases with their adverse outcome in the Indian context are presented. PMID- 12725270 TI - Postpartum posterior leukoencephalopathy syndrome. AB - In this report we are presenting three patients of posterior leukoencephalopathy syndrome developing in postpartum period. Two of these patients had persistent imaging abnormalities in posterior parietal and occipital regions leading to focal atrophy of brain along with permanent cortical blindness and recurrent seizures. In both the patients the syndrome was either unrecognized, or remained untreated on initial presentation. In third patient also the syndrome was not recognized for 10 days, initial clinical manifestations and computed tomographic (CT) abnormalities remained unchanged even after two months. Failure to early recognition and treatment can produce permanent brain damage and syndrome of posterior leukoencephalopathy may become irreversible. PMID- 12725271 TI - Chronic myeloid leukemia presenting with avascular necrosis of femur head. AB - We report the case of a 15 years girl in whom avascular necrosis (AVN) of right femur head was the presenting feature of chronic stable phase of chronic myeloid leukemia (CML). To date, only three case of CML with AVN have been reported. So, in view of rarity of this condition, a similar case of CML presenting as AVN of femur head is being reported. PMID- 12725272 TI - Amiodarone-induced systemic lupus erythematosus. AB - Drug-induced lupus is not a very common entity. Drugs like hydralazine, procainamide have been reported to induce lupus, but so far there is no case report of amiodarone-induced lupus in Indian literature. A case of atrial fibrillation in an elderly man who came with systemic features of lupus after amiodarone therapy for one and half years is reported. PMID- 12725273 TI - Reversible segmental portal hypertension--an unusual presentation of abdominal tuberculosis in a renal transplant recipient. AB - Infections are the commonest cause of morbidity and mortality in renal transplant recipients. In India, tuberculosis is a one such common infection in these patients and presents with protean manifestations. We report here a case of pyrexia of unknown origin (PUO) and segmental portal hypertension in a renal transplant recipient. Search for the cause of portal hypertension revealed abdominal tubercular lymphadenitis. Treatment with anti-tubercular therapy caused regression of segmental portal hypertension. PMID- 12725274 TI - Intra-arterial thrombolysis in acute brainstem infarct. AB - A young male presented with vertebrobasilar ischaemic stroke. He was given intra arterial thrombolysis, following which he showed gradual partial recovery. PMID- 12725275 TI - Acute inflammatory demyelinating polyneuropathy with P. falciparum malaria. AB - Various types of neuropsychiatric manifestations are described in P. falciparum malaria of which peripheral neuropathy has been described mainly from India. We are reporting such a case who presented with seven days history of fever and weakness of two days duration. On investigations it turned out to be acute inflammatory demyelinating polyneuropathy (AIDP) with peripheral blood showing heavy parasitaemia of P. falciparum. All other causes of acute polyneuropathy were ruled out by history and relevant examination. Patient improved with quinine and other supportive therapy. PMID- 12725276 TI - Acute intermittent porphyria: an unusual cause of malignant hypertension. AB - Hypertension is a rare complication of acute intermittent porphyria (AIP) and is related to the sympathetic over-activity seen in this condition. We report a patient with AIP with malignant hypertension that recurred with a subsequent episode. Mechanisms of hypertension and renal damage are discussed. PMID- 12725277 TI - Gastric lipoma presenting with dyspepsia. AB - Gastric lipoma is one of the rare benign gastric tumors. Its preoperative diagnosis obviates the need of an extended gastrectomy. We report a case of gastric lipoma who presented with symptoms of dyspepsia and was treated by surgical gastrectomy and tumour enucleation. PMID- 12725278 TI - Disseminated tuberculosis presenting as hemobilia, successfully treated by arterial embolization. AB - Tuberculosis, specially disseminated tuberculosis, involves the liver frequently. Focal hepatic tuberculosis with local hemorrhage has been reported. We report on a twenty-one year female with disseminated tuberculosis presenting with initially non-localisable massive upper gastrointestinal bleeding, subsequently found to have pancreatitis, right sided pleural effusion and hemobilia which was treated successfully. PMID- 12725279 TI - Presumptive artemether resistance in a patient with mixed malarial infection. PMID- 12725280 TI - Pulmonary nocardial infection with Pseudomonas infection of the tongue in patient with dermatomyositis. PMID- 12725281 TI - Outbreak of polyarthritis with pyrexia in Western Rajasthan. PMID- 12725282 TI - Nonfulminant subacute pulmonary fat embolism following fracture of radius and ulna. PMID- 12725283 TI - Extramedullary plasmacytoma associated with multiple myeloma. PMID- 12725284 TI - IgA myeloma presenting as diabetes mellitus with refractory anaemia. PMID- 12725285 TI - Typhoid hepatitis. PMID- 12725286 TI - Structure and biological activities of beta-glucans from yeast and mycelial forms of Candida albicans. AB - We have achieved the extraction of cell wall beta-glucan from the mycelial form of Candida albicans (C. albicans) IFO 0579 (M-CSBG) by using acetic acid, sodium hypochlorite (NaClO), and dimethylsulfoxide (DMSO) treatments. The yield of M CSBG was significantly lower (7.5% from dried mycelial cells) than that of the yeast form from C. albicans IFO 1385 (Y-CSBG, 25.9% from dried yeast cells). The properties of M-CSBG were similar to those of Y-CSBG in terms of nuclear magnetic resonance (NMR) spectra and limulus reactivity. Molecular weight (Mw) of M-CSBG was slightly higher than that of Y-CSBG. Both Y-CSBG and M-CSBG induced the production of comparable amounts of macrophage inflammatory protein-2 (MIP-2), a chemotactic factor, from mouse peritoneal exudate cells (PEC) in vitro. These findings suggest that the structure and properties of CSBG from yeast and mycelial cells are similar to each other. PMID- 12725287 TI - The first isolation of ustilaginomycetous anamorphic yeasts, Pseudozyma species, from patients' blood and a description of two new species: P. parantarctica and P. thailandica. AB - The genus Pseudozyma is ustilaginomycetous anamorphic yeasts, and are mainly isolated from plants. We isolated three Pseudozyma strains from the blood of patients in Thailand. While one isolate was identified as P. antarctica by rDNA sequence analysis, the other two were considered to be new species and were named P. parantarctica and P. thailandica. The three isolates proved to be resistant to 5-flucytosine, and P. thailandica was also resistant to fluconazole and itraconazole. As far as we know, this is the first isolation of Pseudozyma strains from humans. The two new species are described. PMID- 12725288 TI - Coenzyme Q1-catalyzed luminol chemiluminescent assay for rapid antimicrobial susceptibility testing of Mycobacterium bovis. AB - Coenzyme Q1 is herein proposed as the best catalyst among coenzymes Q and vitamins K for quinone-catalyzed luminol chemiluminescent assays applied to rapid determination of viability or rapid antimicrobial susceptibility tests of Mycobacterium bovis. Luminol chemiluminescence intensity (LCI) was determined 10 min after the incubation of M. bovis with coenzyme Q1, and was proportional to CFU (colony-forming unit)/ml in the range of 9,000 to 2,250,000. LCI depended on the the production of the superoxide anion (O2-) rather than H2O2 during a 10-min incubation of M. bovis with coenzyme Q1, as superoxide dismutase reduced LCI more effectively than catalase. The minimal inhibitory concentrations (MICs) of 10 kinds of antituberculous agents estimated on the basis of decrease in LCI after one or two days' cultivation were in good agreement with MICs determined by turbidity analysis, which requires upwards of 1 week to complete. PMID- 12725289 TI - Induction of apoptosis in macrophage cell line, J774, by the cell-free supernatant from Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa is able to secrete many virulence factors that are cytotoxic towards eukaryotic cells. To investigate the effect of the bacterium on macrophages, we obtained cell-free supernatants from P. aeruginosa (Pa) IID1117 (elastase-positive and protease-positive) and Pa IID1130 (elastase-positive and protease-negative). After 6 hr of incubation with the cell-free supernatant from the Pa IID1117 strain, the viability of J774 macrophages was shown to be significantly reduced (47.5+/-11%), but not Pa IID1130 (96.4+/-1.6%) at a concentration of 10% (v/v) compared to control J774 macrophages without any supernatant (97.2+/-1.7%) by the detection of trypan blue dye exclusion. The death of cells was further demonstrated to be due to apoptosis characterized by chromatin condensation and apoptotic bodies by Hoechst 33258 staining, DNA fragmentation by agarose gel electrophoresis and terminal deoxynucleotidyl transferase-mediated d-UTP nick end labeling (TUNEL). An activated subunit was found to be released from procaspase-3 in cell lysate. But in the presence of protease inhibitor, the apoptosis was completely blocked. The findings indicate that the Pa IID1117 strain is capable of inducing apoptosis in J774 macrophages. The apoptosis induced by the cell-free supernatant from Pa IID1117 strain is suggested to be dependent on protease, but not elastase. PMID- 12725290 TI - Effects of tungstosilicate on strains of methicillin-resistant Staphylococcus aureus with unique resistant mechanisms. AB - In a previous study, it was found that polyoxotungstates such as undecatungstosilicate (SiW11) greatly sensitized strains of methicillin-resistant Staphylococcus aureus (MRSA) to beta-lactams. In this report, the effects of SiW11 on several MRSA strains with unique resistant mechanisms were studied. SiW11 was still effective to MRSA mutants with higher beta-lactam resistance due to reduced cell-lytic activity. Since the antimicrobial effect of TOC-39 (a cephem antibiotic with strong affinity to penicillin-binding protein (PBP) 2') was not strongly enhanced in any case, it was confirmed that the sensitizing effect of SiW11 is due to reduced expression of PBP2'. However, the sensitizing effect of SiW11 was relatively weak in MRSA strains with lowered susceptibility to glycopeptide antibiotics. A certain resistant mechanism other than the mecA PBP2' system worked in such a strain. Interestingly, an MRSA mutant with the Eagle-type resistance was dramatically sensitized. This result suggests that SiW11 has another site of action besides reducing the expression of PBP2'. PMID- 12725291 TI - The role of glucan-binding proteins in the cariogenicity of Streptococcus mutans. AB - Streptococcus mutans produces glucan-binding proteins (Gbps), which appear to contribute to the virulence of S. mutans. GbpA and GbpC genes were inactivated by the insertion of antibiotic-resistant genes into each gbp gene of S. mutans MT8148 to generate Gbp-defective mutants. Sucrose dependent adherences of the GbpA- and GbpC-defective mutants were found to be significantly lower than those of their parent strains MT8148. Caries inducing activity of the mutants in rats was significantly lower than that of strain MT8148R (streptomycin-resistant strain of MT8148). These results suggest that GbpA and GbpC participate in cellular adherence to tooth surfaces and contribute to the cariogenicity of S. mutans. PMID- 12725292 TI - Correlation between the avidity maturation of anti-HSP70 IgG autoantibody and recombination activating gene expressions in peripheral lymphoid tissues of Toxoplasma gondii-infected mice. AB - The avidity maturation of anti-TgHSP70 IgG antibody produced by B-2 cells of BALB/c mice (a resistant strain) and that of anti-mHSP70 IgG autoantibody produced by B-1 cells of C57BL/6 mice (B6; a susceptible strain) was observed after Toxoplasma gondii infection. Recombination-activating genes (RAGs) were predominantly expressed in B-1 cells from peritoneal exudate cells (PECs) of T. gondii-infected B6 mice, while RAGs were expressed in B-2 cells from PECs of BALB/c mice. These results suggest that the involvement of RAG gene activations in the peripheral lymphoid tissues in the avidity maturation of anti-TgHSP70 IgG antibody and anti-mHSP70 IgG autoantibody in T. gondii-infected mice. PMID- 12725293 TI - The influence of ovine MHC class II DRB1 alleles on immune response in bovine leukemia virus infection. AB - We have reported previously that the alleles of the ovine leukocyte antigen (OLA) DRB1 gene that encode the Arg-Lys (RK) motif and the Ser-Arg (SR) motif at positions beta70/71 of the OLA-DRbeta1 domain are associated with resistance and susceptibility, respectively, to development of bovine leukemia virus (BLV) induced ovine lymphoma. Here, to investigate the different immune response in sheep that carried alleles associated with resistance and susceptible for 30 weeks after infection with BLV, we selected sheep that had the RK/RK or SR/SR genotype among the 52 sheep analyzed by polymerase chain reaction-restriction fragment length polymorphism and DNA sequencing of PCR product for the OLA-DRB1 exon 2 and infected them with BLV. Although the number of BLV-infected cells and virus titer had been maintaining low levels throughout the experimental period, the sheep with the RK/RK genotype could induce expansion of CD5- B-cells and rapid production of neutralizing antibody in the early phase of infection. The level of incorporation of [3H]thymidine by peripheral blood mononuclear cells from the sheep with RK/RK genotype gave a strong response to BLV virion antigen and synthetic antigenic peptides that corresponded to T-helper epitope of the BLV envelope glycoprotein gp51. In contrast, the sheep with SR/SR genotype showed a strong response to BLV virion antigen and synthetic antigenic peptides that corresponded to T-cytotoxic and B-cell epitopes. In such cases, the animals with the RK/RK strongly expressed IFN-gamma, the animals with SR/SR genotype strongly expressed IL-2. To determine the proliferating cells, we tried a blocking assay with monoclonal antibodies such as anti-CD4, -CD8 and -DR molecule. We found that these proliferating cells were MHC-restricted CD4+ T-cells. PMID- 12725294 TI - The urease gene cluster of Vibrio parahaemolyticus does not influence the expression of the thermostable direct hemolysin (TDH) gene or the TDH-related hemolysin gene. AB - In order to investigate why the thermostable direct hemolysin (TDH) and the TDH related hemolysin (TRH) of Vibrio parahaemolyticus are produced at low levels from urease-positive strains, the effect of the functional urease gene cluster of V. parahaemolyticus on the expression of the tdh and trh genes was examined. Transcriptional lacZ fusions with the tdh1, tdh2, trh1 and trh2 genes representing variants of the tdh and trh genes were integrated into the chromosome of an Escherichia coli strain and a urease-negative V. parahaemolyticus strain. The plasmid-borne urease gene cluster introduced and expressed in these constructs did not affect expression of any of the fusion genes. The amount of TDH produced from a Kanagawa phenomenon-positive V. parahaemolyticus did not change by introduction of the urease gene cluster either. It was concluded therefore that the urease gene cluster is not involved in the regulation of tdh and trh expression. PMID- 12725295 TI - Modulation of procarboxypeptidase R (ProCPR) activation by complementary peptides to thrombomodulin. AB - We designed complementary peptides (C-peptides) using a novel computer program (MIMETIC), which generates a series of peptides designed to interact with a target peptide sequence. Carboxypeptidase R (CPR) is an unstable basic carboxypeptidase found in fresh serum in addition to carboxypeptidase N (CPN) which is stable. CPR is generated from its precursor form (proCPR) by trypsin like enzymes, and its activation is mediated by thrombin generated in the coagulation cascade. The efficiency of activation is enhanced approximately 1,200 fold when thrombin (T) is bound to thrombomodulin (TM). We attempted to generate C-peptides which recognize the T-binding site within TM assuming that some of these might interfere with the generation of T and TM complexes (T-TM). Among three peptides designed, two inhibited the enhancement in activation of proCPR by T in the presence of TM. One of the peptides at 16 microM reduced the activation of proCPR to the level obtained by T alone. PMID- 12725296 TI - Chimeric human/murine monoclonal IgM antibodies to HIV-1 Nef antigen expressed on chronically infected cells. AB - Human IgM antibody (Ab) to gangliosides induced cytolysis of HIV-1-infected cells by homologous human complement. We expected that any human IgM Ab reactive with HIV-1 infected cells could cause complement-mediated cytolysis. The trans chromosome mouse (TC mouse) contains human chromosomes harboring genes responsible for immunoglobulin production. Spleen cells from TC mice immunized with recombinant Nef were fused with mouse myeloma cells to generate hybridomas, and we selected those that produced human mu-chain-positive Abs reactive with Nef fixed on an ELISA plate. However, the L-chain of the monoclonal Abs (mAbs) were murine lambda in type and were chimeric, and we could not succeed in obtaining mAb with human mu- and human kappa-chains. The chimeric mAbs reacted with the HIV 1 infected cells as seen with flow cytometric analysis, and the surface expression of Nef was also detectable on chronically infected OM10.1 cells which had no detectable gp120. However, although the reaction of the chimeric IgM mAb with HIV-1-infected MOLT4 cells induced C3 deposition on cell surfaces on incubation with fresh human serum, the cells remained unlysed, as determined by 51Cr release assay. The amount of Nef antigen on the cells might not have been high enough to overcome the function of HRF20 (CD59) that restricts formation of membrane attack complexes of homologous complement. However, combination of anti Nef IgM mAb with other IgM mAbs reactive with the surface of HIV-1-infected cells may induce a synergistic effect in complement mediated cytolysis. PMID- 12725297 TI - Military veterinarians engaged in Operation Iraqi Freedom. PMID- 12725298 TI - Bill would outlaw animal declawing in California. PMID- 12725299 TI - Lawmakers tackle animal hoarding. PMID- 12725300 TI - Beaver eager to be a conduit for "new and novel things". Interview by Susan C. Kahler. PMID- 12725301 TI - Animal tracking. PMID- 12725302 TI - Critical thinking. PMID- 12725303 TI - Suggests review of admission policies. PMID- 12725304 TI - More on testing for equivalence. PMID- 12725305 TI - Another solution to identify FIV-vaccinated cats. PMID- 12725306 TI - What is your diagnosis? Communication between the urethra and the corpus spongiosum, urethral stricture, mild cystitis, and presence of a urachal diverticulum. PMID- 12725307 TI - Effects of lufenuron treatment in cats on the establishment and course of Microsporum canis infection following exposure to infected cats. AB - OBJECTIVE: To determine effects of lufenuron treatment in cats on the establishment and course of Microsporum canis infection following exposure to infected cats. DESIGN: Experimental trial. ANIMALS: 24 healthy juvenile domestic shorthair cats. PROCEDURE: 8 cats were given lufenuron PO (133 mg/cat/mo, equivalent to a dose of 100 to 130 mg/kg [45 to 59 mg/lb] at the beginning of the study and 25 to 35 mg/kg [11 to 16 mg/lb] at the end of the study), and 8 were given lufenuron SC (40 mg every 6 months). The remaining 8 were used as untreated control cats. After 4 months, cats were challenged by the introduction of cats with mild, experimentally induced M canis infection into the rooms where cats were housed. Extent of resulting infections in the naive cats was monitored for 22 weeks by physical examination and fungal culture. RESULTS: All lufenuron treated and control cats became infected with M canis. Cats treated with lufenuron had significantly lower infection scores, compared with control cats, during the early weeks following exposure, and there was a more prolonged initial progression phase of the infection. Once infections reached peak intensity, they resolved over similar periods in lufenuron-treated and control cats. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that oral or SC administration of lufenuron to cats, at the dosages used and under the conditions of this study, did not prevent establishment of dermatophytosis following exposure to infected cats. Infection was established more slowly among cats treated with lufenuron, but once established, infection resolved in approximately the same amount of time in lufenuron-treated as in control cats. PMID- 12725308 TI - Comparison of results of three commercial heartworm antigen test kits in dogs with low heartworm burdens. AB - OBJECTIVE: To compare results of 3 commercial heartworm antigen test kits performed on serum samples from dogs infected with low numbers of adult female heartworms. DESIGN: Blinded laboratory evaluation. Sample Population-Serum samples from dogs (n = 208) proven at necropsy to be infected with 1 to 4 adult female heartworms and from dogs (32) without heartworms. PROCEDURE: Samples were sequentially tested with each test kit, following the manufacturers' instructions, by licensed veterinary technicians in private practice who were not aware of infection status of the dogs. The order of test kit evaluations was randomly chosen. For each test kit, sensitivity, specificity, accuracy, positive predictive value, and negative predictive value were evaluated. RESULTS: All tests yielded some false-negative results, and there were significant differences among tests in regard to ability to detect low heartworm burdens. Sensitivity of the test kits ranged from 78 to 84%. For all test kits, sensitivity increased as number of female heartworms increased. All 3 test kits had high specificity (97%). CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that sensitivity of the 3 commercially available heartworm antigen test kits ranged from 78 to 84% when used to test serum samples from dogs with low heartworm burdens, and that sensitivity varied among test kits. For all 3 test kits, specificity was 97%. All 3 test kits yielded false-positive and false-negative results for some dogs with low heartworm burdens. PMID- 12725309 TI - Clinical application of a polymerase chain reaction assay for diagnosis of leptospirosis in dogs. AB - OBJECTIVE: To evaluate the use of a polymerase chain reaction (PCR) assay on urine samples for diagnosis of leptospirosis in dogs. DESIGN: Prospective case study. ANIMALS: 132 dogs with clinical signs suggestive of leptospirosis and 13 healthy dogs. PROCEDURE: PCR testing was performed on urine samples to detect leptospiral DNA; results were compared with results of conventional criteria for the diagnosis of leptospirosis. RESULTS: Leptospirosis was diagnosed in 8 dogs via established criteria; all these dogs had positive results of PCR assay, including 1 dog with positive results before seroconversion developed. A positive PCR assay result was also obtained in 16 dogs that did not have a confirmed diagnosis of leptospirosis. In the 8 dogs that had a confirmed diagnosis of leptospirosis, serovars pomona (n = 3 dogs), grippotyphosa (2), canicola (2), and bratislava (1) were identified serologically. The remaining 121 dogs all had a diagnosis other than leptospirosis or were healthy. For PCR testing on urine, sensitivity was 100%, specificity was 88.3%, positive predictive value was 33%, and negative predictive value was 100%. CONCLUSIONS AND CLINICAL RELEVANCE: Positive PCR test results prior to seroconversion may have value in establishing an early diagnosis. Positive results in dogs that had signs consistent with leptospirosis despite failing to meet established criteria for leptospirosis raise questions regarding the sensitivity of serologic testing in diagnosis of leptospirosis. Serovars pomona, grippotyphosa, and canicola were most common. PMID- 12725310 TI - Comparison of polymerase chain reaction assay, bacteriologic culture, and serologic testing in assessment of prevalence of urinary shedding of leptospires in dogs. AB - OBJECTIVE: To compare results of polymerase chain reaction (PCR) testing of urine samples, serologic testing, and bacteriologic culture of urine to determine prevalence of urinary shedding of leptospires in dogs. DESIGN: Serial case study. ANIMALS: 500 dogs evaluated serially without regard to health status. PROCEDURE: Urine samples were examined via PCR assay and bacteriologic culture for leptospires. Blood samples were analyzed for antibodies against serovars canicola, bratislava, pomona, icterohemorrhagiae, grippotyphosa, and hardjo. RESULTS: Titers > or = 1:100 against at least 1 serovar were detected in 104 (20.8%) dogs, and titers > or = 1:400 were detected in 41 (8.2%) dogs. High titers were detected most commonly to serovar grippotyphosa, followed by icterohemorrhagiae, canicola, pomona, bratislava, and hardjo. High titers to > 1 serovar were detected in 14 dogs. A positive PCR assay result was obtained in 41 (8.2%) dogs, only 9 of which had a titer > or = 1:100. Leptospires were not cultured from the urine of any dog. Only 4 dogs had clinical leptospirosis. Overall disease prevalence was 0.8% for the 6-month evaluation period. Compared with PCR assay, serologic testing for predicting shedding had a sensitivity of 22%, specificity of 79%, positive predictive value of 9%, and negative predictive value of 92%. CONCLUSIONS AND CLINICAL RELEVANCE: Irrespective of health status, 8.2% of dogs were shedding pathogenic leptospires. Serologic testing was a poor predictor of urinary shedding. Clinically normal dogs that shed leptospires may pose a zoonotic risk to their owners. PMID- 12725311 TI - Association between lymph node size and metastasis in dogs with oral malignant melanoma: 100 cases (1987-2001). AB - OBJECTIVE: To determine the association between lymph node size and metastasis and to assess measurement of lymph node size as an accurate and reliable means of tumor staging in dogs with oral malignant melanoma. DESIGN: Retrospective study. ANIMALS: 100 dogs with histologically confirmed oral malignant melanoma. PROCEDURE: Clinical records for dogs with oral malignant melanoma were reviewed. Data regarding size and results of cytologic or histologic examination of lymph nodes were evaluated. The association between lymph node size and metastasis was determined. RESULTS: Forty-seven (47%) dogs, of which 23 (49%) had enlarged mandibular lymph nodes, had no cytologic or histologic evidence of metastasis. Of 53 (53%) dogs with cytologic or histologic evidence of mandibular lymph node metastasis, 37 (70%) had enlarged mandibular lymph nodes, and 16 (30%) had mandibular lymph nodes of normal size. Overall, 16 of the 40 (40%) dogs with normal-sized lymph nodes had microscopic evidence of metastatic disease. Sensitivity and specificity of lymph node size as a predictor of metastasis were 70 and 51%, respectively, and the positive and negative predictive values were 62 and 60%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Although a significant relationship was identified between lymph node size and metastasis to the lymph node, this association did not appear strong enough to be clinically relevant. Results suggest that lymph node size alone is insufficient for accurate clinical staging of oral malignant melanoma in dogs; cytologic or histologic examination of regional lymph nodes should routinely be performed, regardless of size of those nodes. PMID- 12725312 TI - Traumatic diaphragmatic hernia in cats: 34 cases (1991-2001). AB - OBJECTIVE: To determine whether signalment, duration of hernia, clinical signs, contents of hernia, CBC and serum biochemical abnormalities, concurrent injuries, perioperative treatment and administration of analgesics, results of intraoperative anesthetic monitoring data, or level of training of the veterinarian performing the herniorrhaphy was associated with mortality rate after surgical repair of traumatic diaphragmatic hernia in cats. DESIGN: Retrospective study. ANIMALS: 34 cats. PROCEDURE: Review of medical records and a telephone follow-up with owners and referring veterinarians were performed. RESULTS: Mean age of affected cats was 3.6 years; cats that survived to the time of discharge were significantly younger than cats that died or were euthanatized. Tachypnee was the most common clinical sign at hospital admission; cats that survived to the time of discharge had significantly higher respiratory rates than cats that died or were euthanatized after surgery. Postoperative complications developed in 50% of cats; tachypnea and dyspnea were most common. Mortality rate was not associated with duration of hernia or results of preoperative CBC and serum biochemical analyses, but was significantly associated with concurrent injuries. Mortality rate was not associated with hernia contents, intraoperative use of positive inotropes or corticosteroids, episodes of hypotension or severe hypoxia during anesthesia, or level of training of the veterinarian performing the surgery. CONCLUSIONS AND CLINICAL RELEVANCE: Cats that are older or have low to mildly increased respiratory rates and concurrent injuries are more likely to die after surgical repair of traumatic diaphragmatic hernia. PMID- 12725313 TI - West Nile virus encephalomyelitis in horses: 46 cases (2001). AB - OBJECTIVE: To determine signalment, clinical findings, results of diagnostic testing, outcome, and postmortem findings in horses with West Nile virus (WNV) encephalomyelitis. DESIGN: Retrospective study. ANIMALS: 46 horses with WNV encephalomyelitis. PROCEDURE: Clinical data were extracted from medical records of affected horses. RESULTS: On the basis of clinical signs and results of serologic testing, WNV encephalomyelitis was diagnosed in 46 of 56 horses with CNS signs. Significantly more males than females were affected. Increased rectal temperature, weakness or ataxia, and muscle fasciculations were the most common clinical signs. Paresis was more common than ataxia, although both could be asymmetrical and multifocal. Supportive treatment included anti-inflammatory medications, fluids, antimicrobials, and slinging of recumbent horses. Results of the IgM capture ELISA and the plaque reduction neutralization test provided a diagnosis in 43 horses, and only results of the plaque reduction neutralization test were positive in 3 horses. Mortality rate was 30%, and 71% of recumbent horses were euthanatized. One horse that had received 2 vaccinations for WNV developed the disease and was euthanatized. Follow-up communications with 19 owners revealed that most horses had residual deficits at 1 month after release from the hospital; abnormalities were resolved in all but 2 horses by 12 months after release. CONCLUSIONS AND CLINICAL RELEVANCE: Our findings were similar to those of previous WNV outbreaks in horses but provided additional clinical details from monitored hospitalized horses. Diagnostic testing is essential to diagnosis, treatment is supportive, and recovery rate of discharged ambulatory horses is < 100%. PMID- 12725314 TI - Ultrasonographic findings in horses with right dorsal colitis: five cases (2000 2001). AB - OBJECTIVE: To determine whether ultrasonography would be useful in the diagnosis of right dorsal colitis in horses. DESIGN: Retrospective study. ANIMALS: 5 horses with right dorsal colitis and 15 healthy adult horses. PROCEDURE: Mural thickness and appearance of the right dorsal colon were determined from ultrasonographic images obtained at right intercostal spaces 10, 11, 12, 13, and 14. RESULTS: The right dorsal colon could be imaged most consistently at the right 11th, 12th, and 13th intercostal spaces, below the margin of the lung and axial to the liver. Mural thickness measured from ultrasonographic images was significantly greater in horses with right dorsal colitis than in healthy horses. The right dorsal colon in affected horses had a prominent hypoechoic layer associated with submucosal edema and inflammatory infiltrates. Successful treatment of 1 horse with right dorsal colitis was associated with a decrease in mural thickness coincident with an increase in serum albumin and total protein concentrations and weight gain. A decrease in mural thickness was also observed in a second horse treated for right dorsal colitis that was not associated with healing of the right dorsal colon or an increase in serum albumin concentration but rather thinning of a segment of the right dorsal colon that eventually ruptured. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that ultrasonographic measurement of mural thickness and evaluation of the appearance of the right dorsal colon may be useful in the diagnosis of right dorsal colitis in horses. PMID- 12725315 TI - Evaluation of cattle for experimental infection with and transmission of Brucella suis biovar 4. AB - OBJECTIVE: To determine whether cattle can become persistently infected with Brucella suis biovar 4, whether the organism can be transmitted vertically or horizontally, and whether tests for bovine brucellosis are diagnostic. DESIGN: Observational study. ANIMALS: 24 pregnant cows and their calves and 6 bulls. PROCEDURE: Cows and bulls were housed separately in groups of 6 with each group consisting of 3 cattle experimentally infected with B suis biovar 4 and 3 naive animals. Cattle were observed for clinical signs daily; blood samples were collected weekly. Clotted blood from each sample was submitted for bacterial culture. Serum was tested with an indirect ELISA and the standard tube agglutination test (STAT), buffered plate agglutination test, brucellosis card test (BCT), and complemen't fixation test (CFT). Tissues collected at necropsy were submitted for bacterial culture and histologic examination. RESULTS: All 15 inoculated cattle seroconverted on 2 or more serologic tests, and bacteria were isolated from 4 inoculated cows at necropsy. There was no bacteriologic evidence of vertical or horizontal transmission, and none of the cattle developed clinical abnormalities or gross or histologic lesions. Results of the indirect ELISA were positive for all inoculated cattle. The other tests gave variable results; the CFT, STAT, and BCT yielded negative results for at least 1 of the 4 cattle from which the organism was isolated. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that cattle-to-cattle transmission of B suis biovar 4 is unlikely. Serologic tests for bovine brucellosis should be used cautiously when attempting to identify cattle with rangiferine brucellosis, as they do not discriminate between the 2 diseases and vary in their ability to detect exposed cattle. PMID- 12725316 TI - Primary endocrinomas (carcinoids and variant neoplasms) of the gallbladder. A statistical evaluation of 138 reported cases. AB - This study was carried out to obtain extensive information on carcinoids (the carcinoid group) and related variant endocrinomas (the variant group) of the gallbladder, and to statistically analyze their characteristics from various clinicopathologic aspects. A total of 138 cases were collected from the international sources, 101 belonging to the carcinoid group and 37 to the variant group. The first group consisted of 81 cases of typical and 20 atypical carcinoids. Comparative evaluation was attempted mainly between the carcinoid and variant groups, and occasionally between the typical and atypical carcinoid series when statistical significance was suspected. The carcinoid group showed a statistically significant difference from the variant group by exhibiting a younger average age (61.7 years vs 69.7 years: P<0.01), a higher incidence of associated cholelithiasis (87.3% vs 56.0%: P<0.01), a higher incidence of small tumors 50 mm or less (85.2% vs 52.9%: P<0.01), a smaller average tumor-size (29.6 mm vs 58.7 mm: P<0.01), a lower rate of metastases (40.7% vs 70.6%: P<0.05), a higher immunoreactivity rate of chromogranin (100.0% vs 66.7%: P<0.01), a lower immunoreactivity rate of gastrin (23.8% vs 70.6%: P<0.01), and a higher five-year survival rate (60.4% vs 21.3%: P<0.0005). Significant differences in various clinicopathological aspects confirmed between the carcinoid group and the variant group suggested that endocrine carcinomas of these two groups perform a different clinical pattern, represented most clearly by postoperative outcomes. These groupings are decided on the basis of histologic patterns, namely, well to poorly differentiated endocrine carcinomas (typical to atypical carcinoids) and undifferentiated or anaplastic variants of other endocrine carcinomas. The basic criteria for such classification of these endocrine carcinomas based on international agreements are required. PMID- 12725317 TI - Prevention of radiotherapy-induced emesis. AB - In this minireview the authors examine and discuss the radioprotective compounds and the new combination therapies for the prophylaxis of radiation-induced emesis. Radiation-induced emesis is an important secondary effect of this anticancer treatment and it represents one of the causes of therapy interruption and decay of life quality before the introduction of optimal control of radiation induced emesis with new antiemetic drugs which ensure the continuance of radiotherapy and avoid time breaks, that could negatively influence the efficacy of anticancer treatment. The incidence, the severity or the latency of radiotherapy-induced nausea and vomiting are correlated both with the treatment features (fractions, total dose, irradiation site) and with the main clinical characteristics of the patients. In contrast to the very extensive literature on the prevention of chemotherapy-induced emesis, relatively few studies about the prevention of nausea and vomiting in patients submitted to radiotherapy have been published. Among antiemetic drugs for the prevention of emesis, benzamides and in particular metoclopramide, are widely used in clinical practice. The introduction of selective 5-HT3 antagonists in clinical practice produced an important improvement in control of chemotherapy induced emesis, but few published studies were aimed at evaluating the efficacy of these drugs in the prophylaxis of nausea and vomiting due to radiation exposure. We herewith present a brief summary of Clinical practice guidelines for the use of antiemetics in anticancer therapy recently published by ASCO (American Society of Clinical Oncology). PMID- 12725318 TI - Pregnancy-associated breast cancer: a case control and long-term follow-up study in China. AB - Pregnancy associated (PA) breast cancer is defined when diagnosed during pregnancy or within one year afterwards. To analyze the prognostic factors related to this disease and assess the impact of pregnancy on breast cancer patients, 88 PA breast cancer patients initially treated in the Shanghai Cancer Hospital from 1957 to 1990 were reviewed. A non-PA group including 176 patients individually well matched to the PA group was also analyzed. Univariate analysis suggested prognostic value for clinical tumor size, TNM stage, and breast feeding time among the classic prognostic factors, pregnancy associated factors and treatment modalities were evaluated. Multivariate analysis demonstrated clinical tumor size, TNM stage and axillary lymph node metastasis as independent prognostic factors. Compared with the non-PA group, the PA group was significantly correlated with delay at diagnosis, large size of the tumor, late TNM stage, extension to the skin or chest wall and administration with oophorectomy. The overall survival rates of 5-year, 10-year and 20-year were 40.39%, 36.29% and 30.70%, respectively, which were worse than those in the non PA group, but did not reach a significant difference (p=0.0536). We are conducting further basic research to analyze the biologic characteristics of PA breast cancer. PMID- 12725319 TI - Prognostic factors affecting survival in male breast cancer. AB - Male breast cancer is an uncommon disease and most of the data concerning its treatment and prognostic factors have been obtained from studies spanning a long period of time during which the main concepts of treatment in breast cancer have changed. In the current study, patients affected with male breast cancer treated in the last decade with a multidisciplinary approach were reported from a single Institution. Fifty-five patients with male breast cancer treated between 1990 and 1998 were included in this study. The median follow-up period was 36 months. Five year disease-free and overall survival were 44.5% and 72.9%, respectively. Patients' age, tumor size, axillary status, tumor type and treatment modalities such as surgery, chemotherapy, radiotherapy, and hormonotherapy were evaluated as possible prognostic factors affecting the disease-free and overall survival periods. In univariate analysis, tumor size, axillary status and extent of surgery were found to be affecting overall survival significantly whereas axillary status, tumor type, extent of surgery and chemotherapy affected disease free survival. In multivariate analysis, axillary status, extent of surgery and tumor type were found as independent prognostic factors for overall survival and radiotherapy was added to these factors for disease-free survival. In conclusion, the approach to male breast cancer patients should be similar to that for female patients. Modified radical mastectomy should be the gold standard followed by combined adjuvant therapy individualized for patients due to axillary status, tumor size and type. PMID- 12725320 TI - CT cystoscopy in the evaluation of bladder tumors. AB - The aim of the present study was to assess the role of virtual cystoscopy in the identification of bladder tumors. Fifteen patients (11 men and 4 women, median age: 61 years, range: 46-74 years) with a positive finding of bladder tumor at fiber-optic cystoscopy were studied by multislice-CT. Scans were downloaded to a workstation with the aid of a software for the processing of 3-D reconstructions, with a volume-rendering technique which allowed the "navigation" within the bladder in search of wall lesions. In this group of 15 patients, cystoscopy was able to detect 19 neoplastic lesions, 13 with a diameter >10 mm and 6 with a diameter <10 mm. Virtual cystoscopy, instead, identified 17 lesions (89%) only. In particular, all those lesions with a diameter >1 cm (13/13=100%) were correctly identified, whereas only 4 of the 6 lesions with a diameter <1 cm were depicted. The 2 false negative cases were 2 lesions with a flat morphology, measuring 5 and 6 mm. Most recent technological advances allowed the employement of virtual endoscopies, characterized by the absence of invasivity as compared with fiber-optic studies and based on data obtained by spiral- and multislice CTs. According to our experience, virtual CT-cystoscopy revealed to be a complementary tool in the evaluation of cross-sectional images and proved to be an easy procedure without complications, well-accepted by the patients, and with a reliable detection of those bladder lesions measuring more than 5 mm in case of polypoid formations and at least 10 mm in case of flat lesions. This technique, however, does not allow the collection of a bioptic sample and--with the present resolution power of available equipments--it could be unable to correctly detect small-sized flat lesions. We, nonetheless, believe that this procedure, in the future, thanks to rapid technological improvements in virtual imaging techniques, could become a useful diagnostic tool in the management of those patients with bladder tumors. Further studies on larger study groups are therefore desirable for a more reliable validation of the technique. PMID- 12725321 TI - Chemoprevention for colorectal tumorigenesis associated with chronic colitis in mice via apoptosis. AB - The mechanism of the suppressive effect of nonsteroidal anti-inflammatory drugs in azoxymethan and dextran sulfate sodium-induced colonic aberrant crypt foci/tumors associated with chronic colitis in mice was studied. With administration of sulindac, a cyclooxygenase-1 and -2 inhibitor, the mean number of colonic aberrant crypt foci/tumors was significantly smaller than that of controls. There was no significant difference in prostaglandin E2 content in the colonic mucosa between the groups. Furthermore, nimesulid, a cyclooxygenase-2 selective inhibitor, also suppressed colonic aberrant crypt foci/tumors as well as sulindac. Administration of nimesulid caused apoptosis indices to be significantly higher along with cyclooxygenase-2 expression being significantly lower than in controls. Apoptosis indices of 400 ppm group of nimesulid were significantly higher than that of 200 ppm group. Nonsteroidal anti-inflammatory drugs distinctly suppress the occurrence of aberrant crypt foci/tumors in this murine colitis-associated neoplasia model. Induction of apoptosis is a more important factor for chemoprevention than this reduction of prostaglandin E2. PMID- 12725322 TI - Genetic differences in interleukin-1 betapolymorphisms among four Asian populations: an analysis of the Asian paradox between H. pylori infection and gastric cancer incidence. AB - The prevalence of Helicobacter pylori infection is high among Asian populations, but the incidences of gastric cancer differ greatly among northern and southern Asian populations. Here, we studied histopathological findings in stomach tissue using an updated Sydney System and the frequencies of interleukin (IL) 1betapolymorphisms, thought to be associated with an increased risk of gastric cancer, in four Asian populations. Endoscopic-guided biopsies from three regions of the stomach and the -511 T-to-C polymorphism in the IL-1betagene were examined in 228 Japanese, 116 Chinese, 159 Thai and 83 Vietnamese patients with gastric diseases. H. pylori colonization, inflammation and activity were more severe in the Japanese and Thai populations than in the Chinese and Vietnamese populations and these scores were more antrum-predominant in the Thai and Vietnamese populations than in the Japanese and Chinese populations, with the most severe degree of atrophy and intestinal metaplasia occurring in the angulus region of the Japanese population. The IL-1betapolymorphisms did not differ among the four populations overall, but in cases with severe mucosal atrophy (pepsinogen I/II ratio <3.0), the CC polymorphism was dominant in the Japanese population and the TT+TC polymorphism was dominant in the Chinese population; no difference in C and T allele frequencies was found in the Thai and Vietnamese populations. In conclusion, the incidence of gastric cancer is extremely low, but the prevalence of H. pylori infection is high in the Thai population (Asian paradox). In the Thai population, the scores for corpus gastritis and intestinal metaplasia, which are associated with a high risk of gastric cancer, were low in comparison with the Japanese population. IL-1betapolymorphisms were correlated with mucosal atrophy in the Japanese and Chinese populations, but not in the Thai and Vietnamese populations. PMID- 12725323 TI - Transcriptional activation of cyclin-dependent kinase inhibitor, p21waf1 gene by treatment with a differentiation inducing agent, vesnarinone in a human salivary gland cancer cell line. AB - Recently, a new concept for cancer therapy termed "tumor dormancy therapy" has been proposed. The concept of this therapy is to prolong the survival time of cancer patients while maintaining their quality of life. We have been developing a differentiation-inducing therapy, which is included in the tumor dormancy therapy, for salivary gland cancer. In this study, we examined the effect of a differentiation-inducing drug, Vesnarinone on the growth of several cancer cells, and examined the molecular mechanism by which Vesnarinone induces the cyclin dependent kinase inhibitor, p21waf1 in the cancer cells. Vesnarinone significantly suppressed the growth of TYS (salivary gland cancer cells), PC3 (prostate cancer cells), and A431 (squamous cell cancer cells). Furthermore, Vesnarinone dose-dependently enhanced the expression of p21waf1 mRNA in TYS cells. Using the luciferase reporter assay it was found that the enhancement of p21waf1 mRNA expression by Vesnarinone was through direct transcriptional activation of the p21waf1 promoter. Thus, analyzing the molecular mechanisms of differentiation inducing drugs may lead to the development of a new therapeutic strategy for several human malignancies, including salivary gland cancer. PMID- 12725324 TI - Arsenic trioxide induces apoptosis of rat hepatocellular carcinoma cells in vivo. AB - The aim of this work was to study the effect of arsenic trioxide (As2O3) on rat hepatocellular carcinoma (HCC), and investgate on the mechanisms of its antitumor effect. HCC was induced by chemocarcinogen diethylnitrosamine (DEN) in Wistar rats, that were then treated with As2O3 intraperitoneally in three different concentrations once a day for two weeks, and twice a week for another two weeks. The histological and ultrastructural changes in liver tissue were observed under microscope and electronic microscope on the 7th, 14th and 28th day after drug administration. The apoptosis and cellular dynamic parameters of tumor cells were observed by flow cytometry. The expression of bcl-2, bax, and proliferation cell nuclear antigen (PCNA) of rat liver cancer cells on the 7th day after drug administration was determined by using immunohistochemical technique. Treatment with As2O3 caused HCC cells death via both apoptotic and non-apoptotic mechanisms when the dose was high (5 mg.kg(-1)), while necrosis was rare and apoptosis was common when the dose was appropriate (1 mg.kg(-1)). This effect was obviously accompanied with accumulation of cells in G2/M phases (G2/M restriction). Many apoptotic cells were also found in G2/M phases. The expression intensity of bcl-2 or bax varied depending on the dose administrated. Downregulation of bcl-2/bax was observed, accompanied with upregulation of apoptosis. However, the ratio of bcl-2/bax and the percentage of apoptosis were not the utmost when the dose administered was the highest. In conclusion, these data demonstrate that As2O3 induces apoptosis of rat HCC cells, and it is closely associated with G2/M restriction when apoptosis reaches the top. Apoptosis can be observed in all three phases of cell cycle, but it is more common in G2/M phase when the dose is appropriate. It is suggested that arsenic trioxide may be an atypical cell cycle specific agent. Apoptosis of tumor cells is closely associated with down regulation of the ratio of bcl-2/bax, but that may not be the only dominant factor. PMID- 12725326 TI - Studies on the role of some synthetic curcuminoid derivatives in the inhibition of tumour specific angiogenesis. AB - In this study, some of the synthetic curcuminoid derivatives are analyzed for their anti-angiogenic activity. Intraperitoneal administration of the compounds tetrahydrocurcumin (THC), salicyl curcumin (SC) and curcuminIII (C-III) reduced the number of tumour directed capillaries induced by injecting B16F-10 melanoma cells on the ventral side of C57BL/6 mice. THC (14.5 +/- 2.5 capillaries) and SC (16 +/- 2.5 capillaries) were more significant (P < 0.001) than C-III (19 +/- 1.8 capillaries) compared to the untreated control (30.8 +/- 2.1 capillaries). Treatment with these curcuminoids reduced the serum NO as well as TNF-alpha levels of the angiogenesis-induced animals. In vitro NO and TNF-alpha production by the activated MOs were reduced in a concentration dependent manner by the treatment of the curcuminoid compounds. PMID- 12725325 TI - Transfer of IkappaBalpha gene increase the sensitivity of paclitaxel mediated with caspase 3 activation in human lung cancer cell. AB - In oncogenic therapies, apoptosis seems to be the important mechanism of deciding chemotherapy effect. NF-kappaB transcription factors are implicated in the control of cell proliferation and apoptosis. NF-kappaB is activated by chemotherapy and by irradiation, and this pathway has been shown to protect cells potently from their stimuli-induced apoptosis. Furthermore, inhibition of NF kappaB leads to enhanced apoptosis in response to various stimuli. However, because the role of NF-kappaB as a modifier of the intrinsic chemosensitivity of cancer cells is less clear, we have studied the impact of IkappaBalpha (an inhibitor of NF-kappaB) on the chemosensitivity of human lung cancer cells. We used adenoviral vectors expressing human IkappaBalpha (AdIkappaBalpha) and investigated the effects of IkappaBalpha gene transfer in combination with 6 anticancer agents on a human pulmonary adenocarcinoma cell line, A549. Solutions containing anticancer agents at various concentrations were added followed by the addition of recombinant adenovirus solutions, and each IC50 was calculated based on the dose-response curves. The gene transfer of AdIkappaBalpha decreased IC50 from 12.0 to 2.2 nM on paclitaxel and increased IC50 from 0.27 to 16.0 microM on SM5887 compared with the transfer of control gene, AdLacZ. The IC50 did not change clearly on the other anticancer drugs. To investigate this molecular mechanism, we measured caspase 3 activity by the transfer of IkappaBalpha gene. On result, paclitaxel increased caspase 3 activity and SM5887 decreased the activity. These results indicate that the cell killing effect of anticancer drug is influenced by the inhibition of NF-kappaB activity and may, at least in part, depend on the regulation of caspase 3 activation. Adenovirus mediated IkappaBalpha gene transfer improve the anti-cancer effect of paclitaxel to lung cancer cells through the regulation of caspase 3 activation. PMID- 12725327 TI - The effects of dexamethasone on leukemic cells derived from patients with AML. AB - This study was performed to investigate the effects of dexamethasone on acute myelocytic leukemia (AML) cells. The study has been designed to investigate the in vitro effects of dexamethasone on the proliferative capacity, maturation and apoptosis of leukemic cells derived from patients with AML. Dexamethasone induced leukemic cell apoptosis and decreased cell count compared to the control. The percentage of cellular maturation increased in the dexamethasone groups compared to the control groups. Proliferative capacity of the cells was similar in all groups. These effects of dexamethasone on leukemic cells may be related to activation of the genes which stimulate apoptosis and maturation. More comprehensive studies are needed to evaluate the apoptotic effect of dexamethasone on AML cells. PMID- 12725328 TI - The clinical significance of sialyl Lewis antigen expression in the spread of gastric cancer. Flow cytometric DNA analysis. AB - We previously reported significant relationships between sialyl Lewis antigen expression on gastric cancer cells and both hepatic metastasis and clinical prognosis. The purpose of this study was to compare the expression of sialyl Lewis antigens on gastric cancer cells to elucidate the possible role of sialyl Lewis antigens in predicting the spread of a tumor with regard to histological findings. Subjects consisted of 38 patients with gastric cancer. For comparison we measured the values for sialyl Lewisa (sLea) and sialyl Lewisx (sLex) expression on the surface of about 10,000-30,000 cancer cells. Monoclonal antibodies CA19-9 and KM-93 were used to determine the frequency (%) and quantity (channel) of the expression by flow cytometry. We assessed the correlation of sLea and sLex expression with histological findings (depth of tumor invasion (pT), lymphatic invasion (ly), venous invasion (v), and lymph node metastasis (pN)), by comparing sLea and sLex expression in relation to the grade of histological findings. A significant relationship was found between lymphatic invasion and the frequency of sLea expression (r = 0.40, p<0.05). The mean values of sLea frequency in cases categorized as ly 2 (36.30) and ly 3 (31.81) were statistically higher than those in ly 1 cases (12.74) (p<0.05). A significant relationship was also observed between lymph node metastasis and the frequency of sLea expression (r = 0.46, p<0.01). The mean value of sLea frequency in pN 3 cases (44.14) was statistically higher than those in pN 0 (17.11) and pN 1 (19.03) cases (p<0.05). Neither the depth of tumor invasion nor venous invasion showed any correlation with the expression of sialyl Lewis antigens. In conclusion the frequency of sLea expression on the surface of gastric cancer cells was greater in those patients who developed lymphatic invasion and lymph node metastasis. However, the mechanism by which sialyl Lewisa expression is upregulated remains unclear. PMID- 12725329 TI - p53 mutations detection in urinary bladder cancer in the Greek population: application of the NIRCA assay. AB - The aim of the present study was to assess the use of an alternative molecular approach for p53 mutation detection and to investigate the usefulness of p53 as a prognostic indicator in bladder cancer. We applied the NIRCA assay, which consists of two-step PCR amplification, transcription of the amplified sequence, hybridisation of the transcripts and treatment with RNAses which recognizes mismatches due to the presence of mutations. Results of molecular analysis are correlated with immunohistochemical findings, standard clinopathological parameters and survival. p53 mutations were detected in 42.4% of the 66 examined TCCs cases. We could not demonstrate any statistical relationship between the presence of p53 mutation and p53 protein overexpression, and tumor stage or grade. A trend towards higher mutation rate in higher grade tumours was observed, although this failed to reach statistical significance. Despite the observation that the alterations of p53 gene are associated features of aggressive phenotype of transitional cell carcinomas they do not seem to offer additional prognostic information. PMID- 12725330 TI - Usefulness of analysis of p53 alteration and observation of surface microstructure for diagnosis of ulcerative colitis-associated colorectal neoplasia. AB - Patients with ulcerative colitis (UC) have a higher incidence of colorectal cancer. UC-associated colorectal cancer is thought to develop in patients with preexisting UC-associated dysplasia. It is crucial to diagnose UC-associated dysplasia and early stage of cancer in patients with long-standing UC for the purpose of treatment of UC-associated neoplasia. However, it is difficult to detect UC-associated dysplasia and the early stage of cancer endoscopically, and to discriminate these neoplasias from inflammatory regenerative epithelium pathologically. The aim of this study was to clarify whether observation of the surface microstructure could aid in the detection of UC-associated neopalsia, and whether analysis of genetic alterations could be used to discriminate between UC associated neoplasia and inflammatory regenerative epithelium. Tissue samples were obtained from colectomy specimens from eight cases of UC-associated neoplasia. We examined the surface microstructure of these tissues using stereomicroscopy. We also investigated mutation of K-ras codon 12 using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and alteration of the p53 gene, using immunohistochemistry and PCR-single stranded conformation polymorphism (PCR-SSCP). The surface microstructure of UC-associated neoplasia revealed a packed distribution of oval and/or, club-shaped and/or, branch-shaped pits and a villous appearance. Nuclear accumulation of p53 protein occurred in 59.5% of UC-associated neoplasia. Mutations of the p53 exon 5-8 were detected in 95.2% of UC-associated neoplasia, and these mutations were detected in 92.9% of UC-associated neoplasia that showed negative p53 immunohistochemical staining. Mutations of the p53 exon 5-8 in regenerative epithelium occurred infrequently. The K-ras mutation rate in UC-associated neoplasia was 7.4%. In conclusion, immunohistochemistry and PCR-SSCP analysis of p53 would be useful tools for pathological discrimination between UC-associated neoplasia and inflammatory regenerative epithelium, and observation of the surface microstructure may contribute to accurate endoscopic detection of UC-associated neoplasia. PMID- 12725331 TI - Dedifferentiation and decreased expression of adhesion molecules, E-cadherin and ZO-1, in colorectal cancer are closely related to liver metastasis. AB - Carcinoma cells with high metastatic potential often show a dedifferentiated phenotype at the primary site. In this study, a total of 48 cases (24 primary tumors of colorectal cancer (Pr-CRC) with liver metastasis, 24 without) were examined for E-cadherin and ZO-1 expression by immunohistochemical staining, and for their dedifferentiated phenotype. The expression levels of E-cadherin and ZO 1 were markedly decreased in the cancer cells of tumors with liver metastasis. Moreover, dedifferentiation of cancer cells, which was evaluated by the modified Gleason score, was also related to liver metastasis. However, none of the conventional clinicopathologic parameters of invasion, except lymph node metastasis, showed any relationship with liver metastasis. These results indicate that dedifferentiation and a decreased expression of E-cadherin and ZO-1 are closely related to liver metastasis. PMID- 12725332 TI - Correlation of gangliosides GM2 and GM3 with metastatic potential to lungs of mouse B16 melanoma. AB - Ten B16 mouse melanoma cell lines with increasing metastatic potential to lungs (B16LuF1 to B16LuF10) were generated by in-vitro & in-vivo selection technique starting with B16F1 melanoma cell line. The number of metastatic tumor nodules in lungs rose with increasing metastatic potential. Tumor cell gangliosides of B16LuF1 to B16LuF10 cell lines, analysed and compared with TLC, showed eight major ganglioside bands. Band1 to band6 corresponded with standard gangliosides GT1b, GD1b, GD1a, GM1, GM2 and GM3 respectively. Band7 and Band8 could not be identified. The concentration of total as well as individual ganglioside bands of B16LuF1 to B16LuF10 cells appeared to rise with increasing metastatic potential. Gangliosides from the plasma of these cell lines (B16LuF1 to B16LuF10) maintained in-vivo in C57BL/6 mice on TLC analysis gave eight major ganglioside bands, similar to those of cells. Plasma gangliosides appeared to rise with increasing metastatic potential. However, it was interesting to see that only band5 and band6 gangliosides in plasma increased almost linearly with increasing metastatic potential. The remaining six ganglioside bands in the plasma did not show such correlation. Band5 and Band6 gangliosides corresponded with standard gangliosides GM2 and GM3 respectively. Gangliosides of the spent culture media, secreted by these cell lines in-vitro in tissue culture also gave eight major ganglioside bands, similar to that of cells. Spent culture media gangliosides appeared to increase with increasing metastatic potential. However, concentration of only band5 and band6 gangliosides of spent culture media increased almost linearly with increasing metastatic potential, thus further confirming the role of band5(GM2) and Band6(GM3) gangliosides in regulating metastatic potential of B16 melanoma cells to lung. PMID- 12725333 TI - Analysis of lewis antigens on cell surface and alpha1,3 fucosyltransferase subtypes in H7721 human hepatocarcinoma cells. AB - The expressions of Lewis antigens on H7721 human hepatocarcinoma cells were detected with monoclonal antibodies and flow-cytometry. It was found that H7721 mainly expressed SLex, and a small amount of SDLex, but Lex and SLea was negligible. The monoclonal antibody of SLex (KM93) significantly blocked the adhesion of H7721 cells to human umbilical vein epithelial cells, as well as cell migration and invasion, but the blocking effect of SDLex antibody (FH6) was not statistically significant. The expressions of five subtypes of alpha1,3fucosyltransferases (alpha1,3FucTs), the enzyme responsible for the fucosylation step in Lewis antigen synthesis, were also studied using real-time RT-PCR. The expression of FucT mRNAs were FucT-IV > FucT-III > FucT-VI > or = FucT-VII > FucT-IX. FucT-VI is supposed to be the main enzyme responsible for the synthesis SLex and SDLex. FucT-VII and III may also participate in SLex synthesis. The absence of FucT-IX expression and FucT-III not being the rate limiting enzyme for SLea synthesis may be the reasons for the negligible expressions of Lex and SLea on the cell surface, respectively. PMID- 12725334 TI - Adrenal glands metastases from malignant melanoma. Laparoscopic bilateral adrenalectomy. AB - Adrenal metastases from Malignant Melanoma (MM) represent a debated therapeutical problem particularly in the case of disseminated disease. Surgical treatment, however, seems to be able to provide improvement on survival. Laparoscopic adrenalectomy is considered a gold standard procedure in benign adrenal disease but its value in malignancy, in terms of oncological effectiveness, is not known. A case of bilateral adrenal malignant melanoma metastases is reported. The patient, affected by superficial spreading melanoma of the right foot, eleven years after the primary developed a right adrenal metastasis. The relapse was treated by laparoscopic right adrenalectomy. One year later the patient had a new metastasis in the left adrenal gland and was submitted to laparoscopic left adrenalectomy. The two step laparoscopic bilateral adrenalectomy showed to be quite easy to perform, providing a complete removal of the whole glands, without adrenal tissue crushing and without neoplastic tissue dissemination in abdominal cavity. The postoperative course was excellent and the patient was discharged within about 72 hours after the two procedures. In literature only few reports indicate the feasibility of laparoscopic adrenalectomy for malignancy. In the reported case of malignant melanoma metastasis, minimally invasive adrenalectomy was very satisfactory and the good results obtained suggest its routine use. PMID- 12725335 TI - Gastroduodenal metastases in a clear cell sarcoma patient. AB - Clear cell sarcoma is a rare tumor with an unpredictable clinical course and usually poor prognosis. Almost all of the patients will have pulmonary metastases during the clinical course. Other more common metastatic sites are bones, lymph nodes, liver and spleen. Treatment of clear cell sarcoma is primarily surgical aiming to achieve uninvolved margins. Role of chemotherapy, radiotherapy and immunotherapy is controversial and remission of metastases has been reported rarely. In this study we report a case of clear cell sarcoma, with gastroduodenal and pulmonary metastases and remission of these metastases in response to chemotherapy. PMID- 12725336 TI - Subcutaneous metastasis of pancreatic cancer in the site of percutaneous biliary drainage. AB - A subcutaneous metastatic lesion from a carcinoma of the pancreas or common bile duct along the tract of a percutaneous transhepatic biliary drainage is a rare finding. Prompted by a case that came to our observation by chance, we reviewed the literature and analysed the 29 cases collected. Neoplastic cell seeding along a percutaneous drainage tract, albeit rare, must be kept in mind. The complication can be avoided if patients at risk, whenever possible, undergo endoscopic drainage. PMID- 12725337 TI - Bilateral irreversible hearing loss associated with the combination of carboplatin and paclitaxel chemotherapy: a unusual side effect. AB - The regimen with paclitaxel and platinum compound (carboplatin or cisplatin) is the standard chemotherapy for patients with advanced ovarian cancer. Ototoxity for carboplatin and paclitaxel alone or combined is rarely observed. We report the case of a 35-year old female with advanced ovarian cancer who developed sudden bilateral sensorineural hearing loss related to paclitaxel and carboplatin based chemotherapy. This uncommon adverse effect of carboplatin and paclitaxel alone or combined is discussed and the literature reviewed. Hearing monitoring should be mandatory to evaluate the real incidence of clinical and sub-clinical hearing modification induced by carboplatin and paclitaxel based chemotherapy. PMID- 12725338 TI - Studying health in Greenland: obligations and challenges. AB - Health research in Greenland has contributed with several findings of interest for the global scientific community and has documented health problems and risk factors of importance for planning the local health care system. The study of how health develops in small, scattered communities during rapid epidemiological transition carries prospects of global significance. The Inuit are a genetically distinct people living under extreme physical conditions. Their traditional living conditions and diet are currently undergoing a transformation, which may approach their disease pattern to that of the industrialized world, while still including local outbreaks of tuberculosis. Health research in Greenland is logistically difficult and costly, but offers opportunities not found elsewhere in the world. A long tradition of registration enhances the possibilities for research. A number of research institutions in Denmark and Greenland have conducted health research in Greenland for many years in cooperation with, among others, researchers in Canada and Alaska. National and international cooperation is supported by the Danish/Greenlandic Society for Circumpolar Health, the International Union for Circumpolar Health, and the Commission for Research in Greenland. Health news are regularly reported to international and local congresses and to the scientific journals. PMID- 12725339 TI - An overview of factors influencing the health of Canadian Inuit infants. AB - BACKGROUND: Inuit infants throughout the Arctic experience higher mortality and poorer health than their non-Inuit counterparts, and suffer disproportionately from bacterial and viral infections. STUDY DESIGN: This review examines the health status of these infants, with a focus on Canadian Inuit communities and reference to other circumpolar regions, as appropriate. It is based on a Medline search (1965 to present), special analyses of the 1996 Canadian Census and various national surveys, and selected government reports and documents. RESULTS: A wide range of inter-related factors affect the health of Inuit infants: their demographic, social, economic and physical environment, as well as personal health practices and the availability of high quality, culturally appropriate health services. Some of these factors may influence the susceptibility of Inuit infants to infection. Smoking is highly prevalent in Inuit communities, and its indisputable negative effects on health, including increased risk of respiratory tract infection in infants, represent an urgent public health challenge. CONCLUSION: Locally driven, focused and methodologically sound epidemiological research that addresses key gaps in knowledge could lead to more appropriate and effective preventive strategies to improve health in northern communities. PMID- 12725340 TI - Change in plasma levels of vitamin D after consumption of cod-liver and fresh cod liver oil as part of the traditional north Norwegian fish dish "Molje". AB - OBJECTIVE: To assess changes in plasma 25-hydroxy vitamin D (25(OH)D) concentrations after ingestion of <>, a traditional north Norwegian fish dish rich in vitamin D. METHODS: Thirty-three volunteers all living in the city of Tromso, located in northern Norway (latitude 690), were served a "Molje" meal consisting of cod, hard roe, cod liver, and fresh cod-liver oil. The amounts of liver, and cod-liver oil consumed were weighed and recorded. Blood samples were collected before the meal, and at 4 hours, 12 hours and 5 days after it. The cod liver and cod-liver oil were analysed for vitamin D content and the plasma samples for the metabolite 25(OH)D. Trends in plasma 25(OH)D levels during the five-day observation period were analysed. The study was conducted at the beginning of April of 2000. RESULTS: Among the 33 participating subjects, 69.7% had baseline plasma 25(OH)D concentrations below 50 nmol/l and for one-quarter of the subjects, they were < 37.5 nmol/l The participants who acknowledged taking cod-liver oil supplements had significantly higher baseline 25(OH)D plasma levels at the outset of the study compared to those reporting not doing so (p = 0.02). Changes in plasma 25(OH)D levels relative to baseline concentrations were significantly associated with the body mass index (p < 0.01). CONCLUSION: Vitamin D status in populations living in circumpolar areas needs more research to investigate to what degree people living in the Arctic areas are at increased risk for vitamin D insufficiency and to determine the role of the traditional diet in preventing such deficiency. PMID- 12725341 TI - Indigenous Greenlanders have a higher sero-prevalence of IgG antibodies to Helicobacter pylori than Danes. AB - OBJECTIVE: To assess the sero-prevalence of IgG antibodies to Helicobacter pylori (H. pylori) in Greenlanders and compare with the sero-prevalence in Caucasian Danes. METHODS: 71 randomly recruited indigenous Greenlanders (29 men) with a median age of 39 years (range 22-76), living in the capital, Nuuk, and the town of Ilulissat, and participating in a population survey carried out in 1993-1994. The results were compared with those obtained in a 1983-1984 population survey in Copenhagen County, comprising 2794 Caucasian Danes (1425 men) with a median age of 41 years (range 30-60). Serum IgG antibody levels to H. pylori were determined by an indirect enzyme-linked immunosorbent assay in 1995 and categorised as negative, borderline (equivocal), or positive. RESULTS: Greenlanders: 48.3% of men and 45.2% of women had positive serum IgG antibody levels to H. pylori In the entire series, 46.5% had positive IgG antibody levels, 25.4% displayed borderline antibody levels and 28.2% had negative antibody levels. There were no age, or gender differences concerning the prevalences of the three H. pylori IgG antibody subgroups. Danes: 25.6% of men, and of women, had positive IgG antibody levels against H. pylori. In the entire series, 25.6% had positive IgG antibody levels, 19.0% displayed borderline antibody levels and 55.4% had negative antibody levels. There was no gender difference concerning the sero-prevalence of IgG antibodies, but the sero-prevalence increased significantly with age. The prevalence of positive serum IgG antibodies against H. pylori was markedly higher in Greenlanders than in Danes (p < 0.0001). CONCLUSION: Indigenous Greenlanders have a significantly higher infection rate with H. pylori than Danes. The results suggest that Greenlanders become infected with H. pylori early in life. PMID- 12725342 TI - The Harstad Injury Prevention Study. A decade of community-based traffic injury prevention with emphasis on children. Postal dissemination of local injury data can be effective. AB - OBJECTIVES: To evaluate the outcome of a community-based program for reducing traffic injury rates with special focus on children and to assess the impact of a Traffic Injury Report (TIR) in terms of awareness and attitudes about safety issues. SETTING: The Norwegian cities Harstad (23 000) and Trondheim (140 000), during ten years. METHODS: The outcome was evaluated using hospital-based injury recording. Sustainability of the prevention program was promoted by disseminating information on the community's traffic injury profile. Reports containing information about traffic injuries were distributed quarterly to all Harstad households, containing victim stories and statistics on medical data and the location of the accidents. The impact of the reports was evaluated, using a questionnaire mailed to persons 18-80 years old. RESULTS: From the first two years (mean rate 116.1/10,000 person years), to last two years, a significant 59% [confidence interval (CI): 42% to 71%] reduction of traffic injury rates was observed for Harstad children. Overall rates for all ages decreased 37% [CI:47% to 24%] in Harstad increased by 3% [CI:-4% to 10%] in Trondheim (reference city). Significantly higher scores were found in Harstad compared to Trondheim concerning the awareness of, and positive attitudes towards, safety issues (e.g. alcohol and driving, speeding and children's safety in traffic). 56.0% of respondents in Harstad reported having acquired information, or good advice, about traffic safety from the reports. CONCLUSIONS: Traffic injuries in children can be prevented by community-based interventions. Distributing written information may enhance the program's sustainability. PMID- 12725343 TI - Special school teachers require more goal-oriented collaboration with psychiatric professionals in Northern Finland. AB - BACKGROUND: The problems of children and adolescents have increased in number and severity during the last years. OBJECTIVE: The purpose of this study was to elicit the views of special school teachers in the Oulu Province in Finland concerning their pupils' problems and the need, use and adequacy of relevant psychiatric services. METHODS: The information was collected from the Northern Ostrobothnia Hospital District in 1998 in Finland and responses were obtained from the 37 (97.4%) special schools. The data were analysed using the content analysis method. RESULTS: Special-needs pupils had various behavioural and emotional problems that made it difficult for them to learn and for the teachers to teach. According to the teachers, the major obstacles in the process of helping special-needs pupils were the delayed admission for treatment and the lack of information necessary for the pupils' school work and goal-oriented aftercare, although there were also favourable experiences of functional co operation and availability of useful information. Some schools lacked a reliable network for helping pupilsand supporting teachers. The Finnish legislation on basic education obliges the providers of education to provide rehabilitation in connection with special education and to arrange relevant development, counselling and support services. CONCLUSION: The rehabilitation of special-need pupils and the collaboration between school and mental health authorities is not optimally realised in spite of the legislation. PMID- 12725344 TI - The Nordic Council for Arctic Medical Research. History, aims and achievements. AB - The Organization of Nordic Council for Arctic Medical Research (NCAMR, or NoSAMF in Scandinavian languages) was initiated by the governmental Nordic Council in 1966. The new council was charged with the task of promoting arctic medical research in the Nordic countries. It began its duties in 1969. Originally the council covered Denmark, Finland, Norway and Sweden; Iceland joined as a member in 1977. During the first years the NCAMR held two to three conferences a year, the proceedings of which were communicated in the Nordic Council for Arctic Medical Research Reports series, distributed in about 1500 copies, free of charge. In 1971, 1981, 1987 and 1993, the NCAMR hosted the International Congress on Circumpolar Health and played a pivotal role in the establishment of the International Union for Circumpolar Health (IUCH) in 1986. Thereafter, the activities of the NCAMR developed a much more international character. Accident prevention, cold research, pollution, family health and, in later years, the health of indigenous peoples, became priorities, along with the establishment of international research networks. The NCAMR's report series soon developed into an established international journal under the title Arctic Medical Research. The inter-governmental financial support to the NCAMR was discontinued at the end of 1996. Thereafter, the secretariat operated under the auspices of the University of Oulu. The International Journal for Circumpolar Health, as it was named from 1997, continued to flourish, being published by the IUCH, the Nordic Society for Circumpolar Health and the University of Oulu. PMID- 12725345 TI - Apparent transmission of two species of gram-negative rods in catheterized residents on a 50-bed nursing home unit. PMID- 12725346 TI - Molecular epidemiology of methicillin-resistant Staphylococcus aureus in a Veterans Administration Medical Center. PMID- 12725347 TI - A gown is a gown is a gown: or is it? PMID- 12725348 TI - Nosocomial infections in a Turkish university hospital: a 2-year survey. PMID- 12725349 TI - Nasal carriage of methicillin-resistant Staphylococcus aureus among healthcare workers of an Iranian hospital. PMID- 12725350 TI - More is more. PMID- 12725351 TI - Colonization and infection with multiple nosocomial pathogens among patients colonized with vancomycin-resistant Enterococcus. AB - OBJECTIVE: To test the hypothesis that patients colonized with vancomycin resistant Enterococcus (VRE) have a higher frequency of colonization or infection with other nosocomial pathogens than do patients who are not colonized with VRE. DESIGN: A rectal swab culture survey was conducted to determine the point prevalence of stool colonization with ceftazidime-resistant gram-negative bacilli in hospitalized patients with or without VRE stool colonization. For a 6-month period, the frequency of Clostridium difficile diarrhea and isolation of antibiotic-resistant (ie, ceftazidime-, piperacillin/tazobactam-, levofloxacin-, or trimethoprim/sulfamethoxazole-resistant) gram-negative bacilli, methicillin resistant Staphylococcus aureus (MRSA), and non-albicans Candida species from clinical specimens other than stool was examined. SETTING: A Department of Veterans Affairs medical center. PATIENTS: All patients hospitalized in the acute care facility and one nursing home unit during a 1-week period in February 2001. RESULTS: VRE-colonized patients had a higher point-prevalence of rectal colonization with ceftazidime-resistant gram-negative bacilli than did patients not colonized with VRE (17% vs 4%; P = .026). During a 6-month period,the VRE colonized patients were more likely to have Clostridium difficile-associated diarrhea (26% vs 2%; P = .001), MRSA infection (17% vs 4%; P = .017), or colonization or infection with gram-negative bacilli resistant to 4 different antibiotics. CONCLUSION: VRE-colonized patients in our institution have a higher frequency of colonization or infection with other nosocomial pathogens than do patients who are not colonized with VRE. This suggests that isolation measures implemented to control VRE could help limit the dissemination of other, coexisting pathogens. PMID- 12725352 TI - Natural history of colonization with vancomycin-resistant enterococci, methicillin-resistant Staphylococcus aureus, and resistant gram-negative bacilli among long-term-care facility residents. AB - OBJECTIVE: To determine the natural history of colonization with vancomycin resistant enterococci (VRE), methicillin-resistant Staphylococcus aureus (MRSA), and resistant gram-negative bacilli among long-term-care facility (LTCF) residents. DESIGN: Observational cohort study. SETTING: A 355-bed LTCF with a ventilator unit and a subacute unit. PARTICIPANTS: Residents with colonization or infection with VRE, MRSA, or resistant gram-negative bacilli housed at the LTCF between December 1, 1999, and February 29, 2000. METHODS: Cultures of clinical and surveillance sites were performed at regular intervals. Charts were reviewed for clinical characteristics associated with clearance of colonization. Kaplan Meier curves were constructed to analyze the number of days to clearance of colonization. RESULTS: Forty-nine residents had 65 episodes of colonization (27 VRE, 30 MRSA, and 8 resistant gram-negative bacilli). Eighteen (28%) of the episodes cleared. The clearance rate was 2.7 episodes per 1,000 person-days. Clearance occurred significantly more often with resistant gram-negative bacilli colonization compared with VRE or MRSA colonization (6 [75%] vs 12 [21%]; P = .007; relative risk, 4.17; 95% confidence interval, 1.26 to 11.8). There was a trend toward longer use of antimicrobial agents among residents with persistent colonization. Infections occurred most frequently with MRSA. The urinary tract was the most common site of infection. CONCLUSION: Among LTCF residents, colonization with resistant gram-negative bacilli is four times more likely to clear than colonization with VRE or MRSA. Performance of surveillance cultures at regular intervals may reduce the need for contact precautions for LTCF residents with resistant gram-negative bacilli colonization. PMID- 12725353 TI - Effect of nosocomial vancomycin-resistant enterococcal bacteremia on mortality, length of stay, and costs. AB - OBJECTIVE: To determine the impact of vancomycin-resistant enterococcal bacteremia on patient outcomes and costs by assessing mortality, excess length of stay, and charges attributable to it. DESIGN: A population-based, matched, historical cohort study. SETTING: A 1,025-bed, university-based teaching facility and referral hospital. PATIENTS: Two hundred seventy-seven vancomycin-resistant enterococcal bacteremia case-patients and 277 matched control-patients identified between 1993 and 2000. RESULTS: The crude mortality rate was 50.2% and 19.9% for case-patients and control-patients, respectively, yielding a mortality rate of 30.3% attributable to vancomycin-resistant enterococcal bacteremia. The excess length of hospital stay attributable to vancomycin-resistant enterococcal bacteremia was 17 days, of which 12 days were spent in intensive care units. On average, dollars 77,558 in extra charges was attributable to each vancomycin resistant enterococcal bacteremia. To adjust for severity of illness, 159 pairs of case-patients and control-patients, who had the same severity of illness (All Patient Refined-Diagnosis Related Group complexity level), were further analyzed. When patients were stratified by severity of illness, the crude mortality rate was 50.3% among case-patients compared with 27.7% among control-patients, accounting for an attributable mortality rate of 22.6%. Attributable excess length of stay and charges were 17 days and dollars 81,208, respectively. CONCLUSION: Vancomycin-resistant enterococcal bacteremia contributes significantly to excess mortality and economic loss, once severity of illness is considered. Efforts to prevent these infections will likely be cost-effective. PMID- 12725354 TI - The epidemiology of vancomycin-resistant Enterococcus colonization in a medical intensive care unit. AB - OBJECTIVE: To determine the epidemiology of colonization with vancomycin resistant Enterococcus (VRE) among intensive care unit (ICU) patients. DESIGN: Ten-month prospective cohort study. SETTING: A 19-bed medical ICU of a 1,440-bed teaching hospital. METHODS: Patients admitted to the ICU had rectal swab cultures for VRE on admission and weekly thereafter. VRE-positive patients were cared for using contact precautions. Clinical data, including microbiology reports, were collected prospectively during the ICU stay. RESULTS: Of 519 patients who had admission stool cultures, 127 (25%) had cultures that were positive for VRE. Risk factors for VRE colonization identified by multiple logistic regression analysis were hospital stay greater than 3 days prior to ICU admission (adjusted odds ratio [AOR], 3.6; 95% confidence interval [CI95], 2.3 to 5.7), chronic dialysis (AOR, 2.4; CI95, 1.2 to 4.5), and having been admitted to the study hospital one to two times (AOR, 2.3; CI95, 1.4 to 3.8) or more than two times (AOR, 6.5; CI95, 3.7 to 11.6) within the past 12 months. Of the 352 VRE-negative patients who had one or more follow-up cultures, 74 (21%) became VRE positive during their ICU stay (27 cases per 1,000 patient-ICU days). CONCLUSION: The prevalence of VRE culture positivity on ICU admission was high and a sizable fraction of ICU patients became VRE positive during their ICU stay despite contact precautions for VRE-positive patients. This was likely due in large part to prior VRE exposures in the rest of the hospital where these control measures were not being used. PMID- 12725355 TI - The changing epidemiology of vancomycin-resistant enterococci. AB - OBJECTIVE: To determine the distribution of vancomycin-resistant enterococci (VRE) cases in our hospital and those from outside of our hospital from 1993 through 1998. METHODS: Weekly rectal surveillance was instituted whenever there were two or more cases present in the units. Cases were divided into acquired in our hospital, acquired outside of our hospital (VRE positive after and within 72 hours of admission, respectively), and indeterminate. Hospital cases were attributed to the originating ward or intensive care unit (ICU) if patients were noted to be positive within 72 hours of transfer. RESULTS: From 1993 to 1998, the rate of VRE per 1,000 admissions increased threefold, from 3.2 to 9.8, for the hospital. VRE cases acquired outside of the hospital increased by approximately 5% per year (r = 0.87; P = .03). The rate of VRE per 1,000 admissions increased 1.7-fold in the ICUs and 3.6-fold in the wards. The ICUs had an average of 75.3 cases per year, with the number of new cases per year increasing by approximately 9 (r = 0.80; P = .028). In the wards, there were an average of 22.0 new cases per year, with a slight upward trend of 3 additional new cases per year (r = 0.69; P = .64). There was a highly significant increasing linear trend (P = .0007) for VRE colonization and infection. CONCLUSION: Although VRE still predominate in the ICUs, cases originating from outside of our hospital and the wards are becoming more common. VRE colonization remained more frequent than infection. PMID- 12725356 TI - Risk factors for Stenotrophomonas maltophilia bacteremia in oncology patients: a case-control study. AB - OBJECTIVE: To characterize risk factors for Stenotrophomonas maltophilia bloodstream infection in oncology patients. DESIGN: A 3:1 case-control study. SETTING: Stem Cell Transplant and Leukemic Center at Barnes-Jewish Hospital (St. Louis), a 1,442-bed, tertiary-care teaching hospital with a 26-bed transplantation ward. METHOD: From, June 1999 to April 2001, 13 patients with S. maltophilia bacteremia were compared with 39 control-patients who were on the transplantation unit on the same day as the case-patients' positive blood cultures. Information collected included patient demographics, medical history, history of transplantation, transplantation type, graft versus host disease, neutropenia, antibiotic use, chemotherapy, mucositis, diarrhea, the presence of central venous catheter(s), cultures, and concomitant infections. RESULTS: Significant risk factors for S. maltophilia bacteremia included severe mucositis (7 [53.8%] of 13 vs 8 [20.5%] of 39; P = .034), diarrhea (7 [53.8%] of 13 vs 8 [20%] of 39; P = .034), and the use of metronidazole (9 [69.2%] of 13 vs 8 [20.5%] of 39; P = .002). In addition, the number of antibiotics used (median, 9 vs 5; P < .001), duration of mucositis (median, 29 vs 15 days; P = .032), and length of hospital stay (median, 34 vs 22 days; P = .017) were significantly different between case- and control-patients. Nine S. maltophilia isolates tested by pulsed-field gel electrophoresis were found to be distinctly different. CONCLUSION: Interventions to ameliorate the severity of mucositis, reduce antibiotic pressure, prevent diarrhea, and promote meticulous central venous catheter care may help prevent S. maltophilia bloodstream infection in oncology patients. The role of gastrointestinal tract colonization as a potential source of S. maltophilia bacteremia in oncology patients deserves further investigation. PMID- 12725358 TI - Implementation of the Canadian contingency plan for a case of suspected viral hemorrhagic fever. AB - OBJECTIVE: To describe the implementation of the Canadian contingency plan for viral hemorrhagic fever (VHF) in response to a suspected case. SETTING: A 300 bed, tertiary-care, university-affiliated hospital. PARTICIPANTS: A 32-year-old Congolese woman admitted to the hospital with suspected VHF in February 2001. Contact evaluation included hospital healthcare workers and laboratory staff. INTERVENTION: Enhanced isolation precautions were implemented in the patient care setting to prevent nosocomial transmission. Contact tracing and evaluation of close and high-risk contacts with symptoms was conducted. Laboratory precautions included barrier precautions and diversion of specimens. Communication occurred media. to both hospital employees and the media. RESULTS: Three high-risk contacts, 13 close contacts, and 60 casual contacts were identified. Two close contacts became symptomatic and required evaluation. Challenging process issues included tracing of laboratory specimens, decontamination of laboratory equipment, and internal and external communication. After 5 days, a transmissible VHF of public health consequence was ruled out in the index case. CONCLUSION: Contingency plans for VHF can be implemented in an efficient and feasible manner. Contact tracing, laboratory issues, internal communication, and media interest can be anticipated to be the key challenges. PMID- 12725357 TI - The epidemiology of multidrug-resistant Acinetobacter baumannii: does the community represent a reservoir? AB - OBJECTIVE: To explore the role of the community as a potential reservoir for Acinetobacter baumannii. DESIGN: Antimicrobial resistance patterns and genotypes of A. baumannii isolates from patients in two Manhattan hospitals were compared with those of A. baumannii isolates from the hands of community members. RESULTS: A total of 103 isolates from two hospitals (hospital A, 81; hospital B, 22) and 23 isolates from community residents were studied. Of the hospital isolates, 36.6% were multidrug resistant (hospital A, 68.2%; hospital B, 27.8%). In contrast, there were no multidrug-resistant isolates from the community (P < .005 between hospital and community). The prevalence of A. baumannii on the hands of community residents was 10.4% (23 of 222). By molecular typing, 42 strains of A. baumannii were identified. Of the isolates from hospital A and hospital B, 55.6% (45 of 81) and 68.2% (15 of 22), respectively, were indistinguishable or closely related. In contrast, most community (83.3%) isolates were unrelated (P = .001 between hospital and community). CONCLUSION: Acinetobacter isolates from the community, characterized by a large variety of unrelated strains (83.3%), were distinct from the hospital isolates, of which 58.3% were closely related. The absence of multidrug-resistant strains in the community compared with 36.8% prevalence among hospital isolates suggests that the reservoir for epidemic strains resides in the hospital environment itself. To our knowledge, this is the first study to examine the community as a potential reservoir for hospital strains of A. baumannii. PMID- 12725360 TI - Infection control in British nursing homes. AB - Infection control in British nursing homes is different from that in U.S. nursing homes in seveal ways. Most British nursing homes, for example, do not have a designated on-site infection control nurse, and several agencies are responsible for regulatory oversight. This article discusses the state of infection control in British nursing homes. PMID- 12725359 TI - Acinetobacter outbreaks, 1977-2000. AB - This review of Acinetobacter outbreaks summarizes factors related to the presence and recognition of organism transmission and describes the implementation of control and prevention measures directed at limiting spread. Exogenous transmission of Acinetobacter should be considered when infections are endemic and when case rates increase. Increasing or new antimicrobial resistances in a collection of isolates also suggest transmission, and transmission can be definitively confirmed when isolates are found to be indistinguishable from or related to one another by a discriminatory genotyping test. An investigation for a common source should be conducted. When a common source cannot be found and eliminated, or once an endemically transmitted organism is established, containment or prevention efforts may require aggressive interventions, complex interventions, or both. Colonization at multiple sites, the relative ease of induction of antibiotic resistance in the organism following patient exposure to multiple drugs, and long-term environmental survival provide enhanced opportunities for the transmission of Acinetobacter between and among patients. New approaches and interventional trials are needed to define effective measures for the prevention and control of Acinobacter infections. PMID- 12725361 TI - Implication of a healthcare worker with chronic skin disease in the transmission of an epidemic strain of methicillin-resistant Staphylococcus aureus in a pediatric intensive care unit. AB - This outbreak of colonization of neonates in a 10-bed pediatric intensive care unit illustrates the probable role of a healthcare worker (HCW) in the transmission of methicillin-resistant Staphylococcus aureus, despite good hygienic practices. It raises the issue of preventive exclusion of HCWs affected by chronic skin disease from high-risk units. PMID- 12725363 TI - Bacterial contamination of computer keyboards in a teaching hospital. AB - We tested 100 keyboards in 29 clinical areas for bacterial contamination. Ninety five were positive for microorganisms. Streptococcus, Clostridium perfringens, Enterococcus (including one vancomycin-resistant Enterococcus), Staphylococcus aureus, fungi, and gram-negative organisms were isolated. Computer equipment must be kept clean so it does not become another vehicle for transmission of pathogens to patients. PMID- 12725362 TI - Mupirocin resistance in clinical isolates of Staphylococcus aureus. AB - One hundred isolates of Staphylococcs aureus were collected in a laboratory serving several hospitals and clinics in southeastern Wisconsin and tested for mupirocin susceptibility. Only two isolates of S. aureus showed mupirocin resistance. The mupirocin-resistant isolates were from hospitalized patients with positive blood cultures. PMID- 12725364 TI - Orthodontic information--a lot more coins in the realm. PMID- 12725365 TI - A 20-year follow-up of signs and symptoms of temporomandibular disorders and malocclusions in subjects with and without orthodontic treatment in childhood. AB - This investigation analyzes the influence of orthodontic treatment on signs and symptoms of temporomandibular disorders (TMDs) and different malocclusions during a 20-year period. Originally, 402 randomly selected 7-, 11-, and 15-year-old subjects were examined clinically and by means of a questionnaire for signs and symptoms of TMDs. The examination was repeated after five and ten years. After 20 years, 320 subjects (85% of the traced subjects) completed the questionnaire. The oldest age group,now 35 years of age, was invited to a clinical examination, and 100 subjects (81% of the traced subjects) were examined. The correlations between signs and symptoms of TMD and different malocclusions were mainly weak, although sometimes statistically significant. Lateral forced bite and unilateral crossbite were correlated with TMD signs and symptoms at the 10- and 20-year follow-ups (r = 0.38, P < .05 and r = 0.34, P < .01, respectively). Subjects with malocclusion over a long period of time tended to report more symptoms of TMD and to show a higher dysfunction index, compared with subjects with no malocclusion at all. There were no statistically significant differences in the prevalence of TMD signs and symptoms between subjects with or without previous experience of orthodontic treatment. This 20-year follow-up supports the opinion that no single occlusal factor is of major importance for the development of TMD, but a lateral forced bite between retruded contact position (RCP) and intercuspal position (ICP), as well as unilateral crossbite, may be a potential risk factor in this respect. Furthermore, subjects with a history of orthodontic treatment do not run a higher risk of developing TMD later in life, compared with subjects with no such experience. PMID- 12725366 TI - Comparison of skeletal and dental morphology in asymptomatic volunteers and symptomatic patients with normal temporomandibular joints. AB - The purpose of this study was to evaluate the skeletal and dental pattern of symptomatic individuals (SN) with normal temporomandibular joints (TMJs). There were 42 symptomatic female patients with bilaterally normal TMJs and 46 asymptomatic normal female volunteers (AV). All study participants had bilateral high-resolution magnetic resonance scans in the sagittal (closed and open) and coronal (closed) planes to evaluate the TMJs. Linear and angular cephalometric measurements were taken to evaluate the skeletal, denture base, and dental characteristics of the two groups. ANOVA was used to compare the symptomatic subjects with the control subjects. There were no significant differences between the two groups besides the lower incisor being more retruded in the asymptomatic group. This study showed that there are no alterations in skeletal morphology in SN individuals compared with AV. PMID- 12725367 TI - Comparison of skeletal and dental morphology in asymptomatic volunteers and symptomatic patients with unilateral disk displacement without reduction. AB - The purpose of this study was to evaluate the effect of unilateral disk displacement without reduction (UDDN) on the skeletal and dental pattern of affected individuals. There were 12 symptomatic female patients and 46 asymptomatic normal female volunteers. All study participants had bilateral high resolution magnetic resonance scans in the sagittal (closed and open) and coronal (closed) planes to evaluate the temporomandibular joints. Linear and angular cephalometric measurements were taken to evaluate the skeletal, denture base, and dental characteristics of the two groups. ANOVA was used to compare the symptomatic subjects with the control subjects. A few skeletal differences were found. There was an overall reduction in length of the anterior (S-Na) and total (S-Ba) cranial base measurements in the UDDN group. The mandibular plane angle was steeper and the posterior ramal height (Ar-Go) was shorter in the symptomatic group. The only dental difference found was a relative infraeruption of the lower first molar. This study suggests that subjects with UDDN may manifest altered craniofacial morphology. Although the cephalometric measurements used did not account for any asymmetry, previous studies have shown that UDD may cause mandibular asymmetry. Presence of asymmetry and altered craniofacial morphology should alert the clinician especially while orthodontically treating children and surgical patients. PMID- 12725368 TI - Biomechanics of craniofacial sutures: orthopedic implications. AB - Sutures are soft connective tissue articulations between craniofacial bones. Suture mechanics deals with patterns of mechanical stress experienced in sutures resulting from natural activities such as mastication and exogenous forces such as orthopedic loading. Patterns of sutural mechanical stress can be delineated readily as sutural strain using strain gages attached over the suture. In mastication, complex sutural strain patterns have been elucidated in a few species. Mechanical stresses are not transmitted in the skull as a continuing gradient, for different sutures are capable of redefining a propagating mechanical force as predominately tensile or compressive strain. Exogenous mechanical forces with engineering waveforms such as static and sine wave at different frequencies induce corresponding waveforms and rates of sutural strain, providing the basis for applying novel mechanical stimuli to engineer sutural growth. The available data on suture mechanics converge to a hypothetical theme that mechanical forces regulate sutural growth by inducing sutural mechanical strain. Various orthopedic therapies, including headgear, facemask, and functional appliances may induce sutural strain, leading to modification of otherwise natural suture growth. PMID- 12725369 TI - Children with class III malocclusion: development of multivariate statistical models to predict future need for orthognathic surgery. AB - Until now, the literature does not provide an accurate model to predict the future need for orthognathic surgery in prepubertal patients with class III malocclusion. Because not all of these patients are candidates for later surgical correction, patient assessment and selection remain arbitrary with respect to diagnosis and treatment planning. The purpose of the present investigation was to analyze the value of classifying class III children before puberty into patients who can be effectively treated by orthopedic/orthodontic therapy alone and those who require orthognathic surgery. To obtain a robust model, the study design was multicentric (University Orthodontic Departments of Frankfurt, Heidelberg, and Wurzburg). A total of 88 patients with class III malocclusion were grouped into orthopedic/orthodontic (n = 65) and surgery patients (n = 23), according to their records after puberty (mean age, 17 years three months). Discriminant analysis (DA) and logistic regression (LogR) were applied to 20 landmarks of the patients' cephalograms before puberty (mean age, nine years eight months) to identify the dentoskeletal variables that provide the best group separation and the best predictability of group membership, respectively. Both models were highly significant (P < .001), classifying 93.3% (DA) and 94.3% (LogR) of the patients correctly. The extracted variables were identical for both procedures: Wits appraisal, palatal plane angle, and individualized inclination of the lower incisors. The resulting equation of LogR was individual score = -7.968 - 1.323Wits - 0.363NL-NSL + 0.153[180 - (LI-ML) - (L1-ML(ind))]. We concluded that by means of multivariate statistics, prepubertal children with class III malocclusions may be classified into nonsurgery and surgery patients with high accuracy. PMID- 12725370 TI - Vertical growth changes after adenoidectomy. AB - The purpose of this retrospective investigation is to compare vertical growth component of craniofacial structure of subjects with early and late adenoidectomy history. The study consisted of 93 lateral cephalometric radiographs of three groups of randomly selected patients. The first group was made up of 12 patients (10 male and two female) with an average age of 11.16 +/- 2.08 years, who had been operated upon between 1.5-4 years of age. The second group was made up of 54 patients (25 maleand 29 female) with an average age of 12.18 +/- 2.6 years, who had been operated upon after four years of age. The third group of 27 patients (7 male and 20 female) with clear airway with an average age of 11.18 +/- 2.35 years was used as the control. The data obtained from two adenoidectomy groups were compared and because no statistically significant difference was found except for ANSMe/NMe, the two groups were pooled and compared with the growth pattern in the control sample. There were statistically significant differences in the following parameters: SNGoMe, PPGoMe angle, Gonial angle, Gonial ratio, sigma of inner angles, ANSMe/NMe ratio, Jarabak ratio, PNS-adl distance, PNS-ad2 distance, OAW1 distance. When compared with the control group, the adenoidectomy group showed a more vertically directed growth pattern, however, there were no vertical growth pattern differences between the two groups of children who had adenoidectomy before and after four years of age. PMID- 12725371 TI - Effects of experimental nasal obstruction on human masseter and suprahyoid muscle activities during sleep. AB - The effect of nasal obstruction on nocturnal masseter and suprahyoid muscle activities using a newly developed portable electromygram (EMG)-recording unit was examined. Ten healthy Japanese males participated in this study. EMG activities of both the right masseter and bilateral suprahyoid muscles were recorded with a portable EMG-recording unit. At midnight, the subject was asked to lie on a bed after complete preparation with surface electrodes. After maximal clenching and jaw-opening effort (100% maximum voluntary contribution), the subject was allowed to fall asleep. In the first half of the night, EMG activities were recorded for about three hours of sleep without nasal obstruction. In the second half of the night, EMG activities were recorded for about three hours of sleep with nasal obstruction. In both muscles, there were no significant changes in either the maximal EMG activities or the number of events beyond 40% MVC with nasal obstruction. But in an evaluation based on the distribution of muscle activities, the EMG activity of the masseter muscle tended to decrease (P = .07) and that of the suprahyoid muscles increased significantly (P = .02) with nasal obstruction. These results suggest that nasal obstruction could modulate the activities of the masseter and suprahyoid muscles during sleep. PMID- 12725372 TI - Effects of maxillary molar intrusion on the nasal floor and tooth root using the skeletal anchorage system in dogs. AB - The skeletal anchorage system (SAS) was developed to provide intraoral absolute anchorage for the intrusion or distalization of molars. The purpose of this study was to verify the effects of remarkable molar intrusion on the tooth root and the maxillary sinus floor. Six adult female beagles with fully erupted dentition were used. Titanium miniplates were implanted bilaterally above the maxillary second premolar root apices using pentobarbital anesthesia. The second premolars were intruded for four or seven months after three months of healing after implantation. Standardized dental radiographs were taken periodically to evaluate the amount of tooth movement and root resorption. After the experimental animals were fixed by perfusion at the end of each experimental period, the second premolars were dissected along with the surrounding alveolar bone. Undecalcified (60 microm thick) and decalcified (five microm thick) sections were prepared. The average extent of intrusion was 1.8 mm after four months and 4.2 mm after seven months. The root apices of the intruded molars penetrated into the nasal cavity. Remodeled bone around the intruded molar toots was rich in woven bone on the buccal side, whereas that on the palatal side was rich in lamellar bone. Nasal floor membrane and a thin layer of newly formed bone, which lifted intranasally, covered the intruded molar root. Root resorption partly reached into the dentine without the formation of reparative cementum, and little or no serious pathological changes were seen in the pulp of the intruded molars. SAS effectively intruded maxillary molars, but some moderate root resorption was observed. PMID- 12725373 TI - Influence of stainless steel inserts on the resistance to sliding of esthetic brackets with second-order angulation in the dry and wet states. AB - Stainless steel (SS) inserts have been added to plastic, ceramic, and composite brackets to improve their frictional characteristics while preserving their esthetic appeal. When coupled with SS archwires, the resistances to sliding of esthetic brackets with and without SS inserts were compared with control brackets of SS. The resistances to sliding were measured in both the dry and wet (saliva) states at 32 second-order angles between -12 and + 12 degrees. When clearances existed between the walls of the brackets and the archwires, the resistances to sliding for the esthetic brackets without inserts were between 38 cN in the dry state and 73 cN in the wet state; those of the esthetic brackets with inserts ranged from 42 cN in the dry state to 65 cN in both states. The resistances to sliding of the SS brackets equaled 38 and 52 cN in the dry and wet states, respectively. When clearances no longer existed, the resistances to sliding for the esthetic brackets with and without inserts generally increased with angulation at a rate equal to or greater than that of the SS brackets--except for the polycarbonate (PC) brackets in the dry state. Because PC brackets without inserts elastically deformed, they had lower resistances to sliding when deformation occurred. For the polycrystalline alumina brackets without inserts, the resistances to sliding increased rapidly and nonlinearly as angulation increased above 4.8 degrees. Upon examination, the presence of scratches on the archwires and SS debris on the brackets was observed. The addition of these particular SS inserts did not considerably improve the resistance to sliding over those esthetic brackets without inserts. PMID- 12725374 TI - Bond strength with custom base indirect bonding techniques. AB - Different types of adhesives for indirect bonding techniques have been introduced recently. But there is limited information regarding bond strength with these new materials. In this in vitro investigation, stainless steel brackets were bonded to 100 permanent bovine incisors using the Thomas technique, the modified Thomas technique, and light-cured direct bonding for a control group. The following five groups of 20 teeth each were formed: (1) modified Thomas technique with thermally cured base composite (Therma Cure) and chemically cured sealant (Maximum Cure), (2) Thomas technique with thermally cured base composite (Therma Cure) and chemically cured sealant (Custom I Q), (3) Thomas technique with light-cured base composite (Transbond XT) and chemically cured sealant (Sondhi Rapid Set), (4) modified Thomas technique with chemically cured base adhesive (Phase II) and chemically cured sealant (Maximum Cure), and (5) control group directly bonded with light-cured adhesive (Transbond XT). Mean bond strengths in groups 3, 4, and 5 were 14.99 +/- 2.85, 15.41 +/- 3.21, and 13.88 +/- 2.33 MPa, respectively, and these groups were not significantly different from each other. Groups 1 (mean bond strength 7.28 +/- 4.88 MPa) and 2 (mean bond strength 7.07 +/- 4.11 MPa) showed significantly lower bond strengths than groups 3, 4, and 5 and a higher probability of bond failure. Both the original (group 2) and the modified (group 1) Thomas technique were able to achieve bond strengths comparable to the light cured direct bonded control group. PMID- 12725375 TI - A comparison of dynamic and static testing of latex and nonlatex orthodontic elastics. AB - The purpose of this study was to determine the effects of repeated stretching (cyclic testing) and static testing on the force decay properties of two different types of orthodontic elastics from a single supplier. Samples of American Orthodontics' 0.25 inch, 4.5 oz (6.35 mm, 127.5 g) latex and nonlatex elastics were used and a sample size of 12 elastics per group was tested. Static testing involved stretching the elastics to three times marketed internal diameter (19.05 mm) and measuring force levels at intervals over 24 hours. Cyclic testing used the same initial extension but cycled the elastics an additional 24.7 mm to simulate extension with maximal opening in the mouth. Both types of elastic had similar initial forces that were statistically below the marketed force (122 and 118 g for latex and nonlatex elastics, respectively) at three times marketed internal diameter. Cyclic testing caused significantly more force loss and this difference occurred primarily within the first 30 minutes. For statically tested elastics the percentage of initial force remaining at 4, 8, and 24 hours was 87%, 85%, 83%, and 83%, 78%, 69% for latex and nonlatex elastics, respectively. For cyclically tested elastics the percentage of initial force remaining at 4, 8, and 24 hours was 77%, 76%, 75%, and 65%, 63%, 53% for latex and nonlatex elastics, respectively. PMID- 12725376 TI - Longitudinal study of anteroposterior and vertical maxillary changes in skeletal class II patients treated with Kloehn cervical headgear. AB - This is a study to evaluate the posttreatment and long-term anteroposterior and vertical maxillary changes in skeletal Class II Division 1 patients (ANB > or = 5 degrees) who had received Kloehn cervical headgear treatment. The sample consisted of 120 lateral cephalograms obtained at pretreatment (T1), posttreatment (T2), and postretention (T3) phases of 40 patients (18 males and 22 females). The patients were of an average age of 10% years in phase T1, 13% years in phase T2, and 23% years in phase T3. They were treated with cervical traction and an expanded inner bow (4-8 mm) and a long outer bow bent upwards off the horizontal 10-20 degrees in relation to the inner bow. After correction of the molar relationship on both sides, a conventional edgewise fixed appliance was used to complement the correction of the malocclusion. The onset of treatment was either at the late mixed dentition or at the beginning of the permanent dentition. The force applied for the 40 patients averaged 450 g and the recommended use of the appliance was 12-14 hours per day with monthly adjustments. F-Snedecor test was applied to the entire sample and multiple comparisons between phases were tested by the Bonferroni method. Results revealed that treatment had reduced maxillary protrusion, inclined the palatal plane with an increase in the SN-PP angle with reduction at long-term. In conclusion, Kloehn cervical headgear with elevated external bow and expanded inner bow was efficient in correcting the skeletal Class II in late mixed-early permanent dentition. Skeletal Class II correction with Kloehn cervical headgear was found to be very stable long term. PMID- 12725377 TI - Mandibular symphyseal distraction and its geometrical evaluation: report of a case. AB - In this report, the case of a patient who has been treated with a different use of a tooth-borne custom-made mandibular symphyseal distraction device is presented. The difference in the application is that the distal arm of the device was sectioned during the retention phase to allow the possible relapse of displaced condyles to their original positions while the labial segment expansion is being maintained. The effect of this procedure was also evaluated on a geometrical model using measurements from the patient's cast. We conclude that symphyseal distraction is an effective and fast method of correcting orthodontic anomalies. The effect of the procedure on the condyle was only 3 degrees of distolateral rotation as calculated using the geometrical model. PMID- 12725378 TI - Maxillary anterior segmental advancement by using distraction osteogenesis: a case report. AB - After the first clinical application of distraction osteogenesis (DO) to correct mandibular deformity was reported in 1992, various applications such as maxillary or midface advancement, temporomandibular joint reconstruction, alveolar augmentation, and mandibular widening have been described in the oral and maxillofacial region. Block et al and Altuna et al first examined anterior segmental DO experimentally in the maxilla and reported successful results. After these studies, DO has been used clinically for the total advancement of the maxilla or midface. But no clinical application of DO for maxillary anterior segmental advancement was found by a review of the literature in English. In this article, we present a case with a skeletal Class III abnormality resulting from a maxillary deficiency, which was treated by using anterior segmental DO. PMID- 12725379 TI - Severe anterior open bite malocclusion with multiple odontoma treated by C lingual retractor and horseshoe mechanics. AB - A fixed lever arm appliance called C-lingual retractor was placed on the lingual aspects of the maxillary anterior teeth in a 16-year-old male patient with a Class II anterior open bite malocclusion. The treatment plan consisted of extracting both upper first premolars and retracting the upper six anterior teeth. A multiple odontoma between the lower left lateral incisor and canine was surgically removed before orthodontic treatment, and a horseshoe appliance was used in the lower dentition for intermaxillary anchorage during the bone-healing period. The transpalatal arches soldered to the upper first and second molar bands were used as an intra-arch anchor unit for upper-space closure. Class II elastics were used buccally between the upper six anterior teeth and the lower horseshoe appliance. We took 13 months to treat the open bite malocclusion. There was a decrease in lip fullness as the upper anterior teeth were retracted, which contributed to a decrease in facial convexity. The treatment result was maintained six months after debonding. Details of the new appliance, clinical procedures, and treatment changes are presented. PMID- 12725380 TI - Good preclinical bioanalytical chemistry requires proper sampling from laboratory animals: automation of blood and microdialysis sampling improves the productivity of LC/MSMS. AB - The preclinical bioanalytical process with animal models begins with sampling biological fluids and tissue. The goal is to understand oral absorption kinetics, distribution, metabolism, excretion, blood brain barrier penetration, drug-drug interactions, and the influences on biomarkers, hematology, electrophysiology, cardiology, blood pressure and behavior. An overview is obtained by periodic blood sampling of 8-12 samples over a total time span of 10-24 h. Urine, feces, bile and microdialysates can augment the information available from whole blood. In today's preclinical environment, the majority of samples are processed by LC/MSMS augmented by robotic sample preparation tools. These tools save labor and improve precision for smaller volume/lower concentration samples. Our laboratories have been engaged in a project that is focused on improving both the quality and throughput for laboratory animal studies, while providing for reduced numbers of animals and enhanced animal comfort. We have implemented a robotic system that can accomplish most of the above goals for laboratory rats, dogs and primates. Studies with mice are at an earlier stage, but feasibility has been demonstrated. This presentation is a progress report on this evolving research program in cooperation with multiple pharmaceutical and drug development companies. We will illustrate results and discuss future directions. PMID- 12725381 TI - Surface-analytical studies on environmental and geochemical surface processes. AB - The surface chemical compositions of solid samples from environmental and geological sources can differ from the bulk or average compositions, because of changes caused by adsorption, dissolution, oxidation, etc. Accordingly, analytical information on surface layers is important for a better understanding of the environmental chemistry involving solid surfaces. The rapid development of surface-analytical techniques has enabled us to probe the surface chemistry of environmental and geological solid samples of complex composition. This article demonstrates some examples of important items of information that can be obtained by applying surface-analytical techniques, such as X-ray photoelectron spectroscopy and secondary ion mass spectrometry, to environmental and geological samples. The surface analysis of fly ashes, soils, sediments and weathered silicate minerals is reviewed. PMID- 12725382 TI - A screening method for estrogens using an array-type DNA glass slide. AB - A new screening assay was described for the determination of endocrine disrupting chemicals (EDCs), such as synthetic estrogens, with an array-type DNA glass slide having characteristics of 1) a high sample throughput, 2) a compact size allowing a small sample volume, and 3) a sensitive determination based on the estrogen dependent binding of the human estrogen receptor a (hERalpha) with its estrogen responsive element (ERE; Vit. A2 gene promoter). We devised a glass slide on which a thin agarose gel was mounted. Avidin was then covalently immobilized on each well of the glass slide after the gel was activated by a NaIO4 solution. Also, the biotinylated ERE as a DNA probe was immobilized on the gel layer through avidin-biotin binding. After the estrogen-dependent binding of a yellow fluorescent protein-fused hERalpha (YFP-hERalpha) to ERE on the gel layer, the fluorescence intensity of YFP-hERalpha quantitatively extracted into the gel was directly determined with a fluorescence microplate reader. Pre-incubation of YFP hERalpha with estrogen at 37 degrees C for 30 min enhanced the estrogen-dependent hERalpha-ERE binding. The determined hormonal activities of estrogens on the interaction of YFP-hERalpha with ERE were as follows in their decreasing order: diethylstilbestrol (DES) > 17beta-estradiol (E2) <==> ethynylestradiol (EE2) > 4 hydroxy tamoxifen (OHT) > clomiphene (Clo). The present method provides a sensitive estrogen-dependent dose-response curve down to approximately 10(-13) M in the case of DES. This method will become a competitive alternative to the conventional in vitro assays, such as a DNA-binding assay using radioisotopes. PMID- 12725383 TI - Monitoring of oxidation steps of ascorbic acid redox reaction by kinetics sensitive voltcoulometry in unsupported and supported aqueous solutions and real samples. AB - Aqueous solutions of ascorbic acid in unsupported and supported aqueous solutions and real samples were studied by the kinetics-sensitive double-step voltcoulommetric method with the aim to contribute to a better understanding of its behavior in biological systems. The data obtained from measurements made on analytes prepared in the laboratory, as well as those made on real samples (some commercial orange drinks, flash of the fresh fruits) point to the redox reaction of L-ascorbic acid (L-AH2) being very sensitive to both the presence of dissolved gaseous species (O2, CO2) and the ionic strenght in the analyte. Either the dissolved gaseous species, or the higher ionic strength caused by both the presence of supporting electrolyte and increased total concentration of ascorbic acid, respectively, give birth to the degradation of L-AH2. Naturally, the highest percentage of L-AH2 was spotted in fresh fruit. PMID- 12725384 TI - Differential pulse adsorptive stripping voltammetric determination of dinoseb and dinoterb at a modified electrode. AB - A sensitive adsorptive stripping voltammetric method for the determination of dinitrophenolic herbicides, dinoseb (DSB) and dinoterb (DTB) at a bare carbon paste electrode (CPE) and a clay modified carbon paste electrode (CMCPE) was developed. A systematic study of various experimental conditions, such as the pH, accumulation variables and composition of a modifier on the adsorptive stripping response, were examined by using differential pulse voltammetry. A significant improvement was observed in the sensitivity by using the present method with CMCPE. When CMCPE was used, a linear response was obtained over the concentration range 2 x 10(-10) to 3 x 10(-7) M and 6 x 10(-10) to 6 x 10(-7) M with lower detection limits of 1 x 10(-10) M and 5.4 x 10(-10) M for dinoseb and dinoterb, respectively, at an accumulation time of 100 s. The interference from other herbicides and ions on the stripping signals of both compounds was also evaluated. The described method was applied to estimate of the dinoseb and dinoterb in environmental samples. PMID- 12725385 TI - Selective measurement of gaseous hydrogen peroxide with light emitting diode based liquid-core waveguide absorbance detector. AB - Atmospheric H2O2 is typically determined by enzymatically mediated fluorogenic reactions that do not discriminate between H2O2 and organic peroxides. Reactions of Ti(IV) with H2O2 has also been the basis of colorimetric measurements of H2O2 but is too insensitive. A more sensitive determination is possible with the Ti(IV)-4-(2-pyridylazo)resorcinol (PAR) complex, however, unreacted PAR must be chromatographically separated. A titanium(IV)porphyrin complex, oxo[5,10,15,20 tetra(4-pyridyl)porphyrinato]titanium(IV) [TiO(tpypH4)4+], (TiTPyP) was introduced for the measurement of aqueous H2O2. In this paper, we show that TiTPyP can be used for measuring H2O2(g)), it does not respond to CH3HO2. With a proper membrane collector, practically there is no interference from concurrently present gaseous SO2 and O3. The approach permits a S/N = 3 limit of detection (LOD) of 26 pptv with a 50 mm path liquid core waveguide (LCW) absorbance detector and a light emitting diode based light source. This is adequate for real atmospheric measurements. PMID- 12725386 TI - Determination of elemental and ionic compositions for diesel exhaust particles by particle induced X-ray emission and ion chromatography analysis. AB - The purpose of this study is to clarify the chemical characterization of PM2.5 and PM10 in diesel exhaust particles (DEP). Sampling of PM2.5 and PM10 in DEP was carried out in November 1999 using an automobile exhaust testing system at the National Traffic Safety and Environment Laboratory, with a diesel truck (engine type: direct injection, displacement: 7,961 cc, carrying weight: 2,020 kg, equivalent inertia weight: 5,600 kg) placed on a chassis dynamometer. Sampling conditions included idling, constant speed of 40 km/h, M-15 test pattern and 60% revolution/40%-load of maximum power. Samples were collected on a polycarbonate membrane filter (Nuclepore, pore size: 0.8 microm) using a MiniVol Portable Air Sampler (Airmetrics Co., Inc.). The concentrations of several elemental and ionic species in the PM2.5 and PM10 samples were determined by particle induced X-ray emission (PIXE) and ion chromatography analysis. PIXE analysis of the PM2.5 and PM10 samples revealed 15 elements, of which Na, Mg, Si, S, Cl, Ca, Fe and Zn were found to be the major components. Ionic species were Cl-, NO2-, NO3-, SO4(2-), Na+, NH4+, K+ and Ca2+. Concentrations of elements and ionic species under the sampling condition of 60%-revolution/40%-load were highest in comparison with those of the other sampling conditions. The elemental and ionic species data were compared for PM2.5 and PM10; PM2.5 concentrations were 70% or more of PM10 concentrations for the majority of elements, and concentrations of ionic species in PM2.5 and PM10 were almost identical. PMID- 12725387 TI - Determination of trace metal complexes by natural organic and inorganic ligands in coastal seawater. AB - It is well-documented that organic compounds form strong complexes with most metals in aquatic systems, and that seawater is a complex medium which contains a large variety of organic and inorganic ligands, including colloidal matter. We suggest that most trace metals are complexed in seawater and that some inorganic metals complexes are either labile or not stable. In contrast, metal-organic complexes are often stable and need various and specific treatments to be dissociated. In this paper we try to illustrate a good tendency of some trace metals to be complexed by organic ligands in seawater. A solid-phase extraction method was applied using a C18 column as a resin that is able to separate metals complexed by neutral organic ligands, and the chelamine resin to separate metal species that are present as labile inorganic complexes. The determination of total dissolved metal concentrations was achieved by formatting a metal-8 hydroxyquinoline complex, followed by adsorption on C18 columns and ICP-AES analysis. PMID- 12725388 TI - Identification of electrical degradation products of 4-chlorophenol in water. AB - The electrical decomposition of 4-chlorophenol in water was examined with iridium dioxide doped on atitanium electrode. A number of electrical degradation products of 4-chlorophenol, such as hydroquinone and chlorohydroquinone via the addition of hydroxyl radicals, and dichlorophenol through addition of chlorine radical, were observed as major products. Moreover, hydroxylated chlorobiphenylethers, hydroxylated dibenzo-p-dioxin/furans and hydroxylated chlorobiphenyls formed by a dimerization process during the electrolysis process of 4-chlorophenol were also observed. On the other hand, benzoquinone, muconic acid and aldehyde derivatives that were further oxidative products of hydroquinone formed by photocatalysis process, were not observed. The electrical decomposition products of 4 chlorophenol were trimethylsilylated and then identified by gas chromatography mass spectrometry. The degradation rate of 4-chlorophenol in water by iridium oxide electrode was measured against the electrical process duration. After iridium electrical process for 120 min, about 50% of 4-chlorophenol was converted into a number of products through oxidation processes. On the basis of the identified products, the degradation pathways of 4-chlorophenol under electrolysis process were proposed. PMID- 12725389 TI - Simultaneous analysis of carbamate pesticides in tap and raw water by LC/ESI/MS. AB - Ten carbamate pesticides including four suspected endocrine disruptors, methomyl, benomyl (carbendazim), aldicarb and carbaryl, were simultaneously analyzed by LC/ESI/MS. The influence of the matrix on the variation of the ion signal intensities of (M + H)+ and adduct ions was investigated. Although the intensities of three oxamyl ions changed depending on the matrix, the variation in the concentration calculation of oxamyl was reduced by using the sum total of the area value of two ions. The limits of the quantitation of ten pesticides without a concentration procedure were from 0.4 - 30 microg/l. The solid-phase recovery rates of ten pesticides spiked into tap water and raw water were in the range of 69-111%. Using this method, the concentrations of the pesticides in tap and raw water sampled at 14 monitoring points in Hyogo Prefecture were determined. Carbendazim in three raw water samples and carbofuran in one of these three samples were detected at low concentrations (less than 0.32 microg/l). PMID- 12725390 TI - Aquacyanocobalt(III)-cobyrinate as a key compound in NO2-sensitive polymeric liquid membranes. AB - The reactions within a recently introduced NO2-sensitive polymeric membrane based on aquacyanocobalt(III)-cobyrinate are described. The detailed reaction mechanism was investigated in three ways: using UV/VIS-spectroscopy, determining the reaction products and investigating the influence of other gases. It could be shown that the membrane's high sensitivity and selectivity derives from the remarkable reaction of NO2 with this cobyrinate derivative, which yields nitric and nitrous acid. This reaction can be transduced with a consecutive protonation of the included chromoionophore into an optical signal, which allows the detection of NO2 in the ppb-range. PMID- 12725391 TI - Simultaneous determination of copper(II) and cobalt(II) by ion chromatography coupled with chemiluminescent detection. AB - In the absence of any special luminescent reagents, a weakly chemiluminescent emission was observed during the decomposition of hydrogen peroxide, catalyzed by transition-metal ions, such as Cu(II) and Co(II), in basic aqueous solution. The chemiluminescent intensity was significantly enhanced by the addition of ethyldimethylcetylammonium bromide and uranine. The signal-to-noise ratio (S/N) was proportional to the concentrations of Cu(II) and Co(II). Based on these phenomena, a flow-injection chemiluminescent method for the simultaneous separation and determination of Cu(II) and Co(II) was developed. The detection limits of the present chemiluminescent method for Cu(II) and Co(II) were 7.5 and 0.01 ng/ml, respectively. After ion chromatographic separation of Cu(II) and Co(II) by an IonPac CS5A column with oxalic acid and lithium hydroxide monohydrate as the eluent, the present chemiluminescent system was used as a post column detector for these two transition metal ions in natural water samples. PMID- 12725392 TI - Determination of butamyrate citrate in cough preparations by derivative UV spectrophotometry and high performance liquid chromatography. AB - Derivative spectrophotometric procedures and an isocratic high performance liquid chromatographic method for the determination of butamyrate citrate (Sinecod, Safarol) in cough syrups have been developed. In the spectrophotometric method, direct measurement of the drug at its absorption maxima is impossible because of interference from different absorbing excipients. Extraction of butamyrate citrate was performed with n-pentane/isopropyl alcohol. Quantification was carried out through the use of 1D derivative at a trough depth of 253.6 nm where interferences from other coextracted compounds are negligible. The extraction efficiency expressed as a % recovery and precision were assessed by fortifying placebo syrup(s) with known amounts of the compound. Also, a reversed phase high performance liquid chromatographic method was used with a mobile phase containing 0.015 M aqueous tetraethylammonium hydrogen sulfate, methanol and acetonitrile 40:30:30 adjusted to pH 3.50 with ammonium hydroxide. The retention behavior of butamyrate citrate as a function of both pH and salt concentration in the aqueous portion of the mobile phase was investigated. Quantification was achieved with UV detection at 258 nm based on peak area. The HPLC method clearly separates the analyte from its degradation products derived after storage of samples under different stress conditions such as acid, alkaline, temperature, oxygen and light. The described methods were successfully applied to the determination of butamyrate citrate in commercial pharmaceutical products and in placebo syrups prepared in the laboratory with good accuracy and precision. The results of the present study show that the use of the derivatives and the HPLC procedure provide precise and sensitive methods for the determination of the compound in pharmaceutical formulations. PMID- 12725393 TI - Flow injection analysis of hydrogen peroxide using glass-beads modified with manganese(III)-tetra(4-carboxyphenyl)porphine derivative and its analytical application to the determination of serum glucose. AB - A flow injection analysis system of hydrogen peroxide was developed. The present system is based on measuring of the absorbance of a quinoid dye formed by the following reaction catalyzed by peroxidase: Phenol + 4-Aminoantipyrine + 2H2O2 - > Peroxidase --> Quinoid dye + 4H2O. A column packed with aminopropyl-glass beads modified with amanganese(III)-tetra(4-carboxyphenyl)porphine derivative (Mn TCPP(G) column), which has peroxidase-like activity, was used in place of an immobilized peroxidase column in the above reaction. The linear range of the calibration curve was 0.4-80 microg/ml hydrogen peroxide. The relative standard deviation of this system was 2.97% (n = 100, 10 microg/ml hydrogen peroxide 20 microl injection). The Mn-TCPP(G) column has sufficiently stability for the continuous injection of hydrogen peroxide untill 100 times. The advantageous feature of the Mn-TCPP(G) column was a less-electrostatic interaction between the mother glass beads and the anionic chromogen or quinoid dye formed and the stability in terms of the storage, temperature and moisture. The determination of serum glucose was achieved by attaching an immobilized glucose oxidase column to this system without deproteinization. PMID- 12725394 TI - Electrochemiluminescent determination of L-cysteine with a flow-injection analysis system. AB - A flow-injection electrochemiluminescent method for L-cysteine determination has been developed based on its enhancement of the electrochemiluminecence of luminol at a glassy carbon electrode. This method is simple and sensitive for cysteine determination. Under the selected experimental parameters, the linear range for cysteine concentration was 1.0 x 10(-6) - 5.0 x 10(-5) mol/l, and the detection limit was 0.67 micromol/l (S/N = 3). The relative standard deviation for 11 measurements of 1.0 x 10(-5) mol/l cysteine was 4.5%. The proposed method has been applied to the detection of cysteine in pharmaceutical injections with satisfactory results. PMID- 12725396 TI - Determination of lead in seawater by flow-injection on-line preconcentration electrothermal atomic absorption spectrometry after coprecipitation with iron(III) hydroxide. AB - A flow-injection on-line preconcentration-electrothermal atomic absorption spectrometric (ETAAS) method coupled with a coprecipitation method has been developed for the determination of lead in seawater. The combination of two preconcentration procedures, coprecipitation with iron(II) hydroxide and solid phase extraction with a lead-selective resin, Pb-Spec, allowed the determination of lead at the ng kg(-1) level. Lead in 250 g of a sample solution was collected by coprecipitation with 10 mg of iron. The precipitate was dissolved in 25 ml of 1 mol l(-1) nitric acid; then, a 4-ml aliquot of the sample solution was introduced into the flow-injection system to preconcentrate and separate lead from iron on a Pb.Spec microcolumn. The sorbed lead was eluted with a 1.0 x 10( 4) mol l(-1) EDTA solution. The 30-microl portion of the eluate corresponding to the highest analyte concentration zone was injected into a graphite furnace. The overall enhancement factor was about 200 for 250 g of the sample. The average and standard deviation of ten blank values obtained were 1.7 ng and 0.38 ng, respectively. The recovery was 93.7 +/- 5.0% for seawater spiked with 20 ng kg( 1) lead. The proposed method is applicable to the analysis of seawater for lead at slightly higher levels. PMID- 12725395 TI - Sensitive and simple cloud-point preconcentration atomic absorption spectrometry: application to the determination of cobalt in urine samples. AB - A new approach for developing a cloud-point extraction-flame atomic absorption spectrometric method has been described and used for the determination of cobalt. In this approach, water was removed from the final diluted surfactant rich phase obtained in cloud-point extraction procedure. The results indicated that removing water from this phase increased the enhancement factor by 4-fold. 1-(2 Pyridylazo)-2-naphthol (PAN) and octylphenoxypolyethoxyethanol (Triton X-114) were used as a hydrophobic ligand and a nonionic surfactant, respectively. The chemical variables affecting the preconcentration step were optimized. The effect of the water concentration in the final diluted methanolic surfactant solution on the analytical signal was investigated. The results showed that the analytical signal decreased by 30% and 52% in 15% and 25% water concentrations in methanol, respectively. An enhancement factor of 115 was obtained for cobalt extracted from only 10 ml of a sample. The detection limit obtained under the optimal condition was 0.38 microg l(-1). The proposed procedure was applied to the determination of cobalt in urine samples. PMID- 12725397 TI - Determination of chromium(III) and total chromium in water by derivative atomic absorption spectrometry using flow injection on-line preconcentration with a double microcolumn. AB - A rapid and sensitive method has been proposed for the sequential determination of chromium(III) and total chromium in water samples by flame atomic absorption spectrometry combined with a flow injection on-line preconcentration on a double microcolumn. The chromium(III) and total chromium in samples were retained on a double-microcolumn with a cation exchange resin, respectively, and eluted directly into a nebulizer by 3 mol L(-1) HNO3. The characteristic concentration (gives a derivative absorbance of 0.0044) and the detection limit (3sigma) for chromium were 0.512 microg L(-1) and 0.647 microg L(-1) for a preconcentration time of 1 min, respectively. This is an improvement of 20 and 14-times than those of conventional FI-FAAS. The proposed method allows the determination of chromium in the range of 0-90 microg L(-1) with a relative standard deviation of 3.63% at the 10 microg L(-1) level. The method has been applied for the analysis of chromium in reference water of National Research Center for Certified Reference Materials (GBW08607) and other water samples with satisfactory results. PMID- 12725398 TI - Modified Babington nebulizer for inductively coupled plasma atomic emission spectrometry. AB - A PTFE Babingtonnebulizer equipped with a hood was investigated for inductively coupled plasma atomicemission spectrometry in conjunction with a PTFE cyclone chamber, in order to nebulize various sample solutions containing high salts, hydrofluoric acid and/or suspended solid. A hood of 3 mmphi (nozzle side) - 5 mmphi (outlet side) and 6 mm in length gave a comparable or higher sensitivity compared to a system with a commercially available concentric nebulizer and a glass cyclone chamber. Moreover, the present nebulizer was fully interchangeable with a concentric one at normal argon pressure, attaining sufficient stability, a short wash-out time and good nebulizing of high matrices solutions. The present system was successfully applied to the determination of trace impurities in highly pure silica powders. PMID- 12725399 TI - Photoacoustic determination of iron in corn meal. AB - We report here on the use of the photoacoustic technique for the determination of low concentrations of total iron in corn meal samples. The determination of this element in food is of considerable interest because several foods are currently enriched with it at proper levels in order to increase the resistance of people, after consumation, to several diseases, such as anemia. The proposed technique is based on an open photoacoustic cell configuration in conjunction with a suitable colorimetric method. We applied it to a measurement of the total iron concentration in corn meal samples. The results agree very well with those obtained using a conventional spectrophotometric method, showing the possibilities of new experimental methodologies based on photothermal methods to perform this kind of study, with the advantage of a higher sensitivity and increment of the range of appreciable absorbance. PMID- 12725400 TI - Distribution of the amplitude of the self-oscillation of the electrical potential difference observed in an organic phase of a water-oil-water liquid membrane system. AB - The electrical potential differences were measured at various points in an organic phase of a water-oil-water liquid membrane system in which self oscillations of an electrical potential difference occurred across a three-phases liquid membrane by using an H-shaped cell. The electrical potential difference oscillated in the direction through the two aqueous phases, but not in its vertical direction. The amplitude of the self-oscillation greatly changed around the place where the horizontal tube was connected to the vertical tube, which had no surfactant. PMID- 12725401 TI - Determination of EDTA species in water by second-derivative square-wave voltammetry using a chitosan-coated glassy carbon electrode. AB - Based on the adsorption of Fe(EDTA)- on a chitosan-coated glassy carbon electrode, a second-derivative square-wave voltammetry for the determination of the EDTA species in water samples was investigated. The measuring range of EDTA was from 6.0 x 10(-7) to 5.0 x 10(-5) mol/L with a correlation coefficient of 0.998 and a detection limit of 2.8 x 10(-7) mol/L. The relative standard deviation was less than 6.2% (n = 5) and the recovery was in the range of 98-105% for the determination of practical samples. The result was consistent with that from the HPLC method. PMID- 12725402 TI - Liquid chromatographic separation and determination of aromatic sulfonates in an aquatic environment using a photodiode array and electrospray ionization-mass spectrometer as detectors. AB - A simple and rapid method involving high-performance liquid chromatographic separation, followed by photodiode array (PDA) and electrospray ionization-mass spectrometric (ESI-MS) detection of aromatic sulfonates in waste-water effluents of industrial units producing optical whitening agents, has been developed. The separation was achieved on a reversed-phase Hypersil C18 column using gradient elution of a mobile phase consisting of 0.05 M ammonium formate-methanol with decreasing concentration of the buffer at room temperature. The minimum detection limits were determined to be in the range of 0.2 - 1.8 x 10(-9) g using PDA and ESI-MS detectors. PMID- 12725403 TI - Preparation of ferrocyanide-imprinted pyridine-carrying microspheres by surface imprinting polymerization. AB - Ferrocyanide-imprinted pyridine-carrying microspheres were prepared using ferrocyanide ions as a template. This method is based on a surface imprinting technique from the seed emulsion that consisted of 4-vinylpyridine (functional monomer), styrene, and butyl acrylate and a water mixture polymerization by a radical initiator. The ferrocyanide-imprinted microspheres had about 200 times higher adsorption affinity over the non-imprinted microspheres in ferrocyanide (template) ion adsorption. This imprinted microspheres also adsorbed other tripolyphosphate, pyrophosphate, and phosphate anions much more strongly than the non-imprinted microspheres did, but were not particularly specific in ferrocyanide ion adsorption. PMID- 12725404 TI - Determination of thiobarbituric acid adduct of malondialdehyde using on-line microdialysis coupled with high-performance liquid chromatography. AB - An on-line analytical system for the continuous monitoring of malondialdehyde (MDA) was developed. This method involves the use of microdialysis perfusion, on line derivatization and on-line HPLC analysis. This method gave a linear response for MDA concentrations and HPLC peak areas in the range from 0.051 microM to 2.43 microM. The intra-day (RSD = 1.6-10.5%) and inter-day (RSD = 1.1-9.3%) precisions were acceptable. The average in vitro probe recovery of MDA standard was 18.4 +/- 1.0%. The detection limit was 0.03 microM, corresponding to 0.6 pmol for an injection volume of 20 microl. This method was used for in vitro peroxidation investigations, which provided evidence for elevated MDA levels following the incubation of metal ions to a linoleic acid solution. PMID- 12725405 TI - Flow-injection determination of ornidazole by chemiluminescence detection based on a luminol-ferricyanide reaction. AB - A flow-injection analysis (FIA) with a chemiluminescence detection method was developed for the determination of ornidazole based on the inhibition intensity of chemiluminescence from the luminol-ferricyanide system. Under the condition of 1.0 x 10(-3) mol/L luminol and 5.0 x 10(-6) mol/L potassium ferricyanide, the response to the concentration of omidazole is linear from 0.2 microg ml(-1) to 10 microg ml(-1), and a detection limit of 0.05 microg ml(-1) can be obtained. This method has been successfully applied to the determination of omidazole in pharmaceutical preparations. PMID- 12725406 TI - Simple method of paroxetine determination using a single channel FIA with no in line reaction process. AB - A flow-injection analysis (FIA) of paroxetine hydrochloride (PRX), a selective serotonin reuptake inhibitor (SSRI) currently used as an antidepressant drug, is described. A 0.1 mol dm(-3) acetate buffer at pH 3.07 was found to be thebest solvent. The analyte was detected at 293 nm. The calibration equation was linear over the range of 1.07 x 10(-6) to 5.35 x 10(-6) mol dm(-3). The limit of detection (LOD) and the limit of quantitation (LOQ) were 3.2 x 10(-7) and 9.5 x 10(-7) mol dm(-3), respectively. The proposed method was applied to the determination of PRX in pharmaceutical preparations. The results were compared with those obtained by a conventional batchwise UV-spectrophotometry. PMID- 12725407 TI - A miniaturized fluorescence detector with a windowless flow cell using a light emitting diode. AB - A miniaturized fluorescence detector using a high-brightness light-emitting diode as an excitation source was constructed and evaluated. A windowless flow cell based on a commercial four-port cross fitting was designed to reduce the stray light level and to eliminate the optical alignment. The observed detection limit for fluorescein was 26 nM in the continuous-flow mode. The error in the reproducibility of the responses was evaluated by the FIA method, and was found to be within 2% RSD. PMID- 12725408 TI - Kinetic-catalytic determination of vanadium(IV) using methyl orange-bromate redox reaction. AB - Determination of V(IV) based on its catalytic effect on the reaction between Methyl Orange and bromate in thepresence of citric acid was studied. The calibration curve obtained by fixed-time method was linear in the range of 2.5 300 ng ml(-1). By use of slope method, a calibration curve containing two linear portions were obtained. Using fixed-time and slope methods, we obtained detection limits of 0.8 and 1.5 ng ml(-1), respectively. Fe2+, As(III), V(V) and Hg2+ interfered. The method was successful for analysis of water samples. PMID- 12725409 TI - Crystal structure and synthesis of 17alpha-(5-chlorovaleroyloxy)-16beta methylpregna-4,6-diene-3,20-dione. AB - C27H37O4Cl is orthorhombic, P2(1)2(1)2(1). The unit-cell dimensions at 293 K are a = 7.1388(15), b = 12.9836(14), c = 26.665(10)A, V = 2471.5(10)A3, Dx = 1.239 g/cm3, and Z = 4. The R value is 0.070 for 1458 observed reflections. The A, B, C and D rings occur in distorted envelope, distorted half-chair, chair, and envelope and a half-chair conformations. The molecules in the crystal are packed at the normal van der Waals distances. PMID- 12725410 TI - Crystal structure of (2,2'-bipyridine-N,N)(2,2',2''-triaminotriethylamine N,N',N'',N''')nickel(II) diperchlorate. AB - An X-ray structure determination shows that the Ni(II) ion is a distorted six coordinated octahedron by four nitrogen atoms of the tetradentate tren ligand constituting the equatorial square base, and by two nitrogen atoms of the bidentate bpy ligand in a cis position. The two six-membered rings of bpy are coplanar and almost pararell. The tetradentate ligand consists of three five membered chelate rings in gauche coformations. The Ni-N(tren) bond lengths of this complex are almost equivalent to the reported values. PMID- 12725411 TI - Crystal structure of (5,14-methyl-7,12-dimethyl-1,4,8,11 tetraazadiiminecyclotetradecylene-N,N',N'',N''')nickel(II) bis(diisopropyldithiophosphate) complex. AB - The macrocyclic tetradentate Ni(II) compound was obtained in a reaction and determined by X-ray diffraction. The crystal structure shows that the molecule is centrosymmetric, and Ni atom located in a square planar coordination environment. The two [(iPrO)2PS2]- anions are outside the macrocycle complex to balance. The Ni-N bond lengths are in the range of 1.906(2) to 1.950(2)A. PMID- 12725412 TI - Crystal structure of [1,4-bis[1-(3,5-dichlorophenolato-2-ylmethyl)-ylpropylamino kappa2N,O]piperazine-kappa2N,N']cobalt(II). AB - The X-ray crystal structure of the title complex, Co(Cl2bprpi) is described. In this complex the Co(II) center displays a distorted octahedral coordination geometry. The piperazine ring exhibits boat conformation, forming chelate rings and capping the Co atom. The N2-Col-N3 angle is 69.41(15) degrees, extremely smaller than 90 degrees. Because the small angle causes a large steric hindrance, the piperazine ring can be effective as an ion size-recognition site. The molecular structure is stabilized by intramolecular N-H...O hydrogen bonds and Cl...H contacts. PMID- 12725413 TI - Crystal structure of 7,8-dihydroxy-4-methylcoumarin. AB - The structure of 7,8-dihydroxy-4-methylcoumarin was determined by an X-ray diffraction method. The compound crystallized in the triclinic space group P1, Z = 2, with a = 7.631(2), b = 9.456(5), c = 7.075(3)A, alpha = 103.13(3), beta = 91.84(3), gamma= 68.21(3) degrees, and V= 460.9(3)A3. The X-ray crystal structure was also compared with those of 5,7-dihydroxy-4-methylcoumarin and 7-hydroxy-4 methylcoumarin. PMID- 12725414 TI - Canine inflammatory mammary carcinoma: histopathology, immunohistochemistry and clinical implications of 21 cases. AB - Human inflammatory breast carcinoma (IBC) is the most malignant type of breast cancer with an extremely poor prognosis. The dog is the unique animal species in which spontaneous inflammatory mammary carcinoma (IC) has been reported, although it is not well documented. The purpose of this study was to characterize histopathologically and immunohistochemically the canine IC, considering associated clinical features. Twenty-one dogs diagnosed with IC and with known clinical and necropsy data were included in the study. Tissue samples from necropsies underwent a histopathological review and an immunohistochemical study (Ki-67, estrogen receptor (ER), progesterone receptor (PR), and P53 tumor suppressor protein). The histological study revealed several types of carcinomas (solid, tubular, papillary, and adenosquamous) and three lipid-rich carcinomas. All tumors were ER negative. Two histological patterns of neoplastic dermal infiltration were observed: tubular/papillary and sarcomatous-like. Dermal sarcomatous-like infiltration was significantly related to previous treatments with progestagens (p = 0.006), primary type of IC (p = 0.03), extreme local pain (p = 0.02), reduced observation of emboli in dermal lymphatic vessels (p = 0.01), and increased expression of p53 (p = 0.001). PR expression was significantly higher in secondary post-surgical IC (p = 0.04). The absence of PR was related to the existence of pulmonary metastases at necropsy (p = 0.04). Canine primary IC is the most aggressive form of this disease with distinct histopathological and immunohistochemical characteristics. Progestins and endocrine-related mechanisms seem to be involved in canine IC development. Canine IC could serve as a spontaneous model for human IBC, particularly in studies concerned with new therapeutics approaches. PMID- 12725415 TI - The prognostic value of the human kallikrein gene 9 (KLK9) in breast cancer. AB - BACKGROUND: Many members of the human kallikrein gene family were found to be differentially expressed in various malignancies and some of them are useful diagnostic/prognostic markers. KLK9 is a newly discovered human kallikrein gene that is expressed in several tissues including thymus, spinal cord, testis, prostate, breast, and ovary. Like other kallikreins, the KLK9 gene was found to be regulated by steroid hormones, mainly estrogens and progestins, in cancer cell lines. EXPERIMENTAL DESIGN: We studied the expression of KLK9 by quantitative RT PCR in 169 breast cancer patients of different stages, grades and histological types. We also compared the relation between KLK9 expression and other clinicopathological variables and patient survival. RESULTS: KLK9 expression is significantly higher in patients with early stage cancers (p = 0.039) and in patients with small tumor size (< 2 cm) (p = 0.028). Kaplan-Meier survival curves demonstrated that KLK9-positive patients have longer disease-free and overall survival (p = 0.015 and 0.036, respectively). Univariate and multivariate analysis also indicates that KLK9 expression is associated with increased disease free and overall survival. When the Cox proportional hazard regression analysis was applied to subgroups of patients, KLK9 expression was found to be a significant predictor of disease-free survival in the estrogen receptor (ER) and progesterone receptor (PR) negative subgroups of patients (Hazard Ratio 'HR' = 0.28, and 0.38, respectively, and p = 0.011 and 0.028, respectively). After adjusting for other known prognostic variables, KLK9 retained its independent prognostic value in these subgroups of patients. Similar results were obtained for overall survival. CONCLUSIONS: KLK9 is a new potential independent marker of favorable prognosis in breast cancer. PMID- 12725416 TI - Breast cell proliferation in postmenopausal women during HRT evaluated through fine needle aspiration cytology. AB - The basis of breast cancer risk associated with hormonal therapies may lie in the regulation of cell proliferation. In a prospective, double-blind, randomized study postmenopausal women were given continuous combined hormone replacement therapy (HRT) either as estradiol valerate 2 mg/dienogest 2 mg, (E2V/DNG) or estradiol 2 mg/noretisterone acetate 1 mg (E2/NETA) for 6 months. Fine needle aspiration (FNA) biopsies were used for immunocytochemical analysis of breast cell proliferation before and during treatment. From 45 women completing the study 135 biopsies were obtained. In the total material there was a more than 4 fold increase in proliferation between baseline and 3 months (p < 0.001). The mean percentage of MIB-1 positive breast cells increased from 2.2 to 9.1%. In some individual women values were as high as 25%. No further increase was recorded at 6 months. While numerical values were somewhat lower in the E2V/DNG group, there were no significant differences between treatments. There was a positive correlation between breast cell proliferation (MIB-1%) and circulating levels of both estradiol (r(s) = 0.54, p < 0.01) and estrone (r(s) = 0.53, p < 0.01) after 3 and 6 months of treatment. No correlations with other endogenous hormones, proteins or with the two exogenous progestogens dienogest and norethisterone were observed. Increased breast cell proliferation should probably be regarded as an unwanted side-effect during HRT. Means to identify those women with the most pronounced proliferative response should be developed. The FNA biopsy technique may be a useful tool to monitor and evaluate the proliferative response to HRT in the normal breasts of postmenopausal women. PMID- 12725417 TI - Estrogen-induced Ets-1 promotes capillary formation in an in vitro tumor angiogenesis model. AB - We employed an in vitro angiogenesis model that simulates the in vivo milieu for tumor capillary formation to study the direct effects of estrogen. 17beta estradiol (E2) treatment significantly stimulated capillary sprouting within 8 h in co-cultures of rat aortic endothelial cells (RAECs) and mouse mammary tumor cells. Co-cultures treated with either progesterone (P4) or E2+P4 showed minimal endothelial cell (EC) sprouting when compared to E2 treated cultures. Treatment with the E2 agonist ICI 182,780 dramatically inhibited capillary formation, demonstrating E2-specificity. Within hours, of E2 treatment ECs isolated from tumor cell/EC co-cultures demonstrated a statistically significant increase in both mRNA and protein levels of the transcription factor Ets-1. We observed increased matrix metalloproteinase (MMP) and decreased tissue inhibitor of metalloproteinase (TIMP) mRNA levels in these ECs following E2 treatment. Ets-1 upregulates expression of the vascular endothelial growth factor (VEGF) receptor, Flt-1 and we detected increased Flt-1 mRNA levels in ECs co-cultured with tumor cells following E2 treatment. Expression of Ets-1 contributes to destabilization of a quiescent EC phenotype in favor of an invasive angiogenic one, in part, by increasing expression of MMPs and integrin molecules that favor migration and invasion. Transfection of ECs with Ets-1 antisense prior to co-culture with E2 resulted in a 95% inhibition in capillary formation. We demonstrate here, for the first time that nanomolar concentrations of E2 directly and rapidly induced new capillary formation in a mammary tumor/EC co-culture system and suggest that this response may be mediated, in part, by an E2-induced increase in Ets-1 expression. PMID- 12725418 TI - The antiproliferative effects of PPARgamma ligands in normal human mammary epithelial cells. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) is a transcription factor in the steroid nuclear receptor superfamily. Ligand activation of PPARgamma is associated with differentiation and an inhibition of proliferation in normal and malignant cells, including adipocytes, monocytes, and tumor cells in colon, prostate, and breast cancers. The current studies were undertaken to assess both the expression and functional activity of PPARgamma in cultured normal human mammary epithelial cells (HMECs) and tissue samples. Analyses by northern hybridization, immunoblotting, and immunohistochemistry demonstrate PPARgamma gene expression in HMECs and breast tissue specimens. DNA binding and transactivation assays indicate the presence of functionally active PPARgamma in HMECs. Treatment with PPARgamma selective ligands, 15-deoxy-delta-(12,14) prostaglandin J2 (15dPGJ2) and ciglitazone, inhibits the growth of HMECs in a dose-dependent manner. This growth inhibition is associated with alterations in cell cycle progression and the induction of apoptosis. PMID- 12725419 TI - Expression of sex steroid receptors and their co-factors in normal and malignant breast tissue: AIB1 is a carcinoma-specific co-activator. AB - The differential expression pattern of estrogen receptor alpha (ER-alpha), estrogen receptor beta (ER-beta) and their co-activator/co-repressor proteins is thought to modulate estrogenic action and to be present already during the early stages of tumorigenesis. It has therefore been postulated that certain co activator and co-repressor proteins contribute to the development of breast cancer. There are some reports providing information on gene amplification and mRNA over-expression of certain co-factors in breast cancer, but to date there is only limited knowledge about their respective protein expressions. The aim of this study was to examine the expression of four steroid receptor co-activators (steroid receptor co-activator 1 (SRC-1), transcription intermediary factor 2 (TIF 2), protein 300 kDa/CREB binding protein (p300/CBP), amplified in breast cancer 1 (AIB1)), and of the co-repressor nuclear receptor co-repressor (NCoR), in malignant breast tissues and in matching normal breast biopsies of the same individuals. Protein expression was analyzed by immunohistochemistry and was compared to prognostic parameters such as lymph node involvement, tumor grading and receptor status. All members of the co-regulatory protein family were detected in both, benign and matching malignant tissue samples, except for AIB1, which was found to be expressed exclusively in malignant epithelium. AIB1 was preferentially present in carcinomas with high tumor grade (r = 0.48, p = 0.014), and was co-expressed with p300/CBP (r = 0.54, p = 0.006). TIF 2 correlated significantly to nodal status (r = 0.46, p = 0.025). Furthermore, protein levels of ER-beta p300/CBP and AIB1 were higher in invasive ductal carcinomas than in normal mammary tissue. The tumoral ER-alpha protein expression was significantly correlated with that of PgR (r = 0.61, p = 0.001) and NCoR (r = 0.4, p = 0.043), whereas ER-beta expression was associated with SRC-1 (r = 0.68, p < or = .001), TIF 2 (r = 0.64, p = 0.001) and NCoR (r = 0.48, p = 0.014) protein levels in malignant specimens. In our hands, 20% of ER-beta positive tumors did not express ER-alpha protein, thereby suggesting that a substantial fraction of ER-beta positive tumors is falsely considered to be 'estrogen receptor negative' if only ER-alpha specific antibodies are employed in the histological assessment of the ER status. PMID- 12725420 TI - Estradiol enhances osteolytic lesions in mice inoculated with human estrogen receptor-negative MDA-231 breast cancer cells in vivo. AB - The effect of 17-beta-estradiol (E2) on the induction of osteolytic lesions by estrogen receptor (ER)-negative breast cancer cells was investigated in 4-week old female nude mice. Exposure to exogenous E2 was found to increase osteolytic areas on radiographs up to 5.3 times in mice inoculated intracardially with MDA 231 human breast cancer cells. The MDA-231 cells were ER-negative, both before inoculation, and after isolation from osteolytic lesions, and the corresponding cell cultures were insensitive to E2. The induction of skeletal lesions by E2 in this mouse model was mainly effectuated at the early development of bone metastases, since exposure to E2 for 8 days around MDA-231 inoculation increased osteolysis to the same level, as did E2 given throughout the entire 31-day experimental period, and because E2-exposure for just the final 14 days had no effect. Independently of exposure to E2, histology revealed cancer cells in hind limp long bones of approximately 80% of the mice, and tumors were absent in non skeletal organs. In vitro studies showed that the number and activity of osteoclasts generated from mouse bone marrow cells were increased 5-6 times when co-cultured with MDA-231 cells. Addition of 0.1-10 nM E2 further dose-dependently increased the osteoclastogenesis and associated bone resorption in these co cultures. In conclusion, E2 was found to increase the morbidity in mice inoculated with ER-negative MDA-231 cells, and to stimulate osteoclast formation and bone resorption in co-cultures of bone marrow cells and MDA-231, suggesting that the progression of osteolytic metastases by ER-negative breast cancer cells can be induced by E2 due to stimulation of osteoclastogenesis. PMID- 12725421 TI - Sulfotransferase 1A1 (SULT1A1) polymorphism, PAH-DNA adduct levels in breast tissue and breast cancer risk in a case-control study. AB - Gene-environment interactions are hypothesized to be major contributors to susceptibility to environmental carcinogens and interindividual variability in cancer risk. We present findings on associations between genetic susceptibility due to inherited polymorphisms of the Phase II detoxification enzyme sulfotransferase 1A1 (SULT1A1), breast cancer risk, and polycyclic aromatic hydrocarbon (PAH)-DNA adducts. A hospital based case-control study was conducted at the New York-Presbyterian Medical Center (NYPMC). The study utilized two control groups: one comprised of women with benign breast disease (BBD) and the other comprised of women visiting NYPMC for routine gynecologic checkups (healthy controls). Blood samples were collected from cases and controls; and breast tissue from pathology blocks was collected from cases (tumor and non-tumor tissue) and BBD controls (benign tissue). PAH-DNA adduct levels were measured by immunohistochemistry in breast tissue samples, and the SULT1A1 (Arg/His) polymorphism at codon 213 was determined by PCR RFLP analyses using DNA from white blood cells. Increasing number of His alleles was modestly associated with breast cancer case-control status, when cases were compared to healthy controls (p for trend = 0.08), when cases were compared to BBD controls (p for trend = 0.08) and when cases were compared to both control groups combined (p for trend = 0.07). Contrary to our hypothesis PAH-DNA adduct levels in breast tissue were not associated with SULT1A1 genotype. Our findings are consistent with a prior report that the Arg/His polymorphism in SULT1A1 is associated with breast cancer risk. PMID- 12725422 TI - Exposures in childhood, adolescence and early adulthood and breast cancer risk: a systematic review of the literature. AB - A growing body of work indicates that exposures over the life course have important roles to play in the aetiology of breast cancer. This review synthesises the literature that has been published in the area of early life events and female breast cancer risk. The review finds some evidence, primarily from cohort studies on the relationship between birthweight and breast cancer, to suggest that in utero events are related to breast cancer risk in adulthood. Strong evidence to support a positive association between height and breast cancer exists. Postulated mechanisms for this relationship include the role of early diet in subsequent disease risk, and the influence of endogenous growth factors mediating the relationship. There is some evidence to suggest that leg length is the component of height which is generating the observed associations between height and breast cancer. There is no consistent pattern of association between relative weight in childhood or adolescence and risk of breast cancer. The evidence to suggest an association between physical activity in early life and breast cancer risk is convincing from case-control studies, but is not fully substantiated by the results of three cohort studies. There are inconsistent results regarding the association between smoking at a young age and breast cancer risk. There is little evidence for an association between passive smoking in early life and breast cancer risk. No clear association between early drinking and breast cancer risk exists. These results are discussed in relation to possible underlying mechanisms and health promotion strategies which could reduce breast cancer risk. PMID- 12725423 TI - Isotopic assessment of sources and processes affecting sulfate and nitrate in surface water and groundwater of Luxembourg. AB - Surface water and deep and shallow groundwater samples were taken from selected parts of the Grand-Duchy of Luxembourg to determine the isotopic composition of nitrate and sulfate, in order to identify sources and/or processes affecting these solutes. Deep groundwater had sulfate concentrations between 20 and 40 mg/L, delta34S(sulfate) values between -3.0 and -20.0 per thousand, and delta18O(sulfate) values between +1.5 and +5.0 per thousand; nitrate was characterized by concentrations varying between < 0.5 and 10 mg/L, delta15N(nitrate) values of approximately -0.5 per thousand, and delta18O(nitrate) values approximately +3.0 per thousand. In the shallow groundwater, sulfate concentrations ranged from 25 to 30 mg/L, delta34S(sulfate) values from -20.0 to +4.5 per thousand, and delta18O(sulfate) values from approximately +0.5 to +4.5 per thousand; nitrate concentrations varied between approximately 10 and 75 mg/L, delta15N(nitrate) values between +2.5 and +10.0 per thousand, and delta18O(nitrate) values between +1.0 and +3.0 per thousand. In surface water, sulfate concentrations ranged from 10 to 210 mg/L, delta34S(sulfate) values varied between -9.3 and +10.9 per thousand, and delta18O(sulfate) values between +3.0 and +10.7 per thousand were observed. Nitrate concentrations ranged from 10 to 40 mg/L, delta15N(nitrate) values from +6.5 to +12.0 per thousand, and delta18O(nitrate) values from -0.4 to +4.0 per thousand. Based on these data, three sulfate sources were identified controlling the riverine sulfate load. These are soil sulfate, dissolution of evaporites, and oxidation of reduced S minerals in the bedrock. Both groundwater types were predominantly influenced by sulfate from the two latter lithogenic S sources. The deep groundwater and a couple shallow groundwater samples had nitrate derived mainly from soil nitrification. All other sampling sites were influenced by nitrate originating from sewage and/or manure. A decrease in nitrate concentration observed along one of the rivers was attributed to denitrification. It appears that sulfate within Luxembourg's aquatic ecosystem is mainly of lithogenic origin, whereas nitrate is often derived from anthropogenic activities. PMID- 12725424 TI - Gas forming processes within lignite mining dumps. AB - Gas analyses of the soil atmosphere of lignite mining dumps yielded increased contents of carbon dioxide. To get information about the potential sources and the carbon dioxide releasing capacity of the dumps, samples of dump material were investigated for their contents and isotopic compositions of organic and inorganic carbon as well as the carbon dioxide in the soil atmosphere. The contents of organic and inorganic carbon were found to vary depending on type of dump material. The isotopic composition of the organic carbon ranges between 24.5 and -26.5 per thousand, which is typical for humous materials. The carbonates are found to be of marine origin (delta13C: +0.5 to -1.1 per thousand). By means of the isotope investigations it could be shown that the carbon dioxide in the lignite mining dump arises from these two different sources. Mixing ratios can be calculated using the isotope balance equation. Both reaction paths are associated with oxygen consumption and do not result in an increased gas pressure within the dump. PMID- 12725425 TI - A new approach to determining the content and 15N abundance of total dissolved nitrogen in aqueous samples: TOC analyser-QMS coupling. AB - The standard method for determining the 15N abundance of total dissolved nitrogen (TDN) in aqueous samples (e.g., soil leachate, sewage, urine) is currently Kjeldahl digestion followed by steam distillation or diffusion to isolate the ammonium, and then 15N measurement using IRMS. However, this technique is both time-consuming and laborious. One way of overcoming these disadvantages could be to couple a TOC analyser to determine the TDN with a sufficient quadrupole MS to determine the 15N abundance. The high TOC analyser (Elementar Analysensysteme Hanau, Germany), which catalytically oxidises the sample's total nitrogen with a high, constant yield to nitrogen monoxide (NO), appeared particularly suitable. The quadrupole-MS ESD 100 (InProcess Instruments Bremen, Germany) proved to be a suitable mass spectrometer for the 15N determination of NO. This combination of instruments was found to provide a workable method in numerous measurements of standard and actual samples. The detection limit concerning the N amount required per analysis is 2 microg, corresponding to an N concentration of 0.7 mg/l in a maximum sample volume of 3ml. Depending on the N concentration, 15N abundances starting from 0.5 at.% can be measured with the required precision of better than 3% (simple standard deviation). For example, measuring the abundance of 0.5 at.% requires about 50 microg N, whereas for 1 at.% or more only about 5 microg N is needed per analysis. PMID- 12725426 TI - Feeding and growth of apple snail Pomacea lineata in the Pantanal wetland, Brazil -a stable isotope approach. AB - Apple snails Pomacea lineata (SPIX 1827) are widespread in the tropical regions of Brazil as well as in the Pantanal wetland of Mato Grosso in the western part of the country. They have a key position in the Pantanal food web and serve as food for many animals e.g. fishes, birds, and caimans. However, little is known about their feeding preferences and growth rates. Stable isotopes have been used successfully on numerous studies as food source indicator. Therefore, the delta15N and delta13C values of snails from 0.45 to 3.03 cm in length, which were collected in the rainy season from March through May, were analyzed. Snails signatures revealed ambiguous evidence for food preferences. Delta15N and delta13C values ranged between -2.8 and 12.4 per thousand and between -24.2 and 16.4, per thousand respectively. This range of values mirrors the highly variable isotope values of possible food sources comprising C3 and C4 macrophytes. To test whether all common food sources were similarly assimilated, feeding experiments with different diets were conducted. Snail eggs were reared in tanks and offered different but single plants. Snails fed different diets and delta13C values of the food were reflected in the animal tissue. Growth varied considerably in experiments with different diets indicating the preference for certain food sources. Also, the fractionation of nitrogen isotopes between food and animal varied from 0.1 to 17.0 per thousand. The results are explained by different feeding habits, and it is supposed that animals fed either on the plant itself or on bacteria mats growing in the tanks. In an additional experiment juvenile snails were offered one single food with a distinctive C4 grass signature. These snails did not grow detectably, but nevertheless isotope signatures approached to values of the diet. PMID- 12725427 TI - Using stable isotope analyses to identify allochthonous inputs to Lake Naivasha mediated via the hippopotamus gut. AB - The hippopotamus grazes nocturnally on land and resides in water during the day. Much of the ingested material must therefore be defecated directly into the aquatic system and can thus be considered an allochthonous resource available to aquatic consumers. The utility of stable isotope analyses of carbon and nitrogen to distinguish hippo faecal matter from other potential basal resources was tested at Lake Naivasha, Kenya. Hippopotami proved faithful to a short grass diet although supplementary grazing of aquatic macrophytes was observed. The typical isotopic ratios of C4 grasses ingested were not altered substantially by gut processes, and were clearly distinct from algal and aquatic macrophyte isotopic ratios. However, marginal plants such as Cyperus papyrus exhibit C4 ratios, and so the technique is suitable only for use in localities where 'contamination' from such sources is negligible. PMID- 12725428 TI - Regime shift signatures from stable oxygen isotopic records of otoliths of Atlantic cod (Gadus morhua). AB - The sudden collapse of Atlantic cod (Gadus morhua) may relate to ocean climate, or regime shifts as demonstrated in production of Pacific salmon. This paper reports the results of stable oxygen isotope ratio analyses (18O/16O or delta18OA) from 91 otoliths of cod over a period of about 20 years. Seasonal delta18OA variations of individual otoliths started at an initial value of about 0.5 to 0 per thousand VPDB, and then reached a stable level in the range of +2.5 to +3.5 per thousand VPDB after 4-5 years. The initial low values correspond to the natal sources of mature cod, while the higher delta18OA values represent the water conditions before the cod was caught. This pattern of delta18OA variation was observed over the life history of all cod examined. Furthermore, the calculated isotopic temperatures agreed with those obtained from summer bottom trawl survey, indicating that delta18OA of otoliths could be used as a thermometer in determining the ambient seawater temperature where the cod lived. Comparison of long-term delta18OA records and biological and meteorological observations suggested that decadal-scale ecosystem changes did occur in the late 1970s and early 1990s in Atlantic Canada, comparable to regime shifts occurred in the North Pacific. PMID- 12725429 TI - Preparation of bone samples in the Gliwice Radiocarbon Laboratory for AMS radiocarbon dating. AB - In the Gliwice Radiocarbon Laboratory, a system for preparation of samples for AMS dating has been built. At first it was used to produce graphite targets from plant macrofossils and sediments. In this study we extended its capabilities with the preparation of bones. We dealt with 3 methods; the first was the classical Longin method of collagen extraction, the second one included additional treatment of powdered bone in alkali solution, while in the third one carboxyl carbon was separated from amino acids obtained after hydrolysis of protein. The suitability of the methods was tested on 2 bone samples. Most of our samples gave ages > 40 kyr BP, suggesting good performance of the adapted methods, except for one sample prepared with simple Longin method. For routine preparation of bones we chose the Longin method with additional alkali treatment. PMID- 12725430 TI - Sulfur isotopic composition of H2S and SO4(2-) from mineral springs in the Polish Carpathians. AB - A number of springs in Carpathian Mts. contain dissolved H2S and SO4(2-) in concentrations above 10 mg/dm3. In this study we have investigated the sulfur isotope composition (delta34S) of the dissolved sulfur species in the springs from the flysch area in the Carpathian Mts. along the tectonic dislocation. It is believed that some of these springs may carry a major fraction of dissolved sulfur species of extremely deep sulfur (of mantle origin), which is subjected to SO4(2-)-H2S isotope exchange at high temperatures. The original isotopic compositions may be modified by reduction/oxidation at low temperatures and by admixture of sulfur from other sources. In order to distinguish the sulfur of mantle origin we investigated delta34S of dissolved sulfide and sulfate and on the basis of known concentrations we calculated delta34S of total dissolved sulfur. The isotope fractionation between sulfate and sulfide helped to distinguish the sulfur origin. Evaluating the sulfur isotope exchange, we selected 4 springs which likely have only weakly disturbed sulfur of mantle origin. PMID- 12725431 TI - Sulphur and oxygen isotopic composition of sulphates in springs feeding the Wieprz river and other springs of Lublin Upland and Roztocze. AB - Springs on Roztocze and Lublin Upland have been studied. Isotopic data are compared with data of chemical analyses. The results of studies allow us to distinguish five types of groundwaters. The differentiation is based upon different lithology; opokas, gaizes, sandy-silty-clay deposits, sands with shell sandstones, marly opokas, marly limestones and 'soft limestones of chalk type. A correlation can be observed between delta34S and the concentration of Ca or Mg ions also a correlation between HCO3- ion concentration and delta18O in sulphates. Probably these correlations are the result of some simultaneous processes, which occur in groundwater. The seasonal variations of the isotopic composition and sulphate concentration were observed in four springs feeding the upper Wieprz. The variations were simultaneous and often similar in these springs. Probably, these variations are caused by the admixture of sulphates coming from shallow water layers (or leached from soil); however the variations of the groundwater level may also change chemical and isotopic composition in groundwater. PMID- 12725433 TI - Colocalization of chromogranin A and inositol 1,4,5-trisphosphate receptor in ciliated cells of the bovine oviduct. AB - Previous investigations of the expression of chromogranin A (CgA) have been performed primarily in neuroendocrine tissues containing amine and peptide secretory vesicles. More recently it has been shown that CgA, as a high capacity Ca2+ storage protein, interacts with the inositol 1,4,5-trisphosphate receptor/Ca2+ channel (InsP3R) which has been found to be selectively localized in oviductal cells of the mouse. To examine a possible role of this coupling in the Ca2+-dependent ciliary movement, we investigated the topographical and cellular distribution of cells positive for CgA and inositol 1,4,5-trisphosphate receptor type 2 (InsP3R2) in the bovine oviduct at different stages of the oestrous cycle. Using immunohistochemical techniques on paraffin-embedded tissue we have successfully shown that CgA is selectively expressed in ciliated cells of the bovine oviduct. The labelled cells show intense positive staining in the apical surface area in close vicinity to the ciliary apparatus. CgA-positive ciliated cells are most frequently observed at dioestrous while a lower number appears at oestrous. Additionally, secretory and intraepithelial neuroendocrine cells consistently do not stain with the CgA-antiserum. We then investigated whether the reported expression of the InsP3R in oviductal cells of the mouse corresponds to the expression of the InsP3R in bovine oviductal cells. Using a polyclonal antibody to the type 2 InsP3R, we found that the receptor is also selectively expressed in a similar matter to CgA in the apical cytoplasm of ciliated cells. This is the first morphological demonstration of the colocalization of CgA and InsP3R in epithelial ciliated cells of the bovine oviduct. Our results suggest that CgA and InsP3R could be involved in controlling the ciliary activity of oviductal epithelial cells. PMID- 12725432 TI - Variations in the cellular proliferation of prolactin cells from late pregnancy to lactation in rats. AB - Lactation is a physiological process associated with hyperactivity of hypophyseal prolactin-producing cells. It is known that the percentage of these cells is increased during lactation, although there are discrepancies in the reports regarding the mechanisms responsible for increasing the number of prolactin cells. In order to analyse whether this increase is a result of previous proliferation, variations in the proliferation rate of prolactin-positive cells were determined from late pregnancy to lactation in adult female rats by means of observation of the immunohistochemical expression of PCNA as a marker of cellular proliferation. During late pregnancy, a very significant increase in the percentage of proliferating prolactin cells was observed in comparison to non pregnant females in the proestrus phase (p < 0.01). Although the percentage of prolactin-positive cells after one week of lactation was higher than in non lactating or in pregnant females (p < 0.01), the proliferation rate was lower than in the other groups studied. In sum, our results suggest that late pregnancy constitutes a preliminary proliferative phase preparatory to the ensuing lactation phase and that endocrine changes in late pregnancy involve the cellular proliferation of hypophyseal prolactin cells in order to prepare the gland for later demands and to prevent proliferative changes from occurring during lactation. PMID- 12725434 TI - Variations in lectin-binding on the zona pellucida during oocyte growth in some wild ungulates. AB - The aim of the present study was to examine the glycoconjugate modifications occurring in the zona pellucida during oocyte growth in fallow, red and roe deer using a battery of lectins combined with sialidase digestion and chemical treatments. This histochemical approach allowed us to sequence the oligosaccharidic side chains of the zona pellucida glycoproteins in these wild ungulates. The most effective lectins in the zona pellucida of these species were SBA, PNA, RCA-I GSA-IB4, and WGA, indicating the presence of beta-D-N Acetylgalactosamine, beta-D-Galactose, alpha-D-Galactose and N-Acetylglucosamine residues. Additionally, sialic acid moieties were demonstrated. We also observed differences in the glycosidic residue content and in their spatial distribution, depending on the species and stage of follicle development. PMID- 12725435 TI - Effects of prenatal procarbazine administration on intrauterine development in rats. AB - The effects of prenatal procarbazine (PCZ) administration on the intrauterine development of rat fetuses were investigated. Gravid rats were treated on day 14 of gestation (GD14) with 25 mg or 50 mg/kg body weight PCZ via stomach tube. Controls received normal saline in the same dosis and manner. On GD20, all fetuses were collected by caesarian section. Live and dead fetuses as well as resorptions were counted. In the live fetuses, the following investigations were conducted: measurement of body weight, occipito-coccygeal-lenght (OCL), tail length (TL), placental weight and diameter, external macroscopic and binocular microscopic examination, and sectional analysis of the animals using the razorblade sectioning technique. Both PCZ doses caused a significant reduction in the number of live fetuses and a significant increase in resorptions. Mean body weight in PCZ groups was antidromic affected. OCL and TL were significantly depressed. Placental weight and diameter as well as number of dead fetuses were comparable to those of controls. External and sectional investigations revealed no PCZ-related deviations. In the light of our findings we conclude that PCZ in the doses used in this experimental study significantly affects the intrauterine development in rats in terms of fetal toxicity but displays no teratological properties. PMID- 12725436 TI - Anomalies in the developing neural and visceral head skeleton of the Australian lungfish, Neoceratodus forsteri. AB - Several anomalies occur in the developing neural and visceral head skeleton of young specimens of Neoceratodus forsteri that have been reared under laboratory conditions. These include anomalies of the basicranium and its derivatives, aberrations of the anterior mandible and hyoid apparatus, and abnormalities in the articulation of the jaws and the elements that produce them. Apart from the occasional absence of the basihyal, and failure of the quadrate processes to form, the anomalies are not deficiencies. Most involve malformations of parts of the neurocranium and visceral skeleton, inappropriate articulations or fusions between elements, disunity in structures that are normally fused and the appearance of supernumerary elements. The incidence of chondral anomalies, generally higher than aberrations that occur in the dermal skeleton in juvenile lungfish, ranges from 1-10% in laboratory reared individuals that have not been subjected to experimental interference. The anomalies differ from those found in many amphibian populations, in the field and in the laboratory, because they involve the cranium, and not the limbs, and the lungfish have not been exposed to the factors that cause anomalies in the amphibians. It is unlikely that the existence of those anomalies, if it is reflected in the wild population, places a selective pressure on the lungfish, because, in a normal season, less than 1% of the total number of eggs produced survive to be recruited into the adult population. PMID- 12725437 TI - Electron microscopic study of intrinsic cardiac ganglia in the adult human. AB - The aim of the present study was to describe in detail the ultrastructure of intrinsic cardiac ganglionic cells in the healthy human as these cells appear to be directly involved in the development of tachycardia, atrioventricular block, ventricular fibrillation, and sudden cardiac death. Tissues examined in this study were obtained from hearts of 10 adult humans of either sex aged 22-80 years at autopsy performed no more than 8 h after death. The examined human intrinsic cardiac nerve cells were in most respects typical autonomic neurons surrounded by a sheath of satellite cells that was either uni- or multilayered. In addition to regular unmyelinated axons, prominent large axon terminals containing lamellated dense bodies, mitochondria and vesicles in the cytoplasm were observed in the ganglion neuropil. Synaptic profiles were more common in the ganglion neuropil than on neuronal somata. According to axon terminal contents, synaptic profiles were of three types. The most common Type 1 synaptic profiles contained a predominance of small clear, with a few larger dense-cored vesicles and mitochondria. Type 2 synaptic profiles, in addition to the same components as in Type 1, had glycogen-like particles. Type 3 vesicle-containing profiles clearly differed from both the previous ones as they were the largest in diameter and included plentifiul large clear pleomorphic or dense-cored vesicles together with small clear and larger dense-cored vesicles, mitochondria, dense and multivesicular bodies. Independently of age of the human, the most frequent neuronal abnormality was an abundant accumulation of inclusions inside of somata and dendrites that, in profile, appeared like circular membranous or fine granular bodies variable in electron density. In addition to inclusions, some neuronal somata and dendrites had strongly swollen mitochondria filled up with granular material in spite of their close association with normal looking ganglionic neurons. Structures resembling an axon growth cone in profile were revealed inside of cardiac ganglia derived from an 80 year old man. In conclusion, the present results provide baseline information on the normal ultrastructure of intracardiac ganglia in healthy humans which may be useful for assessing and interpreting the degree of damage of ganglionic cells both in autonomic and sensory neuropathies of the human heart. PMID- 12725438 TI - A case of bilateral ectopic superior cervical ganglia in man. AB - Bilateral ganglionic masses, likely representing fused superior and middle cervical sympathetic ganglia, were found in the mid-neck region of a cadaver during routine dissection. The displacement of the superior cervical ganglion from its normal location is a striking anomaly that does not appear to have been reported earlier. This observation may be clinically relevant for avoiding misdiagnosis of such masses as Schwannomas or other tumors. In addition, in cases where the superior cervical ganglion is absent from its usual location, it should be sought in the mid-neck region. PMID- 12725439 TI - Distribution of vasoactive intestinal polypeptide in the rat heart: effect of guanethidine and capsaicin. AB - Vasoactive intestinal polypeptide (VIP) is believed to coexist with acetylcholine in postganglionic parasympathetic neurones. However, the presence of VIP in extrinsic nerves and/or other types of intrinsic cardiac neurones has not been excluded. The aim of our study was to examine the distribution and origin of VIP ergic innervation in the rat heart atria using immunocytochemistry and radioimmunoassay (RIA) combined with two types of denervation: sympathectomy, which was produced by guanethidine treatment and sensory denervation achieved by capsaicin administration. In whole-mount preparations of the intact atria, VIP immunoreactive (IR) nerve fibres and ganglionic cells were found, the latter being much more numerous in the left atria (LA) than in the right ones. Some of VIP-IR nerve fibres forming bundles appeared to be extrinsic in origin. VIP-IR concentrations determined by RIA in the intact rats were significantly higher in the LA than in the right ones (p < 0.01). However, no changes in VIP-IR levels were found in either atrium after both guanethidine and capsaicin treatment protocols, thus indicating that VIP-immunoreactivity is not associated with either sympathetic or sensory innervation. In conclusion, the ganglionated plexus of the rat atria may comprise at least 3 different neuronal populations expressing VIP-positivity: 1. extrinsic preganglionic parasympathetic fibres, 2. intrinsic postganglionic parasympathetic neurones and 3. intrinsic local circuit neurones that do not express a cholinergic phenotype. PMID- 12725441 TI - Computed tomography and morphometry of the carpal canal in the dog. AB - This study has been carried out to determine the normal size and shape of the carpal canal in the dog and to describe the salient anatomic structures with the aid of computed tomography. Computed tomographic examination of the carpal region was performed in the left and right forelimbs of 5 male and 5 female healthy German shepherd dogs, respectively. Areas, depths and widths of the carpal canal and identification of the anatomical structures were determined at two levels of the carpal region. The frozen forelimbs of two cadavers were sectioned with a saw to correlate the CT images with these sections. All scans were studied using both bony and soft tissue windows. The measurements of the right and left carpal canals did not differ significantly in male or female dogs, respectively although they were larger in the male than female dogs. In addition to this, the measurements of the carpal canal were larger in the distal than the proximal slice. The lateral tip of the deep digital flexor tendon was located more cranially in the distal part than in the proximal part of the carpal canal. Thus, the distance between the bones and the deep digital flexor tendon decreased and the distance between the deep and superficial digital flexor tendons increased. This localization is important in regard to neuropathy because the median nerve was located between the deep and superficial flexor tendons and this region was wider at the distal than the proximal part of the carpal canal. As a result of this study, it was found that the carpal canal of the dog much resembled that of humans and this similarity can easily generate a predisposition to focal neuropathy namely, carpal tunnel syndrome in dogs. PMID- 12725440 TI - Antigenic profile of human thymus in concurrence with "Clusters of Thymic Epithelial Staining" classification. AB - We examined the expression of various CD coded or not yet defined antigens in human thymus samples using indirect immunoperoxidase and immunoflourescent techniques. Data obtained are presented in concurrence with Clusters of Thymic Epithelial Staining (CTES) classification for various monoclonal antibodies recognizing CD antigens (CD1, CD1a, CD6, CD9, CD14, CD16, CD29, CD30, CD32, CD44, CD45RB, CD47, CD48, CD49a, CD49b, CD49c, CD49d, CD49e, CD49f, CD51, CD53, CD54, CD56, CD57, CD63, CD85, CD95, CD98, CD102, CD103, CD106, CD109, CD146, CD147, CD148, CD151, CD152, CD158a, CD158b, CD164, CD165, CD166) and for monoclonal antibodies 1B10, 5G7, A4, BD46, BLTZ, HP1C5, IND.64, M72, WU947 whose specifities are not yet defined. Some of the mAbs such as CD49f, IND.64 and BD46 are detected as good markers for specific cell types or compartments. Significance of the presence of these antigens on thymic epithelial cells at certain locations is briefly discussed. PMID- 12725442 TI - High origin of the radial artery accompanied by muscular and neural anomalies. AB - A high origin of the radial artery accompanied by muscular and neural abnormalities (three-headed biceps brachii, absence of the palmaris longus muscle, and communication between median and musculocutaneous nerves) were encountered during the dissection of a 25-year-old male cadaver. Co-existence of these variations has not, to our knowledge, been reported in the available literature. PMID- 12725443 TI - Distribution and effects of a single intramuscular injection of India ink in mice. AB - Our goal has been to study the distribution and effects following a single intramuscular injection of a substance using India ink as a tracer. We injected 30 microl India ink in the gastrocnemius muscle group of C57Bl10 mice. Hematoxylin-Eosin, Trichrome stains and polyclonal anti-laminin, anti-collagen-IV and anti-dystrophin were used. The liquid spreads in all directions mainly following the perimysial and epimysial septae. The collagen bundles act as physical barriers preventing passage of the ink particles. In the area of the injection site, necrosis of the fibres is associated with disruption of the basement membrane. In the zones adjacent and distal to the injection site, the liquid progresses by pushing the muscle fibres apart with preservation of the basement membrane. Research based on intramuscular injection of substances should take the following into consideration: a) anatomy of the muscle group injected, b) routes of distribution of the substance, c) types of lesions produced with relation to the site of injection of the liquid, and d) size of the particles of the injected substance. PMID- 12725444 TI - Morphometry of nasal bases and nostrils in Koreans. AB - To investigate morphological characteristics of Korean noses, nasal bases and nostrils were measured and classified in Korean adults. The height and the width of the nasal base, the height and the width of the columella, the length of the long and short nostril axes, and nasal alar angle were measured. The shapes of the nostrils were classified into 7 types by the angle between the right and left long axes of the nostrils. Each measurement was compared with other studies. PMID- 12725445 TI - Incorrect medical terms in terminologia anatomica. AB - We suggest changing the incorrect term of "sinus venosus sclerae" to the exact and logical term of "sinus aquosus sclerae" since this sinus does not contain venous blood, but humor aquosus. We further suggest the name changing "ductus nasolacrimalis" to "ductus lacrimonasalis" signifying thus the exact direction of the flow of the tears. PMID- 12725446 TI - ACS guideline for the early detection of cervical cancer. PMID- 12725447 TI - Anticoagulant use in patients with risk of falling. PMID- 12725448 TI - The importance of taking an occupational history. PMID- 12725449 TI - Is oral sucrose an effective analgesic in neonates? PMID- 12725450 TI - Ambulatory devices for chronic gait disorders in the elderly. AB - Gait disorders in the elderly are common and in most cases cannot be treated medically or surgically. Therefore, treatment often relies on ambulatory devices such as canes, crutches, and walkers. Before selecting a device, the patient should be evaluated to define whether one or both upper extremities are required to achieve balance or bear weight. Patients requiring only one upper extremity can use a cane, while patients requiring both upper extremities are best served by forearm crutches or walkers. The patient's need to bear weight through the device will help the physician choose a specific device. When measuring the device, anatomic landmarks and the angle of the elbow must be taken into consideration. Because time often is limited during a routine office visit, a physical therapist often can provide further training for patients learning to use such a device. PMID- 12725451 TI - Diagnosis and treatment of sick sinus syndrome. AB - Sick sinus syndrome comprises a variety of conditions involving sinus node dysfunction and commonly affects elderly persons. While the syndrome can have many causes, it usually is idiopathic. Patients may experience syncope, pre syncope, palpitations, or dizziness; however, they often are asymptomatic or have subtle or nonspecific symptoms. Sick sinus syndrome has multiple manifestations on electrocardiogram, including sinus bradycardia, sinus arrest, sinoatrial block, and alternating patterns of bradycardia and tachycardia (bradycardia tachycardia syndrome). Diagnosis of sick sinus syndrome can be difficult because of its nonspecific symptoms and elusive findings on electrocardiogram or Holter monitor. The mainstay of treatment is atrial or dual-chamber pacemaker placement, which generally provides effective relief of symptoms and lowers the incidence of atrial fibrillation, thromboembolic events, heart failure, and mortality, compared with ventricular pacemakers. PMID- 12725452 TI - Information from your family doctor. What you should know about sick sinus syndrome. PMID- 12725453 TI - Premenstrual syndrome. AB - Premenstrual syndrome, a common cyclic disorder of young and middle-aged women, is characterized by emotional and physical symptoms that consistently occur during the luteal phase of the menstrual cycle. Women with more severe affective symptoms are classified as having premenstrual dysphoric disorder. Although the etiology of these disorders remains uncertain, research suggests that altered regulation of neurohormones and neurotransmitters is involved. Premenstrual syndrome and premenstrual dysphoric disorder are diagnoses of exclusion; therefore, alternative explanations for symptoms must be considered before either diagnosis is made. The disorders can manifest with a wide variety of symptoms, including depression, mood lability, abdominal pain, breast tenderness, headache, and fatigue. Women with mild symptoms should be instructed about lifestyle changes, including healthy diet, sodium and caffeine restriction, exercise, and stress reduction. Supportive strategies, such as use of a symptom diary, may be helpful in diagnosing and managing the disorders. In women with moderate symptoms, treatment includes both medication and lifestyle modifications. Dietary supplements, such as calcium and evening primrose oil, may offer modest benefit. Selective serotonin reuptake inhibitors such as fluoxetine and sertraline are the most effective pharmacologic agents. Prostaglandin inhibitors and diuretics may provide some relief of symptoms. Only weak evidence supports the effectiveness of gonadotropin-releasing hormone agonists, androgenic agents, estrogen, progesterone, or other psychotropics, and side effects limit their use. PMID- 12725454 TI - Valerian. AB - Valerian is a traditional herbal sleep remedy that has been studied with a variety of methodologic designs using multiple dosages and preparations. Research has focused on subjective evaluations of sleep patterns, particularly sleep latency, and study populations have primarily consisted of self-described poor sleepers. Valerian improves subjective experiences of sleep when taken nightly over one- to two-week periods, and it appears to be a safe sedative/hypnotic choice in patients with mild to moderate insomnia. The evidence for single-dose effect is contradictory. Valerian is also used in patients with mild anxiety, but the data supporting this indication are limited. Although the adverse effect profile and tolerability of this herb are excellent, long-term safety studies are lacking. PMID- 12725455 TI - Screening for cervical cancer: recommendations and rationale. PMID- 12725456 TI - Information from your family doctor. Coronary artery disease: how your diet can help. PMID- 12725457 TI - Information from your family doctor. Coronary artery disease: reducing your risk. PMID- 12725458 TI - Information from your family doctor. Cholesterol. PMID- 12725463 TI - CDC releases guidelines for treating adverse reactions to smallpox vaccination. PMID- 12725464 TI - Effect of occupational exposures on male fertility: literature review. AB - The present review was aimed to determine the influence of working conditions, occupational exposures to potential chemical and physical reproductive toxic agents and psychological stress during work on male fertility. Significant associations were reported between impaired semen parameters and the following chemical exposures: metals (lead, mercury), pesticides (dibromochlorophane, 2,4 dichlorophenoxyacetic acid), ethylene glycol ethers and estrogens. The following physical exposures were shown to deteriorate sperm parameters: radiation (both ionized and microwaves) and heat. Psychological distress has another important contribution to infertility. Several studies indicated that stress has a negative impact on sperm parameters. Occupational parameters should be an important part of history taking among patients attending infertility clinics. PMID- 12725465 TI - Elements in the hair of non-mining workers of a lignite open mine in Neyveli. AB - Trace elements are analyzed in the human scalp hair to assess the extent of body burden of pollution. The levels of seven elements (Cd, Cr, Cu, Mn, Ni, Pb and Zn) were determined in the hair of fishermen from Pondicherry, students and businessmen from Madras and non-mining workers from Neyveli lignite open mine. When compared between them, significantly high concentrations of Cd in the non mining workers from Neyveli and Pb in both the students and businessmen from Madras were observed, thereby indicating environmental source of Cd and Pb pollution. The low Zn level was observed in the fishermen indicating their low nutritional source. In addition to the different residential areas, age, diet, smoking habit and family income of subjects are other factors influencing the concentrations of elements in the hair. PMID- 12725466 TI - A 40-year follow-up of whetstone cutters on silicosis. AB - The present study, consisted of two separate surveys, was initiated to clarify the development of clinical pictures of silicosis after termination of dust exposure. The first survey was a 40-year follow-up of radiographic pictures of the chest among 200 male whetstone cutters (Group I workers). The second survey was conducted in 75 male recipients (Group II workers) of disability compensation for silicosis due to whetstone dust exposure. The study on Group I workers made it clear that the proportion of those free of radiographic findings in the chest pictures decreased during a 40-year follow-up period from 84% in the 1st health examination in 1952-6 to 36% in the 3rd examination in 1995. The rate of progression of the disease from Category 1 to 3 (after ILO-guided classification) to higher categories in a 15-year period was as high as >50%. Longer service duration and higher category of chest radiography at the previous health examination were the influential factors in determining the rapid progression of silicosis. The latter observation was confirmed also through a similar analysis on Group II workers. Whetstone preparations contained SiO2 by about 50%. No industrial hygiene data were available for both groups of stone cutters, but the exposure of Group I workers was estimated to be about 1 mg/m3, or well in excess of the current occupational exposure limit. PMID- 12725467 TI - A proportionate cancer morbidity ratio study of workers exposed to chlorinated organic solvents in Taiwan. AB - We initiated an investigation to examine the possible association between the cancer risk and the chlorinated organic solvents exposure in an electronic factory. To obtain information on the incidence of the various types of cancer among the exposed and comparison groups, the cohort populations were merged with the National Mortality Database, the National Cancer Registry Database, and the National Insurance Hospitalization Database from the Department of Health (DOH), as well as the Labor Insurance Hospitalization Database from the Bureau of Labor Insurance (BLI). The proportionate cancer morbidity ratio (PCMR) was used to estimate the cancer risk of the exposed workers in comparison with either textile workers or electronics workers. After adjustment for age, only the PCMR for breast cancer in the exposed female employees was significantly elevated when compared with the two comparison groups. The increased risk of breast cancer was mainly found in the category of 1989-1997 for year of diagnosis when stratified by calendar year. However, there was no dose-response relationship between female breast cancer risk and duration of employment. Although some PCMRs for the cancers were also increased in the exposed group, female breast cancer was consistently increased when compared with both textile and electronics comparison groups using different exclusion criteria. The results obtained in the present study suggest a possible association between exposure to chlorinated organic solvents and female breast cancer. Since this association has never been reported in the previous studies, further study is needed to clarify the association. PMID- 12725468 TI - Effect of ultraviolet irradiation on the protein synthesis of human skin cells: a study with a monochromatic ultraviolet irradiation apparatus. AB - Human beings always suffer from serious risk of exposure to ultraviolet (UV) rays in general as well as in work environments but the health effects of UV are not yet fully understood. To enable fine analysis of the biological effects of UV at any given wavelength, we developed an apparatus that can experimentally irradiate cultured mammalian cells with monochromatic UV rays at any given wavelength. With this device, the effects of UV at 260, 280, 300 and 320 nm on the total protein synthesis of human skin-derived NB1RGB cells were examined. An inhibitory effect of UV at 260 and 280 nm was evident, whereas UV at 300 and 320 nm had only a marginal effect. UV at 280 nm also decreased amino acid uptake, which appears, at least partly, to be responsible for reduced protein synthesis. Metallothioneins (MTs), known as proteins defending against heavy metal toxicity, have been reported to be induced after UV irradiation, but UV at 280 nm did not induce MTs in NB1RGB cells. Cadmium pretreatment of cells, that had been assumed to protect cells from UV damage via MT induction, did not increase cell viability after exposure to UV at 280 nm. The present study unambiguously revealed the biological effects of monochromatic UV rays, also showing that the UV-irradiating apparatus can be a powerful tool for studying the health effects of UV. PMID- 12725469 TI - Clinical study of asbestos-related lung cancer. AB - We analyzed the characteristics of 120 patients of primary lung cancer supposed to be induced by exposure to asbestos. Most of 120 patients were male and the age ranged from 47 to 87 years with a median of 70 years. No particular tendency was observed in the histological types of the lung cancer in 120 patients. Forty of the 120 patients were heavy smokers. When the occupational history was analyzed, most of the patients had been exposed to asbestos in former Japanese naval shipyard, commercial shipyards, construction industry and ironworks. The term of asbestos exposure was 2 to 60 years with a median 27 years. Lung cancers appeared after 15 to 69 years with a median 43 years from the initial exposure to asbestos. Lung cancer was accompanied by asbestosis in 35 patients and by pleural plaques in 77. Twenty-two patients had both asbestosis and pleural plaques. The number of asbestos bodies per 5 g wet lung tissue for 72 patients whose lung tissues obtained from autopsy or surgery was more than 150 bodies which meant the number of occupational asbestos exposure. As for the kinds of asbestos fibers of 32 patients, 14 patients exposed to crocidolite, 10 patients to amosite and 8 patients to chrysotile. PMID- 12725470 TI - A prospective observation of onsets of health defects associated with working hours. AB - A prospective study examined whether working 50 hours or less per week was associated with an increase of mental and circulatory disorders, and if so, whether it was significant for managing workers' health. White-collar workers aged 20 to 60 working at an office of a Japanese company were surveyed from August 1997 to March 2001. The onsets of the diseases were confirmed using medical insurance claims, and the overall incidence rates were 6.6 and 15.7 per 10,000 person months for both the disorders, respectively. Though not significant, those who worked 45 hours or more showed higher relative risks for mental disorders compared with those who did not. No other analysis indicated distinct findings. The small number of onsets of the diseases, the attention to personal health for workers who worked shorter hours, and intensive health care system for circulatory diseases in the workplace may partly account for the findings above. However, through reviewing other reports, it was suggested that the perception of work time could disturb the association between these findings, especially for white-collar workers. Also, it might be useful to observe the onset of mental disorders in work of less than 50 hours with the workers' perception of work time. PMID- 12725471 TI - Additive effect of smoking and cotton dust exposure on respiratory symptoms and pulmonary function of cotton textile workers. AB - One hundred and sixty-nine and 175 cotton textile workers (CTWs) were enrolled in the first (1991) and second (1996) surveys to investigate the prevalence of byssinosis. The synergistic effect of smoking on cotton dust exposure was also evaluated. Although the difference in prevalence of abnormal pulmonary function between the first (38.5%) and second study (38.9%) was not statistically significant, smokers had significantly higher frequency than nonsmokers in both surveys. A significant trend existed between the cotton dust levels and the frequency of abnormal lung function. The significant trend was also noted in both smokers and nonsmokers. The frequency of respiratory symptoms and the prevalence of severe byssinosis in the second survey (14.9% and 12.6%, respectively) were significantly lower than that in the first survey (39.7% and 21.9%, respectively). The reduction of symptoms was due to remodeling of this old cotton mill. The prevalences of respiratory symptoms and byssinosis in smokers being significantly higher than in nonsmokers only found in the first survey, but not found in the second survey. These results indicate that smoking potentiates the effect of cotton dust exposure on respiratory symptoms and byssinosis. The second study reveals high prevalence of byssinosis still existed in Taiwanese cotton mill, although the prevalence was declining. Smoking was found to show an additive effect on cotton dust exposure. Anti-smoking campaign, occupational health program to reduce the dust exposure, and periodical medical examination are measures to prevent from byssinosis. PMID- 12725472 TI - A comparison of white-collar jobs in regard to mental health consultation rates in a health care center operated by a Japanese company. AB - The aim of this study was to compare the consultation rates (CR) of workers performing several types of white-collar jobs. We collected data from the database inputted at the first consultation to a health care center from April 1996 to March 1999. We found that the CR of engineer group was 2.3 times higher than that of employees involved in research and development group. We speculate that this is partially due to the portion, which is not small, of computer systems engineer (SE) comprising this group; SE has previously been mentioned as a particularly stressful occupation in Japan. Since the result of this study is preliminary, we need to conduct a further study taking into account that multiple factors affect CR. PMID- 12725473 TI - Occupational respiratory diseases in the Czech Republic. AB - This paper presents the profile of occupational respiratory diseases in the Czech Republic. In a retrospective study the author analyzes structure, causes, occurrence, and trends of occupational diseases. Between 1996 and 2000, a total of 2,127 new cases were recorded, of which 62.0% were pneumoconioses caused by dust containing free silica, 21.0% were occupational asthma or allergic rhinitis and the rest were divided between lung cancer (10.0%), asbestos-related disorders (4.4%) and variety of other respiratory diseases (2.7%). During the period of the investigations, the decreasing trend of occupational respiratory diseases, which began in 1992, has continued. PMID- 12725474 TI - Training in paediatric palliative medicine. PMID- 12725475 TI - The Cochrane Pain, Palliative and Supportive Care Group: scope and challenges. PMID- 12725476 TI - Bereavement research: methodological issues and ethical concerns. AB - Principles of conducting research in the field of bereavement are introduced. The review focuses on issues of design and methodology, and considers ethical concerns, particularly in view of the fact that bereaved people are frequently in acute grief when scientific study is conducted. Accurate assessment is fundamental, and requires methodologically stringent procedures. The merits and disadvantages of different types of investigation are considered, the often neglected need for control groups is described, and quantitative versus qualitative methodological approaches are reviewed. Also considered are biases due to selection and limits on generalizability. Understanding of ethical matters arising in this field of research is essential for the conduct of worthwhile research, and involves methodological sophistication. PMID- 12725477 TI - The experiences of donor families in the hospice. AB - OBJECTIVES: This study explored the experiences, attitudes and feelings of relatives who consented to donation of corneas of a loved one who had died of cancer. DESIGN: Semi-structured interviews were conducted by a single researcher and qualitatively analysed for systematic identification of themes. SETTING AND PARTICIPANTS: Two specialist palliative care units in Southern England participated. The names of donor families were obtained via transplant coordinators, the local eye bank and hospice records. Ten donor families were interviewed, four to 12 months after their relative's death. RESULTS: The majority of interviewees had thought the deceased was ineligible for donation due to cancer or old age. Approaches regarding donation were generally made by nurses and described as sensitive by relatives. Very few family members would otherwise have considered or raised the subject. Almost all were glad they had agreed to donation although few felt it directly affected their bereavement. A range of motivations for consent was encountered but decisions were very much easier if the deceased's wishes were already known. Most favoured an opt-out system or the right of the family to decide if the deceased's wishes were not known. CONCLUSIONS: Corneal donation is possible for patients dying from cancer. This study indicates that it is generally regarded positively by donor family members. Education of the public and health professionals is necessary to highlight both the eligibility criteria and the need for donation. Families should be encouraged to discuss donation to facilitate respecting individuals' wishes. In the absence of a known decision from the deceased, health professionals should be aware of their responsibility to discuss the issue with the family. PMID- 12725478 TI - Intravenous morphine for rapid control of severe cancer pain. AB - This randomized controlled trial compared intravenous route with oral route for initial dose titration of morphine in 62 patients with end-stage cancer and severe pain. Patients in the intravenous group received 1.5 mg intravenous bolus doses of morphine every ten minutes till pain relief was total or until they became drowsy. After that they got oral morphine at a dose equal to the total initial intravenous requirement four-hourly. Patients in the oral group got oral morphine 5 mg doses (if opioid-naive) or 10 mg (if already on weak opioid) four hourly. Patients in both groups had the option to receive rescue doses of their regular oral dose as and when needed, if necessary hourly. Twenty-seven of 31 in the intravenous group had either total or satisfactory pain relief by the end of one hour, whereas only eight of 31 in the oral group had a similar result. After 24 hours and later both groups had similar results. There was no immediate serious side effect in any of the patients. The late side effects were similar in the two groups. In the intravenous group, the ratio of initial intravenous dose requirement to the subsequent regular single oral dose after two days centred around 1:1 (range 1:0.5-1:3.3). This study found the intravenous method to be safe, effective and superior to the traditional method in providing immediate relief to severe cancer pain. PMID- 12725479 TI - Octreotide for palliative treatment of hepatic metastases from non-neuroendocrine primary tumours: evaluation of quality of life using the EORTC QLQ-C30 questionnaire. AB - BACKGROUND AND AIM: While octreotide has been used in palliative treatment of hepatocellular carcinoma and neuroendocrine tumours with good results, little is known about the possible role of this in palliative treatment of hepatic metastases. MATERIAL AND METHODS: We present our experience from the use of octreotide in palliative treatment of symptomatic liver metastases in 16 patients (11 males, five females, age ranged 43-69 years) with proven hepatic metastases from different primary tumours (six with non-small lung cancer, four with colon carcinoma, two with primary pancreatic head carcinoma, two with prostate cancer and two with adenocarcinoma of the stomach). All patients were administered 20 mg long-acting octreotide IM (octreotide LAR) once the first day, octreotide SC 0.5 mg three times daily on days 2-14 and then 20 mg long-acting octreotide IM every month. Quality of life was assessed by using the European Organization for Research and Treatment of Cancer (EORTC) Core Quality of Life Questionnaire (QLQ C30). Tumour response was evaluated by using ultrasonography. RESULTS: One month after baseline, octreotide resulted in significant (P < 0.01, Wilcoxon test) improvement and stabilization of all major related EORTC QLQ-C30 parameters such as global quality of life, pain, fatigue, insomnia, appetite loss as well as physical, emotional, cognitive, social and role functioning. Except for mild hyperglycaemia in six out of 16 patients and mild gastrointestinal complications in one patient, no other severe side effect due to octreotide was reported. Two patients died two months after the initiation of the study due to generalized metastatic disease, while the remaining 14 patients were still alive seven months after the initiation of the study. The hepatic metastases were stabilized and no new lesions were detected by ultrasonography. CONCLUSIONS: Although further studies are warranted, we consider the use of octreotide a good alternative in palliative treatment of symptomatic liver metastases in patients with end-stage malignant disease. PMID- 12725480 TI - An exploration of dignity in palliative care. AB - This paper presents a qualitative study exploring the meaning of 'dignity' to patients, relatives and professionals. It examines the impact of advanced illness and treatment and the issues pertinent to caring for dignity. Depth interviews were conducted with eight patients, six relatives and seven members of the multiprofessional team. A phenomenological approach to data analysis was adopted. Dignity was found to be a complex phenomenon. It is composed of the dimensions 'being human', 'having control', 'relationship and belonging' and 'maintaining the individual self'. These dimensions seemed to be held in equilibrium by each individual. The importance of each may alter in response to threats to dignity such as advancing illness and how one is treated within it, in an attempt to cope and adapt. Caring for dignity is challenging because it involves balancing the multiple needs of both users and providers of palliative care and the different perceptions and dimensions of dignity itself. PMID- 12725481 TI - The development of palliative care in national government policy in England, 1986 2000. AB - Palliative care is an established and expanding speciality, important in many areas of service delivery within health and social services and the voluntary sector. Traditionally, palliative care is viewed as most closely linked to cancer services. National government policy has an inevitable impact on the organization and provision of such services. As part of a wider project, an investigation of the content and development of English government policy relating to palliative care was carried out. The development of policy follows a cycle that has no closure; it takes place within changing contexts, between multiple organizations and across sectors. Data collection involved the collation and close examination of policy documents and other documents that demonstrably influenced policy. This was in order to identify a national view of policy intentions for palliative care. In total, 53 policy documents were examined. The analysis revealed a number of pertinent issues relating to shifts in funding policies and an evolving definition of palliative care. There is an increasing recognition in the policy documents of the need for palliative care to extend beyond the traditional focus on cancer services and terminal illness. In tandem, is an increasing recognition of the need for partnership between sectors, reflected in more recent health and social services legislation. PMID- 12725482 TI - A stakeholder evaluation of the impact of the palliative care clinical nurse specialist upon doctors and nurses, within an acute hospital setting. AB - There has been an expansion in the number of palliative care teams based in the acute hospital setting. Although organization of these teams varies both in structure and approach, the clinical nurse specialist is one of the key members. The last decade has seen an escalation in the UK of clinical nurse specialists, and following the Calman-Hine Report and the more recent NHS Cancer Plan, it is anticipated that the number of clinical nurse specialists in palliative and cancer care is likely to grow. This paper presents the qualitative findings of an evaluation study designed to investigate the impact of the clinical nurse specialist within a palliative care team based in an acute hospital setting. A stakeholder evaluation that encompassed 31 tape-recorded semi-structured interviews with senior nurses, consultants, junior doctors and nurses representing different grades were performed. The data was analysed for emerging themes utilising a case and cross case analysis methodology. The results suggested the presence of the clinical nurse specialists is seen as beneficial to both medical and nursing staff. This paper will focus upon exploring these potential benefits, included providing support, advice and education. PMID- 12725483 TI - Dying in the community: general practitioner treatment of community-based patients analysed by chart audit. PMID- 12725484 TI - A scanning electron microscopic study of phenotypic plasticity and surface structural changes of aortal smooth muscle cells in primary culture. AB - Phenotypic modulation of smooth muscle cells (SMCs) from a contractile to a synthetic state characterised by active proliferation appears to be an early event in the pathogenesis of atherosclerosis. A similar transition occurs when SMCs are established in culture. In this study the phenotypic plasticity and surface structural changes of aortal smooth muscle cells during the transition from the contractile to the synthetic state and during maturation have been structurally assessed by scanning electron microscope (SEM). The experiments were performed on SMCs obtained from aorta of neonatal rats after enzymatic digestion and then cultured on glass coverslips. SEM observations revealed a three dimensional appearance characteristic for different stages of SMCs. Intensively proliferating cells from monolayer region were large, polygonal in shape with lamellipodia and well spread. Long, uniform in diameter, finger-like microvilli were densely arranged on the surface of these cells. In the thickened region of culture, the cells were rather small, generally spindle-shaped, not well spread, with low density of short, bubble-like microvilli on the surface. Numerous plasma membrane structural alterations in apoptotic cells were observed by SEM: loss of cellular adhesion, smoothing, shrinkage and outpouching of membrane segments have been recognised as markers associated with the cell injury and death. It was concluded that scanning microscopy observations would allow a more complete understanding of SMCs and their changes in culture and atherosclerotic disease. PMID- 12725485 TI - Morphology of the temporal canal and postglenoid foramen with reference to the size of the jugular foramen in man and selected species of animals. AB - The jugular foramen and postglenoid foramen are the main venous foramina of the skull of placental mammals. Their mutual relations are closely related to the development of the internal and external jugular vein, depending on the given kind. On the basis of measuring studies, it was decided to investigate the relations of the size of these foramina and to quantitatively determine which of them prevails. The studies were performed with macerated human and animal skulls. Altogether 100 skulls of human adults of both sexes, 100 of macaccas, 67 of bisons, 25 of mongrel dogs, 37 of foxes and 25 of rats were examined. The jugular foramen was the outlet of the sigmoid sinus or its equivalents and the postglenoid foramen--the petrosquamous or temporal canal. The jugular foramen was always singular and characterised by a small variability in the morphology, consisting only in the occurrence of the internal division or its lack. The postglenoid foramen, on the other hand, in the majority of the studied kinds was variable and numerous. The number of foramina ranged from 1 in man to 7-10 in bisons. In the course of measurements, it was found that the area of the postglenoid foramen in relation to the area of the jugular foramen is 0.87% in man, 24% in macaccas, 30.7% in dogs, 34.4% in foxes, 53.9% in rats and 248.5% in bisons. The results obtained indicate that in bisons the system of venous sinuses, connected with the postglenoid foramina, has a decisive significance for the drainage of the cranial cavity. In contrast, the postglenoid foramen in man has a marginal significance. The remaining kinds, whose skulls have been investigated, occupy on that score an intermediate position. PMID- 12725486 TI - Immunocytochemical evidence for growth hormone-releasing hormone in the tanycytes of the median eminence of the rat. AB - The current study was performed to analyse the potential existence and structure of a GHRH-transporting tuberoinfundibular system in the rat median eminence. The immunocytochemical analysis using anti-GHRH revealed an intense immunoreaction in the ependimary cells, tanycytes, at the level of the floor of the infundibular recess forming part of the median eminence. The basal processes of these cells course towards the external layer of the median eminence and reach the growth hormone-releasing hormone (GHRH) fibres of the tuberoinfundibular tract and this reaction was increased after intraventricular treatment with colchicine. Thus, these observations suggest the existence of a second or alternative cerebrospinal fluid-mediated route of GHRH transport to the median eminence and implicate the involvement of tanycytes in the regulation of this novel transport system. PMID- 12725487 TI - Co-existence of apoptotic and necrotic features within one single cell as a result of menadione treatment. AB - In the present study we examined the effects of menadione, a redox cycling agent, on structural changes of human osteosarcoma line 143B cells. It has been previously reported that menadione can cause necrotic or apoptotic cell death in a concentration- depending manner. In our experimental model, cells were treated with 100 microM menadione for 24 hours. Using electron microscopy technique cells carrying three kinds of morphological changes were detected: necrotic cells, apoptotic cells and those demonstrating a co-existence of apoptotic and necrotic features in one single cell. PMID- 12725488 TI - Distribution of nitric oxide synthase and neuropeptide Y neurones during the development of the hippocampal formation in the rat. AB - Nitric oxide (NO) is a short-lived radical, which modulates synaptic plasticity, neuronal oscillations and cerebral blood flow. NOS-containing neurones can be detected anatomically by nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry or by NOS immunohistochemistry. Neuropeptide Y(NPY) is the most abundant peptide in the brain. NPY is connected with several vital functions, such as a feeding behaviour, sexual maturation, regulation of circadian rhythms, body temperature, blood pressure and neuroendocrine secretions. Neuropeptide Y also modulates anxiety-related disorders, limbic epileptic seizures as well as learning and memory processes. The study was performed on 45 Wistar rats of various ages (PO, P4, P7, P10, P14, P21, P30, P60, and P120; P--postnatal day). The free-floating sections were stained with standard immunohistochemistry methods. Thereafter the histological sections were studied using the confocal laser microscope equipped. For 3D reconstruction the image analysis program LaserSharp 2000v. 2.0 (Bio-Rad, UK) was used. We found that in the newborn rat both NOS- and NPY-immunoreactivity was weak. It had been increasing gradually until the 7th day of postnatal life, after that until P14 it was maintained on the similar level, and then the number of immunolabelled cells deceased. The developmental changes concerned cell morphology as well--until the 10th day of life the immunoreactive cells were immature, with round or oval bodies and had only a few fibres. From P14 the cells' morphology became similar to that in adult. PMID- 12725489 TI - Differentiation of the cortical plate in early human foetuses. AB - The aim of the study is to describe the differentiation of the cortical plate in human foetuses aged 9-11 weeks. Histological sections showed that in this early period of development the cortical plate is differentiated into two zones. The external zone is composed of vertical columns of cells, which are perpendicular to the cortical surface. The internal zone is wider and consists of irregularly arranged clusters of cells. PMID- 12725490 TI - Mast cell density, neuronal hypertrophy and nerve growth factor expression in patients with acute appendicitis. AB - In acute appendicitis, although the relationship between the enteric nervous system (ENS) and mast cells (MCs) has been described in a few studies, neither the expression of nerve growth factor (NGF) nor its relation to mast cell density (MCD) and ENS has been delineated yet in this disease. The aim of this study was to immunohistochemically investigate the relationship between MCD, nervous system and NGF expression in the appendices of cases with clinically and histopathologically diagnosed acute appendicitis and of normal controls. Twenty five patients with acute appendicitis and twelve normal controls were included in our study. Mast cell tryptase, PGP 9.5 and anti-NGF immunostained tissue sections were subjected to quantitative image analysis. Our results showed that MCD, the number of Schwann cells, the number and size of ganglia and NGF staining were significantly greater in acute appendicitis than in the control group (p < 0.01). A strong correlation between MCD and NGF staining was detected (r = 0.92) only in cases with acute appendicitis. Similarly MCD was also related to neuronal proliferation and hypertrophy in this group. We failed to detect any relationship between NGF staining and neural components either in the acute appendicitis or control groups. Our findings indicate that mast cells could be one of the important cell populations responsible for nerve proliferation and hypertrophy in acute appendicitis. The relationship between NGF staining and MCD and the lack of correlation between NGF staining and changes in neural components suggest that, in acute appendicitis, NGF might be responsible for the increased number of MCs, but not for neuronal proliferation and hypertrophy. PMID- 12725491 TI - Formation of synapses on the growth cones in human embryonic inferior ganglion of the vagus. AB - Ultrastructural study was performed on inferior ganglia of the vagus in human embryos aged 7and 8 weeks (developmental stages 18 to 23, 44 to 56 days). The growth cones are observed between the bundles of axons of the inferior ganglia of the vagus. Many primitive synapses (protosynapses) between dendritic and axonal growth cones are observed. PMID- 12725492 TI - The neuronal structure of the globus pallidus in the rabbit--Nissl and Golgi studies. AB - The studies were carried out on the telencephalons of 12 adult rabbits. Two types of neurons were distinguished: 1. Large neurons (perikarya 18-40 microm), which have from 2 to 6 thick, long primary dendrites. Their perikarya have a polygonal, triangular and fusiform shape. The large neurons in the centre of GP have radiated dendritic trees, whereas the dendritic field of the cells along the borders of GP has an elongated shape. The dendritic arbour is not homogeneous. The dendrites may be covered with spindle-shaped dendritic swellings, bead-like processes, not numerous spines or they may be smooth as well. The dendritic branches form thin, beaded dendritic processes, that arise from any part of the dendritic tree, as well as "complex terminal endings" which have various types of appendages on their terminal portions. An axon emerges from a thick conical elongation either from the cell body or one of the dendritic trunks. These neurons are the most numerous in the investigated material. 2. Small nerve cells have been infrequent in our material. Their cell bodies are rounded or polygonal. From the perikarya there arise 2-4 thin dendritic trunks, which may have irregular swellings and few spines. The dendrites spread out in all directions, making the dendritic field round or oval in shape. Generally most axons of the small cells have not been impregnated. However, a few of them have a thin axon with a conical elongation, which emerges from the cell body and bifurcates into beaded processes. PMID- 12725493 TI - Computed tomographic assisted study of morphological changes in the sutural areas as resulting from obliteration. AB - The authors have studied the morphological differences of sutural areas in human skulls at distinct stages of obliteration. A computed tomography aided with computer algorithm of tonal correction was applied to visualise structural changes in the sutural areas that are correlated with sutural ossification. The results of our study suggest that obliteration of cranial sutures is accompanied by local fluctuations of bony density in sutural areas, gradual unification of structural density towards ectocranium. When the obliteration proceeds, the sutural gap ceases. The laminas and diploic layers of the two opposing bones become a single structure of increased integrity. PMID- 12725494 TI - Divergent effects of quinolinic aminoxyls on mitochondrial ultrastructure and localisation in osteosarcoma 143 B cells. AB - In the present study we have shown that quinolinic aromatic aminoxyls are very efficient in protecting lipids of endoplasmic reticulum membranes against hydroperoxide-induced oxidation. The efficacy of these aminoxyls as protectors of lipids was much higher than the water-soluble 4-OH-TEMPO. We have also shown that QAL causes distinct changes of the morphology of mitochondria: from filamentous to granular enlarged structure via the folding of the former. QAL induces also perinuclear clustering of mitochondria. C-QAL as well as 4-OH-TEMPO treated cells revealed filamentous and scattered pattern of mitochondria. Antioxidant activity of QAL as well as morphological changes of mitochondrial raise the possibility that this drug can affect cell physiology via changes of mitochondrial function. PMID- 12725495 TI - Arterial supply of the olfactory trigone and the anterior perforated substance in macrosmatic and microsmatic animals. AB - An investigation was conducted into the differences between macrosmatic and microsmatic animals of arterial supply in the area of the olfactory trigone and the anterior perforated substance. A brain of domestic cat was taken as an example of a macrosmatic animal and a brain of baboon as an example of a microsmatic animal. The cerebral blood vessels of the cadavers of 30 cats and 11 baboons were filled with latex and, after fixation in acid alcohol by means of microscopic operation, the cortical and deep branches of the anterior and middle cerebral arteries were dissected. We discovered that there were differences between these two groups of animals in the places of arising of deep branches and in their course. In the cat the deep branches running through the area of the olfactory trigone arose from the beginning part of the middle cerebral artery in numbers 5-10 and entered the brain in the olfactory tubercle. In the baboon the deep branches arose from two sources: the middle cerebral artery and the anterior cerebral artery. The branches of the middle cerebral artery went through the anterior perforated substance, forming two groups of branches: lateral and medial. PMID- 12725496 TI - The neuronal structure of the medial geniculate body in the pig--Nissi and Golgi study. AB - The studies were carried out on the brains of adult pigs. The preparations were made by means of the Golgi technique as well as the Nissl and Kluver-Barrera methods. Four types of neurons were described in the medial geniculate body (MGB) of the pig: 1. Multipolar neurons (perikarya 30-45 microm) with rounded, oval or quadrangular perikarya from which arise 4-7 dendritic trunks. The dendrites divide dichotomically twice, may send out collaterals and give off ramifications. The dendritic branches possess varicosities and knob-like spines. These neurons predominate in MGB. 2. Pear-shaped neurons (20-35 microm) with one or two dendritic trunks arising from one pole of the cell body. These dendrites have a tufted appearance. 3. Triangular neurons (30-45 microm) possess three thick dendrites which first bifurcate near the soma and then divide profusely into daughter branches. 4. Fusiform neurons (30-50 microm) have usually two dendritic trunks which arise from the opposite poles of the cell body and divide dichotomically twice. The fusiform neurons are the least numerous in MGB. Most MGB neurons have on the secondary tertiary dendrites and on their ramifications have delicate varicose or bead-like appendages and spine-like protrusions. In all types of neurons an axon arises either from the soma or from the initial portion of the dendritic trunk. PMID- 12725497 TI - Sciatic nerve variations in some studies on the Polish population and its statistical significance. AB - The purpose of the study was to describe and analyse sciatic nerve variation in a relatively small, yet statistically significant, group. Consequently, a comparison of described variation to that known from former studies was performed. Additionally, we estimated the minimum group necessary for a statistical confirmation of hypothetical differences in sciatic nerve topography between the studies. PMID- 12725498 TI - The arrangement of muscle fibres in the region of the subthebesian fossa in the aspect of atrial flutter. AB - The typical atrial flutter is the most common supraventricular tachycardia with reentrant circuit. This tachyarrythmia is based on macroreentry wave going around established anatomic landmarks. The reentry in the inferior right atrial wall passes through narrow isthmus, which is the goal for ablative therapy. The isthmus area is bordered anteriorly by the tricuspid valve and posteriorly by the inferior vena cava, coronary sinus, and eustachian ridge. Near to this area we can find anatomical structure, which can be very important during arising, perpetuation and curing of atrial flutter. The concavity, so-called subthebesian fossa, is on the way of typical atrial flutters' reentrant circuit. Regarding the facts mentioned above we decided to examine the morphology and the arrangement of the muscle fibres in this fossa. Research was conducted on material consisting of 70 human hearts of both sexes from the age of 34 to 72 years. 50 hearts came from patients whose death was not cardiologic in origin. 20 hearts came from humans in whose common atrial flutter was confirmed. We observed the arrangement of muscle fibres in the area of subthebesian fossa. Besides we measured the size and deepness of the subthebesian fossa in both groups of hearts. We found that regular arrangement of muscle fibres within subthebesian fossa was present in 23 healthy human hearts (46%) and 7 cases (35%) of hearts with atrial flutter. The irregular arrangement of muscle fibres was observed in 27 hearts (54%) of control group and 13 hearts (65%) with dysrrhythmia. The thickness of the right atrial wall within the subthebesian fossa was very thin in 8 normal hearts (16%) and in 5 dysrrhythmic hearts (25%). The sizes of examined structure were variable in both groups of hearts, and are presented in the table. It seems that the subthebesian concavity can be the substrate for reentrant circuit during atrial flutter, and there could be such special arrangement of muscle fibres, which allows for microreentrant circuit to arise in this area. PMID- 12725499 TI - Localisation of focal liver lesions to specific hepatic segments--comparison of multiphase spiral CT and MR imaging. AB - The purpose of this study was an evaluation of the ability of the multiphase spiral CT and MR imaging to localise focal liver lesions referring to specific hepatic segments. The authors studied prospectively 26 focal liver lesions in 26 patients who had undergone spiral CT and MRI before surgery. Multiphase spiral CT included non-contrast scans, hepatic arterial-dominant phase, portal venous- dominant phase and equilibrium phase. MRI was performed in all cases. The following sequences were performed: SE and TSE T1- and T2-weighted images, STIR and dynamic T1-weighted FFE study after i.v. administration of gadolinium (Gd DTPA). The CT and MR scans were prospectively and independently reviewed by three radiologists for visualisation of hepatic and portal veins and segmental localisation of hepatic lesions. The authors used the right and left main portal veins along with transverse fissura, hepatic veins and gallbladder fossa as landmarks for the tumour localisation to specific hepatic segments. The primary segmental locations of the lesions were correctly determined with CT in 22 of 26 focal liver lesions (85%) and with MR imaging in 24 of 26 lesions (92%). The full extent of lesions was correctly described with sCT in 19 of 26 focal lesions and with MR in 21 of 26 tumours. MRI and CT were helpful preoperative tools for determining the segmental location of focal liver lesions and for planning the surgical approach. PMID- 12725500 TI - The neuronal structure of the inferior colliculus in the bank vole (Clethrionomys glareolus)--Golgi and Nissl studies. AB - The inferior colliculus (IC) of the bank vole is made up of 3 nuclei: the external and pericentral nucleus, which are located on the outer border of the IC, and the central nucleus, which is the largest part of IC and shows a laminated structure. On the basis of various morphological criteria 5 types of neurons have been distinguished in the bank vole IC: 1. The rounded cells (perikarya 10-15 microm) with 2-4 primary dendritic trunks. The dendritic tree has a spindle-like shape. The axon emerges from the soma or from the proximal portion of a dendrite. 2. The fusiform neurons (17-20 microm) with 2 primary dendrites arising from both poles of the perikaryon. The dendritic tree has the same shape as the previous type. The axon originates from the proximal dendritic trunk. The rounded and fusiform cells constitute the main neuronal type. 3. The pear-shaped neurons (10-13 microm) with 2 main stems or rarely 1. The axon emerges from the perikaryon or seldom from the dendritic trunk. 4. The multipolar cells (18-23 microm), which have from 4 to 6 primary dendrites radiating in all directions. The dendritic tree has a spherical shape. The axon emerges either from the proximal stem or directly from the soma. 5. The triangular neurons (15 18 microm) with 3 primary dendritic trunks. The axon originates from the perikaryon. The triangular cells are the least numerous. All types of neurons in the bank vole IC bear spines and protrusions. PMID- 12725501 TI - Analysis of operating field area in transpyramidal retrolabyrinthine approach to posterior cranial fossa. AB - Retrolabyrinthine surgical access to the posterior and middle cranial fossa has a long history of use during the procedures aiming at the removal of small neo plasmatic changes located in the area of the internal auditory tube. A precise knowledge of the anatomical alterations of the temporal bone in the aspect of the retrolabyrinthine access to the posterior cranial fossa determines a successful otoneurosurgical endoscopy, which involves a relatively narrow area. Forty-four cadaver temporal bones of both sexes were measured to obtain the dimensions of the surgical area limited by the sigmoid sinus, superior petrosal sinus and posterior labyrinth. The techniques of computer picture analysis were applied in the research. The mean value of the surface area of the figure limited by the sigmoid sinus, superior petrosal sinus and posterior semicircular canal was 175.9 mm2 with no significant differences between sexes and sides. The maximal measured value was 356 mm2, and the minimal was 84.3 mm2. The size of the surgical area is characterised by large deviation range but was always sufficient to insert the endoscopic device and standard otosurgical instruments. PMID- 12725503 TI - Application of a mixture of glycol polyethylenes for the preparation of microcorrosion casts--an observation. AB - Preparation of microcorrosion casts that can be used for observation in SEM is a laborious, time-consuming procedure. The authors paid particular attention to the process of dissection of the microcorrosion casts. This prompted the authors to reconstruct the plastic mass, produced by the firm Gurr (Great Britain) in the 1970s, which was used by them in previous research to immerse the cast in order to minimise the damage. By using easily obtainable polyethylene glycols, characterised by different physical and chemical features, in order to obtain smooth surface of the section, a low-toxic mixture was composed, which protected the microcorrosion casts sufficiently and did not interfere with the physical and chemical properties of the cast. PMID- 12725502 TI - Intravascular ultrasound assessment of blood vessel morphology. AB - Intravascular ultrasound (IVUS) is an invasive imaging technique, characterized by the ability to depict the tomographic section of blood vessels' lumen and wall. Thus, it enables the performance of a unique in-vivo assessment of the luminal area and three-layer wall morphology, as well as atherosclerosis within the vessel. The typical composition of an IVUS image and the measurements of observed structures have been shown to correlate well with histology and angioscopy in several in vitro studies. The existing data on intravascular imaging of the aorta are scant. In this study, the preliminary results of in vitro IVUS examination of human descending aorta morphology are presented, with reference to the corresponding histological section. PMID- 12725504 TI - Multiple muscular variations in the neck region--case study. AB - During routine educational dissection studies, we encountered multiple muscular anomalies in a 25-year-old embalmed male cadaver. The muscular anomalies were observed on the right side. In accordance with their origin, insertion and innervation features, the abnormal muscles were considered sternocleidooccipital, sternomastoid and cleidomastoid muscles. The unilateral muscle variations were observed in both the superficial and deep layer. In the superficial layer, there were two muscles and one muscle bundle, while there was only one muscle in the deep layer. The first muscle in the superficial layer, known as sternocleidooccipital muscle, consisted of clavicular head and sternal head. The second muscle in the superficial layer was sternomastoid muscle. The muscle observed in the deep layer was cleidomastoid muscle. Additionally, a muscular bundle was observed between the sternocleidooccipital and sternomastoid muscles. A combination of these variations in the same case has not, to our knowledge, been previously described in the available literature. PMID- 12725506 TI - Pneumococcal disease in Australia: current status and future challenges. A report of the workshop held at the National Centre for Immunisation Research and Surveillance, 8-9 November, 2002. AB - In relation to surveillance, the predominant issue discussed was universal versus sentinel enhanced surveillance of IPD. In northern Australia, it will be important for enhanced surveillance to continue and to be as complete as possible. There are a number of reasons for this. First, the high incidence and high serotype diversity of IPD in Indigenous children in these areas has prompted the recommendation for boosters with 23vPPV to increase serotype coverage. This makes high quality, comprehensive surveillance essential for national policy. It is also important internationally as such as vaccine program has not been implemented anywhere else but is potentially applicable to other comparable populations. Secondly, the small absolute numbers of cases require data to be accumulated as comprehensively as possible. In relation to vaccine issues, both 23vPPV and 7vPCV policy are important. There was strong support from the meeting for the recent recommendation from the Australian Technical Advisory Group on Immunisation that both 23vPPV (for those over 65 years) and 7vPCV (for those less than 2 years) be publicly funded as universal programs. With respect to the current programs, there were important issues for Aboriginal and Torres Strait Islander people for both 23vPPV and 7vPCV. For 23vPPV, research is required into both the utility and frequency of boosters in adults as well as any potential role for 7vPCV in adults. Improving the identification of Aboriginal and Torres Strait Islander children is important, especially in urban areas. PMID- 12725505 TI - Australia's notifiable diseases status, 2001: annual report of the National Notifiable Diseases Surveillance System. AB - In 2001 there were 104,187 notifications of communicable diseases in Australia reported to the National Notifiable Diseases Surveillance System (NNDSS). The number of notifications in 2001 was an increase of 16 per cent of those reported in 2000 (89,740) and the largest annual total since the NNDSS commenced in 1991. In 2001, nine new diseases were added to the list of diseases reported to NNDSS and four diseases were removed. The new diseases were cryptosporidiosis, laboratory-confirmed influenza, invasive pneumococcal disease, Japanese encephalitis, Kunjin virus infection, Murray Valley encephalitis virus infection, anthrax, Australian bat lyssavirus, and other lyssaviruses (not elsewhere classified). Bloodborne virus infections remained the most frequently notified disease (29,057 reports, 27.9% of total), followed by sexually transmitted infections (27,647, 26.5%), gastrointestinal diseases (26,086, 25%), vaccine preventable diseases (13,030 (12.5%), vectorborne diseases (5,294, 5.1%), other bacterial infections (1,978, 1.9%), zoonotic infections (1,091, 1%) and four cases of quarantinable diseases. In 2001 there were increases in the number of notifications of incident hepatitis C, chlamydial infections, pertussis, Barmah Forest virus infection and ornithosis. There were decreases in the number of notifications of hepatitis A, Haemophilus influenzae type b infections, measles, rubella, Ross River virus infections and brucellosis. This report also summarises data on communicable diseases from other surveillance systems including the Laboratory Virology and Serology Reporting Scheme and sentinel general practitioner schemes. In addition, this report comments on other important developments in communicable disease control in Australia in 2001. PMID- 12725507 TI - OzFoodNet: Enhancing foodborne disease surveillance across Australia: quarterly report, July to September 2002. PMID- 12725508 TI - Mumps and rubella surveillance in Victoria, 1993 to 2000. AB - Despite improving childhood coverage of the measles-mumps-rubella vaccine (MMR) in Victoria during the 1990s, mumps and rubella notifications in age groups eligible for vaccination persisted. This study reviewed the mumps and rubella surveillance data from 1993 to 2000 with a specific focus on method of diagnosis. There were 474 notifications of mumps over the seven-year period (annual median 61, range 40 to 77) and 3,544 notifications of rubella (annual median 297, range 66 to 1,165). The highest notifications rates for mumps were consistently among the 1-4 and 5-9 year age groups, whereas there was a marked change in the age distribution of rubella notifications during this interval. A large rubella outbreak occurred in 1995 with 1,165 notifications; the highest notification rates were males aged 15-24 years, infants under one year of age (males and females), and those aged 5-14 years (males and females), respectively. The susceptibility of 5-24 year olds reflects historical changes to the Australian Standard Vaccination Schedule. Rubella notifications returned to baseline levels in 1998 with the highest notification rates in infants aged under one year, and children aged 1-4 years. For both mumps and rubella, the majority of notifications for all age groups were clinically diagnosed, and were most common in children. PMID- 12725510 TI - Uptake of influenza vaccine among Aboriginal and Torres Strait Island adults in north Queensland, 2002. AB - Since 1999, the Commonwealth has provided free annual influenza vaccine for all at-risk Aboriginal and Torres Strait Islander adults. The uptake of the vaccine in this population in north Queensland in 2002 was determined using the state wide computerised immunisation register. Although 59.4 per cent of Aboriginal and Torres Strait Islander adults aged 50 years or over were vaccinated, the uptake in this age group exceeded 80 per cent in only the Cape York and Torres Strait and North Peninsula Area Health Service Districts (HSDs). Assuming that a third of Aboriginal and Torres Strait Islander adults 15-49 years of age had a medical risk factor, it was estimated that 85 per cent of those at-risk were vaccinated. There was considerable variation between HSDs, ranging from 159 per cent uptake in the Torres Strait and North Peninsula Area Health Service District to 48.7 per cent in the Cairns HSD. PMID- 12725509 TI - Varicella surveillance: simpler than you think? PMID- 12725511 TI - Variability of larval identification characters of exotic Aedes albopictus (Skuse) intercepted in Darwin, Northern Territory. AB - There is no morphological identification key to satisfactorily distinguish between the larvae of Aedes (Stegomyia) species with special consideration to endemic and exotic species in Australia. Difficulty in differentiation between exotic Aedes (Stegomyia) albopictus (Skuse) and Aedes (Stegomyia) scutellaris Walker has been described previously. Aedes (Stegomyia) larvae were collected during an interception from an overseas vessel in Darwin, Northern Territory, and link bred. The adults were identified as Ae. albopictus. The larval skins and larvae were used to describe the variation in hair features of larval segment VII that are used to identify Ae. albopictus. The median hair number of hair 1-VII was three, whereas the description in Huang's identification key states four. The median of hair 2-VII was one, confirming Huang. However, the variability was higher than described by Huang and nearly half of the specimens showed different hair numbers on both sides. Individual specimens are therefore not clearly distinguished from other members of the Aedes (Stegomyia) scutellaris group. This paper also describes the detection, elimination and surveillance procedures following the interception. These were successful in preventing the establishment of exotic Ae. albopictus in the Northern Territory following the interception. PMID- 12725512 TI - Rainfall and vector mosquito numbers as risk indicators for mosquito-borne disease in central Australia. AB - There have been 5 confirmed cases of Murray Valley encephalitis virus (MVE) infection in the Alice Springs region during the high rainfall years of 1999/00 and 2000/01, compared with one case in the preceding 9 years. There also appeared to be an increased prevalence of Ross River virus (RR) infection in the Alice Springs and Tennant Creek regions associated with high rainfall. This paper presents an analysis of summer rainfall from 1990/91 to 2000/01, numbers of seroconversion of sentinel chickens to MVE, and RR cases in both regions. In Alice Springs where summer rainfall (December to February) and average vector numbers in the December to March period are closely correlated, the analysis also included mosquito vector numbers and MVE cases. Summer rainfall over 100 mm was significantly associated with sentinel chicken seroconversions to MVE. From December to March there was also a significant association of average vector numbers (> or = 300) with seroconversions in sentinel chickens following high summer rainfall. MVE appears enzootic in the Tennant Creek region and epizootic in the Alice Springs region. In Alice Springs during December to March, there was a significant association of RR cases with rainfall over 100 mm and with average vector numbers over 300. There was also a significant correlation of summer rainfall with RR cases in Tennant Creek. Summer rainfall is a new and good early indicator of high risk for both MVE and RR disease in the Alice Springs locality and RR in the Tennant Creek locality. Although similar relationships between rainfall and vector abundance, and disease incidence probably exist in other areas of central Australia, rainfall and vector abundance thresholds will probably vary according to local climatic and environmental conditions. PMID- 12725514 TI - Australian Sentinel Practice Research Network. PMID- 12725513 TI - Surveillance of viral pathogens in Australia: respiratory syncytial virus. PMID- 12725515 TI - Surveillance systems reported in CDI, 2003. PMID- 12725516 TI - Communicable diseases surveillance. Highlights for 4th quarter, 2002. AB - Communicable disease surveillance highlights report on data from various sources, including the National Notifiable Diseases Surveillance System (NNDSS) and several disease specific surveillance systems that provide regular reports to Communicable Diseases Intelligence. These national data collections are complemented by intelligence provided by State and Territory communicable disease epidemiologists and/or data managers. This additional information has enabled the reporting of more informative highlights each quarter. The NNDSS is conducted under the auspices of the Communicable Diseases Network Australia. NNDSS collates data on notifiable communicable diseases from State or Territory health departments. The Virology and Serology Laboratory Reporting Scheme (LabVISE) is a sentinel surveillance scheme which collates information on laboratory diagnosis of communicable diseases. In this report, data from the NNDSS are referred to as 'notifications' or 'cases', and those from ASPREN are referred to as 'consultations' or 'encounters' while data from the LabVISE scheme are referred to as 'laboratory reports'. PMID- 12725517 TI - Nuclear reprogramming and the redirection of cell fate in Xenopus. PMID- 12725518 TI - Somatic cell nuclear transfer. AB - Embryos produced by nuclear transfer from a patient's somatic cell offer one potential source of embryonic stem cells for treatment of human degenerative diseases. As with all of the approaches to such therapy, this has both strengths and weaknesses. The cells would be histocompatible with the patient's cells, be expected to have a normal life span, and in principle be a source of any other cell type. However, the time taken and the costs involved in the isolation of the appropriate cell population would probably prohibit large-scale application. Clones have been produced from the cells of adults of five species, but similar studies in at least five other species have produced early embryos, but not offspring. A variety of somatic cells have been used as successful nuclear donors. The present procedures have proved to be repeatable, but are very inefficient when typically between 1% and 4% of reconstructed embryos develop to adulthood. The inefficiency is the accumulated effect of failure at all stages of development. There may be differences between species and donor cell type in the precise pattern of loss. This outcome is assumed to reflect the inappropriate expression of a large number of genes whose lethal effect is exerted at different stages. Improvements in the efficiency may depend upon understanding those mechanisms in the early embryo that establish the precise chromatin structure that governs development. PMID- 12725519 TI - Cloned mice and embryonic stem cell lines generated from adult somatic cells by nuclear transfer. AB - Mice can now be cloned from cultured and noncultured adult-, fetus-, male-, or female-derived cells. Using the mouse as a model, research is moving towards a comprehensive description of clones generated by somatic cell nuclear transfer. In addition, embryonic stem (ES) cell lines can be generated from adult somatic cells via nuclear transfer (ntES cells). ntES cells contribute to an extensive variety of cell types including neurons in vitro and germ cells in vivo. Recent advances in mouse cloning are reported to illustrate its strengths and promise in the study of mammalian biology and biomedicine. PMID- 12725520 TI - Developmental origins of hematopoietic stem cells. AB - Hematopoietic stem cells (HSCs) are at the foundation of the hematopoietic hierarchy and give rise to all blood lineages in the adult organism. A thorough understanding of the molecular, cellular, and developmental biology of HSCs is of fundamental importance, but is also clinically relevant for the advancement of cell replacement therapies and transplantation protocols in blood-related genetic disease and leukemias. While the major anatomical sites of hematopoiesis change during ontogeny, it was long believed that the developmental origin of the adult mammalian hematopoietic system was the yolk sac. However, current studies have shown that the first adult-type HSCs are autonomously generated in the intrabody portion of the mouse embryo, the aorta-gonads-mesonephros (AGM) region, and sublocalize to the dorsal aorta. HSCs are also found in the other large embryonic vessels, the vitelline and umbilical arteries. The intraluminal hematopoietic clusters along these vessels, together with the role of the Runx1 transcription factor in cluster and HSC formation and the HSC/endothelial/mesenchymal Runxl expression pattern, strongly suggest a vascular endothelial/mesenchymal origin for the first HSCs. Moreover, a transgenic mouse line expressing the GFP marker under the control of the Sca-1 transcriptional regulatory elements (GFP expression marks all HSCs) shows a clear localization of GFP-expressing cells to the endothelial cell layer of the dorsal aorta. Thus, highly enriched GFP positive AGM HSCs will serve as a basis for the future examination of the cellular and molecular factors involved in the induction and expansion of adult HSCs. PMID- 12725521 TI - RhoGTPases and their role in cancer. AB - The RhoGTPases constitute a large family of small GTP binding proteins that police many sensitive crossroads in diverse cellular functions. Therefore, it would not be surprising if aberrant RhoGTPase function led to perturbed growth and differentiation, including carcinogenesis and cancer progression. The evidence for a causative connection between RhoGTPases and primary human cancers is still weak but there are increasing findings to support this link. An appreciation of this connection is timely and important to alert readers to the possibilities of new disease mechanisms and new ways to diagnose and treat cancer. PMID- 12725522 TI - Analyzing tumor suppressor activities in the murine small intestine. AB - We have used mice deficient in a series of genes with either known or potential tumor suppressive activity to determine the phenotype of loss of function of these genes in the mouse. We have tested a series of endpoints that derive from the hypothesis that loss of an apoptotic program would be predicted to fail to delete cells carrying DNA damage, that this would lead to increased clonogenic survival and thereby to an increased mutation burden and tumor predisposition. For p53 deficiency, we show that loss of the apoptotic program does not translate into an increase in spontaneous mutation rate. However, p53 deficiency can lead to increased clonogenic survival, although this is highly damage-type dependent. Furthermore, p53 deficiency weakly accelerates tumorigenesis associated with inactivation of the adenomatous polyposis coli gene, Apc. We have also analyzed mice mutant for the mismatch repair genes Msh2, Mlh1, and Pms2, describing circumstances in which all of these strains show defective apoptosis, increased clonogenic survival, and increased mutation rate. However, these effects are highly drug-type dependent and the pattern of dependency argues strongly that mutation rate increases as a direct result from loss of the apoptotic program. We have also identified a new role for p53 by intercrossing the p53 and Msh2 mutants, so demonstrating that heterozygosity for p53 accelerates microsatellite instability. Finally, we have analyzed mice mutant for Mbd4 and show that this gene functions in vivo as a tumor suppressor. PMID- 12725523 TI - The developmental program of murine erythroleukemia cells. AB - Hematopoietic stem cells (HSCs) or early progenitors respond to external stimuli in bone marrow and differentiate into cell-restricted lineages of blood cells of limited life span. In leukemias, however, early hematopoietic progenitors self renew themselves, fail to respond to differentiation signals, and do not undergo programmed cell death (apoptosis). The basic mechanisms of differentiation and apoptosis of leukemia cells have been the long-term objective of our work. By exploiting widely studied murine and human leukemic cell systems as models of hematopoietic cell differentiation, we explored the mechanisms by which pharmaceutical agents initiate differentiation in leukemic systems. In this article, we present the developmental program of MEL cells with emphasis given on the role of commitment to terminal maturation. Commitment is initiated via inducer-receptor-mediated processes and leads to discrete patterns of expression of several genes that contribute to growth arrest at the G1 phase, expression of differentiated phenotype, and differentiation-dependent apoptosis (DDA). Overall, MEL erythroid cell differentiation represents a developmental program with a highly coordinated set of processes that is "triggered" by an inducer and functions via a network of genes and proteins interacting with each other harmonically to give birth to lineage-restricted phenotype. PMID- 12725524 TI - Small molecule regulation of phosphatase-dependent cell signaling pathways. AB - Small molecules provide exceptionally useful tools for probing signaling targets relevant for cancer and stem cell differentiation. In contrast to genetic approaches, the application of small molecules generally offers a graded and reversible disruption of a particular pathway. The vast array of theoretical chemical entities that exist in the chemical universe are now becoming available through the production and distribution of chemical libraries generated by both traditional and combinatorial methods, which are suitable for pharmacological use. Convenient and inexpensive cell-free and cell-based assays can be used to identify chemicals that exhibit desirable antisignaling properties. We illustrate a model of how agents targeted against signaling macromolecules involved in cancer, namely dual specificity protein phosphatases, can be identified. PMID- 12725525 TI - Isolation and characterization of normal adult human epithelial pluripotent stem cells. AB - Both reproductive and therapeutic cloning of human stem cells have been made possible with recent technological advances in the isolation of embryonic stem cells and of pluripotent stem cells from adult tissues. We have isolated normal human kidney and human breast epithelial stem cells, as well as having characterized "immortalized" cells from human neuronal and human pancreatic tissue (Trosko et al., Methods 20:245-264, 2000). The isolation was motivated by the stem cell theory of carcinogenesis. Based on the assumption that stem cells would not express connexin genes, nor have functional gap junctional intercellular communication (GJIC), we have demonstrated that the human kidney, breast, neuronal, and pancreatic stem cells can divide either symmetrically or asymmetrically, depending on whether they are grown in microenvironmental conditions that suppress GJIC (the undifferentiated, proliferative state) or induce GJIC (the differentiated state). Normal breast epithelial stem cells appear to be intrinsically "immortal" until induced to express GJIC, at which time, with appropriate substrate and microenvironmental nutrients, they can form three-dimensional "organoids." expressing markers associated with the mature mammary tissue and forming a physical structure very similar to the in vivo budding, ductal structures. The breast stem cells can be prevented from "mortalizing" and can be converted to neoplastic cells, which maintain many phenotypes of the stem cells. PMID- 12725526 TI - Functional cloning of IGFBP-3 from human microvascular endothelial cells reveals its novel role in promoting proliferation of primitive CD34+CD38- hematopoietic cells in vitro. AB - Ex vivo expansion of hematopoietic stem cells (HSCs) has been investigated as a means of enhancing engraftment of transplantation therapies, but current ex vivo expansion methods typically result in a loss of functional stem cell activity. Factors that can selectively expand human HSCs remain elusive. Recently we have isolated three functionally distinct human brain microvascular endothelial cells (HBMVECs) that differ greatly in their ability to support in vitro proliferation of human umbilical cord blood (UBC) CD34+CD38-cells. Using these distinct HBMVEC populations, we have devised a cell-based functional cloning assay to identify a molecule(s) capable of facilitating expansion of HSCs in vitro. A gene encoded for IGFBP-3 (insulin-like growth factor binding protein-3) has been identified. IGFBP-3 mRNA and protein are differentially expressed in distinct HBMVEC populations. In vitro cell proliferation assay and CD34+CD38- immunophenotype analysis showed that the addition of an exogenous IGFBP-3 to cultures of purified CD34+/-CD38-Lin- cells (CD2/CD3/CD14/CD16/CD19/CD24/CD56/CD66b/GlyA depleted) enhanced proliferation of primitive hematopoietic cells with CD34+CD38- phenotype, suggesting that IGFBP-3 is capable of expanding primitive human blood cells. These expanded primitive blood cells were illustrated to maintain ability to generate functional progenitors. IGFBP-3 belongs to a family of high-affinity IGFBPs, which binds to IGFs and modulates their actions. IGFBP-3 appears to have intrinsic bioactivity that is independent of IGF binding. We are currently exploring the underlying mechanism by which IGFBP-3 modulates proliferation of primitive hematopoietic cells, and the potential of IGFBP-3 to expand pluripotent human repopulating cells capable of hematopoietic reconstitution of irradiated NOD/SCID recipients. PMID- 12725527 TI - Insights into the role of DNA methylation in murine embryonic stem cells using a modified tetracycline-inducible gene expression system. AB - Murine embryonic stem (ES) cells are pluripotent cells derived from the inner cell mass of the mouse blastocyst. Their ability to differentiate into diverse cell types in vitro has served well as a model of early embryonic developmental processes. A powerful strategy for studying the role of key genes during this time is to be able to regulate their expression in a dose-dependent manner. Here, we outline our experience of applying the "tetracycline-inducible system" to study gene function during ES cell differentiation and describe a number of key modifications designed to overcome the central problem of transgene silencing. Finally, we demonstrate how this approach can yield insights into the role of DNA methylation during ES cell differentiation by studying the function of two genes: DNMT3A, involved in the de novo methylation of cytosine bases, and MBD2b, a methyl-dependent co-repressor and putative cytosine demethylase. PMID- 12725528 TI - Emerging interest in the kallikrein gene family for understanding and diagnosing cancer. AB - Kallikreins are proteolytic enzymes that constitute a subfamily of serine proteases. Novel kallikrein genes were cloned recently, and it was shown that the human kallikrein family contains 15 genes tandemly aligned on chromosomal locus 19q13.3-q13.4. Based on their altered expression in tumor cells, kallikreins may be involved in the pathogenesis and/or progression of cancer. Evidence is presented that certain kallikreins may be exploited as diagnostic cancer biomarkers. Although the function(s) of novel kallikreins is currently unknown, increasing evidence suggests that kallikreins may participate in regulatory enzymatic cascade(s). Elucidation of the function of novel kallikreins largely depends on the availability of active recombinant proteins. Here, the zymogen for kallikrein 13 was overexpressed in Pichia pastoris and biochemically characterized. It was shown that the kallikrein 13 zymogen displays intrinsic catalytic activity leading to autoactivation. A clipped form of kallikrein 13 was identified, indicating autocatalytic cleavage at the internal bond R114-S115. Mature kallikrein 13 displays trypsin-like activity with restricted specificity on synthetic and protein substrates. Combinatorial P1-Lys libraries of tetrapeptide fluorogenic substrates were synthesized and used for the profiling of the P2 specificity of selected kallikreins. Interestingly, it was shown that human kallikrein 13, similarly to PSA, could specifically cleave human plasminogen to generate angiostatin-like fragments, suggesting that specific kallikreins may have antiangiogenic actions. An understanding of the physiology of human kallikreins is emerging with potential clinical applications. PMID- 12725529 TI - Identification of keratinocyte proteins that mark subsets of cells in the epidermal stratum basale: comparisons with the intestinal epithelium. AB - Rapid renewing epithelia such as the epidermis and the intestinal epithelium are maintained by proliferation of undifferentiated stem cells located at specific locations. Recent experiments indicate that stem cells from adult organs might be able to populate tissues other than their tissue of origin. Such findings open the possibility that adult stem cells from different tissues might share common markers. We investigated this by two different approaches. In a first approach we compared the expression profiles from epidermal and intestinal epithelial cells at various stages of differentiation. We found that 108 of 1,176 genes analyzed were expressed above background in either keratinocytes or enterocytes and, among these, only 16 genes were expressed in both cell types. Of these 16 genes expressed in both cell types, only five displayed the same shift in expression level during cellular differentiation. Interestingly, all five genes were downregulated during cellular differentiation and represented ubiquitously expressed genes. In the second approach we analyzed the expression of the CC chemokine receptor 6 (CCR6), which we have recently identified as an early differentiation marker of epidermal cells, in the intestine. This analysis demonstrates that the CCR6 protein is found in enterocytes at later stages of differentiation. PMID- 12725531 TI - A new method for determining the status of p53 in tumor cell lines of different origin. AB - The tumor suppressor p53 is the most frequently mutated gene in human tumors. In response to DNA damage, aberrant growth signals, or chemotherapeutic drugs, p53 is stabilized and induces apoptosis and/or cell cycle arrest. While the mechanisms of p53-dependent apoptosis are not well understood, p53-dependent cycle arrest is primary mediated by the CDK inhibitor p21. p53 is a transcriptional activator and it is not surprising that a majority of p53 mutations occur in the core DNA binding domain and affect DNA binding and transactivation of p53 targets in tumors. We used the capability of p53 to activate transcription for developing a new assay that permits rapid determination of the status of p53 in cancer cell lines of different origin. Our strategy involved using a retrovirus containing a p53-regulated lacZ reporter gene that was introduced into colon and breast tumor cell lines to determine p53 status. Simple staining for beta-galactosidase allowed us to confirm that the colon cancer cell lines LIM1215 and HCT116, as well as the breast cancer cell line MCF7. have wild-type p53, and the colon cancer cell line Caco-2 as well as breast cancer cell lines MDA-MB-435 and MDA-MB-231 have mutant p53. This method may be applied to novel cell lines of any origin with unknown status of p53. PMID- 12725530 TI - Cisplatin resistance in an ovarian carcinoma is associated with a defect in programmed cell death control through XIAP regulation. AB - Chemoresistance is a major impediment to the successful treatment of cancer. It involves various mechanisms, including defects in the apoptosis program that is induced by anticancer drugs. To further explore the mechanisms underlying the development of chemoresistance in ovarian carcinoma after cisplatin (CDDP) treatment, we compared the effect of CDDP on expression of X-linked inhibitor of apoptosis protein (XIAP), a direct inhibitor of caspase-3, -7, and -9, Fas, Fas ligand (Fas-L), and pro- and antiapoptotic proteins in a CDDP-sensitive human ovarian carcinoma cell line (2008) and its CDDP-resistant subclone (2008C13). In this article, we show that cisplatin treatment led to a differential expression of distinct apoptotic targets in the CDDP-sensitive cell line (2008) and its CDDP resistant subclone (2008C13). The acquisition of cisplatin resistance was associated with the ability of the treated cells to enhanced expression of XIAP, whereas the death inducer Fas-L was abrogated in 2008C13 following treatment with CDDP. However, the CDDP-sensitive cells failed to activate XIAP but increased Fas L expression, indicating that distinct regulatory mechanisms are operative. These findings suggest that the expression of XIAP and downregulation of Fas-L are linked to chemoresistance in ovarian carcinoma cells and may represent one of the potential antiapoptotic mechanisms involved during this process. PMID- 12725532 TI - Brk, Srm, Frk, and Src42A form a distinct family of intracellular Src-like tyrosine kinases. AB - The tyrosine kinases Brk/PTK6/Sik, Srm, Frk/Rak/Gtk/Iyk/Bsk, and Src42A/Dsrc41 have a low degree of sequence homology to other known kinases, including one another. We show here that the exon structure of these kinases, which we will call the Brk family, is highly conserved and distinct from each of the major families of intracellular kinases containing SH3, SH2, and tyrosine kinase domains, including c-Src and Fyn. Brk/Sik and Srm are 1.1 kb apart on human chromosome 20q13.3 and likely are the result of duplication in cis. Several Brk family kinases have an inhibitory effect on Ras pathway signaling from receptor tyrosine kinases. Members of this family can act either in the membrane or at the nucleus, and may change localization patterns depending on external stimuli. Brk has been shown to phosphorylate two proteins in vivo: Sam68. a substrate for Src in mitosis that can substitute for Rev in nuclear export of RNAs; and BKS, a novel adaptor molecule. Brk also functions as a rapid downstream signaling intermediate following calcium-induced differentiation in keratinocytes. It is possible that Brk family kinases may share common functions and interaction partners, which remain for the most part unexamined. PMID- 12725533 TI - Structural genomics of lipid signaling domains. AB - Signaling domains have been identified by the analysis of data derived from biochemical studies, molecular cloning, or genetic studies. With the availability of genomic information from many organisms and the improved sensitivity in homology detection techniques, many new domains are being identified. In an attempt to understand biochemical and biological function of these domains, we have started a small-scale structural genomics, or structural biology with genomic approach. Two examples from our recent work are steroidogenic acute regulatory protein (StAR)-related lipid-transfer (START) domain and inositol polyphosphate 5-phosphatase catalytic (IPP5C) domain. Crystal structure of human MLN64-START domain revealed a hollowed-out protein containing a hydrophobic tunnel just large enough to bind one molecule of cholesterol and completely exclude it from solvent. This structure suggests that the START domain is a classical type of lipid transporter. On the contrary, the function of IPP5C domain has been extensively studied for a long time, but its catalytic mechanism, positional selectivity, and diverse substrate specificity remained mysterious due to the unavailability of three-dimensional structure. With the structural genomic approach, the first structure of IPP5C domain was solved from a S. pombe protein that is now known as SPsynaptojanin and the structure gave us answers to some of these questions. PMID- 12725534 TI - Differentially expressed downstream genes in cells with normal or mutated p53. AB - Mutations in p53 gene could lead to loss of function, negative complementation, or to gain-of-oncogenic functions, thus leading to the increase of tumorigenicity and invasiveness of cancer. This study focused on cancer-related p53 mutants, including A138T, C141Y, RI58L, G245C, and R248Q. Using a modified differential display technique, response profiles of plasmid-expressed wild-type as well as mutated p53, in comparison to p53-null cells, are being established. These profiles can help our understanding of p53 involvement in cancer, leading to accurate diagnosis, treatment, and prognostic analysis by: 1) comprehensive knowledge of p53 network gene expression profiles; 2) finding the most significant gene expression profiles of p53 mutants; 3) revealing genes that only respond to p53 mutants (gain of function). Our results showed significant differences in the expression patterns among p53-null. wild-type p53, and p53 mutants A138T, C141Y, R158L, G245C, and R248Q samples. We also report here the first found p53 mutant-triggered alternative splicing. PMID- 12725535 TI - Oncoselective parvoviral vector-mediated gene therapy of cancer. AB - We replaced capsid genes by reporter genes and assessed expression in different types of human cancer cells and their normal counterparts, either at the level of whole cell population (CAT ELISA) or at the single cell level [FACS analysis of green fluorescent protein (GFP)]. CAT expression was substantial (up to 10,000 times background) in all infected tumor cells, despite variations according to the cell types. In contrast, no gene expression was detected in similarly infected normal cells (with t he exception of an expression slightly above background in fibroblasts). FACS analysis of GFP expression revealed that most tumor cells expressed high level of GFP while no GFP-positive normal cells could be detected with the exception of very few (less than 0.1%) human fibroblast cells expressing high level of GFP. We also replace capsid genes by genes coding for the costimulatory molecules B7-1 and B7-2 and show that, upon infection with B7 recombinant virions, only tumor cells display the costimulatory molecules and their immunogenicity was increased without any effect on normal cells. Using a recombinant minute virus of mice (MVM) containing the herpes simplex thymidine kinase gene, we could get efficient killing of most tumor cell types in the presence of ganciclovir. without affecting normal proliferating cells. The prospects and limitations of these different strategies are discussed. PMID- 12725536 TI - Control of hematopoietic stem/progenitor cell fate by transforming growth factor beta. AB - A major obstacle to the use of adult somatic stem cells for cell therapy is our current inability to fully exploit stem cell self-renewal properties. The challenge is to obtain defined culture systems where cycling of primitive stem/progenitor cells is stimulated, while their differentiation and senescence are prevented. The cytokine transforming growth factor-beta1 (TGF-beta1) appears as a potential regulator of hematopoietic stem/ progenitor cell self-renewal, as it participates in the control of cell proliferation, survival/apoptosis, and cell immaturity/differentiation. TGF-beta1 acts via a complex regulatory network involving intracellular signaling molecules and cell surface receptors. According to the High Proliferative Potential-Quiescent (HPP-Q) cell working model that we introduced previously, TGF-beta1 maintains primitive hematopoietic stem/progenitor cells in a quiescent or slow cycling state, in part by downmodulating the cell surface expression of mitogenic cytokine receptors, thus preventing cells from responding rapidly to a mitogenic signal. We have established that this modulation concerns the tyrosine kinase receptors KIT and FLT3, and the IL-6 receptor (IL-6R), three important cytokine receptors controlling early human hematopoietic stem/progenitor cell development. In this article. we show a similar modulation by TGF-beta1 of a fourth receptor: the TPO receptor (MPL). As a consequence, TGF-beta1 decreased the cell cycle entry of stem/progenitor cells stimulated by the respective ligands of these receptors, the cytokines SF, FL, IL-6, and TPO, whereas neutralization of TGF-beta1 increased the cell responsiveness to these mitogenic cytokines. Other aspects of the function of TGF-beta1 in the regulation of early hematopoiesis (i.e., the control of stem/progenitor cell survival and immaturity) are reviewed in the discussion. PMID- 12725537 TI - Intended plans for breastfeeding duration: a simple tool to predict breastfeeding outcome. AB - Prenatal intended duration of breastfeeding has been shown to be a powerful predictor of breastfeeding initiation and duration. This provides a tool to establish the risks for short duration of breastfeeding. CONCLUSION: It is necessary to identify the factors that influence mothers' plans, and to find means to intervene effectively. PMID- 12725538 TI - Beneficial effects of postnatal skin-to-skin contact. AB - In a randomized trial, Bystrova et al. examined the traditional neonatal practice of swaddling. They demonstrated that extended skin-to-skin contact with the mother is the most effective way to maintain the infant's temperature and decrease the "stress of being born". CONCLUSION: Only through research such as that reported by Bystrova et al. can the advantages or disadvantages of routinely accepted perinatal practices be evaluated. PMID- 12725539 TI - Effects of neonatal intensive care on girls' and boys' language development. AB - According to Jennische and Sedin, very premature boys are less affected in their speech and language development than a matched group of girls. This is discussed from a general linguistic perspective. CONCLUSION: It is speculated that perinatal factors, timing and ambient aspects may impact on boys' advantage over girls. PMID- 12725540 TI - Secular changes in body height and weight in children and adolescents in Poznan, Poland, between 1880 and 2000. AB - AIM: A secular trend in body height and weight is well documented. The first observations concerning this phenomenon in Poland were made at the end of the 19th century. The aim of this study was to assess changes in body height and weight during the 20th century, with special emphasis on the last decade. METHODS: The results of body height and weight measurements obtained in eight subsequent surveys (1880-1886, 1922-1927, 1946-1950, 1960-1961, 1970-1971, 1980 1981, 1990-1991 and 1999-2000) were included in the analysis. Mean values were compared and differences between the surveys were analysed. RESULTS: In general, in the 20th century, children grew taller and heavier and reached final body height and weight more rapidly. The biggest differences in body height and weight in the 20th century, observed at growth spurt, were about 17 cm and 11 kg, respectively, for boys, and 13 cm and 13 kg for girls. The magnitude of secular changes in body height and weight in the 20th century was not stable. There were periods of increased and decreased intensity of acceleration of physical development (the 1950s and 1970s, and the 1960s and 1980s, respectively), as well as a period of deceleration (the 1940s). In the last decade, the tendency has been towards deceleration in most age groups studied. CONCLUSION: The acceleration of physical development in children in Poznan has now ceased. PMID- 12725541 TI - Plasma lactate and plasma volume recovery in adults and children following high intensity exercises. AB - AIM: To compare plasma lactate concentration recovery kinetics when measured and corrected for variations in plasma volume between children and adults. METHODS: Nine boys (11.3 +/- 1.1 y) and 8 men (21.9 +/- 1.9 y) performed a maximal and a supramaximal exercise until exhaustion. Plasma lactate concentrations, haemoglobin and haematocrit were measured at rest, immediately on completion of exercise and after the 2nd, 5th, 12th and 30th minute of recovery. The plasma lactate concentrations and the rate of recovery were corrected for variations in plasma volume. RESULTS: The maximal decreases in plasma volume were significantly higher in adults than in children for maximal exercise (-18.7 +/- 2.6% vs -14.5 +/- 3.2%; p < 0.05), but similar for the supramaximal exercise (-16.9 +/- 3.4% vs -15.2 +/- 3.4%). During recovery, measured and corrected plasma lactate concentrations were significantly higher in adults. The rate of plasma lactate recovery was higher in adults for maximal exercise only. The same results were obtained when the rates of plasma lactate decrease were calculated from corrected plasma lactate concentrations. CONCLUSION: The correction of the plasma lactate concentrations for variations in plasma volume did not influence the comparison of the concentrations obtained in adults and children, or their rate of recovery. PMID- 12725542 TI - Treatment of urinary tract infection with gentamicin once or three times daily. AB - AIM: To examine the safety and efficacy of once-daily (OD) gentamicin treatment compared with conventional 8-hourly dosing (TDS) for urinary tract infection (UTI). METHODS: This was a prospective, randomized, controlled trial of children 1 mo to 13 y of age with presumed UTI. Children were randomly assigned to OD gentamicin 5 mg kg(-1) d(-1) or TDS gentamicin 6 mg kg(-1) d(-1) divided 8 hourly. Microbiological efficacy, nephrotoxicity, ototoxicity and renal scarring were assessed at the end of treatment. RESULTS: 210 patients with presumed UTI were recruited, of whom 172 were analysable (OD 84, TDS 88). The median age was 7 mo, 50% were male and 74% (n = 127) of patients had pyelonephritis. The majority of infections were due to Escherichia coli (n = 153, 89%), of which 9 (5.2%) were bacteraemic. Comparing the two groups, there was no significant difference in age, gender, duration of fever before admission, pyuria, nitrite positivity or initial total white blood cell count. All patients had negative urine cultures after 2-3 d of treatment, demonstrating 100% microbiological efficacy. There was no difference between the two groups in terms of ototoxicity, nephrotoxicity, duration of gentamicin treatment or time to fever defervescence. CONCLUSION: OD gentamicin is as efficacious as TDS gentamicin in the treatment of UTI in children, with no difference in ototoxicity and nephrotoxicity. PMID- 12725543 TI - Ultrasonic tissue characterization of the myocardium in anorexia nervosa. AB - AIM: To determine the systodiastolic variations in the integrated backscatter (IBS) signal of the myocardium in patients with anorexia nervosa. METHODS: 25 young women (aged 22.4 +/- 4.3 y) with overt anorexia nervosa, compared with 25 age-matched thin and 25 age-matched control women with body mass index >20 kg m( 2), underwent either conventional two-dimensional echocardiography or analysis of IBS cyclic variations. RESULTS: Compared with thin and control subjects, anorectic patients showed reduced left ventricular mass (LVM: 82.9 +/- 17.1 vs 119.9 +/- 13.8 and vs 126.12 +/- 16.4 g, p < 0.0001; LVM indexed 21.4 +/- 3.3 vs 29.4 +/- 2.5 and vs 31.2 +/- 3.1 g m(-2.7), p < 0.0001), and IBS cyclic variations (septum: -0.49 +/- 2.18 vs 6.86 +/- 1.3 and vs 6.61 +/- 1.74 dB p < 0.0001; posterior wall: 2.77 +/- 2.12 vs 7.15 +/- 2.12 and vs 7.48 +/- 2.23 dB, p < 0.01). CONCLUSION: Anorexia nervosa is associated with a significant reduction in the cyclic variation in IBS, which is also related to left ventricular hypotrophy. Ultrasonic tissue characterization could give an objective approach for the detection of myocardial structural properties and represent a preclinical index of myocardial dysfunction in anorexia nervosa. PMID- 12725544 TI - Posterior ocular malformations in children: somatic, neuroradiological and cognitive aspects. AB - AIM: To investigate associated neurological, endocrinological and cognitive dysfunctions in children with visual impairment caused by optic nerve hypoplasia or optic nerve/fundus coloboma and/or microphthalmus. METHODS: Forty children born between 1990 and 1998 were assessed by neurological examination, re evaluation of neuroradiological investigations, review of medical records and examination of cognitive levels. RESULTS: Neurological dysfunctions (epilepsy and/or motor impairment) were found in 13/28 children with optic nerve hypoplasia and in 3/12 children with coloboma/microphthalmus. The optic pathways were thin in 22/24, an abnormal posterior pituitary gland and/or thin infundibulum was found in 16/22 and absence of septum pellucidum in 14/27 children with optic nerve hypoplasia. Other types of cerebral abnormalities occurred in 9/26 children. Among children with coloboma/microphthalmus, the optic pathways were thin in 4/8 children but none had pituitary or cerebral midline abnormalities. Sixteen children with optic nerve hypoplasia were receiving hormone substitution but none of the children with coloboma/microphthalmus. Thirteen of the 16 children with optic nerve hypoplasia and with an abnormal pituitary region had endocrinological deficiencies. Mental retardation was found in 9/24 of the children with optic nerve hypoplasia and in 5/10 of the children with coloboma/microphthalmus. CONCLUSION: Endocrinological. neurological and neuroradiological defects seem more common in children with optic nerve hypoplasia than in children with coloboma. A pituitary region that appears abnormal in magnetic resonance imaging seems to predict endocrinological deficits in children with optic nerve hypoplasia. PMID- 12725545 TI - How reliable is axillary temperature measurement? AB - AIM: To assess whether axillary temperature measurements reliably reflect oral/rectal temperature measurements. METHODS: This observational study compared paired axillary-rectal and axillary-oral temperatures in a general paediatric ward with the participation of 225 children aged < or = 4 y and 112 children aged between 4 and 14 y. RESULTS: Changes in oral/rectal and axillary temperatures correlated significantly (p < 0.0001). However, axillary temperature measurements were significantly lower than both oral (mean -0.56 degrees C, SD 0.76 degrees C) and rectal measurements (0.38 degrees C; SD 0.76 degrees C). Ninety-five percent of axillary measurements fell within a 2.5-3 degrees C range around respective paired oral/rectal measurements. The mean difference increased with increasing temperature, and was 0.4 degrees C at low body temperatures, and over 1 degree C with a fever of 39 degrees C. Neither seasonal fluctuations nor the amount of clothing worn influenced this difference. CONCLUSION: Axillary temperatures in young children do not reliably reflect oral/rectal temperatures and should therefore be interpreted with caution. PMID- 12725546 TI - Lung lavage using high-frequency jet ventilation in rabbits with meconium aspiration. AB - AIM: To determine the efficacy of the expulsion effect of high-frequency jet ventilation (HFJV) on meconium clearance from the airways in comparison with conventional suctioning in adult rabbits with meconium aspiration. METHODS: Experiments were carried out on tracheotomized, anaesthetized and paralysed adult rabbits. A suspension of human meconium in saline (25 mg ml(-1), 4 ml kg(-1)) was instilled into the tracheal cannula. When respiratory failure developed, saline lavage (10 ml kg(-1) in 3 portions) was performed during conventional ventilation or by means of the inpulsion and expulsion regime of HFJV. Animals were further ventilated for 2 h with either conventional ventilation or HFJV. RESULTS: There was no significant difference between groups in the amount of meconium recovered by lavage. Compared to conventional ventilation, the application of HFJV enhanced the elimination of carbon dioxide, increased lung compliance and diminished right to-left shunts after 30 min of ventilatory treatment. Oxygenation also improved during HFJV, although this was not a consistent finding during the ventilation period. CONCLUSION: HFJV improved gas exchange, lung compliance and reduced right to-left pulmonary shunts, but saline lung lavage by HFJV was not found to be more efficient than lavage during conventional ventilation in rabbits with meconium aspiration. PMID- 12725547 TI - Skin-to-skin contact may reduce negative consequences of "the stress of being born": a study on temperature in newborn infants, subjected to different ward routines in St. Petersburg. AB - AIM: To evaluate how different delivery-ward routines influence temperature in newborn infants. METHODS: A total of 176 newborn mother-infant pairs were included in a randomized study. The babies were kept skin-to-skin on the mother's chest (Skin-to-skin group), held in their mother's arms, being either swaddled or clothed (Mother's arms group), or kept in a cot in the nursery, being either swaddled or clothed (Nursery group). Temperature was measured in the axilla, on the thigh, back and foot at 15-min intervals at from 30 to 120 min after birth. RESULTS: During this time period the axilla, back and thigh temperatures rose significantly in all the treatment groups. The foot temperature displayed a significant fall in the babies in the Nursery group and this decrease was greatest in the swaddled babies. In contrast, foot temperature rose in the babies in the Mother's arms group and in particular in babies in the Skin-to-skin group. Foot temperature remained high in the Skin-to-skin group, whereas the low temperature observed in the Nursery group gradually increased and two days after birth the difference was no longer significant. CONCLUSION: The results show that delivery-ward routines influence skin temperature in infants in the postnatal period. Allowing mother and baby the ward routine of skin-to-skin contact after birth may be a "natural way" of reversing stress-related effects on circulation induced during labour. PMID- 12725548 TI - Transcutaneous bilirubinometry during and after phototherapy. AB - AIM: To evaluate the reliability of transcutaneous bilirubinometry (TcB) during and after phototherapy. METHODS: TcB was performed on the forehead and chest of infants with neonatal jaundice when capillary blood was sampled for bilirubin determination in a control group of 240 neonates. In a second group of 70 neonates exposed to phototherapy the same procedure was performed after at least 24 h of exposure: on the forehead, TcB was done in the centre of the unexposed area and also on the adjacent exposed area, and the exposed chest. During the post-phototherapy period, TcB was again done during the first and second days, at least 18-24 h after cessation of phototherapy. The results were then statistically evaluated and regression curves were plotted. RESULTS: A close correlation between TcB values and bilirubin levels was observed in the control group. In the phototherapy group, a correlation was also found between the TcB and the bilirubin values, but this correlation was significantly poorer than that of the controls; the correlation for the covered part of the forehead was significantly better than that of the exposed part but still poorer than that of the controls, though the difference was no longer significant. Skin colour recovered during the post-phototherapy period and correlation was better than that during exposure and no longer significantly different from that of the controls. CONCLUSION: Through its bleaching effect on the skin, phototherapy affects the correlation between TcB and the bilirubin values, but does not totally eliminate it. The unexposed parts show a better correlation, though this was still poorer than that of the controls. Recovery of skin colour occurred within 18-24 h after cessation of exposure. PMID- 12725549 TI - Implementation of a nation-wide automated auditory brainstem response hearing screening programme in neonatal intensive care units. AB - AIM: As part of a future national neonatal hearing screening programme in The Netherlands, automated auditory brainstem response (AABR) hearing screening was implemented in seven neonatal intensive care units (NICUs). The objective was to evaluate key outcomes of this programme: participation rate, first stage success rate, pass/referral rates, rescreening compliance, diagnostic referral rates, age of first diagnostic evaluation and prevalence of congenital hearing loss (CHL). METHODS: This prospective cohort study collected data on 2513 survivors. NICU graduates with one or more risk factors according to the Joint Committee on Infant Hearing were included in a two-stage AABR hearing screening programme. Conventional ABR was used to establish a diagnosis of CHL. RESULTS: A total of 2513 newborns enrolled in the programme with a median gestational age of 31.6 (range 24-43) wk and a median birthweight of 1450 (range 510-4820) g. In 25 (1%) cases parents refused screening. Four out of 2513 newborns were initially lost; 2484 newborns have been tested initially. A final 98% participation rate (2465/2513) was obtained for the whole programme. After a median postmenstrual age at the first test of 33.7 (range 27-54) wk, a pass rate of 2284/2484 (92%) resulted at the first stage. The rescreening compliance after the first test was 92% (184/200). A referral rate for diagnostic ABR of 3.1% (77/2484) resulted. Of the 77 referrals 14 (18.2%) had normal screening thresholds, 15 (19.5%) had unilateral CHL and 48 (62.3%) had bilateral CHL. The prevalence of unilateral CHL was 0.6% (15/2484) and of bilateral CHL 1.9% (48/2484). CONCLUSION: A financially supported two-stage AABR hearing screening programme can be successfully incorporated in NICU centres and detects a high prevalence of CHL in NICU graduates. Neonatal hearing screening should be part of standard clinical practice in all NICU infants. PMID- 12725550 TI - Effect of perinatal asphyxia on thyroid-stimulating hormone and thyroid hormone levels. AB - AIM: To compare serum concentrations of thyroid hormones--T4, T3, free T4 (FT4) and reverse T3 (rT3)--and thyroid-stimulating hormone (TSH) found in the umbilical cord blood of term newborns with and without asphyxia and those found in their arterial blood collected between 18 and 24 h after birth. A further aim of the study was to assess the association between severity of hypoxic-ischemic encephalopathy and altered thyroid hormone and TSH levels, and between mortality and FT4 levels in the arterial blood of newborns between 18 and 24 h of life. METHODS: A case-control study was carried out. The case group comprised 17 term newborns (Apgar score < or = 3 and < or = 5 at the first and fifth minutes; umbilical cord blood pH < or = 7.15) who required bag and mask ventilation for at least one minute immediately after birth. The control group consisted of 17 normal, term newborns (Apgar score > or = 8 and > or = 9 at the first and fifth minutes; umbilical cord blood pH > or = 7.2). Cord blood and arterial blood samples were collected immediately after birth and 18 to 24 h after birth, respectively, and were used in the blood gas analysis and to determine serum concentrations of T4, T3, FT4, rT3 and TSH by radioimmunoassay. All newborns were followed-up until hospital discharge or death. RESULTS: Gestational age, birthweight, sex, size for gestational age, mode of delivery and skin color (white and non-white) were similar for both groups. No differences were found in mean levels of cord blood TSH, T4, T3 and FT4 between the groups. In the samples collected 18 to 24 h after birth, mean levels of TSH, T4, T3 and FT4 were significantly lower in the asphyxiated group than in the control group. Mean concentrations of arterial TSH, T4 and T3 between 18 and 24 h of life were lower than concentrations found in the cord blood analysis in asphyxiated newborns, but not in controls. In addition, asphyxiated newborns with moderate/severe hypoxic ischemic encephalopathy presented significantly lower mean levels of TSH, T4, T3 and FT4 than those of controls. None of the asphyxiated newborns with FT4 > or = 2.0 ng/dl died; 6 out of the 11 asphyxiated newborns with FT4 < 2.0 ng/dl died. CONCLUSIONS: Serum concentrations of TSH, T4, T3 and FT4 are lower in asphyxiated newborns than in normal newborns between 18 and 24 h of life; this suggests central hypothyroidism secondary to asphyxia. Asphyxiated newborns with moderate/severe hypoxic-ischemic encephalopathy present a greater involvement of the thyroid function and consequently a greater risk of death. PMID- 12725551 TI - In-hospital mortality of newborn infants born before 33 weeks of gestation depends on the initial level of neonatal care: the EPIPAGE study. AB - AIM: To determine the relation between the level of initial neonatal care and in hospital mortality of infants born before 33 wk of gestation in the era of surfactant therapy. METHODS: A 1 y prospective population-based survey was conducted in the north of France, as part of the EPIPAGE (Epidemiologie des Petits Ages Gestationnels) survey. Perinatal data were recorded for 585 very premature newborns transferred to a neonatal intensive care unit in 1997. The relation between the level of the neonatal unit that provided care for the first consecutive 48 h and in-hospital mortality was assessed by multivariate logistic regression, and adjusted for perinatal data and initial disease severity, estimated by the Clinical Risk Index for Babies (CRIB). RESULTS: The average gestational age (mean +/- SD) was 31.6 +/- 0.62 wk in level I, 30.7 +/- 0.21 in level II, 29.9 +/- 0.13 in non-teaching level III, and 29.0 +/- 0.15 in the level III teaching unit (p < 0.0001). The mean in-hospital mortality rate was 8.4% and did not differ by level of care (p(trend) = 0.17). After adjustment for perinatal data and CRIB, however, with the teaching unit as the reference, the risk of death was significantly higher in level I-II units [adjusted odds ratio (ORa) = 7.9, 95% confidence interval (95% CI) 2.2-29.1], but not in the non-teaching level III units (ORa = 0.8, 95% CI 0.3-2.1). CONCLUSION: In-hospital mortality in non-teaching level III units was similar to that in a teaching unit, but significantly higher in level I-level II units. Neonatal care of newborns delivered before 33 wk of gestation should initially occur in level III units. PMID- 12725552 TI - Relationship between prenatal infant feeding intention and initiation and duration of breastfeeding: a cohort study. AB - AIM: To report the relationship between maternal prenatal intention to breastfeed and the actual initiation and duration of breastfeeding. METHODS: Pregnant women resident within Avon, UK, expected to give birth between 1 April 1991 and 31 December 1992 were recruited in a longitudinal cohort study. Main outcome measures included maternal infant feeding intention (breastfeed, breast and bottle feed, bottle feed, or uncertain) at 32 wk of pregnancy: intention in the first week, intention for the rest of the first month and intention in months 2 to 4; initiation and duration of breastfeeding up to six months. RESULTS: Data were available on 10,548 women. Prenatal intention to breastfeed had an influence on both initiation and duration of breastfeeding. Of the women intending to bottle feed from birth, only 3.4% initiated breastfeeding compared with 96.6% of women planning to breastfeed for at least four months. At six months postpartum, the mean duration of breastfeeding for women intending to breastfeed for at least five months was 4.4 mo (95% CI 4.3, 4.4), compared with 2.5 mo (95% CI 2.4, 2.6) for women with a prenatal intention to breastfeed for only one month. Logistic regression, using intended duration as the only explanatory variable, correctly predicted 91.4% of breastfeeding initiation and 72.2% of infant feeding at six months. CONCLUSIONS: This large population-based study confirms the strength of the relationship between maternal prenatal intention to breastfeed and both breastfeeding initiation and duration. Maternal intention was a stronger predictor than the standard demographic factors combined. This should be taken into account in future research, and trials should be undertaken to establish whether interventions could alter maternal intention and thereby increase rates of breastfeeding initiation and duration. PMID- 12725553 TI - Monitoring breastfeeding rates in Italy: national surveys 1995 and 1999. AB - AIM: To assess and compare the rates of initiation and duration of breastfeeding in Italy in 1995 and 1999, and to examine the adherence to the ten steps to successful breastfeeding recommended by WHO. METHODS: Two cohorts of mothers who delivered healthy infants in November 1995 (n = 1601) or November 1999 (n = 2450) were interviewed by telephone within 4 wk of delivery and when their infant were 3, 6, 9 and 12 mo of age. Type of breastfeeding was classified according to the WHO definitions. Adherence to the WHO ten steps was evaluated as experienced by the mothers. RESULTS: Initiation and duration of breastfeeding increased during the 1995-1999 period (p < 0.0001). The rate of breastfeeding at birth, at discharge and when the infants were 3, 6, 9 and 12 mo of age was 85%, 83%, 42%, 19%, 10% and 4%, in 1995 and 91%, 89%, 66%, 47%, 25% and 12% in 1999. The rate of exclusive breastfeeding was higher in 1999 than 1995 at birth (39% vs 30%, p < 0.0001) and at discharge (77% vs 70%, p < 0.0001), but overall no longer duration was observed in 1999. At 4 and 6 mo of age the rate of exclusive breastfeeding was 25% and 8% in 1995 and 31% and 5% in 1999. The adherence to each WHO step was higher in 1999 than in 1995 (p < 0.05) but concomitant adherence was low (<3%). CONCLUSION: Although an increase in initiation and duration of breastfeeding occurred in Italy during the 1995-1999 period, the duration of breastfeeding and exclusive breastfeeding is currently unsatisfactory, as is adherence to the ten steps specified by WHO. Promotion of breastfeeding and education and improvement in adherence to the WHO recommendations are still needed in Italy. PMID- 12725554 TI - Gender differences in outcome after neonatal intensive care: speech and language skills are less influenced in boys than in girls at 6.5 years. AB - AIM: To study language development at age 6.5 y in 230 children who had required neonatal intensive care (NIC) and 71 full-term neonatally healthy control children, all born in 1986-1989, with a focus on comparison between genders. METHODS: Eight aspects of spontaneous speech, 3 fine motor functions, 10 linguistic areas, Peabody Picture Vocabulary Test, and digit recall (ITPA) were assessed. RESULTS: Achievements for gender and gestational age groups were analysed (group I, 23-31 wk; subgroup IA, 23-27 wk; IB, 28-31 wk; group II, 32-36 wk; group III, >36 wk), with children with congenital malformations as a separate group. As a group, at 6.5 y NIC girls had more developed spontaneous speech than NIC boys, and performed better than NIC boys in some linguistic areas. NIC girls of group I had lower results than control girls in spontaneous speech aspects such as speech motor function, interaction and motivation, and in many areas of linguistic skills. In contrast, NIC boys of group I had higher results than control boys in auditory memory, and only regarding information and speech motor function in spontaneous speech were their results lower. Analysis of results of matched pairs of group I NIC girls versus control girls and group I NIC boys versus control boys revealed even more marked differences between NIC girls and their matched controls than between NIC boys and their matched controls. CONCLUSION: Language development in extremely or very preterm NIC boys seems to be less influenced than that in NIC girls by the need for and administration of NIC. The neonatal factors associated with low scores in girls are different from those in boys. PMID- 12725555 TI - Strabismus in infants of opiate-dependent mothers. AB - AIM: At a follow-up clinic for infants of opiate-dependent mothers it was noted that more infants than expected developed strabismus. This study aimed to assess the prevalence of strabismus and the need for active strabismus surveillance in this population. METHODS: Consecutive infants of opiate-dependent mothers born over an 18 mo period were recalled for ophthalmological assessment by an ophthalmologist and orthoptist. Those unable to attend were surveyed by telephone using a questionnaire. RESULTS: 49 (69%) of the 71 eligible infants were recalled at a mean age of 21 mo (range 6-39); 29 had a full ophthalmological examination and the remaining 20 completed the questionnaire only. Seven (14%) of the 49 recalled infants had strabismus on examination; 4 needed glasses or patching. A further seven (14%) had a history of intermittent strabismus but declined formal examination. Another child had significant hypermetropia without strabismus. The mean age at which strabismus was observed was 8.3 mo (range birth to 19 mo). The presence of strabismus was not significantly influenced by conditions at birth, maternal drug doses, family history or need for or duration of abstinence treatment. CONCLUSION: The rate of strabismus in infants of opiate-dependent mothers was at least 10 times that in the general population. As attendance at follow-up is often poor, paediatricians should be aware of the association to encourage opportunistic assessment and ophthalmological surveillance of this population. PMID- 12725556 TI - Analysis of the mitochondrial genome in sudden infant death syndrome. AB - AIM: To investigate the mitochondrial genome and its association with sudden infant death syndrome (SIDS). METHODS: Twenty SIDS infants were screened for previously reported mitochondrial DNA mutations using direct sequencing. The whole mitochondrial genome was also sequenced for six of the infants. RESULTS: Three substitutions, A11467G, A12308G and G12372A, comprising a haplogroup were present in four infants diagnosed as pure SIDS. This haplogroup was also common in a control group. CONCLUSIONS: No specific mutation or polymorphism was found in association with SIDS. PMID- 12725557 TI - Congenital rickets due to maternal vitamin D deficiency in a sunny island of Greece. AB - A full-term male infant presented with clinical and biochemical findings consistent with the diagnosis of congenital rickets: weak muscle tone, craniotabes, episodes of tremor, hypocalcaemia, elevated serum alkaline phosphatase, secondary hyperparathyroidism, decreased 25-hydroxyvitamin D and normal 1,25-dihydroxyvitamin D serum levels. The mother's history and biochemical findings suggested nutritional vitamin D deficiency. Treatment with calcium and vitamin D resulted in the disappearance of clinical findings of rickets, normalization of the baby's biochemical profile and normal growth. It is surprising that this case occurred in an affluent setting, in the Mediterranean island of Crete, with an abundance of sunlight throughout the year. CONCLUSION: We report this case in order to emphasize the continuing occurrence of congenital rickets even in populations not considered at risk for hypovitaminosis D. A high index of suspicion is required for prompt diagnosis and treatment, thus preventing complications. PMID- 12725558 TI - Congenital tuberculous lymphadenitis in a preterm infant in Greece. AB - Congenital tuberculosis is a rare disease. The risk of tuberculosis in pregnancy has increased owing to recent changes in the epidemiology of the disease, which have led to an increased risk of congenital tuberculosis. We present a case report on a 6-d-old premature infant with tuberculous lymphadenitis. Smears of the lymphatic tissue contained acid-fast bacilli, and cultures were positive for Mycobacterium tuberculosis hominis. Polymerase chain reaction (PCR) assay of the suppurative material of the lymph node was positive for M. tuberculosis. Twenty days before onset of labour, the mother developed miliary tuberculosis and meningitis. CONCLUSION: The atypical clinical manifestations of congenital tuberculosis and the devastating consequences in the absence of early therapy signify the importance of early diagnosis and treatment during the neonatal period. PCR assay is a useful technique for prompt diagnosis in neonates with clinically suspected infection. PMID- 12725559 TI - Intra-rachidian disorders: two unusual cases of recurrent abdominal pain in children. AB - Recurrent abdominal pain (RAP) is generally thought to be of psychological origin but organic aetiologies are increasingly being identified. The cases of two children with vertebral disorders revealed by recurrent abdominal pain are reported. A 14-y-old girl presented with RAP associated with scoliosis, due to a T8-T9 intra-dural extra-medullary tumour. A 7-y-old girl who suffered from nocturnal RAP located in the right iliac fossa for 1 y had decreased muscular strength, pyramidal signs, and a 10 degree Lassegue sign in the right lower limb, revealing a vascular malformation extending from T12 to L2. CONCLUSION: Atypical pain and associated neurological signs or scoliosis must raise the possibility of intra-rachidian disorders. Magnetic resonance examination will then precisely state the location and nature of the pathological process, avoiding excessive delay in therapeutic management. PMID- 12725560 TI - Acute inflammatory demyelinating polyradiculoneuropathy associated with perforin deficient familial haemophagocytic lymphohistiocytosis. AB - This study reports the first paediatric case of acute inflammatory demyelinating polyradiculoneuropathy (AIDP) associated with a fatal haemophagocytic lymphohistiocytosis (HLH). The patient developed progressive weakness of the lower limbs in the context of a picture of infectious mononucleosis and Epstein Barr virus (EBV) infection. After an apparent improvement, a fulminant hepatic failure and pancytopenia ensued, leading to death. Molecular genetic studies documented a compound heterozygosity for two mutations in the perforin (PRF1) gene as the background defect for a familial haemophagocytic lymphohistiocytosis (FHL). CONCLUSION: In this patient EBV infection triggered both AIDP and FHL. The latter condition was due to PRF1 deficiency. Two novel mutations in the PRF1 gene were concomitantly present in the patient. The first caused an amino acid change, while the second introduced a stop codon in the sequence which resulted in a truncated protein. PMID- 12725561 TI - A randomized trial of two different doses of a SHR-5 Rhodiola rosea extract versus placebo and control of capacity for mental work. AB - A randomized, double-blind, placebo-controlled, parallel-group clinical study with an extra non-treatment group was performed to measure the effect of a single dose of standardized SHR-5 Rhodiola rosea extract on capacity for mental work against a background of fatigue and stress. An additional objective was to investigate a possible difference between two doses, one dose being chosen as the standard mean dose in accordance with well-established medicinal use as a psychostimulant/adaptogen, the other dose being 50% higher. Some physiological parameters, e.g. pulse rate, systolic and diastolic blood pressure, were also measured. The study was carried out on a highly uniform population comprising 161 cadets aged from 19 to 21 years. All groups were found to have very similar initial data, with no significant difference with regard to any parameter. The study showed a pronounced antifatigue effect reflected in an antifatigue index defined as a ratio called AFI. The verum groups had AFI mean values of 1.0385 and 1.0195, 2 and 3 capsules respectively, whilst the figure for the placebo group was 0.9046. This was statistically highly significant (p < 0.001) for both doses (verum groups), whilst no significant difference between the two dosage groups was observed. There was a possible trend in favour of the lower dose in the psychometric tests. No such trend was found in the physiological tests. PMID- 12725562 TI - Induction of NGF synthesis in astrocytes by onjisaponins of Polygala tenuifolia, constituents of kampo (Japanese herbal) medicine, Ninjin-yoei-to. AB - The effect of a kampo medicine, Ninjin-yoei-to (NYT; Ren-shen-yang-rong-tang in Chinese) on nerve growth factor (NGF) secretion from the cultured rat astrocytes was examined in vitro. When rat embryo astrocytes were cultured in the presence of NYT for 24 h, the amount of NGF in the medium was significantly increased in a dose dependent manner. Among 14 kinds of component herbs in NYT, the roots of Polygala tenuifolia and roots of Panax ginseng extracts increased NGF levels from the astrocytes. Saponin fraction from the roots of P. tenuifolia enhanced the production of NGF, however phenolic glycoside fraction showed no effect. Onjisaponins A, B, E, F and G as major saponins of the root of P. tenuifolia strongly increased the NGF level, whereas ginsenosides Rb1 and Rg1 did not affect the NGF level. Onjisaponin F also induced ChAT mRNA level in rat basal forebrain cells. These results indicate the possibility that NYT and/or onjisaponins in P. tenuifolia may have potential therapeutic effects for the treatment of Alzheimer disease patients. PMID- 12725563 TI - Cardiovascular, antihyperlipidemic and antioxidant effects of oleanolic and ursolic acids in experimental hypertension. AB - Cardiovascular (systolic and diastolic blood pressure, heart rate), antihyperlipidemic (tryglycerides, total cholesterol and lipoprotein fractions), antioxidant (glutathione peroxidase--GPx, and superoxide dismutase--SOD), diuretic/saluretic and hypoglycemic activity of 98% pure oleanolic (OA) and ursolic (UA) acid were studied in Dahl salt-sensitive (DSS), insulin resistant rat model of genetic hypertension. Both OA and UA displayed low toxicity, with LC50 0.10 and 0.95 mg/ml, respectively. Although both triterpenoids did not have direct hypotensive effect, after 6-week application in a daily dose 60 mg/kg b.w., i.p., they prevented the development of severe hypertension. The antihypertensive effect was attributed to their potent diuretic-natriuretic saluretic activity; direct cardiac effect (heart rate decrease by 34% and 32%, respectively); antihyperlipidemic (more than two times decrease of LDL and triglycerides); antioxidant (GPx increase by 12% and 10%, respectively; SOD increase by 12% and 22%, respectively), and hypoglycemic (blood glucose decrease by 20% and 50%, respectively) effects on the DSS rats. Except for the antihyperlipidemic effects, the other described above in vivo antihypertensive effects of OA and UA are reported for the first time and the underlying mechanisms are currently under investigation. PMID- 12725564 TI - Measurement of plasma procyanidin B-2 and procyanidin B-3 levels after oral administration in rat. AB - Using a high-performance liquid chromatographic method and mass spectrometry analysis, we successfully measured the absorption of orally administered procyanidin B-2 and procyanidin B-3 isolated from Cinnamonomi cortex (the bark of Cinnamomum cassia Blume) in the rat plasma. This method used a TSK- GEL ODS-80TS column, two solvents (A: 0.01% acetic acid; B: methanol with 0.01% acetic acid) in a linear gradient at a flow-rate of 1.0 ml/min, and fluorescence detection at excitation and emission wavelengths of 220 and 327 nm. PMID- 12725566 TI - Potato disc tumor induction assay: a multiple mode of drug action assay. AB - The study reported herein utilized the Agrobacterium tumefaciens-induced potato disc tumor assay. The objective was to verify the detection of antineoplastic activity in the potato disc tumor induction assay, regardless of the mode of antineoplastic drug action. Camptothecin, paclitaxel, podophyllin, vinblastine and vincristine were tested, each with a different mode of action. All drugs tested inhibited tumor induction. The order of activity was: camptothecin = paclitaxel = vinblastine < podophyllin = vincristine. No effect on the viability of the bacterium was detected. The A. tumefaciens-induced potato disc tumor assay was an effective indicator of antitumor activity regardless of the mechanism of drug action. Thus, this assay would be acceptable as a primary general screen for antineoplastic activity of various crude extracts, as well as for purified fractions, regardless of mode of inhibitory action on tumor formation. PMID- 12725565 TI - Effect of the crude extract of Vernonia polyanthes Less. on blood pressure and renal sodium excretion in unanesthetized rats. AB - This study evaluated the effect of oral crude Vernonia polyanthes Less. hydroalcoholic extract administration (CHE, 0.5 and 1.0 g/kg body wt., daily for 7 days) on arterial blood pressure and renal sodium excretion in conscious rats. CHE administration decreased arterial blood pressure dose-dependently followed by a significant rise in creatinine clearance and a fall in fractional post-proximal sodium excretion was compared to the control group. These results suggest that blood pressure decrease induced by the oral crude Vernonia hydroalcoholic extract may be blunted by reduction of the post-proximal renal sodium excretion. Thus, the present study shows that Vernonia extract is a potential vasodilatation agent in normotensive rats without any effects on renal tubule autoregulation mechanisms. PMID- 12725567 TI - Evaluation of the mutagenic, antimutagenic and antiproliferative potential of Croton lechleri (Muell. Arg.) latex. AB - Sangre de Drago is a red viscous latex extracted from Croton lechleri (Euphorbiaceae) cortex, renowned in South American popular medicine for its wound healing properties. The in vitro antiproliferative effects were determined on the human myelogenous leukemia K562 cells line (IC50 = 2.5 +/- 0.3 microg ml(-1)). The mutagenic and antimutagenic activity of C. lechleri sap was examined by means of the Ames/Salmonella test. No mutagenic activity was found on the Salmonella typhimurium strains T98 and T100, either with or without S9 activation. On the other hand, the sap showed an inhibitory effect against the mutagenic activity of the indirectly acting mutagen 2-Aminoanthracene in presence of S9 and a moderate protective activity against directly acting mutagens Sodium Azide and 2 Nitrofluorene. Therefore we suggest that C. lechleri sap interacts with the enzymes of the S9 mix, thereby inhibiting the transformation of 2-Aminoantracene into its active forms. PMID- 12725568 TI - Effect over time of in-vivo administration of the polysaccharide arabinogalactan on immune and hemopoietic cell lineages in murine spleen and bone marrow. AB - Current evidence indicates an immunostimulating role for complex carbohydrates, i.e., polysaccharides, from several plant sources. In the present work, we determined the specific in vivo effects, with time of administration, of one such compound, a neutral arabinogalactan from larch not only on immune (lymphoid) cells, but also on natural killer (NK) lymphoid cells, as well as a variety of other hemopoietic cells in both the bone marrow and spleen of healthy, young adult mice. The latter were injected daily (i.p.) with arabinogalactan (500 microg in 0.1 ml pH 7.2 phosphate buffered saline-PBS) for 7 or 14 days. Additional, aged (1 1/2-2 yr) mice were similarly injected for 14 days only. Control mice were given the PBS vehicle in all cases, following the above injection regimen. Animals from all groups were sampled 24 h after the final injection and the immune and hemopoietic cell populations in the bone marow and spleen were assessed quantitatively. The results indicated that immediately following either 7 or 14 days of arabinogalactan administration to young, adult mice, lymphoid cells in the bone marrow were significantly decreased (p < 0.004; p < 0.001, respectively) relative to controls but remained unchanged at both time intervals in the spleen. NK cells, after 7 days of arabinogalactan exposure, were also decreased significantly in the bone marrow (p < 0.02), but unchanged in the spleen. After 14 days' exposure to the polysaccharide, NK cells in the bone marrow had returned to normal (control) levels, but were increased in the spleen (p < 0.004) to levels greater than 2-fold that of control. Among other hemopoietic cell lineages, none was influenced in the bone marrow or spleen by one-week administration of arabinogalactan; however, after two-week exposure, precursor myeloid cells and their mature (functional) progeny (granulocytes), were significantly reduced in the spleen (p < 0.043; p < 0.006, respectively), as were splenic monocytes (p < 0.001). These lineages in the bone marrow, however, remained steadfastly unaltered even after 14 days of continuous exposure to the agent. Of the vast cascade of cytokines induced in the presence of this polysaccharide, it appears that immunopoiesis- and hemopoiesis-inhibiting ones are most prevalent during at least the first two weeks of daily exposure. PMID- 12725569 TI - Immunomodulatory activity of Mollugo verticillata L. AB - This article describes the evaluation of immunomodulatory activity of Mollugo verticillata L. (Molluginaceae), a weed plant common in warm and/or wet regions of the American continent. Nitric oxide (NO) release was evaluated in mice peritoneal cell cultures treated in vivo using the ethanolic extract of M. verticillata with and without BCG. The plant extract showed immunostimulatory activity when peritoneal cells were stimulated in vitro with BCG antigen only. However, mice peritoneal cells treated with M. verticillata plus BCG showed a drastic reduction in NO production when they received the additional stimulus in vitro with BCG. Ethanolic extracts of M. verticillata could directly increase NO release by peritoneal cells, but suppress the immune response of these cells when treated with BCG antigen and Mycobacterium tuberculosis whole antigen (TB). Preliminary phytochemical tests allowed the detection of quercetin and triterpenoid glycosides in the ethanolic extract of M. verticillata, and those compounds are probably responsible for the effect of this plant material on the immune system. PMID- 12725570 TI - Effect of beta-carotene on the inhibition of lung metastasis in mice. AB - Effect of beta-carotene on the inhibition of lung metastasis induced by B16F-10 melanoma cells was studied in C57BL/6 mice. Simultaneous administration of the compound after tumor induction produced a significant reduction (71.36%) in tumor nodule formation. Increased lung collagen hydroxyproline (22.37 microg/mg protein) in the metastasized lungs of control animals compared to the normal animals (0.95 microg/mg protein) was significantly reduced (4.19 microg/mg protein) in the beta-carotene treated animals. High amount of uronic acid (355.83 microg/100mg tissue ) in the metastasized control animals was significantly reduced (87.87 microg/100 mg tissue) in the animals treated with beta-carotene. Lung hexosamine content also was inhibited significantly in the beta-carotene treated animals (1.58 mg/100 mg lyophilized tissue) compared to the untreated control animals (4.2 mg/100 mg lyophilized tissue). The elevated levels of serum sialic acid and serum gamma glutamyl transpeptidase activity in the untreated control animals was significantly reduced in the animals treated with beta carotene. Beta carotene treated animals were survived up to 69 days. Histopathology of the lung tissue also correlated with the above parameters and life span of the drug treated animals. Our results reveal the antimetastatic activity of beta-carotene which are abundantly present in green plants, vegetables and fruits. PMID- 12725571 TI - Hepatoprotective and free radical scavenging effects of Nelumbo nucifera. AB - Ethanol extracts from Nelumbo nucifera (ENN) seeds were studied for possible antioxidative and hepatoprotective effects. Antioxidative effects were measured spectrophotometrically by reduction of 2,2'-Diphenyl-1-picrylhydrazyl (DPPH) radicals. Hepatoprotective effects were tested using carbon tetrachloride (CCl4) and aflatoxin B1 (AFB1)-induced hepatocyte toxicity models. ENN showed potent free radical scavenging effects with a median inhibition concentration of 6.49 microg/ml. Treatment of hepatocytes with ENN inhibited both the production of serum enzymes and cytotoxicity by CCl4. The genotoxic and cytotoxic effects of AFB1 were also inhibited by ENN in dose-dependent manners. These hepatoprotective effects of ENN against CCl4 and AFB1 might result from its potent antioxidative properties. PMID- 12725572 TI - Free radical-scavenging activity of Taiwanese native plants. AB - The 70% aqueous acetone extracts of ten Taiwanese native plants were evaluated by various antioxidant assays, including 1, 1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl (.OH) radicals, and reducing power assay. In the present study, extracts of Acer buerferianum var. formosanum, Cleyera japonica var. morii, Cyclobalanopsis stenophylla var. stenophylloides, and Machilus zuihoensis exhibited stronger activity against DPPH radicals, and their IC50 values ranged from 5.4 to 8.3 microg/ml. The ten selected extracts effectively inhibited the formation of .OH generated in the Fenton reaction system. Among the extracts whose reducing power activities were determined, A. buerferianum var. formosanum, C. japonica var. morii, C. stenophylla var. stenophylloides, Eriobotrya deflex, and M. zuihoensis showed high activity. The results indicate the 70% aqueous acetone extracts of A. buerferianum var. formosanum, C. japonica var. morii, C. stenophylla var. stenophylloides, and M. zuihoensis with great potency in these assay systems and may be candidates for the development of natural antioxidants. PMID- 12725573 TI - Anti-inflammatory and antioxidant effects of Croton celtidifolius bark. AB - Croton celtidifolius Baill commonly known as "sangue-de-adave" is a tree found in the Atlantic Forest of south of Brazil, mainly in Santa Catarina. The bark and leaf infusions of this medicinal plant have been popularly used for the treatment of inflammatory diseases. In this study we evaluated the anti-inflammatory and antioxidant properties of crude extract (CE), aqueous fraction (AqF), ethyl acetate fraction (EAF), butanolic fraction (BuF) and catechin, gallocatechin and sub-fractions, 19SF, 35SF and 63SF that contained a mixture of proanthocyanidins and were derived from the EAF fraction. The CE, AqF, EAF, BuF, catechin and sub fractions 35SF and 63SF reduced paw edema induced by carrageenan. The CE, fractions, sub-fractions and isolated compounds showed antioxidant properties in vitro, all were able to scavenge superoxide anions at a concentration of 100 microg ml(-1). The EAF, catechin and gallocatechin were most effective in the deoxyribose assay, IC50 0.69 (0.44-1.06), 0.20 (0.11-0.39), 0.55 (0.28-1.08) microg x ml(-1) respectively. The CE and other fractions and sub-fractions inhibited deoxyribose degradation up to 1 microg x ml(-1). In the hydrophobic system only AqF did not show lipid peroxidation inhibition. The CE, other fractions, sub-fractions and isolated compounds inhibited lipidid peroxidation only at a concentration of 100 microg x ml(-1). In summary, this study demonstrates that Croton celtidifolius bark has significant anti-inflammatory and antioxidant activity. PMID- 12725574 TI - Evaluation of anti-inflammatory activity of Vernonia cinerea Less. extract in rats. AB - The methanol extract of the whole plant of Vernonia cinerea Less. was evaluated for its anti-inflammatory activity in acute (carrageenin, histamine and serotonin induced rat paw edema) and a chronic model (cotton pouch induced granuloma). The methanol extract (250 and 500 mg/kg(-1) p.o.) exhibited significant activity (p < 0.001) against all phlogistic agents used in a dose dependant manner. In the chronic model (cotton pouch granuloma method) the methanol extract exhibited significant anti-inflammatory activity. All these effects were compared with standard drug phenylbutazone (100 mg/kg(-1) p.o.). PMID- 12725575 TI - Analgesic and antiinflammatory effects of chalcones isolated from Myracrodruon urundeuva allemao. AB - The present work showed analgesic and antiinflammatory activities from a fraction containing three dimeric chalcones (chalcone enriched fraction - CEF), isolated from the stem-bark ethyl acetate extract of Myracrodruon urundeuva Allemao (Anacardiaceae). M. urundeuva is a popular medicinal plant used widely in Northeast Brazil, mainly as a topical female genital tract antiinflammatory. We observed that the CEF (5 and 10 mg/kg body wt., i.p. or p.o.) inhibited acetic acid-induced abdominal contractions in mice. In the formalin test, the CEF (5 and 10 mg/kg body wt.) was more effective intraperitoneally and inhibited predominantly the second phase of response. Naloxone reversed this effect, indicating an involvement of the opioid system. The CEF (10 and 20 mg/kg body wt.) also increased the reaction time to thermal stimuli in the hot-plate test in mice, after i.p. but not after p.o. administration. In the carrageenan-induced paw edema test in mice, the CEF (20 and 40 mg/kg body wt.) decreased paw volume significantly, after i.p. administration 2-4 hours after carrageenan injection. The CEF (40 mg/kg body wt.) was also active orally during the same period of time. The present work is the first report on peripheral and central analgesic effects and antiinflammatory activity of natural dimeric chalcones. PMID- 12725576 TI - Dose dependent hypoglycemic effect of aqueous extract of Enicostemma littorale blume in alloxan induced diabetic rats. AB - Previous studies in our lab had confirmed the blood glucose lowering effect of E. littorale Blume in alloxan induced diabetic rats with no change in normoglycemic control rats. Present paper deals with dose dependent (0.5, 1.0, 1.5, 2.5, 3.5 g dry plant equivalent extract/100 g body wt., p.o.) blood glucose lowering effect of aqueous extract of E. littorale Blume in alloxan induced diabetic rats. The effective dose was found to be 1.5 g dry plant equivalent extract/100 g body wt.. The above dose caused significant decrease in glycosylated haemoglobin, liver glucose-6-phosphatase activity and significant increase in serum insulin levels of the diabetic rats. No significant changes were observed in the toxicity parameters of extract treated diabetic rats as compared to diabetic control rats. The above results suggest that E. littorale is a potent antidiabetic agent without any toxic effect at this particular dose (1.5 g dry plant equivalent extract/100 g body wt.). PMID- 12725577 TI - Determination of huperzine A in formulated products by reversed-phase-liquid chromatography using diode array and electrospray ionization mass spectrometric detection. AB - A precise and selective high-performance liquid chromatographic (HPLC) method with diode-array detection for quantifying huperzine A in formulated products was developed and validated. A liquid chromatographic-mass spectrometric (LC/MS) procedure was devised to confirm the HPLC method. Huperzine A was dissolved in 1,2-dichloroethane, chromatographed on a YMCBasic C18 column, and detected at 308 nm. A gradient mobile phase of 10 mM ammonium acetate (pH = 3.5)--methanol was used. Identification was based on retention time, UV spectra and mass spectra by comparison with a commercial standard. The UV peak areas were used for quantitation of huperzine A content. The correlation coefficient (R2) of the calibration curve was 1 over the range 0.8-11.6 microg/ml. Overall recovery of huperzine A was 103.9% +/- 1.8 (mean +/- SD). Relative standard deviations for intra- and interday precision were < 2%. PMID- 12725578 TI - Treatment with Hypericum perforatum L. does not trigger decreased resistance in Staphylococcus aureus against antibiotics and hyperforin. AB - Most scientific investigations of Hypericum perforatum L. (Saint John's wort) concentrated on its antidepressant activity. Only recently, its antibacterial activity against multiresistant Staphylococcus aureus led to speculations regarding the use of hyperforin, an antibacterial principle of hypericum, as an antibiotic. In the present investigation, we show that Staphylococcus aureus is able to acquire a resistance against hyperforin which did not lead to a cross resistance against clinically used antibiotics. Resistance development does not take place, however, at concentrations as low as they are found in human blood plasma during antidepressant treatment with 900 mg Hypericum extract/day. PMID- 12725579 TI - Biological screening of Annonaceous Brazilian Medicinal Plants using Artemia salina (brine shrimp test). AB - Eighteen different extracts from five Annona species collected in Minas Gerais state, Brazil, were submitted to the brine shrimp lethality test in order to detect potential sources of novel cytotoxic, antitumor, pesticidal and anti Trypanosoma cruzi compounds. All of the Annonaceous species tested showed good larvicidal activity as compared to a reference compounds and literature data. PMID- 12725580 TI - Efficacy of dry extract of ivy leaves in children with bronchial asthma--a review of randomized controlled trials. AB - OBJECTIVES: To investigate if extracts from dried ivy leaves (Hedera helix L.) are effective in the treatment of chronic airway obstruction in children suffering from bronchial asthma. DESIGN: Systematic review of trials documented in the literature with re-analysis of original data. TRIALS: 5 randomized controlled trials investigating the efficacy of ivy leaf extract preparations in chronic bronchitis, 3 of which were conducted in children and met our selection criteria. One compared ivy leaf extract cough drops to placebo, one compared suppositories to drops and one tested syrup against drops. MAIN OUTCOME MEASURES: Body-plethysmographic and spirometric measures. RESULTS: Drops were significantly superior to placebo in reducing airway resistance (primary outcome measure; p = 0.04 two-sided) and descriptively superior in all other 'objective' measures. For syrup and suppositories, at least 54%, resp. 35% of the effect against placebo were preserved. CONCLUSIONS: The trials included in this review indicate that ivy leaf extract preparations have effects with respect to an improvement of respiratory functions of children with chronic bronchial asthma, but more far reaching conclusions can hardly be drawn because of a meagre database, including the fact that only one primary trial included a placebo control. Further research, particularly into the long-term efficacy of the herbal extract, is needed. PMID- 12725582 TI - A review on potentiality of medicinal plants as the source of new contraceptive principles. AB - Recently, a review has already been made on the synthetic contraceptive agents whereas this review embraces the natural contraceptives upto year 2001 with 355 references. It also includes the isolation of their active principles, methods of analysis of active ingredients through TLC, HPLC, their side effects and pharmacological action. PMID- 12725581 TI - Natural product inhibitors of ovarian neoplasia. AB - The present work constitutes a review of the literature on natural products with potential antitumor activity against ovarian neoplasias. The review refers to five plant extracts and sixty-nine compounds isolated from higher plants and microorganisms, which are classified in appropriate chemical groups and model tested, and cites their activity. Some aspects of recent research with natural products directed to ward producing drugs which are inhibitors of ovarian neoplasia are discussed. PMID- 12725583 TI - Kava-kava: a lesson for the phytomedicine community. PMID- 12725584 TI - Dysmorphology in literature: describing is not condoning. PMID- 12725585 TI - Congenital malformations and genetic diseases in comic books. AB - Medical syndromes have often been represented in fine arts, but rarely have clinical diagnoses been discussed in comic book characters. Since their first appearance in Europe in the middle of the 19th century and in America in 1895, comic books have been considered as "the 9th art". In many comic books, the appearance and/or the behavior of central or support characters are suggestive of already well-defined medical disorders. The representation of five particular groups or clinical features: mental retardation, abnormal stature, abnormal hair, obesity, and cranial malformations is discussed from mostly European comic series. Whether comic authors intended to describe specific clinical entities while drawing their characters or whether such situations appeared by mere luck, is open to debate. In many series from the first half of the 20th century characters with remarkable clinical features were also painted as psycho-social deviants. Such stereotypes are found much less frequently nowadays. Writers of comic books, realizing the major impact of their work especially in adolescent age groups, have increasingly been using their series to actually promote issues of equity and well being for physically or mentally impaired people. PMID- 12725587 TI - A variant example of familial Floating-Harbor syndrome? AB - The Floating-Harbor syndrome (FHS) is clinically characterized by short stature, retarded speech development, delayed bone age, typical facies, bulbous nose, wide columella, thin lips. Four cases with celiac disease have been described previously. In two other cases, autosomal dominant inheritance has been suggested. We describe a boy aged 2 years 11 months with clinical features of FHS and celiac disease. His mother also presents minor phenotypical characteristics, suggesting that the present observation corresponds to a variant example of familial FHS. PMID- 12725586 TI - Pre-academic and early academic achievement in children with velocardiofacial syndrome (del22q11.2) of borderline or normal intelligence. AB - School-aged children with del22q11.2 tend to show a typical learning and neuropsychological profile, which is characterised by a VIQ-PIQ discrepancy (in favour of the VIQ) and significantly better scores for reading (decoding) and spelling compared to mathematics. To the best of our knowledge, there exists no systematic research on the pre-academic and early academic skills that might underpin these learning difficulties. The purpose of the current study was to investigate more systematically these pre-academic and early academic skills in borderline to normal intelligent (FSIQ > 70) children with del22q11.2 in the last year of kindergarten and first grade of primary school in Flanders. In the kindergarten group, meta-linguistic awareness and counting skills were examined. In the group of first graders, children were tested on reading, spelling and mathematics. Thirteen children (mean age: 6 years 4 months (SD = 0.84); 9 boys, 4 girls) participated in this study. In the present study, there were no differences in intelligence and academic outcomes between boys and girls, and no differences in IQ and academic achievement between children with cardiac defects or severe velopharyngeal insufficiency (VPI) and children without these deficits. With regard to pre-academic achievement in general, a characteristic profile with clearly better results for meta-linguistic awareness in comparison to counting skills was found, but this difference is not statistically significant. Concerning early academic achievement, children with del22q11.2, as a group, perform (despite their somewhat lower general intelligence) on average compared with their age-related peers. However, at an individual level--especially within the domain of counting skills and mathematics--there is a wide variability, with some children showing remarkable learning difficulties already at an early age. PMID- 12725588 TI - Triphalangeal thumb and psychomotor retardation: a new association? AB - The triphalangeal thumb (TPT) is a rare malformation in which the thumb is presented as a long digit of three phalanges. We describe two brothers showing TPT and psychomotor retardation, especially in language. Difficulties in language development were also observed in children with TPT in another study. The coexistence of TPT and psychomotor retardation in those patients and in the two patients described here suggests that TPT and psychomotor retardation could be part of a syndromic association not described previously. PMID- 12725589 TI - Clinical, cytogenetical and molecular analyses of Angelman syndrome. AB - A total of 95 patients suspected with the clinical diagnosis of AS were evaluated and 37 cases (39%) were confirmed by cytogenetic or molecular studies as affected by Angelman syndrome. The clinical analysis was performed according to a specific clinical protocol for the diagnosis of AS. Cytogenetical analysis was used to detect chromosome rearrangements by determining the karyotype in lymphocytes by GTG banding and revealed an abnormal karyotype in two cases (5.4%), both of them presenting a new pericentromeric inversion in chromosome 15. Molecular analyses included determination of DNA methylation within the 15q11-13 region by Southern blotting and microsattelite analysis within the 15q11-13 region by PCR and the UBE3A gene was also studied by mutational screening. In 16 cases (43.2%) a de novo deletion was detected in the maternal chromosome 15:3 cases (8.1%) presented imprinting defect at the 15q11-13 region; one case is due to a paternal uniparental dissomy (2.7%) and another two cases showed a inherited mutation at the UBE3A gene (5.4%). Thirteen cases (35.1%) showed no deletion, no UPD, no imprinting defect, no UBE3A mutation and the diagnosis of AS could be ruled out in 58 patients. The objective of the present work was to describe the clinical and laboratory protocols employed at our laboratory in order to establish the AS study. We conclude that the protocols employed here were efficient for the diagnosis of AS, a frequently underdiagnosed pathology. PMID- 12725590 TI - X-linked hydrocephalus: another two families with an L1 mutation. AB - X-linked hydrocephalus is a variable condition caused by mutations in the gene encoding for L1CAM. This gene is located at Xq28. Clinically the spectrum ranges from males with lethal congenital hydrocephalus to mild/moderate mental retardation and spastic paraplegia. Few carrier females show minimal signs of the syndrome. Although most cases are familial, de novo situations have been reported. We report two new families with the syndrome and a L1 mutation. Family 1 has two patients and family 2 a single patient. Clinical diagnosis in all three affected boys was beyond doubt. Prenatal testing through chorionic villus biopsy is possible only with a demonstrated L1 mutation. In lethal sporadic cases neuropathology is very important in order to evaluate for features of the syndrome. We stress the importance of further clinical reports including data on neuropathology and DNA analysis in order to further understand the mechanisms involved in this disorder. PMID- 12725591 TI - A phenotypically normal liveborn male after prenatal diagnosis of trisomy 20 mosaicism. AB - We report on a case of prenatally diagnosed true trisomy 20 mosaicism in amniocytes. Cytogenetic analysis was performed postnatally on lymphocytes and extra-embryonic tissues. For analysing uroepithelial cells we established a new cell nuclei preparation protocol for FISH (Fluorescence In Situ Hybridization). Trisomy 20 cells could not be confirmed after birth. The origin or trisomy 20 cells in amniotic fluid remains unclear. The phenotypically normal male baby is developing normal. PMID- 12725592 TI - Where to look for the genes related to diaphragmatic hernia? AB - Analysis of approximately 150 published observations of diaphragmatic hernia (DH) in persons with structural autosomal imbalance showed several segments where DH related genes may be found. Occurrence of DH in several patients with deletions 15q26, 8p23, 8q22, 4p16, 1q42, and 3q22 allows to propose that these segments harbor the genes which, when deleted (or truncated) may be responsible for DH. Segments 22q11, 4q28.3q32, 1q25q31.2 and 2p23p25 are good candidates for the location of genes which cause DH in trisomic condition. The genetic mechanisms of DH in tetrasomy 12p are not clear, although more than 50 cases of DH have been reported in this syndrome. Frequent coexistence of congenital heart defects and DH in some syndromes (and rarity of this association in some others) may suggest the different pathways of the DH's origin. PMID- 12725593 TI - Asymmetrical Larsen syndrome in a young girl: a second example of somatic mosaicism in this syndrome. AB - We describe another observation of asymmetrical Larsen syndrome. The unilateral manifestation of the typical skeletal defects indicates that this condition might be due to unilateral somatic cell-line mosaicism. PMID- 12725594 TI - Isolated hypertrophic pyloric stenosis and perinatal factors. AB - Infantile hypertrophic pyloric stenosis (IHPS) is a common condition requiring surgical intervention during the first weeks of life. Up to now the exact etiology of IHPS remains unclear and it is probable that several predisposing risk factors would be associated with the condition. Prompted by the observation that some perinatal factors may be involved in IHPS etiology, we evaluated 171 isolated cases referred to the Sicilian Registry of Congenital Anomalies. Our results show that some perinatal factors like sex ratio imbalance and parity are associated with IHPS, but further investigation is needed to clarify the relationship between genes and other factors involved in IHPS etiology. Therefore the presence of these perinatal factors may be accurately evaluated in genetic counseling to provide a perspective of recurrence prevention. PMID- 12725595 TI - Prenatal ultrasonographic diagnosis of a recurrent case of Johanson-Blizzard syndrome. AB - We report on ultrasonographic prenatal diagnosis of a recurrent case of Johanson Blizzard syndrome. The pregnancy was terminated at 21 weeks'. This observation highlights the great variability of the expression of this syndrome, including in the same family, and the necessity of collaborating with an experienced geneticist in all antenatal diagnosis of any complex of abnormalities. PMID- 12725596 TI - Goeminne syndrome (OMIM 314300): another male patient 30 years later. PMID- 12725597 TI - Asphyxiating thoracic dystrophy and syndactyly of the toes in a newborn: coincidence or a new association? PMID- 12725598 TI - A case of Marin-Amat syndrome. PMID- 12725599 TI - Recurrent hepatitis C in transplanted patients: more questions than answers. PMID- 12725600 TI - Sheep rearing and Helicobacter pylori infection--an epidemiological model of anthropozoonosis. PMID- 12725602 TI - Helicobacter pylori infection in subjects with acute ischaemic stroke. AB - AIMS: To determine whether infection with Helicobacter pylori is a significant risk factor for stroke. SUBJECTS: A total 467 in-patients with clinical evidence of acute ischaemic stroke and 388 healthy controls with no evidence of cerebrovascular disease. METHODS: This was a case control study. The prevalence of Helicobacter pylori was measured by enzyme-linked immunosorbent assay in stroke patients and controls. A positive titre was defined as >15 U/ml and relationship with circulating plasma fibrinogen and social depravation was expressed using the Townsend Index. RESULTS: There were significantly more Helicobacter pylori positive individuals (274/398 (69%)) in the cases compared to the controls (206/352 (58.5%)). Fibrinogen levels were also significantly higher in Helicobacter pylori positive (mean 4.14, standard deviation 1.33) than negative individuals (mean 3.78, standard deviation 1.28). The association between Helicobacter pylori and stroke was lost in a logistic model controlling for socio-economic status. Furthermore, fibrinogen levels were not associated with Helicobacter pylori status in a linear regression model controlling for socio-economic status. CONCLUSIONS: Infection with Helicobacter pylori is associated with an increased risk of stroke and increased fibrinogen levels but these findings can be attributed to a confounding effect of socio-economic status. PMID- 12725601 TI - Prevalence of Helicobacter pylori infection in Polish shepherds and their families. AB - BACKGROUND: Helicobacter pylori infection might, in some instances, be considered as zoonosis. AIM: The aim of this study was to assess the H. pylori prevalence in Polish shepherds and in their families as compared to controls. Patients and methods. A total of 42 shepherds from Polish Tatra Mountains with regular contact with sheep, 28 members of their families with incidental contacts and 61 age- and gender-matched farmer controls without such contacts were involved in this study. H. pylori status was determined by 13C-urea breath test. Serology was used to measure anti-H. pylori and anti-CagA IgG. Plasma gastrin, interleukin-8 and tumor necrosis factor-alpha were also determined. RESULTS: The H. pylori prevalence reached 97.6% in shepherds, 86% in their family members, but significantly less, 65.1%, in controls without contact with sheep. Anti-H. pylori IgG, anti-CagA in contact groups were significantly higher than in controls. Also, plasma gastrin, interleukin-8 and tumor necrosis factor-alpha had significantly higher values as compared to controls. CONCLUSIONS: Shepherds showed almost 100% H. pylori prevalence and higher incidence of CagA seropositivity, plasma gastrin and pro inflammatory cytokine levels. Considering 100% positive 13C-urea breath test in sheep, it may be reasonable to suggest that H. pylori infection in shepherds and their family members originates from sheep and H. pylori infection might, therefore, be considered as zoonosis. PMID- 12725603 TI - Triple therapy for 7 days vs. triple therapy for 7 days plus omeprazole for 21 days in treatment of active duodenal ulcer with Helicobacter pylori infection. A double blind placebo controlled trial. AB - BACKGROUND: The current treatment for active duodenal ulcer implies a 4 weeks course with anti-secretory drugs and two antibiotics for 7 to 10 days in the case of Helicobacter pylori infection. AIM: To establish whether triple therapy with omeprazole given for 7 days with two antibiotics eradicates H. pylori, heals and prevents ulcer recurrence. PATIENTS AND METHODS: A total of 103 patients with active duodenal ulcer and H. pylori were randomly divided into: a group of 50 patients treated with omeprazole, clarithromycin and tinidazole for 7 days followed by omeprazole for 21 days, and a group of 53 patients who received the same treatment as the previous group, followed by placebo for 21 days. Endoscopy with quick urease test, histology and culture was performed at entry and after 4 and 16 weeks. RESULTS: Ulcer healing rate after 16 weeks' treatment was 95% in the former and 96% in the latter group (ns). Eradication after 16 weeks was 84% in the former and 83% in the latter group (ns). At 56 weeks, all patients examined were ulcer free and without H. pylori. CONCLUSION: Omeprazole, clarithromycin and tinidazole for 7 days heals active duodenal ulcer and eradicates H. pylori infection in most patients. Treatment extension with omeprazole, for 3 weeks, after triple therapy does not modify healing and eradication rates. PMID- 12725604 TI - Cytology and image cytometry after colonic lavage: a complementary diagnostic tool in patients with ulcerative colitis. AB - BACKGROUND: Patients with extensive, long-standing ulcerative colitis have increased risk of colorectal cancer. AIMS: To improve the detection of high-risk patients, using a combination of colonic cytology, histology, and DNA image cytometry after segmental colonic lavage. PATIENTS: A series of 16 patients (8 high-risk patients) with ulcerative colitis were investigated. METHODS: After segmental lavage step, biopsies were obtained. Gradient centrifugation of the colonic fluid was performed for isolation and purification of epithelial cells. The smears and biopsy specimens obtained were stained for routine interpretation and for DNA image cytometry. RESULTS: Segmental lavage could be performed in all patients. Specimens from two high-risk patients showed low grade dysplasia and atypia by means of histology and cytology, respectively. In one patient, without increased colorectal cancer risk, atypia was detected. Three patients in the high risk group, two of those diagnosed as positive for dysplasia and atypia, showed aneuploidy histologically and cytologically. DNA aneuploidy, in cytological material, was found exclusively in three low-risk patients, one of those had atypia cytologically. CONCLUSIONS: Isolation and purification of epithelial cells after segmental colonic lavage using density gradient centrifugation can be performed as part of routine endoscopy. It provides information about atypical cells and DNA aneuploidy as additional markers of malignant transformation. The combination of cytologic examination and DNA image cytometry might improve the detection of high-risk ulcerative colitis patients. PMID- 12725606 TI - Tuberculous peritonitis in an endemic area. AB - BACKGROUND: Tuberculous peritonitis is a fatal disease if not diagnosed in time. AIMS: To identify the clinical, laboratory, and diagnostic features of tuberculous peritonitis in Iranian patients. PATIENTS: Included in the study were all cases of tuberculous peritonitis with a definite diagnosis confirmed by pathology in four referral University Hospitals in Tehran between 1989 and 1999. METHODS: All clinical, laboratory, and radiological findings as well as invasive procedures were reviewed. RESULTS: A total of 50 patients (30 female, 20 male), mean age 33.5 years were studied. Main presenting symptoms included abdominal pain (84%), weight loss (72%) and fever (50%). In 24% of patients a positive tuberculin test was found. Erythrocyte sedimentation rate >50 mm/h was detected in 60% of patients and 4.4% had an Erythrocyte sedimentation rate >100 mm/h. Laparoscopy or laparotomy showed peritoneal seeding in 74% of patients. CONCLUSIONS: Exudative ascites should give rise to clinical suspicion of tuberculous peritonitis in endemic areas or in immigrants from endemic areas. Laparoscopy is the most sensitive and specific diagnostic method. PMID- 12725605 TI - Val34Leu factor XIII polymorphism in Italian patients with inflammatory bowel disease. AB - BACKGROUND: Coagulation Factor XIII is implicated in fibrin stabilization and wound healing. Plasma levels of Factor XIII are reduced in inflammatory bowel disease patients; recently, a valine 34 to leucine polymorphism of the Factor XIII-A subunit gene with a defined protective effect against thrombosis and as yet undetermined effect on wound healing has been described. AIM: To evaluate Val34Leu Factor XIII polymorphism distribution and to find possible correlations with clinical features in Italian inflammatory bowel disease patients. STUDY POPULATION: A total of 152 inflammatory bowel disease patients, 90 with ulcerative colitis and 62 with Crohn's disease and 130 healthy volunteers were studied. METHODS: Val34Leu polymorphism was detected by RFLP with BsaH I. Statistical analysis was performed by means of Fisher exact test. RESULTS: In inflammatory bowel disease, 57.2% of patients showed the wild type status, 37.5% were heterozygous and 5.3% were homozygous for the 34Leu allele; the frequency of the mutated allele was 24.0%. In controls, 66.1% of subjects showed the wild type status, 28.5% were heterozygous and 5.4% were homozygous for the 34Leu allele; the frequency of the mutated allele was 19.7%. There was no difference in genotype distribution and prevalence of the mutated allele between inflammatory bowel disease patients and controls. CONCLUSIONS: The present data do not show any differences in Val34Leu Factor XIII polymorphism distribution between inflammatory bowel disease patients and controls. The prothrombotic state described in inflammatory bowel disease patients does not depend on an altered distribution of Val34Leu Factor XIII polymorphism. PMID- 12725607 TI - In patients with dermatitis herpetiformis distribution of transglutaminase in cutaneous tissue does not differ from controls. AB - BACKGROUND: Dermatitis herpetiformis may be regarded as the cutaneous counterpart of coeliac disease. These conditions are related to the ingestion of gluten and both are characterised by circulating antibodies against tissue transglutaminase. AIMS: To study the distribution of tissue transglutaminase in the skin of dermatitis herpetiformis patients and controls, and to investigate whether the dermal IgA deposits, diagnostic for dermatitis herpetiformis, are related to tissue transglutaminase expression in the skin. METHODS: A series of 11 patients with dermatitis herpetiformis had a 4 mm punch biopsy taken from the uninvolved perilesional skin. A group of 16 controls, undergoing surgical removal of benign nevi, gave perilesional skin. Biopsies were covered with OCT and frozen at -80 degrees C. After washing, skin biopsy sections were incubated with an IgG anti tissue transglutaminase mouse monoclonal antibody. After washing, sections were incubated with anti-mouse IgG. RESULTS: The anti-tissue transglutaminase monoclonal antibody specifically recognised the basal epidermal cells. This staining was no different between patients and controls. CONCLUSION: Our results suggest that tissue transglutaminase can be recognised in the basal epidermal layer both of patients with dermatitis herpetiformis and controls. Since this distribution does not correspond to the distribution of dermal IgA deposits, it is concluded that dermatitis herpetiformis dermal IgA deposits are not due to antibodies directed against cutaneous tissue transglutaminase. PMID- 12725608 TI - Reduced quality of life of patients with hepatocellular carcinoma. AB - BACKGROUND: It has been shown that health-related quality of life is reduced in patients with cirrhosis and with chronic hepatitis in relation to antiviral therapy. No data are available on patients with hepatocellular carcinoma. AIM: To assess health-related quality of life in cirrhotic patients with hepatocellular carcinoma. PATIENTS AND METHODS: Health-related quality of life was assessed in 101 hepatocellular carcinoma patients by means of Short Form-36 and Nottingham Health Profile questionnaires. Final scores of domains for individual patients were compared to age-adjusted normative Italian values, using Z-score and with values obtained in 202 matched patients with cirrhosis, without hepatocellular carcinoma. RESULTS: All Short Form-36 domains and 4 out of 6 Nottingham Health Profile domains were altered. When hepatocellular carcinoma patients were compared with matched cirrhotics, differences were present for Bodily Pain, Role Limitation-Physical, and the Physical Component Summary of Short Form-36, as well as Pain of Nottingham Health Profile. Perceived health status had changed significantly in the year prior to assessment. Health-related quality of life was not primarily related to tumour mass or hepatocellular failure, whereas sleep disorders were selected by logistic regression as strongly associated with poor health-related quality of life. CONCLUSIONS: The present data stress the relevance of pain in poor perceived health status of hepatocellular carcinoma patients, and the importance of minor symptoms, such as sleep disorders. PMID- 12725609 TI - Enalapril-induced acute recurrent pancreatitis. AB - Enalapril, an angiotensin-converting enzyme inhibitor, has several adverse effects, but acute pancreatitis is uncommon. The case of a patient with enalapril induced pancreatitis is described. Development of severe, necrotizing pancreatitis after inadvertent rechallenge confirmed the causal relationship between enalapril and acute pancreatitis. PMID- 12725610 TI - Pancreatic hyperenzymaemia and hypertransaminasaemia in healthy subjects. Report of three cases. AB - Three healthy subjects, two from Italy and one from the United States, showing a chronic increase in serum pancreatic enzymes and transaminases are described. The enzyme elevations reached very high levels but were not constant; rather, they fluctuated and sometimes returned to normal. Furthermore, tests for non-hepatic diseases that can be accompanied by an increase in serum transaminases, such as coeliac disease, were normal. The intervals between the first finding of the pancreatic hyperenzymaemia and the hypertransaminasaemia and this study ranged from 2 to 6 years (mean 4.3 years), during which the three subjects remained healthy, with no clinical, laboratory or imaging evidence of disease. These data support the conclusion that these increases in enzymes are benign; however, monitoring of these three subjects is already underway. Awareness of this anomaly is important, both to relieve the distress of the persons involved, as well as to avoid the numerous, sometimes invasive, complex and expensive examinations that might otherwise be unnecessarily performed. PMID- 12725611 TI - Unusual liver damage ensuing after warfarin administration in a pregnant woman with caval thrombosis. PMID- 12725612 TI - Validation of the accuracy of the H. pylori stool antigen test (HpSA) in the Japanese population. PMID- 12725613 TI - Celiac sprue: another autoimmune syndrome associated with hepatitis C? PMID- 12725614 TI - Review: oral protein and energy supplements reduce all-cause mortality in elderly persons. PMID- 12725615 TI - Dexamethasone improved disability in acute bacterial meningitis. PMID- 12725616 TI - Anterior-posterior electrode position led to more cardioversion success in atrial fibrillation. PMID- 12725617 TI - Intravenous water-soluble amiodarone improved 24-hour survival in incessant ventricular tachycardia. PMID- 12725618 TI - An Indo-Mediterranean diet was more effective than a control diet in primary and secondary coronary artery disease prevention. PMID- 12725619 TI - Review: concomitant aspirin use does not reduce the effectiveness of ACE inhibitors. PMID- 12725620 TI - Losartan did not differ from captopril for reducing all-cause mortality after acute myocardial infarction. PMID- 12725621 TI - Screening for abdominal aortic aneurysm reduced death from AAA in older men. PMID- 12725622 TI - 3 hyperbaric oxygen treatments reduced cognitive sequelae of acute carbon monoxide poisoning. PMID- 12725624 TI - Antimicrobial treatment did not reduce complications of urinary tract infection in diabetes and asymptomatic bacteriuria. PMID- 12725623 TI - Training in flexible intensive insulin management improved glycemic control and quality of life in type 1 diabetes. PMID- 12725625 TI - Omeprazole was effective in the short term for uninvestigated dyspepsia. PMID- 12725626 TI - Balsalazide achieved symptomatic remission sooner than mesalamine for ulcerative colitis. PMID- 12725627 TI - Review: oral or parenteral opioids alleviate dyspnea in palliative care. PMID- 12725628 TI - Review: ipratropium is not more effective than beta 2-agonists for acute exacerbations of chronic obstructive pulmonary disease. PMID- 12725629 TI - Review: terazosin improves urologic symptoms in benign prostatic hyperplasia. PMID- 12725630 TI - Self-administered behavioral training was equal to training plus biofeedback or verbal feedback in urge incontinence. PMID- 12725631 TI - Long-term mortality and cardiovascular morbidity increased in some patients with asymptomatic mitral valve prolapse. PMID- 12725632 TI - Long-term prognosis, measured by health status, disease process, and damage, varied in early rheumatoid arthritis. PMID- 12725633 TI - Review: elevated homocysteine levels are modestly associated with increased ischemic heart disease and stroke risk. PMID- 12725634 TI - Review: MTHFR TT genotype increases risk for coronary heart disease. PMID- 12725635 TI - Men and postmenopausal women with iron deficiency had increased risk for gastrointestinal malignancy. PMID- 12725636 TI - Activated protein C was cost-effective for prolonging survival in a subgroup of patients with severe sepsis. PMID- 12725637 TI - A gene expression profile independently predicted disease outcome in young women with breast cancer. PMID- 12725638 TI - Echocardiography was useful in evaluating unexplained syncope. PMID- 12725639 TI - Practice corner: clinical practice guidelines and handheld computers. PMID- 12725640 TI - Fetuin/alpha2-HS glycoprotein enhances phagocytosis of apoptotic cells and macropinocytosis by human macrophages. AB - Inflammatory diseases are associated with reduced serum concentrations of alpha(2)-HS glycoprotein (the human homologue of bovine fetuin), but the role of fetuin in inflammation is poorly understood. We hypothesized that fetuin may influence the resolution of inflammation by modulating the phagocytosis of apoptotic cells by macrophages. Using an in vitro flow cytometry-based phagocytosis assay, we investigated the role of fetuin in apoptotic cell clearance. Bovine fetuin and human alpha(2)-HS glycoprotein significantly augmented the phagocytosis of apoptotic cells by human peripheral blood monocyte derived macrophages, whereas the control proteins BSA, sialylated BSA and asialofetuin were ineffective. The enhancement of phagocytosis was concentration dependent, and required the presence of intact fetuin at the time of interaction between macrophages and apoptotic cells. Fetuin also substantially increased the uptake of labelled dextran 70000 by macrophages, which occurs by macropinocytosis, suggesting that this may be one of the mechanisms utilized for apoptotic cell uptake. PMID- 12725641 TI - Association of maternal and/or fetal factor V Leiden and G20210A prothrombin mutation with HELLP syndrome and intrauterine growth restriction. AB - This study was conducted to investigate the association of maternal and/or fetal factor V Leiden (FVL) and G20210A prothrombin mutation with HELLP syndrome. FVL and G20210A prothrombin mutation were determined using PCR. Sixty-three pregnant women, 36 of them diagnosed with HELLP syndrome, were included in the study. Overall, 68 children were born as a result of these pregnancies and blood sampling was possible in 28 out of 39 children from HELLP patients and 25 out of 29 children from the control women. The prevalence of a maternal FVL was elevated 2-fold in HELLP patients compared with the control women [six out of 36 (16.7%) compared with two out of 27 (7.4%); P =0.282]. None of the HELLP patients and only one woman in the control group was found to be positive for the G20210A prothrombin mutation (P =0.251). The fetal carrier frequency was four out of 28 compared with three out of 25 for FVL (P =0.811), and two out of 28 compared with one out of 25 for G20210A prothrombin mutation (P =0.629). Intrauterine growth restriction (IUGR) was significantly higher in fetuses found to be positive for a thrombophilic mutation (P =0.022). IUGR occurred in seven out of ten fetuses with a thrombophilic mutation compared with 11 out of 43 in fetuses without a mutation. The prevalence of FVL, but not of the G20210A prothrombin mutation, seems to be elevated in women with HELLP syndrome. A fetal thrombophilic mutation does not contribute significantly to the clinical features of the HELLP syndrome. Our results demonstrate a fetal contribution to IUGR. Fetal thrombophilic mutations may lead to placental microthrombosis, which consecutively could lead to a disturbed fetoplacental blood flow and thus cause growth restriction. PMID- 12725642 TI - Haemodynamic response to peripheral venous congestion in patients with unexplained recurrent syncope. AB - In patients with recurrent unexplained syncope, exaggerated peripheral venous pooling and impaired circulatory adjustment is thought to contribute to the outcome of a head-up tilt (HUT) test. The present study investigated the role of leg volume changes during venous congestion in the haemodynamic response of patients with recurrent unexplained syncope and the ability to predict the outcome of a HUT test. Changes in calf volume (strain gauge plethysmography), heart rate and arterial blood pressure were recorded in 60 patients with history of unexplained syncope (without postural tachycardia symptom) during venous congestion provoked by pneumatic thigh cuffs while supine at rest and during the initial 10 min of a 45 min 70 degrees HUT test. Twenty-seven patients [age (mean+/-S.D.), 39+/-16 years] exhibited symptoms [HUT(+)] and 33 patients (45+/ 14 years) were asymptomatic [HUT(-)]. During venous congestion, mean+/-S.E.M. calf volume increased in both groups [HUT(-), 4.5+/-0.2; HUT(+), 4.8+/-0.4 ml x 100 ml(-1), not significant), but significantly less during head-upright tilt [HUT(-), 3.3+/-0.2, P <0.01; HUT(+), 2.6+/-0.3 ml x 100 ml(-1), P <0.001] without differences between the groups. During venous congestion, arterial pressure increased significantly in asymptomatic HUT(-) patients, but not in the HUT(+) patients. Calf volume changes did not correlate with a symptomatic outcome to a 70 degrees HUT. The lack of exaggerated venous pooling during venous congestion and the inability of calf volume changes to predict a positive HUT suggest that excessive venous pooling does not contribute to the outcome of HUT. Attenuated changes in arterial pressure during venous congestion while supine suggest impaired adjustment of peripheral resistance to leg venous occlusion. PMID- 12725643 TI - Terminal vessel hyperperfusion despite organ hypoperfusion in familial dysautonomia. AB - Patients with familial dysautonomia (FD) exhibit orthostatic hypotension as well as recumbent hypertension. In addition, during dysautonomic crises, patients have hypertensive blood pressure that is presumed to be secondary to episodic vasoconstriction, as well as swollen hands that are presumed to be secondary to vasodilatation. This discrepancy in vascular control is poorly understood, yet may provide insight into the pathophysiology of autonomic crises. To evaluate the pathological mechanisms of overall blood flow and end-organ perfusion, we assessed resting and post-ischaemic limb and skin blood flow in FD patients. In groups of 15 FD patients and 15 controls, we measured resting and post-ischaemic forearm blood flow using venous occlusion plethysmography, and superficial skin blood flow using laser Doppler flowmetry. At rest, arterial inflow was averaged from eight venous occlusion measurements and expressed as percentage volume change/min. Post-ischaemic plethysmographic inflow was determined from the peak influx during the first venous occlusion following 3 min of ischaemia. Transcutaneous forearm partial pressures of oxygen and carbon dioxide were monitored continuously. At rest, plethysmographic limb perfusion was lower in FD patients than in controls, while skin blood flow did not differ between the two groups. After ischaemia, hyperperfusion of the forearm and hand was less pronounced in FD patients than in controls, while skin blood flow was significantly higher in patients than in controls. Partial pressures of O(2) and CO(2) did not differ between the two groups. We conclude that the reduced overall limb perfusion in patients with FD is due to hypertension-induced structural changes to vessel walls, with an increase in resistance vessel rigidity. The exaggerated post-ischaemic skin perfusion in FD patients seems to be due to deficient sympathetic innervation of precapillary vessels and arteriovenous shunts and to denervation hypersensitivity of intradermal small nerve fibres. Both the reduced limb perfusion and the dysfunctional end-organ blood supply in FD patients are likely to be major contributors to the vasomotor instability observed in these subjects, particularly during periods of stress. PMID- 12725644 TI - Up-regulation of vascular and renal mitogen-activated protein kinases in hypertensive rats is normalized by inhibitors of the Na+/Mg2+ exchanger. AB - In the present in vivo study, we have investigated whether inhibitors of the Na(+)/Mg(2+) exchanger quinidine and imipramine influence the development of hypertension and whether this is associated with modulation of mitogen-activated protein (MAP) kinase activation in arteries and kidneys of hypertensive rats. Sprague-Dawley rats were divided into four groups (n =6/group): control (vehicle), angiotensin II (Ang II; 150 ng/kg of body weight per min subcutaneously), quinidine [Ang II (150 ng/kg of body weight per min)+quinidine (5 mg/kg of body weight per day in food)] and imipramine groups [Ang II (150 ng/kg of body weight per min)+imipramine (5 mg/kg/day in food)]. Rats were studied for 3 weeks. Phosphorylation of vascular and renal extracellular-signal regulated protein kinase 1/2 (ERK1/2), p38MAP kinase and c-Jun N-terminal kinase (JNK) were assessed using phospho-specific antibodies. Ang II increased systolic blood pressure from 112+/-5 mmHg to 215+/-9 mmHg ( P <0.01). Development of hypertension was attenuated in Ang II-infused rats treated with quinidine (173+/ 6 mmHg) and imipramine (152+/-6 mmHg) (P <0.01). Phosphorylation of ERK1/2, p38MAP kinase and JNK, which were increased 2-3-fold in arteries of the Ang II group, were reduced by quinidine and imipramine (P <0.05). Activation of renal MAP kinases was also increased in the Ang II group (P <0.05). Quinidine and imipramine reduced the phosphorylation of renal ERK1/2, but did not modify renal p38MAP kinase or JNK. Our data demonstrate that Ang II induces severe hypertension in Sprague-Dawley rats and this is associated with increased phosphorylation of vascular and renal MAP kinases. Quinidine and imipramine attenuated the development of hypertension and normalized MAP kinase activity. The findings from this study suggest a possible role for the Na(+)/Mg(2+) exchanger in vascular signalling events associated with blood pressure elevation in Ang II-dependent hypertension. PMID- 12725647 TI - Using multiple survey vendors to collect health outcomes information: how accurate are the data? AB - BACKGROUND: To measure and assess health outcomes and quality of life at the national level, large-scale surveys using multiple vendors to gather health information is becoming the norm. This study evaluates the ability of multiple survey vendors to gather and report data collected as part of the 1998 Medicare Health Outcomes Survey (HOS). METHOD: Four hundred randomly sampled completed mailed surveys were chosen from each of six certified vendors (N = 2397) participating in the 1998 HOS. The accuracy of the data gathered from the vendors was measured by creating a "gold standard" record for each survey and comparing it to the final record submitted by the vendor. RESULTS: Overall rates of agreement were calculated, and ranged from 97.0% to 99.8% across the vendors. CONCLUSION: Researchers may be confident that using multiple vendors to gather health outcomes information will yield accurate data. PMID- 12725645 TI - Steroid-induced polycystic ovaries in rats: effect of electro-acupuncture on concentrations of endothelin-1 and nerve growth factor (NGF), and expression of NGF mRNA in the ovaries, the adrenal glands, and the central nervous system. AB - Previous studies on the effect of repeated electro-acupuncture (EA) treatments in rats with steriod-induced polycystic ovaries (PCO), EA has been shown to modulate nerve growth factor (NGF) concentration in the ovaries as well as corticotropin releasing factor (CRF) in the median eminence (ME). In the present study we tested the hypothesis that repeated EA treatments modulates sympathetic nerve activity in rats with PCO. This was done by analysing endothelin-1 (ET-1), a potent vasoconstrictor involved in ovarian functions, as well as NGF and NGF mRNA expression involved in the pathophysiological process underlying steroid-induced PCO. The main result in the present study was that concentrations of ET-1 in the ovaries were significantly lower in the PCO group receiving EA compared with the healthy control group (p < 0.05). In the hypothalamus, however, ET-1 concentrations were found to be significantly higher in the PCO group receiving EA than in the healthy control group (p < 0.05). Concentrations of ovarian NGF protein were significantly higher in the PCO control group compared with the healthy control group (p < 0.001), and these concentrations decreased significantly after repeated EA treatments compared with those in the PCO control group (p < 0.05) and were found to be the same as those in the healthy control group. In conclusion, these results indicate that EA modulates the neuroendocrinological state of the ovaries, most likely by modulating the sympathetic nerve activity in the ovaries, which may be a factor in the maintenance of steroid-induced PCO. PMID- 12725651 TI - Guidelines for using in vitro methods to study the effects of phyto-oestrogens on bone. AB - These guidelines review the relevant literature on the way plant phyto-oestrogens act on bone and the responsiveness of different bone cell systems to phyto oestrogenic compounds. The primary emphasis is on the experimental conditions used, the markers available for assessing osteoblast and osteoclast function, and their expected sensitivity. Finally, we assess the published results to derive some general recommendations for in vitro experiments in this area of research. PMID- 12725650 TI - Analysis of phyto-oestrogens in biological matrices. AB - A review covering different methods for the analysis of phyto-oestrogens in biological matrices is presented. Sample pretreatment and analysis of isoflavonoids and lignans by HPLC and GC with various detection methods are discussed. The immunoassay method is also briefly presented. PMID- 12725646 TI - Quantitative investigation of reproduction of gonosomal condensed chromatin during trophoblast cell polyploidization and endoreduplication in the East European field vole Microtus rossiaemeridionalis. AB - Simultaneous determinations of DNA content in cell nuclei and condensed chromatin bodies formed by heterochromatized regions of sex chromosomes (gonosomal chromatin bodies, GCB) have been performed in two trophoblast cell populations of the East-European field vole Microtus rossiaemeridionalis: in the proliferative population of trophoblast cells of the junctional zone of placenta and in the secondary giant trophoblast cells. One or two GCBs have been observed in trophoblast cell nuclei of all embryos studied (perhaps both male and female). In the proliferative trophoblast cell population characterized by low ploidy levels (2-16c) and in the highly polyploid population of secondary giant trophoblast cells (32-256c) the total DNA content in GCB increased proportionally to the ploidy level. In individual GCBs the DNA content also rose proportionally to the ploidy level in nuclei both with one and with two GCBs in both trophoblast cell populations. Some increase in percentage of nuclei with 2-3 GCBs was shown in nuclei of the placenta junctional zone; this may be accounted for by genome multiplication via uncompleted mitoses. In nuclei of the secondary giant trophoblast cells (16-256c) the number of GCBs did not exceed 2, and the fraction of nuclei with two GCBs did not increase, which suggests the polytene nature of sex chromosomes in these cells. In all classes of ploidy the DNA content in trophoblast cell nuclei with the single GCB was lower than in nuclei with two and more GCBs. This can indicate that the single GCB in many cases does not derive from fusion of two GCBs. The measurements in individual GCBs suggest that different heterochromatized regions of the X- and Y-chromosome may contribute in GCB formation. PMID- 12725652 TI - Phyto-oestrogen levels in foods: the design and construction of the VENUS database. AB - The objective of the Vegetal Estrogens in Nutrition and the Skeleton (VENUS) project was to evaluate existing data on dietary exposure to compounds with oestrogenic and anti-oestrogenic effects present in plant foods as constituents or contaminants, and to identify and disseminate in vitro and in vivo methodologies to analyse the effects of such compounds on bone. To permit the assessment of exposure to isoflavones in European populations (Italy, the UK, Ireland, The Netherlands), the VENUS database of phyto-oestrogen levels in foods was established. Data on the isoflavone (genistein and daidzein) content of 791 foods, including almost 300 foods commonly consumed in Europe, were collected. Levels of coumestrol, formononetin and biochanin A in a limited number of foods were also included. Lignan levels (secoisolariciresinol and matairesinol) in 158 foods were incorporated into the database, which also contains information on the references sourced for the compositional data, on the analytical methods used by each author and on the number of foods analysed in each reference. The VENUS database was constructed in Microsoft Access 2000, which is widely available as part of Microsoft Office Professional. This paper outlines the procedures used for the selection and evaluation of existing literature data for incorporation into the database. In addition, the design of the database is described, along with the data entry and quality control procedures used in its construction. Limitations of the data are discussed and guidelines for its use are provided. PMID- 12725653 TI - Isoflavone intake in four different European countries: the VENUS approach. AB - The aim of this study was to identify the level of isoflavone intake (total isoflavones, daidzein and genistein) in four European countries: Ireland, Italy, The Netherlands and the UK. For this purpose national food composition databases of isoflavone content were created in a comparable way, using the Vegetal Estrogens in Nutrition and the Skeleton (VENUS) analytical database as a common basis, and appropriate food consumption data were selected. The isoflavone intake in Ireland, Italy, The Netherlands and the UK is on average less than 1 mg/d. Small groups of consumers of soya foods could be identified in Ireland, The Netherlands and the UK. The estimated intake levels are low compared with those found in typical Asian diets (approximately 20-100 mg/d) and also low compared with levels where physiological effects are expected (60-100 mg/d). The results (including a subgroup analysis of soya product consumers) showed that such levels are difficult to achieve with the European diets studied here. PMID- 12725654 TI - Phyto-oestrogen database of foods and average intake in Finland. AB - Information on phyto-oestrogen intake in various populations has been scanty until now, primarily because data on the content of these compounds in foods were lacking. We report here on expansion of the Finnish National Food Composition Database (Fineli) with values for the plant lignans matairesinol and secoisolariciresinol and the isoflavones daidzein and genistein. The values, expressed as aglycones, were based on food analyses (mainly GC-MS) or imputed from analytical data for 180 foods for lignans and 160 foods for isoflavones; additionally, over 1000 values were derived from the recipe database of Fineli. Average intake of these phyto-oestrogens was calculated using food consumption data of the National Dietary Survey FINDIET 1997, which was carried out in a random sample of the adult population in five areas in Finland. The dietary data were collected by 24 h recall =2862). The mean lignan intake was 434 (standard deviation (SD) 1575) microg/d and the mean isoflavone intake was 788 (SD 673) microg/d. Women had a higher lignan density (microg lignans/MJ) in their diet than men (P<0.05). Men had a higher mean daily isoflavone intake, 902 (SD 368) microg, than women, 668 (SD 963) microg (P<0.05). The sources of lignans were many: seeds, cereals, fruit, berries and vegetables. The main sources of isoflavones appeared to be processed meat products/sausages containing soya as an ingredient, and legumes as such. The average intake of lignans and isoflavones in Finland seems to be low, but intake varies throughout the population. PMID- 12725655 TI - Food sources of phyto-oestrogens and their precursors in Europe. AB - Phyto-oestrogens are dietary components found in some plants, which act in vivo like weak oestrogens. They may reduce the risk of some degenerative diseases moderated by oestrogen, including breast cancer and osteoporosis. The most widely studied are the isoflavones genistein and daidzein from soyabeans, but lignans may be more prevalent in the European diet. Soya foods have traditionally been consumed in the Orient for millennia, and are now widely available to European consumers. Levels of isoflavone in soyabeans from published literature vary between 560 and 3810 mg/kg, depending on variety and growing conditions. Soya protein concentrates and isolates derived from soyabeans contain 466-615 mg isoflavones/kg. Traditional soya milk, bean curds, bean sprouts, etc. contain 13 to 2030 mg isoflavone/kg, depending on the starting raw material and final water content. Fermented foods have a different isoflavone conjugate profile, which may be important in absorption and metabolism. Soya analogues of European foods include dairy and meat products, which contain 38 to 3000 mg total isoflavones/kg, depending the source of soya and dilution with other ingredients. A wide range of foods contain low levels of soya-derived isoflavones, but such foods do not make a significant contribution to mean intakes in Europe. Flaxseed is by far the richest source of lignan precursors. However, foods such as cereal brans, legumes and some vegetables are a more important source in the diets of Europeans because they are more widely consumed. For similar reasons, compared with soya isoflavones, lignans may be a more important source of phyto-oestrogens in the diets of Europeans. PMID- 12725657 TI - Prevention of osteopaenia by phyto-oestrogens: animal studies. AB - Osteoporosis has become a major public health problem. Because the biggest culprit in the process of bone loss is oestrogen deficiency, hormone replacement therapy remains the mainstay for prevention, but prophylaxis by this means is limited. Phyto-oestrogens deserve special mention because emerging data support the suggestion that these weakly oestrogenic compounds, present in plants, may prevent bone loss associated with the menopause and thus represent a potential alternative therapy for a range of hormone-dependent conditions, including postmenopausal symptoms. A substantial body of work in animal models in the past few years has provided convincing data for significant improvements in bone mass and other endpoints following feeding with soya. Thus, phyto-oestrogens appear to have potential promise for maintaining or modestly improving bone mass of human subjects when consumed at optimal dosages. However, we must appreciate the limits of the information reached before extrapolating to man and we need to gather more data before health professionals can actively advocate the increased consumption of soya. Indeed, it will be important further to characterise the physiological effects of phyto-oestrogens and their margins of safety. PMID- 12725656 TI - Bioavailability of phyto-oestrogens. AB - The term phyto-oestrogen encompasses isoflavone compounds, such as genistein and daidzein, found predominantly in soya products and the lignans, such as matairesinol and secoisolariciresinol, found in many fruits, cereals and in flaxseed. There is evidence that they have potential health benefits in man particularly against hormone-dependent diseases such as breast and prostate cancers and osteoporosis. This has led to intense interest in their absorption and biotransformation in man. The metabolism of isoflavones and lignans in animals and man is complex and involves both mammalian and gut microbial processes. Isoflavones are present predominantly as glucosides in most commercially available soya products; there is evidence that they are not absorbed in this form and that their bioavailability requires initial hydrolysis of the sugar moiety by intestinal beta-glucosidases. After absorption, phyto oestrogens are reconjugated predominantly to glucuronic acid and to a lesser degree to sulphuric acid. Only a small portion of the free aglycone has been detected in blood, demonstrating that the rate of conjugation is high. There is extensive further metabolism of isoflavones (to equol and O-desmethylangolensin) and lignans (to enterodiol and enterolactone) by gut bacteria. In human subjects, even those on controlled diets, there is large interindividual variation in the metabolism of isoflavones and lignans, particularly in the production of the gut bacterial metabolite equol (from daidzein). Factors influencing absorption and metabolism of phyto-oestrogens include diet and gut microflora. PMID- 12725658 TI - Investigating the role of natural phyto-oestrogens on bone health in postmenopausal women. AB - Research on the bone effects of natural phyto-oestrogens after menopause is at a relatively early stage. Published studies are few, difficult to compare and often inconclusive, due in part to design weaknesses. Currently, many questions remain to be answered including to what extent a safe daily intake may prevent postmenopausal bone loss. These questions can only be addressed by conducting well-planned, randomised clinical trials that take into consideration present knowledge in the oestrogen, phyto-oestrogen and bone fields. This review is intended to provide hints for critical decision-making about the selection of subjects, type of intervention, suitable outcome measures and variables that need to be controlled. PMID- 12725659 TI - Phyto-oestrogens and osteoporosis: what is a safe dose? AB - Hormone replacement therapy (HRT) for preventing loss of bone following the menopause is utilised by only 8-10 % of possible users, largely due to a fear of increased risk of breast cancer. Plant oestrogen-like compounds (phyto oestrogens) have been proposed as an alternative to HRT to prevent osteoporosis. One class of phyto-oestrogens (the isoflavones) is found in soya foods and red clover. The food industry is developing a wide variety of new foods containing soya to substantially increase isoflavone intake, as well as extracting isoflavones from soya and clover to use as additives to non-soya foods. Pharmaceutical companies are also preparing isoflavone extracts to be used in pill form. In each case the targeted delivery is approximately 50 mg of isoflavones/d. Is this dose of isoflavones safe? In this review of the current literature, it is concluded that isoflavones consumed orally and in doses below 2 mg/kg body weight per d should be considered safe for most population groups. Whether these doses are sufficient to prevent osteoporosis is a separate matter. PMID- 12725660 TI - Measuring cognition in menopause research: a review of test use. AB - OBJECTIVE: To examine critically how various aspects of memory functions have been assessed in menopause-related research. METHOD: Twenty peer-reviewed articles on the menopause and cognitive functioning published between 1991 and 2000 were identified through a computerized Medline search. All studies utilized healthy non-demented women, and assessed specific aspects of cognitive functioning using standardized tests rather than self-reporting of perceived functioning. RESULTS: Measures used to assess attention and concentration, verbal memory, learning and verbal expression, visual memory, concept formation and reasoning have all been reviewed. Findings from studies assessing attention and concentration demonstrate inconsistent results with regard to hormone levels or menopausal status. Although five measures of verbal learning and memory have been considered, no articles published in the past decade have reported unequivocal results for any one measure. Only one study reviewed found significant differences in measures of verbal expression between hormone therapy users and non-users. In addition, no strong association between estrogen level and visual memory was identified in the studies reviewed. CONCLUSIONS: This careful analysis demonstrates that, within the studies of cognitive abilities of menopausal women reviewed in this article, a wide variety of tests have been used. However, the body of literature considered neither consistently nor conclusively demonstrates a clear effect of exogenous hormone therapy on memory. It is suggested that verbal memory, visual memory, concept formation and reasoning should be simultaneously investigated in future research, and that covariate measures of attention and concentration, verbal expression and reasoning are also included. In addition, within each specific domain, the psychometric properties of tests, the populations they were intended to assess and the specific aspects of cognitive function they were designed to address need to be more closely considered. PMID- 12725661 TI - Has psychosocial research on the menopause any clinical relevance? AB - The relevance of the findings of longitudinal studies, and other types of psychosocial studies, of general population samples of climacteric women to day to-day clinical practice is examined. It is shown that the findings of such studies can act as a useful guide to the assessment and treatment of menopausal women in a clinical setting. However, it is argued that the relevance of these studies to clinical practice could be even greater if certain clinically derived considerations were taken into account in the methodology and design of such studies, and therefore provide a more clinically relevant evidence base for the overall management of women at the time of the menopause. PMID- 12725662 TI - Vascular function of forearm microcirculation in postmenopausal women with type 2 diabetes: potential benefit of hormone replacement therapy? AB - OBJECTIVE: To study the potential effects of hormone replacement therapy (HRT) on vascular function of the peripheral circulation in postmenopausal women with type 2 diabetes. METHODS: Seventeen premenopausal women aged 47.4 +/- 4.4 years (PreM), 23 nondiabetic postmenopausal women aged 59.4 +/- 7.0 years (PM), 15 postmenopausal women with type 2 diabetes aged 60.3 +/- 7.2 years (PMD) and 12 postmenopausal women with type 2 diabetes using HRT aged 61.2 +/- 4.0 years (PMDHRT) were studied. Vascular function of the peripheral circulation was investigated by measuring hyperemic responses of the forearm microcirculation following an ischemic stimulus using venous occlusion, strain-gauge plethysmography. Fasting lipids, glucose and glycated hemoglobin (HbA1c) were also measured. RESULTS: Forearm vascular function was significantly impaired in the PMD group: maximal blood flow (ml/100 ml/min) was 15.23 +/- 8.19 vs. 28.21 +/ 12.30 (PreM), 23.62 +/- 6.62 (PM) and 23.37 +/- 5.78 (PMDHRT) (p = 0.004); flow debt (ml/100 ml) was 3.99 +/- 2.83 vs. 7.40 +/- 4.92 (PreM), 5.66 +/- 3.67 (PM) and 8.57 +/- 4.84 (PMDHRT) (p = 0.0018); vascular resistance (mmHg/ml/100 ml/min) was 11.90 +/- 9.02 vs. 5.04 +/- 2.41 (PreM), 5.55 +/- 2.69 (PM) and 5.96 +/- 1.88 (PMDHRT) (p = 0.003). Fasting lipids, HbA1c and body mass index were not significantly different between the two diabetic groups. CONCLUSIONS: Postmenopausal women with diabetes have abnormal vascular function of resistance arteries that may improve with HRT. HRT may therefore reduce cardiovascular risk in postmenopausal women with type 2 diabetes, and a clinical end-point trial is warranted to test this hypothesis. PMID- 12725663 TI - Effects of transdermal estrogens on endothelial function in postmenopausal women with coronary disease. AB - OBJECTIVE: Oral estrogens improve endothelial function, and for this reason may be considered cardioprotective; however, in women with coronary heart disease there may also be an increase in the risk of thrombosis. Although transdermal estrogen administration may decrease this adverse effect, there are few data on endothelial function in women with coronary heart disease treated using such therapy. This study aimed to report the endothelial response in postmenopausal women with coronary heart disease treated with transdermal estrogen. MATERIALS AND METHODS: This was a double-blind, prospective, randomized study. Eighteen patients with a history of acute coronary syndrome and nine healthy women were studied over 4 weeks. Coronary patients were assigned at random to receive a patch containing either 50 microg estradiol or placebo on a weekly basis. Endothelial function was assessed by flow-mediated vasodilatation of the brachial artery. Baseline blood flow (brachial artery diameter) was measured after 30 min rest and following ischemia, prior to treatment and after 4 weeks. RESULTS: Flow mediated vasodilatation in normal patients was 17.8%, whereas in women with coronary disease it was 1.2% (p = 0.0001). Arterial diameter for the resting period in coronary disease subjects increased from 4.22 +/- 0.59 to 4.41 +/- 0.56 mm (p < 0.004) after 4 weeks of estrogen therapy, whereas, in women receiving placebo, it did not change. Flow-mediated vasodilatation in the estrogen group was 3.4% and in the placebo group was 0.5% (p = 0.05). CONCLUSIONS: Transdermal estrogen may improve endothelial function in women with coronary heart disease. PMID- 12725664 TI - A cohort study of topical vaginal estrogen therapy in women previously treated for breast cancer. AB - OBJECTIVE: To estimate the risk of recurrence of breast cancer associated with the use of topical vaginal estrogen therapy in the management of vaginal atrophy in women previously treated for breast cancer. METHODS: The study group comprised 1472 women with histologically confirmed breast cancer. In 69 of these subjects (4.7%) their only bothersome menopausal problems were vaginal symptoms. In these women, poorly absorbed topical vaginal estrogen cream or tablets were used. The response of these patients was compared with that of the rest of the database. A Cox regression analysis was performed using sex hormone usage after diagnosis as a time-dependent covariate. Disease-free interval was the outcome measured. Results are expressed as a hazard ratio with 95% confidence intervals. The hazard rate is defined as the probability of disease recurrence or of a subject dying from breast cancer over the study period. A second analysis was performed adjusting for factors known to affect breast cancer prognosis. RESULTS: Hormone usage was entered as a time-dependent covariate with disease-free interval as the outcome. Subjects who used a topical estrogen alone for menopausal symptoms had an uncorrected hazard ratio of 0.30 (95% confidence interval (CI) 0.11-0.80, p = 0.02). The corrected hazard ratio was 0.57 (95% CI 0.20-1.58, p = 0.28). The hazard rate for a subject dying was not analyzed, as there were too few numbers. CONCLUSIONS: Although the small numbers of this study preclude a definitive result, topical estrogen usage does not appear to be associated with an increased risk of recurrence of breast cancer. PMID- 12725665 TI - Relationship between climacteric symptoms and serum serotonin levels in postmenopausal women. AB - OBJECTIVE: The aim of the study was to evaluate the possible relationship between climacteric symptoms and serum serotonin concentrations in postmenopausal women. MATERIALS AND METHODS: We studied 39 women who attended the Department of Gynecological Endocrinology in Poznan because of climacteric complaints. We evaluated climacteric symptom severity with the use of the Kupperman scale and we measured 17beta-estradiol, follicle stimulating hormone (FSH) and serotonin serum concentrations by radioimmunoassay. RESULTS: The mean serum serotonin concentration in the studied women was 48.9 ng/ml (standard deviation, SD +/- 24.1 ng/ml). The mean serum serotonin concentration in the group of patients with mild climacteric symptoms was 97.7 ng/ml (SD +/- 16.6 ng/ml), and in the group of patients with moderate and severe climacteric symptoms was 36.2 ng/ml (SD +/- 24.1 ng/ml). The difference between the two groups was statistically significant (Mann-Whitney test, p < 0.05). There was no difference in serum serotonin concentrations between patients with or without specific symptoms (Mann-Whitney test, p > 0.05). CONCLUSIONS: Serum serotonin concentrations in postmenopausal women are related to the severity of climacteric symptoms. PMID- 12725666 TI - Continuation of hormone replacement therapy after hysterectomy. AB - OBJECTIVES: To assess long-term hormone replacement therapy (HRT) usage in women after hysterectomy and to assess the relationship between age and long-term use of HRT in these women. Problems and comments of those women responding to a questionnaire were evaluated. MATERIAL AND METHODS: A postal semistructured questionnaire survey was performed in a single gynecological practice. A total of 545 consecutive women who had undergone a hysterectomy for benign conditions between January 1986 and September 1997 were studied, the main outcome measure being continuing use of HRT. RESULTS: There was a response rate of 83.1% to the questionnaire; 83% of all responders were taking HRT at the time of the survey. A continuation rate of 95.7% was found among women who had had a hysterectomy after 1994, and of 84.7% among those operated on in 1989 or before. Implants were used by 68%, transdermal patches by 17%, oral preparations by 11% and estradiol gel by 4%. Ten per cent of those not taking HRT at present indicated that they were likely to start again in the near future. No correlation was found between age and likelihood of HRT continuation. Fifty per cent of women responding to the questionnaire made further comments: 17.6% of these made specific positive comments regarding HRT, 16.7% reported weight gain, 9.7% suffered breast symptoms and 13.2% admitted concerns regarding breast cancer. CONCLUSIONS: A high HRT continuation rate of between 95.7% (women having had their hysterectomy less than 5 years ago) and 84.7% (women 10 or more years from their operation) can be achieved in the long term. Considering the high proportion having implant therapy, the use of testosterone as well as estradiol replacement may be a major factor in the greater adherence to HRT of this group. PMID- 12725668 TI - Potential risks of phytoestrogens: experience from animal models. PMID- 12725667 TI - Use of hormone replacement therapy by menopausal women in six family-practice teaching clinics in Israel. AB - BACKGROUND: The necessity of hormone replacement therapy (HRT) remains a controversial subject, but guidelines suggest that the subject be discussed with all menopausal women. AIM: To determine the rate of HRT use among women cared for in family-practice, university-affiliated teaching clinics. METHODS: Physicians from six family-practice teaching clinics enrolled all menopausal patients, aged between 50 and 70 years, registered in their practice. A detailed questionnaire was completed regarding demographic and health variables, gynecological history and the nature of HRT use. RESULTS: In total, 682 women were invited to participate in the study; 587 (86%) participated. Some 74% of the women were married, 64% were secular and 43% stated that they performed regular exercise. Current HRT use was reported by 156 women (26.6%). Most women (83%) had been treated for over 1 year. Common indications for starting treatment were hot flushes (75%), and routine recommendation by their physician (68%). Eighty-three (14%) women were past HRT users. It was stated by 45% of never-users that HRT had never been offered to them. Current users of HRT were found to be younger than non-users (mean age 58.2 years vs. 61.5 years, p < 0.0001), more secular than traditional or religious (p < 0.0001) with more years of formal education (p < 0.0001), and more likely to be of Western or European ("Ashkenazi") origin (p < 0.0001). Current HRT users reported fewer chronic illnesses than did non-users (p = 0.001); they were more likely to report being engaged in regular exercise (p < 0.002) and were less likely to be obese (p < 0.005). In a multiple regression model, HRT use was associated with more years of formal education and a secular identity. CONCLUSION: Our study found a higher proportion of women using HRT than in other Israeli studies. HRT use was more prevalent among women with improved health status and higher socioeconomic status. The most common reason given for not using HRT was that a physician had not raised the subject, indicating that physician-related factors might contribute to using HRT. Thus, comparing HRT prescribing rates in non-teaching clinics should clarify this factor. PMID- 12725669 TI - Increased cesarean section rates in Turkey. AB - The aim of this study was to examine the trend in Cesarean section deliveries and the factors associated with Cesarean sections in Turkey. Data come from the ever married women questionnaire of the 1998 Turkish Demographic and Health Survey (TDHS-98). During the decade preceding the TDHS-98, the proportion of deliveries by Cesarean section increased from 5.7% to 20.8%. When only hospital births were considered, the percentage of Cesarean deliveries for the year 1998 was found to be 26.1%. The estimated rate for the year 2001 was around 30% (i.e. double the maximum rate of Cesarean sections defined by the World Health Organization). Logistic regression analysis performed for the births occurring in the most recent period of 1993-98 revealed that the highest Cesarean section rate was strongly associated with maternal education, maternal age, place of delivery, number with prenatal care and household welfare. These findings imply that women with higher socioeconomic status are more likely to accept Cesarean section than women with lower socioeconomic status. The trend of increasing Cesarean section rates is a problem in itself, but more importantly it may indicate that Turkey is headed toward a more costly medical delivery system. For all of these reasons, the reduction of Cesarean section rates should be a priority for any reproductive health program in Turkey in order to improve the quality of prenatal care and to reduce the number of maternal deaths and morbidity. PMID- 12725670 TI - Reasons for not using family planning methods in Eastern Turkey. AB - OBJECTIVE: The reasons for not using family planning methods among women aged 15 45 years were investigated in the Eastern region of Turkey. METHODS: A total of 518 randomly selected women were interviewed with a questionnaire. Information was collected on women's fertility behavior and intentions, sources of knowledge and practice of contraception and sociodemographic characteristics. RESULTS: The rate of women who were not using any form of family planning was 45.2%. The reasons for not using any form of family planning were: not having the approval of the husband or family leaders (38%); believing that it was a sin to use a family planning method (32.5%); believing that it could cause abnormal bleeding (14.1%); believing that it could cause infertility (7.3%); believing that it could cause cancer (4.7%); and believing that it could cause pelvic pain (3.4%). CONCLUSION: Efforts should be directed towards education of family members and religious leaders to increase the usage of family planning methods. PMID- 12725671 TI - Should endometrial hyperplasia be regarded as a reason for abnormal uterine bleeding in users of the intrauterine contraceptive device? AB - OBJECTIVE: Histopathological evaluation of the endometrium in cases having an intrauterine device (IUD) removed for abnormal uterine bleeding. METHODS: Fifty eight consecutive patients with a complaint of uterine bleeding leading to IUD removal were recruited for this study. Endometrial sampling, via Novak curette, was performed during IUD removal for histopathological evaluation of the endometrium. RESULTS: A total of 58 current IUD users, presenting with uterine bleeding, were retrospectively analyzed. Mean age of the women was 37.1 +/- 2.1 years (range 25-43). Mean duration of IUD use was 4.2 +/- 1.3 years (range 1-10). Out of 58 cases, 21 (36.2%) did not harbor any endometrial pathology. In eight cases (13.8%), hyperplastic endometrial changes were apparent, six of which were simple hyperplasia (four of them were focal). Two cases of complex hyperplasia were detected (one was diffuse with atypia and one was focal without atypia). In this series, there was one case with an endometrial polyp. Mean age of cases with hyperplastic endometrium was found to be statistically high compared to those with normal endometrial histology (42.4 +/- 3.2 vs. 37.6 +/- 2.1, p = 0.04). In cases with IUD use of > 5 years, chronic endometritis was more prevalent, compared to those with < 5 years of use (chi(2) 5.08, p = 0.02). CONCLUSIONS: IUD use is a risk factor for chronic endometritis. Nevertheless, in 13.8% of cases in this series, as a reason for abnormal uterine bleeding, other than endometritis, endometrial hyperplasia constituted the second most common endometrial pathology among cases over the age of 40, on current IUD use and having complaints of abnormal uterine bleeding. Hence, this finding should prompt the physician to perform endometrial sampling in users of an IUD over the age of 40, presenting with abnormal uterine bleeding. PMID- 12725672 TI - The use of family planning methods in an administrative district of Istanbul. AB - In Turkey, the first population and family planning law was accepted by the government in 1965, and this law was revised in 1982. This study was carried out to evaluate the family planning methods of women who applied to rural health centers of Kucukcekmece and were willing to use the health-care service and consultancy about birth control methods. The data of the study were gathered by means of an appropriate inquiry form and face-to-face interview with the women willing to use the family planning service between September 1998 and June 1999 at the health-care centers of Kucukcekmece of Istanbul. There were 699 women participants in the study. The last pregnancy of 27.2% (n = 175) of the women had been undesired; 86.8% of all the women who participated in the study declared that they did not desire any more children, but only 81.3% (n = 568) out of these were using a method to protect from pregnancy. The most frequently known (84.8%) and used (45.9%) method of the participants was an intrauterine device (IUD), and 80.8% of the women using an IUD wanted to continue using that method. A total of 223 women (39.3%) using any method protecting from pregnancy became pregnant. Approximately half (57.8%) of those occurred during the use of withdrawal. According to the findings of this study, the level of knowledge about family planning and the consequent behavior of the women within the fertile age group are better than the average of Turkey, but still not satisfactory. PMID- 12725673 TI - Implanon: user views in the first year across three family planning services in the Trent Region, UK. AB - OBJECTIVE: Implanon (NV Organon, Oss, The Netherlands) was released in the United Kingdom towards the end of 1999. This survey elicited the experiences of women who had the new implant inserted in three family planning services in North Trent, UK during the first year after its introduction. METHOD: A previously piloted questionnaire was sent to all Implanon users who had the devices fitted in the three services between December 1999 and December 2000. Returned questionnaires were analyzed. RESULTS: Health-care providers and friends or family were the main sources of information about the single-rod contraceptive implant for the survey population. Women considered themselves informed about the method prior to insertion irrespective of whether they continued to use the method or requested early removal. Ease of use was the most common reason for choosing Implanon and one of the best-liked features. Bleeding irregularities were the most commonly reported side-effect, followed by weight gain, moods and headaches. Bleeding problems led to the majority of removal requests; the wish for pregnancy was not stated at all. The single-rod implant compared favorably with most responders' experiences with other methods of contraception. Further research into motivation and perceptions regarding modern contraceptives may be required in different populations. PMID- 12725674 TI - Effect of an oral contraceptive containing drospirenone and ethinylestradiol on general well-being and fluid-related symptoms. AB - Oral contraception is the most widely used reversible contraceptive method. Continuous research over the past decades has led to a range of highly reliable, effective and safe oral contraceptives. Newly developed progestogens may also provide additional non-contraceptive health-related benefits that differentiate the products from each other. Women desiring contraception may thus choose from a wide range of oral contraceptives according to their individual needs. A variety of physical and emotional changes have been linked to hormonal fluctuations during the menstrual cycle. To date, only very few studies have been performed on the impact of fluid retention-related symptoms on well-being and few data are hence available on suggested methods of measurement. This open, multicenter, uncontrolled study evaluated the effects of a combined preparation containing 3 mg drospirenone and 30 microg ethinylestradiol (Yasmin, Schering AG, Berlin, Germany) on general well-being and fluid-related symptoms in women experiencing psychological, behavioral and somatic premenstrual symptoms. The study was conducted over six 28-day cycles, with 336 subjects enrolled. A significant beneficial effect on psychological general well-being, as measured by the Psychological General Well-Being Index (PGWBI), was evident by cycle 3 and maintained at cycle 6. There was a significant reduction in both the incidence and severity of somatic symptoms associated with the menstrual cycle (abdominal bloating and breast tension) during treatment. Assessment by the investigator showed that 80% of subjects had improved on study treatment and 75% of subjects considered themselves satisfied with the study treatment. There was good agreement between the clinician and subject in their assessment of the treatment. Cycle control was very good and body weight remained stable or decreased slightly during the study. In conclusion, 3 mg drospirenone in combination with 30 microg ethinylestradiol has been shown to have a beneficial effect on psychological general well-being, as measured by the PGWBI. Reductions in the incidence and severity of somatic symptoms associated with the menstrual cycle were also observed, suggesting a beneficial effect due to the antimineralocorticoid nature of drospirenone. To our knowledge, this is the first study on oral contraceptives which has used the PGWBI in this population. As quality of life is one of the least explored segments in oral contraceptive users, more studies should investigate the impact of oral contraceptives on quality of life and general well being in this overall healthy population. PMID- 12725675 TI - High pregnancy rates and reproductive health indicators among female injection drug users in Vancouver, Canada. AB - OBJECTIVE: To determine the incidence of pregnancy among active injection-drug users and to identify factors associated with becoming pregnant. METHODS: The Vancouver Injection Drug User Study (VIDUS) is a prospective cohort study that began in 1996. Women who had completed a baseline and at least one follow-up questionnaire between June 1996 and January 2002 were included in the study. Parametric and non-parametric methods were used to compare characteristics of women who reported pregnancy over the study period with those who did not over the same time period. RESULTS: A total of 104 women reported a primary pregnancy over the study period. The incidence of pregnancy over the follow-up period was 6.46 (95% confidence interval (CI) 5.24-7.87) per 100 person-years. The average age of women who reported pregnancy was younger than that of women who did not report pregnancy (27 vs. 32 years, p < 0.001). Women of Aboriginal ethnicity were more likely to report pregnancy (odds ratio 1.6, 95% CI 1.0-2.5). Comparison of drug use showed no significant differences in pregnancy rate with respect to the use of heroin, cocaine or crack (p > 0.05). In examining sexual behavior, women who reported having had a regular partner in the previous 6 months were three times more likely to have reported pregnancy. Despite the fact that 67% of women in this study reported using some form of contraception, the use of reliable birth control was low. Only 5% of women in our study reported the use of hormonal contraceptives. CONCLUSION: There were a high number of pregnancies among high risk women in this cohort. This corresponded with very low uptake of reliable contraception. Innovative strategies to provide reproductive health services to at-risk women who are injecting drugs is a public health priority. PMID- 12725679 TI - High-dimensional analysis of evolutionary autonomous agents. AB - This article presents a new approach to the important challenge of localizing function in a neurocontroller. The approach is based on the basic functional contribution analysis (FCA) presented earlier, which assigns contribution values to the elements of the network, such that the ability to predict the network's performance in response to multi-unit lesions is maximized. These contribution values quantify the importance of each element to the tasks the agent performs. Here we present a generalization of the basic FCA to high-dimensional analysis, using high-order compound elements. Such elements are composed of conjunctions of simple elements. Their usage enables the explicit expression of sets of neurons or synapses whose contributions are interdependent, a prerequisite for localizing the function of complex neurocontrollers. High-dimensional FCA is shown to significantly improve on the accuracy of the basic analysis, to provide new insights concerning the main subsets of simple elements in the network that interact in a complex nonlinear manner, and to systematically reveal the types of interactions that characterize the evolved neurocontroller. PMID- 12725681 TI - Toward a formalization of emergence. AB - Emergence is a concept widely used in the sciences, the arts, and engineering. Some effort has been made to formalize it, but it is used in various contexts with different meanings, and a unified theory of emergence is still distant. The ultimate goal of a theory of emergence should include using emergence to model, design, or predict the behavior of multiagent systems. The author proposes a formal definition of a basic type of emergence using a language-theoretic and grammar systems approach. It is shown which types of phenomena can be modeled in this sense and what the consequences are for other more complex phenomena. PMID- 12725680 TI - Self-replicating machines in continuous space with virtual physics. AB - JohnnyVon is an implementation of self-replicating machines in continuous two dimensional space. Two types of particles drift about in a virtual liquid. The particles are automata with discrete internal states but continuous external relationships. Their internal states are governed by finite state machines, but their external relationships are governed by a simulated physics that includes Brownian motion, viscosity, and springlike attractive and repulsive forces. The particles can be assembled into patterns that can encode arbitrary strings of bits. We demonstrate that, if an arbitrary seed pattern is put in a soup of separate individual particles, the pattern will replicate by assembling the individual particles into copies of itself. We also show that, given sufficient time, a soup of separate individual particles will eventually spontaneously form self-replicating patterns. We discuss the implications of JohnnyVon for research in nanotechnology, theoretical biology, and artificial life. PMID- 12725682 TI - RandScape: complex images from simple algorithms. AB - This letter presents a range of complex patterns, some that seem eerily lifelike, that were produced from some simple algorithms. Some observations are offered that suggest the algorithms are modeling some natural processes. PMID- 12725683 TI - Artifact & artifice: views on life. AB - The views of some artists on what constitutes life are explored, with the aim of challenging those within the artificial life research community to rethink and perhaps expand their own views about the term and its meaningful application. The focus is on the musical works of Steve Reich and the paintings of Wassily Kandinsky. The role of the observer in determining when it is appropriate to label a thing as living is also discussed. PMID- 12725684 TI - The evolution of antigen-specific CD8+ T cell responses after natural primary infection of humans with Epstein-Barr virus. AB - Epstein-Barr virus (EBV) is a persistent, gamma-herpes virus that infects 90% of the human population. Primary infection, particularly if it is delayed until adolescence or beyond, may cause acute infectious mononucleosis and persistent infection may be associated with the development of several malignancies. CD8(+) T cells play a critical role in controlling both the primary and persistent phases of infection. This review summarises work that has been done characterising the primary immune responses to EBV. It goes on to describe the down regulation of the primary immune response and to discuss some of the factors that may be involved in determining the death or survival of populations of antigen-specific CD8(+) T cells. Finally it describes features of the populations of memory cells that mediate the long-term control of EBV in healthy seropositive individuals. The studies show differences in the responses to epitopes from lytic cycle versus latent proteins and highlight the complexity of naturally occurring, in vivo, immune responses. A clear understanding of the means by which CD8(+) T cells control EBV is important if we are to successfully develop vaccines and other forms of immunotherapy for the virus and its related malignancies. PMID- 12725685 TI - Possible mechanisms of protection elicited by candidate rotavirus vaccines as determined with the adult mouse model. AB - Rotaviruses cause extensive morbidity and mortality worldwide each year, supporting the need for a vaccine that is effective against rotavirus disease in all socioeconomic environments. Vaccines evaluated in clinical trials have all been live viruses that are delivered orally to mimic the excellent protection against severe rotavirus disease consistently observed after natural infection. The mechanisms by which either these vaccine candidates or natural rotavirus infections elicit protection are poorly understood. Therefore, it is not surprising that several of these candidate vaccines have provided little or no protection and have been discontinued. Two candidate vaccines are presently in phase III trials. These two were developed on the basis of very different views regarding the importance of one specific immune effector, that is, serotype specific neutralizing antibody. One of these candidates (RotaTeq) is composed of five bovine/human reassortant rotavirus strains containing neutralization proteins representative of dominant human serotypes. The other candidate (Rotarix) is composed of only a single strain of human rotavirus. Very recent data obtained with Rotarix support the suggestion that factors other than neutralizing antibody can play important roles in protection against rotavirus disease after live rotavirus immunization. These results must be confirmed in subsequent studies in different locales with circulating rotaviruses belonging to a variety of serotypes in order to establish there overall applicability. Mechanisms by which rotavirus immunization with live viruses or other immunogens elicit protection have been most extensively examined in an adult mouse model and were reported to be multi-factorial. That is, CD8 and CD4 T cells as well as B cells were all found to play significant roles. The importance of each lymphocyte population as effectors of protection was found to be dependent on the immunogen and the route of immunization. The results of studies in the adult mouse model that appear to be most pertinent to the present vaccine candidates are presented and discussed. PMID- 12725686 TI - The complex relationship between respiratory syncytial virus and allergy in lung disease. AB - Epidemiologic studies suggest a strong link between severe respiratory syncytial virus (RSV)-induced bronchiolitis in infancy and allergic disease; however, the mechanisms determining this relationship are currently unknown. In this review article, we summarize data from human clinical studies that explore the association between RSV infection and allergy, some that suggest that RSV bronchiolitis requiring hospitalization leads to an increased incidence of the allergic phenotype and others that suggest that pre-existing allergy is a risk factor for severe RSV bronchiolitis. We also review the published murine models that combine RSV infection and allergic sensitization that attempt to explain the complex relationship between these two factors in regard to lung immunopathology and physiologic dysfunction. PMID- 12725687 TI - Simian/human immunodeficiency virus(89.6) expressing the chemokine genes MIP 1alpha, RANTES, or lymphotactin. AB - We constructed replication competent, attenuated, nef-deleted SHIV(89.6) that express the rhesus macaque chemokine genes MIP-1alpha, RANTES, or LTN from the nef region. The chemokine inserts were stable during several passages in CEMx174 cells and the viruses grew well in activated rhesus PBMC. Expression of virally encoded MIP-1alpha, RANTES, or LTN was detected in culture fluids from infected HOS CD4(+) CXCR4(+) cells, that were used because they have a low background production of these chemokines. The in vitro growth kinetics of all nef-deleted SHIV(89.6) were slower than the parental strain in both CEMx174 cells and rhesus PBMC. Rhesus macaques were susceptible to SHIV(89.6-MIP-1alpha), SHIV(89.6 RANTES), SHIV(89.6-LTN), and nef-deleted control SHIV(89.6-dLTN) infection via the intrarectal route using standard virus doses, and intact viruses were reisolated from infected animals throughout the interval of acute infection. SHIV expressing the chemokine genes MIP-1alpha, RANTES, or LTN may help determine the in vivo roles for these chemokines in modulating virus replication and disease. PMID- 12725688 TI - Modulation of immune system function by measles virus infection. II. Infection of B cells leads to the production of a soluble factor that arrests uninfected B cells in G0/G1. AB - Measles can result in a variety of immunologic defects. Previously we showed that an Epstein-Barr virus-transformed B cell line (B cells), when infected with measles virus, produced a soluble antiproliferative factor that inhibited proliferation of T and B cells. Here we explore the effects of infection by measles virus versus the virus-free soluble antiproliferative factor on B cells. The B cells showed no change in the amounts of interleukin (IL)-2, 10, 12, interferon (IFN)-gamma, or transforming growth factor (TGF)-beta when infected or exposed to the soluble factor. Similarly, B cells showed no change in the expression of class II major histocompatibility antigens, LFA-1, ICAM-1, CD19, CD40, CD80, CD86, CD95 (Fas), or CD178 (FasL). Cell cycle analysis showed that measles virus infection caused an accumulation of cells in S and G(2)/M phases with a "sub-G(1)" cell population, while incubation of cells with the soluble factor caused an accumulation in G(0)/G(1). These experiments provide evidence that measles virus causes a profound inhibition of B cell proliferation without distinguishable changes in cytokine profile or cell surface phenotype. Further, it appears that there are two populations of cells affected by infection: one population is growth arrested due to the influence of the immunosuppressive factor and is not infected; a second population that is infected progresses through S phase less efficiently. Alternatively, while both the soluble factor and live virus infection may affect cells in G(0)/G(1) phases, only live virus infection could selectively induce apoptosis of G(0)/G(1) cells, resulting in cell accumulation in S and G(2)/M phases with a build up of "sub-G(1)" cells. PMID- 12725690 TI - Antibody-dependent enhancement of virus infection and disease. AB - In general, virus-specific antibodies are considered antiviral and play an important role in the control of virus infections in a number of ways. However, in some instances, the presence of specific antibodies can be beneficial to the virus. This activity is known as antibody-dependent enhancement (ADE) of virus infection. The ADE of virus infection is a phenomenon in which virus-specific antibodies enhance the entry of virus, and in some cases the replication of virus, into monocytes/macrophages and granulocytic cells through interaction with Fc and/or complement receptors. This phenomenon has been reported in vitro and in vivo for viruses representing numerous families and genera of public health and veterinary importance. These viruses share some common features such as preferential replication in macrophages, ability to establish persistence, and antigenic diversity. For some viruses, ADE of infection has become a great concern to disease control by vaccination. Consequently, numerous approaches have been made to the development of vaccines with minimum or no risk for ADE. Identification of viral epitopes associated with ADE or neutralization is important for this purpose. In addition, clear understanding of the cellular events after virus entry through ADE has become crucial for developing efficient intervention. However, the mechanisms of ADE still remain to be better understood. PMID- 12725689 TI - Experimental infection of ponies with equine influenza A2 (H3N8) virus strains of different pathogenicity elicits varying interferon and interleukin-6 responses. AB - The production of interferon (IFN), interleukin-6 (IL-6), and tumor necrosis factor (TNF) was monitored in horses during the course of influenza A2 virus infections. The effects of two virus strains, Newmarket/2/93 and Sussex/89, were compared, of which the latter is considered the more pathogenic in terms of clinical signs. Ten naive ponies were infected with influenza A/equine/Sussex/89 and 10 with influenza A/equine/Newmarket/2/93, respectively. As expected ponies infected with Sussex/89 showed the most pronounced clinical signs but there was no notable difference in viral excretion compared with Newmarket/2/93. IFN was detected in nasal secretions of all ponies infected with Sussex/89 but only in 2 ponies infected with Newmarktet/2/93. IFN was not detected in serum of any animal. IL-6 activity was detected in nasal secretions of all experimental animals from day 2 and onwards, but showed markedly higher IL-6 responses were observed in ponies infected with Sussex/89. No TNF activity was detected in any of the samples collected. In summary, equine influenza A 2 infections elicited local, and in some cases systemic, IFN and IL-6 responses in the ponies. Interestingly, there was some evidence that the duration and levels of cytokine responses may be related to the pathogenicity of the influenza strains. PMID- 12725692 TI - Preparation of samples of plant material for chromatographic analysis. AB - The selection, collection, and preliminary treatment of plant material samples are discussed. Stages of sample preparation and extraction techniques presently used are described, and the most recent examples of sample preparation for chromatographic analysis are reviewed. Recent applications of gas chromatography for the assessment of emission of volatile organic compounds by plants are also described. PMID- 12725691 TI - PKR is not required for interferon-gamma inhibition of VSV replication in neurons. AB - In this report, the contribution of PKR to the IFN-gamma mediated inhibition of VSV replication in neurons was examined. IFN-gamma treatment of NB41A3 murine neuroblastoma cells resulted in the reduced expression of VSV protein during infection. PKR was found to be modestly upregulated in NB41A3 cells following IFN gamma treatment. The phosphorylation state of PKR and its downstream target, eIF2alpha, were unaffected by either IFN-gamma or VSV infection. Inhibition of PKR through the use of 2-aminopurine or the expression of the Influenza A NS1 gene had no effect on the ability of IFN-gamma to inhibit the replication of VSV in vitro. These data indicate that endogenously expressed PKR is not required for the IFN-gamma mediated inhibition of VSV replication in NB41A3 neuroblastoma cells. PMID- 12725693 TI - Chromatographic behavior of ion pair enantiomers of dansyl leucine cyclohexylammonium salt on a beta-cyclodextrin stationary phase and the effect of a competitive-binding mobile phase additive. AB - The separation of dansyl leucine enantiomers on a beta-cyclodextrin stationary phase is significantly complicated by the association of the amino acid with its cyclohexylammonium counter ion, in a mobile phase of 80:20 (v/v) methanol-water. This produces very unusual chromatography, with two partially superimposed peaks observed for each enantiomer at lower column temperatures. The peak shape is attributed to the irreversible, oncolumn conversion of the ion pair (I) to the free, protonated (neutral) dansyl amino acid (II+H). Increasing the ionic strength of the mobile phase greatly improves the chromatography by transforming the solute species to enantiomers of II (the anionic, free amino acid). Van't Hoff plots are constructed for both species I and II (under different mobile phase conditions) to provide thermodynamic insight into the major enantioselective driving forces of separation. The chiral discrimination of the stationary phase is found to be primarily enthalpically driven for both solutes. Finally, 1-adamantanecarboxylic acid (ACA) is investigated as a solute competitive mobile phase additive to intentionally block the hydrophobic cyclodextrin cavities on the stationary phase. By varying the concentration of ACA additive in the mobile phase, control over the retention and chiral recognition of the stationary phase is demonstrated. PMID- 12725694 TI - A new gas chromatographic methodology for the estimation of the composition of binary gas mixtures. AB - The relatively new technique of reversed-flow gas chromatography (RFGC) is used to determine the diffusion coefficients of pure gases into gas mixtures (D(mix)(exp)). The pure gases are CO and CO(2), and the mixtures consist of H(2) and He in various volume percentage compositions. A linear regression analysis of D(mix)(exp) of CO and CO(2) in various mixtures of H(2) and He against the percentage composition (X(H2) or X(He)) of the mixtures at different temperatures results in an empirical equation relating D(mix)(exp) to the corresponding theoretical values of the diffusion coefficients of CO and CO(2) in the pure gases H(2) and He, as they are calculated from the Fuller-Schettler-Giddings equation. The empirical equation shows that the diffusion coefficient of an analyte gas in a gas mixture is the partial sum of its diffusion coefficients in the component gases, therefore making possible the determination of the mole fractions of the components of the mixture. The found percentage volume compositions are very close to those determined independently by routine gas chromatography, indicating that the proposed RFGC methodology could be successfully applied to the accurate determination of the volume composition of binary gas mixtures. PMID- 12725695 TI - Supercoiled circular DNA retention in nonequilibrium chromatography: viscosity and velocity dependence--behavior difference with proteins. AB - This study demonstrates that the retention behavior of various circular double stranded DNA molecules (3, 5, and 10 kb) increases over the entire flow-rate range (0.02-1.8 mL/min) at all the mobile phase viscosities (h). The transition between the two well-known nonequilibrium chromatography methods (slalom and hydrodynamic chromatography) is clearly visualized for proteins and does not appear for plasmids because of their strong compact structure. Also, the optimal conditions for F and h are determined to obtain the most efficient separation of these three plasmids in a minimum analysis time. PMID- 12725696 TI - Study on the protein binding of ketoprofen using capillary electrophoresis frontal analysis compared with liquid chromatography frontal analysis. AB - A method of capillary electrophoresis frontal analysis (CEFA) is developed for the first time to study the binding of ketoprofen to human serum albumin (HSA) and compared with high-performance liquid chromatography frontal analysis (LCFA). The separation is performed in an uncoated fused-silica capillary (60-cm x 75- micro m i.d., 50-cm effective length) with a phosphate buffer (pH 7.4, ionic strength of 0.17M) as the running buffer. The applied voltage is 13 kV and the detection is set at 254 nm. A trapezoidal peak of the unbound ketoprofen appears after HSA elution in the electropherogram. The plateau height of the peak is employed to determine the unbound concentration of ketoprofen in the HSA equilibrated sample solution. The CEFA method provides the advantages of small sample injection volume and rapidity and the disadvantage of low sensitivity compared with LCFA. CEFA is applicable to the binding parameter estimation of ketoprofen to the secondary binding site; an association constant (K(2)) of 0.24 x 10(6)M(-1) and the number for the binding site per molecule HSA of 2.54 is estimated. In contrast, LCFA measures parameters for both primary and secondary sites, which are 1.05 x 10(6)M(-1) and 0.94 for K(1) and n(1), respectively, and 0.12 x 10(6)M(-1) and 3.16 for K(2) and n(2), respectively. It is found that ketoprofen binds mainly at the primary site at a molecular ratio of ketoprofen versus HSA lower than 0.75, and the binding at the secondary site occurs at a higher ratio. PMID- 12725697 TI - Separation and determination of phospholipids in biological samples by high performance liquid chromatography. AB - An isocratic high-performance liquid chromatographic method is developed for the determination of phospholipids in biological samples using a muPorasil silica column and a mobile phase of acetonitrile-methanol-85% phosphoric acid (90:3:1, v/v/v) at a flow rate of 0.80 mL/min. The effluent is monitored by a UV detector at 203 nm. With the method reported in this paper, phosphatidylinostol, phosphatidylserine, phosphatidylethnolamine, and phosphatidylcholine in biological samples are separated and detected successfully. The method is simple, rapid, and has excellent precision. PMID- 12725698 TI - Quantitation in multianalyte overlapping peaks from capillary electrophoresis runs using artificial neural networks. AB - The potentiality of artificial neural networks for multicomponent analysis in unresolved peaks from capillary electrophoresis (CE) is evaluated. The system chosen consists of mixtures of three ebrotidine metabolites, which cannot be successfully separated by CE. Data selected for analysis consist of UV spectra taken at the maximum of the CE peak. The most dissimilar analyte, in terms of spectral differences, is accurately quantitated in any type of mixture with an overall prediction error of 5%. Because of the strong interference of the two most overlapped compounds, a preliminary procedure for spectral data filtering based on principal component analysis is performed to improve their quantitation. PMID- 12725699 TI - Sample preparation and determination of acetylcholine in corneal epithelium cells using liquid chromatography-tandem mass spectrometry. AB - A sample preparation method with subsequent liquid chromatography (LC)-mass spectrometry (MS)-MS analysis for acetylcholine in corneal epithelium is developed. The sample preparation is developed with a focus on compatibility with the LC-MS-MS system and the stability of acetylcholine because acetylcholine esterase is present in the tissue. It appears that both acetylcholine as well as the internal standard (IS) used (acetyl-beta-methylcholine) have fragments at m/z values in the tandem MS spectrum, which correspond with the m/z values of fragments of endogenous substances. Acetylcholine and (3 carboxypropyl)triethylammonium both have 146-->87 and 146-->60 transitions. Acetyl-beta-methylcholine and an unknown compound both have 160-->101 and 160- >60 transitions. This makes it necessary to use a chromatographic step, which has a baseline separation between these endogenous compounds, acetylcholine, and the IS. The analytical procedure has linearity from 1 ng/mL (30 pg/mg corneal epithelium tissue) to at least 250 ng/mL (7.55 ng/mg corneal epithelium tissue). The limits of detection and quantitation are 15 and 45 pg on column, respectively. Relative standard deviation and bias values are within the range of acceptance for all concentration levels. PMID- 12725700 TI - Investigation of host-guest stability constants of calix[n]arenes complexes with aromatic molecules by RP-HPLC method. AB - Under reversed-phase high-performance liquid chromatographic conditions [Spherisorb ODS 1 stationary phase, UV detection at 254 nm, and acetonitrile dichloromethane-acetic acid-methyl-tert-butylether (84.6/4.5/0.9/10, v/v/v/v) as the mobile phase], adding p-tert-butylcalix[8] (10(-5)-3.10(-5))-[12]arenes (10( 5)-4.10(-5) mol/L) to the mobile phase leads to decreased sorption of aromatic solutes on the surface of the sorbent because of the formation of host-guest inclusion complexes between the calixarenes and the aromatic molecules. Stability constants of the complexes (781-9338M(-1)) are determined from the relationship between the solute capacity factors and the calixarene concentration in the mobile phase. PMID- 12725701 TI - An effective absorption behavior of insulin for diabetic treatment following intranasal delivery using porous spherical calcium carbonate in monkeys and healthy human volunteers. AB - Porous spherical calcium carbonate (PS-CaCO(3)), in contrast to regular calcium carbonate (CaCO(3)), which has a cuboidal particle shape, has a characteristic spherical particle shape with a large number of porous, sliver crystals. The effect of PS-CaCO(3) as a drug carrier on intranasal insulin absorption was investigated in cynomolgus monkeys and healthy human volunteers. Each insulin formulation (powder) containing PS-CaCO(3) or regular CaCO(3) was administered intranasally. Serum insulin and glucose levels after administration were evaluated. The insulin absorption after intranasal administration with each CaCO(3) was found to be much more rapid than that after subcutaneous administration. The serum insulin level after intranasal insulin delivery (16 U per monkey) with PS-CaCO(3) showed a higher C(max) (403.5 microU/mL) and shorter T(max) (0.167 h) when compared with regular CaCO(3). The serum glucose level reduction rate after intranasal delivery using PS-CaCO(3) was faster than that of regular CaCO(3), reflecting the difference in absorption rates. Following repeated intranasal administrations for 4 weeks in monkeys, no toxicity was observed even with a maximum insulin dose level of 25 U. Furthermore, the intranasal insulin absorption rate with PS-CaCO(3) in healthy humans was also observed to be considerably faster than that with regular CaCO(3). Effects of PS CaCO(3) on a more effective absorption behavior of insulin were considered to be the result of a greater affinity between the nasal mucosa layer and PS-CaCO(3), which is closely related to its structural characteristics. Thus, intranasal insulin delivery using PS-CaCO(3) is thought to be a safe and highly available system enabling more effective insulin absorption behavior with the appearance of endogenous postprandial insulin secretion in healthy humans. We believe that our intranasal insulin delivery system enabling a rapid and short-acting pharmacological effect against postprandial hyperglycemia will be more beneficial than pulmonary insulin delivery systems in the treatment of diabetes. PMID- 12725702 TI - Glucose correlation with light scattering patterns--a novel method for non invasive glucose measurements. AB - To correlate occlusion red near-infrared spectroscopy technology with intravenous and interstitial glucose levels, occlusion red near-infrared spectroscopy and glucose levels were measured in five subjects with diabetes mellitus during a stepped hyperinsulinemic hypoglycemic clamp. Validation was achieved using a standard error grid and linear correlations. During hyperinsulinemic hypoglycemic clamp linear correlation was significant at r = 0.836, and the intercept was 43.3 mg/dL. Of the glucose measurements, 94.3% were assigned to zones A and B of the standard error grid, the zones of the greatest therapeutic relevance. Expected interference from serum triglycerides, catecholamines, and cortisol was not apparent. These results demonstrate the applicability of occlusion red near infrared spectroscopy for in vivo glucose monitoring. The technology has implications for continuous non-invasive glucose sensing. PMID- 12725703 TI - Unrecognized hypo- and hyperglycemia in well-controlled patients with type 2 diabetes mellitus: the results of continuous glucose monitoring. AB - The aim of this study was to determine the prevalence and extent of glycemic excursions (hypo- and hyperglycemic) in elderly patients with well-controlled type 2 diabetes using a Continuous Glucose Monitor System (CGMS) (Medtronic MiniMed). Elderly patients (>65 years old) with type 2 diabetes were recruited if their glycosylated hemoglobin (HbA1c) was <7.5% and if their oral hypoglycemic therapy included a sulfonylurea. Patients were asked to undergo two consecutive 72-h periods of continuous glucose monitoring at baseline and then again at 1 month (total 288 h). Patients were asked to record four self-monitored capillary blood glucose levels each day for calibration of the monitor and also to record meal times, exercise, and symptoms of hypoglycemia. The number of hyperglycemic (>144 mg/dL), hypoglycemic (<50 mg/dL), and borderline-hypoglycemic (50-65 mg/dL) events were determined (an event was defined as a glucose value that persisted for at least 15 min with or without symptoms). Twenty-five patients (21 men, four women) 73.9 +/- 4.4 years old with an HbA1c of 6.2 +/- 0.8% were each monitored for an average of 187.57 h. The mean glucose values were: fasting, 139 +/- 40 mg/dL; 2 h post-breakfast, 167 +/- 58 mg/dL; 2 h post-lunch, 157 +/- 53 mg/dL; and 2 h post-dinner, 149 +/- 49 mg/dL. Twenty patients (80%) experienced a total of 103 hypoglycemic events, and 14 of these patients experienced 54 events where the glucose levels were 144 mg/dL 2 h postprandial) were recorded after 57% of all meals (breakfast 60%, lunch 57.5%, dinner 55.2%). The CGMS was generally well tolerated, but 52% of patients could not be studied for the full 12 days of monitoring. Thus hypoglycemia and excessive postprandial glycemic excursions are common in well-controlled patients with type 2 diabetes treated with a sulfonylurea with or without metformin. The CGMS is a useful research and clinical tool to assess glycemia in patients with type 2 diabetes but is not tolerated by all subjects. PMID- 12725704 TI - Determination of plasma glucose during rapid glucose excursions with a subcutaneous glucose sensor. AB - Continuous glucose monitoring has the potential to improve glucose management and reduce the risk of hypoglycemia in individuals with diabetes. Accurate sensors may also allow the development of a closed-loop insulin delivery system. The purpose of this work was to determine the delay time associated with a subcutaneous glucose sensor during rapidly changing glucose excursions. Subcutaneous glucose sensors (Medtronic MiniMed, Inc., Northridge, CA) were inserted in five healthy men. After a 2-h stabilization period, a 3-h hyperglycemic (approximately 11 mM) clamp was performed followed by a 90-min period in which plasma glucose was allowed to decline to as low as 2.8 mM. Sensors were calibrated using two points (basal and hyperglycemia), and the calibrated sensor glucose measurements were compared with those from a reference analyzer (Beckman Instruments, Fullerton, CA). Response time was estimated from a first-order kinetic model. Plasma glucose levels, determined with the subcutaneous sensor, were highly correlated with those obtained with the reference glucose analyzer (r(2) = 0.91, p < 0.001; mean absolute difference of approximately 8%). The half-time for the sensor response was estimated to be 4.0 +/- 1.0 min. The subcutaneous glucose sensor has the potential to facilitate the detection of hypoglycemia and improve overall glycemic control when used in a real-time monitor. The rapid response should be sufficient to allow a fully automated closed-loop insulin delivery system to be developed based on the subcutaneous sensing site. PMID- 12725705 TI - Addition of rosiglitazone to existing sulfonylurea treatment in chinese patients with type 2 diabetes and exposure to hepatitis B or C. AB - The effects of adding rosiglitazone to existing sulfonylurea (SU) treatment have not previously been studied in Chinese patients with type 2 diabetes and no known pre-existing hepatic impairment. Patients were randomized to receive rosiglitazone 2 mg twice daily (R4 + SU) or 4 mg twice daily (R8 + SU) or placebo (SU + P) for 24 weeks in addition to existing SU treatment. Most patients were taking concomitant glibenclamide (34%) or gliclazide (25%). Changes in glycosylated hemoglobin (HbA(1c)), fasting plasma glucose (FPG), and plasma insulin concentrations were measured. Of the 530 patients enrolled (45% male, mean age 59 years), 105 were in the SU + P group, 215 in the R4 + SU group, and 210 in the R8 + SU group. The mean baseline HbA(1c) was 9.8%, and FPG was 183.8 mg/dL. Compared with placebo, addition of rosiglitazone (2 or 4 mg twice daily) produced significant decreases in mean HbA(1c) (1.04% and 1.44%, respectively; p < 0.0001) and FPG (21.6 and 36.0 mg/dL, respectively; p < 0.0001). There were statistically significant (p < 0.0001) reductions from baseline in insulin concentration of 23.3 and 30.4 pmol/L in the R4 + SU and R8 + SU groups, respectively. Despite the high prevalence of seropositivity for hepatitis B and/or C at baseline (56%), there was no evidence of hepatotoxicity. No clinically significant changes in routine hematology, biochemistry, or electrocardiogram were observed. The addition of rosiglitazone to SU produced clinically significant improvements in glycemic control in Chinese patients with type 2 diabetes. Rosiglitazone plus SU was well tolerated irrespective of hepatitis B and C serological status. PMID- 12725706 TI - Glitazones: good for glycemia. Good for the liver? PMID- 12725707 TI - Inadequacy of in-school support for diabetic children. AB - The level of blood glucose control needed to minimize complications in children with diabetes requires frequent blood sugar monitoring and appropriate responses to the information obtained. It is our impression that optimal support for good control is not available in all of the schools our patients attend. The objective of this study was to identify and quantify barriers to good control of diabetes in the school setting, and then use this information to target interventions to improve in-school support for children with diabetes. Two questionnaires were designed based on recommendations of the American Diabetes Association for appropriate in-school support for children with diabetes. Parental perception of in-school resources was addressed in one questionnaire. Forty-seven parents of children with diabetes in our clinic were surveyed. The second questionnaire was mailed to 222 randomly selected schools in our area inquiring about the in-school support available to children with diabetes and the types of educational materials that would be useful for school personnel. Thirty percent of the parents of children with diabetes indicated that the in-school support of their child was insufficient. Sixty-five of the 222 schools surveyed responded. The responses were variable and demonstrated inconsistency and, in some cases, inadequacy of support. A major deficiency noted in 13% of schools was lack of on site personnel trained in diabetes management skills. From the schools' perspective, however, 50% of schools reported lack of parental communication. The care available for the child with diabetes is highly variable among schools. Targeted educational materials for both school personnel and parents would be useful to improve support for these children. PMID- 12725708 TI - Effects of rapid-acting insulin analogs on overall glycemic control in type 1 and type 2 diabetes mellitus. AB - The altered pharmacokinetics of new rapid-acting insulin analogs make them very effective in controlling blood glucose peaks after meals. However, simple replacement of regular human insulin by rapid-acting analogs may not be sufficiently effective for overall daily glycemic control. Both basal and bolus insulins need to be addressed with overall changes in insulin regimens for long term reductions of glycated hemoglobin (HbA(1c)) levels. Clinical studies in patients with type 1 diabetes have shown that better control of postprandial peaks together with optimal use of basal insulin, in multiple injection regimens or with continuous subcutaneous insulin infusion, resulted in improved HbA(1c) levels of 0.3-0.4% in comparison with regular human insulin. In patients with type 2 diabetes the combination therapy of insulin lispro with an oral agent improved metabolic control in the range of 1.5-2.5%. Such decreases in HbA(1c) should lead to a reduction in risk of complications with both type 1 and type 2 diabetes. The better overall metabolic control with rapid-acting insulin analogs was not accompanied by any increased risk for hypoglycemia. PMID- 12725709 TI - Therapeutic potential of peroxisome proliferator-activated receptor agonists for neurological disease. AB - Activation of peroxisome proliferator-activated receptors (PPARs) mediates the insulin-sensitizing effects of thiazolidinediones used for treatment of type 2 diabetes, owing to changes in the transcription and expression of genes influencing carbohydrate and lipid metabolism. However, PPAR activation can have additional effects upon cellular physiology, including anti-proliferative and anti-inflammatory. These effects are observed in many cell types, including brain glial cells and blood lymphocytes, cells whose activation contributes to the initiation and progression of damage occurring in neurological diseases such as Alzheimer's disease (AD) and multiple sclerosis (MS). In view of the need for development of additional therapeutic options, several recent studies have tested the possibility that PPAR agonists would be neuroprotective in these diseases. This paper will summarize data from cell culture experiments and from studies in animal models, demonstrating that PPARgamma agonists can exert neuroprotective effects, thereby providing the basis for the design of clinical trials to test the safety and efficacy of thiazolidinediones in neuroinflammatory conditions such as AD and MS. PMID- 12725710 TI - Usage of the www.2aida.org AIDA diabetes software Website: a pilot study. AB - AIDA is a diabetes-computing program freely available from www.2aida.org on the Web. The software is intended to serve as an educational support tool, and can be used by anyone who has an interest in diabetes, whether they be patients, relatives, health-care professionals, or students. In previous "Diabetes Information Technology & WebWatch" columns various indicators of usage of the AIDA program have been reviewed, and various comments from users of the software have been documented. One aspect of AIDA, though, that has been of considerable interest has been to investigate its Web-based distribution as a wider paradigm for more general medically related usage of the Internet. In this respect we have been keen to understand in general terms: (1) why people are turning to the Web for health-care/diabetes information; (2) more specifically, what sort of people are making use of the AIDA software; and (3) what benefits they feel might accrue from using the program. To answer these types of questions we have been conducting a series of audits/surveys via the AIDA Website, and via the software program itself, to learn as much as possible about who the AIDA end users really are. The rationale for this work is that, in this way, it should be possible to improve the program as well as tailor future versions of the software to the interests and needs of its users. However, a recurring observation is that data collection is easiest if it is as unobtrusive and innocuous as possible. One aspect of learning as much as possible about diabetes Website visitors and users may be to apply techniques that do not necessitate any visitor or user interaction. There are various programs that can monitor what pages visitors are viewing at a site. As these programs do not require visitors to do anything special, over time some interesting insights into Website usage may be obtained. For the current study we have reviewed anonymous logstats data, which are automatically collected at many Websites, to try and establish a baseline level of usage for the AIDA site. For the initial pilot study the analysis was performed from October 1, 2000 to November 1, 2001. The study has yielded an interesting insight into how the AIDA Website is being used. The results also confirm those of previous audits based on different self-reported methodologies, confirming, amongst other things, what countries people are visiting from and what operating systems/computers they are using. These analyses have been informative and useful. Given this, it is proposed to repeat the current pilot survey approach on a routine basis, in the future, as a way of monitoring on going usage of the AIDA Website. PMID- 12725713 TI - Selecting items for a food behavior checklist for a limited-resource audience. AB - OBJECTIVE: To report 6 psychometric properties of food behavior checklist (FBC) items and then to use these properties to systematically reduce the number of items on this evaluation tool. DESIGN: Random assignment to the intervention and control groups. SETTING: Low-income communities. PARTICIPANTS: Women (N = 132) from limited-resource families. MAIN OUTCOME MEASURES: Reliability, internal consistency, baseline differences by ethnicity, sensitivity to change, and criterion and convergent validity of subscales. RESULTS: The fruit and vegetable subscale showed a significant correlation with serum carotenoid values (r =.44, P <.001), indicating acceptable criterion validity. Milk, fat/cholesterol, diet quality, food security, and fruit/vegetable subscales showed significant correlations with dietary variables. Nineteen items have acceptable reliability. Twenty items showed no baseline differences by ethnic group. Eleven of the 15 items expected to show change following the intervention demonstrated sensitivity to change. CONCLUSIONS AND IMPLICATIONS: This brief food behavior checklist (16 items) is easy to administer to a client group, has an elementary reading level (fourth grade), and has a low respondent burden in addition to meeting requirements for validity, reliability, and sensitivity to change. This study establishes a process that can be used by other researchers to develop and further refine instruments for use in community health promotion interventions. PMID- 12725714 TI - Reliability and validity of nutrition knowledge, social-psychological factors, and food label use scales from the 1995 Diet and Health Knowledge Survey. AB - OBJECTIVE: To test the reliability and validity of scales on nutrition knowledge, social-psychological factors, and use of food labels developed from the 1995 Diet and Health Knowledge Survey (DHKS) questions. DESIGN: The 1995 DHKS questions within a section were pooled together as a scale and their reliability and validity were examined. PARTICIPANTS: US adults (> or =20 years) in the 1995 DHKS who responded to questions selected for this study (n = 1196). VARIABLES: Nutrition knowledge about the diet-disease relationship and nutrient content of products, perceived barriers and benefits of food labels, perceived ease of understanding food labels, food label use, and importance of healthful eating. ANALYSIS: Scales validity, Cronbach alpha, item total correlation, alpha if the item was deleted, and discriminant, convergence, and correspondence validity. RESULTS: Scales on perceived ease of understanding the food label, benefits of using food labels, food label use, and importance of healthful eating were reliable (Cronbach alpha =.78,.82,.91, and.82, respectively) and valid. CONCLUSION AND IMPLICATIONS: Accurate findings and interpretation of survey data depend on the use of reliable and valid instruments. This study identified the scales in the DHKS that can substantiate the conclusion on which effective nutrition education strategies should be established. PMID- 12725715 TI - Validation of a fruit, juice, and vegetable availability questionnaire. AB - OBJECTIVE: To assess the validity of a self-reported fruit, juice, and vegetable home availability measure. DESIGN: Cross-sectional. SETTING: In-home interviews. PARTICIPANTS: Forty-eight parents of fourth to sixth grade children. MAIN OUTCOME MEASURES: Scales assessing self-reported home availability of fruit, 100% fruit juice, and vegetables over the past 7 days compared with an observed home inventory conducted on the interview day. ANALYSES: Cohen's kappa of agreement, specificity and sensitivity, and correlations, with significance level <.05. RESULTS: There was substantial agreement between self-reported and observed home inventories of many fruit, 100% fruit juice, and vegetables among participating families in this study. CONCLUSIONS AND IMPLICATIONS: Self-reported shelf inventories can be an effective tool for use in assessing the availability of fruit, juice, and vegetables in the home. PMID- 12725716 TI - Desire to eat high- and low-fat foods following a low-fat dietary intervention. AB - OBJECTIVE: This study examined changes in desires to eat high-fat and low-fat foods across an obesity treatment program. The hypotheses under examination were (1) preferences for low-fat foods would increase across time and (2) preferences for high-fat foods would decrease across time. DESIGN: Single-group, prospective examination of desires to eat 48 foods, categorized according to fat content, before and after the 16-week treatment program. SETTING: University clinic, Memphis, Tennessee. PARTICIPANTS: 118 obese (mean weight = 194.4 lbs) women (mean age = 45.24 years) participating in an obesity treatment program. INTERVENTION: A 16-week cognitive-behavioral program for obesity. VARIABLES MEASURED: Desires to eat 48 foods varying in fat content and whether or not participants actually ate these foods. ANALYSIS: Analysis of variance, multiple regression, and paired t tests. RESULTS: The results indicate that during the program, preferences for low fat foods increased, whereas preferences for high-fat foods decreased. These changes mirrored the changes in consumption of both low-fat and high-fat foods. CONCLUSIONS AND IMPLICATIONS: Within a behavioral economic perspective, the reinforcement value of low-fat foods may increase following a low-fat dietary intervention, whereas the reinforcing properties of high-fat foods may decline. This is desirable as low-fat foods hold many advantages over high-fat foods in terms of weight maintenance. PMID- 12725717 TI - Teaching elementary school children about healthy Native American foods. PMID- 12725718 TI - Cooking with class: participation soars with hands-on learning and take-aways. PMID- 12725720 TI - Relevancy of drug loading to liposomal formulation therapeutic efficacy. PMID- 12725721 TI - Animal experiments-an essential component for the development of liposomal anticancer agents. PMID- 12725722 TI - Emerging role of liposomal drug carrier systems in cancer chemotherapy. PMID- 12725723 TI - Dendritic cells capture and efficiently present antigen encapsulated in liposomes to T cells in vivo. PMID- 12725724 TI - Anti-neovascular therapy by use of liposomes targeted to angiogenic vessels. PMID- 12725726 TI - Liposomes: from the bench to the bed. PMID- 12725725 TI - Magnetofection: enhancing and targeting gene delivery with superparamagnetic nanoparticles and magnetic fields. PMID- 12725727 TI - PAR proteins regulate microtubule dynamics at the cell cortex in C. elegans. AB - BACKGROUND: The PAR proteins are known to be localized asymmetrically in polarized C. elegans, Drosophila, and human cells and to participate in several cellular processes, including asymmetric cell division and spindle orientation. Although astral microtubules are known to play roles in these processes, their behavior during these events remains poorly understood. RESULTS: We have developed a method that makes it possible to examine the residence time of individual astral microtubules at the cell cortex of developing embryos. Using this method, we found that microtubules are more dynamic at the posterior cortex of the C. elegans embryo compared to the anterior cortex during spindle displacement. We further observed that this asymmetry depends on the PAR-3 protein and heterotrimeric G protein signaling, and that the PAR-2 protein affects microtubule dynamics by restricting PAR-3 activity to the anterior of the embryo. CONCLUSIONS: These results indicate that PAR proteins function to regulate microtubule dynamics at the cortex during microtubule-dependent cellular processes. PMID- 12725728 TI - The UCS domain protein She4p binds to myosin motor domains and is essential for class I and class V myosin function. AB - BACKGROUND: Myosins are motor proteins involved in processes like cell motility, vesicle transport, or cytokinesis. In a variety of organisms, a novel group of proteins forming the UCS (UNC-45/CRO1/SHE4) domain-containing family are essential for proper myosin function. The Saccharomyces cerevisae UCS domain protein She4p is involved in two myosin-requiring events, endocytosis and mRNA localization. RESULTS: In contrast to UCS domain proteins from other organisms that interact with class II myosins, we demonstrate that She4p associates with yeast class I and class V myosins. She4p binds to motor domains of class V myosin Myo4p and class I myosin Myo5p, and this binding depends on She4p's UCS domain. In vivo, She4p is essential for the function and localization of Myo3p, Myo4p, and Myo5p (but not of Myo2p) and for colocalization of class I myosins with cortical actin patches. In vitro, She4p stimulates binding of Myo5p to filamentous actin. Wild-type She4p, but not a mutant lacking the UCS domain, accumulates in a cap-like structure at the bud tip. This localization requires Myo2p and actin, suggesting a Myo2-dependent mechanism by which She4p is targeted to the bud cap. Localization of She4p is essential for proper positioning and myosin-actin association of cortical Myo5p. CONCLUSIONS: Our results suggest that She4p is a novel myosin motor domain binding protein and operates as a localized regulator of myosin function of class I and likely class V myosins. PMID- 12725729 TI - cGMP-dependent protein kinase II modulates mPer1 and mPer2 gene induction and influences phase shifts of the circadian clock. AB - BACKGROUND: In mammals, the master circadian clock that drives many biochemical, physiological, and behavioral rhythms is located in the suprachiasmatic nuclei (SCN) of the hypothalamus. Generation and maintenance of circadian rhythmicity rely on complex interlocked transcriptional/translational feedback loops involving a set of clock genes. Among the molecular components driving the mammalian circadian clock are the Period 1 and 2 (mPer1 and mPer2) genes. Because the periodicity of the clock is not exactly 24 hr, it has to be adjusted periodically. The major stimulus for adjustment (resetting) of the clock is nocturnal light. It evokes activation of signaling pathways in the SCN that ultimately lead to expression of mPer1 and mPer2 genes conveying adjustment of the clock. RESULTS: We show that mice deficient in cGMP-dependent protein kinase II (cGKII, also known as PKGII), despite regular retinal function, are defective in resetting the circadian clock, as assessed by changes in the onset of wheel running activity after a light pulse. At the molecular level, light induction of mPer2 in the SCN is strongly reduced in the early period of the night, whereas mPer1 induction is elevated in cGKII-deficient mice. Additionally, we show that light induction of cfos and light-dependent phosphorylation of CREB at serine 133 are not affected in these animals. CONCLUSIONS: cGKII plays a role in the clock resetting mechanism. In particular, the ability to delay clock phase is affected in cGKII-deficient mice. It seems that the signaling pathway involving cGKII influences in an opposite manner the light-induced induction of mPer1 and mPer2 genes and thereby influences the direction of a phase shift of the circadian clock. PMID- 12725730 TI - Mammalian Lgl forms a protein complex with PAR-6 and aPKC independently of PAR-3 to regulate epithelial cell polarity. AB - BACKGROUND: Epithelial cells have apicobasal polarity and an asymmetric junctional complex that provides the bases for development and tissue maintenance. In both vertebrates and invertebrates, the evolutionarily conserved protein complex, PAR-6/aPKC/PAR-3, localizes to the subapical region and plays critical roles in the establishment of a junctional complex and cell polarity. In Drosophila, another set of proteins called tumor suppressors, such as Lgl, which localize separately to the basolateral membrane domain but genetically interact with the subapical proteins, also contribute to the establishment of cell polarity. However, how physically separated proteins interact remains to be clarified. RESULTS: We show that mammalian Lgl competes for PAR-3 in forming an independent complex with PAR-6/aPKC. During cell polarization, mLgl initially colocalizes with PAR-6/aPKC at the cell-cell contact region and is phosphorylated by aPKC, followed by segregation from apical PAR-6/aPKC to the basolateral membrane after cells are polarized. Overexpression studies establish that increased amounts of the mLgl/PAR-6/aPKC complex suppress the formation of epithelial junctions; this contrasts with the previous observation that the complex containing PAR-3 promotes it. CONCLUSIONS: These results indicate that PAR-6/aPKC selectively interacts with either mLgl or PAR-3 under the control of aPKC activity to regulate epithelial cell polarity. PMID- 12725731 TI - Evidence for preferential mismatch repair of lagging strand DNA replication errors in yeast. AB - Duplex DNA is replicated in the 5'-3' direction by coordinated copying of leading and lagging strand templates with somewhat different proteins and mechanics, providing the potential for differences in the fidelity of replication of the two strands. We previously showed that in Saccharomyces cerevisiae, active replication origins establish a strand bias in the rate of base substitutions resulting from replication of unrepaired 8-oxo-guanine (GO) in DNA. Lower mutagenesis was associated with replicating lagging strand templates. Here, we test the hypothesis that this bias is due to more efficient repair of lagging stand mismatches by measuring mutation rates in ogg1 strains with a reporter allele in two orientations at loci on opposite sides of a replication origin on chromosome III. We compare a MMR-proficient strain to strains deleted for the MMR genes MSH2, MSH6, MLH1, or EXOI. Loss of MMR reduces the strand bias by preferentially increasing mutagenesis for lagging strand replication. We conclude that GO-A mismatches generated during lagging strand replication are more efficiently repaired. This is consistent with the hypothesis that 5' ends of Okazaki fragments and PCNA, present at high density during lagging strand replication, are used as strand discrimination signals for mismatch repair in vivo. PMID- 12725732 TI - Analogous mechanisms compensate for neural delays in the sensory and the motor pathways: evidence from motor flash-lag. AB - Motor behaviors require animals to coordinate neural activity across different areas within their motor system. In particular, the significant processing delays within the motor system must somehow be compensated for. Internal models of the motor system, in particular the forward model, have emerged as important potential mechanisms for compensation. For motor responses directed at moving visual objects, there is, additionally, a problem of delays within the sensory pathways carrying crucial position information. The visual phenomenon known as the flash-lag effect has led to a motion-extrapolation model for compensation of sensory delays. In the flash-lag effect, observers see a flashed item colocalized with a moving item as lagging behind the moving item. Here, we explore the possibility that the internal forward model and the motion-extrapolation model are analogous mechanisms compensating for neural delays in the motor and the visual system, respectively. In total darkness, observers moved their right hand gripping a rod while a visual flash was presented at various positions in relation to the rod. When the flash was aligned with the rod, observers perceived it in a position lagging behind the instantaneous felt position of the invisible rod. These results suggest that compensation of neural delays for time-varying motor behavior parallels compensation of delays for time-varying visual stimulation. PMID- 12725733 TI - Experimental evidence for the evolution of numerous, tiny sperm via sperm competition. AB - Sperm competition, when sperm from different males compete to fertilize a female's ova, is a widespread and fundamental force in the evolution of animal reproduction. The earliest prediction of sperm competition theory was that sperm competition selected for the evolution of numerous, tiny sperm, and that this force maintained anisogamy. Here, we empirically test this prediction directly by using selective breeding to generate controlled and independent variance in sperm size and number traits in the cricket Gryllus bimaculatus. We find that sperm size and number are male specific and vary independently and significantly. We can therefore noninvasively screen individuals and then run sperm competition experiments between males that differ specifically in sperm size and number traits. Paternity success across 77 two-male sperm competitions (each running over 30-day oviposition periods) shows that males producing both relatively small sperm and relatively numerous sperm win competitions for fertilization. Decreased sperm size and increased sperm number both independently predicted sperm precedence. Our findings provide direct experimental support for the theory that sperm competition selects for maximal numbers of miniaturized sperm. However, our study does not explain why G. bimaculatus sperm length persists naturally at approximately 1 mm; we discuss possibilities for this sperm size maintenance. PMID- 12725734 TI - A novel C-terminal domain of drosophila PERIOD inhibits dCLOCK:CYCLE-mediated transcription. AB - The essence of the Drosophila circadian clock involves an autoregulatory feedback loop in which PERIOD (PER) and TIMELESS (TIM) inhibit their own transcription by association with the transcriptional activators dCLOCK (dCLK) and CYCLE (CYC). Because PER, dCLK, and CYC each contain a PAS domain, it has been assumed that these interaction domains are important for negative feedback. However, a critical role for PAS-PAS interactions in Drosophila clock function has not been shown. Nuclear transport of PER is also believed to be an essential regulatory step for negative feedback, but this has not been directly tested, and the relevant nuclear localization sequence (NLS) has not been functionally mapped. We evaluated these critical aspects of PER-mediated transcriptional inhibition in Drosophila Schneider 2 (S2) cells. We mapped the dCLK:CYC inhibition domain (CCID) of PER and discovered that it lies in the C terminus, downstream of the PAS domain. Using deletion mutants and site-directed mutagenesis, we identified a novel NLS in the CCID of PER that is a potent regulator of PER's nuclear transport in S2 cells. We further found that nuclear transport, primarily through this novel NLS, is essential for the inhibitory activity of PER. The data indicate that nuclear PER inhibits dCLK:CYC-mediated transcription through a novel domain that additionally contains a potent NLS. PMID- 12725735 TI - A simple reason for a big difference: wolves do not look back at humans, but dogs do. AB - The present investigations were undertaken to compare interspecific communicative abilities of dogs and wolves, which were socialized to humans at comparable levels. The first study demonstrated that socialized wolves were able to locate the place of hidden food indicated by the touching and, to some extent, pointing cues provided by the familiar human experimenter, but their performance remained inferior to that of dogs. In the second study, we have found that, after undergoing training to solve a simple manipulation task, dogs that are faced with an insoluble version of the same problem look/gaze at the human, while socialized wolves do not. Based on these observations, we suggest that the key difference between dog and wolf behavior is the dogs' ability to look at the human's face. Since looking behavior has an important function in initializing and maintaining communicative interaction in human communication systems, we suppose that by positive feedback processes (both evolutionary and ontogenetically) the readiness of dogs to look at the human face has lead to complex forms of dog-human communication that cannot be achieved in wolves even after extended socialization. PMID- 12725736 TI - Direction repulsion goes global. AB - When viewing two superimposed, translating sets of dots moving in different directions, one overestimates direction difference. This phenomenon of direction repulsion is thought to be driven by inhibitory interactions between directionally tuned motion detectors. However, there is disagreement on where this occurs-at early stages of motion processing, when local motions are extracted; or at the later, global motion-processing stage following "pooling" of these local measures. These two stages of motion processing have been identified as occurring in area V1 and the human homolog of macaque MT/V5, respectively. We designed experiments in which local and global predictions of repulsion are pitted against one another. Our stimuli contained a target set of dots, moving at a uniform speed, superimposed on a "mixed-speed" distractor set. Because the perceived speed of a mixed-speed stimulus is equal to the dots' average speed, a global-processing account of direction repulsion predicts that repulsion magnitude induced by a mixed-speed distractor will be indistinguishable from that induced by a single-speed distractor moving at the same mean speed. This is exactly what we found. These results provide compelling evidence that global motion interactions play a major role in driving direction repulsion. PMID- 12725737 TI - Assembly of a mediator/TFIID/TFIIA complex bypasses the need for an activator. AB - Transcription in eukaryotic cells requires the remodeling of chromatin and the assembly of functional preinitiation complexes (PICs), which contain the general transcription factors (GTFs), RNA polymerase II (Pol II), and coactivators. Genetic and biochemical studies have implicated the multisubunit Mediator coactivator complex (Med) as a critical component of the PIC, a direct target of activators, and a checkpoint for regulated gene expression during differentiation, development (reviewed in ), signaling, and oncogenesis. In this report, we show that a complex containing the activator GAL4-VP16, Med, and TFIID/TFIIA (DA) recruits pol II and the remaining GTFs to a model promoter in vitro. A preassembled DAMed complex bypasses the requirement for an activator. We also demonstrate that coordinated assembly of DAMed is essential to establishing a functional PIC. We conclude that the DAMed complex generates a platform that supports activated levels of PIC assembly and transcription. PMID- 12725738 TI - Lethal giant larvae acts together with numb in notch inhibition and cell fate specification in the Drosophila adult sensory organ precursor lineage. AB - The tumor suppressor genes lethal giant larvae (lgl) and discs large (dlg) act together to maintain the apical basal polarity of epithelial cells in the Drosophila embryo. Neuroblasts that delaminate from the embryonic epithelium require lgl to promote formation of a basal Numb and Prospero crescent, which will be asymmetrically segregated to the basal daughter cell upon division to specify cell fate. Sensory organ precursors (SOPs) also segregate Numb asymmetrically at cell division. Numb functions to inhibit Notch signaling and to specify the fates of progenies of the SOP that constitute the cellular components of the adult sensory organ. We report here that, in contrast to the embryonic neuroblast, lgl is not required for asymmetric localization of Numb in the dividing SOP. Nevertheless, mosaic analysis reveals that lgl is required for cell fate specification within the SOP lineage; SOPs lacking Lgl fail to specify internal neurons and glia. Epistasis studies suggest that Lgl acts to inhibit Notch signaling by functioning downstream or in parallel with Numb. These findings uncover a previously unknown function of Lgl in the inhibition of Notch and reveal different modes of action by which Lgl can influence cell fate in the neuroblast and SOP lineages. PMID- 12725739 TI - Negative feedback regulation of Dicer-Like1 in Arabidopsis by microRNA-guided mRNA degradation. AB - Formation of microRNA (miRNA) requires an RNaseIII domain-containing protein, termed DICER-1 in animals and DICER-LIKE1 (DCL1) in plants, to catalyze processing of an RNA precursor with a fold-back structure. Loss-of-function dcl1 mutants of Arabidopsis have low levels of miRNA and exhibit a range of developmental phenotypes in vegetative, reproductive, and embryonic tissues. In this paper, we show that DCL1 mRNA occurs in multiple forms, including truncated molecules that result from aberrant pre-mRNA processing. Both full-length and truncated forms accumulated to relatively low levels in plants containing a functional DCL1 gene. However, in dcl1 mutant plants, dcl1 RNA forms accumulated to levels several-fold higher than those in DCL1 plants. Elevated levels of DCL1 RNAs were also detected in miRNA-defective hen1 mutant plants and in plants expressing a virus-encoded suppressor of RNA silencing (P1/HC-Pro), which inhibits miRNA-guided degradation of target mRNAs. A miRNA (miR162) target sequence was predicted near the middle of DCL1 mRNA, and a DCL1-derived RNA with the properties of a miR162-guided cleavage product was identified and mapped. These results indicate that DCL1 mRNA is subject to negative feedback regulation through the activity of a miRNA. PMID- 12725740 TI - The Drosophila microRNA Mir-14 suppresses cell death and is required for normal fat metabolism. AB - MicroRNAs (miRNAs) are small regulatory RNAs that are between 21 and 25 nucleotides in length and repress gene function through interactions with target mRNAs. The genomes of metazoans encode on the order of several hundred miRNAs, but the processes they regulate have been defined for only two in C. elegans. We searched for new inhibitors of apoptotic cell death by testing existing collections of P element insertion lines for their ability to enhance a small-eye phenotype associated with eye-specific expression of the Drosophila cell death activator Reaper. Here we report the identification of the Drosophila miRNA mir 14 as a cell death suppressor. Loss of mir-14 enhances Reaper-dependent cell death, whereas ectopic expression suppresses cell death induced by multiple stimuli. Animals lacking mir-14 are viable. However, they are stress sensitive and have a reduced lifespan. Mir-14 mutants have elevated levels of the apoptotic effector caspase Drice, suggesting one potential site of action. Mir-14 also regulates fat metabolism. Deletion of mir-14 results in animals with increased levels of triacylglycerol and diacylglycerol, whereas increases in mir-14 copy number have the converse effect. We discuss possible relationships between these phenotypes. PMID- 12725742 TI - US model pushed for UK managers. PMID- 12725743 TI - Fiona Watt. PMID- 12725741 TI - Editors sound the SARS alarm bells. PMID- 12725744 TI - A Grand Prix performance. PMID- 12725746 TI - Fibroblast growth factors. PMID- 12725747 TI - Dispatch. Animal communication: what makes a dog able to understand its master? AB - The dog has a special relationship with humans, going beyond that of other domestic animals. Recent evidence suggests this comes from domestication rather than wolf behaviour, perhaps involving something as simple as a change in natural looking behaviour. PMID- 12725748 TI - Dispatch. Intercellular signaling: an elusive player steps forth. AB - Plasmodesmata play a central role in cell-to-cell communication in plants, allowing transport of regulatory proteins and mRNAs. A recent study has identified a specific protein that regulates the intercellular transport of macromolecules in plants, known as non-cell autonomous pathway protein 1. PMID- 12725750 TI - Dispatch. Dictyostelium chemotaxis: fascism through the back door? AB - Aggregating Dictyostelium cells secrete cyclic AMP to attract their neighbours by chemotaxis. It has now been shown that adenylyl cyclase is enriched in the rear of cells, and this localisation is required for normal aggregation. PMID- 12725749 TI - Dispatch. Centrosome biology: a SAS-sy centriole in the cell cycle. AB - A novel protein in Caenorhabditis elegans, SAS-4, is a component of centrioles and is required for centriole duplication. Depletion of SAS-4 results in stunted centrioles and a smaller centrosome, suggesting a link to organelle size control. PMID- 12725751 TI - Dispatch. Sperm biology: size indeed matters. AB - Sperm length is highly variable within and across species, but relatively little attention has been paid to this variation. Two recent studies employing laboratory selection experiments have provided novel insights into the evolution of sperm size. PMID- 12725752 TI - Dispatch. Chromosome position: now, where was I? AB - Is the nuclear organisation of chromosomes inherited through mitosis, when the nuclear membrane is broken down, and is it propagated to the nuclei of daughter cells? Two recent studies address this question using similar live cell imaging techniques, but reach different conclusions. PMID- 12725753 TI - Dispatch. GTPase signalling: new functions for Diaphanous-related formins. AB - The Diaphanous-related formins are effectors for Rho GTPases. Two recent reports have unveiled previously unappreciated activities of Drf3 (mDia2), eliciting cytoskeletal rearrangements at the behest of Cdc42, and of DRF2 (hDia2C), regulating endosome dynamics for RhoD. PMID- 12725754 TI - Dispatch. Cell division: the place and time of cytokinesis. AB - Recent studies have provided evidence that, during cytokinesis, activation of the Pbl-Rho1 pathway by a protein complex located at the spindle midzone, and inhibition of this pathway by two mitotic cyclins, may be major contributing factors controlling the place and timing of the cleavage furrow. PMID- 12725755 TI - Dispatch. Gibberellin signaling: GRASs growing roots dispatch. AB - Gibberellins are important endogenous plant growth regulators. A new study has revealed an unexpected role for gibberellin-regulated transcription factors in promoting root growth; this depends on signaling from another plant hormone, auxin, suggesting that auxin promotes root growth by modulating the cellular response to gibberellins. PMID- 12725756 TI - Plant meristems: a merry-go-round of signals. AB - Recent studies have provided significant new insights into the gene actions that specify and maintain stem cells in plant shoots and roots. New layers of genetic control and potential signalling pathways and effector mechanisms have emerged from these new studies and will be reviewed here. These new findings refine the current model in which stem cells in plant meristems are regulated by negative feedback loops and uncover a fundamental mechanism for stem cell maintenance that might be common to shoots and roots. PMID- 12725758 TI - Comparison of fMRI activation at 3 and 1.5 T during perceptual, cognitive, and affective processing. AB - Previous studies comparing fMRI data acquired at 1.5 T and higher field strengths have focused on examining signal increases in the visual and motor cortices. No information is, however, available on the relative gain, or the comparability of data, obtained at higher field strengths for other brain regions such as the prefrontal and other association cortices. In the present study, we investigated fMRI activation at 1.5 and 3 T during visual perception, visuospatial working memory, and affect-processing tasks. A 23% increase in striate and extrastriate activation volume was observed at 3 T compared with that for 1.5 T during the visual perception task. During the working memory task significant increases in activation volume were observed in frontal and parietal association cortices as well as subcortical structures, including the caudate, globus pallidus, putamen, and thalamus. Increases in working memory-related activation volume of 82, 73, 83, and 36% were observed in the left frontal, right frontal, left parietal, and right parietal lobes, respectively, for 3 T compared with 1.5 T. These increases were characterized by increased activation at 3 T in several prefrontal and parietal cortex regions that showed activation at 1.5 T. More importantly, at 3 T, activation was detected in several regions, such as the ventral aspects of the inferior frontal gyrus, orbitofrontal gyrus, and lingual gyrus, which did not show significant activation at 1.5 T. No difference in height or extent of activation was detected between the two scanners in the amygdala during affect processing. Signal dropout in the amygdala from susceptibility artifact was greater at 3 T, with a 12% dropout at 3 T compared with a 9% dropout at 1.5 T. The spatial smoothness of T2* images was greater at 3 T by less than 1 mm, suggesting that the greater extent of activation at 3 T beyond these spatial scales was not due primarily to increased intrinsic spatial correlations at 3 T. Rather, the increase in percentage of voxels activated reflects increased sensitivity for detection of brain activation at higher field strength. In summary, our findings suggest that functional imaging of prefrontal and other association cortices can benefit significantly from higher magnetic field strength. PMID- 12725759 TI - Feeling-of-knowing in episodic memory: an event-related fMRI study. AB - An individual may fail to recall an item from memory but still feel that it would be recognized on a later test, a retrieval state termed the "feeling-of-knowing" (FOK). In this study we used event-related fMRI and the FOK to examine both encoding- and retrieval-related factors that are associated with different levels of recall performance: successful retrieval of a previously studied item, retrieval failure accompanied by the FOK, and retrieval failure without any FOK. The results revealed one predominant pattern of retrieval-related activation: an intermediate level of activation for FOK-less than that associated with successful recall and greater than that associated with unsuccessful recall (frontal and left parietal cortices). Two further patterns were also observed: greater activation for both successful recall and FOK than for unsuccessful recall (left midlateral prefrontal cortex) and greater activation for successful recall than for both FOK and unsuccessful recall (left MTL). Analysis of encoding trials conditional upon subsequent retrieval success revealed a pattern of activation that appeared to predict subsequent recall, but which further analysis indicated to be a better predictor of subsequent recognition. These results provide evidence that the phenomenology of graded recall is represented neurally in frontal and parietal cortices, but that activation at encoding may not precipitate the different levels of recall experience. PMID- 12725760 TI - Are action and perception in near and far space additive or interactive factors? AB - Functional imaging has revealed differential neural mechanisms underlying action directed toward near or far space. Because some neuropsychological studies of patients with visuospatial neglect failed to show near/far dissociations with perceptual tasks, we investigated whether action and perception elicit distinct cerebral representations in near and far space. We measured regional cerebral blood flow with positron emission tomography in normal volunteers who performed manual line bisection (action) and made line bisection judgments (perception). Stimuli were presented in near space or far space. Far space presentation enhanced activations in occipital cortex extending into the medial occipitotemporal cortex bilaterally, while near space presentation enhanced left occipital-parietal, parietal, and premotor cortex activity. Manual bisection activated the extrastriate, superior parietal, and premotor cortex bilaterally, while bisection judgments activated the right inferior parietal cortex, anterior cingulate, right dorsolateral prefrontal cortex, and extrastriate and superior temporal cortex bilaterally. The neural mechanisms responsible for the two tasks (perceptual/motor) were not differentially modulated by space of presentation. PMID- 12725761 TI - Evidence for axonal pathology and adaptive cortical reorganization in patients at presentation with clinically isolated syndromes suggestive of multiple sclerosis. AB - Previous work has suggested that functional reorganization of cortical areas might have a role in limiting the clinical impact of axonal pathology in patients with established multiple sclerosis (MS). Since there is evidence for irreversible tissue damage even in patients with early MS, we assessed, using functional MRI (fMRI) and a general search method, the brain pattern of movement associated cortical activations in patients at presentation with clinically isolated syndromes (CIS) suggestive of MS. To elucidate the role of cortical reorganization in these patients, we also investigated the extent to which the fMRI changes correlated with the extent of overall axonal injury of the brain. From 16 right-handed patients at presentation with CIS and 15 right-handed, age- and sex-matched healthy volunteers, we obtained: (1). fMRI (repetitive flexion extension of the last four fingers of the right hand), (2). conventional MRI scans, and (3). a new, unlocalized proton MR spectroscopy ((1)HMRS) sequence to measure the concentration of N-acetylaspartate of the whole brain (WBNAA). Compared to controls, patients with CIS had more significant activations of the contralateral primary somatomotor cortex (SMC), secondary somatosensory cortex, and inferior frontal gyrus. They also had significant decreased WBNAA concentration. Relative activation of the contralateral primary SMC was strongly correlated with WBNAA levels (r = -0.78, P < 0.001). This study shows that axonal pathology and functional cortical changes over a rather distributed sensorimotor network occur in patients at presentation with CIS suggestive of MS and that these two aspects of the disease are strictly correlated. This suggests that the increased functional recruitment of the cortex in these patients might have an adaptive role in limiting the clinical impact of irreversible tissue damage. PMID- 12725762 TI - The ERP omitted stimulus response to "no-stim" events and its implications for fast-rate event-related fMRI designs. AB - A major difficulty in fast-rate event-related fMRI experiments is the extensive overlap from adjacent trials in the stimulus sequence. One approach to address this problem is to include "no-stim" or "null" events as a trial type. These are randomized as if they were true stimulus events but no stimulus actually occurs. Assuming that no response is elicited by the null events, their time-locked average reflects only the averaged overlap. Thus, contrasting the averages for the other trial types versus the null-event average subtracts out the overlap, enabling the extraction of the response functions for these other trial types. ERP studies, however, have indicated that an endogenous brain response, the omitted stimulus response (OSR), can be evoked by a missing event in a stream of regularly occurring stimuli. To the extent that this response is elicited by null events in an event-related fMRI experiment, the null-event subtraction or contrast would falsely introduce the inverse of the OSR into the averaged responses to the other trial types. Using high-density ERP recordings, we investigated the effect of different percentages of omitted stimuli (11, 22, and 33%) on the auditory OSR at stimulus rates of one event per second or one event per 2 s. Significant OSRs were found for each percentage in the 1-s condition as well as in the 11% 2-s condition. The responses consisted of an early posterior negative wave (180-280 ms) followed by a larger anterior positive wave. These results have important implications for fast-rate fMRI designs, while also providing new data on the brain's response to omitted stimuli. PMID- 12725763 TI - Factors affecting the accuracy of near-infrared spectroscopy concentration calculations for focal changes in oxygenation parameters. AB - Near-infrared spectroscopy (NIRS) can be used to noninvasively measure changes in the concentrations of oxy- and deoxyhemoglobin in tissue. We have previously shown that while global changes can be reliably measured, focal changes can produce erroneous estimates of concentration changes (NeuroImage 13 (2001), 76). Here, we describe four separate sources for systematic error in the calculation of focal hemoglobin changes from NIRS data and use experimental methods and Monte Carlo simulations to examine the importance and mitigation methods of each. The sources of error are: (1). the absolute magnitudes and relative differences in pathlength factors as a function of wavelength, (2). the location and spatial extent of the absorption change with respect to the optical probe, (3). possible differences in the spatial distribution of hemoglobin species, and (4). the potential for simultaneous monitoring of multiple regions of activation. We found wavelength selection and optode placement to be important variables in minimizing such errors, and our findings indicate that appropriate experimental procedures could reduce each of these errors to a small fraction (<10%) of the observed concentration changes. PMID- 12725764 TI - Sexual dimorphism and asymmetries in the gray-white composition of the human cerebrum. AB - Using high resolution MRI scans and automated tissue segmentation, gray and white matter (GM, WM) volumes of the frontal, temporal, parietal, and occipital lobes, cingulate gyrus, and insula were calculated. Subjects included 23 male and 23 female healthy, right-handed subjects. For all structures, male volumes were greater than female, but the gray/white (G/W) ratio was consistently higher across structures in women than men. Sexual dimorphism was greater for WM than GM: most of the G/W ratio sex differences can be attributed to variation in WM volume. The corpus callosum, although larger in men, is less sexually dimorphic than the WM as a whole. Several regions demonstrate pair-wise asymmetries in G/W ratio and WM volume. Both the cingulate gyrus and insula exhibit strong asymmetries. The left cingulate gyrus is significantly larger than the right, and the G/W ratio of the left insula is significantly greater than that of the right. Although statistically significant sex differences and asymmetries are present at this level of analysis, we argue that researchers should be wary of ascribing cognitive functional significance to these patterns at this time. This is not to say, however, that these patterns are not important for understanding the natural history of the human brain, and its evolution and development. PMID- 12725765 TI - A comprehensive study of gray matter loss in patients with Alzheimer's disease using optimized voxel-based morphometry. AB - Voxel-based morphometry (VBM) has already been applied to MRI scans of patients with Alzheimer's disease (AD). The results of these studies demonstrated atrophy of the hippocampus, temporal pole, and insula, but did not describe any global brain changes or atrophy of deep cerebral structures. We propose an optimized VBM method, which accounts for these shortcomings. Additional processing steps are incorporated in the method, to ensure that the whole spectrum of brain atrophy is visualized. A local group template was created to avoid registration bias, morphological opening was performed to eliminate cerebrospinal fluid voxel misclassifications, and volume preserving modulation was used to correct for local volume changes. Group differences were assessed and thresholded at P < 0.05 (corrected). Our results confirm earlier findings, but additionally we demonstrate global cortical atrophy with sparing of the sensorimotor cortex, occipital poles, and cerebellum. Moreover, we show atrophy of the caudate head nuclei and medial thalami. Our findings are in full agreement with the established neuropathological descriptions, offering a comprehensive view of atrophy patterns in AD. PMID- 12725766 TI - Changes in cerebral morphology consequent to peripheral autonomic denervation. AB - Pure autonomic failure (PAF) is characterized by an acquired, selective, peripheral denervation of the autonomic nervous system. Patients with PAF fail to generate bodily states of arousal via the autonomic nervous system in response to physical or cognitive effort. We used voxel-based morphometry to test the hypothesis that changes in the morphology of brain regions involved in autonomic control would arise as a consequence to the longstanding absence of peripheral autonomic responses in PAF patients. Optimized voxel-based morphometry of structural magnetic resonance scans was used to test for regional differences in grey and white matter in 15 PAF patients and matched controls. There were no group differences observed in global measures of grey matter, white matter, or cerebrospinal fluid (CSF). We identified morphometric differences reflecting regional decreases in grey matter volume and concentration in anterior cingulate and insular cortices in PAF patients relative to controls. Morphometric differences in brainstem and subcortical regions did not reach statistical significance. Our findings suggest that peripheral autonomic denervation is associated with grey matter loss in cortical regions encompassing areas that we have previously shown are functionally involved in generation and representation of bodily states of autonomic arousal. The nature of these changes cannot be determined from morphometric analysis alone, but we suggest that they reflect experience-dependent change consequent upon loss of afferent input to brain regions involved in representation of autonomic states. PMID- 12725767 TI - How atypical is atypical language dominance? AB - Atypical, right-hemisphere language dominance is poorly understood. It is often observed in patients with brain reorganization due to lesions early in life. It can also be encountered in seemingly normal individuals. We compared the patterns of neural language activation in 7 individuals with left- and 7 with right hemisphere language dominance, none of whom had any evidence of brain lesions. We speculated that incongruencies in the activation patterns in atypical, right hemisphere language dominance could indicate a reorganized neural language system after undetected early brain damage. Functional magnetic resonance imaging analysis of brain activation during phonetic word generation demonstrated (1). no increased activation in the subdominant hemisphere in right compared to left language dominance, (2). a similar variability in the pattern of activation in both groups, and (3). a mirror reverse pattern of activation in right- compared to left-hemisphere dominant subjects. These findings support the view that in individuals with an unrevealing medical history right-hemispheric dominance constitutes a natural rather than an abortive variant of language lateralization. PMID- 12725768 TI - Activations related to "mirror" and "canonical" neurones in the human brain: an fMRI study. AB - In the macaque monkey ventral premotor cortex (F5), "canonical neurones" are active when the monkey observes an object and when the monkey grasps that object. In the same area, "mirror neurones" fire both when the monkey observes another monkey grasping an object and when the monkey grasps that object. We used event related fMRI to investigate where in the human brain activation can be found that reflects both canonical and mirror neuronal activity. There was activation in the intraparietal and ventral limbs of the precentral sulcus when subjects observed objects and when they executed movements in response to the objects (canonical neurones). There was activation in the dorsal premotor cortex, the intraparietal cortex, the parietal operculum (SII), and the superior temporal sulcus when subjects observed gestures (mirror neurones). Finally, activations in the ventral premotor cortex and inferior frontal gyrus (area 44) were found when subjects imitated gestures and executed movements in response to objects. We suggest that in the human brain, the ventral limb of the precentral sulcus may form part of the area designated F5 in the macaque monkey. It is possible that area 44 forms an anterior part of F5, though anatomical studies suggest that it may be a transitional area between the premotor and prefrontal cortices. PMID- 12725769 TI - Neural correlates of "hot" and "cold" emotional processing: a multilevel approach to the functional anatomy of emotion. AB - The neural correlates of two hypothesized emotional processing modes, i.e., schematic and propositional modes, were investigated with positron emission tomography. Nineteen subjects performed an emotional mental imagery task while mentally repeating sentences linked to the meaning of the imagery script. In the schematic conditions, participants repeated metaphoric sentences, whereas in the propositional conditions, the sentences were explicit questions about specific emotional appraisals of the imagery scenario. Five types of emotional scripts were proposed to the subjects (happiness, anger, affection, sadness, and a neutral scenario). The results supported the hypothesized distinction between schematic and propositional emotional processing modes. Specifically, schematic mode was associated with increased activity in the ventromedial prefrontal cortex whereas propositional mode was associated with activation of the anterolateral prefrontal cortex. In addition, interaction analyses showed that schematic versus propositional processing of happiness (compared with the neutral scenario) was associated with increased activity in the ventral striatum whereas "schematic anger" was tentatively associated with activation of the ventral pallidum. PMID- 12725770 TI - Optimal spline smoothing of fMRI time series by generalized cross-validation. AB - Linear parametric regression models of fMRI time series have correlated residuals. One approach to address this problem is to condition the autocorrelation structure by temporal smoothing. Smoothing splines with the degree of smoothing selected by generalized cross-validation (GCV-spline) provide a method to find an optimal smoother for an fMRI time series. The purpose of this study was to determine if GCV-spline of fMRI time series yields unbiased variance estimates of linear regression model parameters. GCV-spline was evaluated with a real fMRI data set and bias of the variance estimator was computed for simulated time series with autocorrelation structures derived from fMRI data. This study only considered fMRI experimental designs of boxcar type. The results from the real data suggest that GCV-spline determines appropriate amounts of smoothing. The simulations show that the variance estimates are, on average, unbiased. The unbiased variance estimates come at some cost to signal detection efficiency. This study demonstrates that GCV-spline is an appropriate method for smoothing fMRI time series. PMID- 12725771 TI - Cortical evidence of the perceptual backward masking effect on /l/ and /r/ sounds from a following vowel in Japanese speakers. AB - We examined the influence of stimulus duration of foreign consonant vowel stimuli on the MMNm (magnetic counter part of mismatch negativity). In Experiment 1, /ra/ and /la/ stimuli were synthesized and subjects were native Japanese speakers who are known to have difficulty discriminating the stimuli. "Short" duration stimuli were terminated in the middle of the consonant-to-vowel transition (110 ms). They were nevertheless clearly identifiable by English speakers. A clear MMNm was observed only for short-duration stimuli but not for untruncated long-duration (150-ms) stimuli. We suggest that the diminished MMNm for longer duration stimuli result from more effective masking by the longer vowel part. In Experiment 2 we examined this hypothesis by presenting only the third formant (F3) component of the original stimuli, since the acoustic difference between /la/ and /ra/ is most evident in the third formant, whereas F1 and F2 play a major role in vowel perception. If the MMNm effect depends on the acoustic property of F3, a stimulus duration effect comparable to that found with the original /la/ and /ra/ stimuli might be expected. However, if the effect is attributable to the masking effect from the vowel, no influence of stimulus duration would be expected, since neither stimulus contains F1 and F2 components. In fact, the results showed that the "F3 only" stimuli did not show a duration effect; MMNm was always elicited independent of stimulus duration. The MMN stimulus duration effect is thus suggested to come from the backward masking of foreign consonants by subsequent vowels. PMID- 12725772 TI - Linear regression with spatial constraint to generate parametric images of ligand receptor dynamic PET studies with a simplified reference tissue model. AB - For the quantitative analysis of ligand-receptor dynamic positron emission tomography (PET) studies, it is often desirable to apply reference tissue methods that eliminate the need for arterial blood sampling. A common technique is to apply a simplified reference tissue model (SRTM). Applications of this method are generally based on an analytical solution of the SRTM equation with parameters estimated by nonlinear regression. In this study, we derive, based on the same assumptions used to derive the SRTM, a new set of operational equations of integral form with parameters directly estimated by conventional weighted linear regression (WLR). In addition, a linear regression with spatial constraint (LRSC) algorithm is developed for parametric imaging to reduce the effects of high noise levels in pixel time activity curves that are typical of PET dynamic data. For comparison, conventional weighted nonlinear regression with the Marquardt algorithm (WNLRM) and nonlinear ridge regression with spatial constraint (NLRRSC) were also implemented using the nonlinear analytical solution of the SRTM equation. In contrast to the other three methods, LRSC reduces the percent root mean square error of the estimated parameters, especially at higher noise levels. For estimation of binding potential (BP), WLR and LRSC show similar variance even at high noise levels, but LRSC yields a smaller bias. Results from human studies demonstrate that LRSC produces high-quality parametric images. The variance of R(1) and k(2) images generated by WLR, WNLRM, and NLRRSC can be decreased 30%-60% by using LRSC. The quality of the BP images generated by WLR and LRSC is visually comparable, and the variance of BP images generated by WNLRM can be reduced 10% 40% by WLR or LRSC. The BP estimates obtained using WLR are 3%-5% lower than those estimated by LRSC. We conclude that the new linear equations yield a reliable, computationally efficient, and robust LRSC algorithm to generate parametric images of ligand-receptor dynamic PET studies. PMID- 12725773 TI - Spatial dependence of the nonlinear BOLD response at short stimulus duration. AB - Most functional magnetic resonance imaging studies use linear models to predict the measured response by convolution of an impulse response with the stimulus profile. Using very short visual presentation times (<2 s), deviation from the linear model in the measured BOLD data from the human brain was found for the response integral, amplitude, and width. In this study, high temporal and spatial resolution were used to quantify nonlinear effects and investigate the spatial dependence. Data at 4 Tesla showed at short stimulus duration a nonlinearity, i.e., deviation from a linear model, with an index up to 400%, whereas data at 7 Tesla exhibited a nonlinearity index up to 40%. The effect was more pronounced for response amplitude than for response area. A reduced width and sharpening of responses at shorter stimulus duration was also found. A voxel-based analysis of 7 Tesla data with 1.2 x 1.2 x 2 mm(3) resolution revealed a correlation between response onset and nonlinearity index. This suggests that the nonlinearity effects are a tissue-specific phenomenon and are likely to be more localized to the site of neuronal activity. The observed magnetic field dependence and the demonstrated nonlinearity in the response width support the hypothesis that the source of the nonlinearity at short stimulus duration has a considerable hemodynamic contribution. The nonlinearity was modeled as a "switch"-type initial hemodynamic response onset. Understanding these nonlinearities in the BOLD response is important for design and the analysis of rapid event-related fMRI experiments with brief stimulus presentations. PMID- 12725774 TI - Cortical activation to illusory shapes as measured with magnetoencephalography. AB - Spatiotemporal patterns of cortical activation during the perceptual grouping of elements to form illusory shapes were estimated using anatomically constrained magnetoencephalography. Subjects were shown an array of Kanizsa-style figures which were either aligned to form illusory squares or misaligned so that no illusory contour or shape was perceived. Differential activity is more pronounced in the right hemisphere. After a weakly significant modulation at approximately 110 ms in the occipital pole, a prominent peak appears at approximately 155 ms in the lateral occipital cortex. Modulation then appears to spread back from this location toward the occipital pole, as well as ventrally to involve ventral occipital and temporal cortices for the next 180 ms, eventually involving ventral orbitofrontal cortex at 325 ms. The prominent lateral occipital response is consistent with fMRI studies with similar stimuli which found activation in that region as well as in V3A, V4v, V7, and V8. Furthermore, the timing of this activation, after the occipital pole but before ventral temporal, is consistent with a putative role for this region in midlevel vision. The late ventral temporal response (235 ms) is centered in the lingual and fusiform areas implicated in object identification. The V1/V2 modulation at this time may reflect top-down modulation by lateral occipitotemporal and ventral temporal areas. PMID- 12725776 TI - In vivo recording of monophasic action potentials in awake dogs. AB - Assessment of cardiac repolarization in dogs in vivo is of interest in numerous experimental canine models. Previous studies have used monophasic action potentials (MAPs) to investigate repolarization in vitro and in vivo in anesthetized animal models. Therefore, an approach for recording MAPs in awake dogs without the interference of anesthesia is desirable. We describe an experimental technique to record MAPs in conscious dogs by means of conventional rubber introducers which are implanted into the internal jugular vein. Atrial as well as ventricular MAPs can be simultaneously measured without complications. Pacing thresholds are low and stable over time. Continuous MAP recordings of stable amplitude can be achieved from the same endocardial site for periods up to 1h. Antegrade and retrograde atrioventricular nodal conduction properties can be assessed. Programmed stimulation can be performed to simultaneously determine local refractory periods and MAP duration at cycle lengths from 500 to 200ms. In awake, unsedated dogs measuring MAPs via rubber introducers permits safe, long term recording of MAPs. Such recordings may be useful for safety pharmacologic studies in evaluating cardiovascular and noncardiovascular drugs with respect to their effects on repolarization. In various canine in vivo models including in vivo models of long QT syndrome, heart failures or sudden cardiac death, the present technique permits electrophysiologic measurements without the interference of anesthesia. PMID- 12725778 TI - Techniques for recording renal sympathetic nerve activity in awake, freely moving animals. AB - The methods described here allow recording of sympathetic nerve discharge in awake, freely moving animals, and follow a historical perspective of the different techniques developed over the years to record sympathetic discharge. The length of time each system is viable for recording varies from days to weeks. Also included are special hints for the surgical implantation of recording electrodes, types of recording electrodes and cables, as well as the minimum equipment necessary for recording sympathetic discharge. Lastly, a section on troubleshooting includes how to remove movement artifacts and extraneous noise, and minimize the destruction of leads common in recording in awake, freely moving animals. This article is written for the beginner or novice with an emphasis on what is needed when embarking on recording sympathetic nerve discharge in awake, freely moving animals. PMID- 12725777 TI - A radiotelemetry system for polysomnographic recordings in lambs. AB - The use of telemetry in biomedical research is rapidly increasing and is soon likely to be standard in research fields where prolonged recordings of multiple physiologic variables are necessary in freely moving animals. We describe our experience with a radiotelemetry system designed in our laboratory for polysomnographic recordings in newborn lambs. The system comprises eight channels: four channels for electromyographic activity of respiratory muscles, and one channel each for electrocorticogram, eye movement, electrocardiogram, and nasal flow. Using this system, we report our experience on the study of respiration and spontaneous apneas in the various states of consciousness, based on more than 500 h of recordings. Addition of radiotelemetry to our armamentarium has proven to be of invaluable help for the study of neonatal apneas and has now become routine in our laboratory. PMID- 12725779 TI - Electromyogram recordings from freely moving animals. AB - Electromyography can be used to record activity from sets of muscles in awake, freely moving animals using implanted intramuscular electrodes. As a tool, EMG has a wide range of applications ranging from inferring neural processes to analyzing movement. The amplitude of the rectified and filtered electromyogram (EMG) can be used as an indirect measure of muscle activity. Although it is often tempting to correlate the EMG with muscle force, the fact that force varies more with different activation strategies than with EMG estimates must be taken into account. The purpose of this article is to provide the researcher wishing to introduce the technique of recording EMGs from conscious animals using intramuscular electrodes with a step-by-step guide. It includes details on the manufacture of electromyograph electrodes, recording, and analysis considerations along with a section on solving common problems. For the sake of clarity, this article focuses on using the cat as a model and on the implantation of hindlimb muscles with intramuscular wire electrodes. However, the procedures can be adapted for use on other striated muscles and species. PMID- 12725780 TI - Single-unit recording in the lateral geniculate nucleus of the awake behaving monkey. AB - In recent years, recording neuronal activity in the awake, behaving primate brain has become established as one of the major tools available to study the neuronal specificity of the initiation and control of various behaviors. Primates have traditionally been used in these studies because of their ability to perform more complex behaviors closely akin to those of humans, a desirable prerequisite since our ultimate aim is to elucidate the neuronal correlates of human behaviors. A wealth of knowledge has accumulated on the sensory and motor systems such as vision, audition, and eye movements. For more demanding behaviors where the main focus has been on attention, recordings in awake primates have begun to yield valuable data on the centers of the brain that are reactive to different attributes of this behavior. As a result, various hypotheses of the origin and distribution of attentional effects have evolved. For instance, visual attentional effects have been described not only in the higher cortical area (V4) but also in areas earlier in the visual pathway which presumably involve a feedback mechanism in the latter region. Here we outline the ways in which we have successfully used these methods to make single-cell recordings in awake macaques to show how certain behavioral paradigms affect neurons of the thalamus (with emphasis on the lateral geniculate nucleus). As we have done with established techniques these methods can be readily adapted to incorporate most behaviors needed to be tested and allow recordings to be made in virtually any part of the brain. PMID- 12725781 TI - A lightweight microdrive for single-unit recording in freely moving rats and pigeons. AB - A design for an inexpensive and reliable subminiature microdrive for recording single neurons in the freely moving animal is presented. The Scribe microdrive is small and lightweight and has been used successfully to record in freely moving rats and pigeons. It would also be suitable for recording in mice. The device is simple and inexpensive yet allows for stable and precise manipulation of the recording electrodes. As a result it supports stable recordings conducted over long periods. Because the Scribe microdrive is a small-diameter device it is also suitable for multisite, multielectrode applications. Here we discuss the construction of the device and comment on its use in recording from freely moving rats and pigeons. PMID- 12725782 TI - In vivo electron paramagnetic resonance oximetry with particulate materials. AB - Electron paramagnetic resonance (EPR) methods can be used to study tissue pO(2) (PtO(2)) in anesthetized or awake animals (EPR oximetry). The method takes advantage of the fact that some paramagnetic materials have an EPR linewidth that is sensitive to the pO(2) in which the material is located. This article provides an overview of the method of EPR oximetry using implanted particulate materials as the sensors of pO(2). Characteristics of these materials are described to help the reader understand the factors involved in choosing the optimum particulate material. Examples of biological studies are included that show how EPR oximetry may be used on both awake and anesthetized animals. PMID- 12725783 TI - Brain temperature measurement and regulation in awake and freely moving rodents. AB - Temperature measurement and control are essential in most ischemia experiments. Hypothermia lessens ischemic brain injury whereas hyperthermia exacerbates it. A substantial number of ischemia studies rely solely on rectal temperature measurements during the insult. However, rectal temperature may not accurately reflect brain temperature especially during global ischemia. Furthermore, postischemic temperature changes are often inadequately monitored. Delayed cooling reduces injury, whereas delayed hyperthermia aggravates it. This review summarizes our experiences with core and brain telemetry probes to continually measure temperature in various ischemia models. Furthermore, we discuss methods to simultaneously measure and regulate temperature in the freely moving postischemic rodent, and the need for such control in ischemia research. PMID- 12725784 TI - Real-time cortical cerebral blood flow follow-up in conscious, freely moving rats by laser Doppler flowmetry. AB - This article describes a laser Doppler flowmetry (LDF) system that enables repeated measurements and thereby long-term followup of cortical cerebral blood flow (CBF) in awake and freely moving rats. The system consists of a specially designed flow probe adapter, a flow probe connector, and a LDF flow probe, which may thereby rotate through its own axis. During the experiment, the flow adapter is permanently mounted onto the rat's skull bone. A thin layer of skull bone is left intact at the site for cortical CBF measurements. The probe connector and the flow probe may be repeatedly detached and remounted to the adapter, which allows for cortical cerebral blood flow recording from exactly the same anatomical location. The laser Doppler flowmetry system enables stable cortical CBF recordings in the conscious rat while it moves freely in a bowl cage. PMID- 12725785 TI - An unsupervised automatic method for sorting neuronal spike waveforms in awake and freely moving animals. AB - The present study introduces an approach to automatic classification of extracellularly recorded action potentials of neurons. The classification of spike waveform is considered a pattern recognition problem of special segments of signal that correspond to the appearance of spikes. The spikes generated by one neuron should be recognized as members of the same class. The spike waveforms are described by the nonlinear oscillating model as an ordinary differential equation with perturbation, thus characterizing the signal distortions in both amplitude and phase. It is shown that the use of local variables reduces the problem of spike recognition to the separation of a mixture of normal distributions in the transformed feature space. We have developed an unsupervised iteration-learning algorithm that estimates the number of classes and their centers according to the distance between spike trajectories in phase space. This algorithm scans the learning set to evaluate spike trajectories with maximal probability density in their neighborhood. Following the learning, the procedure of minimal distance is used to perform spike recognition. Estimation of trajectories in phase space requires calculation of the first- and second-order derivatives, and integral operators with piecewise polynomial kernels were used. This provided the computational efficiency of the developed approach for real-time application as required by recordings in behaving animals and in human neurosurgical operations. The new method of spike sorting was tested on simulated and real data and performed better than other approaches currently used in neurophysiology. PMID- 12725787 TI - Expression cloning for the discovery of viral antigens and epitopes recognized by T cells. AB - Knowledge of the antigens that are recognized by virus-specific T cells can identify candidate subunit vaccine compounds. Viral antigens are frequently used in cross-sectional and longitudinal studies of the immune response in pathogenesis research. Peptide epitopes are required for the construction of fluorescent major histocompatibility complex-peptide tetramers, which are used to track specific T-cell responses. Expression cloning is a family of methods of antigen discovery that are particularly suitable for DNA viruses with large genomes. Libraries of viral nucleic acid are expressed in formats suitable for presentation to either CD4 or CD8 T cells. Pools of antigens representing fragments of viral open reading frames are loaded into cells expressing the necessary antigen processing and presentation machinery. Highly sensitive T-cell readouts such as lymphokine secretion, proliferation, and gene activation are used to detect active pools, which are then broken down in a reiterative process until a single active clone can be isolated and sequenced. These methods are most applicable if T-cell clones reactive with the whole virus can be obtained by in vitro restimulation or sampling of infected tissues. Alternative methods of antigen discovery are better suited for nonculturable viruses. Expression cloning methods are somewhat generic and are adaptable between infectious diseases, autoimmunity, and tumor immunology research projects. PMID- 12725788 TI - Identification of MHC class II-restricted tumor antigens recognized by CD4+ T cells. AB - CD4+ T cells play a central role in orchestrating host immune responses against cancer as well as autoimmune and infectious diseases. Identification of major histocompatibility complex (MHC) class II-restricted helper T peptides is important for development of effective vaccines. The lack of effective methods to identify such T-cell peptides is a major hurdle in the use of antigen-specific CD4+ T cells in cancer vaccines. Here we describe a genetic targeting expression system for cloning genes encoding for MHC class II-restricted tumor antigens recognized by tumor-reactive CD4+ T cells. Helper T peptides are subsequently identified by using synthetic peptides to test their ability to stimulate CD4+ T cells. PMID- 12725789 TI - Use of combinatorial peptide libraries for T-cell epitope mapping. AB - T lymphocytes play important roles not only in infectious diseases and autoimmunity, but also in immune responses against tumors. For many of these disorders, the relevant target antigens are not known. Designing effective methods that allow the search for T-cell epitopes is therefore an important goal in the areas of infectious diseases, oncology, vaccine development, and numerous other biomedical specialties. So far, the strategies used to examine T-cell recognition have been based largely on mapping T-cell epitopes with overlapping peptides from known proteins or with entire proteins, e.g., from a specific virus, bacterium, or human tissue. These approaches are tedious and have a number of limitations. It is, for example, almost impossible to isolate T cells that infiltrate an organ or infectious site and identify their specificity unless one already has a concept as to which antigens may be relevant. During recent years, a number of laboratories have developed less biased approaches that employ either the selection of putative T-cell epitopes based on the prediction of binding to certain major histocompatibilty complex (MHC) molecules and peptide or protein libraries that have been generated in expression systems, e.g. phage, or rely on combinatorial peptide chemistry. The latter technique has been refined by a number of laboratories including ours. Bead-bound or, preferably, positional scanning synthetic and soluble combinatorial peptide libraries allow the identification of T-cell epitopes within complex mixtures of proteins even for T cells that have been expanded from an organ infiltrate with a polyclonal stimulus. The practical steps that are involved in the latter method are described in this article. PMID- 12725790 TI - The use of HPLC-MS in T-cell epitope identification. AB - The hunt for T-cell epitopes is going on because hopes are set on such peptide sequences for diagnosis and vaccine development in the fight against infectious and tumor diseases. In addition to a variety of other techniques used in T-cell epitope identification, mass spectrometers coupled to microcapillary liquid chromatography have now become an important and sensitive tool in separation, detection, and sequence analysis of highly complex natural major histocompatibility complex (MHC) ligand mixtures. In this article, we review the basics of mass spectrometric techniques and their on-line coupling to microcapillary liquid chromatography (microcap-LC). Furthermore, we introduce current strategies for the identification of new T-cell epitopes using microcapillary liquid chromatography-mass spectrometry (microcap-LC-MS). PMID- 12725791 TI - T-cell epitope mapping using the ELISPOT approach. AB - The ELISPOT assay is particularly well suited to measure both clonal size and effector function of low-frequency antigen-specific T-cell populations directly ex vivo. Typically, an ELISPOT assay is performed with a freshly obtained sample (or cryopreserved sample) using less than 24h of culture. Additionally, this assay allows for the simultaneous analysis of hundreds of variables in parallel from a single tissue specimen. Because of these capabilities, this assay has found widespread use in the context of direct ex vivo immune diagnostic monitoring in humans. Herein we describe the rationale, methodology, and useful hints for performing T-cell epitope mapping using ELISPOT analysis, and review typical results of such an analysis. PMID- 12725792 TI - Mapping T cell epitopes by flow cytometry. AB - Epitope mapping by flow cytometry is a very modern approach that not only identifies T-cell epitopes but simultaneously allows for detailed analysis of the responding T-cell subsets including lineage, activation marker expression, and other markers of interest. The most frequently used approach is based on the identification of intracellular cytokines in secretion-inhibited activated T cells following stimulation with peptides or peptide pools. A more recently developed assay analyzes T-cell proliferation by measuring the decrease in carboxyfluorescein diacetate succinimidyl ester staining in proliferated cells. This article includes information on peptide configuration, a section on the design and efficient application of peptide pools, and working laboratory protocols for both assays. PMID- 12725793 TI - Use of HLA class II tetramers in tracking antigen-specific T cells and mapping T cell epitopes. AB - The highly specific interaction between major histocompatibility complex (MHC)/peptide and its cognate T-cell receptor (TCR) directs to the specificity of the T-cell response. Although the interaction affinity of individual MHC/peptide and TCR molecules is low, multimerization of the MHC/peptide complexes increases the avidity of their interaction with TCRs, and enables the complexes to be used as staining reagents for antigen-specific T cells. Different approaches are used to generate class I and class II tetramer reagents, and these reagents have been demonstrated to be useful in identifying CD8+ and CD4+ T cells, respectively. This article focuses on the uses of human lymphocyte antigen (HLA) class II tetramers in tracking antigen-specific T cells. Class II tetramers can also be used to map antigenic epitopes of known antigens through a novel peptide screening procedure known as tetramer-guided epitope mapping (TGEM). In this article, we provide a description of the methodological approach of using HLA class II tetramers in tracking and isolating T cells, and the use of class II tetramers in mapping T-cell epitopes. PMID- 12725794 TI - Bioinformatics tools for identifying class I-restricted epitopes. AB - The lack of simple methods to identify relevant T-cell epitopes, the high mutation rate of many pathogens, and restriction of T-cell response to epitopes due to human lymphocyte antigen (HLA) polymorphism have significantly hindered the development of cytotoxic T-lymphocyte (CTL) epitope-based or "epitope-driven" vaccines. Previously, CTL epitopes were mapped using large arrays of overlapping synthetic peptides. The large number of protein sequences available for mapping is now making this method prohibitively expensive and time-consuming. Bioinformatics tools such as EpiMatrix and Conservatrix, which search for unique or multi-HLA-restricted (promiscuous) T-cell epitopes and identify epitopes that are conserved across variant strains of the same pathogen, accelerate epitope mapping. These tools offer a significant advantage over other methods of epitope selection because high-throughput screening can be performed in silico, followed by confirmatory studies in vitro. CTL epitopes discovered using these tools might be used to develop novel vaccines and therapeutics for the prevention and treatment of infectious diseases such as human immunodeficiency virus, hepatitis C, tuberculosis, and some cancers. PMID- 12725795 TI - The use of bioinformatics for identifying class II-restricted T-cell epitopes. AB - An important step in the design of subunit vaccines is the identification of promiscuous T helper cell epitopes in sets of disease-specific gene products. Most of the epitope prediction models are based on HLA-II peptide binding, which constitutes a major bottleneck in the natural selection of epitopes. Here we describe a computer model, TEPITOPE, that enables the systematic prediction of promiscuous peptide ligands for a broad range of HLA binding specificity. We show how to apply the TEPITOPE prediction model to identify T-cell epitopes, and provide examples of its successful application in the context of oncology, allergy, and infectious and autoimmune diseases. PMID- 12725797 TI - In situ hybridization analysis of chemokines and chemokine receptors in the central nervous system. AB - The expression of a number of chemokines and chemokine receptors by cells resident in normal and pathological central nervous system (CNS) tissue has been characterized by in situ hybridization techniques. As a result, our understanding of the role of this cytokine family in neurobiology has been enhanced greatly. Specific methods for detecting chemokine and chemokine receptor mRNAs in situ vary with the number of these genes that have been characterized and encompass approaches widely utilized by other investigators characterizing cell-specific gene expression patterns. We describe methods that our laboratory has used successfully in characterizing chemokine and chemokine receptor expression in the CNS, focusing on protocols that utilize radiolabeled in vitro-transcribed riboprobes for detecting these transcripts. Because general dye-based histological staining methods do not readily differentiate astrocytes and microglia, specific immunohistochemical protocols are required for definitive localization of gene expression to these glial cell types. As such, methods that are compatible with the in situ hybridization procedure are included for staining astrocytes and microglia. PMID- 12725798 TI - Flow cytometric analysis of chemokine receptor expression on cerebrospinal fluid leukocytes. AB - Collection of cerebrospinal fluid (CSF) from the lumbar subarachnoid space is a routine procedure in clinical neurology, providing an opportunity to obtain hematogenous cells from the central nervous system environment in vivo. The ability to study individual cells in samples with low cell numbers has made flow cytometry an attractive method for studies of chemokine receptor expression on such cells. Several methodological variables such as staining temperature and cell isolation techniques may, however, influence the final outcome of the staining. In addition, low numbers of cells in the normal lumbar CSF, together with a tendency of CSF cells to decay rapidly after sampling, require meticulous handling of the samples. Here, we describe the methodology used in our laboratory to study chemokine receptor expression on cells in paired samples from peripheral blood and CSF using flow cytometry. PMID- 12725799 TI - Chemokine receptor binding and signal transduction in native cells of the central nervous system. AB - Chemokine receptors belong to the superfamily of seven-transmembrane-spanning, G protein-coupled receptors, and their expression by central nervous system cells is clearly documented. As this gene family has become the target of novel therapeutic development, the analysis of these receptors requires radioligand binding techniques as well as methods that entail assessing receptor stimulation of signal transduction pathways. Herein, we describe specific protocols for measuring radiolabeled chemokine binding to their cognate receptors on cultured glial cells as well as to receptors expressed in heterologous cell systems. Multiple downstream signaling pathways, including intracellular calcium influx and receptor-dependent kinase activation, are associated with chemokine receptor stimulation. Protocols for measuring these signaling events in chemokine-receptor expressing cells are also presented. PMID- 12725800 TI - Electrophysiological analysis of neuronal chemokine receptors. AB - Several studies have shown that neurons in the central and peripheral nervous systems express a variety of chemokine receptors (CKRs). Activation of these receptors can influence neuronal signaling by regulating synaptic transmission and neuronal excitability. This article presents electrophysiological methods that are currently used to study the normal and pathophysiological role for CKRs in the nervous system. Conventional electrophysiological methods such as patch clamp recording of isolated neurons, brain slices, and heterologous expression systems are described. In addition, single-cell reverse transcription-polymerase chain reaction is discussed as a technique that can be used in conjunction with patch-clamp recording to further investigate the molecular basis of neuronal CKR activation. PMID- 12725801 TI - Use of cocultured cell systems to elucidate chemokine-dependent neuronal/microglial interactions: control of microglial activation. AB - In order to understand processes involved in central nervous system inflammatory diseases, a critical appreciation of mechanisms involved in the control of immune function in the brain is needed. Microglial cells are watchful eyes for unusual events and detecting the presence of pathogens but are also alert to signals emanating from damaged neurons. Fractalkine (CX3CL1) is a chemokine which is expressed predominantly in the central nervous system, being localized on neurons, while its receptor, CX3CR1, is found on microglial cells. We have developed a strategy to investigate the role of this chemokine in neuronal microglia interactions. Because fractalkine is expressed both as a soluble and as a membrane-attached protein, we have established various protocols involving different levels of cell-to-cell communication. Three experimental systems were instituted, including (1) a conditioned medium transfer system in which no cell cell communication or contact is possible, (2) a transwell system that permits cell-contact-independent communication through diffusible soluble factors only, and (3) a coculture system allowing cell-to-cell communication via direct microglial-neuronal contacts. Using these in vitro cocultured systems, we have investigated the role of a soluble and/or cell-associated chemokine, such as fractalkine, in order to obtain insights into the role of glia-neuron interactions in cerebral inflammation. PMID- 12725802 TI - Chemokine-dependent mechanisms of leukocyte trafficking across a model of the blood-brain barrier. AB - Leukocyte transmigration across the blood-brain barrier (BBB) is a multistep process that can be mediated by chemokines. These low-molecular-weight chemoattractant proteins are secreted by cells within the central nervous system (CNS) in response to injury or on activation. Leukocytes transmigrate toward this chemokine gradient, crossing the BBB and gaining access to the CNS parenchyma. Depending on the chemokine, the nature of the insult, and the type of cell that transmigrates, the BBB integrity may be disrupted, leading to its increased permeability. Both the inflammation resulting from leukocyte transmigration and BBB perturbations contribute to CNS pathology. The mechanisms that mediate leukocyte transmigration and BBB disruption, as well as tissue culture models that are used to study leukocyte trafficking, are the focus of this review. PMID- 12725803 TI - Immunoneutralization of chemokines for the prevention and treatment of central nervous system autoimmune disease. AB - Chemokine-induced lymphocyte migration has long been hypothesized to regulate the appearance and continued presence of lymphocytes and monocytes in tissue-specific autoimmune diseases, including central nervous system autoimmune diseases such as multiple sclerosis. For instance, a large body of evidence points to the temporal association of chemokine expression with the appearance of T lymphocytes and monocytes/macrophages. Furthermore, experiments using mice with targeted mutations for chemokines have shown the importance of those molecules in the development of central nervous system autoimmune disease. We have hypothesized that temporal and spatial expression of chemokines is a key factor in the pathogenesis of experimental autoimmune encephalomyelitis and multiple sclerosis. To test our hypothesis we have employed the strategy of eliminating chemokine function by the passive transfer of chemokine-specific polyclonal antibodies. This approach has allowed us not only to test the function of chemokines in experimental autoimmune encephalomyelitis development, but also to ask questions about the roles of chemokines during disease progression. Moreover, this approach has allowed us to assess the efficacy of targeting chemokines and their receptors for treatment of ongoing disease. In the present report we summarize our experience using anti-chemokine administration for the prevention and treatment of experimental autoimmune encephalomyelitis as well as provide specific examples of how this approach is efficacious for disease treatment. PMID- 12725804 TI - Development and evaluation of pharmacological agents targeting chemokine receptors. AB - Chemokine receptors belong to one of the most pharmacologically exploited proteins, the G-protein-coupled receptors. Drugs that target these receptors make up greater than 45% of all known marketed medicines. Several excellent reviews published recently have concentrated on the biology, pathophysiology, and molecular mechanisms of action of the chemokines [C. Gerard, B.J. Rollins, Nat. Immunol. 2 (2001) 108; C.R. Mackay, Nat. Immunol. 2 (2001) 95; M. Thelen, Nat. Immunol. 2 (2001) 129] and the reader is directed toward them to gain a thorough understanding of the importance of this growing family of proteins. Although some background will be given here to aid in an understanding of the medical importance of chemokines, this review will focus on the rapid advances that have been made in identifying and characterizing chemokine receptor antagonists, by discussing their efficacy in animal models of disease as well as detailing their progression through human clinical trials. This approach is exemplified by specific reference to CCR1 and CCR5, which are the most advanced chemokine receptor programs. PMID- 12725805 TI - Food utilization efficiency in fifth instar larvae of Antheraea mylitta (Lepidoptera: Saturniidae) infected with Nosema sp. and its effect on reproductive potential and silk production. AB - Antheraea mylitta, a sericigenous insect of economical importance is often infected with an intracellular parasite of the genus Nosema. This pathogen is known to cause fatal pebrine disease and is considered as an important factor that strongly influences the development of the host. Larvae developed from the eggs laid by a female infected with Nosema sp. showed extended development period. The increment in the larval weight declined significantly in infected larvae in comparison to uninfected ones. Food consumption, digestion, relative consumption rate (RCR), efficiency of conversion of ingested food (ECI), efficiency of conversion of digested food (ECD), and relative growth rate (RGR) values declined significantly, but at the same time a significant increase in approximate digestibility (AD) was also observed. Silk production declined in infected larvae. Silk gland weight and shell weight also significantly declined following infection over uninfected larvae. The reproductive potential in adults declined significantly (P<0.001) with decrease in ovary weight (31.6%), fecundity (54.1%), and fertility (34.9%). Egg chorionation was also affected in adults, which developed from infected larvae. The maternal infection level in one generation (10.4 x 10(6) spores/female) decreased significantly in the next generation (8.0 x 10(6) spores/female). PMID- 12725806 TI - Nutritional requirements of Pochonia chlamydospora and ARF18, fungal parasites of nematode eggs. AB - Twenty carbohydrates (C), 18 nitrogen compounds (N), and 9 vitamins were examined for their effects on the growth and conidiation of the nematode-egg-parasitic fungi Arkansas Fungus 18 (ARF18, isolate 908) and Pochonia chlamydospora var. chlamydospora in solid and liquid cultures. Glycogen was the best, and inulin, D (+)-galactose, and soluble starch were good C sources for the growth of ARF18 in both solid and liquid cultures. ARF18 could not utilize alpha-cellulose in liquid; alpha-lactose and D-mannitol in solid; and D-(+)-xylose, L-(-)-sorbose, and D-(-)-ribose in both solid and liquid cultures. Casein was the best N source for ARF18 growth in both solid and liquid cultures and L-aspartic acid, DL glutamic acid, peptone, and L-histidine were good in solid culture. ARF18 could not utilize L-cystine and L-tyrosine in solid culture, and L-cystine, DL methionine, peptone, L-proline, and ammonium nitrate in liquid culture. Supplement of vitamins appeared to be unnecessary for ARF18 to grow. For P. chlamydospora var. chlamydospora growth, all test C sources, except L-(-) sorbose, alpha-cellulose, citric acid, and D-(+)-glucose, were good in both solid and liquid cultures. Most N compounds were good for P. chlamydospora var. chlamydospora growth with casein and peptone the best. Vitamins had limited effect on P. chlamydospora var. chlamydospora growth. P. chlamydospora var. chlamydospora conidiation was well supported by D-(-)-ribose, D-(-)-fructose, melibiose, and D-(+)-galactose as C sources and by L-aspartic acid, DL-glutamic acid, and L-arginine as N sources. Excluding myo-inositol from the medium containing all other test vitamins increased P. chlamydospora var. chlamydospora conidiation, while excluding pyridoxine appeared to reduce its conidiation. PMID- 12725807 TI - Entomophthora leyteensis Villacarlos & Keller sp. nov. (Entomophthorales: Zygomycetes) infecting Tetraleurodes acaciae (Quaintance) (Insecta, Hemiptera: Aleyrodidae), a recently introduced whitefly on Gliricidia sepium (Jaq.) Walp. (Fabaceae) in the Philippines. AB - Entomophthora leyteensis Villacarlos & Keller sp. nov., a species of Entomophthorales infecting the whitefly Tetraleurodes acaciae on Gliricidia sepium in the Philippines is described. Disease prevalence monitored weekly for 8 weeks indicated that the fungus could cause 8-31% infection within the whitefly population. Epizootics due to this fungus occurred in Inopacan, Leyte. Sampling live whitefly adults and dissecting them on glass slides for microscopic examination of fungal structures was found to give a better measure of prevalence than actual counts of infected insect cadavers. E. leyteensis is an important mortality factor for T. acaciae. Some speculations on the origin of the fungus are discussed here. PMID- 12725809 TI - Comparative pathogenesis of the Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus in noctuid hosts of different susceptibility. AB - Neonate larvae of the noctuid moth Spodoptera exigua were susceptible to an infection by Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus (HaSNPV). Biological activity (LD(50),ST(50)) of the virus was considerably reduced as compared to its activity in the homologous host, H. armigera. Pathogenesis was studied using a recombinant HaSNPV carrying a green fluorescent protein gene, which induces fluorescence in infected cells to mark infection. In larvae of H. armigera, fluorescence was pronounced in the fat body after 2.9 days post infection and could also be detected in several other tissues. In contrast, fluorescence was not observed in tissues of S. exigua until 9 days post infection and was restricted almost exclusively to cells of the ganglia. Examination of serial sections of wildtype HaSNPV-infected S. exigua-larvae revealed a similar pattern of tissue tropism. Apparently, HaSNPV does not undergo the usual steps in host invasion and infection in this insect species, but targets specifically to nervous tissue. PMID- 12725808 TI - Changes in circulating and tissue-infiltrating hemocyte parameters of European flat oysters, Ostrea edulis, naturally infected with Bonamia ostreae. AB - We assayed European flat oyster, Ostrea edulis, hemocyte parameters, circulating and tissue-infiltrating hemocyte densities, circulating hemocyte type distribution and lysosomal enzyme contents, to possibly relate these hematological parameters to Bonamia ostreae infection. Circulating hemocyte densities were not statistically different between infected and uninfected oysters. In contrast, the number of tissue-infiltrating hemocytes increased with infection intensity suggesting a recruitment process at the site of infection and a possibility for cells to migrate from circulatory system to connective tissues. Lysosomal enzymes were localized mainly in granulocytes both infected and uninfected, and mean of alpha-naphtyl butyrate esterase activity decreased with increasing B. ostreae infection level. The main response observed was a change in hemocyte type distribution between uninfected and infected oysters and greater tissue-infiltrating hemocytes with increased infections. These results suggest that the decrease of circulating granulocytes, and, consequently of some cell enzyme activities may be related with B. ostreae infection. PMID- 12725810 TI - Protozoan and intracellular fungal gut endosymbionts in Drosophila: prevalence and fitness effects of single and dual infections. AB - We report on the effect of single and mixed infections with two gut symbionts, trypanosomatids and the intracellular fungus Coccidiascus legeri, on the life history of their host, Drosophila melanogaster. We also provide the first report on the prevalence of C. legeri in natural populations of Drosophila. Prevalence overall was low (3.4%), and differed with host species, but persisted from the first to the second year of our survey. We documented delayed pupation in flies exposed to trypanosomatids, but larvae exposed to the fungus eclosed more quickly than controls. Larvae exposed to mixed infections pupated more slowly, but eclosed more quickly than controls. PMID- 12725811 TI - Diverse origins of tetracycline resistance in the honey bee bacterial pathogen Paenibacillus larvae. AB - Paenibacillus larvae is the causative agent of the important honey bee larval disease American Foulbrood (AFB). This pathogen has been treated in bee colonies by a single registered antibiotic, oxytetracycline (OTC), for fifty years. Recently, widespread resistance to OTC has been reported. In this study, the degree of antibiotic resistance was contrasted with DNA sequence variation for 125 P. larvae isolates collected in North America. Resistance was uncorrelated with bacterial haplotype, suggesting either that resistance has evolved multiple times in P. larvae or that resistance involves recent horizontal transfer via a non-genomic (e.g., plasmid or conjugal transposon) route. The recency of OTC resistance in P. larvae across this broad survey area underscores the need to manage foulbrood infections carefully and to monitor populations for resistance. PMID- 12725812 TI - The characterization of microsporidian isolates (Nosematidae: Nosema) from five important lepidopteran pests in Taiwan. AB - Microsporidian isolates from five lepidopteran pests-Spodoptera litura, Spodoptera exigua, Helicoverpa armigera, Plutella xylostella, and Pieris spp. were compared by spore morphology, infectivity to S. litura, Western-blot banding patterns, the sequences of small subunit rRNA gene (SSUrRNA sequence), and random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR). All the isolates could infect experimentally and multiply in the larvae of S. litura. The S. exigua isolate showed the highest virulence to the larvae of S. litura while the P. xylostella isolate showed the lowest. No significant differences either in spore morphology or in SSUrRNA sequences of these isolates were found. The SSUrRNA sequences of these isolates confirmed they are members of the genus Nosema. Based on the result of Western-blot hybridization with the rabbit anti Nosema spodopterae spore antiserum, three serotypes could be distinguished: N. spodopterae (S. litura isolate) and Pi. spp. isolate; S. exigua and H. armigera isolates; and P. xylostella isolate. The amplicons of RAPD-PCR with 60 primers yielded clear patterns that were selected and used for identification and also for phylogenic analysis of these five isolates. Based on analysis by the computer, isolates could be clearly divided into three groups that were coincident with the serotypes; therefore we suggest that N. spodopterae and isolates of Pi. spp., S. exigua, and H. armigera are more closely related in phylogenesis. In addition, in the amplification with the Nosema bombycis specific primer set, only DNAs from P. xylostella isolate and N. bombycis yielded amplicons. Therefore, we suggest that four isolates, excluding the P. xylostella isolate, are N. spodopterae, and the taxonomic position of P. xylostella isolate needs to be elucidated. PMID- 12725813 TI - Effects of mycotoxins, kojic acid and oxalic acid, on biological fitness of Lygus hesperus (Heteroptera: Miridae). PMID- 12725814 TI - Infectivity and reproductive potential of the Oswego strain of Heterorhabditis bacteriophora associated with life stages of the clover root curculio, Sitona hispidulus. AB - The infectivity and reproductive potential of the entomopathogenic nematode Heterorhabditis bacteriophora (Oswego strain), at different concentrations, was studied. Seventy to 80.0% mortality to late instar larvae of the clover root curculio, Sitona hispidulus, and 40.0-76.0% mortality to pupae, was observed at concentrations of 15-100 infective juveniles. There were no significant differences in mortality among nematode concentrations. LC(50) levels of 4.0 and 21.4 nematodes were determined for clover root curculio larvae and pupae, respectively. Nematodes did not cause significant mortality to adult or first instar clover root curculio. H. bacteriophora was able to complete its development and reproduce in 74.0-95.0% of clover root curculio late instar larvae and pupae. Reproductive potential in curculio larvae and pupae ranged from 0 to 7040 infective juveniles per host. Larvae exposed to 100 nematodes had a reproductive potential significantly higher than in those larvae exposed to 15 and 50 nematodes. Reproductive potential in pupae decreased with an increased nematode dose, indicating potential crowding effects. Host larval and pupal mass were positively correlated with nematode progeny production. PMID- 12725815 TI - Seasonal dynamics of endosymbiotic ciliates and nematodes in Dreissena polymorpha. AB - We report the results of a two-year study in the Svisloch River (Minsk, Belarus) on the dynamics of infection in Dreissena polymorpha by nematodes and three ciliate species Conchophthirus acuminatus, Ophryoglena sp., and Ancistrumina limnica. Although these endosymbionts were present in most of the samples, their prevalence and infection intensity differed significantly. C. acuminatus and A. limnica infection intensities in both years of the study had a maximum in summer and were positively correlated with water temperature. In contrast, Ophryoglena sp. and nematode infection intensities were considerably lower in summer versus winter and were negatively correlated with temperature. In the first long-term study to monitor the size and reproductive rate of C. acuminatus, we found that mean length was negatively correlated with temperature and that temperature was positively correlated with asexual reproduction, with a peak of cell division in April as water temperatures increased. PMID- 12725816 TI - Morphology and ultrastructure of a Pasteuria form parasitic in Tylenchorhynchus cylindricus (Nematoda). PMID- 12725817 TI - Evasion of encapsulation by the polyembryonic parasitoid Copidosoma floridanum is mediated by a polar body-derived extraembryonic membrane. PMID- 12725818 TI - Micellization and interfacial properties of alkyloxyethylene sulfate surfactants in the presence of multivalent counterions. AB - Alkyloxyethylene sulfates are a special class of surfactants that are unusually stable in the presence of multivalent counterions and are not as prone to precipitation as anionic surfactants without intermediate ethoxy groups in the molecule. However, formation of micelles, their structure, and the properties of monolayers of these surfactants exhibit very interesting and sometimes unexpected properties depending on the nature of the ions dissolved in the solution. This paper presents a brief overview of our recent efforts to reveal the nature of these properties, including some new results. We show that the strong binding of multivalent (and particularly trivalent counterions) triggers a sphere-to cylinder shape transition of the micelles and facilitates their further growth, even at very low ionic strength. The properties of surfactant monolayers are coupled to those of the micelles in the bulk and are governed also by multivalent counterion binding. The effect of multivalent counterions on the aggregation and structure formation in anionic surfactant solutions has both fundamental and practical importance. PMID- 12725819 TI - Oscillating laminar electrokinetic flow in infinitely extended circular microchannels. AB - This article addresses the problem of oscillating laminar electrokinetic liquid flow in an infinitely extended circular microchannel. Based on the Debye-Huckel approximation for low surface potential at the channel wall, a complex variable approach is used to obtain an analytical solution for the flow. The complex counterparts of the flow rate and the current are linearly dependent on the pressure gradient and the external electric field. This property is used to show that Onsager's principle of reciprocity continues to be valid (involving the complex quantities) for the stated problem. During oscillating pressure-driven flow, the electroviscous effect for a given value of the normalized reciprocal electrical double-layer (EDL) thickness is observed to attain a maximum at a certain normalized frequency. In general, an increasing normalized frequency results in a reduction of EDL effects, leading to (i). a volumetric flow rate in the case of streaming potential approaching that predicted by the theory without EDL effects, and (ii). a reduction in the volumetric flow rate in the case of electroosmosis. PMID- 12725820 TI - Oscillating laminar electrokinetic flow in infinitely extended rectangular microchannels. AB - This paper has addressed analytically the problem of laminar flow in microchannels with rectangular cross-section subjected to a time-dependent sinusoidal pressure gradient and a sinusoidal electric field. The analytical solution has been determined based on the Debye-Huckel approximation of a low surface potential at the channel wall. We have demonstrated that Onsager's principle of reciprocity is valid for this problem. Parametric studies of streaming potential have shown the dependence of the electroviscous effect not only on the Debye length, but also on the oscillation frequency and the microchannel width. Parametric studies of electroosmosis demonstrate that the flow rate decreases due to an increase in frequency. The obtained solutions for both the streaming potential and electroosmotic flows become those for flow between two parallel plates in the limit of a large aspect ratio. PMID- 12725821 TI - Adsorptive removal of methylene blue from colored effluents on fuller's earth. AB - The adsorption behavior of methylene blue (MB) on four fuller's earth (FE) samples of varying compositions was investigated using a spectrophotometric technique to obtain information on the color removal. The distribution coefficient (K(D)) increased with an increase in the initial concentration (C(0)) of the dye, attained a maximum value, and decreased again at higher initial concentrations. Dye solutions became colorless for a C(0) value corresponding to maximum K(D). A progressively increased flocculation behavior in the clay suspensions was observed and the maximum value of K(D) corresponds to optimum flocculation of the clay. The K(D) values were found to decrease exponentially after of the solution again became colored while the amount adsorbed increased with an increase in the initial concentration of MB. Only adsorption data obtained for this region could be defined by adsorption isotherm equations. The shifts of the C(0) values corresponding to K(D)(max) toward higher concentrations were correlated with the composition of FE samples by using XRF, XRD patterns, and SEM images. The influence of temperature on MB adsorption was also studied and thermodynamic parameters were calculated. PMID- 12725822 TI - Analytical investigation of the self-desorption of the oligomers of mixtures of a polydisperse ethoxylated surfactant with sodium dodecylsulfate from a silica/water interface. AB - The adsorption isotherms onto a hydrophilic silica of mixtures of sodium dodecylsulfate (SDS) and of all the oligomers of a polydisperse nonylethylene glycol n-dodecyl ether (C(12)E(9)) surfactant were determined using a high performance liquid chromatography (HPLC) technique. Incorporation of the anionic surfactant to the negatively charged silica surface is favored by the adsorption of the nonionic surfactant. Comparison between the adsorption isotherms of mixtures of SDS with a monodisperse C(12)E(9) and a polydisperse C(12)E(9) shows that the adsorption of SDS at the silica/water interface is stronger with the latter material than with the former in a large surface coverage domain. The composition of the surface aggregates and the variation of the oligomer distribution in these aggregates were determined. The previously described phenomena called self-desorption which was observed for the global C(12)E(9) and SDS surfactant mixtures was confirmed: increasing the total concentration at a fixed surfactant ratio induces at high concentration a desorption of the anionic surfactant and all of the less polar oligomers from the solid/water interface. An interpretation scheme is proposed which assumes that the interaction of SDS is larger with the less polar oligomers than with the polar ones. The self desorption effect could then be considered as the consequence of the polydispersity of the nonionic surfactant and to the net repulsion interaction between SDS and the silica surface as the mole fraction of SDS in the surfactant mixture increases. PMID- 12725824 TI - The use of monodispersed colloids in the polishing of copper and tantalum. AB - The properties of abrasive particles play a significant role in chemical mechanical polishing (CMP) of metal and dielectric films in semiconductor device manufacturing. This study investigates the effects of the particle size, shape, and hardness of abrasives on the polishing of copper and tantalum films in the presence of different slurry chemistries. Well-defined dispersions of uniform particles, including spherical silica of varying diameters, hematite of different shapes, and hematite cores coated with silica of different thicknesses, were prepared and used to polish blanket films of Cu and Ta. It was shown that the total surface area of the solids in the slurry controlled the rate of material removal by pure silica for both Cu and Ta, while the surface quality of the polished films was better when higher silica content was used. The shape or the aspect ratio of hematite particles had a minor effect on the removal rate. In contrast, when hematite particles coated with silica were employed in the polishing of Cu and Ta, the polish rate decreased with increasing thickness of the shell. PMID- 12725823 TI - Ion exchange of Pb(2+), Cu(2+), Fe(3+), and Cr(3+) on natural clinoptilolite: selectivity determination and influence of acidity on metal uptake. AB - In the present study ion exchange of Pb(2+), Cu(2+), Fe(3+), and Cr(3+) on natural Greek clinoptilolite was examined in terms of selectivity toward the above heavy metals in single- and multicomponent solutions in batch systems. Also examined are the influence of clinoptilolite on solution acidity and the effect of acidity on the ion exchange process. Clinoptilolite increases solution acidity due to the exchange of H(+) cations with the cations initially present in its structure. H(+) cations should be considered as competitive ones in ion exchange processes, and consequently ion exchange of metals is favored at high acidity values. Cu(2+) and Cr(3+) are the most sensitive cations with respect to acidity. Selectivity determination demonstrates that the selectivity at total concentration 0.01 N and acidity 2 in both single- and multicomponent solutions is following the order Pb(2+)>Fe(3+)>Cr(3+) > or =Cu(2+). This order is set since the first days of equilibration. However, Cu(2+) shows remarkable changes in selectivity and generally its uptake and selectivity are increasing with time. On the other hand selectivity in single metal solutions where acidity is not adjusted is following the order Pb(2+)>Cr(3+)>Fe(3+) congruent with Cu(2+). PMID- 12725825 TI - The influence of PEO/poly(vinyl phenol-co-styrene sulfonate) aqueous complex structure on flocculation. AB - Colloidal suspensions were flocculated with complexes formed from high molecular weight polyethylene oxide (PEO) and a cofactor. Poly(vinyl phenol-co-potassium styrene sulfate) (PKS) or poly(styrene-co-styrene sulfonate) (PS-co-SSS) copolymers were used as the cofactors for this work. The larger the PEO/cofactor complex species, the better the initial flocculation. Factors such as increasing temperature or ionic strength that gave smaller complexes also gave poorer flocculation. Cofactor performance was sensitive to the balance of hydrophobic phenolic groups and hydrophilic styrene sulfonates. If there are too few phenolic groups, the PEO/PSK complexes are large but are too weak to give shear-resistant flocs, whereas complexes formed with high phenolic content PSK are relatively small, giving poorer flocculation but more shear-resistant flocs. Both phenyl and phenol groups are effective as the hydrophobic component in the cofactor. The hydrogen-bonding potential of phenolic cofactors does not seem to offer much advantage relative to phenyl groups. A crucial step in the flocculation is the adsorption of PEO/cofactor complex onto the target colloids. Thus, flocculation is sensitive to the target colloid surface chemistry. Positively charged precipitated calcium carbonate and surfactant-free polystyrene latex are particularly easy to flocculate because adsorption is driven by electrostatic and hydrophobic interactions, respectively. By contrast, the latex coated with hydrophilic poly(N-isopropylacrylamide) (PNIPAM) does not flocculate because the PEO/cofactor complex does not bind to PNIPAM. Finally, the flocculation of highly negatively charged, dextran sulfate coated calcium carbonate seems to be stimulated by the presence of soluble calcium ions that make the complex less soluble and more likely to adsorb. PMID- 12725826 TI - Latex particle size distribution by dynamic light scattering: novel data processing for multiangle measurements. AB - Multiangle dynamic light scattering (DLS) provides a better estimate of particle size distributions (PSD) than single-angle DLS. However, multiangle data treatment requires appropriate weighting of each autocorrelation measurement prior to calculation of the PSD. The weighting coefficients may be directly obtained from (i). the autocorrelation baselines or (ii). independent measurement of the average light intensity by elastic light scattering. However, the propagation of errors associated with such procedures may intolerably corrupt the PSD estimate. In this work, an alternative recursive least-squares calculation is proposed that estimates the weighting coefficients on the basis of the complete autocorrelation measurement. The method was validated through a numerical example that simulates the analysis of a polystyrene latex with a bimodal PSD and with "measurements" taken at 10 detection angles. The ill-conditioned nature of the problem determines that the "true" PSD cannot be recovered, even in the absence of errors. A sensitivity analysis was carried out to determine the effect of errors in the weighting coefficients on the PSD recoveries. PMID- 12725827 TI - Preparation and electrochemistry of azobenzene self-assembled monolayers on gold- long range tunneling and end-group hydrogen bonding effect. AB - A series of thiol-functionalied azobenzene derivatives (RAzoCnSH: R=H for n=3-6, abbreviated as AzoCnSH; R=CH(3)CONH for n=4, abbreviated as aaAzoC4SH) on gold electrodes were prepared and their self-assembly and electrochemical properties were studied by cyclic voltammetry. They all formed uniform and reproducible self assembled monolayers (SAMs) on gold and showed well-behaved voltammetric responses in aqueous solution. Both the length of the alkyl chain spacer and the H-bonding of the end acetamino group had effects on the stability and the electrochemical kinetics of the SAMs, and the effect of the H-bonding was dominant. The surface coverage of the SAMs (AzoCnSH) is gradually increased with an increase of the alkyl chain spacer length, whereas the presence of the terminal acetamino group leads to a greater increase of the surface coverage. At a low scan rate, voltammetric responses corresponding to an irreversible two electron, two-proton reduction/oxidation of the trans-azobenzene redox center were obtained in the range of +300 mV and -800 mV, which exhibited very large peak-to-peak splitting. At a high scan rate of 500 mV/s, two steps of reversible one-electron, one-proton reduction/oxidation corresponding to the cis-isomer in azobenzene-thiol SAMs (n is odd) was clearly observed between +300 and -200 mV. The apparent electron-transfer rate is decreased with increasing distance between the azobenzene redox center and the gold electrode. The existence of the end acetamino group which restricted the conformational change during the redox process also led to a decrease of the standard rate constant, and this restriction effect is more predominant than the distance effect. PMID- 12725828 TI - Dynamic oscillatory behavior of a linear viscoelastic fluid bounded by an ideal linear viscoelastic membrane in torsional flow. AB - In rotational oscillatory rheological measurement techniques involving the plate plate and cone-plate geometries, the interface between the measured liquid and the ambient atmosphere is sheared to the same extent as the liquid sample. In this paper, we look at the influence of a rheologically distinct lateral surface on the measured properties of the liquid and surface system when the surface is dynamically coupled to the bulk fluid. Inertia is taken into account, thus allowing for nonquasi-static velocity profiles in the massless surface film itself. PMID- 12725829 TI - A generalized analytical model for the elastic deformation of an adhesive contact between a sphere and a flat surface. AB - A new method to calculate the elastic deformation of a sphere on a flat surface is presented. The model considers the influence of short-range as well as long range attractive forces both inside and outside the actual contact area. In contrast to earlier models, this theory describes the nature of these deformations in the intermediate regime between the so-called JKR and DMT limits by simple analytic expressions. Equations for the calculation of the contact radius, the deformation, and the pressure distribution are given. In all equations, the critical force that might vary between the limiting values found in the DMT and the JKR model acts as transition parameter. PMID- 12725830 TI - Praseodymium(III)-substituted bismuth titanate thin-film generation using metallo organic precursors with an M-O-C skeleton and sol-gel technique and employing 4f 4f transition spectra as a probe to explore kinetic performance. AB - A hetero-trimetallic lanthanide-substituted bismuth titanate (BLT, where lanthanide is praseodymium) with stoichiometry Pr(0.75)Bi(3.25) Ti(3)O(12) has been obtained as both highly homogenized crystalline and amorphous thin films using three different BLT precursors: (i). precursor A (Pr(OC(3)H(7)(i))(3),Bi(OOCCH(3))(3),Ti(OC(3)H(7)(i))(4)); (ii). precursor B (Pr(OC(3)H(7)(i))(2)(acac),Bi(OOCCH(3))(3),Ti(OC(3)H(7)(i))(3)(acac)); and (iii). precursor C (Pr(OC(3)H(7)(i))(2)(acac),Bi(OOCCH(3))(3),Ti(OC(3)H(7)(i))(2)(acac))(2). These three BLT precursors (A, B, C) are prepared by reacting constituent monometallic precursors in the desired stoichiometry and by employing controlled acidic hydrolysis via the sol-gel method. Paramagnetic Pr(III), being a f(2) ion, gives characteristic 4f-4f transition bands (3H(4)-->3P(2), 3H(4)-->3P(1), 3H(4)- >3P(0), and 3H(4)-->1D(2)) in the visible region, the intensities of which have been found to be highly sensitive to even minor changes in the immediate coordination around Pr(III), occurring as a result of the progress of polycondensation reactions of three multicomponent BLT sols. We have used the changes with time in the intensities (represented by oscillator strengths of these four 4f-4f bands) and the magnitude and variation of the spectral parameters evaluated from the observed spectra with a view toward monitoring the sol-gel reactions of BLT precursors A, B, and C. 4f-4f transition spectra of the aliquots, withdrawn from the hydrolyzing A, B, and C sols at different time intervals, represent the changes occurring in the Pr(III) environment with the progress of sol-gel hydrolysis of BLT, and are used to investigate the kinetic performance in hydrolysis of the three precursors. Kinetics of hydrolysis of precursors A, B, and C indicate that all four f-f transition bands of Pr are almost equally sensitive to precursor hydrolysis in the order A>B>C. PMID- 12725831 TI - Zeta potential measurement of calcium carbonate. AB - The problem of scaling, which one finds in industrial heat exchangers, particularly in atmospheric coolers in nuclear power stations, depends on calcium carbonate deposits from fresh water. To better understand this phenomenon, we have examined the eventual implication of superficial electric charge of precipitated crystal nuclei. After a bibliographical review showing a fundamental divergence from already published results, this paper describes an experimental plant to measure the zeta potential in controlled conditions of thermodynamic equilibrium, oversaturation, or undersaturation of a CaCO(3)-H(2)O-CO(2) system taking into account simultaneously the three phases: gas, liquid, and solid. The zeta potential is measured by a crystalline-plug method with calcite or aragonite crystals. The potential cancels at thermodynamic equilibrium and is always negative for other conditions, in particular for oversaturation where the possibility of scaling exists. The analysis of these results suggests that the potential determining ions of the system are Ca(2+) and HCO(-)(3). PMID- 12725832 TI - Effect of two-step sol-gel reaction on the mesoporous silica structure. AB - In the present study, we investigated the effects of two-step sol-gel reaction by abrupt pH change on the SBA-15 and mesocellular silica foams (MCF). Mesoporous silica was fabricated by using triblock copolymer templates (poly(ethylene oxide) and poly(propylene oxide)). The prepared silica structure was characterized by X ray diffraction, transmission electron microscopy, and N(2) sorption experiment. Specifically, we prepared SBA-15 with long-range two-dimensional hexagonal arrangement of 3 to 6-nm feature spacing and MCF with larger pores of a few tens of nanometers. The pore size and ordering were influenced by pH change in a two step sol-gel reaction and the concentration of organic solvent. Although well ordered hexagonal arrangement of mesopores was prevalent in acidic conditions, the materials synthesized by a single-step reaction in neutral or basic conditions possessed gel-like structure without mesopores. However, the present two-step reaction (low pH sol-gel reaction followed by high pH reaction) not only produced mesoporous materials but also provided controllability of the pore size. In particular, mesoporous structures with pore sizes as large as those of MCF were successfully fabricated by the two-step reaction without using organic swelling agents. As expected, when xylene was added as a swelling agent, the pore size increased with the xylene/copolymer weight ratio. PMID- 12725833 TI - Plastic deformation in cake consolidation. AB - Cake filtration has a variety of applications in wastewater treatment by solid/liquid separation processes. In order to obtain the physical properties of the filter cake, a Compression-Permeability Cell (C-P Cell) can be used to examine the constitutive relationships among the solid compressive stresses, porosity and the specific filtration resistance. In this paper, a mathematical model is proposed to study the transient distribution of stresses, strains and void ratios in the confined cake of a C-P Cell under constant axial loads for dewatering. Simulations have been carried out using a plasticity model and the sensitivity analysis has revealed the possible contribution a finite plasticity can bring about the two types of cake examples with either medium or high compressibility. PMID- 12725834 TI - On the relationship between the preparation method and the physicochemical and catalytic properties of the CoMo/gamma-Al(2)O(3) hydrodesulfurization catalysts. AB - A series of CoMo/gamma-Al(2)O(3) catalysts have been prepared using various methodologies. One of them (EDF) was prepared by depositing the Mo species on the support via the equilibrium deposition filtration (EDF) technique and then the Co species by dry impregnation. Another catalyst (co-EDF) was prepared by depositing the Co and Mo species simultaneously via EDF. A third catalyst (co-WET) was prepared by depositing Mo and Co species simultaneously using the wet impregnation method. The fourth catalyst (WET) was prepared by depositing the Mo species through wet impregnation and then the Co species by dry impregnation. Finally, the fifth catalyst (s-DRY) was prepared by mounting the Mo species through successive dry impregnations and then the Co species by dry impregnation. In all cases the Mo and Co content was identical, giving a Co/(Co+Mo) ratio equal to 0.13. These catalysts were characterized using various physicochemical techniques (BET, NO chemisorption, DRS, LRS, TPR, and XPS), and their catalytic activity for the hydrodesulfurization of thiophene was determined. The trend observed for the HDS activity (namely, EDF>co-EDF>co-WET>s-DRY>WET) is attributed to similar trends observed for both the fraction of well-dispersed octahedral cobalt in the oxidic precursors and the concentration of the edge sulfur vacancies formed on the active phase of the sulfided samples. The EDF and co-EDF catalysts exhibited relatively low hydrogenating activity. The maximum HDS activity, achieved over the EDF catalyst, suggested the most suitable preparative strategy for the preparation of very active and less hydrogen-demanding CoMo/gamma-Al(2)O(3) HDS catalysts. PMID- 12725835 TI - Surface reactions kinetics between nanocrystalline magnetite and uranyl. AB - Magnetite is the most important end member of iron corrosion products under reducing environment, which is the condition expected in a deep geological high level radioactive waste disposal. Nanocrystalline magnetite was synthesized in the laboratory and its physicochemical properties were analyzed in detail. The kinetics of the adsorption of U(VI) and the kinetics of the actinide reduction to a lower oxidation state, in presence of the oxide, were studied by means of batch sorption techniques and X-ray photoelectron spectroscopy (XPS) analysis. The results showed that the uranium sorption and reduction processes on the magnetite surface have very fast kinetics (hours), the reduction process being triggered by sorption. XPS measurements showed that the speciation of uranium at the surface does not show significant changes with time (from 1 day to 3 months), as well as the quantity of uranium detected at the surface. The surface speciation depended on the initial pH of the contact solution. Considering that the Eh of equilibrium between magnetite and the solution, under our experimental conditions, is slightly positive (50-100 mV), the uranium reduction would also be thermodynamically possible within the liquid phase. However, the kinetics of reduction in the liquid occur at a much slower rate which, in turn, has to depend on the attainment of the magnetite/solution equilibrium. The decrease of uranium in solution, observed after the uranyl adsorption stage, and particularly at acidic pH, is most probably due to the precipitation of U(IV) formed in the solution. PMID- 12725836 TI - Rupture energy and wetting behavior of pendular liquid bridges in relation to the spherical agglomeration process. AB - A novel micro force balance (MFB) is used to investigate the rupture energy of a silicon oil liquid bridge formed in water between two glass particles of either the same or dissimilar surface energy. Rupture energies are integrated from force curves and compared with the models proposed by Simons et al. (Chem. Eng. Sci. 49 (1994) 2331) and Pitois et al. (Eur. Phys. J. B 23 (2001) 79). The latter showed slightly better agreement to the experimental data. Glass ballotini ( approximately 100 microm diameter) are either silanized, in order to increase their wettability toward the oil binder, or kept untreated. Results showed how the interaction between the binder and the particle influences the geometry, the capillary pressure, the force, and the rupture energy of the liquid bridge. Higher values of force and liquid bridge energy were measured between particles characterized by higher interaction (silanized-silanized configuration). A thermodynamic approach to the evaluation of the energy stored in a liquid bridge is also proposed. The mechanical work done to stretch apart the liquid bridge is evaluated as the difference of internal and hysteresis energy between the initial and the rupture configuration of the bridge. This approach showed good agreement with the experimental data only for liquid bridges formed between silanized and untreated glass particles. PMID- 12725837 TI - Dynamics of surfactant sorption at the air/water interface: continuous-flow tensiometry. AB - Dynamic interfacial tensiometry, gauged by axisymmetric drop shape analysis of static drops or bubbles, provides useful information on surfactant adsorption kinetics. However, the traditional pendant-drop methodology is not readily amenable to the study of desorption kinetics. Thus, the question of sorption reversibility is difficult to assess by this technique. We extend classical pendant/sessile drop dynamic tensiometry by immersing a sessile bubble in a continuously mixed optical cell. Ideal-mixed conditions are established by stirring and by constant flow through the cell. Aqueous surface-active-agent solutions are either supplied to the cell (loading) or removed from the cell by flushing with water (washout), thereby allowing study of both adsorption and desorption kinetics. Well-mixed conditions and elimination of any mass transfer resistance permit direct identification of sorption kinetic barriers to and from the external aqueous phase with time constants longer than the optical-cell residence time. The monodisperse nonionic surfactant ethoxy dodecyl alcohol (C(12)E(5)), along with cationic cetyltrimethyl ammonium bromide (CTAB) in the presence of added salt, adsorbs and desorbs instantaneously at the air/water interface. In these cases, the experimentally observed dynamic-tension curves follow the local-equilibrium model precisely for both loading and washout. Accordingly, these surfactants below their critical micelle concentrations (CMC) exhibit no detectable sorption-activation barriers on time scales of order a min. However, the sorption dynamics of dilute CTAB in the absence of electrolyte is markedly different from that in the presence of KBr. Here CTAB desorption occurs at local equilibrium, but the adsorption rate is kinetically limited, most likely due to an electrostatic barrier arising as the charged surfactant accumulates at the interface. The commercial, polydisperse nonionic surfactant ethoxy nonylphenol (NP9) loads in good agreement with local-equilibrium theory but shows deviation from the theoretical washout curve, presumably due to slow desorption of solubilized but otherwise water insoluble components. The polymeric nonionic triblock copolymer Pluronic exhibits almost complete irreversible adsorption at the air/water interface over a molecular-weight range from 3 to 14 kDa. Similar irreversible dynamic behavior is observed for adsorption/desorption of the protein bovine serum albumin (BSA) from dilute aqueous solutions at the air/water interface. The new continuous-flow tensiometer (CFT) is a simple, yet powerful, tool to investigate sorption dynamics at fluid/fluid interfaces, especially for larger molecular weight surface-active agents that exhibit significant hindrance to desorption. PMID- 12725838 TI - Comparison of various models describing the adsorption of surfactant molecules capable of interfacial reorientation. AB - The thermodynamic model for describing the adsorption of surfactant molecules in different adsorption states, the reorientation model, is reconsidered on a more rigorous level. The resulting model equations are used to describe experimental surface pressure data published in the literature. The new model proposed contains three physical parameters and describes the experimental dependencies Pi(c) for oxethylated alcohols very accurately. PMID- 12725839 TI - Neutron reflection and small-angle neutron scattering studies of a fluorocarbon telomer surfactant. AB - Dilute aqueous phase behavior of a novel tris(hydroxymethyl)acrylamidomethane (THAM)-derived telomer bearing a perfluorohexyl hydrophobic chain, F6THAM6, has been investigated. Fluorinated polyhydroxy surfactants of this kind find use in emerging biomedical applications. Neutron reflection (NR) and drop volume surface tension (DVT) methods have been used to determine the critical micelle concentration (cmc=4.7 x 10(-4) mol x dm(-3)) and surface adsorption parameters (at the cmc NR gives a molecular area a(cmc)=67.4 and 62 A(2) and surface excess gamma(cmc)=2.46 x 10(-6) mol x m(-2)). The aggregation structures were determined by small-angle neutron scattering (SANS), indicating globular (polydisperse spheres) micelles of radius approximately 30 A are present. These findings are compared with literature on surfactants with related structures, to identify how the unusual molecular structure of F6THAM6 affects surfactant properties. PMID- 12725841 TI - Polymer surfactant kinetics using surface plasmon resonance spectroscopy dodecyltrimethylammonium chloride/polyacrylic acid system. AB - Kinetics of polymer surfactant interactions and the effect of surfactant binding on the conformational dynamics of the polymer were explored in this work using surface plasmon resonance spectroscopy. Polyacrylic acid was modified with thiol to varying degrees so as to force the polymer to form different loop sizes upon adsorption on the gold SPR sensor surface. Dodecyltrimethylammonium chloride in solution was flowed over the polymer-coated sensor surface and the binding was followed in real time. It was found that control of the loop size of the polymer on the solid surface enabled in turn the control of surfactant binding, with the largest loop allowing the maximum amount of surfactant to bind and vice versa. The kinetic plot of the binding showed three distinct segments. The first segment followed convective-diffusive kinetics. The second and third segments followed first-order kinetics with the second rate being significantly faster than the first one. Careful analysis of the second segment showed that it is possible to divide it into two different segments, each following a first-order kinetics, with the second rate being slightly slower than the first one suggesting a gradual slow down of the reaction due to convolution from the polymer conformational changes. Mechanistically, the sudden increase in the rate for the third segment of surfactant binding implies that the polymer matrix is opening up so as to incorporate more surfactant molecules. This was attributed to the formation of charged double surfactant species the repulsive interaction of which prevented the polymer network from imploding. Studies using unmodified polymers suggested the possibility of sudden conformational rearrangement in the polymer network, with progress in surfactant binding. Furthermore, the reflectance of the SPR spectrum was found to increase upon surfactant binding, implying that there is a decreased efficiency of coupling of the incident radiation into the surface plasmon mode of the metal, which suggests that the surfactant actually penetrated the polymer matrix. PMID- 12725840 TI - Effect of amino acid surfactants on phase transition of poly(N isopropylacrylamide) gel. AB - The volume phase transition behavior of a poly(N-isopropylacrylamide) gel (NIPA gel) in solutions of N-acyl amino acid surfactants were studied as a function of surfactant concentration. The addition of a surfactant beyond the critical micelle concentration (cmc) produced elevation in the transition temperature of the NIPA gel and its swelling. The changes in the volume phase transition temperature and in the swelling of the NIPA gel became more significant with the decreasing size of the amino acid side chain. This result could almost be explained only by the binding amount of surfactant onto the NIPA gel regardless of molecular structure of the amino acid. The binding amount increased in the order of sodium N-lauroyl-glycinate>-alaninate>-valinate>-leucinate>or= phenylalaninate. For an N-acyl amino acid surfactant to bind onto the NIPA gel, to increase the transition temperature, and to facilitate swelling of the gel, the steric hindrance of the amino acid side chain was more effective than its hydrophobicity. PMID- 12725842 TI - Parents' perceptions of their 1-6-year-old children's pain. AB - BACKGROUND OF THE STUDY: Parents' perceptions of children's pain may have influence on how children's postoperative pain is alleviated at home after discharge from hospital. PURPOSE OF THE STUDY: To describe parents' perceptions of 1-6-year-old children's pain. METHODS: Mothers (N=201) and fathers (N=114) whose child had undergone a day surgery in 10 Finnish central hospitals between October 2000 and September 2001 filled in a questionnaire including statements of pain perceptions, a Visual Analogue Scale to assess children's pain intensity and Parents' Postoperative Pain Measure (PPPM) to measure children's pain behaviours. RESULTS: Most of the parents suggested that adults have the responsibility to alleviate child's pain and that alleviation of child's postoperative pain prevents the child's fears during future visits in child welfare clinic. However, majority of parents described that postoperative pain decreases every day or that pain is always a part of surgery. Differences in parents' perceptions were found by both parents' and children's background variables. Parents' perceptions of children's pain were related to children's pain intensity and pain behaviours after surgery. CONCLUSIONS: Parents' perceptions of children's pain were related to children's pain after surgery at home. Adequate information of children's pain should be provided to the parents before discharge to promote children's pain alleviation at home. Special attention should be paid on parents' expectations of boys' higher pain tolerance. PMID- 12725843 TI - Colored pain drawings: preliminary observations in a neurosurgical practice. AB - BACKGROUND: Black and white pain drawings were introduced as a proposed means to identify patients, presenting with low back pain, who demonstrated functional overlay upon neurological testing. The use of color may enhance the usefulness of such pain drawings, but has not been described for adult patients. AIMS: To retrospectively explore the use of colored pain drawings in patients with neck, low back, or radicular pain. METHODS: Patients with neck, low back, or radicular pain referred to a community-based neurosurgical practice for evaluation during 1 year (n=359) depicted their pain on anatomical drawings using colored pencils representing different pain characteristics. Patients with abnormal (n=55) and normal (n=54) pain drawings were selected for this study. Use of medications, findings on physical examination, radiographic findings, activity levels, Waddell signs, and pending litigation were recorded and compared between patients with normal and abnormal pain drawings, as assessed according to the Ransford penalty point system. RESULTS: Patients whose colored pain drawings were abnormal, demonstrated a greater use of medications, more non-focal clinical findings, Waddell signs, impaired activity levels, involvement in pending litigation, and significantly fewer pathological radiographic findings than patients with normal pain drawings. CONCLUSIONS: Our findings agree with previous observations using black and white pain drawings, indicating that colored pain drawings are no less useful than the black and white approach. Further research is necessary to examine the psychometric properties and clinical usefulness of colored pain drawings to predict outcomes and/or determine treatment. PMID- 12725844 TI - Patient reporting of adverse drug reactions: useful information for pain management? AB - BACKGROUND: Patients' perceptions of adverse effects caused by the medicines they are prescribed may influence how they use these medicines. Little is known about patients' perceptions of the adverse effects of specific drugs in everyday use and whether these differ from those identified by clinical trials and standard post-marketing surveillance methods. AIM: To compare reports of perceived adverse drug reactions (ADRs) obtained directly from patients taking tramadol to those found in clinical trials and two methods of post-marketing surveillance. METHOD: Postal questionnaire distributed to 1048 patients who had a prescription for tramadol dispensed over a 3-month period. RESULTS: Most (84%) of the 344 respondents reported at least one symptom perceived as an ADR to tramadol. Dry mouth, light-headedness and constipation were most commonly reported. Almost half (48%) rated their most bothersome symptom as at least moderate and 43% claimed to have reported symptoms to their doctor. Perceived problems had led 38 respondents to stop taking tramadol. The 10 most frequently reported symptoms were all previously reported ADRs to tramadol. Although relatively minor, all 10 also appeared in reports to the UK Committee on the Safety of Medicines (CSM) and in prescription event monitoring. For many symptoms, the estimated range of frequency was in line with published reports, but considerably higher than that of post-marketing surveillance methods. CONCLUSIONS: Symptoms were reported by the majority of respondents and for many symptoms the frequency was high. Many patients did not report symptoms they perceived to be adverse effects to their doctor. The results indicate that patient perceptions of potential ADRs are relevant and should be an integral part of a pain management strategy. PMID- 12725845 TI - Long term depression of human nociceptive skin senses induced by thin fibre stimulation. AB - We have recently shown that stimulation, through a multi-electrode array, of thin nerve fibres close to the dermo-epidermal junction in the skin, produces powerful inhibition of itch and, to a lesser degree, cutaneous pain in humans. Here, we have studied the induction time and frequency dependency (range 1-10Hz) of the inhibitory effects of such stimulation on itch, mechanical, and thermal pain, in 20 healthy subjects. Sixteen electrodes applied on the skin were consecutively stimulated using a method termed cutaneous field stimulation (CFS). The results show that different treatment periods with CFS were required for the induction of significant inhibitory effects on different nociceptive qualities: 1st heat pain (1 min), itch (3 min), 2nd heat pain (6 min), pinch evoked pain (8 min). Six to ten minutes stimulation sufficed to induce peak inhibitory effects on all these sensory qualities while longer stimulation (up to 40 min) did not cause significantly stronger inhibition. The effects on itch, 1st and 2nd heat pain lasted over 55 min after termination of CFS. There was no effect on prickle. No significant difference in inhibitory effects of different stimulation frequencies (1, 4 and 10Hz/electrode) was found. The induction time and effective stimulation frequencies may suggest that the underlying mechanisms are similar to those of long term depression (LTD) previously described in the spinal cord in animal experiments. PMID- 12725846 TI - Pain as presenting symptom in Lyme neuroborreliosis. AB - Neurogenic pain with radiculitis is often the starting symptom in adult patients with tick-borne Lyme neuroborreliosis and in some cases the only clinical manifestation. Cranial paresis and other neurologic signs usually occur after the onset of pain. The present paper describes four patients who had severe pain as the main presenting symptom of Lyme neuroborreliosis. Opioids had good short-term effect in two of the cases. Oral doxycycline treatment was used successfully to eliminate the infection. PMID- 12725847 TI - Antinociceptive effects of RB101(S), a complete inhibitor of enkephalin catabolizing enzymes, are enhanced by (+)-HA966, a functional NMDA receptor antagonist: a c-Fos study in the rat spinal cord. AB - The effects of the S enantiomer of RB101, a complete inhibitor of enkephalin catabolizing enzymes, alone or in combination with a functional NMDA receptor antagonist, (+)-HA966 were studied on the spinal c-Fos protein expression in the carrageenan model of inflammatory nociception. One hour 30min after intraplantar carrageenan in awake rats, c-Fos immunoreactive (c-Fos-IR) nuclei were preferentially located in the laminae I-II and V-VI of the spinal dorsal horn, i.e., spinal areas containing numerous neurons responding exclusively, or not, to peripheral nociceptive stimuli. RB101(S) (5, 10, 20 and 40mg/kg i.v.) dose dependently reduced the total number of carrageenan-evoked c-Fos-IR nuclei (r=0.63, P<0.01), with 49+/-3% reduction (P<0.001) for the highest dose. Two highest doses of RB101(S) (20 and 40mg/kg) significantly reduced the number of carrageenan-evoked c-Fos-IR nuclei in both superficial I-II (32+/-7% and 36+/-5% reduction, respectively, P<0.05 for both) and deep V-VI (42+/-6% and 61+/-2% reduction, respectively, P<0.001 for both) laminae. The effects of RB101(S) were naloxone-reversible. Combination of low doses of RB101(S) (2.5 or 10mg/kg i.v.) and an inactive dose of (+)-HA966 (2.5mg/kg s.c.) produced supra-additive effects (39+/-4% and 51+/-5% reduction of the total number of c-Fos-IR nuclei, respectively, P<0.001 for both). These effects were partially reversed by naloxone. These results provide evidence for the potent effects of combination of RB101(S) and (+)-HA966. Considering the absence of major opioid side effects of RB101(S) and the marked increase of its antinociceptive effects by NMDA receptor antagonist, this type of drug combination could have beneficial therapeutical application. PMID- 12725848 TI - Segmental and plurisegmental modulation of pressure pain thresholds during static muscle contractions in healthy individuals. AB - The purpose of this study was to assess possible segmental (uni- and/or bilateral) and plurisegmental changes in pressure pain thresholds (PPTs) during static muscle contractions. Twenty-four healthy subjects (12 female, 12 male) performed a standardised isometric contraction with the dominant m. quadriceps femoris (MQF) and m. infraspinatus (MI), respectively. PPTs were assessed using pressure algometry at the contracting muscle, at the contralateral (resting) muscle and at a distant resting muscle (MI during contraction of MQF and vice versa). The PPT assessments were performed before, during and 30min. following each contraction. The contractions were held until exhaustion or for a maximum of 10 PPT assessments/muscle. During contraction of MQF PPTs increased compared to baseline at the middle ( p<0.001) and the end (p<0.001) of the contraction period at all assessed sites alike. During contraction of MI PPTs increased compared to baseline at the middle (p<0.001) and the end (p<0.007) of the contraction period at all sites. The increase was more pronounced at the contracting muscle compared to the contralateral (p<0.002; p<0.01) and the distant (p<0.002; p<0.002) site. No statistically significant difference was seen in PPTs between the latter two. Following the contractions PPTs returned to baseline. Submaximal isometric contraction of MQF and MI gave rise to a statistically significant increase in PPTs at the contracting muscle, the resting homologous contralateral muscle and at the distant resting muscle indicating that generalised pain inhibitory mechanisms were activated. Contraction of MI, but not of MQF, gave rise to an additional activation of unilateral segmental antinociceptive effects. PMID- 12725849 TI - Interobserver reliability of diagnosis in patients with complex regional pain syndrome. AB - Complex regional pain syndrome type I (CRPS-I), formerly reflex sympathetic dystrophy (RSD), is a chronic pain syndrome of unknown aetiology. Its diagnosis is a clinical one, for which several criteria systems have been defined. Despite their widespread use, the reliability of these criteria has never been studied. In this interobserver study 25 chronic CRPS patients were interviewed and examined by six physicians. Through structured questionnaires signs, symptoms, and diagnosis were recorded, after which observer agreement for these was calculated with kappa statistics. Physicians' agreement in assessment of signs and symptoms in CRPS patients varied greatly. More importantly, final diagnosis of CRPS showed poor observer agreement (kappa: 0.20). The kappa values were higher, had physicians applied IASP criteria, but still insufficient. The application of Bruehl's criteria results in a fair kappa of 0.38, but then frequency of CRPS diagnosis in our study population decreased from 73% to 43% in comparison with physicians' own diagnosis. We conclude that, using current criteria systems, the diagnosis of CRPS is not reliable. PMID- 12725850 TI - Somatosensory perception in patients suffering from long-term trapezius myalgia at the site overlying the most painful part of the muscle and in an area of pain referral. AB - In subgroups of patients with localised musculoskeletal pain spread of pain and signs of altered somatosensory processing at painful sites, both focal and referred areas have been reported. The purpose of the study was to examine somatosensory processing in patients with mainly unilateral long-term (> or =1 year) trapezius myalgia with ongoing pain for the last 3 months in the trapezius muscle in conjunction with ongoing or recurrent referral of pain to the ipsilateral arm. Ten patients with trapezius myalgia and 10 age- and sex-matched healthy controls participated. Pressure pain sensitivity, low threshold mechanoreceptive function and thermal sensitivity, including thermal pain, were assessed at the site overlying the most painful part of the trapezius muscle and in an area of pain referral in the ipsilateral upper arm/forearm as well as in the corresponding contralateral areas. No significant difference in sensibility was found in the most affected trapezius muscle and contralaterally compared to the corresponding areas in controls. In the area of pain referral there was a significantly increased sensitivity to pressure pain compared to the homologous contralateral area (p<0.01) as well as to the corresponding area in controls (p<0.009). Compared to controls a bilaterally decreased sensitivity to light touch was found in patients in the area of referred pain (p<0.01). No differences were found in the outcome of thermal testing. These findings suggest altered central processing of somatosensory input from the area of referred pain in patients with trapezius myalgia. PMID- 12725851 TI - Experimental deep tissue pain in wrist extensors--a model of lateral epicondylalgia. AB - The aim of this experimental study was to develop an in vivo model demonstrating sensory and motor interactions comparable to those seen in patients presenting with lateral epicondylalgia (i.e., deep tissue pain and hyperalgesia localised to specific sites in the wrist extensors, attenuation of wrist extension force). The effect of saline-induced deep pain combined with delayed onset muscle soreness (DOMS) on deep tissue sensitivity and motor function in wrist extensors was examined. Muscle pain intensity (visual analogue scale: VAS), distribution, and quality were assessed in 12 subjects. Pressure pain thresholds (PPTs) were recorded at five different sites around the elbow. Maximal wrist extension force was recorded. In the absence of DOMS, hypertonic saline administrated into different parts of the wrist extensors (extensor carpi radialis brevis, supinator, common extensor origin) induced significantly (P<0.05) higher VAS scores and larger pain areas compared with a control injection of isotonic saline. The typical quality of saline-induced pain was described as "drilling", "taut", "nagging" and "intense". In non-exercised wrist extensors, hyperalgesia to pressure was not detected during saline-induced pain but maximal wrist extensor force decreased significantly (P<0.05) compared with pre-pain recordings and recordings post isotonic saline. DOMS induced by eccentric wrist extension contractions generated moderate levels of soreness but no resting pain up to 24h post exercise. PPTs and maximal wrist extension force were significantly decreased (P<0.05) during DOMS compared with baseline and 7 days post exercise (P<0.05). VAS scores to injection of hypertonic saline into the DOMS arm were significantly increased (P<0.05) compared with injections into the unexercised arm. This is another manifestation of muscle hyperalgesia. Saline-induced pain combined with DOMS further decreased maximal wrist extension force (P<0.05). The simultaneous deep tissue pain and hyperalgesia linked with force attenuation support the use of the saline-induced deep tissue pain combined with DOMS as an experimental model simulating the clinical sensorimotor correlates of lateral epicondylalgia. PMID- 12725853 TI - Fibromyalgia may mask onset of autoimmune diseases. PMID- 12725852 TI - Treatment of chronic pain with millimetre wave therapy (MWT) in patients with diffuse connective tissue diseases: a pilot case series study. AB - BACKGROUND: Pain relief is reported to be the most common clinical application of electromagnetic millimetre waves. AIM: To evaluate safety and pain relief effect of millimetre wave therapy (MWT) for treatment of chronic joint pain in a group of patients with diffuse connective tissue diseases. METHODS: Twelve patients with diffuse connective tissue diseases received MWT in addition to their analgesic medication with non-steroidal anti-inflammatory drugs. MWT procedure included the exposure of tender points around the painful joints to electromagnetic waves with frequency 54-78GHz and power density of 2.5mW/cm(2). The time of exposure was 35 +/-5 min and the total number of sessions ranged from 5 to 10 (median 6). Intensity of pain, medication requirement, joint stiffness and subjective assessment of therapy success were measured before, during and immediately after the treatment, and after a 6-months follow-up. RESULTS: No adverse effects of MWT were noted. Pain intensity and required medication decreased significantly after the treatment (p<0.05) and remained at the same level throughout the follow-up period. The joint stiffness decreased and the subjective assessment of the treatment success after 6 month did not change except in only one patient. CONCLUSION: MWT applied to tender points around the affected joints was safe under the conditions of our study and after an appropriate full-scale double-blind clinical study, may be recommended as an effective adjunct therapy for chronic pain treatment in patients with diffuse connective tissue diseases. PMID- 12725854 TI - Surface Logix. Mini disease models for maximum impact. PMID- 12725855 TI - Infinity Pharmaceuticals. Back to the future. PMID- 12725856 TI - Protecting against cocaine, heroin, and sarin gas. AB - The first X-ray structure of human carboxylesterase 1 (hCE1) and the structures of hCE1 with drug analogs bound reveal important molecular details of how the drugs cocaine, heroin, and tacrine are metabolized and cleared. PMID- 12725858 TI - The biosynthetic gene cluster for the beta-lactam carbapenem thienamycin in Streptomyces cattleya. AB - beta-lactam ring formation in carbapenem and clavam biosynthesis proceeds through an alternative mechanism to the biosynthetic pathway of classic beta-lactam antibiotics. This involves the participation of a beta-lactam synthetase. Using available information from beta-lactam synthetases, we generated a probe for the isolation of the thienamycin cluster from Streptomyces cattleya. Genes homologous to carbapenem and clavulanic acid biosynthetic genes have been identified. They would participate in early steps of thienamycin biosynthesis leading to the formation of the beta-lactam ring. Other genes necessary for the biosynthesis of thienamycin have also been identified in the cluster (methyltransferases, cysteinyl transferases, oxidoreductases, hydroxylase, etc.) together with two regulatory genes, genes involved in exportation and/or resistance, and a quorum sensing system. Involvement of the cluster in thienamycin biosynthesis was demonstrated by insertional inactivation of several genes generating thienamycin nonproducing mutants. PMID- 12725859 TI - Directed evolution of O6-alkylguanine-DNA alkyltransferase for efficient labeling of fusion proteins with small molecules in vivo. AB - We report here the generation of mutants of the human O(6)-alkylguanine-DNA alkyltransferase (hAGT) for the efficient in vivo labeling of fusion proteins with synthetic reporter molecules. Libraries of hAGT were displayed on phage, and mutants capable of efficiently reacting with the inhibitor O(6)-benzylguanine were selected based on their ability to irreversibly transfer the benzyl group to a reactive cysteine residue. Using synthetic O(6)-benzylguanine derivatives, the selected mutant proteins allow for a highly efficient covalent labeling of hAGT fusion proteins in vivo and in vitro with small molecules and therefore should become important tools for studying protein function in living cells. In addition to various applications in proteomics, the selected mutants also yield insight into the interaction of the DNA repair protein hAGT with its inhibitor O(6) benzylguanine. PMID- 12725860 TI - Analysis of the importance of the metallo-beta-lactamase active site loop in substrate binding and catalysis. AB - The role of the mobile loop comprising residues 60-66 in metallo-beta-lactamases has been studied by site-directed mutagenesis, determination of kinetic parameters for six substrates and two inhibitors, pre-steady-state characterization of the interaction with chromogenic nitrocefin, and molecular modeling. The W64A mutation was performed in IMP-1 and BcII (after replacement of the BcII 60-66 peptide by that of IMP-1) and always resulted in increased K(i) and K(m) and decreased k(cat)/K(m) values, an effect reinforced by complete deletion of the loop. k(cat) values were, by contrast, much more diversely affected, indicating that the loop does not systematically favor the best relative positioning of substrate and enzyme catalytic groups. The hydrophobic nature of the ligand is also crucial to strong interactions with the loop, since imipenem was almost insensitive to loop modifications. PMID- 12725861 TI - Sexual attraction in the silkworm moth. Nature of binding of bombykol in pheromone binding protein--an ab initio study. AB - An analysis of the crystal structure of [BmPBP...bombykol] complex identified nine amino acid residues involved in a variety of intermolecular interactions binding the ligand. Using simple model fragments as the representatives of the residues, the interaction energies of their complexes with bombykol were calculated using high-level ab initio methods. The results were discussed in terms of the method and basis set dependence and were further corrected to account for their pair nonadditivities. This enabled us to describe quantitatively the nature and origin of the binding forces in terms of contribution of the individual amino acids and individual types of interaction to the overall stability. All of these interactions are well defined and cannot be considered as nonspecific hydrophobic interactions, one of the major conclusions of this work. PMID- 12725862 TI - Crystal structure of human carboxylesterase 1 complexed with the Alzheimer's drug tacrine: from binding promiscuity to selective inhibition. AB - Human carboxylesterase 1 (hCE1) is a broad-spectrum bioscavenger that plays important roles in narcotic metabolism, clinical prodrug activation, and the processing of fatty acid and cholesterol derivatives. We determined the 2.4 A crystal structure of hCE1 in complex with tacrine, the first drug approved for treating Alzheimer's disease, and compare this structure to the Torpedo californica acetylcholinesterase (AcChE)-tacrine complex. Tacrine binds in multiple orientations within the catalytic gorge of hCE1, while it stacks in the smaller AcChE active site between aromatic side chains. Our results show that hCE1's promiscuous action on distinct substrates is enhanced by its ability to interact with ligands in multiple orientations at once. Further, we use our structure to identify tacrine derivatives that act as low-micromolar inhibitors of hCE1 and may provide new avenues for treating narcotic abuse and cholesterol related diseases. PMID- 12725863 TI - Rational engineering of a DNA glycosylase specific for an unnatural cytosine:pyrene base pair. AB - A novel site-specific cytosine DNA glycosylase has been rationally engineered from the active site scaffold of the DNA repair enzyme uracil DNA glycosylase (UDG). UDG, which operates by a nucleotide flipping mechanism, was first converted into a sequence nonspecific cytosine DNA glycosylase (CDG) by altering the base-specific hydrogen bond donor-acceptor groups in the active site. A second mutation that renders UDG defective in nucleotide flipping was then introduced, and the double mutant was rescued using a substrate with a "preflipped" cytosine base. Substrate-assisted flipping was engineered by incorporation of an unnatural pyrene nucleotide wedge (Y) into the DNA strand opposite to the target cytosine. This new enzyme, CYDG, can be used to target cleavage of specific cytosine residues in the context of a C/Y base pair in any DNA fragment. PMID- 12725864 TI - Crystal structure and molecular modeling of 17-DMAG in complex with human Hsp90. AB - Hsp90 is an attractive chemotherapeutic target because it chaperones the folding of proteins found in multiple signal transduction pathways. We describe the 1.75 A resolution crystal structure of human Hsp90 alpha (residues 9-236) complexed with 17-desmethoxy-17-N,N-dimethylaminoethylamino-geldanamycin (17-DMAG). The structure revealed an altered set of interactions between the 17-substituent and the protein compared to geldanamycin and the 17-dimethylaminoethyl moiety pointing into solvent, but otherwise was similar to that reported for the complex with geldanamycin. Targeted molecular dynamics simulations and energetic analysis indicate that geldanamycin undergoes two major conformational changes when it binds Hsp90, with the key step of the conversion being the trans to cis conformational change of the macrocycle amide bond. We speculate that 17-DMAG analogs constrained to a cis-amide in the ground state could provide a significant increase in affinity for Hsp90. PMID- 12725867 TI - Conformation, action, and mechanism of action of neuromuscular blocking muscle relaxants. AB - Since curare was introduced into clinical anaesthesia in 1942, efforts to create better neuromuscular blocking (NMB) muscle relaxants have continued. Today, muscle relaxation remains a mainstay of modern anaesthesia and intensive care. Through manipulation of the traditional structure-action relationships, many new and improved muscle relaxants have been created, and several have been brought to clinical use. However, structure-action relationship is inconsistent and has its limits. Using computer-aided molecular conformational analyses, the conformation action relationships of NMB agents of various chemical classes have been explored. Conformation, no less than structure, of the NMB agents has shed new light on their mechanisms of action. By reflection, the conformations also suggest new details of the topology of the receptive sites of the nicotinic acetylcholine receptor modeled for the motor endplate of the skeletal muscle. PMID- 12725866 TI - The p38 mitogen-activated protein kinase pathway and its role in interferon signaling. AB - Interferons (IFNs) are pleiotropic cytokines that exhibit multiple biological effects on cells and tissues. IFN receptors are expressed widely in mammalian cells and virtually all different cell types express them on their surface. The Type I IFN receptor has a multichain structure, composed of at least two distinct receptor subunits, IFNalphaR1 and IFNalphaR2. Two Jak-kinases, Tyk-2 and Jak-1, associate with the different receptor subunits and are activated in response to IFNalpha or IFNbeta to regulate engagement of multiple downstream signaling cascades. These include the Stat-pathway, whose function is essential for transcriptional activation of IFN-sensitive genes, and the insulin receptor substrate pathway, which regulates downstream activation of the phosphatidyl inositol-3' kinase. Members of the Map family of kinases are also activated by the Type I IFN receptor and participate in the generation of IFN signals. The p38 Map kinase pathway appears to play a very important role in the induction of IFN responses. p38 is rapidly activated during engagement of the Type I IFN receptor, and such an activation is regulated by the small G-protein Rac1, which functions as its upstream effector in a tyrosine kinase-dependent manner. The activated form of p38 regulates downstream activation of other serine kinases, notably MapKapK-2 and MapKapK-3, indicating the existence of Type I IFN-dependent signaling cascades activated downstream of p38. Extensive studies have shown that p38 plays a critical role in Type I IFN-dependent transcriptional regulation, without modifying activation of the Stat-pathway. It is now well established that the function of p38 is essential for gene transcription via ISRE or GAS elements, but has no effects on the phosphorylation of Stat-proteins, the formation of Stat complexes, and their binding to the promoters of IFN-sensitive genes. As Type I IFNs regulate gene expression for proteins with antiviral properties, it is not surprising that pharmacological inhibition of the p38 pathway blocks induction of IFNalpha-antiviral responses. In addition, pharmacological inhibition of p38 abrogates the suppressive effects of Type I IFNs on normal human hematopoietic progenitors, indicating a critical role for this signaling cascade in the induction of the regulatory effects of Type I IFNs on hematopoiesis. p38 is also activated during IFNalpha-treatment of primary leukemia cells from patients with chronic myelogenous leukemia. Such activation is required for IFNalpha-dependent suppression of leukemic cell progenitor growth, indicating that this pathway plays a critical role in the induction of the antileukemic effects of IFNalpha. PMID- 12725868 TI - Neuroadaptive processes in GABAergic and glutamatergic systems in benzodiazepine dependence. AB - Knowledge of the neural mechanisms underlying the development of benzodiazepine (BZ) dependence remains incomplete. The gamma-aminobutyric acid (GABA(A)) receptor, being the main locus of BZ action, has been the main focus to date in studies performed to elucidate the neuroadaptive processes underlying BZ tolerance and withdrawal in preclinical studies. Despite this intensive effort, however, no clear consensus has been reached on the exact contribution of neuroadaptive processes at the level of the GABA(A) receptor to the development of BZ tolerance and withdrawal. It is likely that changes at the level of this receptor are inadequate in themselves as an explanation of these neuroadaptive processes and that neuroadaptations in other receptor systems are important in the development of BZ dependence. In particular, it has been hypothesised that as part of compensatory mechanisms to diazepam-induced chronic enhancement of GABAergic inhibition, excitatory mechanisms (including the glutamatergic system) become more sensitive [Behav. Pharmacol. 6 (1995) 425], conceivably contributing to BZ tolerance development and/or expression of withdrawal symptoms on cessation of treatment, including increased anxiety and seizure activity. Glutamate is a key candidate for changes in excitatory transmission mechanisms and BZ dependence, (1) since there are defined neuroanatomical relationships between glutamatergic and GABAergic neurons in the CNS and (2) because of the pivotal role of glutamatergic neurotransmission in mediating many forms of synaptic plasticity in the CNS, such as long-term potentiation and kindling events. Thus, it is highly possible that glutamatergic processes are also involved in the neuroadaptive processes in drug dependence, which can conceivably be considered as a form of synaptic plasticity. This review provides an overview of studies investigating changes in the GABAergic and glutamatergic systems in the brain associated with BZ dependence, with particular attention to the possible differential involvement of N-methyl-D-aspartate and alpha-amino-3-hydroxy-5 methyl-4-isoxazole propionic acid receptors in these processes. PMID- 12725869 TI - Regulation of muscarinic acetylcholine receptor signaling. AB - Multiple mechanisms regulate the signaling of the five members of the family of the guanine nucleotide binding protein (G protein)-coupled muscarinic acetylcholine (ACh) receptors (mAChRs). Following activation by classical or allosteric agonists, mAChRs can be phosphorylated by a variety of receptor kinases and second messenger-regulated kinases. The phosphorylated mAChR subtypes can interact with beta-arrestin and presumably other adaptor proteins as well. As a result, the various mAChR signaling pathways may be differentially altered, leading to short-term or long-term desensitization of a particular signaling pathway, receptor-mediated activation of the mitogen-activated protein kinase pathway downstream of mAChR phosphorylation, as well as long-term potentiation of mAChR-mediated phospholipase C stimulation. Agonist activation of mAChRs may also induce receptor internalization and down-regulation, which proceed in a highly regulated manner, depending on receptor subtype and cell type. In this review, our current understanding of the complex regulatory processes that underlie signaling of mAChR is summarized. PMID- 12725870 TI - Organization of multiple cytochrome P450s with NADPH-cytochrome P450 reductase in membranes. AB - Microsomal P450-mediated monooxygenase activity supported by NADPH requires an interaction between flavoprotein NADPH-cytochrome P450 reductase and cytochrome P450. These proteins have been identified as the simplest system (with the inclusion of a phospholipid (PL) component) that possesses monooxygenase function; however, little is known about the organization of these proteins in the microsomal membrane. Although reductase and P450 are known to form a 1:1 functional complex, there exists a 10- to 20-fold excess of P450 over the reductase. This raises several questions including "How are the enzymes of the P450 system organized in the microsomal membrane?" and "Can one P450 enzyme affect the functional characteristics of another P450?" This review summarizes evidence supporting the potential for enzymes involved in the P450 system to interact, focusing on the interactions between reductase and P450 and interactions between multiple P450 enzymes. Studies on the aggregation characteristics of P450 as well as on rotational diffusion are detailed, with a special emphasis on the potential for P450 enzymes to produce oligomeric complexes and to suggest the environment in which P450 exists in the endoplasmic reticulum. Finally, more recent studies describing the potential for multiple P450s to exist as complexes and their effect on P450 function are presented, including studies using reconstituted systems as well as systems where two P450s are coexpressed in the presence of reductase. An understanding of the interactions among reductase and multiple P450s is important for predicting conditions where the drug disposition may be altered by the direct effects of P450-P450 complex formation. Furthermore, the potential for one P450 enzyme to affect the behavior of another P450 may be extremely important for drug screening and development, requiring metabolic screening of a drug with reconstituted systems containing multiple P450s rather than simpler systems containing only a single form. PMID- 12725871 TI - Recent advances in alpha1-adrenoceptor pharmacology. AB - alpha(1)-Adrenergic receptors (ARs) mediate some of the main actions of the natural catecholamines, adrenaline and noradrenaline. They participate in many essential physiological processes, such as sympathetic neurotransmission, modulation of hepatic metabolism, control of vascular tone, cardiac contraction, and the regulation of smooth muscle activity in the genitourinary system. Here, we review recent progress on subtype-specific subcellular localization, participation in signaling cascades, and the pivotal function of alpha(1)-ARs, as delineated through studies on genetically engineered animals. Together, these findings will provide new insights into the physiological and pathophysiological roles of the alpha(1)-ARs. PMID- 12725873 TI - Genealogy, expression, and cellular function of transforming growth factor-beta. AB - The transforming growth factor-beta (TGF-beta) gene superfamily expresses a large set of structurally and functionally related polypeptides. Three TGF-beta isoforms are regulated by specific genes and have been identified in mammals (TGF beta1, -beta2, and -beta3). All three-protein isoforms are observed abundantly during development and display overlapping and distinct spatial and temporal patterns of expressions. Each isoform plays a distinct role, the nature of which depends on the cell type, its state of differentiation, and growth conditions, and on the other growth factors present. TGF-beta regulates many of the processes common to both tissue repair and disease, including angiogenesis, chemotoxins, fibroblast proliferation and the controlled synthesis, and degradation of matrix proteins, such as collagen and fibronectin. This review will examine the genealogy and mode of actions of TGF-beta on the cell types involved in inflammation and repair, as well as in carcinoma. PMID- 12725872 TI - Glatiramer acetate (Copaxone) therapy for multiple sclerosis. AB - Glatiramer acetate (GA) (Copaxone(R)) is a worldwide-approved drug for the treatment of relapsing multiple sclerosis (MS), an autoimmune disease of the CNS. The drug is a synthetic copolymer with an amino acid composition based on the structure of myelin basic protein, one of the autoantigens implicated in the pathogenesis of MS and experimental autoimmune encephalomyelitis (EAE). Developed initially as a "tool" to study EAE, the drug unexpectedly inhibited disease and was subsequently developed for the treatment of MS. The drug has been shown in controlled clinical trials to significantly reduce relapse rate and progression of disability in MS with long-term efficacy, remarkable safety, and tolerability. Efficacy as measured by magnetic resonance imaging parallels its clinical benefits as manifested by a reduction in gadolinium-enhancing lesions and brain atrophy. The mechanism of action of the drug in humans is believed to involve the induction of glatiramer-reactive regulatory cells, including CD4+ and CD8+ T cells. Glatiramer-reactive Th2 cells are believed to enter the brain and, through cross-reactivity with myelin antigens, produce bystander suppression, antiinflammatory effects, and neuroprotection. PMID- 12725874 TI - Interobserver agreement in the histologic diagnosis of colorectal polyps. the experience of the multicenter adenoma colorectal study (SMAC). AB - Current clinical practice guidelines for patients with colorectal polyps are mainly based on the histologic characteristics of their lesions. However, interobserver variability in the assessment of specific polyp characteristics was evaluated in very few studies. The purpose of this study was to evaluate the interobserver agreement of four pathologists in the diagnosis of histologic type of colorectal polyps and in the degree of dysplasia and of infiltrating carcinoma in adenomas. A stratified random sample of 100 polyps was obtained from the 4,889 polyps resected within the Multicentre Adenoma Colorectal Study (SMAC), and the slides were blindly reviewed by the four pathologists. Agreement was analyzed using kappa statistics. A median kappa of 0.89 (range 0.79-1.0) was estimated for the interobserver agreement for the diagnosis of hyperplastic polyp vs. adenoma. The agreement in the diagnosis of tubular, tubulovillous, and villous type, was given by median kappa values of 0.50, 0.15, and 0.36, respectively. The median kappa for the diagnosis of infiltrating carcinoma was 0.78 (range 0.73-0.84). Agreement on diagnosis of adenoma histologic subtypes, degrees of dysplasia, or infiltrating carcinoma in adenoma was moderate. A simpler classifications might help to better identify patients at different risk of colorectal cancer. PMID- 12725876 TI - How to measure comorbidity. a critical review of available methods. AB - The object of this article was to systematically review available methods to measure comorbidity and to assess their validity and reliability. A search was made in Medline and Embase, with the keywords comorbidity and multi-morbidity, to identify articles in which a method to measure comorbidity was described. The references of these articles were also checked, and using a standardized checklist the relevant data were extracted from these articles. An assessment was made of the content, concurrent, predictive and construct validity, and the reliability. Thirteen different methods to measure comorbidity were identified: one disease count and 12 indexes. Data on content and predictive validity were available for all measures, while data on construct validity were available for nine methods, data on concurrent validity, and interrater reliability for eight methods, and data on intrarater reliability for three methods. The Charlson Index is the most extensively studied comorbidity index for predicting mortality. The Cumulative Illness Rating Scale (CIRS) addresses all relevant body systems without using specific diagnoses. The Index of Coexisting Disease (ICED) has a two-dimensional structure, measuring disease severity and disability, which can be useful when mortality and disability are the outcomes of interest. The Kaplan Index was specifically developed for use in diabetes research. The Charlson Index, the CIRS, the ICED and the Kaplan Index are valid and reliable methods to measure comorbidity that can be used in clinical research. For the other indexes, insufficient data on the clinimetric properties are available. PMID- 12725875 TI - The intra- and interobserver variability of ankle-arm blood pressure index according to its mode of calculation. AB - To propose a standardization of calculus of the ankle-arm index as a diagnostic tool in the clinical setting and epidemiology of peripheral arterial disease, we aimed to study the reproducibility of its measurement through 15 different modes of calculation. The study was performed in a group of 194 vascular laboratory outpatients of a tertiary center. The intra- and interobserver variability was assessed by the intraclass correlation coefficient of agreement and the Bland & Altman method. Methods where the numerator was calculated by the average of posterior tibial and dorsalis pedis artery systolic pressures revealed to be the best reproducible. According to this study and former researches on this topic, we recommend the use of the average of posterior tibial and dorsalis pedis artery systolic pressures of the weakest limb for the numerator and the average of systolic pressures of humeral arteries for the denominator of the ankle-arm index. PMID- 12725877 TI - Optimal matching with a variable number of controls vs. a fixed number of controls for a cohort study. trade-offs. AB - Matching is used to control for imbalances between groups, but the preferable strategy for matching is not always clear. We sought to compare two algorithms optimal matching with a fixed number of controls (OMFC), and optimal matching with a variable number of controls (OMVC). We compared the degree of bias reduction and relative precision using Monte Carlo simulations. We systematically changed the magnitude of the matching variable difference, the variance ratios of the matching variable in the exposed and unexposed groups, the sample size, and the number of unexposed subjects available for matching. OMVC always produced larger removal of bias than the OMFC. The mean percentage reduction of bias was 38.3 with the OMFC and 52.6 with OMVC. OMVC increased the variance 6%. OMVC should be employed when researchers have access to a pool of unexposed subjects because it removes more bias with little loss in precision. PMID- 12725878 TI - Asthma-like symptoms assessment through ECRHS screening questionnaire scoring. AB - The definition of asthma has always been a matter of discussion. The European Community Respiratory Health Survey (ECRHS) is a multinational survey designed to compare the prevalence of asthma in subjects aged 20 to 44 years throughout European countries. In each center a representative sample completed a self administered screening questionnaire composed of nine dichotomous items. We propose a method for constructing a score for the ECRHS screening questionnaire and we validate the method with reference to the clinical diagnosis available for the Italian centers. Clinical diagnosis was made by a group of medical experts after examining the responses to a standardized clinical interview, respiratory function tests, and allergy tests. Before constructing a summary score, the number of latent factors/dimensions explaining correlations among the observed items was recognized. We identified only one factor/dimension underlying the screening questionnaire, so a summary score was determined by the Homogeneity Analysis by Alternating Least Square (HOMALS). Using best Receiver Operating Characteristic (ROC) threshold the diagnostic test of the score had sensitivity and specificity of 75.1 and 80.1%, respectively. The method of scoring is easily reproducible, and has the advantage of optimizing information recoded by the questionnaire. PMID- 12725879 TI - Meaningful effect size and patterns of response of the transition dyspnea index. AB - Ths object of this study was to examine validity, meaningful effect sizes, and patterns of response of the Transition Dyspnea Index (TDI) in a clinical trial cohort of chronic obstructive pulmonary disease (COPD) patients. The design was a retrospective analysis of data from a randomized, double-blind placebo-controlled clinical trial. We analyzed fifty clinical investigation sites in United States. There were 921 patients with stable COPD. Tiotropium 18 microg dry powder or matching placebo was used. Patients were allowed to remain on usual care less ipratropium bromide. Construct validity was demonstrated by significant correlations (P <.05) between Baseline Dyspnea Index (BDI) and other baseline measures, as well as between TDI and changes in other measures at the end of 1 year. Concurrent validity was observed by the significant correlation between TDI and dyspnea diary responses. Changes in TDI focal score were in the range of one unit when the group was stratified by a minimal change in the physician's global evaluation. Significantly less (P <.05) supplemental albuterol was observed in the group of responders defined by a one-unit improvement in TDI. Responders also had few exacerbations and better health status. The validity of the TDI is supported in a large clinical trial setting. A one-unit change in the TDI focal score represented the minimal important difference. PMID- 12725880 TI - Irritable bowel syndrome: are incentives useful for improving survey response rates? AB - Irritable bowel syndrome (IBS) is one of the most common gastrointestinal disorders. There continues to be a need for community-based research into this condition. Unfortunately, response rates in community-based IBS surveys have typically been very low. In this study, we explore the use of incentives and multiple-response options as a means of increasing survey response rates. The study was conducted in three phases. In an initial phase, no incentive was offered; in the second phase, a 5.00 Canadian dollars incentive was offered; and in the third phase, a 20.00 Canadian dollars incentive was offered. Response rates were higher in the incentive groups: Individual response rates were 57.9%, 72.7%, and 84.7% in the three phases, respectively. A slightly higher estimate of IBS prevalence was obtained in the no incentive group. Selection bias is a possible explanation for this difference. A decision about whether to use incentives must be based on the specific goals of the study. PMID- 12725881 TI - Stroke subtype and mortality. a follow-up study in 998 patients with a first cerebral infarct. AB - The aim of this article was to study mortality following a first-ever cerebral infarct, accounting for ischemic stroke subtypes (lacunar, cardioembolic, atherothrombotic) and relevant prognostic variables. This study was done from s a hospital-based prospective registry of all patients with a first cerebral infarct, with a high case ascertainment of first and recurrent stroke by CT. We used a cross-sectional follow-up, using standardized methods. Analyses were performed using crude comparison of mortality data and death causes between stroke subtypes. We analyzed 30-day case fatality and 1-year mortality in 30-day survivors by means of logistic regression analysis, and mortality in 1-year survivors by means of Cox proportional hazard modeling. We also constructed Kaplan-Meier survival curves, and used log-rank testing for differences between stroke subtypes. Thirty-day case fatality was 10%, 1-year mortality 15%, and after 1-year mortality 16%. Mean follow-up was 691, SD 521 days. At the end of follow-up 36% of all patients had died. Mortality was at all three time points lowest in lacunar stroke (2, 12, and 14%, respectively), intermediate in atherothrombotic stroke (10, 16, and 15%, respectively), and highest in cardioembolic stroke (23, 22, and 21%, respectively). Death related to recurrent stroke was similar in all three stroke subtypes (13-16%). Although 30-day case fatality rate was low in lacunar stroke, a quarter of lacunar stroke patients had died at the end of follow-up. Diabetes mellitus, age, stroke subtype, and initial stroke severity were independent predictors of 30-day case fatality, but only diabetes and age were consistent independent predictors for later mortality. Recurrent stroke and heart failure were important death causes. Prognosis for (future) death following a first cerebral infarct differs between stroke subtypes; lacunar stroke patients have the lowest mortality. However, lacunar stroke cannot be regarded as a mild stroke type, as after 2 years more than a quarter of such stroke patients had died. Cardioembolic stroke patients have the grimmest prognosis: more than half of them had died within 1.5 years. Better prognosis for long-term survival following stroke may be achieved by therapies which lower the risk of stroke recurrence, provide better treatment of heart failure, or both. PMID- 12725883 TI - No inverse association between fish consumption and risk of death from all causes, and incidence of coronary heart disease in middle-aged, Danish adults. AB - This study investigate the relation between fish consumption, all-cause mortality, and incidence of coronary heart disease (CHD). A total of 4,513 men and 3,984 women aged 30-70 years, sampled randomly from the population in Copenhagen County, Denmark, with initially examination in 1982-1992 was followed until 2000 for all-cause mortality and until 1997 for first admission to hospital or death from CHD. Information on fish consumption was obtained from a self administered food-frequency questionnaire. Cox proportional hazard analysis gave no evidence for an inverse association between fish consumption and all-cause mortality or incident CHD after adjustment for confounders. Among subjects with a priory-defined high risk of CHD there was a nonsignificant inverse relation between fish intake and CHD morbidity (Hazard Ratio 1.28 (0.92-1.80) for a consumption of fish of less than two times per month or less compared with once a week), but there was relatively few cases in this subgroup. These data provides no evidence for a protective effect of fish consumption on all-cause mortality or incident CHD in the population as a whole, but it cannot be excluded that frequent consumption of fish benefits those at high risk for CHD. PMID- 12725882 TI - Small area variation in childhood diabetes mellitus in Austria: links to population density, 1989 to 1999. AB - We analyzed the associations of environmental factors with the regional distribution of Type 1 diabetes mellitus in Austria. All newly diagnosed cases (n=1449) from 1989 to 1999 were allocated to districts using the postal code. Nitrate content of the water was measured by the Austrian Federal Environmental Agency. Data on infant mortality, population density, and percentage of employment by industry were derived from Statistics Austria. An inverse effect was seen between the proportion of children younger than 15 years of age and the risk ratio (P<.01). Infant mortality, population density, and percentage of persons with employment in industry were not of significant influence. The mean nitrate level was positively associated (P=.07). In regions with a higher percentage of children younger than 15 years of age, fewer children developed diabetes, which is in agreement with the observation that early social mixing is a protective factor. Nitrate levels may have a confounding effect. PMID- 12725884 TI - Statin use and the risk of breast cancer. AB - The study objective was to investigate a possible association between statin use and breast cancer (BRCA). An historical cohort design based on Saskatchewan's population health services databases was used. All eligible women with > or = 1 statin prescription from 1989 to mid-1997 and an age-sex-matched nonexposed group were followed up to 8.5 years (mean 4.2 years). Relative rates (RR) of BRCA were estimated and stratified by age, statin exposure time, and prior hormone use. Thirteen thousand five hundred ninety-two statin users and 53,880 nonexposed subjects were identified. Eight hundred seventy-nine incident BRCA cases were identified. Statins were not associated with BRCA risk in women < or = 55 years. Among subjects >55 years, the RR for BRCA was 1.15 (0.97, 1.37). Stratified analyses revealed increases in risk in short-term statin users and statin users with long-term hormone replacement therapy (HRT) exposure. More studies are needed to determine if short-term statin use and statin use with long-term HRT exposure increases postmenopausal BRCA risk. Published by Elsevier Science Inc. PMID- 12725885 TI - Is the APNCU Index to study LBW biased or only the article by Koroukian and Rimm? PMID- 12725887 TI - Apportionment of disease in individuals. PMID- 12725889 TI - Reconstructive neurosurgery for Parkinson's disease: a systematic review and preliminary meta-analysis. AB - This systematic review and meta-analysis aimed to identify the determinants for best practice and establish current benchmarks for recovery following reconstructive neurosurgery for people with Parkinson's disease. Eleven studies reporting results for 95 grafted patients were selected on the grounds of using optimal surgical techniques and the Core Assessment Program for Intracerebral Transplantation (CAPIT) protocol for data collection. Consistent trends demonstrating high levels of recovery were identified on most outcome measures. Determinants for best practice were identified as selecting younger patients; using low dose immunosuppression; bilateral grafting; and employing strategies to ensure the quantity and viability of the grafted cells. Secondary analysis of data demonstrated a correlation of rho=0.666 (P<0.05) between increases in striatal dopaminergic activity and UPDRS Motor (off) scores. Overall effect size 'd' was found to be 1.129 UPDRS Motor (off) condition and 0.719 for UPDRS Total (off) condition. The design of the studies and the variable standards for reporting the data precluded the use of more powerful and accurate meta-analyses. It was recommended that the creation of a collaborative database would improve the extraction of data and allow for more powerful statistical analyses for evaluating the overall harm and benefits associated with reconstructive neurosurgery. PMID- 12725890 TI - Topology and graph theory applied to cortical anatomy may help explain working memory capacity for three or four simultaneous items. AB - Cognitive experimentation suggests that at any single instant only three or four items ("chunks") are simultaneously prominent as a working memory (WM) trace, if we disregard the rehearsal component of WM. The reason for small WM capacity may concern combinatorial manageability. How might the neural representations of these few coactive chunks occupy a spatially distributed set of areas of the sheet-like cortex, while providing both order and flexibility to associate items in WM? Each attribute of each simultaneously active WM item must have broad access to the representational facilities of the cortical sheet, comprising tens of thousands of modular "cortical columns." The two hypothesized neural levels of WM during any moment of cognition comprise (a) "binding" together of many distributed attribute representations within each respective WM chunk, and (b) combinatorial play among three or four WM chunk-representations. Anatomical and functional evidence of cortical unity through its depth suggests that cortex may be viewed as essentially planar in its distribution of activations. Thus, a moment's WM is hypothesized here to reside in myriad activated cortical planar "patches," each subdivided into up to four amoeboid "subpatches." Two different lines of topological reasoning suggest orderly associations of such representations. (1) The four-color principle of map topology, and the related K(4) is planar theorem of graph theory, imply that if a small cortical area is dynamically subdivided into no more than four, discretely bounded planar subareas, then each such segment has ample free access to each of the others. (2) A hypothetical alternative to such associative adjacency of simultaneously active cortical representations of chunk-attributes is associative overlap, whereby, in dense cortical neuropil, activated subpatches behave like Venn diagrams of intersecting sets. As the number of Venn-like coactive subpatches within a patch increases, maintaining ad hoc associativity among all combinations requires exponentially proliferating intersections. Beyond four, serpentine subpatch shapes are required, which could easily lead to pathologies of omission or commission. As hypothesized by many researchers, the binding of the widely distributed cortical modules that represent a given chunk may involve synchrony or coherence of a single EEG frequency. Elsewhere, I have conjectured that such a binding frequency for a single chunk may bear a harmonic relationship with the additional EEG frequencies that are simultaneously binding the other WM chunks. Other possible mechanisms of binding have also been hypothesized. Whatever the mechanism, the many attributes of a moment's complement of three or four WM chunks must generally have an accidental relationship with the spatial distribution of the cortical feature analyzers that must be activated to represent those attributes. Therefore, the cortex may need, and have, comprehensive anatomical connections of each of its modules for representing an attribute (or of small redundant module groupings) with every other. If such whole-part cortico-cortical connections are somehow exploited not only to fully represent each cognitive chunk in its bound-together attributes, but also to bring the major business of intensive WM information processing down to the level of local circuits, in the sorts of topological patterning hypothesized here, there may be two adaptive results: (1) Time and other economies would be achieved in the reduction of activity in distant cortico-cortical connections to lower energy global orchestration, or binding processes. (2) The piecemeal local topological limit to four subpatches would be writ large, across the entire cortex, preventing an unconstrained combinatorial explosion of associations among all attributes of all three or four simultaneously active chunks. Such hypothetical convergence to foci in local subpatch interactions might take place primarily in association cortex, and/or it might involve temporary shifts in response properties in some cortical subpat might involve temporary shifts in response properties in some cortical subpatches. Quantitative studies of the densely packed cortical fine structure, by Braitenberg and Schuz, and others, seem potentially consistent with this vision of cortical function in cognition. PMID- 12725891 TI - On the confirmation of an effect of magnetic fields on the interictal firing rate of epileptic patients. AB - The effect of magnetic fields on interictal firing rates was investigated in three epileptic patients with depth electrode implantation in the hippocampus for pre-surgical evaluation. The protocol consisted of 10 min test periods, during which magnetic fields were cycled for 1 min on and 1 min off, and intervening 5 min rest periods. Only one patient revealed a 95% significant increase in the 10 s after the fields were switched on compared with the background estimate from the 10s before the fields were applied. This patient was also the only patient to show significant increases in firing rates during field-on compared with field off periods, and during magnetic field test periods compared with intervening rest periods. This patient had a right hippocampal seizure onset. All patients showed increased firing rates during the 10 min periods of magnetic field testing compared to the 5 min rest periods between tests. This result was significant for the group at the 99% level. Two patients with right temporal lobe onset showed greater activity in the right hippocampus than the left. All patients exhibited a progressive increase in firing rates in rest periods between tests. PMID- 12725892 TI - Short-term cerebral ischemia causes the dysfunction of interneurons and more excitation of pyramidal neurons in rats. AB - Neural excitotoxicity is a typical factor in the early phase pathogenesis of cerebral ischemia. Its cellular and molecular mechanisms are still unclear and clinical approaches are still lacking of promising therapies. We have examined the vulnerability of cortical neurons to short-term ischemia in rats by simultaneously analyzing the activities of inhibitory and principal neurons in brain slices. Our results demonstrate that short-term in vitro ischemia permanently impairs the excitability of inhibitory neurons (IN) and synaptic transmission mediated by gamma-aminobutyric acid (GABA). However, principal neurons appear to be more exciting during the reperfusion. The vulnerability of inhibitory neurons to ischemia acquires during postnatal development. Our findings signify a major contribution of the ischemic dysfunction of inhibitory neurons to neural excitotoxicity as well as a strategy to prevent the progress of ischemic stroke by protecting inhibitory neurons. PMID- 12725893 TI - A re-evaluation of the role of tetrahydropapaveroline in ethanol consumption in rats. AB - The role of tetrahydropapaveroline (THP), a condensation product of a dopaldehyde with dopamine, in the regulation of alcohol consumption was investigated. In the first experiment, rats received intraventricular injections of either racemic THP hydrobromide (0.65 or 1.3 microg/microl), R-(+)-THP (0.66 or 1.4 microg/microl), or an equal volume of vehicle. The lower doses of both (+/-)-THP and (+)-THP significantly increased volitional alcohol intake. For the racemic compound, the increase was significant at 7-13% concentrations. The R-(+)-enantiomer increased consumption at 4-11 and 15-20% concentrations of ethanol. The higher doses of both compounds did not significantly alter alcohol preference. A second experiment evaluated the chronic effect of THP delivered subcutaneously via osmotic minipump. Animals receiving THP (0.1, 0.5, 1.0, 2.0, and 4.0 mg/ml) did not differ in their alcohol intake, compared to vehicle-treated controls. Whether or not endogenously formed THP participates in the etiology of alcohol addiction remains unclear. Nonetheless, there are few known compounds that induce a preference for unsweetened alcohol solutions over water in laboratory animals. PMID- 12725894 TI - Metabotropic glutamate receptor-mediated hetero-synaptic interaction of red- and green-cone inputs to LHC of carp retina. AB - The role of presynaptic metabotropic glutamate receptor (mGluR) on the interaction of red- and green-cone signals was investigated in luminosity-type horizontal cell (LHC) of isolated carp retina. It was found that a dim red background could enhance LHC's light response to green stimulus, and a dim green background was also able to increase the cell's response to red flash. Such mutual color enhancement was eliminated by application of groups II and III mGluR antagonist (S)-methyl-4-carboxyphenyl-glycine (MCPG). Furthermore, inhibition of glutamate uptake by using D-aspartate (D-Asp) or DL-threo-beta-hydroxy-aspartic acid (THA) completely blocked the mutual enhancement of color signals in LHC. However, the GABAergic feedback pathway in the outer retina was unnecessarily involved. PMID- 12725895 TI - Synaptic facilitation by ectopic octopamine and 5-HT receptors in Aplysia. AB - The cAMP pathway plays a critical role in synaptic plasticity. We assessed using the ectopic expression of octopamine (OA) receptor, the contribution of the cAMP pathway to short-term facilitation of sensory-motor synapses in Aplysia. When synaptic connections were depressed to 20-30% of their initial EPSP amplitude, the application of OA to sensory cells expressing OA receptor showed significant synaptic facilitation, but this was less than the synaptic facilitation shown by 5-HT treatment. We also found that synaptic facilitation was further enhanced when OA was treated in the presence of 5-HT at non-depressed synapses, but not at depressed synapses. These results imply that the role of cAMP in synaptic facilitation is reduced as the synapse becomes depressed due to repeated activity. PMID- 12725896 TI - Participation of AMPA- and NMDA-type excitatory amino acid receptors in the spinal reflex transmission, in rat. AB - Classical in vitro and in vivo models and electrophysiological techniques were used to investigate the role of AMPA- and NMDA-type glutamate receptors in various components of spinal segmental reflex potentials. In the rat hemisected spinal cord preparation, the AMPA antagonists NBQX and GYKI 52466 abolished the monosynaptic reflex (MSR) potential but caused only partial inhibition of the motoneuronal population EPSP. NMDA antagonists had no noticeable effect on the MSR in normal medium, but markedly depressed the late part of EPSP. However, an NMDA receptor antagonist sensitive monosynaptic response was recorded in magnesium-free medium at complete blockade of the AMPA receptors. In spinalized rats, the AMPA antagonists completely blocked all components of the dorsal root stimulation evoked potential. MK-801 (2mg/kg, i.v.) reduced monosynaptic responses in a frequency dependent way, with no effect at 0.03 Hz and 22% inhibition at 0.25 Hz. The reduction of the di- and polysynaptic reflex components was about 30% and did not depend on stimulation frequency. Long latency reflex discharge responses, especially when evoked by train stimulation, were more sensitive to MK-801 than the polysynaptic reflex. These results suggest that glutamate activates MSR pathways through AMPA receptors. However, under certain conditions, NMDA receptors can modulate this transmission through plastic changes in the underlying neuronal circuits. AMPA and NMDA receptors play comparable roles in the mediation of longer latency reflex components. PMID- 12725897 TI - Microglial ecto-Ca(2+)-ATPase activity in a rat model of focal homologous blood clot embolic cerebral ischemia: an enzyme histochemical study. AB - Post-ischemic changes in ecto-Ca(2+)-ATPase activity in microglia and the infarcted tissue were studied in a rat model of focal embolic cerebral ischemia using an enzyme histochemical method. Ecto-Ca(2+)-ATPase activity was observed in whole brains in non-operated and sham-operated control animals. In addition, this enzyme activity was determined to be localized in ramified microglia. At 30 min after ischemia, non-microglial ecto-Ca(2+)-ATPase activity in the infarcted tissue slightly decreased and continued to decrease thereafter. The ecto-Ca(2+) ATPase activity in microglia did not appear changed at this time. The decrease of enzyme activity in the infarcted tissue made it much easier to clearly observe ecto-Ca(2+)-ATPase-positive microglia. The enzyme activity of microglia in the ischemic area began to decrease 2 or 4h after embolization and remarkably decreased, except in the perinuclear cytoplasm, apical parts of the processes, and several parts along the processes, 8h after ischemia. By 12h after onset of embolization, the enzyme activity of microglia and infarcted tissue had almost completely disappeared. Ecto-Ca(2+)-ATPase of microglia is likely to play an important role in the metabolism of extracellular nucleotides in the ischemic area immediately after the onset of embolization by means of ecto-enzymes. Thus, the findings of the present study suggest that microglia might serve to protect the infarcted tissue in the ischemic brain. PMID- 12725898 TI - The behavioral effects of magnesium therapy on recovery of function following bilateral anterior medial cortex lesions in the rat. AB - Magnesium (Mg(++)) therapy has been shown to be neuroprotective and to facilitate recovery of motor and sensorimotor function in a variety of animal models of traumatic brain injury. However, few studies have investigated the efficacy of Mg(++) therapy on cognitive impairments following injury. The present study evaluated the ability of magnesium chloride (MgCl(2)) to facilitate recovery of function following bilateral anterior medial cortex lesions (bAMC). Rats received electrolytic bAMC lesions or sham surgery and were then treated with 1 mmol/kg, i.p. MgCl(2), 2 mmol/kg, i.p. MgCl(2), or 1.0 ml/kg, i.p. 0.9% saline. Drug treatment was administered 15 min following injury with subsequent injections administered at 24 and 72 h. Rats were tested on a battery of behavioral tests that measured both cognitive (reference and working memory in the Morris Water Maze (MWM) and spatial delayed matching-to-sample (DMTS)) and sensorimotor performance (bilateral tactile adhesive removal). The results indicated that bAMC lesions produced significant cognitive impairments in reference memory and working memory in the MWM, DMTS and sensorimotor impairments compared to shams. Mg(++) therapy exhibited a dose-dependent effect in facilitating recovery of function. Administration of 2mmol of MgCl(2) significantly improved performance on the bilateral adhesive tactile removal test, DMTS and working memory tests. The 1 mmol dose of MgCl(2) reduced the initial deficit on the tactile adhesive removal test and reduced the working memory impairment on the second day of testing. These results suggest Mg(++) therapy improves cognitive performance following injury in a dose-dependent manner. PMID- 12725899 TI - Antidyskinetic effects of risperidone on animal models of tardive dyskinesia in mice. AB - The effects of risperidone, an atypical neuroleptic, were investigated on two animal models of tardive dyskinesia (TD). The repeated administration of reserpine (1.0mg/kg) or haloperidol (2.0mg/kg) induces orofacial movements in mice, which are very similar to those observed in humans presenting TD. The effects of acute or repeated treatment with several doses of risperidone (0.1; 0.5; 2.0 or 4.0) on the expression and development of orofacial movements in reserpine- and haloperidol-treated male mice were investigated. The results showed that risperidone per se did not induce the development of orofacial movements. In addition, this drug was able to attenuate the expression and the development of reserpine-as well as haloperidol-induced orofacial movements. These results are in line with several clinical studies that suggest not only a lower incidence of TD in schizophrenic patients treated with risperidone, but also an antidyskinetic effect of this drug in patients previously treated with classical neuroleptics. PMID- 12725900 TI - Biochemical markers for post-operative fatigue after major surgery. AB - OBJECTIVE: To establish a link between tryptophan (a precursor for 5 hydroxytryptamine (5-HT) or serotonin, involved in sleep and fatigue) and post operative fatigue after major surgery. BACKGROUND: There is a link between tryptophan (the precursor for the neurotransmitter 5-hydroxytryptamine), and its competitive binding with non-esterified fatty acids (NEFA) to albumin in the blood. An increase in the plasma concentration of free tryptophan can lead to an increased rate of synthesis of 5-HT in the brain. Free tryptophan competes with the branched chain amino acids (BCAA) for the same port of entry across the blood brain barrier. It is suggested that the plasma concentration of these amino acids could be a marker of post-operative fatigue. In a previous study undertaken in this laboratory on patients undergoing two different types of major surgery, similar post-operative increases were observed in the plasma concentration of free tryptophan and the plasma concentration ratio of free tryptophan to branched chain amino acids. However, the study was retrospective and no measure of fatigue had been made. METHODS: In the present study, this deficiency has been addressed by administering a modified Profile of Mood States questionnaire to patients undergoing reconstructive or colorectal surgery. In addition, blood samples were measured for plasma free tryptophan, albumin, NEFA and branched chain amino acids before and on 2 days after surgery. RESULTS: There was a significant correlation between fatigue scores and plasma free tryptophan (P<0.000), and the plasma concentration ratio of free tryptophan/BCAA (P<0.016) after surgery in all the patients studied (n=34). This correlation was more marked in the colorectal surgery patients, in whom surgery was more severe. In the three categories of patients receiving elective reconstructive surgery (n=24), those having breast reductions (n=6) had a lower plasma concentration of NEFA and appeared to recover from fatigue more quickly than those with pre-tibial lacerations or malignant melanoma. CONCLUSIONS: These data provide further evidence of a possible biochemical mechanism for central fatigue which involves a precursor of 5-HT. The provision of branched chain amino acids may help to combat the surge in free tryptophan that occurs during stress such as major surgery. PMID- 12725901 TI - NMDA receptor blockade and hippocampal neuronal loss impair fear conditioning and position habit reversal in C57Bl/6 mice. AB - The interpretation of learning and memory deficits in transgenic mice has largely involved theories of NMDA receptor and/or hippocampal function. However, there is little empirical data that describes what NMDA receptors or the hippocampus do in mice. This research assessed the effects of different doses of the NMDA receptor antagonist, MK-801, or different-sized hippocampal lesions on several behavioral parameters in adult male C57Bl/6 mice. In the first set of experiments, different doses of MK-801 (0.05-0.3mg/kg, s.c.) were assayed in fear conditioning, shock sensitivity, locomotion, anxiety, and position habit reversal tests. Contextual and cued fear conditioning, and position habit reversal were impaired in a dose dependent manner. Locomotor activity was increased immediately after injection of the highest dose of MK-801. A second set of experiments determined the behavioral effects of a moderate and large excitotoxic hippocampal lesion. Both lesions impaired contextual conditioning, while the larger lesion interfered with cued conditioning. Reversal learning was significantly diminished by the large lesion, while the moderate lesion had a detrimental effect at a trend level (P<0.10). These results provide important reference data for studies involving genetic manipulations of NMDA receptor or hippocampal function in mice. Furthermore, they serve as a basis for a non-transgenic mouse model of the NMDA receptor or hippocampal dysfunction hypothesized to occur in human cognitive disorders. PMID- 12725902 TI - Induction of Fos in the accessory olfactory system by male odors persists in female mice with a null mutation of the aromatase (cyp19) gene. AB - The ability of odors from soiled male bedding to induce neuronal Fos immunoreactivity (IR) in sensory neurons located in both the apical and basal zones of the vomeronasal organ (VNO) and in two segments of the VNO-projection pathway, the anterior nucleus of the medial amygdala and the bed nucleus of the stria terminalis (BNST), was significantly reduced in adult, ovariectomized, estrogen-treated female mice with a homozygous null mutation of the cyp19 gene (ArKO) which encodes the estrogen biosynthetic P450 enzyme, aromatase. However, a significant odor-induced activation of Fos-IR was seen in other segments of the VNO-projection pathway of ArKO females, including the accessory olfactory bulb (AOB) granule cell layer, the posterior-dorsal medial amygdala (MePD), and the medial preoptic area (MPA). These results suggest that the VNO/accessory olfactory pathway to the hypothalamus was functional in ArKO females even though they had presumably been exposed to less estrogenic stimulation than wild-type (WT) control females throughout development and until the time that estrogen treatment was begun in adulthood. Thus, the hypothesis of Toran-Allerand [Prog. Brain Res. 61 (1984) 63] that female-typical features of neuroendocrine and behavioral function require perinatal exposure to estrogen was not supported, at least for the VNO/accessory olfactory system. PMID- 12725903 TI - Autonomic and EEG correlates of emotional imagery in subjects with different hypnotic susceptibility. AB - The autonomic and EEG correlates of the response to a cognitive unpleasant stimulation (US) verbally administered to awake hypnotizable and non hypnotizable subjects were studied. They were compared with the values obtained during a resting condition immediately preceding the stimulus and with those produced by a cognitive neutral stimulation (NS), also administered after a basal resting period. Results showed hypnotic trait effects on skin resistance, heart and respiratory rate as well as on EEG theta, alpha, beta and gamma relative power changes. The autonomic and EEG patterns observed indicated different strategies in the task execution for hypnotizable and non hypnotizable subjects and a discrepancy between the autonomic and EEG changes associated to the US in susceptible subjects. Results support dissociation theories of hypnosis and suggest for hypnotizable persons an active mechanism of protection against cardiac hazard. PMID- 12725904 TI - Polyunsaturated fatty acids in the blood of spontaneously or induced muricidal male Wistar rats. AB - Serum levels of several n-6 and n-3 polyunsaturated fatty acids were compared in male Wistar muricidal (Mu) and non-Mu rats. The Mu behavior was either spontaneous or induced by long-term isolation, feeding with a starch-enriched polyunsaturated fatty acid diet (PUFA+S), water restriction, or adrenalectomy (ADX). Arachidonic acid (ARA) levels were lower in diet-induced (PUFA+S) Mu rats than in their non-Mu controls. Total n-6 fatty acid levels were also lower in spontaneously Mu rats than in spontaneously non-Mu rats. Docosahexaenoic acid (DHA) and total n-3 fatty acids levels were lower in rats with isolation-induced Mu behavior. The n-3/n-6 ratio was higher in spontaneously Mu rats than in spontaneously non-Mu rats. The changes in ARA levels were greater than those in DHA levels, possibly due to the higher blood-brain barrier passage of arachidonic acid. The results were analyzed in the light of recent results showing a role of PUFAs in human and animal behavior. PMID- 12725905 TI - The cholinergic stimulation of the central amygdala modifying the tonic immobility response and antinociception in guinea pigs depends on the ventrolateral periaqueductal gray. AB - Tonic immobility (TI), also known as death feigning or animal hypnosis, is a reversible state of motor inhibition that is triggered by postural inversion and/or movement restraining maneuvers but also by repetitive stimulation and pressure on body parts. Our previous studies demonstrated that cholinergic stimulation of the central amygdala (CEA) decreases the duration of TI in guinea pigs. Some reports have demonstrated that electrical or chemical stimulation of the CEA promotes antinociception. Evidence suggests that the CEA performs part of its functions by means of a connection with the ventrolateral periaqueductal gray (vlPAG). In the current study, we investigated the participation of a possible functional and anatomical CEA-vlPAG connection in guinea pigs in the regulation of the TI response and antinociception. Our results showed that the functional CEA-vlPAG connection is essential for the participation of the CEA in the modulation of TI and of antinociception. The reversible exclusion of the vlPAG by means of microinjection of 2% lidocaine blocked the inhibitory effect on TI duration and the antinociceptive effect, as determined by a decrease of the vocalization index (VI) obtained with the administration of carbachol (2.7 nmol/0.2 microl) into the CEA. On the other hand, the exclusion of the CEA by lidocaine did not block the antinociception or the increase in TI induced by microinjection of CCh into the vlPAG. Finally, microinjection of the retrograde neurotracer Fast Blue into the CEA or into the vlPAG demonstrated the existence of a reciprocal anatomical connection between the CEA and vlPAG. PMID- 12725906 TI - Convulsive and postural effects of lesioning the mid-substantia nigra pars reticulata in naive and 6-hydroxydopamine lesioned rats. AB - The subthalamic nucleus is targeted for the treatment of Parkinson's disease. Unilateral lesions improve some aspects of parkinsonism but produce postural abnormalities in animal models but the exact pathways producing these effects remain to be defined. Using a battery of tests we evaluated the effects of lesioning one of the two major subthalamic targets, the substantia nigra pars reticulata in naive and 6-OHDA lesioned rats. Lesions targeting the mid substantia nigra pars reticulata resulted in acute tonic-clonic seizures and intense contralateral rotational asymmetry. During the first month after substantia nigra pars reticulata lesions there was normalisation of the ipsilateral head position bias induced by unilateral 6-OHDA lesions, significant contralateral body axis bias but no significant alteration of apomorphine induced rotation and sensorimotor neglect in 6-OHDA lesioned rats. Combined with our previous data, this suggests that subthalamic projections via the substantia nigra pars reticulata are important in seizures and postural behaviours. Therefore unilateral subthalamotomy probably induces postural deficits in hemiparkinsonian animals via projections involving the substantia nigra pars reticulata. This has implications for patients undergoing subthalamotomy for treatment of Parkinson's disease. PMID- 12725907 TI - Sensorimotor transformations in cortical motor areas. AB - A central problem in motor research has been to understand how sensory signals are transformed to generate a goal-directed movement. This problem has been formulated as a set of coordinate transformations that begins with an extrinsic coordinate frame representing the spatial location of a target and ends with an intrinsic coordinate frame describing muscle activation patterns. Insight into this process of sensorimotor transformation can be gained by examining the coordinate frames of neuronal activity in interconnected regions of the brain. We recorded the activity of neurons in primary motor cortex (M1) and ventral premotor cortex (PMv) in a monkey trained to perform a task which dissociates three major coordinate frames of wrist movement: muscle, wrist joint, and an extrinsic coordinate frame. We found three major types of neurons in M1 and PMv. The first type was termed 'extrinsic-like'. The activity of these neurons appeared to encode the direction of movement in space independent of the patterns of wrist muscle activity or joint movement that produced the movements. The second type was termed 'extrinsic-like with gain modulation'. The activity of these neurons appeared to encode the direction of movement in space, but the magnitude (gain) of neuronal activity depended on the posture of the forearm. The third type was termed 'muscle-like' since their activity co-varied with muscle activity. The great majority of the directionally-tuned neurons in the PMv were classified as 'extrinsic-like' (48/59, 81%). A smaller group was classified as 'extrinsic-like with gain modulation' (7/59, 12%). In M1, the three types of neurons were more equally represented. Our results raise the possibility that cortical processing between M1 and PMv may contribute to a sensorimotor transformation between extrinsic and intrinsic coordinate frames. Recent modeling studies have demonstrated the computational plausibility of such a process. PMID- 12725908 TI - The delta2 glutamate receptor: 10 years later. AB - The orphan glutamate receptor delta2 (GluRdelta2) is predominantly expressed in Purkinje cells and plays a crucial role in cerebellar functions: mice that lack the GluRdelta2 gene display ataxia and impaired synaptic plasticity. However, when expressed alone or with other glutamate receptors, GluRdelta2 does not form functional glutamate-gated ion channels nor does it bind to glutamate analogs. Therefore, the mechanisms by which GluRdelta2 participates in cerebellar functions have been elusive. Studies of mutant mice such as lurcher, hotfoot, and GluRdelta2 knockout mice have provided clues to the structure and function of GluRdelta2. GluRdelta2 has a channel pore similar to that of other glutamate receptors; the channel is functional at least when the lurcher mutation is present. GluRdelta2 must be transported to the Purkinje cell surface to function; the absence of surface GluRdelta2 causes the ataxic phenotype of hotfoot mice. In GluRdelta2-null mice, the presence of naked spines not innervated by parallel fibers may influence the sustained innervation of mutant Purkinje cells by multiple climbing fibers. From these results, several hypotheses about mechanisms by which GluRdelta2 functions are proposed in this article. Further characterization of GluRdelta2's functions will provide key insights into normal and abnormal cerebellar functions. PMID- 12725909 TI - Circadian fluctuation of time perception in healthy human subjects. AB - Previous studies suggested that various psychophysiological factors have influences on human time perception. In particular, working memory loads, time of day, body temperature, and mood were known as important modifiers of time perception. The purpose of this study is to elucidate factors affecting the short term time perception under controlled condition. Fourteen healthy young male adults participated in this study. Time perception sessions (TPS) were conducted 4 times at 0900, 1300, 1700 and 2100 h. The TPS consisted of five 10-s time production trials under five different conditions (control trial, those with reward, and 3 different dual-load working memory tasks). Subjective status was assessed using visual analogue scales (VAS). To verify a participant's vigilance state, an alpha attenuation coefficient (AAC) was calculated. Two-way repeated measures ANOVA for produced time revealed a significant main effect of session, but no effect of task or interaction. Although produced time was not correlated with AACs or VAS scores, there was a significant negative correlation between produced time and core body temperature. These results suggest that human short term time perception may be more influenced by circadian rhythm than working memory load or psychophysiological status. PMID- 12725910 TI - NAC1, a POZ/BTB protein present in the adult mammalian brain, triggers apoptosis after adenovirus-mediated overexpression in PC-12 cells. AB - POZ/BTB proteins influence cellular development and in some examples act as oncoproteins. However, several POZ/BTB transcription factors have been found in terminally differentiated neurons, where their functions remain unknown. One example is NAC1, a constitutively-expressed protein that can regulate behaviors associated with cocaine use. The present study represents an initial attempt to understand the actions of NAC1 within neurons by using adenoviral-mediated gene transfer into differentiated PC-12 cells. Cell survival in PC-12 cells overexpressing NAC1 was greatly reduced compared with cells infected by a control Ad-GFP. The morphological appearance of the dying cells was consistent with programmed cell death. Fragmentation of genomic DNA occurred in PC-12 cells infected with adenoviruses encoding NAC1 but not control viruses. NAC1 over expression was followed by the down regulation of the anti-apoptotic proteins Bcl 2 and Bcl-2-xl. Concurrently, levels of the pro-apoptotic proteins Bax and p53 increased following NAC1 overexpression. These observations suggest that NAC1expression in PC-12 cells induces apoptosis by altering the expression of these upstream mediators of the execution phase of programmed cell death. These findings raise the possibility that aberrantly regulated NAC1 expression in the mammalian brain may contribute to programmed cell death. PMID- 12725911 TI - Overexpression of Ca2+-permeable AMPA receptor promotes delayed cell death of hippocampal CA1 neurons following transient forebrain ischemia. AB - To examine the role of Ca(2+) entry through AMPA receptors in the pathogenesis of the ischemia-induced cell death of hippocampal neurons, we delivered cDNA of Q/R site-unedited form (GluR2Q) of AMPA receptor subunit GluR2 in the hippocampus by using an HVJ-liposome-mediated gene transfer technique. Two days prior to transient forebrain ischemia, we injected an HVJ-liposome containing cDNA of the GluR2Q-myc fusion gene into a rat unilateral hippocampus. In the absence of ischemic insult, overexpression of Ca(2+)-permeable GluR2Q did not cause any neurodegeneration in the cDNA-injected hippocampus. In ischemic rats, overexpression of Ca(2+)-permeable GluR2Q markedly promoted ischemic cell death of CA1 pyramidal neurons, while complete rescue of CA1 pyramidal neurons from ischemic damage occurred in the hippocampal hemisphere opposite the GluR2Q expression. Overexpression of the Q/R-site edited form (GluR2R) of subunit GluR2 did not affect the ischemia-induced damage of CA1 pyramidal neurons. From these results, we suggest that the Ca(2+)-permeability of AMPA receptors does not have a direct contribution to glutamate receptor-mediated neurotoxicity but has a promotive action in the evolution of ischemia-induced neurodegeneration of vulnerable neurons. PMID- 12725912 TI - Morphological study of orexin neurons in the hypothalamus of the Long-Evans rat, with special reference to co-expression of orexin and NADPH-diaphorase or nitric oxide synthase activities. AB - Orexins, novel neuropeptides, are exclusively localized in the hypothalamus and implicated in the regulation of a variety of activities, including food intake and energy balance. Nitric oxide (NO), an unconventional neurotransmitter, is widely present in numerous brain regions including the hypothalamus, and has similar physiological roles to those of the orexins. The present study was undertaken to examine the distribution of orexin neurons and the presence of neuronal nitric oxide synthase (nNOS) in the orexin neurons to clarify whether NO interacts with the orexins in the neuronal regulation activities in the Long Evans rat. We used two double-labeling methods: nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry in combination with orexin immunohistochemistry, and double-labeling fluorescent immunohistochemistry for orexin and nNOS. The majority of the orexin immunoreactive neurons were localized mainly in the areas of the dorsomedial hypothalamic nucleus (DMN), the dorsal part of the perifornical nucleus (PEF) and lateral hypothalamic area. The orexin immunoreactive cell bodies were medium in size, and triangular, round, elliptic, and fusiform in shape. The sizes and shapes of orexin neurons in the different parts were similar. Cell bodies coexpressing the orexin and nNOS or NADPH-d were present in the areas of the DMN and the PEF, and the nerve fibers containing orexin and nNOS were distributed in the DMN and PEF, arcuate nucleus (ARN) and ventromedial hypothalamic nucleus (VMH). These results provide morphological evidence that there exists a population of nNOS- or NADPH-d-/orexin-coexpressing neurons in the orexinergic cell group in the hypothalamus, and taken together with previous findings, suggest that NO may play a role in the mechanisms by which orexin neurons regulate food intake and energy balance. PMID- 12725913 TI - Quantitative age-related changes in NADPH-diaphorase-positive neurons in the ventral lateral geniculate nucleus. AB - Age-related changes in nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) were examined in the rat ventral lateral geniculate nucleus (vLGN) using histochemical methods. Eighteen rats aged 3, 24, and 26 months were studied using quantitative methods to investigate the number of neurons per mm(2), the cross-sectional area, and the orientation of dendritic processes of NADPH-d positive neurons. We have described three types of neurons: types A and B are both located in the lateral and medial vLGN (vLGN-l and vLGN-m, respectively), and type C neurons over the optic tract. The number of NADPH-d-positive neurons was significantly reduced in the old rats (-39%) when compared with controls (3 month-old rats). The quantitative analysis of cell areas revealed a significant decrease of somatic size in type B neurons, both in the lateral and medial vLGN, and in C neurons; however, type A neurons did not show significant changes. By quantifying the orientation of dendritic processes, we observed a predominant dorsolateral orientation in type A and B neurons. During aging, there are no changes in the dendritic orientation of neurons located in the vLGN-m; however, vLGN-l neurons show an increase in dendritic processes with dorsoventral orientation. In type C neurons, our results show that 87.4% of dendritic processes are lateromedially oriented at 26 months old. Therefore, the types A and B neurons behave differently during senescence. Type A neurons do not change in size, but those located in the vLGN-l modify the orientation of their dendritic processes; however, type B neurons, reduce their size and those located in the vLGN-l also modify their dendritic process orientation. Finally, the type C neurons modify their size and dendritic process. PMID- 12725914 TI - Release from GABA(A) receptor-mediated inhibition unmasks interlaminar connection within superior colliculus in anesthetized adult rats. AB - The superior colliculus (SC) contains two major subdivisions, the superficial layers (sSC) and the deeper layers (dSC). sSC receives visual information from the retina and visual cortex, while dSC sends descending projections to the brainstem and spinal cord. It has not been clear whether and how sSC directly activates dSC, however recent studies in slice preparations reported that electrical stimulation of sSC induces burst firing in dSC neurons after application of bicuculline. In the present study, we tested whether sSC directly activates dSC in vivo. In isoflurane-anesthetized rats, electrical stimulation of the optic nerve (ON) induced negative field responses mainly in sSC, but not as much in dSC, under control conditions. However, after injection of bicuculline into dSC, ON stimulation induced long-lasting negative field responses in dSC. dSC neurons including the tectofugal neurons exhibited burst firing during the long-lasting negative field responses. The burst responses remained after ablation of the cerebral cortex ipsilateral to the SC recordings. These results suggest that retinal inputs induce burst responses in tectofugal neurons in dSC via subcortical pathways when the SC circuit is released from GABA(A) receptor mediated inhibition. We propose that sSC is the main candidate for the source of the excitatory synaptic inputs to dSC among the subcortical regions. PMID- 12725915 TI - Great potentiality of neonatal facial motor neurons for neural plasticity as determined by functionally essential neuronal population. AB - The present study was undertaken to determine the neuronal population essential for normal and minimal facial function of young adult rats that had received various degrees of crush injuries to the facial nerve in the neonatal period. Using a neuronal tracer, it was found in young adult rats receiving neonatal injuries that the minimum number of tracer-labeled facial motor neurons necessary for normal facial function corresponded to 13-14% of the neurons (2540+/-64) of the age-matched control animals, whereas the minimum number of neurons necessary for minimal facial function corresponded to 5%. On the other hand, the minimum numbers of tracer-labeled facial motor neurons necessary for normal and minimal facial function of young adult rats that received various degrees of crush injuries corresponded to 61 and 27-30%, respectively, of the neurons (2540+/-64) of the uninjured control animals. These results indicate that the facial function of animals with nerves crushed at the neonatal stage can be adequately maintained by a very small population of neurons, implying a great potential of neonatal neurons for neural plasticity. PMID- 12725916 TI - Resets of torsional eye position errors in different lighting conditions. AB - To investigate a resetting mechanism of torsional eye position errors, spontaneous scanning eye movements and visually guided eye movements in different lighting conditions were recorded three dimensionally. Two monkeys (Macaca fuscata, TS, MI) were engaged in this experiment. A dual scleral search coil method was used for three-dimensional (3-D) eye movement recordings. In complete darkness, the thickness of Listing's plane at the onset of spontaneous saccades (0.51/0.37 degrees (TS/MI)) and that at the end of spontaneous fixation periods (0.47/0.34 degrees (TS/MI)) were significantly (P<0.001) smaller than that at the end of spontaneous saccades (0.59/0.45 degrees (TS/MI)) and that at the onset of spontaneous fixation periods (0.58/0.44 degrees (TS/MI)). Such differences in the thickness of Listing's plane were not observed in the light (P>0.10). Amplitude of torsional drift during post-saccadic fixation period was correlated with the amount of torsional position error at the end of saccade (P<0.001). The slope of regressed line in the dark or dim light (-0.40 to -0.50/-0.32 to -0.33 (TS/MI)) was steeper than that in the light (-0.04/-0.03 to -0.09 (TS/MI)). A resetting mechanism for torsional eye position errors during post-saccadic fixation periods is active in the dark but inactive in the light. PMID- 12725917 TI - Cloning and functional characterization of squid voltage-dependent Ca2+ channel beta subunits: involvement of N-terminal sequences in differential modulation of the current. AB - cDNAs that encode beta subunits of voltage-dependent Ca(2+) channel were cloned from the optic lobe of the squid Loligo bleekeri. The subunits, LoCa(v)beta(1a) and LoCa(v)beta(1b) are 96% identical in amino acid sequence. The sole sequence differences are in the N-terminal region and in a five amino acid insertion in the central region of LoCa(v)beta(1b). RT-PCR revealed that LoCa(v)beta(1a) and LoCa(v)beta(1b) transcripts were expressed mainly in the optic lobe and stellate ganglion, and more weakly in mantle muscle, systemic heart, gill, branchial heart, stomach and liver. Coexpression of LoCa(v)beta(1a) or LoCa(v)beta(1b) with mammalian Ca(v)2.3 and alpha(2)/delta subunits in the Xenopus oocyte resulted in high-voltage-activated currents, and showed slow current inactivation and moderate steady-state inactivation. Comparison of the squid subunits with four mammalian beta subunits, beta(1b), beta(2a), beta(3) and beta(4), demonstrated that the modulatory effects of the beta subunits on steady-state inactivation kinetics were beta(3)0.05 ANOVA). Both materials exhibited fracture toughness anisotropy following pressing. They demonstrated improved fracture toughness and reduced hardness compared with IPS-Empress P<0.05(ANOVA), which should be beneficial for clinical applications. PMID- 12726703 TI - Effect of light power density variations on bulk curing properties of dental composites. AB - OBJECTIVE: The hypothesis that low light intensity and long but sufficient curing time can produce composites with volumetric shrinkage, degree of conversion (DC%) and Young's modulus (E-modulus) comparable to those of high light intensity cured composite was tested, when the contraction strain and heat generation were lower with low light intensity curing. METHODS: Dental composites (Z100 and Z250, 3M ESPE) were investigated. Specimens were cured with light intensities of 200, 450 and 800 mW/cm(2) for 140, 60 and 35 s from a distance of 7 mm. Strain-gages were used for contraction strain measurements. DC% was measured at the top and the bottom of 4 mm thick samples using FT-Raman spectroscopy. Volumetric polymerization shrinkage was determined using a water displacement method. E modulus was determined in tension on composite specimens. RESULTS: The results were analyzed using ANOVA and Duncan's multiple range tests and regular t-test. Polymerization stress level decreased significantly (p<0.05) when cured with 200 mW/cm(2) rather than with 800 mW/cm(2). Temperature rises were significantly different (p<0.05) for different composites and light intensity values. Reduction in light intensity did not decrease the DC% values significantly at the top surfaces. The most dramatic differences existed between top and bottom surfaces (p<0.05) rather than among curing groups. Measured E-modulus and volumetric shrinkage values were not significantly different (p>0.05) between different light intensity groups. CONCLUSION: DC%, E-modulus and the volumetric shrinkage values in cured composites were not affected by low light intensity, however, the contraction strain and polymerization's exotherm were decreased. Thus our results support the proposed hypothesis. PMID- 12726704 TI - Radiographic equipment and techniques used in general dental practice: a survey of general dental practitioners in England and Wales. AB - OBJECTIVES: The aims of this study were to determine the self-reported use of panoramic radiography, D- and E-speed film, rectangular collimation, film holders, equipment fitted with a long spacer cone (>200 mm) and the bisecting angle and paralleling techniques by general dental practitioners and to see if use was related to the dentists' age and postgraduate qualifications. METHODS: Three mailings of a self-completion questionnaire were circulated to 800 general dental practitioners working in the National Health General Dental Service in England and Wales. RESULTS: A response rate of 74% was achieved. Sixty-one percent of general dental practitioners reported use of panoramic equipment. Fifty percent of dentists always used E-speed film and 18% always used rectangular collimation. Sixty-eight percent of dentists always used bitewing film holders though fewer (37%) used periapical film holders. Sixty-three percent of dentists always used a long cone. The bisecting angle technique was used by 70% of dentists with 22% always using this technique for periapical radiographs. Thirty-one percent always used the paralleling technique. Use of panoramic equipment, periapical film holders, bisecting angle and paralleling techniques were associated with the dentists' age. Use of periapical film holders, bisecting angle and paralleling techniques and rectangular collimation were associated with dentists' postgraduate qualifications. CONCLUSIONS: There are several features of radiographic equipment and techniques that can aid high quality imaging and reduce patient dose. Although a number of these are being used in general dental practice in England and Wales they have yet to achieve universal adoption. PMID- 12726705 TI - Shear bond strength of aesthetic materials bonded to Ni-Cr alloy. AB - OBJECTIVES: This study was undertaken to evaluate the shear bond strength of four materials used as aesthetic material bonded to Ni-Cr alloy. METHODS: Sixty-eight alloy discs were prepared and divided equally into four groups, and received four treatments for veneering: conventional feldspathic porcelain (Noritake EX-3) and three light-cured prosthodontic composite resins (Artglass, Solidex and Targis). The aesthetic materials were applied after metal structure conditioning in accordance with the manufacturers' recommendations. The specimens were stored in distilled water at 37 degrees C for 7 days. A universal testing machine was used to measure the shear bond strength of the specimens at a cross head speed of 0.5 mm/min. Fractured specimens were examined by using both optical and scanning electron microscope. RESULTS: The analysis of variance and Tukey's test showed that the strongest mean shear bond was obtained with Noritake EX-3 (mean shear bond strength 42.90+/-7.82 MPa). For composites, the highest mean shear bond strength was observed for Targis (12.30+/-1.57 MPa); followed by Solidex (11.94+/ 1.04 MPa) and Artglass (10.04+/-0.75 MPa). Optical analysis of the fractured surfaces indicated that for Targis and Noritake EX-3 all failures were a mixture of both cohesive and adhesive patterns. As for Artglass and Solidex, the fractures were mainly adhesive in nature. CONCLUSIONS: The Solidex system was equivalent to the Targis system in bond strength and exhibited greater strength than the Artglass system. The porcelain fused-to-metal showed considerably higher shear bond strength than the three metal-resin bonding techniques. PMID- 12726706 TI - Surface modification of an experimental silicone rubber maxillofacial material to improve wettability. AB - OBJECTIVES: Good wettability of maxillofacial prosthetic materials is important so that a lubricating layer is formed with supporting tissues thus reducing patient discomfort. The purpose of the study was to surface modify an experimental silicone rubber material in order to improve wettability. METHODS: Samples of experimental silicone rubber were surface modified by first argon plasma treatment followed by chemisorption of ethyleneoxy functional silanes. These were compared with the same silicone rubber which had ethyleneoxy functional surfactants incorporated into the polymer matrix. In all cases contact angles, tear strength and water uptake were measured. RESULTS: Surface modified materials had comparable contact angles to surfactant modified silicone rubber all being significantly lower than the unmodified material. Surface modified materials, however, had a significantly higher tear strength and lower water uptake in comparison to surfactant modified materials. CONCLUSIONS: Argon plasma treatment followed by chemisorption of ethyleneoxy functional silanes proved an effective way of improving the wettability of an experimental silicone rubber maxillofacial prosthetic material without altering bulk properties. PMID- 12726707 TI - Dental services and perceived oral health: are patients better off going private? AB - OBJECTIVE: The aim of this study is to determine associations between method of payment for dental services and perceived oral health in the UK. METHOD: A national UK study involving a random probability sample of 2718 adults. Respondents were interviewed in their homes about their method of payment for dental services (private or NHS), service use (time and reason for last dental visit), self-reported oral health status (number of teeth possessed and denture status) and the impact of their oral health on their quality of life (employing the 16-item OHQoL-UK( Copyright) measure). RESULTS: The response rate was 68% (1838/2718). Thirty one percent (575/1838) claimed they paid privately for dental services the last occasion they visited their dentist. This was associated with self-reported number of teeth possessed (P<0.01), denture status (P<0.01), WHO goal of retaining 20 teeth with and without a prosthesis (P<0.05), impact of oral health on life quality (P<0.01), and number of positive oral health influences experienced (P<0.01). However, these associations did not remain apparent having accounted for socio-demographic factors (age, gender, and social class background) and reported dental attendance pattern. CONCLUSION: Difference in perceived oral health exists between private and NHS dental service users. However, this is more likely to be attributed to socio-demographic factors and regular use of services rather than method of payment. PMID- 12726708 TI - Cytotoxicity of three dentin bonding agents on human dental pulp cells. AB - OBJECTIVES: Dentin bonding agents (DBA) have been widely used in operative restoration to prevent leakage and promote bonding strength in the resin-dentin interface. However, DBA may exert potentially harmful effects to the dental pulp. In the present study, differential cytotoxicity of three DBA (Syntac Sprint, SP; Prime and Bond 2.1, PB; and Single Bond, SB) on the pulp cells was tested. METHODS: Three DBA were diluted with the culture medium by a ratio of 1:1000, 1:2000 and 1:4000 (v/v). Pulp cells (5 x 10(4) cells/well) were then exposed to culture medium containing different diluents of three DBA for 12, 24h and 3 days. Cytotoxicity was measured with a modified 3-(4,5-dimethyl-thiazol-2-yl)-2,5 diphenyl-tetrazolium bromide (MTT) assay. RESULTS: A 12h experiment revealed that SP was the strongest cytotoxic agent, followed sequentially by SB and PB. Exposure of pulp cells to 1:4000 (v/v) dilution of SP, PB and SB for 24h reduced the cell number by 23, 6 and 45%, respectively. A 1:2000 (v/v) of DBA diluents reduced the cell number for 32, 13 and 65%, respectively, by SP, PB and SB. Dilution of DBA by 1000-fold of culture medium further enhanced the cytotoxic response. Cell number decreased by 89, 65 and 72%, respectively, by SP, PB and SB. Similar to the 12h-cytotoxicity data, SB is more toxic at high dilution condition, whereas, at low dilution condition, SP is the most toxic agent to pulp cells. Similar cytotoxicity was noted when pulp cells were exposed to DBA for 3 days. Toxicity of DBA was concomitant with marked retraction and rounding of dental pulp cells. SIGNIFICANCE: These results indicate that DBA exerts potential harmful effects to the pulp. Differential toxic effects of DBA on the pulp cells should be considered during selection of a suitable DBA for operative restoration. PMID- 12726710 TI - Macrophage response to cross-linked and conventional UHMWPE. AB - To prevent wear debris-induced osteolysis and aseptic loosening, cross-linked ultra-high molecular weight polyethylene's (UHMWPE) with improved wear resistance have been developed. Hip simulator studies have demonstrated very low wear rates with these new materials leading to their widespread clinical use. However, the biocompatibility of this material is not known. We studied the macrophage response to cross-linked UHMWPE (XLPE) and compared it to conventional UHMWPE (CPE) as well as other clinically used orthopaedic materials such as titanium alloy (TiAlV) and cobalt-chrome alloy (CoCr). Human peripheral blood monocytes and murine macrophages, as surrogates for cells mediating peri-implant inflammation, were cultured onto custom designed lipped disks fabricated from the test materials to isolate cells. Culture supernatants were collected at 24 and 48h and analyzed for cytokines such as IL-1alpha, IL-1beta, TNF-alpha and IL-6. Total RNA was extracted from adherent cells and gene expression was analyzed using qualitative RT-PCR. In both in vitro models, macrophages cultured on cross linked and conventional polyethylene released similar levels of cytokines, which were also similar to levels on control tissue culture dishes. Macrophages cultured on TiAlV and CoCr-alloy released significantly higher levels of cytokines. Human monocytes from all donors varied in the magnitude of cytokines released when cultured on identical surfaces. The variability in individual donor responses to TiAlV and CoCr surfaces may reflect how individuals respond differently to similar stimuli and perhaps reveal a predisposed sensitivity to particular materials. PMID- 12726711 TI - Sodium silicate gel as a precursor for the in vitro nucleation and growth of a bone-like apatite coating in compact and porous polymeric structures. AB - In the present work, a new methodology to produce bioactive coatings on the surface of starch-based biodegradable polymers or other polymeric biomaterials is proposed. A sodium silicate gel is employed as an alternative nucleating agent to the more typical bioactive glasses for inducing the formation of a calcium phosphate (Ca-P) layer. The method has the advantage of being able to coat efficiently both compact materials and porous 3D architectures aimed at being used on tissue replacement applications and as tissue engineering scaffolds. By means of this treatment, it is possible to observe the formation of an apatite like layer, only after 6 hours of simulated body fluid immersion. For the porous materials, this layer could also be observed inside the pores, clearly covering the cell walls. Furthermore, an increase of the surface hydrophilicity (higher amount of polar groups in the surface) might contribute to the formation of silanol groups that also act as apatite inductors. After 30 days of SBF immersion, the apatite-like films exhibit a partially amorphous nature and the Ca/P ratios became much closer to the value attributed to hydroxyapatite (1.67). The obtained results are very promising for the development of cancellous bone replacement materials and for pre-calcifying bone tissue engineering scaffolds. PMID- 12726712 TI - Characterization of chemical inhomogeneity in plasma-sprayed hydroxyapatite coatings. AB - Successful applications of plasma-sprayed hydroxyapatite (HA) coating for implants rely on understanding characteristics of the coating's microstructure, particularly its inhomogeneity. We explored three new techniques for characterizing the chemical inhomogeneity of sprayed HA coatings on titanium substrate: micro-Raman spectroscopy (MRS), positive and negative ion ratios of time-of-flight secondary ion mass spectrometry (ToF-SIMS) and the energy loss peaks of X-ray photoelectron spectroscopy (XPS). The results showed that MRS effectively revealed a chemical gradient in the direction of the coating thickness and a decrease in crystallinity from the surface to interface within the as-sprayed coatings. The post-spray treatment effectively promoted homogeneity between surface and the coating/Ti interfaces. Elucidating the chemistry of the sprayed HA coatings using the ion ratios of ToF-SIMS and the energy loss peaks of XPS remains a challenge, even though such techniques can be used to identify certain calcium phosphate phases in pure powder form. PMID- 12726713 TI - Comparison of human bone marrow stromal cells seeded on calcium-deficient hydroxyapatite, beta-tricalcium phosphate and demineralized bone matrix. AB - The aim of this study was to compare three resorbable biomaterials regarding seeding efficacy with human bone marrow stromal cells (BMSCs), cell penetration into the matrix, cell proliferation and osteogenic differentiation. Calcium deficient hydroxyapatite (CDHA), beta-tricalcium phosphate (beta-TCP), and demineralized bone matrix (DBM) were seeded with human BMSCs and kept in human serum and osteogenic supplements for 3 weeks. Morphologic and biochemical evaluations were performed on day 1, 7, 14 and 21. The allograft DBM and CDHA exhibited both an excellent seeding efficacy while the performance of beta-TCP was lower when compared. The total protein content and the values for specific alkaline phosphatase (ALP) increased on all matrices and no significant difference was found for these two markers. BMSCs in monolayer had a significant increase of protein, but not of ALP. Osteocalcin (OC) values increased significantly higher for BMSC in cultures on DBM when compared to CDHA and beta TCP. The OC levels decreased significantly in the BMSC monolayer culture. BMSCs were found inconsistently within the synthetic materials, whereas in DBM they were found more homogeneously distributed throughout the matrix. All three matrices promoted BMSC proliferation and differentiation to osteogenic cells. DBM allografts seem to be more favorable with respect to cell ingrowth tested by histology, and osteogenic differentiation ascertained by an increase of OC. CDHA with its high specific surface area showed more favorable properties than beta TCP regarding reproducibility of the seeding efficacy. PMID- 12726715 TI - Radio frequency (RF) suspension plasma sprayed ultra-fine hydroxyapatite (HA)/zirconia composite powders. AB - Ultra-fine hydroxyapatite (HA)/ZrO(2) composite powders was synthesised by radio frequency (RF) induction suspension plasma spray. A wet suspension of HA/ZrO(2) was employed as feedstock. The suspension was injected axially into the RF plasma to produce the nano-composite powders, which were subsequently accumulated in cyclone collectors. The particle size and morphology was resolved by using the Zeta potential nano-particle size analyser, scanning electron microscopy, transmission electron microscopy, field emission microscopy techniques. The phase composition, phase concentration, and, molecular structure of the powders were characterised using differential scanning calorimetry, Fourier transform infra red and X-ray diffractometry with quantitative phase analysis empowered by the Rietveld method. Results indicated that nano-size, spherical HA/ZrO(2) composite powders were produced with varying morphological features that depend on the thermal treatment. Calcium zirconate (CaZrO(3)) was produced as a byproduct whose biocompatibility is not well documented. Results also showed that the HA decomposed into alpha and beta-TCP due to decreasing Ca/P ratio with the formation of CaZrO(3). PMID- 12726714 TI - Hepatocyte performance on different crystallographic faces of rutile. AB - The influence of crystallographic orientation of polished rutile single crystal surfaces of the (100), (110) and (111) orientation on hepatocyte performance was tested in cell culture over 3 days. Cell adhesion was observed on the titanium dioxide surfaces and their performance was measured by means of cell number attached (protein mass), cell viability (neutral red assays) and metabolic activity (thiazolyl tetrazolium bromide assay). Titanium dioxide displays no cytotoxic effects on hepatocytes, and shows a performance in the range of standard collagen-coated tissue culture polystyrene dish. The number of hepatocytes adhered on the different rutile surfaces were not significantly different to those on dense rutile polycrystalline ceramic. These findings suggest that hepatocytes do not recognize the specific differences of differently orientated rutile crystal surfaces. PMID- 12726716 TI - In vivo dissolution behavior of various RF magnetron-sputtered Ca-P coatings on roughened titanium implants. AB - RF magnetron sputter deposition was used to produce 0.1, 1.0 and 4.0 microm thick Ca-P coatings on TiO(2)-blasted titanium discs. Half of the as-sputtered coated specimens were subjected to an additional infrared heat treatment for 30s at 425 475 degrees C. X-ray diffraction demonstrated that infrared radiation changed the amorphous 4 microm sputtered coatings into an amorphous-crystalline structure, while the amorphous 0.1 and 1 microm changed in a crystalline apatite structure with the presents of tetracalciumphosphate as a second phase. Scanning electron microscopically examination of the sputtered coatings revealed that annealing of the 4 microm thick coatings resulted in the appearance of small cracks. Subsequently, the discs were implanted subcutaneous into the back of rabbits. After 1, 4, 8 and 12 weeks of implantation, the implants were retrieved and prepared for histological and physicochemical evaluation. Histological evaluation revealed that the tissue response to all coated implants was very uniform. A very thin connective tissue capsule surrounded all implants. The capsule was usually free of inflammatory cells. At the interface, there was a close contact between the capsule and implant surface and no inflammatory cells were seen. Physicochemical evaluation showed that the 0.1 and 1 microm thick amorphous coatings had disappeared within 1 week of implantation. On the other hand, the 4 microm thick amorphous phase disappeared during the implantation periods, which was followed by the precipitation of a crystalline carbonate apatite. Further, at all implantation periods the heat-treated 1 and 4 microm thick coatings could be detected. Occasionally, a granular precipitate was deposited on the heat-treated 4 microm thick coating. Fourier transform infrared spectroscopy showed the formation of carbonate apatite (CO(3)-AP) on the 4 microm thick amorphous coating and on the heat-treated specimens. On basis of our findings, we conclude that 1 microm thick heat-treated Ca-P sputter coating on roughened titanium implants appear to be of sufficient thickness to show bioactive properties, under in vivo conditions. PMID- 12726717 TI - Transforming growth factor beta1 immobilized adsorptively on Ti6Al4V and collagen type I coated Ti6Al4V maintains its biological activity. AB - Titanium and titanium alloys are often used for orthopedic and dental implants. Osseointegration of Ti6Al4V may be improved not only by precoating of the surface with extracellular matrix proteins like collagen type I but also by additional immobilization of growth factors. In the present study, transforming growth factor beta1 (TGF-beta1) which is known as an inducer of collagen synthesis was immobilized adsorptively on uncoated and collagen type I coated Ti6Al4V surfaces. TGF-beta1 was found immobilized slightly faster to collagen type I coated than to uncoated Ti6Al4V and released slower from the collagen coated material. Immobilized TGF-beta1 is biologically active for at least 3 weeks storage at 4 degrees C. Sterilization by ethylene oxide inactivates immobilized TGF-beta1. In osteoblasts cultured on implants with adsorptively immobilized TGF-beta1, mRNA level and specific catalytic activity of alkaline phosphatase as well as accumulation of calcium and phosphate were found reduced, whereas procollagen alpha1(I) mRNA level and the rate of collagen synthesis were increased. PMID- 12726718 TI - Poly(ether ester amide)s for tissue engineering. AB - Poly(ether ester amide) (PEEA) copolymers based on poly(ethylene glycol) (PEG), 1,4-butanediol and dimethyl-7,12-diaza-6,13-dione-1,18-octadecanedioate were evaluated as scaffold materials for tissue engineering. A PEEA copolymer based on PEG with a molecular weight of 300 g/mol and 25wt% of soft segments (300 PEEA 25/75) and the parent PEA polymer (0/100) sustain the adhesion and growth of endothelial cells. The in vivo degradation of melt-pressed PEEA and PEA discs subcutaneously implanted in the back of male Wistar rats was followed up to 14 weeks. Depending on the copolymer composition, a decrease in intrinsic viscosity of about 20-30% and mass loss up to 12% were measured. During the degradation process, erosion of the surface was observed by scanning electron microscopy and light microscopy. The thermal properties of the polymers during degradation were measured by differential scanning calorimetry. During the first 2 weeks, a broadening of the melting endotherm was observed, as well as an increase in the heat of fusion. Porous matrices of PEEAs and PEA could be prepared by molding mixtures of polymer and salt particles followed by leaching of the salt. PMID- 12726720 TI - Performance study of braided carbon/PEEK composite compression bone plates. AB - In addition to unidirectional laminates and short fiber reinforcements for compression bone plate developments in the literature, we have proposed using a textile structure, i.e. braid preform, for this purpose. In the present paper, the influence of braiding angles and plate thicknesses on the bending performance of the braided composite bone plates is investigated. As a result, the influence of the braiding angle, varied in a certain range, on the plate bending properties is not significant when the plate thickness is thin. This influence becomes higher with an increase in the plate thickness. A 10 degrees braiding angle has been seen to be appropriate for all the cases under consideration. The present study indicates that the braided composite plate with 2.6mm thickness can be suitable for forearm treatment whereas the braided composite plate of 3.2mm thickness is applicable to femur or tibia fixation. PMID- 12726719 TI - The use of absorbable co-polymer pads with alginate and cells for articular cartilage repair in rabbits. AB - PURPOSE: To determine the effect of polyglycolic acid (PGA)-polylactic acid (PLA) co-polymer pads with calcium alginate on chondrogenic gene expression for chondrocytes cultured in vitro. We also evaluated the ability of these absorbable pads with alginate to deliver chondrocytes and influence osteochondral defect repair in vivo in immature rabbit knees. METHODS: Rabbit rib chondrocytes were suspended in calcium alginate and co-polymer pads composed of either 47.5/52.5 PGA-PLA or 90/10 PGA-PLA at two different cell concentrations and cultured in vitro for 1, 3, and 5 days. Analysis was performed using RT-PCR for chondrogenic gene expression of aggrecan, type II collagen, and type I collagen. Cells labeled with a traceable green fluorescent protein (GFP) marker in vitro were suspended within the pads to analyze for dispersion and attachment to the pad. An in vivo study was performed in which full-thickness (3x4mm(2)) osteochondral defects were made in 60 rabbit knees. The study comprised four treatment groups based on the type of implant into the defect (empty, alginate alone, or either type of co polymer pad) and harvested at either 6 or 12 weeks. Two independent blinded observers analyzed and scored the defects grossly and histologically. RESULTS: In vitro analysis of the chondrocytes after 1, 3, and 5 days in culture showed no statistical differences between the types of PGA/PLA co-polymer pad with regard to expression of aggrecan, type II collagen, or type I collagen. However, although statistically insignificant, the expression of aggrecan and type II collagen was found to be greater than that for type I collagen in both types of pads, confirming the chondrogenic effect of suspension culture for this system. The addition of alginate to polymer pads allowed costal chondrocytes to be implanted in vivo, as evidenced by the attachment of the cells to the fibers and the uniform dispersion of the GFP-labeled cells through the pad as seen on fluorescent microscopy. Histologic results showed improved scores for the 47.5/52.5 PGA-PLA group (21.3) and the 90/10 PGA-PLA group (18.3) when compared to empty (15.3) or alginate alone (15.1) defects at 12 weeks. CONCLUSION: The addition of calcium alginate to the co-polymer pads offers a new approach to deliver cells to an osteochondral defect and may enhance cartilage regeneration. Future application of this model may allow for an arthroscopic delivery system to assist the healing of cartilage defects. PMID- 12726721 TI - Intragenic HIV-1 env sequences that enhance gag expression. AB - Expression of HIV-1 genes is regulated at multiple levels including the complex RNA splicing and transport mechanisms. Multiple cis-acting elements involved in these regulations have been previously identified in various regions of HIV-1 genome. Here we show that another cis-acting element was present in HIV-1 env region. This element enhanced the expression of Gag when inserted together with Rev response element (RRE) into a truncated HIV-1 genome in the presence of Rev. The enhancing activity was mapped to a 263-bp fragment in the gp41 region downstream to RRE. RNA analysis showed that it might function by promoting RNA stability and Rev-dependent RNA export. The enhancement was specific to Rev dependent expression, since it did not enhance Gag expression driven by Sam68, a cellular protein that has been shown to be able to substitute for Rev in RNA export function. PMID- 12726722 TI - Identification of the host determinant of two prolate-headed phages infecting Lactococcus lactis. AB - A gene responsible for host determination was identified in two prolate-headed bacteriophages of the c2 species infecting strains of Lactococcus lactis. The identification of the host determinant gene was based on low DNA sequence homology in a specific open reading frame (ORF) between prolate-headed phages with different host ranges. When a host carrying this ORF from one phage on a plasmid was infected with another phage, we obtained phages with an altered host range at a frequency of 10(-6) to 10(-7). Sequencing of phage DNA originating from 10 independent single plaques confirmed that a genetic recombination had taken place at different positions between the ORF on the plasmid and the infecting phage. The adsorption of the recombinant phages to their bacterial hosts had also changed to match the phage origin of the ORF. Consequently, it is concluded that this ORF codes for the host range determinant. PMID- 12726723 TI - Gradual development of the interferon-gamma response of swine to porcine reproductive and respiratory syndrome virus infection or vaccination. AB - Infection of swine with virulent porcine reproductive and respiratory syndrome (PRRS) virus induced a rapid, robust antibody response that comprised predominantly nonneutralizing antibodies and waned after approximately 3 months. In contrast, the initial onset of virus-specific interferon (IFN)-gamma-secreting cells (SC) in the pig lymphocyte population remained at a fairly low level during this period and then increased gradually in frequency, plateauing at 6 months postinfection. A similar polarization of the host humoral and cellular immune responses was also observed in pigs immunized with a PRRS-modified live virus (MLV) vaccine. Even coadministration of an adjuvant that enhanced the immune response to a pseudorabies (PR) MLV vaccine failed to alter the induction of PRRS virus-specific IFN-gamma SC (comprising predominantly CD4/CD8 alpha double positive memory T cells with a minority being typical CD4(-)/CD8 alpha beta(+) T cells) and the generation of neutralizing antibodies. Moreover, unlike inactivated PR virus, nonviable PRRS virus did not elicit virus-neutralizing antibody production. Presumably, an intrinsic property of this pathogen delays the development of the host IFN-gamma response and preferentially stimulates the synthesis of antibodies incapable of neutralization. PMID- 12726724 TI - Insertion of a foreign sequence on capsid surface loops of human papillomavirus type 16 virus-like particles reduces their capacity to induce neutralizing antibodies and delineates a conformational neutralizing epitope. AB - The aims of this study were to generate chimeric human papillomavirus (HPV)-16 L1 virus-like particles (VLPs) in order to identify immunogenic domains and conformational neutralizing epitopes, and to characterize the regions where a foreign epitope could be introduced. We hypothesized that these regions could be on L1 protein loops since they are exposed on the surface of VLPs. The aims of this study were achieved by mutating HPV-16 L1 proteins. Six amino acids encoding for the epitope 78-83 (DPASRE) of the hepatitis B core (HBc) antigen were introduced within the different loops of the L1 protein at positions 56/57, 140/141, 179/180, 266/267, 283/284 or 352/353. All these chimeric L1 proteins were capable of self-assembly into VLPs. The antigenicity and immunogenicity of some of these VLPs were reduced compared to the levels observed with wild-type VLPs. All were nevertheless able to induce neutralizing antibodies. VLPs with insertion at position 266/267 induced lower levels of neutralizing antibodies, suggesting the involvement of residues situated on FG loop in L1 neutralizing epitopes. All the chimeric L1 proteins except the one with insertion at position 56/57 were also able to induce anti-HBc antibodies, thus suggesting exposure of the HBc epitope on the VLP surface. Taken together, our findings indicate the possibility of designing HPV-derived vectors that are less immunogenic and suggest positions for insertion of defined immune epitopes or cell ligands into L1 protein to be exposed on the surface of VLPs. PMID- 12726725 TI - Specific G2 arrest of caprine cells infected with a caprine arthritis encephalitis virus expressing vpr and vpx genes from simian immunodeficiency virus. AB - Primate lentivirus (HIV and SIV) vpr accessory genes encode 12- to 14-kDa proteins which induce cell cycle arrest at the G2 phase of infected cells, preventing them from going through mitosis. Members of the HIV-2/SIVmac/SIVsmm group also encode a second closely related accessory protein called Vpx. Vpx and HIV Vpr are critical for virus replication in nondividing cells due to their participation in nuclear import of the preintegration complex. Caprine arthritis encephalitis virus (CAEV) and maedi visna virus are the natural lentiviruses of domestic goat and sheep, respectively, and their genomes do not carry vpr and vpx genes. In this study, we generated chimeric CAEV-based genomes carrying vpr and vpx genes from SIVmac239 and tested their ability to induce G2 cell cycle arrest in infected caprine cells. CAEV-pBSCAvpxvpr is the chimeric genome that was shown to be infectious and replication competent. Our data demonstrated that CAEV pBSCAvpxvpr-infected goat synovial membrane cell monolayer developed more cytopathic effects and a high proportion of cells remained in the G2 phase of cell cycle. This G2 arrest was observed both at the early and at the late stages of infection, while minimal effect was observed with the parental CAEV-pBSCA. These results, described for the first time in mammalian cells other than those of primates, indicate that Vpr-induced G2 cell cycle arrest is not restricted to only primate cells. Thus, conservation of Vpx/Vpr protein functions in caprine cells suggests a possible role for these proteins in the virus life cycle and its ability to adapt to new hosts. The data presented here thus raise a pertinent question about the biological significance of the conservation of Vpr and Vpx functions in caprine cells despite the high phylogenic distance between primates and small ruminants. PMID- 12726726 TI - Infection with the oncogenic human papillomavirus type 59 alters protein components of the cornified cell envelope. AB - Infection of the genital tract with human papillomaviruses (HPVs) leads to proliferative and dysplastic epithelial lesions. The mechanisms used by the virus to escape the infected keratinocyte are not well understood. Infection of keratinocytes with HPV does not cause lysis, the mechanism used by many viruses to release newly formed virions. For HPV 11, a type associated with a low risk of neoplastic disease, the cornified cell envelope (CCE) of infected keratinocytes is thin and fragile, and transcription of loricrin, the major CCE protein, is reduced. The effects of high-risk HPV infection on components of the CCE have not been previously reported. HPV 59, an oncogenic genital type related to HPV types 18 and 45 was identified in a condylomata acuminata lesion. An extract of this lesion was used to infect human foreskin fragments, which were grown in athymic mice as xenografts. Continued propagation using extracts of xenografts permitted growth of additional HPV 59-infected xenografts. CCEs purified from HPV 59 infected xenografts displayed subtle morphologic abnormalities compared to those derived from uninfected xenografts. HPV 59-infected xenografts revealed dysplastic-appearing cells with mitotic figures. Detection of loricrin, involucrin, and cytokeratin 10 was reduced in HPV 59-infected epithelium, while small proline-rich protein 3 (SPR3) was increased. Reduction in loricrin was most apparent in regions of epithelium containing abundant HPV 59 DNA. Compared to uninfected epithelium, loricrin transcription was decreased in HPV 59-infected epithelium. We conclude that HPV 59 shares with HPV 11 the ability to alter CCE components and to specifically reduce transcription of the loricrin gene. Because loricrin is the major CCE protein, a reduction in this component could alter the physical properties of the CCE, thus facilitating virion release. PMID- 12726727 TI - Human immunodeficiency virus type 1 envelope-mediated neuropathogenesis: targeted gene delivery by a Sindbis virus expression vector. AB - Sindbis virus (SIN) expression vectors offer the opportunity for studying neuropathogenesis because of their distinct neural cell tropism. Here, we demonstrate that a recombinant SIN vector expressing EGFP (SINrep5-EGFP) infected multiple cell types including neural cells from several species relevant to lentivirus pathogenesis with high levels of transgene expression. Infection of human neurons by a recombinant SIN (SINrep5-JRFL) expressing the full-length envelope from a neurovirulent human immunodeficiency virus type 1 (HIV-1) strain (JRFL) caused increased cytotoxicity compared to infection with SINrep5-EGFP (P < 0.001), while no cytotoxicity was observed among infected human astrocytes or monocytoid cells. Both human monocyte-derived macrophages (MDM) (P < 0.01) and astrocytes (P < 0.001) infected with SINrep5-JRFL released soluble neurotoxins in contrast to SINrep5-EGFP or mock-infected cells, although this was most prominent for the astrocytes. Implantation of SINrep5-JRFL into the brains of SCID/NOD mice induced neuroinflammation, neuronal loss, and neurobehavioral changes characteristic of HIV-1 infection, which were not present in SINrep5-EGFP or mock infected animals. Thus SIN expression vectors represent novel tools for studying in vitro and in vivo HIV-1 neuropathogenesis because of their high levels of transgene expression in specific cell types within the brain. PMID- 12726728 TI - Nitric oxide and HSV vaginal infection in BALB/c mice. AB - Here we study the role of nitric oxide in the vaginal infection of Balb/c mice with herpes simplex virus type 2. Inducible nitric oxide synthase (iNOS) mRNA was detected by RT-PCR in vaginal tissue and inguinal lymph nodes early postinfection. iNOS was also found to be activated in cells recovered from vaginal washings of infected animals. Animals treated with aminoguanidine (AG), an iNOS inhibitor, showed a dose-dependent increase in vaginal pathology after viral infection compared to controls. Viral titers in vaginal washings and vaginas were higher in AG-treated mice. Treated animals presented higher PMN counts in vaginal washings compared to controls. Histopathology studies revealed a profound inflammatory exudate in vaginal tissue of treated animals. Finally, RT PCR analysis showed increased expression of the chemokines MIP-2 and RANTES in vaginal tissue and inguinal lymph nodes of these animals. PMID- 12726729 TI - Naturally occurring amino acid substitutions in the HIV-2 ROD envelope glycoprotein regulate its ability to augment viral particle release. AB - The envelope glycoprotein of HIV-2 ROD10 has the intriguing ability to enhance the rate of viral particle release from infected cells. However, not all HIV-2 envelope glycoproteins are active in this regard. Indeed, we have previously noted that, despite a high degree of identity with that of ROD10, the envelope protein of the ROD14 isolate was unable to enhance virus production. In this study, site-directed mutagenesis was employed to reveal that a single naturally occurring alanine-to-threonine substitution at position 598, located in the extracellular part of the TM subunit, fully accounted for the lack of activity of the ROD14 Env in HeLa and 12D7 cells. A second mutation at position 422, substituting a lysine residue in ROD10 for an arginine in ROD14, was additionally required for efficient virus release from infected H9 cells, suggesting cell-type specific requirements for this activity. Interestingly, the ROD14 Env protein exhibited a trans-dominant negative effect on particle release by ROD10 Env, suggesting that the viral release activity of the HIV-2 ROD envelope protein may be regulated by its ability to assemble into functional oligomeric structures. PMID- 12726730 TI - Mu-opioid modulation of HIV-1 coreceptor expression and HIV-1 replication. AB - A substantial proportion of HIV-1-infected individuals are intravenous drug users (i.v.DUs) who abuse opiates. Opioids induce a number of immunomodulatory effects that may directly influence HIV-1 disease progression. In the present report, we have investigated the effect of opioids on the expression of the major HIV-1 coreceptors CXCR4 and CCR5. For these studies we have focused on opiates which are ligands for the mu-opioid receptor. Our results show that DAMGO, a selective mu-opioid agonist, increases CXCR4 and CCR5 expression in both CD3(+) lymphoblasts and CD14(+) monocytes three- to fivefold. Furthermore, DAMGO-induced elevation of HIV-1 coreceptor expression translates into enhanced replication of both X4 and R5 viral strains of HIV-1. We have confirmed the role of the mu opioid receptor based on the ability of a mu-opioid receptor-selective antagonist to block the effects of DAMGO. We have also found that morphine enhances CXCR4 and CCR5 expression and subsequently increases both X4 and R5 HIV-1 infection. We suggest that the capacity of mu-opioids to increase HIV-1 coreceptor expression and replication may promote viral binding, trafficking of HIV-1-infected cells, and enhanced disease progression. PMID- 12726731 TI - Only one pRNA hexamer but multiple copies of the DNA-packaging protein gp16 are needed for the motor to package bacterial virus phi29 genomic DNA. AB - A common feature in the maturation of linear dsDNA viruses is that the lengthy viral genome is translocated with remarkable velocity into a limited space within a preformed protein shell using ATP as motor energy. Most biomotors, such as myosin, kinesin, DNA-helicase, and RNA polymerase, contain one ATP-binding component that acts processively. An examination of the well-studied dsDNA viruses reveals that DNA packaging motors involve two nonstructural components. Which component of the motor is the integrated processive factor to turn the motor has not been identified. In bacterial virus phi 29, these two components consist of a gp16 protein and an RNA molecule called pRNA. We have previously predicted and recently confirmed that gp16 binds ATP. It is generally believed that gp16 serves as an ATP-binding and processive component to drive the motor. In this article, phi 29 DNA-packaging intermediates were purified in quantity and examined to differentiate the role between gp16 and pRNA. It was found that the pRNA hexamer is an integral motor component, while gp16 is not stably bound. Only one pRNA hexamer, but multiple copies of gp16, were needed to accomplish DNA packaging. pRNA functions continuously during the entire DNA translocation process, suggesting that pRNA is a vital part of the DNA packaging motor. PMID- 12726732 TI - Identification of genes associated with adenovirus 12 tumorigenesis by microarray. AB - A total of 242 genes were shown to be differentially expressed between haplotypically matched tumorigenic adenovirus 12 (Ad12) and nontumorigenic Ad5 transformed cells using a microarray containing 8734 cDNAs. Eighty-seven of the differentially expressed genes have known roles that include signal transduction, cell growth and proliferation, transcription regulation, protease, and immune functions. The remaining differentially expressed genes are represented by EST cDNAs which have functions that are either completely unknown or proposed, based on sequence similarity to known genes. A subset of 22 differentially expressed genes from the microarray was further examined by Northern blot analyses to verify the identification of new genes associated with Ad12 tumorigenesis. Growth factor receptor binding protein 10 (Grb10) and protease nexin 1 (PN-1) were overexpressed in all of the tumorigenic Ad12-transformed cells examined, whereas expression of these genes was negligible in all of the nontumorigenic Ad5 transformed cells. By contrast, other genes including B cell translocation gene 2 (BTG2) were shown to be significantly up-regulated in Ad5-transformed cells as compared to Ad12-transformed cells. PMID- 12726733 TI - Mapping the active site of vaccinia virus RNA triphosphatase. AB - The RNA triphosphatase component of vaccinia virus mRNA capping enzyme (the product of the viral D1 gene) belongs to a family of metal-dependent phosphohydrolases that includes the RNA triphosphatases of fungi, protozoa, Chlorella virus, and baculoviruses. The family is defined by two glutamate containing motifs (A and C) that form the metal-binding site. Most of the family members resemble the fungal and Chlorella virus enzymes, which have a complex active site located within the hydrophilic interior of a topologically closed eight-stranded beta barrel (the so-called "triphosphate tunnel"). Here we queried whether vaccinia virus capping enzyme is a member of the tunnel subfamily, via mutational mapping of amino acids required for vaccinia triphosphatase activity. We identified four new essential side chains in vaccinia D1 via alanine scanning and illuminated structure-activity relationships by conservative substitutions. Our results, together with previous mutational data, highlight a constellation of six acidic and three basic amino acids that likely compose the vaccinia triphosphatase active site (Glu37, Glu39, Arg77, Lys107, Glu126, Asp159, Lys161, Glu192, and Glu194). These nine essential residues are conserved in all vertebrate and invertebrate poxvirus RNA capping enzymes. We discerned no pattern of clustering of the catalytic residues of the poxvirus triphosphatase that would suggest structural similarity to the tunnel proteins (exclusive of motifs A and C). We infer that the poxvirus triphosphatases are a distinct lineage within the metal-dependent RNA triphosphatase family. Their unique active site, which is completely different from that of the host cell's capping enzyme, recommends the poxvirus RNA triphosphatase as a molecular target for antipoxviral drug discovery. PMID- 12726734 TI - The Potato virus X TGBp3 protein associates with the ER network for virus cell-to cell movement. AB - Potato virus X (PVX) TGBp3 is required for virus cell-to-cell movement. Cell-to cell movement of TGBp3 was studied using biolistic bombardment of plasmids expressing GFP:TGBp3. TGBp3 moves between cells in Nicotiana benthamiana, but requires TGBp1 to move in N. tabacum leaves. In tobacco leaves GFP:TGBp3 accumulated in a pattern resembling the endoplasmic reticulum (ER). To determine if the ER network is important for GFP:TGBp3 and for PVX cell-to-cell movement, a single mutation inhibiting membrane binding of TGBp3 was introduced into GFP:TGBp3 and into PVX. This mutation disrupted movement of GFP:TGBp3 and PVX. Brefeldin A, which disrupts the ER network, also inhibited GFP:TGBp3 movement in both Nicotiana species. Two deletion mutations, that do not affect membrane binding, hindered GFP:TGBp3 and PVX cell-to-cell movement. Plasmids expressing GFP:TGBp2 and GFP:TGBp3 were bombarded to several other PVX hosts and neither protein moved between adjacent cells. In most hosts, TGBp2 or TGBp3 cannot move cell-to-cell. PMID- 12726735 TI - The complete nucleotide sequence, genome organization, and origin of human adenovirus type 11. AB - The complete DNA sequence and transcription map of human adenovirus type 11 are reported here. This is the first published sequence for a subgenera B human adenovirus and demonstrates a genome organization highly similar to those of other human adenoviruses. All of the genes from the early, intermediate, and late regions are present in the expected locations of the genome for a human adenovirus. The genome size is 34,794 bp in length and has a GC content of 48.9%. Sequence alignment with genomes of groups A (Ad12), C (Ad5), D (Ad17), E (Simian adenovirus 25), and F (Ad40) revealed homologies of 64, 54, 68, 75, and 52%, respectively. Detailed genomic analysis demonstrated that Ads 11 and 35 are highly conserved in all areas except the hexon hypervariable regions and fiber. Similarly, comparison of Ad11 with subgroup E SAV25 revealed poor homology between fibers but high homology in proteins encoded by all other areas of the genome. We propose an evolutionary model in which functional viruses can be reconstituted following fiber substitution from one serotype to another. According to this model either the Ad11 genome is a derivative of Ad35, from which the fiber was substituted with Ad7, or the Ad35 genome is the product of a fiber substitution from Ad21 into the Ad11 genome. This model also provides a possible explanation for the origin of group E Ads, which are evolutionarily derived from a group C fiber substitution into a group B genome. PMID- 12726736 TI - Sequences within both the 5' untranslated region and the gag gene are important for efficient encapsidation of Mason-Pfizer monkey virus RNA. AB - It has previously been shown that the 5' untranslated leader region (UTR), including about 495 bp of the gag gene, is sufficient for the efficient encapsidation and propagation of Mason-Pfizer monkey virus (MPMV) based retroviral vectors. In addition, a deletion upstream of the major splice donor, SD, has been shown to adversely affect MPMV RNA packaging. However, the precise sequence requirement for the encapsidation of MPMV genomic RNA within the 5' UTR and gag remains largely unknown. In this study, we have used a systematic deletion analysis of the 5' UTR and gag gene to define the cis-acting sequences responsible for efficient MPMV RNA packaging. Using an in vivo packaging and transduction assay, our results reveal that the MPMV packaging signal is primarily found within the first 30 bp immediately downstream of the primer binding site. However, its function is dependent upon the presence of the last 23 bp of the 5' UTR and approximately the first 100 bp of the gag gene. Thus, sequences that affect MPMV RNA packaging seem to reside both upstream and downstream of the major splice donor with the downstream region responsible for the efficient functioning of the upstream primary packaging determinant. PMID- 12726737 TI - Echinococcosis--an international public health challenge. AB - This review aims to summarise some of the recent studies that have been undertaken on parasites of the genus Echinococcus and the diseases which they cause. Although the adult parasite, which inhabits the intestine of various carnivore species is not pathogenic, the larval or metacestode stages can be highly pathogenic, causing economic losses to livestock and various forms of echinococcosis in humans, some of which have a high fatality rate. There is growing evidence that there are at least 5 species of Echinococcus rather than the generally accepted 4 species. Within these species there are a number of genotypes or strains. This can have implications for surveillance and control. In some wealthy countries, cystic echinococcosis caused by Echinococcus granulosus has been successfully controlled or indeed eradicated. However, in most parts of the world it remains a serious threat to human health. In the former Soviet Union, the disease has rapidly increased in incidence after the end of communist administration. Human alveolar echinococcosis, caused by Echinococcus multilocularis, is more sporadic. However, in some Chinese communities there is a disturbingly high human prevalence and in Europe there has been an increase in the detection rate of E. multilocularis in animals in the last 10 years. Echinococcosis can present diagnostic challenges, particularly in the definitive host in areas of low endemicity. Much of the recent work relating to the use of coproantigen and PCR to overcome these difficulties is summarized. New ideas for controlling the parasite are becoming available and these include both the use of vaccination and the application of mathematical models to determine the most cost effective means of control. Effective measures that are affordable are vital if the parasite is to be controlled in poor countries. PMID- 12726738 TI - Lesions of the pericardium and their significance in the aetiology of heart failure in broiler chickens. AB - The present study focuses on lesions of the pericardium commonly observed in fast growing broilers. These lesions are examined in the context of electrophysiological and functional changes associated with cardiac performance and patho-physiology in broilers succumbing to acute or chronic heart failure. Typical lesions involving the pericardium in fast growing broiler chickens included: (1) excessive pericardial effusion, (2) locally extensive or focal adhesions between parietal and visceral components of the pericardium, (3) fibrous deposits on visceral pericardium, and (4) thickened pericardium. Echocardiographic evidence indicated that severe pericardial effusion and/or adhesions may have a restrictive effect on heart pump function, where both diastolic and systolic function of the heart may be affected. Electrocardiographic data showed a strong trend indicating that pericardial adhesions may be associated with ventricular arrhythmia and increased risk of sudden death in fast growing broilers. Relatively high levels of matrix metalloproteinase MMP-2 activity have been found in pericardial effusions from affected chickens, suggesting a possible involvement of this enzyme in the aetiology of pericardial lesions. The present results indicate that pericardial lesions may be associated with biochemical, morphological, electrophysiological, and functional changes occurring in the hearts of broilers succumbing to acute or chronic heart failure and ascites. PMID- 12726739 TI - Application of touchdown enzyme time release (TETR)-PCR for diagnosis of Chlamydophila abortus infection. AB - Chlamydophila abortus-DNA was detected using a touchdown enzyme time-release (TETR)-polymerase chain reaction (PCR) assay as an improved test for sensitive and rapid diagnosis of abortion in small ruminants. Two hundred and fifty two placentae, liver or spleen tissue samples from aborting ewes and goats or aborted lambs and kids in which C. abortus infection was suspected were examined by TETR PCR and the results were compared with cell culture. Sixty-five tissue samples were found to be TETR-PCR positive while only 56 samples were cell culture positive. After resolution of discrepant samples with a confirmatory nested PCR assay, TETR-PCR had a sensitivity of 97% and a specificity of 99.5% while culture had a sensitivity of 84.8% and a specificity of 100%. The analytical sensitivity of the TETR-PCR assay was determined with DNA extracted from 4-fold serial dilution of C. abortus B577 culture and found to be 0.25 inclusion-forming unit per PCR. No reduction in the analytical sensitivity was noted when the assay was tested with mouse liver samples spiked with 4-fold serial dilution of C. abortus B577 culture. No target product was amplified when DNA from Chlamydophila pecorum was tested. TETR-PCR used in this study is a practical, rapid, sensitive and specific assay that could be used for the detection of C. abortus in infected tissue samples. We recommend the use of this assay as a supplemental diagnostic tool for detection of C. abortus in infected tissue samples. PMID- 12726740 TI - Structural and biochemical evidence of mitochondrial depletion in pigs with hypertrophic cardiomyopathy. AB - Pig hearts with naturally occurring hypertrophic cardiomyopathy (HCM) were isolated to investigate the effects of mitochondrial deficiency at biochemical and molecular levels. Enzyme activities of mitochondrial-encoded cytochrome c oxidase and NADH dehydrogenase in the HCM hearts (n=12) were lower than that in the controls (n=12) by 41+/-29% (P<0.01) and 43+/-21% (P<0.001), respectively. Additionally, Southern blot analysis was conducted to quantify the relative amount of mitochondrial DNA (mtDNA) from the HCM and controls. The relative amount of mtDNA in the HCM hearts was significantly 57+/-19% (P<0.001) lower than that in the controls. Both mitochondrial enzyme deficiency and mtDNA depletion were significantly correlated with the degree of cardiac hypertrophy judged based on the ratio of heart/body weight. In conclusion, our results reveal that a secondary effect of tissue-specific mtDNA depletion and mitochondrial dysfunction is in response to the HCM. PMID- 12726742 TI - Optimal enhancement of in situ hybridization for the detection of porcine circovirus 2 in formalin-fixed, paraffin-wax-embedded tissues using a combined pretreatment of thermocycler and proteinase K. AB - Optimal enhancement of the hybridization signal was developed for the detection of porcine circovirus (PCV) 2 in formalin-fixed, paraffin-wax-embedded tissues from pigs with postweaning multisystemic wasting syndrome. The hybridization signal obtained after thermocycler pretreatment was very uniform across the section, whereas the signal obtained after either proteinase K or microwave pretreatment not only was weaker but was of variable intensity across sections. Thermocycler pretreatment combined with brief proteinase K digestion can enhance signal detection for target viral nucleic acid in formalin-fixed, paraffin-wax embedded tissues. A strong hybridization signal was detected in the cytoplasm of macrophages and multinucleated giant cells in lymph node and spleen without background staining and morphological damage. The technical improvement results, therefore, in an identical background at the same time as an increased signal and, thus, may help detect lower levels of PCV2 DNA in formalin-fixed, paraffin wax-embedded tissues. PMID- 12726741 TI - Cell-specific activation of aflatoxin B1 correlates with presence of some cytochrome P450 enzymes in olfactory and respiratory tissues in horse. AB - Horses may be exposed to aflatoxin B(1) (AFB(1)) via inhalation of mouldy dust, leading to high exposure of olfactory and respiratory tissues. In the present study the metabolic activation of AFB(1) was examined in olfactory and respiratory tissues in horse. The results showed covalent binding of AFB(1) metabolites in sustentacular cells and cells of Bowman's glands in the olfactory mucosa, in some cells of the surface epithelium of nasal respiratory, tracheal, bronchial and bronchiolar mucosa and in some glands in these areas. Immunohistochemistry revealed that cells expressing proteins reacting with CYP 3A4- and CYP 2A6/2B6-antibodies had a similar distribution as those having capacity to activate AFB(1). Our data indicate that the cell-specific activation of AFB(1) correlates with presence of some CYP-enzymes in olfactory and respiratory tissues in horse. PMID- 12726743 TI - Pharmacokinetics of enrofloxacin in lactating sheep. AB - The pharmacokinetics of enrofloxacin (ENR) was investigated after its intravenous (iv) and intramuscular (im) administration in six healthy lactating sheep. After iv ENR injection (as a bolus), the elimination half-life (t(1/2beta)), the volume of distribution (Vd(area)), and the area under the concentration vs. time curve (AUC) were 3.30 (0.36)h, 2.91 (0.17)l/kg and 4.19 (0.18) microg h/ml, respectively. The maximum milk concentrations of ENR (C(max)), the area under the milk concentration vs. time curve (AUC(milk)) and the ratio AUC(milk)/AUC(serum) were 2.38 (0.14)microg/ml, 23.76 (2.21) microg h/ml and 5.62 (0.30), respectively. After im administration of ENR the t(1/2beta), C(max), time of C(max) (t(max)) and absolute bioavailability (F(abs)) were 3.87 (0.10)h, 0.74 (0.07) microg/ml, 0.83 (0.12)h and 75.35%, respectively. The C(max), AUC(milk) and the ratio AUC(milk)/AUC(serum) were 1.94 (0.13) microg/ml, 24.81 (2.25) microg h/ml and 8.15 (0.96), respectively. PMID- 12726744 TI - Pharmacokinetic-pharmacodynamic integration of danofloxacin in the calf. AB - The pharmacokinetics and pharmacodynamics of danofloxacin were studied in calves after intravenous (IV) and intramuscular (IM) administration, at a dose of 1.25 mg/kg in a two period cross-over study, using tissue cages to monitor aspects of extravascular distribution. Danofloxacin had a high volume of distribution (3.90 L/kg) and relatively rapid clearance (1.02 L/kgh) after IV dosing. Terminal half life was 2.65 and 4.03 h, respectively, after IV and IM administration. Danofloxacin penetrated slowly into and was cleared slowly from tissue cage fluid (transudate), elimination half-life (10.2 h after IV and 8.9 h after IM dosing) being greater than for serum. The antibacterial actions of danofloxacin against the pathogen Mannheimia haemolytica 3575 were established in vitro in Mueller Hinton Broth, serum and transudate. These data were used together with in vivo pharmacokinetic parameters, C(max) and AUC to determine the surrogate markers of antimicrobial activity, C(max)/MIC, AUC/MIC and T>MIC. The antibacterial actions of danofloxacin were also determined ex vivo in serum and transudate samples harvested at pre-determined times after IM danofloxacin dosing. Ex vivo AUC/MIC data were integrated with ex vivo bacterial count to establish values producing a bacteriostatic action, inhibition of bacterial count by 50%, reduction in bacterial count by 99.9% (bactericidal action) and elimination of bacteria. Mean values were, respectively, 15.9, 16.7, 18.15 and 33.5h for serum and 15.0, 16.34, 17.8 and 30.7 h for transudate. The AUC/MIC-effect relationships for serum may be regarded as representative of a shallow compartment of blood and well perfused tissues, whilst AUC/MIC-effect relationships for transudate may be considered to represent a deep peripheral compartment of poorly perfused tissues. A novel approach to selecting antimicrobial drug dosage for evaluation in clinical trials, using AUC/MIC values producing either bactericidal activity or elimination of bacteria together with MIC(90) values for calf pathogens, is proposed. This approach can be expected to optimise efficacy and minimise the development of resistance. PMID- 12726745 TI - Putative biomarkers for evaluating antibiotic treatment: an experimental model of porcine Actinobacillus pleuropneumoniae infection. AB - Biomarkers of infection were screened for their possible role as evaluators of antibiotic treatment in an aerosol infection model of porcine pneumonia caused by Actinobacillus pleuropneumoniae (Ap). Following infection of 12 pigs, clinical signs of pneumonia developed within 20 h, whereafter the animals received a single dose of either danofloxacin (2.5mg/kg) or tiamulin (10 mg/kg). To test the discriminative properties of the biomarkers, the dosage regimens were designed with an expected difference in therapeutic efficacy in favour of danofloxacin. Accordingly, the danofloxacin-treated pigs recovered clinically within 24h after treatment, whereas tiamulin-treated animals remained clinically ill until the end of the study, 48 h after treatment. A similar picture was seen for the biomarkers of infection. During the infection period, plasma C-reactive protein (CRP), interleukin-6 and haptoglobin increased, whereas plasma zinc, ascorbic acid and alpha-tocopherol decreased. In the danofloxacin-treated animals, CRP, interleukin 6, zinc, ascorbic acid and alpha-tocopherol reverted significantly towards normalisation within 24h of treatment. In contrast, signs of normalisation were absent (CRP, zinc and ascorbic acid) or less marked (interleukin-6 and alpha tocopherol) in the tiamulin-treated animals. Plasma haptoglobin remained elevated throughout the study in both groups. This indicates that CRP, zinc, ascorbic acid and to a lesser extent interleukin-6 and alpha-tocopherol might be used to evaluate antibiotic treatment of acute Ap-infection in pigs. The present model provides a valuable tool in the evaluation of antibiotic treatments, offering the advantage of clinical and pathological examinations combined with the use of biochemical infection markers. PMID- 12726746 TI - Evaluation of a single dose versus a divided dose regimen of danofloxacin in treatment of Actinobacillus pleuropneumoniae infection in pigs. AB - A single versus a divided dose regimen of danofloxacin was evaluated in treatment of porcine Actinobacillus pleuropneumoniae infection using clinical observations combined with biochemical infection markers: C-reactive protein, zinc and ascorbic acid. Twenty hours after experimental infection, the 18 pigs received danofloxacin intravenously as a single dose of 2.5mg/kg or four doses of 0.6 mg/kg administered at 24h intervals. These dosage regimens resulted in similar AUCs of the plasma danofloxacin vs time curve. The maximum concentration was 3.5 fold higher using the single dose regimen, while the time with concentrations above the MIC was 2.5-fold longer using the fractionated regimen. Using the single dose regimen, temperature was normalised 32 h post-infection. In contrast, normalisation was delayed until 44 h post-infection using four low doses and a relapse with elevated temperatures at 52 and 68 h was observed. No other significant differences between the treatments were found, neither regarding clinical, haematological nor biochemical observations. The use of the more convenient single dose regimen was appropriate, as it was at least equivalent to the fractionated regimen. PMID- 12726747 TI - Free amino-acid concentrations in the equine placenta: relationship to maternal and fetal plasma concentrations. AB - Free amino-acid concentrations were measured in maternal venous and fetal umbilical vein plasma, and in the allantochorion, of Thoroughbred mares at term. Concentrations in maternal and fetal plasma were similar to those reported previously in equids. The concentrations of free amino-acids in the allantochorion were higher than those in the maternal and fetal plasmas and were characterised by high levels of the nonessential amino-acids as observed in other species. Fourteen of the 20 amino-acids measured had similar allantochorion/umbilical vein concentration ratios suggesting that simple gradient diffusion might play a part in their transfer from the placenta to the fetus. PMID- 12726748 TI - Isolation and characterisation of Shiga toxin-producing Escherichia coli O157:H7 from calves in Argentina. AB - Four hundred and twenty-two calves were examined for intestinal carriage of Shiga toxin-producing Escherichia coli O157:H7 using conventional plating. Two (0.5%) E. coli O157 were recovered. They were compared with 96 Argentine strains of different origin by pulsed-field gel electrophoresis, phage typing and PCR-RFLP of stx2 genes. One strain isolated from a calf, was closely related with 18 strains of clinical origin. PMID- 12726749 TI - Patagonian Argentine Creole cattle polymorphism: comparison with North-West populations of this breed. AB - The relict Patagonian Argentine Creole cattle population consist of a small feral population (Los Glaciares population) that is geographically isolated in the South-West of Patagonia. In order to determine the level of genetic variability of this population, the polymorphism of eight structural genes and two microsatellites loci were studied using the polymerase chain reaction (PCR). In addition, genetic characterisation was used to compare Los Glaciares population and the ACc breed of cattle. Results obtained in this study show that the value of average heterozygosity of the studied loci for the Los Glaciares were not significantly different from the ACc. Furthermore, the data of this report were consistent with the hypothesis that Los Glaciares originated from ACc brought to the area by colonialists in the last century. Such data may be useful in formulating management plans for Feral Patagonian Creole cattle populations. PMID- 12726750 TI - The limits of science: dioxin. PMID- 12726752 TI - Respiratory cancer risks associated with low-level nickel exposure: an integrated assessment based on animal, epidemiological, and mechanistic data. AB - Increased lung and nasal cancer risks have been reported in several cohorts of nickel refinery workers, but in more than 90% of the nickel-exposed workers that have been studied there is little, if any evidence of excess risk. This investigation utilizes human exposure measurements, animal data from cancer bioassays of three nickel compounds, and a mechanistic theory of nickel carcinogenesis to reconcile the disparities in lung cancer risk among nickel exposed workers. Animal data and mechanistic theory suggest that the apparent absence of risk in workers with low nickel exposures is due to threshold-like responses in lung tumor incidence (oxidic nickel), tumor promotion (soluble nickel), and genetic damage (sulfidic nickel). When animal-based lung cancer dose response functions for these compounds are extrapolated to humans, taking into account interspecies differences in deposition and clearance, differences in particle size distributions, and human work activity patterns, the predicted risks at occupational exposures are remarkably similar to those observed in nickel-exposed workers. This provides support for using the animal-based dose response functions to estimate occupational exposure limits, which are found to be comparable to those in current use. PMID- 12726753 TI - The effects of diisopropylmethylphosphonate, a by-product of the production of sarin and a contaminant in drinking water at the Rocky Mountain Arsenal, on female mink. AB - This paper challenges the interpretation of the report on the effects of DIMP on mink with respect to mortality causation and organ-specific toxicity (i.e., liver, kidney, and thymus). During the second generation (F(1)) of a two generation toxicity study by on the effects of DIMP on female brown Ranch Wild mink, a cluster of six premature deaths occurred, being attributed to an unintentional anesthetic overdose that induced a stress-related syndrome rather than DIMP exposure. The present paper reveals that the six adult female mink (i.e., four DIMP treated and two controls) had medical conditions that pre existed the administration of anesthetic. Three of the four DIMP-exposed mink that died displayed evidence of medically significant red blood cell damage and immune system involvement, while all six mink had elevated blood serum enzymes (ALT/AST) indicative of liver damage along with confirmatory histopathology indicating severe liver damage. These findings challenge the conclusion of Bucci et al. that the six mink deaths were solely caused by stress induced by anesthesia, and suggest DIMP related pre-existing conditions (e.g., red blood cell damage and elevated ALT/AST) may have contributed to the deaths in DIMP treated mink. also inexplicably combined both the pregnant and non-pregnant animal data, thereby precluding an assessment of the effects of DIMP on pregnant and non-pregnant mink. A re-evaluation of the findings of the data revealed that pregnancy/lactation significantly influenced the incidence of physiologic alterations and histological lesions in the female mink for the kidney, liver, and thymus, findings that were masked by the combining of pregnant and non pregnant results. Further evaluations indicated that DIMP treatment also significantly induced liver lesions in pregnant mink and kidney lesions in both pregnant and non-pregnant mink. These findings challenge the conclusions of Bucci et al. concerning the effects of DIMP on mink as well as identify for the first time that pregnancy/lactation in mink is a risk factor in the induction of thymic atrophy and the elevation of serum ALT/AST. PMID- 12726754 TI - Dioxin risks in perspective: past, present, and future. AB - The United States Environmental Protection Agency (USEPA) and other U.S. and international agencies have focused extensive efforts on the evaluation of the potential health risks of exposures to chlorinated dioxins (PCDDs), furans (PCDFs), and related dioxin-like polychlorinated biphenyls (PCBs). Extensive regulatory efforts over the past 20 years have also been made to control emissions of these compounds and thus to reduce exposures in the general population. This paper reviews the available information on temporal trends in emissions, environmental levels, intake levels through foods, and human body burdens of dioxins. This paper also provides an overview and comparison of recent hazard assessments for dioxins from U.S. and international agencies. Available data on emissions, environmental and food levels, and human body burdens of dioxins in the general population indicate a several-fold reduction in exposures and body burdens in the general population over the three decades from 1970 to 2000. U.S. and international hazard assessments concur on certain aspects, but disagree on fundamental issues including the likelihood of a threshold for carcinogenic dose-response and the degree of safety factors needed in deriving a protective exposure limit. These disagreements have significant consequences for interpreting the potential health risks of current background dioxin exposure levels. However, whatever the degree of health risk that may be associated with current background exposures, the general population is experiencing several-fold lower exposures, and, therefore, lower health risks, currently compared to 30 years ago. In light of the dramatic declines in exposure already observed, further efforts to reduce exposures through attempts to control emissions or food levels should be carefully evaluated to understand the likely efficacy of the efforts and the relative costs and benefits. PMID- 12726755 TI - The safety assessment of fragrance materials. AB - Safety evaluation of the large number of diverse chemicals used as fragrance ingredients follows a systematic prioritization of data generation and analysis, consideration of exposure and critical analysis of the quality of the available information. In prior publications the research priorities used by the Research Institute for Fragrance Materials (RIFM), and the methods of exposure estimation used by industry have been summarized. This paper provides details of the approach used by the RIFM Expert Panel (REXPAN), to examine the dermal effects, systemic toxicity and environmental consequences of the use of and exposure to fragrance materials, which allow a reliable determination of safe use under intended conditions. The key to the usefulness of this analysis is the grouping of more than 2600 discrete ingredients into classes, based on chemical structures. Research sponsored by RIFM, data supplied by member companies, and relevant published reports from many sources are all considered during hazard characterization. A discussion is provided of REXPAN's decision tree approach to assessing the dermal, systemic and environmental endpoints and the types and quality of data included. This overall process results in well-documented conclusions which are provided to the International Fragrance Association (IFRA) as the basis for consideration of a new or existing Fragrance Material Standard and to industry for appropriate product risk management actions. PMID- 12726756 TI - Summary of safety studies conducted with synthetic lycopene. AB - Lycopene belongs to the group of natural carotenoids, which are found in many fruits and vegetables, but predominantly in tomatoes and tomato-based products. This manuscript summarizes the safety of synthetic lycopene as a water dispersible beadlet formulation containing antioxidants and includes acute and subchronic safety studies, reproductive studies, genotoxicity studies, metabolic studies, and exploratory studies on the hepatic uptake of lycopene. Lycopene has a low order of acute toxicity and no significant toxicity has been observed in rats treated with lycopene beadlet formulations in the diet at doses of up to 500 mg/kg bw/day for 14 weeks or 1000 mg/kg bw/day for 4 weeks. No teratogenic effects were noted in a rat two-generation study (1000 ppm in the diet) or in a teratology study in rats with 1000 mg/kg bw/day lycopene as beadlet formulations. Lycopene accumulates in hepatocytes and to a lesser extent in spleen. In short term studies with synthetic lycopene, as a beadlet formulation, and natural source lycopene, as tomato concentrate, the accumulation of lycopene in the liver and the presence of pigment deposits in the hepatocytes were similar and neither was associated with any histopathological changes. The pigment deposits in hepatocytes are no longer present after approximately 13 weeks of depletion, demonstrating reversibility for this effect. Unformulated pure crystalline lycopene and lycopene as a 10% beadlet formulation are not genotoxic as determined in a comprehensive battery of tests, however, improperly stored, unformulated crystalline lycopene can degrade to mutagenic products if exposed to light and air. Lycopene is commercially available only in formulated forms, containing antioxidants, which prevent the degradation of lycopene and other excipients that provide for water dispersibility. In the animal studies, there is a large margin of safety based on the repeated dose safety and reproductive/teratology studies in rodents. In humans, there is a very long history of use with respect to dietary exposure, and even in the case of very high exposures from dietary sources, there is no indication of any significant adverse effects. PMID- 12726757 TI - Safety evaluation of a phytase, expressed in Schizosaccharomyces pombe, intended for use in animal feed. AB - BD006 phytase is the product of the Escherichia coli B app A gene expressed in Schizosaccharomyces pombe strain ASP595-1. This enzyme preparation is intended for use in animal feed applications where it improves the bioavailability of phosphate for monogastric animals. BD006 phytase was tested as an unrefined (DV006U) and a refined (DV006R) preparation for its effects on genotoxicity and in acute, inhalation and subchronic toxicity studies. Dosages ranged from 5000 microg/plate for in vitro toxicity studies to a high of 2000X for oral in vivo toxicity studies. The highest oral dose tested (2000X) is 2000 times the highest expected consumption of product by poultry or swine (X=50 units of phytase per kg bwt/day). There was no genotoxicity or any in vivo toxicity reported which could be directly related to systemic effects of the product. Other additional safety studies conducted were devoid of any relevant toxicity. The historic safety profile of the production organism S. pombe, and the results of the toxicity studies presented herein, attest to the safety of BD006 phytase for use in animal feed applications. PMID- 12726758 TI - Safety evaluation of lipase produced from Rhizopus oryzae: summary of toxicological data. AB - The toxicity of Lipase D, an enzyme preparation, was evaluated in a series of studies. Lipase D selectively hydrolyzes triglycerides of fatty acids. It also catalyzes the interesterification of edible fats and oils. In a 13-week gavage study, Sprague-Dawley rats received Lipase D at levels of 0, 500, 1000, or 2000 mg/kg body wt./day. A dose dependent decrease in urinary pH was observed, but there were no effects on electrolyte balance, kidney weight, or histology of the kidney. The no-observed-adverse-effect level in rats was 1000 mg/kg body wt./day. In common with other enzyme preparations, Lipase D was not genotoxic. Lipase D was tested in the Ames assay, the mouse lymphoma forward mutation assay, and the chromosome aberration assay. Finally, the particular strain of Rhizopus oryzae used to prepare Lipase D was shown to have low to moderate pathogenicity when injected into the tail vein of mice at doses up to 1.3 x 10(6) colony-forming units (CFU) per animal. No effects were observed when mice received up to 2.2 x 10(5) CFU by gavage or in their diets daily for 28 days. The results indicate that this particular strain can be handled using ordinary safety practices current in the fermentation industry. These studies support a conclusion that Lipase D is safe when used as described in the processing of dietary fatty acids and glycerides of fatty acids. PMID- 12726759 TI - Utilizing data from multiple studies (meta-analysis) to determine effective dose duration levels. Example: rats and mice exposed to hydrogen sulfide. AB - The objective of this exercise was to incorporate as much data as possible from multiple studies, that may differ in exposure durations, to derive a chemical specific dose-duration response curve from which to identify toxicity markers (e.g., ED01, benchmark dose, and LD50). This has the advantage of incorporating more information than single-study assessments to improve estimates and reduce confidence intervals, and determining toxicity markers as functions of exposure duration as well as dose. The example used mortality for rats and mice, analyzed separately, from acute exposure to hydrogen sulfide (dose refers to airborne concentration of H(2)S). Statistical methods were applied to determine when data from different studies could be pooled. EC01, EC10, and EC50 (doses with response rates of 1, 10, and 50%) were estimated, with 95% confidence intervals, at durations of 5, 10, and 30 min, and 1, 2, 4, and 6 h. A single dose-duration response curve for mortality was fit to the rat data for exposures of 5 min, 10 min, 30 min, and 1h, using a logistic curve additive in log(dose) and log(duration). Separate fits of that model were required, however, at 2, 4, and 6h, due to an increasing impact of duration relative to concentration as duration increased. The curves for rats fit the data exceedingly well and exhibited a threshold-like response followed by a steep incline as concentration increased. There were fewer data for mice but the response pattern for mortality clearly differed from rats. This example demonstrates the feasibility of extending the concept of single-study benchmark doses to multiple-study dose-duration benchmarks, using U.S. EPA's program CatReg. Similar applications to long-term animal studies could be considered. PMID- 12726760 TI - Immunochemical analysis of cytochrome P450 variability in human leukapheresed samples and its consequences for the risk assessment process. AB - Xenobiotic metabolizing cytochrome P450 (P450) enzymes were investigated in leukapheresed samples from 50 human individuals. It was the aim of the study (a). to get insight into the extent of extrahepatic P450 variability, (b). to investigate whether and to which extent P450 expression and variability as it is seen in the liver corresponds to P450 expression at extrahepatic sites, and (c). to contribute to the replacement of traditionally used default factors (usually 10 for interindividual variability) by data-derived factors in the risk assessment process. P450 enzymes were determined by Western Blotting. Immunoquantification was performed for P450 1A, 1B1, 2C, 2D6, 2E1, and 3A which were-with the exception of the polymorphically expressed CYP2D6-detectable in all samples investigated. Amounts of P450 enzymes in leukapheresed samples were (except CYP1B1) lower compared to those reported for the liver. The P450 variabilities were expressed by the ratios between the 95th and the 5th percentiles. They displayed 7-(CYP1A), 4-(CYP1B1), 6-(CYP2C), 30-(CYP2D6), 3 (CYP2E1), and 4-(CYP3A) fold variability in specific protein content. The results show (a). qualitative and quantitative differences in the expression of P450 proteins in leukapheresed samples from 50 individuals compared to liver, (b). a different extent of variability depending on the P450 enzyme, and (c). in cases where polymorphically distributed P450 enzymes are involved, the traditionally used factor of 10 might be too low to account for interindividual variability in both toxicokinetics and toxicodynamics. PMID- 12726761 TI - Use of ADME studies to confirm the safety of epsilon-polylysine as a preservative in food. AB - Epsilon-polylysine is a homopolymer of L-lysine, containing approximately 30 L lysine subunits, as synthesized in aerobic bacterial fermentation by Streptomyces albulus. epsilon -Polylysine is approved for food use in Japan as an antimicrobial preservative. A series of pharmacokinetic and metabolic profile studies on epsilon -polylysine have been conducted in rats in order to provide a better understanding of the reason for its lack of toxicological effects in subchronic and chronic feeding bioassays using relatively high concentrations in the diet up to 50000 ppm. As reported in this article, epsilon -polylysine was practically non-toxic in an acute oral toxicity study in rats, with no mortality up to 5 g/kg and was not mutagenic in bacterial reversion assays. Absorption, distribution, metabolism and excretion (ADME) studies on 14C-radiolabeled epsilon -polylysine, given in a single dose to fasted male rats at 100mg/kg, revealed low absorption from the gastrointestinal tract. All but trace amounts of the dosed radioactivity was eliminated by excretion within 168 h and over 97% was accounted for in urine (1.2%), feces (92.9%), or expired air (3%) by 48 h. The sum of the cumulative excretion with routes associated with absorption in urine, expired air and carcass was 6.4% of total recovered radioactivity; approximately 94% of the dose of epsilon -polylysine passed unabsorbed through the gastrointestinal tract in the feces. Whole body autoradiography did not show concentration of absorbed epsilon -polylysine in any tissue or organ. Excretion half-lives of epsilon polylysine equivalents in blood and plasma were 20 and 3.9 days, likely prolonged by the incorporation into protein of cleaved L-lysine. Metabolic profiles by HPLC analysis of plasma samples suggest that L-lysine is the predominant early metabolic by-product, likely from protease activity in the upper GI tract; only 0.2% of the administered parent compound was found in plasma. At 8-72 h, HPLC profiles show diminishing levels of epsilon -polylysine and L-lysine in plasma, accompanied by a shift to larger peaks of homopolymer fragments of varying subunit length, presumably from microbial degradation of epsilon -polylysine in the lower gut. HPLC profiles of urine and feces collected from 0 to 24 h post dosing revealed three distinct peaks in urine, the first peak likely to be epsilon -polylysine and epsilon -polylysine less a few amino acid subunits, and the second, L-lysine and the third, a metabolite of L-lysine. Radiolabeled L lysine was reduced from 67.2% of the radioactivity in plasma at 30 min to 7.5% at 4 h, indicating that L-lysine is readily removed from plasma from essential amino acid incorporation into protein. Based on the findings of the ADME studies and lack of toxicity in safety studies, the proposed use of epsilon -polylysine as a preservative in foods is considered to be safe. PMID- 12726763 TI - Mobile genetic elements and bacterial toxinoses: the superantigen-encoding pathogenicity islands of Staphylococcus aureus. AB - It is a remarkable observation that virtually all bacterial toxins associated with specific clinical conditions (toxinoses) are encoded by mobile (and therefore variable) genetic elements. Remarkably, these rarely, if ever, carry determinants of antibiotic resistance. Examples are the toxins responsible for diphtheria, anthrax, tetanus, botulism, cholera, toxic shock, scarlet fever, exfoliative dermatitis, food poisoning, travelers' diarrhea, shigella dysentery, necrotizing pneumonia, and others. A recently discovered example of this phenomenon is the family of related staphylococcal pathogenicity islands encoding superantigens (SAgs). These are 15-20kb elements that occupy constant positions in the chromosomes of toxigenic strains, and are characterized by certain phage related features, namely genes encoding integrases, helicases, and terminases, and the presence of flanking direct repeats. The prototype, SaPI1 of Staphylococcus aureus, encodes TSST-1 plus two newly described SAgs, SEK and SEL. Other members of the family encode enterotoxins B (SaPI3) and C (SaPI4), plus at least two other SAgs each. SaPI1 and SaPI2, also encoding TSST-1, are excised and induced to replicate by certain staphylococcal phages, and are then encapsidated at high efficiency into phage-like infectious particles with heads about 1/3 the size of the helper phage heads, commensurate with the sizes of the respective genomes. This results in transfer frequencies of the order of 10(8)/ml, and is presumably responsible for the spread of these elements as well as for their acquisition in the first place. In the absence of a helper phage, these two islands are highly stable; neither excision, loss, or transfer occurs at detectable frequency. Several general implications of this phenomenon will be discussed. One is that the determinants of these toxins have been imported from other species and therefore are not components of the basic genome of the extant producing organisms. This raises the question of the biological (adaptive?) roles of these toxins. Another is that the toxin-carrying units can spread among different (though probably related) species. An interesting question is that of the biological basis for the separation of toxin and resistance determinants. PMID- 12726764 TI - RpoN (sigma(54)) is required for plasmid-encoded coronatine biosynthesis in Pseudomonas syringae. AB - The plant pathogen Pseudomonas syringae pv. glycinea PG4180 produces coronatine (COR), a phytotoxin which functions as a virulence factor in bacterial blight of soybeans. The COR biosynthetic gene cluster in PG4180 is borne on a 90-kb plasmid named p4180A. Although pathway-specific regulatory genes for COR have been identified, global regulatory genes for COR production in PG4180 remain undefined. In the present study, we evaluated the role of rpoN, which encodes sigma(54), in the virulence of strain PG4180. A rpoN mutant of PG4180, designated PG4180.K2, was unable to grow in M9 minimal medium; however, the addition of exogenous glutamate, glutamine or aspartate to M9 medium enabled PG4180.K2 to grow in vitro. PG4180.K2 could not induce disease symptoms or multiply in soybean plants and was defective in COR production and cor gene expression. Furthermore, PG4180.K2 was impaired in transcription of hrpL, an alternate sigma factor that mediates expression of genes in the type III secretion system of P. syringae. PG4180.K2 transconjugants with a wild-type copy of rpoN were complemented for hrpL and cor gene expression, COR biosynthesis, and growth in vitro. Our results indicate that rpoN is required for growth and the expression of both chromosomal and plasmid-encoded virulence factors in P. syringae pv. glycinea PG4180. PMID- 12726765 TI - Sequence and analysis of pBM02, a novel RCR cryptic plasmid from Lactococcus lactis subsp cremoris P8-2-47. AB - This paper reports the complete nucleotide sequence of the 3.85 kbp plasmid pBM02 from Lactococcus lactis subsp. cremoris P8-2-47. Analysis of the sequence predicted six ORFs larger than 25 amino acids. They all were transcribed from the same strand and organized in two functional cassettes: the replication region and a putative mobilization region. In the replication region, two ORFs specifying proteins homologous to others found in some classes of rolling circle-replicating plasmids were encountered (copG and repB). In fact, single-stranded DNA was detected as a replication intermediate of pBM02. copG and repB, together with some upstream sequences, formed part of the minimal replication unit of the plasmid. Interestingly, pBM02 shared a 212 bp stretch with plasmids of the pWV01 type, in which the whole single-strand origin of replication is included. In the mobilization region, an ORF coding for a mobilization-like protein was present, preceded by a putative oriT sequence homologous to that of plasmid pMV158. The replicon of pBM02 is of the wide-host range type, and functions in both Gram positive and Gram-negative bacteria, including Lactobacillus casei, Lactobacillus plantarum, Bacillus subtilis, and Escherichia coli. PMID- 12726767 TI - Chloramphenicol resistance transposable element TnSs1 of Streptococcus suis, a transposon flanked by IS6-family elements. AB - A new transposon, designated TnSs1, which contains a chloramphenicol acetyltransferase gene flanked by direct repeats of an IS6-family element was found in a field isolate of Streptococcus suis. Polymerase chain reaction and hybridization analyses indicated that another field isolate carried the same transposon in a different location on the chromosome. A transposition assay done with a thermosensitive suicide vector showed that, among the seven TnSs1 mutants tested in this study, six formed a cointegrate between the S. suis genome and the vector with the generation of the third copy of the insertion sequence element, and one harbored one copy of TnSs1 on the chromosome as a result of a subsequent resolution step. On transposition, TnSs1 duplicated an 8-bp sequence at the target site. PMID- 12726766 TI - Molecular organization of exopolysaccharide (EPS) encoding genes on the lactococcal bacteriophage adsorption blocking plasmid, pCI658. AB - The lactococcal plasmid pCI658 (58 kb) isolated from Lactococcus lactis ssp. cremoris HO2 encodes the production of a hydrophilic exopolysaccharide (EPS) which consists primarily of galactose and glucuronic acid and which interferes with adsorption of phages o712 and oc2 to cell surface receptors. Examination of the nucleotide sequence of a 21.8-kb region of the plasmid revealed a large genetic cluster consisting of at least 23 putative EPS biosynthetic determinants in addition to the presence of insertion sequences at the 5(') and 3(') ends. According to homology searches, the genes were organized in specific regions involved in regulation, synthesis and export of the EPS. The predicted products of individual genes exhibited significant homology to exopolysaccharide, capsular polysaccharide (CPS), and lipopolysaccharide (LPS) gene products from a variety of Gram positive and Gram negative bacteria. Evidence of a gene encoding UDP glucose dehydrogenase is also presented and this is the first description of such a gene in Lactococcus. PMID- 12726768 TI - Analysis of pCU1 replication origins: dependence of oriS on the plasmid-encoded replication initiation protein RepA. AB - The broad-host-range replicon of the plasmid pCU1 has three origins of vegetative replication called oriB, oriS, and oriV. In the multi-origin replicon, individual origins can distinguish among replication factors provided by the host. It has been found that during replication in Escherichia coli polA(-) host, oriS was the only active origin of a mutant pCU1 derivative bearing a mutation in the gene encoding replication initiation protein RepA. To further investigate the capacity of oriS to function in an E. coli polA(-) host we constructed a number of clones of the basic replicon of pCU1 containing oriS as the only replication origin. An oriS construct created with pUC18 could transform the polA(-) strain when RepA was supplied in trans. When the oriS region (between nucleotides 290 and 832) was ligated to an antibiotic resistance Omega fragment, the construct could be recovered as a plasmid from polA(+) strain if functional RepA was provided in trans. Our results therefore indicate that the basic replicon of pCU1, containing oriS as the sole origin, does require RepA to initiate plasmid replication in E. coli PMID- 12726769 TI - Complete nucleotide sequence of a native plasmid from Brevibacterium linens. AB - Brevibacterium linens has commercial significance in the dairy industry and potential application in the production of bacteriocins and carotenoids. Strain development of these industrially significant organisms would be facilitated by the use of vectors, yet few are available. In this study we report the isolation of four novel plasmids from the Gram-positive coryneform B. linens, and determine the first complete nucleotide sequence of a native plasmid of B. linens. The cryptic plasmid pLIM is 7610 bp in length, and belongs to a subfamily of theta replicating ColE2-related plasmids. Initial investigation suggests that replication in pLIM requires two replicases, a primase (RepA) and a DNA binding protein (RepB), encoded by a single operon repAB. The origin of replication is located upstream of repAB transcription. PMID- 12726770 TI - Characterization of class 1 integron resistance gene cassettes and the identification of a novel IS-like element in Acinetobacter baumannii. AB - Based on hybridization studies, 21/32 multi-resistant clinical isolates of Acinetobacter baumannii contain class 1 integrons. Amplification products were obtained from 20 of the hybridization-positive strains. A single dfrA7 cassette was identified in 18 of the isolates and an integron with two cassettes (aadB aadA4) was found in only one strain. Amplicons were not obtained from one of the hybridization positive strains. DNA sequence analysis of a 6.080-kb fragment, cloned from this strain, identified the remnant of an integron, following insertion of IS26 into the 5(')-end of intI1. The 6.080-kb sequence carries an aminoglycoside resistance gene, linked to a portion of IS1133, which in turn is linked to a sequence that has properties of IS elements, including sequences that could stimulate transcription, and ORFs encoding amino acid sequences with similarity to a transposase from Deinococcus radiodurans. PMID- 12726771 TI - New donor vector for generation of histidine-tagged fusion proteins using the Gateway Cloning System. AB - An optimized donor/shuttle vector, pENTR-His-ccdB, was generated that readily produces a histidine-tagged recombinant protein in multiple expression systems using Gateway Technology. In the current Gateway System, six histidines and the tobacco etch virus protease cleavage site are encoded upstream of the attR1 recombination site such that the histidine-tagged destination/expression vector adds 15 residues to the amino-terminus of recombinant proteins. Our new vector introduces the histidine tag at the donor level and places the multiple cloning sites within the attL recombination sites producing cleavable histidine-tagged proteins with a short, neutral linker of five residues. Two histidine-tagged clones were produced and fusion proteins expressed using the newly engineered vector. PMID- 12726772 TI - Development of a novel bacterial artificial chromosome cloning system for functional studies. AB - Bacterial artificial chromosome (BAC) cloning systems currently in use generate high quality genomic libraries for gene mapping, identification, and sequencing. However, the most commonly used BAC cloning systems do not facilitate functional studies in eukaryotic cells. To overcome this limitation, we have developed pEBAC190G, a new BAC vector that combines the features of the first generation PAC/BAC vectors with eukaryotic elements that facilitate the transfection, episomal maintenance, and functional analysis of large genomic fragments in eukaryotic cells. A number of different cloning strategies may be used to retrofit genomic fragments from existing libraries into the new vector. The system was tested by the retrofitting of a 170kb NotI genomic fragment from the RPCI-11 BAC library into the NotI site of pEBAC190G. Clones from any eukaryotic genomic library harboured in this vector can be transferred from bacteria directly to eukaryotic cells for functional analysis. PMID- 12726773 TI - Identification and nucleotide sequence analysis of a cryptic plasmid, pRM21, from Rhodothermus marinus. AB - Here we report the identification and nucleotide sequence analysis of pRM21, a plasmid isolated from the thermophilic eubacterium Rhodothermus marinus. pRM21 consists of 2935 bp, has a G+C content of 58.2% and one major open reading frame whose deduced product shows significant similarities to RepA proteins from several plasmids, the highest being to the RepA of pSa from Escherichia coli. A region with the characteristics of iteron-containing replicons, three 19 bp repeats, DnaA boxes, an A+T rich region and GATC sequences, was identified. Of 40 additional R. marinus strains screened for plasmids, six (15%) were found to harbour plasmids with the same size and restriction pattern as pRM21. PMID- 12726774 TI - Impact of aging on DNA methylation. AB - The biochemistry of aging is complex, with biologically significant changes occurring in proteins, lipids and nucleic acids. One of these changes is in the methylation of DNA. DNA methylation is a mechanism modifying gene expression. The methylation of sequences in or near regulatory elements can suppress gene expression through effects on DNA binding proteins and chromatin structure. Both increases and decreases in methylation occur with aging, depending on the tissue and the gene. These changes can have pathologic consequences, contributing to the development of malignancies and autoimmunity with aging, and possibly to other disorders as well. Thus, while aging can impact on DNA methylation, the changes in DNA methylation can also impact on aging. This review summarizes current evidence for changes in the methylation status of specific genes with aging, their impact on diseases that develop with aging, and mechanisms that may contribute to the altered DNA methylation patterns. As this field is still developing, it is anticipated that new knowledge will continue to accumulate rapidly. PMID- 12726775 TI - Aging as war between chemical and biochemical processes: protein methylation and the recognition of age-damaged proteins for repair. AB - Deamidated, isomerized, and racemized aspartyl and asparaginyl residues represent a significant part of the spontaneous damage to proteins that results from the aging process. The accumulation of these altered residues can lead to the loss of protein function and the consequent loss of cellular function. However, almost all cells in nature contain a methyltransferase that can recognize the major damaged form of the L-isoaspartyl residue, and some of these enzymes can also recognize the racemized D-aspartyl residue. The methyl esterification reaction can initiate the conversion of these altered residues to the normal L-aspartyl form, although there is no evidence yet that the L-asparaginyl form can be regenerated. This enzyme, the protein L-isoaspartate (D-aspartate) O methyltransferase (EC 2.1.1.77), thus functions as a protein repair enzyme. The importance of this enzyme in attenuating age-related protein damage can be seen by the phenotypes of organisms where the gene encoding has been disrupted, or where its expression has been augmented. PMID- 12726776 TI - Hyper nuclear acetylation (HNA) in proliferation, differentiation and apoptosis. AB - Coactivators such as cyclic AMP-response-element binding protein (CREB)-binding protein (CBP) and p300/CBP associated factor (P/CAF) play a crucial role in coordinating and cointegrating eukaryotic transcription. One of the recent paradigms in the eukaryotic transcription field is the finding of molecular basis of coactivator function. The well characterized coactivators such as CBP and P/CAF have been proposed to coactivate/cointegrate gene expression with many transcription activators through two mechanisms. One is complex formation with the components with basal transcriptional machinery. Another is its intrinsic and associated enzymatic activity, which transfers an acetyl-base to the epsilon ( epsilon ) portion of lysine-residues in histones and certain nuclear proteins (factor acetyltransferases; FATs), such as p53, lymphoid enhancer-binding factor (LEF), and transcription factor IIE (TFIIE), which often results in increased transcriptional activity. Recently, the status of hyper nuclear acetylation (HNA) has been thought to influence proliferation, differentiation and apoptosis. Furthermore, recent reports showed that histone acetyltransferase (HAT) activity is increased in human disease, such as cancer and atherosclerosis, and studies have shown associations between nuclear acetylation/deacetylation and cell proliferation/differentiation. PMID- 12726777 TI - Targeting DNA methylation in cancer. AB - There is overwhelming evidence that DNA methylation patterns are altered in cancer. Methylation of CG-rich islands in regulatory regions of genes marks them for transcriptional silencing. Multiple genes, which confer selective advantage upon cancer cells such as tumor suppressors, adhesion molecules, inhibitors of angiogenesis and repair enzymes are silenced. In parallel, tumor cell genomes are globally less methylated than their normal counterparts. In contrast to regional hypermethylation, this loss of methylation in cancer cells occurs in sparsely distributed CG sequences. We now understand that DNA methylation machineries might include a number of DNA methyltransferases, proteins that direct DNA methyltransferases to specific promoters, chromatin modifying enzymes as well as demethylases. There is also data to suggest that pharmacological down regulation of some members of the DNA methylation machinery could inhibit cancer in vitro, in vivo and in clinical trials. Understanding which functions of DNA methylation machinery are critical for cancer is essential for the design of inhibitors of the DNA methylation machinery as anticancer agents. This review discusses the possible role of DNA methyltranferases and demethylases in tumorigenesis and the possible pharmacological and therapeutic implications of the DNA methylation machinery. PMID- 12726778 TI - Methylation and acetylation in nervous system development and neurodegenerative disorders. AB - The cytoarchitecture and cellular signaling mechanisms of the nervous system are complex, and this complexity is reflected at the molecular level with more genes being expressed in the nervous system than in any other tissue. Gene expression and protein function in neural cells can be regulated by methylation and acetylation. Studies of mice deficient in enzymes that control DNA methylation and of animals with a dietary deficiency of folate have established critical roles for methylation in development of the nervous system. Various neuronal proteins including histones and tubulin are regulated by acetylation which appears to serve important functions in the development, stability and plasticity of neuronal networks. Some inherited neurological disorders have recently been linked to mutations in genes that regulate DNA methylation, and alterations in DNA and protein methylation and/or acetylation have been documented in studies of age-related neurodegenerative disorders including Alzheimer's disease (AD), Parkinson's disease (PD) and Huntington's disease (HD). Manipulations of methylation and acetylation can affect the vulnerability of neurons to degeneration and apoptosis in experimental models of neurodegenerative disorders, suggesting a contribution to altered methylation and acetylation to the disease processes. Interestingly, dietary factors that influence DNA methylation may affect the risk of neurodegenerative disorders, for example, individuals with low dietary folate intake are at increased risk of Alzheimer's and Parkinson's diseases. PMID- 12726779 TI - Employed adolescents and beliefs about self-efficacy to avoid smoking. AB - This paper examines self-efficacy to avoid cigarette smoking and its association with smoking and quitting behavior, peer and worksite influences, nicotine dependence, and socio-demographic variables among employed adolescents. A cross sectional survey was used to collect data from employed adolescents ages 15-18 who worked in 10 participating grocery stores in Massachusetts. Eighty-three percent of workers (n=379) completed the survey. Results from the multivariate model indicate that daily smokers were less confident in their ability to avoid smoking than those who smoked less frequently. As nicotine dependence increased, self-efficacy beliefs decreased. In addition, as friends' encouragement to quit increased, self-efficacy beliefs also increased. Work-related variables were not associated with self-efficacy beliefs among smokers. This study suggests that smoking frequency, nicotine dependence, and friends' encouragement to quit are associated with self-efficacy to avoid smoking. Researchers may tailor interventions for daily and less-than-daily smokers, build on peer networks that encourage quitting and help smokers resist pressures to smoke, and enhance strategies for coping with nicotine dependence in high-risk situations. PMID- 12726780 TI - Individual and social/environmental predictors of alcohol and drug use 2 years following substance abuse treatment. AB - The purpose of this study was to identify individual and social/environmental predictors of alcohol and drug use 2 years following substance abuse treatment. Participants (n=180) self-administered questionnaires within their first month of substance abuse treatment and completed a 2-year follow-up interview. Individual factors (coping, self-efficacy, resource needs, and expectations for sober fun), social/environmental factors (craving, exposure, negative social influences, and involvement in substance-using leisure activities), and background characteristics measured during treatment were used to predict alcohol and drug use during a 2-year follow-up using manifest variable regression analysis. Results suggest that poorer self-efficacy, greater involvement in substance-using leisure activities, being single, and less income predicted alcohol use directly, whereas greater resource needs and involvement in substance-using leisure activities, being of minority status, and being single predicted drug use directly. Income, gender, problem severity, marital status, and race also predicted alcohol and drug use indirectly. Findings highlight differential predictors of posttreatment substance use that may be useful in developing alternative approaches to prevent relapse. PMID- 12726781 TI - Natural reduction of binge drinking among college students. AB - Considerable evidence indicates that alcohol problems can resolve without formal treatment [Addiction 95 (2000) Clin. Psychol.: Sci. Pract. 5 (1998) 1]. Such changes, called "natural recovery," are not infrequent in the general population [Institute of Medicine. (1990). Broadening the base of treatment for alcohol problems. Washington, DC: National Academy Press]. The goal of this study was to determine if some college students with a history of binge drinking during high school reduced their bingeing without intervention while in college. A second goal was to identify individual characteristics that differentiate between current and reduced bingers. Ninety-one college students with a history of bingeing in high school and no prior drug treatment completed questionnaires about prior and current drinking. Results revealed that 22% of the students with a history of adolescent bingeing had reduced their alcohol consumption while still in college and without treatment. Key factors that differentiated between groups included marital status, church attendance, and outcome and efficacy expectancies. PMID- 12726782 TI - Exercise interventions for smokers with a history of alcoholism: exercise adherence rates and effect of depression on adherence. AB - This study examined the adherence rates and the effect of depression on adherence in two studies conducted among smokers with a past history of alcoholism. In both studies, subjects participated in a 12-session group-based exercise intervention for smoking cessation. The target quit date (TQD) was Session 8. Participants in Study 1 were 73 smokers (43% female). Exercise instructions began at Session 8 and continued through Session 12. Mean frequency and number of minutes of exercise decreased during the 4 weeks of exercise treatment (P<.001). Study 2, conducted with 18 smokers (50% female), examined the feasibility of commencing exercise at Session 1, well before the TQD. The mean number of minutes exercised increased from Sessions 1 to 12 (P=.013). In both studies, average session attendance was high (82%). Combining subjects from both studies, depressed smokers at baseline reported greater mean frequency of exercise per week than nondepressed smokers (P=.05). The results suggest that depressed smokers can be engaged in an exercise program. Further research is needed to determine if commencing exercise early during treatment, prior to the TQD, improves adherence. PMID- 12726783 TI - Stress and the urge to drink. AB - OBJECTIVE: Understanding why people drink alcohol is important for the health and safety of individuals and the public. The aim of this study was to examine from a cognitive point of view the hypothesized link between drinking and stress. METHODS: 25 scenarios were constructed by combining two items, either two life change events or a social situation and an emotional state. In the initial three experiments, 159 male and 43 female alcoholics and 157 male and 93 female nonalcoholics in France judged the degree to which these scenarios were stressful and subsequently the degree to which they stimulated an urge to drink. In the final experiment, 126 of the male alcoholics were studied at the beginning and end of an inpatient alcohol rehabilitation program. RESULTS: The alcoholics and nonalcoholics, regardless of gender, assigned similar stress values to the scenarios and used the same cognitive rules for combining the stress associated with two items (disjunctive rules for two life-change events and additive ones for a personal emotion combined with a social situation). They differed, however, in how they judged the urge to drink. The nonalcoholics reported little stimulus to drink from any combination of items, whereas the alcoholics not only perceived the individual items as stimulating an urge to drink but also used the same cognitive rule in judging the combined urge to drink of two items as they used in judging the combined stress. After completing rehabilitation, the alcoholics judged the combinations of life-change events as stimulating less stress and less urge to drink. Nevertheless, they continued to use a disjunctive combination rule. CONCLUSIONS: Stress and drinking are linked at a fundamental cognitive level among alcoholics, though not among nonalcoholics. Alcoholics should be helped to recognize this link, to reduce their feelings of stress, and to find outlets other than drink. PMID- 12726784 TI - A longitudinal exploration of alcohol use and problems comparing managerial and nonmanagerial men and women. AB - The purpose of the present study was to compare the job attitude and drinking context correlates of alcohol beliefs, consumption, and problems between managerial and nonmanagerial women and men. Using longitudinal, self-report data from 1244 workers in a large manufacturing organization, we found that managerial women reported significantly higher levels of alcohol problems on a number of measures on both Times 1 and 2 surveys. Using partial correlations and controlling for Time 1 levels of the alcohol-related dependent variables, we found that few work attitudes predicted the outcomes of escape drinking reasons, alcohol consumption, and alcohol problems. However, those correlations that were significant reflected a differential pattern for managerial women as compared to managerial men and nonmanagerial women and men. Due to the small subsample size of managerial women, we regard these findings as suggestive only. We discuss the findings in terms of the stress-reduction hypothesis of alcohol consumption. PMID- 12726785 TI - Adolescent alcohol use and suicidal ideation: a nonrecursive model. AB - Prior research has found that adolescent alcohol use is correlated with suicide ideation and behaviors. The causal nature of this relationship, however, has not been established. It could result from a significant causal effect in either direction, both directions, or joint influence from some third factor. These possibilities were addressed using data from a two-wave (24-month) panel survey of junior and senior high school students aged 12 or older at Wave 1. A total of 615 students (301 males, 314 females) completed both waves. Alcohol Problems were related to suicide ideation and behavior cross-sectionally within Waves 1 and 2. Separate Full-Information Maximum Likelihood (FIML) models were estimated for males and females using the same set of identifying restrictions. Results suggested that suicidality leads to increased alcohol-related problems for females, while alcohol-related problems are predictive of suicidality among males. Sensitivity and spuriousness tests did not appreciably alter this conclusion. PMID- 12726786 TI - Predictive value of aspects of the Transtheoretical Model on smoking cessation in a community-based, large-group cognitive behavioral program. AB - OBJECTIVE: To evaluate the predictive value of aspects of the Transtheoretical Model (TTM) of behavior change as applied to smoking cessation in a large-group, community-based cognitive-behavioral intervention. RESEARCH APPROACH: Cognitive behavioral intervention followed by 3-month assessment of smoking status. SETTING: Regional Outpatient Cancer Centre. STUDY PARTICIPANTS: A total of 2069 participants in smoking cessation clinics held between 1992 and 1999. INTERVENTION: Eight 90-min sessions over 4 months utilizing education, self monitoring, a group quit date, and behaviour modification techniques. MAIN OUTCOME MEASURES: Cessation rates at 3 months postquit date. Differences between successful and unsuccessful participants on the baseline TTM variables of: stages of change, processes of change, decisional balance and situational temptations, as well as of precessation demographic, smoking history, and smoking behavior variables. RESULTS: Nonsmokers at 3 months endorsed using more of only one of the processes of change (Reinforcement Management) more than smokers prior to starting the program. They also endorsed more Cons of Smoking and had a more negative Decisional Balance score. When the variables of tobacco tolerance on the Fagerstrom Test for Nicotine Dependence (FTND), marital status, association with the Cancer Centre, and amount of vigorous exercise were first entered in a logistic regression model, Reinforcement Management and Cons of Smoking continued to be predictive of smoking cessation success, but again none of the other TTM variables added explanatory power. CONCLUSIONS: TTM variables measured prior to program attendance added little predictive value for cessation outcome beyond that explained by demographic and smoking history variables. Future studies may benefit from reassessing the TTM variables at the quit date and the 3-month assessment of smoking status to evaluate how the program impacted these variables. PMID- 12726787 TI - A classification of substance-dependent men on temperament and severity variables. AB - This study examined the validity of classifying substance abusers based on temperament and dependence severity, and expanded the scope of typology differences to proximal determinants of use (e.g., expectancies, motives). Patients were interviewed about substance use, depression, and family history of alcohol and drug abuse. Self-report instruments measuring temperament, expectancies, and motives were completed. Participants were 147 male veterans admitted to inpatient substance abuse treatment at a U.S. Department of Veterans Affairs medical center. Cluster analysis identified four types of users with two high substance problem severity and two low substance problem severity groups. Two, high problem severity, early onset groups differed only on the cluster variable of negative affectivity (NA), but showed differences on antisocial personality characteristics, hypochondriasis, and coping motives for alcohol. The two low problem severity groups were distinguished by age of onset and positive affectivity (PA). The late onset, low PA group had a higher incidence of depression, a greater tendency to use substances in solitary contexts, and lower enhancement motives for alcohol compared to the early onset, high PA cluster. The four-cluster solution yielded more distinctions on external criteria than the two cluster solution. Such temperament variation within both high and low severity substance abusers may be important for treatment planning. PMID- 12726788 TI - The association of alcohol and family problems in a remote indigenous Australian community. AB - While a large proportion of Aboriginal Australians do not consume alcohol, those who do frequently show severe alcohol problems. In European-derived samples, heavy alcohol use is associated with relationship distress, conflict, and violence. Because Aboriginal groups commonly have very different family structures, values, and obligations to European-derived families, the association of family and alcohol problems in Aboriginal Australians may be different from European-derived families. This self-report study is the first known published empirical study of family and alcohol problems among Aboriginal Australians. It involved 99 people from a remote community in the far North of Australia. The aims were to explore the association of family conflict, family cohesion, family independence, alcohol problems, and alcohol-related expectancies. Compared to those without alcohol problems, people with alcohol problems reported more family conflict and women with alcohol problems reported high family independence. Expectancies of negative affect change mediated the association of alcohol problems and family conflict. Family cohesion was unrelated to alcohol or family problems. Implications for detection of and interventions for alcohol and family problems are discussed. PMID- 12726789 TI - Adolescent nicotine dependence and smoking cessation outcomes. AB - The purpose of the present study was to examine adolescent nicotine dependence and its impact on smoking cessation outcomes with two treatments of varying intensity: a brief, self-help intervention and an intensive, multisession, school based cessation curriculum called Not On Tobacco (N-O-T). A majority (80%) of adolescent smokers in this study were moderately to highly nicotine-dependent, using the Fagerstrom Tolerance Questionnaire. Further, nicotine dependence was positively correlated with duration of smoking and number of cigarettes smoked daily (P<.05). Data showed that the more cigarettes teens smoked daily and the longer they had smoked, the more dependent they were. Some teens (20%), however, had low nicotine dependence despite years of smoking and high smoking rates. Results showed that the relationship between nicotine dependence and cessation outcomes varied by treatment intensity. The brief intervention was successful with only low-dependent smokers, whereas the intensive, multisession, N-O-T intervention was effective with smokers possessing a range of nicotine dependence, including high-dependent smokers. PMID- 12726790 TI - Self-perceived excessive alcohol consumption among employed women: association with health and psychosocial factors. PMID- 12726792 TI - Validity of self-report of illicit drug use in young hypertensive urban African American males. AB - Inaccurate self-reporting is the primary threat to the validity and utility of self-report in the research on illicit drug use. The purpose of this study was to estimate the sensitivity and specificity of self-report of drug use, using urine toxicity screening as a criterion variable, and to explore the individual characteristics associated with false reporting in a sample of urban African American males. Baseline data from urine tests and a questionnaire about self report of illicit drug use were obtained from 290 hypertensive African American males enrolled in an ongoing hypertension clinical trial. Sensitivity and specificity of self-report were estimated, and multivariate logistic regression analysis was conducted to examine factors associated with true or false reporting of drug use. RESULT: The sensitivity and specificity of self-report were 0.87+/ 0.019 and 0.63+/-0.02, respectively. Members of the underreporting group were more likely to be older, more educated, employed, living with family, and having experience with jail. Jail experience (OR, 2.0) and living with friends or family (OR, 1.7) were the two strongest predictors of false reporting. This study further confirms that social desirability and fear of the consequences of drug use are major contributing factors in underreporting of drug use. PMID- 12726791 TI - Reduction or cessation of injecting risk behaviours? Treatment outcomes at 1-year follow-up. AB - This paper investigates changes in injecting and shared use of needles and syringes among 732 drug misusers recruited to residential and community treatment programmes as part of the National Treatment Outcome Research Study (NTORS). Specifically, it investigates whether reductions in these behaviours were due to a move away from injecting among drug users or whether they could be accounted for by drug users becoming abstinent. Injecting and sharing of injecting equipment were substantially reduced after treatment. Outcomes were due to reduced sharing among injectors, reduced injecting among continuing users and to users becoming abstinent. Clients from residential programmes were more likely to be abstinent at follow-up: methadone clients were more likely to be injecting but not sharing. Abstinence and intermediate risk reduction outcomes were achieved by many drug injectors from both treatment settings. Both outcomes confer benefits, though the benefits of abstinence are greater. Users who were injectors at intake but who had stopped injecting at follow-up consistently achieved superior outcomes across a range of problem behaviours. PMID- 12726793 TI - Parental awareness of adolescent substance use. AB - Parental awareness of adolescent substance use was investigated in a high school sample of 985 adolescents and their parents. Only 39% of parents were aware their adolescent used tobacco, only 34% were aware of alcohol use, and only 11% were aware of illicit drug use. There were no variables that differentiated aware from unaware parents for all substances. Greater parental awareness of alcohol and tobacco use occurred with older adolescents. High adolescent ratings of family communication combined with low parental ratings of family communication were also associated with greater parental awareness of alcohol and tobacco use. Better school grades predicted greater awareness of alcohol and illicit drug use. Single parents and blended families were more aware of tobacco and illicit drug use. PMID- 12726794 TI - Interest in self-help materials among a general population sample of smokers. AB - Previous studies have shown that self-help materials can be effective in helping people quit smoking. However, it is not known what proportion of smokers in the general population are interested in this method of cessation. A representative sample of 267 daily smokers participated in a random digit dialing telephone survey. Respondents were asked about their level of interest in self-help materials that would help them quit smoking. Potential predictors of level of interest were examined first through bivariate analyses, followed by a multinomial logistic regression. Of daily smokers, 27% were very interested in receiving a self-help booklet, 28% were somewhat interested, and 45% were not at all interested. Interest in self-help materials was positively related to intent to quit in the next 6 months and to low family income. PMID- 12726795 TI - Irritability following abstinence from cocaine predicts euphoric effects of cocaine administration. AB - This study evaluated the association between negative affective symptoms during initial abstinence and euphorigenic response to experimentally administered cocaine. Cocaine-dependent individuals achieved 5 days of abstinence in a hospital setting. Forty milligrams of cocaine was given intravenously on the fifth day of abstinence, and participants were asked to rate the subjective effects produced by the drug. The associations between irritability, self reported depression, and the subjective "high" produced by cocaine were evaluated. Increased levels of irritability and depression both correlated positively and statistically significantly with heightened response to experimentally administered cocaine as indexed by self-reported subjective "high." The positive association between irritability and subjective "high" remained after controlling for self-reported depressive symptoms. The opponent process model predicts that increased levels of negative affect should be associated with diminished euphoric response to cocaine; however, the opposite was observed. If these findings are replicated in a larger sample, then it may be necessary to reconsider the applicability of the opponent process model to cocaine addiction in humans. PMID- 12726797 TI - The relationship between structure and function: why does reshaping the left ventricle surgically not always result in functional improvement? AB - Surgical strategies recently introduced to improve ventricular function have been based on the concepts of reduction of ventricular diameter, synchronization of myocardial activity, passive support of diastolic ventricular shape, and active support of systolic ventricular constriction. They have depended on several established theoretical assumptions, not all of which are totally valid. Clinical results have proved markedly variable. This is especially true for procedures designed to reduce the radius of the left ventricle. Some have reported up to 80% mortality, whereas others achieve results comparable with those for heart transplantation. Because of this, the method runs the risk to be rejected, or else, its more widespread application will be postponed until essential details concerning the basic concepts have been elucidated. It is these details which we discuss in this review. PMID- 12726798 TI - Shape of the left ventricle and its computer modelling. AB - A simple computer program was made to draw different left ventricle shapes in order to support the theory of elongation and to get a visual presentation of the shape of the left ventricle. Experimental data, obtained from echocardiography and Simpson's rule, were used for this program. The results yielded different shapes under different physiological circumstances, indicating the sensitivity of the method. It was concluded that these figures (shapes) support the use of elongation as a shape index. PMID- 12726799 TI - Topical cardiac cooling--computer simulation of myocardial temperature changes. AB - Topical cardiac cooling (TC) is often used in cardiac surgery. We used a computer simulation to study temperature changes in the heart, especially in the right ventricular wall. A three-dimensional computer heart model, derived from Visible Human Data set, National Library of Medicine was used. The model is made from cubes, with spatial resolution of 1mm. Explicit Finite Different method and temperature diffusion equation were used to calculate new temperatures. Three different simulations were performed and simulated temperatures were drawn on a cross-section of heart model in different colors. The results show that areas not immersed into TC solution are less protected against ischemia. It is important not to rely solely on topical cooling but use appropriate method of myocardial protection technique. PMID- 12726801 TI - Changes of the beat amplitude power after partial left ventriculectomy and coronary artery bypass grafting. AB - Partial left ventriculectomy (PLV) was originally introduced as a new surgical approach by patients with an end stage of cardiac disease. Coronary artery bypass grafting (CABG) is a standard procedures used in cardiac surgery. Multichannel ECG (MECG) measurements and body surface mapping (BSM) were used to analyse the normalised beat amplitude power maps (BAM) that reflect an overall cardiac activity. The resulting BAMs show that the amplitude of cardiac signals decrease for approximately 30% after the PLV and stay in the same level during the postoperative monitoring interval while after CABG no significant changes in BAM have been observed. In addition the electrodes from the body surface area above the left ventricle, where surgery was performed, show significant changes in beat amplitudes. PMID- 12726800 TI - Computer analysis of multichannel ECG. AB - Multichannel electrocardiography (MECG) is an extension of the conventional electrocardiography that is aimed at refining the non-invasive characterisation of cardiac activity. Body surface mapping is a graphical presentation of cardiac activity as measured from the body surface. Body surface maps can show the distribution of the potential at a selected moment in time or over a specified time interval. A new family of maps, based on the characteristics derived from the complete analysed beat, is described. Some new computer supported methods, which are able to calculate automatically different temporal maps, are proposed. MECG measurements can be seen in this context as a powerful research and clinical tool for improving the resolution of cardiac measurements. PMID- 12726802 TI - Body surface mapping of cardiac activity after partial left ventriculectomy. AB - MECG measurements were performed using 35 electrodes in 10 patients operated with partial left ventriculectomy (PLV). Body surface ECG signals were recorded and five measurements were done: prior to PLV, second, third, fourth and fifth postoperative day. This work was concentrated on the following mapping methods: average isopotential ST segment maps (STM), QRS interval isointegral maps (QRM) and isochronal activation maps (IAM). STMs of the patients show a great positive area (elevation) over the anterior aspect of the heart and a great negative area (depression) over the lateral and posterior aspect of the heart before the operation. After the operation, the ST elevation over the anterior, lateral and posterior aspect of the heart was reduced. A substantial positive value over the excised area of the heart was present also on the end of the postoperative monitoring interval. Minimal and maximal values of the QRMs were smaller and also show some kind of normalisation. The area of the left ventricle, where PLV was performed, was carefully analysed for any changes of activation time for different heart regions. IAMs indicate that the start of the first activation was quite stable and in accordance with the position of the QRMs minimum. PMID- 12726803 TI - The effect of nontransmural necroses on epicardial potential maps during paced activation: a simulation study. AB - A number of studies have indicated that epicardial potentials provide detailed spatiotemporal information about the spread of electrical activation within the ventricular wall. Here, we used a computer model to simulate activation sequences and corresponding epicardial potential maps in the ventricles damaged by localized necroses. Our findings agreed with those of experimental studies performed for epicardial pacing locus in a complete transient loss of one of the positive areas when the necrosis was located subepicardially, and in a transient gap in the expanding positive areas when the necrosis was located intramurally and subendocardially. This study--by systematically comparing simulated epicardial potential maps with those recorded on the exposed canine hearts- constitutes an important step in validation of our model. PMID- 12726804 TI - Breathing rates and heart rate spectrograms regarding body position in normal subjects. AB - The right lateral body position has been proposed as an effective vagal enhancer. However, the possibility of breathing affecting heart rate power spectra in different body positions has not been assessed. The level of vagal modulation in various body positions in normal subjects was estimated by calculating heart rate power spectra. The results suggest that the levels of vagal modulation do not necessarily reflect a change due to assuming different body position, but might be the consequence of changed breathing patterns. Before adopting the right lateral body position as vagal enhancing, the contribution of varying breathing pattern should be eliminated. PMID- 12726805 TI - Beat-to-beat QT interval variability before and after cardiac surgery. AB - Non-uniform recovery of excitability may be essential in triggering malignant ventricular tachycardia after cardiac surgery. Thirty-five channels ECG was recorded for 6 min in 27 patients before and after heart surgery and in 20 control subjects. Off-line analysis was performed. RR interval duration, RR SD, QT SD and power spectra of RR variability were computed from 256 s stable RR and QT interval series. When compared to controls, patients had decreased RR SD and increased QT SD before surgery (p<0.002 and p<0.0005, respectively); RR SD further decreased and QT SD increased after the surgery (p<0.0001 and p<0.0002, respectively). Increase of QT variability and decrease of RR variability after cardiac surgery may reflect disrupted electrophysiological stability of the myocardium and thus electrophysiological substrate for triggering malignant arrhythmia. PMID- 12726807 TI - Virtual coronary cineangiography. AB - The mastering of myocardial infarction diagnosis is traditionally composed of laborious trial- and error-based examination of canonical coronary cineangiographies. In the following article we suggest a system that enables the instructor to generate student-specific cases, thus allowing teaching not only the basic feature searching and stenosis evaluation processes, but also the importance of the correct acquisition viewpoint. With the proposal of the development of the Digital Cardiologist intelligent agent we also envisage the possibility of the student's self-tutoring. PMID- 12726806 TI - Confidential storage and transmission of medical image data. AB - We discuss computationally efficient techniques for confidential storage and transmission of medical image data. Two types of partial encryption techniques based on AES are proposed. The first encrypts a subset of bitplanes of plain image data whereas the second encrypts parts of the JPEG2000 bitstream. We find that encrypting between 20% and 50% of the visual data is sufficient to provide high confidentiality. PMID- 12726808 TI - Childhood-onset neuropsychiatric disorders among adult patients in a Swedish special hospital. PMID- 12726809 TI - Courts, doctors, and insanity defences in 18th and early 19th century Scotland. PMID- 12726810 TI - Patients' perceptions of coercion on admission to forensic psychiatric hospital: a comparison study. PMID- 12726811 TI - Homicides of patients in hospitals and nursing homes: a comparative analysis of case series. PMID- 12726813 TI - Testamentary capacity and suicide: an overview of legal and psychiatric issues. PMID- 12726812 TI - Declining trends in U.S. parricides, 1976-1998: testing the Freudian assumptions. PMID- 12726814 TI - The characteristics of criminal and noncriminal mentally disordered patients. PMID- 12726815 TI - Harassment and intimidation of forensic psychiatrists: an update. PMID- 12726816 TI - Leptin in the CNS: much more than a satiety signal. AB - The discovery of the obese gene product, leptin has generated enormous interest in how the periphery signals the status of nutritional stores to specific hypothalamic nuclei involved in regulating feeding and energy balance. However it is emerging that leptin, in addition to its role as a circulating satiety factor, is a multi-faceted hormone that plays a key role in a variety of CNS functions. In this review, we summarise recent progress in leptin biology, with particular focus on its diversity of actions within the CNS, ranging from satiety signal, to regulator of bone formation and inhibitor of neuronal excitability. PMID- 12726817 TI - Insulin inhibits rat hippocampal neurones via activation of ATP-sensitive K+ and large conductance Ca2+-activated K+ channels. AB - In this study, we have used a combination of immunocytochemical and Ca(2+) imaging techniques to determine the functional localisation of insulin receptors as well as the potential role for insulin in modulating hippocampal synaptic activity. Comparison of insulin receptor and MAP2 labelling demonstrated extensive insulin receptor immunoreactivity on the soma and dendrites of cultured hippocampal neurones. Dual labelling with synapsin 1 also showed punctate insulin receptor labelling associated with synapses. In functional studies, insulin inhibited spontaneous Ca(2+) oscillations evoked in cultured hippocampal neurones following Mg(2+) removal. This action of insulin was mimicked by the ATP sensitive K(+) (K(ATP)) channel opener diazoxide or the large conductance Ca(2+) activated K(+) (BK) channel activator NS-1619. Furthermore, application of the K(ATP) channel blocker glybenclamide or the BK channel inhibitors iberiotoxin or charybdotoxin attenuated the actions of insulin, whereas prior incubation with a combination of glybenclamide and iberiotoxin completely blocked insulin action. The ability of insulin to modulate the Ca(2+) oscillations was reduced by the inhibitors of MAPK activation PD 98059 and U0126, but not by the PI 3-kinase inhibitors LY 294002 or wortmannin, indicating that a MAPK-driven process underlies insulin action. In conclusion, insulin inhibits spontaneous Ca(2+) oscillations via a process involving MAPK-driven activation of BK and K(ATP) channels. This process may be a useful therapeutic target for the treatment of epilepsy and certain neurodegenerative diseases. PMID- 12726819 TI - Mutation in a protein kinase C phosphorylation site of the 5-HT1A receptor preferentially attenuates Ca2+ responses to partial as opposed to higher-efficacy 5-HT1A agonists. AB - The Thr(149)Ala mutation in a putative protein kinase C phosphorylation site of the 5-HT(1A) receptor's second intracellular loop has been shown to affect the closing of Ca(2+) channels and Ca(2+) mobilisation without interfering with the inhibitory cAMP pathway (Mol Pharmacol 52 (1997) 164). Here, the Ca(2+) responses for a series of 5-HT(1A) agonists were compared between the wild-type (wt) and mutant Thr(149)Ala 5-HT(1A) receptor as part of a fusion protein containing a G(alpha)(15) protein. Neither the mutation nor the fusion process modified the [(3)H]WAY 100635-based ligand binding profile of the fusion proteins as compared to the wt 5-HT(1A) receptor protein. Whereas at the wt 5-HT(1A) receptor, 5-HT induced a Ca(2+) response in CHO-K1 cells via endogenous G(i/o) proteins, the Ca(2+) response to 5-HT at the mutant Thr(149)Ala 5-HT(1A) receptor was fully dependent on either the co-expression or the fusion to a recombinant G(alpha)(15) protein. Buspirone, flesinoxan and 8-OH-DPAT produced a graded partial response (26 to 62%) at the wt 5-HT(1A):G(alpha)(15) fusion protein; F 13640, 5-CT and F 14679 behaved as higher-efficacy agonists with maximal Ca(2+) responses similar to 5-HT. The maximal Ca(2+) responses at the mutant Thr(149)Ala 5 HT(1A):G(alpha)(15) fusion protein were significantly attenuated for flesinoxan and 8-OH-DPAT (-45 and -36%, respectively); the response to the other 5-HT agonists was not significantly affected. A similar effect was observed upon treatment with phorbol 12-myristate 13-acetate at the Thr(149)Ala 5 HT(1A):G(alpha)(15) fusion protein. In conclusion, the amplitude of the Ca(2+) responses induced by partial, but not that to fuller 5-HT(1A) receptor agonists, is affected by the Thr(149)Ala mutation of the 5-HT(1A):G(alpha)(15) fusion protein. PMID- 12726818 TI - The effects of general anaesthetics on carbachol-evoked gamma oscillations in the rat hippocampus in vitro. AB - The effects of general anaesthetics and temperature on carbachol-evoked gamma oscillations in the rat hippocampal brain slice preparation were investigated. The frequency of the oscillations was found to be dependent on temperature in the range 32-25 degrees C, with a linear reduction in frequency from 40-17 Hz over this temperature range. The volatile anaesthetics isoflurane and halothane, and the intravenous anaesthetics thiopental, propofol and R(+)-etomidate caused a reduction in the frequency of the oscillations, in a concentration-dependent manner, over a range of clinically relevant concentrations. On the other hand, the intravenous agent ketamine and the "inactive" S(-)-isomer of etomidate had no significant effect on the oscillation frequency. The oscillations were markedly asymmetric over one cycle with a relatively rapid "rising" phase followed by a slower "decaying" phase. The decrease in oscillation frequency was due to an increase in the time-course of the "decaying phase" of the oscillation with little effect on the "rising" phase, consistent with the idea that carbachol evoked gamma oscillations are trains of GABAergic inhibitory postsynaptic potentials and that the anaesthetics are acting postsynaptically at the GABA(A) receptor. PMID- 12726820 TI - Alpha2-adrenergic drug effects on brain monoamines, locomotion, and body temperature are largely abolished in mice lacking the alpha2A-adrenoceptor subtype. AB - alpha(2)-ARs regulate brain monoaminergic function by inhibiting neuronal firing and release of monoamine neurotransmitters, noradrenaline (NA), serotonin (5-HT) and dopamine (DA). Both alpha(2A)- and alpha(2C)-AR inhibit monoamine release in vitro in brain slices, but the in vivo roles of individual alpha(2)-AR subtypes in modulating monoamine metabolism have not been characterised. Metabolism of brain monoamine neurotransmitters, locomotor activity and body temperature were investigated in mice with targeted inactivation of the gene encoding alpha(2A)-AR (alpha(2A)-knockout, alpha(2A)-KO) and wild-type (WT) mice after treatment with the alpha(2)-AR agonist dexmedetomidine and the antagonist atipamezole. Dexmedetomidine caused profound hypothermia (up to 14.7 degrees C mean reduction in rectal temperature) and locomotor inhibition in WT mice, and inhibited the turnover of NA, 5-HT and DA, but increased NA turnover in alpha(2A)-KO mice. alpha(2)-AR agonist-induced hypothermia and locomotor inhibition were attenuated, but not totally abolished, in alpha(2A)-KO mice. These results suggest that alpha(2A)-ARs are principally responsible for the alpha(2)-AR mediated inhibition of brain monoamine metabolism, but other alpha(2)-ARs, possibly alpha(2C)-ARs, are also involved, especially in the striatum. However, secondary effects of the physiological alterations caused by drug administration, especially hypothermia, may have contributed to the observed neurochemical changes in WT mice. PMID- 12726821 TI - Effect of different 5-HT1A receptor antagonists in combination with paroxetine on expression of the immediate-early gene Arc in rat brain. AB - Selective 5-HT(1A) receptor antagonists enhance the effect of selective serotonin reuptake inhibitors (SSRIs) on presynaptic 5-HT function, and have potential as antidepressant augmentation therapies. The present study tested the effect of different selective 5-HT(1A) receptor antagonists (WAY 100635, NAD-299, p-MPPI and LY 426965) in combination with a SSRI (paroxetine), on postsynaptic 5-HT function measured by increased expression of the immediate early gene, Arc. Paroxetine (5 mg/kg s.c.) combined with WAY 100635 (0.3 mg/kg s.c.) increased Arc mRNA in frontal, parietal and piriform cortices, and caudate putamen. Paroxetine (5 mg/kg s.c.) plus NAD-299 (1 or 5 mg/kg s.c.) had a similar effect. None of these drugs increased Arc mRNA when administered alone. Paroxetine (5 mg/kg s.c.) plus p-MPPI (8.5 mg/kg s.c.) also increased Arc mRNA but p-MPPI itself elevated Arc mRNA in many regions. Whilst LY 426965 (3 or 10 mg/kg s.c.) had no effect alone, when combined with paroxetine (5 mg/kg s.c.), the drug increased Arc mRNA in caudate putamen but not cortical regions.In conclusion, this study demonstrates that four 5-HT(1A) receptor antagonists augment the effect of an SSRI on Arc mRNA expression, which is suggestive of increased postsynaptic 5-HT function. However, the data reveal certain differences in the 5-HT(1A) receptor antagonists not recognised in models of presynaptic 5-HT function. PMID- 12726822 TI - Bi-phasic change in BDNF gene expression following antidepressant drug treatment. AB - The gene for brain derived neurotrophic factor (BDNF) has recently received attention in relation to the therapeutic action of antidepressant treatment. This study aimed to clarify the influence of post drug interval on the effect of acute and repeated treatment with antidepressant drugs on BDNF gene expression in the rat brain. It was found that repeated administration of either the monoamine oxidase inhibitor tranylcypromine (TCP) or 5-hydroxytryptamine (5-HT) re-uptake inhibitors (fluoxetine, paroxetine and sertraline), evoke a bi-phasic and time dependent effect on BDNF gene expression in the rat hippocampus (especially dentate gyrus). A down-regulation of the BDNF gene was detected at 4 h (TCP and fluoxetine) and an up-regulation at 24 h (TCP, paroxetine, fluoxetine, sertraline) after the last of twice daily injections for 14 days. After a single injection the down-regulation was detected at 4 h (TCP, fluoxetine, paroxetine and sertraline) but BDNF mRNA levels were not altered at 24 h post drug (TCP, fluoxetine and paroxetine). Administration of inhibitors of noradrenaline re uptake (desipramine and maprotiline) or the atypical antidepressant mianserin had no effect on BDNF mRNA levels at either single (4 h post drug, desipramine) or repeated (24 h post drug, desipramine, maprotiline, mianserin) treatment. The gene expression for NT-3, which is distributed in a high density in the dentate gyrus, was not affected by single or repeated injections of antidepressant drugs (TCP, fluoxetine, paroxetine, sertraline, desipramine, maprotiline or mianserin) at 4 or 24 h post drug. In conclusion, these data show that the effect of antidepressant drugs on BDNF gene expression may be more complex and less widespread across treatments than previously thought. Thus, in this study drugs interacting with the central 5-HT system altered BDNF expression but the effect was bi-phasic over the 24 h post drug period. PMID- 12726823 TI - Effects of single and repeated electroconvulsive shock on the social and agonistic behaviour of resident rats. AB - The aim of this study was to determine whether electroconvulsive shock (ECS, an established antidepressant treatment), like acute and chronic antidepressant drug treatments, produces similar differential effects on the behavioural profile of resident rats expressed during social encounters with unfamiliar intruder conspecifics (resident-intruder paradigm). Thirty minute pretreatment with a single ECS suppressed both investigation and aggression directed at intruders concomitant with increased flight behaviour and marked sedation. Behavioural disruption subsided over the following 24 h. In contrast, resident rats subjected to bi-daily ECS treatment expressed elevated aggression at days 7 (four shocks) and 14 (eight shocks). Eight days after the last ECS treatment the behaviour of the resident rats had returned to pretreatment values. Additional studies showed that bi-daily ECS treatment nearly abolished 5-HT(2C) receptor-mediated hypolocomotion induced by acute m-chlorophenylpiperazine (mCPP, 2.5 mg/kg sc) challenge 24 h following 2 ECSs, while 4 ECSs only enhanced 5-HT(2A) receptor mediated head shakes induced by 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI, 2.0 mg/kg sc). These studies demonstrate that repeated ECS treatment increases the aggressive behaviour of resident rats which may be associated with adaptive changes in 5-HT(2C) and 5-HT(2A) receptor-mediated function. It remains to be seen whether adaptive changes in 5-HT(2C) receptor function represent a common mechanism of clinical antidepressant efficacy. PMID- 12726824 TI - Amphetamine withdrawal modulates FosB expression in mesolimbic dopaminergic target nuclei: effects of different schedules of administration. AB - Different patterns of psychostimulant intake can elicit widely varying behavioral and neurochemical consequences. Accordingly, rats were studied during withdrawal from either of two schedules of amphetamine administration, one consisting of 6 days of low-dose (1.5 mg/kg, i.p.) daily intermittent (INT) amphetamine (AMPH) injections, and the other of 6 days of moderately high-dose (1-5 mg/kg, i.p.) escalating (ESC) AMPH injections, for the effects of these treatments on numbers of FosB-positive nuclei and monoamine utilization in dopaminergic target areas. Withdrawal from AMPH pretreatment according to the ESC schedule markedly increased FosB expression in the nucleus accumbens shell and basolateral amygdala. In contrast, withdrawal from INT-AMPH administration did not increase FosB expression in any of the regions examined. Post-mortem neurochemical analyses of these same brain regions did not reveal effects of withdrawal from either INT or ESC administration of AMPH. These results suggest that withdrawal from a moderately high-dose AMPH regimen modifies patterns of gene expression in mesocorticolimbic dopaminergic target nuclei without significantly affecting basal monoamine levels. The strength of these effects in the nucleus accumbens shell and basolateral nucleus of the amygdala are consistent with behavioral and clinical data indicating the importance of these areas in the neuroadaptive changes which characterize addiction and withdrawal states. PMID- 12726826 TI - Effects of the nitric oxide donor, DEA/NO on cortical spreading depression. AB - Cortical spreading depression (CSD) is a transient disruption of local ionic homeostasis that may promote migraine attacks and the progression of stroke lesions. We reported previously that the local inhibition of nitric oxide (NO) synthesis with Nomega-nitro-L-arginine methyl ester (L-NAME) delayed markedly the initiation of the recovery of ionic homeostasis from CSD. Here we describe a novel method for selective, controlled generation of exogenous NO in a functioning brain region. It is based on microdialysis perfusion of the NO donor, 2-(N,N-diethylamino)-diazenolate-2-oxide (DEA/NO). As DEA/NO does not generate NO at alkaline pH, and as the brain has a strong acid-base buffering capacity, DEA/NO was perfused in a medium adjusted at alkaline (but unbuffered) pH. Without DEA/NO, such a microdialysis perfusion medium did not alter CSD. DEA/NO (1, 10 and 100 microM) had little effect on CSD by itself, but it reversed in a concentration-dependent manner the effects of NOS inhibition by 1 mM L-NAME. These data demonstrate that increased formation of endogenous NO associated with CSD is critical for subsequent, rapid recovery of cellular ionic homeostasis. In this case, the molecular targets for NO may be located either on brain cells to suppress mechanisms directly involved in CSD genesis, or on local blood vessels to couple flow to the increased energy demand associated with CSD. PMID- 12726825 TI - Effects of the cocaine- and amphetamine-regulated transcript peptide on the turnover of central dopaminergic neurons. AB - The effects of the cocaine- and amphetamine-regulated transcript (CART) peptide on central dopaminergic (DA) neurons were examined in ovariectomized, estrogen primed Sprague-Dawley rats in both the morning and afternoon. Intracerebroventricular administration of 1 microg, but not lower doses of the CART peptide (55-102), either in the morning or afternoon produced a prolonged increase in the 3,4-dihydroxyphenylacetic acid (DOPAC) level in the median eminence (ME) and a corresponding decrease of serum prolactin (PRL) levels, which resulted from stimulation of tuberoinfundibular dopaminergic neurons. The CART peptide stimulated DOPAC levels in the striatum (ST), nucleus accumbens (NA), hypothalamic paraventricular nucleus (PVN), and periventricular (A14), but had no effect in the medial prefrontal cortex (MPFC) or suprachiasmatic nucleus (SCN). These effects of the CART peptide on stimulation of central DA systems and inhibition of PRL levels are specific because the inactive form of the CART peptide (0.1 and 1 microg) could not induce a similar response. Stimulatory effects of the CART peptide on different central DA systems displayed differential time-response profiles in the NA and ST, ME, and PVN and A14. These findings indicate that the CART peptide may selectively regulate certain central DA neuronal activities. PMID- 12726827 TI - TRPV1 activation and induction of nociceptive response by a non-pungent capsaicin like compound, capsiate. AB - Capsiate is a capsaicin-like ingredient of a non-pungent cultivar of red pepper, CH-19 sweet. To elucidate the mechanisms underlying the non-pungency of capsiate, we investigated whether capsiate activates the cloned capsaicin receptor, TRPV1 (VR1). In patch-clamp experiments, capsiate was found to activate TRPV1 expressed transiently in HEK293 cells with a similar potency as capsaicin. Capsiate induced nociceptive responses in mice when injected subcutaneously into their hindpaws with a similar dose dependency as capsaicin. These data indicate that the non pungent capsiate is an agonist for TRPV1 and could excite peripheral nociceptors. In contrast to this, capsiate did not induce any significant responses when applied to the skin surface, eye or oral cavity of mice, suggesting that capsiate requires direct access to nerve endings to exhibit its effects. Capsiate was proved to have high lipophilicity and to be easily broken down in normal aqueous conditions, leading to less accessibility to nociceptors. Another highly lipophilic capsaicin analogue, olvanil, was similar to capsiate in that it did not produce irritant responses when applied to the skin surface, although it could activate TRPV1. Taken together, high lipophilicity and instability might be critical determinants for pungency and so help in understanding the effects of capsaicin-related compounds. PMID- 12726830 TI - Luminance mechanisms mediate the motion of red-green isoluminant gratings: the role of "temporal chromatic aberration". AB - In this paper we use a dynamic noise-masking paradigm to explore the nature of the mechanisms mediating the motion perception of drifting isoluminant red-green gratings. We compare contrast thresholds for the detection and direction discrimination of drifting gratings (1.5 cpd), over a range of temporal frequencies (0.5-9 Hz) in the presence of variable luminance or chromatic noise. In the first experiment, we used dynamic luminance noise to show that direction thresholds for red-green grating motion are masked by luminance noise over the entire temporal range tested, whereas detection thresholds are unaffected. This result indicates that the motion of nominally isoluminant red-green gratings is mediated by luminance signals. We suggest that stimulus-based luminance artifacts are not responsible for this effect because there is no masking of the detection thresholds. Instead we propose that chromatic motion thresholds for red-green isoluminant gratings are mediated by dynamic luminance artifacts that have an internal, physiological origin. We have termed these "temporal chromatic aberration". In the second experiment, we used dynamic chromatic noise masking to test for a chromatic contribution to red-green grating motion. We were unable to find conclusive evidence for a contribution of chromatic mechanisms to the chromatic grating motion, although a contribution at very high chromatic contrasts cannot be ruled out. Our results add to a growing body of evidence indicating the presence of dynamic, internal luminance artifacts in the motion of chromatic stimuli and we show that these occur even at very low temporal rates. Our results are compatible with our previous work indicating the absence of a chromatic mechanism for first order (quasi-linear) apparent motion [Vision Res. 40 (2000) 1993]. We conclude that previous conclusions based on the motion of chromatic red-green gratings should be reassessed to determine the contribution of dynamic luminance artifacts. PMID- 12726829 TI - Orientation discrimination in visual noise using global and local stimuli. AB - We have investigated orientation discrimination in visual noise using two types of high contrast, broadband stimuli. Discrimination thresholds are better for Local stimuli, in which the orientation signal is spatially limited, than for Global stimuli, in which the orientation signal extends across the entire stimulus. Performance improves with increasing stimulus area, reaching an optimum threshold of about 11% orientation signal. Thresholds were not influenced by brief presentation times or practice. These results, along with results from a simple computational model, suggest that human orientation discrimination for this kind of pattern is mediated by pooling local responses of low-level neural mechanisms and is limited by two stages of intrinsic neural noise. PMID- 12726828 TI - Reexamination of the role of hyperpolarization-activated cation channels in short and long-term plasticity at hippocampal mossy fiber synapses. AB - We tested a proposal that the hyperpolarization-activated cation channel (I(h) channel) is involved in the induction of short- and long-term plasticity at the hippocampal mossy fiber-CA3 synapses. Bath application of a specific I(h) channel blocker ZD 7288, at a concentration at which it blocked I(h) channels, substantially depressed mossy fiber synaptic transmission, and this inhibition was occluded by previous blockade of these channels by CsCl. In addition, ZD 7288 attenuated the amplitude of both AMPA and NMDA receptor-mediated excitatory postsynaptic currents (EPSCs) equally and caused a coincident increase in the failure rate of single-fiber EPSCs and paired-pulse facilitation (PPF). It also blocked long-term potentiation (LTP) induction when applied before high-frequency tetanic stimulation (TS), and reversed LTP when applied afterwards. Continuous application of CsCl, which efficiently blocks I(h) channels, mimicked ZD 7288 in inhibiting LTP. Furthermore, ZD 7288 blocked both forskolin- and Sp-8-CPT-cAMPS mediated enhancements of synaptic transmission. However, it did not affect the frequency facilitation induced by increasing the stimulus frequency from 0.05-1 Hz and the expression of the long-term depression (LTD) induced by low-frequency stimulation (LFS) or DCG-IV. Perforated patch-clamp recordings from granule cells revealed that the voltage for half-maximal activation (V(1/2)) of I(h) was significantly shifted towards the depolarizing direction following forskolin or Sp-8-CPT-cAMPS treatment. This enhanced I(h) current was not due to persistent activation of protein kinase A (PKA), because PKA inhibitor KT5720 did not abolish the difference between the activation curves. Therefore, we conclude that I(h) channels may contribute to the development and regulation of short- and long term plasticity at the mossy fiber-CA3 synapses. PMID- 12726831 TI - Binocular recognition summation in the peripheral visual field: contrast and orientation dependence. AB - Spatial frequency thresholds for recognition were measured for binocular and monocular viewing conditions at two contrast levels (95% and 7%). Measurements were obtained at the fovea and at four different eccentric retinal locations. Each eccentric retinal location was 8 degrees from the fovea, one on the horizontal axis (180 degrees ), and the other three in the superior field on retinal axes of 90 degrees, 45 degrees and 135 degrees. For the superior and horizontal retinal locations, the orientations of the gratings tested were horizontal (180 degrees ) and vertical (90 degrees ). For the retinal points on the oblique axes, the orientations of the gratings were 45 degrees and 135 degrees. Measurements were also obtained at the fovea for all four different grating orientations at both levels of contrast. Recognition threshold was defined as the highest spatial frequency at which luminance gratings were perceived vertically. At the fovea, binocular summation ratios (binocular spatial frequency/monocular spatial frequency) showed no significant differences for gratings of either contrast levels or for any orientation (p>0.05). In the superior periphery, significantly higher summation ratios were shown by low contrast vertical gratings (p<0.05), and in the horizontal periphery by low contrast horizontal gratings (p<0.05). On the oblique axis, low contrast gratings that were parallel to the oblique meridian showed higher summation ratios compared to those at right angles to it. High contrast gratings, at least at 8 degrees eccentricity, did not show this effect. Data suggest that meridional organisation of the retina (e.g. vertical gratings seen maximally in the superior field) occurs for resolution and that it is evidenced closer to the fovea than previously shown. PMID- 12726832 TI - Discrimination of single bars by the honeybee (Apis mellifera). AB - The bees learn to come for a reward to a very simple pattern, a black bar in a fixed position on a white background, in a Y-choice apparatus, with the targets presented in the vertical plane at a fixed range. They were trained on a number of different arrangements of a single bar on one or both targets. The trained bees were then given appropriate tests to discover what cues they had learned. A cue is an essential parameter that is recognized, not the whole pattern. At the choice point they learn exactly which way to look for consistent cues. After training on a single broad bar versus a blank target, they respond in tests to any area of black where they expect to see it, and are less able to detect it the more it has been displaced from the training position. They are more sensitive to vertical than to horizontal displacement of the bar. The cue is anything black of the right size. They do not recognize the shape or orientation of the bar. When trained to discriminate between two bars at right angles to each other, centred on the reward hole, the cue is the edge orientation at the expected places on the targets, and the bees are less able to discriminate the orientation cues the more they are displaced. When trained on a pair of broad black bars in different positions, the cues are the vertical positions of the centres. Division of the bar into squares, or making the edges stepped, removes the orientation cue but not the position cue. Addition of a large black spot or a checkerboard background to the original bar prevents discrimination, as if the spatial reference frame is disturbed. In training, or testing trained bees, parallax does not assist the discrimination of orientation. PMID- 12726833 TI - Wavelength dependence of the optomotor response in zebrafish (Danio rerio). AB - The action spectrum of motion detection in zebrafish (Danio rerio) was measured using the optomotor response in the light adapted state. The function has a single maximum at 550-600 nm, and is similar to the spectral sensitivity function of the L-cone type in the mid and long wavelength range. At shorter wavelengths the values of three of the five fish tested are lower. As in goldfish [Vis. Res. 36 (1996) 4025], the result indicates a dominance of the L-cone type with an inhibitory influence of M- or S-cones. Experiments with a red/green striped cylinder showed that the optomotor response was at minimum whenever the L-cone type was not modulated by the moving pattern. This demonstrates that motion vision in zebrafish is "color blind", using mainly one of the four cone types probably involved in color vision. PMID- 12726834 TI - Synaptic energy efficiency in retinal processing. AB - Recent work suggests that the visual system may represent early visual information in an energy efficient manner [Nature 381 (1996); Nature, 381 (1996) 607; Neural Comput. 3 (2001) 799; Curr. Opin. Neurobiol. 11 (2001) 475]. This paper applies the idea of energy efficient representations to understand retinal processing, and provides evidence that centre surround processing observed is efficient in terms of minimizing synaptic activity. In particular, it is shown that receptive fields at different retinal eccentricities and at different levels of noise, can be understood in terms of maximizing the transmission of visual information given a constraint on total synaptic strengths and hence energy consumption. PMID- 12726835 TI - Adaptation of response transients in fly motion vision. I: Experiments. AB - Two types of transient responses have been investigated in fly motion-sensitive neurons in the past: the impulse and the step response. In response to a brief motion pulse, cells show a sudden rise in activity followed by an exponential decay ('impulse response'). In response to the onset of a constant velocity stimulus, cells exhibit transient oscillations before settling to a steady-state value ('step response'). Since the impulse response has been shown to shorten when tested after presentation of an adapting motion stimulus, we investigated whether adaptation also occurs during the step response. We tested this hypothesis by recording extracellularly the response of the H1-cell in the lobula plate of the blowfly Calliphora vicina to gratings of varying pattern contrasts and drift velocity. We found that the transient oscillations of the step response strongly depend on the pattern contrast: at low contrasts, oscillations lasted for several seconds, whereas at high contrasts, they settled within fractions of a second. This suggests that motion adaptation occurs during the initial period of the stimulus presentation and is dependent on the contrast of the motion stimulus. Using identical stimulus parameters (contrast and temporal frequency) for the adapting stimulus and testing the impulse response afterwards, we found that the impulse response and the transient period in the step response shortened in a similar way. We then analyzed the dynamic of the transients oscillations produced by ongoing motion of a square wave pattern in the anti-preferred direction (null direction) of H1. As observed for preferred direction motion, we found that the duration and amplitude of those transients shortened as the contrast and the velocity of the pattern increased, and that the oscillations disappeared when a blank screen instead of a pattern was presented before the onset of motion. Under both stimulus conditions, i.e. grating and blank screen before motion onset, the steady-state response level showed the same dependence on the contrast and temporal frequency of the pattern. When we analyzed the responses of the cell to pattern of various sizes and contrasts moving in the preferred direction of the cell, we found that increments in the size affected the overall amplitude of both the transient oscillations and the steady-state response level, whereas the duration of the oscillations only depended on the local pattern contrast. We also tested the impulse response before and after the presentation of an adapting stimulus presented in either the same or a different location of the visual field. The response shortened only when both the adapting and the test stimuli were presented at the same location. These last experiments demonstrate a strictly local mechanism of adaptation affecting the response transients of both the impulse and the step response. PMID- 12726836 TI - Adaptation of response transients in fly motion vision. II: Model studies. AB - The so-called 'Reichardt detector' can successfully account for many properties of fly motion vision. In its simplest form, the signals derived from neighboring image locations become multiplied after a low-pass filter has delayed one of them. This operation is done twice in a mirror-symmetrical form and the resulting output signals become finally subtracted. As predicted by this model, fly neurons respond to a brief motion pulse with a sudden rise in activity followed by an exponential decay. The time constant of this decay has been shown to shorten when tested after presentation of an adapting motion stimulus. In terms of the detector model this inevitably implies that the time constant of the low-pass filter is adapting. Given that, one would expect a concomitant shift of the steady-state response towards higher velocities, which, however, could not be experimentally verified. Here, we show that given a model with an additional temporal high-pass filter in the cross-arms of the detector, only the high-pass filter determines the time course of the impulse response. Assuming consequently that the time constant of the high-pass filter is the locus of adaptation resolves the conflicts mentioned above. Moreover, such an elaborated model with an adaptive time-constant faithfully mimics a particular contrast-dependency of transient response oscillations observed in fly motion sensitive neurons. PMID- 12726838 TI - Systematic toxicological analysis: computer-assisted identification of poisons in biological materials. AB - A new software was developed to improve the chances for identification of a "general unknown" in complex biological materials. To achieve this goal, the total ion current chromatogram was simplified by filtering the acquired mass spectra via an automated subtraction procedure, which removed mass spectra originating from the sample matrix, as well as interfering substances from the extraction procedure. It could be shown that this tool emphasizes mass spectra of exceptional compounds, and therefore provides the forensic toxicologist with further evidence-even in cases where mass spectral data of the unknown compound are not available in "standard" spectral libraries. PMID- 12726839 TI - Comparison of a preliminary procedure for the general unknown screening of drugs and toxic compounds using a quadrupole-linear ion-trap mass spectrometer with a liquid chromatography-mass spectrometry reference technique. AB - Liquid chromatography-tandem mass spectrometry (LC-MS-MS) might be a complement to GC-MS and HPLC-diode array detection for the general unknown screening (GUS) of drugs and toxic compounds, particularly when using information- or data dependent acquisition (IDA or DDA), an auto-adaptive MS-MS product-ion scan mode where, at each unit time, the m/z ratios above a given intensity threshold are selected for fragmentation. A new quadrupole-linear ion-trap mass spectrometer (LC-QqQlinear ion-trap) was evaluated for GUS using IDA. For the first detection step (so-called "survey scan") the single quadrupole "enhanced" MS mode (EMS), where ions are accumulated then filtered in the Q3-linear ion-trap, was used. The so-called "enhanced" parent ion scan mode (EPI) used at two alternated fragmentation energies gave the best signal intensity and the best mass spectral information when adding mass spectra obtained in low and high fragmentation conditions, respectively, both in the positive (+20 and +50 eV) and negative (-15 and -40 eV) modes. Solid-phase extracts of serum spiked with eight test compounds (chosen for their retention times distributed along the 30-min long chromatogram and for ionising in both the positive and negative modes) were analysed in parallel with this LC-MS-MS technique and with a reference LC-MS method run on a single-quadrupole instrument where low and high in-source fragmentation conditions in the positive and the negative ion modes are alternated. A C(18), 5 microm (150 x 1 mm I.D.) column and a gradient elution of acetonitrile in pH 3, 2 mM ammonium formate, were used for both. Higher signal-to-noise ratios were obtained with the LC-QqQlinear ion-trap instrument than with the reference technique, resulting in mass spectra devoid of contaminant ions and at least as informative as the reconstructed single-MS spectra. After optimisation of the IDA intensity threshold for the detection of tiny chromatographic peaks in noise, five out of the eight compounds (milrinone, lorazepam, fluometuron, piretanide and warfarin) could be unambiguously identified at the concentration of 0.1 mg/l in serum, in the positive or negative modes, or in both, versus only two by LC MS. All of them could be identified at 1 mg/l by both techniques. These preliminary results show that the sensitivity and mass structural information brought by this new LC-QqQlinear ion-trap instrument may help design an efficient toxicological GUS procedure. PMID- 12726840 TI - Suitability testing of commercial solid-phase extraction sorbents for sample clean-up in systematic toxicological analysis using liquid chromatography (tandem) mass spectrometry. AB - An entire series of SPE sorbents, classified into three different categories (apolar, mixed-mode and polymeric) was evaluated for sample preparation of a data dependent LC-MS-MS "general unknown" screening procedure. An extraction procedure was formulated for each individual column, in agreement with the enclosed instructions, according to the characteristics of each packing. For conciseness, only neutral and basic compounds were chosen for this sorbent suitability test. Thus, the goal of our research was to select the best sorbent with regard to extraction yield and cleanliness of the extracts, all with respect to data dependent acquisition (DDA) mediated LC-MS-MS general unknown screening. We conclude that for that purpose an Isolute C(8) sorbent performs best in terms of extraction yield and clean-up potential. PMID- 12726841 TI - New designer drug p-methoxymethamphetamine: studies on its metabolism and toxicological detection in urine using gas chromatography-mass spectrometry. AB - Studies are described on the metabolism and the toxicological analysis of the new designer drug rac-p-methoxymethamphetamine (PMMA) in rat urine using gas chromatography-mass spectrometry (GC-MS). The identified metabolites indicated that PMMA was extensively metabolized mainly by O-demethylation to pholedrine and to a minor extent to p-methoxyamphetamine (PMA), 1-hydroxypholedrine diastereomers (one being oxilofrine), 4'-hydroxy-3'-methoxymethamphetamine and 4' hydroxy-3'-methoxyamphetamine. The authors' systematic toxicological analysis (STA) procedure using full-scan GC-MS after acid hydrolysis, liquid-liquid extraction and microwave-assisted acetylation allowed the detection of the main metabolites of PMMA in rat urine after a dose corresponding to that of drug users. Therefore, this procedure should be suitable for detection of PMMA intake in human urine via its metabolites. However, it must be considered that pholedrine and oxilofrine are also in therapeutic use. Differentiation of PMMA, PMA and/or pholedrine intake is discussed. PMID- 12726842 TI - Studies on the metabolism and toxicological detection of the Eschscholtzia californica alkaloids californine and protopine in urine using gas chromatography mass spectrometry. AB - Eschscholtzia californica preparations are in use as phytopharmaceuticals and as herbal drugs. Studies are described on the metabolism and the toxicological analysis of the Eschscholtzia californica alkaloids californine and protopine in rat urine using gas chromatography-mass spectrometry. The identified metabolites indicated that californine is extensively metabolized by N-demethylation and/or single or double demethylenation with consecutive catechol-O-methylation of one of the hydroxy groups. Protopine, however, only undergoes extensive demethylenation of the 2,3-methylenedioxy group followed by catechol-O methylation. All phenolic hydroxy metabolites were found to be partly conjugated. The authors' systematic toxicological analysis procedure using full-scan gas chromatography-mass spectrometry after acid hydrolysis, liquid-liquid extraction and microwave-assisted acetylation allowed the detection of the main metabolites of californine and protopine in rat urine after a dose which should correspond to that of drug users. Therefore, use of Eschscholtzia californica preparations should also be detectable in human urine by the authors' systematic toxicological analysis procedure. PMID- 12726843 TI - Determination of amphetamines in human urine by headspace solid-phase microextraction and gas chromatography. AB - Solid-phase microextraction (SPME) is under investigation for its usefulness in the determination of a widening variety of volatile and semivolatile analytes in biological fluids and materials. Semivolatiles are increasingly under study as analytical targets, and difficulties with small partition coefficients and long equilibration times have been identified. Amphetamines were selected as semivolatiles exhibiting these limitations and methods to optimize their determination were investigated. A 100- micro m polydimethylsiloxane (PDMS) coated SPME fiber was used for the extraction of the amphetamines from human urine. Amphetamine determination was made using gas chromatography (GC) with flame-ionization detection (FID). Temperature, time and salt saturation were optimized to obtain consistent extraction. A simple procedure for the analysis of amphetamine (AMP) and methamphetamine (MA) in urine was developed and another for 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxy-N-methamphetamine (MDMA) and 3,4-methylenedioxy-N-ethylamphetamine (MDEA) using headspace solid-phase microextraction (HS-SPME) and GC-FID. Higher recoveries were obtained for amphetamine (19.5-47%) and methamphetamine (20-38.1%) than MDA (5.1-6.6%), MDMA (7-9.6%) and MDEA (5.4-9.6%). PMID- 12726844 TI - Routine analysis of amphetamine and methamphetamine in biological materials by gas chromatography-mass spectrometry and on-column derivatization. AB - A simple determination method of amphetamine (AP) and methamphetamine (MA) in biological materials was developed using on-column derivatization and gas chromatography-mass spectrometry (GC-MS). AP and MA in biological materials were adsorbed on the surface of Extrelut and then extracted and derivatized simultaneously on the Extrelut column. AP and MA were derivatized to the N propoxycarbonyl derivatives using propylchloroformate. Pentadeuterated MA was used as an internal standard. The recoveries of AP and MA from urine were 88.2 and 92.5%, and those from blood were 89.7 and 90.3%, respectively. The calibration curves showed linearity in the range of 12.5-2000 ng/ml (ng/g) for AP and MA in urine and blood, and 0.25-20 ng/mg in hair. When urine samples containing two different concentrations (200 and 1000 ng/ml) of AP and MA, blood samples containing two different concentrations (200 and 1000 ng/g) of AP and MA, hair samples containing two different concentrations (0.5 and 5.0 ng/mg) of AP and MA, the coefficients of variation of intra-day and inter-day were 0.68-3.60% in urine, 0.42-4.58% in blood, and 1.20-13.1% in hair. Furthermore, this proposed method was applied to a medico-legal case of MA intoxication. PMID- 12726845 TI - Solid-phase micro-extraction-gas chromatography-mass spectrometry and headspace gas chromatography of tetrahydrocannabinol, amphetamine, methamphetamine, cocaine and ethanol in saliva samples. AB - In the present work, a method was developed aiming at the serial detection of tetrahydrocannabinol (THC), amphetamine, methamphetamine, cocaine and ethanol in saliva. Saliva samples were submitted to an initial headspace procedure for ethanol determination by gas chromatography/flame ionization detector (GC-FID). After this step, two consecutive solid-phase micro-extractions (SPME) were carried out: THC was extracted by submersing a polydimethylsiloxane fiber (100 micro m) in the vial for 20 min; amphetamine, methamphetamine and cocaine were subsequently extracted after alkalinization. Derivatization of the amphetamines was carried out directly in the solution by adding 2 micro l of butylchloroformate. Gas chromatography-mass spectrometry (GC-MS) was used to identify the analytes in selected ion monitoring (SIM) mode. Confidence parameters of validation of the method were: recovery, linearity, intra- and inter-assay precision as well as limits of detection and quantification of the analytes. The limits of quantification (LOQ) obtained were: ethanol (0.010 g/l); amphetamine (5.0 ng/ml); methamphetamine (0.5 ng/ml); cocaine (5 ng/ml) and THC (5 ng/ml). The method proved to be highly precise (coefficient of variation<8%) for all detected substances. PMID- 12726847 TI - Hair analysis for opiates: evaluation of washing and incubation procedures. AB - Hair analysis of drugs of abuse has been a subject of interest from a clinical, social and forensic perspective for years because of the broad time detection window after intake in comparison to urine or blood. However, the correct and reliable interpretation of opiates findings in an authentic hair sample requires optimalisation and standardisation of decontamination and incubation procedures. Comparing various published methods, we have found some variability in them and no unequivocal recommended procedure for starting with a method directly. Therefore, various combinations of solvents, of various polarity, as washing solvents were tested for removing opiates from the external surface of real hair samples. The yields of opiates from these washings were compared with the yields from the interior of the hair matrix after digestion with various procedures. The opiates after digestion were cleaned up from resulting solution on extraction columns with mixed solid-phase and analysed by GC-MS in standard EI mode after silylation. The efficiencies of neutral (Soerensen buffer, pH 7.4), acid (0.1 M HCl) and basic (1 M NaOH) digestion of the hair matrix were evaluated and the relative recoveries for morphine, codeine, dihydrocodeine and hydrocodone were compared. As it is very problematic to imitate the reference hair sample with a specific amount of analytes incorporated inside, which can be used for calibration to get a close estimate of the quantities of analytes inside the solid authentic sample, the total digestion of a hair sample in basic medium was considered to be a very important reference basis for quantitative determinations. The ratios of hydrolysis of labile 6-acetylmorphine or acetylcodeine were tested and evaluated in practical routine conditions of acid or neutral digestion of hair. Comparing the three methods of incubation of authentic hair samples, the methods using 1 M NaOH or 0.1 M HCl yielded higher recoveries of total equivalents of morphine or codeine, whereas the incubation in Soerensen buffer allowed the reflection of real ratios of labile metabolites and/or parent compounds in an original sample. This method has been shown to be capable of detecting hydrocodone in hair with other opiates concomitantly and to indicate the drug abuse pattern of a person at various time intervals in the past. PMID- 12726846 TI - New designer drug 4'-methyl-alpha-pyrrolidinohexanophenone: studies on its metabolism and toxicological detection in urine using gas chromatography-mass spectrometry. AB - R,S-4'-Methyl-alpha-pyrrolidinohexanophenone (MPHP) is a new designer drug which has appeared on the illicit drug market. The aim of this study was to identify the MPHP metabolites using solid-phase extraction, ethylation or acetylation, as well as to develop a toxicological detection procedure in urine using solid-phase extraction, trimethylsilylation and GC-MS. Analysis of urine samples of rats treated with MPHP revealed that MPHP was completely metabolized by hydroxylation of the tolyl methyl group followed by dehydrogenation to the corresponding carboxylic acid, hydroxylation of the side chain, hydroxylation of the pyrrolidine ring with subsequent dehydrogenation to the corresponding lactam and/or reduction of the keto group. The carboxy and/or hydroxy groups were found to be only partly conjugated. Based on these data, MPHP could be detected in urine via its metabolites by GC-MS using mass chromatography for screening and library search for identification. PMID- 12726848 TI - Rapid high-performance liquid chromatographic measurement of amisulpride in human plasma: application to manage acute intoxication. AB - Amisulpride, a substituted benzamide derivative, is a second-generation (atypical) antipsychotic and is effective as maintenance therapy in patients with schizophrenia. For toxicological purpose, a rapid RP-HPLC assay was developed for the determination of amisulpride in human plasma. A linear response was observed over the concentration range 100-1000 ng/ml. A good accuracy (< or =5%) was achieved for all quality controls, with intra- and inter-day variation coefficients equal or inferior to 4.9%. The lower limit of quantification was 20 ng/ml, without interferences of endogenous components. This rapid method (run time <5 min) was used to monitor eight intoxications involving amisulpride. PMID- 12726849 TI - Screening procedure for eight quaternary nitrogen muscle relaxants in blood by high-performance liquid chromatography-electrospray ionization mass spectrometry. AB - A screening procedure was developed for the identification and the quantification of eight quaternary nitrogen muscle relaxants, including d-tubocurarine, alcuronium, pancuronium, vecuronium, atracurium, mivacurium, rocuronium and mebezonium, in blood samples. The procedure involves ion-pair extraction with methylene chloride at pH 5.4, reversed-phase HPLC separation and electrospray ionization mass spectrometry detection. The procedure was validated in terms of linearity (0.9295 for all the target compounds at 0.1 mg/l). The screening test was found satisfactory and applied in two fatal deaths. In the first case, toxicological investigations on biological fluids collected during the autopsy revealed the presence of vecuronium (1.2 and 0.6 mg/l in blood and urine, respectively) and its desacetylated metabolite, 3 hydroxy-vecuronium (4.4 and 0.7 mg vecuronium equivalent/l in blood and urine, respectively). In the second forensic case, blood analysis showed high levels of mebezonium (6.5 mg/l). The developed procedure was found suitable for forensic investigations. PMID- 12726850 TI - Kinetics of kavain and its metabolites after oral application. AB - Kavain metabolism in humans was the target of this current investigation. In the present study a high-performance liquid chromatographic (HPLC-DAD) assay method for the simultaneous determination of kavain and its main metabolites (p hydroxykavain, p-hydroxy-5,6-dehydrokavain and p-hydroxy-7,8-dihydrokavain) in serum and urine was developed and validated. The metabolites were mainly excreted in the form of their conjugates. All kavain metabolites were detectable in serum and urine, except for p-hydroxy-7,8-dihydrokavain, which was found in urine only. Confirmation of the results and identification of the metabolites were performed by LC-MS or LC-MS-MS. Kinetics of kavain and its metabolites in serum were investigated after administration of a single oral dose (800 mg kavain). Within 1 and 4 h after uptake, the serum concentrations ranged between 40 and 10 ng/ml for kavain, 300 and 125 ng/ml for p-hydroxykavain, 90 and 40 ng/ml for o-desmethyl hydroxy-5,6-dehydrokavain, and 50 and 30 ng/ml for 5,6-dehydrokavain. PMID- 12726851 TI - Determination of zaleplon and zolpidem by liquid chromatography-turbo-ionspray mass spectrometry: application to forensic cases. AB - Zolpidem and zaleplon are two short-acting hypnotic agents used in Europe and in the USA. An atmospheric pressure ionisation liquid chromatography-mass spectrometry (Sciex API 150 EX) method was developed for the determination of zolpidem and zaleplon in whole blood. After single-step liquid-liquid extraction, the hypnotics were separated by gradient-elution with an ammonium formate buffer/acetonitrile eluent on an Inertsil ODS-3 column. Methaqualone was used as internal standard. The recovery was higher than 70% for both hypnotics and the internal standard. The best fit for the calibration curve was achieved, between 1 and 250 ng/ml, with 1/x quadratic regression. Coefficients of intra- and inter assay variation calculated at 5, 25 and 100 ng/ml were less than 10%. The method was successfully applied to forensic cases. PMID- 12726852 TI - Liquid chromatographic-atmospheric pressure chemical ionization mass spectrometric analysis of opiates and metabolites in rat urine after inhalation of opium. AB - To examine the urinary excretion of opiates and their metabolites following inhalation exposure of rats to opium, analytical procedures for the simultaneous determination of the compounds in opium, the vapor derived by the volatilization of opium and the urine of rats exposed to the opium vapor were developed using liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (LC-APCI-MS). Seven compounds were determined in the opium, namely morphine, codeine, thebaine, noscapine, papaverine, meconic acid and meconin. All seven were extracted with 2.5% acetic acid solution and subjected to LC-APCI-MS analysis. The separation was performed on an ODS column in acetonitrile-50 mM ammonium formate buffer (pH 3.0) using a linear gradient program and quantitative analysis was carried out in the selected ion monitoring mode ([M+H](+)). For the analysis of the volatilization of opium, the opium (1 g) was added to a glass pipe, which was then heated at 300 degrees C for 20 min. Negative pressure (air flow-rate; 300 ml/min) was used to draw the vapor through a series of glass wool and methanol traps. The total amount of each compound in the vapor was estimated by measurement of the compounds trapped in the glass wool and methanol. Wister rats (n=3) were exposed to the vapor derived from the volatilization system and the urinary amounts (0-72 h) of the six opiates and metabolites including morphine-3-grucronide (M3G) and morphine-6-grucronide (M6G) were measured after solid-phase extraction. The calibration curves for those compounds in the rat urine were linear over the concentration range 10-500 ng/ml. The recoveries for each analyte from the rat urine sample spiked with standard solution were generally greater than 80%, and the relative standard deviation for the analytical procedure was less than 8% with the exception of meconin. After inhalation exposure of rats to opium, M3G (5.45-14.38 micro g), morphine (2.27 4.65 micro g), meconin (0.54-1.85 micro g), codeine (0.54-1.85 micro g), noscapine (0.34-0.40 micro g) and papaverine (0.01-0.04 micro g) were detected in the urine over 72 h. However, only trace levels of thebaine were observed despite it being one of the major alkaloids found in the opium. On the other hand, a relatively large amount of meconin was detected in the vapor and the urine as compared with the opium. It is suggested that the presence of meconin in biological fluids could be indicative of opium ingestion by inhalation. PMID- 12726853 TI - Evaluation of electrospray ionisation liquid chromatography-tandem mass spectrometry for rational determination of a number of neuroleptics and their major metabolites in human body fluids and tissues. AB - A study of liquid chromatography-triple quadrupole mass spectrometry (LC-MS-MS) with positive electrospray ionisation (ESI) for the determination of selected drugs in human tissues and body fluids such as blood, urine and hair is described. The possibility to screen for and quantify the 19 most commonly prescribed neuroleptics on the Swedish market and determine the presence of their major metabolites within a single LC-MS-MS analysis was evaluated on a PE Sciex API2000 instrument. Chromatographic conditions were optimised and the best separation, with individual retention times for most of the analytes, was obtained on a Zorbax SB-CN column within a 9-min gradient run. The MS-MS fragmentation conditions were optimised for each compound in order to obtain both specific fragments and high signal intensity. Since neuroleptics are a heterogeneous group of compounds, a markedly difference in collision energy needed to achieve fragments of the selected parent ions was seen and the number of fragments achieved varied as well. For sensitive quantification the transition of the most intense fragment of the protonated molecular ion (M+1)(+) was selected for multiple reaction monitoring analysis. More than 70 transitions were finally included in the assay. Detection levels down to the lower ng/ml level were achieved for all analytes, but between analytes more than a 10-fold difference in signal response was seen. By evaluation of extracted ion chromatograms from the analysis of authentic human blood, urine and hair sample the proposed concept for rational drug analysis was found to be both selective and sensitive for the neuroleptics included. A great number of metabolites could be determined in blood, urine and hair as well. A full method validation was not performed since the objective was to evaluate the method design rather than to validate a final method set-up. PMID- 12726854 TI - p53 mutations and resistance to chemotherapy: A stab in the back for p73. AB - p73 is essential for apoptosis induced by many cytotoxic agents, but this function can be blocked by a particular category of p53 mutations that have consequently acquired a gain of function. PMID- 12726855 TI - Cdk2 dethroned as master of S phase entry. AB - The prevailing view of cdk2 as a critical regulator of cell cycle progression and optimal therapeutic target in cancer cells is now challenged by the observation that tumor cells deficient in cdk2 protein and kinase activity are not impaired in proliferation. PMID- 12726856 TI - Connecting estrogen receptor function, transcriptional repression, and E-cadherin expression in breast cancer. AB - A recent paper in Cell (Fujita et al., 2003) demonstrates that MTA3, a novel component of the Mi-2/NuRD transcriptional repression complex, is an estrogen receptor-regulated inhibitor of the Snail zinc finger transcription factor in breast cancer. Given the important role of Snail in repressing E-cadherin transcription and the function of E-cadherin as a tumor suppressor protein and regulator of epithelial architecture, the findings offer potentially significant new insights into cancer pathogenesis. PMID- 12726857 TI - Emerging roles for E2F: beyond the G1/S transition and DNA replication. AB - E2F activity is crucial for the G1/S transition and DNA replication in mammalian cells. The retinoblastoma (pRB) family of proteins is the primary negative regulator of E2F. Recent findings have begun to clarify distinct roles for E2F family members during cell cycle progression and have considerably broadened our understanding of E2F transcriptional control beyond S phase. In this review, we examine the relative contribution of two distinct subclasses of E2F to repression and activation and how this division of labor could explain the role of E2F in DNA damage and repair checkpoints as well as tumorigenesis. PMID- 12726859 TI - In vivo imaging of molecular-genetic targets for cancer therapy. PMID- 12726858 TI - Protein microarrays: meeting analytical challenges for clinical applications. AB - Protein microarrays, one emerging class of proteomic technologies, have broad applications for discovery and quantitative analysis. A rapidly expanding use of this technology is the acquisition of information about the posttranslational modifications of proteins reflecting the activity state of signal pathways and networks, and is now employed for the analysis of biopsy samples in clinical trial research. PMID- 12726861 TI - Hypoxia promotes invasive growth by transcriptional activation of the met protooncogene. AB - Hypoxia unleashes the invasive and metastatic potential of tumor cells by largely unknown mechanisms. The Met tyrosine kinase, a high affinity receptor for hepatocyte growth factor (HGF), plays a crucial role in controlling invasive growth and is often overexpressed in cancer. Here we show that: (1) hypoxia activates transcription of the met protooncogene, resulting in higher levels of Met; (2) hypoxic areas of tumors overexpress Met; (3) hypoxia amplifies HGF signaling; (4) hypoxia synergizes with HGF in inducing invasion; (5) the proinvasive effects of hypoxia are mimicked by Met overexpression; and (6) inhibition of Met expression prevents hypoxia-induced invasive growth. These data show that hypoxia promotes tumor invasion by sensitizing cells to HGF stimulation, providing a molecular basis to explain Met overexpression in cancer. PMID- 12726860 TI - Inactivation of E2F3 results in centrosome amplification. AB - The E2F family of transcription factors is critical for the control of cell cycle progression. We now show that the specific inactivation of E2F3 in mouse embryo fibroblasts (MEFs) results in a disruption of the centrosome duplication cycle. Loss of E2F3, but not E2F1, E2F2, E2F4, or E2F5 results in unregulated cyclin E dependent kinase activity, defects in nucleophosmin B association with centrosomes, and premature centriole separation and duplication. Consequently, this defect leads to centrosome amplification, mitotic spindle defects, and aneuploidy. Our findings implicate the E2F3 transcription factor as an important link that orchestrates DNA and centrosome duplication cycles, ensuring the faithful transmission of genetic material to daughter cells. PMID- 12726862 TI - 2ME2 inhibits tumor growth and angiogenesis by disrupting microtubules and dysregulating HIF. AB - Inhibition of angiogenesis is an important new modality for cancer treatment. 2 methoxyestradiol (2ME2) is a novel antitumor and antiangiogenic agent, currently in clinical trials, whose molecular mechanism of action remains unclear. Herein, we report that 2ME2 inhibits tumor growth and angiogenesis at concentrations that efficiently disrupt tumor microtubules (MTs) in vivo. Mechanistically, we found that 2ME2 downregulates hypoxia-inducible factor-1 (HIF) at the posttranscriptional level and inhibits HIF-1-induced transcriptional activation of VEGF expression. Inhibition of HIF-1 occurs downstream of the 2ME2/tubulin interaction, as disruption of interphase MTs is required for HIF-alpha downregulation. These data establish 2ME2 as a small molecule inhibitor of HIF-1 and provide a mechanistic link between the disruption of the MT cytoskeleton and inhibition of angiogenesis. PMID- 12726863 TI - Remodeling of the extracellular matrix through overexpression of collagen VI contributes to cisplatin resistance in ovarian cancer cells. AB - The mechanisms of drug resistance in cancer are poorly understood. Serial analysis of gene expression (SAGE) profiling of cisplatin-resistant and sensitive cells revealed many differentially expressed genes. Remarkably, many ECM genes were elevated in cisplatin-resistant cells. COL6A3 was one of the most highly upregulated genes, and cultivation of cisplatin-sensitive cells in the presence of collagen VI protein promoted resistance in vitro. Staining of ovarian tumors with collagen VI antibodies confirmed collagen VI expression in vivo and suggested reorganization of the extracellular matrix in the vicinity of the tumor. Furthermore, the presence of collagen VI correlated with tumor grade, an ovarian cancer prognostic factor. These results suggest that tumor cells may directly remodel their microenvironment to increase their survival in the presence of chemotherapeutic drugs. PMID- 12726864 TI - p53 polymorphism influences response in cancer chemotherapy via modulation of p73 dependent apoptosis. AB - Intact p73 function is shown to be an important determinant of cellular sensitivity to anticancer agents. Inhibition of p73 function by dominant-negative proteins or by mutant p53 abrogates apoptosis and cytotoxicity induced by these agents. A polymorphism encoding either arginine (72R) or proline (72P) at codon 72 of p53 influences inhibition of p73 by a range of p53 mutants identified in squamous cancers. Clinical response following cisplatin-based chemo-radiotherapy for advanced head and neck cancer is influenced by this polymorphism, cancers expressing 72R mutants having lower response rates than those expressing 72P mutants. Polymorphism in p53 may influence individual responsiveness to cancer therapy. PMID- 12726865 TI - Chemosensitivity linked to p73 function. AB - Most chemotherapeutic agents induce DNA damage, leading to p53 accumulation and apoptosis. The factors that determine chemosensitivity in p53-defective tumor cells are poorly understood. We found that the p53 family member p73 is induced by a wide variety of chemotherapeutic drugs. Blocking p73 function with a dominant-negative mutant, siRNA, or homologous recombination led to chemoresistance of human tumor cells and engineered transformed cells, irrespective of p53 status. Mutant p53 can inactivate p73 and downregulation of mutant p53 enhanced chemosensitivity. These findings indicate that p73 is a determinant of chemotherapeutic efficacy in humans. PMID- 12726866 TI - Standardized health plan reporting in four areas of preventive health care. AB - PURPOSE: This paper discusses first-year reporting by commercial managed care organizations (MCOs) of new measures in the 2000 Health Plan Employer Data and Information Set (HEDIS). The four measures include screening for chlamydia in young women, controlling blood pressure to <140/90mmHg in patients with hypertension, prescribing appropriate medications for persons with asthma (treatment adherence), and providing counseling to women about managing menopause (survey measure). METHODS: In 2000, some 384 commercial MCOs submitted HEDIS results to the National Committee for Quality Assurance (NCQA). Results of the four new HEDIS measures were linked with audit reports and other health plan data reporting characteristics collected by NCQA. Performance variables were stratified by MCOs' willingness to report their results publicly, size of enrollment, performance on other (non-first year) HEDIS measures, and data collection issues. RESULTS: The mean average performance on the four measures was lowest in chlamydia screening in women (16% for ages 21 to 26 years) and highest for use of appropriate medications for people with asthma (59% for ages 18 to 56 years). The mean average of controlling high blood pressure was 39%. Scores on the management of menopause (MoM) measure ranged from 33.7 (for rating of information) to 72.6 (for exposure to counseling). CONCLUSIONS: The initial MCO baseline rates reported here suggest that much work is needed to improve the quality of care in these areas. Plan characteristics shown to be associated with higher performance on existing HEDIS measures do not predict performance on the new measures. In addition, fewer plans reported on the new measures than on established HEDIS measures. To ensure continued improvement in chlamydia screening in women, controlling high blood pressure, use of appropriate medications for people with asthma, and MoM, incentives for tracking and reporting on these health issues must be explored. PMID- 12726867 TI - Cancer screening estimates for U.S. metropolitan areas. AB - OBJECTIVES: To provide estimates of breast, cervical, and colorectal cancer screening for metropolitan areas in the United States. METHODS: Behavioral Risk Factor Surveillance System (BRFSS) data from 1997 to 1999 were reweighted and analyzed for 69 U.S. metropolitan areas for the receipt of a Papanicolaou (Pap) test (ages > or =18 years); mammography (ages > or =40 years); fecal occult blood testing and sigmoidoscopy (ages > or =50 years). Stratified analyses by demographics were performed for 25 metropolitan areas with populations of > or =1.5 million. RESULTS: Metropolitan estimates ranged from 64.6% to 82.0% for mammography and from 77.2% to 91.7% for Pap tests. There was much greater variability in estimates for colorectal cancer screening, with a 3.6-fold difference in the range of estimates for fecal occult blood testing (9.9% to 35.2%) and a 2.5-fold difference for sigmoidoscopy (17.3% to 43.3%). In the 25 largest areas, prevalence of cancer screening was generally lower for persons with a high school education or less and for those without health insurance. Compared with women aged 50 to 64 years, mammography estimates were lower for women aged 40 to 49 years in 13 of the 25 metropolitan areas. Pap testing was less common among women aged > or =65 years, and colorectal cancer screening was less common for persons aged 50 to 64 years. CONCLUSIONS: Estimates of cancer screening varied substantially across metropolitan areas. Increased efforts to improve cancer screening are needed in many urban areas, especially for colorectal cancer screening. The BRFSS is a useful, inexpensive, and timely resource for providing metropolitan-area cancer screening estimates and may be used in the future to guide local or county-level screening efforts. PMID- 12726868 TI - Farm pesticides: outcomes of a randomized controlled intervention to reduce risks. AB - BACKGROUND: In response to the multiple health risks that farm pesticide applicators experience and the need for controlled trials to evaluate prevention programs, this study tested the effects of a small-group educational intervention designed to increase personal protective equipment (PPE) use and to reduce direct pesticide exposure. DESIGN: A randomized controlled design was used with random selection of participants, random assignment to intervention and control groups, and baseline and postintervention assessments. SETTING/PARTICIPANTS: Four hundred Wisconsin dairy farmers certified to apply pesticides to field crops were recruited to participate over a 1-year evaluation period. INTERVENTION: Three hour educational sessions were conducted with approximately 100 randomly assigned participants. Sessions targeted four educational messages: (1) existing evidence of excess cancers among farmers, (2) simulation of pesticide exposure presented through slide show and description, (3) feedback of self-reported data collected from the farmers reporting on frequency of exposure and gear use, and (4) cognitive behavioral strategies that can be adopted to reduce pesticide hazards. MAIN OUTCOME MEASURES: A change in use of required protective equipment use during application and self-reported dermal exposure were evaluated in the control and intervention groups postintervention. RESULTS: Six-month postintervention analyses showed that an educational intervention had significant effects on the use of gloves and gear during the most recent application and an actual reduction in the total number of pesticides used. However, the intervention did not have a significant impact on achieving full PPE compliance nor in reducing the amount of self-reported dermal pesticide exposure during the most recent application reported by applicators. CONCLUSIONS; This one-time educational intervention successfully increased protective equipment use. However, more-intensive programs are needed to achieve greater reductions in personal pesticide exposure. PMID- 12726869 TI - Can primary care doctors prescribe exercise to improve fitness? The Step Test Exercise Prescription (STEP) project. AB - BACKGROUND: Sedentary lifestyle is associated with adverse health outcomes. Available evidence suggests that, despite positive attitudes toward regular exercise in promoting a healthy lifestyle, few physicians actually prescribe exercise for their patients. Barriers include lack of skills and standard office instruments. Because primary care physicians have regular contact with a large proportion of the population, the impact of preventive health interventions may be great. OBJECTIVES: To determine the effect of an exercise prescription instrument (i.e., Step Test Exercise Prescription [STEP]), compared to usual-care exercise counseling delivered by primary care doctors on fitness and exercise self-efficacy among elderly community-dwelling patients. DESIGN: Randomized controlled trial; baseline assessment and intervention delivery with postintervention follow-up at 3, 6, and 12 months. SETTING: Four large (>5000 active patient files) academic, primary care practices: three in urban settings and one in a rural setting, each with four primary care physicians; two clinics provided the STEP intervention and two provided usual care control. PARTICIPANTS: A total of 284 healthy community-dwelling patients (72 per clinic) aged >65 years were recruited in 1998-1999. INTERVENTION: STEP included exercise counseling and prescription of an exercise training heart rate. MAIN OUTCOME MEASURES: The primary outcome measure was aerobic fitness (VO(2max)). Secondary outcomes included predicted VO(2max) from the STEP test, exercise self-efficacy (ESE), and clinical anthropometric parameters. RESULTS: A total of 241 subjects (131 intervention, 110 control) completed the trial. VO(2max) was significantly increased in the STEP intervention group (11%; 21.3 to 24ml/kg/min) compared to the control group (4%; 22 to 23ml/kg/min) over 6 months (p <0.001), and 14% (21.3 to 24.9ml/kg/min) and 3% (22.1 to 22.8ml/kg/min), respectively, at 12 months (p <0.001). A similar significant increase in ESE (32%; 4.6 vs 6.8) was observed for the STEP group compared to the control group (22%; 4.2 vs 5.4) at 12 months (p < 0.001). Systolic blood pressure decreased 7.3% and body mass index decreased 7.4% in the STEP group, with no significant change in the control group (p <0.05). Exercise counseling time was significantly (p <0.02) longer in the STEP (11.7+/ 3.0 min) compared to the control group (7.1+/-7.0 min), but more (p <0.05) subjects completed > or =80% of available exercise opportunities in the STEP group. CONCLUSIONS: Primary care physicians can improve fitness and exercise confidence of their elderly patients using a tailored exercise prescription (e.g., STEP). Further, STEP appears to maintain benefits to 12 months and may improve exercise adherence. Future study should determine the impact of combining cognitive/behavior change strategies with STEP. PMID- 12726870 TI - Environmental measures of physical activity supports: perception versus reality. AB - BACKGROUND: Perceptions of the environment and physical activity have been associated using survey methods, yet little is known about the validity of environmental surveys. In this study, perceptions of the environment at neighborhood and community levels were assessed (1) to determine validity by comparing respondent perceptions to objective measures and (2) to determine test retest reliability of the survey. METHODS: A telephone survey was administered to a stratified sample of Sumter County, South Carolina adults. Respondents' home addresses were mapped using a geographic information system (GIS) (n =1112). As an indicator of validity, kappa statistics were used to measure agreement between perceptions and objective measures identified at neighborhood and community levels using GIS. A second survey in an independent sample (n=408) assessed test retest reliability. RESULTS: When assessing perceptions of environmental and physical activity in a defined geographic area, validity and reliability for neighborhood survey items were kappa= -0.02 to 0.37 and rho=0.42 to 0.74, and for community survey items were kappa= -0.07 to 0.25 and rho=0.28 to 0.56. CONCLUSIONS: Although causality between perception of access and safety and actual physical activity level cannot be assumed, those meeting national physical activity guidelines or reporting some physical activity demonstrated greatest agreement with access to recreation facilities, while those not meeting the guidelines demonstrated greater agreement with safety of recreation facilities. Factors such as distance and behavior may explain differences in perceptions at neighborhood and community levels. Using local environments with short distances in survey methods improves validity and reliability of results. PMID- 12726871 TI - Self-perception of weight appropriateness in the United States. AB - BACKGROUND; The self-perception of weight appropriateness is an important component of eating and weight-loss behaviors. Self-perceived weight status, however, is not fully explained by objective weight status. OBJECTIVE: To examine the influence of sociodemographic factors on Americans' perceptions of their weight appropriateness, controlling for objective weight status. DESIGN: In the Third National Health and Nutrition Examination Survey, respondents were asked, "Do you consider yourself now to be overweight, underweight, or about the right weight?" Responses to this question were compared with how respondents (n=15,593) would be classified by medical standards given their body mass index (BMI). A proportional odds logistic regression model was used to assess the predictive effects of various sociodemographic factors on weight self-perception. RESULTS: Overall, 27.5% of women and 29.8% of men misclassified their own weight status by medical standards. Of particular note, 38.3% of normal weight women thought they were "overweight," while 32.8% of overweight men thought they were "about the right weight" or "underweight." Multivariate regression analysis revealed that, controlling for BMI, numerous factors-including gender, age, marital status, race, income, and education-were independently associated with the self evaluation of weight status. CONCLUSIONS: The self-perceived appropriateness of weight status varies in highly predictable ways among population-level subgroups, likely reflecting differences in the normative evaluation of bodily weight standards. Such evaluations may assist in the explanation of discrepancies between clinical recommendations based on weight status and actual weight control behaviors, discrepancies that are socially patterned along some of the same subgroupings. PMID- 12726872 TI - Willingness to use instant home HIV tests: data from the California Behavioral Risk Factor Surveillance Survey. AB - OBJECTIVE: Although home collection HIV tests are available in the United States, home HIV tests providing instant results are not available. The objective of this study was to examine willingness to use instant home HIV tests and what test characteristics are most important to individuals. METHODS: Six questions were added to the 1999 California Behavioral Risk Factor Surveillance Survey (BRFSS) (n =2964). This state-representative survey is part of the ongoing, 50-state BRFSS sponsored by the Centers for Disease Control and Prevention. Chi-square tests and logistic regression were used to examine willingness to use instant home HIV tests and preferences for HIV test characteristics. RESULTS: Over one third (37%) of respondents would consider using an instant home HIV test. Respondents willing to use instant home tests were more likely to be Hispanic (odds ratio [OR]=1.55; confidence interval [CI]=1.03-2.34); to have not previously been tested for HIV (OR=1.72; CI=1.20-2.45); to be more likely to plan to be tested in the next 12 months (OR=1.17; CI=1.00-1.37); and to prefer urine sample collection (OR=1.56; CI=1.03-2.37). CONCLUSION: Over one third of respondents in a large, state-representative sample would consider using an instant home HIV test. Hispanics, individuals who planned to be tested in the next 12 months, and individuals who had never been tested were more willing to use instant home tests. PMID- 12726873 TI - Shortage of influenza vaccine in 2000-2001: did it change patient beliefs? AB - OBJECTIVE: To determine which patient beliefs associated with influenza vaccination changed during a vaccine shortage year (2000-2001) from a nonshortage year (1999-2000). METHODS: Elderly patients (n=319), who had been previously interviewed about the 1999-2000 influenza season, were interviewed regarding the 2000-2001 season. Participants were from inner-city and suburban medical practices receiving influenza vaccine supply on time or late in the season. RESULTS: Although vaccination rates did not diminish due to the influenza vaccine shortage (73% in 1999-2000 vs 74% in 2000-2001), some patient attitudes changed. More respondents expressed concerns about the influenza vaccine (3% in 1999-2000 vs 12% in 2000-2001, p<0.0001), and fewer responded that friends thought that they should get the vaccine (75% 1999-2000 vs 66% 2000-2001, p=0.005); that if one person in the household "got the flu" others would also (71% 1999-2000 vs 67% 2000-2001, p<0.0001), and that "a person who does not get the flu shot will get influenza" (36% 1999-2000 vs 30% 2000-2001, p=0.02). CONCLUSIONS: The influenza vaccine shortage resulted in a small increase in concerns about influenza vaccine, even in a population with good access to vaccine and high vaccination rates. Vaccine availability updates should include reassurances of vaccine safety and efficacy. PMID- 12726874 TI - Depression, anxiety, and associated health status in low-income Chinese patients. AB - BACKGROUND: Little is known about the prevalence of mental disorders and the associated impact on health status in low-income immigrant Chinese Americans. METHODS: This cross-sectional study surveyed low-income Chinese patients attending a community health center in Chinatown (New York City) from July through November 2001 to determine the prevalence of depressive and anxiety disorders. Selected modules translated into Chinese from the Primary Care Evaluation of Mental Disorders Patient Health Questionnaire (PHQ), a self administered measure designed to diagnose mental health disorders in primary care, were used. To assess the relationship between mental disorders and health status and to examine convergent validity, mean scores on the Medical Outcomes Study 36-Item Short Form Health Survey (SF-36) were examined according to PHQ diagnoses and calculated severity of depression. RESULTS: A total of 856 patients were approached and data were obtained from 523 (61%) eligible participants. Thirty-two (6.1%) patients met the established criteria for major depressive disorder; 26 (5.0%) patients met the criteria for other depressive disorder; and 24 (4.6%) patients met the criteria for other anxiety disorder. Compared to patients without depression or anxiety, patients with major depressive and other anxiety disorder had significantly lower mean scores on all eight SF-36 subscales, while patients with other depressive disorder had lower mean scores on all subscales except for physical functioning. SF-36 subscale scores tended to decrease with increasing depression severity. CONCLUSIONS: Depression and anxiety were prevalent and demonstrated an adverse impact on health status, as measured by the SF-36. Strategies for screening for mental disorders may vary depending on the needs of the primary care setting and may be accomplished using a general health status measure or a disease-specific approach. PMID- 12726875 TI - Graduate programs: what is their contribution to the training of the public health workforce? AB - OBJECTIVES: To determine characteristics of community health/preventive medicine and community health education graduate programs. METHODS: Forty-eight graduate programs were identified as potential respondents and sent a written survey to complete. RESULTS: Forty programs responded; 82% of programs accredited or applying for accreditation responded. During the 1998-1999 period, 3456 students were enrolled and 773 students graduated from these programs, 91% earning the master of public health (MPH) degree. Two thirds of students were employed while enrolled in these programs. CONCLUSION: Graduate programs train a significant number of public health students, meeting the needs of health professionals with flexible course formats in locations where schools of public health are not available. PMID- 12726876 TI - Condom-use errors and problems: a neglected aspect of studies assessing condom effectiveness. AB - OBJECTIVE: To assess and compare condom-use errors and problems among condom using university males and females. METHODS: A convenience sample of 260 undergraduates was utilized. Males (n=118) and females (n=142) reported using condoms in the past 3 months for at least one episode of sex (penis in the mouth, vagina, or rectum) with a partner of the other sex. A questionnaire assessed 15 errors and problems associated with condom use that could be observed or experienced by females as well as males. RESULTS: About 44% reported lack of condom availability. Errors that could contribute to failure included using sharp instruments to open condom packages (11%), storing condoms in wallets (19%), and not using a new condom when switching from one form of sex to another (83%). Thirty-eight percent reported that condoms were applied after sex had begun, and nearly 14% indicated they removed condoms before sex was concluded. Problems included loss of erection during condom application (15%) or during sex (10%). About 28% reported that condoms had either slipped off or broken. Nearly 19% perceived, at least once, that their condom problems necessitated the use of a new condom. Few differences were observed in errors and problems between males and females. CONCLUSIONS: Findings suggest that condom-use errors and problems may be quite common and that assessment of errors and problems do not necessarily need to be gender specific. Findings also suggest that correcting "user failure" may represent an important challenge in the practice of preventive medicine. PMID- 12726877 TI - Graduate programs in public health: major contributions today; tomorrow we've got to get organized. PMID- 12726879 TI - The current plight of occupational medicine physicians. PMID- 12726878 TI - Fighting Back coalition project. PMID- 12726882 TI - G protein signaling and the molecular basis of antidepressant action. AB - Over the past four decades, a variety of interventions have been used for the treatment of clinical depression and other affective disorders. Several distinct pharmacological compounds show therapeutic efficacy. There are three major classes of antidepressant drugs: monoamine oxidase inhibitors (MAOIs), selective serotonin reuptake inhibitors (SSRIs), and tricyclic compounds. There are also a variety of atypical antidepressant drugs, which defy ready classification. Finally, there is electroconvulsive therapy, ECT. All require chronic (2-3 weeks) treatment to achieve a clinical response. To date, no truly inclusive hypothesis concerning a mechanism of action for these diverse therapies has been formed. This review is intended to give an overview of research concerning G protein signaling and the molecular basis of antidepressant action. In it, the authors attempt to discuss progress that has been made in this arena as well as the possibility that some point (or points) along a G protein signaling cascade represent a molecular target for antidepressant therapy that might lead toward a unifying hypothesis for depression. This review is not designed to address the clinical studies. Furthermore, as it is a relatively short paper, citations to the literature are necessarily selective. The authors apologize in advance to authors whose work we have failed to cite. PMID- 12726883 TI - Melatonin in Chinese medicinal herbs. AB - Melatonin is a highly conserved molecule that not only exists in animals, but also is present in bacteria, unicellular organisms and in plants. Since melatonin is an antioxidant, in plants melatonin was speculated to protect them from intrinsic and environmental oxidative stress. More importantly, melatonin in edible plants inevitably enters animals and human through feed and food. In this study, more than 100 Chinese medicinal herbs were analyzed using the methods of solid phase extraction and HPLC-FD on-line with MS to determine whether melatonin is present in these commonly used herbs. Melatonin was detected in majority of these plants. Sixty-four of them contain melatonin in excess of 10 ng per gram dry mass. Melatonin levels in several herbs are in excess of 1000 ng/g. It is well known that normal average physiological plasma levels of melatonin are only 10-60 pg/mL. These high level-melatonin containing plants are traditionally used to treat diseases which presumably involve free radical damage. The current study provides new information concerning one potentially effective constituent present in a large number of medicinal herbs. The results suggest that these herbs should be reevaluated in reference to their nutritional and medicinal value. PMID- 12726884 TI - A rat cell line derived from DMBA-induced mammary carcinoma. AB - A new cell line, designated UHKBR-01, was successfully established from a 7,12 dimethylbenz[a]anthracene (DMBA)-induced rat mammary tumour. DMBA was administered orally at a dose of 4 mg/ml per rat on the first day of the experiment and thereafter at weekly intervals of same dosage, until the rats have reached a weight of around 150-200 g. The tumours grew rapidly after the injection, and were transplanted into nude mice one the harvest size (2.5 x 2 x 1 mm(3)) was reached, it was transplanted onto nude mice. We have developed a cell line from a portion of the DMBA-induced carcinoma of the nude mice. The UHKBR-01 cell exhibited a slow increase in growth rate during the time of culture and was highly tumourigenic in nude mice. The cells have been grown in culture for over 40 passages. Characterization of the cell line was performed. This included morphology by light and transmission electron microscopy, karyotype, growth rate, tumour antigen expression and xenograft implantation into nude mice. These cells exhibit ultrastructural and immunohistochemical features of epithelial cells of mammary origin. The above analyses also demonstrated that UHKBR-01 cells were oestrogen- and progesterone-receptor positive, in likeness to other established breast cancer cell lines such as MDA-MB-231 and MCF-7. The cell line grows as monolayers of oval-shaped cells with large folded nuclei accompanied by a rich supply of mitochondria. This report describes the first in vitro cell line from transplantable DMBA-induced mammary carcinoma of nude mice, which presents unique characteristics that may prove to be a good experimental model for investigating breast cancer biology. PMID- 12726885 TI - Stress interacts with peripheral cholinesterase inhibitors to cause central nervous system effects. AB - Pyridostigmine bromide (PB), a peripheral cholinesterase inhibitor, has been shown to have central cholinesterase inhibition properties under certain conditions (such as when ingested with other chemical compounds or following a high level of stress). Here we tested if stressing rats, using an intermittent 1 hr tailshock protocol, affected the degree of brain acetylcholinesterase (AChE) inhibition caused by a subsequent single injection of PB (2.0 mg/kg) or neostigmine bromide (NB, 0.32 mg/kg), another peripheral carbamate cholinesterase inhibitor. Stressed rats treated with PB had lower levels of AChE activity in the basal forebrain/striatum, but not in other brain areas. Stressed rats treated with NB did not show basal forebrain/striatum AChE activity changes but did show minor reductions of AChE activity in the cortex and cerebellum. These results confirm that prior stress can change the characteristic actions of certain peripherally acting drugs, thus possibly leading to unexpected central nervous system effects. Possible causes for these effects are discussed. PMID- 12726886 TI - Single eight-hour shift of light-dark cycle increases brain-derived neurotrophic factor protein levels in the rat hippocampus. AB - We previously reported that an eight hour phase advance in the light-dark (LD) cycle increases sleep in rats. Brain-derived neurotrophic factor (BDNF) is suggested to be one of the sleep and circadian regulating factors. We have therefore observed the responses of BDNF protein in the hippocampus, cerebellum and brainstem under conditions of LD change. BDNF protein was quantitatively measured using an ELISA kit. Under an 8-h LD phase advance, the levels of hippocampal BDNF were significantly increased on the day of the phase change, while the levels in the cerebellum and brainstem remained constant. Plasma corticosterone levels were not largely affected. Thus, a single LD shift acutely affects hippocampal BDNF metabolism with no large stress response. PMID- 12726887 TI - Evidence for the involvement of a phospholipase C--protein kinase C signaling pathway in insulin stimulated glucose transport in skeletal muscle. AB - The primary purpose of this investigation was to determine the relationship between phospholipase C (PLC) and diacylglycerol (DAG) sensitive protein kinase C isoforms in insulin signaling in skeletal muscle. Using an in vitro preparation of rat soleus muscle we found that insulin (0.6 nM) stimulated glucose transport was inhibited approximately 20 and 25% by the PKC inhibitor GF109203X and the phospholipase C inhibitor U73122 respectively (p<0.05). The combined effects of these inhibitors were no greater than the inhibitory effects of either compound alone. Western blot analysis revealed that insulin induced a redistribution of PKC beta II from the cytosol to the membrane that was reversed in the presence of GF109203X (1 microM) and U73122 (20 microM). Similarly, U73122 and GF109203X reversed the insulin induced increase in membrane associated phosphorylated (ser 660) PKC beta II. The novel finding of this investigation is that insulin induces an increase in PKC beta II translocation and phosphorylation through a U73122 sensitive pathway in quantatively the most important insulin responsive tissue, skeletal muscle. Furthermore, these results imply that PKC beta II may be one of the DAG sensitive isoforms involved in glucose transport. PMID- 12726888 TI - Vascular effects of caffeic acid phenethyl ester (CAPE) on isolated rat thoracic aorta. AB - This study was aimed to investigate the vascular activity of caffeic acid phenethylester (CAPE), one of the major components of honeybee propolis. Experiments were performed on rat thoracic aortic rings, mounted in an isolated organ bath and connected to an isometric force transducer. The effect of CAPE (0.1-300 microM) was evaluated on tissue pre-contracted with phenylephrine (PE, 1 microM) or with KCl (100 mM). In another set of experiments, tissue was incubated with CAPE (1-100 microM) and responses to PE (0.01-3 microM) or KCl (60 mM) were evaluated. The effect of CAPE on cytosolic Ca(2+) concentration in aortic smooth muscle cells stimulated with PE or KCl was also evaluated. CAPE (0.1-300 microM) caused a concentration-dependent relaxation (pEC(50) 4.99 +/- 0.19; Emax 100.75 +/- 1.65%; n = 4) of tissue pre-contracted with PE that was reduced by endothelium removal or by incubation with N(omega)-nitro-L-arginine methyl ester (L-NAME, 100 microM). CAPE also relaxed KCl-precontracted tissue (pEC(50) 4.40 +/ 0.08; n = 4). CAPE inhibited contractile responses to PE or to KCl, and also inhibited the contractile response to PE obtained in a Ca(2+)-free medium. In addition, CAPE inhibited the increase in cytosolic Ca(2+) concentration triggered by stimulation of aortic smooth muscle cells with PE or KCl. Our results demonstrate a vascular activity for CAPE, that is only partially dependent on nitric oxide. Indeed, at high concentrations, CAPE vasorelaxant effect occurs also in absence of endothelium and it is likely due to an inhibitory effect on calcium movements through cell membranes. PMID- 12726889 TI - Protective effect of aqueous garlic extract against oxidative organ damage in a rat model of thermal injury. AB - Oxygen free radicals have been implicated in mediating various pathological processes including burn-induced organ damage. This study was designed to determine the possible protective effect of aqueous garlic extract against oxidative organ damage distant from the original burn wound. Under ether anaesthesia, rats were subjected to severe skin scald injury covering 30% of total body surface area. Rats were decapitated either 2 h or 24 h after burn injury. Aqueous garlic extract (1 ml/kg) was administered i.p. immediately after burn injury. In the 24-h burn group injection was repeated once more (at 12 hour) following the burn injury. Liver, intestine and lung tissues were taken for the determination of malondialdehyde (MDA) and glutathione (GSH) levels, myeloperoxidase (MPO) activity and protein oxidation (PO). Burn injury caused a significant decrease in GSH level, and significant increases in MDA and PO levels, and MPO activity at post-burn 2 and 24 hours. Since garlic extract reversed these oxidant responses it seems likely that garlic extract protects tissues against oxidative damage. PMID- 12726890 TI - High affinity binding of serum histidine-rich glycoprotein to nickel nitrilotriacetic acid: the application to microquantification. AB - Histidine-rich glycoprotein (HRG) is a serum protein with possible pluripotent activities. In this study, a method for the quantification of rabbit histidine rich glycoprotein (rHRG) was developed based upon the high affinity binding profile of rHRG to nickel-nitrilotriacetic acid (Ni-NTA), an improved chelation agent. When the binding profile of Ni-NTA for whole serum proteins was assessed by Western blotting, Ni-NTA exhibited the binding specificity only to rHRG even after washing with 20 mM imidazole, owing to the unusual amounts of histidine residues in rHRG. In the following experiments, the rHRG immobilized onto a microplate with specific antibody was determined spectrophotometrically with peroxidase-labeled Ni-NTA. This method permitted evaluation of rHRG concentrations ranging from 1.0 to 100 ng/ml, and was actually applicable to the monitoring of rHRG in Resource Q-fractionated serum preparations. Also, the co addition of L-histidine into the incubation mixture significantly diminished the specific binding between rHRG and Ni-NTA. These findings indicate the potential usefulness of this method for the specific measurement of small amounts of rHRG and for understanding the roles of abundant histidine residues in rHRG-metal cation interaction. PMID- 12726891 TI - Effect of neonatal beta-endorphin imprinting on sexual behavior and brain serotonin level in adult rats. AB - A single dose (3 microg) beta-endorphin was administered to newborn female and male rats (hormonal imprinting). In adult age (at 5 months) sexual behavior, steroid hormone binding capacity and brain serotonin content was studied. Females' sexual activity (lordosis quotient) significantly decreased and more animals protested against mounting (ratio of kicking and crying 21/24 vs. 8/24; p < 0.001). Males' sexual activity did not change, however more males were aggressive (4/10 vs. 1/10). Uterine estrogen receptor density significantly increased and affinity decreased. There was no change in the binding capacity of thymic glucocorticoid receptors. In the brain, five regions were studied for serotonin content. There was a gender difference in serotonin level and the intragroup differences were also high. In the endorphin treated males the serotonin level was significantly lower than in the controls. In the endorphin treated females the intragroup scattering has been significantly reduced. Nociceptin content of the cerebrospinal fluid was not changed. The experiments call attention to the possibility of adjustment of sexual and behavioral sphere by the individually different endorphin surge during labor. PMID- 12726893 TI - More focus needed on symptom appraisal. PMID- 12726892 TI - Internalization and recycling of human mu opioid receptors expressed in Sf9 insect cells. AB - Internalization and recycling of G protein-coupled receptors (GPCRs), such as the mu-opioid receptor, largely depend on agonist stimulation. Agonist-promoted internalization of some GPCRs has been shown to mediate receptor desensitization, resensitization, and down-regulation. In this study, we investigated whether different mu opioid agonists displayed different effects in receptor internalization and recycling, the potential mechanisms involved in ohmefentanyl induced internalization process. In transfected Sf9 insect cells expressing 6His tagged wild type mu opioid receptor, exposure to 100 nM ohmefentanyl caused a maximum internalization of the receptor at 30 min and receptors seemed to reappear at the cell membrane after 60 min as determined by radioligand binding assay. Ohmefentanyl-induced human mu opioid receptor internalization was concentration-dependent, with about 40% of the receptors internalized following a 30-min exposure to 1 microM ohmefentanyl. 10 microM morphine and 1 microM DAMGO could also induce about 40% internalization. The antagonist naloxone and pretreatment with pertussis toxin both blocked ohmefentanyl-induced internalization without affecting internalization themselves. Incubation with sucrose 0.45 M significantly inhibited ohmefentanyl-induced internalization of the mu receptor. The removal of agonists ohmefentanyl and morphine resulted in the receptors gradually returning to the cell surface over a 60 min period, while the removal of agonist DAMGO only partly resulted in the receptor recycling. The results of this study suggest that ohmefentanyl-induced internalization of human mu opioid receptor in Sf9 insect cells occurs via Gi/o protein-dependent process that likely involves clathrin-coated pits. In addition, the recycling process displays the differential modes of action of different agonists. PMID- 12726894 TI - A new approach to assess illness behaviour. AB - OBJECTIVE: The aim of this study was to develop a scale to assess the different aspects of illness behaviour and to evaluate the measure in depressive and somatising patients. METHODS: The psychometric properties of the scale were assessed in the following diagnostic groups: patients with major depression (n=36), major depressives with an additional somatisation syndrome (n=40), patients with somatisation syndrome (n=37), and a control group (n=37). After multiple test construction procedures and factor analysis, we developed the Scale for the Assessment of Illness Behaviour (SAIB). RESULTS: The scale included 26 items (originally 58 items) and comprised five factors: (1) verification of diagnoses, (2) expression of symptoms, (3) medication, (4) consequences of illness, and (5) scanning. The intercorrelations between these factors were low confirming the multidimensional structure of illness behaviour. Physicians' rating partially confirmed the factors. All patient groups had elevated scores for illness behaviour. Correlational analyses demonstrated that illness behaviour was closely related to anxiety (such as health worries and phobic anxiety), even after controlling for other factors. Somatisation had medium associations with some aspects of illness behaviour. Symptom Checklist (SCL) results revealed that illness behaviour is not associated with the distress of single symptoms, but with the general number of symptoms. CONCLUSION: A suitable instrument to assess illness behaviour has been developed and should be further analysed in subsequent studies. PMID- 12726895 TI - Assessing illness behaviour: what condition is my condition in? PMID- 12726896 TI - Recurrent abdominal pain in children: evidence from a population-based study that social and familial factors play a major role but not Helicobacter pylori infection. AB - OBJECTIVE: To analyze the relationship between social and familial factors, Helicobacter pylori infection and recurrent abdominal pain (RAP) in children in a population-based cross-sectional study among 1221 preschool children aged 5-8 years. METHODS: H. pylori infection status was determined by 13C-urea breath test (13C-UBT) and information on medical history of the child and on RAP as well as on family demographics was obtained by a standardized questionnaire. RESULTS: Overall, 129 children (11.3%) were infected with H. pylori and 29 children were identified as having RAP within the past 3 months (2.5%). Analysis by multiple logistic regression demonstrated a clear relationship of RAP with living in a single parent household [odds ratio (OR) 2.9, 95% confidence interval (95% CI) 1.2-6.7], with parental history of peptic ulcer (OR 3.7, 95% CI 1.3-10.4) and with parental history of nonulcer gastrointestinal disorders (OR 5.3, 95% CI 2.1 13.2). By contrast, there was a nonsignificant relation between H. pylori infection and occurrence of RAP (OR 1.6, 95% CI 0.5-5.5). CONCLUSION: Social and familial factors play a major role but not H. pylori infection in RAP. PMID- 12726897 TI - Commentary on Bode et al., recurrent abdominal pain in children. PMID- 12726898 TI - Alexithymia and somatisation: quantitative review of the literature. AB - OBJECTIVE: To present a quantitative review of the empirical literature on somatisation and alexithymia. METHODS: Medline and PsycLIT searches for relevant studies were conducted. Meta-analytical techniques were applied to quantify the strength of the associations that were found. RESULTS: A small to moderate relationship was found between general alexithymia and somatic symptom reporting. The alexithymia dimension measuring difficulty in identifying feelings showed the strongest association with symptom reports. The alexithymia dimension measuring externally oriented thinking was virtually unrelated to somatic symptom reports. Compared to healthy control populations, subjects suffering from a somatoform condition were significantly more alexithymic, with effect sizes ranging from moderate to large. The studies comparing somatoform to medical or psychiatric conditions yielded inconclusive results. CONCLUSIONS: By means of quantitative procedures, an association between general alexithymia and somatic symptom reporting was established. Due to the use of questionnaires that can only check for symptoms, not whether these symptoms are medically explained or not, it is however not possible to draw conclusions on somatisation properly defined. The inconsistent results found when comparing somatoform conditions to medical and psychiatric controls may be attributed to confounding variables. In future studies, these variables should be statistically controlled to establish a more consistent pattern of associations between somatoform conditions and alexithymia. It is, however, equally feasible that this inconsistency reflects the nonspecific character of the association between alexithymia and somatisation. The presence of only one prospective study does not allow to draw conclusions on alexithymia as a predisposing factor for somatisation. PMID- 12726899 TI - Alexithymia in somatoform and depressive disorders. AB - OBJECTIVE: Alexithymia and its association with attribution styles, amplification and illness attitudes was studied among subjects with somatoform disorders, depressive disorders and normal subjects. METHODS: Two groups of 30 subjects each, bearing diagnoses of somatoform disorder and depressive disorder respectively (ICD-10 DCR), and one group of 30 normal controls were recruited. The study subjects were assessed using the Toronto Alexithymia Scale and scales for assessing attribution styles, amplification and illness attitudes. RESULTS: Mean alexithymia scores in the somatoform (60.4) and depressive disorder groups (62.5) were higher than in normal subjects (54.2). In the somatoform disorder group, total alexithymia and 'difficulty describing feelings' scores positively correlated with psychological attribution (the latter correlation was also noted in the depressive disorder group), but not with the illness attitudes, amplification, somatic attribution scores or any of the sociodemographic variables. Compared with normal subjects, those with somatoform and depressive disorder had greater difficulty in identifying bodily sensations and feelings. Subjects with depressive disorder had more difficulty in expressing feelings compared to somatoform disorder subjects. CONCLUSIONS: While total alexithymia scores do not differentiate somatoform from depressive disorders, the two diagnostic groups do differ in that depressed subjects have greater difficulty in expressing feelings. However, all three groups had mean scores within the non alexithymic range. Alexithymia and difficulty in expressing feelings were associated with psychological attribution of innocuous bodily sensations in the somatoform disorder group suggesting that alexithymic subjects are more able to psychologize bodily symptoms than non-alexithymic subjects. Somatoform and depressive disorder subjects and normals differ from each other in certain alexithymic characteristics, which could have potential therapeutic implications. PMID- 12726900 TI - Childhood experiences of illness and parenting in adults with chronic fatigue syndrome. AB - OBJECTIVE: There are many similarities between chronic fatigue syndrome (CFS), the somatoform disorders and problems otherwise known as "medically unexplained symptoms." There is some evidence to suggest that a combination of inadequate parenting and early illness experience may predispose the individual to develop medically unexplained symptoms in adult life. The aim of this investigation was to compare the contributions of childhood experiences of illness and parenting in adults with CFS with a fracture clinic control group. METHOD: A retrospective case control design was used. Thirty patients with a diagnosis of CFS and 30 patients attending a fracture clinic in an inner London teaching hospital completed questionnaires measuring parental care and protection and were interviewed about childhood experiences of illness. RESULTS: There were no differences in childhood experience of illness in the two groups. However, logistic regression revealed that maternal overprotection and depression were associated with the diagnosis of CFS. CONCLUSION: The findings may represent risk factors for the development of CFS in adult life. It is possible that maternal overprotection in particular is related to the formation of belief systems about avoiding activity that operate to adversely influence behaviour in patients with CFS. PMID- 12726901 TI - Psychiatric morbidity in the chronic fatigue syndrome: are patients with personality disorder more physically impaired? AB - OBJECTIVE: The long-term consequences of chronic fatigue syndrome (CFS) include substantial impairment in physical functioning and high levels of work disability. In the absence of a medical explanation for this impairment, some have speculated that it may be due to comorbid psychiatric illness or personality disorder. We addressed this possibility by comparing the functional status of three CFS groups: no psychiatric diagnosis, psychiatric illness only, psychiatric illness and personality disorder. A second aim of the study was to determine whether a continuous measure of psychological distress could provide a better account of impairment than psychiatric diagnosis. METHOD: The study sample consisted of 84 consecutive female referrals with CFS. All participants satisfied the case definition and completed an assessment protocol consisting of: physical examination, psychiatric interview and self-report questionnaires. RESULTS: Psychiatric illness, either alone or in combination with a comorbid personality disorder, was not associated with physical impairment or disability in female participants. A regression model of physical functioning found that psychological distress accounted for 6% and symptom severity for 41% of the variance (P=.06 and <.01, respectively). In the case of disability, the corresponding percentages were 2% and 18% (NS and P<.01, respectively). The modest effects of psychological distress could not be attributed to symptom severity. CONCLUSIONS: Although psychiatric illness and personality disorder was prevalent, neither could explain the effects of CFS on physical functioning and disability. As yet, there is no psychological or medical explanation for the behavioral consequences of CFS. PMID- 12726902 TI - Personality traits predict self-referral of young male adults with musculoskeletal complaints to a general practitioner. AB - OBJECTIVE: To investigate the effects of personality traits on self-referral to a general practitioner (GP). METHODS: 217 male soldiers who underwent advanced training course filled the Eysenck Personality Questionnaire-Revised (EPQ-R) and the Perceived Stress Scale and were followed up for a period of 14 weeks. Every soldier who self-referred to the clinic was physically examined by a GP. The major complaint and the GP's diagnosis were documented. RESULTS: 106 of the 217 soldiers self-referred. Self-referred soldiers had significantly (P=.02) higher levels of extraversion than soldiers who did not. Specifically, higher levels of extraversion were evident among soldiers who self-referred with musculoskeletal complaints (P<.05). No significant difference in stress levels was found between soldiers who self-referred and those who did not. CONCLUSIONS: These findings underscore the association between extraversion and musculoskeletal complaints among young adult males and suggest that psychological variables should be taken under consideration in the prevention and treatment of orthopedic problems. PMID- 12726903 TI - Sympathetic reactivity in agoraphobic patients with and without personality disorders. AB - OBJECTIVE: To compare sympathetic activity in agoraphobic patients with and without personality disorders before and after 11 weeks inpatient treatment. METHODS: Agoraphobic patients (n=38), 84% with panic disorder and 47% with personality disorders underwent cold pressure test (CPT), mental stress test (MST), and a specific anxiety test (SAT). Psychological assessments were done by the Bodily Sensations Questionnaire (BSQ), the Agoraphobic Cognitions Questionnaire (ACQ), Spielberger STAI-1 and -2, and a Stress Test Anxiety (STA) questionnaire. Sympathetic activity was measured by blood pressure, heart rate, epinephrine, and norepinephrine. RESULTS: The sympathetic activity did not differ significantly between patient groups, and the reactivity to stress was very low. The sympathetic reactivity remained unchanged after treatment, whereas psychiatric symptoms decreased. Correlations between sympathetic activity and psychological distress were not significant. CONCLUSION: Interpretation of bodily signals seems to be more important than the actual sympathetic activity in agoraphobic patients. PMID- 12726904 TI - Factor analysis and validity of the Transplant Evaluation Rating Scale in a large bone marrow transplant sample. AB - OBJECTIVE: Clinical and methodological challenges are involved in screening bone marrow transplant (BMT) recipients for pretransplant psychosocial adjustment in an attempt to anticipate and prevent behavioral difficulties. Validity of the Transplant Evaluation Rating Scale (TERS), which quantifies the disparate salient elements in a structured clinical assessment, has not been adequately established. This study comprehensively investigated three questions about convergent, internal-structural, and predictive validity of the TERS: how indicative the TERS is of psychosocial difficulties; whether the TERS is uni- or multidimensional; to what degree the TERS predicts long-range adjustment during recovery posttransplant. METHODS: Pre-BMT, 345 consecutive patients were prospectively assessed and completed the MMPI. TERS ratings were assigned retrospectively by two raters (interrater reliability r=.89). RESULTS: The TERS showed good convergent validity relative to MMPI subscales, and a clear, simple, two-factor structure accounting for 47% of the variance. On a subset of our sample (n=29), the factor subscales, "Defiance" and "Emotional Sensitivity," exhibited differential predictive validity to functional status at 1 year posttransplant. CONCLUSIONS: This study, the first large-scale statistical investigation of TERS validity, provided evidence for the validity of the TERS on all three questions. The TERS is indeed indicative of psychosocial risk indexed by MMPI behavioral pathology. It has an understandable, clinically useful factor structure. Its subordinate constructs, Defiance and Emotional Sensitivity, can and should be distinguished conceptually and measured separately. The TERS has clinical utility for specifying behavioral concerns before and guiding proactive intervention after BMT. PMID- 12726905 TI - The effects of vagus nerve stimulation on sleep EEG in depression: a preliminary report. AB - OBJECTIVE: The present study evaluated the effects of vagus nerve stimulation (VNS) on sleep in seven treatment-resistant depressed outpatients. METHODS: Sleep studies were conducted in the laboratory at baseline and 10-12 weeks after VNS implantation while the concomitant psychotropic medication regimen was unchanged. Standard sleep macroarchitecture based on visual stage and assessment of ultradian sleep electroencephalographic (EEG) rhythms were measured on all nights. RESULTS: An overall significant treatment effect on sleep macroarchitecture was obtained by MANOVA. Decreased awake time, decreased Stage 1 sleep and increased Stage 2 sleep were evident post-VNS, although univariate analyses did not reach significance. In addition, the strength or amplitude of ultradian sleep EEG rhythms more than doubled on VNS and was restored to within normal range. CONCLUSION: VNS improved the clinical symptoms of depression and sleep architecture. Results suggest that treatment-resistant depressed patients have dampened sleep EEG rhythms that are restored to near-normal amplitudes with VNS treatment. PMID- 12726906 TI - VoiSS: a patient-derived Voice Symptom Scale. AB - OBJECTIVE: Many voice-rating tools are either physician-derived, disease-specific measures or they merely combine general quality-of-life domains with vocal symptoms. The aim of this series of studies was to devise and validate a patient derived inventory of voice symptoms for use as a sensitive assessment tool of (i) baseline pathology and (ii) response to change in adult dysphonia clinics. METHOD: Three stages in the development of the instrument are described. First, an initial exploratory, open-ended questionnaire study was used to compile a prototype list of voice complaints [Clin Otolaryngol 22 (1997) 37]. Second, the prototype list was administered to 168 subjects with dysphonia and underwent principal components analysis. Qualitatively, it was also assessed at this stage for its ability to capture voice-related impairment, disability and handicap. Third, a modified 44-item scale was administered to 180 new subjects. RESULTS: The symptoms were highly endorsed. Principal components analysis with oblique rotation yielded a Voice Symptom Scale (VoiSS); 43 of the items comprise a 'general voice pathology' scale. More specifically, five oblique components provided assessments of: 'communication problems,' 'throat infections,' 'psychosocial distress,' 'voice sound and variability' and 'phlegm.' CONCLUSION: The VoiSS is simple for patients to complete and easy to score. It is sensitive enough to reflect the wide range of communication, physical symptoms and emotional responses implicit in adult dysphonia. PMID- 12726907 TI - Ability to speak at the age of 1 year and alexithymia 30 years later. AB - OBJECTIVE: We studied the association between speech development in the first year of life and alexithymia in young adulthood. METHODS: The study forms a part of the Northern Finland 1966 Birth Cohort. The original material consisted of all liveborn children in the provinces of Lapland and Oulu in Finland with an expected delivery date during 1966. The comprehensive data collection began during the antenatal phase. In 1997, a 31-year follow-up study was made on a part of the initial sample. The 20-item version of the Toronto Alexithymia Scale (TAS 20) was given to 5983 subjects. Of them, 84% returned the questionnaire properly filled in. The ability to talk was classified according to whether the child spoke no words, one or two words, or three or more words at the age of 1 year. Statistical analyses on the association between the ability to speak at the age of 1 year and alexithymia at the age of 31 years were performed, adjusted for birth weight, mother's parity, place of residence and wantedness of pregnancy. RESULTS: The mean of the total TAS score was lowest among early speakers and for both genders separately. The differences were statistically significant. A parallel significant difference was found among males on TAS Factors 2 and 3 and in case of females on TAS Factors 1 and 3. CONCLUSIONS: We found evidence for an association between speaking development in early childhood and later alexithymia. Our results support the theory that alexithymia may be a developmental process starting in early childhood and reinforcing itself in a social context. PMID- 12726908 TI - Internal consistency of the INTERMED in patients with somatic diseases. PMID- 12726910 TI - Interactions of exogenous or evoked agents and particles: the role of reactive oxygen species. AB - Humans are commonly exposed to combinations of particles (occupational or environmental) and exogenous agents such as ozone and cigarette smoke that generate reactive oxygen species (ROS). Particles also evoke production of ROS from inflammatory cells and of mediators such as TNF-alpha that operate through ROS-related mechanisms. The interactions of particles and ROS-generating agents have been little explored. Adhesion of particles to the surface of pulmonary epithelial cells is increased by exposure to cigarette smoke, ozone, and TNF alpha. Cigarette smoke and ozone increase the uptake of particles by epithelial cells, and both adhesion and uptake can be decreased by scavengers of ROS. Increased adhesion and uptake probably lead to increased levels of inflammatory and fibrogenic mediator production, and cigarette smoke definitely increases whole lung particle retention and enhances the fibrogenic effects of asbestos. In experimental models, the combination of particles plus ozone, cigarette smoke, or reagent hydrogen peroxide augments the inflammatory response to particles, increases cell proliferation, and leads to liberation of increased levels of chemoattractant mediators as well as vascular mediators such as endothelin. The small airways appear to be particular targets of coexposure to smoke or ozone and particles, a phenomenon that may produce chronic airflow obstruction. PMID- 12726911 TI - EPR imaging of reducing activity in Nrf2 transcriptional factor-deficient mice. AB - Mice that lack the Nrf2 (NF-E2-related factor 2) transcription factor develop a lupus-like autoimmune nephritis. The tissue-reducing activity of Nrf2-deficient mice was evaluated using a combination of real-time EPR imaging and spin probe kinetic analysis. Substantial delay in the spin probe 3-carbamoyl-2,2,5,5 tetramethylpyrrolidine-1-oxyl (Carbamoyl-PROXYL) disappearance in the liver and kidneys of Nrf2-deficient mice was observed by EPR imaging. The half-life of the spin probe in the upper abdominal area was prolonged in both the Nrf2-deficient mice and in aged mice. The combination of Nrf2 deficiency and aging in female mice resulted in the most prolonged half-life of disappearance, which was four times longer than that of juvenile female mice with a wild-type genotype. These results indicate that the low reducing activity in these organs is brought about by both Nrf2 deficiency and the aging process, and it may play a key role in the onset of autoimmune nephritis. This combination of the EPR imaging and half-life analysis appears to be a very powerful tool in the real-time analysis of reducing activity. PMID- 12726912 TI - Intralysosomal iron: a major determinant of oxidant-induced cell death. AB - As a result of continuous digestion of iron-containing metalloproteins, the lysosomes within normal cells contain a pool of labile, redox-active, low molecular-weight iron, which may make these organelles particularly susceptible to oxidative damage. Oxidant-mediated destabilization of lysosomal membranes with release of hydrolytic enzymes into the cell cytoplasm can lead to a cascade of events eventuating in cell death (either apoptotic or necrotic depending on the magnitude of the insult). To assess the importance of the intralysosomal pool of redox-active iron, we have temporarily blocked lysosomal digestion by exposing cells to the lysosomotropic alkalinizing agent, ammonium chloride (NH(4)Cl). The consequent increase in lysosomal pH (from ca. 4.5 to > 6) inhibits intralysosomal proteolysis and, hence, the continuous flow of reactive iron into this pool. Preincubation of J774 cells with 10 mM NH(4)Cl for 4 h dramatically decreased apoptotic death caused by subsequent exposure to H(2)O(2), and the protection was as great as that afforded by the powerful iron chelator, desferrioxamine (which probably localizes predominantly in the lysosomal compartment). Sulfide-silver cytochemical detection of iron revealed a pronounced decrease in lysosomal content of redox-active iron after NH(4)Cl exposure, probably due to diminished intralysosomal digestion of iron-containing material coupled with continuing iron export from this organelle. Electron paramagnetic resonance experiments revealed that hydroxyl radical formation, readily detectable in control cells following H(2)O(2) addition, was absent in cells preexposed to 10 mM NH(4)Cl. Thus, the major pool of redox-active, low-molecular-weight iron may be located within the lysosomes. In a number of clinical situations, pharmacologic strategies that minimize the amount or reactivity of intralysosomal iron should be effective in preventing oxidant-induced cell death. PMID- 12726913 TI - Copper induces type II nitric oxide synthase in vivo. AB - Intravenous administration of copper (up to a final concentration of ca. 35 micromol/l in the plasma) led to a progressive, dramatic fall of mean arterial pressure in rats. Copper-induced pressure changes were comparable to those elicited by 2 mg/kg LPS, and were greatly prevented by previous infusion of the inducible NOS (NOS-II) inhibitors aminoguanidine or l-N(6)-(L-imino-ethyl)lysine. RT-PCR analysis showed a significant transcriptional induction of NOS-II in a number of tissues, including aorta, liver, and lungs. Immunohistochemistry revealed that NOS-II was massively synthesized in these tissues upon copper or LPS treatment. The protein was active, as revealed by enzymatic assays on lung homogenates and by the large increase of nitrite/nitrate levels in the plasma. Copper-challenged rats displayed elevated plasma levels of TNFalpha. Extensive formation of nitrotyrosines, indicative of peroxynitrite production, was accompanied by marked morphological changes in examined tissues. Our results clearly show that copper can act as a proinflammatory agent through activation of the nitric oxide pathway, leading to the same pathological frame induced by bacterial lipopolysaccharide. PMID- 12726914 TI - Coxsackievirus B3-resistant mice become susceptible in Se/vitamin E deficiency. AB - The severity of the heart damage caused by a coxsackievirus infection in mice is determined by several factors, including the genotype of the infecting virus as well as the genetic background of the infected host. Earlier work by us showed that the cardiovirulence of a given coxsackievirus genotype could be increased substantially by feeding the host a diet nutritionally deficient in either selenium or vitamin E. Here we report that host genetic background as a determinant of viral infection outcome is superseded by feeding the host a diet nutritionally deficient in both selenium and vitamin E. Mice of the C57Bl/6 strain, normally resistant to coxsackievirus B3-induced myocarditis, become susceptible when fed such a doubly deficient diet. Our results demonstrate the powerful influence of host nutritional status on the course of viral infection compared to other variables traditionally considered to play major roles in determining the extent of virally induced inflammatory heart disease. PMID- 12726915 TI - The phytoestrogen equol increases nitric oxide availability by inhibiting superoxide production: an antioxidant mechanism for cell-mediated LDL modification. AB - Estrogen replacement therapy (ERT) is reported to lower the incidence of cardiovascular disease in postmenopausal women. ERT also lowers the levels of oxidatively modified low-density lipoprotein (LDL). Because modified LDL can mediate the development of atherosclerosis by inflammatory processes, ERT may exert its LDL protective effect through enhanced antioxidant activity in vascular tissues. Plant sources of estrogenic compounds have been used as alternatives for ERT because they avoid a number of negative health effects produced by estrogen. In this study, the antioxidant properties of the soy isoflavone metabolite, equol (an estrogenic metabolite of daidzein) were studied. Equol has a greater antioxidant activity than the parent isoflavone compounds genistein and daidzein, found in high concentration in soy. Equol inhibits LDL oxidation in vitro and LDL oxidative modification by J774 monocyte/macrophages to LDL(-), an electronegative modified LDL found in human plasma. An antioxidant effect of equol was found to be mediated by inhibition of superoxide radical (O(2)(-*)) production and manifested through enhanced levels of free nitric oxide (NO) that prevents LDL modification. Thus, when NO levels were increased by donor agents, generators, or compounds that facilitate nitric oxide synthase activity, LDL(-) formation by J774 cells was strongly inhibited. Conversely, inhibition of NO production enhanced LDL(-) formation, and the combination of reduced NO and increased O(2)( *) production yielded maximum LDL(-) formation. Pretreatment of cells with equol inhibited production of O(2)(-*) by J774 cells apparently via the inactivation of the reduced nicotinamide adenine dinucleotide phosphate oxidase complex. Decreased O(2)(-*) production resulted in increased free NO levels (but not total NO production) indicating that decreased reactions between O(2)(-*) and NO are an outcome of equol's antioxidant activity in cell culture. PMID- 12726916 TI - Glutathione depletion increases nitric oxide-induced oxidative stress in primary rat hepatocyte cultures: involvement of low-molecular-weight iron. AB - Various drugs and chemicals can cause a glutathione (GSH) depletion in the liver. Moreover, nitric oxide (NO) can be generated in response to physiological and pathological situations such as inflammation. The aim of this study was to estimate oxidative stress when primary rat hepatocytes were exposed to GSH depletion after NO production. For this purpose, cells were preincubated with lipopolysaccharide (LPS) and gamma-interferon (IFN) for 18 h in order to induce NO production by NO synthase and then L-buthionine sulfoximine (BSO), an inhibitor of GSH synthesis, was added for 5 h. In hepatocyte cultures preincubated with LPS and IFN before BSO addition, an increase in lipid peroxidation was noted. In those cells, an elevation of iron-bound NO and a decrease in free NO led us to suggest the involvement of low-molecular-weight iron (LMW iron) in the enhancement of oxidative stress. Indeed, addition of deferiprone, a chelator of LMW iron, reduced iron-bound NO levels and the extent of oxidative stress. Moreover, an important elevation of LMW iron levels was also observed. As both, N-acetylcysteine, a GSH precursor, and N(G)-monomethyl-L arginine, a NO synthase inhibitor, totally inhibited the elevation of LMW iron and oxidative stress, a cooperative role could be attributed to NO production and GSH depletion. PMID- 12726917 TI - Prevention of oxidant-induced cell death by lysosomotropic iron chelators. AB - Intralysosomal iron powerfully synergizes oxidant-induced cellular damage. The iron chelator, desferrioxamine (DFO), protects cultured cells against oxidant challenge but pharmacologically effective concentrations of this drug cannot readily be achieved in vivo. DFO localizes almost exclusively within the lysosomes following endocytic uptake, suggesting that truly lysosomotropic chelators might be even more effective. We hypothesized that an amine derivative of alpha-lipoamide (LM), 5-[1,2] dithiolan-3-yl-pentanoic acid (2-dimethylamino ethyl)-amide (alpha-lipoic acid-plus [LAP]; pKa = 8.0), would concentrate via proton trapping within lysosomes, and that the vicinal thiols of the reduced form of this agent would interact with intralysosomal iron, preventing oxidant mediated cell damage. Using a thiol-reactive fluorochrome, we find that reduced LAP does accumulate within the lysosomes of cultured J774 cells. Furthermore, LAP is approximately 1000 and 5000 times more effective than LM and DFO, respectively, in protecting lysosomes against oxidant-induced rupture and in preventing ensuing apoptotic cell death. Suppression of lysosomal accumulation of LAP (by ammonium-mediated lysosomal alkalinization) blocks these protective effects. Electron paramagnetic resonance reveals that the intracellular generation of hydroxyl radical following addition of hydrogen peroxide to J774 cells is totally eliminated by pretreatment with either DFO (1 mM) or LAP (0.2 microM) whereas LM (200 microM) is much less effective. PMID- 12726918 TI - Ascorbate does not act as a pro-oxidant towards lipids and proteins in human plasma exposed to redox-active transition metal ions and hydrogen peroxide. AB - The combination of ascorbate, transition metal ions, and hydrogen peroxide (H(2)O(2)) is an efficient hydroxyl radical generating system called "the Udenfriend system." Although the pro-oxidant role of ascorbate in this system has been well characterized in vitro, it is uncertain whether ascorbate also acts as a pro-oxidant under physiological conditions. To address this question, human plasma, used as a representative biological fluid, was either depleted of endogenous ascorbate with ascorbate oxidase, left untreated, or supplemented with 25 microM-1 mM ascorbate. Subsequently, the plasma samples were incubated at 37 degrees C with 50 microM-1 mM iron (from ferrous ammonium sulfate), 60 or 100 microM copper (from cupric sulfate), and/or 200 microM or 1 mM H(2)O(2). Although endogenous and added ascorbate was depleted rapidly in the presence of transition metal ions and H(2)O(2), no cholesterol ester hydroperoxides or malondialdehyde were formed, i.e., ascorbate protected against, rather than promoted, lipid peroxidation. Conversely, depletion of endogenous ascorbate was sufficient to cause lipid peroxidation, the rate and extent of which were enhanced by the addition of metal ions but not H(2)O(2). Ascorbate also did not enhance protein oxidation in plasma exposed to metal ions and H(2)O(2), as assessed by protein carbonyl formation and depletion of reduced thiols. Interestingly, neither the rate nor the extent of endogenous alpha-tocopherol oxidation in plasma was affected by any of the treatments. Our data show that even in the presence of redox-active iron or copper and H(2)O(2), ascorbate acts as an antioxidant that prevents lipid peroxidation and does not promote protein oxidation in human plasma in vitro. PMID- 12726919 TI - Bcl-2 family members inhibit oxidative stress-induced programmed cell death in Saccharomyces cerevisiae. AB - Selected antiapoptotic genes were expressed in baker's yeast (Saccharomyces cerevisiae) to evaluate cytoprotective effects during oxidative stress. When exposed to treatments resulting in the generation of reactive oxygen species (ROS), including H(2)O(2), menadione, or heat shock, wild-type yeast died and exhibited apoptotic-like characteristics, consistent with previous studies. Yeast strains were generated expressing nematode ced-9, human bcl-2, or chicken bcl-xl genes. These transformants tolerated a range of oxidative stresses, did not display features associated with apoptosis, and remained viable under conditions that were lethal to wild-type yeast. Yeast strains expressing a mutant antiapoptotic gene (bcl-2 deltaalpha 5-6), known to be nonfunctional in mammalian cells, were unable to tolerate any of the ROS-generating insults. These data are the first report showing CED-9 has cytoprotective effects against oxidative stress, and add CED-9 to the list of Bcl-2 protein family members that modulate ROS-mediated programmed cell death. In addition, these data indicate that Bcl-2 family members protect wild-type yeast from physiological stresses. Taken together, these data support the concept of the broad evolutionary conservation and functional similarity of the apoptotic processes in eukaryotic organisms. PMID- 12726920 TI - Alpha-tocopherol downregulates gulonolactone oxidase activity in sturgeon. AB - Gulonolactone oxidase (GLO) is the enzyme responsible for the last step of ascorbic acid biosynthesis. The aim of this study was to investigate the effect of dietary alpha-tocopherol and ascorbic acid on GLO activity in a lower vertebrate, the white sturgeon (Acipenser transmontanus). Both alpha-tocopherol and ascorbic acid modulated renal GLO activity. The increase of dietary levels of alpha-tocopherol and/or ascorbic acid significantly raised the liver concentrations of these two antioxidants and concomitantly lowered kidney's GLO activity. The results suggest that the enzyme of ascorbic acid synthetic pathway responded to the animal's antioxidant status and that its activity was downregulated by alpha-tocopherol. This is the first record of alpha-tocopherol being involved in the regulation of ascorbic acid synthesis. This new observation may provide a hypothesis for the evolutionary loss of GLO expression in teleost fishes. PMID- 12726921 TI - Role of reactive oxygen species and MAPKs in vanadate-induced G(2)/M phase arrest. AB - Cell growth arrest is an important mechanism in maintaining genomic stability and integrity in response to environmental stress. Using the human lung alveolar epithelial cancer cell line A549, we investigated the role of reactive oxygen species (ROS), extracellular signal-regulated protein kinase (ERK), and p38 protein kinase in vanadate-induced cell growth arrest. Exposure of cells to vanadate led to cell growth arrest at the G(2)/M phase and caused upregulation of p21 and phospho-cdc2 and degradation of cdc25C in a time- and dose-dependent manner. Vanadate stimulated mitogen-activated protein kinases (MAPKs) family members, as determined by the phosphorylation of ERK and p38. PD98059, an inhibitor of ERK, and SB202190, an inhibitor of p38, inhibited vanadate-induced cell growth arrest, upregulation of p21 and cdc2, and degradation of cdc25C. In addition to hydroxyl radical ((*)OH) formation, cellular reduction of vanadate generated superoxide radical (O(2)(*)(-)) and hydrogen peroxide (H(2)O(2)), as determined by confocal microscopy using specific dyes. Generation of O(2)(*)(-) and H(2)O(2) was inhibited by specific antioxidant enzymes, superoxide dismutase (SOD) and catalase, respectively. ROS activate ERK and p38, which in turn upregulate p21 and cdc2 and cause degradation of cdc25C, leading to cell growth arrest at the G(2)/M phase. Specific ROS affect different MAPK family members and cell growth regulatory proteins with different potencies. PMID- 12726923 TI - Genes, environment and Oji-Cree type 2 diabetes. AB - The prevalence of type 2 diabetes in Canadian Oji-Cree is among the highest in the world. Our research has uncovered genetic determinants of Oji-Cree type 2 diabetes and related metabolic traits. The most important genetic discovery by far was the private G319S mutation in transcription factor HNF-1alpha, encoded by the HNF1A gene. HNF1A G319S was discovered by candidate gene sequencing and would have been missed using the currently favored strategy of genome-wide scanning. G319S was associated with increased odds of having type 2 diabetes across the whole study sample and in all subgroups, including adolescent Oji-Cree. Furthermore, G319S had specificity and positive predictive value of 97% and 95%, respectively, for developing type 2 diabetes by age 50. The protein bearing the G319S mutation had impaired function in vitro. Sigmoidal modeling showed that each dose of the G319S allele accelerated the median age of diabetes onset by about 7 yr. This approach also showed that environment more strongly accelerated the median age-of-onset of Oji-Cree diabetes onset than did G319S, which could have implications for intervention strategies to reduce the burden of this epidemic. There is also evidence for genetic determination of related metabolic traits in the Oji-Cree. PMID- 12726922 TI - Hydrogen peroxide stimulates tetrahydrobiopterin synthesis through the induction of GTP-cyclohydrolase I and increases nitric oxide synthase activity in vascular endothelial cells. AB - Tetrahydrobiopterin (BH4), which is an essential cofactor for nitric oxide synthase (NOS), is generally accepted as an important molecular target for oxidative stress. This study examined whether hydrogen peroxide (H(2)O(2)), one of the reactive oxygen species (ROS), affects the BH4 level in vascular endothelial cells (ECs). Interestingly, the addition of H(2)O(2) to ECs markedly increased the BH4 level, but not its oxidized forms. The H(2)O(2)-induced increase in the BH4 level was blocked by the inhibitor of GTP-cyclohydrolase I (GTPCH), which is the rate-limiting enzyme of BH4 synthesis. Moreover, H(2)O(2) induced the expression of GTPCH mRNA, and the inhibitors of protein synthesis blocked the H(2)O(2)-induced increase in the BH4 level. The expression of the inducible isoform of NOS (iNOS) was slightly induced by the treatment with H(2)O(2). Additionally, the L-citrulline formation from L-arginine, which is the marker for NO synthesis, was stimulated by the treatment with H(2)O(2), and the H(2)O(2)-induced L-citrulline formation was strongly attenuated by NOS or GTPCH inhibitor. These results suggest that H(2)O(2) induces BH4 synthesis via the induction of GTPCH, and the increased BH4 is coupled with NO production by coinduced iNOS. H(2)O(2) appears to be one of the important signaling molecules to regulate the BH4-NOS system. PMID- 12726924 TI - A homozygous and a heterozygous defect of the winged helix DNA-binding domain of the fork head (FH) gene in tailless and tailed rat siblings: resultant lymphocyte rich thymoma connected with renal tubule amyloid beta (Abeta) deposits. AB - OBJECTIVE: To investigate the cause of thymoma connected with renal amyloid beta (Abeta) overproduction. DESIGN AND METHODS: Pathologic, immune-fluorescence analyses, flow cytometry (FCM) analyses, polymerase chain reaction (PCR) and reverse transcriptase (RT)-PCR were applied for tailless and tailed Lewis/Sea rat siblings at the age of 6 months. RESULTS: A homozygous defect of the winged helix DNA-binding domain of the fork head (FH) gene was detected in the RT-PCR of the tailless rat kidney and liver. Lymphocyte-rich thymoma was found in both rats, but was more prominent in the tailless rat. FCM showed that CD4+ and CD8a+ cells constituted of more than 90% of the thymus lymphocytes in both rats. Abeta deposits in the renal tubules of both rats were shown by positive staining with antihuman Abeta (11-28) antibody (Ab) together with the activation of the kidney presenilin (PSEN) genes. The mRNA level of transthyretin (TTR) was suppressed moderately with age by the homozygous FH1 gene defect. CONCLUSIONS: Both homozygous and heterozygous defects of the DNA-binding domain of the FH gene caused lymphocyte-rich thymoma and renal Abeta overproduction. PMID- 12726925 TI - The effect of VLDL particles on the accuracy of a direct LDL-cholesterol method in type 2 diabetic patients. AB - OBJECTIVE: To assess the accuracy of the direct method LDL-c Plus, in type 2 diabetic patients.LDL-c Plus was measured in 64 consecutive samples of type 2 diabetic patients and compared with betaquantification (BQ), Friedewald's and an alternative formula. METHODS: LDL-c Plus was also measured in the VLDL (d<1.006 Kg/L) fraction of these samples and in total serum and the VLDL fraction of a phenotype III patient, before and after diluting it with saline or VLDL from normolipidemic subjects. RESULTS: LDL-c Plus showed a significant, constant bias (-8.5 +/- 5.6%) against BQ which correlated with VLDL-cholesterol/total triglyceride ratio (r = 0.760, p < 0.0005); bias decreased to zero when the ratio increased. In the VLDL fraction of the diabetic patients and the phenotype III patient LDL-c Plus measured 20.7 +/- 11.6% and 56.2% of the cholesterol, respectively. Dilution with saline did not alter the latter percentage, whereas dilution with normolipidemic VLDL reduced it showing that LDL-c Plus recognized cholesterol-enriched particles in the d<1.006 Kg/L. Friedewald's formula also showed a significant, constant bias (-3.1 +/- 6.4%) against BQ, whereas the alternative formula did not (0.5 +/- 6.1%). Both calculations classified patients better than LDL-c Plus did at NCEP cut-off points. CONCLUSIONS: In type 2 diabetic patients, LDL-c Plus underestimates LDL-c but measures cholesterol associated to IDL particles in the d<1.006 Kg/L fraction. Although LDLc-Plus might be a better cardiovascular risk estimator when well standardized, at the moment, it does not seem to be superior to calculations. PMID- 12726926 TI - Homocysteine and other plasma amino acids in preeclampsia and in pregnancies without complications. AB - OBJECTIVES: To evaluate a). the plasma amino acid changes observed in pregnant women (n = 124) and b). the homocysteine and other amino acid changes in preeclampsic patients (n = 18), and to determine c) whether these changes were also evident in nonpregnant women with a prior history of preeclampsia (n = 18). DESIGN AND METHODS: Case-control study. Plasma total homocysteine (tHcy): HPLC with fluorescence detection, and amino acids (AA): ion exchange chromatography. RESULTS: a). Significantly lower absolute AA values were observed in the pregnant controls for homocysteine, total, essential, and nonessential AA compared with nonpregnant controls. b. In preeclampsia, significantly higher absolute values of tHcy, total, essential and nonessential AA were observed, but relative values referred to total AA were not different. These changes corrected after delivery. CONCLUSIONS: Hyperhomocysteinemia and an increase in most AA levels were observed in preeclampsia. Relative AA values suggested that these changes might be explained by fluctuations in plasma volume. Abnormal AA levels corrected after delivery. PMID- 12726927 TI - Adenosine deaminase and xanthine oxidase activities in bladder washing fluid from patients with bladder cancer: a preliminary study. AB - Activities of adenosine deaminase (AD), and xanthine oxidase (XO) enzymes were measured in bladder washing fluid (BWF) from 37 patients with bladder cancer. The patients were divided into several groups according to their sex; pattern, number, and depth of the tumors; and tumor grade. There was a statistically significant difference in XO activities between the patients having no tumor and papillary tumor (p < 0.002). The differences in XO values between the patients having no tumor and single tumor; and with no tumor and multiple tumors were statistically significant (p < 0.012, p < 0.016 respectively). XO activities were increased in patients with both papillary and multiple tumors compared to tumor free group. Regarding to the depth of tumors, only the differences in XO values between the patients having no tumor and superficial tumor was statistically significant (p < 0.037). XO values of patients in grade1 were higher than the patients having no tumor (p < 0.010). AD activities in patients with multiple and invasive tumor were increased compared to patients with single and superficial tumor. AD values in grade 3 were lower than grade 2. However, we did not find any statistically significant differences in AD activities in all groups. As a conclusion, increased XO activity in BWF might be a potentially important finding as an additional diagnostic biochemical tool for bladder cancer. But we could not say this for AD activity. Further investigations in a larger cohort of patients with bladder cancer are needed to enlighten the possible diagnostic role of XO and AD in BWF. PMID- 12726928 TI - A microarray to analyze methylation patterns of p16(Ink4a) gene 5'-CpG islands. AB - OBJECTIVES: Aberrant DNA methylation of the CpG site is among the earliest and most frequent alterations in cancer. Several studies suggest that aberrant methylation on the CpG sites of the tumor suppressor gene is closely associated with carcinogenesis. However, large-scale analysis of candidate genes has so far been hampered by the lack of high throughput approach for analyzing methylation patterns. The aim of this study was to develop a new method to analyze methylation patterns of p16(Ink4a) gene. DESIGN AND METHODS: We selected a 336 bp segment of the 5' untranslated region and the first exon of the p16(Ink4a) gene, as the target sequence, which include the most densely packed CpG fragment of the islands containing 32 CpG sites. A set of oligonucleotide probes was designed to assemble a DNA microarray to discriminate the methylation patterns of several adjacent CpG sites. RESULTS: Methylation patterns of human p16(Ink4a) gene were mapped and the results were validated by bisulphite DNA sequencing. A good reproducibility was observed in several parallel experiments. CONCLUSIONS: The methylation oligonucleotide microarray can be applied as a useful and powerful tool to map methylation patterns in multiple CpG island sites. PMID- 12726929 TI - Evaluation of newly developed ELISA using "MESACUP-2 test mitochondrial M2" kit for the diagnosis of primary biliary cirrhosis. AB - OBJECTIVES: An enzyme-linked immunosorbent assay (ELISA) using MESACUP-2 Test Mitochondria M2 kit (new-M2 ELISA) has recently become commercially available. The aim of this study was to evaluate the clinical utility of this newly developed ELISA for the diagnosis of primary biliary cirrhosis (PBC). DESIGN AND METHODS: We tested the immunoreactivity of sera from 82 Japanese PBC patients to the 2-oxo-acid dehydrogenase complex (2-OADC) enzymes by indirect immunofluorescence, enzyme inhibition assay using commercially available TRACE Enzymatic Mitochondrial Antibody (M2) Assay (EMA) kit, commercial ELISAs using MESACUP Mitochondria M2 kit (old-M2 ELISA) and new-M2 ELISA, and immunoblotting on bovine heart mitochondria. RESULTS: Each test gave the following positive results; antimitochondrial antibodies (AMA) by immunofluorescence in 71 (87%) out of the 82 sera, enzymatic inhibitory antibody to pyruvate dehydrogenase complex (PDC) by EMA in 61 (74%), immunoglobulin (Ig) G class anti-PDC antibody by old-M2 ELISA in 55 (67%), IgG/M/A class anti-E2 subunit of PDC (PDC-E2)/anti-E2 subunit of branched chain oxo-acid dehydrogenase complex (BCOADC-E2)/anti-E2 subunit of 2 oxoglutarate dehydrogenase complex (OGDC-E2) antibodies by new-M2 ELISA in 73 (89%), and IgG, IgM, or IgA class antibodies against at least one of the 2-OADC enzymes by immunoblotting in 82 (100%). Fifty-three of the 82 sera (65%) were all positive by these five assays. Of the 18 sera that were positive by new-M2 ELISA but negative by old-M2 ELISA, 12 were theoretically interpretable. Of the 11 sera that were negative for AMA by immunofluorescence but positive for at least one of anti-2-OADC enzymes by immunoblotting, four (36%) were positive by new-M2 ELISA, whereas only two and one sera were positive by EMA and old-M2 ELISA, respectively. CONCLUSIONS: Our results indicated that the sensitivity of the newly developed new-M2 ELISA was higher than that of EMA and old-M2 ELISA, and comparable with that of immunofluorescence. However, it is still unclear whether the new-M2 ELISA could replace the conventional immunofluorescence testing for routine assay requests because six (7%) sera showed discrepant results between these two assays. PMID- 12726930 TI - Salivary cortisol on ROCHE Elecsys immunoassay system: pilot biological variation studies. AB - OBJECTIVES: Salivary analysis is a noninvasive alternative that may be more acceptable to patients, especially children. As such, it has the potential for incorporation into comprehensive, dynamic investigations of metabolic dysfunctions - a significant advancement over a single time point serum analysis. In this study, we wanted to determine if the serum cortisol assay on our routine immunoassay analyzer could reliably measure salivary cortisol concentrations. Because of potential fluctuations in salivary concentrations, we included a biologic variation study as a main facet of our preliminary method evaluation. DESIGN AND METHODS: Twenty-eight healthy individuals (12 males, 16 females) volunteered to provide 5 nonconsecutive first morning saliva samples over a two week period. Samples were stored frozen at home until the completion of the study. Following thawing and centrifugation, cortisol was measured in batch mode for each set of participant samples on the ROCHE Elecsys. Biologic variation was determined following removal of outliers. A method comparison was performed with the DPC Coat-A-Count Cortisol assay following the recommended modifications for salivary analysis, and with the Salimetrics HS-Cortisol two-site monoclonal assay optimized for salivary cortisol. RESULTS: Mean salivary cortisol concentration was 20.4 nmol/L. Analytical variation (CV(A) = 3.8%), within-subject variation (CV(I) = 6.3%), between-subject variation (CV(G) = 20.5%), index of individuality (II = 0.36) and reference change value (RCV = 20.4%) were determined. A negative 40% proportional bias was observed on the Elecsys compared to the two methods that have already been optimized for salivary analysis. CONCLUSIONS: This study demonstrated that salivary cortisol can be reliably measured on a routine automated immunoassay analyzer such as the ROCHE Elecsys. This particular assay needs to be optimized at the low end of the standard curve for routine use with salivary samples. Based on the relatively small intra-individual variation and low index of individuality, reference change values are preferable to the use of population reference intervals for this assay. PMID- 12726931 TI - Detection and quantification of 5-hydroxyoxindole in mammalian sera and tissues by high performance liquid chromatography with multi-electrode electrochemical detection. AB - OBJECTIVE: Since 5-hydroxyoxindole structurally related indole metabolites play different roles in some hepatic and neurologic disorders we found necessary to develop an assay to further investigate the physiologic relevance of this compound. METHODS: We have designed a convenient assay to determine 5 hydroxyoxindole in serum using solid phase extraction and a highly selective High Performance Liquid Chromatography system with multi-Electro Chemical Detection (HPLC-ECD). RESULTS: We have identified and quantified 5-hydroxyoxindole in various mammalian species. Its distribution in tissues showed that the molecule is also present in brain, liver, kidney and spleen, but not in skeletal muscle. CONCLUSIONS: 5-hydroxyoxindole is an endogenous tryptophan metabolite present in circulating blood and in some tissues at the nmol level, its determination using HPLC-ECD will be useful for elucidating the role of this molecule in normal and disease conditions. PMID- 12726932 TI - Serum IGF-I and IGFBP-3 in healthy pregnancies and patients with preeclampsia. AB - DESIGN AND METHODS: We studied serum levels of IGF-I and IGFBP-3 in 68 pregnant women; 28 mild preeclampsia, 13 severe preeclampsia, and 27 healthy pregnant control subjects. RESULTS: When compared with the healthy pregnant group preeclampsia groups had markedly decreased mean serum IGF-I levels. The difference was also significant between the mild and severe preeclampsia groups. Serum mean IGFBP-3 level was also lower in both preeclampsia groups than in healthy pregnancy group, but the difference between the preeclampsia groups did not reach significance. For IGF-I/IGFBP-3 ratio, the mean levels were significantly lower in mild and severe preeclampsia groups than in healthy pregnancy group. Also the mean IGF-I/IGFBP-3 ratio was significantly lower in the severe preeclampsia group compared with the mild preeclampsia group. CONCLUSIONS: IGF-1 and IGFBP-3 levels in patients with preeclampsia were compared with healthy pregnant control subjects. IGF-I and IGF-I/IGFBP-3 ratio may be useful for estimate the severity of preeclampsia. PMID- 12726933 TI - Vascular endothelial growth factor and oxidative damage in cancer. AB - OBJECTIVES: VEGF may be an indicator for the angiogenic potential of a tumor and stimulates NO which plays complex roles in cancer. In our study, we investigated the levels of MDA, NO and VEGF in the plasma of various types cancer patients (untreated, yet). DESIGN AND METHODS: The level of VEGF was determined by using ELISA. Plasma MDA, NO and VEGF levels were measured in 45 patients with various cancer types and in 21 healthy subjects. RESULTS: Plasma MDA and VEGF levels were significantly higher than those of the healthy subjects (p < 0,0001). NO levels of the patients were significantly lower vs. the healthy subjects (p < 0,001). CONCLUSIONS: Increased plasma VEGF and MDA concentrations and decreased plasma NO levels have been found in patients with various types of human cancer. Howewer, the prognostic and clinical significance of plasma VEGF in cancer patients is unknown. PMID- 12726934 TI - False-positive microhematuria in dipsticks urinalysis caused by the presence of semen in urine. AB - OBJECTIVES: Positive readings of blood in dipstick urinalysis may indicate trauma or imbalance in hemostasis associated with drug treatment. We evaluated the possibility that the presence of semen in urine may cause false-positive hematuria. DESIGN AND METHODS: Semen specimens obtained from 25 healthy men were directly applied on urine dipsticks for evaluation of the presence of blood. Isolated sperm cells and seminal fluid were also tested. Dipstick analyses were further performed with semen samples diluted in normal urine. Four healthy male volunteers provided urine samples before and immediately after having had sexual relationships. These samples were dipstick tested for the presence of blood. RESULTS: Semen, spermatozoa and seminal fluid gave false positive results for microhematuria following direct application of samples on dipsticks as well as after their suspension in urine (p < 0.00001).Three out of four postcoital urine specimens yielded positive results for blood. CONCLUSIONS: In men, postcoital urine may be falsely "positive" for microhematuria. This may have implication on the management of male patients in emergency situations such as acute coronary syndromes. PMID- 12726935 TI - Design and construction of a low frequency wide band non-resonant transducer. AB - Commercially available low frequency transducers often have a bandwidth of much less than 100%. This paper reviews the current technology for low frequency transducer construction and presents a non-resonant design giving an improved low frequency performance. Small, undamped piezo-electric cubic elements of 2 mm side length are used which operate below their first resonance frequency. In construction, the elements are air-backed and the front faces are bonded to a thin membrane across the end of an enclosure of 15 mm diameter. Using a low frequency excitation signal which has little energy at the first resonance frequency of the elements, an operating frequency range of 200-500 kHz can be achieved. A single element behaves virtually as a point source, but by arranging an array of elements over the membrane, a more collimated beam profile is obtained. PMID- 12726936 TI - Wideband high-frequency line-focus PVDF transducer for materials characterization. AB - This paper presents the design, fabrication, operating characteristics and applications of a wideband, high-frequency, line-focus beam transducer we constructed using a 9 microm thick piezoelectric polyvinylidene fluoride (PVDF) film. This transducer possesses a focal length of 2.38 mm and an aperture angle of 84 degrees. The frequency spectrum of the signal measured at the focal point indicates that the transducer has a wide frequency response which extends from 10 MHz to over 100 MHz. When compensated for the frequency-dependent attenuation of the coupling medium, the operational frequency exceeds 150 MHz. The transducer can be operated in a time-resolved pulse mode or in a radio-frequency (rf) tone burst mode. An application of the transducer to determine the anisotropic elastic property of a silicon wafer is demonstrated. The phase velocities of surface acoustic waves (SAW) propagating along various directions on the (001) surface of cubic silicon are measured and compared to computed values. PMID- 12726937 TI - A conical air-coupled capacitance transducer for surface imaging. AB - This paper describes the construction and operation of an air-coupled capacitance transducer with a conical backplate. This was designed to produce a focal region over an extended distance along the transducer axis. Measurements were performed to examine both the frequency response of the transducer in pulse-echo mode, and the lateral resolution for imaging purposes. The radiated field was measured and compared to theory, and the extent of the focal region determined. Images of surface topography are presented, to illustrate the range of application of the transducer. PMID- 12726938 TI - Guided wave dispersion curves for a bar with an arbitrary cross-section, a rod and rail example. AB - Theoretical and experimental issues of acquiring dispersion curves for bars of arbitrary cross-section are discussed. Since a guided wave can propagate over long distances in a structure, guided waves have great potential for being applied to the rapid non-destructive evaluation of large structures such as rails in the railroad industry. Such fundamental data as phase velocity, group velocity, and wave structure for each guided wave mode is presented for structures with complicated cross-sectional geometries as rail. Phase velocity and group velocity dispersion curves are obtained for bars with an arbitrary cross-section using a semi-analytical finite element method. Since a large number of propagating modes with close phase velocities exist, dispersion curves consisting of only dominant modes are obtained by calculating the displacement at a received point for each mode. These theoretical dispersion curves agree in characteristic parts with the experimental dispersion curves obtained by a two dimensional Fourier transform technique. PMID- 12726939 TI - Ultrasound-enhanced membrane-cleaning processes applied water treatments: influence of sonic frequency on filtration treatments. AB - Ultrasound (US) cleaning technique was applied to remove fouling of ultrafiltration (UF) and microfiltration (MF) membranes which were used to treat peptone and milk aqueous solutions, respectively. Membrane operations were performed by cross-flow filtration with 60 kPa operating pressure in an US field. The US employed had 28, 45 and 100 kHz frequency with 23 W/cm(2) output power. For each polymeric membrane made of polysulfone UF and cellulose MF, cleaning experiments were carried out with and without US after fouling. The fouled UF and MF membranes showed volume flux decline, but the membrane property was recovered by US irradiation. It was found in 28 kHz frequency that water cleaning was effective for recovery of declined condition due to fouling. Also, US-enhanced permeability of membranes was discussed in both membrane systems. We observed that US decreased the fouling condition in both membrane systems when US was irradiated before fouling. It was found that 28 kHz frequency US could enhance formation of the fouled layer in both filtration systems of peptone and milk solution. PMID- 12726940 TI - The influence of the thickness of non-piezoelectric pieces on pre-stressed piezotransducers. AB - The mechanical pre-stress applied in piezotransducers used to generate high power ultrasound is needed to avoid ceramics fracture on traction cycle. Pre-stress levels inferior to 50 MPa can yield resonance shifting due to effectiveness of acoustic coupling between transducer pieces. Symmetrical transducers with different thickness of passive parts were submitted to axial mechanical pre stress up to 50 MPa and their resonances were measured. The experimental results show the increasing of the resonances frequencies with the level of applied pre stress. Similar effect is verified in simulations by using a model based on Mason's equivalent electric circuit. Due to the similarity of these effects, a relation between applied pre-stress and pieces coupling was proposed for the transducer assembled. In addition, the dependence of the thickness of non piezoelectric pieces on the coupling effectiveness between them is discussed. The results show that transducers with small thickness present more expressive shifting resonance ratio. PMID- 12726941 TI - Anomalous absorption of bulk shear sagittal acoustic waves in a layered structure with viscous fluid. AB - It is demonstrated theoretically that the absorptivity of bulk shear sagittal waves by an ultra-thin layer of viscous fluid between two different elastic media has a strong maximum (in some cases as good as 100%) at an optimal layer thickness. This thickness is usually much smaller than the penetration depths and lengths of transverse and longitudinal waves in the fluid. The angular dependencies of the absorptivity are demonstrated to have significant and unusual structure near critical angles of incidence. The effect of non-Newtonian properties and non-uniformities of the fluid layer on the absorptivity is also investigated. In particular, it is shown that the absorption in a thin layer of viscous fluid is much more sensitive to non-zero relaxation time(s) in the fluid layer than the absorption at an isolated solid-fluid interface. PMID- 12726942 TI - Normal modes of a poroelastic plate and their relation to the reflection and transmission coefficients. AB - A poroelastic plate that obeys the Biot theory is considered. Compact new forms of its reflection and transmission coefficients, similar to those of the resonance scattering theory for an elastic plate, are derived. A numerical comparison of the reflection coefficient modulus with the plate normal modes, at low frequency, shows that a study of the reflection or transmission coefficient does not provide the same kind of information on the poroelastic plate than an investigation of guided leaky waves propagation. PMID- 12726943 TI - A self-aligning ultrasound sensor for detecting foreign bodies in glass containers. AB - This paper introduces the principle and design of an ultrasonic transducer system with auto-alignment mechanism. The proposed system is used for detecting foreign bodies (FBs) in beverage containers. Variation in reflection amplitude is analyzed as a function of the ultrasound beam incident angle to beverage container surface. It is concluded that there exists a quadratic relationship between the strength of the reflected signal and the incident angle. Furthermore, a calculation for effective angular increment for searching the normal of a curved surface is introduced. Experiments conducted using the prototype demonstrated that FBs are detectable in various juices. Experiments also observed some false echo signals that occur due to curved container surface. Threshold in confined time region is therefore set to ensure no fail of detection in low signal to noise ratio. This design is also applicable to non-destructive inspection for metal canned food. PMID- 12726944 TI - Ultrasonic transducers using electron-irradiated vinylidene fluoride trifluoroethylene copolymers. AB - Single-element, planar transducers have been fabricated using electron-irradiated poly(vinylidene fluoride-trifluoroethylene) 80/20 mol% copolymers with different electron dosage. Electrical field-induced strain response of copolymer film with 100 Mrad dosage has been studied at 5 kHz and the electrostrictive coefficient was calculated. The transmitting response of the air-backing and epoxy-backing transducers was evaluated with the application of high DC bias voltages. Clear ultrasonic amplitudes and high frequency spectrum (>20 MHz) were observed when driven from a standard ultrasonic voltage source through a decoupling circuit. It has also showed that larger generation of ultrasonic waves will be induced under high DC bias field, which is due to the increase of induced d(33) piezoelectric coefficient. Two different polar bias voltages, positive and negative, were applied to the transducers and inverse waveforms were received, which was coincident with the theoretical analysis of the strain response of electrostrictive film. PMID- 12726945 TI - A concise and efficient scattering matrix formalism for stable analysis of elastic wave propagation in multilayered anisotropic solids. AB - This paper presents a concise and efficient scattering matrix formalism for stable analysis of elastic wave propagation in multilayered anisotropic solids. The formalism is capable of resolving completely the numerical instability problems associated with transfer matrix method, thereby obviating the extensive reformulation in its modified versions based on delta operator technique. In contrast to the earlier reflection matrix formalisms, all scattering matrices are obtained in a direct manner without invoking wave-propagator or scatterer operator concepts. Both local and global reflection and transmission matrices corresponding to scatterings in two and more layers are derived. The derivation of global scattering matrices in terms of the local ones is carried out concisely based on physical arguments to provide better insights into scattering mechanism. Another formulation which is even more succinct is also devised for obtaining the global scattering matrices directly from eigensolutions. The resultant expressions and algorithm are terse, efficient and convenient for implementation. PMID- 12726946 TI - An efficient synthesis of 3,4-dihydropyrimidin-2-ones catalyzed by NH2SO3H under ultrasound irradiation. AB - The condensation of aldehydes, beta-keto esters and urea catalyzed by NH(2)SO(3)H in ethanol results dihydropyrimidinones in high yields under ultrasound irradiation. PMID- 12726947 TI - Ultrasound promoted synthesis of 2-aroyl-1,3,5-triaryl-4-carbethoxy-4 cyanocyclohexanols. AB - Synthesis of 2-aroyl-1,3,5-triaryl-4-carbethoxy-4-cyanocyclohexanols from chalcones with ethyl cyanoacetate is carried out in excellent yields with KF/basic alumina as catalyst under ultrasound irradiation. PMID- 12726948 TI - Ultrasonic extraction of plant materials--investigation of hemicellulose release from buckwheat hulls. AB - Various one- and two-step extraction procedures with and without a short application of ultrasound at the beginning of the extraction were used to examine the effect of sonication on the extractibility of the hemicellulose components of buckwheat hulls. The polysaccharides recovered from the extracts were characterised by yield as well as composition determined by chemical methods and spectroscopic techniques. They comprised a complex of glucuronoxylan and co extracted amylose-rich starch in various proportions contaminated with other cell wall components (protein, pectic polysaccharides). The hemicellulose fractions obtained by classical and ultrasound-assisted extraction exhibited significant immunomodulatory activities. The increased yield of ultrasonically extracted hemicelluloses, which have preserved their structural and molecular properties as well as immunological activity, confirmed the importance and great potential of ultrasound-assisted extraction of industrially important polysaccharides from different tissues of plant materials. PMID- 12726949 TI - Synergism of ultrasound and solid acids in intensification of Friedel-Crafts acylation of 2-methoxynaphthalene with acetic anhydride. AB - The Friedel-Crafts acylation of 2-methoxynaphthol is generally carried out by using highly polluting acids such as HF, AlCl(3), BF(3) which are used in more than stoichiometric quantities and are neutralized at the end creating large quantities of waste, corrosion problems and hazard. This paper discusses the use of acid treated clays such as K-10 and Filtrol-24 and cation exchange resins such as Amberlyst-36, Amberlyst-15 and Indion-130 as catalysts at 25 degrees C which have different acid strengths and pore size distributions. The activities were in the following order:Amberlyst-36>Indion-130>Amberlyst-15>K-10clay>Filtrol-24. In the presence of ultrasound, the activities were found to increase by more than a factor of 3 in the case of large porous resins than the clays. The selectivity to 1-acyl-2-methoxynaphthalene was found to remain the same. Utrasonics did not promote isomerisation or direct conversion to 6-acyl-2-methoxynaphthalene. PMID- 12726950 TI - Kinetics and mechanisms of the sonolytic destruction of non-volatile organic compounds: investigation of the sonochemical reaction zone using several OH* monitoring techniques. AB - This study investigates the sonolytic degradation mechanism of non-volatile organic compounds and reaction sites for its degradation using various tools that allow OH* to be monitored, such as: the spin-trapping method of OH* detection using non-volatile nitrone trap 5,5-dimethyl-1-pyrroline-N-oxide (DMPO), the hydrogen peroxide analytical methods and the p-chlorobenzoic acid (pCBA)-probe method. These methods can successfully monitor OH* produced during sonochemical processes, and identify the major reaction sites involving OH* of the three proposed reaction zones--within the cavity, in the bulk solution, and at the gas liquid interfacial (shell) region. The patterns of hydrogen peroxide accumulation under the various conditions suggest that peroxides pre-form at the interfacial region, but the self-scavenging reaction by hydrogen peroxide simultaneously takes place in the same region. The simultaneously measured peroxide concentration, in the absence and presence of DMPO, and that of the DMPO-OH adduct indicated the peroxide production and DMPO-OH adduct formation reaction occur at the shell region. The sonolytic destruction efficiency of ultrasound coupled with Fe(II) has been also investigated. The coupled Fe(II)/ultrasound process was found to enhance the OH* production rate by 70% compared to the ultrasound process alone due to the reaction of Fe(II) with sonochemically produced hydrogen peroxide (Fenton's reaction). This accelerated reaction was also found to occur at the shell region rather than in the bulk solution. The enhancement effect of Fe(II)/ultrasound was also examined using pCBA as a probe. 2.8-fold and 3.6-fold increases of the pCBA degradation rate were observed at Fe(II) concentrations of 10 and 20 microM, respectively. PMID- 12726951 TI - A standard method to calibrate sonochemical efficiency of an individual reaction system. AB - Fricke reaction, KI oxidation and decomposition of porphyrin derivatives by use of seven types of sonochemical apparatus in four different laboratories were examined in the range of frequency of 19.5 kHz to 1.2 MHz. The ultrasonic energy dissipated into an apparatus was determined also by calorimetry. Sonochemical efficiency of Fricke reaction and KI oxidation was defined as the number of reacted molecule per unit ultrasonic energy. The sonochemical efficiency is independent of experimental conditions such as the shape of sample cell and irradiation instruments, but depends on the ultrasonic frequency. We propose the KI oxidation dosimetry using 0.1 moldm(-3) KI solution as a standard method to calibrate the sonochemical efficiency of an individual reaction system. PMID- 12726952 TI - Kinetic model for the sonochemical degradation of monocyclic aromatic compounds in aqueous solution: new insights. AB - The kinetic model of De Visscher et al. [J. Phys. Chem. 100 (1996) 11636] for the sonochemical degradation of apolar monocyclic aromatic compounds in aqueous solution is based on a large number of assumptions, some of which have never been stated explicitly. An example is a homogeneous temperature of cavitation. The present study focuses on those "hidden assumptions", and reveals that some of them are not realistic. It is shown that an entirely different set of assumptions leads to essentially the same mathematical formulation of the kinetics. The old and new version of the model can be considered as limiting cases which envelope the real conditions of ultrasonic cavitation chemistry. However, the new model does not lead to the inconsistencies experienced with the old model. PMID- 12726953 TI - OH/D A2sigma(+)-X2pi(i) rovibronic transitions in multibubble sonoluminescence. AB - Multibubble sonoluminescence spectra were recorded in the 300-350 nm wavelength range in the case of H(2)O/Ar, D(2)O/Ar and H(2)O/Kr solutions (acoustic frequency: 20 kHz; spectral resolution optimized to 0.34 nm). Three groups of rotational components (R(1)/R(2), Q(1)/Q(2) and P(1)/P(2)) were identified in the OH/D A2sigma(+)-X2pi(i) (0,0) transitions via the substitution of H(2)O for D(2)O. The congestion of bands and the origin of a red shading extending up to 350 nm are broached. PMID- 12726954 TI - An appropriate response to SARS. PMID- 12726955 TI - Enfuvirtide approved for defusing HIV. PMID- 12726956 TI - Chemical and bio-warfare detection system piloted at US airport. PMID- 12726957 TI - Second case of doctor-to-patient HIV transmission. PMID- 12726958 TI - HIV transmission shift in Europe. PMID- 12726960 TI - Antibody cocktail could cut risk of food poisoning. PMID- 12726961 TI - Meningitis A conjugate vaccine comes on stream. PMID- 12726962 TI - Attack on highland malaria in east Africa. PMID- 12726963 TI - Avian influenza outbreak linked to eye infections. PMID- 12726964 TI - Poliovirus cases rise in India. PMID- 12726966 TI - India makes poor tuberculosis progress in 2002. PMID- 12726967 TI - Haiti's HIV equity initiative. PMID- 12726969 TI - Closing in on the cause of SARS. PMID- 12726970 TI - Revolutionising hand hygiene in health-care settings: guidelines revisited. PMID- 12726971 TI - Revealing all. PMID- 12726972 TI - Epstein-Barr virus reactivation. PMID- 12726973 TI - Efficacy of polysaccharide pneumococcal vaccine in adults in more developed countries: another view of the evidence. PMID- 12726975 TI - Effect of washing hands with soap on diarrhoea risk in the community: a systematic review. AB - We set out to determine the impact of washing hands with soap on the risk of diarrhoeal diseases in the community with a systematic review with random effects meta-analysis. Our data sources were studies linking handwashing with diarrhoeal diseases. Seven intervention studies, six case-control, two cross-sectional, and two cohort studies were located from electronic databases, hand searching, and the authors' collections. The pooled relative risk of diarrhoeal disease associated with not washing hands from the intervention trials was 1.88 (95% CI 1.31-2.68), implying that handwashing could reduce diarrhoea risk by 47%. When all studies, when only those of high quality, and when only those studies specifically mentioning soap were pooled, risk reduction ranged from 42-44%. The risks of severe intestinal infections and of shigellosis were associated with reductions of 48% and 59%, respectively. In the absence of adequate mortality studies, we extrapolate the potential number of diarrhoea deaths that could be averted by handwashing at about a million (1.1 million, lower estimate 0.5 million, upper estimate 1.4 million). Results may be affected by the poor quality of many of the studies and may be inflated by publication bias. On current evidence, washing hands with soap can reduce the risk of diarrhoeal diseases by 42-47% and interventions to promote handwashing might save a million lives. More and better-designed trials are needed to measure the impact of washing hands on diarrhoea and acute respiratory infections in developing countries. PMID- 12726976 TI - Recurrence in tuberculosis: relapse or reinfection? AB - The importance of reinfection as a cause for recurrence of tuberculosis is unclear and has potential public-health implications. We systematically searched published material for studies using DNA fingerprinting to provide data on the issue. Very few studies were designed for that particular research objective and/or report on a sufficient number of observations. Differences in methods--eg, case-definitions--seriously hamper comparisons between studies. The proportion of recurrences due to reinfection ranged between 0% and 100%; however, this figure cannot be a useful indicator since the two causes of recurrence--relapse and reinfection--are essentially independent. Only one study provides an estimate of the incidence of recurrence due to reinfection, indicating its importance for HIV infected patients in an environment with an unusually high tuberculosis incidence. We argue that apart from extreme situations like this one the problem of recurrence of tuberculosis due to reinfection has few implications for tuberculosis-control programmes. PMID- 12726977 TI - Haemophagocytic syndrome following Pseudomonas septicaemia. PMID- 12726978 TI - Clinical diagnosis of smear-negative pulmonary tuberculosis in low-income countries: the current evidence. AB - Sputum smear examination for acid-fast bacilli (AFB) can diagnose up to 50-60% of cases of pulmonary tuberculosis in well-equipped laboratories. In low-income countries, poor access to high-quality microscopy services contributes to even lower rates of AFB detection. Furthermore, in countries with high prevalence of both pulmonary tuberculosis and HIV infection, the detection rate is even lower owing to the paucibacillary nature of pulmonary tuberculosis in patients with HIV infection. In the absence of positive sputum smears for AFB, at primary care level, most cases of pulmonary tuberculosis are diagnosed on the basis of clinical and radiological indicators. This review aims to evaluate various criteria, algorithms, scoring systems, and clinical indicators used in low-income countries in the diagnosis of pulmonary tuberculosis in people with suspected tuberculosis but repeated negative sputum smears. Several algorithms and clinical scoring systems based on local epidemiology have been developed to predict smear negative tuberculosis. Few of these have been validated within the local context. However, in areas where smear-negative tuberculosis poses a major public-health problem, these algorithms may be useful to national tuberculosis programmes by providing a starting point for development their own context-specific diagnostic guidelines. PMID- 12726979 TI - Cutaneous histoplasmosis in a patient with systemic lupus erythematosis. PMID- 12726980 TI - Taking malaria transmission out of the bottle: implications of mosquito dispersal for vector-control interventions. AB - Most malaria transmission models assume enclosed systems of people, parasites, and vectors in which neither emigration nor immigration of mosquitoes is considered. This simplification has facilitated insightful analyses but has substantial limitations for evaluating control measures in the field. Here we show that mosquito dispersal can confound conventional approaches to evaluating malaria vector-control interventions, and explore this association with a model of two villages between which mosquito subpopulation exchange occurs. Exchange of mosquitoes between such subpopulations can readily explain the discrepancy between experimental efficacy measurements for insecticide-treated nets and their much lower apparent effectiveness when in use. Our results indicate that current approaches to assessing malaria interventions that confer community-level protection may be severely compromised by mosquito dispersal in many endemic settings. The true effectiveness of many vector-control methods may be much greater than previously appreciated and the application of such interventions should be consolidated into larger contiguous spatial units so that more effective local suppression of malaria can be achieved. Spatially explicit modelling formats that consider vector dispersal as a determinant of malaria transmission and control are needed urgently for rational planning and evaluation of efforts to roll back malaria. PMID- 12726981 TI - Scaling-up coverage with insecticide-treated nets against malaria in Africa: who should pay? AB - Insecticide-treated nets (ITNs) have been shown to reduce the burden of malaria in African villages by providing personal protection and, if coverage of a community is comprehensive, by reducing the infective mosquito population. We do not accept the view that scaling-up this method should be by making villagers pay for nets and insecticide, with subsidies limited so as not to discourage the private sector. We consider that ITNs should be viewed as a public good, like vaccines, and should be provided via the public sector with generous assistance from donors. Our experience is that teams distributing free ITNs, replacing them after about 4 years when they are torn and retreating them annually, have high productivity and provide more comprehensive and equitable coverage than has been reported for marketing systems. Very few of the free nets are misused or sold. The estimated cost would be an annual expenditure of about US$295 million to provide for all of rural tropical Africa where most of the world's malaria exists. This expenditure is affordable by the world community as a whole, but not by its poorest members. Recently, funding of this order of magnitude has been committed by donor agencies for malaria control. PMID- 12726983 TI - Joseph Malik Peiris--on the trail of pneumonia in Hong Kong interviewed by Pam Das. PMID- 12726987 TI - Seizing the opportunity. PMID- 12726988 TI - Inactivation of mprF affects vancomycin susceptibility in Staphylococcus aureus. AB - A chemically generated mutant of Staphylococcus aureus RN4220, GC6668, was isolated that had a fourfold increase in resistance to vancomycin. This phenotype reverted back to susceptibility by insertional mutagenesis with Tn917. In a selected set of revertants, Tn917 insertion was mapped to a unique chromosomal region upstream of mprF, a recently described gene that determines staphylococcal resistance to several host defense peptides. The genetic linkage between the vancomycin susceptibility and Tn917 insertion was then confirmed by transduction backcrosses into both GC6668 and GISA isolates, MER-S12 and HT2002 0127. Northern blot analysis, insertional inactivation and complementation experiments showed that mprF mediates vancomycin susceptibility in S. aureus. The inactivation of mprF by Tn917 insertion in HT2002 0127 caused a significant increase in the binding of vancomycin to the cell membranes. This observation serves as a likely mechanism of the increased vancomycin susceptibility associated with mprF inactivation. PMID- 12726989 TI - On the mammalian acetone metabolism: from chemistry to clinical implications. AB - Despite the description of the ways of acetone metabolism, its real role(s) is (are) still unknown in metabolic network. In this article, a trial is made to ascertain a comprehensive overview of acetone research extending discussion from chemistry to clinical implications. Mammals are quite similar regarding their acetone metabolism, even if species differences can also be observed. By reviewing experimental data, it seems that plasma concentration of acetone in different species is in the order of 10 microm range and the concentration dependent acetone metabolism is common to all mammals. At low concentrations of plasma acetone, the C3 pathways are operative, while at higher concentrations, the metabolism through acetate becomes dominant. Glucose formation from acetone may also contribute to the maintenance of a constant blood glucose level, but it seems to be only a minor source for that. From energetical point of view, an interorgan cooperation is suggested because transportable C3 fragments produced in the liver can serve as alternative sources of energy for the peripheral tissues in the short of circulating glucose. The degradation of acetoacetate to acetone contributes to the maintenance of pH buffering capacity, as well. Special attention is paid to the discussion of acetone production in diseases amongst which endogenous and exogenous acetonemiae have been defined. Acetonemiae of endogenous origin are due to the increased rate of acetone production followed by an increase of degrading capacity as cytochrome p450IIE1 (CYPIIE1) isozymes become induced. Exogenous acetonemiae usually resulted from intoxications caused by either acetone itself or other exogenous compounds (ethanol, isopropyl alcohol). It is highlighted that, on the one hand, isopropanol is also a normal constituent of metabolism and, on the other hand, the flat opinion that the elevation of its plasma level is a sign of alcoholism cannot further be held. The possible future directions of research upon acetone are depicted by emphasizing the need for the clear-cut identification of mammalian acetoacetate decarboxylase, and the investigation of race differences and genetic background of acetone metabolism. PMID- 12726990 TI - Heterogeneous inhibition of horseradish peroxidase activity by cadmium. AB - Inhibition of horseradish peroxidase (HRP) activity by cadmium was studied under steady-state kinetic conditions after preincubation of the enzyme with millimolar concentrations of Cd(2+) for various periods of time. The H(2)O(2)-mediated oxidation of o-dianisidine by HRP was used to assess the enzymatic activity. Cd(2+) was found to be either a noncompetitive inhibitor of HRP or a mixed inhibitor of HRP depending both on the duration of incubation with HRP and on Cd(2+) concentration. Furthermore, for the same inhibition type, K(i) values dropped as incubation time increased. These results suggested that Cd(2+) would slowly bind to the enzyme and progressively induce conformational changes. Spectrophotometric analysis showed that indeed Cd(2+) altered the heme Soret absorption band on binding HRP and exhibited a K(d) which decreased as the incubation time of HRP with Cd(2+) increased. Hill plots suggested a cooperative binding of up to three Cd(2+) ions per molecule of HRP. Thus, Cd(2+) binding to HRP resulted in progressive inhibition of enzymatic activity with a change in the inhibition type as the number of Cd(2+) ions per HRP molecule increased. Results also illustrated the potential danger of long-term exposure to heavy metals, even for enzymes with low affinity for them. PMID- 12726991 TI - Cathepsin B in osteoblasts. AB - Active cathepsin B has been found in cell extract and medium of human osteoblast like cells and MG-63 cells. The released form is stable at neutral and alkaline pH and, in both cell types, intracellular and extracellular cathepsin B activities are increased by interleukin-1 beta (IL-1beta) and parathyroid hormone (PTH). To evaluate the physiological role of cathepsin B in osteoblasts, we investigated the production and secretion of this enzyme in normal human synovial fibroblasts and modulation by IL-1beta and PTH. Lactate secretion concurrent with release of cathepsin B and comparable responses in osteoblasts were also examined. Our data show that synovial fibroblasts respond differently to treatment with the two agents, suggesting a cell-specific regulation of cathepsin B and possible involvement in osteoblast physiology. Cathepsin B involvement was then evaluated in the activation of plasminogen activator (PA) in MG-63 cells using two specific inhibitors of cathepsin B, CA074 and CA074-Me, in constitutive conditions and after treatment with IL-1beta. As results of PA activity obtained in the presence of IL-beta were in contrast with previous reports, we examined the activities of PA, pro-PA activated with trypsin, and plasmin in cell extract and media of MG-63 cells after 24-h treatment with IL-1beta. Results show that in normal conditions and in the presence of IL-1beta, cathepsin B is involved in the activation of PA. Moreover, IL-1beta stimulates PA, pro-PA activated by trypsin, and plasmin activity in medium, whereas in cell extract it stimulates pro-PA activated by trypsin and plasmin activity. IL-1beta has no effect on cell extract associated PA. PMID- 12726992 TI - Ozone impact on the photosynthetic apparatus and the protective role of polyamines. AB - One of the primary plant mechanisms protecting leaf cells against enhanced atmospheric ozone is the accumulation of polyamines, generally observed as an increase in putrescine level, and in particular its bound form to thylakoid membranes. Ozone-sensitive plants of tobacco (cultivar Bel W3) in contrast to ozone-tolerant Bel B, are not able to increase their endogenous thylakoid membrane-bound putrescine when they are exposed to an atmosphere with enhanced ozone concentration, resulting in reduction of their photosynthetic rates and consequently reduction in plant biomass formation. In comparison to the tolerant cultivar Bel B, a prolongation of ozone exposure thus can lead to typical visible symptoms (necrotic spots) in leaves of the sensitive plant. Exogenously manipulated increase of the cellular putrescine levels of the ozone-sensitive Bel W3 is sufficient to revert these effects, whereas a reduction in endogenous putrescine levels of the tolerant cultivar Bel B renders them sensitive to ozone treatment. The results of this work reveal a regulator role for polyamines in adaptation of the photosynthetic apparatus and consequently to its protection in an environment polluted by ozone. PMID- 12726993 TI - A trypsin inhibitor from Peltophorum dubium seeds active against pest proteases and its effect on the survival of Anagasta kuehniella (Lepidoptera: Pyralidae). AB - A novel trypsin inhibitor was purified from the seeds of Peltophorum dubium (Spreng.). SDS-PAGE under reducing conditions showed that the inhibitor consisted of a single polypeptide chain (ca. 20 kDa). The dissociation constants of 4 x 10( 10) and 1.6 x 10(-10) M were obtained with bovine and porcine trypsin, respectively. This constant was lower (2.6 x 10(-7) M) for chymotrypsin. The inhibitory activity was stable over a wide range of temperature and pH and in the presence of DTT. The N-terminal sequence of the P. dubium inhibitor showed a high degree of homology with other Kunitz-type inhibitors. When fed to the insect Anagasta kuehniella, in an artificial diet (inhibitor concentration 1.6%), the inhibitor produced approximately 56% and delayed the development of this lepidopteran. The concentration of inhibitor in the diet necessary to cause a 50% reduction in the weight (ED50) of fourth instar larvae was approximately 1%. The action of the P. dubium trypsin inhibitor (PDTI) on A. kuehniella may involve inhibition of the trypsin-like activity present in the larval midgut, resistance of the inhibitor to digestion by midgut enzymes and bovine trypsin, and association of the inhibitor with a chitin column and chitinous structures in the peritrophic membrane and/or midgut of the insect. PMID- 12726994 TI - Spectroscopic studies of the interaction of bichromophoric cyanine dyes with DNA. Effect of ionic strength. AB - Spectroscopic characteristics of a cyanine dye with two chromophores (biscyanine dye, BCD) in aqueous solutions and effects of NaCl and DNA upon these characteristics have been studied by optical absorption, circular dichroism (CD) and fluorescence spectroscopies. In homogeneous solutions, BCD is characterized by intense optical absorption (epsilon =1.33 x 10(5) M(-1) x cm(-1)) and weak fluorescence (phi(fl)=0.018) in the wavelength region greater than 600 nm. The dye forms H-aggregates at low concentrations (10(-6) M). NaCl stimulates the formation of both H- and J-aggregates of the dye at much lower dye concentrations, while DNA in low concentrations (<10(-6) M) stimulated the formation of just J-aggregates on the surface of the DNA molecule. Higher DNA concentrations induce the dye to disaggregate, and there exists an equilibrium between three dye forms: free monomers, J-aggregates and bound monomers, the maximum content of J-aggregates was observed at [DNA]/[BCD]=0.6+/-0.2 and total disaggregation at [DNA]/[BCD]=190+/-20. J-aggregates are characterized by phi(fl)=0.05 and bound monomers by phi(fl)=0.44. In the presence of NaCl, total disaggregation was observed at [DNA]/[BCD]=570+/-10 due to competition between Na(+) and the dye molecules for DNA electronegative binding sites. PMID- 12726995 TI - The role of N-linked glycosylation in determining the surface expression, G protein interaction and effector coupling of the alpha (alpha) isoform of the human thromboxane A(2) receptor. AB - In humans, thromboxane (TX) A(2) signals through two TXA(2) receptor (TP) isoforms, termed TPalpha and TPbeta, that diverge exclusively within the carboxyl terminal cytoplasmic domains. The amino terminal extracellular region of the TPs contains two highly conserved Asn (N)-linked glycosylation sites at Asn(4) and Asn(16). While it has been established that impairment of N-glycosylation of TPalpha significantly affects ligand binding/intracellular signalling, previous studies did not ascertain whether N-linked glycosylation was critical for ligand binding per se or whether it was required for the intracellular trafficking and the functional expression of TPalpha on the plasma membrane (PM). In the current study, we investigated the role of N-linked glycosylation in determining the functional expression of TPalpha, by assessment of its ligand binding, G protein coupling and intracellular signalling properties, correlating it with the level of antigenic TPalpha protein expressed on the PM and/or retained intracellularly. From our data, we conclude that N-glycosylation of either Asn(4) or Asn(16) is required and sufficient for expression of functionally active TPalpha on the PM while the fully non-glycosylated TPalpha(N4,N16-Q4,Q16) is almost completely retained within the endoplasmic reticulum (ER) and remains functionally inactive, failing to associate with its coupling G protein Galpha(q) and, in turn, failing to mediate phospholipase (PL) Cbeta activation. PMID- 12726996 TI - Purification and properties of two type-B alpha-L-arabinofuranosidases produced by Penicillium chrysogenum. AB - Two distinct extracellular alpha-L-arabinofuranosidases (AFases; EC 3.2.1.55) were purified from the culture filtrate of Penicillium chrysogenum 31B. The molecular masses of the enzymes were estimated to be 79 kDa (AFQ1) and 52 kDa (AFS1) by SDS-PAGE. Both enzymes had their highest activities at 50 degrees C and were stable up to 50 degrees C. Enzyme activities of AFQ1 and AFS1 were highest at pH 4.0 to 6.5 and pH 3.3 to 5.0, respectively. Addition of 10 mg/ml arabinose to the reaction mixture decreased the AFS1 activity but hardly affected AFQ1. Both enzymes displayed broad substrate specificities; they released arabinose from branched arabinan, debranched arabinan, arabinoxylan, arabinogalactan, and arabino-oligosaccharides. AFS1 also showed low activity towards p-nitrophenyl beta-D-xylopyranoside. An exo-arabinanase, which catalyzes the release of arabinobiose from linear arabinan at the nonreducing terminus, acted synergistically with both enzymes to produce L-arabinose from branched arabinan. PMID- 12726997 TI - DNA damage measured by liquid chromatography-mass spectrometry in mouse fibroblast cells exposed to oxidative stress. AB - Oxidative DNA damage can result from environmental factors, such as radiation, as well as from the untoward consequences of normal metabolic processes. It is of interest to assay oxidative DNA damage in cells and tissues because this damage has been implicated in human disease, particularly cancer. Eleven indicators of oxidative DNA damage have been measured by Liquid Chromatography-Mass Spectrometry (LC-MS) in DNA extracted from cells exposed to oxidative stress. Mouse fibroblast cells were exposed to hydrogen peroxide and to UVC light and to the combined action of both agents. Significant increases of the 8-oxo-7,8 dihydropurine lesions over background were detected. Significant increases of the formamido lesions resulting from breakdown of pyrimidine bases were also observed. Of special interest was the observation of double lesions, tandem combinations of both aforementioned lesions, in cells exposed to oxidative stress. PMID- 12726998 TI - Induction of hsp70 in transgenic Drosophila: biomarker of exposure against phthalimide group of chemicals. AB - The expression of stress genes is suggested to be a potentially sensitive indicator of any chemical or physical assault. This led us to explore the possibility of using expression of one of the major stress genes, hsp70, in Drosophila as a biomarker against phthalimide group of chemicals, which may accordingly provide an early indication of exposure to these hazardous chemicals. We exposed third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ) Bg(9) to different concentrations of the test chemicals (Captan, Captafol and Folpet) for various time intervals (2-48 h) to evaluate expression of hsp70 by X gal staining, ONPG assay and whole organ in situ immunohistochemistry. The study was further extended to examine the effect of the said chemicals on development of the organism and tissue damage occurring in them, thus raising the possibility of evaluating comparative deleterious effect inducing potential of the test chemicals. Our results showed a strong hsp70 expression in the Captafol-exposed larvae followed by weaker expression in Captan- and Folpet-treated larvae. The effect was further reflected on development as revealed by a delay in emergence of the flies by 3 days in 200 ppm Captafol-exposed group. Hsp70 was found not to be induced at 0.0002 ppm Captafol and at 0.002 ppm Captan and Folpet. The present study suggests that (a). hsp70 induction is sensitive enough to be used as a biomarker against phthalimide group of chemicals, (b). amongst the three test chemicals, Captafol is the most deleterious compound followed by Captan and Folpet, (c). 0.0002 ppm for Captafol and 0.002 ppm for Captan and Folpet, respectively, can be regarded as no observed adverse effect level (NOAEL). PMID- 12726999 TI - Binding and protection of porphyrins by glutathione S-transferases of Zea mays L. AB - Glutathione S-transferases (GSTs) are multi-functional enzymes, known to conjugate xenobiotics and degrade peroxides. Herein, we report on the potential of four Zea mays GST isoforms (Zm GST I-I, Zm GST I-II, Zm GST II-II and Zm GST III-III) to act as binding and protection proteins. These isoforms bind protoporphyrin IX (PPIX), mesoporphyrin, coproporphyrin, uroporphyrin and Mg protoporpyhrin, but do not form a glutathione conjugate. The binding is non covalent and inhibits GSTs enzymatic activity, dependent on the type of the porphyrin and GST isoform tested. I(50) values are in the range of 1 to 10 microM for PPIX, the inhibition by mesoporphyrin and Mg-protoporphyrin (Mg-PPIX) is two to five times less. The mode of binding is non-competitive for the hydrophobic substrate and competitive for glutathione. Binding affinities (K(D) values) of the GST isoforms are between 0.3 and 0.8 microM for coproporphyrin and about 2 microM for mesoporphyrin.Zm GST III-III prevents the nonenzymatic autoxidation of protoporphyrinogen to the phytotoxic PPIX. Zm GST II-II can reduce the oxidative degradation of hemin. This points to a specific ligand role of distinct GST isoforms to protect tetrapyrroles in the plant cell. PMID- 12727000 TI - The Jing-Mai connections of the heart. AB - BACKGROUND: The Jing-Mai (variously translated as the Channel, Vessel or Meridians), as described by traditional Chinese medicine, probably exists and has represented the connections between various parts of human body during embryonic development. According to the Chinese theories, there are 14 major Jing-Mai within the human body, of which four are directly connected with the Heart. METHODS: The described paths of the four Jing-Mai were compared with features of congenital syndromes involving particular types of congenital heart defects. RESULTS: Specific correlation seem to exist between such four Jing-Mai and known developmental mechanisms underlying various congenital heart defects: the Kidney Jing-Mai-ectomesenchymal tissue migration abnormalities; the Spleen Jing-Mai situs and looping defects; the Heart Jing-Mai-abnormal cell death; the Small Intestine Jing-Mai (and the Heart Jing-Mai)-extracellular matrix anomalies. CONCLUSIONS: The Chinese theories seem to provide some intriguing insights into the pathogeneses of congenital heart defects. The Jing-Mai seems to distinguish from, but nevertheless have a close relationship with the blood vessels. Utilization of the Jing-Mai will probably enable a better understanding and development of new treatments for cardiovascular diseases. PMID- 12727001 TI - Familial hypercholesterolemia--improving treatment and meeting guidelines. AB - Familial hypercholesterolemia (FH) is a common, inherited disorder that affects around one in 500 individuals in the heterozygous form. By the year 2001, more people in the US had FH than were infected by the human immunodeficiency virus. The disease is caused by mutations within the low-density lipoprotein (LDL) receptor gene. FH is associated with elevated plasma LDL-cholesterol (LDL-C) levels, xanthomatosis, early onset of atherosclerosis and premature cardiac death. Patients with heterozygous FH commonly have plasma LDL-C levels that are two-fold higher than normal, while homozygotes have four- to five-fold elevations in plasma LDL-C. Although FH patients have a high risk of developing premature coronary heart disease (CHD), they remain underdiagnosed and undertreated. Early detection of FH is critical to prolonging the life of these patients. Once identified, patients with heterozygous FH can be placed on a diet and drug management program. As the most efficacious and well-tolerated agents, hydroxy methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) are usually the drugs of first choice; bile acid sequestrants, niacin, and occasionally fibrates may be used as supplemental agents. Statins may also provide a realistic option for the treatment of some FH homozygotes with genes that produce partially functional LDL receptors. However, a number of patients are still failing to reach treatment guidelines even with the most effective of the currently available statins. The development of new more efficacious statins or the use of new combination therapies such as statins with the cholesterol absorption inhibitor, ezetimibe may help to reduce the current problem of undertreatment in FH patients. PMID- 12727002 TI - Echocardiographic changes and predictors of arrhythmia recurrence after long-term use of the atrial defibrillator. AB - BACKGROUND: The patient-activated atrial defibrillator allows patients to cardiovert themselves from atrial fibrillation soon after the onset of symptoms. The long-term effects of early cardioversion from persistent atrial fibrillation on left ventricular performance and left atrial size are unknown. METHODS: Eighteen patients, mean age 63.4, 83% male, had the Jewel((R)) AF atrial defibrillator implanted for persistent atrial fibrillation only. Transthoracic echocardiography was performed 3-monthly following implant. Parasternal long axis measurements were taken using conventional M-mode techniques. RESULTS: Over follow-up of 28.0+/-9 months, 377 episodes of persistent atrial fibrillation were terminated by patient-activated cardioversion (median 15 per patient). Echocardiographic measurements at implant were; left atrium 44+/-6 mm, left ventricular end-diastolic diameter 49+/-7 mm, left ventricular end-systolic diameter 34+/-7 mm, fractional shortening 33+/-10% and ejection fraction 65+/ 17%. After 1 year there had been a significant decrease in mean left atrial size to 41+/-6 mm (P=0.02) and an increase in mean ejection fraction to 73+/-8% (P=0.04). At long-term follow-up however, all parameters reverted to pre-implant levels. Baseline echocardiographic variables did not predict which patients would demonstrate serial increases in sinus rhythm duration between shocks during long term follow-up. Patients on antiarrhythmic drug therapy however were more likely to demonstrate "sinus rhythm begetting sinus rhythm". CONCLUSIONS: Use of the atrial defibrillator for spontaneous persistent atrial fibrillation is associated with a medium-term (1 year) reduction in left atrial size and an increase in ejection fraction. These changes were not maintained in the long-term. Synergistic therapy with antiarrhythmic drugs may prolong periods of sinus rhythm between arrhythmia recurrences. PMID- 12727003 TI - Myocardial time intervals preceding left ventricular filling in chronic coronary artery disease: value of a decreased septal ejection time. AB - The aim was to assess the capabilities of a two-segment myocardial recording to recognize patients with an underlying chronic ischemic process as a fast screening from controls, prior to the usual segment-to-segment tissue Doppler echocardiographic assessment of ischemia. Ischemia generates systolic and relaxation abnormalities. A flow Doppler index of global systolic and diastolic myocardial performance was recently drawn from time durations studied by coupling isovolumic relaxation (IR) to preejection (PEP)/ejection (ET) ratio (PEP/ET). We derived a similar tissue Doppler approach to the period preceding the left ventricular filling: PEP', the ejectional inward wall motion representing ET' and the prefilling (PreFg) period ranging from the end of ET' to the onset of the outward wall motion approximating IR, were measured and ratios calculated between variables. Spectral tissue Doppler was applied to septal and posterior walls of 28 patients with proven chronic coronary artery disease and preserved left ventricular function and of 12 age-matched controls. Data were compared with global flow data. Global information did not differentiate both groups, save for IR (sensitivity 32%, specificity 57%). In patients, tissue Doppler mean values of single variables (P=0.004-0.0006) and ratios (P=0.03-0.002) significantly differed from controls. Moreover, septal ET' differentiated 13 patients with one vessel (219+/-34 ms) from 10 with two-vessel disease (158+/-70 ms, P=0.01). Sensitivity and specificity of a septal ET'<190 ms for a two-vessel disease were 80%. The two-segment tissue Doppler echocardiographic study provided a rapid screening of patients versus controls and helped to predict the number of diseased vessels. PMID- 12727004 TI - Natriuretic peptide and echocardiography after operation of atrial septal defect. AB - Patients benefit from surgical seclusion of atrial septal defect but have excessive cardiovascular morbidity after the operation. We evaluated haemodynamics and looked for abnormalities of cardiac structures and function late after surgical seclusion of the defect. Serum N-terminal natriuretic peptide measurement and transthoracic and transoesophageal echocardiography were performed in 61 patients aged 43+/-15 years (mean+/-standard deviation) 21+/-5 years after surgery. The findings were compared with 67 control subjects. The patients had higher serum N-terminal atrial natriuretic peptide concentration than the control subjects (0.40+/-0.32 vs. 0.24+/-0.12 nmol/l, P=0.0001). Peptide levels correlated with current age (P=0.0001) and age at operation (P=0.0014), but not with age in the control subjects. In the patients, echocardiography measurements of cardiac dimensions correlated with hormone levels (atrial natriuretic peptide concentration with left atrial end-systolic diameter (P=0.042), left ventricular end-diastolic (P=0.021) and end-systolic diameter (P=0.042). There were only 10 patients (16%) without any abnormality in echocardiography. Their peptide concentration was 0.25+/-0.18 nmol/l (P=not significant compared to the control subjects). The association between increasing N-terminal atrial peptide levels and operation age together with echocardiography findings support the clinical consensus of treating atrial septal defect patients in their childhood and adolescence. PMID- 12727005 TI - Differential mononuclear cell activity and endothelial inflammation in coronary artery disease and cardiac syndrome X. AB - BACKGROUND: Circulating mononuclear cells could be activated with endothelial inflammation in patients with coronary artery disease (CAD). In some patients with normal coronary angiograms, myocardial ischemia could also present with coronary microvascular dysfunction (cardiac syndrome X). This study was undertaken to investigate whether mononuclear cell activation and endothelial inflammation can present in syndrome X patients. METHODS: We evaluated the biochemical parameters, circulating soluble adhesion molecules, circulating superoxide free radicals, and mononuclear cell activity in 32 patients with syndrome X, 34 with angiographically documented CAD, and 17 age- and gender matched healthy control subjects. RESULTS: Compared to that in control subjects, plasma high-density lipoprotein was reduced (P<0.001) and insulin to glucose ratio increased (P=0.02) in CAD patients. Circulating level of soluble intracellular adhesion molecule-1 was significantly higher in both syndrome X and CAD patients than in control subjects (P<0.01), whereas the levels of soluble vascular cell adhesion molecule (P=0.02) and von Willebrand factor (P=0.01) were increased in CAD patients only. The peak (P<0.001) and total counts of superoxide free radicals in whole blood (P<0.001) was significantly higher in syndrome X patients than in the other two groups. However, phorbol-12-myristate-13-acetate induced superoxide free radical generation of mononuclear cells was increased in CAD (10.5+/-4.6%, P=0.01) but not in syndrome X patients (8.7+/-2.0%) as compared with control subjects (7.7+/-0.5%). CONCLUSIONS: The results indicated that the activity of mononuclear cells was increased with significant endothelial inflammation and injury in CAD patients. In syndrome X patients, though circulating superoxide free radicals were increased, there was minimal endothelial inflammation without mononuclear cell activation. The relatively preserved lipid and metabolic profiles might contribute to less vascular inflammation in syndrome X patients. PMID- 12727006 TI - Duration of symptoms in patients with infective endocarditis. AB - Despite progress in the management of infective endocarditis, delays in diagnosis or prior antimicrobial treatment may adversely influence the symptom duration and outcome. The duration of symptoms in patients with infective endocarditis was studied in 683 cases among 653 patients with 703 episodes of the disease; patients were hospitalized within 10 days of symptom onset in 169 (24.7%) cases. Antimicrobial therapy before hospital admission was administered to 257 (36.5%) patients. Overall mortality was 25.6%. Symptom duration was longer when antimicrobials were administered before diagnosis (58.8+/-78.1 vs. 44.8+/-54.9 days), when vegetations were detected on echocardiogram (53.5+/-68.2 vs. 38.8+/ 47.3) and among patients admitted before 1990 (42.3+/-67.1 vs. 54.2+/-62.4 days). Symptom duration was shorter in patients with prosthetic valve endocarditis (26.8+/-34.2 vs. 59.3+/-71.6 days). In 54 (26.5%) episodes of prosthetic valve endocarditis, patients had symptoms for more than 30 days. Staphylococcus aureus was the most frequent agent among patients with symptoms up to 10 days (41.2%) and Streptococcus among those with symptoms over 20 days (53.9%). Symptom duration did not significantly differ in regard to medical (51.3+/-69.2 days) or surgical (46.7+/-55.7 days) treatment. Mortality increased as symptom duration decreased and was highest for patients who experienced symptoms for less than 10 days (36.1%). In some patients medical care may be delivered relatively late in the course of infective endocarditis. Administration of antibiotics previous to hospital admission increased duration of symptoms, and cardiac valve prosthesis, staphylococcal infection and death were associated with more acute disease. PMID- 12727007 TI - Contrasting roles of a novel K+ channel blocker and a K+ channel opener on electro-mechanical activity in canine heart tissue. AB - We tested the effects of a potassium channel opener diazoxide on the action potential duration (APD) and contractile force changes in canine Purkinje tissue induced by a novel class III anti-arrhythmic agent (C3A), KCB-328 (0.5 microM) with 3,4-dimethoxyphenethyl ring structure (0.5 microM). KCB-328 shortened APD(25) by 8.3+/-2.1%, prolonged APD(50) and APD(90) by 31.2+/-5.3 and 50.0+/ 7.1%, respectively. Diazoxide (0.1 mM) shortened APD at all levels by 58.3+/-8.1, 54.1+/-6.1, and 42.8+/-5.8%, respectively. In the presence of diazoxide, KCB-328 still prolonged APD(50) and APD(90) (12.5+/-3.8 and 26.8+/-5.9%, respectively). KCB-328 increased force of contraction in a dose-dependent manner. KCB-328 increased force less in the presence of diazoxide. Administration of diazoxide only, reduced force of contraction. We conclude that APD prolongation by KCB-328 may occur even in the presence of diazoxide. It is not sufficient for the restoration of already diminished contractile force and that such an APD prolongation may be unrelated to the restoration of force of contraction even though both are most often seen to occur simultaneously. PMID- 12727008 TI - Electrocardiographic algorithm for assignment of occluded vessel in acute myocardial infarction. AB - BACKGROUND: This study was performed to elaborate an electrocardiographic (ECG) algorithm enabling assignment of an occluded coronary artery in acute myocardial infarction (AMI). PATIENTS AND INTERVENTIONS: In 109 patients (age, 59+/-12 years) with AMI (pain onset, 3.6+/-1.7 h), coronary angiography with PTCA/stenting of the culprit lesion was performed. The diagnosis of AMI was confirmed by emergency coronary angiography and laboratory analyses. Admission ECG parameters (amplitude of R-wave, ST-segment deviation, presence of Q-wave, deflection of T-wave) in standard 12-lead ECG plus extended (V(3)R to V(6)R and V(7-9)) leads were subjected to classification and regression tree (CART) analysis. RESULTS: Continuous CART analysis assessed ST-segment deviations in V(2) and V(5)R. AMI of the left anterior descending (LAD), right coronary artery (RCA) and left circumflex coronary artery (CX) were correctly classified in 94, 64, and 91% of cases, respectively. Dichotomised CART analysis assessed ST segment deviations in V(2), V(5)R, and aVF. True classification rates for LAD, RCA, and CX amounted to 84, 74, and 71%, respectively. CONCLUSIONS: Dichotomised CART analysis is a simple means of differentiation of CX from RCA occlusion during AMI. PMID- 12727010 TI - Ablation of atrial reentrant tachycardia in children with silent atrium. PMID- 12727009 TI - Dissolution of a huge spontaneous coronary artery thrombus with a new antiplatelet agent and coronary angioplasty. AB - A huge thrombus was observed in the proximal portion of the right coronary artery (RCA) in a 75-year-old patient. He presented with symptoms of cardiogenic shock due to acute myocardial infarction. Glycoprotein IIb/IIIa inhibitor (Tirofiban) was administered intra-coronary first, coronary angioplasty was performed later and, dramatic thrombus dissolution was observed on control coronary angiography. PMID- 12727011 TI - Risk of recurrent thrombotic events in patients with coronary heart disease and left ventricular dysfunction. PMID- 12727013 TI - Acute thrombosis of a separate major coronary artery during initially successful thrombolytic therapy. PMID- 12727012 TI - An unusual cause of enhancement of the swirling motion of left atrial spontaneous echo contrast. PMID- 12727014 TI - Sildenafil in secondary pulmonary hypertension. PMID- 12727015 TI - Decreased nocturnal synthesis of melatonin in patients with coronary artery disease. AB - In human beings, cardiovascular activity has a distinct circadian variation: Heart rate, blood pressure, and vascular tone decrease at night. Nocturnal cardiovascular blunting is at least partially linked to the autonomic activity and increased risk of cardiac and cerebral events. To assess whether decreased nocturnal melatonin synthesis and secretion in coronary artery disease (CAD), we investigated nocturnal secretion pattern of melatonin in patients with CAD and healthy subjects. The present study performed in 16 patients with angiographically documented CAD (aged 46-71 years) and in nine healthy controls (aged 36-66 years). Blood samples were collected every 2 h between 22:00 and 08:00 h. Melatonin levels were measured with a commercially available radioimmunoassay kit. We found large interindividual variation in the pattern of melatonin secretion in both groups. Patients with CAD secreted less nocturnal melatonin at 02:00, 04:00 and 08:00 h than control subjects (P=0.014, P=0.04 and P=0.025, respectively). Peak and Delta melatonin (peak-lowest melatonin) were found lower in patients with CAD (48.6 [19.1-75.4] vs. 131.4 [67.8-137.2] pg/ml, P=0.006 and 43 [10.5-68.5] vs. 107.6 [55.7-113.1] pg/ml, P=0.002, respectively). Peak time of melatonin secretion was observed earlier in patients with CAD (02:00 h [23:00-02:00 h] vs. 03:45 h [02:00-05:00 h], P=0.04). Our study provides useful and preliminary information about decreased nocturnal melatonin synthesis and release in patients with CAD might help physicians in managing these patients. PMID- 12727016 TI - Complete endocardial cushion defect (complete atrioventricular canal) manifested in adult life by Streptococcus mitis endocarditis of the common atrioventricular valve. PMID- 12727017 TI - Malignant ventricular arrhythmias during coronary angiography. PMID- 12727018 TI - Management of a patient with active rheumatoid arthritis and suspected tuberculosis causing effusive-constrictive pericarditis. AB - In the following case report we present a patient who has been admitted for pericardial effusion causing cardiac compression with active rheumatoid arthritis and suspected tuberculosis. The patient was successfully treated with intravenous pulse steroid for active rheumatoid arthritis, with prophylactic anti tuberculosis agents for suspected tuberculosis and with surgical pericardiectomy for the thickened pericardium as well as recurrent pericardial effusion. PMID- 12727019 TI - Balloon dilatation of isolated congenital tricuspid stenosis. PMID- 12727020 TI - Epidermal basement membrane zone components: ultrastructural distribution and molecular interactions. AB - The epidermal basement membrane zone (BMZ) comprises a multiprotein complex that aids the attachment of epidermal keratinocytes to the underlying dermis. In the last 5 years, our understanding of epidermal BMZ morphology, structure and function has dramatically improved. A complex network of molecular interactions has recently been identified that strengthen dermal-epidermal adhesion. Special attention will be paid to the recently identified network of interactions between BMZ components, including alpha6beta4 integrin interactions, the extracellular hook domain of collagen XVII and the looping structure of collagen VII molecules. We summarize some of the recent advances in the understanding of the biology and interactions of BMZ components from an ultrastructural and molecular perspective. PMID- 12727021 TI - Human keratinocytes express fractalkine/CX3CL1. AB - BACKGROUND: fractalkine/CX3CL1 is a unique chemokine that has properties of both chemoattractants and adhesion molecules. The major source of this chemokine in the skin is still controversial. OBJECTIVE: studies were undertaken to determine the expression of fractalkine in human skin. METHODS: RT-PCR, Western blotting, and immunostaining were performed with normal human epidermal keratinocytes (NHEK) and HaCaT cells, human keratinocyte cell line, for the presence of fractalkine. Biopsy specimens of normal and diseased skin were also investigated. RESULTS: we identified that NHEK and HaCaT cells expressed fractalkine mRNA and protein. The combination of tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma upregulated their expression by NHEK. Immunohistochemistry demonstrated fractalkine expression in keratinocytes in lichen planus and psoriasis vulgaris. RT-PCR also showed that lesional skin of psoriatic patients expressed higher levels of fractalkine mRNA than non-lesional skin from the same patients. CONCLUSION: these results suggests that keratinocytes strongly express fractalkine in lichen planus and psoriasis vulgaris and that the fractalkine CXC3CR1 system in the diseased skin can be a target for the treatment. PMID- 12727022 TI - A novel P gene missense mutation in a Japanese patient with oculocutaneous albinism type II (OCA2). AB - BACKGROUND: Oculocutaneous albinism type II (OCA2) is an autosomal recessively inherited disorder, characterized by white hair and skin, and loss of pigment in the eyes. Mutations in P gene have been shown to result in OCA2. So far, two cases have been reported from Japan. OBJECTIVE: We had an opportunity to examine a case of albinism, and screened the mutations of tyrosinase and P gene. METHODS: Genomic DNA was prepared from peripheral leukocytes. All of the exons and flanking introns of tyrosinase and P gene were PCR-direct-sequenced. RESULTS: Although no mutations were found in tyrosinase, we found two missense substitutions, A481T and Q799H in P gene. The A481T has previously been shown to result in partial function of the P protein. CONCLUSION: The Q799H mutation is not a common polymorphism among normal Japanese, seems most likely to be a pathological OCA2 mutation among Japanese with this form of albinism. PMID- 12727023 TI - Downregulation of NF-kappaB activation in human keratinocytes by melanogenic inhibitors. AB - BACKGROUND: Exposure of skin cells, particularly keratinocytes to various nuclear factor-kappaB (NF-kappaB) activators (e.g. tumor necrosis factor-alpha, interleukin-1, lipopolysaccharides, and ultraviolet light) leads to phosphorylation and degradation of the inhibitory protein, IkappaB. Liberated NF kappaB is translocated into the nucleus where it can change or alter expression of target genes, resulting in the secretion of extracellular signaling molecules including melanotrophic factors affecting melanocyte. OBJECTIVE: In order to demonstrate the possible role of NF-kappaB activation on the synthesis of melanotrophic factors from the keratinocytes, the activities of NF-kappaB induced by melanogenic inhibitors (MIs) were determined in human HaCaT keratinocytes transfected with pNF-kappaB-SEAP-NPT plasmid. Transfectant cells released the secretory alkaline phosphatase (SEAP) as a transcription reporter in response to the NF-kappaB activity and contain the neomycin phosphotransferase (NPT) gene for the dominant selection marker for geneticin resistance. METHODS: MIs such as niacinamide, kojic acid, hydroquinone, resorcinol, arbutin, and glycolic acid were preincubated with transfectant HaCaT cells for 3 h and then ultraviolet B (UVB) was irradiated. NF-kappaB activation was measured with the SEAP reporter gene assay using a fluorescence detection method. RESULTS: Of the MIs tested, kojic acid (IC(50)=60 microM) was found to be the most potent inhibitor of UVB upregulating NF-kappaB activation in transfectant HaCaT cells, which is followed by niacinamide (IC(50)=540 microM). Pretreatment of the transfectant HaCaT cells with the MIs, especially kojic acid and niacinamide, effectively lowered NF kappaB binding measured by electrophoretic mobility shift assay. Furthermore, these two inhibitors remarkably reduced the secretion level of IL-6, one of melanotrophic factors, triggered by UV-radiation of the HaCaT cells. CONCLUSION: These observations suggest that MIs working at the in vivo level might act partially through the modulation of the synthesis of melanotrophic factors in keratinocyte. PMID- 12727024 TI - Predominant cell-mediated immunity in the oral mucosa: gene gun-based vaccination against infectious diseases. AB - BACKGROUND: Direct immunization via epithelial surfaces has been considered for many vaccine approaches, including DNA vaccines. It remains to be determined, however, which body site is suitable for genetic vaccination. OBJECTIVE: To characterize the effects of the oral mucosa-mediated genetic vaccination, we compared antigen-specific immune responses of the oral mucosal DNA vaccine to the flank skin vaccination against influenza virus and malaria parasite. METHODS: DNA vaccines against the influenza A/WSN/33 (H1N1) hemagglutinin and the malaria Plasmodium berghei circumsporozoite protein were administered respectively three times at 3-week intervals into the oral mucosa, skin, or liver of hamsters. The effects of their vaccine were evaluated by antigen-specific antibody production and cell-mediated killing activity. Furthermore, the in vivo malaria challenge test was also performed after the vaccination. RESULTS: Significant specific antibody production was not observed in each case, but interferon-gamma production and cell-mediated killing activity were strongly induced in splenic lymphocytes from hamsters with the oral vaccination. The in vivo malaria challenge after the oral mucosal vaccination significantly delayed the blood appearance day of the parasites in comparison with other immunization sites (P<0.05). CONCLUSION: These results suggest that gene immunization via the oral mucosa may induce cell-mediated immunity more efficiently than via the skin or liver, and that the oral mucosa may be one of the most suitable tissues for gene gun-based DNA vaccination against infectious diseases. PMID- 12727025 TI - Effects of high glucose on NO synthesis in human keratinocyte cell line (HaCaT). AB - BACKGROUND: There is a possibility that alteration of nitric oxide (NO) synthesis by high glucose leads to a variety of diabetic complications. OBJECTIVE: In this study, we examined whether NO synthesis is altered by high glucose in spontaneously immortalized human keratinocyte cell line (HaCaT) that have three isoforms of NO synthases (NOS). METHODS: We measured NO end product nitrite in the culture medium using the Griess reagent and analyzed mRNA expression of three isoforms of NOS in HaCaT cells by RT-PCR. RESULTS: High glucose enhanced constitutively produced NO production in HaCaT cells, which persisted for 10 days and was attenuated by an inhibitor of protein kinase C (PKC), without altering eNOS/nNOS mRNA levels. Cytokine stimulation induced iNOS mRNA in HaCaT cells. Pretreatment with high glucose for 24 h enhanced cytokine-induced NO production in HaCaT cells. However, when these cells were exposed to high glucose for 10 days, cytokine treatment did not induce iNOS mRNA and nitrite production. CONCLUSION: These diverse alterations in NO production by high glucose may be involved in impaired host-defense and wound healing in the skin of diabetic patients. PMID- 12727026 TI - CD56: a useful marker for diagnosing Merkel cell carcinoma. AB - BACKGROUND: Merkel cell carcinoma (MCC) of the skin is an aggressive but rare malignant neuroendocrine tumor. For its pathological diagnosis, we use a panel of immunohistochemical markers, such as cytokeratin 20 (CK 20), epithelial membrane antigen (EMA), chromogranin A, neuron specific enolase (NSE), synaptophysin, and Leu7 (CD57) to demonstrate its epithelial and neuroendocrine features. CD56, or neural cell adhesion molecule (NCAM), has been demonstrated recently as the tumor marker of the pulmonary neuroendocrine cell system. Its expression in MCC, however, has still rarely been investigated. Furthermore, in such very few previous studies on NCAM expression in MCC, all the tumor cells were not necessarily demonstrated to express NCAM. OBJECTIVES: To study the immunoreactivity of CD56 in MCC, especially using a monoclonal antibody of a clone 1B6, different from those adopted in the previous reports. METHODS: We reexamined CD56 expression immunohistochemically in five MCC cases, along with the conventional panel of markers described above, using paraffin-embedded tissue sections. RESULTS: CD56 revealed the most diffuse and intense positive staining, which was noted along the cell borders, in all specimens compared with other neuroendocrine tumor markers. CONCLUSIONS: The results of our study indicate that CD56, especially a new monoclonal antibody (clone 1B6), is a useful immunohistochemical marker for MCC. PMID- 12727027 TI - Upregulated expression of human beta defensin-1 and -3 mRNA during differentiation of keratinocyte immortalized cell lines, HaCaT and PHK16-0b. AB - BACKGROUND: Human beta-defensins (hBDs) belong to a group of antimicrobial peptide that are expressed in the epithelial cells. OBJECTIVE: The present study investigated mRNA expression levels of the beta-defensins, hBD-1, -2 and -3, in human keratinocytes during differentiation in vitro. METHODS: Immortalized keratinocyte cell lines, HaCaT and PHK16-0b, were used in this study; in order to stimulate differentiation, the Ca(2+) concentration in the growth media was increased from 0.3 to 1.8 mM. RESULTS: Four days after the increase, the expression levels of hBD-1 and -3 were increased in both cell lines, followed by an increase in the mRNA levels of the differentiation markers, involucrin and keratin 10. No increased expression of hBD-2 was observed. CONCLUSION: The results indicate that keratinocyte differentiation may stimulate hBD-1 and -3 expression in stratified squamous epithelia. PMID- 12727028 TI - Paradoxical effect of trichloroacetic acid (TCA) on ultraviolet B-induced skin tumor formation. PMID- 12727029 TI - Candida albicans induced interleukin 8 production by human keratinocytes. PMID- 12727031 TI - Clinically important changes-what's important and whose change is it anyway? PMID- 12727032 TI - High dose opioids in pediatric palliative care. PMID- 12727034 TI - Other benefits of an opioid "agreement". PMID- 12727033 TI - A retrospective comparison of the dose ratio between subcutaneous and oral ketamine. PMID- 12727036 TI - Chronic testicular pain in a patient with diverticulitis. PMID- 12727037 TI - Clinically important changes in acute pain outcome measures: a validation study. AB - The purpose of this study was to validate the changes in acute pain measurement scales that are most strongly associated with a patient-determined indicator of clinical importance. Measures of pain intensity and pain relief are commonly used outcomes in therapeutic clinical trials. Recent studies of the properties of acute pain measures have provided data defining the cut-off points that are best associated with clinically important differences. Validation of these findings in another clinical trial data set is important. Data were obtained from the titration phase of a recently conducted randomized controlled clinical trial of oral transmucosal fentanyl citrate (OTFC), which compared OTFC to immediate release morphine sulfate (MSIR) for the treatment of cancer-related acute breakthrough pain. Changes in pain intensity and pain relief were recorded every 15 minutes for 60 minutes and global medication performance recorded at the end of each study pain episode. At any titration step, if the patient felt that the first dose of the study medication did not provide adequate relief within 30 minutes, an additional rescue medication could be taken. To find the level of each pain scale best associated with this measure of the adequacy of pain relief, the calculated sensitivity, specificity, and accuracy for different cut-off points of the measured pain scales were compared to whether or not the patient needed rescue medication. The overall ability of the pain measures to discriminate episodes for which a rescue was not needed was calculated using area under the receiver operating characteristics (ROC) curves. Data were analyzed from 134 OTFC-naive patients who collected data on 1307 episodes of breakthrough pain. Using the criteria of a balanced sensitivity and specificity, the best cut off points were determined to be: 33% for the percent pain intensity difference; > or =2 for the raw pain intensity difference on a 0-10 numeric rating scale; > or =2 (i.e., moderate or better) for pain relief; > or =33% for the percent maximum total pain relief; and > or =2 (good or better) for global medication performance. ROC area under the curve ranged from 0.839 to 0.862 for each of the pain measures listed above, calculated at 60 minutes. These data indicate that the pain scale cut-off points that are best associated with a patient-derived measure of a clinically important difference closely approximate those found in an earlier study. ROC analysis provided evidence that the overall pain measures were strongly associated with not requiring an "additional dose of rescue medication." Thus, the cut-off points determined for these pain scales provide a good surrogate measure of a patient-determined clinically important response. This provides support for the usefulness of these values in future clinical trials of pain therapy. PMID- 12727038 TI - Breast cancer patient perception of the helpfulness of a prompt sheet versus a general information sheet during outpatient consultation: a randomized, controlled trial. AB - The purpose of this study was to determine the helpfulness of a prompt sheet versus a general information sheet for patient communication with physicians. Sixty women with breast cancer attending their first outpatient consultation with a breast medical oncologist were randomized to receive either a prompt sheet (PS) or a general information (GI) sheet regarding breast cancer. Analysis of the results found that helpfulness of the written material was rated higher in the PS group (8.5 +/- 2) than the GI group (6.2 +/- 3.6), P = 0.005. The mean score of helpfulness in communicating with physicians was 7.9 +/- 2.4 and 5.7 +/- 3.8, respectively, P = 0.01. There were no significant differences between the groups in the average total number of questions asked by the patients or average physician or patient speaking time. We conclude that a disease-specific prompt sheet provided before medical encounters may assist in communication between patients and physicians. PMID- 12727039 TI - Measuring the symptom experience of seriously ill cancer and noncancer hospitalized patients near the end of life with the memorial symptom assessment scale. AB - The objectives of this study were twofold: (1) to explore and compare the symptom experience of seriously ill hospitalized cancer and noncancer patients near the end of life using the Memorial Symptom Assessment Scale (MSAS) and (2) to determine if the MSAS is a valid and useful measure of symptom distress for patients with noncancer conditions. This was a prospective cohort study of hospitalized patients with end-stage congestive heart disease, chronic pulmonary disease, cirrhosis, or metastatic cancer. Eligible patients were interviewed to ascertain symptom prevalence, severity and distress using the MSAS and levels of fatigue using the Piper Fatigue Scale (PFS). Sixty-six patients with metastatic cancer and 69 patients with end-stage disease were enrolled in the study. There was a significant difference in the prevalence of selected physical symptoms, but not psychological symptoms, between cancer and noncancer patients. There were no significant differences in symptom distress scores, a computed score of frequency, severity and distress, if the symptom was present. In both groups the principal components factor analysis with varimax rotation yielded one factor comprising psychological symptoms and a second factor comprising three subgroups of physical symptoms. Internal consistency was high for the psychological subscale (Cronbach alpha coefficients of 0.85 for the cancer group and 0.77 for the noncancer group) and for the physical subscale groupings, with coefficients ranging between 0.78 to 0.87. The symptom scores were significantly correlated with perceptions of fatigue. These findings show that both seriously ill cancer and noncancer patients experience symptom distress, and that the MSAS seems to be a reliable measure of symptom distress in noncancer patients, as well as with cancer patients. PMID- 12727040 TI - Multicenter study of pain and its management in patients with advanced cancer in Korea. AB - The aim of this study was to evaluate the prevalence, severity, and management of pain in Korean patients with advanced cancer, and to identify the predictors of inadequate management of cancer pain in Korea. From 8 university hospitals, 655 patients with advanced cancer were surveyed. Information concerning analgesics prescribed was acquired from the medical records by the investigator. Physicians, nurses and caregivers were asked to estimate patients' pain. The Korean Brief Pain Inventory and the Barrier Questionnaire were completed by the patients. The Pain Management Index was estimated. Among all patients, 70.8% (464 of 655) reported pain. Among those who had pain, 63.6% (295 of 464) reported pain rated 5 or higher on a 0-10 scale. Thirty-nine percent of the patients had not received any analgesics and 53.2% were not receiving optimal pain management. Although there was a correlation between patients' pain ratings and those of doctors, nurses, and caregivers, there was no significant correlation between patients' ratings and health care providers' ratings at pain levels above moderate intensity. Cancer pain was more poorly managed in advanced cancer than terminal cancer patients (OR:3.20, 95%C.I, 1.83-5.60), in patients with better performance(OR:3.17, 95%C.I, 1.64-6.11), and in those patients whose pain was underestimated by the doctor (OR:2.58, 95%C.I. 1.42-4.69). Despite the high prevalence and severity of pain in cancer patients, the assessment and management of cancer pain were found to be inadequate in Korea. PMID- 12727042 TI - Variations in patients' self-report of pain by treatment setting. AB - Patient reluctance to report pain has been shown to be a primary reason for inadequate pain control among cancer patients. Very little is known about whether patients' self-reports of pain vary by treatment settings. We reviewed 63 medical records of female breast cancer patients who visited two treatment settings on the same day in a tertiary cancer center in the United States. Patients' rating of pain (on a 0 [no pain]-10 scale) were abstracted. Results showed discrepancies regarding patients' self-reports of pain intensity in the two treatment settings. Fifty-one percent of patients' self-report of pain differed between the two treatment settings, with 38% reporting a pain score > or =4 in the outpatient breast clinic and =0 in the outpatient chemotherapy clinic. Although it is expected that pain may vary on a sporadic or activity-related basis, these results may also indicate the need to review clinic staffs' methods of assessing patients about their pain and a review of documentation practices of pain assessment. PMID- 12727041 TI - Antimicrobial use in patients with advanced cancer receiving hospice care. AB - Patients with advanced cancer receiving hospice and palliative care are highly susceptible to infections. The decision whether to treat an active or suspected infection in end-of-life care may be difficult. In order to develop guidelines for the use of antimicrobials (antibiotics and antifungals) in palliative care, we discussed antimicrobial options with 255 patients with advanced cancer at the time they entered a community-based hospice and palliative care program. We subsequently documented the use and effectiveness of the antimicrobials employed during the palliative care period. Most patients (79.2%) chose either no antimicrobials or symptomatic use only. Choices were influenced by age, the desire for symptom control, life-prolongation issues, and the condition of the patient. After admission, 117 patients had a total of 129 infections, with the most common sites being urinary tract, respiratory tract, mouth/pharynx, and skin/subcutaneous tissues. The use of antimicrobials controlled symptoms in the majority of the urinary tract infections, but were less effective in controlling symptoms in the other sites of infection. Survival was not affected by the patients' choice of whether to use antimicrobials, the prevalence of infections, or the actual use of antimicrobials. Symptom control may be the major indication for antimicrobial use for patients receiving hospice and palliative care. PMID- 12727043 TI - Validation of the German version of the brief fatigue inventory. AB - Sedation and tiredness are among the most frequent symptoms among cancer patients. A detailed assessment of these symptoms is necessary to evaluate therapeutic effects, such as the use of methylphenidate or comparison of different opioids. The Brief Fatigue Inventory (BFI) has been validated as a short and comprehensive instrument to assess severity of fatigue and fatigue related impairment in cancer patients. We validated the German version of the BFI in patients with chronic cancer-related and noncancer-related pain treated in a tertiary pain center. Patients treated in the Pain Clinic of the Department of Anesthesiology completed the BFI, the minimal documentation system (MIDOS) and the short form SF-36 quality-of-life questionnaire (SF-36). Test-retest reliability was assessed with a second BFI immediately after the consultation and in a subgroup of patients after 3 to 7 days. Nineteen percent of the 117 patients were treated for cancer-related pain (C); the other patients suffered from chronic severe pain of nonmalignant origin (NC). Patients reported mean values for average fatigue of 3.9 (C) and 4.9 (NC), and for worst fatigue of 5.5 (C) and 6.2 (NC). The mean score of the 6 impairment items was 4.3 in both groups. Factor analysis led to a solution with one common factor for all nine items. Fatigue on the BFI correlated highly with 'feeling tired' in the SF-36 and with 'sedation' in MIDOS, and less with 'being worn out' in SF-36 and 'weakness' in MIDOS. Internal consistency was high, as was test-retest reliability, with a correlation of the intensity, mean scores of 0.93 and the impairment mean scores of 0.87. In conclusion, we found the German version of the BFI to be reliable and valid for cancer and noncancer patients. Minor differences were seen in the validation compared to the original version. PMID- 12727044 TI - Respiratory function after unilateral percutaneous cervical cordotomy. AB - Percutaneous cervical cordotomy (PCC) is a recognized procedure for control of pain due to thoracic malignancies. Caution with PCC in those with precarious lung function has previously been advised. Thirty-five patients were studied in a prospective study of respiratory function before and after PCC for control of pain from pleural mesothelioma or other thoracic malignancy using standard, easily applied tests. Mean duration of survival was 83 days (range 3-360 days). FEV1.0 and FVC did not alter significantly after the procedure. There was no relationship between any of the respiratory function variables measured and survival. Transient nocturnal hypoxemia occurred during the night immediately following PCC in 6 patients. Unilateral PCC does not worsen respiratory function in patients with pleural mesothelioma or other thoracic malignancies. Poor respiratory function before PCC does not predict survival or complications. It should not be a barrier to use of PCC. PMID- 12727045 TI - Stability and compatibility of morphine-clonidine admixtures in an implantable infusion system. AB - Nonopioid analgesics are often coadministered with intrathecal morphine to increase efficacy. The purpose of this study was to evaluate stability and compatibility of morphine-clonidine admixtures with an implantable infusion system that is commonly used to treat pain patients. Infusion systems were filled with admixture and maintained at 37 degrees C for 90 days. Samples were collected monthly. Drug concentrations were determined using stability-indicating, high performance liquid chromatography. For compatibility testing, individual materials comprising the fluid pathway of the device were immersed in clonidine solution and stored at 37 degrees C for various periods through 64 weeks and mechanical performance evaluated. After 3 months of containment in the infusion system, morphine and clonidine concentrations remained at > or = 94% of the theoretical starting concentrations. All device materials retained acceptable mechanical performance following clonidine exposure. These results demonstrate that morphine and clonidine are stable when combined in aqueous solution maintained at body temperature in an implantable infusion system for at least 3 months. PMID- 12727046 TI - Sex differences in pain-related symptoms and self-initiated school nurse visits among pre-adolescents. AB - We examined sex differences in self-initiated school nurse visits and pain associated symptoms in 57 pre-adolescents (36 girls), aged 9-11, with a brief, pain-focused version of the Children's Somatization Inventory (CSI). Using sequential multiple regression, pain symptom scores, sex, and their interaction were analyzed in relation to nurse visits prospectively collected for 1 year. Neither sex, nor pain scores alone, predicted total nurse visits. For girls only, higher pain symptom scores predicted increased total visits and increased visits for complaints with documented physical findings. For both sexes, higher pain symptom scores predicted more visits for complaints without documented physical findings. A complex relationship between sex, pain-related symptoms, and self initiated nurse visits was found in this pre-adolescent sample. Our findings suggest pain-focused symptom measures may be useful in studies of early predictors of adult pain-related healthcare utilization. PMID- 12727047 TI - Rapid progression of advanced "hormone-resistant" prostate cancer during palliative treatment with progestins for cancer cachexia. AB - We report three patients with advanced "hormone-resistant" prostate cancer, each of whom had rapid progression of the disease during treatment with megestrol acetate for cancer cachexia. All patients had been previously treated with total androgenic deprivation. With progression of the disease, megestrol acetate was given to palliate the cancer-related wasting syndrome. No other antineoplastic drugs were contemporaneously given, and no concomitant condition that could favor the progression of the disease was present. The worsening observed while receiving megestrol acetate, and the atypical withdrawal syndrome occurring after the treatment was stopped, seem to suggest a promoting role of megestrol acetate in advanced "hormone-resistant" prostate cancer. The risk of rapid disease progression overwhelming the anti-cachectic palliative effect should be kept in mind when progestins are administered as a palliative treatment of cancer cachexia in patients with advanced "hormone-resistant" prostate cancer. PMID- 12727048 TI - A retrospective chart review of the use of olanzapine for the prevention of delayed emesis in cancer patients. AB - Chemotherapy-induced delayed emesis (DE) affects approximately 50-70% of patients receiving moderately and highly emetogenic chemotherapy. DE most commonly occurs within the first 24-48 hours of chemotherapy administration and can persist for 2 5 days. Olanzapine, which has been used anecdotally for chronic nausea in advanced cancer patients, might be a useful treatment for the prevention of delayed emesis in chemotherapy patients. We conducted a chart review to explore this hypothesis and to plan potential studies. Using pharmacy records or an electronic medical record, we identified all patients who had received olanzapine in the oncology clinic (n = 98). We reviewed these records and selected all patients (n = 28) who had received olanzapine for the prevention of delayed emesis for structured review. There were 17 women (60.7%) and 11 men (39.3%). Eleven patients (39.3%) had at least one instance of nausea recorded while undergoing olanzapine treatment and seven (25%) had an episode of vomiting recorded. During 95 total cycles of chemotherapy with olanzapine (mean = 3.4 cycles per patient), there were 21 incidents of nausea (22.1%) and 10 instances of vomiting (10.5%). Side effects were rarely noted. These data suggest that olanzapine was well tolerated and may reduce the incidence of delayed emesis in patients receiving moderate to highly emetogenic chemotherapy. A series of prospective trials are underway. PMID- 12727049 TI - Rapid proton fat-water spectroscopy for the characterization of non-CNS lesions in vivo. AB - The presence or absence of fat in lesions can have important diagnostic implications. Current MR techniques for the evaluation of fat within lesions in the body rely on indirect imaging methods. The goal of this study was to develop a rapid clinically practical proton spectroscopy procedure for the direct observation of a localized fat-water signal within the body. The technique developed reliably determined fat-water ratios in phantoms and from lesions in vivo in 6 s with single voxel sizes as small as 0.125 cc. PMID- 12727050 TI - Fatty tissue breast lesions. AB - Hamartoma, lipoma and fat necrosis are benign fatty tissue lesions that may present as breast lumps. The mammographic appearance may be distinctive allowing imaging diagnosis without biopsy. The characteristic feature of hamartoma is that of a compressible mass containing radiolucent fat interchanged with dense fibrous connective tissue surrounded by a thin radioopaque pseudocapsule. Lipomas are radiolucent with well-defined thin smooth capsule. Twenty-seven percent of fat necrosis appears as discrete round or oval radiolucent oil cyst with thin capsule and egg-shell calcification. PMID- 12727051 TI - The challenge of the solitary pulmonary nodule: diagnostic assessment with multislice spiral CT. AB - The advent of fast multiscale computed tomography (MSCT) technology has sparked new interest in the noninvasive assessment of the solitary pulmonary nodule (SPN). Fast scanning within a single breath-hold period, simultaneous acquisition of multiple thin slices with subsequent morphologic characterization of the nodule, determination of perfusion patterns as well as growth rates has led to unprecedented improvements in this emerging field. This article reviews the capabilities of MSCT in the diagnostic assessment of the SPN. PMID- 12727052 TI - A case of pyothorax-associated lymphoma simulating empyema necessitatis. AB - We describe a case of a diabetic man with a 40-year history of chronic tuberculous empyema presented with fever, chest pain and bulging soft tissue of the right chest wall. CT scan revealed a huge chest wall mass showing extensive necrosis with air-bubbles and destruction of the ribs. Decortication and extirpation of the chest wall mass were performed, and histopathologic examination confirmed diffused large cell type non-Hodgkin's lymphoma. PMID- 12727053 TI - Lymphangioleiomyomatosis initially presenting with abdominal pain: a case report. AB - Lymphangioleiomyomatosis (LAM) is a rare idiopathic disease characterized by hamartomatous smooth muscle proliferation of the lymph node, lymphatics, blood vessels as well as airways within the lungs, mediastinum and abdomen. It exclusively affects women during the reproductive period. Though extrapulmonary manifestations have been reported, the initial presentation of LAM as abdominal pain is extremely rare. To our knowledge, there are only three cases with LAM presenting first with abdominal symptoms so far [Chest 106 (1994) 267; Eur J Radiol 14 (1992) 192; Eur J Surg 157 (1991) 36]. We describe a case of LAM suffering from abdominal pain followed by pulmonary symptoms and the diagnosis was not made until pathohistological examination. PMID- 12727054 TI - Colour duplex ultrasound-guided sclerotherapy: a new approach to the management of patients with peripheral vascular malformations. AB - INTRODUCTION: Colour duplex ultrasound (CDU)-guided sclerotherapy is a valuable modality for treating peripheral vascular malformations (PVMs). PATIENTS AND METHODS: Between April 1996 and October 1998, 75 patients (age 5-65 years) with PVMs were subjected to CDU. Sclerotherapy was subsequently performed on 40 patients, without sedation, using 3% sodium tetradecyl sulfate, with mean follow up of 4 years. RESULTS: CDU of the 33 high-flow lesions (HFLs) revealed direct arteriovenous (AV) communicating channels with very high forward diastolic flow in seven lesions (arteriovenous fistulas, AVFs), but not in the other 25 lesions (non-AVF). One was a mixed lesion picked up by CDU. Sixteen HFLs were subjected to sclerotherapy; 13 (81.25%) regressed. CDU of the 42 low-flow lesions (LFLs) helped categorize them into Type 1, where no supplying arteries could be seen (12 lesions), and Type 2, where supplying arteries were seen (30 lesions). Type 2 lesions could be further subcategorized based on the spectral trace of their supplying arteries: Type 2a, high-resistance flow (25 lesions); and Type 2b, low resistance flow with a small forward diastolic flow (5 lesions). Twenty-four LFLs were subjected to sclerotherapy; 20 (83.3%) regressed. CONCLUSION: CDU findings correlated well with the clinical appearances of PVMs, and helped to further subcategorize these lesions based on flow. Significant differences in the Doppler flowmetry parameters of the supplying arteries seen in the HFLs and LFLs have enabled us to suggest values for differentiating between them. CDU was also found to be valuable in the follow-up of these lesions. PMID- 12727055 TI - Sonography and venography of the lower extremities for diagnosing deep vein thrombosis in symptomatic patients. AB - The purpose of this study was to investigate the efficacy of sonography and the frequency of indeterminate sonographic examinations in the evaluation of patients with suspected lower extremity deep vein thrombosis (DVT). We prospectively evaluated 136 symptomatic patients (157 extremities) with suspected DVT using sonography and contrast-enhanced venography (n=106 patients, 115 extremities). Using venography as the reference standard for diagnosing DVT, the sensitivity and specificity of sonography was 92.8% and 98%, respectively, yielding an accuracy of 96.8%. The frequency of indeterminate examinations for calf DVT was 32.4%. One (0.7%) fatal pulmonary embolus occurred in our patients. The pulmonary embolism (PE) rate was 1.6% after lower extremity sonography with negative results. Sonography is highly accurate in detecting lower extremity DVT in symptomatic patients. Because of the high frequency of indeterminate studies in the calf and the associated possible risk of pulmonary emboli, we urge radiologists exercise additional caution when evaluating symptomatic patients with clinically suspected lower extremity DVT. PMID- 12727056 TI - Angiographic findings in tumoral calcinosis. AB - Tumor calcinosis is uncommon, typically manifesting as paraarticular, extracapsular soft tissue deposits containing amorphous calcium phosphate and calcium carbonate, with associated hydroxyapatite crystal. CT and MRI are the primary diagnostic radiological tools evaluating these lesions. Primary treatment is early surgical excision with wide margins, as there is a high recurrence rate. We describe the angiographic findings in tumoral calcinosis, demonstrating hypervascularity beyond the calcified mass periphery. Exact margin definition with angiography may influence management and surgical approach. PMID- 12727057 TI - Extraskeletal mesenchymal chondrosarcoma: case report. AB - This is a case report of an extraskeletal mesenchymal chondrosarcoma (ESMC) that originally occurred in the retroperitoneum of a 24-year-old female and later metastasized to the left proximal humerus. Mesenchymal chondrosarcomas are very rare in comparison to conventional chondrosarcomas and even more so when arising from an extraskeletal location. In this report, we discuss the major characteristics of ESMC and offer a review of the current knowledge regarding this rare disease entity. PMID- 12727058 TI - Sciatic schwannoma of the thigh detected on bone scan: a case report. AB - Sciatic nerve schwannomas are rare and may present diagnostic difficulties. This case was incidentally detected on a three-phase bone scan to rule out reflex sympathetic dystrophy (RSD) of the right foot. PMID- 12727059 TI - MR arthrography in chondromalacia patellae diagnosis on a low-field open magnet system. AB - OBJECTIVE: The purpose of this study was to compare the diagnostic efficacy conventional MRI and MR arthrography (MRA) in the diagnosis of chondromalacia patella (CP) on a low-field open magnet system (LFOMS), correlated with arthroscopy. SUBJECTS AND METHODS: Forty-two patients (50 knees) with pain in the anterior part of the knee were prospectively examined with LFOMS, including T1 weighted, proton density-weighted and T2-weighted sequences. All were also examined T1-weighted MRI after intraarticular injection of dilue gadopentetate dimeglumine. Two observers, who reached a consensus interpretation, evaluated each imaging technique independently. Thirty-six of the 50 facets examined had chondromalacia shown by arthroscopy, which was used as the standard of reference. The sensitivity, specificity and accuracy of each imaging technique in the diagnosis of each stage of CP were determined and compared by using the McNemar two-tailed analysis. RESULTS: Arthroscopy showed that 16 facets were normal. Four (30%) of 13 grade 1 lesions were detected with T1. Four lesions (30%) with T2 and three lesions (23%) with proton-weighted images were detected. Seven (53%) of 13 grade 1 lesions were detected with MRA. Grade 2 abnormalities were diagnosed in two (33%) of six facets with proton density-weighted pulse sequences, two (33%) of six facets with T1-weighted pulse sequences, in three (50%) of six facets with T2-weighted pulse sequences, in five (83%) of six facets with MRA sequences. Grade 3 abnormalities were diagnosed in three (71%) of seven facets with proton density- and T1-weighted images, five (71%) of seven facets with T2-weighted pulse sequences, six (85%) of seven facets with MRA sequences. Grade 4 CP was detected with equal sensitivity with T1-, proton density- and T2-weighted pulse sequences, all showing seven (87%) of the eight lesions. MRA again showed these findings in all eight patients. All imaging techniques were insensitive to grade 1 lesions and highly sensitive to grade 4 lesion, so that no significant difference among the techniques could be shown. CONCLUSION: All imaging technique studied had high specificity and accuracy in the detection and grading of CP; however, MRA was more sensitive than T1-weighted and proton density-weighted MR imaging on a LFOMS. Although the arthrographic techniques were not significantly better than T2-weighted imaging, the number of false-positive diagnosis was greatest with T2-weighted MRI. PMID- 12727060 TI - Desmoplastic fibroma of bone: case report. AB - Desmoplastic fibroma of bone is an exceedingly rare tumor that was first described by Jaffe in 1958 [1]. It has a high incidence of local recurrence after surgical resection. It is usually seen in young patients and involves mandible and long bones. It has histological resemblance to the desmoid tumor of the abdominal wall. We report a surgically proven case of desmoplastic fibroma of bone with local aggressiveness and recurrence during follow-up. The radiograph, CT imaging features, radiological and pathological differential diagnosis of the case are described, and literature is briefly reviewed. PMID- 12727061 TI - Mixed sclerosing bone dysplasia--a case report with literature review. AB - We report a case of mixed sclerosing bone dysplasia in a 26-year-old man. This is a very rare disorder characterized by a variable combination of melorheostosis, osteopoikilosis and osteopathia striata. The disease may be generalized or may show unilateral involvement. PMID- 12727062 TI - Primary leiomyosarcoma of the pulmonary artery: a diagnostic dilemma. AB - Primary leiomyosarcoma of the pulmonary artery is a rare malignancy arising from the multipotential mesenchymal cell of the intima of the pulmonary artery. Due to its rarity and nonspecific clinical symptoms, the correct diagnosis and proper management are often delayed. Furthermore, it is frequently misdiagnosed as pulmonary embolism, mediastinal mass, pulmonary stenosis and lung cancer. Therefore, it is important to consider primary leiomyosarcoma of the pulmonary artery a possibility when a persistent filling defect is present in the pulmonary artery and there is no response to optimal anticoagulation treatment. Radiologic findings such as a unilateral mass continuously filling the pulmonary artery, inhomogenous enhancement, vascular distension, extravascular invasion into adjacent structure or uptake in the area of tumor on the FDG-PET can be helpful when differentiating pulmonary artery sarcoma (PAS) from chronic thromboembolism. PMID- 12727070 TI - Use of imipenem as empirical treatment of febrile neutropenia. AB - Based on past information from the literature and our own review, it appears that imipenem can be used effectively as an initial empirical therapy of febrile neutropenia, as a monotherapy, even in patients with haematological malignancies. The response rate is outstanding in microbiologically documented infections, namely bacteraemias, although more information is needed about its optimal use when pneumonia is present. There is no logical or medicine-based evidence suggesting that imipenem should be reserved for second line therapy in patients with febrile neutropenia not responding to empirical treatments that did not contain imipenem. A lower dose of imipenem (500 mg every 6 h) is probably as effective and definitely better tolerated, than higher doses, especially as far as nausea and vomiting are concerned. PMID- 12727071 TI - Molecular analysis of rifampicin-resistant Mycobacterium tuberculosis isolated in Spain (1996-2001). Description of new mutations in the rpoB gene and review of the literature. AB - Mutations of the 81-bp-core region of rpoB gene associated with rifampicin resistance were studied in 169 clinical strains of Mycobacterium tuberculosis isolated from Spain between 1996 and 2001. The analysis identified 16 single base changes, one deletion, two insertions and five multiple mutations (with two or three codons implicated). Eight strains had no mutations, although they were resistant by the proportion method. The five single nucleotide changes with two or three codons implicated and one insertion have not been previously reported. The analysis of the RFLP-IS6110 pattern showed a great heterogeneity (159 individual patterns and four patterns with two or three strains) that suggests no geographical or temporal relations between the studied strains. The concordance rate of InnoLipa Rif-Tb probe with sequencing and phenotypic methods was 94.6%. The results indicate that the InnoLipa probe assay may be useful for rapid screening of the most frequent mutations found in rifampicin-resistant M. tuberculosis strains. PMID- 12727072 TI - Mutations in GyrA, ParC, MexR and NfxB in clinical isolates of Pseudomonas aeruginosa. AB - The target enzymes GyrA and ParC and two efflux pump regulatory genes mexR and nfxB were analysed to determine changes associated with fluoroquinolone resistance in Pseudomonas aeruginosa. Both low- and high-level ciprofloxacin resistance was associated with a Thr-83Ile substitution in GyrA. A ParC Ser-80Leu substitution was found in highly resistant isolates in tandem with the Thr-83Ile substitution in GyrA. Mutations in the efflux regulatory genes were associated with resistance only when in tandem with a mutation in GyrA or ParC. These data show that the main mechanism of fluoroquinolone resistance in P. aeruginosa is mediated primarily through mutations in GyrA, and that mutations in ParC and the efflux regulatory genes are secondary. PMID- 12727073 TI - Mutations causing in vitro resistance to azithromycin in Neisseria gonorrhoeae. AB - In 1999, a cluster of gonococcal isolates exhibiting high Minimal Inhibitory Concentrations (MICs), to azithromycin (2.0-4.0 mg/l) were identified in Kansas City, MO. Isolates were characterized by auxotype/serovar class, lipoprotein (Lip) subtyping and sequencing of the mtrR gene, which has been implicated in decreased azithromycin susceptibility in the gonococcus. Isolates were Pro/IB-3 and contained the 17c Lip subtype. Molecular characterization of the mtrR gene revealed a 153 base pair insertion sequence located between the mtrR/mtrC promoter and the mtrC gene. Some isolates also contained a frame shift within the mtrR gene. Transformation of these mutations into an azithromycin-sensitive recipient strain resulted in transformants with MICs as high as 2.0 mg/l and inactivation of the mtrD gene reduced azithromycin MICs 270-fold. These results demonstrated that the mtr mutations were responsible for the increased MICs in these isolates. PMID- 12727074 TI - Emergence of phenotypic resistance to ciprofloxacin and levofloxacin in methicillin-resistant and methicillin-sensitive Staphylococcus aureus strains. AB - The emergence of phenotypic resistance to ciprofloxacin and levofloxacin in methicillin-sensitive and methicillin-resistant Staphylococcus aureus (MRSA) strains was studied. Twenty MRSA and 77 methicillin-sensitive S.aureus (MSSA) strains susceptible to both quinolones were investigated for resistance after single step or serial passages. No growth of 20 MRSA strains was observed at 4xMIC of levofloxacin after 48 h incubation, but 4 of 77 (5%) MSSA strains grew at the same concentration. At 4xMIC concentration of ciprofloxacin, 10 MSSA (13%) and five MRSA (25%) strains were grown. In the serial passages of MRSA strains, resistance to ciprofloxacin was 75 and 5% for levofloxacin by the third passage. In the seventh passage this resistance was 100 and 15%, respectively. In MSSA strains, resistance to ciprofloxacin was 75 and 19% to levofloxacin at the third passage and at the seventh passage, 100 and 61%, respectively. Emergence of ciprofloxacin resistance was more common and developed more rapidly than resistance to levofloxacin in both MRSA and MSSA strains. PMID- 12727075 TI - Comparative in vitro potency of amoxycillin-clavulanic acid and four oral agents against recent North American clinical isolates from a global surveillance study. AB - The in vitro activity of amoxycillin-clavulanic acid was compared with four comparator oral antimicrobial agents; ampicillin, azithromycin, cefuroxime and trimethoprim-sulphamethoxazole against 4536 recent clinical isolates covering 29 species isolated in the US and Canada between 1997 and 1999. Based upon Minimum inhibitory concentrations (MICs), amoxycillin-clavulanic acid was the most active agent against many Gram-positive species and phenotypes including methicillin susceptible Staphylococcus aureus (MSSA) Staphylococcus epidermidis, Enterococcus faecalis, Streptococcus pyogenes, Streptococcus pneumoniae including penicillin intermediate and macrolide resistant strains and was as active as ampicillin against Streptococcus agalactiae, penicillin resistant S. pneumoniae and viridans streptococci. Against Enterobacteriaceae amoxycillin-clavulanic acid in general, displayed weak activity with only Proteus mirabilis and Proteus vulgaris displaying levels of susceptibility above the 90th percentile. Amoxycillin clavulanic acid had significant activity against many species of Gram-negative non-Enterobacteriaceae including Haemophilus influenzae, Haemophilus parainfluenzae and Moraxella catarrhalis but negligible activity against Burkholderia cepacia, Pseudomonas aeruginosa and Stenotrophomonas maltophilia. Amoxycillin-clavulanic acid continues to retain excellent activity against the majority of targeted pathogens despite 20 years of clinical use. PMID- 12727076 TI - Susceptibilities of different Actinobacillus actinomycetemcomitans strains to lactoperoxidase-iodide-hydrogen peroxide combination and different antibiotics. AB - Actinobacillus actinomycetemcomitans has an important aetiological role in localized juvenile periodontitis and in progressive periodontitis in adults. A. actinomycetemcomitans is found mainly in periodontal pockets but also in whole saliva, a potential transmission medium. It is sensitive to peroxidase-halide systems, but the differences between periodontitis associated clinical isolates and type strains are unclear. The sensitivities of these 2 strain groups to lactoperoxidase (LP)-iodide (I(-))-hydrogen peroxide (H(2)O(2)) combinations were investigated, and the sensitivities were compared with the susceptibilities to four antibiotics. There was great variation between the sensitivities of different strains, but the 2 strain groups responded similarly. The LP (75 microg)-I(-) (100 nmol)-H(2)O(2) (1000 nmol) combination produced a similar degree of inhibition as 2 microg ampicillin. The LP-I(-) system might be a potential antimicrobial agent against A. actinomycetemcomitans transmission via saliva. PMID- 12727077 TI - Telithromycin (HMR 3647) achieves high and sustained concentrations in tonsils of patients undergoing tonsillectomy. AB - Telithromycin, the first ketolide antimicrobial to be developed for clinical use, has potent activity against group A beta-haemolytic streptococci (GABHS), including macrolide-resistant strains. The penetration of telithromycin into tonsils was assessed in 22 adults undergoing tonsillectomy at 3, 12 or 24 h after the fourth dose of oral telithromycin 800 mg once daily. Telithromycin rapidly penetrated tonsillar tissues, achieving a mean concentration of 3.95 mg/kg at 3 h post dose, 3.4 times greater than the corresponding plasma concentration (1.22 mg/l. The mean tonsil:plasma concentration ratio increased to 13.1 at 24 h post dose, indicating slower elimination from tonsils than plasma. Tonsillar and plasma concentrations exceeded the MIC(50) for GABHS throughout the 24-h dosing period. These findings suggest that telithromycin may be an effective new alternative treatment for GABHS tonsillopharyngitis. PMID- 12727078 TI - In vitro pharmacodynamic properties of a fluoroquinolone pharmaceutical derivative: hydrochloride of ciprofloxacin-aluminium complex. AB - Some in vitro pharmacodynamic properties of a new aqueous soluble ciprofloxacin (CIPX) derivative, the hydrochloride of its aluminum complex: (HCl.CIPX)(3)Al, are reported. Although (HCl.CIPX)(3)Al had the same MIC as CIPX, the minimum bactericidal activity against Escherichia coli was 2-fold higher than that of CIPX and the rate of killing was slightly delayed compared with time-kill curves obtained with CIPX. (HCl.CIPX)(3)Al showed a longer post-antibiotic effect (PAE). As pharmacodynamic properties of CIPX are not drastically affected by being complexed with aluminium, the increased aqueous compatibility of the complex remains as the main formulation factor for liquid dosage forms. PMID- 12727079 TI - Does dexamethasone affect ceftriaxone [corrected] penetration into cerebrospinal fluid in adult bacterial meningitis. AB - Trough cerebrospinal fluid (CSF) ceftriaxone concentrations were measured daily to investigate the effect of dexamethasone on ceftriaxone penetration into CSF in adult patients with acute bacterial meningitis. Patients were divided into two groups in this double blind randomized study. In group 1 (n=6) patients were given ceftriaxone with dexamethasone whereas in group 2 (n=6) patients were only administered ceftriaxone. Plasma and CSF samples were collected at 24, 48, 72, 96 and 264 h following the study treatments. The trough CSF ceftriaxone concentrations were measured using high performance liquid chromatography (HPLC) and microbiological assay. CSF ceftriaxone concentrations were 3.21 mg/l at 24 h in group 1 and 4.85 mg/l at the same time in group 2 by HPLC. Although microbiological assay results were lower than HPLC the trough CSF ceftriaxone concentrations in dexamethasone group were at least 10(3) times higher than the minimum inhibitory concentrations of the susceptible strains. It was concluded that the ceftriaxone concentration in CSF was adequate and ceftriaxone penetration was not significantly affected by concomitant dexamethasone use in adult patients with acute bacterial meningitis. PMID- 12727080 TI - Comparative analysis of azithromycin and ciprofloxacin in the treatment of chronic prostatitis caused by Chlamydia trachomatis. AB - A total of 89 patients, (>18 years), with symptoms of chronic prostatitis and inflammatory findings as well as the presence of Chlamydia trachomatis confirmed by DNA/RNA DIGENE hybridization method and/or by isolation, McCoy culture and Lugol stain in expressed prostatic secretion or in voided bladder urine collected immediately after prostatic massage, were examined. The patients were randomized to receive a total of 4.5 g of azithromycin for 3 weeks, given as a 3-day therapy of 1 x 500 mg weekly or ciprofloxacin 500 mg b.i.d. for 20 days. Patients' sexual partners were treated at the same time. Clinical and bacteriological efficacy were evaluated 4-6 weeks after the end of therapy. Significantly higher eradication (36/45: 17/44; P=0.0002) and a significantly higher clinical cure (31/45: 15/44; P=0.0021) were achieved in the group of patients treated with azithromycin than in the ciprofloxacin group. PMID- 12727081 TI - Relevance of antimicrobial agent-induced endotoxin release from in vitro cultured Escherichia coli and in vivo experimental infection with Gram-negative bacilli. AB - In vitro exposure of Gram-negative bacilli (GNB) to antimicrobial agents may induce endotoxin (ET) release, that may cause various reactions in vivo resulting in endotoxic shock. We used the antimicrobial agents, flomoxef (FMOX) and gentamicin (GM), to investigate the kinetics of ET released from in-vitro cultured Escherichia coli and to examine the ET effect on tumor necrosis factor (TNF) production by macrophages. In a rabbit model of E. coli peritonitis, we measured plasma ET, TNF and blood bacterial counts under the administration of FMOX or GM. In our in vitro experiment, ET levels under FMOX were significantly higher than those under GM, and ET induced TNF production in a dose-dependent manner. However, in vivo, plasma ET, TNF, and blood bacterial counts under antimicrobial agents were significantly lower than those of the controls, and those under FMOX treatment did not differ from those under GM treatment. Thus, ET release may not be a critical problem in GNB infections if appropriate antimicrobial agents are administered. PMID- 12727082 TI - Treatment with rokitamycin suppresses the lethality in a murine model of Escherichia coli O157:H7 infection. AB - The effects of rokitamycin (ROK) and levofloxacin (LEVX) were investigated in a murine model of enterohaemorrhagic Escherichia coli (EHEC) infection. After C3H/HeN mice were inoculated intragastrically with E. coli O157:H7, ROK (20mg/kg) or LEVX (1.2 mg/kg) was administered intragastrically. The death rate of the mice was noted and the faeces were collected to determine viable cell counts of EHEC and for Shiga-like toxins (SLTs) assays. After the mice were sacrificed, the kidneys and colons of some of the mice were removed for histopathological examination. The death rate of mice administered ROK (19%) was significantly lower than that of the control and LEVX-treated groups (80, 93%, respectively). Viable cell counts of EHEC in the faeces of the control and ROK-treated groups were 10(7) and 10(6) CFU/g at day 5 after the infection, respectively. LEVX reduced the bacterial count by less than 100 CFU/g at day 5. The level of SLTs in the faeces from the ROK group were lower than the LEVX-treated and control groups at day 5. The histopathological findings in the kidneys treated with LEVX showed necrotic tubular epithelial cells and those in the colon, inflammatory infiltrates. These were not seen in the ROK-treated group. These results suggested that ROK suppressed release of SLTs from the EHEC and could be useful in the treatment of EHEC infection. PMID- 12727083 TI - Factors associated with the evolution of the viral load in individuals with HIV infection. AB - The temporal evolution of the viral load (VL) in a group of patients under clinical care and the identification of individual variables that determined this evolution was studied using a retrospective descriptive study of the VL determinations on 1336 patients between September 1996 and June 2000. During follow-up, patients' VLs increasingly fell below the detection threshold. Those patients who had received antiretroviral treatment prior to the monitoring of the viraemia presented lower VLs at the start of their follow-up. Those patients who had been cared for on the same unit throughout their follow-up maintained lower VLs. Female patients uniformly presented lower VL results than the males. The immunological status at the beginning of follow-up and the basal VL determined the initial evolution of the patients' VL, although it did not do so in the final stages of follow-up. The immunological and virological state at the onset of the follow-up determines the evolution of the patients' VL. PMID- 12727084 TI - Shortage of penicillin G: impact on antibiotic prescribing at a US tertiary care centre. AB - A six-year retrospective study was undertaken to examine the effect of the recent penicillin G (penG) shortage on antibiotic prescribing practices at a tertiary care centre in the US. Prior to the shortage, penG was predominantly used for intrapartum prophylaxis of group B streptococcal disease. As the supply of penG ended temporarily in October 1999, penG use declined dramatically, ampicillin replaced penG in obstetrics and a shift to broad-spectrum agents was noted in non obstetric patients potentially eligible for penG treatment. Shortage-triggered treatment changes had a negative effect on prescribing patterns, which may contribute to antibiotic resistance. PMID- 12727085 TI - In vitro susceptibility of Arcobacter butzleri and Arcobacter cryaerophilus to different antimicrobial agents. AB - Seventeen strains of Arcobacter butzleri and thirteen of Arcobacter cryaerophilus, were tested for their antimicrobial susceptibility to 26 antimicrobial agents. Among beta-lactams agents in this study, imipenem was the most active agent against both A. butzleri and A. cryaerophilus isolates with MIC(90) values of 2 and 4 mg/l, respectively. The most active cephalosporin tested was cefepime, although it was more active against A. butzleri (MIC(90) 8 mg/l) than A. cryaerophilus (MIC(90) 64 mg/l). Levofloxacin, marbofloxacin, enrofloxacin and ciprofloxacin were the best-performing fluoroquinolones against these species. Of the aminoglycosides, amikacin was the most active agent against both A. butzleri and A. cryaerophilus strains with MIC(90) values of 64 and 16 mg/l, respectively. All isolates showed high levels of resistance to penicillins, macrolides, chloramphenicol, trimethoprim and vancomycin. PMID- 12727086 TI - Ciprofloxacin and co-trimoxazole resistance and extended spectrum beta-lactamase production in Escherichia coli strains isolated from urinary tract infections. PMID- 12727087 TI - In vitro effectiveness of quinupristin/dalfopristin against Staphylococcus aureus strains. PMID- 12727088 TI - Characterization of Klebsiella pneumoniae clinical strains producing a rare extended-spectrum beta-lactamase (IBC-1). PMID- 12727089 TI - The occurrence of the first two CTX-M-3 and TEM-1 producing isolates of Salmonella enterica serovar Oranienburg in Poland. PMID- 12727090 TI - The baccalaureate degree for entry into practice: it's time for nursing to take a stand. PMID- 12727091 TI - Predictors of academic success in a Bachelor of Nursing course. AB - A longitudinal study of one Bachelor of Nursing course (n=181) was carried out to determine the relationship between academic outcomes and pre-entry qualifications and to discuss the progress and success of students with conventional and non conventional qualifications. A multi-linear regression analysis was used to determine which pre-entry qualification best explained the variation in the course marks. Results identified that the number of GCSE A grades obtained significantly predicted success on the BN course. On the basis of this finding it has been suggested that the new AS levels, introduced as part of the reformed 16 19 curriculum, may have greater predictive value than current A levels since they allow a broader range of study. Students entering the course with non conventional qualifications were found to achieve slightly lower marks throughout the course and had a high rate of attrition. Further research is needed to replicate these findings and to determine whether high grades in any specific subject, for example Biology, improve the predictive value of pre-entry qualifications. PMID- 12727092 TI - Evaluating the use of streaming video to support student learning in a first-year life sciences course for student nurses. AB - Streaming video was used to support the learning of first year student nurses on a Life Sciences module, as one of many innovations designed to increase the range of resources and support available to students. This paper describes the background to this innovation, the procedures adopted and the results of extensive evaluation. The use of streaming video was evaluated in three applications in the module. A total of 656 students used online directed-learning sessions that incorporated streamed video. Just over half of these students actually viewed the video streams. Their feedback showed that 32% found access easy, 59% enjoyed using the resources, and 25% were very confident that they learned from them. Different types of video were used, and embedded in diverse ways, but the results were consistent across the three applications. They suggest that streamed video can contribute to useful resources to support learning by student nurses but, for a variety of reasons, it may not appeal or be adequately accessible to all students at present. PMID- 12727093 TI - Clinical environment as a learning environment: student nurses' perceptions concerning clinical learning experiences. AB - The aim of this study is to describe student nurses' perceptions of clinical learning experiences in the context of the clinical learning environment. We use the phenomenological approach by Colaizzi. The data was collected by unstructured interviews. Altogether 16 student nurses were asked to describe the significance of clinical learning experiences and good and bad learning experiences. Four elements sum up these clinical learning experiences: the appreciation and support the students received, the quality of mentoring and patient care, and students' self-directedness. Student nurses valued clinical practice and the possibilities it offered in the process of growing to become a nurse and a professional. A good clinical learning environment was established through good co-operation between the school and the clinical staff. It was concluded that the school should be able to provide a suitable clinical learning environment at the right time, so that theory and practice would complement each other. The teacher was the expert on nursing education, the aims set for each practice, as well as student nurses and their skills, but then again the nurse mentor knew the ward on which students were practicing. This was why collaboration between nurse mentors and nurse teachers was considered very necessary. PMID- 12727094 TI - From process to outcome: the effect of portfolio assessment on student learning. AB - Literature suggests that assessment may influence student learning in a positive or negative way. Despite the support for the use of portfolio assessment, relatively little is known about its effect on student learning. The purpose of this study was to evaluate the effectiveness of portfolio assessment in enhancing student learning. Students undertaking the Bachelor of Nursing programme in a University in Hong Kong participated in the study. Twelve of the students who underwent portfolio assessment described, in individual semi-structured interviews, their experience and perceptions of this form of assessment. This paper reported on the qualitative findings of the study, which employed both qualitative and quantitative methods. Three themes emerged from the students' accounts: (1) the students favoured the use of portfolio assessment; (2) the process of preparing portfolios yielded positive academic and affective outcomes; and (3) unexpected findings in the form of spontaneous collaborative learning and apparent increased interest in learning during the process of preparing portfolios for those students who lacked motivation. The qualitative findings contribute to a better understanding of the use of portfolio assessment and have implications for teaching. PMID- 12727095 TI - Characteristics [correction of charactersistics] of intercultural mentoring--a mentor perspective. AB - This article reports a study of Finnish preceptors' and British undergraduate nursing students' mentor-student relationship during international placement in Finland from the mentors' perspective. The study aimed to explore the characteristics of intercultural mentorship and the strategies used by the mentors to improve the students' intercultural competence. Altogether 23 mentors and five students participated in this study. The data consisted of mentoring session observations, group interviews and research diary notes. Intercultural mentorship was characterised by concern about the students' adjustment, pervasiveness of the relationship, sense of mutual learning, inadequate school placement co-operation and concern about learning outcomes. The mentors used a variety of strategies to improve the students' intercultural competence. Mentorship was both a rewarding and a frustrating experience. PMID- 12727096 TI - An investigation into the perceptions of primary care practitioners of their education and development needs for communicating with patients who may not be fluent in English. AB - This paper reports on an empirical research project that assessed educational needs of primary care practitioners for communicating with minority ethnic patients who are not fluent in English. The qualitative study was carried out in general practice settings in a northern inner city locality. Discrepancies are highlighted between patient and practitioner understandings, and between educational models and practitioner views. Educational needs are identified to enhance practitioners' specific skills, knowledge of patient perspectives, and self-awareness. Organisational influences on communication are explored. Key arguments are that "transcultural" educational models should be used critically, and education should enable practitioners to integrate their personal coping strategies with structured practice development planning. PMID- 12727097 TI - Devising and establishing the face and content validity of explicit criteria of consultation competence in UK primary care nurses. AB - AIM: The aims of this study were (a) to devise a set of prioritised criteria of consultation competence which primary care nurses need to acquire and (b) to determine the face and content validity of these criteria. METHOD: The criteria of consultation competence as contained in the Leicester Assessment Package (LAP) were modified for use with primary care nurses and sent to a stratified sample of UK primary care nurses (n=1126) to determine their face and content validity. RESULTS: Support for the seven categories of consultation competence varied from 93-98% and for the 39 component competences from 88-98%. There was no consensus for alternative or additional categories or components. CONCLUSIONS; We have established the face and content validity of the derived competences. These are suitable for the teaching and assessment of consultation skills of primary care nurses, at all stages of their professional development, when incorporated within an educational package. PMID- 12727098 TI - Developing postgraduate awards for critical care. AB - This report outlines a qualitative study that explored the academics' perceptions of postgraduate education for critical care. A framework was generated using models of curriculum design. Focus groups and interviews were undertaken to explore four areas within this framework - taught theory/professional practice, advanced practice skills, assessment and delivery. The findings revealed that ethical and legal aspects of role, research, physiology decision-making and practice skills such as defibrillation, first line treatment and patient assessment were considered extremely relevant to such a programme. Practice assessment was considered essential to ensure clinical credibility of the programme and the practitioner. Frameworks were generated to define the nature of advanced critical care practice. As such, work-based learning and interprofessional supervision were seen as key elements of the programme. Finally areas for further research were identified. PMID- 12727103 TI - Targeting AAV vectors. PMID- 12727104 TI - A multitasking oncolytic adenovirus vector. PMID- 12727105 TI - Effects of the negative control region on expression from retroviral LTR. PMID- 12727106 TI - Localized arteriole formation directly adjacent to the site of VEGF-induced angiogenesis in muscle. AB - We have shown previously that implantation of myoblasts constitutively expressing the VEGF-A gene into nonischemic mouse skeletal muscle leads to overgrowth of capillary-like blood vessels and hemangioma formation. These aberrant effects occurred directly at the implantation site. We show here that these regions result from angiogenic capillary growth and involve a change in capillary growth pattern and that smooth muscle-coated vessels similar to arterioles form directly adjacent to the implantation site. Myoblasts genetically engineered to produce VEGF were implanted into mouse leg muscles. Implantation sites were surrounded by a zone of dense capillary-sized vessels, around which was a second zone of muscle containing larger, smooth-muscle-covered vessels but few capillaries, and an outer zone of muscle exhibiting normal capillary density. The lack of capillaries in the middle region suggests that the preexisting capillaries adjacent to the implantation site underwent enlargement and/or fusion and recruited a smooth muscle coat. Capillaries at the implantation site were frequently wrapped around VEGF-producing muscle fibers and were continuous with the circulation and were not observed to include bone-marrow-derived endothelial cells. In contrast with the distant arteriogenesis resulting from VEGF delivery described in previous studies, we report here that highly localized arterioles also form adjacent to the site of delivery. PMID- 12727107 TI - Induction of functional neovascularization by combined VEGF and angiopoietin-1 gene transfer using AAV vectors. AB - Vectors based on the adeno-associated virus (AAV) deliver therapeutic genes to muscle and heart at high efficiency and maintain transgene expression for long periods of time. Here we report about the synergistic effect on blood vessel formation of AAV vectors expressing the 165 aa isoform of vascular endothelial growth factor (VEGF165), a powerful activator of endothelial cells, and of angiopoietin-1 (Ang-1), which is required for vessel maturation. High titer AAV VEGF165 and AAV-Ang-1 vector preparations were injected either alone or in combination in the normoperfused tibialis anterior muscle of rats. Long term expression of VEGF165 determined massive cellular infiltration of the muscle tissues over time, with the formation of a large set of new vessels. Strikingly, some of the cells infiltrating the treated muscles were found positive for markers of activated endothelial precursors (VEGFR-2/KDR and Tie-2) and for c kit, an antigen expressed by pluripotent bone marrow stem cells. Expression of VEGF165 eventually resulted in the formation of structured vessels surrounded by a layer of smooth muscle cells. Presence of these arteriolae correlated with significantly increased blood perfusion in the injected areas. Co-expression of VEGF165 with angiopoietin-1-which did not display angiogenic effect per se remarkably reduced leakage of vessels produced by VEGF165 alone. PMID- 12727109 TI - Packaging of an AAV vector encoding human acid alpha-glucosidase for gene therapy in glycogen storage disease type II with a modified hybrid adenovirus-AAV vector. AB - We have developed an improved method for packaging adeno-associated virus (AAV) vectors with a replication-defective adenovirus-AAV (Ad-AAV) hybrid virus. The AAV vector encoding human acid alpha-glucosidase (hGAA) was cloned into an E1, polymerase/preterminal protein-deleted adenovirus, such that it is packaged as an Ad vector. Importantly, the Ad-AAV hybrid cannot replicate during AAV vector packaging in 293 cells, because of deletion of polymerase/preterminal protein. The residual Ad-AAV in the AAV vector stock was reduced to <1 infectious particle per 10(10) AAV vector particles. These modifications resulted in approximately 30 fold increased packaging of the AAV vector for the hybrid Ad-AAV vector method as compared with standard transfection-only methods. Similarly improved packaging was demonstrated for pseudotyping the AAV vector as AAV6, and for AAV vector packaging with a second Ad-AAV vector encoding canine glucose-6-phosphatase. Liver-targeted delivery of either the Ad-AAV hybrid or AAV vector particles in acid alpha-glucosidase-knockout (GAA-KO) mice revealed secretion of hGAA with the Ad-AAV vector, and sustained secretion of hGAA with an AAV vector in hGAA tolerant GAA-KO mice. Further development of hybrid Ad-AAV vectors could offer distinct advantages for gene therapy in glycogen storage diseases. PMID- 12727108 TI - Lentiviral-mediated gene delivery to synovium: potent intra-articular expression with amplification by inflammation. AB - Clinical translation of gene-based therapies for arthritis could be accelerated by vectors capable of efficient intra-articular gene delivery and long-term transgene expression. Previously, we have shown that lentiviral vectors transduce rat synovium efficiently in vivo. Here, we evaluated the functional capacity of transgene expression provided by lentiviral-mediated gene delivery to the joint. To do this, we measured the ability of a lentiviral vector containing the cDNA for human interleukin-1 receptor antagonist (LV-hIL-1Ra) to suppress intra articular responses to IL-1beta. Groups of rats were injected in one knee with 5 x 10(7) infectious units of LV-IL-1Ra. After 24 h, a range of doses of fibroblasts (3 x 10(3), 10(4), 3 x 10(4), or 10(5) cells) genetically modified to overexpress IL-1beta was injected into both knees. Intra-articular delivery of LV hIL-1Ra strongly prevented swelling in all treated knees, even in those receiving the greatest dose of IL-1beta(+) cells. Cellular infiltration, cartilage erosion, and invasiveness of inflamed synovium were effectively prevented in LV-hIL-1Ra treated knees and were significantly inhibited in contralateral joints. Beneficial effects were also observed systemically in the lentivirus-treated animals. Interestingly, intra-articular expression of the IL-1Ra transgene was found to increase in relation to the number of IL-1beta(+) cells injected. Further experiments using GFP suggest this is due to the proliferation of cells, stably modified by the integrative lentivirus, in response to inflammatory stimulation. PMID- 12727110 TI - GLP-1 gene delivery for the treatment of type 2 diabetes. AB - Glucagon-like peptide-1 (GLP-1) is a potent insulinotrophic hormone, which makes GLP-1 an attractive candidate for the treatment of type 2 diabetes. However, the short plasma half-life of the active forms of GLP-1 poses an obstacle to the sustained delivery of this peptide. In this study, we evaluated the effect of GLP 1 gene delivery both in vitro and in vivo using a new plasmid constructed with a modified GLP-1 (7-37) cDNA. This cDNA contains a furin cleavage site between the start codon and the GLP-1 coding region. The expression of the GLP-1 gene was driven by a chicken beta-actin promoter (pbetaGLP1). The level of the GLP-1 mRNA was evaluated by RT-PCR 24 h after transfection. The in vitro results showed a dose-dependent expression of GLP-1. Coculture assay of the GLP-1 plasmid transfected cells with isolated rat islet cells demonstrated that GLP-1 increased insulin secretion by twofold, compared to controls during a hyperglycemic challenge. A single injection of polyethyleneimine/pbetaGLP1 complex into ZDF rats resulted in increasing insulin secretion and decreasing blood glucose level that was maintained for 2 weeks. This GLP-1 gene delivery system may provide an effective and safe treatment modality for type 2 diabetes. PMID- 12727111 TI - Hearing and hair cells are protected by adenoviral gene therapy with TGF-beta1 and GDNF. AB - Glial cell line-derived neurotrophic factor (GDNF) overexpression in the inner ear can protect hair cells against degeneration induced by aminoglycoside ototoxicity. The protective efficiency of GDNF increases when it is combined with co-factors such as transforming growth factor beta1 (TGF-beta1), a ubiquitous cytokine. The aim of this study was to determine whether TGF-beta1 receptors are expressed in the inner ear and whether a cocktail of GDNF and TGF-beta1 transgenes provides enhanced protection of the inner ear against ototoxic trauma. Using RT-PCR analysis, we determined that both TGF-beta1 receptors, type 1 and 2 are present in rat cochlea. We co-inoculated two adenoviral vectors, one encoding human TGF-beta1 gene (Ad.TGF-beta1) and the other encoding human GDNF gene (Ad.GDNF) into guinea pig cochleae 4 days prior to injecting an ototoxic dose of aminoglycosides. Inoculated ears had better hearing and fewer missing inner hair cells after exposure to the aminoglycoside ototoxicity, as compared with controls and ears treated only with Ad.GDNF. Cochleae with TGF-beta1 overexpression exhibited fibrosis in the scala tympani regardless of the presence of GDNF. Our results suggest that the adenovirus-mediated overexpression of GDNF and TGF-beta1 can be used in combination to protect cochlear hair cells and hearing from ototoxic trauma. PMID- 12727112 TI - Regulated expression of erythropoietin from an AAV vector safely improves the anemia of beta-thalassemia in a mouse model. AB - In vivo gene transfer is being considered in the systemic delivery of therapeutic proteins. This report evaluates the use of AAV vectors administered into muscle to deliver erythropoietin (Epo) for the treatment of anemia in a mouse model of beta-thalassemia. Injection of vector expressing Epo from a constitutive promoter resulted in Epo overproduction and improved erythropoiesis. However, severe and lethal polycythemia developed. In order to titrate the expression of Epo to therapeutic and non-toxic levels, vectors were constructed to allow pharmacologic control of Epo transcription. Specifically, expression of Epo was dependent on the presence of a chimeric transcription factor that is activated by the orally available small molecule drug rapamycin. beta-thalassemic mice injected with vectors containing the regulated Epo gene failed to show any effect until they were administered a regimen of rapamycin, which led to the production of Epo and an increase in hematocrit values. Epo expression and its hematologic consequences were directly dependent on the dose of rapamycin and were completely reversed when rapamycin was withdrawn. The increase in hematocrit was associated with partial improvements in the abnormalities of red blood cell morphology. This study confirms the value of pharmacologic regulation of transgene expression in the development of safe and effective gene therapies in which biologically active secreted proteins are produced. PMID- 12727113 TI - Induction of specific antitumor immunity in the mouse with the electrofusion product of tumor cells and dendritic cells. AB - Dendritic cells (DCs) are potent antigen-presenting cells capable of inducing primary T-cell responses. Several immunotherapy treatment strategies involve manipulation of DCs, both in vivo and ex vivo, to promote the immunogenic presentation of tumor-associated antigens. In this study, an electrofusion protocol was developed to induce fusion between tumor cells and allogeneic bone marrow-derived DCs. Preimmunization with irradiated electrofusion product was found to provide partial to complete protection from tumor challenge in the murine Renca renal cell carcinoma model and the B16 and M3 melanoma models. Vaccinated survivors developed specific immunological memory and were able to reject a subsequent rechallenge with the same tumor cells but not a syngeneic unrelated tumor line. Antitumor protection in the B16 model was accompanied by the development of a polyclonal cytotoxic T-lymphocyte response against defined melanoma-associated antigens. The therapeutic potential of this type of approach was suggested by the ability of a Renca-DC electrofusion product to induce tumor rejection in a substantial percentage of hosts (60%) bearing pre-established tumor cells. These results indicate that treatment with electrofused tumor cells and allogeneic DCs is capable of inducing a potent antitumor response and could conceivably be applied to a wide range of cancer indications for which tumor associated antigens have not been identified. PMID- 12727114 TI - Prevention of the initial host immuno-inflammatory response determines the long term survival of encapsulated myoblasts genetically engineered for erythropoietin delivery. AB - The present study investigates the respective roles of both the host immune response and the metabolic requirements in determining the long-term survival of erythropoietin-secreting myoblasts within encapsulating polymer membranes. Hollow fiber capsules loaded with a high density of erythropoietin-secreting C(2)C(12) myoblasts survived poorly in the subcutaneous tissue of syngeneic mice, inducing variable hematocrit responses. To determine the role and the nature of the host response, recipients were treated with anti-inflammatory (diclofenac) and immunosuppressive (dexamethasone, FK506) agents. Only immunosuppressive drugs led to improved graft survival after 5 weeks of implantation, indicating an immune process as the cause of cell death. Furthermore, transient blocking of this process allowed long-term preservation of the implanted cells (> 100 days). The formation of necrotic cell cores inside densely packed devices elicited a local chronic inflammatory reaction. Hence, implants were designed to limit early cell death by inserting a supporting matrix and decreasing the number of loaded cells. The most efficient erythropoietin delivery was obtained with matrix-containing capsules that had received the lowest myoblast density. These results highlight the critical role of initial engraftment in the long-term acceptance of encapsulated myoblasts and the need to limit early cell death in the device to prevent subsequent host immuno-inflammatory responses. PMID- 12727116 TI - Multigene expression from a replicating adenovirus using native viral promoters. AB - We have developed a novel therapeutic gene delivery system for oncolytic adenoviruses that takes advantage of the endogenous gene expression machinery (promoters, splicing, polyadenylation signals) of the E3 transcription unit for gene delivery. In this work, we use two sites in the E3 region (6.7 K/gp19K and ADP sites) to demonstrate that (1) multiple therapeutic genes (MCP-3, TNFalpha) can be expressed from a single replicating Ad, (2) timing of expression of these therapeutic genes mimics that of the E3 region genes they replaced, (3) expression of the remaining genes in the complex E3 transcription unit is maintained, and (4) the multigene-expressing virus retains replication competence and ability to induce classical adenovirus cytopathic effects that parallel those of the parental adenovirus (ONYX-320). This system conserves the DNA packaging capacity of the size-constrained viral genome for therapeutic genes and can potentially be used to link therapeutic transgene expression to tumor-restricted viral replication. Potential clinical implications are discussed. PMID- 12727115 TI - RGD inclusion in VP3 provides adeno-associated virus type 2 (AAV2)-based vectors with a heparan sulfate-independent cell entry mechanism. AB - Recombinant adeno-associated virus (AAV) has become an attractive vector system for a number of gene therapy paradigms. However, the utility of AAV vectors is often limited by the absence of heparan sulfate proteoglycan (HSPG), the virus's primary attachment receptor, on the desired target cell population. In order to achieve HSPG-independent gene delivery, several groups have shown that the endogenous tropism of AAV can be expand by genetically altering the viral capsid. However, the parameters of this developing technology have yet to be defined and it has not yet been determined if these modified vectors actually infect cells via these engineered interactions. Previously we constructed a series of insertion mutants spanning the AAV capsid protein gene and identified specific sites that can tolerate the insertion of small exogenous peptides. Here we describe a number of sites within the AAV capsid gene that can be used for the insertion of integrin-targeting peptide epitopes. Incorporation of an Arg-Gly-Asp (RGD)-containing peptide at these sites enables AAV to infect integrin-expressing cells independent of HSPG. Mutant AAV vectors displaying these peptide ligands can be produced to wild-type titer and have been shown to specifically interact with the targeted integrin receptors and mediate infection via this interaction. We report significant increases in gene transfer to Raji, K562, and SKOV-3 cell lines that express integrin, but little HSPG, suggesting that rAAV vectors displaying RGD peptides may be of great utility for treatment of neoplasms characterized by the deficiency of HSPG expression. We have also demonstrated that due to their expanded tropism, these novel vectors are capable of efficient transduction of AAV2-resistant tumors in vivo suggesting that they may offer significant therapeutic advantages. PMID- 12727117 TI - Inhibition of tumor growth by DT-A expressed under the control of IGF2 P3 and P4 promoter sequences. AB - The human IGF2 P3 and P4 promoters are highly active in a variety of human cancers. We here present an approach for patient oriented therapy of TCC bladder carcinoma by driving the diphtheria toxin A-chain (DT-A) expression under the control of the IGF2 P3 and P4 promoter regulatory sequences. High levels of IGF2 mRNA expression from P3, P4 or both promoters were detected in 18 TCC samples (n = 29) by ISH or RT-PCR. Normal bladder samples (n = 4) showed no expression from either promoter. The activity and specificity of the IGF2 P3 and P4 regulatory sequences were established in human carcinoma cell lines by means of luciferase reporter gene assay. These sequences were used to design DT-A expressing, therapeutic vectors (P3-DT-A and P4-DT-A). The activity of both was determined in cell lines (in vitro) and the activity of P3-DT-A was determined in a heterotopic animal model (in vivo). The treated cell lines highly responded to the treatment in a dose-response manner, and the growth rate of the developed tumors in vivo was highly inhibited (70%) after intratumoraly injection with P3-DT-A compared to non-treated tumors (P < 0.0002) or tumors treated by luciferase gene expressing LucP3 vector (P < 0.002). PMID- 12727118 TI - The effects of cyclic stretch on gene transfer in alveolar epithelial cells. AB - Cyclic stretch has been shown to alter cell physiology, cytoskeletal structure, signal transduction, and gene expression in a variety of cell types. To determine the effects of stretch on the gene transfer process, we compared the transfection efficiencies of human A549 cells grown either statically or exposed to 10% cyclic stretch (Delta surface area) at 60 cycles/min (1 Hz) for 24 hours prior to, and/or after transfection with pEGFP-N1 and pCMV-lux-DTS using lipoplex or electroporation. Stretching the cells prior to transfection had no effect on gene transfer. By contrast, cyclic, but not continuous, stretch applied immediately after transfection for as little as 30 minutes resulted in a 10-fold increase in gene transfer and expression by either transfection technique. These stretch conditions did not result in rupture of the plasma membrane based on the fact that DNA was unable to enter stretched cells unless either an electric field was applied or the DNA was complexed with liposomes. Taken together with the timing of the stretch response and the known effects of stretch on transcription, these findings suggest that cyclic stretch may be altering the intracellular transport of plasmids to increase gene expression. PMID- 12727120 TI - Development of a formulation that enhances gene expression and efficacy following intraperitoneal administration in rabbits and mice. AB - We conducted a series of experiments to determine if intraperitoneal (IP) delivery of recombinant adenovirus (rAd)-based therapies is improved through carrier vehicle selection, and compared an icodextrin solution (a high molecular weight dextrin with a prolonged peritoneal cavity residence time) with a standardized phosphate buffered saline (PBS) delivery solution. In vitro, comparative adenovirus particle concentration determination (27 h) and bioactivity assay (24h) indicated equivalent compatibility with icodextrin or PBS. In vivo, rabbits treated IP (100 ml) with rAd-betagal 1 x 10(9) P/ml in icodextrin showed improved transgene expression throughout the peritoneal wall compared to rAd-betagal in PBS. In PC-3 tumor-bearing mice treated IP with 5 x 10(9) P/0.5 ml or 1 x 10(10) P/0.5 ml rAd-betagal, transgene expression was significantly enhanced (p < 0.01) with icodextrin compared to PBS in both tumor specimens and peritoneal wall. In subsequent studies we compared prolongation of survival in intraperitoneal PC-3 and MDAH-2774 human xenograft tumor models in nude mice using rAd-p53 in icodextrin or PBS in multi-dose ranging (1 x 10(8) to 1 x 10(10) P) experiments. The icodextrin formulation alone significantly increased rAd-p53 mediated survival (p < 0.05). In animals, these results show that IP rAd gene therapy can be improved with the use of icodextrin, and suggest that prolonged retention and distribution in the peritoneal cavity is an important factor. PMID- 12727119 TI - Targeted delivery of DNA to the mitochondrial compartment via import sequence conjugated peptide nucleic acid. AB - We report that oligonucleotides can be introduced into the mitochondria of living mammalian cells by annealing them to peptide nucleic acids coupled to mitochondrial targeting peptides. These complexes are imported into the mitochondrial matrix through the outer and inner membrane import channels of isolated mitochondria. They are also imported into the mitochondria of cultured cells, provided that the cytosolic uptake of the complexes is facilitated by using synthetic polycations or membrane permeabilizing toxins. Our method now promises to provide a viable strategy for the genetic modification of the mitochondria in cultured cells, animals and patients. PMID- 12727121 TI - Developmental antecedents of anxiety sensitivity. AB - Anxiety sensitivity (AS) has been implicated in the etiology of anxiety and depressive disorders. Nonetheless, little is known about the developmental antecedents of AS or about its role as a mediator of emotional distress. To investigate these issues, 249 university students completed a battery of self report measures. Regression analyses and partial correlations were used to evaluate hypotheses. Analyses revealed that exposure to parental threatening, hostile, and rejecting behaviors (1) predicted overall AS and (2) appeared differentially related to AS factors. Additionally, AS acted as a mediator between parenting and both current emotional distress and history of emotional disorder symptoms. These data support the notion that parental behaviors hypothesized as etiologic in the development of emotional disorders may also be etiologic in the development of AS and that AS may serve as a mediator between childhood experience and emotional distress. PMID- 12727122 TI - Threat perception abnormalities in children: the role of anxiety disorders symptoms, chronic anxiety, and state anxiety. AB - This study investigated the relative contribution of general (trait) anxiety and state anxiety to threat perception abnormalities in nonreferred children aged 8 13 years (N=299). Children were first asked to complete self-report measures of anxiety disorders symptoms and chronic anxiety. Next, they were individually interviewed using an ambiguous story paradigm from which a number of threat perception indexes were derived. Just before the interview started, children were asked to fill out a measure of state anxiety. Results showed that high levels of general anxiety (as indexed by anxiety disorders symptoms and chronic anxiety) were significantly related to increased threat perception and lower threat thresholds. High levels of state anxiety were also associated with increased threat perception and lower threat thresholds. Regression analyses indicated that general anxiety and state anxiety both accounted for a unique proportion of the variance in threat perception abnormalities, although the contribution of general anxiety was in most cases substantially larger than that of state anxiety. Finally, no support was found for the notion that threat perception abnormalities are the result of the conjoint influence of general anxiety and state anxiety. PMID- 12727123 TI - The relationship between dispositional pessimistic attributional style versus trauma-specific attributions and PTSD symptoms. AB - Because a relatively small percentage of individuals who experience a traumatic event ultimately develop posttraumatic stress disorder (PTSD), it is incumbent upon researchers to identify factors of vulnerability and risk. One possible risk factor is attributional style or the types of causes individuals habitually offer for negative life events. This study examined the association between pessimistic attributional style and symptoms of PTSD. Because of methodological problems with the traditional questionnaire measurement of dispositional attributional style, this investigation added a structured content analysis of participants' trauma narratives to examine associations between trauma-specific attributions and PTSD symptoms. Dispositional attributional style, measured by the attributional style questionnaire (ASQ), was significantly associated with PTSD symptoms, but trauma specific attributions more strongly predicted symptoms. PMID- 12727124 TI - Further investigation of the obsessive-compulsive inventory: psychometric analysis in two non-clinical samples. AB - The obsessive-compulsive inventory [OCI; Psychol. Assessment 10 (1998) 206-214] is a self-report measure of obsessive-compulsive disorder (OCD) intended for use with both clinical and non-clinical samples (). Two reports support its convergence with established measures of OCD and its assessment of specific symptom subtypes (e.g., checking and washing). The current studies investigate the OCI factor structure in two non-clinical samples. Based on correlational analyses and complementary factor analyses, Study 1 results indicate that five subscales, rather than the seven rationally derived ones, best capture the structure of symptoms measured by the OCI. These findings were replicated in Study 2. Based on these analyses, we offer suggestions for revising the original subscales into these five empirically derived subscales that closely resemble structures proposed in the current OCD literature. PMID- 12727125 TI - Panic control treatment for agoraphobia. AB - The goal of the present study was to compare the efficacy of cognitive-behavioral treatment for panic control alone versus this treatment containing an additional in vivo exposure component. The sample was comprised of 68 individuals who met diagnosis for panic disorder with agoraphobia. Participants were randomly assigned to one of two 16-week treatment conditions, panic control only and panic control with in vivo exposure. Assessments were repeated at baseline, mid treatment, posttreatment, and 6-month follow-up using diagnostic and behavioral measures. Results indicated that the two treatment conditions were equally efficacious for both panic disorder and agoraphobia. The intervention explicitly targeting agoraphobia appeared superfluous given the efficacy of panic control alone. On the other hand, reduction in panic frequency predicted reduction in agoraphobic avoidance overall. The practical and theoretical implications are discussed, as are limitations and directions for future research. PMID- 12727126 TI - Fear responses to mock magnetic resonance imaging among college students: toward a prototype experiment. AB - Two hundred randomly selected student participants (139 females, 61 males) responded initially to questionnaires that quantified variables such as state and trait anxiety, anxiety sensitivity, claustrophobia, and panic/agoraphobia. Later they were informed that a mock magnetic resonance imaging (MRI) procedure was upcoming, and were prompted to provide self-efficacy ratings vis-a-vis completing the procedure. Finally, the participants' behavioral reactions to a mock MRI procedure were characterized; their heart beats were recorded and ratings of fearfulness were acquired. One purpose of the research was simply to tally numbers of participants who responded fearfully in various ways: 7 failed the procedure behaviorally, 7 others completed the procedure but did so fearfully, 17 others completed the procedure but manifested excessive heart-rate responsivity. A second purpose of the research was to "predict" subjects' fear-response categorization psychometrically and/or with self-efficacy ratings: psychometric data related to claustrophobia predicted subjects' fear-response categorization as did self-efficacy ratings. According to these results mock MRI assessment among college students provides a promising context for research on claustrophobia. PMID- 12727127 TI - Theoretical and empirical exploration of the similarities between emotional numbing in posttraumatic stress disorder and alexithymia. AB - This study rationally and scientifically examines the similarities between the emotional numbing symptoms of posttraumatic stress disorder (PTSD) and alexithymia. A clinical database from the Iowa City Department of Veterans Affairs Medical Center consisting of data from 274 combat veterans was used. Significant positive correlations were found between measures of PTSD, alexithymia, and combat exposure. Principal components analysis supported a lack of independence between PTSD subscales and alexithymia, as all variables loaded on one factor best labeled PTSD. This article presents both theoretical and empirical evidence suggesting that in traumatized persons, alexithymia may be better conceptualized as the emotional numbing aspect of PTSD rather than as a distinct construct. PMID- 12727128 TI - Use of a Symptom Validity Test in the forensic assessment of Posttraumatic Stress Disorder. AB - Symptom Validity Tests (SVTs) employ a two-alternative, forced-choice method to assess an individual's motivation to perform. The strategy is routinely used by neuropsychologist with plaintiffs who allege brain injury, but heretofore has not been applied to the forensic assessment of posttrauma claims. The present case report applied an accepted SVT, the Portland Digit Recognition Test, to the forensic assessment of Posttraumatic Stress Disorder. Findings illustrate how SVTs can be useful in any forensic or disability case that presents with memory complaints. PMID- 12727129 TI - Social stress differentially regulates neuroendocrine responses in laying hens: I. Genetic basis of dopamine responses under three different social conditions. AB - Effects of genetic-environmental interactions on plasma dopamine (DA) concentrations were studied in White Leghorn chickens selected for both high (HGPS) or low (LGPS) group productivity and survivability resulting from cannibalism and flightiness. Plasma DA levels were measured from chickens in three social treatments: single-, two-, or ten-hen cages. The two-hen treatment consisted of paired chickens from three genetic lines: HGPS, LGPS and a commercial strain, Dekalb XL (DXL). In HGPS/DXL and LGPS/DXL pairs, the DXL hen was used as a standardized genetic competitor. The ten-hen treatment contained only hens from the same line, which is similar to the original selection condition. After 7 weeks housing in the social environments, LGPS hens in the ten hen treatment had greater plasma DA concentrations than HGPS hens (P<0.05). Compared to levels in the ten-hen treatment from the same line, plasma DA concentrations in both HGPS and LGPS hens were significantly lower in the two-hen treatment (average mean, 0.09 vs. 0.15 ng/ml and 0.22 vs. 0.44 ng/ml, P<0.05, respectively), but significantly higher in the single-hen treatment (average mean, 0.44 vs. 0.15 ng/ml and 1.78 vs. 0.44 ng/ml, P<0.05 and P<0.01, respectively). In the single-hen treatment, LGPS hens had greater plasma DA levels than HGPS hens (P<0.05). The results provide evidence of genetically related differences in the regulation of chickens' plasma DA concentrations in response to social stress. These differences may magnify the behavioral and physiological differences observed in the lines under basal and challenged conditions. These results suggest that these chicken lines may provide a new model for investigating effects of DA on the control of behavioral, neural and endocrine responses to stress. PMID- 12727131 TI - Dopamine and serotonin function in untreated schizophrenia: clinical correlates of the apomorphine and d-fenfluramine tests. AB - This study examined the prolactin (PRL), adrenocorticotropin (ACTH) and cortisol responses to the direct DA receptor agonist apomorphine (APO) and the selective 5HT-releasing agent d-fenfluramine (d-FEN) in 20 untreated inpatients with DSM-IV schizophrenia and without a history of suicide attempt, compared to 23 hospitalized healthy controls. We hypothesized that different patterns of responsiveness of the DA and 5-HT systems might be associated with specific schizophrenic symptom clusters. A positive correlation was observed between pituitary-adrenal response to APO and d-FEN tests (i.e. deltaACTH and deltacortisol) in the overall population and in schizophrenic patients. Pituitary adrenal response to APO was lower in patients than in normal controls. Moreover, lower pituitary-adrenal response to APO and d-FEN was associated with increased severity of BPRS thought disturbance score. Lower pituitary-adrenal responses to APO (and to a lesser degree to d-FEN) differentiated paranoid from disorganized schizophrenic patients. Neither PRL suppression to APO, nor PRL stimulation to d FEN were altered in schizophrenic patients. Our results suggest that decreased hypothalamic DA receptor activity (possibly secondary to increased presynaptic DA release) together with relatively decreased 5-HT tone characterize paranoid SCH, while normal hypothalamic DA receptor activity together with relatively increased 5-HT tone characterize the disorganized SCH subtype. PMID- 12727130 TI - Quality of life of growth hormone (GH) deficient young adults during discontinuation and restart of GH therapy. AB - The present study evaluates the effects of one year of discontinuation and one year of growth hormone (GH) treatment on quality of life (QoL) in young adults with childhood-onset growth hormone deficiency (CO-GHD). Twenty-two subjects (14 males, 8 females; 11 isolated growth hormone deficient [IGHD], 11 multiple pituitary hormone deficient [MPHD]), aged between 15 and 22 years, on ongoing GH treatment were assessed during one year of discontinuation. Thereafter, 9 of these patients, who were found to be still GH deficient (GHD), added by 11 newly recruited GHD patients who also were not treated in the preceding year (in total 10 males and 10 females, aged between 17 and 27, 5 IGHD, 15 MPHD), restarted GH treatment for one year. During discontinuation and restart of GH treatment somatic and psychological assessments took place every 6 months. In the first 6 months of the GH discontinuation period insulin-like growth factor I (IGF-I) level significantly declined whereas no further decrease in IGF-I was seen after month 6. The number of psychological complaints and depression increased only during the first 6 months of discontinuation. Across the 12-month of discontinuation tension increased in MPHD and decreased in IGHD patients. Only in the first 6 months of GH treatment IGF-I level increased, anxiety decreased and QoL improved. Depression scores tended to decrease across the 12 month treatment period. During the 2-year discontinuation and treatment period intra-subject IGF I level was negatively correlated with depression, fatigue, tension and anxiety and positively with vigor and memory. At the end of the treatment period all psychometric parameters were similar or even improved compared to those at the start of the discontinuation period. It is concluded that one year discontinuation of GH treatment leads to a decrease in QoL within 6 months which effect is counteracted within 6 months after restart of GH treatment. PMID- 12727132 TI - HIV-positive men differ in immunologic but not catecholamine response to an acute psychological stressor. AB - Acute psychological stress in humans induces sudden alterations in catecholamine plasma levels and in the distribution of peripheral blood lymphocytes. The purpose of this study was to investigate whether infection with the human immunodeficiency virus (HIV) had an impact on the psychoneuroimmunologic axis. Twelve asymptomatic HIV-positive homo- or bisexual men (CD4 cell counts>400/mm3) and 13 healthy HIV-negative control subjects were exposed to an acute psychological stressor consisting of a 30 min semi-structured stress interview using emotion- and client-centered conversation techniques surrounded by two relaxation periods. Changes in neuroendocrine and immunological, as well as cardiovascular parameters, were intermittently monitored. Under the influence of the stressor plasma norepinephrine (NE) levels increased significantly in HIV positive patients (+30.6%; p<0.05) and in HIV-negative individuals (+17.5%; n.s.). These changes were paralleled by significant increases in blood pressure and heart rate. Plasma cortisol decreased continuously from initially high levels during the entire experimental setting in both groups, compatible with an adaption reaction. Concomitantly, WBC and neutrophilic granulocytes increased significantly in HIV-negative subjects, while they were blunted in HIV-positive patients. Interestingly, NK cell increases were significantly higher during the stress experiment in HIV-positive patients than in HIV-negative controls. CD4+ and B cell counts remained unaffected by the stressor. These results suggest that catecholamine secretion induced by an acute psychological stressor is preserved in HIV-infected patients with the responsiveness of WBC and neutrophilic granulocytes being diminished, while NK cells showed an increased response. PMID- 12727133 TI - Olanzapine's effects to reduce fear and anxiety and enhance social interactions coincide with increased progestin concentrations of ovariectomized rats. AB - Administration of olanzapine, an antipsychotic drug, can dose-dependently increase the levels of progesterone's metabolite, 5 alpha-pregnane-3 alpha-ol-20 one (3 alpha,5 alpha-THP) in the brain, which may have anxiolytic effects. The purpose of this experiment was to investigate the effects of olanzapine administration on anxiety behavior and progestin levels. Ovariectomized (ovx) rats (N=33) were administered olanzapine (i.p.: 5.0 or 10.0 mg/kg) or vehicle (saline buffered with acetic acid) and an hour later were tested for motor and anxiety behavior (n=8 per group) or had tissue collected for measurement of progestin concentrations (n=3 per group). Rats that were administered 5.0 or 10.0 mg/kg of olanzapine spent less time freezing in response to shock in the defensive burying task, spent more time on the open arms of the elevated plus maze, and spent more time in social interaction with a conspecific than did vehicle-administered rats. Olanzapine (5.0 or 10.0 mg/kg, i.p.) significantly increased plasma and produced non-significant increases in whole brain concentrations of progesterone and 3 alpha,5 alpha-THP compared to that seen following vehicle administration. Together, these data are consistent with the hypothesis that olanzapine reduces fear, has anxiolytic effects, and may enhance social interaction in part due to increasing progestin concentrations. PMID- 12727134 TI - Impact of a unilateral brain lesion on cortisol secretion and emotional state: anterior/posterior dissociation in humans. AB - The main goal of this study was to evaluate whether a unilateral brain lesion in a human population is associated with a modification of the circadian cortisol secretion profile, and/or patient's emotional state. The second goal of this study was to assess whether there would be differences in both the pattern of cortisol secretion and emotional state in brain-damaged patients as a function of side of lesion, and localization (anterior vs posterior) of lesion. Eight patients with a left cortical lesion, six patients with a right cortical lesion, four patients with basal ganglia lesions (2 left and 2 right) and ten healthy volunteers were evaluated daily on measures of salivary cortisol levels and subjective feelings of joy and sadness at 0700, 1200, 1600 and 1900 hours over a 15-day period. Patients with cortical brain lesions presented higher cortisol levels and higher scores of sadness at the time of the morning peak (7:00 am), when compared to healthy volunteers and patients with basal ganglia lesions. Laterality of the lesion was not related to cortisol secretion, but frontal damage (anterior lesion) was associated with higher cortisol levels at the time of the morning peak (7:00 am) when compared to more posterior damage. There was no significant correlation between basal circulating levels of cortisol and emotional states in patients and healthy subjects. The results of this study suggest that hypothalamic-pituitary-adrenal (HPA) axis dysregulation is associated with unilateral injury particularly in frontal areas. These results, obtained in a human population, go along with recent animal studies reporting an implication of frontal regions in HPA activity. PMID- 12727135 TI - Characterization and functional significance of glucocorticoid receptors in patients with major depression: modulation by antidepressant treatment. AB - Hyperactivity of the hypothalamic pituitary adrenal (HPA) axis in patients with major depression is one of the most consistent findings in biological psychiatry. Experimental data support the idea that glucocorticoid-mediated feedback via glucocorticoid receptors (GR) is impaired in major depression. The aim of the present work was to assess the putative changes in GR density of peripheral blood mononuclear cells (PBMCs) in a group of patients with major depression and to determine modulation of these GR sites by antidepressant treatment. In addition, susceptibility of PBMCs to glucocorticoid effects was also studied using a functional end-point analysis in vitro, such as cortisol inhibition of mitogen induced lymphocyte proliferation. Cortisol levels were also measured before and after dexamethasone suppression test (DST). The results showed a decrease in GR density in depressed patients compared with healthy subjects, mainly in those patients that showed basal cortisol levels in the upper normal range and were refractory to DST. Regarding the functional significance of this variation, two representative groups emerged from our study: a) free-medication patients with GR function comparable to healthy controls, and b) patients showing diminished GR activity. These results suggest a lack of relationship between GR density and cortisol-induced inhibition of lymphocyte proliferation. Patients treated with different antidepressant drugs showed a marked increase in the number of GR sites per cell compared to non-treated. Interestingly, this increase was even higher than in normal subjects. Hence, restoration of GR density after an efficient antidepressant treatment could be an index of an effective modulatory action of drugs on GR expression and highlights the possibility that GR levels might be used as markers of a successful treatment. PMID- 12727137 TI - CTLA-4, T helper lymphocytes and dendritic cells: an internal perspective of T cell homeostasis. AB - In mice and humans, the size of the peripheral lymphocyte pool remains relatively constant throughout adult life, in the absence of disease. Among the factors that influence the survival and homeostasis of T cells, external stimuli such as infections have long been considered to be of primary importance. However, emerging data indicate that internal stimuli, including self-peptide (presented in association with major-histocompatibility-complex molecules) and cytokines, might also control the size and composition of T-cell pools. Recent evidence suggests that cytotoxic-T-lymphocyte-associated antigen 4, an immune attenuator, contributes significantly to the homeostatic control of T-helper-cell proliferation through mechanisms that are both intrinsic and extrinsic to the T cell. PMID- 12727136 TI - Voluntary wheel running modulates glutamate receptor subunit gene expression and stress hormone release in Lewis rats. AB - Lewis rats that are known to be addiction-prone, develop compulsive running if they have access to running wheels. The present experiments were aimed 1) to evaluate the activation of stress systems following chronic and acute voluntary wheel running in Lewis rats by measurement of hormone release and gene expression of neuropeptides related to hypothalamic-pituitary-adrenocortical (HPA) axis activity and 2) to test the hypothesis that wheel running as a combined model of addictive behavior and stress exposure is associated with modulation of ionotropic glutamate receptor subunits in the ventral tegmental area. Voluntary running for three weeks but not for one night resulted in a rise in plasma corticosterone and adrenocorticotropic hormone (ACTH) levels (p<0.05) compared to those in control rats. Principal component analysis revealed the relation between POMC gene expression in the intermediate pituitary and running rate. Acute exposure of animals to voluntary wheel running induced a significant decrease in alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptor GluR1 subunit mRNA levels (p<0.01), while repeated voluntary physical activity increased levels of GluR1 mRNA in the ventral tegmentum (p<0.05). Neither acute nor chronic wheel running influenced N-methyl-D-aspartate (NMDA) receptor subunit NR1 mRNA levels in the ventral tegmental area. Thus, the present study revealed changes in AMPA receptor subunit gene expression in a reward-related brain structure as well as an activation of HPA axis in response to compulsive wheel running in Lewis rats. It may be suggested that hormones of HPA axis and glutamate receptors belong to the factors that substantiate higher vulnerability to addictive behavior. PMID- 12727138 TI - Genetic associations: false or true? AB - Genetic association studies for multigenetic diseases are like fishing for the truth in a sea of trillions of candidate analyses. Red herrings are unavoidably common, and bias might cause serious misconceptions. However, a sizeable proportion of identified genetic associations are probably true. Meta-analysis, a rigorous, comprehensive, quantitative synthesis of all the available data, might help us to separate the true from the false. PMID- 12727139 TI - Could TCR antagonism explain associations between MHC genes and disease? AB - Alleles of major histocompatibility complex (MHC) loci are associated with certain types of diseases, including those of infectious and autoimmune origin. MHC products can promote susceptibility or resistance to disease by stimulating or inhibiting immune responses. Recent evidence suggests that MHC-associated peptides derived from self-proteins can act as antagonists of T-cell activation, thereby inhibiting immune responses to antigens. We suggest that self-peptide promoted antagonism might explain some associations between MHC alleles and particular chronic diseases. PMID- 12727140 TI - Hydrolytic nucleoside and nucleotide deamination, and genetic instability: a possible link between RNA-editing enzymes and cancer? AB - Post-transcriptional RNA editing generates novel gene products by changing the coding sequence of the transcript from that in the genome. Two classes of RNA editing exist in mammals, each of which involves an enzymatic deamination. These reactions have stringent sequence and structural requirements for their target RNAs, and each requires distinctive enzymatic machinery. Alterations in the expression or abundance of RNA-editing factors produce unanticipated alterations in the processing or expression of RNAs, in some cases outside their physiological targets. Recent findings suggest that unregulated expression of the cytidine-deaminase gene family might lead to deamination of deoxycytidine nucleotides in DNA. Aberrant or dysregulated RNA editing, or altered expression of editing factors, might contribute to genomic instability in cancer. PMID- 12727141 TI - A fatal attraction: Mycobacterium tuberculosis and HIV-1 target DC-SIGN to escape immune surveillance. AB - Dendritic cells (DCs) are vital in the defense against pathogens. However, it is becoming increasingly clear that some pathogens subvert DC functions to escape immune surveillance. For example, HIV-1 targets the DC-specific C-type lectin DC SIGN (DC-specific intercellular-adhesion-molecule-3-grabbing nonintegrin) to hijack DCs for viral dissemination. Binding to DC-SIGN protects HIV-1 from antigen processing and facilitates its transport to lymphoid tissues, where DC SIGN promotes HIV-1 infection of T cells. Recent studies demonstrate that DC-SIGN is a universal pathogen receptor that also recognizes Ebola, cytomegalovirus and mycobacteria. Mycobacterium tuberculosis targets DC-SIGN by a mechanism that is distinct from that of HIV-1, leading to inhibition of the immunostimulatory function of DC and, hence, promotion of pathogen survival. A better understanding of DC-SIGN-pathogen interactions and their effects on DC function should help to combat infections. PMID- 12727142 TI - S-nitrosylation in health and disease. AB - S-nitrosylation is a ubiquitous redox-related modification of cysteine thiol by nitric oxide (NO), which transduces NO bioactivity. Accumulating evidence suggests that the products of S-nitrosylation, S-nitrosothiols (SNOs), play key roles in human health and disease. In this review, we focus on the reaction mechanisms underlying the biological responses mediated by SNOs. We emphasize reactions that can be identified with complex (patho)physiological responses, and that best rationalize the observed increase or decrease in specific classes of SNOs across a spectrum of disease states. Thus, changes in the levels of various SNOs depend on specific defects in both enzymatic and non-enzymatic mechanisms of nitrosothiol formation, processing and degradation. An understanding of these mechanisms is crucial for the development of an integrated model of NO biology, and for effective treatment of diseases associated with dysregulation of NO homeostasis. PMID- 12727143 TI - Protein carbonylation in human diseases. AB - Oxidative modifications of enzymes and structural proteins play a significant role in the aetiology and/or progression of several human diseases. Protein carbonyl content is the most general and well-used biomarker of severe oxidative protein damage. Human diseases associated with protein carbonylation include Alzheimer's disease, chronic lung disease, chronic renal failure, diabetes and sepsis. Rapid recent progress in the identification of carbonylated proteins should provide new diagnostic (possibly pre-symptomatic) biomarkers for oxidative damage, and yield basic information to aid the establishment an efficacious antioxidant therapy. PMID- 12727144 TI - Apoptosis in heart: basic mechanisms and implications in cardiovascular diseases. AB - Apoptosis has been implicated in both acute and chronic heart diseases. The loss of myocardium is an important pathogenic process in heart and, hence, the inhibition of apoptosis is a promising therapeutic option. Significant progress has been made in demonstrating the role of apoptosis in various heart diseases, and in elucidating the molecular mechanisms of cardiac apoptosis. In addition, the effects of various hypertrophic signaling molecules on apoptosis have been characterized. However, the significance of apoptosis to the overall pathogenesis of heart failure, and the relationship between cardiac hypertrophic signaling and apoptosis still needs further investigation. PMID- 12727145 TI - Right ventricular tachycardia-arrhythmogenic right ventricular cardiomyopathy or idiopathic? PMID- 12727146 TI - Noninvasive evaluation of ischaemic heart disease: myocardial perfusion imaging or stress echocardiography? PMID- 12727147 TI - Clinical and electrophysiological differences between patients with arrhythmogenic right ventricular dysplasia and right ventricular outflow tract tachycardia. AB - AIMS: Radiofrequency catheter ablation is considered first line treatment for symptomatic patients with right ventricular outflow tract tachycardia (RVOT). The role of ablation in arrhythmogenic right ventricular dysplasia (ARVD) is more limited. As such, differentiating between the two conditions is essential. METHODS AND RESULTS: This study compared non-invasive findings, magnetic resonance images (MRI), invasive electrophysiological characteristics, results of ablation and long-term outcome in 50 consecutive patients with RVOT (33) or ARVD (17). Structural abnormalities were uniform in the ARVD group; in addition 18 (54%) of the RVOT tachycardia group had MRI abnormalities. At electrophysiological study the tachycardia in the ARVD group displayed features of re-entry in over 80%, but behaved with a triggered automatic basis in 97% with RVOT. Ablation was complete or partial success in 12 (71%) patients with ARVD and ventricular tachycardia (VT) recurred in eight (48%). In the RVOT patients, ablation was a complete success in 97% with recurrent VT in 6%. Long-term success in the RVOT patients was 95% in both patients with and without MRI abnormalities. CONCLUSIONS: Electrophysiological characterization can differentiate ARVD from RVOT. The finding of abnormalities on MRI does not have any bearing on arrhythmia mechanism, acute or long-term success of RFA. PMID- 12727148 TI - Development and prospective validation of a risk stratification system for patients with syncope in the emergency department: the OESIL risk score. AB - AIMS: Aim of the present study was the development and the subsequent validation of a simple risk classification system for patients presenting with syncope to the emergency departments. METHODS AND RESULTS: A group of 270 consecutive patients (145 females, mean age 59.5 years) presenting with syncope to the emergency departments of six community hospitals of the Lazio region of Italy was used as a derivation cohort for the development of the risk classification system. Data from the baseline clinical history, physical examination and electrocardiogram were used to identify independent predictors of total mortality within the first 12 months after the initial evaluation. Multivariate analysis allowed the recognition of the following predictors of mortality: (1) age >65 years; (2) cardiovascular disease in clinical history; (3) syncope without prodromes; and (4) abnormal electrocardiogram. The OESIL (Osservatorio Epidemiologico sulla Sincope nel Lazio) score was calculated by the simple arithmetic sum of the number of predictors present in every single patient. Mortality increased significantly as the score increased in the derivation cohort (0% for a score of 0, 0.8% for 1 point; 19.6% for 2 points; 34.7% for 3 points; 57.1% for 4 points; p<0,0001 for trend). A similar pattern of increasing mortality with increasing score was prospectively confirmed in a second validation cohort of 328 consecutive patients (178 females; mean age, 57.5 years). CONCLUSIONS: Clinical and electrocardiographic data available at presentation to the emergency department can be used for the risk stratification of patients with syncope. The OESIL risk score may represent a simple prognostication tool that could be usefully employed for the triage and management of patients with syncope in emergency departments. PMID- 12727149 TI - Association of a genetic variant of endothelial nitric oxide synthase with the 1 year clinical outcome after coronary stent placement. AB - AIMS: Endothelial nitric oxide synthase (eNOS) catalyzes the formation of nitric oxide which has vasodilatory, antithrombotic, antiinflammatory and antiproliferative properties. The TT genotype of the single nucleotide polymorphism 894 G/T, located in exon 7 of the eNOS gene, was found to be associated with coronary spasm, coronary artery disease (CAD) and myocardial infarction (MI). We investigated the possibility that the 894 TT genotype has an unfavorable impact on the angiographic and clinical outcome after the placement of stents in coronary arteries. METHODS AND RESULTS: Our study included 1850 patients with CAD who were treated with stent implantation. Major adverse clinical events, including death, MI, and target vessel revascularization, were recorded for 1 year after the intervention. Patients with genotype 894 TT had an increase in the risk of death or MI (hazard ratio 2.14, 95% confidence interval (CI) 1.23-3.72; p=0.007), if compared with G allele carriers. TT patients showed no significant increase in the risk for angiographic restenosis (odds ratio (OR) 1.11, 95% CI 0.78-1.56; p=0.56) and target vessel revascularization (OR 1.21, 95% CI 0.82-1.78; p=0.34). CONCLUSIONS: In comparison with eNOS 894 G allele carriers, patients of the TT genotype were at an increased risk of death or MI within 1 year after coronary artery stenting. PMID- 12727150 TI - Early revascularization is associated with improved survival in elderly patients with acute myocardial infarction complicated by cardiogenic shock: a report from the SHOCK Trial Registry. AB - AIMS: The SHould we emergently revascularize Occluded Coronaries in cardiogenic shocK (SHOCK) Trial showed no benefit of early revascularization in patients aged >/=75 years with acute myocardial infarction and cardiogenic shock. We examined the effect of age on treatment and outcomes of patients with cardiogenic shock in the SHOCK Trial Registry. METHODS AND RESULTS: We compared clinical and treatment factors in patients in the SHOCK Trial Registry with shock due to pump failure aged <75 years (n=588) and >/=75 years (n=277), and 30-day mortality of patients treated with early revascularization <18 hours since onset of shock and those undergoing a later or no revascularization procedure. After excluding early deaths covariate-adjusted relative risk and 95% confidence intervals were calculated to compare the revascularization strategies within the two age groups. Older patients more often had prior myocardial infarction, congestive heart failure, renal insufficiency, other comorbidities, and severe coronary anatomy. In-hospital mortality in the early vs. late or no revascularization groups was 45 vs. 61% for patients aged <75 years (p=0.002) and 48 vs. 81% for those aged >/=75 years (p=0.0003). After exclusion of 65 early deaths and covariate adjustment, the relative risk was 0.76 (0.59, 0.99; p=0.045) in patients aged <75 years and 0.46 (0.28, 0.75; p=0.002) in patients aged >/=75 years. CONCLUSIONS: Elderly patients with myocardial infarction complicated by cardiogenic shock are less likely to be treated with invasive therapies than younger patients with shock. Covariate-adjusted modeling reveals that elderly patients selected for early revascularization have a lower mortality rate than those receiving a revascularization procedure later or never. PMID- 12727151 TI - Under utilisation of evidence-based treatment partially explains for the unfavourable prognosis in diabetic patients with acute myocardial infarction. AB - AIMS: The prognosis after an acute myocardial infarction is worse for patients with diabetes mellitus than for those without. We investigated whether differences in the use of evidence-based treatment may contribute to the differences in 1-year survival in a large cohort of consecutive acute myocardial infarction patients with and without diabetes mellitus. METHODS: We included patients below the age of 80 years from the Register of Information and Knowledge about Swedish Heart Intensive care Admissions (RIKS-HIA), which included all patients admitted to coronary care units at 58 hospitals during 1995-1998. In all 5193 patients had the combination of acute myocardial infarction and diabetes mellitus while 20440 had myocardial infarction but no diabetes diagnosed. Multivariate logistical regression analyses were performed to evaluate the influence of diabetes mellitus on the use of evidence-based treatment and its association with survival during the first year after the index hospitalisation. RESULTS: The prevalence of diabetes mellitus was 20.3% (males 18.5%; females 24.4%). The 1-year mortality was substantially higher among diabetic patients compared with those without diabetes mellitus (13.0 vs. 22.3% for males and 14.4 vs. 26.1% for female patients, respectively) with an odds ratio (OR) (95% confidence interval (CI)) in three different age groups: <65 years 2.65 (2.23 3.16); 65-74 years 1.81 (1.61-2.04) and >75 years 1.71 (1.50-1.93). During hospital stay patients with diabetes mellitus received significantly less treatment with heparins (37 vs. 43%; p<0.001), intravenous beta blockade (29 vs. 33%; p<0.001), thrombolysis (31 vs. 41%; p<0.001) and acute revascularisation (4 vs. 5%; p<0.003). A similar pattern was apparent at hospital discharge. After multiple adjustments for dissimilarities in baseline characteristics between the two groups, patients with diabetes were significantly less likely to be treated with reperfusion therapy (OR 0.83), heparins (OR 0.88), statins (OR 0.88) or to be revascularised within 14 days from hospital discharge procedures (OR 0.86) while the use of ACE-inhibitors was more prevalent among diabetic patients compared to non-diabetic patients (OR 1.45). The mortality reducing effects of evidence-based treatment like reperfusion, heparins, aspirin, beta-blockers, lipid-lowering treatment and revascularisation were, in multivariate analyses, of equal benefit in diabetic and non-diabetic patients. INTERPRETATION: Diabetes mellitus continues to be a major independent predictor of 1-year mortality following an acute myocardial infarction, especially in younger age groups. This may partly be explained by less use of evidence-based treatment although treatment benefits are similar in both patients with and without diabetes mellitus. Thus a more extensive use of established treatment has a potential to improve the poor prognosis among patients with acute myocardial infarction and diabetes mellitus. PMID- 12727152 TI - Prognostic value of extensive coronary calcium quantities in symptomatic males--a 5-year follow-up study. AB - AIMS: Coronary calcium scores (CSs) have been shown to predict future events in patients presenting for first-time evaluation of CAD. Long-term outcome data on symptomatic subjects with advanced CAD are limited. In this study, we evaluated the prognostic value of very high coronary CSs in symptomatic males undergoing angiography and analyzed the impact of event definitions on identification of risk predictor's. METHODS AND RESULTS: Fifty consecutive symptomatic males with electron beam computed tomography (EBT)-based CSs >1000 were matched 1:2 by age with symptomatic males with scores between 400-1000 and 100-400. All 150 patients underwent coronary angiography. CAD risk factors were ascertained. Events were analyzed after 5 years for: (1) hard coronary events (coronary death and myocardial infarction); (2) overall hard events (adding stroke and non-coronary deaths); and (3) all events (including long-term revascularizations). During follow-up, 17 deaths, two infarctions and three strokes occurred in 21 patients; 38 patients underwent 43 revascularizations. Events occurred earlier and more frequently in patients with scores >1000. Left main disease was the only independent predictor of hard coronary events (hazard ratio, 4.5; 95% confidence interval, 1.1-17.8). Left main disease (4.3; 1.4-13.0) and CSs (1.7; 1.1-2.5) independently predicted overall hard events. Only CSs>90th percentile independently predicted all events (2.5; 1.3-4.8). CONCLUSIONS: Symptomatic males with extensive CSs carry an even higher risk for future events than other symptomatic males with advanced CAD. In these patients, EBT-based calcified plaque burden and angiographic indices of disease severity may have a complementary role in predicting future cardiovascular events. PMID- 12727153 TI - Does rheumatic myocarditis really exists? Systematic study with echocardiography and cardiac troponin I blood levels. AB - AIMS: Revised guidelines for diagnosis of rheumatic fever indicate that rheumatic myocarditis may 'contribute' to the genesis of congestive heart failure. Our objective was to assess non-invasively the presence of non-clinical markers of myocardial involvement in acute rheumatic fever. METHODS: Echocardiography and assessment of cardiac troponin I (cTnI) blood levels were systematically performed in 95 consecutive patients with acute rheumatic fever, who were divided into three groups. Group 1: patients without carditis (n=22); group 2: patients with carditis and without congestive heart failure (n=59); group 3: patients with carditis and congestive heart failure (n=14). RESULTS: Left ventricular ejection fraction was normal in all patients and did not differ between groups (group 1: 0.72+/-0.08, group 2: 0.69+/-0.06, and group 3: 0.66+/-0.07, p=0.09). Left ventricular diameters tend to be larger in group 3, but all patients had severe mitral and/or aortic regurgitation. Mean cTnI was 0.077+/-0.017 ng/ml (normal <0.1 ng/ml), did not differ between groups (p=0.45), and only 13 patients (seven with pericardial effusion) had detectable levels (0.2-0.4 ng/ml). CONCLUSIONS: Our study neither detected cTnI elevations nor echocardiographic abnormalities suggesting significant myocardial involvement during rheumatic fever. Congestive heart failure was always associated to severe valve regurgitation. PMID- 12727154 TI - Long-term survival in patients hospitalized with congestive heart failure: relation to preserved and reduced left ventricular systolic function. AB - AIMS: The purpose of this study was to evaluate the influence of left ventricular systolic function on the survival in a large consecutive cohort of patients hospitalized with congestive heart failure and to determine how left ventricular systolic function interacts with co-morbid conditions in terms of prognosis. METHODS AND RESULTS: Analysis of survival data from 5491 patients admitted for new or worsening heart failure to 34 departments of cardiology or internal medicine in Denmark from 1993-1996 was carried out. A standardized echocardiogram was available for 95% of the patients, and left ventricular systolic function was estimated using wall motion index score. Follow-up time was 5-8 years. Patients with preserved systolic function were older, more frequently female, and had less evidence of ischemia than patients with systolic dysfunction. After 1 year, 24% of the patients had died. Low wall motion index was a potent independent predictor of death (risk ratio for one unit increase, 0.60 (0.56-0.64)), and was of greater prognostic significance in younger patients and patients with a history of myocardial ischemia. However, even in patients with preserved systolic function, mortality was high (1 year mortality, 19%). CONCLUSION: In hospitalized heart failure patients, particularly in younger patients with ischemic heart disease, mortality risk is inversely related to left ventricular systolic function. PMID- 12727155 TI - A randomised study of home-based electrical stimulation of the legs and conventional bicycle exercise training for patients with chronic heart failure. AB - AIMS: Recent guidelines recommend regular exercise in the management of patients with chronic heart failure (CHF). This study was designed to compare the safety and efficacy of conventional bicycle exercise and functional electrical stimulation (FES) of the legs as forms of home-based exercise training for patients with stable CHF. METHODS AND RESULTS: Forty-six patients (38 male) with stable NYHA Class II/III heart failure underwent a 6-week training programme using either a bicycle ergometer or electrical stimulation of the quadriceps and gastrocnemius muscles. In the bike group, significant increases were seen in 6 min walk (44.6m, 95% confidence interval (CI) 29.3-60.9 m), treadmill exercise time (110 s, 95% CI 72.2-148.0 s), maximum leg strength (5.32 kg, 95% CI 3.18 7.45 kg), and quadriceps fatigue index (0.08, 95% CI 0.04-0.12) following training. In the stimulator group, similar significant increases were seen following training for 6-min walk (40.6m, 95% CI 28.2-53.0m), treadmill exercise time (67 s, 95% CI 11.8-121.8s), maximum leg strength (5.35 kg, 95% CI 1.53-9.17 kg), and quadriceps fatigue index (0.10, 95% CI 0.04-0.17). Peak VO(2)did not change in either group following training, indicating a low-intensity regime. Quality of life scores improved following training when the bicycle and stimulator groups were considered together, but not when considered separately ( 0.43, 95% CI -8.13 to -0.56). CONCLUSIONS: FES produces beneficial changes in muscle performance and exercise capacity in patients with CHF. Within this study, the benefits were similar to those observed following bicycle training. FES could be offered to patients with heart failure as an alternative to bicycle training as part of a home-based rehabilitation programme. PMID- 12727156 TI - Cardiac resynchronization in chronic heart failure: some considerations about the cost-effectiveness. PMID- 12727158 TI - Is prophylaxis the best use of the ICD? Response to the letter to the editor (Sharma et al.). PMID- 12727159 TI - Chlamydia pneumoniae IgA titres and coronary heart disease: prospective study and meta-analysis. PMID- 12727160 TI - The logistic EuroSCORE. PMID- 12727161 TI - Complications of tracheal and thoracic surgery: the role of multisection helical CT and computerized reformations. AB - Helical computed tomography (CT) has an important role in the evaluation of a wide range of congenital and acquired thoracic abnormalities. The development of advanced computerized reformations enables the generation of bronchographic and bronchoscopic images of the tracheobronchial tree, as well as angiographic images of pulmonary arteries and veins. Additionally, it provides coronal and sagittal reconstruction imaging of parenchymal abnormalities. This information is obtained by a 20-30s procedure on a typical single channel system, which makes helical CT an optimal technique for the evaluation of patients undergoing major upper airways and thoracic interventions. The recent introduction of multisection CT scanners allows faster imaging of patients with thinner collimation, thus improving spatial resolution along the longitudinal (z) axis of the patient along with reduction of motion artefacts. This article demonstrates the use of dual and quad-section helical CT in the postoperative evaluation of patients undergoing laryngo-tracheal and thoracic interventions, including laryngoplasty, tracheal endoscopic laser ablation, lobectomy, pneumonectomy, lung transplantation, sleeve resection, pulmonary angioplasty, and pulmonary artery thromboendarterectomy. Emphasis is given to the additive value of using computerized reformations over axial images, especially for delineation of complex postoperative anatomical details in the tracheobronchial tree and pulmonary vasculature. PMID- 12727162 TI - Sonography of neck lymph nodes. Part I: normal lymph nodes. AB - Grey scale and power Doppler sonography play an important role in assessment of cervical lymphadenopathy. However, before examination of pathological nodes, a clear understanding of the anatomy of cervical nodes, scanning technique and sonographic appearances of normal cervical nodes is essential. This article reviews these topics in order to provide a baseline for sonographic examination of cervical lymphadenopathy. PMID- 12727163 TI - Sonography of neck lymph nodes. Part II: abnormal lymph nodes. AB - Assessment of cervical lymph nodes is essential for patients with head and neck carcinomas, and ultrasound is a useful imaging technique. Sonographic features that help distinguish between the causes of neck lymphadenopathy, including grey scale and Doppler features, are discussed. In addition to the distribution and location of nodes, the useful grey-scale features are: size, shape, internal architecture, intranodal necrosis, absence of hilar structure and calcification. The useful Doppler features are: distribution of vascularity and intranodal resistance. Ancillary features such as oedema of soft tissue and nodal matting are particularly helpful to identify tuberculous nodes. PMID- 12727164 TI - The role of ultrasound-guided cytology of groin lymph nodes in the management of squamous cell carcinoma of the vulva: 5-year experience in 44 patients. AB - AIM: To assess the accuracy of ultrasound combined with fine-needle aspiration cytology (FNAC) in the detection of lymph node metastasis in patients with squamous cell carcinoma of the vulva. MATERIALS AND METHODS: The groin nodes of 44 consecutive patients with primary squamous cell carcinoma of the vulva undergoing groin node dissection were assessed with ultrasound and FNAC. The results were compared with histology from subsequent inguinofemoral lymph node dissection. Twenty-nine patients underwent bilateral groin node dissections and 15 unilateral providing comparable data for 73 groins. RESULTS: Histology demonstrated metastatic disease in 28 groins and no evidence of metastatic disease in 45. Ultrasound agreed with the histology in 67 of the 73 groins (92%), with two false-positives, four false-negatives and two indeterminate appearances. Cytology agreed with the histology in 65 of 72 FNAC samples obtained (90%), with six false-negatives, and one indeterminate result. No false-positive cytology results were seen. Ultrasound and FNAC together failed to detect metastatic disease in four groins, one with an indeterminate ultrasound appearance, another with indeterminate cytology, the two others each having a single positive inguinal node despite a negative ultrasound and FNAC. CONCLUSION: The combination of ultrasound and FNAC provides a sensitive and specific tool for pre-operative assessment and may prevent unnecessary groin dissection and the attendant morbidity in selected patients with vulval cancer. PMID- 12727165 TI - Ultrasound detection of abdominal lymph nodes in chronic liver diseases. A retrospective analysis. AB - AIM: To retrospectively evaluate the prevalence of lymph nodes of the hepato duodenal ligament in a group of patients with chronic liver disease of various aetiologies and to investigate what clinical, aetiological and laboratory data may lead to their appearance. MATERIALS AND METHODS: One thousand and three patients (554 men, 449 women) were studied, including 557 with chronic hepatitis and 446 with liver cirrhosis. The presence of lymph nodes near the trunk of the portal vein, hepatic artery, celiac axis, superior mesenteric vein and pancreas head was investigated using ultrasound. RESULTS: Lymph nodes were detected in 394 out of the 1003 study patients (39.3%); their number ranged from one to four, with a diameter ranging between 0.8 and 4 cm. The highest prevalence was in the subgroup of patients with primary biliary cirrhosis (87.5%), followed by patients with hepatitis C virus (HCV; 42%), patients with HCV and hepatitis B virus (HBV; 41.3%), autoimmune hepatitis (40%), and HBV alone (21.2%). In the alcoholic and idiopathic subgroups prevalence was 9.5%, while in the non-alcoholic steatohepatitis and haemochromatosis subgroups it was 0%. HCV RNA was present in 97 out of 103 lymph node-positive patients and in 141 out of 168 lymph node negative HCV-negative patients (p<0.003). Lymphadenopathy frequency increased as the liver disease worsened (chi(2) MH=74.3; p<0.0001). CONCLUSION: Despite the limitations of a retrospective study, our data indicate a high prevalence of lymphadenopathy in liver disease patients; ultrasound evidence of lymph nodes of the hepato-duodenal ligament in a given liver disease may most likely suggest a HCV or an autoimmune aetiology and a more severe histological picture. PMID- 12727166 TI - Multislice computed tomographic coronary angiography: experience in a UK centre. AB - AIM: To evaluate the technique of coronary angiography with retrospectively electrocardiogram (ECG)-gated four-slice helical computed tomography (CT). MATERIALS AND METHODS: Within 1 month of undergoing routine day-case diagnostic coronary angiography, 30 consecutive patients also underwent retrospectively ECG gated multislice CT coronary angiography. This enabled direct comparison of seven segments of proximal and mid-coronary artery for each patient by two blinded assessors. Each segment of coronary artery from the multislice CT image was evaluated initially for "assessability" and those segments deemed assessable were subsequently investigated for the presence or absence of a significantly (n=70%) stenotic lesion. RESULTS: Overall 68% of proximal and mid-coronary artery segments were assessable. The sensitivity and specificity of four-slice CT coronary angiography in assessable segments for detecting the presence or absence (n=70%) of stenoses were 72 and 86%, respectively. These results correspond to a positive predictive value of 53% and a 93% negative predictive value. If the 32% of non-assessable segments are added into the calculation then the sensitivity and specificity fall to 49 and 66%, respectively. CONCLUSION: Although multislice CT coronary angiography is a promising technique, the overall assessability and diagnostic accuracy of four-slice CT acquisition is not sufficient to justify routine clinical use. Further, evaluation should investigate the benefit of the reduction in temporal and spatial resolution offered by 16 and 32 slice acquisition. PMID- 12727167 TI - Magic angle imaging of the achilles tendon in patients with chronic tendonopathy. AB - AIMS: To assess the Achilles tendon in patients with chronic tendonopathy using magnetic resonance (MR) magic angle imaging, and to compare the appearances and uptake of contrast medium in abnormal tendons with those in normal tendons. MATERIAL AND METHODS: Eight patients with chronic Achilles tendonopathy and five normal controls were examined with the long axis of the tendon placed at 55 degrees and at 0 degrees to the main magnetic field. Conventional two-dimensional (2D) multi-slice images were obtained and T1 values were calculated before, and for up to 1h after the administration of intravenous gadodiamide. Both the unenhanced appearance and the pattern of enhancement in the tendon were compared. RESULTS: In the patients with tendonopathy, high signal intensity areas were evident on the short T1 inversion recovery (STIR) images obtained at 55 degrees in all tendons. Contrast medium enhancement was seen in six tendons and was most obvious on the images obtained at the magic angle. This was initially focal and then spread more diffusely within the tendon. After contrast medium administration, T1 values were significantly reduced in the tendonopathy group compared with normal controls (p<0.01). On the late post-contrast medium images obtained at 55 degrees, enhancement was evident in most of the tendon and correlated well with high signal intensity seen on STIR images. CONCLUSION: The use of magic angle MR imaging improved the demonstration of signal changes in the Achilles tendon in chronic tendonopathy. The STIR images obtained at the magic angle showed more obvious signal change than those obtained at 0 degrees. The changes due to enhancement were much more evident on images obtained at 55 degrees than at 0 degrees. The uptake of contrast medium was greater in the patients than in normal controls. PMID- 12727168 TI - Diurnal T2 value changes in the lumbar intervertebral discs. AB - AIM: To investigate if there is any in vivo diurnal T2 value changes in the nucleus pulposus in normal and degenerated lumbar intervertebral discs. MATERIALS AND METHODS: Eighteen volunteers (16 men, two women) with no symptoms related to lumbar disease were imaged in the morning before beginning daily work and in the evening after finishing work. Sagittal fast spin-echo T2-weighted images were obtained, following spin-echo transverse imaging for T2 analysis. RESULTS: The normal disc group (n=71) showed statistically significant diurnal T2 value changes (p<0.001), whilst the degenerating disc group (n=19) showed no significant diurnal changes. The normal disc group up to age 34 years showed statistically significant diurnal T2 value changes. However no significant changes were demonstrated after the age 35 years. CONCLUSION: Statistically significant diurnal T2 value changes in the normal lumbar intervertebral discs were demonstrated. Disappearance of the diurnal T2 value changes in the normal discs after the age 35 years was revealed for the first time and thought to be an aspect of aging, not caused by degeneration. PMID- 12727169 TI - Small bowel obstruction--the water-soluble follow-through revisited. AB - AIMS: The aim of the present study to examine the use of a modified water-soluble follow-through in the diagnosis and management of small bowel obstruction (SBO). MATERIALS AND METHODS: Sixty-two patients were recruited to the study: 33 into the control group and 29 into the study group. A modified small bowel follow through (SBFT) was performed in the study group patients. The control group was managed conventionally. Assessment was made by questionnaire documenting initial surgical diagnosis and likelihood of operative intervention, final diagnosis and surgical outcome (operative versus non-operative). RESULTS: SBFT changed the diagnosis in 12/24 of the study group (p<0.01). In the study group 8/24 proceeded to surgery whereas 19/33 underwent laparotomy in the control group, representing a relative risk reduction of 52%, but this was not statistically significant (0.10>p>0.05, chi-squared test). CONCLUSION: SBFT remains a valid and useful tool in surgical management of SBO. In particular it aids diagnostic confidence in planning surgical intervention, particularly in uncomplicated patients. PMID- 12727170 TI - Gastrostomy insertion: comparing the options--PEG, RIG or PIG? AB - AIM: To compare percutaneous endoscopic gastrostomy (PEG) with radiologically inserted gastrostomy (RIG) and assess a hybrid gastrostomy technique (per-oral image-guided gastrostomy, PIG). MATERIALS AND METHODS: Fifty PEGs and 50 RIGs performed in three centres were prospectively compared and the endoscopic findings of 200 PEGs reviewed. A fluoroscopy-guided technique was modified to place 20 F over-the-wire PEG-tubes in 60 consecutive patients. RESULTS: Technical success was 98%, 100% and 100% for PEG, RIG and PIG, respectively. Antibiotic prophylaxis significantly reduced stoma infection for orally placed tubes (p=0.02). Ten out of 50 (20%) small-bore RIG tubes blocked. Replacement tubes were required in six out of 50 PEGs (12%), 10 out of 50 RIGs (20%), but no PIGs (p<0.001). No procedure-related complications occurred. The function of radiologically placed tubes was significantly improved with the larger PIG (p<0.001), with similar wound infection rates. PIG was successful in 24 patients where endoscopic insertion could not be performed. Significant endoscopic abnormalities were found in 42 out of 200 PEG patients (21%), all related to peptic disease. Insignificant pathology was found in 8.5%. CONCLUSION: PIG combines advantages of both traditional methods with a higher success and lower re-intervention rate. Endoscopy is unlikely to detect clinically relevant pathology other than peptic disease. PIG is a very effective gastrostomy method; it has better long-term results than RIG and is successful where conventional PEG has failed. PMID- 12727171 TI - Mechanical small bowel obstruction due to intraluminal thrombus in a patient with acute leukaemia: ultrasonic findings. PMID- 12727172 TI - Identification of inguinal lymph node metastases from vulval carcinoma by magnetic resonance imaging: an initial report. PMID- 12727174 TI - How should we deliver an out of hours nephrostomy service? PMID- 12727179 TI - Semantic versus phonological false recognition in aging and Alzheimer's disease. AB - Patients with Alzheimer's disease (AD) have been found to exhibit lower levels of false recognition of semantic associates compared with healthy older adults. Because these patients may show impaired performance of episodic and semantic memory tasks, this finding could be explained by deficits in episodic memory, semantic memory, or both. The authors adapted a paradigm for comparison of semantic versus phonological false recognition. They found that: (a) patients with AD exhibited lower levels of corrected false recognition of semantic, phonological, and hybrid (mixed semantic and phonological) lists than older adults, and (b) patients with AD showed very similar levels of false recognition for all list types. These results suggest that only episodic memory deficits are necessary to explain the lower level of false recognition of semantic associates observed in patients with AD when compared to older adults. Additionally, (c) older adults showed greater levels of semantic, phonological, and hybrid false recognition than younger adults, extending previous false recognition research of semantically related words and categorized colored photographs to phonologically related words. PMID- 12727180 TI - Machiavellianism and Theory of Mind in people affected by schizophrenia. AB - CONTEXT: People affected by schizophrenia show deficits in social interaction (Cramer, Bowen, & O'Neill, 1992); such a deficit may stem from an alteration in the representation of mental states. OBJECT: In this study we have examined 42 people affected by schizophrenia, diagnosed according to DSM-IV criteria, by testing them with an instrument (the Mach IV scale, Christie & Geis, 1970) assessing "Machiavellian Intelligence" according to Niccolo Machiavelli's assertions. Schizophrenic subjects were compared to a sample of normal controls, matched for age, sex, and education. OBJECTIVE: The aim of our study was to evaluate if there were correlations between the ability to understand other people's mental states and the capacity for interpreting "Machiavellian expressions." RESULTS: Results show that people affected by schizophrenia are less able to use Machiavellian-style "tactical" abilities than controls. CONCLUSION: Such a disability seems to be correlated to poor mentalising abilities. PMID- 12727182 TI - Differential brain responses when applying criterion attribute versus family resemblance rule learning. AB - Subsystems of category learning have been identified on the basis of general domains of content (e.g., tools, faces). The present study examined categories from the standpoint of internal structure and determined brain topography associated with expressing two fundamentally different category rule structures (criterion attribute, CA, and family resemblance, FR). CA category learning involves processing stimuli by isolated features and classifying by properties held by all members. FR learning involves processing stimuli by integral wholes and classifying on overall similarity among members without sharing identical features. fMRI BOLD response to CA and FR categorization was measured with pseudowords as stimuli. Category knowledge for both tasks was mastered prior to brain imaging. Areas of activation emerged unique to the structure of each category and followed from the nature of the rule abstraction procedure. CA categorization was implemented by strong target monitoring and expectation (medial parietal), rule maintenance in working memory, feature selection processes (inferior frontal), and a sensitivity to high frequency components of the stimulus such as isolated features (anterior temporal). FR categorization, consistent with its multi-featural nature, involved word-level processing (left extrastriate) that evoked articulatory rehearsal (medial cerebellar). The data suggest category structure is an important determinant of brain response during categorization. For instance, anterior temporal structures may help attune visual processing systems to high frequency components to support the learning of criterial, highly predictive rules. PMID- 12727181 TI - Perceptual, affective, and cognitive judgments of odors: pleasantness and handedness effects. AB - The present study sought to examine the differential processing of pleasant, neutral, and unpleasant odors. The effects of the nostril stimulated (left or right) and the type of judgment (perceptual, affective, or cognitive) performed on the olfactory stimuli were also studied. To this end, 64 subjects were asked to smell pleasant, neutral, and unpleasant odors under four conditions (detection, intensity, pleasantness, and familiarity tasks). The participants were to perform these tasks as quickly as possible, while response times were recorded. The results showed that (i) unpleasant odors were assessed more rapidly than neutral or pleasant odors, and that this was specifically true (ii) during right nostril stimulation, and (iii) during pleasantness assessment, suggesting possible differential cerebral hemisphere involvement, with a right-side advantage for processing of unpleasant affect in olfaction. A handedness effect on familiarity judgment is also discussed. PMID- 12727183 TI - Generating an image from an ambiguous visual input: an electroencephalographic (EEG) investigation. AB - Fourteen right-handed males were shown slides of common objects (e.g., wristwatch), familiar situations (e.g., two people shaking hands), and inkblots from a popular projective test (Holtzman, 1986) and then asked to name the object, assess the situation, or describe the most salient image emerging from the inkblot. Alpha power suppression was monitored over the left and right frontal, temporal, parietal, and occipital lobes to determine the brain circuitry responsible for the processing of each type of stimulus. When processing common objects and familiar situations there was bilateral activation of the parietal and occipital lobes; when processing inkblots, bilateral activation of the parietal and occipital lobes was again obtained, but complemented by selective activation of the right frontal lobe. The later suggests that anterior regions of the right cerebral hemisphere contribute to the generation of dynamic images like those evoked by visually ambiguous inkblots. PMID- 12727184 TI - Phased mirror tracing outcomes correlate with several hemisphericity measures. AB - Recently, subjects, separated into two large groups by the Dichotic Deafness Test (DDT), a new brain laterality measure. These were significantly correlated with groups separated by two preference-based hemisphericity-type questionnaires, the Preference Questionnaire (PrefQ) and Polarity Questionnaire (PolQ). Here, Mirror Tracing, another potential biophysical measure of hemisphericity earlier reported to be weakly correlated with laterality, was tested using 171 subjects, the DDT, and both questionnaires. No correlations between the DDT, PrefQ, and PolQ separation methods and Mirror Tracing outcomes were found until individuals identified as having left brain affect were removed from the population. Then, robust correlations appeared for the remaining right-affect subjects alone (n=82). These correlations became even higher when the left brain affect subjects that had been removed (n=89) were "phase corrected" from a contralateral to an ipsilateral connectivity for motor dominance and then returned to the sample (n=171). These Phased Mirror-Tracing (PMT) outcomes were also significantly correlated with 10 of the 11 statements of the PolQ. Thus, PMT joins the DDT as a second performance-based measure of laterality whose outcomes significantly correlated with those of preference-based hemisphericity questionnaires. PMID- 12727185 TI - Two-hand line-bisection task outcomes correlate with several measures of hemisphericity. AB - While seeking new functional methods to reassess the concept of hemisphericity, a two-hand line-bisection task was investigated because of reports of large, stable differences among the general population. These were found to be due to hemispheric differences in judgment of the midpoint of horizontal lines, made visible due to the unilateral brain control of each hand. By use of a two-hand line-bisection test (Best-Hand Test), university workers (n=412) were readily sorted into theoretical response categories, resulting in the production of two large groups. These two groups correlated well with those produced by four independent hemisphericity assessments, two physiological and two psychological. This is the third biophysical method whose performance-based group separations significantly correlated with those produced by preference-based hemisphericity type questionnaires. It is rapid and avoids language, education, or cultural bias. PMID- 12727186 TI - Asymmetric influences of temporally vs. nasally presented masked visual information: evidence for collicular contributions to nonconscious priming effects. AB - It was tested whether the retino-collicular projection allows for the processing of nonconsciously registered visual information in healthy individuals. Masked primes were presented to different visual hemifields. Because the retino collicular projection is stronger for temporal than for nasal hemifields, priming should be stronger by temporal than by nasal primes. This pattern was tested in two experiments (Experiments 1 and 3). Further, with less peripheral primes, only available to weaker parts of the retino-collicular projection, hemifield asymmetries of priming vanished (Experiment 2). In conclusion, the study offers first evidence for collicular contributions to nonconscious priming effects by visual information in healthy individuals. PMID- 12727187 TI - Learning to control brain activity: a review of the production and control of EEG components for driving brain-computer interface (BCI) systems. AB - Brain-computer interface (BCI) technology relies on the ability of individuals to voluntarily and reliably produce changes in their electroencephalographic (EEG) activity. The present paper reviews research on cognitive tasks and other methods of generating and controlling specific changes in EEG activity that can be used to drive BCI systems. To date, motor imagery has been the most commonly used task. This paper explores the possibility that other cognitive tasks, including those used in imaging studies, may prove to be more effective. Other factors which influence performance are also considered in relation to selection of tasks, as well as training of subjects. PMID- 12727188 TI - Rapid eye movement sleep dreaming is characterized by uncoupled EEG activity between frontal and perceptual cortical regions. AB - EEG coherent activity is involved in the binding of spatially separated but temporally correlated stimuli into whole events. Cognitive features of rapid eye movement sleep (REM) dreaming resemble frontal lobe dysfunction. Therefore, temporal coupling of EEG activity between frontal and perceptual regions was analyzed from 10 min prior to dream reports (8 adults) from stage-2 and REM sleep. EEG correlation between frontal and perceptual regions decreased and, among perceptual regions increased during REM. The temporal dissociation of EEG activity between executive and perceptual regions supplies an inadequate mechanism for the binding and interpretation of ongoing perceptual activity resulting in dream bizarreness. PMID- 12727189 TI - Children with developmental coordination disorder (DCD) and their ability to disengage ongoing attentional focus: more on inhibitory function. AB - Children with developmental coordination disorder (DCD), along with their Control counterparts, completed two endogenous, spatial precue tasks. When the precue arrow was informative (.80) with respect to target location, the spatial precue effect results demonstrated that children with DCD take significantly longer than Control individuals to volitionally disengage (inhibit) attention from an endogenously cued location (i.e., a disengagement inhibition deficit). When the precue was uninformative (.25), we found, contrary to a common assumption, that the precue arrow automatically moved attention in the direction of the arrow, and, in addition, that DCD children may also be less able to inhibit the precued induced urge to move attention (i.e., an initiation inhibition deficit). This type of inhibitory difficulty was also indicated for manual response inclinations produced on catch trials. Overall, DCD children appeared to have an elevated difficulty suppressing the initiation of incorrect, stimulus-provoked movement urges, be they manual or attention in nature. PMID- 12727190 TI - The self-composing brain: towards a glial-neuronal brain theory. AB - A brain model is proposed which describes its structural organization and the related functions as compartments organized in time and space. On a molecular level the negative feedback loops of clock-controlled genes are interpreted as compartments. This spatio-temporal operational principle may also work on the cellular level as glial-neuronal interactions, wherein glia have a spatio temporal boundary setting function. The synchronization of the multi compartmental operations of the brain is compared to the harmonization in a symphony and appears as an integrated behavior of the whole organism, defined as modes of behavior. For explanation of the principle of harmonization, an example from Schubert's Symphony No. 8 has been chosen. While harmonization refers to the synchronization of diverse systems, it seems appropriate to select the brain of a composer and the structure of musical composition as a paradigm towards a glial neuronal brain theory. Finally, some limitations of experimental brain research are discussed and robotics are proposed as a promising alternative. PMID- 12727191 TI - Dissociation between the mental rotation of visual images and motor images in unilateral brain-damaged patients. AB - Deficits in the mental rotation of body parts and of external objects can be doubly dissociated (Rumiati, Tomasino, Vorano, Umilta, & De Luce, 2001; Sirigu & Duhamel, 2001; Tomasino, Rumiati, & Umita, in press). The aim of this study was to replicate this finding and to then investigate the relevance of the specific hemispheres in these deficits. Nine patients with unilateral lesions (five in the Left Hemisphere and four in the Right) and 20 control subjects, performed a single task requiring mental rotation of hands, and two tasks requiring mental rotation of external objects. RH patients were impaired in the rotation of external objects, but showed intact performance on the rotation of hands; the opposite pattern was found for LH patients. These results support the view that the LH contributes to the mental rotation of hands, recruiting processes specific to motor preparation, while the RH is specialized for mental rotation of external objects. PMID- 12727192 TI - Asymmetry questionnaire outcomes correlate with several hemisphericity measures. AB - The asymmetry questionnaire segregated subjects (n=143) into two groups. These were significantly correlated with similar groups separated, not only by three new biophysical hemisphericity protocols (Dichotic Deafness Test, Phased Mirror Tracing, Best Hand Test), but also by two preference-type measures (polarity questionnaire, preference questionnaire). Each of the 15 asymmetry questionnaire statements was significantly correlated with the outcomes of these five laterality measures. This is the third questionnaire whose outcomes correlate with those of the new biophysical measures of hemisphericity. PMID- 12727193 TI - Familial handedness and spatial ability: a study with Chinese students aged 14 24. AB - This study tested Annett's right-shift theory on spatial ability with two samples from China. The Vandenberg Mental Rotation Test (MRT), Edinburgh Handedness Inventory, and Family Handedness Questionnaire were administered to 266 high school students and 297 undergraduates. We found very few r++ or r-- among Chinese students. Most Chinese are either moderately right-handed or ambidextrous. Consistent with Casey's finding, we found using different methods to classify handedness leads to different conclusions. However, we did not find the effect of familial handedness that Casey found. Visual strategy is related to success on the MRT but handedness is not. PMID- 12727194 TI - Expression of Na+/HCO3- cotransporter and its role in pH regulation in mouse parotid acinar cells. AB - Ion transporters such as Na(+)/H(+) exchanger (NHE), Cl(-)/HCO(3)(-) exchanger (AE), and Na(+)/HCO(3)(-) cotransporter (NBC) are known to contribute to the intracellular pH (pH(i)) regulation during agonist-induced stimulation. This study examined the mechanisms for the pH(i) regulation in the mouse parotid and sublingual acinar cells using the fluorescent pH-sensitive probe, BCECF. The pH(i) recovery from agonist-induced acidification in the sublingual acinar cells was completely blocked by EIPA, a NHE inhibitor. However, the parotid acinar cells required DIDS, a NBC1 inhibitor, in addition to EIPA in order to block the pH(i) recovery. Moreover, RT-PCR analysis detected the expression of pancreatic NBC1 (pNBC1) only in the parotid acinar cells. These results provide strong evidence that the mechanisms for the pH(i) regulation are different in the two types of acinar cells, and pNBC1 contributes to pH(i) regulation in the parotid acinar cells, whereas NHE is likely to be the exclusive pH(i) regulator in the sublingual acinar cells. PMID- 12727195 TI - The cargo receptor ERGIC-53 is a target of the unfolded protein response. AB - The accumulation of unfolded proteins in the ER triggers a signaling response known as unfolded protein response (UPR). In yeast the UPR affects several hundred genes that encode ER chaperones and proteins operating at later stages of secretion. In mammalian cells the UPR appears to be more limited to chaperones of the ER and genes assumed to be important after cell recovery from ER stress that are not important for secretion. Here, we report that the mRNA of lectin ERGIC 53, a cargo receptor for the transport of glycoproteins from ER to ERGIC, and of its related protein VIP36 is induced by the known inducers of ER stress, tunicamycin and thapsigargin. In parallel, the rate of synthesis of the ERGIC-53 protein was induced by these agents. The response was due to the UPR since it was also triggered by castanospermine, a specific inducer of UPR, and inhibited by genistein. Thapsigargin-induced upregulation of ERGIC-53 could be fully accounted for by the ATF6 pathway of UPR. The results suggest that in mammalian cells the UPR also affects traffic from and beyond the ER. PMID- 12727196 TI - Properties of purified recombinant human polyamine oxidase, PAOh1/SMO. AB - The discovery of an inducible oxidase whose apparent substrate preference is spermine indicates that polyamine catabolism is more complex than that originally proposed. To facilitate the study of this enzyme, the purification and characterization of the recombinant human PAOh1/SMO polyamine oxidase are reported. Purified PAOh1/SMO oxidizes both spermine (K(m)=1.6 microM) and N(1) acetylspermine (K(m)=51 microM), but does not oxidize spermidine. The purified human enzyme also does not oxidize eight representative antitumor polyamine analogues; however, specific oligamine analogues were found to be potent inhibitors of the oxidation of spermine by PAOh1/SMO. The results of these studies are consistent with the hypothesis that PAOh1/SMO represents a new addition to the polyamine metabolic pathway that may represent a new target for antineoplastic drug development. PMID- 12727197 TI - Read-through histone transcripts containing 3' adenylate tails are zygotically expressed in Xenopus embryos and undergo processing to mature transcripts when introduced into oocyte nuclei. AB - Messages encoding replication-dependent histone genes generally terminate with a stem-loop structure and lack polyadenylate tails. Adenylated histone transcripts were identified in Xenopus oocytes, though the role of the adenylate tracts is unknown. We report isolation of cDNAs from Xenopus embryos encoding histone mRNAs with 3' adenylate tracts. They also contain targets for stem-loop binding protein and U7 snRNA, which are required for histone RNA processing. One sequence is a read-through transcript containing a complete version of the downstream gene from the anti-parallel strand, similar to the RNAs from lampbrush loops of Notophthalmus oocytes. We injected read-through transcripts into Xenopus oocyte nuclei and they were processed to mature histone RNAs. Our results suggest that addition of 3' adenylate sequences might be a normal part of histone RNA synthesis. Also, these results shed light on the enigma of the developmental regulation of adenylated histone transcripts in Xenopus oocytes. PMID- 12727198 TI - Method to overcome photoreaction, a serious drawback to the use of dichlorofluorescin in evaluation of reactive oxygen species. AB - Non-fluorescent dichlorofluorescin (DCFH) was converted to fluorescent products by photo-irradiation during observations with spectrofluorometer and fluorescence microscopy. Photo-irradiation of DCFH at 250, 300, 330, 400, 500, or 600 nm generated fluorescent dichlorofluorescein (DCF), an oxidation product of DCFH, and an unrecognized fluorescent product. The ratio of the unknown product to DCF varied from 0.15 to 8.21 depending on wavelength. Although reactive oxygen species scavengers, such as catalase, superoxide dismutase, and sodium azide, did not suppress the increase in non-specified fluorescence, reagents such as ascorbic acid, mercaptopropionyl glycine, and methoxycinnamic acid, in a cell free system, almost completely suppressed it with little effect on the fluorescence of DCF. Meanwhile, ascorbic acid also suppressed non-specified fluorescence in cells, but not completely. At low concentrations of DCFH, the speed of increasing fluorescence was considerably retarded, to such a degree that the fluorescence increase in cells during fluorescence microscopic observation was negligible. The addition, at the time of evaluation, of the above reagents to cell-free systems and, in cell systems, reducing the concentration of DCFH, effectively suppressed the photoreaction of DCFH. PMID- 12727199 TI - Engineering nuclear localization signals in modular protein vehicles for gene therapy. AB - Amino acids from 126 to 135 of the SV40 virus T antigen act as efficient nuclear localization signal during infection but also when fused to recombinant proteins. This peptide has been inserted into two alternative acceptor sites of a modified Escherichia coli beta-galactosidase which also displays a DNA-binding domain, a cell-binding motif for integrin alpha(v)beta(3) targeting and cell internalization, and a cryptic nuclear targeting signal naturally present in the bacterial enzyme. In cultured cells, the presence of the SV40 peptide enhances the expression of a delivered DNA up to 30-fold. However, the DNA expression levels are largely depending on the chosen insertion site for the SV40 segment concomitant to the structural impact of peptide accommodation on the protein vehicle. The structural stability of the hybrid protein, apparently critical for efficient gene transfer, is discussed in the context of modular protein engineering to develop non-viral vectors for gene therapy. PMID- 12727201 TI - DNA damage by ethylbenzenehydroperoxide formed from carcinogenic ethylbenzene by sunlight irradiation. AB - Ethylbenzene, widely used in human life, is a non-mutagenic carcinogen. Sunlight irradiated ethylbenzene caused DNA damage in the presence of Cu2+, but unirradiated ethylbenzene did not. A Cu+ -specific chelator bathocuproine inhibited DNA damage and catalase showed a little inhibitory effect. The scopoletin assay revealed that peroxides and H(2)O(2) were formed in ethylbenzene exposed to sunlight. These results suggest that Cu+ and alkoxyl radical mainly participate in DNA damage, and H(2)O(2) partially does. When catalase was added, DNA damage at thymine and cytosine was inhibited. Ethylbenzenehydroperoxide, identified by GC/MS analysis, induced the formation of 8-oxo-7,8-dihydro-2' deoxyguanosine and caused DNA damage at consecutive guanines, as observed with cumenehydroperoxide. Equimolar concentrations of H(2)O(2) and acetophenone were produced by the sunlight-irradiation of 1-phenylethanol, a further degraded product of ethylbenzene. These results indicate a novel pathway that oxidative DNA damage induced by the peroxide and H(2)O(2) derived from sunlight-irradiated ethylbenzene may lead to expression of the carcinogenicity. PMID- 12727200 TI - Vitamin D-dependent recruitment of DNA-PK to the chromatinized negative vitamin D response element in the PTHrP gene is required for gene repression by vitamin D. AB - The mechanism of transcriptional repression by nuclear hormone receptors, especially in the presence of the ligands, is largely unknown. We previously reported that 1,25-dihydroxyvitamin D(3) (1,25 vitamin D3) inhibited expression of the parathyroid hormone-related polypeptide (PTHrP) gene through the interaction between the liganded monomeric vitamin D receptor (VDR) and the negative DNA element in the PTHrP gene (nVDRE(RP)). In this study, we employed chromatin immunoprecipitation (ChIP) assay and confirmed that 1,25 vitamin D3 recruited DNA-dependent protein kinase (DNA-PKcs) to the chromatinized nVDRE(RP). Conversely, the regulatory subunits of DNA-PK were associated with the nVDRE(RP) sequences only when 1,25 vitamin D3 was absent. VDR was constitutively associated with these chromatinized nVDRE(RP) sequences. Furthermore, DNA-PKcs could phosphorylate VDR in vitro. We raise a possibility that a conformational change of VDR through its phosphorylation mediated by DNA-PKcs underlies the mechanism of gene repression by 1,25 vitamin D3-bound VDR. PMID- 12727202 TI - Effect of transforming growth factor-beta on calcium homeostasis in prostate carcinoma cells. AB - Ca(2+) plays a fundamental role in the control of a variety of cellular functions, in particular, in energy metabolism and apoptosis. In this study, we show that TGF-beta at concentrations of 0.1-1.0 ng/ml transiently increases the level of intracellular Ca(2+) ([Ca(2+)](in)) in human prostate carcinoma, PC-3U, cells. Experiments with mitochondrial inhibitors (oligomycin and antimycin A) and an inhibitor of endoplasmic reticulum Ca(2+) uptake (BHQ) implied that the effect of TGF-beta1 was due to an effect on the mitochondria. TGF-beta1 treatment resulted in a decrease in ATP synthesis and to a depolarisation, leading to a release of Ca(2+) from mitochondria and decreased activity of the Ca(2+) pumps. Analysis of the mitochondria within the PC-3U cells by polarography and membrane potential-sensitive dye (Rhodamine 123) confirmed that under these experimental conditions, TGF-beta1 inhibited ATP synthesis and depolarised the mitochondria. The results implicate that TGF-beta1 affects the function of the mitochondria and may be of significance for the understanding of the proapoptotic effect of TGF beta1 in these cells. PMID- 12727203 TI - Contribution of hydrogen peroxide to the cytotoxicity of green tea and red wines. AB - Green tea and red wine are claimed to have health benefits because of their high content of polyphenolic compounds, but they have also been reported as mutagenic in some test systems. In this paper, we show that a commonly used cell culture medium, Dulbecco's modified Eagle's medium (DMEM), catalyses oxidation of green tea and red wines to generate H(2)O(2). The level of H(2)O(2) produced from green tea accounted for all of the cytotoxic effects of this beverage on PCl2 cells. By contrast, H(2)O(2) was only responsible for part of the cytotoxicity of the red wines examined. Our data illustrate the danger of extrapolating from cell culture studies to predict the effects of complex beverages in vivo. PMID- 12727204 TI - Ca2+/calmodulin binds and dissociates K-RasB from membrane. AB - We have investigated the interaction of calmodulin (CaM) with Ras-p21 and the significance of this association. All Ras-p21 isoforms tested (H-, K-, and N-Ras) were detected in the particulate fraction of human platelets and MCF-7 cells, a human breast cancer cell line. In MCF-7 cells, H- and N-Ras were also detected in the cytosolic fraction. K-RasB from platelet and MCF-7 cell lysates was found to bind CaM in a Ca2+ -dependent but GTPgammaS-independent manner. The yeast two hybrid analysis demonstrated that K-RasB binds to CaM in vivo. Incubation of isolated membranes from platelet and MCF-7 cells with CaM caused dissociation of only K-RasB from membranes in a Ca2+ -dependent manner. CaM antagonist, W7, inhibited dissociation of K-RasB. Addition of platelet or MCF-7 cytosol alone to isolated platelet membranes did not cause dissociation of K-RasB and only addition of exogenous CaM caused dissociation. The results suggest a potential role for Ca2+/CaM in the regulation of K-RasB function. PMID- 12727205 TI - Compromised calnexin function in calreticulin-deficient cells. AB - Calnexin and calreticulin are molecular chaperones, which are involved in the protein folding, assembly, and retention/retrieval. We know that calreticulin deficiency is lethal in utero, but do not understand the contribution of chaperone function to this phenotype. Here we studied protein folding and chaperone function of calnexin in the absence of calreticulin. We show that protein folding is accelerated and quality control is compromised in calreticulin deficient cells. Calnexin-substrate association is severely reduced, leading to accumulation of unfolded proteins and a triggering of the unfolded protein response (UPR). PERK and Ire1alpha and eIF2alpha are also activated in calreticulin-deficient cells. We show that the absence of calreticulin can have devastating effects on the function of the others, compromising overall quality control of the secretory pathway and activating UPR-dependent pathways. PMID- 12727206 TI - Cloning of zebrafish BAD, a BH3-only proapoptotic protein, whose overexpression leads to apoptosis in COS-1 cells and zebrafish embryos. AB - The BH3-only proapoptotic protein, BAD, was cloned from zebrafish embryos and its properties were characterized. Zebrafish BAD (zBAD) is a protein with 147 amino acids that contains a BH3 domain and a putative 14-3-3 binding site with the sequence of RPRSRS(84)AP, corresponding to S(136) in mouse BAD (mBAD). zBAD shares 34%, 28%, and 29% amino acid sequence identity to the human, mouse, and rat BAD, respectively. RT-PCR analysis revealed that the expression of zBAD gene is found in various parts of zebrafish tissues. The treatment with the z-VAD fmk, a broad-range caspase inhibitor, in COS-1 cells significantly increased the expression of zebrafish BAD fusion proteins (GFP-zBAD and HA-zBAD), indicating that zebrafish BAD fusion proteins may be cleaved by caspase(s). zBAD was shown to induce apoptosis when it was overexpressed in COS-1 cells. In addition, zBAD was also expressed in muscle cells under the muscle-specific promoter from zebrafish alpha-actin gene. Abnormality in the skeletal muscles and the loss of green fluorescence signal in the same region were observed. Taken together, our results indicate that zBAD could induce apoptosis in vitro and in vivo and may have biological implications in apoptosis during zebrafish development. PMID- 12727208 TI - Altering the substrate chain-length specificity of an alpha-glucosidase. AB - Dextran glucosidases show high sequence identity (50%) to Bacillus sp. SAM1606 alpha-glucosidase, which is more specific for short-chain substrates. Sequence comparison of these enzymes as well as molecular modeling studies predicted that the extension of loop 4 of the (beta/alpha)(8)-barrel fold may be responsible for the narrower specificity of SAM1606 alpha-glucosidase with respect to substrate chain length. Indeed, deletion mutants of SAM1606 alpha-glucosidase that lack this extension showed higher relative activities toward dextran and long-chain isomaltooligosaccharides. Kinetic and thermodynamic analyses of oligosaccharide hydrolysis catalyzed by SAM1606 alpha-glucosidase and its deletion mutants suggested that the loss of such extension(s) in loop 4 should energetically destabilize the Michaelis complexes with long-chain substrates to result in smaller differences between the activation free energies for the enzymatic hydrolyses of isomaltoheptaose and isomaltose than those observed for the wild type enzyme. This is the reason that dextran glucosidase, whose loop 4 is shorter in length, shows broader substrate chain-length specificity than does SAM1606 alpha-glucosidase. PMID- 12727207 TI - Nuclear translocation of Xenopus laevis paxillin. AB - Paxillin has been recognized as a focal adhesion adapter protein that participates in the integrin-mediated signaling. An earlier study [Ogawa et al. Biochim. Biophys. Acta 1519 (2001) 235] found that frog paxillin was expressed in the kidney epithelial cell line A6 and localized in the nucleus. Here, in this study, we have found that the expression of frog paxillin is up-regulated in the S phase of cell cycle. The protein became phosphorylated on tyrosine when the cells were grown on vitronectin; the tyrosine phosphorylation was not detectable when the cells were cultured on fibronectin, laminin or poly-D-lysine. On the other hand, MAP kinase was revealed to phosphorylate frog paxillin on serine. Both phosphorylation events, namely on tyrosine and serine, were essential for the nuclear translocation of this protein. Our results suggest that the integrin mediated signaling pathway and the MAP kinase pathway meet at paxillin. PMID- 12727209 TI - Differentiation of bone marrow cells into cells that express liver-specific genes in vitro: implication of the Notch signals in differentiation. AB - Bone marrow (BM) stem cells have been shown to differentiate into liver cells. It remains difficult to sort and culture BM stem cells, and the gene expression of liver-specific proteins in these cells has not been fully investigated. We used a negative selective magnetic cell separation system to obtain stem cell-enriched BM cells. The cells obtained were cultured with hepatocytes or with hepatocyte growth factor (HGF), and the differentiation of BM cells into cells expressing liver-specific genes, hepatocyte nuclear factor (HNF) 1alpha, cytokeratin (CK) 8, alpha-fetoprotein (AFP), and albumin was investigated by the reverse transcription-polymerase chain reaction. We also investigated the gene expressions of Notch receptor-1 (Notch-1) and its ligand Jagged-1 in BM cell differentiation. Sorted BM cells showed positive for Sca-1 (Ataxin-1) by immunofluorescence staining. Fluorescence activated cell sorter analysis showed that 32.6% of sorted BM cells had a high level of expression of the hematopoietic stem cell marker CD90 (Thy-1). When cultured with hepatocytes, these cells expressed the liver-specific genes HNF1alpha and CK8 on culture day 3, AFP and albumin on culture day 7. When cultured with HGF (20ng/ml), the cells expressed HNF1alpha on day 3 and CK8 on day 7. Gene expressions of Notch-1 and Jagged-1 were detected in cultured BM cells on day 3. These results suggest that the negative selective magnetic cell separation system is useful for the rapid preparation of stem cell-enriched BM cells, and that the Notch signaling pathway plays a role in BM cell differentiation into a hepatocyte lineage in vitro. PMID- 12727210 TI - TPA activates p21WAF-1 promoter in human T-cells through its second most upstream Sp1 site. AB - The p21(WAF-1) promoter contains binding sites for a number of transcription factors which mediate its activation by a variety of external signals. Moreover, it has been reported that the transcription factors involved in p21(WAF-1) activation by certain signaling factors, like the phorbol ester TPA, may vary in different cell types. We were interested in elucidating the mechanism of p21(WAF 1) activation by TPA in human T-cells, since this activation could explain the antagonistic effect of PKC on apoptosis induction in these cells noted in our previous studies. Using the Jurkat human T-cells we found that TPA activated p21(WAF-1) expression by a PKC-dependent mechanism and that out of six Sp1 binding sites residing in its promoter the second most upstream one was critically essential for this activation. Since p21(WAF-1) is known to inhibit the onset of apoptosis, its PKC-dependent activation may likely account for the PKC antagonistic effect on apoptosis induction in these cells. PMID- 12727211 TI - Human follicular dendritic cells and fibroblasts share the 3C8 antigen. AB - Although the pivotal role of follicular dendritic cells (FDCs) in humoral immune responses has been demonstrated, little is known of the molecular basis of biological functions and the cellular origin of FDC. We have recently generated a monoclonal antibody (mAb) against FDC by immunizing mice with FDC-like tonsillar stromal cells. The mAb 3C8 does not cross-react with bone marrow-derived blood cells. Partial amino acid sequencing revealed that 3C8 Ag is a novel human protein. In this study, we carried out a detailed analysis of 3C8 immunoreactivity with tonsil sections to examine cellular distribution of 3C8 Ag. 3C8 Ab recognized connective tissue fibroblasts in addition to FDC. Western blot analysis indicated that 3C8 antigen is expressed in various fibroblasts and is specific to human species. Furthermore, there was a correlation between 3C8 expression in several stromal cell lines and their co-stimulatory activity of germinal center B cell proliferation. These findings strongly support the view that FDCs originate from local fibroblasts. PMID- 12727212 TI - In vivo processed fragments of IGF binding protein-2 copurified with bioactive IGF-II. AB - Proteolysis of insulin-like growth factor binding proteins (IGFBPs), the major carrier of insulin-like growth factors (IGFs) in the circulation, is an essential mechanism to regulate the bioavailability and half-live of IGFs. Screening for peptides in human hemofiltrate, stimulating the survival of PC-12 cells, resulted in the isolation of C-terminal IGFBP-2 fragments and intact IGF-II co-eluting during the chromatographic purification procedure. The IGFBP-2 fragments exhibited molecular masses of 12.7 and 12.9kDa and started with Gly169 and Gly167, respectively. The fragments were able to bind both IGFs. The stimulatory effect of the purified fraction on the survival of the PC-12 cells could be assigned exclusively to IGF-II, since it was abolished by the addition of neutralizing IGF-II antibodies. We suggest that in the circulation IGF-II is not only complexed with intact IGFBP but also with processed IGFBP-2 fragments not impairing the biological activity of IGF-II. PMID- 12727213 TI - PKC is required for activation of ROCK by RhoA in human endothelial cells. AB - Rho/Rho-kinase (ROCK) complex formation is the only proposed mechanism for ROCK activation. Rho/ROCK and PKC can exhibit a convergence of cellular effects such as suppression of endothelial nitric oxide synthase (eNOS) expression. We, therefore, investigated the role of PKC in RhoA/ROCK complex formation and activation linked to eNOS expression in cultured human umbilical vein endothelial cells. We showed that expression of constitutively active RhoA (Rho63) or ROCK (CAT) suppressed eNOS gene expression. This effect of Rho63 but not that of CAT was abolished by phorbol ester-sensitive PKC depletion. Accordingly, depletion or inhibition of PKC prevented ROCK activation by Rho63 without affecting RhoA/ROCK complex formation. Similarly, suppression of eNOS expression and activation of ROCK, but not RhoA by thrombin were prevented by PKC inhibition or depletion. These results indicate that RhoA/ROCK complex formation alone is not sufficient and PKC is required for RhoA-induced ROCK activation leading to eNOS gene suppression. PMID- 12727214 TI - Chronic glucocorticoid receptor activation impairs CREB transcriptional activity in clonal neurons. AB - Excessive circulating levels of glucocorticoids are thought to be associated with cognitive impairment. We provide evidence that chronic activation of the glucocorticoid receptor (GR) in clonal neurons inhibits the transcriptional activity of the cyclic AMP response element-binding protein (CREB), which is believed to be involved in memory processes. To investigate the underlying mechanism we studied the phosphorylation of CREB and found altered phosphorylation kinetics in neurons chronically treated with glucocorticoids. Our results demonstrate a hitherto unrecognized crosstalk between the cyclic AMP and glucocorticoid pathway and may provide the molecular basis for the effects of long-term glucocorticoid exposure on cognitive function. PMID- 12727215 TI - Kallikrein-like prorenin-converting enzymes in inbred hypertensive mice. AB - Kallikreins are a group of serine proteases and are distinguished by having serine residue at their active site. Their general function is to convert inactive pro-peptide into its biologically active form. In recent years, emerging evidence indicates that some kallikrein-kinin enzymes also play a role in the modulation of renin-angiotensin system. These kallikrein-like prorenin converting enzymes act on renin-angiotensin by converting prorenin into biologically active renin. In this investigation, kallikrein-like prorenin converting enzyme (PRCE C) (mK9) is isolated from genetically inbred high blood pressure (BPH) and their normal counterparts (BPN) mice, and its protein levels are quantitated. Levels of mRNA expression are also compared. Additionally, localization of the enzyme is visualized by in situ hybridization histochemistry. Results indicated higher levels of PRCE C (mK9) enzyme in BPH mice in comparison to their normal counterparts. mRNA expression was also higher in BPH mice. In situ hybridization histochemistry results localized PRCE C (mK9) in the striated duct cells of submandibular gland. PMID- 12727216 TI - p44/42 MAPK activation is necessary for receptor activator of nuclear factor kappaB ligand induction by high extracellular calcium. AB - Although extracellular calcium (Ca(2+)(o)) has been suggested to modulate bone remodeling, the exact mechanism is unclear. This study was performed to explore the signaling pathways of high Ca(2+)(o) that are responsible for controlling the expression of receptor activator of NF-kappaB ligand (RANKL) in mouse osteoblastic cells. As previously reported, high Ca(2+)(o) increased RANKL expression. However, the G protein-coupled Ca(2+)(o)-sensing receptor (CaSR) was not detected in the primary cultured mouse osteoblastic cell. The inhibition of the pertussis-sensitive G protein, phospholipase C, protein kinase C, intracellular calcium mobilization, p38 MAPK, or phosphoinositide 3-kinase did not block RANKL induction caused by high Ca(2+)(o). In contrast, the inhibition of p44/42 MAPK pathway reduced the RANKL expression induced by high Ca(2+)(o). Moreover, high Ca(2+)(o) activated p44/42 MAPK and MEK1/2. These results suggest that RANKL induction by high Ca(2+)(o) might not be mediated by CaSR and its putative downstream signaling pathways, but the pathway employing p44/42 MAPK is involved in the high Ca(2+)(o)-induced RANKL expression in mouse osteoblastic cells. PMID- 12727217 TI - Mechanism of pH regulation of connexin 43 expression in MC3T3-E1 cells. AB - Gap junction (GJ) expression and function allowing cell-cell communication among osteoblasts may be important in new bone formation. An alkaline milieu stimulates mineralization, while extracellular acidification leads to demineralization of bone. It was previously demonstrated that alkaline pH increases, while acid pH decreases GJ intercellular communication by an increase in steady-state GJ connexin 43 (Cx43) mRNA and protein expression. At pH 7.6, transcription of new Cx43 mRNA was significantly higher than that at pH 6.9 but not significantly different at pH 7.2, as assessed by nuclear run-on assay. Transcription of new Cx43 mRNA was higher at pH 7.2 compared to that at pH 6.9. Although Cx43 mRNA half-life tended to be longer at pH 7.6, analysis of variance did not yield a significant difference of the Cx43 mRNA half-life at any of the pHs tested. Likewise, the half-life of Cx43 protein at pHs of 6.9, 7.2, and 7.6 was not significantly different. Plasma membrane and cytosolic Cx43 fractions were proportionately similar at pH 7.2 and 6.9. Thus, the decrease in Cx43 mRNA at low pH compared to high pH is due to a decrease in the transcription rate of Cx43 but not due to an alteration of message stability. The early uncoupling of gap junctions by low pH found previously does not appear to be due to changes in the half-life nor distribution of Cx43 protein between cytosolic and plasma membrane compartments. PMID- 12727218 TI - Inhibition of endothelial cell proliferation by the recombinant kringle domain of tissue-type plasminogen activator. AB - Tissue-type plasminogen activator (tPA) is a multidomain serine protease that converts the zymogen plasminogen to plasmin. tPA contains two kringle domains which display considerable sequence identity with those of angiostatin, an angiogenesis inhibitor. TK1-2, a recombinant kringle domain composed of t-PA kringles 1 and 2 (Ala(90)-Thr(263)), was produced by both bacterial and yeast expression systems. In vitro, TK1-2 inhibited endothelial cell proliferation stimulated by basic fibroblast growth factor, vascular endothelial growth factor, and epidermal growth factor. It did not inhibit proliferation of non-endothelial cells. TK1-2 also inhibited in vivo angiogenesis in the chick embryo chorioallantoic membrane model. These results suggest that the recombinant kringle domain of t-PA is a selective inhibitor of endothelial cell growth and identifies this molecule as a novel anti-angiogenic agent. PMID- 12727220 TI - Copper-induced oxidation of epinephrine: protective effect of D-DAHK, a synthetic analogue of the high affinity copper binding site of human albumin. AB - Epinephrine is known to be rapidly oxidized during sepsis. Ischemia and acidosis, which often accompany sepsis, are associated with the release of weakly bound cupric ions from plasma proteins. We investigated whether copper promotes oxidation of epinephrine at both physiological and acidic pH and whether D-Asp-D Ala-D-His-D-Lys (D-DAHK), a human albumin (HSA) N-terminus synthetic peptide with a high affinity for cupric ions, attenuates this oxidation. Epinephrine alone [100 microM] or with CuCl(2) [10 microM], and with CuCl(2) [10 microM] and D-DAHK [20 microM] at pH 7.4, 7.0, 6.5, and 6.0 were incubated for 1h at 37 degrees C. Epinephrine oxidation was measured by the spectrophotometric quantification of its oxidation product, adrenochrome. We found that adrenochrome increased, suggesting copper-induced oxidation of epinephrine. At pH 7.4, 7.0, 6.5, and 6.0, adrenochrome increased by 47%, 53%, 24%, and 6% above baseline, respectively. D DAHK attenuated the copper-induced oxidation of epinephrine to baseline levels. These in vitro results indicate that copper-induced epinephrine oxidation is greatest at the physiological pH 7.4 as well as in severe acidosis, pH 7.0, and that D-DAHK completely inhibits this oxidation. PMID- 12727219 TI - Structure, tissue expression pattern, and function of the amino acid transporter rat PAT2. AB - The second member of the PAT (proton-coupled amino acid transporter) family of H(+)-coupled, pH-dependent, Na(+)-independent amino acid transporters was isolated from a rat lung cDNA library. The cDNA for rat PAT2 is 2396bp in length, including 70bp of 5'UTR and a poly(A) tail. The transporter gene, consisting of 10 exons, is located on rat chromosome 10q22. The cDNA codes for a protein of 481 amino acids with 72% identity (over 449 amino acids) with rat PAT1. Tissue expression studies demonstrate that mRNA abundance is generally low with highest levels being detected in lung and spleen, with lower levels in the brain, heart, kidney, and skeletal muscle. Functional expression in either mammalian cells or Xenopus laevis oocytes demonstrates that rat PAT2 mediates pH-dependent, Na(+) independent uptake of glycine, proline, and alpha(methyl)aminoisobutyric acid (MeAIB). In conclusion PAT2 has a limited tissue distribution, higher affinity (Michaelis-Menten constant for glycine uptake between 0.49 and 0.69mM), and distinct substrate specificity compared to PAT1. PMID- 12727221 TI - Cyclic AMP-dependent protein kinase A and CREB are involved in neuregulin-induced synapse-specific expression of acetylcholine receptor gene. AB - Neuregulin is reported to stimulate synapse-specific transcription of acetylcholine receptor (AChR) genes in the skeletal muscle fiber by multiple signaling pathways such as ERK, PI3K, and JNK. The co-localization of PKA mRNA with AChR and ErbBs, receptors for neuregulin, at the confined region of synapse implicates the putative role of PKA in neuregulin-induced AChR gene expression. In the present study, we found that mRNA and protein of a regulatory subunit of PKA (PKARIalpha) were concentrated at synaptic sites of the rat sternomastoid muscle fiber, while those of ERK and PI3K were uniformly distributed throughout the muscle fiber. Neuregulin (100 ng/ml) increased both PKA activity in the nucleus and AChRdelta subunit gene transcription in cultured Sol8 myotubes. These increases were significantly blocked by a specific PKA inhibitor H-89 (100 nM) and an adenylcyclase inhibitor SQ 22536 (200 microM) (72.5% and 60.1%, respectively). Furthermore, neuregulin phosphorylated CREB, a well-known down stream transcription factor of PKA. While H-89 inhibited CREB phosphorylation, H 89 and PD098059 (50 microM), a specific MEK1/2 inhibitor, did not inhibit the phosphorylation of ERK and CREB, respectively, suggesting no cross-talk between PKA and ERK pathways. In conclusion, neuregulin increases AChRdelta subunit gene transcription, in part, by the activation of PKA/CREB, an alternative route to the previously reported ERK signaling pathway. PMID- 12727223 TI - A high-throughput screen for single gene activities: isolation of apoptosis inducers. AB - We describe a novel genetic screen that is performed by transfecting every individual clone of an expression library into a separate population of cells in a high-throughput mode. The screen allows one to achieve a hitherto unattained sensitivity in expression cloning which was exploited in a first read-out to clone apoptosis-inducing genes. This led to the isolation of several genes whose proteins induce distinct phenotypes of apoptosis in 293T cells. One of the isolated genes is the tumor suppressor cytochrome b(L) (cybL), a component of the respiratory chain complex II, that diminishes the activity of this complex for apoptosis induction. This gene is more efficient and specific for causing cell death than a drug with the same activity. These results suggest further applications, both of the isolated genes and the screen. PMID- 12727222 TI - Overexpression of oxidized protein hydrolase protects COS-7 cells from oxidative stress-induced inhibition of cell growth and survival. AB - Oxidized protein hydrolase (OPH) preferentially degrades oxidatively damaged proteins in vitro and is widely distributed in various cells and tissues. The role of OPH in intact cells exposed to oxidative stress was examined. For this purpose, using COS-7, a cell line derived from African green monkey kidney, COS-7 OPH cells that stably overexpressed OPH were established. When COS-7-OPH cells were exposed to oxidative stress induced by H(2)O(2) and paraquat, accumulation of protein carbonyls in the cells was apparently lower than that of parental COS 7 cells, and COS-7-OPH cells were significantly resistant to the oxidative stress compared with parental COS-7 cells. The majority of overexpressed OPH in the cells was found to be located uniformly in cytosol, and its location was not altered by H(2)O(2)-induced oxidative stress. Above results indicate that OPH in intact cells plays a preventive role against oxidative stress and suggest that OPH relieves cells from accumulation of oxidatively damaged proteins. PMID- 12727224 TI - Cloning of rat ABCA7 and its preferential expression in platelets. AB - We cloned the full-length cDNA of a rat orthologue of ABCA7 (rABCA7) from rat platelets. The cDNA of rABCA7 is 6510bp in length and encodes a protein of 2170 amino acids. The amino acid sequence of rABCA7 exhibits homology to those of mouse ABCA7 (92.5% identical in amino acid sequence) and human ABCA7 (76.6%). We obtained two clones of monoclonal antibodies against rABCA7 recognizing different epitopes. Analysis of CHO cells stably expressing rABCA7 by confocal laser scanning microscopy indicated that rABCA7 is mainly located in the plasma membrane. Western blot analysis of rat tissues revealed that rABCA7 was preferentially expressed in platelets and that its apparent molecular mass was 250kDa. This is the first report of the tissue distribution of rABCA7 at the protein level and is the first reported case of ABC transporters being expressed in platelets, suggesting their important role in platelet function. PMID- 12727225 TI - Synthetic propeptide inhibits mosquito midgut chitinase and blocks sporogonic development of malaria parasite. AB - Incessant transmission of the parasite by mosquitoes makes most attempts to control malaria fail. Blocking of parasite transmission by mosquitoes therefore is a rational strategy to combat the disease. Upon ingestion of blood meal mosquitoes secrete chitinase into the midgut. This mosquito chitinase is a zymogen which is activated by the removal of a propeptide from the N-terminal. Since the midgut peritrophic matrix acts as a physical barrier, the activated chitinase is likely to contribute to the further development of the malaria parasite in the mosquito. Earlier it has been shown that inhibiting chitinase activity in the mosquito midgut blocked sporogonic development of the malaria parasite. Since synthetic propeptides of several zymogens have been found to be potent inhibitors of their respective enzymes, we tested propeptide of mosquito midgut chitinase as an inhibitor and found that the propeptide almost completely inhibited the recombinant or purified native Anopheles gambiae chitinase. We also examined the effect of the inhibitory peptide on malaria parasite development. The result showed that the synthetic propeptide blocked the development of human malaria parasite Plasmodium falciparum in the African malaria vector An. gambiae and avian malaria parasite Plasmodium gallinaceum in Aedes aegypti mosquitoes. This study implies that the expression of inhibitory mosquito midgut chitinase propeptide in response to blood meal may alter the mosquito's vectorial capacity. This may lead to developing novel strategies for controlling the spread of malaria. PMID- 12727226 TI - Highly potent propargylamine and allylamine inhibitors of bovine plasma amine oxidase. AB - Propargylamine was reported many years ago to be a mechanism-based inhibitor of bovine plasma amine oxidase (BPAO), though the potency was modest and allylamine was a substrate. Herein, selected 3-substituted propargylamines and allylamines were found to be potent time-dependent inactivators of BPAO, exhibiting IC(50) values of 2-13 microM at 30 degrees C, making them the most potent BPAO inhibitors reported to date. The most potent compound, trans-3-chloroallylamine, was previously found not to inhibit the flavin-dependent monoamine oxidase (the cis isomer did), and thus appears to be a highly selective inhibitor. PMID- 12727228 TI - Specific blockade of the ERK pathway inhibits the invasiveness of tumor cells: down-regulation of matrix metalloproteinase-3/-9/-14 and CD44. AB - Elevated expression of matrix metalloproteinases (MMPs) is associated with increased metastatic potential in many tumor cells. As activation of the ERK pathway has been linked to the expression of MMP-9, we examined a possible correlation between ERK activation, MMP-9 expression, and invasive phenotype in human tumor cells. Activation state of the ERK pathway in tumor cells was well correlated with the invasive phenotype, which was determined by the ability of cells to invade through reconstituted extracellular matrix. Elevated expression of MMP-9 as well as of MMP-3, MMP-14, and CD44 was observed in tumor cells in which constitutive activation of the ERK pathway is detected. Blockade of the ERK pathway by treatment with PD184352, a specific and powerful inhibitor of mitogen activated protein (MAP) kinase/ERK kinase (MEK), suppressed the expression of MMP 3, MMP-9, MMP-14, and CD44, and inhibited markedly the invasiveness of tumor cells. These results imply that, in addition to anti-proliferative effects, specific blockade of the ERK pathway is expected to result in anti-metastatic effects in tumor cells. PMID- 12727227 TI - Induction of neurite outgrowth in PC12 cells by the bacterial nucleoside N6 methyldeoxyadenosine is mediated through adenosine A2a receptors and via cAMP and MAPK signaling pathways. AB - We have previously shown that N(6)-methyldeoxyadenosine (MDA) is an inducer of differentiation in several tumor cells. Here we show that in addition to its ability to induce neurite-outgrowth in PC12 cells, MDA also significantly enhances the nerve-growth factor-mediated neurite outgrowth of these cells. Thus, MDA acts synergistically with NGF to repress cdc2 and cdk2 synthesis and to enhance tyrosine hydroxylase synthesis. To further elucidate the mechanisms of action of MDA, we investigated the effect of this drug on various signaling pathways. The neuritogenesis observed in PC12 following MDA treatment is mediated through activation of adenylyl cyclase in a PKA independent process and through the recruitment of the p44/p42 MAPK pathway. Furthermore, the adenosine A(2a) receptor antagonist ZM 241385 prevents the MDA-induced neuritogenesis, suggesting that MDA mediates its effect via this adenylyl cyclase-coupled A(2a) receptor. Collectively, these findings suggest that, in PC12 cells, the MDA-induced neuritogenesis requires the recruitment of adenosine A(2a) receptor, the stimulation of adenylate cyclase, and the activation of the p44/42MAP kinase cascade. PMID- 12727229 TI - Effects of iron and oxygen species scavengers on Listeria spp. chemiluminescence. AB - Listeria monocytogenes and Listeria innocua are able, under certain conditions, to produce chemiluminescence (CL), which is amplified by luminol. Kinetic studies of CL by L. monocytogenes and L. innocua show a close parallelism between CL and growth curves during the exponential phase, with a maximum of CL reached just before entrance of bacteria into the stationary phase. CL is tightly correlated with the release of oxygen compounds. The reactive oxygen species scavengers tryptophan, mannitol, and tiron, as well as cellobiose and high temperature, were assessed with regard to CL in the two Listeria species. Only tiron strongly reduced the CL emitted by L. monocytogenes and L. innocua. On the other hand, charcoal pretreatment of the growth medium inhibited the CL, whereas ferric citrate strongly increased the CL of L. monocytogenes and L. innocua. These data suggest that iron and superoxide radical are implicated in the CL produced by these bacteria, but this phenomenon is not correlated to virulence. PMID- 12727230 TI - Diet-dependent obesity and hypercholesterolemia in the New Zealand obese mouse: identification of a quantitative trait locus for elevated serum cholesterol on the distal mouse chromosome 5. AB - AIMS: New Zealand obese (NZO) mice exhibit a polygenic syndrome of obesity, insulin resistance, and hypercholesterolemia that resembles the human metabolic syndrome. This study was performed in order to locate genes responsible for elevated serum cholesterol and to compare their effects under a standard and high fat diet. METHODS: A backcross population of NZO with SJL mice (NZO x F1(SJL x NZO)) was generated. Mice were raised on a normal or high fat diet and were monitored for 22 weeks (body weight, serum cholesterol, and blood glucose). A genome-wide scan was performed by genotyping of approximately 200 polymorphic microsatellite markers by PCR and linkage analysis was performed with the MAPMAKER program. RESULTS: In the genome-wide scan, a single susceptibility locus for hypercholesterolemia (Chol1/NZO, maximum LOD score 14.5 in a combined population of 523 backcross mice) was identified on chromosome 5. Cholesterol levels were significantly elevated in both male and female homozygous carriers of the Chol1/NZO allele. The locus maps 40cM distal of the previously described obesity locus Nob1 in the vicinity of the marker D5Mit244 and in the vicinity of hypercholesterolemia QTL previously identified in the NZB, CAST, and C57BL/6J strains. Chol1/NZO was not associated with elevated body weight, serum insulin, or hyperglycemia. The high fat diet significantly increased serum cholesterol levels, but the fat content of the diet did not alter the absolute effect of Chol1/NZO. CONCLUSIONS: Chol1/NZO is a major susceptibility locus on the distal mouse chromosome 5, which produces gender-independent hypercholesterolemia in NZO mice. The effect of Chol1/NZO was independent of the dietary fat content and was not associated with the other traits of the metabolic syndrome. Thus, it is suggested that the responsible gene might be involved in cholesterol metabolism. PMID- 12727231 TI - Lysozyme conjugate immune complex formation and the effects on substrate hydrolysis. AB - The defined estrone glucuronide-lysozyme conjugate E3, that is acylated solely at K33, was used as a probe for the steric requirements of the active site cleft of chicken type lysozymes. When the immune complex was formed with an anti-estrone glucuronide antiserum, the rate of lysis of the E3 conjugate with the large bacterial substrate Micrococcus lysodeikticus was inhibited by over 90%. However, when the small hexamer of N-acetyl glucosamine was used as the substrate, the rate of hydrolysis by the immune complex was accelerated by 350% compared with the control rate. Thus, inhibition by the anti-estrone glucuronide cannot be caused simply by steric occlusion of the active site. Other factor(s) in the immune complex activate the hydrolysis reaction, most likely by favouring the conformations that lead to the transition state. PMID- 12727232 TI - Identification of a copper chaperone from tomato fruits infected with Botrytis cinerea by differential display. AB - Differential display was used to isolate tomato genes responding to fungal infection. Here we describe the isolation and characterization of a gene that is down-regulated in tomato fruits infected with the phytopathogen Botrytis cinerea. The cDNA identified encodes a protein that shares sequence similarity to the amino terminal region of CCH, a copper chaperone from Arabidopsis thaliana, that participates in intracellular copper homeostasis by delivering Cu to the secretory pathway. The fact that this newly characterized tomato gene, referred to as LeCCH (Lycopersicon esculentum copper chaperone), be differentially expressed after fungal infection, suggests an interesting relationship between copper homeostasis and plant defense responses. LeCCH contains the conserved metal-binding domain MXCXGC but interestingly, lacks the C-terminal extension present in previously described plant members of this copper chaperone family, that seems to be involved in metallochaperone intercellular transport. This fact indicates that LeCCH is a novel plant copper chaperone that could act locally at the infection site, affecting the copper homeostasis in this particular stress situation. PMID- 12727233 TI - Proteomics in Drosophila melanogaster: first 2D database of larval hemolymph proteins. AB - A proteomic approach was used for the identification of larval hemolymph proteins of Drosophila melanogaster. We report the initial establishment of a two dimensional gel electrophoresis reference map for hemolymph proteins of third instar larvae of D. melanogaster. We used immobilized pH gradients of pH 4-7 (linear) and a 12-14% linear gradient polyacrylamide gel. The protein spots were silver-stained and analyzed by nanoLC-Q-Tof MS/MS (on-line nanoscale liquid chromatography quadrupole time of flight tandem mass spectrometry) or by Matrix assisted laser desorption time of flight MS (MALDI-TOF MS). Querying the SWISSPROT database with the mass spectrometric data yielded the identity of the proteins in the spots. The presented proteome map lists those protein spots identified to date. This map will be updated continuously and will serve as a reference database for investigators, studying changes at the protein level in different physiological conditions. PMID- 12727235 TI - Effect of NaCl on thermophilic (55 degrees C) methanol degradation in sulfate reducing granular sludge reactors. AB - The effect of NaCl on thermophilic (55 degrees C) methanol conversion in the presence of excess of sulfate (COD/SO(4)(2-)=0.5) was investigated in two 6.5L lab-scale upflow anaerobic sludge bed reactors inoculated with granular sludge previously not adapted to NaCl. Methanol was almost completely used for sulfate reduction in the absence of NaCl when operating at an organic loading rate of 5 g CODL(-1)day(-1) and a hydraulic retention time of 10h. The almost fully sulfidogenic sludge consisted of both granules and flocs developed after approximately 100 days in both reactors. Sulfate reducing bacteria (SRB) outcompeted methane producing archaea (MPA) for methanol, but acetate represented a side-product, accounting for maximal 25% of the total COD converted. Either MPA or SRB did not use acetate as substrate in activity tests. High NaCl concentrations (25 gL(-1)) completely inhibited methanol degradation, whereas low salt concentrations (2.5 g NaClL(-1)) provoked considerable changes in the metabolic fate of methanol. The MPA were most sensitive towards the NaCl shock (25 gL(-1)). In contrast, the addition of 2.5 gL(-1) of NaCl stimulated MPA and homoacetogenic bacteria. PMID- 12727234 TI - Microbial diversity in a thermophilic aerobic biofilm process: analysis by length heterogeneity PCR (LH-PCR). AB - A two-stage pilot-scale thermophilic aerobic suspended carrier biofilm process (SCBP) was set up for the on-site treatment of pulp and paper mill whitewater lining. The microbial diversity in this process was analyzed by length heterogeneity analysis of PCR-amplified 16S ribosomal DNA. The primer pair selected for PCR amplification was first evaluated by a computational analysis of fragment lengths in ten main phylogenetical eubacterial groups. The fragment contained the first third of the 16S rRNA gene, which was shown to vary naturally between 465 and 563 bp in length. The length heterogeneity analysis of polymerase chain reaction (LH-PCR) profile of the biomass attached to carrier elements was found to be diverse in both stages of the SCBP. During normal operating conditions, sequences belonging to beta-Proteobacteria, Cytophaga/Flexibacter/Bacteroides group and gamma-Proteobacteria were assigned to the most prominent LH-PCR peak. Samples from the suspended biomass consisted of completely different bacterial populations, which were, however, similar in the serial reactors. The pilot process experienced alkaline shocks, after which Bacillus-like sequences were detected in both the biofilm and suspended biomass. However, when the conditions were reversed, the normal microbial population in the biofilm recovered rapidly without further biomass inoculations. This study shows that LH-PCR is a valuable method for profiling microbial diversity and dynamics in industrial wastewater processes. PMID- 12727236 TI - Nitrification efficiency and nitrifying bacteria abundance in combined AS-RBC and A2O systems. AB - This study makes a comparison between the nitrification performance of TNCU-I (a combined activated sludge-rotating biological contactor process) and A2O systems by the use of a pilot plant and batch experiments. The nitrifier abundance in both systems was determined, using cloning-denaturing gradient gel electrophoresis (DGGE) and fluorescent in-situ hybridization (FISH), to investigate the role of rotating biological contactor in the TNCU-I process. The stability of the nitrification performance and the specific nitrification rate were found to be greater in TNCU-I system than in the A2O system. RBC biofilm promoted nitrifying activity that contributed to the nitrification performance, especially at a low SRT. By using the cloning-DGGE method, the genera Nitrosospira and Nitrospira were found to be present in all the samples, while the genus Nitrosomonas was observed only in the TNCU-I RBC biofilm. In addition, the proportions of ammonia oxidizer in the TNCU-I RBC biofilm, the TNCU-I activated sludge and the A2O activated sludge were 11.4%, 13.2%, and 4.1%, respectively, higher than the nitrite oxidizer fractions of 3.3%, 5.7% and 2.1%, respectively, according to the cloning-DGGE method. On the other hand, the proportions of ammonia oxidizers in the afore-mention materials were 10.3%, 13.7%, and 5.2%, higher than the nitrite oxidizer fractions of 2.5%, 3.6% and 2.3%, according to the FISH experiments. This implies that the proportion of ammonia oxidizer in the TNCU-I process was 3.2 and 2.6 times that in the A2O process, determined by the cloning-DGGE and FISH methods, respectively. These amounts are also close to the ammonia oxidization rate of 2.9 times. All the data show that RBC added to the aerobic zone of TNCU-I process would increase the nitrifier abundance and enhance the nitrification performance of the system. PMID- 12727237 TI - Comparative electrochemical inactivation of bacteria and bacteriophage. AB - Electric fields and currents have been shown to be capable of disinfecting drinking water and reducing the numbers of bacteria and yeast in food. However, little research has been conducted regarding the effectiveness of electric fields and currents in the inactivation of viruses. The objective of this study was to compare the ability of bacteria and bacteriophage to survive exposure to direct electric current in an electrochemical cell, where they would be subject to irreversible membrane permeabilization processes, direct oxidation of cellular/viral constituents by electric current, and disinfection by electrochemically generated oxidants. Suspensions of the bacteria Escherichia coli and Pseudomonas aeruginosa and bacteriophage MS2 and PRD1 at both high (approximately 1 x 10(6)CFU or PFU/mL) and low (approximately 1 x 10(3)CFU or PFU/mL) population densities were exposed to currents ranging from 25 to 350 mA in 5s pulses. Post-exposure plaque counts of the bacteriophage were proportionally higher than bacterial culturable counts at corresponding current exposures. E. coli and MS2 were then exposed to 5 mA for 20 min at both high and low population densities. The inactivation rate of E. coli was 2.1-4.3 times greater than that of MS2. Both bacteria and bacteriophage were more resistant to exposure to direct current at higher population densities. Also, amelioration of inactivation within the electrochemical cell by the reducing agent glutathione indicates the major mechanism of inactivation in the electrochemical cell is disinfection by electrochemically generated oxidants. The implications of these results are that technologies relying upon direct current to reduce the numbers of microbes in food and water may not be sufficient to reduce the numbers of potentially pathogenic viruses and ensure the safety of the treated food or water. PMID- 12727238 TI - Biosorption of humic and fulvic acids to live activated sludge biomass. AB - Biosorption of high molecular weight humic substances (HS) to activated sludge (AS) biomass may be considered as a preliminary step previous to enzymatic hydrolysis breakdown and biological uptake. Two standard HS, Suwannee River humic and fulvic acids, were biosorbed onto live AS biomass collected from full-scale wastewater treatment plants. Biosorption isotherms were corrected for interference from organic matter desorbed from AS biomass. The effect of pH, calcium and ionic strength on biosorption was tested. HS biosorption to live AS biomass obeyed the Freundlich isotherm equation. Biosorption increased with decreasing pH, increasing calcium and ionic strength concentration. Higher biosorption at low pH may be attributed to hydrophobic interactions between HS and AS biomass extracellular polymers (EPS). Hydrophobic and cationic bridging effects between HS and AS EPS were the mechanisms responsible for biosorption under the presence of divalent cations; however, the former was most significant at low pH, whereas the latter was predominant near neutral pH. The effect of ionic strength on HS biosorption followed the colloidal chemistry theory as the electric double layer became compressed when the ionic strength increased, resulting in closer approach of HS and AS biomass. The humic acid fraction of Suwannee River was removed more efficiently than its fulvic acid fraction because the humic acid was more hydrophobic. These results showed that pH, divalent cation concentration and ionic strength play an important role in the fate and removal of influent wastewater HS in full-scale treatment plants. PMID- 12727239 TI - Pulsed laser irradiation impact on two marine diatoms Skeletonema costatum and Chaetoceros gracilis. AB - The ability of pulsed laser irradiations to cause damage on the biofouling organisms is recently being investigated. If this technique is employed in industries such as power generation wherein a large quantity of water is being used for the cooling purpose, many organisms other than the targeted would get affected. In this study, we have investigated the damage caused by the pulsed laser irradiations from an Nd:YAG laser (fluence 0.1J/cm(2)) for varying durations such as 2, 5, 10, 30, 60 and 300 s on two marine diatom species namely Skeletonema costatum and Chaetoceros gracilis. Upon exposure to low power laser irradiations, these diatom species showed mortalities between 52.6+/-9.3% to 97.7+/-3.1% in the case of S. costatum and 57.8+/-2.5% to 98.9+/-0.6% in the case of C. gracilis for 2 and 300 s of irradiations, respectively. The mortality increased with the increase in the duration of laser irradiation. The estimation of the chlorophyll a concentration in the irradiated samples showed a considerable reduction varying between 9.8% and 57% in C. gracilis and 3% and 70.3% in S. costatum for 2 and 300 s of irradiations, respectively. The laser survived cells grew as the non-irradiated (control) samples. C. gracilis frustules were broken by the laser whilst, the cell materials were drained out of the frustules in the case of S. costatum. The study therefore showed that the low power pulsed laser irradiations could cause significant damage on the two species of planktonic diatoms. PMID- 12727240 TI - Slow and very slow desorption of organic compounds from sediment: influence of sorbate planarity. AB - The kinetics of desorption of in situ chlorobenzenes, PAHs, and PCBs from four different sediments was studied employing Tenax beads as an infinite sink for sorbates. Rate constants for slow desorption were 2.9+/-0.4 x 10(-2) x h(-1), irrespective of the extent of sorbate planarity. Rate constants for very slow desorption were 2.1+/-0.5 x 10(-4) and 6.7+/-1.4 x 10(-4) x h(-1) for planar and non-planar compounds, respectively. Comparison with literature data suggests a priori estimates for rate constants for slow desorption to be 3 x 10(-2) x h(-1), and to be 2 x 10(-4) and 7 x 10(-4) x h(-1) for very slow desorption of planar and non-planar compounds, respectively. The ratio between the fractions in the very slowly desorbing domain and the rapidly desorbing domain was 15-38 for planar compounds which is higher than for non-planar compounds for which the ratio was 2.8-5.2. The ratio between the fractions in the slowly desorbing domain and the rapidly desorbing domain was 1.3-1.8 and independent of the sorbate planarity. The difference in influence of sorbate planarity on the very slowly desorbing domain as compared to the slowly desorbing domain points to different environments for the slowly and the very slowly desorbing fractions. PMID- 12727241 TI - Removal of copper ions from aqueous solution by tree fern. AB - Tree fern, an agricultural by-product, was used for the sorptive removal of copper ions from aqueous solution. The experimental data was analysed by Langmuir, Freundlich and Redlich-Peterson isotherms. The equilibrium sorption capacity of copper ions was determined from the Langmuir equation and found to be 11.7 mg/g. A batch sorption model, based on the assumption of the pseudo-second order mechanism, was developed to predict the rate constant of sorption, the equilibrium sorption capacity and the initial sorption rate with the effect of initial copper ion concentration and the tree fern dose. Various thermodynamic parameters, such as Delta G(0), Delta H(0) and Delta S(0), have been calculated. The thermodynamics of copper ion/tree fern system indicates spontaneous and endothermic nature of the process. PMID- 12727242 TI - Impulse responses of a monoethylamine-fed fluidized bed reactor. AB - The responses of a steady-state, continuous-flow, completely-mixed fluidized bed reactor (FBR) to a range of monoethylamine (MEA) impulses are analyzed in terms of its combined carbon oxidation and nitrification efficiencies. Immobilized cells are cultivated at a mean cell residence time (MCRT) that exceeds 75 days. Responses due to bacterial activities and physical flows are separately estimated using a methodology based on mass balance calculations. MEA inhibition becomes evident when respective critical impulse loadings are exceeded, i.e., 0.12 mg TOC/mg VS for carbon oxidation and 0.021 mg TOC/mg VS for nitrification (TOC: total organic carbon, VS: volatile solids). Nitrifying cells are shown to be more susceptible to MEA impulses than their heterotrophic counterparts. However, the presence of nitrification activities under the conditions tested demonstrates the advantages of cell immobilization that offer greater flexibility when challenged with suddenly increased MEA loads over a short period of time. Mass balance calculations on nitrogen species confirms that 0.583 mg NH(4)(+)-N is produced per mg MEA-C removed when the assimilatory nitrogen requirements for cell synthesis are negligible. PMID- 12727243 TI - Treatment of trichlorophenol by catalytic oxidation process. AB - The oxidation of 2,4,6-trichlorophenol (TCP) by ferrous-catalyzed hydrogen peroxide was quantified and modeled in the study. TCP was effectively degraded by hydroxyl radicals that were generated by Fe(II)/H(2)O(2) in the oxidation process. The oxidation capacity (OC) of the process depends on the concentrations of oxidant (hydrogen peroxide) and oxidative catalyst (ferrous ion). Up to 99.6% of TCP removal can be achieved in the process, provided the doses of Fe(II) and H(2)O(2) are selected correctly. The OC of the process was successfully predicted through a kinetic approach in a two-stage model with some simple and measurable parameters, which makes the model useful for predicting, controlling and optimizing the catalyzed oxidation process in the degradation of TCP. PMID- 12727244 TI - Competitive adsorption of benzoic acid and p-nitrophenol onto activated carbon: isotherm and breakthrough curves. AB - Three series of batch tests at 25 degrees C were performed to determine the benzoic acid and p-nitrophenol (PNP) binary adsorption isotherms onto GAC in the aqueous solutions and the experimental data were fitted to the extended Langmuir isotherm model successfully. The experimental data and the isotherm model parameters showed that the GAC used in this study had a higher affinity to PNP than benzoic acid. Three column tests were performed to determine the breakthrough curves and effluent solution pH with varying feed compositions. According to the experimental results, the weakly adsorbed BA exhibited an intermediate zone of effluent concentration higher than its feed one; the effluent solution pH could serve as a good indicator for breakthrough. The breakthrough curves with varying feed compositions could be predicted by the non linear wave propagation theory satisfactorily. Only the adsorption isotherm models were required to construct the composition path diagram with which the breakthrough curves could be predicted. PMID- 12727245 TI - Use of a mathematical model in the analysis of survival curves of Daphnia magna exposed to toxicants. AB - Chronic toxicity tests carried out on species of the genus Daphnia (Crustacea: Cladocera) are bioassays commonly used in ecotoxicology. Mortality in the cohorts exposed to toxicants can be examined by the analysis of survival curves. The shape of these curves may be very different because of inter-individual heterogeneity: the less rectangular is the shape the more different is the probability of dying of daphnids in the cohort. Aim of this paper is to analyze the characteristics-in particular the shape-of survival curves of cohorts of Daphnia magna exposed to heavy metals in chronic toxicity tests. Experimental curves were fitted by a mathematical model recently proposed, which puts emphasis just on this heterogeneity, described by the parameter of the model S(0). The information contained in S(0) is very useful for the present purposes: the higher the value of S(0) the less rectangular the shape of the curve and, as a consequence, the higher the age-at-death heterogeneity of the cohort is thought to be. The model contains a second parameter, omega, representing the maximum potential ability of the individuals to survive in a specific environment and is related with the maximal life span. The model fitted well the survival curves in most cases and both S(0) and omega showed statistically different values between treatments, useful for comparisons. It was concluded that S(0) provides a quantitative estimation of curve "rectangularization", useful to check different sensitivities to a specific toxicant concentration among daphnids belonging to the same cohort, while omega provides an estimate of maximal life span. PMID- 12727246 TI - Wastewater disinfection with PAA and UV combined treatment: a pilot plant study. AB - This study is part of a larger research project on Advanced Treatments for wastewater reuse in agriculture. Because of Italy's strict microbiological limits on unrestricted wastewater reuse in agriculture (2 MPN/100ml Total Coliforms), a very high degree of disinfection is necessary. The objective of this study is to proceed in validating, with a pilot plant experimentation, previous laboratory results on the disinfection efficacy of the synergic combined treatment between ultraviolet irradiation (UV) and peracetic acid (PAA). The research has been carried out through a 5 month on-site experimental study in a pilot plant, considering four different solutions: PAA addition, UV irradiation, addition of PAA upstream the UV device (PAA+UV) and addition of PAA downstream the UV device (UV+PAA). In the investigated experimental conditions (2-8 ppm of PAA with 10-30 min contact time; 100-300 mJ/cm(2) UV), it has been impossible to meet the microbiological limits through an exclusive use of UV irradiation or PAA. The disinfection efficacy enhances by using the UV+PAA treatment, but a much higher efficacy gain occurs by using the PAA+UV treatment. In this latter case, the higher efficiency is recognized as being brought about by the formation of free radicals due to the photolysis of the PAA when in presence of the UV rays. A preliminary cost analysis has been carried out in order to highlight the more economically advantageous solution which guarantees compliance to the strict limits. PMID- 12727247 TI - Enhanced sonochemical decomposition of 1,4-dioxane by ferrous iron. AB - The enhanced ultrasonic decomposition of 1,4-dioxane by the addition of ferrous iron (Fe(II)) was investigated at 205, 358, 618, and 1071 kHz. The total organic carbon (TOC) remaining was also determined at each frequency. Addition of Fe(II) improved the 1,4-dioxane decomposition rate and mineralization efficiency at all frequencies studied. A nearly four-fold increase of the rate constant was observed at the optimal Fe(II) concentration and a frequency of 205 kHz. In the presence and absence of the iron, the fastest overall degradation and mineralization of 1,4-dioxane took place at 358 kHz where 95% of the initial 1,4 dioxane was removed after 50 min. Finally, although reduced, the ultrasonic decomposition of 1,4-dioxane was still significant at all frequencies in the presence of the hydroxyl radical scavenger bicarbonate. PMID- 12727248 TI - A knowledge-based approach to the deflocculation problem: integrating on-line, off-line, and heuristic information. AB - A knowledge-based approach for the supervision of the deflocculation problem in activated sludge processes was considered and successfully applied to a full scale plant. To do that, a methodology that integrates on-line, off-line and heuristic information has been proposed. This methodology consists of three steps: (i). development of a decision tree (which involves knowledge acquisition and representation); (ii). implementation into a rule-based system; and (iii). validation. The set of symptoms most useful in diagnosing the deflocculation problem has been identified, the different branches to diagnose pin-point floc and dispersed growth have been built (using generic and specific knowledge), and all this knowledge has been codified into an object-oriented shell. The results obtained in the application of this knowledge-based approach to the Granollers WWTP (which treats about 130000 inhabitants-equivalents) showed that the system was able to identify correctly the problem with reasonable accuracy. Our positive experience building this system suggests that this approach is a practical and valuable element to include in an intelligent supervisory system combining numerical and reasoning techniques. PMID- 12727249 TI - Activated sludge exopolymers: separation and identification using size exclusion chromatography and infrared micro-spectroscopy. AB - Extracellular polymeric substances were extracted from activated sludge using a resin exchange method and analyzed. The separation and identification of EPS were carried out by size exclusion chromatography and Fourier transform infrared micro spectroscopy. Chromatograms of extracted EPS exhibited seven peaks. Proteins varying in molecular weights from 670 to 45 kDa were present in all the peaks. Polysaccharides corresponding to molecular weights of approximately 1 and approximately 0.5 kDa were present in only three peaks. Strong association of polysaccharides and proteins was observed. Infrared results revealed the presence of one type of polysaccharide and two types of proteins (A and B). Proteins differed mainly in the length of their associated alkyl chains and in the ratio of ester/acidic functionalities. PMID- 12727250 TI - Denitrification at low temperatures using a suspended carrier biofilm process. AB - The denitrification process was studied in a stirred lab-scale suspended carrier biofilm reactor at low temperatures (3-20 degrees C). The reactor was filled to 50% with Kaldnes K1 carriers. The denitrification rate showed only a rather weak dependence on the temperature, the rate at 3 degrees C being approximately 55% of that at 15 degrees C. The maximum denitrification rate obtained at 15 degrees C was 2.7 g NO(x)(-)-Nm(-2)carrier d (-1). The maximum denitrification rate at 3 degrees C during an 8-day period was found to be constant. During the 8 days, the hydraulic retention time was approximately 1.5h and the inlet NO(3)(-)-N concentration was 30 mg x l(-1). PMID- 12727251 TI - Electro-catalytic oxidation of phenol on several metal-oxide electrodes in aqueous solution. AB - Elecrtochemical degradation of phenol was evaluated at five typical anodes for mineralization to carbon dioxide or for being a pre-treatment method in toxic aromatic compounds. Three kinds of RuO(2)-base electrodes were prepared by thermal deposition, which were coated by the oxides of Ru or by Ru, Sn and Sb or by Ru, Sn, Sb and Gd on Ti metal surface, respectively. Another electrode Ti/ PbO(2) was prepared by electro-deposition method with PbO(2) coated on Ti. A Pt electrode was chosen for comparison. Characteristics of the typical five electrodes were investigated by cyclic voltammetry, SEM and its degradation ability for phenol. Performance for phenol degradation of the three RuO(2) electrodes lie in: Ti/Sb-Sn-RuO(2)-Gd> Ti/Sb-Sn-RuO(2)> Ti/RuO(2) and the electrode with beta-PbO(2) coating was superior to RuO(2)-based electrodes and Pt electrode. Aromatic ring opening take place at all researched electrodes and it is supposed that electrolysis run stop at different intermediates, such as benzoquinone, maleic acid, etc. Under the present experimental conditions, whole mineralization to CO(2) takes place only in the beta-PbO(2) anode. A pathway of electrochemical degradation of phenol was suggested based on the experimental analysis. PMID- 12727252 TI - The use of vegetation to remediate soil freshly contaminated by recalcitrant contaminants. AB - The use of vegetation to remediate soil contaminated by recalcitrant hydrocarbons was tested under field conditions. Specifically, an evaluation was made of the effectiveness of deep rooting grasses, Johnsongrass and Canadian wild rye in the dissipation of TNT and PBB's in the soils freshly contaminated to an initial concentration of 10.17+/-1.35 for TNT and 9.87+/-1.23 mg/kg for PBB. The experiment used 72 (1.5m long and 0.1m diameter) column lysimeters with four treatments: Johnsongrass; wild rye grass; a rotation of Johnsongrass and wild rye grass; and unplanted fallow conditions. In the laboratory, immunoassay test procedures determined the TNT and PBB concentrations in the soil, leachate, herbage and root samples. The root characteristics such as total root length, rooting density, and root surface area were quantified to a depth of 1.5m. Changes in microbial biomass were assessed for both rhizosphere soil and the bulk soil during the 2-year study. The largest and most rapid loss in soil chemical concentration was for TNT, which decreased to less than 250 microg/kg, the detection limit, by 93 days after germination. The PBB was at or near the detection limit of 500 microg/kg by 185 days after germination. There was no perceptible difference in contaminant concentration in the soil between the vegetation treatments and/or with depth. PMID- 12727253 TI - Accumulation of organochlorinated pesticides by triolein-containing semipermeable membrane device (triolein-SPMD) and rainbow trout. AB - An important advance in aquatic ecotoxicology is the development and application of biomimetic sampling technology for hydrophobic contaminants. In this paper, accumulation kinetics of five organochlorinated pesticides, i.e. hexachlorobenzene, lindan, aldrin, heptachlor epoxide and 4,4'-DDT, by triolein containing semipermeable membrane device (triolein-SPMD) and by fish (rainbow trout) were compared in a laboratory continuous flow system. Accumulation kinetics of organochlorinated pesticides by the triolein-SPMD and by rainbow trout were linear during the exposure period of 20 d, except for lindan. Approach of an asymptote could be observed for lindan at the end of the exposure for rainbow trout. When the kinetic data were used to calculate the first-order uptake rate constants, it was found that the uptake rates of the chemicals in triolein-SPMD were 1-2.5 times higher than those in the fish. The estimated bioconcentration factors of the five pesticides in rainbow trout were in the range of 1000-7000 and the device concentration factor in the range of 1500 18000. Although equilibrium was not reached for most pesticides under 21-d exposure period, correlations between C(L)/C(w) and K(ow) (R(2)=0.887) and C(f)/C(w) and K(ow) (R(2)=0.931) could be observed which was a clear indication that triolein-SPMD accumulates chlorinated pesticides in a quite similar way as fish do. Our results show that triolein-SPMD could serve as a good surrogate for rainbow trout for simulating accumulations of chlorinated pesticides and may be used as a universal surrogate for fish in natural waters. PMID- 12727254 TI - Membrane treatment by nanofiltration of exhausted vegetable tannin liquors from the leather industry. AB - A rational use of water in the leather industry is described by applying the criteria of "clean technologies" by means of membrane processes. A nanofiltration (NF) process was used for recovering tannins and water from exhausted baths and reusing them as tanning agents and washings. Results of experiments performed on laboratory and semi-industrial pilot scale plants, operating and fluid-dynamic conditions and mass balance of the NF process are reported and discussed. Chemical and physical analytical values measured on skins treated with the recovered solutions were very similar to those measured on control skins tanned with standard solutions. According to the obtained results, a process scheme is suggested. It permits both to recover water and to increase the tannin/non tannin (T/NT) ratio in the retentate solution, starting from exhausted tanning baths and by using a NF membrane purification/concentration. The final volume reduction factor of tested exhausted baths was 5.25, the increase of T/NT ratio was from 1.1 to 2.3 and the average permeate flux was 12.5l/m(2)h. Advantages are in terms of: reduction of environmental impact (up to 75% of COD reduction of the global effluent), simplification of cleaning-up processes of wastewaters, decrease of disposal costs, saving of chemicals and water. PMID- 12727255 TI - Heterotroph anoxic yield in anoxic aerobic activated sludge systems treating municipal wastewater. AB - As input to the steady state design and kinetic simulation models for the activated sludge system, the correct value for the heterotroph anoxic yield is essential to provide reliable estimates for the system denitrification potential. This paper examines activated sludge anoxic yield values in the literature, and presents experimental data quantifying the value. In the literature, in terms of the structure of ASM1 and similar models, theoretically it has been shown that the anoxic yield should be reduced to approximately 0.79 the value of the aerobic yield. This theoretical value is validated with data from corresponding aerobic OUR and anoxic nitrate time profiles in a batch fed laboratory scale long sludge age activated sludge system treating municipal wastewater. The value also is in close agreement with values in the literature measured with both artificial substrates and municipal wastewater. Thus, it is concluded that, in ASM1 and similar models, for an aerobic yield of 0.67mg COD/mg COD, the anoxic yield should be about 0.53 mg COD/mg COD. Including such a lower anoxic yield in ASM1 and similar models will result in a significant increase in denitrification potential, due to increased denitrification with wastewater RBCOD as substrate. In terms of the structure of ASM3, for the proposed substrate storage yields and the aerobic yield of 0.63 mg COD/mg COD, experimental data indicate that the corresponding anoxic yield should be about 0.42 mg COD/mg COD. This is significantly lower than the proposed value of 0.54 mg COD/mg COD, and requires further investigation. PMID- 12727256 TI - Reactor design and kinetics study of 4,4'-dichlorobiphenyl photodecay in surfactant solution by using a photosensitizer and hydrogen source. AB - The system design based on the photodegradation kinetics of 4,4'-dichlorobiphenyl (4,4'-DCB) in surfactant solution with the aid of solvents (acetone and/or squalane) has been studied. Organic solvents acetone and squalane were added as a photosensitizer and a hydrogen source, respectively, to achieve better photolysis performance. The quantum yield of 4,4'-DCB photodecay in Tween 80 micellar solution in the presence of added acetone was increased from 0.032 to 0.043 at its optimal condition. Acetone was shown to be an effective photosensitizer at low concentration, but an overdose would quench the reaction. Furthermore, the addition of squalane could further promote the photodechlorination of 4,4'-DCB in the Tween 80/acetone solutions for an additional 59% of rate improvement. However, an overdose of either solvent was found to cause UV light attenuation and to reduce the observed quantum yield. This effect has been justified and quantified in this study by a proposed light attenuation model, which has also been incorporated into the kinetic equation so that the resulted formula can be used to design the UV reactors for water and wastewater treatment works. PMID- 12727257 TI - Effect of temperature on removal of heavy metals from contaminated river sediments via bioleaching. AB - The aim of this study was to determine the effect of temperature on the solubilization of heavy metals from contaminated river sediment by sulfur oxidizing bacteria taken from Ell-Ren River sediment. Of three temperatures tested (25 degrees C, 37 degrees C and 55 degrees C), pH decrease was greatest at 37 degrees C, indicating that, after acclimation, bacterial oxidizing activity is greatest at this temperature. At 55 degrees C, pH change was similar to that which occurred with no inoculum added. The increase in sulfates and high pH at 55 degrees C indicate that the indirect mechanism was not initiated at this temperature. Solubilization efficiency of total extractable Ni, Zn, Cu and Cr was high (>90%) at 37 degrees C, whilst that of Pb was only 60.4%. Except for Pb, the optimal temperature for solubilization of total extractable heavy metal was 37 degrees C. The order of average solubilization efficiency of total extractable heavy metals was Ni, Zn, Cu>Cr>Co, Pb. The solubilization efficiency of Pb and Co was markedly less than that of other heavy metals. Transfer of heavy metals between binding fractions was most apparent at 55 degrees C before and after bioleaching. PMID- 12727258 TI - Applications and limitations of the colloid titration method for measuring activated sludge surface charges. AB - Surface charge quantification of polymer solutions and sludge suspensions were investigated by the colloid titration technique and compared to charge densities obtained by pH-titration. The colloid titration technique worked well for polymer samples. The charge quantity of humic acid and activated sludge extracellular polymers (EPS) was estimated to be -1.51 and -0.42 meq/g, respectively. These values are reasonable when compared to pH-titration results. The surface charge of activated sludge particles appears to be below the limit of detection. However, surface charge estimates are obtained, when the reactant doses and sample concentration are increased. It is suggested that such estimates are not correct, but artefacts of the non-stoichiometric precipitation of the polymeric reactants at high doses. It appears that the colloid titration method is limited to conditions of low reactant doses and valid for charge determination of extracted sludge polymers, whereas the method is not valid for charge determination of whole sludge. PMID- 12727259 TI - Selective optimization in thermophilic acidogenesis of cheese-whey wastewater to acetic and butyric acids: partial acidification and methanation. AB - For partial acidogenesis of cheese-whey wastewater, a set of experiments were carried out to produce short-chain volatile fatty acids (VFA) in laboratory-scale continuously stirred tank reactors (CSTR). The maximum rate of acetic and butyric acid production associated with simultaneous changes in hydraulic retention time (HRT), pH, and temperature was investigated, in which the degree of acidification of the whey to the short-chain VFAs was less than 20% of the influent chemical oxygen demand (COD) concentration. Response surface methodology was successfully applied to determine the optimum physiological conditions where the maximum rates of acetic and butyric acid production occurred. These were 0.40-day HRT, pH 6.0 at 54.1 degrees C and 0.22-day HRT, pH 6.5 at 51.9 degrees C, respectively. The optimum conditions for acetic acid production were selected for partial acidification of cheese-whey wastewater because of a higher rate in combined productions of acetic and butyric acids than that at optimum conditions for butyric acid production. A thermophilic two-phase process with the partial acidification followed by a methanation step was operated. Performance of the two phase process was compared to the single-phase anaerobic system. The two-phase process clearly showed a better performance in management of cheese-whey wastewater over the single-phase system. Maximum rate of COD removal and the rate of methane production in the two-phase process were, respectively, 116% and 43% higher than those of the single-phase system. PMID- 12727260 TI - Arsenic removal using steel manufacturing byproducts as permeable reactive materials in mine tailing containment systems. AB - Steel manufacturing byproducts were tested as a means of treating mine tailing leachate with a high As concentration. Byproduct materials can be placed in situ as permeable reactive barriers to control the subsurface release of leachate from tailing containment systems. The tested materials had various compositions of elemental Fe, Fe oxides, Ca-Fe oxides and Ca hydroxides typical of different steel manufacturing processes. Among these materials, evaporation cooler dust (ECD), oxygen gas sludge (OGS), basic oxygen furnace slag (BOFS) and to a lesser degree, electrostatic precipitator dust (EPD) effectively removed both As(V) and As(III) during batch experiments. ECD, OGS and BOFS reduced As concentrations to <0.5mg/l from 25mg/l As(V) or As(III) solution in 72 h, exhibiting higher removal capacities than zero-valent iron. High Ca concentrations and alkaline conditions (pH ca. 12) provided by the dissolution of Ca hydroxides may promote the formation of stable, sparingly soluble Ca-As compounds. When initial pH conditions were adjusted to 4, As reduction was enhanced, probably by adsorption onto iron oxides. The elution rate of retained As from OGS and ECD decreased with treatment time, and increasing the residence time in a permeable barrier strategy would be beneficial for the immobilization of As. When applied to real tailing leachate, ECD was found to be the most efficient barrier material to increase pH and to remove As and dissolved metals. PMID- 12727261 TI - Appropriate conditions or maximizing catalytic reduction efficiency of nitrate into nitrogen gas in groundwater. AB - This study focused on the appropriate catalyst preparation and operating conditions for maximizing catalytic reduction efficiency of nitrate into nitrogen gas from groundwater. Batch experiments were conducted with prepared Pd and/or Cu catalysts with hydrogen gas supplied under specific operating conditions. It has been found that Pd-Cu combined catalysts prepared at a mass ratio of 4:1 can maximize the nitrate reduction into nitrogen gas. With an increase in the quantity of the catalysts, both nitrite intermediates and ammonia can be kept at a low level. It has also been found that the catalytic activity is mainly affected by the mass ratio of hydrogen gas to nitrate nitrogen, and hydrogen gas gauge pressure. Appropriate operating values of H(2)/NO(3)-N ratio, hydrogen gas gauge pressure, pH, and initial nitrate concentration have been determined to be 44.6g H(2)/g N, 0.15 atm, 5.2 (-), 100 mg x L(-1) for maximizing the catalytic reduction of nitrate from groundwater. PMID- 12727262 TI - Degradation of monomethylmercury chloride by hydroxyl radicals in simulated natural waters. AB - The degradation of methylmercury chloride by hydroxyl radicals (*OH) has been investigated using nitrate photolysis from 285 to 800 nm with a 450 W Xenon lamp as the (*OH source. The identified products are Hg(2+), Hg(0), CHCl(3) and formaldehyde. The second-order rate constant at pH of 5 at room temperature was determined to be 9.83(+-0.66)x10(9) M(-1) x s(-1)using benzoic acid as the *OH scavenger. The effects of chloride concentration and methylmercury speciation have also been investigated. A mechanism of the CH(3)HgCl-*OH reaction has been proposed. The calculated methylmercury degradation rates in natural waters using the above rate constant were comparable to the in situ photodegradation rates reported previously, indicating that degradation by (*)OH may be one of the important pathways of methylmercury degradation in sunlit surface waters. PMID- 12727263 TI - Bioaccumulation of nickel from aqueous solutions by genetically engineered Escherichia coli. AB - This study constructed a genetically engineered Escherichia coli JM109 which simultaneously expressed nickel transport system and metallothionein to remove and recover Ni(2+) from aqueous solution. Bioaccumulation process was rapid and followed linearized Langmuir isotherm. A more than six-fold increase of Ni(2+) binding capacity was obtained by genetically engineered E. coli cells compared with original host E. coli cells. A pH assay showed genetically engineered E. coli cells accumulated Ni(2+) effectively over a broad range of pH (4-10). The presence of 1000 mg/L Na(+) and Ca(2+), or 50mg/L Cd(2+) or Pb(2+) did not have a significant effect on Ni(2+) bioaccumulation, while Mg(2+), Hg(2+) and Cu(2+) posed a severe adverse influence on Ni(2+) uptake by genetically engineered E. coli. Furthermore, genetically engineered E. coli cells did not require extra nutrients for Ni(2+) bioaccumulation. PMID- 12727264 TI - Electrocatalytic reduction of nitrate in water. AB - Nitrate (NO(3)(-)) contamination of groundwater is a common problem throughout intensive agricultural areas (nonpoint source pollution). Current processes (e.g., ion exchange, membrane separation) for NO(3)(-) removal have various disadvantages. The objective of this study was to evaluate an electrocatalytic reduction process to selectively remove NO(3)(-) from groundwater associated with small agricultural communities. A commercially available ELAT (E-Tek Inc., Natick, MA) carbon cloth with a 30% surface coated Rh (rhodium) (1microg x cm( 1)) was tested at an applied potential of -1.5 V versus standard calomel electrode (SCE) with a Pt auxiliary electrode. Electrocatalytic reduction process (electrolysis) of NO(3)(-) was tested with cyclic voltammetry (CV) in samples containing NO(3)(-) and 0.1M NaClO(4)(-). Nitrate and NO(2)(-) concentrations in test solutions and groundwater samples were analyzed by ion chromatography (IC). The presence of Rh on the carbon cloth surface resulted in current increase of 36% over uncoated carbon cloths. The electrocatalysis experiments using Rh coated carbon cloth resulted in reduction of NO(3)(-) and NO(2)(-) on a timescale of minutes. Nitrite is produced as a product, but is rapidly consumed upon further electrolysis. Field groundwater samples subjected to electrocatalysis experiments, without the addition of NaClO(4)(-) electrolyte, also exhibited removal of NO(3)(-) on a timescale of minutes. Overall, results suggest that at an applied potential of -1.5 V with respect to SCE, Rh coated carbon cloth can reduce NO(3)(-) concentrations in field groundwater samples from 73 to 39 mg/L (16.58 to 8.82 mg/L as N) on a timescale range of 40-60 min. The electrocatalytic reduction process described in this study may prove useful for removing NO(3)(-) and NO(2)(-) from groundwater associated with nonpoint source pollution. PMID- 12727265 TI - Wastewater treatment by radial freezing with stirring effects. AB - Radial freezing experiments on wastewater models were conducted in the presence of imposed stirring in order to remove impurities. The studied samples (dilute Na montmorillonite suspensions charged with nitrates and with zinc or lead) were placed inside a cylindrical annulus, cooled at a controlled temperature around -7 degrees C at its inner wall which rotated around a vertical axis. The freezing front propagated toward the still outer wall which was maintained at a constant temperature around +1 degrees C. Thanks to stirring, considerable purification rates up to 99.97% were attained. It was also demonstrated that combining radial freezing and stirring ended in residual concentrations which agreed with drinking water standards. PMID- 12727266 TI - A microscopic system with a dual band filter for the simultaneous enumeration of Cryptosporidium parvum oocysts and sporozoites. PMID- 12727268 TI - Purification and characterization of Hainantoxin-V, a tetrodotoxin-sensitive sodium channel inhibitor from the venom of the spider Selenocosmia hainana. AB - A neurotoxic peptide, named Hainantoxin-V (HNTX-V), was isolated from the venom of the Chinese bird spider Selenocosmia hainana. The complete amino acid sequence of HNTX-V has been determined by Edman degradation and found to contain 35 amino acid residues with three disulfide bonds. Under whole-cell patch-clamp mode, HNTX V was proved to inhibit the tetrodotoxin-sensitive (TTX-S) sodium currents while it had no any effects on tetrodotoxin-resistant (TTX-R) sodium currents on adult rat dorsal root ganglion neurons. The inhibition of TTX-S sodium currents by HNTX V was tested to be concentrate-dependent with the IC(50) value of 42.3nM. It did not affect the activation and inactivation kinetics of currents and did not have the effect on the active threshold of sodium channels and the voltage of peak inward currents. However, 100nM HNTX-V caused a 7.7mV hyperpolarizing shift in the voltage midpoint of steady-state sodium channel inactivation. The results indicated that HNTX-V inhibited mammalian voltage-gated sodium channels through a novel mechanism distinct from other spider toxins such as delta-ACTXs, micro agatoxins I-VI which bind to receptor site three to slow the inactivation kinetics of sodium currents. PMID- 12727269 TI - Purification and characterization of raventoxin-I and raventoxin-III, two neurotoxic peptides from the venom of the spider Macrothele raveni. AB - The spider Macrothele raveni was recently identified as a new species of Genus Macrothele. The crude venom from M. raveni was found to be neurotoxic to mice and the LD(50) of the crude venom in mice was 2.852mg/kg. Two neurotoxic peptides, raventoxin-I and raventoxin-III, were isolated from the crude venom by ion exchange and reverse phase high performance liquid chromatography. Raventoxin-I was the most abundant toxic component in the venom, while raventoxin-III was a lower abundant component. Both toxins can kill mice and block neuromuscular transmission in an isolated mouse phrenic nerve diaphragm preparation, but have no effect on cockroaches. The LD(50) of raventoxin-I in mice is 0.772mg/kg. The complete amino acid sequences of raventoxin-I and raventoxin-III were determined and found to consist of 43 and 29 amino acid residues, respectively. It was determined by mass spectrometry that all Cys residues from raventoxin-I and raventoxin-III are involved in disulphide bonds. raventoxin-III showed no significant sequence homology with any presently known neurotoxins in the protein/DNA databases, while raventoxin-I has limited sequence identity with delta-AcTx-Hv1 and delta-AcTx-Ar1, which target both mammalian and insect sodium channels. Both raventoxin-I and raventoxin-III only work on vertebrates, but not on insects. Moreover, raventoxin-I could exert an effect of first exciting and then inhibiting the contraction of mouse diaphragm muscle caused by electrically stimulating the phrenic nerve, but raventoxin-III could not. PMID- 12727270 TI - Neurotoxic and myotoxic actions of Naja naja kaouthia venom on skeletal muscle in vitro. AB - The neuromuscular and skeletal muscle actions of Naja naja kaouthia snake venom were studied in mammalian (rat left hemidiaphragm) and avian (chick biventer cervicis) nerve-muscle preparations. The venom (5 and 10 micro g/ml) produced neuromuscular blockade (85% in 36.8+/-2.0min, mean+/-SEM, n=5, and 18+/-0.6min, n=3, p<0.01, respectively) in the rat preparation. That the phospholipase A(2) (PLA(2)) activity of the venom is involved in this effect was evaluated by inhibiting this enzyme with p-bromophenacyl bromide. This resulted in significantly (p<0.01) increasing the time required for 85% blockade with 5 and 10 micro g/ml to 54+/-4.6min (n=3) and 29+/-0.6min (n=3), respectively. In chick preparations, the venom (5 micro g/ml) produced neuromuscular blockade in 14.0+/ 1.8min (n=5). The contractures to exogenous acetylcholine were completely inhibited by the venom, whereas those to 134 micro M KCl were partially blocked in chick preparations (n=4, n=3, respectively). The venom (5 micro g/ml) produced a progressive decrease in the amplitude of miniature end-plate potentials (m.e.p.ps) in the rat hemidiaphragm, but did not alter the resting membrane potential at 5 micro g/ml. Neostigmine (5.8 micro M) immediate and partially reversed the 85% blockade produced by venom (61%, n=3) in rat preparations, as did 4-aminopyridine (53 micro M) ( approximately 59%, n=3). The 4-aminopyridine and neostigmine also restored the m.e.p.ps to pre-venom (control) values. In rat preparations, the venom damaged 47%+/-11% and 62.7+/-3.6% of the muscle fibers at concentrations of 5 and 10 micro g/ml, respectively. For venom in which PLA(2) activity was inhibited, the corresponding values were 38+/-11.8% (5 micro g/ml) and 67+/-9.6% (10 micro g/ml). These findings suggest a post-synaptic neurotoxic action for N. n. kaouthia venom, and that inhibiting phospholipase activity of the venom reduces significantly the neuromuscular block but not the direct myotoxicity. PMID- 12727271 TI - Hyperalgesia induced by Asp49 and Lys49 phospholipases A2 from Bothrops asper snake venom: pharmacological mediation and molecular determinants. AB - The ability of Lys49 and Asp49 phospholipases A(2) (PLA(2)), from Bothrops asper snake venom, to cause hyperalgesia was investigated in rats, using the paw pressure test. Intraplantar injection of both toxins (5-20 micro g/paw) caused hyperalgesia, which peaked 1h after injections. Incubation of both proteins with heparin, prior to their injection, partially reduced this response. Chemical modification of Asp49 PLA(2) with p-bromophenacyl bromide (p-BPB), which abrogates its PLA(2) activity, also abolished hyperalgesia. Intraplantar injection of a synthetic peptide corresponding to the C-terminal sequence 115-129 of Lys49 PLA(2), caused hyperalgesia of similar time course, but varying magnitude, than that induced by the native protein. In contrast, a homologous peptide derived from the Asp49 PLA(2) did not show any nociceptive effect. Hyperalgesia induced by both PLA(2)s was blocked by the histamine and serotonin receptor antagonists promethazine and methysergide, respectively, by the bradykinin B(2) receptor antagonist HOE 140 and by antibodies to tumor necrosis factor alfa (TNFalpha) and interleukin 1 (IL-1). Pretreatment with guanethidine, atenolol, prazosin and yohimbine, inhibitors of sympathomimetic amines, or with indomethacin, inhibitor of the cyclo-oxygenase pathway, reduced Lys49 PLA(2) induced hyperalgesia without interfering with the nociceptive activity of Asp49 PLA(2). The hyperalgesic response to both myotoxins was not modified by pretreatment with celecoxib, an inhibitor of the cyclo-oxygenase type II, by zileuton, an inhibitor of the lipoxygenase pathway or by N(g)-methyl-L-arginine (LNMMA), an inhibitor of nitric oxide synthase. These results suggest that Asp49 and Lys49 PLA(2)s are important hyperalgesic components of B. asper venom, and that Lys49 and Asp49 PLA(2)s exert their algogenic actions through different molecular mechanisms. PMID- 12727272 TI - Modulation of intracellular Ca2+ levels by Scorpaenidae venoms. AB - The crude venoms of the soldierfish (Gymnapistes marmoratus), the lionfish (Pterois volitans) and the stonefish (Synanceia trachynis) display pronounced neuromuscular activity. Since [Ca(2+)](i) is a key regulator in many aspects of neuromuscular function we sought to determine its involvement in the neuromuscular actions of the venoms. In the chick biventer cervicis muscle, all three venoms produced a sustained contraction (approx 20-30% of 1mM acetylcholine). Blockade of nicotinic receptors with tubocurarine (10 micro M) failed to attenuate the contractile response to either G. marmoratus venom or P. volitans venom, but produced slight inhibition of the response to S. trachynis venom. All three venoms produced a rise in intracellular Ca(2+) (approx. 200-300% of basal) in cultured murine cortical neurons. The Ca(2+)-channel blockers omega conotoxin MVIIC, omega-conotoxin GVIA, omega-agatoxin IVa and nifedipine (each at 1 micro M) potentiated the increase in [Ca(2+)](i) in response to G. marmoratus venom and P. volitans venom, while attenuating the response to S. trachynis venom. Removal of extracellular Ca(2+), replacement of Ca(2+) with La(3+) (0.5mM), or addition of stonefish antivenom (3units/ml) inhibited both the venom induced increase in [Ca(2+)](i) in cultured neurones and contraction in chick biventer cervicis muscle. Venom-induced increases in [Ca(2+)](i) correlated with an increased cell death of cultured neurones as measured using propidium iodide (1 micro g/ml). Morphological analysis revealed cellular swelling and neurite loss consistent with necrosis. These data indicate that the effects of all three venoms are due in part to an increase in intracellular Ca(2+), possibly via the formation of pores in the cellular membrane which, under certain conditions, can lead to necrosis. PMID- 12727273 TI - Expression, purification, and efficacy of the type A botulinum neurotoxin catalytic domain fused to two translocation domain variants. AB - Clostridial neurotoxins are potent inhibitors of synaptic function, with the zinc dependent proteolytic light chain (LC) portion of the toxin cleaving one of three neural SNARE proteins. In nature, the LC is expressed as a part of a much larger toxin and hemagglutinin complex, protecting it from environmental degradation and preserving its catalytic activity. We developed forms of the LC of type A botulinum neurotoxin (BoNT-A) with parts of the larger toxin gene, for use as reagents in high-throughput assays to screen for potential LC antagonists, to further elucidate the toxin's mechanism of action, and to study immunological responses to the toxin. Three BoNT-A constructs were engineered and expressed: the LC, LC with translocation region (LC+H(n)), and the LC with the belt portion of the translocation region (LC+Belt). Purification was optimized to a two-step process, with relatively high yields of all three constructs obtained. Activity assays showed all three constructs to be active, with the LC being the most active. Immunogenic protection against native BoNT-A toxin challenge was observed for all three constructs, with the best protection observed with the LC+H(n) and LC+Belt proteins. PMID- 12727274 TI - The in vitro effects of two chirodropid (Chironex fleckeri and Chiropsalmus sp.) venoms: efficacy of box jellyfish antivenom. AB - The pharmacological and biochemical isolation of cnidarian venoms has been hindered by difficulties with both extracting pure venom from nematocysts and venom stability. The development of a new technique to extract active, pure venom of Chironex fleckeri and Chiropsalmus sp. has enabled identify both neurotoxic and myotoxic activity in their venoms. These activities are similar, but not identical in each species. Venom (50 micro g/ml) from both species significantly inhibited indirect and direct twitches of the chick biventer nerve-muscle preparation. Pre-incubation with 1U/ml box jellyfish antivenom did not have any significant effect on venom-induced reductions of indirect twitches. However, this activity was markedly attenuated by prior addition of 5U/ml antivenom, albeit to a lesser degree for Chiropsalmus sp. In contrast, prior addition of 5U/ml box jellyfish antivenom did not neutralise the myotoxic activity of C. fleckeri venom (50 micro g/ml), although it did inhibit the myotoxicity produced by Chiropsalmus sp. venom (50 micro g/ml). Antivenom (5U/ml) added 1h after the addition of C. fleckeri venom (50 micro g/ml) had no effect on the indirect or direct twitches of the skeletal muscle preparation. However, it partially restored the reduction in indirect twitch height caused by Chiropsalmus sp. venom (50 micro g/ml). Myotoxicity was confirmed in muscle preparations stained with hematoxylin and eosin.Therefore, although antivenom was able to neutralize the neurotoxic effects of both species, and the myotoxic effects of Chiropsalmus sp., when added prior to venom, it was unable to reverse the effects after venom addition. This suggests that antivenom is unlikely to be useful in the treatment of neurotoxic or myotoxic effects in patients, although these effects are rarely seen clinically. PMID- 12727275 TI - Functional expression and characterization of a recombinant phospholipase A2 from sea snake Lapemis hardwickii as a soluble protein in E. coli. AB - Three full-length phospholipase A(2) (PLA(2)) cDNAs from sea snake Lapemis hardwickii venom were cloned and sequenced in our previous study. In order to investigate their biological functions, we established a fusion expression system for PLA(2)-9 in E. coli. The open reading frame encoding mature peptide of PLA(2) 9 was subcloned into the vector pTRX. The Trx-PLA(2)-9 fusion protein was expressed as a soluble protein by IPTG induction at 23 degrees C. The fusion protein was purified with metal-chelate affinity chromatography and then cleaved by enterokinase. The mature recombinant PLA(2)-9 was further purified by ion exchange chromatography and a final yield of approximately 2.5mg pure PLA(2)-9 from 1l of bacteria culture was obtained. The catalytic activity of recombinant PLA(2)-9 (rPLA(2)-9) was measured and found to be similar to native enzyme. As the Austrelaps superbus PLA(2), which shares 90% nucleotide sequence similarity to PLA(2)-9, the rPLA(2)-9 displayed the anti-platelet aggregation effect. Site directed mutagenesis of the two conserved residues, His-48 and Asp-49, resulted in the loss of catalytic activity, however did not affect the inhibition effect of platelet aggregation suggesting that these two activities of sea snake PLA(2) 9 may be dissociated. PMID- 12727277 TI - Bibliography of toxinology. PMID- 12727281 TI - XVII Helsinki University Congress of Drug Research. June 5-6, 2003 Helsinki, Finland. Abstracts. PMID- 12727276 TI - Cytotoxic and cytostatic activity of erlangerins from Commiphora erlangeriana. AB - The resin of Commiphora erlangeriana is known to be poisonous to humans and animals and has traditionally been used as an arrow poison. Since recent phytochemical studies on this plant material has identified four major lignans (named, Erlangerins A to D) that closely relate to the structure of podophyllotoxin, it was hypothesised that the well known poisoning effect of the resin could in part be due to direct toxicity to mammalian cells. Hence, the toxicity of Erlangerins was studied by measuring the viability of two human (HeLa and EAhy926) and two murine (L929 and RAW 264.7) cell lines. As assessed by the MTT assay, the effect of Erlangerin C and D closely follow the activity profile of podophyllotoxin: they induced a concentration-dependent cytotoxicity in the murine macrophage cells (RAW 264.7) and a cytostatic effect in HeLa, EAhy926 and L929 cells. In contrast, Erlangerins A and B suppressed cell viability at relatively higher concentrations (EC(50) values higher than 3 micro M as compared with nM concentration range for Erlangerins C and D and podophyllotoxin) and their activity appears to be consistent with a cytotoxic mode of action in all cell lines studied. The structure-activity-relationship established from the study is discussed. PMID- 12727282 TI - Kinetic analysis of artificial peptide self-replication. Part I: the homochiral case. AB - Computational kinetic analysis of a lately discovered homochiral peptide self replicator is presented. A 6-step kinetic model was designed that addresses the main reactions and hydrophobic interactions involved in this template-directed, autocatalytic system and that gave rise to excellent fitting of 4 previously published independent experimental series. The model sheds light on the mechanistic principle of the reaction system and illustrates directly a number of dynamic properties such as the observed autocatalytic efficiency. It was found that the dynamics are basically governed by two reversible hydrophobic interactions: between the template and a peptide fragment and between two template species. The later association was determined to be considerably more favored, which leads to the predominant presence of the catalytically inactive template dimer in the reaction system. Our results show that the involvement of a template trimer is not necessary to obtain the observed fittings. PMID- 12727283 TI - Kinetic analysis of artificial peptide self-replication. Part II: the heterochiral case. AB - A kinetic model has been designed to describe and to analyze the stereoselective behavior of a recently discovered heterochiral template-directed peptide self replicator by Ghadiri and co-workers [Nature 409 (2001) 797-801]. It turned out that previous assumptions stating that exclusively homochiral species participate in a stereoselective and autocatalytic pathway and that heterochiral species originate only from uncatalyzed background reactions could not be validated by our model. On the contrary, excellent fitting of experimental data indicated that the whole combinatorial variety of possible cross-catalytic processes involving L and D- peptide species play an important role and need to be taken into account. The system shows no net creation of chiral matter but only a redistribution of the initially present chiral material. Both, the separation of an optically inactive meso-type template dimer and a slight chiroselective autocatalytic effect, contribute to a predicted amplification of enantiomeric excess that, in some cases, can simultaneously result in a substantial amount of optically active matter. PMID- 12727284 TI - Correlation between fluidising effects on phospholipid membranes and mitochondrial respiration of propofol and p-nitrosophenol homologues. AB - Nitrosopropofol (2-6-diisopropyl-4-nitrosophenol) has dramatic consequences for respiration, ATP synthesis and the transmembrane potential of isolated rat liver mitochondria at concentrations at which propofol (2-6-diisopropylphenol) does not cause any apparent effects. These results correlate well with the observation that nitrosopropofol is also a stronger perturbing agent of phospholipid membranes. In this paper we verify the possible biological activity of different phenols and nitrosophenols on mitochondrial respiration. We then discuss their interactions with phospholipid liposomes, studied with differential scanning calorimetry, spin labelling techniques and UV-Vis spectrophotometry, in order to obtain information on drug distribution and the modifications they impose on lipid bilayer. The results of the experiments performed on mitochondria and model membranes prove an interesting correlation between the effects of the molecules on both systems. PMID- 12727285 TI - Inter-subunit recognition and manifestation of segmental mobility in Escherichia coli RNA polymerase: a case study with omega-beta' interaction. AB - Omega (omega), consisting of 91 amino acids, is the smallest of all the Escherichia coli RNA polymerase subunits and is organized into an N-terminal domain of 53 amino acids followed by an unstructured tail in the C-terminal region. Our earlier experiments have shown a chaperone-like function of omega in which it helps to maintain beta' in a correct conformation and recruit it to the alpha(2)beta subassembly to form a functional core enzyme (alpha(2)betabeta'omega). The X-ray structure analysis of Thermus aquaticus core RNA polymerase suggests that two regions of omega latch onto the N-terminal and C terminal ends of the beta'-subunit. In the present study we have monitored the conformational changes in beta' as the denatured protein is refolded in the presence and absence of omega using tryptophan fluorescence emission of beta' as well as acrylamide quenching of Trp fluorescence. Results indicate that the presence of stoichiometric amounts of omega is helpful in beta' refolding. We have also monitored the behavior of the C-terminal tail of omega by engineering three cysteine residues at three different sites in omega and subsequently labeling them with a sulphydryl-specific fluorescent probe. Fluorescence anisotropy measurements of the labeled protein indicate that the C-terminal domain of omega is mobile in the free protein and gets restrained in the presence of beta'. Calculations on side-chain interactions show that out of the three mutated positions, two have near neighbourhood interactions only with side-chains in the beta' subunit whereas the end of the C-terminal of omega, although it is restrained in the presence of beta', has no interacting partner within a 4-A radius. PMID- 12727286 TI - Cytochrome c location in phosphatidylcholine/cardiolipin model membranes: resonance energy transfer study. AB - Resonance energy transfer between lipid-bound fluorescent probe 3 methoxybenzanthrone as a donor and heme group of cytochrome c as an acceptor has been examined to ascertain the protein disposition relative to the surface of model membranes composed of phosphatidylcholine and cardiolipin (10, 50 and 80 mol%). The model of energy transfer in membrane systems has been extended to the case of donors distributed between the two-bilayer leaflets and acceptors located at the outer monolayer taking into account the donor and acceptor orientational behavior. Assuming specific protein orientation relative to the membrane surface and varying lateral distance of the donor-acceptor closest approach in the range from 0 to 3.5 nm the limits for possible heme distances from the bilayer midplane have been found to be 0.8-3 nm (10 mol% CL), 0-2.6 nm (50 mol% CL), and 1.4-3.3 nm (80 mol% CL). PMID- 12727287 TI - Influence of cation-pi interactions in different folding types of membrane proteins. AB - Cation-pi interactions play an important role to the stability of protein structures. In this work, we analyze the influence of cation-pi interactions in three-dimensional structures of membrane proteins. We found that transmembrane strand (TMS) proteins have more number of cation-pi interactions than transmembrane helical (TMH) proteins. In TMH proteins, both the positively charged residues Lys and Arg equally experience favorable cation-pi interactions whereas in TMS proteins, Arg is more likely than Lys to be in such interactions. There is no relationship between number of cation-pi interactions and number of residues in TMH proteins whereas a good correlation was observed in TMS proteins. The average cation-pi interaction energy for TMH proteins is -16 kcal/mol and that for TMS proteins is -27 kcal/mol. The pair-wise cation-pi interaction energy between aromatic and positively charged residues showed that Lys-Trp energy is stronger in TMS proteins than TMH proteins; Arg-Phe, Arg-Tyr and Lys-Phe have higher energy in TMH proteins than TMS proteins. The decomposition of energies into electrostatic and van der Waals revealed that the contribution from electrostatic energy is twice as that from van der Waals energy in both TMH and TMS proteins. The results obtained in the present study would be helpful to understand the contribution of cation-pi interactions to the stability of membrane proteins. PMID- 12727288 TI - Thermodynamic non-ideality as an alternative source of the effect of sucrose on the thrombin-catalyzed hydrolysis of peptide p-nitroanilide substrates. AB - The inhibitory effect of sucrose on the kinetics of thrombin-catalyzed hydrolysis of the chromogenic substrate S-2238 (D-phenylalanyl-pipecolyl-arginoyl-p nitroanilide) is re-examined as a possible consequence of thermodynamic non ideality-an inhibition originally attributed to the increased viscosity of reaction mixtures. However, those published results may also be rationalized in terms of the suppression of a substrate-induced isomerization of thrombin to a slightly more expanded (or more asymmetric) transition state prior to the irreversible kinetic steps that lead to substrate hydrolysis. This reinterpretation of the kinetic results solely in terms of molecular crowding does not signify the lack of an effect of viscosity on any reaction step(s) subject to diffusion control. Instead, it highlights the need for development of analytical procedures that can accommodate the concomitant operation of thermodynamic non-ideality and viscosity effects. PMID- 12727289 TI - Comparison of the interaction, positioning, structure induction and membrane perturbation of cell-penetrating peptides and non-translocating variants with phospholipid vesicles. AB - Cell-penetrating peptides (CPPs) are able to translocate and carry cargo molecules across cell membranes. Using fluorescence techniques (polarization and quenching) and CD spectroscopy we studied the interaction, conformation and topology of two such peptides, transportan and 'penetratin' (pAntp), and two variants of differing translocating abilities, with small phospholipid vesicles of varying charge density. The induced structure of transportan is always helical independent of vesicle surface charge. pAntp and its two variants interact significantly only with negatively charged vesicles. The induced secondary structure depends on membrane charge and lipid/peptide ratio. The degree of membrane perturbation, evidenced by fluorescence polarization, of pAntp and its variants is related to their secondary structure. In the helical state, the peptides have little effect on the membrane. Under conditions where pAntp and its variants are converted into beta-structures, they cause membrane perturbation. Oriented CD suggests that the two CPPs (pAntp and transportan) in their helical state lie along the vesicle surface, while the two pAntp variants appear to penetrate deeper into the membrane. PMID- 12727290 TI - NMR and parity violation: low-temperature dependence in 1H CRAMPS and 13C CP/MAS ssNMR spectra of alanine enantiomer. AB - Life is based on L-amino acids and D-sugars rather than the enantiomeric D-amino acids and L-sugars. This broken symmetry is now believed to be a feature of fundamental physics-a result of symmetry-breaking induced by the weak force, which makes one enantiomer slightly more stable than the other. An amplification mechanism based on quantum mechanical tunneling could give rise to a second-order phase transition. In order to understand the transition mechanism, we measured the temperature dependence of 1H CRAMPS solid state NMR and 13C CP/MAS spectra of D- and L-alanine crystals from 295 K through to 220 K. Obvious difference of NMR behaviors between two enantiomers was observed in the phase transition which may be related to one suggested by Salam, caused biochirality among twenty amino acids. PMID- 12727291 TI - Antiproliferative and apoptotic effect of TGF-beta 1 in bovine mammary epithelial BME-UV1 cells. AB - Transforming growth factor beta1 (TGF-beta(1)) is regarded as an important auto/paracrine regulator of mammary gland involution, however, its apoptotic effect and inhibition of growth in bovine mammary epithelial cells (MEC) has not been documented. In the present study, laser scanning cytometry, confocal and immunoelectron microscopy techniques were used for quantitative and qualitative analyzes of apoptosis, cell cycle and expression, subcellular redistribution and interactions of apoptosis-related proteins in bovine BME-UV1 MEC exposed to TGF beta(1). TGF-beta(1) exerted both antiproliferative and apoptotic action. The antiproliferative effect was manifested by increase of cell number in G1 phase with simultaneous decrease of cell number in S and G2/M phases. It resulted in significant increase of G1/S ratio in TGF-beta(1) treated cells, indicating partial cell cycle arrest at the G1-S transition. Apoptosis induced by TGF beta(1) manifested by characteristic morphological changes. Among biochemical features of TGF-beta(1)-induced apoptosis in BME-UV1 cells we found: (1) an increase of cell number with lowered DNA content and condensed chromatin, (2) enhanced expression of caspase-3 and m-calpain, (3) elevated number of 89 kDa PARP degradation fragments, and (4) aggregation of Bax and its interactions with voltage dependant anion channel-1. In conclusion, antiproliferative and apoptotic action of TGF-beta(1), observed in the culture of BME-UV1 cells, suggests an essential role of this cytokine in the regulation of mammary gland involution in cow. PMID- 12727293 TI - Roles of mu-calpain in cultured L8 muscle cells: application of a skeletal muscle specific gene expression system. AB - The goal of this work was to characterize the roles of mu-calpain in skeletal muscle protein degradation. Three approaches were developed to alter mu-calpain activity in rat myotubes. These included over-expression of antisense mu-calpain (mu-AS), dominant negative mu-calpain (mu-DN) and the antisense 30-kDa calpain subunit (30-AS). Constructs were expressed in rat L8 myotubes, and their effects on protein degradation and on concentrations of intact and/or degraded fodrin, desmin and tropomyosin were examined. An ecdysone-inducible expression system, in which we replaced a constitutively active CMV promoter with a skeletal muscle specific alpha-actin promoter, was used to drive expression. Cell lines were evaluated by expression of the gene-of-interest following addition of ponasterone A (PA; ecdysone analog) to culture medium. Changes in calpain activity were assessed by evaluating fodrin degradation. 30-AS, which should alter both mu- and m-calpain activities, increased intact fodrin concentration. mu-DN and mu-AS reduced fodrin degradation products. mu-DN reduced total protein degradation by 7.9% (P<0.01) at 24 h and by 10.6% (P<0.01) at 48 h. mu-AS reduced total protein degradation by 6.4% at 24 h (P<0.05). 30-AS reduced total protein degradation by 13.4% (P<0.05) and 7.3% (P<0.05) following 24 and 48 h of PA administration, respectively. We assessed effects of mu-DN, mu-AS and 30-AS on concentrations of desmin and tropomyosin. Inhibition of calpains stabilized desmin, but had no effect on tropomyosin. These data indicate that fodrin and desmin are mu-calpain substrates and that mu-calpain accounts for a small proportion of total protein degradation in muscle cells. Tropomyosin is not degraded by calpain in muscle cells. PMID- 12727292 TI - Purification and characterization of alpha 2-macroglobulin from the white shrimp (Penaeus vannamei). AB - alpha(2)-Macroglobulin (alpha(2)M) is a broad-spectrum protease-binding protein abundant in plasma from vertebrates and several invertebrate phyla. This protein was purified from cell-free hemolymph of the white shrimp, Penaeus vannamei, using Blue-Sepharose and Phenyl-Sepharose chromatography. The shrimp alpha(2)M is a 380 kDa protein, a homodimer of two apparently identical subunits of approximately 180 kDa linked by disulphide bridges. The amino acid sequence of the N-terminus is similar to the Limulus alpha(2)M counterpart. The shrimp alpha(2)M has a wide inhibition spectrum against different proteinase types including trypsin, leucine amino peptidase, chymotrypsin, elastase and papain. The secondary structure of shrimp alpha(2)M is mainly beta-sheet (36%), with a characteristic minimum elipticity at 217 nm. Evidence for a thiolester-mediated inhibition mechanism of proteases by alpha(2)M was provided by inactivation with methylamine. PMID- 12727294 TI - A comparison of hepatoprotective activities of aminoguanidine and N acetylcysteine in rat against the toxic damage induced by azathioprine. AB - Azathioprine (AZA) is an important drug used in the therapy of autoimmune system disorders. It induces hepatotoxicity that restricts its use. The rationale behind this study was the proven efficacy of N-acetylcysteine (NAC; a replenisher of sulfhydryls) and reports on the antioxidant potential of aminoguanidine (AG; an iNOS inhibitor), that might be useful to protect against the toxic implications of AZA. AG (100 mg/kg; i.p.) or NAC (100 mg/kg; i.p.) were administered to the Wistar male rats for 7 days and after that AZA (15 mg/kg, i.p.) was given as a single dose. This caused an increase in the activity of hepatic aminotransferases (AST and ALT) in the serum 24 h after AZA treatment. AZA (7.5 or 15 mg/kg, i.p.) also caused an increase in rat liver lipid peroxides and a lowering of reduced glutathione (GSH) contents. In the other part of experiment, protective effects of AG and NAC were observed on AZA induced hepatotoxicity. NAC significantly protected against the toxic effects produced by AZA. Pretreatment with NAC prevented any change in the activities of both the aminotransferases after AZA. This pretreatment also resulted in a significant decline in the contents of lipid peroxides and a significant elevation in GSH level was evident after AZA treatment. In the group with AG pretreatment the activities of AST and ALT did not increase significantly after AZA when compared to control. However, the lipid peroxides and GSH levels did not have any significant difference when compared to AZA group. These observations also indicate that the improvement in the GSH levels by NAC is the most significant protective mechanism rather than any other mechanistic profile. The protective effect of AG against the enzyme leakage seems to be through the liver cell membrane permeability restoration and is independent of any effects on liver GSH contents. PMID- 12727295 TI - Effects of benzodiazepines on frog ERG. AB - Effect of flurazepam (water-soluble benzodiazepine) on the amplitude and time course of ERG waves was investigated in superfused frog eyecups (Rana ridibunda). Flurazepam (50 and 100 microM) had inhibitory effect on the b- and d-wave amplitude, which was not accompanied with significant changes in their implicit time. Flurazepam potentiated the depressant effect of GABA (2.5 and 5 mM) on the b- and d-wave amplitude. The inhibitory effect of flurazepam was not blocked by 50 microM bicuculline (BCC), (GABA(A) antagonist), although the blocker markedly potentiated the b- and d-wave amplitude. The suppressive effect of flurazepam on the b- but not d-wave amplitude was blocked by 100 microM BCC. Our results indicate existence of functional benzodiazepine regulatory sites on GABA(A) receptors in distal frog retina. PMID- 12727296 TI - Localization of substance P-like immunoreactivity in the palate and trigeminal ganglion of Rana pipiens. AB - This study was undertaken to localize substance P-like immunoreactivity (SP) in the nerve fibers innervating the palate, identify the ganglion of the palatine nerve and determine whether it contains SP cell bodies, in the frog Rana pipiens. The palatine nerve which is a branch of the maxillo-mandibular subdivision of the trigeminal nerve was traced to the trigeminal ganglion that connects to the medulla by the trigeminal nerve root. Using an immunocytochemical method, SP containing fibers with varicosities were found in the connective tissue layer of the palate. Some of these fibers were observed adjacent to blood vessels to the epithelial layer of the palate in apparent innervation of the ciliated epithelial and mucus cells. SP-labeling was also observed in small to medium cells of the trigeminal ganglion. These results appear to support the pharmacological studies of SP on the regulation of mucociliary activity in the frog R. pipiens. PMID- 12727297 TI - Effects of the brominated phenol, lanosol, on cytochrome P-450 and glutathione transferase activities in Haliotis rufescens and Katharina tunicata. AB - The relationship between biochemical transformation mechanisms and dietary preferences has been little studied among marine herbivores. Here we report on basal activities and kinetic parameters of steroid hydroxylase and glutathione transferase from digestive gland tissue of the marine molluscan generalist herbivores Haliotis rufescens and Katharina tunicata and the differential effects of the brominated phenol lanosol [1,2-dihydroxy-3,4-dibromo-5-(hydroxymethyl) benzene] on the activity of these enzymes. Lanosol and other brominated aromatic compounds are prevalent among filamentous red algae frequently consumed by K. tunicata and have been shown to deter feeding in species of Haliotis. Animals were gavaged daily with 10 mg of lanosol per kg of wet mass for 3 days. Mean basal levels of estradiol and testosterone hydroxylase and glutathione transferase specific activities were higher in digestive gland tissue from H. rufescens relative to that of K. tunicata, and only K. tunicata glutathione transferase specific activity was affected by lanosol treatment. Apparent enzyme kinetic parameters (K(m) and V) for the substrate estradiol were higher in K. tunicata, and glutathione transferase from H. rufescens showed a higher efficiency of turnover compared with glutathione transferase from K. tunicata based on V/K(m) ratios. These results suggest a potential relationship between detoxification enzyme induction mechanisms and feeding behaviors among marine herbivores. PMID- 12727298 TI - Effects of proteinase inhibitors on digestive proteinases and growth of the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae). AB - The physiology of the gut lumen of the red flour beetle, T. castaneum, was studied to determine the conditions for optimal protein hydrolysis. Although the pH of gut lumen extracts from T. castaneum was 6.5, maximum hydrolysis of casein by gut proteinases occurred at pH 4.2. The synthetic substrate N-alpha-benzoyl-DL arginine-rho-nitroanilide was hydrolyzed by T. castaneum gut proteinases in both acidic and alkaline buffers, whereas hydrolysis of N-succinyl-ala-ala-pro-phe rho nitroanilide occurred in alkaline buffer. Inhibitors of T. castaneum digestive proteinases were examined to identify potential biopesticides for incorporation in transgenic seed. Cysteine proteinase inhibitors from potato, Job's tears, and sea anemone (equistatin) were effective inhibitors of in vitro casein hydrolysis by T. castaneum proteinases. Other inhibitors of T. castaneum proteinases included leupeptin, L-trans-epoxysuccinylleucylamido [4-guanidino] butane (E-64), tosyl-L-lysine chloromethyl ketone, and antipain. Casein hydrolysis was inhibited weakly by chymostatin, N-tosyl-L-phenylalanine chloromethyl ketone, and soybean trypsin inhibitor (Kunitz). The soybean trypsin inhibitor had no significant effect on growth when it was bioassayed alone, but it was effective when used in combination with potato cysteine proteinase inhibitor. In other bioassays with single inhibitors, larval growth was suppressed by the cysteine proteinase inhibitors from potato, Job's tears, or sea anemone. Levels of inhibition were similar to that observed with E-64, although the moles of proteinaceous inhibitor tested were approximately 1000-fold less. These proteinaceous inhibitors are promising candidates for transgenic seed technology to reduce seed damage by T. castaneum. PMID- 12727299 TI - Structural changes in gills of Lost River suckers exposed to elevated pH and ammonia concentrations. AB - The Lost River sucker (Deltistes luxatus) is a federally listed, endangered fish that occurs primarily in Upper Klamath Lake-a hypereutrophic lake in southern Oregon, USA. A decline of the sucker population in the lake over the past few decades has been partly attributed to adverse water quality conditions, including elevated pH and ammonia concentrations that occur during summer cyanobacterial blooms. We quantitatively analyzed structural changes in gills of larval Lost River suckers after they were exposed to elevated pH and ammonia concentrations for 30 d. Exposure to pH as high as 10 caused no observed structural changes. However, lamellar thickness and O(2) diffusion distance increased significantly (P<0.05) at ammonia concentrations that did not significantly decrease survival, growth, whole-body ion concentrations, or swimming performance. Additionally, we qualitatively observed increases in the frequency of hyperplasic and hypertrophic mucous cells, tissue damage, epithelial lifting, and infiltration of white blood cells into paracellular lymphatic spaces at the highest sublethal ammonia concentration. These observed gill changes typically indicate compromised respiratory and ionoregulatory capacity, although such effects were not manifested in the assays we performed. Regardless, these structural gill changes appear to be a more sensitive indicator of exposure to elevated ammonia concentrations than are more traditional sublethal indices. Therefore, gill histopathology might be a relevant early-warning monitoring tool of the health of Lost River suckers in Upper Klamath Lake, and other species in similar eutrophic systems. PMID- 12727300 TI - Activity of carboxylesterase and glutathione S-transferase in different life stages of carabid beetle (Poecilus cupreus) exposed to toxic metal concentrations. AB - Among the cytoplasmatic enzymes responsible for neutralization of organic xenobiotics, carboxylesterases (CarE) and glutathione S-transferases (GST) play important roles. Our study tested to what extent dietary Zn or Cd could modify the activity of CarE and GST at different life-stages of the carabid beetle Poecilus cupreus. Treatment and stage effects generally were statistically significant. For CarE activity in the beetles exposed to cadmium, only treatment was a significant factor. In all cases, the interaction between studied factors was statistically significant, implying that the physiological condition of the animals may enhance or reduce enzyme activity. We also observed differences between animals treated with cadmium and zinc in the pattern of enzyme activity, and a difference in GST activity measured with two different substrates. Our results confirmed that in studying enzyme activity under metal stress one should consider the animal's life-stage and sex. PMID- 12727301 TI - cDNAs encoding large venom proteins from the parasitoid wasp Pimpla hypochondriaca identified by random sequence analysis. AB - Venom from the parasitoid wasp Pimpla hypochondriaca contains numerous proteins, has potent in vitro anti-haemocytic properties, and disrupts host encapsulation responses. By sequencing 500 cDNAs randomly isolated from a venom gland library, we have identified 60 clones that encode proteins containing potential secretory signal sequences. To identify cDNAs encoding particular venom proteins, N terminal amino acid sequences were determined for large (>30 kDa) venom proteins that had been separated using a combination of gel filtration and SDS-PAGE. We describe five of these cDNAs, which encoded residues that matched with the N terminal sequences of previously undescribed venom proteins. cDNAs vpr1 and vpr3 encoded related proteins of approximately 32 kDa that were found in widely different fractions of gel filtration-separated venom. Neither vpr1 nor vpr3 were closely related to any other protein in the GenBank database, suggesting that they are highly specialised venom components. vpr2 encoded a 57-kDa polypeptide that was similar to a Drosophila protein, of unknown function, which lacks a signal sequence. A fourth clone, tre1, encoded a 61-kDa protein with extensive sequence similarity to trehalases. The 76-kDa sequence encoded by lac1 contained three regions which were very similar to histidine-rich copper-binding motifs, and could be aligned with the laccase from the fungus Coprinus cinereus. This study represents a significant step towards a holistic view of the molecular composition of a parasitoid wasp venom. PMID- 12727302 TI - Effect of glycine in streptozotocin-induced diabetic rats. AB - Inadequate utilization of glucose in diabetes mellitus favors diverse metabolic alterations that play a relevant role in the physio-pathology of chronic complications of this disease. Streptozotocin-induced diabetic rats were treated daily with glycine (130 mM as optimal concentration) or taurine (40 mM) for six months. Groups of diabetic rats without treatment were used as controls. Glucose, total cholesterol, triacylglycerol, and glycated hemoglobin were determined periodically after inducing diabetes. Rats were killed after 6 months of treatment and histological analyses were performed. Diabetic groups that received glycine or taurine showed significant lower concentrations of glucose, total cholesterol, triacylglycerol, and glycated hemoglobin than diabetic control rats (P<0.05) after 6 months treatment. Histological analyses of diabetic rats showed pancreatic atrophy and necrosis, vacuolization, decrease of beta cells, and diffuse glomerulosclerosis. Diabetic rats treated with glycine or taurine showed less enlargement of the glomerular basal membrane than control diabetic rats. Our results suggest that glycine and taurine reduced the alterations induced by hyperglycemia in streptozotocin-induced diabetic rats probably due to inhibition of oxidative processes. PMID- 12727303 TI - The involvement of specific phospholipase C isozymes in catecholamine release from digitonin permeabilized bovine adrenal medullary chromaffin cells. AB - The role of phospholipase C (PLC) in exocytosis has been investigated using digitonin permeabilized, [(3)H]noradrenaline ([(3)H]NA) loaded, bovine adrenal medullary chromaffin cells. The PLC inhibitor U-73122 caused a concentration dependent suppression of Ca(2+)-evoked [(3)H]NA release but increased basal release (that occurring in the absence of Ca(2+)). Preincubation with antibodies against PLCgamma1 or PLCbeta3 (but not PLCdelta1, delta2, beta1 and beta2) also inhibited [(3)H]NA release evoked by Ca(2+) and increased basal release, indicating that only specific PLC isozymes are involved in these actions. Interestingly, PLCgamma1 (but not PLCbeta3) antibodies inhibited the ability of Ca(2+) to increase PLC activity in these permeabilized cells. These data therefore suggest that PLCgamma1 activity may have a specific role in regulating the exocytotic response from the adrenal chromaffin cell. PMID- 12727304 TI - A risk for early-onset Alzheimer's disease associated with the APBB1 gene (FE65) intron 13 polymorphism. AB - Alzheimer's disease (AD) is a genetically complex neurodegenerative disorder and the leading cause of dementia of the elderly. Recently, Hu et al. suggested that a trinucleotide deletion in intron 13 of the APBB1 gene was a factor protecting against late-onset AD. We report here the results of a case/control study aimed at replicating this association. Our study included 461 AD patients and 397 matched controls. We compared the allele and genotype frequencies of the polymorphism between the two groups but did not find any statistically significant difference (P=0.08 and P=0.09, respectively). By contrast, adjusting for age and sex, we found a slight risk associated with the deletion (odds ratio=1.47, 95% confidence interval=1.05-2.04). Stratification by age showed that the risk effect associated with the deletion concerned subjects aged less than 65 years. PMID- 12727305 TI - Static, but not optokinetic visual stimuli induce Fos expression in the retina and brain of retinal degeneration mice. AB - The stimulated expression of the c-fos gene was used in this study to characterize the residual vision of retinal degeneration (rd) mice. Conventional immunohistochemistry was employed to detect Fos-positive nuclei in the rd mouse retina and brain after monocular application of static and dynamic photic stimuli. Diffuse light induced Fos-positive nuclei in the inner nuclear and ganglion cell layers of the stimulated retina and in the contralateral pretectal olivary nucleus. On the other hand, Fos-positive nuclei were rarely detected in the rd mouse retina and brain after stimulation with a slowly displaced random dot pattern. These results suggest that the residual vision of the rd mouse involves mainly the circuits that are related to the processing of static photic stimuli. PMID- 12727306 TI - High expression of nicotinamide N-methyltransferase in patients with idiopathic Parkinson's disease. AB - We have previously speculated that elevated levels of nicotinamide N methyltransferase (NNMT), the primary catabolic enzyme of nicotinamide, may result in reduced Complex I activity in idiopathic Parkinson's disease (IPD) in two ways: (1) reduction in the levels of nicotinamide available for nicotinamide adenine dinucleotide synthesis; and (2) increased methylation of compounds such as tetrahydroisoquinolines and beta-carbolines, which are potent Complex I inhibitors. Expression of NNMT was assessed in 91 cerebella (53 IPD, 38 control) using immunohistochemistry coupled with quantitative digital image analysis. Control cerebella showed a distribution of expression ascribed to low, intermediate and high expressors with ratios of 1:2:1 categories. Expression in the parkinsonian cerebella was significantly higher than in the control group (control group median expression 17%, mean expression 16.6%, range 0-51%, standard deviation 11.4%, standard error 1.9%; IPD group median expression 46%, mean expression 53.7%, range 21-100%, standard deviation 23.4%, standard error 3.2%; P<0.0001; unpaired t-test with Welch correction (parametric) and Mann Whitney U-test (non-parametric)). These results confirm that NNMT expression is elevated in IPD, which may ultimately lead to neurodegeneration via a reduction in Complex I activity. PMID- 12727307 TI - High and low frequency transcutaneous electrical nerve stimulation inhibits nociceptive responses induced by CO2 laser stimulation in humans. AB - The aim of the study was to evaluate the effects of transcutaneous electric nerve stimulation (TENS) on CO(2) laser evoked potentials (LEPs) in 16 normal subjects. The volar side of the forearm was stimulated by 10 Hz TENS in eight subjects and by 100 Hz TENS in the remainder; the skin of the forearm was stimulated by CO(2) laser and the LEPs were recorded in basal conditions and soon after and 15 min after TENS. Both low and high frequency TENS significantly reduced the subjective rating of heat stimuli and the LEPs amplitude, although high frequency TENS appeared more efficacious. TENS seemed to exert a mild inhibition of the perception and processing of pain induced by laser Adelta fibres activation; the implications of these effects in the clinical employment of TENS remain to be clarified. PMID- 12727308 TI - The striopallidal pathway is involved in antiparkinsonian-like effects of the blockade of group I metabotropic glutamate receptors in rats. AB - The aim of the study was to examine the influence of the blockade of group I metabotropic glutamate receptors (mGluRs) on the haloperidol-induced catalepsy and proenkephalin mRNA expression in the rat striatum. Bilateral, intrastriatal injection of AIDA ((RS)-1-aminoindan-1,5-dicarboxylic acid, 3-15 microg/0.5 microl), a selective antagonist of group I mGluRs, inhibited catalepsy induced by haloperidol (0.5 mg/kg i.p.). Repeated intrastriatal AIDA administrations (3 x 15 microg/0.5 microl, 3 h apart) counteracted the haloperidol-induced (3 x 1.5 mg/kg s.c., 3 h apart) increase in the proenkephalin mRNA expression in that structure. The present study indicates that the blockade of the striatal group I mGluRs may inhibit parkinsonian akinesia by normalizing the function of the striopallidal pathway. PMID- 12727309 TI - Additive neuroprotective effect of Ketanserin and Ipsapirone on the hippocampal damage after transient forebrain ischemia in the Mongolian gerbil. AB - Modulation of the serotonin (5HT) system via 5HT1A or 5HT2A receptors exerts a neuroprotective effect on delayed neuronal death after transient forebrain ischemia. We tested the hypothesis that a 5HT1A agonist (Ipsapirone) in combination with a 5HT2A receptor antagonist (Ketanserin) could improve the neuroprotection. Starting 15 min prior to transient forebrain ischemia in the gerbil model, different doses of Ipsapirone (1, 2, 3 mg) and Ketanserin (5 mg/kg) were applied intraperitoneally. Seven days after ischemia, surviving pyramidal cells of the CA1 sector of the hippocampus were counted. The significance of the differences between the means was assessed by an analysis of variance according to the Scheffe test. The hippocampal cell damage was analyzed by histological evaluation. Combined application of Ipsapirone and Ketanserin led to a dose dependent additive effect with up to 83% preservation of hippocampal CA1 neurons (P<0.001). The results of the present study suggest that the combination of 5HT1A receptor agonists and 5HT2A receptor antagonists might be an effective tool for the treatment of cerebral ischemia. PMID- 12727310 TI - Primary cold-sensitive neurons in acutely dissociated cells of rat hypothalamus. AB - Local warming or cooling of the preoptic and anterior hypothalamus (PO/AH) areas evokes various thermoregulatory responses in mammals. We have hypothesized that warm- and cold-sensitive neurons recorded in the PO/AH are multiple thermostats that regulate the core temperature against heat and cold, respectively. However, since the proportion of cold-sensitive neurons is low, it is still controversial whether primary cold-sensitive neurons exist in the PO/AH. To answer this question, we investigated cold-sensitive neurons with Ca(2+) imaging in acutely dissociated PO/AH cells. Their threshold temperatures were 27.3+/-0.44 degrees C (mean+/-SEM, n=55). In extracellular recordings cooling evoked discharges in these cold-sensitive neurons. We conclude that primary cold-sensitive neurons with low threshold temperatures exist in PO/AH. PMID- 12727311 TI - Administration of the 5-hydroxytryptamine(1A) receptor antagonist WAY100635 suppresses acute experimental allergic encephalomyelitis in Lewis rats. AB - Experimental allergic encephalomyelitis (EAE) is a T-cell inflammatory disease of the central nervous system (CNS) widely considered as an animal model of multiple sclerosis. In Lewis rats, myelin basic protein-complete Freund's adjuvant (MBP CFA)-induced EAE is an acute monophasic disease from which animals recover fully. In our experiments, daily treatment (since day 1 after MBP-CFA inoculation) with the 5-hydroxytryptamine((1A)) (5-HT(1A)) receptor agonist (R)-(+)-8-hydroxy-2 (Dipropylamino)-tetralin (R(+)-8-OH-DPAT) resulted in a dose-related enhancement of neurological and histological signs in EAE-induced rats. This effect of R(+)-8 OH-DPAT was reduced by the co-administration of the 5-HT(1A) receptor antagonist (N-[2-(4-[2-mehoxyphenil]-1-piperazinyl)-ethyl]-N-2 pyridinylcyclohexanecarboxamide (WAY100635) at the peak of the acute disease. Moreover, treatment with WAY100635 since inoculation resulted in a delayed onset of the first clinical signs, milder disease and earlier regression of neurological signs along with a decrease in inflammation in the CNS. PMID- 12727312 TI - Phase advancing human circadian rhythms with short wavelength light. AB - The photoreceptor(s) responsible for photoresetting of the human circadian system have not been identified. The aim of the present study was to assess the ability of short wavelength light to alter the timing of circadian rhythms. Eleven male subjects were studied in 15 4-day trials with a single 4 h light pulse administered on day 3, immediately after habitual wake time. The magnitude of the phase shifts in the melatonin acrophase and offset were similar after white (4300 microW/cm(2)) and short wavelength (28 microW/cm(2)) light exposure even though the white light pulse contained 185-fold more photons than the short wavelength light. This finding suggests short wavelength sensitivity of the photoreceptors mediating synchronization of human circadian rhythms. PMID- 12727313 TI - Neuronal and glial inclusions in frontotemporal dementia with or without motor neuron disease are immunopositive for p62. AB - We examined the immunoreactivity of p62 in five cases of frontotemporal dementia (FTD) with ubiquitin-positive, tau-negative inclusions. Only one case had clinical features suggestive of motor neuron disease (MND). In all cases, ubiquitin-positive neuronal inclusions and neurites in the hippocampal region and cerebral neocortex were immunohistochemically positive for p62. Moreover, in the temporal region of a case of FTD with MND, many oligodendrocytes and some astrocytes were positive for p62. These results suggest that the degenerative process involves p62 in FTD and that the process takes place not only in neurons but also in glial cells. PMID- 12727314 TI - Recurrence analysis of human postural sway during the sensory organization test. AB - The present study examined how the availability of and alterations in sensory information during the sensory organization test (SOT) influenced the amount, variability, and temporal structure of spontaneous postural sway in young, healthy adults. Findings indicated that postural sway tended to increase in amount and variability as the SOT condition became increasingly difficult (i.e. as the SOT condition moved from eyes open to eyes closed, to sway-referenced visual surround or support surface, and to sway-reference surface and visual surround). In addition, recurrence quantification analysis revealed that the temporal structure of postural sway tended to become increasingly regular as the SOT condition increased in difficulty. The functional utility of the observed changes in the temporal structure of postural sway across sensory conditions was discussed. PMID- 12727315 TI - Peripheral nerve injury influences the disinhibition induced by focal ischaemia in the rat motor cortex. AB - Photothrombotic lesions were produced in the rat primary motor cortex, and the brain excitability was assessed in a paired-pulse stimulation protocol by transcranial recording, in parallel at 16 points of the frontal cortex, including the insulted and the surrounding areas. The cortical lesion reduced the inhibition in the extended frontal cortex, with a delay of a few minutes. Unilateral facial nerve transection, however, accelerated the widespread disinhibition. Although the mechanism is not clear in detail, both peripheral and central injury-induced disinhibition may have a significant impact on the recovery of the function. PMID- 12727316 TI - A brief and unobtrusive instrument to detect simulation and exaggeration in patients with whiplash syndrome. AB - The objective of this study was to develop and test a brief and unobtrusive instrument to detect exaggeration and simulation in whiplash syndrome. The instrument consists of eight scenarios with ten response options that have to be ordered according to how easy a behavior is to perform. Twenty-five simulating and 25 non-simulating patients with whiplash syndrome of grades 2 and 3 according to the Quebec Task Force classification as well as 25 simulating and 25 non simulating controls completed the instrument. In a cross-validation study 20 controls and 20 patients participated. Malingering and exaggeration scores were determined for each subject and patient. The scores were summed up and compared across malingering and exaggerating subjects and controls and cut-off values were determined to classify the patients. T-tests and a discriminant analysis were used to determine classification accuracy. The instrument correctly identified 94% of the simulators and 84% of the exaggerators in both samples. This brief and unobtrusive instrument can detect exaggeration and simulation in whiplash syndrome. PMID- 12727317 TI - Neuronal nitric oxide synthase (bNOS) messenger RNA and protein expression in developing rat brainstem nuclei. AB - Neuronal nitric oxide synthase (bNOS) messenger RNA expression and immunoreactivity were mapped in series of cryosections through the developing rat brainstem nuclei. Between embryonic day E16 and postnatal day P16, brainstem nuclei expressed both bNOS messenger RNA (mRNA) in situ hybridization signals and protein immunoreactivity. However, NOS mRNA signals were absent from the Edinger Westphal, facial or motory trigeminal nucleus. Strong patterns of mRNA signals and immunoreactivity occurred in neurons located in the substantia nigra pars compacta and the laterodorsal tegmental nuclei. Between E24 and P16, altered patterns of bNOS mRNA positive and immunoreactive neurons, e.g. superior and inferior colliculi, raphe nuclei, solitary tract or pontine nucleus were documented. Altered NOS expression patterns thus may reflect developmental processes within distinct neuronal populations such as cell phenotype discrimination or synaptogenesis within efferent or afferent brainstem pathways. The NOS/NO system therefore appears to be a modulator for intra-/intercellular adjustment processes in normal development. PMID- 12727318 TI - Characterization of the dopamine transporter gene expression and binding sites in cultured human amniotic epithelial cells. AB - In this study we sought to investigate whether the dopamine transporter, DAT, and its binding sites are expressed in the human amniotic epithelial cells (HAEC) using reverse transcription-polymerase chain reaction (RT-PCR) and radioligand binding studies, respectively. The RT-PCR findings showed that HAEC expressed DAT mRNA with 100% homology to the human brain DAT. Saturation binding studies using [3H]mazindol showed a high affinity DAT binding site with K(D) and B(max) values of 12.32+/-1.67 nM and 82.7+/-9.74 fmol/mg protein, respectively. Competition experiments showed that selective DAT blockers are potent displacers of [3H]mazindol binding. The rank order of potency of the competing drugs is consistent with the pharmacology of the DAT. The present results provide compelling evidence that HAEC natively express the DAT mRNA and binding sites. More importantly, these results may suggest that HAEC is an appropriate human cell model for studying dopamine release and uptake processes and potential ligands at these sites. PMID- 12727320 TI - Recruitment gain of antagonistic motoneurons is higher during lengthening contraction than during shortening contraction in man. AB - The purpose of this study was to investigate how the recruitment gain, as derived from the Hoffmann (H)-reflex measurement, of antagonistic motoneurons is modulated during voluntary lengthening (LEN) and shortening (SHO) contractions in the human tibialis anterior (TA) muscle. To this end, the parameters of the ratios of the slope in each ascending part of the H and M recruitment curves (Hslp/Mslp) in the antagonist muscle were compared between LEN and SHO contractions in nine young, healthy subjects. Although there were no differences in the levels of background activity (BGA) between LEN and SHO contractions in the agonist (TA) and antagonist (soleus) muscles, the Hslp/Mslp of the antagonist muscle was significantly higher during LEN contractions than during SHO contractions (P<0.01). It was therefore demonstrated that the recruitment gain of the antagonistic motoneurons to the la afferent inputs was higher during LEN contractions than during SHO contractions despite similar BGA levels. This result might reflect differences in the extent of the reciprocal inhibition from the agonist to the antagonist muscles and/or in the neural mechanism underlying the length-changing manners of the antagonist muscle itself. PMID- 12727319 TI - Stress (hypothalamic-pituitary-adrenal axis) and pain response in male rats exposed lifelong to high vs. low phytoestrogen diets. AB - Estrogens exhibit complex but beneficial effects on brain structure, function and behavior. Soy-derived dietary phytoestrogens protect against hormone-dependent and age-related diseases, due to their estrogen-like hormonal actions. However, the effects of phytoestrogens on brain and behavior are relatively unknown. This study examined the influence of exposing male Long-Evans rats (lifelong) to either a phytoestrogen-rich (Phyto-600) or a phytoestrogen-free (Phyto-free) diet on body weights, behavioral pain thresholds, the hypothalamic-pituitary-adrenal (HPA) hormonal stress response, hippocampal glucocorticoid receptor and brain neural cell adhesion molecules (NCAM) and synaptophysin levels using standard behavioral and biochemical techniques. Body weights were significantly decreased in Phyto-600 fed animals compared to Phyto-free values. There were no significant changes in behavioral pain thresholds, circulating corticosterone concentrations (after acute immobilization stress) or NCAM and synaptophysin levels in various brain regions by the diet treatments. However, Phyto-600 fed males displayed significantly higher plasma adrenocorticotrophin (ACTH) (post-stress) and hippocampal glucocorticoid receptor levels vs. Phyto-free values. These data suggest that (1) body weights are significantly reduced by soy-derived phytoestrogens, (2) behavioral pain thresholds (via heat stimuli) are not influenced by dietary phytoestrogens, but (3) these estrogenic molecules in the hippocampus enhance glucocorticoid receptor abundance and alter the negative feedback of stress hormones towards a female-like pattern of higher ACTH release after activation of the HPA stress axis. This study is the first to show that lifelong consumption of dietary phytoestrogens alters the HPA stress response in male rats. PMID- 12727321 TI - Calcium channel agonist, (+/-)-Bay K8644, causes a transient increase in striatal monoamine oxidase activity in Balb/c mice. AB - We investigated in vivo effects of the L-type calcium channel agonist 1,4-dihydro 2,6-dimethyl-5-nitro-4-[2-(trifluoromethyl) phenyl] pyridine-3-carboxylic acid ((+/-)-Bay K8644) on mitochondrial monoamine oxidase (MAO) activity and striatal dopamine (DA) content employing fluorimetric and HPLC-electrochemical procedures, respectively. (+/-)-Bay K8644 administration resulted in visible behavioral dysfunctions in mice. A reversible dose-independent inhibition of striatal DA levels and a significant increase in the brain mitochondrial MAO-A and -B activities were observed in animals treated with the calcium agonist. A positive relationship between the rise in the enzyme activity and decrease in DA content in (+/-)-Bay K8644 treated animals indicates a direct, but transient effect of this channel activator on DA metabolism, which may be related to acute behavioral syndromes exhibited following administration of the drug. Moreover, a direct involvement of L-type dihydropyridine sensitive calcium channels is indicated in this action, since nicardipine could effectively attenuate (+/-)-Bay K8644 induced behavioral aberrations, or block the striatal DA depletion and the increase in MAO activity. The present results have far-reaching implications in neuropharmacological research, where co-treatment of calcium channel drugs and MAO inhibitors are warranted. PMID- 12727322 TI - GABAergic modulation of noradrenaline release in the median preoptic nucleus area in the rat. AB - Microdialysis was employed to investigate whether gamma-aminobutyric acid (GABA) receptor mechanisms are involved in the regulation of noradrenaline (NA) release in the median preoptic nucleus (MnPO) in awake, freely moving rats. Perfusion with the GABA receptor antagonists as well as agonists was performed in the region of the MnPO through a microdialysis probe and dialysate levels of NA were measured. Perfusion with either bicuculline (10 and 50 microM), a GABA(A) receptor antagonist, or phaclofen (10 and 50 microM), a GABA(B) receptor antagonist, enhanced the release of NA in the MnPO area. Higher-dose perfusion with the GABA(A) agonist muscimol (50 microM) or the GABA(B) agonist baclofen (250 microM) decreased dialysate NA in the MnPO area. An iso-osmotic reduction of fluid volume following subcutaneous treatment with polyethylene glycol (PEG, 30%, 5 ml) significantly increased the NA level in the MnPO area. The increased levels of NA caused by the PEG treatment were attenuated by perfusion with muscimol (10 microM), but not by baclofen (50 microM). These results show the participation of both GABA(A) and GABA(B) receptors in the modulation of the release of NA in the MnPO area, and imply that the GABA(A) receptor mechanism may play an important role in the noradrenergic regulatory system of body fluid balance. PMID- 12727323 TI - Immunocytochemical localization of zinc transporter 3 in the ependyma of the mouse spinal cord. AB - We report, for the first time, the light microscopical and ultrastructural appearance of ZnT3-immunoreactivities in the ependymal cells of the central canal of the mouse spinal cord. Light microscopy revealed the presence of ZnT3 immunoreactive (Ir) ependymal cells in 1 microm thick epon sections stained by the ABC method. The ZnT3-Ir cells were observed at all levels of the spinal cord, but were a little more numerous in lumbosacral segments than in cervicothoracic segments. The ZnT3-Ir cells had large, ovoid nuclei with abundant cytoplasm, and protruded into the lumen of the central canal. Our ultrastructural findings suggest that the ZnT3-Ir ependymal cells possess secretory activity directed towards the central canal. We propose that they may play a role in the trans ependymal mechanism responsible for zinc homeostasis between cerebrospinal fluid and the central area of the gray matter. PMID- 12727324 TI - The mGluR5 selective antagonist 6-methyl-2-(phenylethynyl)-pyridine reduces the spinal neuron pain-related activity in mononeuropathic rats. AB - In rats with chronic constriction of one sciatic nerve (CCI rats), showing behavioural signs of neuropathic pain, 6-methyl-2-(phenylethynyl)-pyridine (MPEP), a selective mGluR5 antagonist, was intraperitoneally administered at 0.75, 1.0 and 1.5 mg/kg or spinally microejected and the effects on the lumbar wide dynamic range neurons activity were investigated. In CCI rats MPEP at 1.0 and 1.5 (but not at 0.75) mg/kg, or spinally microejected induced a significant reduction of the spontaneous (SA) and noxious evoked activity (NEA), and a significant decrease of the suppression of the afterdischarge duration. In sham rats SA was unaffected and NEA was significantly reduced by 1.0 and 1.5 mg/kg MPEP dosages. These findings indicate that the metabotropic GluR5 receptor plays a role in the spinal cord processes underlying neuropathic pain and represents a potential target for new therapeutic approaches. PMID- 12727325 TI - Overexpression of the p75 neurotrophin receptor in the sensori-motor cortex of rats exposed to ethanol during early postnatal life. AB - Foetal alcohol syndrome is a known cause of mental retardation. It has been suggested that the anatomical and functional alterations observed in the cerebral cortex could be mediated by an interference of ethanol with developmental processes modulated by neurotrophins and/or their receptors. We have studied by immunohistochemistry the expression of the p75 neurotrophin receptor (p75 NTR) in the sensori-motor cortex of P10 and P20 rats exposed to the inhalation of ethanol during the first week of postnatal life. At both the studied ages, the number of p75 NTR immunoreactive neurons was higher in ethanol treated animals compared to controls. The increase of immunoreactive elements was relatively more marked in the motor than in the somatosensory cortex. The involvement of p75 NTR in ethanol induced apoptosis and neural plasticity is discussed. PMID- 12727326 TI - Neuroprotective effects of bone marrow stromal cells on rat organotypic hippocampal slice culture model of cerebral ischemia. AB - Organotypic hippocampal slice cultures prepared from newborn rats were maintained in vitro for 9 days. Cultures were then exposed to 30 min of combined oxygen glucose deprivation (OGD). After OGD, the area covered by neurites was decreased. The dead cells of hippocampal slices in the ischemia group were 40.4% at day 3 and 41.6% at day 7 after OGD. The ultrastructure of the CA1 region of the slices was seriously damaged. While hippocampal slices were cultured in combination with bone marrow stromal cells (MSCs), the average area covered by neurites was comparatively increased. The dead cells were only 25.2% at day 3 and 27.1% at day 7 after coculture. The damage of the ultrastructure of the CA1 region in the coculture group was reduced significantly. Thus, in an in vitro model of simulated ischemia, MSCs can promote the outgrowth of neurites from hippocampal slices and alleviate cell damage. The neuroprotective effect might be mediated through diffusible neurotrophic factors secreted from MSCs. PMID- 12727327 TI - Fos protein expression following acute administration of diethyldithiocarbamate in rats. AB - Dithiocarbamates are compounds commonly used in medicine and in agriculture and their prolonged use is known to result in neurotoxicity. Whether this response may be related to early gene expression has not been investigated. We have addressed this issue by mapping Fos expression in rats acutely injected with diethyldithiocarbamate (DDTC) and correlating these data to neural damage in the hippocampus as determined by pyknotic nuclei count. In comparison to saline injected rats, DDTC treatment induced a marked Fos expression in most brain regions at 1 and 3 h. In the hippocampus, a high Fos expression was followed by a variable number of pyknotic nuclei at 6 h, depending on the subregion. The data suggest that, in this model of neurotoxicity, c-fos induction does not reflect a cell commitment to die or survive, but rather a cell response to the DDTC-induced oxidative disorder. PMID- 12727328 TI - Time course of regional brain activations during facial emotion recognition in humans. AB - Recognition of facial expressions of emotions is very important for communication and social cognition. Neuroimaging studies showed that numerous brain regions participate in this complex function. To study spatiotemporal aspects of the neural representation of facial emotion recognition we recorded neuromagnetic activity in 12 healthy individuals by means of a whole head magnetoencephalography system. Source reconstructions revealed that several cortical and subcortical brain regions produced strong neural activity in response to emotional faces at latencies between 100 and 360 ms that were much stronger than those to neutral as well as to blurred faces. Orbitofrontal cortex and amygdala showed affect-related activity at short latencies already within 180 ms after stimulus onset. Some of the emotion-responsive regions were repeatedly activated during the stimulus presentation period pointing to the assumption that these reactivations represent indicators of a distributed interacting circuitry. PMID- 12727329 TI - The antihyperalgesic effect of venlafaxine in diabetic rats does not involve the opioid system. AB - Venlafaxine (VFX) is a structurally novel antidepressant that inhibits reuptake of serotonin and norepinephrine but, unlike tricyclic antidepressants, has few side effects. The present work studies the antihyperalgesic effect of repeated administrations of VFX (five successive injections of 2.5, 5 or 10 mg/kg, s.c., every half-life) in diabetic rats with the paw pressure test and the effect of the opioid receptor antagonist naloxone (1 mg/kg, i.v.) because an opioidergic mechanism is usually considered to be involved in the analgesic effect of antidepressants. VFX induced a significant dose-dependent increase in vocalization thresholds. This effect was not reversed by naloxone. Thus, we demonstrate a clear antinociceptive effect of VFX which, unlike that of most mixed tricyclic antidepressants, does not involve the endogenous opioid system. PMID- 12727330 TI - Increases in spinal vasoactive intestinal polypeptide and neuropeptide Y are not sufficient for the genesis of neuropathic pain in rats. AB - We tested the hypothesis that increases in the spinal levels of vasoactive intestinal polypeptide (VIP) and neuropeptide Y (NPY) were related to the development of neuropathic pain. To this aim, we compared two groups of rats. One group showed well-developed neuropathic pain in the tail following unilateral transection of the inferior and superior caudal trunks between the S1 and S2 spinal nerves, and the other group showed poorly-developed neuropathic pain despite the same nerve injury. The increases in immunoreactivity of VIP and NPY in the S1 dorsal horn (injured segment) were not significantly different between the two groups. These results suggested that increases in the spinal levels of VIP and NPY after peripheral nerve injury were not sufficient for the development of neuropathic pain. PMID- 12727331 TI - The use of formic acid to embellish amyloid plaque detection in Alzheimer's disease tissues misguides key observations. AB - We compared our heat pretreatment method to the widely used formic acid pretreatment technique to immunohistochemically detect amyloid in control and Alzheimer's disease brain tissues. Both methods detected amyloid in plaques, neurons, ependymal cells, circulating monocytes, vascular smooth muscle and endothelial cells. Although there were no observable differences in the intensity of the amyloid labeling in these cell types using both pretreatment methods, there were considerable differences in the intensity of amyloid immunolabeling in the plaques. The formic acid produced much more intense amyloid labeling in the plaques than the heat method. With the heat method, the intensity of the amyloid labeling in the plaques was similar to that detected in nearby neurons suggesting a neuronal origin of plaques. Conversely, the intensity of the amyloid in nearby neurons and plaques was drastically different using the formic acid suggesting unique origins of amyloid. The obvious benefits of formic acid for increasing the sensitivity of amyloid plaque immunolabeling may artifactually emphasize plaques over amyloid-containing cells during analyses. PMID- 12727332 TI - Hypoxia-induced Ca2+-influx in cerebral cortical neuronal nuclei of newborn piglets. AB - The present study was designed to investigate the effect of hypoxia on nuclear calcium-influx in the cerebral cortex of newborn piglets. Anesthetized and ventilated newborn piglets divided into normoxic (n=4) and hypoxic groups with varying degrees of tissue hypoxia (n=10) were studied. Nuclear Ca(2+)-influx was determined using (45)Ca(2+) and plotted against ATP and phosphocreatine levels. The plots were analyzed by non-linear regression (exponential) analysis that showed a curvilinear relationship (r=0.92 for ATP and r=0.88 for phosphocreatine). These data suggest a threshold at which there is a sudden increase in the nuclear calcium-influx that then continues to increase with further decrease in the ATP and phosphocreatine levels. The results demonstrate an increase in the nuclear Ca(2+)-influx during hypoxia in newborn piglets and that this increase correlates in a curvilinear fashion with the increase in the degree of cerebral tissue hypoxia. We propose that the hypoxia-induced increase in intranuclear Ca(2+) is due to altered nuclear membrane Ca(2+)-influx mechanisms and will lead to Ca(2+)-mediated alteration of apoptotic gene expression as well as Ca(2+)-dependent activation of endonucleases that result in DNA fragmentation and subsequent programmed neuronal cell death. PMID- 12727333 TI - Differential cytotoxicity of dopamine and H2O2 in a human neuroblastoma divided cell line transfected with alpha-synuclein and its familial Parkinson's disease linked mutants. AB - alpha-Synuclein accumulates in Lewy bodies and two missense mutations, A30P and A53T, have been linked to familial Parkinson's disease. Neither the normal function of alpha-synuclein nor the pathomechanism of alpha-synuclein-induced neuropathy are known. SK-N-MC neuroblastoma cells were transiently transfected with either wt alpha-synuclein, or its mutants, and their abilities to protect against oxidative stress were assessed. At low expression levels (1 microg cDNA/10(5) cells), all three synuclein variants were devoid of any effect on dopamine-induced cytotoxicity and nitrite production, whereas at higher expression (5 microg cDNA/10(5) cells), the variants enhanced dopamine-mediated effects. Low levels of wt alpha-synuclein blocked H(2)O(2)-induced cytotoxicity and nitrite production, a protective effect that was partly decreased upon higher expression. Both A30P and A53T increased in a dose-dependent manner H(2)O(2) induced nitrite production and cell death. These results show an absence of protective effects for the A30P/A53T mutants, and a differential cytoprotective role of alpha-synuclein against oxidants, which varies according to expression levels. PMID- 12727334 TI - Specific uptake of Abeta1-40 in rat brain occurs in astrocyte, but not in microglia. AB - In the brain of a patient with Alzheimer's disease, beta amyloid peptide (Abeta) is thought to be taken up by glial cells such as astrocyte and microglia to be degraded. However, it is unclear whether the Abeta is absorbed by astrocyte or microglia. The purpose of our study is to determine which type of glial cell, astrocyte or microglia, can take up Abeta. Beta amyloid 1-40 (Abeta1-40) was directly infused into the frontal cortex or hippocampus for 14 days. Dual labeling immunohistochemistry for Abeta1-40 with an astrocytic (GFAP) or microglial (CD11b) marker was performed to examine co-localization of Abeta1-40 and glial markers. In the Abeta1-40 infused site, immunoreactivity of Abeta1-40 was observed only in astrocytes, not in microglia. In addition, Abeta40-1, a reverse peptide of Abeta1-40, was not taken up by astrocytes. These results suggested that the astrocyte-specific uptake of Abeta occurred in the rat brain. PMID- 12727335 TI - Lack of association of interleukin-10 promoter region polymorphisms with Alzheimer's disease. AB - There is increasing evidence that immune mechanisms are involved in the pathogenesis of Alzheimer's disease (AD). Recently, polymorphisms of the interleukin (IL)-1 and IL-6 genes were found to be associated with late-onset AD. The immunoregulatory IL-10 downregulates synthesis of pro-inflammatory cytokines such as IL-1. Current evidence suggests that some polymorphisms in the IL-10 promoter may have functional effects on IL-10 transcription. A total of 406 German AD patients (mean age 70.2+/-10.0 years, range 50-95 years, 42% female) and 251 unrelated non-demented control subjects (mean age 66.8+/-10.6 years, range 50-93 years, 38% female) were investigated for the presence of three polymorphisms in the IL-10 promoter region (-1087A/G, -824C/T, -597C/A). No significant differences in the allelic distribution of the analyzed IL-10 polymorphisms have been found between AD patients and controls. We conclude that polymorphisms in the IL-10 promoter region do not increase the risk of developing AD. PMID- 12727336 TI - Total antioxidant capacity following extrinsic denervation and small intestinal transplantation in the rat. AB - Transplantation of small intestine in a rat model has been shown to affect expression of neurochemicals within enteric inhibitory nerves. However, the mechanism for altered expression of inhibitory neurochemicals is uncertain. Based on our previous studies, we hypothesized that small intestinal transplantation would result in altered intestinal levels of antioxidant capacity. Glutathione, total antioxidant capacity, and lipid peroxide levels were measured at 3 months following (1) transection of rat small intestine, (2) transection and extrinsic denervation of rat intestine, and (3) isotransplantation of rat ileum or (4) allotransplantation of rat ileum with cyclosporine therapy to suppress rejection. Glutathione levels were not significantly different among the four groups. There were trends toward increased lipid peroxide levels following isografting and extrinsic denervation. Total antioxidant capacity was increased following extrinsic denervation (P=0.05). Increased intestinal total antioxidant capacity in response to extrinsic denervation may represent a compensatory mechanism for protection against oxidative stress. This result enhances our understanding of the relationship between tissue antioxidant levels and alteration of enteric nerves. PMID- 12727337 TI - Selective reovirus killing of bladder cancer in a co-culture spheroid model. AB - Up to 50% of the transitional cell carcinomas (TCC) express an activated EGF pathway involving MAP/MEK and RAF kinase thus providing a novel means to selectively eliminate transformed cells expressing such proteins. This EGF pathway expression phenotype was also confirmed in our MGH-U3 and room temperature-112 human TCC cell lines, which makes them a suitable model target for the reovirus oncolysis. We report here on an in vitro assay of co-culture spheroids using either human or rat TCC cells with their corresponding fibroblasts to examine the potential of viral selective lysis for TCC. Reovirus, a respiratory enteric orphan virus, which mammals are exposed to early in life, was used in this study. Selective killing of transformed versus normal cells was assayed by time-lapse photography, vital dye staining, immunohistochemistry, and MTT assay. In this in vitro bladder cancer model, reovirus selectively destroyed the transformed cells by lysis or induction of apoptosis. Based on these findings we have initiated an in vivo pre-clinical study on intravesical administration of reovirus in an animal model to further explore the effect of reovirus-mediated oncolysis of TCC. PMID- 12727338 TI - Analysis of host specificity of two closely related baculoviruses in permissive and nonpermissive cell lines. AB - The baculoviruses Bombyx mori nucleopolyhedrovirus (BmNPV) and Autographa californica multinucleocapsid nucleopolyhedrovirus (AcMNPV) share about 90% identity at the genomic level but they have non-overlapping host range and show a high degree of host specificity. We have demonstrated here that AcMNPV undergoes DNA replication and early gene expression in Bombyx-derived BmN cells but fails to show very late gene expression or produce budded virion (BV) particles. Coinfection with BmNPV supported BV production from AcMNPV in BmN cells at low levels but not very late gene expression or polyhedral inclusion body formation. BV production and very late gene expression from BmNPV, on the contrary, were adversely affected in coinfections. In Spodoptera frugiperda-derived Sf21 cell lines, BmNPV DNA replication, BV production, and very late gene expression took place only when coinfected with AcMNPV. BmNPV exerted a less profound effect on AcMNPV multiplication and very late gene expression in permissive host cell lines. AcMNPV shuts down cellular and viral protein synthesis completely when infected alone or coinfected with BmNPV in BmN cells, whereas BmNPV infection did not affect cellular and viral protein synthesis in Sf21 cells. Overall, AcMNPV showed a more dominant effect by complementing the multiplication of BmNPV in nonpermissive host cells while inhibiting it in BmN cells. PMID- 12727339 TI - Optimization of vesicular stomatitis virus-G pseudotyped feline immunodeficiency virus vector for minimized cytotoxicity with efficient gene transfer. AB - FIV-based lentiviral vector has shown a unique opportunity as an efficient gene delivery vehicle, especially to nondividing human cells. Here, we genetically reconstructed the FIV-based vector by serially deleting residual virus genes of gag and vif, leading to minimized cytotoxicity together with efficient virus production and gene transfer. The modified FIV- based vector was generated by transiently transfecting 293T cells with three plasmids of the gene transfer vector with minimal gag region, the packaging plasmid without vif and the VSV-G expressing plasmid. The vector was routinely generated as many as 1 x 10(7) transducing particles per ml and easily concentrated by simple centrifugation. The cytotoxic effect significantly decreased in sensitive cells to FIV infection even at high multiplicity of infection (MOI), such as 500. Moreover, the transduction efficiency was consistently retained after cell cycle was arrested in a variety of human cells. Taken together, our results suggest that the modified VSV-G pseudotyped FIV-based vector efficiently transduce dividing and nondividing human cells with minimal cytotoxicity. PMID- 12727340 TI - Binding of HTLV-1 virions to T cells occurs by a temperature and calcium dependent process and is blocked by certain type 2 adenosine receptor antagonists. AB - A flow cytometric assay that measures binding of human T-lymphotropic virus type 1 (HTLV-1) virions to target cells was used to investigate the binding process and to screen for compounds affecting viral binding. Results showed that adenosine receptor type 2 antagonists effectively inhibit viral binding at concentrations below 10 microM; no inhibition was seen when antagonist was used to pretreat cells or was added post binding, suggesting direct interference with virus attachment. Efficient HTLV-1 binding required divalent calcium ions and temperatures greater than 20 degrees C. Disruption of lipid rafts by cholesterol depletion compromised viral binding but cleavage of glycosyl-phosphatidylinositol linkages had no effect. A monoclonal antibody (mAb) that recognizes HTLV-1 envelope positions 190-209 impaired binding of virus while other anti-envelope antibodies had no effect. These findings place major constraints on the nature of the HTLV-1 cell binding process and identify a class of inhibitors that may have potential for treatment of infection. PMID- 12727341 TI - Molecular characterization of a dsRNA totivirus infecting the sclerotial parasite Coniothyrium minitans. AB - The complete nucleotide sequence, 4975 bp, of the double-stranded RNA (dsRNA) mycovirus infecting the sclerotial parasite Coniothyrium minitans (CmRV) was determined. Sequence analysis revealed the occurrence of two overlapping open reading frames (ORFs): the 5'-proximal large ORF (ORF1; nucleotide positions 62 2389) encodes a putative coat protein (CP) with a predicted molecular mass of 80344 Da, and the 3'-proximal ORF (ORF2, nucleotide positions 2386-4875) encodes a putative RNA dependent RNA Polymerase (RdRp) with a predicted molecular mass of 82551 Da. The tetranucleotide AUGA at nucleotide positions 2386-2389 includes the predicted start codon of ORF2, which overlaps with the stop codon for ORF1. Based on genome organization and sequence analysis encoded proteins, the virus infecting C. minitans strain Chy-1, designated C. minitans RNA virus (CmRV), belongs to the family Totiviridae. Pairwise sequence comparisons of the deduced amino acid sequences encoded by CmRV as well as phylogentic analysis indicated that it is more closely related to the totiviruses that infect filamentous fungi than to those infecting protozoa, yeast and smut fungi. The role of CmRV in the abnormal phenotype associated with a variant of C. minitans is discussed. PMID- 12727342 TI - Sequence analysis of the N, P, M and F genes of Canadian human metapneumovirus strains. AB - The complete nucleotide sequences of the nucleoprotein (N), phosphoprotein (P), matrix protein (M), and fusion protein (F) genes of 15 Canadian human metapneumovirus (hMPV) isolates were determined. Phylogenetic analysis revealed two distinct genetic clusters, or groups for each gene with additional sequence variability within the individual groups. Comparison of the deduced amino acid sequences for the N, M and F genes of the different isolates revealed that all three genes were well conserved with 94.1-97.6% identity between the two distinct clusters The P gene showed more diversity with 81.6-85.7% amino acid identity for isolates between the two clusters, and 94.6-100% for isolates within the same cluster. PMID- 12727343 TI - Sequence analysis of the medium RNA segment of three Simbu serogroup viruses, Akabane, Aino, and Peaton viruses. AB - The sequence analysis was carried out for the medium (M) RNA segment of the Akabane virus (AKAV), Aino virus (AINV), and Peaton virus (PEAV) of the Simbu serogroup of the genus Orthobunyavirus of the family Bunyaviridae. The complementary sequences of the M RNA segments of AKAV, AINV, and PEAV contain a single large open reading frame (ORF), like other orthobunyaviruses. The ORFs potentially encode 1401 amino acids (aa), 1404 aa, and 1400 aa polypeptides, respectively. The identity of the M segment among these viruses is remarkably low, although previous researchers reported that the small RNA segments are highly conserved. Because the M segment codes for the viral surface glycoproteins G1 and G2, the variability of the M segment may affect the antigenicity of these viruses. Phylogenetic studies based on the M and S segment sequences suggested that genetic reassortment has been occurring among ancestral viruses of the three Simbu serogroup viruses throughout their evolution. PMID- 12727344 TI - Effects of potassium and chloride on ribosome association with the RNA of foot and-mouth disease virus. AB - Foot-and-mouth disease virus (FMDV) and other picornaviruses initiate translation of their polyprotein cap-independently at an internal site of the positive-strand viral RNA. This process is mediated by the internal ribosome entry site (IRES), a highly structured cis-acting RNA element that binds translation initiation factors and ribosomal subunits. During their life cycle, picornaviruses induce proliferation of membrane structures involved in viral replication and an increase in membrane permeability probably facilitating virus progeny release. Here, I analyze the efficiency of association of the ribosomal subunits with the FMDV IRES RNA at elevated salt concentrations. Potassium stimulates FMDV translation, whereas sodium chloride concentrations up to 150 mM neither stimulate nor interfere with FMDV translation. Even high potassium concentrations allow binding of the viral RNA to ribosomes. Chloride stimulates binding of ribosomes to the viral RNA at the stage of 48S initiation complex formation and FMDV translation at concentrations up to 150 mM. Only at elevated concentrations, binding of ribosomal subunits and translation are inhibited by chloride. However, FMDV start site selection is not influenced by potassium salts. These results indicate that the association of the viral RNA with ribosomal subunits is well adapted to high salt conditions that are induced during picornavirus infection. PMID- 12727345 TI - Recognition of cis-acting elements of infectious haematopoietic necrosis virus and viral hemorrhagic septicemia virus by homologous and heterologous helper proteins. AB - Infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicemia virus (VHSV) are grouped in the Novirhabdovirus genus within the family Rhabdoviridae. There are many similarities between these two viruses including the lengths of the leader and trailer regions and the homologies of the terminal sequences. We have developed two systems in which IHNV and VHSV minigenomes encoding the marker green fluorescent protein (GFP) can be expressed from plasmids by T7 RNA polymerase. These negative sense minigenome RNAs can be replicated, transcribed and packaged into infectious particles when coinfected with homologous helper viruses. After infection of the minigenome transfected BHK T7 cells by heterologous helper viruses GFP expression was observed, but packaging of the minigenome RNAs into virus particles did not occur. Packaging of chimeric minigenomes by IHNV and VHSV was also not observed. Cotransfections of the negative sense minigenome plasmids with plasmids encoding nucleoprotein (N), phosphoprotein (P) and RNA polymerase (L) of IHNV and VHSV were carried out in all combinations. Minigenome constructs were expressed only after cotransfection with a set of helper plasmids (N, P and L) all originated from one virus. These results indicate that the cis-acting elements responsible for the encapsidation and transcription were recognized by the homologous and heterologous helper proteins, but packaging of the minigenome RNAs required homologous helper viruses. PMID- 12727346 TI - Deduced amino acid sequence of the small hydrophobic protein of US avian pneumovirus has greater identity with that of human metapneumovirus than those of non-US avian pneumoviruses. AB - We report here the nucleotide and deduced amino acid (aa) sequences of the small hydrophobic (SH) gene of the avian pneumovirus strain Colorado (APV/CO). The SH gene of APV/CO is 628 nucleotides in length from gene-start to gene-end. The longest ORF of the SH gene encoded a protein of 177 aas in length. Comparison of the deduced aa sequence of the SH protein of APV/CO with the corresponding published sequences of other members of genera metapneumovirus showed 28% identity with the newly discovered human metapneumovirus (hMPV), but no discernable identity with the APV subgroup A or B. Collectively, this data supports the hypothesis that: (i) APV/CO is distinct from European APV subgroups and belongs to the novel subgroup APV/C (APV/US); (ii) APV/CO is more closely related to hMPV, a mammalian metapneumovirus, than to either APV subgroup A or B. The SH gene of APV/CO was cloned using a genomic walk strategy which initiated cDNA synthesis from genomic RNA that traversed the genes in the order 3'-M-F-M2 SH-G-5', thus confirming that gene-order of APV/CO conforms in the genus Metapneumovirus. We also provide the sequences of transcription-signals and the M F, F-M2, M2-SH and SH-G intergenic regions of APV/CO. PMID- 12727347 TI - A single amino acid substitution in the cytoplasmic tail of the glycoprotein B of herpes simplex virus 1 affects both syncytium formation and binding to intracellular heparan sulfate. AB - Herpes simplex virus 1 (HSV-1) (S) is a spontaneous syncytial mutant derived from the prototype HSV-1(F) after extensive plaque purification, and produces large syncytial plaques on Vero cells. Marker transfer experiments and DNA sequence analysis mapped the syncytial phenotype to a T-C base substitution at codon 787 of the cytoplasmic domain of mature gB, that results in Leu to Pro substitution and consequently belongs to the syn 3 locus. Both the cytoplasmic and the extracellular domains of gB are active in the fusion event since the addition of anti-gB monoclonal antibodies that recognize the extracellular domain of gB prevent HSV-1(S) induced cell fusion. Similarly, gD also participates in cell fusion since addition of anti-gD monoclonal antibodies also prevent HSV-1(S) induced cell fusion. Furthermore the glycoproteins B and D formed complexes in cells infected with mutant or wild type viruses. The amount of gB bound to total heparan sulfate is lower in the mutant than in the wild type strain. This difference becomes particularly profound when gB is associated with a portion of heparan sulfate intercalated to the membranes. The discrepancy in the binding of the mutant and wild type gB to heparan sulfate may be related to the mechanism of cell fusion induced by HSV-1(S). PMID- 12727348 TI - Mucosally-derived HPV-40 can infect both human genital foreskin and cutaneous hand skin tissues grafted into athymic mice. AB - HPV-40 is a rare HPV type that has been detected only in genital mucosal tissues. This HPV type is very closely related to HPV-7, which has a predominantly cutaneous tissue tropism. We have shown, previously, that an isolate of HPV-40 (described here as HPV-40(Hershey) or HPV-40(H)) productively infected genital tissues. In this study, HPV-40(H) was tested for productive infection of cutaneous tissue. Fetal hand skin fragments were incubated with infectious HPV 40(H) and implanted subrenally into athymic mice. After 120 days, xenografts showed morphological changes consistent with HPV-40(H) infection and were HPV-40 DNA in situ positive and capsid antigen positive. The results demonstrated that hand skin can support HPV-40(H) infection thereby indicating that this viral type has the capacity to infect both genital mucosal and cutaneous tissues. PMID- 12727349 TI - Characterization of a swine-like reassortant H1N2 influenza virus isolated from a wild duck in the United States. AB - An H1N2 influenza virus (A/Duck/North Carolina/91347/01) (Dk/NC) was isolated from a wild duck in the United States in 2001. Genetic analyses showed that this duck virus has the same human/classical swine/avian reassortant genotype as the H1N2 viruses that have been isolated from pigs and turkeys in the US since 1999. Phylogenetic analyses of each gene segment further confirmed that the Dk/NC virus is closely related to the domestic animal H1N2 isolates. In particular, Dk/NC is most closely related to a swine H1N2 virus also isolated in North Carolina. These two viruses and a phylogenetically-defined subset of additional swine H1N2 viruses share a common mutation in the Sb antigenic site on the hemagglutinin protein. The recovery of Dk/NC from a wild bird raises concerns for further widespread distribution of these H1N2 viruses via waterfowl migration. PMID- 12727352 TI - VP7 gene polymorphism of serotype G9 rotavirus strains and its impact on G genotype determination by PCR. AB - Rotaviruses are the single most important etiologic agents of severe diarrhea of infants and young children worldwide. Surveillance of rotavirus serotypes/genotypes (both VP7[G] and VP4[P]) is in progress globally in which polymerase chain reaction (PCR) has been the assay of choice. We investigated polymorphism of the VP7 gene of serotype G9 rotavirus strains and its impact on the determination of VP7 gene genotype by PCR assay. By VP7 gene sequence analysis, we and others have previously shown that the G9 rotavirus strains belong to one of three VP7 gene lineages. By PCR assay using three different sets of commonly used primers specific for G1-4, 8 and 9, 23 Brazilian G9 strains and 5 well-characterized prototype G9 strains which collectively represented all three VP7 gene lineages were typed as: (i) G3; (ii) G4; (iii) G9; (iv) G3 and G9; or (v) G9 and G4 depending on a primer pool employed. This phenomenon appeared to be due to: (i) a VP7 gene lineage-specific polymorphism, more specifically mutation(s) in the primer binding region of the VP7 gene of G9 strain; and (ii) the magnitude of difference in nucleotide homology at respective primer binding site between homotypic (G9) and heterotypic (G3 or G4) primers present in a primer pool employed. PMID- 12727353 TI - Gene organization and homology modeling of two iron superoxide dismutases of the early branching protist Perkinsus marinus. AB - The facultative intracellular oyster parasite, Perkinsus marinus, taxonomically related to both dinoflagellates and apicomplexans, possesses at least two distinct genes (PmSOD1 and PmSOD2) predicted to encode iron-containing superoxide dismutases (FeSOD). DNA blots and sequence analysis suggest that both PmSOD1 and PmSOD2 are single copy and are unlinked. PmSOD1 and PmSOD2 are composed of five and six exons, respectively. All introns are delimited by canonical GT/AG boundaries, and have some features more similar to apicomplexan than dinoflagellate introns. Interestingly, exon 1 of PmSOD2 encodes putative transmembrane and spacer domains with no homology to FeSODs, while exon 2 begins with a methionine codon and is homologous to the N-terminus of FeSODs. The position of introns is not highly conserved between PmSOD1 and PmSOD2, although one intron is in a similar location. Comparison of the intron positions of PmSOD1 and PmSOD2 to those of available apicomplexan FeSODs shows that the intron position shared by PmSOD1 and PmSOD2 is also observed in the FeSOD of Toxoplasma gondii. Comparison of the untranscribed regions 5' and 3' of the coding regions for PmSOD1 and PmSOD2 reveals few motifs in common. Instead, each gene possesses a distinct set of putative upstream transcription factor binding sites. Although the proteins encoded by PmSOD1 and PmSOD2 are only 38% identical to each other, homology modeling indicates that they have nearly identical active site structures. The divergent genomic organizations of two FeSOD genes in the same organism illustrates the complexity of the antioxidant system of even simple, early-branching protists such as P. marinus. PMID- 12727354 TI - Identification and characterization of a novel gene disrupted by a pericentric inversion inv(4)(p13.1q21.1) in a family with cleft lip. AB - Cleft lip with or without cleft palate is a common birth defect affecting 1 in every 700 live births. Several genetic loci are believed to be involved in the pathogenesis of syndromic and non-syndromic clefting. We identified a pericentric inversion of chromosome 4, inv(4)(p13q21) that segregates with cleft lip in a two generation family. By using a combination of fluorescence in situ hybridization, yeast artificial chromosome, bacterial artificial chromosome contig mapping, and database searching we mapped and sequenced the inversion breakpoint region. The pericentric inversion disrupts a gene (ACOD4) on chromosome 4q21 that codes for a novel acyl-CoA desaturase enzyme. The 3.0 kb human ACOD4 cDNA spans approximately 170 kb and is composed of five exons of ACOD4. The inversion breakpoint is located in the second exon. The 3.0 kb mRNA is expressed at high level in fetal brain; a lower expression level was found in fetal kidney. No expression of ACOD4 was detected in fetal lung or liver or in adult tissues. The five exons code for a protein of 330 amino acids, with a predicted molecular weight of 37.5 kDa. The protein is highly similar to acyl-CoA desaturases from Drosophila melanogaster to Homo sapiens. The catalytically essential histidine clusters and the potential transmembrane domains are well conserved. PMID- 12727355 TI - Characterisation of maize peroxidases having differential patterns of mRNA accumulation in relation to lignifying tissues. AB - Among other enzymes, peroxidases have been proposed to participate in the latest steps of lignin biosynthesis. In order to identify new proteins involved in such mechanism of lignification in maize, we have isolated three cDNAs coding for three different peroxidases, named ZmPox1, ZmPox2, and ZmPox3, respectively. Computational analyses of these three proteins correlate with features typically attributed to heme-containing plant peroxidases of approximately 300 amino acid residues. Although with different expression levels, ZmPox2 and ZmPox3 mRNAs are accumulated in the elongating region of young roots but not in the root tips. In addition, the ZmPox2 mRNA levels are up-regulated by wounding and ethylene treatments. However, ZmPox1 is also expressed in the root tip meristems, where lignification does not occur. Finally, in situ hybridisations indicate that ZmPox2 mRNA localises in vascular tissues and epidermis. Although ZmPox1 mRNA localises in the same regions as ZmPox2 mRNA in root tips, its mRNA is only detected in the epidermis but not in the vascular tissues of young roots, suggesting that the function of ZmPox1 is not correlated to lignification. In addition, although ZmPox3 mRNA is also detected in the regions where lignification occurs, the involvement of this peroxidase in such a mechanism remains to be further investigated due to its very low expression level. Therefore, based on its amino acid sequence and mRNA accumulation and localisation patterns, the involvement of ZmPox2 in the latest steps of lignification is discussed. PMID- 12727356 TI - Distinctiveness of the genomic sequence of Shiga toxin 2-converting phage isolated from Escherichia coli O157:H7 Okayama strain as compared to other Shiga toxin 2-converting phages. AB - Shiga toxin 2-converting phage was isolated from Escherichia coli O157:H7 associated with an outbreak that occurred in Okayama, Japan in 1996 (M. Watarai, T. Sato, M. Kobayashi, T. Shimizu, S. Yamasaki, T. Tobe, C. Sasakawa and Y. Takeda, Infect. Immun. 61 (1998) 3210-3204). In this study, we analyzed the complete nucleotide sequence of Shiga toxin 2-converting phage, designated Stx2phi-I, and compared it with three recently reported Stx2-phage genomes. Stx2phi-I consisted of 61,765 bp, which included 166 open reading frames. When compared to 933W, VT2-Sakai and VT2-Sa phages, six characteristic regions (regions I-VI) were found in the Stx2 phage genomes although overall homology was more than 95% between these phages. Stx2phi-I exhibited remarkable differences in these regions as compared with VT-2 Sakai and VT2-Sa genes but not with 933W phage. Characteristic repeat sequences were found in regions I-IV where the genes responsible for the construction of head and tail are located. Regions V and VI, which are the most distinct portion in the entire phage genome were located in the upstream and downstream regions of the Stx2 operons that are responsible for the immunity and replication, and host lysis. These data indicated that Stx2phi-I is less homologous to VT2-Sakai and VT2-Sa phages, despite these three phages being found in the strains isolated at the almost same time in the same geographic region but closely related to 933W phage which was found in the E. coli O157 strain 933W isolated 14 years ago in a different geographic area. PMID- 12727357 TI - Hypomutable regions of yeast TFIIB in a unigenic evolution test represent structural domains. AB - As genome sequences of many organisms - including humans - are being decoded, there is a great need for genetic tools to analyze newly discovered genes/proteins. A 'unigenic evolution' approach has been previously proposed for dissecting protein domains, which is based on the assumption that functionally important regions of a protein may tolerate missense mutations less well than other regions. We describe a unigenic evolution analysis of general transcription factor IIB (TFIIB) - a protein that is well characterized both structurally and functionally - to better understand the molecular basis of this genetic approach. The overall distribution profile of hypomutable regions within yeast TFIIB correlates extremely well with the known compact structural domains, suggesting that the unigenic evolution approach can help reveal structural properties of a protein. We further show that a small region located immediately carboxyl terminal to the zinc ribbon motif is functionally important despite its strong hypermutability. Our study further demonstrates the usefulness of the unigenic evolution approach in dissecting protein domains, but suggests that the mutability of different regions of a protein in such a test is determined primarily by their structural properties. PMID- 12727358 TI - Molecular cloning and expression analysis of a novel caspase recruitment domain protein (CARD) in common carp Cyprinus carpio L. AB - A novel caspase recruitment domain protein (CARD) was isolated from common carp Cyprinus carpio L. by expressed sequence tag analysis. This gene consist of a 2016 bp open reading frame and untranslated regions, which is putatively translated to a protein of 535 amino acid residues. The gene harbors domains (CARD and Coiled-coil domain), which are conserved in proteins of CARD family. The CARD domain have carp was similar to human CARD9 with 72.4% identity. Expression analysis revealed that CARD gene of carp (carp-CARD) expressed in normal tissues of head kidney, spleen, liver, heart and brain. Here we demonstrated that the expression of carp-CARD increased by cortisol treatment in all the tissues and had a high and long lasting expression in cortisol treated spleen. PMID- 12727359 TI - Endotoxic and immunobiological activities of a chemically synthesized lipid A of Helicobacter pylori strain 206-1. AB - A synthetic lipid A of Helicobacter pylori strain 206-1 (compound HP206-1), which is similar to its natural lipid A, exhibited no or very low endotoxic activities as compared to Escherichia coli-type synthetic lipid A (compound 506). Furthermore, compound HP206-1 as well as its natural lipid A demonstrated no or very low mitogenic responses in murine spleen cell. On the other hand, compound HP206-1 showed a weaker but significant production of interleukin-8 in a gastric cancer cell line, MKN-1, in comparison with compound 506. Furthermore, compound HP206-1 exhibited induction of tumor necrosis factor-alpha production in human peripheral blood mononuclear cells and the cytokine production was clearly inhibited by mouse anti-human Toll-like receptor (TLR) 4 monoclonal antibody HTA125. Our findings indicate that the chemically synthesized lipid A, mimicking the natural lipid A portion of lipopolysaccharide from H. pylori strain 206-1, has a low endotoxic potency and immunobiological activities, and is recognized by TLR4. PMID- 12727360 TI - Antibacterial actions of fatty acids and monoglycerides against Helicobacter pylori. AB - The bactericidal potencies of saturated and unsaturated fatty acids (FAs) and monoglycerides (MGs) against Helicobacter pylori were determined following short incubations with freshly harvested cells over a range of pHs. FAs and their derivatives with an equivalent-carbon number of 12 were the most potent: lauric acid had a minimum bactericidal concentration (MBC) at pH 7.4 of 1 mM, myristoleic and linolenic acid were the most potent unsaturated FAs (MBCs of 0.5 mM, pH 7.4), and monolaurin was the most potent MG (MBC 0.5 mM). Potencies of saturated FAs were increased sharply by lowering pH, and a decrease of only 0.5 pH units can cause a change from non-lethal to lethal conditions. Conversely, the bactericidal action of monolaurin was not pH-dependent. The bactericidal potencies of unsaturated FAs increased with degree of unsaturation. When more than one FA or FA plus MGs were present, their combined action was additive. Urea and endogenous urease did not protect H. pylori from the bactericidal action of FAs. These results suggest that H. pylori present in the stomach contents (but not necessarily within the mucus barrier) should be rapidly killed by the millimolar concentrations of FAs and MGs that are produced by pre-intestinal lipase(s) acting on suitable triglycerides such as milk fat. PMID- 12727361 TI - Macrophage colony-stimulating factor enhances phagocytosis and oxidative burst of mononuclear phagocytes against Penicillium marneffei conidia. AB - The responses of rabbit pulmonary alveolar macrophages (PAMs) and elutriated human monocytes (EHMs) to Penicillium marneffei, an emerging dimorphic fungus that may cause fatal disease in human immunodeficiency virus-infected patients, were studied. PAMs and EHMs comparably phagocytosed conidia of two P. marneffei strains in the presence of serum. Electron microscopy showed intraphagosomal destruction of conidia after 12 h. Serum-opsonized conidia elicited significantly more superoxide anion (O(2)(-)) release from EHMs compared to non-opsonized conidia, but equivalent O(2)(-) amounts to that elicited by serum-opsonized Aspergillus fumigatus conidia. Macrophage colony-stimulating factor (M-CSF) significantly enhanced phagocytosis of P. marneffei conidia by PAMs and EHMs, as shown by light microscopy. Moreover, M-CSF enhanced O(2)(-) production by EHMs in response to both serum-opsonized (P<0.001) and non-opsonized (P=0.03) conidia of A. fumigatus as well as conidia of the P. marneffei isolates (P<0.001 and 0.03). We conclude that M-CSF enhances phagocytosis and oxidative metabolism of mononuclear phagocytes suggesting a potential role for this cytokine in host defense against pulmonary and disseminated P. marneffei infection. PMID- 12727362 TI - Detection of gene mutations related with drug resistance in Mycobacterium leprae from leprosy patients using Touch-Down (TD) PCR. AB - The lack of methods to identify Mycobacterium leprae with the resistance against multi-drugs quickly and specifically has hindered effective chemotherapy against M. leprae infection. To screen M. leprae with resistance against multi-drugs, the Touch-Down (TD)-PCR has been used in this study. Sequences of the folP, rpoA, B, and gyrA, B genes were analyzed for isolates of M. leprae from leprosy patients in Korea. We amplified designated region of several genes in M. leprae involved in drug resistance and could obtain the PCR products of each gene. The mutations in the particular region of folP, rpoB, and gyrB gene were certified by TD-PCR single-stranded conformational polymorphism and DNA sequencing, respectively. PMID- 12727363 TI - Differential microorganism-induced mannose-binding lectin activation. AB - Mannose-binding lectin (MBL) is a serum complement factor playing a dominant role in first-line defense. When MBL binds to specific sugar moieties on microorganisms, the lectin complement pathway (LCP) is activated. Changes in the mbl gene and promotor may result in MBL with less activity, predisposing the individual to recurrent infections. Using a functional MBL assay, we investigated at what concentration different microbes activated MBL. Less than 1 colony forming unit (CFU) of Neisseria meningitidis groups B and C still activated MBL, which may be ascribed to filterable blebs. Nocardia farcinica and Legionella pneumophila activated MBL well, which raises new questions about host susceptibility. In contrast to other research, Pseudomonas aeruginosa activated the LCP potently. PMID- 12727364 TI - Incidence of haemolysin-positive and drug-resistant Aeromonas hydrophila in freshly caught finfish and prawn collected from major commercial fishes of coastal South India. AB - The incidence of Aeromonas hydrophila in freshly caught finfish and prawns from four major commercial fish landing sites of coastal South India was studied for a period of one year. Among 514 analysed samples of seafood (410 finfish and 104 prawn), 37% of them (37.3% of finfish and 35.6% of prawn) were contaminated with A. hydrophila. A total of 255 strains of A. hydrophila were isolated. Of the total isolates, about 78.4% of them were producers of haemolysin. All strains were resistant to bacitracin and all were sensitive to chloramphenicol. The results indicate that the strains originated from high-risk sources. The presence of A. hydrophila is an indication of marine contamination. The increasing presence of haemolysin-producing multiple drug-resistant A. hydrophila in fish and prawn may become a potential human health hazard. PMID- 12727365 TI - 6,6'-Dimycoloyl trehalose from a rapidly growing Mycobacterium: an alternative antigen for tuberculosis serodiagnosis. AB - Mycobacterial O-acyltrehaloses have been described as highly specific and sensitive reagents for tuberculosis immunodiagnosis. An O-acyltrehalose containing lipid fraction from the rapidly growing Mycobacterium fortuitum was found to include additional antigens, which presented high cross-reactivity with sera from tuberculosis-infected patients. Based on a combination of selective chemical degradations, thin-layer-chromatography analyses and (1)H-nuclear magnetic resonance spectroscopy, the antigenic by-product was identified as 6,6' dimycoloyl trehalose, the so-called cord factor. The lipid was purified and tested in ELISA for pulmonary tuberculosis serodiagnosis. Sensitivity and specificity of the test were found to be 66.6-74.1% and 95.2-99.0%, respectively, showing a slightly higher efficiency as compared to the ELISA performed using 6,6'-dimycoloyl trehalose from Mycobacterium tuberculosis. No cross-reactivity was found with sera from Nocardia-infected individuals. PMID- 12727366 TI - HUVEC take up opsonized zymosan particles and secrete cytokines IL-6 and IL-8 in vitro. AB - Uptake of zymosan A particles by human umbilical vein endothelial cells (HUVEC) and its effect on cellular cytokine and oxygen radical production was examined. HUVEC took up more serum-opsonized than -unopsonized zymosan as demonstrated by flow cytometry with fluorescence-labeled particles. The former uptake was inhibited in the presence of anti-C3c antibodies and thus complement-mediated. It probably occurred via CR1 (CD35), although participation of other receptors cannot be ruled out. Scanning electron microscopy indicated that HUVEC with fully internalized zymosan particles were damaged. Prolonged incubation of both serum opsonized and -unopsonized zymosan particles with HUVEC induced increased secretion of the proinflammatory cytokines IL-6 and IL-8 to the cell culture supernatants, but had no effect on production of oxygen radicals. The results confirm previous reports that EC can internalize yeast and other pathogens and points to complement as a mechanism of uptake, but illustrates that the cells may be damaged in the process. Moreover, EC may participate in the anti-infection defense effort by secreting proinflammatory and chemotactic cytokines in response to the contact with pathogens. PMID- 12727367 TI - The CD14 receptor does not mediate entry of Mycobacterium tuberculosis into human mononuclear phagocytes. AB - Prior reports have suggested that CD14 mediates uptake of Mycobacterium tuberculosis into porcine alveolar macrophages and human fetal microglia, but the contribution of CD14 to cell entry in human macrophages has not been studied. To address this question, we used flow cytometry to quantify uptake by human monocytes and alveolar macrophages of M. tuberculosis expressing green fluorescent protein. Neutralizing anti-CD14 antibodies did not affect bacillary uptake and the efficiency of bacillary entry was similar in THP-1 cells expressing low and high levels of CD14. However, most internalized bacteria were found in CD14+ but not in CD14- monocytes because M. tuberculosis infection upregulated CD14 expression. We conclude that: (1) CD14 does not mediate cellular entry by M. tuberculosis; (2) M. tuberculosis infection upregulates CD14 expression on mononuclear phagocytes, and this may facilitate the pathogen's capacity to modulate the immune response. PMID- 12727368 TI - The identity of the O-specific polysaccharide structure of Citrobacter strains from serogroups O2, O20 and O25 and immunochemical characterisation of C. youngae PCM 1507 (O2a,1b) and related strains. AB - Serological studies using SDS-PAGE and immunoblotting revealed that from five strains that are ascribed to Citrobacter serogroup O2, four strains, PCM 1494, PCM 1495, PCM 1496 and PCM 1507, are reactive with specific anti-Citrobacter O2 serum. In contrast, strain PCM 1573 did not react with anti-Citrobacter O2 serum and, hence, does not belong to serogroup O2. The LPS of Citrobacter youngae O2a,1b (strain PCM 1507) was degraded under mild acidic conditions and the O specific polysaccharide (OPS) released was isolated by gel chromatography. Sugar and methylation analyses along with (1)H- and (13)C-NMR spectroscopy, including two-dimensional (1)H,(1)H COSY, TOCSY, NOESY and (1)H,(13)C HSQC experiments, showed that the repeating unit of the OPS has the following structure: [structure: see text]. NMR spectroscopic studies demonstrated that Citrobacter werkmanii O20 and C. youngae O25 have the same OPS structure as C. youngae O2. Sugar and methylation analyses of the core oligosaccharide fractions demonstrated structural differences in the lipopolysaccharide core regions of these strains, which may substantiate their classification in different serogroups. PMID- 12727369 TI - C3/Ig and Ig/C3 two-component-determined circulating immune complexes (TCIC) in patients with HCV infection. AB - In the present study, we measured the levels of immunoglobulin (Ig)- and complement 3 (C3)-determined circulating immune complexes (two-component determined CIC, or TCIC) in hepatitis C virus (HCV)-infected patients. TCIC was dissected into C3/Ig-TCIC and Ig/C3-TCIC by a reciprocal use of coating and detecting antibodies. The current study was carried out in 117 infected HCV patients and 252 healthy controls. We found that C3/Ig-TCIC elevation was a common feature in patients with HCV infection. Positive rates and levels of C3/IgG-TCIC and C3/IgM-TCIC were significantly higher in the patients with abnormal alanine aminotransferase (ALT) than patients with normal ALT (70.6% vs. 17.0%, 0.56 OD vs. 0.47 OD and 0.71 OD vs. 0.65 OD, respectively, P<0.001). However, the levels of IgM/C3-TCIC and IgA/C3-TCIC were significantly higher in individuals with HCV infection than in healthy controls, whereas the level of IgG/C3-TCIC was significantly lower in the former group than in the latter group. In summary, our results suggest that IgG and C3 TCIC may play an important role in liver cell injury during the course of HCV infection and may be a hallmark for hepatitis C pathogenesis. Elevated C3/Ig-TCIC, accompanied by decreased Ig/C3 TCIC, forms a peculiar trait in HCV infection. Our findings thus provide new insights into HCV pathogenesis. PMID- 12727370 TI - Characterisation of the immune response to the UK human anthrax vaccine. AB - The UK human anthrax vaccine consists of the alum-precipitated culture supernatant of Bacillus anthracis Sterne. In addition to protective antigen (PA), the key immunogen, the vaccine also contains a number of other bacteria- and media-derived proteins. These proteins may contribute to the transient side effects experienced by some individuals and could influence the development of the PA-specific immune response. Bacterial cell-wall components have been shown to be potent immunomodulators. B. anthracis expresses two S-layer proteins, EA1 and Sap, which have been demonstrated to be immunogenic in animal studies. These are also immunogenic in man so that convalescent and post-immunisation sera contain specific antibodies to Ea1, and to a lesser extent, to Sap. To determine if these proteins are capable of modifying the protective immune response to PA, A/J mice were immunised with equivalent amounts of recombinant PA and S-layer proteins in the presence of alhydrogel. IgG isotype profiles were determined and the animals were subsequently challenged with spores of B. anthracis STI. The results suggest that there was no significant shift in IgG isotype profile and that the presence of the S-layer proteins did not adversely affect the protective immune response induced by PA. PMID- 12727371 TI - The evolving epidemiology of invasive meningococcal disease: a two-year prospective, population-based study in children in the area of Athens. AB - In response to an increase in the incidence in invasive meningococcal disease (IMD) due to Neisseria meningitidis, a system of hospital- and laboratory-based surveillance was used in a prospective epidemiological and clinical assessment of IMD in children 0-13 years of age hospitalized in the Athens area between 1 January 1999 and 31 December 2000. The annual incidence of laboratory-confirmed disease was 10.2/100,000. Serogroup B strains were predominant. There was a sharp decrease in serogroup C from 19% of cases in 1999 to 3% in 2000 (P=0.013). Of note was the emergence of serogroup A responsible for 7% of the cases. The overall case fatality rate was 4.5%, but 2.8% for microbiologically confirmed cases. A remarkable decrease in disease severity assessed by admissions to intensive care units was noted during the second study year. Polymerase chain reaction-based methods for detection of meningococcal DNA were the sole positive laboratory test in 45% of the cases and the only test on which serogroup determination was based in 52% of groupable cases. The epidemiological and clinical profile of meningococcal disease appears to be rapidly evolving and close monitoring is required particularly for input into decisions regarding use of meningococcal vaccines. PMID- 12727372 TI - Relationship between haemolytic and sphingomyelinase activities in a partially purified beta-like toxin from Staphylococcus schleiferi. AB - beta-Toxins of staphylococcal species possess dual activity in that they can both lyse erythrocytes (by 'hot-cold' lysis) and catalyse hydrolysis of membrane associated sphingomyelin. However, the precise relationship between these two activities has not been extensively studied. We have partially purified a beta like toxin from culture supernatants of Staphylococcus schleiferi N860375 which exhibits both 'hot-cold' lysis of erythrocytes and neutral sphingomyelinase activities. This toxin has a strong preference for sheep erythrocytes, the membranes of which are rich in sphingomyelin. Kinetic analysis suggests that haemolysis and sphingomyelinase activities are very closely associated obeying identical Michaelis-Menten kinetics. However, pre-treatment with antibodies to Staphylococcus aureus beta-toxin, Ca(2+), dithiothreitol and phenylmethylsulfonyl fluoride appear to inhibit sphingomyelinase activity significantly more strongly than haemolysis while Mg(2+) activates sphingomyelinase activity more strongly than haemolysis. We attribute these effects to differences in binding properties in the two assays. Micropurification by both sphingosylphosphocholine-agarose affinity chromatography and preparative electrophoresis revealed that the 34-kDa toxin associates non-covalently with individual proteins. PMID- 12727374 TI - Genetic analysis of Streptococcus agalactiae strains isolated from neonates and their mothers. AB - Streptococcus agalactiae or group B streptococcus (GBS) is the most common cause of neonatal sepsis and meningitis in neonates. One of the major questions is whether the GBS strains able to cause neonatal invasive disease have peculiar genetic features. A collection of S. agalactiae strains, isolated from cervix, vagina and rectum of 10 mothers and from throat, ear and umbilicus of their newborns was genetically characterized by pulsed-field gel electrophoresis (PFGE). This study demonstrated that the strains isolated from each mother and her child were all genetically identical but that the strains from the 10 mother/child pairs mutually were genetically heterogeneous and 10 different PFGE patterns were found. Although it has been suggested that PFGE would be able to identify virulence traits to direct decisions in antibiotic management, the heterogeneous feature of GBS strains does not support broad application. PMID- 12727373 TI - Evaluation of Proteus mirabilis structural fimbrial proteins as antigens against urinary tract infections. AB - Proteus mirabilis is a common cause of urinary tract infection (UTI) and produce several types of different fimbriae, including mannose-resistant/Proteus-like fimbriae, uroepithelial cell adhesin (UCA), and P. mirabilis fimbriae (PMF). Different authors have related these fimbriae with different aspects of P. mirabilis pathogenesis, although the precise role of fimbriae in UTI has not yet been elucidated. In this work we expressed and purified recombinant structural fimbrial proteins of these fimbriae (MrpA, UcaA, and PmfA) and assessed their role as protective antigens using an ascending and a haematogenous model of UTI in the mouse. MrpA protected subcutaneously immunised mice in both models, suggesting that it could be taken into account as a promising vaccine candidate against P. mirabilis UTI. UcaA could also be an interesting subunit to be studied although it only protected mice that were challenged intravenously. All subunits elicited a strong specific serum IgG response but there was no significant correlation between antibody levels and protection. Only PmfA-immunised mice elicited a significant urinary antibody response but this protein was unable to confer protection against P. mirabilis experimental challenges. These results may contribute to the development of vaccines against P. mirabilis, an important cause of complicated UTI. PMID- 12727375 TI - Silencing unhealthy alleles naturally. AB - Recently, it has been reported that small interfering RNAs can silence mutant alleles of genes while not affecting wild-type alleles. This high specificity should also limit off-target effects when these molecules are used in drug-target validation or as therapeutics. Small RNA technology is both extremely versatile, because virtually any expressed gene can be inhibited, and effective, because it exploits natural cellular machinery that evolved to use small RNAs to regulate gene expression. PMID- 12727376 TI - 'Super bugs' for bioremediation. AB - Chlorinated organic compounds are among the most significant pollutants in the world. Sequential use of anaerobic halorespiring bacteria, which are the key players in biological dehalogenation processes, and aerobic bacteria whose oxygenases are modified by directed evolution could lead to efficient and total degradation of highly chlorinated organic pollutants. Recently three interesting papers on halorespiration and polychlorinated biphenyl biodegradation were published. PMID- 12727377 TI - Transgenesis and yield: what are our targets? AB - Plant metabolic engineering has been used to generate a wide range of transgenic lines in which specific metabolic steps have been targeted. Attempts to increase yield of some agronomically important crops using this approach have highlighted the inherent complexities of modulating plant metabolism. In light of the recent findings by Regierer et al. this article addresses the major challenges faced with respect to enhancing yield through transgenesis. PMID- 12727378 TI - Weeding with transgenes. AB - Transgenes promise to reduce insecticide and fungicide use but relatively little has been done to significantly reduce herbicide use through genetic engineering. Recently, three strategies for transgene utilization have been developed that have the potential to change this. These are the improvement of weed-specific biocontrol agents, enhancement of crop competition or allelopathic traits, and production of cover crops that will self-destruct near the time of planting. Failsafe risk mitigation technologies are needed for most of these strategies. PMID- 12727380 TI - Isolating ligands specific for human vasculature using in vivo phage selection. AB - The endothelium lining blood vessels expresses molecules that are restricted in their expression to a particular tissue or organ. These molecules are attractive targets for therapy and diagnosis because they allow agents to be delivered specifically to the blood vessels supplying the desired tissue. However, it is difficult to identify these tissue-specific molecules because endothelium loses much of its tissue-specific nature when it is removed from the organ. This can be overcome by using in vivo phage selection - injecting libraries of phage bearing antibodies or peptides into an animal and isolating phage that bind to the relevant tissue. A variation on this approach, in which in vivo phage selection is performed in animals bearing human tissue xenografts, allows the isolation of peptides (and presumably other molecules) specific for human vasculature. PMID- 12727379 TI - Development of network-based pathway definitions: the need to analyze real metabolic networks. PMID- 12727381 TI - Biochemical engineering: cues from cells. AB - Engineering principles are used in the exploitation of biocatalysts derived from cells. The purity of reagents, catalysts and maintenance of operation variables are extremely important for bioengineering systems. Any change in the purity of reagents or in operation variables usually leads to a dramatic decrease in productivity. Cellular systems, however, are able to work with relatively high impure conditions and increase their productivity in response to external signals. Thus the seemingly disordered 'bag of juice' or cytoplasm has more order and much higher order of integration than first appears. Learning the semantics of this paradoxical ability of order and integration would help bioengineers to understand and enhance productivity even using impure reagents. PMID- 12727382 TI - Haematococcus astaxanthin: applications for human health and nutrition. AB - The carotenoid pigment astaxanthin has important applications in the nutraceutical, cosmetics, food and feed industries. Haematococcus pluvialis is the richest source of natural astaxanthin and is now cultivated at industrial scale. Astaxanthin is a strong coloring agent and a potent antioxidant - its strong antioxidant activity points to its potential to target several health conditions. This article covers the antioxidant, UV-light protection, anti inflammatory and other properties of astaxanthin and its possible role in many human health problems. The research reviewed supports the assumption that protecting body tissues from oxidative damage with daily ingestion of natural astaxanthin might be a practical and beneficial strategy in health management. PMID- 12727383 TI - Bacterial replacement therapy: adapting 'germ warfare' to infection prevention. AB - The individual bacterial members of our indigeneous microbiota are actively engaged in an on-going battle to prevent colonisation and overgrowth of their terrain by competing microbes, some of which might have pathogenic potential for the host. Humans have long attempted to intervene in these bacterial interactions. Ingestion of probiotic bacteria, particularly lactobacilli, is commonly practiced to promote well-balanced intestinal microflora. As bacterial resistance to antimicrobials has increased, so too has research into colonisation of human tissues with specific effector strains capable of out-competing known bacterial pathogens. Recent progress is particularly evident in the application of avirulent Streptococcus mutans to the control of dental caries, alpha hemolytic streptococci to reduction of otitis media recurrences and Streptococcus salivarius to streptococcal pharyngitis prevention. PMID- 12727384 TI - Manipulation of the mouse genome: a multiple impact resource for drug discovery and development. AB - Few would deny that the pharmaceutical industry's investment in genomics throughout the 1990s has yet to deliver in terms of drugs on the market. The reasons are complex and beyond the scope of this review. The unique ability to manipulate the mouse genome, however, has already had a positive impact on all stages of the drug discovery process and, increasingly, on the drug development process too. We give an overview of some recent applications of so-called 'transgenic' mouse technology in pharmaceutical research and development. We show how genetic manipulation in the mouse can be employed at multiple points in the drug discovery and development process, providing new solutions to old problems. PMID- 12727385 TI - Hematopoietic stem cells: from the bone to the bioreactor. AB - The ex vivo expansion of human hematopoietic stem cells is a rapidly developing area with a broad range of biomedical applications. The mechanisms of renewal, differentiation and plasticity of stem cells are currently under intense investigation. However, the complexity of hematopoiesis, the heterogeneity of the culture population and the complex interplay between the culture parameters that significantly influence the proliferation and differentiation of hematopoietic cells have impaired the translation of small scale results to the highly demanded large-scale applications. The better understanding of these mechanisms is providing the basis for more rational approaches to the ex vivo expansion of hematopoietic stem cells. Efforts are now being made to establish a rational design of bioreactor systems, allowing the modeling and control of large-scale production of stem cells and the study of their proliferation and differentiation, under conditions as similar as possible to those in vivo. PMID- 12727386 TI - Laws, war, and public health. PMID- 12727388 TI - HIV/AIDS and people with disability. PMID- 12727387 TI - Imatinib for chronic myelogenous leukaemia: a 9 or 24 carat gold standard? PMID- 12727389 TI - FDA: untapped source of unpublished trials. PMID- 12727390 TI - Low-back pain in children. PMID- 12727391 TI - Chateau resveratrol. PMID- 12727392 TI - Mammography service screening and mortality in breast cancer patients: 20-year follow-up before and after introduction of screening. AB - BACKGROUND: The long term effect of mammographic service screening is not well established. We aimed to assess the long-term effect of mammographic screening on death from breast cancer, taking into account potential biases from self selection, changes in breast cancer incidence, and classification of cause of death. METHODS: We compared deaths from breast cancer diagnosed in the 20 years before screening was introduced (1958-77) with those from breast cancer diagnosed in the 20 years after the introduction of screening (1978-97) in two Swedish counties, in 210000 women aged 20-69 years. We also compared deaths from all cancers and from all causes in patients diagnosed with breast cancer in the 20 years before and after screening was introduced. In the analysis, data were stratified into age-groups invited for screening (40-69 years) and not invited (20-39 years), and by whether or not the women had actually received screening. We also analysed mortality for the 40-49-year age-group separately. FINDINGS: The unadjusted risk of death from breast cancer dropped significantly in the second screening period compared with the first in women aged 40-69 years (relative risk [RR] 0.77 [95% CI 0.7-0.85]; p<0.0001). No such decline was seen in 20-39 year olds. After adjustment for age, self-selection bias, and changes in breast-cancer incidence in the 40-69 years age-group, breast-cancer mortality was reduced in women who were screened (0.56; 0.49-0.64 p<0.0001), in those who were not screened (0.84 [0.71-0.99]; p=0.03), and in screened and unscreened women combined (0.59 [0.53-0.66]; p<0.0001). After adjustment for age, self-selection bias, and changes in incidence in the 40-49-year age-group, deaths from breast cancer fell significantly in those who were screened (0.52 [0.4-0.67]; p<0.0001); and in all women, screened and unscreened combined (0.55 [0.44-0.7] p<0.0001) but not in unscreened women (p=0.2). In both 40-69-year and 40-49-year age-groups, reductions in deaths from all cancers and from all-causes in women with breast cancer were consistent with these results. INTERPRETATION: Taking account of potential biases, changes in clinical practice and changes in the incidence of breast cancer, mammography screening is contributing to substantial reductions in breast cancer mortality in these two Swedish counties. PMID- 12727393 TI - Initiation of population-based mammography screening in Dutch municipalities and effect on breast-cancer mortality: a systematic review. AB - BACKGROUND: More than a decade ago, a mammography screening programme for women aged 50-69 years was initiated in the Netherlands. Our aim was to assess the effect of this programme on breast-cancer mortality rates. METHODS: We examined data for 27948 women who died of breast-cancer aged 55-74 years between 1980 and 1999 (30560 cases until 2001). We grouped individuals into 93 clusters, depending on where they lived, and analysed data by use of national population statistics. We analysed time trends in breast-cancer mortality, adjusting for gradual implementations at municipality level, taking as year 0 the month and year in which screening began in a particular municipality. We used a Poisson regression model to estimate the time at which the trend started to turn. We assessed indirectly whether this turning point was related to initiation of screening or adjuvant systemic therapy in four clusters defined according to when screening was implemented. FINDINGS: Compared with rates in 1986-88, breast-cancer mortality rates in women aged 55-74 years fell significantly in 1997 and subsequent years as predicted, reaching -19.9% in 2001. Mortality rates had been increasing by an annual 0.3% until screening was introduced; thereafter we noted a decline of 1.7% per year (95% CI 2.39-0.96) in women aged 55-74 years and of 1.2% in those aged 45-54 (2.40 to 0.07). The turning point in mortality trends arose at around year 0. Adjuvant systemic therapy is unlikely to be the cause of this turning point, since the mortality rates continued to rise up to 1 year after implementation in municipalities where screening began after 1995. INTERPRETATION: Routine mammography screening can reduce breast-cancer mortality rates in women aged 55-74 years. PMID- 12727394 TI - Two hard lessons. PMID- 12727395 TI - Effect of community-based promotion of exclusive breastfeeding on diarrhoeal illness and growth: a cluster randomised controlled trial. AB - BACKGROUND: Exclusive breastfeeding is recommended until age 6 months. We assessed the feasibility, effectiveness, and safety of an educational intervention to promote exclusive breastfeeding for this length of time in India. METHODS: We developed the intervention through formative research, pair-matched eight communities on their baseline characteristics, and randomised one of each pair to receive the intervention and the other to no specific intervention. We trained health and nutrition workers in the intervention communities to counsel mothers for exclusive breastfeeding at multiple opportunities. We enrolled 1115 infants born in the 9 months after training-552 in the intervention and 473 in the control communities. Feeding at age 3 months, and anthropometry and of diarrhoea prevalence at age 3 months and 6 months were assessed. All analyses were by intention to treat. FINDINGS: We assessed 483 and 412 individuals at 3 months in the intervention and control groups, respectively, and 468 and 412 at 6 months. At 3 months, exclusive breastfeeding rates were 79% (381) in the intervention and 48% (197) in the control communities (odds ratio 4.02, 95% CI 3.01-5.38, p<0.0001). The 7-day diarrhoea prevalence was lower in the intervention than in the control communities at 3 months (0.64, 0.44-0.95, p=0.028) and 6 months (0.85, 0.72-0.99, p=0.04). The mean weights and lengths, and the proportion with weight-for-height or height-for-age Z scores of 2 or less, at age 3 months and 6 months did not differ much between groups. Intervention effect on exclusive breastfeeding, diarrhoeal morbidity, and anthropometry at age 6 months in the low-birthweight subgroup was similar to that for all births. INTERPRETATION: Promotion of exclusive breastfeeding until age 6 months in a developing country through existing primary health-care services is feasible, reduces the risk of diarrhoea, and does not lead to growth faltering. PMID- 12727396 TI - Genetic regulation of fibrin structure and function: complex gene-environment interactions may modulate vascular risk. AB - BACKGROUND: Polymorphisms in the fibrinogen and factor XIII genes are associated with atherothrombotic risk, but clinical studies have produced inconsistent results and laboratory studies have not explained these findings. We aimed to investigate interactions between polymorphisms in the factor XIII and fibrinogen genes, fibrinogen concentrations, and other cardiovascular risk factors in relation to fibrin structure and function. METHODS: We used permeation analysis and electron microscopy to investigate interactions between fibrin structure, factor XIII Val34Leu, fibrinogen Aalpha Thr312Ala, fibrinogen Bbeta Arg448Lys, and fibrinogen concentrations in plasma and purified systems. FINDINGS: Increased fibrinogen concentrations were associated with decreases in permeability, with tighter clot structures in the presence of factor XIII 34Val alleles compared with those in the presence of 34Leu alleles. Findings were confirmed by scanning electron microscopy of fibrin. Similar changes in permeability were noted for Aalpha fibrinogen 312Ala compared with that for 312Thr. INTERPRETATION: Our results show interactions between coding polymorphisms in fibrinogen and factor XIII and fibrinogen concentrations that modify fibrin and explain the apparent paradox between epidemiological studies of factor XIII 34Leu and reported in vitro effects on fibrin structure and function. We suggest a potential complexity of gene-gene and gene-environment interactions in determining cardiovascular risk. PMID- 12727397 TI - ECG artifact due to deep brain stimulation. PMID- 12727398 TI - Ventricular fibrillation in a young Asian man. PMID- 12727399 TI - D,L-3-hydroxybutyrate treatment of multiple acyl-CoA dehydrogenase deficiency (MADD). AB - Cardiomyopathy and leukodystrophy are life-threatening complications of multiple acyl-CoA dehydrogenase deficiency (MADD). A 2-year-old boy with this disorder developed rapidly progressive leukodystrophy resulting in complete paralysis within 4 months. Within a week of starting sodium-D,L-3-hydroxybutyrate he had improved. After 2 years, neurological function returned, including walking independently, with progressive improvement of brain MRI. Two additional infants with MADD developed life-threatening cardiomyopathy unresponsive to conventional treatment. On sodium-D,L-3-hydroxybutyrate treatment their cardiac contractility showed progressive and sustained improvement. D,L-3-hydroxybutyrate is a therapeutic option for cerebral and cardiac complications in severe fatty acid oxidation defects. PMID- 12727400 TI - Control of encrustation and blockage of Foley catheters. AB - Urinary catheters often become encrusted and blocked by crystalline Proteus mirabilis biofilms. Results of experiments in a laboratory model of a Foley catheterised bladder infected with P mirabilis showed that when retention balloons were inflated with a solution of triclosan (10 g/L), the catheters drained freely for at least 7 days. Triclosan became impregnated throughout the silicone catheter material and completely inhibited the formation of crystalline biofilm, whereas catheters inflated with water became blocked in 24 h. Our observations suggest a way to control a common complication in patients with long term indwelling bladder catheters. PMID- 12727402 TI - China finally throws full weight behind efforts to contain SARS. PMID- 12727401 TI - Stapled versus excision haemorrhoidectomy: long-term follow up of a randomised controlled trial. AB - Advantages of the stapling procedure for haemorrhoids include reduced postoperative pain and shortened convalescence; however, there are few data with respect to functional and symptomatic outcome. At a dedicated clinic, we reviewed patients between Dec, 2001, and March, 2002, who had taken part in a randomised controlled trial undertaken at the unit in 1999, which compared outcomes after open or stapled haemorrhoidectomy. We noted the presence or absence of haemorrhoid specific symptoms, and assessed overall satisfaction, continence, and quality of life. Rigid sigmoidoscopy and an anorectal examination were also used to examine symptomatic recurrence and disease activity. At minimum follow-up of 33 months since surgery, both techniques seem to be equally effective. PMID- 12727403 TI - Media giant and foundation team up to fight HIV/AIDS. PMID- 12727404 TI - WHO's diet report prompts food industry backlash. PMID- 12727407 TI - USA revamps its HIV prevention efforts. PMID- 12727409 TI - Dutch veterinarian becomes first victim of avian influenza. PMID- 12727411 TI - Uganda leads Africa's charge against guinea worm disease. PMID- 12727412 TI - Pancreatitis. AB - In the past decade, our understanding of the genetic basis, pathogenesis, and natural history of pancreatitis has grown strikingly. In severe acute pancreatitis, intensive medical support and non-surgical intervention for complications keeps patients alive; surgical drainage (necrosectomy) is reserved for patients with infected necrosis for whom supportive measures have failed. Enteral feeding has largely replaced the parenteral route; controversy remains with respect to use of prophylactic antibiotics. Although gene therapy for chronic pancreatitis is years away, our understanding of the roles of gene mutations in hereditary and sporadic pancreatitis offers tantalising clues about the disorder's pathogenesis. The division between acute and chronic pancreatitis has always been blurred: now, genetics of the disorder suggest a continuous range of disease rather than two separate entities. With recognition of pancreatic intraepithelial neoplasia, we see that chronic pancreatitis is a premalignant disorder in some patients. Magnetic resonance cholangiopancreatography and endoscopic ultrasound are destined to replace endoscopic retrograde cholangiopancreatography for many diagnostic indications in pancreatic disease. PMID- 12727413 TI - New surgical treatments for menorrhagia. AB - CONTEXT: Hysterectomy is a common and effective treatment for menorrhagia but is associated with substantial post-operative convalescence time and morbidity. In the early 1990s endometrial resection or ablation became a well-established day case alternative for the surgical treatment of menorrhagia. Both endometrial resection and ablation require general anaesthesia, a high level of skill in hysteroscopic surgery, and can be long procedures. More recently, various new techniques have been developed that can be done in an outpatient setting under local anaesthesia and with a low risk of complications. The effectiveness of most new second-generation ablation technologies has not been confirmed in randomised trials and it is possible that these techniques will not prove to be as effective or as safe as originally thought. STARTING POINT: Massimiliano Pellicano and colleagues (Am J Obstet Gynecol 2002; 187: 545-50) compared a second-generation ablation technique, thermal destruction of the endometrium with a heated-water filled silicone balloon with hysteroscopic endometrial resection. 82 women were randomised and followed up for 2 years. Thermal destruction was quicker than hysteroscopic resection, and was associated with a similar level of postoperative satisfaction and reintervention rate. This study suggests that thermal destruction is as effective a technique as endometrial resection. WHERE NEXT? Many second-generation ablation techniques are now available. Some may prove more effective than others, but much larger studies are needed to address safety. The development of progestagen-releasing intrauterine devices, which provide effective treatment for menorrhagia and are also an effective and reversible form of contraception, may mean that the uptake of second-generation surgical ablation techniques is less widespread than some proponents of these new technologies suggest. PMID- 12727414 TI - Assessment of travellers who return home ill. AB - Every year, millions of people travel abroad, exposing themselves to various diseases. Advice on risk avoidance and on self-medication is not always successful; sometimes travellers return home ill or become unwell soon afterwards. There are many possible causes for such illnesses, and physicians should try to establish whether the disease is specifically associated with the recent journey. The approach to assessment of the ill traveller should make use not only of signs and symptoms, but also of geography and epidemiology. Travellers with fever need immediate attention to rule out serious and potentially life-threatening conditions. Faced with a difficult diagnosis, physicians should consult with experts in tropical and travel medicine. PMID- 12727415 TI - Shooting at ambulances in Israel: a cardiologist's viewpoint. PMID- 12727416 TI - Humanitarian propaganda. PMID- 12727417 TI - Relief operations in Iraq. PMID- 12727418 TI - New European Directive on clinical trials. PMID- 12727419 TI - Is primary care research a lost cause? PMID- 12727420 TI - Is primary care research a lost cause? PMID- 12727421 TI - Preparedness of London hospitals for a chemical weapons attack. PMID- 12727422 TI - The neglect of child neglect. PMID- 12727423 TI - The neglect of child neglect. PMID- 12727424 TI - UK Hip Trial. PMID- 12727427 TI - Report of the WHO Commission on Macroeconomics and Health. PMID- 12727428 TI - P2X7 polymorphism and chronic lymphocytic leukaemia. PMID- 12727430 TI - Extracampine hallucinations. PMID- 12727431 TI - Extracampine hallucinations. PMID- 12727432 TI - Laboratory preimplantation genetic diagnosis. PMID- 12727433 TI - Intact fetal cells in maternal plasma. PMID- 12727434 TI - How did Vasco da Gama sail for 16 weeks without developing scurvy? PMID- 12727435 TI - All in the family. PMID- 12727440 TI - Expression of MeCP2 in olfactory receptor neurons is developmentally regulated and occurs before synaptogenesis. AB - Rett syndrome, a neurodevelopmental disorder hypothesized to be due to defective neuronal maturation, is a result of mutations in the mecp2 gene encoding the transcriptional repressor methyl-CpG binding protein (MeCP2). We utilized the olfactory system, which displays postnatal neurogenesis, as a model to investigate MeCP2 expression during development and after injury. MeCP2 expression increased postnatally, localizing to mature olfactory receptor neurons (ORNs) and sustentacular supporting cells. The timing of MeCP2 expression was defined by using detergent ablation (to remove the ORNs) and unilateral olfactory bulbectomy (to remove the ORN target), both of which increase neurogenesis. MeCP2 expression in the ORNs reached prelesioning levels as cells matured after ablation, whereas expression was not completely restored after bulbectomy, in which functional synaptogenesis cannot occur. Thus, MeCP2 expression correlates with the maturational state of ORNs, and precedes synaptogenesis. Identifying the time window of MeCP2 expression should help further clarify the biological defects in Rett syndrome. PMID- 12727441 TI - Tamoxifen-inducible glia-specific Cre mice for somatic mutagenesis in oligodendrocytes and Schwann cells. AB - Inducible transgenesis provides a valuable technique for the analysis of gene function in vivo. We report the generation and characterization of mouse lines carrying glia lineage-specific transgenes expressing an improved variant of the tamoxifen-inducible Cre recombinase, CreERT2, where the recombinase is fused to a mutated ligand binding domain of the human estrogen receptor. Using a PLP-CreERT2 transgene, we have generated mice that show specific inducible Cre function, as analyzed by cross-breeding experiments into the Rosa26 Cre-LacZ reporter line, in developing and adult Schwann cells, in mature myelinating oligodendrocytes, and in undifferentiated NG2-positive oligodendrocyte precursors in the adult. Using a P0Cx-CreERT2 transgene, we have also established mouse lines with inducible Cre function specifically in the Schwann cell lineage. These tamoxifen-inducible CreERT2 lines will allow detailed spatiotemporally controlled analysis of gene functions in loxP-based conditional mutant mice in both developing and adult Schwann cells and in the oligodendrocyte lineage. PMID- 12727443 TI - Dynamics of motor nerve terminal remodeling unveiled using SNARE-cleaving botulinum toxins: the extent and duration are dictated by the sites of SNAP-25 truncation. AB - Nerve sprouts emerge from motor nerve terminals following blockade of exo endocytosis for more than 3 days by botulinum neurotoxin (BoNT), and form functional synapses, albeit temporary. Upon restoration of synaptic activity to the parent terminal 7 and 90 days after exposure to BoNT/F or A respectively, a concomitant retraction of the outgrowths was observed. BoNT/E caused short-term neuroparalysis, and dramatically accelerated the recovery of BoNT/A-paralyzed muscle by further truncation of SNAP-25 and its replenishment with functional full-length SNARE. The removal of 9 C-terminal residues from SNAP-25 by BoNT/A leads to persistence of the inhibitory product due to the formation of a nonproductive SNARE complex(es) at release sites, whereas deletion of a further 17 amino acids permits replenishment and a speedy recovery. PMID- 12727444 TI - Distribution of p120 catenin during rat brain development: potential role in regulation of cadherin-mediated adhesion and actin cytoskeleton organization. AB - p120 catenin (p120ctn) is implicated in the regulation of cadherin-mediated adhesion and actin cytoskeleton remodeling. The interaction of cytoplasmic p120ctn with the guanine exchange factor Vav2 is one of the signaling pathways implicated in cytoskeleton dynamics. We show here that p120ctn is regulated during rat brain development and is distributed at the membrane and within the cytoplasm where it associates with N-cadherin and Vav2, respectively. p120ctn shifts progressively from an axonal expression to a punctuate staining localized to a subset of synapses. In cultured hippocampal neurons, p120ctn redistributes from growth cones to synapses, where it partly colocalizes with N-cadherin or Vav2 and filamentous actin. In the adult forebrain, we show that p120ctn and Vav2 are highly expressed by neuroblasts migrating from the lateral subventricular zone to the olfactory bulb. The dynamic expression pattern of p120ctn and the biochemical evidences of its association with N-cadherin and Vav2 strongly suggest that p120ctn plays a major role in neuronal migration, neurite outgrowth and synapse formation, and plasticity. PMID- 12727442 TI - Requirement of the orphan nuclear receptor SF-1 in terminal differentiation of ventromedial hypothalamic neurons. AB - The ventromedial hypothalamic nucleus (VMN) is known to mediate autonomic responses in feeding and reproductive behaviors. To date, the most definitive molecular marker for the VMN is the orphan nuclear receptor steroidogenic factor 1 (SF-1). However, it is unclear whether SF-1 functions in the VMN as it does in peripheral endocrine organ development where loss of SF-1 results in organ agenesis due to apoptosis. Here, we provide evidence that SF-1 has a distinct role in later stages of VMN development by demonstrating the persistence of VMN precursors, the misexpression of an early marker (NKX2-1) concomitant with the absence of a late marker (BDNF neurotrophin), and the complete loss of projections to the bed nucleus of stria terminalis and the amygdala in sf-1 null mice. Our findings demonstrate that SF-1 is required for terminal differentiation of the VMN and suggest that transcriptional targets of SF-1 mediate normal circuitry between the hypothalamus and limbic structures in the telencephalon. PMID- 12727445 TI - Interleukin-1 beta protects neurons via the interleukin-1 (IL-1) receptor mediated Akt pathway and by IL-1 receptor-independent decrease of transmembrane currents in vivo. AB - Recently, we have demonstrated that tumor necrosis factor-alpha (TNF-alpha) rescues retinal ganglion cells (RGCs) from retrograde cell death in vivo after axotomy of the optic nerve. The mechanism of RGC rescue was dependent on TNF receptor I-mediated potassium current reduction and consecutive activation of the phosphatidylinositol 3-kinase (PI3-K)/Akt pathway. Here, we present evidence that interleukin-1 beta (IL-1 beta) also promotes RGC survival, but shows distinct differences with respect to its neuroprotective mechanisms. Using whole-cell and outside-out patch-clamp techniques, we observed that IL-1 beta decreased both inward sodium current amplitudes and outward potassium current amplitudes. Counteracting these effects by sodium or potassium channel opening inhibited the survival-promoting effects of this cytokine. IL-1 beta-induced current reduction could not be abolished by the interleukin-1 receptor antagonist, indicating that the electrophysiological effects of IL-1 beta are independent of interleukin-1 receptor I (IL-1RI) activation. Western blot analysis revealed an IL-1 beta induced IL-1RI-dependent upregulation of phospho-Akt. Antagonism of the survival promoting effects of IL-1 beta by PI3-K inhibition revealed the functional relevance of the PI3-K/Akt pathway in IL-1 beta-induced signal transduction in vivo. PMID- 12727446 TI - SHH and FGF8 play distinct roles during development of noradrenergic neurons in the locus coeruleus of the zebrafish. AB - Several signaling pathways have been implicated in the development of dopaminergic and serotonergic neurons. Here, we analyzed the formation of noradrenergic (NAergic) cells in the locus coeruleus (LC) of zebrafish. In the sonic hedgehog (shh) mutant, cells positive for tyrosine hydroxylase, a marker for putative NAergic cells in the LC were reduced. Similarly, the inhibition of translation of all hh genes and the perturbation of Shh signaling by forskolin resulted in a decrease in the number of cells. Conversely, when SHH was overexpressed, an increase in number was observed. Thus, Shh is involved in maintaining the appropriate number of cells in the LC. While elevated levels of bone morphogenetic protein 4 (BMP4) did not attenuate tyrosine hydroxylase positive cells, exogenous fibroblast growth factor 8 (FGF8) rescued NAergic neurons in the acerebellar (ace) mutant, providing direct in vivo evidence that Fgf8 is required for the induction of NAergic neurons in the LC. PMID- 12727448 TI - The Dlx5 homeodomain gene is essential for olfactory development and connectivity in the mouse. AB - The distalless-related homeogene Dlx5 is expressed in the olfactory placodes and derived tissues and in the anterior-basal forebrain. We investigated the role of Dlx5 in olfactory development. In Dlx5(-/-) mice, the olfactory bulbs (OBs) lack glomeruli, exhibit disorganized cellular layers, and show reduced numbers of TH- and GAD67-positive neurons. The olfactory epithelium in Dlx5(-/-) mice is composed of olfactory receptor neurons (ORNs) that appear identical to wild-type ORNs, but their axons fail to contact the OBs. We transplanted Dlx5(-/-) OBs into a wild-type newborn mouse; wild-type ORN axons enter the mutant OB and form glomeruli, but cannot rescue the lamination defect or the expression of TH and GAD67. Thus, the absence of Dlx5 in the OB does not per se prevent ORN axon ingrowth. In conclusion, Dlx5 plays major roles in the connectivity of ORN axons and in the differentiation of OB interneurons. PMID- 12727449 TI - Identification of teleost Thy-1 and association with the microdomain/lipid raft reggie proteins in regenerating CNS axons. AB - During regeneration, retinal ganglion cell axons in fish upregulate a cell surface protein that is recognized by the monoclonal antibody (mAB) M802. M802 antigen appeared to be linked to the intracellular, membrane-associated lipid raft/microdomain proteins reggie-1 and reggie-2 that were previously shown to be reexpressed in axon-regenerating neurons [Development 124 (1997), 577]. Here, we report the isolation of the M802 antigen and its identification as the teleost homolog of mammalian Thy-1. Fish Thy-1 is detected in the same detergent insoluble lipid raft fractions from a fibroblast cell line and from axon regenerating retinae as reggie-1 and 2. Importantly, mAB M802 coimmunoprecipitates reggie-1 and 2 from this lipid raft fraction, implying that fish Thy-1 and reggies interact. This correlates with their colocalization in growing cell processes after M802 antigen/Thy-1 activation with mAB M802. These findings suggest a role of clustered M802 antigen/Thy-1 in reggie raft microdomains for cell growth and axon regeneration. PMID- 12727447 TI - Transcription factor expression and cellular redox in immature oligodendrocyte cell death: effect of Bcl-2. AB - Multiple sclerosis (MS) is characterized by the progressive damage or loss of oligodendrocytes. In an effort to better understand the causes of oligodendrocyte destruction in MS plaques, we treated immature oligodendrocytes with glucose oxidase, ceramide, or brefeldin A. These treatments model the different mechanisms by which oligodendrocytes are thought to die. We report that the AP-1 and Egr-1 transcription factors are induced within an hour of treatment. Of the AP-1 proteins studied, c-Jun was expressed at the highest level, followed by JunD, c-Fos, and Fra-2, although different treatments induced slightly different levels of expression. Bcl-2 overexpression protects against all treatments, to differing degrees. Although Bcl-2 did not have a dramatic effect on AP-1 or Egr-1 induction within the first 3 h, it caused a lowering of steady-state redox levels with a concomitant increase in cellular glutathione. We propose that the lowering of cellular redox and the upregulation of glutathione are responsible in part for the protective properties of Bcl-2. PMID- 12727450 TI - Statistics of trinucleotides in coding sequences and evolution. AB - The aim of this paper is to give measurements indicative of evolutional stages of the species. Two types of statistics of trinucleotides in coding regions are analysed for 27 species. The first one is the codon space, the nucleotide ratio for each of the three codon positions. We apply principal component analysis on this space and extract two principal components faithfully describing the original distribution of the codon space. The first principal component corresponds to the GC content. The second principal component classifies the species into three evolutional groups, Archaea, Bacteria and Eukaryota. The second statistics is the real and theoretical frequency of amino acids. The real frequency of an amino acid in a coding sequence is its frequency in the translated protein. The theoretical frequency is the expected frequency calculated from the ratio of nucleotides. We introduce the discrepancy between these two frequencies as an index of non-randomness of nucleotides in the sequence. This index of non-randomness divides the species into two groups: eukaryotes having smaller non-randomness (i.e. being more random) and prokaryotes having higher non-randomness. PMID- 12727451 TI - Effects of correlated and synchronized stochastic inputs to leaky integrator neuronal model. AB - We present an analysis of neuronal model behaviour with correlated synaptic inputs including the cases that correlated inputs are equivalent to exactly synchronized inputs and correlated inputs are not equivalent to exactly synchronized inputs. For the former case, it is found that the fully (synaptically) correlated inputs assumption (see Section 1 for definition), which is used in most, if not all, theoretical and experimental work in the past few years, results in a waste of resources and might be an unrealistic assumption; with an exactly balanced excitatory and inhibitory, and synaptically correlated input, the integrate-and-fire model simply behaves as a synchrony detector in certain parameter regions; the well-known diffusion model, upon which most theoretical work is based, fails to approximate the model with synaptically correlated Poisson inputs. A novel way to approximate synaptically correlated Poisson inputs is then presented;an optimization principle on neuronal models with partially (synaptically) correlated inputs is proposed, which enables us to predict microscopic structures in neuronal systems. For the latter case,with tightly synchronized inputs (see Section 1 for definition), the model behaviour depends on its integration time of input signals and could exhibit bursting discharge.for loosely synchronized inputs, we found that correlated inputs are equivalent to the post-spike voltage reset mechanism proposed in the literature. PMID- 12727452 TI - Neurophysical theory of coherence and correlations of electroencephalographic and electrocorticographic signals. AB - Correlation and coherence functions of electroencephalographic signals are calculated using a recent continuum theory that has previously yielded excellent agreement with observations of electroencephalographic spectra. The predicted properties of these functions are found to be in semiquantitative agreement with observations for parameters consistent with those used in previous studies of spectra. The corresponding results for electrocorticographic signals point to an additional contribution at characteristic scales of around 6mm, as has been previously inferred, and are consistent with a crossover to the long-range behavior seen in scalp data. Analysis within the framework of the model enables constraints on the relative strengths of the two contributions to be inferred, and makes it plausible that the short-range component reflects the point-spread function of external stimuli and corticothalamic feedback reaching the cortex. PMID- 12727453 TI - Block structure and stability of the genetic code. AB - It is known that different codons may be unified into larger groups related to the hierarchical structure, approximate hidden symmetries, and evolutionary origin of the universal genetic code. Using a simplified evolutionary motivated two-letter version of genetic code, the general principles of the most stable coding are discussed. By the complete enumeration in such a reduced code it is strictly proved that the maximum stability with respect to point mutations and shifts in the reading frame needs the fixation of the middle letters within codons in groups with different physico-chemical properties, thus, explaining a key feature of the universal genetic code. The translational stability of the genetic code is studied by the mapping of code onto de Bruijn graph providing both the compact visual representation of mutual relationships between different codons as well as between codons and protein coding DNA sequence and a powerful tool for the investigation of stability of protein coding. Then, the results are extended to four-letter codes. As is shown, the universal genetic code obeys mainly the principles of optimal coding. These results demonstrate the hierarchical character of optimization of universal genetic code with strictly optimal coding being evolved at the earliest stages of molecular evolution. Finally, the universal genetic code is compared with the other natural variants of genetic codes. PMID- 12727454 TI - On parametric evenness measures. AB - The degree to which abundances are divided equitably among community species or evenness is a basic property of any biological community. Several evenness indices have thus far been proposed in ecological literature. However, despite its vast potential applicability in ecological research, none seems to be generally preferred. Furthermore, only very few parametric evenness families have thus far been proposed. While traditional evenness indices supply point descriptions of community evenness, according to a parametric evenness family E(alpha), there is a continuum of possible evenness measures that differ in their sensitivity to changes in the relative abundances of dominant and rare species as a function of the parameter alpha. In this review, I first summarize the basic requirements that a parametric evenness measure should meet to adequately behave in ecological studies. Next, I discuss the major drawbacks of these requirements and propose some alternative solutions. PMID- 12727455 TI - Evolving protein interaction networks through gene duplication. AB - The topology of the proteome map revealed by recent large-scale hybridization methods has shown that the distribution of protein-protein interactions is highly heterogeneous, with many proteins having few edges while a few of them are heavily connected. This particular topology is shared by other cellular networks, such as metabolic pathways, and it has been suggested to be responsible for the high mutational homeostasis displayed by the genome of some organisms. In this paper we explore a recent model of proteome evolution that has been shown to reproduce many of the features displayed by its real counterparts. The model is based on gene duplication plus re-wiring of the newly created genes. The statistical features displayed by the proteome of well-known organisms are reproduced and suggest that the overall topology of the protein maps naturally emerges from the two leading mechanisms considered by the model. PMID- 12727456 TI - Biomechanical models of hyphal growth in actinomycetes. AB - The tip growth of filamentary actinomycetes is investigated within the framework of large deformation membrane theory in which the cell wall is represented as a growing elastic membrane with geometry-dependent elastic properties. The model exhibits realistic hyphal shapes and indicates a self-similar tip growth mechanism consistent with that observed in experiments. It also demonstrates a simple mechanism for hyphal swelling and beading that is observed in the presence of a lysing agent. PMID- 12727457 TI - First-order differential equation models with estimable parameters as functions of environmental variables and their application to a study of vascular development in young hybrid aspen stems. AB - A novel method is described for solving systems of differential equations pertaining to organism development, where this development is assumed to be directly influenced by fluctuation in measurable environmental variables. The system parameters are written as functions of these variables and, because these functions involve the accumulation of "environment time" (e.g., "day-degrees"), the system is therefore regulated by the prevailing environmental conditions. This method contrasts with the more usual descriptions of development along a time-line. The parameters of the differential equations involved are estimated by modelling data, which show evidence of changes in the dependent variable(s), i.e. the components of the system. They are expressed in terms of their response to continuous fluctuations in one or more independent, environmental variables. Accumulated thermal time (including day-degrees) or more complex units may be derived by using either linear or nonlinear functions. Critical environmental parameters such as the basal thresholds of a given developmental process or parameters describing a nonlinear relationship with the environment may then be estimated. This paper develops the methodology of this environmentally driven approach to describing organism development in general terms, and gives a specific example of its application with reference to the cellular development within the secondary vascular tissues in the stems of young hybrid aspen trees. PMID- 12727458 TI - Evolutionary stability in Lotka-Volterra systems. AB - The Lotka-Volterra model of population ecology, which assumes all individuals in each species behave identically, is combined with the behavioral evolution model of evolutionary game theory. In the resultant monomorphic situation, conditions for the stability of the resident Lotka-Volterra system, when perturbed by a mutant phenotype in each species, are analysed. We develop an evolutionary ecology stability concept, called a monomorphic evolutionarily stable ecological equilibrium, which contains as a special case the original definition by Maynard Smith of an evolutionarily stable strategy for a single species. Heuristically, the concept asserts that the resident ecological system must be stable as well as the phenotypic evolution on the "stationary density surface". The conditions are also shown to be central to analyse stability issues in the polymorphic model that allows arbitrarily many phenotypes in each species, especially when the number of species is small. The mathematical techniques are from the theory of dynamical systems, including linearization, centre manifolds and Molchanov's Theorem. PMID- 12727459 TI - Simulating convergent extension by way of anisotropic differential adhesion. AB - Simulations using the Extended Potts Model suggest that anisotropic differential adhesion can account for convergent extension, as observed during embryonic development of the frog Xenopus laevis for example. During gastrulation in these frogs, convergent extension produces longitudinal tissue growth from latitudinal elongation and migration of aligned constituent cells. The Extended Potts Model employs clustered points on a grid to represent subdivided cells with probabilistic displacement of cell boundaries such that small changes in energy drive gradual tissue development. For modeling convergent extension, simulations include anisotropic differential adhesion: the degree of attachment between adjacent elongated cells depends on their relative orientation. Without considering additional mechanisms, simulations based on anisotropic differential adhesion reproduce the hallmark stages of convergent extension in the correct sequence, with random fluctuations as sufficient impetus for cell reorganization. PMID- 12727460 TI - Autopoietic and (M,R) systems. AB - From the many attempts to produce a conceptual framework for the organization of living systems, the notions of (M,R) systems and Autopoiesis stand out for their rigor, their presupposition of the circularity of metabolism, and the new epistemologies that they imply. From their inceptions, these two notions have been essentially disconnected because each has defined its own language and tools. Here we demonstrate the existence of a deep conceptual link between (M,R) systems and Autopoietic systems. This relationship permits us to posit that Autopoietic systems, which have been advanced as capturing the central aspects of living systems, are a subset of (M,R) systems. This result, in conjunction with previous theorems proved by Rosen, can be used to outline a demonstration that the operation of Autopoietic systems cannot be simulated by Turing machines. This powerful result shows the potential of linking these two models. Finally, we suggest that the formalism of (M,R) systems could be used to model the circularity of metabolism. PMID- 12727461 TI - Expansion of the neocerebellum in Hominoidea. AB - Technological and conceptual breakthroughs have led to more serious consideration of the cerebellum as an essential element in cognition. Recent studies show the lateral cerebellum, seat of the neocerebellum, to be most active in cognitive tasks. An examination of the relative volumes of the cerebellar hemispheres in anthropoids would reveal whether some groups show greater neocerebellar development through hemispheric expansion beyond expected allometry, implying a greater contribution of the lateral hemispheres to cognition. This study expands the existing data on primate brain and brain part volumes by incorporating data from both magnetic resonance scans and histological sections for a total sample size of 97 specimens, including 42 apes, 14 humans and 41 monkeys. The resulting volumes of whole brain, cerebellum, vermis, and hemisphere enable a reliable linear regression contrast between hominoids and monkeys, and demonstrate a striking increase in the lateral cerebellum in hominoids. The uniformity of the grade shift suggests that this increase took place in the common ancestor to the hominoids. The importance of the neocerebellum in visual-spatial skills, planning of complex movements, procedural learning, attention switching, and sensory discrimination in manipulation would facilitate the adaptation of these early hominoids to frugivory and suspensory feeding. PMID- 12727462 TI - Intrinsic qualities of primate bones as predictors of skeletal element representation in modern and fossil carnivore feeding assemblages. AB - Plio-Pleistocene faunal assemblages from Swartkrans Cave (South Africa) preserve large numbers of primate remains. Brain, C.K., 1981. The Hunters or the Hunted? An Introduction to African Cave Taphonomy. University of Chicago Press, Chicago suggested that these primate subassemblages might have resulted from a focus by carnivores on primate predation and bone accumulation. Brain's hypothesis prompted us to investigate, in a previous study, this taphonomic issue as it relates to density-mediated destruction of primate bones (J. Archaeol. Sci. 29, 2002, 883). Here we extend our investigation of Brain's hypothesis by examining additional intrinsic qualities of baboon bones and their role as mediators of skeletal element representation in carnivore-created assemblages. Using three modern adult baboon skeletons, we collected data on four intrinsic bone qualities (bulk bone mineral density, maximum length, volume, and cross-sectional area) for approximately 81 bones per baboon skeleton. We investigated the relationship between these intrinsic bone qualities and a measure of skeletal part representation (the percentage minimum animal unit) for baboon bones in carnivore refuse and scat assemblages. Refuse assemblages consist of baboon bones not ingested during ten separate experimental feeding episodes in which individual baboon carcasses were fed to individual captive leopards and a spotted hyena. Scat assemblages consist of those baboon bones recovered in carnivore regurgitations and feces resulting from the feeding episodes. In refuse assemblages, volume (i.e., size) was consistently the best predictor of element representation, while cross-sectional area was the poorest predictor in the leopard refuse assemblage and bulk bone mineral density (i.e., a measure of the proportion of cortical to trabecular bone) was the poorest predictor in the hyena refuse assemblage. In light of previous documentation of carnivore-induced density-mediated destruction to bone assemblages, we interpret the current findings as suggestive of the secondary importance of bulk bone mineral density to other intrinsic qualities of skeletal elements (e.g., size, maximum dimension, and average cross-sectional area). It is only when skeletal elements are too large for consumption (e.g., many long bones) that they are fragmented following intra-element patterns of density-mediated carnivore destruction. There appears to be a size threshold beneath which bulk bone mineral density contributes little to mediating carnivore destruction of carcasses. Thus, depending on body size of the predator, body size of the prey, and specific size of the element, bulk bone mineral density may play little or no role of primary importance in mediating the destruction of skeletal elements. We compare patterns in modern comparative assemblages to patterns in primate fossil assemblages from Swartkrans. One of the fossil assemblages, Swartkrans Member 1, Hanging Remnant, most closely approximates a hyena (possibly refuse) assemblage pattern, while the Swartkrans Member 2 assemblage most closely approximates a leopard (possibly scat) assemblage pattern. The Swartkrans Member 1, Lower Bank, assemblage does not closely approximate any of our modern comparative assemblage patterns. PMID- 12727463 TI - An integrated approach to taphonomy and faunal change in the Shungura formation (Ethiopia) and its implication for hominid evolution. AB - Environmental and faunal changes through time have been recorded for many African Plio-Pleistocene sites. Fossil evidence suggests that there is a continuous, if not uniform, transformation of the fauna and flora from the Pliocene through the end of Pleistocene. However, discerning major biotic turnovers and linking them to global and regional climatic changes have been complicated by many factors, notably taphonomy and discontinuity of the fossil evidence, notwithstanding the considerable work of some researchers (e.g., Vrba, E.S., 1988. Late Pliocene climatic events and hominid evolution, in: Grine, F. (Ed.), Evolutionary History of the "Robust" Australopithecines. De Gruyter, New York, pp. 405-426, Vrba, E.S., 1995. The fossil record of African (Mammalia, Bovidae) in relation to human evolution and paleoclimate, in: Vrba, E.S., Denton, G.H., Partridge, T.C., Burkle, L.H. (Eds.), Paleoclimate and Evolution, with Emphasis on Human Origins. Yale University Press, New Haven, pp. 385-424). A sample of over 22,000 fossils collected by the French Omo Expedition, from the Shungura Formation of Ethiopia, was analyzed using an integrated approach to investigate taphonomic and faunal change patterns. The following results are obtained: (1) Univariate and multivariate studies support continuous faunal change from Member A through Member G of the Shungura sequence; (2) Correspondence analysis (CA) on extant bovids in African game parks shows that bovid tribes and genera are generally characterized by habitat specificity; (3) Taphonomic studies demonstrate that the relative abundance of different skeletal elements varies according to depositional environment; (4) CA on 73 localities of the Shungura Formation and 19 mammalian taxa points to a major faunal change around the base of Member G dated to ca. 2.3 Ma. This transformation is characterized by a change to open and edaphic grassland as a dominant type of environment; (5) This major faunal change correlates in time with the appearance of A. boisei. It is tentatively suggested that this major biome change is associated with an anagenetic speciation from A. aethiopicus to A. boisei. PMID- 12727464 TI - Palaeopathological and variant conditions of the Homo heidelbergensis type specimen (Mauer, Germany). AB - Although early Homo specimens are now known from a number of African, Asian and European Middle Pleistocene sites, the taxon Homo heidelbergensis was initially introduced for the Mauer jaw recovered in 1907. Fossil hominids from the earlier Middle Pleistocene of Europe are very rare and the Mauer mandible is generally accepted as one of the most ancient, with an age of approximately 700 kyr. A new preparation of the mandible was conducted in 1996 and gave rise to the detailed palaeopathological examination which is presented here. Based on comparative analyses, the extreme breadth of the mandibular ramus and its flat intercondylar incision, in conjunction with the flattening and broadening of the coronoid process tip, results either from an idiosyncratic pattern of the course and insertion of the temporalis muscle on the coronoid process or from the temporalis possessing an accessory head. The incidence of periodontal pocketing, together with a vertical reduction of the alveolar margin to approximately 3.00 mm, and a slight protuberance formed in vicinity of the right M(2)can safely be interpreted as pathognomonic indications of periodontal disease. The short distance between the enamel-dentine junction of the teeth and the horizontal alveolar margins could either be an inherited variant or may result from incipient osteoporosis. In addition, an arthrotic condition with slight osteophytic peripheral exostoses and an arthrolit (i.e. an articular calculus or "joint mouse") on the left condylus articularisand a depression in the medial part of the left mandibular condyle extending into the inferior part of the ramus are present. These features are indicative of a trauma-induced osteochondrosis dissecans. The diagnosis therefore suggests that the observed depression results from a well-healed fracture. This traumatic event illustrates the demanding living conditions endured by humans during the European Middle Pleistocene. The variations and pathological conditions observed in Mauer do not question the mandible's role as type specimen for the taxon Homo heidelbergensis. PMID- 12727465 TI - Non-occlusal dental microwear variability in a sample of Middle and Late Pleistocene human populations from Europe and the Near East. AB - Non-occlusal, buccal tooth microwear variability has been studied in 68 fossil humans from Europe and the Near East. The microwear patterns observed suggest that a major shift in human dietary habits and food processing techniques might have taken place in the transition from the Middle to the Late Pleistocene populations. Differences in microwear density, average length, and orientation of striations indicate that Middle Pleistocene humans had more abrasive dietary habits than Late Pleistocene populations. Both dietary and cultural factors might be responsible for the differences observed. In addition, the Middle Paleolithic Neanderthal specimens studied show a highly heterogeneous pattern of microwear when compared to the other samples considered, which is inconsistent with a hypothesis of all Neanderthals having a strictly carnivorous diet. The high density of striations observed in the buccal surfaces of several Neanderthal teeth might be indicative of the inclusion of plant foods in their diet. The buccal microwear variability observed in the Neanderthals is compatible with an overall exploitation of both plant and meat foods on the basis of food availability. A preliminary analysis of the relationship between buccal microwear density and climatic conditions prevailing in Europe during the Late Pleistocene has been attempted. Cold climatic conditions, as indicated by oxygen isotope stage data, seem to be responsible for higher densities of microwear features, whereas warmer periods could correspond to a reduced pattern of scratch density. Such a relationship would be indicative of less abrasive dietary habits, perhaps more meat dependent, during warmer periods. PMID- 12727466 TI - New fossil primate from Chile. PMID- 12727467 TI - Primates in caves: two new reports of Papio spp. PMID- 12727469 TI - Blockade of intragraft IL-2 receptor-alpha by basiliximab is insufficient to prevent activation of liver graft infiltrating cells. AB - Treatment with basiliximab, a CD25 (Interleukin-2 receptor-alpha; IL-2Ralpha) blocking human-murine chimeric antibody, reduces the incidence of acute rejections after organ transplantation, but rejection is not completely prevented. We investigated whether rejections during basiliximab treatment were due to insufficient exposure to the antibody or to incomplete blockade of intragraft CD25, and whether CD25-blockade affected activation of liver transplant infiltrating cells. Twenty-seven basiliximab-treated liver transplant recipients and seven patients not treated with basiliximab, from which post transplant liver biopsies were available, were retrospectively selected for this study. Rejectors among the basiliximab-treated patients (n=11) had not cleared basiliximab from their blood at a faster rate than non-rejectors (n=16). Rejections within the period of saturating serum basiliximab concentrations were in most cases not due to insufficient intragraft CD25-blockade: in eight out of eleven rejection biopsies obtained during this period CD25 on graft-infiltrating cells was completely coated by basiliximab. Despite CD25 blockade, portal infiltrates in these biopsies showed similar expression of the proliferation associated antigen Ki-67 and of the cytotoxic effector molecules granzyme A and B as those in rejection biopsies obtained from patients not treated with basiliximab. In conclusion, basiliximab treatment of liver transplant patients results in blockade of intragraft CD25, but nevertheless cells in rejection infiltrates are strongly activated, probably by a mechanism bypassing the IL 2Ralpha. PMID- 12727470 TI - Genetic modification of cold-preserved renal grafts using HSP70 or bcl-2 HVJ liposome method. AB - BACKGROUND: We tested the hypothesis that the best time for genetic modification is while the cell viability of the graft is reduced for long-term preservation. The hemagglutinating virus of Japan (HVJ)-liposome method, a nonviral gene transfer technique, was used with a luciferase gene to test the efficacy of protein induction under the critical preservation time. Furthermore, we tested this genetic modification with heat shock protein (HSP) 70 or bcl-2 genes to prevent primary nonfunction (PNF) after long-term preservation. METHODS: Orthotopic rat renal transplantation (RT) was performed using the cuff technique in the syngeneic combination of Lew (major histocompatible complex, haplotype: RT1(l)). Rat kidney grafts were preserved for 24 or 48 h in University of Wisconsin (UW) or Ringer's lactate solution using HVJ method with the luciferase gene. Rats with gene-transfected kidneys were re-laparotomized 48 h after transplantation to estimate the lack of arterial flow in the graft and killed for histological evaluation of the degree of PNF luciferase intensity assay. Then, two functional genes (HSP70 or bcl-2) were tested for the occurrence of PNF and histological and immunohistochemical analysis of the grafted kidneys preserved for 48 h in the UW solution. RESULTS: In the kidneys preserved for 24 h, 50% of the Ringer's lactate group had PNF; but all of the UW group had sufficient blood flow. The graft viability was well corrected by the degree of luciferase intensity. The PNF rate was significantly suppressed in the bcl-2 gene-transfer group, and tended to be reduced in the HSP70 group. CONCLUSIONS: The HVJ-liposome method effectively induced the foreign gene for kidney grafts even in the cold preservation solution. Induction of bcl-2 or the HSP70 gene reduced the occurrence of PNF in the rat renal graft. The results suggest that gene transfer not only maintains graft viability, but also graft activation. PMID- 12727471 TI - The relation between apoptosis of acinar cells and nitric oxide during acute rejection of pancreas transplantation in rats. AB - Apoptosis is an important mechanism of immune-mediated graft damage. Nitric oxide (NO) generated by inducible NO synthase (iNOS) has been demonstrated to induce apoptosis. This study investigated whether apoptosis occurs during pancreas allograft rejection and examined the relationship of apoptosis of acinar cells and NO. The rats were divided into three groups: untreated isograft group, untreated allograft group and aminoguanidine (AG)-treated group. The pancreatic grafts were harvested on the post-transplantation day 3, 5 and 7 and were used to detect the histopathological rejection grade, the expression of iNOS and the apoptotic index (AI) of the graft. iNOS presented faint positive in the acinar cells of untreated isografts and did not change greatly after transplantation (P>0.05), the level of iNOS in the untreated allografts increased progressively (P<0.01) and at the same time point was significantly higher than that of untreated isograft group and AG-treated group (P<0.01). The transferase-mediated dUTP nick end labeling showed that the apoptotic cells were mainly acinar cells. A significant correlation between AI and iNOS was noted (P<0.01, r=0.611). Therefore, NO-mediated apoptosis of acinar cells plays an important role in acute rejection of pancreas transplantation, AG can mitigate the damage of pancreas allografts. PMID- 12727472 TI - Mechanism of portal venous tolerant long-term MHC Class I L(d)-specific skin graft survival in transgenic 2CF1 mice. AB - BACKGROUND: Administration of alloantigen via the portal vein (PV) in non transgenic animals has been shown to promote immunologic tolerance and enhance transplant allograft survival. The underlying mechanisms remain unclear. In 2C x dm2 F1 (2CF1) transgenic mice, the monoclonal antibody, 1B2, identifies specific 2C TCR transgenic CD8+ T cells that are cytotoxic against Class I MHC L(d). In these mice, the specific response by these cells to L(d+) skin grafts after PV administration of L(d+) antigen was determined. MATERIALS AND METHODS: Saline (control) or allogeneic C57BL/6 x BALB/c F1 (CB6F1) spleen cells (25 x 10(6)), which differ from 2CF1 only at L(d), were injected PV into 2CF1 mice. One week later, CB6F1 tail skin was transplanted onto the dorsum of these 2CF1 mice. Skin graft rejection was defined as >50% loss of the graft. Parallel experiments were performed in non-transgenic littermates [B6F1 (C57BL/6 x dm2)]. FACS analysis of 2CF1 peripheral blood for 1B2+, CD4+, and CD8+ T cells was performed 2 days before PV injection (9 days prior to skin grafting), 5 days after PV injection (2 days prior to skin grafting), and 7, 14, 21, 28, and 60 days after skin grafting. FACS analysis of nai;ve, saline control, and CB6F1 PV-treated 2CF1 thymocytes was also performed. Responsiveness of saline (control)-treated and PV-treated 2CF1 splenocytes was measured by in vitro cytotoxic T lymphocyte (CTL). RESULTS: All CB6F1 skin grafts were rejected in <14 days by PV saline controls. However, a single PV injection of donor L(d+) CB6F1 cells was sufficient to induce indefinite CB6F1 (L(d+)) skin allograft survival in 100% of non-transgenic B6F1 and transgenic 2CF1 (anti-L(d)) TCR transgenic recipients. FACS analysis of 1B2+ T cells demonstrated that PV injection of donor antigen followed by a CB6F1 skin graft led to a 70% decrease in peripheral donor-reactive 1B2+ CD8+ T cells by day 7, while central thymocytes were unchanged. CTL of 2CF1 splenocytes following PV CB6F1 demonstrated that they were hyporesponsive to L(d) compared to saline treated 2CF1 splenocytes. Despite recovery of peripheral CD8+ T cells to near normal levels by 60 days post-transplantation, skin graft survival persisted indefinitely. CONCLUSIONS: Administration of specific PV antigen results in exquisite long-term L(d+) skin allograft acceptance. This tolerance induction is related to a significant peripheral deletion of donor-reactive 1B2+ CD8+ transgenic T cells and anergy of the residual T cells. PMID- 12727473 TI - Humoral immune responses during acute rejection in rat lung transplantation. AB - The detailed responses of humoral immunity during acute rejection remain obscure in lung transplantation (LTx). In order to clarify the reactions of alloantibodies (allo-Abs) during acute rejection, we demonstrated the time-course of changes in anti-donor Ab reaction in the peripheral blood and deposition in the grafts using a rat LTx model. Lewis (LEW) rats served as recipients for Brown Norway (BN) lung allografts (MHC fully incompatible combination). The left lung was transplanted orthotopically using a cuff technique. Syngeneic transplants (LEW to LEW) served as control. No immunosuppression therapy was administered in this model. We evaluated the alloreactivity against donor in rat recipients by detecting allo-Abs with a flow cytometric cross-match (FCXM) technique. Recipient serum samples were incubated with donor lymphocytes and stained with anti-rat immunoglobulin (Ig), to determine the titers of circulating allo-Abs in the peripheral blood with a three-color FCXM technique. We also examined the deposition of anti-donor Abs (IgG and IgM) in the grafts with an immunofluorescent method. All allografts were completely rejected and lost their aeration within 6 days after LTx. Strong allo-Abs responses of both IgG and IgM were observed in the peripheral blood during acute rejection. The level of IgM allo-Abs had already significantly increased on day 2 at the time of mild rejection; however, IgG Abs did not elevate until day 6, when the grade of rejection was severe. Circulating IgM levels started decreasing on day 8, whereas IgG Abs continued elevating. On the other hand, no evident deposition of allo-Abs in the grafts was observed until day 6. We have shown in this study that circulating IgM allo-Abs was detected at the time of mild allograft rejection, interestingly, before evident deposition in the graft. It might be suggested that allograft rejection progressed without antibody deposition until severe rejection. PMID- 12727474 TI - Hepatic expression of IL-15 mRNA is associated with liver graft acceptance. AB - BACKGROUND: Acute allograft rejection continues to be a major cause of morbidity following organ transplantation. The aim of this study was to investigate the local expression of a range of immunomodulatory molecules which may be mediating rejection of, or tolerance to, liver allografts. METHODS: RNA was extracted from 31 protocol liver biopsies taken 7-10 days post-transplantation, reverse transcribed and screened by a sensitive RT-PCR for a wide range of cytokines and other immunomodulatory molecules. The mRNA profile of each biopsy was subsequently related to the histological and clinical status of the graft. Samples of RNA isolated from activated leukocytes and T cell clones, and from normal liver, were used as controls to compare to the 'immunological snapshot' obtained from the biopsies. RESULTS: Presence of tumour necrosis factor-alpha, fas ligand, granzyme B and perforin mRNA in most of the liver biopsies reflected the occurrence of cell-mediated immune reactions. However, the expression of only one cytokine, interleukin-15 (IL-15), was significantly more frequent in allografts that showed no histological or biochemical signs of rejection during the early post-transplant period. Using an in vitro model it was demonstrated that recombinant IL-15 expands tenfold the number of CD3(+)CD56(+) (natural T; NT) cells from peripheral blood mononuclear cells cultures. Conditioning with IL 15 also increased cytotoxic activity of lymphocytes against leukaemic target cells. CONCLUSIONS: Although considerable evidence for cell-mediated immunity was shown for all liver allografts, the only clinical association was for IL-15 mRNA expression and graft acceptance. An in vitro model suggested that IL-15 may be enhancing the numbers and the activity of local regulatory cells, in particular resident NT cells in the liver, which may have a role in killing activated lymphocytes such as graft-reactive host T cells. PMID- 12727475 TI - Oncogene expression on the syngeneic beta-cells of long-term surviving pancreatic grafts and better effects of interleukin-1 receptor (IL-1R) and IL-2Rbeta on the grafted beta-cells in LEW/Sea strain rats. AB - Thirty-two normal LEW/Sea rats were transplanted a piece of syngeneic pancreas between the peritoneum and abdominal muscle. Among them, 17 (68%) of the 25 rats that received pancreatic transplantation at 41-50 days of age had a surviving beta-cell mass at 5.5-7.1 months after transplantation. Among the 25 rats, 12 rats injected with interleukin-1 receptor (IL-1R) and IL-2Rbeta peptides at post transplantation showed better surviving grafts at 5.5 months' observation. Only 2 (25%) of the other 7 young rats that received a pancreatic graft at 20 days of age had a small mass at 21 days post-transplantation. Flow cytometer (FCM) analyses showed that thymus OX40(+) (CD134(+)) T-cells were increased up to 37+/ 4% at the graft rejection in the 13 old rats without the IL-R peptide injections. The 7 young rats had 99% of thymus OX40(+) T-cells. However, the 12 old rats injected with the IL-R peptides showed suppressed numbers of thymus OX40(+) T cells (8-13+/-3%). The long-term surviving, but apoptotic, grafted beta-cells were stained positively both with anti-insulin monoclonal antibody (mAb) and with anti-c-erbB-2/human epidermal growth factor receptor (HER)-2/neu mAb. Expression of a c-erb family oncogene was shown on the pancreatic graft surviving for 7.1 months. Electron microscopic analysis of the grafted beta-cells showed abnormally large beta granules and loss of functioning mitochondria in the cytoplasm. In 18 (56%) of the 32 rats, the 220-bp and 380-bp specific products of insulin degrading enzyme (IDE) gene were amplified using the polymerase chain reaction (PCR) of the liver DNA. Among the 18 rats, 6 rats expressed 2 extra hands of 280 bp and 700-bp in a correlation with the high levels of the transforming growth factor-alpha (TGF-alpha) cDNA of 120-bp which was amplified in the quantitative reverse-transcriptase (RT)-PCR of the liver cDNA. Among the selected 11 rats, 5 rats showed large amounts of the 120-bp TGF-alpha cDNA. Host pancreatic RT-PCR showed 235-bp or 250-bp bcl-2 and 181-bp bcl-xS gene products. The bcl-2 cDNA of the host pancreas was amplified actively when the pancreatic graft was being rejected. Exceptionally, the one female injected with the IL-R peptides showed a low level of the liver TGF-alpha cDNA together with the pancreatic expressions of Bax (140-bp), bcl-2 and like interleukin converting enzyme (LICE) (318-bp) cDNA. Insulin secretion from the grafted beta-cells and IL-1beta-induced Fas-mediated apoptosis of the beta-cells were suspected to be present at the same time in the female with the best graft survival. PMID- 12727476 TI - Permanent acceptance of both cardiac and skin allografts using a mild conditioning regimen for the induction of stable mixed chimerism in mice. AB - Patients who are receiving an organ transplant nowadays are sentenced to the life long administration of immunosuppressive drugs, which have serious side effects. The reliable induction of donor-specific tolerance therefore remains a major goal in organ transplantation. Previously, we have developed a sublethal, non myeloablative murine model in which permanent mixed, multilineage chimerism and donor-specific tolerance are established. Our model involves engraftment of fully allogeneic T cell depleted donor bone marrow cells in low dose irradiated and anti-CD3 treated major histocompatibility complex (MHC)-disparate recipient mice. To investigate whether vascularized organ grafts are accepted in our model, we performed heterotopic heart transplantations in our mixed chimeric mice. Chimeric mice permanently accepted hearts from the bone marrow donor (>130 days) and rapidly rejected third party-type allografts (median survival time 9 days). Untreated control recipient mice rejected both donor- and third party-type allografts. In addition, mice that accepted their cardiac grafts, donor-specific acceptance of skin grafts was observed. In conclusion, the establishment of stable mixed chimerism with this low-toxicity regimen resulted in permanent donor specific acceptance of vascularized organ as well as skin grafts across a full MHC barrier. PMID- 12727478 TI - Time-course changes of serum immunoglobulins (IgA, IgG, IgM) after liver transplantation for alcoholic cirrhosis. AB - BACKGROUND: Serum immunoglubulin increase is a hallmark of liver disease. Serum IgA is specifically increased in alcoholic liver disease, which has been considered an IgA-associated disorder. No previous studies have been focused on the time-course changes of serum immunoglobulins (IgG, IgA, IgM) after liver transplantation. AIM OF THE STUDY: To investigate the outcome of serum immunoglobulin levels in alcoholic cirrhosis after liver transplantation, with special focus on IgA values. PATIENTS AND METHODS: A total of 18 patients, liver transplanted in our center because of alcoholic cirrhosis were included in the study. Serum immunoglobulins were assayed by nephelometry before transplantation and at different intervals after the procedure from the intraoperative period to more than a year after transplantation. RESULTS: A rapid drop in IgA, IgG and IgM concentration was observed during the surgical procedure, particularly after donor liver reperfusion, and during the first days after transplantation. Mild transient hypogammaglobulinemia (IgG) was present. After a subsequent moderate re increase, serum immunoglobulins (particularly IgA and IgG) remained stable within normal or near-normal limits during the following months after liver transplantation. CONCLUSIONS: Liver transplantation for alcoholic cirrhosis is followed by a decrease in serum IgA, IgG and IgM. In the short term, low IgG levels may be observed. In the long term, serum levels of IgA and IgG become normal or near-normal. PMID- 12727477 TI - Comparative evaluation of 'TaqMan' RT-PCR and RT-PCR ELISA for immunological monitoring of renal transplant recipients. AB - By sequentially monitoring cytokine gene expression (using RT-PCR ELISA technology) in peripheral blood cells of renal transplant recipients in the early post-operatively period we have shown that expression patterns correlate with clinical events, namely acute allograft rejection. This strategy may have the potential of predicting acute rejection prior to clinical detection. Unfortunately, the technique used was time consuming and only semi-quantitative and, therefore, not suitable for clinical application. In this study, we have sought to confirm the results of the early work using a real time quantitative RT PCR technique ('TaqMan'), which may be applicable in the clinical laboratory. 'TaqMan' primers and probes were designed for Interleukin (IL)-4 and IL-10 using Primer Express software. Cytokine gene expression for both cytokines was re measured in stored cDNA samples from 27 non-rejectors and 14 patients experiencing an episode of biopsy proven acute rejection. Compared to pre transplant levels, IL-4 gene expression fell significantly on post-operative days 2 and 7 before returning to baseline values by day 14 in the non-rejectors. In the rejectors, the initial significant fall was again seen, but with an earlier return to pre-transplant levels at the time of rejection diagnosis. This was followed by a further significant fall in levels 48 h after the initiation of anti-rejection therapy. These different patterns for rejectors and non-rejectors were seen using both techniques. For IL-10, gene expression increased significantly following transplantation throughout the study period when compared to baseline values. This pattern was seen using both techniques. In the rejectors, there were different patterns seen depending on the technique used. When using RT-PCR ELISA, the initial rise was again seen followed by a return to baseline values at the time of rejection diagnosis followed by a further significant rise in gene expression after the start of anti-rejection treatment. The pattern resembled those of the non-rejectors when expression was measured using 'TaqMan'. This study has confirmed that sequential monitoring of cytokine gene expression, measured in peripheral blood mononuclear cells, detects significant changes that correlate with clinical events in renal transplant recipients, including acute rejection, although not all changes detected with RT PCR ELISA were confirmed. Therefore, real time quantitative RT-PCR technology may be useful in monitoring the immunological status of these patients in the early post-operative period. PMID- 12727479 TI - Are corneal cells susceptible to antibody-mediated killing in corneal allograft rejection? AB - PURPOSE: The precise role of antibodies in corneal transplantation is controversial. Clinical and experimental evidence both supports and refutes the contribution of donor-derived alloantibody in corneal allograft rejection. Accordingly, we prospectively evaluated the presence of donor-derived alloantibody in two high-risk donor-host combinations. We also evaluated the ability of this alloantibody to kill corneal epithelial, keratocytes, and endothelial cells in complement-dependent and complement-independent fashions. METHODS: C3H/Hej (H-2(k)) and Balb/c (H-2(d)) corneal grafts were transplanted orthotopically to CB6F1 (H-2(b/d)) and C57BL/6 (H-2(b)) recipients, respectively. These two donor-host combinations represent disparity at the entire MHC and multiple minor histocompatibility loci. Four objectives were addressed. First, we wished to determine if there was a correlation between the appearance of donor specific serum IgG antibody and corneal graft rejection. Second, we evaluated the effect of passive transfer of hyperimmune donor-specific antibody on corneal allograft rejection. Third, we examined the capacity of donor-specific alloantibody to mediate complement-dependent cytolysis of corneal cells. Finally, we determined the capability of donor-specific alloantibody to mediate apoptosis of corneal cells. RESULTS: The presence of donor-specific serum IgG alloantibodies did not correlate with corneal graft rejection. One hundred percent of CB6F1 and C57BL/6 hosts rejected their C3H and Balb/c orthotopic corneal allografts, respectively. However, two of these seven CB6F1 hosts and one C57BL/6 host did not produce donor-specific IgG alloantibody that was significantly different from naive donors. Passive transfer of hyperimmune allo antiserum prior to corneal transplantation did not increase the incidence, severity, or tempo of corneal allograft rejection in either donor-host combination. Hyperimmune allo-antiserum produced complement-mediated lysis of C3H corneal endothelial but not C3H corneal epithelial cells in the C3H-CB6F1 donor host combination. Interestingly, all three corneal cell layers were vulnerable to complement-mediated cytolysis in the Balb/c-C57BL/6 donor-host combination. Additionally, Balb/c corneal epithelial, keratocytes, and endothelial cells were vulnerable to complement-independent, antibody induced apoptosis. CONCLUSIONS: Corneal graft rejection does not appear to correlate with the production of IgG alloantibody and can occur in the absence of donor-specific IgG alloantibody. Antibody-mediated killing of the corneal endothelium can occur in a complement dependent or complement-independent fashion. PMID- 12727480 TI - Ex vivo and systemic transfer of adenovirus-mediated CTLA4Ig gene combined with a short course of FK506 therapy prolongs islet graft survival. AB - Adenovirus-mediated CTLA4Ig gene transfer has been reported to enhance graft survival in several rodent transplantation models. In this study, we investigated the efficacy of ex vivo and systemic transfer of the CTLA4Ig gene by adenoviral vectors in pancreatic islet allo-transplantation. Islet grafts from BN rats were transplanted to chemically induced diabetic LEW rats. First, ex vivo CTLA4Ig gene transfer into isolated islets was performed prior to transplantation. Survival of transduced grafts under the kidney capsule was slightly prolonged (8.6+/-1.3 days) compared with survival of untransduced grafts (6.7+/-1.2 days); when combined with a short course of FK506, graft survival was further extended (32.6+/-10.7 days vs. 13.7+/-1.0 days with FK506 alone). Secondly, systemic gene transfer was accomplished by intravenous administration immediately after the transplantation procedure. In these animals, islet grafts under the kidney capsule survived longer (15.2+/-3.3 days) than in controls (6.7+/-1.2 days), and when FK506 was administered perioperatively, all the islet grafts survived for more than 100 days. In systemically transduced recipients, the survival of islet grafts transplanted into the liver was not significantly different from that of the grafts placed under the kidney capsule. In order to examine organ-specific immunogenicity, heterotopic BN cardiac grafts were transplanted to LEW rats intra abdominally, with the virus transferred systemically as in the islet model. In contrast to the islet grafts, all the cardiac grafts were accepted for longer than 100 days, even without FK506 therapy. Finally, the LEW recipients with long surviving islet or cardiac grafts were re-transplanted with islet grafts from the same donor strain (BN) on day 100. The second islet grafts survived longer than 100 days in half of the cardiac recipients, but consistently failed in the islet recipients. We conclude that in this transplant model, CTLA4Ig gene transfer and FK506 treatment synergistically improved islet graft survival, systemic transfer of the gene was more effective than ex vivo transfer to the islets, and donor specific tolerance could not be achieved for islet transplantation but was achieved for cardiac transplantation. PMID- 12727481 TI - Increased IFN-gamma response to transplantation antigens measured by cytokine MLC: indications for a bi-phasic response pattern. AB - The immunological reactivity to major histocompatibility transplantation antigens (MHC) is important in clinical manifestations such as graft rejection and graft vs.-host disease. To evaluate the allogeneic cytokine response pattern we used a newly developed cytokine mixed leukocyte culture (MLC) technique. Blood mononuclear cells from healthy women (n=6) were exposed to cells from another person and to pooled cells from 28 blood donors (MHC-pool). The secretion kinetics of IL-4 and IFN-gamma from the responder cells was analysed by ELISPOT. We found a higher IFN-gamma response to the MHC-pool compared with the IL-4 secretion. Both the total secretion of IFN-gamma for 7 days and the median value of IFN-gamma in each individual was increased compared with IL-4. The IFN-gamma response showed a bi-phasic pattern with the major peak on day 6-7. Our results indicate that allo-responses are mainly Th1-like responses, displaying a bi phasic pattern. This knowledge may be useful, and the methods suitable, in the studies of allo-responses in transplantation and pregnancy. PMID- 12727482 TI - Combined analysis of cytokine genotype polymorphism and the level of expression with allograft function in African-American renal transplant patients. AB - Cytokine gene polymorphism and expression levels were evaluated in a group of African-American patients who had undergone renal transplantation. It was hypothesized that possession of specific cytokine alleles might be influential in predisposing the recipient to allograft rejection. Thus, we sought to establish a relationship between cytokine gene polymorphism, the levels of cytokine expression, and the outcome of allograft function. Cytokine genotypes and mRNA transcript levels of IL-2, TNF-alpha, TGF-beta1, IL-10, IL-6 and IFN-gamma were determined using peripheral blood cells. Genomic DNA samples from 77 transplant recipients and 77 controls were tested by a multiplex PCR with specific primers for the above cytokines. The frequency distributions of cytokines were analyzed in respect to the clinical characterization, including delayed graft function (DGF), rejection episodes (REs) and stable graft function (SGF). The mRNA transcript level was tested both at pre- and early post-transplantation (day 1 and day 4) with primers for coding regions of the above cytokines in a RT-PCR assay. The majority of recipients with successful graft function were matched with their donors for only three out of the six HLA alleles. We have shown that the TGF-beta1 T/C G/G high producer and IFN-gamma T/A intermediate producer genotypes were associated with allograft rejection, whereas low IFN-gamma producer and high IL-10 producer genotypes were significantly protective of the allograft. There was some correlation between the TGF-beta1 high producer genotype and DGF, but it was not statistically significant. Overall, 77% of those who experienced REs carried the TGF-beta1 T/C G/G, high producer genotype as compared with 52% who experienced DGF, 39% with SGF (P<0.01, RR=2.0), and 27.3% of controls (P<0.003, RR=2.6). The IFN-gamma T/A intermediate producer genotype was found in 69.2% of patients with REs as compared with 26.8% of patients with SGF (P<0.008, RR=2.85). The IL-10, ATA/ATA low producer genotype was found in 38.5% of recipients with REs and 14.6% of recipients without REs (P<0.04, RR=0.53). Expression levels of mRNA transcript were correlated with genotype data, except for the TGF-beta1 high producer genotype where there was no significant difference between the level of mRNA transcript at pre- and post transplantation. Low DRbeta1 and high DPbeta1 expression by recipient peripheral blood mononuclear cells before transplantation was associated with more SGF, whereas high DRbeta1 and low DPbeta1 expression at pretransplantation was associated with more REs (DRbeta1, P<0.001 and DPbeta1, P<0.05, respectively). We concluded that, dual analysis of cytokine genotype and expression levels by peripheral cells may be an important clue to understanding the contribution of the recipient's immune response to an allograft pre- and post-transplantation. Identification of peripheral markers diagnostic of rejection could allow advance anticipation of clinical outcome, and might reduce the need for tissue biopsy. PMID- 12727483 TI - Recipient gene polymorphisms in the Th-1 cytokines IL-2 and IFN-gamma in relation to acute rejection and graft vascular disease after clinical heart transplantation. AB - IL-2 and IFN-gamma are associated with acute rejection (AR) and graft vascular disease (GVD) after clinical heart transplantation. Polymorphisms in the genes of IL-2 (T-330G in the promoter) and IFN-gamma (CA repeat in the first intron) influence the production levels of these cytokines. Therefore, these polymorphisms might have an effect on the outcome after transplantation. To investigate possible effects of genetic variations in IL-2 and IFN-gamma genes on AR and GVD, we analyzed the IL-2 T-330G and the IFN-gamma CA repeat polymorphism in DNA of 301 heart transplant recipients. No associations were found for allele or genotype distributions between patients with or without AR (IL-2 allele frequency: P=0.44, genotype distribution: P=0.46; IFN-gamma allele frequency P=0.10, genotype distribution 12 repeats allele: P=0.21). Also, no associations were found analyzing the number (0 vs. 1 vs. >or=1) of AR (IL-2 allele frequency: P=0.59; genotype distribution: P=0.37; IFN-gamma allele frequency: P=0.27, genotype distribution 12 repeats allele: P=0.41) or analyzing the polymorphisms in patients with AR within the first month or thereafter (IL-2 allele frequency: P=0.45, genotype distribution: P=0.38; IFN-gamma allele frequency: P=0.21, genotype distribution 12 repeats allele: P=0.41). Analyzing both polymorphisms in relation to GVD, resulted in comparable allele and genotype distributions (IL-2 allele frequency: P=0.75; genotype distribution: P=0.77; IFN-gamma allele frequency: P=0.70, genotype distribution 12 repeats allele: P=0.63). In conclusion, we did not detect an association between the IL-2 T-330G promoter polymorphism and CA repeat polymorphism in the first intron of the IFN-gamma gene and AR or GVD after heart transplantation. PMID- 12727484 TI - Morpho-histological studies in the aromatic species of Chenopodium from Argentina. AB - A morpho-histological study of the vegetative organs (stem and leaf) of the aromatic species of Chenopodium L. from Argentina [C. ambrosioides L., C. burkartii (Aellen) Vorosch., C. carinatum R. Br., C. chilense Schrad., C. graveolens Willd. var. bangii (Murr) Aellen, C. haumanii Ulbr., C. multifidum L., C. oblanceolatum (Speg.) Giusti, C. pumilio R. Br., C. retusum (Moq.) Moq., and C. venturii (Aellen) Cabrera] was carried out. Classifications for the glandular and non-glandular trichomes are established and their presence among species is presented. A variant in both the dorsiventral and isobilateral mesophyll is reported; some data are valuable for systematic purposes and for the identification of dried and smashed material used as vegetal drug. PMID- 12727485 TI - Antimycobacterial flavones from Haplopappus sonorensis. AB - Crude extracts of Haplopappus sonorensis (A. Gray) S.F. Blake (Asteraceae), showed activity against Mycobacterium tuberculosis H(37)Rv. By assay-guided fractionation, 5-hydroxy-3,7,4'-trimethoxyflavone (1). 5,7-dihydroxy-3,4' dimethoxyflavone (2). and 5,4'-dihydroxy-3,7-dimethoxyflavone (3). were identified as the antimycobacterial principles. Compound 2 was the most active compound. PMID- 12727486 TI - Analgesic, antipyretic and anti-inflammatory effects of essential oil of Lippia multiflora. AB - The essential oil of Lippia multiflora Moldenke (Verbenaceae) produced by conventional hydrodistillation was analyzed and studied for its analgesic, antipyretic and anti-inflammatory activities in rats and mice. At the doses used (2, 4 and 8 ml/kg o.s.) the essential oil of L. multiflora showed significant and dose-dependent analgesic effect on acetic acid-induced writhing in mice. Only the dose of 8 ml/kg of essential oil, antagonized hyperexia induced by brewer's yeast. No effect on granuloma formation was observed. PMID- 12727487 TI - Triterpenoid saponins from the roots of Silene cucubalus. AB - A new triterpenoid saponin was isolated from the roots of Silene cucubalus and its structure was determined on the basis of one- and two-dimensional NMR spectroscopy as gypsogenin 3-O-beta-xylopyranosyl-(1-->3)-[beta-galactopyranosyl (1-->2)]-beta-glucuronopyranoside. A known saponin, quillaic acid 3-O-beta xylopyranosyl-(1-->3)-[beta-galactopyranosyl-(1-->2)]-beta-glucuronopyranoside, was also isolated. PMID- 12727488 TI - A comparative study of Ferula hermonis root extracts and sildenafil on copulatory behaviour of male rats. AB - The effect of 600 mg/kg given by oral route to rats of Ferula hermonis roots extracts (petroleum ether, ethyl acetate, methanol and water) was evaluated on sexual behaviour of male rats. Petroleum ether and ethyl acetate extracts produced a significant decrease in both the mount rate (MR) and the intromission rate (IR), significant prolongation of intromission latency (IL) was observed when these extracts were compared with both controls and sildenafil. Methanolic extract produced a significant increase in MR while no effect has been observed on IR or IL in comparison with control. The effect of water extract was not significantly different from controls for the MR and IR, but there was a significant prolongation in the IL. PMID- 12727489 TI - A simple HPLC-UV method for the assay of ginkgolic acids in Ginkgo biloba extracts. AB - Extracts from the leaves of Ginkgo biloba are among the most widely used phytotherapeutics. Some alkylphenols (ginkgolic acids, cardanols and cardols) have been described as potentially hazardous constituents in Ginkgo extracts. Accordingly, a requirement for a maximum concentration of ginkgolic acids has been proposed in the UE and US pharmacopoeias Ginkgo monographs establishing a limit value of 5 ppm. A novel HPLC-UV method, developed by the use of HPLC-APCI MS HPLC-DAD techniques and allowing the identification of ginkgolic acids and related phenols, is described. The new analytical method, not requiring enrichment procedures, can be used for the quantification of ginkgolic acids in Ginkgo extracts. PMID- 12727490 TI - Studies on the immunomodulatory activity of flavonoidal fraction of Tephrosia purpurea. AB - The flavonoid fraction of Tephrosia purpurea (FFTP) was studied for its effect on cellular and humoral functions and on macrophage phagocytosis in mice. Oral administration of FFTP (10-40 mg/kg) significantly inhibited sheep red blood cells (SRBC)-induced delayed-type hypersensitivity reactions. It also produced a significant, dose-related decrease in sheep erythrocyte-specific haemagglutination antibody titre. However, the fraction failed to show a significant change in the macrophage phagocytic activity. The results obtained indicate the ability of the flavonoidal fraction of T. purpurea to modulate both the cell-mediated and the humoral components of the immune system. PMID- 12727491 TI - Studies on analgesic activity of Cleome viscosa in mice. AB - The analgesic activity of methanol extract of Cleome viscosa, given orally at the doses of 100, 200, 400 mg/kg was evaluated for its analgesic activity in mice using the acetic acid-induced writhing and the tail flick, tail clip, tail immersion methods. The extract showed promising activity in all the tests. PMID- 12727492 TI - Effect of Garcinia cambogia extract on serum leptin and insulin in mice. AB - In this study we examined the effects of 3.3% Garcinia cambogia extract on 10% sucrose loading in mice for 4 weeks. Treatment was found to have no effect on body weight, fat pad weight or serum glucose level. On the other hand, serum total cholesterol, triglycerides, NEFA were observed. Levels of serum insulin and leptin, as well as the leptin/WAT ratio, were lower in the treated mice than in the control. These findings suggested that G. cambogia extract efficiently improved glucose metabolism and displayed leptin-like activity. PMID- 12727493 TI - Antihyperglycemic activity of Caralluma attenuata. AB - Extracts of Caralluma attenuata are screened for their antihyperglycemic activity. Ethanol, chloroform and butanol extracts were tested on glucose loaded and alloxan-induced diabetic rats. In both tests, the butanol extract, at the oral dose of 250 mg/kg, has shown statistically significant and considerable antihyperglycemic activity. PMID- 12727494 TI - Effect of Indigofera aspalathoides against Dalton's ascitic lymphoma. AB - The effect of intraperitoneal injection of 400 mg/kg of ethanolic extract of Indigofera aspalathoides (EEIA) against Dalton's ascitic lymphoma has been evaluated in Swiss albino mice. A significant increase in the life span, a decrease in the cancer cell count and body weight were noted in the tumour induced mice after treatment with EEIA. These observations indicate that the plant is having protective effect in Dalton's ascitic lymphoma. PMID- 12727495 TI - Antimicrobial study of essential oils of Ocimum gratissimum leaves and Zanthoxylum xanthoxyloides fruits from Cameroon. AB - An investigation of antimicrobial activities of essential oils of fresh leaves of Ocimum gratissimum and the essential oil of the dried fruits of Zanthoxylum xanthoxyloides was carried out. The essential oils showed extensive inhibition zones and are, therefore, effective antimicrobial systems. PMID- 12727496 TI - Antifungal activity of a thiophene polyine from Leuzea carthamoides. AB - The thiophene polyine (E)-2-[5-(hept-5-en-1,3-diynyl)-thien-2-yl]-ethan-1,2-diol was isolated from an ethanolic extract of underground parts of Leuzea carthamoides. The thiophene polyine demonstrated significant antifungal activity against all tested species. PMID- 12727497 TI - Antimicrobial activity of Ixora coccinea leaves. AB - Ether and methanol extracts of Ixora coccinea dry leaves were tested for their antimicrobial activity. Ether fraction was found to be more active than the methanol fraction. PMID- 12727498 TI - Antifungal activity of Terminalia australis. AB - Dichloromethane, methanol and aqueous extracts of the aerial parts of Terminalia australis were evaluated for their antifungal activity. The methanol and aqueous extracts were found to be effective against the tested Aspergillus and Candida strains. PMID- 12727499 TI - Molluscicidal and antimicrobial activity of Solanum aculeastrum. AB - The methanolic extract of the fresh root bark and berries of Solanum aculeastrum showed significant activity against host snails of schistosomiasis. The berries extract was more potent with 100% snail kill at 50 ppm. Fractionation increased activity with 100% mortality at 20 ppm. The aqueous and methanolic extracts of the berries showed moderate antimicrobial activity, increasing by fractionation. PMID- 12727500 TI - Preliminary evaluation of inhibition of matrix-metalloprotease MMP-2 and MMP-9 by Passiflora edulis and P foetida aqueous extracts. AB - Fruit's decoctions of Passiflora edulis and P. foetida var. albiflora were evaluated for the inhibition of activity of gelatinase MMP-2 and MMP-9, two metallo-proteases involved in the tumour invasion, metastasis and angiogenesis. Both water extracts, at different concentrations, inhibited the enzymes. PMID- 12727501 TI - Biological activity of Euonymus europaeus. AB - The n-hexane, dichloromethane and methanol extracts of the seeds of Euonymus europaeus have been screened for antibacterial and free radical scavenging activity. General toxicity (brine shrimp lethality assay) of these extracts has also been assessed. PMID- 12727502 TI - Antimicrobial and cytotoxic constituents of Loranthus globosus. AB - (+)-Catechin, 3,4-dimethoxycinnamyl alcohol and 3,4,5-trimethoxycinnamyl alcohol were isolated from the barks of Loranthus globosus. All compounds showed significant antimicrobial and cytotoxic activities. PMID- 12727503 TI - In vitro activities of Maesa lanceolata extracts against fungal plant pathogens. AB - In vitro tests were carried out using extracts of Maesa lanceolata var. goulungensis weir against a broad range of fungal plant pathogens such as Phytophthora cryptogea, Trichoderma virens, Aspergillus niger, Phoma sp., Fusarium oxysporium, Pythium ultimum, Cochliobolus heterostrophus, Rhizoctonia solani, Sclerotium rolfsii and Pyrenophora teres. M. lanceolata extracts were very active against all the pathogens tested except P. ultimum and R. solani. PMID- 12727504 TI - Hepatoprotective effect of Foeniculum vulgare essential oil. AB - Hepatoprotective activity of Foeniculum vulgare (fennel) essential oil (FEO) was studied using carbon tetrachloride (CCl(4)) induced liver injury model in rats. The hepatotoxicity produced by acute CCl(4) administration was found to be inhibited by FEO with evidence of decreased levels of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and bilirubin. The results of this study indicate that FEO has a potent hepatoprotective action against CCl(4)-induced hepatic damage in rats. PMID- 12727505 TI - A new triterpenoid from Lantana camara. AB - A new ursane was isolated from the leaves of Lantana camara and its structure elucidated as 3,24-dioxo-urs-12-en-28-oic acid (1) by means of spectral analysis. PMID- 12727507 TI - Emerging ideas on the molecular basis of protein and peptide aggregation. AB - Several neurodegenerative diseases are associated with the unfolding and subsequent fibrillization of proteins. Although neither the assembly mechanism nor the atomic structures of the amyloid fibrils are known, recent experimental and computational studies suggest that a few general principles that govern protein aggregation may exist. Analysis of the results of several important recent studies has led to a set of tentative ideas concerning the oligomerization of proteins and peptides. General rules have been described that may be useful in predicting regions of known proteins (prions and transthyretin) that are susceptible to fluctuations, which give rise to structures that can aggregate by the nucleation-growth mechanism. Despite large variations in the sequence dependent polymerization kinetics of several structurally unrelated proteins, there appear to be only a few plausible scenarios for protein and peptide aggregation. PMID- 12727508 TI - Minimalist models for protein folding and design. AB - Protein folding research during the past decade has emphasized the dominant role of native state topology in determining the speed and mechanism of folding for small proteins; this has been illustrated by simulations using minimalist protein models. The advantages of minimalist protein models lie in their ability to rapidly collect meaningful statistics about folding pathways and kinetics, their ease of characterization with coarse-grained order parameters and their concentration on the essential physics of the problem to connect with experimental observables for a target protein. The maturation of experimental protein folding has driven the need for more quantitative protein simulations to better understand the balance between sequence details and fold topology. In the past year, we have seen the emergence of more complex minimalist models, ranging from all-atom Go potentials to coarse-grained bead models in which Go interactions are replaced or supplemented by more physically motivated potentials. The reduced computational cost at the coarse-grained level of abstraction will potentially enable both folding studies on a genomic scale and systematic application in protein design. PMID- 12727509 TI - Peptide folding simulations. AB - Developments in the design of small peptides that mimic proteins in complexity, recent advances in nanosecond time-resolved spectroscopy methods to study peptides and the development of modern, highly parallel simulation algorithms have come together to give us a detailed picture of peptide folding dynamics. Two newly implemented simulation techniques, parallel replica dynamics and replica exchange molecular dynamics, can now describe directly from simulations the kinetics and thermodynamics of peptide formation, respectively. Given these developments, the simulation community now has the tools to verify and validate simulation protocols and models (forcefields). PMID- 12727510 TI - New approaches to the dynamic interpretation and prediction of NMR relaxation data from proteins. AB - NMR relaxation experiments of isotopically labeled proteins provide a wealth of information on reorientational global and local dynamics on nanosecond and subnanosecond timescales for folded and nonfolded proteins in solution. Recent methodological advances in the interpretation of relaxation data have led to a better understanding of the overall tumbling behavior, the separability of internal and overall motions, and the presence of correlated dynamics between different nuclear sites, as well as to new insights into the relationship between reorientational dynamics and primary and tertiary protein structure. Some of the new methods are particularly useful when dealing with nonfolded protein states. PMID- 12727511 TI - Catalysis by enzyme conformational change as illustrated by orotidine 5' monophosphate decarboxylase. AB - An energy decomposition scheme has been used to elucidate the importance of the changes of enzyme conformational substates to the reduction of the activation barrier in enzyme-catalyzed reactions. This analysis may be illustrated by the reaction of orotidine 5'-monophosphate decarboxylase, which exhibits a remarkable rate enhancement of over 17 orders of magnitude compared to the uncatalyzed process. The mechanism shows that the enzyme conformation is more distorted in the reactant state than in the transition state. The energy released from protein conformation relaxation provides the predominant contribution to the rate enhancement of orotidine 5'-monophosphate decarboxylase. The proposed mechanism is consistent with results from site-directed mutagenesis experiments, in which mutations distant from the reactive center can have significant effects on the catalytic rate enhancement (k(cat)), but rather a small influence on the binding affinity for the substrate (K(M)). PMID- 12727512 TI - Biological networks. AB - Recent advances in high-throughput methods have provided us with a first glimpse of the overall structure of molecular interaction networks in biological systems. Ultimately, we expect that such information will change how we think about biological systems in a fundamental way. Instead of viewing the genetic parts list of an organism as a loose collection of biochemical activities, in the best case, we anticipate discrete networks of function to bridge the gap between genotype and phenotype, and to do so in a more profound way than the current qualitative classification of linked reactions into familiar pathways, such as glycolysis and the MAPK signal transduction cascades. At the present time, however, we are still far from a complete answer to the most basic question: what can we learn about biology by studying networks? Promising steps in this direction have come from such diverse approaches as mathematical analysis of global network structure, partitioning networks into functionally related modules and motifs, and even de novo design of networks. A complete picture will probably require integrating the data obtained from all of these approaches with modeling efforts at many different levels of detail. PMID- 12727514 TI - Structural insights into BRCA2 function. AB - BRCA2 is a tumor suppressor directly implicated in familial breast cancer. Extensive genetic and biochemical characterization has shown that BRCA2 is involved in the maintenance of chromosomal stability and that it has an important role in recombination-mediated double-strand DNA break repair. Two recent structures of BRCA2 domains have revealed that it may serve as a critical mediator of DNA repair through direct interactions with Rad51, the eukaryotic homolog of RecA, and with single-stranded DNA. Before the structures were determined, little was known about the structural basis of BRCA2 interactions with the Rad51 pathway of DNA repair. Taken together, the structures provide striking insights into the role of BRCA2 in double-strand DNA break repair and suggest a direct role for BRCA2 in homologous recombination that was not evident from earlier studies. PMID- 12727515 TI - The chemistry of protein synthesis and voyage through the ribosomal tunnel. AB - High-resolution structures of ribosomal subunits and their complexes with substrates and antibiotics have revealed fundamental principles of template directed protein synthesis. Mechanistic questions regarding ribosome function and catalysis can now be addressed with structure-based experiments. Recent studies have investigated the mechanism of peptide bond formation catalyzed by the large ribosomal subunit, the mode of protein synthesis inhibition by macrolide antibiotics, the interaction of nascent polypeptides with the ribosomal exit tunnel, and the role of ribosomal proteins in the recruitment of accessory factors that assist protein folding and targeting. PMID- 12727516 TI - Surprising cofactors in metalloenzymes. AB - Transition metal complexes are located at the active sites of a number of enzymes involved in intriguing biochemical reactions. These complexes can exhibit a wide variety of chemical reactivity due to the ease at which transition metals can adopt different coordination environments and oxidation states. Crystallography has been a powerful technique for examining the structure and conformational variability of complex biological metallocenters. In particular, the past ten years have provided a wealth of structural information and several surprises concerning the metallocenters at the active sites of nitrogenase, hydrogenase and carbon monoxide dehydrogenase/acetyl-coenzyme A synthase. PMID- 12727517 TI - Protein-protein unbinding induced by force: single-molecule studies. AB - Experiments in which two specifically interacting protein molecules are dissociated by external force have yielded new insights into mechanisms involved in cell adhesion, leukocyte rolling, regulation of integrin activity, antigen antibody interactions and other protein-mediated reactions contingent upon molecular recognition. Another important aspect of force-induced protein-protein unbinding studies is the new information being gleaned about the thermodynamics and kinetics of bond rupture. PMID- 12727518 TI - Mapping functional residues onto integrin crystal structures. AB - Historical data, available in the literature, concerning the sites responsible for integrin-ligand binding and activation may now be transposed onto recently solved crystal structures of integrin dimers and functional domains. The data were obtained through studies of natural and engineered mutations within integrins, and analyses of the epitopes for function-altering monoclonal antibodies. In the past two years, this combining of structure and function has led to the development of novel hypotheses for the mechanisms of integrin regulation. PMID- 12727519 TI - Actin's prokaryotic homologs. AB - Actin is one of the most abundant and conserved eukaryotic proteins. Remarkably, two prokaryotic homologs of actin, MreB and ParM, have only recently been identified. MreB and ParM polymerize into filaments and play important roles in the control of bacterial cell shape and plasmid segregation, respectively. Whereas the eukaryotic actins display a remarkable degree of conservation (e.g. no amino acid changes in muscle actin from chickens to humans), the two bacterial proteins have as much sequence similarity to each other ( approximately 11% sequence identity) as they do to actin. It is possible that the interesting properties of eukaryotic F-actin may account for the unusual degree of conservation among the actins, whereas the bacterial proteins have had fewer constraints over the course of evolution. PMID- 12727520 TI - Rad50/SMC proteins and ABC transporters: unifying concepts from high-resolution structures. AB - ATP-binding cassette (ABC)-type ATPases are chemo-mechanical engines for diverse biological pathways. ABC ATPase domains act not only in ABC transporters but also in DNA mismatch, nucleotide excision and double-strand break repair enzymes, as well as in chromosome segregation. Atomic-resolution crystal structures suggest molecular mechanisms for ABC ATPases and reveal surprisingly significant mechanistic and architectural conservation. This emerging unified structural biochemistry provides general medical and biological insights into how ABC proteins function as chemo-mechanical devices. ATP binding by the signature and Q loop motifs drives the conformations of substrate-specific domains to accomplish diverse functions in transmembrane transport and DNA repair. PMID- 12727523 TI - Dengue and dengue hemorrhagic fever in the Americas: lessons and challenges. AB - The incidence of dengue and dengue hemorrhagic fever (DF/DHF) has increased significantly over the last decades. Yearly, an estimated 50-100 million cases of DF and about 250000-500000 cases of DHF occur worldwide. The epidemiological situation in Latin America now resembles that in Southeast Asia. Here, the main clinical, epidemiological and virological observations in the American region are presented and compared with those previously reported from Southeast Asia. During 2002, more than 30 Latin American countries reported over 1000000 DF cases. DHF occurred in 20 countries with more than 17000 DHF cases, including 225 fatalities. The co-circulation of multiple serotypes has been reported from many countries. In the Americas, DHF is observed both in children and adults; secondary infection by a different dengue virus serotype has been confirmed as an important risk factor for this severe form of the disease. However, some new risk factors such as the interval of dengue virus infections and the ethnicity and underlying chronic conditions of the patient have also been identified. The sequence of dengue virus infections and association with certain genotypes are further factors of importance. We also discuss the control and prevention strategies. In conclusion, without urgent action for the prevention and control of dengue/DHF and its vector, the current situation will worsen and, more dramatical, there is a risk of the urbanization of yellow fever. PMID- 12727521 TI - Tubulin rings: which way do they curve? AB - Tubulin is known to exist in at least two main conformations: straight when bound to GTP or buried within the microtubule lattice, and curved when bound to GDP. The latter is most obvious during microtubule depolymerization, when protofilaments bend and peel off from microtubule ends. The curved, low-energy subunits form tantalizing ring structures in the presence of stabilizing divalent cations. Interestingly, cellular factors and antimitotic agents that act by depolymerizing microtubules can induce the formation of rings. In these rings, tubulin dimers generally appear kinked at the monomer-monomer interface, either to the same or to a lesser extent than at the dimer-dimer interface, with each agent giving rise to particular subtleties in the structures of the rings and the tubulin dimer itself that may reflect their distinctive mechanisms of action. How these kinks relate to what happens when the stored energy of GTP hydrolysis is released, freeing GDP*tubulin into an unconstrained state, remains an open question. PMID- 12727524 TI - Respiratory syncytial virus in healthy adults: the cost of a cold. AB - Respiratory syncytial virus (RSV) is well recognized as a major pathogen of lower respiratory tract infection and hospitalization in young infants. More recently the pathogenicity of RSV has been demonstrated in elderly adults, institutionalized individuals, and those with compromised immune function. In these populations RSV spreads with ease and frequently results in severe or fatal cardiopulmonary complications. In younger, healthy adults, however, the manifestations and importance of RSV infection have been studied little, and RSV is generally not considered as a cause of respiratory illness in this healthy, working population. RSV occurs in yearly outbreaks and is highly contagious. Immunity after infection is neither complete nor durable. Repeated infections, therefore, occur throughout life. In most cases these recurrent infections involve the upper respiratory tract and thus do not receive a specific diagnosis. However, recent studies indicate that in the younger, healthy adult these respiratory illnesses tend to be more severe than the average 'cold' and may have manifestations similar to influenza. An appreciable proportion results in work absence. Thus, the emerging information suggests that RSV infection clearly occurs frequently in healthy adults in contact with children, but is generally not diagnosed. The potential burden on the healthcare system is unestimated, possibly unappreciated, and should be considered in strategies being developed for preventing RSV infection. PMID- 12727525 TI - Emergence of herpes simplex type 1 as the main cause of recurrent genital ulcerative disease in women in Northern Ireland. AB - BACKGROUND: Genital herpes is a common infection affecting some 20% of sexually active people. Although herpes simplex virus (HSV) types 1 and 2 can both establish genital latency, reactivation from the sacral ganglia favours HSV-2. Over the past decade the incidence of type 1 genital infection in women has greatly increased. OBJECTIVES: To determine whether the increased prevalence of HSV-1 genital infection was benign or influencing the pattern of virus recovery in recurrent infection. STUDY DESIGN: A retrospective analysis of laboratory computer records was undertaken. Patients attending six genitourinary medicine (GUM) departments, over an 80 months period, were identified. Recurrent infection was confirmed where virus was recovered from at least two separate episodes of genital ulceration that were separated by an interval of 12 or more weeks. Episodes were further analysed for frequency, age, gender and virus type. RESULTS: Sixty nine patients with recurrent genital herpetic infection were identified. HSV-1 and HSV-2 were predominantly recovered from recurrent genital infections in females (34 HSV-1 vs. ten HSV-2) and males (one HSV-1 vs. 24 HSV 2), respectively (P>0.001). The mean age of females and males, at the initial diagnosis, was 26 and 39 years. There was no difference in the recurrence rate by type. CONCLUSIONS: HSV-1 has become the commonest cause of recurrent genital ulceration in Northern Ireland, almost entirely due its recent increased prevalence in women over the last decade. Women are experiencing genital herpetic infections at an earlier age than men. PMID- 12727527 TI - Definition of two new epitopes on human immunodeficiency virus type 1 gag protein recognized by human CD8+ cytotoxic T lymphocyte clones. AB - We have established 3 CD8+ cytotoxic T lymphocyte (CTL) clones from the peripheral blood mononuclear cells of two HIV-1-seropositive asymptomatic donors. The epitopes recognized by these CTL clones were defined using synthetic peptides. The epitopes were located on HIV-1gag protein between amino acid (a.a.) 145 and 155 (QAISPRTLNAW), a.a.193 and 201 (GHQAAMQML), and a.a.260 and 267 (EIYKRWII), and were presented by HLA-A25, HLA-B38 and HLA-B8, respectively. The former 2 epitopes have not been previously defined. The HLA-A25-restricted epitope overlapped with HLA-B57-restricted and HLA-Cw3-restricted epitopes previously reported. In addition, this epitope overlapped with an HLA-DQ restricted epitope recognized by CD4+ CTL. The HLA-B38-restricted epitope overlapped with HLA-A2-restricted and HLA-Bw52-restricted epitopes that were previously reported. The HLA-B38-restricted epitope between a.a.193 and 201 was highly conserved among HIV-1 strains. The results demonstrate that two new epitopes were defined in a region of gag protein that includes multiple epitopes presented by multiple HLA. PMID- 12727526 TI - Universal primers for real-time amplification of DNA from all known Orthohepadnavirus species. AB - BACKGROUND: The family of Hepadnaviridae is made up of members infecting birds (genus Avihepadnavirus) or mammals (genus Orthohepadnavirus). Hepatitis B virus (HBV), the hepadnavirus infecting humans, can be divided into the seven genotypes A-G. By definition, genotypes differ by more than 8% at the nucleotide level. However, some genotypes differ by more than 14% from others. OBJECTIVES: The diversity of HBV genotypes necessitates great care in primer design to find primers suitable for routine diagnostic procedures that are highly conserved. Our aim was to find a target sequence on the HBV genome that is highly conserved among all known orthohepadnaviruses, to avoid false-negative polymerase chain reaction (PCR) results due to uncommon variants of HBV. METHODS: Using an alignment of 177 genomes of orthohepadnaviruses from GenBank, we selected a primer pair from a highly conserved region, corresponding to hydrophobic transmembrane domains of the major surface protein of HBV. RESULTS: The primer pair chosen was suitable to amplify genome sequences from HBV and to the genetically most distant woodchuck hepatitis virus in real-time PCR using the LightCycler, Roche. Moreover, the primers were suitable for accurate quantitation of both viral genomes over a range from 100 to 10(10) genomes/ml. CONCLUSION: The described primers are useful for reliable detection and accurate quantitation of all known hepadnaviral genomes and may be used for the search for unknown orthohepadnaviruses. PMID- 12727528 TI - RSV molecular characterization and specific antibody response in young children with acute lower respiratory infection. AB - The presence of respiratory syncytial virus (RSV) in nasopharyngeal aspirates (NPA) were studied in 254 hospitalized Argentinean children with acute lower respiratory infection tract (ALRI). The specific humoral immune response and partial sequences of the G protein gene were studied in a subset of 22 children with RSV confirmed infection. The RSV IgM detection and the RSV IgG titration were made by immunofluorescence assay (IFA) in pairs of sera. The partial RSV G gene sequences were obtained by an RT-PCR amplification directly from de NPAs. RSV was present in 44.5% of the children. The RSV IgM was detected in 22.7 and 68.8% of the first and second sera, respectively. The IgG geometric mean titers of the acute and convalescent sera were 8 and 589. The RSV IgG titration was able to define 86.4% of the RSV confirmed cases. The percentage of coincidence between RSV IgM detection in the second sera and diagnosis by RSV IgG titration was 72.7% and no significant differences were observed. The nucleotide sequence of one group A and three group B viruses were identified. The first one was related with circulating viruses in Madrid, Montevideo and Mozambique during 1992, 1989 and 1999, respectively. The three sequences identified as group B viruses were closely related with circulating viruses in 1998 from South Africa and Canada during 1999 and 2000. The data obtained in our study provide the first approach at the molecular level (nucleotide) of the RSV circulating strains in Argentina and the lack of genotype patterns previously determined make necessary a continuous molecular surveillance in order to contribute to the understanding of the behavior of this virus in our community. PMID- 12727530 TI - Similar increased serum dipeptidyl peptidase IV activity in chronic hepatitis C and other viral infections. AB - BACKGROUND: Dipeptidyl peptidase IV is a transmembrane enzyme widely expressed in many cell types, but also present as a soluble form in biological fluids. Its abnormal activity is sometimes associated with liver disease related pathologies. OBJECTIVES: The aim of this study was to evaluate the clinical relevance of changes in serum DPPIV activity in hepatitis C and other viral infections. STUDY DESIGN: DPPIV activity was assessed by using a microplate-based colorimetric assay on serum from 88 subjects: 12 healthy uninfected controls, 10 patients with primary biliary cirrhosis (PBC) as a reference group, 36 HCV-infected patients, and patients suffering from viral infections of different etiologies. Levels of DPPIV activity were compared with: (1) those of other serum biochemical parameters such as alanine aminotransferase (ALT), aspartate aminotransferase (AST) and gamma glutamyl transpeptidase (GGT), and bilirubin concentrations; and (2) criteria representative of liver histological status. RESULTS: Compared with healthy subjects, DPPIV activity was significantly increased during viral infections and in PBC (P<0.01). In HCV-infected patients, the median activity (interquartile range, IQR), 29.78 IU/l (24.66-35.95), differed significantly (P<0.05) from that of controls: 21.42 (19.76-24.93). No correlation was observed between DPPIV activity and either ALT, AST, bilirubin, or the stage of liver fibrosis and necroinflammatory activity, although GGT was moderately correlated (r=0.58, P<0.05). CONCLUSIONS: Although we confirmed an elevation of serum DPPIV activity in PBC, it seems to be a non-specific phenomenon common to viral infections. The absence of correlation between serum DPPIV and markers of liver disease in HCV-infected patients, suggests that this activity originates not only from the liver, but also from other sources such as peripheral blood cells involved in the control of viral infections. PMID- 12727529 TI - HBV DNA levels and transmission of hepatitis B by health care workers. AB - BACKGROUND: Laboratory-based study funded by the Research and Development Division of the Department of Health to inform the decision making on guidelines for the conduct of exposure prone procedures (EPPs) by health care workers who are hepatitis B carriers. OBJECTIVES: Define the quantity and nature of hepatitis B virus (HBV) DNA in hepatitis carriers whose serum does not contain hepatitis B e antigen (HBeAg) and in surgeons previously cleared to conduct EPPs who have transmitted HBV to their patients. STUDY DESIGN: Cross-sectional survey using HBV DNA quantification, genotyping and sequencing comparing transmitting surgeons and asymptomatic carriers. RESULTS: HBV DNA could be detected and quantified in 64.5% (136 of 211) of carriers whose serum did not contain HBeAg with a median level 3.6 log(10) copies/ml (range of 5.7 log(10) copies). Pre-core mutation appeared not to affect the HBV DNA level, however, all surgeons carried codon 28 variants and transmitted these variants to their patients. The lowest HBV DNA level in a transmitting surgeon was 4 x 10(4) copies/ml. CONCLUSIONS: Pre-core mutations are common in carriers whose serum does not contain HBeAg and do not specifically identify carriers whose HBV DNA levels are high. It was possible to define a level of virus above which transmission of hepatitis B during conduct of EPPs could not be excluded. PMID- 12727531 TI - A novel DNA virus (SEN) among patients on maintenance hemodialysis: prevalence and clinical importance. AB - BACKGROUND: A recently discovered DNA virus (SEN) has been assumed to be responsible for posttransfusion hepatitis in humans. Phylogenetic analysis of SEN virus has revealed the existence of 8 different strains. Two of them (SEN virus strain H (SENV-H) and SENV-D) have been described as possible candidate viruses for inducing posttransfusion hepatitis. Until now, it is unclear whether patients on maintenance hemodialysis are on increased risk for acquiring SEN virus. OBJECTIVES: To investigate the prevalence of SENV-H among patients on maintenance hemodialysis and to examine whether special measures have to be taken to prevent nosocomial spreading of the virus. STUDY DESIGN: Serum samples derived from 78 chronically hemodialysed patients were examined for SENV-H viremia by seminested polymerase chain reaction. A panel of 226 samples from healthy blood donors served as a control group. RESULTS: The prevalence of SENV-H was determined to be 12.8% (n=10) among patients on maintenance hemodialysis. This is nearly the same prevalence as in healthy blood donors (16.8%; n=38). None of the solely SENV-H viremic individuals had clinical or biochemical signs of liver disease. Enhanced severity of liver disease could not be observed in patients coinfected with hepatitis C virus and SENV-H. CONCLUSION: We conclude that SENV-H viremia is widespread among hemodialysis patients. Since no viremic patient had clinical or biochemical signs of liver disease, in our setting the hepatitis-inducing capacity of SENV-H remains unclear. On the basis of our results, at present, we do not regard it as necessary to dialyse SENV-H-viremic patients on separate machines. PMID- 12727532 TI - Are waterborne astrovirus implicated in acute digestive morbidity (E.MI.R.A. study)? AB - BACKGROUND: With rotavirus and Norwalk-like viruses, astroviruses are now recognized as important etiologic agents of viral gastroenteritis in all age groups. However, astrovirus is neither routinely screened for in stool samples, nor in environmental samples, and data on the health impact of waterborne astrovirus are lacking. OBJECTIVES: To assess the potential impact of astrovirus in drinking water on the incidence of acute digestive conditions (ADC) among a panel of volunteers. STUDY DESIGN: The Epidemiology and MIcrobial Risk Assessment (E.MI.R.A.) study combined a daily epidemiological follow-up of digestive morbidity among a panel of 544 volunteers supplied by French public water systems, and a microbiological surveillance of drinking water. Cases of digestive morbidity were collected through weekly telephone calls. The bacterial, virological and parasitic quality of tap water was assessed monthly. Additional samples were collected if the incidence of ADC increased. The relationship between incidence of ADC during a 7-day period centered about the water sampling day and astrovirus RNA prevalence in drinking water was modeled by regression techniques, taking into account several confounders. RESULTS: 12% (8/68) of the analyzed water samples were positive for astrovirus, and presence of astrovirus RNA was associated with a significant increased risk of ADC: RR=1.51 (95% CI=[1.17-1.94], P value=0.002). CONCLUSIONS: This result suggests a role for waterborne astrovirus in the endemic level of digestive morbidity in the general population. Perhaps astrovirus is a candidate test target for viral surveillance of drinking water. PMID- 12727533 TI - Multicentre Italian Study Group (MISG) for the standardisation of hepatitis C virus (HCV) PCR. AB - BACKGROUND: Several studies on standardisation of NAT assays for diagnosis of hepatitis C virus (HCV) infection have been carried out in European countries. In fact the widespreading use of nucleic acid amplification technology (NAT) in diagnostic centres for the evaluation of the HCV infection, requires the application of reference external standards to control laboratory performance; but up to date they are not routinely used. OBJECTIVES: Fifteen diagnostic centres of major Italian Hospitals participated to a quality control study for the standardisation of polymerase chain reaction (PCR)-based HCV-RNA detection, organised by the Committee for the Study of Biotechnology (CoSBio) of the Italian Society of Clinical Microbiology (AMCLI). All the participant centres (PC) used commercial assays, automated or semi-automated. STUDY DESIGN: The study was performed in four rounds. Altogether each centre received 14 reference negative and 22 reference positive sera. The range of copies number per ml of the reference positive sera was 10(4)-10(7). RESULTS AND CONCLUSIONS: Considering the 540 samples tested, 4.54% of false negative (FN) and 4.28% of false positive (FP) results were reported. Thereafter the sensitivity and the specificity were 95.65 and 95.89%, respectively. The errors were distributed among seven out of the 15 PCs. The percentage of FP results was uniformly distributed in each shipment, whereas FN results emerged with the sera at lower HCV genome copies number. The analysis of the data obtained suggests that FP as well as FN results may be attributable to errors or to others problems of laboratories. To improve the performance of Italian, as well as of laboratories throughout the world, the use of external reference standards in multicentre collaborative studies will be required. PMID- 12727534 TI - The use of simple, rapid tests to detect antibodies to human immunodeficiency virus types 1 and 2 in pooled serum specimens. AB - BACKGROUND: The use of pooled specimens has been proposed as a means of expanding testing for human immunodeficiency virus (HIV) antibodies in population studies and in blood screening, while reducing laboratory costs. OBJECTIVES: To develop a strategic specimen pooling method to be used with rapid HIV antibody assays to detect positive specimens and to evaluate its performance in comparison with testing with commercial EIA and WB. STUDY DESIGN: Two lateral flow rapid HIV antibody assays, Seroz*Strip HIV-1/2(1) and Determine HIV-1/2, were evaluated for their ability to detect HIV-1 antibodies in serum and/or plasma specimens pooled in sizes ranging from two to 20 following the respective manufacturers' protocols. One thousand prospectively collected specimens and 55 seroconversion specimens were prepared in pools of five for evaluation by the two rapid HIV assays. RESULTS: Optimal detection and discrimination of HIV-1 antibody-positive and HIV-1 antibody-negative specimens was observed in pool sizes of five to ten for both assays. The ability of the two rapid assays to detect HIV-1 antibody positive samples from commercial HIV-1 seroconversion panels contained in the pools was equivalent to that of commercial enzyme immunoassays (EIAs) and Western blot (WB) to detect HIV-1 antibody in the non-pooled samples. Application of the pooling method in prospectively collected specimens yielded excellent concordance with EIA/WB results in both sensitivity (98.88% for Seroz*Strip HIV-1/2, 100% for Determine HIV-1/2) and specificity (99.56% for Seroz*Strip HIV-1/2, 99.45% for Determine HIV-1/2). CONCLUSION: Use of a pooling strategy with either assay reduced the number of tests required by almost 50% and could provide substantial cost reductions for HIV screening in settings where HIV-1 prevalence is less than 10%. PMID- 12727535 TI - Assessment of precision and concordance of quantitative mitochondrial DNA assays: a collaborative international quality assurance study. AB - BACKGROUND: A number of international research groups have developed DNA quantitation assays in order to investigate the role of mitochondrial DNA depletion in anti-retroviral therapy-induced toxicities. OBJECTIVES: A collaborative study was undertaken to evaluate intra-assay precision and between laboratory concordance of measurements of mitochondrial DNA quantity, as a component of a comprehensive quality assurance project. STUDY DESIGN: Four laboratories were asked to measure and report mitochondrial DNA and nuclear DNA genome copy number, as well as mitochondrial DNA copy number/cell, for 17 coded aliquots of DNA derived from serial dilutions of pooled DNA from a lymphoblastoid cell line. Samples included masked replicates and five standards. All samples had similar mitochondrial DNA/nuclear DNA ratios. Precision within laboratories was assessed by determining the coefficient of variation of replicates. Concordance between laboratories was assessed by determining the average coefficient of variation of the mean replicate values for each sample. The effect of standardising the assay for these three measurements was also assessed for laboratories A, B and C. RESULTS: Measurements of mitochondrial DNA and nuclear DNA content for replicate samples varied by an average of less than 6% (based on log(10) values, 72% non-logged values), and measurements of mitochondrial DNA/cell for replicates varied by less than 12% (based on log(10) values, 32% non logged values), with no improvement of precision after standardisation. Standardisation did significantly improve the concordance of results for measurements of mitochondrial DNA content and mitochondrial DNA/cell. Non standardised measurements of mitochondrial DNA content for the same sample set varied by 19% between laboratories (based on log(10) values, 96% non-logged values), and after standardisation results varied by less than 3% (based on log(10) values, 54% non-logged values). There was no significant improvement for concordance of measures of nuclear DNA content after standardisation, with results varying by 4.56% between laboratories (based on log(10) values, 45% non logged values) before standardisation, and by 2.49% (based on log(10) values, 50% non-logged values) after standardisation. Derived values of mitochondrial DNA/cell varied between laboratories by an average of 91% (non-logged, 56% log(10) values) before and by 56% (non-logged, 13% log(10) values) after standardisation. CONCLUSION: All assays demonstrated good precision. The use of common standards is an important step in improving the comparability of data between laboratories. PMID- 12727536 TI - Lamivudine and Famciclovir resistant hepatitis B virus associated with fatal hepatic failure. AB - BACKGROUND: Lamivudine (LMV) is the only nucleoside analogue approved for the treatment of chronic hepatitis B (CHB). LMV, as with other nucleoside analogues including Famciclovir (FCV), suppresses the replication of hepatitis B virus (HBV) by targeting the viral polymerase. However, prolonged antiviral therapy results in the emergence of drug resistance HBV which can contribute to virological breakthroughs and recurrent hepatitis flares. OBJECTIVES: A 38-year old hepatitis B e antigen (HBeAg) positive Chinese female infected with genotype B HBV commenced treatment with FCV and LMV combination therapy but was later maintained on LMV monotherapy. The patient remained HBeAg positive throughout treatment. Virological breakthrough occurred with the emergence of drug resistant HBV. This coincided with worsening liver function and the patient died of subacute fulminant hepatitis. This study evaluated the virological factors that contributed to the clinical decline of the patient. STUDY DESIGN: Biochemical analysis and full-length HBV genomic sequencing were performed on serial serum samples collected from the patient before and during antiviral therapy. RESULTS: Virological analysis revealed that the pre-treatment dominant HBV quasispecies in the patient had a number of non-consensus genotype B mutations which were located in the basal core promoter (BCP), polymerase, X, core and S genes. Subsequent to the instigation of antiviral therapy, the dominant drug resistant HBV which caused virological breakthrough and was associated with hepatic failure displayed a series of unique mutations particularly in the BCP (A1762T and G1764A) and in the polymerase (rtL180M, rtM204V, rtA222T and rtL336V), core (cP5T, cS26A, cV85I and cP135A), surface (sI195M and sM213I) and X (xK95Q, xN118T, xK130M and xV131I) proteins. CONCLUSIONS: Monitoring for the accumulation of unique mutations within the genome of drug resistant HBV mutants isolated during long term antiviral therapy appears warranted in the clinical management of patients with CHB. PMID- 12727537 TI - Four simple procedures for the assay of methdilazine in bulk drug and in tablets and syrup using potassium iodate. AB - Four simple, selective, accurate and reproducible procedures are described for the assay of methdilazine in bulk form and in formulations. One titrimetric and three spectrophotometric methods are based on the oxidation of the drug with potassium iodate, and determination of either excess iodate or iodine released in the reaction. In the titrimetric method (Method A) the drug is reacted with a known excess of iodate in sulphuric acid medium followed by the iodometric determination of residual oxidant. The residual oxidant is determined by reacting it with variamine blue and measuring the absorbance of the oxidised dye at 540 nm (Method B). The second spectrophotometric method (Method C) is based on the oxidation of the drug in sulphuric acid medium in the presence of chloride ions by a large excess of iodate and the iodate being reduced to iodine. The ICl(2)(-) generated in this reaction is used to iodinate 2',7'-dichlorofluorescein dye, and the red colour of the iodinated dye is measured at 525 nm. The other spectrophotometric method (Method D) also involves the oxidation of the drug in acid medium by a large excess of iodate with the liberation of iodine and its subsequent extraction with carbon tetrachloride followed by measuring the absorbance 520 nm. The methods were successfully applied to the determination of methdilazine in tablets and syrup and the results obtained in agreement with the label claim. PMID- 12727538 TI - Spectrophotometric determination of labetalol in pharmaceutical preparations and spiked human urine. AB - Two simple and sensitive spectrophotometric methods were developed for the spectrophotometric determination of labetaolol (LBT). Both methods are based on the phenolic nature of the drug. The first method (Method I) is based on coupling LBT with diazotized benzocaine in presence of trimethylamine. A yellow colour peaking at 410 nm was produced and its absorbance is linear with the concentration over the range 1-10 microg ml(-1) with correlation coefficient (n=5) of 0.9993. The molar absorptivity was 2.633 x 10(4) l mol(-1) cm(-1). The second method (Method II) involves coupling LBT with diazotized p-nitroaniline in presence of sodium carbonate. An orange colour peaking at 456 nm was obtained and its absorbance is linear with concentration over the range 1-10 microg ml(-1) with correlation coefficient (n=5) of 0.99935. The stoichiometry of the reaction in both cases was accomplished adopting the limiting logarithmic method and was found to be 1:1. The developed method could be successfully applied to commercial tablets. The results obtained were in good agreement with those obtained using the official methods. No interference was encountered from co-formulated drugs, such as hydrochlorothiazide. The method was further extended to the in-vitro determination of LBT in spiked human urine. The % recovery (n=4) were 97.7+/-5.75 and 103.27+/-5.42 using the Methods I and II, respectively. PMID- 12727539 TI - New substituted 2-(pyrazol-1-yl)-dialkylacetanilides with potential local anesthetic and antiarrhythmic action. Part II. AB - Nine substituted 2-(pyrazol-1-yl)-dialkylacetanilides were synthesized by N alkylation of pyrazole and its derivatives with several 2-iodoacetanilides. The new compounds exhibited local anesthetic and antiarrhythmic actions. They have been characterized by elemental chemical analysis, UV-Vis, IR, 1H NMR, 13C NMR, SM spectra and pharmacology research. PMID- 12727540 TI - Kinetics of cefamandole nafate degradation in solid phase. AB - The influence of temperature and relative humidity (RH) on the stability of cefamandole (CM) nafate sodium in the solid phase was investigated. Changes in the concentration of cefemandole nafate sodium were recorded using HPLC with UV detection. The method was validated for the following parameters: selectivity, linearity, precision, limit of detection and sensitivity. It showed good linearity (r=0.9996) in the range 0.4 x 10(-4)-5.6 x 10(-4) g ml(-1) using a LiChrospher RP-18 column and as mobile phase acetonitryle-triethylamine (10% v/v, adjusted to pH 2.5 with phosphoric acid (84%) and diluted with water) (35:65). The degradation of CM occurring at 0% RH of the ambient air and at air humidity RH>50% is a first-order reaction relative to substrate concentration. The first order rate constants (k) were determined for CM degradation in dry air at 373, 383, 388 and 393 K, at air humidity RH=76.4% at 323, 333, 343 and 353 K, and at 353 K at air humidity RH>50%. The kinetic and thermodynamic parameters of the decomposition were calculated. PMID- 12727541 TI - Synthesis and antifungal activity of (+/-)-1-(5-aryl-3-pyridin-2-yl-4,5-dihydro pyrazol-1-yl)-2-imidazol-1-yl-ethanone derivatives. AB - (+/-)-1-(5-Aryl-3-pyridin-2-yl-4,5-dihydro-pyrazol-1-yl)-2-imidazol-1-yl-ethanone derivatives were synthesized and tested for their in vitro antifungal activity. The compounds showed a moderate activity against strains of Candida parapsilosis, Candida pseudotropicalis and Candida glabrata. PMID- 12727542 TI - Synthesis and anti-inflammatory activity of 5-(6-methyl-2-substituted 4 pyrimidinyloxymethyl)-1,3,4-oxadiazole-2-thiones and their 3-morpholinomethyl derivatives. AB - The synthesis of 5-(6-methyl-2-substituted 4-pyrimidinyloxymethyl)-2,3-dihydro 1,3,4-oxadiazole-2-thiones and their 3-morpholinomethyl derivatives and the results of anti-inflammatory activity in vivo are described. Most of the tested compounds exhibited anti-inflammatory activity and some of them were more active than acetylsalicylic acid. PMID- 12727543 TI - Synthesis of new P3CS derivatives and their mitogenic activity on in vitro mice splenocytes. AB - Vaccination against tumors represents a relevant issue in current human cancer therapy. The N-terminal part of the lipoprotein from the outer membrane of Escherichia coli, tripalmitoyl-S-glyceryl-Cys-Ser (P(3)CS) and analogs with longer aminoacidic sequence are polyclonal activators for B-lymphocytes. Previous study reported that their N-2,2,2-trichloroethoxycarbonyl (Troc) derivatives increase immunocyte mitogenic activity. Therefore, in order to obtain compounds of greater activity and to investigate relationships between molecular structure of S-glyceryl skeleton and biological activity, we synthesized new Troc derivatives of P(3)CS. The mitogenicity of compounds was determined in vitro, by measuring in vitro [3H]-thymidine incorporation into splenocytes from Balb/c mice. Concentrations of compounds ranged from 0 to 64 micro g/ml. In particular, S-[2,3-bis(trichloroethoxycarbonyloxy)]-N-trichloroethoxycarbonyl dipeptide derivative exhibited significant mitogenic activity endowed with high pharmacological potency. These new series of compounds could be used as potent immunoadjuvants for the development of novel synthetic vaccines for tumor immunotherapy. PMID- 12727544 TI - Kinetic study of alkaline induced hydrolysis of the skeletal muscle relaxant chlorzoxazone using ratio spectra first derivative spectrophotometry. AB - 2-Amino-4-chlorophenol was found to be the alkaline induced degradation product and the synthetic precursor of chlorzoxazone. The aim of this work is to study different factors affecting the degradation process due to the high toxicity of 2 amino-4-chlorophenol. Chlorzoxazone was found to follow pseudofirst order kinetics. Ratio spectra first derivative spectrophotometry (DR(1)) was developed for monitoring the change in chlorzoxazone concentration during the degradation process. Kinetic parameters (rate constant (K) and half-life (t(0.5))) were calculated at different temperatures (40-120 degrees C) and different sodium hydroxide concentrations (3-10 M). Activation energy at 3 and 8 M sodium hydroxide concentration and alkaline induced catalysis constant at 60, 70 and 80 degrees C were also calculated. PMID- 12727545 TI - Environmental endocrine disrupters dysregulate estrogen metabolism and Ca2+ homeostasis in fish and mammals via receptor-independent mechanisms. AB - Xenoestrogen endocrine disrupters (EDs) in the environment are thought to be responsible for a number of examples of sexual dysfunction that have recently been reported in several species. There is growing concern that these compounds may also cause abnormalities of the male reproductive tract and reduced spermatogenesis in man. Whilst some effects of EDs may be receptor-mediated, there is growing evidence that these compounds can exert potent effects in vivo by directly interacting with cellular enzyme targets. Here we report on, and review, the effects of alkylphenols and other EDs on two such enzymes: (1) sulfotransferases, which convert active estrogenic steroids to inactive steroid sulfates; and (2) Ca(2+)-ATPases, which are responsible for maintaining low, physiological, intracellular Ca(2+) concentrations. These enzymes are potently inhibited by EDs in both fish and mammalian species. The increased concentrations of active estrogens and the likely cytotoxic effects of elevated concentrations of intracellular Ca(2+) arising from these effects may underlie some of the endocrine disrupting potential of these widespread industrial pollutants. PMID- 12727546 TI - Nitrite disrupts multiple physiological functions in aquatic animals. AB - Nitrite is a potential problem in aquatic environments. Freshwater fish actively take up nitrite across the gills, leading to high internal concentrations. Seawater fish are less susceptible but do take up nitrite across intestine and gills. Nitrite has multiple physiological effects. Its uptake is at the expense of chloride, leading to chloride depletion. Nitrite also activates efflux of potassium from skeletal muscle and erythrocytes, disturbing intracellular and extracellular K(+) levels. Nitrite transfer across the erythrocytic membrane leads to oxidation of haemoglobin to methaemoglobin (metHb), compromising blood O(2) transport. Other haem proteins are also oxidised. Hyperventilation is observed, and eventually tissue O(2) shortage becomes reflected in elevated lactate concentrations. Heart rate increases rapidly, before any significant elevations in metHb or extracellular potassium occur. This suggests nitrite induced vasodilation (possibly via nitric oxide generated from nitrite) that is countered by increased cardiac pumping to re-establish blood pressure. Nitrite can form and/or mimic nitric oxide and thereby interfere with processes regulated by this local hormone. Steroid hormone synthesis may be inhibited, while changes in ammonia and urea levels and excretion rates reflect an influence of nitrite on nitrogen metabolism. Detoxification of nitrite occurs via endogenous oxidation to nitrate, and elimination of nitrite takes place both via gills and urine. The susceptibility to nitrite varies between species and in some cases also within species. Rainbow trout fall into two groups with regard to susceptibility and physiological response. These two groups are not related to sex but show significant different nitrite uptake rates. PMID- 12727548 TI - Physiology of the alveolar surface network. AB - The alveolar surface network (ASN) is the totally fluid intraacinar conformation of the alveolar surface liquid (ASL) continuum circulating, both in series and in parallel, through ultrathin (to <7 nm) molecular conduits formed by appositions of unit bubbles of alveolar gas. The ASN is the analogue of foam in vitro. Appositions of unit bubble films, namely foam films, include (a) bubble-to-bubble at the alveolar entrance, across alveolar ducts, and at pores of Kohn ('classical foam films'); (b) bubble-to-epithelial cell surface ('cell-surface foam film'); and (c) bubble-to-open surface liquid layer of the terminal conducting airways ('surface foam film'). These appositions of monolayer bubble films create (a) 'macrochannels' ('pressure points', 'reservoirs') that modulate ASL transfers, volume and flow throughout the acinus and between acinar surface and both the interstitium and the terminal conducting airways surfaces, and (b) 'microchannels' along the broadest surfaces of the appositions. 'Microchannels', which are expectedly bilayer, serve several functions, including (a) virtually frictionless orientation of unit bubbles and ASL to fill the acinar air space; (b) virtually unrestricted diffusion of respiratory gases; (c) architectural support ('infrastructure') against the 'mass' and 'recoil' force of the interstitium; and (d) provision of 'gate' and 'bridge' dynamics that further modulate and direct ASL circulation. The physiological and anatomical boundary between acinar ASN and the bubble-free open liquid surfaces of the conducting airways is marked by the surface foam film. The ASN operates as outlined above in all regions of the lung, at all lung volumes, beginning at the onset of air breathing at birth and continuing throughout life. Reports of its discovery (Pulmonary Physiology of the Fetus, Newborn and Child (1975) 116; Pediatr. Res. 12 (1978) 1070) and subsequent confirmatory research including the adult lung are summarized in this review by progressive development of each function. These functions, which are normal for a relatively dry foam such as the ASN (where gas:liquid volume ratio is >99:1) cannot be duplicated by the conventional theories and models of an open 'alveolar lining layer'. The unfortunate research technologies upon which these theories and models have been formulated have, indeed, obfuscated recognition of the ASN in vivo. They are also presented and critiqued in this review. PMID- 12727547 TI - Chronic effects of copper exposure versus endocrine toxicity: two sides of the same toxicological process? AB - Chronic sub-lethal exposure to copper (Cu) causes a series of cellular and physiological changes in fish that enable the animal to survive. Copper is also an endocrine disrupting metal in the aquatic environment, and has a number of normal neuro-endocrine roles in vertebrates. This paper explores whether the chronic effects of Cu exposure can be explained by the effects of Cu on neuro endocrine functions in fish. Chronic Cu exposure involves complex physiological adjustments in many body systems, including increased oxygen consumption, reduced mean swimming speed, up-regulation of ionic regulation, decreasing lymphocyte levels and increasing neutrophils, altered immunity, modulation of Cu-dependent and independent enzyme activities, and proliferation of epithelial cells in gills or intestine. These responses can occur with exposure via the food or the water and can be rationalised into three major categories: (1) up-regulation of enzymes/metabolism (2) altered haematopoietic responses and (3) altered cellularity (cell type, turnover or size) in tissues. Some of these responses can be explained by stimulation of general stress responses, including the adrenergic response and stimulated cortisol release via the hypothalamic-pituitary interrenal axis. This can occur despite evidence of vacuolation and foci of necrosis in the brain, and increased macrophage activity, in the kidney of fish exposed to dietary Cu. In addition to generic stress responses, Cu regulates specific neuro-endocrine functions, including the loss of circadian rhythm during dietary Cu exposure that involves the failure to respond to circulating melatonin and a loss of circulating serotonin. We conclude that the chronic physiological effects of Cu and apparent endocrine disrupting effects of Cu are two sides of the same toxicological process. PMID- 12727549 TI - Thyroid rhythm phenotypes and hominid evolution: a new paradigm implicates pulsatile hormone secretion in speciation and adaptation changes. AB - Thyroid hormones (THs, T(3)/T(4)) are essential central regulators that link many biological tasks, including embryonic and post-natal growth, reproductive function, and the behavioral and physiological responses to stress. Recently I proposed a novel theory to explain the role of THs in vertebrate evolution. Here I review the concept and discuss its ability to explain changes over time in hominid morphology, behavior and life history. THs are produced in a distinctly pulsatile manner and appear to generate species-specific TH rhythms with distinct ontogenic shifts. Individual variations in genetically controlled TH rhythms (TR phenotypes) must generate coordinated individual variation in morphology, reproduction and behavior within populations. Selection for any manifestation of a particular TR phenotype in an ancestral population selects all traits under thyroid control, resulting in rapid and well-coordinated changes in descendants. The concept provides the first really plausible explanation for a number of phenomena, including the convergent evolution of bipedalism in early hominids, species-specific sexual dimorphism, coordinated changes in morphology, brain function and gut length over time in hominids, cold adaptation in Homo neanderthalensis, the possible independent evolution of H. sapiens in Asia, and regional adaptation of hominid populations. This new paradigm provides a unique theoretical framework for explaining human origins that has important implications for human health. PMID- 12727550 TI - Understanding cardiovascular physiology in zebrafish and Xenopus larvae: the use of microtechniques. AB - Zebrafish and Xenopus, genetically accessible vertebrates with an externally developing, optically clear embryo, are ideally suited for in vivo functional dissection of the embryonic development of the circulatory system. Physiological characterizations of the cardiovascular system are still imperative for a more complete understanding of the connections between genetic/epigenetic factors and cardiovascular development. Here, we review experimental tools and methods that have been developed to measure numerous cardiovascular parameters in these millimetre-sized animals. PMID- 12727551 TI - Effect of temperature on lipids, proteins, and carbohydrates levels during development from egg extrusion to juvenile stage of Cherax quadricarinatus (Decapoda: Parastacidae). AB - The influence of temperature on biochemical composition, survival and duration of development of Cherax quadricarinatus from egg extrusion to juvenile was analyzed. Berried females were individually subjected to each of 22, 25, 28 and 31 degrees C (n=5 per temperature). Egg samples were obtained every 3 days from egg extrusion to juvenile stage for biochemical analysis. Duration of development and survival decreased with increasing temperature. At 22 and 25 degrees C half of the initial lipid content was consumed during development. At 28 and 31 degrees C, 80% of the initial amount of lipids was consumed. For proteins, depletion rate was significantly lower at 25 degrees C (36% of the initial amount) than at 22, 28 and 31 degrees C (61-65% of the initial amount). For carbohydrates, a significant consumption was observed only at 22 degrees C. Total energy consumption was lower at 22 and 25 degrees C than at 28 and 31 degrees C. We conclude that 22-25 degrees C is the optimal temperature range for C. quadricarinatus egg incubation, although 25 degrees C might be better in terms of development duration in terms of survival, energy cost and protein consumption. PMID- 12727552 TI - Effect of blood withdrawal and angiotensin II on prolactin release in the tilapia, Oreochromis mossambicus. AB - Repeated blood withdrawal (5% of estimated blood volume at 0, 1, 4, 8, 24, 48 and 76 h) from tilapia acclimated to fresh water (FW) resulted in a marked increase in plasma levels of prolactin (PRL) during the first 8 h, reaching a peak above 300 ng/ml after 4 h. The increase in plasma PRL levels was significant except for the level after 72 h. A slight but significant decrease in plasma osmolality was observed at all time points after the blood withdrawal. Repeated blood withdrawal from fish acclimated to seawater (SW) resulted in a marked increase in plasma osmolality after 4 and 8 h. A significant increase was observed in plasma growth hormone (GH) in the fish in SW until the end of the experiment, but there was no change in plasma PRL. Plasma levels of cortisol were significantly higher in the fish in SW than in those in FW during the first 24 h. Blood withdrawal resulted in a significant reduction in hematocrit values in both FW- and SW-adapted fish, suggesting hemodilution. In a separate experiment, a single blood withdrawal (20% of total blood) stimulated drinking after 5 h, regardless of whether the fish were held in FW or SW. Plasma PRL level was also elevated following a single blood withdrawal in the fish acclimated to FW, but not in the fish in SW. Intraperitoneal injection of ANG II (1.0 microg/g) into the fish in FW significantly increased plasma PRL levels after 1 h. Activation of the renin angiotensin system after blood withdrawal and the dipsogenic action of angiotensin II (ANG II) are well established in fish. The reduction in plasma osmolality after repeated blood withdrawal in FW and the increased osmolality in SW suggest that blood volume is restored, at least in part, by drinking environmental water. These results suggest that the marked increase in PRL concentration after blood withdrawal from the fish in FW is due, at least in part, to a facilitative effect between ANG II and reduced plasma osmolality. PMID- 12727553 TI - Apparent diffusion limitations on branchial CO2 transfer are revealed by severe experimental anaemia in brown bullhead (Ameiurus nebulosus). AB - In this study, the rapid (within 2 h) effects of acute anaemia on blood gas and acid-base status, as well as cardiorespiratory variables, were examined in brown bullhead (Ameiurus nebulosus). Anaemia was induced by blood withdrawal coupled to volume replacement with saline. Lowering haematocrit from the control value of 23.5+/-1.0% (mean+/-S.E.M.; N=37) to 5.9+/-0.3% (N=37) resulted in a significant increase (by 2.63+/-0.51 torr; N=7) in arterial CO(2) tension (PaCO(2)) over the subsequent 2-h period in the absence of a change in arterial O(2) tension (PaO(2)). Treatment with bovine carbonic anhydrase (CA) reduced the extent of the PaCO(2) increase to the point where it was not statistically significant. In both control and CA-treated fish, arterial pH decreased during acute anaemia; the acidosis was of mixed respiratory and metabolic origin in control fish and primarily metabolic in CA-treated fish. Inducing anaemia caused increases in both cardiac output (V*b) and heart rate that were similar in control and CA-treated fish. Experimental elevation of V*b equivalent to that observed during anaemia, but in the absence of lowered haematocrit, increased PaCO(2) significantly by 1.49+/-0.74 to 1.64+/-0.78 torr (N=5) without affecting PaO(2). These findings suggest that CO(2) excretion in bullhead, as in rainbow trout, is effectively diffusion-limited, and that approximately half of the increase in PaCO(2) measured during the initial 2 h of anaemia results from the impact of increased blood flow (hence decreased gill transit time) in a diffusion-limited system. PMID- 12727554 TI - Inflammatory agonist stimulation and signal pathway of oxidative burst in neonatal chicken heterophils. AB - Heterophils are the predominant polymorphonuclear leukocytes (PMNs) in poultry. The oxidative burst of activated heterophils, which generates reactive oxygen species (ROS), is one of the first line cellular defenses against invading microorganisms. In this report, the oxidative response of heterophils from neonatal chicks to in vitro stimulation by various inflammatory agonists was investigated using a fluorescence microplate assay. Both non-opsonized formalin killed Salmonella enteritidis and Staphylococcus aureus were able to stimulate heterophil oxidative burst. The phorbol myristate acetate (PMA) was the most potent stimulant for the chicken heterophil oxidative response, whereas, the bacterial cell surface components lipopolysaccharide (LPS) and lipoteichoic acid (LTA) were less effective. Protein kinase C (PKC) is an essential signaling component regulating heterophil oxidative response to stimulation by PMA, LPS, LTA and S. enteritidis. However, inhibition of PKC did not affect the oxidative response to stimulation by S. aureus, suggesting differential signaling pathway responsible for the activation of oxidative burst by Gram-negative S. enteritidis and Gram-positive S. aureus. Inhibition of mitogen activated protein (MAP) kinase p38 and extracellular response kinase (ERK) by SB 203580 and PD 098059, respectively, did not inhibit activated oxidative burst. PMID- 12727555 TI - Chick metabolic rate and growth in three species of albatross: a comparative study. AB - The relative importance of genetic vs. environmental factors in determining the pattern of avian post-embryonic development is much debated. Previous cross fostering of albatrosses suggested that although inter-specific variation in growth rate was determined primarily by differences in dietary energy content, species-specific constraints might have evolved that could limit maximal growth, even in chicks fed at similar rates and on similar diets. This study aimed to determine whether intrinsic differences in resting metabolic rate were apparent during the linear phase of growth in chicks of three species (black-browed, grey headed and light-mantled sooty albatrosses). There was a gradual increase in absolute, and a reduction in mass-specific metabolic rate from 5.0 W kg(-1) during the earliest part of linear growth, to 3.5 W kg(-1) by the time chicks reached peak mass. These values are considerably higher than in resting adults of comparable or lower mass, presumably reflecting the large size and high metabolic demand of organs involved in rapid nutrient processing and tissue synthesis by chicks. The lack of any detectable inter-specific variation in the pattern of metabolic rate changes casts some doubt on the existence of fundamental differences in growth rate that cannot be attributed simply to differences in dietary energy or nutrient delivery rate. PMID- 12727556 TI - The Longmire I, II, and III operations. PMID- 12727557 TI - Is there a relationship between case volume and survival in breast cancer? AB - BACKGROUND: Several studies have suggested that case volume predicts survival in breast cancer, that patients treated in hospitals with larger case volumes survive longer. The present study is a review of cases from the Blue Mountain Regional Tumor Registry and tests that hypothesis. METHODS: A review was made of 2,409 breast cancer cases accessioned from nine hospitals between 1980 and 1995, tabulating hospital annual case volume, stage at diagnosis, age, treatment, and 5 year relative survival rate. Correlations and probabilities are presented. RESULTS: Survival correlates with stage at diagnosis (P <0.001), but not with hospital case volume (P = 0.40). CONCLUSIONS: If, as in this study, survival correlates with stage at diagnosis and not with case volume, then improving survival requires identifying cases earlier. To do that requires improving saturation of the population at risk with effective screening and improving access to healthcare. That implies dispersing services instead of concentrating them in high-volume centers. PMID- 12727558 TI - The effect of high dehydroepiandrosterone sulfate levels on tamoxifen blockade and breast cancer progression. AB - BACKGROUND: We investigated the stimulatory potential of dehydroepiandrosterone sulfate (DHEA-S) on tamoxifen-treated cells and assessed its effect on cancer progression in the adjuvant setting. METHODS: Mean serial serum levels of sex hormones from 44 patients receiving tamoxifen were correlated with follow-up status. T-47D (ER+/PR+) and HCC1937 (ER-/PR-) breast cancer cells were pretreated with 100 microM anastrozole, with or without tamoxifen, and stimulated with 22.8 microM DHEA-S. Rapid colorimetric assays allowed calculation of growth percent change. RESULTS: Clinically, development of metastatic disease correlated only with > or =90 microg/dL DHEA-S (P = 0.005). In vitro, T-47D cells stimulated with DHEA-S after anastrozole showed 35% increased growth. Addition of 0.01 nM tamoxifen demonstrated -7% inhibition. Increasing the DHEA-S/tamoxifen ratio reversed suppression to +25%. CONCLUSIONS: DHEA-S > or =90 microg/dL is a risk factor for recurrence in the adjuvant setting. In vitro, although tamoxifen inhibits cell growth at high doses it can be circumvented by DHEA-S. These results indicate that DHEA-S contributes to tamoxifen resistance and disease progression. PMID- 12727559 TI - Normal mammography and ultrasonography in the setting of palpable breast cancer. AB - BACKGROUND: Each year thousands of women present to general surgeons with palpable breast masses, some of which are clinically ambiguous and the majority of which are benign. In addition, surgeons are frequently faced with the question of whether to biopsy those palpable abnormalities in the setting of normal radiographic studies. One might propose that such lesions could be safely observed rather than immediately biopsied. If these lesions were not biopsied, how many cancers would escape detection? To address this issue, a population of patients with known, palpable breast cancer was retrospectively examined to determine the frequency of normal or benign findings on both mammography and ultrasonography. METHODS: Between January 1998 and December 2001, 351 women with breast carcinoma presented initially with palpable tumors. The medical records of these remaining 351 cases were retrospectively reviewed to examine the radiographic characteristics of the palpable carcinomas. RESULTS: Of the 351 cases in the study group, 13 (3.7%) patients with palpable breast cancers had mammogram and sonogram examinations that were both normal, benign, or nonspecific in appearance. CONCLUSIONS: The results of this study indicate that nearly 4% of women with breast cancer who present with palpable lumps will have normal or benign findings on both mammography and ultrasonography. These data support prior studies of similar false negative rates and may provide some reassurance to surgeons and patients regarding clinical breast lumps, as the decision of whether to biopsy still rests in the surgeon's hands. However, inappropriate reliance on these tests for an evaluation of a palpable abnormality will result in a number of missed tumors. PMID- 12727560 TI - Predictive value of intraoperative touch preparation analysis of sentinel lymph nodes for axillary metastasis in breast cancer. AB - BACKGROUND: Accurate intraoperative diagnosis of axillary malignancy facilitates completion axillary lymph node dissection (ALND) at the time of initial surgery. The capability to address both the primary tumor and axillary disease in a single procedure offers several advantages. This study was designed to define the predictive value of intraoperative touch preparation analysis of sentinel lymph nodes for axillary metastasis in breast cancer and to evaluate the ability of the technique to facilitate accurate synchronous ALND. METHODS: A consecutive cohort of patients with breast cancer at an Army medical center underwent intraoperative touch preparation analysis of sentinel lymph nodes concordant with initial excision. Those found to have sentinel nodes positive by touch preparation analysis underwent ALND at the initial procedure. Patients with negative sentinel nodes by touch preparation analysis, but positive by final pathology, underwent subsequent ALND. Results of the touch preparation analysis were compared with the final pathology. RESULTS: Over a 16-month period, 71 consecutive patients with breast cancer underwent initial excision and touch preparation analysis of 162 sentinel lymph nodes. Final pathology confirmed axillary metastasis in 32% (23 of 71) of patients. Of these, intraoperative touch preparation analysis identified 48% (11 of 23). There were no false positives or unnecessary axillary dissections based upon touch preparation results. Per sentinel node, the positive predictive value was 100%, the sensitivity was 47%, and the specificity was 100%. On a per patient basis, the positive predictive value was 100%, and the sensitivity and specificity were 48% and 100%, respectively. CONCLUSIONS: Intraoperative touch preparation analysis is an effective adjunct to sentinel lymph node biopsy. In our series, it facilitated a definitive cancer operation at the time of initial surgery in nearly 50% of patients, and ensured that no patient underwent an unnecessary axillary dissection. PMID- 12727561 TI - Accuracy of intraoperative touch imprint cytologic analysis of sentinel lymph nodes in breast cancer. AB - BACKGROUND: Eliminating full axillary dissection (AD) in the face of negative sentinel lymph nodes (SLN) has become increasingly popular. We sought to evaluate the accuracy of intraoperative touch imprint cytology in detecting clinically significant metastatic involvement of the SLN. METHODS: Retrospective review of intraoperative cytology and final hematoxylin-eosin stain evaluation of sentinel nodes examined in a community hospital from 1997 to 2002. RESULTS: During that period 1,585 patients underwent axillary dissection; 380 had SLN identification with 142 patients undergoing intraoperative cytologic evaluation of 446 SLNs. Fourteen patients with negative intraoperative cytology had involvement of a SLN on final hematoxylin-eosin examination, for a sensitivity of 75%, specificity of 100%, negative predictive value of 95%, and a false negative rate of 4.9%. In all but 1 case the metastasis was microscopic and measured 2 mm or less. Seven patients went on to completion axillary dissection (50%) with no additional nodal involvement found. CONCLUSIONS: Intraoperative touch imprint cytology is accurate in predicting clinically significant SLN pathology. Although the false negative rate in this series was 4.9%, in all but 1 case it represented micrometastatic disease that may not necessarily require completion axillary dissection. PMID- 12727562 TI - Treatment of invasive breast carcinoma with ultrasound-guided radiofrequency ablation. AB - BACKGROUND: Radiofrequency ablation (RFA) is a minimally invasive thermal ablation technique. This study reports the safety and efficacy of RFA as a minimally invasive strategy for breast cancers <3 cm diameter in postmenopausal women. METHODS: Twenty-two postmenopausal women (aged 60 years or older) with clinical T-1N0 core biopsy proven breast cancers were studied. Thermocoagulation was undertaken using a sonographically guided RF probe under local anesthesia and sedation. The ablated tumor was resected between 1 and 2 weeks later. Endpoints were technical success, completeness of tumor kill, marginal clearance, skin damage, and patient reports of pain and procedural acceptability. RESULTS: The procedure was well tolerated and cosmesis was excellent. Pathology revealed a central ablation zone surrounded by hyperemia. Coagulative necrosis was complete in 19 of 22 patients. Disease at the ablation zone margin was found in 3 patients and 5 patients had disease distant to the ablation zone consisting of multifocal tumors (2), in-transit metastasis (1), and extensive ductal carcinoma in situ with microinvasive carcinoma (2). Ninety-five percent of patients would be willing to have RFA again. CONCLUSIONS: Radiofrequency ablation can be safely applied in an outpatient setting with acceptable patient tolerance. By itself, RFA cannot be considered effective local therapy. Trials to evaluate RFA complemented with breast irradiation are justified. PMID- 12727563 TI - Is parastomal hernia repair with polypropylene mesh safe? AB - BACKGROUND: Concern over the safety of polypropylene mesh in parastomal hernia repairs has led some to avoid its use. We reviewed our rate of complications and outcomes with polypropylene mesh. METHODS: From January 1988 through May 2002, 58 patients underwent parastomal hernia repair with polypropylene mesh. After closure of the fascia, the stoma was pulled through the center of the mesh, which was placed either above or below the fascia. Multivariate analysis was performed to determine independent predictors for the development of complications. RESULTS: There were 31 end colostomies, 24 end ileostomies, and 3 loop transverse colostomies. Mean follow-up with 50.6 months. Overall complications related to the polypropylene mesh was 36% (recurrence 26%, surgical bowel obstruction 9%, prolapse 3%, wound infection 3%, fistula 3%, and mesh erosion 2%). None of the patients had extirpation of their mesh. Complications were significantly associated with younger age (59.6 versus 67 years, P = 0.04). Cancer patients with stomas had fewer complications (P = 0.02, odds ratio 0.34). Inflammatory bowel disease, stomal type, mesh location, urgent procedures, steroid use, and surgical approaches were not significantly associated with an increased complication rate. Of the 15 patients with recurrence, 7 underwent successful repair for an overall success rate of 86%. CONCLUSIONS: Parastomal hernia repair with polypropylene mesh is safe and effective. PMID- 12727564 TI - A comparison of 50 initial endoluminal endograft repairs for abdominal aortic aneurysm with 50 concurrent open repairs. AB - PURPOSE: To review the results of our initial experience with endovascular repair of abdominal aortic aneurysm (AAA) with respect to morbidity and mortality and to compare these outcomes with those of transabdominal repair. METHODS: We reviewed the first 50 consecutive endovascular AAA repairs performed at our institution from November 1999 to January 2002. Pre-operative risk factors, intraoperative variables and post-operative outcomes were assessed. All endovascular patients were followed with periodic examination, contrast-enhanced computed tomography and/or duplex scanning. Comparison was made to 50 patients undergoing standard open repair over a similar time period. RESULTS: Fifty patients underwent endovascular AAA repair (mean age 72.5, AAA size 5.5 cm). Endovascular devices employed were manufactured by Ancure (Guidant Corp.), and AneuRx (Medtronic). Preoperative risk factors were similar to patients undergoing transabdominal repair. Mean operative time was 169 minutes and estimated blood loss was 450cc with average blood replacement of.18 units. Median ICU stay was 0 days and mean hospital stay was 2.3 nights. There were no conversions to open repair, however there was one aborted endovascular attempt. Morbidity included MI (2%), colon ischemia (1%), acute renal insufficiency (4%) and leg ischemia (4%). There was one death within 30 days. Seven endoleaks were identified (6 type II and 1 type I) and were managed angiographically. CONCLUSIONS: The short-term surgical morbidity and mortality rates for endovascular repair of AAA are acceptably low and are comparable to the transabdominal approach. PMID- 12727565 TI - Intraoperative duplex ultrasonography as an adjunct to technical excellence in 100 consecutive carotid endarterectomies. AB - PURPOSE: To evaluate the relationship of intraoperative duplex ultrasonography (duplex) results to neurologic outcomes and restenosis among patients undergoing carotid endarterectomy (CEA). METHODS: One hundred consecutive CEAs were performed at a military medical center over 28 months. Prospectively acquired demographics, duplex results, revisions, and surgical outcomes were reviewed. RESULTS: Thirty-four percent (34 of 100) of sites were abnormal by completion duplex. Of these, 70% (24 of 34) were B-mode flap type defects located in the common carotid artery (CCA), internal carotid artery (ICA), or external carotid artery (ECA). Twenty-one percent of the defects (7 of 34) were technically unacceptable and immediately revised. Subsequently, 3 perioperative neurologic events occurred, 2 strokes and 1 transient ischemic attack (TIA), all among patients with an identified but unrepaired defect involving the ICA or CCA. This association of unrepaired defect with early stroke or TIA was significant (P = 0.02). No significant association (P >0.05) between unrepaired defects and late ipsilateral stroke or TIA or restenosis was identified. CONCLUSIONS: Intraoperative duplex scanning is a useful adjunct to CEA that can identify correctable mechanical problems. Residual elevated velocities or B-mode flaps 2 mm or greater in the ICA warrant consideration for immediate repair. Findings not requiring revision include flaps <2 mm, as well as isolated ECA defects. Prospectively validated duplex criteria are needed to further define which defects require immediate repair. PMID- 12727566 TI - Effect of emergent presentation on outcome from rectal cancer management. AB - BACKGROUND: We have previously reported outcomes for all rectal cancers in BC in 1996. We found that our local recurrence rates and survival were suboptimal relative to current standards in recent literature. METHODS: In this retrospective, population-based study, we report the influence of emergent presentation (obstruction, perforation, massive hemorrhage) on outcomes, types of surgical procedures and use of staging investigations, and use of adjuvant radiation and chemotherapy. RESULTS: There were 452 invasive adenocarcinomas of the rectum of which 45 were emergent and 407 nonemergent. Disease-specific survival at 4 years for emergent and nonemergent stage II cancers were 66% versus 80%, respectively, and for stage III cancers, 60% versus 73%, respectively (P <0.04). Local recurrence rates at 4 years for emergent and nonemergent stage II cancers were 20% versus 15%, respectively, and for stage III cancers, 70% and 20%, respectively (P <0.05). Surgical resection more frequently involved a stoma for emergent (60%) than for nonemergent (35%) cases (P <0.01). Percent of patients having complete staging investigations were similar between emergent (42%) and nonemergent patients (39%). Adjuvant radiation was given in similar proportion to emergent (61%) and nonemergent (55%) patients. Adjuvant chemotherapy was given to a slightly higher proportion of emergent patients (63%) than nonemergent patients (43%). CONCLUSIONS: We conclude that outcome from rectal cancer management is worse for emergent than nonemergent presentation. Since there is no difference in use of staging investigations or adjuvant therapy, the difference in outcome is likely due to difference in surgical technique between emergent and nonemergent cases. PMID- 12727567 TI - Improvement of laryngopharyngeal reflux symptoms after laparoscopic Hill repair. AB - BACKGROUND: People with proven gastroesophageal reflux disease may also experience symptoms such as voice loss, chronic cough, globus, and sore throat. These laryngopharyngeal reflux symptoms have been reported to respond to prolonged proton pump inhibitor therapy, but the Hill approach to resolving these specific individual symptoms has not been widely reported in surgical literature. METHODS: This clinical outcome study is an analysis of symptom improvement in 145 patients who underwent laparoscopic Hill hiatal hernia repair. A standardized questionnaire was used to score eight gastroesophageal reflux disease symptoms and four laryngopharyngeal reflux symptoms. Also, each patient's primary chief complaints were analyzed. RESULTS: Gastroesophageal reflux and laryngopharyngeal reflux symptoms significantly improved (P < 0.01) compared with preoperative symptoms. Each patient's primary chief complaints improved as well. CONCLUSIONS: This clinical outcome analysis documents symptomatic improvement of laryngopharyngeal reflux and gastroesophageal reflux. Likewise, when these laryngopharyngeal reflux symptoms are chief complaints, with proven gastroesophageal reflux disease, the Hill approach to symptom resolution is likely to be successful. PMID- 12727568 TI - Functional results and quality of life after tracheal resection for locally invasive thyroid cancer. AB - BACKGROUND: Local invasion of the upper aerodigestive tract by thyroid cancer, although uncommon, is a serious cause of morbidity and mortality. The impact of aerodigestive tract resection on the functional status and quality of life of the patient has not previously been investigated. METHODS: Patients with locally invasive thyroid cancer were included in a prospective surgical protocol. Swallowing function was assessed with barium swallow at 7 days and 1 month postoperatively. Postoperative quality of life (QOL) was measured using a validated head and neck QOL instrument. RESULTS: Seven patients underwent airway resection for locally invasive recurrent thyroid cancer in the period 1999 to 2001. At 1 week postoperative 3 of 7 (43%) had no evidence of aspiration on barium swallow. At 4 weeks 6 of 7 (86%) had no aspiration. Postoperative QOL scores in the domains of eating function (85.2) and emotional status (78.6) were significantly better than those of a comparison group undergoing treatment for cancers of the oropharynx, P = 0.012 and P = 0.0077, respectively. CONCLUSIONS: Tracheal resection for locally invasive thyroid cancer is associated with a return to full dietary intake within 4 weeks of surgery in most cases. Function and QOL after this type of surgery are acceptable. PMID- 12727569 TI - Surgeons' anonymous response after bile duct injury during cholecystectomy. AB - BACKGROUND: Bile duct injuries remain one of the most devastating injuries during laparoscopic cholecystectomy. Few studies target surgeons who have experienced bile duct injuries for their insight, their perspective, and their suggestions concerning this problem. METHODS: A confidential questionnaire was sent to all practicing general surgeons under the age of 65 years in British Columbia, Canada. RESULTS: Seventy-five percent of surgeons responded to the survey. Of the 114 questionnaires completed, more than 97% of respondents had completed formal training in laparoscopic cholecystectomy. One half of surgeons reported experience with laparoscopic bile duct injury. A significant difference in years in practice between surgeons with injury and surgeons without injury was noted. The majority of injuries occurred after the surgeons's first 100 cholecystectomies performed. The first thoughts of surgeons after injury uniformly concerned the patient's well being. The next most common thoughts were in relation to obtaining help or a second opinion from another surgeon. Surgeons cited inflammation and short or anomalous cystic ducts as the most responsible factors contributing to injury. The majority of surgeons felt that these injuries are unavoidable and less than half felt that it was always a surgical error. Fewer than 15% thought injuries could be avoided by performing a cholangiogram. Surgeons suggested meticulous dissection and less haste to divide structures may prevent an injury. Surgeons recommend educating colleagues to remove the stigma of failure associated with conversion to laparotomy. CONCLUSIONS: General surgeons in British Columbia have a one in two chance of experiencing a bile duct injury in their career. There were more injuries in surgeons who had already been in practice for 10 years at the time of introduction of laparoscopic cholecystectomy. The injuries are likely to occur despite high volumes of procedures and increased experience. The incidence of bile duct injuries does not seem to be different in surgeons who perform routine cholangiography and most surgeons feel that cholangiography would have little effect on injury incidence. Surgeons tend to have patient-centered concerns after injury and little concern for medicolegal issues. The majority of surgeons felt that these injuries could not be anticipated and as such it is an inherent risk of this procedure. PMID- 12727570 TI - Interferon-based adjuvant chemoradiation therapy after pancreaticoduodenectomy for pancreatic adenocarcinoma. AB - BACKGROUND: Patients with cancer who undergo pancreaticoduodenectomy (PD) followed by radiation and 5-fluorouracil (5-FU) therapy have experienced median overall survival from 18 to 24 months and an actuarial 2-year overall survival from 34% to 48%. We previously reported an 84% 2-year survival using a novel adjuvant chemoradiation protocol that included alpha interferon. This report describes the continued observations regarding this methodology with longer follow-up and more than twice the number of patients as the original report. METHODS: From July 1995 to May 2002, 43 patients with adenocarcinomas in the pancreatic head underwent PD at our institution. The mean age was 62 years (range 29 to 77) and 60% were men. Final pathologic findings were stage I (2%), II (12%), III (72%), and IVa (14%) while 84% had positive lymph nodes (average number of nodes positive was 3.2 nodes, (range 0 to 13). Tumor extended through the capsule of the surgical specimen in 70%. These patients then received our investigational protocol consisting of external-beam irradiation at a dose of 4,500 to 5,400 cGy (25 fractions over 5 weeks) and three-drug chemotherapy: continuous infusion 5-FU (200 mg/m(2) daily, days 1 to 35), weekly intravenous bolus cisplatin (30 mg/m(2) daily, days 1,8,15,22,29), and subcutaneous alpha, interferon (3 x 10(6) units, days 1 to 35). This chemoradiation was followed by continuous infusion 5-FU (200 mg/m(2) daily, weeks 9 to 14 and 17 to 22). Chemoradiation was generally initiated between 6 and 8 weeks after surgery. RESULTS: All patients completed radiation therapy. There were no deaths due to chemoradiation but 42% were hospitalized during chemoradiation, virtually all due to gastrointestinal toxicity. With a mean follow-up time of 31.9 months, 67% of the patients are alive. Therefore, the median survivorship has not been reached. Actuarial overall survival for the 1-, 2-, and 5-year periods was 95% (confidence interval [CI] = 91% to 98%), 64% (CI = 56% to 72%), and 55% (CI = 46% to 65%), respectively. CONCLUSIONS: This follow-up report further suggests overall survival may be improved for patients with adenocarcinoma in the pancreatic head using an adjuvant interferon-based chemoradiation protocol. These results are obtained despite a high incidence of node involvement and advanced tumor stage. From this limited patient series, the actuarial 2-year and 5-year overall survival rates suggest a potential for improved long-term survival. Further study of this regimen in a multiinstitutional setting is needed. PMID- 12727571 TI - Surgical complications of bone marrow transplantation. AB - BACKGROUND: We wished to determine the type of diseases in patients who received bone marrow transplant (BMT) that potentially involve the general surgeon at our institution. METHODS: The records of 542 patients who underwent bone marrow transplant at Oregon Health and Sciences University between January 1990 and December 2000 were retrospectively reviewed. Gastrointestinal complications included in the study were gastrointestinal bleeding, venoocclusive disease of the liver, intestinal graft versus host disease, pneumatosis intestinalis, necrotizing enteritis, as well as other more common surgical diseases (eg, appendicitis). RESULTS: Gastrointestinal complications or surgical consultations were noted in 92 of 542 patients (17%). Of these, formal general surgical consultation was obtained in 48 patients (9%). The most common causes for surgical consult were cholecystitis (5), abdominal pain of unknown etiology (5), central line complications (5), small bowel obstruction (4), and appendicitis (4). Twenty-eight (58%) of these patients received an operation. Six patients (13%) died during the same hospitalization as their surgery consult. Forty-four patients with these gastrointestinal symptoms related to transplantation did not receive surgical consult. The mortality in this group was 45%. CONCLUSIONS: The majority of gastrointestinal complications after bone marrow transplant do not require surgical intervention. However, these conditions may overlap the more common reasons for surgical consult and must be identifiable by the general surgeon. Of patients who did require surgical intervention, it was primarily for common surgical diseases. PMID- 12727572 TI - Serum lactate and base deficit as predictors of mortality and morbidity. AB - OBJECTIVES: To determine whether lactate levels and base deficits in critically ill surgical intensive care unit (SICU) patients correlate and whether either measure is a significant indicator of mortality and morbidity. METHODS: A review was made of 137 SICU patients who had serial lactate and blood gas measurements. Patients were stratified by absolute lactate and base deficit values as well as time to lactate clearance. RESULTS: Initial and 24-hour lactate level was significantly elevated in nonsurvivors versus survivors (P = 0.002). Initial base deficit was not significantly different; 24-hour base deficit did achieve statistical significance (P = 0.02). Subgroup analysis among trauma patients (n = 36) and major abdominal surgery (n = 101) confirmed the significant correlation between lactate levels and survival. There was poor correlation between initial and 24-hour lactate and base deficit among all patients (r = -0.3 and -0.5). Mortality if lactate normalized within 24 hours was 10%, compared with 24% for >48 hours and 67% if lactate failed to normalize. Physical status at discharge was related to initial lactate (P = 0.05), as well as to lactate clearance time (P = 0.01). CONCLUSIONS: Elevated initial and 24-hour lactate levels are significantly correlated with mortality and appear to be superior to corresponding base deficit levels. Lactate clearance time may be used to predict mortality and is associated with outcome at discharge. Initial base deficit is a poor predictor of mortality and did not correlate with lactate levels except in trauma nonsurvivors. In addition to being used as an endpoint for resuscitation, lactate may be predictive of certain morbidities and patient outcome at discharge. PMID- 12727573 TI - Delayed celiotomy for the treatment of bile leak, compartment syndrome, and other hazards of nonoperative management of blunt liver injury. AB - BACKGROUND: Management of blunt liver injury is predominantly nonoperative. However, complications occur in 10% to 25% of patients, with half taking place more than 24 hours after injury. Few reports have addressed the management of the new pattern of these delayed complications, which is the objective of this study. METHODS: Adult patients admitted to our level one trauma center from 1995 to 2000 with blunt liver injury were identified. Demographic, physiologic and laboratory data, computed tomography (CT) and operative findings, and complications were reviewed. RESULTS: Blunt liver injury was identified in 192 patients. Thirty-nine patients (20%) underwent immediate celiotomy. The remaining 153 patients were initially managed nonoperatively. Liver-related complications developed in 19 (12%) patients. Fifteen patients underwent delayed celiotomy to treat secondary inflammatory processes, from bile leak (6), hemorrhage (5), and hepatic abscess (1), and to treat abdominal compartment syndrome (2), and decompress hepatic compartment syndrome (1). Although no deaths or complications were directly caused by delayed celiotomy, 2 deaths (11%), occurring early in this series, were attributed to liver-related complications. CONCLUSIONS: These complications, occurring in 12% of patients with liver injuries, may be a consequence of initial nonoperative management. Although these findings do not negate nonoperative management of blunt liver injury, this approach can be hazardous and requires diligence to recognize and treat delayed and potentially fatal complications. PMID- 12727574 TI - Reinforced silicone elastomer sheeting, an improved method of temporary abdominal closure in damage control laparotomy. AB - BACKGROUND: The ability to massively transfuse and resuscitate critically ill surgical patients has resulted in unprecedented survival and a new set of complications including abdominal compartment syndrome (ACS) and the "unclosable" abdomen. Traditional methods of temporary abdominal closure have met with several limitations, not the least of which is a marked delay in achieving definitive fascial closure. Since 1991, we have consistently used reinforced silicone elastomer (Silastic) sheeting as a form of temporary abdominal closure in these settings. We report our results using this technique in a large cohort of critically ill surgical patients. METHODS: All patients undergoing silicone elastomer temporary abdominal closure since 1991 were identified and their charts abstracted for principal diagnosis and indication for temporary abdominal closure, fluid requirements, number of operations, and time to fascial closure. Time to definitive closure in the respective groups was analyzed using Kaplan Meir survival curves and the Wilcoxon rank-sum test. Odds ratios for death were analyzed using logistic regression. RESULTS: One hundred thirty-four patients underwent temporary abdominal closure with silicone elastomer over this period and only 62% (83) survived their hospital admission. Trauma and ruptured abdominal aortic aneurysm were the most frequent diagnoses. The most frequent indication was edema precluding abdominal closure. The mean crystalloid and blood requirements in the 24 hours preceding temporary abdominal closure were 21 +/- 16 L and 15 +/- 11 U, respectively. Of survivors, 75% (63 of 83) achieved fascial closure during their index admission. The median time to fascial closure in patients ultimately closed was 5 days. The median time to closure and the proportion of patients ultimately closed varied with the indication for closure with an earlier and greater chance of success in patients who could not tolerate closure (ACS) or could not be closed primarily (edema). Age-adjusted mortality was 5 times (95% confidence interval: 2 to 13) higher in patients developing ACS. CONCLUSIONS: Nylon reinforced silicone elastomer is a safe, reliable material for temporary abdominal closure in severely ill patients. Primary fascial closure can be obtained in a timely fashion in the majority of patients. The success of obtaining definitive fascial closure depends on the indication for temporary abdominal closure, with visceral edema and ACS having the highest likeliest of early success. PMID- 12727575 TI - Third heart sound and elevated jugular venous pressure as markers of the subsequent development of heart failure in patients with asymptomatic left ventricular dysfunction. AB - PURPOSE: To determine the independent prognostic value of a third heart sound (S(3)) and elevated jugular venous pressure in patients with asymptomatic left ventricular dysfunction. METHODS: We performed a post hoc analysis of 4102 participants from the Studies of Left Ventricular Dysfunction (SOLVD) prevention trial. In that trial, participants with asymptomatic or minimally symptomatic left ventricular dysfunction (New York Association class I or II, left ventricular ejection fraction < or =0.35, no treatment for heart failure) were allocated randomly to enalapril or placebo and followed for a mean (+/- SD) of 34 +/- 14 months. The presence of an S(3) and elevated jugular venous pressure was ascertained by physical examination at study enrollment. We used multivariate proportional hazards models to determine whether these physical examination findings were associated with the development of heart failure, a prespecified endpoint of the SOLVD prevention trial. RESULTS: At baseline, 209 subjects (5.1%) had an S(3) and 70 (1.7%) had elevated jugular venous pressure. Heart failure developed in 1044 subjects (25.5%). After adjusting for other markers of disease severity, an S(3) was associated with an increased risk of heart failure (relative risk [RR] = 1.38; 95% confidence interval [CI]: 1.09 to 1.73; P = 0.007) and the composite endpoint of death or development of heart failure (RR = 1.34; 95% CI: 1.09 to 1.64; P = 0.005). Elevated jugular venous pressure was also associated with these outcomes in multivariate models. CONCLUSION: The physical examination provides prognostic information among patients with asymptomatic or minimally symptomatic left ventricular dysfunction. PMID- 12727576 TI - Insulin, proinsulin, proinsulin:insulin ratio, and the risk of developing type 2 diabetes mellitus in women. AB - PURPOSE: To assess the associations among baseline levels of fasting insulin and proinsulin, proinsulin:insulin ratio, and the development of type 2 diabetes mellitus in apparently healthy middle-aged women. METHODS: In a nested case control study involving a nationwide cohort of 27,628 participants from the Women's Health Study, 126 women with diabetes diagnosed during a 4-year follow-up period were compared with 225 age-matched controls. Fasting insulin level and proinsulin:insulin ratio were assessed in quartiles, and proinsulin level was assessed in categories (< or =4.0 pmol/L, 4.01 to 6.99 pmol/L, > or =7.0 pmol/L). The risk of developing type 2 diabetes was determined using conditional logistic regression analysis that adjusted for body mass index and other diabetes risk factors. RESULTS: Baseline insulin and proinsulin levels and proinsulin:insulin ratios were significantly higher among cases than among controls. Women with elevated insulin levels in the highest as compared with the lowest quartile were more likely to develop diabetes (odds ratio [OR] = 5.6; 95% confidence interval [CI]: 1.8 to 17.6), as were women with elevated (> or =7.0 pmol/L vs. < or =4.0 pmol/L) proinsulin levels (OR = 16.4; 95% CI: 5.8 to 46.8) and women with proinsulin:insulin ratios in the highest quartile (OR = 9.6; 95% CI: 3.1 to 30.8). Similar results were observed among women with a baseline hemoglobin A(1c) level < or =6.0%. In time-trend analyses, fasting insulin was a consistent predictor of long-term risk. Proinsulin and proinsulin:insulin ratio, although predictive throughout the study, were especially strong predictors of rapid progression to type 2 diabetes. CONCLUSION: Elevated fasting insulin and proinsulin levels and proinsulin:insulin ratio are associated with an increased risk of developing type 2 diabetes in apparently healthy middle-aged women. PMID- 12727577 TI - A randomized trial of aggressive lipid reduction for improvement of myocardial ischemia, symptom status, and vascular function in patients with coronary artery disease not amenable to intervention. AB - PURPOSE: To determine the effects of aggressive lipid lowering on markers of ischemia, resistance vessel function, atherosclerotic burden, and symptom status in patients with symptomatic coronary artery disease. METHODS: Sixty consecutive patients with coronary artery disease that was unsuitable for revascularization were assigned randomly to either usual therapy of lipids for patients with a low density lipoprotein (LDL) cholesterol target level <116 mg/dL, or to a more aggressive lipid-lowering strategy involving up to 80 mg/d of atorvastatin, with a target LDL cholesterol level <77 mg/dL. The extent and severity of inducible ischemia (by dobutamine echocardiography), vascular function (brachial artery reactivity), atheroma burden (carotid intima-media thickness), and symptom status were evaluated blindly at baseline and after 12 weeks of treatment. RESULTS: After 12 weeks of treatment, patients in the aggressive therapy group had a significantly greater decrease in mean (+/- SD) LDL cholesterol level than those in the usual care group (29 +/- 38 mg/dL vs. 7 +/- 24 mg/dL, P = 0.03). Patients in the aggressive therapy group had a reduction in the number of ischemic wall segments (mean between-group difference of 1.3; 95% confidence interval: 0.1 to 2.0; P = 0.04), flow-mediated dilatation (mean between-group difference of 5.9%; 95% confidence interval: 2.5% to 9.4%; P = 0.001), and angina score after 12 weeks. There were no significant changes in atherosclerotic burden in either group. CONCLUSION: Patients with symptomatic coronary artery disease who are treated with aggressive lipid lowering have improvement of symptom status and ischemia that appears to reflect improved vascular function but not atheroma burden. PMID- 12727578 TI - Evidence of autoimmunity in chronic periaortitis: a prospective study. AB - BACKGROUND: Chronic periaortitis includes idiopathic retroperitoneal fibrosis, inflammatory aneurysms of the abdominal aorta, and perianeurysmal retroperitoneal fibrosis. It is considered to be due to advanced atherosclerosis, but is often associated with systemic autoimmune disorders. METHODS: We studied 16 consecutive patients who were diagnosed with chronic periaortitis by computed tomography. Each patient underwent a physical examination, routine laboratory tests, measurement of autoantibodies, thyroid echotomography, and chest radiography. Aortic wall or periaortic retroperitoneal samples from 9 patients who underwent surgery were available for histologic examination and immunohistochemical characterization of the inflammatory infiltrate. RESULTS: Twelve patients had constitutional symptoms, 14 had an elevated erythrocyte sedimentation rate, and 13 had an elevated C-reactive protein level. Antinuclear antibodies were positive in 10 patients. Three patients had autoimmune thyroiditis, and 1 had seropositive rheumatoid arthritis. Antineutrophil cytoplasmic antibodies were positive in 3 patients who presented with rapidly progressive renal failure. Pathologic examination of the aortic and periaortic specimens revealed moderate to severe inflammatory infiltration, mainly consisting of B cells and CD4(+) T cells. Vasculitis with fibrinoid necrosis involving the aortic vasa vasorum and the small and medium retroperitoneal vessels was found in seven of the nine histologic samples. CONCLUSION: These clinical and pathologic features support the hypothesis that, at least in some patients, chronic periaortitis is a systemic autoimmune disease, perhaps involving a vasculitic process of small and medium vessels. PMID- 12727579 TI - Use of a cyclophosphamide-induction methotrexate-maintenance regimen for the treatment of Wegener's granulomatosis: extended follow-up and rate of relapse. AB - PURPOSE: To determine the relapse rate and outcome in patients with Wegener's granulomatosis treated with daily cyclophosphamide and glucocorticoids to induce remission followed by methotrexate for remission maintenance. METHODS: We performed an open-label prospective study in 42 patients with active Wegener's granulomatosis. All patients were treated with a standardized regimen. Outcomes were assessed using predetermined definitions based on clinical characteristics and pathologic, laboratory, and radiographic findings. RESULTS: All patients achieved disease remission. The median time to remission was 3 months, and the median time to discontinuation of glucocorticoids was 8 months. During a median of 32 months of follow-up, 1 patient died (of a myocardial infarction not related to vasculitis). Two patients (5%) had to withdraw from the study because of medication toxicity. Twenty-two patients (52%) relapsed, with glomerulonephritis occurring in 16 patients. Of these 16 patients, 4 had an increase of >0.2 mg/dL in serum creatinine level. All 4 patients returned to their prior level of renal function with treatment. None of the 22 relapses met the criteria for severe disease. CONCLUSION: The use of cyclophosphamide and glucocorticoids for induction and methotrexate for maintaining remission is an effective and well tolerated therapeutic approach in patients with active Wegener's granulomatosis. PMID- 12727580 TI - Effect of genistein on endothelial function in postmenopausal women: a randomized, double-blind, controlled study. AB - PURPOSE: Genistein, a phytoestrogen found in soybeans, corrects endothelial dysfunction induced by oophorectomy in animals. Using a double-blind, controlled, randomized design, we evaluated its effects on endothelial function in women. METHODS: We enrolled 79 healthy postmenopausal women (mean [+/- SD] age, 56 +/- 4 years) and randomly assigned them to receive continuous estrogen/progestin therapy (n = 26; 17beta-estradiol [1 mg/d] combined with norethisterone acetate [0.5 mg/d]), genistein (n = 27; 54 mg/d), or placebo (n = 26). Brachial artery flow-mediated, endothelium-dependent vasodilation and plasma levels of nitrites/nitrates (a marker of nitric oxide metabolism) and endothelin-1 were measured at baseline and after 1 year of therapy. RESULTS: Treatment with genistein increased levels of nitrites/nitrates (mean increase, 21 micromol/L; 95% confidence interval [CI]: 15 to 26 micromol/L; P <0.001 vs. placebo); estrogen/progestin therapy caused similar changes (P <0.001 vs. placebo). Plasma endothelin-1 levels decreased following 12 months of genistein (mean decrease, 7 pg/mL; 95% CI: 3 to 10 pg/mL; P <0.001 vs. placebo) and after 12 months of estrogen/progestin (P <0.001 vs. placebo). When compared with placebo, brachial artery flow-mediated dilation was improved by genistein (mean increase, 5.5%; 95% CI: 3.9% to 7.0%; P <0.001) and by estrogen/progestin (P <0.001). There were no significant differences between estrogen and genistein for any of these parameters (all P >0.4). CONCLUSION: One year of genistein therapy improves endothelium function in postmenopausal women to a similar extent as does an estrogen/progestin regimen. PMID- 12727581 TI - Potential cost-effectiveness of C-reactive protein screening followed by targeted statin therapy for the primary prevention of cardiovascular disease among patients without overt hyperlipidemia. AB - BACKGROUND: Evidence suggests that statin therapy reduces the rate of cardiovascular events among patients with low lipid levels but elevated C reactive protein levels. However, no cost-effectiveness analyses have been performed to assist in determining whether large-scale randomized trials are merited to test this hypothesis. METHODS: We used a Markov model to estimate the benefits, costs, and incremental cost-effectiveness of C-reactive protein screening followed by targeted statin therapy for elevated C-reactive protein levels, compared with dietary counseling alone, for the primary prevention of cardiovascular events among patients with low-density lipoprotein cholesterol levels <149 mg/dL. All costs were in 2000 U.S. dollars. RESULTS: The potential incremental cost-effectiveness ratio for screening followed by statin therapy compared with no screening and no statin therapy was $48,100 per quality-adjusted life-year (QALY) for 58-year-old men and $94,400 per QALY for 58-year-old women. Screening was most cost-effective for 65-year-old men ($42,600 per QALY) and least cost-effective for 35-year-old women ($207,300 per QALY). Our results were most sensitive to the baseline risk of coronary heart disease, the cost of statin therapy, and the efficacy of statin therapy for preventing myocardial infarction in patients with high C-reactive protein levels. If a 58-year-old man who smokes and is hypertensive was considered, screening for C-reactive protein followed by statin therapy would be cost saving if the cost of statin therapy was reduced to $500 per year. If the cost of statin therapy was reduced to $1 per day, the cost effectiveness of screening would be $4900 per QALY for 58-year-old men and $19,600 per QALY for women of the same age. If the costs associated with elective revascularization (percutaneous coronary intervention or coronary artery bypass surgery) were included in the base case analyses, the incremental cost effectiveness ratios for screening would be $40,100 per QALY for 58-year-old men and $87,300 per QALY for women. CONCLUSION: A strategy involving C-reactive protein screening to target statin therapy for the primary prevention of cardiovascular disease among middle-aged patients without overt hyperlipidemia could be relatively cost-effective and, in some cases, cost saving. PMID- 12727582 TI - Familial hyperkalemic hypertension: phenotypic analysis in a large family with the WNK1 deletion mutation. PMID- 12727583 TI - Importance of elevated jugular venous pressure and a third heart sound in asymptomatic left ventricular dysfunction. PMID- 12727584 TI - Beyond type 2 diabetes: the need for a clinically useful way to identify insulin resistance. PMID- 12727585 TI - Translation of basic research into useful treatments: how often does it occur? PMID- 12727586 TI - Cases from the Osler Medical Service at Johns Hopkins University. Antiglomerular basement membrane disease. AB - A 47-year-old Taiwanese man with no notable medical history was admitted with low grade fevers and night sweats that had persisted for 5 to 6 weeks. An extensive investigation at another hospital could not determine the cause of the fevers, but documented acute renal failure with a blood urea nitrogen level of 60 mg/dL and a serum creatinine level of 5.6 mg/dL. He was admitted to the Johns Hopkins Hospital for further evaluation.The patient, who had been living in the United States for the past 20 years, reported no recent travel and no behaviors that are associated with transmission of human immunodeficiency virus. He was not taking any medications, and he denied using herbal or nutritional supplements. He had no recent weight loss. There were no specific complaints on review of systems. On physical examination, he was a thin, middle-aged man in no distress. Vital signs included a temperature of 37.5 degrees C, a blood pressure of 166/86 mm Hg, a pulse of 70 beats per minute, a respiratory rate of 16 breaths per minute, and 99% oxygen saturation on room air. Sclera were anicteric, and he had no palpable adenopathy. His lungs were clear, and his heart rate was regular without extra sounds. His abdomen was thin, nontender, and without masses or organomegaly. There was no edema or signs of embolism in the extremities. Laboratory studies revealed a white blood cell count of 14,200/mL(3), a hematocrit of 23.1%, and a platelet count of 456,000/mL(3). Blood chemistries were notable for a blood urea nitrogen level of 61 mg/dL and a serum creatinine level of 7.6 mg/dL. Levels of aminotransferases, total bilirubin, and alkaline phosphatase were within normal limits. Urinalysis revealed large hemoglobin, 1+ protein, numerous red blood cells, and 3 to 5 white blood cells. Numerous red blood cell casts were seen on microscopic examination of the urine sediment. The patient's erythrocyte sedimentation rate was >130 mm/h, and his C-reactive protein level was elevated at 12.6 mg/dL. Serologies were negative for antinuclear antibodies and antineutrophil cytoplasmic antibodies; serum complement levels were normal. What is the diagnosis? PMID- 12727587 TI - Epstein-Barr virus in concomitant gastric carcinoma and adult T-cell leukemia/lymphoma. PMID- 12727588 TI - Geography of randomized controlled trials in general internal medicine: is the United States' share declining? PMID- 12727589 TI - Grapefruit and tonic: a deadly combination in a patient with the long QT syndrome. PMID- 12727590 TI - The well-being of physicians. PMID- 12727591 TI - Teaching sustainability. PMID- 12727592 TI - Converging paradigms for environmental health theory and practice. AB - Converging themes from the fields of environmental health, ecology and health, and human ecology highlight opportunities for innovation and advancement in environmental health theory and practice. In this commentary we outline the role of research and applied programs that integrate biophysical and social sciences with environmental health practice in order to address deficiencies in each field when taken on its own. New opportunities for environmental health protection and promotion are outlined based on the three converging themes: integrated approaches to research and policy, methodological acknowledgment of the synergies between the social and biophysical environments, and incorporation of core ecosystem principles into research and practice. These converging themes are discussed in relation to their implications for new types of intervention to achieve health gains across different spatial and temporal scales at the interface between biophysical and social environments. PMID- 12727594 TI - Meta-analysis of dioxin cancer dose response for three occupational cohorts. AB - This article presents a meta analysis of data from three cohorts occupationally exposed to TCDD and related compounds. A statistically significant (p = 0.02) trend was found in total cancer mortality with increasing dioxin exposure. The trend tests show an increase in total cancer at cumulative TEQ (unit of measurement for TCDD-like compounds that is defined as the amount of TCDD that would produce the same toxicity as a mixture of TCDD-like compounds) serum levels that would result from lifetime intake of 7 pg TEQ/kg body weight/day, with no increase at 6 pg/kg/day. A linear dose response provided a good fit to the combined data and predicted an ED(01) (dioxin exposure resulting in a 0.01 increase in lifetime risk of cancer mortality) of 45 pg/kg/day (95% confidence interval, 21-324). The U.S. Environmental Protection Agency estimates that current lifetime human exposures to dioxin average approximately 1 pg/kg/day (99% percentile: 3 pg/kg/day). Although it appears unlikely that current exposures through foods would reach either 7 pg/kg/day or the ED(01), our analysis argues for careful consideration of the upper ranges of long-term average exposures for dioxins. PMID- 12727593 TI - Association of expired nitric oxide with occupational particulate exposure. AB - Particulate air pollution has been associated with adverse respiratory health effects. This study assessed the utility of expired nitric oxide to detect acute airway responses to metal-containing fine particulates. Using a repeated-measures study design, we investigated the association between the fractional concentration of expired nitric oxide (F(E)NO) and exposure to particulate matter with an aerodynamic mass median diameter of less than or equal to 2.5 micro m (PM(2.5)) in boilermakers exposed to residual oil fly ash and metal fumes. Subjects were monitored for 5 days during boiler repair overhauls in 1999 (n = 20) or 2000 (n = 14). The Wilcoxon median baseline F(E)NO was 10.6 ppb [95% confidence interval (CI): 9.1, 12.7] in 1999 and 7.4 ppb (95% CI: 6.7, 8.0) in 2000. The Wilcoxon median PM(2.5) 8-hr time-weighted average was 0.56 mg/m(3) (95% CI: 0.37, 0.93) in 1999 and 0.86 mg/m(3) (95% CI: 0.65, 1.07) in 2000. F(E)NO levels during the work week were significantly lower than baseline F(E)NO in 1999 (p < 0.001). A significant inverse exposure-response relationship between log-transformed F(E)NO and the previous workday's PM(2.5) concentration was found in 1999, after adjusting for smoking status, age, and sampling year. With each 1 mg/m(3) incremental increase in PM(2.5) exposure, log F(E)NO decreased by 0.24 (95% CI: -0.38, -0.10) in 1999. The lack of an exposure-response relationship between PM(2.5) exposure and F(E)NO in 2000 could be attributable to exposure misclassification resulting from the use of respirators. In conclusion, occupational exposure to metal-containing fine particulates was associated with significant decreases in F(E)NO in a survey of workers with limited respirator usage. PMID- 12727595 TI - The relationship between water concentrations and individual uptake of chloroform: a simulation study. AB - We simulated the relationship between water chloroform concentrations and chloroform uptake in pregnant women to assess the potential extent of exposure measurement error in epidemiologic studies of the health effects of exposure to water disinfection by-products. Data from the literature were used to assign statistical distributions to swimming pool chloroform concentrations, frequency and duration of swimming, showering and bathing, and average tap water consumption. Measured increases in blood chloroform concentrations after these activities were used to estimate average uptake per microgram per liter chloroform in the water, per minute spent in the activity or per liter consumed. Given average tap water chloroform concentrations from a U.K. epidemiologic study, an average daily uptake over 90 days was simulated for 300,000 mothers. The correlation between simulated uptakes and home tap chloroform concentration was 0.6. Mothers who swam regularly received far greater doses than did nonswimmers. Results suggest there will be considerable attenuation in risk estimates and/or power loss in epidemiologic studies if the putative agent is chloroform. PMID- 12727596 TI - Disturbed sexual characteristics in male mosquitofish (Gambusia holbrooki) from a lake contaminated with endocrine disruptors. AB - Previous laboratory studies have demonstrated that estrogenic and antiandrogenic chemicals can alter several sexual characteristics in male poeciliid fishes. Whether similar disturbances occur under field conditions remains to be confirmed. Lake Apopka, Florida, is contaminated with numerous chemicals, some of which possess endocrine-disrupting activity. Male mosquitofish (Gambusia holbrooki) were collected monthly from December 2000 through May 2001 from Lake Apopka and two nearby reference lakes, Orange Lake and Lake Woodruff National Wildlife Refuge. Selected sexual characteristics were compared temporally and among lakes during the collection period. Male fish from Lake Apopka had slightly shorter gonopodia and on average 32 and 47% fewer sperm cells per milligram testis, when compared with the fish collected from Orange Lake and Lake Woodruff, respectively. The testes weights increased markedly during spring, with significantly smaller testes in fish from Lake Apopka than from Orange Lake, but surprisingly, the smallest testes occurred in males obtained from the Lake Woodruff population. The highest liver weights were found in the Lake Apopka population. Whole-body concentrations of testosterone and estradiol varied among months; the peak testosterone concentration occurred in January and was significantly lower in male fish from Lake Apopka compared with Orange Lake. The intensity of male courtship behavior was highly correlated to body testosterone concentration, but no statistically significant differences in sexual behavior among the lakes were found. We conclude that sexual characteristics of relevance to male reproductive capacity are altered in the Lake Apopka mosquitofish population, and we discuss the presence of chemicals with antiandrogenic effects in Lake Apopka as a possible cause of the observed alterations. PMID- 12727597 TI - A longitudinal examination of factors related to changes in serum polychlorinated biphenyl levels. AB - Consumption of sport-caught fish from the Great Lakes is a recognized source of human exposure to polychlorinated biphenyls (PCBs). Understanding temporal changes in PCB body burden is crucial for evaluating exposure levels and augmenting validity of studies investigating their relationship to adverse health effects. Using data collected from 1980 to 1995, we evaluated longitudinal changes in serum PCB levels among 179 fisheaters and non-fisheaters of the Michigan Fisheater Cohort. Participants identified as fisheaters in 1980 ate 26 lb or more of sport-caught fish per year, whereas non-fisheaters ate less than 6 lb per year. We found a monotonic decline in serum PCB levels among all participants from a mean value of 24 ppb in 1980 to 12 ppb in 1994. This was paralleled by an 83% decrease in mean fish consumption among all participants over the same period. We combined demographic, lifestyle, and fish consumption information with PCB data and evaluated the data using regression models to identify predictors of PCB body burden over a 16-year period. Results of the mixed-effects linear regression model suggest that consumption of Lake Michigan fish before 1980, amount of sport-caught fish eaten in the past year, age, and year of data collection were significant determinants of current PCB body burden over the 16-year study period. PCB levels were particularly elevated for males who were classified as fisheaters in 1980, which may reflect higher levels of sport-caught fish consumption compared with female fisheaters. PMID- 12727599 TI - Chronic exposure to high levels of particulate air pollution and small airway remodeling. AB - Recent evidence suggests that chronic exposure to high levels of ambient particulate matter (PM) is associated with decreased pulmonary function and the development of chronic airflow obstruction. To investigate the possible role of PM-induced abnormalities in the small airways in these functional changes, we examined histologic sections from the lungs of 20 women from Mexico City, a high PM locale. All subjects were lifelong residents of Mexico City, were never smokers, never had occupational dust exposure, and never used biomass fuel for cooking. Twenty never-smoking, non-dust-exposed subjects from Vancouver, British Columbia, Canada, a low PM region, were used as a control. By light microscopy, abnormal small airways with fibrotic walls and excess muscle, many containing visible dust, were present in the Mexico City lungs. Formal grading analysis confirmed the presence of significantly greater amounts of fibrous tissue and muscle in the walls of the airways in the Mexico City compared with the Vancouver lungs. Electron microscopic particle burden measurements on four cases from Mexico City showed that carbonaceous aggregates of ultrafine particles, aggregates likely to be combustion products, were present in the airway mucosa. We conclude that PM penetrates into and is retained in the walls of small airways, and that, even in nonsmokers, long-term exposure to high levels of ambient particulate pollutants is associated with small airway remodeling. This process may produce chronic airflow obstruction. PMID- 12727598 TI - Environmentally relevant metal and transition metal ions enhance Fc epsilon RI mediated mast cell activation. AB - Upon contact with allergen, sensitized mast cells release highly active proinflammatory mediators. Allergen-mediated mast cell activation is an important mechanism in the pathogenesis of atopic asthma. Asthmatic patients are especially susceptible to air pollution. Epidemiologic studies found a positive correlation between severity of symptoms among asthmatic patients and the level of particulate matter (PM) in the air. Among the constituents of PM are metals and transition metals, which could mediate some of its adverse effects on human health. We sought to determine the effect of metal and transition metal ions on allergen-mediated mast cell activation. We observed that several metal and transition metal ions activated mast cells and enhanced allergen-mediated mast cell activation. Thus, Al(3+), Cd(2+), and Sr(2+) induced release of granule associated N-acetyl-ss-d-hexosaminidase, and Al(3+) and Ni(2+) enhanced antigen mediated release. Metal and transition metal ions also induced significant secretion of interleukin (IL)-4 and increased antigen-mediated IL-4 secretion in mast cells. These effects of metal and transition metal ions on mast cells were observed at concentrations that do not result in direct cytotoxicity and might be relevant for environmental exposure. Thus, metals and transition metals could increase the level of allergen-mediated mast cell activation, which might be one of the mechanisms mediating exacerbation of allergen-driven asthma symptoms by air pollution. PMID- 12727600 TI - Mercury derived from dental amalgams and neuropsychologic function. AB - There is widespread concern regarding the safety of silver-mercury amalgam dental restorations, yet little evidence to support their harm or safety. We examined whether mercury dental amalgams are adversely associated with cognitive functioning in a cross-sectional sample of healthy working adults. We studied 550 adults, 30-49 years of age, who were not occupationally exposed to mercury. Participants were representative of employees at a major urban medical center. Each participant underwent a neuropsychologic test battery, a structured questionnaire, a modified dental examination, and collection of blood and urine samples. Mercury exposure was assessed using a) urinary mercury concentration (UHg); b) the total number of amalgam surfaces; and c) the number of occlusal amalgam surfaces. Linear regression analysis was used to estimate associations between each marker of mercury exposure and each neuropsychologic test, adjusting for potential confounding variables. Exposure levels were relatively low. The mean UHg was 1.7 micro g/g creatinine (range, 0.09-17.8); the mean total number of amalgam surfaces was 10.6 (range, 0-46) and the mean number of occlusal amalgam surfaces was 6.1 (range, 0-19). No measure of exposure was significantly associated with the scores on any neuropsychologic test in analyses that adjusted for the sampling design and other covariates. In a sample of healthy working adults, mercury exposure derived from dental amalgam restorations was not associated with any detectable deficits in cognitive or fine motor functioning. PMID- 12727601 TI - Health impacts of pesticide exposure in a cohort of outdoor workers. AB - We compared mortality of 1,999 outdoor staff working as part of an insecticide application program during 1935-1996 with that of 1,984 outdoor workers not occupationally exposed to insecticides, and with the Australian population. Surviving subjects also completed a morbidity questionnaire. Mortality was significantly higher in both exposed and control subjects compared with the Australian population. The major cause was mortality from smoking-related diseases. Mortality was also significantly increased in exposed subjects for a number of conditions that do not appear to be the result of smoking patterns. Compared with the general Australian population, mortality over the total study period was increased for asthma [standardized mortality ratio (SMR) = 3.45; 95% confidence interval (CI), 1.39-7.10] and for diabetes (SMR = 3.57; 95% CI, 1.16 8.32 for subjects working < 5 years). Mortality from pancreatic cancer was more frequent in subjects exposed to 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (SMR = 5.27; 95% CI, 1.09-15.40 for subjects working < 3 years). Compared with the control population, mortality from leukemia was increased in subjects working with more modern chemicals (standardized incidence ratio = 20.90; 95% CI, 1.54 284.41 for myeloid leukemia in the highest exposure group). There was also an increase in self-reported chronic illness and asthma, and lower neuropsychologic functioning scores among surviving exposed subjects when compared with controls. Diabetes was reported more commonly by subjects reporting occupational use of herbicides. These findings lend weight to other studies suggesting an association between adverse health effects and exposure to pesticides. PMID- 12727602 TI - Recruitment, retention, and compliance results from a probability study of children's environmental health in economically disadvantaged neighborhoods. AB - The School Health Initiative: Environment, Learning, and Disease (SHIELD) study used a probability sample of children (second through fifth grades) from two low income and racially mixed neighborhoods of Minneapolis, Minnesota, to assess childhood environmental health. Children were eligible to participate in SHIELD regardless of whether they or their families spoke a foreign language, their household had a telephone, or they were enrolled in a special education program. The overall enrollment rate in year 1 was 57%, with a substantial disparity between children from English-speaking (42%) versus non-English-speaking (71%) families. At the end of year 1, 85% were retained in the study. A relatively high percentage of children provided the two requested blood (82%) and urine (86%) samples in year 1, and 90% provided a valid spirometry sample. Eighty-two percent provided both requested volatile organic chemical badge samples, and both time activity logs were obtained from 66%. However, only 32% provided both peak flow measurements. All percentages increased for those participating in the second year of the study. Results indicate that a school-based research design makes it feasible and practical to conduct probability-based assessments of children's environmental health in economically disadvantaged and ethnically diverse neighborhoods. There is an ongoing need, however, to improve understanding of the cultural, economic, psychologic, and social determinants of study participation among this population. PMID- 12727603 TI - Blood lead levels and sexual maturation in U.S. girls: the Third National Health and Nutrition Examination Survey, 1988-1994. AB - Using data from the Third National Health and Nutrition Examination Survey, we assessed measures of puberty in U.S. girls in relation to blood lead levels to determine whether sexual maturation may be affected by current environmental lead exposure. The study sample included 1,706 girls 8-16 years old with pubic hair and breast development information; 1,235 girls 10-16 years old supplied information on menarche. Blood lead concentrations (range = 0.7-21.7 micro g/dL) were categorized into three levels: 0.7-2.0, 2.1-4.9, and 5.0-21.7 micro g/dL. Sexual maturation markers included self-reported attainment of menarche and physician determined Tanner stage 2 pubic hair and breast development. Girls who had not reached menarche or stage 2 pubic hair had higher blood lead levels than did girls who had. For example, among girls in the three levels of blood lead described above, the unweighted percentages of 10-year-olds who had attained Tanner stage 2 pubic hair were 60.0, 51.2, and 44.4%, respectively, and for girls 12 years old who reported reaching menarche, the values were 68.0, 44.3, and 38.5%, respectively. The negative relation of blood lead levels with attainment of menarche or stage 2 pubic hair remained significant in logistic regression even after adjustment for race/ethnicity, age, family size, residence in metropolitan area, poverty income ratio, and body mass index. In conclusion, higher blood lead levels were significantly associated with delayed attainment of menarche and pubic hair among U.S. girls, but not with breast development. PMID- 12727605 TI - Contemporary-use pesticides in personal air samples during pregnancy and blood samples at delivery among urban minority mothers and newborns. AB - We have measured 29 pesticides in plasma samples collected at birth between 1998 and 2001 from 230 mother and newborn pairs enrolled in the Columbia Center for Children's Environmental Health prospective cohort study. Our prior research has shown widespread pesticide use during pregnancy among this urban minority cohort from New York City. We also measured eight pesticides in 48-hr personal air samples collected from the mothers during pregnancy. The following seven pesticides were detected in 48-83% of plasma samples (range, 1-270 pg/g): the organophosphates chlorpyrifos and diazinon, the carbamates bendiocarb and 2 isopropoxyphenol (metabolite of propoxur), and the fungicides dicloran, phthalimide (metabolite of folpet and captan), and tetrahydrophthalimide (metabolite of captan and captafol). Maternal and cord plasma levels were similar and, except for phthalimide, were highly correlated (p < 0.001). Chlorpyrifos, diazinon, and propoxur were detected in 100% of personal air samples (range, 0.7 6,010 ng/m(3)). Diazinon and propoxur levels were significantly higher in the personal air of women reporting use of an exterminator, can sprays, and/or pest bombs during pregnancy compared with women reporting no pesticide use or use of lower toxicity methods only. A significant correlation was seen between personal air level of chlorpyrifos, diazinon, and propoxur and levels of these insecticides or their metabolites in plasma samples (maternal and/or cord, p < 0.05). The fungicide ortho-phenylphenol was also detected in 100% of air samples but was not measured in plasma. The remaining 22 pesticides were detected in 0 45% of air or plasma samples. Chlorpyrifos, diazinon, propoxur, and bendiocarb levels in air and/or plasma decreased significantly between 1998 and 2001. Findings indicate that pesticide exposures are frequent but decreasing and that the pesticides are readily transferred to the developing fetus during pregnancy. PMID- 12727604 TI - Septic system density and infectious diarrhea in a defined population of children. AB - One-quarter of U.S. households use a septic system for wastewater disposal. In this study we investigated whether septic system density was associated with endemic diarrheal illness in children. Cases--children 1 to < 19 years old seeking medical care for acute diarrhea--and controls resided in the Marshfield Epidemiologic Study Area, a population-based cohort in central Wisconsin. Enrollment was from February 1997 through September 1998. Study participants completed a structured interview, and septic system density was determined from county sanitary permits. Household wells were sampled for bacterial pathogens and indicators of water sanitary quality. Risk factors were assessed for cases grouped by diarrhea etiology. In multivariate analyses, viral diarrhea was associated with the number of holding tank septic systems in the 640-acre section surrounding the case residence [adjusted odds ratio (AOR), 1.08; 95% confidence interval (CI), 1.02-1.15; p = 0.008], and bacterial diarrhea was associated with the number of holding tanks per 40-acre quarter-quarter section (AOR, 1.22; 95% CI, 1.02-1.46; p = 0.026). Diarrhea of unknown etiology was independently associated with drinking from a household well contaminated with fecal enterococci (AOR, 6.18; 95% CI, 1.22-31.46; p = 0.028). Septic system densities were associated with endemic diarrheal illness in central Wisconsin. The association should be investigated in other regions, and standards for septic systems should be evaluated to ensure that the public health is protected. PMID- 12727606 TI - Cradle-to-cradle stewardship of drugs for minimizing their environmental disposition while promoting human health. I. Rationale for and avenues toward a green pharmacy. AB - Since the 1980s, the occurrence of pharmaceuticals and personal care products (PPCPs) as trace environmental pollutants, originating primarily from consumer use and actions rather than manufacturer effluents, continues to become more firmly established. Although PPCPs typically have been identified in surface and ground waters, some are also undoubtedly associated with solid phases such as suspended particulates, sediments, and sewage sludges, despite their relatively high affinity for water. Often amenable to degradation, their continual introduction to waste-receiving waters results from their widespread, continuous, combined use by individuals and domestic animals, giving PPCPs a "pseudo persistence" in the environment. Little is known about the environmental or human health hazards that might be posed by chronic, subtherapeutic levels of these bioactive substances or their transformation products. The continually growing, worldwide importance of freshwater resources, however, underscores the need for ensuring that any aggregate or cumulative impacts on (or from) water supplies are minimized. Despite the paucity of effects data from long-term, simultaneous exposure at low doses to multiple xenobiotics (particularly non-target-organism exposure to PPCPs), a wide range of proactive actions could be implemented to reduce or minimize the introduction of PPCPs to the environment. Most of these actions fall under what could be envisioned as a holistic stewardship program- overseen by the health care industry and consumers alike. Significantly, such a stewardship program would benefit not just the environment; additional, collateral benefits could automatically accrue, including reducing consumers' medication expenses and improving patient health and consumer safety. In this article, the first of a two-part mini-monograph describing the "green pharmacy," I focus initially on the background behind the imperative for an ecologically oriented stewardship program for PPCPs. I then present a broad spectrum of possible source control/reduction actions, controlled largely by the health care industry, that could minimize the disposition of PPCPs to the environment. This two-part mini-monograph attempts to capture cohesively for the first time the wide spectrum of actions available for minimizing the release of PPCPs to the environment. A major objective is to generate an active dialog or debate across the many disciplines that must become actively involved to design and implement a successful approach to life-cycle stewardship of PPCPs. PMID- 12727607 TI - Cradle-to-cradle stewardship of drugs for minimizing their environmental disposition while promoting human health. II. Drug disposal, waste reduction, and future directions. AB - Since the 1980s, the occurrence of pharmaceuticals and personal care products (PPCPs) as trace environmental pollutants, originating primarily from consumer use and actions rather than manufacturer effluents, continues to become more firmly established. The growing, worldwide importance of freshwater resources underscores the need for ensuring that any aggregate or cumulative impacts on (or from) water supplies are minimized. Despite a paucity of effects data from long term, simultaneous exposure at low doses to multiple xenobiotics (particularly non-target-organism exposure to PPCPs), a wide range of proactive actions could be implemented for reducing or minimizing the introduction of PPCPs to the environment. Most of these actions fall under what could be envisioned as a holistic stewardship program--overseen by the health care industry and consumers alike. Significantly, such a stewardship program would benefit not just the environment--additional, collateral benefits could automatically accrue, including the lessening of medication expense for the consumer and improving patient health and consumer safety. In this article (the second of two parts describing the "green pharmacy") I focus on those actions and activities tied more closely to the end user (e.g., the patient) and issues associated with drug disposal/recycling that could prove useful in minimizing the environmental disposition of PPCPs. I also outline some recommendations and suggestions for further research and pose some considerations regarding the future. In this mini monograph I attempt to capture cohesively for the first time the wide spectrum of actions available for minimizing the release of PPCPs to the environment. A major objective is to generate an active dialog or debate across the many disciplines that must become actively involved to design and implement a successful approach to life-cycle stewardship of PPCPs. PMID- 12727608 TI - Framework convention on tobacco control: progress and implications for health and the environment. PMID- 12727609 TI - Carcinogenicity of EBDCs. PMID- 12727611 TI - Comment on "Use of A-bomb survivor studies as a basis for nuclear worker compensation". PMID- 12727612 TI - Re: "Use of A-bomb survivor studies as a basis for nuclear worker compensation". PMID- 12727614 TI - Short-circuiting environmental protections? PMID- 12727615 TI - Tobacco's profit, workers' loss? PMID- 12727616 TI - Organic electronics: a cleaner substitute for silicon. PMID- 12727618 TI - Activation-induced cytidine deaminase links class switch recombination and somatic hypermutation. AB - Activation-induced cytidine deaminase (AID), a putative RNA-editing cytidine deaiminase that is expressed strictly in activated B cells, is indispensable in three apparently distinct genetic alterations of immunoglobulin genes-namely, class switch recombination, somatic hypermutation, and gene conversion. Recent findings led us to propose a common DNA cleaving mechanism, in which the transient secondary structure of the S and V region DNA is recognized by a nicking enzyme regulated by the putative RNA-editing activity of AID. PMID- 12727619 TI - Creation versus destruction of T cell epitopes in the class II MHC pathway. AB - Proteases perform two key roles in the class II MHC antigen processing pathway. They initiate removal of the invariant chain chaperone for class II MHC and they generate peptides from foreign and self proteins for eventual capture and display to T cells. How a balance is achieved between generation of suitable peptides versus their complete destruction in an aggressive proteolytic environment is not known. Nor is it known in most cases which proteases are actually involved in antigen processing. Our recent studies have identified asparagine endopeptidase (AEP or legumain) as an enzyme that contributes to both productive and destructive antigen processing in the class II MHC pathway. The emerging consensus seems to be that individual proteolytic enzymes make clear and non redundant contributions to antigen processing. PMID- 12727620 TI - Dendritic cell function in vivo during the steady state: a role in peripheral tolerance. AB - The avoidance of autoimmunity requires mechanisms to actively silence or tolerize self reactive T cells in the periphery. During infection, dendritic cells are not only capturing microbial antigens, but also are processing self antigens from dying cells as well as innocuous environmental proteins. Since the dendritic cells are maturing in response to microbial and other stimuli, peptides will be presented from both noxious and innocuous antigens. Therefore it would be valuable to have mechanisms whereby dendritic cells, prior to infection, establish tolerance to those self and environmental antigens that can be processed upon pathogen encounter. In the steady state, prior to acute infection and inflammation, dendritic cells are in an immature state and not fully differentiated to carry out their known roles as inducers of immunity. These immature cells are not inactive, however. They continuously circulate through tissues and into lymphoid organs, capturing self antigens as well as innocuous environmental proteins. Recent experiments have provided direct evidence that antigen-loaded immature dendritic in vivo silence T cells either by deleting them or by expanding regulatory T cells. In this way, it is proposed that the immune system overcomes at least some of the risk of developing autoimmunity and chronic inflammation. It is proposed that dendritic cells play a major role in defining immunologic self, not only centrally in the thymus but also in the periphery. PMID- 12727621 TI - Molecular mechanisms of B cell antigen receptor trafficking. AB - B lymphocytes are among the most efficient cells of the immune system in capturing, processing, and presenting MHC class II restricted peptides to T cells. Antigen capture is essentially restricted by the specificity of the clonotypic antigen receptor expressed on each B lymphocyte. However, receptor recognition is only one factor determining whether an antigen is processed and presented. The context of antigen encounter is crucial. In particular, polyvalent arrays of repetitive epitopes, indicative of infection, accelerate the delivery of antigen to specialized processing compartments, and up-regulate the surface expression of MHC class II and co-stimulatory molecules such as B7. Recent studies have demonstrated that receptor-mediated signaling and receptor facilitated peptide presentation to T cells are intimately related. For example, rapid sorting of endocytosed receptor complexes through early endosomes requires the activation of the tyrosine Syk. This proximal kinase initiates all BCR dependent signaling pathways. Subsequent entry into the antigen-processing compartment requires the tyrosine phosphorylation of the BCR constituent Igalpha and direct recruitment of the linker protein BLNK. Signals from the BCR also regulate the biophysical and biochemical properties of the targeted antigen processing compartments. These observations indicate that the activation and recruitment of signaling molecules by the BCR orchestrate a complex series of cellular responses that favor the presentation of even rare or low-affinity antigens if encountered in contexts indicative of infection. The requirement for BCR signaling provides possible mechanisms by which cognate B:T cell interactions can be controlled by the milieu in which antigen engagement occurs. PMID- 12727622 TI - Activation of rheumatoid factor (RF) B cells and somatic hypermutation outside of germinal centers in autoimmune-prone MRL/lpr mice. AB - Two critical questions need to be answered concerning the origins of autoreactive B cells in autoimmunity. First, how are autoreactive B cells regulated in normal situations? Second, how do such B cells escape tolerance mechanisms during autoimmunity? To address these questions, an Ig transgenic (Tg) mouse system based on the rheumatoid factor (RF) specificity has been developed. Tg mice express either the H or both H and L chains from AM14, an MRL/lpr-derived RF. Using this system, it was first shown that RF B cells are neither tolerized nor activated in a normal mouse. New insights into the timing and sites of initial RF B cell activation in MRL/lpr mice have been gained recently. RF B cells are activated. It was found, unexpectedly, that RF B cell activation, somatic hypermutation, and selection take place outside of the germinal center. We discuss the implications of this for the regulation of autoreactive B cells as well as for the regulation of hypermutation. PMID- 12727623 TI - Coordination of T cell activation and migration through formation of the immunological synapse. AB - T cell activation is based on interactions of T cell antigen receptors with MHC peptide complexes in a specialized cell-cell junction between the T cell and antigen-presenting cell-the immunological synapse. The immunological synapse coordinates naive T cell activation and migration by stopping T cell migration with antigen-presenting cells bearing appropriate major histocompatibility complex (MHC) peptide complexes. At the same time, the immunological synapse allows full T cell activation through sustained signaling over a period of several hours. The immunological synapse supports activation in the absence of continued T cell migration, which is required for T cell activation through serial encounters. Src and Syk family kinases are activated early in immunological synapse formation, but this signaling process returns to the basal level after 30 min; at the same time, the interactions between T cell receptors (TCRs) and MHC peptides are stabilized within the immunological synapse. The molecular pattern of the mature synapse in helper T cells is a self-stabilized structure that is correlated with cytokine production and proliferation. I propose that this molecular pattern and its specific biochemical constituents are necessary to amplify signals from the partially desensitized TCR. PMID- 12727624 TI - Intrinsic T cell defects in systemic autoimmunity. AB - Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by loss of T cell tolerance to nuclear antigens. Studies in mice and humans have demonstrated that T cells from individuals with lupus are abnormal. Here, we review the known T cell defects in lupus and their possible biochemical nature, genetic causes, and significance for lupus pathogenesis. PMID- 12727625 TI - Opsonization of apoptotic cells and its effect on macrophage and T cell immune responses. AB - Genetic studies in mice indicate that predisposition to lupus-like diseases is caused by at least three mechanisms: (1) alterations in the threshold of activation of lymphocytes or macrophages; (2) defective signaling for activation induced cell death; and (3) reduced clearance of apoptotic cells. To define the mechanisms whereby lupus develops in mice with deficiencies in either C1q, serum amyloid P component (SAP, the mouse counterpart of C-reactive protein, or CRP), or serum IgM, we studied the efficiency of phagocytosis of apoptotic cells using serum with varying levels of C1q, CRP, or IgM; we also examined the immune response to ingestion of dying cells under these conditions. Deficiency of C1q led to impaired macrophage phagocytosis of apoptotic cells, whereas CRP augmented phagocytosis, largely through recruitment of the early complement components. Like CRP, normal polyclonal IgM bound to apoptotic cells and activated complement on the cell surface. Similarly, direct binding as well as absorption experiments revealed that CRP and IgM antibodies had a similar ligand recognition specificity, namely lysophospholipids containing phosphorylcholine. IL-12 provides a pivotal link between macrophages and the T cell response to ingested material. We observed that necrotic cells induced IL-12 p40 expression, whereas phagocytosis of apoptotic cells profoundly reduced IL-12 production from stimulated macrophages. Furthermore, soluble factors from macrophages that had ingested apoptotic cells suppressed interferon-gamma production by activated T cells. These findings suggest that phospholipid exposure on apoptotic cells promotes opsonization by serum proteins leading to activation of complement, macrophage ingestion, and T cell suppression. We discuss how deficient opsonization or processing of dying cells leads to autoimmunity. PMID- 12727626 TI - Major peptide autoepitopes for nucleosome-centered T and B cell interaction in human and murine lupus. AB - The potential cross-reactivity of normal T and B cells to nuclear antigens is vast, probably due to their "education" by apoptotic cell antigens in generative lymphoid organs. Despite this "nucleocentric repertoire," as we call it, the peripheral immune system normally remains tolerant or ignorant of the products of apoptosis. However, the T helper (Th) cells, and also B cells of lupus, have a regulatory defect in the expression of CD40 ligand (CD40L). A sustained hyper expression of CD40L by lupus T cells can be triggered by sub-threshold stimuli, and is associated with impaired phosphorylation of Cbl-b, a critical downregulatory molecule in T cell signal transduction. This CD40L hyper expression abnormally prolongs co-stimulatory signals to autoimmune B cells, and it probably instigates APC (dendritic cells, resting anti-DNA B cells, and macrophages) to present apoptotic cell autoantigens in an immunogenic fashion. We have identified the dominant nucleosomal epitopes that are critical for cognate interactions between autoimmune Th cells and anti-DNA B cells in lupus. By scanning of overlapping synthetic peptides, and by mass spectrometry of naturally processed peptides, five major epitopes in nucleosomal histones were localized, namely H1'(22-42), H2B(10-33), H3(85-105), H4(16-39), and H4(71-94). The autoimmune T cells as well as B cells of lupus recognize these epitopes, and with age, autoantibodies against the peptide epitopes cross-react with nuclear autoantigens. Moreover, the peptide autoepitopes can be promiscuously presented and recognized by lupus T cells in the context of diverse MHC alleles. This cross reactivity opens up the possibility of developing "universally" tolerogenic peptides for therapy of lupus in humans despite their MHC diversity. Indeed, tolerogenic therapy with a single histone peptide epitope can halt the progression of established glomerulonephritis in lupus-prone mice by "tolerance spreading" that inactivates a broad spectrum of autoimmune T and B cells in concert. PMID- 12727627 TI - Mechanisms of autoantibody diversification to SLE-related autoantigens. AB - Systemic lupus erythematosus is a prototype of systemic autoimmunity with autoantibodies (autoAbs) to ribonucleoproteins such as Ro/La, snRNP, dsDNA, and other cellular constituents. A/J mice were used to explore the mechanism of autoAb diversification with recombinant proteins and synthetic peptides. Previous studies showed that Ro60(316-335) induced Abs to Ro60, La, and snRNP proteins. Specific Abs to determinants outside Ro60(316-335) were detected. Absorption experiments showed that Abs to La and snRNP proteins were due to the induction of anti-Ro60 Abs cross-reactive with these peptides. With snRNP proteins, SmD, SmB, and A-RNP as immunogens, specific patterns of intermolecular spreading were obtained in addition to Abs to the immunogens. With SmD-immunized mice, specific Abs to A-RNP and SmB were detected. With SmB as the immunogen, specific Abs to A RNP were detected in the majority of the mice. Only in a rare incident, specific Abs to SmD were induced. In A-RNP-immunized mice, only Abs to the 70-kD U1-RNP were seen. In all cases, Abs capable of precipitating snRNP particles were detected. Thus, the intermolecular epitope spreading is immunogen-dependent. Evidence for the presence of cross-reactive T cells to more than one autoAg was obtained. The Ag-dependent unique patterns of Ab diversification will facilitate analyses of patients' sera. These results have implications regarding the nature of the Ag-driven autoimmune process. PMID- 12727628 TI - Mechanisms inducing or controlling CD8+ T cell responses against self- or non self-antigens. AB - Cytotoxic T lymphocytes (CTLs) generally recognize antigens endogenously synthesized within the cells and presented in the form of peptides on class I molecules. However, a large body of evidence suggests that dendritic cells (DCs) have the capacity to capture and deliver exogenous antigens into the major histocompatibility complex (MHC) class I processing pathway. In this paper, we discuss this function, defined as cross-presentation, and how it is directed, particularly in inducing T cell tolerance, and how it requires special activating signals (such as CD40 ligand) to transform into a mechanism that provides either protective immunity or autoimmunity. PMID- 12727629 TI - Role of homeostatic chemokine and sphingosine-1-phosphate receptors in the organization of lymphoid tissue. AB - Chemokines regulate both homeostatic leukocyte recirculation and trafficking to sites of infection and inflammation. Apart from the well-established physiological functions, chemokines receive growing interest for their role in pathophysiological processes such as autoimmune diseases, cancer, and allograft rejection. The chemokine receptor CCR7, which is responsible for directing T cells, B cells, and dendritic cells (DCs) into secondary lymphoid organs and their precise positioning therein, has already been implicated in lymphoid organ infiltration by neoplastic cells and the localization of metastasis formation. We have shown that the differential expression of CCR7 by neoplastic cells in two entities of Hodgkin's disease (HD), classic HD (cHD) and the nodular lymphocyte predominant HD (NLPHD), may account for the differences observed in tumor cell dissemination within the affected lymph nodes. Because of the prominent role of the chemokine receptors CCR7 and CXCR5 in lymphocyte homing to secondary lymphoid organs, we hypothesized that they may also be involved in the action of FTY720, a synthetic immunosuppressant inducing lymphopenia. By using CXCR5 and CCR7 knockout mice, we have tested for a possible function of these receptors in the FTY720-induced migration of lymphocytes into Peyer's patches (PPs) and peripheral lymph nodes (PLNs). Lymphopenia is noticeably delayed in mice lacking CCR7, whereas CXCR5 knockout mice show a significant reduction of lymphocyte accumulation in secondary lymphoid organs that are infrequently present in these mice. However, FTY720-induced lymphocyte sequestration appears to be essentially independent of CCR7 and CXCR5. PMID- 12727630 TI - The human marginal zone B cell. AB - This study describes the features of the marginal zone (MZ) B cells of human tonsils and spleens and compares them with those of the follicular mantle (FM) B cells from the same tissues. The two B cell subpopulations displayed marked differences in phenotype, in response capacity to T cell-independent antigens and polyclonal B cell activators, and in presentation of antigens to T cells. FM B cells expressed surface CD5, and hence should be considered as B1 cells by current nomenclature. Fractionation of MZ B cells according to the presence or absence of surface IgD revealed the presence of two subsets. These subsets were characterized by different properties, including the presence of Ig V(H) gene mutations and the response capacity to TI-2 antigens, this latter property being associated with IgD-positive cells. Comparison of the data with those reported for mice revealed that human MZ B cells had strong analogies with both the murine MZ and B1 cells. In contrast, human B1 cells (that is, CD5-positive FM cells) were considerably different, an observation that should prompt further studies. Indeed, B cells with characteristics analogous to those of murine B1 cells were detected in small but definite proportions in the peripheral blood and tonsils. If the current distinction into B1 and B2 cells has to be maintained also for humans, it is likely that only these CD5-positive cells rather than the FM B cells should be called B1 cells. PMID- 12727631 TI - B cell immunity regulated by the protein kinase C family. AB - Protein kinase C (PKC) is a family of serine/threonine kinases which mediate essential cellular signals required for activation, proliferation, differentiation, and survival. Several PKC members are expressed in B lineage cells and activated by stimulation of the B cell receptor (BCR), thus suggesting a contribution of PKCs to the B cell-mediated immune response. To understand the individual roles of PKCs for B cell immunity, mice deficient for PKCbetaI/II (PKCbeta) or PKCdelta were analyzed. PKCbeta and PKCdelta play essential but distinctive roles in B cell immunity. In addition to its role in B cell activation and humoral immunity, PKCbeta was recently shown to control NF-kappaB activation and survival of mature B cells. PKCdelta on the other hand specifically regulates the induction of tolerance in self-reactive B cells. Thus, individual PCKs regulate B cell immunity specifically. PMID- 12727632 TI - A modified digestion-circularization PCR (DC-PCR) approach to detect hypermutation-associated DNA double-strand breaks. AB - Hypermutation of immunoglobulin variable region genes occurs in B cells during an immune response, and is vital for the development of high-affinity antibodies. The molecular mechanism of the hypermutation reaction is unknown, but it seems to correlate with the generation of locus-specific, double-strand breaks. These DNA breaks have been measured by ligation-mediated polymerase chain reaction (LM PCR), a technique that relies upon the ligation of a small linker to DNA breaks followed by the specific amplification of such breaks using combinations of locus and linker-specific primers. Here, we describe a modified version of the digestion-circularization PCR (DC-PCR) technique, which can be used to amplify and measure DNA breaks directly. PMID- 12727633 TI - Ectopic germinal center formation in rheumatoid synovitis. AB - Synovial inflammation in rheumatoid arthritis is closely related to the formation of ectopic lymphoid microstructures. In synovial tissue from some patients, one finds seemingly diffuse infiltrates; in others, T cells and B cells cluster in aggregates with interdigitating dendritic cells (DCs) but no follicular DCs (FDCs). In a third group, T cell/B cell follicles with germinal center (GC) reactions are generated. Within a given patient, aggregates and GCs are mutually exclusive and stable over time. Because antigen storage capacity, lymphoid density, and three-dimensional topography of GCs optimize immune responses, synovial GCs should play a crucial role in the breakdown of self-tolerance. We have identified factors critical for ectopic GCs, thereby transforming the synovial inflammatory process. Tissues with GCs produced 10- to 20-fold higher amounts of the chemokines CXCL13 and CCL21. CXCL13 derived from three sources, endothelial cells, synovial fibroblasts, and FDC networks. The level of CXCL13 transcripts strongly predicted GCs; however, some tissues had high levels of CXCL13 but lacked GCs. Tissue expression of LT-beta emerged as a second key factor. LT-beta protein was detected on follicular center and mantle zone B cells. Multivariate regression analysis identified CXCL13 and LT-beta as the only cytokines predicting GCs. Remarkably, LT-alpha did not contribute independently. The contribution of B cells to ectopic lymphoid organogenesis was not limited to LT-beta production. Rather, synovial tissue B cells were critical in regulating T cell activation. In adoptive transfer experiments in human synovium-SCID mouse chimeras, activation of synovium-derived CD4 T cells was strictly dependent on T cell/B cell follicles. Depletion of synovial tissue B cells abrogated T cell function, and non-B cell antigen-presenting cells could not maintain T cell stimulation. Unexpectedly, GC function in the rheumatoid lesion was also dependent on CD8 T cells. The majority of T cell receptors derived from CD8 T cells were shared between distinct GCs. Depletion of CD8 T cells disrupted synovial GCs, FDC networks disappeared, and transcription of LT-beta, IgG, and Igkappa declined. Follicle-sustaining CD8 T cells were located at the edge of or within the mantle zone. Cell-cell communication in the mantle zone, including CD8 T cells, appears to be critical for ectopic GC formation in rheumatoid synovitis. PMID- 12727635 TI - Hypermutation in human B cells in vivo and in vitro. AB - We develop our previous observation that a subpopulation of circulating memory IgM(+)IgD(+)CD27(+)B cells belongs to a separate pathway of differentiation in humans. This subpopulation, which represents 5-25% of peripheral B cells, is also present in spleen in the same proportion and displays a marginal-zone-like B cell phenotype. In addition, we describe a short-time in vitro induction model for somatic hypermutation by using the BL2 Burkitt's lymphoma cell line stimulated by a combination of antibodies directed against different surface receptors. This short-time assay allows us to show that mutations are stably introduced in one DNA strand of the BL2 VH gene in the G1 phase of the cell cycle. PMID- 12727634 TI - The V(D)J recombination/DNA repair factor artemis belongs to the metallo-beta lactamase family and constitutes a critical developmental checkpoint of the lymphoid system. AB - V(D)J recombination constitutes a critical checkpoint in the development of the immune system as shown in several animal models as well as severe combined immune deficiency (SCID) condition in humans. We recently cloned the Artemis gene, whose mutations are responsible for RS-SCID, a condition characterized by an absence of both B and T lymphocytes and associated with increased sensitivity to ionizing radiations. Artemis is ubiquitously expressed and is localized in the nucleus. Artemis belongs to the metallo-beta-lactamase superfamily and defines a new group, beta-CASP, within this family. beta-CASP proteins are beta-lactamases acting on nucleic acids. While RS-SCID patients harbor Artemis loss-of-function mutations, we identified four patients with a combined immunodeficiency characterized by a low but detectable number of both B and T lymphocytes caused by hypomorphic mutations in the Artemis gene. Two of these patients developed aggressive B cell lymphomas, a condition that suggests Artemis may be considered a "caretaker" factor, similarly to the other V(D)J recombination/DNA repair actors. PMID- 12727636 TI - Gene expression dynamics during germinal center transit in B cells. AB - The germinal center (GC) reaction in T cell dependent antibody responses is crucial for the generation of B cell memory and plays a critical role in B cell lymphomagenesis. To gain insight into the physiology of this reaction, we identified the transcriptional changes that occur in B cells during the GC transit (naive B cells --> CD77(+) centroblasts (CBs) --> CD77(-) centrocytes (CCs) --> memory B cells) by DNA microarray experiments and the subsequent data analysis employing unsupervised and supervised hierarchical clustering. The naive B cell is characterized by a nonproliferative, anti-apoptotic phenotype and the expression of various chemokine and cytokine receptors. The transition from naive B cells to CBs is associated with (1) the up-regulation of genes associated with cellular proliferation, DNA-repair, and chromatin remodeling; (2) the acquisition of a pro-apoptotic phenotype; (3) the down-regulation of cytokine, chemokine, and adhesion receptors expressed in the naive cells; and (4) the expression of a distinct adhesion repertoire. The CB and the CC revealed surprisingly few gene expression differences, suggesting that the CC is heterogeneous in its cellular composition. The CB/CC to memory B cell transition shows a general reversion to the profile characteristic for the naive B cells, with the exception of the up regulation of several surface receptors, including CD27, CD80, and IL-2Rbeta, and the simultaneous expression of both anti- and pro-apoptotic genes. These gene expression profiles of the normal B cell subpopulations are being used to identify the signals occurring during GC development, the cellular derivation of various types of B cell malignancies, and the genes deregulated in GC-derived tumors. PMID- 12727638 TI - Dendritic cells: controllers of the immune system and a new promise for immunotherapy. AB - The immune system is controlled by dendritic cells (DCs). Just as lymphocytes comprise different subsets, DCs comprise several subsets that differentially control lymphocyte function. In humans, the myeloid pathway includes Langerhans cells (LCs) and interstitial DCs (intDCs). While both subsets produce IL-12, only intDCs make IL-10 and induce B cell differentiation. Another pathway includes plasmacytoid DCs, which promptly secrete large amounts of IFN-alpha/beta viral encounter. Thus, insights into in vivo DC functions are important to understand the launching and modulation of immunity. PMID- 12727637 TI - Somatic hypermutation and B cell receptor selection in normal and transformed human B cells. AB - From the beginning to the end, the life of B cells is dominated by selection of the cells for expression of an appropriate antigen receptor. However, recent studies revealed that there are several diseases in the human where B cells lost their dependence on a B cell receptor (BCR). In classic Hodgkin's lymphoma, the lymphoma cells presumably derive from "crippled" germinal center (GC) B cells that acquired unfavorable somatic Ig gene mutations, which often render originally functional immunoglobulin (Ig) genes nonfunctional. A peculiar situation is observed among Epstein-Barr virus (EBV)-infected B cells in angioimmunoblastic lymphadenopathy with dysproteinaemia (AILD)-type T cell lymphoma, where somatic hypermutation uncoupled from any selection for functionality of the BCR is observed in expanding clones. Clones of EBV-harboring B cells that show ongoing hypermutation during proliferation and are Ig-deficient in at least a fraction of cases were recently also identified in post-transplant lymphoproliferative disorders. Hence, transformed B cells may, in particular settings, escape the normal selectional forces to express a BCR, and EBV may cause dramatic changes in B cell differentiation programs. Somatic hypermutation may be involved in lymphomagenesis by several means. Some chromosomal translocations into Ig loci likely involve DNA-strand breaks associated with hypermutation. Moreover, by aberrant targeting of the CD95 gene, GC B cells and lymphomas developing from them may become resistant to elimination by CD95 ligand expressing T cells. Finally, aberrant hypermutation of multiple proto-oncogenes appears to be a major factor in diffuse large cell lymphoma pathogenesis. PMID- 12727639 TI - Co-stimulatory blockade in the treatment of murine systemic lupus erythematosus (SLE). AB - Although the life span of patients with systemic lupus erythematosus (SLE) has improved considerably over the last several decades, the toxicities of chronic immunosuppressive therapy are major causes of morbidity and mortality. Safer and more effective therapies for SLE are clearly needed. SLE is characterized by excessive activation of both B and T lymphocytes. Activation of these cells requires both antigen engagement and co-stimulatory signals from interacting lymphocytes (Carreno, B.M. M. Collins, 2002, Annu. Rev. Immunol. 20: 29-53; Grewal, I.S. R.A. Flavell, 1998, Annu. Rev. Immunol. 16: 111-135). Thus, blockade of co-stimulatory signals offers a new therapeutic approach to SLE. Our short term goal has been to understand the effect of co-stimulatory blocking reagents on the development, selection, and activation of pathogenic anti-dsDNA antibody producing B cells in mice genetically pre-determined to develop SLE and showing signs of either early or advanced disease activity. Our long-term goal is to use the knowledge we gain to design therapeutic regimens for humans that avoid the complications of long-term immunosuppression. As new co-stimulatory molecules are discovered, studying their mechanism of action in animal models and their clinical utility in human autoimmune disease should lead both to a new understanding of disease pathogenesis and also to safer and more effective therapies. PMID- 12727640 TI - The transfer of immunity from mother to child. AB - The newborn's immune system grows fast from a small size at birth by exposure primarily to the intestinal microflora normally obtained from the mother at and after birth. While building up its immune system, the infant is supported by the transplacental IgG antibodies, which also contain anti-idiotypic antibodies, possibly also actively priming the offspring. The second mode of transfer of immunity occurs via the milk. Numerous major protective components, including secretory IgA (SIgA) antibodies and lactoferrin, are present. The breastfed infant is better protected against numerous common infections than the non breastfed. Breastfeeding also seems to actively stimulate the infant's immune system by anti-idiotypes, uptake of milk lymphocytes, cytokines, etc. Therefore, the breastfed child continues to be better protected against various infections for some years. Vaccine responses are also often enhanced in breastfed infants. Long-lasting protection against certain immunological diseases such as allergies and celiac disease is also noted. PMID- 12727641 TI - Novel method to control pathogenic bacteria on human mucous membranes. AB - Nearly all infections begin at a mucous membrane site. Also, the human mucous membranes are a reservoir for many pathogenic bacteria found in the environment (that is, pneumococci, staphylococci, streptococci), some of which are resistant to antibiotics. Clearly, if this human reservoir can be reduced or eliminated, the incidence of disease will be markedly reduced. However, compounds designed to eliminate this reservoir are not available. Towards this goal, we have exploited the highly lethal effects of bacteriophage lytic enzymes (lysins) to specifically destroy disease bacteria on mucous membranes. Such lysins are used by the phage to release their progeny at the end of their replicative cycle. We have identified and purified these enzymes and found that when applied externally to gram-positive bacteria, they are killed seconds after contact. For example, 10(7) S. pyogenes are reduced to undetectable levels 10 s after enzyme addition. A feature of these enzymes is their high specificity; that is, streptococcal lysins kill streptococci and pneumococcal lysins kill pneumococci without effects on the normal flora organisms. In vivo, an oral colonization model for S. pyogenes and a nasal colonization model for S. pneumoniae were developed to test the capacity of the lysins to kill organisms on these surfaces. In both cases, when the animals were pre-colonized with their respective bacteria then treated with a small amount of lysin, specific for the colonizing organism, all the animals were found to be free of colonizing bacteria shortly after lysin treatment. Thus, lysins may be added to our armamentarium to control antibiotic-resistant bacteria. PMID- 12727642 TI - Peptide mimics of a major lupus epitope of SmB/B'. AB - One of the most distinguishing features of systemic lupus erythematosus is the presence of high concentrations of autoantibodies that recognize a limited number of self-antigens. Even though many lupus autoantigens have been identified, the inciting triggers of these abnormal immune responses are not fully understood. One mechanism that could generate these autoantibodies is a normal immune response toward a foreign epitope that mimics a common antigenic target of an autoantigen. Antibody generated toward the foreign epitope could also bind the autoantigen. This "cross-reactivity" would result in the presentation of the autoantigen to the immune system. Under autoimmune-prone conditions, tolerance toward the native protein is broken and an autoimmune response is initiated. Previously, it was suggested that Epstein-Barr virus might use such a mechanism to initiate an autoimmune response. Cross-reactive epitopes may have a similar amino acid sequence or a similar tertiary structure that is independent of amino acid sequence. A major, and likely initial, target of the lupus anti-SmB' response is a repeated, proline-rich sequence, PPPGMRPP. To identify potential cross-reactive targets, we used affinity-purified autoantibodies specific for PPPGMRPP to screen a random heptapeptide phage display library. Eighty-five clones were isolated and sequenced with eleven distinct sequence motifs being identified. Two of these motifs were homologous to the SmB' epitope, while the other nine were not. Interestingly, one of the peptide motifs that mimicked the SmB' epitope is identical to a peptide sequence found in the Epstein-Barr virus major DNA binding protein. PMID- 12727643 TI - CD137-mediated T cell co-stimulation terminates existing autoimmune disease in SLE-prone NZB/NZW F1 mice. AB - T cell receptor recognition of antigen and major histocompatibility complex (signal 1) and T cell co-stimulation (signal 2) are essential for full T cell activation, differentiation, and survival of naive and activated T cells. The proto-typical T cell co-stimulatory receptor, CD28, is a constitutively expressed type I integral transmembrane protein and member of the Ig superfamily. Since its discovery, additional T cell co-stimulatory receptors have been identified, a number of which belong to the tumor necrosis factor receptor superfamily. Included within this group is CD137 (4-1BB), an activation-inducible, type I transmembrane protein. Co-stimulation of T cells through CD137 effectively up regulates CD8 T cell activation and survival. Although CD4(+) T cells are efficiently activated through the T cell receptor and CD137 receptor, it provokes CD4(+) T cell anergy and blockade of T-dependent humoral immune responses. Therefore, we tested whether agonistic anti-CD137 monoclonal antibodies (mAbs) would be effective in blocking the induction or progression of B cell dependent autoimmune disease. Herein, we demonstrate the protective effect of agonistic anti-CD137 mAbs in blocking systemic lupus erythematosus (SLE) disease progression in NZB/W F1 mice. Protection from SLE following anti-CD137 mAb treatment is not confined to rescuing mice from disease progression; rather, it fully protects young mice from developing any symptoms of disease. We further found that treatment of proteinuric mice with anti-CD137 blocks ongoing anti dsDNA autoantibody production. PMID- 12727644 TI - Using single-gene deletions to identify checkpoints in the progression of systemic autoimmunity. AB - Systemic lupus erythematosus is a multigenic disorder of unknown etiology. To investigate the roles that specific genes play in lupus, we have examined the disease profiles in mice with single-gene deletions. In total, some 17 genes have been studied. Absence of certain genes, such as CD40L, CD28, or Igh6, abrogated induction of autoimmunity. Other genes, such as Igh5, IL-4, or ICAM-1, had little effect on the development of disease. Intermediate effects were observed in IL-6 deficient mice, while absence of beta2-microglobulin resulted in loss of hypergammaglobulinemia and IgG1 autoantibodies, but produced little change in anti-chromatin antibodies or glomerular deposits. The most interesting observations were obtained with genes related to the expression or function of interferon-gamma (IFN-gamma). Reductions in IFN-gamma levels in murine lupus are associated with reductions in both autoantibody levels and immune-complex- mediated pathology. Genes involved in up-regulation of IFN-gamma expression, such as IL-12, STAT-4, or ICE, did not significantly influence autoimmunity, whereas absence of IFN-gamma or IFN-gamma receptor led to greatly reduced autoantibody response and immunopathology. Absence of IRF-1, a gene ex-pressed in response to IFN-gamma, resulted in selective retention of anti-chromatin antibodies but little glomerular pathology. These studies suggest that the presence of a baseline level of IFN-gamma, rather than increased expression, is important for autoimmunity. Furthermore, as the IRF-1 knockout demonstrates, specific defects in signaling pathways and gene expression subsequent to IFN-gamma/IFN-gamma receptor interaction may influence only certain disease parameters. It has not escaped our attention that IFN-gamma influences the expression and function of other immunologically relevant genes, such as IL-4, IL-6, and beta2 microglobulin. Thus, these genes may be part of the downstream events following IFN-gamma/IFN-gamma receptor interaction that promote the development of autoimmunity. PMID- 12727645 TI - Activation of the Ets transcription factor Elf-1 requires phosphorylation and glycosylation: defective expression of activated Elf-1 is involved in the decreased TCR zeta chain gene expression in patients with systemic lupus erythematosus. AB - Elf-1, a member of the Ets transcription factor family with an estimated molecular mass of 68 kDa, is involved in the transcriptional regulation of several hematopoietic cell genes. It is shown that following O-GlcNAc glycosylation and phosphorylation by PKC theta, the cytoplasm-located, 80-kDa Elf 1 translocates to the nucleus as a 98-kDa protein. In the nucleus, Elf-1 binds to the promoter of the TCR zeta gene and promotes its transcription in Jurkat and fresh human T cells. It is also shown that in the majority of patients with systemic lupus erythematosus (SLE), who are known to express decreased levels of T cell receptor (TCR) zeta chain and mRNA, the 80-kDa Elf-1 protein does not undergo proper post-transcriptional modification, which results in low levels of the 98-kDa protein, lack of Elf-binding to the TCR zeta promoter, and decreased gene transcription. Therefore, a novel activation pathway for a member of the Ets family of transcription factors, which is defective in patients with systemic autoimmunity, has been revealed. PMID- 12727646 TI - Aberrant activation of B cells in patients with rheumatoid arthritis. PMID- 12727647 TI - The role of relB in regulating the adaptive immune response. AB - Dendritic cells (DCs), which represent a key type of antigen-presenting cell (APC), are important for the development of innate and adaptive immunity. DCs are involved in T cell activation in at least two main ways: priming via direct processing/presentation of soluble antigen taken up from the microenvironment (conventional priming), and processing/presentation of antigen released from other cells (cross-priming). relB, a component of the NF-kappaB complex of transcription factors, is a critical regulator of the differentiation of DCs. In mice, lack of relB impairs DCs derived from bone marrow both in number and function. Here relB (-/-) bone marrow chimera mice is used to study the APC function of residual DCs in presentation of soluble antigen and cross-priming. It is found that the DCs in these mice are profoundly deficient in their ability to both prime and cross-prime T cell responses. It was concluded that the relB gene is involved in regulating the APC function of DCs in vivo. PMID- 12727648 TI - Tolerogenic dendritic cells induced by vitamin D receptor ligands enhance regulatory T cells inhibiting autoimmune diabetes. AB - Dendritic cells (DCs) not only induce but also modulate T cell activation. 1,25 Dihydroxyvitamin D(3) [1,25-(OH)(2)D(3)] induces DCs with a tolerogenic phenotype, characterized by decreased expression of CD40, CD80, and CD86 co stimulatory molecules, low IL-12, and enhanced IL-10 secretion. We have found that a short treatment with 1,25-(OH)(2)D(3) induces tolerance to fully mismatched mouse islet allografts, and that this tolerance is stable to challenge with donor-type spleen cells and allows acceptance of donor-type vascularized heart grafts. This effect is enhanced by co-administration of mycophenolate mofetil (MMF), a selective inhibitor of T and B cell proliferation, that also has effects similar to 1,25-(OH)(2)D(3) on DCs. Graft acceptance is associated with impaired development of type 1 CD4(+) and CD8(+) cells and an increased percentage of CD4(+)CD25(+) regulatory cells expressing CD152 in the spleen and in the draining lymph node. Transfer of CD4(+)CD25(+) cells from tolerant mice protects 100% of the syngeneic recipients from islet allograft rejection. CD4(+)CD25(+) cells that are able to inhibit the T cell response to a pancreatic autoantigen and to significantly delay disease transfer by pathogenic CD4(+)CD25( ) cells are also induced by treatment of adult nonobese diabetic (NOD) mice with a selected vitamin D receptor (VDR) ligand. This treatment arrests progression of insulitis and Th1 cell infiltration, and inhibits diabetes development at non hypercalcemic doses. The enhancement of CD4(+)CD25(+) regulatory T cells able to mediate transplantation tolerance and to arrest type 1 diabetes development by a short oral treatment with small organic compounds that induce tolerogenic DCs, like VDR ligands, suggests possible clinical applications of this approach. PMID- 12727649 TI - Antibody repertoire in a mouse with a simplified D(H) locus: the D-limited mouse. PMID- 12727650 TI - Long-lived plasma cells in immunity and inflammation. PMID- 12727651 TI - Molecular mechanism of serial VH gene replacement. AB - The molecular mechanism of serial VH replacement was analyzed using a human B cell line, EU12, that undergoes continuous spontaneous differentiation from pro-B to pre-B and then to B cell stage. In earlier studies, we found that this cell line undergoes intraclonal V(D)J diversification. Analysis of the IgH gene sequences in EU12 cells predicted the occurrence of serial VH replacement involving the cryptic recombination signal sequences (cRSS) embedded within framework 3 regions and concurrent extension of the CDR3 region. Detection of double-stranded DNA breaks at the cRSS site and different VH replacement excision circles confirm the ongoing nature of this diversification process in the EU12 cells. In vitro binding and cleavage assays using recombinant RAG-1 and RAG-2 proteins further validated the cRSS participation in this RAG-mediated recombination process. Serial VH replacements may represent an additional mechanism for diversification of the primary B cell repertoire. PMID- 12727652 TI - Oligoclonal expansions of antigen-specific CD8+ T cells in aged mice. AB - Oligoclonal T cell expansions (TCE) are common in old humans and mice. It is not known whether an Ag-specific response becomes more oligoclonal with age, and, if so, how this might alter biological responses or compromise the immune response, thus contributing to the immunodeficiency of aging. We used a tumor antigen response to study these questions. Early on, antigen reactive T cell numbers at the site of tumor injection were lower and clonally more restricted in old mice. Subsequently, long-term oligoclonal TCE emerged in the blood and spleen of old mice. IL-15 was not necessary for development of TCE in the blood. Overall, the data pointed to a dysregulated immune response in old mice, perhaps due to lack of optimal IL-2 and CD4 help at the earliest stages and a lack of an efficient local peritoneal CTL response. This was associated with a deficient humoral response and, likely, persistence of tumor cells or tumor antigens. Perhaps the spleen is the site of persistence which explains clonal TCE observed primarily in PBL and spleen. The TCE appear to be inefficient as they are often anergic. As a result an occasional peritoneal or splenic tumor may arise in old mice. PMID- 12727653 TI - A genome-wide analysis of the acute-phase response and its regulation by Stat3beta. AB - The acute-phase response (APR) is the systemic inflammatory component of innate immunity. A global assessment of hepatic gene expression during an APR has been undertaken. In response to endotoxin, an inducer of the APR, about 7% of mouse genes exhibited significant changes in expression. Genes for cholesterol, fatty acid, and phospholipid synthesis were suppressed, while genes participating in innate defense and antigen presentation were induced. Upon challenge with endotoxin, mice deficient in Stat3beta, a dominant-negative variant of Stat3, exhibited impaired recovery and susceptibility to protracted shock. These findings are accompanied by overexpression and hyperresponsiveness of a subset of lipopolysaccharide (LPS)-inducible genes in liver, suggesting a critical role for Stat3beta in the control of systemic inflammation. PMID- 12727670 TI - Invited commentary: trends in coronary heart disease mortality-location, location, location. PMID- 12727669 TI - Coronary heart disease death and sudden cardiac death: a 20-year population-based study. AB - Trends in out-of-hospital coronary heart disease (CHD) death, a surrogate for sudden cardiac death (SCD), are important for understanding the decline in CHD mortality. Little is known about out-of-hospital CHD death without prior CHD diagnosis, the definition of unexpected SCD. The authors analyzed secular trends in CHD death and unexpected SCD over a 20-year period (1979-1998) to examine the association between prior CHD and SCD and to test the hypothesis that in-hospital deaths declined more than SCDs. The yearly decline in CHD mortality rates was 5.3% for in-hospital deaths and 1.8% for out-of-hospital deaths (p = 0.001). Among all SCDs, the proportion of unexpected SCD was 49%. Mortality rates for both unexpected SCD and SCD with prior CHD declined over time, but unexpected SCD declined at a slower rate than SCD with prior CHD (p = 0.001). The relative odds of prior CHD were higher among persons with SCD than among controls, but there was a modest decline in the magnitude of the association. Thus, during the past 20 years, the decline was greater for in-hospital CHD deaths than for SCDs. Since approximately half of the SCDs were unexpected and rates of these deaths declined less over time than rates of SCD with prior CHD, primary prevention is becoming increasingly more important in sustaining the decline in CHD mortality. PMID- 12727671 TI - Effect of neighborhood social participation on individual use of hormone replacement therapy and antihypertensive medication: a multilevel analysis. AB - The authors investigated a possible contextual effect of neighborhood on individual use of hormone replacement therapy (HRT) and antihypertensive medication (AHM) and the impact of neighborhood social participation on individual use of these medications. They attempted to disentangle contextual from individual influences. Multilevel logistic regression modeling was used to analyze data on 15,456 women aged 45-73 years (first level) residing in 95 neighborhoods (second level) of the city of Malmo, Sweden (250,000 inhabitants) who participated in the Malmo Diet and Cancer Study (1991-1996). AHM use was studied among 7,558 participants with defined hypertension. Of the total variability in medication use in this population, only 1.7% (HRT) and 0.5% (AHM) was between neighborhoods. After adjustment for age, individual socioeconomic factors, individual low levels of social participation, and health and behavioral variables, no neighborhood effect on AHM use was found. However, women living in neighborhoods with low social participation were much less likely to use HRT (odds ratio = 0.36, 95% confidence interval: 0.21, 0.63), especially if they themselves experienced low social participation (synergy index, 1.53) or were immigrants (synergy index, 1.68). The Malmo neighborhoods were homogeneous with regard to HRT and especially AHM use. However, differences in neighborhood social participation affected HRT use independently of individual characteristics. PMID- 12727672 TI - Insulin-like growth factor I, insulin-like growth factor binding protein 3, and urologic measures of benign prostatic hyperplasia. AB - Laboratory studies suggest that insulin-like growth factor I (IGF-I) promotes prostatic growth. The authors evaluated the association between benign prostatic hyperplasia and IGF-I and its binding protein IGFBP-3 in community-dwelling men to determine whether this laboratory finding is manifest at the population level. Participants (n = 471) were Olmsted County, Minnesota, Caucasian males aged 40-79 years in 1990. Urologic measures were assessed from the International Prostate Symptom Score, peak urinary flow rates, prostate volume, and serum prostate specific antigen (PSA), and serum IGF-I and IGFBP-3 levels were measured. After adjustment for age, the relative odds (odds ratios) of an abnormal urologic measure in men with high versus low serum IGF-I levels were 0.98 (95% confidence interval (CI): 0.66, 1.45) for a symptom score of >7, 1.14 (95% CI: 0.72, 1.80) for a peak urinary flow rate of <12 ml/second, 1.11 (95% CI: 0.72, 1.72) for a prostate volume of >30 ml, and 0.71 (95% CI: 0.46, 1.09) for a PSA level of >1.4 ng/ml. A low IGFBP-3 level was associated with an enlarged prostate (odds ratio = 1.72, 95% CI: 1.05, 2.82), after simultaneous adjustment for IGF-I and age, but not with other urologic measures. These data do not provide evidence for an association between benign prostatic hyperplasia and serum IGF-I. PMID- 12727674 TI - Use of agricultural pesticides and prostate cancer risk in the Agricultural Health Study cohort. AB - The authors examined the relation between 45 common agricultural pesticides and prostate cancer incidence in a prospective cohort study of 55,332 male pesticide applicators from Iowa and North Carolina with no prior history of prostate cancer. Data were collected by means of self-administered questionnaires completed at enrollment (1993-1997). Cancer incidence was determined through population-based cancer registries from enrollment through December 31, 1999. A prostate cancer standardized incidence ratio was computed for the cohort. Odds ratios were computed for individual pesticides and for pesticide use patterns identified by means of factor analysis. A prostate cancer standardized incidence ratio of 1.14 (95% confidence interval: 1.05, 1.24) was observed for the Agricultural Health Study cohort. Use of chlorinated pesticides among applicators over 50 years of age and methyl bromide use were significantly associated with prostate cancer risk. Several other pesticides showed a significantly increased risk of prostate cancer among study subjects with a family history of prostate cancer but not among those with no family history. Important family history pesticide interactions were observed. PMID- 12727673 TI - Reported family history of cancer in the prostate, lung, colorectal, and ovarian cancer screening trial. AB - The authors analyzed data from almost 150,000 subjects aged 55-74 years enrolled in the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial who completed a self-administered baseline questionnaire (1993-2001) that included items about family history of cancer. Male respondents reported significantly less family history of cancer than females. The relative underreporting by male respondents relative to females was greater for female family members (28% lower for sisters and 21% lower for mothers) than for male family members (13% lower for brothers and 9% lower for fathers). Black, Hispanic, and Asian respondents reported significantly less family history of cancer than Whites. Reported family history prevalences for parents decreased with respondents' age, while those for siblings increased with respondents' age. The four most commonly reported cancers in families were breast (11.8%), lung (10.1%), colorectal (9.4%), and prostate (7.3%) cancer. Expected prevalences in family members of history of cancer overall and history of specific types of cancer were calculated using incidence rates and life table data obtained from the Surveillance, Epidemiology, and End Results Program. Overall, the ratio of reported cancer rates to expected cancer rates in family members was approximately 0.7. Liver, bone, stomach, and brain cancer had greater-than-average reported:expected ratios, while lymphoma, bladder cancer, melanoma, and testicular cancer had lower-than-average ratios. PMID- 12727675 TI - Does male age affect the risk of spontaneous abortion? An approach using semiparametric regression. AB - Couples in industrialized countries tend to delay attempting to have children, which may lower their chances of livebirth. The authors assessed the association between male age and the risk of spontaneous abortion between weeks 5 and 20 of pregnancy, controlling for female age. They interviewed by telephone a random cross-sectional population of 1,151 French women who had been pregnant between 1985 and 2000 (participation rate, 73%). A total of 12.2% of the 2,414 pregnancies resulted in spontaneous abortion. Semiparametric regression was used to define a discrete time survival model with a random effect taking into account induced abortions, in which female age was coded by a third-degree polynomial. This final model predicted that the risk (rate ratio) of spontaneous abortion was 2.13-fold higher for women age 25 years whose partner was age 35 years or older than for women age 25 years whose partner was younger than age 35 years (95% confidence interval: 1.07, 4.26). No such increased risk of spontaneous abortion with male age was estimated when the woman was age 35 years (rate ratio = 0.61, 95% confidence interval: 0.35, 1.07). Thus, increasing male age could increase the risk of spontaneous abortion when the female partner is less than 30 years of age. PMID- 12727676 TI - Injury risk in men's Canada West University football. AB - Injury and participation information was collected over 5 years (1993-1997) on varsity men's football players in the Canada West Universities Athletic Association. The locations of acute time-loss injuries or neurologic injures were coded as head and neck, upper extremity (shoulder to hand), or lower extremity (hip to foot). Poisson regression-based generalized estimating equations were used to estimate rate ratios and 95% confidence intervals. Injury rates were higher during games as compared with practice periods (for the head and neck, rate ratio (RR) = 9.75 (95% confidence interval (CI): 7.50, 12.67); for upper extremities, RR = 5.76 (95% CI: 4.46, 7.45); and for lower extremities, RR = 7.06 (95% CI: 6.03, 8.25)). In dry-field game situations, head and neck injury rates were 1.59 times higher on artificial turf than on natural grass (95% CI: 1.04, 2.42). Lower extremity game injury rates were higher on artificial turf than on natural grass under both dry (RR = 1.83, 95% CI: 1.35, 2.48) and wet (RR = 2.31, 95% CI: 1.18, 4.52) field conditions. Injury rates increased with every additional year of participation. Past injury increased the rate of subsequent injury. The effect of an artificial field surface may be related to infrequent use. Risk factors for injury included participation in a game, playing on artificial turf, being a veteran player, and having a past injury. PMID- 12727678 TI - Identifying West Nile virus risk areas: the Dynamic Continuous-Area Space-Time system. AB - The Dynamic Continuous-Area Space-Time (DYCAST) system was developed to identify and prospectively monitor high-risk areas for West Nile virus in New York, New York (New York City). The system is based on a geographic model that uses a localized Knox test to capture the nonrandom space-time interaction of dead birds, as an indicator of an intense West Nile virus amplification cycle, within a 1.5-mile (2.41-km) buffer area and 21-day moving window. The Knox analysis is implemented as an interpolation function to create a surface of probabilities over a grid of 1,400 cells overlaying New York City. The model's parameters were calibrated using year 2000 data and information on the vector-host transmission cycle. The DYCAST system was implemented in a geographic information system and used operationally in year 2001. It successfully identified areas of high risk for human West Nile virus infection in areas where five of seven human cases resided, at least 13 days prior to the onset of illness, and proved that it can be used as an effective tool for targeting remediation and control efforts. PMID- 12727677 TI - Developmental typology of trajectories to nighttime bladder control: epidemiologic application of longitudinal latent class analysis. AB - The authors aimed to characterize developmental trajectories to nighttime continence by applying two latent class models-longitudinal latent class analysis (LLCA) and latent class growth analysis (LCGA)-to data on nighttime bed-wetting from a population-based birth cohort, the Medical Research Council 1946 National Survey of Health and Development cohort. Data on a binary outcome (wetting in the past month vs. not wetting) were available for children at six ages (4, 6, 8, 9, 11, and 15 years) assessed in 1950, 1952, 1954, 1955, 1957, and 1961. For 3,272 children with complete data (62.5% of the cohort), results of sequential model comparisons (T classes vs. T + 1 classes) and chi-square goodness-of-fit tests were evaluated using parametric bootstrapping. At least four trajectory classes (LLCA and LCGA) were identified. Associations between class membership and the prevalence of related measures were examined using a confirmatory latent class analysis approach. Inclusion of 1,483 children with partially incomplete data (n = 4,755; 90.9% of the cohort) enabled the authors to refine trajectories further: normal development (prevalence = 84.0%); delayed acquisition of bladder control ("transient" (8.7%) and "persistent" (1.8%)), capturing primary enuresis; chronic bed-wetting (2.6%), or experiencing night wetting until age 15 years; and a final trajectory (relapse = 2.9%) capturing secondary or onset enuresis. This empirically based, typologic approach to analysis of extensive longitudinal data in a general population sample provides an alternative perspective to that offered by traditional diagnostic criteria. PMID- 12727679 TI - Re: "Association of transposition of the great arteries in infants with maternal exposures to herbicides and rodenticides". PMID- 12727680 TI - Clinical and biological interactions in affective and cognitive geriatric syndromes. PMID- 12727681 TI - Implications for the neural basis of social cognition for the study of schizophrenia. AB - OBJECTIVE: The study of social cognition in schizophrenia has received growing attention in recent years. At the same time, a large body of work has explored the neural basis of social cognition in both nonclinical and clinical groups, other than those with schizophrenia. The gap between these two literatures is considerable and may slow progress in creating a comprehensive social cognitive model of schizophrenia. This article attempts to bridge this gap by discussing how the neural basis of social cognition may inform future clinical research in schizophrenia. METHOD: PsycINFO and MEDLINE were systematically searched for articles pertaining to the neural basis of social cognition and social cognition in schizophrenia. Relevant studies were obtained and synthesized into a comprehensive review and integrative formulation. RESULTS: Striking parallels between these two areas of research were found. CONCLUSIONS: These parallels might help to better elucidate the underlying mechanisms for social cognitive and social behavioral impairments in schizophrenia as well as provide potential targets for treatment and drug development. PMID- 12727682 TI - Cognitive behavior therapy for depression? Choose horses for courses. AB - OBJECTIVE: Although cognitive behavior therapy is a widely accepted treatment for depression, the problematic nature of efficacy studies is insufficiently recognized. METHOD: The authors reviewed original studies and quantitative analyses on the use of cognitive behavior therapy for depression. RESULTS: The authors suggested that claims for cognitive behavior therapy's efficacy on depression have been overstated, questioned whether its efficacy fits within its theoretical underpinning, and argued against viewing cognitive behavior therapy as a universal rather than a targeted strategy. CONCLUSIONS: Although cognitive behavior therapy may act more by its nonspecific therapeutic ingredients, the authors argued that by testing cognitive behavior therapy's efficacy in heterogeneous study groups, rather than in specific subgroups, failure to differentiate it from control therapies may have been ensured. PMID- 12727683 TI - Transcranial magnetic stimulation in the treatment of depression. AB - OBJECTIVE: Transcranial magnetic stimulation (TMS) is a noninvasive and easily tolerated method of altering cortical physiology. The authors evaluate evidence from the last decade supporting a possible role for TMS in the treatment of depression and explore clinical and technical considerations that might bear on treatment success. METHOD: The authors review English-language controlled studies of nonconvulsive TMS therapy for depression that appeared in the MEDLINE database through early 2002, as well as one study that was in press in 2002 and was published in 2003. In addition, the authors discuss studies that have examined technical, methodological, and clinical treatment parameters of TMS. RESULTS: Most data support an antidepressant effect of high-frequency repetitive TMS administered to the left prefrontal cortex. The absence of psychosis, younger age, and certain brain physiologic markers might predict treatment success. Technical parameters possibly affecting treatment success include intensity and duration of treatment, but these suggestions require systematic testing. CONCLUSIONS: TMS shows promise as a novel antidepressant treatment. Systematic and large-scale studies are needed to identify patient populations most likely to benefit and treatment parameters most likely to produce success. In addition to its potential clinical role, TMS promises to provide insights into the pathophysiology of depression through research designs in which the ability of TMS to alter brain activity is coupled with functional neuroimaging. PMID- 12727684 TI - Schizophrenia, I. PMID- 12727685 TI - Scaling down. PMID- 12727686 TI - An impaired physician with complex comorbidity. PMID- 12727687 TI - Dali (1904-1989): psychoanalysis and pictorial surrealism. PMID- 12727688 TI - A collaborative study of the emergence and clinical features of the major depressive syndrome of Alzheimer's disease. AB - OBJECTIVE: This report provides a description of the prevalence and clinical features of the major depressive syndrome of Alzheimer's disease using data derived from structured diagnostic assessments of 243 patients with probable Alzheimer's disease and 151 nondemented elderly comparison subjects. METHOD: Subjects were characterized by a consortium of four Alzheimer's disease research centers and the Geriatric Psychiatry Branch of the National Institute of Mental Health. All sites administered the Clinical Assessment of Depression in Dementia, a structured, anchored diagnostic interview that was developed to reliably diagnose and characterize major depressive episodes in this population. RESULTS: Despite the use of a common, reliable methodology for the assessment and diagnosis of major depressive episodes, the prevalence of major depression in Alzheimer's disease ranged widely from 22.5% to 54.4% across the recruitment sites. The prevalence of major depressive episodes among Alzheimer's disease patients in the aggregate sample exceeded that for elderly comparison subjects and reached nearly 50% among the most severely demented patients. Alzheimer's disease patients with a current major depressive episode had earlier mean ages at onset, a higher mean Hamilton Depression Rating Scale score, and were more likely to be experiencing psychotic symptoms than those who had not developed a major depressive episode. Although the major depressive episodes of Alzheimer's disease patients and nondemented elderly comparison subjects included similar numbers of depressive symptoms, patients with Alzheimer's disease were more likely to report a diminished ability to concentrate or indecisiveness and less likely to experience sleep disturbances and feelings of worthlessness or excessive guilt during their major depressive episodes. None of the clinical features of major depression differed significantly in frequency among depressed Alzheimer's disease patients with mild, moderate, or severe dementia. Concurrent psychotic symptoms progressively increased with dementia severity. CONCLUSIONS: The high rate of major depressive episodes that occur after the onset of cognitive impairment among patients with Alzheimer's disease (the majority of whom had no premorbid history of major depression), common emergence in the early stages of dementia when symptoms of cognitive impairment are least likely to contribute to the syndromal diagnosis of major depression, and differences in the clinical presentations of the major depressive episodes of Alzheimer's disease patients and nondemented elderly comparison subjects, all support the validity of the major depressive syndrome of Alzheimer's disease. Our findings suggest that the major depressive syndrome of Alzheimer's disease may be among the most common mood disorders of older adults. PMID- 12727689 TI - Amyloid beta pathology in Alzheimer's disease and schizophrenia. AB - OBJECTIVE: Severe cognitive impairment is common in elderly patients with schizophrenia. Alzheimer's disease is the main cause of dementia among the elderly. Biochemical and genetic studies suggest that amyloid beta-peptide is central in Alzheimer's disease. The authors examined the possible involvement of amyloid beta-peptide in cognitive impairment in schizophrenia. METHOD: Specific antibodies against two major forms of amyloid beta-peptide, Abetax-40 and Abetax 42, were used in sandwich enzyme-linked immunosorbent assays to determine the levels of amyloid beta-peptide in postmortem brain samples from Alzheimer's disease patients (N=10), normal elderly comparison subjects (N=11), and schizophrenia patients with (N=7) or without (N=26) Alzheimer's disease. RESULTS: The levels of amyloid beta-peptide were highest in the Alzheimer's disease patients, followed by the patients with schizophrenia and comparison subjects. The mean Abetax-42 level in the schizophrenia patients without Alzheimer's disease was similar to that in the comparison subjects, but the level in the schizophrenia patients with Alzheimer's disease was significantly higher than in those without Alzheimer's disease or the comparison subjects. The Abetax-42 level in the schizophrenia patients with Alzheimer's disease was significantly lower than the level in the Alzheimer's disease cohort. CONCLUSIONS: In contrast to elderly schizophrenia patients with Alzheimer's disease pathology, those without Alzheimer's disease had amyloid beta-peptide levels that were not significantly different from those of normal subjects; hence amyloid beta-peptide does not account for the cognitive deficits in this group. These results suggest that the causes of cognitive impairment in "pure" schizophrenia are different from those in Alzheimer's disease. PMID- 12727690 TI - Lower concentration of hippocampal N-acetylaspartate in familial bipolar I disorder. AB - OBJECTIVE: Previous studies attempting to identify neuropathological alterations in the hippocampus in bipolar disorder have been inconclusive. The objective of this study was to determine if the concentration of N-acetylaspartate, a neuronal and axonal marker, was lower in subjects with familial bipolar I disorder than in healthy comparison subjects, suggesting possible neuronal loss, neuronal dysfunction, or neuropil reduction in bipolar I disorder. METHOD: N acetylaspartate, choline, and creatine in the right and left hippocampus were measured in 15 euthymic male patients with familial bipolar I disorder and 20 healthy male comparison subjects by using proton magnetic resonance spectroscopy ((1)H-MRS). RESULTS: Relative to the comparison group, the patients with bipolar I disorder demonstrated significantly lower concentrations of N-acetylaspartate and creatine but normal choline concentration in both the right and left hippocampus. There were no group or lateralized differences in the percentages of different tissue types within the MRS voxels, suggesting that the hippocampal N acetylaspartate and creatine alterations were not an artifact of variations in tissue types represented in the voxels. There was also a significant negative correlation between N-acetylaspartate concentration in the right hippocampus and illness duration, after adjustment for the effects of age. CONCLUSIONS: This preliminary study provides support for the existence of neuronal loss, neuronal metabolic dysfunction, or interneuronal neuropil reduction in the hippocampal region in male patients with familial bipolar I disorder. The finding of normal hippocampal choline levels in these patients does not provide support for ongoing myelin breakdown or glial cell proliferation in this brain region in familial bipolar I disorder. The significant association between illness duration and N acetylaspartate concentration in the right hippocampus supports the idea that neuronal pathology may increase with disease progression and that this effect may be lateralized, involving the right but not the left hippocampus. PMID- 12727691 TI - Gender differences in prevalence, risk, and clinical correlates of alcoholism comorbidity in bipolar disorder. AB - OBJECTIVE: The prevalence of lifetime alcohol abuse and/or dependence (alcoholism) in patients with bipolar disorder has been reported to be higher than in all other axis I psychiatric diagnoses. This study examined gender specific relationships between alcoholism and bipolar illness, which have previously received little systematic study. METHOD: The prevalence of lifetime alcoholism in 267 outpatients enrolled in the Stanley Foundation Bipolar Network was evaluated by using the Structured Clinical Interview for DSM-IV. Alcoholism and its relationship to retrospectively assessed measures of the course of bipolar illness were evaluated by patient-rated and clinician-administered questionnaires. RESULTS: As in the general population, more men (49%, 57 of 116) than women with bipolar disorder (29%, 44 of 151) met the criteria for lifetime alcoholism. However, the risk of having alcoholism was greater for women with bipolar disorder (odds ratio=7.35) than for men with bipolar disorder (odds ratio=2.77), compared with the general population. Alcoholism was associated with a history of polysubstance use in women with bipolar disorder and with a family history of alcoholism in men with bipolar disorder. CONCLUSIONS: This study suggests that there are gender differences in the prevalence, risk, and clinical correlates of alcoholism in bipolar illness. Although this study is limited by the retrospective assessment of illness variables, the magnitude of these gender specific differences is substantial and warrants further prospective study. PMID- 12727692 TI - The role of psychiatric disorders in predicting drug dependence treatment outcomes. AB - OBJECTIVE: Previous research has demonstrated that psychiatric disorders are common among people who abuse alcohol and drugs, but few studies have examined the relationship of psychiatric disorders to drug treatment outcome. The authors conducted such an examination. METHOD: They successfully reinterviewed 401 drug dependent subjects (94% of the baseline in-treatment sample) and determined their drug abuse status at follow-up 12 months later. RESULTS: Analyses indicated that several baseline psychiatric disorders predicted worse outcomes at follow-up. Major depression predicted using a larger number of substances and having more drug dependence diagnoses and symptoms. Alcohol dependence predicted more dependence diagnoses, antisocial personality disorder predicted using a larger number of substances, and generalized anxiety disorder predicted having more dependence diagnoses. Outcomes among men were more closely associated with psychiatric status than outcomes among women, except for phobias, which predicted a better outcome among women. CONCLUSIONS: These results are unique in their assessment of individuals dependent on illicit substances. Overall, the authors found that women with phobias had better outcomes and that men with psychiatric disorders in general, men with major depression, and men with antisocial personality disorder had worse outcomes. PMID- 12727693 TI - Decreased striatal dopamine D1 receptor-stimulated adenylyl cyclase activity in human methamphetamine users. AB - OBJECTIVE: It has been assumed that some behavioral changes associated with repeated exposure to dopaminergic psychostimulant drugs might be explained by changes in activity of dopamine receptors, including the dopamine D(1) receptor, which is linked by a stimulatory G protein to the effector enzyme adenylyl cyclase. To establish whether dopamine D(1) receptor function might be altered in human methamphetamine users, the authors measured dopamine-stimulated adenylyl cyclase activity in the brain of chronic human users of the drug. METHOD: Adenylyl cyclase activity stimulated by dopamine and by guanylyl-imidodiphosphate (to assess G protein and adenylyl cyclase coupling) was determined in the postmortem brain tissue of 16 methamphetamine users who had used the drug both recently and chronically (i.e., at least 1 year) as well as 21 matched comparison subjects. RESULTS: A 25%-30% decrease in the maximal extent of dopamine stimulation of adenylyl cyclase activity was seen in the striatum (nucleus accumbens, caudate, and putamen) of the methamphetamine users. No changes were found in basal or guanylyl-imidodiphosphate-stimulated enzyme activity. CONCLUSIONS: These data suggest that dopamine receptor function linked to adenylyl cyclase is partially desensitized in the striatum of human methamphetamine users. Decreased dopamine D(1) receptor function might underlie part of the known (drug withdrawal syndrome) or expected (drug tolerance) consequences of methamphetamine exposure in humans. PMID- 12727694 TI - Personality traits and striatal dopamine synthesis capacity in healthy subjects. AB - OBJECTIVE: Neuroimaging and genetic studies suggest that individual differences in the brain dopaminergic system contribute to the normal variability of human personality (e.g., social detachment and novelty seeking). The authors studied whether presynaptic dopamine function is also associated with personality traits. METHOD: Presynaptic dopamine synthesis capacity in the brain was measured with positron emission tomography and [(18)F]fluorodopa in 33 healthy adults, and personality traits were assessed with the Karolinska Scales of Personality. Associations were studied by using a linear regression model controlling for the effects of age and gender on both variables. RESULTS: High scores on two of the anxiety-related personality scales, somatic anxiety and muscular tension, and on one aggressivity-related scale, irritability, were significantly associated with low [(18)F]fluorodopa uptake in the caudate. No statistically significant associations were observed between [(18)F]fluorodopa uptake and the detachment scale or scales related to novelty-seeking behavior (impulsiveness and monotony avoidance). CONCLUSIONS: The results suggest a role for the dopaminergic system in the regulation of anxiety in healthy subjects. Together with previous studies, they also indicate differential involvement of various components of the dopaminergic system in normal and pathological personality traits. PMID- 12727695 TI - An FMRI study of episodic encoding and recognition of words in patients with schizophrenia in remission. AB - OBJECTIVE: Verbal memory deficits are among the most severe cognitive deficits observed in patients with schizophrenia. This study examined patterns of brain activity during episodic encoding and recognition of words in patients with schizophrenia. METHOD: Functional magnetic resonance imaging (fMRI) was used to study regional brain activation in 10 healthy male comparison subjects and 10 male outpatients with schizophrenia during performance of a modified version of the words subtest of Warrington's Recognition Memory Test. RESULTS: Despite having intact performance in word recognition, the patients with schizophrenia had less activation of the right dorsolateral and anterior prefrontal cortex, right anterior cingulate, and left lateral temporal cortex during word encoding, compared with the healthy comparison subjects. During word recognition, the patients had impairments in activation of the bilateral dorsolateral prefrontal and lateral temporal cortices. CONCLUSIONS: Schizophrenia was associated with attenuated frontotemporal activation during episodic encoding and recognition of words. These results from an fMRI study replicate earlier findings derived from a positron emission tomography study. PMID- 12727696 TI - Functional disconnection between the prefrontal and parietal cortices during working memory processing in schizophrenia: a[15(O)]H2O PET study. AB - OBJECTIVE: The prefrontal-parietal circuits have been reported to play an important role in working memory. The purpose of this study was to use direct neuroimaging to address whether interregional correlation in the prefrontal parietal circuits is impaired in schizophrenia. METHOD: [(15)O]H(2)O positron emission tomography was used to compare regional blood flow changes in 12 schizophrenic and 12 healthy comparison subjects during the n-back sequential picture working memory task. Interregional correlation was assessed by correlating the regional activation of the lateral prefrontal area with that of other activated areas in each subject group. RESULTS: Dorsolateral prefrontal activation was observed around the right superior frontal sulcus in the healthy subjects, and ventrolateral prefrontal activation below the right inferior frontal sulcus was observed in the schizophrenic patients. Activation in the right lateral prefrontal cortex was significantly correlated with activation in the bilateral inferior parietal region in the healthy subjects but was not correlated with any regional activation in the patients. CONCLUSIONS: The findings suggest prefrontal-parietal functional disconnection, particularly prefrontal dissociation and abnormal prefrontal-parietal interaction, during working memory processing in schizophrenia. PMID- 12727697 TI - MRI and PET study of deficits in hippocampal structure and function in women with childhood sexual abuse and posttraumatic stress disorder. AB - OBJECTIVE: Animal studies have suggested that early stress is associated with alterations in the hippocampus, a brain area that plays a critical role in learning and memory. The purpose of this study was to measure both hippocampal structure and function in women with and without early childhood sexual abuse and the diagnosis of posttraumatic stress disorder (PTSD). METHOD: Thirty-three women participated in this study, including women with early childhood sexual abuse and PTSD (N=10), women with abuse without PTSD (N=12), and women without abuse or PTSD (N=11). Hippocampal volume was measured with magnetic resonance imaging in all subjects, and hippocampal function during the performance of hippocampal based verbal declarative memory tasks was measured by using positron emission tomography in abused women with and without PTSD. RESULTS: A failure of hippocampal activation and 16% smaller volume of the hippocampus were seen in women with abuse and PTSD compared to women with abuse without PTSD. Women with abuse and PTSD had a 19% smaller hippocampal volume relative to women without abuse or PTSD. CONCLUSIONS: These results are consistent with deficits in hippocampal function and structure in abuse-related PTSD. PMID- 12727698 TI - The relationship between childhood physical abuse and suicidality among depressed women: results from a community sample. AB - OBJECTIVE: The aim of this study was to examine the distribution and determinants of suicidal ideation and suicide attempts in a community sample of women with major depressive disorder. Variables of interest included childhood physical abuse, individual and familial psychiatric history, and sociodemographic factors. METHOD: Women (age 15 to 64 years) from a community sample who met criteria for major depressive disorder (lifetime prevalence) (N=347) were assessed with a modified version of the Composite International Diagnostic Interview. Childhood physical abuse history was collected through a self-administered questionnaire. RESULTS: Approximately one-quarter (23.9%, N=83) of the women with major depressive disorder reported that they had made a suicide attempt, and more than half (55.6%, N=193) had experienced suicidal ideation. A history of suicide attempt was most strongly related to the number of comorbid psychiatric disorders (odds ratio=2.04, 95% confidence interval [CI]=1.21-3.43). Suicidal ideation was most strongly associated with a history of childhood physical abuse (odds ratio=2.77, 95% CI=1.26-6.12). CONCLUSIONS: Suicidal ideation and suicide attempts were reported frequently by women with a history of major depressive disorder. Correlates for suicide attempts showed some differences from those for ideation. This finding is of clinical importance, since it relates to the identification of individuals at risk for suicidal behavior. PMID- 12727699 TI - Family disruption in childhood and risk of adult depression. AB - OBJECTIVE: The authors examined the risk that family disruption and low socioeconomic status in early childhood confer on the onset of major depression in adulthood. METHOD: Participants were 1,104 offspring of mothers enrolled during pregnancy in the Providence, R.I., site of the National Collaborative Perinatal Project. Measures of childhood family disruption and socioeconomic status were obtained before birth and at age 7. Structured diagnostic interviews were used to assess respondents' lifetime history of major depressive episode between the ages of 18 and 39. Survival analysis was used to identify childhood risks for depression onset. RESULTS: Parental divorce in early childhood was associated with a higher lifetime risk of depression among subjects whose mothers did not remarry as well as among subjects whose mothers remarried. These effects were more pronounced when accompanied by high levels of parental conflict. Independent of the respondents' adult socioeconomic status, low socioeconomic status in childhood predicted an elevated risk of depression. CONCLUSIONS: Family disruption and low socioeconomic status in early childhood increase the long-term risk for major depression. Reducing childhood disadvantages may be one avenue for prevention of depression. Identification of modifiable pathways linking aspects of the early childhood environment to adult mental health is needed to mitigate the long-term consequences of childhood disadvantage. PMID- 12727700 TI - Risk of repeat offending among violent female offenders with psychotic and personality disorders. AB - OBJECTIVE: The aim of this study was to examine the rate of criminal recidivism among female homicide offenders evaluated by forensic psychiatrists, to compare this rate with that of other violent female offenders, and to analyze the explanatory variables of recidivism. METHOD: This was a retrospective study of all women (N=132) sent for forensic psychiatric examination after being convicted of homicide or attempted homicide in Finland during 1982-1992; subjects were followed up until mid-1999. Data were collected from the national crime register, the prisoner record, and Statistics Finland. The authors compared the rate of violent repeat offending in this group with that of other violent women and analyzed the explanatory variables of recidivism. RESULTS: During the follow-up period, 23% of the study group committed a repeat offense, 15% of which were violent and 3% of which were homicides. Almost half of all repeat offenses occurred within the first 2 years after the index offense. There was no statistically significant difference in violent recidivism between the study group and other violent female offenders. Of those who committed repeat offenses, 81% were diagnosed with a personality disorder, and 10% were diagnosed with psychosis. Criminality prior to the index event, alcohol or drug dependency, and young age significantly raised the risk and rapidity of further offenses. CONCLUSIONS: The risk of recidivism was high in this study group yet was similar to that of other violent female offenders. The risk was high very early after release. It seems that women and men who are violent and have personality disorders are comparable in their risk of recidivism. PMID- 12727701 TI - Personality diagnoses in adolescence: DSM-IV axis II diagnoses and an empirically derived alternative. AB - OBJECTIVE: The study of personality pathology in adolescence is in its infancy. This article examined the applicability and limits of DSM-IV axis II personality disorder diagnoses in adolescents, assessed the validity of a method for assessing adolescent personality pathology, and began to develop an empirically grounded classification. METHOD: A total of 296 randomly selected clinicians described a patient age 14-18 in treatment for maladaptive personality patterns using axis II ratings scales and the Shedler-Westen Assessment Procedure-200 for Adolescents (SWAP-200-A), a Q-sort instrument for assessing adolescent personality pathology. After examining the nature and frequency of axis II disorders in the sample, the authors used Q-factor analysis to identify naturally occurring groupings of patients on the basis of shared personality features. RESULTS: Axis II diagnoses in adolescents resembled those in adults, although application of DSM-IV criteria appeared to overdiagnose antisocial and avoidant personality disorder in adolescents. Q analysis with the SWAP-200-A isolated five personality disorders (antisocial-psychopathic, emotionally dysregulated, avoidant-constricted, narcissistic, and histrionic) and one personality style. Patients' dimensional scores on each diagnostic prototype showed predictable associations with ratings of current axis II disorders, measures of adaptive functioning, and symptoms assessed with the Child Behavior Checklist. CONCLUSIONS: With some exceptions, personality pathology in adolescence resembles that in adults and is diagnosable in adolescents ages 14-18. Categories and criteria developed for adults may not be the optimal way of diagnosing adolescents. Data from samples of adolescents may prove useful in developing an empirically and clinically grounded classification of personality pathology in adolescents. PMID- 12727702 TI - The intermediate-term outcome of Chinese patients with anorexia nervosa in Hong Kong. AB - OBJECTIVE: The authors determined the intermediate-term outcome of anorexia nervosa for Chinese patients in Hong Kong. METHOD: A consecutive series of 88 patients who fulfilled DSM-III-R criteria for typical (i.e., fat phobic [N=63]) and atypical (i.e., no fat phobia [N=25]) anorexia nervosa were contacted at least 4 years after onset of their illness for semistructured and self-rated assessments of outcome. RESULTS: Three patients (3.4%) died; the mortality ratio for this group against the expected standard for subjects of similar age and gender was 10.5 to 1. Eighty (94.1%) of the remaining 85 patients were successfully traced 9.0 years after onset of their illness. Good, intermediate, and poor outcomes were seen in 61.8%, 32.9%, and 5.3% of the subjects, respectively. Anorexia nervosa, bulimia nervosa, or eating disorder not otherwise specified was exhibited by 55.0% of the subjects (N=44). Lifetime depressive (81.6%) and anxiety (27.6%) disorders were common. Older age at onset and the presence of fat phobia independently predicted poor outcome. Patients with atypical anorexia nervosa were symptomatically stable, less likely to demonstrate bulimia, and had a better eating disorder outcome than patients with typical anorexia nervosa. CONCLUSIONS: The outcome profile of Chinese patients supported the cross-cultural disease validity of anorexia nervosa. The cultural fear of fatness not only shaped the manifest content but also added to the chronicity of the illness. PMID- 12727703 TI - Self-help for bulimia nervosa: a randomized controlled trial. AB - OBJECTIVE: The authors examined the effectiveness of unguided self-help as a first step in the treatment of bulimia nervosa. METHOD: A total of 85 women with bulimia nervosa who were on a waiting list for treatment at a hospital-based clinic participated. The patients were randomly assigned to receive one of two self-help manuals or to a waiting list control condition for 8 weeks. One of the self-help manuals addressed the specific symptoms of bulimia nervosa (cognitive behavior self-help), while the other focused on self-assertion skills (nonspecific self-help). RESULTS: Twenty patients (23.5%) dropped out of the study. The data were analyzed with intention-to-treat analysis. Although the group-by-time interaction for binge eating and purging was not statistically significant, simple effects showed that there was a significant reduction in symptom frequency in both self-help conditions at posttreatment but not in the waiting list condition. There were no statistically significant changes in levels of dietary restraint, eating concerns, concerns about shape and weight, or general psychopathology. A greater proportion of patients in the cognitive behavior self-help (53.6%) and nonspecific self-help (50.0%) conditions reported at least a 50% reduction in binge eating or purging at posttreatment, compared with the waiting list condition (31.0%). A lower baseline knowledge about eating disorders, more problems with intimacy, and higher compulsivity scores predicted a better response. CONCLUSIONS: The findings suggest that a subgroup of patients with bulimia nervosa may benefit from unguided self-help as a first step in their treatment. Cognitive behavior self-help and nonspecific self-help had equivalent effects. PMID- 12727704 TI - Youth hate crimes: identification, prevention, and intervention. AB - OBJECTIVE: Youth hate crimes are a societal problem in which young people turn extremist ideas into acts of violence. To develop methods for prevention, early identification, and intervention, mental health professionals must have an awareness and understanding of this issue. To provide a basis for developing such an understanding, the authors review the current research related to youth hate crimes. METHOD: The authors review the literature primarily from the past 10 years on youth hate crimes. RESULTS: Studies have established that most hate crimes are committed by single or small groups of young males unaffiliated with organized hate groups. Although limited information is available about the causative factors of hatred, a variety of prevention and intervention strategies have been employed. Yet, little has been done to evaluate these various initiatives. Unfortunately, there is a paucity of literature available to guide mental health professionals in the identification, evaluation, and treatment of offenders, despite increasing concerns and awareness regarding the profound consequences of acts of hatred and extremism. CONCLUSIONS: Heightened public awareness and greater understanding of the epidemiology and nature of hate crimes is necessary if perpetrators are to be recognized and effective interventions developed. To achieve this goal, databases of juvenile hate crimes must be developed nationwide, and the success of preventive, educational, and alternative sentencing programs must be assessed. Mental health professionals play a critical role in the detection and treatment of juvenile perpetrators, and it is incumbent upon them to develop interventions for individuals and communities affected by hate crimes. PMID- 12727705 TI - Toward a neuropsychological theory of antidepressant drug action: increase in positive emotional bias after potentiation of norepinephrine activity. AB - OBJECTIVE: Antidepressants that increase serotonin or norepinephrine in the brain are effective in treating depression, but there is no neuropsychological account of how these changes relieve depressive states. Cognitive theories suggest that biases in information processing lead depressed patients to make unrealistically negative judgments about themselves and the world. METHOD: A single dose of the noradrenergic antidepressant reboxetine or placebo was administered to 24 healthy volunteers. Effects on emotional processing were assessed through facial expression recognition, emotional categorization, and emotional memory. RESULTS: On the three measures, reboxetine biased perception toward positive, rather than negative, information in the absence of changes in nonemotional performance or mood. CONCLUSIONS: These results suggest that a single dose of an antidepressant can increase the processing of positively valenced material in nondepressed volunteers. Antidepressants may therefore work in a manner similar to that of psychological treatments that aim to redress negative biases in information processing. PMID- 12727706 TI - Placental passage of antidepressant medications. AB - OBJECTIVE: This study determined the placental transfer of antidepressants and their metabolites. METHOD: A total of 38 pregnant women taking citalopram, fluoxetine, paroxetine, or sertraline participated. Maternal and umbilical cord blood samples were obtained to determine antidepressant and metabolite concentrations. RESULTS: Antidepressant and metabolite concentrations were detectable in 86.8% of umbilical cord samples. The mean ratios of umbilical cord to maternal serum concentrations ranged from 0.29 to 0.89. The lowest ratios were for sertraline and paroxetine; the highest were for citalopram and fluoxetine. Maternal doses of sertraline and fluoxetine correlated with umbilical cord concentrations of these medications. CONCLUSIONS: Umbilical cord concentrations of antidepressants and their metabolites were almost invariably lower than corresponding maternal concentrations. Maternal doses predicted umbilical concentrations of fluoxetine and sertraline. Mean umbilical cord to maternal serum ratios were significantly lower for sertraline than fluoxetine, suggesting that sertraline may produce less fetal medication exposure than fluoxetine near delivery. PMID- 12727707 TI - A double-blind, placebo-controlled study of the omega-3 fatty acid docosahexaenoic acid in the treatment of major depression. AB - OBJECTIVE: This study was an evaluation of the omega-3 fatty acid docosahexaenoic acid (DHA) for the treatment of major depression. METHOD: Thirty-six depressed patients were randomly assigned to receive DHA, 2 g/day, or placebo for 6 weeks. Response was defined a priori as a > or =50% reduction in the score on the Montgomery-Asberg Depression Rating Scale. Thirty-five participants were evaluable; 18 received DHA, and 17 received placebo. RESULTS: Response rates were 27.8% in the DHA group and 23.5% in the placebo group. The difference in response rates between groups did not reach statistical significance. CONCLUSIONS: This trial failed to show a significant effect of DHA monotherapy in subjects with major depression. PMID- 12727708 TI - Age at onset in bipolar I affective disorder: further evidence for three subgroups. AB - OBJECTIVE: Preliminary data suggested that there are three subgroups of bipolar affective disorder based on age at onset. The authors sought to replicate those findings and determine the cut-off age of each subgroup. METHOD: Admixture analysis was used to determine the best-fitting model for the observed ages at onset of 368 consecutively admitted patients. The results obtained were compared with those of the previously described model. The authors also investigated whether affected siblings are more likely to belong to the same theoretical age at-onset subgroup as identified by admixture analysis. RESULTS: The existence of three subgroups defined by age at onset was confirmed. The mean ages estimated in this model were 17.4 years (SD=2.3), 25.1 years (SD=6.2), and 40.4 years (SD=11.3). Affected siblings were more likely to belong to the same theoretical subgroup. CONCLUSIONS: There are three age-at-onset subgroups of bipolar patients, and specific familial vulnerability factors might underlie each subgroup. PMID- 12727709 TI - Obsessive-compulsive disorder in adolescent schizophrenia patients. AB - OBJECTIVE: The co-occurrence of obsessive-compulsive disorder (OCD) in adult patients with schizophrenia has been increasingly recognized. However, the rate of OCD comorbidity in adolescent schizophrenia patients has yet to be systematically evaluated. METHOD: The rate of DSM-IV OCD was evaluated in 50 adolescent inpatients with schizophrenia or schizoaffective disorder. The severity of schizophrenia and OCD symptoms was assessed with the Scale for the Assessment of Positive Symptoms, Scale for the Assessment of Negative Symptoms (SANS), and Yale-Brown Obsessive Compulsive Scale. RESULTS: Thirteen schizophrenia patients (26.0%) also met the DSM-IV criteria for OCD. This subgroup scored significantly higher on the SANS subscale for affective flattening or blunting. The total SANS score positively correlated with the total Yale-Brown Obsessive Compulsive Scale score. CONCLUSIONS: A substantial proportion of adolescent schizophrenia inpatients have concomitant OCD. A prospective study is needed to evaluate the clinical course, response to treatment, and prognosis for this complex disorder. PMID- 12727710 TI - Pathways of low-income minority patients to outpatient psychiatric treatment. AB - OBJECTIVE: Low-income outpatients with scheduled appointments ("scheduled patients") were compared with those who sought treatment without appointments ("walk-ins"). METHOD: The charts of scheduled patients and walk-ins at an outpatient mental health clinic serving a low-income group were reviewed to determine sociodemographic and clinical characteristics and patients' pathways to treatment. RESULTS: Walk-ins (N=241) outnumbered scheduled patients (N=141). The two groups differed significantly in type of presenting complaint and source of referral. A higher proportion of walk-ins sought help with social relationships, while more scheduled patients had complaints involving social performance. Self referrals were more common among the scheduled patients, and family members were more likely to have motivated the walk-ins. CONCLUSIONS: The two groups have more in common than might be expected. Scheduled patients are probably more motivated to seek treatment and therefore more likely to initiate appointments. Walk-ins appear to postpone asking for help until their families urge them to do so. PMID- 12727711 TI - Comorbidity of severe psychiatric disorders and substance use disorders among women in jail. AB - OBJECTIVE: This article presents the prevalence, patterns, and sequences of severe psychiatric disorders and substance use disorders among female jail detainees. METHOD: Subjects were a randomly selected, stratified sample of 1,272 female arrestees awaiting trial at the Cook County Department of Corrections in Chicago. Independent clinical research interviewers administered the National Institute of Mental Health Diagnostic Interview Schedule Version III-R to assess comorbid psychiatric disorders and substance use disorders. RESULTS: Eight percent of the women had both a current severe mental disorder and a current substance use disorder. Nearly three-quarters of those with severe mental disorders also met criteria for one or more substance use disorders. CONCLUSIONS: Because most detainees return to their communities in a few days, these findings have implications for treatment of high-risk women throughout the mental health system. PMID- 12727712 TI - Obstetric complications and schizophrenia. PMID- 12727713 TI - Keratoconus and psychosis. PMID- 12727714 TI - Psychotherapy shattered? PMID- 12727716 TI - Primary care and suicide prevention. PMID- 12727718 TI - Neurology, psychiatry, and neuroscience. PMID- 12727719 TI - Neurology, psychiatry, and neuroscience. PMID- 12727720 TI - Neuropsychiatry. PMID- 12727721 TI - Neuropsychiatry. PMID- 12727736 TI - Smoke free hospitals. PMID- 12727738 TI - People missing as a result of armed conflict. PMID- 12727737 TI - Comparing cannabis with tobacco. PMID- 12727739 TI - Biventricular pacing for heart failure. PMID- 12727740 TI - Scientific literature's open sesame? PMID- 12727741 TI - SARS may have peaked in Canada, Hong Kong, and Vietnam. PMID- 12727742 TI - Canada insists that it is a safe place to visit. PMID- 12727743 TI - SARS could still affect the United Kingdom, health secretary warns. PMID- 12727744 TI - Royal Society warns of risks from depleted uranium. PMID- 12727745 TI - Screening for depression in primary care. Chosen tool makes little sense. PMID- 12727747 TI - Monitoring marketing of infant formula feeds. Manufacturers encourage transparent and official monitoring of WHO code. PMID- 12727749 TI - Cardiovascular mortality after radiotherapy for breast cancer. Article showed nothing new. PMID- 12727748 TI - Screening for depression in primary care. Study analysis and conclusions are flawed. PMID- 12727754 TI - Roll Back Malaria campaign still has a long way to go. PMID- 12727758 TI - US food safety regulations are out of date, report says. PMID- 12727760 TI - Vet dies from pneumonia in avian flu case. PMID- 12727762 TI - Trials of AIDS vaccine to start in Switzerland and United Kingdom. PMID- 12727763 TI - Trials of AIDS vaccine to start in India. PMID- 12727766 TI - Long chain polyunsaturated fatty acid supplementation in infant formula and blood pressure in later childhood: follow up of a randomised controlled trial. AB - OBJECTIVE: To determine whether supplementation of infant formula milk with long chain polyunsaturated fatty acids (LCPUFAs) influences blood pressure in later childhood. DESIGN: Follow up of a multicentre, randomised controlled trial. SETTING: Four study centres in Europe. PARTICIPANTS: 147 formula fed children, with a reference group of 88 breastfed children. INTERVENTION: In the original trial newborn infants were randomised to be fed with a formula supplemented with LCPUFAs (n=111) or a formula without LCPUFAs but otherwise nutritionally similar (n=126). In the present follow up study the blood pressure of the children at age 6 years was measured. MAIN OUTCOME MEASURES: Systolic, diastolic, and mean blood pressure. RESULTS: 71 children in the LCPUFA supplementation group (64% of the original group) and 76 children in the non-supplementation group (60%) were enrolled into the follow up study. The LCPUFA group had significantly lower mean blood pressure (mean difference -3.0 mm Hg (95% confidence interval -5.4 mm Hg to -0.5 mm Hg)) and diastolic blood pressure (mean difference -3.6 mm Hg (-6.5 mm Hg to -0.6 mm Hg)) than the non-supplementation group. The diastolic pressure of the breastfed children (n=88 (63%)) was significantly lower than that of the non supplemented formula group but did not differ from the LCPUFA formula group. CONCLUSIONS: Dietary supplementation with LCPUFAs during infancy is associated with lower blood pressure in later childhood. Blood pressure tends to track from childhood into adult life, so early exposure to dietary LCPUFAs may reduce cardiovascular risk in adulthood. PMID- 12727767 TI - Cost effectiveness of ward based non-invasive ventilation for acute exacerbations of chronic obstructive pulmonary disease: economic analysis of randomised controlled trial. AB - OBJECTIVE: To evaluate the cost effectiveness of standard treatment with and without the addition of ward based non-invasive ventilation in patients admitted to hospital with an acute exacerbation of chronic obstructive pulmonary disease. DESIGN: Incremental cost effectiveness analysis of a randomised controlled trial. SETTING: Medical wards in 14 hospitals in the United Kingdom. PARTICIPANTS: The trial comprised 236 patients admitted to hospital with an acute exacerbation of chronic obstructive pulmonary disease and mild to moderate acidosis (pH 7.25 7.35) secondary to respiratory failure. The economic analysis compared the costs of treatment that these patients received after randomisation. MAIN OUTCOME MEASURE: Incremental cost per in-hospital death. RESULTS: 24/118 died in the group receiving standard treatment and 12/118 in the group receiving non-invasive ventilation (P=0.05). Allocation to the group receiving non-invasive ventilation was associated with a reduction in costs of 49362 pounds sterling (78741 dollars; 73109 euros), mainly through reduced use of intensive care units. The incremental cost effectiveness ratio was -645 pounds sterling per death avoided (95% confidence interval -2310 pounds sterling to 386 pounds sterling), indicating a dominant (more effective and less costly) strategy. Modelling of these data indicates that a typical UK hospital providing a non-invasive ventilation service will avoid six deaths and three to nine admissions to intensive care units per year, with an associated cost reduction of 12000-53000 pounds sterling per year. CONCLUSIONS: Non-invasive ventilation is a highly cost effective treatment that both reduced total costs and improved mortality in hospital. PMID- 12727768 TI - Loss of tolerance and overdose mortality after inpatient opiate detoxification: follow up study. PMID- 12727769 TI - Changing prescription patterns for lithium and valproic acid in old age: shifting practice without evidence. PMID- 12727771 TI - Telephone consultations. PMID- 12727770 TI - Understanding influences on smoking in Bangladeshi and Pakistani adults: community based, qualitative study. AB - OBJECTIVE: To gain detailed understanding of influences on smoking behaviour in Bangladeshi and Pakistani communities in the United Kingdom to inform the development of effective and culturally acceptable smoking cessation interventions. DESIGN: Qualitative study using community participatory methods, purposeful sampling, one to one interviews, focus groups, and a grounded approach to data generation and analysis. SETTING: Newcastle upon Tyne, during 2000-2. PARTICIPANTS: 87 men and 54 women aged 18-80 years, smokers and non-smokers, from the Bangladeshi and Pakistani communities. RESULTS: Four dominant, highly inter related themes had an important influence on smoking attitudes and behaviour: gender, age, religion, and tradition. Smoking was a widely accepted practice in Pakistani, and particularly Bangladeshi, men and was associated with socialising, sharing, and male identity. Among women, smoking was associated with stigma and shame. Smoking in women is often hidden from family members. Peer pressure was an important influence on smoking behaviour in younger people, who tended to hide their smoking from elders. There were varied and conflicting interpretations of how acceptable smoking is within the Muslim religion. Tradition, culture, and the family played an important role in nurturing and cultivating norms and values around smoking. CONCLUSION: Although there are some culturally specific contexts for smoking behaviour in Bangladeshi and Pakistani adults-notably the influence of gender and religion-there are also strong similarities with white people, particularly among younger adults. Themes identified should help to inform the development and appropriate targeting of smoking cessation interventions. PMID- 12727772 TI - Cerebral palsy: what parents and doctors want to know. PMID- 12727774 TI - Benign sleep myoclonus in infancy mistaken for epilepsy. PMID- 12727775 TI - The diabetic foot. PMID- 12727776 TI - Ethics of clinical trials from a bayesian and decision analytic perspective: whose equipoise is it anyway? PMID- 12727777 TI - Screening for depression in primary care. Scientific and statistical errors should have been picked up in peer review. PMID- 12727778 TI - Cardiovascular mortality after radiotherapy for breast cancer. Study showed only estimates of use of adjuvant radiotherapy. PMID- 12727779 TI - Monitoring marketing of infant formula feeds. WHO's global strategy is tool to protect breast feeding and child health. PMID- 12727780 TI - Australian Medical Association accepts generic prescribing. PMID- 12727782 TI - Personality and performance during a medical degree. Selecting for extreme personality types is perilous. PMID- 12727783 TI - Autism spectrum disorder is not as certain as implied. PMID- 12727784 TI - Perhaps medicine should now be considered a trade. PMID- 12727785 TI - Clinical environmental medicine. PMID- 12727786 TI - Working as a medical homoeopath. PMID- 12727787 TI - Training in acupuncture. PMID- 12727788 TI - Hypnotherapy: hype or healing? PMID- 12727789 TI - Variability and regulation of O6-alkylguanine-DNA alkyltransferase. AB - O(6)-Alkylguanine-DNA alkyltransferase (ATase) confers resistance to many of the biological effects of certain classes of alkylating agents by repairing the DNA lesions responsible. The role of ATase in the mutagenic and toxic effects of the carcinogenic and antitumour alkylating agents are of interest in relation to the prevention and treatment of cancer in man. In this commentary we specifically focus on the variation in ATase levels and our current understanding of the factors involved in the regulation of ATase expression. PMID- 12727790 TI - Nitric oxide induces cyclooxygenase expression and inhibits cell growth in colon cancer cell lines. AB - The role of nitric oxide (NO) in colon cancer remains controversial. Inducible nitric oxide synthase (iNOS) has been reported to be up regulated and down regulated in colorectal cancer in both animal models and patient tissue samples. Cyclooxygenase-2 (COX-2) is important in colorectal carcinogenesis but its relationship with NO has never been studied in colon cancer. Three colon cancer cell lines (HCA7, HT29 and HCT116) with different COX-2 expression and activities were used to study the effect of the NO donor, S-nitrosoglutathione (GSNO). The effects of GSNO (10-500 micro M) on cell growth, PGE(2) production, COX-1/COX-2 protein expression and cell-cycle distribution were evaluated. GSNO increased PGE(2) production and induced COX-1 and COX-2 protein expression in a dose- and time-dependent manner. Higher concentrations of GSNO also inhibited cell growth and induced apoptosis in all three cell lines, regardless of their COX-2 expression/activities. Inhibition of PGE(2) production did not further improve the inhibitory effect of GSNO. PMID- 12727791 TI - Cells heterozygous for the ApcMin mutation have decreased gap junctional intercellular communication and connexin43 level, and reduced microtubule polymerization. AB - Mutations in the tumour suppressor gene adenomatous polyposis coli (Apc) are early and critical events in the development of colon cancer. In the absence of functional Apc, beta-catenin is not degraded in the cytoplasm and can be transported to the nucleus and turn on transcription of several genes, including the gap junction protein connexin43. Apc also stabilizes microtubules and regulates microtubule polymerization. Changes in Wnt signalling and microtubule function are reported to affect the connexin level. To study the effect of heterozygous Apc mutation we examined gap junctional intercellular communication (GJIC) in IMCE (Immorto-Min colonic epithelium) cells with one mutated Apc allele and in YAMC (Young adult mouse colon) cells with normal Apc function. IMCE cells had only half the GJIC level compared with YAMC cells. RT-PCR showed that both YAMC and IMCE cells express a common complement of seven connexin genes (Cx26, Cx31, Cx39, Cx40, Cx43, Cx45 and Cx50), with an additional Cx29 gene expression in YAMC cells. We found that the Cx43 level was correspondingly lower in IMCE cells as detected by western blotting and immunofluorescence. There were no differences in the level or localization of beta-catenin and the downstream gene E-cadherin between the cells, indicating no activation of the Wnt-signalling pathway in cells with one mutated Apc allele. We also examined the microtubule polymerization rate, and IMCE cells had markedly slower microtubule polymerization than YAMC cells. Hence, it appears that mutation in one Apc allele is sufficient to affect microtubule function, while inactivation of both wild type Apc alleles may be necessary for activation of Wnt signalling. Reduction in GJIC and Cx43 level in IMCE cells may be caused by reduced Cx43 transport as a result of alterations in microtubule function. PMID- 12727792 TI - Proteomic analysis of a neoplastic mouse lung epithelial cell line whose tumorigenicity has been abrogated by transfection with the gap junction structural gene for connexin 43, Gja1. AB - In order to examine how tumorigenicity is abrogated by gap junctional intercellular communication (GJIC), protein expression was analyzed in four related mouse lung epithelial cell lines that vary in their GJIC status and neoplastic potential. Since alterations in protein expression underlie neoplastic behavior, this proteomic analysis provides insights into the molecular pathogenesis of lung cancer. E10, an immortalized but non-tumorigenic cell line derived from alveolar type II pneumocytes, has functional GJIC. E9, a spontaneous transformant of E10, is GJIC-deficient and is tumorigenic upon injection into a syngeneic mouse. Stable transfection of E9 with Gja1, the gene for the gap junctional protein, connexin 43, re-established GJIC and rendered this line (designated E9-2) non-tumorigenic; the vector transfection control line, E9-41, remains tumorigenic. Proteins extracted from these cell lines were separated by two-dimensional electrophoresis (2DE) and visualized by Coomassie blue staining. We consistently observed differential expression of 27 proteins between E10 and E9 and identified 11 of these by peptide mass mapping. The functions of these proteins include stress response, cytoskeletal structure, signal transduction, apoptosis, immune response, pre-mRNA processing, and carbohydrate metabolism. Gja1 transfection affected the concentrations of four of these proteins, viz. PDI, alpha-enolase, aldolase A, and gelsolin-like protein. PDI concentration was most profoundly affected; E10 cells contain twice as much PDI as E9, and PDI was restored to E10-like levels in the E9-2 transfectant line while remaining at E9 like levels in the vector control E9-41 cells. An association between connexin 43 and PDI expression was also observed in a second set of independently derived type II cell lines. The PDI superfamily has multiple cellular roles including chaperoning assembled glycoproteins, regulating the activities of transcription factors, and regulating disulfide bond formation. PMID- 12727793 TI - Characterization and chromosomal instability of novel derived cell lines from a wt-erbB-2 transgenic mouse model. AB - Amplification and overexpression of the erbB-2 (HER-2/neu) proto-oncogene and exposure to the cell cycle mitogenic hormone estrogen (E2) have been associated with mammary tumorigenesis. Phytoestrogens found in soy act as selective estrogen receptor modulators and may also modify mammary carcinogenesis. We have used the wt-erbB-2 transgenic mouse model to study the effects of estrogen and dietary phytoestrogens on erbB-2-associated mammary tumorigenesis. Transgenic mice were treated with short-term E2 or placebo pellets during the early reproductive period and fed a casein or soy diet for life. Mammary tumors from the different treatment groups were used for the derivation of novel cell lines. Comparative genomic hybridization (CGH), flow cytometry, assays of cell proliferation and soft agar cloning were performed to study genomic instability and in vitro characteristics. CGH data were compared with corresponding parental tumors. Mammary tumors exhibited significantly fewer genetic changes than cell lines by CGH. Cell lines from soy-fed animals (that developed tumors with a longer latency) demonstrated the greatest frequency of chromosomal gain and loss. The E2 treated, casein-fed animals (that developed tumors with the shortest latency) had the fewest genetic changes in derived lines by CGH. Nonetheless, E2-associated tumors in vivo and lines in vitro had the most aggressive phenotypes. In addition, over 40% of all derived cell lines, and both tumors from the placebo treated casein-fed mice, exhibited loss of chromosome 4 by CGH. In aggregate, our data suggest that estrogenic signaling influences mammary tumor development in this transgenic mouse model bearing the rat wt-erbB-2 gene. Once induced, tumors and derived lines demonstrate persistent phenotypic characteristics, including tumor aggression and shortened latency in E2-treated mice. Loss of chromosome 4 was commonly identified in derived lines and may have facilitated immortalization or passage in culture. PMID- 12727794 TI - Expression of cyclooxygenase-2 parallels expression of interleukin-1beta, interleukin-6 and NF-kappaB in human colorectal cancer. AB - Elevated expression of cyclooxygenase-2 (COX-2), the inducible isoform of prostaglandin H synthase, has been found in several human cancers, including colorectal cancer (CRC). This appears as a rationale for the chemopreventive effects of non-steroidal anti-inflammatory drugs in CRC. However, the reason for COX-2 overexpression is not fully understood. In cell culture experiments, COX-2 can be induced by proinflammatory cytokines, such as interleukin (IL)-1beta and IL-6. A crucial step in this signalling pathway is thought to be activation of transcription factor NF-kappaB. Based on these findings, we hypothesized an association between COX-2 overexpression and expression of IL-1beta, IL-6 and the NF-kappaB subunit p65 in human CRC. To test the hypothesis, we performed immunohistochemistry for the respective antigens on colorectal cancer specimens, obtained by surgical resections from 21 patients with CRC. Immunohistochemical results were confirmed by examination of protein levels in tissue lysates and nuclear extracts using western blotting. Non-neoplastic tissue specimens resected well outside the tumour border served as controls. COX-2 expression was found to be markedly enhanced in the neoplastic epithelium compared with controls. This was paralleled by a significantly higher expression of IL-1beta, IL-6 and p65. Serial sections revealed consistent cellular colocalizations of respective antigens in the neoplastic epithelium. Statistically, a significant correlation between expression of COX-2 and IL-1beta, IL-6 and p65 was found. Comparable results were obtained for stromal cells like macrophages and myofibroblasts. Further examination of nuclear extracts from CRC-specimens by western blotting confirmed a higher content of p65 protein compared with non-neoplastic control tissues. Therefore, our study provides evidence for an association between expression of COX-2 and IL-1beta, IL-6 and p65 in human CRC. The results are consistent with the thesis that proinflammatory cytokines such as IL-1beta and IL 6 may be accountable for the overexpression of COX-2 in CRC. Finally, the study corroborates a role for NF-kappaB in the control of COX-2 gene transcription in CRC. Given an antiapoptotic role for COX-2 in tumour cells, inhibition of NF kappaB may offer an important strategy to interfere with the development and progression of CRC. PMID- 12727795 TI - TCDD-mediated alterations in the AhR-dependent pathway in Seveso, Italy, 20 years after the accident. AB - Approximately 20 years after the Seveso, Italy, accident we conducted a population-based study to evaluate the impact of 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD) exposure on cancer using mechanistically based biomarkers of dioxin response in humans. TCDD toxic effects are mediated by the aryl hydrocarbon receptor (AhR). We studied the AhR-dependent pathway in lymphocytes from 62 subjects randomly sampled from the highest exposed zones and 59 subjects from the surrounding non-contaminated area, frequency matched for age, gender and smoking. To our knowledge, this is the most comprehensive investigation to date designed to evaluate the key genes in the pathway, including AhR, aryl hydrocarbon receptor nuclear translocator, CYP1A1 and CYP1B1 transcripts and CYP1A1 associated 7-ethoxyresorufin O-deethylase (EROD) activity in a population heavily exposed to dioxin. Current lipid-adjusted plasma TCDD concentrations in these subjects ranged from 3.5 to 90 ng/kg (or p.p.t.) and were negatively associated with AhR mRNA in unstimulated peripheral blood mononuclear cells (P = 0.03). When mitogen-induced lymphocytes were cultured with 10 nM TCDD, all AhR-dependent genes were induced 1.2- to 13-fold. In these cells, plasma TCDD was associated with decreased EROD activity. In addition, there was a strong positive correlation between AhR and CYP1A1 expression (P = 0.001) and between AhR and CYP1B1 expression (P = 0.006). CYP1A1 expression was also strongly correlated with EROD activity (P = 0.001). The analysis of the expression of dioxin inducible genes involved in carcinogenesis may help in determining dose-response relationships for human exposure to dioxin in vivo and in assessing the variability of human response, which may indicate the presence of subjects more susceptible to disease as a result of such exposures. PMID- 12727796 TI - Catechol-O-methyltransferase gene polymorphism and post-menopausal breast cancer risk. AB - Estrogen is involved in breast carcinogenesis. Hypotheses have been raised that its effect is modified by enzymes such as catechol-O-methyltransferase (COMT) that deactivate potentially genotoxic estrogen metabolites. We have investigated the association between the functional genetic Val108/158Met polymorphism in COMT and breast cancer risk in a large population-based case-control study performed in the genetically homogeneous Swedish population. We determined COMT genotype in 1534 women with invasive breast cancer and in 1504 control women and calculated odds ratios (OR) and 95% confidence intervals (CI) from logistic regression models. There was no overall association between COMT genotype and breast cancer risk. However, the L allele was associated with an increased risk for lobular breast cancer, with OR 2.0 (95% CI 1.2-3.5) for HL and 1.7 (95% CI 0.9-3.0) for LL. In exploratory subset analyses, we found no statistically significant interaction, but some indication of a positive association between HL and LL genotypes and breast cancer among women with diabetes mellitus and a negative association among nulliparous women. Based on our findings, COMT activity alone does not seem to play a major role in breast carcinogenesis, but may be of importance in certain histotypes or in conjunction with other exposures. PMID- 12727797 TI - Dietary fruit and vegetables protect against somatic mutation in vivo, but low or high intake of carotenoids does not. AB - Epidemiological studies have demonstrated protective effects of vegetables and fruit on risk of cancer, but underlying mechanisms remain unclear. Intervention studies have in some cases contradicted previous epidemiological evidence, e.g. for beta-carotene supplementation and lung cancer, emphasizing the need for mechanistic data. We assessed in vivo mutagenic effects of several dietary items using the HPRT (hypoxanthine-guanine phosphoribosyl transferase) gene assay with T-lymphocytes from 312 individuals (158 lung cancer cases, 154 population controls), who provided information on diet and smoking habits. HPRT mutant frequency (MF) was significantly decreased in relation to intake of vegetables, citrus fruits and berries, respectively, as well as calculated vitamin C intake from diet. There was a significant U-shaped association with dietary carotenoid intake, with lowest MF near population average carotenoid intakes and higher mutation frequencies both at low and high intakes, and a similar borderline significant association was observed for beta-carotene. Our study is consistent with known diet-cancer associations and provides novel human in vivo mechanistic support for a cancer-protective effect of vegetables and fruit by modulation of somatic mutagenesis. Our results also provide support for the increase in lung cancer risk observed particularly in smokers in studies of beta-carotene supplementation. PMID- 12727798 TI - Relative imbalances in estrogen metabolism and conjugation in breast tissue of women with carcinoma: potential biomarkers of susceptibility to cancer. AB - Exposure to estrogens has been associated with an increased risk of developing breast cancer. Breast biopsy tissues from 49 women without breast cancer (controls) and 28 with breast carcinoma (cases) were analyzed by HPLC with electrochemical detection for 31 estrogen metabolites and catechol estrogen quinone-glutathione conjugates. The levels of estrone and estradiol were higher in cases. More 2-catechol estrogen (CE) than 4-CE was observed in controls, but the 4-CE were three times higher than 2-CE in cases. In addition, the 4-CE were nearly four times higher in cases than in controls. Less O-methylation was observed for the CE in cases. The level of catechol estrogen quinone conjugates in cases was three times that in controls, suggesting in the cases a higher probability for the quinones to react with DNA and generate mutations that may initiate cancer. The levels of 4-CE and quinone conjugates were highly significant predictors of breast cancer. These results suggest that some catechol estrogen metabolites and conjugates could serve as biomarkers to predict risk of breast cancer. PMID- 12727799 TI - K-ras oncogene mutations in sporadic colorectal cancer in The Netherlands Cohort Study. AB - Activation of K-ras oncogene has been implicated in colorectal carcinogenesis, being mutated in 30-60% of the adenocarcinomas. In this study, 737 incident colorectal cancer (CRC) patients, originating from 120 852 men and women (55-69 years at baseline) participating in the Netherlands Cohort Study (NLCS), were studied in order to evaluate subgroups with respect to K-ras mutation status. Mutation analysis of the exon 1 fragment of the K-ras oncogene, spanning codons 8 29, was performed on archival colorectal adenocarcinoma samples of all patients using macrodissection, nested PCR and direct sequencing of purified fragments. The method of mutation detection was validated by the confirmation of reported K ras status in CRC cell lines, a good correlation between fresh-frozen and routinely fixed, paraffin-embedded tissue, a detection limit of 5% mutated DNA and a good reproducibility. Various types of K-ras mutations were evaluated with respect to tumour sub-localization, Dukes' stage and tumour differentiation. In 37% (271/737) of the patients, the exon 1 fragment of K-ras gene was found to be mutated. The predominant mutations are G>A transitions and G>T transversions, and codons 12 and 13 are the most frequently affected codons. Patients with a rectal tumour were found to have the highest frequency of G>T transversions as compared with patients with a colon or rectosigmoid tumour. This difference appeared to be confined to women with a rectal tumour harbouring G>T transversions. No significant differences were observed for Dukes' stage with respect to types of K ras mutation, which does not support direct involvement of the K-ras oncogene in adenocarcinoma progression. The equal distribution of K-ras mutations among cases with or without a family history of colorectal cancer argues against an important role for this mutation in familial colorectal cancer, and could imply that K-ras mutations are more probably involved in environmental mechanisms of colorectal carcinogenesis. PMID- 12727800 TI - Susceptibility to prostate cancer: studies on interactions between UVR exposure and skin type. AB - Recent studies have proposed that exposure to ultraviolet radiation (UVR) protects against development of some internal cancers including that in prostate. This effect may be mediated by UVR-induced cutaneous synthesis of vitamin D. It is also proposed that ability to pigment in response to UVR will influence susceptibility to prostate cancer through its effects on vitamin D synthesis. We wished to determine first, whether ability to pigment, as assessed by skin type, influences the extent of exposure to UVR, secondly, whether skin type is associated with prostate cancer susceptibility and thirdly, whether such an effect is mediated by the extent of UVR exposure. We studied 453 prostatic adenocarcinoma and 312 benign prostatic hypertrophy (BPH) patients using a validated questionnaire to assess two parameters of exposure; months of cumulative exposure per year and adult sunbathing score. We used analysis of variance to show that cancer cases with sun-sensitive skin (skin type 1) had lower cumulative exposure per year (P = 0.014) and sunbathing scores (P < 0.0001) than those with type 4, possibly because of a tendency to avoid exposure. Further, cumulative exposure per year and sunbathing score were significantly lower in cancer compared with BPH patients (P < 0.001 and P < 0.001, respectively). While the proportion of subjects with skin types 1 and 2 was lower in cancer than BPH patients, these were not significantly different (logistic regression analysis, skin type 1 versus type 4; P = 0.11). We used recursive partitioning to determine if skin type influenced susceptibility to prostate cancer in subgroups stratified by exposure. Analysis of the data showed that in men with low sunbathing scores, skin type 1 conferred protection compared with skin types 2-4 (OR = 4.78, 95% CI 3.01-8.25, P < 0.0009). These findings indicate that susceptibility to prostate cancer is in part determined by extent of exposure to UVR and that ability to pigment mediates this effect. PMID- 12727802 TI - Characterization of the rat aflatoxin B1 aldehyde reductase gene, AKR7A1. Structure and chromosomal localization of AKR7A1 as well as identification of antioxidant response elements in the gene promoter. AB - Rat aflatoxin B(1) aldehyde reductase (called AFAR1 or AKR7A1) is a member of the aldo-keto reductase 7 family, which metabolizes the environmental carcinogen aflatoxin B(1). The expression of this enzyme is markedly increased in rat liver by cancer chemopreventive agents, many of which are believed to regulate gene expression through the antioxidant response element (ARE). In order to understand how this gene is regulated, two overlapping genomic clones have been isolated that contain most of the coding region for the enzyme; together they encompass 14.1 kb of DNA. Characterization of these clones has shown that rat AFAR1 is approximately 8 kb long and comprises seven exons and six introns. The seven exons are between 97 and 380 bp in size. The introns range in size from 194 bp to approximately 2.9 kb. Fluorescent in situ hybridization localized AFAR1 to rat chromosome 5q36.5, a region that is syntenic with human chromosome 1p35-1p36.1 where AKR7A2 resides. The transcriptional start site (TSS) was determined, using 5'-rapid amplification of cDNA ends, to be an A nucleotide 73 bp upstream from the ATG initiation codon. The 5'-flanking region of AFAR1 was isolated by polymerase chain reaction-based genome walking, and resulted in the isolation of approximately 900 bp of genomic DNA upstream from the TSS. Use of a gene expression reporter assay demonstrated that this cloned 5'-flanking region of AFAR1 could support transcription in the rat liver 34 (RL34) epithelial cell line. Within this upstream region of the promoter, a substantial number of sequences were found that are closely similar, but not identical, to the 'core' ARE consensus sequence. Between nucleotides -810 and -106 bp from the TSS 16 ARE related sequences were identified. Four of these putative enhancers lay between 389 and -355 bp, and the motif 5'-GAGTGAG-3' was repeated three times within the 35 bp region. PMID- 12727801 TI - Evaluation of 4-aminobiphenyl-DNA adducts in human breast cancer: the influence of tobacco smoke. AB - Breast cancer is one of the major cancers around the world but its etiology is still not well understood. Only approximately 50% of the disease is associated with known risk factors including highly penetrant genes and lifestyle factors. Thus, environmental carcinogens may play an important role in the etiology of breast cancer. The arylamine 4-aminobiphenyl (4-ABP) is a tobacco smoke constituent, an environmental contaminant, and a well-established bladder carcinogen in rodents and humans. In this study, we investigated the role of 4 ABP in the etiology of human breast cancer by measuring 4-ABP-DNA adducts using a monoclonal antibody based immunoperoxidase method that had been validated by comparison with gas chromatography/mass spectroscopy analysis of liver tissues from 4-ABP-treated mice. Adducts were analyzed in 150 paraffin-embedded breast tumors and in 55 adjacent normal tissues collected from cases in the Long Island Breast Cancer Study Project. The role of polymorphisms in genes involved in the metabolism of 4-ABP including N-acetyl transferase 2 (NAT2), cytochrome P4501A2 (CYP1A2) and glutathione S-transferase M1 (GSTM1) and the nucleotide excision repair gene XPD was also explored in the same patients. The mean log-transformed relative staining intensity for 4-ABP-DNA adducts was higher in normal (5.93 +/- 0.54) than in the corresponding tumor (5.44 +/- 0.62, P < 0.0001) tissues. However, a highly significant positive correlation was observed between the levels of 4-ABP-DNA in both tissues (r = 0.72, P < 0.0001). Smoking status was correlated with the levels of 4-ABP-DNA in tumor adjacent normal tissues with a significant linear trend (P = 0.04) for current, former and never smokers; adducts were not related to smoking status in tumor tissues. No correlation was observed between the levels of 4-ABP-DNA and polymorphisms in the genes analyzed even when subjects were stratified by smoking status. These results demonstrate that smoking is associated with increased levels of 4-ABP-DNA adducts in human mammary tissue. In this study, genetic polymorphisms did not significantly affect the formation of 4-ABP-DNA adducts in breast cancer cases, perhaps due to the small number of samples. PMID- 12727803 TI - Biomarkers of dietary intake of micronutrients modulate DNA adduct levels in healthy adults. AB - DNA adducts, a reliable indicator of internal dose exposure to genotoxic agents and, possibly, of cancer risk, have been shown to be modulated by diet, particularly by the consumption of fresh fruit and vegetables, and by the intake of antioxidants (Palli et al., 2000, Int. J. Cancer, 87, 444-451). We have therefore investigated the association between DNA adducts in peripheral leukocytes and plasma levels of selected micronutrients, also taking into account the role of metabolic polymorphisms and smoking history, in a large independent random sample of volunteers enrolled in the prospective study EPIC-Italy (approximately 110 subjects from each of the three main geographical study areas, Northern, Central and Southern Italy). DNA adducts and five polymorphic metabolic genotypes were determined in peripheral leukocytes using the (32)P-post-labelling technique and PCR methods. Plasma levels of six carotenoids, retinol and alpha- and gamma-tocopherol were determined in the same blood sample. Among 331 subjects, 78.3% had detectable levels of DNA adducts (mean 7.46 +/- 0.48 per 10(9) nucleotides). Vitamin supplementation was reported by only a few subjects (3.9%). Strong inverse associations emerged between levels of DNA adducts and plasma retinol (P = 0.02), alpha-tocopherol (P = 0.04) and gamma-tocopherol (P = 0.03), but not carotenoids (except a borderline inverse association with beta carotene, P = 0.08). An inverse significant association with plasma levels of retinol and gamma-tocopherol persisted in the subgroup of non-smokers, whereas a negative association with alpha-tocopherol emerged only in smokers. DNA adduct levels did not show any significant variation according to analyzed genotypes. Stratification by GSTM1 genotype, however, showed a significant negative association between DNA adduct levels and plasma levels of alpha- (P = 0.02) and beta-carotene (P = 0.02) in subjects with the GSTM1 null genotype. Our results confirm that biomarkers of dietary intake of antioxidants significantly modulate DNA adducts and suggest specific inverse associations between DNA adduct levels and antioxidant concentrations among GSTM1 null subjects and smokers. PMID- 12727804 TI - Global alteration of gene expression in human keratinocytes by inorganic arsenic. AB - Alteration of gene expression by inorganic arsenic has been studied in cultured human keratinocytes derived from normal epidermis, a premalignant lesion and a malignant tumor. The purpose was to find whether these cells displayed common alterations in gene expression that might elucidate the mechanism of arsenic action. Global analysis of approximately 12 000 genes by microarray showed that approximately 30% were expressed. Of these, transcription of a substantial fraction (up to 12%) was altered, nearly twice as many being suppressed as stimulated by 2-fold or more at 2 micro M sodium arsenite or 6 micro M arsenate, which did not affect cell growth. At 0.67 micro M arsenite (50 p.p.b.), effects on transcription were less pronounced but clearly evident. Genes whose transcription was altered in common among all the treated keratinocytes included those induced by reactive oxygen, of which heme oxygenase-1 displayed the highest fold induction. Genes indicative of reactive oxygen generation were detected at the earliest time examined, raising the possibility this feature drives subsequent cellular responses. Unlike some agents that produced transient induction of heme oxygenase-1, arsenicals produced sustained induction. Comparison with other agents producing reactive oxygen in the cells, as reflected in heme oxygenase-1 induction, suggested cellular differentiation was suppressed by sustained but not transient generation of reactive oxygen. Sustained global changes in gene expression were seen in target cells treated chronically with inorganic arsenic at concentrations consumed by millions of humans in contaminated drinking water. PMID- 12727805 TI - Changes in global gene and protein expression during early mouse liver carcinogenesis induced by non-genotoxic model carcinogens oxazepam and Wyeth 14,643. AB - We hypothesized that the mouse liver tumor response to non-genotoxic carcinogens would involve some common early gene and protein expression changes that could ultimately be used to predict chemical hepatocarcinogenesis. In order to identify a panel of genes to test, we analyzed global differences in gene and protein expression in livers from B6C3F1 mice following dietary treatment with two rodent carcinogens, the benzodiazepine anti-anxiety drug oxazepam (2500 p.p.m.) and the hypolipidemic agent Wyeth (Wy)-14,643 (500 p.p.m.) compared with livers from untreated mice. Male mice were exposed for 2 weeks and 1, 3 or 6 months to oxazepam or Wy-14,643 in an age-matched study design. By histopathological evaluation, no liver preneoplastic foci or tumors were detected at 6 months in treated or control groups. By cDNA microarray analysis [NIEHS Mouse Chip (8700 genes); n = 3 individual livers/group, four hybridizations/sample], expression of 36 genes or 220 genes were changed relative to control livers following 6 months of oxazepam or Wy-14,643 treatment, respectively. To obtain a more comprehensive picture of gene/protein expression changes, we also conducted a proteomics study by 2D-gel electrophoresis followed by matrix assisted laser desorption/ionization mass spectrometry on cytoplasmic, nuclear, and microsomal subcellular fractions of the same liver samples utilized for the cDNA microarray analysis. Real-time PCR, western blot analysis and immunohistochemistry were utilized for validation and to expand the results to other time points. Cyp2b20, growth arrest- and damage-inducible gene beta (Gadd45beta), tumor necrosis factor alpha-induced protein 2 and insulin-like growth factor binding protein 1 (Igfbp5) genes and proteins were upregulated by oxazepam, and Cyp2b20, Cyclin D1, proliferating cell nuclear antigen, Igfbp5, Gadd45beta and cell death-inducing DNA fragmentation factor alpha subunit-like effector A exhibited higher expression after Wy-14,643 treatment. Most of these genes/proteins were also deregulated at 2 weeks. There appeared to be more distinct than common changes in the expression of carcinogenesis-related genes/proteins between the two compounds, suggesting that the major carcinogenic pathways are different for these compounds and may be distinct for different chemical classes. PMID- 12727806 TI - Chromium(VI) exposure enhances polycyclic aromatic hydrocarbon-DNA binding at the p53 gene in human lung cells. AB - Chromium(VI) [Cr(VI)] is a ubiquitous environmental and industrial contaminant. Cr(VI) exposure is strongly associated with a higher incidence of human lung cancer, but the mechanism of Cr(VI) carcinogenicity remains unclear. Cigarette smoking has been known as the prominent cause of lung cancer, and polycyclic aromatic hydrocarbons (PAHs), the major carcinogens in cigarette smoke, have been suggested as being responsible for the initiation and development of lung cancer. It has been reported that lung cancer from workers exposed to Cr(VI) has a high percentage of G to T transversion mutations in the non-transcribed strand of the p53 gene, a hallmark of PAH-induced mutation. Cr(VI) is a weak mutagen although it can induce a high percentage of G to T transversion mutations. These results raise the possibility that Cr(VI) may enhance PAH binding at the p53 gene in lung tissue. To test this possibility, we have determined the effect of Cr(VI) exposure on benzo[a]pyrene diol epoxides (BPDE)-DNA binding at total genomic DNA level and at the p53 gene in normal human lung fibroblast cells. We found that in lung cells Cr(VI) pre-exposure does not affect the BPDE-DNA binding at the total genomic DNA level or at exons 5, 6 and 9 of the p53 gene; however, it greatly enhances BPDE-DNA binding at exons 7 and 8 of the p53 gene, especially at mutational hotspots of lung cancer: codons 248, 273 and 282 of the p53 gene. No enhancement of BPDE-DNA binding in the p53 was observed when naked genomic DNA isolated from Cr(VI)-exposed cells was modified with BPDE in vitro. These results suggest that Cr(VI) exposure may enhance chromatin structure-dependent carcinogen DNA binding. This effect may contribute to the synergism of Cr(VI) and BPDE on mutagenesis and cell transformation, and may also contribute to the higher incidence of lung cancer in Cr(VI)-exposed populations. PMID- 12727807 TI - Hemoglobin and DNA adducts in rats exposed to 2-nitrotoluene. AB - 2-Nitrotoluene (2NT) is an important commercial chemical intermediate. A recent National Toxicology Programme (NTP)-study demonstrated clear evidence of carcinogenic activity of 2NT in rats. In the present study male WELS-Fohm rats were dosed chronically with 2NT, 5 days a week for 12 weeks. Hemoglobin (Hb) adducts and hepatic DNA adducts were analyzed. After mild base treatment of Hb, 2 methylaniline (2MA) was released and quantified using gas chromatography/mass spectrometry. 2'-Deoxyguanosine (dG) and 2'-deoxyadenosine (dA) adducts of 2MA were found in hepatic DNA using electrospray-mass spectrometry (ESI-MS/MS). The dG adduct found in vivo did not co-elute with N-(2'-deoxyguanosine-8-yl)-2 methylaniline which is the expected adduct for arylamines. The dG adduct detected in the dosed rats was not present in calf thymus-DNA (ct-DNA) modified in vitro with N-acetoxy-2MA. The dA adduct detected in rats was a very minor product in ct DNA modified in vitro. The dG and dA adducts found in the 2NT-dosed rats increased with the dose. The same increase was seen for the Hb adduct levels measured in the same animals. The increase of DNA and Hb adduct levels were supralinear. There was a very strong linear relationship between the level of dG 2MA adducts and dA-2MA adducts in hepatic DNA from rats administered 2NT over the whole dose range studied (r(2) = 0.9). A strong linear relationship also existed between the level of dG-2MA or dA-2MA adducts, in hepatic DNA, and Hb adducts, over the whole dose range (r(2) > or = 0.9). Thus, there was strong evidence to support the notion that Hb adducts were an effective surrogate marker for the hepatic DNA damage of rats chronically administered 2NT. PMID- 12727808 TI - Real-time vital optical imaging of precancer using anti-epidermal growth factor receptor antibodies conjugated to gold nanoparticles. AB - Recent developments in photonic technology provide the ability to noninvasively image cells in vivo; these new cellular imaging technologies have the potential to dramatically improve the prevention, detection, and therapy of epithelial cancers. Endoscope-compatible microscopies, such as optical coherence tomography and reflectance confocal microscopy, image reflected light, providing a three dimensional picture of tissue microanatomy with excellent spatial resolution (1 10 micro m). However, their ability to image molecular biomarkers associated with cancer is limited. Here, we describe a new class of molecular specific contrast agents for vital reflectance imaging based on gold nanoparticles attached to probe molecules with high affinity for specific cellular biomarkers. The application of gold bioconjugates for vital imaging of precancers is demonstrated using cancer cell suspensions, three-dimensional cell cultures, and normal and neoplastic fresh cervical biopsies. We show that gold conjugates can be delivered topically for imaging throughout the whole epithelium. These contrast agents have potential to extend the ability of vital reflectance microscopies for in vivo molecular imaging. They can potentially enable combined screening, detection, and therapy of disease using inexpensive imaging systems; such tools could allow mass screening of diseases such as cancer in resource-poor settings. PMID- 12727809 TI - The mutant K-ras oncogene causes pancreatic periductal lymphocytic infiltration and gastric mucous neck cell hyperplasia in transgenic mice. AB - A frequent genetic alteration found in premalignant stages of pancreatic adenocarcinoma is K-ras oncogene point mutation. The mechanistic basis for the inability of K-ras mutation to transform pancreatic ductal cells is unclear, although cooperating events with p16 inactivation, p53 mutation, and SMAD 4 mutation are recognized to be necessary. We have generated a novel mouse model in which the cytokeratin 19 promoter, specifically active in pancreatic ductal cells but not other cell types of the pancreas, is fused to mutant K-ras. This is of direct relevance to human pancreatic cancer because premalignant lesions are found specifically in ductal cells. There is dramatic periductal lymphocytic infiltration in the pancreata of transgenic mice, predominantly CD4+ T lymphocytes, which may act as an adaptive immune response to activated ras mediated signaling. In addition, gene array analysis reveals an induction of N cadherin in transgenic mice pancreatic ductal cells, the significance of which relates to promotion of cell adhesion and deterrence of cell migration. Apart from these important biological considerations, there is parallel activity of the cytokeratin 19 promoter in the stem cell region of the gastric epithelium, namely in mucous neck cells. Activated K-ras in this context causes mucous neck cell hyperplasia, a precursor to gastric adenocarcinoma. There is concomitant parietal cell decrease, which is a key step toward gastric adenocarcinoma. Taken together, we have defined how mutant K-ras signaling modulates important molecular events in the initiating events of pancreatic and gastric carcinogenesis. PMID- 12727810 TI - Transcription profile of Kaposi's sarcoma-associated herpesvirus in primary Kaposi's sarcoma lesions as determined by real-time PCR arrays. AB - Kaposi's sarcoma (KS) is the signature pathology of AIDS. KS-associated herpesvirus (KSHV/HHV-8) is causally linked to KS. Here, we report the first complete profile of KSHV transcription in primary KS lesions using a novel, real time PCR array. The KSHV latency I mRNAs [latency-associated nuclear antigen (LANA)/orf73, v-cyclin/orf72, and v-FLIP/orf71] were invariably present in all biopsies. Yet, viral lytic mRNAs were detectable in only a subset of tumors. Interestingly, there was a difference in the expression pattern of the viral IFN regulatory factors (vIRFs) encoded by KSHV. The vIRF-1/K9 clustered with LANA in KS, in contrast to its homologue, vIRF-3/LANA-2, which is transcribed only in KSHV-associated lymphomas. This suggests that the various vIRFs encoded by KSHV are important for KSHV latency as well as KS tumorigenesis and that their redundancy may be explained in part by a tissue-specific regulation. Clinical KSHV transcriptional profiling as described here will prove useful for the identification of KS tumor markers for diagnosis and as potential drug targets. Stratification by KSHV lytic transcription can identify lesions with a high percentage of lytically infected cells, which may respond to antiviral drugs. PMID- 12727811 TI - Preinvasive pancreatic neoplasia of ductal phenotype induced by acinar cell targeting of mutant Kras in transgenic mice. AB - Activating mutation of the Kras oncogene is the most frequent and perhaps the earliest genetic alteration associated with pancreatic cancer. To examine the link between mutant Kras and exocrine pancreatic cancer, we generated transgenic mice carrying an elastase-mutant Kras transgene, which targets expression to pancreatic acinar cells. Most elastase-Kras founder mice displayed perinatal pancreatic acinar cell hyperplasia and dysplasia. However, adult mice in two surviving lineages displayed preinvasive pancreatic neoplastic lesions with ductal morphology, thereby providing a unique mouse model in which lesion histotype and initiating genetic alteration overlap with the human disease. Our findings suggest that Kras mutation is associated with development of early stage duct-like lesions in pancreas, but that additional alterations must accompany progression to malignancy. PMID- 12727812 TI - Obtustatin: a potent selective inhibitor of alpha1beta1 integrin in vitro and angiogenesis in vivo. AB - A novel disintegrin, obtustatin, was purified from the venom of the Vipera lebetina obtusa viper. Obtustatin is the shortest disintegrin yet described, containing only 41 amino acids. It contains a similar pattern of cysteines to the short disintegrins echistatin and eristostatin but contains the sequence KTS rather than RGD in its active site loop. Obtustatin is a potent and selective inhibitor of alpha1beta1 integrin. It does not inhibit the closely related integrin alpha2beta1, nor a panel of other integrins tested. It does not inhibit ligand binding to the recombinant alpha1 I-domain. Importantly, obtustatin potently inhibited angiogenesis in vivo in the chicken chorioallantoic membrane assay, and in the Lewis lung syngeneic mouse model, it reduced tumor development by half, confirming and extending previous results on the relevance of alpha1beta1 integrin to angiogenesis and suggesting novel approaches to the generation of angiogenesis inhibitors. PMID- 12727813 TI - Lack of HIN-1 methylation in BRCA1-linked and "BRCA1-like" breast tumors. AB - We recently identified a candidate tumor suppressor gene, HIN-1, that is silenced due to methylation in the majority of sporadic breast carcinomas and is localized to 5q33-qter, an area frequently lost in BRCA1 tumors and thought to harbor a BRCA1 modifier gene. To establish whether germ-line mutations in HIN-1 may influence breast cancer risk, we sequenced the HIN-1 coding region in 10 familial breast cancer patients with positive logarithm of the odds scores of at least one of the markers flanking HIN-1. We also sequenced the HIN-1 coding region in 15 BRCA1 and 35 sporadic breast tumors to determine whether HIN-1 is the target of the frequent 5q loss in BRCA1 tumors. No sequence alterations were found in any of the cases analyzed. However, analysis of HIN-1 promoter methylation status revealed that in striking contrast to sporadic cases, there is a nearly complete lack of HIN-1 methylation in BRCA1 tumors (P < 0.0001). Sporadic breast tumors with a "BRCA1-like" histopathological phenotype also demonstrated significantly lower frequency of HIN-1 promoter methylation (P = 0.01) compared with other cancer types, and there was also a difference among tumors based on their estrogen receptor and HER2 status (P = 0.006), suggesting that HIN-1 methylation patterns are associated with specific breast cancer subtypes. PMID- 12727814 TI - Molecular characterization of circulating EBV DNA in the plasma of nasopharyngeal carcinoma and lymphoma patients. AB - Despite the increasing clinical applications of circulating EBV DNA analysis as a tumor marker, the molecular nature of these EBV DNA molecules remains unclear. We subjected plasma/serum samples of nasopharyngeal carcinoma and lymphoma patients to DNase digestion and ultracentrifugation and showed that circulating EBV DNA molecules are "naked" DNA fragments instead of being contained inside virions. We further showed that these EBV DNA fragments were relatively short, and 87% of them were shorter than 181 bp. These results provide fundamental information that may improve our understanding of the release of tumor-derived nucleic acids into the blood of cancer patients. PMID- 12727815 TI - CDKN1A and CDKN1B polymorphisms and risk of advanced prostate carcinoma. AB - A multigenic model of prostate cancer susceptibility has been proposed, in which common polymorphic variants of genes, such as the androgen and vitamin D receptor, contribute to tumorigenesis. The discovery of additional genetic factors that contribute to prostate cancer risk should provide opportunities for new approaches to the detection and treatment of this common malignancy. Herein, we examined single nucleotide polymorphic variants in the 3'-untranslated region of CDKN1A (p21(cip1)) and in codon 109 of CDKN1B (p27(kip1)) for association with advanced prostate cancer in a European-American population. Ninety-six cases and 106 controls were analyzed using PCR amplification and restriction digestion assays. CDKN1A genotype was scored as CC, CT, and TT on the basis of the digestion products. The CDKN1A genotypes CT and TT were associated with an increased risk of advanced prostate carcinoma compared with the CC genotype [odds ratio (OR), 2.24; 95% confidence interval (CI), 1.02-4.95]. The CDKN1B genotype was scored as VV, VG, or GG, again on the basis of the digestion products. The CDKN1B genotype VV was also associated with an increased risk of advanced prostate carcinoma (OR, 1.95; 95% CI, 1.09-3.47). These associations were particularly strong in those patients with androgen-independent disease [OR = 2.88 (95% CI, 1.19-6.97) and 2.11 (95% CI, 1.05-4.22) for high-risk genotypes of CDKN1A and CDKN1B, respectively]. In addition, the association of CDKN1B was particularly strong in the cohort of patients under the median age of diagnosis (OR, 2.23; 95% CI, 1.08-4.59). These results suggest that in a European-American population, CDKN1A and CDKN1B variants are associated with advanced prostate cancer. Analysis of CDKN1A and/or CDKN1B genotypes may prove useful in determining which patients are at risk for developing advanced prostate carcinoma and therefore would gain the most from aggressive screening, prophylaxis, and/or treatment. PMID- 12727816 TI - Bradykinin receptor subtype 1 expression and function in prostate cancer. AB - Kinins exert multiple pathophysiological functions, including vascular permeability and mitogenesis, by activating their cognate receptors, bradykinin subtype 1 receptor (B1R) and bradykinin subtype 2 receptor (B2R), which belong to the superfamily of G protein-coupled receptors. Tissue-specific expression pattern or contribution of the individual kinin receptors to pathological prostate cell growth is not known. We report here the differential expression of B1R and B2R in human benign and malignant prostate specimens. Whereas B2R is ubiquitously expressed, the B1R is detected only in prostatic intraepithelial neoplasia and malignant lesions and not in benign prostate tissues. Using androgen-insensitive prostate cancer PC3 cells, we show that specific stimulation of endogenous B1R promotes cell growth, migration, and invasion. These findings identify B1R as an early marker for pathological growth of the prostate and suggest its potential utility as a drug target effective for the treatment of prostate cancer. PMID- 12727817 TI - Site-directed mutations in the tumor-associated cytokine, autotaxin, eliminate nucleotide phosphodiesterase, lysophospholipase D, and motogenic activities. AB - The exo-enzyme autotaxin/NPP2 (ATX/NPP2) is a potent stimulator of cell migration, invasion, metastasis, and angiogenesis. Recently, ATX/NPP2 was found to possess lysophospholipase D (lyso-LPD) activity, generating the bioactive mediator lysophosphatidic acid from precursors. In the present study, we used site-directed mutagenesis to delineate the active domain of lysophospholipid catalytic activity and to examine potential overlap with the nucleotide phosphodiesterase domain. We found four amino acid residues obligatory for the phosphodiesterase, lyso-PLD, and migration-stimulating activities of ATX/NPP2, suggesting that 5'-nucleotide phosphodiesterase (PDE) and lyso-PLD share a common reaction mechanism and inviting design of enzymatic inhibitors as therapeutic agents for neoplastic disease. PMID- 12727818 TI - The cis decoy against the estrogen response element suppresses breast cancer cells via target disrupting c-fos not mitogen-activated protein kinase activity. AB - Breast cancer, the most common malignancy in women, has been demonstrated to be associated with the steroid hormone estrogen and its receptor (ER), a ligand activated transcription factor. Therefore, we developed a phosphorothiolate cis element decoy against the estrogen response element (ERE decoy) to target disruption of ER DNA binding and transcriptional activity. Here, we showed that the ERE decoy potently ablated the 17beta-estrogen-inducible cell proliferation and induced apoptosis of human breast carcinoma cells by functionally affecting expression of c-fos gene and AP-1 luciferase gene reporter activity. Specificity of the decoy was demonstrated by its ability to directly block ER binding to a cis-element probe and transactivation. Moreover, the decoy failed to inhibit ER mediated mitogen-activated protein kinase signaling pathways and cell growth of ER-negative breast cancer cells. Taken together, these data suggest that estrogen mediated cell growth of breast cancer cells can be preferentially restricted via targeted disruption of ER at the level of DNA binding by a novel and specific decoy strategy applied to steroid nuclear receptors. PMID- 12727819 TI - Cysteine-rich fibroblast growth factor receptor 1, a new marker for precancerous epithelial lesions defined by the human monoclonal antibody PAM-1. AB - Precancerous epithelial lesions are sites of uncontrolled cellular proliferation, generated by irreversible genetic changes. Not all of these lesions progress to invasive cancer, some may even regress, but early detection of abnormal cells can be crucial for survival of the patient. Diagnosis is mainly performed by using morphological parameters. Proliferation markers can facilitate the analysis, if they show a consistent expression, and distinguish between healthy and malignant cells. The fully human monoclonal IgM antibody PAM-1 was isolated from a patient with stomach carcinoma and binds to a new variant of cysteine-rich fibroblast growth factor receptor 1 (CFR-1). This CFR-1/PAM-1 receptor is expressed on nearly all of the epithelial cancers of every type and origin, but not on healthy tissue. It is also present on precursor lesions found in: Helicobacter pylori induced gastritis, intestinal metaplasia and dysplasia of the stomach, ulcerative colitis-related dysplasia and adenomas of the colon, Barrett's metaplasia and dysplasia of the esophagus, squamous cell metaplasia and dysplasia of the lung, and cervical intraepithelial neoplasia. The unique, growth-dependent expression of this new CFR-1 isoform makes the PAM-1 antibody an ideal diagnostic tool for the detection of precancerous and cancerous lesions. PMID- 12727820 TI - DNA mismatch repair deficiency stimulates N-ethyl-N-nitrosourea-induced mutagenesis and lymphomagenesis. AB - The primary role of the mismatch repair (MMR) system is the avoidance of mutations caused by replication and recombination errors. Furthermore, the lethality of methylating agents has been attributed to the processing of O(6) methylguanine lesions in DNA by MMR. Loss of the MSH2 protein completely abolishes repair function and results in reduced cell killing by methylating agents and accelerated accumulation of methylation-damage-induced mutations. This has raised the question as to whether MMR is also involved in the cellular response to other genotoxic insults. Here we describe that in mice deficient for Msh2, lymphomagenesis was strongly accelerated by an ethylating agent, N-ethyl-N nitrosourea (ENU), given at a dose that did not induce lymphomas in wild-type mice. This suggests that MMR deficiency and ENU-induced mutagenesis synergistically collaborate in inducing tumorigenesis. To study the interaction between MMR and ENU-induced DNA damage, we compared the lethality and mutagenicity of ENU in MSH2-proficient and -deficient mouse embryonic stem cells. Although MSH2-deficiency only slightly reduced the lethality of ENU, it strongly enhanced the mutagenicity of ENU. Mutation analysis of ENU-induced Hprt mutants revealed that base substitutions occurred predominantly at A-T base-pairs. These results suggest that MMR modulates the processing of ethylation damage at AT base pairs. PMID- 12727821 TI - Chronic lymphocytic leukemia with prostate infiltration mediated by specific clonal membrane-bound IgM. AB - Chronic lymphocytic leukemia (CLL) is characterized by an accumulation of monoclonal B lymphocytes in the hematopoietic organs. Rarely, CLL cells accumulate in a single atypical site. The mechanism underlying this unusual distribution of CLL cells has not been studied previously. We obtained peripheral blood from five patients having early stage CLL with heavy prostate infiltration. These patients' circulating CLL cells bound strongly in vitro to cultured prostate cell lines PC3, LNCaP, and DU145 and to short-term cultures of fresh prostate cells but not to colon, breast, or bladder cells. CLL cells from patients without prostate infiltration did not bind in vitro to any cell line. Peripheral blood CLL cells from one patient with CLL with heavy prostate infiltration were fused with a mouse-human heteromyeloma line to make hybridomas expressing the same monoclonal IgM as the patient's CLL cells. The hybridoma cells bound specifically to prostate cells. IgM secreted by the hybridoma blocked binding of the patient's CLL cells to prostate cells. Flow cytometry and immunohistochemistry demonstrated that the secreted IgM bound specifically to prostate cells. These results indicate that CLL with atypical prostate infiltration can be mediated by specific surface-bound IgM against an antigen expressed specifically by prostate cells and suggest that a similar mechanism might also apply to cases of CLL with atypical infiltration into other organs. PMID- 12727822 TI - Prostate secretory protein PSP-94 decreases tumor growth and hypercalcemia of malignancy in a syngenic in vivo model of prostate cancer. AB - Prostate cancer is a common malignancy affecting men, which is often associated with skeletal metastases resulting in significant morbidity and mortality. In this hormone-dependent cancer, low levels of a prostate secretory protein of 94 amino acids (PSP-94) are associated with advanced disease stage. In the current study, we have examined the effect of PSP-94 on prostate cancer growth and experimental metastases to the skeleton. For these studies, MatLyLu rat prostate cancer cells were transfected with full-length cDNA encoding parathyroid hormone related protein [PTHrP (MatLyLu-PTHrP cells)], which is known to be the major pathogenetic factor for malignancy-associated hypercalcemia. MatLyLu-PTHrP cells were inoculated s.c. into the right flank or via intracardiac route into the left ventricle of syngeneic male Copenhagen rats. Intracardiac inoculation of MatLyLu cells routinely results in the development of tumors in the lumbar vertebrae, resulting in hind-limb paralysis. Animals were infused with different doses of PSP-94 (0.1, 1.0, and 10.0 micro g/kg/day) starting on the day of tumor cell inoculation. Time of hind-limb paralysis and tumor volume were determined, and comparison was made between PSP-94-treated animals and control animals receiving vehicle alone. At the end of the study, animals were sacrificed, and plasma calcium, plasma PTHrP, and tumor PTHrP levels were determined. Whereas the highest dose of PSP-94 caused a modest but statistically significant delay in the development of hind-limb paralysis, a marked dose-dependent decrease in primary tumor volume was seen in experimental animals receiving PSP-94 due to its ability to promote tumor cell apoptosis. Furthermore, whereas control animals routinely developed hypercalcemia due to PTHrP production, treatment with PSP-94 led to a near normalization of plasma calcium and a marked reduction in PTHrP production as determined by radioimmunoassay and immunohistochemistry. Collectively, these results demonstrate the ability of PSP-94 to be an effective treatment modality for prostate cancer, where decrease in plasma PTHrP and calcium levels can serve as useful biochemical markers for monitoring the efficacy of this novel antitumor agent. PMID- 12727823 TI - Alphavbeta3 integrin antagonist S247 decreases colon cancer metastasis and angiogenesis and improves survival in mice. AB - Members of the integrin family influence several aspects of tumor progression and metastasis, including cell survival, proliferation, and angiogenesis. Specific integrins such as alpha(v)beta(3) and alpha(v)beta(5) are involved in regulating endothelial cell function, and thus angiogenesis. We evaluated the effect of the alpha(v)beta(3)/alpha(v)beta(5) integrin antagonist S247 on the growth and angiogenesis of colon cancer liver metastases in an orthotopic murine model. Murine colon cancer cells were injected into the spleens of BALB/c mice to produce liver metastases. On day 7, miniature osmotic pumps were implanted into the subcutis to continuously infuse either saline or 70 mg/kg/day S247. All mice were sacrificed when control mice became moribund. Mice that received S247 developed significantly fewer liver metastases than did controls (P < 0.05). Using the same model, a subsequent survival study was performed. Mice were sacrificed when moribund as determined by an observer blinded to the treatment given. Treatment with S247 significantly prolonged overall survival (P < 0.05). Interestingly, primary tumors in the spleen were the cause of death in the S247 treated group as S247 appeared to have little effect on these tumors. Immunohistochemical staining demonstrated a significant reduction of vessels in liver metastases of S247-treated mice (P < 0.001), a significant increase in endothelial cell apoptosis (P < 0.05), and a significant decrease in pericyte coverage (P < 0.0001). To determine the role of S247 on angiogenesis, we examined the effect of S247 in vitro on human umbilical vein endothelial cells (HUVECs) and human vascular smooth muscle cells (hVSMCs). The addition of S247 to HUVECs and hVSMCs growing on vitronectin-coated flasks and in Matrigel significantly impaired cell growth and colony formation, respectively (P < 0.05). Furthermore, S247 completely inhibited the attachment of HUVECs and hVSMCs and increased apoptosis by six- to 9fold compared with controls. In in vitro invasion assays, S247-treated cells demonstrated decreased migration (P < 0.05). In conclusion, S247 demonstrated significant antimetastatic and antiangiogenic activity and impaired both endothelial and hVSMC/pericyte function in vitro and in vivo. The use of agents such as integrin antagonists that target multiple cell types involved in angiogenesis may be a more effective method of inhibiting angiogenesis than agents targeting only the endothelial cells. PMID- 12727824 TI - Inhibition of the Raf/MEK/ERK pathway up-regulates expression of the coxsackievirus and adenovirus receptor in cancer cells. AB - Recombinant adenoviruses are presently being tested clinically as a new strategy for the treatment of cancer. An important determining factor for the successful entry of such adenoviruses into target cells is expression of the coxsackievirus and adenovirus receptor (CAR) at the cell surface. Recent observations suggest that expression of this receptor, which physiologically participates in formation of cell-cell adhesions, is frequently reduced in highly malignant cancer cells. This raises the possibility that those tumors representing the greatest therapeutic challenge might be the least susceptible to infection with therapeutic adenoviruses. We explored the role of the Raf-MEK-ERK pathway on CAR expression in a panel of cancer cells because this pathway is frequently up regulated in cancer cells and is known to down-regulate cell-cell adhesion molecules. We found that disruption of signaling through the Raf-MEK-ERK pathway by inhibition of MEK up-regulated CAR expression, which was accompanied by increased representation of the protein at the cell surface. After Raf-MEK-ERK inhibition, adenovirus entry into cells was increased and cell killing by replication competent adenoviruses was enhanced in a CAR-dependent manner. Conversely, induction of Raf-1 resulted in reduction and disruption of CAR expression at the cell surface. We conclude that loss of CAR expression in cancer cells is, at least in part, mediated through the Raf-MEK-ERK signal transduction pathway and that pharmacological restoration of CAR at the cell surface could improve adenovirus-based treatments of cancer. PMID- 12727825 TI - Osteoprotegerin/osteoclastogenesis inhibitory factor decreases human prostate cancer burden in human adult bone implanted into nonobese diabetic/severe combined immunodeficient mice. AB - Human prostate cancer frequently metastasizes to bone, where it gives rise to osteoblastic bone metastases with an underlying osteoclastic component and subsequent bone pain. However, the importance of osteoclastogenesis in the development of prostate cancer bone lesions in humans is unclear. Osteoprotegerin/osteoclastogenesis inhibitory factor (OCIF) is a member of the tumor necrosis factor receptor family and a novel secreted protein, and it is a negative regulator of osteoclast differentiation, activation, and survival both in vitro and in vivo. In the present study we used a model in which human LNCaP prostate cancer cells that give rise to osteoblastic bone tumors were injected directly into the intramedullary space of human adult bone implanted into nonobese diabetic/severe combined immunodeficient mice to investigate whether the new bone-resorption inhibitor osteoprotegerin/OCIF would inhibit the development of new bone tumors and the progression of established osteoblastic bone tumors. The mice were given consecutive daily s.c. injections of recombinant human OCIF (rhOCIF; 100 micro g/mouse/day) for 2 weeks starting either immediately or 2 weeks after injection of the LNCaP cells. In both protocols, rhOCIF markedly inhibited both the development of bone tumors and the progression of established bone tumor foci quantified by histological examination. Histomorphometrical analysis revealed that rhOCIF markedly reduced the number of osteoclasts and the size of the tumors at the bone sites, but that it had no effect on the local growth of s.c. LNCaP tumors or on LNCaP cell proliferation in culture. These findings demonstrate that osteoclasts play an important role in bone tumor by prostate cancer, and that rhOCIF decreases the LNCaP prostate cancer burden selectively in bone, suppresses the progression of established tumor lesions, and prevents the development of new lesions. These results suggest that inhibition of osteoclastic bone resorption may be an effective therapy for the treatment of prostate cancer that has colonized bone. PMID- 12727826 TI - Induction of autophagic cell death in malignant glioma cells by arsenic trioxide. AB - Recent clinical data shows that arsenic trioxide (As(2)O(3)) causes remission in patients with acute promyelocytic leukemia and multiple myeloma without severe side effects. Laboratory data suggest that As(2)O(3) induces apoptosis or cell differentiation of hematopoietic or solid tumor cells. To date, there has been no study on the effects of As(2)O(3) on glioma cells. In this study, we investigated the in vitro effect of As(2)O(3) on cell growth inhibition and cell death mechanisms in human glioma cells. As(2)O(3) significantly inhibited the proliferation of all six of the glioma cell lines (U373, U87, U251, GB1, A-172, and T98G) tested in this study in a dose-dependent manner. The IC(50) of As(2)O(3) for all of the tumor cell lines was <2 micro M. Previous studies have shown that this is a clinically safe concentration. Treatment with 2 micro M As(2)O(3) induced G(2)/M arrest in all of the glioma cell lines. Autophagy (programmed cell death type II), but not apoptosis (programmed cell death type I), was detected by electron microscopy in U-373-MG cells treated with 2 micro M As(2)O(3). Caspase inhibitors did not halt As(2)O(3)-induced cell death. Furthermore, combination of As(2)O(3) with bafilomycin A1 autophagy inhibitor enhanced the antitumor effect of As(2)O(3) through induction of apoptosis. These findings suggest that As(2)O(3) at a clinically safe concentration may be an effective chemotherapeutic agent for malignant gliomas. PMID- 12727827 TI - Redox regulation of the G1 to S phase transition in the mouse embryo fibroblast cell cycle. AB - The hypothesis that intracellular oxidation/reduction (redox) reactions regulate the G(0)-G(1) to S-phase transition in the mouse embryonic fibroblast cell cycle was investigated. Intracellular redox state was modulated with a thiol antioxidant, N-acetyl-L-cysteine (NAC), and cell cycle progression was measured using BrdUrd pulse-chase and flow cytometric analysis. Treatment with NAC for 12 h resulted in an approximately 6-fold increase in intracellular low-molecular weight thiols and a decrease in the MFI of an oxidation-sensitive probe, dihydrofluorescein diacetate, indicating a shift in the intracellular redox state toward a more reducing environment. NAC-induced alterations in redox state caused selective delays in progression from G(0)-G(1) to S phase in serum-starved cells that were serum stimulated to reenter the cell cycle as well as to inhibit progression from G(1) to S phase in asynchronous cultures with no significant alterations in S phase, and G(2)+M transits. NAC treatment also showed a 70% decrease in cyclin D1 protein levels and a 3-4-fold increase in p27 protein levels, which correlated with decreased retinoblastoma protein phosphorylation. Cells released from the NAC treatment showed a transient increase in dihydrofluorescein fluorescence and oxidized glutathione content between 0 and 8 h after release, indicating a shift in intracellular redox state to a more oxidizing environment. These changes in redox state were followed by an increase in cyclin D1, a decrease in p27, retinoblastoma protein hyperphosphorylation and subsequent entry into S phase by 8-12 h after the removal of NAC. These results support the hypothesis that a redox cycle within the mammalian cell cycle might provide a mechanistic link between the metabolic processes early in G(1) and the activation of G(1)-regulatory proteins in preparation for the entry of cells into S phase. PMID- 12727828 TI - Histone deacetylase inhibitors promote STI571-mediated apoptosis in STI571 sensitive and -resistant Bcr/Abl+ human myeloid leukemia cells. AB - Interactions between the Bcr/Abl kinase inhibitor STI571 (Gleevec, imatinib mesylate) and histone deacetylase inhibitors (HDIs) have been examined in STI571 sensitive and -resistant Bcr/Abl(+) human leukemia cells (K562 and LAMA 84). Cotreatment of K562 cells with 250 nM imatinib mesylate and 2.0 micro M suberoylanilide hydroxamic acid (SAHA) for 24 h, exposures that were minimally toxic alone, resulted in a marked increase in mitochondrial damage (e.g., cytochrome c, Smac/DIABLO, and apoptosis-inducing factor release), caspase activation, and apoptosis. Similar events were observed in other Bcr/Abl(+) cells (i.e., LAMA 84), and in cells exposed to STI571 in combination with the HDI sodium butyrate. Coexposure of cells to HDIs in conjunction with STI571 resulted in multiple perturbations in signaling and cell cycle-regulatory proteins, including down-regulation of Raf, phospho-mitogen-activated protein kinase kinase (MEK), phospho-extracellular signal-regulated kinase (ERK), phospho-Akt, phospho signal transducers and activators of transcription 5, cyclin D1, and Mcl-1, accompanied by dephosphorylation and cleavage of retinoblastoma protein and a striking increase in phosphorylation of c-Jun NH(2)-terminal kinase. Coexposure of Bcr/Abl(+) cells to STI571 also blocked SAHA-mediated induction of p21(CIP1) and resulted in down-regulation of Bcr/Abl protein expression. STI571 and SAHA also interacted synergistically to induce apoptosis in STI571-resistant K562 and LAMA 84 cells that display increased Bcr/Abl protein expression. Lastly, inducible expression of a constitutively active MEK1/2 construct significantly attenuated SAHA/STI571-mediated apoptosis in K562 cells, implicating disruption of the Raf/MEK/ERK axis in synergistic antileukemic effects of this drug combination. Together, these findings indicate that combined exposure of Bcr/Abl(+) cells to the kinase inhibitor STI571 and HDIs leads to diverse perturbations in signaling and cell cycle-regulatory proteins, associated with a marked increase in mitochondrial damage and cell death. They also raise the possibility that this strategy may be effective in some Bcr/Abl(+) cells that are resistant to STI571 through increased Bcr/Abl expression. PMID- 12727829 TI - Immunological and clinical responses in metastatic renal cancer patients vaccinated with tumor RNA-transfected dendritic cells. AB - Autologous dendritic cells transfected with total renal tumor RNA have been shown to be potent stimulators of CTLs and antitumor immunity in vitro. A Phase I trial was conducted to evaluate this strategy for feasibility, safety, and efficacy to induce tumor-specific T-cell responses in subjects with metastatic renal cell carcinoma. Renal tumor RNA-transfected dendritic cells were administered to 10 evaluable study patients with no evidence of dose-limiting toxicity or vaccine related adverse effects including autoimmunity. In six of seven evaluable subjects, expansion of tumor-specific T cells was detected after immunization. The vaccine-induced T-cell reactivities were directed against a broad set of renal tumor-associated antigens, including telomerase reverse transcriptase, G250, and oncofetal antigen, but not against self-antigens expressed by normal renal tissues. Although most patients underwent secondary therapies after vaccination, tumor-related mortality of the study subjects was unexpectedly low with only 3 of 10 patients dying from disease after a mean follow-up of 19.8 months. These data provide a scientific rationale for continued clinical investigation of this polyvalent vaccine strategy in the treatment of metastatic renal cell carcinoma and, potentially, other cancers. PMID- 12727830 TI - Repopulation in murine breast tumors during and after sequential treatments with cyclophosphamide and 5-fluorouracil. AB - The rate of repopulation of surviving cells increases during fractionated radiotherapy and limits the ability to control tumors. Repopulation is likely to be more important during the longer intervals between courses of chemotherapy and is a potential cause of clinical resistance to chemotherapy. To evaluate the rate of repopulation of surviving cells in MXT and EMT-6 mouse breast tumors between successive cycles of chemotherapy, tumor-bearing mice were treated with up to three weekly cycles of cyclophosphamide or 5-fluorouracil (5FU). Animals were killed at different intervals during and after chemotherapy, and uptake of 5'bromodeoxyuridine and immunohistochemical staining were selected to study repopulation. When tumors regrew after a single treatment, the proliferative rate of tumor cells returned to control values. During successive courses of chemotherapy, the proliferative rate of surviving cells increased in both tumors, e.g., in 5FU-treated MXT tumors, the percentages of 5'bromodeoxyuridine-labeled proliferating cells at 7 days after the first, second, and third treatment were 25 +/- 12%, 29 +/- 9%, and 27 +/- 11%; it was 32 +/- 6% at 14 days after the third treatment. The corresponding value in control tumors was 19 +/- 2% (P < 0.05 for each comparison). Accelerating repopulation after sequential treatments with cyclophosphamide or 5FU can lead to effective drug resistance in the absence of changes in the sensitivity of constituent cells. PMID- 12727831 TI - Inhibition of heat shock protein 90 function down-regulates Akt kinase and sensitizes tumors to Taxol. AB - The phosphatidylinositol 3'-kinase/Akt pathway is activated frequently in human cancer, and has been implicated in tumor proliferation, cell survival, and resistance to apoptotic stimuli. Akt forms a complex with heat shock protein (Hsp) 90 and Cdc37, and inhibitors of Hsp90 cause Akt degradation. 17-allylamino 17-demethoxygeldanamycin (17-AGG) is an Hsp90 inhibitor currently in Phase I clinical trial. 17-AAG inhibits Akt activation and expression in tumors, and has antitumor activity in breast cancer xenografts. The combination of 17-AAG and Taxol is synergistic, and 17-AAG sensitizes tumor cells to Taxol-induced apoptosis in a schedule-dependent manner. Transfection of membrane-bound p110 PI3k prevented 17-AAG inactivation of Akt and abrogated the enhancement of Taxol induced apoptosis caused by the drug. 17-AAG and Taxol could be administered together at their maximally tolerated doses to tumor-bearing mice. Doses of 17 AAG that induce HER2 degradation and cause Akt inactivation but have no single agent activity were effective in sensitizing tumors to Taxol. Enhancement was schedule-dependent and maximal when Taxol and 17-AAG were administered on the same day. These results suggest that Hsp90 inhibitors can effectively suppress Akt activity in animal models of human cancer at nontoxic doses, thus sensitizing tumor cells to proapoptotic stimuli. PMID- 12727832 TI - Intradermal injection, as opposed to subcutaneous injection, enhances immunogenicity and suppresses tumorigenicity of tumor cells. AB - Tumor cell immunogenicity depends heavily upon the microenvironment in which the cells grow. We have compared the tumorigenicity and immunogenicity of the same tumor cells when injected either into the dermis, a tissue containing numerous dendritic cells (DCs), or s.c., at a site which contains only few DCs. After s.c. injection, progressive tumors were constantly obtained, whereas most intradermal injections did not give rise to tumor and immunized animals against additional challenge. We present evidence that the high density of DCs at dermal sites facilitates the capture of tumor antigens and that local inflammation induces maturation of the DCs and their migration into draining lymph nodes. PMID- 12727833 TI - Tumor-induced interleukin 10 suppresses the ability of splenic dendritic cells to stimulate CD4 and CD8 T-cell responses. AB - Dendritic cell (DC) maturation and function are influenced by the surrounding cytokine milieu. We demonstrate tumor-associated suppression of DCs in stimulating allogeneic and tumor-specific CTL and type 1 (IFN-gamma-producing) responses in both CD4- and CD8-positive T cells. DCs from MB49-bearing female mice fail to stimulate proliferative and IFN-gamma-producing responses in allogeneic mixed lymphocyte cultures. MB49 also inhibited DC function in stimulating type 1 responses against our tumor-specific antigen, the male antigen, HY. DCs from MB49-bearing male mice were unable to restimulate effective HY-specific CTLs or IFN-gamma. Tumor-induced interleukin (IL) 10 was found to be specifically responsible for DC dysfunction in response to antigenic driven maturation. This was demonstrated by restoration of DC function in splenic DCs from MB49-bearing female IL-10 knockout mice (HY disparity), whereas not in MB49 bearing male IL-10 knockout mice (no HY disparity). Finally, any tumor-induced systemic inhibitory effect on bone marrow precursors could be overcome by generation of bone marrow-derived DCs ex vivo. These bone marrow-derived DCs derived from MB49-bearing B6 mice were capable of inducing control levels of proliferation in allogeneic mixed lymphocyte reactions and a type 1 (IFN-gamma) cytokine profile. The BM-DCs were also capable of restimulating HY-specific CTL and IFN-gamma production. These studies reveal the tumor-associated in vivo effects of IL-10 inhibition on DC function in eliciting a type 1 immune response in both allogeneic and tumor-specific responses. PMID- 12727834 TI - The cytotoxic T-lymphocyte response against a poorly immunogenic mammary adenocarcinoma is focused on a single immunodominant class I epitope derived from the gp70 Env product of an endogenous retrovirus. AB - The TS/A mouse mammary adenocarcinoma is a poorly immunogenic tumor widely used in preclinical models of cancer immunotherapy. CTLs have often been indicated as important in TS/A tumor destruction, but their generation in this model has been rarely studied, nor have their precise target(s) been identified. We hypothesized that the gp70 Env product of an endogenous murine leukemia virus could be a target antigen for TS/A-specific CTLs and investigated this possibility in four different TS/A cell lines engineered with the genes that encode IFN-alpha, IFN gamma, interleukin-4, and B7.1, respectively. All tumor cell lines expressed gp70, albeit at different levels, as demonstrated by reverse transcription-PCR analysis. Transfected tumor cells exhibited a delayed growth in vivo, and partial tumor regression. Spleen cells from mice that displayed tumor regression had high percentages of CD8(+) T cells that were specifically stained with L(d) tetramers loaded with gp70(423-431), the antigenic epitope of gp70 protein. Mixed leukocyte peptide and mixed leukocyte-tumor cultures, set up by stimulating splenocytes with the immunogenic peptide and with transfected TS/A tumor cells, respectively, resulted in similar large increases in tetramer-reactive CD8(+) T cells and showed high lytic activity specific for gp70(423-431). Finally, in a Cold Target Inhibition assay, lytic activity of a mixed leukocyte-tumor culture was inhibited in an overlapping fashion by both the TS/A line used for restimulation and 293L(d) cells loaded with gp70(423-431) peptide, but not by 293L(d) cells pulsed with an irrelevant H-2 L(d) epitope, thus demonstrating that all or most of the cytotoxic activity was directed exclusively against this antigenic epitope. PMID- 12727835 TI - Triple analysis of the cancer epigenome: an integrated microarray system for assessing gene expression, DNA methylation, and histone acetylation. AB - We developed a novel microarray system to assess gene expression, DNA methylation, and histone acetylation in parallel, and to dissect the complex hierarchy of epigenetic changes in cancer. An integrated microarray panel consisting of 1507 short CpG island tags located at the 5'-end regions (including the first exons) was used to assess effects of epigenetic treatments on a human epithelial ovarian cancer cell line. Treatment with methylation (5-aza-2' deoxycytidine) or deacetylation (trichostatin A) inhibitors alone resulted in up regulation of 1.9 or 1.1% of the genes analyzed; however, the combined treatment resulted in synergistic reactivation of more genes (10.4%; P < 0.001 versus either treatment alone). On the basis of either primary or secondary responses to the treatments, genes were identified as methylation-dependent or -independent. Synergistic reactivation of the methylation-dependent genes by 5-aza-2' deoxycytidine plus trichostatin A revealed a functional interaction between methylated promoters and deacetylated histones. Increased expression of some methylation-independent genes was associated with enhanced histone acetylation, but up-regulation of most of the genes identified using this technology was because of events downstream of the epigenetic cascade. We demonstrate proof of principle for using the triple microarray system in analyzing the dynamic relationship between transcription factors and promoter targets in cancer genomes. PMID- 12727837 TI - The Septin 9 (MSF) gene is amplified and overexpressed in mouse mammary gland adenocarcinomas and human breast cancer cell lines. AB - The expression of polyomavirus middle T antigen under the control of the mouse mammary tumor virus promoter in transgenic mice results in the induction of aggressive mammary gland adenocarcinomas at an early age. We screened 26 tumors for chromosomal aneuploidies using SKY and CGH. In 70% of the tumor samples we could detect high-level copy number gains, which mapped to chromosome band 11E2, a region orthologous to human 17q25.3. We then identified a bacterial artificial chromosome clone that labeled double-minute chromosomes found in the tumor metaphases. This bacterial artificial chromosome clone showed sequence homology to a member of the septin gene family. Real-time PCR analysis revealed a consistently increased expression of septin 9 (Sept9), not only in polyomavirus middle T antigen-induced, but in a wide variety of mouse models of breast cancer. Six of 9 human tumor cell lines also revealed elevated expression levels of Sept9. The family of septin genes is involved in a plethora of cellular processes, including cytokinesis in yeast and vesicle transport, and possesses GTPase activity. We identified down-regulation of Thsp1- and Bax-regulated apoptotic response in those tumors with Sept9 overexpression, an effect that could be reversed by inhibiting Sept9 expression using transfection with small interference RNA. Our results now suggest that signaling via members of the septin family plays a novel and common role in breast tumorigenesis. PMID- 12727836 TI - The protein kinase Akt induces epithelial mesenchymal transition and promotes enhanced motility and invasiveness of squamous cell carcinoma lines. AB - Epithelial-mesenchymal transition (EMT) is an important process during development and oncogenesis by which epithelial cells acquire fibroblast-like properties and show reduced intercellular adhesion and increased motility. Squamous cell carcinoma lines engineered to express constitutively active Akt underwent EMT, characterized by down-regulation of the epithelial markers desmoplakin, E-cadherin, and beta-catenin and up-regulation of the mesenchymal marker vimentin. The cells lost epithelial cell morphology and acquired fibroblast-like properties. Additionally, E-cadherin was down-regulated transcriptionally. The cells expressing constitutively active Akt exhibited reduced cell-cell adhesion, increased motility on fibronectin-coated surfaces, and increased invasiveness in animals. AKT is activated in many human carcinomas, and the AKT-driven EMT may confer the motility required for tissue invasion and metastasis. These findings suggest that future therapies based on AKT inhibition may complement conventional treatments by controlling tumor cell invasion and metastasis. PMID- 12727838 TI - Overexpression of high mobility group box 1 in gastrointestinal stromal tumors with KIT mutation. AB - Gain-of-function mutations of KIT are common genetic events in gastrointestinal stromal tumors (GISTs). To investigate the molecular characteristics of KIT mutations in GISTs, 20 GISTs (14 GISTs with KIT mutation and 6 GISTs without KIT mutation) were analyzed by two-dimensional electrophoresis and matrix-associated laser desorption ionization mass spectrophotometry-time of flight. Comparative analysis of the respective spot patterns on two-dimensional electrophoresis showed that HMGB1, an intranuclear protein that interacts with several transcription factors and plays a role in tumor metastasis after its secretion, was overexpressed in GISTs with KIT mutation. All of the 14 GISTs with KIT mutation, and only 2 of 6 GISTs without KIT mutation, revealed HMGB1 expression. Of the GISTs with KIT mutation, 12 (86%) showed strong expression of HMGB1, more than three times higher in intensity than the maximum observed in the 6 GISTs without KIT mutation by two-dimensional electrophoresis analysis. The overexpression of HMGB1 was further supported by Western blot analysis, and directly related to matrix metalloproteinase 2 overexpression. Our results indicate that the overexpression of HMGB1 is common in GISTs and is related to the KIT mutation, and that this may play a role in the tumorigenesis of GISTs because overexpressed HMGB1 could accelerate genes related to tumor growth and invasion. PMID- 12727839 TI - Gene expression profiling detects gene amplification and differentiates tumor types in breast cancer. AB - Global gene expression analysis using microarrays has been used to characterize the molecular profile of tumors. Gene expression variability at the mRNA level can be caused by a number of different events, including novel signaling, downstream activation of transcription enhancers or silencers, somatic mutation, and genetic amplification or deletion. Genomic amplifications are commonly observed in cancer and often include known oncogenes. The tyrosine kinase-type cell surface receptor, ERBB2, is an oncogene located on chromosome 17q21.1 that is amplified in 10-40% of breast tumors. We report for the first time that phenylethanolamine N-methyltransferase (PNMT), proteasome subunit, beta type 3 (PSMB3), ribosomal protein L19 (RPL19), and nuclear receptor subfamily 1, group D, member 1 (NR1D1) are coexpressed with ERBB2 in 34 breast cancer biopsies and also mapped within the same chromosomal location as the ERBB2 gene. Consistent with previous reports, we also observed that the steroidogenic acute regulatory protein-related gene, MLN64, and growth factor receptor bound protein 7 were coexpressed with ERBB2. Coexpression and colocalization of PNMT and MLN64 with ERBB2 suggested that the amplification of ERBB2 includes the chromosomal region harboring these genes. This hypothesis was validated in a subset of 12 biopsies. Gene amplification of ERBB2, PNMT, and MLN64 significantly correlated with increased mRNA gene expression (P < 0.05). These results suggest that gene expression profiling of breast biopsies may become a valuable method for adequately characterizing and choosing treatment modality for patients with breast cancer. PMID- 12727840 TI - Molecular description of evolving paclitaxel resistance in the SKOV-3 human ovarian carcinoma cell line. AB - Ovarian cancer is currently the most lethal gynecological malignancy in the United States. Although effective therapies exist, the acquisition of multidrug resistance within persisting tumor cells renders curative therapies elusive for the majority of women with ovarian cancer. In an attempt to better define the evolution of paclitaxel resistance, three SKOV-3 sublines were selected during successive rounds of exposure to increasing paclitaxel concentrations. The sublines were selected to represent early (0.003 micro M), intermediate (0.03 micro M), and late (0.3 micro M) paclitaxel resistance. RNA from these cell lines, SKOV-3(0.003TR), SKOV-3(0.03TR), and SKOV-3(0.3TR), as well as the parent cell line SKOV-3, was analyzed by cDNA array to evaluate transcript expression profiles. Arrays were performed using Affymetrix HG-U95Av2 arrays, which contain probes for approximately 9600 known human genes. Signal intensities were calculated by Microarray Suite 5.0 (Affymetrix, Santa Clara, CA). Expression patterns were analyzed by Affymetrix Data Mining Tool 3.0 with filtering of expression patterns for fold change in expression (maximum divided by minimum expression value/gene) and for variation of expression (maximum minus minimum expression value/gene). This analysis dismissed approximately 11,000 of approximately 12,000 expression patterns. The remaining approximately 1000 expression patterns were normalized and segregated into 20 partitions of a self organizing map (SOM). The resulting SOM discriminates between genes, which are differentially expressed in early versus intermediate versus late paclitaxel resistance. For example, multidrug resistance 1 transcript expression is not elevated in SKOV-3(0.003TR) as compared with parental SKOV-3 but demonstrates elevated expression in SKOV-3(0.03TR) and SKOV-3(0.3TR). In contrast, SOM analysis demonstrates early (SKOV-3(0.003TR)) transcriptional changes in a wide variety of genes, including gene families involved in cell growth/maintenance, cell structure, signal transduction, and inflammatory response. The use of array analysis with SOMs in sublines with progressive paclitaxel resistance can successfully define an evolution of resistance. Such an analysis may be useful at defining candidate gene families involved in the early-drug resistance phenotype. PMID- 12727842 TI - Genome-wide genetic characterization of bladder cancer: a comparison of high density single-nucleotide polymorphism arrays and PCR-based microsatellite analysis. AB - Most human cancers are characterized by genomic instability, the accumulation of multiple genetic alterations, and allelic imbalance throughout the genome. Loss of heterozygosity (LOH) is a common form of allelic imbalance, and the detection of LOH has been used to identify genomic regions that harbor tumor suppressor genes and to characterize different tumor types, pathological stages and progression. Global patterns of LOH can be discerned by allelotyping of tumors with polymorphic genetic markers. Microsatellites are reliable genetic markers for studying LOH, but typically only a modest number of microsatellites are tested in LOH studies because the genotyping procedure can be laborious. Here we describe the use of a new alternative approach to comprehensive allelotyping in which samples are genotyped for nearly 1500 single-nucleotide polymorphism (SNP) loci distributed across all human autosomal arms. We examined the pattern of allelic imbalance in human transitional cell carcinomas of the urinary bladder including 36 primary tumors and 1 recurrent tumor with matched normal DNAs. The call rate for all SNPs was 78.5 +/- 1.87% overall samples. Overall, the median number of allelic imbalance was 47.5, ranging from 20 to 118. The mean number of allelic imbalances was 36.58, 51.30, and 67.78 for pT(a), pT(1), and > or =pT(2), respectively, and also increased by grade. The SNP microarray analysis result was validated by comparison with microsatellite allelotype analysis of 118 markers in the same tumors. Overall, the two methods produced consistent loss patterns at informative loci. The SNP assay discovered previously undiscovered allelic imbalances at chromosomal arms 12q, 16p, 1p, and 2q. The detection of LOH and other chromosomal changes using large numbers of SNP markers should enable rapid and accurate identification of allelic imbalance patterns that will facilitate the mapping and identification of important cancer genes. Moreover, SNP analysis raises the possibility of individual tumor genome-wide allelotyping with potential prognostic and diagnostic applications. PMID- 12727843 TI - Parallel overexpression of seven kallikrein genes in ovarian cancer. AB - Recent evidence suggests that many members of the human kallikrein (KLK) gene family are differentially regulated in ovarian cancer and have potential as diagnostic and/or prognostic markers. We used the serial analysis of gene expression and expressed sequence tag databases of the Cancer Genome Anatomy Project to perform in silico analyses of the expression pattern of the 15 human KLK genes in normal and cancerous ovarian tissues and cell lines. We found that seven KLK genes (KLK5, KLK6, KLK7, KLK8, KLK10, KLK11, and KLK14) are up regulated in ovarian cancer. Probing 2 normal and 10 ovarian cancer serial analysis of gene expression libraries with gene-specific tags for each KLK indicated that whereas no expression was detected in any normal libraries (with the exception of KLK10 and KLK11), these KLKs were found to be expressed with moderate densities (103-408 tags per million) in 40-60% of the ovarian cancer libraries analyzed. These data were verified by screening the expressed sequence tag databases, where 78 of 79 mRNA clones isolated for these genes were from ovarian cancer libraries. X-profiler comparison of the pools of normal and cancerous ovaries identified a significant difference in expression levels for six of the seven KLKs. We experimentally verified the overexpression of six KLK proteins in cancer versus normal or benign tissues with highly sensitive and specific immunofluorometric assays. A statistically significant stepwise increase in protein levels was found among normal, benign, and cancerous ovarian tissues. The expression of five KLKs showed a strong degree of correlation at the protein level, suggesting the existence of a common mechanism or pathway that controls the expression of this group of adjacent genes during ovarian cancer progression. PMID- 12727841 TI - Constitutive activation of nuclear factor kappaB p50/p65 and Fra-1 and JunD is essential for deregulated interleukin 6 expression in prostate cancer. AB - To date, no effective treatment for patients with advanced androgen-independent prostate cancer is available, whereas androgen ablation therapy, surgery, and radiation therapy are effective in treating local, androgen-dependent tumors. The mechanisms underlying the differences between androgen-dependent and -independent prostate cancer remain elusive. Interleukin (IL)-6 is a pleiotropic cytokine whose expression under normal physiological conditions is tightly controlled. However, aberrant constitutive IL-6 gene expression has been implicated in prostate cancer progression and resistance to chemotherapy and has been directly linked to prostate cancer morbidity and mortality. Particularly striking is the large increase in the expression of IL-6 in hormone-refractory prostate cancer. IL-6, in addition to its role as an immunomodulatory cytokine, functions as a growth and differentiation factor for prostate cancer cells. To determine the molecular mechanisms that lead to deregulated IL-6 expression in advanced prostate cancer, we examined the regulatory elements involved in IL-6 gene expression in androgen-independent prostate cancer cells. We demonstrate that, in contrast to the androgen-sensitive LNCaP cells, androgen-insensitive PC-3 and DU145 cells express high levels of IL-6 protein and mRNA due to enhanced promoter activity. Deregulated activation of the IL-6 promoter is for the most part mediated by a combined constitutive activation of the nuclear factor (NF)-kappaB p50 and p65 and the activator protein 1 (AP-1) JunD and Fra-1 family members as demonstrated by electrophoretic mobility shift assays, site-directed mutagenesis, and transfection experiments. Mutation of the NF-kappaB and AP-1 sites drastically reduces IL-6 promoter activity in both androgen-independent prostate cancer cell lines. Additionally, inhibition of these transcription factors using adenovirus vectors encoding either the IkappaBalpha repressor gene or a dominant negative JunD mutant leads to a strong down-regulation of IL-6 gene expression at the mRNA and protein level as measured by real-time PCR and ELISA, respectively. Furthermore, the blockade of IL-6 gene expression results in drastic inhibition of the constitutively activated signal transducers and activators of transcription 3 signaling pathway in DU145 cells. Our data demonstrate for the first time that a combined aberrant activation of NF-kappaB p50 and p65 and AP-1 JunD and Fra-1 in androgen-independent prostate cancer cells results in deregulated IL-6 expression, suggesting a novel potential entry point for therapeutic intervention in prostate cancer. PMID- 12727844 TI - Androgen-independent growth of LNCaP prostate cancer cells is mediated by gain-of function mutant p53. AB - Mutations of p53 are common in hormone-refractory prostate cancer (CaP), suggesting the possibility that these mutations may be involved in the progression of CaP to androgen-independent (AI) growth. However, at present no direct evidence has been presented linking p53 mutations with AI growth of CaP. We established five stably transfected LNCaP cell lines: four containing gain-of function (GOF) mutant p53 alleles (G245S, R248W, R273H, and R273C) and one containing a non-GOF p53 mutant allele (P151S). The four GOF p53 sublines were able to grow under androgen-depleted conditions, whereas the LNCaP parental line, vector-only line, and the non-GOF line were unable to grow. To investigate the mechanism of the AI growth displayed by the GOF p53 mutants, Western blotting or ELISA were used to examine the expression of the androgen receptor (AR), the AR regulated prostate-specific antigen (PSA), as well as Akt and Bcl-2 under androgen-depleted conditions. On androgen ablation, the levels of AR decreased in the four GOF p53 sublines compared with the control lines. This decreased AR expression was accompanied by attenuated receptor activity, because a decrease in prostate-specific antigen levels compared with parental LNCaP cells was also observed. Levels of phosphorylated Akt increased in both the GOF p53 sublines and the control lines. Bcl-2 remains unchanged or showed reduced expression in all of the cell lines in the absence of androgen compared to the presence of androgen. These observations suggest that GOF p53 mutants mediate the AI growth of LNCaP cells in an AR-independent fashion, and that both Akt and Bcl-2 are not involved in this process. PMID- 12727845 TI - PP1 inhibitor induces degradation of RETMEN2A and RETMEN2B oncoproteins through proteosomal targeting. AB - RET tyrosine kinase oncoproteins are potential targets for anticancer therapy. We show here that along with the inhibition of RET tyrosine phosphorylation, the pyrazolo-pyrimidine inhibitor PP1 induces RETMEN2A and RETMEN2B oncoprotein destruction. In fact, as a consequence of PP1 treatment, RET oncoproteins translocate from the outer limiting membrane to inner cellular compartments and are rapidly addressed to the degradative pathway. The cleavage of RET oncoproteins is associated with an impairment of RET mitogenic signaling pathways that causes a reversion of the oncogenic transformation and establishes a long term cytostatic effect. By using specific inhibitors of both the proteosome and the lysosome, we assessed that PP1 targets RET oncoproteins to proteosomal, rather than lysosomal, degradation. In this context of studies, we interestingly demonstrated that RETMEN2A and RETMEN2B receptors are constitutively ubiquitinated and interact with the ubiquitin ligase c-Cbl. Moreover, PP1 does not modify these interactions, although it indeed causes RET dephosphorylation. Therefore, even if the degradative pathway stimulated by the inhibitor appears to be mediated by the proteosome, PP1 does not seem to enhance nor promote receptor ubiquitination. These observations lead us to favor two models for PP1-induced RET oncoprotein degradation: either PP1-mediated RET dephosphorylation per se targets the oncoproteins for destruction or alternatively, PP1 insertion in the RET ATP-binding pocket promotes a mechanism for fast stress-induced degradation. The use of PP1, which therefore acts as a degradation-inducing factor, may represent a promising new strategy to selectively target RET oncogenic products for destruction and holds promise for future medullary thyroid cancer therapy. PMID- 12727846 TI - POU5F1 (OCT3/4) identifies cells with pluripotent potential in human germ cell tumors. AB - Human germ cell tumors (GCTs) may have variable histology and clinical behavior, depending on factors such as sex of the patient, age at clinical diagnosis, and anatomical site of the tumor. Some types of GCT, i.e., the seminomas/germinomas/dysgerminomas and embryonal carcinomas (the stem cell component of nonseminomas), have pluripotent potential, which is demonstrated by their capacity to differentiate into somatic and/or extraembryonic elements. Although embryonal carcinoma cells are intrinsically pluripotent, seminoma/germinoma/dysgerminoma cells, as well as their precursor carcinoma in situ/gonadoblastoma cells, have the phenotype of early germ cells that can be activated to pluripotency. The other types of GCT (teratomas and yolk sac tumors of infants and newborn, dermoid cyst of the ovary, and spermatocytic seminoma of elderly) are composed of (fully) differentiated tissues and lack the appearance of undifferentiated and pluripotent stem cells. OCT3/4, a transcription factor also known as OTF3 and POU5F1, is involved in regulation of pluripotency during normal development and is detectable in embryonic stem and germ cells. We analyzed the presence of POU5F1 in GCT and other tumor types using immunohistochemistry. The protein was consistently detected in carcinoma in situ/gonadoblastoma, seminomas/germinoma/dysgerminoma, and embryonal carcinoma but not in the various types of differentiated nonseminomas. Multitumor tissue microarray analysis covering >100 different tumor categories and 3600 individual cancers verified that POU5F1 expression is specific for particular subtypes of GCT of adults. No protein was observed in GCT of newborn and infants, spermatocytic seminomas, and the various tumors of nongerm cell origin. In addition, no difference in staining pattern was found in chemosensitive and chemoresistant GCT of adults. These results indicate preservation of the link between POU5F1 and pluripotency, as reported during normal development, after malignant transformation. Therefore, POU5F1 immunohistochemistry is an informative diagnostic tool for pluripotent GCT and offers new insights into the histological heterogeneity of this cancer. PMID- 12727847 TI - Differential expression of galectin-3 in pituitary tumors. AB - Galectin-3 (Gal-3), a beta-galactoside-binding protein, has been implicated in a variety of biological functions, including cell proliferation and differentiation, tumor cell adhesion, angiogenesis, apoptosis, tumor progression, and metastasis. We investigated the role of Gal-3 in the development and progression of pituitary tumors. Immunohistochemical and Western blot analysis of normal and neoplastic human pituitaries showed that only lactotroph (PRL) and corticotroph (ACTH) hormone-producing cells and tumors expressed Gal-3. Gal-3 was present in 24 of 38 (63.2%) PRL adenomas, 5 of 6 (83.3%) PRL carcinomas, 19 of 41 (46.3) ACTH adenomas, and 7 of 8 (87.5%) ACTH carcinomas, but not in 112 other pituitary adenomas and carcinomas. Pituitary folliculo-stellate cells, which have macrophage-type functions in the anterior pituitary, also expressed Gal-3. Hyperplastic and neoplastic pituitaries from p27(Kip1) (p27)-null mice, which produce mainly ACTH, showed increased Gal-3 expression levels compared with control mice. Treatment with transforming growth factor beta1, which regulates pituitary cell proliferation, reduced Gal-3 as well as p27 expression levels in cultured HP75 pituitary cells and Gal-3 in cultured pituitary cells from p27-null mice, suggesting that p27 is not necessary for the inhibitory effects of transforming growth factor beta1 on the cell cycle in the pituitary. The role of Gal-3 in pituitary cell function was examined by RNA interference experiments. Inhibition of Gal-3 gene expression by RNA interference decreased HP75 cell proliferation and increased apoptosis. These results indicate that Gal-3 has an important role in pituitary cell proliferation and tumor progression. PMID- 12727849 TI - Loss of p53 in craf-induced transgenic lung adenoma leads to tumor acceleration and phenotypic switch. AB - One of the most frequent malignancies in humans is lung adenocarcinoma.To develop novel diagnostic and therapeutic approaches for the management of this disease, animal models are required. We have used transgenic mice with lung-targeted expression of the CRaf kinase to evaluate genes altered frequently in human lung adenocarcinoma for their effect on tumor progression. Here we report that loss of p53 dramatically accelerates tumor development and induces a phenotypic switch in the target cell from cuboid to a nonciliated columnar morphology. Coexpression of lung epithelial cell markers surfactant protein C and Clara cell antigen suggests that tumor cell dedifferentiation could be involved in this process. The effect of p53 is specific, because loss of one of its target genes, p21(CIP1/WAF1), did not have this effect on cell phenotype although tumor latency was also reduced significantly. Neither loss of p53 nor p21 stimulated acquisition of the metastasis program beyond the stage of bronchiolar extension. This mouse model for pulmonary adenoma and adenocarcinoma should be very helpful for a better understanding of pathogenesis and treatment of this most deadly human cancer. PMID- 12727850 TI - Dipeptidyl peptidase IV overexpression induces up-regulation of E-cadherin and tissue inhibitors of matrix metalloproteinases, resulting in decreased invasive potential in ovarian carcinoma cells. AB - Dipeptidyl peptidase IV (DPPIV/CD26) is a multifunctional cell surface aminopeptidase that is widely expressed in different cell types. Our previous study demonstrated a possible link between DPPIV expression and decreased i.p. dissemination and loss of invasive potential of ovarian carcinoma. In this report, we examined the mechanisms of the anti-invasive ability of DPPIV in greater detail. Expression of E-cadherin and beta-catenin was positively correlated with DPPIV expression among five independent ovarian carcinoma cell lines. The introduction of DPPIV cDNA into an ovarian carcinoma cell line (SKOV3) with low DPPIV expression enhanced the expression of E-cadherin and beta-catenin, with a cellular morphological change from a fibroblastic and motile phenotype to an epithelial phenotype. In addition, matrix metalloproteinase 2 and membrane type 1 matrix metalloproteinase, important markers associated with invasive and metastatic potential, were remarkably reduced. In contrast, tissue inhibitors of matrix metalloproteinases were up-regulated by DPPIV transfection. Furthermore, suppression of the phosphorylation levels of mitogen-activated protein kinase isoform, extracellular signal-regulated kinase, was observed in DPPIV overexpressing cells. To our knowledge, this is the first evidence that increasing DPPIV expression may contribute to prolonged survival by up-regulation of E-cadherin and tissue inhibitors of matrix metalloproteinases. PMID- 12727848 TI - Elevated expression of 12/15-lipoxygenase and cyclooxygenase-2 in a transgenic mouse model of prostate carcinoma. AB - Changes in expression of arachidonic acid (AA) metabolizing enzymes are implicated in the development and progression of human prostate carcinoma (Pca). Transgenic mouse models of Pca that progress from high-grade prostatic intraepithelial neoplasia (HGPIN) to invasive and metastatic carcinoma could facilitate study of the regulation and function of these genes in Pca progression. Herein we characterize the AA-metabolizing enzymes in transgenic mice established with a prostate epithelial-specific long probasin promoter and the SV40 large T antigen (LPB-Tag mice) that develop extensive HGPIN and invasive and metastatic carcinoma with neuroendocrine (NE) differentiation. Murine 8 lipoxygenase (8-LOX), homologue of the 15-LOX-2 enzyme that is expressed in benign human prostatic epithelium and reduced in Pca, was not detected in wild type or LPB-Tag prostates as determined by enzyme assay, reverse transcription PCR, and immunohistochemistry. The most prominent AA metabolite in mouse prostate was 12-HETE. Wild-type prostate (dorsolateral lobe) converted 1.6 +/- 0.5% [(14)C]AA to 12-HETE (n = 7), and this increased to 8.0 +/- 4.4% conversion in LPB-Tag mice with HGPIN (n = 13). Quantitative real-time reverse transcription PCR and immunostaining correlated the increased 12-HETE synthesis with increased neoplastic epithelial expression of 12/15-LOX, the leukocyte-type (L) of 12-LOX and the murine homologue of human 15-LOX-1. Immunostaining showed increased L12 LOX in invasive carcinoma and approximately one-half of metastatic foci. COX-2 mRNA was detectable in neoplastic prostates with HGPIN but not in wild-type prostate. By immunostaining, COX-2 was increased in the neoplastic epithelium of HGPIN but was absent in foci of invasion and metastases. We conclude that (a) AA metabolism in wild-type mouse prostate differs from humans in the basal expression of LOXs (15-LOX-2 in human, absence of its 8-LOX homologue in mouse prostate); (b) increased expression of 12/15-LOX in HGPIN and invasive carcinoma of the LPB-Tag model is similar to the increased 15-LOX-1 in high-grade human Pca; and (c) the LPB-Tag model shows increased COX-2 in HGPIN, and therefore, it may allow additional definition of the role of this enzyme in the subset of human HGPINs or other precursor lesions that are COX-2 positive, as well as investigation of its contribution to neoplastic cell proliferation and tumor angiogenesis in Pca. PMID- 12727851 TI - Inhibition of aggressiveness of metastatic mouse mammary carcinoma cells by the beta2-chimaerin GAP domain. AB - The biological and functional properties of beta2-chimaerin, a novel phorbol ester/diacylglycerol receptor unrelated to protein kinase C isozymes, are largely unknown. It has previously been established that beta2-chimaerin accelerates the hydrolysis rate of GTP from Rac1 in vitro, leading to the inactivation of this GTPase, which plays important roles in the control of actin cytoskeleton organization, proliferation, motility, and invasiveness. To explore the potential role of beta2-chimaerin in invasion and metastasis, we generated stable transfectants for its catalytic domain (the beta-GAP domain) in F3II murine mammary carcinoma cells. Reduced Rac-GTP levels were observed upon stimulation with epidermal growth factor in the beta-GAP clones compared with control cells. Moreover, a marked alteration in actin polymerization in response to epidermal growth factor was observed in the beta-GAP clones, suggesting impairment of Rac dependent responses. The beta-GAP transfectants also evidenced slower growth rates and a striking reduction in their migratory properties. Adenoviral delivery of the beta-GAP domain into F3II cells also led to reduced proliferative and migratory responses. Importantly, significant differences were found between beta GAP transfectants and control cells regarding their tumorigenic and metastatic properties after s.c. inoculation in syngeneic BALB/c mice. Tumors originating from beta-GAP transfectants showed a significantly lower growth rate and reduced invasive ability; in addition, a lower incidence of spontaneous lung metastases was observed. Our results indicate that beta2-chimaerin impairs key steps in the metastatic cascade and provide evidence for a rational modulation of the Rac signaling pathway in cancer treatment. PMID- 12727853 TI - PTEN decreases in vivo vascularization of experimental gliomas in spite of proangiogenic stimuli. AB - Approximately 30-40% of malignant glial tumors exhibit mutations in the tumor suppressor gene, PTEN/MMAC. Additionally, these tumors are associated with (a) mutations in epidermal growth factor receptor (EGFR), leading to a pro-oncogenic constitutive activation, as well as amplification of its gene, and/or (b) mutations in p53, disrupting normal cellular homeostatic processes. Whereas PTEN/MMAC has been shown to possess antiangiogenic action, constitutively active EGFR or p53 gene defects have been associated with proangiogenic action. In this article, we asked if PTEN/MMAC gene transfer into human glioma cells that possess inactivating mutations of the PTEN/MMAC gene but also express either constitutively active EGFR (U87DeltaEGFR cells) or possess an inactivating mutation of p53 (U251 cells) still display inhibited angiogenesis in orthotopic and ectopic models of gliomas. Human glioma xenografts treated with PTEN/MMAC gene transfer exhibited significantly decreased vascularity both in an orthotopic and in an ectopic model. Taken in combination, these results provide strong evidence of PTEN/MMAC's role in regulating glioma angiogenesis even in the presence of strong proangiogenic signals provided by constitutive EGFR activation or p53 inactivation. PMID- 12727852 TI - Membrane type-1-matrix metalloproteinase expressed by prostate carcinoma cells cleaves human laminin-5 beta3 chain and induces cell migration. AB - Degradation of the extracellular matrix by proteolytic enzymes is a central aspect of physiological and pathologic tissue-remodeling processes such as trophoblastic implantation, wound healing, and tumor invasion. We have hypothesized that prostate adenocarcinoma cell invasion through the normal basal lamina is attributable in part to metalloproteinase-induced cleavage of laminin-5 (Ln-5) and enhanced motility of the cancer cells. We studied the role of membrane type-1-matrix metalloproteinase (MT1-MMP) expressed on the surface of prostate tumor cells in cleaving Ln-5 and enhancing the migration of prostate tumor cells. We also determined the nature of the MT1-MMP cleavage of human Ln-5 and how this altered Ln-5 changes the migration of prostate carcinoma cells. We found that human MT1-MMP cleaves purified human Ln-5 to an 80-kDa fragment. Mass spectrometry analyses of the 80-kDa cleaved product by trypsin and chymotrypsin gave 14 and 9 different peptide sequences, respectively, that were identical to the expected amino acid sequence of the Ln-5-beta3 chain. The recovered peptides represent 14.4% (trypsin) and 10.3% (chymotrypsin) of Ln-5-beta3 chain by amino acid count. Both trypsin and chymotrypsin digestion of MT1-MMP-cleaved product of Ln-5 did not show any other peptides that were identical to the other chains of Ln-5. Using a linear migration assay we found that the Ln-5 cleaved by MT1-MMP enhanced the migration of DU-145 prostate carcinoma cells by 2-fold compared with uncleaved Ln-5. The use of blocked antisense MT1-MMP oligonucleotides inhibited the migration of DU-145 cells on Ln-5. We also found that the prostate carcinoma cells expressing high levels of MT1-MMP, such as PC3N and PPC, demonstrated enhanced migration on human Ln-5-coated substrate, and this migration was inhibited using blocked antisense MT1-MMP oligonucleotides. In conclusion, this is a novel and important finding where we have shown that beta3-chain is cleaved by MT1-MMP, and this cleavage enhances migration of prostate cancer cells. PMID- 12727854 TI - Overexpression of vimentin: role in the invasive phenotype in an androgen independent model of prostate cancer. AB - The androgen-sensitive LNCaP prostate cancer cell line is less invasive than hormone-insensitive lines. CL1, an aggressive, hormone-insensitive LNCaP subline derived by continuous passaging in hormone-depleted medium, was compared with the parental cell line by cDNA microarray analysis. The gene coding for the intermediate filament protein vimentin was found to be highly up-regulated in the CL1 subline. This difference was confirmed by Northern and Western blots and visualized by immunofluorescence microscopy. To assess the contribution of vimentin to the invasive phenotype, LNCaP cells were stably transfected to overexpress vimentin, and the CL1 cells were transfected with vimentin antisense construct. The invasiveness of the transfected cells was tested using an in vitro invasion assay. We were able to demonstrate that decreasing vimentin expression in the constitutively vimentin-expressing CL1 cells led to a significant decrease in their invasiveness but that forcing expression of vimentin in the LNCaP cells did not augment their invasiveness. These findings imply that vimentin expression contributes to the invasive phenotype but cannot confer it alone. PMID- 12727855 TI - Hypermethylation of the Pendred syndrome gene SLC26A4 is an early event in thyroid tumorigenesis. AB - Expression of the recently cloned Pendred syndrome gene SLC26A4 or PDS has been found to be decreased or even absent in various thyroid tumors. To explore the underlying mechanism, we conducted DNA sequencing and methylation-specific PCR studies in 64 primary thyroid tumors and 6 thyroid cell lines. We found aberrant hypermethylation of the SLC26A4 gene in 44% of histologically benign adenomas, 46% of follicular thyroid cancers, 71% of papillary thyroid cancers, 71% of anaplastic thyroid cancers, and 100% of cell lines. A reciprocal relationship between methylation and expression of the gene was confirmed in cell lines and thyroid tissues. We have thus demonstrated epigenetic changes as a new mechanism in altering the SLC26A4 gene function, in addition to genetic mutation in Pendred syndrome. SLC26A4 gene methylation in benign adenomas and the relatively well differentiated WRO cell line suggest that this alteration is an early event in thyroid tumorigenesis. PMID- 12727856 TI - Methylation of the thyroid-stimulating hormone receptor gene in epithelial thyroid tumors: a marker of malignancy and a cause of gene silencing. AB - Thyroid-stimulating hormone receptor (TSHR) expression is frequently silenced in epithelial thyroid cancers associated with decreased or absent TSH-promoted iodine uptake. To study the underlying molecular mechanism of decreased TSHR expression, we examined the methylation status of the TSHR gene promoter by sequencing bisulfite-treated DNA from thyroid tumors. After identification of methylated sites by sequencing bisulfite-treated DNA, we used methylation specific polymerase chain reaction and found frequent CpG methylation in papillary thyroid cancer (23 of 39 patients; 59%) and follicular thyroid cancers (7 of 15 patients; 47%). In contrast, we saw no methylation in normal thyroid tissues and benign adenomas (0 of 8 patients; 0%). In human thyroid tumor cell lines, we observed that TSHR was normally expressed at the protein and mRNA level in cells where the TSHR gene was unmethylated, whereas it was silenced in cell lines where the TSHR promoter was hypermethylated. Treatment of the latter cells with a demethylating agent partially restored TSHR expression. We thus demonstrate aberrant methylation of human TSHR as a likely molecular pathway responsible for the silencing of this gene in thyroid cancers. We propose that methylation of TSHR may provide a novel diagnostic marker of malignancy and a basis for potential use of demethylating agents in conjunction with TSH-promoted radioiodine therapy for epithelial thyroid cancers. PMID- 12727857 TI - Tumor angiogenesis modulates leukocyte-vessel wall interactions in vivo by reducing endothelial adhesion molecule expression. AB - The expression of endothelial cell (EC) adhesion molecules involved in leukocyte vessel wall interactions is suppressed in malignancies. In the present study, we investigated in vivo the regulation of leukocyte-vessel wall interactions by the presence of a tumor. By means of intravital microscopy, tumor necrosis factor alpha-stimulated leukocyte-vessel wall interactions were studied in ear skin microvessels of nude mice bearing small human LS174T colon carcinomas and in C57Bl/6 mice bearing murine B16F10 melanomas. Leukocyte-vessel wall interactions were studied both within and outside small tumors growing in the ear, and in ear microvessels of mice with a large tumor growing on their flank. Tumor-free mice were used as controls. Compared with values measured at the edge of the ear and in the contralateral ear, leukocyte adhesion was found to be diminished significantly in vessels inside the ear tumor in both mouse models. This reduction disappeared with increasing distance from the tumor. Surprisingly, the level of leukocyte adhesion in ear venules of mice with a large flank tumor was also reduced significantly. Leukocyte rolling, i.e., the step preceding adhesion, was not influenced by the presence of a tumor in nude mice, but was down regulated in immune-competent C57Bl/6 mice. Treatment of mice bearing a small ear tumor with a humanized antivascular endothelial growth factor antibody prevented the down-regulation of leukocyte-vessel wall interactions inside the tumor vessels compared with the nontreated group. Fluorescence-activated cell sorter analysis showed that isolated tumor ECs have suppressed levels of intercellular adhesion molecule 1 as compared with ECs from normal mouse tissues. In cultured b.END5 cells the tumor necrosis factor alpha-induced up-regulation of intercellular adhesion molecule 1 and vascular cell adhesion molecule 1 was reduced in ECs that were preincubated with basic fibroblast growth factor or vascular endothelial growth factor. The current results may have an impact on the effectiveness of clinical immunotherapeutic treatment protocols, because immune effector cells may not be able to enter tumor tissue. PMID- 12727858 TI - Vascular endothelial growth factor gene expression in colon cancer cells exposed to prostaglandin E2 is mediated by hypoxia-inducible factor 1. AB - Prostaglandin E(2) (PGE(2)) has been implicated as an inducer of angiogenesis in human colon cancer. Here, we demonstrate that PGE(2) exposure induces the expression of vascular endothelial growth factor (VEGF) mRNA in HCT116 human colon carcinoma cells that is mediated by the transcriptional activator hypoxia inducible factor 1 (HIF-1). PGE(2) exposure induces the phosphorylation of extracellular signal-regulated kinase (ERK) and AKT. Pharmacologic inhibition of ERK phosphorylation blocks the induction of VEGF mRNA and HIF-1alpha protein expression in response to PGE(2) stimulation. Inhibition of C-SRC tyrosine kinase activity also blocks PGE(2)-induced HIF-1alpha protein and VEGF mRNA expression without blocking ERK phosphorylation. In contrast, phosphorylation of AKT is dependent on ERK and C-SRC activity. Thus, the activity of multiple signal transduction pathways is required for the HIF-1-mediated induction of VEGF expression in colon cancer cells exposed to PGE(2). PMID- 12727859 TI - CrkI adapter protein modulates cell migration and invasion in glioblastoma. AB - The human crk gene is translated into crkI and crkII by alternative splicing. crkII mRNA was detected both in normal brain and glioblastoma tissues, whereas crkI mRNA levels were quite low in normal brain and up-regulated in glioblastoma tissues. Expression of CrkI but not CrkII in glioblastoma U87MG cells induced transformation that stimulated cell migration and invasion concomitant with tyrosine phosphorylation of p130 Crk-associated substrate. N-cadherin-mediated signal transduction, which was essential for invasion by U87MG cells, was no longer required for CrkI-transformed cells. These results suggest that CrkI contributes to malignancy of glioblastoma by inducing phosphorylation of p130 Crk associated substrate. PMID- 12727860 TI - Reactivity with A monoclonal antibody to Epstein-Barr virus (EBV) nuclear antigen 1 defines a subset of aggressive breast cancers in the absence of the EBV genome. AB - Previous studies have suggested that common breast cancers are associated with EBV. We used a highly sensitive quantitative real-time PCR method to screen whole tumor sections of breast cancers for the presence of the EBV genome. EBV DNA was detected in 19 of 92 (21%) tumors, but viral load was very low in positive samples (mean = 1.1 copy EBV/1000 cells, maximum = 7.1 copies EBV/1000 cells). Importantly, quantitative real-time PCR failed to detect the EBV genome in microdissected tumor cells from any case. Using a monoclonal antibody (2B4-1) reactive against the EBV nuclear antigen-1, we noted strong staining of tumor nuclei in a proportion of those breast cancers that had tested negative for the presence of the EBV genome. Because nuclear staining with the 2B4-1 antibody was previously observed more frequently in poor prognosis breast cancers, we examined a larger series of breast cancers with complete clinical follow-up. Strong punctate staining of tumor cell nuclei was observed in 47 of 153 (31%) breast cancers; 2B4-1-positive tumors were significantly more likely to be ER-negative (P < 0.0001), to be of higher grade (P = 0.001) and larger (P = 0.03), to involve more regional lymph nodes (P = 0.01), and to have higher Nottingham Prognostic Index scores (P = 0.0003). Conclusions are: (a) EBV can be regularly detected in whole sections of breast cancers but viral copy number is very low; (b) in these cases, tumor cells do not harbor virus; and (c) reactivity with the monoclonal antibody 2B4-1 is detectable in the absence of the EBV genome and is strongly associated with ER-negative breast tumors and with prognostically unfavorable disease. Additional studies should be directed to the identification of this protein and to elucidation of its role in breast cancer. PMID- 12727861 TI - Crawling into a new era-the Dictyostelium genome project. AB - The social amoeba Dictyostelium discoideum is a well-established model organism for the study of basic aspects of differentiation, signal transduction, phagocytosis, cytokinesis and cell motility. Its genome is being sequenced by an international consortium using a whole chromosome shotgun approach. The pacemaker of the D.discoideum genome project has been chromosome 2, the largest chromosome, which at 8 Mb represents approximately 25% of the genome and whose sequence and analysis have been published recently. Chromosomes 1 and 6 are close to being finished. To accelerate completion of the genome sequence, the next step in the project will be a whole-genome assembly followed by the analysis of the complete gene content. The completed genome sequence and its analysis provide the basis for genome-wide functional studies. It will position Dictyostelium at the same level as other model organisms and further enhance its experimental attractiveness. PMID- 12727862 TI - In and out of Torso RTK signalling. PMID- 12727863 TI - Functional aspects of protein mono-ADP-ribosylation. AB - Mono-ADP-ribosylation is the enzymatic transfer of ADP-ribose from NAD(+) to acceptor proteins. It is catalysed by cellular ADP-ribosyltransferases and certain bacterial toxins. There are two subclasses of cellular enzymes: the ectoenzymes that modify targets such as integrins, defensin and other cell surface molecules; and the intracellular enzymes that act on proteins involved in cell signalling and metabolism, such as the beta-subunit of heterotrimeric G proteins, GRP78/BiP and elongation factor 2. The genes that encode the ectoenzymes have been cloned and their protein products are well characterized, yet little is known about the intracellular ADP-ribosyltransferases, which may be part of a novel protein family with an important role in regulating cell function. ADP-ribosylation usually leads to protein inactivation, providing a mechanism to inhibit protein functions in both physiological and pathological conditions. PMID- 12727864 TI - Ferritins, iron uptake and storage from the bacterioferritin viewpoint. AB - Ferritins constitute a broad superfamily of iron storage proteins, widespread in all domains of life, in aerobic or anaerobic organisms. Ferritins isolated from bacteria may be haem-free or contain a haem. In the latter case they are called bacterioferritins. The primary function of ferritins inside cells is to store iron in the ferric form. A secondary function may be detoxification of iron or protection against O(2) and its radical products. Indeed, for bacterioferritins this is likely to be their primary function. Ferritins and bacteroferritins have essentially the same architecture, assembling in a 24mer cluster to form a hollow, roughly spherical construction. In this review, special emphasis is given to the structure of the ferroxidase centres with native iron-containing sites, since oxidation of ferrous iron by molecular oxygen takes place in these sites. Although present in other ferritins, a specific entry route for iron, coupled with the ferroxidase reaction, has been proposed and described in some structural studies. Electrostatic calculations on a few selected proteins indicate further ion channels assumed to be an entry route in the later mineralization processes of core formation. PMID- 12727865 TI - VirB11 ATPases are dynamic hexameric assemblies: new insights into bacterial type IV secretion. AB - The coupling of ATP binding/hydrolysis to macromolecular secretion systems is crucial to the pathogenicity of Gram-negative bacteria. We reported previously the structure of the ADP-bound form of the hexameric traffic VirB11 ATPase of the Helicobacter pylori type IV secretion system (named HP0525), and proposed that it functions as a gating molecule at the inner membrane, cycling through closed and open forms regulated by ATP binding/hydrolysis. Here, we combine crystal structures with analytical ultracentrifugation experiments to show that VirB11 ATPases indeed function as dynamic hexameric assemblies. In the absence of nucleotide, the N-terminal domains exhibit a collection of rigid-body conformations. Nucleotide binding 'locks' the hexamer into a symmetric and compact structure. We propose that VirB11s use the mechanical leverage generated by such nucleotide-dependent conformational changes to facilitate the export of substrates or the assembly of the type IV secretion apparatus. Biochemical characterization of mutant forms of HP0525 coupled with electron microscopy and in vivo assays support such hypothesis, and establish the relevance of VirB11s ATPases as drug targets against pathogenic bacteria. PMID- 12727866 TI - CFTR directly mediates nucleotide-regulated glutathione flux. AB - Studies have shown that expression of cystic fibrosis transmembrane conductance regulator (CFTR) is associated with enhanced glutathione (GSH) efflux from airway epithelial cells, implicating a role for CFTR in the control of oxidative stress in the airways. To define the mechanism underlying CFTR-associated GSH flux, we studied wild-type and mutant CFTR proteins expressed in Sf9 membranes, as well as purified and reconstituted CFTR. We show that CFTR-expressing membrane vesicles mediate nucleotide-activated GSH flux, which is disrupted in the R347D pore mutant, and in the Walker A K464A and K1250A mutants. Further, we reveal that purified CFTR protein alone directly mediates nucleotide-dependent GSH flux. Interestingly, although ATP supports GSH flux through CFTR, this activity is enhanced in the presence of the non-hydrolyzable ATP analog AMP-PNP. These findings corroborate previous suggestions that CFTR pore properties can vary with the nature of the nucleotide interaction. In conclusion, our data demonstrate that GSH flux is an intrinsic function of CFTR and prompt future examination of the role of this function in airway biology in health and disease. PMID- 12727867 TI - An H-bond between two residues from different loops of the acetylcholine binding site contributes to the activation mechanism of nicotinic receptors. AB - The molecular mechanisms of nicotinic receptor activation are still largely unknown. The crystallographic structure of the acetylcholine binding protein (AChBP) reveals a single H-bond between two different acetylcholine binding loops. Within these homologous loops we systematically introduced alpha4 residues into the alpha7/5HT(3) chimeric receptor and found that the single point mutations G152K (loop B) and P193I (loop C) displayed a non-additive increase of equilibrium binding affinity for several agonists compared with the double mutant G152K/P193I. In whole-cell patch-clamp recordings, G152K, P193I and G152K/P193I mutants displayed an increase up to 5-fold in acetylcholine potency with a large decrease of the apparent Hill coefficients (significantly smaller than one). Concomitantly, the G152K/P193I mutant showed a dramatic loss of high-affinity alpha-bungarotoxin binding (100-fold decrease), thus pinpointing a new contact area for the toxin. Fitting the data with an allosteric-kinetic model, together with molecular dynamic simulations, suggests that the presence of the inter backbone H-bond between positions 152 and 193, revealed in alpha4 and in alpha7 double mutant but not in alpha7, coincides with a large stabilization of both open and desensitized states of nicotinic receptors. PMID- 12727868 TI - Functional analysis of AtHKT1 in Arabidopsis shows that Na(+) recirculation by the phloem is crucial for salt tolerance. AB - Two allelic recessive mutations of Arabidopsis, sas2-1 and sas2-2, were identified as inducing sodium overaccumulation in shoots. The sas2 locus was found (by positional cloning) to correspond to the AtHKT1 gene. Expression in Xenopus oocytes revealed that the sas2-1 mutation did not affect the ionic selectivity of the transporter but strongly reduced the macro scopic (whole oocyte current) transport activity. In Arabidopsis, expression of AtHKT1 was shown to be restricted to the phloem tissues in all organs. The sas2-1 mutation strongly decreased Na(+) concentration in the phloem sap. It led to Na(+) overaccumulation in every aerial organ (except the stem), but to Na(+) underaccumulation in roots. The sas2 plants displayed increased sensitivity to NaCl, with reduced growth and even death under moderate salinity. The whole set of data indicates that AtHKT1 is involved in Na(+) recirculation from shoots to roots, probably by mediating Na(+) loading into the phloem sap in shoots and unloading in roots, this recirculation removing large amounts of Na(+) from the shoot and playing a crucial role in plant tolerance to salt. PMID- 12727869 TI - A novel NADH kinase is the mitochondrial source of NADPH in Saccharomyces cerevisiae. AB - Mitochondria require NADPH for anti-oxidant protection and for specific biosynthetic pathways. However, the sources of mitochondrial NADPH and the mechanisms of maintaining mitochondrial redox balance are not well understood. We show here that in Saccharomyces cerevisiae, mitochondrial NADPH is largely provided by the product of the POS5 gene. We identified POS5 in a S.cerevisiae genetic screen for hyperoxia-sensitive mutants, or cells that cannot survive in 100% oxygen. POS5 encodes a protein that is homologous to NAD(+) and NADH kinases, and we show here that recombinant Pos5p has NADH kinase activity. Pos5p is localized to the mitochondrial matrix of yeast and appears to be important for several NADPH-requiring processes in the mitochondria, including resistance to a broad range of oxidative stress conditions, arginine biosynthesis and mitochondrial iron homeostasis. Pos5p represents the first member of the NAD(H) kinase family that has been identified as an important anti-oxidant factor and key source of the cellular reductant NADPH. PMID- 12727870 TI - A conserved ER targeting motif in three families of lipid binding proteins and in Opi1p binds VAP. AB - Intracellular lipid traffic is mediated both by membrane vesicles and by a number of non-vesicular pathways facilitated by cytoplasmic lipid binding proteins. For these proteins to act effectively they must be targeted accurately to specific membranes. Here we identify a novel short conserved determinant called the FFAT motif that is shared by several seemingly unrelated lipid binding proteins and is also found in Opi1p, a transcriptional regulator of phospholipid synthesis in yeast. FFAT motifs act as membrane- targeting determinants by their direct interaction with homologues of VAMP-associated protein (VAP), a conserved endoplasmic reticulum (ER) protein. In budding yeast, all four proteins with FFAT motifs interact with Scs2p, a homologue of VAP, to target the ER to some extent. The precise intracellular distribution of each of these proteins depends on the integration of the FFAT-Scs2p interaction with other targeting determinants, and the interaction is functionally significant. We conclude that binding to a VAP homologue is a common mechanism by which proteins with FFAT motifs, most of which are involved in lipid metabolism, target ER membranes. PMID- 12727871 TI - Ras promotes p21(Waf1/Cip1) protein stability via a cyclin D1-imposed block in proteasome-mediated degradation. AB - Ras promotes the accumulation of the cyclin-dependent kinase inhibitor p21(Waf1/Cip1) (p21). Previous studies reported that acute Raf/MEK/ERK activation elevates p21 protein levels by increased transcription. However, we have found that p21 induction in Ras-transformed murine fibroblasts occurs principally by a post-translational mechanism. Chronic activation of the Raf/MEK/ERK pathway blocked proteasome-mediated p21 degradation, resulting in accumulation of p21 protein with an elevated half-life. The stabilization of p21 by Ras was accompanied by high levels of p21-associated cyclin D1 and, similarly to Ras, cyclin D1 was sufficient to inhibit the proteasome-mediated p21 degradation. Knock-down of cyclin D1 by RNA interference confirmed that Ras-induced p21 stabilization was dependent upon cyclin D1 expression. We show that p21 directly binds to the C8alpha subunit of the 20S proteasome complex and that by competing for binding, cyclin D1 inhibits p21 degradation by purified 20S complexes in vitro. Therefore, we propose that Ras stabilizes p21 by promoting the formation of p21-cyclin D1 complexes that prevent p21 association with, and subsequent degradation by, the 20S proteasome. PMID- 12727872 TI - Sumoylation is involved in beta-catenin-dependent activation of Tcf-4. AB - Sumoylation is involved in mediating protein-protein interactions, subcellular compartmentalization and protein stability. Our analysis of various Wnt signaling molecules revealed that one of them, Tcf-4, is sumoylated at the endogenous level. At least one sumoylation site, Lys297, of Tcf-4 was identified. The sumoylation of Tcf-4 was enhanced by PIASy, a SUMO E3 enzyme, and inhibited by Axam, a desumoylation enzyme. Although PIASy did not affect the interaction of Tcf-4 with beta-catenin or DNA, Tcf-4, SUMO-1 and PIASy were co-localized in the nucleus and present in a complex in the PML body. PIASy enhanced beta-catenin dependent transcriptional activity of Tcf-4, whereas Axam inhibited it. Reduction of the protein level of Axam by RNA interference led to an increase in sumoylation of Tcf-4 and activation of Tcf-4. Furthermore, beta-catenin and PIASy activated Tcf-4(K297R), in which Lys297 was changed to arginine, less than wild type Tcf-4. These results suggest that sumoylation of Tcf-4 is involved in beta catenin-dependent and Tcf-4-mediated gene expression in the Wnt signaling pathway. PMID- 12727873 TI - Importin alpha-regulated nucleation of microtubules by TPX2. AB - The importin alpha-regulated microtubule-associated protein TPX2 is known to be critical for meiotic and mitotic spindle formation in vertebrates, but its detailed mechanism of action and regulation is not understood. Here, the site of interaction on TPX2 for importin alpha is mapped. A TPX2 mutant that cannot bind importin alpha is constitutively active in the induction of microtubule containing aster-like structures in Xenopus egg extract, demonstrating that no other importin alpha or RanGTPase target is required to mediate microtubule assembly in this system. Further, recombinant TPX2 is shown to induce the formation and bundling of microtubules in dilute solutions of pure tubulin. In this purified system, importin alpha prevents TPX2-induced microtubule formation, but not TPX2-tubulin interaction or microtubule bundling. This demonstrates that TPX2 has more than one mode of interaction with tubulin and that only one of these types of interaction is abolished by importin alpha. The data suggest that the critical early function in spindle formation regulated by importin alpha is TPX2-mediated microtubule nucleation. PMID- 12727874 TI - Domain organization and structure-function relationship of the HET-s prion protein of Podospora anserina. AB - The [Het-s] infectious element of the fungus Podospora anserina is a prion protein involved in a genetically controlled cell death reaction termed heterokaryon incompatibility. Previous analyses indicate that [Het-s] propagates as a self-perpetuating amyloid aggregate. The HET-s protein is 289 amino acids in length. Herein, we identify the region of the HET-s protein that is responsible for amyloid formation and prion propagation. The region of HET-s spanning residues 218-289 forms amyloid fibers in vitro and allows prion propagation in vivo. Conversely, a C-terminal deletion in HET-s prevents amyloid aggregation in vitro and prion propagation in vivo, and abolishes the incompatibility function. In the soluble form of HET-s, the region from residue 1 to 227 forms a well folded domain while the C-terminal region is highly flexible. Together, our data establish a domain structure-function relationship for HET-s amyloid formation, prion propagation and incompatibility activity. PMID- 12727875 TI - Rapid invasion of host cells by Toxoplasma requires secretion of the MIC2-M2AP adhesive protein complex. AB - Vertebrate cells are highly susceptible to infection by obligate intracellular parasites such as Toxoplasma gondii, yet the mechanism by which these microbes breach the confines of their target cell is poorly understood. While it is thought that Toxoplasma actively invades by secreting adhesive proteins from internal organelles called micronemes, no genetic evidence is available to support this contention. Here, we report successful disruption of M2AP, a microneme protein tightly associated with an adhesive protein called MIC2. M2AP knockout parasites were >80% impaired in host cell entry. This invasion defect was likely due to defective expression of MIC2, which partially accumulated in the parasite endoplasmic reticulum and Golgi. M2AP knockout parasites were also unable to rapidly secrete MIC2, an event that normally accompanies parasite attachment to a target cell. These findings indicate a critical role for the MIC2 M2AP protein complex in parasite invasion. PMID- 12727876 TI - Cdc2-mediated phosphorylation of Kid controls its distribution to spindle and chromosomes. AB - The chromokinesin Kid is important in chromosome alignment at the metaphase plate. Here, we report that Kid function is regulated by phosphorylation. We identify Ser427 and Thr463 as M phase-specific phosphorylation sites and Cdc2 cyclin B as a Thr463 kinase. Kid with a Thr463 to alanine mutation fails to be localized on chromosomes and is only detected along spindles, although it retains the ability to bind DNA or chromosomes. Localization of rigor-type mutant Kid, which shows nucleotide-independent microtubule association, is also confined to the spindle, implying that strong association of Kid with the spindle can sequester it from chromosomes. T463A substitution in DNA-binding domain-truncated Kid consistently enhances its spindle localization. At physiological ionic strength, unphosphorylated Kid shows ATP-independent microtubule association, whereas Thr463-phosphorylated Kid shows ATP dependency. Moreover, the stalk region of unphosphorylated Kid interacts with microtubules and the interaction is weakened when Thr463 is phosphorylated. Our data suggest that phosphorylation on Thr463 of Kid downregulates its affinity for microtubules to ensure reversible association with spindles, allowing Kid to bind chromosomes and exhibit its function. PMID- 12727877 TI - Pro-apoptotic function of HBV X protein is mediated by interaction with c-FLIP and enhancement of death-inducing signal. AB - Despite its implication in the progression of hepatitis B virus (HBV)-associated liver disease, the pro-apoptotic function of HBx protein remains poorly understood. We show that the expression of HBx leads to hyperactivation of caspase-8 and caspase-3 upon treatment with tumor necrosis factor-alpha (TNF alpha) or anti-Fas antibody, and this activation is correlated with the sensitivity to apoptosis. We demonstrate cytoplasmic co-localization and direct interaction between HBx and the cellular FLICE inhibitory protein (c-FLIP), a key regulator of the death-inducing signaling complex (DISC). Deletion analysis shows that the death effector domain 1 (DED1) of c-FLIP is important for the observed interaction. Overexpression of c-FLIP rescued the cells from HBx-mediated apoptosis, with both the full-length HBV genome and HBx expression vectors. Moreover, c-FLIP and caspase-8 inhibitor considerably protected cells from HBx mediated apoptosis. These data suggest that HBx abrogates the apoptosis inhibitory function of c-FLIP and renders the cell hypersensitive towards the TNF alpha apoptotic signal even below threshold concentration. This provides a novel mechanism for deregulation of hepatic cell growth in HBV patients and a new target for intervention in HBV-associated liver cancer and disease. PMID- 12727878 TI - Cyanobacterial circadian clockwork: roles of KaiA, KaiB and the kaiBC promoter in regulating KaiC. AB - Using model strains in which we ectopically express the cyanobacterial clock protein KaiC in cells from which the clock genes kaiA, kaiB and/or kaiC are deleted, we found that some features of circadian clocks in eukaryotic organisms are conserved in the clocks of prokaryotic cyanobacteria, but others are not. One unexpected difference is that the circadian autoregulatory feedback loop in cyanobacteria does not require specific clock gene promoters as it does in eukaryotes, because a heterologous promoter can functionally replace the kaiBC promoter. On the other hand, a similarity between eukaryotic clock proteins and the cyanobacterial KaiC protein is that KaiC is phosphorylated in vivo. The other essential clock proteins KaiA and KaiB modulate the status of KaiC phosphorylation; KaiA inhibits KaiC dephosphorylation and KaiB antagonizes this action of KaiA. Based upon an analysis of clock mutants, we conclude that the circadian period in cyanobacteria is determined by the phosphorylation status of KaiC and also by the degradation rate of KaiC. These observations are integrated into a model proposing rhythmic changes in chromosomal status. PMID- 12727879 TI - KaiB functions as an attenuator of KaiC phosphorylation in the cyanobacterial circadian clock system. AB - In the cyanobacterium Synechococcus elongatus PCC 7942, the KaiA, KaiB and KaiC proteins are essential for generation of circadian rhythms. We quantitatively analyzed the intracellular dynamics of these proteins and found a circadian rhythm in the membrane/cytosolic localization of KaiB, such that KaiB interacts with a KaiA-KaiC complex during the late subjective night. KaiB-KaiC binding is accompanied by a dramatic reduction in KaiC phosphorylation and followed by dissociation of the clock protein complex(es). KaiB attenuated KaiA-enhanced phosphorylation both in vitro and in vivo. Based on these results, we propose a novel role for KaiB in a regulatory link among subcellular localization, protein protein interactions and post-translational modification of Kai proteins in the cyanobacterial clock system. PMID- 12727880 TI - Stimulation of preadipocyte differentiation by steroid through targeting of an HDAC1 complex. AB - Glucocorticoids potentiate the early steps of preadipocyte differentiation and promote obesity in Cushing's syndrome and during prolonged steroid therapy. We show that glucocorticoids stimulate 3T3 L1 preadipocyte differentiation through a non-transcriptional mechanism mediated through the ligand-binding domain of the glucocorticoid receptor. This enhanced the onset of CCAAT/enhancer binding protein (C/EBPalpha) expression by potentiating its initial transcriptional activation by C/EBPbeta. In the absence of steroid, C/EBPbeta associated with a transcriptional corepressor complex containing mSin3A and histone deacetylase 1 (HDAC1), but lacking HDAC2 and RbAp46/48. HDAC1/mSin3A were recruited to the C/EBPalpha promoter with C/EBPbeta and promoted the deacetylation of histone H4. Steroid induced the specific depletion of this corepressor by targeting the HDAC1 within the complex for degradation through the 26S proteasome. Treatment with histone deacetylase inhibitors replaced the effects of steroid treatment on preadipocyte differentiation and C/EBPalpha expression, while overexpression of HDAC1 abrogated the stimulatory effects of steroid. Recapitulation of the glucocorticoid effect by progestin treatment in the presence of the progesterone receptor ligand-binding domain suggests a conserved mechanism relevant to many aspects of steroid-mediated differentiation. PMID- 12727881 TI - A role for cofactor-cofactor and cofactor-histone interactions in targeting p300, SWI/SNF and Mediator for transcription. AB - Transcriptional activation from chromatin by nuclear receptors (NRs) requires multiple cofactors including CBP/p300, SWI/SNF and Mediator. How NRs recruit these multiple cofactors is not clear. Here we show that activation by androgen receptor and thyroid hormone receptor is associated with the promoter targeting of SRC family members, p300, SWI/SNF and the Mediator complex. We show that recruitment of SWI/SNF leads to chromatin remodeling with altered DNA topology, and that both SWI/SNF and p300 histone acetylase activity are required for hormone-dependent activation. Importantly, we show that both the SWI/SNF and Mediator complexes can be targeted to chromatin by p300, which itself is recruited through interaction with SRC coactivators. Furthermore, histone acetylation by CBP/p300 facilitates the recruitment of SWI/SNF and Mediator. Thus, our data indicate that multiple cofactors required for activation are not all recruited through their direct interactions with NRs and underscore a role of cofactor-cofactor interaction and histone modification in coordinating the recruitment of multiple cofactors. PMID- 12727882 TI - Recruitment of human cyclin T1 to nuclear bodies through direct interaction with the PML protein. AB - Human cyclin T1, the cyclin partner of Cdk9 kinase in the positive transcription elongation factor b (P-TEFb), is an essential cellular cofactor that is recruited by the human immunodeficiency virus type 1 (HIV-1) Tat transactivator to promote transcriptional elongation from the HIV-1 long terminal repeat (LTR). Here we exploit fluorescence resonance energy transfer (FRET) to demonstrate that cyclin T1 physically interacts in vivo with the promyelocytic leukaemia (PML) protein within specific subnuclear compartments that are coincident with PML nuclear bodies. Deletion mutants at the C-terminal region of cyclin T1 are negative for FRET with PML and fail to localize to nuclear bodies. Cyclin T1 and PML are also found associated outside of nuclear bodies, and both proteins are present at the chromatinized HIV-1 LTR promoter upon Tat transactivation. Taken together these results suggest that PML proteins regulate Tat- mediated transcriptional activation by modulating the availability of cyclin T1 and other essential cofactors to the transcription machinery. PMID- 12727883 TI - Independent functions of yeast Pcf11p in pre-mRNA 3' end processing and in transcription termination. AB - Pcf11p, an essential subunit of the yeast cleavage factor IA, is required for pre mRNA 3' end processing, binds to the C-terminal domain (CTD) of the largest subunit of RNA polymerase II (RNAP II) and is involved in transcription termination. We show that the conserved CTD interaction domain (CID) of Pcf11p is essential for cell viability. Interestingly, the CTD binding and 3' end processing activities of Pcf11p can be functionally uncoupled from each other and provided by distinct Pcf11p fragments in trans. Impaired CTD binding did not affect the 3' end processing activity of Pcf11p and a deficiency of Pcf11p in 3' end processing did not prevent CTD binding. Transcriptional run-on analysis with the CYC1 gene revealed that loss of cleavage activity did not correlate with a defect in transcription termination, whereas loss of CTD binding did. We conclude that Pcf11p is a bifunctional protein and that transcript cleavage is not an obligatory step prior to RNAP II termination. PMID- 12727884 TI - Regions of GAL4 critical for binding to a promoter in vivo revealed by a visual DNA-binding analysis. AB - Binding of transcriptional activators to specific sites on DNA, mediated by their DNA-binding domain, is a key regulatory point in transcriptional regulation. With a GFP-based microscopic assay, we investigated how the prototypical activator GAL4 effectively binds to a promoter in living yeast cells. We show that GAL4 relies on a previously unrevealed mechanism involving 'DNA-binding enhancers' (DBEs), the regions of GAL4 that assist DNA-binding domain association with DNA. GAL4 contains two DBEs, one, but not the other, physically overlapping the principal transcriptional activation domain. Either of the DBEs, however, can function independently of transcriptional activation, indicating a discrete mechanism responsible for DNA-binding enhancement. The effect of DBEs, while not limited to natural target promoters, is still not universal and can be profoundly affected by the binding-site context. The GAL4 DBEs can also enhance promoter binding of an unrelated DNA-binding domain, and possibly represent a new modular functional unit responsible for effective binding of diverse regulatory factors to DNA in vivo. PMID- 12727885 TI - OBF1 enhances transcriptional potential of Oct1. AB - The POU transcription factors Oct1 and Oct2 bind to DNA in various monomer and dimer configurations. Depending on the DNA sequence to which they bind, the dimers are arranged in configurations that are either accessible (PORE sequence) or inaccessible (MORE sequence) to the B-cell-specific cofactor OBF1 (OcaB, Bob1). As shown previously, the MORE and related sequences (such as the heptamer/octamer motif) are found in immunoglobulin heavy chain promoters. Here we show that the expression of Osteopontin, which contains a PORE sequence in its enhancer region, depends on the presence of OBF1 in B cells. OBF1 alleviates DNA sequence requirements of the Oct1 dimer on PORE-related sequences in vitro. Furthermore, OBF1 stabilizes POU dimer-DNA interactions and overrides Oct1 interface mutations, which abolish PORE-mediated dimerization without OBF1. Our data indicate that the PORE-type Oct1 or Oct2 dimer, rather than the monomer, is the primary target of the cofactor OBF1. Based on our biochemical data, we propose a mode of OBF1-Oct1 dimer interaction, suggesting a novel arrangement of the subdomain connectivities. PMID- 12727886 TI - An unusual MAP kinase is required for efficient penetration of the plant surface by Ustilago maydis. AB - In Ustilago maydis, pathogenic development is controlled by a heterodimer of the two homeodomain proteins bW and bE. We have identified by RNA fingerprinting a b regulated gene, kpp6, which encodes an unusual MAP kinase. Kpp6 is similar to a number of other fungal MAP kinases involved in mating and pathogenicity, but contains an additional N-terminal domain unrelated to other proteins. Transcription of the kpp6 gene yields two transcripts differing in length, but encoding proteins of identical mass. One transcript is upregulated by the bW/bE heterodimer, while the other is induced after pheromone stimulation. kpp6 deletion mutants are attenuated in pathogenicity. kpp6(T355A,Y357F) mutants carrying a non-activatable allele of kpp6 are more severely compromised in pathogenesis. These strains can still form appressoria, but are defective in the subsequent penetration of the plant cuticle. Kpp6 is expressed during all stages of the sexual life cycle except mature spores. We speculate that Kpp6 may respond to a plant signal and regulate the genes necessary for efficient penetration of plant tissue. PMID- 12727887 TI - A complex network of regulatory elements in Ikaros and their activity during hemo lymphopoiesis. AB - Regulated expression of Ikaros is critical for normal hemopoiesis and lymphocyte development. To elucidate the mechanisms underlying transcription of Ikaros, tissue-specific DNase I-hypersensitive sites (DHS) were mapped throughout the Ikaros locus, and several promoters were identified. The activity of these regulatory regions was elucidated using an enhanced green fluorescent protein (EGFP) reporter in transgenic mice. Two genomic fragments, each containing a distinct promoter and its associated DHS cluster, were found to be active in the myeloid (DHS-C2 and DHS-C3) and B-cell (DHS-C3) lineages. Although neither of these regulatory regions was active within the majority of differentiating thymocytes and mature T cells, the DHS-C3 region was active at the earliest stages (DN1-DN3) of T-cell differentiation. However, when the DHS-C3 region was combined with the downstream intronic DHS-C6 cluster, its activity was maintained and raised to higher levels at subsequent stages of T-cell differentiation. This combination of regulatory elements provided reporter expression that closely resembles that of endogenous Ikaros during hemo-lymphopoiesis, and it decreased (but did not alleviate) position effect variegation within the expressing cell types. PMID- 12727888 TI - Structural basis for the recognition of ldb1 by the N-terminal LIM domains of LMO2 and LMO4. AB - LMO2 and LMO4 are members of a small family of nuclear transcriptional regulators that are important for both normal development and disease processes. LMO2 is essential for hemopoiesis and angiogenesis, and inappropriate overexpression of this protein leads to T-cell leukemias. LMO4 is developmentally regulated in the mammary gland and has been implicated in breast oncogenesis. Both proteins comprise two tandemly repeated LIM domains. LMO2 and LMO4 interact with the ubiquitous nuclear adaptor protein ldb1/NLI/CLIM2, which associates with the LIM domains of LMO and LIM homeodomain proteins via its LIM interaction domain (ldb1 LID). We report the solution structures of two LMO:ldb1 complexes (PDB: 1M3V and 1J2O) and show that ldb1-LID binds to the N-terminal LIM domain (LIM1) of LMO2 and LMO4 in an extended conformation, contributing a third strand to a beta hairpin in LIM1 domains. These findings constitute the first molecular definition of LIM-mediated protein-protein interactions and suggest a mechanism by which ldb1 can bind a variety of LIM domains that share low sequence homology. PMID- 12727889 TI - Unified two-metal mechanism of RNA synthesis and degradation by RNA polymerase. AB - In DNA-dependent RNA polymerases, reactions of RNA synthesis and degradation are performed by the same active center (in contrast to DNA polymerases in which they are separate). We propose a unified catalytic mechanism for multisubunit RNA polymerases based on the analysis of its 3'-5' exonuclease reaction in the context of crystal structure. The active center involves a symmetrical pair of Mg(2+) ions that switch roles in synthesis and degradation. One ion is retained permanently and the other is recruited ad hoc for each act of catalysis. The weakly bound Mg(2+) is stabilized in the active center in different modes depending on the type of reaction: during synthesis by the beta,gamma-phosphates of the incoming substrate; and during hydrolysis by the phosphates of a non-base paired nucleoside triphosphate. The latter mode defines a transient, non-specific nucleoside triphosphate-binding site adjacent to the active center, which may serve as a gateway for polymerization of substrates. PMID- 12727890 TI - Methylation-induced G(2)/M arrest requires a full complement of the mismatch repair protein hMLH1. AB - The mismatch repair (MMR) gene hMLH1 is mutated in approximately 50% of hereditary non-polyposis colon cancers and transcriptionally silenced in approximately 25% of sporadic tumours of the right colon. Cells lacking hMLH1 display microsatellite instability and resistance to killing by methylating agents. In an attempt to study the phenotypic effects of hMLH1 downregulation in greater detail, we designed an isogenic system, in which hMLH1 expression is regulated by doxycycline. We now report that human embryonic kidney 293T cells expressing high amounts of hMLH1 were MMR-proficient and arrested at the G(2)/M cell cycle checkpoint following treatment with the DNA methylating agent N-methyl N'-nitro-N-nitrosoguanidine (MNNG), while cells not expressing hMLH1 displayed a MMR defect and failed to arrest upon MNNG treatment. Interestingly, MMR proficiency was restored even at low hMLH1 concentrations, while checkpoint activation required a full complement of hMLH1. In the MMR-proficient cells, activation of the MNNG-induced G(2)/M checkpoint was accompanied by phosphorylation of p53, but the cell death pathway was p53 independent, as the latter polypeptide is functionally inactivated in these cells by SV40 large T antigen. PMID- 12727891 TI - Functional interaction between PARP-1 and PARP-2 in chromosome stability and embryonic development in mouse. AB - The DNA damage-dependent poly(ADP-ribose) polymerases, PARP-1 and PARP-2, homo- and heterodimerize and are both involved in the base excision repair (BER) pathway. Here, we report that mice carrying a targeted disruption of the PARP-2 gene are sensitive to ionizing radiation. Following alkylating agent treatment, parp-2(-/-)-derived mouse embryonic fibroblasts exhibit increased post replicative genomic instability, G(2)/M accumulation and chromosome mis segregation accompanying kinetochore defects. Moreover, parp-1(-/-)parp-2(-/-) double mutant mice are not viable and die at the onset of gastrulation, demonstrating that the expression of both PARP-1 and PARP-2 and/or DNA-dependent poly(ADP-ribosyl) ation is essential during early embryogenesis. Interestingly, specific female embryonic lethality is observed in parp-1(+/-)parp-2(-/-) mutants at E9.5. Meta phase analyses of E8.5 embryonic fibroblasts highlight a specific instability of the X chromosome in those females, but not in males. Together, these results support the notion that PARP-1 and PARP-2 possess both overlapping and non-redundant functions in the maintenance of genomic stability. PMID- 12727892 TI - CTG repeat instability and size variation timing in DNA repair-deficient mice. AB - Type 1 myotonic dystrophy is caused by the expansion of an unstable CTG repeat in the DMPK gene. We have investigated the molecular mechanisms underlying the CTG repeat instability by crossing transgenic mice carrying >300 unstable CTG repeats in their human chromatin environment with mice knockout for genes involved in various DNA repair pathways: Msh2 (mismatch repair), Rad52 and Rad54 (homologous recombination) and DNA-PKcs (non-homologous end-joining). Genes of the non homologous end-joining and homologous recombination pathways did not seem to affect repeat instability. Only lack of Rad52 led to a slight decrease in expansion range. Unexpectedly, the absence of Msh2 did not result in stabilization of the CTG repeats in our model. Instead, it shifted the instability towards contractions rather than expansions, both in tissues and through generations. Furthermore, we carefully analyzed repeat transmissions with different Msh2 genotypes to determine the timing of intergenerational instability. We found that instability over generations depends not only on parental germinal instability, but also on a second event taking place after fertilization. PMID- 12727893 TI - Non-homologous end joining as an important mutagenic process in cell cycle arrested cells. AB - Resting cells experience mutations without apparent external mutagenic influences. Such DNA replication-independent mutations are suspected to be a consequence of processing of spontaneous DNA lesions. Using experimental systems based on reversions of frameshift alleles in Saccharomyces cerevisiae, we evaluated the impact of defects in DNA double-strand break (DSB) repair on the frequency of replication-independent mutations. The deletion of the genes coding for Ku70 or DNA ligase IV, which are both obligatory constituents of the non homologous end joining (NHEJ) pathway, each resulted in a 50% reduction of replication-independent mutation frequency in haploid cells. Sequencing indicated that typical NHEJ-dependent reversion events are small deletions within mononucleotide repeats, with a remarkable resemblance to DNA polymerase slippage errors. Experiments with diploid and RAD52- or RAD54-deficient strains confirmed that among DSB repair pathways only NHEJ accounts for a considerable fraction of replication-independent frameshift mutations in haploid and diploid NHEJ non repressed cells. Thus our results provide evidence that G(0) cells with unrepressed NHEJ capacity pay for a large-scale chromosomal stability with an increased frequency of small-scale mutations, a finding of potential relevance for carcinogenesis. PMID- 12727894 TI - Monopolar spindle attachment of sister chromatids is ensured by two distinct mechanisms at the first meiotic division in fission yeast. AB - At meiosis I, sister chromatids attach to the same spindle pole (i.e. monopolar attachment). Mechanisms establishing monopolar attachment remain largely unknown. In the fission yeast Schizosaccharomyces pombe, monopolar attachment is established in haploid cells, indicating that homologous chromosomes are dispensable for its establishment. This monopolar attachment requires both mating pheromone signaling and inactivation of Pat1 kinase (a key negative regulator of meiosis). It also requires the meiotic cohesin factor Rec8 but not the recombination factor Rec12. In contrast, in diploid cells, monopolar attachment is established by Pat1 inactivation alone, and does not require mating pheromone signaling. Furthermore, monopolar attachment requires Rec12 in addition to Rec8. These results indicate that monopolar attachment of sister chromatids can be established by two distinct mechanisms in S.pombe, one that is pheromone dependent and recombination independent, and a second that is pheromone independent and recombination dependent. We propose that co-operation of these two mechanisms generates the high fidelity of monopolar attachment. PMID- 12727895 TI - The integral membrane protein p16.7 organizes in vivo phi29 DNA replication through interaction with both the terminal protein and ssDNA. AB - Remarkably little is known about the in vivo organization of membrane-associated prokaryotic DNA replication or the proteins involved. We have studied this fundamental process using the Bacillus subtilis phage phi29 as a model system. Previously, we demonstrated that the phi29-encoded dimeric integral membrane protein p16.7 binds to ssDNA and is involved in the organization of membrane associated phi29 DNA replication. Here we demonstrate that p16.7 forms multimers, both in vitro and in vivo, and interacts with the phi29 terminal protein. In addition, we show that in vitro multimerization is enhanced in the presence of ssDNA and that the C-terminal region of p16.7 is required for multimerization but not for ssDNA binding or interaction with the terminal protein. Moreover, we provide evidence that the ability of p16.7 to form multimers is crucial for its ssDNA-binding mode. These and previous results indicate that p16.7 encompasses four distinct modules. An integrated model of the structural and functional domains of p16.7 in relation to the organization of in vivo phi29 DNA replication is presented. PMID- 12727896 TI - Comparative analysis of vertebrate dystrophin loci indicate intron gigantism as a common feature. AB - The human DMD gene is the largest known to date, spanning > 2000 kb on the X chromosome. The gene size is mainly accounted for by huge intronic regions. We sequenced 190 kb of Fugu rubripes (pufferfish) genomic DNA corresponding to the complete dystrophin gene (FrDMD) and provide the first report of gene structure and sequence comparison among dystrophin genomic sequences from different vertebrate organisms. Almost all intron positions and phases are conserved between FrDMD and its mammalian counterparts, and the predicted protein product of the Fugu gene displays 55% identity and 71% similarity to human dystrophin. In analogy to the human gene, FrDMD presents several-fold longer than average intronic regions. Analysis of intron sequences of the human and murine genes revealed that they are extremely conserved in size and that a similar fraction of total intron length is represented by repetitive elements; moreover, our data indicate that intron expansion through repeat accumulation in the two orthologs is the result of independent insertional events. The hypothesis that intron length might be functionally relevant to the DMD gene regulation is proposed and substantiated by the finding that dystrophin intron gigantism is common to the three vertebrate genes. PMID- 12727897 TI - Genome-wide in silico identification of transcriptional regulators controlling the cell cycle in human cells. AB - Dissection of regulatory networks that control gene transcription is one of the greatest challenges of functional genomics. Using human genomic sequences, models for binding sites of known transcription factors, and gene expression data, we demonstrate that the reverse engineering approach, which infers regulatory mechanisms from gene expression patterns, can reveal transcriptional networks in human cells. To date, such methodologies were successfully demonstrated only in prokaryotes and low eukaryotes. We developed computational methods for identifying putative binding sites of transcription factors and for evaluating the statistical significance of their prevalence in a given set of promoters. Focusing on transcriptional mechanisms that control cell cycle progression, our computational analyses revealed eight transcription factors whose binding sites are significantly overrepresented in promoters of genes whose expression is cell cycle-dependent. The enrichment of some of these factors is specific to certain phases of the cell cycle. In addition, several pairs of these transcription factors show a significant co-occurrence rate in cell-cycle-regulated promoters. Each such pair indicates functional cooperation between its members in regulating the transcriptional program associated with cell cycle progression. The methods presented here are general and can be applied to the analysis of transcriptional networks controlling any biological process. PMID- 12727898 TI - Complex evolution of 7E olfactory receptor genes in segmental duplications. AB - Large segmental duplications (SDs) constitute at least 3.6% of the human genome and have increased its size, complexity, and diversity. SDs can mediate ectopic sequence exchange resulting in gross chromosomal rearrangements that could contribute to speciation and disease. We have identified and evaluated a subset of human SDs that harbor an 88-member subfamily of olfactory receptor (OR)-like genes called the 7Es. At least 92% of these genes appear to be pseudogenes when compared to other OR genes. The 7E-containing SDs (7E SDs) have duplicated to at least 35 regions of the genome via intra- and interchromosomal duplication events. In contrast to many human SDs, the 7E SDs are not biased towards pericentromeric or subtelomeric regions. We find evidence for gene conversion among 7E genes and larger sequence exchange between 7E SDs, supporting the hypothesis that long, highly similar stretches of DNA facilitate ectopic interactions. The complex structure and history of the 7E SDs necessitates extension of the current model of large-scale DNA duplication. Despite their appearance as pseudogenes, some 7E genes exhibit a signature of purifying selection, and at least one 7E gene is expressed. PMID- 12727899 TI - Parallel evolution by gene duplication in the genomes of two unicellular fungi. AB - Phylogenetic analysis of conserved gene families in fission yeast Schizosaccharomyces pombe and brewer's yeast Saccharomyces cerevisiae showed that gene duplications have occurred independently in the same families in each of these two lineages to a far greater extent than expected by chance. These species represent distinct lineages of the phylum Ascomycota that independently evolved a "yeast" life cycle with a unicellular thallus that reproduces by budding, and many of the genes that have duplicated independently in the two lineages are known to be involved in crucial aspects of this life cycle. Parallel gene duplication thus appears to have played a role in the independent origin of similar adaptations in the two species. The results indicate that using phylogenetic analysis to test for parallel gene duplication in different species may help in identifying genes responsible for similar but independently evolved adaptations. PMID- 12727900 TI - Retroposed copies of the HMG genes: a window to genome dynamics. AB - Retroposed copies (RPCs) of genes are functional (intronless paralogs) or nonfunctional (processed pseudogenes) copies derived from mRNA through a process of retrotransposition. Previous studies found that gene families involved in mRNA translation or nuclear function were more likely to have large numbers of RPCs. Here we characterize RPCs of the few families coding for the abundant high mobility-group (HMG) proteins in humans. Using an algorithm we developed, we identified and studied 219 HMG RPCs. For slightly more than 10% of these RPCs, we found evidence indicating expression. Furthermore, eight of these are potentially new members of the HMG families of proteins. For three RPCs, the evidence indicated expression as part of other transcripts; in all of these, we found the presence of alternative splicing or multiple polyadenylation signals. RPC distribution among the HMGs was not even, with 33-65 each for HMGB1, HMGB3, HMGN1, and HMGN2, and 0-6 each for HMGA1, HMGA2, HMGB2, and HMGN3. Analysis of the sequences flanking the RPCs revealed that the junction between the target site duplications and the 5'-flanking sequences exhibited the same TT/AAAA consensus found for the L1 endonuclease, supporting an L1-mediated retrotransposition mechanism. Finally, because our algorithm included aligning RPC flanking sequences with the corresponding HMG genomic sequence, we were able to identify transcribed regions of HMG genes that were not part of the published mRNA sequences. PMID- 12727901 TI - Quantitative estimates of sequence divergence for comparative analyses of mammalian genomes. AB - Comparative sequence analyses on a collection of carefully chosen mammalian genomes could facilitate identification of functional elements within the human genome and allow quantification of evolutionary constraint at the single nucleotide level. High-resolution quantification would be informative for determining the distribution of important positions within functional elements and for evaluating the relative importance of nucleotide sites that carry single nucleotide polymorphisms (SNPs). Because the level of resolution in comparative sequence analyses is a direct function of sequence diversity, we propose that the information content of a candidate mammalian genome be defined as the sequence divergence it would add relative to already-sequenced genomes. We show that reliable estimates of genomic sequence divergence can be obtained from small genomic regions. On the basis of a multiple sequence alignment of approximately 1.4 megabases each from eight mammals, we generate such estimates for five unsequenced mammals. Estimates of the neutral divergence in these data suggest that a small number of diverse mammalian genomes in addition to human, mouse, and rat would allow single nucleotide resolution in comparative sequence analyses. PMID- 12727903 TI - Selection on human genes as revealed by comparisons to chimpanzee cDNA. AB - To better understand the evolutionary forces that affect human genes, we sequenced 5055 expressed sequence tags from the chimpanzee and compared them to their human counterparts. In conjunction with intergenic chimpanzee DNA sequences and data on human single-nucleotide polymorphisms in the genes studied, this allows us to gauge the extent to which selection affects human genes at a genome wide scale. The comparison to intergenic DNA sequences indicates that about 39% of silent sites in protein-coding regions are deleterious and subject to negative selection. Further, when the divergence between human and chimpanzee is compared with the extent of nucleotide polymorphisms among humans in the same sequences, there is significantly higher divergence in the 5' untranslated regions (UTRs) but not in other parts of the transcript. This indicates that positive selection may have had a considerable influence on 5'UTRs. The dinucleotide CG (CpG) also exhibits a different substitution pattern within 5'UTRs as compared with other parts of the genome. PMID- 12727902 TI - Genome size evolution in pufferfish: a comparative analysis of diodontid and tetraodontid pufferfish genomes. AB - Smooth pufferfish of the family Tetraodontidae have the smallest vertebrate genomes yet measured. They have a haploid genome size of approximately 400 million bp (Mb), which is almost eight times smaller than the human genome. Given that spiny pufferfish from the sister family Diodontidae and a fish from the outgroup Molidae have genomes twice as large as smooth puffers, it appears that the genome size of smooth puffers has contracted in the last 50-70 million years since their divergence from the spiny puffers. Here we use renaturation kinetics to compare the repetitive nature of the smooth and spiny puffer genomes. We also estimate the rates of small (<400 bp) insertions and deletions in smooth and spiny puffers using defunct non-LTR retrotransposons. We find a significantly greater abundance of a transposon-like repetitive DNA class in spiny puffers relative to smooth puffers, in addition to nearly identical indel rates. We comment on the role that large insertions may play in the evolution of genome size in these two groups. PMID- 12727904 TI - Neutral substitutions occur at a faster rate in exons than in noncoding DNA in primate genomes. AB - Point mutation rates in exons (synonymous sites) and noncoding (introns and intergenic) regions are generally assumed to be the same. However, comparative sequence analyses of synonymous substitutions in exons (81 genes) and that of long intergenic fragments (141.3 kbp) of human and chimpanzee genomes reveal a 30%-60% higher mutation rate in exons than in noncoding DNA. We propose a differential CpG content hypothesis to explain this fundamental, and seemingly unintuitive, pattern. We find that the increased exonic rate is the result of the relative overabundance of synonymous sites involved in CpG dinucleotides, as the evolutionary divergence in non-CpG sites is similar in noncoding DNA and synonymous sites of exons. Expectations and predictions of our hypothesis are confirmed in comparisons involving more distantly related species, including human-orangutan, human-baboon, and human-macaque. Our results suggest an underlying mechanism for higher mutation rate in GC-rich genomic regions, predict nonlinear accumulation of mutations in pseudogenes over time, and provide a possible explanation for the observed higher diversity of single nucleotide polymorphisms (SNPs) in the synonymous sites of exons compared to the noncoding regions. PMID- 12727906 TI - Fidelity of the methylation pattern and its variation in the genome. AB - The methylated or unmethylated status of a CpG site is copied faithfully from parental DNA to daughter DNA, and functions as a cellular memory. However, no information is available for the fidelity of methylation pattern in unmethylated CpG islands (CGIs) or its variation in the genome. Here, we determined the methylation status of each CpG site on each DNA molecule obtained from clonal populations of normal human mammary epithelial cells. Methylation pattern error rates (MPERs) were calculated based upon the deviation from the methylation patterns that should be obtained if the cells had 100% fidelity in replicating the methylation pattern. Unmethylated CGIs in the promoter regions of five genes showed MPERs of 0.018-0.032 errors/site/21.6 generations, and the fidelity of methylation pattern was calculated as 99.85%-99.92%/site/generation. In contrast, unmethylated CGIs outside the promoter regions showed MPERs more than twice as high (P < 0.01). Methylated regions, including a CGI in the MAGE-A3 promoter and DMR of the H19 gene, showed much lower MPERs than unmethylated CGIs. These showed that errors in methylation pattern were mainly due to de novo methylations in unmethylated regions. The differential MPERs even among unmethylated CGIs indicated that a promoter-specific protection mechanism(s) from de novo methylation was present. PMID- 12727905 TI - Haplotype structure, LD blocks, and uneven recombination within the LRP5 gene. AB - Patterns of linkage disequilibrium (LD) in the human genome are beginning to be characterized, with a paucity of haplotype diversity in "LD blocks," interspersed by apparent "hot spots" of recombination. Previously, we cloned and physically characterized the low-density lipoprotein-receptor-related protein 5 (LRP5) gene. Here, we have extensively analysed both LRP5 and its flanking three genes, spanning 269 kb, for single nucleotide polymorphisms (SNPs), and we present a comprehensive SNP map comprising 95 polymorphisms. Analysis revealed high levels of recombination across LRP5, including a hot-spot region from intron 1 to intron 7 of LRP5, where there are 109 recombinants/Mb (4882 meioses), in contrast to flanking regions of 14.6 recombinants/Mb. This region of high recombination could be delineated into three to four hot spots, one within a 601-bp interval. For LRP5, three haplotype blocks were identified, flanked by the hot spots. Each LD block comprised over 80% common haplotypes, concurring with a previous study of 14 genes that showed that common haplotypes account for at least 80% of all haplotypes. The identification of hot spots in between these LD blocks provides additional evidence that LD blocks are separated by areas of higher recombination. PMID- 12727908 TI - DASH-2: flexible, low-cost, and high-throughput SNP genotyping by dynamic allele specific hybridization on membrane arrays. AB - Genotyping technologies need to be continually improved in terms of their flexibility, cost-efficiency, and throughput, to push forward genome variation analysis. To this end, we have leveraged the inherent simplicity of dynamic allele-specific hybridization (DASH) and coupled it to recent innovations of centrifugal arrays and iFRET. We have thereby created a new genotyping platform we term DASH-2, which we demonstrate and evaluate in this report. The system is highly flexible in many ways (any plate format, PCR multiplexing, serial and parallel array processing, spectral-multiplexing of hybridization probes), thus supporting a wide range of application scales and objectives. Precision is demonstrated to be in the range 99.8-100%, and assay costs are 0.05 USD or less per genotype assignment. DASH-2 thus provides a powerful new alternative for genotyping practice, which can be used without the need for expensive robotics support. PMID- 12727909 TI - DNA analysis by fluorescence quenching detection. AB - The analysis of human genetic variations such as single nucleotide polymorphisms (SNPs) has great applications in genome-wide association studies of complex genetic traits. We have developed an SNP genotyping method based on the primer extension assay with fluorescence quenching as the detection. The template directed dye-terminator incorporation with fluorescence quenching detection (FQ TDI) assay is based on the observation that the intensity of fluorescent dye R110 and R6G-labeled acycloterminators is universally quenched once they are incorporated onto a DNA oligonucleotide primer. By comparing the rate of fluorescence quenching of the two allelic dyes in real time, we have extended this method for allele frequency estimation of SNPs in pooled DNA samples. The kinetic FQ-TDI assay is highly accurate and reproducible both in genotyping and in allele frequency estimation. Allele frequencies estimated by the kinetic FQ TDI assay correlated well with known allele frequencies, with an r(2) value of 0.993. Applying this strategy to large-scale studies will greatly reduce the time and cost for genotyping hundreds and thousands of SNP markers between affected and control populations. PMID- 12727907 TI - Discovering novel cis-regulatory motifs using functional networks. AB - We combined functional information such as protein-protein interactions or metabolic networks with genome information in Saccharomyces cerevisiae to predict cis-regulatory motifs in the upstream region of genes. We developed a new scoring metric combining these two information sources and used this metric in motif discovery. To estimate the statistical significance of this metric, we used brute force randomization, which shows a consistent well-behaved trend. In contrast, real data showed complex nonrandom behavior. With conservative parameters we were able to find 42 degenerate motifs (that touch 40% of yeast genes) based on 647 original patterns, five of which are well known. Some of these motifs also show limited spatial position in the promoter, indicative of a true motif. We also tested the metric on other known motifs and show that this metric is a good discriminator of real motifs. As well as a pragmatic motif discovery method, with many applications beyond this work, these results also show that interacting proteins are often coordinated at the level of transcription, even in the absence of obvious coregulation in gene expression data sets. PMID- 12727910 TI - Software for automated analysis of DNA fingerprinting gels. AB - Here we describe software tools for the automated detection of DNA restriction fragments resolved on agarose fingerprinting gels. We present a mathematical model for the location and shape of the restriction fragments as a function of fragment size, with model parameters determined empirically from "marker" lanes containing molecular size standards. Automated identification of restriction fragments involves several steps, including: image preprocessing, to put the data in a form consistent with a linear model; marker lane analysis, for determination of the model parameters; and data lane analysis, a procedure for detecting restriction fragment multiplets while simultaneously determining the amplitude curve that describes restriction fragment amplitude as a function of mobility. In validation experiments conducted on fingerprinted and sequenced Bacterial Artificial Chromosome (BAC) clones, sensitivity and specificity of restriction fragment identification exceeded 96% on restriction fragments ranging in size from 600 base pairs (bp) to 30,000 bp. The integrated suite of software tools, written in MATLAB and collectively called BandLeader, is in use at the BC Cancer Agency Genome Sciences Centre (GSC) and the Washington University Genome Sequencing Center, and has been provided to the Wellcome Trust Sanger Institute and the Whitehead Institute. Employed in a production mode at the GSC, BandLeader has been used to perform automated restriction fragment identification for more than 850,000 BAC clones for mouse, rat, bovine, and poplar fingerprint mapping projects. PMID- 12727911 TI - Selecting open reading frames from DNA. AB - We describe a method to select DNA encoding functional open reading frames (ORFs) from noncoding DNA within the context of a specific vector. Phage display has been used as an example, but any system requiring DNA encoding protein fragments, for example, the yeast two-hybrid system, could be used. By cloning DNA fragments upstream of a fusion gene, consisting of the beta-lactamase gene flanked by lox recombination sites, which is, in turn, upstream of gene 3 from fd phage, only those clones containing DNA fragments encoding ORFs confer ampicillin resistance and survive. After selection, the beta-lactamase gene can be removed by Cre recombinase, leaving a standard phage display vector with ORFs fused to gene 3. This vector has been tested on a plasmid containing tissue transglutaminase. All surviving clones analyzed by sequencing were found to contain ORFs, of which 83% were localized to known genes, and at least 80% produced immunologically detectable polypeptides. Use of a specific anti-tTG monoclonal antibody allowed the identification of clones containing the correct epitope. This approach could be applicable to the efficient selection of random ORFs representing the coding potential of whole organisms, and their subsequent downstream use in a number of different systems. PMID- 12727912 TI - In situ-synthesized novel microarray optimized for mouse stem cell and early developmental expression profiling. AB - Applications of microarray technologies to mouse embryology/genetics have been limited, due to the nonavailability of microarrays containing large numbers of embryonic genes and the gap between microgram quantities of RNA required by typical microarray methods and the miniscule amounts of tissue available to researchers. To overcome these problems, we have developed a microarray platform containing in situ-synthesized 60-mer oligonucleotide probes representing approximately 22,000 unique mouse transcripts, assembled primarily from sequences of stem cell and embryo cDNA libraries. We have optimized RNA labeling protocols and experimental designs to use as little as 2 ng total RNA reliably and reproducibly. At least 98% of the probes contained in the microarray correspond to clones in our publicly available collections, making cDNAs readily available for further experimentation on genes of interest. These characteristics, combined with the ability to profile very small samples, make this system a resource for stem cell and embryogenomics research. PMID- 12727915 TI - A friend within the heart: natriuretic peptide receptor signaling. PMID- 12727914 TI - Antimicrobial resistance: the example of Staphylococcus aureus. PMID- 12727916 TI - Double target for tumor mass destruction. PMID- 12727917 TI - Peptide-based treatment for autoimmune diseases: learning how to handle a double edged sword. PMID- 12727918 TI - Tissue glycogen content and glucose intolerance. PMID- 12727913 TI - Analysis of 5'-end sequences of chimpanzee cDNAs. AB - We constructed full-length enriched cDNA libraries from chimpanzee brain, skin, and liver tissues by the oligo-capping method to establish a database of sequences of chimpanzee genes. Randomly selected clones from the libraries were subjected to one-pass sequencing from their 5'-ends. As a result, we collected 6813 chimpanzee cDNA sequences longer than 400 bp. Homology search against human mRNA sequences (RefSeq mRNAs) revealed that our collection included sequences of 1652 putative chimpanzee genes. In order to calculate the sequence identity between human and chimpanzee homologs, we constructed 5'-end consensus sequences of 226 chimpanzee genes by aligning at least three sequences for individual genes. Sequence identity was estimated by comparing these consensus sequences and the corresponding sequences of their human homologs. The average sequence identity of the 5'-end cDNAs was 99.30%. Those of the 5'-UTRs and CDSs were 98.79% and 99.42%, respectively. The results confirmed that human and chimpanzee genes are highly conserved at the nucleotide level. As for amino acids, the average sequence identity was 99.44%. The average synonymous (K(S)) and nonsynonymous (K(A)) divergences were estimated to be 1.33% and 0.28%, respectively. PMID- 12727919 TI - Connecting the dots from Toll-like receptors to innate immune cells and inflammatory bowel disease. PMID- 12727920 TI - Benefits of targeting both pericytes and endothelial cells in the tumor vasculature with kinase inhibitors. AB - Functions of receptor tyrosine kinases implicated in angiogenesis were pharmacologically impaired in a mouse model of pancreatic islet cancer. An inhibitor targeting VEGFRs in endothelial cells (SU5416) is effective against early-stage angiogenic lesions, but not large, well-vascularized tumors. In contrast, a kinase inhibitor incorporating selectivity for PDGFRs (SU6668) is shown to block further growth of end-stage tumors, eliciting detachment of pericytes and disruption of tumor vascularity. Importantly, PDGFRs were expressed only in perivascular cells of this tumor type, suggesting that PDGFR(+) pericytes in tumors present a complimentary target to endothelial cells for efficacious antiangiogenic therapy. Therapeutic regimes combining the two kinase inhibitors (SU5416 and SU6668) were more efficacious against all stages of islet carcinogenesis than either single agent. Combination of the VEGFR inhibitor with another distinctive kinase inhibitor targeting PDGFR activity (Gleevec) was also able to regress late-stage tumors. Thus, combinatorial targeting of receptor tyrosine kinases shows promise for treating multiple stages in tumorigenesis, most notably the often-intractable late-stage solid tumor. PMID- 12727922 TI - Estrogen modulates cutaneous wound healing by downregulating macrophage migration inhibitory factor. AB - Characteristic of both chronic wounds and acute wounds that fail to heal are excessive leukocytosis and reduced matrix deposition. Estrogen is a major regulator of wound repair that can reverse age-related impaired wound healing in human and animal models, characterized by a dampened inflammatory response and increased matrix deposited at the wound site. Macrophage migration inhibitory factor (MIF) is a candidate proinflammatory cytokine involved in the hormonal regulation of inflammation. We demonstrate that MIF is upregulated in a distinct spatial and temporal pattern during wound healing and its expression is markedly elevated in wounds of estrogen-deficient mice as compared with intact animals. Wound-healing studies in mice rendered null for the MIF gene have demonstrated that in the absence of MIF, the excessive inflammation and delayed-healing phenotype associated with reduced estrogen is reversed. Moreover, in vitro assays have shown a striking estrogen-mediated decrease in MIF production by activated murine macrophages, a process involving the estrogen receptor. We suggest that estrogen inhibits the local inflammatory response by downregulating MIF, suggesting a specific target for future therapeutic intervention in impaired wound-healing states. PMID- 12727923 TI - A functional androgen receptor is not sufficient to allow estradiol to protect bone after gonadectomy in estradiol receptor-deficient mice. AB - Although the role of estradiol in maintaining bone mass is well established, the relative contributions of the estradiol receptors ERalpha and ERbeta and of the androgen receptor (AR) remain controversial. To determine the role of ERalpha mediated, ERbeta-mediated, and non-ER-mediated mechanisms in maintaining bone mass, gonadectomy and estradiol treatment were studied in ER-knockout mice. Estradiol treatment of ovariectomized ERalphabeta(-/-) mice failed to prevent bone loss, precluding significant effects of estradiol on bone through non-ER signaling pathways. In contrast, estradiol prevented ovariectomy-induced bone loss in ERbeta(-/-) mice, as in WT males and females, indicating that ERalpha is the major mediator of estradiol effects in bone. No response of bone to estradiol was detected in orchidectomized ERalpha(-/-) mice, suggesting estradiol cannot protect bone mass via the AR in vivo. In contrast to female ERalphabeta(-/-) and male ERalpha(-/-) mice, female ERalpha(-/-) mice were partially protected against ovariectomy-induced bone loss by estradiol, confirming that ERbeta mediates estradiol effects in bone, but only in females and with a lower efficacy than ERalpha. We conclude that ERalpha is the main effector of estradiol's protective function in bone in both male and female mice, and that, in its absence, AR is not sufficient to mediate this response. PMID- 12727921 TI - Toll-like receptor-dependent production of IL-12p40 causes chronic enterocolitis in myeloid cell-specific Stat3-deficient mice. AB - Stat3 plays an essential role in IL-10 signaling pathways. A myeloid cell specific deletion of Stat3 resulted in inflammatory cytokine production and development of chronic enterocolitis with enhanced Th1 responses in mice. In this study, we analyzed the mechanism by which a Stat3 deficiency in myeloid cells led to the induction of chronic enterocolitis in vivo. Even in the absence of Stat1, which is essential for IFN-gamma signaling pathways, Stat3 mutant mice developed chronic enterocolitis. TNF-alpha/Stat3 double-mutant mice developed severe chronic enterocolitis with enhanced Th1 cell development. IL-12p40/Stat3 double mutant mice, however, showed normal Th1 responses and no inflammatory change in the colon. RAG2/Stat3 double-mutant mice did not develop enterocolitis, either. These findings indicate that overproduction of IL-12p40, which induces potent Th1 responses, is essential for the development of chronic enterocolitis in Stat3 mutant mice. Furthermore, enterocolitis was significantly improved and IFN-gamma production by T cells was reduced in TLR4/Stat3 double-mutant mice, indicating that TLR4-mediated recognition of microbial components triggers aberrant IL-12p40 production by myeloid cells, leading to the development of enterocolitis. Thus, this study clearly established a sequential innate and acquired immune mechanism for the development of Th1-dependent enterocolitis. PMID- 12727925 TI - Vasoactive intestinal peptide as a new drug for treatment of primary pulmonary hypertension. AB - Primary pulmonary hypertension is a fatal disease causing progressive right heart failure within 3 years after diagnosis. We describe a new concept for treatment of the disease using vasoactive intestinal peptide, a neuropeptide primarily functioning as a neurotransmitter that acts as a potent systemic and pulmonary vasodilator. Our rationale is based on the finding of a deficiency of the peptide in serum and lung tissue of patients with primary pulmonary hypertension, as evidenced by radioimmunoassay and immunohistochemistry. The relevance of this finding is underlined by an upregulation of corresponding receptor sites as shown by Northern blot analysis, Western blot analysis, and immunological techniques. Consequently, the substitution with the hormone results in substantial improvement of hemodynamic and prognostic parameters of the disease without side effects. It decreased the mean pulmonary artery pressure in our eight study patients, increased cardiac output, and mixed venous oxygen saturation. Our data provide enough proof for further investigation of vasoactive intestinal peptide and its role in primary pulmonary hypertension. PMID- 12727924 TI - Anti-inflammatory properties of the mu opioid receptor support its use in the treatment of colon inflammation. AB - The physiologic role of the mu opioid receptor (MOR) in gut nociception, motility, and secretion is well established. To evaluate whether MOR may also be involved in controlling gut inflammation, we first showed that subcutaneous administration of selective peripheral MOR agonists, named DALDA and DAMGO, significantly reduces inflammation in two experimental models of colitis induced by administration of 2,4,6-trinitrobenzene sulfonic acid (TNBS) or peripheral expansion of CD4(+) T cells in mice. This therapeutic effect was almost completely abolished by concomitant administration of the opioid antagonist naloxone. Evidence of a genetic role for MOR in the control of gut inflammation was provided by showing that MOR-deficient mice were highly susceptible to colon inflammation, with a 50% mortality rate occurring 3 days after TNBS administration. The mechanistic basis of these observations suggests that the anti-inflammatory effects of MOR in the colon are mediated through the regulation of cytokine production and T cell proliferation, two important immunologic events required for the development of colon inflammation in mice and patients with inflammatory bowel disease (IBD). These data provide evidence that MOR plays a role in the control of gut inflammation and suggest that MOR agonists might be new therapeutic molecules in IBD. PMID- 12727927 TI - Transfer of hematopoietic stem cells encoding autoantigen prevents autoimmune diabetes. AB - Bone marrow or hematopoietic stem cell transplantation is a potential treatment for autoimmune disease. The clinical application of this approach is, however, limited by the risks associated with allogeneic transplantation. In contrast, syngeneic transplantation would be safe and have wide clinical application. Because T cell tolerance can be induced by presenting antigen on resting antigen presenting cells (APCs), we reasoned that hematopoietic stem cells engineered to express autoantigen in resting APCs could be used to prevent autoimmune disease. Proinsulin is a major autoantigen associated with pancreatic beta cell destruction in humans with type 1 diabetes (T1D) and in autoimmune NOD mice. Here, we demonstrate that syngeneic transplantation of hematopoietic stem cells encoding proinsulin transgenically targeted to APCs totally prevents the development of spontaneous autoimmune diabetes in NOD mice. This antigen-specific immunotherapeutic strategy could be applied to prevent T1D and other autoimmune diseases in humans. PMID- 12727926 TI - Induction of immune tolerance to coagulation factor IX antigen by in vivo hepatic gene transfer. AB - Gene replacement therapy is an attractive approach for treatment of genetic disease, but may be complicated by the risk of a neutralizing immune response to the therapeutic gene product. There are examples of humoral and cellular immune responses against the transgene product as well as absence of such responses, depending on vector design and the underlying mutation in the dysfunctional gene. It has been unclear, however, whether transgene expression can induce tolerance to the therapeutic antigen. Here, we demonstrate induction of immune tolerance to a secreted human coagulation factor IX (hF.IX) antigen by adeno-associated viral gene transfer to the liver. Tolerized mice showed absence of anti-hF.IX and substantially reduced in vitro T cell responses after immunization with hF.IX in adjuvant. Tolerance induction was antigen specific, affected a broad range of Th cell subsets, and was favored by higher levels of transgene expression as determined by promoter strength, vector dose, and mouse strain. Hepatocyte derived hF.IX expression induced regulatory CD4(+) T cells that can suppress anti hF.IX formation after adoptive transfer. With a strain-dependent rate of success, tolerance to murine F.IX was induced in mice with a large F.IX gene deletion, supporting the relevance of these data for treatment of hemophilia B and other genetic diseases. PMID- 12727928 TI - Disabling an integral CTL epitope allows suppression of autoimmune diabetes by intranasal proinsulin peptide. AB - Insulin is a major target of the autoimmune response associated with destruction of pancreatic beta cells in type 1 diabetes. A peptide that spans the junction of the insulin B chain and the connecting (C) peptide in proinsulin has been reported to stimulate T cells from humans at risk for type 1 diabetes and autoimmune diabetes-prone NOD mice. Here we show that proinsulin B24-C36 peptide binds to I-A(g7), the MHC class II molecule of the NOD mouse, and, after intranasal administration, induces regulatory CD4(+) T cells that, in the absence of CD8(+) T cells, block the adoptive transfer of diabetes. Curiously, however, intranasal B24-C36 did not inhibit development of spontaneous diabetes in treated mice. We then determined that B24-C36, and its core sequence B25-C34, bind to K(d), the NOD mouse MHC class I molecule, and elicit CD8(+) CTLs. When the CD8(+) T lymphocyte epitope was truncated at the C34 valine anchor residue for binding to K(d), the residual CD4(+) T cell epitope, B24-C32/33, significantly inhibited diabetes development after a single intranasal dose. This study identifies a novel CTL epitope in proinsulin and demonstrates that the therapeutic potential of a "tolerogenic" autoantigen peptide can be compromised by the presence of an integral CTL epitope. PMID- 12727929 TI - The role of endothelial insulin signaling in the regulation of vascular tone and insulin resistance. AB - Insulin receptors (IRs) on vascular endothelial cells have been suggested to participate in insulin-regulated glucose homeostasis. To directly address the role of insulin action in endothelial function, we have generated a vascular endothelial cell IR knockout (VENIRKO) mouse using the Cre-loxP system. Cultured endothelium of VENIRKO mice exhibited complete rearrangement of the IR gene and a more than 95% decrease in IR mRNA. VENIRKO mice were born at the expected Mendelian ratio, grew normally, were fertile, and exhibited normal patterns of vasculature in the retina and other tissues. Glucose homeostasis under basal condition was comparable in VENIRKO mice. Both eNOS and endothelin-1 mRNA levels, however, were reduced by approximately 30-60% in endothelial cells, aorta, and heart, while vascular EGF expression was maintained at normal levels. Arterial pressure tended to be lower in VENIRKO mice on both low- and high-salt diets, and on a low-salt diet VENIRKO mice showed insulin resistance. Thus, inactivation of the IR on endothelial cell has no major consequences on vascular development or glucose homeostasis under basal conditions, but alters expression of vasoactive mediators and may play a role in maintaining vascular tone and regulation of insulin sensitivity to dietary salt intake. PMID- 12727930 TI - PPAR-gamma receptor ligands: novel therapy for pituitary adenomas. AB - Pituitary tumors cause considerable morbidity due to local invasion, hypopituitarism, or hormone hypersecretion. In many cases, no suitable drug therapies are available, and surgical excision is currently the only effective treatment. We show here abundant expression of nuclear hormone receptor PPAR gamma in all of 39 human pituitary tumors. PPAR-gamma activating thiazolidinediones (TZDs) rosiglitazone and troglitazone induced G(0)-G(1) cell cycle arrest and apoptosis in human, rat somatolactotroph, and murine gonadotroph pituitary tumor cells, and suppressed in vitro hormone secretion. In vivo development and growth of murine somatolactotroph and gonadotroph tumors, generated by subcutaneous injection of prolactin-secreting (PRL-secreting) and growth hormone-secreting (GH-secreting) GH3 cells, luteinizing hormone-secreting (LH-secreting) LbetaT2 cells, and alpha-T3 cells, was markedly suppressed in rosiglitazone-treated mice, and serum GH, PRL, and LH levels were attenuated in all treated animals (P < 0.009). These results demonstrate that PPAR-gamma is an important molecular target in pituitary adenoma cells and PPAR-gamma ligands inhibit tumor cell growth and GH, PRL, and LH secretion in vitro and in vivo. TZDs are proposed as novel oral medications for managing pituitary tumors. PMID- 12727931 TI - Second-site mutation in the Wiskott-Aldrich syndrome (WAS) protein gene causes somatic mosaicism in two WAS siblings. AB - Revertant mosaicism due to true back mutations or second-site mutations has been identified in several inherited disorders. The occurrence of revertants is considered rare, and the underlying genetic mechanisms remain mostly unknown. Here we describe somatic mosaicism in two brothers affected with Wiskott-Aldrich syndrome (WAS). The original mutation causing disease in this family is a single base insertion (1305insG) in the WAS protein (WASP) gene, which results in frameshift and abrogates protein expression. Both patients, however, showed expression of WASP in a fraction of their T cells that were demonstrated to carry a second-site mutation causing the deletion of 19 nucleotides from nucleotide 1299 to 1316. This deletion abrogated the effects of the original mutation and restored the WASP reading frame. In vitro expression studies indicated that mutant protein encoded by the second-site mutation was expressed and functional, since it was able to bind to cellular partners and mediate T cell receptor/CD3 downregulation. These observations were consistent with evidence of in vivo selective advantage of WASP-expressing lymphocytes. Molecular analysis revealed that the sequence surrounding the deletion contained two 4-bp direct repeats and that a hairpin structure could be formed by five GC pairs within the deleted fragment. These findings strongly suggest that slipped mispairing was the cause of this second-site mutation and that selective accumulation of WASP-expressing T lymphocytes led to revertant mosaicism in these patients. PMID- 12727932 TI - Pressure-independent cardiac hypertrophy in mice with cardiomyocyte-restricted inactivation of the atrial natriuretic peptide receptor guanylyl cyclase-A. AB - Cardiac hypertrophy is a common and often lethal complication of arterial hypertension. Atrial natriuretic peptide (ANP) has been postulated to exert local antihypertrophic effects in the heart. Thus, a loss of function of the ANP receptor guanylyl cyclase-A (GC-A) might contribute to the increased propensity to cardiac hypertrophy, although a causative role in vivo has not been definitively demonstrated. To test whether local ANP modulates cardiomyocyte growth, we inactivated the GC-A gene selectively in cardiomyocytes by homologous loxP/Cre-mediated recombination. Thereby we have circumvented the systemic, hypertensive phenotype associated with germline inactivation of GC-A. Mice with cardiomyocyte-restricted GC-A deletion exhibited mild cardiac hypertrophy, markedly increased mRNA expression of cardiac hypertrophy markers such as ANP (fivefold), alpha-skeletal actin (1.7-fold), and beta-myosin heavy chain (twofold), and increased systemic circulating ANP levels. Their blood pressure was 7-10 mmHg below normal, probably because of the elevated systemic levels and endocrine actions of ANP. Furthermore, cardiac hypertrophic responses to aortic constriction were enhanced and accompanied by marked deterioration of cardiac function. This phenotype is consistent with a local function of the ANP/GC-A system to moderate the molecular program of cardiac hypertrophy. PMID- 12727935 TI - A modern medical quandary: polycystic ovary syndrome, insulin resistance, and oral contraceptive pills. PMID- 12727933 TI - The role of falling leptin levels in the neuroendocrine and metabolic adaptation to short-term starvation in healthy men. AB - To elucidate the role of leptin in regulating neuroendocrine and metabolic function during an acute fast, six to eight healthy, lean men were studied under four separate conditions: a baseline fed state and three 72-hour fasting studies with administration of either placebo, low-dose recombinant-methionyl human leptin (r-metHuLeptin), or replacement-dose r-metHuLeptin designed to maintain serum leptin at levels similar to those in the fed state. Replacement-dose r metHuLeptin administered during fasting prevents the starvation-induced changes in the hypothalamic-pituitary-gonadal axis and, in part, the hypothalamic pituitary-thyroid axis and IGF-1 binding capacity in serum. Thus, in normal men, the fall in leptin with fasting may be both necessary and sufficient for the physiologic adaptations of these axes, which require leptin levels above a certain threshold for activation. In contrast to findings in mice, fasting induced changes in the hypothalamic-pituitary-adrenal, renin-aldosterone, and growth hormone-IGF-1 axes as well as fuel utilization may be independent of leptin in humans. The role of leptin in normalizing several starvation-induced neuroendocrine changes may have important implications for the pathophysiology and treatment of eating disorders and obesity. PMID- 12727937 TI - Clinical review 157: Pathophysiology of Graves' ophthalmopathy: the cycle of disease. PMID- 12727936 TI - Clinical review 156: Recombinant human thyrotropin and thyroid cancer management. PMID- 12727934 TI - PTG gene deletion causes impaired glycogen synthesis and developmental insulin resistance. AB - Protein targeting to glycogen (PTG) is a scaffolding protein that targets protein phosphatase 1alpha (PP1alpha) to glycogen, and links it to enzymes involved in glycogen synthesis and degradation. We generated mice that possess a heterozygous deletion of the PTG gene. These mice have reduced glycogen stores in adipose tissue, liver, heart, and skeletal muscle, corresponding with decreased glycogen synthase activity and glycogen synthesis rate. Although young PTG heterozygous mice initially demonstrate normal glucose tolerance, progressive glucose intolerance, hyperinsulinemia, and insulin resistance develop with aging. Insulin resistance in older PTG heterozygous mice correlates with a significant increase in muscle triglyceride content, with a corresponding attenuation of insulin receptor signaling. These data suggest that PTG plays a critical role in glycogen synthesis and is necessary to maintain the appropriate metabolic balance for the partitioning of fuel substrates between glycogen and lipid. PMID- 12727938 TI - Clinical review 158: Beyond radioiodine: a review of potential new therapeutic approaches for thyroid cancer. AB - One of the greatest challenges in the management of patients with follicular cell derived thyroid cancer is the treatment of tumors that progress despite surgery, radioiodine, and T(4) suppression of TSH. As knowledge of thyroid cancer biology improves, the potential exists to develop compounds targeted to treat thyroid cancers that do not respond to traditional therapy. Recently, the development of therapies targeted against specific molecular pathways involved in cancer progression has resulted in dramatic responses in patients with chronic myelogenous leukemia, gastrointestinal stromal tumors, and other cancers. A number of compounds are currently being evaluated in clinical trials that alter pathways involved thyroid cancer, and several of these agents have been tested in thyroid cancer in vitro and in vivo. In this review we will discuss the mechanisms of action and preclinical/clinical data for several of these compounds that have the potential to play an important role in the management of thyroid cancer in the future. PMID- 12727939 TI - Clinical review 159: Human immunodeficiency virus/highly active antiretroviral therapy-associated metabolic syndrome: clinical presentation, pathophysiology, and therapeutic strategies. PMID- 12727940 TI - The future endocrinology workforce. PMID- 12727941 TI - A model to determine workforce needs for endocrinologists in the United States until 2020. AB - The objective of this study was to define the workforce needs for the specialty of endocrinology, diabetes, and metabolism in the United States between 1999 and 2020. An interactive model of factors likely to influence the balance between the supply and demand of endocrinologists during the next 20 yr was constructed. The model used data from a wide range of sources and was developed under the guidance of a panel of experts derived from sponsoring organizations of endocrinologists. We determined current and projected numbers and demographics of endocrinologists in the United States workforce and the anticipated balance between supply and demand from 1999 to 2020. There were 3,623 adult endocrinologists in the workforce in 1999, of which 2,389 (66%) were in office-based practice. Their median age was 49 yr. Both total office visits and services performed by endocrinologists (particularly for diabetes) increased substantially during the 1990s. Waiting time for an initial appointment is presently longer for endocrinologists than for other physicians. Compared with a balanced, largely closed-staff health maintenance organization, the current national supply of endocrinologists is estimated to be 12% lower than demand. The number of endocrinologists entering the market has continuously fallen over the previous 5 yr, from 200 in 1995 to 171 in 1999. Even if this downward trend were abruptly stopped, the model predicts that demand will exceed supply from now until 2020. Whereas this gap narrows from 2000 to 2008 due to projected growth of managed care, it widens thereafter due to the aging of both the population and the endocrine workforce. Inclusion of other factors such as projected real income growth and increased prevalence of age-related endocrine disorders (e.g. diabetes and osteoporosis) further accentuates the deficit. If the number of endocrinologists entering the workforce remains at 1999 levels, demand will continue to exceed supply from now through 2020 for adult endocrinologists, and the gap will widen progressively from 2010 onward. The present analysis indicates that the number of endocrinologists entering the workforce will not be sufficient to meet future demand. These data suggest that steps should be taken to stop the ongoing decline in the number of endocrinologists in training and consideration should be given to actions designed to increase the number of endocrinologists in practice in the years ahead. PMID- 12727943 TI - Functional and nonfunctional adrenocortical tumors demonstrate a high responsiveness to low-dose adrenocorticotropin. AB - Aldosterone-producing adenomas (APAs) demonstrate exquisite sensitivity to endogenous ACTH. We previously showed an ACTH receptor overexpression in APAs compared with the other adrenal tumors. To evaluate the meaning of such findings, we investigated the response of aldosterone, cortisol, and 17OH progesterone (17OHP) to 1 microg ACTH in 42 patients with adrenocortical tumors (23 NHAs, 9 APAs, and 10 CPAs) and 10 normal subjects (C). All 52 subjects were responsive to ACTH, and hormone peak levels were reached at 30 min. The aldosterone peak level was significantly higher in APAs [mean +/- SEM: 84.3 +/- 13.1 ng/dl (2335.1 +/- 362.9 pmol/liter)] than in other tumors and control (C). Cortisol peak levels was higher in CPAs [37.1 +/- 3.9 microg/dl (1023.9 +/- 107.6 nmol/liter)] than in NHAs (P < 0.01), in C (P < 0.01) and in APAs (P = n.s.). 17OHP peak levels were significantly higher in patients with adrenocortical tumors toward C. In summary: 1) low-dose ACTH induces an important stimulation in all tumors, suggesting preservation of high responsiveness to ACTH; 2) this is especially true for aldosterone in APA and could be of primary importance when performing diagnostic tests for hyperaldosteronism; and 3) 17OHP-hyperresponsiveness to low-dose ACTH is the most common alteration both in functional and nonfunctional tumors. PMID- 12727942 TI - Successful pregnancy after bromocriptine therapy in an anovulatory woman complicated with ovarian hyperstimulation caused by follicle-stimulating hormone producing plurihormonal pituitary microadenoma. AB - Gonadotropin-producing pituitary adenomas are extremely rare in reproductive-age women. We report here a case of gonadotroph microadenoma with ovarian hyperstimulation. It was found in a 29-yr-old infertile Japanese woman with enlarged multicystic ovaries. The patient had an elevated basal serum estradiol level (up to 6755 pM, or 1840 pg/ml). Serum FSH and prolactin were mildly elevated (15.4 IU/liter, 1.4 nM or 31.4 ng/ml), whereas LH was low (0.5 IU/liter). The FSH level was paradoxically elevated in response to TRH administration. Dynamic magnetic resonance imaging revealed a pituitary microadenoma. Daily administration of bromocriptine, a dopamine agonist, normalized the ovarian size, and the patient ovulated naturally. She conceived after 3 months of bromocriptine therapy and delivered a normal child. She underwent elective transsphenoidal pituitary surgery, 3 yr after the delivery. Immunostaining of the resected tumor showed that 80% and less than 5% of the tumor cells stained for FSH-beta and prolactin, respectively. Furthermore, RT-PCR suggested that dopamine type 2 receptor was expressed in the adenoma. Gonadotroph microadenoma should be considered in women with spontaneous ovarian hyperstimulation, even if they have no neurological symptoms or marked pituitary enlargement. In such cases, bromocriptine therapy may be an alternative to pituitary surgery. PMID- 12727944 TI - Treatment of type II amiodarone-induced thyrotoxicosis by either iopanoic acid or glucocorticoids: a prospective, randomized study. AB - Amiodarone-induced thyrotoxicosis (AIT) may occur either in the presence of underlying thyroid disease (type I AIT) or in apparently normal thyroid glands (type II AIT). Type II AIT, a destructive thyroiditis, often favorably responds to glucocorticoids. Iopanoic acid (IopAc) is an iodinated cholecystographic agent that inhibits deiodinase activity and reduces the conversion of T(4) toT(3). It has recently been reported that cholecystographic agents restore euthyroidism in patients with type II AIT. We describe the results of a prospective randomized study conducted in 12 patients with type II AIT treated with either iopanoic acid (group A, n = 6) or glucocorticoids (group B, n = 6). Serum free T(3) levels normalized rapidly in both groups after 7 d, from 0.75 +/- 0.20 ng/dl (11.5 +/- 3.1 pmol/liter) to 0.46 +/- 0.10 ng/d (7.1 +/- 1.7 pmol/liter), P < 0.01, and from 0.58 +/- 0.10 ng/dl (9.0 +/- 1.2 pmol/liter) to 0.34 +/- 0.03 ng/dl (5.2 +/- 0.5 pmol/liter), P < 0.003, in groups A and B, respectively (P = NS). Serum free T(4) levels reduced at 6 months in group B [from 2.70 +/- 0.32 ng/dl (35.1 +/- 4.1 pmol/liter) to 1.0 +/- 0.04 ng/dl (13.4 +/- 0.6 pmol/liter), P < 0.0001] but not in group A (from 2.90 +/- 0.6 ng/dl (38.0 +/- 7.5 pmol/liter) to 2.30 +/- 0.4 ng/dl (35.6 +/- 6.1 pmol/liter, P = 0.39; P = 0.005 group B vs. group A). All patients in both groups became euthyroid and had their amiodarone-induced destructive thyroiditis cured as defined by normalization of both serum free T(4) and free T(3) levels, during both drugs therapy. However, patients in group B were cured more rapidly than patients in group A (43 +/- 34 d vs. 221 +/- 111 d, respectively, P < 0.002). This study shows that, albeit both drugs are effective, glucocorticoids are probably the drug of choice for more rapidly curing type II AIT. PMID- 12727945 TI - X chromosome-linked Kallmann syndrome: clinical heterogeneity in three siblings carrying an intragenic deletion of the KAL-1 gene. AB - Kallmann syndrome (KS) is characterized by the association of hypogonadotropic hypogonadism and anosmia. The gene underlying the X chromosome-linked form of the disease, KAL-1, consists of 14 coding exons. It encodes a glycoprotein, anosmin 1, which is involved in the embryonic migration of GnRH-synthesizing neurons and the differentiation of the olfactory bulbs. We describe herein the clinical heterogeneity in three affected brothers who carry a large deletion (exons 3-13) in KAL-1. All three had a history of hypogonadotropic hypogonadism with delayed puberty. Although brain magnetic resonance imaging showed hypoplastic olfactory bulbs in the three siblings, variable degrees of anosmia/hyposmia were shown by olfactometry. In addition, these brothers had different phenotypic anomalies, i.e. unilateral renal aplasia (siblings B and C), high-arched palate (sibling A), brachymetacarpia (sibling A), mirror movements (siblings A and B), and abnormal eye movements (sibling C). Last but not least, sibling A suffered from a severe congenital hearing impairment, a feature that had been reported in KS but had not yet been ascribed unambiguously to the X-linked form of the disease. The variable phenotype, both qualitatively and quantitatively, in this family further emphasizes the role of putative modifier genes, and/or epigenetic factors, in the expressivity of the X-linked KS. PMID- 12727946 TI - Sex-specific impact of congenital hypothyroidism due to thyroid dysgenesis on skeletal maturation in term newborns. AB - Newborns with severe congenital hypothyroidism (often defined by the absence of knee epiphyses at diagnosis) are still at risk of loss of intellectual potential despite early treatment. Although there is no significant sexual dimorphism in the age at appearance and size of the knee epiphyses in normal newborns, it was our clinical impression that these epiphyses were more often absent in hypothyroid newborn males than in affected females. Using the large French database of congenital hypothyroidism, we studied the presence or absence of knee epiphyses at diagnosis, as well as the length of gestation and the birth weight of 1827 term newborns with athyreosis or ectopic thyroid. Boys were twice as likely as girls to have absent epiphyses [odds ratio, 2.1 (95% confidence interval, 1.6-2.7), P < 0.001, after adjustment for etiology, plasma free T(4) concentration, and presence or absence of clinical signs at diagnosis, gestational age and birth weight]. Compared with the general population of French newborns, those with congenital hypothyroidism were more often born after a prolonged gestation (> or =42 wk) and with a high birth weight (9% were above the 95th centile, as opposed to the expected 5%), regardless of sex. We conclude that the impact of congenital hypothyroidism on fetal skeletal maturation is sexually dimorphic. This may result from less efficient conversion of T(4) to T(3) by growth plate chondrocytes in males. PMID- 12727947 TI - Low adiponectin levels in adolescent obesity: a marker of increased intramyocellular lipid accumulation. AB - We examined the impact of adolescent obesity on circulating adiponectin levels and the relationship between adiponectin and insulin sensitivity, intramyocellular (IMCL) lipid content, plasma triglycerides, and free fatty acids. Plasma adiponectin levels were measured in 8 nonobese (percentage fat, 18 +/- 1.8) and 14 obese adolescents (percentage fat, 41 +/- 1.6). Insulin sensitivity was assessed by the euglycemic-hyperinsulinemic clamp. Intramuscular lipid content was quantified using (1)H-nuclear magnetic resonance spectroscopy, and abdominal fat distribution by magnetic resonance imaging. Adiponectin levels were lower in obese adolescents (9.2 +/- 1 microg/ml, P < 0.001) and were positively related to insulin sensitivity in all subjects (r = 0.531, P < 0.02). Strong inverse relationships were found between adiponectin and triglyceride levels (r = -0.80, P < 0.001) and IMCL (r = -0.73, P < 0.001). Triglycerides (partial r(2) = 0.52; P < 0.0002) and IMCL (partial r(2) = 0.10; P < 0.05) were the most significant predictors of adiponectin levels, explaining 62% of the variation. In conclusion, plasma adiponectin levels are reduced in adolescent obesity and related to insulin resistance, independent of total body fat and central adiposity. There is a strong relationship between adiponectin and IMCL lipid content in this pediatric population. The putative modulatory effects of adiponectin on insulin sensitivity may, in part, be mediated via its effects on IMCL lipid content. PMID- 12727948 TI - Insulin-like growth factor I and interleukin-6 contribute synergistically to disability and mortality in older women. AB - The physiology of age-related functional decline is poorly understood, but may involve hormones and inflammation. We hypothesized that older women with both low IGF-I and high IL-6 levels are at high risk for disability and death. We assessed walking speed and disability in 718 women enrolled in the Women's Health and Aging Study I, a 3-yr cohort study with 5-yr mortality follow-up. Women with IGF I levels in the lowest quartile and IL-6 levels in the highest quartile had significantly greater limitation in walking and disability in mobility tasks and instrumental activities of daily living than those with neither risk factor (adjusted odds ratios, 10.77, 5.14, and 3.66). Women with both risk factors were at greater risk for death (adjusted relative risk, 2.10) as well as incident walking limitation, mobility disability, and disability in activities of daily living compared with those with high IGF-I and low IL-6 levels. The combination of low IGF-I and high IL-6 levels confers a high risk for progressive disability and death in older women, suggesting an aggregate effect of dysregulation in endocrine and immune systems. The joint effects of IGF-I and IL-6 may be important targets for treatments to prevent or minimize disability associated with aging. PMID- 12727949 TI - Increase of estrogen dose deteriorates mood during progestin phase in sequential hormonal therapy. AB - Previous studies have indicated that the addition of progestins during sequential hormonal replacement therapy (HRT) causes negative mood and physical symptoms. History of premenstrual syndrome, type of progestin, and dose of progestin have thus far been shown to influence the progestin-induced adverse mood symptoms during HRT. The aim of this study was to compare adverse mood effects of two different doses of estradiol, in combination with a progestin, during postmenopausal HRT. Twenty-eight perimenopausal women were included in this randomized, double-blind, crossover study comparing 2- or 3-mg continuous estradiol, with an addition of 10 mg medroxyprogesterone acetate on d 17-28 during each treatment cycle. The main outcome measures were mood and physical symptoms kept on a daily rating scale. Together with the progestin, the higher dose of estrogen caused significantly more negative mood symptoms than the lower dose. Tension, irritability, and depressed mood were all significantly augmented during the progestin phase of cycles with 3 mg estradiol (P < 0.001). Physical symptoms also increased during the progestin phase of 3-mg estradiol cycles (P < 0.001), whereas positive mood symptoms were less affected. The only positive mood that changed with estrogen dose was friendliness, which decreased during the progestin phase of high estradiol cycles compared with cycles with lower estradiol (P < 0.05). Our conclusion is that an increase of the estrogen dose accentuates negative mood and physical symptoms during the progestin phase of sequential hormonal therapy. PMID- 12727950 TI - Glucose intolerance, insulin resistance, and hyperandrogenemia in first degree relatives of women with polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is associated with hyperinsulinemia, insulin resistance (IR), increased risk of glucose intolerance, and type 2 diabetes. Family studies have indicated a genetic susceptibility to PCOS. The aims of this study were 1) to assess glucose tolerance status, gonadotropins, and androgens in first degree relatives of patients with PCOS; and 2) to assess IR in normal glucose tolerant (NGT) family members. One hundred two family members of 52 patients with PCOS [Mothers(PCOS) (n = 34; mean age, 46.5 yr; mean body mass index (BMI), 28.8 kg/m(2)), Fathers(PCOS) (n = 24; mean age, 50.4 yr; mean BMI, 27.5 kg/m(2)), Sisters(PCOS) (n = 19; mean age, 25.1 yr; mean BMI, 22.9 kg/m(2)), and Brothers(PCOS) (n = 25; mean age, 23.7 yr; mean BMI, 22.5 kg/m(2))] and 82 unrelated healthy control subjects without a family history of diabetes or PCOS (4 age- and weight-matched subgroups, i.e. Control(MothersPCOS), Control(FathersPCOS), Control(SistersPCOS), and Control(BrothersPCOS)) were studied. Glucose and insulin (at baseline and during a 75-g, 2-h oral glucose tolerance test) were measured. IR was assessed by fasting insulin (FI), fasting glucose to insulin ratio (FGI), homeostatic model assessment (HOMA IR), and area under the curve for insulin during the oral glucose tolerance test (AUC(insulin)) in NGT Mothers(PCOS), Fathers(PCOS), Sisters(PCOS), Brothers(PCOS), and matched control subgroups. Including the prestudy-diagnosed 3 mothers and 2 fathers with diabetes, diabetes and impaired glucose tolerance (IGT) were noted in 16% and 30% of Mothers(PCOS) and 27% and 31% of Fathers(PCOS), respectively. There was no diabetes in Sisters(PCOS) and Brothers(PCOS). IGT was found in 5% of Sisters(PCOS). Impaired fasting glucose was found in 3% of Mothers(PCOS) and 4% of Brothers(PCOS). The analysis of NGT family members showed that Mothers(PCOS) had higher FI (P < 0.05), HOMA IR (P < 0.05), and AUC(insulin) (P < 0.01) and lower FGI (P < 0.05) than Control(MothersPCOS), whereas all IR parameters were comparable between Fathers(PCOS) and their matched control subgroup. Sisters(PCOS) had higher FI (P < 0.05), HOMA IR (P < 0.01), and AUC(insulin) (P < 0.05) and lower FGI (P < 0.01), and Brothers(PCOS) had higher AUC(insulin) (P < 0.01) than their matched control subgroups, respectively. Mothers(PCOS) had higher testosterone levels than Control(MothersPCOS) (P < 0.01 and P < 0.05 for pre- and postmenopausal women, respectively). Sisters(PCOS) had higher LH (P < 0.01), testosterone (P < 0.001), androstenedione (P < 0.01), and dehydroepiandrosterone sulfate (P < 0.05) levels than Control(SistersPCOS). There was no difference in gonadotropin and androgen levels in Fathers(PCOS) compared with Control(FathersPCOS) or in Brothers(PCOS) compared with Control(BrothersPCOS). Our results suggest that 1) first degree relatives of patients with PCOS may be at high risk for diabetes and glucose intolerance; 2) NGT female family members have insulin resistance; and 3) mothers and sisters of PCOS patients have higher androgen levels than control subjects. We propose that the high risks of these impairments warrant screening in first degree relatives of patients with PCOS. PMID- 12727951 TI - Serum ghrelin levels in acromegaly: effects of surgical and long-acting octreotide therapy. AB - The orexigenic peptide, ghrelin, is regulated by acute and chronic nutritional state. Although exogenously administered ghrelin stimulates pituitary GH secretion, little is known about the role of ghrelin in endogenous GH secretion or how high GH and IGF-I levels in acromegaly could affect ghrelin secretion and vice versa. Therefore, we evaluated fasting and post oral glucose tolerance test serum ghrelin levels in 19 patients with active acromegaly at baseline and after either surgery in 9 of these or administration of long-acting octreotide (Sandostatin LAR) in the other 10 patients. After surgical cure, fasting ghrelin rose from 312 +/- 56 pg/ml to 548 +/- 97 pg/ml (P = 0.013). Fasting serum ghrelin levels were higher in all patients after surgery and ranged between 112% and 349% of presurgery levels. Ghrelin levels fell significantly during long-acting octreotide therapy from 447 +/- 34 pg/ml to 206 +/- 15 pg/ml (P < 0.0001); ghrelin levels on octreotide ranged between 26% and 70% of baseline levels. Serum ghrelin levels were suppressed significantly during the oral glucose tolerance test in both groups. Pretherapy ghrelin levels correlated negatively with serum insulin levels (r = -0.494; P = 0.03) and insulin resistance as estimated by the homeostasis model assessment score (r = -0.573; P = 0.01). In patients without diabetes mellitus, serum insulin levels in the surgical group were 19.7 +/- 5.4 microU/ml before surgery and fell to 9.7 +/- 0.93 microU/ml after surgery (P = 0.05); levels in the octreotide group were 13.9 +/- 2.8 microU/ml before and fell to 11.2 +/- 2.8 microU/ml on octreotide (P = 0.03). Pretherapy ghrelin levels did not correlate with weight or body mass index, but after therapy in the surgery group ghrelin correlated negatively with weight (r = -0.823, P = 0.012) as has been demonstrated by others in healthy subjects. Ghrelin secretion is dysregulated in active acromegaly; lowered serum levels of ghrelin in active acromegaly rise along with the postsurgery normalization of GH and IGF-I and improved insulin resistance. In contrast to surgical therapy, long-acting octreotide therapy persistently suppressed serum ghrelin levels. It remains to be determined whether altered circulating ghrelin concentrations could impact on body composition changes in acromegaly. PMID- 12727952 TI - Free cortisol index is better than serum total cortisol in determining hypothalamic-pituitary-adrenal status in patients undergoing surgery. AB - Serum total cortisol has traditionally been used for the interpretation of tests of the hypothalamic-pituitary-adrenal axis. Approximately 80% of total cortisol is bound to cortisol-binding globulin (CBG), and variation in CBG significantly affects serum total cortisol levels. Reliable assessment of hypothalamic pituitary-adrenal axis reserve is difficult in severely ill patients, because CBG falls substantially during the acute phase response. The free cortisol index (FCI), defined as the ratio of total cortisol/CBG, correlates well with serum free cortisol. We evaluated the FCI in the context of severe stress and the acute phase response by measuring total cortisol and CBG pre- and postoperatively in 31 patients undergoing major elective surgery. Serum total cortisol increased by 55% from 453 +/- 35.2 (mean +/- SEM) nmol/liter (range, 88-882) to 700 +/- 47.2 (range, 294-1631) nmol/liter. Serum CBG decreased by 30% from 45 +/- 1.7 (range, 26.6-64.1) to 31.4 +/- 1.62 (range, 16.1-51.9) mg/liter, but FCI increased by 130% from 10 +/- 0.8 (range, 2-18) to 23 +/- 1.7 (range, 13-58) nmol/mg. In seven patients (23%), postoperative serum total cortisol was less than 500 nmol/liter, but their postoperative CBG levels were significantly lower than levels in the rest of the group (P < 0.01). However, there was no difference in the FCI between this subgroup and the rest of the group. This study demonstrates the importance of CBG measurement and the calculation of FCI for the interpretation of serum total cortisol in situations where CBG changes significantly. PMID- 12727953 TI - Prostate volume and growth in testosterone-substituted hypogonadal men are dependent on the CAG repeat polymorphism of the androgen receptor gene: a longitudinal pharmacogenetic study. AB - Testosterone (T) substitution in hypogonadal men results in growth of the prostate gland. T effects are mediated via the androgen receptor (AR). The length of the (CAG)n polymorphism of the AR gene is negatively associated with transcriptional activity and might account for variations in prostate growth during substitution therapy. In 131 hypogonadal men aged 18-69 yr, we assessed prostate volume longitudinally by transrectal ultrasonography and determined AR (CAG)n, sex hormone levels, and anthropometric measures. Sixty-nine men with primary and 62 with secondary hypogonadism began substitution therapy with im injections of T enanthate (n = 81), transdermal T preparations (n = 19), sc injections of human chorionic gonadotropin (n = 17), or oral T undecanoate (n = 14) for 2.4 +/- 0.8 yr. Average prostate size increased from 15.8 +/- 6.1 ml to 23.0 +/- 6.8 ml. ANOVA including covariates revealed initial prostate size to be dependent on age (P < 0.001) and baseline T levels (P = 0.01) but not on number of (CAG)n (ranging from 13-30; mean, 21.4 +/- 3.5). Prostate growth per year and absolute prostate size under substituted T levels (6.1 +/- 3.3 to 21.6 +/- 10.3 nmol/liter) were strongly dependent on (CAG)n, with lower treatment effects in longer repeats (both P < 0.001). Other significant predictors were initial prostate size (negative for growth rate and positive for absolute size) and age (positive for both growth rate and absolute size). The odds ratio for men with (CAG)n less than 20, compared with those with (CAG)n of 20 or more to develop a prostate size of at least 30 ml under T substitution, was 8.7 (95% confidence interval, 3.1-24.3; P < 0.001). This observation was strongly age dependent with a more pronounced odds ratio in men older than 40 yr. This first pharmacogenetic study on androgen substitution in hypogonadal men demonstrates a marked influence of the AR gene (CAG)n polymorphism on prostate growth. PMID- 12727954 TI - Insulin-mediated hepatic glucose uptake is impaired in type 2 diabetes: evidence for a relationship with glycemic control. AB - Impaired hepatic glucose uptake (HGU) has been implicated in the development of hyperglycemia in type 2 diabetes; the relative impact of plasma glucose and insulin levels on this process remains controversial. We compared the effects of euglycemic hyperinsulinemia on HGU, skeletal muscle glucose uptake, and hepatic influx rate-constant (H-Ki) in 38 diet-treated diabetic patients and 22 nondiabetic controls, using positron emission tomography with (18)F fluorodeoxyglucose and the insulin clamp technique. Control subjects were divided into two subgroups: one including older, heavier, insulin-resistant controls (whole-body glucose uptake, M = 21.4 +/- 5.4 micromol x min(-1) x kg(-1)) to match characteristics of diabetic patients (M = 20.4 +/- 9.9); the other including younger, leaner, insulin-sensitive controls (M = 48.2 +/- 9.9, P < 0.01). Skeletal muscle glucose uptake showed a similar group distribution as the M value. Insulin clearance rates were lower, whereas glycosylated hemoglobin and clamp plasma insulin levels were higher in diabetic patients than in controls. HGU and H-Ki were similar in the two nondiabetic subgroups and lower in diabetic patients than in controls (1.9 +/- 0.5 vs. 2.3 +/- 0.7 micromol x min(-1) x 100 ml(-1), and 0.37 +/- 0.09 vs. 0.44 +/- 0.14 ml x min(-1) x 100 ml(-1), P < or = 0.01). In the whole dataset, H-Ki was inversely related to fasting plasma glucose (correlation coefficient = -0.40, P = 0.0018). In diabetic subjects, H-Ki was reciprocally related to glycosylated hemoglobin (correlation coefficient = -0.36, P = 0.029). We conclude that insulin-mediated HGU is impaired, in type 2 diabetes, in some proportion to the degree of glycemic control. PMID- 12727955 TI - Five years of growth hormone replacement therapy in adults: age- and gender related changes in isometric and isokinetic muscle strength. AB - GH replacement therapy in adults with adult-onset GH deficiency (GHD) has been shown to increase isometric and isokinetic muscle strength in a few trials with limited numbers of patients. In this single center, prospective, open-label study, the effects of 5-yr GH replacement therapy on muscle function were determined in 109 consecutive adults (61 men and 48 women) with adult-onset GHD. The mean initial GH dose was 0.88 mg/d. The dose was gradually lowered, and after 5 yr the mean dose was 0.46 mg/d. The mean IGF-I SD score increased from -1.54 at baseline to 1.53 at study end. A sustained increase in lean body mass and decrease in body fat was observed. The GH treatment induced persistent increases in isometric knee flexor strength, concentric knee flexor strength at an angular velocity of 60 degrees/sec, and right-hand peak grip strength. After correction for age and gender using observed/predicted value ratios, a sustained increase was also observed in isometric (60 degrees) and concentric (180 degrees/sec) knee extensor strength, average right-hand grip strength for 10 sec, and left-hand grip strength. At study end, knee flexor and extensor strength was 96-104% of predicted and hand grip strength was 84-90% of predicted values. The local muscle endurance was transiently decreased after correction for age and gender. No gender difference was found in the treatment responses in muscle strength. However, muscle strength (also after correction for age and gender) was lower in women than men throughout the study period. In conclusion, GH replacement therapy in adults with adult-onset GHD normalized isometric and isokinetic knee flexor and extensor strength. Hand grip strength increased but was not fully normalized. PMID- 12727956 TI - Rationale for central and bilateral lymph node dissection in sporadic and hereditary medullary thyroid cancer. AB - A retrospective study was performed on 101 consecutive medullary thyroid cancer (MTC) patients who underwent at Institut Gustave-Roussy (IGR) total thyroidectomy with central and bilateral lymph node dissection. At histology, lymph node metastases were found in 55% of patients. In sporadic MTC, lymph node metastases were observed in the central compartment in 50% of patients, in the ipsilateral jugulocarotid chain in 57%, and in the contralateral jugulocarotid chain in 28%. In hereditary MTC, lymph node metastases were identified in the central compartment in 45% of patients, in the ipsilateral jugulocarotid chain in 36%, and in the contralateral jugulocarotid chain in 19%. Contralateral lymph nodes were found in 37% of metastatic patients with an unilateral tumoral involvement of the thyroid gland. A strong association was observed between tumor size and lymph node involvement for both hereditary and sporadic MTC (P < 0.02). Permanent hypoparathyroidism occurred in 4% of patients and laryngeal nerve palsy in 5%. An undetectable calcitonin level was obtained after surgery in 61% of patients, in 95% of patients without lymph node metastases, and in 32% of patients with lymph node metastases. Among patients with lymph node involvement, undetectable calcitonin level was obtained in 57% of patients with less than or with 10 lymph node metastases and in 4% of patients with more than 10 (P < 0.01). IN CONCLUSION: 1) lymph node metastases occur early in the course of MTC; 2) the pattern of lymph node metastatic distribution in neck areas varied between patients and was not related to the thyroid tumor size; 3) contralateral lymph node metastases were observed even in patients with small thyroid tumor; 4) total thyroidectomy with central and complete bilateral neck dissection should be performed routinely in all patients with sporadic and hereditary MTC, even in those with small thyroid tumors-a contralateral neck dissection may be avoided only in sporadic MTC patients with unilateral involvement of the thyroid gland in the absence of central and ipsilateral neck involvement; and 5) the number of lymph node metastases was predictive of biological cure after surgery. PMID- 12727957 TI - Nephrolithiasis in Cushing's disease: prevalence, etiopathogenesis, and modification after disease cure. AB - The pathogenesis of nephrolithiasis in Cushing's syndrome is still not completely clarified. The current study aimed at investigating prevalence of nephrolithiasis and role of different lithogenic factors in Cushing's disease (CD). Forty-six CD patients (24 with active and 22 with cured disease) and 46 sex- and age-matched controls entered the study. Body mass index, blood pressure, fasting glucose and insulin, serum and urinary creatinine, urea, uric acid, electrolytes, and cystine, urinary volume, pH, oxalate, and citrate levels, and renal ultrasonography (US) were performed in all patients and controls. Nephrolithiasis was found in 50% of active patients, 27.3% of cured patients, and 6.5% of controls (P < 0.001). Compared with controls, patients with active disease had a significantly increased prevalence of obesity, arterial hypertension, diabetes mellitus, hypercalciuria, hypocitraturia, and hyperuricosuria, significantly higher levels of serum and urinary cystine, urinary creatinine, urea, uric acid, potassium, calcium, phosphorus, and oxalate, significantly lower levels of urinary citrate levels. Compared with controls, patients cured from CD had a significantly increased prevalence of obesity, systemic arterial hypertension, and diabetes mellitus, whereas urinary citrate was significantly decreased. At multivariate analysis, a significantly increased risk to develop kidney stones was independently associated with urinary excretion of uric acid (odds ratio = 1.6, confidence interval = 1.0-2.5) and systemic arterial blood pressure (odds ratio = 2.6, confidence interval = 1.1-6.6). In conclusion, patients with active CD have an increased prevalence of nephrolithiasis compared with general population, which decreases but not disappears in patients successfully cured from the disease. This complication is likely caused by the synergic effect of different hypercortisolism-dependent metabolic and hemodynamic abnormalities, among which systemic arterial hypertension and excessive urinary uric acid excretion seem to play a pivotal role. PMID- 12727958 TI - Torus palatinus: a new anatomical correlation with bone density in postmenopausal women. AB - The observation that subjects who have a striking oral exostosis, called torus palatinus, also tended to have normal or high bone densities prompted us to examine an unselected population referred for bone density assessment for a possible correlation with torus palatinus. Subjects referred from community physicians had a visual examination of the open mouth to estimate the size of any torus palatinus (0 for none/trace to 5 for very large) before undergoing a bone density measurement by dual energy x-ray absortiometry. Bone density T- and z scores were correlated with the size of each subject's torus palatinus. Torus size groups were also correlated with other variables affecting bone density. About 20% of 370 postmenopausal female subjects, >90% Caucasian, had a moderate to large torus palatinus. Regression correlations for torus size were modest, but significantly related to T- and z-scores of lumbar vertebrae and left hip (P < 0.01 for each). Differences due to medication, body mass index, smoking, parity, and several other factors that affect bone density did not diminish the relation to torus size. This study shows a small, but significant, positive relation for postmenopausal, Caucasian women between bone mineral density and torus size after controlling for several variables known to affect bone density were examined. Torus prominence, in association with other factors, can be considered in decisions for testing bone density in otherwise normal postmenopausal women. PMID- 12727959 TI - Impaired nighttime sleep in healthy old versus young adults is associated with elevated plasma interleukin-6 and cortisol levels: physiologic and therapeutic implications. AB - IL-6 and TNF alpha secretion is increased by sleep loss or restriction. IL-6 secretion progressively increases with age, yet its association with decreased quality and quantity of sleep in old adults is unknown. This study examined the alteration of 24-h secretory pattern of IL-6, TNF alpha, and cortisol in 15 young and 13 old normal sleepers who were recorded in the sleep laboratory for four consecutive nights. Serial 24-h plasma measures of IL-6, TNF alpha, and cortisol were obtained during the fourth day, and daytime sleepiness was assessed with the multiple sleep latency test. Old adults, compared with young subjects, slept poorly at night (wake time and percentage stage 1 sleep were increased, whereas their percentage slow wave sleep and percentage sleep time were decreased, P < 0.05). Accordingly, their daytime sleep latency was longer than in young adults (P < 0.05). The mean 24-h IL-6 and cortisol levels were significantly higher in old than young adults (P < 0.05). In both groups, IL-6 and cortisol plasma concentrations were positively associated with total wake time, with a stronger association of IL-6 and cortisol with total wake time in the older individuals (P < 0.05); their combined effect was additive. IL-6 had a negative association with rapid eye movement (REM) sleep only in the young (P < 0.05), but cortisol was associated negatively with REM sleep both in the young and old, with a stronger effect in the young. We conclude that in healthy adults, age-related alterations in nocturnal wake time and daytime sleepiness are associated with elevations of both plasma IL-6 and cortisol concentrations, but REM sleep decline with age is primarily associated with cortisol increases. PMID- 12727960 TI - Ultrasound evaluation of carotid artery in primary hyperparathyroidism. AB - Patients with primary hyperparathyroidism (PHPT) have an increased cardiovascular morbidity and mortality. Elevated serum calcium and/or PTH may directly contribute to vascular tissue damage, but the role of classic factors for atherosclerosis has not fully been evaluated in this disease. The aim of our study was to dissect the potential effect of hypercalcemia and/or high PTH from that of major cardiovascular risk factors (i.e. diabetes mellitus, hyperlipidemia, hypertension, obesity, smoking habit) on the carotid artery structure of patients with PHPT. Twenty-six consecutive patients with PHPT [subdivided into two groups according to the absence (n = 10) or the presence (n = 16) of one or more risk factors] and 15 normocalcemic healthy subjects as controls were studied. At ultrasonography, a significant increase (P < 0.001) of carotid mean and maximum intima-media thickness, as well as a significant reduction of lumen diameter (P < 0.05) were found in the PHPT group with risk factors, compared with the other two groups. This suggests that hypercalcemia and/or PTH elevation per se are not determinant of carotid atherosclerosis in PHPT, and that increased cardiovascular mortality and morbility in this disease is attributable to the combined presence of classic cardiovascular risk factors. PMID- 12727961 TI - Clinical features and outcome of subacute thyroiditis in an incidence cohort: Olmsted County, Minnesota, study. AB - Subacute thyroiditis (SAT), or granulomatous thyroiditis, is an inflammatory thyroid condition associated with pain and systemic symptoms. Few community studies are available. We studied the 160 patients with SAT in Olmsted County, Minnesota, seen between January 1, 1960, and December 30, 1997. Subjects were identified through the medical diagnostic index of the Rochester Epidemiology Project. The overall age- and sex-adjusted incidence from 1960 through 1997 was 4.9 cases per 100,000/yr. In the most recent 28-yr period (1970-1997), 94 patients were identified. In this group, pain was the presenting symptom in 96%. SAT recurred in 4% of the patients 6-21 yr after the initial episode. Corticosteroid therapy was given to 36%. Early-onset hypothyroidism occurred both in patients receiving corticosteroid therapy (29%) and in those not receiving corticosteroid therapy (37%). At latest follow-up, significantly more patients who had received corticosteroid therapy had a diagnosis of hypothyroidism than the group without corticosteroid therapy (25% vs. 10%, P < 0.05; overall rate of hypothyroidism, 15%). Early transient hypothyroidism is common in SAT. Permanent hypothyroidism is less common, and only 15% of the patients are receiving T(4) therapy after 28 yr of follow-up. Symptomatic relief is achieved with corticosteroid therapy, but such therapy does not prevent early- and late-onset thyroid dysfunction. PMID- 12727962 TI - Ten years on: Safety of short synacthen tests in assessing adrenocorticotropin deficiency in clinical practice. AB - Since 1988, when a retrospective study of patients attending this unit was published, we have advocated the use of the short synacthen test (SST) as the primary screening investigation to detect ACTH deficiency. However, others have published comparisons of SST and insulin tolerance tests that suggest a significant false negative rate with SST, leading to concern that some patients who pass the SST are in danger from the clinical consequences of ACTH deficiency. To address this, we audited biochemical results and clinical outcome in 63 patients who did not have ACTH deficiency detected (i.e. who passed the test) by SST after pituitary surgery. Twelve of the 63 patients who passed a SST after pituitary surgery became ACTH-deficient later as diagnosed by SST: 4 within the first year, 2 of whom had received postoperative radiotherapy (3 had symptoms of tiredness and 1 was admitted to the hospital with a viral infection); 8 in yr 3 5, 7 of whom had received postoperative radiotherapy (all had either no symptoms or symptoms of tiredness alone). Thus, the predictive value of the SST in excluding ACTH deficiency is approximately 97% (2 of 63 patients who initially passed the SST were found to be ACTH-deficient within 12 months without having received postoperative radiotherapy). Only 1 patient was ill enough to require hospital admission. Setting the risk of false negatives with SST against the morbidity and manpower implications associated with insulin tolerance tests, SST remains the primary screening test for ACTH deficiency in our practice. However, a high index of clinical suspicion to detect false negative results must be maintained. PMID- 12727963 TI - Low-dose growth hormone inhibits 11 beta-hydroxysteroid dehydrogenase type 1 but has no effect upon fat mass in patients with simple obesity. AB - GH has potent effects on adipocyte biology, stimulating lipolysis but also promoting preadipocyte proliferation. In addition, GH, acting through IGF-I, inhibits 11 beta-hydroxysteroid dehydrogenase type 1 (11 beta-HSD1), which converts the inactive glucocorticoid, cortisone (E), to active cortisol (F) in adipose tissue. Although F is an essential requirement for adipocyte differentiation, it also inhibits preadipocyte proliferation. We hypothesized that inhibition of 11 beta-HSD1 activity in adipose tissue by GH may alter fat tissue mass through changes in local F concentrations. We conducted a randomized, double-blind, placebo-controlled study using low-dose GH (Genotropin 0.4 mg/d) for 8 months in 24 patients with obesity. Although GH treatment significantly raised IGF-I, we were unable to demonstrate significant differences in body composition or metabolic profiles between GH- and placebo-treated groups. In addition, there was no alteration in total fat mass over time in the GH-treated group [total fat mass 41.0 +/- 3.0 vs. 41.3 +/- 3.4 kg (8 months), mean +/- SE, P = ns]. However, in comparison with baseline values, systolic blood pressure increased (119 +/- 3 vs. 130 +/- 4 mm Hg, P < 0.05 vs. baseline) and serum F/E ratio decreased (6.1 +/- 0.5 vs. 3.9 +/- 0.5, P < 0.05 vs. baseline) in the GH treated group only. Furthermore, although the urinary tetrahydrometabolites of F/E ratio fell in the GH-treated group, it rose in the placebo group (mean ratio change, -0.13 +/- 0.05 vs. +0.09 +/- 0.09, GH vs. placebo, P = 0.07). Treatment with low-dose GH in obesity fails to alter fat mass despite a significant elevation in IGF-I and a shift in the global set point of E to F conversion consistent with inhibition of 11 beta-HSD1. PMID- 12727964 TI - Within-subject between-cycle variability of histological dating, alpha v beta 3 integrin expression, and pinopod formation in the human endometrium. AB - alpha v beta 3 integrin expression and pinopod formation have been proposed as a means of distinguishing receptive endometrium from nonreceptive in clinical practice. However, one of the most intriguing facts in infertility is whether one cyclic event may be representative of all patients' cycles, and no study has evaluated the cycle-to-cycle variability of the expression of any of these new proposed markers in human endometrium. We investigated histological dating, alpha v beta 3 integrin expression, and pinopod formation in 45 endometrial biopsy specimens obtained in 15 primary infertility patients. All patients underwent three endometrial biopsies in consecutive spontaneous cycles, regardless of the previous histological findings. All endometrial samples were obtained on postovulatory d 7 as determined by ultrasonography. Agreement between the repeated observations (first vs. second, and first vs. third biopsies) with regard to histological dating and the presence or absence of alpha v beta 3 integrin and pinopods in the endometrium was analyzed using the 2 x 2 frequency tables and Cohen's kappa coefficient. The kappa values ranged from a low of -0.39 (level of agreement, poor) for alpha v beta 3 integrin expression to a high of 0.32 (level of agreement, fair) for biopsy dating by anticipated window of implantation. Overall, these results demonstrate that all endometrial variables investigated had poor reproducibility and high variability cycle to cycle. PMID- 12727965 TI - Relative effect of gonadotropin-releasing hormone (GnRH)-I and GnRH-II on gonadotropin release. AB - Two forms of GnRH (GnRH-I and GnRH-II) are expressed in the hypothalamus of humans and rhesus monkeys, but their relative abilities to stimulate LH and FSH release are unknown. Therefore, young (8-12 yr) and old (21-23 yr) female rhesus monkeys were treated i.v. with bolus injections of either GnRH-I or GnRH-II (dose range, 0.01-10 microg/kg body weight); serial blood samples were remotely collected through a vascular catheter for up to 2 h after injection. Overall, plasma LH concentrations were similarly elevated after treatment with GnRH-I and GnRH-II, and the responses were slightly greater in the younger animals. Although plasma FSH concentrations were unaffected by a single exposure to GnRH-I or GnRH II, they showed a similar significant increase after repeated exposures (every 2 h for 24 h). In a subsequent experiment, antide, a GnRH-I receptor antagonist, was administered (100 microg/kg body weight) together with a single injection of GnRH-I or GnRH-II (1 microg/kg body weight). As expected, GnRH-I-induced LH release was significantly attenuated by this combined treatment; moreover, GnRH II-induced LH release was completely blocked. Taken together, these data show that GnRH-II can potently stimulate gonadotropin release in vivo and that this action is likely mediated through the GnRH-I receptor. PMID- 12727966 TI - Endothelial function and menopause: effects of raloxifene administration. AB - Postmenopausal women have more severe endothelial dysfunction than premenopausal women. In the present study, we evaluated the possible beneficial effect of raloxifene administration, a selective estrogen receptor modulator, on endothelial regulation in postmenopausal women. In a double-blind, randomized vs. placebo trial, 60 healthy postmenopausal women were treated with raloxifene (60 mg/d) or placebo for 4 months to evaluate the effect of raloxifene treatment on endothelial function. Furthermore, in raloxifene-treated subjects (n = 30), the effect of raloxifene was also assessed during the intraarterial infusion of N(G) monomethyl-L-arginine (4 micromol/min). Raloxifene administration vs. placebo was associated with a decrease in plasma low-density lipoprotein cholesterol (P < 0.01), triglyceride (P < 0.05), thiobarbituric acid-reactive substance (P < 0.01), vascular cell adhesion molecule-1 (P < 0.05), intercellular adhesion molecule-1 (P < 0.001), and E-selectin (P < 0.001) levels and with an increase in plasma Trolox equivalent antioxidant capacity (P < 0.001) levels. Indeed, raloxifene treatment was also associated with a significant improvement in endothelial-dependent vasodilatation assessed by brachial reactivity technique. Raloxifene administration had no impact on endothelial-independent vasodilatation. Furthermore, intraarterial infusion of N(G)-monomethyl-L-arginine inhibited the significant effect of raloxifene on endothelium-mediated brachial arterial diameter and flow. In conclusion, our results demonstrate that raloxifene administration is associated with a positive modulation of endothelial dependent vasodilatation likely due to a reduction of risk factors for endothelial damage. PMID- 12727968 TI - Cushing's syndrome secondary to adrenocorticotropin-independent macronodular adrenocortical hyperplasia due to activating mutations of GNAS1 gene. AB - ACTH-independent macronodular adrenal hyperplasia (AIMAH) is an uncommon cause of Cushing's syndrome characterized by bilateral nodular adrenocortical hyperfunction in the presence of suppressed ACTH levels. We investigated whether activating mutations in the ACTH receptor (MC2-R) or G(s alpha) (GNAS1) genes might be involved in AIMAH genesis. Five women with Cushing's syndrome due to AIMAH, confirmed by histological studies, and no signs of McCune-Albright syndrome were selected for molecular analysis of these genes. The single exon of the MC2-R gene and exons 8 and 9 of the GNAS1 gene were amplified by PCR in genomic DNA from adrenal nodules and peripheral blood. Direct sequencing revealed only MC2-R wild-type sequences. GNAS1 PCR products at denaturing gradient gel electrophoresis revealed abnormal migration patterns in adrenal tissues of three patients. Automatic sequencing showed two different activating mutations at codon Arg(201) of GNAS1, a substitution by histidine in two cases and by serine in one case. In conclusion, we found two different gsp mutations in three patients with Cushing's syndrome due to AIMAH, and we speculate whether they belong to the spectrum of McCune-Albright syndrome or whether these are the first reported cases of AIMAH due to gsp mutations. PMID- 12727967 TI - Frequency of antineutrophil cytoplasmic antibody in Graves' disease patients treated with methimazole. AB - Retrospective studies have shown antineutrophil cytoplasmic antibody (ANCA) positivity in patients treated for Graves' hyperthyroidism; ANCA has been attributed to either antithyroid drugs or to the disease itself. The aim of this study was to determine ANCA in Graves' disease patients at diagnosis and after treatment with methimazole and to evaluate the relationship between ANCA and hyperthyroidism evolution. Thirty patients recently diagnosed with Graves' hyperthyroidism were prospectively studied. ANCA were determined by indirect immunofluorescence. ANCA autoantibodies against specific antigens (proteinase 3, myeloperoxidase, bactericidal/permeability-increasing protein (BPI), cathepsin, lysozyme, elastase, and lactoferrin) were detected by ELISA. The median observation period was 22 months. Kaplan-Meier analysis was performed to identify ANCA as an outcome variable. Twenty patients (67%) were ANCA positive before the onset of treatment, and four (19%) remained positive after 1 yr of antithyroid drug treatment. No differences were observed in any clinical or analytical features between patients with or without positive ANCA. Before treatment, BPI positive patients required radioiodine treatment or presented relapse more rapidly than BPI-negative patients (log-rank test P < 0.0002). Patients with Graves' hyperthyroidism show positive ANCA before medical treatment, which points to a relationship with the autoimmune disease itself. Our results suggest that BPI-positive patients tend to relapse with antithyroid medication. PMID- 12727969 TI - Endothelial function and coagulant factors in growth hormone-treated hypopituitary adults receiving desmopressin. AB - Although GH deficiency may underlie the increased cardiovascular risk in adult hypopituitarism, other coexisting hormonal deficiencies and/or unphysiological hormone replacement may contribute. L-Deamino-8-D-arginine (DDAVP), when administered parenterally, potentiates hemostasis by increasing plasma procoagulant factors. We investigated whether chronic intranasal DDAVP therapy influences clotting factors (plasma fibrinogen, factor VIII, and von Willebrand factor antigen) and endothelial function (flow-mediated dilation of the brachial artery) in 30 GH-treated hypopituitary subjects, including both DDAVP-treated subjects (group A) (mean age, 46 +/- 11 yr) and vasopressin-sufficient subjects (group B) (mean age, 47 +/- 16 yr). Fifteen healthy controls (group C) (mean age, 48 +/- 12 yr) were also studied. All hypopituitary patients were receiving stable GH replacement (median duration, 19 months). Comparing the three groups, concentrations of fibrinogen (mean +/- SD) (A, 3.3 +/- 1.0 g/liter vs. B, 3.5 +/- 0.9 vs. C, 2.6 +/- 0.8, P < 0.05), factor VIII (A, 130% +/- 30% vs. B, 128% +/- 30% vs. C, 104% +/- 35%, P < 0.05) and von Willebrand factor antigen (A, 124% +/- 35% vs. B, 134% +/- 45% vs. C, 93% +/- 36%, P < 0.05) were higher in hypopituitary subjects, compared with controls. However, there were no differences in clotting factors between groups A and B. Flow-mediated dilation did not differ significantly between the two hypopituitary groups (A, 5.9% +/- 2.0% vs. B, 4.7% +/- 1.6%) and was similar to that in the control group (C, 5.7% +/- 2.1%). In conclusion, although endothelium-dependent vasodilation is intact in GH-treated hypopituitary adults, elevated concentrations of hemostatic markers suggest the persistence of a prothrombotic tendency and endothelial dysfunction. Intranasal DDAVP does not appear to influence this proatherogenic profile in hypopituitary adults with vasopressin deficiency. PMID- 12727970 TI - Coordinate regulation of the production and signaling of retinoic acid by estrogen in the human endometrium. AB - To determine whether estrogen regulates retinoic acid (RA) production and signaling in the human endometrium as it does in the rodent uterus, we investigated the effects of estrogens on the expression of RA-metabolizing enzymes, retinoid receptors, and biomarker genes in the post- and premenopausal human endometrium. Real-time quantitative PCR revealed that retinaldehyde dehydrogenase (RALDH) 2, a critical enzyme in RA biosynthesis, was induced 4-fold by estrogen replacement therapy with either Premarin or a mixture of estrone and equilin sulfates for 3 months. Estrogen replacement therapy also increased the expression of the RA receptor RAR alpha 1.9-fold. In parallel, there was a marked increase in the expression of two RA-regulated genes, cellular retinoic acid binding protein II and tissue transglutaminase. In the premenopausal endometrium, the levels of RALDH1, RALDH2, RAR alpha, and cellular retinoic acid-binding protein II were increased in the estrogen-dominated proliferative phase, and the transcripts for the RA catabolic enzyme retinoic acid 4-hydroxylase (CYP26A1) and tissue transglutaminase were significantly increased in the secretory phase. Our results suggest that estrogen coordinately up-regulates RA production and signaling in the human endometrium. This coordinate mechanism may play a role in the antiproliferative effects that counterbalance the estrogen-induced endometrial proliferation. PMID- 12727971 TI - Neurokinin B is a paracrine vasodilator in the human fetal placental circulation. AB - Neurokinin (NK) B is a member of the tachykinin family of neurotransmitters, exerting hypotensive or hypertensive effects in the mammalian vasculature through synaptic release from peripheral neurons, according to either NK(1) and NK(2) or NK(3) receptor subtype expression, respectively. There is recent evidence that NKB is expressed by the syncytiotrophoblast of the human placenta, an organ that is not innervated. We hypothesized that NKB is a paracrine modulator of tone in the fetal placental circulation. We tested this hypothesis using the in vitro perfused human placental cotyledon. Our data show that NKB is a dilator of the fetal vasculature, causing a maximal 25.1 +/- 4.5% (mean +/- SEM; n = 5) decrease in fetal-side arterial hydrostatic pressure (5- microM NKB bolus; effective concentration in the circulation, 1.89 nM) after preconstriction with U-46619. RT PCR demonstrated the presence of mRNA for NK(1) and NK(2) tachykinin receptors in the placenta. Using selective receptor antagonists, we found that NKB-induced vasodilation is through the NK(1) receptor subtype. We found no evidence for the involvement of either nitric oxide or prostacyclin in this response. This study demonstrates a paracrine role for NKB in the regulation of fetal placental vascular tone. PMID- 12727972 TI - N-terminal proopiomelanocortin acts as a mitogen in adrenocortical tumor cells and decreases adrenal steroidogenesis. AB - There is evidence that proopiomelanocortin (POMC)-derived peptides other than ACTH are involved in pituitary-dependent adrenal growth. We have synthesized the human N-terminal POMC fragment 1-28-POMC with the disulfide bridges in the correct position between cysteine residues 2-24 and 8-20 and studied the activity of these peptides in adrenocortical tumor cells in vitro. 1-28-POMC stimulated cell proliferation in human NCI-h295 and mouse Y-1 adrenal cancer cell lines and also in primary cultures of bovine adrenocortical cells in a concentration dependent manner. 1-28-POMC led to rapid activation of the MAPKs extracellular signal-regulated kinases-1 and -2, but not c-Jun N-terminal kinase and p38, pathways. Steroid hormone production (cortisol, 17-hydroxyprogesterone, and dehydroepiandrosterone sulfate) in NCI-h295 cells was decreased by 1-28-POMC in a concentration-dependent fashion. However, protein levels of important regulators of steroidogenesis [steroidogenic factor-1, DAX-1 (dosage-sensitive sex reversal adrenal hypoplasia congenita critical region on the X-chromosome 1), steroidogenic acute regulatory protein, and cytochrome P450 side-chain cleavage enzyme] remained unaffected by 1-28-POMC treatment. Our results provide evidence that synthetic 1-28-POMC induces adrenal tumor cell proliferation, inhibits adrenal steroidogenesis, and mediates its action by signaling via the extracellular signal-regulated kinase pathway. The distinct roles of 1-28-POMC and ACTH in the regulation of adrenal growth and steroidogenesis suggest that the adrenal cortex is under the dual opposing control of fragments from the same mother peptide POMC. PMID- 12727973 TI - Ghrelin secretion in humans is sexually dimorphic, suppressed by somatostatin, and not affected by the ambient growth hormone levels. AB - We studied plasma ghrelin and GH concentrations over a 24-h period in young healthy men and women and in patients with acromegaly. Healthy subjects were restudied after administration of GH-lowering agents, octreotide or GHRH antagonist. Ghrelin concentrations in women studied during the late follicular stage of the cycle were about 3-fold higher than in men. Suppression of GH secretion by GHRH antagonist did not alter ghrelin concentration profiles. In the presence of high GH levels (acromegaly), ghrelin levels were similar to those found in healthy men. Administration of somatostatin analog octreotide suppressed both GH and ghrelin concentration profiles. We conclude that: 1) ghrelin secretion is sexually dimorphic in humans, with women in the late follicular stage having higher levels than men; 2) ghrelin secretion is suppressed by somatostatin; and 3) GH has no influence over ghrelin secretion. PMID- 12727974 TI - The use of androgen receptor amino/carboxyl-terminal interaction assays to investigate androgen receptor gene mutations in subjects with varying degrees of androgen insensitivity. AB - Five mutations in the ligand-binding domain (LBD) of the human androgen receptor (hAR) found in patients with varying degrees of androgen insensitivity syndrome (AIS) were investigated for their effects on receptor dynamics. These were Arg(871)Gly (mild), Ser(814)Asn (partial), Glu(772)Ala (partial), Val(866)Met (complete), and Arg(774)Cys (complete). Previous analysis showed that the mutant receptors exhibited near-normal kinetics, except Arg(774)Cys, which had severely reduced androgen binding, and Val(866)Met, which showed increased equilibrium dissociation constant (K(d)) and elevated dissociation rate (k) values. Ser(814)Asn exhibited ligand-selective k values, i.e. increased for dihydrotestosterone and mibolerone, but normal for methyltrenolene. Using mammalian two-hybrid assays, hAR amino/carboxyl (N/C)-terminal interactions of the mutant receptors were analyzed in the presence and absence of the hAR coactivator transcription intermediary factor 2 (TIF2). The mutations conferred decreased hAR N/C-terminal interaction, i.e. mild (approximately 1.5-fold), partial (2-fold), and complete (10-fold), that mirrored the degree of AIS. All mutant LBDs showed a 2- to 3-fold increase in N/C-terminal interactions when TIF2 was cotransfected, although of a magnitude still less than that of wild-type LBD with TIF2. The ligand-selective properties of the Ser(814)Asn mutant were also clearly reflected by the N/C-terminal interactions. Thus, measurement of N/C terminal interactions may assist in the molecular analysis of mutant hARs associated with AIS. PMID- 12727975 TI - Human myometrial quiescence and activation during gestation and parturition involve dramatic changes in expression and activity of particulate type II (RII alpha) protein kinase A holoenzyme. AB - There are substantial data indicating that components of the cAMP-signaling pathway are differentially expressed in the human myometrium during pregnancy. The effects of cAMP in most tissues and cell types are mainly modulated via protein kinase A, a heterotetrameric protein complex consisting of two regulatory (R) and two catalytic (C) subunits. In the studies presented here, we used specific antibodies in Western blotting/immunoprecipitation, RT-PCR, and functional protein kinase A (PKA) phosphorylation assays to determine the PKA holoenzymes that are expressed in the human myometrium throughout pregnancy and labor. We report that as early as the second trimester of pregnancy, there is a significant increase in expression of the regulatory RII alpha protein subunit of PKA in the myometrium. This increase in protein expression is also mirrored at the mRNA level, indicating transcriptional control throughout pregnancy, whereas during parturition both transcript and protein are significantly decreased. This increase in RII alpha protein also resulted in increased particulate PKA activity in the myometrium during gestation, which was subsequently decreased during labor. Two specific A kinase anchoring proteins, AKAP95 and AKAP79, which have high binding affinities for RII alpha subunits, were found to form complexes with myometrial RII alpha species employing immunoprecipitation assays, but their levels of expression remained uniform in all myometrial tissue samples investigated. Our findings indicate that increased particulate type II PKA activity occurs throughout pregnancy, therefore directing the cAMP quiescence signal to specific subcellular loci within myometrial smooth muscle cells including the contractile machinery at the cytoskeleton; this effect is then removed during parturition. PMID- 12727976 TI - Effects of growth hormone on pulmonary function, sleep quality, behavior, cognition, growth velocity, body composition, and resting energy expenditure in Prader-Willi syndrome. AB - The objective of this study was to investigate the effects of GH administration on pulmonary function, sleep, behavior, cognition, linear growth velocity, body composition, and resting energy expenditure (REE) in children with Prader-Willi syndrome. The study used a 12-month, balanced, randomized, double-blind, placebo controlled, cross-over experimental design. Twelve subjects were randomized to GH (0.043 mg/kg x d) or placebo intervention for 6 months and then crossed over to the alternate intervention for 6 months. Differences in outcome variables were determined by paired t tests. Peak flow rate, percentage vital capacity, and forced expiratory flow rate improved and number of hypopnea and apnea events and duration of apnea events trended toward improvement after GH intervention. The only difference in cognition or behavior was an increase in hyperactivity scale on the Behavior Assessment System for Children after GH intervention. Linear growth velocity, REE, and lean mass were higher (67%, 19%, and 7.6%, respectively), and fat mass and percentage body fat were lower (10.3% and 8.1%, respectively) after GH intervention. GH administration did not change mean fasting ghrelin concentration. GH intervention improved body composition and REE and may contribute to better sleep quality and pulmonary function. GH administration did not impact fasting ghrelin concentration. PMID- 12727977 TI - Structure and function of disease-causing missense mutations in the PHEX gene. AB - The PHEX gene that is mutated in patients with X-linked hypophosphatemia (XLH) encodes a protein homologous to the M13 family of zinc metallopeptidases. The present study was undertaken to assess the impact of nine PHEX missense mutations on cellular trafficking, endopeptidase activity, and protein conformation. Secreted forms of wild-type and mutant PHEX proteins were generated by PCR mutagenesis; these included C85R, D237G, Y317F, G579R, G579V, S711R, A720T, and F731Y identified in XLH patients, and E581V, which in neutral endopeptidase 24.11 abolishes catalytic activity but not plasma membrane localization. The wild-type and D237G, Y317F, E581V, and F731Y proteins were terminally glycosylated and secreted into the medium, whereas the C85R, G579R, G579V, S711R, and A720T proteins were trapped inside the transfected cells. Growing the cells at 26 C permitted the secretion of G579V, S711R, and A720T proteins, although the yield of rescued G579V was insufficient for further analysis. Endopeptidase activity of secreted and rescued PHEX proteins, assessed using a novel internally quenched fluorogenic peptide substrate, revealed that E581V and S711R are completely inactive; D237G and Y317F exhibit 50-60% of wild-type activity; and A720T and F731Y retain full catalytic activity. Conformational analysis by limited proteolysis demonstrated that F731Y is more sensitive to trypsin and D237G is more resistant to endoproteinase Glu-c than the wild-type protein. Thus, defects in protein trafficking, endopeptidase activity, and protein conformation account for loss of PHEX function in XLH patients harboring these missense mutations. PMID- 12727978 TI - A genome-wide scan in type 2 diabetes mellitus provides independent replication of a susceptibility locus on 18p11 and suggests the existence of novel Loci on 2q12 and 19q13. AB - A genome-wide scan was performed, using nonparametric linkage analyses, to find susceptibility loci for type 2 diabetes mellitus in the Dutch population. We studied 178 families from The Netherlands, who constituted 312 affected sibling pairs. The first stage of the genome scan consisted of 270 DNA markers, with an average intermarker spacing of 13 cM. Because obesity and type 2 diabetes mellitus are interrelated, the data set was stratified for the subphenotype body mass index, corrected for age and gender. This resulted in a suggestive maximum multipoint LOD score of 2.3 (single-point P value, 9.7 x 10(-4); genome-wide P value, 0.028) for the most obese 20% pedigrees of the data set, between marker loci D18S471 and D18S843. In the lowest 80% obese pedigrees, two interesting loci on chromosome 2 and 19 were found, with LOD scores of 1.5 and 1.3. We provide independent evidence that the chromosome 18p11 locus, reported earlier from a Finnish/Swedish population, is of definite interest for type 2 diabetes mellitus in connection with obesity. Subsequently, our results indicate that two novel loci may reside on chromosomes 2 and 19, with minor effects involved in the development of type 2 diabetes mellitus in the Dutch population. PMID- 12727979 TI - Decreased type I collagen expression in human uterine cervix during pregnancy. AB - To provide some insight into the mechanism of cervical ripening, the expression of type I collagen was investigated in human uterine cervical tissues obtained from the first (n = 4) and third (n = 3) trimesters of normal pregnancy. Indirect immunofluorescent staining was performed for type I collagen, and Northern blot analysis was done to assess expression of mRNA for the alpha1(I) chain. Collagens were also extracted from the human cervical tissues in the first and third trimesters of pregnancy. Immunohistochemical analysis revealed loose distribution of type I collagen in the uterine cervix of the first trimester compared with the third trimester of pregnancy. The relative levels of various collagens were evaluated by SDS-PAGE. The ratios of the intensity of the band of alpha1(I) to that of total collagen alpha1 chain in cervical tissues of the third trimester were significantly lower than those in cervical tissues of the first trimester of pregnancy (P < 0.05). In contrast, the ratios of the intensity of the band of alpha1(III) to that of total collagen alpha1 chain in cervical tissues of the third trimester were significantly higher than those in cervical tissues of the first trimester of pregnancy (P < 0.05). Northern blot analysis revealed that the cervical expression of mRNA for the alpha1(I) chain was significantly reduced in the third trimester compared with the first trimester of pregnancy (P < 0.01). These results suggest that type I collagen might play an important role in the maintenance of pregnancy and that decreased expression of this collagen could be associated with the process of uterine cervical ripening. PMID- 12727980 TI - Endometrial stromal cells undergoing decidualization down-regulate their properties to produce proinflammatory cytokines in response to interleukin-1 beta via reduced p38 mitogen-activated protein kinase phosphorylation. AB - The decidualized endometrium plays a role in regulating trophoblast invasion for successful implantation and maintenance of pregnancy. IL-1 beta, a proinflammatory cytokine, has been suggested to play a role in this process. Recently, several lines of evidence indicate the importance of p38 MAPK in various inflammatory responses. We investigated whether endometrial stromal cells (ESC) change their inflammatory responses to IL-1 beta as related to p38 MAPK phosphorylation during the process of decidualization. ESC were decidualized by the treatment with progesterone for 9 d, as determined as such by an increase in the production of prolactin and cAMP by the cells. Whereas IL-1 beta increased the production of IL-6, IL-8, and monocyte chemotactic protein-1, and expression of cyclooxygenase-2 mRNA in ESC cultured without treatment, the stimulatory effects of IL-1 beta were reduced in the decidualized cells. Treatment with SB202190, a p38 MAPK inhibitor, also reduced the stimulatory effects of IL-1 beta in nondecidualized ESC. P38 MAPK phosphorylation was increased by IL-1 beta in nondecidualized ESC, whereas the IL-1 beta-induced increase was suppressed in the decidualized cells. Treatment with 8-bromo-cAMP reduced IL-1 beta-induced phosphorylation of p38 MAPK in nondecidualized ESC. In contrast, treatment with H89, a protein kinase A inhibitor, reversed a reduction in IL-1 beta-induced p38 MAPK phosphorylation in the decidualized cells. In summary, decidualization seems to be a process during which endometrial cells diminish their response to IL-1 beta, a known key factor for implantation, leading to the down-regulation of inflammation-like events, which may be relevant to controlled trophoblast invasion. The altered property of decidualized cells is thought to be caused by attenuation of IL-1 beta-induced p38 MAPK phosphorylation in a way that involves the activation of the cAMP/protein kinase A pathway. PMID- 12727981 TI - Absence of exon 10 of the human luteinizing hormone (LH) receptor impairs LH, but not human chorionic gonadotropin action. AB - The LH receptor (LHR) mediates the actions of LH and human chorionic gonadotropin (hCG). In vivo data showed that deletion of exon 10 does not affect hCG action, whereas LH action is impaired. To investigate the role of exon 10 in LH/hCG action in vitro we created stable COS-7 cells expressing the LHR with (wt) or without (-ex10) exon 10. Binding experiments showed that the affinities of LH and hCG to the LHR wt and -ex10 were similar. Stimulation of wt with hCG or LH resulted in increased cAMP. cAMP production was significantly impaired in -ex10 stimulated with LH. This response was not altered by pertussis toxin, excluding that G(i) becomes activated in LHR -ex10. In desensitization experiments, intracellular cAMP of LHR wt and -ex10 declined to approximately 30%. No difference in intracellular cAMP was detected between LHR wt or -ex10 after recovery and restimulation with hCG or LH. These experiments show that impaired cAMP production of LHR -ex10 stimulated with LH is not due to anomalous receptor coupling or desensitization. We conclude that although exon 10 of the LHR plays no role in ligand binding, it is important for receptor activation by LH by a mechanism probably involving extracellular conformational changes. PMID- 12727982 TI - Plasma levels of parathyroid hormone (1-84) whole molecule and parathyroid hormone (7-84)-like fragments in pseudohypoparathyroidism type I. AB - PTH (7-84) has antagonistic effects on the calcemic and phosphaturic actions of PTH (1-84) whole molecule (bioPTH). Human plasma contains bioPTH and PTH (7-84) like fragments. Using bioPTH-specific and nonspecific assays, we found that the patients with pseudohypoparathyroidism (PHP) type I with PTH-resistant hypocalcemia and hyperphosphatemia had the increased plasma levels of bioPTH and PTH (7-84)-like fragments than normal subjects (26.8 +/- 13.2 vs. 2.37 +/- 0.75 pmol/liter, P < 0.01 and 16.2 +/- 8.8 vs. 0.82 +/- 0.47 pmol/liter, P < 0.01, respectively). Calcitriol treatment increased phosphaturic response to PTH (1-34) (P < 0.05), and there was a negative correlation between phosphaturic response and the PTH levels (P < 0.05). These results suggested that the increased bioPTH and PTH (7-84)-like fragment levels may be related to the impaired phosphaturic response to PTH (1-34) in PHP type I. We also examined bioPTH-calcium dynamics in PHP type Ib patients and found that set-point calcium was 0.928 +/- 0.045 mmol/liter and the baseline to maximal ratio of bioPTH was 0.96 +/- 0.04. Calcitriol treatment increased set-point calcium to 1.129 +/- 0.028 mmol/liter (P < 0.01) and suppressed baseline to maximal ratio of bioPTH to 0.35 +/- 0.21 (P < 0.01). These bio-PTH calcium dynamics studies revealed the maximally stimulated baseline PTH secretion in PHP type Ib and demonstrated the effects of calcitriol on PTH-calcium curve shift and the degree of relative stimulation of baseline secretion. PMID- 12727983 TI - Mechanism of action in dogs of slow-acting insulin analog O346. AB - We compared metabolic effects as well as plasma and interstitial fluid kinetics of fatty acid-acylated insulin, Lys(B29)(N(epsilon)-omega-carboxynonadecanoyl) des(B30) human insulin (O346), with previously determined kinetics of native insulin and insulin detemir. Euglycemic clamps with iv injection of O346 (90 pmol/kg) or saline control were performed in 10 male mongrel dogs under inhalant anesthesia. The t(1/2) for the clearance of O346 from plasma was 375.7 +/- 26.7 min; the t(1/2) for the appearance of O346 in interstitial fluid was 137 +/- 20 min (mean +/- SEM). Glucose disposal with O346 injection was increased 4-fold (t = 480 min, 8.3 +/- 1.42 mg/min/kg) compared with preinjection (t = 0 min, 2.1 +/- 0.13 mg/min/kg; P < 0.05) or saline control (t = 480 min, 2.09 +/- 0.22 mg/min/kg; P < 0.05). O346 plasma elimination and transendothelial transport were 0.3% and 3.5% of regular insulin and 3% and 50% of insulin detemir, respectively. Combination of in vivo results and compartmental modeling suggests that the duration of action of O346 after iv injection is about 25-fold and 10-fold longer compared with regular human insulin and insulin detemir, respectively. This study demonstrates that O346 stimulates glucose disposal very slowly, but when injected iv, its effect may be maintained for as long as 48 h as estimated from simulation analysis. The data suggest that O346 bound to albumin in plasma acts as a storage compartment for O346 from which the analog is slowly released to insulin sensitive tissues. Reduced liver clearance of O346 is suggested to be the major mechanism for the protracted action. PMID- 12727984 TI - Platelet dysfunction in lean women with polycystic ovary syndrome and association with insulin sensitivity. AB - Platelet dysfunction and its association with insulin resistance and/or hyperandrogenemia were evaluated in 50 women with polycystic ovary syndrome (PCOS), 50 women with non-classic congenital adrenal hyperplasia (NC-CAH), and 30 women in the control group. Agonist-induced platelet aggregation was measured. Women with PCOS had significantly higher levels of platelet aggregations induced by ADP (77.4 +/- 3.3 vs. 67.3 +/- 2.8), collagen (79.7 +/- 1.8 vs. 69.1 +/- 3.9), and epinephrine (84.7 +/- 2.6 vs. 67.8 +/- 3.8), compared with controls. However platelet aggregations of women with NC-CAH because of ADP (68.2 +/- 4.22), collagen (69.5 +/- 5.4), or epinephrine (68.6 +/- 4.3) were similar to those in the control group. There were negative correlations between aggregations induced by agonists and the insulin sensitivity in women with PCOS. These correlations also appeared significant after androgen levels with covariance analysis were excluded. These covariance analyses were performed because serum androgen levels might affect platelet function. Any significant correlations were not found between androgen levels and agonist-induced platelet aggregation in women with NC CAH. We conclude that platelet dysfunction may be an important reason for the possible cardiovascular heart diseases in women with PCOS. PMID- 12727985 TI - Mechanisms behind lipolytic catecholamine resistance of subcutaneous fat cells in the polycystic ovarian syndrome. AB - Lipolytic catecholamine resistance in sc fat cells is observed in polycystic ovarian syndrome (PCOS). The mechanisms behind this lipolysis defect were explored in vitro; sc fat cells were obtained from 10 young, nonobese PCOS women and from 14 matched, healthy control women. Fasting plasma glycerol levels were reduced by one third in PCOS (P < 0.05). Adipocytes of PCOS women were about 25% larger than in the controls (P < 0.05) and had 40% reduced noradrenaline-induced lipolysis (P < 0.05), which could be attributed to a 10-fold decreased beta(2) adrenoceptor sensitivity (P < 0.05) and low ability of cAMP to activate the protein kinase A (PKA)/hormone-sensitive lipase (HSL) complex (P < 0.05). In PCOS, the adipocyte protein content of beta(2)-adrenoceptors, HSL, and the regulatory II beta-component of PKA were 70%, 55%, and 25% decreased, respectively (P < 0.001); but there was no change in the amount of the catalytic subunit of PKA or of beta(1)-adrenoceptors. Thus, lipolytic catecholamine resistance of sc adipocytes in PCOS is probably attributable to a combination of decreased amounts of beta(2)-adrenergic receptors, the regulatory II beta component of PKA, and HSL. This may cause low in vivo lipolytic activity and enlarged sc fat cell size and promote later development of obesity in PCOS. PMID- 12727986 TI - PDS is a new susceptibility gene to autoimmune thyroid diseases: association and linkage study. AB - Autoimmune thyroid disease (AITD), including Graves' disease (GD), Hashimoto thyroiditis (HT), and primary idiopathic myxedema, is caused by multiple genetic and environmental factors. Genes involved in immune response and/or thyroid physiology appear to influence susceptibility to disease. The PDS gene (7q31), responsible for Pendred syndrome (congenital sensorineural hearing loss and goiter), encodes a transmembrane protein known as pendrin. Pendrin is an apical porter of iodide in the thyroid. To evaluate the contribution of PDS gene in the genetic susceptibility of AITD, we examined four microsatellite markers in the gene region. Two hundred thirty-three unrelated patients (GD,141; HT, 54; primary idiopathic myxedema, 38), 15 multiplex AITD families (104 individuals/46 patients) and 154 normal controls were genotyped. Analysis of case-control data showed a significant association of D7S496 and D7S2459 with GD (P = 10(-3)) and HT (P = 1.07 10(-24)), respectively. The family-based association test showed significant association and linkage between AITDs and alleles 121 bp of D7S496 and 173 bp of D7S501. Results obtained by transmission disequilibrium test are in good agreement with those obtained by the family-based association test. Indeed, evidence for linkage and association of allele 121 bp of D7S496 with AITD was confirmed (P = 0.0114). Multipoint nonparametric linkage analysis using MERLIN showed intriguing evidence for linkage with marker D7S496 in families with only GD patients [Z = 2.12, LOD = 0.81, P = 0.026]. Single-point and multipoint parametric LOD score linkage analysis was also performed. Again, the highest multipoint parametric LOD score was found for marker D7S496 (LOD = 1.23; P = 0.0086) in families segregating for GD under a dominant model. This work suggests that the PDS gene should be considered a new susceptibility gene to AITDs with varying contributions in each pathology. PMID- 12727987 TI - Differential regulation of proteasome-dependent estrogen receptor alpha and beta turnover in cultured human uterine artery endothelial cells. AB - Estrogen-induced loss of estrogen receptor (ER) alpha expression limits estrogen responsiveness in many target cells. However, whether such a mechanism contributes to changes in vascular endothelial ER alpha and/or ER beta levels is unclear. Using RT-PCR assays, we did not find any regulation of ER alpha or ER beta mRNA expression in human uterine artery endothelial cell (HUAEC) nuclear extracts on stimulation with 17 beta-estradiol for 1 or 2 h. By contrast, Western analysis on HUAEC extracts revealed that 17 beta-estradiol was capable of down regulating both ER alpha and ER beta protein starting 1 h after treatment, an effect that can be blocked by pretreatment with tamoxifen as well as with the proteasome inhibitor lactacystin. The proteolysis inhibitors insulin, cycloheximide, and puromycin impede ER alpha, but not ER beta, turnover. Ubiquitin, but not its competitive inhibitor methyl-ubiquitin, induces rapid turnover of both ERs in a cell-free system of MCF-7 and HUAEC extracts. We, thus, propose the existence of estrogen-induced ER degradation that serves to control physiological responses in an estrogen target tissue, i.e. human vascular endothelium, by down- regulating ER alpha as well as ER beta through different proteasomal uptake mechanisms. PMID- 12727988 TI - Down-regulation of steroidogenic response to gonadotropins in human and rat preovulatory granulosa cells involves mitogen-activated protein kinase activation and modulation of DAX-1 and steroidogenic factor-1. AB - Gonadotropins were recently demonstrated to be able to activate the MAPK cascade, but the physiological significance of this activation is still obscure. In the present work we demonstrate that highly luteinized human granulosa cells obtained from in vitro fertilization patients respond to human LH as well as to forskolin in phosphorylation of extracellular-signal regulated kinases 1 and 2 (ERK1 and 2). Moreover, the potent MAPK inhibitors, PD98059 and UO126, augment progesterone production in these cell cultures concomitantly with specific elevation of intracellular steroidogenic acute regulatory protein (StAR). Intracellular levels of the cytochrome P450 side-chain cleavage enzyme system do not seem to be affected. Similar observations were made with rat preovulatory or preantral granulosa cells stimulated with LH, FSH, or forskolin. Elevation of StAR expression by the MAPK inhibitors involved elevation of StAR mRNA, as demonstrated by RT-PCR in the human cells. Immunocytochemical studies using specific antibodies to StAR demonstrate a higher content of mitochondrial StAR in control as well as in gonadotropin-stimulated cells in the presence of PD98059 compared with cells not treated with PD98059. The cultured cells express the transcription factor steroidogenic factor-1 (SF-1), the phosphorylation of which is known to activate the expression of StAR, as well as dosage-sensitive sex reversal adrenal hypoplasia congenita, critical region on the X chromosome gene-1 (DAX-1), which is known to negate SF-1 activity. Intracellular levels of DAX-1 decreased significantly during 24 h of incubation of cells with or without LH in the presence of PD98059 or UO126 compared with those in cultures incubated in the absence of the MAPK inhibitors. The expression of SF-1 was suppressed by LH, whereas MAPK inhibitor could block this effect and further elevate SF-1 levels. Thus, activation of the MAPK cascade by gonadotropins may serve as a novel mechanism to down-regulate steroidogenesis via attenuation of StAR expression. Moreover, modulation of DAX-1 and SF-1 intracellular levels in these cells suggests that these transcription factors could be involved in MAPK suppression of StAR expression. PMID- 12727989 TI - Selective beta-cell loss and alpha-cell expansion in patients with type 2 diabetes mellitus in Korea. AB - In the presence of obesity, beta-cell mass needs to be increased to compensate for the accompanying demands and maintain euglycemia. However, in Korea, the majority of type 2 diabetic patients are nonobese. We determined the absolute masses, relative volumes, and ratio of alpha- and beta-cell in the pancreas and islets in normal and diabetic Korean subjects to correlate these findings with the clinical characteristics. Whole pancreases procured from organ donors were divided into 24 parts (control 1, n = 9). Tissue was also obtained by surgical resection after 35 partial pancreatectomies: in 25 diabetic patients, 10 age- and body mass index (BMI)-matched patients of benign or malignant pancreatic tumor without diabetes mellitus (DM) (control 2). Morphometric quantifications were performed. In control 1, the relative volume of beta-cells was 2.1 +/- 0.9%, and the total beta-cell mass was 1.3 +/- 0.3 g. The relative volume of beta-cells was found to be variable (control 1, 2.1 +/- 0.9%; control 2, 1.9 +/- 0.7%; DM, 1.4 +/- 1.0%; P < 0.05 DM vs. control 1 and 2) and showed good correlation with BMI (control 1, r(2) = 0.64; DM, r(2) = 0.55; all subjects, r(2) = 0.38; P < 0.05). Notably, in type 2 diabetic patients, the ratio of alpha-cell area to beta-cell area in the islet was higher than in control 1 and 2 (0.81 +/- 0.4 vs. 0.29 +/- 0.2, 0.20 +/- 0.1, P < 0.05). Additionally, significant alpha-cell expansion and a decreased beta-cell fraction were predominantly observed in larger islets (islet area, >6415 micro m(2); P < 0.05) in control 1 and diabetic patients. The relative volume of beta-cell was found to be correlated with BMI in diabetic patients and normal organ donors. Moreover, decreased beta-cell but increased alpha-cell proportion in the islets suggests for a selective beta-cell loss in the pathogenesis of Korean type 2 diabetes. PMID- 12727990 TI - Sex steroid hormone receptors in human thymoma. AB - In this study we examined the immunohistochemical localization of sex steroid receptors for estrogen alpha (ER alpha) and ER beta, progesterone-A (PR-A) and PR B, and androgen (AR) in human thymoma (n = 132) and correlated these findings with various clinicopathological parameters. We used RT-PCR and real-time PCR to further study the expression of these receptors in 20 thymoma cases. Immunoreactivity for all sex steroid receptors was detected in the nuclei of thymoma epithelial cells. The percentage of immunopositive cases and the H-score values for each receptor (mean +/- SD) were: ER alpha, 66% and 85.8 +/- 80.2; ER beta, 7% and 7.2 +/- 8.7; PR-A, 4% and 2.7 +/- 4.9; PR-B, 49% and 55.8 +/- 68.3; and AR, 15% and 14.1 +/- 11.7, respectively. The results of real-time PCR were consistent with those of immunohistochemistry, especially results for ER alpha, PR-B, and AR. A significant positive correlation was detected between immunoreactivity for ER alpha and PR-B. ER alpha immunoreactivity was inversely correlated with tumor size, clinical stage, WHO classification, and Ki-67 labeling index. In addition, the status of ER alpha immunoreactivity was significantly associated with a better clinical outcome in thymoma patients. Results from our study suggest that estrogens may inhibit thymoma growth via ER alpha, and that ER alpha immunoreactivity may act as a prognostic factor in human thymoma. PMID- 12727991 TI - RAS point mutations and PAX8-PPAR gamma rearrangement in thyroid tumors: evidence for distinct molecular pathways in thyroid follicular carcinoma. AB - A series of 88 conventional follicular and Hurthle cell thyroid tumors were analyzed for RAS mutations and PAX8-PPAR gamma rearrangements using molecular methods and for galectin-3 and HBME-1 expression by immunohistochemistry. A novel LightCycler technology-based method was developed to detect point mutations in codons 12/13 and 61 of the H-RAS, K-RAS, and N-RAS genes. Forty-nine percent of conventional follicular carcinomas had RAS mutations, 36% had PAX8-PPAR gamma rearrangement, and only one (3%) had both. In follicular adenomas, 48% had RAS mutations, 4% had PAX8-PPAR gamma rearrangement, and 48% had neither. Follicular carcinomas with PAX8-PPAR gamma typically showed immunoreactivity for galectin-3 but not for HBME-1, tended to present at a younger patient age and be smaller size, and were almost always overtly invasive. In contrast, follicular carcinomas with RAS mutations most often displayed an HBME-1-positive/galectin-3-negative immunophenotype and were either minimally or overtly invasive. Hurthle cell tumors infrequently had PAX8-PPAR gamma rearrangement or RAS mutations. These results suggest that conventional follicular thyroid carcinomas develop through at least two distinct and virtually nonoverlapping molecular pathways initiated by either RAS point mutation or PAX8-PPAR gamma rearrangement. PMID- 12727992 TI - Progestogenic effects of tibolone on human endometrial cancer cells. AB - Tibolone, a synthetic steroid acting in a tissue-specific manner and used in hormone replacement therapy, is converted into three active metabolites: a Delta(4) isomer (exerting progestogenic and androgenic effects) and two hydroxy metabolites, 3 alpha-hydroxytibolone (3 alpha-OH-tibolone) and 3beta-OH-tibolone (exerting estrogenic effects). In the present study an endometrial carcinoma cell line (Ishikawa PRAB-36) was used to investigate the progestogenic properties of tibolone and its metabolites. This cell line contains progesterone receptors A and B, but lacks estrogen and androgen receptors. When tibolone was added to the cells, complete conversion into the progestogenic/androgenic Delta(4) isomer was observed within 6 d. Furthermore, when cells were cultured with tibolone or when the Delta(4) isomer or the established progestagen medroxyprogesterone acetate was added to the medium, marked inhibition of growth was observed. Interestingly, 3 beta-OH-tibolone also induces some inhibition of growth. These growth inhibitions were not observed in progesterone receptor-negative parental Ishikawa cells, and progestagen-induced growth inhibition of PRAB-36 cells could readily be reversed using the antiprogestagen Org-31489. Upon measuring the expression of two progesterone-regulated genes (fibronectin and IGF-binding protein-3), tibolone, the Delta(4) isomer and medroxyprogesterone acetate showed similar gene expression regulation. These results indicate that tibolone, the Delta(4) metabolite, and to some extent 3 beta-OH-tibolone exert progestogenic effects. Tibolone and most likely 3 beta-OH-tibolone are converted into the Delta(4) metabolite. PMID- 12727993 TI - Ghrelin is involved in the decidualization of human endometrial stromal cells. AB - Successful implantation involves a complex interaction between the endometrium and the embryo. It is well known that several neuropeptides are expressed in the endometrium and placenta during embryonal implantation, suggesting an important role as chemical mediators of the feto-maternal relationship. Ghrelin has recently been identified as the endogenous ligand for the GH secretagogue receptor. Ghrelin is a peptide hormone with many physiological functions, and its expression in the human placenta has been reported. To investigate the involvement of ghrelin in embryonal implantation, we assessed the spatio-temporal expression pattern of ghrelin and its receptor in the human endometrium and placenta through the normal menstrual cycle and in early pregnancy. We also examined the effect of ghrelin on the decidualization of endometrial stromal cells (ESC). Weak expression of ghrelin mRNA was detected in the nonpregnant endometrium, and it was dramatically increased in the decidualized endometrium. A GH secretagogue receptor mRNA was detected in the endometrium throughout the normal menstrual cycle and in early pregnancy, but not in the first trimester placenta. Immunohistochemical analysis using an antighrelin antibody revealed strong signals in decidual cells and extravillous trophoblast cells. Coculture with first trimester placenta up-regulated ghrelin mRNA expression by primary cultured ESC, although sex steroids and 8-bromo-cAMP had no effect. In addition, ghrelin enhanced the decidualization of ESC induced by 8-bromo-cAMP (8-Br-cAMP) in vitro. Thus, ghrelin is a novel paracrine/autocrine factor that is involved in cross-talk between the endometrium and embryo during embryonal implantation. PMID- 12727994 TI - Pituitary tumor transforming gene and fibroblast growth factor-2 expression: potential prognostic indicators in differentiated thyroid cancer. AB - Differentiated thyroid cancers are the most common endocrine cancers, but there are no reliable molecular markers of prognosis. Pituitary tumor transforming gene (PTTG) plays several potential roles in tumor initiation and progression, including regulating mitosis and stimulating expression of fibroblast growth factor (FGF)-2. Increased expression of PTTG has been demonstrated in follicular thyroid lesions, and expression of this oncogene has been identified as a potential prognostic marker in pituitary adenomas and colon carcinomas. We assessed the expression of PTTG and FGF-2 and its receptor FGF-R-1 in 27 differentiated thyroid cancers, and we compared this with expression in 11 normal thyroids, 25 multinodular goiters, and 13 Graves' disease specimens. We also examined the relationship between gene expression and clinical markers of tumor behavior. PTTG and FGF-2 were overexpressed in thyroid carcinomas (9.5-fold increase, P = 0.003, and 5.0-fold increase, P < 0.001, respectively) compared with normal thyroid. Increased FGF-2 mRNA expression was independently associated with the findings of lymph node invasion (R(2) = 0.71; P < 0.001) and distant metastasis (R(2) = 0.55; P = 0.009) at tumor presentation, after taking into account known prognostic factors such as age and gender of the patient and size and type of the tumor. High PTTG expression was independently associated with tumor recurrence (R(2) = 0.64; P = 0.003). We conclude that PTTG and FGF-2 expression are potential prognostic markers (and perhaps therapeutic targets) for differentiated thyroid cancer. PMID- 12727995 TI - Elevated serum soluble vascular endothelial growth factor receptor 1 (sVEGFR-1) levels in women with preeclampsia. AB - Hoping to get more insight into a role of vascular endothelial growth factor (VEGF), a putative substance involved in the development of preeclampsia, we measured concentrations of soluble VEGF receptor-1 (sVEGFR-1), a natural antagonist of VEGF, in serum from women with (n = 31) and without (n = 52) preeclampsia. The concentrations of sVEGFR-1 in serum from women with preeclampsia (median 7791 pg/mL) were > 6-fold higher than those from control (1132 pg/mL, p < 0.0001). The levels of sVEGFR-1 decreased markedly after delivery in both groups. Serum sVEGFR-1 levels of non-preeclamptic women were positively correlated with gestational age (r = 0.570, p < 0.0001), whereas those of preeclamptic women exhibited no correlation with gestational age (r = -0.130, p = 0.476). These findings may point to an involvement of sVEGFR-1 in the pathophysiology of preeclampsia possibly by antagonizing of VEGF effects on the formation of placental vasculature and maternal endothelial cell function. PMID- 12727997 TI - Androgens, estrogens, and bone turnover in men. PMID- 12727998 TI - The role of visual landmarks in the avian familiar area map. AB - The question of whether homing pigeons use visual landmarks for orientation from distant, familiar sites is an unresolved issue in the field of avian navigation. Where evidence has been found, the question still remains as to whether the landmarks are used independent of the map and compass mechanism for orientation that is so important to birds. Recent research has challenged the extent to which experiments that do not directly manipulate the visual sense can be used as evidence for compass-independent orientation. However, it is proposed that extending a new technique for research on vision in homing to include manipulation of the compasses used by birds might be able to resolve this issue. The effect of the structure of the visual sense of the homing pigeon on its use of visual landmarks is also considered. PMID- 12727999 TI - Cadmium disrupts behavioural and physiological responses to alarm substance in juvenile rainbow trout (Oncorhynchus mykiss). AB - Alarm substance is a chemical signal released from fish skin epithelial cells after a predator causes skin damage. When other prey fish detect alarm substance by olfaction, they perform stereotypical predator-avoidance behaviours to decrease predation risk. The objective of this study was to explore the effect of sublethal cadmium (Cd) exposure on the behavioural and physiological responses of juvenile rainbow trout (Oncorhynchus mykiss) to alarm substance. Waterborne exposure to 2 microg Cd l(-1) for 7 days eliminated normal antipredator behaviours exhibited in response to alarm substance, whereas exposures of shorter duration or lower concentration had no effect on normal behaviour. Furthermore, dietary exposure to 3 microg Cd g(-1) in the food for 7 days, which produced the same whole-body Cd accumulation as waterborne exposure to 2 microg l(-1), did not alter normal behaviour, indicating that an effect specific to waterborne exposure alone (i.e. Cd accumulation in the olfactory system) results in behavioural alteration. Whole-body phosphor screen autoradiography of fish exposed to (109)Cd demonstrated that Cd deposition in the olfactory system (rosette, nerve and bulb) during waterborne exposure was greater than in all other organs of accumulation except the gill. However, Cd could not be detected in the brain. A short-term elevation in plasma cortisol occurred in response to alarm substance under control conditions. Cd exposures of 2 microg l(-1) waterborne and 3 microg g(-1) dietary for 7 days both inhibited this plasma cortisol elevation but did not alter baseline cortisol levels. Our results suggest that exposure to waterborne Cd at environmentally realistic levels (2 microg l(-1)) can disrupt the normal behavioural and physiological responses of fish to alarm substance and can thereby alter predator-avoidance strategies, with potential impacts on aquatic fish communities. PMID- 12728000 TI - Male-like behavioral patterns and physiological alterations induced by androgenic gland implantation in female crayfish. AB - The androgenic gland (AG) has been shown to regulate male sexual differentiation and secondary male characteristics in Crustacea. This study presents for the first time in crustaceans evidence for masculinization effects of the AG on reproductive behavior, in addition to morpho-anatomical and physiological effects. AG implantation into immature female red claw crayfish Cherax quadricarinatus inhibited secondary vitellogenesis and development of the ovaries, as well as morphological traits that facilitate maternal egg brooding; it also caused the appearance of secondary male characteristics. However, primary male characteristics and a masculine reproductive system were not developed. In pair encounters, aggression was substantially lower in interactions between AG implanted and intact females than in interactions within AG-implanted or intact pairs. Moreover, elements of mating behavior, i.e. male courtship displays and false copulations, were exhibited by AG-implanted females in several encounters with intact females. In addition to known morpho-anatomical and physiological effects of the AG in crustaceans, the present study suggests that the AG has novel effects on the neural network that generates social behavior. PMID- 12728002 TI - Darkness induces mobility, and saturation deficit limits questing duration, in the tick Ixodes ricinus. AB - The behaviour of Ixodes ricinus nymphs was recorded in 10-day experiments using computer-assisted video-tracking, in the absence of any host stimuli. These ticks switch spontaneously from questing in a desiccating atmosphere to quiescence in a water-saturated atmosphere after dark. Quantification of both questing and quiescence duration demonstrates that questing duration is inversely related to saturation deficit whereas quiescence duration is not. Distance walked after quiescence increased with desiccating conditions, while the distance walked after questing remained unchanged. Almost all locomotor activities of I. ricinus occurred during darkness under either a 14 h:10 h L:D or a 8 h:4 h L:D cycle. We established that all life stages of I. ricinus are equipped to sense shifts in light intensity with bilaterally placed strings of photoreceptors. This permits I. ricinus to use onset of darkness to trigger mobility when desiccation risk is reduced in nature. PMID- 12728001 TI - Metabolic and ionic responses of trout hepatocytes to anisosmotic exposure. AB - Trout hepatocytes exposed to hypo- or hyperosmotic conditions respond by swelling and shrinking, respectively, followed by regulatory volume changes that almost, although not completely, restore cell volume. These anisosmotic conditions have a significant impact on metabolic functions. In hyposmotic medium, oxygen consumption (.VO2) and glucose production rates were significantly reduced, whereas lactate accumulation was not significantly affected. By contrast, hyperosmotic conditions did not affect .VO2 and lactate production but caused a sustained reduction in glucose production. Volume changes were also accompanied by alterations in intracellular free calcium ([Ca2+](i)). At the cell population level, hyposmotic exposure evoked a moderate and slowly developing increase in [Ca2+](i), whereas hyperosmolarity caused a pronounced and sustained increase, which peaked at the time of maximum cell shrinkage but clearly exceeded a mere concentration effect due to volume reduction. Responses of individual cells were highly variable in hyposmotic medium, with only 60% showing a clear increase in [Ca2+](i), while in hyperosmotic conditions all cells displayed elevated [Ca2+](i) levels. A decrease in intracellular pH (pHi) observed in hyposmotic medium was insensitive to EIPA, an inhibitor of Na(+)/H(+) exchange, and SITS, an inhibitor of Cl(-)/HCO(3)(-) exchange, but was prevented in Cl(-)-free medium. In hyperosmotic medium, pHi increased. This alkalinization did not occur under conditions of blocked Na(+)/H(+) exchange and was significantly diminished upon inhibition of Cl(-)/HCO(3)(-) exchange, suggesting an important role of these ion transporters in regulatory volume increase of trout hepatocytes. PMID- 12728003 TI - Comparative analysis of DNA vectors at mediating RNAi in Anopheles mosquito cells and larvae. AB - Heritable RNA interference (RNAi) mediated by transgenes exhibiting dyad symmetry represents an important tool to study the function of genes expressed at late developmental stages. In this study, we determined whether the transcriptional machinery of Anopheles mosquitoes is capable of directing suppression of gene expression from DNA constructs designed to transcribe double-stranded RNA (dsRNA) as extended hairpin-loop RNAs. A series of DNA vectors containing sense and antisense regions of the green fluorescent protein EGFP target gene was developed. The effect of these vectors on a transiently expressed or stably integrated EGFP gene was assessed in an Anopheles gambiae cell line and in Anopheles stephensi larvae. Our data indicate that dsRNA-mediated silencing of a target gene from plasmid DNA can be achieved at high levels in Anopheles cell lines and larvae. The region that links the sense and antisense sequences of the target gene plays a determining role in the degree of silencing observed. These results provide important information for the development of heritable RNAi in Anopheles. PMID- 12728004 TI - Stimulation of JH biosynthesis by the corpora allata of adult female Aedes aegypti in vitro: effect of farnesoic acid and Aedes allatotropin. AB - Previous studies have demonstrated that the synthesis of juvenile hormone (JH) by the isolated corpora allata (CA) complex in vitro as well as the JH titer in the yellow fever mosquito Aedes aegypti are elevated before feeding and low after a blood meal. In the present study, we used an in vitro radiochemical assay to analyze the effect of farnesoic acid (FA) and Aedes allatotropin (Aedes-AT) on the biosynthesis of JH and methyl farnesoate (MF) by the isolated CA complex of A. aegypti adult female. CA complex from day-0 females (0-1 h after emergence) exhibited a low basal juvenile hormone III (JH III) biosynthetic activity and did not respond to either allatotropic or FA stimulation. However, incubation of CA complexes from newly emerged females with Aedes-AT plus FA resulted in very high production of JH III. This is the first report suggesting that allatotropin makes corpora allata in newly emerged females capable for JH biosynthesis. When we studied CA complexes dissected from females 1 day after emergence, the stimulatory action of Aedes-AT was strong and dose-dependent, with maximum stimulation in the range of 10(-8)-10(-9) mol l(-1), suggesting that Aedes-AT is indeed a true allatotropin (a molecule with allatotropic activity) in A. aegypti. The addition to the culture medium of 40 micro mol l(-1) FA, a JH precursor, resulted in a 9-fold increase in JH III biosynthesis in 2-, 4- and 6-day-old sugar-fed females. The two major labeled products synthesized by the stimulated CA complex were identified as JH III and MF by RP-HPLC and GC-MS. Treatment of CA complexes with FA, but not Aedes-AT, resulted in an increase in MF. Application of both Aedes-AT and FA to the CA complexes of 2-, 4- and 6-day-old females resulted in the same effects as FA alone. These data suggest that in sugar-fed females, FA and Aedes-AT exert different effects on the terminal steps in JH biosynthesis. PMID- 12728005 TI - Mechanisms of frequency and amplitude modulation in ring dove song. AB - Birdsong assumes its complex and specific forms by the modulation of phonation in frequency and time domains. The organization of control mechanisms and intrinsic properties causing such modulation have been studied in songbirds but much less so in non-songbirds, the songs of which are often regarded as relatively simple. We examined mechanisms of frequency and amplitude modulation of phonation in ring doves Streptopelia risoria, which are non-songbirds. Spontaneous coo vocalizations were recorded together with concurrent pressure patterns in two different air sacs and air flow rate in the trachea. The results show that amplitude modulation is the result of the cyclic opening and closure of a valve instead of fluctuations in driving pressure, as is the current explanation. Frequency modulation is more complex than previously recognized and consists of gradual, continuous time-frequency patterns, punctuated by instantaneous frequency jumps. Gradual frequency modulation patterns correspond to pressure variation in the interclavicular air sac but not to pressure variation in the cranial thoracic air sac or air flow rate variation in the trachea. The cause of abrupt jumps in frequency has not been identified but can be explained on the basis of intrinsic properties of the vocal organ. Air sac pressure variation as a mechanism for frequency modulation contrasts with the specialized syringeal musculature of songbirds and may explain why the fundamental frequency in non songbird vocalizations is generally modulated within a limited frequency range. PMID- 12728007 TI - Effects of load type (pollen or nectar) and load mass on hovering metabolic rate and mechanical power output in the honey bee Apis mellifera. AB - In this study we tested the effect of pollen and nectar loading on metabolic rate (in mW) and wingbeat frequency during hovering, and also examined the effect of pollen loading on wing kinematics and mechanical power output. Pollen foragers had hovering metabolic rates approximately 10% higher than nectar foragers, regardless of the amount of load carried. Pollen foragers also had a more horizontal body position and higher inclination of stroke plane than measured previously for honey bees (probably nectar foragers). Thorax temperatures ranked pollen > nectar > water foragers, and higher flight metabolic rate could explain the higher thorax temperature of pollen foragers. Load mass did not affect hovering metabolic rate or wingbeat frequency in a regression-model experiment. However, using an analysis of variance (ANOVA) design, loaded pollen and nectar foragers (mean loads 27% and 40% of body mass, respectively) significantly increased metabolic rate by 6%. Mean pollen loads of 18% of body mass had no effect on wingbeat frequency, stroke amplitude, body angle or inclination of stroke plane, but increased the calculated mechanical power output by 16-18% (depending on the method of estimating drag). A rise in lift coefficient as bees carry loads without increasing wingbeat frequency or stroke amplitude (and only minimal increases in metabolic rate) suggests an increased use of unsteady power generating mechanisms. PMID- 12728006 TI - Osmoregulation in an avian nectarivore, the whitebellied sunbird Nectarinia talatala: response to extremes of diet concentration. AB - Water intake of nectarivores is intrinsically linked to nectar concentration. Osmoregulation in whitebellied sunbirds Nectarinia talatala (body mass 9.3+/-0.1 g, mean +/- S.D., N=7), was examined by feeding them sucrose solutions, equivalent to extreme diet concentrations (0.07-2.5 mol l(-1) sucrose; 2-65% w/w), with and without supplementary drinking water. Total water gain was 33-515% of body mass daily. Cloacal fluid (CF) volume increased with diet dilution from 0.4% to 309% of body mass while increases in evaporative water loss (obtained by difference) were also recorded. Osmolality of CF demonstrated the largest scope yet recorded for a bird and was significantly correlated with water flux: mean values were 6-460 mosm kg(-1) H(2)O (minimum 3, maximum 1900 mosm kg(-1)). When supplementary water was provided, its consumption by birds fed concentrated diets (2.5 mol l(-1) sucrose) led to a dramatic reduction in CF osmolality, from 461+/ 253 to 80+/-119 mosm kg(-1) fluid. Sunbirds maintained energy balance on sucrose diets varying tenfold in concentration, from 0.25 to 2.5 mol l(-1); however, on extremely dilute diets (0.07 and 0.1 mol l(-1) sucrose, lower than natural nectar concentrations) their inability to maintain energy balance was probably due to excess preformed water. Total osmotic excretion and concentrations of Na(+) and K(+) increased with high water fluxes, and are a possible physiological constraint for nectarivorous birds on artificial dilute diets devoid of electrolytes. Even low electrolyte levels in nectars may be adequate to replace these losses, but other physiological limitations to the intake of dilute nectars are increased energetic costs of solute recovery, increased heat loss and interference with digestive processes. Sunbirds therefore deal with sugar solutions spanning the range of nectar concentrations by shutting down water excretion on concentrated diets, or, on dilute diets, by producing extremely dilute CF with some of the lowest solute concentrations recorded. PMID- 12728008 TI - Distribution and serotonin-induced activation of vacuolar-type H+-ATPase in the salivary glands of the blowfly Calliphora vicina. AB - Secretory activity in blowfly salivary glands is activated by the hormone serotonin. We have investigated the distribution and activity of two cation pumps that are possibly involved with transepithelial ion transport, i.e. Na(+)/K(+) ATPase and vacuolar-type H(+)-ATPase (V-ATPase). By immunofluorescence labelling of secretory cells, Na(+)/K(+)-ATPase was localized on the basolateral plasma membrane and V-ATPase on the highly folded apical membrane. Activities of both ATPases were probed in salivary gland homogenates by applying specific inhibitors for these ion pumps, namely ouabain and bafilomycin A(1). In control glands, bafilomycin-A(1)-sensitive V-ATPase activity and ouabain-sensitive Na(+)/K(+) ATPase activity accounted for 36% and 19%, respectively, of the total ATPase activity. V-ATPase activity increased approximately twofold after stimulation with serotonin, whereas Na(+)/K(+)-ATPase activity was not significantly affected. Biochemical assays provided evidence that the serotonin-induced activation of V-ATPase activity was accompanied by a recruitment of peripheral V(1) subunits from the cytosol to the plasma membrane, indicative of the assembly of V(0)V(1) holoenzymes. These data show that a V-ATPase located in the apical plasma membranes of the secretory cells is a component of the apical "potassium pump" that has been identified previously by physiological approaches. The V ATPase energizes the apical membrane and provides the primary driving force for fuelling a putative K(+)/nH(+) antiporter and, thus, for fluid secretion. Serotonin-induced assembly of V(0)V(1) holoenzymes might constitute a regulatory mechanism for the control of pump activity. PMID- 12728009 TI - Ectopic transplantation of the accessory medulla restores circadian locomotor rhythms in arrhythmic cockroaches (Leucophaea maderae). AB - The presence of an endogenous circadian clock in the brain of an animal was first demonstrated in the cockroach Leucophaea maderae. However, the clock's cellular basis remained elusive until pigment-dispersing hormone-immunoreactive neurons, which express the clock genes period and timeless in Drosophila, were proposed as pacemaker candidates. In several insect species, pigment-dispersing hormone immunoreactive neurons are closely associated with the accessory medulla, a small neuropil in the optic lobe, which was suggested to be a circadian clock neuropil. Here, we demonstrate that ectopic transplantation of adult accessory medulla into optic lobe-less cockroaches restores circadian locomotor activity rhythms in L. maderae. All histologically examined cockroaches that regained circadian activity regenerated pigment-dispersing hormone-immunoreactive fibres from the grafts to original targets in the protocerebrum. The data show that the accessory medulla is the circadian pacemaker controlling locomotor activity rhythms in the cockroach. Whether pigment-dispersing hormone-immunoreactive neurons are the only circadian pacemaker cells controlling locomotor activity rhythms remains to be examined. PMID- 12728010 TI - Phenotypic flexibility of structure and function of the digestive system of Japanese quail. AB - Organisms adjust their phenotype to fluctuating conditions of the environment and to changing internal demands. We report flexible responses of the gizzard and the small intestine of Japanese quail to a high-fibre diet. Switching from a standard diet to a high-fibre diet results in a highly significant increase in gizzard size, intestine length, mucosal surface, thickness of the intestinal muscular layer and vascularization of the mucosa. After diet switching, increased or decreased gizzard size results from changes in cell size, i.e. smooth muscle cell hypertrophy and hypotrophy, respectively. Increased cell proliferation is not the cause of increase in gizzard size. In the small intestine, however, we found elevated levels of cell proliferation after diet switching and conclude that increased capacity (upregulation) of the small intestine is based on increased rates of mitosis in the intestinal crypts. It is highly probable that elevated levels of cell proliferation in the crypts are balanced by elevated levels of cell extrusion at the tip of intestinal villi. The lipid contents of the liver were reduced, indicating that lipid stores in the liver were mobilized to fuel the flexible response of the gastrointestinal tract. During changes of organ size in response to changes in food composition, resting metabolic rate was not altered. PMID- 12728011 TI - The pathway of myofibrillogenesis determines the interrelationship between myosin and paramyosin synthesis in Caenorhabditis elegans. AB - Examination of null mutants in myosin B and paramyosin yields insights into the complex mechanisms that regulate expression of the three major components of Caenorhabditis elegans body-wall muscle thick filaments myosin A, myosin B and paramyosin. In the absence of myosin B, paramyosin accumulation is reduced, although neither its synthesis nor that of myosin A is affected. This implies that the interaction of myosin B with paramyosin inhibits paramyosin degradation. By contrast, the absence of paramyosin results in reduced synthesis and accumulation of myosin B but has no effect on myosin A synthesis. The non reciprocal effects of the null mutants on turnover and synthesis are best understood as an epigenetic phenomenon that reflects the pathway of thick filament assembly. The synthesis of myosin A and paramyosin, which are involved in the initial steps of thick filament formation, is independent of myosin B; however, a properly assembled paramyosin-containing thick filament core is essential for efficient synthesis of myosin B. PMID- 12728012 TI - Expression of two isoforms of the vacuolar-type ATPase subunit B in the zebrafish Danio rerio. AB - In the present study we tested the hypothesis that two isoforms of the regulatory subunit B of vacuolar-type ATPase (V-ATPase) are expressed in the zebrafish Danio rerio. The complete coding sequences for both isoforms, vatB1 and vatB2, were cloned and sequenced. BLASTX analysis revealed the greatest similarity to amino acid sequences of B subunits from the European eel Anguilla anguilla and rainbow trout Oncorhynchus mykiss. The isoforms were expressed in a bacterial system and the recombinant proteins verified using isoform-specific antibodies directed against vatB isoforms of the eel. The distribution of both isoforms in zebrafish tissues was investigated using reverse transcriptase-polymerase chain reaction and western blot analysis. The results revealed that at the RNA level both isoforms were expressed in all tested organs, i.e. the gills, swimbladder, heart, kidney, liver, spleen, intestine and skeletal muscle. At the protein level, however, there were tissue-specific variations in the levels of the two vatB isoforms expressed. The highest amounts of V-ATPase were detected in total protein preparations from gill, heart and liver tissue. In liver tissue, however, the western blot analysis indicated that vatB1 was not as prominent as vatB2, and immunohistochemistry revealed that antibodies directed against vatB1 yielded a very weak staining in a number of cells, while an antibody directed against vatB1 and vatB2 yielded a strong staining in virtually every cell. Similarly, neurosecretory cells of the small intestine were stained with an antibody directed against vatB1 and vatB2, but not with an antibody specific for vatB1. Therefore we conclude that the differential expression of two isoforms of the V ATPase subunits, which may serve different functions as in several mammalian species, may also be a common phenomenon in teleost fish. PMID- 12728013 TI - Localisation of VIP-binding sites exhibiting properties of VPAC receptors in chromaffin cells of rainbow trout (Oncorhynchus mykiss). AB - The current model for the neuronal control of catecholamine release from piscine chromaffin cells advocates that the neurotransmitters vasoactive intestinal polypeptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP) are co-released with acetylcholine from preganglionic fibres upon nerve stimulation. Both VIP and PACAP elicit the secretion of exclusively adrenaline from rainbow trout chromaffin cells, which presumably arises from the activation of VPAC type receptors. Thus, the goals of the present study were (1) to localise VPAC receptors in the chromaffin cell fraction of the posterior cardinal vein (PCV) of trout and (2) to test the hypothesis that the selective secretion of adrenaline elicited by VIP could be explained by the absence of the VPAC receptors from the noradrenaline-containing cells. Fluorescent labelling of chromaffin cells using aldehyde-induced fluorescence of catecholamines and antisera raised against dopamine beta-hydroxylase (DbetaH) revealed a distinct layer of chromaffin cells lining the walls of the PCV. Furthermore, specific VIP binding sites were demonstrated on chromaffin cells using a biotinylated VIP that was previously established as being bioactive. Although multiple labelling experiments revealed that a number of DbetaH-positive cells were immunonegative for phenylethanolamine N-methyl transferase (PNMT; noradrenaline-containing cells versus adrenaline-containing cells, respectively), labelling of VIP-binding sites was similar to that of DbetaH labelling, suggesting that all chromaffin cells possess VIP-binding sites. Pharmacological assessment of the VIP-binding sites indicated that they exhibited characteristics of VPAC receptors. Specifically, the labelling of VIP-binding sites was prevented after pre-treatment of PCV tissue sections with unlabelled VIP, PACAP or the specific VPAC receptor antagonist VIP 6-28. By contrast, sections pre-treated with the PAC(1) receptor blocker PACAP 6-27 displayed normal labelling of VIP-binding sites. Finally, partial cDNA clones for the trout VPAC(1) and VPAC(2) receptor were obtained and sequenced. Tissue distribution experiments using RT-PCR revealed the presence of VPAC(1) receptor mRNA but not that of the VPAC(2) receptor in the PCV tissue. The results provide direct evidence that VIP and PACAP can elicit the secretion of adrenaline from the chromaffin tissue via specific VIP-binding sites that exhibit properties of VPAC receptors. However, the selective secretion of adrenaline by VIP or PACAP cannot be explained by a lack of VIP-binding sites on the noradrenaline-containing cells. PMID- 12728014 TI - Unconventional ventral attachment of time-depth recorders as a new method for investigating time budget and diving behaviour of seabirds. AB - We tested the use of commercially available electronic time-depth recorders (TDRs) to quantify activities and thus total time budgets of seabirds. This new method involved first fitting TDRs onto the birds' bellies (not on their backs), and, secondly, analysing continuous recordings of temperature, light and pressure to differentiate activities on land and at sea. The birds studied were 12 common guillemots (Uria aalge) rearing chicks at Hornoya, in northern Norway. The method successfully recorded five different activities: at the colony, flying, diving, and resting or active at the sea surface. Overall, common guillemots spent 68% of their time at the colony and 32% at sea. While at sea, the birds spent the majority (77%) of their time at the surface, during which they were active 64% of the time, and rested only 13%. Birds engaged in the costly behaviours of flying and diving for shorter times (11% and 12% of their time at sea, respectively). The method allowed us to differentiate between two types of trips to sea based on the presence (foraging trips: 77% of the total number of trips) or absence (non foraging trips: 23%) of dives. On average, foraging trips lasted 3.2 h, but most trips were shorter (<1 h), during which the mean estimated travel distance from the colony was 11 km. Diving occurred in bouts of 7.7+/-6.6 dives (mean +/- S.D.). The mean maximum dive depth was 10.2+/-7.6 m (deepest dive: 37 m), and the mean dive duration and post-dive intervals were 38.7+/-21.3 s (longest dive: 119 s) and 20+/-12 s, respectively. Direct and indirect evidence suggests that common guillemots had no difficulty in finding food during the study period, and that the TDRs had minimal effects on the birds' behaviour and physiology. The method is easy to use in the field and is applicable to many other flying seabird species; it is therefore an efficient way of collecting information on time budgets and diving behaviour in the context of various ecological and monitoring studies. PMID- 12728015 TI - Predicting metabolic rate from heart rate in juvenile Steller sea lions Eumetopias jubatus. AB - The validity of using heart rate to estimate energy expenditure in free-ranging Steller sea lions Eumetopias jubatus was investigated by establishing whether there is a relationship between heart rate (fH) and oxygen consumption rate ((O(2))) in captive sea lions while swimming and resting. Four trained Steller sea lions (2 males and 2 females; mass 87.4-194.4 kg; age 16 months-3 years) were each equipped with a datalogger and two dorsal subcutaneous electrodes to record electrocardiograms from which fH was calculated. (O(2)) (measured using open circuit respirometry) was simultaneously recorded while the previously fasted animals were at rest within an enclosed dry metabolic chamber or while they swam in an enclosed swim mill against water currents of various speeds (0-1.5 m s( 1)). The mean regression equation describing the relationship between fH (beats min(-1)) and (O(2)) (ml h(-1) kg(-0.60)) for all four animals was (O(2)) =(71.3f(H)+/-4.3)-(1138.5+/-369.6) (means +/- S.E.M.) (r(2)=0.69, P<0.01). The relationship demonstrated between fH and (O(2)) while fasting suggests that heart rate can potentially be used to monitor energy consumption in free-ranging Steller sea lions. However, a short-term feeding experiment revealed no significant increase in heart rate following a 6 kg or 12 kg meal to match the observed increase in rate of oxygen consumption. This suggests that heart rate may not accurately reflect energy consumption during digestion events. Additional research should be conducted to further elucidate how the relationship between heart rate and oxygen consumption is affected by such factors as digestive state, stress and age. PMID- 12728016 TI - Molecular cloning and characterization of the human orthologue of the oppo 1 gene encoding a sperm tail protein. AB - We report here the molecular cloning and characterization of a human orthologue of oppo 1, a mouse gene encoding a male germ-cell-specific sperm tail protein, and the organization of its genomic structure. The mRNA of the human oppo 1 gene (h-oppo 1) was expressed exclusively in the testis, and the 30 kDa protein encoded by the mRNA was detected in human testis and sperm. Immunohistochemical analyses showed that human OPPO 1 protein was localized in the flagellae of ejaculated sperm. A human genomic DNA database search indicated that the h-oppo 1 gene mapped to chromosome 17. The genomic structure of h-oppo 1 showed differences in exon/intron usage, the sequence of the 5'-flanking region, and the first intron was rich in Alu repeats as compared with the mouse oppo 1 gene. Comparison of the two genomic sequences indicated that human oppo 1 has evolved independently, resulting in substantial differences in the genomic structure after the human-mouse split, whereas the sequence of the basic functional unit of the oppo 1 gene seems to have been relatively well conserved. PMID- 12728018 TI - Gene expression profiling of human endometrial receptivity on days LH+2 versus LH+7 by microarray technology. AB - In humans, embryonic implantation and reproduction depends on the interaction of the embryo with the receptive endometrium. To gain a global molecular understanding of human endometrial receptivity, we compared gene expression profiles of pre-receptive (day LH+2) versus receptive (LH+7) endometria obtained from the same fertile woman (n = 5) in the same menstrual cycle in five independent experiments. Biopsies were analysed using the Affymetrix HG-U95A array, a DNA chip containing approximately 12,000 genes. Using the pre-defined criteria of a fold change >/=3 in at least four out of five women, we identified 211 regulated genes. Of these, 153 were up-regulated at LH+7 versus LH+2, whereas 58 were down-regulated. Amongst these 211 regulated genes, we identified genes that were known to play a role in the development of a receptive endometrium, and genes for which a role in endometrial receptivity, or even endometrial expression, has not been previously described. Validation of array data was accomplished by mRNA quantification by real time quantitative fluorescent PCR (Q PCR) of three up-regulated [glutathione peroxidase 3 (GPx-3), claudin 4 (claudin 4) and solute carrier family 1 member 1 (SLC1A1)] genes in independent LH+2 versus LH+7 endometrial samples from fertile women (n = 3) and the three up regulated genes throughout the menstrual cycle (n = 15). Human claudin-4 peaks specifically during the implantation window, whereas GPx-3 and SLC1A1 showed highest expression in the late secretory phase. In-situ hybridization (ISH) experiments showed that GPx-3 and SLC1A1 expression was restricted to glandular and luminal epithelial cells during the mid- and late luteal phase. The present work adds new and important data in this field, and highlights the complexity of studying endometrial receptivity even using global gene-expression analysis. PMID- 12728017 TI - Gene expression regulating epithelial intercellular junction biogenesis during human blastocyst development in vitro. AB - We investigated gene expression associated with trophectoderm epithelial intercellular junction formation in single human embryos at different stages of cleavage using RT-PCR methods based upon magnetic bead separation of polyA+ RNA. Trophectoderm tight junction (TJ) and desmosome biogenesis contribute to intercellular sealing and tissue integrity, critical for vectorial transport and blastocoel cavity formation. Expression of the various genes throughout human preimplantation development showed differing levels of sensitivity of detection; these genes included claudin-1, occludin (TM4+ and TM4 isoforms), ZO-1 (ZO 1alpha+ and ZO-1alpha- isoforms), ZO-2 and JAM (junction adhesion molecule), and the desmosome junction gene, DSC2 (desmocollin 2). Some transcripts appeared to be expressed throughout preimplantation development (claudin-1, JAM, occludin TM4+ and TM4, ZO-1alpha- isoform) while others tended to be expressed preferentially in later cleavage and associated with blastocyst formation (ZO-2, ZO-1alpha+ isoform, DSC-2), illustrating an expression pattern broadly similar to mouse cleavage stages. Human embryo transcript detection was significantly decreased when reverse transcription was performed in solid phase to generate a bead/cDNA transient library rather than after mRNA elution from beads. Transcript detection tended to be positively correlated with embryo morphological grade using the solid phase method. In blastocysts, occludin TM4-, ZO-1alpha+ and DSC2 transcripts were the most susceptible to failure of detection, indicative of low levels of expression which may impact on trophectoderm differentiation competence. Immunoconfocal microscopy analysis of selected adhesion and TJ proteins in human embryos indicated poor membrane assembly compared with mouse blastocysts, which may further affect embryo viability. PMID- 12728019 TI - Akt as a possible intracellular mediator for decidualization in human endometrial stromal cells. AB - To gain an insight into intracellular mechanisms involved in differentiation of human endometrial cells into decidual cells, we examined the presence of Akt, an emerging intracellular mediator in human endometrial stromal cells (ESC). We explored the mechanisms regulating Akt phosphorylation during the process of progesterone-induced decidualization using a primary cell culture system of ESC. Both Akt and phosphorylated Akt (phospho-Akt) were present in ESC. The total Akt level in ESC cultured for 12 days in the absence of ovarian hormones was similar to ESC treated with estradiol (E(2)) at 10 ng/ml, progesterone at 100 ng/ml or E(2) plus progesterone (E(2)progesterone), whereas the levels of phospho-Akt were markedly decreased with progesterone or E(2) progesterone, compared to control cells. Notably, phospho-Akt levels increased during 12 days of culture in parallel with an increase in total Akt in untreated cells. An increase of phospho Akt in the E(2) progesterone-treated cells was marginal. The level of phospho-Akt in E(2) progesterone-treated cells was markedly reduced compared to control cells at all time points examined. Treatment of the cells with 8-bromo-cAMP decreased the amount of phospho-Akt in ESC in as short a period as 15 min, while no discernible change was observed in the untreated cells. Conversely, H89, an inhibitor of protein kinase A (PKA), significantly increased the amount of phospho-Akt. The addition of H-89 reversed the decrease in the level of phospho Akt seen in the cells treated with E(2) progesterone. Thus, we demonstrated the presence of Akt and its phosphorylated form in human ESC. We further suggest that Akt phosphorylation through the cAMP/PKA signal transduction pathway may regulate cellular functions coupled with decidualization. PMID- 12728020 TI - Epithelial expression of matrix metalloproteinase-26 is elevated at mid-cycle in the human endometrium. AB - The human endometrium is a dynamic tissue, which undergoes extensive tissue remodelling during the menstrual cycle. Due to their involvement in such processes, several well-characterized matrix metalloproteinases (MMP) have previously been studied in the endometrium. MMP-26 is a newly described matrilysin. We studied MMP-26 mRNA in 39 normal endometrial samples obtained across the menstrual cycle. Tissue distribution and cycle variation was examined using in-situ hybridization, Northern blot analyis and real time PCR. The probes for Northern blot analysis and real time PCR recognized non-overlapping sequences. MMP-26 was localized exclusively in epithelial cells of both glands and the luminal surface. Expression increased during the proliferative phase to a maximum at mid-cycle, then decreased to non-detectable levels in the late secretory and menstrual phases. Expression of MMP-26 mRNA in endometrial tissue explants in vitro required stimulation with both estradiol and progesterone. The tissue content of c-jun mRNA was assayed, since c-jun, as part of the enhancer complex AP-1, may be involved in regulation of MMP-26 gene transcription. The pattern of c-jun expression over the menstrual cycle was similar to that of MMP 26. Epithelial expression in the peri- and post-ovulatory stages of the menstrual cycle suggests the involvement of MMP-26 in reproductive processes. PMID- 12728022 TI - Human placental GnRH-like factors: II. Inhibition of enzymatic degradation of GnRH-II and [D-Trp6]GnRH ethylamide tracers by human term placental cytosol fractions reveals the presence of GnRH-binding protein(s). AB - We describe the preliminary characterization of GnRH-binding protein(s) in human placental cytosol. Samples were analysed by chromatography on Sephadex G25. Radiolabelled GnRH and its analogues elute significantly later than the total column volume (V(t)) on Sephadex G25 column chromatography. However, incubation of GnRH II or GnRH agonist tracers with human placental cytosol reduced the intact tracer peak, with the concomitant appearance of a new peak eluting in the total column volume (V(t)). This peak increased with increasing cytosol concentration and duration of incubation, and probably represented degraded GnRH tracer, since (i) degradation-resistant GnRH agonist tracer, [D-Trp(6)]GnRH EtA, was inactivated more slowly than GnRH II, (ii) boiling of cytosol fractions abolished formation of this peak and (iii) peptidase inhibitors blocked its formation. A second new tracer peak eluted in the column void volume (V(o)) and was largely unaffected by peptidase inhibitor concentrations that blocked tracer degradation. The magnitude of this high molecular weight peak depended on the GnRH tracer employed, cytosol concentration, and the pH, duration and temperature of incubation. Tracer associated with this third peak appeared similar to intact GnRH tracer by TLC. Unlabelled GnRH analogues and isoforms decreased both tracer degradation and formation of the V(o) peak, but their specificity and affinity for the two processes differed. Ligand blots identified several bands that were abolished by inclusion of unlabelled agonist during incubation. Our data indicate the presence of specific GnRH binding protein(s) and GnRH peptidases that may modulate local actions of GnRH in the human placenta. PMID- 12728021 TI - A novel serine protease of the mammalian HtrA family is up-regulated in mouse uterus coinciding with placentation. AB - This paper characterizes a novel gene, previously identified as uniquely regulated at implantation in mouse uterus. We cloned its full mRNA sequence encoding a serine protease possessing an IGF-binding domain and named it pregnancy-related serine protease (PRSP). PRSP is structurally similar to mammalian HtrA1 (56% amino acid similarity). Northern analysis revealed that the expression of PRSP mRNA was low before pregnancy, but it was increased at implantation and markedly up-regulated post-implantation. In-situ hybridization localized low levels of mRNA expression to the epithelium and stroma during very early pregnancy, but high expression to the decidual cells on day 8.5, primarily at the mesometrial pole where the placenta was forming. By day 10.5, PRSP mRNA was detected in the placenta. We also cloned an alternatively spliced PRSP mRNA that is expressed at a very low level. We located PRSP gene on chromosome 5 and established its intron/exon structure, which unambiguously explains how the two mRNA variants are produced through alternative splicing. Based on PRSP protein domain structure and its unique expression during pregnancy, we propose that PRSP plays an important role in the formation/function of the placenta. PMID- 12728023 TI - An evaluation of PGD in clinical genetic services through 3 years application for prevention of beta-thalassaemia major and sickle cell thalassaemia. AB - PGD represents an alternative within prenatal diagnosis services, which avoids terminating affected on-going pregnancies. In Greece, prevention programmes for haemoglobinopathies, including the option of prenatal diagnosis, are well established. Following optimization of a single-cell genotyping strategy (designed to be applicable for the majority of beta-thalassaemia major or sickle thalassaemia genotype interactions) along with close collaboration with an IVF unit, we integrated the option of PGD for at-risk couples with a problematic reproductive history. A total of 59 couples requesting PGD were counselled, of whom 41 initiated 63 PGD cycles. Following standard assisted reproduction treatment for oocyte retrieval, 20 cycles were cancelled (too few oocytes and/or poor quality embryos), but in 43 cycles single blastomeres were biopsied from 3 day embryos and genotyped (total 302). Diagnosis was achieved for 236 embryos, and 100 of 125 unaffected embryos were transferred. Sixteen pregnancies were established, although six were lost within the first trimester. Ten pregnancies underwent second trimester prenatal diagnosis, with nine pregnancies (13 babies: six singletons, two twins and one triplet) confirmed unaffected, although one singleton was a PGD misdiagnosis and terminated. The triplet pregnancy was selectively reduced to twins, and nine pregnancies went to term, with 12 healthy babies born. This report highlights advantages, limitations and approaches towards improvement when incorporating PGD within genetic services for a common recessive disease. PMID- 12728024 TI - Operative compared with nonoperative treatment of a thoracolumbar burst fracture without neurological deficit. A prospective, randomized study. AB - BACKGROUND: To our knowledge, a prospective, randomized study comparing operative and nonoperative treatment of a thoracolumbar burst fracture in patients without a neurological deficit has never been performed. Our hypothesis was that operative treatment would lead to superior long-term clinical outcomes. METHODS: From 1994 to 1998, forty-seven consecutive patients (thirty-two men and fifteen women) with a stable thoracolumbar burst fracture and no neurological deficit were randomized to one of two treatment groups: operative (posterior or anterior arthrodesis and instrumentation) or nonoperative treatment (application of a body cast or orthosis). Radiographs and computed tomography scans were analyzed for sagittal alignment and canal compromise. All patients completed a questionnaire to assess any disability they may have had before the injury, and they indicated the degree of pain at the time of presentation with use of a visual analog scale. The average duration of follow-up was forty-four months (minimum, twenty-four months). After treatment, patients indicated the degree of pain with use of the visual analog scale and they completed the Roland and Morris disability questionnaire, the Oswestry back-pain questionnaire, and the Short Form-36 (SF 36) health survey. RESULTS: In the operative group (twenty-four patients), the average fracture kyphosis was 10.1 degrees at the time of admission and 13 degrees at the final follow-up evaluation. The average canal compromise was 39% on admission, and it improved to 22% at the final follow-up examination. In the nonoperative group (twenty-three patients), the average kyphosis was 11.3 degrees at the time of admission and 13.8 degrees at the final follow-up examination after treatment. The average canal compromise was 34% at the time of admission and improved to 19% at the final follow-up examination. On the basis of the numbers available, no significant difference was found between the two groups with respect to return to work. The average pain scores at the time of the latest follow-up were similar for both groups. The preinjury scores were similar for both groups; however, at the time of the final follow-up, those who were treated nonoperatively reported less disability. Final scores on the SF-36 and Oswestry questionnaires were similar for the two groups, although certain trends favored those treated without surgery. Complications were more frequent in the operative group. CONCLUSION: We found that operative treatment of patients with a stable thoracolumbar burst fracture and normal findings on the neurological examination provided no major long-term advantage compared with nonoperative treatment. PMID- 12728025 TI - Gender differences in muscular protection of the knee in torsion in size-matched athletes. AB - BACKGROUND: Female athletes who participate in sports involving jumping and cutting maneuvers are up to eight times more likely to sustain a rupture of the anterior cruciate ligament than are men participating in the same sports. We tested the hypothesis that healthy young women are able to volitionally increase the apparent torsional stiffness of the knee, by maximally activating the knee muscles, significantly less than are size-matched men participating in the same type of sport. METHODS: Twenty-four NCAA (National Collegiate Athletic Association) Division-I athletes (twelve men and twelve women) competing in sports associated with a high risk of injury to the anterior cruciate ligament (basketball, volleyball, and soccer) were compared with twenty-eight collegiate endurance athletes (fourteen men and fourteen women) participating in sports associated with a low risk of such injuries (bicycling, crew, and running). Male and female pairs were matched for age, height, weight, body mass index, shoe size, and activity level. Testing was performed with a weighted pendulum that applied a medially directed 80-N impulse force to the lateral aspect of the right forefoot. The resulting internal rotation of the leg was measured optically, to the nearest 0.25 degrees, at 30 degrees and 60 degrees of knee flexion, both with and without maximal activation of the knee muscles. RESULTS: Maximal rotations of the leg were greater in women than in men in both the passive and the active muscle state (16% and 27% greater [p = 0.01 and p = 0.02], respectively). Moreover, female athletes exhibited a significantly (18%) smaller volitional increase in apparent torsional stiffness of the knee under internal rotation loading than did the matched male athletes (p = 0.014); this was particularly the case for those who participated in sports involving jumping and pivoting maneuvers (42% difference between genders, p = 0.001). CONCLUSIONS: The collegiate female athletes involved in high-risk sports exhibited less muscular protection of the knee ligaments during external loading of the knee than did size and sport-matched male athletes. PMID- 12728026 TI - Midshaft malunions of the clavicle. AB - BACKGROUND: The purpose of this study was to analyze the functional results of corrective osteotomy of a malunited clavicular fracture in patients with chronic pain, weakness, neurologic symptoms, and dissatisfaction with the appearance of the shoulder. METHODS: We identified fifteen patients (nine men and six women with a mean age of thirty-seven years) who had a malunion following nonoperative treatment of a displaced midshaft fracture of the clavicle. The mean time from the injury to presentation was three years (range, one to fifteen years). Outcome scores revealed major residual deficits. The mean amount of clavicular shortening was 2.9 cm (range, 1.6 to 4.0 cm). All patients underwent corrective osteotomy of the malunion through the original fracture line and internal fixation. RESULTS: At the time of follow-up, at a mean of twenty months (range, twelve to forty-two months) postoperatively, the osteotomy site had united in fourteen of the fifteen patients. All fourteen patients expressed satisfaction with the result. The mean DASH (Disabilities of the Arm, Shoulder and Hand) score for all fifteen patients improved from 32 points preoperatively to 12 points at the time of follow-up (p = 0.001). The mean shortening of the clavicle improved from 2.9 to 0.4 cm (p = 0.01). There was one nonunion, and two patients had elective removal of the plate. CONCLUSIONS: Malunion following clavicular fracture may be associated with orthopaedic, neurologic, and cosmetic complications. In selected cases, corrective osteotomy results in a high degree of patient satisfaction and improves patient-based upper-extremity scores. PMID- 12728027 TI - Factors influencing the development of osteonecrosis in patients treated for slipped capital femoral epiphysis. AB - BACKGROUND: Osteonecrosis is a serious complication of the treatment of slipped capital femoral epiphysis. The purpose of the present study was to identify factors influencing the development of osteonecrosis. METHODS: Two hundred and forty patients who had been treated for slipped capital femoral epiphysis between 1965 and 1999 were retrospectively evaluated. Treatment included stabilization with a spica cast or fixation with one to four pins or screws. Radiographs that had been made at the time of presentation, before and after the operation, and at consecutive follow-up examinations were reviewed. Osteonecrosis was defined retrospectively on the basis of radiographic evidence of sclerosis and collapse of the femoral head. The risk of development of osteonecrosis was correlated with various clinical and radiographic parameters. RESULTS: All twenty-one patients in whom osteonecrosis developed had presented with an unstable slipped capital femoral epiphysis. None of the 204 patients who had presented with a stable slipped capital femoral epiphysis, regardless of grade, had development of osteonecrosis. In the group of patients who had presented with an unstable slipped capital femoral epiphysis, the risk of development of osteonecrosis increased with the severity (grade) of the slip. Osteonecrosis was more likely to develop in patients who had been treated with multiple pins than in those who had been treated with a single cannulated screw. CONCLUSIONS: Patients who have a stable slipped capital femoral epiphysis are not at risk for the development of osteonecrosis when treated with pinning in situ. Patients who have an unstable slipped capital femoral epiphysis have a decreased risk of osteonecrosis when treated with pinning in situ. Complete or partial reduction of an unstable slipped capital femoral epiphysis increases the risk of development of osteonecrosis. Pinning in situ without reduction with a single cannulated screw is the method of choice for the treatment of a slipped capital femoral epiphysis. PMID- 12728028 TI - Effect of humeral condylar resection on strength and functional outcome after semiconstrained total elbow arthroplasty. AB - BACKGROUND: Under certain conditions it is standard practice to excise ununited humeral condyles during insertion of a semiconstrained total elbow prosthesis. Since the osseous origins of the common extensors and flexor-pronator muscles are lost, it has been postulated that this excision has a negative effect on strength. We are not aware of any previous study in which this issue has been investigated with use of standardized, objective testing of muscle strength. METHODS: We used objective testing to determine the effect of condylar resection on the muscle strength of the elbow, forearm, wrist, and hand in thirty-two patients who had undergone total elbow arthroplasty. To eliminate bias, the normal, contralateral limb served as the control, and all strength values are given as a percentage of the normal side. The humeral condyles were intact in sixteen patients and had been resected in the other sixteen. Patient demographics were similar in the two groups. RESULTS: There were no significant differences between the two groups with regard to strength of pronation (103% of the normal side in the group with intact condyles compared with 89% in the group with resection of the condyles; p = 0.40), supination (68% compared with 89%; p = 0.49), wrist flexion (66% compared with 56%; p = 0.46), wrist extension (75% compared with 65%; p = 0.40), or grip strength (83% compared with 72%; p = 0.40). There was also no difference between the two groups with regard to the Mayo Elbow Performance Score (79 points in the group with intact condyles compared with 77 points in the group with resection of the condyles; p = 0.67). CONCLUSIONS: Condylar resection has a minimal, clinically irrelevant effect on forearm, wrist, and hand strength and no effect on the Mayo Elbow Performance Score following total elbow arthroplasty. Thus, the findings of our study support the practice of condylar resection, which simplifies total elbow arthroplasty for many conditions. PMID- 12728029 TI - Treatment of the dysplastic acetabulum with Wagner spherical osteotomy. A study of patients followed for a minimum of twenty years. AB - BACKGROUND: The purpose of this study was to evaluate the long-term clinical and radiographic results of spherical acetabular osteotomy, performed with the surgical technique described by Wagner, in patients with hip dysplasia. METHODS: The results of the first twenty-two spherical osteotomies performed by one surgeon at one institution were reviewed at a minimum of twenty years (median, 23.9 years; maximum, 29.3 years) postoperatively. Preoperative and follow-up radiographic measurements included the lateral and anterior center-edge angles, acetabular index angle, and acetabulum-head index of Heyman and Herndon. Anteroposterior radiographs of the pelvis were evaluated for the presence of joint congruency, joint-space narrowing, increased sclerosis of the subchondral bone, and bone cysts. Clinical evaluation was performed with use of the Harris hip score. RESULTS: Osteotomy improved the mean lateral center-edge angle from -2 degrees to +13 degrees and the mean acetabulum-head index from 52% to 72%. The mean postoperative anterior center-edge angle was 23 degrees (range, -1 degrees to 62 degrees ). Seven (32%) of the twenty-two hips were converted to a total hip replacement. At the latest follow-up examination, the average Harris hip score of the remaining fifteen patients was 86 points (range, 50 to 100 points). The clinical result was rated good or excellent for eleven of the fifteen patients. At the latest follow-up examination, the severity grade of the osteoarthritis was unchanged in thirteen hips. Only three of the nine hips that subsequently required a total hip replacement or that showed progressive osteoarthritis had been congruent after the index operation, whereas ten of the thirteen hips that did not require total hip replacement or show progressive osteoarthritis had been congruent after the index operation. The twenty-year Kaplan-Meier survival estimate, with conversion to total hip replacement as the end point, was 86.4% (95% confidence interval, 63.4% to 95.4%). The twenty-five-year survival estimate was 65.1% (95% confidence interval, 35.6% to 83.7%). CONCLUSIONS: The Wagner spherical osteotomy prevented progression of osteoarthritis both clinically and radiographically in a high proportion of patients with residual hip dysplasia who were followed for a minimum of twenty years. Operative restoration of joint congruency is associated with a satisfactory long-term outcome in a very high proportion of cases. PMID- 12728030 TI - Patient compliance in avoiding wrong-site surgery. AB - BACKGROUND: Wrong-site orthopaedic surgery is an uncommon, devastating, and preventable complication. The sole responsibility for avoiding this inadvertent event has historically been placed on physicians, nurses, and ancillary health care personnel. Very little attention has been focused on the role of the patient. The successful outcome of any surgical or medical intervention requires an interactive doctor-patient relationship. The hypothesis of this study was that a substantial number of patients who undergo elective orthopaedic surgery do not comply with instructions designed specifically to prevent wrong-site surgery. METHODS: We prospectively evaluated the frequency with which 100 consecutive patients in a private foot-and-ankle practice followed the explicit preoperative instruction, before they underwent elective orthopaedic surgery, to mark "NO" on the extremity that was not to be operated on. Full compliance was defined as a mark on the correct extremity consistent with the instructions. Partial compliance was defined as a mark that was different from that requested by the specific preoperative instructions, and noncompliance was defined as the absence of any mark. Specific demographic and surgical factors were recorded from medical charts and compared between compliant and noncompliant patients. RESULTS: Fifty nine of the 100 patients marked the extremity correctly, thirty-seven made no mark, and four were considered partially compliant. Of the ten patients with a Workers' Compensation claim, seven were noncompliant compared with thirty (33%) of the ninety patients who had not made a Workers' Compensation claim (p = 0.023). Patients who had had a previous related surgical procedure also had a significantly higher rate of noncompliance (51%; nineteen of thirty-seven) compared with those with no previous surgery (29%; eighteen of sixty-three; p = 0.023). CONCLUSIONS: A surprisingly high number of patients do not comply with explicit preoperative instructions created specifically to prevent wrong-site surgery. This behavior suggests that patients expect the system to "take care of everything," despite solicitation of their active participation to avoid such adverse events. Although physicians and related health-care personnel certainly have the greatest responsibility to provide the highest possible quality of care, patients undergoing surgery must be encouraged to take a more active role in their health care in order to optimize outcome and minimize risk. PMID- 12728031 TI - Foot and ankle fractures in elderly white women. Incidence and risk factors. AB - BACKGROUND: Although foot and ankle fractures are among the most common nonspinal fractures occurring in older women, little is known about their epidemiology. This study was designed to determine the incidence of and risk factors for foot and ankle fractures in a cohort of 9704 elderly, nonblack women enrolled in the multicenter Study of Osteoporotic Fractures. METHODS: At their first clinic visit, between 1986 and 1988, the women provided information regarding lifestyle, subjective health, and function. Bone mineral density was measured in the proximal and distal parts of the radius and in the calcaneus. The women were followed for a mean of 10.2 years, during which time 301 of them had a foot fracture and 291 had an ankle fracture. The fractures were classified with use of a modification of the Orthopaedic Trauma Association's guidelines. RESULTS: The incidence of foot fractures was 3.1 per 1000 woman-years, and the incidence of ankle fractures was 3.0 per 1000 woman-years. The most common ankle fracture was an isolated fibular fracture (prevalence, 57.6%), and the most common foot fracture was a fracture of the fifth metatarsal (56.9%). Women who sustained an ankle fracture had been slightly younger at the time of study enrollment than the women who did not sustain such a fracture (71.0 compared with 71.7 years), they had a higher body mass index (27.6 compared with 26.5), and they were more likely to have fallen within the twelve months prior to filling out the original questionnaire (38.1% compared with 29.7%). The appendicular bone mineral density was not significantly different between these two groups of subjects. Women who sustained a foot fracture had a lower bone mineral density in the distal part of the radius (0.345 g/cm (2) compared with 0.363 g/cm (2) ) and a lower calcaneal bone mineral density (0.394 g/cm (2) compared with 0.404 g/cm (2) ) than the women without a foot fracture, they were less likely to be physically active (62.3% compared with 67.8%), and they were more likely to have had a previous fracture after the age of fifty (45.5% compared with 36.8%) and to be using either long or short-acting benzodiazepines. CONCLUSIONS: Overall, foot fractures appeared to be typical osteoporotic fractures, whereas ankle fractures occurred in younger women with a relatively high body mass index. PMID- 12728032 TI - Revision total hip arthroplasty performed after fracture of a ceramic femoral head. A multicenter survivorship study. AB - BACKGROUND: The alumina ceramic femoral head was introduced for total hip arthroplasty approximately thirty years ago. One of its main drawbacks was the risk of implant fracture. The aim of this study was to examine the results of revision total hip replacement performed specifically to treat a fracture of a ceramic femoral head and to identify technical factors that affected the outcomes. METHODS: One hundred and five surgical revisions to treat a fracture of a ceramic femoral head, performed at thirty-five institutions, were studied. The patients were examined clinically by the operating surgeon at the time of the last follow-up. The surgeon provided the latest follow-up radiographs, which were compared with the immediate postoperative radiographs. The success of the revisions was assessed with Kaplan-Meier survivorship analysis, with the need for repeat revision as the end point. We evaluated the complication rate and radiographic changes indicative of implant loosening. The average duration of follow-up between the index revision and the last clinical and radiographic review was 3.5 years (range, six months to twenty years). RESULTS: Following the revisions, radiographic evidence of cup loosening was seen in twenty-two hips (21%) and radiographic evidence of femoral loosening was seen in twenty-two (21%). One or several repeat revisions were necessary in thirty-three patients (31%) because of infection (four patients), implant loosening (twenty), osteolysis (eight), or fracture of the revision femoral head component (one). The survival rate at five years was 63% (95% confidence interval, 51% to 75%). The survival rate was significantly worse when the cup had not been changed, when the new femoral head was made of stainless steel, when a total synovectomy had not been done, and when the patient was less than fifty years old. CONCLUSIONS: Fracture of a ceramic femoral head component is a rare but potentially serious event. A suitable surgical approach, including total synovectomy, cup exchange, and insertion of a cobalt-chromium or new ceramic femoral ball minimizes the chance of early loosening of the implants and the need for one or more repeat revisions. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series [no, or historical, control group]). PMID- 12728033 TI - Cyclic loading of olecranon fracture fixation constructs. AB - BACKGROUND: Despite the good results that are usually reported after fixation at the sites of olecranon fractures and osteotomies, problems such as loss of fixation, nonunion, and the need for revision surgery are still encountered. Various types of fixation have been recommended, but few have been evaluated with use of clinically relevant cyclic load testing at appropriate levels of stress. The purpose of the present study was to test multiple olecranon fixation techniques under physiologic cyclic loads. METHODS: We studied ten cadaveric elbows with use of cyclic loading that simulated (1) active range of motion and (2) pushing up from a chair. Each specimen underwent fixation of a simulated 50% transverse olecranon fracture with use of intramedullary and cortically fixed tension band constructs (in randomized order) followed by fixation with a 7.3-mm diameter cancellous screw with and without a tension band. Displacement transducers were placed posteriorly on the tension side and anteriorly near the articular surface. RESULTS: Both configurations involving the 7.3-mm-diameter cancellous screw provided the most stable fixation-nearly five times better than that provided by the Kirschner-wire techniques. Use of the tension band in conjunction with the intramedullary screw improved the stability of fixation. In none of the constructs did the AO tension band result in compression across the osteotomy gap. CONCLUSIONS AND CLINICAL RELEVANCE: The use of a 7.3-mm screw in conjunction with a tension band provided better fixation of simulated displaced transverse fractures than did the use of Kirschner wires in conjunction with a tension band or the use of a screw only. The AO principle of converting posterior tensile forces to articular compressive forces was not demonstrated in this study. We therefore question the validity of the tension band concept in olecranon fracture fixation and recommend passive rather than active range of motion in the immediate postoperative period to limit fracture distraction. PMID- 12728035 TI - Operative release of complete ankylosis of the elbow due to heterotopic bone in patients without severe injury of the central nervous system. AB - BACKGROUND: Although uncommon, complete ankylosis of the elbow secondary to heterotopic ossification results in severe disability. The results of surgical management remain unclear. METHODS: A single surgeon used a consistent operative technique to treat complete osseous ankylosis of the elbow in eleven limbs in seven patients after severe burns and in nine elbows in eight patients after trauma. The elbows in the burn cohort were more often ankylosed in extension (average, 47 degrees of flexion) compared with those in the trauma cohort (66 degrees of flexion), and they had more skin problems (three elbows required a free microvascular muscle transfer for coverage) and associated problems of the shoulder, wrist, and hand. RESULTS: Four patients in the burn cohort and three patients in the trauma cohort failed to regain at least 80 degrees of ulnohumeral motion. After a repeat release in three burn patients and three trauma patients, and at an average follow-up of forty months, the average arc of ulnohumeral motion was 81 degrees in the burn cohort and 94 degrees in the trauma cohort. Six of the eleven limbs in the burn cohort and five of the nine in the trauma cohort had a good result. The average score according to the American Shoulder and Elbow Surgeons elbow assessment form was 72 points for the burn cohort and 76 points for the trauma cohort. CONCLUSIONS: Osseous ankylosis of the elbow is a severely disabling problem, and attempts to regain mobility are both worthwhile and safe. The results are comparable when the ankylosis is caused by burns or trauma despite the greater complexity of osseous ankylosis in the burned arm. Patients and surgeons should be aware of the small risk of recurrent heterotopic ossification and the moderate risk of pain or recurrent contracture after operative release. PMID- 12728034 TI - Gradual femoral lengthening with the Albizzia intramedullary nail. AB - BACKGROUND: Gradual limb lengthening with currently used external fixation techniques can result in less than optimal outcomes, with complications including infection, stiffness of adjacent joints, and secondary axial deviation of the extremity. We describe a totally implantable lengthening device designed to provide results similar to those achieved with external fixation devices, with fewer complications and improved outcomes. METHODS: Between 1993 and 1997, thirty one patients (forty-one femora) underwent limb lengthening with a new internal fixation technique (Albizzia) to treat a congenitally short extremity (thirteen patients), post-traumatic limb-length inequality (eleven patients), or developmental problems (seven patients). Twenty-one patients (twenty-one femora) underwent unilateral surgery to equalize the limb lengths, and ten (twenty femora) underwent bilateral surgery to correct short stature. The mean age was twenty years and one month (range, twelve to thirty-nine years). After intramedullary corticotomy of the diaphysis of the femur, an intramedullary nail was inserted in an antegrade fashion. Fifteen alternating internal and external rotation maneuvers of the lower limb elongated the nail by 1 mm. The outcomes were assessed clinically and radiographically at a mean of fifty months postoperatively. RESULTS: The gain in femoral length averaged 3.4 cm (range, 2 to 5.5 cm) after the unilateral lengthening procedures and 6.3 cm (range, 4.6 to 8.4 cm) after the bilateral procedures. Patients underwent an average of three operations on each limb; these procedures included, in addition to the nail insertion and nail removal, ratcheting under general anesthesia in thirteen limbs and eleven procedures to treat complications in nine patients. At the time of follow-up, no patient had axial deviation of the limb secondary to lengthening. CONCLUSIONS: Femoral lengthening with use of the minimally invasive Albizzia technique provides a reasonable alternative to external fixation that is well tolerated by patients and results in excellent function with little or no distortion of body image. PMID- 12728036 TI - Effects of tensioning errors in split transfers of tibialis anterior and posterior tendons. AB - BACKGROUND: Split transfers of the tibialis anterior and posterior tendons are commonly performed to address hindfoot varus deformities in patients with cerebral palsy, stroke, or brain injury. Poor outcomes from these procedures are often attributed to a failure to tension the transferred tendon properly, but the mechanical effects of this aspect of the procedure have not been quantified, to our knowledge. The purpose of the present study was to use a cadaver model to examine changes in the actions of these muscles that occur when the tensions in the halves of the split tendon are intentionally balanced or unbalanced to varying degrees. METHODS: Tendon excursion was measured in seven cadaveric specimens before and after split tendon transfer with experimentally controlled tensions in the halves of the split tendon. The muscle moment arm, a quantitative indicator of the action of a muscle about a joint axis, was calculated as the derivative of tendon excursion with respect to the subtalar joint angle. RESULTS: The tibialis anterior had an eversion moment arm with the subtalar joint in a neutral position following surgery, but the tibialis posterior had virtually no action in the neutral position. Following the split transfers with ideally balanced tension, subtalar joint rotations of >5 degrees strongly influenced the moment arm of the tibialis posterior (p < 0.0002), indicating that its action depends on the position of the hindfoot. The moment arm of the tibialis anterior, however, was influenced only by rotations of >/=20 degrees (p > 0.1741 for each angle pair comparison of <20 degrees ). Moment arms were generally insensitive to imbalances in tension between the medial and lateral tendon halves; significant differences in the moment arm (p < 0.05), compared with that in the balanced condition, were seen only when one half was slack or nearly so. CONCLUSIONS: These results suggest that it is possible for a split tendon transfer to be successful over a large range of tensionings. Split transfer of the tibialis posterior tendon produced the desired mechanical outcome in that the tibialis posterior had an eversion moment arm when the foot was inverted and an inversion moment arm when the foot was everted. Split transfer of the tibialis anterior to the cuboid, however, produced a muscle that consistently functioned as an everter regardless of the position of the hindfoot. PMID- 12728037 TI - Thoracoscopic intradiscal spine fusion using a minimally invasive gene-therapy technique. AB - BACKGROUND: Gene therapy has been utilized to achieve posterior intertransverse process fusion in rodents. To our knowledge, however, no one has previously reported on the use of this technique to achieve anterior spinal fusion in mammals. The purpose of the present study was to determine if a gene-therapy technique can be utilized to achieve anterior intradiscal fusion in pigs with use of minimally invasive techniques. METHODS: Mesenchymal stem cells were isolated from each of three pigs, expanded in culture, and transduced with an adenovirus carrying either the gene for bone morphogenetic protein-2 (Adv-BMP2) or the control gene, beta-galactosidase (Adv-betagal). In vitro, assays were performed to detect BMP-2 expression as well as protein markers of bone formation. In vivo, four thoracic disc spaces in each of three pigs were injected thoracoscopically with cells after 1 cm (3) of the disc had been removed. In each of the three pigs, two discs were injected with autologous mesenchymal stem cells transduced with Adv-BMP2, the third disc was injected with cells transduced with Adv-betagal (control 1), and the fourth disc served as the sham-operated control (control 2). The three animals were killed six weeks after the implantation. Computerized tomographic scanning was performed on two of the specimens, and histological examination was performed on all specimens. The computerized tomographic scans and histological examinations were then interpreted in a blinded fashion. RESULTS: In the in vitro study, a human BMP-2 protein band was detected in the medium of Adv-BMP2-transduced stem cells but not in that of the control cells. The Adv-BMP2-transduced stem cells were associated with a fivefold increase in alkaline phosphatase activity compared with the controls as well as with matrix mineralization and increased protein expression of type-I collagen, osteopontin, and bone sialoprotein. In the in vivo study, radiographic examination demonstrated anterior spinal fusion in all six disc spaces that had been treated with implantation of Adv-BMP2-transduced stem cells. In contrast, the six control disc spaces had little or no intervening bone. Histological examination demonstrated bridging bone from end plate to end plate in all six disc spaces that had been treated with implantation of Adv-BMP2-transduced stem cells. The six control disc spaces had no bridging bone. CONCLUSION: The Adv-BMP2-transduced mesenchymal stem cells produced BMP-2 protein. Further, the cells differentiated into osteoblasts and induced anterior spinal fusion in six of six disc spaces in this pig model. Although many technical and practical challenges remain, the results of the present study suggest that it may eventually be possible to use similar techniques to achieve anterior spinal fusion in humans. PMID- 12728038 TI - Risk factors for Plantar fasciitis: a matched case-control study. AB - BACKGROUND: Plantar fasciitis is one of the more common soft-tissue disorders of the foot, yet little is known about its etiology. The purpose of the present study was to use an epidemiological design to determine whether risk factors for plantar fasciitis could be identified. Specifically, we examined the risk factors of limited ankle dorsiflexion with the knee extended, obesity, and time spent weight-bearing. METHODS: We used a matched case-control design, with two controls for each patient. The matching criteria were age and gender. We identified fifty consecutive patients with unilateral plantar fasciitis who met the inclusion criteria. The data that were collected included height, weight, whether the subject spent the majority of the workday weight-bearing, and whether the subject was a jogger or runner. We used a reliable goniometric method to measure passive ankle dorsiflexion bilaterally. The main outcome measure was the adjusted odds ratio of plantar fasciitis associated with varying degrees of limitation of ankle dorsiflexion, different levels of body mass, and the subjects' reports on weight bearing. RESULTS: Individuals with 10 degrees of ankle dorsiflexion. Individuals who had a body-mass index of >30 kg/m (2) had an odds ratio of 5.6 (95% confidence interval, 1.9 to 16.6) when compared with the referent group of individuals who had a body-mass index of 30 kg/m (2) are also at increased risk for the development of plantar fasciitis. Reduced ankle dorsiflexion, obesity, and work-related weight-bearing appear to be independent risk factors for plantar fasciitis. Reduced ankle dorsiflexion appears to be the most important risk factor. PMID- 12728040 TI - Ten-year results with hydroxyapatite-coated total hip femoral components in patients less than fifty years old. A concise follow-up of a previous report. AB - We report the results of total hip arthroplasty with use of a proximally hydroxyapatite-coated femoral component after a minimum follow-up of ten years in a group of patients who were less than fifty years old at the time of the primary procedure. In the five years since the original publication of our study, two additional stems have undergone revision. Thus, a total of six stems have been revised. A small amount of erosive scalloping of the proximal part of the femur was seen in nearly one-half of the hips; however, all unrevised stems were radiographically stable and no hip had intramedullary osteolysis. The revision rate because of aseptic loosening of the stem was 0.9%, which compares favorably with that for other stems and other fixation methods in young patients at this point in time. This stem is currently being paired with a highly cross-linked polyethylene liner because of cup failures and the need for reoperation secondary to excessive polyethylene wear and proximal femoral osteolysis. PMID- 12728039 TI - Glenoid rim morphology in recurrent anterior glenohumeral instability. AB - BACKGROUND: Knowledge regarding the morphology of the glenoid rim is important when patients with recurrent anterior glenohumeral instability are assessed. Ordinary imaging techniques are not always sensitive enough to demonstrate the morphology of the glenoid rim accurately. We developed a method of three dimensionally reconstructed computed tomography with elimination of the humeral head to evaluate glenoid morphology. The purpose of the present study was to quantify glenoid osseous defects and to define their characteristics in patients with recurrent anterior instability. METHODS: The morphology of the glenoid rim in 100 consecutive shoulders with recurrent unilateral anterior glenohumeral instability was evaluated on three-dimensionally reconstructed computed tomography images with the humeral head eliminated. The configuration of the glenoid rim was evaluated on both en face and oblique views. Concurrently, we also investigated seventy-five normal glenoids, including both glenoids in ten normal volunteers. Shoulders without an osseous fragment at the anteroinferior portion of the glenoid were compared with the contralateral shoulder in the same patient to determine if the glenoid morphology was normal. In shoulders with an osseous fragment, the fragment was evaluated quantitatively and its size was classified as large (>20% of the glenoid fossa), medium (5% to 20%), or small (<5%). Finally, all 100 shoulders were evaluated arthroscopically to confirm the presence of the lesion at the glenoid rim that had been identified with three dimensionally reconstructed computed tomography. RESULTS: Investigation of the normal glenoids revealed no side-to-side differences. Investigation of the affected glenoids revealed an abnormal configuration in ninety shoulders. Fifty glenoids had an osseous fragment. One fragment was large (26.9% of the glenoid fossa), twenty-seven fragments were medium (10.6% of the glenoid fossa, on the average), and twenty-two were small (2.9% of the glenoid fossa, on the average). In the forty shoulders without an osseous fragment, the anteroinferior portion of the glenoid appeared straight on the en face view and it appeared obtuse or slightly rounded, compared with the normally sharp contour of the normal glenoid rim, on the oblique view, suggesting erosion or a mild compression fracture at this site. Arthroscopic investigation revealed a Bankart lesion in ninety-seven of the 100 shoulders and an osseous fragment in forty-five of the fifty shoulders in which an osseous Bankart lesion had been identified with the three dimensionally reconstructed computed tomography. In the shoulders with distinctly abnormal morphology on three-dimensionally reconstructed computed tomography, the arthroscopic appearance of the anteroinferior portion of the glenoid rim was compatible with the appearance demonstrated by the three-dimensionally reconstructed computed tomography. CONCLUSIONS: We introduced a method to evaluate the morphology of the glenoid rim and to quantify the osseous defect in a simple and practical manner with three-dimensionally reconstructed computed tomography with elimination of the humeral head. Fifty percent of the shoulders with recurrent anterior glenohumeral instability had an osseous Bankart lesion; 40% did not have an osseous fragment but demonstrated loss of the normal circular configuration on the en face view and an obtuse contour on the oblique view, suggesting erosion or compression of the glenoid rim. PMID- 12728041 TI - Comparison of arthroscopic findings with magnetic resonance imaging and arthrography in children with glenohumeral deformities secondary to brachial plexus birth palsy. AB - BACKGROUND: Characterization of glenohumeral deformities secondary to brachial plexus birth palsy with plain radiography is difficult because the glenohumeral joint does not completely ossify until puberty. The purpose of this study was to compare the findings on magnetic resonance imaging and arthrography with those on arthroscopy to better understand the roles of these methods in the evaluation of glenohumeral development in this condition. METHODS: Eighty-four children who ranged in age from seven months to thirteen years and six months had glenohumeral arthrography while they were under general anesthesia for operative treatment of an internal rotation contracture. Thirty-six children also received magnetic resonance imaging with use of cartilage-sensitive axial gradient-echo sequences. Thirty-seven children were evaluated arthroscopically. RESULTS: Arthrography showed a concentric glenohumeral joint in thirty-three children, a flat glenoid in eight, a biconcave glenoid in seventeen, and a pseudoglenoid in twenty-six. Thus, 61% (fifty-one) of the eighty-four children with an internal rotation contracture had a substantial deformity. The severity of the contracture was associated with the existence and the type of the deformity (p = 0.001). Magnetic resonance imaging showed greater detail than arthrography did in defining the severity of the deformity in both the glenoid and the humeral head. The thirty seven children who were examined arthroscopically showed a progression from those who had a concentric, conforming joint to those who had a markedly deformed joint with a bifurcated glenoid and a flattened, oval-shaped humeral head that articulated with the posterior aspect of the glenoid. Irregularities and cavitation of the anterior aspect of the glenoid were common. The subscapularis and rotator interval tissue were the primary sites of contracture. CONCLUSIONS: Profound glenohumeral deformities secondary to brachial plexus birth palsy are commonly seen within the first two years of life. The information provided by imaging studies is helpful in defining the natural history of this condition and in determining the success of surgical intervention. PMID- 12728042 TI - Hip arthroplasty for salvage of failed treatment of intertrochanteric hip fractures. AB - BACKGROUND: Failed treatment of an intertrochanteric fracture typically leads to profound functional disability and pain. Treatment with repeated attempts to gain union and to preserve the host femoral head usually is preferred for young patients, but salvage treatment with hip arthroplasty may be considered for selected older patients with poor bone quality, bone loss, or articular cartilage damage. The purpose of the present study was to evaluate the results and complications of hip arthroplasty performed as a salvage procedure after the failed treatment of an intertrochanteric hip fracture. METHODS: Between 1985 and 1997, sixty patients (forty-nine women and eleven men) with a mean age of seventy eight years were treated at our institution with hip arthroplasty after the failed treatment of an intertrochanteric fracture. Thirty-two patients had a total hip arthroplasty with a cemented cup (twenty-four patients) or an uncemented cup (eight patients), twenty-seven had a bipolar hemiarthroplasty, and one had a unipolar hemiarthroplasty. A calcar-replacement design, extended-neck stem, or long-stem implant was used in fifty-one of the sixty hips. RESULTS: Ten patients died within two years (all with the implant intact), and six were lost to follow-up. The remaining forty-four patients were followed for a mean of five years (range, two to fifteen years). At the time of the last follow-up, thirty nine patients had no or mild pain and five had moderate or severe pain; in all of these patients, the pain was in the region of the greater trochanter. Forty patients were able to walk, twenty-six with one-arm support or less. Twelve patients had a total of thirteen medical complications postoperatively. A total of five reoperations were performed: two patients had a revision, one had a rewiring procedure because of trochanteric avulsion, one had late removal of trochanteric hardware, and one had debridement of fat necrosis. One patient had two dislocations, both of which were treated with closed reduction. Kaplan-Meier survivorship analysis with revision of the implant for any reason as the end point revealed a survival rate of 100% at seven years and 87.5% (95% confidence interval, 67.3% to 100%) at ten years. CONCLUSIONS: Hip arthroplasty is an effective salvage procedure after the failed treatment of an intertrochanteric fracture in an older patient. Most patients have good pain relief and functional improvement. Calcar-replacement and long-stem implants often are required. Despite the operative challenges, surprisingly few serious orthopaedic complications were associated with this procedure in the present study. PMID- 12728044 TI - Epinephrine-induced pulmonary edema during arthroscopic knee surgery. A case report. PMID- 12728043 TI - Effect of regional gene therapy with bone morphogenetic protein-2-producing bone marrow cells on spinal fusion in rats. AB - BACKGROUND: Bone morphogenetic proteins (BMPs) are now being used as bone-graft substitutes to enhance spinal fusion. However, the large doses of BMP required to induce a spinal fusion in humans suggests that the delivery of these proteins should be improved. We used ex vivo adenoviral gene transfer to create BMP-2 producing bone marrow cells, and these autologous cells were found to induce a posterolateral fusion of the spine in syngeneic rats. METHODS: Intertransverse spinal arthrodesis (L4 and L5) was attempted in ten groups of Lewis rats with 5 x 10 (6) BMP-2-producing rat bone marrow cells (Ad-BMP-2 cells), created through adenoviral gene transfer with guanidine hydrochloride-extracted demineralized bone matrix as a carrier (Group I); 5 x 10 (6) Ad-BMP-2 cells on a collagen sponge carrier (Group II); 10 micro g of recombinant BMP-2 (rhBMP-2) in a guanidine hydrochloride-extracted demineralized bone matrix carrier (Group III); 10 micro g of rhBMP-2 in a collagen sponge carrier (Group IV); autogenous iliac crest bone-grafting (Group V); 5 x 10 (6) beta-galactosidase-producing rat bone marrow cells, created through adenoviral gene transfer with guanidine hydrochloride-extracted demineralized bone matrix as a carrier (Group VI); decortication of the transverse processes alone (Group VII); 5 x 10 (6) uninfected rat bone marrow cells with a guanidine hydrochloride-extracted demineralized bone matrix carrier (Group VIII); guanidine hydrochloride-extracted demineralized bone matrix only (Group IX); or a collagen sponge alone (Group X). Each specimen underwent plain radiography, manual palpation, and histological analysis. RESULTS: All spines in Groups I and II (BMP-2-producing bone marrow cells) and all spines in Groups III and IV were fused at four weeks postoperatively. In contrast, none of the spines in the other groups had fused at a minimum of eight weeks after implantation. Histological analysis of the specimens revealed that the spines that had received BMP-2-producing bone marrow cells (Groups I and II) were filled with coarse trabecular bone postoperatively, whereas those that had received rhBMP-2 (Groups III and IV) were filled with thin, lace-like trabecular bone. All of the other spines, including those that had been treated with autogenous iliac crest bone-grafting (Group V), produced little or no new bone. CONCLUSION: BMP-2-producing bone marrow cells, created by adenoviral gene transfer, produce sufficient BMP to induce an intertransverse fusion in the rat spine model. PMID- 12728045 TI - Peroneal nerve palsy following acupuncture treatment. A case report. PMID- 12728046 TI - Stent graft treatment of an external iliac artery injury associated with pelvic fracture. A case report. PMID- 12728047 TI - Ankle arthritis. PMID- 12728049 TI - An AOA critical issue. Less invasive procedures in spine surgery. PMID- 12728051 TI - Endoscopic compared with open carpal tunnel release. PMID- 12728052 TI - Gene therapy and the future of orthopaedics. PMID- 12728053 TI - Factor replacement and outcomes in hemophiliacs undergoing total knee arthroplasty. PMID- 12728054 TI - Sequelae of Hylan G-F 20 viscosupplementation of the knee. PMID- 12728055 TI - Etiology of mental status changes in patients after total hip arthroplasty: hypoxemia or paradoxical embolization of fat or bone marrow? PMID- 12728056 TI - Risk of infection after total knee arthroplasty in HIV-positive hemophilic patients. PMID- 12728057 TI - A historical note on anterior cruciate ligament reconstruction. PMID- 12728058 TI - Reviewing the data on survival rates of stems and cups in total hip replacement. PMID- 12728060 TI - Cementless and cemented implants had similar survival in total knee replacement. PMID- 12728062 TI - Conservative treatment reduced pain and was as effective as sutures in hand lacerations. PMID- 12728064 TI - Bed rest and normal daily activity were equivalent for acute low-back pain. PMID- 12728066 TI - What's new in pediatric orthopaedics. PMID- 12728067 TI - Physician knowledge of catch-up regimens and contraindications for childhood immunizations. AB - OBJECTIVES: To determine physician success at designing catch-up regimens for children delayed in immunizations and physician knowledge regarding contraindications to immunization. METHODS: A self-administered survey was completed by pediatricians, general practitioners, and family practitioners in Cook County, Illinois. Surveys included 6 open-ended vignettes describing hypothetical children delayed in immunization for whom participants were asked to design catch-up regimens. Bivariate and multivariate logistic regression were used to determine predictors of correct response. The surveys also inquired about management of scenarios that might be perceived as contraindications to immunize with the Haemophilus influenzae type b or measles-mumps-rubella vaccines. RESULTS: The mean score of correct responses was 1.83 of a possible 6.0. Almost one third of respondents answered all 6 vignettes incorrectly. The proportion of incorrect responses was high for all 6 vignettes (39%-86%), but higher for questions that addressed the immunization of children older than 12 months. Errors in vaccine administration were most commonly attributed to omitted vaccines, with varicella-zoster vaccine and pneumococcal conjugate vaccine omitted most frequently. Pediatricians were >4 times more likely to answer correctly than were family practitioners. Participants in the Vaccines for Children (VFC) program were more than twice as likely to answer correctly than were non-VFC providers. Knowledge of contraindications was inconsistent, particularly for measles-mumps-rubella vaccine. CONCLUSIONS: Childhood vaccine providers have substantial knowledge deficits of recommended immunization schedules and vaccine contraindications that may contribute to missed opportunities to immunize. Pediatricians and participants in the VFC program were more successful at designing catch-up regimens for children with immunization delay. PMID- 12728068 TI - Continuous subcutaneous glucose monitoring improved metabolic control in pediatric patients with type 1 diabetes: a controlled crossover study. AB - OBJECTIVE: To improve metabolic control and prevent complications, both acute and late, we need to adjust treatment on the basis of the blood glucose (BG) profile, as not even the most active BG self-monitoring gives sufficient information. DESIGN: We have used Continuous Glucose Monitoring System (CGMS; Medtronic MiniMed, Northridge, CA) in a controlled crossover study including 27 diabetic patients aged 12.5 +/- 3.3 (mean; standard deviation; range: 5-19) years. All patients were treated with intensive insulin therapy, 14 with multiple injections, and 13 with pumps. The patients were randomized into an open or blind study arm. Both arms wore the CGMS sensor for 3 days every 2 weeks. CGMS profiles were used in the open study arm to adjust insulin therapy at follow-up visits every 6 weeks. Both the patients and the diabetes team were masked to the CGMS profiles in the blinded arm, and insulin therapy adjustments were based solely on 7-point BG profiles performed by the patients. At 3 months the 2 study arms were crossed over. RESULTS: Despite initial problems with a device new to both patients and the diabetes team, hemoglobin A(1)C decreased significantly in the open arm (from 7.70%-7.31%) but not in the blind arm (7.75%-7.65%). A total of 26/27 patients experienced daytime low subcutaneous glucose (<3.0 mmol/L;.8 episodes/day; duration 58 +/- 29 minutes; 5.5% of total time), and 27/27 patients had at least 1 nocturnal episode of low subcutaneous glucose (.4 episodes/night; duration 132 +/- 81 minutes; 10.1% of total time). CONCLUSIONS: Use of CGMS facilitated an improved treatment, and patients received new insight and increased motivation. In this study, we found CGMS to be a useful tool for education and improving metabolic control. PMID- 12728069 TI - Regional brain volumes and their later neurodevelopmental correlates in term and preterm infants. AB - OBJECTIVE: To compare regional brain volumes measured in term and preterm infants, and to correlate regional volumes with measures of neurodevelopmental outcome. METHODS: High-contrast, high-resolution magnetic resonance imaging scans were acquired in 10 preterm and 14 term infants who were scanned near term. The cerebrum was segmented into cortical gray matter, white matter, cerebral ventricles, subcortical gray matter, cerebellum, and brainstem. The cortical gray matter, white matter, and ventricles were further divided into specific anatomic subregions, and the volumes were compared across groups. Measures of cognitive and motor development were acquired between 18 and 20 months of corrected age. Correlations of regional brain volumes with developmental outcome were assessed in the preterm group. RESULTS: Volumes in preterm infants were reduced in parieto occipital gray matter and increased in the midbody, occipital horn, and temporal horns of the lateral ventricles. Gray matter volumes were also less prominently reduced in the sensorimotor and inferior occipital cortices. Normal lateralization of white matter volumes were altered in the parieto-occipital region in the preterm infants, who had significantly larger left-sided and smaller right-sided structures. White matter volumes in the sensorimotor and midtemporal regions correlated strongly with measures of neurodevelopmental outcome. CONCLUSIONS: These findings of reduced volumes in sensorimotor and parieto-occipital regions in preterm infants, and the prospective correlations of regional volumes with cognitive outcome, confirm and extend findings previously reported in a cross-sectional study of 8-year-old prematurely born children. The data suggest that regional brain volumes near term are a promising marker for predicting disturbances of cognitive outcome in preterm infants. Further prospective, longitudinal studies of neonatal brain volumes and developmental indices into later childhood are required to confirm the utility of regional brain volumes as predictors of longer term outcome. PMID- 12728070 TI - Ten-year prospective study of public health problems associated with early drinking. AB - OBJECTIVE: To compare early nondrinkers, experimenters, and drinkers on the prevalence of problem behaviors at grades 7 and 12 and at age 23 (N = 6338, 4265, and 3369, respectively). METHODS: Results are based on longitudinal self-report data from individuals who were originally recruited from 30 California and Oregon schools at grade 7 (1985) and assessed again at grade 12 (1990) and at age 23 (1995). Logistic regression was used to develop weighted estimates of the prevalence of academic difficulties, employment problems, substance use, and delinquent and violent behaviors within the 3 drinking status groups at grades 7, 12, and/or at age 23. Huber variance estimates, which adjust for weighting and clustering of observations, were used to assess the statistical significance of differences across groups. RESULTS: Early drinkers and experimenters were more likely than nondrinkers to report academic problems, substance use, and delinquent behavior in both middle school and high school. By young adulthood, early alcohol use was associated with employment problems, other substance abuse, and criminal and violent behavior. CONCLUSIONS: Early drinkers do not necessarily mature out of a problematic lifestyle as young adults. Interventions for these high-risk youth should start early and address their other public health problems, particularly their tendency to smoke and use other illicit drugs. PMID- 12728071 TI - Health services use by children of migratory agricultural workers: exploring the role of need for care. AB - OBJECTIVE: For migrant children: 1). to assess the determinants of health services use among users and nonusers of health services; 2). to evaluate the association between health status and health services use, while controlling for potential confounders. DESIGN: A cross-sectional household survey using multistage, partially random sampling to identify migrant families in eastern North Carolina. PARTICIPANTS: Adult caretakers of 1 randomly selected child under 13 years old. RESULTS: Forty-four percent of children (N = 300) visited a doctor in the preceding 3 months. Those visiting a doctor disproportionately reported having less than very good health (29% vs 10%), insurance (46% vs 11%), interpreters (45% vs 27%), a family member receiving Special Supplemental Nutrition Program for Women, Infants, and Children (50% vs 16%), and a legal caretaker (30% vs 18%). Compared with those without a doctor visit, a larger proportion of children visiting a doctor were 6 years or younger in age (71% vs 35%), born in the United States (51% vs 15%), female (64% vs 45%), and had not moved in 6 or more months (19% vs 10%). Controlling for enabling resource and sociodemographic confounders, children with less than very good health were 2.4 times more likely than those in very good health to have visited a doctor (95% confidence interval [1.1-5.2]). CONCLUSIONS: Migrant children using health services are distinct from nonusers with regards to sociodemographic factors, enabling resources, and need for care. Health services use is associated with less than very good perceived health, despite resource barriers and sociodemographic disadvantages. More efforts are needed to improve access to health care for migrant children. PMID- 12728072 TI - Serious bacterial infections in febrile infants younger than 90 days of age: the importance of ampicillin-resistant pathogens. AB - BACKGROUND: Intrapartum antibiotic prophylaxis against group B Streptococcus (GBS) has reduced the occurrence of serious bacterial infections (SBI) in young infants caused by GBS. Recommendations for initial antibiotic therapy for the febrile infant 1 to 90 days old were developed when infections with GBS were common and antibiotic resistance was rare. OBJECTIVE: To document the pathogens responsible for SBI in recent years in febrile infants 1 to 90 days old and the antibiotic susceptibility of these organisms. METHODS: The results of bacterial cultures from infants 1 to 90 days old evaluated for fever at Primary Children's Medical Center in Salt Lake City, Utah, between July 1999 and April 2002 were analyzed. Antibiotic susceptibility profiles were collected and patient records were reviewed to determine if initial antibiotic therapy was changed following the identification of the organism. RESULTS: Of 1298 febrile infants enrolled from the Primary Children's Medical Center emergency department, 105 (8%) had SBI. The mean age of the infants with SBI was 39 days (range 2-82 days) and 2 (2%) were <7 days. SBI included urinary tract infection (UTI; 67%), bacteremia (16%), bacteremia and UTI (6%), bacteremia and meningitis (5%), meningitis (2%), abscess (2%), meningitis and UTI (1%), and meningitis and gastroenteritis (1%). Eighty-three (79%) of 105 episodes of SBI were caused by Gram-negative bacteria, including 92% of UTI, 54% of bacteremia, and 44% of meningitis cases. The most common pathogen was Escherichia coli (61%). Other Gram-negative pathogens were responsible for 19% of SBI. Staphylococcus aureus was the most common Gram positive pathogen, causing 8% of SBI. GBS accounted for 6% of SBI. Of the 105 pathogens, 56 (53%) were resistant to ampicillin. Of the pathogens causing meningitis, UTI, and bacteremia, 78%, 53%, and 50%, respectively, were resistant to ampicillin. Antibiotic therapy was changed in 54% of cases of SBI following identification of the organism. CONCLUSIONS: In Utah, ampicillin-resistant Gram negative bacteria are the most common cause of SBI in febrile infants <90 days old. This finding impacts antibiotic selection, especially in cases of meningitis. Local surveillance of pathogens and antibiotic susceptibility patterns is critical to determine appropriate antibiotic therapy. PMID- 12728073 TI - Psychopathology and social competencies of adolescents who were extremely low birth weight. AB - BACKGROUND: We have previously shown that infants who were extremely low birth weight (ELBW) are particularly vulnerable to problems related to inattention and hyperactivity at school age. It is not known whether these problems persist to adolescence. OBJECTIVE: To explore and compare the levels of psychopathology in a regional cohort of ELBW infants and sociodemographically matched term controls as reported by teens and their parents. DESIGN/METHODS: Cross-sectional cohort study/geographically defined region. PARTICIPANTS: TEENS: ELBW 141/169 (83%) and control 122/145 (84%), aged 12 to 16 years. PARENTS: ELBW 143/169 (85%) and control 123/145 (85%). Both cohorts and their parents completed the Ontario Child Health Study-Revised questionnaire with 6 behavioral subsca1es: conduct disorder, oppositional defiant disorder, attention-deficit/hyperactivity disorder (ADHD), overanxious, separation anxiety, and depression. RESULTS: By teen self-report, there were no significant differences between ELBW and control teens on any of the 6 subscale scores. However, parents of ELBW teens reported significantly higher scores than parents of control for depression and ADHD based on 2-way analysis of variance (group x gender). Comparison within teen/parent dyads showed that both cohorts of teens reported significantly higher scores than their parents. Multivariable analyses on behavioral subscale scores demonstrated a number of variables that were significant predictors by parent report: group (ELBW vs control), gender, family function, developmental quotient, maternal mood, and socioeconomic status; no predictors were significant by teen report. There were no statistically significant interaction effects for any of the models. These models explained a greater percent of the variance in behavioral scores for parents than for teens (12.5%-22.0% vs 3.4%-8.2%). Results were similar when teens with neurosensory impairment were excluded. CONCLUSIONS: This study is unique in the inclusion of both parent and teen self-report of behaviors. Significant differences were apparent only by parent report for ADHD and depression among ELBW teens. Significant predictors of behavioral scores for parents include group, gender, family function, developmental quotient, maternal mood, and socioeconomic status. PMID- 12728074 TI - Resident documentation discrepancies in a neonatal intensive care unit. AB - CONTEXT: Medical errors are common and potentially dangerous. Little is known about the role of documentation errors. OBJECTIVE: To determine the proportion of resident physician progress notes that contained discrepancies, and to identify predictors of such discrepancies. DESIGN/METHODS: We conducted a retrospective cross-sectional chart review of resident physician progress notes over 40 random days in a 4-month period in a neonatal intensive care unit. Using predetermined criteria, we compared resident documentation of patient weights, medications, and vascular lines to other sources of information and recorded the numbers of documentation discrepancies. RESULTS: There were discrepancies in 209 (61.7%) notes with respect to weight, vascular lines, or medications. Discrepancies occurred in the documentation of medications in 103 (27.7%) progress notes, of vascular lines in 119 (33.9%) progress notes, and of weights in 45 (13.3%) progress notes. Notes both omitted information regarding medications (18.2%) and vascular lines (13.9%) and documented inaccurate information regarding medications (18.6%) and vascular lines (30.1%). Patients with more medications or vascular lines, and with longer lengths of stay, were significantly more likely to have higher rates of documentation errors. CONCLUSIONS: Daily progress notes written by resident physicians in the neonatal intensive care unit often contain inaccurate, or omit pertinent, information. Alternative means or methods of documentation are warranted. PMID- 12728075 TI - Herbal therapy use in a pediatric emergency department population: expect the unexpected. AB - BACKGROUND: In recent years investigators have reported widespread use of alternative medicine. Some herbal therapies have potentially harmful side effects as well as adverse interactions with medications. Data are lacking on the use in children and caregiver understanding of these products. OBJECTIVES: To determine the reported use of herbal products among a pediatric emergency department population and to evaluate the caregivers' understanding and source of information concerning these products. DESIGN/METHODS: A convenience sampling of pediatric emergency department patients and their caregivers occurred during a 3 month period in 2001. The interview consisted of 18 questions regarding the types of non-Food and Drug Administration-regulated herbal products and home remedies used, general product knowledge and sources of information used by the child's caregiver (including discussions with their child's primary physician). RESULTS: One hundred forty-two (93%) of 153 families approached participated in the study. The mean patient age was 5.3 years (range: 3 weeks-18 years). Forty-five percent of caregivers reported giving their child an herbal product, and 88% of these caregivers had at least 1 year of college education. Of the children receiving these therapies, 53% had been given 1 type and 27% were given 3 or more in the past year. The most common therapies reportedly used were aloe plant/juice (44%), echinacea (33%), and sweet oil (25%). The most dangerous potential herbal and prescription medication combination reported was ephedra and albuterol in an adolescent with asthma. The most unusual products reportedly used included turpentine, pine needles, and cowchips. Of all people interviewed, 77% did not believe or were uncertain if herbal products had any side effects and only 27% could name a potential side effect. Sixty-six percent were unsure or thought that herbal products did not interact with other medications and only 2 people correctly named a drug interaction. Of the people who used these therapies, 80% reported either friends or relatives as their primary source of information. Only 45% of those giving their children herbal products report discussing the use with their child's primary health care provider. CONCLUSION: Herbal and home therapies are commonly used in this pediatric population. An unexpectedly wide variety of products were reportedly given to this patient population. Caregivers reported limited knowledge regarding potential adverse medication interactions and side effects. Limited discussions with the child's primary health care provider were reported. It is therefore important for health care providers to have knowledge about herbal medications, to inquire about their use and to educate families about the risk/benefit as well as potential interactions these products may have with over-the-counter and prescription medications. PMID- 12728076 TI - Extrauterine growth restriction remains a serious problem in prematurely born neonates. AB - BACKGROUND: Poor growth is a common problem in premature neonates and may be associated with neurodevelopmental delay. OBJECTIVE: To evaluate the incidence of extrauterine growth restriction (growth values < or =10th percentile of intrauterine growth expectation based on estimated postmenstrual age in premature (23-34 weeks' estimated gestational age) neonates at the time they are discharged from the hospital. DESIGN/METHODS: Using a database formed from a computer assisted tool that generates clinical progress notes and discharge summaries on neonatal intensive care unit admissions, we reviewed data on neonates discharged from 124 neonatal intensive care units between January 1, 1997, and December 31, 2000. We evaluated neonates who were born between 23 and 34 weeks' estimated gestational age without congenital anomalies and who were cared for at and discharged from the same hospital. For each patient, we compared the discharge growth values to the expected values based on our intrauterine growth data and postmenstrual age on the day of discharge. We defined extrauterine growth restriction as having a measured growth value (weight, length or head circumference) that was < or =10th percentile of the predicted value. In each specific birth weight and estimated gestational age group, we counted the number of neonates < or =10th percentile for each growth parameter and calculated the percentage of patients who had values < or =10th percentile at discharge. Using logistic regression, we evaluated the factors associated with extrauterine growth restriction for weight, length, and head circumference. RESULTS: Our sample included 24 371 premature neonates. Data on discharge weight, length, and head circumference was available on 23 970, 17 203, and 20 885 neonates, respectively. The incidence of extrauterine growth restriction was common (28%, 34%, and 16% for weight, length, and head circumference, respectively). For each growth parameter, the incidence of extrauterine growth restriction increased with decreasing estimated gestational age and birth weight. Factors independently associated with extrauterine growth restriction were male gender, need for assisted ventilation on day 1 of life, a history of necrotizing enterocolitis, need for respiratory support at 28 days of age, and exposure to steroids during the hospital course. CONCLUSIONS: Extrauterine growth restriction remains a serious problem in premature neonates especially for neonates who are small, immature, and critically ill. PMID- 12728078 TI - Nutritional state and growth and functional maturation of the brain in extremely low birth weight infants. AB - OBJECTIVE: It is well-known that an undernutritional status influences central nervous system development in the fetal and early neonatal period. On the other hand, the maturational delay of the central nervous system is reflected as dysmature pattern (DMP) in the neonatal background electroencephalograph (EEG). Therefore, we hypothesized that the postnatal nutritional status influenced electrophysiologic maturation in extremely low birth weight infants (ELBWIs). METHODS: ELBWIs between 24 and 27 weeks of gestational age who were admitted to Ogaki Municipal Hospital NICU from April 1997 to December 2000 were considered eligible. From the condition of enteral feeding, infants were divided into 2 groups: 1). normal nutritional group (group N), where enteral feeding had been established (100 mL/kg/d) by 3 weeks after birth; 2). undernutritional group (group U), where enteral feeding had not been established by 3 weeks after birth or was discontinued because of clinical problems. Weekly average body weight and head circumference gains were evaluated as nutritional status. EEG records were performed every 2 to 4 weeks until postnatal 15 weeks of age. DMP was defined as the appearance of immature EEG patterns for postconceptional age. RESULTS: Twenty one infants had serial EEG recordings; 11 infants belonged to group N and 10 infants to group U. Gestational age, birth weight, and head circumference at birth were not different between the 2 groups. The body weight of group N was significantly heavier than that of group U after 5 postnatal weeks. Similarly, the head circumference of group N was larger than that of group U after 6 weeks of postnatal age. Nine infants demonstrated DMPs. One infant belonged to group N and 8 to group U. DMPs were significantly more frequently found in group U than group N (80% vs 9%). In 6 of the 9 cases, the DMPs lasted until 38 to 40 weeks of postconceptional age. Five of the 6 infants with persistent DMPs suffered from severe undernutritional conditions. The other, who belonged to group N, was treated with corticosteroid for chronic lung disease. In 3 cases, DMPs were observed transiently and their undernutritional status was not so severe. CONCLUSIONS: Our study indicated that a postnatal undernutritional condition was associated with DMPs in ELBWIs. Undernutritional status may affect electrophysiologic maturation. PMID- 12728079 TI - Missed opportunities for sexually transmitted diseases, human immunodeficiency virus, and pregnancy prevention services during adolescent health supervision visits. AB - OBJECTIVE: To describe prevention counseling on pregnancy and sexually transmitted diseases (STDs), including human immunodeficiency virus (HIV), received by sexually experienced youth in the primary care setting and to test associations between recent sexual risk behaviors and preventive counseling. METHODS: Using data from the 1999 Youth Risk Behavior Surveillance survey, a nationally representative survey (N = 15 349) of high school students, we analyzed responses to questions about sexual experience, time since last preventive health care visit, and discussion of STD, HIV, or pregnancy prevention with a doctor or nurse during their last preventive health care visit. Logistic regression was used to test associations; students' demographic characteristics were controlled. RESULTS: More than half of the US high school students surveyed reported a preventive health care visit in the 12 months preceding the survey: 60.4% (95% confidence interval [CI]: 57.2%-63.6%) of female students and 57.5% (95% CI: 53.9%-61.1%) of male students. For female students, sexual experience was positively associated with a preventive health care visit (odds ratio [OR]: 1.3; 95% CI: 1.1-1.6), but for male students, sexual experience had a negative effect (OR: 0.8; 95% CI: 0.7-0.9). Of the students who reported a preventive health care visit in the 12 months preceding the survey, 42.8% (95% CI: 38.6% 47.1%) of female students and 26.4% (95% CI: 22.7%-30.2%) of male students reported having discussed STD, HIV, or pregnancy prevention at those visits. Sexual experience was associated with a higher likelihood of engaging in a dialogue about sexual health once a student entered the health care system: female students (OR: 3.8; 95% CI: 3.0-4.9) and male students (OR: 1.9; 95% CI: 1.3-2.7). CONCLUSION: Primary care providers miss opportunities to provide STD, HIV, and pregnancy prevention counseling to high-risk youth. PMID- 12728080 TI - The effect of zinc supplementation on linear growth, body composition, and growth factors in preterm infants. AB - OBJECTIVE: The aim of our study was to evaluate the effect of zinc supplementation on linear growth, body composition, and growth factors in premature infants. DESIGN: Thirty-six preterm infants (gestational age: 32.0 +/- 2.1 weeks, birth weight: 1704 +/- 364 g) participated in a longitudinal double blind, randomized clinical trial. They were randomly allocated either to the supplemental (S) group fed with a standard term formula supplemented with zinc (final content 10 mg/L) and a small quantity of copper (final content 0.6 mg/L), or to the placebo group fed with the same formula without supplementation (final content of zinc: 5 mg/L and copper: 0.4 mg/L), from 36 weeks postconceptional age until 6 months corrected postnatal age. At each evaluation, anthropometric variables and bioelectrical impedance were measured, a 3-day dietary record was collected, and a blood sample was taken. We analyzed serum levels of total alkaline phosphatase, skeletal alkaline phosphatase (sALP), insulin growth factor (IGF)-I, IGF binding protein-3, IGF binding protein-1, zinc and copper, and the concentrations of zinc in erythrocytes. RESULTS: The S group had significantly higher zinc levels in serum and erythrocytes and lower serum copper levels with respect to the placebo group. We found that the S group had a greater linear growth (from baseline to 3 months corrected age: Delta score deviation standard length: 1.32 +/-.8 vs.38 +/-.8). The increase in total body water and in serum levels of sALP was also significantly higher in the S group (total body water: 3 months; corrected age: 3.8 +/-.5 vs 3.5 +/-.4 kg, 6 months; corrected age: 4.5 +/ .5 vs 4.2 +/-.4 kg; sALP: 3 months; corrected age: 140.2 +/- 28.7 vs 118.7 +/- 18.8 micro g/L). CONCLUSIONS: Zinc supplementation has a positive effect on linear growth in premature infants. PMID- 12728081 TI - Growth deficits and attention-deficit/hyperactivity disorder revisited: impact of gender, development, and treatment. AB - OBJECTIVE: Although the relationship between putative growth deficits and attention-deficit/hyperactivity disorder (ADHD) has been examined in boys, this issue has not been evaluated in girls. METHODS: Height and weight were examined in 124 female ADHD children and 116 female controls using age and parental height corrections, attending to issues of pubertal stage and treatment. Also, we examined the interaction between ADHD status and gender on growth outcomes using data from 124 ADHD and 109 control males. RESULTS: The ADHD-growth association was not moderated by gender. No deficits in age-adjusted height or age and height adjusted weight were detected in ADHD girls. Also, we found no association between growth measurements and psychotropic treatment, malnutrition, short stature, pubertal development, family history of ADHD, or psychiatric comorbidity, except for major depression: ADHD girls with major depression were on average 7.6 kg heavier than ADHD girls without depression, adjusting for age and height. CONCLUSIONS: No growth deficits appear to be associated with ADHD or its treatment in females. These findings add to a growing literature supporting the notion that stimulant treatment does not have an adverse impact on ADHD children's growth and development. PMID- 12728083 TI - The ecology of medical care for children in the United States. AB - BACKGROUND: Medical care ecology has previously been investigated for adults, but no similar exploration has been made specifically for children. OBJECTIVE: To describe proportions of children receiving care in 6 types of health care setting on a monthly basis and to identify characteristics associated with receipt of care in these settings. METHODS: Nationally representative data from the 1996 Medical Expenditure Panel Survey were used to estimate the number of children per 1000 per month who received care at least once in each type of setting. Multivariate analyses assessed associations between receiving care in various settings and children's sociodemographic factors (age, sex, ethnicity, poverty, parent's education, urban or rural residence, insurance status, and whether or not the child had a usual source of care). RESULTS: Of 1000 children aged 0 to 17 years, on average each month 167 visited a physician's office, 82 a dentist's office, 13 an emergency department, and 8 a hospital-based outpatient clinic. Three were hospitalized and 2 received professional health care in their home. Younger age was associated with increased proportions of children receiving care in all health care settings except dentists' clinics. Poverty, lack of health insurance, black race, and Hispanic ethnicity were associated with decreased receipt of care in physicians' and dentists' offices. Only age (<2 years and 13 17 years) and poverty status were associated with hospitalization (P <.05 for each). Rural residence was not associated with any significant variation in proportions of children receiving care in any setting. Having a usual source of care was associated with increased receipt of care in all settings except hospitals. CONCLUSIONS: The ecology of children's medical care is similar to that of adults in the United States in that a substantial proportion of children receive health care each month, mostly in community-based, outpatient settings. In all settings except emergency departments, receipt of care varies significantly by children's age, race, ethnicity, income, health insurance status, and whether they have a usual source of care. PMID- 12728082 TI - Reduced bone mineralization in infants fed palm olein-containing formula: a randomized, double-blinded, prospective trial. AB - OBJECTIVE: Palm and palm olein (PO) oils are used in some infant formula fat blends to match the fatty acid profile of human milk, but their presence has been shown to lower calcium and fat absorption. We aimed to determine if the reported differences in calcium absorption could affect skeletal development by comparing bone mineral accretion in healthy term infants fed a milk-based formula with (PMF) or without PO. METHODS: Whole body bone mineralization was evaluated in healthy term infants fed 1 of 2 coded, commercially available, ready-to-feed infant formulas in a randomized, double-blind, parallel study. Subjects were fed either 1). PMF formula (Enfamil with iron; Mead Johnson Division of Bristol Myers, Evansville, IN; N = 63) containing PO/coconut/soy/high-oleic sunflower oils (45/20/20/15% oil); or 2). milk-based formula without PO (Similac with iron; Ross Products Division Abbott Laboratories, Columbus, OH; N = 65), containing high-oleic safflower/coconut/soy oils (40/30/30% oil) from enrollment by 2 weeks after birth until 6 months. Anthropometrics and formula intake were determined monthly; total body bone mineral content (BMC) and bone mineral density (BMD) were measured at baseline, 3, and 6 months of age using dual energy x-ray absorptiometry. RESULTS: Intent-to-treat analyses indicated no significant differences between feeding groups in weight, length, head circumference, or formula intake throughout the study. BMC and BMD were not different at baseline but repeated measures analyses show that infants fed PMF had significantly lower BMC and BMD at 3 and 6 months. CONCLUSIONS: Healthy term infants fed a formula containing PO as the predominant oil in the fat blend had significantly lower BMC and BMD than those fed a formula without PO. The inclusion of PO in infant formula at levels needed to provide a fatty acid profile similar to that of human milk leads to lower bone mineralization. PMID- 12728084 TI - Height and weight development during four years of therapy with cyclical intravenous pamidronate in children and adolescents with osteogenesis imperfecta types I, III, and IV. AB - OBJECTIVES: Treatment with pamidronate improves the clinical course in children with osteogenesis imperfecta (OI), but theoretically might affect longitudinal growth. In this study we analyzed growth during cyclical intravenous pamidronate treatment in children and adolescents (age.04-15.6 years at baseline) with moderate to severe forms of OI types I, III, and IV. METHODS: The effect of 1 year of pamidronate treatment on height and weight was analyzed in 116 patients (OI-I, N = 29; OI-III, N = 42; OI-IV, N = 45). The results of 4 years of therapy were evaluated in 41 children (OI-I, N = 12; OI-III, N = 14; OI-IV, N = 15). RESULTS: Baseline height was low for age in all OI types. After 1 year of pamidronate therapy, height z scores had increased significantly in OI-III (by 0.3 +/- 0.8, mean +/- standard deviation; P =.04) and did not change in OI-I and OI-IV. Weight z scores increased significantly in OI-I (by 0.2 +/- 0.4, P =.01). After 4 years of pamidronate therapy, mean height z scores increased significantly in OI-IV (by 0.41 +/- 0.71, P =.04), whereas nonsignificant trends to increase were found for OI-I and OI-III. When height was expressed as a percentage of the result expected for untreated patients with the same OI type, long-term pamidronate therapy was associated with a significant height gain in all 3 OI types (P <.001). Eight patients who reached final height after 3.0 +/- 1.0 years of treatment were taller on average than expected for untreated patients (P =.04). CONCLUSIONS: Four years of cyclical intravenous pamidronate treatment led to a significant height gain in moderately to severely affected OI patients. PMID- 12728085 TI - Factors influencing parental consent in pediatric clinical research. AB - BACKGROUND: Although previous studies have looked at the motivation behind enrollment in clinical trials of adults and healthy subjects, little is known about the factors influencing parental consent in pediatric clinical studies where the subjects themselves do not provide consent. OBJECTIVE: This study was conducted to define a group of factors that impact parental consent in the pediatric clinical trial. This may allow investigators to identify a subset of specific appealing aspects of clinical trials that will promote participation while also bringing ethical issues to light that may require further consideration. METHODS: Forty-four parents or guardians of children currently participating in clinical asthma research completed questionnaires from July 1999 to September 1999 pertaining to motives for allowing their child to participate in clinical trials. The responses were then rated on a Likert numerical scale. RESULTS: The most important motive for parents is learning more about their child's illness. Next important was the motive of helping medical knowledge. Availability of free medication was negatively correlated with family income. CONCLUSIONS: Although altruistic motives are present in pediatric asthma research, most parents/guardians gave consent for their child to learn more about their child's asthma. Access to free medication was more important in families with lower incomes than in families with higher incomes. PMID- 12728087 TI - Who gets diagnosed with asthma? Frequent wheeze among adolescents with and without a diagnosis of asthma. AB - OBJECTIVE: 1). To describe the factors associated with not receiving an asthma diagnosis among children with frequent wheezing symptoms and 2). to determine risk factors for frequent wheezing in the population. METHODS: The North Carolina School Asthma Survey provided self-reported questionnaire data on respiratory health from 122 829 children ages 12 to 18 years enrolled in 499 public middle schools in North Carolina during the 1999-2000 school year. Questions from the International Survey of Allergies and Asthma in Childhood were used to estimate the prevalence of asthma and wheezing-related illness and associated factors. RESULTS: Factors independently associated with undiagnosed frequent wheezing versus asymptomatic children included female gender (odds ratio [OR]: 1.45; 95% confidence interval [CI]: 1.35-1.54), current smoking (OR: 2.60; 95% CI: 2.43 2.79), exposure to household smoke (OR: 1.59; 95% CI: 1.50-1.70), low socioeconomic status (OR: 1.52; 95% CI: 1.42-1.63), and African American (OR: 1.25; 95% CI: 1.15-1.34), Native American (OR: 1.35; 95% CI: 1.11-1.62), and Mexican American (OR: 1.32; 95% CI: 1.17-1.48) race/ethnicity. Urban residence showed a weak negative association (OR: 0.91; 95% CI: 0.85-0.96). A similar pattern of results was observed for analyses comparing odds of undiagnosed frequent wheeze versus diagnosed asthmatics. Report of allergies was less likely in frequent wheezers (70%) compared with diagnosed asthmatics (86%), but much higher than in asymptomatic children (36%). Thirty-three percent of children with undiagnosed frequent wheezing reported 1 or more physician visits in the last year for wheezing or breathing problems compared with 71% of children with diagnosed asthma, and 4% in asymptomatic children. The prevalence of any inhaler use in the past 12 months was 12% for undiagnosed frequent wheezers versus 78% for diagnosed asthmatics. The proportion of undiagnosed frequent wheezers with fair or poor self-rated health (23%) was slightly higher than diagnosed asthmatics (20%) and much higher than asymptomatic children (4%). CONCLUSIONS: In one of the largest adolescent asthma surveys ever reported in the United States, undiagnosed frequent wheezing was independently associated with female gender, current smoking, exposure to household smoke, low socioeconomic status, allergies, and African American, Native American, and Mexican American race/ethnicity. Children with undiagnosed frequent wheezing were not receiving adequate health care for their asthma-like illness. Clinicians who treat adolescents should consider asking adolescents specifically about wheezing. This information may assist primary care physicians in identifying children with undiagnosed asthma in need of treatment. PMID- 12728086 TI - Immunogenicity of a three-component acellular pertussis vaccine administered at birth. AB - OBJECTIVE: To evaluate within the first 6 months of birth the immunogenicity of a 3-component acellular pertussis (aP) vaccine containing filamentous hemagglutinin (FHA), pertactine (PRN), and genetically detoxified pertussis toxin (PT) in infants who received a dose of vaccine at birth, in addition to the recommended schedule administered at 3, 5, and 11 months. Furthermore, we investigated the influence of maternal antibodies on aP vaccine response. METHODS: We used enzyme linked immunosorbent assay to evaluate immunoglobulin G antibody levels in 45 infants immunized at birth and at 3, 5, and 11 months (group 1) and in 46 infants immunized at the ages of 3, 5, and 11 months (group 2). All mothers were also tested at delivery. RESULTS: At the age of 5 months the geometric mean titer of anti-PT, anti-FHA, and anti-PRN was significantly greater in group 1 (who had received 2 doses) than in group 2 (1 dose). At 6 months geometric mean titers were significantly higher in group 1 than in group 2 for anti-PRN and anti-FHA, whereas no significant differences were observed for anti-PT. CONCLUSIONS: Immunization at birth may be important for an earlier prevention of the pertussis disease in infants under 6 months, especially in Italy, where the recommended ages for aP vaccine administration are 3, 5, and 11 months. PMID- 12728088 TI - Treatment for congenital hypothyroidism: thyroxine alone or thyroxine plus triiodothyronine? AB - OBJECTIVE: To compare the effects of therapy with thyroxine (T4) plus triiodothyronine (T3) versus T4 alone from the first days of life in screened congenital hypothyroid (CH) infants. METHODS: We examined 14 CH infants diagnosed by neonatal screening and a group of control infants. CH patients were divided randomly into 2 groups, 1 treated with T4 alone (group 1) and the other treated with T4 plus T3 (liothyronine; group 2). In all patients electrocardiography and thyroid hormone evaluations were performed before and 15 and 30 days and 3, 6, and 12 months after the beginning of therapy. Psychological tests were also performed at 6 and 12 months of age in CH patients and in other matched controls. RESULTS: After 15 days of treatment, serum thyrotropin (TSH) levels become normal in 5 of 7 cases of group 1 (median TSH level 10.7 micro U/ml) and in 1 of 7 cases of group 2 (median TSH level 72.5 micro U/ml). At the same period, serum-free thyroid hormone levels were within the normal range in both groups, but free T4 values were significantly higher in group 1 than in group 2 and in controls. At the subsequent examinations, free T4 values were within the upper normal limit in group 1, whereas they remained within the normal range in group 2. No clinical or electrocardiographic signs of heart disease were found in any of the patients. The psychometric quotient in CH infants was significantly lower than in controls, but similar in patients of group 1 and group 2. CONCLUSIONS: The combined treatment with T4 plus T3 seems not to show significant advantages, at least in our experimental conditions, compared with the traditional treatment with T4 alone in early treated CH infants. A further longer and more extensive follow-up is mandatory. PMID- 12728089 TI - Antibiotic treatment of acute otorrhea through tympanostomy tube: randomized double-blind placebo-controlled study with daily follow-up. AB - OBJECTIVE: The role of routine antimicrobial treatment of acute middle-ear infections is under debate, because the efficacy of antimicrobials in the resolution of middle-ear fluid has not been unambiguously proven. Acute tube otorrhea is regarded as evidence of acute otitis media, and for methodologic reasons it was chosen to provide objectivity for diagnostics and outcome assessment. The objective of this study was to assess whether amoxicillin clavulanate accelerates the resolution of acute tube otorrhea. DESIGN AND SETTING: Randomized, double-blind, placebo-controlled study in outpatient setting. PATIENTS: Volunteer sample of basically healthy 6- to 72-month-old children with a tympanostomy tube. Eligibility required having acute tube otorrhea of <48 hours' of duration and no prior treatment within the last 2 weeks. The mean age of the participants was 25 months; they had a history of 3 episodes of acute otitis media (median), and 99% had manifestations of a concomitant respiratory infection. Of 79 randomized patients, 7 were withdrawn because of adverse events; 66 patients completed the study. INTERVENTIONS: Amoxicillin-clavulanate (N = 34; 45 mg/kg/d) or matching placebo (N = 32) for 7 days and daily suction of middle-ear fluid through tympanostomy tube. MAIN OUTCOME MEASURES: Duration of acute tube otorrhea and duration of bacterial growth in middle-ear fluid. RESULTS: The median duration of tube otorrhea was significantly shorter in amoxicillin-clavulanate than in the placebo group (3 vs 8 days). At the end of the 7-day medication period, tube otorrhea was resolved in 28 of 34 children receiving amoxicillin-clavulanate compared with 13 of 32 children on placebo (treatment-control difference 41%; 95% confidence interval, 20%-63%; number needed to treat, 2.4). The median duration of bacterial growth in middle-ear fluid was shorter in amoxicillin-clavulanate than in the placebo group (1 vs 8 days). CONCLUSIONS: Oral antibiotic treatment significantly accelerates the resolution of acute tube otorrhea by reducing bacterial growth in middle-ear fluid. PMID- 12728090 TI - Utilization of pediatric hospitals in New York State. AB - BACKGROUND: Little published information is available regarding the proportion of hospitals that offer pediatric services at specified levels, or the actual utilization of such services. OBJECTIVE: To determine if an existing national accreditation process created for educational purposes identifies hospitals with a distinct utilization pattern, in a sample of pediatric inpatient activity from one state. METHODS: Hospitals in New York State were classified as "pediatric" hospitals according to their sponsorship or major participation with an accredited pediatric residence program. Institutions not affiliated with pediatric residencies were considered to be the "other" hospitals. Data on hospital discharges and interhospital transfers were studied for children from 0 to 14 years old, excluding neonatal Diagnosis-Related Groups. Data were obtained from the New York Statewide Planning and Research Cooperative System for 1996, 1997, and 2000. RESULTS: Sixteen percent of hospitals were considered to be "pediatric" facilities (42 of 257) by study criteria. Annual pediatric inpatient activity per "pediatric" hospital significantly exceeded that in "other" institutions, including hospitalizations (2249 +/- 1284 vs 258 +/- 348), number of interhospital transfers received (153 +/- 88 vs 18 +/- 26), and number of hospitals referring interhospital transfers to each receiving hospital (36 +/- 17 vs 5 +/- 7; mean +/- standard deviation; for each comparison). Statewide, "pediatric" hospitals served two thirds of all hospitalized children and 70% of all children's hospital days. "Pediatric" hospitals cared for 26.5 patients per hospital day, compared with only 2.2 per day at "other" institutions. Although statewide hospitalizations of children fell by 14.3% the proportion of all pediatric hospitalizations served by "pediatric" hospitals increased by 3.8% from 1996 to 2000. CONCLUSIONS: Although intended for educational purposes, hospital affiliation with an accredited pediatric residency program identifies a subgroup of facilities with a distinct utilization pattern, which provides care for the majority of pediatric inpatients, and serves as a resource to other hospitals. PMID- 12728091 TI - Autosomal recessive polycystic kidney disease: the clinical experience in North America. AB - OBJECTIVE: We designed a longitudinal clinical database for autosomal recessive polycystic kidney disease (ARPKD), recruited patients from pediatric nephrology centers in the United States and Canada, and examined their clinical morbidities and survival characteristics. We initially targeted enrollment to children who were born and diagnosed after January 1, 1990, so as to capture a cohort that is representative of ARPKD patients born in the last decade. When a significant number of older ARPKD patients were also referred, we extended our database to include all patients who met our inclusion criteria, thereby allowing direct comparisons between a long-term survivor subset and a cohort that included both neonatal survivors and nonsurvivors. DESIGN: Patient entry into our database required either compatible histopathology or ultrasonographic evidence of enlarged, echogenic kidneys and the presence of at least 1 of the following additional criteria: a) biopsy-proven ARPKD in a sibling; b) biliary fibrosis based on either clinical or histopathologic evidence; c) no sonographic evidence of renal cysts in the parents (parents must be >30 years of age); or d) parental consanguinity, eg, first-cousin marriage. Clinical questionnaires (primary data form and follow-up data form) were developed to collect initial patient data and follow-up data at yearly intervals. RESULTS: Thirty-four centers provided clinical information for 254 patients and of these, 209 had sufficient data for analyses. When stratified by date of birth, 166 (79.4%) were born on or after January 1, 1990 (younger cohort) and 43 children (20.6%) were born before 1990 (older cohort). The gender distribution was equal in both cohorts. The median age at diagnosis was significantly later in the older cohort and no deaths were reported among these patients, suggesting that this group is biased toward long term survivors. In the younger cohort, 74.7% of the patients are alive, with a median age of 5.4 years. In this group, 40.5% of patients required ventilation and 11.6% developed chronic lung disease. Hypertension was a common, but not universal finding in both cohorts. The relative risk for developing hypertension was higher in the older cohort, but the median age at diagnosis was significantly earlier in the younger cohort. Chronic renal insufficiency (CRI) was reported in approximately 40% of patients with no significant difference in the relative risk between age groups. However, in the younger cohort, the median age at diagnosis was significantly earlier and the age of diagnosis of CRI and hypertension were significantly correlated. Clinically significant morbidities related to periportal fibrosis were more common in the older cohort. There was a trend toward increasing frequency of portal hypertension with age in both cohorts. Portal hypertension was not significantly correlated with either systemic hypertension or CRI. CONCLUSIONS: The ARPKD Clinical Database represents the largest single cohort of ARPKD patients collected to date. Our initial data analysis provides several new clinical insights. First, in our subset of long term survivors, ARPKD has a slower rate of disease progression, as assessed by age of ARPKD diagnosis, as well as age of diagnosis of clinical morbidities. Second, neonatal ventilation was strongly predictive of mortality as well as an earlier age of diagnosis in those who developed hypertension or chronic renal insufficiency. However, for infants who survive the perinatal period, the long term prognosis for patient survival is much better than generally perceived. Third, although systemic hypertension and CRI were significantly correlated with respect to age of diagnosis, similar relationships with portal hypertension were not evident, suggesting that disease progression may have organ-specific patterns. Fourth, only a subset of patients may be at risk for developing clinically significant manifestations of periportal fibrosis. Based on these observations, the next challenges will be to determine how various factors, such as specific mutations in the ARPKD gene, PKHD1(polycystic kidney and hepatic disease 1), variations in modifying gene loci, modulation by as yet unspecified environmental factors, and/or gene-environment interactions contribute to the marked variability in survival and disease expression observed among ARPKD patients. PMID- 12728093 TI - 2002 Job Lewis Smith Award acceptance address. PMID- 12728092 TI - Birth weight and blood cholesterol level: a study in adolescents and systematic review. AB - OBJECTIVE: To examine the relationship between birth weight and blood total cholesterol (TC) and to compare its strength with that of the relationship between current body mass index and TC. METHODS: 1). Cross-sectional study of adolescents, with retrospective ascertainment of birth weight from birth records or parental recall; 2). systematic review of studies examining the relations between birth weight and cholesterol at all ages. PARTICIPANTS: 1). 1532 individuals (92% white, 55% male) in 10 British towns; 2). 28 studies with 32 observations showing the change in TC per 1 kg increase in birth weight-6 in infancy, 14 in adolescents, 12 in adults. RESULTS: In the cross-sectional study, there was a weak inverse relation between birth weight and TC level (-.061 mmol/L fall in TC per kg increase in birth weight, 95% confidence interval -.131 to.008 mmol/L per kg) which was little affected by adjustment for current body size. The difference in TC corresponding to an interquartile range increase in birth weight (-.03 mmol/L) was approximately a quarter of that for an equivalent increase in body mass index (.11 mmol/L). In the systematic review, an inverse association between birth weight and TC of a similar size to that in the cross-sectional study was observed (-.048 mmol/L per kg, 95% confidence interval -.078 to -.018 mmol/L per kg) similar in strength at all ages. CONCLUSION: The relation of fetal nutrition to TC appears to be weak and is probably of limited public health importance when compared with the effects of childhood obesity. PMID- 12728094 TI - An innovative proposal for the health care financing system of the United States. PMID- 12728095 TI - Health care research on migrant farm worker children: why has it not had a higher priority? PMID- 12728096 TI - Fungal otitis externa--its association with fluoroquinolone eardrops. PMID- 12728098 TI - The 36-hour day: a wary commentary on the new accreditation council for graduate medical education guidelines on resident duty hours. PMID- 12728099 TI - Don't believe everything you read in the patient's chart. PMID- 12728100 TI - Unbound bilirubin in a term newborn with kernicterus. PMID- 12728101 TI - Oral health risk assessment timing and establishment of the dental home. AB - Early childhood dental caries has been reported by the Centers for Disease Control and Prevention to be perhaps the most prevalent infectious disease of our nation's children. Early childhood dental caries occurs in all racial and socioeconomic groups; however, it tends to be more prevalent in low-income children, in whom it occurs in epidemic proportions. Dental caries results from an overgrowth of specific organisms that are a part of normally occurring human flora. Human dental flora is site specific, and an infant is not colonized until the eruption of the primary dentition at approximately 6 to 30 months of age. The most likely source of inoculation of an infant's dental flora is the mother or another intimate care provider, through shared utensils, etc. Decreasing the level of cariogenic organisms in the mother's dental flora at the time of colonization can significantly impact the child's predisposition to caries. To prevent caries in children, high-risk individuals must be identified at an early age (preferably high-risk mothers during prenatal care), and aggressive strategies should be adopted, including anticipatory guidance, behavior modifications (oral hygiene and feeding practices), and establishment of a dental home by 1 year of age for children deemed at risk. PMID- 12728102 TI - Reimbursement for foods for special dietary use. AB - Foods for special dietary use are recommended by physicians for chronic diseases or conditions of childhood, including inherited metabolic diseases. Although many states have created legislation requiring reimbursement for foods for special dietary use, legislation is now needed to mandate consistent coverage and reimbursement for foods for special dietary use and related support services with accepted medical benefit for children with designated medical conditions. PMID- 12728103 TI - Facilities and equipment for the care of pediatric patients in a community hospital. AB - Many children who require hospitalization are admitted to community hospitals that are more accessible for families and their primary care physicians but vary substantially in their pediatric resources. The intent of this clinical report is to provide basic guidelines for furnishing and equipping a pediatric area in a community hospital. PMID- 12728104 TI - Partial ornithine transcarbamylase deficiency. PMID- 12728105 TI - The incidence of Kawasaki disease in the United States did not increase between 1988 and 1997. PMID- 12728106 TI - Behavior change counseling in the emergency department to reduce injury risk: a randomized, controlled trial. PMID- 12728107 TI - Tattooing and body piercing. PMID- 12728108 TI - Effectiveness of an educational intervention in modifying parental attitudes about antibiotic usage in children. AB - OBJECTIVE: To determine the effectiveness of educational materials in improving the attitudes of parents of young children about the judicious use of antibiotics. METHODS: We conducted a randomized controlled trial by recruiting parents of children who were younger than 24 months and being seen for any reason in primary care pediatric offices. At the time of enrollment, study parents indicated their level of agreement with 16 statements, including 9 statements about antibiotic usage and 7 about injury prevention. After being randomized, parents received either a pamphlet and a videotape promoting the judicious use of antibiotics (intervention group) or brochures about effective injury prevention (control group). Six weeks after enrollment, each group received another copy of the pamphlet or brochures and a follow-up questionnaire with the identical 16 statements. Responses on both questionnaires were transformed to an ordinal scale for analysis. Scores on the follow-up questionnaire for each statement about antibiotic use and injury prevention in the 2 groups were compared using linear regression, after controlling for the score obtained for the statement at enrollment. RESULTS: We enrolled a total of 499 eligible parents in the study; 358 (72%) completed the follow-up questionnaires. At study entry, there were no significant differences between parents in the intervention and control groups regarding attitudes for 15 of the 16 statements assessed. However, 6 weeks after receiving the antibiotic educational materials, parents in the intervention group had significantly different attitude scores for 5 of the 9 statements about the antibiotic use. In each case, the scores reflected attitudes that would promote the judicious use of antibiotics. We found significant attitudinal change for statements about the use of antibiotics for specific conditions in children; there were no differences between the 2 groups for more general or theoretical statements about antibiotic use. CONCLUSIONS: A simple educational effort was successful in modifying parental attitudes about the judicious use of antibiotics. Information about specific childhood conditions may be more effective in changing attitudes than more general information about antibiotic usage. PMID- 12728110 TI - Smoking patterns among adolescents with asthma attending upper secondary schools: a community-based study. AB - OBJECTIVE: Smoking among people who have asthma may be a serious health problem. We studied the prevalence of smoking and the relations between smoking and asthma, symptoms, medicine, and gender differences among adolescents with asthma. METHODS: A national cross-sectional study on health and lifestyles was performed in 1996-1997 using a computerized questionnaire in upper secondary schools in Denmark. We included 1887 pupils with asthma (defined as self-reported asthma diagnosed by a physician) and 20 688 controls. Smoking was categorized as daily, occasional, ex-smokers, and never smoked. We adjusted for age, gender, parents' job and smoking, family type, body mass index, and exercise habits. RESULTS: In total, 37.7% smoked currently and 16.5% smoked daily; more girls than boys smoked. More pupils with asthma than without smoked daily (adjusted odds ratio [OR]: 1.15; 95% confidence interval [CI]: 1.04-1.33), they smoked significantly more cigarettes daily (10.3 vs 9.6), and more were heavy smokers (> or =15 cigarettes daily; adjusted OR: 1.47; 95% CI:1.14-1.91). Furthermore, nearly twice as many pupils who had asthma with symptoms but were not using medicine smoked as pupils who had asthma without symptoms and were using medicine (adjusted OR: 1.84; 95% CI: 0.99-3.41). More boys with asthma than without started smoking before 14 years of age (adjusted OR: 1.75; 95% CI: 1.09-2.81). However, more pupils with asthma had tried to quit (adjusted OR: 1.26; 95% CI: 0.99-1.60). Pupils with and without asthma were occasional smokers and ex-smokers at a similar prevalence. CONCLUSIONS: More pupils with asthma than without smoked daily, and they also smoked more cigarettes per day. This is a major health concern, as adolescents have a high smoking prevalence in Denmark. PMID- 12728109 TI - Phototherapy use in jaundiced newborns in a large managed care organization: do clinicians adhere to the guideline? AB - OBJECTIVE: In 1994, the American Academy of Pediatrics (AAP) published a practice guideline with age-specific thresholds for phototherapy for healthy term newborns with hyperbilirubinemia. The purpose of this study was to examine adherence to the guideline in a large managed care organization. METHODS: We conducted a retrospective cross-sectional analysis of linked computerized databases from 11 Northern California Kaiser Permanente Medical Care Program hospitals. Newborn infants included were at least 37 weeks of gestation, had birth weights of at least 2500 g, and were born between January 1, 1995, and December 31, 1996. The primary outcome variable for the study was receipt of phototherapy according to the guideline. Total serum bilirubin (TSB) and infant age in hours at the time of bilirubin measurement were used to classify infants into 3 groups according to the AAP guideline: recommend phototherapy (R), consider phototherapy (C), and did not recommend phototherapy (N). Group R included infants with TSB levels of at least 15 mg/dL before 48 hours of age, at least 18 mg/dL before 72 hours, or at least 20 mg/dL after 72 hours. Group C included infants not in group R, with TSB levels of at least 12 mg/dL before 48 hours, 15 mg/dL before 72 hours, or at least 17 mg/dL after 72 hours. Group N included infants who were in neither group R nor group C and also did not have significant jaundice before 24 hours of age. Phototherapy codes from electronic databases were validated by chart review for a subset of 550 infants. RESULTS: Compared with chart review, phototherapy codes in the database were 94.4% sensitive (95% confidence interval [CI]: 89.1%-97.5%) and 100% specific (95% CI: 99.25%-100%). Among the 47 801 infants eligible, 2.3% received phototherapy. Phototherapy was administered to 54% of 1194 infants in group R (range across hospitals: 27%-77%), 16% of 2245 infants in group C (range: 5%-37%), and 0.2% of 44 362 infants in group N (range: 0.1%-0.6%). The predictors of phototherapy for group R, the group for whom phototherapy was recommended, determined by logistic regression were increasing TSB levels (odds ratio [OR]: 1.6/mg/dL; 95% CI: 1.4-1.7), reaching the AAP threshold at 24 to 47.9 hours of age compared with 48 hours or more (OR: 7.1; 95% CI: 4.3-11.9), gestational age of 37 weeks compared with 38 weeks or more (OR: 1.6; 95% CI: 1.1-2.3), age when phototherapy was first recommended (OR: 0.7/d; 95% CI: 0.6-0.8), and facility of birth (OR: 0.2-2.7). The facility of birth was a strong predictor of phototherapy use in all groups (R, C, and N). CONCLUSIONS: Clinicians provided phototherapy to only 54% of term infants with hyperbilirubinemia for whom it was recommended by the AAP. There is marked interhospital variation in phototherapy use in this large managed care system. Improved adherence to the guideline would require only a slight increase in the total rate of phototherapy use if unnecessary use for infants with lower levels of TSB were simultaneously decreased. PMID- 12728111 TI - The truth about language barriers: one residency program's experience. AB - OBJECTIVE: To describe perceptions of how a lack of house staff Spanish proficiency adversely affects communication with Spanish-speaking families with limited English proficiency (LEP). METHODS: An anonymous, structured questionnaire was administered to the house staff an of urban, university affiliated children's hospital that serves a population in which 10%-20% have LEP. RESULTS: Ninety-four percent (59 of 63) completed the questionnaire. Sixty eight percent (40 of 59) reported that they spoke little or no Spanish (although 36 of 40 expressed a desire to learn Spanish). Fifty-three percent (21 of 40) of these nonproficient residents reported that they used their inadequate language skills in the care of patients "often" or "every day." Many of these residents believed that LEP families under their care "never" or only "sometimes" understood their child's diagnosis (21 of 40), medications (11 of 40), discharge instructions (17 of 40), or follow-up plan (16 of 40). Eighty percent (32 of 40) admitted to avoiding communication with such families. Although all (40 of 40) agreed that hospital interpreters were effective, 30 of 40 nonproficient residents reported use of hospital interpreters "never" or only "sometimes." Fifty-three percent (21 of 40) of these nonproficient residents reported calling on their proficient colleagues "often" or "every day" for assistance. Thirty-two percent (19 of 59) of residents described themselves as "fluent" or "proficient" in Spanish. Fifty-eight percent (11 of 19) reported that they were asked to interpret for fellow residents "often" or "every day." Proficient residents estimated that they spent a mean of 2.3 hours per week interpreting for other residents. CONCLUSIONS: Despite a perception that they are providing suboptimal communication, nonproficient residents rarely use professional interpreters. Instead, they tend to rely on their own inadequate language skills, impose on their proficient colleagues, or avoid communication with Spanish-speaking families with LEP. PMID- 12728112 TI - Naturopathic treatment for ear pain in children. AB - OBJECTIVE: Otitis media is 1 of the most frequent diseases of early infancy and childhood and 1 of the most common reasons for children to visit a physician. In the past 2 decades, there has been a substantial increase in the diagnosis of otitis media worldwide. In the United States, 93% of all children have had at least 1 episode of acute otitis media (AOM) by 7 years of age. Otalgia is the hallmark of AOM. Most affected children either complain of earache or manifest behavior that the parents interpret as indicating ear pain. Treatment of the ear pain early in the course of AOM decreases both parental anxiety and the child's discomfort and accelerates the healing process. The objective of this study was to determine the efficacy and tolerability of naturopathic versus traditional treatment for the management of otalgia commonly associated with AOM in children. METHODS: The study was designed as a double-blind trial in an outpatient community clinic. A total of 171 children who were aged 5 to 18 years and had otalgia and clinical findings associated with middle-ear infection were studied. The children were randomly assigned to receive treatment with Naturopathic Herbal Extract Ear Drops (NHED) or anesthetic ear drops, with or without amoxicillin. On enrollment, the children were assigned by computer-numbered randomization to receive NHED (contents: allium sativum, verbascum thapsus, calendula flores, hypericum perfoliatum, lavender, and vitamin E in olive oil) 5 drops 3 times daily, alone (group A) or together with a topical anesthetic (amethocaine and phenazone in glycerin) 5 drops 3 times daily (group B), or oral amoxicillin 80 mg/kg/d (maximum 500 mg/dose) divided into 3 doses with either NHED 5 drops 3 times daily (group C) or topical anesthetic 5 drops 3 times daily (group D). A double-blind design was used, and all ear drops were placed in identical bottles. Treatment was initiated by the nurse in all cases. A single physician (M.S.) evaluated and treated all of the patients included in the study and recorded all of the data. The presence or absence of ear pain was assessed over 3 days with a visual analog scale. Ear pain was assessed by a specially devised observational instrument based on previous reports. One side of the instrument consisted of a linear numbered scale, from 1 (no pain) to 10 (worst possible pain), and a corresponding color scale, ranging from blue to dark red. The reverse side contained a scale of 5 facial expressions, ranging from broad smile (no pain) to a sad and crying face (worst possible pain), and a corresponding color scale, ranging from blue to dark red. RESULTS: There were no significant between-group differences in patient age or gender, degree of fever, main symptoms, associated symptoms, and severity or laterality of acute otitis media. Each group had a statistically significant improvement in ear pain over the course of the 3 days. Patients who were given ear drops alone had a better response than patients who were given ear drops together with amoxicillin. Results were better in the NHED group than in the controls. Nevertheless, the findings indicated that the pain was mostly (80%) self-limited and could be explained simply by the time elapsed. The American Academy of Otolaryngology-Head and Neck Surgery guidelines recommend topical medications as the first line of treatment for ear pain in the absence of systemic infection or serious underlying disease. Because no evidence was found that systemic antibiotics alone improved treatment outcome, if antibiotics do not change the natural course of otitis media, then the main goal of treatment, as in the present study, should be to alleviate the ear pain. The alternative, naturopathic herbal extract medications, may offer many new possibilities in the management of ear pain associated with AOM. Primary care physicians should be aware that at least 10% of their patients may have tried 1 or more forms of alternative/complementary medicine before presenting for consultation. As it was widely reported in the medical literature, these herb, these herbal extracts have the potential to meet all of the requirements of appropriate medication that could be routinely used in the pediatric patient, namely in vitro bacteriostatic and bacteriocidal activity against common pathogens, immunostimulation ability, antioxidant activity, and anti-inflammatory effects. They are also well-absorbed with good penetration into the tissue surrounding the tympanic membrane. They have been found to enhance local immunologic activity. Finally, herbal extracts are well-tolerated (owing to their long elimination time), easy to administer, and less expensive than the new antibiotics. There are no documented side effects. On the basis of our findings that the group with the most significant treatment effects (NHED with topical anesthetic) explained only 7.3% of the total pain reduction, we propose that sometimes the general practitioner or pediatrician needs to give the human body a chance to repair itself. Nevertheless, if the physician believes that there is an indication for some treatment, especially if the parents are anxious, then a local treatment such as one used in our study might be adequate. CONCLUSIONS: This study suggests that in cases of ear pain caused by AOM in children in which active treatment, besides a simple 2- to 3-day waiting period, is needed, an herbal extract solution may be beneficial. Concomitant antibiotic treatment is apparently not contributory. PMID- 12728114 TI - Paranasal sinus findings in children during respiratory infection evaluated with magnetic resonance imaging. AB - OBJECTIVE: The spreading of acute respiratory infection into the paranasal sinuses in children is poorly defined. The main objective of this study was to evaluate the frequency and spontaneous resolution of paranasal sinus abnormalities in children with acute respiratory infection using magnetic resonance imaging (MRI). METHODS: We examined 60 children with MRI (mean age: 5.7 years) with symptoms of acute respiratory infection. Twenty-six children with major abnormalities in the first MRI scan had a follow-up MRI taken 2 weeks later. RESULTS: The children had had symptoms of uncomplicated acute respiratory infection for an average of 6 days before the first examination (mean duration: 6.5; standard deviation: 3.0). Approximately 60% of the children had major abnormalities in their maxillary and ethmoidal sinuses, 35% in the sphenoidal sinuses, and 18% in the frontal sinuses. The most common abnormal finding was mucosal swelling. The mean overall MRI scores correlated significantly with the symptom scores (r(s) = 0.3). Of the individual symptoms, nasal obstruction, nasal discharge, and fever were significantly related to the MRI scores. Among the 26 children with major abnormalities in the first MRI, the findings subsequently improved significantly (mean [standard deviation] score: 12.7 [5.6] to 5.7 [5.2]), irrespective of the resolution of symptoms. CONCLUSIONS: These observations indicate that acute respiratory infection mostly spreads into the paranasal sinuses of children in the form of mucosal edema and that these abnormalities tend to resolve spontaneously without antimicrobial treatment. PMID- 12728113 TI - Breastfeeding, exposure to organochlorine compounds, and neurodevelopment in infants. AB - OBJECTIVE: Exposure to organochlorine compounds (OCs) occurs both in utero and through breastfeeding. Levels of hexachlorobenzene (HCB) found in the cord serum of newborns from a population located in the vicinity of an electrochemical factory in Spain were among the highest ever reported. We studied the association between exposure to OCs and breastfeeding on neurodevelopment in the 1-year-old infants of this population. METHODS: A birth cohort including 92 mother-infant pairs was recruited between 1997 and 1999 in 5 neighboring villages (84% of possible recruits). The mental and psychomotor development of each infant was assessed at 13 months using the Bayley and the Griffiths Scales of Infant Development. OCs were measured in cord serum. RESULTS: Dichlorodiphenyl dichloroethylene (p,p'DDE) cord serum levels were negatively associated with both mental and psychomotor development. For each doubling of a dose of p,p'DDE, we found a resultant decrease of 3.50 points (standard error: 1.39) on the mental scale and 4.01 points (standard error: 1.37) on the psychomotor scale. Exposure to polychlorinated biphenyls was only marginally associated with psychomotor development. Prenatal exposure to HCB had no effect on child neurodevelopment. Long-term breastfeeding was associated with better performance on both the mental and motor scales. Short-term breastfed infants with higher p,p'DDE levels in cord serum were associated with the lowest scores on both the mental and the psychomotor scales. CONCLUSIONS: Prenatal exposure to p,p'DDE was associated with a delay in mental and psychomotor development at 13 months. No association was found for exposure to HCB. Long-term breastfeeding was found to be beneficial to neurodevelopment, potentially counterbalancing the impact of exposure to these chemicals through breast milk. PMID- 12728115 TI - Risk factors for intraventricular hemorrhage in very low birth weight premature infants: a retrospective case-control study. AB - OBJECTIVE: High-grade intraventricular hemorrhage (IVH) is an important cause of severe cognitive and motor neurologic impairment in very low birth weight infants and is associated with a high mortality rate. The risk of IVH is inversely related to gestational age and birth weight. Previous studies have proposed a number of risk factors for IVH; however, lack of adequate matching for gestational age and birth weight may have confounded the results. The purpose of this study was to identify variables that affect the risk of high-grade IVH, using a retrospective and case-control clinical study. METHODS: From a cohort of 641 consecutive preterm infants with a birth weight of <1500 g, 36 infants with IVH grade 3 and/or 4 were identified. A control group of 69 infants, closely matched for gestational age and birth weight, was selected. Maternal factors, labor and delivery characteristics, and neonatal parameters were collected in both groups. Results of cranial ultrasound examinations, whether routine or performed in presence of clinical suspicion, were also collected. Univariate analysis and multivariate logistic regression analysis were performed. RESULTS: High fraction of inspired oxygen in the first 24 hours, pneumothorax, fertility treatment (mostly IVF), and early sepsis were associated with an increased risk of IVH. A higher number of suctioning procedures, a higher first hematocrit, and a relatively low arterial pressure of carbon dioxide during the first 24 hours of life were associated with a lower occurrence. In the multivariate logistic regression model, early sepsis (odds ratio [OR]: 8.19; 95% confidence interval [CI]: 1.55-43.1) and fertility treatment (OR: 4.34; 95% CI: 1.42-13.3) were associated with a greater risk of high-grade IVH, whereas for every dose of antenatal steroid treatment there was a lower risk of high-grade IVH (OR: 0.52; 95% CI: 0.30-0.90) and each decrease in a mmHg unit of arterial pressure of carbon dioxide during the first 24 hours was associated with a lower risk of IVH (OR: 0.91; 95% CI: 0.83-0.98). This multivariate model had a sensitivity of 77%, a specificity of 75%, and a positive predictive value of 76%. The area under the curve derived from the receiver operator characteristic plots is 0.82. CONCLUSIONS: Our results confirm that the development of IVH is associated with early sepsis and failure to give antenatal steroid treatment. We propose that fertility treatment (and especially IVF) may be a new risk factor, and more research is needed to assess its role. PMID- 12728117 TI - Rapid increase in grip force after start of pamidronate therapy in children and adolescents with severe osteogenesis imperfecta. AB - OBJECTIVE: To examine changes in grip force during pamidronate therapy in children and adolescents with severe osteogenesis imperfecta (OI). METHODS: Maximal isometric grip force of the nondominant hand was prospectively determined in 42 patients (age at the start of the study: 7.3-15.9 years; 18 girls) with severe forms of OI. Patients were treated with intravenous pamidronate infusions given in 4 monthly cycles, each cycle consisting of 3 infusions (1 mg pamidronate/kg body wt) on 3 successive days. RESULTS: At the start of pamidronate therapy, grip force was low compared with age-specific reference data (age z score mean +/- standard deviation: -2.7 +/- 2.1) but was normal for weight (weight z score: -0.1 +/- 1.8). Four months after the first pamidronate infusion cycle, grip force had increased significantly, whether related to age (age z score: -2.0 +/- 1.8) or to weight (weight z score: 0.6 +/- 1.5). At 2 years after the start of therapy, grip force z scores were not significantly different from the 4-month results. CONCLUSIONS: Maximal isometric grip force markedly increases after a single cycle of intravenous pamidronate in children with severe forms of OI, and this gain in grip force is maintained for at least 2 years. PMID- 12728116 TI - Underestimation of infant mortality rates in one republic of the former Soviet Union. AB - OBJECTIVES: Kazakhstan's live-birth definition--that dates from the former Soviet Union (FSU) era--differs from that used by the World Health Organization (WHO). We studied the impacts of both live-birth definitions on the computations of the infant mortality rate (IMR) and maternal and child health (MCH) planning in Zhambyl Oblast, Kazakhstan. METHODS: We interviewed caregivers and abstracted medical records to obtain birth weight and age-at-death information on infant deaths in Zhambyl Oblast from November 1, 1996, through October 31, 1997. Using the 2 indicators of birth weight and age at death, we created a matrix delineating the respective contribution to infant death (maternal health, newborn care, or infant care) for the cells. We then calculated the IMR, birth weight specific IMR (BWS-IMR), and birth weight-proportionate IMR (BWP-IMR) for each cell. RESULTS: The observed IMR in Zhambyl Oblast, in 1996--using the definition of a live birth from the FSU--was 32 per 1000 live births. The recalculated IMR- using the WHO definition--was 58.7 per 1000 live births. Computed estimates of the contribution to infant death, by the categories of maternal health, newborn care, and infant care, were 10%, 23%, and 67%, respectively, when using the live birth definition from the Soviet era. These estimates shifted to 24%, 41%, and 35%, respectively, when using the WHO definition, yet only 8% of the Zhambyl Oblast MCH budget was earmarked to maternal health and newborn care, which we estimated accounted for 65% of infant deaths. CONCLUSIONS: The live-birth definition commonly used in the FSU underestimated the IMR and undervalued the contributions to infant death by both maternal health and newborn care. We recommend that all republics of the FSU adopt the WHO live-birth definition so that the IMR can serve as a better indicator for MCH planning. PMID- 12728118 TI - Trends in referral to a single encopresis clinic over 20 years. AB - OBJECTIVE: To compare the characteristics of children with encopresis referred to a single encopresis clinic over the course of 20 years, including symptoms, previous diagnostic and therapeutic interventions, and parental attitudes. METHODS: A retrospective study was conducted of an encopresis clinic at a tertiary care pediatric hospital. Questionnaires at initial evaluation elicited information about bowel habits, soiling, previous evaluations, previous treatments, and parental attitudes. RESULTS: In 503 children with encopresis, the average age of referral dropped from 115 months during the earliest 5 years to 77 months during the most recent 5 years. Children who had soiling for >3 years before referral decreased from 63% to 12%. The use of barium enema before referral decreased from 14% to 5%, as did psychological evaluation, from 25% to 14%. Previous therapy with enemas decreased from 45% to 27%. Mineral oil use remained at approximately 50%, and 20% of children had no previous treatment. Symptoms at referral and parental attitudes did not change across the years. CONCLUSIONS: Children are now referred at an earlier age to our tertiary encopresis clinic. The number of invasive and psychological evaluations has decreased before referral. However, treatment by many primary care providers before the referral has not changed. These data may suggest that pediatricians have increased awareness of encopresis and greater appreciation of its primarily physical rather than psychological nature. Additional studies will be needed to determine how these factors affect outcome. PMID- 12728119 TI - Risk factors for tuberculosis infection in children in contact with infectious tuberculosis cases in the Gambia, West Africa. AB - OBJECTIVE: Tuberculosis (TB) infection is highly prevalent in developing countries. As infected children represent a large proportion of the pool from which TB cases will arise, knowledge of the factors that influence TB infection in children are of importance to evaluate transmission of infection in the community and adapt TB control activities. There are limited data on the risk of infection in child populations in developing countries. METHODS: We performed a household contact study in The Gambia (West Africa), in which children who were living in contact with individuals who had proven smear-positive pulmonary TB cases were investigated. A questionnaire was addressed to the mother or caregiver of the child to investigate the presence of various risk factors and assess the degree of exposure of the child to the individual with TB within the household. A tuberculin skin test (TST) was performed on each child. TST sizes > or =5 and 10 mm, respectively, were considered positive. RESULTS: Households of 206 TB cases were visited, and 384 children aged <5 years were examined. The median age was 2, and 48% were girls. The distribution of TST responses followed a bimodal pattern, with 135 (35%) children presenting a palpable induration. Random effects logistic regression analysis demonstrated that the risk of positive TST response in the child increased with the geographic proximity of the child to the individual with TB within the household and with the degree of activities shared with the individual with TB. It was also associated with the clinical severity of the disease in the index case. Nutritional status and presence of a bacille Calmette Guerin (BCG) scar were not independent risk factors for TST positivity in this population. On multivariate analysis, the effect of geographic proximity to the individual with TB, household size, and duration of cough in the index case persisted for TST responses > or =5 mm. CONCLUSIONS: In a highly endemic country with high BCG vaccination coverage in Africa, TB infection in children who were in contact with individual with infectious TB was directly related to the intensity of exposure of the child to the individual with TB. Our data suggest that a positive TST in a child reflects most probably TB infection rather than previous BCG vaccination. Contact tracing can play a major role in the control of TB in developing countries. PMID- 12728120 TI - Race/ethnicity and asthma among children presenting to the emergency department: differences in disease severity and management. AB - OBJECTIVE: To investigate racial/ethnic differences in acute asthma among children who present to the emergency department (ED). METHOD: We analyzed data from 2 prospective cohort studies performed during 1997-1998 as part of the Multicenter Airway Research Collaboration. Using a standardized protocol, researchers at 40 EDs in 18 US states provided 24-hour-per-day coverage for a median of 2 weeks per year. Children with acute asthma were interviewed in the ED and by telephone 2 weeks after discharge. RESULTS: Among 1095 patients, 679 (62%) were black, 256 (23%) were Hispanic, and 160 (15%) were white. Black and Hispanic children had greater histories of lifetime (63%, 64%, 46%) and past-year (34%, 31%, 14%) hospitalization and more ED visits in the past year (medians: 2, 3, 1). Asthma severity at ED presentation, ED management and course, hospitalization during the index visit, discharge prescriptions, and postdischarge outcomes were equivalent among all race/ethnic groups. CONCLUSION: Despite pronounced race/ethnicity-based differences in chronic asthma, all racial/ethnic groups exhibited similar acute asthma severity, ED management, and course. However, given that black and Hispanic children exhibited much higher admission histories and past ED use, the equivalence in inhaled corticosteroid prescriptions on discharge is a disconcerting pattern that mirrors previous literature on outpatient prescription practices. In addition to barriers attributable to socioeconomic factors, health care providers and policy makers should target equalizing deficiencies in preventive medication prescription practices. PMID- 12728121 TI - Autoimmunity in Wiskott-Aldrich syndrome: risk factors, clinical features, and outcome in a single-center cohort of 55 patients. AB - OBJECTIVES: To evaluate the occurrence of autoimmune and inflammatory complications in Wiskott-Aldrich syndrome (WAS) and to determine risk factors and the prognosis of such complications with the aim of improving the definition of treatment options. METHODS: We reviewed the records of 55 patients with WAS evaluated at Necker-Enfants Malades Hospital (Paris) from 1980 to 2000. RESULTS: Forty patients (72%) had at least 1 autoimmune or inflammatory complication. Autoimmune hemolytic anemia was detected in 20 cases (36%); in all cases, onset occurred before the age of 5 years. Other complications included neutropenia (25%), arthritis (29%), skin vasculitis (22%), cerebral vasculitis (7%), inflammatory bowel disease (9%), and renal disease (3%). The median survival of the entire population was 14.5 years. Two autoimmune complications and 1 biological factor were predictive of a poor prognosis in this population: autoimmune hemolytic anemia, severe thrombocytopenia recurring after splenectomy, and high serum immunoglobulin M (IgM) levels before splenectomy. Autoimmune hemolytic anemia was significantly more observed in patients with high serum IgM level. CONCLUSIONS: High serum IgM concentration before splenectomy was identified as a risk factor for autoimmune hemolytic anemia; however, it must be confirmed. Autoimmune hemolytic anemia and severe thrombocytopenia recurring after splenectomy were 2 indicators of a poor prognosis. Those results suggest that patients with WAS and IgM levels more than mean + 2 standard deviations before splenectomy should be placed under strict surveillance. Furthermore, severe autoimmune complications should lead, as early as possible, to hematopoietic stem cell transplantation using the best available donor. PMID- 12728122 TI - Medication use in the treatment of pediatric insomnia: results of a survey of community-based pediatricians. AB - OBJECTIVES: To examine clinical practice patterns, beliefs, and attitudes regarding the use of both nonprescription and prescription medications by community-based pediatricians for children with significant difficulties in initiating and/or maintaining sleep. METHODS: A survey was mailed to 3424 American Academy of Pediatrics members in 6 US cities. RESULTS: The final sample (n = 671) consisted of practitioners who identified themselves as primary care pediatricians. Three percent +/- 7% of visits in the respondents' practices were for pediatric insomnia, although there was a wide range in the numbers of children identified during a typical 6-month practice period. More than 75% of practitioners had recommended nonprescription medications, and >50% had prescribed a sleep medication. Specific clinical circumstances in which medications were most commonly used were acute pain and travel, followed by children with special needs (mental retardation, autism, and attention deficit/hyperactivity disorder). Antihistamines were the most commonly reported nonprescription medications for sleep. Melatonin or herbal remedies had been recommended by approximately 15% of the respondents. alpha-agonists were the most frequently prescribed sleep medications (31%). The likelihood of prescribing medication for sleep was 2- to 4-fold greater in respondents who treated children with attention-deficit/hyperactivity disorder for daytime behavioral problems or nocturnal sleep problems, respectively. Practitioners expressed a range of concerns about sleep medication appropriateness, safety, tolerance, and side effects in children. CONCLUSIONS: The practice of prescribing or recommending sedatives and hypnotics for pediatric insomnia is common among community-based pediatricians, especially among special needs patients. An empirically based approach to the use of these medications is needed. PMID- 12728123 TI - Hemophagocytic lymphohistiocytosis masquerading as child abuse: presentation of three cases and review of central nervous system findings in hemophagocytic lymphohistiocytosis. AB - Hemophagocytic lymphohistiocytosis (HLH) is a rare disease resulting from abnormal proliferation of histiocytes in tissues and organs. Although the disease generally presents with systemic symptoms such as pancytopenia, coagulopathy, and organomegaly, HLH may also present with central nervous system (CNS) manifestations. CNS events can range from irritability to encephalopathy and coma. Retinal and intracranial hemorrhages are among the neuropathologic findings in these children. Patients who present with CNS findings may have symptoms that mimic those of inflicted injury. These children are at risk, therefore, for misdiagnosis as victims of child abuse. Such an error causes not only unnecessary additional trauma to the family but also, more important, a delay in initiating effective therapy. We present 3 cases of children with HLH who initially came to medical attention with neurologic findings, all suspected to be victims of child abuse. Subsequent laboratory evaluations, however, were consistent with the diagnosis of HLH. No additional evidence of child abuse was obtained, and the charges eventually were dropped. Two of the 3 children died from their disease shortly after presentation; the third is surviving with no evidence of HLH several months after allogeneic bone marrow transplantation. Although the diagnosis of child abuse certainly is all too common, clinicians need to be diligent and informed to avoid assigning this label erroneously. Several laboratory findings of HLH may alert physicians to the possibility of this diagnosis. The timely diagnosis of and institution of therapy for HLH may reduce ultimate morbidity and mortality. PMID- 12728124 TI - An integrative perspective: mothers' influence on child health preconceptionally, prenatally, and in early childhood. PMID- 12728125 TI - Reducing the risk of alcohol-exposed pregnancies: a study of a motivational intervention in community settings. AB - OBJECTIVES: To test the feasibility and impact of a motivational intervention in reducing drinking and/or increasing effective contraception in women who are at risk for an alcohol-exposed pregnancy. METHODS: A multisite single-arm pilot study was conducted in 6 community settings in 3 large cities. A total of 2384 women were screened for eligibility; 230 were eligible on the basis of their alcohol use and lack of contraception. Of the eligible women, 190 consented and were enrolled, and 143 (75.3%) completed the 6-month follow-up. The intervention consisted of 4 manual-guided motivational counseling sessions delivered by mental health clinicians and 1 contraceptive counseling session delivered by a family planning clinician. Outcome measures include intervention completion rates, alcohol use (frequency, quantity, and bingeing), contraceptive use and effectiveness, and risk for alcohol-exposed pregnancy. RESULTS: Among women who completed the 6-month follow-up, 68.5% were no longer at risk of having an alcohol-exposed pregnancy; 12.6% of women who completed the program reduced drinking only; 23.1% used effective contraception only; and 32.9% reported both. Results were consistent across the 6 diverse high-risk settings. CONCLUSIONS: This study provides evidence that providing 4 sessions of motivational interviewing plus a contraception counseling session is feasible and strongly suggests that this intervention can decrease the risk of alcohol-exposed pregnancy in women in high-risk settings. Additional investigation in a randomized controlled trial is warranted. PMID- 12728126 TI - Binge drinking in the preconception period and the risk of unintended pregnancy: implications for women and their children. AB - OBJECTIVE: To assess the relationship between unintended pregnancy resulting in a live birth and binge drinking (having 5 or more alcoholic beverages on 1 occasion) in the 3 months before pregnancy (the preconception period) and to characterize women who are of childbearing age and binge drink. METHODS: A case control study was conducted of women with pregnancies that resulted in a live birth, comparing those with unintended pregnancies with those with intended pregnancies. Data analyzed were from the 15 states that participated in the Pregnancy Risk Assessment Monitoring System from 1996-1999. RESULTS: Of 72 907 respondents, 45% of pregnancies were unintended. Compared with women with intended pregnancy, women with unintended pregnancy were more likely to be young and black and to report preconception binge drinking (16.3% vs 11.9%; odds ratio [OR]: 1.43; 95% confidence interval [CI]: 1.13-1.54). After adjusting for potential confounders, preconception binge drinking was associated with unintended pregnancy for white women (adjusted OR: 1.63; 95% CI: 1.47-1.80) but not for black women (adjusted OR: 0.96, 95% CI: 0.77-1.20). Overall, 14% of women reported preconception binge drinking. Women who binge drank in the preconception period were more likely to be white and unmarried; to smoke and be exposed to violence in the preconception period; and to consume alcohol, binge drink, and smoke during pregnancy. CONCLUSIONS: Binge drinking in the preconception period was associated with unintended pregnancies resulting in a live birth among white women but not among black women. Preconception binge drinkers were more likely to engage in other risky behaviors, including drinking during pregnancy. Comprehensive interventions to reduce binge drinking may reduce unintended pregnancies, as well as other adverse maternal and pediatric health outcomes. PMID- 12728127 TI - Pregnancy intendedness and the use of periconceptional folic acid. AB - OBJECTIVE: Periconceptional use of folic acid can prevent birth defects, including at least 50% of neural tube defects. This study used an ongoing surveillance system to explore the association between pregnancy intendedness and women taking periconceptional folic acid. METHODS: Oregon Pregnancy Risk Assessment Monitoring System (PRAMS) surveys a stratified random sample of women after a live birth. In 1998-1999, 1867 women completed the survey (64.0% response rate); responses were weighted for nonresponse. Women were asked whether they took folic acid most days in the month before becoming pregnant. RESULTS: Overall, 33.2% of women took folic acid most days in the month before becoming pregnant, and 39.9% said that their pregnancy was unintended. Adolescent mothers were less likely to take periconceptional folic acid (9.2%) and more likely to report unintended pregnancy (62.0%) than older women. Overall, women who said that their pregnancy was intended were more likely to report that they had taken periconceptional folic acid (odds ratio: 4.75; 95% confidence interval: 3.16 7.14); after controlling for maternal age and income the odds ratio was 3.70 (95% confidence interval: 2.38-5.56). CONCLUSIONS: Women whose pregnancies were intended were more likely to have been taking periconceptional folic acid than women whose pregnancies were unintended. The importance of fertile women's taking daily multivitamins that contain 400 microg (0.4 mg) of folic acid should be stressed among women who are not contemplating pregnancy, especially adolescents and low-income women. PMID- 12728128 TI - Do multivitamin supplements attenuate the risk for diabetes-associated birth defects? AB - OBJECTIVE: To evaluate whether the risk for birth defects associated with maternal diabetes is attenuated by use of multivitamin supplements during the periconceptional period. METHODS: In the population-based Atlanta Birth Defects Case-Control Study, we identified case infants who had nonsyndromic birth defects that were reported to be associated with diabetes (n = 3278) and were born during 1968-1980 to residents of metropolitan Atlanta. Controls were infants without birth defects (n = 3029). Maternal diabetes was defined as reported diabetes with onset before the date of birth of the index infant, and periconceptional use of multivitamins was defined as reported regular use of multivitamin supplements from 3 months before pregnancy through the first 3 months of pregnancy. RESULTS: Offspring of mothers with diabetes had an increased risk for selected birth defects. However, the increased risk was limited to offspring of mothers who had diabetes and had not taken multivitamins during the periconceptional period (odds ratio: 3.93; 95% confidence interval: 1.79-8.63). Offspring of mothers who had diabetes and had taken multivitamins during the periconceptional period had no increased risk for birth defects (odds ratio: 0.15; 95% confidence interval: 0.00 1.99). CONCLUSIONS: Periconceptional use of multivitamin supplements may reduce the risk for birth defects among offspring of mothers with diabetes. PMID- 12728129 TI - Maternal obesity and risk for birth defects. AB - OBJECTIVE: Several studies have shown an increased risk for neural tube defects associated with prepregnancy maternal obesity. Because few recent studies have examined the relation between maternal prepregnancy obesity and overweight and other birth defects, we explored the relation for several birth defects and compared our findings with those of previous studies. METHODS: We conducted a population-based case-control study of several selected major birth defects using data from the Atlanta Birth Defects Risk Factor Surveillance Study. Mothers who delivered an infant with and without selected birth defects in a 5-county metropolitan Atlanta area between January 1993 and August 1997 were interviewed. Maternal body mass index (BMI) was calculated from self-reported maternal prepregnancy weight and height. Women with known preexisting diabetes were excluded. The risks for obese women (BMI > or =30) and overweight women (BMI 25.0 29.9) were compared with those for average-weight women (BMI 18.5-24.9). RESULTS: Obese women were more likely than average-weight women to have an infant with spina bifida (unadjusted odds ratio [OR]: 3.5; 95% confidence interval [CI]: 1.2 10.3), omphalocele (OR: 3.3; 95% CI: 1.0-10.3), heart defects (OR: 2.0; 95% CI: 1.2-3.4), and multiple anomalies (OR: 2.0; 95% CI: 1.0-3.8). Overweight women were more likely than average-weight women to have infants with heart defects (OR: 2.0; 95% CI: 1.2-3.1) and multiple anomalies (OR: 1.9; 95% CI: 1.1-3.4). CONCLUSIONS: Our study confirmed the previously established association between spina bifida and prepregnancy maternal obesity and found an association for omphalocele, heart defects, and multiple anomalies among infants of obese women. We also found an association between heart defects and multiple anomalies and being overweight before pregnancy. A higher risk for some birth defects is yet another adverse pregnancy outcome associated with maternal obesity. Obesity prevention efforts are needed to increase the number of women who are of healthy weight before pregnancy. PMID- 12728130 TI - Trends in multiple births conceived using assisted reproductive technology, United States, 1997-2000. AB - OBJECTIVE: To examine trends in multiple births conceived using assisted reproductive technology (ART) in the United States between 1997 and 2000 and to estimate the proportion of all US multiple births attributable to ART use. METHODS: We analyzed population-based data of 109 519 live-born infants who were conceived in the United States using ART and born between 1997 and 2000 and population-based data of 15 856 809 live-born infants who were delivered in the United States between 1997 and 2000. Multiple birth rates (the number of live born infants delivered in multiple gestation pregnancies per 1000 live births) and the proportion of all US multiple births attributable to ART were evaluated. RESULTS: The twin rate for ART patients increased between 1997 and 2000, reaching 444.7 per 1000 live births in 2000, whereas the triplet/+ rate declined substantially from 134.3 to 98.7 per 1000 live births from 1997-2000. From 1997 2000, the proportion of multiple births in the United States attributable to ART increased from 11.2% to 13.6%, whereas the proportion attributable to natural conception decreased from 69.9% to 64.5%. In 2000, the proportion of triplet/+ births attributable to ART and to natural conception was 42.5% and 17.7%, respectively. The contribution of ART to multiple births increased substantially with maternal age, from 11.6% for triplet/+ infants born to women aged 20 to 24 to 92.8% for women aged 45 to 49 years. CONCLUSIONS: The contribution of ART to twin birth rates continues to increase, but the contribution of ART to triplet/+ birth rates has declined. PMID- 12728132 TI - Sharing prescription medication among teenage girls: potential danger to unplanned/undiagnosed pregnancies. AB - OBJECTIVE: The objective of this study was to determine how often children and adolescents share prescription medications and, because of teratogenic concerns, assess specific reasons why girls might engage in medication-sharing behaviors. METHODS: Data were collected as part of Youthstyles, a mail survey of children and adolescents 9 through 18 years of age (764 girls and 804 boys) about health issues, attitudinal variables, and media preferences. Information collected by the survey included the respondent's history of borrowing or sharing prescription medications, the frequency with which sharing occurred, the reasons why medications might be borrowed or shared, and who influences their decisions to borrow or share medication. RESULTS: A total of 20.1% of girls and 13.4% of boys reported ever borrowing or sharing medications. Of the girls surveyed, 15.7% reported borrowing prescription medications from others, and 14.5% reported sharing their prescription medication with someone else. The reported likelihood of sharing increased with age. Medication sharing or borrowing was not a "one time only" emergency use for many: 7.3% of girls 15 through 18 years of age had shared medications >3 times. Reasons that girls gave for why they would share medications included having a prescription for the same medicine (40.2%), getting the medication from a family member (33.4%), having the same problem as the person who had the medication (29%), or wanting something strong for pimples or oily skin (10.5%). CONCLUSIONS: Medication sharing is relatively common among children and adolescents and is more common among girls than boys. An adolescent who receives a medication via sharing does not receive the appropriate information about its actions and possible negative interactions with other medications or any other associated risks. Sharing potentially teratogenic drugs is of special concern. Many barriers exist to communicating the risk about teratogenic drugs to women and girls, particularly if they are not planning a pregnancy or are unaware that they are already pregnant. These findings suggest the need for basic research on issues related to the dangers of medication sharing and teratogenic risks, as well as the development of successful approaches to communicate these risks. PMID- 12728131 TI - Fertility treatments and craniosynostosis: California, Georgia, and Iowa, 1993 1997. AB - OBJECTIVE: Craniosynostosis, a malformation caused by premature closure of 1 or more cranial sutures, is a rare birth defect usually of unknown cause; however, it is often associated with advanced maternal age. Because fertility treatments are also associated with increased maternal age, this study investigated the possible association between fertility treatments and craniosynostosis. METHODS: Data from the Birth Defect Risk Factor Surveillance study were used, which included infants who were delivered from 1993 through 1997 in California, Georgia, and Iowa. Cases were defined as infants who had nonfamilial, nonsyndromic craniosynostosis and were ascertained through existing birth defect surveillance systems. Controls, infants without birth defects, were selected from the same regions and time period. RESULTS: Mothers of 99 case infants and 777 control infants from the 3 study locations participated in this study by completing a telephone interview. Unadjusted analyses showed associations with craniosynostosis for mothers who had used clomiphene citrate (odds ratio[OR]: 3.8; 95% confidence interval [CI]: 1.1-12.3), artificial insemination (OR: 4.2; 95% CI: 0.8-9.4), or assisted reproductive techniques (OR: 4.2; 95% CI: 0.5 27.3). CONCLUSIONS: This is the first study that has found associations between fertility treatments and craniosynostosis. However, the numbers are small; therefore, the results should be viewed with caution. PMID- 12728133 TI - Associations of intrauterine growth restriction among term infants and maternal pregnancy intendedness, initial happiness about being pregnant, and sense of control. AB - OBJECTIVE: Term infants (> or =37 weeks' gestation) who weigh <2500 g have intrauterine growth restriction (IUGR) and have a higher risk of mortality and morbidity. Little is known about how psychosocial factors affect the risk of IUGR. We examined the association between IUGR and maternal pregnancy intendedness, initial happiness about becoming pregnant, and maternal sense of control. METHODS: We analyzed data from a survey of California mothers aged > or =15 years with term live births in 1999 and 2000 (N = 5961). Mothers were asked about pregnancy intendedness before pregnancy, initial happiness about becoming pregnant, and maternal sense of control, assessed by a standardized scale. We examined the association of having an infant with IUGR and these factors in univariate and multivariate analyses. RESULTS: Mothers with low sense of control (3.0%) and average sense of control (2.7%) were more likely to have an infant with IUGR than mothers with high sense of control (1.8%; odds ratio: 1.8; 95% confidence interval: 1.2-2.9; and odds ratio: 1.6; 95% confidence interval: 1.0 2.5). After multivariate analysis, we found no significant association between sense of control and IUGR. We also found no significant association between IUGR and pregnancy intendedness and happiness about becoming pregnant. CONCLUSIONS: We found no statistically significant association between IUGR and maternal pregnancy intendedness, initial happiness about becoming pregnant, and maternal sense of control. Although research should continue to explore associations between psychosocial factors and IUGR, prenatal care programs should focus on known risk factors for IUGR. PMID- 12728134 TI - Trends and variations in smoking during pregnancy and low birth weight: evidence from the birth certificate, 1990-2000. AB - OBJECTIVE: This study compares patterns of tobacco use during pregnancy over time and across population subgroups and examines the impact of maternal smoking on the incidence of low birth weight (LBW). The study also evaluates the use of birth certificates to monitor prenatal smoking. METHODS: The birth certificates of all states (except California) and the District of Columbia for 2000 provided to Centers for Disease Control and Prevention's National Center for Health Statistics were analyzed. Trends in maternal smoking were examined with data from birth certificates and other relevant sources. RESULTS: Smoking during pregnancy was reported for 12.2% of women who gave birth in 2000, down 37% from 1989 (19.5%), when this information was first collected on birth certificates. Throughout the 1990s, prenatal smoking rates were highest for older teenagers and women in their early 20s. Among population subgroups, the highest rates were reported for non-Hispanic white women who attended but did not complete high school. The incidence of LBW among singleton infants who were born to smokers was double that for nonsmokers. This relationship was observed in all age groups, for births to Hispanic and non-Hispanic white and black women, and within educational attainment subgroups. Even light smoking (<5 cigarettes daily) was associated with elevated rates of LBW. CONCLUSION: Although prenatal smoking may be underreported on the birth certificate, the trends and variations in smoking based on birth certificate data have been confirmed with data from other sources. Birth certificate data can be useful in monitoring prenatal smoking patterns. Changes in the birth certificate questions that are to be implemented beginning in 2003 will help to clarify the levels and changes in smoking behavior during pregnancy so that smoking cessation programs can be more effectively designed to meet the needs of the populations at risk. PMID- 12728135 TI - Contribution of excess weight gain during pregnancy and macrosomia to the cesarean delivery rate, 1990-2000. AB - OBJECTIVE: After declining for many years, cesarean delivery rates recently increased. To explore whether this increase is associated with excess weight gain during pregnancy, resulting in macrosomic infants who require cesarean delivery, we examined trends in excess weight gain, macrosomia, and cesarean delivery. METHODS: Analysis of 1990-2000 US Natality Files of birth certificate data were restricted to first birth, singleton infants of 37 to 42 weeks' gestation to avoid confounding by repeat cesarean delivery, complications of multigestational pregnancy, and preterm and postterm birth. Excess weight gain was defined according to current guidelines (41+ lb) and macrosomia as birth weight >4000 g. RESULTS: From 1990-2000, excess weight gain rose steadily from 18.6% to 24.2%. There was a 19.3% decline in macrosomic infants among women who gained excess weight compared with an 11.9% decline among women who gained 15-40 lb, although the absolute risk remained substantially greater among women who gained excess weight (eg, 14.2% vs 7.2%, in 2000). From 1990-1997, cesarean delivery declined by 20.2% among women who gained excess weight compared with 15.7% among women who gained 15 to 40 lb. After 1997, cesarean delivery increased in all weight gain categories, and absolute risks in 2000 were 25.8% for women who gained excess weight compared with 21.6% for women who gained 15-40 lb. Overall, women who gained excess weight accounted for 24.1% of cesarean deliveries in 1990 and 28.1% in 2000. CONCLUSIONS: Excess weight gain and macrosomia do not seem to be the primary factors that contribute to the recent increase in cesarean delivery because cesarean delivery rates have increased in all weight gain categories and macrosomia rates have decreased steadily from 1990-2000. Nonetheless, women who gain excess weight account for a growing proportion of cesarean deliveries because their relative numbers have grown. PMID- 12728136 TI - Missed opportunities for perinatal HIV prevention among HIV-exposed infants born 1996-2000, pediatric spectrum of HIV disease cohort. AB - OBJECTIVE: Despite dramatic reductions in perinatal human immunodeficiency virus (HIV) transmission in the United States, obstacles to perinatal HIV prevention that include lack of prenatal care; failure to test pregnant women for HIV before delivery; and lack of prenatal, intrapartum, or neonatal antiretroviral (ARV) use remain. The objective of this study was to describe trends in perinatal HIV prevention methods, perinatal transmission rates, and the contribution of missed opportunities for perinatal HIV prevention to perinatal HIV infection. METHODS: We analyzed data obtained from infant medical records on 4755 HIV-exposed singleton deliveries in 1996-2000, from 6 US sites that participate in the Centers for Disease Control and Prevention's Pediatric Spectrum of HIV Disease Project. HIV-exposed deliveries refer to deliveries in which the mother was known to have HIV infection during the pregnancy. RESULTS: Of the 4287 women with data on prenatal care, 92% had prenatal care. From 1996 to 2000, among the 3925 women with prenatal care, 92% had an HIV test before delivery; the use of prenatal zidovudine (ZDV) alone decreased from 71% to 9%, and the use of prenatal ZDV with other ARVs increased from 6% to 70%. Complete data on maternal and neonatal ARVs were available for 3284 deliveries. Perinatal HIV transmission was 3% in 1651 deliveries with prenatal ZDV in combination with other ARVs, intrapartum ZDV, and neonatal ZDV; 6% in 1111 deliveries with prenatal, intrapartum, and neonatal ZDV alone; 8% in 152 deliveries with intrapartum and neonatal ZDV alone; 14% of 73 deliveries with neonatal ZDV only started within 24 hours of birth; and 20% in 297 deliveries with no prenatal, intrapartum, and neonatal ARVs. Complete data on prenatal events were available in 328 HIV-infected and 3258 HIV-uninfected infants. A total of 56% of mothers of HIV-infected infants had missed opportunities for perinatal HIV prevention versus 16% of mothers of HIV uninfected infants. Forty-four percent of the infected infants were born to mothers who had prenatal care, a prenatal HIV diagnosis, and documented prenatal ARV therapy. Seventeen percent of women with reported illicit drug use had no prenatal care versus 3% of women with no reported drug use. In a multivariate analysis, maternal illicit drug use was significantly associated with lack of prenatal care. In a multivariate analysis, year of infant birth and the combination of lack of maternal HIV testing before delivery and lack of prenatal antiretroviral therapies were significantly associated with perinatal HIV transmission. CONCLUSIONS: Missed opportunities for perinatal HIV prevention contributed to more than half of the cases of HIV-infected infants. Prenatal care and HIV testing before delivery are major opportunities for perinatal HIV prevention. Illicit drug use was highly associated with lack of prenatal care, and lack of HIV testing before delivery was highly associated with perinatal HIV transmission. PMID- 12728137 TI - Hepatitis B surface antigen prevalence among pregnant women in urban areas: implications for testing, reporting, and preventing perinatal transmission. AB - OBJECTIVES: To estimate race/ethnicity-specific prevalence of hepatitis B surface antigen (HBsAg) in pregnant urban women and to evaluate factors associated with maternal HBsAg testing. METHODS: A multicenter, retrospective chart review was conducted of a racially/ethnically stratified random sample of maternal/infant charts of 10 523 women who gave birth to live infants during 1990-1993 in 4 urban areas in the United States. Data were collected on multiple variables, including demographic variables, HBsAg test dates and results, prenatal care type, and amount and source of payment. RESULTS: HBsAg prevalence among white non-Hispanics was 0.60% (95% confidence interval [CI]: 0.22-0.98), black non-Hispanics 0.97% (95% CI: 0.48-1.47), Hispanics 0.14% (95% CI: 0.01-0.26), and Asians 5.79% (95% CI: 4.42-7.16). HBsAg testing rates increased from 56.6% in 1990 to 78.2% in 1993. Factors associated with not being tested varied by urban area, but in the combined area model, they were having no or private prenatal care (odds ratios: 18.75 and 5.07, respectively) and being black (odds ratios: 2.08). Only 20.9% (95% CI: 19.1%-22.8%) of those not tested prenatally were tested at delivery. The expected number of infants born to HBsAg-positive study-area women was 3327 using study prevalence rates, compared with 1761 using national rates. CONCLUSIONS: To help ensure that all urban infants who are born to HBsAg-positive women receive appropriate prophylaxis, health officials in urban areas should use urban-area prevalence rates to ascertain completeness of reporting maternal HBsAg positivity. Needed steps to increase maternal HBsAg testing rates include ensuring that more pregnant women receive prenatal care, promoting testing by private providers, educating providers about testing in all racial and ethnic groups, and reminding providers to test at delivery those women not tested prenatally. PMID- 12728138 TI - Prevalence of breastfeeding in the United States: the 2001 National Immunization Survey. AB - OBJECTIVE: To address key gaps in the annual monitoring of breastfeeding prevalence in the United States, 3 breastfeeding questions concerning the initiation, duration, and exclusivity of breastfeeding were added to the rotating modules of the National Immunization Survey (NIS) beginning in the third quarter of 2001. The present study examines the current prevalence of breastfeeding in the United States using NIS data from this initial quarter. METHODS: The NIS is a random-digit-dialing survey of households with children aged 19 to 35 months, followed by a mail survey of the eligible children's vaccination providers to validate the child's vaccination information. In the third quarter of 2001, a randomly selected subset of households interviewed in the NIS (N = 896) were asked questions about breastfeeding. RESULTS: Almost two thirds (65.1%) of children had ever been breastfed. At 6 and 12 months, 27.0% and 12.3%, respectively, were receiving some breast milk. Non-Hispanic blacks had the lowest rates of breastfeeding initiation and continuation. Exclusive breastfeeding rates were low in the United States with only 7.9% at 6 months. CONCLUSIONS: Although breastfeeding initiation is near the national goal of 75%, breastfeeding continuation lags behind the national goals of 50% and 25% at 6 and 12 months, respectively. Strenuous public health efforts are needed to improve breastfeeding practices among blacks. PMID- 12728139 TI - Epidemiology of early hearing loss detection in Hawaii. AB - OBJECTIVE: Universal Newborn Hearing Screening began in 2 Honolulu hospitals in 1992, and by 1999, all 14 civilian birthing facilities in Hawaii were providing screening. Examination of 1998 Hawaii data indicated that approximately 13% of infants who did not pass initial hearing screening in the hospital did not return for the indicated follow-up. The purpose of this study was to determine the epidemiologic profile of infants who were born in 1999 and did not return for follow-up. METHODS: A population-based, cohort study of the hearing screening completion rates among the 13 civilian birthing facilities in Hawaii that provided data to the Department of Health was conducted. Analysis included a bivariate analysis of the demographic characteristics of infants who completed the screening/follow-up process compared with those who did not and logistic regression modeling to ascertain the demographic profile of infants at high risk for being lost to follow-up. RESULTS: Of 12 456 infants, hearing screening data could be linked to the birth certificate file, and a final disposition regarding completion of the screening/follow-up process was determined for 10 328 (83%). Less than 2% (n = 176) of the linked infants failed to complete the screening/follow-up procedures. Low birth weight and white infants and infants born to women who had not completed high school were approximately twice as likely not to complete the screening as were their normal birth weight or nonwhite counterparts. CONCLUSIONS: Failure to complete the hearing screening follow-up may be related to cultural differences that have been previously reported in other maternal and child health studies of the diverse populations in Hawaii. The results of this study will allow the Hawaii Newborn Hearing Screening Program to target its efforts and limited resources toward infants who are at higher risk of not completing the screening and who may need special attention to encourage their mothers to complete the screening process, and to move quickly with rescreening infants whose initial tests are positive so that infants are not lost to follow-up. PMID- 12728140 TI - Sleep environment and the risk of sudden infant death syndrome in an urban population: the Chicago Infant Mortality Study. AB - OBJECTIVE: To examine risk factors for sudden infant death syndrome (SIDS) with the goal of reducing SIDS mortality among blacks, which continues to affect this group at twice the rate of whites. METHODS: We analyzed data from a population based case-control study of 260 SIDS deaths that occurred in Chicago between 1993 and 1996 and an equal number of matched living controls to determine the association between SIDS and factors in the sleep environment and other variables related to infant care. RESULTS: The racial/ethnic composition of the study groups was 75.0% black; 13.1% Hispanic white; and 11.9% non-Hispanic white. Several factors related to the sleep environment during last sleep were associated with higher risk of SIDS: placement in the prone position (unadjusted odds ratio [OR]: 2.4; 95% confidence interval [CI]: 1.7-3.4), soft surface (OR: 5.1; 95% CI: 3.1-8.3), pillow use (OR: 2.5; 95% CI: 1.5-4.2), face and/or head covered with bedding (OR: 2.5; 95% CI: 1.3-4.6), bed sharing overall (OR: 2.7; 95% CI: 1.8-4.2), bed sharing with parent(s) alone (OR: 1.9; 95% CI: 1.2-3.1), and bed sharing in other combinations (OR: 5.4; 95% CI: 2.8-10.2). Pacifier use was associated with decreased risk (unadjusted OR: 0.3; 95% CI: 0.2-0.5), as was breastfeeding either ever (OR: 0.2; 95% CI: 0.1-0.3) or currently (OR: 0.2; 95% CI: 0.1-0.4). In a multivariate model, several factors remained significant: prone sleep position, soft surface, pillow use, bed sharing other than with parent(s) alone, and not using a pacifier. CONCLUSIONS: To lower further the SIDS rate among black and other racial/ethnic groups, prone sleeping, the use of soft bedding and pillows, and some types of bed sharing should be reduced. PMID- 12728141 TI - Maternal characteristics associated with vaccination of young children. AB - OBJECTIVE: Mothers can be instrumental in gaining access to vaccination services for their children. This study examines maternal characteristics associated with vaccination in US preschool children. METHODS: We analyzed data from 21 212 children aged 19 to 35 months in the National Immunization Survey. Bivariate and multivariate analyses were used to identify maternal characteristics associated with completion of all recommended vaccinations in these children. RESULTS: Factors most strongly associated with undervaccination included having mothers who were black; had less than a high school education; were divorced, separated, or widowed; had multiple children; were eligible for the Special Supplemental Nutrition Program for Women, Infants and Children (WIC) but not participating; or had incomes below 50% of the federal poverty level. CONCLUSION: Because most mothers play an important role in their children's vaccination, it is important to address maternal concerns and barriers when developing public health interventions for promoting childhood vaccinations. Encouraging eligible women and their children to participate in the WIC program and providing support and encouragement for immunization to mothers with multiple children may improve early childhood vaccination coverage. PMID- 12728142 TI - Trends in postneonatal mortality attributable to injury, United States, 1988 1998. AB - OBJECTIVE: Half of all postneonatal mortality (PNM; deaths among infants aged 28 364 days) in the United States is caused by potentially preventable causes such as sudden infant death syndrome, infections, and injuries. A detailed analysis of PNM attributable to injury has not been conducted and may provide useful data in prioritizing prevention strategies and targeting high-risk populations. METHODS: The authors used US infant death certificate data to analyze trends in PNM caused by injury during 1988-1998. Attending physicians, medical examiners, or coroners report cause of death on death certificates using a format specified by the World Health Organization and endorsed by the Centers for Disease Control and Prevention. The major causes of PNM by type of injury were evaluated, and trends were compared over time. Injury-related deaths per 100 000 live births were examined by race and region of residence. Rate ratios between black and white infants also were calculated. RESULTS: Among major causes of PNM during the study period, injury mortality declined the least (13.0% decline; from 29.6 to 25.7 per 100 000 live births). All types of unintentional injury deaths declined except for mechanical suffocation rates, which increased from 4.8 to 7.1. Homicide rates increased slightly (8.5%) from an 11-year low in 1988 and accounted for a greater proportion of all PNM caused by injury by 1998 (27.5% in 1998, 22.1% in 1988). Overall, PNM rates attributable to injury declined less among blacks (8.7%) than whites (13.6%) during the study period, and rates were on average 2.6 times higher among black infants (range: 2.4-3.0). Unintentional injury declined less among blacks (15.4%) than among whites (24.9%), in part because of an increase in motor vehicle crash-related mortality rates among black infants. Although black infants were more than 3 times as likely to be a victim of homicide than white infants (range: 3.0-4.4), increases in homicide rates were similar among black infants (9.9%) and white infants (10.6%) from 1988 through 1998. Racial disparities in PNM attributable to injury varied by region. PNM rates attributable to injury increased only among black infants residing in the Midwest (10.2%) and West (27.7%) as a result of increases in unintentional injury (ie, motor vehicle crash-related deaths in the West and mechanical suffocation in the Midwest) and homicide rates in these regions. Homicide rates increased among all infants regardless of race, except for infants residing in the Northeast. CONCLUSIONS: Overall PNM rates attributable to injury declined, yet rates of mechanical suffocation increased and large regional and racial disparities persisted. Death certificates have limited information to help explain the observed differences. Because injuries are frequently preventable, prevention strategies should encourage formation of infant and child death review teams to help identify community and system factors that may contribute to injury deaths. Health care providers can assist parents in providing a safe environment for infants by counseling on age-appropriate injury prevention as part of their anticipatory guidance and serving as child advocates. Additional studies should examine regional differences in death investigation practices, case ascertainment, and reporting of deaths attributed to intentional injuries. PMID- 12728143 TI - Maternal perceptions of weight status of children. AB - OBJECTIVE: We quantified maternal misclassification of child weight status and examined determinants associated with maternal perceptions of child weight status. METHODS: Data from the Third National Health and Nutrition Examination Survey (1988-1994) were used. The sample included 5500 children (aged 2-11 years) with maternal interview data. Maternal perceptions of children's weight status were compared with measured weights and statures from which body mass index (BMI; weight/stature2; kg/m2) percentiles and z scores were determined. Frequency analyses determined the percentages of mothers considering their child to be "overweight," "underweight," or "about the right weight." Multivariable logistic regression analyses determined predictors of maternal misclassification of overweight children (> or =95th BMI-for-age percentile) and those at risk for overweight (> or =85th to <95th BMI-for-age percentile). RESULTS: Nearly one third (32.1%) of mothers reported their overweight child as "about the right weight." Younger children and those with lower BMI-for-age z scores had significantly greater odds of maternal underclassification of child overweight status. For children at risk for overweight, 14.0% of mothers reported sons to be "overweight," whereas 29.0% considered daughters to be "overweight." Odds of maternal misclassification of at-risk children as "overweight" were significantly greater for daughters, older children, children with higher BMI-for-age z scores, and children whose mothers had a lower BMI. Race/ethnicity was not a significant predictor in either model. CONCLUSIONS: Nearly one third of mothers misclassify overweight children as being lower than their measured weight status. Mothers are more likely to identify daughters who are at risk of overweight as being "overweight" than they are sons. PMID- 12728144 TI - Attention-deficit/hyperactivity disorder in school-aged children: association with maternal mental health and use of health care resources. AB - OBJECTIVE: To investigate the association between the mental health status of mothers and attention-deficit/hyperactivity disorder (ADHD) in their school-aged children and to characterize the health care access and utilization of families affected by ADHD. METHODS: Survey logistic regression procedures were used to investigate the association between activity-limiting mental health conditions in mothers and ADHD in their school-aged children using 1998 National Health Interview Survey data. A total of 9529 mother-child dyads were included in the final analysis. RESULTS: The prevalence of ADHD among children aged 4 to 17 years was 6.3%. Survey logistic regression statistics revealed an association between an activity-limiting depression, anxiety, or emotional problem in mothers and ADHD in their children. This association persisted after controlling for the gender, age, and race of the child; household income (as a function of the 1997 poverty level); and type of family structure as reported by the mother (adjusted odds ratio [OR]: 4.2; 95% confidence interval [CI]: 2.2-8.1). Mothers of a child with ADHD were 13 times more likely to have consulted with a mental health professional about their child's health within the past year despite reporting an inability to afford prescription medications (OR: 3.3; 95% CI: 2.2-4.9) and mental health care (OR: 7.4; 95% CI: 4.6, 11.8) for the child. CONCLUSIONS: Maternal mental health is significantly associated with the presence of ADHD in school-aged children. This finding further supports a link between maternal mental health and behavioral outcomes in children. Health care utilization and access findings support a family-oriented system of care. PMID- 12728145 TI - Introduction to the methods used in the generation of the British Thoracic Society guidelines for the management of pleural diseases. PMID- 12728146 TI - BTS guidelines for the investigation of a unilateral pleural effusion in adults. PMID- 12728147 TI - BTS guidelines for the management of pleural infection. PMID- 12728148 TI - BTS guidelines for the management of malignant pleural effusions. PMID- 12728152 TI - Assessment of illness severity in community acquired pneumonia: a useful new prediction tool? PMID- 12728153 TI - Patients with COPD: do we fail them from beginning to end? PMID- 12728150 TI - BTS guidelines for the insertion of a chest drain. PMID- 12728154 TI - Usefulness of peak expiratory flow measurements: is it just a matter of instrument accuracy? PMID- 12728149 TI - BTS guidelines for the management of spontaneous pneumothorax. PMID- 12728155 TI - Defining community acquired pneumonia severity on presentation to hospital: an international derivation and validation study. AB - BACKGROUND: In the assessment of severity in community acquired pneumonia (CAP), the modified British Thoracic Society (mBTS) rule identifies patients with severe pneumonia but not patients who might be suitable for home management. A multicentre study was conducted to derive and validate a practical severity assessment model for stratifying adults hospitalised with CAP into different management groups. METHODS: Data from three prospective studies of CAP conducted in the UK, New Zealand, and the Netherlands were combined. A derivation cohort comprising 80% of the data was used to develop the model. Prognostic variables were identified using multiple logistic regression with 30 day mortality as the outcome measure. The final model was tested against the validation cohort. RESULTS: 1068 patients were studied (mean age 64 years, 51.5% male, 30 day mortality 9%). Age >/=65 years (OR 3.5, 95% CI 1.6 to 8.0) and albumin <30 g/dl (OR 4.7, 95% CI 2.5 to 8.7) were independently associated with mortality over and above the mBTS rule (OR 5.2, 95% CI 2.7 to 10). A six point score, one point for each of Confusion, Urea >7 mmol/l, Respiratory rate >/=30/min, low systolic(<90 mm Hg) or diastolic (/=65 years (CURB-65 score) based on information available at initial hospital assessment, enabled patients to be stratified according to increasing risk of mortality: score 0, 0.7%; score 1, 3.2%; score 2, 3%; score 3, 17%; score 4, 41.5% and score 5, 57%. The validation cohort confirmed a similar pattern. CONCLUSIONS: A simple six point score based on confusion, urea, respiratory rate, blood pressure, and age can be used to stratify patients with CAP into different management groups. PMID- 12728156 TI - Dexamethasone for treatment of patients mechanically ventilated for lower respiratory tract infection caused by respiratory syncytial virus. AB - BACKGROUND: A study was undertaken to evaluate the efficacy of dexamethasone in patients mechanically ventilated for lower respiratory infection caused by respiratory syncytial virus (RSV-LRTI). METHODS: In a multicentre randomised controlled trial patients were randomised to receive either intravenous dexamethasone (0.15 mg/kg 6 hourly for 48 hours) or placebo. End points were the duration of mechanical ventilation, length of stay (LOS) in the pediatric intensive care unit (PICU) and in hospital, and the duration of supplemental oxygen administration. RESULTS: Thirty seven patients received dexamethasone and 45 received placebo. There was no significant difference in any of the end points between the two groups. In a post hoc analysis patients were stratified into those with mild gas exchange anomalies (PaO(2)/FiO(2) >200 mm Hg and/or mean airway pressure 10 cm H(2)O, pneumonia group). In the 39 patients with bronchiolitis the duration of mechanical ventilation was 4.3 days shorter in the dexamethasone group than in the placebo group (4.9 v 9.2 days, 95% CI -7.8 to -0.8, p=0.02) and the duration of supplemental oxygen was 3.6 days shorter (7.7 v 11.3 days, 95% CI -8.0 to 0.1, p=0.048). No differences in end points were found in the pneumonia group. CONCLUSIONS: Dexamethasone had no beneficial effect in patients mechanically ventilated for RSV-LRTI but was found to have a beneficial effect in patients with bronchiolitis. PMID- 12728157 TI - Lung function and mortality in the United States: data from the First National Health and Nutrition Examination Survey follow up study. AB - BACKGROUND: A study was undertaken to define the risk of death among a national cohort of US adults both with and without lung disease. METHODS: Participants in the first National Health and Nutrition Examination Survey (NHANES I) followed for up to 22 years were studied. Subjects were classified using a modification of the Global Initiative for Chronic Obstructive Lung Disease criteria for chronic obstructive pulmonary disease (COPD) into the following mutually exclusive categories using the forced expiratory volume in 1 second (FEV(1)), forced vital capacity (FVC), FEV(1)/FVC ratio, and the presence of respiratory symptoms: severe COPD, moderate COPD, mild COPD, respiratory symptoms only, restrictive lung disease, and no lung disease. Proportional hazard models were developed that controlled for age, race, sex, education, smoking status, pack years of smoking, years since quitting smoking, and body mass index. RESULTS: A total of 1301 deaths occurred in the 5542 adults in the cohort. In the adjusted proportional hazards model the presence of severe or moderate COPD was associated with a higher risk of death (hazard ratios (HR) 2.7 and 1.6, 95% confidence intervals (CI) 2.1 to 3.5 and 1.4 to 2.0), as was restrictive lung disease (HR 1.7, 95% CI 1.4 to 2.0). CONCLUSIONS: The presence of both obstructive and restrictive lung disease is a significant predictor of earlier death in long term follow up. PMID- 12728158 TI - Self-management education for patients with chronic obstructive pulmonary disease: a systematic review. AB - BACKGROUND: The idea of self-management is to teach patients how to carry out the activities of daily living optimally in the face of their physiological impairment, and to prevent or decrease the severity of exacerbations by means of life style adaptation. In chronic obstructive pulmonary disease (COPD) the value of self-management education is not clear. This review was undertaken to clarify the effectiveness of self-management programmes in COPD. METHODS: A search was made of the Cochrane Airways Group trial registers, Medline, reference lists, and abstracts of medical conferences for controlled trials of self-management education in patients with COPD. Two reviewers independently assessed each paper for methodological quality and extracted the data. RESULTS: The reviewers included 12 articles describing eight randomised controlled trials and one controlled clinical trial in which self-management education was compared with usual care. The studies assessed a broad spectrum of outcome measures with different follow up times so meta-analysis could not be undertaken. Self management education had no effect on hospital admissions, emergency room visits, days lost from work, and lung function. Inconclusive results were observed on health related quality of life, COPD symptoms, and use of healthcare facilities such as doctor and nurse visits. Self-management education reduced the need for rescue medication and led to increased use of courses of oral steroids and antibiotics for respiratory symptoms. CONCLUSIONS: Insufficient data were obtained to make recommendations because of the wide variation in outcome measures used and other limitations to generalisations in the current published literature. Further research in this area is needed. PMID- 12728159 TI - Health outcomes following treatment for six months with once daily tiotropium compared with twice daily salmeterol in patients with COPD. AB - BACKGROUND: A study was undertaken to record exacerbations and health resource use in patients with COPD during 6 months of treatment with tiotropium, salmeterol, or matching placebos. METHODS: Patients with COPD were enrolled in two 6-month randomised, placebo controlled, double blind, double dummy studies of tiotropium 18 micro g once daily via HandiHaler or salmeterol 50 micro g twice daily via a metered dose inhaler. The two trials were combined for analysis of heath outcomes consisting of exacerbations, health resource use, dyspnoea (assessed by the transitional dyspnoea index, TDI), health related quality of life (assessed by St George's Respiratory Questionnaire, SGRQ), and spirometry. RESULTS: 1207 patients participated in the study (tiotropium 402, salmeterol 405, placebo 400). Compared with placebo, tiotropium but not salmeterol was associated with a significant delay in the time to onset of the first exacerbation. Fewer COPD exacerbations/patient year occurred in the tiotropium group (1.07) than in the placebo group (1.49, p<0.05); the salmeterol group (1.23 events/year) did not differ from placebo. The tiotropium group had 0.10 hospital admissions per patient year for COPD exacerbations compared with 0.17 for salmeterol and 0.15 for placebo (not statistically different). For all causes (respiratory and non respiratory) tiotropium, but not salmeterol, was associated with fewer hospital admissions while both groups had fewer days in hospital than the placebo group. The number of days during which patients were unable to perform their usual daily activities was lowest in the tiotropium group (tiotropium 8.3 (0.8), salmeterol 11.1 (0.8), placebo 10.9 (0.8), p<0.05). SGRQ total score improved by 4.2 (0.7), 2.8 (0.7) and 1.5 (0.7) units during the 6 month trial for the tiotropium, salmeterol and placebo groups, respectively (p<0.01 tiotropium v placebo). Compared with placebo, TDI focal score improved in both the tiotropium group (1.1 (0.3) units, p<0.001) and the salmeterol group (0.7 (0.3) units, p<0.05). Evaluation of morning pre-dose FEV(1), peak FEV(1) and mean FEV(1) (0-3 hours) showed that tiotropium was superior to salmeterol while both active drugs were more effective than placebo. CONCLUSIONS: Exacerbations of COPD and health resource usage were positively affected by daily treatment with tiotropium. With the exception of the number of hospital days associated with all causes, salmeterol twice daily resulted in no significant changes compared with placebo. Tiotropium also improved health related quality of life, dyspnoea, and lung function in patients with COPD. PMID- 12728160 TI - Influence of lung volume reduction surgery (LVRS) on health related quality of life in patients with chronic obstructive pulmonary disease. AB - BACKGROUND: The clinical value of LVRS has been questioned in the absence of trials comparing it with pulmonary rehabilitation, the prevailing standard of care in COPD. Patients with heterogeneous emphysema are more likely to benefit from volume reduction than those with homogeneous disease. Disease specific quality of life is a responsive interpretable outcome that enables health professionals to identify the magnitude of the effect of an intervention across several domains. METHODS: Non-smoking patients aged <75 years with severe COPD (FEV(1) <40% predicted, FEV(1)/FVC <0.7), hyperinflation, and evidence of heterogeneity were randomised to surgical or control groups after pulmonary rehabilitation and monitored at 3 month intervals for 12 months with no crossover between the groups. The primary outcome was disease specific quality of life as measured by the Chronic Respiratory Questionnaire (CRQ). Treatment failure was defined as death or functional decline (fall of 1 unit in any two domains of the CRQ). Secondary outcomes included pulmonary function and exercise capacity. RESULTS: LVRS resulted in significant between group differences in each domain of the CRQ at 12 months (change of 0.5 represents a small but important difference): dyspnoea 1.9 (95% confidence interval (CI) 1.3 to 2.6; p<0.0001); emotional function 1.5 (95% CI 0.9 to 2.1; p<0.0001); fatigue 2.0 (95% CI 1.4 to 2.6; p<0.0001); mastery 1.8 (95% CI 1.2 to 2.5; p<0.0001). In the control group one of 27 patients died and 16 experienced functional decline over 12 months. In the surgical group four of 28 patients died and three experienced functional decline (hazard ratio = 3.1 (95% CI 1.3 to 7.6; p=0.01). Between group improvements (p<0.05) in lung volumes, flow rates, and exercise were sustained at 12 months (RV -47% predicted (95% CI -71 to -23; p=0.0002); FEV(1) 0.3 l (95% CI 0.1 to 0. 5; p=0.0003); submaximal exercise 7.3 min (95% CI 3.9 to 10.8; p<0.0001); 6 minute walk 66 metres (95% CI 32 to 101; p=0.0002). CONCLUSIONS: In COPD patients with heterogeneous emphysema, LVRS resulted in important benefits in disease specific quality of life compared with medical management, which were sustained at 12 months after treatment. PMID- 12728161 TI - Inadequate peak expiratory flow meter characteristics detected by a computerised explosive decompression device. AB - BACKGROUND: Recent evidence suggests that the frequency response requirements for peak expiratory flow (PEF) meters are higher than was first thought and that the American Thoracic Society (ATS) waveforms to test PEF meters may not be adequate for the purpose. METHODS: The dynamic response of mini-Wright (MW), Vitalograph (V), TruZone (TZ), MultiSpiro (MS) and pneumotachograph (PT) flow meters was tested by delivering two differently shaped flow-time profiles from a computer controlled explosive decompression device fitted with a fast response solenoid valve. These profiles matched population 5th and 95th centiles for rise time from 10% to 90% of PEF and dwell time of flow above 90% PEF. Profiles were delivered five times with identical chamber pressure and solenoid aperture at PEF. Any difference in recorded PEF for the two profiles indicates a poor dynamic response. RESULTS: The absolute (% of mean) flow differences in l/min for the V, MW, and PT PEF meters were 25 (4.7), 20 (3.9), and 2 (0.3), respectively, at PEF approximately 500 l/min, and 25 (10.5), 20 (8.7) and 6 (3.0) at approximately 200 l/min. For TZ and MS meters at approximately 500 l/min the differences were 228 (36.1) and 257 (39.2), respectively, and at approximately 200 l/min they were 51 (23.9) and 1 (0.5). All the meters met ATS accuracy requirements when tested with their waveforms. CONCLUSIONS: An improved method for testing the dynamic response of flow meters detects marked overshoot (underdamping) of TZ and MS responses not identified by the 26 ATS waveforms. This error could cause patient misclassification when using such meters with asthma guidelines. PMID- 12728163 TI - Allelic association and functional studies of promoter polymorphism in the leukotriene C4 synthase gene (LTC4S) in asthma. AB - BACKGROUND: LTC4 synthase is essential for the production of cysteinyl leukotrienes (Cys-LT), critical mediators in asthma. We have identified a novel promoter polymorphism at position -1072 (G/A) and a -444 (A/C) polymorphism has previously been reported. The role of these polymorphisms in the genetic susceptibility to asthma was examined. METHODS: To test for genetic association with asthma phenotypes, 341 white families (two asthmatic siblings) and 184 non asthmatic control subjects were genotyped. Genetic association was assessed using case control and transmission disequilibrium test (TDT) analyses. LTC4S promoter luciferase constructs and transiently transfected human HeLa and KU812F cells were generated to determine the functional role of these polymorphisms on basal transcription. RESULTS: No associations were observed in case control analyses ( 1072 A, q=0.09; -444 C, q=0.29); the TDT identified a borderline association between the -444 C allele and bronchial responsiveness to methacholine (p=0.065). Asthmatic children with the -444 C allele had a lower mean basal forced expiratory volume in 1 second (97.4 v 92.7% predicted, p=0.005). LTC4S promoter luciferase analyses provided no evidence for a functional role of either polymorphism in determining basal transcription. CONCLUSION: This study does not support a role for these polymorphisms in genetic susceptibility to asthma but provides evidence to suggest a role in determining lung function parameters. PMID- 12728165 TI - Increased concentrations of human beta-defensins in plasma and bronchoalveolar lavage fluid of patients with diffuse panbronchiolitis. AB - BACKGROUND: Human beta-defensin (HBD)-1 and -2 are antimicrobial peptides present in the respiratory tract. Recent reports have indicated reduced activity of beta defensins in cystic fibrosis, suggesting that beta-defensins may play an important role in the pathological process of chronic respiratory tract infection. Diffuse panbronchiolitis (DPB) is a progressive disease characterised by frequent episodes of superimposed infection, typically caused by Pseudomonas aeruginosa. The aim of this study was to elucidate the role of these antimicrobial peptides in this disease. METHODS: The concentrations of HBD-1 and HBD-2 in plasma and bronchoalveolar lavage (BAL) fluid from 33 patients with DPB and 30 normal adults were measured by radioimmunoassay. Localisation of HBD-2 was investigated immunohistochemically in an open lung biopsy specimen obtained from a patient with DPB. RESULTS: High concentrations of HBD-1 and HBD-2 were noted in BAL fluid from DPB patients. Increased plasma concentrations of HBD-2, but not HBD-1, were found in patients with DPB compared with control subjects. In patients with DPB the HBD-2 concentration in BAL fluid correlated significantly with the numbers of cells recovered from the BAL fluid (total cells, neutrophils, and lymphocytes) and with the BAL fluid concentration of IL-1beta. Synthetic HBD 2, but not HBD-1, had dose dependent bactericidal activity against P aeruginosa. Treatment of 14 patients with macrolides significantly reduced BAL fluid concentrations of HBD-2 but not HBD-1 or plasma concentrations of HBD-1 and HBD 2. Immunohistochemistry of lung tissue showed localisation of HBD-2 in the epithelia of the distal bronchioles. CONCLUSIONS: These results indicate that beta-defensins, particularly HBD-2, participate in antimicrobial defence in the respiratory tract in DPB, and that the BAL fluid concentration of HBD-2 may be a useful marker of airway inflammation in patients with DPB. PMID- 12728166 TI - Episodic viral wheeze in preschool children: effect of topical nasal corticosteroid prophylaxis. AB - BACKGROUND: The effect of prophylactic nasal corticosteroids on wheezing episodes associated with colds was investigated in a 12 week parallel group, double blind, randomised controlled trial in preschool children. METHODS: Data were collected from 50 children aged 12-54 months with a history of at least three episodes of wheeze associated with colds over the previous winter, but few or no interval symptoms; 24 were given one dose of fluticasone aqueous nasal spray (50 micro g) into each nostril twice daily and 26 received an indistinguishable placebo spray. Episodes of lower respiratory illness occurring within 2 days of the onset of a cold were identified from daily symptom diaries. The main outcome was nocturnal symptom score during the first 7 days of an episode. RESULTS: The groups were well balanced on entry except that the treatment group had a history of more prolonged episodes. During the trial there was no significant difference in the number of episodes in the treatment and control groups (27 and 37, respectively), in the severity of nocturnal symptoms (mean score 1.33 and 1.22, respectively, confidence interval of difference -0.24 to +0.47) or in daytime symptoms, activity or total scores during episodes. Compliance was estimated to be over 50% in 43 of the children. CONCLUSIONS: Nasal corticosteroid treatment does not prevent acute wheezy episodes associated with upper respiratory infections (common colds) in preschool children. PMID- 12728167 TI - Health effects associated with smokeless tobacco: a systematic review. AB - BACKGROUND: It is believed that health risks associated with smokeless tobacco (ST) use are lower than those with cigarette smoking. A systematic review was therefore carried out to summarise these risks. METHODS: Several electronic databases were searched, supplemented by screening reference lists, smoking related websites, and contacting experts. Analytical observational studies of ST use (cohorts, case-control, cross sectional studies) with a sample size of >/=500 were included if they reported on one or more of the following outcomes (all cause mortality, oral and pharyngeal cancers, other cancers, cardiovascular diseases, dental diseases, pregnancy outcomes, surgical outcomes). Data extraction covered control of confounding, selection of cases and controls, sample size, clear definitions and measurements of the health outcome, and ST use. Selection, extraction and quality assessments were carried out by one or two independent reviewers. RESULTS: A narrative review was carried out. Many of the studies lacked sufficient power to estimate precise risks, mainly due to the small number of ST users. Studies were often not designed to investigate ST use, and many also had major methodological limitations including poor control for cigarette smoking and imprecise measurements of exposure. Studies in India showed a substantial risk of oral or oropharyngeal cancers associated with chewing betel quid and tobacco. Studies from other regions and of other cancer types were not consistent. Few studies have adequately considered the non-cancer health effects of ST use. CONCLUSIONS: Chewing betel quid and tobacco is associated with a substantial risk of oral cancers in India. Most recent studies from the US and Scandinavia are not statistically significant, but moderate positive associations cannot be ruled out due to lack of power. Further rigorous studies with adequate sample sizes are required, especially for cardiovascular disease. PMID- 12728169 TI - Preliminary findings of quorum signal molecules in clinically stable lung allograft recipients. AB - BACKGROUND: Infection with bacteria such as Pseudomonas is common in lung allograft recipients, particularly during chronic rejection. Analysis of sputum samples from patients with cystic fibrosis infected with Pseudomonas aeruginosa or Burkholderia cepacia has indicated the presence of bacterial N-acylhomoserine lactones (AHLs) quorum sensing signalling molecules. AHLs not only control the expression of bacterial virulence genes but are also involved in stimulating the maturation of antibiotic resistant biofilms and host chemokine release. It was hypothesised that AHLs may be detected even in clinically stable lung transplant recipients free of clinical infection or rejection. METHODS: Three 60 ml samples of bronchoalveolar lavage (BAL) fluid were taken from nine stable lung transplant recipients 3-12 months after transplantation. Detection of AHLs was carried out on dichloromethane extracted supernatants using the bioluminescence based AHL reporter plasmid pSB1075. This responds to the presence of AHLs with long acyl chains (C10-C14), generating light. Synthetic AHLs were included as positive controls. RESULTS: Five of the nine BAL fluid supernatants exhibited AHL activity, suggesting the presence of AHLs with long N-acyl chains. There was no correlation between the levels of AHLs detected or their absence and BAL fluid microbiology or diagnosis before transplantation. CONCLUSIONS: This is the first evidence for the presence of AHL quorum sensing signals in human lung allograft recipients, even in subjects with no rejection or apparent infection. Further longitudinal follow up of these preliminary findings is required to elucidate potential links with infection, rejection, and allograft deterioration. PMID- 12728171 TI - Lung cancer 5: state of the art radiotherapy for lung cancer. AB - Radiotherapy has a key role in curative and palliative treatments of patients with lung cancer. Important advances are described in the technique of treatment delivery and its integration with chemotherapy. PMID- 12728175 TI - Upper airways abnormalities and tracheal problems in Morquio's disease. AB - Morquio's disease is a metabolic disorder that can cause various respiratory abnormalities. Patients who live into adulthood are likely to develop upper airway problems and respiratory failure. With advances in home ventilation, these patients are increasingly likely to be referred to specialist respiratory units. We describe our experiences with two such patients. PMID- 12728177 TI - Dysfunctional breathing in COPD. PMID- 12728173 TI - Chronic obstructive pulmonary disease 8: non-pharmacological management of COPD. AB - The role of smoking cessation and the use of measures to reduce the disability associated with COPD are reviewed. The political profile of patients with COPD is increasing as patient support groups develop the confidence to campaign for better services. PMID- 12728176 TI - Diaphragm plication following phrenic nerve injury. PMID- 12728178 TI - Exertional haemoptysis: LAM and TSC. PMID- 12728179 TI - Lung function in preschool children. PMID- 12728180 TI - Occupational asthma evaluation. PMID- 12728181 TI - Whole-brain CT perfusion measurement of perfused cerebral blood volume in acute ischemic stroke: probability curve for regional infarction. AB - PURPOSE: To determine the probability curve for regional cerebral infarction as a function of percentage normalized perfused cerebral blood volume (pCBV) in patients with acute ischemic stroke. MATERIALS AND METHODS: The authors retrospectively analyzed whole-brain computed tomographic (CT) perfusion scans from 28 patients with acute stroke (<6 hours) due to major arterial occlusion, without intracranial hemorrhage. Each patient had a positive follow-up CT scan 1 4 days later, without interval thrombolysis. Normalized pCBV, expressed as a percentage of contralateral normal brain pCBV, was determined in the core infarction and in regions just inside and outside the boundary between infarcted and noninfarcted brain. These regions were dichotomized into infarcted (core and inner band) and noninfarcted (outer band) categories. Logistic regression analysis was then used to create a reference curve of probability of infarction as a function of percentage normalized pCBV. RESULTS: Normalized pCBV values in the core, inner band, and outer band were 24.5% +/- 2.3, 36.3% +/- 2.4, and 72.1% +/- 2.4, with corresponding probabilities of infarction of .99, .96, and .11. The normalized pCBV at which the probability of survival reached .5 was 58.0% +/- 0.5. Sensitivity, specificity, and accuracy of the reference probability curve were 90.5% (209 of 231), 89.5% (212 of 237), and 90.0% (421 of 468), respectively. Negative and positive predictive values were 90.6% (212 of 234) and 89.3% (209 of 234), respectively. R2 was 0.73, and differences in perfusion between core and inner and outer bands were highly significant (P <.0001). CONCLUSION: A probability of infarction curve can help predict the likelihood of infarction as a function of percentage normalized pCBV. PMID- 12728182 TI - Survey of radiology residents: breast imaging training and attitudes. AB - PURPOSE: To investigate the training and attitudes of residents regarding breast imaging. MATERIALS AND METHODS: A telephone survey was conducted with 201 4th year residents (postgraduate medical school year 5) and 10 3rd-year residents (postgraduate medical school year 4) at 211 accredited radiology residencies in the United States and Canada. Survey topics included organization of the breast imaging section, residents' role in the section, clinical practice protocols of the training institution, residents' personal thoughts about breast imaging, and their interest in performing breast imaging in the future. RESULTS: Of 211 programs, 203 (96%) had dedicated breast imaging rotations; 196 (93%) rotations were 8 weeks or longer; 153 (73%), 12 weeks or longer. Residents dictated reports in 199 (94%) programs. Residents performed real-time ultrasonography (US) in 186 (88%) programs, needle localization in 199 (94%), US-guided biopsy in 174 (82%), and stereotactically guided biopsy in 181 (86%). One hundred eighty-four (87%) residents rated interpretation of mammograms more stressful than they did that of other images, and 137 (65%) believed mammograms should be interpreted by subspecialists. One hundred thirty-five (64%) residents would not consider a fellowship in breast imaging if offered, and 133 (63%) would not want to spend 25% or more of their time in clinical practice on interpretation of mammograms. The most common reasons given for not considering a fellowship or interpretation of mammograms were that breast imaging was not an interesting field, that they feared lawsuits, and that it was too stressful. Fellowships were offered at 53 programs, and at 46 programs, a total of 63 fellows were recruited. CONCLUSION: Residency training in breast imaging has improved in terms of time and curriculum. However, a majority of the residents would not consider a fellowship and did not want to interpret mammograms in their future practices. PMID- 12728183 TI - Simple ovarian cysts in postmenopausal patients with breast carcinoma treated with tamoxifen: long-term follow-up. AB - PURPOSE: To assess the long-term natural history of simple ovarian cysts diagnosed in postmenopausal patients with breast carcinoma treated with tamoxifen citrate. MATERIALS AND METHODS: Of 332 postmenopausal women with breast cancer who were treated with tamoxifen, 32 (9.6%) had simple ovarian cysts. Long-term follow-up transvaginal ultrasonography (US) was performed in patients who had these simple ovarian cysts, and serum CA 125 samples were taken. Standard linear regression analysis with repeated measurements with irregular time points with the mixed-effects model was used to correlate cyst size at transvaginal US with the time elapsed since the diagnosis of ovarian cysts. Statistical analysis was performed by using the t test for regression slope. RESULTS: There was a significant decrease in cyst size over time (P =.017). Three (9%) of the 32 patients underwent surgery. Histologic evaluation of the removed ovaries revealed simple ovarian cyst, well-differentiated ovarian carcinoma, and metastatic adenocarcinoma. The remaining 29 (91%) continued with regular follow-up examinations only. In 11 (34%) of the 32 patients there were no changes in cyst size over time. In nine patients (28%) additional cysts appeared. Cysts disappeared, increased in size, disappeared and reappeared, or decreased in size in four (12%) patients each. Serum CA 125 levels were within the normal range. CONCLUSION: In postmenopausal patients with breast carcinoma who were treated with tamoxifen, long-term follow-up US of simple ovarian cysts demonstrates a significant decrease in cyst size over time. PMID- 12728184 TI - Coronary artery calcium: accuracy and reproducibility of measurements with multi detector row CT--assessment of effects of different thresholds and quantification methods. AB - PURPOSE: To evaluate the effects of different thresholds and quantification methods on the accuracy and reproducibility of coronary calcium measurements with multi-detector row computed tomography (CT). MATERIALS AND METHODS: A cardiac CT phantom containing predetermined calcified cylinders was scanned. Calcium volume and mass were measured at various threshold values ranging from 80 to 230 HU. In 32 patients, two consecutive CT scans were obtained, and the coronary artery calcium score, volume, and mass were measured by one observer at 130- and 90-HU thresholds. Correlation analysis and analysis of variance were performed to evaluate the measurement errors in the phantom study and the interscan variability in the clinical study. RESULTS: In the phantom, mass measurement error varied with threshold and calcium density (P <.01). Mass error was strongly correlated with volume error (r = 0.91, P <.01) but with a much smaller range. In the clinical study, interscan variability of mass measurements was significantly lower than that with other measurement methods for both patients and individual vessels. For the patients, the mean interscan variability of calcium score, volume, and mass at the 130-HU threshold was 20.4%, 13.9%, and 9.3%, respectively. For all methods, interscan variability was not significantly different between the 130- and 90-HU thresholds (P >.05). CONCLUSION: The mass measurement is more accurate, less variable, and more reproducible in coronary calcium quantification than are measurements with other algorithms. Accurate quantification of calcium in each calcified plaque may require that the threshold be set individually, depending on the calcium density. PMID- 12728185 TI - Differentiation of white, mixed, and red thrombi: value of CT in estimation of the prognosis of thrombolysis phantom study. AB - PURPOSE: To test with serial computed tomographic (CT) scans whether white, mixed, and red thrombi could be differentiated according to their CT attenuation. MATERIALS AND METHODS: Platelet-enriched plasma and whole blood were mixed to produce samples with hematocrit levels of 0, 0.005, 0.03, 0.15, and 0.35. A thrombin solution was added, and after 2 hours the retracted clots were transferred into polyethylene tubes with a length of 4 cm and an inner diameter of 3 mm. Ten probes of each sample were placed into a plastic box filled with a solution of gelatin, gadopentetate dimeglumine, and distilled water. Ten tubes filled with gelatin served as control. With this phantom, the CT numbers of white, mixed, and red thrombi were measured over 144 hours. CT was performed with a multisection scanner and a collimation of 0.5 mm. Statistical analyses were performed for differences between groups and over time. RESULTS: The CT numbers of white, mixed, and red thrombi differed significantly (P <.05) for most time measurements, except for white and mixed thrombi, which had a low hematocrit level at 24 and 144 hours (P >.05). CONCLUSION: With CT it appears feasible to differentiate thrombi according to their hematocrit level and thus estimate the effectiveness of thrombolysis. PMID- 12728186 TI - Inflammatory myofibroblastic tumor of the hepatobiliary system: report of MR imaging appearance in four patients. AB - The purpose of this report is to describe the magnetic resonance (MR) imaging features of hepatic inflammatory myofibroblastic tumor (IMT). Two observers retrospectively analyzed images from six MR imaging examinations in four consecutive patients. Tumor location, size, extent, morphology, and number; signal intensity on nonenhanced images; enhancement pattern on serial gadolinium enhanced images; duct wall thickness; and presence of biliary dilatation, vascular invasion, and lymphadenopathy were assessed. MR imaging findings were compared with histologic findings. On MR images, hepatic IMT manifested as a mass like lesion in two patients and an area of periportal soft-tissue infiltration in three; one patient had both patterns. Because periportal infiltration is a common feature in both IMT and other malignant tumors of the porta hepatis, histologic examination should be considered before diagnosis and treatment. PMID- 12728187 TI - Dyslexia (specific reading disability). PMID- 12728188 TI - Postpartum depression: what pediatricians need to know. PMID- 12728189 TI - Counseling the adolescent about contraception. PMID- 12728190 TI - Index of suspicion. PMID- 12728192 TI - Brain damage results in down-regulation of N-acetylaspartate as a neuronal osmolyte. AB - N-acetyl-L-aspartate (NAA) is present in the vertebrate brain, where its concentration is one of the highest of all free amino acids. Although NAA is synthesized and stored primarily in neurons, it is not hydrolyzed in these cells. However, after its regulated release into extracellular fluid, neuronal NAA is hydrolyzed by amidohydrolase II that is present in oligodendrocytes. About 30% of neurons do not contain appreciable amounts of NAA, but its prominence in 1H nuclear magnetic resonance spectroscopic (MRS) studies has led to its wide use as a neuronal marker in diagnostic human medicine as both an indicator of brain pathology, and of disease progression in a variety of central nervous system (CNS) diseases. Loss of NAA has been interpreted as indicating either loss of neurons, or loss of neuron viability. In this investigation, the upregulation of NAA in early stages of construction of the CNS, and its downregulation in experimentally induced damage models of the CNS is reported. The results of this study indicate that the buildup of NAA is not required for viability of neurons in monocellular cultures, and that NAA is lost from multicellular cultured brain slice explants that contain viable neurons. Thus, loss of NAA does not necessarily indicate either loss of neurons or their function. The NAA system, when present in the brain, appears to reflect a high degree of cellular integration, and therefore may be a unique metabolic construct of the intact vertebrate brain. PMID- 12728193 TI - Decrease in glial glutamate transporter variants and excitatory amino acid receptor down-regulation in a murine model of ALS-PDC. AB - Glutamate transporter proteins appear crucial to controlling levels of glutamate in the central nervous system (CNS). Abnormal and/or decreased levels of various transporters have been observed in amyotrophic lateral sclerosis (ALS) and Alzheimer's disease (AD) and in other neurological disorders. We have assessed glutamate transporter (GLT-1/EAAT2) levels in mice fed washed cycad flour containing a suspected neurotoxin that induces features resembling the Guamanian disorder, ALS-PDC. Down-regulation of glutamate transporter subtypes was detected by immunohistology using antibodies specific for two glial glutamate transporter splice variants (GLT-1alpha and GLT-1B). Immunohistology showed a "patchy" loss of antibody label with the patches centered on blood vessels. Computer densitometry showed significantly decreased GLT-1alpha levels in the spinal cord and primary somatosensory cortex of cycad-fed mice. GLT-1B levels were significantly decreased in the spinal cord, in the motor, somatosensory, and piriform cortices, and in the striatum. Western blots showed a 40% decrease in frontal motor cortex and lumbar spinal cord of cycad-fed mice that appeared to be phosphorylation-dependent. Receptor-binding assays showed decreased NMDA and AMPA receptor levels and increased GABAA receptor levels in cycad-fed mice cortex. These receptor data are consistent with an increased level of extracellular glutamate. The generalized decrease in GLT-1, decreased excitatory amino acid receptor levels, and increased GABAA receptor levels may reflect an early glutamate-mediated excitotoxicity following cycad exposure. Deciphering the series of events leading to neurodegeneration in cycad-fed animals may provide clues leading to therapeutic approaches to halt the early stages of disease progression. PMID- 12728197 TI - Perspectives: parity--prelude to a fifth cycle of reform. AB - BACKGROUND: Based on 2000 Carl Taube Lecture at the NIMH Mental Health Economics Meeting. AIMS OF THE STUDY: This perspective article examines the relationship between a policy of parity in financing mental health services and the future of reform in service delivery. METHODS: Applying theories of static and dynamic efficiency to an understanding of parity and the evolution of mental health services, drawing upon Burton Weisbrod s concept of the health care quadrilemma . RESULTS: Each of four cycles of reform in mental health services have contended with issues of static and dynamic efficiency. Each cycle was associated with static efficiency in the management and financing of services, and each was associated with a set of new treatment technologies intended to improve dynamic efficiency. Each reform proved ultimately unsuccessful primarily because of the failure of the treatment technologies to prevent future patient chronicity or to achieve sustained recovery. Recent advances in treatment technology and management of care can permit an unprecedented level of efficiency consistent with a policy of improved access to mainstream health and social welfare resources, including insurance coverage. This policy of so-called financing parity can improve current mental health service delivery, but it may also portend a future fifth cycle of reform. If new technologies continue to advance as full technologies - simple to deliver and producing true recovery - and mainstream resources are made available, then the specialty mental health services may contract dramatically in favor of effective care and treatment of mental illness in primary care and other mainstream settings. DISCUSSION: Predicting the future of health care is speculative, but it may be easier using the Weisbrod formulation to understand the process of mental health reform. Over reliance on administrative techniques for building static efficiency and false optimism about dynamic efficiency from new technology have stymied previous reforms. All the same, a fifth cycle of reform could succeed, if the right conditions are met and mainstream resources are available. PMID- 12728194 TI - Homocysteine and folate deficiency sensitize oligodendrocytes to the cell death promoting effects of a presenilin-1 mutation and amyloid beta-peptide. AB - Although damage to white matter occurs in the brains of patients with Alzheimer's disease (AD), the underlying mechanisms are unknown. Recent findings suggest that individuals with elevated levels of homocysteine are at increased risk of AD. Here we show that oligodendrocytes from mice expressing a mutant form of presenilin-1 (PS1) that causes familial AD exhibit increased sensitivity to death induced by homocysteine compared to oligodendrocytes from wild-type control mice. Homocysteine also sensitized oligodendrocytes to the cytotoxicity of amyloid beta peptide. Folate deficiency, which is known to result in elevated levels of homocysteine in vivo, also sensitized oligodendrocytes to the cell-death promoting actions of mutant PS1 and amyloid beta-peptide. Inhibitors of poly (ADP ribose) polymerase and p53 protected oligodendrocytes against cell death induced by homocysteine and amyloid beta-peptide, consistent with a role for a DNA-damage response in the cell death process. These findings demonstrate an adverse effect of homocysteine on oligodendrocytes, and suggest roles for homocysteine and folate deficiency in the white matter damage in AD and related neurodegenerative disorders. PMID- 12728196 TI - Unit cost of counseling and patients length of stay in a residential drug treatment setting. AB - BACKGROUND: Many published reports on cost of counseling give a fixed cost per hour of service. These estimates may be flawed. AIMS OF THE STUDY: The purpose of this study is to show, by way of an example, how cost of an hour of counseling depends on the nature of the patient, in general, and length of the patient s stay, in particular. Even though the health care professional provides the same hour of work, the cost of the hour is different for short-stay and long-stay patients. METHODS: We identified 5-short and 5 long stay patients in a residential treatment program. For each group, we asked the counselors to review the medical records and measure the patients utilization of various service units. We estimated the cost of a unit of service by dividing cost of an average patient by the program utilization of short and long-stay patients. RESULTS: The cost of an hour of counseling for long stay patients was 2/3 less than the cost of short-stay patients. Similar large changes in unit cost of treatment were observed for cost of group counseling or other components of substance abuse treatment. DISCUSSION: Our data was limited to one case study and may not indicate similar patterns in other treatment programs. The paper suggests that methods of studying cost of treatment should be adjusted to reflect case mix of patients and their expected length of stay. IMPLICATIONS FOR HEALTH POLICIES: Our analysis shows that higher rates should be set for patients at risk for short stays; conversely lower rates should be set for patients likely to complete treatment. Without adjusting the rate for the case mix of patients, health care institutions have an incentive to avoid the difficult cases and concentrate on long stay cases. IMPLICATIONS FOR FURTHER RESEARCH: A number of instruments that measure severity of illness or difficulty of treatment can be used to anticipate patients length of stay. Then the rate for units of treatment can be set based on patients expected length of stay. This paper presents a questionnaire that can be used to collect cost data and estimate cost per unit of treatment adjusted for expected length of stay. PMID- 12728198 TI - Cost-outcome of anxiety treatment intervention in primary care in Hungary. AB - AIM OF THE STUDY: The purpose of this paper is to estimate the changes in health utilization and indirect costs of anxiety and affective disorders in primary care patients after initiation of mental health treatment. METHOD: This study was conducted in 12 general practices for the primary care of adult populations in Budapest, Hungary. Among 2,000 eligible patients aged 18 to 64 years, 1,815 gave written informed consent to participate in the study. The Hungarian version of the Diagnostic Interview Schedule (DIS) for anxiety and mood disorders was used to generate psychiatric diagnoses. For all patients, health care utilization data for the previous 12 months was collected including number of visits, specialist consultations, days spent in hospital, sick days in the last year and prescribed medication. Among the first 1,000 attenders, 151 patients were given DIS/DSM-III R diagnoses of current anxiety and/or mood disorder or uncomplicated bereavement. Fifty-one patients who agreed to psychiatric treatment were assigned to the treatment group. After the first 1,000 participants, 75 patients were given DIS diagnoses and were considered as controls. In the treatment group, five psychiatrists administered treatment on an outpatient basis for one year. Patients in the control group received as-usual treatment from their primary care physicians. After one year, health care utilization data for the study period was collected. For the purposes of this study, the direct costs considered were limited to health care expenses and the indirect costs were limited to lost workdays. Statistical significance was calculated using a paired-samples T-test procedure comparing the means of two variables for a simple group. RESULTS: In the treatment group, the total cost of prescription drugs increased sharply due to psychiatric drug treatment, thus increasing the direct overall costs of care. In this same group the cost of non-psychiatric drugs showed a 37% decrease, suggesting that a reduction in general medical treatment partially offset the costs of anxiety and depression treatment. The number of hospital days showed marked decrease in the treatment group and a slight, insignificant increase in the control group. Absenteeism fell sharply in the treatment group (-56%) and in the group of patients who received psychiatric treatment elsewhere (-62%). In the control group, there was a large upturn (+182%) in the number of days spent on sick leave. DISCUSSION: Among primary care patients diagnosed with anxiety or affective disorders, psychiatric treatment led to higher direct costs, but this was offset by a decline in indirect costs due to reduced absenteeism compared with ordinary primary care. LIMITATIONS: Patients were not assigned randomly to the different groups because of ethical concerns. There were also significant differences in the baseline characteristics of the groups. Differences in the severity of illness and reasons not attributable to treatment effects may play a role in the change in the rate of service use. IMPLICATIONS FOR HEALTH POLICY: Limiting anxiety patients access to psychiatric treatment causes an increase in absenteeism, thus resulting in higher indirect costs. PMID- 12728200 TI - [Scientific research and scientific journals] AB - Not available PMID- 12728199 TI - Incentives in financing mental health care in Austria. AB - BACKGROUND: In Austria, financing health care -and even more so mental health care- is characterized by a mix of federal and provincial responsibilities, lack of uniformity in service provision and service providers, and diverse funding arrangements. The division between financing structures for health care and social care makes the situation even more complex. This state of affairs results in various, partly counterproductive and sometimes paradoxical financial incentives and disincentives for the providers, recipients and financiers of mental health services. In several provinces of Austria, recent reform plans in mental health care have focused strongly on establishing community-based and patient-oriented mental health care. One of the main challenges in implementing this new policy is the re-allocation of resources. AIMS OF THE STUDY: The authors hypothesize that the existing structure of mental health care financing, with its incentives and disincentives, constitutes an obstacle to patient-oriented community-based mental health care. Analyzing the characteristics of the overall mental health care financing system in one Austrian province, Lower Austria, will provide a better understanding of actor-relationships and inherent incentives and highlight implications for the process of deinstitutionalization. METHOD: The authors used an analytical framework based on the principal-agent theory, empirical evidence, and information on financial, organizational and legal structures to identify the characteristics of actor-relationships and the position of single actors within the system. RESULTS: The article shows how incentives are linked to existing constellations of actors involved in mental health care financing and identifies significant power relations. As a consequence, incentives and disincentives within the financing system result in hospital- centered and supply-oriented mental health care in Lower Austria. DISCUSSION: The current system of financing mental health care provides an obstacle to the provision of patient-oriented and community-based mental care. This is due to existing constellations and power relations among the actors where, most importantly, patients are the weakest party in the patient-payer provider triangle. Balancing power relations will be a significant prerequisite for alternative financing systems. IMPLICATIONS FOR HEALTH POLICIES AND FURTHER RESEARCH: If a community and needs-based mental health care system is to be established in Austria, the financing structures have to be changed accordingly. Applying a principal-agent framework is useful for identifying key aspects in mental health care financing in relation to the provision of services. Further research is needed to help develop alternative financing mechanisms that support community-based and patient-oriented mental health care systems. PMID- 12728201 TI - [Benefit/risk ratio of antibiotic therapy] AB - This publication evaluates critically the benefit/risk profile of several antibiotics currently at our disposal. The considered antibiotics are divided into pharmacological classes, because generally the drugs of the same class share the same adverse events. Moreover, the high therapeutic profile of the antibiotics puts them at the top of the safest drugs. Therefore the choice of an antibiotic is based above all on the evaluation of the patient and of the pathology to be treated, in terms of severity and possibility of achieving a response to treatment. An accurate anamnesis, the identification of the correct dosage and of the therapy duration minimise the potential risks of the chosen treatment. Detailed knowledge of the safety profile of these drugs is a further element in order for the antibiotic to perform at its best PMID- 12728202 TI - [Mortality among drug addicts in Bologna - A cohort study (1977-1997)] AB - Our study concerns the life status at the 31st december1997 of 1214 DA (drug addicts, mostly heroin addicted), enrolled by a SerT (Servizio per Tossicodipendenti, DA Healthcare Unit) in Bologna over the 1977-1996 period. All subjects were categorised by their HIV status (positive, negative or unknown), life status (alive or dead) and cause of death. Data were analysed with the Chi square test and the test of difference between two proportions. We calculated the standard mortality ratio (SMR), the mortality rates per 100 person-years (P-Y), and the death probability according to Grevile's formula, both in the population of Bologna and in our DA population. We observed 182 deaths (14.99%) among the 1214 monitored over a 8012 person-year period. From 1984 to 1997 the mortality rate in the DA population was 2.27 per 100 P-Y. In the 1986-1996 period the death probability among DA was 4-to 5-fold higher than in the general population for the same age groups. From 1977 to 1983 no disease was recorded among the first 200 enrolled subjects. From 1984 to 1977 mortality was 28.1% among the 426 HIV positive DA, 0.46% among the 433 HIV-negative DA, and 17.1% among the 355 DA with unknown HIV status. AIDS caused 106 deaths (58%), overdoses 40 (22%), car accident 15 (8%), hepatic cirrhosis 9 (5%), and other causes (miscellany) 11 (6%). In 1 (0.5%) case the cause of death was unknown. The mortality rate (MR) for AIDS was 12.8% for females, 7.4% for males (p<0.05); the MR for overdose was 3.8% for males, 0% for females; the MR for car accident was 1.2% for males, 1.3% for females; the MR for the miscellany of other causes was 1.3% for males and 0% for females. The SMR in the 20-49 age group in the 1984-1990 period was 34.7, while in the 1991-1996 period it was 143.4. PMID- 12728203 TI - [Survey on ciprofloxacin resistance: the epidemiological situation in Pordenone (Italy)] AB - An epidemiological survey on 2859 bacterial strains was performed in the first nine months of 1998, to verify the predominant susceptibility to ciprofloxacin of the main microrganisms isolated in this period. Our results showed that the use of ciprofloxacin would probably decrease in future because of increasing resistance with respect to previous period. An increase in antimicrobial resistance occurred in some gram positive (as Enterococci) and especially gram negative strains such as Escherichia coli, that are common and frequent bacterial causes of infections in hospital and in the community. The study demonstrated that there are also interesting and important variations in resistance patterns with respect to previous studies PMID- 12728191 TI - Excitotoxic and excitoprotective mechanisms: abundant targets for the prevention and treatment of neurodegenerative disorders. AB - Activation of glutamate receptors can trigger the death of neurons and some types of glial cells, particularly when the cells are coincidentally subjected to adverse conditions such as reduced levels of oxygen or glucose, increased levels of oxidative stress, exposure to toxins or other pathogenic agents, or a disease causing genetic mutation. Such excitotoxic cell death involves excessive calcium influx and release from internal organelles, oxyradical production, and engagement of programmed cell death (apoptosis) cascades. Apoptotic proteins such as p53, Bax, and Par-4 induce mitochondrial membrane permeability changes resulting in the release of cytochrome c and the activation of proteases, such as caspase-3. Events occurring at several subcellular sites, including the plasma membrane, endoplasmic reticulum, mitochondria and nucleus play important roles in excitotoxicity. Excitotoxic cascades are initiated in postsynaptic dendrites and may either cause local degeneration or plasticity of those synapses, or may propagate the signals to the cell body resulting in cell death. Cells possess an array of antiexcitotoxic mechanisms including neurotrophic signaling pathways, intrinsic stress-response pathways, and survival proteins such as protein chaperones, calcium-binding proteins, and inhibitor of apoptosis proteins. Considerable evidence supports roles for excitotoxicity in acute disorders such as epileptic seizures, stroke and traumatic brain and spinal cord injury, as well as in chronic age-related disorders such as Alzheimer's, Parkinson's, and Huntington's disease and amyotrophic lateral sclerosis. A better understanding of the excitotoxic process is not only leading to the development of novel therapeutic approaches for neurodegenerative disorders, but also to unexpected insight into mechanisms of synaptic plasticity. PMID- 12728205 TI - HIV disease and bacterial superinfections due to Xanthomonas spp.: a frequent association. AB - Sixty-four episodes of Xanthomonas spp. infection were observed in 2.400 patients hospitalised for HIV disease (~2.7%) over an 8-year period: sepsi-bacteremia in 52 cases, lower respiratory tract infection in 6 cases, urinary tract infection in three patients, pharyngitis in two cases, and lymph node resented the fourth most common non-mycobacterial bacterial pathogens responsible for bacteremia in our HIV-infected patients: 52 cases out of 878 diagnosed (5.9%). The progression of HIV-related immunodeficiency, the occurrence of leukopenia-neutropenia, hospitalisation, previous antibiotic and/or corticosteroid treatment, and instrumentation, seemed to act as risk factors for the occurrence of Xanthomonas spp. infection. In three patients suffering from severe immunodeficiency and concurrent AIDS-related disorders, Xanthomonas spp. complication contributed to death, while a relapsing disease occurred in two cases only. Because of the poor antimicrobial susceptibility of these pathogens (also confirmed in our series), Xanthomonas spp. infection associated with advanced HIV disease and concurrent risk factors, may represent a potentially severe complication. PMID- 12728204 TI - Conservative treatment of diabetic foot infections. AB - Foot infections commonly occur in diabetic patients, constituting a serious problem. 15% of diabetic have foot infections during their life-time. In previous years these life- threatening infections in 50-70% of patients resulted in limb amputation. Today, using an aggressive surgical approach, broad spectrum antimicrobial chemotherapy and intensive regulation of blood glucose level, the amputation rate has decreased to 5%. During the last 2 years, we evaluated 13 diabetic foot infections. 6 of the presented cases were grade 4 and 5, 3 of them grade 3 and 4 of them grade 2. All of the patients were managed by multidisciplinary approach consisting of plastic and reconstructive surgery, endocrinology and infectious diseases specialists. All of the patients were monitored with daily wound care, regulation of blood glucose levels and administration of broad spectrum antibiotics. Only one of them required amputation. For 12 of the cases after the infection was controlled, the defects were reconstructed by plastic surgical procedures. We retrospectively analysed them to review diabetic foot infections and factors affecting prognosis. PMID- 12728206 TI - Salmonella typhi disease in HIV-infected patients: case reports and literature review. AB - Two patients with AIDS and severe immunodeficiency developed typhoid fever. The diagnosis was confirmed by isolation of Salmonella typhi from blood cultures, while Widal's serum test isolates proved in vitro sensitive to all tested antimicrobial agents, and complete recovery was obtained with i.v. cotrimoxazole and piperacillin treatment. Unlike complications by non-tiphoid Salmonellae, S. typhi infection has been infrequently reported in the context of AIDS: to our knowledge, only five cases of typhoid fever have been described in HIV-infected living industrialized countries. Medical history and epidemiological information may play an important role in prompting the search for an S. typhi infection in patients coming from endemic regions, since most of the sign and symptoms of typhoid fever may mimic those of a sepsis. Or other AIDS-related complications PMID- 12728207 TI - Primary dorsal localization of invertebrate hydatidosis. AB - We describe patient with Echinococcus granulosus disease primary localized on the back. This case is uncommon because of the lack of risk exposure history and the sole muscular localization. The patient received several surgical operations on the back, without eradication of the infestation. At the end we discuss the clinical considerations that induced the choice of the usual medical therapy PMID- 12728208 TI - Strascino's compliant. Syphilis and the XVI Century Sienese theatre. AB - Niccolo Campani, known with the nickname "Strascino" (as shabby, beggarly), is among the most important poets in the XVI Century in Italian theatre. Famous for his sharp-witted humor and the perfect Tuscan rhyming, the Sienese poet left a work whose subject is syphilis, cooght by Campany from 1503 to 1511. In Strascino's Compliant, the poet describes in rhyme the course and recovery of the illness, giving a very personal testimony of being ill with the "la malaise francaise" and a broad overview of the sixteenth Century in Italy PMID- 12728209 TI - 4th Congress Bacterial Infections and Therapeutic Perspectives (IBAT) 1999 in Naples. AB - Not available PMID- 12728210 TI - Ethical conflicts of interest. PMID- 12728211 TI - Update on adequacy of protein and energy intake during pregnancy. PMID- 12728212 TI - Public policy: protecting what we've achieved. PMID- 12728213 TI - Qualitative vs quantitative methods: two opposites that make a perfect match. PMID- 12728214 TI - Selected lower-fat foods positively impact nutrient quality in diets of free living Americans. AB - OBJECTIVE: To compare nutrient profiles of Continuing Survey of Food Intake by Individuals (CSFII) respondents who reported use of regular and lower-fat versions of selected foods with those not reporting intake of these foods. SUBJECTS: American children (ages 2 to 19) and adults completing two days of recalls for the 1994 to 1996 CSFII. DESIGN: Dietary intakes were reviewed for selected foods for which regular (high) and modified-fat versions were available. Foods included added table fats (eg, salad dressings, margarine, creams), desserts, snacks, cheeses, and yogurts. Respondents were divided into non-users (none of these foods reported), mixed users (lower-fat and high-fat foods), and high-fat (only) users. STATISTICAL ANALYSES PERFORMED: Weighted data were used for descriptive analyses. Unweighted data were used for reported nutrient intake, nutrient density, fat intakes, and Food Guide Pyramid comparisons. A P value of < or =.01 was selected to determine whether group differences were statistically significant. Post hoc analyses were conducted to identify where differences were found. RESULTS: Respondents reporting inclusion of any selected lower-fat food had significantly lower fat intake (P<.01) than exclusive high-fat users (male respondents: 31.5% vs 35.1%; female respondents: 30.3% vs 34.4%; children: 30.6% vs 33.4%). In general, mixed users had higher micronutrient intakes than high-fat users. Specifically, mixed users reported higher intakes of most B vitamins; vitamins A, C, and folate; the minerals calcium, phosphorus, magnesium, iron, and zinc; and fiber. These differences were associated with higher intakes of milk, fruits, and grains. APPLICATIONS/CONCLUSIONS: This research supports the "all foods can fit" paradigm and validates recommending lower-fat foods (including snacks, desserts, cheeses, yogurt, and added table fats) to American consumers to reduce total fat to moderate levels while ensuring adequacy for most micronutrients. PMID- 12728215 TI - Fat-free foods supplemented with soy stanol-lecithin powder reduce cholesterol absorption and LDL cholesterol. AB - OBJECTIVE: The objective of this work was to show that fat-free, lecithin formulated soy stanols lower cholesterol absorption and serum LDL cholesterol. DESIGN: Reduction in cholesterol absorption was measured in paired single-meal tests with or without formulated soy stanols (acute test), and changes in serum lipids were investigated in a 10-week, randomized, double-blind parallel trial in which formulated stanols or lecithin vehicle were given three times daily for the last 4 weeks (chronic test). SUBJECTS/SETTING: Forty-five normal or mildly hypercholesterolemic subjects were recruited for both studies. The 21 subjects (16 female, 5 male; mean age 32.5 years) in the absorption studies had the following mean lipid values: LDL cholesterol, 2.79 mmol/L and total cholesterol, 4.73 mmol/L. For the lipid reduction, 24 subjects (16 female, 8 male; mean age 50.6 years) were enrolled with mean LDL cholesterol and total cholesterol of 3.72 mmol/L and 5.66 mmol/L, respectively. INTERVENTION: Reduction in cholesterol absorption was measured using a lemonade beverage or egg whites that contained 625 mg stanols. Throughout the chronic study, subjects followed the American Heart Association Step I diet. During the 4-week treatment phase, subjects consumed daily a lemonade-flavored beverage containing either placebo or formulated soy stanols (1.9 g). MAIN OUTCOME MEASURES: Inhibition of cholesterol absorption was determined from the difference in plasma deuterated cholesterol enrichment after a test meal containing stanol-lecithin and one with lecithin vehicle only. In the chronic study, the primary endpoints were changes in LDL and total cholesterol. STATISTICAL ANALYSES PERFORMED: Paired or unpaired t tests were used to determine statistical significance. RESULTS: Stanol-lecithin reduced cholesterol absorption by 32.1% (P=.0045, n=10) and by 38.2% (P=.0022, n=11) when delivered in a lemonade-flavored beverage and in egg whites, respectively. Reduction in cholesterol absorption was strongly related to the initial level of absorbed cholesterol tracer in serum (r(s)=-0.739). Stanol-lecithin given in a beverage reduced total serum cholesterol by 10.1% (P=.0019, n=24) and LDL cholesterol by 14.3% (P=.0016, n=24). APPLICATIONS/CONCLUSIONS: Powdered soy stanol-lecithin lowers cholesterol absorption and LDL cholesterol when consumed in fat-free foods. PMID- 12728216 TI - Kefir improves lactose digestion and tolerance in adults with lactose maldigestion. AB - OBJECTIVE: Kefir is a fermented milk beverage that contains different cultures than yogurt. The objective of this study was to determine whether kefir improves lactose digestion and tolerance in adults with lactose maldigestion. DESIGN: Randomized block design. SUBJECTS: Fifteen healthy, free-living adults with lactose maldigestion. MAIN OUTCOME MEASURES: Breath hydrogen excretion and lactose intolerance symptoms were monitored hourly for 8 hours after each test meal. INTERVENTION: Subjects were fed test meals consisting of 20 g lactose portions of milk (2% reduced fat), plain and raspberry flavored kefir, and plain and raspberry flavored yogurt, each following an overnight (12 hour) fast. STATISTICAL ANALYSIS: Mixed model ANOVA was performed on raw or transformed data, followed by Tukey HSD post hoc tests (when appropriate). Significance was defined as P<.05. RESULTS: The breath hydrogen area under the curve (AUC) for milk (224+/ 39 ppm x h) was significantly greater than for the plain yogurt (76+/-14 ppm x h, P<.001), the plain kefir (87+/-37 ppm x h, P<.001), and the flavored yogurt (76+/ 14 ppm x h, P=.005). The flavored kefir had an intermediate response (156+/-26 ppm x h). The yogurts and kefirs all similarly reduced the perceived severity of flatulence by 54% to 71% relative to milk. Abdominal pain and diarrhea symptoms were negligible among the five treatments. APPLICATIONS/CONCLUSION: Because kefir improved lactose digestion and tolerance in this study, its use may be another potential strategy for overcoming lactose intolerance. Further studies of other types of kefir for improving lactose digestion are warranted. PMID- 12728217 TI - A new handheld device for measuring resting metabolic rate and oxygen consumption. AB - OBJECTIVE: The purpose of this study was to test a new handheld device, the BodyGem (HealtheTech Inc., Golden, CO), that measures resting metabolic rate (RMR). SUBJECTS/DESIGN: Sixty-three adults (43 women, 20 men) (mean+/-SD, age 41.3+/-11.2 years and body mass index, 26.5+/-6.6 kg/m(2)) were tested during two separate sessions within a 2-week period, and, in each session, two BodyGem and two Douglas bag RMR measurements were made in a random and counterbalanced order. MAIN OUTCOME MEASURES: Resting oxygen consumption and energy expenditure. STATISTICAL ANALYSES PERFORMED: A 2 (methods) x 4 (time points) repeated measures ANOVA, Pearson product-moment coefficients, Bland-Altman plots. RESULTS: Within session reliability for measurement of O(2), consumption was high on both days for the BodyGem (r=0.97). During the 4 single tests, BodyGem and Douglas bag O(2) consumption values were significantly correlated (r=0.81-0.87), with SEEs ranging from 22 to 28 mL. min(-1). Mean O(2) consumption and RMR values for all 4 tests were 241+/-46 and 240+/-45 mL x min(-1) (r=0.91, SEE 18.7 mL x min(-1)) and 1,657+/-324 and 1,650+/-307 kcals x day(-1) (r=0.91, SEE 134 kcals x day(-1)) for the BodyGem and Douglas bag methods, respectively. APPLICATIONS/CONCLUSIONS: These data indicate that the BodyGem is an accurate and reliable device for measuring oxygen consumption and calculating RMR during repeated tests within a day, single tests on separate days, or when measurements are averaged. PMID- 12728218 TI - Is there a role for diet in the treatment of hemachromatosis? PMID- 12728220 TI - Development of a master's in public health nutrition degree program using distance education. AB - The Professional Practice Program in Nutrition (PPPN) uses distance education to offer a master's degree in public health (MPH) nutrition designed for practitioners who desire to maintain their employment and develop new skills. Public health nutrition leadership faces challenges in recruiting a large enough workforce to (a) carry out the core functions of assessment, policy, and assurance; (b) update current nutritionists in new skill areas to face the demands of dynamic health care and public health climates; and (c) conduct monitoring and surveillance of Healthy People 2010 objectives. In 1996, the University of North Carolina at Chapel Hill designed and implemented this program after a market analysis to identify advanced educational need in a nine state area. PPPN was initiated as a pilot program and enrolled two cohorts of students from 1996 through 1998. This project identified four key steps: (a) conduct a detailed market analysis; (b) establish an infrastructure to deliver the program; (c) tailor the curriculum using the technology; and (d) identify, accommodate, and develop student capabilities. The findings indicate that distance education strategies are appropriate to carry out a full MPH curriculum in nutrition, but sufficient enrollment is necessary to cover the added curriculum expenditures. PMID- 12728219 TI - Comparisons of vegetarian and beef-containing diets on hematological indexes and iron stores during a period of resistive training in older men. AB - OBJECTIVE: To test the hypothesis that older men who consumed a vegetarian (lacto ovo) diet would develop a lower iron status compared with older men who consumed a beef-containing diet during a period of resistive training (RT). DESIGN: Experimental, repeated measures study. SUBJECTS: Twenty-one healthy men aged 59 to 78 years, with a BMI range of 24 to 33 kg/m(2), completed the study. INTERVENTION: All men consumed a vegetarian diet for 2 weeks (baseline). After this, the men were randomly assigned to one of two dietary groups. Eleven men consumed a beef-containing diet, and 10 men continued to consume a vegetarian diet for 12 weeks. During this time all subjects participated in RT three days per week, designated as RT1 to RT12. MAIN OUTCOME MEASURES: Serum ferritin and serum iron concentrations, transferrin saturation, transferrin receptor, total iron binding capacity, and selected hematological variables, as well as selected nutrient intakes and estimated iron bioavailability from three-day diet records, were determined at baseline, RT5, and RT12. STATISTICAL ANALYSES: A general linear model repeated-measures ANOVA was used to examine the effects of group, time, and group x time interactions for iron status and dietary data. RESULTS: Total iron intake was not different between the two groups; however, the beef group had a three to four times greater intake of bioavailable iron (P<.01) than the vegetarian group. Serum iron, total iron binding capacity, transferrin saturation, and transferrin receptor were not significantly different between the beef and vegetarian groups, or changed over time with RT. Serum ferritin decreased over time in both the beef and vegetarian groups during RT (P<.01). Re introduction of beef into the diets of the beef group increased hemoglobin concentration and hematocrit compared with the vegetarian group during the 12 weeks of RT (group x time, P<.05). These changes were within clinically normal limits. APPLICATIONS/CONCLUSIONS: Older men who consume a beef-containing, higher bioavailable-iron diet, compared with a vegetarian, lower-bioavailable-iron diet, have an increased hematological profile during a 12-week period of RT. Older men who consume either a beef-containing or a vegetarian diet maintain a hematological profile within clinically normal limits during 12 weeks of RT. PMID- 12728221 TI - Integrating issues of sustainably produced foods into nutrition practice: a survey of Minnesota Dietetic Association members. AB - A survey was mailed to a representative, randomly selected sample of Minnesota Dietetic Association members (n=300) to evaluate attitudes and intentions regarding the integration of issues pertaining to sustainably produced foods into professional practice. The United States Department of Agriculture describes sustainable agriculture as including goals of farm profitability, environmental stewardship, and improved rural quality of life. Data analysis included descriptive statistics, independent t tests, one-way ANOVA, correlations, and multiple regression analyses (P< or =.05). Forty-three percent had heard of sustainable agriculture, 13% had integrated issues of sustainably produced foods into their practice, and 20% stated that it was likely that they would incorporate these issues within the next six months. Perceived behavioral control, subjective norm, and rated importance of knowing about agricultural practices independently predicted intention. Dietitians surveyed were interested in learning more, but most were not integrating these issues into practice because of perceived barriers, including a lack of knowledge about this topic. PMID- 12728222 TI - Pregnant adolescents in Guam consume diets low in calcium and other micronutrients. AB - The purpose of this study was to assess diets of pregnant adolescents in Guam, including difference in age and trimester. The 434 subjects, aged 14 to 20 years, were primarily Chamorro and Filipino. Analysis of variance was used to determine differences in nutrient intake, and Tukey post hoc testing was used. Frequency distributions were calculated for intake at various dietary reference intake levels. Mean energy intake was 2,487+/-1,057 kcal. The nutrients that are inadequate in the diets of the adolescents are calcium, folate, vitamin E, magnesium, and iron. Calcium intake declined with increasing age (P<.0001). Notable differences (P<.01) among trimester existed only for iron, which was lower in the second than third trimester. Nutrient intake is similar to that in pregnant adolescents in the United States, with particularly low calcium levels. Culturally specific materials emphasizing nutrition during pregnancy and chronic disease prevention are warranted. PMID- 12728223 TI - Position of the American Dietetic Association: Oral health and nutrition. AB - It is the position of the American Dietetic Association (ADA) that nutrition is an integral component of oral health. The ADA supports the integration of oral health with nutrition services, education, and research. Collaboration between dietetics and dental professionals is recommended for oral health promotion and disease prevention and intervention. Scientific and epidemiological data suggest a lifelong synergy between nutrition and the integrity of the oral cavity in health and disease. Oral health is an integral part of systemic and nutritional health. Two primary oral infectious diseases are directly influenced by diet and nutrition. Dental caries or tooth decay is modulated by numerous factors, including diet composition and frequency. Periodontal or gum disease is associated with malnutrition. Chronic diseases such as diabetes and cardiovascular disease that are modulated by diet and nutrition intervention have oral sequelae. As we advance in our discoveries of the links between oral and nutrition health, practitioners of both disciplines must learn to provide screening, baseline education, and referral to each other as part of comprehensive client/patient care. The future of dietetics practice requires dietetics professionals to provide medical nutrition therapy (MNT) that incorporates a person's total health needs, including oral health. Inclusion of both didactic and clinical practice concepts that illustrate the role of nutrition in oral health is essential in both dental and dietetic education programs. Collaborative endeavors between dietetics and dentistry in research, education, and delineation of health provider practice roles are needed to ensure comprehensive health care to persons with oral infectious disease and/or oral manifestations of systemic diseases. PMID- 12728224 TI - Blending practice and research: Practice-based research networks an opportunity for dietetics professionals. PMID- 12728226 TI - Hard choices. PMID- 12728227 TI - From bench to business. PMID- 12728225 TI - Ethics opinion: Dietetics professionals are ethically obligated to maintain personal competence in practice. PMID- 12728229 TI - Irradiated materials: Defects break symmetry rules. PMID- 12728230 TI - Ferromagnetic semiconductors: A giant appears in spintronics. PMID- 12728231 TI - Fuel cells: The amazing perovskite anode. PMID- 12728232 TI - Nanoscale photonics: Nanoshells: gifts in a gold wrapper. PMID- 12728233 TI - Actuator materials: Towards smart artificial muscle. PMID- 12728235 TI - Self-cleaning surfaces--virtual realities. AB - In the 19th century, Oscar Wilde stated "We live, I regret to say, in an age of surfaces". Today, we do so even more, and we do not regret it: key advances in the understanding and fabrication of surfaces with controlled wetting properties are about to make the dream of a contamination-free (or 'no-clean') surface come true. Two routes to self-cleaning are emerging, which work by the removal of dirt by either film or droplet flow. Although a detailed understanding of the mechanisms underlying the behaviour of liquids on such surfaces is still a basic research topic, the first commercial products in the household-commodity sector and for applications in biotechnology are coming within reach of the marketplace. This progress report describes the current status of understanding of the underlying mechanisms, the concepts for making such surfaces, and some of their first applications. PMID- 12728238 TI - The European Research Council: where are we now? PMID- 12728239 TI - DNA electronics. PMID- 12728240 TI - The way we write. PMID- 12728241 TI - Looking for the trigger. The Environmental Genome Project to uncover the interactions of genes and the environment in disease. PMID- 12728242 TI - Science and sustainability. More research is paramount for finding sustainable solutions for agriculture in developing countries. But politics has an equally important role. PMID- 12728245 TI - Outer mitochondrial membrane permeabilization: an open-and-shut case? PMID- 12728244 TI - The AVIT protein family. Secreted cysteine-rich vertebrate proteins with diverse functions. AB - Homologues of a protein originally isolated from snake venom and frog skin secretions are present in many vertebrate species. They contain 80-90 amino acids, 10 of which are cysteines with identical spacing. Various names have been given to these proteins, such as mamba intestinal protein 1 (MIT1), Bv8 (Bombina variegata molecular mass approximately 8 kDa), prokineticins and endocrine-gland vascular endothelial growth factor (EG-VEGF). Their amino-terminal sequences are identical, and so we propose that the sequence of their first four residues, AVIT, is used as a name for this family. From a comparison of the sequences, two types of AVIT proteins can be discerned. These proteins seem to be distributed widely in mammalian tissues and are known to bind to G-protein-coupled receptors. Members of this family have been shown to stimulate contraction of the guinea pig ileum, to cause hyperalgesia after injection into rats and to be active as specific growth factors. Moreover, the messenger RNA level of one of these AVIT proteins changes rhythmically in the region of the brain known as the suprachiasmatic nucleus. This shows that members of this new family of small proteins are involved in diverse biological processes. PMID- 12728246 TI - Mitochondrial functions during cell death, a complex (I-V) dilemma. PMID- 12728247 TI - Realizing the promise of apoptosis-based therapies: separating the living from the clinically undead. PMID- 12728248 TI - DeltaFosB, but not FosB, induces delayed apoptosis independent of cell proliferation in the Rat1a embryo cell line. AB - The fates of Rat1a cells expressing FosB and DeltaFosB as fusion proteins (ER FosB, ER-DeltaFosB) with the ligand binding domain of human estrogen receptor were examined. The binding of estrogen to the fusion proteins resulted in their nuclear translocation and triggered cell proliferation, and thereafter delayed cell death was observed only in cells expressing ER-DeltaFosB. The proliferation of Rat1a cells, but not cell death triggered by ER-DeltaFosB, was completely abolished by butyrolactone I, an inhibitor of cycline-dependent kinases, and was partly suppressed by antisense oligonucleotides against galectin-1, whose expression is induced after estrogen administration. The cell death was accompanied by the activation of caspase-3 and -9, the fragmentation of the nuclear genome and cytochrome c release from the mitochondria, and was suppressed by zDEVD-fmk and zLEHD-fmk but not zIETD-fmk. The cell death was not suppressed by exogenous His-PTD-Bcl-x(L) at all, suggesting involvement of a Bcl-x(L) resistant pathway for cytochrome c release. PMID- 12728249 TI - Inhibition of hypoxia/reoxygenation-induced oxidative stress in HGF-stimulated antiapoptotic signaling: role of PI3-K and Akt kinase upon rac1. AB - Rac1-regulated reactive oxygen species (ROS) production has been implicated in apoptosis. In contrast, pleiotropic protein kinase Akt protects against apoptosis. However, the pro- and antiapoptotic mechanisms of rac1 and Akt, respectively, and the intersection between these mechanisms are incompletely understood. In a model of oxidative stress and apoptosis induced by hypoxia/reoxygenation (H/R) in primary hepatocytes, activation of the PI3-K Akt axis by the prosurvival hepatocyte growth factor (HGF) inhibited H/R-stimulated rac1 activation and intracellular ROS production, and suppressed apoptosis. Suppression of PI3-K or Akt activity abrogated the inhibitory effect of HGF on rac1 activity and rac1-regulated oxidative stress. Furthermore, constitutive activation of Akt or PI3-K in the absence of HGF was sufficient to phosphorylate rac1, inhibit rac1 activation, and suppress rac1-regulated ROS production. These findings demonstrate that growth factor-stimulated activation of PI3-K-Akt is necessary and sufficient to suppress intracellular oxidative stress and apoptosis by inhibiting activation of pro-apoptotic, prooxidative rac1 GTPase. PMID- 12728250 TI - TGF-beta1 suppresses apoptosis via differential regulation of MAP kinases and ceramide production. AB - Serum deprivation induces apoptosis in NIH3T3 cells, which is associated with increased intracellular ceramide generation and with the activation of p38 mitogen-activated protein (MAP) kinase. Treatment of cells with transforming growth factor-beta1 (TGF-beta1) activated the extracellular signal regulated kinases 1 and 2 (ERK1/ERK2), inhibited the serum deprivation-induced p38 activation and the increase in intracellular ceramide formation, leading to the stimulation of cell proliferation and the suppression of apoptosis. Inhibition of p38 MAP kinase by SB203580 significantly reduced the serum-deprivation-induced apoptosis. Overexpression of p38 increased the cell apoptosis and reduced the antiapoptotic effect of TGF-beta1. Inhibition of ERK1/ERK2 by PD98059 completely inhibited the TGF-beta1-stimulated proliferation and partially inhibited the antiapoptotic effects of TGF-beta1. Neither SB203580 nor PD98059 has obvious effect on TGF-beta1-mediated inhibition of the increased ceramide generation. Serum-deprivation-induced apoptosis in NIH3T3 cells can also be blocked by broad spectrum caspase inhibitor. TGF-beta1 treatment has an inhibitory effect on caspase activities. Our results indicate that ceramide, p38, and ERK1/ERK2 play critical but differential roles in cell proliferation and stress-induced apoptosis. TGF-beta1 suppresses the serum-deprivation-induced apoptosis via its distinct effects on complex signaling events involving the activation of ERK1/ERK2 and the inhibition of p38 activation and increased ceramide generation. PMID- 12728251 TI - Role of DDC-4/sFRP-4, a secreted frizzled-related protein, at the onset of apoptosis in mammary involution. AB - Using differential display, we isolated DDC-4, a secreted frizzled-related protein (sFRP), which is induced in the physiological apoptosis of hormonally regulated, reproductive tissues such as mammary gland, prostate, corpus luteum and uterus. The role of this gene in apoptosis was studied in animals overexpressing ectopic DDC-4/sFRP-4. Transgenic mice bearing the DDC-4/sFRP-4 cDNA under the control of the MMTV-LTR promoter showed lactational insufficiency and many apoptotic cells in the alveoli between day 19 of pregnancy and day 4 of lactation as demonstrated by TUNEL reaction and the presence of activated caspase 3. We performed a PKB/Akt kinase assay and studied several of its substrates using phosphorylation-specific antibodies to show reduced phosphorylation in PKB/Akt itself, as well as in glycogen synthetase kinase-3beta (GSK-3beta), BAD, and Forkhead. Taken together, our results show a role for DDC-4/sFRP-4 in abrogating an epithelial cell survival pathway at the onset of mammary gland involution. PMID- 12728252 TI - Seizure-like activity leads to the release of BAD from 14-3-3 protein and cell death in hippocampal neurons in vitro. AB - Seizure-induced neuronal death may involve engagement of the BCL-2 family of apoptosis-regulating proteins. In the present study we examined the activation of proapoptotic BAD in cultured hippocampal neurons following seizures induced by removal of chronic glutamatergic transmission blockade. Kynurenic acid withdrawal elicited an increase in seizure-like electrical activity, which was inhibited by blockers of AMPA (CNQX) and NMDA (MK801 and AP5) receptor function. However, only NMDA receptor antagonists inhibited calcium entry as assessed by fura-2, and cell death of hippocampal neurons. Seizures increased proteolysis of caspase-3 and terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) of cells. Seizure-like activity induced dephosphorylation of BAD and the disruption of its constitutive interaction with 14-3-3 proteins. In turn, BAD dimerized with antiapoptotic BCL-Xl after seizures. However, the absence of neuroprotective effects of pathway intervention suggests that BAD may perform a reinforcement rather than instigator role in cell death following seizures in vitro. PMID- 12728253 TI - Apoptosis induced by Na+/H+ antiport inhibition activates the LEI/L-DNase II pathway. AB - L-DNase II is derived from its precursor leucocyte elastase inhibitor (LEI) by post-translational modification. In vitro, the conversion of LEI into L-DNase II can be induced by incubation of LEI at an acidic pH. In this study, we proposed to analyze the effects of intracellular acidification on this transformation. Amiloride derivatives, like hexamethylene amiloride (HMA), are known to provoke a decrease of cytosolic pH by inhibiting the Na(+)/H(+) antiport. In BHK cells, treatment with HMA-induced apoptosis accompanied by an increase in L-DNase II immunoreactivity and L-DNase II enzymatic activity. Overexpression of L-DNase II precursor led to a significant increase of apoptosis in these cells supporting the involvement of L-DNase II in HMA induced apoptosis. As previously shown in other cells, etoposide-induced apoptosis did not activate L-DNase. On the contrary, LEI overexpression significantly increased cell survival in etoposide induced apoptosis. Together these results suggest differential roles of LEI and L DNase II in response to different types of apoptotic inducers. PMID- 12728255 TI - Caspase-mediated cleavage of the U snRNP-associated Sm-F protein during apoptosis. AB - Recent studies have implicated the dying cell as a potential reservoir of modified autoantigens that might initiate and drive systemic autoimmunity in susceptible hosts. The spliceosomal Sm proteins are recognized by the so-called anti-Sm autoantibodies, an antibody population found exclusively in patients suffering from systemic lupus erythematosus. We have studied the effects of apoptosis on the Sm proteins and demonstrate that one of the Sm proteins, the Sm F protein, is proteolytically cleaved in apoptotic cells. Cleavage of the Sm-F protein generates a 9-kDa apoptotic fragment, which remains associated with the U snRNP complexes in apoptotic cells. Sm-F cleavage is dependent on caspase activation and the cleavage site has been located near the C-terminus, EEED(81) downward arrow G. Use of different caspase inhibitors suggests that besides caspase-8 other caspases are implicated in Sm-F cleavage. A C-terminally truncated mutant of the Sm-F protein, representing the modified form of the protein, is capable of forming an Sm E-F-G complex in vitro that is recognized by many anti-Sm patient sera. PMID- 12728254 TI - An MMP-2/MMP-9 inhibitor, 5a, enhances apoptosis induced by ligands of the TNF receptor superfamily in cancer cells. AB - Several studies have shown that matrix metalloproteases (MMPs) promote tumor growth, invasion, and metastasis. Consequently, MMP inhibitors have been developed as a new class of anticancer drugs, many of which are in clinical trials. The exact mechanism of the antineoplastic activity of MMP antagonists is unknown. To investigate the mechanism, we hypothesized that MMP inhibitors enhance the actions of apoptosis-inducing agents. To test this hypothesis, we treated breast, melanoma, leukemia, osteosarcoma, and normal breast epithelial cells with (2R)-2-[(4-biphenylsulfonyl)amino]-3-phenylproprionic acid (compound 5a), an organic inhibitor of MMP-2/MMP-9, alone or in combination with TNFalpha or other apoptotic agents. FACS analysis showed that 5a interacted synergistically with ligands of the TNF receptor superfamily, including TNFalpha and TNF receptor-like apoptosis-inducing ligand (TRAIL), and with a Fas-cross linking antibody (CH11), UV, paclitaxel, thapsigargin, and staurosporin, to induce apoptosis in a cell-type-specific manner. Other MMP inhibitors did not synergize with TNFalpha. Compound 5a did not act directly on the mitochondrion or via changes in protein synthesis. Instead, the mechanism requires ligand-receptor interaction and caspase 8 activation. Investigation of the effect of 5a on tumor growth in vivo revealed that continuous treatment of subcutaneous melanoma with a combination of 5a plus TRAIL reduced tumor growth and angiogenesis in nude mice. Our data demonstrate that 5a possesses a novel proapoptotic function, thus providing an alternative mechanism for its antineoplastic action. These observations have important implications for combination cancer therapy. PMID- 12728257 TI - Bcr-Abl-mediated resistance to apoptosis is independent of constant tyrosine kinase activity. AB - Bcr-Abl is one of the most potent antiapoptotic molecules and is the tyrosine kinase implicated in Philadelphia (Ph) chromosome-positive leukemia. It is still obscure how Bcr-Abl provides the leukemic cell a strong resistance to chemotherapeutic drugs. A rational drug development produced a specific inhibitor of the catalytic activity of Bcr-Abl called STI571. This drug was shown to eliminate Bcr-Abl-positive cells both in vitro and in vivo, although resistant cells may appear in culture and relapse occurs in some patients. In the study described here, Bcr-Abl-positive cells treated with tyrosine-kinase inhibitors such as herbimycin A, genistein or STI571 lost their phosphotyrosine-containing proteins, but were still extremely resistant to apoptosis. Therefore, in the absence of tyrosine-kinase activity, Bcr-Abl-positive cells continue to signal biochemically to prevent apoptosis induced by chemotherapeutic drugs. We propose that secondary antiapoptotic signals are entirely responsible for the resistance of Bcr-Abl-positive cells. Precise determination of such signals and rational drug development against them should improve the means to combat Ph chromosome positive leukemia. PMID- 12728256 TI - NRADD, a novel membrane protein with a death domain involved in mediating apoptosis in response to ER stress. AB - NRADD (neurotrophin receptor alike death domain protein) is a novel protein with transmembrane and cytoplasmic regions highly homologous to death receptors, particularly p75(NTR). However, the short N-terminal domain is unique. Expression of NRADD induced apoptosis in a number of cell lines. The apoptotic mechanism involved the activation of caspase-8 and execution of apoptosis without requiring mitochondrial components. The activation of this death receptor-like mechanism required the N-terminal domain, which is N-glycosylated and needed for subcellular targeting. Deletion of the N-terminal domain produced a dominant negative form of NRADD that protected neurons and Schwann cells from a variety of endoplasmic reticulum (ER) stressors. NRADD may therefore be a necessary component for generating an ER-induced proapoptotic signal. PMID- 12728259 TI - Activation-associated mitochondrial hyperpolarization hijacks T cells toward an apoptosis-sensitized phenotype. PMID- 12728258 TI - c-Myc induces cytochrome c release in Rat1 fibroblasts by increasing outer mitochondrial membrane permeability in a Bid-dependent manner. AB - Ectopic expression of c-myc sensitises cells to a wide range of apoptotic stimuli by inducing the release of cytochrome c from the mitochondrial intermembrane space into the cytosol. To elucidate the molecular mechanisms of mitochondrial permeabilisation in response to c-Myc activation, we carried out a biochemical fractionation analysis of Rat1 fibroblasts expressing an inducible c-Myc protein. We find that cytoplasmic extracts from cells in which c-Myc has been activated contain a soluble factor capable of inducing cytochrome c release from isolated mouse liver mitochondria. This factor is present only under growth factor deprivation conditions and its activity is inhibited by addition of Bcl-X(L). The c-Myc-induced factor copurifies with full-length Bid, a "BH3-only" proapoptotic member of the Bcl-2 family, and antibodies raised against the BH3 domain of Bid inhibit c-Myc-induced cytochrome c releasing activity. These results are consistent with a model in which the activation of c-Myc regulates factors capable of enhancing the mitochondrial membrane destabilisation function of "BH3 only" proteins. PMID- 12728260 TI - Increased DeltaN-p73 expression in tumors by upregulation of the E2F1-regulated, TA-promoter-derived DeltaN'-p73 transcript. PMID- 12728261 TI - Involvement of interferon regulatory factor-1 in monocyte CD95 expression and CD95-mediated apoptosis. PMID- 12728262 TI - Sleuthing molecular targets for neurological diseases at the neuromuscular junction. PMID- 12728263 TI - Viral vectors for gene delivery to the nervous system. PMID- 12728264 TI - Targeting programmed cell death in neurodegenerative diseases. PMID- 12728265 TI - Pathogenesis of bacterial meningitis: from bacteraemia to neuronal injury. PMID- 12728266 TI - Neurobiology of migraine. PMID- 12728267 TI - Mechanisms, challenges and opportunities in stroke. PMID- 12728268 TI - Brain-machine interfaces to restore motor function and probe neural circuits. PMID- 12728269 TI - Nuclear lipid signalling. AB - During the past twenty years, evidence has accumulated for the presence of phospholipids within the nuclei of eukaryotic cells. These phospholipids are distinct from those that are obviously present in the nuclear envelope. The best characterized of the intranuclear lipids are the inositol lipids that form the components of a phosphoinositide-phospholipase C cycle. However, exactly as has been discovered in the cytoplasm, this is just part of a complex picture that involves many other lipids and functions. PMID- 12728270 TI - Who's on first in the cellular response to DNA damage? AB - Cellular DNA-repair pathways involve proteins that have roles in other DNA metabolic processes, as well as those that are dedicated to damage removal. Several proteins, which have diverse functions and are not known to have roles in DNA repair, also associate with damaged DNA. These newly discovered interactions could either facilitate or hinder the recognition of DNA damage, and so they could have important effects on DNA repair and genetic integrity. The outcome for the cell, and ultimately for the organism, might depend on which proteins arrive first at sites of DNA damage. PMID- 12728271 TI - Ras proteins: different signals from different locations. AB - Ras signalling has classically been thought to occur exclusively at the inner surface of a relatively uniform plasma membrane. Recent studies have shown that Ras proteins interact dynamically with specific microdomains of the plasma membrane as well as with other internal cell membranes. These different membrane microenvironments modulate Ras signal output and highlight the complex interplay between Ras location and function. PMID- 12728272 TI - Phagocyte sabotage: disruption of macrophage signalling by bacterial pathogens. AB - Macrophages function at the front line of immune defences against incoming pathogens. But the ability of macrophages to internalize bacteria, migrate, recruit other immune cells to the site of infection and influence the nature of the immune response can provide unintended benefits for bacterial pathogens that are able to subvert or co-opt these normally effective defences. This review highlights recent advances in our understanding of the many interference strategies that are used by bacterial pathogens to undermine macrophage signalling. PMID- 12728273 TI - Sphingosine-1-phosphate: an enigmatic signalling lipid. AB - The evolutionarily conserved actions of the sphingolipid metabolite, sphingosine 1-phosphate (S1P), in yeast, plants and mammals have shown that it has important functions. In higher eukaryotes, S1P is the ligand for a family of five G-protein coupled receptors. These S1P receptors are differentially expressed, coupled to various G proteins, and regulate angiogenesis, vascular maturation, cardiac development and immunity, and are important for directed cell movement. PMID- 12728275 TI - Lipids on the frontier: a century of cell-membrane bilayers. AB - Our present picture of cell membranes as lipid bilayers is the legacy of a century's study that concentrated on the lipids and proteins of cell-surface membranes. Recent work is changing the picture and is turning the snapshot into a video. PMID- 12728274 TI - Coat proteins: shaping membrane transport. AB - Coat proteins allow the selective transfer of macromolecules from one membrane enclosed compartment to another by concentrating macromolecules into specialized membrane patches and then deforming these patches into small coated vesicles. Recent findings indicate that coat proteins might also participate in the differentiation of membrane domains within organelles and large transport carriers, as well as in the association of the carriers with the cytosketelon and with acceptor organelles. PMID- 12728276 TI - Integrating biological databases. AB - Recent years have seen an explosion in the amount of available biological data. More and more genomes are being sequenced and annotated, and protein and gene interaction data are accumulating. Biological databases have been invaluable for managing these data and for making them accessible. Depending on the data that they contain, the databases fulfil different functions. But, although they are architecturally similar, so far their integration has proved problematic. PMID- 12728277 TI - Progress and problems with the use of viral vectors for gene therapy. AB - Gene therapy has a history of controversy. Encouraging results are starting to emerge from the clinic, but questions are still being asked about the safety of this new molecular medicine. With the development of a leukaemia-like syndrome in two of the small number of patients that have been cured of a disease by gene therapy, it is timely to contemplate how far this technology has come, and how far it still has to go. PMID- 12728278 TI - What good is genomic imprinting: the function of parent-specific gene expression. AB - Parent-specific gene expression (genomic imprinting) is an evolutionary puzzle because it forgoes an important advantage of diploidy--protection against the effects of deleterious recessive mutations. Three hypotheses claim to have found a countervailing selective advantage of parent-specific expression. Imprinting is proposed to have evolved because it enhances evolvability in a changing environment, protects females against the ravages of invasive trophoblast, or because natural selection acts differently on genes of maternal and paternal origin in interactions among kin. The last hypothesis has received the most extensive theoretical development and seems the best supported by the properties of known imprinted genes. However, the hypothesis is yet to provide a compelling explanation for many examples of imprinting. PMID- 12728279 TI - Unique features of the plant life cycle and their consequences. AB - Continuous development, the absence of a germline, flexible and reversible cellular differentiation, and the existence of haploid and diploid generations- both of which express genes--are characteristics that distinguish plants from animals. Because these differences alter the impact of mutations, animals and plants experience varied selection pressures. Despite different life-cycles, both flowering plants and multicellular animals have evolved complex sensing mechanisms that act after fertilization as 'quality checks' on reproduction, and that detect chromosome dosage and the parent of origin for specific genes. Although flowering plant embryos escape such surveillance in vitro, embryo success in the seed often depends on a healthy endosperm--a nutritive tissue that is produced by a second fertilization event in which maternal and paternal gene contributions can be monitored immediately after fertilization and throughout development. PMID- 12728281 TI - Mitochondrial gene history and mRNA localization: is there a correlation? AB - Phylogenetic studies of the yeast mitochondrial proteome have shown a complex evolutionary scenario, in which proteins of bacterial origin form complexes with proteins of eukaryotic origin. Exciting new results from whole-genome microarray studies of subcellular mRNA localizations have shown that mRNAs that are of putative bacterial origin are mainly translated on polysomes that are associated with the mitochondrion, whereas those of eukaryotic origin are generally translated on free cytosolic polysomes. Understanding these newly discovered relationships promises insights into old questions about organelle origins and mRNA localization in the eukaryotic cell. PMID- 12728280 TI - Caenorhabditis elegans: an emerging genetic model for the study of innate immunity. AB - Invaluable insights into how animals, humans included, defend themselves against infection have been provided by more than a decade of genetic studies that have used fruitflies. In the past few years, attention has also turned to another simple animal model, the nematode worm Caenorhabditis elegans. What exactly have we learned from the work in Drosophila? And will research with C. elegans teach us anything new about our response to pathogen attack? PMID- 12728282 TI - Cutaneous leishmaniasis of the Old World. AB - Cutaneous Leishmaniasis is a vector-borne protozoal infection of the skin. Several species of Leishmania cause this disease in the Old World. It is manifested as chronic nodular to ulcerative lesions of the skin, which last for many months and may be disfiguring. They eventually heal leaving a scar. Local care of the lesion and treatment of secondary bacterial infection are essential for healing. Antileishmania therapy is indicated in immunocompromised hosts, patients with progressive, multiple, or critically located lesions. Pentavalent antimony compounds remain the main therapeutic option for all species. They are given intravenously (i.v.), intramuscularly (i.m.), or intralesionally. Cryotherapy, and some systemic antifungal agents have been used successfully. Oral azoles are promising new treatments for lesions caused by L. Major. Several other alternatives and their evidence are also presented. PMID- 12728283 TI - Drug treatments and labeling changes for skin disease introduced in 2002. PMID- 12728284 TI - Etanercept, a TNF antagonist for treatment for psoriatic arthritis and psoriasis. AB - Etanercept (Enbrel, Amgen and Wyeth), a tumor necrosis factor (TNF) antagonist, was approved in January 2002, for the treatment of psoriatic arthritis (PsA). The anti-inflammatory effects of etanercept are due to its ability to bind to the pro inflammatory cytokine TNF, preventing it from interacting with cell-surface receptors and rendering it biologically inactive. Etanercept was evaluated for the treatment of PsA and psoriasis in a preliminary study of 60 patients and in a confirmatory phase III study of 205 patients. In both studies, etanercept was shown to be significantly superior to placebo for the treatment of PsA, evaluated by Psoriatic Arthritis Response Criteria (PsARC) and American College of Rheumatology (ACR) criteria. It also was superior to placebo in improving psoriatic skin lesions, evaluated by the Psoriasis Area and Severity Index (PASI) and target lesion scores. Side-effects were minimal; mild injection site reactions, which resolved during continued therapy, were experienced by approximately one-quarter of the patients. Overall, etanercept is highly effective and well tolerated by patients with PsA, with a safety profile similar to that seen in rheumatoid arthritis. PMID- 12728285 TI - What s new in hair transplants? AB - As hair transplant surgery has evolved, combinations of micrografting and minigrafting have enabled physicians to produce ever more natural combinations of both. A wide range of men and women can now receive significant aesthetic benefits from hair transplants. In recent years, practitioners have further refined hair transplant techniques to ensure that the hairs available for transplantation are used most efficiently. Specifically, methods of harvesting hair, preparing grafts, creating recipient sites, and placing grafts are designed to permit the maximum percentage of hairs to survive and thrive after transplantation. Careful planning, close cooperation with the patient, and a staged surgical approach can also result in hair conservation and optimal cosmesis. PMID- 12728286 TI - Periostitis secondary to interleukin-11 (Oprelvekin, Neumega). Treatment for thrombocytopenia in pediatric patients. AB - Interleukin-11 (Oprelvekin, Neumega) is a newly introduced thrombopoietic growth factor that stimulates production, differentiation, and maturation of megakaryocytes and platelets. Reversible periostitis has been reported as the side effect of the drug in primates and in the phase I/II trials. We report our experience with 5 cases of periostitis, occurring in thrombocytopenic children with three non-malignant and two malignant conditions, out of 24 pediatric patients treated with IL-11 at 75 micro g/kg per day for a median of 17 days. The findings were noted in the clavicle or the proximal humerus. Two patients also had forearm and lower-extremity long-bone involvement. All patients had normal bones before IL-11 was given, changes occurred in both non-malignant and malignant diseases, and periostitis disappeared after use of the drug was discontinued. The distribution and appearance of the changes are similar to prostaglandin E1 and hypervitaminosis A. The changes are reversible after termination of treatment and are most noted in younger patients. The exact mechanism is not clear. The detection of periostitis makes it essential for the radiologists to inquire as to what medications patients are receiving. The pediatric doses (75 g/kg/d) are above those recommended for adult patients (50 g/kg/d) and this may account for the pediatric bone changes of periostitis. PMID- 12728287 TI - Electrical impedance scanning-application of this new technique for lymph node evaluation in children. AB - BACKGROUND: Precise assessment of lymph nodes is crucial to the choice of therapy and prediction of outcome in cases of malignancy. Electrical impedance scanning (EIS) is being experimentally investigated for potential use as a diagnostic tool for differentiation of malignant lesions. Malignancies show different electrical properties with changes in conductivity and capacitance that can be analysed by EIS. Using a TransScan TS-2000 (TransScan Medical, Migdal Ha'Emek, Israel, distributed by Siemens-Elema AB, Solna, Sweden), EIS has been used in various studies for the identification of breast cancer as well as for characterisation of superficial lesions. OBJECTIVE: To evaluate the reliability of EIS for classifying lymph nodes in a pediatric population with sonographically suspicious lesions and to prove its accuracy. MATERIALS AND METHODS: The study population consisted of 77 children (42 boys, 35 girls) aged 1.1-17.1 years. All EIS results were compared to either histopathological findings or long-term follow-up investigations. RESULTS: Sensitivity for malignancies using EIS was 75% and specificity was 87%. The negative predictive value was 93% and the positive predictive value was 60%. CONCLUSIONS: This study suggests the potential usefulness of EIS as an additional imaging modality for the differentiation of lymph-node diseases in children. The histopathological spectrum of malignant lymph node transformation in children compared to studies in adults, and the characteristic meltdown in inflammatory or granulomatous transformed nodes, pose challenges to differentiation based on sonographic evaluation, and also to EIS classification. PMID- 12728288 TI - Determination of bleeding risk using genetic markers in patients taking phenprocoumon. AB - BACKGROUND: In patients on oral anticoagulation with warfarin, genetic variations of the cytochrome P 450-CYP2C9 have recently been associated with very low warfarin requirements. Patients needing low doses had an increased risk for bleeding complications. In Germany, phenprocoumon (having a similar metabolic pathway) is the most commonly employed vitamin K antagonist. Treatment is usually monitored by general practitioners (GPs). OBJECTIVES: To determine whether CYP2C9 variant alleles can serve as risk markers in general-practice patients anticoagulated with phenprocoumon. METHODS: All adult anticoagulated patients in 12 teaching general practices and one university outpatient clinic were to be recruited. Blood samples were taken from 185 patients during routine anticoagulation controls and tested for CYP2C9 mutations. Subjects answered a questionnaire concerning bleeding complications, drug intolerance, and personal and family medical history. Phenprocoumon dosages required for stable anticoagulation were recorded. Odds ratios (OR) with 95% confidence intervals (CI) were calculated based on 2-way cross-tabulations and multivariate logistic regression models, t-tests used where appropriate. RESULTS: Bleeding was reported by 19% of the patients, 2.2% of whom had suffered life-threatening bleeding. CYP2C9 variants were carried by 26.3% of 179 patients tested (17.9% *1/*2, 7.8% *1/*3, 0.6% *2/*3). While presence of a *2 allele was not associated with an increased risk (OR 0.35, CI 0.10-1.24), carriers of the rare *3 alleles had a higher risk of bleeding (OR 3.10, CI 1.02-9.40). With regard to bleeding, carrying CYP2C9*3 was highly specific (94%), though sensitivity was low at 17%; post-test probability of bleeding was 40%. CONCLUSIONS: CYP2C9*3 variants are associated with an increased bleeding risk in patients anticoagulated with phenprocoumon. Screening can identify patients with a high risk of bleeding. Appropriate clinical consequences (restricted indication for anticoagulation, careful induction, adjustment of target INR, closer monitoring or self-testing of INR) as well as the cost-effectiveness of screening for variant CYP2C9 with regard to patient outcomes should be subject of further research. PMID- 12728289 TI - Tailored pharmacokinetic dosing allows self-administration and reduces the cost of IV augmentation therapy with human alpha(1)-antitrypsin. AB - OBJECTIVE: Severe alpha(1)-Antitrypsin (AAT) deficiency (PiZZ) predisposes to the development of emphysema. Intravenous augmentation therapy with purified human AAT has been available since 1988. The dosage has varied from 60 mg/kg body weight once weekly to 250 mg/kg once monthly. We have found the dosage of 120 mg/kg every 2 weeks to be the most convenient for the patients. The treatment is very expensive. The objective of this investigation was to study whether tailored pharmacokinetic dosing of human AAT allows self-administration and reduces the total annual dose and cost of intravenous augmentation therapy. METHODS: Five PiZZ individuals receiving purified human AAT at a dose of 120 mg/kg every 2 weeks were included in the study. Three patients had a percutaneous and one patient had a subcutaneous intravenous injection port system. After a 4-week interruption of the treatment an ordinary dose of 120 mg/kg human AAT was infused. Plasma AAT levels were determined preinfusion, postinfusion, and once daily for 10-14 days. The pharmacokinetic parameters of exogenous AAT were calculated for each patient. Based on these, individual dosage schemes were designed by computer simulation. The patients were treated with the new dose twice weekly for 4 weeks, and plasma AAT was determined immediately before the last two infusions. RESULTS: At a dose of 1 or 2 g twice weekly the median annual consumption of human AAT was reduced from 286 to 156 g/patient. The trough plasma AAT level was maintained above 0.70 g/l, which is considered as protective. The three patients who had an implanted percutaneous venous port system learned to administer the treatment by themselves at home. The other two patients were treated at home by the district nurse. CONCLUSIONS: The results of our study indicate that tailored pharmacokinetic dosing of human AAT reduces the total annual dose and cost of IV augmentation therapy. In addition, this dosing facilitates self-administration of AAT and allows treatment at home. PMID- 12728290 TI - Effect of the CYP2D6 genotype on the pharmacokinetics of tropisetron in healthy Korean subjects. AB - OBJECTIVE: To evaluate the effect of the CYP2D6 genotype on the pharmacokinetics of tropisetron in healthy Korean subjects. METHODS: A single 5-mg capsule of tropisetron was administered orally to 13 healthy subjects. Plasma concentrations were determined by validated HPLC procedures and data were analyzed by using noncompartmental linear PK methods. Four alleles, CYP2D6*1, CYP2D6*2 x2, CYP2D6*5, and CYP2D6*10, were identified by PCR. RESULTS: Thirteen subjects, consisting of two homozygous carriers of the wild type allele ( *1/*1), four heterozygous carriers of poor metabolizer (PM)-associated allele (* 1/*10), six homozygous carriers of PM-associated alleles (four with *10/*10 and two with *5/*10), and one carrier of a duplicated allele *1/*2 x2. All tested pharmacokinetic parameters (AUC(inf), AUC(inf)(NL70), Cmax, Cmax(NL70), T(1/2), and Tec) were significantly different among four different genotypic groups. The mean AUCs of carriers with the heterozygous PM-associated allele and the homozygous PM-associated allele were 1.9- and 6.8-higher than those of carriers with the wild type allele, respectively. In contrast, the mean AUC of carriers with a duplicated allele was 0.5-fold lower than that of those carriers with the wild type allele. CONCLUSION: The presence of CYP2D6*5, CYP2D6*10, and CYP2D6*2 x2 has an important impact on the pharmacokinetics of tropisetron, which may influence clinical response to tropisetron therapy. PMID- 12728292 TI - The effect of attentional cueing on conscious awareness of stimulus and response. AB - Attending to a cued location in space leads to faster reaction times when a stimulus is presented there. The reasons for this attentional effect, and its specific locus in the information-processing chain between stimulus and response, remain unclear. One suggestion is that attention speeds the conscious detection of stimuli. Surprisingly, this possibility appears not to have been tested directly. To resolve this question, we asked subjects to make simple responses to lateralised targets that followed either a valid, invalid or neutral cue, and to judge the perceived time of the target onset, or of their response, by delayed report of the position of a clock hand. Our results showed that only a small and non-significant part of the attentional effect is due to delayed conscious awareness of the stimulus. The greater part of the attentional effect is localised either subsequent to conscious detection of stimuli or occurs in a separate, parallel processing stream from that which generates the motor response. PMID- 12728293 TI - Genotyping of single nucleotide polymorphisms by liquid chromatography electrospray ionization mass spectrometry. PMID- 12728291 TI - Two stages in crossmodal saccadic integration: evidence from a visual-auditory focused attention task. AB - Saccadic reaction time (SRT) toward a visual target stimulus was measured under simultaneous presentation of an auditory non-target (accessory stimulus). Horizontal position of the target was varied (25 degrees left and right of fixation) as well as position and intensity of the auditory accessory. SRT was reduced under the presence of the accessory, and it decreased both with increasing intensity of the auditory accessory and with decreasing distance between target and accessory. The absence of a significant interaction between distance and auditory intensity suggests (1) that the intensity of the accessory stimulus has no direct influence on the process of crossmodal integration, and (2) that spatial position and intensity of the accessory are processed in separate stages. This was supported by a probability inequality test showing that the amount of neural coactivation depends on spatial distance but not on auditory intensity. The results are discussed in the framework of a two-stage model assuming separate processing of unimodal and bimodal characteristics of the stimuli. These results are related to several recent neurophysiological findings. PMID- 12728294 TI - Determination of residual pesticides in olive oil by GC-MS and HPLC-MS after extraction by size-exclusion chromatography. AB - This work describes the development of a method for analyzing pesticide residues in olive oil by GC-MS and HPLC-MS. Pesticides were separated from the oily matrix by size-exclusion chromatography. After extraction, 20 pesticides were separated and analyzed by GC-MS and 11 others HPLC-MS in electrospray mode. The development of this method enabled us to identify and quantify the pesticides of interest. PMID- 12728295 TI - Covalent immobilization of proteases and nucleases to poly(methylmethacrylate). AB - The increased popularity of microfabricated devices formed from plastics such as poly(methylmethacrylate) (PMMA) will benefit from approaches adding (bio)chemical functionality to such surfaces. Here, various proteases and nucleases have been covalently immobilized to PMMA surfaces and shown to retain their enzymatic activity as monitored by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Immobilized enzymes yield structural information at a level equivalent to or exceeding that obtained from conventional homogeneous solution-based approaches. Such an approach could be used to expand the functionality of polymer-based microfabricated devices for biological mass spectrometry. PMID- 12728296 TI - Optical sensor systems for bioprocess monitoring. AB - Bioreactors are closed systems in which microorganisms can be cultivated under defined, controllable conditions that can be optimized with regard to viability, reproducibility, and product-oriented productivity. To drive the biochemical reaction network of the biological system through the desired reaction optimally, the complex interactions of the overall system must be understood and controlled. Optical sensors which encompass all analytical methods based on interactions of light with matter are efficient tools to obtain this information. Optical sensors generally offer the advantages of noninvasive, nondestructive, continuous, and simultaneous multianalyte monitoring. However, at this time, no general optical detection system has been developed. Since modern bioprocesses are extremely complex and differ from process to process (e.g., fungal antibiotic production versus mammalian cell cultivation), appropriate analytical systems must be set up from different basic modules, designed to meet the special demands of each particular process. In this minireview, some new applications in bioprocess monitoring of the following optical sensing principles will be discussed: UV spectroscopy, IR spectroscopy, Raman spectroscopy, fluorescence spectroscopy, pulsed terahertz spectroscopy (PTS), optical biosensors, in situ microscope, surface plasmon resonance (SPR), and reflectometric interference spectroscopy (RIF). PMID- 12728298 TI - Comprehensive two-dimensional gas chromatography. PMID- 12728297 TI - Metal speciation in biomolecules. PMID- 12728299 TI - Advances in optical methods for trace gas analysis. PMID- 12728302 TI - Heat-shock response and its contribution to thermotolerance of the nitrogen fixing cyanobacterium Anabaena sp. strain L-31. AB - Compared to Escherichia coli, the nitrogen-fixing soil cyanobacterium Anabaena sp. strain L-31 exhibited significantly superior abilities to survive prolonged and continuous heat stress and recover therefrom. Temperature upshift induced the synthesis of heat-shock proteins of similar molecular mass in the two microbes. However, in Anabaena sp. strain L-31 the heat-shock proteins (particularly the GroEL proteins) were synthesised throughout the stress period, were much more stable and accumulated during heat stress. In contrast, in E. coli the heat-shock proteins were transiently synthesised, quickly turned over and did not accumulate. Nitrogenase activity of Anabaena cells of sp. strain L-31 continuously exposed to heat stress for 7 days rapidly recovered from thermal injury, although growth recovery was delayed. Exposure of E. coli cells to >4.5 h of heat stress resulted in a complete loss of viability and the ability to recover. Marked differences in the synthesis, stability and accumulation of heat shock proteins appear to distinguish these bacteria in their thermotolerance and recovery from heat stress. PMID- 12728301 TI - Cleavage of CD14 and LBP by a protease from Prevotella intermedia. AB - Periodontitis is an inflammatory disease caused by subgingival microorganisms and their components, such as lipopolysaccharide (LPS). Responses of the host to LPS are mediated by CD14 and LPS-binding protein (LBP). In this study, it was determined that proteases from a periodontal pathogen, Prevotella intermedia, cleave CD14 and LBP, and thereby modulate the virulence of LPS. Culture supernatants from two strains of P. intermedia (ATCC 25611 and 25261) cleaved CD14 and LBP in a concentration-dependent manner. Zymographic and molecular mass analysis revealed the presence of a membrane-associated, 170-kDa, monomeric protease. Class-specific inhibitors and stimulators demonstrated that this enzyme is a metal-requiring, thiol-activated, cysteine protease. The protease was stable over a wide range of temperatures (4-56 degrees C) and pH values (4.5-8.5). This enzyme also decreased the expression of interleukin-1beta (IL-1beta)-specific mRNA in the LPS-activated macrophage-like cell lines U937 and THP-1 in a concentration-dependent manner, indicating that it also cleaves membrane associated CD14. Furthermore, addition of soluble CD14 abrogated protease mediated inhibition of IL-1 mRNA expression induced by LPS. The observations suggest that proteolysis of CD14 and LBP by P. intermedia protease might modulate the virulence of LPS at sites of periodontal infections. PMID- 12728300 TI - Elevated prolactin responses to L-tryptophan infusion in medication-free depressed patients. AB - RATIONALE: Several previous neuroendocrine studies have demonstrated reduced 5 HT1A receptor function in major depressive disorder (MDD). However, hypercortisolaemia or previous drug treatment may have been significant confounds. OBJECTIVES: To replicate previous studies in subjects with MDD who had been drug free for at least 8 weeks and to relate the findings to measures of HPA axis function. METHODS: Hormonal responses to l-tryptophan infusion were measured in patients with MDD (n=20) and healthy controls (n=20). Basal salivary cortisol and DHEA were also profiled. RESULTS: No attenuation of 5-HT1A receptor-dependent neuroendocrine responses (growth hormone, prolactin) was observed in patients with MDD. The prolactin response to l-tryptophan was significantly greater in MDD patients than in healthy controls (P=0.008). There was a significant negative correlation between prolactin response and basal salivary cortisol secretion over the 3 days prior to the test. CONCLUSIONS: These data do not support previous findings of reduced 5-HT1A function in MDD and suggest that hypercortisolaemia or psychotropic medication may have accounted for the attenuation. Basal cortisol, DHEA and the cortiso l-DHEA ratio did not differ between patients and controls, and all patients were psychotropic medication-free. The greater prolactin response to l-tryptophan infusion in depressed subjects may be the result of an increase in dopamine receptor sensitivity, secondary to reduced dopamine levels. PMID- 12728303 TI - Analysis of systemic and regional procalcitonin serum levels during liver transplantation. AB - Elevated procalcitonin (PCT) levels are observed early after orthotopic liver transplantation (OLTx). The aim of this study was to evaluate the changes in systemic and regional PCT serum levels from the time of organ harvesting until the early postoperative phase of OLTx ( n=28) and to investigate the prognostic suitability of postoperative changes in PCT level for the outcome of OLTx ( n=61). Only in seven of 28 donors were higher PCT levels found (0.84+/-0.43 ng/ml). During organ preservation, hepatectomy, and in the anhepatic phase, the PCT levels were in the normal range; in 11 of 28 cases hepatic vein PCT levels were higher during graft flush with own blood than the systemic or portal vein samples at the same time (1.27+/-0.43 ng/ml vs 0.16+/-0.26 ng/ml and 0.23+/-0.15 ng/ml, respectively, P<0.02). The elevation of PCT levels began immediately after graft reperfusion (1.04+/-0.77 ng/ml vs 0.27+/-0.22 ng/ml, P<0.001), and the levels at postoperative day 2 were significantly higher in the case of postoperative complications (30.6+/-19.6 ng/ml vs 4.8+/-3.6 ng/ml, P<0.001). PMID- 12728304 TI - Endovascular cooling with heat exchange catheters: a new method to induce and maintain hypothermia. AB - OBJECTIVE: To test the convenience of a new cooling technique with intravenous heat exchange catheters. DESIGN: Retrospective chart review. SETTING: University hospital neurointensive care unit. PATIENTS: Twenty patients with severe subarachnoid hemorrhage Hunt and Hess Grade 3-5 treated with mild hypothermia. INTERVENTIONS: Cooling to reach target body core temperature (33 degrees C-34 degrees C) was induced as quickly as possible in all patients. In the first ten patients (group one) moderate hypothermia was induced and maintained using cooling blankets. In group two, an 8.5F heat exchange catheter was placed central venous and temperature-adjusted normal saline circulated in a closed-loop system entailing two balloons. MEASUREMENTS AND RESULTS: A total of 2,007 values of body core temperature (BCT) were registered every hour. Foley temperature catheters were used for monitoring BCT in the bladder. The time to reach the target BCT and the stability of temperature during hypothermia were compared between the two groups. No specific complications associated with the new cooling device were observed. Time to reach the target temperature in group two was significantly shorter than in group one (190+/-110 and 370+/-220 min) ( P=0.023). In group one significantly more temperature values were out of the target range (127 of 792 values; 16.0%) than in group two (62 of 1,215 values; 5.1%) ( P<0.0001). CONCLUSIONS: The new endovascular cooling technique seems to be superior for rapid induction of hypothermia and maintaining a more stable temperature than the cooling techniques using blankets and ice bags. PMID- 12728306 TI - A large radial artery false aneurysm after repeated arterial punctures, causing compartment syndrome of the forearm. PMID- 12728305 TI - Nicotinamide increases systemic vascular resistance in ovine endotoxemia. AB - OBJECTIVE: The nuclear enzyme Poly(ADP-Ribose)-Polymerase (PARP) has been hypothesized as playing a major role in various forms of inflammation. PARP activation is induced by DNA strand breakage and can result in intracellular energy depletion and, ultimately, cell death. Further, it is thought to influence cardiovascular function and organ failure in endotoxemia. Here, we investigated the effect of the PARP inhibitor nicotinamide on cardiovascular and liver function in healthy and chronically endotoxemic sheep. DESIGN: Prospective controlled trial. SETTING: University research laboratory. SUBJECTS: 12 female adult sheep. INTERVENTIONS: Six healthy sheep, instrumented for chronic study, received nicotinamide intravenously as a bolus of 40 mg/kg followed by a continuous infusion of 10 mg.kg(-1).h(-1); six animals received the vehicle. One hour after bolus application, a continuous infusion of endotoxin ( Salmonella typhosa, 10 ng.kg(-1).min(-1)) was started. Hemodynamic parameters were determined before and during endotoxemia. MEASUREMENTS AND RESULTS: Treatment with nicotinamide resulted in a significantly higher systemic vascular resistance index and lower cardiac index in endotoxemic animals, but not in controls. It also attenuated endotoxin-induced increase in gamma-glutamyl transferase. CONCLUSIONS: The PARP inhibitor nicotinamide attenuates impairment of cardiovascular function during endotoxemia. In addition, PARP activation may be involved in endotoxin-induced liver injury. PMID- 12728308 TI - Haplotypic analysis of the MMP-9 gene in relation to coronary artery disease. AB - Matrix metalloproteinase-9 (MMP-9) plays an important role in the pathogenesis of atherosclerosis, the pathology underlying the majority of coronary artery disease. We previously identified several polymorphisms in the gene encoding MMP 9. In this study we tested the hypothesis that variation in the matrix metalloproteinase-9 gene influences the development of atherosclerosis. Three common polymorphisms, i.e. -1562C>T, R+279Q and +6C>T, were analysed in 1510 white subjects undergoing coronary angiography. Analyses of individual polymorphisms showed that the frequencies of the C/T and T/T genotypes of the 1562C>T polymorphism were significantly higher in patients with coronary stenosis than in those with a normal angiogram. Logistic regression analyses indicated that individuals carrying the -1562T allele had an approx. 1.5-fold higher risk of developing coronary stenosis (OR 1.49, 95% CI 1.039-2.144), which was equivalent to an over 30% reduction in risk of coronary stenosis in individuals not carrying this allele (OR 0.670, 95% CI 0.467-0.963). The three polymorphisms studied were found to be in strong linkage disequilibrium. Haplotype analyses showed that the C-G-C haplotype (-1562C, +279Q and +6C) was associated with a protective effect against atherosclerosis. Individuals carrying this haplotype were at reduced risk of developing coronary stenosis (OR 0.695, 95% CI 0.530.92). Furthermore, the C-G-C haplotype was associated with less severe coronary atherosclerosis, i.e. carriers of this haplotype were at a lower risk of having coronary stenosis in more than one coronary artery (OR 0.796, 95% CI 0.640.99). These data, together with the previous finding that the -1562T allele has a higher transcriptional activity than the -1562C allele, support the notion that genetic variation with an effect on MMP-9 expression influences the development and progression of atherosclerosis. PMID- 12728309 TI - delta13C and water-use efficiency in Australian grasstrees and South African conifers over the last century. AB - Annual or biannual time courses of plant delta13C (delta13C(p)) over the last century (70-100 years) were recorded for leafbases of four grasstrees (Xanthorrhoea preissii) at four sites in mediterranean Australia and wood of four conifers (Widdringtonia cedarbergensis) at two sites in mediterranean South Africa. There was a strong downward trend of 2-5.5(per thousand ) from 1935 to 1940 to the present in the eight plants. Trends were more variable from 1900 to 1940 with plants at two sites of each species showing an upward trend of 1-2.5 per thousand. Accepting that delta13C of the air (delta13C(a)) fell by almost 2 per thousand over the last century, the ratio of leaf intercellular CO2 to atmospheric CO2 (c(i)/c(a)) rose in five plants and remained unchanged in three over that period. Changes in c(i)/c(a) rather than delta13C(a) were more closely correlated with changes in delta13C(p) and accounted for 6.7-71.8% (22.6 c(i)/c(a)) and 28.2-93.3% (delta13C(a)) of the variation in delta13C(p). We doubt that possible changing patterns of rainfall, water availability, temperature, shade, air pollution or clearing for agriculture have contributed to the overall trend for c(i)/c(a) to rise over time. Instead, we provide evidence (concentrations of Fe and Mn in the grasstree leafbases) that decreasing photosynthetic capacity associated with falling nutrient availability due to the reduced occurrence of fire may have contributed to rising c(i)/c(a). Intrinsic water-use efficiency (W(i)) as a function of (c(a)-c(i)) usually increased linearly over the period, with the two exceptions explained by their marked increase in c(i)/c(a). We conclude that grasstrees may provide equivalent delta13C(p )and W(i) data to long-lived conifers and that their interpretation requires a consideration of the causes of variation in both c(i)/c(a )and delta13C(a). PMID- 12728310 TI - Phenological variation as protection against defoliating insects: the case of Quercus robur and Operophtera brumata. AB - Phenological synchrony between budburst and emergence of larvae is critical for the fitness of many spring-feeding insect herbivores. Therefore, large intraspecific variation in timing of budburst of the host may have a negative effect on the herbivore. We studied how asynchrony between emergence of larvae and budburst affects the fitness of Operophtera brumata (Lepidoptera: Geometridae), a major defoliator of Quercus robur, which can adapt to the phenology of a single tree. It is known that, in maturing leaves of Q. robur, accumulation of condensed tannins has a negative effect on growth of O. brumata. However, there is no information about the effect of hydrolysable tannins and other phenolics that are potential antifeedants. In this study, we also analysed changes in secondary chemistry of the foliage of Q. robur and how different compounds are correlated with growth and survival of O. brumata. The effect of asynchrony on O. brumata was studied in rearing experiments. The neonate larvae were incubated without food for different periods of time. The decline in nutritional quality of foliage was estimated by rearing cohorts of larvae with manipulated hatching times on the leaves of ten individual Q. robur trees. For the chemical analysis, the foliage of these trees was sampled at regular intervals. In the absence of foliage, mortality of neonate larvae started to increase exponentially soon after the larvae emerged. If the larvae missed budburst, the decline in nutritional quality of the foliage led to increased mortality and lower body mass (= fecundity). Hydrolysable tannins were not significantly correlated with performance of the larvae. Only condensed tannins were found to correlate negatively with the growth and survival of O. brumata. Certain individual trees were unsuitable hosts for O. brumata because the decline in quality of the foliage was very rapid. Based on regression equations for increasing rate of mortality and decreasing fecundity, we calculated that a relatively small mismatch of +/-30 degree days between budburst and hatching of larvae leads to a 50% decrease in the fitness of O. brumata. Thus, large phenological variation within a Q. robur stand can limit the colonisation of neighbouring trees by dispersing larvae. Furthermore, the hybridisation of moths adapted to phenologically different trees may lead to maladapted phenology of their offspring. PMID- 12728311 TI - Cis-acting variation in the expression of a high proportion of genes in human brain. AB - Much of the genetic component of human phenotypic diversity, including susceptibility to disease, is proposed to be the result of cis-acting influences on gene expression. If this hypothesis is correct, it implies that cis-acting regulatory variation is a common phenomenon. However, direct evidence in support of this view is scarce. We have applied highly quantitative measures of allele specific expression in order to screen an average of 19 informative subjects (range 8-26) for the presence of common cis-acting influences on the expression of 15 genes by using RNA derived from human brain. We found that, in seven of the 15 assayed genes, at least one individual exhibited relative differences in allelic expression of 20% or more and, in one gene (DTNBP1), allelic expression differences exceeded 50%. These results suggest that cis-acting variation in gene expression commonly occurs in native tissue and hence provide empirical support for the hypothesis that this is potentially an important mechanism underlying human phenotypic diversity. PMID- 12728313 TI - The regeneration process of the striated urethral sphincter involves activation of intrinsic satellite cells. AB - The regeneration of adult skeletal muscle is mediated by satellite cells. Classically, these are considered to be somitically derived cells that colonize the limbs during early embryogenesis. The striated urethral sphincter presents specific developmental characteristics that distinguish it from skeletal muscles, such as the non-somitic origin of its precursor cells and the late formation of its myofibers. This prompted us to determine whether the striated urethral sphincter can regenerate after injury by the same mechanism as skeletal muscles. By means of the single myofiber explant culture technique we investigated the presence of satellite cells in the striated urethral sphincter of male mice and evaluated their ability to recapitulate a myogenic program. In addition, a myotoxic substance (notexin) was injected into the sphincter in order to provoke rapid destruction of the myofibers; the regeneration process was studied by means of electrophysiological and histological techniques. Satellite cells expressing pax7 were found attached to the sphincteric myofibers. They proliferated and expressed MyoD, Myf5 and desmin after 2 days in culture. After 10 days, they formed multinucleated myotubes expressing alpha-actinin-2. In vivo, complete recovery of the striated urethral sphincter, as assessed by normalization of muscle strength and of myofiber number and diameter, was observed after 3 weeks, and resulted from the fusion of myogenic cells. These results demonstrate that the striated urethral sphincter can regenerate by means of a myogenic program involving intrinsic satellite cells. The therapeutic implications of this knowledge and the possible origin of the sphincteric satellite cells are discussed. PMID- 12728312 TI - Hyaluronan is not elevated in urine or serum in Hutchinson-Gilford Progeria Syndrome. AB - Elevations in urinary hyaluronan have been used as the principal laboratory indicator for diagnosis of Hutchinson-Gilford Progeria Syndrome (HGPS). Previous reports have provided evidence suggesting that children with HGPS have altered hyaluronan metabolism as indicated by a mean 17-fold increase in urinary hyaluronan over normal values. In addition, adults with Werner's syndrome have elevated urinary hyaluronan and even more prominent elevations in serum hyaluronan over age-matched controls. It is not known whether serum hyaluronan is elevated or whether serum hyaluronan levels correlate with urinary hyaluronan levels in children with HGPS. In a large cohort of 19 HGPS patients, we sought to confirm elevations in urinary hyaluronan concentration, to establish whether serum hyaluronan is elevated, to measure the size of urinary hyaluronan, and to determine whether serum or urine hyaluronidase levels are altered. We have analyzed urinary and serum hyaluronan levels in patients with HGPS and control patients (1) by using an enzyme-linked immunosorbent assay (ELISA)-like method in which sample hyaluronan in solution and hyaluronan in solid phase compete for a solution of biotinylated hyaluronan-binding protein, and (2) by fluorophore assisted carbohydrate electrophoresis. The size of urinary hyaluronan was measured by using Sepharose CL-6B size exclusion chromatography. Serum and urinary hyaluronidases were evaluated quantitatively, by using ELISA, and qualitatively, by using a gel detection method. HGPS patients did not show a significant elevation in either urinary or serum hyaluronan. We detected no difference in the size of urinary hyaluronan between HGPS children and age matched controls. Serum and urinary hyaluronidase levels were not significantly different in normal and HGPS patients. These studies indicate that neither serum nor urinary hyaluronan concentration is a reliable diagnostic or prognostic marker for HGPS and underscore a difference between adult and childhood progerias. PMID- 12728314 TI - Transitional cell papilloma of the penis associated with human papilloma virus infection. Report of two cases. AB - Condylomas are one of the most common human papilloma virus (HPV)-related benign lesions of the male genitourinary tract. Although rarely, HPV has also been detected along the urinary tract, using molecular techniques, in transitional cell neoplasms without microscopic signs of koilocytic atypia. When affecting the urethra, condylomas are usually limited to its third distal portion. However, transitional cell neoplasms of the urethra are exceptional and in most of the cases remain limited to its proximal portion. To the best of our knowledge, 12 cases of transitional cell carcinomas and only one case of typical transitional cell papilloma have been described in the anterior urethra. We report two exceptional cases of typical transitional cell papilloma of the glans of the penis near the fossa navicularis which showed microscopic signs of HPV infection. PMID- 12728315 TI - Mantle cell lymphoma: improved diagnostics using a combined approach of immunohistochemistry and identification of t(11;14)(q13;q32) by polymerase chain reaction and fluorescence in situ hybridization. AB - INTRODUCTION: Mantle cell lymphoma (MCL) is a clinicopathological entity characterized by an aggressive clinical course, morphological features, and overexpression of cyclin D1 due to juxtaposition of the bcl-1 locus (and CCND1 gene coding for the cyclin D1) to the IgH gene. This phenomenon is caused by t(11;14)(q13;q32). The morphological diagnosis of MCL may pose difficulties. Ancillary methods are available to support the diagnosis. PATIENTS AND METHODS: We studied a group of 32 patients with MCL; 24 men and 8 women. The median age at the diagnosis was 64 years. We characterized the investigated group by histology, and to analyze the immunohistochemical (IHC) profile we used a panel of antibodies including anti-cyclin D1. Polymerase chain reaction (PCR) was used to detect the rearrangement of bcl-1/IgH in 26 cases (in 11 patients, the DNA was isolated from frozen tissues or from nucleated cells of bone-marrow aspirate or peripheral blood, in 15 patients we utilized paraffin-embedded material). Dual color fluorescence in situ hybridization (FISH) on interphase nuclei detecting the t(11;14)(q13;q32) was applied in all 32 cases. RESULTS: Cyclin D1 IHC was positive in 29 of 30 cases tested (97%). In six, the result was weak and difficult to rely on to support the diagnosis. PCR revealed the fusion gene in 14 of the 26 cases (54%). The best yield was obtained from fresh and frozen samples (8 of 11 positive). Using FISH, we identified the translocation in all 32 patients, the findings being easily interpretable in 29 patients. In three cases, the intensity of red and green signals was weaker and difficult to read though the co-hybridized signals were identified. The classical pattern of the translocation was observed in 26 patients, while in 3 we found variant patterns suggesting a loss of the V segment of the IgH gene (2x) and a shift in the breakpoint region at chromosome 11 (1x). CONCLUSION: The diagnosis of MCL should be supported by a complex laboratory approach. Interphase FISH seems a useful complementary method to morphology and IHC. It is applicable to various tissues and cells prepared as tissue imprints or histological sections. PMID- 12728316 TI - Aerenchyma formation in roots of maize during sulphate starvation. AB - Young maize ( Zea mays L., Poaceae) plants were grown in a complete, well oxygenated nutrient solution and then deprived of their external source of sulphate. This treatment induced the formation of aerenchyma in roots. In addition to the effect of sulphate starvation on root anatomy, the presence and location of superoxide anions and hydrogen peroxide, and changes in calcium and pH were examined. By day 6 of sulphate deprivation, aerenchyma started to form in the roots of plants and the first aerenchymatous spaces were apparent in the middle of the cortex. S-starvation also induced thickening of the cell walls of the endodermis. Active oxygen species appeared in groups of intact mid-cortex cells. Formation of superoxide anion and hydrogen peroxide was found in degenerating cells of the mid-cortex. Very few nuclei in the cortex of S-starved roots fluoresced, being shrunken and near to the cell wall. By day 12 of S deprivation, a fully developed aerenchyma was apparent and there were only a few 'chains' of cells bridging hypodermis to endodermis and stele of roots. Cell walls of endodermis of S-starved roots increased 68% in thickness. Intensive fluorescence in the cell walls of the endodermal, hypodermal and to a lesser extent of epidermal cells was observed due to the formation of active oxygen species, while there was no fluorescence in the cortical cells. There was a higher Ca concentration in the cells walls of the endodermis and epidermis, compared to the rest of the S-starved root tissues. A higher pH was observed, mainly in the cell walls of the hypodermis and to a lesser extent in the cell walls of the endodermis. Superoxide anion and hydrogen peroxide was found in degenerating cells of the root cortex. There was no fluorescence of nuclei in the cortex of S-starved roots. PMID- 12728317 TI - Cytosolic potassium homeostasis revisited: 42K-tracer analysis in Hordeum vulgare L. reveals set-point variations in [K+]. AB - Current models of potassium acquisition and cytochemical processes in plants assume that potassium concentrations in the cytosol ([K+]cyt) are maintained homeostatically at approximately 100 mM. Here, we use 42K radiotracer data in the model plant species Hordeum vulgare L. (barley) to show that this assumption is incorrect. Our study reveals that [K+]cyt in root cells of intact barley seedlings is held at a minimum of two physiological set points, coinciding with two fundamentally distinct modes of K+ transport, each of which is characterized by a unique network of fluxes to and from the cytosol, and reflects variations in mechanisms and energetics of K+ transport, cytosolic K+ turnover, flux partitioning, and sensitivity to NH4+. Increased external potassium or ammonium concentrations caused a substantial drop in [K+]cyt, as well as a switch from a transport mode dominated by high-affinity, energy-dependent, influx to a mode dominated by channel-mediated fluxes in both directions across the plasma membrane. Our study provides the first subcellular demonstration of the flexibility, rather than strict homeostasis, of cellular K+ maintenance, and of the dynamic interaction between plant membrane fluxes of the two major nutrient cations K+ and NH4+. PMID- 12728318 TI - Overproduced ethylene causes programmed cell death leading to temperature sensitive lethality in hybrid seedlings from the cross Nicotiana suaveolens x N. tabacum. AB - Reproductive isolation mechanisms (RIMs) often become obstacles in crossbreeding. Hybrid lethality is a subtype of RIM but its physiological mechanism remains poorly elucidated. Interspecific hybrids of Nicotiana suaveolens Lehm. x N. tabacum L. cv. Hicks-2 expressed temperature-sensitive lethality. This lethality was induced by programmed cell death (PCD) that was accompanied by the characteristic changes of animal apoptosis in hybrid seedlings at 28 degrees C but not at 36 degrees C. When hybrid seedlings were cultured at 28 degrees C, DNA fragmentation started in the cotyledon, and nuclear fragmentation subsequently progressed with lethal symptoms spreading throughout the seedlings. At 28 degrees C, ethylene production in hybrid seedlings was detectable at a high level compared with the level in parental seedlings. In contrast, the ethylene production rate in hybrid seedlings cultured at 36 degrees C was equal to that in parental seedlings. Treatment with ethylene biosynthetic inhibitors, amino oxyacetic acid and amino-ethoxyvinyl glycine, suppressed lethal symptoms and apoptotic changes, and also prolonged survival of hybrid seedlings. Thus, the increase in the ethylene production rate correlated closely with expression of lethal symptoms and apoptotic changes in hybrid seedlings. From these observations, we conclude that overproduced ethylene acts as an essential factor mediating PCD and subsequent lethality in hybrid seedlings. Furthermore, the present study has provided the first evidence that ethylene is involved in the phenomenon of hybrid lethality. PMID- 12728320 TI - Actin-related defense mechanism to reject penetration attempt by a non-pathogen is maintained in tobacco BY-2 cells. AB - The actin cytoskeleton is a key player in defense responses during early stages of infection by fungal pathogens. To investigate molecular mechanisms of actin related defense responses, a cultured tobacco ( Nicotiana tabacum L.) BY-2 cell system was devised. When conidia were directly deposited on BY-2 cells, neither a pathogen, Erysiphe cichoracearum, nor a non-pathogen, Erysiphe pisi, was able to form appressoria or haustoria on BY-2 cells. On the other hand, conidia of the powdery mildews formed appressoria on BY-2 cells if they were covered with a thin hydrophobic membrane of Formvar. Percentages of appressoria formation of the powdery mildews on the Formvar-covered BY-2 cells were mostly the same as those on leaf epidermal cells. The pathogen successfully penetrated through the membrane into BY-2 cells and formed haustoria, whereas penetration attempts of the non-pathogen were completely rejected by the BY-2 cells similar to attempts on leaf epidermal cells. On the other hand, when BY-2 cells were treated with actin cytoskeleton-depolymerizing agents, cytochalasins, the non-pathogen became able to penetrate and form haustoria in BY-2 cells. Simultaneously, cytochalasin inhibited callose deposition at penetration sites of the non-pathogen. These results demonstrated that the actin cytoskeleton plays an important role in defense mechanisms against fungal penetration, even in the dedifferentiated cultured cells. The newly devised Formvar-covered cultured cell system will be a useful tool for molecular dissection of signal perception and defense mechanisms of plant cells during the early stage of fungal attack. PMID- 12728319 TI - Differential involvement of the IDRS cis-element in the developmental and environmental regulation of the AtFer1 ferritin gene from Arabidopsis. AB - Four different ferritin genes have been identified in Arabidopsis thaliana, namely AtFer1, 2, 3 and 4. AtFer1, which strongly accumulates in leaves treated with excess iron, contains in its promoter an Iron- Dependent Regulatory Sequence (IDRS). The IDRS sequence is responsible for repression of AtFer1 transcription under conditions of low iron supply. Arabidopsis plants transformed with a 1,400 bp AtFer1 promoter, with either a wild-type or a mutated IDRS fused to the beta glucuronidase (GUS) reporter gene, enabled us to analyze the activity of the AtFer1 promoter in different tissues as well as during age-dependent or dark induced senescence. Our results show that IDRS mediates AtFer1 expression during dark-induced senescence while it does not affect AtFer1 expression during age dependent senescence or in young seedlings. Photoinhibition promoted either by high light or chilling temperature, or wounding, does not activate the AtFer1 promoter. In contrast, AtFer2, AtFer3, AtFer4 transcript abundances are increased in response to photoinhibition and AtFer3 transcript abundance is increased upon wounding. Taken together, our results indicate that other cis-elements, different from the IDRS, regulate the territory-specific or developmental expression of AtFer1 gene. Expression of this gene appears insensitive to some of the environmental stresses tested, which instead up-regulate other members of the Arabidopsis ferritin gene family. PMID- 12728321 TI - Linking respiratory rhythm generation to segmentation of the vertebrate hindbrain. PMID- 12728323 TI - An alternative histochemical method to simultaneously demonstrate muscle nuclei and muscle fibre type. AB - We present a modified histochemical method to examine, simultaneously, nuclei and fibre type in human skeletal muscle. The new procedure (Haem-ATPase) is based on two previously used histochemical protocols. Biopsies were obtained from the rectus abdominis muscle of patients undergoing elective abdominal surgery. Fibre type composition, cross-sectional area (CSA) and nuclei to fibre ratio (N:F) were determined from frozen sections of each biopsy. To test the validity of the new method, serial sections of each biopsy were stained separately using the standard and modified methods. No differences were found in fibre type distribution, mean weighted CSA and N:F when comparing the modified method with the standard methods. The Haem-ATPase method was found to shrink fibre size by at least 3% ( P>0.05) compared with the established myosin acid labile method. We propose that this modified technique is suitable for initial examination of both the nuclei and fibre type in the same frozen sections of human skeletal muscle. PMID- 12728324 TI - Physical demands in working life and individual physical capacity. AB - The purpose of this study was to investigate the prevalence of the excess of metabolic level (metabolic demands in work exceeding one-third of the individual's aerobic capacity) of working men and women today and to describe the population whose metabolic level is exceeded. A second aim was to explore how externally assessed metabolic demands match with the physical function and capacity of working men and women in jobs with the lowest and the highest demands. The aerobic power of each individual (94 men and 94 women) was estimated from heart rate and workload in sub-maximal tests from dynamic legwork on a cycle ergometer. Physical activity was assessed using a task-oriented interview technique. Physical function was measured by tests of muscle endurance in arms, abdomen and legs, handgrip pressure, balance and coordination. The calculation of individual metabolic demands during a "typical working day" showed that 27% of the men and 22% of the women exceeded their metabolic level. The results indicate that the physical fitness is low or somewhat low for two-thirds of the 94 men and for more than one-half of the 94 women. Women in the group with the highest job demands had significantly lower muscle endurance in the abdomen and legs and worse coordination than women in the group with the lowest job demands. Metabolic demands in working life today remain high. This is reflected in a mismatch between individual physical capacity and the physical demands of work for 25% of the population. PMID- 12728322 TI - When endoscopic therapy or pharmacotherapy fails to control variceal bleeding: what should be done? Immediate control of bleeding by TIPS? AB - BACKGROUND: Acute variceal bleeding is the major cause of death in patients with chronic liver disease. This justifies the search for a more effective therapy to achieve rapid and definitive hemostasis in every patient. At present, the recommended standard treatment for acute variceal bleeding consists of immediate drug treatment with terlipressin or octreotide together with early endoscopic band ligation or sclerotherapy. In the case of ectopic varices terlipressin and cyanoacrylate embolization (if varices can be reached by endoscope) are in use. FOCUS: The treatment is considered to have failed when bleeding continues or significant bleeding recurs within 48 h. This indicates the need for emergency transjugular intrahepatic portosystemic shunting (TIPS) which has been regarded as rescue treatment of choice when standard treatment fails. Although randomized studies against supportive treatment are lacking, the high efficacy and relatively low mortality after TIPS implantation are convincing. It is reasonable that smaller shunts should be preferred (probably 8 mm in diameter) since most patients have an increased risk of liver failure. To increase the effect of the shunt with respect to acute hemostasis it should be combined with transjugular embolization of the varices. CONCLUSION: Only strict adherence to the definition of failure of standard treatment and a generous indication to the TIPS implantation before multiorgan failure occurs may decrease the high mortality of acute variceal bleeding. PMID- 12728325 TI - Bacterial endocarditis complicating pregnancy: case report and systematic review of the literature. AB - INTRODUCTION: Infectious endocarditis is a rare life-threatening complication of pregnancy. We report a pregnancy complicated by a 3.5-cm infected vegetation of the tricuspid valve initially presenting as unilateral hip pain as well as systematic review of this entity. SOURCES: A MEDLINE review of the English language literature from 1965 to present using the search terms 'endocarditis', 'pregnancy' and 'infection' as well as review of references was performed. RESULTS: Sixty-eight cases of infectious endocarditis complicating pregnancy were identified. The calculated maternal and fetal mortality rates were 22.1% and 14.7% respectively. CONCLUSIONS: With persistent symptomatic lesions, delivery should be considered without regard to measures of fetal lung maturity because of high fetal mortality rates. PMID- 12728326 TI - Clinical outcome and complications of laparoscopic surgery compared with traditional surgery in women with endometrial cancer. AB - INTRODUCTION: The purpose of this study was to evaluate the feasibility, clinical outcome and complications of laparoscopic surgery in women with endometrial cancer and to compare surgical outcome and postoperative early and late complications with results of traditional laparotomy. METHODS: Forty women with endometrial cancer underwent laparoscopic hysterectomy, bilateral salpingo oophorectomy and pelvic lymphadenectomy. Each patient operated by laparoscopy was matched by age, preoperative clinical stage and histology of the endometrial cancer with a patient treated by the same operation but using traditional laparotomy. Half of these patients underwent total pelvic lymphadenectomy and half had pelvic lymph node sampling. The groups were compared in clinical characteristics, surgical outcomes, recoveries and early and late postoperative complications. RESULTS: The patients in the laparoscopy group had less blood loss, more lymph nodes removed, shorter hospital stay but longer operation time than those treated by laparotomy. Only one (2.5%) laparoscopy was converted to laparotomy due to pelvic adhesions. There were no intraoperative complications in either group. Postoperative complications were more common (55.0%) in the laparotomy than in the laparoscopy group (37.5%). Only one major complication (2.5%) occurred among patients undergoing laparoscopy as compared with three (7.5%) major complications in the laparotomy group. Superficial wound infection was the most common (20%) infection in laparotomy patients while vaginal cuff cellulitis occurred in 10% of laparoscopy patients. Late (>42 days) postoperative complications were almost equally frequent (20.0 and 22.5%) in both groups. Lower extremity lymph edema or pelvic lymph cyst was found in 12.5% of all cases. As a result of surgical staging the disease of 6 women (15%) in both groups was upgraded. CONCLUSIONS: Laparoscopic surgery is a viable alternative to traditional surgery in the management of endometrial cancer. The surgical outcome is similar in both cases. In laparoscopic procedures the operation time is longer but the postoperative recovery time shorter than in laparotomy. Severe complications were limited in both groups, while wound infections can be avoided using laparoscopy. PMID- 12728327 TI - Gene alteration of intestinal intraepithelial lymphocytes in response to massive small bowel resection. AB - BACKGROUND: The intestinal adaptive response [increased epithelial cell (EC) proliferation and apoptosis] after massive small bowel resection (SBR) is partially controlled by intraepithelial lymphocytes (IEL). To identify IEL factors contributing to EC adaptation post-SBR we utilized microarray assays. METHODS: Mice underwent a 70% SBR (SBR1w/SBR4w) or sham operation (Sham1w/Sham4w). After 1 or 4 weeks (1w, 4w) small bowel was harvested, and IEL isolated. Determination of the EC-proliferation rate used BrdU incorporation, and of the EC-apoptotic rate used Annexin V staining. Affymetrix system microarrays (12,491 genes) were performed to examine IEL-mRNA expression. Results were considered significant if fold-change (FC) between groups was >2 and P<0.05 (F test), or FC>3 and 0.05> P >0.01, or FC>4 and P>0.05. Significant genes were confirmed by conventional RT-PCR. RESULTS: The SBR EC-proliferation rate increased significantly in both 1w and 4w groups compared to Sham: SBR1w 0.24+/ 0.07 vs. Sham1w 0.12+/-0.02 (P=0.03); SBR4w 0.35+/-0.04 vs. Sham4w 0.19+/-0.02 ( P<0.01). The EC-apoptotic rate was unchanged in the 1w group, but significantly differed from controls after 4 weeks: SBR4w 39.92+/-6.78 vs. Sham4w 12.56+/-6.44 ( P<0.01). Microarray results were analyzed to identify potential growth modifying IEL genes. The following were identified (function in parenthesis; A, apoptosis; P, proliferation): lipocalin 2 (promotes A), angiotensin converting enzyme (increases A), Rap2 interacting protein (reduces A, promotes P), amphiregulin (promotes P) and leucine-rich-alpha2-glycoprotein (promotes A, reduces P). Based on RT-PCR results these genes showed significant changes between groups. The increase in ACE at 1w preceded the observed apoptotic changes. The alterations in lipocalin 2, Rap2 and amphiregulin at 4w coincided with the marked changes in growth and apoptosis in the SBR mice. CONCLUSIONS: IEL undergo temporal changes after SBR. These findings provide profound insight into potential IEL-dependent regulation of EC homeostasis post-SBR. PMID- 12728328 TI - Intussusception and intestinal malrotation in an infant: a case report. AB - An abnormal cecum position is usually found in patients with intestinal malrotation. We report one case with intussusception and intestinal malrotation in a 10-month-old infant. An unusual radiologic imaging feature and also abnormal intussusception mass location are discussed. PMID- 12728329 TI - Visual cues used by ball-rolling dung beetles for orientation. AB - Ball rolling by dung beetles is considered to be a derived behaviour that evolved under pressure for space, and from competitors at the dung pat. Straight-line orientation away from the pat using a celestial cue should be the most successful rolling strategy to move dung to an unknown burial site. We tested this hypothesis in the field and the laboratory by presenting five species of ball rolling beetles with different orientation tasks, involving reaction to obstacles as well as to reflected sunlight and artificial light sources. Beetles were found to consistently orientate along a chosen route, usually in the direction of the sun. Beetles rolling dung balls successfully negotiated barriers and returned to the original path as did beetles falling from ramps, or rotated about a fixed point while rolling a ball. The sun was found to be the main orientation cue, which could be substituted by reflected or artificial light. However, beetles reoriented themselves less accurately in response to lights in the laboratory, than they did to the reflected sun in the field. It is probable that phototactic orientation using the sun, which is widespread amongst arthropods, has been incorporated in the straight-line foraging behaviour that has evolved in ball rolling dung beetles. PMID- 12728331 TI - Limitations and pitfalls of Couinaud's segmentation of the liver in transaxial Imaging. AB - The segmental anatomy of the human liver has become a matter of increasing interest to the radiologist, especially in view of the need for an accurate preoperative localization of focal hepatic lesions. In this review article first an overview of the different classical concepts for delineating segmental and subsegmental anatomy on US, transaxial CT, and MR images is given. Essentially, these procedures are based on Couinaud's concept of three vertical planes that divide the liver into four segments and of a transverse scissura that further subdivides the segments into two subsegments each. In a second part, the limitations of these methods are delineated and discussed with the conclusion that if exact preoperative localization of hepatic lesions is needed, tumor must be located relative to the avascular planes between the different portal territories. PMID- 12728333 TI - Nonlinear simulation of tumor growth. AB - We study solid tumor ( carcinoma) growth in the nonlinear regime using boundary integral simulations. The tumor core is nonnecrotic and no inhibitor chemical species are present. A new formulation of the classical models [18,24,8,3] is developed and it is demonstrated that tumor evolution is described by a reduced set of two dimensionless parameters and is qualitatively unaffected by the number of spatial dimensions. One parameter describes the relative rate of mitosis to the relaxation mechanisms (cell mobility and cell-to-cell adhesion). The other describes the balance between apoptosis (programmed cell-death) and mitosis. Both parameters also include the effect of vascularization. Our analysis and nonlinear simulations reveal that the two new dimensionless groups uniquely subdivide tumor growth into three regimes associated with increasing degrees of vascularization: low (diffusion dominated, e.g., in vitro), moderate and high vascularization, that correspond to the regimes observed in vivo. We demonstrate that critical conditions exist for which the tumor evolves to nontrivial dormant states or grows self-similarly (i.e., shape invariant) in the first two regimes. This leads to the possibility of shape control and of controlling the release of tumor angiogenic factors by restricting the tumor volume-to-surface-area ratio. Away from these critical conditions, evolution may be unstable leading to invasive fingering into the external tissues and to topological transitions such as tumor breakup and reconnection. Interestingly we find that for highly vascularized tumors, while they grow unbounded, their shape always stays compact and invasive fingering does not occur. This is in agreement with recent experimental observations [30] of in vivo tumor growth, and suggests that the invasive growth of highly-vascularized tumors is associated to vascular and elastic anisotropies, which are not included in the model studied here. PMID- 12728332 TI - Classification of calcaneal fractures by spiral computed tomography: implications for surgical treatment. AB - The purpose of this study was to evaluate spiral computed tomography and multislice CT (SCT/MSCT) with multiplanar reconstructions (MPR) in the classification of calcaneal fractures according to a modified CT classification and to quantify fragment displacement to guide surgical treatment. Forty-eight calcaneal fractures were examined by spiral CT (1- to 2-mm slice thickness, pitch=1.5) with multiplanar reconstructions (MPR). Fractures were grouped according to a modified Munich classification scheme, differentiating six categories of fractures by joint involvement, number of fragments in the posterior facet, and the presence and extent of displacement. A qualitative and quantitative statement was made for the presence of clinical relevant displacement of the posterior articular facet (A: >2 mm), widening of the heel (B: crossing fibular reference line), reduction in calcaneal height (C: >10%), and axis shift of the calcaneocuboid angle (D: >10 degrees ). Treatment recommendations resulting from the CT classification were retrospectively compared with the treatment given by examining the patients' files. There were 10 extra-articular and 38 intra-articular fractures; 8 were in class I (extra articular, nondisplaced), 2 in class II (extra-articular, displaced), 1 in class III (intra-articular, nondisplaced), 20 in class IV (two fragments), 9 in class V (three fragments), and 8 in class VI (>4 fragments), one of the latter being uncertain; 34 showed displacement of the articular facet, 35 widening of the heel, 35 reduction in calcaneal height, and 20 a shift of the axis. In 94% of the cases the procedure recommended by the Munich system of classification was followed; there was disagreement in 1 case in class I and 1 in class IV. Spiral CT allowed fracture classification and quantification of relevant displacement of fragments by radiologists. The implemented recommendations for treatment were adopted by surgeons in most cases. PMID- 12728334 TI - Numerical and exact solutions for continuum of alleles models. AB - Two results are presented for problems involving alleles with a continuous range of effects. The first result is a simple yet highly accurate numerical method that determines the equilibrium distribution of allelic effects, moments of this distribution, and the mutational load. The numerical method is explicitly applied to the mutation-selection balance problem of stabilising selection. The second result is an exact solution for the distribution of allelic effects under weak stabilising selection for a particular distribution of mutant effects. The exact solution is shown to yield a distribution of allelic effects that, depending on the mutation rate, interpolates between the "House of Cards" approximation and the Gaussian approximation. The exact solution is also used to test the accuracy of the numerical method. PMID- 12728335 TI - Inferring population history from genealogical trees. AB - Inference about population history from DNA sequence data has become increasingly popular. For human populations, questions about whether a population has been expanding and when expansion began are often the focus of attention. For viral populations, questions about the epidemiological history of a virus, e.g., HIV-1 and Hepatitis C, are often of interest. In this paper I address the following question: Can population history be accurately inferred from single locus DNA data? An idealised world is considered in which the tree relating a sample of n non-recombining and selectively neutral DNA sequences is observed, rather than just the sequences themselves. This approach provides an upper limit to the information that possibly can be extracted from a sample. It is shown, based on Kingman's (1982a) coalescent process, that consistent estimation of parameters describing population history (e.g., a growth rate) cannot be achieved for increasing sample size, n. This is worse than often found for estimators of genetic parameters, e.g., the mutation rate typically converges at rate under the assumption that all historical mutations can be observed in the sample. In addition, various results for the distribution of maximum likelihood estimators are presented. PMID- 12728336 TI - Bifurcation analysis of the fully symmetric language dynamical equation. AB - In this paper, I study a continuous dynamical system that describes language acquisition and communication in a group of individuals. Children inherit from their parents a mechanism to learn their language. This mechanism is constrained by a universal grammar which specifies a restricted set of candidate languages. Language acquisition is not error-free. Children may or may not succeed in acquiring exactly the language of their parents. Individuals talk to each other, and successful communication contributes to biological (or cultural) fitness. I provide a full bifurcation analysis of the case where the parameters are chosen to yield a highly symmetric dynamical system. Populations approach either an incoherent steady state, where many different candidate languages are represented in the population, or a coherent steady state, where the majority of the population speaks a single language. The main result of the paper is a description of how learning reliability affects the stability of these two kinds of equilibria. I rigorously find all fixed points, determine their stabilities, and prove that all populations tend to some fixed point. I also demonstrate that the fixed point representing an incoherent steady state becomes unstable in an S (n)-symmetric transcritical bifurcation as learning becomes more reliable. PMID- 12728337 TI - Allogeneic stem cell transplantation after a fludarabine/busulfan-based reduced intensity conditioning in patients with myelodysplastic syndrome or secondary acute myeloid leukemia. AB - We report the feasibility and efficacy of a fludarabine/busulfan-based dose reduced conditioning regimen followed by stem cell transplantation from related ( n=19) or unrelated HLA-matched donors ( n=18) in 37 patients with myelodysplastic syndrome (MDS) or secondary acute myeloid leukemia (sAML) who were not eligible for a standard myeloablative conditioning regimen. The conditioning regimen consisted of fludarabine (120-180 mg/m(2)), busulfan (8 mg/kg p.o. or 6.4 mg/kg i.v.), and antithymocyte globulin ( n=25). Graft-versus-host disease (GVHD) prophylaxis consisted of cyclosporine ( n=36) and a short course of methotrexate ( n=29) or mycophenolate mofetil ( n=3). The median age of the patients was 55 years (range: 23-72). The reasons to perform a dose-reduced conditioning were reduced performance status ( n=14), age ( n=12), prior autologous ( n=5) or allogeneic ( n=1) transplantation, or prior/active fungal infection ( n=5). Diagnoses at transplantation were refractory anemia (RA) ( n=8), refractory anemia with excess of blasts (RAEB) ( n=6), RAEB in transformation (RAEB-T) ( n=13), chronic myelomonocytic leukemia (CMML) ( n=3), and sAML ( n=7). Stem cell sources were peripheral blood stem cells (PBSC) ( n=29) or bone marrow ( n=8). One patient received a T-cell-depleted peripheral stem cell graft. Two primary graft failures were observed (6%). Engraftment of leukocytes (>1.0x10(9)/l) and platelets (>20x10(9)/l) was seen after a median of 14 days. Acute GVHD grade II IV was seen in 37%, while severe grade III/IV GVHD was observed in six patients (17%). Chronic GVHD was seen in 13 patients (48%). There were ten deaths (27%) due to treatment (TRM). The probability of TRM was higher in patients with unrelated donors (45 vs 12%, p=0.03) and in patients with poor cytogenetics in comparison to those with a low or intermediate karyotype (75 vs 20%, p=0.009). During follow-up 12 patients relapsed (32%). Patients without chronic GVHD had a significantly higher probability of relapse compared to those with chronic GVHD (70 vs 15%, p=0.02). After a median follow-up of 20 months, the 3-year estimated disease-free survival (DFS) is 38% [95% confidence interval (CI): 21-55%] and the overall survival (OS) is 39% (95% CI: 22-56%). The OS and DFS after related and unrelated transplantations was 45% (95% CI: 19-71%) vs 31% (95% CI: 9-53%) (n.s.) and 51% (95% CI: 29-73%) vs 25% (95% CI: 4-47%) (n.s.), respectively. We conclude that dose-reduced conditioning followed by allogeneic stem cell transplantation from related or unrelated donors is an effective treatment approach in patients with MDS/sAML and might cure a substantial number of patients who are not eligible for a standard allogeneic transplantation. PMID- 12728338 TI - Diagnostics and treatment of pulmonary BALT lymphoma: a report on four cases. AB - Bronchus-associated lymphoid tissue (BALT) is a lymphoid aggregate located in the submucosal area of bronchioles and plays a central role in airway mucosal immunity by inducing the accumulation of secretory IgA-producing cells. Long lasting antigen stimuli promote the hyperplasia of BALT, which may develop into pulmonary mucosa-associated lymphoma (baltoma). Most pulmonary lymphomas are low grade B-cell lymphomas. We have recently treated four patients with BALT lymphoma and this is our first report on their diagnostics and treatment. Based on these cases we wanted to demonstrate the difficulties of differential diagnosis during bronchoscopic and computed tomography (CT) examinations as well as the pitfalls of thoracosurgical vs hemato-oncological treatments. PMID- 12728341 TI - Pseudomass of the sternal manubrium in osteogenesis imperfecta. AB - Skeletal abnormalities such as hypertrophic callus formation and "popcorn" calcifications are rare radiological findings of osteogenesis imperfecta, causing tumor-like appearances on imaging. We report on a 7-year-old girl with osteogenesis imperfecta presenting with hepatomegaly and palpable lymphadenopathy in the left inguinal region on physical examination. Computed tomography examination revealed a high-density mass-like lesion of the manubrium sterni. Ultrasonography and a lateral roentgenogram of the chest verified that this was a pseudomass caused by a bowed sternal manubrium. PMID- 12728342 TI - epsilon-Poly-L-lysine: microbial production, biodegradation and application potential. AB - epsilon-Poly-L-lysine (epsilon-PL) is a homo-poly-amino acid characterized by the peptide bond between the carboxyl and epsilon-amino groups of L-lysine. epsilon PL shows a wide range of antimicrobial activity and is stable at high temperatures and under both acidic and alkaline conditions. The mechanism of the inhibitory effect of epsilon-PL on microbial growth is the electrostatic adsorption to the cell surface of microorganisms on the basis of its poly cationic property. Due to this antimicrobial activity, epsilon-PL is now industrially produced in Japan as a food additive by a fermentation process using Streptomyces albulus. In spite of the practical application of epsilon-PL, the biosynthetic mechanisms of epsilon-PL have not been clarified at all. epsilon-PL producers commonly possess membrane-bound epsilon-PL-degrading aminopeptidase, which might play a role in self-protection. PMID- 12728343 TI - Differential response to low temperature of two Delta6 fatty acid desaturases from Mucor circinelloides. AB - A recently identified Delta6 fatty acid desaturase in Mucor rouxii shows a low sequence homology (approximately 24% at the amino acid level) to that isolated from Mortierella alpina, but is phylogenetically closer to a plant enzyme, suggesting the occurrence of Delta6 desaturase isozymes in Mucorales molds. In the present study, two types of Delta6 desaturases, mcD6-1 ( Mo. alpina type) and mcD6-2 ( M. rouxii type), were cloned from Mucor circinelloides. When the cloned genes were expressed in the yeast Saccharomyces cerevisiae in the presence of a linoleic acid substrate (C18:2Delta9, 12), a newly generated gamma-linolenic acid (C18:3Delta6, 9, 12) was detected in the cells, which confirmed the suspected enzymatic function of the recombinant protein. This is the first report of Delta6 desaturase isozymes present in one organism. Northern analysis demonstrated that the amount of mcD6-2 mRNA was less than half of that of mcD6-1 mRNA in cells grown at 28 degrees C. However, upon cultivation of the cells at 15 degrees C for 0.5-1 h, mcD6-2 mRNA rapidly increased by up to 1.5-fold and then gradually decreased. By contrast, mcD6-1 transcripts levels did not fluctuate significantly for 1 h after the temperature shift, but declined by 75% over the next 2 h. The gamma-linolenic acid content in total fatty acid from M. circinelloides decreased at 28 degrees C, but was maintained at approximately 30% at 15 degrees C. These data suggest that Delta6 desaturase isozymes play physiologically distinct roles in the maintenance of cellular lipids and adaptation to low temperature. PMID- 12728358 TI - The use of prosodic cues in language discrimination tasks by rats. AB - Recent research with cotton-top tamarin monkeys has revealed language discrimination abilities similar to those found in human infants, demonstrating that these perceptual abilities are not unique to humans but are also present in non-human primates. Specifically, tamarins could discriminate forward but not backward sentences of Dutch from Japanese, using both natural and synthesized utterances. The present study was designed as a conceptual replication of the work on tamarins. Results show that rats trained in a discrimination learning task readily discriminate forward, but not backward sentences of Dutch from Japanese; the results are particularly robust for synthetic utterances, a pattern that shows greater parallels with newborns than with tamarins. Our results extend the claims made in the research with tamarins that the capacity to discriminate languages from different rhythmic classes depends on general perceptual abilities that evolved at least as far back as the rodents. PMID- 12728344 TI - A new method for enzymatic preparation of isopentenyladenine-type and trans zeatin-type cytokinins with radioisotope-labeling. AB - We describe a new enzymatic reaction method for the preparation of the radioisotope-labeled cytokinins isopentenyladenine (iP), trans-zeatin (tZ), and their ribosides. The method is based on the three enzyme activities of an adenylate isopentenyltransferase (IPT; EC 2.5.1.27) from Arabidopsis thaliana, an alkaline phosphatase (EC 3.1.3.1) from calf intestine, and a purine-nucleoside phosphorylase (EC 2.4.2.1) from Escherichia coli. The A. thaliana IPT, AtIPT7, utilized both dimethylallyldiphosphate and 4-hydroxy-3-methyl-2-( E)-butenyl diphosphate as isoprenoid donors. The dual specificity of the substrates enabled us to produce iP-type and tZ-type cytokinins separately in the same system simply by switching the substrates. Our method affords a much higher yield of the labeled products than the chemical reaction methods previously used. These labeled compounds will be useful tools for cytokinin research, such as receptor ligand assays and cell metabolism studies. PMID- 12728359 TI - Tindallia californiensis sp. nov., a new anaerobic, haloalkaliphilic, spore forming acetogen isolated from Mono Lake in California. AB - A novel extremely haloalkaliphilic, strictly anaerobic, acetogenic bacterium strain APO was isolated from sediments of the athalassic, meromictic, alkaline Mono Lake in California. The Gram-positive, spore-forming, slightly curved rods with sizes 0.55-0.7x1.7-3.0 microm were motile by a single laterally attached flagellum. Strain APO was mesophilic (range 10-48 degrees C, optimum of 37 degrees C); halophilic (NaCl range 1-20% (w/v) with optimum of 3-5% (w/v), and alkaliphilic (pH range 8.0-10.5, optimum 9.5). The novel isolate required sodium ions in the medium. Strain APO was an organotroph with a fermentative type of metabolism and used the substrates peptone, bacto-tryptone, casamino acid, yeast extract, l-serine, l-lysine, l-histidine, l-arginine, and pyruvate. The new isolate performed the Stickland reaction with the following amino acid pairs: proline + alanine, glycine + alanine, and tryptophan + valine. The main end product of growth was acetate. High activity of CO dehydrogenase and hydrogenase indicated the presence of a homoacetogenic, non-cycling acetyl-CoA pathway. Strain APO was resistant to kanamycin but sensitive to chloramphenicol, tetracycline, and gentamycin. The G+C content of the genomic DNA was 44.4 mol% (by HPLC method). The sequence of the 16S rRNA gene of strain APO possessed 98.2% similarity with the sequence from Tindallia magadiensis Z-7934, but the DNA-DNA hybridization value between these organisms was only 55%. On the basis of these physiological and molecular properties, strain APO is proposed to be a novel species of the genus Tindallia with the name Tindallia californiensis sp. nov., (type strain APO = ATCC BAA-393 = DSM 14871). PMID- 12728360 TI - Metabolism of chloride in halophilic prokaryotes. AB - While much understanding has been achieved on the intracellular sodium and potassium concentrations of halophilic and halotolerant microorganisms and on their regulation, we know little on the metabolism of anions. Archaea of the family Halobacteriaceae contain molar concentrations of chloride, which is pumped into the cells by cotransport with sodium ions and/or using the light-driven primary chloride pump halorhodopsin. Most halophilic and halotolerant representatives of the bacterial domain contain low intracellular ion concentrations, with organic osmotic solutes providing osmotic balance. However, some species show a specific requirement for chloride. In Halobacillus halophilus certain functions, such as growth, endospore germination, motility and flagellar synthesis, and glycine betaine transport are chloride dependent. In this organism the expression of a large number of proteins is chloride regulated. Other moderately halophilic Bacteria such as Halomonas elongata do not show a specific demand for chloride. A very high requirement for chloride was demonstrated in two groups of Bacteria that accumulate inorganic salts intracellularly rather than using organic osmotic solutes: the anaerobic Halanaerobiales and the aerobic extremely halophilic Salinibacter ruber. It is thus becoming increasingly clear that chloride has specific functions in haloadaptation in different groups of halophilic microorganisms. PMID- 12728362 TI - Pain level as a predictor of return to work. PMID- 12728361 TI - Catalysis by methyl-coenzyme M reductase: a theoretical study for heterodisulfide product formation. AB - Hybrid density functional theory has been used to investigate the catalytic mechanism of methyl-coenzyme M reductase (MCR), an essential enzyme in methanogenesis. In a previous study of methane formation, a scheme was suggested involving oxidation of Ni(I) in the starting square-planar coordination to the high-spin Ni(II) form in the CoM-S-Ni(II)F(430) octahedral intermediate. The methyl radical, concomitantly released by methyl-coenzyme M (CoM), is rapidly quenched by hydrogen atom transfer from the coenzyme B (CoB) thiol group, yielding methane as the first product of the reaction. The present investigation primarily concerns the second and final step of the reaction: oxidation of CoB and CoM to the CoB-S-S-CoM heterodisulfide product and reduction of nickel back to the Ni(I) square-planar form. The activation energy for the second step is found to be around 10 kcal/mol, implying that the first step of methane formation with an activation energy of 20 kcal/mol should be rate-limiting. An oxygen of the Gln147 residue, occupying the rear axial position in the oxidized Ni(II) state, is shown to stabilize the intermediate by 6 kcal/mol, thereby slightly decreasing the barrier for the preceding rate-limiting transition state. The mechanism suggested is discussed in the context of available experimental data. An analysis of the flexibility of the F(430) cofactor during the reaction cycle is also given. PMID- 12728364 TI - The lag-effect pattern in the relationship of particulate air pollution to daily mortality in Seoul, Korea. AB - To assess differences in the lag-effect pattern in the relationship between particulate matter less than 10 microm in aerodynamic diameter (PM(10)) and cause specific mortality in Seoul, Korea, from January 1995 to December 1999, we performed a time-series analysis. We used a generalized additive Poisson regression model to control for time trends, temperature, humidity, air pressure, and the day of the week. The PM(10) effect was estimated on the basis of the time series models using the 24-h means and the quadratic distributed-lag models using a cumulative 6-day effect. One interquartile range increase in the 6-day cumulative mean of PM(10) (43.12 microg/m(3)) was associated with an increase in non-accidental deaths [3.7%, 95% confidence interval (CI): 2.1, 5.4], respiratory disease (13.9%, 95% CI: 6.8, 21.5), cardiovascular disease (4.4%, 95% CI: -1.0, 9.0), and cerebrovascular disease (6.3%, 95% CI: 2.3, 10.5). We found the following patterns in the disease-specific lag-effect window: respiratory mortality was more affected by air pollution level on the day of death, whereas cardiovascular deaths were more affected by the previous day's air pollution level. Cerebrovascular deaths were simultaneously associated with the air pollution levels of the same day and the previous day. The patterns in the lag effect from the distributed-lag models were similar to those of a series of time series models with 24-h means. These results contribute to our understanding of how exposure to air pollution causes adverse health effects. PMID- 12728365 TI - Hyperoxaluria in patients with primary distal renal tubular acidosis. PMID- 12728366 TI - Evaluation of the effects of vesicoureteral reflux on urine temperature. PMID- 12728367 TI - Neuroendocrine adaptations in renal disease. AB - Chronic renal failure (CRF) disrupts the time-dependent secretion of multiple hormones. The present review focuses on altered pulsatile release of peptide hormones. CRF is marked by impaired tissue actions, disorderly release patterns, and relative [growth hormone (GH)] or absolute [luteinizing hormone (LH)] deficiency of secretion. At the hypothalamo-pituitary level, experimental evidence suggests that CRF reduces the synthesis and/or release of the cognate hypothalamic releasing factors, GHRH and LHRH, and enforces excessive inhibition by somatostatin. Parathyroid hormone (PTH) and insulin are secreted in both basal and pulsatile modes, wherein the latter is putatively coordinated by autonomic innervation. Amplitude and frequency-dependent adaptations of PTH and insulin outflow fail in CRF, as assessed under steady-state conditions and during metabolic drive (i.e., calcium for PTH and glucose for insulin). A common feature in CRF is a diminished mass of hormone released per burst, due in principle to attenuation of feedforward signals and/or accentuation of (unknown) feedback signals. Damping of neuronal control and/or prolonged network response times may contribute to aberrant pulse frequency, disproportionate basal (nonpulsatile) hormone release, and consistent erosion of secretory process regularity in the uremic state. The homeostatic consequences of distorted secretory dynamics, tissue resistance, impaired hormone clearance, and altered mean agonist concentrations are evident in certain therapeutic interventions, such as GH supplementation in CRF. PMID- 12728368 TI - Gitelman syndrome: when will it turn into Gitelman disease? PMID- 12728370 TI - The effect of different levels of peritoneal CO2 pressure on bleeding time of spleen capsule injury. AB - AIMS: The authors, in contrast to similar injuries with open surgery, had observed spontaneous hemostasis of relatively large spleen capsule injuries during laparoscopic surgery. METHODS: Standard spleen injuries were carried out in 5 anesthetized mongrel dogs at different CO2 pressures and in open surgery. Bleeding was checked every minute by wiping around the injury but not removing the clot. Bleeding time was measured until no more oozing was detected. At every pressure level 3-3 measurements were done and analyzed. Parenchymal pressure of the spleen and systemic blood pressure were detected with direct catheter implantation. RESULTS: In open surgery the average bleeding time was 15.2 min; at 5, 15, and 25 mmHg CO2 pressures bleeding times were 12.3, 10.6, and 9.8 min, respectively. The parenchymal pressure of the spleen (5-6 mmHg) rose synchronously with peritoneal pressure, but no significant changes in systemic blood pressure were seen. CONCLUSION: Peritoneal CO2 pressure may counterweight the parenchymal pressure of the spleen, thus helping hemostasis. There seems to be an inverse proportion between peritoneal pressure and bleeding time. In case of spleen capsule injury during laparoscopic surgery, chances for spontaneous hemostasis are much better compared to open surgery. Attention must be paid to maintain adequate peritoneal pressure. PMID- 12728369 TI - Endoscopic antireflux therapy. PMID- 12728371 TI - Port function after laparoscopic adjustable gastric banding for morbid obesity. AB - BACKGROUND: Laparoscopic adjustable gastric banding (LGB) has gained wide popularity, but information on port function is limited. METHODS: In a prospective nonrandomized study, we analyzed port function and related symptoms in 50 consecutive patients with severe obesity. All patients underwent LGP in a five trocar technique. In 11 patients, the port was placed subcutaneously in the subxiphoid region. In 39 patients, the port was implanted in the left upper abdomen. Mean duration of follow-up was 2.8 years. RESULTS: Patients (12 males and 38 females) had an initial body mass index (BMI) of 47.1 kg/m2. Puncturing the subxiphoidal port was without problems in all 11 patients. However, seven women reported pain and inconvenience when wearing a brassiere. Two underwent port reimplantation in the left upper abdomen (one due to infection; one due to pain). Among the 39 patients with abdominal port implantation, nine patients required port correction (two of them twice). The causes were port dislocation (four cases), difficult puncturing (three), tube leakage (three), and infection (one). CONCLUSION: The high number of complications suggests that the port is the Achilles' heel of LGB. Ports at the subxiphoid site were easier to puncture, but frequently caused pain in female patients. PMID- 12728372 TI - Improvement of foot pedals used during surgery based on new ergonomic guidelines. AB - BACKGROUND: This study aims to create new ergonomic guidelines for the design of foot pedals used during surgery. METHODS: Observations in the operating room, a questionnaire among 45 laparoscopic surgeons/residents, an ergonomic literature study, and clog measurements were used to assess the problems occurring during use and to compile new guidelines for foot pedals. Based on these guidelines a new foot pedal was designed and a prototype was manufactured. RESULTS: During the surgical procedure 91% of the subjects occasionally loses contact with the foot pedal, which 56% experience as very annoying. All subjects think that the current foot pedals obstruct their freedom of movement; 75% occasionally hit the wrong switch and 53% experience physical discomfort in their legs and/or feet. Therefore, 93% of the subjects would like to control the diathermy in a different way. The new prototype of a foot pedal was evaluated in a pilot test and proved to be ergonomically better than the currently used foot pedals. CONCLUSION: The new guidelines for foot pedals result in an ergonomic improvement in their design. PMID- 12728373 TI - Impact of hand dominance, gender, and experience with computer games on performance in virtual reality laparoscopy. AB - BACKGROUND: The impact of gender and hand dominance on operative performance may be a subject of prejudice among surgeons, reportedly leading to discrimination and lack of professional promotion. However, very little objective evidence is available yet on the matter. This study was conducted to identify factors that influence surgeons' performance, as measured by a virtual reality computer simulator for laparoscopic surgery. METHODS: This study included 25 surgical residents who had limited experience with laparoscopic surgery, having performed fewer than 10 laparoscopic cholecystectomies. The participants were registered according to their gender, hand dominance, and experience with computer games. All of the participants performed 10 repetitions of the six tasks on the Minimally Invasive Surgical Trainer-Virtual Reality (MIST-VR) within 1 month. Assessment of laparoscopic skills was based on three parameters measured by the simulator: time, errors, and economy of hand movement. RESULTS: Differences in performance existed between the compared groups. Men completed the tasks in less time than women ( p = 0.01, Mann-Whitney test), but there was no statistical difference between the genders in the number of errors and unnecessary movements. Individuals with right hand dominance performed fewer unnecessary movements ( p = 0.045, Mann-Whitney test), and there was a trend toward better results in terms of time and errors among the residence with right hand dominance than among those with left dominance. Users of computer games made fewer errors than nonusers ( p = 0.035, Mann-Whitney test). CONCLUSIONS: The study provides objective evidence of a difference in laparoscopic skills between surgeons differing gender, hand dominance, and computer experience. These results may influence the future development of training program for laparoscopic surgery. They also pose a challenge to individuals responsible for the selection and training of the residents. PMID- 12728374 TI - Lymphocyte subsets and natural killer cell cytotoxicity after laparoscopically assisted resection of rectosigmoid carcinoma. AB - BACKGROUND: Laparoscopically assisted resection of colorectal carcinoma is technically feasible and minimally invasive. Postoperative immunosuppression also may be reduced. This study compared the lymphocyte subsets and natural killer (NK) cell cytotoxicity in patients after laparoscopically assisted resection with those after open resection of rectosigmoid carcinoma. METHODS: In this study, 40 patients with rectosigmoid carcinoma, but no evidence of metastasis, were randomized to receive either laparoscopically assisted or conventional open resection of the tumor. Blood was collected before the operation, then 24 h, 72 h, and 8 days after the operation for studies of lymphocyte subsets and NK cell cytotoxicity. RESULTS: The lymphocyte subsets and NK cell cytotoxicity of both groups showed typical suppression after surgery. The suppression of T cell activation and NK-like T cells was significantly less after laparoscopically assisted resection than in after open resection, whereas the difference in other lymphocyte subsets and NK cell cytotoxicity was not significant. CONCLUSION: This study showed that some cellular components of the immune system are less suppressed after laparoscopically assisted than after conventional open resection of rectosigmoid carcinoma. This may have implications for tumor recurrence and long-term patient survival. PMID- 12728375 TI - Comparison of biomaterials in the early postoperative period. AB - BACKGROUND: Laparoscopic hernia repair necessitates the use of biocompatible materials. A prospective, double-blind study was conducted to compare two different polypropylene meshes. METHODS: The study included 40 men with primary inguinal hernia undergoing transabdominal preperitoneal polypropylene mesh repair. In 20 of these men, a monofile, heavy-weight, rigid mesh was implanted (group A), and in the remaining 20 men, a smooth, heavy-weight variant of polypropylene mesh was implanted (group B). Complications, pain development, inability to work, physical conditions, testicular volume, and blood circulation were documented. RESULTS: Reconvalescence in group A was slower than in group B. In group A, the postoperative values of the visual scales for pain development were higher; inability to work was 7.3 days longer; urologic disorders were worse; activities of everyday life were more reduced; and SF-36 data showed a significant reduction of physical problems ( p < 0.05). CONCLUSIONS: The polypropylene mesh variant seems to be more compatible with the human organism than conventional mesh. Not only the material, but also the structure seems to influence the comfort of the mesh. PMID- 12728376 TI - The use of laparoscopy in abdominal emergencies. AB - BACKGROUND: The purpose of this article is to describe our experience using laparoscopy in the management of emergent and acute abdominal conditions. METHODS: Between March 1997 and November 2001, 277 consecutive minimally invasive procedures were performed for various nontrauma surgical emergencies. The indications for operation were nonspecific abdominal pain in 129 cases (46%), peritonitis in 64 cases (23%), small bowel obstruction in 52 cases (19%), complications after previous surgery or invasive procedures in 24 cases (9%), and sepsis of unknown origin in 8 cases (3%). RESULTS: Laparoscopy obtained a correct diagnosis in 98.6% of the cases. In 207 patients (75%), the procedure was completed laparoscopically. An additional 35 patients (12.5%) required a target incision. The remaining 35 patients (12.5%) underwent formal laparotomy. The morbidity rate was 5.8%. No laparoscopy-related mortality was observed. CONCLUSIONS: For patients with abdominal emergencies, the laparoscopic approach provides diagnostic accuracy and therapeutic options, avoids extensive preoperative studies, averts delays in operative intervention, and appears to reduce morbidity. PMID- 12728377 TI - A prospective, randomized, unicenter study comparing laparoscopic and open treatments of acute appendicitis. AB - BACKGROUND: Appendectomy in the course of acute appendicitis is one of the most frequently performed surgical procedure in general surgery. The aim of this study was to compare the results of laparoscopic and conventional treatments for acute appendicitis in a prospective, randomized, unicenter study. METHODS: The study involved 200 patients treated for acute appendicitis in the Department of General and Vascular Surgery at Ceynowa Hospital in Wejherowo, Poland. RESULTS: The mean operative time for open surgery was 36.99 min. For laparoscopic method the operation was longer, requiring 47.75 min. Suction drainage was applied in 23 patients treated conventionally and 50 patients treated laparoscopically ( p < 0.05). The requirement for analgesia, measured by the number of metamizole ampules, was significantly higher in the conventional group. Pain on postoperative days 2 and 7 measured using a visual analog scale, was significantly more severe for the patients treated conventionally. The hospital stay in both groups did not differ significantly: 5.03 days for the conventional group and 4.71 days for the laparoscopic group. The time until return to work and social activities in the laparoscopic group (15.85 days) and was significantly shorter than in the conventional group (19.65 days). Seven complications occurred in the conventional group (6.7%) and nine (9.4%) in the laparoscopic group. The difference was not statistically significant. No deaths occurred. CONCLUSIONS: On the basis of the conducted study, it may be assumed that laparoscopic appendectomy is a safe procedure, and that postoperative morbidity is comparable with that for a conventional operation. There was less postoperative pain and shorter recovery time after laparoscopic surgery than after the open procedure. PMID- 12728378 TI - Ergonomic problems encountered by the medical team related to products used for minimally invasive surgery. AB - BACKGROUND: The aim of this study is to gain insight into the problems encountered by the medical team related to products used for minimally invasive surgery. METHODS: An inventory was made of the problems encountered during 12 endoscopic operations performed in one city hospital (Eindhoven, the Netherlands). After the observation, a questionnaire was distributed to all medical staff involved. RESULTS: All categories of personnel had physical, perceptional, and cognitive problems, especially surgeons, residents, and the sterile operation nurse. The main causes were the positioning of apparatus and staff, work clothing, and the limited reach of apparatus and/or instruments. Of the questionnaires, 80% were returned: 50% of the medical staff experienced perceptional problems and 63% had physical discomfort during the surgical procedure. CONCLUSION: The diversity of problems observed and/or reported by the staff during minimally invasive surgery decrease the comfort, efficiency, and safety of the operating-room work environment. Therefore, a new design approach is needed for MIS products in order to address the problems that occur with the current equipment. PMID- 12728379 TI - Totally extraperitoneal repair of recurrent inguinal hernia. AB - BACKGROUND: A variety of procedures with substantial differences in results are employed to treat recurrent inguinal hernia. The advantages of totally extraperitoneal patch repair (TEP) are even more evident when it is applied to recurrent compared to primary hernias. To investigate the superiority of this method more closely, we reviewed our results obtained for recurrent inguinal hernias over a period of 2 years. METHODS: We performed a prospective single center study using data obtained in consecutive patients with recurrent inguinal hernia who were operated on in 1997 and 1998. RESULTS: A total of 179 patients with recurrent inguinal hernia were recruited. Overall, 1329 patients with inguinal hernia were treated in the 2-year period, of whom 1270 underwent TEP. The percentage of recurrent hernias was 14%. The average age of the patients was 56 years. The follow-up rate was 87.5%, and the mean follow-up period was 2.3 years. The 154 patients who were followed up underwent a total of 225 hernia repairs, of which 181 were for recurrent hernias. The average operating time was 57 min. In 68% (104/154) of the patients, adhesions, adherent epigastric vessels, or cicatricial changes were found, which resulted in the inadvertant opening of the peritoneum in 26.3% of the patients. All the openings in the peritoneum were closed by endoscopic suturing. Intraoperative complications developed in 4 patients (2.3%), including one injury to the bladder and three cases of bleeding from side branches of the epigastric vessels. The conversion rate was 0%. The sole postoperative complication was treatment requiring hematomas in 7 patients, in 2 of whom reoperation became necessary. In both cases, a diffuse hemorrhage due to a preoperatively undiagnosed coagulation disorder was found. No cases of wound or patch infection were observed. In a patient undergoing both primary and recurrent hernia repair, displacement of a mesh led to a recurrence on the primary hernia side (recurrence rate, 0.4%; re-recurrence rate, 0%). CONCLUSIONS: Although for its definitive management, recurrent hernia requires a reliable operative technique, current data do not support the recommendation of any of the currently available procedures as the gold standard. In a representative patient population with recurrent hernia, we were able to demonstrate that TEP achieves very good results in terms of re-recurrence rate, intraoperative and postoperative complications, and rehabilitation. Prerequisites for the reliable and low-complication application of the method are a high level of standardization of the procedure and an advanced learning curve. PMID- 12728380 TI - Effect of standard vs extended Roux limb length on weight loss outcomes after laparoscopic Roux-en-Y gastric bypass. AB - BACKGROUND: Increasing the length of the Roux limb in open Roux-en-Y gastric bypass (RYGB) effectively increases excess weight loss in superobese patients with a body mass index (BMI) >50 kg/m2. Extending the RYGB limb length for obese patients with a BMI < 50 could produce similar results. The purpose of this study was to compare the outcomes of superobese patients undergoing laparoscopic RYGB with standard (< or =100-cm) with those undergoing the procedure with an extended (150-cm) Roux limb length over 1-year period of follow-up. METHODS: Retrospective data over 2.5 years were reviewed to identify patients with a BMI < 50 who underwent primary laparoscopic RYGB with 1-year follow-up ( n = 58). Forty-five patients (sRYGB group) received limb lengths < or = 100 cm, including 45 cm ( n = 1), 50 cm ( n = 2), 60 cm ( n = 6), 65 cm ( n = 1), 70 cm ( n = 1), 75 cm ( n = 3), and 100 cm ( n = 31). Thirteen patients (eRYGB group) received 150-cm limbs. Postoperative weight loss was compared at 3 weeks, 3 months, 6 months, and 1 year. RESULTS: Comparing the sRYGB vs the eRYGB group (average +/- SD), respectively: There were no significant differences in age (41.5 +/- 11.0 vs 38.0 +/- 11.9 years), preoperative weight (119.2 +/- 11.9 vs 127.8 +/- 12.5 kg), BMI (43.7 +/- 3.0 vs 45.2 +/- 3.5 kg/m2), operative time (167.1 +/- 72.7 vs 156.5 +/- 62.4 min), estimated blood loss (129.9 +/- 101.1 vs 166.8 +/- 127.3 cc), or length of stay (median, 3 vs 3 days; range, 2-18 vs 3-19). Body weight decreased over time in both groups, except in the sRYGB group between 3 and 6 months and 6 and 12 months after surgery and in the eRYGB group between 6 and 12 months. BMI also decreased over time, except in the eRYGB group between 6 and 12 months. Absolute weight loss leveled out between 6 and 12 months in both groups, with no increase after 6 months. Percent of excess weight loss did not increase in the eRYGB group after 6 months. An extended Roux limb did not significantly affect body weight, BMI, absolute weight loss, or precent of excess weight loss at any time point when the two groups were compared. A trend toward an increased proportion of patients with >50% excess weight loss ( p = 0.07) was observed in the extended Roux limb group. CONCLUSIONS: In this series, no difference in weight loss outcome variables were observed up to 1 year after laparoscopic RYGB. Thus, extending Roux limb length from < or =100 cm to 150 cm did not significantly improve weight loss outcome in patients with a BMI < 50 kg/m2. PMID- 12728381 TI - Endoscopic treatment of postoperative colorectal anastomotic strictures. AB - BACKGROUND: The postoperative development of benign colorectal anastomotic stricture remains a frequent and unsolved problem. METHODS: From 1996 until 2002, we analyzed 94 consecutive patients with postoperative colorectal anastomotic stenosis who were treated endoscopically. RESULTS: Sixty-eight patients were initially resected for malignant disease, and 26 patients for benign conditions. Most frequently, hydrostatic balloon dilatation was performed; in selected cases, it was combined with a laser or argon plasma coagulation (APC) incision, or a laser incision only. Dilatation was successful in 59% of patients resected for cancer and 88% resected for a benign condition. Complications developed in 17 patients (benign restenosis, perforation, abscess); they were significantly more frequent after initial cancer resection than after resection for a benign condition ( p < 0.05). CONCLUSION: High success and low complication rates make endoscopic dilatation the treatment of choice to avoid high-risk reoperations in patients with benign anastomotic stricture. The presence of stapler anastomosis, postoperative leakage, and/or radiotherapy does not significantly impede successful endoscopic dilatation. PMID- 12728382 TI - Transanal endoscopic microsurgery in the treatment of select rectal cancers or tumors suspicious for cancer. AB - BACKGROUND: This study describes a personal experience with the use of transanal endoscopic microsurgery to facilitate surgical access, and to determine the ability of this technique to reduce the need for major abdominal procedure and prevent the need for a temporary or permanent colostomy in select patients with known or suspected rectal cancer. METHODS: The subjects of this study were 43 patients with rectal cancer or tumors who had a high likelihood of malignancy. The 24 men and 19 women comprised two groups: patients with known cancer ( n = 16) and patients with tumors suspicious for cancer ( n = 27), six of whom proved to have invasive malignancy. The tumors ranged in size from 1 to 7 cm (average, 3.5 cm). The tumor's inferior level in the rectum ranged from -1 to 21 cm (average, 6.5 cm). Eleven patients known to have rectal cancer were treated with preoperative radiation or chemoradiation. By ordinary standards, 22 patients would have received an abdominal perineal resection; 14 patients would have qualified for abdominal sphincter-preserving operations; and 7 patients were indeterminate. Full-thickness local excisions were disk excisions ( n = 23), hemicircumferential excisions ( n = 19), and sleeve resection ( n = 1). The ages of the patients ranged from 30 to 91 years (average, 66.7 years). RESULTS: In this study, 90% avoided a major abdominal operation (39/43), and 90% avoided an abdominal perineal resection of the patients (20/22). The complications were as follows: mortality ( n = 10), morbidity ( n = 9), minor wound separation ( n = 6), and major wound separation ( n = 3). Two of these complications were rectovaginal fistulas: the one in a 91-year-old patient who presented with a fistula and the other in a 77-year-old patient who presented with a previously irradiated and incompletely excised cancer. A single instance of locally recurrent cancer required an abdominal perineal resection. There were no other recurrences. Overall, three patients required a stoma (7%). CONCLUSION: Transanal endoscopic microsurgery promises to offer a safe and effective option for the selective treatment of patients with rectal cancer after preoperative chemoirradiation, and for the management of tumors suspicious for rectal cancer. Transanal endoscopic microsurgery used selectively can reduce the need for major abdominal surgery and colostomy. PMID- 12728383 TI - Does laparoscopic antireflux surgery prevent the occurrence of transient lower esophageal sphincter relaxation? AB - BACKGROUND: Transient lower esophageal sphincter relaxation (TLESR) is the most common mechanism underlying gastroesophageal reflux disease (GERD), causing 70% to 100% of the reflux episodes in normal subjects and 63% to 74% of the reflux episodes in patients with reflux disease. This study aimed to evaluate the effect of laparoscopic Nissen fundoplication on TLESR in patients with proven GERD. METHODS: We prospectively followed 73 consecutive patients (13 men and 60 women; mean age, 43.7 +/- 1.72 years) with proven diagnosis of GERD and reported TLESRs found during a 40-min esophageal manometric study. These patients had repeat testing 6 months after undergoing laparoscopic Nissen fundoplication. RESULTS: Laparoscopic Nissen fundoplication increased the basal and nadir lower esophageal sphincter (LES) pressure and significantly reduced the number of TLESRs during the manometric study. No patients after surgery exhibited TLESR with nadir less than 2 mmHg. However, 8 of the 73 patients (11%) exhibited TLESR to a nadir exceeding 50% of basal pressure (mean nadir, 5.0 +/- 1.07 mmHg). CONCLUSIONS: The number of TLESRs is reduced significantly by antireflux surgery. Even accounting for increased basal and nadir pressures, the incidence of TLESR is reduced, suggesting that there may be additional mechanisms involved in this process. PMID- 12728384 TI - Surgical outcome in gastro-esophageal reflux disease patients with inadequate response to proton pump inhibitors. AB - Laparoscopic fundoplication (LF) has been shown to be effective in treatment of patients with gastro-esophageal reflux disease (GERD) requiring long-term medical therapy. Its effectiveness in patients with poor response to proton pump inhibitors (PPIs) has been questioned. We prospectively followed 445 patients with proven GERD inadequately controlled on PPI (up to 120 mg/day) and 274 GERD patients with good response to PPIs (20-120 mg/day) after LF. Patients in both groups underwent 24 h pH testing, esophageal manometry, symptom score evaluation, and quality-of-life (QOL) assessment (SF-36) before and at 6 months, 2 years, and 5 years after surgery. LF was associated with a marked improvement in percentage acid reflux, lower esophageal sphincter pressure, and symptom control in both groups of patients; however, the poor responders to PPIs also had a significant improvement in both physical and mental health component of the QOL assessment. Laparoscopic fundoplication provides an excellent symptom control for GERD patients, even those who have responded inadequately to large doses of PPIs. GERD patients who respond poorly to PPI therapy have significantly lowered physical and mental health QOL scores. Laparoscopic fundoplication in this group of patients leads to marked improvement of both components of QOL by 2 years after surgery. PMID- 12728385 TI - Gallbladder duplication successfully removed laparoscopically using endoscopic nasobiliary tube. AB - Laparoscopic cholecystectomy is sometimes difficult due to complicated biliary anatomy including gallbladder duplication, a rare anomaly of the biliary tract. We report a case of duplicated gallbladder successfully removed under laparoscopy using endoscopic nasobiliary (ENB) tube cholangiography. A 61-year-old Japanese woman presented us with right upper abdominal pain. Ultrasonography revealed two cystic structures lying in the gallbladder fossa, and the upper one contained multiple stones. Endoscopic retrograde cholangiography showed two gallbladders, each of which has a cystic duct draining into the common bile duct separately. Laparoscopic cholecystectomy was planned under the preoperative diagnosis of double gallbladder with gallstones in the accessory gallbladder. The ENB tube was inserted just before the operation. Laparoscopic removal of the double gallbladder was successfully done using the ENB tube to identify the biliary tree anatomy and to close the stump of the cystic duct. In this communication, we would like to stress the usefulness of the ENB tube at the time of laparoscopic biliary surgery in patients with biliary anomalies including gallbladder duplication. PMID- 12728386 TI - Retroperitoneoscopic nephroureterectomy of a horseshoe kidney in a child. AB - Retroperitoneoscopic heminephrectomy used to manage a horseshoe kidney in a 15 month-old boy is reported. The surgery was decided after a progressive increase of arterial blood pressure in the patient followed for poor kidney functioning resulting from vesicoureteral reflux in both ureters of the left duplex moiety of a horseshoe kidney. The retroperitoneoscopy was conducted with three trocars: a 10-mm port inserted under direct vision at the extremity of the left 12th rib and two 5-mm operating trocars. Division of the isthmus was performed using an ultrasonic scalpel. Duration of the pneumoretroperitoneum was 115 min. The patient was discharged on postoperative day 4. At 2 years postoperatively, the patient's arterial blood pressure was normal without treatment. The authors conclude that retroperitoneoscopic nephrectomy for horseshoe kidney is safe and feasible in children, offering all the advantages of minimally invasive surgical procedures. PMID- 12728388 TI - Laparoscopic rectopexy for solitary ulcer of rectum syndrome in a child. AB - BACKGROUND: Solitary ulcer of rectum syndrome occurs rarely in children, and the diagnosis may be difficult. Rectopexy has already been reported with good results in adults and children. We report the first laparoscopic rectopexy in children. METHODS: We report the first case of rectopexy performed according to the Orr Loygue technique using a laparoscopic approach in children. Three trocars were used. A polypropylene mesh was sutured on each side of the rectum and fixed to the promontary with a nonabsorbable suture. The operative time was 150 min. RESULTS: The postoperative course was uneventful. The boy was discharged on postoperative day 3. The rectoscopy at 4 months was normal. The rectal bleeding and pain had disappeared by the 18-month follow-up assessment. CONCLUSION: We report a successful laparoscopic rectopexy for solitary ulcer of rectum syndrome in a child. The procedure is simple, safe, and reproductible, giving good results in the reported case. PMID- 12728387 TI - Hemolysis caused by G-6PD deficiency after a difficult and prolonged therapeutic endoscopic retrograde cholangiopancreatography. AB - Endoscopic retrograde cholangiopancreatography (ERCP), together with its substantial therapeutic capabilities, carries a higher potential for complications than other endoscopic procedures. Common major complications specific to pancreaticobiliary instrumentation include pancreatitis, post sphincterotomy hemorrhage, perforation, and cholangitis with or without systemic sepsis. Two patients underwent therapeutic ERCP for recurrent episodes of abdominal pain and elevation of hepatobiliary enzymes. Endoscopic sphincterotomy was difficult and prolonged. The calculi were successfully extracted by sweeping the choledochus with a balloon-tipped catheter or basket in both cases. The patients experienced postprocedure diffuse abdominal pain unassociated with nausea or vomiting. Laboratory data showed normal serum amylase and lipase 2, 6, and 18 h after the end of procedure, a fall in hematocrit level, and an increase of indirect bilirubin and lactic dehydrogenase. The abdominal pain subsided in 4 to 6 h. The hematocrit level remained stable during the next 3 days, and the patients were very well when discharged. Examination of glucose-6-phosphate dehydrogenase (G-6PD) enzyme levels in red cells 20 days later showed complete enzyme deficiency. This report highlights the importance of examining G-6PD deficiency in patients with post-ERCP abdominal pain, normal serum amylase and lipase, and laboratory findings of hemolysis. PMID- 12728389 TI - Laparoscopic reduction of an ileoileal intussusception and resection of an inverted Meckel's diverticulum in an adult. AB - Intussusception occurs commonly in children, but rarely is observed in adults. Whereas the hydrostatic pressure of a contrast enema often proves diagnostic as well as therapeutic in infants and children, resection usually is required for an underlying bowel pathology in older children and adults. Conventionally, the resection is accomplished at laparotomy. We report the case of a 20-year-old woman who presented with diarrhea and vomiting of 1 week duration. She was found unexpectedly to have intussusception on abdominal ultrasonography. The intussusception was laparoscopically reduced, and a segment of the middle small bowel that harbored an inverted Meckel's diverticulum was resected laparoscopically, after which an intracorporeal anastomosis was fashioned. The ileus resolved on postoperative day 4, and the patient was discharged from hospital on postoperative day 5. The role of the laparoscopic approach in the management of intussusception is discussed. PMID- 12728391 TI - Surgical complications of inguinal and abdominal wall hernias. AB - Inguinal and other abdominal wall hernias are some of the most commonly encountered problems in the pediatric population that require surgical intervention. Surgical repair is straightforward in most cases and can be performed on an outpatient basis. Complications are unusual, but potentially devastating. This article focuses specifically on incidence and risk factors of these complications, reviews the currently accepted treatment options, and points out strategies to avoid them. PMID- 12728392 TI - Editor's note: How we do it: repair of recurrent inguinal hernia. PMID- 12728393 TI - Recurrent undescended testes. AB - Failure of primary orchiopexy for undescended testis is a relatively uncommon occurrence. Redo orchiopexy is a technically challenging procedure with a high rate of success. Prevention of recurrent undescended testis can be achieved with the application of the basic surgical principles of (1) extensive proximal cord mobilization, (2) careful inguinal dissection, and (3) secure intrascrotal gonadal fixation without tension. Secondary or redo orchiopexy requires an experienced pediatric subspecialty surgeon (pediatric or urologic), detailed anatomic knowledge, meticulous operative technique, magnification and delicate instrumentation, injury-free cord dissection, full proximal retroperitoneal cord mobilization, and tension-free intrascrotal fixation. En bloc cord dissection, cord transposition below the deep inferior epigastric vessels, and inguinal cordopexy are helpful adjuncts to reoperative orchiopexy. Successful correction of recurrent undescended testis should approach 95%. PMID- 12728394 TI - Recurrent chest wall anomalies. AB - Chest wall anomalies are a heterogeneous group of malformations requiring repair. Recurrence and the need for secondary repair may occur. Congenital anomalies, including bifid sternum, pentalogy of Cantrell, Jeunes's syndrome and Poland's anomaly, rarely recur. Pectus carinatum may recur in the original surgical area or an adjacent area and most often recurs in patients who undergo repair before completion of teenage growth. Pectus excavatum may recur in approximately 5% of patients. Simple recurrence, floating sternum, or Acquired Jeune's syndrome may result. All of these would require reoperation. Each chest wall anomaly recurrence requires an individualized approach to timing and type of repair. Overall excellent results should be obtained for operative repair of recurrences. PMID- 12728396 TI - Recurrent diaphragmatic hernia. AB - Recurrent congenital diaphragmatic hernia (CDH) is a well-known complication, but one that has not been examined systematically. Review of the literature shows a varied incidence as well as a myriad of treatment strategies. Evaluation of the available data is made more difficult by lack of standardized repair techniques and comparable patient groups. As an increasing number of diaphragmatic hernia patients survive the newborn period, a study of the true incidence of recurrent diaphragmatic hernia, its etiologic factors, and the methods of repair is essential to prevent and treat this problem. PMID- 12728395 TI - Reoperative esophageal surgery. AB - Reoperative esophageal surgery can be a very challenging endeavor. Preoperative evaluation, planning and preparation are essential to optimize results. A general reoperative approach and the range of reconstructive options are outlined. Management of specific problems is discussed including stricture, recurrent gastroesophageal reflux, recurrent tracheoesophageal fistula, esophageal interposition, and recurrent achalasia. PMID- 12728397 TI - The failed fundoplication. AB - Fundoplication is the standard surgical approach to gastroesophageal reflux (GER) in a child. Although successful in many patients, there is a significant risk of complications and failure, especially in high-risk patients such as those with certain types of associated anomalies, diffuse motility disorders, chronic pulmonary disease, neurologic impairment, and young infants. Fundoplication failure can take the form of persistent reflux-related symptoms, symptoms that are caused by complications of the surgery, or anatomic problems such a para esophageal hernia or migration of the wrap into the mediastinum. The most effective strategy for treatment of the child undergoing fundoplication is to prevent failure by careful patient selection, individualization of the operation based on the patient's anatomy and physiology, and meticulous attention to the technical details of the operation. Options for the child with a failed fundoplication include medical management, jejunal feeding using a percutaneous tube or a Roux-en-Y jejunostomy, revision of the fundoplication, or esophagogastric dissociation. If the fundoplication is to be revised, the same principles of patient selection, individualization of the operation, and attention to technique must be used to optimize the chance of success. The primary goal in the treatment of GER is to improve quality of life for the patient and the family. PMID- 12728398 TI - Reoperative surgery for anorectal anomalies. AB - Complications occur during the repair of anorectal malformations relatively frequently. Unfortunately, these complications are often preventable. Furthermore, the consequences of these complications are significant. Not only do patients experience unnecessary pain and suffering, but a secondary operation always renders less optimal functional results. A 20-year experience in the care of children with anorectal malformations was retrospectively analyzed. Patients who previously underwent surgical repair at other institutions, and subsequently required secondary surgery by the primary author were evaluated; 334 patients were identified. Reasons for reoperation included fecal incontinence in 77 patients; dehiscence and retraction in 96; recto-genito-urinary fistulae in 55; persistent urogenital sinus in 31 cloaca patients; acquired vaginal atresia in 21; acquired urethral atresia in 9; posterior urethral diverticulum in 20; and overflow pseudo incontinence in 25 patients. Except for fecal incontinence, all other complications are considered preventable. The source of the complications in almost all other settings are technical errors at the time of the primary repair. Recommendations are presented to help prevent these complications, and suggestions are made on how to treat them when they occur. PMID- 12728399 TI - Reoperative surgery for Hirschsprung's disease. AB - Most patients who undergo a pull-through procedure for Hirschsprung's disease typically do well. A small number of patients have persistent stooling problems, often manifesting with either constipation or recurrent enterocolitis. A group of these patients will not respond to conservative medical management and may need to undergo another operative procedure. Reoperative surgery for Hirschsprung's disease typically falls into 2 categories, those who require a myectomy and those who need a redo pull-through. This article reviews the workup, indications, and techniques for these procedures and discusses previously published, as well as the authors' outcome, with these 2 procedures. PMID- 12728400 TI - Reoperative venous access. AB - The maintenance of long-term venous access is critical to the livelihood of children in a variety of clinical situations, especially those who are dependent on parenteral nutrition. Whereas the traditional routes of either peripheral or central venous access are initially adequate, most of these sites eventually succumb to the pitfalls associated with long-term venous access. This review provides a comprehensive and multidisciplinary approach to the management of reoperative venous access with regard to preoperative planning and imaging and specific techniques in interventional radiology and surgery. PMID- 12728402 TI - Overview of non-Hodgkin's lymphoma: biology, staging, and treatment. AB - The Revised European-American Classification of Lymphoid Neoplasms was recently developed to address the shortcomings of the National Cancer Institute Working Formulation and to establish a common classification system that could be used internationally. The Revised European-American Classification of Lymphoid Neoplasms classification is a major improvement over previous systems and has been shown to define "real" clinical entities that can be diagnosed by expert hematopathologists. Important advances also have recently been made in the staging of lymphomas with the development of the International Prognostic Index, which accurately defines prognostic subgroups and can be used to identify patients who may benefit from more aggressive experimental treatment. With respect to treatment, there remains considerable controversy over the most effective options for patients with indolent lymphomas. However, several novel approaches appear very promising, including monoclonal antibody-based therapy, particularly radioimmunotherapy and purine analogues. In patients with aggressive lymphomas, the treatment options are less controversial. Combination chemotherapy remains the standard of care, but relapse with the development of drug resistance continues to be a problem. Recent data suggest that high-dose therapy with stem cell support may be the treatment of choice for patients with relapsed aggressive lymphomas that remain chemosensitive. PMID- 12728403 TI - Radioimmunotherapy for non-Hodgkin's lymphomas: a historical perspective. AB - One of the most promising therapeutic approaches currently under investigation for low-grade non-Hodgkin's lymphoma is the administration of monoclonal antibodies that recognize tumor-associated antigens. These antibodies can be used either in unmodified form or conjugated to cytotoxic drugs, toxins, or radionuclides. To date, the most promising of the immunoconjugates are radiolabeled antibodies. Radioimmunotherapy is an attractive approach because radiolabeled antibodies are effective even in the face of defective host immune effector function, antigen-negative variants, and poor penetration of the antibody into tumors. Iodine-131-and Yttrium-90-conjugated antibodies have shown superior response rates compared with unconjugated antibodies and have produced complete responses in 15% to 40% of treated patients. A variety of treatment approaches are currently being investigated, including administering radiolabeled antibodies in combination with chemotherapy and administering myeloablative doses with stem-cell rescue. This latter strategy has yielded complete remissions in the majority of treated patients, some durable for more than 5 years. PMID- 12728404 TI - A clinical and scientific overview of tositumomab and iodine I 131 tositumomab. AB - The majority of patients with non-Hodgkin's lymphoma (NHL) who respond to conventional chemotherapy will relapse and eventually become refractory to chemotherapy. Each subsequent remission is typically of similar or shorter duration than the last. Recent developments in radioimmunotherapy using monoclonal antibodies to specifically target malignant B cells have yielded promising results in relapsed and refractory NHL patients. The radiolabeled anti CD20 antibody tositumomab and iodine 131 tositumomab (Bexxar; Corixa Corp, South San Francisco, CA and GlaxoSmithKline, Philadelphia, PA) has been shown to be safe and effective in the treatment of patients with relapsed low-grade (indolent) NHL. Objective responses were achieved in 57% to 71% of patients in phase I to phase III trials, and remission durations were significantly longer in the phase III trial compared with the last remission induced by chemotherapy. In addition, tositumomab and iodine I 131 tositumomab was shown to be effective in the subset analysis of patients with transformed low-grade NHL, which is particularly resistant to conventional therapies. The incidence of transient, nonhematologic adverse events was low and mainly mild to moderate in severity. Hematologic toxicity is the major dose-limiting toxicity associated with radioimmunotherapy; however, patient-specific dosimetry maintained hematologic toxicity within predictable, transient, and manageable limits in the phase II and phase III trials of tositumomab and iodine I 131 tositumomab. Although approximately 10% of patients treated with tositumomab and iodine I 131 tositumomab developed human-antimouse antibodies, treatment with tositumomab does not preclude the administration of subsequent chimeric antibody therapies. In conclusion, these studies show that tositumomab and iodine I 131 tositumomab treatment is safe and induces high response rates and durable remissions in heavily pretreated patients with low-grade or transformed low-grade NHL. PMID- 12728405 TI - The clinical importance of dosimetry in radioimmunotherapy with tositumomab and iodine I 131 tositumomab. AB - Radioimmunotherapy (RIT) is a promising emerging therapy for non-Hodgkin's lymphoma and may ultimately prove useful in the treatment of other tumors. The most extensively investigated RIT agent is tositumomab and iodine I 131 tositumomab (Bexxar; Corixa Corp, South San Francisco, CA, and GlaxoSmithKline, Philadelphia, PA) which has been administered to over 1,000 patients during the past 9 years. As with most drugs, there is considerable interpatient variability in the clearance rate (or total body residence time) of radioimmunoconjugates. The clearance rate of iodine I 131 tositumomab in clinical trials has varied by as much as five-fold. The advantage of RIT with iodine-131, which emits both gamma photons and beta particles, is that by scanning it allows for the determination of the patient-specific total body residence time by the administration of a trace-labeled dose of the radionuclide (ie, dosimetric dose). By administration of the dosimetric (trace-labeled) dose, and determination of the patient's residence time (a measure of how long the radionuclide is retained in the body), the therapeutic dose can be precisely adjusted to maximize the therapeutic effect and minimize toxicity. Tositumomab and iodine I 131 tositumomab is a specific therapeutic at two levels: first, it specifically targets the tumor, delivering a log or more radiation to tumor compared with the rest of the body; and second, the administered dose of radioactivity is patient specific. The paradigm of a targeted drug with a patient-specific dose may become more routine as targeted therapies are further developed along with better assays to directly measure drug levels. For the present, whole-body dosimetry is routinely applied for RIT with tositumomab and iodine I 131 tositumomab and has proven to be a reliable method to determine the patient-specific maximally tolerated therapeutic radiation dose to maximize efficacy while minimizing organ and bone marrow toxicity. PMID- 12728406 TI - Establishing an institutional model for the administration of tositumomab and iodine I 131 tositumomab. AB - Radioimmunotherapy with radiolabeled anti-CD20 antibodies is a promising new treatment approach for low-grade non-Hodgkin's lymphoma. However, the administration of radiolabeled antibodies presents some added complexity. At the University of Nebraska Medical Center (Omaha, NE), an institutional model has been developed that ensures the efficient and safe delivery of tositumomab and iodine I 131 tositumomab (Bexxar; Corixa Corp, South San Francisco, CA and GlaxoSmithKline, Philadelphia, PA). An integrated, multidisciplinary treatment team is responsible for managing all aspects of treatment. Using this model, it is possible to administer tositumomab and iodine I 131 tositumomab safely and effectively in the outpatient setting. Patients can usually be released immediately after treatment. Guidelines and instructions for patient release have been developed and validated and are provided herein. These instructions ensure that radiation exposure of family members and caregivers who are exposed to the patient is maintained as low as reasonably achievable and well within regulatory limits. PMID- 12728408 TI - Insulin, food intake, and reward. AB - Over the past three decades, the hormone insulin has been acknowledged to have multiple effects in the brain, and its role has been validated by the identification of receptors and a transport system for insulin in the central nervous system. Much research has focused on the action of insulin to participate in a feedback loop for the regulation of energy balance. Additionally, more recent studies are demonstrating effects of insulin on brain reward pathways; insulin can interact directly with limbic circuitry to decrease the rewarding or reinforcing value of experimental and natural stimuli, including food. These studies are reviewed in the context of current knowledge of insulin action in the brain, and we offer speculation regarding the relevance of this expanded role of insulin in mental disorders. PMID- 12728409 TI - Do neurotensin receptor agonists represent a novel class of antipsychotic drugs? AB - Schizophrenia is one of the major psychiatric disorders for which effective pharmacotherapy has been available for approximately 50 years. Study of the mechanism of action of these antipsychotic drugs (APDs) has largely focused on the mesolimbic dopamine system and in the neurotransmitter systems that regulate it. Modulation of the neurotensin (NT) circuit in the mesolimbic system can underlie the mechanism of action of APDs. Several lines of evidence support this hypothesis, including: (1) association of NT with neural circuits relevant to the pathophysiology of schizophrenia and the therapeutic effects of APDs; (2) prediction of antipsychotic efficacy and side effect liability based on APD effects on the NT system; (3) low concentrations of NT in the cerebrospinal fluid of a subset of patients with schizophrenia and its normalization after associated clinical improvement with APDs; and (4) remarkable behavioral similarities between peripherally administered APDs and central NT administration. For these reasons, drugs that directly modify the activity of NT systems, particularly NT receptor agonists, could plausibly represent a novel class of APDs. PMID- 12728410 TI - Brain-derived neurotrophic factor, behavior, and new directions for the treatment of mental disorders. AB - Brain-derived neurotrophic factor (BDNF), a member of the neurotrophin family of survival-promoting molecules, plays an important role in the growth, development, maintenance, and function of several neuronal systems. The purpose of this article is to point out evidence for the involvement of this molecule in the maintenance of normal cognitive and emotional functioning, and to outline recent developments using BDNF, or the regulation of endogenous BDNF expression, in the treatment of a variety of mental disorders. This article discusses the important role of BDNF in neuronal growth and survival, in modulating neurotransmission, in activity-directed synaptic remodeling, and in adult neurogenesis. We next outline evidence for the involvement of BDNF in complex behaviors such as learning, feeding, locomotion, responses to painful stimuli, and the management of severe stress. Finally, our review focuses on the involvement of BDNF in treatments for clinical depression and other chronic neurodegenerative processes. We discuss the way that current and future treatment development can be guided by our growing understanding of this molecule's actions in the brain and the ways the expression of BDNF can be regulated. PMID- 12728411 TI - Role of corticotropin-releasing factor family peptides and receptors in stress related psychiatric disorders. AB - Corticotropin-releasing factor (CRF) and related neuropeptides such as urocortin are key mediators of stress in the central nervous system. Through two types of G protein-linked receptors, they play important roles in stress and its relationship to a variety of psychiatric illnesses. CRF appears to play an important role in regulating key neural systems involved in controlling mood, anxiety, feeding behavior, and the interactions between stress and drug addiction. Our improved understanding of the actions of CRF and related peptides reveals not only mechanisms by which stress affects behavior, but also new opportunities to intervene in psychiatric disorders related to stress exposure. PMID- 12728412 TI - The role and therapeutic potential of 5-HT-moduline in psychiatry. AB - The endogenous neuropeptide, 5-HT-moduline, selectively and allosterically interacts with 5-HT(1B) receptors. By binding at a site distinct from that bound by 5-HT, 5-HT-moduline induces structural changes in 5-HT(1B) receptors or stabilizes a particular conformation of these receptors. These conformational changes ultimately lead to the prevention of 5-HT binding resulting in desensitization of these receptors and reduction of the serotonergic function. The efficacy of 5-HT(1B) receptor agonists, for example, has been shown to be reduced by this peptide in vitro and behaviorally. In addition, 5-HT-moduline increases 5-HT release, which is regulated by presynaptic 5-HT(1B) autoreceptors. The release of 5-HT-moduline itself is increased after acute restraint stress in rats, whereas deactivation of 5-HT-moduline by specific antibodies in mice prevents the development of anxiety in a classic behavioral model, suggesting a potential role of the peptide in the control of anxiety. It is thus hypothesized that agents inhibiting the effect of 5-HT-moduline could have anxiolytic activity. Because the serotonergic activity is known to play a key role in psychiatric disorders such as depression and anxiety, compounds capable of mimicking or inhibiting the activity of 5-HT-moduline can represent novel antidepressants or anxiolytics. PMID- 12728413 TI - Autologous blood transfusion after total knee arthroplasty: a randomized, prospective study comparing predonated and postoperative salvage blood. AB - A randomized, prospective study of the use of allogeneic blood was performed in a consecutive series of patients who underwent primary total knee arthroplasty (TKA) and had autologous transfusion either from one unit of predonated autologous blood or from postoperative unwashed blood salvage. In this study, 83 patients (88 knees) were included, with 47 knees in the salvage group and 41 in the predonation group. There were no differences between groups in average age, height, and weight, or gender, diagnoses, or anesthesia type. No significant difference was seen between the groups in the prevalence of allogeneic blood transfusion (5% for the predonation group and 0% for salvage group). Postoperative blood salvage was as effective as predonated autologous blood in preventing the risk associated with allogeneic blood after TKA. PMID- 12728414 TI - The use of a constrained acetabular component for recurrent dislocation. AB - The poor results of surgical treatment of chronic instability after total hip arthroplasty (THA) led to the development of a constrained acetabular component. In this study, 87 constrained THAs implanted for recurrent instability were reviewed retrospectively. Eighty-five hips were available for follow-up evaluation, with an average follow-up period of 58 months. These 85 hips were evaluated at a minimum of 3 years. Two recurrent dislocations were seen, caused by dissociation of the liner from the shell. Four acetabular components and 1 femoral component were revised. Overall, a 2.4% dislocation rate and an 8.2% revision rate were seen. The recurrent dislocation rate of 2.4% represents a significant improvement over other methods reported. Repeat dislocation was only seen in dissociation of cemented liners into well-fixed shells. We do not recommend this mode of fixation. PMID- 12728415 TI - Total joint arthroplasty in human immunodeficiency virus-positive patients: an alarming rate of early failure. AB - The results of 21 total joint arthroplasties (13 knees, 8 hips) in 15 patients were reviewed. There were 13 men and 2 women with an average age of 38 years (range, 28 to 69 years). The mean follow-up period was 10.2 years (range, 2 to 23 years). Six patients died within an average of 10.6 years (range, 3 to 18 years) of joint arthroplasty. All patients died of AIDS. Deep infection developed in 6 joints. Knee Society scores improved significantly, but 13 repeat surgeries were required. An alarmingly high rate of complications was found after total knee and total hip arthroplasty in these patients with human immunodeficiency virus (HIV). Physicians and surgeons should be made aware of this high rate of complications after joint arthroplasty in patients with HIV and include a frank discussion of this information with their patients who are contemplating total joint arthroplasty. PMID- 12728416 TI - Survival analysis of primary cemented total knee arthroplasty: which designs last? AB - Both posterior stabilization and metal backing of the tibial component are design changes that could alter the longevity of total knee arthroplasties (TKAs). Survival analysis data from 16 articles (5,950 knees) were combined to compare design features. No difference was found in survival between posterior stabilized implants and implants that were not stabilized or between metal-backed and all polyethylene tibial components. Those all-polyethylene tibial components that were not stabilized showed significantly better survival than metal-backed, nonstabilized tibial components and posterior stabilized, metal-backed components (P<.05) but not posterior stabilized, all-polyethylene components. According to the currently available literature, posterior stabilization or metal backing of the tibial component does not improve the longevity of primary cemented fixed bearing condylar-type TKAs. PMID- 12728417 TI - Extra-large press-fit cups without screws for acetabular revision. AB - This retrospective study reports the 4- to 6-year results of clinical and radiologic follow-up of 60 acetabular revisions using extra-large hemispherical press-fit cups without additional screw fixation. Bone grafts were used in 17 cases. One cup loosened and required re-revision after 12 months. Initial migration was noted in 6 cases, but showed no progression after less than 2 years. In 2 hips that were revised for deep infection, the cups were found incorporated. At final follow-up, 57 hips (95%) were found firmly fixed radiologically. The Harris Hip score improved from a mean of 58.7 points preoperatively to 90.6 points. Provided a high friction coefficient is present, extra-large hemispherical cups offer sufficient stability even in severe cases. Most do not require additional screw fixation. PMID- 12728418 TI - Mortality after total hip and knee arthroplasty in a medium-volume university practice. AB - This study evaluated the mortality after total knee and hip arthroplasties by a single surgeon in a medium-volume university referral practice. For 610 knee arthroplasties, the in-hospital mortality rate was 0.16%, and overall, 2 patients (0.33%) died within the first 90 days. There was no significant difference in mortality at 1 year or 2 years (P =.55) between primary bilateral one-stage and primary unilateral total knee arthroplasties. For 1,108 total hip arthroplasties, the early in-hospital mortality rate was 0.18%, and overall 5 patients (0.45%) died within the first 90 days. For the entire group of arthroplasties, the overall mortality at 1 year and 2 years was significantly less (both P <.05) than the number of expected deaths in the general population. PMID- 12728420 TI - Knee arthroplasty in octogenarians: results at 10 years. AB - The purpose of this study was to assess the long-term results of total knee arthroplasty (TKA) in octogenarians. Between 1976 and 1989, 1 surgeon at our institution performed 110 TKAs in 90 patients who were 80 years of age or older. No patients were lost to follow-up monitoring; all living patients underwent a minimum of 10 years (range, 10-19 years) followup care. There were 5 (3.4%) surgical complications, but at final follow-up examination, no implants had failed because of aseptic loosening. Of all patients, 96% experienced complete pain relief and 91% showed an excellent knee score (> or = 85) at final follow-up examination. Only 14% of patients had an excellent function score. The 88% death rate for our patient population, the fact that only 14% of patients had an excellent function score after surgery, and the lack of aseptic implant failure by the time of final follow-up examination after an average of 8 years with a conventional TKA raise the question of whether using expensive newer-design implants cost-effectiveness is in octogenarians. PMID- 12728419 TI - Knee range of motion after total knee arthroplasty: how important is this as an outcome measure? AB - We investigated the relationship of knee range of motion (ROM) and function in a prospective, observational study of primary total knee arthroplasty (TKA). Preoperative and 12-month data were collected on 684 patients, including knee ROM, Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) pain and function questionnaire scores, patient satisfaction, and perceived improvement in quality of life (QOL). Only modest correlations were found between knee ROM and WOMAC function (r<0.34). At 12 months we found significantly worse WOMAC function scores for patients with <95 degrees flexion compared with patients with > or =95 degrees (mean, 61.9 vs 75.0; P<.0001). In linear regression models, WOMAC pain and function scores at 12 months were both correlates of patient satisfaction and perceived improvement in QOL (standardized beta>3.5; P<.0001), but knee flexion was not. For assessment of these outcomes, WOMAC function appears to be more important than knee flexion. PMID- 12728421 TI - Primary hybrid total hip arthroplasty with a roughened femoral stem: integrity of the stem-cement interface. AB - One hundred and two consecutive cemented femoral stems were evaluated in 92 patients at an average 9-year follow-up and a minimum 5-year follow-up (range, 5 14 years). The stem used was cobalt chromium with a collar, normalization steps, and a roughened surface (Ra 40); the stem was inserted using contemporary cementing techniques. This series demonstrated a femoral component aseptic loosening rate of 2.0% and a femoral component survivorship of 97.2 +/- 2.0% at 10 years. One of 2 failed stems was revised at 95 months for failure at the cement-bone interface. The second failed stem showed failure at the cement-bone interface with incomplete debonding radiographically at 65 months. The remaining femoral components did not demonstrate any evidence of debonding at the stem cement interface. These results compare favorably with other series of cemented femoral stems, as well as with those with a polished surface. PMID- 12728422 TI - The effect of surgical approaches and tourniquet application on patellofemoral tracking in total knee arthroplasty. AB - The purpose of this study was to assess the influences of surgical approaches and tourniquet application on the lateral retinacular tension (LRT). Single-setting bilateral total knee arthroplasty was performed in 10 patients, and the medial parapatellar and midvastus approaches were randomly performed on each knee using tourniquet application. The LRT was measured using the buckle transducer before and after each approach. Next, the tourniquet was deflated and the measurement was repeated. After the parapatellar approach, LRT was significantly decreased. Conversely, no significant change was seen after the midvastus approach with the tourniquet inflated. However, in the midvastus approach, LRT was significantly decreased after tourniquet deflation. We concluded that both the parapatellar and midvastus approaches influence patellar tracking and LRT. PMID- 12728423 TI - Step-by-step measurements of soft tissue balancing during total knee arthroplasty for patients with varus knees. AB - Using a step-by-step procedure, we measured joint gap during surgery using a simple device with a torque meter in 45 osteoarthritis knee joints (43 patients) with varus deformity. The effects of specific cuts or releases of the anatomic portion on joint gaps were investigated. Each cut or release resulted in various increases in the medial gap from 1.2 to 3.8 mm on average. The final gap measurements averaged 24.1 mm medially and 27.6 mm laterally in extension, and 24.6 mm medially and 27.2 mm laterally in flexion. The results of the measurements showed that each step-by-step procedure had a tendency of gap increase. The results led us to measure soft tissue balancing in a step-by-step procedure during total knee arthroplasty. PMID- 12728424 TI - Long-term results with the cementless Alloclassic brand hip arthroplasty system. AB - The first 100 Alloclassic (Centerpulse, Winterthur, Switzerland) hip prostheses implanted at Allgemeines Krankenhaus Linz were examined in this study. All cases involved primary total hip arthroplasty. Of these 100, 75 could be followed up clinically and radiologically. All living patients were contacted by telephone. The follow-up period was at least 10 years. If the endpoint is defined as removal of the prosthesis due to aseptic loosening, the survival rate was 96.9% for the cup and 100% for the stem after 132 months. The average Harris Hip Score was 85.4 points. Radiolucent lines and osteolyses were found primarily in the proximal zones of the stem. PMID- 12728426 TI - Partial anterior trochanteric osteotomy in total hip arthroplasty: surgical technique and preliminary results of 127 cases. AB - The authors describe a new approach to the hip joint arthroplasty performed in 127 cases of total hip arthroplasty without major complication. A small anterior fragment of greater trochanter, maintaining the insertions of the gluteus minimus and vastus lateralis muscles, is detached. The whole insertion of the gluteus medius is preserved intact, providing good prosthetic stability and rapid recovery of abductor power and gait. Three months after surgery, 74% of patients had recovered good abductor strength with a Merle d'Aubigne and Postel score of 17 points. This surgical approach is technically easy to perform and provides good exposure of the hip. The osteotomized fragment is easily reattached using 2 cerclage wires, and upward displacement after operation was rarely seen. PMID- 12728425 TI - The use of motor-evoked potentials to monitor sciatic nerve status during revision total hip arthroplasty. AB - Sciatic nerve palsy is an uncommon complication after total hip arthroplasty (THA) but can cause permanent functional impairment in the extremity. The goal of this study was to identify specific intraoperative maneuvers that may increase the risk of sciatic nerve injury during revision THA. Motor-evoked potentials (MEPs) were used in combination with electromyography (EMG) monitoring during revision THA in 27 consecutive patients to identify intraoperative events that cause conduction abnormalities through the sciatic and peroneal nerves. MEP monitoring required general anesthesia and a neurophysiologist in the operating room throughout the procedure. Significant electrical events occurred, most commonly during acetabular reconstruction. Hip flexion should be avoided during posterior acetabular retraction when using the posterior approach with posterior dislocation. The position of the sciatic nerve should be clearly identified when complex structural acetabular augmentation with allograft is performed during revision THA. MEP monitoring was used as a research tool for this study. However, we do not use MEP monitoring in our revision THA clinical practice. PMID- 12728427 TI - The incidence of cement extrusion from the acetabulum in total hip arthroplasty. AB - Acetabular cement extrusion (CE) is a common finding after total hip arthroplasty, but the incidence is unclear from the literature, and there is no accepted way of measuring it. We report a retrospective observational study of 100 patients (25 consecutive patients from each of 4 consultants, 112 hips) to determine the site and incidence of CE on postoperative radiographs, and to measure its area and the maximum distance of the extruded cement from the edge of the cement mantle. We found that 50 of 112 (44.6%) hips showed evidence of CE, with a 15.2% incidence of CE > 200 mm(2). PMID- 12728428 TI - Regional and systemic prophylaxis with teicoplanin in total knee arthroplasty: a tissue penetration study. AB - Five patients undergoing total knee arthroplasty (TKA) received 800 mg intravenous teicoplanin systemically 2.5 hours before surgery and 15 patients received 200 mg teicoplanin into a foot vein in the leg to be treated. Samples of bone, synovia, subcutaneous tissue, and skin were collected at 20, 40, and 60 minutes after tourniquet inflation and at the end of surgery. None of the study subjects experienced adverse effects, adverse events, or infections during the postoperative and follow-up period. Mean teicoplanin concentration in the collected tissue ranged from 1.52 to 5.81 mg/L after regional prophylaxis and from 0.9 to 2.94 mg/L after systemic prophylaxis. Bone and soft tissue penetration of teicoplanin after regional prophylaxis with 200 mg is at least comparable with that achieved after systemic prophylaxis with 800 mg. Regional prophylaxis in TKA appears to be safe and valuable. Higher dosages of teicoplanin seem to be needed to ensure coverage against coagulase negative staphylococci. PMID- 12728429 TI - The effect of posterolateral reconstruction on range of motion and muscle strength in total hip arthroplasty. AB - Posterolateral reconstruction of the posterior capsule, piriformis tendon, and external rotators in total hip arthroplasty improves the stability of the hip joint. This study was performed to evaluate the effect of this reconstruction on range of motion (ROM) and circumferential muscle strength. We selected 58 limbs of 29 patients. Posterolateral reconstruction was performed in 13 patients (reconstruction group). Posterolateral reconstruction was not performed in 16 patients (nonreconstruction group). No significant differences were seen in preoperative and postoperative ROM between the 2 groups. The reconstruction group had significantly higher abduction muscle strength (P<.0001) and external rotation muscle strength (P<.01) than the nonreconstruction group. Posterolateral reconstruction may be effective in promoting the recovery of abduction and external rotator muscle strength, and it can improve joint stability without limiting ROM. PMID- 12728430 TI - The application potential of sintered beta-dicalcium pyrophosphate in total joint arthroplasty. AB - An in vitro bone cell culture model was used to evaluate the potential application of sintered beta-dicalcium pyrophosphate (SDCP) in arthroplasty surgery. Primary osteoclasts and osteoblasts were cocultured with different sizes of SDCP particles. The changes in cell counts and the synthesis and secretion of alkaline phosphatase, acid phosphatase, and prostaglandin E(2) in response to the SDCP particles were monitored. When bone cells were cultured with SDCP particles smaller than 53 microm, both the osteoblast and osteoclast cell counts decreased significantly. When the SDCP particles were larger than 177 microm, although the osteoblast population increased significantly, the osteoclast population decreased significantly. Simultaneously, the titer of prostaglandin E(2) in the medium and the cytoplasmic prostaglandin E(2) increased significantly. We concluded that SDCP is a potentially useful bioceramic for the prevention of osteoclast-mediated periprosthetic osteolysis. PMID- 12728431 TI - Observations on retrieved glenoid components. AB - To obtain more information on the pattern of damage of prosthetic glenoid components, we analyzed 7 retrieved glenoid components. The consecutive series included 2 standard polyethylene components and 5 highly crystalline polyethylene glenoids (Hylamer; DePuy Dupont Orthopaedics, Warsaw, IN) retrieved 3 to 12 years after implantation. At revision, 4 of 5 Hylamer components were fractured. Common wear patterns were i) deformation and crumbling of the rim, particularly at the inferior hemicircumference, probably caused by direct contact of the humerus with the prosthetic component; ii) roughening (abrasion and scratching) of the adjacent articulating surface; and iii) concentric and congruous wear centered posteriorly. Available glenoid components may cover an excessive sector of the head. This can result in mechanical restriction of glenohumeral motion and abutment of the humerus against the glenoid rim. Abutment may cause major shear forces and therefore cause glenoid loosening. The value of articular surface mismatch is questionable because retrieved glenoids were worn to a conforming joint. PMID- 12728432 TI - A novel femoral intramedullary plug with sliding mechanism. AB - A novel femoral intramedullary plug with a sliding mechanism has been developed and evaluated clinically. The new plug consists of a pair of specially designed components, each shaped like an obliquely cut cylinder. Postoperative plain radiographs of 30 arthroplasties using the plug were examined for cement leakage, plug migration, and radiolucent line formation between the cement and the femoral cortex. Plugging was complete in 22 cases. Leakage of the cement was seen in 4 cases, and migration of the plug was seen in the other 4 cases. Our study showed the efficacy of the plug in occluding the femoral canal completely in 14 of 22 cases. The plug appears to be promising for clinical applications, because it has good biocompatibility and can occlude the femoral canal tightly. PMID- 12728433 TI - An assessment of sleep disturbance in patients before and after total hip arthroplasty. AB - Patients with osteoarthritis commonly complain of sleep disturbance that may be due to pain. Osteoarthritic hip pain is commonly alleviated by total hip arthroplasty (THA). Forty-eight patients waiting for THA completed a sleep questionnaire and were monitored using actigraphy and sleep diaries for 4 to 5 nights, 1 month before and 3 months after surgery. For the group as a whole, significant improvements were seen in subjective and objective measures of sleep after THA. In this study, 75% of participants reported that their sleep was never or rarely disturbed by hip pain after surgery. Actigraphy indicated significant reductions in time in bed and activity during sleep, more efficient sleep and less fragmented (restless) sleep. Differences between patients aged < 65 and > or = 65 years were observed. These findings support the hypothesis that relief from the pain of osteoarthritis as a result of THA improves sleep quality. Improvements in sleep contribute to the improved quality of life and day-to-day functioning seen after THA. PMID- 12728434 TI - Insufficiency subchondral fracture of the femoral head: report of 4 cases and review of the literature. AB - Insufficiency subchondral fractures are nontraumatic flattened lesions in the superolateral area of the femoral head, occurring in healthy adults. These lesions were recently described and are an infrequent cause of acute hip pain. We report on 4 patients who were diagnosed with an insufficiency subchondral fracture. All the patients showed radiographic progression of the lesion after 4 months, and 3 patients required a total hip arthroplasty. The etiology and the natural history of these rare lesions remain to be elucidated. These lesions should be differentiated from osteonecrosis and transient osteoporosis, because treatment and prognosis may differ. PMID- 12728435 TI - Polyethylene-induced pes anserinus bursitis mimicking an infected total knee arthroplasty: a case report and review of the literature. AB - Polyethylene-induced pes anserinus bursitis after total knee arthroplasty (TKA) with the clinical manifestations mimicking an infected TKA is a very rare occurrence. We report such a case in a 74-year-old woman. She developed recurrent draining papules on the anterolateral aspect of the knee because of polyethylene induced chronic pes anserinus bursitis 8 years after TKA. The diagnosis was established with a sinography study and confirmed by histopathologic examination of the surgical specimen. The symptoms were resolved after surgical excision of the lesion. Pes anserinus bursitis should be included in the differential diagnosis in knees with a draining sinus after TKA, and sinogram is considered the best method to establish the diagnosis. PMID- 12728436 TI - Iliopsoas tendon dysfunction as a cause of pain after total hip arthroplasty relieved by surgical release. AB - Pain after total hip arthroplasty (THA) may be due to a number of factors, including dysfunction of the iliopsoas tendon. We report a case of persistent groin pain after THA. The pain was successfully treated using iliopsoas tenotomy rather than revision of any prosthetic component. PMID- 12728438 TI - Dose escalation in locally advanced carcinoma of the prostate. AB - Radiotherapeutic management of advanced prostate cancer is challenging. Several retrospective analyses showed a dose response for local tumor control before the availability of conformal radiation therapy. Attempts to escalate dose without the benefit of modern treatment planning was commonly fraught with high rates of bowel or bladder complications. The advent of image-guided or computed tomography based treatment planning has allowed safe delivery of high-dose radiation therapy in men with prostate cancer with an acceptable rate of side effects and complications. Several prospective clinical trials have been conducted both at single institutions and in the cooperative group setting. Early evidence suggests that patients with high-risk factors such as advanced clinical stage, high initial prostate-specific antigen, or poorly differentiated tumors may benefit from high-dose 3-dimensional conformal radiation therapy with improved biochemical and local tumor control. A published randomized trial with conformal radiation therapy shows that a modest escalation of radiation dose leads to improved biochemical disease-free survival for a select group of patients. A confirmatory trial within the Radiation Therapy Oncology Group is underway to determine if dose escalation will improve overall survival in men without compromising quality of life. PMID- 12728439 TI - The role of high-dose rate brachytherapy in locally advanced prostate cancer. AB - Although the optimal management of patients with locally advanced prostate cancer remains undefined, sufficient clinical data have emerged showing that patients treated with radiation therapy (RT) have a significantly better outcome as the dose to the gland is escalated. What remains unresolved, however, is how to best deliver these higher tumoricidal doses of RT. Conformal high-dose rate brachytherapy (C-HDR BT) is an alternative means of precise dose escalation that offers similar tumoricidal effects as 3-dimensional (3D) conformal external beam radiotherapy (EBRT) with potential additional advantages. By placing HDR afterloading needles directly into the prostate gland under real-time ultrasound guidance, a steep dose gradient between the prostate and adjacent normal tissues can be generated that is unaffected by organ motion and edema or treatment setup uncertainties. The ability to control the amount of time the single radioactive source dwells at each position along the length of each brachytherapy catheter further enhances the conformity of the dose. In addition, recent radiobiologic data on prostate cancer treatment suggest that the alpha/beta ratio for tumor control is similar to (or possibly even smaller) than that for surrounding late responding normal tissues. If true, hypofractionation (as practiced with C-HDR BT combined with EBRT) would be expected to produce tumor control and late sequelae that are at least as good as achieved with conventional fractionation, with the additional possibility that early sequelae might be reduced. Recent data from several groups performing C-HDR BT in patients with locally advanced disease have confirmed these assumptions. Combined with the physical advantages discussed earlier, C-HDR BT as a means of dose escalation should provide similar tumor control as 3D conformal EBRT with the added advantages of reduced treatment times, less acute toxicity, and no additional technological requirements to account and correct for treatment setup uncertainties and organ motion. The issues that remain unresolved with this technique (as with other methods of dose escalation) revolve around the amount of additional dose required to provide optimal tumor control, the role of androgen deprivation in the management of patients with locally advanced disease, and whether the regional lymphatics should be irradiated. PMID- 12728440 TI - The evolving role of pelvic radiation therapy. AB - Whole pelvic radiotherapy (WPRT) is controversial in the management of prostate cancer. The estimation of the risk of pelvic lymph node involvement in prostate cancer patients will identify those who will potentially benefit from WPRT. Nomograms and equations based on pretreatment prostate-specific antigen (PSA), Gleason score, and/or clinical stage allow clinicians to quickly estimate nodal risk. Most of the studies analyzing WPRT, including a randomized trial from the Radiation Therapy Oncology Group (RTOG), were conducted in the pre-PSA era and did not necessarily include patients at high risk for nodal involvement. The addition of hormonal therapy to WPRT has been shown in 4 major prospective randomized trials to improve survival for some subsets of patients. The preliminary results of RTOG 94-13 show the superiority of WPRT over prostate-only radiotherapy (PORT) in high-risk prostate cancer patients receiving hormonal therapy. For most other solid tumors, the regional lymph nodes are routinely treated by some modality, so it is not surprising that WPRT might benefit a subset of high-risk patients. PMID- 12728441 TI - Treatment of prostate cancer with regional lymph node (N1) metastasis. AB - Prostate cancer with pathologically documented regional lymph node positive disease has been associated with a dismal prognosis in the past. Clinical and/or biochemical progression is evident within 5 years in over 50% treated with external-beam radiotherapy (EBRT) alone, radical prostatectomy (RP) alone, or androgen deprivation (AD) alone. By 10 years after treatment, greater than 75% progress and over half succumb to prostate cancer. In contrast, the results with the combination of EBRT + AD or RP + AD have been very promising. Ten-year biochemical progression and overall survival rates are roughly 20% and 70%, respectively, for patients with subclinical lymph node involvement. Patients with a 10-year life expectancy should be treated aggressively with long-term AD combined with either EBRT or RP. PMID- 12728442 TI - Radiation therapy after radical prostatectomy: a review of the issues and options. AB - The role of postoperative radiation therapy after radical prostatectomy is controversial. Radiation can be delivered as an adjuvant therapy in the immediate postoperative period for high-risk patients or as salvage therapy in the setting of a rising prostate-specific antigen. There are important issues that must be addressed when considering radiation therapy after prior prostatectomy. One issue is the determination of whether a patient has local disease amenable to salvage pelvic radiation or whether the patient has occult metastatic disease. In addition, the radiation oncologist must decide if an acceptable dose of radiation therapy can be administered safely to the prostate bed. There are no published randomized clinical trials on the topic of postprostatectomy radiation therapy, although several have completed accrual or are in progress. Based on the available literature, postoperative radiation is a safe option in the patient at high risk for local recurrence based on adverse pathology or clinical features (eg, extensive extracapsular disease, positive margins, high volume Gleason score >7, and so on). Administration of an adequate dose of prostate bed radiation (ie, >64 Gy) in men with these adverse prognostic features appears to effectively reduce prostate-specific antigen (PSA) recurrence rates. The protracted natural history of prostate cancer requires longer follow-up to determine if survival will be ultimately affected by adjuvant or salvage radiation therapy. Some urologists have advised a "wait and watch policy" for high-risk postprostatectomy patients. Administration of radiation therapy is done only if and when the PSA rises. However, data suggest this approach may have limited durability in high risk prostate cancer and could reduce the likelihood of prolonged progression free survival. This review summarizes published retrospective and prospective data to guide decision making in selecting appropriate candidates for postprostatectomy radiation therapy. PMID- 12728443 TI - Hormones and radiation therapy in locally advanced adenocarcinoma of the prostate. AB - Hormonal dependency of prostate cancer was first described in 1895 by White who showed the initial observation between castration and the treatment of prostatic disease in men with bladder outlet obstruction secondary to benign prostatic hypertrophy. Subsequently, in 1940 Huggins and Hodges demonstrated the association between normal and hyperplastic epithelium of canine prostate glands and adequate levels of circulating androgens. This pioneering work led extensive investigation of the role of androgens in both prostatic growth for benign and malignant disease. The goal of this article is to review the role of hormonal manipulation in prostate cancer in general and more specifically to focus on the role of radiation therapy and hormonal manipulation in locally advanced adenocarcinoma of the prostate. The article is divided into sections that outline the role of different types of hormonal manipulation that are currently available along with the early work combining hormonal manipulation and radiation therapy for prostate cancer. Subsequently, a thorough review of the phase III trials performed to evaluate the potential role of hormonal manipulation in addition to radiation therapy in locally advanced adenocarcinoma of the prostate. Included in that discussion is a section on the potential toxicities of hormonal manipulation. Finally, the current recommendations for the use of hormonal manipulation combined with radiation therapy in locally advanced adenocarcinoma of the prostate is described. PMID- 12728444 TI - Chemotherapy for localized, high-risk prostate cancer. AB - Chemotherapy for prostate cancer currently has an established role in the treatment of hormone refractory prostate cancer. In addition, there is strong evidence that androgen ablation in addition to radiotherapy provides a benefit above and beyond radiotherapy alone in patients with a poor prognosis, perhaps because of an effect of androgen ablation on subclinical distant disease. There are currently several clinical trials that are investigating whether the addition of chemotherapy with known efficacy in the hormone-refractory setting can provide a survival advantage when used adjunctly. The role of chemotherapy in the nonmetastatic prostate, hormone-naive prostate cancer patient will be discussed. PMID- 12728445 TI - Brachytherapy for recurrent prostate cancer after radiation therapy. AB - Prostate cancer specialists routinely see patients with recurrent disease after external beam irradiation. Traditionally, only palliative treatments have been offered with hormonal intervention or simple observation. A significant, although as yet uncertain, percentage of these patients will have only locally recurrent cancer and thus are potentially candidates for curative salvage therapy. Permanent brachytherapy with (125)I or (103)Pd has been used in an attempt to eradicate the remaining prostate cancer and prevent the need for additional intervention. It is particularly critical in this population to identify those patients most likely to have distant metastases or who are unlikely to suffer death or morbidity from their local recurrence to avoid potential treatment morbidity in patients unlikely to benefit from any intervention. Review of the literature shows 5-year freedom from second relapse after salvage brachytherapy in approximately 50% of patients, although with careful case selection second relapse free survival rates of up to 83% may be achieved. A schema is presented, based on the available data, suggesting that it may be possible to identify those patients who are most likely to benefit from salvage treatment. These include men with the following: (1) histologically confirmed local recurrence, (2) no clinical or radiologic evidence of distant disease, (3) adequate urinary function (IPSS < 20), (4) age and overall health indicative of >5- to 10-year life expectancy, (5) prolonged disease-free interval (>2 years) from primary radiation therapy, (6) long prostate-specific antigen (PSA) doubling time (>6-9 months), (7) Gleason sum 0.05). There was a significant increase in bacterial growth in animals subjected to an intra- abdominal pressure of 20 mmHg or above (p < 0.001). As a result, we found that bacterial translocation started when the intra abdominal pressure reached a level of 14 mmHg. Patients should be closely monitored for septic complication risks following laparoscopic procedures in which the intra-abdominal pressure exceeds 20 mmHg. PMID- 12728467 TI - Colonic transit time in diabetic patients--comparison with healthy subjects and the effect of autonomic neuropathy. AB - Constipation and the use of laxatives are relatively common in patients with diabetes mellitus. However, the mechanisms responsible for the constipation are unclear. Even though autonomic neuropathy is regarded as one of the important mechanisms of constipation, it requires further clarification. In addition, the colonic function in diabetic patients requires further investigation. The aim of this study was to compare the colonic transit time between patients with diabetes mellitus and healthy subjects, and correlate it to the presence of cardiovascular autonomic neuropathy. The colonic transit time was measured by a noninvasive, radio-opaque marker method, and the presence of cardiovascular autonomic neuropathy was evaluated by the beat-to-beat variation and the orthostatic hypotension. Constipation was defined by the Rome II criteria. The mean total colonic transit time of the 28 diabetic patients (34.9 +/- 29.6 h, mean +/- S.D.) was significantly longer than that of the 28 healthy subjects (20.4 +/- 15.6 h, p < 0.05). Among the diabetic patients, 9/28 (32%) had constipation and 14/28 (50%) had cardiovascular autonomic neuropathy. The diabetic patients with constipation showed longer total, left and recto-sigmoid colonic transit times than those without constipation. However, the mean colonic transit time of diabetic patients with and those without cardiovascular autonomic neuropathy was similar. In conclusion, other mechanisms than the mere presence of cardiovascular autonomic neuropathy might be more relevant to the development of constipation in patients with diabetes mellitus. PMID- 12728468 TI - Simple retrograde balloon dilation for treatment of ureteral strictures: etiology based analysis. AB - This study consists of an analysis of the results of simple retrograde balloon dilation in the treatment of ureteral strictures for the evaluation of the long term efficacy of this procedure, and the factors affecting its success rate. A prospective study was performed on 43 ureteral strictures (22 malignant, 21 benign) from 37 patients treated with retrograde balloon dilation from October 1997 to May 1999. After the stricture segments were dilated, ureteral stents were indwelled uniformly for 3 weeks. Strictures were postoperatively followed up radiographically at 1, 3, 6 and 12 months, and annually thereafter. Success was defined by symptomatic and radiographic improvement. The follow-up periods ranged from 8 to 57 months (mean 41 months). The success rates of the benign strictures at 12 and 36 months were much higher than those of the malignant strictures (67 and 57% vs., 18 and 14%, p=0.0009). While 56% and 47% of the patients with strictures shorter than 2 cm were successful at 12 and 36 months, respectively, none with strictures longer than 2 cm were successful at the same follow-up periods (p=0.0002). Of the successful benign cases with a shorter segment at 12 months, 12 out of 14 (86%) showed persistent long-term successes at the 36 months follow-up. Other prognostic factors, such as sex, age, location, disappearance of a waist, dilation time and grade of hydronephrosis, were not found to influence the success rate. A multivariate analysis revealed the etiology and stricture length were the only significant prognostic factors affecting the final outcome (p=0.030 and p=0.0262, respectively, by Cox's proportional hazards model). In consideration of its minimal invasiveness and acceptable long-term outcome, simple retrograde balloon dilation is an effective treatment modality for benign ureteral stricture with a short segment (< or = 2 cm), and a shorter duration of stenting (3-weeks) is viable. PMID- 12728469 TI - Relationship between urinary endogenous steroid metabolites and lower urinary tract function in postmenopausal women. AB - To investigate the relationship between the endogenous steroid hormones and the lower urinary tract function in postmenopausal women. Thirty postmenopausal volunteer women who did not have lower urinary tract symptoms or hormone replacement therapy were enrolled in this study. Urodynamic studies included uroflowmetry, multi-channel cystometry, and urethral pressure profilometry were conducted. Gas Chromatography- Mass Spectroscopy(GC-MS) was used to measure the urinary endogenous steroid hormone metabolites. The relationship between the urinary profile of the endogenous steroids and the urodynamic parameters of these patients were investigated. The mean ages of the patients were 60.6 +/- 5.5 years, and the Body Mass Index (BMI) averaged 24.56 +/- 2.23 (kg/m2). Of the progesterone metabolites, pregnandiol was significantly related to the residual volume in the uroflowmetry and the functional urethral length parameters (R=0.98, p=0.000; R= -0.65, p=0.04). Pregnantriol was significantly related to the maximum flow rate, the residual volume in uroflowmetry, the maximum urethral closure pressure and the functional urethral length (R=-0.64, p=0.04; R=0.82, p=0.01; R=0.04, p=0.04; R=- 0.79, p=0.01). In the androgen metabolites, androstenedione, 5-AT, 11- keto Et, 11-betahydroxy Et, THS, and THE were significantly related to the residual volume in uroflowmetry (R=0.92, p=0.001; R=0.84, p=0.008; R=0.99, p=0.000; R=0.72, p=0.03; R=0.97, p=0.000; R=0.85, p=0.00). beta-THF/alpha-THF was significantly related to the maximum flow rate, the residual volume in uroflowmetry, the maximum urethral closure pressure and the functional urethral length (R=-0.76, p=0.02; R=0.67, p=0.04; R=0.74, p=0.02; R=-0.92, p=0.000). alpha cortol was significantly related to the residual volume in uroflowmetry, the maximum urethral closure pressure and the functional urethral length (R=0.81, p=0.01; R=0.71, p=0.03; R=-0.87, p=0.000). Of the estrogen metabolites, estrone (E1) was significantly related to the normal desire to void (R=0.68, p=0.04) and 17beta-estradiol/estrone was also significantly related to the normal and strong desire to void (R=-0.70, p=0.03 and R=-0.74, p=0.02, respectively). The urinary progesterone and androgen metabolite concentrations were positively related to the residual volume in uroflowmetry and positively or negatively related to MUCP and FUL. However, the urinary estrone concentration was positively related to the normal desire to void and 17beta-estradiol/estrone was significantly related to the normal and strong desire to void. PMID- 12728470 TI - Anti-Toxoplasma gondii antibodies in haemodialysis patients with chronic renal failure. AB - This study aimed to determine the prevalence of anti- Toxoplasma gondii antibodies in haemodialysis patients with chronic renal failure (CRF). METHODS: One hundred and seventy three haemodialysis patients, and 40 healthy controls, were studied for the prevalence of anti-Toxoplasma gondii antibodies by a micro enzyme-linked immunosorbent assay (ELISA). Anti-T. gondii IgG antibodies were detected in 97 (56.06%) haemodialysis patients and 8 (20%) controls with a statistical significance. In addition, anti-T. gondii IgM antibodies were detected in 1.73% of patients, but none of the controls. In conclusion, a high percentage of positivity for Toxoplasma antibodies in patients with CRF undergoing haemodialysis was noticed, thus parasitological surveys of CRF patients should be periodically performed to prevent the possible dissemination of toxoplasmosis through the dialysis procedure. PMID- 12728471 TI - Increased expression of intracellular HLA-DM but not on the surface of blood monocyte-derived dendritic cells during maturation. AB - Cutaneous dendritic cells (DCs), Langerhans cells (LCs) and dermal dendritic cells (DDCs), are present in an immature state. The maturation of DCs is crucial for initiating an immune response. Since HLA-DM has an important role for antigen presentation, an increase in HLA-DM expression according to the maturation of blood monocyte-derived dendritic cells (MoDCs), which have similar characteristics with DDCs, is expected. Therefore, the aim of this study was to determine whether or not HLA-DM expression in MoDCs is related to maturation at each culture day (from day 0 to day 13) by flow cytometry. This was compared with the functional changes related to the maturation of MoDCs. MoDCs were generated by culturing human peripheral blood monocytes in the presence of GM-CSF and IL-4 for 7 days, which were followed by subsequent treatment with a cytokine cocktail (GM-CSF, IL-4, IL-1beta, TNF-alpha, IL-6 and PGE2) for the maturation of MoDCs. The intracellular HLA-DM was expressed in the immature MoDC. A sudden 3 to 8 fold increase in the intracellular HLA-DM expression was observed after treatment with a cytokine cocktail. HLA-DM was weakly expressed on the surface of the immature MoDC, but it seemed to be decreased with maturation. This study indicated that the intracellular HLA-DM expression increased, but not on the MoDC surface during maturation. This was despite the fact that HLA-DM expression was noted not only on the surface but also in the intracellular in the MoDC. PMID- 12728472 TI - Cellular proliferative effect of dexamethasone in immortalized trabecular meshwork cell (TM5) line. AB - Dexamethasone (DEX), one of the corticosteroid hormones, is one of the most common therapeutic strategies in ophthalmological treatment. Despite its widespread use and clinical efficiency, little is known regarding the specific effects of DEX on cell growth, differentiation and cell death in human trabecular meshwork cells. The presence of the glucocorticoid receptor (GR, dexamethasone receptor) in TM-5 cell line, which was derived from the primary human trabecular meshwork cells, was verified by RT-PCR and western blot analysis. The effects of DEX on the cellular proliferation of TM5 cells were measured by a BrdU incorporation assay. Western blot analysis were used to examine the effects of DEX on the Ras/MEK/ERK signaling pathway. The total Ras, MEK1/2 and ERK1/2 protein levels as well as the levels of activated (phosphorylated) form were both significantly increased by the DEX treatment for 5 days. Both MEK1/2 and ERK1/2 were significantly activated by phosphorylation after 10 minutes. The dependence of this increased cell proliferation on GR activation by DEX and the sustained activation of ERK was examined using RU486 (a GR inhibitor) and U0126 (a MEK inhibitor). Both RU486 and U0126 prevented the induction of cell proliferation by the DEX treatment in the TM5 cells. In conclusion this study demonstrated that GR is expressed in TM5 cells. Secondly, DEX treatment for 5 days stimulates cell proliferation in TM5 cells, and that this increased proliferation effect is mediated by the Ras/MEK/ERK pathway. PMID- 12728473 TI - Investigation of colposcopy information on the internet. AB - The Internet is a massively expanding body of information which includes medical resources. It is easier than in the past for anyone with some knowledge and effort to access the Internet. Medical professionals as well as patients have a similar ability to research a medical topic through the Internet. As medical professionals, we are concerned whether the medical information found on the Internet by patients is current and accurate. There are relatively few reports that critically examine where patients and family members acquire medical information. To assess the accuracy and completeness of information regarding colposcopy on the Internet, we categorized and defined the information on the Internet through systematic study. We chose eight search engines available on the homepage of Explorer to search and analyze the information obtained from a search for the topic of colposcopy. In addition, we analyzed whether the colposcopy information contained peer-review or reference in order to evaluate its reliability. The numbers of uniform/universe resource locators (URLs) varied considerably, ranging from 11 to 23300, when we tested the search engines using different combinations of capitalization and spellings. Descriptions of information from 181 sites located by a search engine (Yahoo) were also evaluated into five types, ranging from diagnostic to anecdotal information. In terms of the peer-review system, inaccurate or misleading information was found on web pages which might lead patients to doubt and distrust their medical specialists whose information was contradictory to that which they had obtained themselves. We recognize that the Internet has a positive and potential power in partnership with medical professionals to educate patients, helping them to overcome their disease. Nevertheless, we suggest the need to take an active role in identifying the Web sites containing accurate medical information and evaluating their quality. PMID- 12728474 TI - Effect of bedding control on amount of house dust mite allergens, asthma symptoms, and peak expiratory flow rate. AB - This quasi-experimental study was designed to investigate the effect of bedding control on the amount of house dust mite (HDM) allergens, asthma symptoms, and peak expiratory flow rate (PEFR) in asthmatics sensitive to HDMs. The subjects in the study were drawn from patients receiving treatment at the allergy clinics of three university-affiliated hospitals in Seoul. Forty-two patients without prior practice of the bedding control used in this study were selected. They commonly showed bronchial asthma caused by HDMs, and exhibited strong positive points (more than 3 points) in skin prick test (D. farinae, D. pteronyssinus), and positive response in both fluoro-allergosorbent test (FAST), and PC20 methacholine test. Of the subjects, alternatively, 22 were assigned to the experimental group and 20 to control group. Bedding control consisted of the use of outer cotton covers, boiling them for 10 minutes fortnightly, and disinfecting bedding by sunlight fortnightly. The experimental group was under bedding control for 4 weeks. The data were collected from October 2000 to January 2001. The results were as follows: 1. After bedding control, the total amount of HDM allergens decreased significantly in the experimental group. However there was no significant difference in the decrease of the amount of HDM allergens between the two groups. 2. Of the asthma symptoms, there was significant difference only in the decrease of the frequency of dyspnea, and in the increase of sleeping disturbance between the two groups after bedding control. 3. After bedding control, PEFR increased in the experimental group whereas it decreased in the control group. However, neither change was significant. The above findings indicate that bedding control improved several asthma symptoms in asthmatics sensitive to HDMs. Accordingly, we suggest that bedding control is adopted as a useful nursing intervention in the field. PMID- 12728475 TI - Transverse myelitis in a patient with primary antiphospholipid syndrome. AB - The neurological manifestations of antiphospholipid syndrome (APS) are diverse. Transverse myelitis (TM) is an uncommon, but well-known neurological complication of systemic lupus erythematosus (SLE). On the other hand, the reported cases associated with primary APS are extremely rare. To our knowledge, this is the first report of TM in a patient with primary APS in Korea. A 32-year-old male patient was admitted with the sudden onset of numbness, a tingling sensation, and weakness in both lower extremities. He had a 19 months history of external iliac and femoral arterial thromboses prior to admission. The laboratory results indicated the presence of anticardiolipin antibodies of the IgG class and lupus anticoagulant. No other autoantibodies were detected and there were no apparent clinical manifestations of SLE or multiple sclerosis. A T2-weighted magnetic resonance (MR) image showed swelling and increased intensity of the cervical and thoracic spinal cord between C6 and T7 with slight enhancement by contrast medium. After steroid pulse therapy, the patient's symptoms were gradually relieved and the abnormal findings on MR imaging disappeared. PMID- 12728477 TI - A case of late infantile neuronal ceroid lipofuscinosis. AB - Neuronal ceroid lipofuscinosis, which is also known as Batten-Bielschowsky disease, is a group of neuro degenerative disorders, associated with various progressive symptoms including seizures, dementia, visual loss and cerebral atrophy. We experienced a case of late infantile neuronal ceroid lipofuscinosis in a 6-year-old boy who had progressive myoclonic seizures, ataxia, rapid psychomotor deterioration and visual loss. Photic stimulation at 2 to 5 Hz elicited a discrete spike and wave discharges in the occipital region on an electroencephalogram. Magnetic resonance imaging of the brain showed generalized cerebral and cerebellar atrophy. An electron microscopic examination of the skin revealed characteristic curvilinear inclusion bodies. An optic fundoscopy revealed a devastated retina and severe optic atrophy. We report this case with the brief review of related literature. PMID- 12728476 TI - Rhabdomyolysis in celiac disease. AB - A 12-year-old female presented with chronic diarrhea, fatigue, failure to thrive, sudden weakness of her upper and lower extremities and inability to walk. On neurological examination, atrophy was found of the lower extremity muscles, coupled with muscle weakness. Hypokalemia and a high creatine kinase (CK) level were detected. Antigliadin IgA, IgG and antiendomysial antibodies were positive. A duodenal biopsy revealed the classical findings of celiac disease. To our knowledge this is the first childhood case of celiac disease presenting with rhabdomyolysis. PMID- 12728478 TI - Homozygous exon 4 deletion in parkin gene in a Korean family with autosomal recessive early onset parkinsonism. AB - The gene responsible for autosomal recessive parkinsonism, parkin, has recently been identified on chromosome 6q. It has been shown to be mutated in Japanese and European families, most of whom had early-onset parkinsonism. Here, we present a family with young-onset parkinsonism of an autosomal recessive inheritance. A homozygous exon 4 deletion in the parkin gene was found in 3 family members. To the best of the authors' knowledge, this is the first report in Korea of familial parkinsonism with the parkin gene mutation. PMID- 12728479 TI - Spinal epidural metastasis of cerebral oligodendroglioma. AB - A 50-year-old male patient with right frontal oligodendroglioma underwent subtotal resection on three separate occasions and, 10 months later, exhibited right frontal oligodendroglioma and extracranial metastasis. Spinal magnetic resonance imaging (MRI) demonstrated the existence of an enhancing mass lesion with evidence of posterior epidural compression at the 10th-11th thoracic level, not involving the vertebrae. A bone scan of the spine appeared normal, but showed evidence of hot uptake in the pelvis and femur. This report concerns a patient who developed a fatal and clinically unexplained, pancytopenia 3 months after the removal of a spinal epidural oligodendroglioma. Oligodendroglioma with metastasis outside the central nervous system is extremely rare, and only a few cases have previously been reported. A brief review of the literature with an emphasis on the mechanisms of tumor cell dissemination is presented. PMID- 12728480 TI - A case of superficial epithelioma with sebaceous differentiation. AB - Superficial epithelioma with sebaceous differentiation (SESD) is a rare benign neoplasm with peculiar histopathologic characteristics, which occurs in aged skin. We report upon a case of SESD occurring on the left upper back, which histopathologically is a superficial, multilobular tumor with numerous basaloid cells mixed with sebaceous cells attached to the overlying epidermis. This case represents a solitary benign neoplasm without any associated malignancy. PMID- 12728481 TI - A case of thyroid storm due to thyrotoxicosis factitia. AB - We describe a case of thyroid storm due to thyrotoxicosis factitia, which was caused by the ingestion of excessive quantities of exogenous thyroid hormone for the purpose of reducing weight. An 18-year-old female was admitted to the hospital 24 hours after taking up to 50 tablets of synthyroid (1 tablet of synthyroid : levothyroxine 100 microg). Because of her stuporous mental state and acute respiratory failure, she was intubated and treated in the intensive care unit. After reviewing her history carefully and examining plasma thyroid hormone levels, we diagnosed this case as a thyroid storm due to thyrotoxicosis factitia. Her thyroid function test revealed that T3 was 305 ng/dL, T4 was 24.9 microg/dl, FT4 was 7.7 ng/dL, TSH was 0.05 micro IU/mL and TBG was 12.84 microg/mL (normal range: 11.3 - 28.9). TSH receptor antibody, antimicrosomal antibody, and antithyroglobulin antibody were negative. She was recovered by treatment, namely, steroid and propranolol, and was discharged 8 days after admission. Thyroid storm due to thyrotoxicosis factitia caused by the ingestion of excessive thyroid hormone is rarely reported worldwide. Therefore, we now report a case of thyroid storm that resulted from thyrotoxicosis factitia caused by the ingestion of a massive amount of thyroid hormone over a period of 6 months. PMID- 12728482 TI - Metastatic renal cell carcinoma of the gallbladder. AB - Metastatic renal cell carcinoma is renowned for its potency to spread to almost any organ of the body; however metastasis to the gall bladder is very rare. We present a case of a 48 year old man who initially demonstrated renal cell carcinoma, and in who gallbladder metastasis was later detected. A review of the literature revealed only a small number of cases of renal cell carcinoma metastasizing to the gallbladder, and these were primary found upon necropsy. Gallbladder metastasis in this case was detected clinically. PMID- 12728483 TI - Segmental jejunal lipomatosis--a rare cause of intestinal obstruction. AB - A rare case of a segmental small intestinal (jejunal) lipomatosis is described. A 33-year-old male was admitted with a clinical diagnosis of an acute intestinal obstruction. A plain erect abdominal x-ray showed multiple air-fluid levels. On an exploratory laparotomy, a jejunojejunal intussusception was found secondary to a segmental submucosal lipomatosis. This was treated by a segmental resection and anastomosis, which resulted in a complete cure. Here we present this case with a review of the relevant literature. PMID- 12728484 TI - [Changes in whole blood ionized magnesium concentration during cardiac surgery employing St. Thomas' cardioplegic solution]. AB - BACKGROUND: Although there is growing evidence to suggest that magnesium supplementation to patients undergoing cardiac surgery is beneficial, the way to administer magnesium is not established. Moreover in Japan St Thomas' cardioplegic solution, containing a high level of magnesium is widely used and the effect of such magnesium-rich cardioplegic solutions on blood magnesium concentration has not been well defined. METHODS: We measured ionized magnesium concentrations (iMg) during cardiac surgery employing St Thomas' solution. Patients were divided into four groups. Group 1 patients were adults and group 2 were children, both of whom received St. Thomas' solution. Group 3 patients underwent cardiopulmonary bypass but did not receive any cardioplegic solution. Group 4 patients underwent off-pump coronary artery bypass grafting. RESULTS: In cardioplegia group (group 1 and 2) iMg was higher than the normal reference range at periods of rewarming, immediately postbypass, and at the end of the operation. iMg at immediately postbypass was related to the total amount of cardioplegic solution. In non-cardioplegia group (group 3 and 4) progressive decrease of iMg was observed throughout the operation. CONCLUSION: Because magnesium in cardioplegic solutions has substantial effect on perioperative iMg, it is crucial to measure iMg to avoid overdose of magnesium when magnesium-rich cardioplegic solutions are employed. PMID- 12728485 TI - [The effects of inhaled carbon monoxide on lung injury in rats caused by lipopolysaccharide]. AB - BACKGROUND: Carbon monoxide (CO) has long been considered a toxic substance. Recent studies have revealed that CO may play an important role in intercellular signaling. We hypothesized that CO modulates the inflammatory mechanisms. METHODS: SD rats (each study group consisting of 7 animals) inhaled 250 ppm of CO one hour prior to LPS challenges. These animals were incubated for a particular period of time (four different length of time; 1, 2, 4, and 8 hours). The control group (each study group consisting of 6 animals) had been left in the room air. Both groups were instilled with LPS (1 mg) into the lungs. At the end of each period, animals were exsanguinated, broncho-alveolar lavage (BAL) and blood sampling were performed, and a part of the small intestine was harvested. PMN numbers, protein, TNF, and IL-10 concentrations in BAL fluid were measured. Plasma TNF and IL-10 were also measured. The Wet/Dry ratio of a small intestine was calculated. RESULTS: In the CO-inhalation group, the anti-inflammatory cytokine IL-10 concentration in the BAL fluid was higher at 8 hours after LPS challenge than the counterpart of room air group (37.6 +/- 11.4 pg.ml-1 vs. 92.8 +/- 31.5 pg.ml-1, P < 0.05). W/D ratio decreased (3.29 +/- 0.2 vs. 2.94 +/- 0.06, P < 0.05). CONCLUSION: These findings indicated that CO inhalation might modulate the inflammatory response to LPS. Further study is needed to prove the therapeutic role of CO inhalation without serious adverse effects. PMID- 12728486 TI - [Dynamic cerebral blood flow autoregulation during sevoflurane anesthesia and TIVA]. AB - BACKGROUND: Dynamic cerebral blood flow autoregulation during sevoflurane anesthesia and total intravenous anesthesia (TIVA) is unclear. We examined the cerebral circulation autoregulation during anesthesia by sevoflurane or TIVA. METHODS: We measured mean blood pressure (MBP) and blood flow velocity of the middle cerebral artery by a transcranial Doppler ultrasonography before and during anesthesia using sevoflurane (volatile induction and maintenance of anesthesia (VIMA) group) and using propofol and fentanyl (TIVA group), and the relationship between changes in MBP and cerebral blood flow velocity was evaluated using the method of transfer function analysis. We calculated transfer gain and coherence by cross-spectrum from autospectra of MBP and cerebral blood flow velocity. RESULTS: Transfer gain during anesthesia by TIVA in the low frequency range and high frequency range was near 1 cm.sec-1.mmHg-1. It was about equal to the value of transfer gain before anesthesia. But transfer gain during anesthesia by VIMA was above 2 cm.sec-1.mmHg-1. CONCLUSION: These results suggest that TIVA by propofol and fentanyl maintains the dynamic autoregulation of cerebral blood flow, but sevoflurane impairs the autoregulation. PMID- 12728487 TI - [The effect of intrathecal fentanyl added to hyperbaric bupivacaine for caesarean section]. AB - BACKGROUND: Management of cesarean section with spinal anesthesia is often accompanied with intraoperative nausea and pain. In a randomized controlled study, we explored the effect of intrathecal fentanyl on the characteristics of subarachnoid block in patients undergoing cesarean section. METHODS: Twenty-four healthy parturients scheduled for elective Cesarean section were allocated to receive either fentanyl 0.3 ml (15 micrograms) or 0.9% saline 0.3 ml added to 0.5% hyperbaric bupivacaine 2.0 ml given intrathecally in the right decubitus position (n = 12 in each group). Level of sensory blockade was evaluated with cold test and intraoperative use of antiemetics and analgesics was recorded. RESULTS: The maximum level of sensory blockade was significantly higher in the fentanyl group as compared with the control group (P = 0.019). Use of intraoperative antiemetics was significantly less often in the fentanyl group (P = 0.007). The required amount of intraoperative analgesics was smaller in the fentanyl group, although the difference was not significant (P = 0.11). No remarkable side effects, such as respiratory depression and hypoxia were observed. Apgar scores in the newborn were similar. CONCLUSION: Addition of intrathecal fentanyl to hyperbaric bupivacaine in parturients undergoing cesarean section improved quality of anesthesia without producing significant side effects. PMID- 12728488 TI - [Postoperative continuous epidural analgesia with portable disposable infuser pump: review of 1500 cases]. AB - BACKGROUND: We have been offering active postoperative analgesia service (PAS). In order to further enhance the quality of PAS, we reviewed 1500 cases of postoperative continuous epidural analgesia (PCEA). METHODS: Postoperative patients received epidural administration of a mixed analgesic solution employing bupivacaine, morphine, or fentanyl using a portable disposable infuser pump (PDIP) for at least 72 hours. Analgesic effects were assessed by using Prince Henry's scoring and the demand for additional analgesics. The frequency and causes of discontinued infusion were also evaluated. RESULTS: 83.9% of cases showed no complications. However, in 4.7% of cases infusion was discontinued because of side effects (nausea, vomiting, pruritus, lower limb motor block, and hypotension, etc.), in 4.1% because of dislodgment of the epidural catheter, in 3.4% because of disconnection, in 2.0% because the patient removing the catheter, and in 0.8% because of the condition of the catheter insertion site. CONCLUSION: These results indicate that to improve our method of PCEA with PDIP, we must re assess our regimen used for continuous epidural infusion for postoperative pain relief, and develop measures to prevent side effects and complications. PMID- 12728489 TI - [Bispectral index (BIS) in infants anesthetized with sevoflurane in nitrous oxide and oxygen]. AB - BACKGROUND: The bispectral index (BIS) has been shown to be useful in monitoring a degree of hypnosis in anesthetized adults. Although several studies have been performed to evaluate BIS in pediatric patients, it is unclear whether BIS monitor can be applied to infants. This study was designed to evaluate if the BIS monitor can be used in infants as a measure to monitor a degree of hypnosis. METHODS: Forty-three infants were divided into two age groups according to their age: group A (3-6 months; n = 31) and B (7-12 months; n = 12). And the patients in the younger group were randomly allocated to one of two groups, A (+) (premedicated with oral midazolam 0.5 mg.kg-1; n = 19) and A (-) (without premedication; n = 12). BIS values and other parameters were recorded at a steady state of end-tidal sevoflurane concentration (2.5, 2, and 1.5%) and immediately before extubation. RESULTS: At each concentration of sevoflurane and extubation, the BIS values were lower in group A (+) than in group B (P < 0.05). In infants < or = 6 months of age, premedication did not affect the BIS values. CONCLUSION: There were significant differences of BIS values between < or = 6 months and > or = 7 months old infants. BIS values should be interpreted cautiously in infants younger than 6 months. PMID- 12728490 TI - [Occupational exposure of operating room staff to anesthetic gases during inhaled induction--a comparison with intravenous anesthesia induction]. AB - BACKGROUND: The risk of occupational exposure to waste anesthetic gases still remains during inhaled induction. In this study we investigated how much we were occupationally exposed to anesthetic gases during induction period. METHODS: Twenty-six adult patients were induced with sevoflurane 5% using a face mask for three minutes and maintained with sevoflurane 1% after end-tracheal intubations (IH-Group). Twenty-two adult patients were induced with intravenous anesthetics and maintained with sevoflurane 1% after end-tracheal intubations(IV-Group). The concentration of sevoflurane was measured by Multi-gas Monitor 1302 (Bruel & Kjaer: Denmark) every 70 seconds. Sample gas was suctioned from breathing zone of anesthesiologists. All of our operating rooms are equipped with waste gas scavenging system. RESULTS: The peak concentration of sevoflurane is significantly higher in IH-Group (15.91 +/- 22.64 ppm) compared with IV-group (0.36 +/- 0.25 ppm). The period when sevoflurane concentration exceeded 0.5 ppm is significantly longer IH-Group (18.55 +/- 10.51 min.) compared to IV-Group (1.92 +/- 4.56 min.). CONCLUSION: The induction with intravenous anesthetics is a better method in order to reduce occupational exposure of anesthesiologists to anesthetic gases. PMID- 12728491 TI - Aspirin reversal--an incremental administration of aspirin shortens the bleeding time prolonged by low-dose aspirin. AB - In summary, an incremental dosage of aspirin (660 mg) shortens the bleeding time prolonged by daily low-dosage aspirin (81 mg) in healthy subjects. The "Aspirin Reversal" phenomenon might be useful to control excessive bleeding during surgery in patients taking daily low-dosage aspirin as anti-coagulants. PMID- 12728492 TI - [Sudden onset of EDTA-dependent pseudothrombocytopenia in a patient scheduled for open heart surgery]. AB - A 60-year-old woman scheduled for mitral and aortic valve replacement had sudden onset of thrombocytopenia without clinical symptoms. The platelet count was found to decrease after the sampling. Microscopic examinations confirmed platelet aggregations. Changing anticoagulant added to blood samples from EDTA to heparin resolved such platelet aggregations. This phenomenon was diagnosed as demonstrating EDTA-dependent pseudothrombocytopenia and the operation was performed as scheduled without platelet transfusion. Postoperative course was almost uneventful and the patient was discharged on 26th day after surgery. EDTA dependent pseudothrombocytopenia must be ruled out when patients have thrombocytopenia without certain causes such as infections, drugs, or autoimmune diseases. PMID- 12728493 TI - [A case of post-radiation therapy patient with difficulty in intubation unexpected preoperatively]. AB - We experienced a post-radiation therapy patient who had a narrow trachea, and presented with unexpected difficulty for intubation. His trachea was narrowest at 2 cm below the glottis. The fiberscope barely could pass the narrowest part of the trachea. We speculate that the radiation therapy induced his tracheal constriction. Careful attention is necessary when examining the post-radiation therapy patients. Multiple cervical radiographs are necessary in such a case. PMID- 12728494 TI - [Perioperative management of a patient with factor V Leiden mutation]. AB - Although the factor V Leiden mutation had not been reported in Japan, we experienced the perioperative management of a patient with factor V Leiden mutation. A 38-year-old Caucasian woman with factor V Leiden heterozygote underwent right lobectomy for thyroid tumor under general anesthesia. The postoperative course was uneventful. Heterozygosity increases risk of venous thrombosis sevenfold. A homozygote has a 20-fold increased risk of thrombosis. Heterozygosity coupled with ingestion of oral contraceptives or pregnancy increases the risk 15-fold. Anticoagulant therapy including the use of low molecular weight heparin is considered to be appropriate for perioperative management of the patient with factor V Leiden mutation. PMID- 12728495 TI - [Citizen's understanding of anesthesia from a questionnaire survey]. AB - BACKGROUND: In order to know general understanding or impression of anesthesia by citizens, we performed a questionnaire survey. MATERIALS AND METHODS: The participants for the survey were 218 citizens including 118 healthy persons with previous attendance to Kurume University Public Lecture for Citizens (group A) and 100 patients hospitalized in Kurume University Medical Center (group B). Their answers were collected prior to the lecture planned on the 39th Annual Meeting of Kyushu Society of Anesthesiologists. RESULTS: The recovery of the survey was 78.9%. The percentages of persons who wanted to know "How and by whom I am anesthetized" were 93.2% and 72.6% in groups A on B, respectively. The percentage of persons who wanted to know the name of surgeon was 91.7%. The answers to the question; "Which doctor do you think is responsible for the treatment of an intraoperative adverse event such as acute myocardial infarction?"; were in the order of an internist 37.5%, a surgeon 29.7% and an anesthesiologist 18.0%. CONCLUSION: We have realized through the questionnaire survey that the majority of citizens still consider an anesthesiologist as a technician merely putting patients into sleep for surgery. At the preoperative examination or round, we have to explain procedures and potential risks related to anesthesia to a patient and also have to answer patient's questions fully. We anesthesiologists should do more efforts to enlighten citizens at every opportunity so that citizens would understand our work fields and evaluate anesthesiologists correctly. PMID- 12728496 TI - [A new heat and moisture exchanger, Trach-Vent Plus, for patients with spontaneous respiration]. AB - A new heat and moisture exchanger, Trach-Vent Plus is now available in Japan. A heat and moisture exchanger, Trach-Vent and oxygen-supplying tool, Oxy-Vent are made in one unit as a Trach-Vent Plus. This is smaller and lighter than the combination of Trach-Vent and Oxy-Vent. Oxygen supplying tube can be connected to the side of the filter. There is a silicon membrane with slits in the center of the tool, to which patients's airway opens directly. The membrane prevents patients from dust, but suction of sputa is easy through the slits. This new heat and moisture exchanger, Trach-Vent Plus is useful to administer oxygen to intubated patients breathing spontaneously, especially with tracheostomy. PMID- 12728497 TI - [Reduction in the use of pulmonary artery catheter for cardiovascular surgery]. AB - In this study we have surveyed the rate of use of pulmonary artery catheter (PAC) in 319 cardiovascular operations from 1997 to 2001. The rate was decreasing year by year from 100%(6/6) in 1997, 77%(39/51) in '98, 32%(24/75) in '99, 20%(19/93) in 2000, to 8.5%(8/94) in 2001. Especially in elective coronary artery bypass grafting (CABG), an extreme reduction was found: 100%(3/3), 95% (19/20), 17%(6/35), 2.2%(1/45), 0%(0/53), respectively. There was no statistical significance between the use and hospital mortality. We consider that this reduction is due to improvement of the surgical technique and anesthetic management. As transesophageal echocardiography (TEE) has improved anesthetic management of the cardiac patients, we have employed it for all cases. We are sure that TEE contributed to the reduction in the catheter use. Cardiac functions monitored with PAC can be largely substituted with TEE except mixed venous oxygen saturation, but it is not continuous monitoring. Current indications of PAC is for patients with pulmonary hypertension and low cardiac output, as well as those who are predicted to be difficult for postoperative management, and those who have off pump CABG with inverted heart position, at our institution. Routine use of PAC for cardiac surgery should be reconsidered. PMID- 12728498 TI - [Usefulness of a new nasal cannula with an end-tidal carbon dioxide measuring port]. AB - BACKGROUND: During light sedation under local anesthesia, usually only percutaneous arterial oxygen saturation (SpO2) has been monitored for respiration. Measuring end-tidal carbon dioxide tension (EtCO2) has been difficult in such cases because of many air leaks from mask or cannula. We investigated the usefulness of a new nasal cannula with which EtCO2 can be monitored simultaneously with oxygen delivery. METHODS: Forty patients receiving epidural and spinal anesthesia were sedated with intravenous midazolam followed by propofol infusion. Oxygen 2l.min-1 was administered by a new nasal cannula of which one side of the face piece provides oxygen and the other side of it is an EtCO2 measuring port. SpO2 and EtCO2 were measured. RESULTS: The lowest SpO2 was 96 +/- 2%. The EtCO2 wave and value were demonstrated all the time during surgery in 24 patients, while in the other 16 patients sometimes it failed to indicate wave and value of EtCO2 when patients were snoring or moved their face. CONCLUSION: A new cannula with EtCO2 port is useful to oxygenate and measure EtCO2 simultaneously except when patients snore or move. PMID- 12728499 TI - [Investigation on nurses' knowledge of and experience in cardiopulmonary resuscitation and on nurses' knowledge of the guidelines for cardiopulmonary resuscitation and emergency cardiovascular care established in 2000--results of a survey at Asahikawa Medical College Hospital (second report)]. AB - BACKGROUND: We reported that the practical knowledge and skill on basic life support were not enough among nursing staffs. Therefore, to clarify the existing conditions and issues in CPR education we carried out a survey on knowledge of and experience in cardiopulmonary resuscitation (CPR) and on knowledge of the Guidelines for Cardiopulmonary Resuscitation and Emergency Cardiovascular Care (ECC) established in 2000. METHODS: Three hundred and four nurses at Asahikawa Medical College Hospital were asked to fill in questionnaires. RESULTS: The results show that more than 80% of the nurses are much interested in CPR. Most of the nurses had received education and training in CPR as students or after graduation. However, cases of cardiopulmonary arrest and CPR were experienced by only about 40% of the nurses. Most of the nurses had never heard of the Guidelines for CPR and ECC in 2000. CONCLUSION: The results of this survey demonstrate the need to provide more education (on CPR) to nursing staff. PMID- 12728500 TI - [The current anesthetic practice in the United Kingdom]. AB - In the current anesthetic practice in the United Kingdom (UK), there are some unique characteristics that are not necessarily familiar to the Japanese anesthesiologists. The author has obtained first-hand experience in anesthesia in the UK as a qualified anesthetist through the fellowship program between the Royal College of Anaesthetists and the Japanese Society of Anesthesiologists. The major differences in the UK anesthetic practice from its Japanese counterpart include dissimilarity in the use of drugs, the existence of "anaesthetic rooms", the expertise of perfusionists and operating department practitioners, and excellent systematic support from hospital professionals such as statisticians and graphics experts. Although the UK anesthetic practice may not be perfect, it certainly provides practical wisdom that may be useful for Japanese anesthesiologists. PMID- 12728501 TI - [Cerebral ischemia and free radical]. PMID- 12728502 TI - [Brain circuits involving emotional memory of stress: seeking the biological background of PTSD]. PMID- 12728503 TI - [Cerebral lateralization in spoken language processing measured by multi-channel near-infrared spectroscopy (NIRS)]. AB - Language processing is lateralized to the left hemisphere in 95% of people as assessed by invasive techniques like Wada test(intracarotid amobarbital test) and cortical electric stimulation. Here we report a new non-invasive method of assessing language dominance by measuring speech-evoked local cerebral responses in hemoglobin(Hb) concentration with near infrared spectroscopy(NIRS). The activation of the bilateral auditory areas to phoneme and pitch contrasts was investigated in twenty right-handed(RH) and ten non-right-handed (NRH) normal hearing native speakers of Japanese with a 24-channel NIRS system using 830 and 780 nm lasers. Three analysis-synthesized, inflected forms of a Japanese verb/iku/; (a)/itta/, (b)/itte/, and (c)/itta?/, were used as stimuli, delivered diotically at a rate of one word per second on average. In phoneme and pitch contrast conditions, a baseline block containing (a) and a test block including (a) and (b) (phoneme-contrast) or (a) and (c) (pitch contrast) were alternated and repeated several times. Laterality indices, LI = (L - R)/(L + R), calculated from the peaks of the selectively averaged Hb responses in the left(L) and right(R) auditory areas, were compared between the two contrast conditions. A significant leftward shift of LI in the phoneme condition relative to the pitch condition was observed in 85% of the RH subjects, with a significant group difference in the same direction. In contrast, 50% of the NRH subjects showed an inverse shift of LI between the two conditions, but no significant overall shift of LI was observed in this group. These results suggest that multi-channel NIRS could help non-invasively determine the dominant temporal cortex in speech perception, independent of speech production. PMID- 12728504 TI - [Therapeutic results of eight patients with intracranial ependymomas]. AB - Between 1985 and 2001, eight patients with intracranial ependymomas underwent surgery at our hospital. The cases included six infratentorial ependymomas, one supratentorial ependymoma and one supratentorial anaplastic ependymoma. Infratentorial ependymomas were classified according to origin and extension. The lateral type tumors originated from the lateral part of the fourth ventricle in four cases. The midfloor type tumors originated from the inferior half of the fourth ventricular floor in two cases. The three totally resected tumors were the lateral type tumors. The remaining one case with the lateral type tumor underwent nearly total resection of the tumor, since the tumor involved lower cranial nerves. All patients with the midfloor type tumors underwent incomplete resections of the tumors, because the tumors infiltrated into brain stem. Lower cranial nerve involvement and brain stem invasion implied incomplete resection and had the poor prognosis. In intracranial ependymomas, all four patients with total resections have been alive, whereas three of four patients with incomplete resections have died. The mean survival time of all patients with intracranial ependymomas was 127 months from the time of the initial surgery. There were no deaths in the patients with tumors showing MIB-1 index < 10% (n = 4). The mean survival time of the patients with tumors showing MIB-1 index > or = 10% (n = 4), was 30 months. The extent of the resection, the age of the patients and MIB-1 index are important factors in the outcome in patients with intracranial ependymomas. Two representative children aged less than 3 years with the midfloor type tumors were presented. In a patient treated with conventional radiation and chemotherapy, residual tumor repeatedly enlarged within 12 months despite several resections of the tumor. The patient died 32 months after the initial resection. In contrast, the other patient received multidisciplinary treatment including Linac stereotactic radiotherapy (SRT) with a marginal dose of 27 Gy in 9 fractions, have been still alive for 45 months after the initial resection. The residual tumor slightly decreased in size and remained stable without evident growth 12 months after SRT. SRT may provide good local control for patients with intracranial ependymomas and have a favorable impact on survival. PMID- 12728505 TI - [A case of hypoxic brain damage induced by severe asthma successfully treated by hypothermia therapy]. AB - We experienced a case of hypoxic brain damage induced by severe asthma who was successfully treated by hypothermia. A 20-year-old woman with a history of bronchial asthma suffered from severe respiratory distress and she stopped breathing for about 20 minutes. She was admitted to our hospital with respiratory arrest, deep coma, mydriasis and weak motor response to pain. She was intubated and mechanically ventilated with 100% oxygen. She was cooled down to 33 degrees C within 4 hours of her arrival. Her body temperature was maintained at about 33 degrees C for 2 days, and then gradually rewarmed. During hypothermia, PaCO2 was quite high(80-100 mmHg), but the intracranial pressure was kept low. After hypothermia therapy, she became free from consciousness disturbance and there were no neurological disorders except for mild myoclonus. Hypothermia has a possibility of effective therapy for patients with hypoxic brain damage after respiratory distress. PMID- 12728507 TI - [A case report of Lhermitte-Duclos disease with systematic AVMs]. AB - We report a case of Lhermitte-Duclos disease (LDD) with huge AVMs of left extremities. The patient is a 46-year-old woman, who was identified heart failure due to AVMs at 13 years old and underwent amputation of left arm and several embolizations for AVM of left leg. Following a loss of consciousness, she was admitted to our hospital at 46 years old. CT scan showed a low-density area in the vermis of cerebellum. MR imaging showed a tumor with characteristic parallel linear striation. She was treated with partial resection of tumor. Pathological findings were dysplastic gangliocytoma. She was diagnosed LDD by MR imaging and pathological findings. Some reports describe an association between LDD and Cowden disease. On the other hand, there are reports of an association between Cowden disease and AVM. However, to our knowledge, there is no report of an association between LDD and AVM. Although this case didn't be revealed an association with Cowden disease, we believe that this case is a very interesting one henceforth suggesting the association between LDD, Cowden disease, and AVM. PMID- 12728506 TI - [Combined imaging of epileptic foci and brain metabolism using MEG and FDG-PET]. AB - A 17-year-old woman developed left hemiparesis at the age 6 months. She had suffered from focal motor seizures associated with tonic extension of her left extremities since the age of 10 years. The interictal scalp EEG demonstrated frequent spike-and-slow-wave complexes dominantly in the right frontal area. MRI showed an old cerebral infarction in the right frontal lobe. Simultaneous recordings of magnetoencephalography (MEG) and EEG were obtained by using a 204 channel whole-head MEG system. Equivalent current dipoles (ECDs) calculated from epileptic spikes on MEG were scattered in the cortex adjacent to the lesion in the right frontal lobe. Positron emission tomography with 18-fluoro-2 deoxyglucose (FDG-PET) in the interictal state showed hypometabolism in the lesion and its adjacent area. The super-imposed images of the dipole and PET showed that epileptic foci surrounded the lesion. The multimodality imaging is useful for evaluation of patients with epilepsy for possible indication of surgery. PMID- 12728508 TI - [Back to the high school days: an abnormal remembering characterized by concurrent as well as alternating feeling to be in the past and present]. AB - After the successful treatment of a hypothalamic germinoma, a 31-year-old right handed male developed a difficulty in memory and admitted to our department for detailed evaluation. Neuropsychologically he showed no personality change, confabulation, misidentification, delusion or disorientation to physical time. Neither aphasia, dementia or frontal lobe dysfunction was found. However, he showed a moderate degree of anterograde amnesia and a retrograde amnesia for the last 5 years judged by a public events test. Moreover, his temporal markings of a correctly remembered event in the post high school days shifted strongly toward the high school days. This temporal location abnormality was largely limited to the events from the days of his graduation from the high school to the period 5 years prior to the present incident. He felt personal events happened during this period as if they had occurred in the high school days, although he well knew that they actually happened much later. At the same time he felt that memories of the high school days were abnormally vivid and recent. Most interestingly, he was found to have been experiencing a funny feeling that he was living in the high school days and the present at the same time. The feeling occurred either simultaneously or alternatively. In the latter instance, he would suddenly start behaving as if he were a high school student. He would start urging his wife to go to the school together immediately. This behavior would be over in several minutes. His consciousness was clear in these instances and there was no amnesia for these periods. A brain MRI with gadolinium enhancement showed a lesion extending in the hypothalamus, anterior thalamus, basal forebrain and midbrain bilaterally. After eight months this abnormality of temporal sensation disappeared. However, tendency to make the date of public event nearer to the high school period persisted. We hypothesize that impaired temporal estimation for an event recalled from the retrograde memory store and mis-arousal of familiarity evoked at the time of spontaneous recollection are responsible for double feeling of time of an experience. These phenomenon might be attributable to the partial destruction of the Papez' and the basolateral limbic circuits. PMID- 12728509 TI - [A case of partial fascicular oculomotor paresis caused by midbrain infarction]. AB - We report a 74-year-old man with an ischemic lesion in the ventral midbrain. He presented with contralateral ptosis and marked upward gaze paresis of the right eye. Neurological examination revealed partial oculomotor nerve palsy caused by impairment of the right levator palpebrae, superior rectus and inferior oblique muscles. This finding is highly suggestive of a possible lesion in the midbrain affecting the oculomotor fascicular fibers. Magnetic resonance images showed an ischemic lesion in the paramedian area of the right midbrain tegmentum. The coronal view of T 2-weighted imaging clearly demonstrated to be the site of lesions below the red nucleus. It seemed to be coincidental with the impaired site of involving the caudal part of oculomotor fascicular fibers emerging from the nucleus. This report is considered to be a typical case of partial fascicular oculomotor paresis based on impairment of the caudal part of oculomotor fascicles for the levator palpebrae, superior rectus, and inferior oblique muscles. This is a valuable case to be documented in which neurological site of lesions are consistent with those found in radiological study. PMID- 12728510 TI - [A survival case of fluminant meningitis and multiple organ failure as overwhelming postsplenectomy infection (OPSI) syndrome]. AB - A 39-year-old man, who had splenectomy following abdominal trauma 30 years previously, admitted to our hospital with high fever, headache and convulsion on January 8, 2002. Examination of cerebrospinal fluid (CSF) showed pleocytosis up to 10,000 cells/mm3 with polymorphonuclear cells dominant and elevated protein to 330 mg/dl and reduction of glucose to 15 mg/dl. Streptococcus pneumoniae was detected at the culture of CSF and blood. We diagnosed him as having pneumococcal meningitis as overwhelming postsplenectomy infection(OPSI) syndrome. After administration of carbapenem antibiotics, methylprednisolone (1,000 mg) and immunoglobulin, he survived without any complications. Splenectomized patients are likely to suffer from severe infections, such as sepsis and meningitis, which is called OPSI syndrome. The course is rapid, the clinical symptoms are serious, and the prognosis is very poor. It is important for splenectomized patients to receive the vaccine, antibiotic prophylaxis and seek medical attention at the earliest sign of minor infection. PMID- 12728511 TI - [A rare massive parenchymal metastasis in central nervous system from abdominal non-Hodgkin lymphoma: effectiveness of stereotactic radiosurgey: case report]. AB - A 62-year-old female presented with a rare massive parenchymatous metastasis from abdominal malignant lymphoma. Computed tomography and magnetic resonance image revealed a large enhanced mass in the right basal ganglia. 123I-IMP SPECT showed increased uptake on both early and delayed images. A stereotactic biopsy was performed; histological examination revealed a diffuse large B-cell malignant lymphoma. The patient underwent stereotactic radiosurgery (SRS). Short-term cliniconeuroradiological follow-up showed both neurologic improvement and virtually complete disappearance of the tumor. Our findings suggest that 123I-IMP SPECT can help differentiate malignant lymphoma from benign lesions and other malignant brain tumors. In addition, SRS with conventional radiotherapy may be an effective therapeutic strategy to control malignant lymphoma. PMID- 12728512 TI - [A case of neurofibromatosis 2 starting from left oculomotor palsy]. PMID- 12728513 TI - Phenomenology of genius and psychopathology. AB - The relationship between genius and madness has been a subject of interest since the beginning of critical and philosophical thinking. Thus, Aristotle, in the Book XXX of the Problemata, asks himself "why are all extraordinary men in the fields of philosophy, politics, poetry and art melancholic?, adding afterwards: "...and some of them in such a way that they may suffer from pathologic manifestations whose origin is in the black bile". In the past decades the German author Tellenbach studied the personalities of several geniuses, both from fiction, such as Hamlet, and from reality, such as the writer von Kleist, concluding that they suffered from a specific form of depression that he called "Schwermut" (melancholy), which was supposedly different from the narrowly defined illness of depression. Other work done on this subject is the extensive study by the North American author Kay Jamison, who, after researching the biography and the tree of a long list of writers, composers and musicians, concluded that all of them had suffered to some degree from a bipolar disorder. This author strives to carry out a phenomenology of genius, and he finds that, together with other essential features, the geniuses always show forms of experiencing and/or of behaving which do not fall within the range that is considered normal, although they can not always be classified as pathological. His study is based on the analysis of the life and the work of three men whose genius could not be doubted: the naturalist Alexander von Humboldt, the philosopher Soeren Kierkegaard and the poet Rainer Maria Rilke. This author specially focuses on the last named, since in his later work he explicitly meditated on the suffering that has meant for him his condition of genius and what he considered the only way to overcome them: to be faithful to the work of art, whose fulfilment was imposed on him--to a certain degree from the endogenous -as an unavoidable imperative. PMID- 12728514 TI - Interdisciplinary collaboration: the obvious need and basic structures. AB - In the last decade there has been an exponential increase in knowledge in the field of medicine in general and particularly in Neurosciences. In order for the continuation of progress, it is important for professionals in all fields to collaborate with each other. Interdisciplinary collaboration is an absolute must to be able to obtain new knowledge, quickly and efficiently. Another important aspect is for professionals to collaborate with industry--especially the pharmaceutical industry. The global pharmaceutical companies account for a large amount of resources including personnel, knowledge, and especially funds. Although interdisciplinary collaboration is essential, many times potential partners overlook the obvious need for their own personal ongoing politics. Such competition needs to be overcome and new fields with multiple perspectives from different specialties will lead the way through the new millennium. PMID- 12728515 TI - Current status of general hospital psychiatry in Japan. AB - The current status of general hospital psychiatry was overviewed to clarify the progress and the stagnation. To facilitate well-organized community psychiatric care, general hospital psychiatry should play a central role in psychiatry. The policy makers of the Japanese mental health system should place a special emphasis on general hospital psychiatric beds to further process of shifting from a hospital based to a community based psychiatry system. It is also necessary that general hospital psychiatry should become more aggressively involved in community psychiatry, e.g. emergency psychiatry. Consultation-liaison (C-L) psychiatry has been quickly developed and become one of the main psychiatric fields. For further development, a multidisciplinary team approach with co medical staffs is necessary to supply efficient and effective care to medically ill patients. A proactive model of C-L care rather than a doctors' needs model should also be considered. Well designed research evaluating the efficiency and effectiveness of C-L activities in medical settings needs to be done to increase funding to general hospital psychiatry. This research evidence would also lead to a more fully integrated general hospital psychiatry into the practice of medicine and catch up with the ongoing medical reform in Japan. PMID- 12728516 TI - Consultation-liaison psychiatry in Korea. AB - The Korean Psychosomatic Society, our national organization for consultation liaison (C-L) psychiatry was formed within the Korean Neuropsychiatric Association (KNPA) in 1992. Since then, there has been increasing interest in C-L activities in general hospitals. All psychiatry departments in university teaching hospitals offer C-L experiences as part of the rotation for residents. Recently, there have been increasing research activities in C-L psychiatry. However, there are some current obstacles to further development of C-L psychiatry. No reimbursement for C-L activities is one of the most pressing problems facing C-L psychiatrists. Insufficient staffing at C-L services and stigmatization of mental illness are also problems to be dealt with. In order to resolve those issues, a more organized approach to demonstrate the usefulness of psychiatric C-L activities will be needed. PMID- 12728517 TI - Consultation-liaison psychiatry: an international perspective. AB - Consultation-liaison (C-L) psychiatry is the specialty of physical/psychiatric comorbidity and somatization. As such, it addresses the commonest forms of psychiatric presentation in the community; it is mainstream psychiatry. Its name reflects the fact that most of the patients seen by C-L psychiatrists have to be managed jointly with a non-psychiatrist doctor and other professionals. Because of this, the degree of development of the specialty and its mode of operation differ from country to country, depending on the organization of health services and the varying attitudes towards the disorders in which C-L psychiatry specializes. These differences permit a refreshing opportunity to re-examine the ways in which patients with physical/psychiatric comorbidity and somatization can be helped. The International Organization for Consultation-Liaison Psychiatry was formed to facilitate this process. It has identified challenges that are universal, and which if not met rapidly and efficiently will damage our ability to provide appropriate care for the patients whom we serve. PMID- 12728518 TI - Introduction of a board certification system for the Japanese Society of General Hospital Psychiatry. AB - The importance of board certification systems is increasing in parallel with changes in the social conditions surrounding general medical practice. The same is also the case for consultation-liaison psychiatry, a subspecialty of psychiatry. As a consequence, in April 2001 a board certification system for the Japanese Society of General Hospital Psychiatry was introduced. The clinical abilities required for liaison psychiatrists in this system can be summarized as follows: the ability to adequately treat patients with physical/psychiatric comorbidity or somatization, the ability to form an appropriate and adequate relationship with patients with physical diseases and to collaborate with medical and surgical professionals, and to have a good social and ethical awareness of general medical practice. The conditions required to obtain certification and the issues that remain to be addressed are also discussed. PMID- 12728519 TI - The journey to subspecialization in psychosomatic medicine (or consultation liaison psychiatry): a United States experience. PMID- 12728520 TI - What do consultation-liaison (C-L) psychiatry and psychosomatic medicine (PM) have in common? PMID- 12728521 TI - [Treatment of psychiatric patients with serious physical co-morbidity: administrative model in Tokyo]. AB - In Japan, treatment of physical co-morbidity among psychiatric patients has been difficult in many aspects. Conflicts frequently arise when referrals are made from mental hospitals to general hospitals for many reasons, such as prejudice, lack of psychiatrists, and lack of psychiatric beds in general hospitals. Tokyo Metropolitan District, with over 12 million people in population, has suffered this problem, consequently leading to an establishment of a government-involved referral system in 1981. The government mediates between mental hospitals and designated treatment centers so that smooth transfers are made. After two decades of operation, the number of transferred patients via the system is on the increase. While this system still has many problems to be solved, it may become a model administrative system for other governments to apply and provide hints for effective treatment of med/psych cases. PMID- 12728522 TI - Length of stay for medically ill patients with depression. AB - Depression in physically ill patients is often encountered in consultation liaison psychiatry. This investigation examined the relationship between length of stay (LOS) and depression comorbidity. Physically-ill patients with depression stayed significantly longer that those without. This study suggested that the early detection and antidepressant treatment is important for physically ill patients with depression that could lead to the shortening of LOS. PMID- 12728523 TI - A new role for C-L psychiatry: from ad-hoc services to integrated service delivery. PMID- 12728524 TI - [Clinical study of sarcomatoid renal cell carcinoma]. AB - Patients with sarcomatoid renal cell carcinoma are rare and have poor survival. We evaluated 14 patients who had renal cell carcinoma with a sarcomatoid component between 1982 and 2000. There were 9 men and 5 women with a median age of 59.5 years (range 32 to 77). Seven patients had a tumor on the right side and 7 on the left side. Thirteen patients had some symptoms and 11 had metastases at the initial visit. Most of them were stage T4 and high nuclear grade cancer and showed elevated acute phase reactants. There were 7 patients followed by interferon therapy, and the cause-specific 5-year survival rate was less than 10%. We confirmed that renal cell carcinoma with a sarcomatoid component often showed local invasion, distant metastasis and poor prognosis. PMID- 12728525 TI - [Acute pulmonary thromboembolism and deep-venous thrombosis at post-operative period in urologic surgery]. AB - Acute pulmonary thromboembolism (APTE) and deep venous thrombosis (DVT) are categorized into venous thromboembolism. APTE is one of the most critical post operative complications because of its high mortality. Meanwhile, DVT is responsible for more than 80-95% of APTE cases. APTE occurred in 6 patients and DVT in 3 patients in the post-operative period during the past eleven years at our hospital. The median age of these patients, 5 men and 4 women, was 62 years (45-77). In many cases, the course of APTE showed a rapid progression. Four of the six patients with APTE died, and two of these four patients died within a few hours following the onset. Once it occurs, it is often lethal. Therefore, the prevention of thromboembolism is most important. In this study, we examined these 6 APTE cases and 3 DVT cases and discussed low dose heparin and intermittent sequential pneumatic compression (ISPC), prophylaxis against APTE in the post operative period. We have been using ISPC for the high-risk group of thromboembolism in the post-operative period since 1999. In addition to using ISPC, we have been administering low dose heparin since November 2001. These 3 cases of DVT occurred regardless of adoption of ISPC. However, there were no cases of APTE after adoption of ISPC, and no cases of DVT or APTE after adoption of ISPC and administration of low dose heparin. PMID- 12728526 TI - [A case of psoas abscess caused by Candida albicans]. AB - We report a rare case of psoas abscess caused by Candida albicans. A 59-year-old man with diabetes mellitis presented with right flank mass. Computed tomography (CT) showed a psoas abscess. Needle aspiration of the right flank mass revealed pus which was proved to be C. albicans by culture. The literature is briefly reviewed. PMID- 12728527 TI - [A case of CA19-9-positive retroperitoneal mucinous adenocarcinoma]. AB - We report a case of retroperitoneal mucinous adenocarcinoma in a 44-year-old woman, who was admitted to our hospital complaining of low grade fever. Computed tomography and magnetic resonance imaging demonstrated a cystic tumor located at the lower pole of left kidney with irregular wall, which was enhanced by contrast medium. Open surgery was performed. Macroscopically, the tumor was clearly separated from the left kidney, and diagnosed as retroperitoneal in origin. Histopathological diagnosis was mucinous adenocarcinoma of retroperitoneum and immunohistochemical staining was positive for CA19-9, but not for CA125. The patient had no recurrence for 19 months post-operatively. PMID- 12728528 TI - [Retrovesical dermoid cyst in an adult male]. AB - A 46-year-old man complaining of transient dysuria. After digital examination and ultrasonography, computed tomography, magnetic resonance imaging and angiography, retrovesical tumor was suspected and tumor resection was performed. There was no adhesion to the adjacent organs, and histopathological diagnosis was a dermoid cyst. The patient is being followed up at our hospital without severe complications and recurrence. This is the third report of retrovesical dermoid cyst in a male in the world. PMID- 12728530 TI - [Primary malignant melanoma of the female urethra: a case report]. AB - A 72-year-old female with a complaint of hemorrhagic leucorrhea was referred to our department. She had a black tumor on her urethral meatus suspected as a malignant melanoma. Urethrectomy, vulvectomy, partial vaginectomy, bilateral inguinal lymphadenectomy, and cystostomy were performed. Pathological diagnosis was malignant melanoma of the urethra. Local recurrence and lymph node metastasis were found 9 months later, and she has been given DAV-feron combined therapy. PMID- 12728529 TI - [Retroperitoneal fibrosis: report of two cases]. AB - In case 1, a 24-year-old man was admitted with left lumbago. Abdominal computed tomographic (CT) scan showed the huge mass (8 x 10 x 13 cm in diameter) in the left retroperitoneal space, which lack enhancement and had a irregular margin. Percutaneous needle biopsy guided by ultrasonography was performed. In case 2, a 74-year-old woman was admitted with right lower abdominal pain. Laboratory data showed the renal failure. Imaging study revealed bilateral hydronephroses and bilateral masses at both common iliac vessels. A retroperitoneoscopic biopsy was performed. We diagnosed both cases as retroperitoneal fibrosis and administered prednisolone for 10 months. The masses decreased in size and the symptoms disappeared in both cases. PMID- 12728532 TI - [Three cases of congenital urethral stricture in childhood]. AB - We report three cases of congenital urethral stricture in boys. They were 8, 7 and 5 years old. They complained of enuresis both day and night. Voiding cystography revealed bulbar narrowing (Cobb's Collar) in all cases, and vesico ureteral reflux (VUR) occurred in two cases (three ureters). Endoscopically this lesion showed ring-form stenosis just distal to the urethral sphincter, and incised by using an infantile urethrotome. After the operation, VUR resolved in two ureters and improved in one. In all cases, daytime enuresis resolved dramatically, and night enuresis became controllable. Congenital urethral stenosis in boys is an important clinical entity in pediatric urology, and is not such a rare disease. PMID- 12728531 TI - [A case of fibroepithelial ureteral polyp in an adult patient]. AB - A 24-year-old man complaining of asymptomatic gross hematuria was referred to our hospital for treatment of bilateral hydronephroses (left > right) diagnosed at a previous clinic. Radiological examinations demonstrated a filling defect at the left uretero-pelvic junction. Cytological examination of the urine was negative. With the diagnosis of suspected benign ureteral polyp, the patient underwent endoscopic-assisted partial ureterectomy and pyeloplasty through a 5 cm flank incision. Histological examination revealed that the villous tumor in the ureter was a fibroepithelial polyp. PMID- 12728533 TI - Microscopic pulmonary tumor embolism secondary to adenocarcinoma of the prostate. AB - We report a case of pulmonary tumor embolism involving multiple emboli from an unusual site, an adenocarcinoma of the prostate. A 78-year-old Japanese man was diagnosed with stage IV (1997 version of the TNM classification) moderately differentiated adenocarcinoma of the prostate in December 1997. He underwent bilateral orchiectomy and hormonal therapy with flutamide was started. The patient suffered from relapse in April 1998, and estramustine phosphate was administered as treatment for hormone-refractory prostate cancer. He noticed a dry cough in May 1998, and on June 13, he developed acute progressive dyspnea and was admitted to our hospital. Radiological findings, blood gas analysis, and clinical symptoms suggested pulmonary thrombosis. Despite anticoagulation and oxygen therapy, he remained severely dyspnoeic. He died of respiratory failure 4 days after admission. Autopsy confirmed dissemination of poorly differentiated adenocarcinoma of the prostate to the majority of the pulmonary muscular arteries. PMID- 12728534 TI - [Bilateral partial nephrectomy in a case of bilateral synchronous renal cell carcinoma]. AB - We report a case of bilateral synchronous renal cell carcinoma in a 66-year-old man, who underwent bilateral partial nephrectomy. He visited our hospital, complaining of left flank pain. Drip infusion pyelography showed a left ureteral stone and left hydroureteronephrosis. Computerized tomography revealed bilateral renal tumors. These tumors were small (< 2 cm), so bilateral partial nephrectomy and left ureterolithotomy were performed. The pathological examination showed that all tumors were renal cell carcinoma. No recurrence has been seen nine months after the operation. This is the 12th case in the Japanese literature reported as bilateral partial nephrectomy for bilateral synchronous renal cell carcinoma. PMID- 12728535 TI - [Statistics on operations at the Department of Urology, Nippon Telegraph and Telephone West Osaka Hospital during a ten-year period from 1992 to 2001]. AB - A 10-year clinical statistic survey was made on the operations performed at the department of urology, NTT West Osaka Hospital between 1992 and 2001. The total number of operations was 2,540, comprising 1,899 males and 641 females, and a total of 1,559 ESWL was performed. The number of operations per year was chronologically constant. Major operations were transurethral resection of prostate (299 cases). The number of operations for malignant tumors, for examples radical nephrectomy and enucleation for renal cancer, nephroureterectomy for upper urinary tract cancer and transurethral resection of bladder tumor, has increased since 1997 year after year. The number of ESWL has decreased since 1994. Renal transplantation was introduced in 1995, and laparoscopic adrenalectomy also in 1997. PMID- 12728536 TI - [Different effects of exercise training in patients with myocardial infarction with or without diabetes mellitus]. AB - OBJECTIVES: This study investigated whether myocardial infarction patients with diabetes mellitus had lower heart rate reserve to exercise by measuring the increment in heart rate (HR) divided by the increment of norepinephrine (NE) concentration from rest to peak exercise (delta HR/log delta NE). The difference in exercise training effects was also investigated. METHODS: The 148 patients after myocardial infarction were divided into two groups, the DM group (n = 34) and the non-DM group (n = 114). Cardiopulmonary exercise testing was performed in each subject at 1 and 3 months after the onset. Blood samples were taken at rest and immediately after peak exercise, rest brain natriuretic peptide, rest and peak norepinephrine were analyzed. Exercise training was performed from 1 to 3 months after the onset. RESULTS: Resting heart rates were significantly higher in the DM group than in the non-DM group both at 1 and 3 months although peak heart rates were not significantly different. Peak oxygen uptake were lower in the DM group both at 1 and 3 months after onset of myocardial infarction compared to the non-DM group. End-tidal carbon dioxide pressure was lower and the rate of increase of minute ventilation to carbon dioxide output was higher in the DM group. Plasma brain natriuretic peptide was higher in the DM group. delta HR/log delta NE was 19.4 +/- 4.0 in the DM group and 22.2 +/- 5.6 in the non-DM group (p < 0.01), and increased in only the non-DM group. delta HR/log delta NE was more closely correlated with peak oxygen uptake in the DM group than in the non-DM group. CONCLUSIONS: Impaired response to exercise training may be caused, in part, by impaired heart rate reserve to exercise in patients with diabetes mellitus. PMID- 12728537 TI - Hemodynamics during chronic amiodarone administration in Japanese patients with idiopathic dilated cardiomyopathy and ventricular arrhythmia: a retrospective study. AB - OBJECTIVES: Nonischemic heart disease, especially idiopathic dilated cardiomyopathy, is relatively common among Japanese patients receiving amiodarone for concomitant ventricular arrhythmia, but the hemodynamic effects of amiodarone in these Japanese patients are unclear. The hemodynamic changes during chronic amiodarone administration were retrospectively studied in patients with idiopathic dilated cardiomyopathy and ventricular arrhythmia. METHODS: Fifty-two patients [42 males, 10 females, 53 +/- 2 years (mean age +/- SE)] with ventricular tachyarrhythmia and idiopathic dilated cardiomyopathy with left ventricular ejection fraction of 27 +/- 1% (mean +/- SE) were treated with 200 400 mg daily of oral amiodarone as the loading dose for the initial 14 days and 100-200 mg daily maintenance dose for a further 6 months. No patients were taking beta-blockers or positive inotropic drugs. Echocardiographic examination was performed before (baseline), at week 2 and at month 6 of amiodarone therapy. Twenty four-hour Holter monitoring during the same time period was also performed in 34 patients. Seventeen patients underwent right heart catheterization before and at week 2. RESULTS: Echocardiographic measurements showed no significant change in left ventricular end-diastolic dimension, although there was a slight increase in fractional shortening from 16 +/- 1% to 19 +/- 1% (p < 0.05) and 18 +/- 1% (mean +/- SE) (p < 0.01) at week 2 and month 6 of amiodarone therapy, respectively. Amiodarone markedly reduced the mean heart rate and the frequency of premature ventricular complexes on ambulatory monitoring. The cardiac index did not change and the pulmonary capillary wedge pressure tended to decrease slightly at week 2 in the 17 patients who underwent catheterization. CONCLUSIONS: This retrospective study showed no worsening of the hemodynamic state during chronic amiodarone administration in Japanese patients with idiopathic dilated cardiomyopathy and ventricular arrhythmia. PMID- 12728538 TI - [Myocardial systolic function of the left ventricle along the long axis in patients with essential hypertension: a study by pulsed tissue Doppler imaging]. AB - OBJECTIVES: Myocardial contractility of the left ventricle along the long axis in hypertensives is not well characterized. The systolic velocities of the left ventricular myocardium along the long axis were measured by pulsed tissue Doppler imaging in patients with mild to moderate essential hypertension. The relationships between the systolic velocity of left ventricular myocardium along the long axis and the blood pressure, and the left ventricular geometry were investigated. METHODS: The study included 60 untreated hypertensive patients (hypertension group) and 59 age-matched healthy subjects (control group). M-mode echocardiograms were recorded, and the relative wall thickness, left ventricular mass index and left ventricular end-systolic stress were calculated. The peak systolic velocities of the left ventricular posterior wall motion (Sw) were measured by pulsed tissue Doppler imaging. RESULTS: The Sw was significantly lower in the hypertension group than in the control group (8.3 +/- 1.9 vs 9.2 +/- 2.0 cm/sec, p < 0.05). The Sw was correlated inversely with systolic blood pressure (r = -0.31, p < 0.005), diastolic blood pressure (r = -0.25, p < 0.0001), interventricular septal thickness (r = -0.41, p < 0.0001), left ventricular posterior wall thickness (r = -0.39, p < 0.0001), relative wall thickness (r = -0.33, p < 0.001), and left ventricular mass index (r = -0.37, p < 0.001) in all subjects. CONCLUSIONS: The systolic velocity of the left ventricular myocardium along the long axis is decreased in patients with mild to moderate essential hypertension, and is negatively correlated with blood pressure and the severity of left ventricular concentric hypertrophy. PMID- 12728539 TI - Long-term cardiovascular effects of insulin sensitizer troglitazone on non diabetic individuals with insulin resistance: double blind, prospective randomized study. AB - OBJECTIVES: The study was undertaken to assess the long-term cardiovascular effects of troglitazone on non-diabetic individuals with insulin resistance. BACKGROUND: It has been suggested that treatment with troglitazone or similar insulin-sensitizing agents may prevent cardiovascular complications in non diabetic individuals with insulin resistance. However, the long-term cardiovascular effects of these agents on non-diabetic individuals remain to be defined. METHODS: A total of 137 African-American offspring of type 2 diabetic parents, with normal glucose tolerance and insulin resistance, were randomly divided to receive troglitazone 200 mg/day (n = 40), or placebo (n = 97) for 24 months. Brachial artery blood pressure (sphygmomanometry); aortic pulse wave velocity (carotid to femoral artery, Doppler), left ventricular diameters and mass (echocardiography); ascending and abdominal aortic distensibility (echocardiography, blood pressure); and metabolic and lipid profile were assessed at baseline, 12, and 24 months after randomization (delta 12, delta 24 respectively). RESULTS: The pulse wave velocity increased significantly in the troglitazone group compared to placebo group (p < 0.05). Changes from baseline in the troglitazone group were significant (delta 12 = 1.09 +/- 0.36 m/sec, delta 24 = 2.08 +/- 0.45 m/sec, ANOVA p < 0.001), while pulse wave velocity remained unchanged in the placebo group. This increase in pulse wave velocity is consistent with a decrease in the elastic properties of the aorta. CONCLUSIONS: Long-term administration of troglitazone to non-diabetic African-Americans with insulin resistance was associated with a decrease in the elastic properties of the aorta. Long-term therapy with troglitazone or similar agents for the prevention of cardiovascular complications in non-diabetic individuals with insulin resistance has to be critically evaluated. PMID- 12728540 TI - [Comparison of the efficacies of disopyramide, cibenzoline and aprindine for the termination of paroxysmal and persistent atrial fibrillation in elderly and non elderly patients]. AB - OBJECTIVES: The relationship between the efficacy of the anticholinergic action of disopyramide, cibenzoline and aprindine and age was examined in patients with paroxysmal and persistent atrial fibrillation. METHODS: This prospective, randomized study included 278 patients (200 men, 78 women, mean age 61 +/- 11 years) divided into two groups; the non-elderly group (age below 60 years) and the elderly group (age over 60 years). Successful termination was defined as conversion of sinus rhythm within 30 min of intravenous administration of 50 mg disopyramide (n = 91), 70 mg cibenzoline (n = 93) or 100 mg aprindine (n = 94) in this prospective and randomized study. RESULTS: No statistically significant difference was found in patient characteristics between the three agents. 1) The rate of conversion to sinus rhythm after disopyramide administration in the non elderly group(37.8%) was significantly higher than that in the elderly group (17.4%, p = 0.0361). 2) The rate of conversion to sinus rhythm after cibenzoline administration in the non-elderly group (62.2%) tended to be greater than that in the elderly group (43.8%, p = 0.0972). 3) The rate of conversion to sinus rhythm after aprindine administration in the non-elderly group (25.6%) was not significantly higher than that in the elderly group (18.2%, p = 0.4474). CONCLUSIONS: The anticholinergic action of antiarrhythmic agents has an effect on successful termination in non-elderly patients with paroxysmal and persistent atrial fibrillation. PMID- 12728541 TI - Diastolic musical murmur changing with cardiac cycle. Aortic valve prolapse due to myxomatous degeneration. PMID- 12728542 TI - Large hiatus hernia compressing the heart and impairing the respiratory function. PMID- 12728543 TI - [Epidemiological Studies on extremely low frequency electromagnetic fields and childhood leukemia, and preventive actions based on precautionary principles]. PMID- 12728544 TI - [Current status of nuclear neuroimaging]. PMID- 12728546 TI - Surface tension measurements in microgravity and their relevance to Marangoni convection. AB - The physical properties of liquids, such as surface tension, Marangoni convection and surface segregation are important in many technical fields and are of fundamental interest. However, inaccuracy of experiments has prevented us from fully understanding these phenomena. Recent advances in experimental techniques, such as the containerless technique and the utilization of microgravity, are described. PMID- 12728545 TI - Endophthalmitis with Klebsiella pneumoniae liver abscess. AB - Endogenous endophthalmitis is a rare, but devastating complication of septicemia. The prognosis of maintaining visual acuity in patients with septic endophthalmitis is poor in spite of an early diagnosis and the timely start of conventional therapeutic procedures because the intravitreous drug concentration remains low after the systemic administration of antibiotics due to the blood ocular barrier. We treated an elderly female patient with endogenous endophthalmitis complicated with disseminated intravascular coagulation associated with a Klebsiella pneumoniae liver abscess. Endophthalmitis developed rapidly and we thus had to perform an enucleation of both eyeballs even though we made an early diagnosis and performed liver abscess drainage as well as the prompt systemic and subconjunctival administration of antibiotics. Our experience in treating this case emphasizes the need to perform the timely intravitreous infusion of antibiotics with a support therapy consisting of the systemic and subconjunctival administration of antibiotics for endogenous endophthalmitis associated with a Klebsiella pneumoniae liver abscess. PMID- 12728547 TI - Alkyl monolayer passivated metal-semiconductor diodes: 2: Comparison with native silicon oxide. AB - To understand the electrical properties at passivated metal-semiconductor interfaces, two types of mercury-insulator-silicon (n-type) junctions, Hg?C10H21 Si and Hg?SiO2-Si, were fabricated and their current-voltage and capacitance voltage characteristics compared. Both of them exhibited near-ideal rectifying characteristics with an excellent saturation current at reverse bias, which is in contrast to the previously reported ohmic behavior of an unmodified mercury silicon junction. The experimental results also indicated that the n-decyl monolayer passivated junction possesses a higher effective barrier height, a lower ideality factor (that is, closer to unity), and better reproducibility than that of native silicon oxide. In addition, the dopant density and build-in potential, extracted from capacitance-voltage measurements of these passivated mercury-silicon junctions, revealed that alkyl monolayer derivatization does not alter the intrinsic properties of the silicon substrate. The calculated surface state density at the alkyl monolayer?silicon interface is lower than that of the silicon oxide?silicon interface. The present study increases the possibility of using advanced organic materials as ultrathin insulator layers for miniaturized, silicon-based microelectronic devices. PMID- 12728548 TI - High lithium ionic conductivity in the lithium halide hydrates Li3-n(OHn)Cl (0.83 < or = n < or = 2) and Li3-n(OHn)Br (1 < or = n < or = 2) at ambient temperatures. AB - Lithium ionic conductivity and phase transitions in a series of lithium halides hydrates and hydroxides with general formula Li3-n(OHn)X (0.83 < or = n < or = 2; X = Cl,Br) were studied using impedance measurements and 1H and 7Li NMR spectroscopy. All compounds studied in this work crystallize in the antiperovskite structure or are closely related to this structure type. With the exception of LiCl. H2O, all compounds with integer lithium content exhibit good lithium ionic conductivity in their high temperature cubic phases above T = 33 degrees C. Lithium doping of samples LiX.H2O and Li2(OH)X leads to a suppression of the phase transition into the noncubic phases and the good ionic conductivity is extended down to lower temperatures (T < 0 degree C). Thus, lithium doping of the lithium halide hydrates provides a promising tool for tailoring the ionic conductivity at ambient temperatures to its optimum value. PMID- 12728549 TI - OLED and PLED devices employing electrogenerated, intramolecular charge-transfer fluorescence. AB - The light generating mechanism of a series of light emitting diodes with electron donor-bridge-acceptor systems (D-b-A) as the emitting species was examined by constructing model diodes based on small organic molecules (OLEDs) as well as on molecularly doped electroactive (poly-N-vinylcarbazole, PVK) and insulating (polystyrene, PS) polymers (PLEDs). The direct electrogeneration of an intramolecular charge-transfer (CT) fluorescence of the donor-bridge-acceptor systems occurred readily in OLED devices with a D-b-A system as the emissive layer. In diodes with PS as the host matrix, hole-injection and electron injection occurred directly in the D-b-A molecules residing close to the anode and the cathode, respectively. In the PVK diodes, hole-injection occurred primarily into PVK and the positive charge carrier was subsequently trapped on the D-b-A molecule, whereas electron-injection at the cathode side occurred directly into the D-b-A molecules. Charge-hopping between neighboring molecules then occurred until a hole and electron resided on the same molecule, which is equivalent to the formation of the CT excited state, and which finally relaxed by intramolecular charge recombination under the emission of CT fluorescence. PMID- 12728550 TI - On the temperature--pressure free-energy landscape of proteins. AB - We studied the thermodynamic stability of a small monomeric protein, staphylococcal nuclease (Snase), as a function of both temperature and pressure, and expressed it as a 3D free-energy surface on the p,T-plane using a second order Taylor expansion of the Gibbs free-energy change delta G upon unfolding. We took advantage of a series of different techniques (small-angle X-ray scattering, Fourier-transform infrared spectroscopy, differential thermal analysis, pressure perturbation calorimetry and densitometry) in the evaluation of the conformation of the protein and in evaluating the changes in the thermodynamic parameters upon unfolding, such as the heat capacity, enthalpy, entropy, volume, isothermal compressibility and expansivity. The calculated results of the free-energy landscape of the protein are in good agreement with experimental data of the p,T stability diagram of the protein over a temperature range from 200 to 400 K and at pressures from ambient pressure to 4000 bar. The results demonstrate that combined temperature--pressure-dependent studies can help delineate the free energy landscape of proteins and hence help elucidate which features and thermodynamic parameters are essential in determining the stability of the native conformational state of proteins. The approach presented may also be used for studying other systems with so-called re-entrant or Tamman loop-shaped phase diagrams. PMID- 12728551 TI - An Ab initio study on the mechanism of the atmospheric reaction NH2 + O3-->H2NO + O2. AB - The atmospheric reaction NH2 + O3-->H2NO + O2 has been investigated theoretically by using MP2, QCISD, QCISD(T), CCSD(T), CASSCF, and CASPT2 methods with various basis sets. At the MP2 level of theory, the hypersurface of the potential energy (HPES) shows a two step reaction mechanism. Therefore, the mechanism proceeds along two transition states (TS1 and TS2), separated by an intermediate designated as Int. However, when the single-reference higher correlated QCISD and the multiconfigurational CASSCF methodologies have been employed, the minimum structure Int and TS2 are not found on the HPES, which thus confirms a direct reaction mechanism. Single-reference high correlated and multiconfigurational methods consistently predict the barrier height of the reaction to be within the range of 3.9 to 6.6 kcal mol-1, which is somewhat higher than the experimental value. The calculated reaction enthalpy is -67.7 kcal mol-1. PMID- 12728552 TI - Hydration and ion pairing in aqueous sodium oxalate solutions. AB - Dielectric spectra have been measured for aqueous sodium oxalate solutions up to the saturation concentration (0.04 < or = c[mmol L-1] < or = 0.25) at 25 degrees C over the approximate frequency range 0.2 < or = v [GHz] < or = 20. The spectra exhibit a process at about 1 GHz associated with the presence of ion pairs, in addition to the dominant solvent relaxation process at about 18 GHz. Detailed analysis of the solvent dispersion amplitude indicates that the oxalate ion is highly hydrated but that its solvation sheath is "fragile", decreasing quickly with increasing solute concentration. The NaOx(aq)- ion pair is shown to be of the double-solvent-separated (2SIP) type, with an infinite dilution association constant KA = 1.04 +/- 0.02. Analysis of the ion-pair relaxation time as a function of solute concentration gave rate constants for the formation (k12 = (7.3 +/- 0.4) 10(9) L mol-1 s-1) and dissociation (k21 = (6.7 +/- 0.5) 10(8) s-1) of the ion pair. These values are reasonably close to the diffusion-controlled values predicted by the Eigen theory, consistent with a 2SIP structure for the ion pair. PMID- 12728553 TI - Phase behavior and structural characterization of coexisting pure and mixed clathrate hydrates. PMID- 12728554 TI - Quasiharmonic vibrations of water, water dimer, and liquid water from principal component analysis of quantum or QM/MM trajectories. PMID- 12728555 TI - Magnetization transfer from laser-polarized xenon to protons with spin-diffusion quenching. PMID- 12728556 TI - Analysis of scanning tunneling spectroscopy experiments from first principles: the test case of C60 adsorbed on Au(111). PMID- 12728557 TI - Single-photon hot band absorption induced anti-stokes luminescence of rhodamine 101 in mesostructured thin films. PMID- 12728558 TI - Ab initio calculation of parity-violating chemical shifts in NMR spectra of chiral molecules. PMID- 12728560 TI - [Urologic complications in pelvic injuries]. AB - OBJECTIVE: Urological complications of pelvic fractures include in particular rupture of the urinary bladder (RUB), injury of the posterior urethra (IPU) and erectile dysfunction (ED). The authors present their own group of patients and in particular the diagnostic and therapeutic algorithm in IPU. MATERIAL AND METHODS: In the Plzen Faculty Hospital in 1/1998 to 8/2002 a total of 19 patients were treated with serious urological complications of pelvic fractures--9x RUB, 11x IPU (once with RUB). RUB was in one instance intraperitoneal, in the remainder extraperitoneal. RESULTS: IPU was without dislocation 6x, with dislocation 5x. Primary "realignment" of the urethra was made in 6 patients (in dislocations and in concurrent rupture of the bladder). In the remaining 5 an epicystostomy was established. In 4 after an interval of 3 months a posterior resection urethroplasty was made because of a distraction defect. One patient with a distraction defect was referred to the urological department of the catchment area and in another patient after-treatment is planned. Severe ED developed in 6 IPU of 10, always in dislocations of the urethra. In one patient we lack information on erections. CONCLUSION: When IPU is suspected (urethrorhagia), dislocation of the prostate on examination p.r.) ascendent urethrography and IVU are essential. Do not catheterize before completed examination. Then needle epicystostomy is performed, in major dislocations of the urinary bladder or in associated injuries primary "realignment" of the urethra open on a catheter or endoscopically. In distraction defects after 12 weeks a posterior resection plastic operation follows. PMID- 12728561 TI - [Effect of histologic classification on surgical treatment of peripheral nerve tumors]. AB - The influence of histological classification on the options of the peripheral nerve tumors surgery Peripheral nerve sheath tumors (PNST) constitute significant part of all tumors of peripheral nerves. In 1998-2000 years we have operated 7 such patients--3 females, 4 males in our department. The definite diagnosis was made by histology. Total removal was performed in all patients. The nerve continuity was preserved in 4 cases (3x neurofibroma, 1x schwannoma) and nerve had to be disconnected in 3 cases (1x neurobibroma, 1x plexiform neurofibroma as a part of von Recklinghausen disease (VRD), 1x malignant schwannoma). Good neurological outcome was in patients with preserved nerve continuity. Out of 3 patients with the need of nerve disconnection, graft and complete reinervation was performed in case of benign neurofibroma, but in the patient with von Recklinghausen disease and in the patient with malignant schwannoma the permanent leasion of nerve remained. We can conclude that histological classification directly predetermine the options of the peripheral nerve tumors surgery. PMID- 12728559 TI - [Cranial nerve vascular compression syndromes and neurogenic hypertension]. AB - The presented paper is a review of syndromes which till recently had an obscure cause and still have attributes such as "primary" or "essential". Their common denominator is vascular compression of the roots of the appropriate cephalic nerves. The syndromes include trigeminal neuralgia, glossopharyngeal neuralgia, hemifacial spasm, some forms of tinnitus and vertigo, exceptionally toricollis. Vascular compression of the ventrolateral oblongata on the left leads to neurogenic hypertension. The author discusses the state of knowledge on the aetiology of syndromes, the clinical and laboratory research of which contributed some findings on the physiology and pathophysiology of the mentioned conditions. At the same time the authors present their own experience with the therapeutic method of syndromes--microvascular decompression. PMID- 12728562 TI - [Topical fibrinolysis with streptokinase in coagulated hemothorax]. AB - Local fibrinolysis using streptokinase was used in two patients with an organizing extensive haemothorax. In both patients the coagula were dissolved and the pleural cavity was effectively evacuated by means of a drain without the necessity of an extensive surgical intervention. The authors did not record any side-reactions to the local administration of streptokinase. In the authors' opinion proper timing is important for achieving a satisfactory effect of local fibrinolysis. Fibrinolysis cannot be used in case of clinical signs of proceeding haemorrhage or closely after its termination. PMID- 12728564 TI - [Benign clear cell tumors of the lung--sugar tumors]. AB - The authors present the case reports of rare benign clear cells tumors of the lung (CCLT) and drawn attention, that examination from frozen sections can be misleading and definitive histological verification can be done on basis of immunohistochemical analysis and can be different from frozen sections examination. PMID- 12728563 TI - [Diagnosis of bone micrometastases in non-small cell lung carcinoma--initial results]. AB - The objective was to elaborate a method of immunohistochemical diagnosis of bone micrometastases and to apply this procedure in operated patients with the non small cell form of lung cancer. By means of a cocktail of cytokeratin mixtures and individual cytokeratins the immunohistochemical profile of the primary pulmonary focus is assessed. Bone micrometastases are evaluated from the bone marrow of a sampled costal segment. A total of 30 bone marrow samples were examined. For further evaluation 17 patients remained in the group. Isolated segments with a marked positivity of cytokeratins were observed in four instances. The yield of bone marrow from costal segment is very good. The initial results are encouraging for further research. PMID- 12728565 TI - [Importance of neoadjuvant chemotherapy in the treatment of advanced bronchogenic carcinoma]. AB - BACKGROUND: The basic task of induction (neo-adjuvant) therapy is elimination of occult micrometastatic dissemination found in some cases already in localized stages of non-small cell pulmonary cancer (stage I-IIIA NSCLC). An equally important effect is also cytoreduction in primary tumours which have before the local intervention an intact vascular supply. A difficult problem remains the correct selection of patients who from the long-term aspect may profit from such a procedure. MATERIAL AND METHODS: The authors evaluated perspectively aspects of oncological treatment and circumstances of surgical intervention after induction chemotherapy in 81 and 87 patients resp. in stage IIIA NSCLC evaluated before initiated neo-adjuvant chemotherapy. RESULTS: Complete remission was recorded in 4.9%, partial remission in 50.6%, stabilized disease in 23.5% and progression in 21% patients. Down-staging was recorded in 26%, 70.3% patients were indicated for surgery. In the group of 87 patients operated after induction therapy pneumonectomies predominated--41 (46%), only one operated patient died within 30 days after surgery (1.1%), complications were neither frequent nor serious. The median of survival after radical resection is 26 months. CONCLUSIONS: Neo adjuvant chemotherapy by modern cytostatics is usually well tolerated and creates satisfactory conditions for successful complete resection. The operation proper may be more difficult but need not be associated with serious complications. By this treatment it is probably possible to influence long-term results not only in stage IIIA but to reduce also the risk of a later more remote metastatic dissemination in some patients operated in lower stages of lung cancer. Our present aim is to test parameters which will be able to predict possible failure of induction therapy, and seek factors predicting risk behaviour of the tumour also in lower stages (stage I and II TNM classification). PMID- 12728566 TI - [Acute arterial occlusions of the upper extremities--clinical data]. AB - The authors describe 30 cases of acute arterial occlusions of upper extremity during the last 10 years (1992-2001) in a group of 27 patients. In 27 cases the occlusions were caused by embolism and in 3 cases by acute thrombosis. In the group were 12 men and 18 women. The embolus was the most frequently located in brachial artery (cubital)--17 times (63%). In one case we were forced to perform amputation of left upper extremity for gangrene. Authors accentuate that early and adequate treatment lead to salvage of extremity in 80-100%. PMID- 12728567 TI - [Surgical treatment of chronic venous insufficiency--current views on prevention, diagnosis and therapy]. AB - The authors describe their experiences with the therapy of CVI and inform about the present state of the surgical knowledge about this topic. The classification is based on the last recommendations from Roma (2000, modified CEAP). The surgical therapy of CVI they've divided into surgery of superficial venous system, surgical therapy of perforators and possibilities of surgical approach in the therapy of deep veins pathology (reflux and obstruction). They emphasize complex therapy in case of each patient and recommend the use of compressive and medical therapy as permanent and all life lasting method as the best strategy in this group of patients. PMID- 12728568 TI - [Should fiber gastroscopy be standard in preoperative examinations for cardiac surgery?]. AB - We introduced a gastrofibroscopy into a standard preoperative protocol for all patients indicated to the cardiac surgery. AIM OF THE STUDY: Validation of our protocol. MATERIAL AND METHODS: 957 consecutive patients operated on from May 1995 to January 2002 were divided to group A--gastrofibroscopy only for patients with positive history or clinical signs of peptic ulcer (151 pts.) and group B- patients with gastrofibroscopy as a standard procedure (806 pts). RESULTS: Significant difference in incidence of peptic ulcer complications between groups (A--5.9% versus B--1.1%, p < 0.05). Related mortality was 1.3% in-group A versus 0% in-group B. Positive history of peptic ulcer had only two patients (11% of 18) with postoperative complication. In-group B we found high incidence of peptic defects (B--35% versus A--9%, p < 0.01) requiring treatment by gastroenterologist. Majority (230 of 281-82%) of pts. with preoperative positive defect had negative history of a peptic ulcer. CONCLUSIONS: Current tendency to perform gastrofibroscopic examination only in patients with positive history of peptic ulcer is not able to detect "silent" lesions. Perioperative treatment with H2 blockers fails to prevent the manifestation of peptic ulcer complication. Gastrofibroscopy should be a routine part of the preoperative protocol in cardiac surgery. PMID- 12728569 TI - [Comments on the article "Comparison of the cost effectiveness of treatment of acute myocardial infarct with primary angioplasty and thrombolysis" by Golan et al]. PMID- 12728570 TI - [The search for the best reperfusion therapy in the treatment of acute myocardial infarct. An editorial on the article by Golan L. et al: "Comparison of cost effectiveness of treatment of acute myocardial infarct with primary angioplasty and thrombolysis"]. PMID- 12728571 TI - [Role of Chlamydia in atherosclerosis]. PMID- 12728572 TI - [Diagnostic markers of alcohol abuse]. PMID- 12728573 TI - [Comments on the article by Dastych M. et al: "Diagnostic value of carbohydrate deficient transferrin, gamma-glutamyltransferase and mean erythrocyte volume as laboratory markers of chronic alcohol abuse"]. PMID- 12728574 TI - [B-type natriuretic peptide in the differential diagnosis of dyspnea: a useful aid or meaningless ornament? (see the article by Stejskal D. et al: "Personal experience with determination of NT-proBNP in clinical practice" in this issue of Vnitrni Lekarstvi]. PMID- 12728575 TI - [Comparison of the cost effectiveness of treatment of acute myocardial infarct with primary angioplasty and thrombolysis ]. AB - INTRODUCTION: Primary angioplasty (PTCA) or intravenous thrombolysis are the recommended treatment of acute myocardial infarction. According to results of clinical investigations however primary PTCA provides a more favourable short term as well as long-term prognosis. As this method is much more expensive we were interested in its cost-effectiveness as compared with cheaper intravenous thrombolysis. METHODS: We constructed an decision analysis model (programme DATA 3.5, TreeAge Software) to compare the strategy of primary PTCA and intravenous thrombolysis in acute myocardial infarction. Probabilities of clinical outcomes were obtained from a long-term randomized clinical trial (Zijlstra et al. NEJM, 1999). The relative risk of death in PTCA was 0.54, rehospitalization 0.52 and reinfarction 0.27. The costs of PTCA (100,000,- crowns), of streptokinase thrombolysis (4000,- crowns) and hospitalization (2820,- crowns) were estimated from costs of the catheterization laboratory and information obtained from health insurance companies. We assumed that the subsequent costs of treatment and quality of life after the first infarction were the same in both strategies. In patients with reinfarction we anticipated a reduced quality of life (coefficient of life quality 0.9). The average effect of treatment and costs of both strategies were evaluated in the course of five years. As an acceptable cost effectiveness (ratio of difference in costs and effect) we considered costs up to 200,000,- crowns per one gained year of life. RESULTS: In the basic analysis we revealed that after 5 years the strategy of primary PTCA is more expensive (125,000,- crowns vs. 4500,- crowns) but has a greater effect, i.e. a longer life span (4.38 vs. 3.81) adjusted to quality of life). The cost-effectiveness (ratio of difference in costs and effect) expressing the costs of one gained year of life when using primary PTCA as compared with thrombolysis was despite the high cost of PTCA acceptable and amounted to 140,350,- crowns. Analysis of the sensitivity of the model confirmed the stability of favourable cost-effectiveness within a wide range of costs and therapeutic effect. CONCLUSION: Primary PTCA is in acute myocardial infarction a cost-effective strategy) providing effect for an acceptable cost) despite the markedly higher costs of the procedure. PMID- 12728576 TI - [Comparison of the occurrence of positive and negative vascular remodeling in patients with stable vs unstable angina pectoris]. AB - BACKGROUND: The vascular remodelling refers to the increase or decrease in EEM (external elastica membrane) area that occurs during development of atherosclerosis. The positive remodeling, which was thought only as a compensatory factor during atheroma development, was also found as a one of the main features of unstable plaque. The intravascular ultrasound is very good tool to measure different type of arterial remodelling. These findings correlate with histologic post-mortem specimen with excellent results. AIM OF THE STUDY: To correlate the remodelling index from patients suffered from stable angina pectoris (SAP) and from patients suffered from unstable angina (UA). METHOD: We performed IVUS in 51 patients. In the group of patients with unstable angina were included patients with worsening angina symptoms in last six weeks or with angina in rest. We used IVUS--endosonics In-Vision with 30 MHz probe Awanar with mechanical pull-back. We studied the occurrence of positive remodelling, negative remodeling (an index that describes remodeling is expressed as: lesion EEM CSA/reference EEM CSA. If the lesion EEM area is greater than the reference EEM area, positive remodelling has occurred, and the index will be > 1.0. If the lesion EEM area is smaller than the reference EEM area, negative remodelling has occurred, and the index will be < 1.0). RESULTS: We performed the intravascular ultrasound in 51 patients, 22 patients (43.1%) with unstable angina (UA) and in 29 patients (56.9%) stable angina (SAP). The positive remodelling was found in 14 patients (63.6%) in UA group vs. 5 patients (17.2%) in group with SAP, p < 0.05. The negative remodelling was found in 6 patients (27.3%) in UA group vs. 23 patients (79.3%) in SAP group, p < 0.05. CONCLUSION: Positive remodelling is more often found in patients with UA vs. in patients with SAP. Positive arterial remodelling is therefore one of the features of unstable atherosclerotic plaque. PMID- 12728577 TI - [Effect of chronic Chlamydia infection with non-specific inflammation on cardiovascular complications in acute myocardial infarct]. AB - It is known that local and systemic inflammatory processes play an important role in the genesis and development of atheroclerotic lesions and in the pathophysiology of acute coronary syndromes. This hypothesis is supported by findings of elevated parameters of the "inflammatory" reaction in the affected blood vessels but also in the blood of atherosclerotic patients. Known risk factors do not explain quite satisfactorily epidemiological cardiovascular phenomena and different manifestations of coronary heart disease. It is very probable that also Chlamydia pneumoniae is a risk factor. This assumption is based on evaluation of seroepidemiological data, examination of atherosclerotic plaques not only in humans but also in animal models with chlamydial infection. Based on retrospective and prospective evaluation of case-records the authors analyzed the incidence of cardiovascular complications in 83 patients with acute myocardial infarction (AIM), incl. 51 patients (31 men and 20 women, mean age 64.4 +/- 3.4 years who had a non-specific inflammation and chlamydial infection, and 32 patients (24 men and 8 women, mean age 64.7 +/- 3.6 years) who had chlamydial infections but no non-specific inflammation (in the blood). These patients were selected from all patients hospitalized during 1998-2001. When diagnosing acute myocardial infarction we applied WHO criteria, and the presence of at least two of three criteria was necessary: a history of prolonged (more than 20 min). stenocardia, electrocardiographic changes typical for ischaemia and/or necrosis and elevation of myocardial enzymes in serum, Non-specific inflammatory activity was present in patients (i.e. positive) if the following laboratory parameters were recorded: C-reactive protein > 5 mg/l assessed by the radial immunodiffusion method; fibrinogen > 4 mg/l assessed by the coagulation method according to Claus; leukocytes > 9.6 x 10(3)/microliter, leukocytes were counted automatically in a Coulter chamber; lymphocytes > 3.4 x 10(3)/microliter. Red cell sedimentation rate > 20 mm/hour. The activity was evaluated as positive when all parameters were elevated. The presence of chronic infection with Chlamydia pneumoniae was assessed qualitatively by antibody positivity (IgG) in serum using the microimmunoflurescent method (using a set from Labsystems Co.). The incidence of associated risk factors (obesity, smoking, diabetes, hyperlipidaemia and hypertension) is higher in the sub-group of patients with Chlamydia infections without inflammation, however, the difference is not statistically significant. The incidence of cardiovascular attacks was higher in the sub-group of patients with chlamydial infection and concurrent inflammation as compared with the sub-group of patients with chlamydial infection without inflammation. In case of re-infarction of the myocardium, a sudden cerebrovascular attack, death and arrhythmia the difference was statistically significant, while in case of cardiac failure and cardiogenic shock the difference was not significant. Patients with acute myocardial infarction with chlamydial infection and a concurrent non-specific inflammation had to be treated more often by combined (i.e. more intense) treatment, thrombolytic treatment, PTCA and surgery (bypass) of the coronary vessels as compared with patients with Chlamydia infections but without inflammation. The authors assume therefore that not only different risk factors but also the effect of non-specific inflammation and Chlamydia infection contribute towards the increased number of cardiovascular postinfarction complications. Therefore a therapeutic approach involving eradication of infection and suppression of the inflammatory reaction should be considered. PMID- 12728578 TI - [Diagnostic value of carbohydrate-deficient transferrin, gamma glutamyltransferase and mean erythrocyte volume as laboratory markers of chronic alcohol abuse]. AB - INTRODUCTION: Our research aimed at finding out values of carbohydrate-deficient transferin (CDT), gamma-glutamyltransferase (GGT) and mean corpuscular volume (MCV) for the purposes of future etiological diagnostics of alcohol neuropathy in thin fibres. METHODS: We examined the serum of 80 control subjects (50 women and 30 men), and the serum of 33 alcoholics (20 men and 13 women) with the daily consumption of more than 60 g alcohol in the course of the last four weeks. CDT was determined with the use of microcolumn separation after iron saturation followed by turbidimetric immunoassay (ChronoAlcoI. D., Sangui Biotech, Inc.) on Cobas-Mira analyser. CDT is expressed as a percentage of the total transferin. Senzitivity, specificity, positive likelihood ration (+LR), ROC and the area under the ROC curve were determined using statistical program MedCalc. RESULTS: The senzitivity, specificity and positive likelihood ratio (+LR) for CDT-%, respectively, were 82.6, 96.7 and 24.8 for men (cut off 2.2%), and 60.0, 88.0 and 5.0 for women (cut off 2.5%). The respective values for GMT were 95.7, 90.0 and 9.6 for men (cut off 0.64 mu kat/l), and 90.0, 80.0 and 4.5 for women (cut off 0.38 mu kat/l); for MCV 82.6, 96.7 and 24.8 for men (cut off 95.0 fL), and 80.0, 100.0 and 20.0 for women (cut off 97.2 fL). The area under the ROC curve for CDT %, GMT and MCV, respectively, were 0.940, 0.964 and 0.896 for men, and 0.829, 0.917 and 0.906 for women. CONCLUSION: In men, CDT-% and MCV showed the same values of the statistical parameters studied. GGT was more sensitive and less specific. In women, all the parameters studied presented a lesser diagnostic value, except for MCV with 100% specificity and +LR 20.0. PMID- 12728579 TI - [Personal experience with determination of NT-proBNP in clinical practice]. AB - INTRODUCTION AND OBJECTIVE: Recently in the literature information is found on estimation of natriuretic peptides in the differential diagnosis of dyspnoea. Because in the Czech Republic since the beginning of 2002 routine estimation of NT-proBNP is available (analyzer Elecsys 2010), the objective of our work was to find out whether it is possible to use in the everyday practice of a district hospital estimation of NT-proBNP to differentiate dyspnoea with affection of the heart muscle from other types of dyspnoea. METHOD: A group of 33 patients from the medical department of the Sternberk hospital was examined who attended on account of dyspnoea and lacked signs of acute coronary syndrome. All probands were diagnosed on the basis of defined criteria; according to the final diagnosis the patients were divided into three groups: group "LV" was formed by dyspnoic patients with organic affection of the left ventricle and signs of congestion in the lesser circulation, group "non-LV" was formed by patients where no organic affection of the left ventricle was found but other heart disease was present. Group "non-C" was formed by patients where a cardiac cause of dyspnoea was ruled out. In all patients on admission NT-proBNP was assessed. RESULTS: 33 probands were examined, 18 men and 15 women, mean age 74.5 years. 25 probands the dyspnoea was classified as dyspnoea with affection of one of the cardiac compartments [19 of them had signs of organic affection of the left ventricle (group "LV")]; in 6 probands no signs of organic left ventricular affection were found (group "Non LV"). The remaining 8 patients had no signs of any disease of the heart muscle, valves, septa, endocardium and pericardium (group "Non-C). The baseline values of NTpro-BNP were closely associated with the NYHA classification (grade II--median 55.3 pmol/l (469 ng/l, grade III--median 399.3 pmol/l (3384 ng/l), grade IV- median 724.7 pmol/l (6294 ng/l), the differences were statistically significant, p < 0.05). The dyspnoic probands with concurrent affection of some cardiac compartment (groups "LV" and "Non-LV") had a NT-pro BNP concentration significantly higher than probands without affection of the heart (group "Non-C") (median 589.5 pmol (4996 ng/l as compared with 62.9 pmol/l (533 ng/l, p < 0.01). In the group of probands with heart disease probands with affections of the left ventricle (group "LV") had significantly higher NT-proBNP values than subjects without affection of the LV and without any heart disease (groups "Non-LV" and "Non-C") (median 670.6 pmol/l (5683 ng/l) as compared with 187.5 pmol/l (1589 ng/l), p < 0.01). In hospitalized probands after treatment along with improved cardiopulmonary compensation also a significant drop of NT pro-BNP occurred (median 303 pmol/l (3967.7 ng/l to 211 pmol/l (2561 ng/l), p < 0.05). When looking for associations between anamnestic, laboratory and clinical data we found that the value of NT-proBNP is associated with dyspnoea with cardiac affection (groups "LV" + "Non-LV", correlation coefficient 0.48), with the left ventricular ejection fraction (correlation coefficient 0.52) and the baseline NYHA classification (correlation coefficient 0.36). In the examined group we did not find an association between NT-proBNP and age, sex, diabetes mellitus, hypertension, the presence of atrial arrhythmias, aortal stenosis, or the width of the left atrium. When using as cut-off for NT-proBNP 59 pmol/l (500 ng/l), the sensitivity of NT-proBNP for dyspnoea with affection of the cardiac compartments was 92% and the specificity 67%. CONCLUSION: Assessment of NT-proBNP is an important diagnostic acid in the differential diagnosis of dyspnoea. PMID- 12728580 TI - [Contrast nephropathy and its prevention]. AB - Contrast-induced nephropathy is one of the adverse events of diagnostic and therapeutic intravascular application of contrast agent. In general, the condition was defined as an increase in the serum creatinine concentration of more than 44 mmol/l or of more than 25% within 48 hours after the contrast agent administration. Other cause of creatinine increase should be excluded. Contrast induced nephropathy has been reported to be the third leading cause of acute nephropathy in hospitalized patients, occurring at a rate of 1-6% in unselected population and of 30-50% in high-risk patients. One year mortality can be as high as 45% in high-risk patient population. The most important risk factors are chronic renal insufficiency, diabetes mellitus and high volume of contrast agent. Clinical presentation is mostly asymptomatic, but in some patients acute renal failure with necessity of hemodialysis can occur. Prevention is underlying tool in reducing of contrast-induced nephropathy incidence. It is based on the identification of risk patients, stop of medication which can increase risk of contrast-induced nephropathy and proper hydratation of patients before, during and after the contrast agent administration. In high-risk patients, non-ionic and low-osmolarity contrast agent should be used. Several clinical studies testing different drugs to prevent contrast-induced nephropathy were performed, but no convincing result has been found. Promising substancies are N-acetylcysteine and fenoldopam. PMID- 12728581 TI - [Long-term effect of catheter ablation of ventricular tachycardia in adult patients with previous surgery of congenital heart defects. 2 case reports]. AB - Two case reports of catheter ablation of sustained monomorphic ventricular tachycardia in two adult patients after previous surgical correction of congenital heart disease are reported. In both patients catheter ablation led to long-term elimination of ventricular tachycardia during follow-up period of 38 and 20 months. PMID- 12728582 TI - [Teaching of pathologic physiology in the spirit of Dr. Frantisek Pora]. PMID- 12728583 TI - [The hemophagocytic syndrome]. PMID- 12728584 TI - [Editorial on the article by O. Valocik et al: "Effect of ACE gene polymorphism on left ventricular function in type 2 diabetes"]. PMID- 12728585 TI - [The stomach and bone metabolism]. PMID- 12728586 TI - [Right-sided infectious endocarditis--an uncommon but increasing problem]. PMID- 12728587 TI - [Dementia in the Czech Republic--views of an internist and geriatrician]. PMID- 12728588 TI - [Comment on the article by S. Krajcik: What are the possibilities of preventing dementia?]. PMID- 12728589 TI - [Editorial on the article by H. Svacinova et al: "Effect of walking exercise on aerobic capacity and blood lipids in type 2 diabetes]. PMID- 12728590 TI - [Long-term results of the UHKT-911 study of adult patients under 65 years of age with de novo acute myeloid leukemias without favorable karyotypes]. AB - Between February 1991 and April 1994 induction chemotherapy of 32 adult consecutive patients under 65 years with de novo acute myeloid leukemias (AML) was started in the study UHKT-911. They were 19 women and 13 men, aged 18-63 (median 44) years. Their AML were classified according to the FAB classification: 3 M0, 3 M1, 9 M2, 14 M4, 3 M5. Induction chemotherapy consisted of 1-2 cycles with 3-4 doses of daunorubicin (DNR) 45 mg/m2/d i.v. and 14 doses of cytosine arabinoside (Ara-C) 200 mg/m2 per 3-h infusion every 12 hours. After the treatment patients, not being in complete remission, got the HD cycle with 10 high-doses of Ara-C 2000 mg/m2 per 3-h infusion every 12 hours i.v. and DNR 45 mg/m2/d i.v. on days 4 and 5, then the EMi cycle composed of etoposide 100 mg/m2/d i.v. for 5 days and mitozantrone 10-12 mg/m2/d i.v. on days 1, 3 and 5. Complete remission (CR) was achieved in 25 of 32 (78%) patients after 1-3 cycles. Five patients died between days 5 and 24 of treatment of infections, two patients were resistant to 4 cycles of induction therapy and survived 8.4 and 13.5 months. Three patients chose allogeneic bone marrow transplantation in their 1st CR from their relatives. Two of them have been living in CR for 115 a 110 months since diagnosis, the third died of sepsis on the day 52 after transplantation. Two patients in CR died of infections after their 2nd. consolidation cycle. Twenty patients in CR completed 2-4 consolidation cycles (1-3 HD, 1 EMi). Median of their CR duration was 17.8 (2-117) months. Relapse appeared in 12 cases after 4.4 34.8 (median 12.5) months, 8 patients (6 women and 2 men, aged 29-63 years) have remained longer than 5 years in their 1st. CR. Cytogenetic examination of their bone marrow showed a normal karyotype in 4 cases, 1x 46,XX,del(1)(p32p34), 1x 46,XX,16p+, 1x 47,XX,+mar, 1x 46,XX,del(5)(q22q33). After 62 months in CR a pancytopenia with dysplastic bone marrow changes developed in one of them, probably a secondary myelodysplastic syndrome, lasting for further 33 months. Event-free survival at 5 years was 27.5% (8/29 patients), significantly better (p = 0.046) against 7.5% (3/40) patients treated without HD cycles in the years 1982 1987. The same difference was observed in 7.5-year overall survival (p = 0.036) between the two studies, when 3 of 6 patients 60-64 years old remain in their 1. CR. PMID- 12728592 TI - [Importance of measurement of carotid artery distensibility]. AB - Affections of the structure of the major blood vessels can be assessed non invasively by ultrasound. The authors assessed the thickness of the intima-media (IMT) and distensibility (D) of the carotid arteries in patients with ischaemic heart disease (IHD) and a cerebrovascular attack (CVA). Duplex sonography was used in 234 patients (145 patients with IHD and 89 patients with CVA) incl. assessment of IMT and D. 59 patients had type 2 diabetes and 64 had atrial fibrillations. D was evaluated according to Raneman's formula (mm/100 mm Hg). Diabetic subjects had a poorer D in the group with IHD (0.16 vs. 0.20, p < 0.05) as well as in the CVA group (0.14 vs. 0.17, p < 0.05) and the finding correlated with IMT. Insulin treatment as compared with PAD treatment did not influence D in a significant way (0.15 vs. 0.16, NS). Th presence of atrial fibrillation in patients with IHD did not affect the investigated parameters (IMT 0.72 vs. 0.74 mm, NS, D 0.19 vs. 0.18, NS). In patients with CVA, as compared with patients with a sinus rhythm, better IMT findings were recorded (0.67 mm vs. 0.79 mm, p < 0.05) and D (0.19 vs. 0.10, p < 0.05). The results indicate that IMT an D correlate indirectly and this is made more apparent by the presence of diabetes. In patients with CVA the different findings in relation to the presence of atrial fibrillation indicate the possibility to use these parameters to differentiate an embolic and thrombotic etiology of the attack. The results assembled by the authors do not support a better informative value of D as compared with IMT. PMID- 12728591 TI - [Effect of ACE gene polymorphism on left ventricular function in patients with type 2 diabetes]. AB - The authors evaluated the influence of the ACE gene polymorphism on left ventricular function in patients with diabetes type 2. In a group of 23 patients left ventricular mass, global and regional systolic function of the left ventricle as well as diastolic function was assessed from transmitral flow using two-dimensional echocardiography. In a subgroup of patients with DD (n = 7) and ID genotype (n = 16), no significant differences were found in age (p = 0.19), duration of diabetes (p = 0.46), level of HbA1c (p = 0.10), cholesterol level (p = 0.18), quantitative proteinuria (p = 0.39), systolic and diastolic blood pressure (p = 0.25, p = 0.40). No association was found between insertion deletion polymorphism of the ACE gene and observed echocardiographic parameters [left ventricular mass index (p = 0.43), EF (p = 0.46), wall motion index (p = 0.25), E wave (p = 0.14), A wave (p = 0.07), deceleration time of the E wave (p = 0.06), E/A (p = 0.07), flow propagation velocity (Vp) (p = 0.14) and E/Vp (p = 0.38)]. The presence of myocardial infarction, ischemic heart disease and hypertension had no association with ACE gene polymorphism (p = 0.53, p = 0.61 and p = 0.64). In conclusion, there is no association between ACE gene polymorphism and systolic and diastolic function of the left ventricle in this group of patients with diabetes type 2. PMID- 12728593 TI - [Myelotoxicity after systemic radionuclide therapy of painful bone metastases with 153Samarium-EDTMP ]. AB - AIM: The breast cancer and the prostate cancer are considered as the most frequently occurred metastatic tumors in bones. The bone pain appears in majority of patients with metastases. One possibility how to reduce or to remove this pain is the administration of the radiotherapy to the place of metastases. The present medicine disposes of two ways how to administer radiotherapy in the place of metastases: externally irradiation (loco-regional or half-body) or intravenous administration of bone-seeking therapeutic radiopharmaceuticals. MATERIAL AND METHOD: We introduced intravenous administration of 153Samarium-EDTMP in 57 patients (30 prostate cancer, 23 breast cancer, 4 renal cell carcinoma). Mean applied activity was 40 MBq per kg of patient's body weight. RESULTS: After 153Sm EDTMP administration we observed pain relief of various degree in 75% of patients for three months. 1 and 3 months after 153Samarium-EDTMP administration the following haematologic parameters changes were seen: a) 7% reduction in red cell count in one month after administration, 15% reduction after three months. b) 43% reduction in leukocyte count in one month after administration, 24% reduction after three months. c) 50% reduction in platelet count in one month after administration, 23% reduction after three months. In all three haematological profiles (red cells, leukocytes, platelets) we detected reduction of values after treatment with 153Samarium-EDTMP. This reduction culminates one month after administration and in three months leukocytes and platelets count was getting better. CONCLUSION: Myelosuppression was mild and temporary. No patients had grade 4 hematological toxicity and only three patients (5%) had grade 3 hematological toxicity (National Cancer Institute Common Toxicity Criteria). PMID- 12728594 TI - [The effect of walking exercise on aerobic capacity and serum lipids in type 2 diabetics]. AB - Regular physical activity can have a favourable impact on other risk factors of ischaemic heart disease (IHD) and associated diabetes (DM), such as obesity, hypertension, dyslipidaemia, insulin resistance and others. This important part of treatment of diabetes is frequently difficult to implement because of the lack of willingness ("adherence") of type 2 diabetics to practice regular exercise, and unequivocal data are lacking on the intensity of exercise which will influence effectively these risk factors and be at the same time safe, readily available and psychologically acceptable. The objective of the work was to find out whether walking, i.e. locomotor activity with a low to medium intensity can effectively influence parameters of aerobic capacity and blood lipids. The authors submit the results of two groups of type 2 diabetics. The experimental group B (n = 10, age 57 +/- 7 years, BMI 31 +/- 3, duration of DM 8 +/- 5 years) participated in a 12-week training programme of walking; at the beginning and at the end of this period indicators of aerobic capacity at the level of the anaerobic threshold (VO2ANP) were evaluated as well as at the level of the symptom limited maximum (VO2SL, TepO2SL), and the blood lipid levels. In the control group A (n = 6, age 58 +/- 7 years, BMI 32 +/- 4) indicators of aerobic capacity and blood lipids were assessed after a 12-week period of the usual habitual physical activity. In group B the 12-week walking training led to significant improvement of parameters of aerobic capacity at the level of the anaerobic threshold (ANP), oxygen pulse at the level of the symptom limits maximum (SL) and a significant reduction of total and LDL cholesterol. In the control group no significant changes occurred in aerobic capacity nor blood lipid values. The training programme where walking was selected as physical activity with a low to medium intensity can be considered suitable for everyday life of motivated patients with type 2 diabetics, preferably in the form of a domestic training programme. The prerequisite of success is its regular and frequent evaluation by health professionals. PMID- 12728595 TI - [Hemophagocytic syndrome (case report and literature review)]. AB - The haemophagocytic syndrome (HPS) is clinically characterized by fever, pancytopenia and hepatosplenomegaly. Usually it takes an acute course with a high mortality. The pathogenetic basis is inadequate activation of the immune system- in particular Th1-lymphocytes with subsequent overproduction of cytokines and extreme activation of macrophages with haemophagocytosis. The activated cells infiltrate organs, cause tissue damage and clinical manifestations of the syndrome. From the etiological aspect two forms exist: primary (familial) with autosomal recessive inheritance and the secondary form which forms a heterogeneous sub-group, caused as a rule by infection and/or a tumour. The prognosis seems somewhat more favourable in secondary forms. In treatment which is essentially the same for both forms, chemotherapy combined with immunosuppression proves useful, in more aggressive forms chemotherapy as used in the treatment of non-Hodgkin lymphomas. The only curative method is transplantation of haematopoietic stem cells which is also the treatment of first choice in the familial form of haemophagocytosis. In the submitted paper the authors present a review of contemporary knowledge on this treacherous and relatively rare entity. The haemopgagocytic syndrome should be always taken into account in the differential diagnosis of fever with an obscure etiology. The authors assume that the haemophagocytic syndrome is rarely considered in practice and therefore is usually inadequately diagnosed and thus not treated in time. In the conclusion the authors describe the case-records of a 26-year-old female patient with haemophagocytic syndrome which developed during pregnancy. PMID- 12728596 TI - [Atherosclerotic nephropathy in renal artery stenosis--from randomized studies to individualized therapy]. AB - Randomized trials in hypertensive patients with atherosclerotic renal artery stenosis (ARAS) mostly did not reveal any significant difference between antihypertensive treatment and revascularization (by angioplasty or bypass surgery) in their effects on blood pressure or glomerular filtration rate. This unexpected conclusion reflects a fact that in addition to potentially reversible ischemia, some other factors which are not eliminated by technically successful revascularization take part in the decrease of renal function in ARAS, including cholesterol microemboli from atherosclerotic plaques, secondary focal segmental glomerulosclerosis and hypertensive nephroangiosclerosis. Moreover, these changes have been also found in the contralateral kidney without any stenosis. Scintigraphic studies confirmed that the individual kidney function was not related to the presence of ARAS, i.e., the glomerular filtration rate in the stenotic kidney was often equal to, or paradoxically even better than that in the kidney with normal renal artery. This has obviously important consequences for the indication of revascularization which should be based on measurement of the individual kidney function rather than overall renal function. A conservative treatment of ARAS should comprise ACE inhibitors or angiotensin II receptor antagonists, statins and acetylsalicylic acid. The long-term effect of such treatment on the progression of atherosclerotic nephropathy is now being evaluated in randomized trials. PMID- 12728597 TI - [Plasminogen activator inhibitor-1 (PAI-1), ischemic heart disease and diabetes mellitus]. AB - The classic risk factors can only in part explain the increased incidence and extent of coronary artery disease. Plasminogen activator inhibitor type-1 (PAI 1), the main human regulator of endogenous fibrinolysis, is considered to be an important cardiovascular risk factor. The article discuss the current knowledge of the PAI-1 importance and role in the in the pathogenesis of atherosclerosis and mechanisms of its synthesis predominantly in patients with coronary artery disease associated with diabetes mellitus type 2. It explains contributions of PAI-1 to the pathophysiologic links between diabetes mellitus and cardiovascular diseases and shows current therapeutic interventions normalising PAI-1 activity, their beneficial effect on prognosis and course of coronary artery disease. PMID- 12728599 TI - [What are the possibilities of preventing dementia?]. AB - Population ageing makes issue of dementia prevention very important. Measures used in prevention of other diseases are also effective in prevention of dementia. Incidence of vascular dementia is decreased by effective treatment of hypertension, diabetes and anticoagulatory treatment in patients with atrial fibrillation. Nonsteroidal antiinflammatory drugs and statins reduce incidence of Alzheimer's disease. PMID- 12728598 TI - [Specific features of hematopoiesis and hemostasis in advanced age]. AB - The ageing process affects also haematopoiesis and haemostasis and these changes can have a far reaching effect on other organ systems and their function. The authors decided therefore to present a review on involutional changes on the formation and function of blood elements and the system of blood clotting. The review is divided into description of changes in the bone marrow and explanation of hitherto known facts on the pathophysiological background of these changes. Furthermore the authors deal with changes in the peripheral blood stream and their mutual relations with administered drugs, diseases under way, reactions to external stimuli such as immunization etc. In the second part of the review the authors describe changes of haemocoagulation, emphasize the tendency of hypercoagulation and the mutual relationship of the development of atherosclerotic changes and hypercoagulation trends and their pathophysiological background. Attention is paid to thromboembolic disease and its increasing risk in elderly patients incl. the possible late manifestation of congenital defects of coagulation inhibitors. The authors mention also the influence of lifestyle and dietary factors on the coagulation activity in old age. They analyze in detail possibilities and pitfalls of prophylaxis of thromboembolic disease in elderly patients and the risks of haemorrhagic complications of different operations. PMID- 12728600 TI - Protracted infection of pacemaker leads with development of large vegetations: three years follow up. Case report. AB - A case history of a patient with permanent pacemaker reports inaccurate diagnosis of inflammation at the site of operation wound and a protracted infection of pacemaker leads. Inadequate therapy led to protracted inflammation with the leads fixed in the superior vena cava and the development of large vegetations in the right heart. Finally, the condition was resolved by surgical explantation and long-term antibiotic therapy. This case history describes infectious complications related to the implantation of a permanent pacemaker and the role of echocardiography in diagnosis of vegetations and points out the surgical treatment as a method of choice. PMID- 12728601 TI - [Idiopathic aneurysm of the pulmonary artery]. AB - Aneurysm of pulmonary artery (PAA) is relatively rare clinical finding and appears mostly in association with significant cardiovascular or pulmonary abnormalities and is frequently caused by pulmonary hypertension. Its diagnosis is not difficult. However, guidelines for the treatment were not yet established. Patients with PAA are at risk for sudden death by a rupture of aneurysm. Therefore, surgical correction (graft replacement) is the method of choice. Nevertheless, it seems that certain group of patients with asymptomatic idiopathic aneurysm of pulmonary artery without presence of any serious cardiovascular or pulmonary abnormality can profit from conservative approach. We refer a 62-years old man with idiopathic PAA, in whom we did not indicate surgical correction. However, the long-term follow-up is necessary. PMID- 12728602 TI - Advocacy: your investment in medicine's future. PMID- 12728603 TI - HIPAA business associates--be aware of contracting responsibilities and wary of risks. PMID- 12728604 TI - IMS continues to advocate for change. PMID- 12728605 TI - "Jousting" revisited: don't fall into this liability trap. PMID- 12728606 TI - Competition still illusive in Iowa's concentrated health insurance market. PMID- 12728607 TI - Help is on the way! PMID- 12728608 TI - Tort reform--what about caps? PMID- 12728609 TI - Students with chronic illnesses: guidance for families, schools, and students. AB - Developed by a work group representing several nonprofit and federal health and education organizations, Students with Chronic Illnesses: Guidance for Families, Schools, and Students provides cross-cutting action steps to facilitate full participation in learning and other school activities by students with chronic diseases. Between 10% and 15% of children in the United States are affected by at least one chronic disease. This guide offers practical tips to help ensure that schools are responsive to the health needs of children with chronic illnesses. Children with chronic health conditions are more likely to miss days from school, need specialized health care, and require special education services or homebound teaching. School health advocates agree that meeting the needs of these children requires collaboration among the students, their families, school personnel, and community health care providers. Such efforts can help students establish better attendance, improve their alertness and physical stamina, and face fewer restrictions on physical activity at school and fewer medical emergencies. The guide addresses issues such as routine and urgent care needs of students, medication access and administration, and school personnel training to ensure compliance with applicable local, state, and federal regulations. While emphasizing the value of adopting general policies for supporting students with chronic conditions, the guide also encourages schools to develop and implement condition-specific and individualized protocols. The responsibilities of family members, school district officials, teachers and school administrators, and students are outlined to help each constituent fulfill these objectives. PMID- 12728610 TI - A survey of health promotion at international schools. AB - This study investigated health promotion efforts at international schools serving the education needs of expatriate communities abroad. Factors supporting the implementation of whole-school approaches to health promotion also were examined. Self-completed questionnaires were distributed by a combination of electronic and conventional mail. International school staff in 93 countries (n = 205) completed an adapted version of an instrument used for evaluating the Western Australian School Health Project (WASHP). This survey demonstrated usefulness of the WASHP instrument cross-culturally in a variety of school settings. The level of whole school approaches to health promotion in the participating international schools varied but tended to be low. Demographic characteristics of schools were not associated with differences in the level of health promotion, with the exception school size. School organizational factors support implementation of health promotion programs. PMID- 12728611 TI - A role for Community HealthCorps members in youth HIV/AIDS prevention education. AB - One-half of all new infections with the human immunodeficiency virus (HIV) and cases of acquired immunodeficiency syndrome (AIDS) occur in young people under age 25. While many markers for the spread of HIV/AIDS have been declining, the Centers for Disease Control and Prevention (CDC) reports no similar reduction in newly diagnosed HIV cases among youth. These findings point to the need for greater prevention and education efforts to help curb the spread of HIV/AIDS among the nation's youth. Community HealthCorps members working in community health centers provided HIV/AIDS prevention education to youth as part of their community service work. After HealthCorps members received training in the HIV/AIDS prevention curriculum, Be Proud! Be Responsible!, they recruited young people from their communities to participate in the program. HealthCorps members provided prevention education to students to ensure they acquired basic knowledge about HIV/AIDS on which to base personal decisions and actions. Staff of the Adolescent School Health Initiative of the National Association of Community Health Centers assessed the impact of the education on youth knowledge of HIV/AIDS, understanding of personal risk, their ability to refuse or negotiate safer sex, and proper use of condoms. This article highlights findings and summarizes lessons learned from the prevention education initiative. PMID- 12728612 TI - Variation in school health policies and programs by demographic characteristics of US schools. AB - To understand the relationship between demographic characteristics of schools and school health policies and programs, this study analyzed data from the School Health Policies and Programs Study (SHPPS) 2000. SHPPS 2000 provides nationally representative data on eight components of school health. Data were collected from school faculty and staff using onsite, computer-assisted personal interviews, then linked with extant data on school characteristics. Results from a series of regression analyses indicated that the presence of most policies and programs examined differed according to school type (public, private, or Catholic), urbanicity, school enrollment size, per-pupil expenditure, percentage of White students and, among high schools, percentage of college-bound students. No one type of school, however, was more likely than another type to have all key aspects of a school health program in place. Regardless of school characteristics, all schools are capable of implementing quality school health programs. PMID- 12728613 TI - Feasibility of school-based spirometry screening for asthma. AB - To determine the feasibility and value of spirometry in school-based asthma screening, spirometry testing was coupled with parent questionnaires in a school based asthma screening project. Children in grades five to eight of the Catholic school system in Rochester, Minn., performed spirometry with coaching and data acquisition by nurses trained for this activity. Most students completed three tests. For each student, the best test was selected for interpretation. Tests were considered technically unacceptable for screening purposes if the FEV1 was less than 85% and the curve showed evidence of cough, delayed start, poor initial effort, incomplete effort, or non-reproducibility. Students with acceptable tests and FEV1 < 85% as predicted for age, race, and BMI were classified as appropriate for referral for further evaluation of potential asthma. A sensitivity analysis was conducted using different FEV1 thresholds for referral. Children (119, 17.6% of all) with known asthma based on parent-completed questionnaire were not considered for referral. Of the remaining 557 students screened, 535 had technically acceptable tests, and 498 had normal spirometry performance. Using a threshold for referral of FEV1 < 85%, 37 children were candidates for referral for further evaluation of potential asthma. Only four (11%) of these also had questionnaire responses that made them candidates for referral. School-based spirometry screening for asthma is technically feasible but there is little overlap between those who are referral candidates based on spirometry data and those who are referral candidates based on parent-reported symptoms on screening questionnaires. Without further study, spirometry cannot be recommended for school-based asthma screening. PMID- 12728614 TI - Symptoms of nicotine dependence and other predictors of student smoking at school: implications for school smoking policy. AB - Students who violate school smoking policies present a difficult health and disciplinary issue for school officials. Professionals know little about the characteristics of students who smoke at school. In a prospective study of 679 students in two cities in central Massachusetts, researchers examined how nicotine dependence contributes to the problem of smoking at school. After three years of follow up, smoking at school was reported by 10.3% of students. Among subjects who admitted to smoking at school, 63% reported that symptoms of nicotine dependence preceded their smoking at school. After adjusting for other variables, student smokers with symptoms of nicotine dependence were nine times more likely to report smoking in school (OR 9.1, 95% CI 2.9-28.5) than were student smokers without symptoms. Smoking at school was more common among daily smokers and those who paid for their own cigarettes. Age, gender, race, and parental smoking status were not significantly associated with students' reports of smoking at school. These data suggest nicotine dependence as an important contributor to the problem of smoking at school, but not the only reason why students violate school smoking policies. Disciplinary action against students caught violating school smoking policies should be supplemented with an offer of treatment for nicotine dependence. PMID- 12728615 TI - Relationship between student illicit drug use and school drug-testing policies. AB - This report provides information about drug testing by American secondary schools, based on results from national surveys. The study provides descriptive information on drug-testing practices by schools from 1998 to 2001, and examines the association between drug testing by schools and reported drug use by students. School-level data on drug testing were obtained through the Youth, Education, and Society study, and student-level survey data were obtained from the same schools participating in the Monitoring the Future study. A relatively small percentage of schools (about 18%) reported testing students for drug use, with more high schools than middle schools reporting drug testing. Drug testing was not associated with students' reported illicit drug use, or with rate of use among experienced marijuana users. Drug testing of athletes was not associated with illicit drug use among male high school athletes. Policy implications are discussed. PMID- 12728616 TI - Surveyor group trained for 2003 pilot surveys on shared visions-new pathways. PMID- 12728619 TI - Infection control expert panel to review IC standards, survey process. PMID- 12728618 TI - JCAHO staff to perform unannounced surveyor mentoring visits during surveys. PMID- 12728620 TI - 2002 winners of the Ernest A. Codman individual award. PMID- 12728621 TI - Greater New York Hospital Association offers more insight on emergency management. PMID- 12728622 TI - Oxygen aids healing process. PMID- 12728623 TI - Great heights achieved for the Siobhan Rankin fund. AB - It is one year since the unexpected and tragic death of Siobhan Rankin, Chairman of NATN. It is timely then to publish this account of Marion Taylor and Lynn Graff's successful climb of Africa's highest mountain, Mount Kilimanjaro, that took five-and-a-half days and one-and-a-half days to come down. This article gives some insight into their experiences and challenges. PMID- 12728624 TI - Recovery nursing services. An evolution. AB - This article examines and charts the development of recovery services in the context of the social and scientific matrix of historical development. Such a reflection is important to gain an understanding of our future challenges and the direction that the perianaesthesia profession may take. PMID- 12728625 TI - Reading and critiquing research. PMID- 12728626 TI - Being bullied what an insight. AB - Bullying and harassment have occupied a prominent position in NATN workshops and publications recently. Challenging Behaviours in the Perioperative Environment identifies practices that fall within the definition of these socially and legally unacceptable activities, and provides advice and guidance to victims and those who use bullying or violent behaviours. In this article, the author draws on her own experience of being a victim and provides advice on how to identify and deal with bullying. PMID- 12728627 TI - [Mycelial wastes of penicillin fermentation as glue components]. AB - A new trend in the rational use of micelial wastes after antibiotics fermentation was elaborated. The modification of P. chrysogenum micelium by means of acid and alkaline solutions allows to use it as bone glue component. As a result physico mechanical characteristics of the mixture increase, while the ratio cost-price decreases. The obtained glue composition can be used in the production of glue paper tape, for paper and wood glue. PMID- 12728628 TI - [Comparative analysis of degradation of native avermectins and spinosyns by UV HPLC]. AB - Degradation rates and compositional changes in active ingredients of the two crop protection insecticides, Fitoverm and Spinosad, have been compared by using a reverse-phase HPLC with UV detection (250 nm). Decay of the major components of active ingredients: spinosyns A and D (Spinosad) and avermectins A1a, A2a, B1a and B2a (Fitoverm) was studied in the thin dry layer on the glass at sunlight at regular day/night changes of temperature. The following results were obtained: 1) 50% degradation time for spinosyns was about two times shorter than that for avermectins: at 40 degrees C day-time temperature it was 6 hours and 10 hours, respectively, while at 23 degrees C these times increased approx. ten-fold; 2) the initial composition of spinosyns was changed during degradation: ratio of spinosyns A/D was increased (i.e. D component degraded faster than the A one) and additionally 4-5 new spinosyns and/or their derivatives were formed; 3) rate of degradation of each avermectin was practically the same, i.e. percent composition of avermectins did not significantly alter; 4) retention times of avermectins B2a, A2a and A1a were similar to those of either initial spinosyns (A) or products of their decay. It is concluded that determination of spinosysn residues with the aid of UV-HPLC is a complex task since both initial spinosyns (A and D) and their conversion/decay products must be measured. The latter can be dominant residues and not always easy to identify. Analysis consider to be complicated when a sample contains residues of both spinosyns and avermectins. PMID- 12728629 TI - [Functional-metabolic activity of leukocytes and correction of it by lysozyme in acute cold stress]. AB - It was shown that immersion cooling caused deminished phagocytes number and and phagocytes index of polymorphonuclear leucocytes in blood, decreased NCT test data, decreased NADPH-oxidase activity, increased activity of Ca(2+)-ATPase in the cells. Lysozyme corrected the above mentioned parameters of polymorphonuclear leucocytes. Lysozyme activity was mediated by cytokins of glass-spleen cells. PMID- 12728630 TI - [Evaluation of levofloxacin antitubercular activity in vitro and in lung tissue culture]. AB - Levofloxacin in vitro demonstrated bactericidal effect against susceptible and multi-drug resistant Mycobacterium tuberculosis: range of MICs was 0.25-0.5 mcg/mL, MBC-0.5-1.0 mcg/mL. Postantibiotic effect after 24-hour exposition to bactericidal concentrations was 35-39 days. Levofloxacin possesed low toxicity when tested in mice lungs tissue culture--maximum safety concentration was 50 mcg/mL. Bactericidal effect of levofloxacin started three days after exposition and was maximal by 7 days of incubation: by this time mycobacterial microcolonies destruction started with detritus formation. It is emphasized that lung cells kept their viability completely. Combination of levofloxacin with isoniazide or pirazinamide resulted in strong synergistic effect obvious after 5 days of incubation, mycobacterial colonies destruction was registered by 7th day. Combination of levofloxacin with rifampicin resulted in antagonistic effect obvious by 7th day of the contact: the resulting effect was statistically significant and was manifested as microcolonies number and size enlargement when compared to data for single levofloxacin. PMID- 12728632 TI - [Pefloxacin (Abaktal)--efficacy in treating severe infections, optimization of treatment based on pharmacokinetic and pharmacodynamic parameters]. PMID- 12728631 TI - [Increase in efficacy of antibiotic therapy of anthrax under experimental conditions]. AB - The results of experimental therapy of antraxis infected mice with cefazoline (kefzol) and ampicillin incapsulated into liposomes are presented. Protective activity of the same free antibiotics combinated with amixine and leukinferone was evaluated also. Treatment with liposomal cefazoline enhanced mice survival upto 60 per cent, and life period upto 1.3 +/- 0.3 days. After liposomal ampicillin administration for 3 times the same indices were 60 per cent and 6.5 +/- 0.9 days, after 2 times administration--80 per cent and 14 +/- 1.8 days when compared to the groups of the animals treated with free antibiotics. It was shown that administration of ampicillin with amixine or with leukinferone provided enhanced mice survival upto 20 per cent, administration of cefazoline with leukinferone--upto 30 per cent. PMID- 12728633 TI - The high cost of merging with a religiously-controlled hospital. PMID- 12728634 TI - The impact of genetic studies of type 2 diabetes on clinical practice. PMID- 12728635 TI - Lessons learned from young-onset diabetes in China. AB - The prevalence of young-onset diabetes is rapidly rising in China. Young-onset diabetes is etiologically and phenotypically heterogeneous. Thirty percent to 50% of these patients have insulin secretory failure owing to autoimmune or monogenic or other yet to be identified forms of diabetes. Others have a strong family history of diabetes and exhibit features of the metabolic syndrome. Management of these young patients poses major diagnostic and therapeutic challenges, which require a multidisciplinary and holistic approach to ensure that these subjects are identified early and managed appropriately. Understanding the molecular basis of diabetes in these subjects may also eventually lead to improvement in diagnosis, classification, and treatment. PMID- 12728636 TI - Nutrition, growth, and body size in relation to insulin resistance and type 2 diabetes. AB - Nutritional and genetic factors interact in the etiology of type 2 diabetes. Undernutrition followed by overnutrition increases adiposity and the risk of diabetes. The thrifty hypotheses suggest that the nutritional challenges could have happened thousands of year ago (thrifty gene selection) or during one's intrauterine life (thrifty phenotype). Current strategies for the prevention of diabetes are related to avoiding overnutrition. PMID- 12728638 TI - Changing office practice and health care systems to facilitate diabetes self management. AB - Diabetes is a self-managed disease for which patients provide 99% of their own care. For patients to succeed as diabetes self-managers, they need office practices and health care systems that can prepare and support them in their diabetes self-management efforts over the long term. In order to provide effective diabetes education and ongoing support, office practices and health care systems will have to fundamentally redefine the roles of health professionals and patients with diabetes, and redesign practices and systems to allow for effective long-term self-management education and support. Although it is difficult for both people and systems to change, change is essential if we are going to provide self-management support for the majority of patients suffering from this serious chronic disease. PMID- 12728637 TI - The major diabetes prevention trials. AB - Type 2 diabetes (T2D) is a devastating disease with multiple complications affecting many tissues, most notably the cardiovascular system. To be able to prevent a chronic disease such as T2D, certain requirements have to be met. Knowledge about its natural history with a preclinical phase, modifiable risk factors, effective and simple screening tools to identify high-risk subjects, and effective intervention that is affordable and acceptable are necessary. In addition, the efficacy of the intervention has to be proven under a clinical trial setting. Several major lifestyle intervention trials have been successfully carried out, with consistent results: the risk of T2D in high-risk subjects can be halved, the effect of lifestyle changes is rapid, and benefits are similar in different ethnic groups. Thus, the prevention of T2D is possible in most high risk subjects, but how to achieve this at the population level remains a major challenge. PMID- 12728639 TI - Diabetes self-care: lessons from research on the family and broader contexts. AB - The foundation of diabetes management is the self-care behavior of the patient. All of the systems within which the person with diabetes interacts, as well as the media and broader social and cultural values, affect this self-care behavior. In this article I focus on recent research that has examined the link between relationships in the patient's intimate network (i.e., family and close friends) and in the patient's exchange network (i.e., patient-provider relationship, Internet support). The goal of this review is to identify relational targets associated with self-care behaviors that are potentially modifiable within the diabetes medical care setting. Evidence-based suggestions are made for points of intervention entry, and areas for future research are explored. PMID- 12728640 TI - Understanding and enhancing adherence in adults with diabetes. AB - This article reviews selected recent literature on adult diabetes patient centered adherence to diabetes care recommendations. We describe more fully a limited number of articles that have particular lessons for diabetes clinicians and researchers. Each article was reviewed for methodology, adherence measures, and types of outcome data collected. This process produced a few studies evaluating an intervention to promote adherence, subsequent targeted behavior changes, as well as health outcomes. Using a standard measure of diabetes self care adherence over multiple studies may be an important way to compare results across diverse interventions. Advances in research designs and measures show promising results for understanding and enhancing adherence in adults coping with diabetes. Much more behavioral research in diabetes is needed in order to significantly improve the health and quality of life in this population. PMID- 12728642 TI - Growing evidence for diabetes susceptibility genes from genome scan data. AB - Genome-wide scans for linkage have provided one of the dominant approaches adopted by researchers in their efforts to identify genes responsible for the inherited component of type 2 diabetes susceptibility. Around 20 genome scans have now been completed, in a wide variety of populations. Integration of data from these diverse scans has proven far from trivial, but the contours of genome wide linkage topography are steadily emerging from the fog of data. Identification of the calpain-10 gene as the probable basis for the chromosome 2q linkage seen in Mexican Americans has provided validation of this positional cloning approach. This report provides an update on recent type 2 diabetes genome scan data, focusing on several chromosomal regions where the evidence for linkage has considerably strengthened in the past year. The current and future value of genome-wide linkage information in the search for type 2 diabetes susceptibility effects is also discussed. PMID- 12728641 TI - The genetics of adiponectin. AB - Adiponectin encoded by the APMI gene is one of the adipocyte-expressed proteins that function in the homeostatic control of glucose, lipid, and energy metabolism. Its dysregulation has been suggested to be involved in disorders covering the metabolic X syndrome, such as insulin resistance, obesity, type 2 diabetes, and coronary artery disease. Recent data present evidence of a genetic modulation of the adiponectin level, and linkage of the 3q27 locus, where the APMI gene lies, with diabetes and features of the metabolic X syndrome playing a putative role of the APMI gene in this syndrome. In this article, we present an overview of the results available to date and discuss positive evidence for a role of genetic variants of the APMI gene and questions that genetic data raise. PMID- 12728645 TI - An Advisory Committee Statement (ACS). National Advisory Committee on Immunization (NACI). Statement on recommended use of pneumococcal conjugate vaccine. PMID- 12728644 TI - Quantitative trait linkage studies of diabetes-related traits. AB - Genetic linkage methods for diseases with complex inheritance are based on assessment of allele sharing between affected relative pairs, but such methods have low power to detect genes with moderate effects. This may explain the difficulty in replication for many of the putative loci for type 2 diabetes. To enhance power to detect diabetes-susceptibility genes, some investigators have performed quantitative-trait linkage studies for diabetes-related traits, including measures of glycemia, insulin sensitivity, insulin secretion, obesity, lipidemia, and blood pressure. These linkage studies have not provided stronger or more consistent evidence for linkage than studies of diabetes affection status, but have identified several loci that may play an important role in the physiologic processes related to the development of type 2 diabetes. PMID- 12728646 TI - An advisory committee statement (ACS). National Advisory Committee on Immunization (NACI). Statement on smallpox vaccination. PMID- 12728647 TI - HealthSouth's go-to guy. PMID- 12728643 TI - Lessons for human diabetes from experimental mouse models. AB - A precise knowledge of the defects underlying type 1 and type 2 diabetes is essential for designing appropriate therapeutic strategies. Because experiments in humans are limited, naturally occurring, and especially genetically engineered rodent models, have revolutionized research in diabetes. We review some of the models created recently and discuss their impact on human diabetes. PMID- 12728648 TI - Masks can't stop this virus. PMID- 12728649 TI - Flying scared. Seven ways to make air travel safer. PMID- 12728650 TI - Altria's perfect storm. Hit by cut-rate competitors, taxes, and most of all, litigation, the company that owns Philip Morris faces its worst crisis in years. PMID- 12728651 TI - Spinning the wheel on ImClone. PMID- 12728652 TI - I'm not paranoid, I just live here. PMID- 12728653 TI - [Tuberculosis at the beginning of the 21st century]. PMID- 12728654 TI - [Mortality and tuberculosis: analysis of multiple causes in the Community of Madrid (1991-1998)]. AB - BACKGROUND: Mortality from tuberculosis (TBC) is of characteristics such as to be studied with a multiple cause focus, as this disease is not generally fatal in itself, but is however linked to many deaths. This study is aimed at describing TBC as a cause of death in the Autonomous Community of Madrid throughout the 1991 1998 period. METHOD: A descriptive study of deaths in the Autonomous Community of Madrid (1991-1998) entailing TBC as the main cause or as a diagnosis mentioned in the Death Statistics Bulletin (DSB). The data was taken from the Autonomous Community of Madrid Death Registry. Variables analyzed: sex, age, year deceased, main cause and other diagnoses mentioned in the DSB. Crude annual rates, standardized rates by age and specific rates by age and sex were calculated. A comparison was made as to the average number of diagnoses per death. RESULTS: A total of 1,206 deaths "with TBC", 915 males and 291 females, were recorded. The main cause was TBC in 566 of these cases, HIV/AIDS in 442 and other diseases in 198 cases. The "with TBC" death rates showed peak values in 1995 among both males and females. There were an average of 3.4 diagnoses per death, this average having undergone an increase throughout the period under study (p < 0.05) CONCLUSIONS: With a multi-cause focus, TBC doubles in death rate statistics. Its presence is associated with other diseases, especially with HIV/AIDS for the period analyzed. PMID- 12728655 TI - [Occupational exposure to multiresistant Mycobacterium bovis in a hospital in Saragossa, Spain]. AB - BACKGROUND: Those working in a hospital environment are exposed to different occupational risks, although more specifically biological ones. One of the major risks is that of exposure by airborne transmission, more specifically, tuberculosis. This study is aimed at providing a description and analysis of the results of the implementation of an evaluation and surveillance protocol following occupational exposure to Multiresistant mycobacterium bovis (MRMb). METHOD: A male patient was diagnosed with MRMb infection at the Miguel Servet Hospital in Zaragoza in 1999 following ten days without respiratory isolation. During this period, he came into contact with 167 employees from different hospital departments. A surveillance and contact control protocol was prepared based on: completing a survey and undergoing an initial Mantoux (if the employee had previously tested negative for tuberculin), followed three months later by a chest X-ray and then a two-year clinical follow-up (check-ups every three months) for those having tested positive for tuberculin and no administering of chemoprophylaxis even though signs of infection were to have been found. RESULTS: Information was gathered on 160 employees (96%). A total of 94 employees (59%) had previously undergone a Mantoux, seven (7) having had tuberculosis. It was necessary for a follow-up to be conducted on sixty-one (61) employees who tested positive (29 previously positive and 32 detected in the initial Mantoux). No employee who had tested negative on an initial Mantoux tested positive on repeated testing three months later nor showed any symptoms indicative of transmission during the follow-up period. Some variables, such as age or working in the Infectious Disease Unit were related, on a statistically significant basis, to follow-up being required. CONCLUSIONS: The risk of occupational transmission following contact with MRMb might be similar to M. Tuberculosis, although further experience would be required in order to confirm this fact. Early diagnosis and availing of a protocol for implementing measures aimed at preventing and controlling this type of occupational exposure are of importance. PMID- 12728656 TI - [Epidemiological surveillance of pulmonary tuberculosis treated at the specialized care level based on 2 data sources, Valladolid; Spain]. AB - BACKGROUND: Pulmonary tuberculosis is still more frequent that it should be in Spain given the degree of Spain's social and healthcare-related development. Apart from some individual studies, such as the Multicenter Tuberculosis Research Project, the incidence of tuberculosis is known by way of the Compulsory Notifiable Disease System, in which some degree of under-notification has been detected. The question has been raised as to whether this data can be improved through the additional use of another registry, specifically the Minimum Basic Data Set (MBDS). METHODS: This is a retrospective study referring back to the 1996-2000 period conducted on the population of a healthcare district totaling 220,572 inhabitants. The data from the Compulsory Notifiable Disease registry was used to the specialized care level, and that of the MBDS registry for the diagnosis of pulmonary tuberculosis. The incidence rates were calculated for each source by the capture-recapture method. An analysis was made of epidemiological characteristics such as age, gender, place of residence, bacillus in sputum and treatment data on the hospitalized cases, such as average length of stay, type of admission, type of release, clinical department, HIV co-morbility. RESULTS: The mean annual incidence recorded at the specialized care level was 16.6 cases/100,000 inhabitants in the Compulsory Notifiable Disease registry; 20.4 cases/100,000 inhabitants in the MBDS registry, and 23.1 cases/100,000 inhabitants combining both of these two sources. The incidence estimated using the capture-recapture method was that of 24.4 cases/100,000 inhabitants (IC95%: 23.5-25.3). CONCLUSIONS: Any information system which provides reliable data serves to improve epidemiological surveillance even though it may have been designed for a different purpose. It is all a matter of knowing the limitations and unique aspects thereof. The MBDS provides information of epidemiological interest which is not included in the Compulsory Notifiable Disease reports. Using the capture-recapture method is one alternative for estimating truer pulmonary tuberculosis rates. PMID- 12728657 TI - [Incidence of tuberculosis at the local level: Marianao Municipality, Havana City, Cuba (1990-2000)]. AB - BACKGROUND: For the elimination of tuberculosis (TB) is necessary the surveillance of the incidence rate at the local level, as this is the first level of access to the health service. This study is aimed at providing a description of the spread and trend of the tuberculosis incidence rates in Marianao Municipality, Habana City by healthcare areas and age groups throughout the 1990 2000 period. METHODS: The time series for the 1990-1994 and 1995-2000 incidence rates were analyzed employing a two parameter exponential smoothing method. The total percentage and annual average of variation was estimated. The healthcare areas were stratified. RESULTS: In 1990-1994, the rate rose from 11.1 to 25.8 per 105 inhabitants (33% of increase by year), having been most noticeable in Carlos J. Finlay and Portuondo health areas (respectively annual average 112.7% y 36.2% increases). Taking into account that the control program was strengthened in 1995, the rates dropped by 47.6% in this municipality during the 1995-2000 period. The Finlay and Portuondo areas respectively lowered their rates by 79.9% y 39.7% while the Gonzalez Coro area showed a 58.3% rise and 27 de Noviembre area had no major change. The age and above 60 group had the higher percentage of cases, recording 49.8% of the total and only one case among children under age 15 for the period. During this same period, only six cases of tuberculosis/human inmunodeficiency virus were diagnosed. The 27 de Noviembre area was the only one found to be at the unsatisfactory level according to the stratification. CONCLUSIONS: An initially upward trend, followed by a downward trend in tuberculosis was found to exist in the Marianao Municipality throughout the period studied. This disease was predominant among older adults. PMID- 12728658 TI - [Study on tuberculosis in a Seville Health Care District. Current situation and control improvement alternatives]. AB - BACKGROUND: In our setting (District, Seville, Andalusia, Spain....) it is not rare to find annual incidence rates for TBC of more than 30 new cases per 100,000 inhabitants. It reveals that this problem is nowhere near being eradicated. This study is aimed at describing the clinical and public health patterns of tuberculosis cases within the "South-East Seville" Healthcare District. METHODS: Descriptive study of the reported cases (personal traits, location, time, type of disease, risk factors) throughout the 1992-2000 period in the "South-East Seville" Healthcare District, located in the city of Seville and including some rural centers of population. Percentage distribution of cases for the different studied factors and the incidence rates for group of interest (sex, age groups, geographic areas) are provided. RESULTS: A mean annual incidence rate of 19.4 cases/100,000 inhabitants has been estimated. Notable differences in incidence depending on group of sex (RR = 2.1), age (annual incidence of more than 24 cases/100,000 inhabitants among children age 0-4 and adults age 25-39) and geographical area were found. The frequency of relapses and repeated treatments, as well as shortcomings with regard to conducting and reporting the studies on family members and contacts were noted. CONCLUSIONS: The situation analysis in a Healthcare District such as the one described here (currently approx. 610,000 inhabitants) throughout a nine-year follow-up period -entailing 1,065 reported cases- may well provide us some orientation about the situation in our environment. It also gives us the possibility to compare it with some other studies. A downward trend in the incidence rate has been noted as of 1997. Some organizational measures to be taken into account for controlling this infection are provided. PMID- 12728659 TI - [High prevalence of malnutrition among the indigenous early childhood population in Mexico. National Nutrition Survey 1999]. AB - BACKGROUND: Malnutrition among the indigenous early childhood population is still currently a serious public health problem, and given that no specific studies affording the possibility of knowing the nutritional condition of this population, the question was posed of identifying the prevalence of malnutrition among the indigenous population of Mexico by means of the 1999 National Nutrition Survey. METHODS: A rural sample was selected from the Northern, Central and Southern regions where 70% or more of the population speak an indigenous language. The weight/age, height/age and weight/height Score Z desviations was calculated for 3,236 preschoolers and 4,899 school-age children. RESULTS: Nationwide and by regions, there were no difference by age group regarding the prevalences of the three indicators. By regions, for preschoolers, the South showed the highest prevalences of underheight (69.8%) and underweight (49.8%) than the North (respectively 36.1% and 22.6%), entailing statistically significant differences (p = 0.0002 and p = 0.04). For overweight and obesity, the North showed a 14.2% prevalence, and the South 5.6% (p < 0.05) for the same age group. CONCLUSIONS: The nutritional condition of the children studied reveals a geographical polarization, the greatest prevalences related to underheight and underweight children being found in southern Mexico, whilst those due to overalimentation were located in the North, this phenomenon possibly being due to lifestyles and availability of food differing from one region to the other. PMID- 12728660 TI - [Health-related needs of the displaced population due to armed conflict in Bogota]. AB - BACKGROUND: The displacement of populations as the result of an armed conflict generally entails a worsening of the living and health conditions of those undergoing such a displacement. This paper is aimed at analyzing the health related needs perceived by men and women displaced by the armed conflict and their main strategies to address those needs. METHODS: A qualitative study was carried out by means of semi-structured individual interviews to 31 displaced men and women in the transition stage. A narrative analysis of the contents was conducted, segmenting the information by age and sex. The area under study was made up of five localities in the city of Bogota. RESULTS: The negative effects on their mental health and psychosocial stability, access to food and, to a lesser degree, gastrointestinal and respiratory disorders, are the main health problems reported by both groups of informants. The difficulty of accessing health care services comes up as an added problem. The precarious economic situation underlies the health care-related needs and problems. To solve their health problems, in addition to the health care services, they employ other strategies within their reach. Expressed needs are coherent with the problems perceived. Some differences between women and men and age groups were observed concerning the definition of the problems and employed strategies. CONCLUSION: In the health field, actions are required in order to improve their access to services in addition to specific strategies for the psychosocial rehabilitation of the displaced population which take into account the differences existing within this group. PMID- 12728661 TI - [Pattern of use of the prenatal, childbirth and puerperium care services at a Mexican Social Security Institution]. AB - BACKGROUND: Maternal health is a priority in the Mexican health system, the proper planning and organization of the resources, as well as the use of the maternity care services suited to the needs of pregnant women therefore being of importance. This study is aimed at determining the pattern of use of the prenatal, childbirth and puerperium care services at a Mexican social insurance institution. METHODS: A descriptive study was conducted of 403 women involved in an obstetrical medical event, with the exception of those whose clinical file was not located or who were treated at a subdistrict hospital providing a low degree of coverage of obstetrical events and in a third-level hospital. An analysis was made as to the type and frequency of use of the healthcare services in the prenatal stage, the childbirth care provided up to the immediate puerperium and the advanced puerperium stage. The information was taken from the clinical file, both at the second-level hospital as well as in family medicine and, if required, at the third-level hospital. RESULTS: 90.8% were women whose pregnancies ended in vaginal birth or cesarean section, their prenatal care visits averaging 6.2 +/- 4.1. Prenatal care was started during the first three months by 48.6%, whilst 27.3% came in for visits 7 or more times. A total 22.4% showed both characteristics. The average number of days of hospitalization was 1.8 +/- 0.9 days. During the puerperium, office visits averaged 1.2 +/- 0.4. CONCLUSIONS: Less than one fourth of the population studied kept the prenatal care appointments as should have been, mention being made of areas of opportunity in childbirth care. PMID- 12728662 TI - [Health education in the Spanish school manuals]. AB - BACKGROUND: The school manual or textbook is the educational tool most used by the teaching staff. This study is aimed as ascertaining how Education for Health is dealt with in Primary and Secondary school textbooks, which is of utmost importance as regards finding out how they are implemented in the classroom, as well as setting out some points for thought for the authors and editors of these texts and for the teaching staff and healthcare personnel providing assistance to the schools. METHOD: A total of 297 textbooks were analyzed, 123 of which were used at the Primary level, 149 at the Compulsory Secondary level and 25 at the Higher Secondary level. This study deals with five aspects: 1) Presence or absence of Education for Health 2) Health-related topics included 3) Degree of health covered 4) Included in the subject or dealt with on a transversal basis and 5) Methodological aspects. A number of criteria for analysis have been defined for each one of these aspects which will make it possible to consider the degree to which they are covered. RESULTS: Sixty-three percent of the books analyzed included topics related to Education for Health. The topics dealt with most were related to matters of hygiene (39.5%), followed by eating (26.5%). The health-related concept most often dealt with is that of being disease-free (36.9%) and a state of well-being (20%). Somewhat over half of the textbooks dealing with health provide learning activities, although only in certain isolated cases (57.7%). The methodology most often used is informative explanation (32.4%) without dealing with the procedure-related contents or attitude-related criteria. CONCLUSIONS: The current school manuals are not sufficient either as a point of reference or as an adequate resource in order for the teaching staff to provide Education for Health in the classroom, as they do not place sufficient emphasis thereon despite its being considered a transversal subject, and when they indeed do so, the way in which they are oriented does not set attitudes or behaviors into motion. PMID- 12728664 TI - [Unexpected properties of angiotensin mediated by the AT2 receptor: possible therapeutic implications]. AB - The renin-angiotensin-aldosterone system plays an important role in the regulation of electrolyte balance, body fluid volume and blood pressure. As yet, angiotensin II has been ascribed actions mediated by first type of receptors (AT1) such as increased blood pressure, antinatriuretic effect, cell proliferation. Since several years studies have been conducted on the role of second type receptor (AT2) through which angiotensin manifests its effects opposing those resulting from stimulation of type 1 receptor. They include: release of bradykinin and nitric oxide, vasodilation, natriuretic and antiproliferative effects. Blockade of the function of this receptor causes excessive reaction induced by action exerted on AT1 receptor. PMID- 12728663 TI - [Outbreak of gastroenteritis caused by Norwalk virus at a senior citizens assisted living facility in Granada, Spain]. AB - BACKGROUND: The outbreaks of acute gastroenteritis caused by a virus are currently one of the health problems having the greatest impact on those living in senior citizen facilities. The objective of this study is that of investigating the causes of the start and contagion of an outbreak of acute gastroenteritis caused by a Norwalk-Like virus. METHOD: At a senior citizen living facility with 141 residents and 71 employees exposed, the relations between the disease in question and different independent variables: personal traits, features of location, date of onset of symptoms, clinical pattern of the disease and causal factors (food and drinking water intake). A descriptive study, a contingency table and a hypothesis ?2 test, design of cases and controls with the calculation of the unprocessed and subsequently adjusted Odds Ratio (IC 95%) were carried out using logic regression. RESULTS: No significant differences were found in the personal trait and features of location variables. The attack rate among the residents was 30.1%, being 21.12% among the employees. In the adjusted Odds Ratio calculation, solely the whipped cream mousse showed causal relationship values (adjusted Odds Ratio = 4.66; IC-95% 1.15-18.91; p = 0.031). Ten patient stool samples were sent for analysis, no disease-causing bacteria having been detected, Norwalk-Like virus however having been isolated in three samples. CONCLUSIONS: The whipped cream mousse having been found to possibly have been responsible for the outbreak is a first involving Norwalk virus, but this result is subject to limitations, such as the possible bias of classification due to failing to recall having eaten foods. Even so, this is an element to be taken into account in the control of acute viral gastroenteritis. PMID- 12728665 TI - [ACE gene polymorphism and renal scarring in children with urinary tract infection and vesicoureteric reflux: preliminary results]. AB - To investigate angiotensin-converting enzyme (ACE) gene deletion/insertion (D/I) polymorphism as a risk factor of renal scarring in children, we determined ACE genotypes in 63 paediatric patients with primary vesicoureteric reflux (VUR) grade I-V and recurrent urinary tract infections (UTI). Renal cortical scars were evaluated applying Technetium-99-m-unithiol SPECT method in 21 children. No statistically significant differences were found in allele and genotype distribution of the D/I polymorphism within ACE gene between children with VUR/UTI and healthy Caucasians (n = 806, after meta-analysis of literature data) (D allele frequencies--47% vs 54%, respectively). It suggests that ACE gene polymorphism is not associated with primary VUR. Renal scarring was revealed in 7 out of 21 children and there was also no statistically significant difference in D/I distribution between scar (n = 7) and non-scar (n = 14) subgroups of patients and controls. However, higher incidence of D allele (64%) in children with renal parenchymal damage could suggest the hypothesis that this genetic factor plays an important role in renal scarring. Therefore, to verify this assumption further studies on a larger group of patients are needed. PMID- 12728666 TI - [Increased thrombin-genesis in cyclosporine A-treated idiopathic nephrotic syndrome in children]. AB - Cyclosporine A (CsA) has been accepted as one of the most efficient therapies of idiopathic nephrotic syndrome (INS) in children. Despite its beneficial effect on clinical course of the disease, its use has been associated with a number of side effects. This prompted us to study the influence of cyclosporine A on the coagulation cascade in nephrotic children. We examined thrombinogenesis in 16 children in remission of steroid-dependent idiopathic nephrotic syndrome treated with cyclosporine A. The concentrations of F1 + 2 prothrombin fragments and thrombin-antithrombin complexes were used as markers of coagulation cascade activation. The results were compared between 18 children with INS relapse who had responded to 8-week glucocorticoid treatment (not treated with cyclosporine A) and 20 healthy subjects. We found an increased concentration of F1 + 2 prothrombin fragments in children treated with CsA, while in children after 8 weeks of glicocorticoid therapy the concentration of this marker was comparable to that in the controls. Since we observed similar biochemical disturbances in both INS groups, we suggested that cyclosporine A was able to stimulate coagulation that might lead to an increased risk of thomboembolic events in children with clinical remission of INS. PMID- 12728667 TI - [Analysis of renal replacement therapy in patients with renal failure]. AB - The article presents the 10-years experience of renal replacement therapy in a single centre. A total of 158 patients were treated in this period. 77 patients (47 F, 30 M, mean age 18.7 +/- 12 yrs) were treated due to chronic renal failure and 81 (35 F, 46 M, mean age 2.1 +/- 1.5 yrs) due to acute failure. 48 (62%) were treated by haemodialysis, 24 (31%) by peritoneal dialysis and 5 (7%) by both methods. Due to the shortage of dialysis units both children and adults were qualified to the dialysis therapy in our centre. Mean age of haemodialysis patients was 22 yrs and of those treated by peritoneal dialysis 13.5 yrs. 7 patients died (9%) and 30 (39%) were transplanted. 15 (19%) were transferred to other centres. The overall mortality was lower than reported by other authors. 4% of patients were tested HBV positive, 13% HCV positive and 9% both HBV and HCV positive and this rates are lower than the average rate in chronic dialysis patients in Poland. Our experience allows us to conclude that adult patients may be successfully treated in paediatric dialysis centres. PMID- 12728668 TI - [Evaluation of health-related quality of life in dialysis patients. Personal experience using questionnaire SF-36]. AB - INTRODUCTION: Patients' perception of health is an important outcome measure in the assessment of influence of chronic disease and received treatment. The purpose of this study was: 1) to investigate the relation between selected demographic and clinical characteristics and Health-related Quality of Life (HRQoL) scores in patients with end-stage renal disease (ESRD) who receive dialysis, 2) to compare HRQoL in dialysis patients with their peers from the control group, 3) to evaluate the usefulness of SF-36 questionnaire. MATERIAL AND METHODS: We evaluated Health-Realated Quality of Life (HRQoL) of dialysis patients by the Polish version of the Short Form 36 Items Health Survey (SF-36). The SF-36 consists of eight scales: physical functioning (PF), social functioning (SF), role limitation attributable to physical problems (RP), role limitation attributable to emotional problems (RE), mental health (MH), vitality (VT), bodily pain (BP) and general health perception (GH). This study included 97 patients (44 men and 53 women), aged 28-86 years (mean age: 57.2) treated with HD (n = 77) and CAPD (n = 20) for 3-245 months (mean: 61 months). We compared the results with the data obtained from 217 healthy control subjects (105 men and 112 women), aged 20-92 years (mean age 51.1). The patients were divided into four age groups and compared with the appropriate groups of controls. RESULTS: The perception of health of dialysis patients was worse than that of controls. We have found, that in the group aged over 65 years patients' scores were quite close to those of the control population. Our results show the following patient connected factors to be independently associated with quality of life (QoL): age, sex, occupation, level of education, family situation and comorbidities. On average, females reported lower scores; the impact of ageing was more evident in physical scales. CONCLUSIONS: 1) age, sex, occupation, level of education, family situation as well as comorbidities are independent factors of HRQoL, 2) subjective QoL of elderly patients seems acceptable in comparison with healthier peers, 3) the SF-36 questionnaire is applicable in dialysis patients and SF-36 scores are related to important clinical aspects. PMID- 12728669 TI - [Use of minimally invasive direct coronary bypass (MIDCAB) in reoperative coronary surgery]. AB - The authors presented the method and early results of minimally invasive direct coronary arteries reoperation in 5 patients, approximately 7, 5 years after the first CABG operation. In all patients venous grafts were occluded. there was a different approach used in every patient: small left or right anterior thoracotomy through IV or V intercostal space, laparotomy with partial low sternotomy or simultaneous combined approach through thoracotomy and sternotomy/laparotomy. The following arteries were grafted: left anterior descending, right posterior descending, LAD with right coronary artery and LAD with marginal branch. All patients were extubated in the operating room. There were no serious perioperative complications observed. Patients were transferred to cardiology department on the 5th-7th postoperative day. PMID- 12728670 TI - [Secretion of interleukin 2, 6 and tumor necrosis factor by peripheral blood mononuclear cells in patients with uveitis induced by phytohemagglutinin or lipopolysaccharide]. AB - The aim of the study was to determine the concentration of selected cytokines secreted by peripheral blood mononuclear cells (PBMC) of patients with uveitis. We determined the spontaneous secretion and PHA- or LPS-induced production of IL 2, IL-6 and TNF in supernatants. The concentration of examined cytokines in patients with uveitis in comparison with healthy controls was not statistically significantly higher. PMID- 12728671 TI - [The evaluation of natural anticoagulants in systemic lupus erythematosus in children]. AB - Thromboembolic complications in adults with systemic lupus erythematosus (SLE) are described in literature. We intended to investigate the activity of natural anticoagulants, such as C protein. S protein and antithrombin III (AT III) in children with SLE to obtain data concerning activity of the disease and thrombotic complications. The study population consisted of 36 children with SLE and of 51 healthy children serving as a control group. The results showed a significant decrease of S protein activity, with particularly low levels in the group of patients with the presence of anticardiolipin antibodies (aCL), and decrease of C protein activity in patients with higher activity of the disease. They appear to prove the involvement of these inhibitors in inflammation and, on the other hand, the activation of intravascular coagulation, a risk factor of thrombosis. PMID- 12728672 TI - [Cyclosporine in the treatment of atypical celiac sprue. A case report]. AB - We present a 45-year-old woman with atypical celiac sprue who was unresponsive to gluten-free diet. Cyclosporine therapy was applied (5 mg/kg body weight daily for 2 months). The patient demonstrated a remarkable clinical and histological response to this drug. PMID- 12728673 TI - [Features of generalized infections masking basic disease: non-Hodgkin's large cell anaplastic lymphoma and hemophagocytic syndrome shown in two patients]. AB - The authors describe diagnostic problems encountered while treating two children finally diagnosed to have anaplastic lymphoma in the first case and infection associated haemophagocytic syndrome (IAHS) in the second. Both diseases represented secondary immunodeficiencies masked by local and systemic severe infections. PMID- 12728674 TI - [Application of pharmacotherapy with classic neuroleptics: phenothiazine derivatives in schizophrenia]. AB - Tricyclic psychotropic drugs (TDP) are used in the therapy of both schizophrenia and affective disturbances, especially depressive ones. They are a large group of compounds with a broadband spectrum, especially as an antipsychotic and antidepressant drugs. Phenothiazine derivatives distinguish themselves with a structure of triple cycles arrangement, composed of electrons such as a sulphur and nitrogen atoms. Among patients with schizophrenia there are many groups with different psychopathological symptoms, course of disease and neuroleptics drugs response. Affective disturbances occur particularly often in the course of schizophrenia. A higher incidence of depression in the course of schizophrenia has been observed during treatment with neuroleptics, especially in the late phase of treatment, particularly after using strong neuroleptics. PMID- 12728675 TI - [Harmonic imaging in ultrasonography: basic concepts and applications]. AB - Current ultrasound techniques using harmonic imaging are presented. Narrow-band harmonic imaging, pulse inversion and coded harmonic imaging techniques are discussed. Basic information about contrast-enhanced harmonic imaging techniques (linear and harmonic scatter, stimulated emission, Flash Echo and subharmonic imaging) is given. Principal advantages of harmonic ultrasonography and clinical indications are discussed. PMID- 12728676 TI - [Modern possibilities of bioterrorism diagnosis]. AB - One of the most important components of early detecting and response to biological attack are microbiological diagnostic methods. In order to provide the effectiveness of diagnosis continuous training in detection of potentially dangerous bioterrorist agents is needed. Basic information about selected microbiological agents most dangerous for public health, belonging to A and B CDC categories are presented in the article. The most potent biological weapon mentioned in the last decade is anthrax. Other potential bioterrorist agents discussed in this article are Francisella tularensis, Yersinia pestis, Brucella species and Coxiella burnetii. PMID- 12728677 TI - [Viral hemorrhagic fevers as a biological weapon]. AB - Viral haemorrhagic fevers are zoonoses caused by a group of phylogenetically diverse RNA-viruses, capable of causing serious haemorrhagic complications in humans. The West-African Ebola and Marburg viruses pose the most significant threat because of their easy spreading through direct contact with the ill person and high death rate reaching 90%. They are considered among the most dangerous agents possibly used in bioterrorist attack and have been studied as a part of the Soviet biological weapons programme. The first symptoms of the Ebola haemorrhagic fever appear 4 to 16 days after the infection and are rather unspecific (fever, flu-like and gastrointestinal symptoms, cough, sore throat, conjunctivitis). Within a few days the disease leads to weight loss, haemorrhagic complications and circulatory insufficiency. The infection may be transmitted through direct contact with the patient, his/her body fluids and cadavers; droplet transmission is much less likely. There is no specific prophylaxis nor treatment; still, isolation of patients and use of personal protection means by persons providing care to patients seem efficient in stopping the infection. The knowledge of the biology and epidemiology of Filoviridae is still limited, which makes the results of bioterrorist attack using these pathogens hard to predict. PMID- 12728678 TI - [Smallpox virus as biological weapon]. AB - Smallpox, because of its high case-fatality rate, easy transmission from human to human, lack of specific treatment represents nowadays one of the main threats in bioterrorist attacks. Over the centuries, naturally occurring smallpox with its case-fatality over 30 percent and its ability to spread in any climate and season has been treated as the most dangerous infectious disease. But it is now, 25 years after the last documented case of smallpox and cessation of routine vaccination in present mobile and susceptible population, smallpox virus spread might be rapid and devastating. PMID- 12728679 TI - [Botulinum exotoxin as an instrument for a bioterrorism attack]. AB - In this paper epidemiological and clinical aspects of intoxication with Clostridium botulinum are discussed. The attention is paid to the possibilities of using this toxin as a bioterrorist weapon. PMID- 12728680 TI - [Anthrax as a pathogen for a biological weapon]. AB - The authors presented an outline of modern knowledge about anthrax. Some aspects of the use of Bacillus anthracis as a biological terrorist agent are also described. PMID- 12728681 TI - [New views on systemic mastocytosis]. AB - Systemic mastocytosis is an uncommon disease characterised by an abnormal increase of mast cell count. The observed symptoms result from bone marrow, gastrointestinal, hepatic, splenic and skin infiltration by mast cells and from release of mast cell active mediators. The aim of our study was to discuss the classification, clinical features and treatment opportunities of systemic mastocytosis, based on review of current literature. PMID- 12728682 TI - [The role of hormone replacement therapy in the treatment of women with complicated chronic renal failure]. AB - Patients with chronic renal failure (CRF) show an increased mortality and morbidity from cardiovascular disease. Women with CRF are also at high risk of accelerated bone loss and fracture because of complications such as renal osteodystrophy and many endocrine disturbances leading to premature menopause and estrogen deficiency. Hormone replacement therapy (HRT) has been known to have beneficial effects on various atherosclerotic parameters in population of postmenopausal women and its role in primary prevention of coronary artery disease has been proved in many studies. In general population estrogen replacement therapy has also been used as an effective form of treatment in women suffering from bone loss as well as a form of prevention of osteoporosis. Until now, the role of estrogen replacement therapy in the treatment of complications of end-stage renal disease in female patients has not been fully elucidated and the use of hormone replacement therapy needs to be carefully analysed in each individual case taking into account all risks and benefits. PMID- 12728683 TI - [Hypertensive nephropathy: pathogenesis, diagnosis and treatment]. AB - Chronic hypertension leads to renal damage known as hypertensive nephropathy or hypertensive nephrosclerosis. Presently, this renal pathology seems to be one of the most frequent causes of end-stage renal disease requiring chronic dialysis. Unfortunately there are many problems with clinical diagnosis of hypertensive nephropathy which cause overestimation of its prevalence. Renal biopsy and morphological examination which is the gold standard of diagnosis is rarely performed in such patients. Histological lesions in hypertensive nephropathy are well recognised: myointimal hyperplasia of arterioles, hyaline arteriosclerosis, wrinkling of basement membrane, collapse of the glomerular tuft, glomerulosclerosis and tubulointerstitial involvement. These changes are believed to result from ischaemia and hyperfiltration. Genetic susceptibility and enhanced apoptosis could be involved in this process as well. One could hope that adequate treatment of hypertension according to international standards would allow to reduce the still increasing number of patients with hypertensive nephropathy. Pathogenesis, epidemiology, and clinical management of hypertensive nephropathy are currently discussed in this brief review. PMID- 12728684 TI - [Contemporary opinions on liver fibrosis pathogenesis]. AB - The author presents the selected aspects of liver fibrosis pathogenesis with special regards to the mechanism of hepatic stellate cells activation. PMID- 12728686 TI - [Migraines and headaches]. PMID- 12728685 TI - [Nutrition in health establishments. An innovative national program]. PMID- 12728687 TI - [Headache Emergency Center, Paris, Lariboisiere Hospital]. PMID- 12728688 TI - [Nursing care organization at the Headache Emergency Center]. PMID- 12728689 TI - [Nursing aides' role]. PMID- 12728690 TI - [Permanent training. For a more equitable access]. PMID- 12728691 TI - [Update: hospital strikes]. PMID- 12728692 TI - [Methadone]. PMID- 12728693 TI - [Implantable automatic defibrillator: the French delay]. PMID- 12728694 TI - [The cancer patient at home: coordination of care]. PMID- 12728695 TI - [Peer communication skills and social behavioral characteristics of preschool children]. AB - The purpose of this study was to investigate the relationship between peer communication skills and social behavioral characteristics of preschool children. Data for 42 preschool children, five to nine years old, were analyzed in the study. Teachers were asked to rate the children on a number of social behavior measures, and classify them into competent, aggressive and withdrawn groups. It was shown that children of the withdrawn group accepted more requests from others than those of the other groups. Generally, in terms of feedback they gave, the withdrawn were not much different from the competent, but forms of feedback were more often nonverbal. As for the aggressive, feedback was more verbal than the withdrawn when another directly spoke to them, but tended to be equally nonverbal when it was not clear whom other were speaking to. PMID- 12728696 TI - [The effects of hypnosis on emotional responses of depressed students in frustrating situations]. AB - The present study examined the effects of hypnosis on aggression and depression in depressed undergraduate students. Six frustrating situations were presented to 13 mildly depressed subjects as well as to 13 non-depressed subjects. All subjects were studied both in light trance and in the waking state. Their emotions were measured by numeric rating scales and open-ended questions. On scores using with numeric rating scales, depressed subjects were less depressive under trance as compared to the waking state. Responses to open-ended questions, which were scored by four raters, indicated that depressed subjects were less aggressive during trance than during the waking state, while non-depressed subjects were more aggressive under trance. Furthermore, the depression score was lower during trance than during the waking state. These findings suggested that depressed subjects were not repressive, while non-depressed subjects were repressive and exhibited controlled anger in the waking state. PMID- 12728697 TI - [The properties of visual short-term memory on change detection task]. AB - Visual short-term memory (VSTM) has been investigated with a change detection task. Recent studies suggested that there might be some representations in VSTM even when a change was not detected. However, this is discrepant with the previous studies that estimated the representation by change detection. In this study, we investigated the properties of the representation to be retained between two stimuli in a change detection task combined with the probe method so as to explore what causes the discrepancy. The interval between the test and the comparison stimuli and the timing of a positional cue at the location of change were manipulated. The results of three experiments suggested that, before the comparison stimulus presentation, the representations in VSTM were retained more than representations estimated by a normal change detection task, that they decayed with time, and that their availability decreased when the representations of the comparison stimulus were formed. From these results, we discussed a model of VSTM with attention. PMID- 12728698 TI - [Learned helplessness, generalized self-efficacy, and immune function]. AB - Generalized self-efficacy is considered one of important personality traits that determine psychological and physiological stress responses. The present study examined the interaction effects of generalized self-efficacy and controllability of acute stress on salivary secretory immunoglobulin A (s-IgA), task performance, and psychological stress responses in a typical learned helplessness paradigm. Twenty low and 19 high self-efficacy undergraduate women performed two response selection tasks one after another. In the first task, they were exposed to controllable or uncontrollable aversive noise. The second task was identical for all, but perceived controllability was higher for the high self-efficacy group than the low. Performance under uncontrollable condition was lower than controllable condition. The interaction of self-efficacy and controllability was observed only on the s-IgA variable; increase of secretion of s-IgA secretion under stressor uncontrollability was more prominent in the low self-efficacy group than the high. These results suggested that generalized self-efficacy was a moderator of the stressor controllability effect on secretory immunity. PMID- 12728700 TI - [Ongoing group interaction, ingroup favoritism, and reward allocation]. AB - Two alternative explanations for ingroup favoritism in the minimal group situation have been proposed: social identity and expectations of bounded generalized reciprocity. In this study, predictions derived from the two were examined for ingroup favoritism in a realistic group salutation with ongoing interactions. Results of the experiment, a replication of Karp, Jin, Yamagishi, and Shinotsuka (1993) which used groups with ongoing interactions rather than minimal groups, were inconsistent with either of the two explanations. (1) Ingroup favoritism emerged even in the condition under which the nature of fate control was unilateral. The finding was in sharp contrast to results of previous minimal groups experiments; the studies found ingroup favoritism only when fate control was mutual. (2) Analysis of post-experimental questionnaire responses indicated that participants engaged in ingroup favoritism, not to maximize differences between ingroup and outgroup but to improve ingroup members' gain. (3) Analysis of participants' open-ended self-reports revealed that they engaged in ingroup favoring reward allocation because of their personal bonds and likings for ingroup members as individuals rather than of their identification with the group per se. PMID- 12728699 TI - [Reading comprehension and working memory: structural equation modeling approach]. AB - We examined the relationship among intelligence, working memory, and reading comprehension using structural equation modeling (SEM). Ninety-six participants were instructed to perform two reading comprehension tests and six cognitive tasks: two verbal intelligence subtests, two spatial intelligence subtests, and two reading span tasks. Three latent variables that were called verbal ability, spatial ability, and working memory were derived from the six cognitive tasks. SEM demonstrated that the latent variables of working memory and verbal ability contribute to reading comprehension, suggesting that central executive functioning related to attention control was mediated among these cognitive abilities. PMID- 12728701 TI - [Do we talk to different people in different ways?: types and sources of social support and support-gaining strategies]. AB - The present paper examined whether people employ different support-gaining strategies toward various sources of support. In Study 1, 231 Taiwanese undergraduates were asked the frequency of each strategy they used for three support types: tangible, psychological, and informational, from four support sources: parents, professors, same-sex close friends, and same-sex acquaintances. In Study 2, 363 undergraduates were asked to think of an opposite-sex friend: an acquaintance, a close friend, or romantic partner, and write the frequency of each strategy they used. Results of ANOVA indicated that main effects of source and strategy and a three-way interaction of source by strategy by gender were significant. In Study 1, the students used various strategies most frequently toward parents and same-sex close friends, and least frequently to professors, and in Study 2, more frequently to close friends and romantic partners than to acquaintances. The strategy most often used was reasoning, followed by entreaty, roundabout request, exploitation, promise of reward, exhortation, and threat, in the descending order. No effect was found for the support type factor. PMID- 12728702 TI - [The lambda in the rorschach comprehensive system]. AB - The purpose of the present study was to investigate the relationships between Lambda (L) and other variables in the Rorschach comprehensive system. Individual Rorschach tests based on the comprehensive system was administrated to 102 female normal adults (over 20 years old). High L group and Low L group were defined on the basis of median value. The two groups were compared on major variables. The high L group had significantly higher values on variables such as DQo, D, Dd, S, Hd than the low L group. In contrast, the low L group had significantly higher values on variables such as DQ+, Blends, M, FM + m, Sum C', "active", H, COP than the high L group. Clinician who use the Rorschach test should consider these variables in relation to L, when interpreting the structural summary of the comprehensive system. PMID- 12728703 TI - [An item response model for the experimental design]. AB - In recent years, we see many examples of applying item response theory to psychological questionnaires. There are various conditions under which psychological tests are administered. Often, we are interested in the subjects' latent traits in the context of experimental design. In that case, we might want to know the effects of the experimental factors on the item parameters. In this study, we proposed an item response model in which items could be classified by some qualitative factors as in the situations where the items are administered under several different experimental conditions. The model's item parameters were linearly structured as in the general linear model. Then, we showed an example of analysis using an actual data set which was collected under four conditions, where the latent trait in question was "tension." PMID- 12728704 TI - [Relationship between encoding of the structural description system in implicit memory and binocular stereopsis]. AB - I investigated the relationship between encoding of the structural description system and binocular stereopsis. The structural description system is a component of the implicit memory system, and retains representations of possible figures. According to the previous studies, the priming effect based on it was produced only when the direction of the stimulus, which was the line drawing of the 3 dimensional object, was judged in the study phase. Because the judgment requires 3-dimensional processing, I proposed a hypothesis that the priming effect is also produced by binocular stereopsis in the study phase, and examined it by two priming experiments, in which different five undergraduates participated respectively. In the study phase, the primer was presented in binocular disparity (Experiment 1) or monocularly (Experiment 2), and the participants judged which lines were more, horizontal or vertical. In the test phase, the participants judged that the monocular stimulus was symmetrical or asymmetrical. The priming effect was produced only in Experiment 1, and was not in Experiment 2. The results supported the hypothesis, and suggest that encoding needs 3-dimensional processing in the structural description system. PMID- 12728705 TI - [Analyzing the factor structure and the sensory-relevance of impressions produced by words and drawings]. AB - The impressions of words and drawings that represented the same concepts (joy, anger, tranquility, etc.) were examined by the semantic differential technique. In the word (W) condition where the participants were presented with only the word stimuli, two factors (Evaluation and Activity) were extracted. By contrast, in the drawing (D) condition where the participants only saw the drawings, three factors (Evaluation, Activity, and Potency) were extracted and the potency factor was found to have large factor loadings for the adjective-pairs containing high relevance to tactile sensation. The same tendencies were found in the drawing + word (DW) condition where the drawings and the words were presented simultaneously. These results revealed that the factor structure in the W condition differed from those in the D and the DW conditions, and that the drawings aroused tactile sensations. Further analysis in the evaluation factor implied that the impressions produced in the DW condition had unique characters that could not be reduced to those in the W and the D conditions. PMID- 12728706 TI - Human osteoblast culture. PMID- 12728707 TI - Osteoblast isolation from murine calvariae and long bones. PMID- 12728708 TI - Mineralizing fibroblast-colony-forming assays. PMID- 12728709 TI - Osteocyte isolation and culture. PMID- 12728710 TI - Isolated osteoclast cultures. PMID- 12728711 TI - Primary isolation and culture of chicken osteoclasts. PMID- 12728712 TI - Isolation and purification of rabbit osteoclasts. PMID- 12728713 TI - Generating human osteoclasts from peripheral blood. PMID- 12728714 TI - Generating human osteoclasts in vitro from bone marrow and peripheral blood. PMID- 12728715 TI - Generating murine osteoclasts from bone marrow. PMID- 12728716 TI - Osteoclast formation in the mouse coculture assay. PMID- 12728717 TI - RANKL-mediated osteoclast formation from murine RAW 264.7 cells. PMID- 12728718 TI - Analysis of osteoclast function in mouse calvarial cultures. PMID- 12728719 TI - Assessing bone formation using mouse calvarial organ cultures. PMID- 12728720 TI - In situ hybridization and in situ reverse transcription polymerase chain reaction in human bone sections. PMID- 12728721 TI - Techniques for the study of apoptosis in bone. PMID- 12728722 TI - Protein localization in wax-embedded and frozen sections of bone using immunohistochemistry. PMID- 12728723 TI - Detection of noncollagenous bone proteins in methylmethacrylate-embedded human bone sections. PMID- 12728724 TI - Fluorescence imaging of bone-resorbing osteoclasts by confocal microscopy. PMID- 12728725 TI - Bone histomorphometry. PMID- 12728726 TI - Transmission electron microscopy of bone. PMID- 12728727 TI - Scanning electron microscopy of bone. PMID- 12728728 TI - Bone measurements by peripheral quantitative computed tomography in rodents. PMID- 12728729 TI - Studies of local bone remodeling. The calvarial injection assay. PMID- 12728730 TI - Inflammation-induced osteoporosis. The IMO model. PMID- 12728731 TI - Ovariectomy and estrogen replacement in rodents. PMID- 12728732 TI - Mechanical testing of bone ex vivo. PMID- 12728733 TI - Bone cell responses to fluid flow. PMID- 12728734 TI - Methods for analyzing bone cell responses to mechanical loading using in vitro monolayer and organ culture models. PMID- 12728735 TI - Extraction of nucleic acids from bone. PMID- 12728736 TI - Analysis of gene expression in bone by quantitative RT-PCR. PMID- 12728737 TI - Culture and psychiatry, or "the tale of the hole and the cheese". PMID- 12728739 TI - Why should researchers care about culture? AB - Analysis of the existing literature together with case experience reveals at least 4 implications of culture for the conduct of mental health research. Culture helps define the field of study, assists in identifying research gaps, shapes research paradigms, and supports the evolution of a cosmopolitan view of mental health. PMID- 12728738 TI - Cultural consultation: a model of mental health service for multicultural societies. AB - OBJECTIVES: This paper reports results from the evaluation of a cultural consultation service (CCS) for mental health practitioners and primary care clinicians. The service was designed to improve the delivery of mental health services in mainstream settings for a culturally diverse urban population including immigrants, refugees, and ethnocultural minority groups. Cultural consultations were based on an expanded version of the DSM-IV cultural formulation and made use of cultural consultants and culture brokers. METHODS: We documented the service development process through participant observation. We systematically evaluated the first 100 cases referred to the service to establish the reasons for consultation, the types of cultural formulations and recommendations, and the consultation outcome in terms of the referring clinician's satisfaction and recommendation concordance. RESULTS: Cases seen by the CCS clearly demonstrated the impact of cultural misunderstandings: incomplete assessments, incorrect diagnoses, inadequate or inappropriate treatment, and failed treatment alliances. Clinicians referring patients to the service reported high rates of satisfaction with the consultations, but many indicated a need for long-term follow-up. CONCLUSION: The cultural consultation model effectively supplements existing services to improve diagnostic assessment and treatment for a culturally diverse urban population. Clinicians need training in working with interpreters and culture brokers. PMID- 12728740 TI - Culturally competent psychotherapy. AB - To provide effective psychotherapy for culturally different patients, therapists need to attain cultural competence, which can be divided broadly into the 2 intersecting dimensions of generic and specific cultural competencies. Generic cultural competence includes the knowledge and skill set necessary to work effectively in any cross-cultural therapeutic encounter. For each phase of psychotherapy--preengagement, engagement, assessment and feedback, treatment, and termination--we discuss clinically relevant generic cultural issues under the following headings: therapist, patient, family or group, and technique. Specific cultural competence enables therapists to work effectively with a specific ethnocultural community and also affects each phase of psychotherapy. A comprehensive assessment and treatment approach is required to consider the specific effects of culture on the patient. Cultural analysis (CA) elaborates the DSM-IV cultural formulation, tailoring it for psychotherapy; it is a clinical tool developed to help therapists systematically review and generate hypotheses regarding cultural influences on the patient's psychological world. CA examines issues under 3 domains: self, relations, and treatment. We present a case to illustrate the influence of culture on patient presentation, diagnosis, CA, and psychotherapeutic treatment. Successful therapy requires therapists to employ culturally appropriate treatment goals, process, and content. The case also demonstrates various techniques with reference to culture, including countercultural, cultural reinforcing, or culturally congruent strategies and the use of contradictory cultural beliefs. In summary, developing both generic and specific cultural competencies will enhance clinician effectiveness in psychotherapy, as well as in other cross-cultural therapeutic encounters. PMID- 12728741 TI - Spirituality and religion in Canadian psychiatric residency training. AB - OBJECTIVE: Mental health professionals are increasingly aware of the need to incorporate a patient's religious and spiritual beliefs into mental health assessments and treatment plans. Recent changes in assessment and treatment guidelines in the US have resulted in corresponding curricular changes, with at least 16 US psychiatric residency programs now offering formal training in religious and spiritual issues. We present a survey of training currently available to Canadian residents in psychiatry and propose a lecture series to enhance existing training. METHODS: We surveyed all 16 psychiatry residency programs in Canada to determine the extent of currently available training in religion and spirituality as they pertain to psychiatry. RESULTS: We received responses from 14 programs. Of these, 4 had no formal training in this area. Another 4 had mandatory academic lectures dedicated to the interface of religion, spirituality, and psychiatry. Nine programs offered some degree of elective, case based supervision. CONCLUSION: Currently, most Canadian programs offer minimal instruction on issues pertaining to the interface of religion, spirituality, and psychiatry. A lecture series focusing on religious and spiritual issues is needed to address this apparent gap in curricula across the country. Therefore, we propose a 10-session lecture series and outline its content. Including this lecture series in core curricula will introduce residents in psychiatry to religious and spiritual issues as they pertain to clinical practice. PMID- 12728742 TI - Are mental health services for children distributed according to needs? AB - OBJECTIVES: The purpose of this study was twofold: 1) to determine whether publicly funded mental health services and resources available in 4 large regions in the province of Quebec were distributed according to the mental health needs of children aged 6 to 14 years and 2) to assess whether the variations in mental health services and resources across the 4 regions had changed over a 5-year period. METHODS: Indicators of need according to the child's parent (presence of mental disorder, measure of adaptation, and perception of need for help) from an epidemiologic survey of 2400 noninstitutionalized children were compared with both in-school and community professional resources and with physician and hospital services in 1992-1993. Resource and service data were also collected for 1997-1998. Resource and service data came from professional colleges and government administrative databases. RESULTS: No significant regional differences were found for need indicators, but there were large discrepancies in mental health resources and services in 1992-1993. Differences in professional resources were largest for special education teachers in the school system and for psychiatrists in the community. The regional differences in resources and services were as large in 1997-1998 as they were in 1992-1993. CONCLUSIONS: Despite universal health care in Quebec and a government mental health policy stressing equity of access, the available mental health resources for children aged 6 to 14 years are not distributed across regions according to needs. More evidence-based planning is required, specifically using epidemiologic survey data, to match resources to needs and to monitor changes over time. PMID- 12728743 TI - A random-assignment, double-blind, clinical trial of once- vs twice-daily administration of quetiapine fumarate in patients with schizophrenia or schizoaffective disorder: a pilot study. AB - OBJECTIVE: To evaluate the efficacy and safety of administering quetiapine once vs twice daily. METHOD: Utilizing a double-blind design, 21 hospitalized adult men or women with DSM-IV schizophrenia or schizoaffective disorder, who had received unchanged doses (for 2 weeks) of either 400 or 600 mg daily of quetiapine administered in 2 doses, were randomly assigned to once- or twice daily administration for 4 weeks and then crossed over to the opposite dosing regimen for an additional 4 weeks. Standard psychopathology and safety measures were used in the study. RESULTS: Nearly 70% (15/21) of the subjects met the a priori efficacy responder criteria with no statistical differences in response between those assigned to once- or twice-daily quetiapine administration. Statistical analyses confirmed that most subjects maintained efficacy during the switch to once- or twice-daily administration with quetiapine. A minority (15%) did experience worsening of symptoms or orthostatic hypotension during the crossover. Quetiapine was generally well tolerated at either twice- or once-daily administration. CONCLUSIONS: These pilot data suggest that it is clinically feasible to switch most quetiapine-treated subjects receiving a therapeutic twice daily dosing schedule to a once-daily regimen. A minority may experience worsening of symptoms or orthostatic hypotension during the switch. This strategy of administering quetiapine entirely at bedtime may promote improved adherence to treatment. PMID- 12728745 TI - Symptom outcome 1 year after admission to an early psychosis program. AB - OBJECTIVE: To determine the change in positive, negative, and depressive symptoms after 1 year in an early psychosis program. METHOD: One hundred and eighty subjects were included from the first 257 admissions for a first episode of psychosis to a comprehensive early psychosis program. Most had a diagnosis of schizophrenia or schizophreniform disorder. Subjects were assessed on admission to the program and at 3, 6, and 12 months after admission. All 180 subjects completed the 1-year assessment. Assessment measures included the Positive and Negative Syndrome Scale and the Calgary Depression Scale for Schizophrenia. RESULTS: There was a clinically and statistically significant improvement in positive symptoms by 3 months, depression increased at 3 months but significantly improved by 12 months, and negative symptoms changed little over the first year. CONCLUSIONS: The differential changes in symptoms in the first year after admission have implications for treatment. PMID- 12728744 TI - Essential fatty acids and the brain. AB - OBJECTIVE: To review the role of essential fatty acids in brain membrane function and in the genesis of psychiatric disease. METHOD: Medline databases were searched for published articles with links among the following key words: essential fatty acids, omega-3 fatty acids, docosahexanoic acid, eicosapentanoic acid, arachidonic acid, neurotransmission, phospholipase A2, depression, schizophrenia, mental performance, attention-deficit hyperactivity disorder, and Alzheimer's disease. Biochemistry textbooks were consulted on the role of fatty acids in membrane function, neurotransmission, and eicosanoid formation. The 3 dimensional structures of fatty acids were obtained from the Web site of the Biochemistry Department, University of Arizona (2001). RESULTS: The fatty acid composition of neuronal cell membrane phospholipids reflects their intake in the diet. The degree of a fatty acid's desaturation determines its 3-dimensional structure and, thus, membrane fluidity and function. The ratio between omega-3 and omega-6 polyunsaturated fatty acids (PUFAs), in particular, influences various aspects of serotoninergic and catecholaminergic neurotransmission, as shown by studies in animal models. Phospholipase A2 (PLA2) hydrolyzes fatty acids from membrane phospholipids: liberated omega-6 PUFAs are metabolized to prostaglandins with a higher inflammatory potential, compared with those generated from the omega-3 family. Thus the activity of PLA2 coupled with membrane fatty acid composition may play a central role in the development of neuronal dysfunction. Intervention trials in human subjects show that omega-3 fatty acids have possible positive effects in the treatment of various psychiatric disorders, but more data are needed to make conclusive directives in this regard. CONCLUSION: The ratio of membrane omega-3 to omega-6 PUFAs can be modulated by dietary intake. This ratio influences neurotransmission and prostaglandin formation, processes that are vital in the maintenance of normal brain function. PMID- 12728746 TI - [Memory is a faculty that does not remember]. PMID- 12728747 TI - Clinical and family history markers of bipolar II disorder. PMID- 12728748 TI - Effect of olanzapine on the liver transaminases. PMID- 12728749 TI - Protecting patients' health information: overview of the Health Insurance Portability and Accountability Act. PMID- 12728750 TI - The Hispanic/Latino Diabetes Educator Specialty. PMID- 12728751 TI - Nutrient intake, glycemic control, and body mass index in adolescents using continuous subcutaneous insulin infusion and those using traditional insulin therapy. PMID- 12728753 TI - Evaluation of diabetes management software. PMID- 12728752 TI - Math curriculum: an innovative approach to address weight issues in children. PMID- 12728754 TI - A randomized pilot study of improving foot care in home health patients with diabetes. AB - PURPOSE: The purpose of this study was to test the effectiveness of an educational intervention to improve patients' foot care knowledge, self-efficacy, and self-care practices. METHODS: A prospective, randomized, single center, 2 group design was used with a convenience sample of 40 home care patients from a Medicare-certified home health agency. Baseline measures of foot care knowledge, self-efficacy, and reported self-care practices were obtained at study entry and 6 weeks later to control for foot care interventions provided during routine home care services. After obtaining the 6-week baseline measures, patients who were randomized to the intervention group received individualized education about proper foot care. All patients were interviewed a third time 3 months after study entry to determine the effectiveness of the intervention. RESULTS: The educational intervention improved patients' knowledge, confidence, and reported foot care behaviors. CONCLUSIONS: A brief, individualized educational intervention about standard foot care topics improved patients' foot care knowledge and self-efficacy as well as reported self-care practices. Incorporating such interventions into routine home care services may enhance the quality of care and decrease the incidence of lower-extremity complications. PMID- 12728756 TI - Evaluating group visits in an uninsured or inadequately insured patient population with uncontrolled type 2 diabetes. AB - PURPOSE: This study was conducted to evaluate the feasibility and acceptability of a managed-care approach (group visits) on delivering care to uninsured or inadequately insured patients with type 2 diabetes. METHODS: One hundred twenty patients with uncontrolled type 2 diabetes were randomly assigned to receive care in group visits or usual care for 6 months. At baseline, 3 months, and 6 months, the feasibility and acceptability of this model of healthcare delivery were assessed through the patients' responses to the Primary Care Assessment Tool and the Trust in Physician Scale. Attendance records were kept for each group. RESULTS: Patients who received care in group visits showed an improved sense of trust in their physician compared with patients who continued to receive usual care. There was a tendency for patients in groups to report better coordination of their care, better community orientation, and more culturally competent care. Patient attendance at the groups also indicated good acceptance of this form of healthcare delivery. CONCLUSIONS: Group visits were feasible and acceptable to these uninsured and inadequately insured patients with uncontrolled type 2 diabetes and fostered an improved sense of trust in their physician. PMID- 12728755 TI - Adolescents' behavioral autonomy related to diabetes management and adolescent activities/rules. AB - PURPOSE: The purpose of this preliminary study was to describe and explore the behavioral autonomy (both independent functioning and decision making) of adolescents with type 1 diabetes. METHODS: A sample of 34 adolescents with type 1 diabetes completed checklists on independent functioning and decision making for daily and nondaily diabetes management as well as typical adolescent activities/rules. RESULTS: Independent functioning in daily diabetes management was greater for older adolescents. Independent functioning and decision making for daily diabetes management, nondaily diabetes management, and typical adolescent activities/rules were strongly correlated. Independent decision making, but not independent functioning for daily diabetes management, was significantly correlated to metabolic control. CONCLUSIONS: The strong relationship between independent decision making and functioning suggests that both aspects are important parts of behavioral autonomy to be assessed by healthcare professionals working with adolescents with type 1 diabetes. Healthcare professionals should encourage parental involvement that facilitates adolescents' independent decision making, which was related to better metabolic control in this study. PMID- 12728757 TI - Overcoming obstacles to behavior change in diabetes self-management. AB - PURPOSE: This study describes how 5 individuals with type 2 diabetes overcame obstacles that interfered with maintaining behavior changes in diet, exercise, and self-monitoring of blood glucose (SMBG). METHODS: In-depth interviews provided the framework for this descriptive study. During audiotaped, face-to face, follow-up structured and unstructured interviews, the participants shared their experiences about obstacles encountered and strategies used to overcome them. Each case was examined independently, and all cases were then compared with each other. RESULTS: Obstacles that interfered with maintaining a diet plan were hunger, planned and unplanned meal events, and the desire for new foods. Physical illness and unexpected life events were obstacles to maintaining an exercise and SMBG plan. CONCLUSIONS: Health professionals who assist patients who are beginning behavior change or having difficult maintaining changes need to have an understanding of the various obstacles and how patients can maintain behavior changes over time. PMID- 12728758 TI - College of Pastoral Supervision and Psychotherapy adds its sponsorship to the Journal of Pastoral Care & Counseling. PMID- 12728759 TI - Pastoral counseling and pastoral care: is there a difference? AB - The author argues that there is little difference between pastoral counseling and pastoral care. Utilizing an evidence-based and narrative approach, he examines the ideas of a variety of historical and contemporary writers to illustrate this thesis. Along with historical and contemporary writings on the topic, the author includes his own clinical experiences and associations to illustrate his conviction that pastoral counseling and pastoral care are more alike than different. PMID- 12728760 TI - First look: what brings clergy candidates into ministry and what happens when they don't get it. AB - This article reports on a retrospective examination of all candidates for ordained ministry in Alabama-West Florida Annual Conference of The United Methodist Church from September 1995 to 1998. The results showed, contrary to expectations, that clergy in this annual conference are significantly more likely than the general population to become physically ill when placed under stress. One of the major stressors identified in this study was the likelihood that the clergy persons will not receive the high levels of affirmation and reinforcement they look for. The author presents some conclusions and suggestions both for action to change the predictions and for further research. PMID- 12728761 TI - Clinical pastoral education: a physician's experience and reflection on the meaning of spiritual care in palliative care. AB - In this article a physician reflects on her experience as a chaplain intern and how this Clinical Pastoral Education experience led to a deeper understanding of spiritual care in the palliative setting. PMID- 12728762 TI - Blended families/Sarah, Hagar, and all that.... AB - A rabbi/chaplain listens to a 21st century woman tell about some complexities encountered in the blended family dynamics. He reminds her that such stressful phenomena are not as new as many believe, illustrating his point by interpreting the biblical account of Sarah and Hagar, noting particularly that some of the same decisions and actions taken in ancient times may still provide authentic insights to current family dynamics today. PMID- 12728763 TI - Homelessness. AB - The author, acknowledging the reality of homeless persons in most communities, explores the meanings and dynamics of homelessness, and the need to recognize the variety of participants needing to be recognized in appreciating the complexity of this segment of society. He raises the issue of how pastoral caregivers become involved in providing authentic care to this sub-culture and offers examples from his own experiences as a volunteer chaplain in the Salvation Army Corps. PMID- 12728764 TI - Dragon talk: providing pastoral care for Chinese immigrants. AB - This article describes how cultures and pastoral care education processes can be barriers between the patient, the pastoral caregiver, and the Clinical Pastoral Education (CPE) student. By providing sketches of interviews with Chinese patients, the author tries to explain why the attempt to unveil Chinese patients' feelings and needs through conversation can be a frustrating experience. Moreover, the author argues that the pedagogy of pastoral care education ought to be more culturally sensitive in regard to the diverse cultural backgrounds of both patients and CPE students. PMID- 12728765 TI - Rolling away the stone: post-abortion women in the Christian community. AB - This article seeks to bring to awareness and address the issues of a neglected group of people in need of pastoral care: women who have had an abortion. It offers some commentary regarding what many women experience following an abortion, focuses on the story of a specific woman, and goes on to offer a biblical perspective for a pastoral response to their needs. Responses to the article are offered by a healthcare chaplain and a seminary professor. PMID- 12728766 TI - Religion and health research: interpretation sends wrong message regarding need for hospital chaplains in health care institutions. PMID- 12728767 TI - Religion and health research: theological interpretations oversimplified. PMID- 12728768 TI - Religion and health research: critique of critique not well balanced. PMID- 12728769 TI - Developing an effective employee manual. PMID- 12728770 TI - Building a pyramid to practice success. AB - Although decisions about the goals of the practice originate with the dentist and drive every other system, a successful dental practice looks like a pyramid. The bottom level includes the effective management of time and resources through efficient scheduling and patient communication. The second level ensures the ability of the practice to reach its goals through results-oriented case acceptance scripting, patient finance options and advanced customer service. The third level focuses on creating long-term relationships with patients and turning the hygiene department into a "WOW" experience profit-center. Dentists must set practice management and production/profitability goals before all others. PMID- 12728771 TI - Making every day ideal. PMID- 12728772 TI - Protecting your office from employee theft. PMID- 12728773 TI - Strategies for winning the retirement game. The 412 (i) innovation. PMID- 12728774 TI - HIPAA privacy compliance deadline quickly approaching. PMID- 12728775 TI - Partner awareness regarding the adult sequelae of childhood sexual abuse for primary and secondary survivors. AB - This qualitative study investigates factors that may facilitate or impede awareness within couples regarding the sequelae of childhood sexual abuse for adult females and their partner. Six couples were interviewed about perceived effects of the abuse for self and partner and their perceptions regarding their awareness of these effects. Transcribed data were analyzed using grounded-theory methodology. Emergent themes regarding potential barriers to and facilitators of agreement are outlined in the context of the expressive and receptive abilities and motivations of each partner in communicating about the abuse. Preliminary implications for marriage and family therapy and further research are provided. PMID- 12728776 TI - Who's dragging their feet? Husbands and wives seeking marital therapy. AB - Despite its demonstrated efficacy, marital therapy's impact has been limited by couples' general reluctance to seek help until their problems become severe. To understand this delay, 147 married couples (294 individuals) in the process of seeking marital therapy were surveyed. Using multilevel confirmatory factor analysis, three relatively independent steps (problem recognition, treatment consideration, and treatment seeking) were identified. On average, wives were rated as completing all three steps before their husbands. Gender-role orientation, demographics, relationship satisfaction, and specific relationship problems (especially husbands' dissatisfaction with sex) were also predictive of the steps toward therapy. Implications for marital therapy are discussed. PMID- 12728777 TI - Making space for racial dialogue: our experience in a marriage and family therapy training program. AB - Marriage and family therapy (MFT) training programs need to create opportunities for all students to develop cultural competency by raising their racial awareness and sensitivity. Likewise, therapists of color need to be offered space in MFT programs to voice their experiences and venues for their voices to be heard. This article reports on the efforts within a master's level, accredited MFT training program to create space, through participatory action research, for the unique experiences of therapists of color and White therapists who are deeply committed to racial sensitivity. PMID- 12728778 TI - Parenting practices and the transmission of ethnic identity. AB - Three years after being interviewed, a nonrandom, purposeful subsample of 14 Jewish families from a larger sample of 48 families living in Central New York was reinterviewed. The primary aim of this follow-up study was to develop a descriptive understanding of parenting practices and the transmission of ethnic identity. Semistructured family interviews were conducted and coded using grounded-theory techniques, in particular the constant comparative method of analysis. Four main qualitative categories emerged from this study: Individual differences in teenagers, stages of ethnic identity development, parenting practices, and parental role models. Findings suggest that clear expectations, a type of authoritative parenting, could be associated with the positive transmission of Jewish ethnic identity. This type of parenting style was direct as parents expressed clear expectations for participation in Jewish activities both at home and in the community. PMID- 12728779 TI - Assessing interpersonal fusion: reliability and validity of a new DSI fusion with others subscale. AB - The Differentiation of Self Inventory (DSI) is a multidimensional measure of differentiation consisting of four subscales focusing on adults (ages 25+), and their significant relationships, including current relationships with family of origin. Although the DSI full scale and three of its subscales are theoretically and psychometrically sound, the Fusion with Others (FO) subscale is lacking. Therefore, responses of 225 adults were used to revise the FO subscale. Results yielded a 12-item, revised FO subscale with improved internal consistency reliability and construct validity. Greater fusion with others was associated with greater spousal fusion and dimensions of adult attachment insecurity. Implications for Bowen theory and suggestions for future research with the DSI-R are discussed. PMID- 12728780 TI - Family psychoeducation and schizophrenia: a review of the literature. AB - Family psychoeducation has emerged as a treatment of choice for schizophrenia, bipolar disorder, major depression, and other disorders. More than 30 randomized clinical trials have demonstrated reduced relapse rates, improved recovery of patients, and improved family well-being among participants. Interventions common to effective family psychoeducation programs have been developed, including empathic engagement, education, ongoing support, clinical resources during periods of crisis, social network enhancement, and problem-solving and communication skills. Application of family psychoeducation in routine settings where patients having these disorders are usually treated has been limited, reflecting attitudinal, knowledge, practical, and systemic implementation obstacles. Through consensus among patient and family advocacy organizations, clinician training, and ongoing technical consultation and supervision, this approach has been implemented in routine clinical settings. PMID- 12728781 TI - Affective disorders. AB - Unipolar depressive disorders are especially common among persons presenting with marital and family problems, rendering their impact on the treatment of marital and family problems of interest to every marriage and family therapist. The current paper describes empirically based decision making rules for determining who may benefit most from marital and family therapy interventions. Using well controlled outcome research, it is concluded that both marital therapy and parenting interventions can play an important role in the treatment of many but not all depressed persons. Predictors of treatment response are discussed and recent epidemiological research is used to examine the size of the population that may respond especially well to marital therapy. Future directions for research are suggested and public policy implications are discussed. PMID- 12728783 TI - Evaluating antipsychotic medications: predictors of clinical effectiveness. Report of an expert review panel on efficacy and effectiveness. AB - Over the course of the last decade, atypical antipsychotic medications (olanzapine, quetiapine, and risperidone) have become first-line choices for acute and maintenance treatment for schizophrenia. In numerous cases, these antipsychotics have replaced typical neuroleptics, which are associated with a high incidence of adverse effects, particularly parkinsonism and tardive dyskinesia. Each pharmaceutical company that produces an atypical antipsychotic has supported numerous studies in an attempt to demonstrate the superiority of its agent compared with its competitors. Many of these studies are, unfortunately, too brief in duration, given that schizophrenia and the other psychoses require long-term treatment. Nevertheless, those studies have yielded interesting and evidence-based data. The clinician cannot, however, rely entirely on data from studies. Clinical experience, which allows clinicians to follow patients for prolonged periods of time, is also important in choosing an antipsychotic medication. This article takes the refreshing approach of considering both of these aspects to guide more appropriate prescription of the atypical antipsychotics. PMID- 12728782 TI - The effectiveness of family interventions for physical disorders. AB - This article reviews the evidence for the effectiveness of family interventions in the prevention and treatment of physical disorders. Pathways by which families influence physical health and a typology of family interventions are described. Family intervention studies, particularly randomized clinical trials, are reviewed in four clinical areas: family caregiving of elders, childhood chronic illness, spouse involvement in chronic adult illnesses, and health promotion/disease prevention. Implications for family clinicians and recommendations for future research are presented. PMID- 12728784 TI - [Breakthrough in breast cancer chemoprevention]. AB - Breast cancer is the most frequent female malignant disease in developed countries. Various approaches are being developed for breast cancer prevention. Medical prevention called chemoprevention is reviewed. Prior to any intervention estimation of breast cancer risk is mandatory. For practical reasons distinction of two risk groups is useful. In the high risk group inherited gene mutation showing high penetrance may be suspected, while in the medium risk group hormonal factors play an important role. The antiestrogen tamoxifen has been extensively investigated in breast cancer and also tested for the prevention of breast cancer. The results of four randomized tamoxifen prevention studies have been published. In the largest, American trial the number of invasive or "in situ" breast cancers was halved by tamoxifen. Particularly estrogen receptor positive and relatively good prognosis breast cancers were reduced. Similar results were obtained in the "International Breast Intervention Study". Tamoxifen has been registered for breast cancer prevention for high risk individuals in the United States. The Italian and English ("Royal Marsden Hospital") studies did not prove significant preventative effect for tamoxifen that may be explained by the characteristics of the study protocols and study populations. Increased rates of endometrial cancer, thromboembolic events and cataract were observed under tamoxifen treatment, especially over the age of 50. Prevention has an increased importance in gene mutation carriers. Besides prophylactic mastectomy and close surveillance tamoxifen and bilateral oophorectomy or the use of gonadotropin releasing-hormone analogs seem efficient in this group. Various new chemoprevention strategies are under testing. Raloxifene and the aromatase inhibitors show advantage in menopausal women, the retinoid fenretinide and the gonadotropin releasing-hormone analogs seem promising for premenopausal individuals. The use of these agents are investigated in clinical trials. It is likely that not one single method will be applied for breast cancer prevention in the future. Preferably individual prevention strategies based on individual risk assessment will be developed. PMID- 12728785 TI - [Stress proteins in medicine]. AB - Stress proteins or in other words heat shock proteins form an ancient defense system of our cells. They are necessary to prevent the aggregation of damaged proteins and to help their refolding after stress. Stress protein-assisted remodeling of protein structure is an important step of many cellular processes, such as protein transport, signaling and protein degradation. Stress proteins have a key role in many diseases. Thus they 1. protect our cells against the deteriorating effects of ischemia/reperfusion in myocardiac infarcts or in stroke; 2. protect transplanted tissues and organs; 3. act against the multiple damage of chronic diseases such as diabetes, or neurodegenerative diseases (Alzheimer's and Parkinson's disease); 4. participate in the etiology of several autoimmune diseases; 5. their activation, and role in antigen presentation can be used as an anticancer-therapy; 6. stress proteins increase longevity, and lastly 7. stress proteins act as a buffer of phenotypically silent mutations and may contribute to the onset of "civilizational diseases" (cancer, atherosclerosis, diabetes, etc.). In this review the authors also summarize the existing stress protein-related pharmacological approaches to cure a large variety of diseases. PMID- 12728786 TI - [Role of the renin-angiotensin system in the pathogenesis, clinical picture and treatment of diabetic nephropathy]. AB - Prevalence of diabetic nephropathy is increasing. Understanding of pathogenesis and clinical picture helps to manage this disease. Recent data of the research of this disease support that the renin-angiotensin system plays a pivotal role in the pathogenesis. Hyperglycaemia activates the renin-angiotensin system and induces transforming growth factor-beta expression. These both lead to glomerulosclerosis and tubulointerstitial fibrosis. Diabetic nephropathy develops earlier and progress faster in patients with DD or ID genotypes of angiotensin-I converting-enzyme gene. Angiotensinogen and type 1 angiotensin-II-receptor gene mutations may be also predisposing factors for diabetic nephropathy. All these factors can be responsible for the hyperfiltration, albuminuria, salt sensitivity, and hypertension, which are characteristic features of diabetic nephropathy. According to these, one can suppose that inhibitors of the renin angiotensin system are effective in the prevention and treatment of this disease. Evidence of clinical studies suggests that angiotensin-I-converting-enzyme inhibitors in type 1 diabetes can prevent overt nephropathy, decrease proteinuria, inhibit the loss of the glomerular filtration and decelerate progression. Angiotensin-II-receptor blockers exert the same effect in type 2 diabetic patients, and presumably angiotensin-I-converting-enzyme inhibitors have similar activity in this group of patients. That is why, in the case of intolerance of one class of drugs, the other should be substituted. Combination therapy of angiotensin-I-converting-enzyme inhibitors with angiotensin-II receptor blockers can be the choice of treatment in the future. PMID- 12728788 TI - [Rapidly developing bone destruction in a patient with type 1 Gaucher disease]. AB - The authors report on a 25 year-old male Gaucher patient who developed rapid bone deterioration. The patient was splenectomised at age 3 and had no signs and symptoms of bone involvement for more than 20 years. In addition to the infarction in the right tibia, severe bone necrosis was detected in components of the left hip. This report suggests that bone deterioration may develop insidiously at any time in Gaucher patients, and that long-term and regular follow up is warranted. PMID- 12728789 TI - ["Index of non-quotation"]. PMID- 12728787 TI - [Experiences with endoscopic thoracic sympathectomy]. AB - PATIENTS AND METHOD: Thoracic sympathectomy using thoracoscopy was performed in 38 cases on 35 patients from January 01. 1996, till December 31. 2000. In 3 cases bilateral sympathectomy was carried out. The youngest patient was 18, the oldest was 76 years old, the average age was 42 years. The indications for surgery were Raynaud syndrome, causalgia, post-traumatic sympathetic dystrophy, thoracic outlet syndrome combined with vasospastic syndrome, Buerger syndrome, obliteration of digital arteries, embolism and hyperhidrosis. This method was chosen if conservative therapy was unsuccessful. Laparoscopic instruments are particularly suitable for minimal invasive interventions. RESULTS: Authors describe their operative technique whereby the postoperative pain and also the duration of hospitalisation can be reduced, and the cosmetic result can be improved. In two cases conversion was the only choice due to pleural adhesions. Pneumothorax occurred in two cases, haemothorax in one case and transient intercostal neuralgy was seen in 3 cases. In four cases sympathetic activity returned during the follow up. Ceasing the sympathetic innervation dilates the arterioles of the skin, and the temperature of the skin increases. During follow up the complaints of the patients improved significantly, the progression became slower and clear improvement was found with instrumental investigations. CONCLUSION: This method can be recommended to every institute where the conditions for traditional laparoscopic surgery are given and staff is experienced in thoracotomy. PMID- 12728790 TI - [Gastro-enteropathic neuroendocrine tumors in diverse hereditary multiple tumor syndromes of multiple endocrine neoplasms (MEN)]. PMID- 12728791 TI - [Papillary microcarcinoma of the thyroid. Clinical implications and therapeutic strategy]. AB - The Authors examine their experience about clinical implications and therapeutic strategies on Papillary MicroCarcinoma (PMC) of the thyroid gland. Clinical charts of 412 patients, who underwent thyroid surgery, were analyzed. The Authors stress "incidental diagnosis", benign associated lesions and frequency of population presentation; they conclude that the total thyroidectomy is the procedure of choice with oncology validity. PMID- 12728792 TI - [Current indications of bilateral adrenalectomy in Cushing's syndrome: discussion of personal experience]. AB - Since 1975 we have observed 68 patients with Cushing syndrome, 48 out of which with an ACTH-independent disease and 20 with an ACTH-dependent one, due to hypophisary adenoma. Out of the latter group of patients, which had a relapse of the disease after a previous trans-sfenoidal procedure, 16 underwent an open bilateral adrenalectomy and 4 a laparoscopic monolateral or bilateral adrenalectomy. Patients from the former group had an open or laparoscopic adrenalectomy, or a bilateral adrenalectomy in case of the rare bilateral diseases, as McCune-Albright syndrome. All patients had a complete healing, except for 2 patients affected by McCune-Albright syndrome who died for cardiopulmonary complications. We conclude that bilateral adrenalectomy plays a fundamental role in the treatment of Cushing disease, after the failure of the trans-sfenoidal procedures. This surgical therapy has been certainly improved by the introduction of the laparoscopic techniques, which allowed us to exert bilateral adrenalectomies in two times with a significant reduction of perioperative morbidity and mortality because of the less severe traumatism and the absence of addisonian complications. PMID- 12728793 TI - [Endoscopic approach to biliary stones: experience acquired in a general surgery unit. Comparison of 2 periods]. AB - The outcome of the laparoscopic technique, that in the first years needed to be applied in non complicated situations, imposed a more frequent use of ERCP preoperatively; this procedure was originally confined to a handful of European and American centers, but later spread to almost all large hospitals. Improvements in the techniques and materials have gone side by side with more specific indications and the assessment of complications. The purpose of the present study was to analyze the experience of a General Surgery Unit in terms of acquiring and developing skills in treating biliary stones by ERCP. PMID- 12728795 TI - [Duodenal villous adenoma: a case report]. AB - Small bowel tumours are rare neoplasms representing just 1% of all the gastrointestal tract lesions. Adenomas are the benign forms which, developing in the duodenum, tend to localize in periampullary region, with high incidence of malignant degeneration. The authors report a case of a fifty-five-years old woman with a periampullary villous adenoma. PMID- 12728794 TI - [Strangulated pelvic internal hernia: early postoperative complication of sacral colpopexy with prosthesis]. AB - The Authors report a case of internal strangulated hernia in a patient operated of colposacropexy for vaginal prolapse following total hysterectomy. The diagnosis was formularized by abdominal TC consenting the surgical treatment by bowel loop resection completed by pelvic floor reconstruction. PMID- 12728797 TI - [Familial adenomatous polyposis: review of the literature and report of 3 cases]. AB - The Authors describe three cases of Familial Adenomatous Polyposis, (FAP), in patients of the same family, mother and two daughters, with different stages of the disease. Familial adenomatous polyposis is a mendelian dominant inherited syndrome with an incidence of 1:11,000, caused by an alteration of APC gene, which causes multiple disorders of the development ecto-, endo- and mesoderma. The syndrome is characterized by the presence of adenomatous polyps in the gastroenteric tract, mostly in colon-rectum and duodenum with demonstrated adenoma-carcinoma sequence. In the family here reported a case of familial adenomatous polyposis at the adenomatous stage and two of cancer of colon-rectum are registered. In the first case surgery had a preventive aim, and ileo-rectal anastomosis was performed; in the other two cases the treatment was Miles operation with radical intention. PMID- 12728796 TI - [Small intestine tumors: our experience in emergencies]. AB - Small bowel tumours are relatively rare neoplasms; unusual occurrence associated with nonspecific symptoms, and low-sensitivity tests availability, are responsible for diagnostic delay. A retrospective study was performed on 42 cases with acute presentation, from 1972 to 2001; median age was 52 years (range 14-79) and there was a slight female prevalence (57.1% vs 42.9%). The most common acute presentation was occlusion (57.1%), followed by gastrointestinal (GI) bleeding (23.8%), perforation (14.3%) and occlusion/perforation (4.8%). Benign neoplasia were 38.1% (16 cases) and adenoma is was the most common type; malignant forms were 61.9 (26 cases) and adenocarcinoma and lymphomas were the most common histotype. Radical surgical procedures were possible only in 57% of malignant forms (24 patients); morbility was 4.8% (2 cases: 1 anastomotic dehiscence and 1 subphrenic abscess); mortality was 14.3%. From our retrospective study, we can state that survival for malignant lesions is strictly dependent of early TNM staging and possibility of radical surgical procedure. An extremely high index of suspicion in evaluating mild and often misleading symptoms, integrated with specific diagnostic studies, should be the proper approach. Prognosis for benign from is excellent in all cases. PMID- 12728798 TI - [Inguinal hernioplasty: Shouldice vs. tension-free techniques. A personal experience]. AB - The Authors report their experience with the use of Shouldice technique and tension-free procedures (Lichtenstein, Trabucco). They evaluate long time results with both procedures making a comparasion of Shouldice versus tension-free. The results evaluation underlines the best performance of tension-free procedures with elimination of recurrence. The Authors achieve that the tension-free techniques are now the gold standard for surgical treatment of groin hernia. PMID- 12728799 TI - [Abdominal Wells rectopexy with patch micromesh PTFE (Goretex)]. AB - The aim of this study is to discuss the surgical treatment of rectal prolapse. The surgical therapy offers many different options. Beginning from 1970 the anterior rectopexy has been considered a technique with good results, also in the elderly patient, when the general conditions allow it. Author's experience concerns the surgical treatment of 4 patients (2 males and 2 females) with complete symptomatic rectal prolapse, treated with anterior access laparotomy with the technique of Wells, leaving 1/3 of anterior rectal wall free from suture. Patients had a regular postoperative course; only in 1 case there was a delay of the canalization. At 1 year after the rectopexy the results are satisfactory, with absence of recurrence and troubles of the evacuation. The high percentage of successes of the anterior rectopexy is founded on: 1) the mobilization of the rectum, complete or only posterior; the rectal vessels must be respected, but there isn't consent on the opportunity of the section of the lateral ligaments. It may cause a denervation of the anorectum with an alteration of the fecal evacuation; and 2) the process of cicatritation by the employment of mesh and PTFE material that fix the rectum to the sacred maintains it as in the desired position. PMID- 12728800 TI - [Performance evaluation: dialogues and representations of a nursing group]. AB - This is a qualitative study, which aims at identifying the representations of performance evaluation through the point of view of those who evaluate and those who are evaluated. Nurses, technicians and nurse assistants of a school hospital participated in the interviews and focal group. Results showed that evaluation which focused on practices, generated suffering, and the ones, which focused on the being, constituted a source of satisfaction and development for those involved in the process. We concluded that, despite the broadest social imperatives, it is possible to redirect attitudes in the work routine. Considering this perspective, performance evaluation can be a very important tool for developing new practices. PMID- 12728801 TI - [Eye involvement in Hansen disease: evaluation in nursing consultations]. AB - This is a quantitative, exploratory-descriptive study. It analyzed patients who suffered from Hansen's disease based on social-demographic data, clinic form and time of disease evolution. It identified ocular alterations, relating clinic form to the time of evolution of this illness. The sample was composed of a group 60 patients: 39 were men and 21 were women, 35 were tan, 19 were white and 6 were black. The examination of these patients included eye-lid muscular power, external structures, ocular mobility, cornea sensitiveness, tear production, field and visual accuracy. The clinic form was dimorphous (42); virchovian (13); undefined (3) and tuberculoid (2). Disease evolution was from 1 to 4 years in 42 of the patients. Regarding eye alterations there was a reduction of tear production (32); of eye-lid muscular power (12) and of cornea sensitiveness (10); hyperemia (13); lagophthalmos (8) and madarosis (7), mainly concentrated in the dimorphous form (43) and virchovian (15). The multibacillaries present more intense ocular problems and it was not possible to establish correlation between them and time of evolution. PMID- 12728802 TI - [Pregnancy in adolescent women: the point of view of the relatives]. AB - This is descriptive study, which aims at understanding the family members' point of view, regarding the pregnancy of an adolescent. The research was carried out in Mossoro-RN, in Conjunto Liberdade II. Fourteen families participated in the study and the sample of family members was constituted of nineteen people: thirteen mothers, a grandmother, two mothers-in-law, a father, a stepfather and a father-in-law. Among the results of the study, it was verified that the preoccupation with physical and emotional immaturity of the pregnant adolescents, as well as economic matters dominated in the discourses of a considerable number of family members; that the situational intrafamily crisis seemed to be solved with the evolution of pregnancy and the conflicts could be resolved earlier when the adolescent was able to establish a consensual union with her sexual partner. It was also observed that, in spite of the crisis the family was going through, solidarity among the family members overcame the negative emotional reactions and the initial interpersonal conflicts. PMID- 12728803 TI - [Perceptions of adolescent women on maternity]. AB - The present study has as its objective to learn about the perceptions of adolescent mothers, regarding maternity in this stage of their lives, the health assistance given to them by the hospital team, and their expectations in relation to this assistance during the maternity period. This is an exploratory descriptive study with a qualitative approach, carried out with 11 puerperal adolescents during their stay an obstetric unit in a school hospital. Data were collected through semi-structured interviews and submitted to a theme analysis. The information gathered surfaced four main themes: 1) I am pregnant!, 2) Now, I have new responsibilities..., 3) Experiencing childbirth; To be mother is..., 4) We feel comfortable with the professionals here.... According to patient's reports it is feasible to have a humanistic assistance. In the specific case of the maternity analyzed the professionals were sensitive and compromised with the improvement of maternal health. PMID- 12728804 TI - [The nursing work process: the gap between planning and care delivery]. AB - This article aims at characterizing nurse's and nursing assistant's work process, as well as at analyzing contradictions present in nursing work. Studies on the nursing and health work process, and the communicative action theory were used as its theoretical framework. Based on qualitative research methodology, the empirical material was collected by means of semi-structured interviews with nurses, nursing assistants and physicians on the same work team in a school hospital. Analysis was conducted through the impregnation technique and the identification of thematic units, which were interpreted on basis of analytical categories from the theoretical framework. Results showed contradictions such as the separation between planning and care delivery and the absence of appropriation of nursing knowledge by the agents who are in charge of delivering care. PMID- 12728805 TI - [The Pro-ABEn project: contributions and reflexions for a marketing strategy for the association]. AB - In the present study, an internal analysis of the Brazilian Association of Nursing (ABEn) was carried out in order to identify the representations of the members of the national and regional management departments of ABEn, regarding this association. The objective was to gather subsidies for the planning and implementation of a project aiming at institutional empowerment. The scenarios for data collection were the regional and national management departments, as well as all the members available from the each regional section and two members from the national office. Data collection was done through a form, which was answered by 15 sections during the period between 1997 and 2002. The analysis of the empirical data material was done in two ways. The first analysis was based on the Discourse of the Collective Subject--DCS, and the second through Institutional Analysis. The representations of the subjects interviewed could be identified, enabling the planning of marketing strategies for ABEn. PMID- 12728806 TI - [Pregnant adolescents in a transition phase: a nursing focus on caring researching]. AB - The objective of this study is to understand the change women go through in their daily life, considering the transition during adolescence and maternity. It aims at understanding the existential meaning of the pregnant adolescent, facing transition, as a located phenomenon, in the simultaneity of life events. Data were collected through the discourse of eight pregnant adolescents who answered a phenomenological interview guided by the question: "What is it to be a pregnant adolescent undergoing transition, as a located phenomenon, in the simultaneity of life events?" The comprehensive analysis of the reports was based on the Heideggerian hermeneutics. It showed that pregnant adolescent experience this transitional moment with fear, fragility and ambiguous feelings of pleasure and suffering, according to the meanings attributed to their experiences. PMID- 12728807 TI - [Validation of the sterilization procedure of medical and hospital devices according to different packaging types]. AB - Safety in the processing of medical and hospital devices in healthcare organizations is an important measure of nosocomial infection control. This investigation aimed at establishing the period during which it is safe to use medical and hospital articles processed through sterilization by saturated steam under pressure using different types of packaging at a private hospital. The methodological procedure consisted of four phases: preparation of articles, evaluation of autoclave functioning, sterilization cycle and microbiological tests. Results showed bacterial growth on articles packed in surgical grade paper with and without film on the 21st day, in crepe paper on the 90th day and absence of growth when raw cotton packaging was used. In face of the results, a 21-day period was established for the use of articles after sterilization by saturated steam under pressure regardless of the type of packaging utilized, considering that storage conditions were also evaluated. PMID- 12728808 TI - [Experiences of women who undergo mastectomy: clarification and guidance]. AB - Based on a phenomenological approach, this study aims at understanding the meaning women with breast cancer attribute to the experience of excision of the breast. The comprehensive analysis of the report of seven women, revealed that there is a lack of assistance, especially due to denial of information, clarification and guidance regarding the problem. The study indicates the need of an assistance that takes into consideration the physical, psychological and social aspects of the patients undergoing this process, as well as their involvement with the multidisciplinary medical team. PMID- 12728809 TI - [Meta-cognitive strategies: a possibility in nursing education]. AB - This article is a theoretical reflection about the possibility of the application of metacognition strategies in nursing education. Metacognition is a reflective attitude of self-awareness and control of the mental processes. The metacognition area of cognitive psychology is analyzed, focusing on the problem solving process. The importance of metacognition in education is approached with emphasis on the teacher's role as a mediator in the student's learning process, in which "learning to learn" is emphasized. Some examples of its use in different situations of nursing education such as in text reading and nursing process, are reported. PMID- 12728810 TI - [Communicative action, education, and knowledge: an approximation to the Habermas thinking]. AB - This paper has the objective of presenting a synthesis of Habermas thought on communicative action, relating this with formal education, and more specifically with nursing education. Initially, different concepts of truth and knowledge, as well as the characteristics of modern age education, are pointed out. Secondly, the language concept, which serves as the base to Habermas communicative action theory, is presented. Finally, the study presents the contributions of this author to the materialization of a dialog-based form of teaching learning. PMID- 12728811 TI - [Technological education: why it is not relevant to the health field]. AB - This study discusses the National Curriculum Policies for professional education in technology, and its implementation in the health field, more especifically in the nursing field. The matter is approached from three different topics: 1) the contextualization of technological education, 2) the legal rationale, 3) the creation of human resources for SUS (Public Health system). As a conclusion, the study points out that the nursing professional would have his/her work even more fragmented than it already is. According to the study, this kind of education does not meet the needs of assistance in the present days, considering the complexity of problems related to it, the advancement of knowledge, and population growth. PMID- 12728812 TI - [Impact of a ministry edict: social and political aspects of obstetric nursing]. AB - This qualitative study presents the impact of the Ministerial Edict (number 2.815/98) relative to the inclusion of a set of procedures created by the obstetrician nurses in Public Health System (Sistema Unico de Saude--SUS). The objectives of this study are: present two excerpts which were published in a very popular newspaper and in a bulletin of the medical professional segment in 1998; analyze TV news which constitute different and tense discourses. The study identified two ideas, which were based on three pieces of news collected from the press. The first idea corresponds to the lack of visibility regarding the technical competence of obstetrician nurses in the social context. The second is related to the political acknowledgement of this service, which can be observed through the payment of obstetric nursing procedures according to a price list established by the Public Health System. Our final considerations are related to the social and political situation of obstetric nursing, in spite of the interventionist manipulation of society carried out by the media, which tries to create social-ideological discourses in order to obtain consensus and (re) produce truths. PMID- 12728814 TI - [Group intervention with a focus on emotional care: report of an experience]. AB - Attention to emotional health should be given at any circumstances of human beings development. It is especially necessary when an individual is ill and feels fragile, afraid, uncomfortable and anxious due to hospitalization. In this paper, we present an experience in which group work was used as a strategy of assistance to patients in the hospital, aiming at the implementation of an emotional assistance plan. The trajectory of the group, as well as the characteristics of the work done and its peculiarities are shown in this study. Results indicate that group work is a valuable tool for planning emotional care to these patients. However, it requires specific preparation and knowledge of group dynamics on the part of nursing professionals, so that the "healing" potential of group work can be better used. PMID- 12728813 TI - [Rescue of natural delivery in the technology era]. AB - This article discusses natural delivery, in spite of the technological development in the obstetric field. It proposes that natural delivery can generate a more humanized pregnancy and delivery and that it is an alternative proposal to the current model. Medical technology interference, in the delivery process, is associated to economic power, and presupposes that a safe delivery is the one done in a hospital provided with high technology. The current philosophy of medical delivery with severe asepsis and technical procedures, which are often unnecessary, interfere in this wonderful conquest of the evolution process that is the regular delivery. PMID- 12728815 TI - [Worldwide view of health and home care]. AB - Worldwide overview of health and home care. It describes the World Health Organization (WHO) in its historical background, pointing out the Brazilian participation in its foundation and some of the most relevant facts and events that preceded its creation. Also, it depicts the history of the Pan-American Health Organization (PAHO) founded in December 1902 and its celebration of the first centennial. A general overview regarding the world's health is given including globalization, as an irreversible fact, demographical and epidemiological changes, dependence and care delivery, home care services and policies on home care. PMID- 12728816 TI - [Group work: experiences of nursing students]. AB - This study aims at understanding the meaning attributed by undergraduate nursing students to the experience of group/team work. Based on the phenomenological framework. Individual interviews were conducted with nine students from the fourth year of the undergraduate program at the University of Sao Paulo at the Ribeirao Preto College of Nursing. Later, these were analyzed according to Martins and Bicudo (1989). The following categories emerged: group work focused on subjects; group work experienced in the classroom; group work: relationship among students and among students and teachers; group work in the training field; are students prepared for group work? By taking into account that professional practice in healthcare has shown the need to be interdisciplinary and that academic education presents gaps, concerning the practice of group work, re thinking about such education becomes relevant, since it will recover the understanding of complexity in human relations involving their political, institutional and interpersonal dimensions. PMID- 12728817 TI - Blurring the lines: expanding learning opportunities for children and youth. AB - This chapter explores why expanding commitments to learning is critical and presents a framework for broadening definitions of learning. PMID- 12728818 TI - Multiple pathways to adulthood: expanding the learning options for urban youth. AB - Some learning environments show particular promise of creating pathways to postsecondary education, careers, and engaged citizenship for urban young adults from ages fifteen to twenty-four. PMID- 12728819 TI - Building learning-centered communities through public engagement. AB - Local education funds can effectively engage their communities to align resources, policies, and public will to provide children and youth with supportive learning environments. PMID- 12728820 TI - High school after school: creating pathways to the future for adolescents. AB - This article proposes a strategy for the use of out-of-school time for adolescents, illustrates this strategy in action in Chicago, and discusses the challenges it faces. PMID- 12728822 TI - Blurring boundaries: the promise and challenge of a district-community action plan for systemic high school change in Worcester, Massachusetts. AB - The Worcester, Massachusetts, district-community plan for reinventing high schools shows how learning can be enhanced and transformed through intentional connections to community. PMID- 12728821 TI - Optimizing out-of-school time: the role of free-choice learning. AB - Free-choice learning, a new paradigm for the learning that youth and their families engage in outside school, can play an important role in the healthy development of youth, their families, and communities. PMID- 12728823 TI - Community as text: using the community as a resource for learning in community schools. AB - The community can serve as a resource to help students become more engaged in learning and strengthen connections between schools and community. PMID- 12728824 TI - Learning with excitement: bridging school and after-school worlds and project based learning. AB - A bridging typology captures a range of ways that after-school programs connect children's diverse worlds and support project-based learning. PMID- 12728825 TI - Mis-takes across paradigms. AB - In this column, the authors reflect on mis-takes across paradigms, misconceived or misunderstood meanings given and taken among scholars with differing worldviews. Mis-takes of paradigmatic perspectives, human science, distinctiveness, ways of knowing, and rationality are explored. It is proposed that mis-taken views of opposing perspectives are often deliberately advanced by scholars as ways to discredit the opposition, not to further dialogue. The authors call for an appreciation of difference in the context of the intellectual sparring and debating with regard to fundamental issues that characterize evolving disciplines. PMID- 12728826 TI - Mistakes in research: an appeal for tolerance. PMID- 12728827 TI - Uncovering meaning: how nursing knowledge changes policy in practice. AB - What ethical values, assumptions, and beliefs guide nursing practice today? What is the purpose and intent of policy in relationship to nursing practice? What underlying assumptions, values, and beliefs are missed in policy development that is not informed by nursing knowledge? These are some of the questions the authors attempt to answer in this paper. They examine some of the ethical dilemmas that arise when nurses are required to enforce policy that conflicts with nursing theory. A description of how nursing theory informs miss-takes (that is, missed assumptions, missed understandings, missed views and values) highlights how nursing knowledge changes policy in practice. PMID- 12728828 TI - Facing fear. PMID- 12728829 TI - Engaging the abyss: a mis-take of opportunity? AB - There are occasions when persons find themselves faced with a difficult choice to be with one or more persons as they live intense experiences that often involve pain and suffering. It is proposed in this column that the experience of choosing to be with others during these times calls forth a risking of self. That fleeting moment when people choose to engage with others or to turn away from that opportunity is explored through personal experience and as portrayed in poetry and film. The authors use the metaphor of an abyss to explore the lived experience of risking being with the truth of human connectedness. It is suggested that nurses know the abyss and that they have opportunities to be with persons as they live truth in human becoming. PMID- 12728831 TI - Theory and practice: a conversation with Marilyn E. Parker. Interview by Jacqueline Fawcett. PMID- 12728830 TI - Mis-takes: mistakes in the nurse-person process. AB - This column supports the global mantra to develop mechanisms that address the worldwide crisis in patient safety and demonstrates, using nursing as an example, that the mechanisms currently being developed are too limited. Patient safety initiatives must be expanded to acknowledge and make ways to minimize mis-takes, that is, misconceptions of meaning that emerge when healthcare professionals dismiss, misconstrue, guess, or undervalue patients' perspectives. Commitment to the human becoming school of thought as a guide for practice is suggested as one way to address mis-takes in the nurse-person process. PMID- 12728832 TI - A middle-range theory of caregiver stress. AB - The Roy adaptation model is a widely used nursing theory, but only a few studies have used it in the context of caregiving for the chronically ill. Many caregivers with chronically ill relatives suffer from stress and its consequences, and in an aging society, more and more people will be affected by stress. In this article, the author presents a middle-range theory of caregiver stress based on the Roy adaptation model as the first step in understanding the applicability of the Roy adaptation model in the context of stress in caregivers. PMID- 12728833 TI - Lingering presence: a study using the human becoming hermeneutic method. AB - The purpose of this human becoming hermeneutic research was to explore the meaning of lingering presence. The emergent meanings were: (a) A lingering presence surfaces in the cherished remembered which changes moment to moment as new experiences arise in the now and shed different light on the was and will be; (b) a lingering presence is lived in private ways, yet with others in a different alone-togetherness; and (c) a lingering presence is living with the familiar unfamiliar in the now moment, while moving beyond with different possibles. The findings are discussed in relation to how they contribute to nursing knowledge development. PMID- 12728834 TI - Experiencing health in the context of rheumatoid arthritis. AB - The purpose of this study was to explicate the health experience of persons with rheumatoid arthritis who manifest a strong sense of well-being. Five women and three men participated in unstructured interviews in order to answer the research question, What are the subplots that emerge in the descriptions of health given by persons with rheumatoid arthritis? A hermeneutic method was used to analyze the transcripts. Three subplots emerged: (a) The evolving--not knowing and then knowing something, but never really knowing, (b) the reframing, and (c) the relational fugue. The findings, including creation of healing spaces and an evolving relationship with assistive devices, provide evidence of turning points in the health process and are suggestive of a strong sense of well-being as a manifestation of a reorganized pattern. PMID- 12728836 TI - The meaning of the family: lived experiences of Turkish women immigrants in Germany. AB - The basic premise underpinning this research is that the family plays a key role in understanding and providing adequate nursing care for an ill member. To understand the family, the view on what members are as a family and what meanings are attributed to their various roles is of central importance for nurses. Since Turkish Muslim families who live as migrants in Germany differ considerably in their own understanding of themselves in their German environment, qualitative research, based on Spradley's developmental research sequence method, was conducted with nine Turkish women in order to learn about the meaning of family for them. PMID- 12728835 TI - Giving voice to persons living with dementia: the researcher's opportunities and challenges. AB - Few qualitative studies have examined the experiences of persons living with dementia and little has been written about the opportunities and challenges for the researchers. This article challenges the assumption of dwindling personhood among persons with dementia. Methodological challenges which may be encountered by qualitative researchers are outlined and specific strategies for addressing these challenges are described including: modifying consent/assent procedures, adjusting sample size, facilitating communication, and making the most of the data. PMID- 12728837 TI - Developing a nursing perspective on spirituality and healing. PMID- 12728838 TI - Lyrical language and nursing discourse. Can science be the cool of love? PMID- 12728839 TI - La Vie Boheme. PMID- 12728840 TI - Aripiprazole: quite possibly a new type of antipsychotic. PMID- 12728841 TI - Copper deficiency anemia and nephrosis in zinc-toxicity: a case report. AB - Zinc is a ubiquitous element that is essential for normal enzymatic function in multiple metabolic pathways. Chronic excessive zinc ingestion causes severe reversible anemia in humans. In animals, zinc toxicity leads to anemia as well as physiologic and morphologic damage to the pancreas, kidneys, and often, multisystem failure and death. In this case, a young female ingested approximately 2000 mg of zinc gluconate daily for 12 months. She subsequently developed anemia consistent with zinc-induced copper deficiency and severe nephrosis. After cessation of zinc ingestion, her anemia and nephrosis resolved. This case study underscores the importance of an accurate and thorough investigation of nutritional supplements during the history and physical examination. Given the promulgation of zinc for the treatment of skin disorders and the common cold, along with the commercialization of nutritional supplements, unimpeded by regulatory guidelines, it is imperative that primary care physicians be attuned to the potentially dangerous consequences of excessive zinc ingestion. PMID- 12728842 TI - Fellowship opportunity. CDC offers chance to work with national HIV leaders. PMID- 12728843 TI - National AIDS conference focuses on front lines of prevention, treatment. PMID- 12728844 TI - Congressional debate will determine funding for HIV efforts. PMID- 12728845 TI - Banking. Supreme Court upholds use of IOLTA accounts for needy. PMID- 12728847 TI - States unite to press for price concessions on AIDS drugs. PMID- 12728846 TI - HIV/AIDS epidemic poses serious threat to tuberculosis eradication. PMID- 12728848 TI - Drug cost. States concerned about Fuzeon. PMID- 12728850 TI - Pseudonym. Anonymity is at court's discretion in HIV disclosure case. PMID- 12728849 TI - Breach of duty. Blood test didn't create fiduciary duty to provide results. PMID- 12728851 TI - SSA disclosure of HIV status might have caused distress. PMID- 12728853 TI - Funding. HHS awards $600M to cities hardest hit by AIDS. PMID- 12728852 TI - Transmission. Study planned to track HIV spread among migrant workers. PMID- 12728854 TI - HHS awards portion of grants to minority HIV/AIDS services. PMID- 12728855 TI - Victim of attempted rape can seek HIV testing of attacker. PMID- 12728856 TI - Long path to approval. A look back at the road to Fuzeon. PMID- 12728857 TI - Fuzeon data review. PMID- 12728859 TI - $15 million for infrastructure but no AIDS drugs for Jamaica. PMID- 12728858 TI - FUZEON (enfuvirtide, T-20). Breaking barriers or breaking the bank? PMID- 12728860 TI - International treatment preparedness summit opens. PMID- 12728861 TI - It's time to face the Zerit problem. PMID- 12728862 TI - Drug resistance among HIV and its "friends". PMID- 12728863 TI - Skin infection spreads among gay men. PMID- 12728864 TI - Scientists link harmless virus to slowing of HIV's effects. PMID- 12728865 TI - Both sides of drug interruption debate supported. PMID- 12728866 TI - Histoplasmosis in a patient with AIDS. PMID- 12728867 TI - Truth, lies, and statistical tests. PMID- 12728868 TI - Current management challenges in HIV: antiretroviral resistance. AB - Emergence of drug-resistant viral variants is a major reason why HIV-infected patients experience viral rebound during antiretroviral therapy. Although combination antiretroviral therapy substantially inhibits viral replication, replication-competent mutant virus remains. In addition, it is now clear that virologic failure is not necessarily caused by failure of all drugs in a regimen. The use of resistance-testing data can assist in understanding the reasons for failure of antiretroviral therapy. However, there is a need for additional trials to better define the role resistance testing may play in developing management approaches to mitigate or minimize emergence of resistant HIV. PMID- 12728869 TI - Editorial comment: interpretation of resistance tests remains complex. PMID- 12728871 TI - Editorial comment: on the relationship between intracranial venous sinus thrombosis and HIV-associated nephropathy. PMID- 12728870 TI - Intracranial venous sinus thrombosis complicating AIDS-associated nephropathy. AB - An alert and oriented 27-year-old African American woman with AIDS presented with a 10-day history of fever, cough productive of yellow sputum, nausea, and vomiting and a 1-day history of excruciating headache and photophobia. Her condition rapidly deteriorated into a coma with decorticate and then decerebrate posture, and she died 3 weeks later. There was evidence of extensive intracranial venous sinus thrombosis (ICVST), renal vein thrombosis (RVT), and multiple cerebral hemorrhagic infarcts due to a hypercoagulable state complicating AIDS associated nephrotic syndrome. This is the first reported case of fatal ICVST and RVT with extensive cerebral hemorrhagic infarcts complicating nephrotic syndrome in a patient with AIDS. PMID- 12728872 TI - The promise and peril of HIV treatment. PMID- 12728873 TI - HIV testing and counseling in adult primary care settings. PMID- 12728874 TI - The answers continue. PMID- 12728875 TI - Prevention education: have we lost our focus? PMID- 12728876 TI - HAART-associated lymphadenopathy. PMID- 12728877 TI - Lipodystrophy: a new phenomenon? PMID- 12728878 TI - Hydroxyurea in the treatment of HIV infection. PMID- 12728880 TI - Treatment advances for CMV retinitis. AB - For many years the clinician's arsenal for managing cytomegalovirus (CMV) retinitis included only systematic therapies that required daily infusions via a central venous catheter. The introduction of cidofovir has permitted biweekly maintenance therapy by peripheral IV. Local therapies, such as the ganciclovir intraocular device and intravitreal injections of cidofovir, fomavirsen, or ganciclovir, have reduced toxicity while increasing absorption of drug in the eye. At the same time, due to highly active antiretroviral therapy (HAART), the incidence of CMV retinitis is dropping. Current investigation into the natural history of CMV retinitis in the era of HAART may ultimately result in new management strategies. PMID- 12728879 TI - Renal diseases in patients with HIV infection: a spectrum of outcomes in search of understanding. PMID- 12728881 TI - Oral mucosal herpes simplex ulceration in an HIV-seropositive man. AB - Herpes simplex virus (HSV) is a frequent cause of oral mucosal ulceration in HIV seropositive individuals. The case reported here illustrates the clinical presentation, diagnosis, and management of oral ulceration due to HSV. PMID- 12728882 TI - HIV-associated nephropathy: clinical characteristics and therapeutic options. AB - HIV-associated nephropathy is a unique form of renal disease specific to HIV infection. Proteinuria and rapidly worsening azotemia in the absence of edema and hypertension are characteristic. Renal biopsy reveals collapsing and/or sclerotic glomeruli, microcystic tubular dilatation, and cellular interstitial infiltrates. Direct cytopathic effects of HIV in the setting of a particular cytokine milieu appears to be the mechanism responsible for the renal injury. There is limited therapeutic experience, but trials with steroids and angiotensin-converting enzyme inhibitors are encouraging. Dialysis is the main form of renal replacement therapy. Better understanding of the disease should improve treatment and prognosis. PMID- 12728883 TI - The hypothalamus-pituitary-adrenal axis in HIV disease. AB - Abnormalities of the endocrine system, and of the hypothalamus-pituitary-adrenal (HPA) axis in particular, are associated with HIV infection. Opportunistic pathogens, neoplasms, and drugs used to treat infections may all contribute to the reported abnormalities, which range from subtle subclinical disturbances of HPA axis regulation to frank adrenal insufficiency. Patients with AIDS should be considered to be at high risk for primary or secondary adrenal insufficiency, and those with symptoms should be evaluated. Subclinical abnormalities may progress to clinically significant adrenal insufficiency as therapies improve and patients with AIDS live longer. PMID- 12728884 TI - Editorial comment: effects of HIV infection on the hypothalamic-pituitary-adrenal axis. PMID- 12728886 TI - Advances in the treatment of HIV dementia. AB - The onset of HIV dementia is uncommon until the middle-to-late symptomatic phases of HIV disease, when it may be found in up to 15% of patient populations. Signs and symptoms of dementia become progressively disabling. Highly active antiretroviral therapy may be effective in mitigating the degree of neurologic deterioration. Investigations of immune-based and neuroprotective agents as potential adjunctive therapies are under way or planned for the near future. Symptomatic treatment of psychiatric symptoms is an important adjunct to antiretroviral treatment of HIV dementia. PMID- 12728885 TI - Initiatives to promote adherence in HIV-infected patients. AB - Potent antiretroviral drug regimens have provided new opportunities to treat patients with HIV disease effectively. Yet, the true potential of the combination therapies can be realized only if people living with HIV adhere to the difficult treatment regimens. Even brief intervals of suboptimal dosing may allow activation of viral replication and development of drug-resistant strains. Maximizing adherence and achieving the full potential of the antiretroviral therapies in practice settings demand multidimensional initiatives that address complex behavioral and biomedical issues. PMID- 12728887 TI - Highlights of the 6th Conference on Retroviruses and Opportunistic Infections. PMID- 12728888 TI - Rx: energy infusion stat. PMID- 12728889 TI - Should ultrasensitive viral load assays be standard practice? PMID- 12728890 TI - Guidelines for managed care contracting. PMID- 12728892 TI - Common HIV drug-drug interactions. AB - Patients with HIV receive a multitude of medications daily, typically including a triple-drug antiretroviral regimen, prophylactic agents, and therapeutic drugs as needed. While the extensive drug regimens have resulted in improved survival and fewer opportunistic infections, they are also associated with the potential for undesirable and toxic drug interactions. The widespread use of protease inhibitors in particular has greatly increased the number and types of drug interactions that clinicians must monitor. PMID- 12728891 TI - Linear gingival erythema in an HIV-seropositive man. AB - Linear gingival erythema (LGE) is part of the spectrum of periodontal disease that exists in the HIV-infected population. Here, a typical case of LGE is presented and described. The microbiology and therapeutics of LGE are discussed, as well as this condition's relationship to more severe, rapidly destructive infectious diseases--such as necrotizing ulcerative periodontitis and necrotizing stomatitis. PMID- 12728893 TI - HIV prevention for teenage girls: relationships are the medium and the message. AB - HIV is a growing problem among young black and Hispanic women. Teenage girls' risk of HIV infection is primarily social in nature because sexual and/or needle sharing behaviors typically occur in the context of relationships. Without cure or vaccine, the practice of safe or protected personal behaviors is absolutely critical to prevent new HIV infections. To optimize HIV-prevention efforts, clinicians can form caring and developmentally appropriate relationships that address HIV education. Medical HIV prevention for teenage girls is fundamentally about healthy relationships. PMID- 12728894 TI - Prophylaxis and treatment of opportunistic infection in patients on HAART. AB - Opportunistic infections (OIs) are the hallmark of the immunodeficiency associated with HIV infection. Prophylaxis strategies have improved the morbidity and mortality associated with AIDS. The widespread use of highly active antiretroviral therapy (HAART) has brought about a change in the natural history and clinical manifestations of OIs; a decline in the incidence of most OIs; and an improvement in the clinical outcome of many OIs. The possibility of immune reconstitution raises questions about whether withdrawal or prophylaxis is safe and, in some cases, whether certain diseases may be curable without lifelong maintenance prophylaxis. Ongoing studies should provide information about the safety of removing primary and secondary prophylaxis in patients who respond to HAART. This review will update clinicians on the current state of the art of prophylaxis and treatment of OIs in patients taking HAART. PMID- 12728895 TI - Editorial comment: when to discontinue maintenance therapy for OIs in patients on HAART. PMID- 12728896 TI - HIV-associated Hodgkin's disease. PMID- 12728897 TI - Editorial comment: Hodgkin's disease in the setting of HIV. PMID- 12728898 TI - AIDS and emerging infectious diseases. PMID- 12728899 TI - Reports from the 6th Conference on Retroviruses and Opportunistic Infections. PMID- 12728900 TI - Which came first: social prejudice or fear of disease? PMID- 12728901 TI - Immune restoration disorders following HAART. AB - Highly active antiretroviral therapy (HAART) leads to a profound and sustained suppression of viral replication, along with a rise in CD4+ cells in most HIV infected patients. However, reports are accumulating of growing numbers of patients suffering from opportunistic infections despite recovery of CD4+ cells and plummeting viral loads as part of a new syndrome called immune restoration disease. We describe this syndrome in two patients and review the current literature. PMID- 12728902 TI - Kaposi's sarcoma in a nonadherent HAART patient. PMID- 12728903 TI - Women and HIV: when to test? AB - The devastating effects of HIV infection in women can be altered dramatically with new therapies. Certain clinical syndromes, including oral thrush, diffuse lymphadenopathy, wasting, unexplained fevers, or oral hairy leukoplakia, clearly indicate a need for HIV testing. This article reviews other clinical syndromes, both gender-specific and non-gender-specific, that may prompt the practitioner to consider HIV testing. PMID- 12728904 TI - Stevens-Johnson syndrome in an HIV-infected patient. AB - Stevens-Johnson syndrome (SJS) is a severe and sometimes lethal form of erythema multiforme most often associated with a drug hypersensitivity reaction. HIV infected patients suffer a higher incidence of SJS than the general population. This article details a case of nevirapine-induced SJS in an HIV-seropositive man and reviews the clinical features of erythema multiforme, SJS, and their association with drugs commonly used in the treatment of HIV infection. PMID- 12728906 TI - Editorial comment: HHV-6 infection in patients with HIV and other diseases: current status. PMID- 12728905 TI - HHV-6 infection in patients with HIV-1 infection and disease. AB - Human herpesvirus 6 (HHV-6) is among the most widespread of the human herpesviruses. In immunocompetent children, it causes exanthem subitum, febrile episodes without skin rash, and non-Epstein-Barr and non-cytomegalovirus infectious mononucleosis. HHV-6 has also been associated with clinical disease in bone marrow and solid organ transplant recipients. Its potential role in HIV-1 associated clinical syndromes is now being recognized and evaluated. In this review, we describe the virus, the pathogenesis of HHV-6-associated disease, and the diagnostic tests used to differentiate active from latent infection. We then discuss possible clinical manifestations of HHV-6 in HIV-1-infected patients, how to evaluate the need for treatment, and which pharmacologic agents are potentially useful. There is no consensus on these issues in the medical community, and HHV-6 is not now included among indicator infections for the diagnosis of AIDS. PMID- 12728907 TI - Anal dysplasia in men who have sex with men. AB - Risk factors for anal cancer include anal intercourse and infection with multiple strains of human papillomavirus, the causative agent of anal precancerous dysplasia. Several recent studies have shown that HIV-seropositive gay men are at greater risk for anal dysplastic lesions than seronegative gay men. Moreover, the risk for detection and progression of dysplastic lesions grows as the CD4+ cell count declines. A surgeon with a practice that includes gay men referred for anorectal disease presents data regarding the high prevalence of anal dysplasia in his patients. PMID- 12728908 TI - Tuberculosis is common in Ukrainian AIDS patients. PMID- 12728909 TI - Cidofovir. PMID- 12728910 TI - Understanding consciousness: implications and consequences. PMID- 12728911 TI - Environmental lead exposure and chronic renal disease. PMID- 12728912 TI - Environmental lead exposure and chronic renal disease. PMID- 12728913 TI - Imaging studies after a first febrile urinary tract infection in young children. PMID- 12728914 TI - Imaging studies after a first febrile urinary tract infection in young children. PMID- 12728915 TI - Hypertrophic cardiomyopathy and outflow tract obstruction. PMID- 12728916 TI - Hypertrophic cardiomyopathy and outflow tract obstruction. PMID- 12728917 TI - Preventing falls in elderly persons. PMID- 12728918 TI - Preventing falls in elderly persons. PMID- 12728919 TI - Regional anesthesia and analgesia for labor and delivery. PMID- 12728920 TI - Regional anesthesia and analgesia for labor and delivery. PMID- 12728921 TI - Indocyanine green angiographic features in endogenous Candida chorioretinis. AB - BACKGROUND: To describe indocyanine green (ICG) angiography (ICGA) findings and clinical features of endogenous mycotic endophthalmitis. PATIENTS AND METHODS: Two patients (a female 62 years, a male 31 years) were addressed to investigate a progressive unilateral visual loss. Slit-lamp examination disclosed a macular chorioretinitis. A clinical work-up revealed a mycotic infection (Candida albicans). Before treatment an ICGA was performed. RESULTS: ICGA early frames disclosed hypofluorescent lesions. Progressively, the lesions were surrounded by a slight hyperfluorescence, although the centre of the lesions was still hypofluorescent. CONCLUSIONS: The presence and persistence of a hypofluorescent lesion after introducing a specific treatment, led us to suspect a necrotic/ischaemic process affecting the choroidal vascular bed. ICGA provided additional information regarding the pathophysiological process and the patient's functional visual recovery. PMID- 12728922 TI - Design, construction, and in vitro analyses of multivalent antibodies. AB - Some Abs are more efficacious after being cross-linked to form dimers or multimers, presumably as a result of binding to and clustering more surface target to either amplify or diversify cellular signaling. To improve the therapeutic potency of these types of Abs, we designed and generated Abs that express tandem Fab repeats with the aim of mimicking cross-linked Abs. The versatile design of the system enables the creation of a series of multivalent human IgG Ab forms including tetravalent IgG1, tetravalent F(ab')2, and linear Fab multimers with either three or four consecutively linked Fabs. The multimerized Abs target the cell surface receptors HER2, death receptor 5, and CD20, and are more efficacious than their parent mAbs in triggering antitumor cellular responses, indicating they could be useful both as reagents for study as well as novel therapeutics. PMID- 12728923 TI - [Emergency medicine tomorrow]. AB - Emergency medical services (EMS) systems represent and important part within our health care system. However rapidly changing basic conditions define the need to reconsider current structures and objectives of the German EMS system and to implement appropriate modifications. The steadily decreasing number of emergency physicians advises to delegate more and more "technical" duties to paramedic personnel. In contrast the mobile intensive care unit will be reserved to cases in which complex treatment strategies or decisions have to be undertaken (i.e. thrombolysis). Moreover, the emergency physician's role will deeply expand beyond individual medical assistance. The most urgent duties Medical Advisers for Emergency Services will have to comply with are: 1. to develop plans of action and guidelines for paramedic personnel and 2. to secure a consistently high level of emergency medical care. As a consequence of ongoing structural reforms in the German Health Care System (ie. the implementation of diagnosis related groups, DRGs) emergency medicine will break down with traditional EMS structures. In contrast emergency medical departments of large hospitals will be reorganized to deliver both prehospital and in-hospital emergency medical care ("Centers of excellence"). Devoted to comply with clinical pathways this concept will improve medical efficacy as well as economic efficiency. The breakdown of traditional social structures as well as the increasing number of psychosocial emergencies constitute a major challenge to out EMS system. As it is not suitable to strictly differentiate medical from social emergencies we urgently need to establish integrated psychosocial networks. For a long period of time the role of the emergency physician has been restricted to the stabilisation of disturbed vital functions. To assure the competitiveness of the German EMS system it will be indispensible to expand our scope of practice. As anesthetists we should not hesitate to take over our new position as "managers in acute care medicine". PMID- 12728924 TI - The incidence, pathogenesis and natural history of steatorrhea after bone marrow transplantation. AB - A small number of case reports of steatorrhoea after allogenic BMT have been published, but the incidence and natural history of this complication have not been defined. We reviewed the incidence of steatorrhoea in 184 consecutive allograft recipients surviving at least 100 days. Steatorrhoea was documented in five patients, a median of 5.5 months (range 4-14) post-transplant. All patients had recent or concomitant acute gut or liver graft versus host disease (GVHD). The probability of developing steatorrhoea by 2 years post-transplant was 3.3% (95% confidence interval (CI) +/- 2.9% in the group overall, rising to 4.8% (CI +/- 4.2%) in patients with acute and/or extensive chronic GVHD. All patients responded clinically to pancreatic enzyme supplements. While these observations are consistent with previously reported autopsy data suggesting that GVHD of the exocrine pancreas is likely to be predominant underlying pathological process, in two patients concomitant small bowel or hepatic dysfunction may have contributed to the severity of steatorrhoea. Enzyme supplements were subsequently ceased in three patients without return of steatorrhoea, suggesting that the process is usually reversible. Our data demonstrate that steatorrhoea is not infrequent complication in the 2 years postallograft, particularly in patients with GVHD. PMID- 12728926 TI - Record reviews, clinician education form best defense. PMID- 12728925 TI - Pravastatin benefits elderly patients: results of PROSPER study. PMID- 12728927 TI - Introduction: Researching the impact of information technologies on radiology technologists: "taking it to the streets". PMID- 12728928 TI - Champions for consistency. PMID- 12728929 TI - Phonomicrosurgical techniques for treatment of RRP in children. PMID- 12728930 TI - Posterior surgical approaches to the elbow: a comparative anatomic study. PMID- 12728931 TI - Partial medial epicondylectomy for cubital tunnel syndrome: outcome and complications. PMID- 12728932 TI - [Dental care for patients with antineoplastic radiotherapy of the head and neck]. PMID- 12728933 TI - Fibrous connection to bone after immediate repair of the canine infraspinatus: the most effective body surface for tendon attachment. PMID- 12728934 TI - Bibliography:. Current world literature. Hematopoiesis. PMID- 12728935 TI - JCAHO forms Nursing Advisory Council. PMID- 12728936 TI - Current awareness on yeast. PMID- 12728937 TI - Introduction of erm(C) into a linezolid- and methicillin-resistant Staphylococcus aureus does not restore linezolid susceptibility. PMID- 12728938 TI - Patient notes: Hormone replacement therapy. PMID- 12728939 TI - Vista Award recipients. The best in team-based projects. PMID- 12728940 TI - [The irrigator Eguisier]. AB - Destinated, first, to cure gynecologic diseases, the irrigator Eguisier is the result of the work of two men: Maurice Eguisier (gynecologist) and Francois Libault (truss manufacturer). The patents and the trademark allowed the author to recall the commercial history of this machine which existed during the last fifteen years of the XIXe century. PMID- 12728941 TI - Orthopaedics in the developing world: present and future concerns. PMID- 12728942 TI - Toward a more natural medicine. AB - Modern science and culture attempt to dominate and control nature. Unfortunately contemporary medicine has followed this lead. Traditionally religion has recognized the order and goodness in the natural order and has encouraged followers to comply with the rules of nature. Medicine, from Hippocratic times, has viewed the physician as an assistant to nature. Contemporary medicine, while employing modern science, would be well advised to also recognize the healing effects of religion's admonition to acknowledge and respect the wisdom of nature. PMID- 12728943 TI - The arrogant physician--a Judaic perspective. AB - Arrogance among physicians is all too common. This is in sharp contrast with medicine's sacred religious origins. The author presents a concise historical review of the Judaic perspective on the role of the physician, his position in the community, and the attitudes/values that should characterize his healing pursuit. PMID- 12728944 TI - Bioterrorism, embryonic stem cells, and Frankenstein. AB - The stem cell controversy raises a fundamental question for humankind. Does science have a right to pursue knowledge whatever the cost? Our Enlightenment culture says yes. However, human history and literature are sending warning signals. Ethical issues impact the "knowledge for its own sake" imperative, and must be addressed. PMID- 12728945 TI - Amedeo Modigliani and Paul Guillaume: the "artiste maudit" and the "novo pilota". PMID- 12728946 TI - The Department of Neurosurgery at Seoul National University: past, present, and future. PMID- 12728947 TI - The sizer-dissector for aneurysm clip selection: technical note. PMID- 12728948 TI - The early history of neurosurgery in Brazil. PMID- 12728949 TI - Necrosis and glioblastoma: a friend or a foe? A review and a hypothesis. PMID- 12728950 TI - Brain death documentation: analysis and issues. PMID- 12728951 TI - Transsphenoidal treatment of empty sella by means of a silastic coil: technical note. PMID- 12728953 TI - Characteristics of human body dynamic responses to landing impact in supine position. AB - OBJECTIVE: To study the characteristics of human body dynamic responses to landing impact. METHOD: Five healthy young men were exposed to 4-10 G for 50-80 ms half-sine acceleration pulse at 20 degrees supine angle and 10 G for 50 ms impact at 30 degrees to 60 degrees supine angles. The acceleration curves on the dropping platform of the impact tower, the seat, the human head, shoulder, chest and ilium, as well as ECG of the subjects were recorded. RESULT: The peaks of acceleration at the subject head, shoulder, chest, and ilium along chest-back direction rose with the increase of the impact level. The overshoots of acceleration at the subject head along head-ilium direction and chest-back direction were 93% and 102% respectively for 10 G at 20 degrees supine angle. With increase of supine angle, acceleration response at the subject along chest back direction decreased. Acceleration response at the subject head along head ilium direction was minimum at 40 degrees supine angle. CONCLUSION: Transfer function of the human body to landing impact demonstrated that human body had a series of resonance frequencies which were 25 Hz, 34 Hz, 39 Hz, 43 Hz, 50 Hz, 64 Hz, and 70 Hz respectively. PMID- 12728952 TI - +Gz protection of a new bladder anti-G system. AB - OBJECTIVE: A new bladder anti-G system was developed to get better +Gz protection. The purpose of this study was to assess the +Gz protection afforded by it on the centrifuge. METHOD: Six male subjects using the new bladder anti-G system were exposed to 10-15 s ladder-shaped +Gz profiles without anti-G straining maneuver (AGSM), then exposed to + 5-9 Gz simulated aerial combat maneuver (SACM) with leg straining. RESULT: The +Gz protection and the +Gz-time tolerance were obtained. The anti-G effectiveness of the advanced bladder system was 5.33 +/- 0.26 G. All subjects passed the +5-9 Gz SACM, but the effort and fatigue were lower. CONCLUSION: The +Gz protection afforded by this system is high. The suggested scheme of the new bladder anti-G system is feasible. PMID- 12728954 TI - [Development and application of self-powered short arm human centrifuge]. AB - OBJECTIVE: To present the development and application of a self-powered short arm human centrifuge. METHOD: Self-powered short arm human centrifuge consisted of human power, adjustable resistance trig, short arm and supporting construction parts. It was driven by human power. The short arm turned around the supporting part so artificial gravity is produced. The physical load can be adjusted by the adjustable resistance part. G-level depends on the rotative velocity of the central shaft. Nine healthy male volunteers received self-powered short arm human centrifuge training for 5 min per day with rotative velocity of 30-34 r min-1 for 7 d. Cardiac pumping function was measured before and after training. RESULT: Heart rate (HR) decreased significantly while left ventricular ejection time (LVET) increased significantly after 3 d training, and HR and LVET changed further after 7 d training. Stroke volume increased significantly only after 7 d training. CONCLUSION: Self-powered short arm human centrifuge combines artificial gravity and exercise and included other advantages of safety and economy of construction. Cardiac pumping function could be improved by 7 d training. It may be used for anti-G physiological training or as a countermeasure to counteract the effect of microgravity. PMID- 12728955 TI - [Differential visual attention scales directed by location versus semantic cue]. AB - OBJECTIVE: To compare the cortical mechanisms associated with visual spatial attention directed by location cues and Chinese character cues. METHOD: Eleven healthy adults (mean age=19) with normal vision served as the subjects. In experiment I, the cue was a circle with black solid line. There were three cue sizes: small, medium and large. In experiment II, three black concentric circles were presented as background, their diameters were the same as the three cues used in experiment I. The cue was one of three Chinese characters small, medium or large respectively. The task of the subjects was to discriminate the target's orientation. RESULT: RT in experiment II was longer with short ISI. The anterosuperior N1 amplitude elicited by the cue in experiment II was larger than that in experiment I. The inferoposterior P1 latency in experiment II was longer than that in experiment I. Comparing the target evoked ERP, the inferoposterior P1 and N1, latency of experiment II was longer than that in experiment I, the inferoposterior P2 amplitude in experiment II was larger than that in experiment I. CONCLUSION: The processing of Chinese cue takes place at higher level functional brain regions compared to processing of location cue. The later requires more resource in the earlier stage of cue process and less in the subsequent stage of the task. PMID- 12728956 TI - [Effects of +Gz exposure time on memory and behavior in rats]. AB - OBJECTIVE: To explore the effect of +Gz exposure time on memory and behavior in rats. METHOD: Twenty-four male SD rats were randomly divided into control group (A), +10 Gz/3 min group (B) and +10 Gz/5 min group (C). Rats in group A and B were exposed to +10 Gz for 3 min or 5 min respectively. Changes of memory and behavior in rats were observed after +Gz exposure. RESULT: As compared with control, percentage of right reflex decreased significantly and reaction time lengthened significantly immediately, 1 d, 2 d, 4 d and 6 d after +10 Gz/5 min exposure (P<0.01); reaction time lengthened significantly immediately and 2 d after +10 Gz/3 min exposure (P< 0.01). As compared with +10 Gz/3 min, percentage of right reflex decreased significantly and reaction time lengthened significantly at 1 d, 2 d, 4 d and 6 d after +10 Gz/5 min exposure (P<0.01). As compared with control and +10 Gz/3 min, time stayed in center grille lengthened significantly immediately after +10 Gz/5 min exposure (P<0.01); balancing function decreased significantly immediately and 2 d after +10 Gz/5 min exposure (P< 0.01). CONCLUSION: It is suggested that +10 Gz/ 3 min exposure may induce a provisional disturbance in memory function and a change in behavior in rats, and +10 Gz/5 min exposure may induce a sustained disturbance in memory function and an obvious change in behavior in rats. PMID- 12728957 TI - [Cardiovascular effect of positive pressure breathing using G-suits with different bladder coverage]. AB - OBJECTIVE: To compare cardiovascular function of subjects exposed to positive pressure breathing (PPB) while wearing G-suits with different bladder coverage. METHOD: 6 male healthy subjects (18-20 years) wearing three kinds of G-suit with bladder coverage of 45%, 65% and 90% respectively, were exposed to PPB with counterpressures of 30, 50 or 70 mmHg for up to 3 min continuously. G-suit bladder pressure value was three times as high as that of PPB counterpressure. Cardiovascular function was observed every minute. The interval between PPBs was 10 min. The equipment consists of G-suit, jerkin, helmet and mask. RESULT: All subjects completed the experiment successfully. Mean arterial pressure (MAP) elevated significantly, while stroke volume had no striking change, as compared with control group when wearing G-suit with 65% or 90% bladder coverage G-suit. During PPB for 3 min, cardiovascular function didn't change significantly. The mean value of MAP was proportional to the level of PPB. CONCLUSION: The greater G suit bladder coverage, the less cardiovascular function effect when PPB was conducted simultaneously. PMID- 12728958 TI - [Effects of nutritional intervention on brain functions and its related biochemical indexes in mice exposed to high +Gy]. AB - OBJECTIVE: To study the effects of nutritional intervention on changes of monoamines in rats brain, certain amino acids in rats serum and maze behaviors caused by high +Gy exposure. METHOD: The mice were arranged into control group (Group A), +Gy group (Group B) and +Gy nutritional intervention group (group C). Group A was not exposed to +Gy. Both Group B and group C were exposed to +10 Gy for 8 min. Three hours before +Gy exposure distilled water was given to mice in group B by gavage. The day before exposure pyridoxal fortified water was given and 3 h before exposure mixed amino acids solution were given by gavage to group C. Maze test scores were recorded for all groups. After the maze test was completed, blood was collected through the eyes for serum amino acids and brain tissue was collected by decollation for monoamines and gamma-glutamyl transferase (GGT) activity determinations. RESULT: After +Gy exposure maze test scores and brain NE concentrations decreased and abnormal behaviors were found. While other monoamine transmitters increased significantly. In group C maze behaviors improved and biochemical changes induced by +Gy exposure alleviated. CONCLUSION: High +Gy exposure can induce changes of neural transmitters coming from nutritional metabolites in central nervous system. As a result, brain functions are affected. Nutritional intervention can alleviate this negative effect to some extent. PMID- 12728959 TI - [Study on mitochondrial membrane mechanism of rat heart injury induced by constrainting stress]. AB - OBJECTIVE: To investigate the effect of constrainting stress on biological characters and function of mitochondrial membrane in rat heart and to explore the possible mitochondrial membrane mechanism underlying stress-induced heart injury. METHOD: Stress animal model was established. After constrained for different times, all rats were killed and several indexes were examined. RESULT: Constrainting stress can induce mitochondrial permeability transition, decrease of mitochondrial membrane fluidity, increase of the production of membrane lipid peroxidation and injury of mitochondrial respiratory function which is in time dependent manner. CONCLUSION: Mitochondrial membrane impairment and its effects are the important mechanism of stress-induced heart injury. PMID- 12728960 TI - [Effects of microwave irradiation on ATPase activity and voltage dependent ion channel of rat hippocampus cell membrane]. AB - OBJECTIVE: To study the effect of microwave irradiation on hippocampus cell. METHOD: Changes of ATPase activity and voltage dependent ion channel of hippocampus cell membrane were observed in mice exposed to 2 450 MHz microwave irradiation of 10 mW/cm2 from a physical therapy machine. Histochemical method and patch clamp method were used to determine the activity of Na+, K(+)-ATPase, Ca2+, Mg(2+)-ATPase and voltage dependent Na+, K+, Ca2+ channels respectively. RESULT: 1) Na+, K(+)-ATPase activity of microwave irradiated mice showed no significant change as compared with the control, but the activity of Ca2+, Mg(2+) ATPase decreased significantly (P< 0.05); 2) In microwave irradiated mice, Na+, K+, Ca2+, current inducement rate in hippocampus neuron decreased significantly, the membrane voltage of Na+ current peak shifted to depolarization, and the attenuation rate of Na+ current and current A inducement rate decreased significantly as compared with control mice. CONCLUSION: Irradiation of 2 450 MHz microwave at a doze of 10 mW/cm2 was not fatal to mice hippocampus cell. But Ca2+, Mg(2+)-ATPase activity of hippocampal cell membrane and voltage dependent Na+, K+, Ca2+ ion channel of hippocampal nervous were affected which would affect study and memory. PMID- 12728961 TI - [Blood oxygen and lactate concentrations in skeletal muscles during exercise]. AB - OBJECTIVE: To study the relationship between blood lactate and blood oxygenation in human skeletal muscle. METHOD: Using a portable tissue oximeter based on Near Infrared Spectroscopy (NIRS), concentration of HbO2, blood lactate and blood volume were measured non-invasively and continuously when the subjects were doing incremental exercise on a bicycle ergometer. RESULT: As the intensity of exercise was increased, blood lactate concentration, blood volume in tissue increased, while concentration of HbO2 decreased. CONCLUSION: It is possible to assess the fatigue state with tissue oximeter by monitoring intensity of exercise non invasively and dynamically. By studying their correlations, a novel approach for measuring blood lactate non-invasively and assessing sports ability could be provided. PMID- 12728962 TI - [Analysis of heat transfer in skin tissues subjected to hot water]. AB - OBJECTIVE: To analyze heat transfer process in skin tissues subjected to hot water and thus to predict the extent of scald. METHOD: Basing on Pennes' Bio-heat Equation, a model for characterizing the damage process in tissues was presented and used for its analysis. RESULT: Temperature distribution in the tissues during the process and the time of beginning of scald were numerically obtained. It was found that the effects of different initial temperature distribution, convection coefficients between the hot water and the tissue surfaces, and the temperature of the hot water are significant. CONCLUSION: The transient heat transfer behavior in multi-layer tissues subjected to hot water can be predicted by the present model. It is useful in the clinical diagnosis and therapy for the scald caused by hot water. PMID- 12728963 TI - [The linear dimensions of human body measurements of Chinese male pilots in standing posture]. AB - OBJECTIVE: To provide the latest anthropometric data of Chinese male pilots on a large scale. METHOD: 94 linear dimensions of human body measurements were defined, of which there are 42 fundamental items and 52 recommended items. The computer databanks were programmed, in which the subprograms were preset for data checking such as extreme value examination, logical judgement for data relationship, and measuring-remeasuring difference test. All workers were well trained before pilot measurements. 1739 male pilots from China Air Force was measured for the 42 fundamental items, and of which 904 pilots were measured for the 52 recommended items. RESULT: Mean, standard deviation, the maximum value, the minimal value, and the 5th, 50th, 95th percentile data of all the 94 items were given. The quality of the data was stable and reliable. CONCLUSION: All data of the 94 linear dimensions of human body measurements were valid and reliable with high precision. PMID- 12728964 TI - [A study on visual search and selection of pull-down menu]. AB - OBJECTIVE: To explore the characteristics of user's visual search and selection through interaction with pull-down menus. METHOD: The visual search time and selection time of interaction were recorded by eye tracking system, when 14 participants were performing the task of pull-down menu selection, with different menu length, type of menu item and position of target item. RESULT: It was found that selection time was influenced jointly by menu length, type of menu item and target location, while visual search time was only influenced by target location. CONCLUSION: The results of this experiment support the hypothesis that the pull down menu was characteristic of serial, self-terminated and top-down. PMID- 12728965 TI - [Effect of extremely low frequency magnetic field on the focal brain injury in rats]. AB - OBJECTIVE: To examine the effect of extremely low frequency magnetic field (ELMF) on the brain trauma in rats. METHOD: Using the focal left-brain cortex contusion model, rats were divided into two groups, ELMF group (3 d and 8 d) and control group (3 d and 8 d), ELMF stimulations were given after the brain trauma provided by a coil (phi 10 cm) which was driven by 15 Hz sinusoidal signals so that ELMF strength of 18 mT at the midpoint was obtained. The morphologic changes were observed in these groups. RESULT: As compared with the control group, the extent of the inflammatory reaction and the neuronal damage was apparently lighter in magnetic field group, especially in 8 d group. At the far away region from the injured brain area, the neuronal shape was changed, and the amount of this special neurocyte was larger in magnetic field group than that in control group, especially in 8 d group. CONCLUSION: ELMF stimulation in the present study may alleviate the brain injury reaction. Attention should be paid to the role of the special neurocyte during the brain injury. PMID- 12728966 TI - [Encapsulated ANP cDNA transfection cells attenuate hypertension in hypertensive rats]. AB - OBJECTIVE: We Investigated a gene therapy delivery system based on microcapsules enclosing recombinant Chinese hamster ovary (CHO) cells engineered to secrete a therapeutic peptide-atrial natriuretic peptide (ANP). METHOD: Human atrial natriuretic peptide gene transfecting Chinese hamster ovary (CHO) cells were encapsulated in non-antigenic biocompatible polycaprolactone (PCL) capsules prior to their implantation into rats, then, the PCL-tubes were implanted into hypertensive DSS rats intraperitoneally. RESULT: The PCL-tubes 2 d post implantation caused a significant delay of blood pressure increase. The effect lasted for more than 5 months. The PCL-tubes also caused significant increases in renal blood flow, glomerular filtration rate, sodium output, urine excretion. Plasma levels of ANP in rats implanted with the PCL-tubes containing engineering cells is higher than that of the control rats. CONCLUSION: This study demonstrates encapsulated engineering cells have significant potential in treatment of hypertension. PMID- 12728967 TI - Erectile dysfunction after radical prostatectomy treatment and challenge. PMID- 12728968 TI - Optic pathway tumors in children. AB - Tumors of the optic pathways are sub-divided in this review into those that arise in one or both optic nerves anterior to the chiasm (optic nerve tumors); those that arise within the chiasm and do not extend significantly into the hypothalamus (optic chiasmatic tumors) and the large exophytic tumors that involve both the optic chiasm and the hypothalamus to a lesser or greater degree (optic chiasmatic/hypothalamic tumors). The management of optic chiasmatic gliomas is controversial, partly related to failure to separate out chiasmatic tumors from the chiasmatic/hypothalamic tumors. The optic nerve tumors are reviewed briefly, since they rarely extend intracranially. Chiasmatic tumors tend to be associated with NF1 and to behave almost like hamartomas. Close observation is usually the most appropriate management. On the other hand, chiasmatic/hypothalamic tumors grow like typical neoplasms. The tumors are almost uniformly low grade astrocytomas, but growth rates may berapid, especially in infants. Modern management has trended away from radical surgical resection, which has significant morbidity, to chemotherapy as the first line of treatment. In this review, the clinical presentation and management of different types of optic pathway tumors are discussed. PMID- 12728969 TI - The impact on sexual function after nerve sparing and non-nerve sparing radical retropubic prostatectomy. AB - BACKGROUND: We examine the impact of nerve sparing technique on the sexual function after radical retropubic prostatectomy for localized prostate cancer. METHODS: Between March, 1996 and September 2000, 44 men with clinically localized adenocarcinoma of the prostate who underwent radical retropubic prostatectomy (RRP) were included in this study. The mean age of these patients was 68.7 +/- 4.5 years old (range: 57-75). Among them, 8 patients were impotent preoperatively, 18 patients did not undergo nerve sparing surgery due to technical difficulties, and the remaining 18 potent patients underwent nerve sparing RRP. The postoperative sexual function was assessed by the International Index of Erectile Function (IIEF-6) and patient-reported percentage of recovery of sexual function. RESULTS: Mean length of follow-up was 14.4 +/- 2.7 months (range 12-18). The IIEF scores and % recovery of sexual function were significantly higher in nerve sparing surgery when compared to that of non-nerve sparing surgery (15.2 +/- 9.0 vs. 2.0 +/- 3.8 and 55 +/- 30% vs. 3 +/- 10%, respectively, p < 0.001). Of the 18 potent patients who underwent nerve sparing surgery, spontaneous erection and successful sexual activity was reported in 4 (22.2%) patients. Eight (44.4%) patients could achieve intercourse either with the aid of sildenafil or intracavernous alprostadil injection. Four (22.2%) patients had partial erection but refused further treatment. Two (11.1%) patients were completely impotent after nerve sparing surgery. Postoperative PSA was 0.10 +/- 0.18 (range 0.01-0.59). CONCLUSIONS: After a mean length of 14.4 months' follow-up, a majority of our patients could achieve sexual activity, and the cancer control following nerve sparing surgery was good. Our results suggest that nerve sparing retropubic radical prostatectomy is indicated in selected patients with localized adenocarcinoma of the prostate. PMID- 12728970 TI - Radiological manifestations of cryptococcal infection in central nervous system. AB - BACKGROUND: Cryptococcus neoformans is one of the most common infectious agents causing central nervous system (CNS) infections in immunocompromised patients. Patients with CNS cryptococcosis commonly present with non-specific manifestations such as headache, fever, seizure and conscious disturbance. This article presents the computerized tomography scans (CT) and magnetic resonance images (MRI) findings of cryptococcal infection in CNS and displays the image difference between immunocompromised and non-immunocompromised patients. METHODS: CNS cryptococcal infection was found in 26 cases over the past 14 years in Taipei Veterans General Hospital, including 18 males and 8 females. Ten patients were immunocompromised. In all cases, the CNS cryptococcosis was diagnosed by positive cryptococcal antigen titer and India ink stain from cerebral spinal fluid (CSF). Their clinical data and findings from CT and MRI were retrospectively studied. RESULTS: The most common CT manifestation is ventricular dilatation; other common findings include leptomeningeal enhancement, infarction (half of them revealed dilatation of Virchow-Robin spaces in subsequent MRI study) and normal appearance. Less common findings are ventriculitis and abscess formation. The most common MRI pattern is Virchow-Robin space dilatation and leptomeningeal enhancement. Other common appearance includes ventricular dilatation. Less common manifestations include cryptococcoma, hemorrhagic infarction and pontine abscess. There is no significant difference in image patterns between immunocompromised and non-immunocompromised patients (p > 0.05). CONCLUSIONS: The most common pattern ofCNS cryptococcosis is ventricular dilatation in CT and Virchow-Robin space dilatation in MRI. MRI is more sensitive in detecting CNS cryptococcal infection like Virchow-Robin space dilatation and leptomeningeal enhancement. There is no significant pattern difference between immunocompromised and non immunocompromised patients with CNS cryptococcosis. PMID- 12728971 TI - Esophageal manometry in patients with clinical symptoms mimicking esophageal origin: a hospital-based ten-year experience. AB - BACKGROUND: Primary esophageal motility disorder, which can cause chest pain or dysphagia, is seldom reported in Chinese. With the introduction of an easy and less uncomfortable method to perform esophageal manometry by low-compliance perfusion system, we studied symptomatic patients for more than 10 years. These data were analyzed and were compared to Western reports. METHODS: From August 1989 to June 1999, 264 patients with symptoms mimicking esophageal origin, such as chest pain, dysphagia or odynophagia, but without secondary motility disorders were enrolled. Esophageal manometry was performed on each patient. RESULTS: Among 264 manometric tracings, 142 (54%) were normal and 122 (46%) were abnormal. In patients with abnormal tracings, 73 were nonspecific esophageal motility disorder (NEMD), 20 were achalasia, 9 were diffuse esophageal spasm (DES), 8 were nutcracker esophagus, 7 were hypotensive low esophageal sphincter (LES), 3 were abnormal provocative test by edrophonium, and 2 were hypertensive LES. As in Western countries, the most common abnormality was NEMD. However, our series did not find many patients with DES, nutcracker esophagus and hypertensive LES. Similar results were noted in patients with NEMD, that most had increased nontransmitted contractions and low contraction amplitude. CONCLUSIONS: We found that primary esophageal motility disorder is not uncommon in Taiwan. Esophageal manometry should always be considered in patients with symptoms mimicking esophageal origin. PMID- 12728973 TI - Prevalence of antilipemic drug use in Taiwan: analysis of a sampling cohort within the national health insurance. AB - BACKGROUND: Hyperlipidemia is a major risk factor of cardiovascular diseases. We investigated the utilization ofantilipemic drugs at the outpatient sector within the National Health Insurance in Taiwan. METHODS: We obtained the first cohort (n = 50,000) dataset from the National Health Insurance Research Database and analyzed the outpatient claim files of the cohort in 2000. The antilipemic drugs were defined as the drug items belonging to the group C10 (serum lipid reducing agents) of the Anatomical Therapeutic Chemical classification system. RESULTS: Among the cohort with 46,614 eligible people, 760 patients had ever received antilipemic drugs (prevalence: 1.6%). The group 60-69 years of age had the greatest age-specific prevalence (7.2%), followed by the group over 70 years of age (6.0%). There were more male than female patients, but female patients outnumbered male patients before the age of 49 years. The antilipemic drugs had been prescribed 3,850 times totally with 70,272 defined daily doses (DDDs). On an average, a patient with antilipemic therapy received 5.1 (+/- 4.5) prescriptions of antilipemic drugs in one year and a prescription contained 18.3 (+/- 11.5) DDDs. We measured 4.1 DDDs per 1,000 inhabitants per day for all antilipemic drug use in 2000. The statins and fibrates predominated the antilipemic drug use. While gemfibrozil was most popular in respect of recipients and prescription items, simvastatin had the largest amount of use in unit of DDDs. Diabetes mellitus co-existed in 37.8% of the patients with antilipemic therapy and the standardized morbidity ratio (SMR) was 3.34. The other concomitant diseases included essential hypertension (rate: 48.8%, SMR: 2.40) and other heart disease (rate: 30.7%, SMR: 2.36). CONCLUSIONS: Statins were the leading antilipemic drugs in Taiwan. The users of antilipemic drugs were more likely to have concomitant diabetes mellitus, hypertension and heart disease. PMID- 12728972 TI - Sclerosing hemangioma of the lung: an analysis of 44 cases. AB - BACKGROUND: Sclerosing hemangioma is a rare benign lung tumor. Preoperative diagnosis of this tumor is difficult, and sometimes even intraoperative frozen sections cannot differentiate it from malignant tumors. Herein, we present our experiences in investigating its characteristics. METHODS: The medical records and pathological examinations of patients diagnosed with sclerosing hemangioma from 1982 to 2001 at the Division of Thoracic Surgery in Taipei-VGH were retrospectively reviewed. RESULTS: The incidence of sclerosing hemangioma in benign lung tumors resected during that period was 32.8%. There were 44 patients (7 male and 37 female) aged from 16 to 72 years, with a peak at fifth decade, and 72.7% of them were asymptomatic. Accurate preoperative diagnosis by chest CT could be achieved in only 20% (4/20) of patients, and malignancy could not be ruled out in 40% (8/20) of tumors. Histologically, this tumor exhibits four major patterns: solid, sclerotic, papillary and hemorrhagic. Five patients in this series had predominantly one pattern. At least two patterns existed in the remaining 39 patients, including eight patients who had tumors with all four patterns. There was no operative mortality or tumor recurrence despite that different operative methods were undertaken. CONCLUSIONS: Although rare, sclerosing hemangioma still accounts for the second common benign lung tumor in the current series. Chest CT and bronchoscopic examinations could not make accurate diagnosis preoperatively, and thoracotomy is usually indicated for definite diagnosis and treatment. Limited resection is warranted in view of uncommon tumor recurrence. In addition, multiple nodular lesions could not exclude the possibility of sclerosing hemangioma. PMID- 12728974 TI - Endogenous uveitis: experiences in Kaohsiung Veterans General Hospital. AB - BACKGROUND: The distribution of uveitis varies in different parts of the world, which is attributed to geographic, alimentary or genetic factors. Epidemiological studies in uveitis may be extremely important in an effort to better understand the etiology of the disease and the classification of its subtypes. The purpose of this study was to determine the profile and characteristics of endogenous uveitis in southern Taiwan. METHODS: Retrospective chart review ofthe patients diagnosed and treated as endogenous uveitis in Kaohsiung Veterans General Hospital from January 1991 to December 2000 was performed. Demographic data, past history, ophthalmic examination and other laboratory tests were recorded and analyzed. RESULTS: One-hundred and sixty patients were collected, including 93 (58.1%) males and 67 (41.9%) female, with average age of 41.1 +/- 17.1 years (range 16-77), and mean follow-up period of 13.7 months. The diagnoses included 46 (28.8%) cases of acute anterior nongranulomatous uveitis (AAU), 26 (16.3%) cases of Vogt-Koyanagi-Harada (VKH) disease, 18 (11.3%) cases of intermediate uveitis, 14 (8.8%) cases of Behcet's disease, 26 (16.3%) cases of infectious origin, 7 (4.4%) cases of other diagnoses, and 23 (14.4%) undefined cases. Final visual acuity equal to or better than 0.5 was obtained in 78.2% of eyes of AAU, 65.4% of eyes of VKH disease, 48.1% of eyes of Behcet's disease, and 66.7% of eyes of intermediate uveitis. CONCLUSIONS: Acute anterior nongranulomatous uveitis was the most common endogenous uveitis, while VKH was the most common panuveitis in this series. As infectious origin contributes a significant proportion of endogenous uveitis, infectious etiology should be considered in any type of uveitis that responds poorly to conventional treatment. PMID- 12728975 TI - Study of the polymorphism of angiotensinogen, anigiotensin-converting enzyme and angiotensin receptor in type II diabetes with end-stage renal disease in Taiwan. AB - BACKGROUND: This study compared the polymorphisms of angiotensin-converting enzyme (ACE), angiotensinogen (AGT) and angiotensin II type 1 receptor (AT1R) gene between type II diabetes with diabetic nephropathy (DN) in end-stage renal disease (ESRD) and those of the normal individuals in Taiwan. METHODS: 129 patients with type II diabetes in ESRD from three dialysis centers were compared to 116 age and blood pressure-matched normal individuals. Polymerase chain reaction (PCR) specific for ACE gene was tested for insertion/deletion polymorphism. M235T and T174M variant polymorphisms of the AGT gene and the AT1R A1166C polymorphism were also tested according to previously described protocols. RESULTS: Between the two groups, frequencies of the alleles and the genotypes in ACE gene polymorphism were comparable. There was no significant difference of the ACE insertion and deletion polymorphism and AT1R gene polymorphism between the two groups. The allele frequencies of AGT point mutation at 235 (M235T) was significantly higher in DM nephropathy but the genotypic frequencies were not. In the AGT point mutation at 174 (T174M), the frequencies of both of alleles and genotypes were significantly higher in the DM nephropathy patients. CONCLUSIONS: In conclusion, there were significantly higher frequencies of alleles and genotypes in AGT point mutation at 174, which had never been reported before. In AGT point mutation at 235, there were significantly higher frequencies of alleles, but not genotypes in diabetic nephropathy. Our study suggested that the AGT gene plays an important role in diabetic nephropathy. PMID- 12728976 TI - Dose-response relationships of propranolol in Chinese subjects with different CYP2D6 genotypes. AB - BACKGROUND: For clinical treatment, a smaller dosage of propranolol is often used among Chinese people. Propranolol is metabolized by polymorphic CYP2D6. We postulate that the lower propranolol dosage in Chinese is due to a slower CYP2D6 metabolism. A majority of the Chinese population has the nucleotide T188 in the CYP2D6 gene (CYP2D6*10) instead of C188 (CYP2D6*1), which most white subjects have. Chinese subjects of different CYP2D6*1/CYP2D6*10 genotypes have been shown to have different propranolol pharmacokinetic characteristics. In this study, we compared the beta-blockade effects of propranolol in Chinese subjects of the two different CYP2D6 genotypes. METHODS: Based on the nucleotide 188 genotypes, two groups of 10 healthy subjects each were selected. Each subject was given a 10-, 20-, or 40-mg rac-propranolol tablet three times a day for 3 days in 3 different phases. Heart rate and blood pressure were measured in both supine and upright positions. The heart rate was also determined during treadmill exercise test. Plasma concentration of S-propranolol at 2 hrs after the last-dose administration was measured. RESULTS: Despite therebeing higher S-propranolol plasma concentration in CYP2D6*10 subjects than in CYP2D6*1 subjects at 10- and 20-mg dosage, the dose-response relationship was not significantly different in these subjects. CONCLUSIONS: Our results do not support the hypothesis that CYP2D6*1/CYP2D6*10 polymorphism may affect the beta-blockade effect of propranolol in Chinese subjects. PMID- 12728977 TI - Hypertrophic pulmonary osteoarthropathy associated with disease progression in renal cell carcinoma. AB - Hypertrophic osteoarthropathy (HOA) is a paraneoplastic syndrome characterized by periosteal formation and arthritis and usually accompanied by clubbing ofthe digits. Many malignancies have been associated with HOA/clubbing, most being lung cancer and lung metastatic cancer. We herein present a 53-year-old man with lung metastasis from renal cell carcinoma (RCC). HOA occurred one year after the metastasis. Reviewing the literature, only five cases of RCC with HOA have been reported. If their clinical history was traceable, they consistently had disease progression. We reviewed the pathogenesis of HOA/clubbing and linked the prognosis of RCC to relevant cytokines. Therefore, HOA not only heralds a progression of disease but suggests a probable therapeutic choice by targeting some cytokines. PMID- 12728978 TI - Primary adrenal lymphoma manifestating as adrenal incidentaloma. AB - Although involvement of the adrenals by malignant lymphoma is common, primary adrenal lymphoma is extremely rare. Herein, we report a case of a 59-year-old woman with bilateral adrenal glands enlargement found incidentally on abdominal imaging. Despite of the huge size of the tumor, her adrenal function was intact. Ultrasound-guided biopsy disclosed a picture of malignant lymphoma, diffuse large B cell type. The patient received bilateral adrenalectomy and adjuvant chemotherapy, but succumbed 6 months later. We suggest that, although rare, primary adrenal lymphoma should be considered as a possible cause of bilateral adrenal incidentalomas. PMID- 12728979 TI - Transplantation of related histocompatible marrow and peripheral blood stem cells in a patient with severe combined immunodeficiency and disseminated BCG infection. AB - We report the results of allogeneic bone marrow transplantation (BMT) in a 15 month-old boy with severe combined immunodeficiency (SCID) complicated by disseminated Bacillus Calmette-Guerin (BCG) infection. Slow immunological recovery after BMT is considered to be associated with prolonged BCG infection. After one booster dose of allogeneic peripheral blood stem cell transplantation (PBSCT), there was still little improvement in immune reconstitution. The patient finally expired because of failure to achieve a functioning immune system after BMT and septic shock. PMID- 12728980 TI - Genetics of polycyclic aromatic hydrocarbon metabolism in diverse aerobic bacteria. AB - Polycyclic aromatic hydrocarbons (PAHs), which consist of two or more fused aromatic rings, are widespread in the environment and persist over long periods of time. The decontamination of a PAH-polluted environment is of importance because some PAHs are toxic, mutagenic, and carcinogenic and therefore are health hazards. As part of the efforts to establish remediation processes, the use of aerobic bacteria has been extensively studied, and both enzymologic and genetic studies are underway for the purpose of effective biodegradation. In the last two decades, one highly conserved group of PAH-catabolic genes from Pseudomonas species, called the nah-like genes, has been well investigated, and much has been found, including the structure-function relationships and the evolutionary trails of the catabolic enzymes. However, recently, PAH-catabolic genes, which are evolutionarily different from the nah-like genes, have been characterized from both Gram-negative bacteria other than Pseudomonas species and Gram-positive bacteria, and the information about these genes is expanding. This review is an outline of genetic knowledge about bacterial PAH catabolism. PMID- 12728981 TI - Effects of corticosterone on Ca2+ uptake and myofibrillar disassembly in primary muscle cell culture. AB - This study was done to examine the effects of corticosterone, a glucocorticoid, on Ca2+ uptake, proteolysis, and Ca2+ channels in primary cultures of chick muscle cells, to clarify the mechanism of glucocorticoid action on muscle proteolysis. Chick muscle cells were incubated for 24 h in a medium containing corticosterone (30 ng/ml) when the cells were confluent (6 days). To examine the contribution of Ca2+ channels, nifedipine, a Ca2+ channels antagonist, was used. Ca2+ uptake measured with 45CaCl2 was increased three-fold by corticosterone, with a peak at 12 h after the treatment started. The growth of the cells estimated from the protein content and creatine kinase activity was not affected by corticosterone. Proteolysis, evaluated with [3H]tyrosine as a label of the protein and Ntau-methylhistidine release, was unchanged by corticosterone. However, the amount of easily releasable myofilament as a measure of myofibrillar disassembly in the muscle cells was increased by corticosterone, and prevented by nifedipine. These results show that corticosterone increases Ca2+ uptake and starts myofibrillar protein breakdown. PMID- 12728982 TI - Role of endo-1,4-beta-glucanases from neisseria sicca SB in synergistic degradation of cellulose acetate. AB - An enzyme hydrolyzing beta-1,4 bonds in cellulose acetate was purified 10.5-fold to electrophoretic homogeneity from a culture supernatant of Neisseria sicca SB, which assimilate cellulose acetate as the sole carbon and energy source. The enzyme was an endo-1,4-beta-glucanase, to judge from the substrate specificity and hydrolysis products of cellooligosaccharides, we named it endo-1,4-beta glucanase I (EG I). Its molecular mass was 50 kDa, 9 kDa larger than EG II from this strain, and its isoelectric point was 5.0. Results of N-terminal and inner peptide sequences of both enzymes, and a similarity search, suggested that EG I contained a carbohydrate-binding module at the N-terminus and that EG II lacked this module. The pH and temperature optima of EG I were 5.0-6.0 and 45 degrees C. It hydrolyzed water-soluble cellulose acetate (degree of substitution, 0.88) and carboxymethyl cellulose. The Km and Vmax for these compounds were 0.296% and 1.29 micromol min(-1) mg(-1), and 0.448% and 13.6 micromol min(-1) mg(-1), respectively. Both glucanases and cellulose acetate esterase from this strain degraded water-insoluble cellulose acetate synergistically. PMID- 12728983 TI - Enzymatically depolymerized alginate oligomers that cause cytotoxic cytokine production in human mononuclear cells. AB - Enzymatically depolymerized guluronate and mannuronate oligomers were prepared from polyuronates with an alginate lyase from a Pseudoalteromonas sp., and their effects on mononuclear cells from human peripheral blood were examined. Conditioned medium prepared by the incubation of cells with an untreated polyuronate had little effect on growth of human leukemic U937 cells, but a medium prepared with depolymerized uronate oligomers inhibited their growth. Inhibition was greater in a medium prepared with guluronate oligomer than one prepared with mannuronate oligomer. The cytotoxic activity of the medium was heat labile and nondialyzable. Apoptotic nuclear morphological changes and increased caspase-3-like activity were found in U937 cells treated with a medium prepared with depolymerized uronates. The medium prepared with purified tetra-guluronate and tetra-mannuronate also was cytotoxic; these effects were inhibited by antibodies to tumor necrosis factor-alpha. Our results suggested that enzymatically depolymerized guluronate and mannuronate oligomers induced the production of cytotoxic cytokines in human mononuclear cells, although the uronate polymers before depolymerization had no such activity. PMID- 12728985 TI - Wound-responsive cis-element in the 5'-upstream region of cucumber ascorbate oxidase gene. AB - The cucumber (Cucumis sativas) AAO1 gene (former name, Aso1) encodes an ascorbate oxidase that catalyzes the oxidation by molecular oxygen of ascorbic acid to dehydroascorbate. CsAAO1 mRNA concentrations rose rapidly after mechanical wounding of cucumbers. To study the wound-responsive expression of CsAAO1 in detail, we examined transgenic tobacco plants harboring a CsAAO1 promoter-beta glucuronidase fusion gene. CsAAO1 promoter activity in leaves of the tobacco was induced by wounding. Analysis of the regulatory properties of 5'-deleted promoter fragments showed that a putative wound-responsive cis-element (WRE) was located 736 to -707 bp from the translation initiation site. DNA binding factors that bound specifically to the putative WRE sequence were identified in tobacco nuclear extracts by gel retardation assays. PMID- 12728984 TI - Substrate specificities of deuterolysin from Aspergillus oryzae and electron paramagnetic resonance measurement of cobalt-substituted deuterolysin. AB - The substrate specificities of deuterolysin, a 19-kDa zinc-protease (EC 3.4.24.39) from Aspergillus oryzae, were investigated at pH 9.0 with various fluorogenic acyl-peptide-4-methylcoumaryl-7-amides (peptide-MCAs). N Butoxycarbonyl-Arg-Val-Arg-Arg-MCA was the best substrate for deuterolysin. We therefore measured its kinetic parameters. Deuterolysin had high activity toward the peptide bonds next to pairs of basic residues in calf thymus histone H4. The specificity of cobalt-substituted deuterolysin (Co-deuterolysin) for peptide-MCAs was similar to that of native deuterolysin. The CD spectrum of Co-deuterolysin was similar to that of the native deuterolysin. The metal coordination sphere of Co-deuterolysin was analyzed by Q-band (33.9570 GHz) electron paramagnetic resonance (EPR) spectroscopy. Using computer simulation of EPR, we found the g principal values to be g(xx) = 5.20, g(yy) = 4.75, and g(zz) = 2.24; the metal center was a divalent cobalt ion in a high spin state. PMID- 12728987 TI - Fermentation conditions and properties of a chitosanase from Acinetobacter sp. C 17. AB - A species of bacterium with high chitosanase activity was isolated from soil samples in Haiyan City, China, and identified as an Acinetobacter species. This strain, named Acinetobacter sp. strain C-17, produced a chitosanase that was inducible and secreted into the medium. The optimal conditions for enzyme production were cells used to inoculate a medium containing 1% chitosan (pH 7.0) followed by culture at 30 degrees C. The chitosanase activity reached 1.7 U/ml when strain C-17 was incubated in a 250-ml flask under the optimal conditions for 24 h, and reached 2.8 U/ml when cells were incubated in a 3-l fermentor. The optimal pH and temperature for hydrolysis of chitosanase were 7.0 and 36 degrees C, respectively. The chitosanase activity was stable in the pH range of 5-8 and temperature range of 30-40 degrees C. The chitosanase of the strain was extracted by zinc acetate and ammonium sulfate precipitation. The molecular mass was estimated to be 35.4 kDa by SDS-PAGE. PMID- 12728986 TI - Reduction of paraquat-induced oxidative stress in rats by dietary soy peptide. AB - The effect of a dietary soy protein isolate (SPI), soy peptide (PEP) and the amino acids in soy protein on paraquat (PQ)-induced oxidative stress was investigated in rats. In the first experiment, male Wistar rats were fed on experimental diets containing casein (CAS), SPI and PEP as nitrogen sources with or without 0.025% PQ. The reduced food intake and body weight gain of the rats fed with PQ was mitigated by either the SPI or PEP intake. Both SPI and PEP prevented the elevation of the serum TBARS concentration and tended to prevent the elevation of lung weight induced by PQ. In the second experiment, the rats were fed on diets containing an amino acid mixture resembling casein (CASAA) or soy protein (SPIAA) with or without PQ. The SPIAA intake did not affect the reduction of food intake and body weight gain, nor the elevation of lung weight and TBARS in the serum and liver induced by PQ. These results demonstrate that the intake of either dietary SPI or PEP, but not an amino acid mixture resembling soy protein, had the effect of reducing PQ-induced oxidative stress in rats. PMID- 12728988 TI - Structure of folding intermediates at pH 4.0 and native state of microbial transglutaminase. AB - Recombinant microbial transglutaminase has been expressed in Escherichia coli as insoluble inclusion bodies. After we searched for refolding conditions, refolding of the protein could be done by first dilution of the unfolded enzyme in a buffer at pH 4.0, and then by titration of the pH from 4.0 to 6.0. CD analysis showed that a burst of secondary structure formation occurred within the dead time of the experiment and accounted for 75% of the signal change in the far UV CD, with little tertiary structure being formed. This burst was followed by slow rearrangement of the secondary structure accompanied by formation of tertiary structure. The secondary and tertiary structures of the final sample at pH 4.0, corresponding to the folding intermediate, were different from these structures at pH 6.0. Once the native structure was obtained, acidification of the native protein to pH 4.0 did not lead to a structure like that of the folding intermediate. Sedimentation velocity analysis showed that the folding intermediate had an expanded structure and contained no other structure species including large aggregates. PMID- 12728989 TI - Reduction mechanism of tetrazolium salt XTT by a glucosamine derivative. AB - XTT (3'-[1-[(phenylamino)-carbonyl]-3,4-tetrazolium]-bis(methoxy-6 nitro)benzenesulfonic acid hydrate) was reduced by incubated glucosamine hydrochloride. The XTT reducibility by incubated glucosamine was linearly related with the DNA-breaking activity. In order to elucidate the reaction mechanism, the glucosamine derivatives formed during the incubation process were separated by HPLC, and the compound responsible for the reduction was analyzed. Among the incubated products, fructosazine and deoxyfructosazine were identified by LC-MS, FAB-MS, and 1H- and 13C-NMR. These products showed no XTT reducibility, but an unstable intermediate with a molecular weight of 322 displayed reducibility. Since the intermediate gave fructosazine by oxidation with XTT and was a precursor of deoxyfructosazine, we conclude that the intermediate could have been dihydrofructosazine. Therefore, the XTT reducibility by incubated glucosamine was based on dihydrofructosazine formed by the condensation of two molecules of glucosamine. PMID- 12728990 TI - Expression, purification, and characterization of 2'-aminobiphenyl-2,3-diol 1,2 dioxygenase from carbazole-degrader Pseudomonas resinovorans strain CA10. AB - The two-subunit meta-cleavage enzyme, 2'-aminobiphenyl-2,3-diol 1,2-dioxygenase (CarBaBb), from the carbazole degrader Pseudomonas resinovorans strain CA10 was purified to homogeneity from an Escherichia coli strain carrying the expression vector pUCA503, in which two copies of the carBaBb genes are tandemly linked. SDS PAGE and gel filtration showed that CarB was a alpha2beta2-heterotetrameric enzyme with subunit molecular masses of approximately 10,000 for CarBa and 29,000 for CarBb. The optimum pH for activity was 8.5 and that of temperature was 35 degrees C. The CarB enzyme had a Km of 14 microM and a kcat/Km of 0.25 microM(-1) s(-1) for 2'-aminobiphenyl-2,3-diol, and the catalytic activities for biphenyl type catecholic substrates were higher than those for monoaromatic catechol derivatives. The enzyme was originally isolated as a meta-cleavage enzyme for 2' aminobiphenyl-2,3-diol involved in carbazole degradation, but the enzyme was highly specific for 2,3-dihydroxybiphenyl. PMID- 12728991 TI - (E)-2-(2-Hydroxyethylidene)-6-methyl-5-heptenal (alpha-acariolal) and (E)-2-(2 hydroxyethyl)-6-methyl-2,5-heptadienal (beta-acariolal), two new types of isomeric monoterpenes from Caloglyphus polyphyllae (Acari: Acaridae). AB - The opisthonotal gland secretion of the acarid mite, Caloglyphus polyphyllae, contained two new monoterpenes, (E)-2-(2-hydroxyethylidene)-6-methyl-5-heptenal (1) and (E)-2-(2-hydroxyethyl)-6-methyl-2,5-heptadienal (2), to which we have given the trivial names alpha- and beta-acariolal in relation to alpha- and beta acaridial (3 and 4), respectively. Elucidation of the structure of 1 was established mainly from 1H-NMR and GC/MS spectral data after partial purification, together with the fact that 1 was recovered in the more-polar fraction from a silica gel column than alpha- and beta-acaridial (3 and 4) present in the secretion. Compound 2 was obtained in the same fraction as a mixture with 1. Based on the facts that 2 had the same molecular weight by GC/MS and the same polarity as that of 1, compound 2 was assumed to be a structural analog of 1. The structures of compounds 1 and 2 were confirmed by their synthesis in nine and ten respective steps starting from alpha-bromo-gamma butyrolactone. PMID- 12728992 TI - Insulin-dependent adipogenesis in stromal ST2 cells derived from murine bone marrow. AB - ST2 cells, a cloned stromal-cell line from mouse bone marrow have the phenotypes of osteoblasts and hematopoietic supporting cells, but little is known about the adipogenesis in this cell line. We found that treatment of ST2 cells with a cocktail, a combination of insulin, dexamethasone (DEX) and 3-isobutyl-1 methylxanthine (IBMX), induced adipocyte differentiation. The cells were adipocytic, as seen by the accumulation of triglycerides and with lipid droplets stained with Oil Red O. Expressions of the adipogenic transcriptional factors peroxisome-proliferator-activated receptor gamma and CCAAT-enhancer binding protein alpha were stimulated, triggering the expression of the late adipocyte specific marker alpha-glycerophosphate-3-dehydrogenase. Unlike another bone marrow stromal cell line PA6, ST2 cells responded to the adipogenic effects of insulin, as do the extramedullary preadipocytes 3T3-L1. Like PA6 and 3T3-L1 cells, adipogenesis in ST2 cells was inhibited by 1,25-dihydroxyvitamin D3, retinoic acid, tumour necrosis factor alpha, and transforming growth factor beta. Therefore, ST2 cells can differentiate into osteoblasts, adipocytes, and hematopoietic supporting cells, in which the process of adipogenesis differs from that of bone marrow preadipocytes PA6, and resembles that of the extramedullary preadipocytes 3T3-L1. PMID- 12728993 TI - N-cyanomethyl-2-chloroisonicotinamide induces systemic acquired resistance in arabidopsis without salicylic acid accumulation. AB - Systemic acquired resistance (SAR) is a potent innate immunity system in plants that is induced through the salicylic acid-mediated pathway. N-cyanomethyl-2 chloroisonicotinamide (NCI) is able to induce a broad range of disease resistance in tobacco and rice and induces SAR marker gene expression without SA accumulation in tobacco. To clarify the detailed mode of action of NCI, we analyzed its ability to induce defense gene expression and resistance in Arabidopsis mutants that are defective in various defense signaling pathways. Wild-type Arabidopsis treated with NCI exhibited increased expression of several pathogenesis-related genes and enhanced resistance to the bacterial pathogen, Pseudomonas syringae pv. tomato DC3000. NCI induced disease resistance and PR gene expression in NahG transgenic plants, but not in the npr1 mutant. NCI could induce PR gene expression in the etr1-1, ein2-1 and jar1-1 mutants. Thus, NCI activates SAR, independently from ethylene and jasmonic acid, by stimulating the site between SA and NPR1. PMID- 12728994 TI - Determination of the absolute configuration of (+)-xestoaminol C [(2S, 3R)-2 amino-3-tetradecanol], a metabolite of Fiji sponge, Xestospongia sp., by the synthesis of its N,O-diacetyl derivative. AB - The N,O-diacetyl derivative of (+)-xestoaminol C (2-amino-3-tetradecanol, 1), an inhibitor of reverse transcriptase isolated from Xestospongia sp., was synthesized from (S)-alanine, and its absolute configuration was determined as 2S, 3R. PMID- 12728995 TI - Isolation of Paenibacillus illinoisensis that produces cyclodextrin glucanotransferase resistant to organic solvents. AB - A bacterium that secreted cyclodextrin glucanotransferase (CGTase) in a medium overlaid with n-hexane was isolated and identified as Paenibacillus illinoisensis strain ST-12 K. The CGTase of the strain was purified from the culture supernatant. The molecular mass was 70 kDa. The enzyme was stable at pH 6 to 10 and active at pH 5.0 to 8.0. The optimum temperature at pH 7.0 was 65 degrees C in the presence of 5 mM CaCl2. The enzyme produced mainly beta-cyclodextrin. The total yield of alpha-, beta-, and gamma- cyclodextrins was increased 1.4-fold by the addition of ethanol. In particular, the yield of beta-cyclodextrins in the presence of 10% (vol/vol) ethanol was 1.6-fold that without ethanol. The CGTase was stable and active in the presence of large amounts of various organic solvents. PMID- 12728996 TI - Mutational analysis of amino acid residues involved in catalytic activity of a family 18 chitinase from tulip bulbs. AB - We expressed chitinase-1 (TBC-1) from tulip bulbs (Tulipa bakeri) in E. coli cells and used site-directed mutagenesis to identify amino acid residues essential for catalytic activity. Mutations at Glu-125 and Trp-251 completely abolished enzyme activity, and activity decreased with mutations at Asp-123 and Trp-172 when glycolchitin was the substrate. Activity changed with the mutations of Trp-251 to one of several amino acids with side-chains of little hydrophobicity, suggesting that hydrophobic interaction of Trp-251 is important for the activity. Molecular dynamics (MD) simulation analysis with hevamine as the model compound showed that the distance between Asp-123 and Glu-125 was extended by mutation of Trp-251. Kinetic studies of Trp-251-mutated chitinases confirmed these various phenomena. The results suggested that Glu-125 and Trp-251 are essential for enzyme activity and that Trp-251 had a direct role in ligand binding. PMID- 12728997 TI - Novel gene encoding a Ca2+-binding protein and under hexokinase-dependent sugar regulation. AB - A cDNA encoding a predicted 15-kDa protein was earlier isolated from sugar induced genes in rice embryos (Oryza sativa L.) by cDNA microarray analysis. Here we report that this cDNA encodes a novel Ca2+-binding protein, named OsSUR1 (for Oryza sativa sugar-up-regulated-1). The recombinant OsSUR1 protein expressed in Escherichia coli had 45Ca2+-binding activity. Northern analysis showed that the OsSUR1 gene was expressed mainly in the internodes of mature plants and in embryos at an early stage of germination. Expression of the OsSUR1 gene was induced by sugars that could serve as substrates of hexokinase, but expression was not repressed by Ca2+ signaling inhibitors, calmodulin antagonists and inhibitors of protein kinase or protein phosphatase. These results suggested that Os-SUR1 gene expression was stimulated by a hexokinase-dependent pathway not mediated by Ca2+. PMID- 12728998 TI - Novel chitosanase from Streptomyces griseus HUT 6037 with transglycosylation activity. AB - Streptomyces griseus HUT 6037 inducibly produced two chitosanases when grown on chitosan. To elucidate the mechanism of degradation of chitinous compound by this strain, chitosanases I and II of S. griseus HUT 6037 were purified and characterized. The purified enzymes had a molecular mass of 34 kDa. Their optimum pH was 5.7, and their optimum temperature was 60 degrees C. They hydrolyzed not only partially deacetylated chitosan, but also carboxymethylcellulose. Time dependent 1H-NMR spectra showing hydrolysis of (GlcN)6 by the chitosanases were obtained for identification of the anomeric form of the reaction products. Both chitosanases produced the beta-form specifically, indicating that they were retaining enzymes. These enzymes catalyzed a glycosyltransfer reaction in the hydrolysis of chitooligosaccharides. The N-terminal and internal amino acid sequences of chitosanase II were identified. A PCR fragment corresponding to these amino acid sequences was used to screen a genomic library for the entire gene encoding chitosanase II. Sequencing of the choII gene showed an open reading frame encoding a protein with 359 amino acid residues. The deduced primary structure was similar to endoglucanase E-5 of Thermomonospora fusca, which enzyme belongs to family 5 of the glycosyl hydrolases. This is the first report of a family 5 chitosanase with transglycosylation activity. PMID- 12728999 TI - Effects of degree of hydrolysis and pH on the solubility of heme-iron enriched peptide in hemoglobin hydrolysate. AB - Hemoglobin was hydrolyzed with Esperase and Flavourzyme as the endopeptidase and exopeptidase, respectively. The solubility of the heme-iron enriched peptide fraction decreased as the degree of hydrolysis of the hydrolysate increased. When the pH of a hydrolysate was adjusted to 5.0 after simultaneous hydrolysis with the two enzymes, the solubility of heme-iron enriched peptide was nearly zero, and 98% of the heme-iron enriched peptide fraction was recovered as a precipitate. These results indicated that an effective separation method for the production of heme-iron enriched peptide could be established by pH adjustment of the hemoglobin hydrolysate with high degree of hydrolysis. PMID- 12729000 TI - Periplasmic secretion of native ovalbumin without signal cleavage in Escherichia coli. AB - In Escherichia coli cells carrying wild-type ovalbumin cDNA, some of the recombinant protein was secreted into the periplasmic space. In contrast, a signal-region mutant form of ovalbumin (deletion, Gly1 to Ala39) was not detected in the periplasm despite being synthesized at the same level as the wild-type protein. Chemical and spectroscopic analyses showed that periplasmic ovalbumin assumes a conformation indistinguishable from that of native egg white ovalbumin. We concluded that a process resembling the secretion of ovalbumin process in the oviduct occurs also in bacteria. PMID- 12729001 TI - Effect of a selected amino acid mixture on the recovery from muscle fatigue during and after eccentric contraction exercise training. AB - The effect of an amino acid mixture on the recovery from muscle fatigue after eccentric exercise (ECEX) training was examined in twenty-two male college students. The administration of 5.6 g of the amino acid mixture twice daily resulted faster recovery of muscle strength than that with a placebo. The oral administration of the amino acid mixture was proved to effective for muscle strength recovery after the eccentric exercise. PMID- 12729002 TI - New phenolic compounds from Kokuto, non-centrifuged cane sugar. AB - Five new phenolic compounds, 4-(beta-D-glucopyranosyloxy)-3,5-dimethoxyphenyl propanone (8), 3-[5-[(threo) 2,3-dihydro-2-(4-hydroxy-3-methoxyphenyl)-3 hydroxymethyl-7-methoxybenzofuranyl]-propanoic acid (12), 2-[4-(3-hydroxy-1 propenyl)-2,6-dimethoxyphenoxy]-3-hydroxy-3-(4-hydroxy-3,5-dimethoxyphenyl)propyl beta-D-glucopyranoside (13), 4-[(erythro) 2,3-dihydro-3(hydroxymethyl)-5-(3 hydropropyl)-7-methoxy-2-benzofuranyl]-2,6-dimethoxyphenyl-beta-D-glucopyranoside (14), 9-O-beta-D-xylopyranoside of icariol A2 (15), and known phenolic compounds were isolated from Kokuto, non-centrifuged cane sugar (Saccharum officinarum L.). Their structures were determined by a spectral investigation. PMID- 12729004 TI - Response of the induction of rat liver serine dehydratase to changes in the dietary protein requirement. AB - Growing and mature rats were examined for the effect of a change in dietary protein requirements on the induction of liver serine dehydratase (SDH). The rats were fed on diets varying in casein content, and the weight change and nitrogen balance was determined. SDH activity and its gene expression were induced in both growing and mature rats when their protein intake exceeded their nutritional requirements. PMID- 12729003 TI - Prolyl endopeptidase inhibitory peptides in wine. AB - Two peptides that inhibit prolyl endopeptidase were isolated from a red wine made from Cabernet Sauvignon grapes. Their amino acid sequences and IC50 values were Val-Glu-Ile-Pro-Glu (17.0 microM) and Tyr-Pro-Ile-Pro-Phe (87.8 microM). The peptides also suppressed the degradation levels of the bioactive peptides vasopressin, substance P, and neurotensin fragments 8-13, which are involved in memory and neural communication. PMID- 12729005 TI - Presence of free D-amino acids in microalgae. AB - Several species of microalgae (phytoplankton), 4 species of freshwater algae and 4 species of marine diatoms, were cultured germ-free in the laboratory. The presence of free D-amino acids was verified in these species by a reversed-phase HPLC analysis. D-Aspartate was detected in all the microalgae examined, but D alanine was only present in the marine diatoms. The D-amino acid content in Asterionella sp. of the marine diatoms increased from the exponential phase to the stationary phase and then decreased to the phase of decline. PMID- 12729006 TI - High IgM production by human-human hybridoma HB4C5 cells cultured in microtubes. AB - HB4C5 cells, a human-human hybridoma, were cultured in microtubes. After 24 h of cultivation, growth of the cells cultured in microtubes was 1.2- to 1.5-fold that in culture dishes or 96-well culture plates. The production of IgM was 2.6- to 3.3-fold that in the 96-well culture plates. PMID- 12729007 TI - Evaluation of the anti-oxidative effect (in vitro) of tea polyphenols. AB - Forty-three polyphenols from tea leaves were evaluated for their anti-oxidative effect against lipid peroxidation by the ferric thiocyanate method in vitro. Among these, 1,4,6-tri-O-galloyl-beta-D-glucose (hydrolyzable tannin) showed the highest anti-oxidative activity against lipid peroxidation, even stronger than that of 3-tert.-butyl-4-hydroxyanisole (BHA). The assay demonstrates that tea polyphenols, except for desgalloylated dimeric proanthocyanidins that possess a catechin structure in the upper unit and desgalloylated flavan-3-ols, and excepting theaflavin 3,3'-di-O-gallate, had more anti-oxidative activity than that of alpha-tocopherol. The chemical structure-activity relationship shows that the anti-oxidative action advanced with the condensation of two molecules of flavan-3-ols as well as with 3-O-acylation in the flavan skeleton such as that by galloyl, (3'-O-methyl)-galloyl, and p-coumaroyl groups. PMID- 12729008 TI - Effect of alpha-tocopherol on the oxidative modification of apolipoprotein E in human very-low-density lipoprotein. AB - The oxidative modification of apolipoprotein (apo) E and lipid peroxidation in human very-low-density lipoprotein (VLDL) induced by peroxynitrite and cupric ions in vitro were strongly suppressed by enrichment with alpha-tocopherol (alpha Toc; 170 microM). Alpha-Toc also suppressed the decrease in the heparin-binding activity of apoE in the VLDL oxidation. These results suggest that alpha-Toc protected apoE in VLDL from oxidative stress. PMID- 12729009 TI - Isothermal titration calorimetric studies on the binding of a family 6 carbohydrate-binding module of Clostridium thermocellum xynA with xlylooligosaccharides. AB - The family 6 carbohydrate-binding module (CBM) of Clostridium thermocellum XynA was expressed, and the binding equilibria of the CBM with xylooligosaccharides (degree of polymerization DP = 2-8) were observed by isothermal titration calorimetry (ITC) at pH 8. The association constant, Ka, increased with increasing DP from 5 x 10(3) M(-1) (DP = 2) to approximately 5 x 10(5) M(-1) (DP = 5-8) at 20 degrees C. The Ka values at 60 degrees C were about 1/10 of those at 20 degrees C. The binding was found to be an enthalpy-driven reaction. The DP dependence of the thermodynamic parameters of the binding reaction suggested the size of the ligand-binding site to be 5 xylose units long. PMID- 12729010 TI - Antioxidative compounds from Crotalaria sessiliflora. AB - Seven antioxidative compounds were isolated from the EtOAc extract of the aerial part of C. sessiliflora (Japanese name, tanukimame) by activity-guided fractionation with 2,2-diphenyl-1-picrylhydrazyl (DPPH). Among the isolated compounds, hydroxyeucomic acid showed the strongest free radical-scavenging activity, which was almost identical to that of epigallocatechin gallate, against DPPH. Orientin and isoorientin showed strong anti-peroxidative activities toward linoleic acid and protective effects against the bactericidal action of the tert butyl peroxyl radical. Their activities were nearly equal to that of epigallocatechin gallate. PMID- 12729011 TI - (-)-olivil and (+)-1-acetoxypinoresinol from the olive tree (Olea europaea Linne; Oleaceae) as feeding stimulants of the olive weevil (Dyscerus perforatus). AB - Guided by a feeding stimulant activity test on the olive weevil (Dyscerus perforatus), two compounds that showed potent feeding stimulant activity were isolated from the olive tree (Olea europaea). Based on their spectral data and a literature survey, they were identified as (-)-olivil (1) and (+ )-1 acetoxypinoresinol (2). The activities of these minor lignans were significantly higher for the female than for the male weevil. PMID- 12729012 TI - Effect of an oral administration of Lactobacillus casei strain Shirota on the natural killer activity of blood mononuclear cells in aged mice. AB - The natural killer (NK) activity of blood mononuclear cells and splenocytes in aged mice fed on a diet containing Lactobacillus casei strain Shirota (LcS group) was significantly (P < 0.01) higher than that in mice fed on a diet without LcS. In the LcS group, there was a significant positive correlation (r = 0.63, P < 0.01) between the NK activity of blood mononuclear cells and the NK activity of splenocytes. PMID- 12729013 TI - Inhibition of osteoporosis in rats fed with sugar cane wax. AB - Rats fed on a restricted, semi-purified diet containing a 50%-reduced level of carbohydrate and oil, but normal levels of protein, minerals and vitamins, exhibited osteoporosis. However, rats fed on this restricted diet, but containing sugar cane wax, did not exhibit this bone disease. Sugar cane wax, containing a long-chain carbohydrate with an OH radical, prevented the development of osteoporosis via a non-estrogenic mechanism. PMID- 12729014 TI - Release characteristics of flavor from spray-dried powder in boiling water and during rice cooking. AB - The release characteristics of flavor in boiling water and the flavor retention in the rice after cooking were investigated by using spray dried powder in encapsulated in or emulsified with d-limonene or ethyl n-hexanoate in cyclodextrin and maltodextrin, or in gum arabic and maltodextrin. The behavior of flavor release into the boiling water was well simulated by Avrami's equation. The retention of d-limonene and ethyl n-hexanoate in cooked rice was correlated in each case with the flavor amount of spray-dried powder added. PMID- 12729015 TI - Effect of dietary fiber on the lipid metabolism and immune function of aged Sprague-Dawley rats. AB - Eight-month-old Sprague-Dawley rats were fed on diets containing dietary fiber at the 5% level for 3 weeks to examine the effect on the lipid metabolism and immune function. Among cellulose, guar gum, partially hydrolyzed guar gum (PHGG), glucomannan and highly methoxylated pectin, guar gum induced a significant decrease in the food intake and weight gain, as well as a significant increase in the liver weight. In addition, the epidydimal adipose tissue weight of the rats fed on PHGG was significantly higher than that of the rats fed on cellulose. There was no significant effect on the serum lipid levels, but the serum IgG level of the rats fed on guar gum was significantly lower than that of the rats fed on cellulose. The IgA and IgG productivity in mesenteric lymph node (MLN) lymphocytes was significantly higher in the rats fed on guar gum, glucomannan and pectin than in those fed on cellulose, while the effect on Ig productivity in spleen lymphocytes was not as marked. In addition, only guar gum induced a significant increase of IgM productivity in MLN lymphocytes when compared to the cellulose group. These results suggest that enhancement of the immune function by dietary fiber is mainly expressed in the gut immune system. PMID- 12729017 TI - Cloning and overexpression of the 3-hydroxyisobutyrate dehydrogenase gene from pseudomonas putida E23. AB - The structural gene for NAD+-dependent 3-hydroxyisobutyrate dehydrogenase (EC 1.1.1.31) from Pseudomonas putida E23 was cloned in Escherichia coli cells to obtain a large amount of the enzyme and its nucleotides were sequenced to study its structural relationship with other proteins. The gene encoded a polypeptide containing 295 amino acid residues and was in a cluster with the gene for methylmalonate semialdehyde dehydrogenase. Transformed E. coli cells overproduced 3-hydroxyisobutyrate dehydrogenase, and the recombinant enzyme was purified to homogeneity with a high yield. Lysine and asparagine residues, which are important in catalysis of the 3-hydroxyacid dehydrogenase family, are conserved in this enzyme. PMID- 12729016 TI - Synthesis of hydroxymethylglutathione from glutathione and L-serine catalyzed by carboxypeptidase Y. AB - Hydroxymethylglutathione (gamma-L-glutamyl-L-cysteinyl-L-serine; hmGSH) occurs in many species belonging to the family Gramineae, but the biosynthetic pathway for hmGSH has not been identified. We found that carboxypeptidase Y (CPY), but not carboxypeptidase A, catalyzed hmGSH synthesis from glutathione and L-serine in vitro at acidic pH. CPY also catalyzed methylglutathione synthesis from glutathione and L-alanine. These findings suggested that a carboxypeptidase-like enzyme may be involved in hmGSH synthesis in vivo. PMID- 12729018 TI - Hydroxysulochrin, a tea pollen growth inhibitor from the fungus Aureobasidium sp. AB - A new plant growth regulator, hydroxysulochrin (1), together with sulochrin (2) was isolated from the culture filtrate of Aureobasidium sp. grown on a malt extract medium. The structures of 1 and 2 were established by spectroscopic methods. 1 and 2 inhibited tea pollen tube growth by 41% and 36% of the control value at a concentration of 100 mg/l, respectively. However, 1 and 2 showed no inhibitory effect on the growth of lettuce seedlings from 0.1 mg/l to 100 mg/l. PMID- 12729019 TI - 6-hydroxyflavonoids as alpha-glucosidase inhibitors from marjoram (Origanum majorana) leaves. AB - A methanol extract of marjoram leaves strongly inhibited rat intestinal alpha glucosidase. Five 6-hydroxyflavonoids, 6-hydroxyapigenin (scutellarein; IC50 for sucrose hydrolysis by rat intestinal alpha-glucosidase, 12 microM), 6 hydroxyapigenin-7-O-beta-D-glucopyranoside (> 500 microM), 6-hydroxyluteolin-7-O beta-D-glucopyranoside (300 microM), 6-hydroxyapigenin-7-O-(6-O-feruloyl)-beta-D glucopyranoside (>500 microM), and 6-hydroxyluteolin-7-O-(6-O-feruloyl)-beta-D glucopyranoside (> 500 microM), were isolated as active principles and related compounds. The two feruloylglucosides are novel compounds. PMID- 12729020 TI - Cooperation of Sly1/SM-family protein and sec18/NSF of Saccharomyces cerevisiae in disassembly of cis-SNARE membrane-protein complexes. AB - Assembly and disassembly of the SNARE membrane-protein complexes plays a key role in vesicular trafficking. The SM-family Slyl protein binds to the tSNARE Sed5 protein and stimulates its assembly into a trans-SNARE complex. Disassembly of the resulting cis-SNARE complex containing Sed5 was retarded in a temperature sensitive yeast mutant of Slyl protein with a defect in binding to Sed5. A temperature-sensitive mutation (sec18-1) of Sec18/NSF disassembly ATPase showed synthetic lethality with the sly1(ts) mutation. These results suggest that Slyl and Sec18 proteins work cooperatively and that the binding of Slyl to Sed5 stimulates the disassembly of the cis-SNARE complex by Sec18 ATPase. PMID- 12729021 TI - Evidence of isozymes for delta6 fatty acid desaturase in rat hepatocytes. AB - The expression of delta6 fatty acid desaturase, previously identified, was suppressed almost completely by hyper expression of the corresponding antisense gene in a transformant of the rat hepatic cell line BRL-3A. Conversion rates of [1-14C] linoleic acid, alpha-linolenic acid, and tetracosapentaenoic acid into the respective delta6 fatty acids were equivalent to those in control cells. This finding suggested that all of these reactions were catalyzed by at least two delta6 desaturase isozymes in rat hepatocytes. PMID- 12729022 TI - In vivo visualization of the distribution of a secretory protein in Aspergillus oryzae hyphae using the RntA-EGFP fusion protein. AB - A fusion gene encoding ribonuclease T1-EGFP (rntA-egfp) was constructed and expressed to use it as a tool for studies on the secretory pathway in Aspergillus oryzae. The successful secretion of the intact RntA-EGFP fusion protein was detected by fluorescence measurement and Western analysis. With use of the RntA EGFP system, we were able to see high fluorescence at hyphal tips and observe concentrated fluorescence at septa in basal cells during growth at optimal conditions. Cold or heat shock during growth caused the accumulation of EGFP fluorescence in vacuoles. PMID- 12729024 TI - Nursing--an ethic of caring. PMID- 12729023 TI - How is case management used with substance abuse cases? PMID- 12729025 TI - Safety is everyone's responsibility. PMID- 12729026 TI - Occupational health nurses with the certified safety professional credential. PMID- 12729028 TI - Integration of occupational health and safety: case study of a successful program. AB - 1. Occupational health nurses are key players in formulating a diagnosis, designing a plan of care, and implementing any interventions needed to promote safety. 2. While adhering to accepted confidentiality parameters, occupational health nurses share findings with appropriate individuals and teams when they are relevant to the health and safety of others. 3. No individual or department works alone to effect successful safety outcomes. Members of Occupational Health Services, Human Resources and Personnel, and the Safety Team depend on each other to improve work processes and create a safer work environment. PMID- 12729027 TI - Environmental, health, and safety management systems and auditing programs: part I--The evolution. AB - Early auditing began as an effort to avoid fines or other action from governmental agencies, without being based on accepted standards. However, for EHS auditing to be accepted as credible in the business world, established standards were necessary. As companies expanded globally, the need for international EHS standards grew, international standards for quality management and environmental program management have now been universally accepted (ISO, 2002). Occupational health nurses increasingly are becoming involved in efforts to help their employers or clients develop management systems to handle EHS issues--whether ISO 9000 (or the automotive equivalent, QS-9000), ISO 14000, or other models are used as the basis for the management system. Many nurses are actively involved in ISO certification efforts. As an extension of those efforts, occupational health nurses are increasingly involved in EHS audits, whether audits are conducted by third parties, by company employees, or as part of a self audit. The next column in this series will focus on strategies to improve the management of occupational health programs so the programs will stand up to rigorous EHS audits. PMID- 12729029 TI - The three Rs of fire safety, emergency action, and fire prevention planning: promoting safety at the worksite. AB - Fire safety is of paramount importance for everyone. In many workplaces, the occupational health nurse's scope of practice encompasses safety related activities. Included within this role is the responsibility for fire safety, emergency action, and fire prevention planning. The Three Rs of fire safety, emergency action, and fire prevention plans are rules, responsibilities, and resources. Myriad building and fire safety codes, regulations, and standards exist with which an employer must comply. An employer's responsibility for installing, testing, inspecting, and maintaining fire safety related equipment is extensive. Emergency action and fire prevention planning begins with conducting a detailed physical survey and preparing site maps. It includes making key policy decisions, writing procedures, and training employees in those procedures by practicing and executing site drills. The best resources available for emergency planning are the local fire department and the property insurer. Planning ahead means an efficient emergency response if disaster strikes. It saves lives, limits property damage, and preserves the environment. PMID- 12729030 TI - A primer for effective accident investigation: reducing the toll on resources. AB - Effective accident investigation is a combination of knowledge, skills, experience, and dedication to the mission of revealing the underlying factors of accidents. The purpose ultimately is to prevent similar accidents. Occupational health nurses are in a unique position by virtue of their education and skills to be an active participant in the process. PMID- 12729032 TI - Headache tips. PMID- 12729031 TI - Implementing a job hazard analysis program. AB - Analyzing the workplace for hazards and controlling the hazards identified to reduce risk is a proactive approach to workplace safety and the primary prevention of occupational illnesses and injuries. A JHA is a simple, time tested, and effective tool to assist in this analysis. Job hazard analysis methods may enhance and complement many current business initiatives. When programs can be integrated to contribute to leadership mandates, there are opportunities for increased support and resources. Involving other departments and disciplines with complementary programs also reduces duplication of effort. Prioritizing jobs to be analyzed based on risk assessment results in a manageable approach for beginning a program. A team process involving employees, supervisors, and key safety participants is most effective. Successful programs encourage each team member to observe or videotape key jobs, list the job steps, and involve the team in developing hazard reduction solutions and safe job procedures. When effectually used, JHAs have been proven to improve communication and participation, while enhancing worker safety. PMID- 12729033 TI - Clinical benefits of switching to an inhaled corticosteroid extrafine aerosol; a case series. AB - Hydrofluoroalkane beclomethasone dipropionate (HFA-BDP) extrafine aerosol is the first in a new generation of inhaled corticosteroid (ICS) formulations that have an improved deposition profile in comparison with conventional ICS preparations. This reformulation offers potential benefits to patients with asthma in terms of improved symptom control and reduced oropharyngeal adverse effects, such as dysphonia and candidiasis. This article presents four cases that illustrate the clinical benefits that can be obtained following a switch from conventional ICS preparations to treatment with HFA-BDP extrafine aerosol. The patients described were experiencing significant exacerbations of their asthma and increasing asthma symptoms and/or oropharyngeal adverse effects during treatment with conventional ICS preparations. On switching to HFA-BDP extrafine aerosol, the patients experienced an improvement in their asthma control and resolution of any oropharyngeal adverse effects. PMID- 12729034 TI - Changes in monocyte phagocyting activity after multi-agent chemotherapy in non small cell lung cancer. AB - Changes in monocyte functions have been described in several human malignancies. The monocyte/macrophage system is known to play a crucial role in the rejection of tumor cells and phagocytosis represents an important defense mechanism used by these cells. This paper reports the adherence power and phagocyting ability (latex beads) of circulating monocytes in 20 patients with unresectable non-small cell lung cancer (NSCLC), stage IIIB or stage IV, before and after multiagent chemotherapy (carboplatin + etoposide + ifosfamide or cisplatinum + etoposide). We demonstrated that both monocyte adherence and phagocytosis were not affected in lung cancer patients before chemotherapy in comparison with healthy controls. After chemotherapy, a statistically significant decrease in monocyte count on day 4 (p < 0.05) and in their phagocyting ability on day 4 and 15 (p < 0.001 and p < 0.05 respectively) was showed. In addition, a statistically reduced monocyte adherence was found on day 4 (p < 0.05). The described impairment was prolonged but reversible. These changes in monocyte functions after chemotherapy could be due to a direct effect of the chemotherapy on these cells or to functionally immature cells circulating after myelodepression. The in vitro assessment of monocyte functions may be useful to better clarify mechanisms by which anti neoplastic agents may act on immune functions and prevent adverse side effects. PMID- 12729035 TI - Potential implications for pathophysiology in a type 1 diabetic patient affected by Stargardt disease. AB - PURPOSE/METHOD: To present a 35-year-old woman affected by type 1 diabetes mellitus, Stargardt maculopathy and fundus flavimaculatus. To our knowledge, this association is unusual and not yet described in the ophthalmic literature. RESULTS/CONCLUSIONS: Visual acuity was reduced to < 20/200 in both eyes, color vision was absent and computerized perimetry showed an absolute central scotoma. Pattern visual evoked potentials and electroretinogram (ERG) (scotopic, photopic and flicker) were considerably reduced in amplitudes. Full-field ERG was within normal limits whereas oscillatory potentials were reduced in number and amplitude. Fluorescein angiography confirmed the diagnosis of Stargardt maculopathy and fundus flavimaculatus but no diabetic retinopathy was clinically evident. Potential interactions between the diabetic microangiopathy and the retinal degenerative disorder are hypothesized and discussed. PMID- 12729037 TI - The international year of freshwater 2003. PMID- 12729036 TI - Hyperhomocysteinemia in menopausal hypertension: an added risk factor and a dangerous association for organ damage. AB - Hyperhomocysteinemia is widely recognised as an emerging risk factor of endothelial dysfunction and vascular damage. In this study we wanted to verify if it, when associated to arterial hypertension--traditional risk factor--represents a higher added risk of organ damage during menopause, which is a condition connected to a higher incidence of cerebrovascular diseases. A survey of 30 postmenopausal women with similar characteristics (BMI, age, absence of relevant pathologies such as diabetes, metabolic disorders and absence of smoking) was selected (menopause had occurred from 12 to 16 months at the moment of observation). At the moment of the observation they had not gone through any continuous pharmacological therapy. They were subdivided into 3 groups: normotensive; hypertensive (with 2nd degree hypertension: mild to moderate) without organ damage; hypertensive with organ damage (TIA, ischaemic heart disease, etc.). The carotid IMT, measured with ultrasound method, was considered as an organ damage parameter. 43% of the patients had high levels of homocysteine (> 15 micromol/l), which are levels considered at risk in other surveys. The highest levels of homocysteine were recorded in hypertensive women with episodes of acute cerebrovascular damage (micromol/l = 24.3 +/- 8.9). In this group, a positive correlation (r = 0.7) was obtained between homocysteine levels and carotid IMT. The possible coexistence of hyperhomocysteinemia and arterial hypertension, even though without particularly high values for both of them, in menopause may represent a dangerous association responsible for a significant organ damage and, therefore, for acute cerebrovascular events. PMID- 12729038 TI - What is the lesson from the unprecedented event over Antarctica in 2002? AB - Varotsos (2002a,b), suggested that both the smaller-sized ozone hole over Antarctica and its splitting in two holes in September 2002 occurred due to an unprecedented major sudden stratospheric warming caused by very strong planetary waves propagated in the southern hemisphere. Subsequently, a NASA press release of December 6, 2002, also reported the prevalence of very strong planetary waves in Antarctica. The aim of this Letter is to further discuss the morphology of the Antarctic ozone hole, to detect the causes that allowed the Antarctic stratosphere to exhibit this exceptional warming and to examine what it denotes about its mechanisms. Concerning the morphology, among the principal findings is that the ozone hole split occurred not only in the stratosphere but extended in the lower altitudes (upper troposphere). As to the causes of the major sudden stratospheric warming of 2002, a comparison with the previous warmings in Antarctica since 1964 is made. The smaller-sized Antarctic ozone hole of 2002 is approximately equal to that of 1988 when a strong sudden stratospheric warming occurred. If only the destruction of ozone by chlorofluorocarbons resulted in the delayed sudden stratospheric warmings in Antarctica, then the early sudden stratospheric warmings of 1988 and 2002 would not have occurred, since chlorofluorocarbon loading of the stratosphere has remained relatively stable in recent years. Furthermore, it appears that the El Nino characteristics in 1988 and 2002 are not similar. PMID- 12729039 TI - Food irradiation: after 35 years, have we made progress. A government perspective. AB - The use of irradiation to improve the safety, protect the nutritional benefits, and preserve the quality of fresh and processed foods is a well established and proven technology. Over the past 35 years, the United States Government has invested in the science to confirm safety and in the technology to show application. The United States Department of Agriculture (USDA) and the Food and Drug Administration have approved sources of ionizing radiation for the treatment of foods, and their application to most meats, fruits, vegetables, and spices. Despite the value of this technology to the food industry and to the health and welfare of the public, only minimal application of this technology occurs. This underscores the importance of increasing the public's understanding of radiation risks relative to other hazards. Accordingly, in 1995, the Committee on Interagency Radiation Research and Policy Coordination of the Executive Office of the President made recommendations for the creation of a centralized National Radiation Information Center that would work closely with Federal departments and agencies in responding to public queries about radiation issues and Federal programs. This article updates a commentary published in 1996 (Young 1996). In the past six years, some progress has been made, including the establishment of a government operated Food Irradiation Information Center, and the completion of final rule making by USDA, thus permitting the safe treatment of meats and poultry. Despite these actions, little progress has been made on the public acceptance of this technology. The need for an informed public and for a better understanding of risks, i.e., risk communication, is noted. PMID- 12729040 TI - Effects of uncontrolled particulate matter release on precipitation under warfare conditions. AB - This study has shown theoretical, observed and experimental evidence of pollutants released, transported and deposited during the Kosovo conflict in 1999 and their effects on precipitation in Serbia. The greatest bombardment of the chemical industry, oil refineries and fuel storage in Serbia which occurred during April, resulted in releases of many hazardous, toxic and cancerogenic substances. The number of April's days with precipitation greater or equal to 0.1 mm in 1999 are compared to those in the period from 1961 to 1990 registered at thirty meteorological stations in Serbia and especially at the Belgrade Observatory station in the period from 1888 to 1995. The maximums of days with precipitation greater or equal to 0.1 mm were at the wider Belgrade area and at the central and southwestern parts of Serbia during April 1999. This is confirmed by using the Eta trajectory analysis. PMID- 12729041 TI - Determination of aliphatic and polycyclic aromatic hydrocarbons in atmospheric particulate samples of A Coruna City (Spain). AB - The hydrocarbon composition of atmospheric particulate matter from A Coruna city (Northwest Spain) has been studied. TSP (total suspended particulate) and PM10 (particulate matter with aerodynamic diameter < 10 microm) samples were taken over 7 and 4 months at two stations located in residential and industrial sectors of A Coruna city, respectively. The levels of hydrocarbons found in atmospheric particulate samples of A Coruna city are higher than the ones found in other cities over the world. Ratios between the analysed compounds and their sources were established. Both anthropogenic and biogenic origins were found. PMID- 12729042 TI - Effect of a heavy metal model mixture on biological parameters of rainbow trout Oncorhynchus mykiss. AB - The effects of a model mixture (HMMM) of seven heavy metals (Cu, Zn, Ni, Cr, Pb, Cd, Mn) on the rainbow trout Oncorhynchus mykiss at all stages of development (embryos, larvae, adults) were investigated based on the annual average concentrations of these metals in cooling waste waters discharging from Ignalina Nuclear Power Plant (Lithuania) into the Druksiai lake. According to mortality parameters, the most sensitive to HMMM were larvae, although no significant differences between the sensitivity of embryos and adult fish to HMMM were found. Maximal toxic effect of HMMM was observed during the hatching period. Long-term exposure to sublethal concentrations of HMMM affected embryo development, growth of larvae, their cardio-respiratory and behavioural responses, induced significant changes in morphological, morpho-physiological, physiological and haematological parameters of adult fish. Respiratory responses and growth parameters of fish were found to be the most sensitive to low concentrations of HMMM. Adult fish were capable of detecting and avoiding low, sublethal concentrations of HMMM. Heavy metals in a mixture at low concentrations were more toxic than single ones. According to the background of the damages induced by HMMM, after-effects in a fish organism, as well as in a whole population, can be predicted. PMID- 12729043 TI - Bioremediation of crude oil polluted soil by the white rot fungus, Pleurotus tuberregium (Fr.) Sing. AB - Bioremediation has become an attractive alternative to physicochemical methods of remediation of polluted sites. White rot fungi (WRF) are increasingly being investigated and used in bioremediation, because of their ability to degrade an extremely diverse range of very persistent or toxic environmental pollutants. The white rot fungus, Pleurotus tuberregium, was examined for its ability to ameliorate crude oil polluted soil. This was inferred from the ability of the polluted soil to support seed germination and seedling growth in Vigna unguiculata, at 0, 7 and 14 days post treatment. Results obtained from the present study showed that bioremediation of soil contaminated with crude oil was possible, especially when the fungus had been allowed to establish and fully colonize the substrate mixed with the soil. There were significant improvements in % germination, plant height and root elongation values of test plants, when seeds were planted 14 days post soil treatment. At 1 to 5% crude oil pollution, % germination values were comparable with the values in control plants in the 14 days treatment, and significantly higher than values obtained in the day 0 treatment. Also, at the highest level of crude oil pollution (15%), there was about 25% improvement in % germination value over the 0 day treatment. This trend of improvement in values was also observed for plant height, root elongation and biomass accumulation as well as decreased total hydrocarbon content. PMID- 12729044 TI - Photochemical degradation and mineralization of 4-chlorophenol. AB - INTENTION, GOAL, SCOPE, BACKGROUND: Since the intermediate products of some compounds can be more toxic and/or refractory than the original compund itself, the development of innovative oxidation technologies which are capable of transforming such compounds into harmless end products, is gaining more importance every day. Advanced oxidation processes are one of these technologies. However, it is necessary to optimize the reaction conditions for these technologies in order to be cost-effective. OBJECTIVE: The main objectives of this study were to see if complete mineralization of 4-chlorophenol with AOPs was possible using low pressure mercury vapour lamps, to make a comparison of different AOPs, to observe the effect of the existence of other ions on degradation efficiency and to optimize reaction conditions. METHODS: In this study, photochemical advanced oxidation processes (AOPs) utilizing the combinations of UV, UV/H2O2 and UV/H2O2/Fe2+ (photo-Fenton process) were investigated in labscale experiments for the degradation and mineralization of 4 chlorophenol. Evaluations were based on the reduction of 4-chlorophenol and total organic carbon. The major parameters investigated were the initial 4-chlorophenol concentration, pH, hydrogen peroxide and iron doses and the effect of the presence of radical scavengers. RESULTS AND DISCUSSION: It was observed that the 4-chlorophenol degradation efficiency decreased with increasing concentration and was independent of the initial solution pH in the UV process. 4-chlorophenol oxidation efficiency for an initial concentration of 100 mgl(-1) was around 89% after 300 min of irradiation in the UV process and no mineralization was achieved. The efficiency increased to > 99% with the UV/H2O2 process in 60 min of irradiation, although mineralization efficiency was still around 75% after 300 min of reaction time. Although the H2O2/4-CP molar ratio was kept constant, increasing initial 4-chlorophenol concentration decreased the treatment efficiency. It was observed that basic pHs were favourable in the UV/H2O2 process. The results showed that the photo-Fenton process was the most effective treatment process under acidic conditions. Complete disappearance of 100 mgl(-1) of 4-chlorophenol was achieved in 2.5 min and almost complete mineralization (96%) was also possible after only 45 min of irradiation. The efficiency was negatively affected from H2O2 in the UV/H2O2 process and Fe2+ in the photo-Fenton process over a certain concentration. The highest negative effect was observed with solutions containing PO4 triple ions. Required reaction times for complete disappearance of 100 mgl(-1) 4-chlorophenol increased from 2.5 min for an ion free solution to 30 min for solutions containing 100 mgl(-1) PO4 triple ion and from 45 min to more than 240 min for complete mineralization. The photodegradation of 4-chlorophenol was found to follow the first-order law. CONCLUSION: The results of this study showed that UV irradiation alone can degrade 4-CP, although at very slow rates, but cannot mineralize the compound. The addition of hydrogen peroxide to the system, the so-called UV/H2O2 process, significantly enhances the 4-CP degradation rate, but still requires relatively long reaction periods for complete mineralization. The photo-Fenton process, the combination of homogeneous systems of UV/H2O2/Fe2+ compounds, produces the highest photochemical elimination rate of 4-CP and complete mineralization is possible to achieve in quite shorter reaction periods when compared with the UV/H2O2 process. RECOMMENDATIONS AND OUTLOOK: It is more cost effective to use these processes for only purposes such as toxicity reduction, enhancement of biodegradability, decolorization and micropollutant removal. However the most important point is the optimization of the reaction conditions for the process of concern. In such a case, AOPs can be used in combination with a biological treatment systems as a pre- or post treatment unit providing the cheapest treatment option. The AOP applied, for instance, can be used for toxicity reduction and the biological unit for chemical oxygen demand (COD) removal. PMID- 12729045 TI - Dehalogenation potential of municipal waste incineration fly ash. II. Comparison of dehalogenation pathways of fly ash and model fly ash with thermodynamic calculations. AB - BACKGROUND, AIMS AND SCOPE: In the first part of this paper the main principles which control the dehalogenation of polychlorinated aromatic compounds on municipal waste incineration fly ash (MWI-FA) have been discussed and the model fly ash of similar dehalogenation activity has been proposed. Even if both systems show comparable dehalogenation properties, the main question concerning the postulated identical reaction mechanism in both cases is left unanswered. The other very important point is to what extent is this dechlorination mechanism thermodynamically controlled. The same problem is often discussed in the literature also for the de novo synthetic reactions. From the data it is clear that metallic copper plays a decisive role in the mechanism of the dehalogenation reaction. Although the results reported in the first part strongly support the idea that copper acts in this dechlorination as the reaction component, in contrast to its generally accepted catalytic behaviour, we believed that additional support for this conclusion can be obtained with the help of a thermodynamic interpretation of the mechanism of the reaction. RESULTS AND DISCUSSION: The pathways of hexachlorobenzene dechlorination on MWI-FA and model fly ash were studied in a closed system at 260-300 degrees C under nitrogen atmosphere. These pathways were the same for both systems, with the following prevailing sequences: hexachlorobenzene --> pentachlorobenzene --> 1,2,3,5 tetrachlorobenzene --> 1,3,5-trichlorobenzene --> 1,3-dichlorobenzene. Thermodynamic calculations were carried out by using the method of minimization total Gibbs energy of the whole system. In the calculations, the following reaction components were taken into account: all gaseous chlorinated benzenes, benzene, hydrogen chloride, a gaseous trimer Cu3Cl3, and also Cu2O and CuCl2 as solid components. The effect of the reaction temperature and the amount of copper and water vapour were considered as well. The effect of reaction temperature was determined from the data calculated for the 500 to 750 K temperature region. The effect of the initial composition was determined for the molar amounts of copper = 0.01-3 moles and water vapour = 0.2 to 3 moles per mole of chlorobenzene isomer CONCLUSIONS: The results of hexachlorobenzene dechlorination by MWI-FA and model fly ash under comparable reaction conditions allow us to conclude that both dechlorinations proceed via the same dechlorination pathways, which can be taken as an evidence of the identical dehalogenation mechanism for both systems. The relative percentual distribution of the dehalogenated products depends on the temperature, but not on the initial amount of water vapour or copper metal. On the other hand, the initial amount of copper substantially affects the conversion of the dehalogenation as well as the molar ratio of Cu3Cl3 to HCl in the equilibrium mixture. Comparison of the experimental with thermodynamic results supports the idea that dehalogenation reactions are thermodynamically controlled. RECOMMENDATIONS AND OUTLOOK: Thermodynamic analysis of the dehalogenation reactions may prove useful for a wide range of pollutants. The calculations concerning polychlorinated biphenyls and phenols are under study. PMID- 12729046 TI - Eutrophication of freshwater and coastal marine ecosystems: a global problem. AB - GOAL, SCOPE AND BACKGROUND: Humans now strongly influence almost every major aquatic ecosystem, and their activities have dramatically altered the fluxes of growth-limiting nutrients from the landscape to receiving waters. Unfortunately, these nutrient inputs have had profound negative effects upon the quality of surface waters worldwide. This review examines how eutrophication influences the biomass and species composition of algae in both freshwater and costal marine systems. MAIN FEATURES: An overview of recent advances in algae-related eutrophication research is presented. In freshwater systems, a summary is presented for lakes and reservoirs; streams and rivers; and wetlands. A brief summary is also presented for estuarine and coastal marine ecosystems. RESULTS: Eutrophication causes predictable increases in the biomass of algae in lakes and reservoirs; streams and rivers; wetlands; and coastal marine ecosystems. As in lakes, the response of suspended algae in large rivers to changes in nutrient loading may be hysteretic in some cases. The inhibitory effects of high concentrations of inorganic suspended solids on algal growth, which can be very evident in many reservoirs receiving high inputs of suspended soils, also potentially may occur in turbid rivers. Consistent and predictable eutrophication caused increases in cyanobacterial dominance of phytoplankton have been reported worldwide for natural lakes, and similar trends are reported here both for phytoplankton in turbid reservoirs, and for suspended algae in a large river CONCLUSIONS: A remarkable unity is evident in the global response of algal biomass to nitrogen and phosphorus availability in lakes and reservoirs; wetlands; streams and rivers; and coastal marine waters. The species composition of algal communities inhabiting the water column appears to respond similarly to nutrient loading, whether in lakes, reservoirs, or rivers. As is true of freshwater ecosystems, the recent literature suggests that coastal marine ecosystems will respond positively to nutrient loading control efforts. RECOMMENDATIONS AND OUTLOOK: Our understanding of freshwater eutrophication and its effects on algal-related water quality is strong and is advancing rapidly. However, our understanding of the effects of eutrophication on estuarine and coastal marine ecosystems is much more limited, and this gap represents an important future research need. Although coastal systems can be hydrologically complex, the biomass of marine phytoplankton nonetheless appears to respond sensitively and predictably to changes in the external supplies of nitrogen and phosphorus. These responses suggest that efforts to manage nutrient inputs to the seas will result in significant improvements in coastal zone water quality. Additional new efforts should be made to develop models that quantitatively link ecosystem-level responses to nutrient loading in both freshwater and marine systems. PMID- 12729047 TI - Acute renal failure complicating HELLP syndrome, SLE and anti-phospholipid syndrome: successful outcome using plasma exchange therapy. AB - Renal disease is very common in patients with systemic lupus erythematosis (SLE) and it may emerge during pregnancy or the post-partum period. Patients with anti phospholipid syndrome (APS)are also at risk of renal disease. We present a case of acute renal failure in the post-partum period in a patient with SLE and APS. This case illustrates the potential difficulties in reaching a diagnosis in such a patient. It also illustrates the complexities of management and the potential interactions between SLE, Haemolysis, elevated liver enzymes and low platelets syndrome, thrombotic thrombocytopenic purpura, haemolytic uraemic syndrome and APS. We also review the role of plasma exchange therapy in managing our patient. PMID- 12729049 TI - Use of a new fluorescence immunoassay to detect anti-dsDNA antibodies is more correlated with disease activity and complement than the ELISA method in SLE patients. AB - To determine whether the serum levels of anti-double strand DNA (anti-dsDNA) autoantibodies detected using a newly developed fluorescence immunoassay (FIA) in patients with systemic lupus erythematosus (SLE) correlated more with clinical parameters, such as SLE disease activity index (SLEDAI), complement and the occurrence of nephritis when compared with traditional enzyme-linked immunosorbent assay (ELISA), we prospectively collected 124 serum samples from 31 patients who had juvenile-onset SLE and were regularly monitored every 2 months at our outpatient clinic. At every visit, clinical manifestations and laboratory parameters were assessed and the SLEDAI was determined. Correlation analyses between the two different measurements of anti-dsDNA antibodies and SLEDAI, serum complement levels and the occurrence of nephritis were performed. The results showed that anti-dsDNA autoantibodies detected using both ELISA and FIA significantly correlated with SLEDAI, and significantly and inversely correlated with the serum levels of C3 and C4. FIA had significantly higher correlation with SLEDAI and C4 than did ELISA. The mean values of anti-dsDNA antibodies detected using FIA in patients with nephritis were significantly higher than in those without nephritis. In contrast, the values of anti-dsDNA antibodies detected using ELISA did not show significant differences between these two groups. We conclude that FIA had better correlation with SLEDAI, C4 and the occurrence of nephritis, and comparable correlations with C3 that were similar to the results found using ELISA. Thus, it is worthwhile developing the FIA method for clinical evaluation of disease activity in SLE patients. PMID- 12729048 TI - Association of the TAP2*Bky2 allele with presence of SS-A/Ro and other autoantibodies in Japanese patients with systemic lupus erythematosus. AB - We previously reported that a new allele of transporter associated with antigen processing (TAP) 2 gene, TAP2*Bky2 (Val577), was significantly increased in Japanese patients with Sjogren's syndrome (SS) and had a strong association with SS-A/Ro antibody production. In the present study, it was investigated whether the association of TAP2*Bky2 with SS-A/Ro antibody production was also found in Japanese patients with systemic lupus erythematosus (SLE). Polymorphisms of the TAP1 and TAP2 genes were determined in 114 Japanese SLE patients by the polymerase chain reaction-single-stranded conformation polymorphism (PCR-SSCP) method. The allele frequencies of the TAP1 and TAP2 genes in SLE patients were not significantly different from those in controls, although the allele frequency of TAP2*Bky2 was slightly higher in SLE patients than in healthy control subjects (9.2% vs 5.5%, P = 0.126). The allele frequency of TAP2*Bky2 was significantly higher in SLE patients with oral ulcers than in those without. It was noteworthy that TAP2*Bky2 was significantly associated with the appearance of not only SS A/Ro antibody but also SS-B/La, nRNP, and Sm antibodies in the patients. The association of TAP2*Bky2 was found with the antibody production to both 60 and 52kDa SS-A/Ro antigens. As TAP2*Bky2 had a strong linkage disequilibrium with DRB1*08032, TAP2*Bky2 or its haplotype with DRB1*08032 may be involved in SS-A/Ro antibody production not only in SS but also SLE patients, indicating that TAP2*Bky2 may be a susceptible gene not only to the disease of SS but also to the SS-A/Ro autoantibody production. PMID- 12729050 TI - Systemic lupus erythematosus in three ethnic groups: XV. Prevalence and correlates of fibromyalgia. AB - The purpose of this study was to determine the prevalence and correlates of fibromyalgia (FM) in a prospective, multiethnic systemic lupus (SLE) cohort. A total of 266 SLE patients with disease duration of < or = 5 years at study entry were evaluated longitudinally for the presence of FM (per ACR criteria). Sociodemographic factors, behavioral/psychological variables, clinical features, serologic factors (autoantibodies), and self-reported functioning (MOS SF-36) were ascertained in all patients. Subjects were evaluated at study entry and annually thereafter. The prevalence of FM was then calculated, as was the prevalence of FM-like manifestations (widespread pain with at least 6, but fewer than 11/18 tender points). Variables were evaluated for association with FM or FM like manifestations by univariate and stepwise logistic regression analyses. FM was present in 14 patients (5%; 9/92 Caucasians (C), 4/109 African Americans (AA), 1/65 Hispanics (H)) and FM/FM-like manifestations in 35 (13%; 16 C, 9 AA, 10 H). There was no difference noted between those with and without FM with respect to gender, education level, income below poverty level, disease activity or damage. By stepwise logistic regression analyses, the strongest association with both FM and FM/FM-like manifestations was a self-reported history of anxiety or affective disorder (P = 0.0237, OR = 4.6 and P = 0.0068, OR = 3.4, respectively). Caucasian ethnicity was strongly associated with FM (P = 0.0066, OR = 7.5) and African American ethnicity was negatively associated with FM/FM like (P = 0.0204, OR = 0.3). Poorer self-reported physical functioning was associated with FM/FM-like (P = 0.0443, OR = 0.96). FM and FM-like manifestations correlate best with the presence of Caucasian ethnicity, concomitant anxiety or affective disorder, and to a lesser extent with poorer self-reported physical functioning. African American ethnicity is negatively associated with the combination of FM and FM-like manifestations. Clinical measures of disease activity, disease damage, specific organ dysfunction, sociodemographic factors and serologic features are not correlated with FM in this early SLE cohort. PMID- 12729051 TI - Validation of a Systemic Lupus Activity Questionnaire (SLAQ) for population studies. AB - The goal of this work was to develop an economical way of tracking disease activity for large groups of systemic lupus erythematosus (SLE) patients in clinical studies. A Systemic Lupus Activity Questionnaire (SLAQ) was developed to screen for possible disease activity using items from the Systemic Lupus Activity Measure (SLAM) and tested for its measurement properties. The SLAQ was completed by 93 SLE patients just prior to a scheduled visit. At the visit, a rheumatologist, blinded to SLAQ results, examined the subject and completed a SLAM. Associations among SLAQ, and SLAM (omitting laboratory items) and between individual items from each instrument were assessed with Pearson correlations. Correlations between pairs of instruments were compared using Student's t-tests. The mean score across all 24 SLAQ items was 11.5 (range 0-33); mean SLAM without labs was 3.0 (range 0-13). The SLAQ had a moderately high correlation with SLAM nolab (r = 0.62, P < 0.0001). Correlations between patient-clinician matched pairs of items ranged from r = 0.06 to 0.71. Positive predictive values for the SLAQ ranged from 56 to 89% for detecting clinically significant disease activity. In studies of SLE, symptoms suggesting disease can be screened by self-report using the SLAQ and then verified by further evaluation. PMID- 12729052 TI - Predictors of poor renal outcome in patients with lupus nephritis treated with combined pulses of cyclophosphamide and methylprednisolone. AB - Lupus nephritis remains a major cause of morbidity and mortality in patients with systemic lupus erythematosus. Although the renal prognosis has improved, the optimal therapeutic regime has not been definitively established, and significant challenges remain in the management of disease progression and recurrent renal relapse. We performed a prospective study to evaluate the outcome of 38 patients with severe lupus nephritis treated with standard cyclophosphamide and methylprednisolone pulse therapy, and to determine the variables associated with poor outcome. Five patients developed end-stage renal disease (ESRD) (13%), 10 (26%) developed persistent proteinuria (> 1 g/24h) and 15 (39%) suffered at least one relapse after 8 years of follow-up. A high chronicity index, interstitial fibrosis (P = 0.04), persistent hypertension (P < 0.0001) and hypocomplementaemia (P = 0.002) after treatment were the major variables associated with ESRD. Tubular atrophy (P = 0.01), persistent hypertension (P = 0.0001) and hypocomplementaemia after treatment (P = 0.0281) were associated with persistent proteinuria. Persistence of anti-dsDNA antibodies and hypocomplementaemia after treatment (P = 0.0118) were associated with renal relapse. Our data suggest that the group of patients with persistence of hypocomplementaemia and raised anti dsDNA antibodies titres are at high risk of renal relapse and may be candidates for continuation of immunosuppressive treatment. Patients with persistent proteinuria alone or a high chronicity index are less likely to respond to immunosuppression, and strict control of the hypertension may be the best approach. PMID- 12729053 TI - Quinacrine added to ongoing therapeutic regimens attenuates anticardiolipin antibody production in SLE. AB - The benefit of combining quinacrine (Qn) with hydroxychloroquine (HCQ) in the treatment of systemic lupus erythematosus (SLE) was previously re-evaluated by us. In our current study we observed that, in 11 active SLE patients (SLEDAI score 5-12), the addition of Qn (100 mg/day) to their existing ongoing therapeutic regimens resulted in a significant attenuation of their previously persistent anticardiolipin antibody (aCL) response. This was in comparison with a matched non-Qn treated control group composed of 14 randomly chosen aCL-positive SLE patients with a similar SLEDAI score 6-10. Prior to Qn treatment the therapeutic regimens of 12 months' duration, included in all cases HCQ (400 mg/day), in many cases prednisone (P, 10-20 mg/day) and in some additional cases immunosuppressive drugs. SLEDAI scores and aCL levels were monitored during the entire follow-up period which totaled 24 months in the study group and 15-18 months in the controls. Along with the beneficial effect of the added Qn on SLEDAI scores, aCL disappearance was documented in eight of 11 patients and remained negative during 8-12 months of follow-up (P = 0.004), compared with such a change in only three of 14 non-Qn treated aCL-positive patients (P = 0.18). We conclude that the added Qn treatment to former established therapeutic protocols may eliminate aCL response in SLE patients. Whether this agent's effect is permanent needs further elucidation. PMID- 12729054 TI - Evaluation of clinical and laboratory features of antiphospholipid syndrome: a retrospective study of 637 patients. AB - We retrospectively analysed the data of 1519 antiphospholipid antibody (APLA) positive patients between 1986 and 1999. Among them 637 were considered to have antiphospholipid syndrome (APS) based on the 1999 preliminary classification criteria, while 704 patients had no clinical signs of the syndrome. Our aim was to compare the autoantibody profile and clinical characteristics of primary and secondary APS, moreover to evaluate the associations between different APLA and specific symptoms attributable to APS. In our results, the APLA profiles for primary and SLE-associated secondary APS were similar. Among the evaluated clinical symptoms, cerebrovascular thrombosis was found to be more frequent in the SLE-associated, than in the primary APS group (P = 0.04). We identified important differences in the clinical profile of patient populations with various types of APLA. Venous thrombosis occurred more frequently in subjects withlupus anticoagulant (LA), than in those with IgG or IgM type ACLA (P < 0.0001), while coronary, carotid and peripheral artery thrombosis occurred more often in subjects with IgG or IgM ACLA (P < 0.0001). These findings may support the role of antibodies to cardiolipin or its cofactor, beta2glycoprotein I (beta2-GPI) in the initiation and progression of atherosclerosis. Cerebrovascular thrombosis was detected in larger proportion of LA or IgG ACLA-positive patients compared with to IgM ACLA-positive subjects, while the occurrence of foetal loss was similar in all three groups. PMID- 12729056 TI - Mycobacterium avium complex-associated hemophagocytic syndrome in systemic lupus erythematosus patient: report of one case. AB - Hemophagocytic syndrome (HPS) in systemic lupus erythematosus(SLE) patients has not commonly been reported. In this case study, we report the first case of Mycobacterium avium complex (MAC)-associated hemophagocytic syndrome in a patient with systemic lupus erythematosus (SLE). This SLE patient, a 15-year-old girl, had been on a high dose of prednisolone (> 0.5mg/kg/day) for more than 3 years. She presented with a spiking fever, hepatosplenomegaly, pancytopenia, hyperferritinemia and adult respiratory distress syndrome. Bone marrow examination revealed hemophagocytosis as well as non-caseating granulomatosis. There was no indication of SLE fare-up. She responded poorly to initial treatment with methyl-prednisolone, intravenous immumoglobulin, etoposide, and drugs for Mycobacterium tuberculosis including rifampin, ethambutol, isoniazid and pyramide. However, gastric lavage culture revealed MAC. Following treatment with clarithromycin, ciprofloxacin and amikacin, her condition gradually improved and she was discharged 3 months after admission. In SLE patients with pancytopenia and hyperferritinemia, MAC-associated HPS should be considered in the differential diagnosis. PMID- 12729055 TI - Mitral valve prolapse in systemic lupus erythematosus patients: clinical and immunological aspects. AB - Mitral valve prolapse (MVP) has been reported to be associated with systemic lupus erythematosus (SLE). The aim of the present study was to determine the prevalence of MVP in SLE patients, assess its clinical significance and examine the possible association of this entity with other autoimmune indices. Eighty seven consecutive SLE patients attending the rheumatology clinic and 73 normal control subjects were examined by M-mode, two-dimensional color-Doppler echocardiography. Serum samples were examined for various organ and non-organ specific autoantibodies. MVP was detected in 19/87 patients with SLE and in four of the healthy controls(P = 0.0057). SLE patients with MVP were younger (33.6 +/- 12.4 years) than those without MVP (41. +/- 12.9, P = 0.04) and with shorter duration of the disease (P = 0.03). We found a statistically higher prevalence of anticardiolipin antibodies (aCL) in SLE patients with prolapse (11/19) compared with SLE patients without prolapse (15/68, P = 0.04). This association was independent of age. The aCL-lgG levels were significantly higher in SLE patients with MVP (32.37 +/- 43.26) compared with SLE patients without MVP (22.24 +/- 29.95, P = 0.04). Thyroid autoantibodies tended to be more common in S LE patients with MVP. Th e prevalence of MVP is increased in SLE patients. The presence of aCL and of organ-specific autoantibodies in SLE patients with MVP might indicate the autoimmune origin of MVP. The possibility that SLE patients with MVP may be predisposed to further autoimmune diseases should be considered. PMID- 12729057 TI - Ruptured renal microaneurysms complicated with a retroperitoneal abscess for a patient with systemic lupus erythematosus. AB - Renal artery aneurysm is extremely rare among patients with systemic lupus erythematosus.(SLE). Herein, we report on a 22-year-old male lupus patient who presented with acute abdominal pain, anemia and subsequent hypertension. Abdominal computed tomography revealed a peri-renal hematoma over the right kidney. A renal angiography revealed bilateral renal microaneurysms. The patient subsequently developed a right-side retroperitoneal abscess 4 weeks after hematoma formation and received an emergent laparotomy with drainage. Subsequent culture ofthe abscess-derived fluid revealed the presence of Proteus mirabilis and Escherichia coli. Following appropriate antipyretic and immunosuppressive drugs therapy, the patient recovered successfully. To the best of our knowledge, this is the first report of SLE associated with a retro-peritoneal abscess probably secondary to a ruptured renal microaneurysm. PMID- 12729058 TI - Systemic lupus erythematosus-associated pulmonary hypertension: good outcome following sildenafil therapy. AB - A 46-year-old woman with systemic lupus erythematosus (SLE) and concomitant severe pulmonary hypertension (PH) is described. Other secondary causes of PH including thromboembolism, phospholipid syndrome, valvular disease and interstitial pulmonary involvement were ruled out. Owing to her lack of clinical response to conventional therapy, sildenafil was begun at increasing doses up to 400 mg daily. Both clinical and hemodynamic improvement ensued. This appears to be the first clinical report of the use of sildenafil in SLE followed by resolution of severe PH. PMID- 12729059 TI - Chronic monoarthritis of the knee in systemic lupus erythematosus. AB - We describe a middle-aged lady with systemic lupus erythematosus who presented with chronic left knee monoarthritis without constitutional symptoms. The histology of synovial tissue taken at arthroscopy showed acute inflammation and Erysipelothrix rhusiopathiae, identified with some difficulty, was isolated from the enrichment broth only. Blood cultures were negative. Her history revealed significant exposure to pond fish. She responded well to intravenous penicillin and remains well 12 months later. PMID- 12729060 TI - Systemic lupus erythematosus in the far north of Queensland. AB - An assessment of prevalence for systemic lupus erythematosus (SLE) has been attempted for the population of far north Queensland in Australia. This huge area has a majority Caucasian population living in a tropical environment. Roughly 10% of the population comprises people of Australian Aboriginal or Torres Strait Islander descent. The prevalence of disease was high (45.3 per 100000) overall and particularly so in the indigenous population (92.8 per 100000) where the disease appears to be more severe. The pattern of organ involvement and laboratory anomalies did not vary between the populations examined. Disease duration however was different, being longer by comparison in the Caucasian population due to many premature deaths in the indigenous groups. The reasons for such high prevalence figures and some problems encountered in practice are discussed. PMID- 12729061 TI - Disease progression in the systemic lupus erythematosus model mouse (NZB/NZW F1) is inhibited by single shot estrogen treatment in the neonate. PMID- 12729062 TI - Anti-Ro and anti-nRNP response in unaffected family members of SLE patients. PMID- 12729063 TI - Starling, his contemporaries and the Nobel Prize. One hundred years with hormones. PMID- 12729064 TI - Biochemical values in persons older than 82 years of age: report from a population-based study of twins. AB - According to international standards, reference values for biochemical tests should be obtained in disease-free subjects and for therapies that may influence measurement values. For elderly persons, especially the very old, such a requirement is difficult to meet, since few of these individuals are free of ongoing or former diseases. The present study of a population-based Swedish twin sample comprises 535 individuals (186 males, 349 females) who were at least 82 years of age at blood sampling. Survival over a 6-year period is used as a reference of overall health. Baseline test values for subjects surviving 6 years are compared with results for persons who died in the follow-up period. Results, including mean and median values, standard deviations and ranges, were given for both genders in the total sample. Cases outside the 5th and 95th percentiles were then excluded, as were those who died during the follow-up period. In a third step, cases with disorders and therapies known to influence measurement values were also excluded. The risk of dying during the observation period was calculated across measurements from the lowest and highest 10th and 20th percentiles. The specific impact of proximity to death on the biochemical values, in addition to age effects, was analysed by comparing co-twins discordant for survival over 7 years. Increased mortality was indicated for subjects of both genders with high serum levels of urea, urate, gamma-GT, free thyroxin and plasma homocysteine. In females, low serum values for albumin and total cholesterol were associated with increased mortality. A lower cholesterol level was found 4 years before death, while a lower HDL-cholesterol level was associated with an increased risk of death across the entire 7-year period. PMID- 12729065 TI - Biological variation of soluble CD163. AB - A soluble plasma form of CD163 (sCD163) was recently identified. The protein has anti-inflammatory effects in vitro and is elevated in patients with myelo monocytic leukaemia and infection. For rational use and evaluation of this potential new quantity it is important to have knowledge of its biological variability and to use a methodology that has a sufficiently analytical quality. The day-to-day and diurnal biological variabilities of sCD163 were studied in 12 healthy people using a sandwich ELISA. The within-run-, between run- and total analytical coefficients of variation were estimated to 3.6%, 4.8% and 6.0%, respectively. The day-to-day within-subject biological variation was estimated to 9.0%, and the between-subject biological variation to 35.9%. A diurnal variation in sCD163 concentrations with 14% lower values in the night (supine position) was observed. The ratio between total analytical variation and within-subject biological variation was 0.67. The index of individuality, calculated as the ratio between within-subject biological variation and between-subject biological variation, was low and similar to complement factors and immunoglobulins. A low index of individuality is important in a monitoring situation where small changes from the set point of the individual can be detected in serial measurements. For this purpose, the critical difference for a series of results in the same patient (significant at p < 0.05) was calculated to 30% for samples taken on different days and measured in separate runs. PMID- 12729066 TI - Androgen levels in pregnant women decrease with increasing maternal age. AB - BACKGROUND: To investigate a possible effect of age on maternal androgen levels in uncomplicated pregnancies. METHODS: A study of 134 parous women with uncomplicated pregnancies was carried out at three university hospitals in Norway and Sweden. Maternal levels of androstenedione, dehydroepiandrosterone sulphate, testosterone and the free testosterone index were measured during weeks 17 and 33 of pregnancy. RESULTS: Maternal levels of androstenedione and testosterone had a negative association with maternal age in weeks 17 and 33 of pregnancy, while dehydroepiandrosterone sulphate and the free testosterone index were associated negatively in week 33 only. Adjustment for maternal parity, pre-pregnancy body mass index, smoking and fetal gender did not affect the results. CONCLUSIONS: Maternal androgen levels decrease with increasing maternal age. The cause and possible implication of this finding remain unknown. PMID- 12729067 TI - Studies of cytokine levels in bronchoalveolar fluid lavage from patients with interstitial lung diseases. AB - Cytokine levels in bronchoalveolar lavage fluid from patients with eosinophilic pneumonia (n = 7), allergic alveolitis (n = 11), (cryptogenic) fibrosing alveolitis (n = 8), sarcoidosis (n = 10) were determined, as well as levels in control samples from healthy non-smoking volunteers (n = 11). Fibronectin levels were increased in all the patient categories, the highest absolute levels of fibronectin (100-fold increase) being found in eosinophilic pneumonia and allergic alveolitis. TGF-beta (transforming growth factor-beta) was significantly elevated in allergic alveolitis only. There was a significant difference between allergic alveolitis on the one hand and both sarcoidosis and fibrosing alveolitis on the other. Tumour necrosis factor-alpha (TNF-alpha) was significantly increased in eosinophilic pneumonia and allergic alveolitis; allergic alveolitis and fibrosing alveolitis differed significantly in this respect. Platelet-derived growth factor-BB (PDGF-BB) levels were significantly elevated in allergic alveolitis and fibrosing alveolitis. It was found that the level of PDGF-BB was significantly decreased in the case of sarcoidosis, with no overlapping with allergic alveolitis or fibrosing alveolitis. Interferon-gamma (IFN-gamma) was decreased in all patient categories. A significant difference in extent of the decrease was found between allergic alveolitis and sarcoidosis. The interstitial lung diseases thus differed in the pattern of cytokines expressed, indicating that these cytokines could well be a part of the pathogenic process, and also that the measurement of cytokine levels could be diagnostically useful. PMID- 12729068 TI - Pyridinoline cross-links as markers for primary and secondary bone tumors. AB - BACKGROUND: Determination of hydroxylysylpyridinoline (HP) and lysylpyridinoline (LP) in urine is a promising method to determine bone resorption. This method is independent of gender, diet and kidney function (creatinine clearance > 25 mL/min). METHODS: The diagnostic efficacy of HP and LP was assessed in the urine of adult patients suffering from primary malignant bone tumors (n = 24), bone metastases (n = 38) and soft tissue sarcoma with additional osseous involvement (n = 13). The values were compared with those obtained from 543 healthy controls (aged 15 to 65 years). RESULTS: Results clearly exhibited a highly significant increase in HP values (57.75 nmol/mmol creatinine) in adult tumor patients (aged 15 to 65 years) in all three subgroups in comparison with the control group values (22.23 nmol/mmol creatinine) (p = 0.001). Although the LP fraction is more specific for bone than HP, the values of LP from all subgroups of the adult tumor patients were less distinctly but still significantly increased (p = 0.008). Regarding the HP:LP ratio, tumor patients exhibited a markedly increased average molar HP:LP ratio (12.0:1) in comparison to controls (6.6:1). CONCLUSION: Determination of HP and LP in urine appears to offer clinical utility for the detection of primary and secondary bone neoplasms. Prospective, longitudinal studies will be necessary to evaluate whether the HP:LP ratio is indicative of a relapse of the tumor and consecutively may be adopted in the follow-up of patients with neoplasms. PMID- 12729069 TI - Lipoprotein lipase during heparin infusion: lower activity in hemodialysis patients. AB - BACKGROUND: [corrected] Patients on hemodialysis often have a moderate hypertriglyceridemia in combination with low HDL cholesterol. A contributing factor may be a derangement of the lipoprotein lipase (LPL) system. During dialysis, with heparin as anticoagulant, the enzyme is released into the circulating blood. METHODS: We have followed LPL activity and triglycerides during ordinary heparin administration in nine hemodialysis patients and controls matched for age and gender. Blood samples were drawn before heparin administration and at 15, 30, 60, 120, 180 and 240 min. RESULTS: LPL activity peaked at 15 or 30 min and then decreased to a plateau that was only 20%, of the peak. The activity was reduced in the patients by about 50% during the peak, and about 20% during the following plateau. During the peak of lipase activity the triglycerides decreased in both groups, but the change was less pronounced in patients, as was expected from the lower circulating lipase activity. During the plateau phase with low lipase activity, the triglycerides increased towards baseline values. CONCLUSIONS: During hemodialysis with heparin, there is a peak in LPL activity as well as a reduction in triglycerides during the first hour. Thereafter LPL activity decreases towards a plateau, while triglycerides increase towards baseline. The peak activity of LPL in the patients was only half that in controls, while the plateau was comparable. The data indicate that during and following each dialysis there is a period when LPL activity becomes depleted to a level that is limiting for normal lipoprotein metabolism. PMID- 12729070 TI - Analytical performance of time-resolved fluorometry-based Innotrac Aio! cardiac marker immunoassays. AB - The results of an evaluation of the Innotrac Aio! cardiac markers are presented. This system is based on dry-chemistry, time-resolved fluorometry. All assay specific reagents are dry-coated into assay-specific cups, and only the generic assay buffer is required. The levels of precision attained with pooled serum samples and control materials were acceptable for cTnI and CK-MB. Myoglobin assay showed higher CV, 5.6-9.5%. The linearity studies were performed in concentration ranges of 0.1-76 microg/L for cTnI, 0.7-450 microg/L for CK-MB and 0.6-1500 microg/L for myoglobin. The markers were found to be linear within the ranges tested. The correlation coefficient between the Aio! and AxSYM cTnI assays was 0.960, and the slope was 0.07. The correlation coefficients between the Aio! and AxSYM CK-MB and myoglobin assays were 0.995 and 0.971, respectively. They involved some differences in the measured concentrations (Aio! CK-MB was about 9% higher than AxSYM CK-MB, and Aio! myoglobin was 19% higher than AxSYM). Comparative studies with all the markers, using EDTA whole blood and lithium heparin plasma specimens and lithium heparin whole blood and plasma, yielded the following results: the slopes were close to 1.0 for all correlations, with the exception of that between CK-MB EDTA whole blood and lithium heparin (0.83). High correlation coefficients were obtained (> or = 0.97). The carryover results for all the cardiac markers were good, 0.0%, 0.0%, and 0.3% for cTnI, CK-MB, and myoglobin, respectively. The analytical detection limits were 0.01 microg/L for cTnI, 0.8 microg/L for CK-MB and 0.5 microg/L for myoglobin. The stability of the analytes in the lithium heparin samples at room temperature was also studied and was found to be decreased by from 10% (myoglobin and CK-MB) to 17% (cTnI) in 8 h. Innotrac Aio! provides a rapid and easy quantitative measurement of cardiac TnI, CK-MB, and myoglobin within < 18 min. This system is therefore suitable for use in emergency departments, coronary care units or central laboratory settings. PMID- 12729072 TI - Plasma lipid and lipoprotein responses during exercise. AB - The purpose of this study was to examine the effect of prolonged exercise on plasma lipid and lipoprotein concentrations and to identify caloric time-points where changes occurred. Eleven active male subjects ran on a treadmill at 70% of maximal fitness (VO2max) and expended 6278.7 kilojoules (Kj) energy (1500 kcal). Blood samples were obtained at the 4185.8 Kj (1000 kcal) timepoint during exercise and at each additional 418.6 Kj (100 kcal) expenditure until 6278.7 Kj was expended. After correcting for plasma volume changes, decreases in low density lipoprotein cholesterol (LDL-C) were observed during exercise at time points corresponding to 4604.4 and 5441.5 Kj (1100 and 1300 kcal) of energy expenditure, and immediately after exercise. Total cholesterol concentrations decreased significantly at exercise kilojoule expenditures of 4604.4, 5441.5 and 5860.1 (1100, 1300 and 1400 kcal). There were also exercise induced increases in high-density lipoprotein cholesterol (HDL-C) and HDL2-C concentrations immediately after exercise. Although acute lipid and lipoprotein changes are typically reported in the days following exercise, the current data indicate that some lipoprotein concentrations change during acute exercise. Our data suggest that a threshold of exercise may be necessary to change lipoproteins during exercise. Future work should identify potential mechanisms (lipoprotein lipase, cholesterol ester transport protein, LDL uptake) that alter lipoprotein concentrations during prolonged exercise. PMID- 12729071 TI - Correlation of transthoracic Doppler echocardiography and magnetic resonance imaging in measuring left anterior descending artery flow velocity and time course of dipyridamole-induced coronary flow increase. AB - There is an increasing need for new, functional and more quantitative parameters to assess coronary arterial function, for the purposes of evidence-based medicine. Coronary arterial function has been widely studied using pharmacological stimulation induced by dipyridamole or adenosine. Coronary flow reserve (CFR), defined as the ratio of pharmacologically induced hyperemic flow divided by basal flow, has been found to be an important functional index in both the clinical and subclinical stages of cardiovascular diseases. Ten healthy male volunteers were studied to compare transthoracic Doppler echocardiography (TTE) and MRI for measuring left anterior descending coronary artery (LAD) flow velocity and CFR. Additionally, the time-response curve of dipyridamole infusion was studied in five healthy males using TTE. Assessment of blood flow velocity, measured as MDV, PDV and VTI indicated Pearson's correlation coefficients of 0.88, 0.85 and 0.70, respectively, between flow velocity measurements performed using TTE and MRI. The results indicate that, despite minor differences in LAD diastolic velocities measured by TTE and MRI, the correlation of the LAD diastolic velocities measured using both methods are good and both methods are feasible for measuring CFR. Moreover, TTE has the unique capability of continuous measurement of LAD flow velocity, which allowed assessment of the time-response curve for dipyridamole-induced increase in LAD flow velocity in this study. This study indicates that the TTE method may be used in sequential or on-line monitoring of LAD blood flow velocity and therefore can be applied to evaluate the time- or dose-response effects of infused drugs in the coronary circulation of humans. PMID- 12729073 TI - Determination of melatonin in human plasma with solid-phase extraction, high performance liquid chromatography and fluorescence detection. AB - A new bioanalytical method for the determination of melatonin in plasma with high performance liquid chromatography (HPLC) and fluorescence detection preceded by solid-phase extraction has been developed and validated. Melatonin was extracted from 3 mL plasma using a Waters Oasis HLB solid-phase extraction cartridge and the elute was evaporated to dryness and dissolved in 200 microl mobile phase; acetonitrile-phosphate buffer, 0.01 M pH 7.2 (25:75, v/v). 125 microL was injected into the HPLC system and separation was carried out on a Waters SymmetryShield RP18 column 5 microm (250 x 4.6 mm). Excitation and emission wavelengths were set to 285 nm and 345 nm, respectively. The HPLC system was able to separate melatonin and internal standard (5-fluorotryptamine) from other endogenous indole compounds such as serotonin and tryptophan. Determination down to 0.10 nmol/L was possible, with an intra-assay precision of about 13%. Melatonin was stable in plasma for at least 30 days at about 23 degrees C. PMID- 12729074 TI - Update: outbreak of severe acute respiratory syndrome--worldwide, 2003. AB - CDC and the World Health Qrganization (WHO) are continuing to investigate the multicountry outbreak of unexplained atypical pneumonia referred to as severe acute respiratory syndrome (SARS). Pending development of confirmatory laboratory testing capacity, CDC's interim suspected SARS case definition is based on clinical criteria and epidemiologic linkage to other SARS cases or areas with community transmission of SARS. This case definition will be updated periodically as new information becomes available. Epidemiologic and laboratory investigations of SARS are ongoing. As of April 2, 2003, a total of 2,223 suspected and/or probable SARS cases have been reported to WHO from 16 countries, including the United States. The reported SARS cases include 78 deaths (case-fatality proportion: 3.5%). This report summarizes SARS cases among U.S. residents and surveillance and prevention activities in the United States. PMID- 12729076 TI - Nonfatal fall-related traumatic brain injury among older adults--California, 1996 1999. AB - In the United States, falls are the second leading cause of traumatic brain injury (TBI) hospitalizations overall and the leading cause of TBI hospitalizations among persons aged > or = 65 years. In 1995, TBIs resulted in an estimated $56 billion in direct and indirect costs in the United States. In California, during 1999, a total of 61,475 hospitalizations from falls were reported among persons aged > 65 years. Risk factors for falling among older persons included arthritis; impairments in balance, gait, vision, and muscle strength; and the use of four or more prescription medications. As part of CDC's program of state-based TBI surveillance, California hospital discharge data were collected and analyzed to describe fall-related TBIs. This report summarizes the results of that analysis, which support previous findings that persons aged > or = 65 years are at risk for hospitalization for a fall and that same-level falls are far more common among persons aged > or = 65 years than falls from a higher level (e.g., a ladder, chair, or stair). Defining the circumstances of fall injuries and recognizing the type of fall leading to TBI hospitalizations among older persons can help health-care providers conduct risk assessment and management of falls in this population. PMID- 12729075 TI - Hepatitis C virus transmission from an antibody-negative organ and tissue donor- United States, 2000-2002. AB - In June 2002, a physician reported to the Oregon Department of Human Services (DHS) a case of acute hepatitis C in a patient who had received a patellar tendon with bone allograft from a donor approximately 6 weeks before onset of illness. At the time of the donor's death in October 2000, his serum had no detectable antibody to hepatitis C virus (anti-HCV). The ensuing investigation conducted by CDC and DHS confirmed that the donor, although anti-HCV-negative, was HCV RNA positive and the probable source of HCV infection for at least eight organ and tissue recipients. This report summarizes the preliminary results of the investigation. Although transmission from anti-HCV-negative tissue donors probably is rare, determining the frequency of transplantations from such donors and the risk for transmitting HCV to recipients is important in evaluating whether additional prevention measures are warranted. PMID- 12729078 TI - Supplemental recommendations on adverse events following smallpox vaccine in the pre-event vaccination program: recommendations of the Advisory Committee on Immunization Practices. AB - The Advisory Committee on Immunization Practices (ACIP) has issued recommendations previously for use of smallpox vaccine and supplemental recommendations for use of smallpox vaccine in the pre-event civilian vaccination program. On March 28, 2003, CDC reported cases of cardiac adverse events among persons vaccinated recently with smallpox vaccine. In response to these reports, ACIP held an emergency meeting on March 28 to make recommendations to CDC about medical screening of potential vaccinees and follow-up of persons with cardiovascular risk factors after vaccination. These recommendations supplement those previously issued by ACIP. PMID- 12729077 TI - Update: adverse events following smallpox vaccination--United States, 2003. AB - During January 24-March 28, 2003, smallpox vaccine was administered to 29,584 civilian health-care and public health workers in 54 jurisdictions as part of an effort to prepare the United States for a terrorist attack using smallpox virus. This report summarizes data on ten cases of cardiac adverse events reported among civilian vaccinees since the beginning of the smallpox vaccination program, including three new cardiac adverse events reported to CDC from the Vaccine Adverse a fall Event Reporting System (VAERS) during March 24-30. Fourteen cases of myocarditis and one fatal myocardial infarction have been reported among military personnel. This report summarizes data on the three new cases of cardiac adverse events, updates data on seven previously reported cases among civilian vaccinees (Table 1), summarizes selected cases of cardiac adverse events among military vaccinees, and updates information on all adverse events reported in the civilian vaccination program as of March 30. PMID- 12729079 TI - Biomimetic recognition and immunochemical assay of ligand binding to collagen. AB - PURPOSE: The objectives of this investigation were to analyze the adhesive interactions between type 1 collagen and two hydrophilic monomer primers (ligands) for dentin bonding through computer modeling and a novel immunochemical binding assay method. MATERIALS AND METHODS: The hydrophilic monomers studied included 2-hydroxyethyl methacrylate (HEMA) and 2-acryloyloxyethyl phosphate (PA). For computer modeling studies, a triple helical model structure of collagen fibril extracted from a public domain Protein Data Bank was used. The ligand conformations were modeled and optimized by Sybyl, and their interactions with the triple helical collagen structure in a solvent environment of water were simulated by AutoDock software. The effect of ligand binding to subsequent monoclonal antibody binding was also studied using an immunochemical binding assay method developed by us. The computer simulation results and binding assay results were analyzed for relationships pertinent to collagen-ligand interaction in bonding of dentin primers to demineralized dentin. RESULTS: Computer docking results indicate that ligand binding to collagen is favored to occur at cavity sites on the collagen molecular surfaces, where steric and electrostatic effects may play a critical role in mediating van der Waals and Coulombic interactions between ligand and receptor molecules. Prior ligand exposure of collagen reduces subsequent antibody binding during in vitro experiments. Differences in antibody binding were observed both as a function of ligand type and its concentration. CONCLUSION: Both steric complementarity and electrostatic complementarity conditions were observed in the docking site selection. Under conditions of ligand binding, antibody binding is diminished as a function of ligand structure and its concentration. The results suggest that approaches combining computer modeling and in vitro binding assay methods are powerful tools in evaluating dentinal adhesion at the atomic and/or molecular level. PMID- 12729080 TI - The effect of concentration and pH of silver nitrate solution on nanoleakage. AB - PURPOSE: The purpose of this study was to evaluate the effects of concentration and pH of silver nitrate solution on nanoleakage in occlusal flat surfaces bonded with three dentin bonding systems. MATERIALS AND METHODS: The dentin bonding systems used in this study were Single Bond, PermaQuik, and Prompt L-Pop. Flat occlusal dentin surfaces from extracted human molars were bonded with one of the dentin bonding systems. The teeth from each subgroup (n = 4) were placed in one of the following silver nitrate solutions: 50% w/v, 25% w/v, 10% w/v, and 2% w/v (pH 3.4, 4.2, 4.4, and 4.9, resp), and 50% w/v buffered to pH 6 in total darkness for 24 h, rinsed in running water for 5 min, immersed in photodeveloping solution for 24 h, and exposed to light for 8 h. Specimens were sectioned, mounted on stubs, carbon coated and observed in a Field Emission SEM using backscattered electron mode. RESULTS: FE-SEM images showed that samples from different concentrations of silver nitrate solution had leakage patterns similar to that of samples from 50% w/v for all dentin bonding systems. The concentrations and pH values of silver nitrate solutions did not affect leakage patterns in any of the systems. CONCLUSION: A lower concentration of silver nitrate solution may be used in a nanoleakage study, although the present results need to be replicated to determine an ideal concentration. PMID- 12729082 TI - Short- and long-term bonding efficacy of a self-etching, one-step adhesive. AB - PURPOSE: It was the purpose of the study to investigate the short- and long-term bonding to enamel and dentin of a self-etching, one-step adhesive with and without separate curing of the adhesive. The bond strengths were measured both with a light- and with a self-curing resin composite. Multistep bonding systems served as controls. MATERIALS AND METHODS: The bonding systems selected were Prompt L-Pop and, as controls, Solobond M, Solobond Plus, and Optibond FL. The resin composites Rebilda LC and Rebilda were bonded to plane enamel or dentin surfaces following application of the bonding systems used in accordance with the recommendations of the respective manufacturers. With Prompt L-Pop, a separate cure of the adhesive was also investigated. After being stored in water at 37 degrees C for 1 day or 1 year, the bonded specimens were broken in shear. RESULTS: The strength of the bond to enamel of Prompt L-Pop increased with time in water, while the bond strengths of the other systems did not change statistically significantly. There was no difference in bond strengths to dentin measured after 1 day and 1 year in water. The self-curing resin composite did not bond with Prompt L-Pop. Separate curing of Prompt L-Pop had no influence on bond strengths. CONCLUSION: On the basis of this in vitro study, the self-etching, one step bonding system Prompt L-Pop, although less efficient than the three-step system Optibond FL, would seem to be a viable option compared to multistep systems, except when there is a need to mediate a bond to self-curing resin composite. PMID- 12729081 TI - Factors contributing to the incompatibility between simplified-step adhesives and chemically-cured or dual-cured composites. Part I. Single-step self-etching adhesive. AB - PURPOSE: To determine if adverse chemical interaction and adhesive permeability are both responsible for the incompatibility between a single-step, self-etching adhesive and chemically-cured or dual-cured composites. MATERIALS AND METHODS: Bonding was performed with Xeno CF Bond (Dentsply-Sankin), on either hydrated (H) or dehydrated (DH) human dentin. For microtensile bond strength evaluation, a dual-cured hybrid composite (Bis-Core) was activated using: (1) the light-cured (L) mode (base syringe only), (2) delayed light activation (DL) (base syringe left on top of cured adhesive in the dark for 20 min before activation), and (3) the chemically-cured (C) mode (base and catalyst syringes in the dark). A chemical co-initiator (B; BondLink) was also applied to the cured adhesive before coupling with the composite in chemically-cured mode. This resulted in seven experimental groups: (1) L-H (control); (2) DL-H; (3) DL-DH; (4) C-H; (5) C-DH; (6) C-B-H; and (7) C-B-DH. For transmission electron microscopy, the dual-cured composite in the seven groups was replaced with a light-cured microfilled composite (Metafil CX) and an experimental chemically-cured microfilled composite of the same composition. Specimens were immersed in ammoniacal silver nitrate for 24 h. After reduction of the diamine silver ions to silver, undemineralized and unstained sections were examined for nanoleakage within the resin-dentin interfaces of the seven groups. RESULTS: For the light-cured modes, bond strengths fell substantially in DL-H but not in DL-DH. For the chemically-cured modes, bond strengths were lowest in C-H and only increased slightly in C-DH. The use of a chemical co-initiator with the adhesive further improved the bond strength in C-B-H. Only C-B-DH was not significantly different from the control light-cured mode L-H. Two abnormal modes of silver deposition were observed in resin-dentin interfaces. A continuous layer of silver was observed when the chemically-cured composite was applied to the cured adhesive in the absence of the chemical co-initiator (C-H; C-DH). Silver-impregnated water blisters were identified when the chemically-cured composite was coupled to bonded hydrated dentin (C-H; C-B-H). Similar water blisters were seen in DL-H in which adverse chemical interaction should not occur. CONCLUSION: Adverse chemical interaction between catalytic components of chemically-cured composite and the tested single step, self-etching adhesive was the major cause of reductions in bond strength, while adhesive permeability was a minor cause of bond strength reduction. The combination of these two factors accounts for the substantial reduction in bond strength when chemically-cured or dual-cured composites were coupled to bonded hydrated dentin. PMID- 12729083 TI - Microtensile bond strength of a total-etch 3-step, total-etch 2-step, self-etch 2 step, and a self-etch 1-step dentin bonding system through 15-month water storage. AB - PURPOSE: To evaluate dentin bond durability using current dentin adhesive resin bonding approaches over a 15-month period of water storage. MATERIALS AND METHODS: Forty-four extracted human molars were polished with 600-grit SiC papers exposing occlusal dentin, and randomly distributed into four adhesive groups: total-etch 3-step (TE3) (Scotchbond Multi-Purpose, 3M ESPE), total-etch 2-step (TE2) (Single Bond, 3M ESPE), self-etch 2-step (SE2) (Clearfil SE Bond, Kuraray), and a self-etch 1-step (SE1) (Prompt L-Pop, 3M ESPE). A resin composite crown was incrementally formed and light cured to approximately 6 mm in height. Microtensile specimens were fabricated and stored in distilled water containing 0.5% chloramine T and tensile tested at 1 mm/min after 1, 6, and 15 months. The debond pathway was recorded as either involving the substrate or joint using scanning electron microscopy. SAS software was used to compute Weibull parameters and distributions, Log-rank and Wilcoxon tests were used for comparison of survival curves over time for each adhesive system and between adhesive systems. RESULTS: The TE2 was significantly weaker than TE3 and SE2 after 1 and 6 months of storage, but all three systems were equivalent after 15 months of storage. The SE1 system could not be tested due to 58 of 65 specimens failing during specimen preparation. Failure modes were observed to be dependent upon adhesive system, with only the total-etch 2-step system demonstrating an increasing involvement in the adhesive joint over time. CONCLUSION: Although differences in bond strength were observed across adhesive systems up to 6 months of storage, no differences were noted at 15 months. This may represent common degradative mechanisms. PMID- 12729084 TI - Bond strength of glass-ionomer cements to caries-affected dentin. AB - PURPOSE: The aim of this study was to assess the tensile bond strength of a resin modified glass-ionomer cement (a: Fuji II LC) and three traditional glass-ionomer cements (b: Ketac-fil; c: Ketac Molar; d: Fuji IX) to caries-affected dentin. MATERIALS AND METHODS: Forty human permanent molars with occlusal caries in dentin were selected, embedded in polyester resin, and ground until the carious dentin was exposed. Infected dentin was removed with curettes according to the atraumatic restorative technique (ART), and the tooth surface was smoothed with SiC paper. A bonding site, limited to 3 mm in diameter, was treated with polyacrylic acid for 10 s. After surface treatment, an inverted glass-ionomer cone was prepared for each specimen, using a split bisected Teflon matrix. The cones were immediately protected with a thin layer of nail varnish or bonding agent. Specimens were stored in distilled water at 37 degrees C for 24 h, and then bond strength to failure was tested. RESULTS: The mean (SD) bond strengths in MPa were: a: 8.33 (2.35); b: 2.46 (1.60); c: 0.83 (1.18), and d: 1.45 (1.70). The data were analyzed using ANOVA and Tukey statistical tests. Fuji II LC, a resin-modified glass-ionomer cement, showed higher bond strength values and was statistically superior to the other groups, containing traditional glass ionomer cements (p < 0.05). CONCLUSION: Findings showed that the traditional glass ionomer cements tested in this study had lower mean bond strength values to caries-affected dentin than did the resin-modified glass-ionomer cement. PMID- 12729085 TI - Microleakage in ceramic inlays luted with different resin cements. AB - PURPOSE: The objective of this in vitro study was to evaluate the microleakage in ceramic inlays using different resin cements with margins in enamel and cementum/dentin interfaces. MATERIALS AND METHODS: Standard Class II MOD inlay cavities were prepared in 32 noncarious human premolars. The cavities were randomly divided into 4 groups (n = 8): CONTROL GROUP: cavities were treated with Single Bond and incrementally filled with a composite resin (P60); Enforce group: feldspathic ceramic inlays were luted using Prime & Bond 2.1 and Enforce; RelyX group: inlays were cemented with Single Bond and RelyX ARC; Resin Cement group: ceramic inlays were bonded using Single Bond and Resin Cement. Ceramic inlays were previously treated with 10% hydrofluoric acid for 2 min, followed by silane application. After 7 days of storage in distilled water, teeth were submitted to thermocycling. After applying nail varnish, specimens were immersed in 2% aqueous solution of methylene blue for 8 h. After washing, teeth were cut into three sections through the restorations, and the leakage was assessed using a standardized score. RESULTS: Data were submitted to statistical analysis using nonparametric tests (Mann-Whitney and Kruskal-Wallis). Dye leakage at margins in enamel was statistically lower (p < 0.01) than at cementum/dentin interfaces. RelyX ARC performed better (p < 0.05) than resin cement (enamel) and composite restorations (cementum/dentin). No other statistical differences were observed. CONCLUSIONS: Both the material and the substrate interface influenced microleakage of the ceramic inlays. PMID- 12729087 TI - Psyche and soma. PMID- 12729086 TI - Contraction stress of composite resin build-up procedures for pulpless molars. AB - PURPOSE: The purpose of this study was to photoelastically evaluate contraction stresses associated with various resin composite build-up procedures for pulpless molars. MATERIALS AND METHODS: Photoelastic models of endodontically treated mandibular molars were fabricated to simulate a preparation for a full-cast crown. The model configuration included three lateral walls, but no post space. The buildups were made with dual-curing resin composite using the following procedures: 1) bulk dual cure, 2) bulk chemical cure, 3) horizontal two increments, dual cure, and 4) indirect. Five specimens were fabricated for each condition. The stresses developed in the models were recorded photographically in the field of a circular polariscope. RESULTS: The build-up procedures tested generated widely different stress distributions and intensities. The highest stresses were seen with the bulk dual-cure method. The slower polymerization chemical-cure group developed distributions similar to the bulk dual-cure group with a significantly lower fringe order. The two horizontal increments dual-cure techniques developed an individual group of fringes for each layer. Compared with the bulk dual-cure group, stresses around the occlusal margin were reduced by incrementalization, while fringes were more closely spaced at the line angles with a slightly lower fringe order. The indirect method demonstrated the lowest stress which extended over the smallest area. CONCLUSION: Contraction stress in resin composite buildups varied significantly depending upon the procedures of fabrication. The bulk dual-cure method developed the most severe contraction stresses, while the indirect technique resulted in significantly lower contraction stresses than the other techniques tested. PMID- 12729088 TI - What's eating you? Enterobius vermicularis (pinworms, threadworms). PMID- 12729089 TI - Epidemiology of skin diseases in people of color. AB - The epidemiology of skin diseases in people of color has not been extensively studied. Many skin diseases (eg, acne vulgaris; eczematous dermatitis; infections caused by bacteria, fungi, or viruses) are common to most people of color- blacks, Asians, Hispanics/Latinos, and Native Americans. Diseases of more cosmetic concern (eg, melasma, postinflammatory pigmentation, acne keloidalis nuchae, scalp and facial folliculitis, keloids, alopecias) occur more in skin of color than in white skin. PMID- 12729090 TI - Primary systemic amyloidosis. PMID- 12729091 TI - Cobb syndrome associated with a verrucous (angiokeratomalike) vascular malformation. AB - Cobb syndrome is defined by a cutaneous vascular lesion and a corresponding spinal cord vascular malformation within a segment or two of the involved dermatome. Even though Cobb syndrome has been reported in association with various cutaneous vascular lesions, to our knowledge, only one other previous report describes verrucous changes overlying the cutaneous vascular lesion. We describe a patient with verrucous vascular malformation extending from his right chest to his right upper back in a dermatomal distribution. The patient's medical history included excision of a cavernous vascular malformation from the corresponding segment of his thoracic spinal cord 4 years prior, reversing 7 years of progressive leg weakness and neural deficits. Cobb syndrome was diagnosed based on the dermatomally distributed vascular malformation and the corresponding spinal cord vascular malformation. His skin lesions initially improved with erbium: YAG laser treatment but later recurred. This is the second reported case of Cobb syndrome associated with verrucous angiokeratomalike changes overlying the cutaneous vascular malformation. Because of the potentially severe neurologic sequelae caused by spinal cord lesions, clinicians should evaluate patients with dermatomally distributed cutaneous lesions on the trunk or extremities, including vascular malformations with verrucous features. PMID- 12729092 TI - A case of human orf contracted from a deer. AB - Orf, or contagious ecthyma, is a rare viral dermatosis caused by a member of the genus Parapoxvirus. The typical lesion consists of solitary or multiple papules that progress through a series of stages, terminating in complete resolution. This zoonotic disease is most commonly transmitted to humans from infected sheep or goats. We report a case of human orf, likely contracted from exposure to deer carcasses. PMID- 12729093 TI - Picking apart the picker: a clinician's guide for management of the patient presenting with excoriations. AB - Management of patients who "pick" at their skin is often difficult. Etiologies and maintaining factors can be unclear. Significant psychiatric overlay is often present, and the clinician is left with an overwhelming differential diagnosis and a poorly focused treatment plan that can result in suboptimal clinical outcomes. The purpose of this paper is to provide a conceptual framework for the evaluation and treatment of patients presenting with excoriations. Ten diagnostic categories are examined, and specific suggestions for treatment are offered. PMID- 12729094 TI - Trachyonychia: a case report and review of manifestations, associations, and treatments. AB - Trachyonychia ("rough nails") is best considered a reaction or morphologic pattern with a variety of clinical presentations and etiologies. It may involve only 1 or as many as 20 nails (20-nail dystrophy). It can be a manifestation of lichen planus, psoriasis, alopecia areata, immunoglobulin A deficiency, atopic dermatitis, and ichthyosis vulgaris. Nail matrix biopsy results and physical examination findings help in establishing the cause of this condition, though often trachyonychia is an isolated finding. When trachyonychia occurs in childhood as a manifestation of lichen planus, it tends to resolve with time. We review a case of trachyonychia, its association, its diagnostic evaluation, and treatment options. PMID- 12729095 TI - Pemphigus erythematosus associated with anti-DNA antibodies and multiple anti-ENA antibodies: a case report. AB - Pemphigus erythematosus (PE) is an autoimmune blistering disease combining features of pemphigus foliaceus (PF) and systemic lupus erythematosus (SLE). We report a case of PE associated with anti-double-stranded DNA (anti-dsDNA), anti Smith (anti-Sm), anti-Ro (anti-SSA), and antiribonucleoprotein (anti-RNP) antibodies. This case required extensive immunosuppressive therapy. We treated our patient with a combination of systemic steroids, intramuscular gold injections, azathioprine, and hydroxychloroquine. The patient's response was complete remission--evaluated clinically, serologically, and immunohistochemically. PMID- 12729096 TI - Betel quid-induced oral lichen planus: a case report. AB - The social use of betel nut is relatively common in certain geographic areas, especially India and Southeast Asia. The term betel nut does not truly describe the product that is chewed; rather, the term quid is more accurate because it refers to a substance or mixture of substances, including the areca nut, that are chewed and remain in contact with the mucosa. Betel quid is a type of quid that contains betel leaf. Chewer's mucosa and oral submucous fibrosis are clinical entities that have been associated with betel quid use. We report a case of oral lichen planus induced by betel quid use in a 79-year-old Cambodian woman. PMID- 12729097 TI - Mid-dermal elastolysis. AB - Mid-dermal elastolysis (MDE), which presents as fine wrinkling of the skin or perifollicular papules, is extremely rare. This entity is distinguished from other elastolytic disorders by its characteristic bandlike loss of elastic fibers limited to the mid dermis. We report a case of MDE that developed gradually in an otherwise healthy woman without prior cutaneous disease. The current theories on the pathogenesis of MDE also are discussed. PMID- 12729098 TI - Blue rubber-bleb nevus syndrome: a case report. AB - Blue rubber-bleb nevus syndrome (BRBNS) is an uncommon systemic disorder characterized by cutaneous and visceral cavernous hemangiomas. The characteristic rubbery textured and easily compressible lesions usually present in childhood and predominate cutaneously over the trunk and extremities. Gastrointestinal foci appear most commonly in the small bowel, a site that appears to dominate visceral involvement. We review the case of a 23-year-old white woman, whose numerous lesions on her trunk, extremities, and oral mucosa had been present since childhood. PMID- 12729099 TI - Smallpox. AB - With recent events, the threat of bioterrorism has become a reality. In late 2001, multiple cases of cutaneous and inhalation anthrax were spread through the US mail. On the front line were dermatologists who diagnosed the first cases of cutaneous anthrax in New York City. Since then, physicians who are unsure if they are facing a new form of bioterrorism frequently have consulted dermatologists to evaluate rashes. Because most biological weapons (anthrax, tularemia, plague, smallpox) can have cutaneous manifestations, dermatologists will continue to have an important role in evaluating these potential threats. PMID- 12729100 TI - Spontaneous anterior interosseous nerve palsy. PMID- 12729101 TI - Computer-assisted orthopaedic surgery: do we need CAOS? PMID- 12729102 TI - Diagnosis of a tear of the tibiofibular syndesmosis. The role of arthroscopy of the ankle. AB - In 52 patients we compared the accuracy of standard anteroposterior (AP) radiography, mortise radiography and MRI with arthroscopy of the ankle for the diagnosis of a tear of the tibiofibular syndesmosis. In comparison with arthroscopy, the sensitivity, specificity and accuracy were 44.1%, 100% and 63.5% for standard AP radiography and 58.3%, 100% and 71.2% for mortise radiography. For MRI they were 100%, 93.1% and 96.2% for a tear of the anterior inferior tibiofibular ligament and 100%, 100% and 100% for a tear of the posterior inferior tibiofibular ligament. Standard AP and mortise radiography did not always provide a correct diagnosis. MRI was useful although there were two-false positive cases. We suggest that arthroscopy of the ankle is indispensable for the accurate diagnosis of a tear of the tibiofibular syndesmosis. PMID- 12729103 TI - Synovial impingement in the ankle. A new physical sign. AB - Inversion injuries of the ankle are common and most are managed adequately by functional treatment. A significant number will, however, remain symptomatic. Synovial impingement is one cause of continuing pain. This condition is often difficult to diagnose because the physical signs and investigations are non specific. If the diagnosis is made, treatment by arthroscopic debridement has been shown to be highly effective. Our aim was to describe a new physical sign to help in the diagnosis of anterolateral synovial impingement in the ankle. A cadaver dissection demonstrated the anatomical basis for the physical sign and a prospective clinical study involving 73 patients showed that the lateral synovial impingement test had a sensitivity of 94.8% and a specificity of 88%. We describe the test and conclude that this physical sign will be of use to practitioners treating patients with chronic pain in the ankle after injury. PMID- 12729104 TI - Total ankle replacement. The results in 200 ankles. AB - Between 1993 and 2000 we implanted 200 cementless, mobile-bearing STAR total ankle replacements. None was lost to follow-up for reasons other than the death of a patient. The mean follow-up was for 46 months (24 to 101). A complication requiring further surgery developed in eight ankles and 14 were revised or fused. The cumulative survival rate at five years was 92.7% (95% CI 86.6 to 98.8) with time to decision to revision or fusion as an endpoint. The most frequent complications were delayed wound healing and fracture of a malleolus. These became less common with experience of the operation. The radiological appearance of the interface of the tibial implant was significantly related to its operative fit and to the type of bioactive coating. PMID- 12729105 TI - The treatment of recurrent posterolateral instability of the elbow. AB - This study describes the surgical technique used for reconstruction and reinforcement of the lateral collateral ligament complex in patients with posterolateral instability of the elbow and the results. A triceps tendon graft from the ipsilateral elbow which was inserted through bone tunnels and fixed with bone anchors augmented the reconstruction. The operation was performed on 18 consecutive patients with instability after an acute traumatic dislocation. The mean follow-up was 44 months (14 to 88). There were no recurrent dislocations. The elbow was stable in 14 patients; three had some minor limitation of movement. Thirteen had no or only occasional slight pain, 15 returned to their normal level of activity and 17 were satisfied with the outcome. There was only one failure. PMID- 12729106 TI - Treatment of primary degenerative arthritis of the elbow by ulnohumeral arthroplasty. A long-term follow-up. AB - Between 1990 and 1996 we performed 20 consecutive ulnohumeral arthroplasties for primary osteoarthritis of the elbow. The outcome was assessed using the Disabilities of Arm, Shoulder and Hand Score (DASH) and the Mayo Elbow Performance Score (MEPS) at a mean follow-up of 75 months (58 to 132). There were excellent or good results in 17 elbows (85%) using the DASH score and in 13 (65%) with the MEPS (correlation coefficient 0.79). The mean fixed flexion deformity had improved by 10 degrees and the range of flexion by a mean of 20 degrees. In 16 elbows (80%) the benefits of surgery had been maintained, and of 16 patients working at the time of operation, 12 (75%) had returned to the same job. There was no correlation between radiological recurrence of degenerative changes and the amount of fixed flexion deformity, the flexion arc, or the elbow scores. PMID- 12729107 TI - Early results of the Souter-Strathclyde unlinked total elbow arthroplasty in patients with osteoarthritis. AB - We have carried out a retrospective review of 11 Souter-Strathclyde primary total elbow arthroplasties in ten patients with osteoarthritis, over a period of nine years. The diagnosis was primary osteoarthritis in nine elbows and post-traumatic arthritis in two. The mean follow-up was 68 months (15 to 117). Although no patient was symptomatic, radiological review revealed evidence of loosening affecting three humeral and two ulnar components, one of which subsequently failed and was revised at 97 months. There were no dislocations, deep infections or mechanical failures. Complications included two superficial wound infections and two neurapraxias of the ulnar nerve which resolved. This study shows that the unlinked Souter-Strathclyde total elbow arthroplasty can be considered for patients with osteoarthritis and gives good symptomatic relief and improvement in function. PMID- 12729108 TI - Total elbow replacement using the Kudo prosthesis. Clinical and radiological review with five- to seven-year follow-up. AB - Between 1993 and 1996, we undertook 35 Kudo 5 total elbow replacements in a consecutive series of 31 rheumatoid patients. A total of 25 patients (29 procedures) was evaluated at a mean follow-up of six years (5 to 7.5) using the Mayo Clinic performance index. In addition, all patients were assessed for loosening using standard anteroposterior and lateral radiographs. At review, 19 elbows (65%) had either no pain or mild pain, ten (35%) had moderate pain and none had severe pain. The mean arc of flexion/extension was 94 degrees (35 to 130) and supination/pronation was 128 degrees (30 to 165). A fracture of the medial epicondyle occurred during surgery in one patient. This was successfully treated with a single AO screw and a standard Kudo 5 implant was inserted. Postoperatively, there were no infections. One patient had a dislocation which was treated by closed reduction and five had neurapraxia of the ulnar nerve. Radiologically, there was no evidence of loosening of the humeral component, but two ulnar components had progressive radiolucent lines suggestive of loosening. Two other ulnar components had incomplete and non-progressive radiolucent lines. With definite radiological loosening as the endpoint, the probability of survival of the Kudo 5 prosthesis at five years using the Kaplan-Meier method was 89%. PMID- 12729109 TI - Radiographic observation of the scaphoid shift test. AB - The movements of the carpal bones during the scaphoid shift test were evaluated radiographically in 60 wrists. The clinical results were graded according to the degree of subluxation of the scaphoid and pain on the dorsum of the wrist. Lateral radiographs at rest and under stress were taken and the relative movements of the scaphoid with respect to the radius and lunate, and the rotation of the scaphoid and lunate were calculated. Dorsal displacement of the scaphoid with respect to the radius was significantly associated with the clinical grade of subluxation. There was correlation between the amount of pain and the displacement of the scaphoid from the lunate, but not from the radius. The wrists with a painful shift test had greater relative displacement of the scaphoid from the lunate than those with painless subluxation. These observations support the view that pain associated with subluxation of the scaphoid during the shift test is a significant finding, and that radiographic analysis of the test may confirm a positive result when dynamic scaphoid instability is suspected. PMID- 12729110 TI - Strain on the human sciatic nerve in vivo during movement of the hip and knee. AB - Injuries to the sciatic nerve are an occasional complication of surgery to the hip and acetabulum, and traction is frequently the causative mechanism. In vitro and animal experiments have shown that increased tensile strain on peripheral nerves, when applied for prolonged periods, impairs nerve function. We have used video-extensometry to measure strain on the human sciatic nerve during total hip replacement (THR). Ten consecutive patients with a mean age of 72 years undergoing primary THR by the posterior approach were recruited, and strains in the sciatic nerve were measured in different combinations of flexion and extension of the hip and knee, before dislocation of the hip. Significant increases (p = 0.02) in strain in the sciatic nerve were observed in flexion of the hip and extension of the knee. The mean increase was 26% (19% to 30%). In animal studies increases of this magnitude have been shown to impair electrophysiological function in peripheral nerves. Our results suggest that excessive flexion of the hip and extension of the knee should be avoided during THR. PMID- 12729111 TI - A femoral component with proximal HA coating. An analysis of survival and fixation at up to ten years. AB - We describe the survival at ten years of 100 femoral components of the Freeman hip prosthesis. It is proximally hydroxyapatite (HA)-coated and was fixed without cement. Radiological assessment identified radiolucent lines (RLLs) and lytic lesions and was used to measure migration. The criterion of failure was revision or impending revision for aseptic femoral loosening. No femoral components were revised or are awaiting revision for aseptic loosening, giving 100% survival at ten years (95% confidence interval, 95.7 to 100), although 59 were at risk at ten years. Two components were revised for fracture of a ceramic head with damage to the trunnion. Although well fixed in each, for survival analysis we evaluated the hip as if the patient had died. Twelve acetabular components were revised and at each operation the femoral component was found to be well fixed, was not disturbed and remained in the survival analysis. Three patients were lost to follow-up, and 12 died with well-functioning prostheses. Radiologically, all except one of the components appeared to be well fixed with no RLLs and no lytic lesions at the latest follow-up. The mean vertical migration was 0.4 mm at one year, 0.8 mm at two years and 1.4 mm at ten years. One component had migrated 7.6 mm at ten years (2.1 mm in year 1) and developed RLLs in Gruen zones I and II. The symptoms, however, were only minor and revision was not indicated. Our study has shown that proximal HA coating gives effective fixation for a femoral component. PMID- 12729112 TI - Hyperbaric oxygen therapy as a treatment for stage-I avascular necrosis of the femoral head. AB - Avascular necrosis (AVN) of the head of the femur is a potentially crippling disease which mainly affects young adults. Although treatment by exposure to hyperbaric oxygen (HBO) is reported as being beneficial, there has been no study of its use in treated compared with untreated patients. We selected 12 patients who suffered from Steinberg stage-I AVN of the head of the femur (four bilateral) whose lesions were 4 mm or more thick and/or 12.5 mm or more long on MRI. Daily HBO therapy was given for 100 days to each patient. All smaller stage-I lesions and more advanced stages of AVN were excluded. These size criteria were chosen in order to compare outcomes with an identical size of lesion in an untreated group described earlier. Overall, 81% of patients who received HBO therapy showed a return to normal on MRI as compared with 17% in the untreated group. We therefore conclude that hyperbaric oxygen is effective in the treatment of stage-I AVN of the head of the femur. PMID- 12729113 TI - Wear of enhanced ultra-high molecular-weight polyethylene (Hylamer) in combination with a 22.225 mm diameter zirconia femoral head. AB - We have prospectively studied the wear of enhanced ultra-high molecular-weight polyethylene (Hylamer) in combination with a zirconia femoral head of 22.225 mm diameter on a cemented, triple-tapered, collarless, polished stem, the C Stem. The 71 patients who underwent total hip arthroplasty had a mean follow-up of six years (3 to 8). No patient died or was lost to follow-up. The clinical results were excellent with 96% of patients satisfied. There were no revisions. Two cups were considered to be loose radiologically. One was avulsed from the cement in a skiing accident, with a periprosthetic fracture of the femur, but has remained stable for more than seven years. One femur shows radiological appearances which are compatible with a healing infection. One femoral component is at risk of loosening. The mean rate of penetration of the cup was 0.22 mm/year (0.06 to 0.55). Our results appear to be within the guidelines set by the National Institute of Clinical Excellence. We have discontinued the use of Hylamer despite excellent clinical results and no revisions to date because the high initial rates of penetration did not settle to the expected low levels within the anticipated time. PMID- 12729114 TI - Internal fixation compared with total hip replacement for displaced femoral neck fractures in the elderly. A randomised, controlled trial. AB - The treatment algorithms for displaced fractures of the femoral neck need to be improved if we are to reduce the need for secondary surgery. We have studied 102 patients of mean age 80 years, with an acute displaced fracture of the femoral neck. They were randomly placed into two groups, treated either by internal fixation (IF) with two cannulated screws or total hip replacement (THR). None showed severe cognitive dysfunction, all were able to walk independently, and all lived in their own home. They were reviewed at four, 12 and 24 months after surgery. Outcome measurements included hip complications, revision surgery, hip function according to Charnley and the health-related quality of life (HRQoL) according to EuroQol (EQ-5D). The failure rate after 24 months was higher in the IF group than in the THR group with regard to hip complications (36% and 4%, respectively; p < 0.001), and the number of revision procedures (42% and 4%, p < 0.001). Hip function was significantly better in the THR group at all follow-up reviews regarding pain (p < 0.005), movement (p < 0.05 except at 4 months) and walking (p < 0.05). The reduction in HRQoL (EQ-5D index score) was also significantly lower in the THR group than in the IF group, comparing the pre fracture situation with that at all follow-up reviews (p < 0.05). The results of our study strongly suggest that THR provides a better outcome than IF for elderly, relatively healthy, lucid patients with a displaced fracture of the femoral neck. PMID- 12729115 TI - The role of the posterior cruciate ligament in total knee replacement. AB - We randomised 129 knees which were to be replaced using a standard posterior cruciate-ligament (PCL)-retaining cemented total knee replacement into two groups. In one the PCL was retained in the normal way and in the other it was resected. They were well matched, with a predominance of women, and a mean age of 67 years. There was no statistically significant difference in the Hospital for Special Surgery scores at a mean of 57 months (56 to 60) between the two groups although 21 patients (24 knees) were lost to follow-up. Relief from pain, correction of deformity, range of movement, stability and strength were comparable in both. Radiological assessment showed femoral rollback in approximately 20% of knees with a slightly higher incidence in the PCL-resected group. There was no significant loosening detected in either group at review at two years. At five years, one knee in the PCL-retained group had been revised because of infection and one patient in each group was awaiting revision for loosening. Our findings have shown no significant difference in the five-year results for a PCL-retaining total knee replacement if the PCL is excised or preserved. This suggests two important points. First, the PCL is not functional in most patients with a total knee replacement even when retained. Secondly, patients with an excised PCL show a good result with a PCL-retaining implant, thereby questioning the need for a posterior stabilised design in such a situation. PMID- 12729116 TI - Role of abrasion of the femoral component in revision knee arthroplasty. AB - We carried out 60 revision procedures for failed porous coated anatomic total knee replacements in 54 patients, which were divided into two groups. The 14 knees in group I had a well-fixed femoral component at surgery which was retained, and in the 46 knees in group II both tibial and femoral components were loose and were revised using a variety of implants. Our review comprised clinical and radiological assessment. A total of 13 knees required a second revision. Six (42%) in group I failed very early (mean 2.1 years) when compared with seven (15%) in group II (mean 6.8 years). Failure was due to wear of the polyethylene insert by the abraded, retained femoral component (crude odds ratio 4.07; 95% CI 1.07 to 15.5). We recommend a complete change of primary bearing surfaces at the time of revision of an uncemented total knee replacement in order to prevent early wear of polyethylene. PMID- 12729117 TI - Vascular malformations of the lower limb with osseous involvement. AB - Vascular malformations are rare congenital lesions which often have associated skeletal changes. Over a period of ten years, 90 patients at our clinic had a vascular anomaly of the lower limb, examined by either CT or MRI. Of these, 18 (20%) had bony involvement. A questionnaire was sent to these patients (8 men, 10 women) to evaluate their age of presentation, initial symptoms and current complaints. Radiological imaging revealed 15 low- and three high-flow lesions. The mean age at presentation to a physician was six years of age. Pain was the most common complaint. Disparity in leg length of 2 cm or more was observed in ten patients. Of the 16 patients with muscle infiltration, 13 had four or more muscles involved. Treatment by resection alone would require radical surgery. PMID- 12729118 TI - The relationship between the strength of supination of the forearm and rotation of the shoulder. AB - In children with obstetric brachial plexus palsy (OBPP) who develop an internal rotation deformity of the shoulder, release of subscapularis improves the range of external rotation of the shoulder and the strength of supination of the forearm. We studied the strength of supination in 35 healthy adult volunteers at 45 degrees of both internal and external rotation. The mean and maximum torques were greater in external than internal rotation by 8.7% and 7.5%, respectively. This was highly significant (p < 0.0001). The increased strength of supination in external rotation is probably because the maximum power of biceps, particularly the long head, may be exerted in this position. In children the difference may be even greater due to anatomical differences causing the dramatic increases in the strength of supination after surgery for OBPP. In adults our findings suggest that the supination exercises which are undertaken after injury or surgery to the forearm or wrist should be performed in external rotation. PMID- 12729119 TI - Impingement syndrome associated with whiplash injury. AB - We examined 524 patients with whiplash injuries for delayed onset of shoulder pain in order to establish whether this was due to impingement syndrome. A total of 476 patients (91%) responded to a questionnaire of which 102 (22%) were entered into the study; 43 had both a positive impingement sign and Neer test. The incidence of impingement-type pain was 9%. After treatment 23 patients (5%) had a significant improvement in their symptoms, ten (2%) had a moderate improvement and nine had no improvement. Impingement-type pain can occur after whiplash injuries and can be successfully treated. PMID- 12729121 TI - Rehabilitation after two-part fractures of the neck of the humerus. AB - We undertook a prospective, controlled trial which compared two rehabilitation programmes for 86 patients who sustained two-part fractures of the proximal humerus. Patients were randomised either to receive immediate physiotherapy within one week (group A) or delayed physiotherapy after three weeks of immobilisation in a collar and cuff sling (group B). At 16 weeks after the fracture, patients in group A had less pain (p < 0.01) and had greater shoulder function (p < 0.001) than those in group B. At 52 weeks, the differences between the groups had reduced. Although group A still had greater shoulder function and less pain, there was no statistical difference when compared with group B. By analysis of the area under the curve, an overall measure up to the 52-week period, group A experienced less pain as measured by the SF36 general health questionnaire and had improved shoulder function. Our results show that patients with two-part fractures of the proximal humerus who begin immediate physiotherapy, experience less pain. The gains in shoulder function persist at 52 weeks which suggests that patients do not benefit from immobilisation before beginning physiotherapy. PMID- 12729120 TI - Iliosacral screw fixation of the posterior pelvic ring using local anaesthesia and computerised tomography. AB - Various techniques have been used for the fixation of the posterior pelvis, each with disadvantages specific to the technique. In this study, a new protocol involving the placement of posterior pelvic screws in the CT suite is described and evaluated. A total of 66 patients with unstable pelvic ring injuries was stabilised under local anaesthesia with sedation. The mean length of time for the procedure was 26 minutes per screw. There were no technical difficulties or misplaced screws and no cases of infection or nonunion. All patients stated that they would choose to have the CT scan procedure again rather than a procedure requiring general anaesthesia. The charges for the procedure were approximately 1840 pounds sterling (2800 dollars) per operation. CT-guided placement of iliosacral screws is a safe, feasible, and cost-effective alternative to radiologically-guided placement in the operating theatre in selected patients. PMID- 12729122 TI - Peripheral bone mineral density in patients with distal radial fractures. AB - Fractures of the distal forearm are widely regarded as the result of "fragility". We have examined the extent to which patients with Colles' fractures have osteopenia. We measured the bone mineral density (BMD) in the contralateral radius of 235 women presenting with Colles' fractures over a period of two years. While women of all ages had low values for ultra-distal BMD, the values, in age matched terms, were particularly low among premenopausal women aged less than 45 years. This result was not due to the presence of women with an early menopause. This large survey confirms and extends the findings from earlier small studies. We consider that it is particularly important to investigate young patients with fractures of the distal forearm to identify those with osteoporosis, to seek an underlying cause and to consider treatment. PMID- 12729123 TI - Fixation of fractures of the femoral neck. A prospective, randomised trial of three Ullevaal hip screws versus two Hansson hook-pins. AB - In a prospective, randomised trial, we compared the use of three Ullevaal hip screws with that of two Hansson hook-pins in 278 patients with fractures of the femoral neck. Background factors were similar in both groups. Follow-up was for two years. There were no significant differences between the groups in length of time of surgery, hospital stay, general complications, mortality, pain or walking ability. Likewise, the rates of early failure of fixation, nonunion, and the need for reoperation did not differ significantly between the groups. The use of hook pins was associated with less drill penetrations of the femoral head during surgery (odds ratio 2.6, p = 0.05) and a lower incidence of necrosis of the femoral head (odds ratio 3.5, p = 0.04). There was a strong relationship between poor reduction and fixation of the fracture and subsequent reoperation (p = 0.0005 and p = 0.0001, respectively). Likewise, peroperative drill penetration of the femoral head was associated with a greater risk of reoperation (p = 0.038). Both methods gave favourable results. In total, 22% of the patients needed a major reoperation (usually hemiarthroplasty), while in 7% of the cases the fixation device needed to be removed. Osteosynthesis as the sole method for operation of all fractures of the femoral neck was thus successful in 78% of patients. With selective treatment most of the remaining patients would have benefited if treated by a primary arthroplasty. Accurate selection requires the development of better prognostic methods. PMID- 12729124 TI - Extra-articular extrusion of an osteochondral fragment of the talar dome. AB - Osteochondral fractures of the talus are uncommon. They are classified according to Berndt and Harty, as progressing in severity through four stages. This classification, however, does not address extra-articular extrusion of the osteochondral fragment. We report an osteochondral lesion of the talar dome which presented as an extruded extra-articular fragment in a closed injury of the ankle. This type of lesion may offer a continuation to the four original stages. Clinicians should be aware that this pattern of fracture can occur, and thus allow a more accurate diagnosis and the provision of some aid in the treatment of these injuries. PMID- 12729125 TI - Contamination during removal of cement in revision hip arthroplasty. A cadaver study using ultrasound and high-speed cutters. AB - Instruments used in surgery which rotate or vibrate at a high frequency can produce potentially contaminated aerosols. Such tools are in use in cemented hip revision arthroplasties. We aimed to measure the extent of the environmental and body contamination caused by an ultrasound device and a high-speed cutter. On a human cadaver we carried out a complete surgical procedure including draping and simulated blood flow contaminated with Staphylococcus aureus (ATCC 12600). After cemented total hip arthroplasty, we undertook repeated extractions of cement using either an ultrasound device or a high-speed cutter. Surveillance cultures detected any environmental and body contamination of the surgical team. Environmental contamination was present in an area of 6 x 8 m for both devices. The concentration of contamination was lower for the ultrasound device. Both the ultrasound and the high-speed cutter contaminated all members of the surgical team. The devices tested produced aerosols which covered the whole operating theatre and all personnel present during the procedure. In contaminated and infected patients, infectious agents may be present in these aerosols. We therefore recommend the introduction of effective measures to control infection and thorough disinfection of the operating theatre after such procedures. PMID- 12729126 TI - Tissue reactions adjacent to well-fixed hydroxyapatite-coated acetabular cups. Histopathology of ten specimens retrieved at reoperation after 0.3 to 5.8 years. AB - Ten acetabular cups coated with hydroxyapatite (HA) had originally been inserted in five primary and five revision total hip replacements. The thickness of the HA was 155 +/- 35 microm. The cups, which were well-fixed, were retrieved, with their adherent tissue, at reoperation after 0.3 to 5.8 years because of infection (five hips), wear of polyethylene (three hips), and instability (two hips). Undecalcified sections showed a direct contact between bone and osteoid-like tissue which had formed directly onto the HA coating. The area within the threads and their mirror images, as well as the implant-tissue interfaces consisted of similar amounts of bone and soft tissue. Degradation of HA was seen in all hips. The mean thickness of the remaining HA coating was 97 microm (95% CI 94 to 101). The metal interface comprised 66% HA. The HA-tissue interface contained more bone than soft tissue (p = 0.001), whereas the metal-tissue interface included more soft tissue than bone (p = 0.019). Soft tissue at the implant interface and poor replacement of HA by bone may interfere with long-term fixation. PMID- 12729127 TI - Regulation of expression of alpha-smooth muscle actin in cells of Dupuytren's contracture. AB - Our aims were to describe the distribution of alpha-smooth muscle actin (SMA) containing cells in Dupuytren's tissue in vivo and to determine the effects of selected agents in regulating the expression of SMA in Dupuytren's cells in vitro. In selected hypercellular zones of Dupuytren's nodules up to 40% of the cells contained SMA, as shown by immunohistochemistry. A lower percentage (20%) of SMA-containing cells was found in regions of lower cellularity. A notable finding was that treatment in vitro of Dupuytren's cells with platelet-derived growth factor significantly reduced the content of SMA. Cells from the same patients showed a significant increase in expression of SMA in response to treatment with transforming growth factor, which confirmed recent findings. In addition, interferon-gamma, which has been previously used as a treatment for Dupuytren's disease in a clinical study, had no reproducible effect on the expression of this actin isoform. Our findings are of significance for the conservative management of contractures. PMID- 12729128 TI - Doxycycline inhibits bone resorption by human interface membrane cells from aseptically loose hip replacements. AB - Matrix metalloproteinases (MMPs) may have a role in the process of aseptic loosening. Doxycycline has been shown to inhibit MMPs. Our aim was to investigate the potential pharmacological effect of doxycycline on aseptic loosening. We used radiolabelled mouse calvariae cultured with human interface membrane cells from aseptically loosened hips. Bone resorption was confirmed in this model. The effect of doxycycline was assessed by culturing dead radiolabelled bone discs with cells from the interface membrane with doxycycline. The control group consisted of the same culture system without doxycycline. Supernatant 45calcium and the total 45calcium remaining in the bone discs at the completion of the culture were used to measure osteolysis. We found that doxycycline can inhibit osteolysis at the interface membrane of aseptically loosened hips. This may have therapeutic implications for the treatment of patients with aseptic loosening of total joint replacements. PMID- 12729129 TI - Early lesions of the labrum and acetabular cartilage in osteonecrosis of the femoral head. PMID- 12729130 TI - Management of completely displaced metaphyseal fractures of the distal radius in children. PMID- 12729131 TI - The provision of services for spinal disorders. PMID- 12729132 TI - Observer reliability in the arthroscopic classification of osteoarthritis of the knee. PMID- 12729133 TI - Simple treatment for torus fractures of the distal radius. PMID- 12729134 TI - A prospective, randomised trial of internal fixation versus arthroplasty for displaced fractures of the neck of the femur. PMID- 12729136 TI - Family planning providers' perspectives on dual protection. AB - CONTEXT: Family planning providers can play an important role in helping women to identify their risk of HIV and other sexually transmitted diseases (STDs) and to adopt preventive measures. In-depth investigation of providers' attitudes about approaches to STD risk assessment, contraceptive counseling and dual protection- concurrent protection from STDs and unintended pregnancy--has been limited. METHODS: In semistructured interviews conducted in 1998, 22 health care providers from a large New York City agency offering contraceptive and STD services described how they balanced STD and pregnancy concerns, viewed risk assessment and assessed various contraceptive methods. RESULTS: STD prevention was seen as an integral part of family planning counseling, and most providers believed that risk assessment should be conducted universally. Providers viewed dual protection as use of condoms along with an effective contraceptive; few advocated use of the male or female condom alone. The female condom was believed to be a disease prevention method of last resort and was considered appropriate only for specific groups of women. Although providers lacked understanding about the effectiveness of the female condom and how to counsel clients concerning its use, they expressed interest in learning more. CONCLUSIONS: Training is needed to reduce providers' negative perceptions of the female condom and to reinforce the importance of individualized counseling tailored to women's specific circumstances. Studies are needed on how to encourage family planning providers to promote male and female condoms as effective contraceptive methods. PMID- 12729135 TI - An evaluation of California's Adolescent Sibling Pregnancy Prevention Program. AB - CONTEXT: The siblings of adolescents who have been pregnant or are parents have disproportionately high rates of teenage pregnancies and births. California's Adolescent Sibling Pregnancy Prevention Program is targeted at these high-risk youths. METHODS: An evaluation of the program was conducted in 1997-1999 with 1,176 predominantly Hispanic 11-17-year-olds who had at least one sibling who was an adolescent parent or had been pregnant--731 youths who were program clients and 445 youths who received no systematic services. All evaluation participants completed an interview and questionnaire at enrollment and again nine months later. RESULTS: Female program clients had a significantly lower pregnancy rate than comparison females over the evaluation period (4% vs. 7%), as well as a lower rate of sexual initiation (7% vs. 16%). They also significantly decreased their frequency of school truancy, whereas this outcome increased among comparison females; program females had significantly more definite intentions of remaining abstinent at posttest than comparison females. Consistency of contraceptive use increased over time among males in the program and decreased among comparison males. Delivery of group services was correlated with delayed onset of intercourse among males, and the receipt of services related to psychosocial skills was correlated with greater contraceptive use at last sex among all sexually experienced youth. CONCLUSIONS: This new program, which serves a population known to be at very high risk for early pregnancy, appears to be effective at reducing rates of pregnancy and improving several pregnancy-related risk behaviors. PMID- 12729137 TI - Evaluation of the efficacy of a nonlatex condom: results from a randomized, controlled clinical trial. AB - CONTEXT: To reduce unintended pregnancy and HIV infection, it is critical to develop reliable male condoms that will attract consumers who reject conventional latex condoms. METHODS: In a prospective clinical trial conducted in 1998-2000, 830 monogamous couples were randomized in equal numbers to use either a nonlatex condom or a commercial natural latex condom for six months as their only method of birth control. Couples completed detailed reports for the first five condom uses and recorded intercourse and condom use in coital diaries. Pregnancy rates associated with typical and consistent condom use were calculated using life table analysis. Rates of clinical failure (condom breakage or slippage) were determined for the first five condom uses. RESULTS: During the first five uses, the nonlatex condom had a higher frequency of breakage or slippage during intercourse or withdrawal (4.0%) than latex condoms (1.3%); the breakage rate for the nonlatex condom was about eight times that of latex condoms. The six-cycle typical-use pregnancy rate did not differ significantly between users of nonlatex (10.8%) and latex condoms (6.4%). The six-cycle consistent-use pregnancy rate was higher for nonlatex condom users than for latex condom users (4.9% vs. 1.0%). CONCLUSIONS: The data present strong indirect support for public health messages that promote the use of latex condoms and, for individuals who cannot or are unwilling to use latex condoms, the use of nonlatex condoms for prevention of pregnancy and disease. PMID- 12729138 TI - Preventing sexual risk behaviors and pregnancy among teenagers: linking research and programs. PMID- 12729139 TI - The measurement and meaning of unintended pregnancy. PMID- 12729140 TI - Learning from less common disorders. PMID- 12729141 TI - A population survey of mental health problems in children with epilepsy. AB - The 1999 British Child and Adolescent Mental Health Survey, a nationwide epidemiological study of rates of psychiatric disorder in children aged 5 to 15 years, provided the opportunity to investigate the mental health of children with epilepsy. These children and their families experience disability specifically because of additional emotional, behavioural, and relationship problems, and this is the first epidemiological study that directly measures these impairments. Information was obtained by interviewing a main carer and teacher for 10,316 children; 67 children with epilepsy were identified (35 males, 32 females; mean age 10 years 2 months, SD 2 years 11 months, range 5 to 15 years), and compared with the 47 children with diabetes (27 females, 20 males; mean age 10 years 4 months, SD 3 years 4 months, range 5 to 15 years) and 10,202 controls (50% male; mean age 9 years 11 months, SD 3 years 1 month, range 5 to 15 years). DSM-IV psychiatric diagnoses were derived from the Development and Well-Being Assessment in combination with the interview and a specialist clinician rating. Parental reports of emotional and behavioural problems, their impact, and associated peer problems were also obtained. Rates of psychiatric disorder were 37% (95% confidence interval [CI] 22 to 49) in epilepsy, 11% (95% CI 2 to 19%) in diabetes, and 9% (95% CI 9 to 10%) in control children. Parents of children with epilepsy consistently reported more problems, with greater impact and associated peer problems. Epilepsy, but not diabetes, was independently (adjusted for age, sex, and severe learning difficulties) associated with all behavioural variables in regression analyses. Emotional, behavioural, and relationship difficulties are common in children with epilepsy, and constitute a significant burden to the children and their families, indicating the need for effective mental health services for these children. PMID- 12729142 TI - Fatigue in adults with cerebral palsy in Norway compared with the general population. AB - Fatigue is commonly reported among adults with cerebral palsy (CP) and in the general population. The aim of this study was to examine the prevalence of perceived fatigue in adults with CP compared with the general Norwegian population. A multidimensional questionnaire, containing among other instruments the Fatigue Questionnaire, was sent to persons with CP aged 18 years or older with no intellectual disabilities documented in their medical records. Results were compared with recently published results of a study of fatigue in the general population in Norway. There were 406 respondents (209 males, 197 females; mean age 34 years, SD 11 years; age range 18 to 72 years). Distribution of the different types of CP coincided with international epidemiological studies on CP. Adults with CP reported significantly more physical, but not more mental fatigue, than the general population. The strongest predictors associated with fatigue were bodily pain, deterioration of functional skills, limitations in emotional and physical role function, and low life satisfaction. Fatigue was not significantly associated with type of CP but was most prevalent in persons with a self-reported moderate grade of CP. Results of the present study indicate that physical fatigue is a pronounced problem in adults with CP, which has an impact on the preservation of functional skills and life satisfaction. Fatigue and factors related to fatigue should be addressed specifically in follow-up programmes for persons with CP. PMID- 12729143 TI - Transient, idiopathic nystagmus in infants. AB - The aim of this study was to characterize children with transient nystagmus. Eleven children (six males, five females) developed nystagmus in infancy and then experienced regression of the problem, usually within a few months. Mean age at onset was 2.7 months, and mean age at regression was 8.5 months. No etiology could be ascertained in any of the patients, although four children had other eye or vision abnormalities (regressed retinopathy of prematurity, n=1; asymmetric fundus colobomata, n=1; delayed visual maturation, n=2). Results of this study suggest that mechanisms which allow ocular motor stability undergo a period of postnatal maturation, during which nystagmus can occur, but also during which nystagmus may disappear. Not every case of transient nystagmus should be categorized as spasmus nutans. There is a subset of infants and young children who develop transient nystagmus with no other findings and in whom the nystagmus disappears. PMID- 12729144 TI - Childhood medulloblastoma in northwest England 1954 to 1997: incidence and survival. AB - The incidence, survival patterns, and presenting symptoms of children with medulloblastoma were studied. Data were ascertained from the Manchester Tumour Registry which is population-based and has collected data on all childhood malignancies in northwest England since 1954. Incidence rates standardized to the European standard population were calculated and Poisson regression models were used to examine temporal changes in the incidence rates during the period 1954 to 1997. Kaplan-Meier survival curves were derived and used to study changes in survival patterns. World-standardized incidence rates were 5.5 per million child years in males and 3.4 per million child years in females. Incidence rates increased from the 1950s to the 1980s but have declined recently. The 5-year survival rate has improved from 29 to 58% with similar rates for males and females. The 1-year survival rate has also improved, but females had worse survival at this point (58%) than males (75%). The type of symptom or sign at presentation is strongly affected by age, with 10 of the 22 recorded symptoms or signs showing significant age differences. The older the child is, the more likely is the presentation to show pressure features of headache, vomiting, and ophthalmic signs. Younger children present with non-specific features such as lethargy, behavioural disturbance, or increasing head size. Ataxia is seen in about 75% of children across the age range. PMID- 12729145 TI - Prevalence of tic disorders and Tourette syndrome in a Swedish school population. AB - The aim of the study was to find the epidemiological distribution of tic disorders and Tourette syndrome (TS) in Swedish school children aged 7 to 15 years. A total population of 4,479 children and their parents were asked to fill in a questionnaire covering both motor and vocal tics. A three-stage procedure was used: screening, interview, and clinical investigation. Two hundred and ninety-seven children (190 males, 107 females) were found to have tics. TS, according to DSM-IV criteria, was found in 0.6% of the total population, another 0.8% had chronic motor tics, and 0.5% had chronic vocal tics. Further, 4.8% of the children had transient tics. All together 6.6% of 7- to 15-year-old children currently had or had experienced some kind tic disorder during the last year. Prevalence of different tic disorders was higher among younger children and in males, and was highly associated with school dysfunction. The prevalence of TS was higher than was previously thought but other tic disorders were more common in this childhood population. PMID- 12729146 TI - Corpus callosum and posterior fossa development in monozygotic females: a morphometric MRI study of Turner syndrome. AB - Previous neuroimaging research in Turner syndrome (TS) has indicated parietal lobe anomalies, while anomalies in other brain loci have been less well substantiated. This study focused on potential cerebellar abnormalities and possible disruptions of interhemispheric (parietal) callosal connections in individuals with TS. Twenty-seven female children and adolescents with TS (mean age 13 years, SD 4 years 2 months) and 27 age-matched female control individuals (mean age 13 years 2 months, SD 4 years 1 month) underwent MRI. Age range of all participants was 7 to 20 years. Morphometric analyses of midline brain structures were conducted using standardized, reliable methods. When compared with control participants, females with TS showed reduced areas of the genu of the corpus callosum, the pons, and vermis lobules VI-VII, and an increased area of the fourth ventricle. No group difference in intracranial area measurements was observed. The reduced area of the genu in TS may reflect compromised connectivity between inferior parietal regions. Further, cerebellar vermis hypoplasia associated with TS agrees with literature that suggests the posterior fossa as a region prone to structural alterations in the face of early developmental insult. PMID- 12729147 TI - Findings from a multidisciplinary clinical case series of females with Rett syndrome. AB - Systematic data from a multidisciplinary clinical assessment of a large series of females with Rett syndrome (RS; n=87) is presented. Participants' ages ranged from 2 years 1 month to 44 years 10 months. Areas assessed included oromotor skills, feeding problems, growth, breathing abnormalities, mobility, postural abnormalities and joint deformities, epilepsy, hand use and stereotypies, self care, and cognitive and communication skills. Many previously reported trends in the presentation of RS over time were confirmed, notably the increasingly poor growth and near pervasiveness of fixed joint deformities and scoliosis in adulthood. In contrast, there was a slight trend towards improved autonomic function in adulthood, whereas feeding difficulties increased into middle childhood and then reached a plateau. Improvements in mobility into adolescence were followed by a decline in those skills in adulthood. Levels of dependency were high, confirming findings from previous studies. Despite the presence of repetitive hand movements, a range of hand-use skills was seen in individuals of all ages. Cognitive and communication skills were limited, but there was little evidence of deterioration of these abilities with age. These findings confirm that RS is not a degenerative condition and indicate that intervention and support to maintain and increase motor skills, daily living skills, and cognitive and communicative functioning are appropriate targets for individuals with RS. PMID- 12729148 TI - Prevalence of hearing loss in Rett syndrome. AB - This study examined the auditory status of females with Rett syndrome (RS) using auditory brainstem response (ABR) testing and measures of acoustic admittance. Prevalence of hearing impairment in 81 female patients (age range 1 year 3 months to 39 years 8 months; mean age 8 years 7 months, SD 6 years 4 months) was evaluated with ABR measurements. If the ABR latencies at 20dB nHL exceeded those of the normative group, a slight hearing loss was diagnosed. For ABR thresholds in the range 30 to 40dB nHL, a mild hearing loss was determined; moderate hearing loss was diagnosed for ABR thresholds in the range 50 to 60dB nHL. Bilateral hearing loss was found in 16 (19.3%) patients and unilateral hearing loss in 13 (16%) patients. Bilateral hearing loss of moderate degree was found in one patient. No patient was found with severe hearing loss (ABR threshold of 70dB nHL and above) in either ear. Slight hearing loss was found in 10.5% of ears (17 of 162), mild hearing loss in 14.2% of ears(23 of 162), and moderate hearing loss in 3.1% of ears (5 of 162). In 72.2% of ears (117 of 162), findings indicated the presence of normal peripheral auditory sensitivity. Prevalence of sensorineural hearing loss was 17.3% (28 of 162). Evidence of conductive involvement (conductive and mixed hearing loss) was observed in 9.9% of ears (16 of 162). In this cross-sectional study, the prevalence of hearing loss was increased in older RS participants and in those with seizures requiring the use of anticonvulsants. Neither the type of hearing loss nor the presence of preserved speech seemed to be correlated with the type of mutation in methyl-CpG-binding protein 2 (MeCP2) gene that is associated with RS. PMID- 12729149 TI - Integration of suck and swallow rhythms during feeding in preterm infants with and without bronchopulmonary dysplasia. AB - The aim of this study was to define quantitative measures for assessing the integration and maturation of suck and swallow rhythms in preterm infants as they relate to each other. Fourteen preterm infants (eight males, six females; gestational age range 26 to 32 weeks) with bronchopulmonary dysplasia (BPD) and an age-matched cohort of 20 infants (10 males, 10 females; gestational age range 26 to 33 weeks) without BPD were studied weekly from time of initiation of oral feeding using simultaneous recordings of nipple and pharyngeal pressure. The integration of suck and swallow rhythms was quantified by using the coefficient of variation (COV) of the suck-swallow dyad interval. Infants without BPD had a significant correlation between increasing postmenstrual age (PMA) and decreasing COV of the dyadic interval (increasing stabilization; r=0.45; p=0.008). In the non-BPD cohort 35 weeks or less PMA, the mean dyadic COV was 0.42 (SD 0.12) versus 0.34 (SD 0.09) in those more than 35 weeks PMA (p=0.039). In contrast, dyadic stability in infants with BPD was not correlated with PMA. Infants with BPD of more than 35 weeks PMA had less dyadic stability (0.45, SD 0.10) than did age-matched controls (p<0.001). Dyadic stability was also correlated with feeding efficiency in the non-BPD group (r=0.46;p=0.007) but not in the BPD cohort. Therefore, ontogeny of rhythmic suckle feeding can be described quantitatively in preterm infants, allowing comparison with at-risk populations. Infants with BPD do not follow predicted maturational patterns of suck-swallow rhythmic integration. PMID- 12729150 TI - Balint's syndrome in a 10-year-old male. AB - A 10-year-old male was referred with difficulties at school. He had particular difficulty with reading long words, following the sequence of text down a page, writing words in the correct order, writing words in line, and copying from the blackboard. He had a history of infective endocarditis complicated by intracerebral haemorrhage at the age of three years. Detailed history taking revealed symptoms typical of 'dorsal stream' pathology, namely a deficit of 'vision for action'. This included a spatial disorder of attention (simultanagnosia), defective hand and foot movements under visual control (optic ataxia), and acquired oculomotor apraxia which are consistent with Balint's syndrome. Strategies were suggested for coping with the symptoms and one year later a distinct improvement in adapting to the disability was found. PMID- 12729151 TI - The acidosis paradox: asphyxial brain injury without coincident acidemia. PMID- 12729152 TI - 'Clinical experience of constraint induced movement therapy in adolescents with hemiplegic cerebral palsy--a day camp model'. PMID- 12729153 TI - 'Use of selective serotonin reuptake inhibitors in children with pervasive developmental disorder: risk of treatment emergent mania'. PMID- 12729154 TI - The effect of temperature on midgut and brain protein profiles in Morimus funereus larvae (Coleoptera: Cerambycidae). AB - The 7-days shift of M. funereus larvae, from nature to a constant temperature of 23 degrees C led to changes in midgut and brain protein quality and quantity. The changes in midgut protein profiles are characterized by an intensified protein band Mr of 29 kD, the absence of protein Mr of 22 kD and less intense bands Mr of 8.5-2.5 kD. Electrophoretic patterns of brain proteins showed less intense Mr of 66-2.5 kD protein bands. PMID- 12729155 TI - Transepithelial transport of sodium and chloride ions in isolated skin of the frog, Rana esculenta L. AB - Isolated frog skin, mounted in a Ussing apparatus, was investigated electrophysiologically. Application of amiloride, an inhibitor of sodium ion transport, and bumetanide, known to block the transport of chloride ions, revealed the effect of these ions on PD, both under control conditions and following mechanical stimulation. Under control conditions, mechanical stimulation of the skin caused hyperpolarization, i.e. a transient increase in the electrical potential difference. Preincubation in the presence of amiloride, or amiloride plus bumetanide, brought about both a decrease in electrical potential and an inhibition of the reaction upon stimulation. On the other hand, incubation with bumetanide resulted in a decrease in electrical potential, but did not affect the skin reaction after mechanical stimulation. The above results indicate that hyperpolarization of the frog skin following mechanical stimulation is caused by enhanced transepithelial transport of sodium ions which, in turn, is induced by stimulation of sensory receptors. PMID- 12729156 TI - Gonadotropins and sex hormones modulate interrenal function in soft-shelled turtle. AB - The aim of the current work was to investigate the role of gonadotropins and female sex hormones on interrenal activity in soft-shelled turtles, Lissemys punctata punctata. 1) FSH treatment (3 microg/100 g body wt daily for 10 days) caused interrenal hypertrophy with increased nuclear diameter, raises of acid phosphatase and alkaline phosphatase concentrations, and depletions of cholesterol (except the free fraction) and ascorbic acid levels from the interrenal gland. However, LH treatment (3 microg/100 g body wt daily for 10 days) failed to produce any perceptible change in the interrenal activity. The combined treatments of FSH and LH (3 microg each/100 gm body wt daily for 10 days) produced no further change beyond that of FSH alone. 2) Estrogen treatment with the low dose (25 microg/100 g body wt daily for 10 days) had no effect, but with higher doses (50 microg or 100 microg/100 gm body wt daily for 10 days) is caused interrenal stimulation by inducing the same manifestations to those of FSH. The degree of manifestations was higher with the higher dose (100 microg daily) than that of the moderate dose (50 microg daily). Progesterone treatment with the low dose (25 microg /100 g body wt daily for 10 days) had no significant effect, but with the moderate (50 microg daily) and higher (100 microg daily) doses suppressed interrenal activity by showing the reverse changes to those of estrogen. The degree of manifestations was higher with the higher dose than that of the moderate one. The combined treatments of estrogen and progesterone (100 microg each/100 g body wt daily for 10 days) caused interrenal stimulation but to a lesser extent than that of estrogen alone. The findings are briefly discussed. PMID- 12729157 TI - Age-dependent effect of limb immobilization and remobilization on rat bone. AB - The effect of unilateral hindlimb immobilization and subsequent free remobilization on bone tissue in rats was examined. Right hindlimb of intensively growing (G), young adult (Y) and adult (A) male rats was immobilized by taping for two weeks. Bone tissue was investigated post mortem in experimental and age matched control rats, either directly after immobilization (Imm) or after two or four weeks of remobilization (Re2, Re4). Apparent density (d(app)) and mineralization (Min) were estimated in femora and pelvis. The mechanical state of bone tissue in femora was evaluated using an ultrasonic method. Additionally, activity of serum alkaline phosphatase, and serum calcium and phosphorus were measured in each group. Min and d(app) in Imm bones were changed in G rats, while in Y and A only d(app) in Imm femora was affected. Velocity of ultrasound was significantly lower in immobilized femora in each age group, indicating decreased elasticity of bone tissue. The differences between immobilized and control limbs were still significant in Re2 and Re4 groups in G rats. In Y rats the differences between experimental and control bones increased during remobilization. It is concluded that deterioration of bone initiated during two weeks of unloading, last within at least four weeks of free remobilization, despite restoration of normal activity. PMID- 12729158 TI - Heterochromatic banding patterns on chromosomes of twelve weevil species (Insecta, Coleoptera, Curculionoidea: Apionidae, Curculionidae). AB - The C-banding patterns of twelve weevil species are presented. The obtained results confirm the existence of two groups of species: with a small or large amount of heterochromatin in the karyotype. The first group comprises seven species (Apionidae: Holotrichapion pisi; Curculionidae: Phyllobius urticae, Ph. pyri, Ph. maculicornis, Tanymecus palliatus, Larinodontes turbinatus, Cionus tuberculosus). In weevils with a small amount of heterochromatin, tiny grains on the nucleus in interphase are visible, afterwards in mitotic and meiotic prophase appearing as dark dots. The absence of C-bands does not indicate a lack of heterochromatin but heterochromatic regions are sometimes so small that the condensation is not visible during the cell cycle. The second group comprises five species (Otiorhynchus niger, O. morio, Polydrusus corruscus, Barypeithes chevrolati, Nedyus quadrimaculatus) which possess much larger heteropicnotic parts of chromosomes visible during all nuclear divisions. The species examined have paracentromeric C-bands on autosomes and the sex chromosome X, except for Otiorhynchus niger, which also has an intercalary bands on one pair of autososomes. All the species examined differ in the size of segments of constitutive heterochromatin. The y heterochromosome is dot-like and wholly euchromatic in all the studied species. PMID- 12729159 TI - A review of karyotype variation in jumping plant-lice (Psylloidea, Sternorrhyncha, Hemiptera) and checklist of chromosome numbers. AB - Karyotype variation in Psylloidea (Hemiptera) was analysed. Chromosome numbers and type of chromosome sex determination of 153 species belonging to 51 genera and five out of six families (except tropical Phacopteronidae) are listed. Some important cytological characteristics, such as chromosome type, structure of karyotypes, sex determination type, pattern of male meiosis, type of chromosome rearrangements, and B chromosome occurrence, as well as the C-heterochromatin and NOR localization, are discussed. In addition, a comparison of chromosome systems between Psylloidea and other superfamilies of Sternorrhyncha (Aphidoidea, Coccoidea, and Aleyrodoidea) was undertaken. PMID- 12729160 TI - Structure and development of ovaries in the weevil, Anthonomus pomorum (Coleoptera, Polyphaga). II. Germ cells of the trophic chamber. AB - The analysis of the germ cell cluster formation in Anthonomus pomorum (Coleoptera, Polyphaga, Curculionidae) has revealed that both linear and branched clones of cystocytes occur in the pupa stage. In the branched clones a poorly developed polyfusome is formed and cystocytes with maximally 3 intercellular bridges were found. In the linear clones the polyfusomes are absent. Further divisions of cystocytes produce exclusively linearly arranged cells. Just after metamorphosis (Imago-A stage), the process of the germ cell membrane reduction starts. Only 2 groups of cells retain cell membranes: i.e the most anteriorly localized group of cystocytes and the posteriorly located presumptive oocytes. The former cells divide mitotically during the summer. As a result an anterior posterior gradient of the syncytialization process arises in the Imago-B stage (females preparing for hibernation). In the sexually mature females (Imago-C) the trophic chamber consists of a huge syncytial area with numerous nurse cell nuclei embedded in a common cytoplasm, and posteriorly located young oocytes surrounded by prefollicular cells. In the light of recent hypothesis concerning the germ cell cluster formation and telotrophy anagenesis in Polyphaga the significance of the presented results is discussed. PMID- 12729161 TI - Development of the tongue and taste disks in Pelobates fuscus. AB - In the tadpole of Pelobates fuscus the process of tongue formation starts at the 32nd developmental stage. In more advanced stages (older than 38th) fast anterior and faucial growth of the tongue fold has been observed. This process is accompanied by the development of the gustatory organs. The dorsal surface of the tongue fold, smooth at the beginning, in older tadpoles (developmental stages 36 39th) forms protrusions in which gustatory organs of the taste disk type (TDs) develop. In the 41 st tadpole developmental stage anlages of TDs are formed by elongated cells, located more or less perpendicularly to the surface of the tongue. The diameter of the sensory area of a TD at the 45th developmental stage amounts to 94 microm, while in metamorphosed individuals it reaches 130-140 microm. At the base of a TD the presence of basal cell morphologically similar to that of Merkel cell was observed at the 42nd developmental stage of a tadpole. Fully developed afferent synaptic connections in the sensory epithelium of a TD were found starting from the 44th developmental stage. Single synaptic vesicles with an electron-dense core were observed in gustatory cells as early as at the 41 st developmental stage of the tadpole. From the observations reported here it can be inferred that in Pelobates fuscus development of both the tongue and TDs is similar to that already described in the representatives of the Rana genus. PMID- 12729162 TI - Astereological analysis of FRTL-5 cells as an effect of thyrotropin and irradiation. AB - FRTL-5 cell line is a cloned epithelial follicular cell line from Fischer rat thyroids. This cell line expresses many of the thyroid differentiated markers in vitro. Their growth and function depend on thyrotropin (TSH) as the main regulatory hormone. In this astereological analysis, the authors concentrate on FRTL-5 nuclei as the most vulnerable part of the cell. Using morphometrical variables, they wished to discover the morphologically identifiable sign of transformation of FRTL-5 cells after irradiation and to study the effect of different TSH concentrations. FRTL-5 cells were grown in a medium of 4 different concentrations of TSH (0, 0.1, 1, 10 mU/ml) and irradiated with 0 Gy, 2 Gy, and 4 Gy. The results showed that the nuclear-cytoplasmic ratio decreases after irradiation with doses of 4 Gy or if TSH was included in the medium. The nuclear maximum diameter of FRTL-5 cells increased with higher concentrations of TSH more obviously after irradiation with 4 Gy than with 2 Gy. On the basis of astereological analysis, it was concluded that different concentrations of TSH and irradiation exert an effect especially upon FRTL-5 cell nuclei. The possible transformation of FRTL-5 cells after culturing in TSH medium and after irradiation could be confirmed by injection into an animal of the Fischer strain. PMID- 12729163 TI - Further studies on the Paramecium aurelia species complex in Greece. AB - The strains collected in Athens and the city of Egina, the Island of Egina were identified as P. primaurelia. This is the first information about finding of P. primaurelia in Greece. PMID- 12729164 TI - Stress and immunity: minireview. AB - Stress, a state of threatened homeostasis, may be induced by various physical or psychological factors (stressors), including antigenic stimulation. Stressful experiences may affect both physical/psychological well being and immune functioning of humans and animals; the ongoing immune reaction may affect other physiological functions and psychological comfort. The molecular basis of these effects involves a network of multidirectional signalling and feedback regulations of neuroendocrine- and immunocyte-derived mediators. The consecutive stages of the multistep immune reactions might be either inhibited or enhanced owing to the previous and/or parallel stress experiences, depending on the kind of stressor and the animal species, strain, gender, or age. Therefore, the final results of stress-induced alteration of immune reactions are difficult to predict. The effect of a particular stressor on immune functions varies according to the previous stress experience of the individual (e.g. social confrontation, sterile saline injection) while various stressors may act in the same or in opposite ways on the same immune parameter. In general, the efficacy of immune response depends on the neuroendocrine environment on which it is superimposed. Conversely, neural and endocrine responses depend on the concurrent immune events upon which they are superimposed. It seems that the consequences of stress on the immune functioning are generally adaptive in the short run but can be damaging when stress is chronic. PMID- 12729165 TI - The ground-nesting solitary bee, Dasypoda thoracica Baer, 1853 (Hymenoptera: Apoidea: Melittidae) and its life history. AB - Diagnosis and redescription of Dasypoda thoracica Baer, 1853 are given. Its seasonal and daily activity, host plants, and foraging distance are presented. Nest architecture and mature larva, male activity, and mating behaviour are described. Distribution and Polish localities are illustrated. PMID- 12729166 TI - Taxonomic importance of the prefemoral process of the first pair of legs in males of the genus Pseudonannolene (Diplopoda, Spirostreptida). AB - The first pair of legs in males of some groups of diplopods is modified, having a large coxa and oral projecting prefemoral process; in the family Spirostreptidae (Spirostreptida) this structure has relevance on the genus level. This paper discusses the shape, presence, and absence of the prefemoral process in the genus Pseudonannolene (Pseudonannolenidae, Spirostreptida) and suggests that this structure is an important taxonomic character in this genus and can be used for distinguishing some species. PMID- 12729167 TI - The long-term effect of cadmium exposure through food on the postnatal development of the bank vole (Clethrionomys glareolus Schreber, 1780). AB - Cadmium is well known for its toxicity to the animal body. However, its effect on pregnancy and the development of young animals is still not well understood. This study examined such effects, using bank voles captured from the wild to make the results closer to those which could be expected in the natural environment. One group of animals was fed 7 microg g(-1) cadmium in the food, a second 35 microg g(-1), and a third no cadmium, as a control. The concentrations of cadmium in the whole bodies of young bank voles were determined on the 3rd, 5th, or 10th day of life. The cadmium level in the bodies of animals exposed to 35 microg g(-1) of cadmium was significantly higher than in those from either the control group or the group receiving 7 microg g(-1) of cadmium, which did not differ from each other. The cadmium level did not change with animal age in any of the study groups. Concentrations of Zn, Cu, and Fe were also determined in the whole body of young animals, as cadmium is known to disturb the metabolism of these essential metals through antagonistic activity. Both Cu and Fe levels were negatively correlated with cadmium concentrations, while a positive correlation was found between zinc and cadmium in the young animal bodies. Also found was higher offspring mortality in the group receiving 35 microg g(-1) of cadmium in food. There was no difference in young animal body weight between the study groups. PMID- 12729168 TI - The effect of heavy metal accumulation on metallothionein content in selected tissues of bank voles and yellow-necked mice caught near a steelworks and zinc smelter. AB - The effect of cadmium, zinc, and copper accumulation on metallothionein content in the selected tissues of bank voles and yellow-necked mice trapped near the Sendzimir Steelworks in Krakow and the zinc smelter in Bukowno were analysed. The Borecka Forest was chosen as a control area. The highest cadmium concentration, 32.98 microg g(-1) dry weight, was detected in the kidneys of the bank voles caught in the Bukowno area. Zinc and copper concentrations in the tissues did not exceed the critical values. Metallothionein content in the liver and kidneys was associated with heavy metal accumulation in the tissues. The highest content of sulphydryl groups was detected in the livers of the bank voles trapped within the neighbourhood of the zinc smelter in Bukowno. The highest level of disulphide bonds was found in the kidneys of the bank voles from the same area. PMID- 12729169 TI - The first report on the occurrence of twins in a freshwater crayfish, Pacifastacus leniusculus (Decapoda, Astacoidea). AB - Twin stage 1 juveniles from the signal crayfish, Pacifastacus leniusculus, were observed in this study. No such observations have been found in the literature. The twins consist of two stage 1 juveniles that are fused at the head. Each juvenile has a separate abdomen and thorax and has the appropriate number of appendages and a telson, all characteristic of a normal stage 1 juvenile. PMID- 12729170 TI - Constitutive heterochromatin in karyotypes of two Cicadidae species from Japan (Cicadoidea, Hemiptera). AB - Karyotypes of males of cicadas Tibicen bihamatus (Motschulski) and Platypleura kuroiwae Matsumura were studied using C-banding technique. In Tibicen bihamatus two types of C-band distribution were observed. Two chromosome pairs have C-bands at one of the chromosome ends, while in the other, including the sex chromosome, C-heterochromatin blocks occurred at both ends. Platypleura kuroiwae has a smaller amount of C-heterochromatin located as small subterminal blocks. The intercalar C-bands were seen in the early spermatogonial metaphase chromosomes. PMID- 12729172 TI - Douglas S Leathar Award. PMID- 12729171 TI - Paramecium tredecaurelia of the Paramecium aurelia complex in Israel. AB - The paper concerns the finding of a new habitat (Kiryat Motzkin, north of Haifa, Israel) of Paramecium tredecaurelia from the P. aurelia complex. This is only the forth known locality of the species in the world. Previously, its strains were obtained from widely separated localities: the River Seine, Paris, France; Benenitra, Madagascar, and the Cuernavaca Valley, Taxco, Mexico. The studied strain originating from Israel was identified as P. tredecaurelia on the basis of the strong (90%) conjugation between the complementary mating type of the examined clones with the appropriate standard strain 209 of P. tredecaurelia from Paris, France (restricted to odd mating type). However, the strain from Israel is restricted to the even mating type. PMID- 12729173 TI - Meaning and knowledge of health among older adult immigrants from Russia: a phenomenological study. AB - This qualitative study examined the meaning and knowledge of health among older adult Russians who have immigrated to the US. Prior studies have shown that this group has a high rate of chronic disease and low compliance with preventative health behaviors. However, little is known about the knowledge and beliefs about health among Russian immigrants that may be driving their low participation in health behaviors. The goal of the present study was to use a phenomenological approach to gain a deeper understanding of the experience of health in this population. Twelve older adults were recruited through English language classes, and interviewed in focus groups about their experience with health and health care. Qualitative analysis using the phenomenological approach revealed a number of important health themes: participants (1) define health as the absence of disease, (2) feel distrust toward media information about health, (3) experience alienation from their current health care system, and (4) experience a sense of stress and helplessness in the US because they do not understand the English language or the US health care system. These themes may underlie the immigrants' lack of participation in health practices and in seeking out information about health. Information about the Russian immigrants' knowledge about health behavior is also reported. This study represents a first step toward better understanding the barriers facing older adult Russian immigrants in learning about and practicing health behaviors. The study also draws attention to the ways in which beliefs about health may impact health behavior. PMID- 12729174 TI - Beliefs as predictors of condom use by injecting drug users in treatment. AB - This study was conducted to clarify (1) the extent to which health beliefs selected from Protection Motivation Theory can combine to correctly classify 72 injecting drug users (IDUs) as condom users or non-users and (2) which of the beliefs ('vulnerability to a regular partner', 'vulnerability to a casual partner', 'self-efficacy', 'response efficacy', 'response costs' and 'social norms') were most influential in this distinction. Results of a logistic regression indicated that these beliefs were significant predictors of condom use. Overall, 83.3% of participants were correctly classified according to condom use, with condom 'non-users' being more accurately predicted (94.0%) than 'users' (59.1%). 'Vulnerability to a regular partner' and 'social norms' were significant multivariate and univariate predictors of condom use, and 'response costs' were significant univariate predictors. IDUs were confident of their ability to use condoms, considered themselves highly vulnerable to HIV infection from casual partners and were confident in the efficacy of condoms to protect them from AIDS. However, the majority of IDUs were not condom users, particularly with 'regular' partners. Findings suggest that HIV prevention programmes should target beliefs regarding risks from known partners, perceived norms and negative consequences of condom use in order to increase condom use by IDUs in treatment. PMID- 12729176 TI - Male involvement in family planning in rural Vietnam: an application of the Transtheoretical Model. AB - Various studies have shown that family planning adoption is likely to be more effective for women when men are actively involved. Male involvement in family planning is an instance of behavior change. The Transtheoretical Model of behavior change was used to examine men's involvement in general contraception and intrauterine device (IUD) use by their wives. The study tested whether the constructs of the model, decisional balance and self-efficacy, are sensitive to differences in stages of change. Reliable scales to test decisional balance and self-efficacy were developed. The study was carried out in rural Vietnam with 201 eligible participants. The staging algorithm identified that 25.8% of men were in the precontemplation stage, 10.5% of men were in the contemplation/preparation stages and 63.7% of men were in the action/maintenance stages. Disadvantages of IUD use for men in precontemplation were significantly higher than those in the action/maintenance stages, while the reverse was true for self-efficacy for convincing their wives to use an IUD. Interventions that are targeted to stage of change, that seek to reduce cons and that increase self-efficacy have the potential to influence male involvement in IUD adoption by their wives. PMID- 12729175 TI - Understanding the potential of teachable moments: the case of smoking cessation. AB - The label 'teachable moment' (TM) has been used to describe naturally occurring health events thought to motivate individuals to spontaneously adopt risk reducing health behaviors. This manuscript summarizes the evidence of TMs for smoking cessation, and makes recommendations for conceptual and methodological refinements to improve the next generation of related research. TM studies were identified for the following event categories: office visits, notification of abnormal test results, pregnancy, hospitalization and disease diagnosis. Cessation rates associated with pregnancy, hospitalization and disease diagnosis were high (10-60 and 15-78%, respectively), whereas rates for clinic visits and abnormal test results were consistently lower (2-10 and 7-21%, respectively). Drawing from accepted conceptual models, a TM heuristic is outlined that suggests three domains underlie whether a cueing event is significant enough to be a TM for smoking cessation: the extent to which the event (1) increases perceptions of personal risk and outcome expectancies, (2) prompts strong affective or emotional responses, and (3) redefines self-concept or social role. Research in TMs could be improved by giving greater attention to assessment of conceptually grounded cognitive and emotional variables, appropriately timed assessment and intervention, and inclusion of appropriate target and comparison samples. PMID- 12729177 TI - Print material content and design: is it relevant to effectiveness? AB - Printed materials are widely used in cancer education. There are a considerable number of guidelines in the literature on the content and design characteristics of print materials. However, there is little outcome-based evidence about whether materials containing these characteristics are more effective under real-world conditions. Six pamphlets were designed such that two had most of these characteristics, one had content characteristics only, one had design characteristics only and two had few of the characteristics. The pamphlets encouraged women aged 50-69 years to join a Pap Test Reminder Service (PTRS). The pamphlets were mailed to a randomly selected sample of 7668 women. Registrations with the PTRS were monitored. Receipt, readership, acceptability, knowledge and attitudes were assessed by telephone interview. There were no significant differences between the pamphlet groups in registrations with the PTRS, receipt, readership, acceptability, knowledge or attitudes. It was concluded that the inclusion of content and design characteristics does not result in more effective print materials. PMID- 12729178 TI - Patients' perspectives on diabetes health care education. AB - Living with Type 2 diabetes requires that patients develop a range of competencies that allow them to take greater control over the treatment of their disease. This requires education that promotes health whilst respecting individuals' self-perceived needs and voluntary choices. Whilst such a concept is not new in the field of diabetes, health professionals are still struggling with how to administer it successfully. This paper presents the findings of a research trial of a theoretically constructed educational intervention. It focuses on the patients' perspectives of what they valued about the intervention which was found to be clinically effective over a short-term period only. Limitations to maintaining effects were associated with a number of factors. The study found that whilst patients can be educated toward greater autonomy, not all health professionals are ready to work in partnership with them. It highlighted the importance of clinical staff not only gaining a better understanding of diabetes management, but also of the theoretical principles underlying patient empowerment. This paper outlines these principles and shows how they were synthesized to produce a framework for informing practice. Patients' views are utilized to provide guidelines for improving the outcomes of patient education. PMID- 12729179 TI - Promoting mental health through employment and developing healthy workplaces: the potential of natural supports at work. AB - In England, policy developments in the field of mental health are stimulating interest in employment for mental health service users as a means of mental health promotion. To date, research that might assist in increasing employment rates amongst this group has focused largely on the question of which service users are most likely to benefit from vocational interventions and, more recently, on models of vocational support. Less is known about how employers can assist people in their transition or return to work. In this article we draw on the accounts of 17 employment project clients to identify workplace factors that were associated with job retention. Specific adjustments such as flexibility about working hours, work schedules and job tasks emerged as crucial in enabling clients to deal with the effects of medication, and to regain stamina and confidence. Over and above these, however, 'natural supports' of a kind from which any employee would arguably benefit were equally important. In this respect the main themes revolved around training and support to learn the job, supportive interpersonal relationships at work, workplace culture, and approaches to staff management. These themes might equally provide a productive focus for workplace health promotion more generally, using organization development approaches. PMID- 12729180 TI - Oral tobacco cessation with UK resident Bangladeshi women: a community pilot investigation. AB - Our objective was to establish the short-term outcomes for successful tobacco cessation of a programme offering UK resident Bangladeshi women chewing paan with tobacco nicotine replacement therapy (NRT) in addition to brief advice and encouragement alone. We used a short-term longitudinal, quasi-experimental study design, in the setting of two local authority housing estates in Tower Hamlets, London. Bangladeshi women volunteers were recruited following presentations to community groups. The volunteers were assigned, after matching for age, number of paan with tobacco chewed daily and medical screening, to receive one of two tobacco cessation interventions (NRT with brief encouragement and advice, and brief advice and encouragement alone). The main outcome measures were changes in tobacco use and nicotine dependence, assessed by questionnaire and intake measures, adverse effects, and withdrawal symptoms. In total, 130 volunteers were recruited. Their mean age was 42.5 years (SD = 11.3). Mean number of paan quid with tobacco chewed daily was 10.7 (SD = 9.3) and the average age of starting to add tobacco to paan was 24 years (SD = 12). Ninety-one percent completed the 4 week trial. We found that 19.5% had stopped tobacco use, of whom 22% had received NRT, and 17% brief advice and encouragement alone. The successful members of the NRT group made a significantly greater reduction in their salivary cotinine scores at final review compared to baseline. Oral pain was reported as a barrier to successful oral tobacco cessation by 62% of the volunteers at final review. We conclude that methods identified as helping tobacco smokers successfully stop smoking can be used with Bangladeshi women chewing paan with tobacco. More research is needed to investigate these short-term outcomes and to explore the particular barriers to successful cessation for this group such as oral pain. PMID- 12729181 TI - Assessing stages of change for fruit and vegetable intake in young adults: a combination of traditional staging algorithms and food-frequency questionnaires. AB - Adequate fruit and vegetable intake is strongly associated with a reduced risk for various chronic diseases. US national surveys show that 18- to-24-year-olds are not consuming enough of these foods. Theory-based nutrition interventions, e.g. stage-tailored education programs, are needed for promoting fruit and vegetable consumption in this age group. Accurate stage assignment is the basis for developing effective stage-tailored interventions. In the current study, three different methods were compared for assigning stages of change in fruit and vegetable intakes by young adults. Significant differences in food intake, decisional balance and self-efficacy were found between respondents with concordant responses to the traditional stage algorithm and the food-frequency questionnaire (FFQ) and those with discordant responses. The stage assignment method that combined the staging algorithm and FFQ identified a distinct stage, labeled 'non-reflective action', in addition to the traditional five stages of change. This stage lay between the preparation and action stages with regard to food intake and psychosocial variables. Implications of the findings were discussed for future intervention programs that attempt to promote fruit and vegetable intake. PMID- 12729183 TI - Global health promotion and health education initiatives related to chronic disease prevention. PMID- 12729182 TI - A review of research on fidelity of implementation: implications for drug abuse prevention in school settings. AB - To help inform drug abuse prevention research in school settings about the issues surrounding implementation, we conducted a review of the fidelity of implementation research literature spanning a 25-year period. Fidelity has been measured in five ways: (1) adherence, (2) dose, (3) quality of program delivery, (4) participant responsiveness and (5) program differentiation. Definitions and measures of fidelity were found not to be consistent across studies, and new definitions are proposed. While there has been limited research on fidelity of implementation in the social sciences, research in drug abuse prevention provides evidence that poor implementation is likely to result in a loss of program effectiveness. Studies indicate that most teachers do not cover everything in a curriculum, they are likely to teach less over time and training alone is not sufficient to ensure fidelity of implementation. Key elements of high fidelity include teacher training, program characteristics, teacher characteristics and organizational characteristics. The review concludes with a discussion of the tension between fidelity and reinvention/adaptation, and ways of resolving this tension. Recommendations are made for developing a consistent methodology for measuring and analyzing fidelity of implementation. Further, researchers and providers should collaborate to develop ways of introducing flexibility into prevention programs. PMID- 12729184 TI - A national outbreak of multi-resistant Salmonella enterica serovar Typhimurium definitive phage type (DT) 104 associated with consumption of lettuce. AB - Between 1 August and 15 September 2000, 361 cases of Salmonella enterica serotype Typhimurium definitive phage type (DT) 104, resistant to ampicillin, chloramphenicol, streptomycin, sulphonamides, spectinomycin and tetracycline (R type ACSSuSpT), were identified in England and Wales residents. Molecular typing of 258 isolates of S. Typhimurium DT104 R-type ACSSuSpT showed that, although isolates were indistinguishable by pulsed-field gel electrophoresis, 67% (174/258) were characterized by a particular plasmid profile. A statistically significant association between illness and consumption of lettuce away from home was demonstrated (OR = 7.28; 95% CI=2.25-23.57; P=0.0006) in an unmatched case control study. Environmental investigations revealed that a number of food outlets implicated in the outbreak had common suppliers of salad vegetables. No implicated foods were available for microbiological testing. An environmental audit of three farms that might have supplied salad vegetables to the implicated outlets did not reveal any unsafe agricultural practices. The complexity of the food supply chain and the lack of identifying markers on salad stuffs made tracking salad vegetables back to their origin extremely difficult in most instances. This has implications for public health since food hazard warnings and product withdrawal are contingent on accurate identification of the suspect product. PMID- 12729185 TI - Environmental isolates of Citrobacter braakii that agglutinate with Escherichia coli O157 antiserum but do not possess the genes responsible for the biosynthesis of O157 somatic antigen. AB - While searching for Escherichia coli O157 in the aquatic environment of Calcutta using an immunodetection procedure, we fortuitously detected five strains of Citrobacter braakii, which cross-reacted with the commercially available O157 polyvalent antiserum. The five C. braakii isolates gave positive results when a sensitive dot-ELISA was performed with E. coli O157 monoclonal antibody. Further, the O157 monoclonal antibody recognized the bands of proteinase K treated whole cells of lipopolysaccharide of all the C. braakii isolates. Apart from weak reactions with two or three of the DNA probes, all the C. braakii strains did not hybridize with the other probes spanning the minimum region required for O157 O antigen biosynthesis. These strains did not possess any of the virulence genes that are commonly found in the Shiga toxin-producing E. coli (STEC) specially the serotype O157: H7. Therefore, it appears that the serological cross-reaction between C. braakii and E. coli O157 antiserum is based on structural mimicry between the O-polysaccharide of C. braakii and E. coli O157. PMID- 12729186 TI - Outbreak of Pseudomonas aeruginosa folliculitis associated with a swimming pool inflatable. AB - On 18 February 2002, the Communicable Disease Unit was notified by the local Public Health Service Laboratory of a child with a positive skin swab for Pseudomonas aeruginosa. This child had attended the local swimming pool and played on an inflatable, subsequently presenting to a Primary Care Nurse Practitioner with folliculitis. A total of 35 cases was identified during the outbreak. This paper describes a case-control study and microbiological sampling of the cases, the suspected inflatable and a survey of 10 swimming pool inflatables in the local area. The odds ratio for developing folliculitis following use of the inflatable was 12 (95% CI 1.05-136.80). The strain of P. aeruginosa found on the inflatable was identical to that obtained from skin swabs of cases. Nine of 10 (90%) of the inflatables sampled were colonized by P. aeruginosa. Attention should be given to the problem of routine decontamination of swimming pool inflatables. P. aeruginosa folliculitis needs to be considered in the differential diagnosis of skin rashes in children, especially in Primary Care. PMID- 12729187 TI - FcgammaRIIa and FcgammaRIIIb polymorphisms were not associated with meningococcal disease in Western Norway. AB - Fcgamma-receptor (FcyR) polymorphisms have been associated with acquisition and severity of invasive meningococcal disease. We studied FcgammaR polymorphisms in a population with a high incidence of meningococcal disease. Fifty meningococcal disease patients aged 14-60 years, with bacteriologically confirmed disease and without detected complement deficiency, together with 100 healthy adult controls were included in the study. Clinical and bacteriological data were collected prior to FcgammaRIIa and FcgammaRIIb genotyping, which was performed by polymerase chain reaction. The distribution of the FcgammaRIIa and FcgammaRIIIb allotypes and their allele frequencies were not significantly different amongst the patients and the controls. The combination FcgammaRIIa-R/R and FcgammaRIIb Na2/Na2 was less common among patients than controls (OR = 0.11, Fisher's exact P = 0.05). No significant association was found between the two FcgammaRs and severity of disease, meningococcal serogroup, age groups or gender. In contrast to previous findings, our study indicates that in Norwegian teenagers and adults, the FcgammaRIIa and FcgammaRIIIb allotypes are not decisive for the acquisition or for the severity of meningococcal disease. PMID- 12729188 TI - Analysis of longitudinal bacterial carriage studies accounting for sensitivity of swabbing: an application to Neisseria meningitidis. AB - Longitudinal carriage studies of colonizing bacteria such as Neisseria meningitidis can provide important insights into the transmission dynamics of these organisms. Carriage is detected by culturing from a nasopharyngeal swab, but the sensitivity of this technique is low and varies between studies. This paper applies a statistical method for estimating the sensitivity of swabbing, infection rate, recovery rate and initial prevalence of carriage to three longitudinal carriage studies of N. meningitidis. These parameters and 95% confidence intervals were estimated using maximum likelihood techniques. The sensitivity of swabbing was estimated to be 60-83% and this should be taken into account when interpreting carriage studies. The estimates of force of infection and recovery rates seem to be consistent with estimates from more traditional methods. Differences in the parameter estimates between datasets may be due to differences in study design. This method could be used to assist in the design of future carriage studies. PMID- 12729189 TI - Coagulase gemne variants associated with distinct populations of Staphylococcus aureus. AB - An identifying characteristic of Staphylococcus aureus is the production of staphylocoagulase (coagulase). The aim of this study was to determine the clonal distribution of coagulase gene (coa) variants within populations of S. aureus defined by multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE), and protein A variation. The N-terminal region of the coa gene from 43 methicillin-susceptible (MSSA) and 252 methicillin-resistant (MRSA) S. aureus human isolates and 9 animal S. aureus isolates was amplified and digested with HinfI. Twelve types were identified amongst the MSSA isolates and the majority (93%) of MRSA isolates were assigned to 5 of the 12 types. MLST and PFGE analysis identified epidemic populations of MRSA and each epidemic population was characterized by a different coagulase type. Nine of the 12 MLST-defined clonal complex ancestral genotypes recently described each carried a different coagulase type suggesting that coagulase evolution and the evolution of the clonal complexes are intimately related. PMID- 12729190 TI - Assessment of neonatal tetanus elimination in an African setting by lot quality assurance cluster sampling (LQA-CS). AB - Neonatal tetanus (NT) elimination, < 1 case per 1,000 live births (LB), was assessed at district level in Zimbabwe using a combined lot quality assurance cluster sampling survey (LQA-CS). Three of the highest risk districts were selected. NT was considered eliminated if fewer than a specified number of NT deaths (proxy for NT cases) were found in the sample determined using operating characteristic curves and tables. TT2 + vaccine coverage was measured in mothers who gave birth 1-13 months before the survey and women aged 15-49 years. NT was considered as eliminated, TT2+ coverage was 78% (95% CI 71-82%) in women aged 15 49 and 83% (95% CI 76-89%) in mothers. The survey cost 30,000 US dollars excluding costs of consultants. NT incidence was below the elimination threshold (< 1/1,000 LB) in the surveyed districts and probably in all districts. LQA-CS is a practical, relatively cost effective field method which can be applied in an African setting to assess NT elimination status. PMID- 12729191 TI - Smoking and polymorphisms of fucosyltransferase gene Le affect success of H. pylori eradication with lansoprazole, amoxicillin, and clarithromycin. AB - Identification of factors influencing success of Helicobacter pylori (HP) eradication is important for clinical practice. We have prospectively conducted an HP eradication study in the Aichi Cancer Center with a total of 142 patients available for analysis. The overall success rate was 61.3% (95% confidence interval 52.7-69.3%). Smoking during the medication for eradication significantly decreased the success rate (42.9%), whereas smoking cessation during the treatment was associated with a similar rate as for non-smokers (66.7%). We also examined links between an eradication outcome and polymorphisms of Le, Se, IL1A, IL1B, IL1RN and MPO genes, but with one exception none showed any association. The non-functional le allele of Le polymorphisms, leading to decreased expression of Le(b) antigen to which HP attaches with adhesin, showed a beneficial effect for success. Although further clarification is necessary, our study indicated that smoking cessation and Le gene polymorphisms may affect the success rate of HP eradication. PMID- 12729192 TI - Molecular study on human tuberculosis in three geographically distinct and time delineated populations from ancient Egypt. AB - We describe the molecular identification of human tuberculosis (TB) from vertebral bone tissue samples from three different populations of ancient Egypt. The specimens were obtained from the predynastic to early dynastic necropolis of Abydos (7 individuals, c. 3500-2650 B.C.), from a Middle Kingdom to Second Intermediate Period tomb of the necropolis of Thebes-West (37. c. 2100-1550 B.C.) and from five further Theban tombs used in the New Kingdom and the Late Period (39, c. 1450-500 B.C.). A total of 18 cases tested positive for the presence of ancient DNA (aDNA) of the M. tuberculosis complex. Out of the 9 cases with typical macromorphological signs of tuberculous spondylitis, 6 were positive for mycobacterial aDNA (66.7%). Of 24 cases with non-specific pathological alterations, 5 provided a positive result (20.8%). In 50 cases of normally appearing vertebral bones 7 tested positive (14.0%). There were only minor differences in the frequencies between the three populations. These data strongly support the notion that tuberculosis was present and prevalent in ancient Egypt since very early periods of this civilization. The unexpectedly high rate of mycobacterial aDNA in normal bone samples is presumably due to a pre- to perimortal systemic spread of the bacteria and indicates a generalized infection by M. tuberculosis. PMID- 12729193 TI - First epidemic of aseptic meningitis due to echovirus type 13 among Spanish children. AB - Echoviruses are the commonest cause of aseptic meningitis (AM). Echovirus type 13 (EV-13) was the second enterovirus serotype associated with different local outbreaks of AM in Spain between February and October 2000. It was the first time that an epidemic AM caused by this virus was recognized in Spain. The index case appeared in the Canary Islands (Canarias). The EV-13 virus was isolated from 135 patients, predominantly from cerebrospinal fluid (CSF). All isolates were from children under 13 years. The age specific peak incidence was in infants under 1 year. Most patients had fever, headache and other meningeal signs. This enterovirus serotype, not previously detected in Spain, caused severe illness with a high attack rate. PMID- 12729194 TI - An outbreak of echovirus 13 meningitis in central Israel. AB - Until recently, echovirus 13 has been a very rare cause of aseptic meningitis. We investigated an outbreak of echovirus 13 in central Israel during the summer of 2000 using a prospective case control study and a retrospective study. Echovirus 13 was isolated from 79 cerebrospinal fluid (CSF) specimens from different medical centres in central Israel. Patients' ages ranged from 10 days to 41 years (95% < 15 years, M/F ratio 62/38). A total of 128 patients with clinical aseptic meningitis were admitted to the Department of Pediatrics during the outbreak (aged 10 days to 18 years, mean 5.4 years), and 58 CSF samples were processed for viral cultures. Thirty of them did not grow any virus, 26 samples yielded echovirus 13, and 2 samples echovirus 7. The clinical features of patients with echovirus 13 in the CSF were similar to those in whom no virus was isolated or those infected with other enteroviral strains except for higher rate of fever on admission, and prolonged time with fever following the diagnosis in the echovirus 13 patients. CSF cell count varied from 4 to 2,333 cells/mm3 with polymorphonuclears (PMN) predominant in 90% of our patients. In a case-control study there was no significant difference between patients and matched controls with regard to parameters such as: day care attendance, recreation in summer camp, swimming pools and at the beach, and consumption of tap water. All the patients in our series recovered fully with no neurological abnormalities. The illness caused by echovirus 13 was benign and involved mainly patients younger than 15 year of age. Several features that characterized this outbreak include relatively high WBC in the blood and a minent CSF PMN response. PMID- 12729195 TI - A large outbreak of influenza A and B on a cruise ship causing widespread morbidity. AB - In September 2000 an outbreak of influenza-like illness was reported on a cruise ship sailing between Sydney and Noumea with over 1,100 passengers and 400 crew on board. Laboratory testing of passengers and crew indicated that both influenza A and B had been circulating on the ship. The cruise coincided with the peak influenza period in Sydney. Morbidity was high with 40 passengers hospitalized, two of whom died. A questionnaire was sent to passengers 3 weeks after the cruise and 836 of 1,119 (75%) responded. A total of 310 passengers (37%) reported suffering from an influenza-like illness (defined as cough, fever, myalgia and weakness) and 528 (63%) had seen a doctor for illness related to the cruise. One third of passengers reported receipt of influenza vaccination in 2000; however neither their rates of influenza-like illness nor hospitalization were significantly different from those in unvaccinated passengers. A case-control study also found no significant protective effect of influenza vaccination. With the increasing popularity of cruise vacations, such outbreaks are likely to affect increasing numbers of people. Whilst influenza vaccination of passengers and crew may afford some protection, uptake and effectiveness may not be sufficient to prevent outbreaks. Surveillance systems and early intervention measures, such as antiviral therapies, should be considered to detect and control such outbreaks. PMID- 12729196 TI - Economic evaluation of options for measles vaccination strategy in a hypothetical Western European country. AB - In this study an analysis was made of economic costs and medical effects (by cost effectiveness and cost-benefit analysis) associated with measles vaccination in a hypothetical Western European country. We analysed ten vaccination options in terms of past and future vaccination coverage. We show that several of the proposed strategies for improving measles vaccination coverage are preferable to maintaining the existing policies, regardless of past coverage and the viewpoint of the analysis. For society, very high coverage (95%) two-dose vaccination is most optimal, irrespective of past vaccination coverage. The addition of a one time campaign (to reduce susceptibility in (pre-)adolescent age groups) to such a high coverage two-dose vaccination programme is cost-saving to the health-care payer and to society when coverage in the past was low (< or = 70%). Even when coverage in the past was high (90%) for more than a decade, this 'maximum strategy' could be implemented at an acceptable cost to the health-care payer (incremental direct costs per discounted life-year gained < 30,000 Euros), and at net savings to society. PMID- 12729197 TI - Improving sensitivity of oral fluid testing in IgG prevalence studies: application of mixture models to a rubella antibody survey. AB - A method for the analysis of age-stratified antibody prevalence surveys is applied to a previously reported survey of antibody to rubella virus using oral fluid samples in which the sensitivity of the assay used was shown to be compromised. The age-specific distribution of the quantitative results of antibody tests using oral fluids is modelled as a mixture of strong positive, weak positive and negative components. This yields maximum likelihood estimates of the prevalence at each age and demonstrates that, when used in conjunction with mixture modelling techniques, the results of antibody prevalence studies using oral fluids accurately reflect those obtained using sera. PMID- 12729198 TI - Injections in health care settings: a risk factor for acute hepatitis B virus infection in Karachi, Pakistan. AB - A case control study was conducted to identify the association of therapeutic injections with acute hepatitis B virus (HBV) infection in Karachi, Pakistan. We enrolled 67 cases of acute HBV infection (IgM anti-HBc positive) and 247 controls (anti-HBc negative) from four hospitals of Karachi during July 2000-June 2001. Exposure to various risk factors during the period relevant to the incubation period of HBV was recorded both from cases and controls using a structured questionnaire. Multivariate logistic regression analysis of the data showed that cases were more likely to have received one injection (OR = 4.0; 95 % CI 1.4, 11.1), or more than one injection (OR = 6.3; 95 % CI 3.2, 12.4) compared to controls. The estimated population attributable risk (PAR) for therapeutic injections was 53%. Also the cases compared to controls were more likely to have household size of seven or more (OR = 1.9; 95 % CI 0.95, 3.9). This study showed that unsafe therapeutic injections appear to be the major risk factor for acute HBV infection and needs immediate focus from public health stand point. PMID- 12729199 TI - Factors influencing the shedding of verocytotoxin-producing Escherichia coli O157 by beef suckler cows. AB - A study was designed to investigate management factors that might influence the shedding of verocytotoxin-producing Escherichia coli (VTEC) O157 by beef cows in Scotland, where there is a particularly high rate of human infection. Thirty-two herds were visited at least monthly over approximately 1 year for collection of fresh faecal pat samples and information on management factors. The faecal pat samples were tested for VTEC O157 by established culture and immunomagnetic separation methods. Questionnaires were completed at the monthly visits to record management factors. Data were analysed using both univariate and multi-factor (GLMM) analysis. Changes in the number of cows in a group, dogs, wild geese, housing, and the feeding of draff (distillers' grains) were statistically significant as risk factors. The event of calving appeared to reduce the likelihood of shedding. Any effects of weaning or turnout were not statistically significant. It appears that the rate of shedding of VTEC O157 is influenced by several factors but possibly the most important of these are the circumstances of animals being housed, or, when outside, the presence of wild geese. PMID- 12729200 TI - Prevalence and characterization of Vero cytotoxin-producing Escherichia coli isolated from diarrhoeic and healthy sheep and goats. AB - Faecal samples from 146 diarrhoeic lambs and goat kids, and from 511 healthy sheep and goats were screened for the presence of Vero cytotoxin-producing Escherichia coli (VTEC). In healthy sheep and goats, VTEC were isolated in 24.4 and 16.2% of the animals, respectively. Moreover, VTEC were detected in 3.1 and 5.9% of the diarrhoeic lambs and goat kids, respectively. These data suggest that VTEC seems not to be associated with diarrhoea in lambs and goat kids. Only four VTEC strains were eae-positive. The absence of the eae gene in most of these VTEC strains could indicate that these strains are less virulent for humans that the classical eae-positive enterohaemorrhagic E. coli types. However, almost half (42.9%) and 12.2% of VTEC strains isolated from healthy sheep and goats, respectively, belonged to serotypes associated with severe diseases in humans. PMID- 12729201 TI - A one-year study of campylobacter carriage by individual Danish broiler chickens as the basis for selection of Campylobacter spp. strains for a chicken infection model. AB - From February 1999 to February 2000, 1,250 individual broiler chickens representing 125 broiler flocks originating from 62 broiler farms in Denmark were screened for campylobacter carriage. Every month, 10 flocks were tested for campylobacter carriage. The swabs were tested individually and as a pooled sample representing the flocks. Campylobacter spp. carriage was detected from 512 (40.9%) broiler chickens originating from 63 (50.4%) positive flocks. Campylobacter carriage by both individual chickens and flocks showed seasonal variation. Campylobacter jejuni was the dominant species (95.5%). Campylobacter isolates were typed using Penner heat-stable serotyping and flaA-typing methods. Data of campylobacter carriage by individual chickens and data generated by the use of different typing methods contributed to a better understanding of the dynamics of campylobacter infection within the broiler flocks. C. jejuni Penner heat-stable serotype HS2, flaA-type 1 was the most common type found in Danish broiler chickens. PMID- 12729202 TI - Clinical, epidemiological and molecular characteristics of Streptococcus uberis infections in dairy herds. AB - A longitudinal observational study (18 months) was carried out in two Dutch dairy herds to explore clinical, epidemiological and molecular characteristics of Streptococcus uberis mastitis. Infections (n = 84) were detected in 70 quarters of 46 cows. Bacterial isolates were characterized at strain level by random amplified polymorphic DNA (RAPD) fingerprinting. Persistent infections were usually attributable to one strain, while recurrent infections could be caused by different strains. When multiple quarters of a cow were infected, infections were mostly caused by one strain. In each herd, multiple strains were identified yet one strain predominated. The majority of all infections were subclinical, and infections attributed to predominant strains were more chronic than infections attributed to other strains. Epidemiological and molecular data suggest infection from environmental sources with a variety of S. uberis strains as well as within cow and between-cow transmission of a limited number of S. uberis strains, with possible transfer of bacteria via the milking machine. PMID- 12729203 TI - Time and space issues in ecotoxicology: population models, landscape pattern analysis, and long-range environmental chemistry. AB - Advances in ecotoxicology addressing problems of time and spatial scales are presented and interpreted in the frame of concepts on population/community dynamics and landscape pattern analysis. Example deterministic/probabilistic modeling experiments are used to illustrate key concepts. Space and time scales analyzed are single and multigenerations of local populations, metapopulations, community, and ecosystem/landscape. Most population models used in recent ecotoxicology studies are deterministic and do not include a formal treatment of spatial processes, like migration or local random extinction. Some metapopulation models have been applied with success. Upscaling of ecotoxicological results at the community level is less developed, probably because of the inherent complexity of indirect and direct coactions among organisms. Community and ecosystem toxicity end points that could find a broad use in regulatory applications have not yet been identified. Some practical issues like the estimation of the potential for the natural attenuation of toxicity and the transport of contaminants along food chains must be addressed at these scales/levels of biological complexity. The estimation of ecotoxicological effects has been increasingly evolving to integrate modeling and monitoring contaminant transport and fate, landscape pattern analysis, and spatially explicit population dynamics (including direct and indirect communal interactions). PMID- 12729204 TI - Ecotoxicology and spatial modeling in population dynamics: an illustration with brown trout. AB - We developed a multiregion matrix population model to explore how the demography of a hypothetical brown trout population living in a river network varies in response to different spatial scenarios of cadmium contamination. Age structure, spatial distribution, and demographic and migration processes are taken into account in the model. Chronic or acute cadmium concentrations affect the demographic parameters at the scale of the river range. The outputs of the model constitute population-level end points (the asymptotic population growth rate, the stable age structure, and the asymptotic spatial distribution) that allow comparing the different spatial scenarios of contamination regarding the demographic response at the scale of the whole river network. An analysis of the sensitivity of these end points to lower order parameters enables us to link the local effects of cadmium to the global demographic behavior of the brown trout population. Such a link is of broad interest in the point of view of ecotoxicological management. PMID- 12729205 TI - Closing the loop: a spatial analysis to link observed environmental damage to predicted heavy metal emissions. AB - In many cases, the link between industrial emissions and damage to the environment can only be inferred. The Environment Agency of the United Kingdom imposes emissions limits on industrial sites so that predicted concentrations and deposition rates remain below standard thresholds. Estimates of appropriate critical levels and loads are usually based on laboratory results and rarely estimate synergistic effects between pollutants or consider biological adaptation or selection in the target receptor organisms. The Avonmouth smelter has been emitting zinc and other heavy metals since 1929. It has been the subject of a number of detailed and synoptic studies, especially the impact on soil invertebrates. Damage was assessed using both physiological and ecological measurements. Two methods of spatial analysis were investigated, namely interpolation using standard geographical information system (GIS) operators and atmospheric dispersal modeling using an off-the-shelf model. Both methods can be used to compute contours (isolines) of predicted biological effect. Correlation results show that dispersal modeling is at least as good as kriging but requires much less data. This article demonstrates the usefulness of GIS and dispersal models as tools in decision making to determine the most suitable sampling sites in the assessment and monitoring of the impact of contamination around major point sources. PMID- 12729206 TI - Dos and don'ts of spatially explicit ecological risk assessments. AB - Location affects exposure and response to stressors at scales ranging from small sites to large regions. Tools such as geographic information systems (GIS) make it feasible to conduct spatially explicit ecological risk assessments (ERAs). However, no tools provide a panacea, and complex models based on sparse data can be inappropriate and misleading. Operations such as an interpolation within a GIS are models, and so they contain assumptions and uncertainties. Errors can propagate easily as numerous operations are performed, and the resulting uncertainties should be made explicit. Analysts should assure that space actually matters by using explanatory data prior to investigating spatial correlation. Exposure and risk estimates from experimental plots or monitoring stations may require scaling factors if they are to be used for larger watersheds or regional analyses. Maps are useful because they present complex spatial information in a manner that is easily interpreted. However, they must be prepared and interpreted with caution because they can suggest that there is a great deal more information than actually exists. In addition to presenting maps of predicted risks, maps of the spatial patterns in the uncertainties in these risk estimates should be included in risk assessments. PMID- 12729207 TI - Database mining with adaptive fuzzy partition: application to the prediction of pesticide toxicity on rats. AB - A data set of 235 pesticide compounds, divided into three classes according to their toxicity toward rats, was analyzed by a fuzzy logic procedure called adaptive fuzzy partition (AFP). This method allows the establishment of molecular descriptor/chemical activity relationships by dynamically dividing the descriptor space into a set of fuzzily partitioned subspaces. A set of 153 molecular descriptors was analyzed, including topological, physicochemical, quantum mechanical, constitutional, and electronic parameters, and the most relevant descriptors were selected with the help of a procedure combining genetic algorithm concepts and a stepwise method. The ability of this AFP model to classify the three toxicity classes was validated after dividing the data set compounds into training and test sets, including 165 and 70 molecules, respectively. The experimental class was correctly predicted for 76% of the test set compounds. Furthermore, the most toxic class, particularly important for real applications of the toxicity models, was correctly predicted in 86% of cases. Finally, a comparison between the results obtained by AFP and those obtained by other classic classification techniques showed that AFP improved the predictive power of the proposed models. PMID- 12729208 TI - Quantitative measurement of fathead minnow vitellogenin mRNA using hybridization protection assays. AB - We have developed a novel test system for the quantitative assessment of gene transcription. The procedure involves the use of chemiluminescent-labeled oligonucleotide probes in a hybridization protection assay (HPA) format. We have used this technology to measure changes in vitellogenin mRNA to demonstrate the impact of estrogen exposure in the juvenile fathead minnow (Pimephales promelas). Marked changes in mRNA expression were observed in response to intraperitoneal injection of 17beta-estradiol demonstrating the utility of this technique for the identification and monitoring of toxic responses to xenobiotics. PMID- 12729209 TI - Development of a short-term reproductive endocrine bioassay using steroid hormone and vitellogenin end points in the estuarine mummichog (Fundulus heteroclitus). AB - We have developed a short-term gonadal recrudescence test with the estuarine mummichog (Fundulus heteroclitus) and determined endocrine end points sensitive to a strong estrogen agonist (ethynylestradiol; EE2) and an antiestrogen (ZM 189,154; ZM) at concentrations of 0 to 1,000 ng/L in three separate experiments. A protocol was developed to ensure a year-round supply of recrudescing fish. A protocol for determining steroid production (testosterone and 11-ketotestosterone [11-KT] in incubated testes tissue and testosterone and 17-estradiol [E2] in incubated prematurational follicles) was optimized. Recrudescing fish (males, gonadosomatic index = 2%; females = 10%) were exposed to graded doses of EE2 or ZM for 7 to 15 d using a static daily-renewal protocol. At high EE2 (>250 ng/L), the effect on males was depression of androgen steroidogenesis and plasma steroid levels. In females, high EE2 depressed gonadal production and circulating E2 levels; however, EE2 concentrations <100 ng/L caused increased gonadal production and plasma E2. Low ZM (<100 ng/L) had little effect on male and female fish, while higher concentrations (>250 ng/L) increased E2 and 11-KT production while decreasing plasma 11-KT and E2 (1,000 ng/L only). Male and female plasma vitellogenin responded in a concentration-dependent fashion to EE2 with no effect by ZM. The low observable effect concentrations for the endocrine parameters were 1 ng/L for EE2 and 250 ng/L for ZM. The bioassay and results encompassing the environmentally relevant exposure range (1-100 ng/L) will be useful for assessing effects of endocrine-active contaminants in estuarine environments. PMID- 12729210 TI - Polycyclic aromatic hydrocarbons and black carbon in sediments of a remote alpine lake (Lake Planina, northwest Slovenia). AB - Polycyclic aromatic hydrocarbons (PAH) and black carbon (BC) were measured in alpine Lake Planina (Slovenia) sediment. Lake Planina is a remote mountain lake with almost no direct anthropogenic influence. Long-distance atmospheric deposition is a major pathway for the loading of contaminants to the sediment. The PAH were analyzed by gas chromatography coupled to mass spectrometry, whereas the BC was determined by thermal oxidation method. A flux of PAH to surface sediments of approximately 1,100 microg m(-2) year(-1) was obtained and was higher than that in other alpine lakes of the central European Alps. However, surface sediment PAH concentration, normalized to organic carbon content (OC), amounted to 5 microg PAH(pyr)/g OC and showed that Lake Planina is relatively equally exposed to atmospheric pollution compared with other lakes in the region. The BC:OC ratios ranged from 3 to 8% (w/w). In addition, a huge forest fire occurred in 1948 in the lake's surrounding area, which is recorded in the sediment. Both PAH and BC distributions were affected by the fire in 1948 in the lake's watershed, because their concentration increased remarkably. The concentration of retene, a molecular marker of coniferous wood combustion, increased to 1,000 ng/g dry weight sediment at the sediment interval corresponding to approximately the year 1950. PMID- 12729211 TI - Inverse relationship between bioconcentration factor and exposure concentration for metals: implications for hazard assessment of metals in the aquatic environment. AB - The bioconcentration factor (BCF) and bioaccumulation factor (BAF) are used as the criteria for bioaccumulation in the context of identifying and classifying substances that are hazardous to the aquatic environment. The BCF/BAF criteria, while developed as surrogates for chronic toxicity and/or biomagnification of anthropogenic organic substances, are applied to all substances including metals. This work examines the theoretical and experimental basis for the use of BCF/BAF in the hazard assessment of Zn, Cd, Cu, Pb, Ni, and Ag. As well, BCF/BAFs for Hg (methyl and inorganic forms) and hexachlorobenzene (HCB) were evaluated. The BCF/BAF data for Zn, Cd, Cu, Pb, Ni, and Ag were characterized by extreme variability in mean BCF/BAF values and a clear inverse relationship between BCF/BAF and aqueous exposure. The high variability persisted when even when data were limited to an exposure range where chronic toxicity would be expected. Mean BCF/BAF values for Hg were also variable, but the inverse relationship was equivocal, in contrast with HCB, which conformed to the BCF model. This study illustrates that the BCF/BAF criteria, as currently applied, are inappropriate for the hazard identification and classification of metals. Furthermore, using BCF and BAF data leads to conclusions that are inconsistent with the toxicological data, as values are highest (indicating hazard) at low exposure concentrations and are lowest (indicating no hazard) at high exposure concentrations, where impacts are likely. Bioconcentration and bioaccumulation factors do not distinguish between essential mineral nutrient, normal background metal bioaccumulation, the adaptive capabilities of animals to vary uptake and elimination within the spectrum of exposure regimes, nor the specific ability to sequester, detoxify, and store internalized metal from metal uptake that results in adverse effect. An alternative to BCF, the accumulation factor (ACF), for metals was assessed and, while providing an improvement, it did not provide a complete solution. A bioaccumulation criterion for the hazard identification of metals is required, and work directed at linking chronic toxicity and bioaccumulation may provide some solutions. PMID- 12729212 TI - Kinetics of benzene biodegradation by Pseudomonas aeruginosa: parameter estimation. AB - This study determined the model parameters describing biodegradation of benzene by conducting kinetic microcosm batch tests in both pure solution and saturated aquifer material conditions for various initial benzene (100-700 mg/L) and microbial concentrations (10(7)-10(9) colony-forming units [CFU]/ml) using Pseudomonas aeruginosa as benzene-degrading bacteria. In both tests, benzene and microbial concentrations were monitored over time in order to investigate which of two Monod kinetic equations, the Monod-with-growth or the Monod-no-growth model, was more suitable for describing benzene biodegradation and to estimate the associated model parameters. Parameter estimation was performed by fitting the numerical solution of each model obtained by the fourth-order Runge-Kutta integration to the measured data of benzene and/or microbial concentrations. For the Monod-with-growth model, the best fit of the numerical solution was significantly different than the measured benzene concentrations, especially at early times, because of the gradual increase of microbial population in the growth curve. In contrast, the solution based on the Monod-no-growth model produced reasonable agreement with the measured benzene data. The estimated parameters of maximum substrate utilization rate (kmax) and half-saturation constant (Kc) were in the range of 61 to 105 mg/L/d and about 270 mg/L, respectively, which differ significantly from values previously reported in the literature. We attribute the differences observed in our study to our experimental conditions of initial substrate and bacterial concentrations and oxygen and nutrient supply. Our results imply that an appropriate model type and reasonable values of kinetic parameters should be chosen to model the biodegradation of hydrocarbons in the subsurface environment. PMID- 12729213 TI - Dehydrochlorination of 1,1,1-trichloroethane and pentachloroethane by microbially reduced ferruginous smectite. AB - Reduction of structural Fe(III) in smectite clay minerals has been identified as a means to promote dechlorination of polychlorinated ethanes, but its environmental significance has yet to be fully assessed because Fe reduction has normally been achieved by agents uncommon in the environment (e.g., dithionite). This study reports the dehydrochlorination of pentachloroethane and 1,1,1 trichloroethane in the presence of ferruginous smectite reduced by two cultures of microorganisms, Shewanella oneidensis strain MR-1 (MR-R) and an enrichment culture from rice paddy soils (PS-R), in aqueous suspension under anoxic conditions. Microbially reduced ferruginous smectite facilitated dehydrochlorination of 1,1,1-trichloroethane to 1,1-dichloroethene with up to 60% conversion within 3 h of incubation time. In contrast, no formation of 1,1 dichloroethene was observed after incubation of 1,1,1-trichloroethane with chemically reduced ferruginous smectite for 24 h. Microbially reduced ferruginous smectite by MR-R and PS-R promoted the dehydrochlorination of pentachloroethane to tetrachloroethene by 80 and 15%, respectively, after 3 h of incubation time. The conversion of pentachloroethane to tetrachloroethene in the presence of chemically reduced ferruginous smectite after 24 h was 65%. These results indicate that structural Fe(II) in clay minerals has the potential to be an important reductant controlling the fate of organic chemicals in contaminated sediments. PMID- 12729214 TI - Slow-stirring method for determining the n-octanol/water partition coefficient (Pow) for highly hydrophobic chemicals: performance evaluation in a ring test. AB - The n-octanol/water partition coefficient (Pow) is one of the most important parameters employed for estimating a chemical's environmental fate and toxicity. The currently adopted test guidelines for its determination do not allow for reliable determination of log Pow greater than 5. The slow-stirring experiment, in contrast, has been demonstrated to provide reliable log Pow data up to log Pow of 8.3. To validate this method and to obtain its approval as an official Organization for Economic Cooperation and Development (Paris, France) test guideline, a ring test was performed to evaluate the accuracy and precision of the slow-stirring experiment for determination of log Pow, particularly for highly hydrophobic compounds. Up to 15 volunteer laboratories tested 1,2,3,4 tetrachlorobenzene, hexachlorobenzene. 2,2',3,3',5,5',6,6'-octachlorobiphenyl, and decachlorobiphenyl (4.5 < log Pow < 8.2). The ring-test results for the respective chemicals were 4.62, 5.50, 7.39, and 8.18. The results deviated by less than 0.1 log Pow units from the reference log Pow. In addition, the relative standard deviations of log Pow were less than 2%. Using the protocol of the ring test, log Pow of p,p'-DDT was determined to be 6.24 +/- 0.05 (mean +/- standard deviation). In combination, these results indicate that the slow-stirring method is precise and accurate and, thus, allows for reliable determination of log Pow of highly hydrophobic chemicals. PMID- 12729215 TI - Ecotoxicological responses of the mayfly Baetis tricaudatus to dietary and waterborne cadmium: implications for toxicity testing. AB - Trace metals readily accumulated by stream periphyton may enter aquatic food chains through grazer ingestion. Hence, experiments were conducted to determine the ecotoxicological responses of the grazing mayfly Baetis tricaudatus to dietary cadmium. Short-term feeding experiments indicated that B. tricaudatus nymphs did not initially avoid grazing on cadmium-contaminated diatom mats. During a partial life-cycle experiment, 4 and 10 microg/g of dietary cadmium significantly inhibited grazing, whereas 10 microg/g significantly inhibited growth. Feeding inhibition was the likely mechanism that inhibited growth (i.e., through reduced energy intake). Conversely, when exposed to waterborne cadmium using lethal toxicity test procedures, B. tricaudatus nymphs were relatively tolerant (96-h median lethal concentration, 1,611 microg/L). Thus, sublethal responses to dietary exposure appeared to be more sensitive than lethal responses to waterborne exposure. Because adult mayfly fecundity is a function of nymph size at emergence, dietary cadmium exposure could increase the extinction probability within mayfly populations. The present study highlights the importance of dietary exposure routes in determining the ecotoxicological responses of an organism to a contaminant. Furthermore, the findings emphasize the advantage of evaluating a combination of ecologically relevant, lethal and sublethal endpoints in laboratory methods used to generate data for ecological risk assessment and regulation. PMID- 12729216 TI - Toxic effects of polychlorinated biphenyl bioaccumulation in sea urchins exposed to contaminated sediments. AB - The uptake patterns and toxicity of polychlorinated biphenyl (PCB) congeners in the white sea urchin, Lytechinus pictus, on exposure to contaminated sediments were investigated. First-order modeling of uptake of the 10 most abundant PCB congeners or domains (containing more than one coeluting congener) by L. pictus indicated that a 35-d exposure was insufficient to reach steady state. Bioaccumulation of PCBs in sea urchins exhibited substantial difference between field and amended sediments, suggesting that caution must be exercised in sample preparation. Some evidence was observed of dependence of measured biota-sediment accumulation factors (BSAFs) on K(ow), indicating that equilibrium partitioning of PCBs may not always be achieved between biota lipid, sediment organic carbon, and water. Survival of L. pictus was unaffected by exposure to field and amended sediments with PCB concentrations varying more than three orders of magnitude. The growth measures (diameter, wt, and gonad wt) were significantly reduced in L. pictus exposed to San Diego Bay ([SDB]; San Diego, CA, USA) sediment, whereas they were relatively unaffected after exposure to amended sediments (with much higher PCB concentrations than SDB sediment) prepared from a New Bedford Harbor (MA, USA) sediment. The toxic effects as measured by the growth rates in L. pictus were likely attributable to polycyclic aromatic hydrocarbons (PAHs), which were elevated in SDB sediment (7.3 microg/g), rather than PCBs. PMID- 12729217 TI - Creosote toxicity to photosynthesis and plant growth in aquatic microcosms. AB - To assess photosynthesis as a bioindicator of toxicity from polycyclic aromatic hydrocarbons (PAHs), the response of chlorophyll-a fluorescence to creosote exposure was compared with effects on population-level plant growth. Large, outdoor, freshwater microcosms containing Eurasian watermilfoil (Myriophyllum spicatum) received either a single application or multiple applications of liquid creosote at nominal concentrations from 0.109 to 32.7 mg L(-1). For several weeks following treatment, photosynthetic electron transport was measured using pulse amplitude-modulated chlorophyll-a fluorescence. The maximum efficiency of photosystem II electron transport (Fv/Fm) and the quantum yield of photochemistry (deltaF/F'm) were diminished in plants shortly after the addition of creosote. In microcosms that received a single treatment of creosote only, the 50% effective concentrations (EC50s), expressed as the aqueous concentration of 15 of the most abundant PAHs, were 0.28 mg L(-1) for Fv/Fm and 0.30 mg L(-1) for deltaF/F'm. Chlorophyll-a fluorescence was diminished to a greater extent in microcosms that received multiple treatments of creosote, with EC50s of 0.13 mg L(-1) for Fv/Fm and 0.10 mg L(-1) for deltaF/F'm. Plant biomass accumulation was inhibited in a concentration-dependent manner in all microcosms treated with creosote, but this inhibition occurred to a greater degree in microcosms treated with multiple creosote applications. The response of chlorophyll-a fluorescence, measured only 8 d after creosote treatment, was similar to plant growth over the entire growing season, indicating that this technique can be used to estimate potential effects of contaminants before detrimental impacts on populations. PMID- 12729218 TI - Bioconcentration and acute toxicity of polycyclic musks in two benthic organisms (Chironomus riparius and Lumbriculus variegatus). AB - In the current study, the bioconcentration behavior and acute toxicity of two polycyclic musks, Tonalide 7-acetyl-1,1,3,4,4,6-hexamethyl-1,2,3,4 tetrahydronaphthalene (AHTN) and Galaxolide 1,3,4,6,7,8-hexahydro-4,6,6,7,8,8 hexa-methylcyclopenta[gamma]-2-benzopyran (HHCB), were studied in two benthic organisms. Polycyclic musks are frequently used fragrances, and they have been detected in different compartments of the environment. The aim of this study was to fill some empirical data gaps for AHTN and HHCB for benthic organisms. Results show that differences exist between both organisms. Chironomus riparius exhibited bioconcentration factors (BCFs) for AHTN and HHCB substantially lower than predicted for nontransformed organics. The BCFs for both chemicals increased after coexposure of the organism to the cytochrome P450 inhibitor piperonyl butoxide. Thus, the low BCF values were the result of rapid biotransformation of AHTN and HHCB in the midge larvae. Bioconcentration kinetics indicated that both chemicals induced their own cytochrome P450-mediated metabolism. Acute toxicity of AHTN to midge larvae was reduced compared to predicted baseline toxicity and was similar for HHCB. Bioconcentration of AHTN and HHCB in the worm (Lumbriculus variegatus) is in agreement with predictions based on the octanol-water partition coefficients of these chemicals. Acute toxicity was found to be similar to predicted values for baseline toxicity. Summarizing, for AHTN and HHCB, acute toxicity and bioconcentration behavior in L. variegatus was in accordance with predicted data for nontransformed organics. In C. riparius, bioconcentration as well as toxicity were reduced. PMID- 12729219 TI - A microcosm system for the study of pollution effects in shallow, sandy, subtidal communities. AB - The introduction of pollutants into the marine environment is inevitable, so an understanding of their effects is needed. Because shallow, coastal waters are at greatest risk and many of these shorelines are sandy, a test system for the shallow, sandy, subtidal environment is required. We developed a microcosm system for this purpose and tested it for three months to determine whether the benthic community remained healthy. Furthermore, because long-term effects on the environment occur primarily in the sediment, an experimental method for administering pollutants to the sediment is necessary. We administered a 1-mm layer of clean sand (simulating a technique of administering pollutants as a coating on sand) and tested the benthic community for artifactual effects of the sand itself. Fifteen metrics of community health, including the abundance of benthic microalgae, nematodes, and copepods, were measured. Most metrics were not significantly affected either by containment in the microcosms or by the addition of a 1-mm layer of clean sand. These microcosms are suitable as a test system for the shallow, sandy, subtidal environment. PMID- 12729220 TI - Availability of metals to the nematode Caenorhabditis elegans: toxicity based on total concentrations in soil and extracted fractions. AB - Current regulation of metals in soils is based on total metal concentrations rather than on actual exposure concentrations. Considering the extreme variation in soil physicochemical properties, total concentrations are not reflective of the availability and resultant toxicity of metals in different soils. In this study, the availability of Cd, Cu, Ni, Pb, and Zn to the free-living soil nematode Caenorhabditis elegans was assessed after 24-h exposures in three soils using a sequential soil extraction procedure. Albany soil, sampled from southern Georgia, USA, is characterized by a high sand content, whereas Cecil soil from the Piedmont region of Georgia contains higher fractions of clay and organic matter. The final soil was an American Society for Testing and Materials (ASTM) artificial medium composed of peat, kaolin clay, sand, and calcium carbonate. Based on their composition, ASTM medium would sorb metals most strongly and Albany soil the least strongly. In fact, 24-h lethal concentrations to 50% (LC50s) of nematodes for the five metals as determined by the total metal concentration followed this trend. In addition, water-extractable metals were lowest in ASTM medium and highest in Albany soil when spiked at the same concentrations. Our data show the need to consider soil type when performing toxicological tests and establishing site-specific allowable metal concentrations in soil. PMID- 12729221 TI - 1-Hydroxypyrene glucuronide as the major aqueous pyrene metabolite in tissue and gut fluid from the marine deposit-feeding polychaete Nereis diversicolor. AB - Both 1-hydroxypyrene and 1-hydroxypyrene glucuronide are identified as the primary phase I and phase II metabolites of the four-ringed polycyclic aromatic hydrocarbon (PAH) pyrene in the marine deposit-feeding polychaete Nereis diversicolor. Identification of pyrene and primary metabolites was performed using high-pressure liquid chromatography (HPLC) with diode-array detection and fluorescence detection (HPLC/DAD/F) and an ion-trap mass spectrometer for positive identification of 1-hydroxypyrene glucuronide. Besides 1-hydroxypyrene and 1-hydroxypyrene glucuronide, the HPLC/F trace of tissue samples from pyrene exposed worms showed three additional low-intensity peaks that may be related to pyrene metabolism based on similar excitation/emission wavelengths. The peaks were all too low in intensity to be positively identified. Of the total PAH in tissue, 1-hydroxypyrene glucuronide, 1-hydroxypyrene, and pyrene constituted 73%, 2%, and 25% respectively. Gut elimination of metabolic products is supported by the identification of 1-hydroxypyrene and 1-hydroxypyrene glucuronide in both gut fluid and defecation water. Being the only phase I metabolite of pyrene, 1 hydroxypyrene becomes a useful marker for PAH exposure, and it may serve as a valuable model compound for assessing species-specific PAH metabolic capabilities. PMID- 12729222 TI - Effects of ammonium perchlorate on the reproductive performance and thyroid follicle histology of zebrafish. AB - Adult zebrafish were reared up to eight weeks in control water or in water containing ammonium perchlorate (AP) at measured perchlorate concentrations of 18 (environmentally relevant, high) and 677 ppm. Groups of eight females were paired with four males on a weekly basis to assess AP effects on spawned egg volume, an index of reproductive performance. All treatments were applied to four to five spawning replicates. At 677 ppm, spawn volume was reduced within one week and became negligible after four weeks. At 18 ppm, spawn volume was unaffected even after eight weeks. Also, perchlorate at 18 ppm did not affect percentage egg fertilization. Fish were collected at the end of the exposures (677 ppm, four weeks; control and 18 ppm, eight weeks) for whole-body perchlorate content and thyroid histopathological analysis. Fish perchlorate levels were about one hundredth of those of treatment water levels, indicating that waterborne perchlorate does not accumulate in whole fish. At 677 ppm for four weeks, perchlorate caused thyroid follicle cell (nuclear) hypertrophy and angiogenesis, whereas at 18 ppm for eight weeks, its effects were more pronounced and included hypertrophy, angiogenesis, hyperplasia, and colloid depletion. In conclusion, an eight-week exposure of adult zebrafish to 18 ppm perchlorate (high environmentally relevant concentrations) affected the histological condition of their thyroid follicles but not their reproductive performance. The effect of 677 ppm perchlorate on reproduction may be due to extrathyroidal toxicity. Further research is needed to determine if AP at lower environmentally relevant concentrations also affects the thyroid follicles of zebrafish. PMID- 12729223 TI - Effect of cortisol treatment and/or sublethal copper exposure on copper uptake and heat shock protein levels in common carp, Cyprinus carpio. AB - This study compared the effects of increased endogenous cortisol levels and/or sublethal Cu exposure on Cu accumulation and stress protein levels (HSP70) in the freshwater common carp, Cyprinus carpio. Fish were exposed to either increased levels of endogenous cortisol (200 ng/ml) or sublethal Cu concentrations (1.9 microM, approximately 20% of the acute 96-h median lethal concentration [LC50]) alone or were pretreated by elevating plasma cortisol levels prior to Cu exposure to evaluate whether interactions between both treatments occurred. Cortisol resulted in decreased Cu levels and a decreased Cu accumulation on a short-term basis (4 h). After 96 h of Cu exposure, cortisol pretreatment resulted in augmented Cu accumulation. Exposure to Cu increased HSP70 levels in gills, erythrocytes, and liver and decreased levels in brain and kidney. No clear relationship to Cu tissue levels was observed. Increased cortisol levels or treatment with cortisol before Cu exposure decreased the HSP70 response. We can conclude that cortisol elevation results in a lower HSP70 response and thus reduces the protection against cellular stress during metal accumulation. After an initial decrease in Cu accumulation, cortisol elevation eventually stimulates metal accumulation. PMID- 12729224 TI - Effects of agricultural pesticides on the immune system of Rana pipiens and on its resistance to parasitic infection. AB - In the past 30 years, many amphibian species have suffered population declines throughout the world. Mass mortality have been frequently reported, and in several instances, infectious diseases appear to be the cause of death. The role that contaminants could play in these die-offs through immunotoxic effects has been poorly investigated. In this study, juvenile leopard frogs (Rana pipiens) were exposed for 21 d to a mixture of six pesticides (atrazine, metribuzin, aldicarb, endosulfane, lindane, and dieldrin) and subsequently challenged with a parasitic nematode, Rhabdias ranae. Exposure to the mixture at environmentally realistic concentrations significantly reduced lymphocyte proliferation. Three weeks after the end of the exposure, lymphocyte proliferation had recovered and was stimulated in frogs challenged with parasites with the exception of those previously exposed to the highest concentration. No pesticide effects on phagocytosis and splenocyte numbers were detectable at the end of the exposure period, but these two parameters were diminished 21 d after the infection challenge in frogs previously exposed to the highest levels of pesticides. In these animals, the prevalence of lung infection by R. ranae also tended to be higher. These results suggest that agricultural pesticides can alter the immune response of frogs and affect their ability to deal with parasitic infection. PMID- 12729225 TI - Exposure of small mammals, in particular the wood mouse Apodemus sylvaticus, to pesticide seed treatments. AB - Field exposure of small mammals to fungicide-treated wheat seed was investigated over three weeks following drilling on fields near York, United Kingdom. Seed consumption by small mammals trapped on and immediately adjacent to the drilled fields was quantified by measuring the amount of seed in the stomach. In addition, exposure to one seed-treatment, fluquinconazole, was quantified by measuring residues of the fungicide in the stomach, liver, and intestine. The wood mouse, Apodemus sylvaticus, was the dominant species caught on the fields and the only species found to have consumed measurable quantities of seed. Voles, Microtus agrestis and Clethrionomys glareolus, were caught in small numbers, almost exclusively in the field hedge, and showed no evidence of having consumed seed. Stomach-contents analysis revealed that more than 80% of animals trapped in the hedge adjacent to the field had consumed no wheat seed, whereas 98% had consumed less than 10% (by stomach volume). Ninety percent of animals trapped on the field had consumed seed, although 90% of these animals had less than 20% seed in the stomach. Residues of the fungicide in the stomach, intestine, and liver were lower than would be expected for the amount of seed consumed, possibly because of dehusking of the seeds by mice. The relevance of these findings when assessing exposure (and risk) posed by seed treatments to wild mammals is discussed. PMID- 12729226 TI - Breeding performance of great tits (Parus major) along a gradient of heavy metal pollution. AB - The possible effects of heavy metal pollution on various breeding parameters of the great tit (Parus major) were studied at four study sites in a pollution gradient near a large nonferrous smelter in Belgium during three consecutive breeding seasons. Significantly more females interrupted their laying period near the pollution source than did females farther away. At the two most polluted sites, hatching success was significantly reduced compared to two lesser-polluted study sites. Overall breeding success was also significantly lower toward the factory complex. This lowered breeding success was caused not only by the decreased hatching success toward the pollution source but also by the significant difference in fledging success among sites. Although a clear relationship was observed between the exposure to heavy metal pollution and an impaired reproduction, not all reproductive parameters were affected. We found no significant differences in the onset of laying, clutch size, length of the incubation period, and the proportion of nests with low-quality eggs among sites. Although nest box occupancy was every year slightly lower toward the smelter compared to farther away, differences among study sites were never significant. PMID- 12729227 TI - Bioavailability of uranium and nickel to vegetation in a contaminated riparian ecosystem. AB - The lower portion of Tims Branch (TB), a second-order stream system on the Savannah River site (SC, USA), receives influx of mixed waste-contaminated sediments from Steed Pond, a former settling basin for target processing wastes for over three decades. The magnitude and distribution of U, Ni, and other metals and the potential for trophic movement were studied to facilitate risk assessment and determine potential remedial action. Total and sequential extraction of TB soils demonstrated contaminant heterogeneity both spatially and between operationally defined fractions. Metal concentrations were elevated within riparian zone soils in contrast to stream sediments, suggesting off-site transport. Leaf tissue from TB contained an order of magnitude more Ni than tissue from reference sites. Leaves from streamside trees contained no U but elevated Ni up to 75.4 (+/-25) mg/kg dry weight (dry wt). Understory flora (Discanthelium sp. and Andropogon sp.) contained high concentrations of U associated with leaves up to 518 (+/-7.5) mg/kg dry weight U. The contrast in contaminant content and ratio of streamside and understory vegetation may result from resuspension of particulate U and Ni onto leaf surfaces and represents a potential pathway for trophic movement. The findings of this study have important ramifications for remediation of the ecosystem, suggesting that a strategy based on contaminant immobilization may be the most appropriate. PMID- 12729228 TI - Accumulation of cadmium by durum wheat roots: bases for citrate-mediated exceptions to the free ion model. AB - The accumulation of Cd in durum wheat (Triticum turgidum) roots from hydroponic solutions, with the proportion of total Cd (8.9-445 nM Cd) as Cd2+ varied by the addition of citrate, was determined to test the free-ion model (FIM) of metal bioavailability for higher plants. Calcium, Mg, and K were also varied. Citrate enhanced root-Cd accumulation at higher Cd2+ concentrations but not lower relative to the same Cd2+ concentrations in solutions containing 0 mM citrate. Elevating Ca2+ and Mg2+ concentrations in the citrate solution to the same as those in control solutions alleviated some of the citrate-mediated enhancement but not all. Solutions containing 66% less Ca or Mg than control but the same Cd2+ concentration and no citrate also resulted in increased root Cd. Elevated K+ did not influence Cd accumulation. Regression relationships between root-Cd accumulation and total Cd in solution were similar for the control and pooled amended solutions, whereas they were different for root-Cd accumulation and solution Cd2+. These results contribute to the growing body of evidence that the FIM alone is likely insufficient to predict plant accumulation of metals from soils, although it may be a useful probe for the mechanistic bases of metal bioavailability. PMID- 12729230 TI - Dietary zinc reduces uptake but not metallothionein binding and elimination of cadmium in the springtail, Orchesella cincta. AB - In metal-polluted environments, high concentrations of metals such as zinc, cadmium, copper, and lead are present. Many studies have been conducted to determine uptake and binding of the nonessential, toxic metal cadmium. Effects of essential metals on this uptake and binding remain unclear. The aim of the present study was to determine the effect of zinc on cadmium kinetics and metallothionein induction in the springtail Orchesella cincta. Exposure experiments were conducted with cadmium only, zinc only, and cadmium + zinc. Metallothionein was isolated to determine zinc binding to or induction of this cadmium-binding protein. In animals exposed to cadmium only and to cadmium + zinc, metallothionein was induced. In animals exposed to zinc only, no metallothionein was induced, and zinc did not bind to the metallothionein induced by cadmium. Cadmium uptake rate was significantly lower in animals exposed to cadmium + zinc than in animals exposed to cadmium only. No significant pattern of uptake and elimination of zinc was observed. Zinc concentrations in the animals were not significantly different for the different exposures, demonstrating that this element is regulated by O. cincta. These results suggest a protective role of zinc in the uptake of cadmium but no interference of zinc with the internal cadmium distribution and elimination. PMID- 12729229 TI - Influence of aging on copper bioavailability in soils. AB - Because of long-term chemical processes, metal bioavailability in field soils decreases with time. Metal toxicity may, therefore, be overestimated if toxicity data with freshly spiked soils are used to derive soil quality criteria, a current practice. In the present study, effects of the long-term processes, called aging, on copper partitioning and ecotoxicity are investigated. Twenty five field soils contaminated by copper runoff from bronze statues and 25 uncontaminated control soils sampled at 5-m distance from these statues were collected in Flanders (Belgium). The soils were selected so that parameters affecting copper bioavailability (pH, cation-exchange capacity, organic matter content, etc.) varied considerably. To assess the effect of aging on copper toxicity, control soils were spiked at total copper concentrations comparable to those of historically contaminated soils. Pore-water copper concentrations and 0.01 M CaCl2-extracted copper concentrations were significantly higher in freshly spiked soils compared to contaminated field soils. However, this could be a pH effect, because pH decreased after spiking. Acute toxicity to Enchytraeus albidus (14 d) as well as chronic toxicity to Folsomia candida (28-d reproduction) and Trifolium pratense (14-d growth) indicated a dose-response relationship between copper toxicity and pore-water copper concentration or the CaCl2-extracted copper fraction. PMID- 12729231 TI - Using a freshwater amphipod in situ bioassay as a sensitive tool to detect pesticide effects in the field. AB - In situ testing represents an alternative to conventional laboratory toxicity testing of field samples. Juvenile Paramelita nigroculus (Crustacea: Amphipoda) were exposed in situ in two rivers downstream of fruit orchard areas in the Western Cape, South Africa. Exposure took place during six time intervals (3-7 d), of which three represented the first rainfall-induced edge-of-field runoff events of the wet season, which was about two months after the last pesticide application. Survival rates were significantly reduced (35-70%) during runoff events 1 and 2 at both sites. No difference was observed from the respective no runoff survival rate (>90%) during event 3. Peak levels of total insecticides (azinphosmethyl, chlorpyrifos, endosulfan, and prothiofos) in samples taken with water level-triggered samplers during the runoff events were between 0.03 and 0.26 microg/L in filtered water and between 305 and 870 microg/kg in suspended particles during runoff events 1 and 2 and only up to 0.01 microg/L and 101 microg/kg during event 3. Total suspended solids (TSS) varied between 400 and 700 mg/L during all three runoff events but never exceeded 65 mg/L during no-runoff time intervals. A laboratory experiment revealed that uncontaminated TSS levels of 1,500 mg/L during a 7-d exposure caused insignificant mortality (<2.5%) in P. nigroculus. No acute toxicity was observed in standard 48-h toxicity tests with juvenile Daphnia pulex using the filtered water samples taken during runoff and no-runoff conditions. It is concluded that the observed mortalities were caused by particle-associated pesticides and that the present amphipod in situ bioassay represents a sound and sensitive tool to detect runoff-related insecticide effects under field conditions. PMID- 12729232 TI - Family physicians and osteoporosis. Meeting the challenge. PMID- 12729233 TI - Fewer medical students selecting family medicine. Can family practice survive? PMID- 12729234 TI - Understanding "beings" is the challenge. PMID- 12729235 TI - Who is that woman? PMID- 12729236 TI - Jumping off the hormone bandwagon. PMID- 12729238 TI - Practice tips. Preventing hip fractures in elderly patients. PMID- 12729237 TI - Anorexia nervosa during pregnancy. AB - QUESTION: A 22-year-old patient in my clinic was diagnosed with anorexia nervosa (AN) 7 years ago. She is now married and planning her first pregnancy. She is still underweight. What should she expect during pregnancy, and are there any implications for her unborn baby? ANSWER: Women with AN are at higher risk of complications during pregnancy, mainly because of low body weight. Apgar scores and birth weights of infants born to mothers with AN have been found to be significantly lower than those of infants born to healthy women. Rates of cesarean delivery, postnatal complications, and postpartum depression are higher among mothers with AN. Complications include hypothermia, hypoglycemia, infections, and increased rates of perinatal death. It is important to ensure appropriate intake of not only calories and proteins but also micronutrients, such as folic acid, to prevent neural tube defects. PMID- 12729239 TI - Intra-articular hyaluronic acid injections. Are they effective treatment for osteoarthritis of the knee? PMID- 12729240 TI - Advances in osteoporosis therapy. 2003 update of practical guidelines. AB - OBJECTIVE: To review evidence for current therapies for postmenopausal osteoporosis and to establish practical guidelines for management of osteoporosis by family physicians. QUALITY OF EVIDENCE: MEDLINE was searched from January 1990 to January 2003. Articles retrieved were graded by level of evidence (I to III). Recommendations for diagnosis and therapy were based on evidence from randomized controlled trials and meta-analyses. MAIN MESSAGE: Osteoporosis is treatable. Early diagnosis and intervention is recommended. After excluding secondary causes of osteoporosis, physicians should advise patients to take appropriate calcium and vitamin D supplementation. Those with osteopenia at risk of fractures and those with established osteoporosis need additional therapy. CONCLUSION: Approved pharmacologic therapies include alendronate, risedronate, raloxifene, calcitonin, cyclical etidronate, and hormone replacement therapy. Family physicians can help with early diagnosis and intervention and should discuss lifestyle modification with patients. PMID- 12729241 TI - What to do if an initial antidepressant fails? AB - OBJECTIVE: To provide family physicians with practical ways of managing depressed patients responding insufficiently to initial antidepressant treatment. QUALITY OF EVIDENCE: A search of MEDLINE and relevant bibliographies showed most studies could be categorized as level III evidence. Few well controlled studies (eg, level I evidence) specify treatment of next choice in rigorously defined treatment-refractory depression (TRD). MAIN MESSAGE: Failure to achieve and sustain full symptom remission affects relatively few treated depressed patients. Most chronically depressed people are not absolutely resistant but are relatively resistant to treatment; they fail to achieve the goals of treatment because of improper diagnosis or insufficient treatment application. The literature on TRD has largely focused on medication strategies; fewer studies investigated psychosocial approaches. The best established augmentation strategies are lithium salts and triidothyronine (T3). Combination antidepressants have become clinical psychiatrists' preferred treatment, despite limited evidence. Electroconvulsive therapy (ECT) remains a feasible option for TRD, but response rates are poor among people with TRD. High relapse rates after ECT remain a serious and common clinical dilemma. CONCLUSION: Family physicians should familiarize themselves with some new strategies to modify inadequate response to initial antidepressant treatment. PMID- 12729242 TI - How are family physicians managing osteoporosis? Qualitative study of their experiences and educational needs. AB - OBJECTIVE: To explore family physicians' experiences and perceptions of osteoporosis and to identify their educational needs in this area. DESIGN: Qualitative study using focus groups. SETTING: Four Ontario sites: one each in Thunder Bay and Timmins, and two in Toronto, chosen to represent a range of practice sizes, populations, locations, and use of bone densitometry. PARTICIPANTS: Thirty-two FPs participated in four focus groups. Physicians were identified by investigators or local contacts to provide maximum variation sampling. METHOD: Focus groups using a semistructured interview guide were audiotaped and transcribed. The constant comparative method of data analysis was used to identify key words and concepts until saturation of themes was reached. MAIN FINDINGS: Family physicians order bone densitometry and try to manage osteoporosis appropriately, but lack a rationale for testing and are confused about management. Participants' main concern was clinical management, followed by disease prevention and their educational needs. CONCLUSION: Family physicians are confused about how to manage osteoporosis. To reduce the burden of illness due to osteoporosis, educational interventions should be tailored to family physicians' needs. PMID- 12729243 TI - [Residency in family medicine: problems and solutions]. AB - OBJECTIVE: To identify the problems encountered by residents during training and the solutions they proposed. DESIGN: Qualitative study. SETTING: Family practice program at Sherbrooke University. PARTICIPANTS: Sixty-eight residents in the Family practice program between 1999 and 2000. METHOD: We used the nominal group technique with four groups of participants. Then we held five 3-hour discussion groups to explore difficulties and solutions in depth. Finally, data were validated with a written questionnaire. MAIN FINDINGS: At least 20% of respondents confirmed 65 difficulties and 61 solutions. Three major themes emerged: the many demands of the residence program, residents' personal experience of the program, and the lack of time for personal life. CONCLUSION: Publication of these results could help residents develop proactive adaptive strategies to deal with the stress of their training programs. Certain adjustments will be made by the program administration to make it easier for residents to manage their stress. PMID- 12729244 TI - Canadian consensus on osteoporosis. Preventing osteoporosis among postmenopausal women. PMID- 12729245 TI - Clinical guidelines for depressive disorders. Summary of recommendations relevant to family physicians. PMID- 12729247 TI - Metals in perspective. PMID- 12729246 TI - Can lack of communication kill? PMID- 12729248 TI - Nickel toxicology in retrospect. PMID- 12729249 TI - Workers' health in Russia: current problems and analysis. PMID- 12729250 TI - The role of chromatin damage in nickel-induced carcinogenesis. A review of recent developments. AB - Over the last years, we have been testing a hypothesis that molecular mechanisms of nickel-induced carcinogenesis include interactions of this metal with major chromatin components; DNA, histones, and protamines. Our investigations using synthetic peptide models have resulted in identification of nickel-binding sites in core histones H3 and H2A and in protamine P2. These are: the internal -Cys110 AIH- motif in histone H3: the C-terminal-E121-SHHKAKGK "tail" motif in histone H2A; and the N-terminal RTH- motif in protamine P2. Ni(II) bound to the H3 and P2 motifs enhances oxidative DNA base damage by H2O2. In contrast, Ni(II) complex with the H2A "tail" is not redox active. However, at pH 7.4, it undergoes hydrolysis yielding a new complex, Ni(II)-SHHKAKGK, reactive with H2O2 and capable of mediating DNA oxidation. The "tail" cutting of H2A has also been observed in cells cultured with Ni(II). In Ni(II) complex with the protamine P2 peptides, H2O2 causes degradation of the metal-binding His3 and the distant Tyr8 residues. This site-specificity results from a long-range structuring effect of Ni(II) on its protamine ligand. In conclusion, Ni(II) binding to some chromatin proteins in somatic and sperm cells may result in oxidative and structural damage to the proteins and DNA. These effects may alter the fidelity of DNA replication and gene expression and thus facilitate carcinogenesis, including paternally mediated cancer in the progeny. PMID- 12729251 TI - Epidemiology of nickel allergy. PMID- 12729252 TI - Lung cancer incidence among Norwegian nickel-refinery workers 1953-2000. AB - Among workers employed at a nickel refinery in Norway between 1910 and 1977 an elevated risk of lung cancer has been demonstrated. A dose-related effect from nickel exposure has been identified, with the strongest gradient for water soluble nickel. This pattern was recently confirmed in a nested case-control study with adjustment for smoking and potential occupational confounders. In the present study, updated cancer data were used to explore the risk by duration of work at the refinery and by exposure to different forms of nickel. Comparisons were made with the national male population (standardised incidence ratios) as well as internal reference groups (Poisson regression) under adjustment for age and smoking. The results confirmed earlier findings of a strong dose-related risk dependent on duration of work in production departments and cumulative exposure to nickel, most clearly seen for water-soluble nickel. Only slightly elevated risks were found among the unexposed and in the group with no experience from production or maintenance work. The risk associated with exposure to nickel chloride was similar to that for nickel sulfate. Analyses restricted to men exposed after 1967, with estimates based on personal monitoring of nickel in the breathing zone, showed the same risk pattern as for earlier years. Elevated lung cancer incidence was even suggested for workers with their first employment after 1978 when a lot of high exposure jobs were abandoned. The combined effect of exposure to nickel and smoking seemed to be in agreement with a multiplicative risk pattern. PMID- 12729253 TI - Cigarette smoking and nickel exposure. AB - The tobacco plant contains nickel and several other toxic metals, most probably absorbed from the soil, fertilizing products or from pesticides. It has been stated that nickel in a burning cigarette might form the volatile, gaseous compound, nickel tetracarbonyl, and thereby be introduced into the respiratory tract. Accordingly, the main objective of the present study was to find out if nickel content in inhaled smoke from ordinary cigarettes and nickel-contaminated cigarettes handmade by nickel process workers might be a supplementary source of nickel exposure to cigarette smoking process workers leading to additional risk of occupational respiratory cancer in these workers. Blood and urine samples from 318 randomly selected employees from Falconbridge Nickel Refinery in Kristiansand, Norway, allocated to 197 smokers and 121 non-smokers, were analysed for nickel content. Nickel quantities in tobacco from various cigarette brands, from nickel-contaminated cigarettes made by process workers or from cigarettes added known amounts of various nickel salts were analysed before being smoked. The cigarettes were smoked in a smoking machine device applying an electrostatic filter. Blood and urine, tobacco, ash and precipitates in the filter from the main stream smoke of the cigarettes were analysed for nickel quantities by atomic absorption spectrometry methods as previously described by the authors. The nickel concentrations in blood plasma and urine were quite similar among smokers and non-smokers, 6.2 and 48.1 microg L(-1) in smokers, and 6.4 and 50.5 microg L( 1) in non-smokers respectively. We recovered 1.1% or even less of nickel in the mainstream smoke after smoking the entire cigarettes without leaving any butt. Most of the tobacco nickel was recovered in the ash. We conclude that the inhaled nickel in the working atmosphere is probably the main source of the nickel exposure to the respiratory tract in these workers. It remains to be determined why cigarette smoking still seems to be a decisive cofactor in the development of respiratory tract cancer in nickel workers. PMID- 12729254 TI - X-ray diffraction spectrometric analysis of nickel refinery aerosols, process materials and particulates isolated from worker lung tissues. AB - Results are reported of X-ray diffraction analysis of extracts derived from lungs of two nickel refinery workers and of three stationary air samples collected inside a nickel refinery. Since environmental samples from the 1950s and 1960s do not exist, two archived production control samples from that period were also analyzed. Because nickel has been found in respiratory tissue of workers retired for more than twenty years, it was likely that the residual nickel compounds must be rather insoluble. Preliminary surveys showed that sulfur was not present in the lung tissue deposits and thus water-soluble and sulfidic nickel were therefore extracted from the ten process samples before the X-ray analysis. A common compound that was found in all 10 samples was trevorite. This is a spinel type mineral, much like magnetite where the divalent iron is replaced by nickel. It may be formed when trivalent iron reacts with nickel at 1100 degrees C. It has magnetic properties and is very insoluble. Samples from the lungs were obtained by burning off the organic tissue at 630 degrees C. Due to a relatively high detection limit for the X-ray diffraction technique, we were initially not able to detect any mineral nickel compound. But when particles extracted with a magnet were analyzed, a very clear diffraction pattern of trevorite was identified. The main residue after the magnetic separation had a low concentration of nickel (4 microg g(-1)), which suggests that trevorite was the dominating, if not the only, nickel compound present. In addition, chemical analyses were performed on 13 tissue samples from one single lung; one from each main bronchus, two from each lobe, and an additional one from the lower right lobe. Statistical testing showed a highly significant correlation between the five elements determined: Ni, Co, Cu, Fe and Cr. This suggests that these metals are isomorphous and substitute for each other in the mineral structure. These results may indicate that the nickel left in the lungs some years after exposure is trevorite, and may be biologically inert. However, further speciation of nickel compounds in the lungs seems warranted, and trevorite should be tested for its potential toxic effects. PMID- 12729255 TI - GeneChip analysis of signaling pathways effected by nickel. AB - The carcinogenicity of nickel compounds has been shown in numerous epidemiological and animal studies. Carcinogenesis is generally considered as a multistep accumulation of genetic alterations. Nickel, however, being highly carcinogenic is only a weak mutagen. We hypothesize that nickel may act by modulating signaling pathways, and subsequently by reprogramming transcription factors. Insoluble nickel is considered to be more carcinogenic than soluble. In this study using GeneChip technology we compared changes in gene expression caused by soluble and insoluble nickel compounds. We found that both soluble and insoluble nickel compounds induce similar signaling pathways following 20 h of in vitro exposure. For example, both nickel compounds activated a number of transcription factors including hypoxia-inducible factor I (HIF-1) and p53. The induction of these important transcription factors exerts potent selective pressure leading to cell transformation. The obtained data are in agreement with our previous observations that acute nickel exposure activates HIF-1 and p53 transcription factors and in nickel-transformed cells, the ratio of HIF-I activity to p53 activity was shifted towards high HIF-I activity. The activation of the same signaling pathways by soluble and insoluble nickel compounds suggested that both nickel compounds have similar carcinogenic potential in vitro. PMID- 12729256 TI - Impact of mining and refining on the distribution and accumulation of nickel and other heavy metals in sediments of subarctic Lake Kuetsjarvi, Murmansk Region, Russia. AB - Research on the influence of the activities of Pechenganickel Mining and Metallurgical Company on sediment heavy-metal geochemistry of the subarctic Lake Kuetsjarvi (north-western Russia) are described. It is estimated that during 60 years of mining/refining activity, 310 t of Ni, 120 t of Cu, 14 t of Co, 19 t of Zn, 0.087 t of Cd, 0.78 t of Pb and 0.053 t of Hg have accumulated in the lake sediments. The latter can be a source of secondary pollution and represent a danger for the lake ecosystem. The sedimentation rate in the lake is estimated to be within the range of 1.5-3 mm year(-1). The average concentrations of Ni, Cu, Hg and Co in superficial sediments have increased 25, 14, 11 and 5 times, respectively in the last century. PMID- 12729257 TI - Morphometric and metabolic indicators of metal stress in wild yellow perch (Perca flavescens) from Sudbury, Ontario: a review. AB - Eighteen lakes studied near Sudbury and across Northeastern Ontario (Canada) over a five-year period provided a wide contamination gradient of cadmium (Cd), copper (Cu) and other metals such as nickel (Ni) and zinc (Zn). All were inhabited by yellow perch (Perca flavescens), which was sometimes the only species present. Liver Cd and Cu concentrations were monitored in these lakes, in some cases for several consecutive years and for multiple seasons. This data suggests that yellow perch from clean to mildly-contaminated environments loosely control their hepatic Cu concentrations between 7 and 50 microg g dry weight(-1), and a threshold of 50 microg g dry weight(-1) is suggested as the normal range of homeostatic control. Similar data collected by others support this value. Liver Cd concentrations appeared more variable among lake samples, but consistently remained below 10 microg g dry weight(-1) in clean to mildly-contaminated lakes, also supported by data collected elsewhere. Condition factors allowed the discrimination between clean and polluted yellow perch, a conclusion consistent with data for the same species collected in the Rouyn-Noranda area (Quebec, Canada). Values of weight-to-length scaling coefficient lower than 3.0 also discriminated between clean and metal-polluted yellow perch. Finally, three studies indicated that chronic metal exposure can lead to an impairment of aerobic capacities in wild yellow perch, as indicated by lower muscle activity of citrate synthase (CS), aerobic swim performance and respiration rate. We propose that the combination of liver metal concentrations, scaling coefficient, condition factor and an indicator of physiological impairment such as muscle CS activity can provide a suitable range of parameters to adequately assess the effects of metal contamination on the health of yellow perch. Although yellow perch are ubiquitous in North America, this approach can potentially be applied to other small fish species more suitable to other study areas. PMID- 12729259 TI - Human dietary intake and excretion of dioxin-like compounds. AB - Human dietary intake and excretion of polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), and biphenyls (PCBs)--collectively referred to as dioxin like compounds (DLCs)--were investigated. Two groups of seven subjects were studied during 1999 and 2000: one aged 23.9 +/- 4.5 years, the other aged 49.4 +/ 5.2 years. For each subject, two week-long experiments were held. In one, omnivorous diets were administered, whilst in the other, a vegan diet was studied. While exposures via the omnivorous diets exceeded those via the vegan diet on a sigmaWHO-TEQ basis; for some subjects sigmaPCB exposures were comparable in both diets, implying that plant-based foods can make an appreciable contribution to exposure to sigmaPCB. For all subjects, the average dietary exposure during the omnivorous trial to PCDD/Fs and PCBs combined (expressed as WHO-TEQ) was--at 1.09 pg kg(-1) bw d(-1)--lower than recent UK "food-basket" estimates. For the same diet, the average sigmaPCB exposure for all subjects was- at 5.01 microg person(-1) d(-1)--higher than recent UK "food-basket" estimates, but consistent with recent estimates for other industrialised countries. Net absorption/excretion but not faecal excretion rates of DLCs were related to dietary intakes. Furthermore, excretion rates of the most persistent DLCs--i.e. PCB #s 138, 153, and 1,2,3,6,7,8-HxCDD--were statistically significantly greater for the older subjects. Combined, these data imply that excretion rate is dependent on body burden, and that the majority of DLCs in human faeces arise from endogenous excretion. PMID- 12729260 TI - Dietary exposure to xenoestrogens in New Zealand. AB - Continuing evidence of the feminising effects of xenoestrogens on a range of wildlife species increases the need to assess the human health risk of these estrogen mimics. We have estimated the exposure of New Zealand males, females and young men to a range of naturally occurring and synthetic xenoestrogens found in food. Only estrogenic compounds that act by interaction with the estrogen receptor have been included. Theoretical plasma estrogen activity levels were derived from estrogen exposure estimates and estrogenic potency data. Theoretical plasma levels were compared with published data for specific xenoestrogens. There was surprisingly close agreement. Xenoestrogenicity from dietary intake was almost equally attributed to naturally occurring and synthetic xenoestrogens. Relative contributions for a male, for example were isoflavones (genistein and daidzein) (36%) and bisphenol A (34%) with smaller contributions from alkyl phenols (18%) and the flavonoids (phloretin and kaempferol) (12%). It is suggested that dietary xenoestrogens might have a pharmacological effect on New Zealand males and postmenopausal women, but are unlikely to be significant for pre-menopausal women. PMID- 12729258 TI - Molecular mechanisms of nickel carcinogenesis: gene silencing by nickel delivery to the nucleus and gene activation/inactivation by nickel-induced cell signaling. AB - We have summarized the molecular and cellular events involved in nickel (Ni) compound induced carcinogenesis. The major hypothesis for nickel carcinogenic action has involved the ability of the Ni compound to deliver high concentrations of Ni intracellularly, enter the nucleus and interact with chromatin. Ni has been found to selectively damage heterochromatin, and a major action of Ni is its ability to silence the expression of genes located near heterochromatin by inducing a loss of histone H4 and H3 acetylation and DNA hypermethylation. When Ni silences critical genes, such as tumor suppressor genes, the cell is altered to a greater state of neoplastic transformation. The carcinogenic hazard of Ni compounds has been directly related to the ability of that Ni compound to raise the intracellular Ni ions. The mechanisms of Ni-induced gene silencing will be discussed. However, recently it has been found that soluble Ni ions can interact with the cell surface receptors and activate cell signaling resulting in the induction of a variety of cellular genes. In particular, the Ca and hypoxia inducible factor pathway is activated in all cells exposed to soluble Ni ions. In the case of HIF-1 induction, a cell is now equipped with the expression of a variety of genes that will allow the cell to survive the lack of oxygen and thus should enable a previously initiated cancer cell to progress into a full malignant state and metastasize. These new findings support the view that soluble Ni ions exhibit carcinogenic potential by activating cell promotion and lend strength to the epidemiological data showing soluble Ni to be associated with cancer risk in Ni refinery workers. PMID- 12729261 TI - Occupational exposure to fluorinated hydrocarbons during refrigeration repair work. AB - This study describes refrigeration repair workers' occupational exposures to halogenated refrigerants, focusing on difluorochloromethane (HCFC 22), tetrafluoroethane (HFC 134a) and a mixture of tri-, tetra- and pentafluoroethane (R404A) in 30 work operations. Unlike earlier reported studies, the present study includes working procedures involving welding in order to measure possible occupational exposure to decomposition products. The measurements included hydrogen fluoride (HF), hydrogen chloride (HCl), phosgene (COCl2) and volatile organic compounds (VOC). The exposures were assessed during work operations on small-scale cooling installations like refrigerators and freezers. The repair workers' occupational exposures to refrigerants were moderate, and the major part of the exposures were associated with specific working procedures lasting for relatively short periods of time (<20 min). During these exposure events the concentrations were occasionally high (up to 42434 mg m(-3)). Although welding operations lasted only for short periods of time, HF was detected in 9 out of 15 samples when HCFC 22, HFC 134a or R404A had been used. Hydrogen chloride was detected in 3 out of 5 samples in air polluted with HCFC 22. Phosgene was not detected. A large number of VOCs in various concentrations were found during welding. Except for the applied refrigerants, halogenated compounds were only found in one sample. PMID- 12729262 TI - Impact on blood Pb levels of maternal and early infant feeding practices of First Nation Cree in the Mushkegowuk Territory of northern Ontario, Canada. AB - Exposure to Pb in-utero and in infancy has been associated with cognitive risk, even at low blood Pb levels of 0.48-0.96 micromol L(-1). Based on the hypothesis that wild game consumed by First Nation women of Mushkegowuk Territory contains Pb that might be transferred through blood or breast-milk to the fetus or infant, the study's objectives were to describe: (1) Pb in maternal and cord blood at birth and infant blood at 4 months, and (2) dietary influence on Pb status. Cord and maternal Pb were 0.10 +/- 0.08 and 0.11 +/- 0.06 micromol L(-1) (x +/- SD), respectively, and were significantly correlated (r = 0.77, p < 0.0001, n = 70), as was infant blood Pb (0.08 +/- 0.05 micromol L(-1)) with matched cord blood (r = 0.65, p < 0.0001, n = 30). Two cord blood samples (3%) were above 0.48 micromol L(-1). Pb in breast-milk at 0.010 +/- 0.008 micromol L(-1) (n = 25), was significantly lower than Pb in commercial formula or evaporated milk-based feedings (range 0.02-0.05 micromol L(-1), p < 0.05), and correlated with matched maternal blood Pb (r = 0.55, p < 0.005). However, in a sub-sample of infants (n = 31), blood Pb was similar for breastfed and formula-fed groups, though above the evaporated milk-fed group (p < 0.05). Maternal consumption of wild fowl, mammals and fish, estimated from the previous year, provided, respectively, 128 +/- 124, 46 +/- 68 and 8 +/- 13 MJ annum(-1). Traditional animal food intake, especially wild fowl, correlated significantly with cord blood Pb (Spearman rank correlation, p = 0.017). Although blood and milk Pb levels were largely within acceptable ranges, the presence of some elevated levels and association between blood Pb and traditional game consumption may reflect the legacy of using lead containing ammunition. PMID- 12729263 TI - Increased levels of bacterial markers and CO2 in occupied school rooms. AB - Our group previously demonstrated that carbon dioxide (CO2) levels in heavily occupied schools correlate with the levels of airborne bacterial markers. Since CO2 is derived from the room occupants, it was hypothesized that in schools, bacterial markers may be primarily increased in indoor air because of the presence of children; directly from skin microflora or indirectly, by stirring up dust from carpets and other sources. The purpose of this project was to test the hypothesis. Muramic acid (Mur) is found in almost all bacteria whereas 3-hydroxy fatty acids (3-OH FAs) are found only in Gram-negative bacteria. Thus Mur and 3 OH FA serve as markers to assess bacterial levels in indoor air (pmol m(-3)). In our previous school studies, airborne dust was collected only from occupied rooms. However, in the present study, additional dust samples were collected from the same rooms each weekend when unoccupied. Samples were also collected from outside air. The levels of dust, Mur and C10:0, C12:0, C14:0, and C16:0 3-OH FAs were each much higher (range 5-50 fold) in occupied rooms than in unoccupied school rooms. Levels in outdoor air were much lower than that of indoor air from occupied classrooms and higher than the levels in the same rooms when unoccupied. The mean CO2 concentrations were around 420 parts per million (ppm) in unoccupied rooms and outside air; and they ranged from 1017 to 1736 ppm in occupied rooms, regularly exceeding 800-1000 ppm, which are the maximum levels indicative of adequate indoor ventilation. This indicates that the children were responsible for the increased levels of bacterial markers. However, the concentration of Mur in dust was also 6 fold higher in occupied rooms (115.5 versus 18.2 pmole mg( 1)). This further suggests that airborne dust present in occupied and unoccupied rooms is quite distinct. In conclusion in unoccupied rooms, the dust was of environmental origin but the children were the primary source in occupied rooms. PMID- 12729264 TI - Public perceptions of air quality and quality of life in urban and suburban areas of London. AB - A comparative study was undertaken at two different sites (one urban, one suburban) in Greater London in order to examine whether there was a relationship between publicly available air quality data and the public's perception of air quality. Perceptions of air quality and its role as a potential quality of life indicator was also investigated. A total of 200 people were interviewed in Wood Green and Wimbledon in 1999 and air quality data were obtained simultaneously from the nearest appropriate monitoring station. The study has enabled a comparison between the public's perception of air quality and the actual monitored concentration values. The survey results reveal how seriously the public regards air pollution from road traffic in terms of their quality of life and identifies the attitudes of the public to nuisance from road traffic. The results reveal that the public's perception of air quality is not a reliable indicator of the actual levels of air pollution in their area. The results also revealed that air pollution issues generated as a result of road traffic are of high importance in terms of people's quality of life when compared to other aspects of their quality of life. The study revealed that residents in the urban area (Wood Green) were more disturbed by road traffic than residents in the suburban area (Wimbledon). Out of all the disturbances listed, it was found that residents were more disturbed by vehicle-derived fumes, dust and dirt than other aspects of road traffic related nuisance. The TELETEXT/CEEFAX service on air quality was little used by the public as a means of obtaining information on air pollution. PMID- 12729265 TI - Short-term variation in air quality associated with firework events: a case study. AB - The effect of fireworks on air quality was assessed from the ambient concentrations of various air pollutants (SO2, NO2, PM10 and TSP) during Diwali festival in Hisar city (India), in November 1999. The extensive use of fireworks was found to be related to short-term variation in air quality. During the festival the concentration of SO2 was observed to be increased approximately 10 fold at few sites, whereas the concentrations of NO2, PM10 and TSP increased 2-3 times, compared to the data collected on a typical winter day in December 1999. The maximum NO2 concentration was observed a day after the festival. The diurnal pattern of the above pollutants showed a slight increase in the night. The levels of these pollutants observed during Diwali were found to be moderately high, which can be associated with serious health impacts. PMID- 12729266 TI - Ion-soot interaction: a possible mechanism of ion removal in aircraft plume. AB - The phenomenon of the ion-soot interaction in the aircraft plume at the ground conditions is investigated. The ion-soot attachment coefficients, taking into account the polarization of the soot particles in the ion electric field, are calculated. It is shown that the ion-soot attachment may play the important role in the evolution of the ion concentrations in the plume. Comparison of the model results with the ground-based measurements for the ion depletion along the plume demonstrates that the concentration of the positive and negative ions at the nozzle exit for these observations is close to 1.2 x 10(8) cm(-3). PMID- 12729267 TI - Estrogenic chemicals and estrogenic activity in leachate from municipal waste landfill determined by yeast two-hybrid assay. AB - Estrogenic activity and estrogenic chemicals in landfill leachate were investigated by yeast two-hybrid assay and chemical analysis. Leachate sample extracted by liquid-liquid extraction with dichloromethane at pH 7.0 showed a higher dose-response curve than sample extracted at pH 3.0. or than sample extracted by solid phase extraction at either pH 7.0 or 3.0. The fraction extracted at pH 3.0 specifically inhibited not only growth of yeast but also estrogenic activity in this assay, suggesting that it contained anti-estrogenic chemicals. The greatest contributor to estrogenic activity among the chemicals identified in leachate extract was bisphenol A, with an estimated contribution ratio of 84%. The contribution ratios of 4-nonyl phenol (4-np) and 4-tert-octyl phenol (4-t-op) were estimated at 1.0%, and 0.1%, respectively, while natural estrogens such as 17beta-estradiol or estrone were below detection limit, so that their contribution ratio was estimated at no more than 10%. The estrogenic activity of leachate was decreased by aeration treatment alone after 7 days, and was no longer detected after 22 days. Concentrations of bisphenol A, 4-np and 4-t op likewise decreased with aeration. PMID- 12729268 TI - Chloroacetic acids in European soils and vegetation. AB - Trichloroacetic acid (TCA) and dichloroacetic acid (DCA) are possible minor atmospheric degradation products of perchloroethylene and trichloroethylene, respectively. These acids may be wet- or dry-deposited from the atmosphere to land surfaces and hence possibly affect plant growth. However, the existing database on TCA levels in soil is limited to a few studies carried out in the late 1980's and the early to mid-1990's and it was concluded that there is a need for further measurements of concentrations of TCA and DCA in soils. In this study soil samples from 10 locations in 5 European countries, as well as vegetation samples, and a limited number of rainwater and air samples were collected and analysed for DCA and TCA to determine the concentrations of these compounds. An isotope dilution method using GC-MS was used for the determination of these acids in the samples. The method was briefly validated and the performance characteristics are presented. The results of the analysis of the soil samples show that the DCA and TCA concentrations in soil from different sites in Europe are more or less comparable, with the exception of Germany, especially Freudenstadt, where significantly higher TCA concentrations (up to 12 microg kg( 1) dw) were found. The average DCA and TCA concentrations in soil in this study were 0.25 +/- 0.12 and 0.64 +/- 1.40 microg kg(-1) dw, respectively. Generally, the concentration in soils from forest areas are about twice those from open-land areas. The DCA and TCA concentrations in vegetation samples ranged from 2.1 to 73 microg kg(-1) dw for DCA and from 4.7 to 17 microg kg(-1) dw for TCA. Thus, the concentrations in vegetation samples are 10-20 times higher than the soil concentrations. DCA and TCA concentrations in wet deposition samples and air samples collected in The Netherlands were 0.14 and 0.15 microg l(-1) for wet deposition samples and <0.5 and 0.7 ng m(-3) for air samples respectively. For these samples taken in The Netherlands, the estimated values for soil and air concentrations calculated from the wet deposition concentrations are in agreement with the concentrations measured in this study. PMID- 12729269 TI - Distribution of polychlorinated biphenyls (PCBs) and organochlorine pesticides in soils from the East Antarctic coast. AB - Concentrations of hexachlorobenzene (HCB), alpha-, beta- and gamma hexachlorocyclohexane (HCH) isomers, 6 o,p'-and p,p'-isomers of DDT and 28 PCB congeners have been measured in eleven soil samples and one lichen collected on the Eastern coast of Antarctica from 5 Russian stations. For samples with low concentrations of PCBs (range 0.20-0.41 ng g(-1) dry weight) and pesticides (0.86 4.69 ng g(-1) and 0.11-1.22 ng g(-1) dry weight for HCHs and DDTs, respectively), atmospheric long-range transport from Africa, South America or Australia was suggested as the sole source of contamination. The profile of PCB congeners was dominated by the more volatile tri-, tetra- and penta-PCBs congeners, thus supporting long-range transport hypothesis. Four samples contained moderate levels of PCBs (range 1.98-6.94 ng g(-1) dry weight) and variable concentrations of pesticides (gamma-HCH, p,p'-DDT and o,p'-DDT being the main contaminants). For samples with high concentrations of PCBs (range 90.26-157.45 ng g(-1)) and high concentrations of pesticides, the presence of high molecular weight PCB congeners such as: 153, 180, 187, 170 etc, strongly suggest a local source (biotic) of PCBs rather than atmospheric transport. It is likely that on a local scale, biotic focussing of pollutants, due to bird activities (nesting and excrement) can cause high contamination levels and become more significant than contaminant input via abiotic pathways. PMID- 12729271 TI - Geotrupine beetles (Coleoptera: Scarabaeoidea) as bio-monitors of man-made radioactivity. AB - Adults of the geotrupine beetle Anoplotrupes stercorosus (Coleoptera, Geotrupidae), a common European forest insect species, were used in the role of bio-monitors for mainly man-made radionuclides in a forest environment. Activities of 137Cs, 40K, 238Pu, (239+240)Pu, 90Sr and 241Am were studied. Samples originated from four areas in Poland, two from the north-east and two from the south of the country. The north-eastern areas were previously recognized as the places where hot particle fallout from Chernobyl took place. Results confirmed the differences in the activities between north-eastern and southern locations. Significant correlations were found between activities of 40K and 137Cs, and between activities of plutonium and americium isotopes. An additional study of the concentration of radionuclides within the bodies of beetles showed a general pattern of distribution of radioisotopes in the insect body. PMID- 12729270 TI - Monitoring of copper, arsenic and antimony levels in agricultural soils impacted and non-impacted by mining activities, from three regions in Chile. AB - This paper reports a comparative study of the concentration of three important environmental elements that are often found together in mineral deposits and then associated with mining activities; copper, arsenic and antimony. These elements were determined in 26 different agricultural soils from regions I, II and V in Chile, zones where the most important and biggest copper industries of this country are located. As background levels of these elements in soils have not been well established, in this study, both, impacted and non-impacted agricultural soils from different regions were considered. The relationships between the concentrations of these elements in soils were also examined. The concentration ranges for copper, arsenic and antimony were 11-530; 2.7-202 and 0.42-11 mg kg(-1) respectively. The copper concentrations in non-polluted soils from the north and central zone of Chile were similar. However, three sites from the north region have copper concentration as higher as 100 mg kg(-1), values that exceed the critical concentration for copper in soils. The concentration of arsenic and antimony in the north soils were higher than in non-impacted ones and, in the case of arsenic, greatly exceeded the world average concentration reported for this element in soils. The highest arsenic and antimony concentrations were found in Calama and Quillagua soils, two different sites in the Loa valley. The arsenic/antimony concentration ratio was higher in Quillagua soil. The high concentrations of three elements determined in impacted soils from region V (Puchuncavi and Catemu valleys) clearly shows the impact produced in this zone by the industrial and mining activities developed in their proximities. At Puchuncavi valley a clear decrease was observed in copper, arsenic and antimony concentrations in soils on the function of the distance from the industrial complex "Las Ventanas", and all concentrations exceeded the reported critical values for this matrix. Instead at Catemu valley, only the copper concentration was higher than this value. Statistically significant correlation was found for Cu-Sb in all soils; more significant Cu-As, Cu-Sb and Sb-As correlations were evaluated for soils from Puchuncavi and Catemu valleys, corroborating that high concentrations of copper, arsenic and antimony in these soils coming from the same pollution sources, the copper industry and the thermoelectric power plant. PMID- 12729272 TI - Recent trends of plutonium fallout observed in Japan: plutonium as a proxy for desertification. AB - Plutonium in monthly deposition samples collected in Tsukuba (the Meteorological Research Institute), Japan from 1990 to end of 2001 is reported, together with monthly plutonium deposition in Nagasaki and Yonaguni in 2000. The annual deposition of (239,240)Pu during the period from 1990 to 2001 shows no systematic interannual variation. However, monthly (239,240)Pu depositions show a typical seasonal variation with a maximum in spring season (March to April), which corresponds to seasonal cycle of soil dusts originating from the East Asian arid area. Plutonium isotopic ratios in the deposition samples suggest that significant amounts of the recent (239,240)Pu deposition observed in Japan are attributed to the resuspension of plutonium-bearing surface soil particles; resuspended plutonium originates from the East Asian arid areas. The recent increased tendency of (239,240)Pu content in residues in deposition samples may reflect desertification in the East Asian continent. PMID- 12729273 TI - Polonium 210Po in the phytobenthos from Puck Bay. AB - The aim of the work was to determine the 210Po content in phytobenthos species (seaweeds and angiosperms) from Puck Bay (southern Baltic). Alpha spectrometry was used to measure and calculate the activities and concentrations of polonium 210Po in the phytobenthos. The activity of 210Po in Puck Bay waters was determined to estimate the bioconcentration factors (BCF) of these plants. The 210Po concentration in water was estimated at 0.25 mBq dm(-3). The lowest polonium concentration in the phytobenthos was found in Cladophora rupestris (0.12 Bq kg(-1) wet wt.), the highest in Chara crinita (1.12 Bq kg(-1) wet wt.). Polonium is accumulated in these phytobenthos species; the bioconcentration factors (BCF) ranged from 450 to 4400. PMID- 12729274 TI - Analytical approaches to the determination of phosphorus partitioning patterns in sediments. AB - Three methods for phosphorus fractionation in sediments based on chemical extractions have been applied to fourteen aquatic sediment samples of different origin and characteristics. Two of the methods used different approaches to obtain the inorganic fractions. The Hieltjes and Lijklema procedure (HL) uses strong acids or bases, whereas the Golterman procedure (G) uses chelating reagents. The third one, the Standards, Measurements and Testing (SMT) protocol, was proposed in the frame of the SMT Programme (European Commission) which aimed to provide harmonisation and the validation of such methodologies. This harmonised procedure was also used for the certification of the extractable phosphorus contents in a sediment certified reference material (CRM BCR 684). Principal component analysis (PCA) was used to group sediments according to their composition and the three extraction methods were applied to the samples including CRM BCR 684. The data obtained show that there is some correlation between the results from the three methods when considering the organic and the residual fractions together. The SMT and the HL methods are the most comparable, whereas the G method, using a different type of reagent, yields different distribution patterns depending on sample composition. In relation to the inorganic phosphorus, the three methods give similar information, although the distribution between non-apatite and apatite fractions can be different. PMID- 12729275 TI - Determination of petroleum hydrocarbons and polycyclic aromatic hydrocarbons in sludge from wastewater treatment basins. AB - Screening by gas chromatography with flame ionization detection and gas chromatography-mass spectrometry has been carried out on sludge extracts of wastewater treatment basins. Soxhlet extraction with trichlorotrifluoroethane was applied. The yields for petroleum hydrocarbons and PAH recovery were high, usually in excess of 90%. The proposed investigations permit a quick assessment of petroleum pollutants in the environment. PMID- 12729276 TI - Statistical analysis of the physico-chemical data on the coastal waters of Cochin. AB - Measurements of temperature, salinity, dissolved oxygen, nitrogen as ammonia, nitrate and nitrite, and phosphate along with chlorophyll were carried out at three stations on the coastal waters of Cochin, south west India, at two-levels of the water column over a period of five years. The data set has been factorised using principal component analysis (PCA) for extracting linear relationships existing among a set of variables. A graphical display of the scores generated from the PCA was done by means of boxplots and biplots, which helped in the interpretation of the data. The major factors conditioning the system are related to the input of fresh water from the estuary of the Periyar river and the high organic load of the bottom sediment in the coastal area which results in a reducing environment, as reflected in the parameters of dissolved oxygen, ammoniacal-nitrogen and nitrite-nitrogen. Another factor which contributes to the variation in the system is related to the unloading activity in the port area. The present approach presents a logical way to interpret the complex data of the physico-chemical measurements. PMID- 12729277 TI - Seasonal variations of heavy metals in central Greenland snow deposited from 1991 to 1995. AB - To better assess the seasonality in the fallout of heavy metals to central Greenland, a continuous series of 68 snow samples has been collected at a remote site in the Summit area from a 2.7 m pit using ultraclean sampling procedures. This covers a continuous four year time period from spring 1991 to spring 1995. Co, Cu, Zn, Mo, Rh, Pd, Ag, Cd, Sb, Pt, Pb, Bi and U were determined using ultrasensitive inductively coupled plasma sector field mass spectrometry under clean room conditions. In addition we also determined Al by graphite furnace atomic absorption spectrometry and Na+, Ca2+, SO4(2-), MSA and oxalate by ion chromatography, species that will assist in the interpretation of the trace metal data. The data show pronounced inter- and intra-annual variations, with large differences in the amplitude of these variations for the element studied, with few clear seasonality patterns. Generally, high concentrations are observed in the spring snow layers, while much lower concentrations are typical of summer snow layers. Significant correlations are observed between Co, Cu, Zn, Ag and Sb, while Pt, Pd and Rh show no correlation with the other metals. Crustal enrichment factors show that while the crustal dust contribution is probably important for some metals for part of the year (spring), anthropogenic inputs are as important in many instances. Pronounced intra-annual variations are observed for some metals, in particular Pt. The variations observed for this metal parallel fairly closely changes in Russian Pt production, which points to emissions from smelters in the Russian Arctic as likely sources for Pt. PMID- 12729278 TI - Determination of ethanolamine, ethylene glycol and triethylene glycol by ion chromatography for laboratory and field biodegradation studies. AB - The determination of alkanolamines and glycols in groundwater and subsurface environments is essential for environmental assessment, remediation and monitoring for selected industrial sites. Monoethanolamine (MEA), ammonium, sodium, magnesium and calcium detection was performed using cation exchange chromatography (IC) with suppressed conductivity detection. Acetate, chloride, nitrite, nitrate, phosphate, sulfate and oxalate were monitored employing anion exchange chromatography with suppressed conductivity. Detection of ethylene glycol (MEG) and triethylene glycol (TEG) and ethanol was carried out using ion exclusion chromatography with pulsed amperometric detection. Effective determination of MEA, MEG and TEG in complex groundwater matrices without compound transformation offered improved monitoring capabilities. This study presents robust analytical tools for MEA, MEG and TEG determination in biodegradation studies. Using ion chromatography offered significant advantages for the analyses of groundwater samples and laboratory bioreactor monitoring. PMID- 12729279 TI - Occurrence and persistence of perfluorooctanesulfonate and other perfluorinated surfactants in groundwater at a fire-training area at Wurtsmith Air Force Base, Michigan, USA. AB - Various formulations of fire-extinguishing materials, including aqueous film forming foams (AFFFs), were used as part of fire-training exercises conducted at Wurtsmith Air Force Base (WAFB) in northeastern Michigan from the 1950s until the base was decommissioned in 1993. As a result of past fire-training exercises, AFFF-laden wastewater containing fuels, solvents, and other materials directly entered groundwater without prior treatment. Perfluorinated surfactants are key components in some AFFF formulations. In this study, groundwater was analyzed for perfluoroalkanesulfonates and perfluorocarboxylates. Perfluoroalkanesulfonates were directly detected using negative-ion electrospray ionization mass spectrometry. Derivatized perfluorocarboxylates were detected using electron impact gas chromatography-mass spectrometry. Groundwater from wells around fire training area FTA-02 at WAFB contained four perfluorinated surfactants ranging in concentration from 3 to 120 microg L(-1): perfluorooctanesulfonate (PFOS); perfluorohexanesulfonate; perfluorooctanoate; and perfluorohexanoate. This is the first report demonstrating that PFOS, recently shown to be toxic to organisms ranging from zooplankton to primates, is still present in groundwater in measurable quantities five or more years after its last known use. PMID- 12729280 TI - Monitoring of the three organophosphate esters TBP, TCEP and TBEP in river water and ground water (Oder, Germany). AB - The behaviour of the three organophosphate esters tributyl phosphate (TBP), tris(2-chloroethyl)phosphate (TCEP) and tris(2-butoxyethyl)phosphate (TBEP) during infiltration of river water to ground water has been investigated. The monitoring site is the Oder River and the adjacent Oderbruch aquifer. From March 2000 to July 2001, 76 ground water samples from monitoring wells located close to the Oder River and nine river water samples were collected. Additionally, influent and effluent samples from local waste water treatment plants, one sample of rain water and samples of roof runoff were collected. All samples were analysed by solid-phase-extraction followed by gas chromatography/mass spectrometry. TBP, TCEP and TBEP were detected at mean values of 622 ng l(-1), 352 ng l(-1), and 2955 ng l(-1), respectively in municipal waste water effluents. This points to a major input of these compounds into the Oder River by municipal waste water discharge. The concentrations of TBP and TBEP decreased downstream the Oder River possibly due to aerobic degradation. TBP, TCEP and TBEP were detected in ground water influenced predominantly by bank-filtered water. This demonstrates a transport of organic compounds by river water infiltration to ground water. TBP, TCEP and TBEP were also detected in rain water precipitation, roof runoff and ground water predominantly influenced by rain water infiltration. This hints to an input of these compounds to ground water by dry and wet deposition after atmospheric transport. Organophosphate esters were also detected in parts of the aquifer at 21 m depth. This demonstrates low anaerobic degradation rates of TBP, TCEP and TBEP. PMID- 12729281 TI - Automatic continuous river monitoring of nitrate using a novel ion-selective electrode. AB - On-site continuous measurement of nitrate in the River Taw situated in Devon (UK) was performed automatically for two months using a recently developed ion selective electrode (ISE). A non-submersible electrode support system was developed and a field instrument was constructed comprising the nitrate-ISEs, a reference electrode and a temperature probe connected through a pre-amplifier to a data-logger and battery supply. The nitrate-ISE was constructed using a commercially available electrode body with a membrane incorporated into the tip. This rubbery membrane contained the sensor molecule (N,N,N-triallylleucine betaine chloride, 6.5% m/m) covalently bound to polystyrene-block-polybutadiene block-polystyrene, SBS, (43.5% m/m), with 2-nitrophenyloctyl ether (2-NPOE) as mediator (40% m/m). The Nernstian slope was -59.1 mV per decade over a linear range of 1400-0.07 mg L(-1) nitrate-N. The limit of detection was 0.007 mg L(-1) nitrate-N with a selectivity coefficient for nitrate against chloride (k(pot)NO3 , Cl-) of 3.4 x 10(-3). The speed of response was less than a minute over the linear Nernstian range. The lifetime in the laboratory exceeded 5 months with no potentiometric drift over the linear Nernstian range. A temperature correction algorithm was also used to accomodate the temperature changes encountered in the river. The river nitrate results obtained with the ISEs at hourly intervals compared very favourably (R2 = 0.9) with those obtained with laboratory automated chemical determinations made on contemporaneous river samples over a concentration range 0.55-2.00 mg L(-1) nitrate-N. The ISEs did not require re calibration and no deterioration in performance or fouling of the membrane surface was observed during 40 days. PMID- 12729282 TI - Release studies of benzalkonium chloride from hydrogel in a freshwater environment. AB - The use of underwater optical sensors to monitor pollution and climate change processes has led to the development of robust instruments able to be deployed in lakes and seas for months at a time. However, despite this improvement in their durability they are subject to biofouling on their optical ports resulting in erroneous readings. The use of hydrogel coatings containing the cationic surfactant benzalkonium chloride (BAC) has been shown to prevent the development of biofouling for up to 12 weeks in the marine environment. In this study the use of hydrogel coatings in the freshwater environment was less successful with fouling visible at 2 weeks. In both field and laboratory studies a rapid initial loss of BAC from the hydrogel film was observed. The loss is a combination of diffusive and mass flow but the period from 12 to 50 h appeared to fit to diffusion kinetics and a diffusion coefficient of 7.3 x 10(-8) cm2 s(-1) (13 degrees C) was calculated, an order of 10 times greater than that found in seawater. Subsequently the rate of loss of the residual BAC, for which a diffusion coefficient of 5.7 x 10(-10) cm2 s(-1) (15 degrees C) was measured, was too low to prevent the early stages of biofouling. PMID- 12729283 TI - Detection and extraction of endosulfan by metal nanoparticles. AB - One of the most common pesticides in the developing world, endosulfan, can be detected in ppm levels using gold nanoparticles. Endosulfan adsorbs on the nanoparticle surface and upon interaction for a long time, the nanoparticles precipitate from the solution. Interaction with silver is weak, yet adsorption occurs leading to removal of endosulfan from the solution. A multilayer assembly of gold nanoparticles prepared on a glass substrate shows excellent spectrophotometric response suggesting potential applications. PMID- 12729284 TI - New thiol adsorbent grafted on silica gel: synthesis, characterization and employment for heavy metal adsorptions. AB - Silica gel surface has been modified in two reaction steps: (i) the silylating agent 3-mercaptopropyltrimetoxysilane was firstly immobilized to give a surface Sil-SH and (ii) this precursor incorporated an ethylene sulfide molecule to obtain the surface denoted Sil-SSH. This material was characterized by elemental analysis, IR spectroscopy, thermogravimetry, solid state 13C and 29Si NMR, and surface area measurement. These materials were employed as adsorbents for divalent heavy cations from aqueous solutions at room temperature and the isotherms were adjusted to a modified Langmuir equation. The maxima number of moles adsorbed were 1.0, 1.5, 1.6, 2.2, 2.4 and 3.3 mmol g(-1) for Co, Cu, Ni, Cd, Pb, and Hg, respectively. PMID- 12729285 TI - Meningococcal disease. PMID- 12729286 TI - Unrelated umbilical cord blood transplant for beta-thalassemia major. AB - A 5-year-old boy with beta-thalassemia major received an unrelated umbilical cord blood transplantation (URD-UCBT). The URD-UCB was six antigen HLA matched. The infused cell dose was 7.5 x 10(7)/kg nucleated cells. Conditioning included busulfan 20 mg/kg, cyclophosphamide 200 mg/kg, fludarabine 150 mg/kg, thiotepa 6 mg/kg, and antithymocyte globulin 90 mg/kg. The post transplant complications were mild hepatic veno-occlusive disease, acute GVHD grade III, and CMV interstitial pneumonia. The subject has been ex-thalassemic for more than 20 months post transplant. The chronic GVHD was limited and could be controlled by methylprednisolone combined with mycophenolate. This is the first successful report of an unrelated umbilical cord blood transplantation for beta-thalassemia major from China. PMID- 12729287 TI - Kernicterus and G6PD deficiency--a case series from Oman. AB - The relationship between glucose-6 phosphate dehydrogenase (G6PD) deficiency and jaundice in the newborn period is well recognized. However, there is concern about the increasing incidence of kernicterus being reported worldwide, especially due to unrecognized G6PD deficiency and early discharge from hospital after birth. We report a case series of kernicterus from a set-up where the high prevalence rate of G6PD deficiency is known. Fourteen cases of kernicterus were seen during a time period of 6 years, 71 per cent of them had G6PD deficiency. Recent literature is reviewed and possible preventive measures in the light of current information and practices are suggested. PMID- 12729289 TI - The effect of iron therapy on the growth of iron-replete and iron-deplete children. AB - This prospective, double-blind, placebo-controlled trial was designed to study the effect of iron therapy on the growth of iron-replete and iron-deficient children, and to study the change in iron status in iron-deficient children with iron therapy. One hundred and fifty children (aged 6-24 months) were included in the study. After an informed written consent, 100 healthy children, who were iron replete (group I) according to preset criteria, were randomly allocated to receive iron supplements 2 ng/kg/day (group IA) or placebo (group IB). Fifty iron deficient children (group II) were administered iron syrup 6 mg/kg/day. Growth parameters (weight, length and head-circumference) and hematological parameters were studied for 4 months. Iron therapy, as compared with placebo, produced a significant improvement of mean monthly weight gain (p < 0.001) and linear growth (p < 0.001) in the iron-deficient children. However, it significantly decreased the weight gain (p < 0.001) and linear growth (p < 0.001) of iron-replete children. Caution should therefore be exercised while supplementing iron to children with apparently normal growth and when the iron status of the child is not known. PMID- 12729288 TI - The impact of HIV infection and trimethoprim-sulphamethoxazole prophylaxis on bacterial isolates from children with community-acquired pneumonia in South Africa. AB - The aim of this study was to compare the type and antimicrobial resistance patterns of bacteria cultured from blood or respiratory tract secretions by HIV status and the use of trimethoprim-sulphamethoxazole (TMP-SMX) prophylaxis in children hospitalized with community-acquired pneumonia. During a 1-year prospective study in Cape Town, South Africa, 250 children [median aged 6 (3-16) months] hospitalized with pneumonia were enrolled; 151 (60.4 per cent) were HIV infected. The incidence of bacteremia [35 of 244 cultures (14.3 per cent)] did not differ by HIV status. Bacteria were cultured in 17 of 32 (53 per cent) bronchoalveolar lavage specimens (BAL), 128 of 210 (61 per cent) induced sputa and 166 of 231 (71 per cent) nasopharyngeal specimens (NPAs). The type and number of bacteria in respiratory secretions did not differ by HIV status, except for a higher rate of Staphylococcus aureus in sputum or BAL [22 of 146 (15 per cent) vs. 3 of 96 (3 per cent), p = 0.003] and NPAs [41 of 135 (30 per cent) vs. 9 of 96 (9 per cent), p < 0.001] of HIV-positive children. The use of TMP-SMX prophylaxis in HIV-infected children was associated with an increased nasopharyngeal carriage of S. aureus [22 of 51 (43 per cent) vs. 17 of 79 (22 per cent), p = 0.009]. The rising prevalence of HIV infection and the use of TMP-SMX prophylaxis may alter the spectrum of colonizing and pathogenic bacteria in children in developing countries. PMID- 12729290 TI - A study of feeding patterns in young infants. AB - With the aim of studying the feeding patterns in infants under 6 months of age, 451 mothers attending the children's clinic in two university hospitals in the north of Tehran were interviewed. All babies had been born in hospital and > 98 per cent had been breastfed during the first few hours of birth. The rate of full breastfeeding at 6 months of age, with no introduction of the bottle, was 83 per cent; approximately 6.5 per cent of infants were fed on breast and bottle concomitantly, and in about 10.5 per cent breastfeeding had been discontinued before 6 months and the babies were fed on bottle only. About 60 per cent of mothers who stopped breastfeeding, did so during the first postnatal month and another 20 per cent during the 2nd and 3rd month after the babies' birth. The mother's age, education or parity, did not affect the rate of breastfeeding. Low birthweight, especially birthweight less than 2 kg, was a risk factor for early termination of breastfeeding. Caesarean delivery and hospitalization of the infant during the neonatal period was also associated with a higher rate of bottlefeeding compared with newborns who had been delivered normally, discharged early, and nursed at home. Although breastfeeding rates are high, the finding that the majority of mothers who give up breastfeeding do so in the early weeks, calls for better support to all mothers by committed health personnel during the period when breastfeeding is being established, and for extra assistance to women whose infants are hospitalized or have a low birthweight. PMID- 12729291 TI - Diarrhea in the Dominican Republic: determinants of the utilization of children's health services. AB - Diarrhea is a leading cause of pediatric morbidity and mortality in the Dominican Republic. A potential policy option for reducing the burden of this disease would be to utilize the public and private sector health resources of the country in a co-ordinated fashion. To do so requires an understanding of the determinants of health services' utilization for this disease. This study examines the predictors of (a) health services' utilization, and (b) public vs. private sector use, for diarrhea in the under-five population in the Dominican Republic. The DHS-2 dataset (1991) was utilized for analysis. Logistic regression models for predicting use and non-use, and for predicting private vs. public sector use were constructed. Age, nutritional status, rural location, higher maternal education and duration of illness greater than 2 days were significant predictors of use of health services. Nutritional status, private insurance and higher maternal education were found to be significant predictors of private sector utilization rather than public sector utilization. Such disease-specific sector utilization should be kept in mind when attempts are made to adjust the public/private mix in health sector reform. PMID- 12729293 TI - Prevalence of exercise-induced bronchospasm in schoolchildren: an urban-rural comparison. AB - To study the prevalence of exercise-induced bronchospasm (EIB), 400 schoolchildren, aged 7-15 years, 200 each from urban and rural areas were administered a standard respiratory questionnaire, spirometry and exercise challenge. The study documented a higher prevalence of EIB in urban areas compared with rural area, 13 vs. 10 per cent. The family members of urban children had significant histories of asthma and allergy. PMID- 12729292 TI - Hypo- and hypermagnesemia in an Indian Pediatric Intensive Care Unit. AB - Data on magnesium disturbances in critically ill children admitted to a Paediatric Intensive Care Unit (PICU) are scarce, especially from developing countries. We have studied occurrence and incidence of hypo- and hypermagnesaemia in children admitted to a PICU and the correlation between such disturbances and the outcome of illness. A total of 100 children (68 boys, 32 girls) aged 6 months to 12 years (mean +/- SD 4.9 + 3.5 years) admitted consecutively to a PICU were studied. At admission and on every alternate day venous blood was obtained for the estimation of serum and RBC-magnesium, serum calcium, sodium, and potassium, and arterial blood for ionized calcium and pH. This was done after ethical approval and informed consent. Hypomagnesaemia and hypermagnesaemia occurred in 60 per cent and 4 per cent of patients, respectively. The incidence of hypomagnesaemia was 30.1, and hypermagnesaemia was two episodes per 100 patient days. The incidence of low RBC-Mg was 17.3 episodes per 100 patient days. Hypomagnesaemia was most common in patients with raised intracranial pressure (63 episodes per 100 patients days). Mortality was nine-fold higher in hypomagnesaemic (30 per cent, 19 of 63) compared with normomagnaesemic (3.3 per cent, one of 30) patients. If Mg and Ca both were low, the mortality rate was 33 per cent (15 of 45 patients) in contrast to nil if both were normal (p < 0.05). We conclude that hypomagnesaemia and low RBC-Mg are a common occurrence in PICU patients and are associated with higher mortality. PMID- 12729294 TI - Evaluation of the outcome of patients admitted to the pediatric intensive care unit in Alexandria using the pediatric risk of mortality (PRISM) score. AB - The aim of this prospective study was to evaluate the use of pediatric risk of mortality (PRISM) score to predict the patient outcome in Alexandria Pediatric Intensive Care Unit (PICU). The study included all admissions to a tertiary care teaching hospital for 13 months. All patients were subjected to thorough history taking and clinical examination. The PRISM score was obtained within 8 h from admission (including 14 parameters with 34 variables). The primary affected system, referral site, number of organ failure on admission, length of hospital stay (LOS) and outcome of patients were recorded. The bed occupancy rate, turnover rate, average LOS, total and adjusted death rates were also recorded. Results showed that the total and adjusted mortality rates were 50 and 38 per cent respectively (n = 205/406 and 125/326, respectively). The mean PRISM score on admission was 26. Non-survivors showed a significantly higher mean score compared with survivors (36 vs. 17). Non-survivors compared with survivors, were significantly younger (12 vs. 23 months), had shorter LOS (3.8 vs. 5.3 days), three or four organ system failure on admission (77 vs. 25 per cent, and 9 vs. 0 per cent of patients) and had significantly higher percentage of sepsis syndrome and neurological diseases, as the primary affected system (20 vs. 10 per cent and 26 vs. 16 per cent). The PRISM score showed a significant positive correlation only with the number of organ failure on admission (r = 0.8104; p < 0.001). The cut-off point of survival was a PRISM score 26 with expected/observed ratio of 1.05 for non-survivors with 91.6 per cent accuracy. Multiple logistic regression analysis revealed that PRISM score, LOS, and the primary affected system were relevant predictors of patient outcome in PICU. In conclusion, the PRISM score is proved to be a good predictor of outcome for children admitted to a PICU with a cut-off point of 26. The mortality in the PICU is affected by LOS, primary system affected, and number of organ failure on admission. PMID- 12729295 TI - Hypothermia on admission: a risk factor for death in newborns referred to the Pernambuco Institute of Mother and Child Health. AB - The objective of this study was to determine the mortality risk related to hypothermia at the moment of admission and other factors such as clinical and geographical related to the transportation of the newborns admitted to the Instituto Materno Infantil de Pernambuco IMIP from 8 March to 11 June 2000. A prospective study involving 320 newborns arriving from home or health centres was carried out. Babies that were dead on arrival or subsequently transferred to other units were excluded. The risk of death was determined according to exposure to hypothermia and other types of exposure, using logistical regression. The risk of death was higher (RR = 3.09; CI = 2.15-4.43) in the group exposed to moderate hypothermia (temperature between 32.5 degrees C and 35.99 degrees C) than in the non-exposed group (temperature equal to or greater than 36.00 degrees C). The relative risk of death was also higher for newborns with a weight of less than 2500 g, that were less than 1 day old, respiratory distress syndrome, premature babies or with congenital malformations, that had used oxygen and/or intravenous infusion during transit, that came from the interior and that had travelled more than 150 km. In the final result of the multivariate analysis, sepsis ('adjusted' RO = 6.23; 95% CI = 5.66-6.80), respiratory distress syndrome ('adjusted' RO = 5.28; 95% CI = 5.03-5.59), moderate hypothermia ('adjusted' RO = 3.49, 95% CI = 3.18-3.81), and distance undertaken greater than 50 km ('adjusted' RO = 2.39; 95% CI = 2.14-2.63) remained. Hypothermia on admission showed itself to be an important and independent risk factor for neonatal death. PMID- 12729296 TI - Blood lead levels and risk factors for lead toxicity in children from schools and an urban slum in Delhi. AB - This cross-sectional study was conducted to estimate the mean blood lead levels (BLL) and prevalence of lead toxicity in a representative sample of schoolchildren and children residing in an urban slum. In addition, the association of potential environmental risk factors with elevated BLL was studied. Children aged 4-6 years were selected from schools of the South zone of Delhi (n = 125) and from an urban slum (n = 65). Risk factors were recorded using a pre-tested questionnaire and blood lead and zinc protoporphyrin (ZPP) levels were estimated. The mean BLL was 7.8 microg/dl (SD 3.9) and the proportion of children with blood lead > or = 10 microg/dl was 18.4 per cent. Distance of the residence or school from a main road appeared to be associated with higher blood lead concentrations, but these differences were not statistically significant. In our setting, vehicular pollution may be a major contributing factor in lead contamination of the environment. PMID- 12729297 TI - Cryptococcal meningitis in pediatric AIDS. AB - Cryptococcus neoformans is an infrequent cause of meningitis in children. We report two cases of HIV positive children of HIV non-reactive parents who were diagnosed as suffering from cryptococcal meningitis. Treatment with amphotericin B and flucytosine was instituted in both the children who recovered and are doing well. PMID- 12729298 TI - Congenital rickets. AB - Although rickets in premature newborns is known to occur, term babies presenting at birth is uncommon. We report a term baby born to a mother with osteomalacia, and presented at birth with signs of florid rickets which was confirmed biochemically. After 4 weeks of treatment, radiological signs of healing were seen. PMID- 12729299 TI - Eradication of Cryptosporidium in four children with acute lymphoblastic leukemia. AB - Four children on chemotherapy for acute lymphoblastic leukemia presented with severe diarrhea and dehydration. Cryptosporidium was identified in the stools using modified Ziehl-Neelsen stain. Two of them received paromomycin and responded well. One was started on paromomycin for 10 days and although there was clinical improvement, his stools examination continued to be positive for Cryptosporidium. He then received azithromycin for 10 days. He responded well and his stools became negative for Cryptosporidium. The fourth patient received azithromycin from the start and responded well. Cryptosporidium should be considered in all immunocompromised children with severe or prolonged diarrhea, and since it is not seen in a routine ova and parasite examination, the laboratory should be notified for diagnostic confirmation using modified Ziehl Neelsen stain. Immunocompromised children with Cryptosporidium diarrhea may benefit from paromomycin or azithromycin therapy. PMID- 12729300 TI - Building research culture. PMID- 12729301 TI - Credit where credit is due. PMID- 12729302 TI - Opposed to unopposed. PMID- 12729303 TI - The family farm. PMID- 12729304 TI - Teaching evidence-based medicine in a small rural family practice office. PMID- 12729306 TI - Requiem for a resident. PMID- 12729305 TI - The University of Missouri Rural Obstetric Network: creating rural obstetric training sites for a university-based residency program. AB - This paper describes a rural obstetric experience that was developed for a university-based family practice residency program and designed to increase the number of deliveries per resident, the number of graduates practicing in rural areas, and the number of graduates doing obstetrics. Rural hospitals can be a source of deliveries for residency training programs. This rural obstetric experience also offers more training months in a rural setting and more months training with family physicians. PMID- 12729307 TI - Service above self. PMID- 12729309 TI - Real-time information-seeking behavior of residency physicians. AB - BACKGROUND: Practicing physicians rarely apply evidence-based medicine (EBM). Unlike their more-experienced counterparts, current residents probably participated in EBM curricula during medical school. The current study was designed to determine the extent to which they or their faculty spend time searching for evidence-based answers. This information will help guide the content and format of EBM curricula in residency programs. METHODS: The method used was direct observation of 13 faculty and 25 residents combined with self report. Number of clinical questions, sources consulted, search times, and satisfaction with answers were collected for clinical encounters. The setting was the University of California San Francisco-Fresno Family Practice Residency Program. RESULTS: Participants asked 274 clinical questions over 215 patient encounters (1.3 per encounter). Residents generated 1.5 per encounter and faculty generated .8. The group sought immediate answers to 66% of questions, found satisfactory answers to 87% of these, and later pursued answers to only 6% of remaining questions. Most searches (66%) took less than 2 minutes. Physicians most commonly used another person or a pocket reference. CONCLUSIONS: Residency physicians have clinical questions but rarely use evidence-based sources to answer them. PMID- 12729310 TI - Point-of-care information that changes practice: it's closer than we think. PMID- 12729308 TI - Use of on-line evidence-based resources at the point of care. AB - BACKGROUND AND OBJECTIVES: The utility of on-line evidence-based summary databases for answering clinical questions at the point of care is not well understood. Our objectives were to determine if family physician faculty could answer their questions using on-line resources and the proportion of answers that influenced patient care. METHODS: This was a prospective study in which clinical faculty in an urban residency training office recorded their clinical questions and their search results. RESULTS: Faculty asked 92 questions. Therapy, prognosis, and epidemiology questions were the most common types of inquiries. Fifty-four percent of the questions were fully or partially answered by use of an on-line resource; obtaining an answer required 5-10 minutes of searching. Physicians reported that 62% of the obtained answers modified their opinion, influenced the care of the current patient 56% of the time, and would affect the care of future patients 70% of the time. Slow Internet connection and interruptions were the most frequent barriers reported. DISCUSSION: Practicing physicians inexperienced in the use of on-line evidence-based resources answered a proportion of their clinical questions that was comparable to reports of more experienced searchers; however, the time required to find answers limits the practical use of these databases during patient care time. On-line summary databases such as those used in this study show promise in providing answers that influence care during the patient's visit. With faster Internet connection (or handheld devices) and improved navigability, such resources have the potential to optimize health care in the primary care setting. PMID- 12729311 TI - Documenting procedures and deliveries during family practice residency: a survey of graduates' experiences, preferences, and recommendations. AB - BACKGROUND AND OBJECTIVES: The University of Washington Family Practice Residency Network (UW Network) is in the process of implementing a Palm Pilot-based procedure and delivery documentation system throughout 16 residency programs. Our study examined the experiences of past UW Network graduates in obtaining hospital privileges and in documenting procedures and deliveries. METHODS: A survey was mailed to 201 1999 and 2000 UW Network graduates, asking them questions about their experiences obtaining hospital privileges after graduation and documenting procedures and deliveries during their training. RESULTS: A total of 124 surveys (62% response rate) were analyzed. Ninety-four percent of the respondents had applied for hospital privileges, and 84% received all the privileges they requested. Forty-four percent indicated they had to provide some written documentation to get hospital privileges, but only 7% had to provide more than a numeric total of procedures or deliveries. Respondents predominantly used log cards and Palm Pilots for data collection. Palm Pilots were preferred over log cards, and the Palm Pilot systems received higher satisfaction ratings. CONCLUSIONS: For the majority of graduates, detailed delivery and procedure information was not necessary to obtain hospital privileges. Nevertheless, there are other reasons to document training experiences, and graduates strongly advise family practice residents to record their procedure and delivery experiences. Family practice residency programs should consider giving house staff handheld computers to record the procedures they perform. PMID- 12729312 TI - The current state of esophagogastroduodenoscopy training in family practice residency programs. AB - BACKGROUND AND OBJECTIVES: Esophagogastroduodenoscopy (EGD) is a useful diagnostic procedure to evaluate patients with upper gastrointestinal complaints. Although family physicians have demonstrated that they can competently perform EGD, only a minority of family physicians perform EGD. This study determined the current state of EGD training in US family practice residency programs and how often graduating residents seek EGD privileges. METHODS: We conducted a cross sectional descriptive study surveying program directors from all Accreditation Council for Graduate Medical Education-approved family practice residency programs regarding EGD training in their program. RESULTS: Of the 471 surveys mailed, 441 (94%) were returned. A total of 143 (32%) program directors reported that their program offered EGD training, but only 58 (13%) actually trained at least one resident. Residents performed a mean of 20 +/- 2.4 EGDs per resident, and residents trained by family physicians performed more EGDs than residents trained by other specialties. In July 2000, .04% of graduating family practice residents sought credentials for EGD, and their training occurred in 32 (7%) residency programs. CONCLUSIONS: Only a minority of family practice residents seek credentialing after residency to perform EGD. PMID- 12729313 TI - The effect of medical students' international experiences on attitudes toward serving underserved multicultural populations. AB - BACKGROUND: We evaluated the effect of international electives on the attitudes of preclinical and clinical-year medical students with respect to serving underserved multicultural populations. METHODS: A self-assessment instrument was used to measure attitudes of 146 students before and after participating in international electives. The same attitudinal items were also analyzed at two time intervals for 18 students who completed international electives as preclinical students and 76 class cohorts who did not. RESULTS: Analyses show that the effect of international experiences is different for preclinical students and clinical students. For both groups, however, these experiences can develop and support perceptions and values conducive to serving underserved multicultural populations. These include reported increases in cultural competence and important personal attributes like idealism and enthusiasm. In addition, these experiences can heighten clarity about career roles, including those involving underserved multicultural patients. CONCLUSIONS: This study provides support for the hypothesis that international electives develop attributes that could benefit underserved multicultural populations. PMID- 12729314 TI - Family narratives, culture, and patient-centered medicine. AB - BACKGROUND AND OBJECTIVES: As part of our family medicine clerkship seminar on the patient-physician relationship, third-year students write about an illness episode within their own families. METHODS: Using a grounded research approach, we examined 260 student narratives to extract the most significant meanings. RESULTS: Significant themes that emerged include the role of family members in illness episodes, specific influences resulting from the family's ethnicity or religion, experiences with socially unacceptable illnesses, experiences with death, appreciation of the moral trajectory of illness, and situations that display the fallibility and limitations of medicine. CONCLUSIONS: Writing exercises can help students recognize the centrality of narrative and of cultural values in medicine so they are better able to understand their patients and provide more patient-centered medical care. PMID- 12729315 TI - Clinical research in family medicine: quantity and quality of published articles. AB - BACKGROUND AND OBJECTIVES: Publication of clinical research in peer-reviewed journals is an important measure of scholarly productivity. This study determined the quantity and quality of original clinical research published by family physicians. METHODS: We surveyed clinical research papers published in the year 2000 in four leading family medicine research journals and research originating in a family practice institution but published in 16 non-family medicine journals. All were selected on the basis of relevance to family physicians and "impact factor." The relevance and validity of papers was assessed using previously established criteria. RESULTS: The survey of family medicine journals revealed a total of 170 original research articles. Ninety eight were from academic family practice programs, and the remaining 72 were from other medical specialties or health care institutions. Most of the papers were cross-sectional surveys. There were seven qualitative studies, six randomized controlled trials, and no systematic reviews from family practice programs in these journals. Eight of the articles were from practice-based research networks. A total of 79 articles were considered relevant or highly relevant, and 22 of these were also considered valid (Patient-oriented Evidence That Matters or POEMs). The survey of 16 non-family medicine journals revealed 37 clinical research papers: 16 surveys, nine prospective cohort studies, seven randomized controlled trials, three systematic reviews/meta-analysis, one qualitative study, and one case-control study. There were nine "highly relevant" papers--seven could be classified as POEMs. CONCLUSIONS: Most clinical family medicine research uses less-rigorous study designs, such as the cross-sectional survey. The majority of papers do not meet established criteria for relevance and validity. There are no standards or comparable studies to compare these results to prior years or to other disciplines. PMID- 12729316 TI - Reframing Balint: thoughts on family medicine departmental Balint groups. AB - This paper explores recurrent processes and themes in the 1,300 family medicine faculty, resident, intern, and community Balint groups the author has facilitated/led. The frequent group "deviation" from the central Balint task of understanding difficult physician-patient relationships is reframed as less "resistance" or "obstacle" to work, as it is an expression of unmet developmental needs and organizational realities. When group members are carefully attended to (by facilitator and one another), the group often becomes emotionally capable of addressing a "case " in the conventional Balint understanding of the work the group has assembled to do. The group dynamics of such "hybrid" Balint groups thus become comprehensible as other than error. PMID- 12729317 TI - Family medicine and research: from here to eternity. PMID- 12729318 TI - When should immunosuppressants be prescribed to treat systemic vasculitides? AB - Steroids and immunosuppressants are indicated to treat systemic vasculitides. However, the therapeutic strategy is different from one disease to the other. Treatment choice should be adapted to the predictable outcome, severity, pathogenic mechanisms and patient's general condition. In polyarteritis nodosa, Churg Strauss syndrome, and microscopic polyangiitis we have demonstrated that immunosuppressants should not be systematically prescribed. Immunosuppressants should be only prescribed in the most severe patients, when factors of poor prognosis are present. In Wegener's granulomatosis, immunosuppressants should be systematically prescribed together with steroids. The optimal treatment duration is of 12 months for polyarteritis nodosa and Churg-Strauss syndrome. A more prolonged treatment is mandatory in Wegener's granulomatosis, at least 18 months. The new therapeutic strategies comprise also new immunosuppressants and new immunomodulating agents which could replace or be associated to the "older drugs". PMID- 12729319 TI - Metronidazole plus ciprofloxacin therapy for active Crohn's disease. AB - OBJECTIVE: The aim of this study was to investigate the efficacy of metronidazole plus ciprofloxacin for the treatment of Japanese patients with active Crohn's disease. METHODS AND PATIENTS: Seven patients (counting one patient twice with 2 enrollments at a 5-month interval) with a flare-up of Crohn's disease were enrolled. While continuing the baseline treatment under which the patients relapsed, they received metronidazole 250 mg twice (4 patients) or three times (3 patients) daily plus ciprofloxacin 200 mg three times daily for 4 weeks. The efficacy was evaluated by the changes in the assessment score of IOIBD, the International Organization for the Study of Inflammatory Bowel Disease, and the inflammation markers: C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and white blood cell count. RESULTS: Metronidazole plus ciprofloxacin decreased the CRP in seven patients and the IOIBD score in six patients. Significant differences were detected in these parameters at weeks 2 and 4 compared with baseline. Five of the patients achieved normalization of CRP and a reduction of the IOIBD score to zero or one. Although one patient complained of taste disturbance, no other adverse events occurred and all patients completed the 4 weeks of study medication. CONCLUSION: The addition of metronidazole plus ciprofloxacin could be a useful intervention for the treatment of Japanese patients with active Crohn's disease. PMID- 12729320 TI - Genomic mutations with amino acid substitutions of circulating hepatitis B virus found in non-B, non-C patients with hepatocellular carcinoma. AB - OBJECTIVE: Most hepatocellular carcinoma (HCC) in Japan is caused by chronic infection with hepatitis B virus (HBV) or hepatitis C virus (HCV). HBV DNA has been detected in the serum and liver tissue of some proportion of those patients who are HBs antigen-negative and HCV antibody-negative; i.e., non-B, non-C (NBNC) patients with HCC. We sought to detect HBV DNA in the serum from NBNC HCC cases and to investigate genomic mutations of HBV in seronegative cases. PATIENTS AND METHODS: The sera from 26 NBNC HCC patients were examined by polymerase chain reaction (PCR) followed by southern blotting for existence of HBV DNA. The precore/core and polymerase regions of the HBV genome in the sera from five seronegative cases were analyzed by direct sequence. RESULTS: HBV DNA was detected in 17 of 26 patients (65.4%). Demographic factors such as age, gender, anti-HBs positivity, anti-HBc positivity, complication with cirrhosis, and excessive alcohol intake did not affect circulating HBV positivity. Genomic mutations with amino acid substitutions were detected in the polymerase and the precore regions from one of the five cases, and in the core region from four of the five cases. CONCLUSIONS: PCR-based HBV screening is necessary in patients suffering from liver diseases of unknown etiology, although its etiological importance and benefit of viral elimination have not been established. Genomic mutations in the precore/core and the polymerase region detected in this study might be involved in the lack of HBsAg in NBNC HCC cases. PMID- 12729321 TI - Autoimmune hepatitis associated with Graves' disease. AB - A 31-year-old woman with Graves' disease with a 12-month-history of propylthiouracil intake and autoantibodies in the sera was admitted to our hospital. The differential diagnosis between autoimmune hepatitis and propylthiouracil-induced hepatitis was intractable. Steroid therapy was started and she showed a complete response to the treatment. Liver biopsy demonstrated acute hepatitis and plasma cell infiltration. A second liver biopsy, which was performed 10 months after starting steroid therapy, showed some inflammatory cells in the portal tracts. These findings suggest that she had been suffering from autoimmune hepatitis. PMID- 12729322 TI - Surgical resection of malignant lymphoma in the right atrium after systemic chemotherapy. AB - A 74-year-old man was referred to us for evaluation of a tumor in the right atrium (RA). Transesophageal echocardiography (TEE) showed an unmovable 50x60 mm mass in the RA. Based on histological findings of subcutaneous tumors in the right abdominal wall, he was diagnosed as malignant lymphoma (ML), and treated with a THP-COP regimen. Upon completion of first THO-COP therapy, TEE showed marked regression of the mass and division into 3 masses, one of which showed marked floating movement with a small stalk. To prevent the risk of embolic events, surgical resection was performed. Resected tumors were necrotic tissues. Serial imaging of cardiac tumor and surgical resection is desirable to decrease the possibility of embolic complication. PMID- 12729323 TI - Hypercalcemia due to vitamin D intoxication with clinical features mimicking acute myocardial infarction. AB - We report a case of hypercalcemia in an elderly patient due to vitamin D intoxication with clinical features and electrocardiogram (ECG) findings mimicking acute myocardial infarction. A 78-year-old man was referred to our department with symptoms of general fatigue, anorexia and chest pain. The ECG demonstrated ST elevation in leads V1 to V3 and diffuse T wave flattening, resulting in myocardial infarction being suspected. However, his symptoms, including chest pain, gradually improved and the ECG returned to normal in accordance with a fall in his serum calcium level. We introduce the use of QaTc interval shortening in differentiating ST-T changes of hypercalcemia from those of true myocardial ischemia. PMID- 12729324 TI - Chinese herbs and bone disease. AB - We treated a patient with an unusual bone disease at least partly associated with Chinese herbs. Seven years after 65-year-old man had begun to consume Chinese herbs, multifocal osteoarthralgias were noted, and the patient was hospitalized for renal dysfunction (serum creatinine, 2.8 mg/dl; urea nitrogen, 19 mg/dl). Fanconi syndrome also was apparent. A renal biopsy specimen showed tubulo interstitial fibrosis. Chinese herbs were discontinued and prednisolone was started, but bone and joint pain as well as renal function gradually worsened. Four years later, creatinine was 9.0 mg/dl and alkaline phosphatase was 571 IU/l. As bone scintigraphy revealed localized asymmetric lesions, Paget's disease of bone was suspected at first. However, neither osteosclerosis nor hypertrophy was seen in radiographs. Based on a bone specimen histology we diagnosed as mixed type renal osteodystrophy including osteomalacia and osteitis fibrosa. Mosaic pattern of cement lines was not present. This case was not compatible with either Paget's disease or typical renal osteodystrophy as seen in dialysis patients. Etidronate disodium was effective in alleviating bone symptoms. The patient's bone disorder may be a new disease at least partly related to Chinese herbs independently of nephropathy. PMID- 12729325 TI - Primary effusion lymphoma of the left scrotum. AB - A 51-year-old man without human immunodeficiency virus, hepatitis B virus or hepatitis C virus was admitted with left scrotum swelling and hydrocele. The cytological finding of fluid in the left scrotum revealed malignant lymphoma, and the immunophenotypic analysis and monoclonal rearrangement of immunoglobulin heavy chain demonstrated B-cell lymphoma. However, no solid tumor of lymphoma was identified in the specimen following a left orchiectomy, or in any other body site and genomic human herpes virus-8 and Epstein-Barr virus were not detected in the lymphoma cells. So we interpreted this as a primary effusion lymphoma without any ethological viral infection. Subsequently, he underwent chemo-radiation therapy and has remained in remission. PMID- 12729326 TI - Primary cutaneous diffuse large B cell lymphoma: a clinically aggressive case. AB - An 80-year-old woman was diagnosed with primary cutaneous B cell lymphoma. She had multifocal cutaneous tumors, but no nodal lymphadenopathy or bone marrow involvement. Histopathological examination of a biopsy specimen showed diffuse large B cell lymphoma without a bcl-2 expression. Complete resolution of hypercalcemia and disappearance of tumors were achieved with CHOP therapy, but a rapidly progressive skin lesion was observed soon after the completion of the therapy. The clinical manifestation of primary cutaneous diffuse large B cell lymphoma is diverse and the treatment strategy is not entirely clarified. This case represents another example of this rare lymphoma. PMID- 12729327 TI - Primary antiphospholipid syndrome: a cause of fever of unknown origin. AB - Antiphospholipid syndrome (APS) is defined as the occurrence of thrombosis, recurrent miscarriage, or both in association with laboratory evidence of persistent antiphospholipid antibodies. Owing to protean manifestations and laboratory studies, the diagnosis may be difficult. Because the other signs and symptoms of thrombosis are predominant, prolonged fever is not usually the main clinical finding. We describe a patient who presented with fever of unknown origin (FUO) and was found to have thromboses of the splenic vein, the superior mesenteric vein, and the portal vein due to the primary antiphospholipid syndrome. We also reviewed the medical literature (Medline 1966-2001), including the main FUO series of the previous 40 years, and laparotomy series for FUO. We conclude that although very rare, primary APS and thrombosis may present with FUO. APS should be considered in the differential diagnosis of prolonged fever associated with thrombosis. PMID- 12729328 TI - Cytomegalovirus colitis following immunosuppressive therapy for lupus peritonitis and lupus nephritis. AB - We report a woman with lupus nephritis complicated with lupus peritonitis and cytomegalovirus (CMV) colitis. Diagnosis of lupus peritonitis was made by abdominal computed tomography scan, colonoscopy, and ascitic fluid analysis. Steroid and cyclophosphamide therapy resulted in the improvement of severe lupus nephritis and peritonitis. Thereafter, she developed multiple colonic ulcers as diagnosed by colonoscopy and positive CMV antigenemia assay. Treatment with ganciclovir resulted in the disappearance of colonic lesions. The low cluster of differentiation (CD)4+ lymphocyte count (41/mm3) suggested that the cell-mediated immunity of this patient was comparable to that seen in patients with acquired immunodeficiency syndrome (AIDS). PMID- 12729330 TI - Primary gastric plasmacytoma at an early stage: unremarkable effect of eradication of Helicobacter pylori. PMID- 12729329 TI - Norwegian scabies in an elderly patient who died after treatment with gammaBHC. AB - We report the first case of an elderly patient suffering from Norwegian scabies who died after treatment with gammaBHC. GammaBHC (60 mg) was applied over the entire body except for the head of a 91-year-old female in our geriatric ward. This chemical was used in a smaller dose than previously reported, and it proved to be effective except for its effect on the head. Six days after the initial application, gammaBHC (40 mg) was also applied to the patient's head, but she died 5 days later. We propose that application of gammaBHC to the head of such patients should be done with great caution. PMID- 12729331 TI - Urinary bladder infection/irritation among female users of a high-tech toilet. PMID- 12729332 TI - Clostridium subterminale septicemia in a recipient of allogeneic cord blood transplantation. PMID- 12729333 TI - Using qualitative research methods to improve health care for patients with TB and lung disease. PMID- 12729334 TI - Olaf Scheel and Johannes Heimbeck: their contribution to understanding the pathogenesis and prevention of tuberculosis. AB - From 1924 onwards, mandatory tuberculin testing (von Pirquet's method) was introduced for student nurses on entry to the Ullevaal Hospital School of Nursing, Oslo. The chief physician of the hospital's Department IX, Olaf Scheel, was responsible for this measure. In 1927, his deputy Johannes Heimbeck showed that about half of the students were not infected at the time of entry, a conclusion that was in fundamental conflict with the prevailing view that nearly all tuberculous infection occurred in childhood. Virtually all tuberculin negative student nurses, however, became infected during their 3-year training. These findings changed our understanding of the pathogenesis of tuberculosis. BCG vaccination had recently been introduced by Calmette. From 1927 onwards Heimbeck offered BCG vaccination to tuberculin-negative student nurses, while Scheel undertook a similar project among medical students. The two projects continued until respectively 1936 and 1939. Long-term follow-up of both groups demonstrated a protective effect of about 80% in those accepting vaccination. Calmette had given the vaccine per os to new-born babies. Heimbeck and Scheel pioneered giving the vaccine via injection and to adults. These projects have been criticised for being based on voluntary inclusion and not being conducted as randomised control trials. The results were so convincing, however, that they led to the launch of the Norwegian BCG programme shortly after World War II. Scheel and Heimbeck' efforts were also of great importance for the use of the BCG vaccine in other countries. PMID- 12729335 TI - A comparison of dual skin test with mycobacterial antigens and tuberculin skin test alone in estimating prevalence of Mycobacterium tuberculosis infection from population surveys. AB - SETTING: City of Manaus, Amazonas, Brazil. OBJECTIVE: To compare estimates of Mycobacterium tuberculosis infection prevalence obtained using traditional tuberculin skin test (TST) criteria and dual skin test (DST) data. METHODS: A total of 1070 schoolchildren received DST with tuberculin and four environmental mycobacteria sensitins. Responses were classified as sensitin-dominant, tuberculin-dominant or non-dominant. Positive predictive values (PPV) were defined using 'narrow' and 'wide' standards based on DST responses. These predictive values were derived for each category of tuberculin indurations, and were used to calculate the prevalence estimates. RESULTS: Using DST data, the estimates of M. tuberculosis prevalence for scar-negative children were 7.4% (M. avium) and 7.8% (M. scrofulaceum) using the 'narrow' standard, and 16.9% (M. avium) and 15.2% (M. scrofulaceum) using the 'wide' standard. The percentage with TST > or =10 mm was 11.5%. Scar-positive children had higher estimates using both the 10 mm cut-off and DST data. CONCLUSION: In settings with a relatively low prevalence of M. tuberculosis infection and high cross-reactivity with environmental mycobacteria, DST can help to assess the validity of traditional thresholds for estimating the prevalence of M. tuberculosis infection. DST data with environmental antigens and tuberculin do not distinguish BCG-induced cross reactivity. PMID- 12729336 TI - Tuberculin reactivity in adult BCG-vaccinated subjects: a cross-sectional study. AB - INTRODUCTION: Interpretation of the tuberculin skin test (TST) may be complicated by prior bacille Calmette-Guerin (BCG) vaccination. The skin reaction to the vaccination interferes with the management of individuals who may be infected with Mycobacterium tuberculosis. OBJECTIVE: To discriminate between TST reactions due to infection and those due to vaccination in subjects with unknown BCG status. METHODS: Among 60200 subjects tested with 5TU PPD for screening purposes, 4987 contacts of infectious TB cases (Group A), 4962 BCG-vaccinated subjects (Group B) and 5000 subjects from the general population (Group C) were sampled. The frequencies of TST cut-off diameters were calculated for the three groups using a logistic regression model. The frequency of positive subjects in each group and the sensitivity, specificity and predictive values were also computed by means of these cut-offs. RESULTS: The risk of being a contact versus BCG vaccinated increases 2.43-fold with every mm of TST diameter. The 11 mm cut-off point seems to be the best discriminating value. CONCLUSIONS: Using the traditional 10 mm cut-off, we can consider all vaccinated subjects with a positive TST to be infected. The TST remains a valuable tool for the evaluation of household contacts and suspected cases of tuberculosis in BCG-vaccinated subjects and in populations with high vaccination coverage. PMID- 12729337 TI - Non-adherence to tuberculosis treatment in the eastern Tarai of Nepal. AB - SETTING: Tarai districts, eastern Nepal. OBJECTIVES: To identify potential methods of increasing adherence to tuberculosis (TB) treatment by determining factors that patients felt influenced adherence. METHODS: New pulmonary TB patients registered from July to November 1998 with an outcome of non-adherence to treatment (NA) were identified from District TB Registers, traced and interviewed using a semi-structured questionnaire. An equivalent number of adherent (A) patients were interviewed. RESULTS: Of 81 NA patients traced, 30 were interviewed, 16 had been incorrectly classified, age was incorrectly recorded in four, 13 had migrated and 18 were not found. The groups were similar in demographics, type and knowledge of TB. More A patients knew their diagnosis (P = 0.07) and reported haemoptysis as an initial symptom (P = 0.03). NA patients had longer travel to a health facility (P < 0.001), and fewer had been informed by health care workers (HCW) about the consequences of not completing treatment. The most common reasons given for stopping treatment were side-effects, HCWs' mistakes or behaviour, and health service failure. Desire for cure and knowledge that TB was curable were most the important reasons for completing treatment. CONCLUSION: Non-adherence seemed related to treatment delivery failures. The health system needs strengthening in Nepal. Intensified HCW training and supervision, better health education for patients and families, more flexibility for treatment supervisors, adequate supplies for treatment centres and decentralisation of treatment delivery to the lowest health service level practicable are urgently needed. PMID- 12729338 TI - Resistance to anti-tuberculosis drugs and practices in drug susceptibility testing in Moldova, 1995-1999. AB - OBJECTIVE: To evaluate practices in initial drug susceptibility testing (DST) in Moldova, anti-tuberculosis drug resistance and the implications for tuberculosis control. METHODS: Retrospective record review in the national reference laboratory. RESULTS: Of 3463 cases, 57.1% were recorded as 'new' and 24.6% as 'retreatment' cases; previous treatment status was not recorded for 18.3%. Of the 'new' cases, 1655 were correctly classified according to international recommendations and 322 were misclassified. The number of cases increased from 443 in 1995 to 939 in 1999; the proportion of 'retreatment' increased from 17.4% to 35.5%, 'any drug resistance' from 20.3% to 41.6%, and 'multidrug resistance' from 2.7% to 11.2%. In 1998-1999, 'any drug resistance' and 'multidrug resistance' in 800 previously untreated cases were respectively 29.1% and 5.3%, and respectively 61.0% and 21.9% in 521 'retreatment' cases. Of a total of 216 'multidrug-resistant' cases in 1998-1999, 21.8% were reported resistant to ethambutol and 81.5% to streptomycin. CONCLUSIONS: Initial specimens for culture are frequently taken late, after the start of treatment, compromising their usefulness for case management or surveillance. Inadequate treatment has led to an increase in the number of cases, the proportion of previously treated cases and the prevalence of drug resistance. In 1998-1999, a high proportion of cases with 'multidrug resistance' were susceptible to ethambutol. PMID- 12729339 TI - The role of the nurse in the community-based treatment of multidrug-resistant tuberculosis (MDR-TB). AB - SETTING: A community-based treatment program for multidrug-resistant tuberculosis (MDR-TB) in Lima, Peru. OBJECTIVES: To describe the activities carried out by the nurses working with the program. DESIGN: A qualitative study using a variety of ethnographic methods, including participant observation, focus groups, and key informant interviews over a 5-year period. RESULTS: Nurses were responsible for carrying out a wide variety of activities within the program. These included patient-focused activities such as identifying patients, evaluating patients prior to starting and during therapy, and managing emergencies; educational activities for both patients and health professionals managing MDR-TB; and coordination activities, including over-seeing health workers and communicating between team members. CONCLUSION: Nurses play a key role in the community-based management of MDR-TB. PMID- 12729340 TI - Peripheral neuropathy associated with treatment for multidrug-resistant tuberculosis. AB - OBJECTIVE: To review the incidence and management of peripheral neuropathy in patients receiving therapy for MDR-TB. METHODS: A case series with retrospective chart review of 75 patients who initiated individualized therapy for multidrug resistant tuberculosis (MDR-TB) in Lima, Peru, between 1 August 1996 and 31 January 1999. RESULTS: All patients had confirmed MDR-TB and were receiving individualized therapy, comprised of an average of six drugs. Ten (13%) of these patients presented with symptoms of peripheral neuropathy, confirmed by electromyography. All symptoms were reported in the lower extremities, and all were sensory in nature. Median time to presentation from initiation of MDR-TB therapy was 9.1 months. No significant risk factors associated with development of peripheral neuropathy were identified. Management strategies depended on the severity of symptoms and included the treatment of contributing co-morbidities, medications for neuropathic pain, and adjustment of doses of possible offending agents. All patients responded to management; three patients were left with mild residual symptoms. Patients whose neuropathy resolved had symptoms for a median of 7 months. CONCLUSIONS: Peripheral neuropathy was encountered in 13% of our cohort of MDR-TB patients. The diagnosis of peripheral neuropathy can be based on clinical presentation alone, and effective management of this side-effect is possible without sacrificing MDR-TB treatment efficacy. PMID- 12729341 TI - Diabetes modifies the male:female ratio in pulmonary tuberculosis. AB - SETTING: Socio-cultural factors have been invoked to explain the male predominance among patients with pulmonary tuberculosis, but there is no conclusive evidence of their role. OBJECTIVE: To assess male predominance in a group of diabetics with pulmonary tuberculosis compared with patients with pulmonary tuberculosis alone. DESIGN: Clinical records of in-patients with pulmonary tuberculosis and with (TBDM group, n = 202) or without (TB group, n = 226) diabetes mellitus were reviewed, and the male percentages in each of six age groups (15-29, 30-39, 40-49, 50-59, 60-69, > or = 70 years) calculated. RESULTS: In the TB group, no gender difference (51% males) was found in the first age period, followed by a male predominance thereafter (71%, 68%, 75%, 63% and 58%). The TBDM group showed a similar pattern in the first two age groups (56% and 74%), followed by a steadily decline (r(S) = -0.90, P = 0.04) in male percentage (60%, 44%, 45%, 27%), leading to a female predominance after age 50. The association of age and gender was also corroborated by logistic regression in TBDM (P = 0.02), but not in TB (P = 0.19) patients. CONCLUSIONS: Diabetes was associated with a progressive shift of male predominance in pulmonary tuberculosis. Because diabetes is a disease that affects social activities similarly in men and women, our results suggest that factors other than socio cultural ones are also important for determining the male predominance in pulmonary tuberculosis. PMID- 12729342 TI - Granulomatous mammary disease: ten years' experience with fine needle aspiration cytology. AB - OBJECTIVE: To determine the aetiological types of granulomatous disease of the breast in women presenting with mammary complaints in the Sudan. METHODS: Clinical history and physical examination, complete blood counts, Mantoux test, histopathology and fine needle aspiration cytology (FNAC). RESULTS: Granulomatous mastitis was seen in 11/2500 (0.44%) patients with mammary disease over a 10-year period. All were of childbearing age (mean 26.0 +/- 5.9 years). Common presentations were diffuse swelling, well-circumscribed masses, nipple retraction, multiple sinuses and superficial skin ulcers. Lymphadenopathy was seen in more than 60% of the patients. Diagnosis was based on cytomorphological features in 10/11 cases and histopathology in one. Nine were diagnosed with tuberculous mastitis and two with idiopathic granulomatous mastitis. Acid-fast bacilli (AFB) could not be demonstrated in any of the cytology smears. Tuberculous mastitis responded to empirical anti-tuberculosis treatment, with a minimum follow-up of 2 years in seven women. CONCLUSION: Tuberculous mastitis is a rare entity in women with mammary disease in the Sudan. Alternative diagnoses such as idiopathic granulomatous mastitis should be made only after failure of an adequate trial of anti-tuberculosis treatment. FNAC is a useful diagnostic tool even if AFB cannot be demonstrated. PMID- 12729343 TI - NRAMP1 genetic polymorphisms as a risk factor of tuberculous pleurisy. AB - SETTING: Nrampl encoded by the NRAMP1 gene influences the phagolysosomal function of alveolar macrophage against Mycobacterium tuberculosis. Genetic polymorphisms of NRAMP1 affect innate host resistance through the defective production and function of Nrampl. OBJECTIVE: To investigate this relationship, the NRAMP1 polymorphisms in patients with tuberculous pleurisy were determined. DESIGN: Pleural biopsy proven 56 patients were designated to the pleurisy group and 45 healthy adults were designated to the healthy control group. Three NRAMP1 polymorphisms such as single nucleotide change in intron 4(469 + 14G/C, INT4), a non-conservative single-base substitution at codon 543(D543N) and TGTG deletion in the 3' untranslated region (1729 + 55del4, 3'UTR) were determined. RESULTS: The frequencies of mutant genotypes of INT4 and 3'UTR were significantly high in the pleurisy group (P = 0.01, P = 0.02), but the frequencies of D543N were not significantly different between the two groups. Odds ratios (OR), which are a comparison of the wild with the mutant genotype, were 8.02 (95%CI 2.42 approximately 26.57) for INT4 and 5.73 (95%CI 1.14 approximately 28.92) for 3'UTR which were statistically significant. In the combined analysis of the INT4 and 3'UTR, the ORs were 6.00 (95%CI 1.46 approximately 24.64) for GC/++ genotype and 14.00 (95%CI 1.61 approximately 121.75) for GC/+del when compared with GG/++; these differences were statistically significant. CONCLUSION: The NRAMP1 genetic polymorphisms, especially INT4 and 3'UTR, were closely related to tuberculous pleurisy. PMID- 12729344 TI - A comparison of direct microscopy, the concentration method and the Mycobacteria Growth Indicator Tube for the examination of sputum for acid-fast bacilli. AB - SETTING: In countries with high human immunodeficiency virus prevalence, laboratory diagnosis of pulmonary tuberculosis with the standard Ziehl-Neelsen (ZN) technique is characterised by low sensitivity. OBJECTIVE: To compare test characteristics of direct microscopy, the concentration method and the Mycobacteria Growth Indicator Tube (MGIT). DESIGN: Three hundred specimens from patients diagnosed with pulmonary tuberculosis were tested for the presence of mycobacteria. Specimens were stained with ZN, decontaminated by adding 4% NaOH, concentrated by centrifuging and processed in MGIT broth. The gold standard was defined as a positive MGIT culture or a positive acid-fast bacilli smear of material obtained from a negative culture after 42 days. RESULTS: A total of 44 (14.7%) specimens were contaminated. Of 256 valid specimens, 234 (91.4%) were positive according to the gold standard definition. Decontamination and concentration of the sample increased the sensitivity of direct microscopy from 67.5% to 87.1%. Specificity remained unchanged (95.5%). The overall median time to detection of MGIT culture-positive specimens was 5 days, ranging from 4 (direct smear-positive specimens) to 12 days (concentration smear-negative specimens). CONCLUSION: The concentration method substantially increases the sensitivity of direct microscopy without much extra input. The MGIT culture technique has considerable advantages, but its relatively high contamination rate and its high cost make it a less recommendable option for widespread use in routine district laboratories. PMID- 12729346 TI - Surveillance of drug-resistant Mycobacterium tuberculosis in The Gambia. AB - To determine the rates of drug-resistant tuberculosis in The Gambia, Mycobacterium tuberculosis isolates obtained from 225 patients during a nationwide survey were tested against isoniazid, rifampicin, ethambutol and streptomycin using the resistance ratio method. Only nine (4%) of the patients had strains that were resistant to one or more drugs. None of the patients with drug-resistant M. tuberculosis had previously been treated for tuberculosis. Drug resistant tuberculosis is, as yet, not common in The Gambia. Periodic surveys for drug-resistant tuberculosis are recommended to monitor changes that may emerge over time. PMID- 12729345 TI - [The prevalence of respiratory symptomes and immediate hypersensitivity reactions in a population exposed to flour and cereal dust in five flour mills in Morocco]. AB - OBJECTIVE: To evaluate the prevalence of respiratory symptoms, ventilatory function disorder and immediate hypersensitivity reactions in a population exposed to flour and cereal dusts in five flour mills in Morocco. METHODS: The study of 373 exposed and 301 non-exposed subjects consisted of an analysis of working conditions and a medical survey. RESULTS AND CONCLUSION: The prevalence of clinical respiratory symptoms was 64.1% among exposed subjects and 41.2% among non-exposed subjects. Cough, expectoration, rhinitis, conjunctivitis, dermatitis, asthma and chronic bronchitis were more frequent among mill-workers. Spirometry was abnormal in 31.6% of those exposed: anomalies were found in both flow and volume. Among exposed subjects with a ventilatory disorder, 77.9% had only small airways syndrome or a light deficit. Smoking was the cause of excess morbidity. Skin prick tests were positive for at least one allergen in 65.4% of exposed compared to 27.2% non-exposed subjects. The prevalence of positive skin tests to occupational allergens alone was higher among those exposed (42.4%) than among the non-exposed (9.9%). The implementation of adequate medical and technical prevention may reduce this risk. PMID- 12729347 TI - Multidrug-resistant tuberculous meningitis in patients with AIDS. AB - We present clinical manifestations, bacteriologic characteristics, and outcomes for eight patients with multidrug-resistant (MDR) tuberculous meningitis and AIDS. All developed meningitis as a terminal complication of previously diagnosed MDR-TB despite anti-tuberculosis therapy. Seven patients presented with fever, five with headache, four with altered mentation, two with focal deficits and one with seizures. CSF examination revealed pleocytosis, hypoglychorrhachia and elevated protein. Mycobacterium tuberculosis resistant to at least isoniazid and rifampin was isolated from all patients. Intracerebral mass lesions were detected in three patients, hydrocephalus in three, meningeal enhancement in five, and infarcts in two. Seven patients died 1-16 weeks after the diagnosis of meningitis; the eighth was lost to follow-up. MDR tuberculous meningitis is a difficult-to-treat infection with a high fatality rate. PMID- 12729348 TI - Rates of adverse reactions to first and second doses of BCG vaccination: results of a large community trial in Brazilian schoolchildren. AB - OBJECTIVE: To evaluate the incidence of adverse reactions to first and second bacille Calmette-Guerin (BCG) vaccination in schoolchildren. SETTING AND DESIGN: Enhanced surveillance in a Brazilian trial. Suspected reactions were reported to a nurse who visited cases and completed a standard form. RESULTS: Among 71341 schoolchildren studied, 33 reactions were reported. Of these, 25 fulfilled the criteria, resulting in a rate of one per 2854 vaccinations, with no deaths or BCG osis. Reactions to second doses were more common than to first BCG vaccinations, but this difference was not statistically significant. CONCLUSIONS: Adverse reactions to a second dose of BCG may be more frequent than reactions to a first dose, but they are still rare events. PMID- 12729349 TI - The eventual 6th and 7th elements of DOTS: a metaphor. PMID- 12729350 TI - Quality of anti-tuberculosis drugs offered to patients. PMID- 12729351 TI - Medical treatment of multiple pleural hydatid cysts. PMID- 12729352 TI - A two-year regional program for the early detection of cutaneous melanoma. AB - BACKGROUND: A regional program for the early diagnosis of cutaneous melanoma involving general practitioners was effective in 1997-1998 in the Friuli Venezia Giulia region in Northern Italy. The aim of the 2-year program was to evaluate the role of a skin examination performed by general practitioners in people older than 18 years without known skin lesions and spontaneously presenting to their offices for any reason, with referral of suspect cases to a pre-identified regional dermatology or plastic surgery institution. METHODS: In the preparatory phase (late 1995 and 1996), all general practitioners operating in the Friuli Venezia Giulia region (n = 1,038) were asked to participate in the program. Support from all regional dermatology, pathology and plastic surgery institutions was obtained. Operational procedures for the management of referred people were defined, and educational meetings directed to general practitioners interested in the program were held. Skin examinations by general practitioners started at the end of 1996 and took place during 1997 and 1998. Subsequently, information was obtained from participating general practitioners and from pathology institutions about the number and thickness of diagnosed melanomas, as well as the number of diagnosed skin carcinomas and dysplastic nevi. In addition, the thickness distribution of all melanomas diagnosed in the Friuli Venezia Giulia region before and during the program was obtained. RESULTS: A total of 153 general practitioners participated in the program, but only 74 were active and assessable. A total of 11,040 skin examinations was performed by these 74 general practitioners (median, 75 per general practitioner). In all, 820 people (7.4%) were referred for dermatological evaluation (median, 8 per general practitioner). Among these 820 people, at least 38 melanomas (4.6% of referred cases) were detected (18 < or = 1.5 mm, 11 > 1.5 mm thick, unknown in 9). The dermatological examinations/diagnosed melanomas ratio was 21. In addition, 94 skin carcinomas and 50 dysplastic nevi were detected. At the regional level, the percentage of thin melanomas rose from 65.3% in 1995-96 to 72.2% in 1997-98 (P = 0.04), whereas the number of thick melanomas declined. CONCLUSIONS: In our study, only a few general practitioners chose, in the absence of incentives, to participate in the study. However, the yield of melanomas, most of which were thin, was considerably high and the workload was acceptable. This compares favorably to experiences where dermatologists were involved directly without a filter work by general practitioners. PMID- 12729353 TI - Post-traumatic intracranial meningiomas. AB - The authors present 20 cases of likely post-traumatic intracranial meningiomas selected according to the conditions specified in the relevant literature. The relationship between head injury and subsequent development of meningioma remains a controversial and fascinating subject. The etiopathogenetic mechanisms and clinical features of our patients and those of the literature are discussed. PMID- 12729354 TI - Risk factors associated with genital warts in HIV-positive Brazilian women. AB - AIMS AND BACKGROUND: In Brazil, the female population has been increasingly infected with human immunodeficiency virus (HIV), and uterine cervix carcinoma now represents the second highest cause of mortality. Although HIV infection among women is associated with an increased prevalence of cervical cancer precursors, the co-infection with human papillomavirus (HPV) is considered to be a necessary but not sufficient factor to induce genital lesions. This study was conducted to identify risk factors associated with the history of genital warts among HIV-positive women. METHODS: A comparative cross-sectional evaluation was applied to 141 HIV-positive women. All patients were submitted to colposcopy, smear cytology, directed biopsy, and HPV-DNA detection. The chi-square, Fisher's exact test, and the odds ratio (OR, 95%; confidence interval, CI) were used to evaluate associations between history of genital warts and risk factors. RESULTS: A history of genital warts presented associations with: a) age at first sexual intercourse < or = 17 years (OR, 0.42; CI, 0.16-1.11); b) history of genital warts in sex partners (OR, 11.39; CI, 4.21-30.76), especially with recurrent episodes (OR, 6.60; CI, 2.69-16.12); c) drug addiction (OR, 2.38; CI, 1.09-5.19), especially in crack users (OR, 5.34; CI, 1.64-17.41); d) cervical HPV infection (OR, 2.75; CI 1.09-6.90); e) cervical infection caused by only one HPV type (OR, 2.77; CI 1.06-7.20); f) perianal HPV infection (OR, 2.30; CI, 0.70-7.56), associated with negative results for undetermined risk HPV (OR, 8.41; P = 0.04); and g) no antiretroviral therapy (OR, 3.41; P = 0.07). CONCLUSIONS: Evaluation of behavioral risk factors associated with a genital wart history is an important tool to prevent and reduce persistent HPV infection, and consequently genital cancer precursors in HIV infected women. PMID- 12729355 TI - Surgically proven complete response of stage III non-small cell lung cancer after cisplatin-enhanced radiotherapy. Clinical implications and long-term results. AB - AIMS AND BACKGROUND: Pathologic complete response in locally advanced non-small cell lung cancer is the main end point of combined therapies (chemotherapy and/or radiotherapy). Surgery after an induction treatment can improve local control, allowing the histologic assessment of treatment activity by means of resection or extensive biopsies. METHODS: Thirty patients surgically assessed without viable tumor after concurrent radiotherapy and continuous infusion of low-dose cisplatin, owing to an initially unresectable stage III non-small-cell lung cancer, were the object of evaluation to assess clinical implications, short- and long-term surgical results. RESULTS: The specificity rate of the preoperative restaging was 36.6%. The surgical procedures consisted of 22 resections and of extensive biopsies in 8 cases. The operative mortality was 4% (1/25) for procedures other than right pneumonectomy (3/5). No patient received postoperative chemotherapy. Eleven distant progressions, 4 local recurrences, and 4 new primary tumors were assessed as initial failures. The 8-year overall survival was 36%. CONCLUSIONS: Pathologic complete response after cisplatin enhanced radiotherapy cannot be satisfactorily assessed by clinical means. Surgery is required to obtain a reliable evaluation; however, right pneumonectomy should be contraindicated because of prohibitive risk. Although an effective local treatment can cure patients with advanced stage III disease, the addition of chemotherapy seems advisable to improve tumor relapse control. PMID- 12729356 TI - Compliance and outcomes in locally advanced head and neck cancer patients treated with alternating chemo-radiotherapy in clinical practice. AB - BACKGROUND AND AIMS: To evaluate the feasibility in clinical practice of alternating chemo-radiotherapy in locally advanced head and neck cancer patients. PATIENTS AND METHODS: From August 1993 to April 1998 at the Division of Medical Oncology of Parma, 48 consecutive patients were observed, and 38 (79%) started the Merlano chemo-radiotherapy. The characteristics of the patients were: males (32, 84%); median age, 57 years; PS <2 (32, 84%). The primary sites were the oropharynx (18, 47%), oral cavity (8, 21%), hypopharynx (7, 19%), larynx (5, 13%); stage IV disease was present in 29 (76%) patients. Twenty-five (66%) patients were married, and 24 (63%) resided outside of the city. RESULTS: The compliance was very low: 21 patients (55%) performed all the programmed cycles of chemotherapy, whereas only 5 patients (13%) performed the chemo-radiotherapy at full doses without any delay. The objective responses were 3 (8%) complete and 21 (55%) complete plus partial responses. Failures were 2 (5%) stable disease and 2 (5%) progressive disease, and the response was not assessable in 10 (26%). The median duration of the response was 8 months. The median overall survival and the time to progression were 18 and 13 months, respectively; the 5-year overall and relapse-free survival were 36% and 26%, respectively. Nine (24%) patients were still alive as of August 30, 2001, 8 (21%) of them without progression. Twenty six patients (68%) died with a local-regional relapse. One patient (3%) died for a second cancer. Grade 3-4 hematologic toxicity was leukopenia (n = 25, 66%) and thrombocytopenia (n = 9, 24%); grade 3-4 non-hematologic toxicity was diarrhea (n = 3, 8%) and mucositis (n = 2, 5%). Two patients (5%) died for intestinal infarction and perforation possibly related to treatment. CONCLUSIONS: Compliance to the chemo-radiotherapy was very poor. The response rate was lower than that reported in clinical trials, whereas overall survival was comparable. The alternating chemo-radiotherapy is a very complex treatment that cannot be easily applied in clinical practice; a careful selection of patients is mandatory not only considering oncologic and medical criteria, but also the level of awareness of the patient and his family. PMID- 12729358 TI - Peritoneal cystic mesothelioma: a case series. AB - BACKGROUND: Cystic peritoneal mesothelioma is a rare disease associated with a favorable short-term prognosis. Longer follow-up documenting a persistence of symptoms and a high rate of recurrence after debulking surgery along with an uncertain natural history prompt a re-evaluation of prior treatment recommendations. No prior long-term clinical study of these patients is available. METHODS: The experience with five cases of cystic peritoneal mesothelioma, four females and one male, are reviewed. All of these patients were treated with cytoreductive surgery with peritonectomy procedures and heated intraoperative intraperitoneal chemotherapy. CT, pathology and current status were investigated in order to learn more about the natural history of this disease. RESULTS: All patients were symptomatic from abdominal distention and three of the four complained of severe pain. Female patients complained of long periods of recurrent abdominal and pelvic pain poorly managed by oral analgesics. In one patient prolonged conservative management over ten years resulted in transition to an invasive process with extensive lymph nodal metastases. Her prognosis for long-term survival is guarded because of mesothelioma extension into the chest. Disease control of both ascites and pain in the abdomen and pelvis was achieved in all five patients treated with cytoreductive surgery plus intraperitoneal chemotherapy. CONCLUSIONS: Cystic peritoneal mesothelioma should no longer be referred to as "benign" cystic mesothelioma. An aggressive approach with complete disease eradication is the correct goal of treatment. From our experience, cytoreductive surgery to remove all visible tumor and intraperitoneal chemotherapy to control microscopic residual disease will help patients with peritoneal cystic mesothelioma to remain symptom- and disease-free over an extended time period with a single surgical intervention. Disease eradication may prevent the transition to an aggressive and fatal disease process. PMID- 12729357 TI - Vinorelbine and prednisone in older cancer patients with hormone-refractory metastatic prostate cancer. A phase II study. AB - AIMS AND BACKGROUND: Prostate cancer is a common disease in older men. Since it is hormone resistant, no treatment may improve survival. In patients with hormone refractory prostate cancer, clinical benefit is an important treatment end point. STUDY DESIGN: This study evaluated the efficacy and toxicity of a vinorelbine and prednisone combination in hormone-refractory prostate cancer patients. Vinorelbine was administered at the dose of 25 mg/m2 on days 1 and 8, every three weeks; prednisone was administered orally at the dose of 12 mg/day. Thirty consecutive patients, 65 years or older, with progressive (PSA increase or increase in bidimensionally measurable lesion) metastatic prostate adenocarcinoma were enrolled. Four patients (13%) had a partial response and 14 (46%) stable disease. Time to progression for the entire group was 4.5 months (range, 2-13) and 7.5 months for the group of responders (range, 3-13). A PSA decrease >50% was registered in 36% of the patients. Pain reduction was recorded in 44.4% of the patients and stability in 14.8%. RESULTS: The treatment was well tolerated and grade 3 toxicity was found in 2 cases of anemia and 2 cases of leukopenia without fever. CONCLUSIONS: The schedule is able to control the evolution of hormone refractory prostate cancer and to give a clinical benefit. These results provide information for further clinical trials in a large series of elderly cancer patients. PMID- 12729359 TI - Early and late outcome after surgery for colorectal cancer: elective versus emergency surgery. AB - AIMS AND BACKGROUND: Emergency surgery for colorectal cancer is associated with a higher postoperative morbidity and mortality rate and a poor long-term outcome compared with elective surgery. The aim of the present study was to compare early and late outcome after elective and emergency surgery for malignant colorectal cancer, looking for the principal determinants of a worse outcome after emergency colorectal surgery. METHODS: A retrospective study of 236 patients presenting with colorectal cancer over an 8-year period was undertaken. Of these, 118 presented as emergencies, whereas 118 patients, well matched for age, sex, site of tumor and TNM admitted as elective, were included in the study. Data reviewed included postoperative mortality and morbidity and long-term outcome. RESULTS: The 30-day operative mortality rate was significantly higher in the emergency group than in the electively treated group (11.9% versus 3.4%, P < 0.01). The higher mortality rate was observed in the perforation group. The 30-day operative morbidity was higher in the emergency group (27.1% versus 12.7%, P < 0.05). Anastomotic failure was a serious complication: following primary resection, we observed 4 non-fatal (5.4%) and two fatal (2.7%) anastomotic leaks after 74 primary anastomoses. Among emergency-treated patients, the procedures characterized by the highest percentage of postoperative complications were three stage resections (63.6%). The 5-year survival rate was greater after elective surgery (59% versus 39%). CONCLUSIONS: The early and long-term outcome following emergency colorectal surgery was significantly lower than that after elective surgery. Although medical complications in patients with end-stage cancer played an important role, surgical failures still had an important impact on outcome. PMID- 12729360 TI - Immediate effects of intraoperative evaluation of surgical margins over the treatment of early infiltrating breast carcinoma. AB - AIMS AND BACKGROUND: Local recurrences in patients submitted to conservative breast treatment for early infiltrating breast carcinomas occur in 5-10% of the cases and are caused mainly by inadequate local resection and remaining residual malignant tissue. The present study was carried out to analyze the effect of intraoperative evaluation of surgical margins and its influence on the immediate surgical management of patients with early breast carcinomas (T1-T2) scheduled to undergo quadrantectomy. METHODS: A total of 102 cases were studied. After a classical quadrantectomy, intraoperative evaluation of surgical margins was done by means of macroscopic, cytological and histopathologic analysis. The margins of the resected tissue were examined to assure they were clear or to orient a wider resection. RESULTS: In 64 cases (62.7%), the extent of the quadrant resection was considered adequate and the margins were clear. In 38 cases (37.3%), surgical margins were considered inadequate. An enlarged quadrantectomy was immediately performed in 33 patients (32.4%) and mastectomies in 5 (4.9%). CONCLUSIONS: Intraoperative evaluation of surgical margins frequently modifies the surgical management of patients who were initially prepared to be submitted to a quadrantectomy, indicating the need for further resection in the form of an enlarged quadrantectomy or mastectomy. PMID- 12729361 TI - HMB45 and melan-A expression in renal angiomyolipoma and their significance for the diagnosis. AB - AIMS AND BACKGROUND: The melanosome-associated proteins, also called HMB45 and melan-A, are also present in renal angiomyolipoma. The aim of the present study was to evaluate the expression of HMB45 and melan-A in mesenchymal cells of renal angiomyolipoma and to investigate their significance in the differential diagnosis. METHODS: Twelve patients, 9 females and 3 males diagnosed with renal angiomyolipomas, were included in the present study. The most representative tumor tissue block was chosen from each case, and 5-microm sections were taken to poly-L-lysin-coated slides for immunohistochemical staining. The standard streptavidin-biotin immunoperoxidase method was used for immunostaining with HMB45 and melan-A antibodies. RESULTS: All of the cases showed positive cytoplasmic immunostaining for HMB45 and melan-A. Melan-A expression was shown in smooth muscle component, adipose tissue and predominantly in the perivascular cells, whereas HMB45 immunoreactivity was stronger than melan-A expression in all cases. CONCLUSIONS: It was concluded that HMB45 and melan-A reactivity is a useful tool to distinguish renal angiomyolipomas from other primary and secondary mesenchymal and primary epithelial tumors. Melan-A and HMB45 share similar specificities for renal angiomyolipoma. In addition, such expression in renal angiomyolipomas may occur without any evidence of nevomelanocytic differentiation. Further research is required to determine the histogenesis of this entity. PMID- 12729362 TI - Neuroendocrine differentiation in colorectal carcinomas: assessing its prognostic significance. AB - AIMS AND BACKGROUND: There is evidence that colorectal carcinomas with extensive neuroendocrine features have a substantially worse prognosis than those without, but the frequency and clinical significance of neuroendocrine features in conventional carcinomas has not been settled since few studies have been performed, with conflicting results. The aim of the study was to investigate neuroendocrine differentiation in colorectal carcinomas in relation to its prognostic significance. METHODS: In 50 patients with colorectal carcinoma, the extent and intensity of staining with each of the three antibodies (chromogranin A, neuron-specific enolase and synaptophysin) were determined and correlated with the histologic type, grade, stage of the tumor and survival time ot the patients. RESULTS: We observed chromogranin A expression in 38%, neuron-specific enolase expression in 26%, and synaptophysin expression in 6% of the tumors. Chromogranin A was the most frequently and strongly expressed marker in our study. Of the three antibodies studied, only chromogranin A positivity was correlated with grade and stage of the tumors and was associated with a decreased effect on survival. CONCLUSIONS: Our results show that chromogranin A is the most sensitive and specific neuroendocrine marker. Chromogranin A positivity appears to bear a poor prognosis in patients with colorectal cancers. PMID- 12729363 TI - Expression of Bcl-2, Bax and p53 proteins in pituitary adenomas: an immunohistochemical study. AB - AIMS AND BACKGROUND: Although pituitary adenomas are usually benign lesions, their growth rate is highly variable and unpredictable. Apoptosis appears to be an important process in neoplastic lesions. The purpose of this study was to investigate the expression of apoptosis-related proteins including Bcl-2, bax and p53 in pituitary adenomas and its correlation with hormone function, tumor size, local control, and proliferative activity. STUDY DESIGN: The expression of Bcl-2, Bax and p53 proteins and hormonal function were determined in formalin-fixed, paraffin-embedded tissue from 41 untreated pituitary adenomas using immunohistochemistry. The patients were followed for a median of 60 months (range, 12 to 95). Patient charts were reviewed to record tumor recurrence and size. Tumor proliferative activity was assessed by immunohistochemistry using Ki 67 antibody. RESULTS: Of 41 pituitary adenomas, 26 (63%) were hormone-secreting and 15 (37%) non-functioning, 34 (83%) were macroadenoma and 7 (17%) microadenoma, and 15 (37%) showed local relapse. Six (14%) adenomas were of low proliferative activity, whereas the others (86%) were non-proliferative. Immunohistochemically, 31 adenomas (75%) showed bcl-2 positivity, 37 (90%) bax positivity, and 7 (17%) p53 positivity. Statistical analysis revealed that Bcl-2 protein expression significantly diminished in prolactin-secreting and non functioning adenomas (P = 0.005 and P = 0.006, respectively), and increased in growth hormone-secreting adenomas (P = 0.003). In addition, expression of bax protein significantly decreased in recurrent tumors, in contrast to p53 protein, which showed a significant increase (P = 0.03 and P = 0.002, respectively). CONCLUSIONS: We think that apoptosis-related proteins such as Bcl-2, Bax and p53 may be significantly related to hormone function and local control in pituitary adenomas. PMID- 12729364 TI - Analysis of irradiated lung and heart volumes using virtual simulation in postoperative treatment of stage I breast carcinoma. AB - AIMS AND BACKGROUND: The aim of the study was to assess the usefulness of virtual simulation in postoperative radiotherapy treatment planning of early-stage breast cancer and to evaluate its potential to reduce the volume of critical structures exposed compared to treatment plans produced by a conventional 2D system. METHODS AND STUDY DESIGN: Eighteen patients undergoing breast radiotherapy following conservative surgery for small breast carcinomas were studied. Scans from spiral CT equipment (with the patient in the treatment position) were transferred to a virtual simulator. From the screen images the operator contoured breast, lung and heart. Calculations were made of the extent to which the heart and lung were included in the irradiation fields (50% isodose line of tangential fields). RESULTS: Manual contouring was time-consuming, but when virtual simulation was used, the mean volume of the lung included in the radiation fields was significantly reduced compared to the 2D treatment plan (4.5% vs 5.4%, P = 0.034); in addition, a slight reduction was observed for the heart (0.5% to 1.2%), but this was not statistically significant. CONCLUSIONS: With a 3D system we obtained optimal target coverage and a reduction of the dose to critical structures (statistically significant only for the lung). From a clinical point of view, this 0.9% reduction in the mean irradiated lung volume is probably not significant, as the percentage irradiated with a 2D system is considerably below the recommended value. Furthermore, our analysis was performed in a relatively small group of patients; for a reliable estimate larger series would be required. Consequently, the 3D system should not be considered in routine treatment after breast conserving surgery for early stage carcinomas; for the time being it should be reserved for selected cases. PMID- 12729365 TI - Late visual and auditory toxicity of radiotherapy for nasopharyngeal carcinoma. AB - AIMS AND BACKGROUND: Classical irradiation of carcinoma of the nasopharynx involves large fields and high doses; therefore, significant late toxicity and late side effects are to be expected. Given the fact that nasopharyngeal carcinoma (NPC) is a relatively radiosensitive disease and a significant proportion of patients are long-term survivors, late visual and auditory complications of treatment are of utmost concern for patients and radiation oncologists. The aim of this study was to evaluate the long-term visual and auditory toxicity in patients treated with radiotherapy for NPC. METHODS AND STUDY DESIGN: Forty-three long-term survivors (including 11 children), following definitive radiotherapy for NPC, underwent a thorough visual and auditory evaluation 2-22 years after their treatment. Ophthalmological examination consisted of anamnesis of dry eye syndrome and visual acuity, visual acuity testing, slit-lamp examination, Schirmer test, fundus examination, and intraocular pressure measurement, as well as fluorescein angiography in patients with pathological vascular findings in the fundus examination. Audiological evaluation included anamnesis of hearing loss, tinnitus or vertigo, examination of the ears and nasopharynx, audiogram, and tympanogram. RESULTS: Radiation retinopathy was found in 16% of patients by fundus examination, with one patient (2.3%) developing blindness. Severe dry eye syndrome was present in 26%. Fifty six percent had some degree of hearing impairment, with 74% showing severe sensorineural hearing loss. Fifty-eight percent of patients reported tinnitus and 26% reported suffering from dizziness. Radiation retinopathy as well as all manifestations of auditory toxicity were found to bear a direct correlation with dose per fraction. CONCLUSIONS: Unless there is tumor involvement, the orbital contents should be completely excluded from the target volume. Auditory toxicity is significant when treating NPC with two-dimensional techniques. PMID- 12729366 TI - Radiotherapy for medically inoperable non-small cell lung cancer at clinical stage I and II. AB - We reviewed the records of 36 patients with medically inoperable stage I-II non small cell lung carcinoma who were treated with radiotherapy. The median dose to the target was 60 Gy with conventional fractionation. Fifteen patients were treated without elective irradiation fields, while the remaining 21 were treated with extended fields including elective mediastinal regional lymph nodes. The overall survival rates at three and five years were 32.3% and 18.8%, the cause specific survival rates were 40.9% and 27.3%, and the local control rates were 31.7% and 23.8%, respectively. In multivariate analysis the radiation dose had a marginally significant influence on the cause-specific survival, while tumor size had a significant influence on the local control rate. Only one patient had relapse in the regional mediastinal lymph nodes as the only site of metastasis. We conclude that the dose used in the present study is inadequate and recommend that further efforts be made to improve local control by dose escalation within a small target volume. PMID- 12729367 TI - Cisplatin treatment triggers familial Mediterranean fever attacks. AB - A 42-year-old familial Mediterranean fever (FMF) patient who was treated with cisplatin-based chemotherapy for adenocarcinoma of the lung developed severe and frequent attacks of FMF during treatment. Abdominal pain, arthralgia and fever occurred for a few days following each cisplatin cycle. His FMF worsened, the abdominal pain and fever lasted longer and treatment with colchicine was ineffective. It has been hypothesized that the link between cisplatin treatment and FMF attacks lies in an increased production of serotonin, IL-6, IL-1, IL-8 and TNF-alpha. These inflammatory cytokines have been reported to be overproduced during cisplatin treatment and are known to play an important role in FMF relapse. The oncologist should be made aware of the possibility of disease aggravation in FMF patients during cisplatin-based chemotherapy. PMID- 12729368 TI - Complete response after chemoradiation in ampullary carcinoma: a case report. AB - AIMS AND BACKGROUND: The case of a 70-year-old patient with resectable, poorly differentiated adenocarcinoma of the ampulla of Vater is presented. PATIENT AND METHODS: Due to intraoperative hemorrhagic complications, surgical resection was not feasible. The patient was treated with radiochemotherapy consisting of external beam radiotherapy (50.4 Gy; 1.8 Gy/fraction; 5 fractions/week) plus 5-FU (1000 mg/m2/day, continuous i.v. infusion, days 2-5, week 1 and 5 of radiotherapy) and mitomycin C (10 mg/m2 i.v., day 2, week 1 of radiotherapy). RESULTS: At five years' follow-up the patient was in good general condition, without any signs of disease according to CT scan, endoscopic retrograde cholangiopancreatography and tumor marker determination. Multiple random biopsies performed in the ampullary region were negative for tumor growth. CONCLUSIONS: In patients with ampullary carcinoma the use of concurrent chemoradiation should be considered, particularly when surgical resection is unfeasible due to medical contraindications or locally advanced disease. PMID- 12729369 TI - Responsiveness of skin metastases to CMF in a patient with urothelial carcinoma of the bladder: a case report. AB - Skin metastases from urothelial carcinoma of the bladder are uncommon, and there are few cases reported in literature. The present case report describes the results of a combination of cyclophosphamide, methotrexate and 5-fluorouracil (CMF) administered as second-line chemotherapy in a cisplatin-resistant metastatic bladder cancer patient. The improvement in cutaneous lesions and pain reduction obtained prompt further exploration of the activity of this regimen in a second-line approach. PMID- 12729370 TI - Primary rectal lymphoma as a cause of bowel obstruction. Report of a case. AB - BACKGROUND: Malignant lymphoma involving the rectum either as a localized process or as a manifestation of disseminated disease is rare. Several treatments have been proposed and reported, including surgical resection alone or associated with adjuvant chemoradiation, chemotherapy alone, and radiotherapy alone. METHODS: A case of bowel obstruction caused by a primary rectal MALT lymphoma is reported. Following emergency loop sigmoid colostomy the patient was started on multiple specific cycles of chemotherapy according to the MACOP-B protocol. RESULTS: At the end of chemotherapy a remarkable reduction in the size of the tumor was noted. Subsequently the patient underwent an ultralow anterior resection followed by a straight coloanal anastomosis. At 36 months of follow-up the patient is alive with no tumor recurrence. CONCLUSIONS: The present report describes the unique case of a patient with primary obstructing rectal lymphoma treated with neoadjuvant chemotherapy and sphincter-saving curative surgery. PMID- 12729371 TI - Angioimmunoblastic lymphadenopathy with dysproteinemia following doxycycline administration. AB - Angioimmunoblastic lymphadenopathy with dysproteinemia (AILD) is a primary lymphoproliferative T-cell disorder, currently classified as a peripheral T-cell non-Hodgkin's lymphoma. AILD is characterized by generalized lymphadenopathy, hepatosplenomegaly, immunological abnormalities, polyclonal hypergammaglobulinemia and anemia. We report a case of AILD in an 80-year-old male who presented with a generalized pruritic maculopapular eruption and fever following doxycycline administration. The maculopapular rash progressed to formation of confluent nodules, plaques and finally erythroderma with lymphadenopathy and hepatosplenomegaly. Blood analysis revealed an elevated erythrocyte sedimentation rate and polyclonal hypergammaglobulinemia. Lymph node biopsy showed almost complete effacement of the nodal architecture with diffuse proliferation of small vessels forming an arborizing network, surrounded by atypical lymphocytes, usually CD3+ CD4+ and occasionally CD3+ CD8+. There were also larger cells (immunoblastic shape) that displayed CD20 positively, some scattered plasma cells, and eosinophils. Histology of a cutaneous lesion showed spongiosis and infiltration of the epidermis by atypical lymphocytes with large hyperchromatic nuclei, perivascular dermal lymphocytic infiltrate (CD3+) mixed with plasma cells and occasional large immunoblasts (CD20+). During hospitalization the patient developed hemolytic anemia (Coombs positive) and lung metastases. The prognosis of AILD is generally poor, with a median survival of less than 20 months. Our patient died two and a half months after the diagnosis was made due to sepsis caused by Staphylococcus aureus isolated in hemoculture. PMID- 12729372 TI - Solitary intracranial subdural osteoma: case report and review of the literature. AB - Intracranial subaural osteomas without any relation to osseous or meningeal tissues are rare, with only five cases including the present one having been reported so far. In most patients headache, which was always localized at the site of the lesion, was the most frequent complaint and subsided after complete resection of the neoplasm. The mechanism of origin of such lesions remains unknown. PMID- 12729373 TI - Renal metastasis from thyroid carcinoma 35 years after detection of the primary tumor. AB - A case of a solitary, unilateral, large metastatic renal tumor found 35 years after the detection of primary thyroid carcinoma is presented. Renal metastasis from thyroid carcinoma detected during life is extremely rare, and there may be a long latency period before the appearance of a secondary tumor. The possibility of a secondary tumor should always be considered in the differential diagnosis of renal neoplasms, even when the tumor is a large, unilateral, solitary renal mass. PMID- 12729374 TI - Primary pulmonary meningioma: report of a case and review of the literature. AB - Primary pulmonary meningiomas are rare and their occurrence has been reported only sporadically. The diagnosis of such tumors should be accepted only after the presence of an intracranial or intraspinal meningioma has been excluded. The morphological and immunohistochemical features are similar to those of intracranial meningiomas. A case of primary pulmonary meningioma along with a review of the literature and discussion of other lesions in the differential diagnosis of this rare neoplasm are here presented. PMID- 12729375 TI - Metastatic carcinoma of the breast mimicking multiple myeloma. PMID- 12729376 TI - [New insights into the combinability of Eloxation (oxaliplatin): present and future prospects]. PMID- 12729377 TI - Delusions. PMID- 12729378 TI - Luminance gradient can break background-independent lightness constancy. AB - A display with a luminance gradient was shown to induce a strong lightness illusion (Logvinenko, 1999 Perception 28 803-816). However, a 3-D cardboard model of this display was found to produce a much weaker illusion (less than half that in the pictorial version) despite the fact that its retinal image is practically the same. This is in line with the hypothesis that simultaneous lightness contrast is solely a phenomenon of pictorial perception (Logvinenko et al, 2002 Perception 31 73-82). The residual lightness illusion in the 3-D model can be accounted for by the fact that this model is a hybrid display. Specifically, while it is a real object, a pictorial representation (of the illumination gradient) is superimposed on it. Thus, lightness in the 3-D display is a compromise between two opposite tendencies: the background-independent lightness constancy and the lightness illusory shift induced by the luminance gradient. PMID- 12729379 TI - Perceptual organization and White's illusion. AB - The apparent lightness of a surface can be strongly modulated by the spatial context in which it is embedded. Early theories of such context dependence emphasized the role of low-level mechanisms that sense border contrast, whereas a number of recent authors have emphasized the role of perceptual organization in determining perceived lightness. One of the simplest and most theoretically challenging lightness illusions was described by White. This illusion has been explained with a variety of different models, ranging from low-level filter outputs to computations underlying the extraction of mid-level representations of surfaces. Here, I present a new method for determining the organizational forces that shape this illusion. I show that the spatial context of White's pattern not only transforms the apparent lightness of homogeneous target patches. but can also induce dramatic inversions of figure-ground relationships of textured target regions. These phenomena provide new evidence for the role of scission in causing the lightness illusion experienced in White's effect. PMID- 12729380 TI - The role of eyebrows in face recognition. AB - A fundamental challenge in face recognition lies in determining which facial characteristics are important in the identification of faces. Several studies have indicated the significance of certain facial features in this regard, particularly internal ones such as the eyes and mouth. Surprisingly, however, one rather prominent facial feature has received little attention in this domain: the eyebrows. Past work has examined the role of eyebrows in emotional expression and nonverbal communication, as well as in facial aesthetics and sexual dimorphism. However, it has not been made clear whether the eyebrows play an important role in the identification of faces. Here, we report experimental results which suggest that for face recognition the eyebrows may be at least as influential as the eyes. Specifically, we find that the absence of eyebrows in familiar faces leads to a very large and significant disruption in recognition performance. In fact, a significantly greater decrement in face recognition is observed in the absence of eyebrows than in the absence of eyes. These results may have important implications for our understanding of the mechanisms of face recognition in humans as well as for the development of artificial face-recognition systems. PMID- 12729382 TI - Cognitive styles: errors in directional judgments. AB - Previous studies on spatial memory have shown that, in judging direction, participants are more accurate and faster when a map is aligned with the perspective of the spatial layout they had learned (alignment effect). Rossano and Warren (1989 Perception 18 215-229) have shown that when participants have to do a contra-aligned judgment they can either answer correctly, or make alignment or mirror-image errors. We think that the kind of response depends on the different way in which people acquire environmental knowledge: landmark, route, and survey. We hypothesise that landmark and route participants show alignment effects and make, respectively, alignment errors and mirror-image errors, whereas survey participants do not show an alignment effect. An experiment is reported in which participants performed three tasks in order to distinguish their cognitive style. We selected thirty landmark, thirty route, and twenty-eight survey participants. They were then submitted to directional judgment tasks to verify whether the alignment effect was present and to observe the kind of responses. The results revealed that survey participants did not show an alignment effect, and that the kind of errors could depend on the directional judgment task participants had to do, and not only on the cognitive style. PMID- 12729381 TI - A common oscillator for perceptual rivalries? AB - Perceptual rivalry is an oscillation of conscious experience that takes place despite univarying. if ambiguous, sensory input. Much current interest is focused on the controversy over the neural site of binocular rivalry, a variety of perceptual rivalry for which a number of different cortical regions have been implicated. Debate continues over the relative role of higher levels of processing compared with primary visual cortex and the suggestion that different forms of rivalry involve different cortical areas. Here we show that the temporal pattern of disappearance and reappearance in motion-induced blindness (MIB) (Bonneh et al, 2001 Nature 411 798-801) is highly correlated with the pattern of oscillation reported during binocular rivalry in the same individual. This correlation holds over a wide range of inter-individual variation. Temporal similarity in the two phenomena was strikingly confirmed by the effects of the hallucinogen LSD, which produced the same, extraordinary, pattern of increased rhythmicity in both kinds of perceptual oscillation. Furthermore. MIB demonstrates the two properties previously considered characteristic of binocular rivalry. Namely the distribution of dominance periods can be approximated by a gamma distribution and, in line with Levelt's second proposition of binocular rivalry, predominance of one perceptual phase can be increased through a reduction in the predominance time of the opposing phase. We conclude that (i) MIB is a form of perceptual rivalry, and (ii) there may be a common oscillator responsible for timing aspects of all forms of perceptual rivalry. PMID- 12729383 TI - Drawings from Gaia, a blind girl. AB - Gaia, a totally blind girl, was asked to make raised-line drawings. Gaia's vision at best was peripheral. She draws out of interest, and has drawn since preschool with encouragement from her mother. She was asked to draw objects and scenes involving depth from a vantage point, eg a table from below, two cars (one behind the other), and two parallel rows of apples (receding from her, on a table top). Gaia represented space in her drawings using T-junctions for overlap, height in the picture plane, parallel projection, and inverse projection. That is, Gaia uses features of systems common in sighted children's drawings. The development of drawing in blind and sighted children may be similar in good measure because haptics provides access to many of the same spatial principles as vision. PMID- 12729384 TI - The formation of rhythmic categories and metric priming. AB - Two experiments on categorical rhythm perception are reported, the object of which was to investigate how listeners perceive discrete rhythmic categories while listening to rhythms performed on a continuous time scale. This is studied by considering the space of all temporal patterns (all possible rhythms made up of three intervals) and how they, in perception, are partitioned into categories, ie where the boundaries of these categories are located. This process of categorisation is formalised as the mapping from the continuous space of a series of time intervals to a discrete, symbolic domain of integer-ratio sequences. The methodological framework uses concepts from mathematics and psychology (eg convexity and entropy) that allow precise characterisations of the empirical results. In the first experiment, twenty-nine participants performed an identification task with 66 rhythmic stimuli (a systematic sampling of the performance space). The results show that listeners do not just perceive the time intervals between onsets of sounds as placed in a homogeneous continuum. Instead, they can reliably identify rhythmic categories, as a chronotopic time clumping map reveals. In a second experiment, the effect of metric priming was studied by presenting the same stimuli but preceded with a duple or triple metre subdivision. It is shown that presenting patterns in the context of a metre has a large effect on rhythmic categorisation: the presence of a specific musical metre primes the perception of specific rhythmic patterns. PMID- 12729385 TI - The aperture problems in the Pulfrich effect. AB - The Pulfrich effect yields a perceived depth for horizontally moving objects but not for vertically moving ones. In this study the Pulfrich effect was measured by translating oblique lines seen through a circular window, which made motion direction ambiguous. Overlaying random dots that moved horizontally, vertically, or diagonally controlled the perceptual motion direction of the lines. In experiment 1, when the lines were seen to move horizontally, the effect was strongest in spite of the same physical motion of the lines. Experiment 2 was performed to test the above conditions again, excluding the Pulfrich effect of the dots on the depth of the lines. The overlaid dots were presented to one eye only. The result showed that the Pulfrich effect of the lines was persistently strong in spite of the perceptual changes in motion direction. Experiment 3 also showed that the Pulfrich depth was independent of the perceived horizontal speed in a plaid display. The Pulfrich effect was determined by measuring the horizontal disparity component, independently of the perceived motion direction. These results demonstrate that the aperture problems in motion and stereopsis in the Pulfrich effect are solved independently. PMID- 12729386 TI - A chimeric point-light walker. AB - Ambiguity has long been used as a probe into visual processing. Here, we describe a new dynamic ambiguous figure-the chimeric point-light walker--which we hope will prove to be a useful tool for exploring biological motion. We begin by describing the construction of the stimulus and discussing the compelling finding that, when presented in a mask, observers consistently fail to notice anything odd about the walker, reporting instead that they are watching an unambiguous figure moving either to the left or right. Some observers report that the initial percept fluctuates, moving first to the left, then to the right, or vice versa; others always perceive a constant direction. All observers, when briefly shown the unmasked ambiguous figure, have no difficulty in perceiving the novel motion pattern once the mask is returned. These two findings--the initial report of unambiguous motion and the subsequent 'primed' perception of the ambiguity--are both consistent with an important role for top-down processing in biological motion. We conclude by suggesting several domains within the realm of biological motion processing where this simple stimulus may prove to be useful. PMID- 12729387 TI - Treatment of microarray experiments as split-plot designs. AB - This paper shows that microarray experiments are split-plot, or split-unit, designs. The larger size experimental unit (the whole plot) is the array, and the treatment applied to this unit is the treatment given to the cells which produce the cDNA that is hybridized to the array. The smaller size experimental unit (the subplot) is the spot on the array, and the treatment applied to this unit is the gene giving rise to the DNA or oligonucleotide attached at that spot. Various treatment and design structures can be applied to the whole plot and the subplot; we consider the model equations appropriate to different designs. Preliminary normalization of the data can be avoided by including appropriate blocking terms in the model equation. We show how conventional analysis of variance can be used to test for significant differences in expression, and consider multiplicity corrections and graphical methods for identifying important expression differences. PMID- 12729388 TI - Assessing response profiles from incomplete longitudinal clinical trial data under regulatory considerations. AB - Treatment effects are often evaluated by comparing change over time in outcome measures. However, valid analyses of longitudinal data can be problematic, particularly when some data are missing for reasons related to the outcome. In choosing the primary analysis for confirmatory clinical trials, regulatory agencies have for decades favored the last observation carried forward (LOCF) approach for imputing missing values. Many advances in statistical methodology, and also in our ability to implement those methods, have been made in recent years. The characteristics of data from acute phase clinical trials can be exploited to develop an appropriate analysis for assessing response profiles in a regulatory setting. These data characteristics and regulatory considerations will be reviewed. Approaches for handling missing data are compared along with options for modeling time effects and correlations between repeated measurements. Theory and empirical evidence are utilized to support the proposal that likelihood-based mixed-effects model repeated measures (MMRM) approaches, based on the missing at random assumption, provide superior control of Type I and Type II errors when compared with the traditional LOCF approach, which is based on the more restrictive missing completely at random assumption. It is further reasoned that in acute phase clinical trials, unstructured modeling of time trends and within subject error correlations may be preferred. PMID- 12729389 TI - Covariate-adjusted reference intervals for diagnostic data. AB - The analysis of extreme diagnostic measurements in clinical trials relies on reference intervals that help drug developers quickly determine whether a particular value is typical or atypical. The distribution of diagnostic variables is often greatly influenced by various covariates and it is important to properly account for this influence in the analysis of extreme measurements. This paper discusses three approaches to constructing covariate-adjusted reference intervals for quantitative diagnostic data: global quantile smoothing, local quantile smoothing, and stepwise quantile approximations based on recursive partitioning. A detailed review of methods for optimizing the quantile estimation procedures is provided. The paper presents algorithms for selecting the degree of a polynomial approximation in global smoothing, bandwidth parameter in local smoothing, and number of strata in recursive partitioning. The described methods for computing covariate-adjusted reference intervals are applied to the analysis of electrocardiographic data. PMID- 12729391 TI - Assessing the impact of endpoint shopping on power in confirmatory clinical trials. AB - When planning a confirmatory study, one of the important aspects is choosing a primary endpoint and evaluating power to show a difference. Data from preliminary studies are generally used for such planning. It is natural to want to use the endpoint from the preliminary study that best differentiates between test drug and control as the primary endpoint in the confirmatory study. However this leads to the possibility of bias in estimation of the effect size in the preliminary study, and, hence, lower than anticipated power in the confirmatory study. In this paper we quantify the impact of such endpoint shopping on the power of confirmatory studies. We find the upper bound on bias and show that for low to moderate correlation it is not very conservative. We derive the asymptotic distribution of the treatment effect and propose a test forequal treatment effects for the data from the preliminary study when endpoints are correlated. We study properties of this test. We propose a strategy to use the data from the preliminary study to plan confirmatory studies with unbiased or conservative estimates of power. PMID- 12729390 TI - Further developments of the Fisher information matrix in nonlinear mixed effects models with evaluation in population pharmacokinetics. AB - We extend the development of the expression of the Fisher information matrix in nonlinear mixed effects models for designs evaluation. We consider the dependence of the marginal variance of the observations with the mean parameters and assume an heteroscedastic variance error model. Complex models with interoccasions variability and parameters quantifying the influence of covariates are introduced. Two methods using a Taylor expansion of the model around the expectation of the random effects or a simulated value, using then Monte Carlo integration, are proposed and compared. Relevance of the resulting standard errors is investigated in a simulation study with NONMEM. PMID- 12729392 TI - Chi-square test for R x C contingency tables with clustered data. AB - In ophthalmologic or dental studies, observations are frequently taken from multiple sites (called units), such as eyes or teeth, of each subject. In this case, observations within each subject (called clusters) may be dependent, although those from different subjects are independent. When a categorical observation is made from each site, application of the usual Pearson chi-square tests is invalid since sites within the same subject tend to be dependent. We propose a modified chi2 statistic for testing no treatment effect in these cases. The proposed methods do not require correct specification of the dependence structure within cluster. Simulation studies are conducted to show the finite sample performance of the new methods. The proposed methods are applied to real life data. PMID- 12729393 TI - Analysis of survival data with missing measurements of a time-dependent binary covariate. AB - The objective of this study was to investigate the influence of the number and timing of a binary time-dependent covariate on the bias using the last observation carried forward in the proportional hazards model. Under various assumptions of censoring rates, transition probabilities of the time-dependent covariate, sample size, and the log hazard-ratio for the covariate, we empirically examined the impact that the number and timing have on the bias of the estimator of the covariate. An example from the Systolic Hypertension in the Elderly Program was used. Inference on the effect of systolic blood pressure on survival is strongly affected by the number and timing of systolic blood measurements. PMID- 12729394 TI - Bivariate longitudinal model for the analysis of the evolution of HIV RNA and CD4 cell count in HIV infection taking into account left censoring of HIV RNA measures. AB - We present a bivariate linear mixed model taking into account censored measures of the response variable due to lower quantification limit of the assays. It allows an estimate of the correlation between the two response variables and takes into account this correlation for the estimation of other model parameters. This model was applied in a large cohort study (APROCO Cohort) to study the evolution under antiretroviral treatment of the two major biomarkers of the progression of Human Immunodeficiency Virus (HIV) infection: plasma HIV RNA and CD4+ T lymphocytes cell count. In a sample of 929 patients who started an highly active antiretroviral therapy, we illustrate the superiority in terms of likelihood of a bivariate model compared to two univariate models and the impact of taking into account the left-censoring of HIV-RNA. Moreover, interpretation of the model parameters allows confirmation of correlation between these two markers throughout the whole follow-up and the continuous decrease of plasma HIV RNA on average. Despite some limitations (distribution assumption, ignorance of missingness process), such a model appeared to be very useful to correctly describe the current evolution of important biomarkers in HIV infection. PMID- 12729395 TI - Using a serial marker to predict a repeated measures outcome in a cohort study. AB - Consider the cohort design and suppose that the outcome of primary interest is a continuous random variable observed repeatedly over time. Suppose that the value of a "clinical marker," which is thought to be predictive of the primary outcome, is also recorded. We would like to determine whether there is an association between the two variables as they evolve over time. We might also want to predict the pattern for the primary outcome conditionally on a specific profile of clinical interest for the serial marker. A model is developed to address these issues. One regression model is created for the primary outcome while a second regression model is developed for the clinical marker. The vector autoregressive model, i.e., VAR(1), is used to characterize the covariance structure between the two sets of repeated observations. Hypotheses of interest are described and procedures for testing them are elaborated. The Diabetes Control and Complications Trial is used to illustrate the procedures. PMID- 12729397 TI - A longitudinal approach for constructing beta-expectation tolerance intervals. AB - A new method is presented for constructing beta-expectation tolerance limits (TLs) for longitudinal data with error components regression structure. The TLs are mainly intended for small samples where the variation between and within subjects is large. In such cases the new method is superior to existing methods, which are based on a cross-sectional approach and which do not utilize the longitudinal structure of the data. Simulation studies show that the mean length of the TLs can be much reduced by using the new approach, while at the same time the beta-expectation property is maintained. The gain from using the longitudinal approach furthermore increases with the beta-expectation level. The results are demonstrated on data consisting of measurements of glutamate concentrations in brains from rats. Here, the cross-sectional approach due to Wilks give 90-99% TLs, which are up to 101-230% wider than are those obtained with the new approach. PMID- 12729396 TI - Analyzing randomized dose finding studies with a primary and a secondary endpoint. AB - The identification of the minimum effective dose for both the primary and secondary endpoint under a priori importance ordering assumption is described. Stepwise testing procedures, which conditionally test each elementary hypotheses at level alpha, but control the experimentwise Type I error rate, can be used. PMID- 12729398 TI - Effect of within-household reinfestation on design sensitivity. AB - The effect of within-household reinfestation on design sensitivity is investigated through statistical modeling of the reinfestation process. When reinfestation is present, household randomization tends to magnify treatment differences when compared with individual randomization or a randomized blocking design in the simple setting of two patients per household. The effect of reinfestation under more general household randomization settings is investigated by determining the relationship between the treatment effect and the number of patients per household. In an actual clinical study of treatment for head lice infestation, household randomization with proper stratification was adopted. The results from the study were consistent with theoretical expectations. This seeming contradiction to the blocking principle demonstrates the need to check whether all conditions are met before applying standard design principles. PMID- 12729400 TI - Thyroid function and puberty. AB - Thyroid hormones are essential for normal growth, sexual development and reproductive function. During puberty, changes in thyroid functions and an increase in thyroid volume occur as an adaptation to body and sexual development. Hypothyroidism diagnosed late in prepubertal years, usually due to Hashimoto's thyroiditis, can cause a delay of puberty or incomplete isosexual precocity (development of breast and internal genitalia in girls and increased testis volume in boys without adrenarche). In contrast, normal pubertal development and adequate menarche have been documented in congenital hypothyroidism detected by neonatal screening and treated early. The effect of hyperthyroidism on pubertal development is not well known, but a short period of hyperthyroidism seems not to have major negative effects. In adolescence or young adulthood, menstrual dysfunction, infertility, and stillbirth or premature birth are associated with thyroid dysfunction. PMID- 12729399 TI - Chronic conditions and adolescence. AB - Globally, one in ten adolescents suffers from a chronic condition. This paper reviews the issues of adolescent development, family relationships, school performance, and risk behaviors among chronically ill adolescents using a non categorical approach. PMID- 12729401 TI - Growth and puberty in thalassemia major. AB - Present transfusional regimen protocols increase the life expectancy of patients with beta-thalassemia major, but cause a progressive iron overload that can be prevented or limited only by appropriate iron chelation. Siderosis is responsible for the clinical complications of the disease. Short stature and hypogonadism are extremely frequent in patients with thalassemia. Many factors are responsible for short stature in patients with thalassemia, the most important of which are dysfunction of the GH-IGF-I axis and desferoxamine (DFX)-induced bone dysplasia. Hypogonadism is the most frequent endocrine complication, mostly due to gonadotrophins deficiency secondary to iron overload. Sex steroid treatment for induction of puberty and/or maintenance of sexual characteristics is necessary. Both short stature and hypogonadism are present in a significant percentage of bone marrow transplanted patients with thalassemia. Factors responsible for short stature are previous iron overload, liver impairment, DFX treatment, and toxicity of chemotherapeutic agents. In some patients absence of pubertal development is due to gonadotropin insufficiency, probably secondary to previous iron overload; other patients exhibit hypergonadotrophic hypogonadism due to the toxic effect of chemotherapeutic agents on the gonads. Both groups need hormonal replacement therapy. These data support the need for vigilant follow-up of patients with thalassemia before and after transplantation, in order to treat endocrine dysfunctions at the appropriate age. PMID- 12729402 TI - Pubertal development in cystic fibrosis: an overview. AB - Cystic fibrosis (CF) is an autosomal recessive disease characterized by respiratory and intestinal insufficiencies. It has been reported in the literature that patients with CF show delayed growth and puberty and girls with CF achieve menarche at older age than normal females. Infertility, in both sexes, and menstrual dysfunction, in girls, are common in patients with CF. Previous data suggest that the degree of failure of growth and development is correlated with malnutrition and severity of progressing pulmonary disease. Despite improved nutrition and intensive treatment, patients with CF have delayed puberty and growth pubertal spurt. This maturational lag is accompanied by a significant delay in attaining pubertal levels of insulin-like growth factor-I (IGF-I), luteinizing hormone (LH), follicle-stimulating hormone (FSH) and sex steroid hormones. PMID- 12729403 TI - Pubertal growth and final height in children with chronic renal failure on conservative treatment. AB - In order to evaluate the pattern of pubertal growth in children with chronic renal failure (CRF) and its impact on the progression of renal impairment, the auxological data relating to ten boys and five girls with CRF were collected from the beginning to the end of puberty and compared with creatinine clearance (CCr) over the same period. The progression of renal impairment was evaluated as a regression line by plotting CCr by age before and after puberty. The magnitude (10.6 cm/year in males and 10.1 cm/year in females) and timing of peak height velocity (PHV) (13.6 and 11.3 years, respectively) were normal, as was the duration of puberty calculated as the interval between PHV and final height (FH). The slope of the mean regression line during the 5 years before and after PHV was respectively -1.1 +/- 4.2 and -3.7 +/- 3.2 (p <0.05, Mann-Whitney test for non parametric data). In conclusion, the boys showed a slight impairment in FH (-0.37 SDS), which was more severe in the girls who lost their growth potential in early childhood; the height increases and PHV-FH time intervals were similar to those observed in healthy children (p = NS for all of the considered variables). As the loss of renal function was greater in the post-PHV than prepubertal period, puberty per se may play a role in accelerating the decline in the glomerular filtration rate. This possibility deserves further investigation. PMID- 12729404 TI - Final height in congenital adrenal hyperplasia due to 21-hydroxylase deficiency: the Italian experience. AB - OBJECTIVE: To investigate the influence of target height (TH), gender, phenotype, glucocorticoid formulation and age at onset of treatment on final height (FH) in patients with 21-hydroxylase deficiency (21OHD). PATIENTS: Clinical data of 93 patients--46 simple virilizing (SV), 35 salt-wasting (SW) and 12 late onset (LO)- were collected in six pediatric endocrinology units in Italy. RESULTS: FH and TH were always below the mean height of the general population (mean FH, SDS: SW patients -1.3 +/- 1.2, SV patients -1.8 +/- 0.9, LO patients -1.7 +/- 1.1; mean TH, SDS: SW patients -0.6 +/- 0.8, SV patients -0.7 +/- 0.9, LO patients -1.4 +/- 1.3). FH was significantly below TH in patients with classic form (SW and SV, p <0.001), but not in LO patients. In classic form, TH seems to be related to FH, followed by age at onset of therapy and by steroid formulation, these variables explaining 30% of FH variance. CONCLUSIONS: In the classic form, substitutive therapy started before 21 months of age improved the long-term outcome. Lower TH in LO patients could be due to undiagnosed non-classic 21OHD in some of their parents. FH in LO patients seems not to benefit from corticosteroid therapy, even if late diagnosis may partly account for this result. PMID- 12729405 TI - Age at menarche in juvenile rheumatoid arthritis. AB - OBJECTIVE: To study the effect on the timing of menarche of the different subtypes of juvenile rheumatoid arthritis (JCA) and their therapies. METHODS: Auxological data (including the age at menarche of themselves and their mothers) were collected from 83 females regularly attending our rheumatological unit. Malnourished patients were excluded from the study. The patients were divided into groups on the basis of disease subtype and glucocorticoid (GC) therapy. RESULTS: The timing of menarche was later in all the patients in comparison with their mothers and normal Italian girls, and those who had taken GC had a later menarche and less pubertal growth than those who had not. The girls with systemic JCA had a later menarche than those with the polyarticular or pauciarticular subtypes. CONCLUSIONS: Our results show that pubertal development is compromised in girls with JCA, particularly if they have received GC therapy. PMID- 12729406 TI - Growth and pubertal disorders in neurofibromatosis type 1. AB - The first textbook of Pediatric Endocrinology in the early 1950s reported an association of neurofibromatosis type 1 (NF1) and precocious puberty (PP) and/or short stature. Recent studies have indicated that children with NF1 grow normally until puberty; thereafter height velocity and relative height (SDS or percentiles) decreases with respect to healthy peers, reaching a mean adult height close to the 25th percentile for the general population. Moreover, the percentage of patients with true short stature (<3rd percentile) increases from childhood (5%) to late puberty (20-30% in literature, 18% in our study), and final height is significantly below the genetic target and predicted adult height calculated just before or at the beginning of puberty. Finally, among the shortest patients (<10th percentile) there is a high incidence of severe complications, such as CNS tumors, huge plexiform neurofibromas and severe scoliosis. Precocious puberty is a frequent complication of NF1, and occurs mainly in association with optic pathway tumors (OPT); however, occasionally it has been reported in the absence of optic gliomas, probably with a similar incidence as in the general population. GnRH agonist therapy must be decided individually as in some patients further growth could be normal and/or treatment would not improve final height. In the presence of early pubertal signs, an OPT must be ruled out. In addition to PP, delayed puberty has been frequently reported in NF1. In a study of 123 girls with NF1, we found that the mean age at menarche (13.0 +/- 1.9 yr) was later than in their mothers (12.7 +/- 1.4 yr) and in the general population (12.4 +/- 1.2 yr; p <0.05), with a very high incidence of delayed menarche (>2 SD): 16% vs 6.8% (mothers) vs 3.4% (controls) (p <0.01). In conclusion, growth and puberty present unusual patterns in NF1, often with true pathological findings increasing medical and psychological problems. PMID- 12729407 TI - Pubertal development in patients with McCune-Albright syndrome or pseudohypoparathyroidism. AB - Gain or loss of function mutations of the GNAS1 gene lead to McCune-Albright syndrome (MAS) or pseudohypoparathyroidism Ia (PHP-Ia), respectively. Patients with MAS, caused by a post-zygotic missense mutation leading to constitutive activation of Gs alpha, suffer from gonadotropin-independent precocious puberty, and delayed or incomplete sexual development and reproductive dysfunction is found in women with PHP-Ia, an inherited disorder caused by deficient expression or function of the Gs alpha protein. In females with MAS, 50% developed precocious puberty by the age of 4 years, the remaining between 4 and 8 years. Peripheral precocious puberty is often atypical and characterized by alternate periods of rapid progression and regression of pubertal development; menstrual bleeding may occur before breast development. Ovarian cyst growth and regression is often described as a sign of ovarian follicle hyperactivation. Notwithstanding this clinical heterogeneity, a subset of patients with MAS develop relentlessly progressive precocious puberty ultimately resulting in premature epiphyseal fusion and reduced adult stature. Long-term information on reproductive function has been obtained in females: some patients had regular menses without ovarian cysts on pelvic US scan, whereas others were oligomenorrheic and showed recurrent ovarian cysts. In males with MAS, precocious puberty occurred in three patients between 4 and 9 years of age. In one patient, long-term follow-up demonstrated normal plasma testosterone and gonadotropin values at the age of 17 years. On testicular sonography, multiple hyperechogeneic spots were found in both testicles (snow-storm appearance). Female patients with PHP-Ia were oligomenorrheic or amenorrheic; more than half had delayed or incomplete sexual development, They were mildly hypoestrogenic with normal to slightly elevated serum gonadotropin levels. These clinical and biochemical findings indicate partial resistance of the theca and granulosa cells of the ovary to gonadotropins due to deficient Gs alpha activity. Responsiveness might be sufficient to promote some degree of follicular development and steroid secretion, but insufficient to induce ovulation PMID- 12729408 TI - Puberty in disorders of somatosexual differentiation. AB - Despite enormous advances in the understanding of the molecular mechanisms of sexual differentiation, the medical decisions made in patients with disorders of somatosexual differentiation are mostly lacking evidence-based principles. Recent critical discussions have focused on approaches to gender assignment in infants with ambiguous genitalia. These decisions must be based on the correct diagnosis to be able to predict the development during puberty and adulthood in the affected individual. Puberty in this process plays a pivotal role not only because of the physical changes induced by endogenous or supplementary sex steroids to enhance the sex of rearing, but it is also the time for sexual orientation and promotion of gender identity. In this review we focus not only on the molecular impact of sex steroid action, but also view current opinions and studies on the gender development of the intersex patient during puberty. PMID- 12729409 TI - Growth and puberty in Turner's syndrome. AB - Turner's syndrome is the commonest sex chromosome abnormality in females, resulting from the absence of an X chromosome or the presence of a structurally abnormal X chromosome. Short stature and ovarian failure are the most consistent clinical features and require specific and co-ordinated medical approaches in order to improve final height and well-being in adulthood. High doses of growth hormone (GH) are able to improve adult height in comparison with untreated patients. GH therapy should be started during childhood as soon as the growth curve declines below the 5th percentile and doses adjusted according to clinical response. Some studies reported that combined therapy with GH and an anabolic steroid--oxandrolone--is also beneficial at lower GH doses. Ovarian failure should be treated by appropriate substitutive estrogen therapy. Since estrogen administration may impair growth of patients with Turner's syndrome, the age at beginning of therapy should be individualized taking into consideration potential growth, the need for feminization, bone mineral density and the psychological well-being of each patient. Well co-ordinated endocrine interventions can permit better long-term outcome in adulthood. PMID- 12729410 TI - Growth and pubertal growth spurt in dysmorphic syndromes. AB - We describe peculiar aspects of growth and the pubertal growth spurt in dysmorphic syndromes, focusing our attention on the accelerated pubertal growth pattern in some dysmorphic syndromes leading to short stature (DSS). A normal pubertal growth spurt was observed in other dysmorphic syndromes leading to overgrowth (ODS). PMID- 12729412 TI - Acquisition of bone mass in normal individuals and in patients with growth hormone deficiency. AB - Children and adolescents with growth hormone (GH) deficiency (GHD) have reduced bone turnover and bone mass in comparison with normal individuals. GH replacement therapy stimulates bone turnover and improves bone mass during childhood and adolescence. At final height, patients with GHD treated with GH have normal mean values of bone mass, but some patients have reduced lumbar BMD (Z score <1) with an increased susceptibility to fractures. Children and adolescents with GHD have approximately a fourfold decreased fracture frequency during treatment in comparison with controls. The changes of bone mass during the transition to adulthood in patients with GHD who discontinued GH treatment after final height are not defined. An increase of lumbar BMD after final height has been reported until 2 years after discontinuation of GH treatment; this could be the result of a spontaneous progression towards peak bone mass or of a persisting effect of GH treatment on bone mass accumulation. GH has an important role in the accrual of bone mass during childhood and adolescence, but the impact of GHD on bone mass during the transiton to adulthood is unknown. PMID- 12729411 TI - Gonadal function and pubertal development after treatment of a childhood malignancy. AB - As survival rates for childhood cancer have improved, the importance of assessing gonadal dysfunction caused by alkylating agents and radiotherapy in children treated for cancer has increased. Infertility is the major long-term side effect of chemotherapy (CT) in males, whereas Leydig cell function is less affected. Our studies confirm that prepuberty does not protect the male gonad from the late effects of CT and that protocols less gonadal-lesive (such as ABVD regimens) should be preferred. Ovaries are less affected, but early depletion of follicles and premature menopause may occur. High-dose busulfan conditioning regimens cause ovarian failure in young females. The role of gonadal irradiation is discussed: high dosages (>2000 cGy) provoke sterility, impaired testosterone secretion in males and estradiol release in females. High dosage hypothalamic-pituitary irradiation causes delayed puberty and hypogonadism in males and females, whereas lower dosages may be associated with early puberty, particularly in females. PMID- 12729413 TI - Effects of thalassemia major on bone mineral density in late adolescence. AB - Management of thalassemia major has shown substantial clinical and prognostic improvement, suggesting the need for major attention to quality of life. We studied bone health in 25 patients (13 males, 12 females; 15-23 years old) affected by beta-thalassemia major. In all patients, bone mineral density (BMD), biochemical markers of bone and calcium metabolism (calcium, phosphate, magnesium, alkaline phosphatase, urinary calcium, 25-hydroxyvitamin D [25OH-D], 1,25-dihydroxyvitamin D [1,25(OH)2D], parathyroid hormone [PTH]), hematological parameters and gonadal steroids status were assessed and related to each other and to auxological parameters (chronological, statural and bone ages, height, weight, stage of puberty). BMD of the lumbar spine (L1-L4) (g/cm2) and expressed as Z-scores, was assessed by dual energy X-ray absorptiometry. PTH levels were low in seven patients (28%), and in the normal range in 18 (72%). 25OH-D serum levels were normal in 16 patients (64%) and low in nine (36%). 1,25(OH)2D values were reduced in 19 patients (76%) and normal in six (24%). Alkaline phosphatase correlated with bone age delay (r = 0.414; p = 0.039); no other statistically significant correlation was found. Mean BMD values in patients with thalassemia were significantly reduced in comparison with that of age- and sex-matched controls (Z-score: -2.8 +/- 2.0, p <0.001; -3.3 +/- 2.1 in males, and -2.2 +/- 1.9 in females). Twenty-one patients (84%) showed reduced BMD. Overall, BMD reduction was in the osteopenia range in five patients (20%) and in the osteoporosis range in 16 patients (64%). Our data indicate that low BMD is often present in patients with thalassemia, although recognized late, as in the present series. Early diagnosis should be done during childhood, in order to improve the quality of life in adulthood. PMID- 12729414 TI - Bone mineral density in long-term survivors of childhood cancer. AB - Due to developments in diagnostics and therapeutic methods, around two-thirds of the children diagnosed with cancer are cured and reach adulthood. More attention has been drawn to the adverse effects of cancer treatments including those on bone metabolism. Potentially harmful components of cancer therapies with regard to bone development are, e.g., corticosteroids, radiotherapy, and high doses of antimetabolites, such as methotrexate. In long-term survivors of childhood acute lymphoblastic leukemia and intracranial tumors, a history of cranial radiation has been associated with reduced bone mass. More controversy exists about the development of bone mass in long-term survivors of childhood cancer when cranial radiation has not been administered. Both normal as well as decreased bone mass and mineral density has been observed in this group of childhood cancer survivors. Long-term longitudinal data on bone mass development are required to further elucidate this question. PMID- 12729415 TI - Carcinogenic risk from hot particle exposures--has ICRP got it right? PMID- 12729416 TI - Carcinogenic risk of hot-particle exposures. AB - It has been suggested that spatially non-uniform radiation exposures, such as those from small radioactive particles ('hot particles'), may be very much more carcinogenic than when the same amount of energy is deposited uniformly throughout a tissue volume. This review provides a brief summary of in vivo and in vitro experimental findings, and human epidemiology data, which can be used to evaluate the veracity of this suggestion. Overall, this supports the contrary view and indicates that average dose, as advocated by the ICRP, is likely to provide a reasonable estimate of carcinogenic risk (within a factor of approximately +/- 3). There are few human data with which to address this issue. The limited data on lung cancer mortality following occupational inhalation of plutonium aerosols, and the incidence of liver cancer and leukaemia due to thorotrast administration for clinical diagnosis, do not appear to support a significant enhancement factor. Very few animal studies, including mainly lung and skin exposures, provide any indication of a hot-particle enhancement for carcinogenicity. Some recent in vitro malignant transformation experiments provide evidence foran enhanced cell transformation for hot-particle exposures but, properly interpreted, the effect is modest. Few studies extend below absorbed doses of approximately 0.1 Gy. PMID- 12729417 TI - Background radiation: natural and man-made. AB - A brief overview and comparison is given of dose rates arising from natural background radiation and the fallout from atmospheric testing of nuclear weapons. Although there are considerable spatial variations in exposure to natural background radiation, it is useful to give estimates of worldwide average overall exposures from the various components of that background. Cosmic-ray secondaries of low linear energy transfer (LET), mainly muons and photons, deliver about 280 microSv a(-1). Cosmic-ray neutrons deliver about another 100 microSv a(-1). These low- and high-LET exposures are relatively uniform to the whole body. The effective dose rate from cosmogenic radionuclides is dominated by the contribution of 12 microSv a(-1) from 14C. This is due to relatively uniform irradiation of all organs and tissues from low-energy beta particles. Primordial radionuclides and their progeny (principally the 238U and 232Th series, and 40K) contribute about 480 microSv a(-1) of effective dose by external irradiation. This is relatively uniform photon irradiation of the whole body. Internally incorporated 40K contributes a further 165 microSv a(-1) of effective dose in adults, mainly from beta particles, but with a significant gamma component. Equivalent doses from 40K are somewhat higher in muscle than other soft tissues, but the distinction is less than a factor of three. Uranium and thorium series radionuclides give rise to an average effective dose rate of around 120 microSv a(-1). This includes a major alpha particle component, and exposures of radiosensitive tissues in lung, liver, kidney and the skeleton are recognised as important contributors to effective dose. Overall, these various sources give a worldwide average effective dose rate of about 1160 microSv a(-1). Exposure to 222Rn, 220Rn and their short-lived progeny has to be considered separately. This is very variable both within and between countries. For 222Rn and its progeny, a worldwide average effective dose rate is about 1105 microSv a(-1). For 220Rn and its progeny, the corresponding value is 91 microSv a(-1). In both cases, the effective dose is mainly due to a particle irradiation of the bronchial tissues of the lungs. Overall, the worldwide average effective dose rate from natural background is about 2400 microSv a(-1) or 2.4 mSv a(-1). For comparison, worldwide average effective dose rates from weapons fallout peaked at 113 microSv a(-1) (about 5% of natural background) in 1963 and have since fallen to about 5.5 microSv a(-1) (about 0.2% of natural background). These values perhaps serve to emphasise that even gross insults to the natural environment from anthropogenic releases of radioactive materials are likely to be of limited significance when set in the context of the ambient radioactive environment within which all organisms, including humans, have developed. PMID- 12729418 TI - Review of the radioactive waste management system in Nigeria. AB - The management of radioactive waste in Nigeria from early 1960 to date is reviewed. As in many developing countries, waste management in Nigeria has been shown to be ineffective. The factors that are responsible for this ineffectiveness are identified and discussed. The steps being taken by and the opportunities available to the newly established Nigerian Nuclear Regulatory Authority towards addressing this problem of ineffectiveness are discussed. The efforts of this newly set up body towards managing the resultant radioactive wastes that will be generated during the use of a reactor and an accelerator that will soon be commissioned in Nigeria are also mentioned. Likely ways of further addressing the problems militating against waste management in developing countries are suggested. PMID- 12729420 TI - Investigation using an advanced extremity gamma instrumentation system of options for shielding the hand during the preparation and injection of radiopharmaceuticals. AB - Staff preparing and injecting radiopharmaceuticals in hospitals may receive significant radiation doses to their hands. These doses may be high enough to warrant that they be classified as radiation workers. The influence of local shielding on finger doses has been investigated. Staff preparing radioactive liquids in a radionuclide dispensary and drawing up and injecting radiopharmaceuticals in a nuclear medicine department have been studied. Measurements have been recorded with an electronic extremity dose monitor, an advanced extremity gamma instrumentation system (AEGIS), worn near to the finger tip. The electronic dosimeter allows the pattern of doses received during different procedures to be determined. Doses received for individual manipulations during many routine sessions have been recorded for different staff members. Dose distributions around shielded vials and syringes have also been measured using AEGIS. In the radionuclide dispensary the vials from which radioactive liquids are dispensed are held in tungsten shields, whereas in nuclear medicine simple lead pots are used. Syringe shields are employed for some parts of dispensing and patient injections. Data on dose distributions have been used in interpretation of results from monitoring. Use of syringe shields during dispensing reduced the finger dose by 75-85%. The peaks in dose rate were 60% lower, and periods of exposure to high dose rates were reduced in length by a third because of the restriction in the region of high dose rate. The extremity doses to staff dispensing and injecting radiopharmaceuticals in nuclear medicine were of similar magnitude. Doses received during dispensing varied from 10 to 555 microGy depending upon whether the vial containing the radiopharmaceutical was directly handled or not. Dose received from individual injections varied from 1 to 150 microGy depending on the degree of difficulty experienced during the injection. PMID- 12729419 TI - A contribution to the linear no-threshold discussion. AB - The paper approaches the linear no-threshold (LNT) hypothesis, currently used as the basis for recommendations in radiological protection, from the point of view of the radiation mechanism. All considerations of the validity of the LNT hypothesis based on experiment or epidemiology are dismissed because of the impossibility of deriving statistically significant data at very low doses. Instead, the LNT hypothesis is assessed from a consideration of the mechanism of radiation action. The DNA double-strand break is proposed to be the crucial radiation-induced molecular lesion. A trace is made using a series of correlations that link the DNA double-strand break to effects at the cellular level and these cellular effects are linked to the induction of cancer. Multistep modelling of carcinogenesis is used to take the link through to a consideration of radiation risk. It is concluded that, from the point of view of radiation mechanism, at very low doses the LNT hypothesis of radiation action is valid, that is, the risk function has a positive slope from zero dose. PMID- 12729421 TI - Predicting the 137Cs ingestion dose from marine fish consumption in Hong Kong after an accidental release. AB - Caesium-137 is one of the more important radionuclides released during a nuclear power plant accident. Based on a mathematical model developed earlier by the present authors for estimating 137Cs ingestion dose from consumption of marine fish in Hong Kong, doses due to an accidental release into Daya Bay from the Guangdong Nuclear Power Station are predicted. Assuming a release of 30000 TBq of 137Cs, results show that the cumulative dose rises rapidly during the first five years and then gradually levels off. In the first and 50th years after discharge, doses to an average Hong Kong citizen would cumulate to 17 and 38 microSv respectively, which are dominated by the contribution from fish cultured in Hong Kong waters. The first year dose to members of the critical group of local fish farmers is estimated to be 1600 microSv, accumulating to 3600 microSv in the 50th year. Apart from this small number of critical group members whose doses may need some monitoring, the doses to the population at large are considered small compared with an annual dose of 175 microSv from ingestion of natural radionuclides in food and drinks in Hong Kong. PMID- 12729422 TI - Radioactivity in food and the environment: calculations of UK radiation doses using integrated methods. PMID- 12729423 TI - Duration of antibiotic therapy for early Lyme disease. A randomized, double blind, placebo-controlled trial. AB - BACKGROUND: Treatment of patients with early Lyme disease has trended toward longer duration despite the absence of supporting clinical trials. OBJECTIVE: To evaluate different durations of oral doxycycline treatment and the combination of oral doxycycline and a single intravenous dose of ceftriaxone for treatment of patients with early Lyme disease. DESIGN: Randomized, double-blind, placebo controlled trial. SETTING: Single-center university hospital. PATIENTS: 180 patients with erythema migrans. INTERVENTION: Ten days of oral doxycycline, with or without a single intravenous dose of ceftriaxone, or 20 days of oral doxycycline. MEASUREMENTS: Outcome was based on clinical observations and neurocognitive testing. Efficacy was assessed at 20 days, 3 months, 12 months, and 30 months. RESULTS: At all time points, the complete response rate was similar for the three treatment groups in both on-study and intention-to-treat analyses. In the on-study analysis, the complete response rate at 30 months was 83.9% in the 20-day doxycycline group, 90.3% in the 10-day doxycycline group, and 86.5% in the doxycycline-ceftriaxone group (P > 0.2). The only patient with treatment failure (10-day doxycycline group) developed meningitis on day 18. There were no significant differences in the results of neurocognitive testing among the three treatment groups and a separate control group without Lyme disease. Diarrhea occurred significantly more often in the doxycycline ceftriaxone group (35%) than in either of the other two groups (P < 0.001). CONCLUSIONS: Extending treatment with doxycycline from 10 to 20 days or adding one dose of ceftriaxone to the beginning of a 10-day course of doxycycline did not enhance therapeutic efficacy in patients with erythema migrans. Regardless of regimen, objective evidence of treatment failure was extremely rare. PMID- 12729424 TI - Intravenous and oral itraconazole versus intravenous and oral fluconazole for long-term antifungal prophylaxis in allogeneic hematopoietic stem-cell transplant recipients. A multicenter, randomized trial. AB - BACKGROUND: Allogeneic hematopoietic stem-cell transplant recipients often receive fluconazole or an amphotericin B preparation for antifungal prophylaxis. Because of concerns about fungal resistance with fluconazole and toxicity with amphotericin B, alternative prophylactic regimens have become necessary. OBJECTIVE: To compare the efficacy and safety of intravenous and oral itraconazole with the efficacy and safety of intravenous and oral fluconazole for long-term prophylaxis of fungal infections. DESIGN: Open-label, multicenter, randomized trial. SETTING: Five transplantation centers in the United States. PATIENTS: 140 patients undergoing allogeneic hematopoietic stem-cell transplantation. INTERVENTION: Itraconazole (200 mg intravenously every 12 hours for 2 days followed by 200 mg intravenously every 24 hours or a 200-mg oral solution every 12 hours) or fluconazole (400 mg intravenously or orally every 24 hours) from day 1 until day 100 after transplantation. MEASUREMENTS: Proven invasive or superficial fungal infection, drug-related side effects, mortality from fungal infection, and overall mortality. RESULTS: Proven invasive fungal infections occurred in 6 of 71 itraconazole recipients (9%) and in 17 of 67 fluconazole recipients (25%) during the first 180 days after transplantation (difference, -16 percentage points [95% CI, -29.2 to -4.7 percentage points]; P = 0.01). Superficial fungal infections occurred in 3 of 71 itraconazole recipients (4%) and in 2 of 67 fluconazole recipients (3%). In a multivariable analysis using factors known to affect the risk for invasive fungal infection after hematopoietic stem-cell transplantation, prophylaxis with itraconazole was still associated with fewer invasive fungal infections (odds ratio, 0.300 [CI, 0.111 to 0.814]; P = 0.02) caused by either yeasts or molds. More fungal pathogens were found to be resistant to fluconazole than to itraconazole. Except for more frequent gastrointestinal side effects (nausea, vomiting, diarrhea, or abdominal pain) in patients given itraconazole (24% vs. 9%; difference, 15 percentage points [CI, 2.9 to 27.0 percentage points]; P = 0.02), both itraconazole and fluconazole were well tolerated. The overall mortality rate was similar in each group (32 of 71 patients in the itraconazole group [45%] vs. 28 of 67 patients in the fluconazole group [42%]; difference, 3 percentage points [CI, -13.2 to 19.8 percentage points]; P > 0.2), but fewer deaths were related to fungal infection in patients given itraconazole (6 of 71 [9%]) than in patients given fluconazole (12 of 67 [18%]) (difference, 9 percentage points [CI, -20.6 to 1.8 percentage points]; P = 0.13). CONCLUSION: Itraconazole is more effective than fluconazole for long-term prophylaxis of invasive fungal infections after allogeneic hematopoietic stem-cell transplantation. Except for gastrointestinal side effects, itraconazole is well tolerated. PMID- 12729425 TI - Comparison of 10-mg and 5-mg warfarin initiation nomograms together with low molecular-weight heparin for outpatient treatment of acute venous thromboembolism. A randomized, double-blind, controlled trial. AB - BACKGROUND: The optimal means of achieving therapeutic oral anticoagulation in the outpatient setting has not been determined. OBJECTIVE: To compare a 10-mg dosing nomogram with a 5-mg nomogram that has been suggested to be sufficient for warfarin initiation. DESIGN: Randomized, controlled clinical trial. SETTING: Outpatient venous thromboembolism services of four tertiary care hospitals. PATIENTS: 201 of 210 consecutive patients with objectively confirmed diagnoses of acute venous thromboembolism. INTERVENTION: All patients were treated with subcutaneous low-molecular-weight heparin for a minimum of 5 days until a therapeutic international normalized ratio (INR) was achieved. Patients were randomly assigned to initially receive a 10-mg or 5-mg dose of warfarin. MEASUREMENTS: The primary end point was time in days to therapeutic INR. Secondary end points were the proportion of patients who had achieved a therapeutic INR by day 5, the total number of INR assessments, the number of INR measurements greater than 5.0, incidence of recurrent venous thromboembolism and major bleeding, and survival. RESULTS: 210 consecutive patients met the inclusion criteria. Of these, 9 were excluded and 201 were randomly assigned to study groups (104 to the 10-mg group and 97 to the 5-mg group). Demographic characteristics of both groups were similar. Patients in the 10-mg group achieved therapeutic INR 1.4 days earlier than patients in the 5-mg group (P < 0.001). Eighty-three percent of patients in the 10-mg group achieved a therapeutic INR by day 5 versus 46% in the 5-mg group (P < 0.001). Fewer INR assessments were performed in the 10-mg group than in the 5-mg group (8.1 vs. 9.1; P = 0.04). There were no significant differences between the two groups in recurrent events, major bleeding, survival, and number of INR measurements greater than 5.0. CONCLUSION: The 10-mg warfarin initiation nomogram is superior to the 5-mg nomogram because it allows more rapid achievement of a therapeutic INR. PMID- 12729426 TI - Suboptimal monitoring and dosing of unfractionated heparin in comparative studies with low-molecular-weight heparin. AB - BACKGROUND: Site-specific validation of the activated partial thromboplastin time (aPTT) therapeutic range is required to ensure administration of the optimal dose of unfractionated heparin. Therapeutic ranges of 1.5 to 2.5 times the control value are subtherapeutic for most modern aPTT reagents. PURPOSE: To audit the appropriateness of aPTT monitoring in clinical trials comparing unfractionated heparin and low-molecular-weight heparin in patients with venous thromboembolism. DATA SOURCES: Search of PubMed database from 1984 to 2001. STUDY SELECTION: Randomized, controlled trials that compared unfractionated and low-molecular weight heparin for the treatment of venous thromboembolism. DATA EXTRACTION: Use of unvalidated and potentially suboptimal therapeutic ranges for aPTT in patients assigned to receive unfractionated heparin. DATA SYNTHESIS: Fifteen studies met inclusion criteria. Only 3 studies used a validated aPTT therapeutic range, and 11 studies used a range that included aPTT values 1.5 times the control value. Ten studies reported unfractionated heparin doses, and 7 of these documented a reduction to less than 30 000 U/d in response to aPTT results. CONCLUSIONS: Most studies monitored unfractionated heparin inappropriately. This shortcoming could be responsible for the reduced efficacy of unfractionated heparin in clinical trials. PMID- 12729427 TI - Cost-effectiveness of alternative management strategies for patients with solitary pulmonary nodules. AB - BACKGROUND: Positron emission tomography (PET) with 18-fluorodeoxyglucose (FDG) is a potentially useful but expensive test to diagnose solitary pulmonary nodules. OBJECTIVE: To evaluate the cost-effectiveness of strategies for pulmonary nodule diagnosis and to specifically compare strategies that did and did not include FDG-PET. DESIGN: Decision model. DATA SOURCES: Accuracy and complications of diagnostic tests were estimated by using meta-analysis and literature review. Modeled survival was based on data from a large tumor registry. Cost estimates were derived from Medicare reimbursement and other sources. TARGET POPULATION: All adult patients with a new, noncalcified pulmonary nodule seen on chest radiograph. TIME HORIZON: Patient lifetime. PERSPECTIVE: Societal. INTERVENTION: 40 clinically plausible combinations of 5 diagnostic interventions, including computed tomography, FDG-PET, transthoracic needle biopsy, surgery, and watchful waiting. OUTCOME MEASURES: Costs, quality-adjusted life-years (QALYs), and incremental cost-effectiveness ratios. RESULTS OF BASE CASE ANALYSIS: The cost-effectiveness of strategies depended critically on the pretest probability of malignancy. For patients with low pretest probability (26%), strategies that used FDG-PET selectively when computed tomography results were possibly malignant cost as little as 20 000 dollars per QALY gained. For patients with high pretest probability (79%), strategies that used FDG-PET selectively when computed tomography results were benign cost as little as 16 000 dollars per QALY gained. For patients with intermediate pretest probability (55%), FDG-PET strategies cost more than 220 000 dollars per QALY gained because they were more costly but only marginally more effective than computed tomography based strategies. RESULTS OF SENSITIVITY ANALYSIS: The choice of strategy also depended on the risk for surgical complications, the probability of nondiagnostic needle biopsy, the sensitivity of computed tomography, and patient preferences for time spent in watchful waiting. In probabilistic sensitivity analysis, FDG PET strategies were cost saving or cost less than 100 000 dollars per QALY gained in 76.7%, 24.4%, and 99.9% of computer simulations for patients with low, intermediate, and high pretest probability, respectively. CONCLUSIONS: FDG-PET should be used selectively when pretest probability and computed tomography findings are discordant or in patients with intermediate pretest probability who are at high risk for surgical complications. In most other circumstances, computed tomography-based strategies result in similar quality-adjusted life years and lower costs. PMID- 12729428 TI - Eponyms and the diagnosis of aortic regurgitation: what says the evidence? AB - BACKGROUND: Chronic aortic regurgitation can lead to significant morbidity and mortality. For more than a century, numerous eponymous signs of aortic regurgitation have been described in textbooks and the literature. PURPOSE: To compare current textbook content with the peer-reviewed literature on the eponymous signs of aortic regurgitation and to assess the role of these signs in clinical practice. DATA SOURCES: 11 textbooks, MEDLINE (1966 through October 2002), and bibliographies of textbooks and relevant papers. STUDY SELECTION: English-language reports that were related to the properties of a sign on physical examination, incorporated more than 10 adults, and did not involve prosthetic heart valves or acute aortic regurgitation. DATA EXTRACTION: Three investigators independently analyzed relevant textbook extracts and 27 reports, using predetermined qualitative review criteria. Data relating to diagnostic accuracy and properties of the index test were also extracted. DATA SYNTHESIS: Twelve eponymous signs were described as having varying degrees of importance by textbook authors. Only the Austin Flint murmur, the Corrigan pulse, the Duroziez sign, and the Hill sign had sufficient original literature for detailed review. Most reports were low quality, with varying sensitivities for all signs. Except for the Hill sign, specificity tended to be poor. Evidence for the Hill sign also suggested a correlation between the popliteal-brachial gradient and aortic regurgitation severity. CONCLUSIONS: Prominent textbook support of the eponymous signs of aortic regurgitation is not matched by the literature. Clinicians and educators should update and improve the evidence for these signs to ensure their relevance in current medical practice. PMID- 12729429 TI - Resolution of futility by due process: early experience with the Texas Advance Directives Act. AB - Every U.S. state has developed legal rules to address end-of-life decision making. No law to date has effectively dealt with medical futility--an issue that has engendered significant debate in the medical and legal literature, many court cases, and a formal opinion from the American Medical Association's Council on Ethical and Judicial Affairs. In 1999, Texas was the first state to adopt a law regulating end-of-life decisions, providing a legislatively sanctioned, extrajudicial, due process mechanism for resolving medical futility disputes and other end-of-life ethical disagreements. After 2 years of practical experience with this law, data collected at a large tertiary care teaching hospital strongly suggest that the law represents a first step toward practical resolution of this controversial area of modern health care. As such, the law may be of interest to practitioners, patients, and legislators elsewhere. PMID- 12729430 TI - Training and competency evaluation for interpretation of 12-lead electrocardiograms: recommendations from the American College of Physicians. AB - This paper is part 1 of a 2-part series on interpretation of 12-lead resting electrocardiograms (ECGs). Part 1 is a position paper that presents recommendations for initial competency, competency assessment, and maintenance of competency on ECG interpretation, as well as recommendations for the role of computer-assisted ECG interpretation. Part 2 is a systematic review of detailed supporting evidence for the recommendations. Despite several earlier consensus based recommendations on ECG interpretation, substantive evidence on the training needed to obtain and maintain ECG interpretation skills is not available. Some studies show that noncardiologist physicians have more ECG interpretation errors than do cardiologists, but the rate of adverse patient outcomes from ECG interpretation errors is low. Computers may decrease the time needed to interpret ECGs and can reduce ECG interpretation errors. However, they have shown less accuracy than physician interpreters and must be relied on only as an adjunct interpretation tool for a trained provider. Interpretation of ECGs varies greatly, even among expert electrocardiographers. Noncardiologists seem to be more influenced by patient history in interpreting ECGs than are cardiologists. Cardiologists also perform better than other specialists on standardized ECG examinations when minimal patient history is provided. Pending more definitive research, residency training in internal medicine with Advanced Cardiac Life Support instruction should continue to be sufficient for bedside interpretation of resting 12-lead ECGs in routine and emergency situations. Additional experience or training in ECG interpretation when the patient's clinical condition is unknown may be useful but requires further study. PMID- 12729431 TI - Competency in interpretation of 12-lead electrocardiograms: a summary and appraisal of published evidence. AB - BACKGROUND: There have been many proposals for objective standards designed to optimize training, testing, and maintaining competency in interpretation of electrocardiograms (ECGs). However, most of these recommendations are consensus based and are not derived from clinical trials that include patient outcomes. PURPOSE: To critically review the available data on training, accuracy, and outcomes of computer and physician interpretation of 12-lead resting ECGs. DATA SOURCES: English-language articles were retrieved by searching MEDLINE (1966 to 2002), EMBASE (1974 to 2002), and the Cochrane Controlled Trials Register (1975 2002). The references in articles selected for analysis were also reviewed for relevance. STUDY SELECTION: All articles on training, accuracy, and outcomes of ECG interpretations were analyzed. DATA EXTRACTION: Study design and results were summarized in evidence tables. Information on physician interpretation compared to a "gold standard," typically a consensus panel of expert electrocardiographers, was extracted. The clinical context of and outcomes related to the ECG interpretation were obtained whenever possible. DATA SYNTHESIS: Physicians of all specialties and levels of training, as well as computer software for interpreting ECGs, frequently made errors in interpreting ECGs when compared to expert electrocardiographers. There was also substantial disagreement on interpretations among cardiologists. Adverse patient outcomes occurred infrequently when ECGs were incorrectly interpreted. CONCLUSIONS: There is no evidence-based minimum number of ECG interpretations that is ideal for attaining or maintaining competency in ECG interpretation skills. Further research is needed to clarify the optimal way to build and maintain ECG interpretation skills based on patient outcomes. PMID- 12729432 TI - Duration of antibiotic therapy for Lyme disease. PMID- 12729433 TI - Dark rounds. PMID- 12729434 TI - Stuck. PMID- 12729435 TI - Screening for depression in adults. PMID- 12729436 TI - Screening for depression in adults. PMID- 12729438 TI - Screening for breast cancer. PMID- 12729439 TI - Screening for breast cancer. PMID- 12729441 TI - The mammography dilemma. PMID- 12729444 TI - The mammography dilemma. PMID- 12729445 TI - What do doctors find meaningful about their work? PMID- 12729446 TI - Summaries for patients. Duration of antibiotic treatment for early Lyme disease. PMID- 12729447 TI - Summaries for patients. What is the most effective blood thinner for treating patients with blood clots in the veins? PMID- 12729448 TI - Summaries for patients. A comparison of two methods of starting the anticoagulant drug warfarin. PMID- 12729449 TI - Summaries for patients. Preventing fungal infections in patients with stem-cell transplants. PMID- 12729450 TI - Quality grand rounds. PMID- 12729451 TI - Disparities in health and health care: focusing efforts to eliminate unequal burdens. AB - Disparities in health and health care have been around for more than two centuries. Evidence suggests that health disparities in ethnic and racial minorities continue to be problematic, with little progress made to eliminate them over time. Ethnic and racial disparities exist for multiple and complex reasons. However, new solutions are needed to resolve some of these old problems. Framing the debate and discussion around the distinctiveness related to disparities in health and health care is a necessary beginning in eliminating unequal burdens in health status. Focusing efforts to eliminate unequal burdens can strengthen existing solutions and policy formation related to this issue. This article defines disparities in health and health care, describes current health disparities impacting ethnic/racial groups, reviews historical factors associated with existing disparities in ethnic/racial groups, and concludes with challenges and solutions to alleviate these disparities. PMID- 12729452 TI - The many faces of diversity: overview and summary. PMID- 12729453 TI - Many faces: addressing diversity in health care. AB - DIVERSITY: The changing demographics and economics of our growing multicultural world, and the long-standing disparities in the health status of people from culturally diverse backgrounds has challenged health care providers and organizations to consider cultural diversity as a priority. The purpose of this article is to present a model that will be helpful in providing culturally competent care. The concept of cultural competence is discussed, "The Process of Cultural Competence in the Delivery of Healthcare Services Model" is described, and a mnemonic to guide in providing culturally competent care is presented. PMID- 12729454 TI - Race consciousness and the health of African Americans. AB - The historical experience of African Americans in our country has been shaped by the institution of slavery, dehumanization of blacks, segregation, pursuit of civil rights, and racism in contemporary American society. Disparities in health care provide compelling evidence that issues of race or skin color for the descendants of slaves and other ethnic minorities persist in the 21st century. Nurses providing care for African Americans must bridge the racial divide and incorporate culturally relevant content in the health history. As an integral aspect of their professional growth as culturally competent health care providers, they must incorporate the idea of "race consciousness" which is described as an awareness of the historical journey of the group, knowledge of disparities in health care for the people, and a self appraisal of one's attitudes and biases toward the group. PMID- 12729455 TI - Ethics and terrorism: September 11, 2001 and its aftermath. PMID- 12729456 TI - Quality of health information on the Web: where are we now? PMID- 12729457 TI - What does the nurse reinvestment act mean to you? AB - During 2002, the 107th Congress passed landmark legislation, the Nurse Reinvestment Act, P.L. 107-205. This article discusses the specific provisions of P.L. 107-205 within the context of the contemporary literature and the experience of the nursing shortage. The authors ask nurses to examine what the Nurse Reinvestment Act means for their career development. In laying out the anatomy of the Nurse Reinvestment Act, title by title, and section by section, the article presents the Congressional plan for addressing the two faces of the shortage: Nurse Recruitment, Title I, and Nurse Retention, Title II. PMID- 12729458 TI - Interweaving policy and diversity. AB - Nurses too often consider that direct patient care is the only focus of nursing. However, the role of the nurse is broader than that of direct care and encompasses many components, all of which must be integrated into the self of the nurse. Political activity and policy formation are two areas of nursing that are misunderstood and are not adopted enthusiastically by many nurses. This article examines the whole policy process that includes agenda setting; government response via laws, regulations and programs; policy/program implementation; and policy/program evaluation. The process is then linked to the concept of diversity to illustrate the need, ease and opportunity for nurse involvement. PMID- 12729459 TI - Ethics and the brave new world of e-Health. PMID- 12729460 TI - Extracellular ATP stimulates the early growth response protein 1 (Egr-1) via a protein kinase C-dependent pathway in the human osteoblastic HOBIT cell line. AB - Extracellular nucleotides exert an important role in controlling cell physiology by activating intracellular signalling cascades. Osteoblast HOBIT cells express P2Y(1) and P2Y(2) G-protein-coupled receptors, and respond to extracellular ATP by increasing cytosolic calcium concentrations. Early growth response protein 1 (Egr-1) is a C(2)H(2)-zinc-finger-containing transcriptional regulator responsible for the activation of several genes involved in the control of cell proliferation and apoptosis, and is thought to have a central role in osteoblast biology. We show that ATP treatment of HOBIT cells increases Egr-1 protein levels and binding activity via a mechanism involving a Ca(2+)-independent protein kinase C isoform. Moreover, hypotonic stress and increased medium turbulence, by inducing ATP release, result in a similar effect on Egr-1. Increased levels of Egr-1 protein expression and activity are achieved at very early times after stimulation (5 min), possibly accounting for a rapid way for changing the osteoblast gene-expression profile. A target gene for Egr-1 that is fundamental in osteoblast physiology, COL1A2, is up-regulated by ATP stimulation of HOBIT cells in a timescale that is compatible with that of Egr-1 activation. PMID- 12729462 TI - Nitrogen uptake in riparian plant communities across a sharp ecological boundary of salmon density. AB - BACKGROUND: Recent studies of anadromous salmon (Oncorhynchus spp.) on the Pacific Coast of North America indicate an important and previously unrecognized role of salmonid nutrients to terrestrial biota. However, the extent of this uptake by primary producers and consumers and the influences on community structure remain poorly described. We examine here the contribution of salmon nutrients to multiple taxa of riparian vegetation (Blechnum spicant, Menziesii ferruginea, Oplopanax horridus, Rubus spectabilis, Vaccinium alaskaense, V. parvifolium, Tsuga heterophylla) and measure foliar delta15N, total %N and plant community structure at two geographically separated watersheds in coastal British Columbia. To reduce potentially confounding effects of precipitation, substrate and other abiotic variables, we made comparisons across a sharp ecological boundary of salmon density that resulted from a waterfall barrier to salmon migration. RESULTS: delta15N and %N in foliage, and %cover of soil nitrogen indicators differed across the waterfall barrier to salmon at each watershed. delta15N values were enriched by 1.4 per thousand to 9.0 per thousand below the falls depending on species and watershed, providing a relative contribution of marine-derived nitrogen (MDN) to vegetation of 10% to 60%. %N in foliar tissues was slightly higher below the falls, with the majority of variance occurring between vegetation species. Community structure also differed with higher incidence of nitrogen-rich soil indicator species below the waterfalls. CONCLUSIONS: Measures of delta15N, %N and vegetation cover indicate a consistent difference in the riparian community across a sharp ecological boundary of salmon density. The additional N source that salmon provide to nitrogen-limited habitats appears to have significant impacts on the N budget of riparian vegetation, which may increase primary productivity, and result in community shifts between sites with and without salmon access. This, in turn, may have cascading ecosystem effects in forests adjacent to salmon streams. PMID- 12729461 TI - Fractalkine (CX3CL1) stimulated by nuclear factor kappaB (NF-kappaB)-dependent inflammatory signals induces aortic smooth muscle cell proliferation through an autocrine pathway. AB - Fractalkine (also known as CX3CL1), a CX3C chemokine, activates and attracts monocytes/macrophages to the site of injury/inflammation. It binds to CX3C receptor 1 (CX3CR1), a pertussis toxin-sensitive G-protein-coupled receptor. In smooth muscle cells (SMCs), fractalkine is induced by proinflammatory cytokines [tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma)], which may mediate monocyte adhesion to SMCs. However, the mechanisms underlying its induction are unknown. In addition, it is unlear whether SMCs express CX3CR1. TNF-alpha activated nuclear factor kappaB (NF-kappaB) and induced fractalkine and CX3CR1 expression in a time-dependent manner in rat aortic SMCs. Transient transfections with dominant-negative (dn) inhibitory kappaB (IkappaB)-alpha, dnIkappaB-beta, dnIkappaB kinase (IKK)-gamma, kinase-dead (kd) NF-kappaB-inducing kinase (NIK) and kdIKK-beta, or pretreatment with wortmannin, Akt inhibitor, pyrrolidinecarbodithioc acid ammonium salt ('PDTC') or MG-132, significantly attenuated TNF-alpha-induced fractalkine and CX3CR1 expression. Furthermore, expression of dn TNF-alpha-receptor-associated factor 2 (TRAF2), but not dnTRAF6, inhibited TNF-alpha signal transduction. Pretreatment with pertussis toxin or neutralizing anti-CX3CR1 antibodies attenuated TNF-alpha-induced fractalkine expression, indicating that fractalkine autoregulation plays a role in TNF-alpha induced sustained fractalkine expression. Fractalkine induced its own expression, via pertussis toxin-sensitive G-proteins, phosphoinositide 3-kinase (PI 3 kinase), phosphoinositide-dependent kinase 1 (PDK1), Akt, NIK, IKK and NF-kappaB activation, and induced SMC cell-cell adhesion and cellular proliferation. Taken together, our results demonstrate that TNF-alpha induces the expression of fractalkine and CX3CR1 in rat aortic SMCs and that this induction is mediated by NF-kappaB activation. We also show that fractalkine induces its own expression, which is mediated by the PI 3-kinase/PDK1/Akt/NIK/IKK/NF-kappaB signalling pathway. More importantly, fractalkine increased cell-cell adhesion and aortic SMC proliferation, indicating a role in initiation and progression of atherosclerotic vascular disease. PMID- 12729464 TI - Comparison of two DNA targets for the diagnosis of Toxoplasmosis by real-time PCR using fluorescence resonance energy transfer hybridization probes. AB - BACKGROUND: Toxoplasmosis is an infectious disease caused by the parasitic protozoan Toxoplasma gondii. It is endemic worldwide and, depending on the geographic location, 15 to 85% of the human population are asymptomatically infected. Routine diagnosis is based on serology. The parasite has emerged as a major opportunistic pathogen for immunocompromised patients, in whom it can cause life-threatening disease. Moreover, when a pregnant woman develops a primary Toxoplasma gondii infection, the parasite may be transmitted to the fetus and cause serious damage. For these two subpopulations, a rapid and accurate diagnosis is required to initiate treatment. Serological diagnosis of active infection is unreliable because reactivation is not always accompanied by changes in antibody levels, and the presence of IgM does not necessarily indicate recent infection. Application of quantitative PCR has evolved as a sensitive, specific, and rapid method for the detection of Toxoplasma gondii DNA in amniotic fluid, blood, tissue samples, and cerebrospinal fluid. METHODS: Two separate, real-time fluorescence PCR assays were designed and evaluated with clinical samples. The first, targeting the 35-fold repeated B1 gene, and a second, targeting a newly described multicopy genomic fragment of Toxoplasma gondii. Amplicons of different intragenic copies were analyzed for sequence heterogeneity. RESULTS: Comparative LightCycler experiments were conducted with a dilution series of Toxoplasma gondii genomic DNA, 5 reference strains, and 51 Toxoplasma gondii-positive amniotic fluid samples revealing a 10 to 100-fold higher sensitivity for the PCR assay targeting the newly described 529-bp repeat element of Toxoplasma gondii. CONCLUSION: We have developed a quantitative LightCycler PCR protocol which offer rapid cycling with real-time, sequence-specific detection of amplicons. Results of quantitative PCR demonstrate that the 529-bp repeat element is repeated more than 300-fold in the genome of Toxoplasma gondii. Since individual intragenic copies of the target are conserved on sequence level, the high copy number leads to an ultimate level of analytical sensitivity in routine practice. This newly described 529-bp repeat element should be preferred to less repeated or more divergent target sequences in order to improve the sensitivity of PCR tests for the diagnosis of toxoplasmosis. PMID- 12729463 TI - Explaining the de-prioritization of primary prevention: physicians' perceptions of their role in the delivery of primary care. AB - BACKGROUND: While physicians are key to primary preventive care, their delivery rate is sub-optimal. Assessment of physician beliefs is integral to understanding current behavior and the conceptualization of strategies to increase delivery. METHODS: A focus group with regional primary care physician (PCP) Opinion Leaders was conducted as a formative step towards regional assessment of attitudes and barriers regarding preventive care delivery in primary care. Following the PRECEDE-PROCEED model, the focus group aim was to identify conceptual themes that characterize PCP beliefs and practices regarding preventive care. Seven male and five female PCPs (family medicine, internal medicine) participated in the audiotaped discussion of their perceptions and behaviors in delivery of primary preventive care. The transcribed audiotape was qualitatively analyzed using grounded theory methodology. RESULTS: The PCPs' own perceived role in daily practice was a significant barrier to primary preventive care. The prevailing PCP model was the "one-stop-shop" physician who could provide anything from primary to tertiary care, but whose provision was dominated by the delivery of immediate diagnoses and treatments, namely secondary care. CONCLUSIONS: The secondary tertiary prevention PCP model sustained the expectation of immediacy of corrective action, cure, and satisfaction sought by patients and physicians alike, and, thereby, de-prioritized primary prevention in practice. Multiple barriers beyond the immediate control of PCP must be surmounted for the full integration of primary prevention in primary care practice. However, independent of other barriers, physician cognitive value of primary prevention in practice, a base mediator of physician behavior, will need to be increased to frame the likelihood of such integration. PMID- 12729466 TI - Weekly versus daily changes of in-line suction catheters: impact on rates of ventilator-associated pneumonia and associated costs. AB - BACKGROUND: An earlier randomized, controlled trial showed that weekly or as needed (as opposed to daily) changes of in-line suction catheters were associated with substantial cost savings, without a higher rate of ventilator-associated pneumonia (VAP). To examine the impact of decreasing the frequency of in-line suction catheter changes in our medical intensive care unit, we conducted an observational study, comparing the catheter costs and frequency of VAP during (1) a control period, during which in-line suction catheters were changed daily, and (2) a treatment period, during which the catheters were changed every 7 days or sooner if needed, for mechanical failure or soilage. METHODS: All adult patients admitted to our 18-bed medical intensive care unit were evaluated for the 3-month interval 1 year prior to the practice change (May through July 1998) and for the 3 months after implementing the new policy (May through July 1999). To avoid bias related to usual seasonal variation in VAP frequency, we also determined (via medical records) the VAP rate during May through July 1997. The occurrence of VAP was ascertained by an infection control practitioner, using criteria established by the Centers for Disease Control and applied in a standard fashion. The VAP rate was calculated as the mean number of VAPs per 100 ventilator-days for each 3 month interval. Use of ventilators, humidifiers, and non-heated-wire, disposable circuits was uniform during the study, as were policies regarding humidity, temperature settings, and frequency of routine ventilator circuit changes. RESULTS: During the control period 146 patients accounted for 1,075 ventilator days and there were 2 VAPs (0.19 VAPs per 100 ventilator-days). During the treatment period 143 patients accounted for 1,167 ventilator-days and there were no VAPs. The mean +/- SD duration of in-line suction catheter use during the treatment period was 3.8 +/- 0.8 days, and 51% of the patients had the same catheter in place for > 3 days (range 4-9 days). The actual cost of catheters used during the treatment period was lower than during the control period ($1,330 vs $6,026), predicting annual catheter cost savings of $18,782. CONCLUSIONS: We conclude that (1) a policy of weekly (vs daily) change of in-line suction catheter is associated with substantial cost savings, with no significant increase in the frequency of VAP, and (2) to the extent that these findings confirm the results of prior studies they support a policy of changing in-line suction catheters weekly rather than daily. PMID- 12729467 TI - Critical thinking in respiratory care practice: a qualitative research study. AB - INTRODUCTION: Recent publications indicate that critical thinking should be an integral part of respiratory care education. However, we know very little about critical thinking in the context of respiratory care. The critical thinking abilities and decision-making characteristics of practicing respiratory therapists have not been studied. PURPOSE: Identify and describe the critical thinking skills and traits of respiratory therapists, using a qualitative, descriptive research methodology. METHODS: Critical thinking was defined as the combination of logical reasoning, problem-solving, and reflection. The sample was selected through nominations of experts, using reputational-case selection. The research involved observations of 18 registered respiratory therapists, followed by in-depth interviews. Data were collected over a 1-year period and there were 125 hours of observation and 36 hours of interview. The observations were the basis for identifying and describing context-bound situations that require critical thinking, as well as the essential skills and related traits. RESULTS: The data set consists of over 600 single-spaced pages of interview transcripts and participant-observation field notes, in addition to 36 audio tapes. Field notes and interview transcripts were continuously analyzed throughout the study, using the constant-comparative method described by Glaser and Strauss. CONCLUSIONS: The findings suggest that critical thinking in respiratory care practice involves the abilities to prioritize, anticipate, troubleshoot, communicate, negotiate, reflect, and make decisions. PMID- 12729465 TI - Wnt/Wingless signaling through beta-catenin requires the function of both LRP/Arrow and frizzled classes of receptors. AB - BACKGROUND: Wnt/Wingless (Wg) signals are transduced by seven-transmembrane Frizzleds (Fzs) and the single-transmembrane LDL-receptor-related proteins 5 or 6 (LRP5/6) or Arrow. The aminotermini of LRP and Fz were reported to associate only in the presence of Wnt, implying that Wnt ligands form a trimeric complex with two different receptors. However, it was recently reported that LRPs activate the Wnt/beta-catenin pathway by binding to Axin in a Dishevelled--independent manner, while Fzs transduce Wnt signals through Dishevelled to stabilize beta-catenin. Thus, it is possible that Wnt proteins form separate complexes with Fzs and LRPs, transducing Wnt signals separately, but converging downstream in the Wnt/beta catenin pathway. The question then arises whether both receptors are absolutely required to transduce Wnt signals. RESULTS: We have established a sensitive luciferase reporter assay in Drosophila S2 cells to determine the level of Wg- stimulated signaling. We demonstrate here that Wg can synergize with DFz2 and function cooperatively with LRP to activate the beta-catenin/Armadillo signaling pathway. Double-strand RNA interference that disrupts the synthesis of either receptor type dramatically impairs Wg signaling activity. Importantly, the pronounced synergistic effect of adding Wg and DFz2 is dependent on Arrow and Dishevelled. The synergy requires the cysteine-rich extracellular domain of DFz2, but not its carboxyterminus. Finally, mammalian LRP6 and its activated forms, which lack most of the extracellular domain of the protein, can activate the Wg signaling pathway and cooperate with Wg and DFz2 in S2 cells. We also show that the aminoterminus of LRP/Arr is required for the synergy between Wg and DFz2. CONCLUSION: Our study indicates that Wg signal transduction in S2 cells depends on the function of both LRPs and DFz2, and the results are consistent with the proposal that Wnt/Wg signals through the aminoterminal domains of its dual receptors, activating target genes through Dishevelled. PMID- 12729468 TI - Radiographically occult right main bronchus intubation with a fastrach laryngeal mask airway endotracheal tube. AB - We report a case in which the distal 3 cm of an LMA-Fastrach laryngeal mask airway (LMA) endotracheal tube (ETT) was radiographically invisible. After the LMA intubation, left lung atelectasis developed. The radiopaque wire coil built into the ETT was mistakenly believed to mark the end of the ETT, so the radiograph made it appear that the end of the ETT was 2 cm above the main carina. In fact, this type of ETT extends 3 cm beyond the end of the wire coil, and the final 3 cm of the ETT can be difficult or impossible to see on a radiograph. Bronchoscopy revealed that the end of the ETT was in the bronchus intermedius. In this case, even in retrospect, the true end of the ETT could not be seen on the radiograph. Clinicians should be aware that the final 3 cm of the LMA-Fastrach ETT can be radiographically invisible. We believe the manufacturer should redesign this ETT to include better radiopaque markers all the way to the end of the ETT. PMID- 12729469 TI - Ventilator auto-triggering in a patient with tuberculous bronchopleural fistula. AB - We report a case of ventilator auto-triggering resulting from tuberculous bronchopleural fistula being managed with chest tube suction. Early recognition of bronchopleural fistula-related auto-triggering is extremely important. Auto triggering can lead to serious adverse effects, including severe hyperventilation and inappropriate escalation of sedatives and/or neuromuscular blockers (administered to reduce spontaneous breathing efforts). Auto-triggering was confirmed in our patient when tachypnea persisted despite pharmacologic neuromuscular paralysis. Auto-triggering can be reduced or eliminated by decreasing ventilator trigger sensitivity or by decreasing the air leak flow by reducing the degree of chest tube suction. PMID- 12729470 TI - An unusual complication of endotracheal intubation. PMID- 12729471 TI - A major role for non-major histocompatibility complex genes but not for microorganisms in a novel murine model of Graves' hyperthyroidism. AB - The etiology of Graves' disease is multifactorial. We investigated the role of genetic and environmental factors on the susceptibility to Graves' hyperthyroidism using a new murine model. Intramuscular injection of recombinant adenovirus expressing the thyrotropin receptor (AdCMVTSHR) induces Graves'-like hyperthyroidism (thyrotropin receptor [TSHR] antibodies, elevated thyroxine, and diffuse goiter) in more than 50% of female BALB/c mice. The relative contributions of major histocompatibility complex (MHC) and non-MHC genes on the susceptibility to hyperthyroidism were studied by immunizing BALB/c (H-2d), BALB.K (H-2k), and DBA/2J (H-2d) mice with AdCMVTSHR. Hyperthyroidism developed in approximately 50% of BALB/c and BALB.K mice but only 5% of DBA/2J mice, indicating a major role for non-MHC genes in disease development. The effect of environmental microorganisms was evaluated by comparing disease incidence in BALB/c mice maintained in pathogen-free conditions versus those in nonsterile, conventional housing, as well as by coadministering microorganism components (Escherichia coli lipopolysaccharide or yeast zymosan A) as adjuvants with AdCMVTSHR. Neither type of exposure to environmental pathogens influenced disease induction. In conclusion, non-MHC genes, but not infectious organisms, play a major role in the etiology of this novel murine model of Graves' disease. PMID- 12729472 TI - Activin betaB expression in rat experimental goiter and human thyroid tumors. AB - Activins are dimeric proteins of the transforming growth factor beta superfamily, which exhibit multiple functions in gonadal and extragonadal tissues. Expression of activin A, composed of two betaA subunits, has been shown in the thyroid, whereas there has been no study regarding activin B (betaBbetaB) in this gland. In other tissues, such as the gonads, pancreas, and adrenal cortex, expression of both activin betaA and activin betaB has been described. In this study, we detected activin betaB mRNA and protein expression using reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry in rat experimental goiter and in human thyroid, including multinodular goiter, follicular adenoma, papillary carcinoma, and follicular carcinoma. Activin betaA mRNA and protein expression was also investigated in rat and human thyroid tissue. The expression of both activin betaB and activin betaA was highest in rat methimazole-induced goiter and in human follicular adenoma, and papillary and follicular carcinomas when compared with multinodular goiter and normal thyroid tissue. The increased expression of activin betaB as well as activin betaA, observed in this study, suggests that activin B and activin A may be involved in the proliferative and neoplastic processes of the thyroid. PMID- 12729473 TI - Medroxyprogesterone acetate decreases secretion of interleukin-6 and parathyroid hormone-related protein in a new anaplastic thyroid cancer cell line, KTC-2. AB - A new thyroid cancer cell line, KTC-2, was established from the malignant pleural effusion of a patient with recurrent thyroid cancer associated with anaplastic transformation from thyroid papillary cancer. Karyotype analysis showed a mode of 109 chromosomes. Subcutaneous cell injections produced small regressing tumors in athymic or severe combined immunodeficiency disorders (SCID) mice. Histologic examination showed anaplastic tumor cells surrounded by prominent mononuclear cells. An expression of thyroglobulin, thyroid transcription factor-1, and PAX-8 but not thyroid peroxidase and thyrotropin (TSH) receptor was detected. Biochemical analysis revealed secretion of interleukin (IL)-6, parathyroid hormone-related protein (PTHrP), and granulocyte-macrophage colony-stimulating factor. All the cytokines are known to induce paraneoplastic syndromes in patients with anaplastic thyroid cancer. Our previous studies revealed that medroxyprogesterone acetate (MPA) reduces secretion of IL-6 and PTHrP from human breast cancer cells. To investigate the regulatory mechanisms of secretion of these cytokines, MPA was administered to the KTC-2 cells. MPA dose-dependently decreased the secretion and mRNA expression of IL-6 and PTHrP. Expression of androgen receptor and glucocorticoid receptor (GR) but not progesterone receptor was detected. Dexamethasone but not dihydrotestosterone and progesterone decreased IL-6 and PTHrP secretion. These findings suggest that MPA decreases IL 6 and PTHrP secretion as a glucocorticoid mediated by GR in the KTC-2 cells. This KTC-2 cell line may be a suitable model for developing new strategies against paraneoplastic syndromes caused by anaplastic thyroid cancer. PMID- 12729475 TI - Radioactive iodine and the salivary glands. AB - Radioactive iodine ((131)I) targets the thyroid gland and has been proven to play an effective role in the treatment of differentiated papillary and follicular cancers. Simultaneously, this radioisotope hones in on the salivary glands where it is concentrated and secreted into the saliva. Dose related damage to the salivary parenchyma results from the (131)I irradiation. Salivary gland swelling and pain, usually involving the parotid, can be seen. The symptoms may develop immediately after a therapeutic dose of (131)I and/or months later and progress in intensity with time. In conjunction with the radiation sialadenitis, secondary complications reported include xerostomia, taste alterations, infection, increases in caries, facial nerve involvement, stomatitis, candidiasis, and neoplasia. Prevention of the (131)I sialadenitis may involve the use of sialogogic agents to hasten the transit time of the radioactive iodine through the salivary glands. However, studies are not available to delineate the efficacy of this approach. Recently, amifostine has been advocated to prevent the effects of irradiation. Treatment of the varied complications that may develop encompass numerous approaches and include gland massage, sialogogic agents, duct probing, antibiotics, mouthwashes, good oral hygiene, and adequate hydration. PMID- 12729474 TI - Relation of CD30 molecules on T-cell subsets to the severity of autoimmune thyroid disease. AB - The prognosis of patients with autoimmune thyroid disease (AITD) varies. To clarify the immunologic differences among patients with various severities of AITD, we examined two types of molecules on peripheral T lymphocytes: CD195 (CCR5), which express dominantly on CD4(+) type 1 helper T (T(H)1) cells, and CD30, which is known as a marker of CD4(+) type 2 helper T (T(H)2) cells and a regulatory molecule of CD8(+) autoreactive cytotoxic T cells. We found presence of patients with high proportion (> 9%) of CD30 expression in CD4(+) cells in a group of patients with Graves' disease (GD) in remission compared to the patients with intractable GD and a decrease in the intensity of CD30 expression on CD8(+) cells from patients with severe Hashimoto's disease (HD) treated for hypothyroidism compared to patients with untreated and euthyroid HD. There was no difference in CD195 expression between these patients with GD or HD with different severities, but there was a decreased intensity of CD195(+) cells in thyrotoxic patients with GD. These results indicate that CD30 molecules on CD4(+) and CD8(+) cells may be related to the severities of GD and HD, respectively. PMID- 12729476 TI - Treatment of recurrent nodular goiters with percutaneous ethanol injection: a clinical study of twelve patients. AB - Twelve patients who had previously undergone thyroid surgery received percutaneous ethanol injection (PEI) treatment because of recurrent nodular goiter (3 with a toxic [TN], 2 with a nontoxic cystic [NCN], and 7 with a nontoxic solid nodule [NSN]). Two of the 12 had recurrent nerve palsy contralateral to the nodule. Each patient received a mean total dose of 0.88 mL of ethanol per milliliter of nodular volume. Ethanol was injected in a mean of 3.5 sessions for solid and 3 sessions for NCN. In most cases, a slight to moderate burning pain was experienced during and for 12-48 hours after PEI treatment, and one patient experienced temporary hoarseness. One patient with TN and 2 patients with NSN became hypothyroid, 7 patients with nontoxic nodules remained euthyroid, 1 with TN became euthyroid, and a previously hyperthyroid patient with TN became subclinically hyperthyroid 1-year posttherapy. The nodule shrank by more than 50% of the pretreatment volume in all patients (8.6 +/- 2.6 vs. 2.9 +/- 1.2 mL in TN, and 12.3 +/- 4.9 vs. 4.16 +/- 2.54 mL in nontoxic nodules, pretreatment vs. 1 year posttreatment volume, respectively). With regard to the increased risk of reoperation, PEI treatment can be proposed for patients with recurrent nodular goiter requiring surgery. PMID- 12729477 TI - Levothyroxine a new drug? Since when? How could that be? PMID- 12729478 TI - Anaplastic thyroid cancer: cytogenetic patterns by comparative genomic hybridization. AB - We studied chromosomal abnormalities by comparative genomic hybridization (CGH) and flow cytometry in anaplastic thyroid cancer (ATC), and when present in coexisting or previous differentiated thyroid cancer (DTC). Overall 10 frozen tissues from patients with ATC and 5 cell lines (1 ATC and 4 DTCs) were analyzed. We found chromosomal abnormalities in 5 of 10 ATC tissues, with 24 abnormalities (22 gains and 2 losses). Among 8 ATCs that were associated with prior or concurrent DTC, more chromosomal abnormalities were found in ATC associated with follicular thyroid cancer (FTC) than those associated with PTC (median numbers 9.5 and 0.5, respectively, p = 0.046) or no associated differentiated thyroid cancer. Gain of 1q was relatively common in ATCs (30%). By flow cytometry, we found aneuploidy in 6 of 10 ATC tissues and diploidy in 4. There was concordance between DNA aneuploidy and the presence of chromosomal abnormalities by CGH in 4 of the 5 ATCs (p = 0.048). We also found 26 chromosomal abnormalities in an ATC cell line, 14.3 in 3 FTC cell line, and 3 in a PTC cell line. In conclusion, chromosomal abnormalities are frequent in ATCs associated with FTC, but uncommon in those associated with PTC and in ATCs with no associated differentiated thyroid cancer. These findings support the concept that PTC and FTC have different genetic backgrounds and, even after the transformation to ATC, they may retain some of their cytogenetic characteristics. PMID- 12729479 TI - CD10 expression is useful in the diagnosis of follicular carcinoma and follicular variant of papillary thyroid carcinoma. AB - CD10/neutral endopeptidase 24.11(NEP) is a membrane-bound zinc metalloproteinase the expression of which represents a useful tool in the classification and diagnosis of malignant leukemia and lymphoma. Recently, CD10 has been found to be expressed in nonhematopoietic tissues and various types of neoplasms. In this study, we examined CD10 immunostaining in paraffin sections of 70 distinct lesions to investigate whether CD10 is a useful diagnostic marker for thyroid neoplasms. CD10 was not detected in normal thyroid tissue, benign lesions (15 follicular lesions and 15 adenomatous goiters), and pure papillary carcinomas except for follicular variants. In contrast, CD10 was expressed in 8 of 10 (80%) follicular carcinomas and 7 of 9 (77%) follicular variant of papillary thyroid carcinomas. This appears to be the first report on the expression of this member of a newly identified gene family in thyroid tumors. In conclusion, immunohistochemistry of CD10 in paraffin sections is valuable in the classification of thyroid follicular lesions into benign and malignant groups and in the diagnosis of follicular variant of papillary thyroid carcinoma. PMID- 12729480 TI - Familial aggregation of autoimmune thyroiditis in first-degree relatives of patients with juvenile autoimmune thyroid disease. AB - Several studies have shown aggregation of autoimmune thyroiditis in families by estimation of thyroid antibodies. However, the prevalence by concurrent estimation with fine-needle aspiration cytology (FNAC) and thyroid antibodies has not been previously reported. We therefore studied 222 first-degree relatives (group 1) of 71 index cases diagnosed as lymphocytic thyroiditis on FNAC and 81 family members (group 2) of 23 goitrous children diagnosed as colloid goiter on FNAC for comparison. Successful FNAC conducted in 122 group 1 subjects revealed lymphocytic thyroiditis in 51 (42%), whereas lymphocytic thyroiditis was diagnosed in only 5 goitrous subjects (13%) in group 2. Among group 1 subjects with FNAC-proven lymphocytic thyroiditis, antithyroid peroxidase (TPO) antibodies were found in 35 (67%), while in anti-TPO antibody positive goitrous relatives of group 1, lymphocytic thyroiditis was found in 36 (78%). Eight new cases of overt hypothyroidism and 45 new cases of subclinical hypothyroidism were diagnosed among group 1 subjects. Our study suggests: (1). familial clustering of autoimmune thyroiditis; (2). if only FNAC or thyroid antibodies is used for diagnosis of autoimmune thyroiditis in children, 22%-33% of cases are likely to be missed; and (3). serum thyrotropin (TSH) should be offered to all first-degree relatives of patients with juvenile autoimmune thyroiditis. PMID- 12729481 TI - Interleukin-6 and thyroid hormone metabolism in pediatric cardiac surgery patients. AB - Pediatric patients undergoing cardiac surgery have been reported to have low serum triiodothyronine (T(3)) levels in the postoperative period. The cause of this dysfunction is not known, although proinflammatory cytokines such as interleukin-6 (IL-6) have been implicated in the inhibition of hepatic conversion of thyroxine (T(4)) to T(3). This study measured serum levels of IL-6 and T(3) during the first 4 postoperative days in 16 children (mean age, 28 +/- 7 days) undergoing cardiopulmonary bypass surgery. The mean preoperative serum total T(3) level was 164 +/- 30 ng/dL (2.5 +/- 0.5 nmol/L) that decreased significantly to a nadir of 43 +/- 8 ng/dL (0.6 +/- 0.01 nmol/L) within 48 hours after surgery. Serum IL-6 levels increased significantly from 16 +/- 7 pg/mL preoperatively to a peak value of 374 +/- 134 pg/mL measured 2-3 hours after surgery. A positive correlation (r(2) = 0.507) was found between the peak serum level of IL-6 and the lowest serum T(3) level in each patient attained during the 4 postoperative days. Potential treatments directed toward diminishing the rise in proinflammatory cytokines in the immediate postoperative period may prove effective in preventing the low serum T(3) in children undergoing cardiac surgery, and thus diminish the associated postoperative morbidity. PMID- 12729482 TI - Ectopic intrathyroidal thymoma: a case report and review. AB - Ectopic intrathyroidal thymomas are an exceedingly rare clinical entity that can be challenging to diagnose. This report describes a 39-year-old Japanese woman who presented with prominent left-sided thyroid enlargement that was thought to be a dominant thyroid nodule by ultrasound. Two fine-needle aspiration biopsies showed an atypical lymphoid proliferation that was suspicious for although not diagnostic of a low-grade lymphoma. A diagnosis of ectopic intrathyroidal thymoma was made only after appropriate histopathologic assessment of the surgical specimen. PMID- 12729484 TI - Amyloidosis: role of fine-needle aspiration. PMID- 12729483 TI - Sclerosing lymphocytic lobulitis of the breast in a patient with Graves' disease. AB - A 43-year-old woman presented to the endocrinologist with symptoms and signs of typical thyrotoxicosis caused by Graves' disease. Review of systems revealed that she had recently discovered a lump in her left breast. Evaluation of the left breast lesion led to a core biopsy that showed sclerosing lymphocytic lobulitis. This breast disease, well recognized in the pathology literature, occurs in various autoimmine disorders, particularly type 1 diabetes mellitus, and has occasionally been reported in Hashimoto's thyroiditis. The patient described here represents the first published association of sclerosing lymphocytic lobulitis of the breast with Graves' disease. PMID- 12729485 TI - Parietal lobe contribution to mental rotation demonstrated with rTMS. AB - A large number of imaging studies have identified a role for the posterior parietal lobe, in particular Brodmann's area 7 and the intraparietal sulcus (IPS), in mental rotation. Here we investigated whether neural activity in the posterior parietal lobe is essential for successful mental rotation performance by observing the effects of interrupting this activity during the execution of a mental rotation task. Repetitive transcranial magnetic stimulation (rTMS) was applied to posterior parietal locations estimated to overlie Brodmann's area 7 in the right and the left hemisphere, or to a posterior midline location (sham condition). In three separate experiments, rTMS (four pulses, 20 Hz) was delivered at these locations either 200-400, 400-600, or 600-800 msec after the onset of a mental rotation trial. Disrupting neural activity in the right parietal lobe interfered with task performance, but only when rTMS was delivered 400 to 600 msec after stimulus onset. Stimulation of the left parietal lobe did not reliably affect mental rotation performance at any of the time points investigated. The time-limited effect of rTMS was replicated in a fourth experiment that directly compared the effects of rTMS applied to the right parietal lobe either 200-400 or 400-600 msec into the mental rotation trial. The results indicate that the right superior posterior parietal lobe plays an essential role in mental rotation, consistent with its involvement in a variety of visuospatial and visuomotor transformations. PMID- 12729486 TI - Characterization of empathy deficits following prefrontal brain damage: the role of the right ventromedial prefrontal cortex. AB - Impaired empathic response has been described in patients following brain injury, suggesting that empathy may be a fundamental aspect of the social behavior disturbed by brain damage. However, the neuroanatomical basis of impaired empathy has not been studied in detail. The empathic response of patients with localized lesions in the prefrontal cortex (n = 25) was compared to responses of patients with posterior (n = 17) and healthy control subjects (n = 19). To examine the cognitive processes that underlie the empathic ability, the relationships between empathy scores and the performance on tasks that assess processes of cognitive flexibility, affect recognition, and theory of mind (TOM) were also examined. Patients with prefrontal lesions, particularly when their damage included the ventromedial prefrontal cortex, were significantly impaired in empathy as compared to patients with posterior lesions and healthy controls. However, among patients with posterior lesions, those with damage to the right hemisphere were impaired, whereas those with left posterior lesions displayed empathy levels similar to healthy controls. Seven of nine patients with the most profound empathy deficit had a right ventromedial lesion. A differential pattern regarding the relationships between empathy and cognitive performance was also found: Whereas among patients with dorsolateral prefrontal damage empathy was related to cognitive flexibility but not to TOM and affect recognition, empathy scores in patients with ventromedial lesions were related to TOM but not to cognitive flexibility. Our findings suggest that prefrontal structures play an important part in a network mediating the empathic response and specifically that the right ventromedial cortex has a unique role in integrating cognition and affect to produce the empathic response. PMID- 12729487 TI - The effect of cingulate cortex lesions on task switching and working memory. AB - Anatomic interconnections between the prefrontal and anterior cingulate cortices suggest that these areas may have similar functions. Here we report the effect of anterior cingulate removal on task switching, error monitoring, and working memory. Neuroimaging studies have implicated the cingulate cortex in all these processes. Six macaques were taught task switching (TS) and delayed alternation (DA) paradigms. TS required switching between two conditional response tasks with mutually incompatible response selection rules. DA required alternation between two identically covered food-well positions. In the first set of experiments, anterior cingulate lesions did not consistently impair TS or DA performance. One animal performed worst on both TS and DA and in this animal the cingulate sulcus lesion was most complete. In the second set of experiments, we confirmed that larger anterior cingulate lesions, which included the sulcus, consistently impaired TS but only led to a mild and equivocal impairment of DA. The TS error pattern, however, did not suggest an impairment of TS per se. The consequence of a cingulate lesion is, therefore, distinct to that of a prefrontal lesion. TS error distribution analyses provided some support for a cingulate role in monitoring responses for errors and subsequent correction but the pattern of reaction time change in TS was also indicative of a failure to sustain attention to the task and the responses being made. PMID- 12729488 TI - A magnetic stimulation examination of orthographic neighborhood effects in visual word recognition. AB - The split-fovea theory proposes that visual word recognition is mediated by the splitting of the foveal image, with letters to the left of fixation projected to the right hemisphere (RH) and letters to the right of fixation projected to the left hemisphere (LH). We applied repetitive transcranial magnetic stimulation (rTMS) over the left and right occipital cortex during a lexical decision task to investigate the extent to which word recognition processes could be accounted for according to the split-fovea theory. Unilateral rTMS significantly impaired lexical decision latencies to centrally presented words, supporting the suggestion that foveal representation of words is split between the cerebral hemispheres rather than bilateral. Behaviorally, we showed that words that have many orthographic neighbors sharing the same initial letters ("lead neighbors") facilitated lexical decision more than words with few lead neighbors. This effect did not apply to end neighbors (orthographic neighbors sharing the same final letters). Crucially, rTMS over the RH impaired lead-, but not end-neighborhood facilitation. The results support the split-fovea theory, where the RH has primacy in representing lead neighbors of a written word. PMID- 12729489 TI - The right hemisphere involvement in the processing of morphologically derived words. AB - It is widely documented that the left hemisphere is dominant in all complex linguistic tasks, including the processing of inflectional morphology. Both in Italian and in other languages, patients with brain damage with a selective deficit in derivational morphology have never been reported. Here we present the unusual case of two patients with very similar right-hemisphere lesions, who in the absence of aphasic disorders showed a selective inability in producing derivational morphology. Although both patients were unimpaired in producing verb infinitives, they both showed a selective deficit in producing nouns derived from verbs. This difficulty was not present in deriving nouns from other grammatical categories, such as adjectives. Interestingly, both patients mostly substituted the derived noun with the past participle of the verb. This pattern of results documents for the first time a right-hemisphere contribution in the domain of derivational morphology. PMID- 12729490 TI - Neural correlates of lexical access during visual word recognition. AB - People can discriminate real words from nonwords even when the latter are orthographically and phonologically word-like, presumably because words activate specific lexical and/or semantic information. We investigated the neural correlates of this identification process using event-related functional magnetic resonance imaging (fMRI). Participants performed a visual lexical decision task under conditions that encouraged specific word identification: Nonwords were matched to words on orthographic and phonologic characteristics, and accuracy was emphasized over speed. To identify neural responses associated with activation of nonsemantic lexical information, processing of words and nonwords with many lexical neighbors was contrasted with processing of items with no neighbors. The fMRI data showed robust differences in activation by words and word-like nonwords, with stronger word activation occurring in a distributed, left hemisphere network previously associated with semantic processing, and stronger nonword activation occurring in a posterior inferior frontal area previously associated with grapheme-to-phoneme mapping. Contrary to lexicon-based models of word recognition, there were no brain areas in which activation increased with neighborhood size. For words, activation in the left prefrontal, angular gyrus, and ventrolateral temporal areas was stronger for items without neighbors, probably because accurate responses to these items were more dependent on activation of semantic information. The results show neural correlates of access to specific word information. The absence of facilitatory lexical neighborhood effects on activation in these brain regions argues for an interpretation in terms of semantic access. Because subjects performed the same task throughout, the results are unlikely to be due to task-specific attentional, strategic, or expectancy effects. PMID- 12729492 TI - Success and failure suppressing reflexive behavior. AB - The dynamic interplay between reflexive and controlled determinants of behavior is one of the most general organizing principles of brain function. A powerful analogue of this interplay is seen in the antisaccade task, which pits reflexive and willed saccadic mechanisms against one another. Event-related functional magnetic resonance imaging of the human brain showed greater prestimulus preparatory activity in the pre-supplementary motor area before voluntary antisaccades (saccades away from a target) compared with reflexive prosaccades (saccades to a target). Moreover, this preparatory activity was critically associated with reflex suppression; it predicted whether the reflex was later successfully inhibited in the trial. These dataillustrate a mechanism for top down control over reflexive behavior. PMID- 12729491 TI - A parametric manipulation of factors affecting task-induced deactivation in functional neuroimaging. AB - Task-induced deactivation (TID) refers to a regional decrease in blood flow during an active task relative to a "resting" or "passive" baseline. We tested the hypothesis that TID results from a reallocation of processing resources by parametrically manipulating task difficulty within three factors: target discriminability, stimulus presentation rate, and short-term memory load. Subjects performed an auditory target detection task during functional magnetic resonance imaging (fMRI), responding to a single target tone or, in the short term memory load conditions, to target sequences. Seven task conditions (a common version and two additional levels for each of the three factors) were each alternated with "rest" in a block design. Analysis of covariance identified brain regions in which TID occurred. Analyses of variance identified seven regions (left anterior cingulate/superior frontal gyrus, left middle frontal gyrus, right anterior cingulate gyrus, left and right posterior cingulate gyrus, left posterior parieto-occipital cortex, and right precuneus) in which TID magnitude varied across task levels within a factor. Follow-up tests indicated that for each of the three factors, TID magnitude increased with task difficulty. These results suggest that TID represents reallocation of processing resources from areas in which TID occurs to areas involved in task performance. Short-term memory load and stimulus rate also predict suppression of spontaneous thought, and many of the brain areas showing TID have been linked with semantic processing, supporting claims that TID may be due in part to suspension of spontaneous semantic processes that occur during "rest" (Binder et al., 1999). The concept that the typical "resting state" is actually a condition characterized by rich cognitive activity has important implications for the design and analysis of neuroimaging studies. PMID- 12729493 TI - Face-selective activation in a congenital prosopagnosic subject. AB - Congenital prosopagnosia is a severe impairment in face identification manifested from early childhood in the absence of any evident brain lesion. In this study, we used fMRI to compare the brain activity elicited by faces in a congenital prosopagnosic subject (YT) relative to a control group of 12 subjects in an attempt to shed more light on the nature of the brain mechanisms subserving face identification. The face-related activation pattern of YT in the ventral occipito temporal cortex was similar to that observed in the control group on several parameters: anatomical location, activation profiles, and hemispheric laterality. In addition, using a modified vase-face illusion, we found that YT's brain activity in the face-related regions manifested global grouping processes. However, subtle differences in the degree of selectivity between objects and faces were observed in the lateral occipital cortex. These data suggest that face related activation in the ventral occipito-temporal cortex, although necessary, might not be sufficient by itself for normal face identification. PMID- 12729494 TI - The X-trials: neural correlates of an inhibitory control task in children and adults. AB - Event-related potentials (ERPs) were used to examine developmental differences between adults and 6-year-old children in the neural processes involved in an inhibitory control task. Twenty adults and 21 children completed a task that required them to selectively respond to target stimuli while inhibiting responses to equally salient non-target stimuli. Because this task had been previously studied using functional magnetic resonance imaging (fMRI), the relation between the fMRI and ERP findings was informally examined. The results indicate that latency and amplitude of the P3 differentiated the different types of trials. However, the pattern of event-related neural activity differed for adults and children. These results, which suggest that adults and children may be using different processes to perform this task, have implications for the interpretation of the previous fMRI findings. PMID- 12729495 TI - Adult brain plasticity elicited by anomia treatment. AB - We describe a study where a specific treatment method for word-finding difficulty (so-called contextual priming technique, which combines massive repetition priming with semantic priming) was applied with three chronic left hemisphere damaged aphasics. Both before and after treatment, which focused on naming of a series of pictures, naming-related brain activity was measured by magnetoencephalography (MEG). Due to its excellent temporal resolution and good spatial resolution, we were able to track treatment-induced changes in cortical activity. All three subjects showed improved naming of the trained items. In all subjects, a single source area, located in the left inferior parietal lobe, close to the lesioned area, displayed statistically significant training-induced changes. This effect was of long latency as it started 300-600 msec after picture presentation. The change in activation was specific to training, as it could not be accounted for by variation of cortical dynamics associated with increased proportion of correct answers. Our interpretation is that the training effect reflects more effective phonological encoding and storage of the trained items through the engagement of a left hemispheric word-learning system. This is in line with recent functional imaging studies, which have linked left inferior parietal lobe activity to the phonological storage component of the verbal working memory, as well as with theoretical arguments stating that the primary role of the phonological loop is to acquire new words. Finally, the MEG results showed no evidence of increased right hemisphere participation following training, supporting the view that restoration of language-related networks in the damaged left hemisphere is crucial for anomia recovery. PMID- 12729496 TI - Neural correlates of the left-visual-field superiority in face perception appear at multiple stages of face processing. AB - Studies in healthy individuals and split-brain patients have shown that the representation of facial information from the left visual field (LVF) is better than the representation of facial information from the right visual field (RVF). To investigate the neurophysiological basis of this LVF superiority in face perception, we recorded event-related potentials (ERPs) to centrally presented face stimuli in which relevant facial information is present bilaterally (B faces) or only in the left (L faces) or the right (R faces) visual field. Behavioral findings showed best performance for B faces and, in line with the LVF superiority, better performance for L than R faces. Evoked potentials to B, L, and R faces at 100- to 150-msec poststimulus showed no evidence of asymmetric transfer of information between the hemispheres at early stages of visual processing, suggesting that this factor is not responsible for the LVF superiority. Neural correlates of the LVF superiority, however, were manifested in a shorter latency of the face-specific N170 component to L than R faces and in a larger amplitude to L than R faces at 220-280 and 400-600 msec over both hemispheres. These ERP amplitude differences between L and R faces covaried across subjects with the extent to which the face-specific N170 component was larger over the right than the left hemisphere. We conclude that the two hemispheres exchange information symmetrically at early stages of face processing and together generate a shared facial representation, which is better when facial information is directly presented to the right hemisphere (RH; L faces) than to the left hemisphere (LH; R faces) and best when both hemispheres receive facial information (B faces). PMID- 12729497 TI - Lexical and sublexical components of age-related changes in neural activation during visual word identification. AB - Positron emission tomography data (Madden, Langley, et al., 2002) were analyzed to investigate adult age differences in the relation between neural activation and the lexical (word frequency) and sublexical (word length) components of visual word identification. The differential influence of these components on reaction time (RT) for word/nonword discrimination (lexical decision) was generally similar for the two age groups, with word frequency accounting for a greater proportion of lexical decision RT variance relative to word length. The influence of word length on RT, however, was relatively greater for older adults. Activation in regions of the ventral occipito-temporal cortex was related to the RT changes associated with word frequency and length for older adults, but not for younger adults. Specifically, older adults' frequency effects were related to activation in both anterior (Brodmann's area [BA] 37) and posterior (BAs 17 and 18) regions of the occipito-temporal pathway, whereas word length effects were only associated with posterior activation (BA 17). We conclude that aging affects the neural mechanisms supporting word identification performance although behavioral measures of this ability are generally constant as a function of age. PMID- 12729498 TI - Determinants of the annual pattern of reproduction in mature male Merino and Suffolk sheep: responses to a nutritional stimulus in the breeding and non breeding seasons. AB - This study was designed to test whether an acute improvement in diet would increase gonadotrophin secretion and testicular growth in strongly photoperiod responsive Suffolk rams and weakly photoperiod-responsive Merino rams in both the breeding (February-March) and the non-breeding (July-August) seasons. Mature rams (n = 5 or 6) of these breeds were fed a maintenance diet (0.9 kg chaff + 100 g lupin grain) or the same diet supplemented with 1.5 kg lupin grain for 42 days in each season. Lupin grain is a rich source of both energy and protein. Testosterone, luteinizing hormone (LH) and follicle stimulating hormone (FSH) were measured in plasma from blood sampled every 20 min for 24 h on Days -1, 12 and 35 relative to the change in feeding. In rams supplemented with lupins, body mass increased in both breeds in both seasons (P < 0.001). Scrotal circumference and LH pulse frequency increased with lupin supplementation in both seasons (P < 0.003) in Merinos, but only during the breeding season (P < 0.003) in Suffolks. Plasma FSH concentrations were affected by diet only during the breeding season, being elevated on Day 12 in lupin-supplemented rams of both breeds (P < 0.05). It was concluded that Merino rams exhibit reproductive responses to improved nutrition irrespective of time of the year, whereas Suffolk rams respond to nutrition only when the hypothalamic reproductive centres are not inhibited by photoperiod. Thus, Suffolks do respond to nutrition, just as Merinos do, but only when photoperiod allows. This difference between breeds appears to be a result of differences in the neuroendocrine pathways that control pulsatile gonadotrophin releasing hormone secretion. PMID- 12729500 TI - Superoxide dismutase affects the viability of thawed European mouflon (Ovis g. musimon) semen and the heterologous fertilization using both IVF and intracytoplasmatic sperm injection. AB - This study evaluated the effects of superoxide dismutase (SOD) on viability and acrosome integrity of European mouflon spermatozoa after cryopreservation and on the fertilization rates of sheep oocytes after i.v.f. or intracytoplasmatic sperm injection (i.c.s.i.). Frozen semen was thawed and washed with synthetic oviduct fluid supplemented with 0.6% bovine serum albumin. After centrifugation, the spermatozoa pellet was split into two culture systems: (i) without SOD; and (ii) in the presence of 1500 IU mL(-1) SOD. Sperm viability and acrosome integrity were evaluated simultaneously, immediately after thawing and after 3, 6 and 9 h of culture (5% CO2, 39 degrees C, 90% humidity), by incubating sperm with propidium iodide and fluorescein isothiocyanate-labelled Pisum sativum agglutinin. At the same time, sperm were assessed for motility using a standard scoring system (independent operators' observation of sperm) that graded degree of motility (i.e. 1 = immotile to 10 = maximum motility, as observed at the moment of thawing). For i.v.f., frozen-thawed semen derived from the two culture systems was placed in culture together with in vitro-matured sheep oocytes. For i.c.s.i., semen derived from the same culture systems as that for i.v.f. was used, and incubated for 1 h under standard conditions. The results showed a marked difference (P < 0.01) between the percentages of live spermatozoa in medium with SOD and those obtained in medium alone, after 3, 6 and 9 h of culture. The percentages of intact acrosome spermatozoa were higher in medium with SOD after 6 h (P = 0.05) of culture. Spermatozoa motility decreased significantly in SOD containing medium at 3 and 6 h of culture compared with motility in control medium. Fertilization rates were significantly lower in medium with SOD than in medium alone, whereas in the i.c.s.i. system fertilization rates were significantly higher in the presence of SOD. The results indicate that the addition of SOD to the culture media enhances the viability rates and the acrosome integrity of cryopreserved mouflon spermatozoa. PMID- 12729499 TI - Effect of dietary intake on steroid feedback on release of luteinizing hormone in ovariectomized cows. AB - This study tested the hypothesis that the decline in pulsatile release of luteinizing hormone (LH), resulting from steroid negative feedback, is greater in animals fed a low, compared with a high, plane of nutrition. Two-year-old cows were ovariectomized and six days later were fed diets to provide 1.5 x maintenance requirements (n = 6, supplemented) or 0.5 x maintenance requirements (n = 6, restricted) (Round 1). Pulsatile release of LH was measured over a 14-h period on the fifth day of feeding these diets (Day 1); at 6 h, all animals were treated with an intravaginal insert containing 1.38 g progesterone, which remained in place until the end of Day 3. Pulsatile release of LH was again measured for 14 h on Day 3; at 6 h, all animals were injected intramuscularly with oestradiol benzoate (ODB; 1 mg per 500 kg live weight). Three days later, this protocol was repeated, in a cross-over design, with cows that were previously restricted now being supplemented and those cows previously supplemented, now restricted (Round 2). Plasma concentrations of progesterone after intravaginal progesterone treatment were 1.01 ng mL(-1) higher in restricted cows compared with supplemented cows (P < 0.001) and were also higher in Round 1 than in Round 2 and on Day 1 than on Day 3 (P < 0.001). Plasma concentrations of oestradiol following injection with ODB did not differ between supplemented and restricted cows (P > 0.1). Dietary intake did not affect mean concentrations of LH, pulse frequency or amplitude during the 6-h period before steroid treatment or the change in these variables following steroid treatment; however, the slope of the decline in concentrations of LH following progesterone treatment was significantly more negative in cows fed restricted diets compared with those fed supplemented diets. In Round 2, mean concentrations of LH were higher preceding, and decreased more following, progesterone treatment compared with the decrease after ODB treatment. In conclusion, acute dietary restriction resulted in a more rapid decline in the release of LH following treatment with intravaginal progesterone, and was associated with higher concentrations of progesterone in plasma. PMID- 12729502 TI - The role of insulin-like growth factor II and its receptor in mouse preimplantation development. AB - Insulin-like growth factor II (IGF-II) and its receptor, the IGF-II/mannose-6 phosphate (IGF-II/M6P) receptor, are first expressed from the zygotic genome at the two-cell stage of mouse development. However, their role is not clearly defined. Insulin-like growth factor II is believed to mediate growth through the heterologous type 1 IGF and insulin receptors, whereas the IGF-II/M6P receptor is believed to act as a negative regulator of somatic growth by limiting the availability of excess levels of IGF-II. These studies demonstrate that IGF-II does have a role in growth regulation in the early embryo through the IGF-II/M6P receptor. Insulin-like growth factor II stimulated cleavage rate in two-cell embryos in vitro. Moreover, this receptor is required for the glycaemic response of two-cell embryos to IGF-II and for normal progression of early embryos to the blastocyst stage. Improved development of embryos in crowded culture supports the concept of an endogenous embryonic paracrine activity that enhances cell proliferation. These responses indicate that the IGF-II/M6P receptor is functional and likely to participate in such a regulatory circuit. The functional role of IGF-II and its receptor is discussed with reference to regulation of early development. PMID- 12729501 TI - Prolactin signalling in porcine theca cells: the involvement of protein kinases and phosphatases. AB - The hypothesis that protein kinase C (PKC) and tyrosine kinases, as well as serine-threonine and tyrosine phosphatases, are involved in prolactin (PRL) signalling in theca cells harvested from porcine follicles was tested. Theca cells were incubated with PRL for 24 h to stimulate progesterone (P4) production. In addition, treatments included inhibitors of PKC and tyrosine kinases, as well as serine-threonine phosphatase inhibitor and tyrosine phosphatase inhibitor. Prolactin significantly stimulated P4 production by theca cells and all inhibitors suppressed the PRL-stimulated P4 production. After incubation with PRL for 2, 5, 10 or 20 min, theca cells were homogenized and cytosolic and membrane fractions were obtained. This was followed by determination of PKC activity in partially purified subcellular fractions by measuring the transfer of 32P from [gamma-32P] adenosine triphosphatase (ATP) to histone III-S. In unstimulated porcine theca cells the major proportion of PKC activity was present in the cytosol. Incubation of cells with PRL resulted in a rapid, time-dependent increase in the amount of PKC activity in the membrane fraction. Protein kinase C activity in the membrane fraction was maximal after 10 min of cells' exposure to PRL. Protein kinase C activation was assessed also by measuring the specific association of 3H-phorbol dibutyrate (3H-PDBu) with theca cells after treatment with PRL. Prolactin significantly increased 3H-PDBu-specific binding in theca cells. In contrast to PKC, total inositol phosphate accumulation was not affected by PRL in the current study. In summary, PRL stimulated P4 production by porcine theca cells derived from large follicles. The results of the study were consistent with the hypothesis that PKC is one of the intracellular mediators of PRL action in porcine theca cells. Protein kinase C activation does not appear to occur through the action of phosphatidylinositol-dependent phospholipase C. Moreover, the involvement of tyrosine kinases, as well as tyrosine and serine threonine phosphatases, in PRL signalling in the examined cells is suggested. PMID- 12729503 TI - Reproduction and plasma concentrations of leptin, insulin and insulin-like growth factor 1 in growth-hormone-transgenic female sheep before and after artificial insemination. AB - The transgenic sheep used in this study expressed an additional copy of the gene for ovine growth hormone (GH), so they had continuously high plasma concentrations of GH. They were used to test whether the GH transgene affected plasma concentrations of the metabolic hormones leptin, insulin-like growth factor 1 (IGF-1) and insulin, and whether these effects were associated with changes in conception, pregnancy or parturition following artificial insemination. Compared with control animals, the GH-transgenic sheep had higher bodyweight, lower body condition score and less subcutaneous fat (P < 0.05). These sheep also had lower plasma concentrations of leptin, higher plasma concentrations of insulin, and higher plasma concentrations of IGF-1 (P < 0.001). A similar proportion of GH-transgenic and control ewes came into oestrus, but the conception rate to artificial insemination was lower in GH-transgenic ewes than in the controls. Only four live lambs were recovered from 12 GH-transgenic ewes (33%) compared with 38 lambs from 43 controls (88%). This outcome was not associated with any difference in plasma progesterone profile in the period leading up to artificial insemination (Day 0). The GH-transgenic ewes had lower concentrations of FSH at all times measured (Day -19, Day -2 and Day 19). These results indicate that appropriate regulation of GH secretion from pituitary or peripheral tissues is necessary for normal reproduction and normal levels of metabolic hormones. Chronically high concentrations of GH were associated with increased levels of IGF-1 and insulin, and decreased levels of leptin. PMID- 12729504 TI - The pattern of surfactant cholesterol during vertebrate evolution and development: does ontogeny recapitulate phylogeny? AB - Pulmonary surfactant is a complex mixture of phospholipids (PLs), neutral lipids and proteins that lines the inner surface of the lung. Here it modulates surface tension, thereby increasing lung compliance and preventing the transudation of fluid. In humans, pulmonary surfactant is comprised of approximately 80% PLs, 12% neutral lipids and 8% protein. In most eutherian (i.e. placental) mammals, cholesterol (Chol) comprises approximately 8-10% by weight or 14-20 mol% of both alveolar and lamellar body surfactant. It is regarded as an integral component of pulmonary surfactant, yet few studies have concentrated on its function or control. The lipid composition is highly conserved within the vertebrates, except that surfactant of teleost fish is dominated by cholesterol, whereas tetrapod pulmonary surfactant contains a high proportion of disaturated phospholipids (DSPs). The primitive Australian dipnoan lungfish Neoceratodus forsterii demonstrates a 'fish-type' surfactant profile, whereas the other derived dipnoans demonstrate a surfactant profile similar to that of tetrapods. Homology of the surfactant proteins within the vertebrates points to a single evolutionary origin for the system and indicates that fish surfactant is a 'protosurfactant'. Among the terrestrial tetrapods, the relative proportions of DSPs and cholesterol vary in response to lung structure, habitat and body temperature (Tb), but not in relation to phylogeny. The cholesterol content of surfactant is elevated in species with simple saccular lungs or in aquatic species or in species with low Tb. The DSP content is highest in complex lungs, particularly of aquatic species or species with high Tb. Cholesterol is controlled separately from the PL component in surfactant. For example, in heterothermic mammals (i.e. mammals that vary their body temperature), the relative amount of cholesterol increases in cold animals. The rapid changes in the Chol to PL ratio in response to various physiological stimuli suggest that these two components have different turnover rates and may be packaged and processed differently. In mammals, the pulmonary surfactant system develops towards the end of gestation and is characterized by an increase in the saturation of PLs in lung washings and the appearance of surfactant proteins in amniotic fluid. In general, the pattern of surfactant development is highly conserved among the amniotes. This conservation of process is demonstrated by an increase in the amount and saturation of the surfactant PLs in the final stages (>75%) of development. Although the ratios of surfactant components (Chol, PL and DSP) are remarkably similar at the time of hatching/birth, the relative timing of the maturation of the lipid profiles differs dramatically between species. The uniformity of composition between species, despite differences in lung morphology, birthing strategy and relationship to each other, implies that the ratios are critical for the onset of pulmonary ventilation. The differences in the timing, on the other hand, appear to relate primarily to birthing strategy and the onset of air breathing. As the amount of cholesterol relative to the phospholipids is highly elevated in immature lungs, the pattern of cholesterol during development and evolution represents an example of ontogeny recapitulating phylogeny. The fact that cholesterol is an important component of respiratory structures that are primitive, when they are not in use or developing in an embryo, demonstrates that this substance has important and exciting roles in surfactant. These roles still remain to be explored. PMID- 12729505 TI - Lipid metabolism in the embryos of diabetic rats during early organogenesis: modulatory effect of prostaglandin E2. AB - The purpose of this work was to evaluate de novo lipid biosynthesis and the lipid profile, and to study the effect of prostaglandin E2 (PGE2; prostaglandin has previously been found to be involved in diabetes embryopathy) on lipid metabolism in embryos from control and streptozotocin-induced diabetic rats during organogenesis. Increased levels of triacylglycerols were found in embryos of diabetic rats compared with controls, whereas no differences were detected in the levels of cholesterol, cholesterylester, phosphatidylcholine and phosphatidylethanolamine. When the de novo synthesis of lipids in the embryo was studied using [14C]acetate as a tracer, a diminished rate of incorporation of [14C]acetate into the evaluated lipid classes was detected in the diabetic embryo compared with controls. Addition of PGE2 did not modify the incorporation of [14C]acetate into any of the lipid species of control embryos, but enhanced the incorporation of [14C]acetate into triacylglycerol, cholesterylesters, phosphatidylcholine and phosphatidylethanolamine of embryos from diabetic rats. The study's results show alterations in both synthesis and concentrations of lipids in the embryos of diabetic rats. Interestingly, the results demonstrate that the addition of PGE2, a prostaglandin that reverses the embryonic morphological abnormalities induced by diabetes, prevents disturbances in embryo lipid synthesis caused by diabetes. PMID- 12729506 TI - A population-based comparison between travelers who consulted travel clinics and those who did not. AB - BACKGROUND: Travel to hepatitis A-endemic countries is frequent among North Americans. Such travel carries significant risks for the individuals themselves and for the general population. We documented the patterns of use of travel clinics in a large Canadian adult population. METHODS: Travelers who had visited a hepatitis A-endemic country between 1990 and the time of the survey in 1999 were eligible. Subjects were identified from a representative sample of 4,002 adults from the two largest Canadian provinces. They were contacted by random digit dialing and interviewed by telephone. RESULTS: Only 15% of trips had been preceded by a visit to a travel clinic. The probability of visiting a travel clinic was approximately 10 times greater for travelers considered to be in the high-risk category than for those in the low-risk category, but the former represented only 2% of the total. The probability of visiting a travel clinic was approximately 23 times greater for travelers who were aware of the health risks in their country of destination. Income level was not associated with attendance at a travel clinic, and cost was rarely mentioned as a reason for not attending such a travel clinic before departure. CONCLUSIONS: Each year, millions of Canadian travelers go to hepatitis A-endemic countries without consulting a travel clinic. Active steps must be taken by public health authorities to improve their utilization of health services and prevent the accrued health risk for these travelers. PMID- 12729509 TI - Cosmic radiation and commercial air travel. PMID- 12729507 TI - Immunity to hepatitis A in people born and raised in endemic areas. AB - BACKGROUND: Hepatitis A is the most common vaccine-preventable disease in international travelers. Many individuals born and raised in hepatitis A endemic areas are likely to be immune to hepatitis A. Unnecessary hepatitis A immunization could be avoided by taking into account prior exposure to hepatitis A and judicious use of serotesting prior to immunization. METHODS: Patients born and raised in countries of high prevalence of hepatitis A who were seen for pretravel consultation and who had hepatitis antibody measured were eligible. Data were collected about country of birth and length of residence there before emigration, length of time till departure on current trip, current age, and hepatitis A antibody result. RESULTS: Patients ranged from 12 to 74 years of age and were from 27 countries. Their pretravel visit occurred from 4 to more than 90 days prior to departure. Ninety-five percent (122 of 129) of patients were immune to hepatitis A, including 100% (83 of 83) of those who resided in their country of origin until at least aged 20. Most patients were seen for pretravel consultation less than 28 days prior to departure. CONCLUSION: Individuals born and raised until aged 20 in hepatitis A endemic countries are likely to be immune to hepatitis A. Serotesting is most helpful in assessing immunity to hepatitis A in those under 20 years of age. PMID- 12729508 TI - Resources utilized by general practitioners for advising travelers from Australia. PMID- 12729510 TI - Prevention and treatment of toxic seafoodborne diseases in travelers. PMID- 12729512 TI - New slaves and malaria. PMID- 12729511 TI - Travelers' diarrhea in the new millennium: consensus among experts from German speaking countries. PMID- 12729513 TI - When should travelers from nonendemic areas for flaviviruses receive booster vaccination for Japanese encephalitis? PMID- 12729514 TI - Should contacts of patients with rabies be advised to seek postexposure prophylaxis? A survey of tropical medicine experts. PMID- 12729515 TI - Leptospirosis in travelers returning from the Dominican Republic. PMID- 12729516 TI - W-135 meningococcal disease in a traveler: a case report. PMID- 12729517 TI - Tick paralysis by Ixodes holocyclus in a Japanese traveler returning from Australia associated with Rickettsia helvetica infection. PMID- 12729518 TI - Chronic malaria with splenic rupture. PMID- 12729519 TI - Epidemiologic rigor in travel medicine. PMID- 12729521 TI - Travel medicine in Turkey: current state and implications for the future. PMID- 12729522 TI - [Prognostic factors for mortality in out-of-hospital cardiorespiratory arrest]. AB - BACKGROUND AND OBJECTIVE: Our purpose was to determine the prognostic factors of mortality among patients with cardiorespiratory arrest (CRA) assisted by 061 emergency teams in Andalusia. PATIENTS AND METHOD: Retrospective observational study from January 1998 to December 1999 of 1950 cases of out-of-hospital CRA (OH CRA) assisted by Andalusian 061 emergency teams (ETs). Independent (predictor) variables considered in the study were those defined in the Utstein style, after categorization. The dependent (outcome) variable was out-of-hospital mortality. A multivariate model was constructed using logistic regression to define the factors that, when considered together, predict mortality. The model was calibrated using the Hosmer-Lemeshow test. For the discrimination of the model, we calculated the area under the ROC curve. RESULTS: The incidence of OH-CRA was 27/100,000. Among our population of 1950 patients, 24.95% (483) were admitted alive to hospital and 75.05% (1444) died in the out-of hospital setting; 1393 patients were male and 552 were female. The mean age was 61.3 28.4 years. The model revealed the following mortality prognostic factors: personnel performing cadiopulmonary resuscitation before ET arrival (healthcare/non-healthcare), presence of defibrillation, number of defibrillations, CRA site, general function categories before CRA, and cardiac massage within the first minute by ET. CONCLUSIONS: In order to reduce the CRA-induced mortality in our setting, defibrillation and cardiac massage by ETs must be done without delay. It is fundamental to achieve greater health awareness and education among both the general population and the healthcare workers involved in the survival chain. PMID- 12729523 TI - [Isolated dietary counselling program versus supplement and dietary counselling in patients with human immunodeficiency virus infection]. AB - BACKGROUND AND OBJECTIVE: Nutritional intervention is a controversial area. The aim of the study was to compare the influence on nutritional status of nutritional supplementation with a standard polymeric formula as well as nutritional counselling versus isolated nutritional counselling in a group of HIV infected patients. PATIENTS AND METHOD: There were 70 patients, 66 of whom were fully evaluated for each study end point after application of prospectively determined evaluability criteria. Of these, 35 were randomized to group I (standard formula) and 35 were randomized to unsupplemented group II. Group I patients received standard enteral formula (3 cans/day, 250 ml per can). Patients were submitted to a prospective serial assessment of their nutrition status (anthropometric and biochemical status) and the nutritional intake was determined by means of 24-hours written food records. Determinations were performed at baseline and at 3 months. RESULTS: Treatment with oral supplements in group I resulted in a significant and sustained increase in weight (2.75%; p < 0.05) which was mostly due to fat mass (10.8%; p < 0.05). In contrast, no changes were detected in group II patients. The increase in body weight and tricipital skinfold was significant in group I. Total body water and fat free-mass remained without changes. CD4 counts and viral load remained stable in both groups. CONCLUSIONS: Oral nutritional supplements for a 3-months period resulted in body weight gain in HIV-infected patients, increasing the fat mass. An isolated nutritional counselling did not result in such an increase. PMID- 12729524 TI - [Population-based reference values for the Spanish version of the SF-36 Health Survey in the elderly]. AB - BACKGROUND AND OBJECTIVE: The SF-36 Health Survey questionnaire is the most widely used instrument to measure health-related quality of life. Reference measures are needed to interpret its results in clinical and population studies. In 1996, a study provided population-based norms for Spain, but these were not disaggregated by age in subjects aged 75 years and over, even though health status changes with aging among those who overpass such age. Moreover, health status in elderly people from developed countries has improved over the last years. This study obtains population-based norms for the Spanish version of the SF-36 Health Survey in five-year age-groups for those aged 60 to 85 and over. SUBJECTS AND METHOD: Cross-sectional survey on a sample of 3,949 non institutionalised subjects representative of the Spanish population aged 60 years and over. Information on the SF-36 Health Survey was obtained through house-hold personal interviews. Central position and dispersion statistics, as well as percentiles, were calculated for each of the eight SF-36 scales by age and sex. Cronbach's alpha coefficients were calculated to assess the internal reliability of each scale. RESULTS: Subjects reported higher scores for emotional role (mean [SD] 84.8 [32.9]), social functioning (79.2 [28.0]) and physical role (73.3 [41.1]). For all scales, mean scores were higher (better perceived health) among men than women (p < 0.0001). As age increased, mean scores in all scales decreased (p for linear trend < 0.001) except for mental health (p for linear trend = 0.29 in women, p for lineal trend = 0.14 in men), yet the decrease was greater for physical functioning and physical role. Reliability was very high for all scales (Cronbach's alpha from 0.84 to 0.95). RESULTS were similar to those of the study carried out in 1996. CONCLUSIONS: These results extend those obtained in 1996 and facilitate the interpretation of the SF-36 Health Survey values in clinical and population studies in the Spanish population aged 60 years and older. PMID- 12729526 TI - [Active prevention for pressure ulcers]. PMID- 12729525 TI - [Drug bradyarrhythmias as a cause of hospital admission. Study of 83 cases]. AB - BACKGROUND AND OBJECTIVE: The frequency and circumstances associated with drugs induced bradiarrhythmia (DB) causing hospital admission are little known. PATIENTS AND METHOD: Retrospective analyses of DB in Cardiology (CD) and Internal Medicine (IMD) Departments of a 3rd level hospital over two consecutive years. RESULTS: We detected 83 cases (mean age: 72.68, 75.9% women). Most frequent DB was slow atrial fibrillation (55.4%). DB were due to the use of one drug in 41 cases and to an association in 42 cases. Most frequently involved drug was digoxine (62.7%). DB were due to the use of one drug in 41 cases and to an association in 42 cases. Most frequently involved drug was digoxine (62.7%), which was commonly associated with another drug (69.3%). Most frequent pharmacological associations were negative chronotropic drugs (57.1%) followed by their association with a diuretic and/or an angiotensin converting enzyme inhibitor (42.8%), causing renal failure and/or potassium imbalance. CONCLUSIONS: Drugs-induced bradiarrhythmia is a frequent cause of hospital admission in aged people, especially women, and is frequently due to drugs associations and/or renal failure. PMID- 12729527 TI - [Hypersensitivity pneumonitis in construction plasterers (espartosis): study of 20 patients]. AB - BACKGROUND AND OBJECTIVE: Espartosis is a type of hypersensitivity pneumonitis which frequently affects construction workers handling esparto fibres used as support material inside gypsum plaster. Exactly which agents produce this disease remains under discussion. The aims of this study were: a) to assess the possible etiologic role of the fungi colonizing esparto grass fibres and esparto itself in the genesis of this disease, and b) to describe the clinical characteristics of this disease in a large series of patients. PATIENTS AND METHOD: Twenty patients diagnosed of hypersensitivity pneumonitis due to esparto grass exposure were studied. Mycologic cultures of the esparto grass fibre samples provided by each patient were performed. Fungi and/or esparto were used for determination of specific IgG antibodies, specific skin tests and specific bronchial challenge tests. RESULTS: Most frequently isolated fungi in causal esparto samples were Aspergillus sp. (60%) and Mucor sp. (47%). Specific IgG antibody determinations and/or specific bronchial challenge tests showed antigenicity not only for Aspergillus sp. but also for non-fungi-contaminated esparto grass and other fungi such as Penicillium sp. or Mucor sp. These results were helpful for establishing a new etiologic approach to the diagnosis of this disease. CONCLUSIONS: Aspergillus fumigatus is recognized as a causal agent in hypersensitivity pneumonitis due to esparto grass exposure; however, other antigenic sources such as Penicillium frequentans and other fungi, as well as esparto grass fibres, also appear to play a role in the genesis of this disease. PMID- 12729528 TI - [Adult cystic fibrosis: new clinical presentations]. PMID- 12729529 TI - [Pain at the right groin and cranial osteolysis in a 74-year-old woman]. PMID- 12729531 TI - [Severe hyponatremia and intranasal desmopressin-associated water intoxication in a female patient with diabetes insipidus and histiocytosis X]. PMID- 12729530 TI - [Malignant pleomorphic fibrous histocytoma of the pericardium]. PMID- 12729532 TI - [Renal failure as a complication of acute Q fever]. PMID- 12729533 TI - [Skin tuberculosis due to direct inoculation]. PMID- 12729534 TI - [Oil spills and public health]. PMID- 12729537 TI - [Impact of smoking and alcohol consumption on mortality in the population of Barcelona [Spain]: 1983-1998]. AB - OBJECTIVES: To analyze the mortality attributable to smoking and alcohol consumption in the city of Barcelona from 1983 to 1998. METHODS: All deaths among residents of the city of Barcelona from 1983 to 1998 were included. Population data were obtained from the city inhabitants register. The mortality attributable to smoking and alcohol consumption was calculated by population attributable fractions using relative risks from previous studies in the US population. RESULTS: In 1998, 2,205 deaths were attributable to smoking, representing 13.8% of all deaths among the population aged 35 years or older and a decrease of 9.6% compared with deaths in 1983. From 1983 to 1998 there was an increase in smoking attributable adjusted mortality rates for lung cancer (155.84/100,000 inhabitants in men and 9.39/100,000 in women in 1998) and chronic obstructive pulmonary disease (COPD) (95.89/100,000 in men and 11.29/ 100,000 in women in 1998). In 1998, deaths attributed to alcohol consumption accounted for 4.3% of total mortality, representing a reduction of 26% since 1983. Among men, the primary cause of alcohol-attributable mortality was liver cirrhosis (17.1%), although its relative importance decreased (accounting for 17.1% of alcohol-attributable mortality in 1998 compared with 24.3% in 1983). CONCLUSIONS: The mortality attributable to smoking and alcohol consumption decreased in the city of Barcelona during the study period. Smoking-attributable mortality from lung cancer and COPD increased in both sexes. Alcohol consumption-attributable mortality from liver cirrhosis decreased in men. PMID- 12729536 TI - [Back pain in the general population of Catalonia (Spain). Prevalence, characteristics and therapeutic behavior]. AB - BACKGROUND: Back pain is a common symptom of health assistance in Western countries. The goal of the present survey was to establish the prevalence of back pain in the general population in Catalonia, as well as to establish its characteristics, its relationship to sociodemographic variables, the therapeutic behavior followed by those affected and the associated-impairment and disabilities for the sufferers. METHODS: The survey was a descriptive study including a sample of 1,964 people, obtained from the census (1991) and representative of the population older than 18 years. The study was done in 1994 and data were obtained by means of a telephone interview. The prevalence of pain in the last six months was determined. Among those suffering back pain, additional information was obtained concerning its characteristics, the relationship with socio-demographic variables, the therapeutic behaviors used by patients, and the personal, social and work impairment and disabilities. RESULTS: Back pain was highly prevalent (50.9%), appeared in all ages (mean age of 47.6 years), was the highest in women (60.7%), in manual workers (54.9%) and in those less educated (71.1%). Back pain was long lasting (69.2% more than 3 years), frequent (49.7% more than the half of the days), and highly painful (severe unbearable in 51.4%). The therapeutic behaviors most commonly used were the visit to the physician (71.9%), the use of alternative medical treatments (24.7%), physical therapy (22.7%, physical exercises and electrotherapy) and self medication (14.6%, being acetylsalicylic acid the most employed). In general, the duration and degree of improvement was variable. Back pain limited the daily activities (36.7%) and forced to bed rest (22.7%). It was also a significant reason for time off work (17%) and disability pension (6.5%). CONCLUSIONS: Back pain has a very high prevalence in the Catalonian population and is an important reason to seek medical attention. It greatly limits the daily personal and professional activities. PMID- 12729538 TI - [Effectiveness of medical advice for reducing excessive alcohol consumption. Meta analysis of Spanish studies in primary care]. AB - INTRODUCTION: Former systematic reviews have backed the efficacy of medical counselling, a form of brief intervention, on the treatment of excessive drinkers detected in primary care settings. Nevertheless, these results cannot be applied without criticism to Mediterranean populations which, so far, have not been represented in the aforementioned studies. The aim of the present study was to update the results on the efficacy of brief interventions in primary care by pooling Spanish studies. METHODS: Studies were searched for by using appropriate databases and also by consulting to experts in the field to retrieve grey literature. Pooled estimations of effect sizes were calculated for two outcomes, the reduction in the amount of alcohol consumption and the decrease in the number of excessive drinkers. RESULTS: Two over the 5 retrieved studies were not included in a former review. The effect size regarding the decrease of alcohol consumption was medium (d = 0.46; 95% CI, 0.29 to 0.63; p < 0.0005; the intervention group outperformed the control by a 22%) and small for the decrease in the frequency of excessive drinkers (OR = 1.55; 95% CI, 1.06 to 2.26; p = 0.02; the intervention group outperformed the control by a 11%). The analysis by complimented protocols at the end of the study showed an effect size 1.5 times larger than the analysis performed on intention-to-treat basis. CONCLUSIONS: The results of this meta-analysis support the efficacy of brief intervention for excessive drinkers in primary care settings in Spain. PMID- 12729539 TI - [Methadone maintenance programs with supplementary services: a cost-effectiveness study]. AB - OBJECTIVE: Methadone maintenance programs (MMP) currently offer the best treatment for opioid-addicted patients. The aim of this study was to examine the cost-effectiveness of three MMPs that offered varying levels of supplementary services. Health-related quality of life was used as a measure of effectiveness. METHODS: A 12-month follow-up study of 586 patients beginning methadone treatment in Drug Care Centers in Barcelona was performed. The Nottingham Health Profile was used to measure quality of life. Standard unit costs and total cost per patient were calculated from activity registries. Sociodemographic, health related and toxicological data were collected through a semi-structured interview. A cost-effectiveness analysis was performed through two multiple linear regressions with the same adjusting variables. RESULTS: The greater the number of supplementary services involved, the higher the costs. The adjusted models revealed a significant increase in health-related quality of life (an increase of 8% in the Nottingham Health Profile) and in costs (17%) between low- and medium-intensity programs. CONCLUSION: The medium-intensity program showed the best cost-effectiveness ratio. However, the study's limitations preclude categoric generalization of the data. PMID- 12729540 TI - [Cost analysis and potential savings related to the use of preoperative tests in the hospitals of the Canary Islands [Spain]]. AB - OBJECTIVE: To estimate the potential cost savings that might take place in the Canary Islands Health Service (CHS) through the appropriate scientific use of preoperative tests, using information obtained from five public university hospitals. METHODS: A questionnaire was completed by 55 anesthesiologists, representing 60% of the total in the CHS. The questionnaire paid special attention to the most frequently used preoperative tests: chest radiograph, electrocardiogram, laboratory tests, and spirometry. The mean unit costs for the various preoperative tests were obtained from the two hospitals participating in the study. To calculate the potential cost savings derived from the appropriate use of preoperative tests, several scenarios were considered. These were characterized by different hypotheses or degrees of fulfillment of a protocol based on scientific knowledge and considered as the gold standard. RESULTS: In the ideal scenario in which the recommended scientific protocol was fulfilled in 100% of the 16 179 patients with an American Society of Anesthesiologists (ASA) grade of I-II, the economic impact would be notable, since it would free sensitive resources that could be used for other health programs. These figures could amount to approximately 1.02 million euros, without considering the cost of preoperative hospital stay. This figure could increase by up to approximately 2.13 million euros if one day of preoperative hospital stay were included and by up to 3.24 million euros if two days of preoperative hospital stay were included. CONCLUSIONS: The recent literature review and the results of the questionnaire applied in the CHS indicate that preoperative tests are of greatest benefit to patients and to society if their use is guided by scientific knowledge. In addition, resources can be freed by better selection and utilization of preoperative tests. PMID- 12729541 TI - [Trihalomethanes in the drinking water of Carabobo State, Venezuela]. AB - OBJECTIVE: Disinfection of water with chlorine in water treatment plants leads to the formation of trihalomethanes (THMs). These compounds are associated with adverse health effects. The aim of this study was to analyze THM concentrations in the water provided for human consumption in the two main water treatment systems of Carabobo State: the Alejo Zuloaga plant and the Pao-Cachinche reservoir, which form the Central Regional System I (CRS I), and the Lucio Baldo Soules plant and the Pao-La Balsa reservoir, which form the Central Regional System II (CRS II). METHODS: We analyzed 144 water samples collected in 6 samplings carried out in 2000 and 2001. THM concentrations were determined by gas chromatography using the headspace technique. The concentrations of the following THMs were measured: chloroform (CHCl3), bromoform (CHBr3), chlorodibromomethane (CHBr2Cl) and bromodichloromethane (CHCl2Br). RESULTS: The concentration of total THMs was between 47.84 g/l and 94.23 g/l. CHCl3 was the most commonly formed compound representing 83% of all THMs in the CRS I and 82% in the CRS II. The concentrations of total THMs in the CRS I, specifically in the Baja and San Diego networks, were significantly higher (p < 0.05) than permissible levels set by the American Environmental Protection Agency (80 g/l) for the sum of all four THMs. CONCLUSIONS: The results show that in the area studied there is a risk of adverse health effects due to THMs in drinking water, especially in the Baja and San Diego networks. PMID- 12729543 TI - [A personal balance of 25 years of modern health care management in the Spanish Public Health System]. AB - At the beginning of 2002, the transference of authority over public health care services from the Spanish national health System (INSALUD) to all the autonomous communities hitherto lacking this authority closed a period of modern halthcare management in Spain which, nevertheless, had lacked the dynamism and innovative capacity of business management. Despite this, during these 25 years, Spanish healthcare management underwent a spectacular growth. The problems of the Spanish public health system are either linked to the inherent problems of a system of universal coverage, with generous provisions and clear underfinancing, or they are characteristic of an obsolete organizational model in which centers and healthcare professionals lack autonomy, or they are related to users' increased expectations and to the enormous complexity of the medicine of the future. All these questions should be approached from the perspective of a new form of management appropriate to the new century. Despite its serious defects and problems, the public values the Spanish public health system; the main risk is chronic and progressive delegitimization wich causes users to lose faith and trust in the public health system. PMID- 12729542 TI - [Generic drugs: good or bad? Physician's knowledge of generic drugs and prescribing habits]. AB - In this article we analyze the responses of 1220 Spanish physicians who participated in a survery about generic drugs. A previously validated questionnaire was sent to physicians through the Spanish Medical Councils of the different provinces. Four items were analyzed: what doctors know about generic drugs (knowledge); physicians' prescribing habits concerning these drugs (attitude and professional competence); how prescription of generic drugs effects pharmaceutical costs amd, finally, what doctors believe a generic drug should be. The influence of physician-related variables (age, type of contract, specialty, workload, etc.) on prescribing of generic drugs was also analyzed. In view of the results, we believe that to rationalize expenditure through and appropriate policy on generic drugs Spanish health authorities should offer more and better training and information (clear and independent) about what generic drugs are. PMID- 12729544 TI - [The globalization of health]. AB - In this article diverse aspects of the relationship between health and globalization are explored. Different dimensions of globalization (economic, technological, cultural and political) are considered. Aspects of its effects on health (epidemiological, ethical and environmental), as well as its relationship with public health, power distribution and equity are discussed. Data that demonstrate the globalization of risks and of diseases, due to the current model of international relations and geographical mobility, are analyzed. The article defends the globalization of health and integrates renewed concepts and scientific advances in public health with politics, social strategies and new organizational forms of the practice of public health. Finally, we discuss the opportunities that have been provided by globalization since the middle of the last century for redefining world government and for developing local movements, based on solidarity and a new concept of politics, which could favor the universalization of health. PMID- 12729545 TI - [Tularemia possibly transmitted by red swamp crayfish]. PMID- 12729546 TI - [Control of Chagas' disease in Guarani communities: knowledge and hygiene habits within the Project to Improve Living Conditions in Bolivia]. AB - The aim of this study was to identify knowledge and control of vectorial transmission (Triatoma infestans, known as vinchuca) of Chagas' disease in Guarani Communities in Bolivia. We performed a descriptive study of a series of 98 individuals through a semi-structured questionnaire. Interviewees were asked about their familiarity with vinchuca, whether they thought vinchuca produced disease, the name of the disease and its consequences, as well as behavior related to eliminating the domestic insect vectors, such as cleaning of the home, backyard and corral.The insect vector was sufficiently well known (98%), although the name of the disease was identified by only 14.3% of the interviewees. Although the dwellings favored insect proliferation, they were not frequently cleaned: 28.6% cleaned their homes while and 42.9% cleaned the backyard and 7.1% cleaned the corral. Gender differences were found in the division of labor: women cleaned the homes and backyards, while men clean the corral. Experience has shown that the usefulness of projects for building healthy living areas and for health education depends on the value given to these projects by the community. Women are probably the best target group, because they perform a greater number of preventive tasks and seldom leave the community for extended periods of time. PMID- 12729547 TI - [Time to apply the precautionary principle]. PMID- 12729549 TI - A generalized estimating equations approach to quantitative trait locus detection of non-normal traits. AB - To date, most statistical developments in QTL detection methodology have been directed at continuous traits with an underlying normal distribution. This paper presents a method for QTL analysis of non-normal traits using a generalized linear mixed model approach. Development of this method has been motivated by a backcross experiment involving two inbred lines of mice that was conducted in order to locate a QTL for litter size. A Poisson regression form is used to model litter size, with allowances made for under- as well as over-dispersion, as suggested by the experimental data. In addition to fixed parity effects, random animal effects have also been included in the model. However, the method is not fully parametric as the model is specified only in terms of means, variances and covariances, and not as a full probability model. Consequently, a generalized estimating equations (GEE) approach is used to fit the model. For statistical inferences, permutation tests and bootstrap procedures are used. This method is illustrated with simulated as well as experimental mouse data. Overall, the method is found to be quite reliable, and with modification, can be used for QTL detection for a range of other non-normally distributed traits. PMID- 12729550 TI - Comparison of three multitrait methods for QTL detection. AB - A comparison of power and accuracy of estimation of position and QTL effects of three multitrait methods and one single trait method for QTL detection was carried out on simulated data, taking into account the mixture of full and half sib families. One multitrait method was based on a multivariate function as the penetrance function (MV). The two other multitrait methods were based on univariate analysis of linear combination(s) (LC) of the traits. One was obtained by a principal component analysis (PCA) performed on the phenotypic data. The second was based on a discriminate analysis (DA). It calculates a LC of the traits at each position, maximising the ratio between the genetic and the residual variabilities due to the putative QTL. Due to its number of parameters, MV was less powerful and accurate than the other methods. In general, DA better detected QTL, but it had lower accuracy for the QTL effect estimation when the detection power was low, due to higher bias than the other methods. In this case, PCA was better. Otherwise, PCA was slightly less powerful and accurate than DA. Compared to the single trait method, power can be improved by 30% to 100% with multitrait methods. PMID- 12729551 TI - Survival analysis of longevity in dairy cattle on a lactation basis. AB - An analysis of longevity in dairy cattle on a lactation basis is proposed. The approach allowed each lactation to have its own baseline hazard function, which gives a better description of the hazard than traditional analyses of the whole length of life. As a consequence, the overall fit of the model to the data was improved and fewer time-dependent variables were needed. Longevity on a lactation basis was defined from one calving to the next instead of from the first calving to culling. However, no new information was added and it was still the overall risk of being culled that was modelled. It is shown that no cow effect is needed in the lactation basis model because a censored record is not complete, a cow can appear as uncensored only once, and a cow cannot be censored after having been culled. Different subdivisions of the stage of lactation effect were tested and the first ten days of lactation were shown to correspond to an increased risk of being culled. There were no major differences in sire variance between the longevity analysed on a lactation basis and longevity based on the entire length of life. PMID- 12729552 TI - Combined analysis of data from two granddaughter designs: A simple strategy for QTL confirmation and increasing experimental power in dairy cattle. AB - A joint analysis of five paternal half-sib Holstein families that were part of two different granddaughter designs (ADR- or Inra-design) was carried out for five milk production traits and somatic cell score in order to conduct a QTL confirmation study and to increase the experimental power. Data were exchanged in a coded and standardised form. The combined data set (JOINT-design) consisted of on average 231 sires per grandsire. Genetic maps were calculated for 133 markers distributed over nine chromosomes. QTL analyses were performed separately for each design and each trait. The results revealed QTL for milk production on chromosome 14, for milk yield on chromosome 5, and for fat content on chromosome 19 in both the ADR- and the Inra-design (confirmed within this study). Some QTL could only be mapped in either the ADR- or in the Inra-design (not confirmed within this study). Additional QTL previously undetected in the single designs were mapped in the JOINT-design for fat yield (chromosome 19 and 26), protein yield (chromosome 26), protein content (chromosome 5), and somatic cell score (chromosome 2 and 19) with genomewide significance. This study demonstrated the potential benefits of a combined analysis of data from different granddaughter designs. PMID- 12729553 TI - Genetic structure of Balearic honeybee populations based on microsatellite polymorphism. AB - The genetic variation of honeybee colonies collected in 22 localities on the Balearic Islands (Spain) was analysed using eight polymorphic microsatellite loci. Previous studies have demonstrated that these colonies belong either to the African or west European evolutionary lineages. These populations display low variability estimated from both the number of alleles and heterozygosity values, as expected for the honeybee island populations. Although genetic differentiation within the islands is low, significant heterozygote deficiency is present, indicating a subpopulation genetic structure. According to the genetic differentiation test, the honeybee populations of the Balearic Islands cluster into two groups: Gimnesias (Mallorca and Menorca) and Pitiusas (Ibiza and Formentera), which agrees with the biogeography postulated for this archipelago. The phylogenetic analysis suggests an Iberian origin of the Balearic honeybees, thus confirming the postulated evolutionary scenario for Apis mellifera in the Mediterranean basin. The microsatellite data from Formentera, Ibiza and Menorca show that ancestral populations are threatened by queen importations, indicating that adequate conservation measures should be developed for protecting Balearic bees. PMID- 12729554 TI - Constructing the mammalian neocortex: the role of intrinsic factors. AB - The mammalian neocortex is subdivided into regions that are specialised for the processing of particular forms of information. These regions are distinct in terms of their cytoarchitecture, electrophysiology, and connectivity. How this regional diversity is generated through development is currently a topic of considerable interest and has centered upon two main issues. First, to what extent are these regions prespecified by intrinsic genetic mechanisms? Second, what is the influence of extrinsic activity in transmitting signals that ultimately shape functional regions? Historically, experimental evidence has tended to emphasise the role of extrinsic influences, but the identification and analysis of several genes that are expressed asymmetrically in the developing neocortex have tempered this viewpoint. We review current literature from the standpoint that intrinsic influences act early in neocortical development to generate molecular patterning whose main role is the guidance of long-range projections from the dorsal thalamus. Extrinsic influences appear to generate receptive fields for peripheral input, the summation of which determines the areal extent of particular neocortical region. PMID- 12729555 TI - Signals from the cerebellum guide the pathfinding of inferior olivary axons. AB - During development, inferior olivary axons cross the floor plate and project from the caudal to the rostral hindbrain, whence they grow into the cerebellar plate. We have investigated the axon guidance signals involved in the formation of this projection in vitro. When the cerebellar plate was grafted ectopically along the margin of the hindbrain in organotypic cultures, inferior olivary axons could pathfind to the ectopic cerebellum, establishing a topographically normal projection. Following rostrocaudal reversal of a region of tissue in the axon pathway between the inferior olive and the cerebellum, olivary axons still navigated towards the cerebellum. Moreover, olivary axons could cross a bridging tissue explant (spinal cord) to reach a cerebellar explant. In collagen gel cultures of inferior olive explants, olivary axon outgrowth increased significantly in the presence of cerebellar explants and axons deflected towards the cerebellar tissue. These results show that the cerebellum is a source of diffusible axon guidance signals for olivary axons. We also found that, in organotypic cultures, olivary axons which had crossed the floor plate showed an increased tendency to respond to cerebellar cues. Taken together, these results indicate that the cerebellum is the source of cues that are chemoattractant and growth-promoting for inferior olivary axons; prior exposure to the floor plate increases responsiveness to these cues. PMID- 12729556 TI - Posterior migration of the salivary gland requires an intact visceral mesoderm and integrin function. AB - The final overall shape of an organ and its position within the developing embryo arise as a consequence of both its intrinsic properties and its interactions with surrounding tissues. Here, we focus on the role of directed cell migration in shaping and positioning the Drosophila salivary gland. We demonstrate that the salivary gland turns and migrates along the visceral mesoderm to become properly oriented with respect to the overall embryo. We show that salivary gland posterior migration requires the activities of genes that position the visceral mesoderm precursors, such as heartless, thickveins, and tinman, but does not require a differentiated visceral mesoderm. We also demonstrate a role for integrin function in salivary gland migration. Although the mutations affecting salivary gland motility and directional migration cause defects in the final positioning of the salivary gland, most do not affect the length or diameter of the salivary gland tube. These findings suggest that salivary tube dimensions may be an intrinsic property of salivary gland cells. PMID- 12729557 TI - dHAND-Cre transgenic mice reveal specific potential functions of dHAND during craniofacial development. AB - Most of the bone, cartilage, and connective tissue of the craniofacial region arise from cephalic neural crest cells. Presumably, patterning differences in crest cells are a result of regional action of transcription factors within the developing pharyngeal arches. The basic helix-loop-helix transcription factor dHAND/HAND2 is expressed throughout much of the neural crest-derived mesenchyme of the pharyngeal arches, suggesting that it plays a crucial role in craniofacial development. However, targeted inactivation of the dHAND gene results in embryonic lethality by E10.5 due to vascular defects, preventing further analysis of the role of dHAND in cephalic neural crest cell development. In order to examine putative roles of dHAND during later stages of embryogenesis, we have used a transgenic lineage marker approach, in which a portion of the dHAND upstream region containing an enhancer that directs dHAND expression to the pharyngeal arches is used to drive Cre recombinase expression. By crossing these dHAND-Cre transgenic mice with R26R mice, we can follow the fate of cells that expressed dHAND at any time during development by examining beta-galactosidase activity. We show that dHAND is first expressed in postmigratory cephalic neural crest cells within the pharyngeal arches. In older embryos, beta-galactosidase labeled cells are observed in most of the neural crest-derived lower jaw skeleton and surrounding connective tissues. However, labeled cells only contribute to substructures within the middle ear, indicating that our transgene is not globally expressed in cephalic neural crest cells within the pharyngeal arches. Moreover, dHAND-Cre mice will provide a valuable tool for tissue-specific inactivation of gene expression in multiple tissue types of neural crest origin. PMID- 12729558 TI - Xenopus X-box binding protein 1, a leucine zipper transcription factor, is involved in the BMP signaling pathway. AB - We describe a novel basic leucine zipper transcription factor, XXBP-1, which interacts with BMP-4 in a positive feedback loop. It is a maternal factor and is zygotically expressed in the dorsal blastopore lip and ventral ectoderm with the exception of the prospective neural plate during gastrulation. Overexpression of XXBP-1 leads to ventralization of early embryos as described for BMP-4, and inhibits neuralization of dissociated ectoderm. Consistent with mediating BMP signaling, we show that the ectopic expression of XXBP-1 recovers the expression of epidermal keratin and reverses the dorsalization imposed by truncated BMP receptor type I, indicating that it may act downstream of the BMP receptor. Its effects can be partially mimicked by a fusion construct containing the VP16 activator domain and the XXBP-1 DNA-binding domain. In contrast, fusing the DNA binding domain to the even-skipped repressor domain leads to upregulation of the neural markers NCAM and nrp-1 in animal cap assay. Taken together, the results suggest a role for XXBP-1 in the control of neural differentiation, possibly as an activator. PMID- 12729559 TI - Analysis of the molecular cascade responsible for mesodermal limb chondrogenesis: Sox genes and BMP signaling. AB - Here, we have studied how Sox genes and BMP signaling are functionally coupled during limb chondrogenesis. Using the experimental model of TGFbeta1-induced interdigital digits, we dissect the sequence of morphological and molecular events during in vivo chondrogenesis. Our results show that Sox8 and Sox9 are the most precocious markers of limb cartilage, and their induction is independent and precedes the activation of BMP signaling. Sox10 appears also to cooperate with Sox9 and Sox8 in the establishment of the digit cartilages. In addition, we show that experimental induction of Sox gene expression in the interdigital mesoderm is accompanied by loss of the apoptotic response to exogenous BMPs. L-Sox5 and Sox6 are respectively induced coincident and after the expression of Bmpr1b in the prechondrogenic aggregate, and their activation correlates with the induction of Type II Collagen and Aggrecan genes in the differentiating cartilages. The expression of Bmpr1b precedes the appearance of morphological changes in the prechondrogenic aggregate and establishes a landmark from which the maintenance of the expression of all Sox genes and the progress of cartilage differentiation becomes dependent on BMPs. Moreover, we show that Ventroptin precedes Noggin in the modulation of BMP activity in the developing cartilages. In summary, our findings suggest that Sox8, Sox9, and Sox10 have a cooperative function conferring chondrogenic competence to limb mesoderm in response to BMP signals. In turn, BMPs in concert with Sox9, Sox6, and L-Sox5 would be responsible for the execution and maintenance of the cartilage differentiation program. PMID- 12729560 TI - Activation of Gbetagamma signaling downstream of Wnt-11/Xfz7 regulates Cdc42 activity during Xenopus gastrulation. AB - Wnt-11/Xfz7 signaling plays a major role in the regulation of convergent extension movements affecting the dorsal marginal zone (DMZ) of gastrulating Xenopus embryos. In order to provide data concerning the molecular targets of Wnt 11/Xfz7 signals, we have analyzed the regulation of the Rho GTPase Cdc42 by Wnt 11. In animal cap ectoderm, Cdc42 activity increases as a response to Wnt-11 expression. This increase is inhibited by pertussis toxin, or sequestration of free Gbetagamma subunits by exogenous Galphai2 or Galphat. Activation of Cdc42 is also produced by the expression of bovine Gbeta1 and Ggamma2. This process is abolished by a PKC inhibitor, while phorbol esther treatment of ectodermal explants activates Cdc42 in a PKC-dependent way, implicating PKC downstream of Gbetagamma. In activin-treated animal caps and in the embryo, interference with Gbetagamma signaling rescues morphogenetic movements inhibited by Wnt-11 hyperactivation, thus phenocopying the dominant negative version of Cdc42 (N(17)Cdc42). Conversely, expression of Gbeta1gamma2 blocks animal cap elongation. This effect is reversed by N(17)Cdc42. Together, our results strongly argue for a role of Gbetagamma signaling in the regulation of Cdc42 activity downstream of Wnt-11/Xfz7 in mesodermal cells undergoing convergent extension. This idea is further supported by the observation that expression of Galphat in the DMZ causes severe gastrulation defects. PMID- 12729561 TI - Inhibition of alpha4-integrin stimulates epicardial-mesenchymal transformation and alters migration and cell fate of epicardially derived mesenchyme. AB - Epithelial-mesenchymal transformation of the embryonic epicardium produces the subepicardial mesenchyme that is essential for normal coronary vascular development. Gene targeting experiments in mice have demonstrated an essential role for alpha4-integrin in normal epicardial development, but the precise cellular consequences of alpha4-integrin loss remain uncertain. To better understand the function of alpha4-integrin in epicardial development, we constructed a replication-incompetent adenovirus (AdlacZalpha4AS) that expresses antisense chicken alpha4-integrin as the 3' untranslated region of a lacZ reporter gene. This construct effectively labeled cells while greatly reducing levels of alpha4-integrin mRNA and protein. In quail chick chimeras, transplanted epicardial cells infected with AdlacZalpha4AS adhered to the heart and were incorporated into the epicardium, but 4 days after grafting, were largely absent from the epicardial epithelium, recapitulating the defect in alpha4-null mice. This did not result from epicardial cell apoptosis or anomalous migration of epicardial cells to extracardiac sites. Rather, AdlacZalpha4AS-infected epicardial cells were particularly invasive, being three to four times more likely to migrate to the interstitium of the myocardium than AdlacZ-infected epicardial cells. Accelerated epicardial-mesenchymal transformation and migration of alpha4-negative epicardium was observed in an organ culture system that does not require prior culture of epicardial cells. Remarkably, AdlacZalpha4AS infection also prevented targeting of epicardially derived mesenchyme to the media of developing coronary vasculature in the myocardial interstitium. This study provides evidence that epicardial alpha4-integrin normally restrains epicardial-mesenchymal transformation, invasion, and migration and is essential for correct targeting of epicardially derived mesenchyme to the developing coronary vasculature. PMID- 12729562 TI - The function of Xenopus germ cell nuclear factor (xGCNF) in morphogenetic movements during neurulation. AB - The germ cell nuclear factor (GCNF, NR6A1) is a nuclear orphan receptor first described in the mouse testis and subsequently identified as an essential transcription factor in vertebrate embryogenesis. Here, we analyze the phenotype of Xenopus embryos after depletion of embryonic GCNF (xEmGCNF) protein using a specific morpholino antisense oligonucleotide. Morphological defects after xEmGCNF knockdown became obvious from neurulation onward. Among the abnormalities observed, defective formation of the neural tube and a short and curved main body axis were the most remarkable traits. Histological analysis, lineage tracing of injected blastomeres, and Keller sandwich explants revealed that xEmGCNF function is required for different patterns of cell intercalation during neurulation and consequently for the sequence of morphogenetic movements leading to formation of the neural tube. Further characterization of the phenotype at the molecular level showed an abnormal distribution of the extracellular matrix protein fibronectin and a reduction in the expression level of the integrin subunits alpha5 and alpha6, the limiting components of the laminin and fibronectin receptors, respectively. We propose integrin-mediated cell-matrix interaction as a process that requires xEmGCNF function and provides, in concert with cadherins-mediated cell-cell interactions, a molecular basis for morphogenetic cell movements during neurulation. PMID- 12729564 TI - Regulation of metanephric kidney development by growth/differentiation factor 11. AB - Growth/differentiation factor 11 (Gdf11) is a transforming growth factor beta family member previously shown to control anterior/posterior patterning of the axial skeleton. We now report that Gdf11 also regulates kidney organogenesis. Mice carrying a targeted deletion of Gdf11 possess a spectrum of renal abnormalities with the majority of mutant animals lacking both kidneys. Histological analysis revealed a failure in ureteric bud formation at the initial stage of metanephric development in most Gdf11 mutant embryos examined. The metanephric mesenchyme of mutant embryos lacking a ureteric bud was found to be defective in the expression of glial cell line-derived neurotrophic factor (Gdnf), a gene known to direct ureteric bud outgrowth. The addition of Gdnf protein to urogenital tracts taken from Gdf11 null embryos induced ectopic ureteric bud formation along the Wolffian duct. Our studies suggest that Gdf11 may be important in directing the initial outgrowth of the ureteric bud from the Wolffian duct by controlling the expression of Gdnf in the metanephric mesenchyme. PMID- 12729563 TI - The amino-terminal region of Gli3 antagonizes the Shh response and acts in dorsoventral fate specification in the developing spinal cord. AB - A concentration gradient of Shh is thought to pattern the ventral neural tube, and these ventral cell types are absent in shh-/- mice. Based on in vitro and genetic studies, the zinc finger-containing transcription factors Gli 1, 2, and 3 are mediators of the Shh intracellular response. The floorplate and adjacent cell types are absent in gli1-/-;gli2-/- mice, but part of the Shh-/- phenotype in the neural tube is alleviated in the Shh-/-;gli3-/- double mutant. This is consistent with the predicted role of Gli3 as a repressor of the Shh response. Gli3 repressor activity is blocked by Shh. In order to test the role of the repressor form of Gli3 in the neural tube, a truncated version of Gli3 (Gli3R*) was designed to mimic a Pallister Hall allele. Gli3R* acts as a constitutive repressor independent of Shh signaling. Misexpression of Gli3R* in the chick neural tube caused a ventral expansion of class-I, dorsal progenitor proteins and a loss of class-II, ventral progenitor proteins consistent with expected activity as a repressor of the Shh response. Activation of the BMP response is sufficient to maintain gli3 expression in neural plate explants, which might be a mechanism by which BMPs antagonize the Shh response. PMID- 12729565 TI - Parp1-deficiency induces differentiation of ES cells into trophoblast derivatives. AB - Embryonic stem (ES) cells deficient in the enzyme poly(ADP-ribose) polymerase (Parp1) develop into teratocarcinoma-like tumors when injected subcutaneously into nude mice that contain cells with giant cell-like morphology. We show here that these cells express genes characteristic of trophoblast giant cells and thus belong to the trophectoderm lineage. In addition, Parp1(-/-) tumors contained other trophoblast subtypes as revealed by expression of spongiotrophoblast specific marker genes. The extent of giant cell differentiation was enhanced, however, as compared with spongiotrophoblast. A similar shift toward trophoblast giant cell differentiation was observed in cultures of Parp1-deficient ES cells and in placentae of Parp1(-/-) embryos. Analysis of other cell lineage markers demonstrated that Parp1 acts exclusively in trophoblast to suppress differentiation. Surprisingly, trophoblast derivatives were also detected in wildtype tumors and cultured ES cells, albeit at significantly lower frequency. These data show that wildtype ES cells contain a small population of cells with trophectoderm potential and that absence of Parp1 renders ES cells more susceptible to adopting a trophoblast phenotype. PMID- 12729566 TI - Myofibrillogenesis in the first cardiomyocytes formed from isolated quail precardiac mesoderm. AB - De novo assembly of myofibrils was investigated in explants of precardiac mesoderm from quail embryos to address a controversy about different models of myofibrillogenesis. The sequential expression of sarcomeric components was visualized in double- and triple-stained explants before, during, and just after the first cardiomyocytes began to beat. In explants from stage 6 embryos, cultured for 10 h, ectoderm, endoderm, and the precardiac mesoderm displayed arrays of stress fibers with alternating bands of the nonmuscle isoforms of alpha actinin and myosin IIB. With increasing time in culture, mesoderm cells contained fibrils composed of actin, nonmuscle myosin IIB, and sarcomeric alpha-actinin. Several hours later, before beating occurred, both nonmuscle and muscle myosin II localized in some of the fibrils in the cells. Concentrations of muscle myosin began as thin bundles, dispersed in the cytoplasm, often overlapping one another, and progressed to small, aligned A-band-sized aggregates. The amount of nonmuscle myosin decreased dramatically when Z-bands formed, the muscle myosin became organized into A-bands, and the cells began beating. The sequential changes in protein composition of the fibrils in the developing muscle cells supports the model of myofibrillogenesis in which assembly begins with premyofibrils and progresses through nascent myofibrils to mature myofibrils. PMID- 12729567 TI - The effect of the cyclopropyl group on the conformation of chemotactic formyl tripeptides. AB - Certain formyl peptides are powerful chemoattractants towards neutrophils. In this study, several formyl tripeptides were synthesized and used to investigate the effects of different amino acid residues in position 1 on their ability to stimulate neutrophil chemotaxis. Pig neutrophil chemotaxis towards the formyl tripeptide, HCO-Ac(3)C-Leu-Phe-OMe 1, where Ac(3)C represents 1-amino-1 cyclopropane carboxylic acid, was observed. Pig neutrophil chemotaxis towards a very similar formyl tripeptide, HCO-Aib-Leu-Phe-OMe 2, where Aib represents alpha amino isobutyric acid, was not observed. Compared to the isopropyl group, it was shown that the cyclopropyl group induces a greater percentage of the E conformation about the formamide functionality in these peptides. For 1 and 2, the E isomer distributions in CDCl3 are 36 and 9%, respectively. Since a major difference between these two peptides is the Z/E isomeric distribution, one implication is that the peptide-receptor site interactions involving the E conformer are more effective than those of the Z conformer. No pig neutrophil chemotaxis towards the formyl tripeptides, HCO-Ala-Leu-Phe-OMe 3 and HCO-Gly-Leu Phe-OMe 4 was observed. These formyl tripeptides exhibit a low percentage of the E isomer, similar to that of peptide 2. PMID- 12729569 TI - Synthesis and hydrolysis of a phenylalanyl adenylate pentacoordinated phosphorane. AB - Amino acid-nucleotide conjugates have important biological functions and therapeutic applications. For example, aminoacyl adenylates are key intermediates in aminoacyl tRNA synthetase reactions. They may also be involved in the prebiotic synthesis of polypeptides. Finally, various amino acid carbomethoxy aryl phosphoramidates of nucleotide prodrugs may be activated through a mechanism involving a pentacoordinated phosphorane intermediates. In order to understand better the chemistry of these compounds, a phenylalanyl adenylate pentacoodinated phosphorane has been synthesized in 72% yield and its decomposition in aqueous solution studied. Hydrolysis gave 2('),3(')-O-isopropylidene adenosine 5(') monophosphate, 2('),3(')-O-isopropylidene adenosine, and phenylalanine. The results provide model chemistry for the enzymatic degradation mechanism of antiviral aryl amino acid phosphodiester amidates in cells, which leads to their activation. PMID- 12729570 TI - Synthesis and tyrosinase inhibitory activity of novel N-hydroxybenzyl-N nitrosohydroxylamines. AB - Several novel N-substituted N-nitrosohydroxylamines were synthesized. They all inhibited mushroom tyrosinase, but the type of inhibition was different depending on the substituent. Some N-(mono- or dihydroxybenzyl)-N-nitrosohydroxylamines exhibited uncompetitive inhibition with respect to L-dopa. Among them, compound 6 was also a competitive inhibitor with respect to oxygen. This observation suggests that another interaction by the meta- or para-hydroxyl group might stabilize the binding of the inhibitor to the enzyme through the oxygen binding site. PMID- 12729568 TI - Synthesis and biological evaluation of the disulfide form of the glutathione analogue gamma-(L-glutamyl)-L-cysteinyl-L-aspartyl-L-cysteine. AB - By using the chain to chain mode of cyclization the title glutathione analogue (4), containing the 11-membered disulfide ring replacing the native -Cys-Gly fragment, has been synthesized and characterized together with its reduced dithiol form gamma-Glu-Cys-Asp-Cys (5). The activity of (4) with gamma-glutamyl transferase and glutathione reductase has been evaluated and compared with those of the two conformationally restricted glutathione analogues (2) and (3) previously reported. PMID- 12729571 TI - Synthesis of the minor acrolein adducts of 2(')-deoxyguanosine and their generation in oligomeric DNA. AB - Acrolein, a known mutagen, undergoes reaction in vitro under physiological conditions with both 2(')-deoxyguanosine and native DNA to give rise to exocyclic adducts of the 5,6,7,8-tetrahydropyrimido[1,2-a]purine-10(3H)-one class having an hydroxy group at either the 6 or the 8 position. Previously we have shown that the 8-hydroxy derivative in a bacterial system has very low mutagenicity probably because in double-stranded DNA this residue exists in the open-chain aldehydic form [N(2)-(3-oxopropyl)-2(')-deoxyguanosine] (3). To continue our investigation in this area, we needed ample supplies of the 6-hydroxy isomers. This current paper describes high-yield simple methods for the synthesis in bulk of the 6 hydroxy adduct 1 and its incorporation into DNA oligomers. The basic methods for the synthesis of the adduct 1, involve 1-substitution of dG derivatives with a 3 butenyl group, dihydroxylation of the olefin with osmium tetroxide and N methylmorpholine N-oxide, then diol cleavage with periodate ion after incorporation of the 1-(3,4-diacetoxybutyl)-2(')-deoxyguanosine into oligomeric DNA. PMID- 12729572 TI - Quantitative elucidation of the molecular mechanisms of hydroxyl radical quenching reactivity of phenolic compounds. AB - Reported discrepancies have confused the understanding of the molecular mechanisms of antioxidant reactivity somewhat. The consequent problems necessitate systematic investigations on the molecular orbital features of antioxidants and their correlation with antioxidant potentials. In the present work, phenolic compounds as typical antioxidants were selected to investigate their hydroxyl radical-scavenging properties, and the related mechanisms of action were studied theoretically by computational chemistry. A good correlation was observed between antioxidant activity and theoretical parameters, such as O-H bond dissociation energy (BDE), ionization potential (IP), enthalpy of electron transfer (E(a)), chemical hardness (HOMO-LUMO gap), and spin delocalization of the phenoxyl radicals (D(s)(r)). The results demonstrate that the molecular mechanisms regulating the antioxidant action were more complex than hydrogen or electron-transfer processes and explain previous contradictions. Meanwhile, a satisfactory quantitative structure-activity relationship (QSAR) model was established which should be of predictive value in evaluating or screening hydroxyl radical-scavenging antioxidants. PMID- 12729573 TI - Polysaccharide-polynucleotide complexes (15): thermal stability of schizophyllan (SPG)/poly(C) triple strands is controllable by alpha-amino acid modification. AB - Schizophyllan (SPG), a beta-1,3-glucan polysaccharide which is known to form macromolecular complexes with certain polynucleotides, was modified by a reductive amination method with alpha-amino acids (Arg, Lys, and Ser). The thermal stability of the complexes as estimated by T(m) was enhanced in SPG-Arg and SPG-Lys conjugates which have pI values higher than the pH of the medium (8.0). The T(m) shift increased with the increase in the percentage of alpha amino acid introduced and the highest T(m) values attained were 64 degrees C for SPG-Arg conjugate and 62 degrees C for SPG-Lys conjugate, which are higher by 13 and 11 degrees C, respectively, than those of the unmodified SPG+poly(C) complex. In the SPG-Ser conjugate with a pI lower than the medium pH (8.0), the T(m) values decreased with an increase in the percentage of Ser. Formation of the macromolecular complex was no longer detected above 13.2% Ser. The findings indicate that the T(m) values are easily controllable by the type and percentage of the introduced alpha-amino acids. We believe, therefore, that the present conjugates, consisting of naturally originated SPG and alpha-amino acids, provide an important lead for developing nontoxic artificial vectors and to control the affinity with polynucleotides in response to medium pH and temperature. PMID- 12729574 TI - De novo design and utilization of photolabile caged substrates as probes of hydrogen tunneling with horse liver alcohol dehydrogenase at sub-zero temperatures: a cautionary note. AB - In order to understand the influence of protein dynamics on enzyme catalysis and hydrogen tunneling, the horse liver alcohol dehydrogenase (HLADH) catalyzed oxidation of benzyl alcohol was studied at sub-zero temperatures. Previous work showed that wild type HLADH has significant kinetic complexity down to -50 degrees C due to slow binding and loss of substrate [S.-C. Tsai, J.P. Klinman, Biochemistry, 40 (2001) 2303]. A strategy was therefore undertaken to reduce kinetic complexity at sub-zero temperatures, using a photolabile (caged) benzyl alcohol that prebinds to the enzyme and yields the active substrate upon photolysis. By computer modeling, a series of caged alcohols were designed de novo, synthesized, and characterized with regard to photolysis and binding properties. The o-nitrobenzyl ether 15, with a unique long tail, was found to be most ideal. At sub-zero temperatures in 50% MeOH, a two-phase kinetic trace and a rate enhancement by the use of 15 were observed. Despite the elimination of substrate binding as a rate-limiting step, the use of caged benzyl alcohol does not produce a measurable H/D kinetic isotope effect. Unexpectedly, the observed fast phase corresponds to multiple enzyme turnovers, based on the stoichiometry of the substrate to enzyme. Possible side reactions and their effects, such as the re-oxidation of bound NADH and the dissipation of photo-excitation energy, may offer an explanation for the observed multiple-turnovers. The lack of observable deuterium isotope effects offers a cautionary note for the application of caged substrates to isolate and study chemical steps of enzyme reactions, particularly when NADH is involved in the reaction pathway. PMID- 12729575 TI - Convenient syntheses of biogenic aldehydes, 3,4-dihydroxyphenylacetaldehyde and 3,4-dihydroxyphenylglycolaldehyde. AB - The title compounds were prepared from a common precursor, a bis-THP-protected dihydroxyphenylacetic acid methyl ester. Key steps are the introduction of the alpha-hydroxyl group by Davis oxaziridine reagent and formation of the aldehydes by DIBALH ester reduction. PMID- 12729576 TI - Mitochondrial control of apoptosis: an introduction. AB - Mitochondrial membrane permeabilization [MMP] constitutes an essential step of the intrinsic pathway leading to apoptosis. Several oncoproteins, tumor suppressor gene products, viral virulence factors and pharmacological agents modulate apoptosis via direct effects on mitochondria. This has far-reaching implications for the pathophysiology of several major diseases such as cancer, neurodegeneration, and AIDS. The detailed investigation of the mechanisms of MMP will hopefully lead to the discovery of suitable drug targets for therapeutic intervention on cell death control. PMID- 12729577 TI - Mechanisms of cytochrome c release by proapoptotic BCL-2 family members. AB - A crucial amplificatory event in several apoptotic cascades is the nearly complete release of cytochrome c from mitochondria. Proteins of the BCL-2 family which include both anti- and proapoptotic members control this step. Here, we review the proposed mechanisms by which proapoptotic BCL-2 family members induce cytochrome c release. Data support a model in which the apoptotic pathway bifurcates following activation of a "BH3 only" family member. BH3 only molecules induce the activation of the multidomain proapoptotics BAX and BAK, resulting in the permeabilization of the outer mitochondrial membrane and the efflux of cytochrome c. This is coordinated with the activation of a distinct pathway characterized by profound changes of the inner mitochondrial membrane morphology and organization. This mitochondrial remodelling insures complete release of cytochrome c and the onset of mitochondrial dysfunction that is a typical feature of many apoptotic deaths. PMID- 12729578 TI - Calcium signaling and apoptosis. AB - Ca(2+) is one of the key regulators of cell survival, but Ca(2+) can also induce apoptosis in response to a variety of pathological conditions. The pro-apoptotic effects of Ca(2+) are mediated by a diverse range of Ca(2+)-sensitive factors that are compartmentalized in various intracellular organelles including the ER, cytoplasm, and mitochondria. The Ca(2+) dynamics of these organelles appear to be modulated by the apoptosis-regulating Bcl-2 family proteins. In this paper, the recent progress of research on the mechanisms mediating the apoptosis-regulating effects of Ca(2+) and the interactions of Bcl-2 family proteins with the Ca(2+) storage organelles are discussed. PMID- 12729579 TI - Mitochondrial membrane permeabilisation by Bax/Bak. AB - Recent studies on cells derived from mice deficient in both multi-domain pro apoptotic genes of the Bcl-2 family, Bax and Bak, suggest that one or other of these proteins are required for the release of apoptogens such as cytochrome c from mitochondria. In addition BH-3 only proteins of this family such as Bid are suggested to act as critical death inducing ligands via interactions with pro- and anti-apoptotic Bcl-2 family proteins with Bax or Bak at the mitochondrial surface. Despite this increase in knowledge it remains unclear precisely how Bak and Bax promote outer mitochondrial membrane (OMM) permeabilisation. We suggest that Bax and Bak may not operate in precisely the same manner and evaluate the current models for their function. We also consider the emerging information that lipid-protein interactions may be crucial to the actions of Bax and Bak. PMID- 12729580 TI - Mitochondrial permeability transition: a common pathway to necrosis and apoptosis. AB - Opening of high conductance permeability transition pores in mitochondria initiates onset of the mitochondrial permeability transition (MPT). The MPT is a causative event, leading to necrosis and apoptosis in hepatocytes after oxidative stress, Ca(2+) toxicity, and ischemia/reperfusion. CsA blocks opening of permeability transition pores and protects cell death after these stresses. In contrast to necrotic cell death which is a consequence of ATP depletion, ATP is required for the development of apoptosis. Reperfusion and the return of normal pH after ischemia initiate the MPT, but the balance between ATP depletion after the MPT and ATP generation by glycolysis determines whether the fate of cells will be apoptotic or necrotic death. Thus, the MPT is a common pathway leading to both necrotic and apoptotic cell death after ischemia/reperfusion. PMID- 12729581 TI - Redox regulation and signaling lipids in mitochondrial apoptosis. AB - Apoptosis can be regulated at multiple levels. A number of proteins with regulatory function in cell death are sensitive to cellular redox environment. The antioxidant glutathione (GSH) and redox-sensitive proteins, thioredoxin and glutathione S-transferase, thus regulate cell death pathways by modulating the redox state of specific thiol residues of target proteins including stress kinases, transcription factors, and caspases. GSH in mitochondria plays an important role in the integrity of mitochondrial proteins and lipids known to play a vital role in the permeabilization of mitochondrial membranes and release of proapoptotic factors. The regulation of mitochondrial GSH (mGSH) is determined by its uptake from the cytosol which is dependent on appropriate membrane dynamics. The deposition of cholesterol in mitochondria induced by alcohol intake impairs this translocation, resulting in severe depletion of mGSH and in sensitization to apoptosis stimuli. Although the interaction of proapoptotic proteins with mitochondria initiates apoptotic pathways, recent data indicate that the mitochondrial trafficking of glycosphingolipids, e.g., ganglioside GD3, induced by apoptotic stimuli is a key event that sets off mitochondrial-dependent apoptotic cascades. PMID- 12729582 TI - Scission, spores, and apoptosis: a proposal for the evolutionary origin of mitochondria in cell death induction. AB - Mitochondria fragment prior to caspase activation during many pathways of apoptosis. Inhibition of the machinery that normally regulates mitochondrial morphology in healthy cells inhibits the fission that occurs during apoptosis and actually delays the process of cell death. Interestingly, there are certain parallels between mitochondrial fission and bacterial sporulation. As bacterial sporulation can be considered a stress response we suggest that a primordial stress response of endosymbiont mitochondrial progenitors may have been adopted for the stress response of early eukaryotes. Thus, the mitochondrial fission process may represent an early stress response of primitive mitochondria that could have integrated the stress signals and acted as an initial sensor for the eukaryotic response system. The fact that mitochondria fragment during apoptosis using the machinery descended from or that superceded the bacterial stress response of sporulation is consistent with this hypothesis. This hypothesis would explain why what is generally considered the "power house" of the cell came to integrate the cell death response and regulate apoptosis. PMID- 12729583 TI - Mitochondrial intermembrane proteins in cell death. AB - Apoptosis is a form of programmed cell death important in the development and tissue homeostasis of multicellular organisms. Mitochondria have, next to their function in respiration, an important role in the apoptotic-signaling pathway. Malfunctioning at any level of the cell is eventually translated in the release of apoptogenic factors from the mitochondrial intermembrane space resulting in the organized demise of the cell. Some of these factors, such as AIF and endonuclease G, appear to be highly conserved during evolution. Other factors, like cytochrome c, have gained their apoptogenic function later during evolution. In this review, we focus on the role of cytochrome c, AIF, endonuclease G, Smac/DIABLO, Omi/HtrA2, Acyl-CoA-binding protein, and polypyrimidine tract binding protein in the initiation and modulation of cell death in different model organisms. These mitochondrial factors may contribute to both caspase-dependent and caspase-independent processes in apoptotic cell death. PMID- 12729584 TI - Mammalian mitochondrial IAP binding proteins. AB - Four mitochondrial proteins have been identified that immunoprecipitate with the mammalian inhibitor of apoptosis (IAP) protein XIAP. Each of them interacts via a processed amino terminus that resembles those of the insect pro-apoptotic IAP binding proteins Grim, HID, Reaper, and Sickle. Two, Diablo/Smac and HrtA2/Omi, have been extensively characterized. Both Diablo and HtrA2 can bind to IAPs and promote apoptosis when over-expressed in transfected cells, but unlike the insect IAP antagonists, to date there is scant evidence that they are important regulators of apoptosis in more physiological circumstances. PMID- 12729585 TI - Heat shock proteins, cellular chaperones that modulate mitochondrial cell death pathways. AB - Stress or heat shock proteins (HSPs) are ubiquitous and highly conserved proteins whose expression is induced in response to a wide variety of physiological and environmental insults. They allow the cells to survive to otherwise lethal conditions. Various mechanisms have been proposed to account for the cytoprotective functions of HSPs. These proteins play an essential role in intracellular "house-keeping" by assisting the correct folding of nascent and stress-accumulated misfolded proteins and preventing their aggregation. Several HSPs have also demonstrated to directly interact with various components of the tightly regulated programmed cell death machinery, upstream, and downstream of the mitochondrial events. Finally, HSPs could play a role in the proteasome mediated degradation of selected proteins under stress conditions. Altogether, these properties could make HSPs appropriate targets for modulating cell death pathways. PMID- 12729586 TI - Mitochondria at the heart of the cytotoxic attack. AB - Granzyme B is a serine proteinase that acts as a key effector of cell death mediated by cytotoxic T lymphocytes. The enzyme is transferred from the cytotoxic cell to the pathogenic target cell where it cleaves and activates a number of substrates involved in the induction of apoptosis. However, recent evidence implicates mitochondria as playing an important role in both the initiation of apoptosis and control of substrate cleavage by granzyme B in cytotoxic T lymphocyte induced death. This review focuses on current research in this rapidly expanding field, specifically the role of mitochondria in cell death induced by components of cytotoxic granules in particular granzyme B. PMID- 12729587 TI - MtDNA mutations in aging and apoptosis. AB - There is considerable evidence that the oxidative phosphorylation capacity of human mitochondria declines in various tissues with aging. However, the genetic basis of this phenomenon has not yet been clarified. The occurrence of large deletions in mtDNA from brain, skeletal, and heart muscles and other tissues of old subjects at relatively low levels has been well documented. We discuss their possible functional relevance for the aging processes. On the contrary, until very recently, only inconclusive and often discordant evidence was available for the accumulation of mtDNA point mutations in old individuals. In the past few years, however, an aging-dependent large accumulation of mtDNA point mutations has been demonstrated in the majority of individuals above a certain age. These mutations occur in the mtDNA main control region at critical sites for mtDNA replication in fibroblasts and skeletal muscles. The extraordinary tissue specificity and nucleotide selectivity of these mutations strongly support the idea of their being functionally relevant. Evidence in agreement with this conclusion has been provided by the very recent observation that an mtDNA mutation occurring in blood leukocytes near an origin of replication, which causes a remodeling of this origin, occurs at a strikingly higher frequency in centenarians and monozygotic and dizygotic twins than in the control populations, strongly pointing to its survival value. The present article reviews another area of active research and discussion, namely, the role of pathogenic mtDNA mutations in causing programmed cell death. The available evidence has clearly shown that mtDNA and respiration are not essential for the process of apoptosis. However, the limited and sometimes contradictory data indicate that the absence or impaired function of mtDNA can influence the rate of this process, most probably by regulating the production of reactive oxygen species or the lack thereof. PMID- 12729588 TI - Mitochondrial apoptotic pathways induced by Drosophila programmed cell death regulators. AB - Multicellular organisms eliminate unwanted or damaged cells by cell death, a process essential to the maintenance of tissue homeostasis. Cell death is a tightly regulated event, whose alteration by excess or defect is involved in the pathogenesis of many diseases such as cancer, autoimmune syndromes, and neurodegenerative processes. Studies in model organisms, especially in the nematode Caenorhabditis elegans, have been crucial in identifying the key molecules implicated in the regulation and execution of programmed cell death. In contrast, the study of cell death in Drosophila melanogaster, often an excellent model organism, has identified regulators and mechanisms not obviously conserved in other metazoans. Recent molecular and cellular analyses suggest, however, that the mechanisms of action of the main programmed cell death regulators in Drosophila include a canonical mitochondrial pathway. PMID- 12729589 TI - Mitochondrial potassium channels and uncoupling proteins in synaptic plasticity and neuronal cell death. AB - The function of the nervous system relies upon synaptic transmission, a process in which a neurotransmitter released from pre-synaptic terminals of one neuron (in response to membrane depolarization and calcium influx) activates post synaptic receptors on dendrites of another neuron. Synapses are subjected to repeated bouts of oxidative and metabolic stress as the result of changing ion gradients and ATP usage. Mitochondria play central roles in meeting the demands of synapses for ATP and in regulating calcium homeostasis, and mitochondrial dysfunction can cause dysfunction and degeneration of synapses, and can trigger cell death. We have identified two types of mitochondrial proteins that serve the function of protecting synapses and neurons against dysfunction and death. Mitochondrial ATP-sensitive potassium (MitoKATP) channels modulate inner membrane potential and oxyradical production; mitochondrial potassium fluxes can affect cytochrome c release and caspase activation and may determine whether neurons live or die in experimental models of stroke and Alzheimer's disease. Uncoupling proteins (UCPs) are a family of mitochondrial membrane proteins that uncouple electron transport from ATP production by transporting protons across the inner membrane. Neurons express at least three UCPs including the widely expressed UCP 2 and the neuron-specific UCP-4 and UCP-5 (BMCP-1). We have found that UCP-4 protects neurons against apoptosis by a mechanism involving suppression of oxyradical production and stabilization of cellular calcium homeostasis. The expression of UCP-4 is itself regulated by changes in energy metabolism. In addition to their roles in neuronal cell survival and death, MitoKATP channels and UCPs may play roles in regulating neuronal differentiation during development and synaptic plasticity in the adult. PMID- 12729590 TI - Mitochondria and ischemic reperfusion damage in the adult and in the developing brain. AB - The developing and the adult brain respond in similar ways to ischemia, but also display clear differences. For example, the relative contributions of necrosis and apoptosis to neuronal death may be different, such that apoptotic mechanisms would be more prevalent in the developing brain. During normal development, more than half of the neurons in some brain regions are removed through apoptosis, and effectors like caspase-3 are highly upregulated in the immature brain. Mitochondria are pivotal regulators of cell death through their role in energy production and calcium homeostasis, their capacity to release apoptogenic proteins and to produce reactive oxygen species. This review will summarize some of the current studies dealing with mitochondria-related mechanisms of ischemic brain damage, with special reference to developmental aspects. PMID- 12729591 TI - Mitochondria in HIV-1-induced apoptosis. AB - It is now well admitted that HIV infection leading to AIDS is associated with an abnormal susceptibility of T cells to undergo apoptosis. Recent progress in research into programmed cell death has resulted in the identification of the principal pathways involved in this process. Thus the "extrinsic" as well as the "intrinsic" pathways converge to the mitochondria considered as the main sensor of programmed cell death. This review summarizes our knowledge of the influence of mitochondrial control on T cell death during HIV and SIV infections. PMID- 12729592 TI - Mitochondrion-targeted apoptosis regulators of viral origin. AB - During coevolution with their hosts, viruses have "learned" to intercept or to activate the principal signal transducing pathways leading to cell death. A number of proteins from pathophysiologically relevant viruses are targeted to mitochondria and regulate (induce or inhibit) the apoptosis-associated permeabilization of mitochondrial membranes. Such proteins are encoded by human immunodeficiency virus 1, Kaposi's sarcoma-associated herpesvirus, human T-cell leukemia virus-1, hepatitis B virus, cytomegalovirus, and Epstein Barr virus, among others. Within mitochondria, such apoptosis regulators from viral origin can target distinct proteins from the Bcl-2 family and the permeability transition pore complex including the adenine nucleotide translocase, cyclophilin D, the voltage-dependent anion channel, and the peripheral benzodiazepine receptor. Thus, viral proteins can regulate apoptosis at the mitochondrial level by acting on a variety of different targets. PMID- 12729593 TI - Mechanism of HIV-1 viral protein R-induced apoptosis. AB - The paradigm of HIV-1 infection includes the diminution of CD4(+) T cells, loss of immune function, and eventual progression to AIDS. However, the mechanisms that drive host T cell depletion remain elusive. One HIV protein thought to participate in this destructive cascade is the Vpr gene product. Accordingly, we review the biology of the HIV-1 viral protein R (Vpr) an apoptogenic HIV-1 accessory protein that is packaged into the virus particle. In this review we focus specifically on Vpr's ability to induce host cell apoptosis. Recent evidence suggests that Vpr implements a unique mechanism to drive host cell apoptosis, by directly depolarizing the mitochondria membrane potential. Vpr's attack on the mitochondria results in release of cytochrome c resulting in activation of the caspase 9 pathway culminating in the activation of caspase 3 and the downstream events of apoptosis. Vpr may interact with the adenine nucleotide translocator (ANT) to prompt this cascade. The role of Vpr-induced apoptosis in HIV pathogenesis is considered. PMID- 12729594 TI - Development of a fluorescent probe for the study of nucleosome assembly and dynamics. AB - To develop a probe for use in real-time dynamic studies of nucleosomes, core histones (from Drosophila) were conjugated to a DNA-intercalating dye, thiazole orange, by a reaction targeting Cys 110 of histone H3. In the absence of DNA, the conjugated histones are only very weakly fluorescent. However, upon reconstitution into nucleosomes by standard salt dialysis procedures, the probe fluoresces strongly, reflecting its ability to intercalate into the nucleosomal DNA. The probe is also sensitive to the nature of the DNA-histone interaction. Nucleosomes reconstituted by stepwise salt dialysis give a fluorescence signal quite different from that of the species formed when DNA and histones are simply mixed in low salt. In addition, changing either the DNA length or the type of sequence (nucleosome positioning sequences versus random DNA of the same size) used in the reconstitution alters the resulting fluorescence yield. The results are all consistent with the conclusion that a more rigid, less flexible nucleosome structure results in less fluorescence than a looser structure, presumably due to structural constraints on dye intercalation. This probe should be well suited to analyzing nucleosome dynamics and to following factor-mediated assembly and remodeling of nucleosomes in real time, particularly at the single molecule level. PMID- 12729595 TI - Phosphomannomutase activity in congenital disorders of glycosylation type Ia determined by direct analysis of the interconversion of mannose-1-phosphate to mannose-6-phosphate by high-pH anion-exchange chromatography with pulsed amperometric detection. AB - Congenital disorders of glycosylation (CDG) are a group of multisystemic disorders resulting from defects in the synthesis and processing of N-linked oligosaccharides. The most common form, CDG type Ia (CDG-Ia), results from a deficiency of the enzyme phosphomannomutase (PMM). PMM converts mannose 6 phosphate (man-6-P) to mannose-1-phosphate (man-1-P), which is required for the synthesis of GDP-mannose, a substrate for dolichol-linked oligosaccharide synthesis. The traditional assay for PMM, a coupled enzyme system based on the reduction of NADP(+) to NADPH using man-1-P as a substrate, has limitations in accuracy and reproducibility. Therefore, a more sensitive, direct test for PMM activity, based on the detection of the conversion of man-1-P to man-6-P by high pH anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD), was developed. Using this assay, the activity of PMM was markedly deficient in fibroblasts and lymphoblasts from 23 patients with CDG-Ia (range 0-15.3% of control, average 4.9+/-4.7%) and also decreased in seven obligate heterozygotes (range 33.0-72.0% of control, average 52.2+/-14.7%). Unlike the spectrophotometric method, there was no overlap in PMM activity among patients, obligate heterozygotes, or controls. Thus, the PMM assay based on HPAEC-PAD has increased utility in the clinical setting, and can be used, together with transferrin isoelectric focusing, to diagnose patients with CDG-Ia and to identify heterozygotes when clinically indicated. PMID- 12729598 TI - Electrochemical detection of protein-protein interactions using a yeast two hybrid: 17-beta-estradiol as a model. AB - In this work we present a modified yeast two-hybrid bioassay for the highly sensitive detection of protein-protein interactions, based on the electrochemical monitoring of beta-D-galactosidase reporter gene activity, using p-aminophenyl beta-D-galactopyranoside (PAPG) as a synthetic substrate. In a model system, the sensitive detection of 17-beta-estradiol was achieved at concentrations as low as 10(-11)M (approx 2 pg/ml) by monitoring 17-beta-estradiol receptor dimerization after exposure to 17-beta-estradiol. The sensitivity of this system was higher than that of standard optical methods by three orders of magnitude. PMID- 12729596 TI - Assessment of cell concentration and viability of isolated hepatocytes using flow cytometry. AB - The assessment of cell concentration and viability of freshly isolated hepatocyte preparations has been traditionally performed using manual counting with a Neubauer counting chamber and staining for trypan blue exclusion. Despite the simple and rapid nature of this assessment, concerns about the accuracy of these methods exist. Simple flow cytometry techniques which determine cell concentration and viability are available yet surprisingly have not been extensively used or validated with isolated hepatocyte preparations. We therefore investigated the use of flow cytometry using TRUCOUNT Tubes and propidium iodide staining to measure cell concentration and viability of isolated rat hepatocytes in suspension. Analysis using TRUCOUNT Tubes provided more accurate and reproducible measurement of cell concentration than manual cell counting. Hepatocyte viability, assessed using propidium iodide, correlated more closely than did trypan blue exclusion with all indicators of hepatocyte integrity and function measured (lactate dehydrogenase leakage, cytochrome p450 content, cellular ATP concentration, ammonia and lactate removal, urea and albumin synthesis). We conclude that flow cytometry techniques can be used to measure cell concentration and viability of isolated hepatocyte preparations. The techniques are simple, rapid, and more accurate than manual cell counting and trypan blue staining and the results are not affected by protein-containing media. PMID- 12729597 TI - Stable isotope phospho-profiling of fibrinogen and fetuin subunits by element mass spectrometry coupled to capillary liquid chromatography. AB - We have used one-dimensional polyacrylamide gel electrophoresis, tryptic digestion, and capillary liquid chromatography-mass spectrometry with inductively coupled plasma ionization and phosphorus-31 detection or electrospray ionization for the analysis of protein phosphorylation. We have analyzed human fibrinogen with two well-characterized phosphorylation sites and bovine fetuin with unknown phosphorylation status. Both serine-3 and serine-345 (both in Aalpha) of fibrinogen were clearly recognized. In bovine fetuin, four phosphorylated sites were newly characterized (serine-138, serine-320, serine-323, and serine-324). The novel strategy provides a fast and quantitative overview of the presence of protein phosphorylation sites. PMID- 12729599 TI - Filter binding assay for the geldanamycin-heat shock protein 90 interaction. AB - A filter binding assay to measure affinity of [3H-allyl]17-allylamino geldanamycin ([3H]AAG) for the ATP binding site of the N-terminal domain of human Hsp90alpha (hHsp90alpha9-236) was developed. Diethylaminoethyl cellulose or glass fiber filters impregnated with polyethyleneimine were used to capture the [3H]AAG Hsp90 complex, and conditions which washed >98% of free [3H]AAG from the filters were developed. The complex formed at a rapid rate (k(on)=2.5 x 10(7)Lmol(-1) x s(-1)) and dissociated with a half-life of 2.3 min (k(off)=5 x 10(-3) x s(-1)). hHsp90alpha9-236 bound to [3H]AAG with a K(d) value of 0.4+/-0.1 microM. [3H]AAG had similar affinities for full-length hHsp90alpha and for hHsp90alpha9-236 variants containing biotinylated N-terminal biotinylation signal sequences and N- or C-terminal His(6) tags. Geldanamycin, ADP, ATP, and radicicol-all known to bind to the ATP domain of Hsp90-competed with [3H]AAG for binding to hHsp90alpha9 236, showing K(d) values in good agreement with reported values. PMID- 12729600 TI - Characterization of a series of far-red-absorbing thiobarbituric acid oxonol derivatives as fluorescent probes for biological applications. AB - Chromophores that absorb in the far-red region of the spectrum are increasingly being utilized for applications in the biosciences. We have synthesized and evaluated a novel series of fluorescent oxonols based on thiobarbituric acids containing aryl and heteroaryl substituents. The novel chromophores possess narrow absorption spectra ( approximately 40-nm bandwidths), reasonable Stokes shifts ( approximately 25 nm), and quantum yields of up to 0.67 in organic solvents and 0.3 in aqueous solvents, with absorption wavelength maxima at 620 640 nm. The spectral properties of the compounds are sensitive to base and exhibit a loss of far-red absorbance that is concentration and time dependent. Derivatives have been synthesized that can be used for the labeling of macromolecules such as proteins and DNA. The probes show environment sensitivity and the oligonucleotide conjugates sense the formation of duplex DNA. These novel far-red fluorophores have potential applications in diagnostic and research applications. PMID- 12729601 TI - The effects of Ca(2+) binding on the conformation of calbindin D(28K): a nuclear magnetic resonance and microelectrospray mass spectrometry study. AB - Calbindin D(28K) is a six-EF-hand calcium-binding protein found in the brain, peripheral nervous system, kidney, and intestine. There is a paucity of information on the effects of calcium binding on calbindin D(28K) structure. To further examine the mechanism and structural consequences of calcium binding to calbindin D(28K) we performed detailed complementary heteronuclear NMR and microelectrospray mass spectrometry investigations of the calcium-induced conformational changes of calbindin D(28K). The combined use of these two powerful analytical techniques clearly and very rapidly demonstrates the following: (i). apo-calbindin D(28K) has an ordered structure which changes to a notably different ordered conformation upon Ca(2+) loading, (ii). calcium binding is a sequential process and not a simultaneous event, and (iii). EF-hands 1, 3, 4, and 5 take up Ca(2+), whereas EF-hands 2 and 6 do not. Our results support the opinion that calbindin D(28K) has characteristics of both a calcium sensor and a buffer. PMID- 12729602 TI - Quantitation of cytochrome c release from rat liver mitochondria. AB - The apoptogenic protein cytochrome c can be quantitated by reverse-phase HPLC, but this method is not utilized by those who investigate mechanisms of cell death. Here, we extend the sensitivity of the method to exceed that available from immunogenic approaches and report specific procedures for applying the method to preparations of intact mitochondria, and to supernatants and pellets that arise from mitochondrial incubations. The detection limit corresponds to 0.6% of total cytochrome c found in 100 microg of rat liver mitochondrial protein, or to all of the cytochrome c that is expected in approximately 6000 hepatocytes. A single determination can be completed in 20 min, compared to a time scale of days for Western blotting methods, or hours for ELISA-based methods. The procedures are illustrated by experiments that determine the amount of cytochrome c released following the mitochondrial permeability transition as a function of medium ionic strength, and by long-term incubations of intact mitochondria in the presence and absence of an exogenous oxidizable substrate. Swelling and the release of adenylate kinase activity have been determined simultaneously to show how the data can be applied to evaluate the role of outer membrane disruption in mechanisms that release cytochrome c. PMID- 12729603 TI - DNA probe attachment on plastic surfaces and microfluidic hybridization array channel devices with sample oscillation. AB - DNA probe immobilization on plastic surfaces and device assembly are both critical to the fabrication of microfluidic hybridization array channel (MHAC) devices. Three oligonucleotide (oligo) probe immobilization procedures were investigated for attaching oligo probes on four different types of plastic surfaces (polystyrene, polycarbonate, poly(methylmethacrylate), and polypropylene). These procedures are the Surmodics procedure, the cetyltrimethylammonium bromide (CTAB) procedure, and the Reacti-Bind procedure. To determine the optimal plastic substrate and attachment chemistry for array fabrication, we investigated plastic hydrophobicity, intrinsic fluorescence, and oligo attachment efficiency. The Reacti-Bind procedure is least effective for attaching oligo probes in the microarray format. The CTAB procedure performs well enough to use in array fabrication, and the concentration of CTAB has a significant effect on oligo immobilization efficiency. We also found that use of amine-modified oligo probes resulted in better immobilization efficiency than use of unmodified oligos with the CTAB procedure. The oligo probe immobilization on plastic surfaces by the Surmodics procedure is the most effective with regard to probe spot quality and hybridization sensitivity. A DNA hybridization assay on such a device results in a limit of detection of 12pM. Utilizing a CO(2) IR laser machining and adhesive layer approach, we have developed an improved procedure for realizing a DNA microarray inside a microfluidic channel. This device fabrication procedure allows for more feasible spot placement in the channel and reduced sample adsorption by adhesive tapes used in the fabrication procedure. We also demonstrated improved hybridization kinetics and increased detection sensitivity in MHAC devices by implementing sample oscillation inside the channel. A limit of detection of 5pM has been achieved in MHAC devices with sample oscillation. PMID- 12729604 TI - Separation and quantification of cellulases and hemicellulases by capillary electrophoresis. AB - Cellulases and hemicellulases are two classes of enzymes produced by filamentous fungi and secreted into the cultivation medium. Both classes of enzymes consist of a subset of classes of which the fungi produce several enzymes with varying molecular mass and pI but similar enzymatic activities. Current methods are limited in their ability to quantify all of these enzymes when all are present simultaneously in a mixture. Five different cellulases (two cellobiohydrolases and three endoglucanases) and one hemicellulase (endoxylanase) were separated using capillary electrophoresis (CE) in a fused silica capillary at pH values close to neutral. The improvement of the separation of these six proteins by the addition of alpha,omega-diaminoalkanes with chain lengths from three to seven carbon units was investigated. Dynamically coating the capillary with 1,3 diaminopropane resulted in separation of the six enzymes and the reproducibility of the migration times was between 0.6 and 1.9%. Two cases-quantitative determination of the enzyme concentrations in cultivation samples and investigation of adsorption of the enzymes onto cellulose-demonstrated the advantages and perspectives of CE analysis of these broad groups of enzymes. PMID- 12729605 TI - Miniaturizable homogenous time-resolved fluorescence assay for carboxypeptidase B activity. AB - An epitope-unmasking, homogeneous time-resolved fluorescence (HTRF) assay has been developed for measuring carboxypeptidase B (CPB) activity in a miniaturized high-throughput screening format. The enzyme substrate (biotin-RYRGLMVGGVVR-OH) is cleaved by CPB at the C terminus, causing release of the C-terminal Arg residue. The product (biotin-RYRGLMVGGVV-OH) is recognized specifically by a monoclonal antibody (G2-10) which is labeled with Eu(3+)-cryptate ([Eu(3+)]G2-10 mAb), and the complex is detected by fluorescence resonance energy transfer using streptavidin labeled with allophycocyanin ([XL665]SA). The CPB HTRF assay is readily adapted from 96- to 1536-well format as a robust (Z(')>0.5) assay for high-throughput screening. PMID- 12729607 TI - General properties of GFP-display, an electrophoretic analysis for single amino acid changes in target polypeptides. AB - The migrating position of green fluorescent protein (GFP)-fused polypeptide varied on an SDS/urea gel by a single amino acid change in the fused polypeptide segment. An easy detection method for a single amino acid change based on this observation was called "GFP-display." Using various target polypeptides, staphylococcal protein A (SpA), Ras, p53, and human beta3 adrenergic receptor (AR), and their mobility-shift patterns resulting from the single amino acid changes, several important properties of GFP-display were revealed as follows: (i). since the binding of dodecyl sulfate ions to acidic or hydrophilic amino acids is weaker than that to basic or hydrophobic amino acids, the ions bound weakly to the fused polypeptide segment are forced to come off by high concentrations of urea prior to the ions bound strongly, resulting in the mobility shift, (ii). the mobility shift is estimated to a certain extent using a new parameter called the "GD value" calculated from the isoelectric point, hydrophilicity, and number of fused amino acids, and (iii). the fluorescence intensity of GFP-fused polypeptide tends to increase with the average hydrophilicity of the fused polypeptide segment. GFP-display will be a helpful technique for many kinds of gene or protein studies related to amino acid substitutions such as the random mutagenesis in a gene of interest. PMID- 12729606 TI - Synthesis of acylglucuronides of drugs using immobilized dog liver microsomes octadecylsilica particles coated with phospholipid. AB - Immobilized dog liver microsome octadecylsilica (ODS) particles coated with phospholipid were developed for the synthesis of acylglucuronides of drugs. The phospholipid-coated ODS particles were readily prepared by stirring a solution containing L-alpha-dipalmitoylphosphatidylcholine with the ODS particles, in which the phospholipid was absorbed on the ODS surfaces by hydrophobic interaction between the acyl group of phospholipid and the otcadecyl group of the ODS particles. Similarly, the microsome-immobilized particles were readily prepared by stirring a buffer solution containing dog liver microsomes with the phospholipid-coated ODS particles, in which the microsomes were immobilized on the phospholipid-coated ODS particles by hydrophobic binding. The microsome immobilized particles exhibited UDP-glucuronosyltransferase activity which catalyzed the glucuronidation of ketoprofen and a nonpeptide endothelin receptor antagonist, S-1255 ([R]-[+]-2-[benzo(1,3)dioxol-5-yl]-6-isopropyl-4-[4 methoxyphenyl]-2H-chromene-3-carboxylic acid), to the corresponding acylglucuronide in the presence of uridine 5(')-diphosphate (UDP)-glucuronic acid, and two acylglucuronides of ketoprofen and S-1255 were synthesized using the microsome-immobilized particles. These acylglucuronides were synthesized by simply shaking the microsome-immobilized particles adsorbed on the substrate in a buffer solution containing UDP-glucuronic acid with a thermostated mixer. The molecular weights and chemical structures of the synthesized acylglucuronides were identified by mass spectrometry and nuclear magnetic resonance, respectively. The productivity of S-1255 acylglucuronide using microsome immobilized particles was approximately threefold higher than that observed with free microsomes, whereas the ketoprofen acylglucuronide productivity was slightly lower than that observed with free microsomes. The present method should be very useful for the synthesis of acylglucuronides of drugs, which are slightly soluble aqueous solutions in the drug development stage. PMID- 12729608 TI - A novel protein-based heme sensor consisting of green fluorescent protein and apocytochrome b(562). PMID- 12729609 TI - Detection and quantification of transcripts for the catalytic subunit TERT and the RNA component of telomerase in rat tissue. PMID- 12729610 TI - In situ electroporation of large numbers of cells using minimal volumes of material. PMID- 12729611 TI - The role of NFAT5/TonEBP in establishing an optimal intracellular environment. AB - NFAT5/TonEBP, the most recently described member of the rel/NFkappaB/NFAT family of signal-dependent transcription factors, is activated by extracellular hypertonicity-a cellular stress of particular and perhaps unique physiologic relevance to cells of the renal medulla. Accumulating evidence suggests that NFAT5/TonEBP also functions in vivo under isotonic conditions as part of a ubiquitous regulatory mechanism that senses and adjusts available intracellular volume during cell growth to establish an intracellular environment appropriate for optimal cell proliferation. PMID- 12729612 TI - Carboxypeptidase A-catalyzed direct conversion of leukotriene C4 to leukotriene F4. AB - Leukotrienes (LTs) are 5-lipoxygenase (5-LO)-derived arachidonic metabolites that constitute a potent set of lipid mediators produced by inflammatory cells. Leukotriene A(4), a labile allylic epoxide formed from arachidonic acid by dual 5 LO activity, is the precursor for LTB(4) and LTC(4) synthesis. LTC(4) is further transformed enzymatically by the sequential action of gamma glutamyltranspeptidase and dipeptidase to LTD(4) and LTE(4), respectively. In this report, we present evidence that bovine pancreatic carboxypeptidase A (CPA), which shares significant sequence homology with CPA in mast cell granules, catalyzes the conversion of LTC(4) to LTF(4) via the hydrolysis of an amide bond. The identity of CPA-catalyzed LTC(4) hydrolysis product as LTF(4) was confirmed by several analytical criteria, including enzymatic conversion to conjugated tetraene by soybean LO, conversion to LTE(4) by gamma-glutamyltranspeptidase, cochromatography with the standard LTF(4) and positive-ion fast-atom bombardment mass spectral analysis. Thus, it appears that the physiological significance of this single-step transformation may point toward a major cellular homeostatic mechanism of metabolizing LTC(4), a potent bronco- and vasoconstrictor, to a less potent form of cysteinyl LTs. PMID- 12729614 TI - Contact sites from human placental mitochondria: characterization and role in progesterone synthesis. AB - To understand the functional compartmentalization of human placental mitochondria, we analyzed the composition and steroidogenic activity of contact sites. Several fractions containing contact sites were isolated using osmotic shock treatment and sucrose gradient centrifugation. These fractions contained various proteins and marker enzymes associated with mitochondrial membranes. The fractions containing the cytochrome P450 side chain cleavage system, cholesterol, nicotinamide adenine dinucleotide phosphate-isocitrate dehydrogenase, porin, and adenosine 5(')-triphosphate-diphosphohydrolase activity showed the capacity to synthesize progesterone. Our observations indicate that all necessary elements and enzymes for steroidogenesis are present and functional in placental mitochondrial contact sites. This organization may facilitate the metabolism of cholesterol delivered to the outer mitochondrial membrane into steroid hormones by the inner mitochondrial membrane cholesterol side chain cleavage system. PMID- 12729615 TI - Proteoglycans synthesized and secreted by pancreatic islet beta-cells bind amylin. AB - Pancreatic islet amyloid deposits in type 2 diabetes are associated with decreased islet beta-cell function. They contain both amylin (islet amyloid polypeptide), the beta-cell-derived unique fibrillogenic component, and heparan sulfate proteoglycans (HSPGs). We hypothesized that beta-cell HSPGs contribute to islet amyloidogenesis. [35S]Sulfate-labeled proteoglycans from islet-derived beta TC3 cell cultures eluted from diethylaminoethyl Sephacel at 0.35M NaCl. Chromatography on Sepharose CL-4B and SDS-PAGE analysis revealed distinct populations of proteoglycans. Medium HSPGs eluted at K(av) approximately 0.18 and 0.50 with glycosaminoglycan chains of approximately 28 and 19 kDa, respectively. A third population containing chondroitin/dermatan sulfate eluted at K(av) approximately 0.70 with glycosaminoglycan chains of approximately 10 kDa. A single size class of heparan and chondroitin/dermatan sulfate proteoglycans in the cell layer eluted at K(av) approximately 0.40 with glycosaminoglycan chains of approximately 19 kDa. Medium and cell layer proteoglycans bound exclusively to fibrillogenic amylin, as determined by gel mobility shift assays, indicating a possible role for beta-cell-derived proteoglycans in islet amyloid formation. PMID- 12729613 TI - Involvement of the electrophile responsive element and p53 in the activation of hepatic stellate cells as a response to electrophile menadione. AB - The cytotoxic effects of menadione and hydrogen peroxide were examined in two hepatic stellate cell lines derived from normal or cirrhotic rat liver. The cirrhotic fat-storing cells (CFSC) were found more resistant than the normal fat storing cells (NFSC) to menadione cytotoxicity. No significant differences were observed in hydrogen peroxide toxicity in these two cell lines. Although protein levels and enzymatic activities of catalase, Cu,Zn-SOD, Mn-SOD, and NADPH cytochrome c reductase were similar in these cell lines, 20-fold increases of NAD(P)H:quinone oxidoreductase 1 (NQO1) enzymatic activity and protein levels were detected in CFSC compared to those of NFSC. Gel mobility shift assays and functional analysis using transient transfection experiments indicated the involvement of the electrophile responsive element (EPRE) in the up-regulation of the NQO1 expression. Antibody supershift analysis revealed that, although Nrf2 is a member of the EPRE-binding complex in both NFSC and CFSC, Nrf1 was identified as a part of the protein/DNA complex only in CFSC. Expression of p53 tumor suppressor gene was found in higher levels in CFSC than in NFSC. We conclude that activation of the EPRE-signaling pathway, which up-regulates several phase II genes and affects p53 stabilization, may offer resistance to hepatic stellate cells against oxidative damage during hepatic injury. This resistance may be a part of the activation process of the hepatic stellate cells and could contribute to their increased proliferation and production of extracellular matrix. PMID- 12729616 TI - How do calcium ions induce free radical oxidation of hydroxy-1,4-naphthoquinone? Ca2+ stabilizes the naphthosemiquinone anion-radical of echinochrome A. AB - A surprising effect is the direct action of Ca(2+) on redox reactions of ortho quinoid compounds. The effect of Ca(2+) on oxidation of the sea urchin pigment 6 ethyl-2,3,5,7,8-pentahydroxy-1,4-naphthoquinone (echinochrome A) has been studied by electron paramagnetic resonance (EPR) spectroscopy, by UV/VIS absorbance spectroscopy, and by measurement of oxygen consumption. Echinochrome A per se reacted with dioxygen only in an alkaline solution; 2,3-semiquinone anion-radical of echinochrome A and superoxide anion-radical were the intermediates of the oxidation. Addition of calcium ions sharply increased the rate of echinochrome A autooxidation at alkaline pH and provoked oxidation at neutral pH. To explain this phenomenon we have focused on changes of the acid-base properties of echinochrome A in the presence of calcium and on stabilization of 2,3-semiquinone anion-radical of echinochrome A by Ca(2+). Dissociation constants (pK(a1), pK(a2), and pK(a3)) of echinochrome A determined by potentiometric titration were 5.20, 6.78, and >10 in calcium-free solution and 5.00, 6.10, and 7.15 in the presence of Ca(2+). We have found that Ca(2+) forms an insoluble adduct with the 2,3-semiquinone anion-radical. Thus, the effect of redox-inert calcium on the free radical reactions could be explained (i) by additional deprotonation of echinochrome A and (ii) by formation of a Ca(2+)-naphtho-2,3-semiquinone complex (calcium semiquinonate). Additionally, we have shown that the dried red spines from Strongylocentrotus intermedius possess paramagnetic properties; the EPR signal of the natural spines was similar to that of calcium semiquinonate obtained in our artificial chemical system. PMID- 12729617 TI - Trifluoroethanol-induced "molten globule" state in stem bromelain. AB - 2,2,2-Trifluoroethanol (TFE) denatures proteins but also stabilizes/induces alpha helical conformation in partially/completely unfolded proteins. As reported earlier from this laboratory, stem bromelain is known to exist as a partially folded intermediate (PFI) at pH 2.0. The effect of increasing concentration of TFE on the PFI of bromelain has been investigated by circular dichroism (CD), fluorescence emission spectroscopy, binding of the hydrophobic dye 1-anilino 8 naphthalene sulfonic acid (ANS), and near-UV CD temperature transition. Far-UV CD spectra show considerable accumulation of secondary structure at 70% (v/v) concentration of TFE with spectral features resembling the pH 7.0 preparation. Interestingly the partially folded intermediate regained significant tertiary structure/interactions, with increasing concentration of TFE, and at 60% (v/v) TFE approached almost that of the pseudo native (pH 7.0) state. Further increase to 70% (v/v) TFE, however, resulted in complete loss of tertiary structure/interactions. Studies on tryptophan fluorescence also suggested the induction of some compact structure at 60% (v/v) concentration of TFE. The partially folded intermediate showed enhanced binding of the fluorescent probe (ANS) in the presence of 60% (v/v) TFE. Taken together these observations suggest a "molten globule" state between 60 and 70% (v/v) TFE. Thermal transition studies in the near-UV CD region indicated cooperative transition for PFI in the presence of 60% (v/v) TFE changing to noncooperative transition at 70% (v/v) TFE. This was accompanied by a shift in the midpoint of thermal denaturation (T(m)) from 58 to 51 degrees C. Gradual transition and loss of cooperative thermal unfolding in the 60-70% (v/v) range of TFE also support the existence of the molten globule state. PMID- 12729618 TI - Enhanced phosphorylation and enzymatic activity of phosphoglucomutase by the Btk29A tyrosine kinase in Drosophila. AB - The Drosophila Btk29A tyrosine kinase is suggested to be involved in diverse processes, although its target proteins are unknown. In the present study, we investigated substrates of Btk29A tyrosine kinase by expressing a catalytically activated form of Btk29A-P1 (Btk-EG) in Drosophila compound eyes. Expression in eye disks led to the development of the rough-eye phenotype and increased tyrosine phosphorylation of a 65-kDa protein. Partial amino acid sequence analysis of this protein showed that it was phosphoglucomutase. Phosphoglucomutase activity in heads from Btk-EG-expressing flies was higher than that in controls, suggesting that the levels of tyrosine phosphorylation and activity of the enzyme are associated with Btk29A tyrosine kinase activity. PMID- 12729619 TI - Cadmium induces impaired glucose tolerance in rat by down-regulating GLUT4 expression in adipocytes. AB - Cadmium (Cd) has been known to cause hyperglycemia with diabetes-related complications in experimental animals; however, the molecular basis underlying this Cd-induced hyperglycemia is not known. Here, we report the novel finding that the impaired glucose tolerance (IGT) in rats induced by CdCl(2) is accompanied by a drastic (by as much as 90%) and dose-dependent reduction in GLUT4 protein and GLUT4 mRNA levels in adipocytes. The effect was specific to GLUT4; neither GLUT1 nor insulin-responsive aminopeptidase in adipocytes was affected. GLUT2 in hepatocytes was also not affected. Interestingly, the effect on GLUT4 was also specific to adipocytes; the muscle tissues of the Cd-treated rats showed only a slight (<25%) reduction in GLUT4 protein level with no change in GLUT4 message level, and again with no change in GLUT1 protein and its message levels. Although the insulin-induced GLUT4 translocation in adipocytes was not affected by the Cd treatment, the 3-O-methy-D-glucose flux in insulin-stimulated adipocytes of Cd-treated rat was drastically reduced. Together these findings clearly demonstrate that Cd induces IGT in rats by selectively down-regulating GLUT4 expression in adipocytes. PMID- 12729620 TI - Alkaline treatment has contrasting effects on the structure of deglycosylated and glycosylated forms of glucose oxidase. AB - X-ray crystallographic studies on glucose oxidase showed a strong interaction between carbohydrate and protein moieties of the glycoprotein. However, experimental studies under physiological conditions reported no influence of carbohydrate moiety on the structural and functional properties of glucose oxidase. In order to demonstrate the role of carbohydrate moiety on the structure and stability, we carried out a detailed comparative study on the pH-induced structural changes in the native and deglycosylated forms of glucose oxidase. Our studies demonstrate that at physiological pH both forms of enzyme have very similar structural and stability properties. Acid denaturation also showed similar structural changes in both forms of the enzyme. However, on alkaline treatment contrasting effects on the structure and stability of the two forms of enzyme were observed. The glycosylated enzyme undergoes partial unfolding with decreased stability at alkaline pH; however, a compaction of native conformation and enhanced stability of enzyme was observed for the deglycosylated enzyme under similar conditions. This is the first experimental demonstration of the influence of carbohydrate moiety on structure and stability of glucose oxidase. The studies also indicate the importance of pH studies in evaluating the effect of carbohydrate moiety on the structural and stability properties of glycoprotein. PMID- 12729621 TI - A role for the interleukin-1 receptor in the pathway linking static mechanical compression to decreased proteoglycan synthesis in surface articular cartilage. AB - Loading of articular cartilage during weight bearing is essential for the maintenance of cartilage function. Although certain cyclic loading protocols stimulate extracellular matrix synthesis, constant or static compression decreases proteoglycan and collagen synthesis in cartilage explants. The goal of this study was to determine whether the compression-induced decrease in proteoglycan synthesis involves an interleukin-1 (IL-1) signaling pathway. Cartilage explants were compressed 50% in the presence of IL-1 receptor antagonist (IL-1ra), and the incorporation of [35S]sulfate into macromolecules was measured. IL-1ra increased sulfate incorporation in compressed cartilage but not in cartilage maintained at the in situ thickness (0% compression). IL-1alpha and IL-1beta mRNAs were detected in cartilage compressed 50% for at least 3h, while nitric oxide synthase II mRNA was only detected in cartilage compressed 50% for 6h. The data support a role for the IL-1 receptor in the pathway linking static compression to reduced proteoglycan synthesis. PMID- 12729622 TI - Mutation of active site residues of the puromycin-sensitive aminopeptidase: conversion of the enzyme into a catalytically inactive binding protein. AB - The active site glutamate, Glu 309, of the puromycin-sensitive aminopeptidase was mutated to glutamine, alanine, and valine. These mutants were characterized with amino acid beta-naphthylamides as substrates and dynorphin A(1-9) as an alternate substrate inhibitor. Conversion of glutamate 309 to glutamine resulted in a 5000- to 15,000-fold reduction in catalytic activity. Conversion of this residue to alanine caused a 25,000- to 100,000-fold decrease in activity, while the glutamate to valine mutation was the most dramatic, reducing catalytic activity 300,000- to 500,000-fold. In contrast to the dramatic effect on catalysis, all three mutations produced relatively small (1.5- to 4-fold) effects on substrate binding affinity. Mutation of a conserved tyrosine, Y394, to phenylalanine resulted in a 1000-fold decrease in k(cat), with little effect on binding. Direct binding of a physiological peptide, dynorphin A(1-9), to the E309V mutant was demonstrated by gel filtration chromatography. Taken together, these data provide a quantitative assessment of the effect of mutating the catalytic glutamate, show that mutation of this residue converts the enzyme into an inactive binding protein, and constitute evidence that this residue acts a general acid/base catalyst. The effect of mutating tyrosine 394 is consistent with involvement of this residue in transition state stabilization. PMID- 12729623 TI - Purification and characterization of yeast mitochondrial initiation factor 2. AB - Yeast mitochondrial initiation factor 2 (ymIF2) is encoded by the nuclear IFM1 gene. A His-tagged version of ymIF2, lacking its predicted mitochondrial presequence, was expressed in Escherichia coli and purified. Purified ymIF2 bound both E. coli fMet-tRNA(f)(Met) and Met-tRNA(f)(Met), but binding of formylated initiator tRNA was about four times higher than that of the unformylated species under the same conditions. In addition, the isolated ymIF2 was compared to E. coli IF2 in four other assays commonly used to characterize this initiation factor. Formylated and nonformylated Met-tRNA(f)(Met) were bound to E. coli 30S ribosomal subunits in the presence of ymIF2, GTP, and a short synthetic mRNA. The GTPase activity of ymIF2 was found to be dependent on the presence of E. coli ribosomes. The ymIF2 protected fMet-tRNA(f)(Met) to about the same extent as E. coli IF2 against nonenzymatic deaminoacylation. In contrast to E. coli IF2, the complex formed between ymIF2 and fMet-tRNA(f)(Met) was not stable enough to be analyzed in a gel shift assay. In similarity to other IF2 species isolated from bacteria or bovine mitochondria, the N-terminal domain could be eliminated without loss of initiator tRNA binding activity. PMID- 12729624 TI - Heterologous expression of Ascaris suum cytochrome b5 precursor protein: a histidine-tagged full-length presequence is correctly processed to transport the mature protein to the periplasm of Escherichia coli. AB - The cytochrome b(5) of the body wall of adult Ascaris suum, a porcine parasitic nematode, is a novel type of cytochrome b(5). It is a soluble protein that lacks the COOH-terminal membrane-anchoring domain found in erythrocyte cytochrome b(5), but possesses an NH(2)-terminal extension (presequence) of 30 amino acids that are missing from the 82-residue protein purified from the nematode tissues [Yu, Y., Yamasaki, H., Kita, K., and Takamiya, S., 1996, Arch. Biochem. Biophys. 328, 165-172]. The nematode cytochrome b(5) is, therefore, probably synthesized as a precursor protein whose presequence is cleaved to form a mature protein, but the localization of the mature protein is still unknown. To investigate the processing of the putative precursor protein, the wild-type precursor of nematode cytochrome b(5) with a complete presequence (b5wt) and its NH(2) terminus truncated derivatives, b5Delta18 and b5Delta28, with 18 and 28 residues deleted, respectively, were expressed using pET-28a(+) vector in Escherichia coli. As expected, all transformants, tb5wt, tb5Delta18, and tb5Delta28, produced recombinant proteins with a histidine-tagged NH(2)-terminal extension. However, only the recombinant protein with the full-length presequence, produced in tb5wt, was correctly processed and transported to the periplasm, from which the majority of the induced product was purified as a mature protein chemically and functionally identical to the native protein purified from the nematode body wall. These results clearly show that the nematode histidine-tagged presequence functions as a signal peptide in E. coli. PMID- 12729625 TI - Taxoid metabolism: Taxoid 14beta-hydroxylase is a cytochrome P450-dependent monooxygenase. AB - The production of the anticancer drug Taxol in Taxus (yew) cell cultures is often accompanied by the formation of side-route polyoxygenated taxoid metabolites bearing a 14beta-hydroxyl group. The recent acquisition of several new semisynthetic taxoid intermediates enabled the screening of a family of Taxus cytochrome P450 cDNA clones for the 14beta-hydroxylase and additional taxoid oxygenases. The candidate cytochrome P450 clones were functionally expressed in yeast and tested by in vivo feeding of radiolabeled 5alpha-acetoxy-10beta-hydroxy taxadiene and 5alpha,13alpha-dihydroxy taxadiene. One clone efficiently and specifically transformed the 5alpha-acetoxy-10beta-ol, but not the 5alpha,13alpha diol, to a more polar product with the chromatographic properties of a taxoid triol monoacetate, and the identity of this product was confirmed by spectroscopic means as 5alpha-acetoxy-10beta,14beta-dihydroxy taxadiene. Microsome preparation from the transformed yeast allowed characterization of this new hydroxylase, which was shown to resemble other cytochrome P450 taxoid hydroxylases with pH optimum at 7.5 and a K(m) value for the taxoid substrate of about 50 microM. Because Taxol is unsubstituted at C14, the 14beta-hydroxylase cannot reside on the pathway to the target drug but rather appears to be responsible for diversion of the pathway to 14-hydroxy taxoids that are prominent metabolites of Taxus cell cultures. Manipulation of this hydroxylase gene could permit redirection of the pathway to increase flux toward Taxol and could allow the preparation of 13alpha,14beta-hydroxy taxoids as new therapeutic agents. PMID- 12729627 TI - Structure-activity relationship on human serum paraoxonase (PON1) using substrate analogues and inhibitors. AB - Substrate analogues based on the parent compounds paraoxon and phenyl acetate were tested on human serum paraoxonase (PON1) to explore the active site of the enzyme. Replacement of the nitro group of paraoxon with an amine or hydrogen, as well as electronic changes to the parent compound, converted these analogues into inhibitors. Introduction of either electron-withdrawing or donating groups onto phenyl acetate resulted in reduction in their rate of hydrolysis by PON1. PMID- 12729628 TI - The design of 8,8-dimethyl[1,6]naphthyridines as potential anticonvulsant agents. AB - Starting from a series of 7-linked tetrahydroisoquinoline derivatives, as exemplified by SB-270664, a new series of 8,8-dimethylnaphthyridine compounds has been identified. SAR studies around these attractive leads have provided compounds such as 12 which display excellent anticonvulsant activity and an encouraging pharmacokinetic profile in vivo. PMID- 12729629 TI - Novel synthesis of 2'-O-methylguanosine. AB - An efficient and chemoselective synthesis of 2'-O-methylguanosine (6) has been accomplished in high yield without protection of the guanine base. The salient feature of the synthesis of 6 lies in the application of methylene-bis (diisopropylsilyl chloride), (MDPSCl(2), 2) as a new 3',5'-O-protecting group for nucleosides. Use of CH(3)Cl as a weak electrophile and NaHMDS as a mild base was crucial to the success of the 2'-O-methylation of 3',5'-O-protected guanosine. PMID- 12729630 TI - Antibacterial activity of quinolone-macrocycle conjugates. AB - Novel quinolone-macrocycle conjugates displayed submicromolar antibacterial activity against Escherichia coli and Staphylococcus aureus bacterial strains. An analogous open-chain structure was not active at 100 microM against the same pathogenic strains. PMID- 12729632 TI - Study on the mechanism of action of artemether against schistosomes: the identification of cysteine adducts of both carbon-centred free radicals derived from artemether. AB - Reaction of the antimalarial and anti-schistosome drug artemether (1) and catalytic amount of ferrous ion in the presence of excess cysteine gave two adducts of cysteine and previous postulated primary and secondary carbon-centred free radicals besides their other rearrangement products. This piece of further evidence for the presence of carbon-centred radicals, especially the secondary carbon-centred free radical for the first time by the isolation of its coupling adduct, will be help to understand the mechanism of action of artemether and other qinghaosu derivatives against parasites. PMID- 12729631 TI - Conformationally restricted 3,4-diarylfuranones (2,3a,4,5 tetrahydronaphthofuranones) as selective cyclooxygenase-2 inhibitors. AB - A number of naphthofuranones were synthesized and tested for COX-1 and COX-2 inhibition. Few of them were identified as selective COX-2 inhibitors. Structure activity relationship studies within the series are discussed. PMID- 12729633 TI - Inactivation of S-adenosyl-L-homocysteine hydrolase with novel 5'-thioadenosine derivatives. Antiviral effects. AB - Synthesis of 5'-S-vinyl-5'-thioadenosine 5, 5'-S-ethynyl-5'-thioadenosine 7 and 5'-S-cyano-5'-thioadenosine 9 is described. Incubation of AdoHcy hydrolase with 5, 7 and 9 resulted in time- and concentration-dependent inactivation of the enzyme and partial depletion of its NAD(+) content. From these results and characterisation of metabolites released during the inactivation process, hypothetical mechanisms are suggested. The antiviral activity of 5, 7 and 9 was examined. Significant activities were noted with 5 against Vaccinia, Junin and Taccaribe viruses. PMID- 12729634 TI - A green fluorescent chemosensor for amino acids provides a versatile high throughput screening (HTS) assay for proteases. AB - The water soluble fluorescein-based ligand 1 forms a non-fluorescent complex with Cu(2+). This complex serves as a fluorescent sensor for amino acids in the 10(-3) M concentration range. Since the signal response is very fast, the sensor can be used to detect the hydrolytic activity of various proteases (trypsin, chymotrypsin, subtilisin) on bovine serum albumin as a whole protein substrate, and more generally to follow reactions releasing or removing free amino acids, in real time. PMID- 12729635 TI - Design and synthesis of spiro-cyclopentenyl and spiro-[1,3]-dithiolanyl substituted pyrrolidine-5,5-trans-lactams as inhibitors of hepatitis C virus NS3/4A protease. AB - Using the pyrrolidine-5,5-trans-lactam template, we have designed small, neutral, mechanism-based inhibitors of hepatitis C NS3/4A protease. Compound 2b, with a spiro-cyclobutyl P1 substituent and an isopropyl carbonyl substituent at the lactam nitrogen, has an IC(50) value in the replicon cell-based assay of 3 microM. PMID- 12729636 TI - Identification of a novel antiangiogenic agent; 4-(N-imidazol-2-ylmethyl)amino benzopyran analogues. AB - A series of 4-(N-imidazol-2-ylmethyl)aminobenzopyran analogues, originally designed as K(ATP) openers for ischemic diseases, showed antiangiogenic properties through the inhibition of HUVEC tube formation. Especially one of p-Cl substituted analogues (4c) completely inhibited HUVEC tube formation at 10 microM. The compound 4c significantly inhibited tumor growth by 52% on A549 (human non small cell lung carcinoma) in nude mice xenografts without any significant side effects. PMID- 12729638 TI - Identification of 2-aminobenzimidazole dimers as antibacterial agents. AB - The preparation and evaluation of 2-aminobenzimidazole dimers as antibacterial agents is described. Biological screening of the dimers indicated that compounds with multiple chloro substituents possessed optimal antibacterial activity. PMID- 12729637 TI - The development of new triazole based inhibitors of tumor necrosis factor-alpha (TNF-alpha) production. AB - 4-Aryl-5-pyridyl and 4-aryl-5-pyrimidyl based inhibitors of TNF-alpha production, which contain a novel triazole 5-member heterocyclic core, are described. Many pyridyl triazoles containing either an alkyl ether or a substituted aryl side chain on the triazole core showed sub-micromolar activity against LPS-induced TNF alpha, while pyrimidyl triazoles containing an ethoxymethyl side chain exhibited even better inhibitory activity. Secondary screening data are presented for the pyrimidyl triazoles. Triazole 14e combined excellent potency with good oral bioavailability in the rat. PMID- 12729639 TI - 2,4-disubstituted pyrimidines: a novel class of KDR kinase inhibitors. AB - 2,4-Disubstituted pyrimidines were synthesized as a novel class of KDR kinase inhibitors. Evaluation of the SAR of the screening lead compound 1 (KDR IC(50)=105 nM, Cell IC(50)=8% inhibition at 500 nM) led to the potent 3,5 dimethylaniline derivative 2d (KDR IC(50)=6 nM, cell IC(50)=19 nM). PMID- 12729640 TI - Novel 6,7-diphenyl-2,3,8,8a-tetrahydro-1H-indolizin-5-one analogues as cytotoxic agents. AB - A series of 6,7-diphenyl-2,3,8,8a-tetrahydro-1H-indolizin-5-one analogues were synthesized and evaluated for cytotoxic activity against eight human cancer cell lines. Compounds 18, 21, 28, 29, 30 and 31 showed cytotoxic activity with GI(50) values in the range of 2.1-8.1 microM concentration. Among these, compounds 21 and 28 exhibited good pharmacokinetic properties. These compounds were further evaluated for their in vivo efficacy in modified hollow fibre assay (HFA). PMID- 12729641 TI - Identification of potent and broad-spectrum antibiotics from SAR studies of a synthetic vancomycin analogue. AB - Dimeric vancomycin analogues based on a lead compound identified from a library of synthetic analogues of vancomycin have up to 60-fold greater activity than vancomycin against vancomycin-resistant Enterococcus faecium (VRE, VanA phenotype). Simplified analogues have also been prepared and found to maintain activity against VRE and have broad-spectrum antibiotic activity. PMID- 12729643 TI - Discovery of novel low molecular weight inhibitors of IMPDH via virtual needle screening. AB - Novel, low molecular weight inhibitors of IMPDH have been discovered through the application of a validated virtual screening protocol. A series of 21 IMPDH inhibitors were used to validate the docking procedure. Application of this procedure to the selection of compounds for screening from an in-house database resulted in a 50-fold reduction in the size of the screening set (3425 to 74 compounds) and gave a hit-rate of 10% on biological evaluation. PMID- 12729642 TI - Synthesis of beta-lactamase activated nitric oxide donors. AB - In order to achieve site specific delivery of NO, we designed conjugates of cephalosporin with NO donors. NO donors such as cupferron and SIN-1 were evaluated as potential choices for conjugates. Cephalosporin conjugated with SIN 1 demonstrated promising beta-lactamase dependent NO releasing ability. PMID- 12729644 TI - Synthesis of water-soluble prodrugs of BMS-191011: a maxi-K channel opener targeted for post-stroke neuroprotection. AB - A variety of water-soluble prodrugs of BMS-191011 was synthesized and evaluated for solution state stability and rate of conversion to BMS-191011 in rat and human plasma. The deoxycarnitine ester prodrug (11c) was selected for clinical evaluation based on its superior chemical stability, crystallinity and cleavage to BMS-191011 in human plasma. PMID- 12729645 TI - Spiro[pyrrolidine-2,2'-adamantanes]: synthesis, anti-influenza virus activity and conformational properties. AB - Synthetic spiro[pyrrolidine-2,2'-adamantanes] 2, 3, 11, 15, 12, 16, 18, 20 were evaluated in vitro and found to be active anti-influenza virus A compounds; the effect of the position of C-Me pyrrolidine ring substituent on antiviral activity was examined. Pyrrolidine 5-Me substitution appears to be optimal for H(2)N(2) strain activity. From the four different possible protonated conformers, experimental observation using NMR spectroscopy and molecular mechanics calculations demonstrated only a pair of conformers A(+)H (N-Me (ps-ax), C-Me (ps eq)) and B(+)H ((N-Me ps-ax, C-Me ps-ax)) which can contribute to the biological activity of C-Me, N-Me protonated derivatives 15(+)H, 16(+)H and 20(+)H. The relative populations were calculated from NMR spectra. For compounds 15(+)H and 20(+)H conformer A(+)H (cis dimethyl orientation) is the major one whereas a similar population of conformers A(+)H and B(+)H (trans dimethyl orientation) was observed for compound 16(+)H. Since this new series is characterized by a lipophilic part, that is the pyrrolidine ring, in addition to adamantane, that can interact with influenza A M2 protein, an ultimate future goal would be the in vitro mapping of M2 lipophilic pocket. PMID- 12729646 TI - New fibrinolytic agents: benzothiophene derivatives as inhibitors of the t-PA-PAI 1 complex formation. AB - The synthesis and activity of novel benzothiophene derivatives are described. In the t-Pa-induced fibrin clot lysis assay, several compounds inhibit the formation of the tPa-PAI-1 complex with submicromolar IC(50). This class of compounds potentially represents a new generation of antithrombotic-fibrinolytic agents. PMID- 12729647 TI - P-selectin blocking potency of multimeric tyrosine sulfates in vitro and in vivo. AB - P-selectin blocking potency was investigated using synthetic monomeric and polymeric anionic compounds containing sulfate groups such as O-sulfotyrosine (sTyr) and/or sulfated Lewis structures. A non-carbohydrate-containing polyacrylamide conjugate sTyr-PAA (80% mol of sTyr) was a remarkably potent inhibitor of P-selectin binding in vitro, having an IC(50) value of 6 ng/mL (equivalent to 10 nM calculated on the basis of sTyr residues or 0.1 nM calculated by the mass of the macromolecule). The inhibitory effect of sTyr-PAA (80%) towards P-selectin is significantly greater than that of fucoidan (IC(50), 100 ng/mL). However, sTyr-PAA (80%) was less effective than fucoidan at reducing neutrophil extravasation in an in vivo rat model of peritonitis. PMID- 12729648 TI - Design, synthesis and biological evaluation of hetaryl-nucleoside derivatives as inhibitors of chitin synthase. AB - We report here the design, synthesis and biological evaluation of new models of sugar analogues for chitin synthase. These UDP-GlcNAc mimetics associate a sugar mimicking hetaryl group and uridine, linked with different pyrophosphate bioisosteres. The compounds displayed weak inhibition activity on chitin synthase and their antifungal potencies have been assayed against a large variety of pathogenic fungi. PMID- 12729649 TI - Synthesis and pharmacological activity of fluorescent histamine H2 receptor antagonists related to potentidine. AB - Fluorescently labeled histamine H(2) receptor antagonists were synthesized starting from N-cyano-N'-[3-(3-piperidin-1-ylmethylphenoxy)propyl]guanidines with an additional N"-omega-aminoalkyl substituent (chain lengths 2-8 methylene groups) or from 3-(3-piperidin-1-ylmethylphenoxy)propylamine. The primary amino group was derivatized with various fluorophores (fluorescein, acridine, dansyl, nitrobenzoxadiazole (NBD), indolo[2,3-a]quinolizine). On the isolated spontaneously beating guinea pig right atrium most of the fluorescent probes were only weakly active, however, the NBD-labeled substances proved to be potent histamine H(2) receptor antagonists achieving pA(2) values in the range of 7.5 8.0, comparable to the activity of famotidine. PMID- 12729650 TI - Solid-phase synthesis of endothelin receptor antagonists. AB - A new solid-phase synthesis for ET receptor antagonists suitable for automation is presented. A support bound 2-hydroxybutyric acid derivative was converted to the corresponding ether derivatives using 4-halo-2-methylsulfonylpyrimidines. Subsequent Suzuki coupling with various aryl boronic acids gave the desired antagonists in good yields and purities. Highly potent antagonists with excellent selectivity for ET(A) were obtained. PMID- 12729651 TI - Synthesis of fluorinated 1-(3-morpholin-4-yl-phenyl)-ethylamines. AB - The synthesis of four (+/-)-fluorinated 1-(3-morpholin-4-yl-phenyl)-ethylamines and an enantioselective approach to these amines through reductive amination are described. PMID- 12729652 TI - Unique spirocyclopiperazinium salt. Part 2: synthesis and structure-activity relationship of dispirocyclopiperazinium salts as analgesics. AB - Three series of spirocyclopiperazinium derivatives 5a-d, 6a-f and 17a-d were synthesized and evaluated for their in vivo analgesic activities. Compounds 5a, 17a and 17b exhibited excellent analgesic activity. Two important structure activity relationships were observed from this study: (1) the quaternary ammonium functionality is a critical pharmacophore for analgesic activity; (2) it is important to adjust the lipophilic property of compounds to improve analgesic activity. PMID- 12729653 TI - EDTA bis-(methyl tyrosinate): a chelating peptoid peroxynitrite scavenger. AB - Conjugation of ethylenediaminetetra-acetic acid (EDTA) to methyl tyrosinate generates a chelating peptoid EDTA bis-(methyl tyrosinate), (EBMT). Peroxynitrite mediated nitration was studied for the free peptoid and its ferric and cupric complexes. The nitration products were monitored by electronic absorption spectroscopy at lambda(max) of 420 nm (mono-nitrated) and 440 nm (di-nitrated). Peak deconvolution was effected by pH manipulation as the mono-nitrated analogue of tyrosine exhibited a bathochromic shift from 365 nm (below its pK(a) of 6.8) to 420 nm. Rates of nitration were: free peptoid 10 mg/l) and ammonia (>0.01 mg unionized NH(3)/l), concentrations from organically enriched untreated wastewater discharges. However, taxa richness and abundance of macroinvertebrates were influenced greatly by the discharges. Where BOD and ammonia concentrations were elevated, the dominant taxa were oligochaete worms and chironominds. Fish and crustaceans (freshwater crabs and prawns) were found only in sites with the least BOD concentrations (furthest upstream and downstream). The maximum concentration of LAS (0.122 mg/l) was less than that expected to affect 5% of taxa (0.245 mg/l), whereas exceedences of DO and ammonia criteria were observed in several sites. The lack of recovery observed was attributed to influences of low DO, high ammonia and poor colonization from upstream and downstream reaches due to organically-enriched discharges PMID- 12729687 TI - Risk assessment approach for untreated wastewater using the QUAL2E water quality model. AB - This paper presents a novel approach for assessing the risk of consumer product ingredients in surface waters that receive untreated wastewater. The approach utilizes the water quality simulation model QUAL2E for predicting the impact caused by conventional pollutants, as well as the exposure concentration of consumer product ingredients. This approach invokes an impact zone concept whereby the receiving water can be thought of as a natural wastewater treatment system. After the river has recovered via self-purification, the ecosystem is then assessed by traditional risk assessment methods. This approach was validated using data collected from the Balatuin River, which is located in the Philippines. Results from this study support the use of QUAL2E for assessing the risk of consumer product ingredients in riverine systems receiving untreated wastewater. PMID- 12729688 TI - Deriving freshwater quality criteria for 2,4,6-trichlorophenol for protection of aquatic life in China. AB - Freshwater quality criteria of 2,4,6-trichlorophenol (2,4,6-TCP) were developed with particular reference to the aquatic biota in China, and based on USEPA's guidelines. Acute toxicity tests were performed on nine different domestic species indigenous to China to determine 48 h LC(50) and 96 h LC(50) values for 2,4,6-TCP. In addition, 21 d survival-reproduction test with Daphnia magna, 30 d embryo-larval test with Carassius auratus, 60 d fry-juvenile test with Ctenopharyngodon idellus, 30 d early life stage test with Bufo bufo gargarizans and 96 h growth inhibition test with Scenedesmus obliqaus were also conducted to estimate lower chronic limit and upper chronic limit values. The final acute value (FAV) was 2.01 mg/l 2,4,6-TCP. Acute-to-chronic ratios ranged from 5.01 to 12.2. The final chronic value (FCV) and the final plant value (FPV) of 2,4,6-TCP were 0.226 and 2.24 mg/l respectively. Based on FAV, FCV and FPV for 2,4,6-TCP, a criteria maximum concentration of 1.01 mg/l and a criterion continuous concentration of 0.226 mg/l were derived. The results of this study provide useful data for deriving national or local water quality criteria for 2,4,6-TCP based on aquatic biota in China. PMID- 12729689 TI - Influence of water hardening of the chorion on cadmium accumulation in medaka (Oryzias latipes) eggs. AB - This report describes a study in which in vitro fertilization methods were used to expose medaka (Oryzias latipes) eggs to cadmium (Cd(2+)). This approach was applied to address the differential sensitivity and cumulative potential of Cd(2+) when exposure was initiated early (before fertilization and water hardening of the chorion) versus later during embryo development (i.e., well after the chorion has undergone water hardening). Following range finding exposures (2.5, 10, 20, 40 or 80 mg/l) under artificially controlled experimental procedures, results from hatching success and embryo malformations showed the earlier exposure interval more sensitive than the assay involving only the embryonated egg. Subsequent accumulation studies have shown that the exposure initiated before fertilization apparently led to more Cd(2+) deposition in the chorion compared to the exposure during embryonated stages of the eggs. Similarly, values for total Cd(2+) indicated higher concentrations in those eggs exposed prior to--and during--water hardening. Results suggest an alteration of the properties of the zona radiata in the early-stage eggs, making it more permeable to the potential exit or entrance of waterborne agents even after water hardening. Ongoing studies must now address the development of more realistic exposure conditions of the gametes by using incubation media with osmolarities similar to surface waters, and by shortening duration for gamete exposure. Also, sensitive methods to localize Cd(2+) and to delineate the transfer from the chorion to the embryo are needed. PMID- 12729690 TI - Toxicity identification evaluation of leachates from municipal solid waste landfills: a multispecies approach. AB - The toxicity of leachates from two municipal solid waste (MSW) landfills in Southern Italy was characterized using a toxicity identification evaluation procedure. The chemical and physical fractionation techniques were: pH adjustment, pH adjustment/filtration, pH adjustment/C(18) solid phase extraction, graduated pH and EDTA chelation. All the samples exhibited acute toxicity towards the bacterium Vibrio fischeri, the freshwater rotifer Brachionus calyciflorus and the freshwater crustaceans Thamnocephalus platyurus and Daphnia magna. Statistical techniques were used to determine the discriminatory power and the toxicity detection capacity of the different assays and to choose a minimal battery of tests for the toxicity identification of leachates. Toxicity was closely associated with pH, generally increasing at higher pH levels and decliming at lower ones. Furthermore, results showed that toxicants could be characterized as cations, basic chemicals, suspended solids and apolar compounds. PMID- 12729691 TI - Cyst-based toxicity tests XVI--sensitivity comparison of the solid phase Heterocypris incongruens microbiotest with the Hyalella azteca and Chironomus riparius contact assays on freshwater sediments from Peninsula Harbour (Ontario, Canada). AB - In view of the complexity and costs of "traditional" whole sediment assays, a "culture/maintenance free" direct contact microbiotest has been developed with the freshwater ostracod Heterocypris incongruens. The new Toxkit assay (named Ostracodtoxkit) has been applied to 33 sediment samples from Peninsula Harbour, located in Lake Superior of the Great Lakes water basin in Ontario, Canada. The microbiotest was applied in parallel to direct contact tests with the amphipod Hyalella azteca and the midge larva Chironomus riparius, to compare its relative sensitivity with that of the two "conventional" assays. The study was undertaken in the framework of remediation action plans for specific areas of concern, to enable decision making by the Canadian authorities for the restoration of impacted aquatic environments. Most sediments were found non-toxic (<20% mortality) to both the conventional test species and the ostracod. For the large majority of samples, a very good correspondence was found between the two crustacean test species for the intensity of the toxic signal "mortality", as reflected by a 0.71 (p<0.05) correlation coefficient. Growth inhibition, which is determined in the ostracod microbiotest as a sublethal effect criterion, allowed the earmarking of some sediment samples, which were apparently more toxic for the amphipod than to the ostracod. For 20% of the samples, substantially higher mortality scores were noted with the ostracod assay than with the midge larvae tests and the overall correlation coefficient between these two tests was lower (r=0.60,p<0.05). The results obtained in the present study corroborate those of previous research on sediments collected from various rivers in Flanders, Belgium, and confirm the potential of the new ostracod microbiotest as a reliable and sensitive low cost alternative for traditional whole sediment assays. PMID- 12729692 TI - Specific detection of organotin compounds with a recombinant luminescent bacteria. AB - Organotin compounds are widely used as biocides in marine and terrestrial environments. Several currently used techniques allow either the measurement of the chemicals or their effects on living organisms. Our current research focuses on the development of a complementary method based on a bacterial bioluminescence based bioassay for the specific detection of organotin compounds. The performance of the bioassay was assessed. The Escherichia coli bacterial strain used in this study is specific for TBT and DBT (with Cl, Br or I as the halogen group) with the central tin atom important for light production. The assay is conducted after overnight culture of the bacterial strain, followed by 60 min of contact time with the organotin compound for significant light production. The detection limits were found to be 0.08 microM for TBT (26 microgl(-1)) and 0.0001 microM for DBT (0.03 microgl(-1)) with a linear range of one logarithm. The repeatability of the bioassay is 8% and the reproducibility for TBT and DBT was approximately 14%. Lyophilization of the strains did not significantly modify the detection limit as well as the range of detection. Applications of the bioassay to environmental samples are discussed. PMID- 12729693 TI - Lanthanum fixation by Myxococcus xanthus: cellular location and extracellular polysaccharide observation. AB - Myxococcus xanthus is a soil bacterium of the myxobacteria group and is abundant in almost all soils. Its role in soil ecology is considered significant. One noteworthy characteristic of the bacterium is that it produces large quantities of extracellular polymeric substances (EPS). It is also known that its biomass has the capacity to fix heavy metals. Here it is reported that M. xanthus was able to accumulate 0.6 mmol of La per g of wet biomass and/or 0.99 mmol per g of dry biomass. Transmission Electron Microscopy (TEM) observation of M. xanthus cells treated with La showed that a substantial amount of this cation was fixed in the EPS and in the cell wall. Smaller amounts were also observed in the cytoplasm. Fixed La appeared as phosphate in all cellular locations. The results given here also show that the use of La enables TEM observation of the M. xanthus EPS as a dense fibrillar net surrounding the cells. This technique is relatively easy and prevents EPS collapse, which occurs frequently during the fixation and dehydration procedures commonly used in preparations for TEM observations. Since antibodies are no longer required, the La stain can be carried out without delaying bacterial cell cultivation or isolation. In addition, the presence of La in cell cytoplasm without cell degeneration suggests that this microorganism could be used as a model in the study of bacteria-lanthanide interactions. PMID- 12729694 TI - In situ monitoring of urban air in Southern Italy with the Tradescantia micronucleus bioassay and semipermeable membrane devices (SPMDs). AB - This study was designed to assess toxic and genotoxic compounds in the urban air of Caserta, South Italy using cuttings from the plant Tradescantia #4430. In situ monitoring of gaseous pollutans was made at 17 sampling points in two seasons of the year. Genotoxicity was evaluated by recording the micronuclei in meiotic pollen mother cells (Trad-MCN assay). In addition, the passive sampler semipermeable membrane devices (SPMDs) were deployed at the sampling points with a significant increase in micronuclei frequency. SPMDs concentrated priority organic pollutants were identified by high performance liquid chromatography and gas chromatography, while toxicity and mutagenesis were assessed on the bacterium Vibrio fischeri using the Microtox and Mutatox systems respectively. Significant toxic and mutagenic effects were observed at different points on the town grid and SPMDs effectively concentrated trace contaminants. The relationship between what was present in the air sampled by SPMDs and the micronuclei frequency was also explored. PMID- 12729695 TI - Evaluation of the effect on arthropod populations by using deltamethrin to control Phloeotribus scarabaeoides Bern. (Coleoptera: Scolytidae) in olive orchards. AB - Field tests were made for the control of an olive insect pest, the olive bark beetle, Phloeotribus scarabaeoides (Col: Scolytidae), using the pyrethroid insecticide deltamethrin (D) and this insecticide combined with ethylene (D+E), an attractant of the olive bark beetle. The tests were run in olive orchards in the provinces of Granada and Jaen (Andalusia, southern Spain), treating several olive rows so as to create a barrier effect to control the pest attack. The main objective was to evaluate the arthropod fauna affected by these treatments. The different taxa captured have been separated in three different trophic groups: parasitoids, predators and phytophagous. A knock-down effect was found during the first dates in all the functional groups in all cases. The results showed the following trend in arthropod abundance: D+E>D>C. This trend was significant (p<0.05) for predators and phytophagous insects in both zones. The parasitoids of Prays oleae (Lep.: Plutellidae), a major olive pest, were most affected by the treatments, together with several groups of predators such as ants, cantharids, coccinellids and mirids. Data from the captures show the need to determine the optimum time for insecticide application in order to minimise its effect on beneficial insect populations. PMID- 12729696 TI - Waterborne and sediment toxicity of fluoxetine to select organisms. AB - Ecological risk assessments of pharmaceuticals are currently difficult because little-to-no aquatic hazard and exposure information exists in the peer-reviewed literature for most therapeutics. Recently several studies have identified fluoxetine, a widely prescribed antidepressant, in municipal effluents. To evaluate the potential aquatic toxicity of fluoxetine, single species laboratory toxicity tests were performed to assess hazard to aquatic biota. Average LC(50) values for Ceriodaphnia dubia, Daphnia magna, and Pimephales promelas were 0.756 (234 microg/l), 2.65 (820 microg/l), and 2.28 microM (705 microg/l), respectively. Pseudokirchneriella subcapitata growth and C. dubia fecundity were decreased by 0.044 (14 microg/l) and 0.72 microM (223 microg/l) fluoxetine treatments, respectively. Oryias latipes survival was not affected by fluoxteine exposure up to a concentration of 28.9 microM (8.9 mg/l). An LC(50) of 15.2 mg/kg was estimated for Chironomus tentans. Hyalella azteca survival was not affected up to 43 mg/kg fluoxetine sediment exposure. Growth lowest observed effect concentrations for C. tentans and H. azteca were 1.3 and 5.6 mg/kg, respectively. Our findings indicate that lowest measured fluoxetine effect levels are an order of magnitude higher than highest reported municipal effluent concentrations. PMID- 12729697 TI - Evaluation and further development of the activated sludge respiration inhibition test. AB - The activated sludge respiration inhibition test is an important bacterial test system for the determination of bacterial toxicity of chemical compounds. The exposure period recommended by OECD 209 and ISO 8192 protocols is 30 and 180 min. A modified version of the test was developed which allowed a prolonged incubation period of 27 h to enhance the possibilities of the test system. The test system with the prolonged incubation time was evaluated by the recommended reference compound 3.5-dichlorophenol and showed an EC50 of 6.3 mgl(-1) with a coefficient of variation of 12.7%. Furthermore, the use of an open test system was evaluated showing a comparable toxicity but a higher coefficient of variation than the closed test system. A storage of activated sludge for several days accompanied by a daily feed with OECD nutrient solution should be avoided, for it can cause a decreased sensitivity of the inoculum. Different statistical fit procedures were tested indicating that Weibull fit procedures were superior for extended data sets covering a wide range of concentrations whereas Gamma and Probit fits were appropriate for smaller data sets mainly restricted to the linear part of the dose response curve. PMID- 12729698 TI - Degradation of juvenile hormone analog by soil microbial isolates. AB - Juvenoids are efficient pesticides with relatively low toxicity to humans. However, few studies have evaluated the effect of degradation by soil microorganisms on their toxicity. The effects of bacterial, fungal and yeast isolates on aerobic decomposition of ethyl N-[2-[4-(2,2-ethylenedioxy-1 cyclohexylmethyl)phenoxy]ethyl] carbamate during eight weeks were determined. The effect of different concentration of glucose on their degradation activity is also analyzed. PMID- 12729699 TI - In situ cadmium reclamation by freshwater bivalve Lamellidens marginalis from an industrial pollutant-fed river canal. AB - The biofilter potential of the freshwater bivalve, Lamellidens marginalis was examined in cage experiments conducted in a river canal (Ichhapore, 24-Parganas, West Bengal, India) receiving industrial effluents from steel and metal factories as well as from an ordinance factory. Cadmium is one of the major contaminants in this river canal. Lamellidens collected from pollution free natural ponds, were sorted into three size groups (large: 59+/-3.2 g, 10+/-2.3 cm; medium: 30+/-2 g, 6+/-1.7 cm and small: 13+/-1.5 g, 4+/-1.2 cm) were held in cages at three different sites along a cadmium concentration gradient. Concentrations of cadmium were measured from water, sediment and different tissues of Lamellidens at weekly intervals using atomic absorption spectrophotometric methods. Cadmium uptake by Lamellidens in all media were highly concentration dependent in both summer and winter months. For all three size groups, cadmium uptake was maximum in the gills at the beginning of experiment, and liver at the later phase. Cadmium uptake was maximum in the small bivalves and minimum in the large bivalves groups. Cadmium uptake was 11-67% higher during summer than during the monsoon season for all tissues and size groups. Estimation of concentration factor revealed that tissues were saturated with cadmium during the 13-14th week after Lamellidens introduction during summer, but remained unsaturated during the monsoon season. It is concluded that Lamellidens might be considered as an efficient biofilter for reclamation of aquatic environment having sub-lethal concentrations of cadmium. PMID- 12729700 TI - Microbiological reduction of chromium(VI) in presence of pyrolusite-coated sand by Shewanella alga Simidu ATCC 55627 in laboratory column experiments. AB - Hexavalent chromium (Cr(VI)) was reduced to non-toxic trivalent chromium (Cr(III)) by a dissimilatory metal reducing bacteria, Shewanella alga Simidu (BrY MT) ATCC 55627. A series of dynamic column experiments were conducted to provide an understanding of Cr(VI) reduction by the facultative anaerobe BrY-MT in the presence of pyrolusite (beta-MnO(2)) coated sand and uncoated-quartz sand. All dynamic column experiments were conducted under growth conditions using Cr(VI) as the terminal electron acceptor and lactate as the electron donor and energy source. Reduction of Cr(VI) was rapid (within 8 h) in columns packed with uncoated quartz sand and BrY-MT, whereas Cr(VI) reduction by BrY-MT was delayed (57 h) in the presence of beta-MnO(2)-coated sand. The role of beta-MnO(2) in this study was to provide oxidation of trivalent chromium (Cr(III)). BrY-MT attachment was higher on beta-MnO(2)-coated sand than on uncoated quartz sand at 10, 60, and 85.5 h. Results have shown that this particular strain of Shewanella did not appreciably reduce Mn(IV) to Mn(II) species nor biosorbed Cr and Mn during its metabolic activities. PMID- 12729701 TI - Separation of the strength and selectivity of the microbiological effect of synthetic dyes by spectral mapping technique. AB - The growth inhibitory effect of 30 synthetic dyes on 22 bacteria (test organisms) belonging to various taxonomic groups was determined. The strength (potency) and selectivity of the biological effect were separated by the spectral mapping technique, reducing the dimensionality of the selectivity maps to two by the nonlinear mapping technique. The relationship between biological effect and physicochemical parameters of dyes was elucidated by stepwise regression analysis. It has been established that the strength of the effect of anthracene and trityl derivatives was higher than that of azobenzene dyes and significantly depended on the hydrophobicity of the compound. The selectivity of the effect also depended on hydrophobicity and on the nonpolar unsaturated surface area of the dyes. Gram negative and Gram positive bacteria differed in the strength and selectivity of their response to dyes indicating the marked impact of the taxonomical position on the response. Contrary to other multivariate mathematical statistical methods biological activity may be divided by SPM into potency and selectivity values, therefore, application of the technique in future QSAR studies is highly recommended. PMID- 12729702 TI - Investigation of acute toxicity and the effect of 2,4-D (2,4 dichlorophenoxyacetic acid) herbicide on the behavior of the common carp (Cyprinus carpio L., 1758; Pisces, Cyprinidae). AB - A 96-h LC(50) values of 2,4-D [(2,4-dichlorophenoxy)acetic acid], a common contaminating agricultural herbicide, were determined on the adult common carp (Cyprinus carpio L., 1758; Pisces, Cyprinidae). The study was conducted in two stages using 130 carp. The data obtained were statistically evaluated by the use of the EPA computer program based on Finney's Probit Analysis Method and a 96-h LC(50) value for C. carpio L., 1758 was found to be 63.24 mg/l in a static bioassay test system. 95% lower and upper confidence limits for the LC(50) were 55.03 and 71.92 mg/l, respectively. Water temperature was 17+/-1 degrees C. Behavioral changes of the above mentioned species were examined for various herbicide concentrations. PMID- 12729703 TI - Use of oxytetracycline and tylosin in intensive calf farming: evaluation of transfer to manure and soil. AB - Antibiotics may enter soils with manure from treated animals. Because of their biological effects, antibiotics are regarded as potential micropollutants. The levels of oxytetracycline and tylosin over time were followed in faeces, bedding and manure, and then in the soil of a manured field and surrounding drainage courses, after oral treatment of calves. Fifty Simmental calves were treated for 5 days with 60 mg/kg/day of oxytetracycline. After 15 days the animals were treated for 5 days with 20 mg/kg/day of tylosin. Tylosin degraded rapidly, and was no longer detected in manure 45 days after cessation of treatment and no trace of the compound was detected in soil or surrounding water (detection limits 10 microg/l). The half-life of oxytetracycline in manure was 30 days and the compound was still detectable in this matrix (820 microg/kg) after 5 months maturation. In the manured soil oxytetracycline was detected at concentrations at least 10 times lower than the European Agency for the Evaluation of Medicinal Products threshold (100 microg/kg) requiring phase II environmental risk assessment. Oxytetracycline was not detected in the water courses (detection limit 1 microg/l). These results demonstrate that the processes occurring between faeces production and application of manure to the soil are very effective in reducing the load of TYL and OTC in the environment. For both drugs a toxicity test was performed using the alga Selenastrum capricornutum. The EC50 was 4.18 mg/l for oxytetracycline and 0.95 mg/l for tylosin. A worst-case hazard assessment for the aquatic environment was performed comparing the ratio between the measured concentrations (LOD) and effect data from previous work (OTC) or from this work (TYL). This showed ratio between toxicity levels (bacteria) (EC50=0.14 mg/l) and measured concentrations (LOD=1 microg/l) for OTC to be 140. The corresponding value for TYL (LOD=10 microg/l) was 95. PMID- 12729704 TI - Effects of dietary habits and CYP1A1 polymorphisms on blood dioxin concentrations in Japanese men. AB - The major source of dioxin impurities in Japan in the past was agrochemical formulations; more recently, it has been exhaust from waste incinerators. To examine the environmental and genetic factors that influence blood dioxin concentration, we investigated the relationship among dioxin concentrations, dietary habits and cytochorome P450 1A1 (CYP1A1) polymorphisms (MspI type and Ile Val type) in Japanese fishermen and farmers, including also a group of office workers as controls. The mean dioxin concentrations in the fishermen, the farmers and the controls were 161369, 79079 and 100500 pg/g fat, respectively. The elevated dioxin concentration with polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans and coplanar-PCBs found in the fishermen may be due to the frequent consumption of fish; no such relationship was found both in the farmers and the controls. We found that the concentrations of congeners found as impurities in certain chemicals such as those previously used in agriculture showed no significant differences among the three groups; we consider it unlikely that the farmers would be directly exposed to dioxins from such chemicals. Thus, it is probable that the primary route of dioxin exposure in the Japanese population is through the food chain via fish consumption, regardless of occupation. No meaningful relationship between blood dioxin concentration and CYP1A1 polymorphisms was found in this study, although there was a significant difference between the concentration of total non-ortho-PCBs in genotypes A and B. Further studies on more subjects, including those of genotype C, are needed to confirm the relationship between blood dioxin concentrations and MspI polymorphisms. PMID- 12729705 TI - A new method to study biodegradation kinetics of organic trace pollutants by activated sludge. AB - A reliable prediction of the behaviour of organic trace compounds in activated sludge plants requires an accurate input of the biodegradation kinetics. Often these kinetics are extrapolated from the results of standardised biodegradation tests. However, these tests generally are not designed to yield kinetic information and do not reflect the conditions in activated sludge plants. To overcome these problems a new test method was developed which is referred to as a 'by-pass' test. The test methodology is explained and examples are given for three compounds: the C(12)-homologue of linear alkylbenzene sulfonate, nitrilotriacetic acid and toluene. More experience with the test is required, particularly with respect to selection of the proper test settings, which are compound related. The test is a suitable tool in a research environment, for example to investigate the effect of plant operational parameters on the biodegradation kinetics. PMID- 12729706 TI - Organochlorine pesticides, PCBs and heavy metals in tissues of the mullet Liza aurata in lake Ganzirri and Straits of Messina (Sicily, Italy). AB - The aim of this research was to investigate the presence of organochlorine pesticides, PCBs and heavy metals (Cd, Cr, Cu, Ni, Pb and Zn) in fish living in lake Ganzirri and the Straits of Messina, using the mullet Liza aurata as a "biological indicator". Different tissues of Liza aurata, which include the muscle, gill and vertebral column, were taken for analyses. Quantitative determination of the organochlorine compounds was performed by GC-ECD and confirmed with GC-MS. The concentrations of "essential" and "toxic" heavy metals were determined by Atomic Absorption Spectrophotometry. For the mullets of lake Ganzirri, the study showed DDE in three samples of gills and one of muscle, at concentrations below the MRL; no appreciable residues of PCBs (Aroclor 1232 series) were found. The concentrations of Zn and Cu were much higher than those of the "toxic" metals. No traces of organochlorine pesticides and PCBs were found in the individuals from the Straits of Messina. Therefore, these results indicate that these two environments are not at toxicological risk. PMID- 12729707 TI - Phenolic compounds content in Pinus halepensis Mill. needles: a bioindicator of air pollution. AB - Foliar phenol concentrations (total and simple phenols) were determined in Aleppo pine (Pinus halepensis Mill.) needles collected in June 2000, from 6 sites affected by various forms of atmospheric pollutants (NO, NO(2), NO(x), O(3) and SO(2)) monitored during two months. Results show an increase in total phenol content with exposure to sulphur dioxide and a reduction with exposure to nitrogen oxide pollution. p-Coumaric acid, syringic acid and 4-hydroxybenzoic acid concentrations increase with exposure to nitrogen oxide pollution, whereas gallic acid and vanillin decrease in the presence respectively of sulphur dioxide and ozone. This in situ work confirms the major interest of using total and simple phenolic compounds of P. halepensis as biological indicators of air quality. PMID- 12729708 TI - Influence of pH on the toxicity of nitrophenols to Microtox and Spirotox tests. AB - The toxicity of mononitrophenols and dinitrophenols (DNP) to luminescent bacteria Vibrio fischeri (Microtox test) and ciliated protozoan Spirostomum ambiguum (Spirotox test) was evaluated. Spirotox was more sensitive to the tested nitrophenols (NPs) than the Microtox test. 2,5-DNP was the most toxic and 2-NP was the least toxic to the both bioindicators. The toxicity depended greatly on the pH of the medium. The highest changes were observed for DNPs, where the toxicity decreased more than 20-times when the pH increased from 6 to 8. No significant decrease of the toxicity was found for NPs, when the pH increased from 6 to 7. Greater increase of the pH to 8 caused from 1.5 to 4-fold decrease of the toxicity. PMID- 12729710 TI - Investigation of acute toxicity of cadmium chloride (CdCl2 . H2O) metal salt and behavioral changes it causes on water frog (Rana ridibunda Pallas, 1771). AB - The 96-h LC(50) value of cadmium chloride (CdCl(2).H(2)O), a metal salt widely used in industry, was determined in the water frog (Rana ridibunda Pallas, 1771). The experiments were conducted in two series and a total of 140 frogs were used to determine acute toxicity. In addition, behavioral changes in the water frog were determined for each cadmium chloride concentration. Data obtained from the cadmium chloride investigation were evaluated by using the probit analysis statistical method and the 96 h LC(50) value for water frog was estimated to be 51.2 mg/l. PMID- 12729709 TI - Emission factors and sources of ethylene-diaminetetraacetic acid in waste water- a case study. AB - Ethylene-diaminetetraacetic acid (EDTA) is a complexing agent and has the ability to form stable water-soluble complexes with metal ions. It is used in a variety of industrial applications including pulp and paper, metal, textile, leather rubber, pharmaceuticals, food, polymer production and others. Most of these applications are water based and lead to emissions into the waste water and reach sewage treatment plants. Industrial sources and municipal waste waters were monitored simultaneously. Mixed samples were taken over periods of one week at nine sample sites. The concentrations of EDTA were measured in waste water samples by gas chromatography using N-selective detection. The results showed that, although, the concentrations and loads were variable the paper manufacturing industry was the major EDTA contributor to the influent of the waste water treatment plant and contributed more than 98% of the total load. All the other sources including two household areas, were comparably low. In waste water of households concentrations between 10 and 70 microg/l EDTA could be detected. Concentrations of EDTA from different industrial waste water sources ranged from 28 up to 3980 microg/l. Influent and effluent concentrations of the WWTP were usually high in the range of 500-940 and 390-760 microg/l; respectively. Elimination rates averaged 15%, the calculation is based on emission loads. Specific emission factors were calculated based on population equivalents. PMID- 12729711 TI - Potential availability of heavy metals to phytoextraction from contaminated soils induced by exogenous humic substances. AB - Effective phytoremediation of soils contaminated by heavy metals depends on their availability to plant uptake that, in turn, may be influenced by either the existing soil humus or an exogenous humic matter. We amended an organic and a mineral soil with an exogenous humic acid (HA) in order to enhance the soil organic carbon (SOC) content by 1% and 2%. The treated soils were further enriched with heavy metals (Cu, Pb, Cd, Zn, Ni) to a concentration of 0, 10, 20, and 40 microg/g for each metal and allowed to age at room temperature for 1 and 2 months. After each period, they were extracted for readily soluble and exchangeable (2.5% acetic acid), plant-available (DTPA, Diethylentriaminepentaacetic acid), and occluded (1 N HNO(3)) metal species. Addition of HA generally reduced the extractability of the soluble and exchangeable forms of metals. This effect was directly related to the amount of added HA and increased with ageing time. Conversely, the potentially plant available metals extracted with DTPA were generally larger with increasing additions of exogenous HA solutions. This was attributed to the formation of metal-humic complexes, which ensured a temporary bioavailability of metals and prevented their rapid transformation into insoluble species. Extractions with 1 N HNO(3) further indicated that the added metals were present in complexes with HA. The observed effects appeared to also depend on the amount of native SOC and its structural changes with ageing. The results suggest that soil amendments with exogenous humic matter may accelerate the phytoremediation of heavy metals from contaminated soil, while concomitantly prevent their environmental mobility. PMID- 12729712 TI - Bactericidal effect of TiO2 photocatalyst on selected food-borne pathogenic bacteria. AB - Titanium dioxide (TiO(2)) photocatalysts have attracted great attention as a material for photocatalytic sterilization in the food and environmental industry. This research aimed to design a new photobioreactor and its application to sterilize selected food borne pathogenic bacteria, Salmonella choleraesuis subsp., Vibrio parahaemolyticus, and Listeria monocytogenes. The photocatalytic reaction was carried out with various TiO(2) concentrations and Ultraviolet (UV) illumination time. A feasible synergistic effect was found that the bactericidal effect of TiO(2) on all bacterial suspension after UV light irradiation was much higher than that of without TiO(2). As the concentration of TiO(2) increased to 1.0 mg/ml, bactericidal effect increased. However, the bactericidal effect was rapidly abbreviated at TiO(2) concentration higher than 1.25 mg/ml to all selected bacteria. UV illumination time affected drastically the viability of all bacteria with different death rate. Similar trends were obtained from S. choleraesuis subsp. and V. parahaemolyticus that their complete killing was achieved after 3 h of illumination. However, L. monocytogenes was more resistant and its death ratio was about 87% at that time. PMID- 12729713 TI - Testicular histopathology in deer mice (Peromyscus maniculatus) following exposure to polychlorinated biphenyl. AB - Adult deer mice testes were subjected to routine histopathology following exposure to Aroclor 1254 supplemented diet (5 ppm), for 30 days. Body and testicular weight revealed no statistical significance between the control and treated animals. From a histological standpoint the testes of the controls were similar to normal murids and other animals. In contrast, the testes from treated animals displayed seminiferous tubules with significant degenerative alterations. These alterations included fewer layers of seminiferous epithelium exaggerated intercellular spaces and appearance of pyknotic nuclei. Most tubules displayed subluminal nuclei that morphologically could be identified as part of spermatozoa heads and these usually lacked tails, indicating that the treatment interfered with spermiogenesis. Therefore, we concluded that Aroclor 1254 as an environmental contaminant is highly destructive to seminiferous tubules, and that these histological alterations undoubtedly are responsible for the depressed fertility in Peromyscus following chronic exposure to PCBs, that has been reported in the literature. PMID- 12729714 TI - Acrylamide axonopathy revisited. AB - Distal swelling and eventual degeneration of axons in the CNS and PNS have been considered to be the characteristic neuropathological features of acrylamide (ACR) neuropathy. These axonopathic changes have been the basis for classifying ACR neuropathy as a central-peripheral distal axonopathy and, accordingly, research over the past 30 years has focused on the primacy of axon damage and on deciphering underlying mechanisms. However, based on accumulating evidence, we have hypothesized that nerve terminals, and not axons, are the primary site of ACR action and that compromise of corresponding function is responsible for the autonomic, sensory, and motor defects that accompany ACR intoxication (NeuroToxicology 23 (2002) 43). In this paper, we provide a review of data from a recently completed comprehensive, longitudinal silver stain study of brain and spinal cord from rats intoxicated with ACR at two different daily dosing rates, i.e., 50 mg/kg/day, ip or 21 mg/kg/day, po. Results show that, regardless of dose rate, ACR intoxication was associated with early, progressive nerve terminal degeneration in all CNS regions and with Purkinje cell injury in cerebellum. At the lower dose-rate, initial nerve terminal argyrophilia was followed by abundant retrograde axon degeneration in white matter tracts of spinal cord, brain stem, and cerebellum. The results support and extend our nerve terminal hypothesis and suggest that Purkinje cell damage also plays a role in ACR neurotoxicity. Substantial evidence now indicates that axon degeneration is a secondary effect and is, therefore, not pathophysiologically significant. These findings have important implications for future mechanistic research, classification schemes, and assessment of neurotoxicity risk. PMID- 12729715 TI - Pulmonary toxicity of polymeric hexamethylene diisocyanate aerosols in mice. AB - The acute pulmonary response of male C57BL/6 mice exposed to respirable polymeric hexamethylene diisocyanate biuret trimer aerosol (HDI-BT), a component of polyurethane spray paints, was examined. Mice were exposed to concentrations of 1 and 10 mg/m(3) HDI-BT for 5 h and were evaluated 6, 18, 42, 90, 186, and 378 h after the end of exposure. Mice exposed to 1 or 10 mg/m(3) HDI-BT exhibited dose dependent lung function impairment, edema, neutrophilic inflammation, cellular proliferation, and histologic lesions in terminal bronchioles and alveolar ducts. Impairment of pulmonary function, indicated by decreased frequency and increased enhanced pause (Penh), was maximal immediately after exposure and progressively recovered at later time points. Lung weight and lavage fluid protein content peaked at 6 and 18 h after exposure, respectively. Total cells and macrophages recovered in lavage fluid peaked 90 h after exposure. Neutrophils recovered in lavage fluid peaked between 18 and 42 h after exposure. Proliferative lesions, as identified histologically and by bromodeoxyuridine incorporation, were maximal 90 h after exposure. In contrast, no inflammatory cell influx, protein leakage, or lung pathology were observed in mice exposed to 360 ppb HDI monomer vapor. This model will be useful for investigating molecular mechanisms by which HDI-BT causes lung injury, which is known to occur in humans exposed occupationally to this pulmonary toxicant. PMID- 12729716 TI - Intestinal and hepatic lesions in mice, rats, and other laboratory animals after intravenous administration of gas-carrier contrast agents used in ultrasound imaging. AB - Single intravenous administration of three different gas-carrier contrast agents used in ultrasound imaging to mice caused inflammation, necrosis, and ulceration of cecum and proximal colon (cecocolonic area) and focal necrosis in the liver. Similar intestinal lesions were also found in rats after treatment with a single iv administration of a gas-carrier contrast agent. Strain differences in the incidences of these lesions were found in both rats and mice. HsdHan:NMRI mice were among the most sensitive of the strains of mice studied. Even at the lowest dose of Sonazoid technically possible to inject in HsdHan:NMRI mice, lesions were found and a no-effect dose could not be determined. In a time-course experiment in HsdHan:NMRI mice, it was found that the lesions began to develop in the cecum and colon within 15 to 30 min after dosing. Lesions in the liver were first observed 120-240 min after dosing. Diet played a role in the etiology of the lesions, as HsdHan:NMRI mice given a diet with reduced amounts of cellulose and starch had reduced incidences of lesions, and when glucose was the only carbohydrate source, no lesions were observed. No intestinal or hepatic lesions were found in guinea pigs or rabbits after repeated intravenous administrations of Sonazoid. In dogs, minimal to mild granulocytic inflammation of the cecum and/or colon was found after daily repeated intravenous injections for 28 days, but not after daily repeated administration for 14 days nor after a single administration. It is proposed that the intestinal and hepatic lesions in rats and mice after a single intravenous injection of gas-carrier contrast agents are caused by a common mechanism: intravascular growth of gas-carrier agents in tissues with gas supersaturation, as occurs in the cecal wall of rats and mice. In this particular environment the growing gas bubbles cause ischemia and necrosis in the cecal and colonic wall and liver. This proposed mechanism of action is consistent with the absence of clinical reports indicative of intestinal and/or hepatic lesions in humans after administration of gas-carrier contrast agents. PMID- 12729717 TI - Etiology of cecal and hepatic lesions in mice after administration of gas-carrier contrast agents used in ultrasound imaging. AB - The aim of the study was to investigate the etiology of cecal and hepatic lesions in mice and rats after intravenous administration of gas-carrier contrast agents (GCAs). A modified fluorescein flowmetry technique and 24 h necropsy were used in mice (conventional and germ free), rats, and guinea pigs after GCA administration. Different diets and oral nonabsorbable antibiotics were used. Nonfluorescence, edema, congestion, hemorrhage, and mucosal erosion in cecum and colon and nonfluorescent areas in the liver were observed from 16 min after GCA administration in conventional mice on standard diet. Numerous gas bubbles (>50 microm) were observed in the vasculature around the nonfluorescent areas of cecum and colon and in mesenteric vessels draining to the portal vein. Acute inflammation, edema, hemorrhage, and ulceration of the cecum and colon and liver necrosis were seen 24 h after GCA administration in conventional mice on standard diet. When mice were maintained on either a diet with glucose as the only carbohydrate source or on a standard diet supplemented with antibiotics, uniform fluorescence and no organ lesions were observed after GCA administration. Uniform fluorescence and no organ lesions were observed in germ-free mice, rats, and guinea pigs dosed with GCAs and in control animals (mice, rats, and guinea pigs) dosed with sucrose. The results indicate that intravascular growth of GCA microbubbles occurs in the cecal and colonic wall of mice, leading to occlusive ischemia and necrosis in these intestinal segments and secondary gas embolisation in the liver. Transmural gas supersaturation in the cecal wall may explain the intravascular bubble growth in mice. PMID- 12729718 TI - Comprehensive analysis of gene expression in rat and human hepatoma cells exposed to the peroxisome proliferator WY14,643. AB - Peroxisome proliferators (PPs) are an important class of chemicals that act as hepatic tumor promoters in laboratory rodents. The key target for PPs is the nuclear receptor peroxisome proliferator-activated receptor-alpha (PPARalpha) and these chemicals cause cancer by altering the expression of a subset of genes involved in cell growth regulation. The purpose of the present study was to utilize high-density gene expression arrays to examine the genes regulated by the potent PP Wy14,643 (50 microM, 6 h) in both rat (FaO) and human (HepG2) hepatoma cells. Treatment of FaO cells, but not HepG2, revealed the expected fatty acid catabolism genes. However, a larger than expected number of protein kinases, phosphatases, and signaling molecules were also affected exclusively in the FaO cells, including MAPK-phosphatase 1 (MKP-1), Janus-activated kinases 1 and 2 (JAK1 and 2), and glycogen synthetase kinase alpha and beta (GSKalpha and beta). The mRNA accumulation of these genes as well as the protein level for GSK3alpha, JAK1, and JAK2 and MKP-1 activity was corroborated. Due to the importance of MKP 1 in cell signaling, this induction was examined further and was found to be controlled, at least in part, at the level of the gene's promoter. Interestingly, overexpression of MKP-1 in turn affected the constitutive activity of PPARalpha. Taken together, the gene expression arrays revealed an important subset of PP regulated genes to be kinases and phosphatases. These enzymes not only would affect growth factor signaling and cell cycle control but also could represent feedback control mechanisms and modulate the activity of PPARalpha. PMID- 12729719 TI - Purification and characterization of a soluble form of rat liver NADPH-cytochrome P-450 reductase highly expressed in Escherichia coli. AB - A recombinant cDNA of rat liver NADPH-cytochrome P-450 reductase (CPR), which lacks the N-terminal hydrophobic region, was amplified by PCR and cloned. The N truncated cDNA named tCPR was ligated into a pBAce vector and expressed. The tCPR protein expressed in Escherichia coli was recovered into the soluble fraction of the cell lysate and purified to homogeneity by three sequential purification procedures; (I) anion-exchange chromatography on a DEAE-cellulose (DE-52) column, (II) affinity chromatography on 2('),5(')-ADP Sepharose 4B, and (III) chromatography on a hydroxyapatite column. The average yield was 47mg per liter of culture medium. The absorption spectrum of the purified tCPR protein was identical to that of a native full-length CPR purified from rat liver, indicating that tCPR also possesses one molecule each of FAD and FMN. The tCPR protein was able to reduce cytochrome c and was also able to assist heme degradation by a soluble form of rat heme oxygenase-1. However, it failed to support the O deethylation of 7-ethoxycoumarin by cytochrome P-450 1A1, indicating that the presence of the N-terminal hydrophobic domain is necessary for CPR to interact with cytochrome P-450. Previously, to prepare a soluble form of CPR, full-length CPR was treated with proteinases that selectively removed the N-terminal domain. With the expression system established in this study, however, the soluble and biologically active tCPR protein can be readily prepared in large amounts. This expression system will be useful for mechanistic as well as structural studies of CPR. PMID- 12729720 TI - Overexpression and reconstitution of a Rieske iron-sulfur protein from the higher plant. AB - The iron-sulfur protein subunit, known as the Rieske protein, is one of the central components of the cytochrome b(6)f complex residing in chloroplast and cyanobacterial thylakoid membranes. We have constructed plasmids for overexpression in Escherichia coli of full-length and truncated Rieske (PetC) proteins from the Spinacia oleracea fused to MalE. Overexpressed fusion proteins were predominantly found (from 55 to 70%) in cytoplasm in a soluble form. The single affinity chromatography step (amylose resine) was used to purify about 15mg of protein from 1 liter of E. coli culture. The isolated proteins were electrophoretically pure and could be used for further experiments. The NifS-like protein IscS from the cyanobacterium Synechocystis PCC 6803 mediates the incorporation of 2Fe-2S clusters into apoferredoxin and cyanobacterial Rieske apoprotein in vitro. Here, we used the recombinant IscS protein for the enzymatic reconstitution of the iron-sulfur cluster into full-length Rieske fusion and truncated Rieske fused proteins. Characterization by EPR spectroscopy of the reconstituted proteins demonstrated the presence of a 2Fe-2S cluster in both full length and truncated Rieske fusion proteins. PMID- 12729721 TI - Expression in E. coli and purification of Thermus thermophilus translation initiation factors IF1 and IF3. AB - The initiation of protein translation in bacteria requires in addition to mRNA, fMet-tRNA, and ribosomal subunits three protein factors, the initiation factor 1 (IF1), initiation factor 2 (IF2), and initiation factor 3 (IF3). The genes coding for IF1 and IF3 from Thermus thermophilus have been identified and cloned into pET expression vector and were expressed as soluble proteins in Escherichia coli. IF1 was purified by a DEAE-cellulose chromatography, followed by heat denaturation, chromatography on Hydroxylapatit, and gel permeation chromatography using Sephacryl 200HR. For the purification of IF3, a heat denaturation step is followed by anion-exchange chromatography on Q-Sepharose FF and gel permeation chromatography on Sephacryl 200HR. Using these procedures we obtained chromatographically pure and biologically active preparations of both T. thermophilus IF1 and IF3. PMID- 12729722 TI - Purification and characterization of recombinant protein kinase CK2 from Zea mays expressed in Escherichia coli. AB - Recombinant protein kinase subunits rmCK2alpha-1 and rmCK2beta-1 from Zea mays were expressed separately in Escherichia coli and assembled to a fully active tetrameric holoenzyme complex in vitro. The obtained maize holoenzyme was purified to homogeneity, biochemically characterized, and compared to CK2 from human. Kinetic measurements of the recombinant maize holoenzyme (rmCK2) revealed k(cat) values for ATP and GTP of 4 and 2s(-1), respectively; whereas the recombinant maize catalytic subunit showed almost equal values for ATP and GTP, i.e., ca. 0.8s(-1). A comparison of the k(cat)/K(m) ratio between the maize holoenzyme and the catalytic subunit from CK2 maize shows that the incorporation of the catalytic subunit into the holoenzyme leads to a 14-fold activation in the case of ATP and 8-fold activation in the case of GTP. The maize holoenzyme is about 10 times more sensitive towards CK2 inhibitor heparin, on the other hand, it is stimulated only 0% by polylysine as compared to the human counterpart. The maize holoenzyme activity is more sensitive towards NaCl concentrations higher than those of rhCK2 and treatment with urea showed that rmCK2 holoenzyme was denatured more readily than the human holoenzyme. PMID- 12729723 TI - An exotype alginate lyase in Sphingomonas sp. A1: overexpression in Escherichia coli, purification, and characterization of alginate lyase IV (A1-IV). AB - Sphingomonas sp. A1 (strain A1) cells contain three kinds of endotype alginate lyases [A1-I, A1-II, and A1-III], all of which are formed from a common precursor through posttranslational processing. In addition to these lyases, another type of lyase (A1-IV) that acts on oligoalginates exists in the bacterium. A1-IV was overexpressed in Escherichia coli cells through control of its gene under the T7 promoter. The expression level of the enzyme in E. coli cells was 8.6U/L-culture, which was about 270-fold higher than that in strain A1 cells. The enzyme was purified to homogeneity through three steps with an activity yield of 10.9%. The optimal pH and temperature, thermal stability, and mode of action of the purified enzyme were similar to those of the native enzyme from strain A1 cells. A1-IV exolytically degraded oligoalginates, which were produced from alginate through the reaction of A1-I, A1-II, or A1-III, into monosaccharides, indicating that the cooperative actions of these four enzymes cause the complete depolymerization of alginate in strain A1 cells. PMID- 12729724 TI - Expression and purification of recombinant Kaposi's sarcoma-associated herpesvirus DNA polymerase using a Baculovirus vector system. AB - The DNA polymerase (POL) of Kaposi's sarcoma-associated herpesvirus (KSHV) is essential for viral DNA replication and, thus, may be considered as a viable target for anti-KSHV therapeutics. To produce large quantities of homogeneous and pure POL required for high-throughput screening (HTS) for inhibitors, we generated a recombinant baculovirus vector encoding a hexahistidine (His6)-tagged POL and infected Spodoptera frugiperda Sf-9 insect cells. High expression of recombinant POL (rPOL) was achieved for up to 72h post-infection. The rPOL was solubilized in lysis buffer containing 0.3% Cymal-5 detergent, purified by metal chelating and dsDNA-cellulose affinity chromatography, and analyzed by anti-His antibody Western blot and mass spectrometry. The functionality of rPOL was confirmed by its DNA synthesis activity in vitro, which was effectively blocked by the anti-herpetic DNA polymerase inhibitors, foscarnet and cidofovir diphosphate, in a dose-dependent manner. The POL expressed and purified from the recombinant baculovirus-infected insect cells may be useful toward the development of HTS of large chemical libraries to identify novel KSHV DNA polymerase inhibitors. PMID- 12729725 TI - Cloning, expression, and characterization of diuretic hormone Manduca diuresin from Manduca sexta in Escherichia coli. AB - Manduca diuresin (MD), a 30 amino acid peptide isolated from the tobacco hornworm Manduca sexta, was found to play an important role in the regulation of water and salt balance in the insect. To facilitate studies relating to the function and structure of MD, a synthetic gene encoding MD was assembled and expressed in Escherichia coli. Using an excretion vector, expression of the MD gene in an induced transformant was detected at the transcriptional and translational levels by Northern-blot and ELISA analyses, respectively. With the use of glutathione-S transferase as the reporter protein, MD was confirmed to be expressed in E. coli. The recombinant product was resolved by reverse-phase HPLC into three peptide groups of different retention times, which were shown by mass spectrometry to be composed of MD deletants missing various lengths of the C-terminus. Despite the deletions and the absence of an amidated C-terminus, the deletants were shown to be biologically active, suggesting the importance of the N-terminus of MD for biological activity. PMID- 12729726 TI - Overexpression of hepatitis B virus surface antigens including the preS1 region in a serum-free Chinese hamster ovary cell line. AB - Current hepatitis B virus (HBV) vaccines consist of preparations of recombinant HBV major surface antigen (sAg) and are protective in about 90-95% of vaccinated subjects. In improved vaccines, the frequency of nonresponders to the classical vaccine could be reduced by including additional epitopes from the preS-domains of the middle and large surface antigens. In this report, the development and characterization of a CHO cell line for HBsAg, expressing major, middle, and large antigens are described. Despite the previously reported retention of secreted proteins by the preS1 domain, cell lines could be amplified that secreted large amounts of the complete set of antigens. A producer line was established that expressed 1mg HBsAg per 100ml suspension culture per week during exponential growth. The productivity per cell increased further by at least threefold when the culture reached the stationary phase at high cell densities. In the production cell line, several hundred copies of the HBV vector were integrated at two adjacent sites into chromosome 2. The cell line was adapted to growth in a defined protein-free medium minimizing the risk of adventitious agents introduced by animal derived supplements. The cell line stably produced antigen over several months. In the candidate vaccine, both preS2 and preS1 domains were present at ratios similar to HBsAg from human sera. In summary, a production cell line for an improved HBV vaccine is presented with properties such as high productivity, long term stability of expression, and growth in protein-free media. PMID- 12729727 TI - Chaperone-assisted expression, purification, and characterization of recombinant nitrile hydratase NI1 from Comamonas testosteroni. AB - Nitrile hydratases (NHases) are industrially important iron- and cobalt containing enzymes that are used in the large-scale synthesis of acrylamide. Heterologous expression of NHases has been complicated by the fact that other proteins (activators or metallochaperones) appear to be required to produce NHases in their catalytically active form. We report a novel heterologous system for the expression of catalytically active iron-containing NI1 NHase in Escherichia coli, involving coexpression with the E. coli GroES and GroEL chaperones. The purified recombinant enzyme was found to be highly similar to the enzyme purified from Comamonas testosteroni according to its spectroscopic features, catalytic properties with various substrates, and post-translational modifications. In addition, we report a rapid and convenient spectrophotometric method to monitor the activity of NI1 NHase during purification. PMID- 12729728 TI - A novel member of glycoside hydrolase family 88: overexpression, purification, and characterization of unsaturated beta-glucuronyl hydrolase of Bacillus sp. GL1. AB - Unsaturated beta-glucuronyl hydrolase of Bacillus sp. GL1 catalyzes the hydrolytic release of unsaturated glucuronic acids from oligosaccharides produced through the reactions of polysaccharide lyases such as gellan, xanthan, hyaluronate, and chondroitin lyases. An overexpression system for the enzyme was constructed in Escherichia coli cells involving regulation of the enzyme gene under the T7 promoter and terminator. The expression level of the enzyme in E. coli cells was 250-fold higher than that in Bacillus sp. GL1 cells. The enzyme expressed in E. coli cells was purified and characterized. The optimal pH and temperature, and substrate specificity of the purified enzyme were similar to those of the native enzyme from Bacillus sp. GL1 cells, although the enzyme expressed in E. coli cells underwent self-assembly into polymeric forms through the formation of intermolecular disulfide bonds. Circular dichroism analysis indicated that the secondary structure of the enzyme was rich in alpha-helices. Genes showing high identity (over 40% identity) with that of the enzyme were found in the genomes of some pathogenic bacteria, such as Streptococcus pyogenes and Streptococcus pneumoniae, which cause serious diseases (e.g., meningitis and pneumonia). Therefore, the enzyme of Bacillus sp. GL1 and the streptococcal proteins form a new glycoside hydrolase family, 88. PMID- 12729729 TI - Construction and characterization of a recombinant esterase with high activity and enantioselectivity to (S)-ketoprofen ethyl ester. AB - The ester-hydrolyzing enzyme families, including lipase and esterase, mediated a broad range of reactions and, thus, were able to act on a variety of ester compounds that are found naturally or exploited industrially. With the increasing demand for pharmacological use, attempts to produce an enantiomer (S)-ketoprofen from the corresponding ethyl ester have recently been proliferating, but information about the structure and function of related enzymes has not been reported to date in detail. Here, we reported the construction, expression, and one-step purification of a potential esterase in Escherichia coli with a hexahistidine tag at its N-terminus. The expression level of the enzyme was more than 20% of the total protein in E. coli, resulting in approximately 1.2mg of the purified proteins by an affinity resin, Ni-NTA, from a 0.2L of bacterial culture in a single step. As typical properties, its innate traits that revealed favorable reactions at alkaline pH and high activity to the triglycerides composed of short chain fatty acids (99% ee(p)). The small scale conversion using the recombinant enzyme strongly suggested the enzyme to be useful for enzyme-mediated chiral resolution of (S)-ketoprofen. PMID- 12729730 TI - Purified promyelocytic leukemia coiled-coil aggregates as a tetramer displaying low alpha-helical content. AB - The promyelocytic leukemia (PML) gene is involved in the 15/17 chromosomal translocation of acute promyelocytic leukemia (APL). It encodes a nuclear phosphoprotein containing an alpha-helical coiled-coil domain with four heptad repeats. The heptad repeats consist of four clusters of hydrophobic amino acids that mediate in vivo the complex formation between PML and other PML molecules or PML-RARalpha mutant protein. In this report, we show the production of PML coiled coil (fragment 223-360) as a fusion protein, its solubilization by the combined action of two different detergents, and its purification with affinity chromatography after column proteolytic cleavage. The FPLC chromatograms of the purified coiled-coils, carried out under non-denaturing conditions, show that the peptide elutes only in the presence of Sarkosyl detergent (conc. 0.1%) and, under these conditions, elutes as a tetrameric complex. This confirms the evidence from in vivo experiments that this region is responsible for protein complex formation. The HPLC analyses show the presence of a single peak eluting under highly hydrophobic conditions, indicating the high hydrophobicity of the peptide in accordance with the primary sequence analysis. Finally, the purified peptide was structurally characterized by means of circular dichroism (CD) measurements that were carried out with low Sarkosyl concentration (0.003%). The CD spectra indicate a low alpha-helical content (13.5%) with respect to predictions based on the primary sequence analysis (PSI-PRED, SS-PRO, and J-PRED), suggesting that the alpha-helix content could be modulated by coiled-coil surrounding domains and/or by other post-translational modifications, even if the effect of the Sarkosyl on the peptide secondary structure cannot be excluded. PMID- 12729731 TI - Expression in Pichia pastoris and purification of a membrane-acting immunotoxin based on a synthetic gene coding for the Bacillus thuringiensis Cyt2Aa1 toxin. AB - We explored the production in Pichia pastoris of a membrane-acting immunotoxin (IT) based on the Cyt2Aa1 toxin from the bacterium Bacillus thuringiensis subspecies kyushuensis. Initial attempts at the P. pastoris expression of Cyt2Aa1 were not successful due to the high A+T-content of the native bacterial gene, resulting in premature transcription termination. Accordingly, we designed and constructed a synthetic cyt2Aa1 gene (syncyt2Aa1)(2) that was optimised for expression in this eukaryotic host. This was achieved through a recursive PCR strategy where the overall G+C-content of the cyt2Aa1 DNA sequence was systematically increased to approximately 50% compared to approximately 30% in the native bacterial gene and only the P. pastoris preferred codons were used. A synthetic DNA sequence coding for a soluble and flexible serine/glycine linker was then used to genetically fuse syncyt2Aa1 with the human single-chain antibody fragment (scFv) C6.5 targeting p185(HER-2), a cell-surface glycoprotein overexpressed in 30% of human breast and ovarian cancers. Subsequent expression of the resulting IT construct [scFvC6.5-syncyt2Aa1(mychis(6))](2) led to high level accumulation of the recombinant protein in yeast membranes. Although the solubilisation of scFvC6.5-syncyt2Aa1(mychis(6)) from P. pastoris membranes necessitated the use of guanidine hydrochloride, the use of subsequent in vitro refolding and immobilised metal affinity chromatography (IMAC) steps allowed purification of the recombinant product at yields as high as approximately 10 mgl(-1) culture. Despite being core N-linked glycosylated and retaining part of the yeast secretion signal, the P. pastoris produced scFvC6.5 syncyt2Aa1(mychis(6)) exhibited significant specific activity for p185(HER-2) overexpressing SK-BR-3 cells but not p185(HER-2)-negative Swiss 3T3 cells or human erythrocytes. PMID- 12729732 TI - Characterization of factors favoring the expression of soluble protozoan tubulin proteins in Escherichia coli. AB - The alpha- and beta-tubulin genes of the parasitic protozoa Giardia duodenalis, Cryptosporidium parvum, and Encephalitozoon intestinalis have been overexpressed in soluble form using Escherichia coli-based expression systems. Several expression systems were compared in terms of the amount of soluble protein produced with different fusion partners, strains of E. coli BL21, and expression temperatures. The cleavability of the fusion partner was also assessed in terms of post-expression applications of the recombinant protein. The maltose-binding protein (MBP) and glutathione S-transferase (GST) fusion partners produced the highest expression levels for all six proteins without the formation of inclusion bodies. The expression system also provided a means of purifying the soluble protein using affinity and anion-exchange chromatography while minimizing protein losses. The yield and purity were therefore very high for both the MBP and GST systems. The tubulin monomers were demonstrated to be assembly-competent using a standard dimerization assay and also retained full antigenicity with monoclonal antibodies. This study presents several methods which are suitable for producing soluble tubulin monomers and, thus, circumventing the formation of inclusion bodies which necessitates re-folding of the tubulin. PMID- 12729734 TI - Low temperature and glucose enhanced T7 RNA polymerase-based plasmid stability for increasing expression of glucagon-like peptide-2 in Escherichia coli. AB - The high activity of T7 RNA polymerase has made the T7 RNA polymerase-based expression system very powerful for high-level expression of recombinant protein. However, the overactivity of T7 RNA polymerase would also bring about negative effects on plasmid stability and protein production, especially when expressing a toxic protein. If the latter role is dominant, it is necessary to adopt some measures to attenuate the activity or the amount of T7 RNA polymerase in the cells. Apart from the stringent regulation by inserting some genes reducing the amount or the activity of T7 RNA polymerase into plasmids, optimizing the culture conditions would be another way. In this work, we have studied the effects of various culture conditions on the plasmid stability and the target protein yield including selective pressure, culture temperature, toxicity of the target protein and the catabolite repression caused by glucose. The results have indicated that adding antibiotic after induction has little effect in increasing plasmid stability, but inducing expression at low temperature and adding glucose to the medium improved the plasmid stability and the protein yield to a large extent. PMID- 12729733 TI - Expression, purification, and initial characterization of a recombinant form of plant PEP-carboxylase kinase from CAM-induced Mesembryanthemum crystallinum with enhanced solubility in Escherichia coli. AB - Plant phosphoenolpyruvate-carboxylase kinase (PEPC-kinase [PpcK]) is the smallest Ser/Thr kinase identified to date, having a molecular mass of approximately 32,000. This novel, monomeric kinase is dedicated to the phosphorylation of plant PEPC, thereby regulating this target enzyme's activity and allosteric properties. Although several recombinant, non-fusion PpcK proteins have been produced recently in Escherichia coli, these are plagued by their high degree of insolubility. Here, we report the use of the native, E. coli NusA protein and a related E. coli expression vector (pET-43a(+) [Novagen]) for enhancing the solubility of this recalcitrant Ser/Thr kinase at least 10-fold by its production as a dual 6xHis-tagged NusA/McPpcK1 fusion protein, which accounts for approximately 10% of the soluble protein fraction from induced cells. Capture of this fusion protein from the centrifuged cell extract by immobilized metal (Ni(2+)) affinity-chromatography, its "on-bead" cleavage by thrombin, and subsequent elution yielded milligram quantities of a "free," approximately 36-kDa form of PpcK for further purification by fast-protein liquid chromatography on blue dextran-agarose or preparative SDS-PAGE. Steady-state kinetic analysis of the former, active preparation revealed that this dedicated kinase discriminates against neither various isoforms of plant PEPC nor certain mutant forms of recombinant C(4) PEPC. Alternatively, the latter, electrophoretically homogeneous sample of the approximately 36-kDa polypeptide was used as antigen for polyclonal antibody production in rabbits. The antibodies against the recombinant McPpcK1 from Mesembryanthemum crystallinum cross-reacted on Western blots with an enriched preparation of the maize-leaf kinase, but not with the parent crude extract, thus directly documenting this protein's extremely low abundance in vivo. However, these antibodies were effective in immunoprecipitating 32P-based PpcK activity from crude, desalted extracts of maize leaves and soybean root nodules. PMID- 12729735 TI - Repression of the TMEFF2 promoter by c-Myc. AB - TMEFF2 is a novel transmembrane protein, containing two follistatin domains and an epidermal growth factor-like motif that is mainly expressed in the prostate and brain. Recently, we showed that expression of TMEFF2 could inhibit prostate cancer cell growth. In addition, the TMEFF2 gene is frequently hypermethylated in human tumor cells, suggesting that it might be a tumor suppressor gene. We cloned the 5'-flanking region of the human TMEFF2 gene and using a luciferase reporter assay showed that it contains a functional promoter. The 0.7 kb region upstream to the TMEFF2 transcription start site encompasses the minimal promoter required for TMEFF2 expression. Sequence analysis of the TMEFF2 promoter revealed potential binding sites for several transcription factors including Sp1 and an E box that could be recognized by c-Myc. An inverse correlation between TMEFF2 and c-Myc expression was found in CWR22 prostate xenografts. Reporter gene and mobility shift assays demonstrated that c-Myc could repress TMEFF2 gene expression through its cognate site. In light of the probable role of TMEFF2 in inhibiting cell growth, its suppression may contribute to the oncogenic properties of c-Myc. PMID- 12729736 TI - Crystal structure of the human CCA-adding enzyme: insights into template independent polymerization. AB - All tRNA molecules carry the invariant sequence CCA at their 3'-terminus for amino acid attachment. The post-transcriptional addition of CCA is carried out by ATP(CTP):tRNA nucleotidyltransferase, also called CCase. This enzyme catalyses a unique template-independent but sequence-specific nucleotide polymerization reaction. In order to reveal the molecular mechanism of this activity, we solved the crystal structure of human CCase by single isomorphous replacement. The structure reveals a four domain architecture with a cluster of conserved residues forming a positively charged cleft between the first two domains. Structural homology of the N-terminal CCase domain to other nucleotidyltransferases could be exploited for modeling a tRNA-substrate complex. The model places the tRNA 3'-end into the N-terminal nucleotidyltransferase site, close to a patch of conserved residues that provide the binding sites for CTP and ATP. Based on our results, we introduce a corkscrew model for CCA addition that includes a fixed active site and a traveling tRNA-binding region formed by flexible parts of the protein. PMID- 12729737 TI - Towards understanding human mitochondrial leucine aminoacylation identity. AB - Specific recognition of tRNAs by aminoacyl-tRNA synthetases is governed by sets of aminoacylation identity elements, well defined for numerous prokaryotic systems and eukaryotic cytosolic systems. Only restricted information is available for aminoacylation of human mitochondrial tRNAs, despite their particularities linked to the non-classical structures of the tRNAs and their involvement in a growing number of human neurodegenerative disorders linked to mutations in the corresponding tRNA genes. A major difficulty to be overcome is the preparation of active in vitro transcripts enabling a rational mutagenic analysis, as is currently performed for classical tRNAs. Here, structural and aminoacylation properties of in vitro transcribed tRNA(Leu(UUR)) are presented. Solution probing using a combination of enzymatic and chemical tools revealed only partial folding into an L-shaped structure, with an acceptor branch but with a floppy anticodon branch. Optimization of aminoacylation conditions allowed charging of up to 75% of molecules, showing that, despite its partially relaxed structure, in vitro transcribed tRNA(Leu(UUR)) is able to adapt to the synthetase. In addition, mutational analysis demonstrates that the discriminator base as well as residue A14 are important leucine identity elements. Thus, human mitochondrial leucylation is dependent on rules similar to those that apply in Escherichia coli. The impact of a subset of pathology-related mutations on aminoacylation and on tRNA structure, has been explored. These variants do not show significant structural rearrangements and either do not affect aminoacylation (mutations T3250C, T3271C, C3303T) or lead to marked effects. Interestingly, two variants with a mutation at the same position (A3243G and A3243T) lead to markedly different losses in aminoacylation efficiencies (tenfold and 300-fold, respectively). PMID- 12729738 TI - Exploration of the transition state for tertiary structure formation between an RNA helix and a large structured RNA. AB - Docking of the P1 duplex into the pre-folded core of the Tetrahymena group I ribozyme exemplifies the formation of tertiary interactions in the context of a complex, structured RNA. We have applied Phi-analysis to P1 docking, which compares the effects of modifications on the rate constant for docking (k(dock)) with the effects on the docking equilibrium (K(dock)). To accomplish this we used a single molecule fluorescence resonance energy transfer assay that allows direct determination of the rate constants for formation of thermodynamically favorable, as well as unfavorable, states. Modification of the eight groups of the P1 duplex that make tertiary interactions with the core and changes in solution conditions decrease K(dock) up to 500-fold, whereas k(dock) changes by NADP(+)+2 GSH) has become an attractive drug target for antimalarial drug development. Here, we report the 2.6A resolution crystal structure of P.falciparum GR. The homodimeric flavoenzyme is compared to the related human GR with focus on structural aspects relevant for drug design. The most pronounced differences between the two enzymes concern the shape and electrostatics of a large (450A(3)) cavity at the dimer interface. This cavity binds numerous non-competitive inhibitors and is a target for selective drug design. A 34-residue insertion specific for the GRs of malarial parasites shows no density, implying that it is disordered. The precise location of this insertion along the sequence allows us to explain the deleterious effects of a mutant in this region and suggests new functional studies. To complement the structural comparisons, we report the relative susceptibility of human and plasmodial GRs to a series of tricyclic inhibitors as well as to peptides designed to interfere with protein folding and dimerization. Enzyme-kinetic studies on GRs from chloroquine-resistant and chloroquine-sensitive parasite strains were performed and indicate that the structure reported here represents GR of P.falciparum strains in general and thus is a highly relevant target for drug development. PMID- 12729761 TI - The structure of holo and metal-deficient wild-type human Cu, Zn superoxide dismutase and its relevance to familial amyotrophic lateral sclerosis. AB - Cu, Zn superoxide dismutase (SOD1) forms a crucial component of the cellular defence against oxidative stress. Zn-deficient wild-type and mutant human SOD1 have been implicated in the disease familial amyotrophic lateral sclerosis (FALS). We present here the crystal structures of holo and metal-deficient (apo) wild-type protein at 1.8A resolution. The P21 wild-type holo enzyme structure has nine independently refined dimers and these combine to form a "trimer of dimers" packing motif in each asymmetric unit. There is no significant asymmetry between the monomers in these dimers, in contrast to the subunit structures of the FALS G37R mutant of human SOD1 and in bovine Cu,Zn SOD. Metal-deficient apo SOD1 crystallizes with two dimers in the asymmetric unit and shows changes in the metal-binding sites and disorder in the Zn binding and electrostatic loops of one dimer, which is devoid of metals. The second dimer lacks Cu but has approximately 20% occupancy of the Zn site and remains structurally similar to wild-type SOD1. The apo protein forms a continuous, extended arrangement of beta-barrels stacked up along the short crystallographic b-axis, while perpendicular to this axis, the constituent beta-strands form a zig-zag array of filaments, the overall arrangement of which has a similarity to the common structure associated with amyloid-like fibrils. PMID- 12729763 TI - Insights into the catalytic mechanism of cofactor-independent phosphoglycerate mutase from X-ray crystallography, simulated dynamics and molecular modeling. AB - Phosphoglycerate mutases catalyze the isomerization of 2 and 3-phosphoglycerates, and are essential for glucose metabolism in most organisms. Here, we further characterize the 2,3-bisphosphoglycerate-independent phosphoglycerate mutase (iPGM) from Bacillus stearothermophilus by determination of a high-resolution (1.4A) crystal structure of the wild-type enzyme and the crystal structure of its S62A mutant. The mutant structure surprisingly showed the replacement of one of the two catalytically essential manganese ions with a water molecule, offering an additional possible explanation for its lack of catalytic activity. Crystal structures invariably show substrate phosphoglycerate to be entirely buried in a deep cleft between the two iPGM domains. Flexibility analyses were therefore employed to reveal the likely route of substrate access to the catalytic site through an aperture created in the enzyme's surface during certain stages of the catalytic process. Several conserved residues lining this aperture may contribute to orientation of the substrate as it enters. Factors responsible for the retention of glycerate within the phosphoenzyme structure in the proposed mechanism are identified by molecular modeling of the glycerate complex of the phosphoenzyme. Taken together, these results allow for a better understanding of the mechanism of action of iPGMs. Many of the results are relevant to a series of evolutionarily related enzymes. These studies will facilitate the development of iPGM inhibitors which, due to the demonstrated importance of this enzyme in many bacteria, would be of great potential clinical significance. PMID- 12729764 TI - What is the minimum number of letters required to fold a protein? AB - Experimental studies have shown that the full sequence complexity of naturally occurring proteins is not required to generate rapidly folding and functional proteins, i.e. proteins can be designed with fewer than 20 letters. This raises the question of what is the minimum number of amino acid types required to encode complex protein folds? Here, we investigate this issue from three aspects. First, we study the minimum sequence complexity that can reserve the necessary structural information for detection of distantly related homologues. Second, we compare the ability of designing foldable model sequences over a wide range of reduced amino acid alphabets, which find the minimum number of letters that have the similar design ability as 20. Finally, we survey the lower bound of alphabet size of globular proteins in a non-redundant protein database. These different approaches give a remarkably consistent view, that the minimum number of letters required to fold a protein is around ten. PMID- 12729766 TI - A kinetic mechanism for the fast movement of Chara myosin. AB - Endoplasmic streaming of characean cells of Nitella or Chara is known to be in the range 30-100 microm/second. The Chara myosin extracted from the cells and fixed onto a glass surface was found to move muscle actin filaments at a velocity of 60 microm/second. This is ten times faster than that of skeletal muscle myosin (myosin II). In this study, the displacement caused by single Chara myosin molecules was measured using optical trapping nanometry. The step size of Chara myosin was approximately 19nm. This step size is longer than that of skeletal muscle myosin but shorter than that of myosin V. The dwell time of the steps was relatively long, and this most likely resulted from two rate-limiting steps, the dissociation of ADP and the binding of ATP. The rate of ADP release from Chara myosin after the completion of the force-generation step was similar to that of myosin V, but was considerably slower than that of skeletal muscle myosin. The 19nm step size and the dwell time obtained could not explain the fast movement. The fast movement could be explained by the load-dependent release of ADP. As the load imposed on the myosin decreased, the rate of ADP release increased. We propose that the interaction of Chara myosin with an actin filament resulted in a negative load being imposed on other myosin molecules interacting with the same actin filament. This resulted in an accelerated release of ADP and the fast sliding movement. PMID- 12729765 TI - Structural and functional defects caused by point mutations in the alpha crystallin domain of a bacterial alpha-heat shock protein. AB - The diverse family of alpha-crystallin-type small heat shock proteins (alpha-Hsps or sHsps) is characterised by a central, moderately conserved alpha-crystallin domain. Oligomerisation followed by dissociation of subparticles is thought to be a prerequisite for chaperone function. We demonstrate that HspH, a bacterial alpha-Hsp from the soybean-symbiont Bradyrhizobium japonicum, assembles into dynamic complexes freely exchanging subunits with homologous and heterologous complexes. The importance of the alpha-crystallin domain for oligomerisation and chaperone activity was tested by site-directed mutagenesis of 12 different residues. In contrast to mammalian alpha-Hsps, the majority of these mutations elicited severe structural and functional defects in HspH. The individual exchange of five amino acid residues throughout the alpha-crystallin domain was found to compromise oligomerisation to various degrees. Assembly defects resulting in complexes of reduced size correlated with greatly decreased or abolished chaperone activity, reinforcing that complete oligomerisation is required for functionality. Mutation of a highly conserved glycine (G114) at the C-terminal end of the alpha-crystallin domain specifically impaired chaperone activity without interfering with oligomerisation properties, indicating that this residue is critical for substrate interaction. The structural and functional importance of this and other residues is discussed in the context of a modeled three-dimensional structure of HspH. PMID- 12729767 TI - Identification of a key structural element for protein folding within beta hairpin turns. AB - Specific residues in a polypeptide may be key contributors to the stability and foldability of the unique native structure. Identification and prediction of such residues is, therefore, an important area of investigation in solving the protein folding problem. Atypical main-chain conformations can help identify strains within a folded protein, and by inference, positions where unique amino acids may have a naturally high frequency of occurrence due to favorable contributions to stability and folding. Non-Gly residues located near the left-handed alpha helical region (L-alpha) of the Ramachandran plot are a potential indicator of structural strain. Although many investigators have studied mutations at such positions, no consistent energetic or kinetic contributions to stability or folding have been elucidated. Here we report a study of the effects of Gly, Ala and Asn substitutions found within the L-alpha region at a characteristic position in defined beta-hairpin turns within human acidic fibroblast growth factor, and demonstrate consistent effects upon stability and folding kinetics. The thermodynamic and kinetic data are compared to available data for similar mutations in other proteins, with excellent agreement. The results have identified that Gly at the i+3 position within a subset of beta-hairpin turns is a key contributor towards increasing the rate of folding to the native state of the polypeptide while leaving the rate of unfolding largely unchanged. PMID- 12729769 TI - Advanced technology and the future of facial prosthetics in head and neck reconstruction. PMID- 12729768 TI - Hydration and packing are crucial to amyloidogenesis as revealed by pressure studies on transthyretin variants that either protect or worsen amyloid disease. AB - The formation of amyloid aggregates is the hallmark of the amyloidogenic diseases. Transthyretin (TTR) is involved in senile systemic amyloidosis (wild type protein) and familial amyloidotic polyneuropathy (point mutants). Through the use of high hydrostatic pressure (HHP), we compare the stability among wild type (wt) TTR, two disease-associated mutations (V30M and L55P) and a trans suppressor mutation (T119M). Our data show that the amyloidogenic conformation, easily populated in the disease-associated mutant L55P, can be induced by a cycle of compression-decompression with the wt protein rendering the latter highly amyloidogenic. After decompression, the recovered wt structure has weaker subunit interactions (loosened tetramer, T(4)(*)) and presents a stability similar to L55P, suggesting that HHP induces a defective fold in the wt protein, converting it to an altered conformation already present in the aggressive mutant, L55P. On the other hand, glucose, a chemical chaperone, can mimic the trans-suppression mutation by stabilizing the native state and by decreasing the amyloidogenic potential of the wt TTR at pH 5.0. The sequence of pressure stability observed was: L55P 0.05), the used technique of "RIF LF-I impaction and RIF BSSO advancement" tended to render excellent postsurgical stability in the horizontal (0.1+/-0.8mm mean posterior movement) and vertical (0.1+/-0.5mm mean inferior movement) direction. There was no instance of maxillary relapse of >2mm. Regarding mandibular BSSO advancement, the point B showed a significant vertical upward movement (1.6+/ 1.2mm) (P< 0.001) and a slight horizontal forward movement (0.3+/-2.0mm) (P> 0.05) at 1-year follow-up. The incidence of posterior relapse of >2mm accounted for 11.5%. The data confirm the concept that the bimaxillary approach of "LF-I impaction and BSSO advancement" using the described technique of RIF is a stable procedure in the treatment of open bite patients classified as vertical maxillary excess in combination with mandibular deficiency. PMID- 12729773 TI - Synovial chondromatosis of the temporomandibular joint: clinical, surgical and histological aspects. AB - Nine patients with histologically confirmed unilateral synovial chondromatosis of the temporomandibular joint were treated surgically with extirpation of loose bodies and partial synovectomy. In six of them the histological material was available for a systematic examination. The results of treatment were evaluated clinically and with MRI after a follow-up ranging between 1 and 17 years. Our findings suggest that synovial chondromatosis of the temporomandibular affects only the synovial lining of the upper compartment. The histological appearance is that of a benign chronic inflammation varying in severity and with metaplastic activity. The most specific clinical sign of synovial chondromatosis is swelling over the joint. Distension of the lateral capsule and fluid in the joint on the MRI are very suggestive of this diagnosis. Loose bodies also indicate synovial chondromatosis, but they are not always detected on the preoperative MRI. The surgical treatment should be conservative and include thorough removal of the loose bodies and partial synovectomy in areas with marked inflammation. PMID- 12729774 TI - Miniplate removal in trauma and orthognathic surgery--a retrospective study. AB - Records of patients undergoing the surgical removal of miniplates placed during the management of maxillofacial trauma (n=49) and orthognathic surgery (n=16) in a single unit, over a 2-year period have been analysed. Data concerning indications for plating, age and sex distribution, site of plating, time between insertion and removal, antibiotic prophylaxis, general medical factors and clinical indications for plate removal were evaluated for all patients. In addition, data for trauma patients included the site of fracture, and time delay between injury and plate insertion. Infection and/or wound dehiscence were the predominant causes for plate removal in both groups, in spite of the routine use of prophylactic antibiotics in a predominantly young, healthy cohort of patients.A removal rate of approximately 10% was found in the two groups of patients. This low rate would seem to imply that the routine removal of miniplates is not clinically indicated. PMID- 12729775 TI - Do frozen sections help achieve adequate surgical margins in the resection of oral carcinoma? AB - This retrospective study examined the notes of 82 patients who underwent an attempted curative resection of oro-pharyngeal carcinoma, and who had frozen sections taken, over a four year period. Three hundred and fifty mucosal, 179 deep tissue, and 22 nerve frozen sections had been taken. Concordance between cryostat and paraffin sections was 99.5% although no false positives or negatives for invasive tumour were found. Nine (11%) patients who had frozen sections which were reported as dysplastic or positive for invasive tumour underwent further local resection; excision was then found to be complete in 8 (10%) of these patients with further frozen sections. In 15 patients the margins of the main resection specimen were judged to be close to the tumour (<5mm), 5 patients had squamous epithelial dysplasia and 12 patients had invasive tumour at a resection margin. Ten of the 12 patients with margins containing invasive tumour had negative cryostat sections intraoperatively, which demonstrated problems with sampling which is the major drawback. During follow up, 14 patients developed local recurrence. Only one of the patients who had frozen section guided further local resection, developed local recurrence. Frozen sections help achieve tumour clearance. PMID- 12729776 TI - The radial free forearm flap--prelaminated versus non-prelaminated: a comparison of two methods. AB - In the present work two methods of harvesting the RFFF (radial free forearm flap) are compared: the prelaminated fasciomucosal and the non-prelaminated fasciocutaneous version. The flaps were employed for intraoral reconstruction after radical oncological surgery of the oral cavity. In most cases a squamous cell carcinoma was the present malignant tumour. A total of 32 patients were included in the study, 16 in each group. The design was retrospective. Data were obtained by clinical follow up, chart review and compiled database. Analysis included the amount of shrinkage of the flaps during the follow-up period, early postoperative complications, following function improving operations and early and late donor site difficulties. Prelaminated fasciomucosal flaps showed a higher shrinkage rate than fasciocutaneous non-prelaminated flaps. Due to this circumstance the number of following function improving operations was higher in the prelaminated flap group. Early wound healing difficulties were also seen more often in the prelaminated flap group, whereas donor site problems occurred more frequently in the non-prelaminated group. Mucosal prelamination of the RFFF is a promising method for a most physiological reconstruction of intraoral defects resulting from tumour surgery. Postoperative shrinkage is a problem in prelaminated RFFF. Our aim is to improve the prelamination technique in order to prevent shrinkage. The almost complete absence of donor site difficulties in prelaminated RFFF may represent a solution to this common and yet unsolved problem. PMID- 12729777 TI - Survival in patients under 45 years with squamous cell carcinoma of the oral tongue. AB - Squamous cell cancer among young adults is rare and thought to have aggressive biological behaviour and poor prognosis. Clinical case records of 76 patients under the age of 45 years treated for squamous cell carcinoma (SCC) of the oral tongue were retrospectively analysed to calculate the survival rates and the predictors of survival. The overall survival (OS) and disease free survival (DFS) at 5 year were 78.0% and 57.4% respectively. Tumour status, pathological node status, surgical clearance, the selection of appropriate treatment, type of primary and neck surgery, were found to significantly influence the survival rates. Young patients with SCC of the oral tongue were seen to have a comparable outcome and prognosis compared to that in older patients. PMID- 12729778 TI - Hypercalcemic complication in patients with oral squamous cell carcinoma. AB - Hypercalcemia is one of the metabolic complications associated with cancer. To assess the frequency of hypercalcemia in patients with squamous cell carcinoma (SCC), 242 patients who were evaluated as having SCC in the oral cavity between July 1995 and June 2001 were investigated. All patients were periodically monitored for their serum level of calcium (Ca). Hypercalcemia was defined as a serum Ca concentration higher than 11 mg/dl. By this definition, hypercalcemia was detected in 12 of the 242 patients (5.0%). All 12 patients were at an advanced stage of oral SCC. In these 12 patients, the serum level of parathyroid hormone-related protein (PTH-rP) was also significantly elevated. Therefore, we diagnosed these diseases as humoral hypercalcemia of malignancy (HHM). Moreover, we studied the efficacy of anti-hypercalcemic therapy on the quality of life (QOL). The European Organization for Research and Treatment of Cancer (EORTC) QLQ C30 was used for estimation of QOL. The patients with HHM who were administrated drugs such as bisphosphonate and calcitonin showed a reduction in their Ca and PTH-rP levels, and the six of ten EORTC QLQ-C30 subscales (emotional functioning, cognitive functioning, fatigue, dyspnoea, nausea/vomiting and appetite loss) were also improved after the anti-hypercalcemic therapy. However, these suppressive effects were temporary. The median survival time after the diagnosis of HHM was only 54.9+/-18.3 days (range 27-86 days). Therefore, HHM in SCC appears to be an ominous prognostic sign. Although anti-hypercalcemic therapy has a palliative role, the patients may be in less discomfort during the terminal stage of their illness. PMID- 12729780 TI - Transfusion transmitted virus (TTV) in dental patients. AB - Transfusion transmitted virus (TTV) is a new DNA virus found in patients with post-transfusion hepatitis. The prevalence of this virus among dental patients has not been reported, therefore, the prevalence of TTV infection in consecutive dental inpatients was evaluated. TTV DNA was assayed by the polymerase chain reaction (PCR) in 441 dental inpatients with oral cancer (n=192) or oral cysts (n=249). The serum HBs antigen and HCV antibody as well as aspartate transaminase (AST), alanine transaminase (ALT), and gamma glutamyl transpeptidase (gamma-GTP) concentrations were also measured. Of 441 subjects, 137 were infected with TTV (31.1%). This prevalence of TTV was much higher than that of HBV or HCV (HBV 1.2%; HCV 6.0%) in these dental patients. There was no gender or age difference in the prevalence of TTV infection. Of the 192 patients with oral cancer, 57 subjects had TTV in their sera, while 80 of 249 with oral cystic disease had TTV. The prevalence of TTV was similar between the two different disease groups. Neither the serum ALT nor serum AST concentrations were different between the subjects positive and negative for TTV DNA. In hospitalized dental patients, 31.1% were infected with TTV. The prevalence of TTV was much higher than that of HBV or HCV. There was no difference in the prevalence of TTV between subjects with cancer and cysts. Dentists should maintain high standards of infection control when treating any dental patient. PMID- 12729779 TI - Lack of association between hepatitis C virus and oral epithelial dysplasia in British patients. AB - Oral lichen planus may be a premalignant condition. An association between hepatitis C virus (HCV) infection and oral lichen planus has been described in Southern European and Japanese patients, and recently an association between HCV and oral squamous cell carcinoma was suggested from a study of Japanese patients. The present study investigated the frequency of chronic liver disease and HCV infection in UK patients with oral epithelial dysplasia (OED), a known premalignant disorder. Subjects included 75 patients with histologically proven OED and 110 healthy controls. Liver function and IgG antibodies to HCV were examined serologically. No patient with OED or control subject had serological evidence of hepatic disease, and anti-HCV antibodies were detected in only two (2.6%) of the 75 patients with OED, none of the controls being HCV seropositive. It is concluded that in the UK there is no association between HCV infection and OED. PMID- 12729781 TI - Intraoral grafting of an ex vivo produced oral mucosa equivalent: a preliminary report. AB - The objective of this study was to assess the efficacy of the use of an ex vivo produced oral mucosa equivalent (EVPOME) for intraoral grafting procedures. Autogenous keratinocytes were harvested from a punch biopsy 4 weeks prior to surgery, placed in a serum-free culture system and seeded onto a human cadaveric dermal equivalent, AlloDerm. Thirty patients with either a premalignant or cancerous lesion were triaged into two groups, depending on the stage of disease: Group 1: EVPOME or Group 2: AlloDerm, control without an epithelial layer. Clinically, EVPOME grafts were easy to handle and showed excellent compliance on grafting. Both, EVPOME and AlloDerm grafts, showed a 100% take rate. At 6 days post-grafting, the EVPOME clinically showed changes indicating vascular ingrowth and had cytologic evidence of the persistence of grafted cultured keratinocytes on the surface. The EVPOME grafts had enhanced maturation of the underlying submucosal layer associated with rapid epithelial coverage when compared to the AlloDerm grafts at biopsies taken at 28 days post-grafting. In summary, EVPOME appears to be an acceptable oral mucosal substitute for human intraoral grafting procedures and results in a more favorable wound healing response than AlloDerm alone. PMID- 12729782 TI - Search for deletion 22q11.2 in interphase nuclei of buccal mucosa of patients ascertained by isolated cleft palate: a new diagnostic approach. AB - A new approach for the detection of chromosome deletion 22q11.2 in interphase nuclei from buccal mucosa cells obtained by a non-invasive procedure is described. FISH analysis has been performed on samples from a group of 101 patients that presented consecutively for speech therapy and/or surgical correction of cleft palate. A normal result has been obtained in 98 patients; a deletion 22q11.2 was present in three patients (2.8%) with cleft palate. PMID- 12729783 TI - Dental and maxillofacial injuries among older New Zealanders during the 1990s. AB - Not only is the number of older people increasing in New Zealand, but a greater number of them are keeping their own teeth, with predictable consequences for the costs of treating (and rehabilitating) dental and maxillofacial trauma in that group. The aim of the study was to describe the occurrence of dental and maxillofacial trauma involving New Zealanders aged 65 and over during the 1990s. The investigation was a secondary analysis of routinely collected, national-level compensation and hospital treatment data. The analysis showed that: (1) there was an increase in the rate and absolute number of injuries among older people; (2) the dental trauma rate was highest among males in the youngest age group, while the facial fracture rate was highest among older females; and (3) there was a general increase in the contribution of falls to the occurrence of trauma. Measures which aim to reduce the occurrence of falls among older people will also reduce the oral and maxillofacial trauma rate in that age group. PMID- 12729784 TI - Incidence and pattern of maxillofacial fractures in the elderly. AB - The results of an audit analysing patients above 60 years and above treated for maxillofacial fractures over a 15-year period at the University College Hospital, lbadan are presented. Of the total number of 1689 patients of all ages treated for facial fractures, 53 (3.1%) were within the age range of the study. The male to female ratio was 1.1:1 and 41 (77.4%) patients were younger than 70 years. Road traffic accidents were the most common cause of facial fractures (58.5%) and the mandible was involved in 91% of cases. Complications were noted in 2 (3.8%) patients. Attention should be paid to update driving courses and home safety instructions in order to reduce the incidence of maxillofacial fractures in the elderly. PMID- 12729785 TI - Maxillofacial war injuries during the Iraq-Iran War: an analysis of 300 cases. AB - The following study focuses on three hundred maxillofacial war casualties that were admitted to the Basra Republic Hospital during the Iraq-Iran War. These three hundred cases were chosen on the basis of them being only oral and maxillofacial injuries. Of these cases, there was no mortality recorded. This was in part due to the rapid evacuation, immediate resuscitation and proper management of the casualties. An appropriately staffed hospital, efficient surgical techniques and the presence of highly skilled specialists working as a team also made this result possible. This analysis categorizes, evaluates and discusses the treatment of the casualties based on the severity of the injury and outlines several techniques used to treat such patients. PMID- 12729787 TI - Ectopic lingual thyroid: a case report. AB - A rare case of lingual thyroid of massive size is being reported in a 28-year-old female patient who presented with dysphonia, fullness in the throat, dysphagia and breathing difficulties. Fibre-optic nasopharyngoscopy revealed a sessile mass occupying the whole of the oro and hypopharynx. The mass was excised and histopathological examination was reported as thyroid tissue. Postoperatively the patient remained euthyroid and did not require thyroxine supplements since the hypoplastic thyroid gland in the prelaryngeal region remained functional. PMID- 12729786 TI - Dislocation of the mandibular condyle into the middle cranial fossa: report of a case with 5 year CT follow-up. AB - A 17-year-old patient suffered a dislocation of the right mandibular condyle into the middle cranial fossa following a motorcycle accident. Reduction was delayed because of serious orthopaedic injuries. The condyle was relocated into the glenoid fossa through a combined neurosurgical and preauricular approach. Five years later the patient presented for removal of wisdom teeth and a CT-scan was done to determine the effects of the trauma and management to the condyle. A fibro-osseous ankylosis had occurred in the right temporomandibular joint. The radiological changes confirm that early management appears to improve the prognosis for these injuries. PMID- 12729788 TI - Vanishing bone disease in a five year old: report of a case and review of the literature. AB - Vanishing bone disease is a rare condition of unknown aetiology. It can affect almost any bone, including those of the maxillofacial region. It is most commonly seen in the second and third decades of life. To the author's knowledge, this is the second case reported in the maxillofacial region of a child within the first decade of life, and the first who survived. PMID- 12729790 TI - Profilin I colocalizes with speckles and Cajal bodies: a possible role in pre mRNA splicing. AB - Profilin is one of the major components controlling actin polymerization. Here, profilin I was located in fibroblasts and HeLa cells by the use of two different sets of affinity-purified antibodies. Both antibody preparations labeled nuclei in a speckle-like pattern and displayed extensive colocalization with small nuclear ribonucleoprotein particle (snRNP)-core proteins and p80 coilin containing Cajal bodies. Treatment with actinomycin D led to largely similar reorganizations of snRNPs and profilin, while profilin and Cajal bodies separated under these conditions. One of the profilin antibodies interfered with pre-mRNA splicing in vitro, further indicating a role for profilin during pre-mRNA processing. PMID- 12729789 TI - CD98-dependent homotypic aggregation is associated with translocation of protein kinase Cdelta and activation of mitogen-activated protein kinases. AB - CD98 is a protein found on the surface of many activated cell types, and is implicated in the regulation of cellular differentiation, adhesion, growth, and apoptosis. Despite many studies addressing CD98 function, there is little information on the intracellular signalling pathways that mediate its activity. In this study, we examine protein kinase pathways that are activated following ligation by the CD98 antibody AHN-18, an antibody that induces U937 homotypic aggregation and inhibits antigen presenting activity and T-cell activation. Ligation by CD98 antibody AHN-18 induces tyrosine kinase activity, but inhibition of this activity does not affect U937 aggregation. Ligation also induces membrane translocation of the serine/threonine kinase novel PKCdelta, but not other members of the PKC family. Translocation is blocked by rottlerin, and this inhibitor also blocks aggregation. PKCdelta activation in turn mediates activation of ERK1/2 and p38, as well as tyrosine phosphorylation of multiple proteins, and MAPK activation is essential for cellular aggregation. One of the targets of CD98-induced tyrosine phosphorylation is itself PKCdelta, suggesting that this phosphorylation may act as a negative feedback to limit the overall activation of the CD98 pathway. PMID- 12729791 TI - PKCeta associates with cyclin E/Cdk2 complex in serum-starved MCF-7 and NIH-3T3 cells. AB - Protein kinase C (PKC) encodes a family of enzymes implicated in cellular differentiation, growth control, and tumor promotion. However, very little is known with respect to the molecular mechanisms that link protein kinase C to cell cycle control. Here we report that PKCeta associates with the cyclin E/Cdk2 complex. This is shown for the ectopically overexpressed PKCeta in NIH-3T3 cells, the inducibly expressed PKCeta in MCF-7 cells (under control of the tetracycline responsive promoter), and the endogenously expressed PKCeta in mouse mammary epithelial HC11 cells. Subcellular cell fractionation experiments revealed that the complex with cyclin E is formed mostly in the nuclear fractions, although in these cells PKCeta is predominantly expressed in the cytosolic fractions. The complex of PKCeta and cyclin E was studied at various phases of the cell cycle, in serum-starved quiescent cells and in cells stimulated with serum to reenter the cell cycle. Interestingly, the interaction between PKCeta and cyclin E was most prominent in serum-starved cells and was disintegrated when cells entered the cells cycle. Immunofluorescence staining demonstrated that in serum-starved cells PKCeta is concentrated at the perinuclear zone, which is also the site of its colocalization with cyclin E. Colocalization of PKCeta and cyclin E in the perinuclear region was observed in serum-starved cells, and less in proliferating cells. These experiments suggest that the interaction between PKCeta and cyclin E is carefully regulated, and is correlated with the inactivated form of the cyclin E/Cdk2 complex. Thus, our studies support an important link between PKC and cell cycle control. PMID- 12729792 TI - In vitro aging of rat lung cells. Downregulation of telomerase activity and continuous decrease of telomere length are not incompatible with malignant transformation. AB - Most normal mammalian somatic cells cultivated in vitro enter replicative senescence after a finite number of divisions, as a consequence of the progressive shortening of telomeres during proliferation that reflects one aspect of organism/cellular aging. The situation appears more complex in rodent cells due to physiological telomerase expression in most somatic normal tissues, great telomere length, and the difficulties of finding suitable in vitro culture conditions. To study in vitro aging of rat lung epithelial cells, we have developed primary culture conditions adapted to rat fresh lung explants and have studied for 1 year (50 passages) the changes in cellular proliferation and mortality, genetic instability, telomerase activity, telomere length, and tumorigenic potential. We have observed an absence of senescence and/or crisis, a transient genetic instability, the persistence of a differentiated Clara cell phenotype, a steady decrease in telomerase activity followed by a low residual activity together with a continuous decrease in telomere length, a constant rate of proliferation, and the acquisition of tumorigenic potential. The bypass of the growth arrest and the acquisition of long-term growth properties could be explained by the loss of p16(INK4a) expression, the ARF/p53 pathway not being altered. In conclusion, these results clearly indicate that, in rat lung epithelial cells, in vitro transformation and acquisition of tumorigenic properties can occur even if the telomere length is still decreasing and telomerase activity remains downregulated. PMID- 12729793 TI - The SHB adapter protein is required for efficient multilineage differentiation of mouse embryonic stem cells. AB - The SH2 domain-containing adapter protein SHB transmits signals from receptor tyrosine kinases regulating diverse processes such as apoptosis and differentiation. To elucidate a role for SHB in cell differentiation, wild-type and R522K (inactive SH2 domain-mutant) SHB were transfected and expressed in mouse embryonic stem (ES) cells. Microarray analysis using Affymetrix U74A chips on undifferentiated ES cells and expression of selected differentiation markers after generation of embryoid bodies were subsequently assessed. Wild-type SHB altered the expression of 16 genes in undifferentiated ES cells, many of which have been found to relate to neural cell function. R522K-SHB altered the expression of 128 genes in undifferentiated ES cells, the majority of which were decreased, including several transcription factors related to development. When grown as embryoid bodies, after 4 days R522K-SHB ES cells were already found to display a different morphological appearance, with an impaired cavity formation that occurred in the absence of altered OCT4 expression. This impairment was reversed by exogenous addition of Matrigel. In addition, R522K-SHB embryoid bodies displayed reduced mRNA contents of the liver protein albumin, the pancreatic proteins amylase, glucagon and insulin after 20 days of differentiation. Matrigel did not restore the impaired expression of albumin in the R522K-SHB cells. Expression of the mesodermal marker cardiac actin and the neural marker neurofilament heavy chain alpha was not affected by wild-type or R522K-SHB overexpression. It is concluded that SHB is required for efficient differentiation of ES cells into embryoid bodies with normal cavities and cells belonging to endodermal lineages. PMID- 12729794 TI - Transmembrane domain of Bcl-2 is required for inhibition of ceramide synthesis, but not cytochrome c release in the pathway of inostamycin-induced apoptosis. AB - Bcl-2 protein plays important roles in the regulation of apoptosis. However, the exact mechanism by which Bcl-2 blocks apoptosis is still unclear. In the present study, we found that overexpression of Bcl-2 in human small cell lung carcinoma Ms-1 cells inhibited not only the release of cytochrome c from mitochondria into cytosol but also de novo ceramide synthesis induced by inostamycin, a phosphatidylinositol turnover inhibitor. To investigate the correlation between the structure of Bcl-2 and its inhibitory function in inostamycin-induced apoptosis, Ms-1 cells that stably overexpress domain-deletional mutants of Bcl-2 were established. Transmembrane domain-deleted Bcl-2 failed to inhibit inostamycin-induced de novo ceramide synthesis, whereas it inhibited inostamycin induced cytochrome c release, indicating that anchoring of Bcl-2 to membrane was a requirement for its inhibitory effect on inostamycin-induced ceramide synthesis, but not cytochrome c release. Thus, the deletion mutant of tarnsmembrane domain of Bcl-2 can suppress inostamycin-induced apoptosis by inhibiting cytochrome c release, a downstream event of ceramide synthesis in the pathway of inostamycin-induced apoptosis. We also found that the BH3 and BH4 domains of Bcl-2 were necessary for inhibition of inostamycin-induced apoptosis, and deletion of BH1 or BH2 did not affect the inhibitory effect of Bcl-2 to inostamycin-induced apoptotic events. PMID- 12729795 TI - Von Willebrand factor targets IL-8 to Weibel-Palade bodies in an endothelial cell line. AB - Vascular endothelial cells are able to store the chemotactic cytokine interleukin 8 (IL-8) in specialized storage vesicles, Weibel-Palade bodies, together with von Willebrand factor (VWF) and P-selectin. We investigated whether VWF plays a role in the sorting of IL-8 into these organelles. We examined the effect of VWF expression on IL-8 targeting in an endothelial cell line (EC-RF24). This cell line has retained the typical phenotypic characteristics of primary endothelial cells but has lost the capacity to produce VWF in appreciable amounts. EC-RF24 cells were retrovirally transduced with a vector encoding a VWF-green fluorescent protein chimera (VWF-GFP). This approach enables direct visualization of the cellular distribution and secretory behavior of the VWF-GFP hybrid. Expression of VWF-GFP resulted in the generation of Weibel-Palade body-like organelles as shown by the colocalization of VWF-GFP and P-selectin. VWF-GFP expressing EC-RF24 cells also showed significant colocalization of VWF-GFP with IL-8 in these storage vesicles. Live cell imaging revealed that the number of VWF-GFP-containing granules decreased upon cell stimulation. These observations indicate that VWF plays an active role in sequestering IL-8 into Weibel-Palade bodies. PMID- 12729796 TI - Effects of expressing lamin A mutant protein causing Emery-Dreifuss muscular dystrophy and familial partial lipodystrophy in HeLa cells. AB - Patients with the autosomal dominant form of Emery-Dreifuss muscular dystrophy (EDMD) or familial partial lipodystrophy (FPLD) have specific mutations in the lamin A gene. Three such point mutations, G465D (FPLD), R482L, (FPLD), or R527P (EDMD), were introduced by site-specific mutagenesis in the C-terminal tail domain of a FLAG-tagged full-length lamin A construct. HeLa cells were transfected with mutant and wild-type constructs. Lamin A accumulated in nuclear aggregates and the number of cells with aggregates increased with time after transfection. At 72 h post transfection 60-80% of cells transfected with the mutant lamin A constructs had aggregates, while only 35% of the cells transfected with wild-type lamin A revealed aggregates. Mutant transfected cells expressed 10 24x, and wild-type transfected cells 20x, the normal levels of lamin A. Lamins C, B1 and B2, Nup153, LAP2, and emerin were recruited into aggregates, resulting in a decrease of these proteins at the nuclear rim. Aggregates were also characterized by electron microscopy and found to be preferentially associated with the inner nuclear membrane. Aggregates from mutant constructs were larger than those formed by the wild-type constructs, both in immunofluorescence and electron microscopy. The combined results suggest that aggregate formation is in part due to overexpression, but that there are also mutant-specific effects. PMID- 12729797 TI - Electrotransfer in differentiated myotubes: a novel, efficient procedure for functional gene transfer. AB - Development of reliable techniques for experimental manipulation of gene expression in multinucleated skeletal muscle fibers is critical for understanding molecular mechanisms involved in both physiology and pathophysiology. At present, viral vectors represent the only method to obtain efficient gene transfer in terminally differentiated myotubes. Here we present an in vitro procedure that relies on the application of a pulsed electric field for transferring naked DNA into differentiated myotubes seeded on coverslips. Compared with standard transfection methods, electroporation was at least 1000 times more efficient, as judged by quantitative determination of luciferase content. Percentage of transfected myotubes averaged around 45%. Moreover, we were successful in transfecting a dominant-negative ADP ribosylation factor 1 (ARF1) mutant, i.e., ARF1N126I, in myotubes, thus interfering with endoplasmic reticulum-Golgi traffic, as indicated by alterations of subcellular distribution of GM130, a cis/medial-Golgi marker. Co-transfection experiments with beta-galactosidase also showed that the ARF1 mutant appeared to inhibit myoblast fusion and could not be used before myotube formation. The present work validates the use of electroporation as a highly efficient approach for gene transfer in fully differentiated myotubes. PMID- 12729798 TI - Overexpressed mortalin (mot-2)/mthsp70/GRP75 and hTERT cooperate to extend the in vitro lifespan of human fibroblasts. AB - The lifespan of human foreskin fibroblasts (HFF5), cultured under standard in vitro conditions (including ambient atmospheric oxygen tension), was extended slightly by expression of exogenous mortalin (mot-2)/mthsp70/Grp75, but not by the catalytic subunit of telomerase, hTERT. Together, mot-2 and hTERT permitted bypass of senescence, a substantial extension of lifespan, and possibly immortalization. This is the first demonstration that mot-2 and telomerase can cooperate in the immortalization process. PMID- 12729799 TI - Multiprotein bridging factor 1 cooperates with c-Jun and is necessary for cardiac hypertrophy in vitro. AB - Cardiac hypertrophy is induced by a number of stimuli and can lead to cardiomyopathy and heart failure. Cardiomyocyte hypertrophy is characterized by increased cell size and altered gene expression. By differential-display polymerase chain reaction and Western blotting we found that the transcriptional coactivator MBF1 was upregulated during hypertrophy in cardiomyocyte cultures. Furthermore, MBF1 protein level increased in two animal models of hypertrophy, angiotensin II treatment and aortic banding. MBF1 antisense oligodeoxynuclotides blocked phenylephrine-induced hypertrophy, suggesting MBF1 plays a key role in hypertrophic growth. In contrast, overexpression of MBF1 potentiated the hormone induced response of the atrial natriuretic peptide promoter. MBF1 overexpressed by transient transfection cooperated with the transcription factor c-Jun in activation of transcription but not with GATA4. MBF1 and c-Jun induced the activity of a transiently transfected atrial natriuretic peptide promoter, whereas neither MBF1 nor c-Jun could induce the promoter alone. Moreover, MBF1 bound to c-Jun in vitro. These data suggest that MBF1 is a transcriptional coactivator of c-Jun regulating hypertrophic gene expression. Inhibitor studies suggested that MBF1 activates the atrial natriuretic peptide promoter independently of the calcineurin and CaMK signaling pathways. Our results indicate that MBF1 participates in hormone-induced cardiomyocyte hypertrophy and activates hypertrophic gene expression as a coactivator of c-Jun. PMID- 12729800 TI - Disintegrin causes proteolysis of beta-catenin and apoptosis of endothelial cells. Involvement of cell-cell and cell-ECM interactions in regulating cell viability. AB - Disintegrins, the snake venom-derived arginine-glycine-aspartic acid-containing peptides, have been demonstrated to inhibit angiogenesis through induction of endothelial cell apoptosis. However, it is not clear how a disintegrin causes endothelial apoptosis. In this study, we elucidated the action mechanism of disintegrin in causing endothelial apoptosis by using rhodostomin as a tool. We showed that cell detachment was observed at the early stage of rhodostomin treatment. It was initiated through the blockade by integrin alphanubeta3 and was accelerated by a mechanical stretch from neighboring cells. Both rhodostomin and poly(HEME) induced a higher percentage of cells at G2-M phase, the cleavage of beta-catenin and poly(ADP-ribose) polymerase during apoptosis, indicating that cell detachment is a prerequisite for rhodostomin-induced apoptosis. Moreover, pp125(FAK) phosphorylation and actin cytoskeleton were affected upon rhodostomin treatment. The activation of caspase-3 but not that of caspase-9 was detected after rhodostomin treatment. In addition, general caspase inhibitors inhibited the cleavage of beta-catenin and poly(ADP-ribose) polymerase, and DNA fragmentation, whereas they did not prevent cell shape change or detachment. According to these results, we concluded that disintegrin-induced endothelial apoptosis is a complex process, not merely caused by a blockade of endothelial integrin alphanubeta3 but also by an accompanied shape change and mechanical stretches among cells. PMID- 12729801 TI - Insulin-like growth factor I-stimulated melanoma cell migration requires phosphoinositide 3-kinase but not extracellular-regulated kinase activation. AB - Dysregulated signaling contributes to altered cellular growth, motility, and survival during cancer progression. We have evaluated the ability of several factors to stimulate migration in WM1341D, a cell line derived from an invasive human vertical growth phase melanoma. Basic fibroblast growth factor, hepatocyte growth factor, interleukin-8, and CCL27 each slightly increased migration. Insulin-like growth factor I (IGF-I), however, stimulated a 15-fold increase in migration. This response required the IGF-I receptor, which activates phosphoinositide 3-kinase (PI3K) and mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathways. Both pathways have been implicated in migration in a variety of cell types, but the signaling required for IGF-I-induced melanoma cell migration is not well defined. IGF-I stimulated activation of MAPK/ERK signaling in WM1341D cells was inhibited by U0126, but a 33-fold higher dose of U0126 was needed to inhibit IGF-I-stimulated cellular migration. In contrast, similar concentrations of either wortmannin or LY294002 were required to inhibit both IGF-I-induced PI3K activation and migration. These results indicate that IGF-I-stimulated migration of WM1341D cells requires PI3K activation but is independent of MAPK/ERK signaling. Determining the contributions of IGF-I signaling pathways to migration will help us to understand melanoma progression and may lead to new therapeutic targets of this highly metastatic cancer. PMID- 12729802 TI - Noncatalytic domain of uPA stimulates human extravillous trophoblast migration by using phospholipase C, phosphatidylinositol 3-kinase and mitogen-activated protein kinase. AB - The serine protease urokinase-type plasminogen activator (uPA) promotes matrix degradation by many cell types, including the invasive extravillous trophoblast (EVT) of the human placenta. The noncatalytic amino-terminal end of uPA binds to uPA receptors (uPARs) expressed by these cells. A highly polarized expression of uPAR-bound uPA at the migration front of EVT cells in situ suggests a functional role of uPA:uPAR interaction in EVT cell migration. The present study examined whether uPA stimulates EVT cell migration, independent of proteolytic function, and investigated some of the signaling pathways involved. Using in vitro propagated EVT cells in Transwell migration assays, both uPA and its noncatalytic amino-terminal fragment (ATF) were shown to stimulate migration through multiporous polycarbonate (pore size 8 microm) membranes. A uPAR-blocking antibody inhibited basal and ATF-stimulated migration. Migration was found to be stimulated by hypoxic conditions, which upregulates uPAR expression; this stimulation was abrogated with the uPAR-blocking antibody, indicating the role of endogenous uPA in EVT cell migration. Spectrofluorometric measurement of cytosolic calcium in cells treated with uPA and ATF demonstrated a rapid rise in [Ca2+](i), which was prevented by pretreatment of cells with thapsigargin, indicating a release from intracellular stores. Both basal and ATF-mediated migratory responses were suppressed in the presence of selective pharmacological inhibitors LY294002, U73122, and U0126, implicating the respective roles of phosphatidinylinositol 3-kinase (PI 3-K), phospholipase C (PLC), and MEK1/2 in basal and ATF-stimulated migratory capacity. Taken together, these results demonstrate that uPA:uPAR interaction stimulates EVT cell migration, independent of uPA enzymatic activity, using the mitogen-activated protein kinase pathway and calcium signaling events including the participation of PI 3-K and PLC. These findings are relevant to clinical conditions of aberrant trophoblast migration, including spontaneous abortion, preeclampsia, and choriocarcinoma. PMID- 12729803 TI - Insulin induces expression of adenosine kinase gene in rat lymphocytes by signaling through the mitogen-activated protein kinase pathway. AB - The activity of adenosine kinase (AK) was significantly impaired in splenocytes isolated from diabetic rats. Administration of insulin to diabetic animals restored AK activity, protein, and mRNA levels in diabetic splenocytes. Experiments performed on cultured rat lymphocytes demonstrated that insulin did not change the stability of AK mRNA. Insulin induced AK gene expression in a dose and time-dependent manner. Maximal increases in AK mRNA (3.9-fold) and activity level (3.7-fold) were observed at the fourth and fifth hours of cell incubation with 10 nM insulin, respectively. The insulin effect on AK expression was not influenced by dibutyryl cAMP (dcAMP). On the other hand dcAMP weakly increased (1.7-fold) basal expression of AK. Exposure of rat lymphocytes to wortmannin, an inhibitor of phosphatidylinositol 3-kinase (PI3K), or rapamycin, an inhibitor of mTOR, did not affect the ability of insulin to stimulate expression of AK. Prior treatment of the cells with 10 microM PD98059, an inhibitor of mitogen-activated protein kinase (MAPK) kinase (MEK) completely blocked insulin-stimulated expression of AK gene. Insulin produced a significant transient increase in the tyrosine phosphorylation of ERK1/2, and PD98059 inhibited this phosphorylation. Furthermore exposure of cells to insulin has resulted in transient phosphorylation of Elk-1 on Ser-383 and sustained elevation of c-Jun and c-Fos protein. The maximal phosphorylation of Elk-1 was observed at 15 min, and was blocked by PD98059. We concluded that insulin stimulates AK gene expression through a series of events occurring sequentially. This includes activation of the MAPK cascade and subsequent phosphorylation of Elk-1 followed by increased expression of c-fos and c-jun genes. PMID- 12729804 TI - Transfer of drug resistance genes in hematopoietic progenitors for chemoprotection: is it still an option? AB - For numerous malignancies a relationship between the intensity of antineoplastic chemotherapy and tumor response has been demonstrated. Myelotoxicity is the main cause of chemotherapy-associated morbidity and of treatment delays. The concept of myeloprotective cytostatic drug resistance gene transfer to normal hematopoietic stem cells (HSC) therefore sparks great enthusiasm. While initial studies using murine retroviral vectors on murine HSC showed that the concept works, a number of clinical studies in the last decade were not informative because of limitations in transduction efficiency and transgene expression.Furthermore, possible side effects such as unforeseen transgene activity and vector integration-based leukemogenesis have been reported. Among others, these developments raised some scepticism against the feasibility of myeloprotective gene transfer. Recently, considerable improvements have been achieved in vector design, HSC manipulation, selection protocols and risk assessment methods which are discussed in detail here. Based on these experimental studies successful clinical trials can now be anticipated. PMID- 12729806 TI - Positive and negative regulation of cellular sensitivity to anti-cancer drugs by FGF-2. AB - The development of resistance to chemotherapy by tumor cells remains a constant limitation to the treatment of cancer. Over the last several years, fibroblast growth factor-2 (FGF-2) has emerged as a growth factor that is capable of modifying the sensitivity of normal and tumor cells to anti-cancer drugs. FGF-2 can produce both drug resistance and drug sensitization in different cell types treated with a variety of cytotoxic agents. An understanding of the differential cellular trafficking and biological activities of the multiple FGF-2 isoforms will help in determining the circumstances under which FGF-2 acts to inhibit versus potentiate drug action. Recent advances suggest that expression of FGF-2 in tumor cells is involved with loss of response to chemotherapy in vivo. Thus, the manipulation of FGF-2 activities to increase the effectiveness of chemotherapeutic agents may have important clinical implications. PMID- 12729805 TI - Pharmacogenomics of ABC transporters and its role in cancer chemotherapy. AB - ATP-binding cassette (ABC) genes play a role in the resistance of malignant cells to anticancer agents. The ABC gene products, including ABCB1 (P-glycoprotein), ABCC1 (MRP1), ABCC2 (MRP2, cMOAT), and ABCG2 (BCRP, MXR, ABCP) are also known to influence oral absorption and disposition of a wide variety of drugs. As a result, the expression levels of these proteins in humans have important consequences for an individual's susceptibility to certain drug-induced side effects, interactions, and treatment efficacy. Naturally occurring variants in ABC transporter genes have been identified that might affect the function and expression of the protein. This review focuses on recent advances in the pharmacogenomics of ABC transporters, and discusses potential implications of genetic variants for the chemotherapeutic treatment of cancer. PMID- 12729807 TI - The interface between ErbB and non-ErbB receptors in tumor invasion: clinical implications and opportunities for target discovery. AB - The molecular switches by which malignant cancer cells evolve from a confined to an invasive state are poorly understood, but seem to involve a progressive activation of a signaling network shared by several growth factor receptors and non-receptor molecules. Abnormal expression of ErbB tyrosine kinase receptors, commonly seen in cancer, is an early event in the invasive process, which makes these receptors exciting targets for drug discovery. The past few years have been full of promise for ErbB targeting in the context of receptor overexpression, but also fraught with disappointment as clinical efficacy has often been hampered by potential problems such as the heterogeneity of receptor expression within the same tumor, and the extensive cooperative signaling among ErbB and non-ErbB receptors. Cooperative signaling is a common characteristic of invasive cancer cells, and is believed to dictate the genetic program that controls invasion switches. Molecular studies on the combinatorial signaling involved in tumor invasion are becoming a fertile area for target discovery in cancer. This review discusses how cooperative signaling between ErbB and non-ErbB receptors regulates tumor invasion and hence provides multiple opportunities for drug discovery, and how current therapies and investigational drugs could pave the way to even more potent alternative combinatorial therapeutic approaches for invasive cancers. PMID- 12729808 TI - Overall injury risk to different drivers: combining exposure, frequency, and severity models. AB - Traffic crash risk assessments should incorporate appropriate exposure data. However, existing US nationwide crash data sets, the NASS General Estimates System (GES) and the Fatality Analysis Reporting System (FARS), do not contain information on driver or vehicle exposure. In order to obtain appropriate exposure data, this work estimates vehicle miles driven (VMD) by different drivers using the Nationwide Personal Transportation Survey (NPTS). These results are combined with annual crash rates and injury severity information from the GES for a comprehensive assessment of overall risk to different drivers across vehicle classes. Data are distinguished by driver age, gender, vehicle type, crash type (rollover versus non-rollover), and injury severity. After correcting for drivers' crash exposure, results indicate that young drivers are far more crash prone than other drivers (per VMD) and that drivers of sports utility vehicles (SUVs) and pickups (PUs) are more likely to be involved in rollover crashes than those driving passenger cars. Although, the results suggest that drivers of SUVs are generally much less crash prone than drivers of passenger cars, the rollover propensity of SUVs and the severity of that crash type offset many of the incident benefits for SUV drivers. PMID- 12729810 TI - Estimating transport fatality risk from past accident data. AB - This paper examines the statistical properties of estimates of fatal accident rates, mean fatalities per accident, and fatality rates when these estimates are based on past accident data. The statistical properties are illustrated by two long-term transport fatal accident datasets from Great Britain, the principal one for railways and the other for roads, chosen to provide a statistical contrast. In both modes, the accident rates have fallen substantially over the long term. Two statistical estimates of current accident and fatality rates are presented for each dataset, one based only on recent data and the other based on estimates of long-term trends. The trend-based estimate is preferred for train accidents because this makes maximum use of the limited and variable data; the recent data are preferred for road accidents because this avoids unnecessary dependence on modelling the trends. For train accidents, the estimated fatality rate based on past accidents is compared with an estimate produced by the railway industry using a risk model. The statistical estimate is less than half the industry's estimate, and the paper concludes that the statistical estimate is to be preferred. PMID- 12729809 TI - The effect of cell phone type on drivers subjective workload during concurrent driving and conversing. AB - The effect of three types of cell phones (hand held, hands free with an external speaker and personal hands free) on total subjective workload (including its constituent components; mental demand, physical demand, temporal demand, performance, effort and frustration) and intelligibility was measured using the NASA-task load index (TLX) and the modified rhyme test (MRT), respectively in 13 experienced drivers (nine male, four female, age range 28-65 years), whilst driving on a rural highway. The drivers rated all components of workload for each type of cell phone to be significantly higher than for a control condition in which no cell phone was used. The mean (standard deviation) total workload was lowest for the personal hands free cell phone (7.1(3.65)) and highest for the hands free speaker phone (10.8 (3.63)) (P<0.001). The mean (standard deviation) intelligibility score was highest for the personal hands free cell phone (74.1 (7.9)) and lowest for the hands free speaker phone (55.0 (10.4)) (P<0.001). Frustration was significantly correlated with total workload (0.60, P<0.001) and intelligibility was significantly correlated with frustration (-0.35, P<0.05). Physical demand was not a high contributor to total workload. It is concluded that a personal hands free cell phone would interfere least with the cognitive demands of driving. PMID- 12729811 TI - Some methodological deficiencies in studies on traffic accident predictors. AB - The paper discusses some methodological problems in (psychological) research on traffic accident predictors and reviews a convenience sample of the literature. Three methodological aspects are identified as being important: reliability of accident predictors, time period for accidents used as dependent variable, and culpability for accidents. Papers are scrutinized and most are found to be wanting in these aspects. Traffic researchers do not adhere to, or hardly even discuss, these basic methodological problems. It is concluded that the current research into (psychological) accident predictors is fraught with methodological deficiencies. Why most studies seem to be deficient in these aspects is not clear, as several researchers have pointed out these problems. PMID- 12729812 TI - An evaluation of four types of railway pedestrian crossing safety intervention. AB - This study evaluated a programme of interventions designed to reduce the incidence of illegal and unsafe crossing of a rail corridor at a city station by boys on their way to and from the adjacent high school in Auckland, New Zealand. The boys were observed crossing before, during, and after implementation of each intervention; in addition, surveys were carried out before and after the programme to discover the boys' attitudes. Rail safety education in school, punishment for every unsafe crossing (continuous punishment), and punishment occasionally for unsafe crossing (intermittent punishment) were associated with significant decreases in unsafe crossing compared with that observed prior to any intervention. General communications about rail safety were not associated with significant decreases in unsafe crossing. When interventions were examined consecutively, unsafe crossing was significantly reduced between the communications and education phases, and even more so between education and continuous punishment, but there was no statistically significant difference in frequency of unsafe crossing between continuous and intermittent punishment. It was concluded that punishment may be more effective in reducing unsafe behaviour in this type of situation than targeted education, and is much more effective than communications to heighten awareness. PMID- 12729813 TI - Effect of cellular telephone conversations and other potential interference on reaction time in a braking response. AB - This experiment studied the effect of phone conversations and other potential interference on reaction time (RT) in a braking response. Using a laboratory station which simulated the foot activity in driving, 22 research participants were requested to release the accelerator pedal and depress the brake pedal as quickly as possible following the activation of a red brake lamp. Mean reaction time was determined for five conditions: (a) control, (b) listening to a radio, (c) conversing with a passenger, (d) conversing using a hand-held phone, and (e) conversing using a hands-free phone. Results indicated that conversation, whether conducted in-person or via a cellular phone caused RT to slow, whereas listening to music on the radio did not. PMID- 12729814 TI - The distraction effects of phone use during a crucial driving maneuver. AB - Forty-two licensed drivers were tested in an experiment that required them to respond to an in-vehicle phone at the same time that they were faced with making a crucial stopping decision. Using test track facilities, we also examined the influence of driver gender and driver age on these dual-task response capacities. Each driver was given task practice and then performed a first block of 24 trials, where one trial represented one circuit of the test track. Half of the trials were control conditions in which neither the stop-light was activated nor was the in-vehicle phone triggered. Four trials required only stop-light response and a further four, phone response only. The remaining four trials required the driver to complete each task simultaneously. The order of presentation of specific trials was randomized and the whole sequence was repeated in a second block giving 48 trials per driver. In-vehicle phone response also contained an embedded memory task that was evaluated at the end of each trial circuit. Results confirmed our previous observation that in the dual-task condition there was a slower response to the light change. To compensate for this slowed response, drivers subsequently brake more intensely. Most importantly, we recorded a critical 15% increase in non-response to the stop-light in the presence of the phone distraction task which equates with increased stop-light violations on the open road. These response patterns varied by driver age and driver gender. In particular, age had a large effect on task components that required speed of response to multiple, simultaneous demands. Since driving represents a highly complex and interactive environment, it is not possible to specify a simplistic relationship between these distraction effects and outcome crash patterns. However, we can conclude that such in-vehicle technologies erode performance safety margin and distract drivers from their critical primary task of vehicle control. As such it can be anticipated that a causal relation exists to collision events. This is a crucial concern for all in-vehicle device designers and for the many safety researchers and professionals seeking to reduce the adverse impacts of vehicle collisions. PMID- 12729815 TI - Effect of lowering the alcohol limit in Denmark. AB - On 1 March 1998, the Danish per se limit was lowered from 0.08 to 0.05% blood alcohol concentration (BAC) for motor vehicle drivers. Based on accident data and drivers' drinking habits before and after the amendment, the effect of the new limit has been evaluated. Interviews revealed a significant decrease in the number of drinks that drivers allow themselves to drink within a 2-h period before driving. The proportion of drivers, who would not drink at all or only have one drink, increased from 71% before the amendment to 80% after the amendment. Drivers with changed drinking habits most often stated the lower limit as the main reason for having less alcohol. However, based on accident data from the first year after the amendment, this has not resulted in a marked decrease in the proportion of injury accidents with impaired motor vehicle drivers (BAC>/=0.05%) compared to all injury accidents. On the contrary, the proportion of fatal accidents with drink-drivers compared to all fatal accidents has increased in the after-period. The total number of drink-driving sentences were a little larger in 1999 than in 1997 because of the lower limit, but a significant change from higher towards lower alcohol levels can be seen. PMID- 12729816 TI - The benefits of improved car secondary safety. AB - The term 'secondary safety' refers to the protection that a vehicle provides its occupants when involved in an accident. This paper studies information from the British database of road accident reports between 1980 and 1998, to estimate the reduction in the number of occupant casualties over these years which may be attributed to improvements to secondary safety in cars. The paper shows that the proportion of driver casualties who are killed or seriously injured (KSI) is lower for modern cars than for older cars. The reduction of this proportion is used to assess the improvement in secondary safety. Statistical models are developed to represent the proportion with 'year of first registration' as one of the independent variables, although only an incomplete assessment of the benefits of improved secondary safety can be made with the available data. The assessment compares the number of casualties that would have been expected if secondary safety had remained at the level found in cars first registered in 1980 with the actual casualty numbers. It is estimated that improved secondary safety reduced the number of drivers KSI by at least 19.7% in 1998, in comparison with what might have occurred if all cars had had that lower level of secondary safety. This figure relates to all cars on the road in 1998, and rises to 33%, when confined to the most modern cars (those which were first registered in 1998). PMID- 12729817 TI - Comparison of road crashes incidence and severity between some French counties. AB - Our aim is to compare traffic safety among several counties in France, and explore whether observed differences can be explained by differences in road types distribution and by differences in socio-economic characteristics between counties. Traffic safety is measured by incidence and severity, where incidence is defined by the ratio of counts of injury accidents and exposure, measured by the amount of kilometres driven. Severity is measured by the ratio between fatal and injury accidents. These indexes are analysed in the framework of Generalised Linear Models: counts of injury accidents are analysed with a Negative Binomial regression, which accounts for over-dispersion. Severity being the proportion of fatal accidents among injury accidents corresponds to the probability of a Binomial setting and this is modelled by a logistic regression. This modelling provides an easy way to adjust for covariates such as road type, environment (urban/rural) and evolution over time, and to test their possible interactions. We find that the time trend of each indice (incidence and severity) is the same across counties and across road types. There is a significant interaction between county and road type, meaning that, first, differences in traffic safety between counties are not fully explained by different road type distributions, and second, that the "ranking" of counties in term of incidence or severity varies according to the road type considered, and vice-versa. It was planned to explore global characteristics of the counties (driving and socio-economic data) as possible explanatory factors of differences between counties, but the existence of an interaction of county with road types shows the necessity of collecting and exploring characteristics of the sub-levels of road type within county. PMID- 12729818 TI - Getting off your bike: cycling accidents in Great Britain in 1990-1999. AB - The paper extracts and tabulates selected incidence and fatality rates of cycling accidents recorded by the police in Great Britain during 1990-1999, in a database of over 30,000 standardised reports of fatal or serious injury accidents. Since usable estimates of exposure to risk in cycling are not available, the concept of exposure invariance is developed to estimate the relative risk of different sorts of bicycle/vehicle encounter. Any bias from under-reporting of serious injury accidents is shown to be boundable if the probability of under-reporting is constant. PMID- 12729819 TI - How would setting policy priorities according to cost-benefit analyses affect the provision of road safety? AB - This paper analyses how setting priorities for road safety strictly according to cost-benefit analysis would affect the provision of road safety in Norway and Sweden. The paper is based on recent analyses of the efficiency of road safety policies in these two countries. The argument sometimes made by critics of cost benefit analysis, that only a few road safety measures are cost-effective (have benefits greater than costs), is not supported. Cost-effective road safety policies could prevent between 50 and 60% of the current number of road accident fatalities in both Norway and Sweden, if pursued consistently during a period of 10 years (2002-2011). If current policies are continued, only about 10-15% of the current number of road accident fatalities are likely to be prevented during the next 10 years. A number of sources of inefficiency in road safety policy are identified. A source of inefficiency is anything that prevents policy priorities from being set according to cost-benefit analysis. These include: (a) lack of power, which means that national governments do not have the formal authority to introduce a certain road safety measure, in Europe, this applies to new vehicle safety standards, which are passed almost exclusively by the European Union; (b) the existence of social dilemmas, which means that measures that are cost effective from a societal point of view are not so from the point of view of individual road users; (c) priority given to other policy objectives, in particular regional development. Scarcity of resources, which obtains when public budgets have to be increased to make room for all cost-effective measures, was not found to be a constraint. All cost-effective measures can be funded within current budgets, provided the use of inefficient measures ceases. PMID- 12729820 TI - Analysis of injuries among pilots involved in fatal general aviation airplane accidents. AB - The purpose of this study was to analyze patterns of injuries sustained by pilots involved in fatal general aviation (GA) airplane accidents. Detailed information on the pattern and nature of injuries was retrieved from the Federal Aviation Administration's autopsy database for pilots involved in fatal GA airplane accidents from 1996 to 1999. A review of 559 autopsies revealed that blunt trauma was the primary cause of death in 86.0% (N=481) of the autopsies. The most commonly occurring bony injuries were fracture of the ribs (72.3%), skull (55.1%), facial bones (49.4%), tibia (37.9%) and pelvis (36.0%). Common organ injuries included laceration of the liver (48.1%), lung (37.6%) heart (35.6%), and spleen (30.1%), and hemorrhage of the brain (33.3%) and lung (32.9%). A fractured larynx was observed in 14.7% of the cases, a finding that has not been reported in literature until now. It was observed that individuals who sustained brain hemorrhage were also more likely to have fractures of the facial bones rather than skull fractures. PMID- 12729821 TI - Assessing the effectiveness of minimum legal drinking age and zero tolerance laws in the United States. AB - The objective of this research was to determine the extent to which the decline in alcohol-related highway deaths among drivers younger than age 21 years can be attributed to raising the minimum legal drinking age (MLDA) and establishing zero tolerance (0.02% blood alcohol concentration (BAC) limit for drivers younger than age 21 years) laws. Data on all drivers younger than age 21 years involved in fatalities in the United States from 1982 to 1997 were used in the study. Quarterly ratios of BAC-positive to BAC-negative drivers in each of the 50 states where analyzed in a pooled cross-sectional time-series analysis. After accounting for differences among the 50 states in various background factors, changes in economic and demographic factors within states over time, and the effects of other related laws, results indicated substantial reductions in alcohol-positive involvement in fatal crashes were associated with the two youth-specific laws. The policy of limiting youth access to alcohol through MLDA laws and reinforcing this action by making it illegal for underage drivers to have any alcohol in their system appears to have been effective in reducing the proportion of fatal crashes involving drinking drivers. PMID- 12729822 TI - A behavioral comparison between motorcyclists and a matched group of non motorcycling car drivers: factors influencing accident risk. AB - Motorcyclists and a matched group of non-motorcycling car drivers were assessed on behavioral measures known to relate to accident involvement. Using a range of laboratory measures, we found that motorcyclists chose faster speeds than the car drivers, overtook more, and pulled into smaller gaps in traffic, though they did not travel any closer to the vehicle in front. The speed and following distance findings were replicated by two further studies involving unobtrusive roadside observation. We suggest that the increased risk-taking behavior of motorcyclists was only likely to account for a small proportion of the difference in accident risk between motorcyclists and car drivers. A second group of motorcyclists was asked to complete the simulator tests as if driving a car. They did not differ from the non-motorcycling car drivers on the risk-taking measures but were better at hazard perception. There were also no differences for sensation seeking, mild social deviance, and attitudes to riding/driving, indicating that the risk-taking tendencies of motorcyclists did not transfer beyond motorcycling, while their hazard perception skill did. PMID- 12729823 TI - Traffic fatalities and injuries: the effect of changes in infrastructure and other trends. AB - An analysis of how various road infrastructure improvements affect traffic related fatalities and injuries is conducted while controlling for other factors known to affect overall safety. The road infrastructure elements analysed include total lane miles, the fraction of lane miles in different road categories (interstates, arterial, and collector roads), the average number of lanes for each road category, and lane widths for arterials and collector roads. Other variables that are controlled for in the study include total population, population age cohorts, per capita income, per capita alcohol consumption, seat belt legislation (and seat-belt usage), and a proxy variable that represents underlying changes in medical technology. The data used is a cross-sectional time series database of US states and is analysed using a fixed effects negative binomial regression that accounts for heterogeneity in the data. Data from all 50 states over 14 years is used. Results strongly refute the hypothesis that infrastructure improvements have been effective at reducing total fatalities and injuries. While controlling for other effects, it is found that demographic changes in age cohorts, increased seat-belt use, reduced alcohol consumption and increases in medical technology have accounted for a large share of overall reductions in fatalities. PMID- 12729824 TI - Sleep debt, sleepiness and accidents among males in the general population and male professional drivers. AB - Men from the general population and male professional lorry and bus drivers were surveyed with regard to sleep habits and motor vehicle and other types of accidents. A random sample of 4000 men in the general population of Dalarna County in mid-Sweden were mailed a questionnaire and served as referents. A total of 1389 male professional lorry and bus drivers from this county responded to the same questionnaire. A total of 161 of the drivers also underwent a sleep study in their homes. The proportion of total accidents was higher among the professional drivers as compared with the males in the population, P=0.03. Reports on traffic accidents were the same in both groups, but the professional drivers reported more accidents at leisure compared with referents, P<0.0001. Accidents of any kind, traffic accidents included, among those affected by both snoring and apneas, were not reported more in either of the groups. At the sleep study, 17% of those examined received the diagnosis of obstructive sleep-apnea syndrome (OSAS). The professional drivers reported proportionally more sleep debt than the referents, P<0.001. Among referents, traffic accidents at leisure, traffic accidents while commuting and accidents at work increased in proportion to sleep debt (P<0.001, 0.006 and 0.002, respectively). The finding that self-perceived sleep debt may have an adverse effect on males in the general population and male professional drivers concerning accident likelihood should have an impact on prevention. These results stress the need to educate the general population on the importance of complying with our biological need of sleep. PMID- 12729826 TI - A bivariate zero-inflated Poisson regression model to analyze occupational injuries. AB - The aim of many occupational safety interventions is to reduce the incidence of injury. However, when measuring intervention effectiveness within a period, population-based accident count data typically contain a large proportion of zero observations (no injury). This situation is compounded where injuries are categorized in a binary manner according to an outcome of interest. The distribution thus comprises a point mass at zero mixed with a non-degenerate parametric component, such as the bivariate Poisson. In this paper, a bivariate zero-inflated Poisson (BZIP) regression model is proposed to evaluate a participatory ergonomics team intervention conducted within the cleaning services department of a public teaching hospital. The findings highlight that the BZIP distribution provided a satisfactory fit to the data, and that the intervention was associated with a significant reduction in overall injury incidence and the mean number of musculoskeletal (MLTI) injuries, while the decline in injuries of a non-musculoskeletal (NMLTI) nature was marginal. In general, the method can be applied to assess the effectiveness of intervention trials on other populations at high risk of occupational injury. PMID- 12729825 TI - Superstition, risk-taking and risk perception of accidents among South African taxi drivers. AB - The aim of this study was to investigate taxi drivers' superstition and risk perception of accidents as well as risk-taking in an urban area in South Africa. One hundred and thirty drivers of minibuses, so-called "taxis" were interviewed on the basis of: (1) a superstition scale; (2) a risk-taking scale; (3) a list of perceived causes of road traffic accidents. Drivers showed largely superstitious attitudes and expressed a high degree of risk-taking behavior. Superstition was positively correlated with the number of self-reported accidents the drivers had been involved in and the number of accidents they had witnessed. Path analysis revealed a direct path from superstition to accident involvement while the influence of formal education was negligible. Risk-taking was inversely correlated with driving experience and the number of accidents witnessed but not so with the number of accidents involved in. There was no clear pattern of associations between superstition and risk-taking and perceived causes of accidents. Superstition and risk-taking were slightly and inversely correlated with each other. It is concluded that superstition represents an attitude that is associated with a driver's accident risk, and further research on superstitious attitudes among South African drivers is advocated. PMID- 12729827 TI - Square wave voltammetric determination of the angiotensin-converting enzyme inhibitors cilazapril, quinapril and ramipril in pharmaceutical formulations. AB - The angiotensin-converting enzyme inhibitors cilazapril, quinapril and ramipril are reduced at a hanging mercury drop electrode in the pH range 3.5-13 using Britton-Robinson buffers as supporting electrolyte and KCl as ionic medium. Square wave voltammetry has proved to be the most suitable electroanalytical technique for the quantitative voltammetric determination of these antihypertensive drugs. Optimisation of the chemical and instrumental variables was carried out. Analyses were performed in 0.02 M borate buffer at pH 9.5 and 0.5 M KCl as ionic medium, using a pulse amplitude of 50 mV and a frequency of 150 Hz. A linear relationship between peak current and concentration was found in the interval 0.5-8 microg/ml for cilazapril and up to 6 microg/ml for quinapril and ramipril, allowing the direct determination of their pharmaceutical formulations alone or mixed with hydrochlorothiazide. Good accuracy and repeatativity were obtained. PMID- 12729828 TI - Synthesis and cytotoxic activity of 1,3-benzodioxole derivatives. Note II. AB - A series of 1,3-benzodioxoles (2-12) were synthesized and evaluated for their in vitro ability to inhibit the growth of three human tumor cell lines. No cytotoxic effects were noticed with any of the test compounds at a concentration of 10(-4) M. PMID- 12729829 TI - 3D QSAR studies of the interaction between beta-tubulin and microtubule stabilizing antimitotic agents (MSAA). A combined pharmacophore generation and pseudoreceptor modeling approach applied to taxanes and epothilones. AB - Based on the conformer of paclitaxel extracted from the experimental tubulin structure, a pharmacophoric model has been generated and used to find the chemical features common to the taxane and epothilone classes of compounds. This original alignment has been translated into the experimental tubulin binding site obtaining an assembly subsequently submitted to the pseudoreceptor modeling approach. As a result, an original 3D QSAR model, able to evaluate, at a quantitative level, the relationships between the molecular structures and biological data of the studied compounds, has been obtained. PMID- 12729831 TI - Tumor necrosis factor-alpha production-regulating activity of phthalimide derivatives in genetically modified murine melanoma cells B78H1. AB - The effect of imides, monothioimides, trimellitimides, as well as 5'-deoxy-5' phthaloylamino-derivatives of azidothymidine on tumor necrosis factor-alpha (TNF alpha) production by genetically modified murine B78H1 melanoma cells transduced with the gene for human TNF-alpha (B78/TNF) was investigated. It was found that N (adamant-1-yl)monothiophthalimide (1e) and N-(adamant-2-yl)-monothiophthalimide (1f) showed over 200% enhancing of TNF-alpha production while some of imides were inhibitors. PMID- 12729830 TI - An improved method for preparation of cefpodoxime proxetil. AB - Cefpodoxime proxetil, a third-generation cephalosporin for oral administration, was synthesized by a method based on the following sequence of reactions: acylation of 7-aminocephalosporanic acid (7-ACA) with S-benzothiazol-2-yl(2-amino 4-thiazolyl)(methoxyimino)thioacetate (MAEM), chloroacetylation of the cefotaxime formed with chloroacetyl chloride, esterification of the acid function with 1 iodoethyl isopropyl carbonate and final cleavage of chloroacetamide protective group by treatment with thiourea in N,N-dimethylacetamide. The developed procedure allows us to obtain better yields of cefpodoxime proxetil and to eliminate the final purification step by column chromatography, necessary during the synthesis of this antibiotic by the previously reported methods. PMID- 12729832 TI - Determination of two ternary mixtures containing phenobarbitone by second derivative of the ratio spectrum-zero-crossing and HPLC methods. AB - A spectrophotometric method is developed for the determination of ternary mixtures with overlapping spectra. The method is based on the use of the second derivative of the ratio spectrum with a zero-crossing technique. The ratio spectrum was obtained by dividing the absorption spectrum of the mixture by that of one of the components. The concentration of the other components are then determined from their respective calibration graphs treated similarly. The method is accurate, non-destructive and do not require resolutions of equations. The method has been applied for the resolution of two ternary mixtures, namely, phenobarbitone, methylphenobarbitone and phenytoin (1), and phenobarbitone, papaverine HCl and piperazine acefyllinate (2). Also, a HPLC method was developed for determination of phenobarbitone, papaverine and HCL and piperazine acefyllinate. The HPLC method depends upon using ODS column with a mobile phase consisting of acetonitrile-5 mM aqueous heptane sulfonic acid sodium salt (50:50, v/v) and adjusted to apparent pH 4 using acetic acid. Quantitation was achieved with UV detection at 220 nm based on peak area. The proposed methods were applied for the determination of the two ternary combinations in synthetic mixtures and in commercial pharmaceutical products. The results obtained were precise and accurate. PMID- 12729833 TI - An in vitro investigation for vaginal bioadhesive formulations: bioadhesive properties and swelling states of polymer mixtures. AB - Bioadhesive tablet formulations have been developed for mucosal application. Sixteen different bioadhesive tablet formulations were prepared and evaluated. Their bioadhesion to vaginal mucosa were studied by tensile testing method. The swelling behaviour of the tablets in three different solutions was also investigated. In addition, the effect of the formulations on pH of the medium was followed. The most favorable formulation resulted a mixture of Carbopol 934 and Pectin (2:1). The highest bioadhesive strength, the highest swelling volume and the lowest pH reduction were obtained with this formulation. PMID- 12729834 TI - Investigations on mefenamic acid sustained release tablets with water-insoluble gel. AB - Mefenamic acid (MA) has analgesic, anti-inflammatory and antipyretic properties. Available conventional dosage forms are capsules and film-coated tablets. No commercial sustained release preparation of MA exists in the market. The usual oral dose is 250 or 500 mg and reported half-life is 2 h. Sodium alginate (NaAL) is the sodium salt of alginic acid, a natural polysaccharide extracted from marine brown algae. It has the ability to form a water-insoluble gel with a bivalent metal ions as calcium. Therefore, NaAL has been studied for preparing sustained release formulations in pharmaceutical technology. In this study, tablet formulations containing different ratios of NaAL and calcium gluconate (CaGL) were prepared by direct compression method. In vitro release studies were carried out using USP 23 basket method and release data were kinetically evaluated. According to release studies, it can be emphasized that NaAL and CaGL can be used for design of sustained release preparation of MA. PMID- 12729835 TI - In vitro cytotoxicity to human cells in culture of some phenolics from olive oil. AB - The neutral red in vitro cytotoxicity assay was used to evaluate the comparative responses of human cells isolated from tissues of the oral cavity to olive oil phenolics. The cell lines used included normal gingival fibroblasts, immortalized, nontumorigenic gingival epithelial cells, and carcinoma cells from the salivary gland. No differences in the relative sensitivities to the phenolics amongst the three cell types were noted. In general, for all cell types, the sequence of increasing cytotoxicity was: oleuropein aglycone>oleuropein glycoside, caffeic acid>o-coumaric acid>cinnamic acid>>tyrosol, syringic acid, protocatechuic acid, vanillic acid. Cytotoxicity was noted only at phenolic concentrations far exceeding those attainable after habitual consumption, thus indicating that consumption of phenol-rich olive oil is safe. PMID- 12729836 TI - 6beta-acetoxy nortropane regulated processing of amyloid precursor protein in CHOm1 cells and rat brain. AB - The effects of the muscarinic receptor agonist 6beta-acetoxy nortropane on amyloid precursor protein (APP) processing were studied in both transfected Chinese hamster ovary cells stably expressing muscarinic M(1) receptors (denoted as CHOm(1) cell line) and in cerebral cortical and hippocampal slices. Exposure of CHOm(1) cells to 6beta-acetoxy nortropane for 1 h significantly increased the secretion of secretory amyloid precursor protein (derived from alpha-secretase cleavage) in a concentration-dependent manner. In the same system, 6beta-acetoxy nortropane reduced the beta-amyloid peptide production. Similar results were obtained in hippocampal and cerebral cortical slices, with 6beta-acetoxy nortropane administration resulting in an increase in secretory amyloid precursor protein and a decrease in beta-amyloid peptide release. The increase of secretory amyloid precursor protein secretion was abolished by preincubation with selective muscarinic M(1) receptor antagonist pirenzepine, but not by preincubation with selective muscarinic M(2) receptor antagonist methoctramine, suggesting that 6beta-acetoxy nortropane promotes secretory amyloid precursor protein release in the brain via muscarinic M(1) receptor activation. These results suggest that 6beta-acetoxy nortropane could exert a beneficial effect on the progress of Alzheimer's disease by promoting amyloid precursor protein processing through alpha-secretase. PMID- 12729838 TI - Loss of anti-cataleptic effect of scopolamine in mice lacking muscarinic acetylcholine receptor subtype 4. AB - Motor dysfunction associated with dyskinesia can be caused by imbalance between dopaminergic and cholinergic actions. Antimuscarinic agents are used to treat extrapyramidal symptoms in Parkinson's disease and extrapyramidal side effects of antipsychotics. These therapeutic effects are mediated by blockade of the striatal muscarinic receptors, which comprise five distinct subtypes (M(1-5)). To evaluate the role of muscarinic M(4) receptors, we have generated mutant mice lacking this subtype (muscarinic M(4) receptor-knockout mice) and analyzed their cataleptic responses induced by haloperidol (an animal model of extrapyramidal side effects). While the muscarinic M(4) receptor-knockout mice developed the cataleptic response normally, systemic administration of scopolamine could not suppress the cataleptic response. These results suggest that acute, but not chronic, blockade of muscarinic M(4) receptors plays important roles in the therapeutic effects of antimuscarinic agents. PMID- 12729837 TI - P2Y receptor specific for diadenosine tetraphosphate in lung: selective inhibition by suramin, PPADS, Ip5I, and not by MRS-2197. AB - Extracellular dinucleotides, which act as signaling molecules in a variety of systems, may regulate fluid homeostasis in the human lung by activation of a specific P2Y receptor subtype. Previously, we presented evidence for a G protein coupled P2Y receptor with high affinity for dinucleotides in both rat and human lung tissue. In a human bronchial epithelial cell line (HBE-1), diadenosine polyphosphates (Ap(n)A, n=2-6) increase intracellular Ca(2+). The aim of the present work was to find additional evidence that, in these cells, the receptors selectively activated by diadenosine polyphosphates are distinct from already known P2Y receptors, which are activated by the mononucleotides ATP or UTP. We tested antagonists suitable to classify P2Y receptor subtypes. The P2Y(1) receptor-selective antagonist 2'-deoxy-N(6)-methyl adenosine 3',5'-diphosphate (MRS-2197) did not affect Ca(2+) mobilization induced by diadenosine tetraphosphate (Ap(4)A). However, suramin, pyridoxal phosphate-6-azo(benzene-2,4 disulfonic acid) (PPADS) and diinosine pentaphosphate (Ip(5)I) inhibited the Ca(2+) response by 96%, 92% and 32%, respectively. Moreover, these results were confirmed by assessing the specific binding of [3H]Ap(4)A to membranes from human and rat lung. Suramin (100 microM), PPADS (400 microM) and Ip(5)I (200 microM), reduced [3H]Ap(4)A binding in lung membrane preparations by 66%, 77% and 80%, respectively. The Ap(4)A-induced Ca(2+) response in HBE-1 cells was inhibited to a much greater extent by these antagonists than the ATP- or UTP-evoked Ca(2+) rise. Thus, Ap(4)A in lung epithelial cells also activates a still unidentified P2Y receptor that is specific for dinucleotides over mononucleotides. PMID- 12729840 TI - Ramatroban, a TP receptor antagonist, improves vascular responses to acetylcholine in hypercholesterolemic rabbits in vivo. AB - Recent studies show that 8-iso-prostaglandin F(2alpha), a member of F(2) isoprostane family, acts as a vasoconstrictor via TP receptor activation; and its local release may contribute to an abnormal vasomotor tone associated with hypercholesterolemia. The purpose of this study was to examine whether ramatroban, a TP receptor antagonist, improves abnormal vascular reactivity in vivo in hypercholesterolemic rabbits. The plasma 8-iso-prostaglandin F(2alpha) levels in hypercholesterolemic groups were significantly higher than those in normal groups. The treatment by ramatroban reversed the attenuation of the vascular response to acetylcholine in hypercholesterolemic groups. However, L N(G)-nitroarginine methyl ester, a nitric oxide synthase inhibitor, did not inhibit the protective effects of ramatroban. Attenuation of the vascular response to acetylcholine in hypercholesterolemic rabbits was significantly enhanced by 8-iso-prostaglandin F(2alpha). Attenuation of the vascular response to acetylcholine by a cholesterol-rich diet and 8-iso-prostaglandin F(2alpha) was canceled by ramatroban. These findings suggest that ramatroban improves the vascular response in vivo to acetylcholine in hypercholesterolemic rabbits by blocking the action of 8-iso-prostaglandin F(2alpha). PMID- 12729839 TI - Taurine reduces ammonia- and N-methyl-D-aspartate-induced accumulation of cyclic GMP and hydroxyl radicals in microdialysates of the rat striatum. AB - Acute ammonia neurotoxicity caused by intraperitoneal administration of ammonium salts is mediated by overactivation of N-methyl-D-aspartate (NMDA) receptors, with ensuing generation of free radicals and extracellular accumulation of cyclic GMP (cGMP) arising from stimulation of nitric oxide (NO) synthesis. In this study, infusion of ammonium chloride or NMDA into the striata of rats via microdialysis probes increased the contents of cyclic GMP and hydroxyl radicals in the microdialysates. Co-infusion of taurine virtually abolished both the ammonia- and NMDA-induced accumulation of cGMP. Taurine also attenuated accumulation of hydroxyl radicals evoked by either treatment. This result is the first evidence of a potential of taurine to attenuate the effects of NMDA receptor overactivation by ammonia in vivo and points to the inhibition of the NMDA receptor-mediated NO synthesis as a possible mechanism of its neuroprotective action. Taurine or its blood-brain barrier penetrating analogues may be applicable in treatment of ammonia-induced neurological deficits. PMID- 12729841 TI - Induction of vasorelaxation through activation of nitric oxide synthase in endothelial cells by brazilin. AB - The vasorelaxant activity of Caesalpinia sappan L., a traditional Chinese medicine, and its major component brazilin were investigated in isolated rat aorta and human umbilical vein endothelial cells. In isolated rat aorta, C. sappan L. extract and brazilin relaxed phenylephrine-induced vasocontraction and increased cyclic guanosine 3',5'-monophosphate (cGMP) content. Induction of vasorelaxation of brazilin was endothelium-dependent and could be markedly blocked by pretreatment with nitric oxide synthase (NOS) inhibitor, N(G)-nitro-L arginine methyl ester (L-NAME); N(G)-monomethyl-L-arginine acetate (L-NMMA) and guanylyl cyclase inhibitor, methylene blue; 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin 1-one (ODQ) and nitric oxide (NO) scavenger, hemoglobin. The increasing cGMP content induced by brazilin was also blocked by pretreatment with L-NAME, methylene blue, and the removal of extracellular Ca(2+). In human umbilical vein endothelial cells, brazilin dose-dependently induced an increase in NO formation and NOS activity, which were greatly attenuated by either the removal of extracellular Ca(2+) or the chelating of intracellular Ca(2+) chelator, 1,2-bis(2 aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA-AM). Moreover, brazilin dose-dependently induced the influx of extracellular Ca(2+) in human umbilical vein endothelial cells. Collectively, these results suggest that brazilin induces vasorelaxation by the increasing intracellular Ca(2+) concentration in endothelial cells of blood vessels and hence activating Ca(2+)/calmodulin dependent NO synthesis. The NO is released and then transferred into smooth muscle cells to activate guanylyl cyclase and increase cGMP content, resulting in vasorelaxation. PMID- 12729842 TI - Structural and functional characterization of gastric mucosa and central nervous system in histamine H2 receptor-null mice. AB - To examine the physiological role of the histamine H(2) receptor, histamine H(2) receptor-null mice were generated by homologous recombination. Histamine H(2) receptor-null mice, which developed normally and were fertile and healthy into adulthood, exhibited markedly enlarged stomachs and marked hypergastrinemia. The former was due to hyperplasia of gastric gland cells (small-sized parietal cells, enterochromaffin-like cells and mucous neck cells which were rich in mucin), but not of gastric surface mucous cells, which were not increased in number as compared with those in wild-type mice despite the marked hypergastrinemia. Basal gastric pH was slightly but significantly higher in histamine H(2) receptor-null mice. Although carbachol but not gastrin induced in vivo gastric acid production in histamine H(2) receptor-null mice, gastric pH was elevated by both muscarinic M(3) and gastrin antagonists. Thus, both gastrin and muscarinic receptors appear to be directly involved in maintaining gastric pH in histamine H(2) receptor-null mice. Interestingly, gastric glands from wild-type mice treated with an extremely high dose of subcutaneous lansoprazole (10 mg/kg body weight) for 3 months were very similar to those from histamine H(2) receptor-null mice. Except for hyperplasia of gastric surface mucous cells, the findings for gastric glands from lansoprazole-treated wild-type mice were almost identical to those from gastric glands from histamine H(2) receptor-null mice. Therefore, it is possible that the abnormal gastric glands in histamine H(2) receptor-null mice are secondary to the severe impairment of gastric acid production, induced by the histamine H(2) receptor disruption causing marked hypergastrinemia. Analyses of the central nervous system (CNS) of histamine H(2) receptor-null mice revealed these mice to be different from wild-type mice in terms of spontaneous locomotor activity and higher thresholds for electrically induced convulsions. Taken together, these results suggest that (1) gastrin receptors are functional in parietal cells in histamine H(2) receptor-null mice, (2) abnormal gastric glands in histamine H(2) receptor-null mice may be secondary to severe impairment of gastric acid production and secretion and (3) histamine H(2) receptors are functional in the central nervous system. PMID- 12729843 TI - Dexamethasone reduces lung eosinophilia, and VCAM-1 and ICAM-1 expression induced by Sephadex beads in rats. AB - Airway eosinophilia is one of the key pathophysiologic features in asthma. The endothelial adhesion molecules, vascular cell adhesion molecule (VCAM-1) and intercellular adhesion molecule (ICAM-1), have previously been shown to play a crucial role in eosinophil recruitment into the inflamed airway. We have investigated the effects of dexamethasone on eosinophilia into the bronchoalveolar lavage fluid, and the upregulation of VCAM-1 and ICAM-1 expression, measured by immunoblotting, induced by i.v. injection of Sephadex beads into rats. The beads significantly increased the lung eosinophilia, and expression of VCAM-1 and ICAM-1 in the lung. Pretreatment with dexamethasone (0.1 to 2 mg/kg i.p.) strongly inhibited all the airway inflammatory events in a dose dependent manner. In conclusion, glucocorticoids may be potent inhibitors of lung eosinophilia, at least in part, due to the prevention of the upregulation of VCAM 1 and ICAM-1 expression. PMID- 12729845 TI - Introduction. PMID- 12729844 TI - Anti-inflammatory activity of erycristagallin, a pterocarpene from Erythrina mildbraedii. AB - Erycristagallin, a pterocarpene isolated from Erythrina mildbraedii, was tested in vitro for its antioxidant properties on the stable 2,2-diphenyl-1-pycryl hydrazyl (DPPH) free radical and on the arachidonic acid metabolism. In addition, erycristagallin was tested on different experimental models of inflammation, such as the acute and chronic inflammation induced by the application of 12-O tetradecanoylphorbol 13-acetate (TPA) on mice and the phospholipase A(2)-induced mouse paw oedema test. In the carrageenan-induced mouse paw oedema test, the ethyl acetate extract obtained from E. mildbraedii showed anti-inflammatory activity, and erycristagallin was isolated as the active principle. In vivo, erycristagallin significantly inhibited the phospholipase A(2)-induced mouse paw oedema as well as the mouse ear oedema induced by TPA (ID(50)<10 microg/ear). Moreover, it significantly reduced the chronic inflammation and leukocyte infiltration induced by repeated application of TPA. In vitro, erycristagallin inhibited the arachidonic acid metabolism via the 5-lipoxygenase pathway in rat polymorphonuclear leukocytes (IC(50)=23.4 microM), but had no effect on cyclooxygenase-1 metabolism in human platelets, while showing antioxidant activity in the DPPH test. As with other phenolics, the anti-inflammatory activity of erycristagallin may be based on its capacity to inhibit the arachidonic acid metabolism via the 5-lipoxygenase pathway. PMID- 12729846 TI - Prevention and management of chronic heart failure in management of asymptomatic patients. AB - Symptomatic heart failure is preceded by a somewhat prolonged asymptomatic stage in many patients. The number of patients with asymptomatic heart dysfunction is about 4-fold greater than the number of patients with clinically overt heart failure. Pharmacologic treatment with angiotensin-converting enzyme inhibitors and beta-blockers (in particular carvedilol) of asymptomatic patients with systolic left ventricular (LV) dysfunction can prevent or delay the occurrence of symptoms and reduce mortality in the long term. Thus, it would be of utmost importance to recognize and appropriately treat these patients before they develop heart failure symptoms. The cost-effectiveness of screening for asymptomatic heart dysfunction in the general population and in cohorts at risk has not been extensively evaluated. A normal electrocardiogram has a high negative predictive value in patients at risk. Echocardiography is the best tool for diagnosis and characterization of heart dysfunction, but extensive use is limited by availability and cost. Natriuretic peptides (brain natriuretic peptide and N-terminal pro-brain natriuretic peptide) are very sensitive markers of heart dysfunction and volume overload, and their measurement has been proposed as a first-line test to select patients who need echocardiography. The definition of the etiology of LV dysfunction--in particular, of the ischemic etiology--has prognostic and therapeutic implications. In addition to revascularization, pharmacologic treatment with antiplatelets and statins is helpful in preventing new ischemic events and the development of heart failure. The prevention, or at least the delay, of clinical manifestations of heart failure is strongly related to an effective approach to the asymptomatic stage. Therefore, it is important to educate the entire medical community, particularly physicians in the primary care setting, about recognition and treatment of these patients. PMID- 12729847 TI - Prevention and management of chronic heart failure in patients at risk. AB - The prevalence and incidence of chronic heart failure (HF) have now reached epidemic proportions. However, the issue of the prevention of HF has been raised only recently. New US guidelines have introduced a new classification system that includes 4 categories: patients at risk, patients with asymptomatic left ventricular dysfunction, patients with symptomatic HF, and those with refractory HF. Because coronary artery disease is the major cause of HF, its risk factors are also those of HF. Hypertension favors the development of HF through accelerated atherosclerosis and increased left ventricular wall stress and hypertrophy. Left ventricular hypertrophy is also a powerful risk factor for HF, independent of blood pressure. Angiotensin-converting enzyme (ACE) inhibitors, beta-blockers, and diuretics are the antihypertensive agents that have been associated with favorable effects in patients with overt HF. Therefore, they may be preferred in the prevention of this syndrome. Diabetes is the most frequent noncardiac comorbidity of HF and is independently associated with an increased risk. Normalization of glycemic and glycosylated hemoglobin levels is a desirable goal of treatment. However, no direct evidence exists in the prevention of HF. A greater control of the other risk factors (eg, hypertension, hyperlipidemia) is, on the other hand, particularly important. Beta-blockers and ACE inhibitors have both been shown to have favorable effects across all spectrums of severity of HF. The ACE inhibitor ramipril has also been shown to prevent the development of HF in patients at risk without left ventricular dysfunction. The role of antiplatelet agents, warfarin, and statins is clear in the prevention of the coronary artery disease. However, it has not been adequately assessed in patients with HF and awaits the results of ongoing trials. PMID- 12729848 TI - Pharmacologic therapy for patients with chronic heart failure and reduced systolic function: review of trials and practical considerations. AB - Heart failure (HF) is a complex clinical syndrome resulting from any structural or functional cardiac disorder impairing the ability of the ventricles to fill with or eject blood. The approach to pharmacologic treatment has become a combined preventive and symptomatic management strategy. Ideally, treatment should be initiated in patients at risk, preventing disease progression. In patients who have progressed to symptomatic left ventricular dysfunction, certain therapies have been demonstrated to improve survival, decrease hospitalizations, and reduce symptoms. The mainstay therapies are angiotensin-converting enzyme (ACE) inhibitors and beta-blockers (bisoprolol, carvedilol, and metoprolol XL/CR), with diuretics to control fluid balance. In patients who cannot tolerate ACE inhibitors because of angioedema or severe cough, valsartan can be substituted. Valsartan should not be added in patients already taking an ACE inhibitor and a beta-blocker. Spironolactone is recommended in patients who have New York Heart Association (NYHA) class III to IV symptoms despite maximal therapies with ACE inhibitors, beta-blockers, diuretics, and digoxin. Low-dose digoxin, yielding a serum concentration <1 ng/mL can be added to improve symptoms and, possibly, mortality. The combination of hydralazine and isosorbide dinitrate might be useful in patients (especially in African Americans) who cannot tolerate ACE inhibitors or valsartan because of hypotension or renal dysfunction. Calcium antagonists, with the exception of amlodipine, oral or intravenous inotropes, and vasodilators, should be avoided in HF with reduced systolic function. Amiodarone should be used only if patients have a history of sudden death, or a history of ventricular fibrillation or sustained ventricular tachycardia, and should be used in conjunction with an implantable defibrillator [corrected]. Finally, anticoagulation is recommended only in patients who have concomitant atrial fibrillation or a previous history of cerebral or systemic emboli. PMID- 12729849 TI - Nonpharmacologic care of heart failure: counseling, dietary restriction, rehabilitation, treatment of sleep apnea, and ultrafiltration. AB - The prognosis of patients with chronic congestive heart failure (CHF) depends not only on pharmacologic therapy but also on nonpharmacologic aspects. A complete and ongoing education program for treating CHF includes an understanding of the causes of CHF, symptoms, diet, salt and fluid restriction, drug regimen, compliance, physical and work activities, lifestyle changes, and measures of self control. Moreover, the nonpharmacologic treatment (dietary modifications, lifestyle, physical exercise, and health care education) must be inserted in a multidisciplinary program organized by the physician in conjunction with the health system, the nurses, and, especially, the patients themselves, who must understand their disease and the many therapeutic options. Cardiologists should treat patients in a clear and comprehensible way, and other specialists (dietitians, physiotherapists, psychologists, nurses, and social workers), together with the patient's family, should strive for the best living conditions for the patient. In this way, the treatment of CHF can improve the quantity and quality of life and save a significant amount in health care costs. PMID- 12729850 TI - Nonpharmacologic care of heart failure: patient, family, and hospital organization. AB - Patients with severe heart failure require large quantities of health care resources, and more intensive interventions are not always related to a decrease in need for medical care, including hospitalization. A thoughtful approach to the efficient and expeditious allocation of these resources is required. In fact, the nonpharmacologic therapy of patients with chronic congestive heart failure (CHF) has to include the complete explanation of various topics, such as causes of heart failure, symptoms, diet, salt and fluid restriction, drug regimen, compliance, physical and work activities, lifestyle changes, and measures of self control. This article describes how the relationship among patient, family, friends, and different heart-care workers (physicians, nurses, psychologists, rehabilitation technicians, dietitians, hospital personnel) can greatly affect therapeutic efficacy. PMID- 12729852 TI - Prevention and management of chronic heart failure with electrical therapy. AB - Sudden cardiac death is responsible for >40% of patients with heart failure losing their lives. Thus, the prevention of life-threatening cardiac arrhythmias is a major goal in the management of heart failure. In several randomized clinical trials, electrical therapy with the implantable cardioverter defibrillator (ICD) has proved superior to medical antiarrhythmic therapy in both the secondary and primary prevention of sudden cardiac death in patients with reduced left ventricular function. In addition to the severity of left ventricular dysfunction, the etiology of the cardiomyopathy appears to be a determinant in the benefit derived from this form of electrical therapy. Whereas patients with ischemic cardiomyopathy clearly show improved survival with ICD therapy, outcome data in patients with nonischemic cardiomyopathy are less convincing. The major challenge lies in the risk stratification of patients with heart failure for arrhythmic death. Catheter ablation is another form of electrical therapy that can help in the treatment of patients with heart failure. In patients with a tachycardia-mediated cardiomyopathy because of drug-refractory atrial fibrillation with rapid ventricular response, catheter ablation of the atrioventricular node and pacemaker implantation can effectively restore a physiologic heart rate, often with dramatic regression of left ventricular dysfunction. In patients with frequent ICD therapies because of frequent recurrences of ventricular tachyarrhythmias, catheter ablation of ventricular tachycardia can be an effective adjunctive therapy. New catheter ablation techniques and new atrial pacing algorithms can also significantly reduce the atrial fibrillation burden in patients with heart failure who are particularly susceptible to decompensation because of atrial fibrillation. Pacing for hemodynamic benefit in heart failure has evolved from dual-chamber pacing modes with optimized atrioventricular delay to biventricular pacing resulting in cardiac resynchronization. This new treatment modality for advanced heart failure has been shown to result in significant symptomatic and hemodynamic improvement. PMID- 12729851 TI - Biventricular pacing in heart failure: back to basics in the pathophysiology of left bundle branch block to reduce the number of nonresponders. AB - Cardiac resynchronization therapy is a novel nonpharmacologic approach to treating patients who have advanced heart failure with left bundle branch block (LBBB). Such a therapy is based on the original theory that synchronous biventricular pacing is able to reduce the interventricular delay caused by LBBB in patients with heart failure. Although there is convincing evidence that biventricular pacing increases the left ventricular ejection fraction, decreases mitral regurgitation, and improves symptoms caused by heart failure, the percentage of nonresponders to such therapy has been described as high as about one third of patients with heart failure having LBBB. Factors responsible for this relatively high prevalence are reviewed, the most important of them probably being left intraventricular dyssynchrony, which can persist after biventricular pacing, notwithstanding right and left interventricular resynchronization. Such a dyssynchrony, as evaluated by tissue Doppler imaging, may be because of the discordance between the site of the left ventricular pacing and the site of the left ventricular delay. Therefore, to characterize the pathophysiologic pattern of LBBB, the investigators suggest an assessment of the electromechanical dysfunction with a noninvasive reliable technique, such as tissue Doppler imaging, which can be repeated after biventricular pacing. PMID- 12729853 TI - Combining electrical therapies for advanced heart failure: the Milan experience with biventricular pacing-defibrillation backup combination for primary prevention of sudden cardiac death. AB - Biventricular pacing (BVP) improves hemodynamics and symptoms in patients with heart failure with bundle branch block. Patients with a left ventricular ejection fraction <0.35 and ventricular tachyarrhythmias are at risk of sudden cardiac death, and they benefit most from implantable cardioverter defibrillators (ICDs). No study has evaluated the efficacy of the BVP-ICD combination in patients with heart failure with no history of syncope or sustained ventricular tachycardia (VT) or ventricular fibrillation (VF). Our prospective, observational study was performed on 135 consecutive patients with heart failure (aged, 64 +/- 11 years; 76% male; New York Heart Association functional class, 3.1 +/- 0.8; ejection fraction 0.28 +/- 0.06; ischemic heart failure, 43%; QRS interval duration, 153 +/- 11 msec) treated at our cardiac pacing unit between January 1999 and April 2001. In the first year (control phase), BVP alone was implanted. After that, BVP with ICD backup was used (prophylactic phase). Follow-up data were obtained by outpatient visits with electrocardiographic and echocardiographic examinations done at 3-month intervals. For patients who died, we examined hospital records, death certificates, and autopsy reports. Follow-up time averaged 840 days. The first 47 patients received BVP alone. During follow-up study, 19% of these patients died suddenly, and 11% died of worsening heart failure. None of the patients who died suddenly had hemodynamic deterioration or BVP malfunction before the event. The BVP-ICD group comprised 88 patients (18% with VT/VF inducibility on electrophysiologic testing). During follow-up study, 32% of these patients (18% with positive electrophysiologic testing) had VT/VF episodes successfully treated by ICD; 5% received inappropriate discharges on atrial fibrillation; and 6% died of heart failure with 1 sudden cardiac death. Cox proportional hazards model in the BVP-ICD group compared with BVP alone revealed hazard ratios for all-cause mortality and sudden cardiac death of 0.76 (95% confidence interval [CI], 0.58 to 0.96; p = 0.01) and 0.08 (95% CI, 0.05 to 0.42; p <0.01), respectively, adjusting for baseline characteristics and follow-up duration. Mortality in patients with heart failure remains high after BVP implantation, mainly because of sudden cardiac death. Although there are limitations with an observational study, our experience suggests that ICD backup grants increased security in BVP patients without conventional class I ICD indications. PMID- 12729854 TI - Surgical methods to reverse left ventricular remodeling in congestive heart failure. AB - In this review article, we describe the most common surgical procedures currently used to reverse or arrest remodeling of the left ventricle in patients with congestive heart failure (CHF). The selection of the appropriate operation in a patient is a complex decision-making process, rigorously based on pathophysiologic considerations. In this population, all factors affecting the surgical risk should be carefully evaluated preoperatively, and surgery should be recommended when definite benefits in survival and quality of life can be reasonably predicted. Quite often, patients with CHF require a combination of different procedures to address all the pathophysiologic components determining the clinical picture. In particular, in this review we describe the surgical restoration of the left ventricle, the isolated coronary artery bypass graft procedure, the correction of mitral regurgitation, diastolic support (from dynamic cardiomyoplasty to passive containment), and mechanical assist devices. Moreover, in the future, the role of surgery in the treatment of CHF will be strongly modified by the advent of gene therapy, cell therapy, and engineered artificial myocardial tissue. PMID- 12729855 TI - Surgical therapy in advanced heart failure. AB - Congestive heart failure (CHF) affects about 1% of adults in the United States and is a contributing factor in >250,000 deaths per year. In an increasingly elderly population, the surgical treatment of CHF made great progress during the past 3 decades, consuming enormous health care resources. Heart transplantation is still the most effective therapy for end-stage heart disease, with the 10-year survival rate after transplantation approaching 50%. Efforts to increase the supply of donor organs have failed to improve the shortage, underscoring the crucial need for alternatives to cardiac allotransplantation. Alternative surgical options to end-stage heart transplantation are rapidly evolving. Left ventricular assist devices have been used as a bridge to heart transplantation for patients who otherwise might die awaiting a new heart. There is also continued interest in the use of these devices either to bridge patients to full recovery or to destination therapy, without the need for heart replacement. Left ventricular reconstruction, including the Batista and Dor procedures, along with mitral valve repair, cardiomyoplasty, and extreme coronary artery bypass graft surgery, are now being increasingly performed as alternative options. The history, status, and personal experience of surgical treatment of end-stage heart disease are discussed. PMID- 12729856 TI - Early microdose drug studies in human volunteers can minimise animal testing: Proceedings of a workshop organised by Volunteers in Research and Testing. AB - Testing the safety and efficacy of a successful human medicine involves many laboratory animals, which can sometimes be subjected to considerable suffering and distress. Also, it is necessary to extrapolate from the test species to humans. UK and European legislation requires that Replacement, Reduction and Refinement of animal procedures (the Three Rs) are implemented wherever possible. Over the last decade, there has been substantial progress with applying in vitro and in silico methods to both drug efficacy and safety testing. This paper is a report of the discussions and recommendations arising from a workshop on the role that might be played by human volunteer studies in the very early stages of drug development. The workshop was organised in November, 2001 by Volunteers in Research and Testing, a group of individuals in the UK which launched an initiative in 1994 to identify where and how human volunteers can participate safely in biomedical studies to replace laboratory animals. It was considered that conducting pre-Phase I very low dose human studies (sub-toxic and below the dose threshold for measurable pharmacological or clinical activity) could enable drug candidates to be assessed earlier for in vivo human pharmacokinetics and metabolism. Moreover, accelerator mass spectrometry (AMS), nuclear magnetic resonance (NMR) spectroscopy and positron emission tomography (PET) are potentially useful spectrometric and imaging methods that can be used in conjunction with such human studies. Some, limited animal tests would still be required before pre-Phase I microdose studies, to take account of the potential risk posed by completely novel chemicals. The workshop recommended that very early volunteer studies using microdoses should be introduced into the drug development process in a way that does not compromise volunteer safety or the scientific quality of the resulting safety data. This should improve the selection of drug candidates and also reduce the likelihood of later candidate failure, by providing in vivo human ADME data, especially for pharmacokinetics and metabolism, at an earlier stage in drug development than is currently the case. PMID- 12729857 TI - Functional role of adenosine receptor subtypes in the regulation of blood-brain barrier permeability: possible implications for the design of synthetic adenosine derivatives. AB - The objective of this investigation was to determine the functional role of adenosine receptor subtypes in the regulation of blood-brain barrier (BBB) permeability. The presence of the equilibrative es and ei nucleoside transporters at the BBB was also determined. Studies were conducted in an experimental in vitro BBB model comprising bovine brain capillary endothelial cells (BCECs) and rat astrocytes (RAs). The presence of the receptors and transporters was investigated by a combination of RT-PCR and radioligand binding assays. Changes in paracellular permeability were investigated on basis of changes in trans endothelial-electrical-resistance (TEER) and transport of paracellular markers. In BCECs the presence of A(2A) and A(3) receptors and the es nucleoside transporter was demonstrated. The A(1) receptor was absent, while the presence of the A(2B) receptor and the ei nucleoside transporter remained uncertain. In RAs the presence of all four receptor subtypes and the es and ei nucleoside transporters was demonstrated. Upon application of selective agonists no significant changes in TEER or the transport of the paracellular markers were observed. The functional role of adenosine receptor subtypes in regulating the paracellular permeability of the BBB is probably small. It is unlikely therefore that the BBB transport of synthetic adenosine analogues is modified by permeability changes. The es nucleoside transporter might play a role in the BBB transport of synthetic adenosine analogues. PMID- 12729858 TI - Cellular uptake and metabolism of clodronate and its derivatives in Caco-2 cells: a possible correlation with bisphosphonate-induced gastrointestinal side-effects. AB - PURPOSE: To investigate possible reasons for the low frequency of GI side-effects of clodronate, even though clodronate is known to be metabolised into a cytotoxic nucleotide analogue (AppCCl(2)p) by many cell types. The effects of some lipophilic prodrugs of clodronate were also studied. METHODS: The effects of clodronate and its lipophilic derivatives on the proliferation and viability of Caco-2 cells were examined using an MTT assay. The intracellular uptake of 14C clodronate and the accumulation of a clodronate metabolite (AppCCl(2)p) in Caco-2 cells were evaluated using ion-pairing HPLC-ESI-MS. RESULTS: Clodronate had little effect on growth of proliferating, or the viability of confluent, Caco-2 cells. The uptake of clodronate by Caco-2 cells was only about 0.04% of total clodronate. The potentially cytotoxic clodronate metabolite, AppCCl(2)p, was detected in Caco-2 cell extracts after 3 h of exposure. Dianhydride- and triPOM clodronate were metabolised to AppCCl(2)p more efficiently and also affected the viability of Caco-2 cells more than clodronate. CONCLUSIONS: Clodronate appears to be metabolised into a cytotoxic ATP-analogue (AppCCl(2)p) by any cell type capable of internalising the drug. However, the cytotoxicity depends on the degree of uptake of clodronate. Due to the very low initial uptake of clodronate by epithelial Caco-2 cells, they do not accumulate sufficient intracellular concentrations of AppCCl(2)p to affect cell function. This explains the low frequency of gastrointestinal side-effects caused by oral clodronate therapy. PMID- 12729859 TI - Stability of sufentanil and levobupivacaine solutions and a mixture in a 0.9% sodium chloride infusion stored in polypropylene syringes. AB - We have evaluated the chemical and microbiological stability of sufentanil citrate, levobupivacaine hydrochloride and a mixture in a 0.9% sodium chloride infusion in order to provide background information on the storage of a sufentanil-levobupivacaine mixture in polypropylene (PP) syringes. Chemical assays were performed by HPLC on days 0, 1, 2, 3, 8, 14, 23, 28 and 30 after storage at 4, 21, and 36 degrees C. Microbiological stability was evaluated under aseptic conditions using a laminar air flow station, with a grade A environment and a B background. The samples taken for microbiological analysis were collected immediately after preparation of the solutions and then after 7, 14, 21 and 28 days storage. At 4 degrees C the sufentanil citrate solution was stable for 23 days. At 21 degrees C the sufentanil citrate solution maintained chemical stability for 3 days, but thereafter the concentration of sufentanil decreased 15% from day 3 to day 8. At 36 degrees C a similar decrease was noticed from day 1 to day 3. On the contrary, the levobupivacaine hydrochloride solution maintained chemical stability for 28 days at 4 and 21 degrees C and for 23 days at 36 degrees C. The sufentanil-levobupivacaine mixture maintained chemical stability for 28 days at 4, 21 and 36 degrees C. The sufentanil and levobupivacaine solutions and the mixture studied maintained microbiological stability for 28 days. According to the chemical and microbiological stability studies, the sufentanil-levobupivacaine mixture in PP syringes could be stored for 28 days at 4 and 21 degrees C. PMID- 12729861 TI - Pharmacodynamic considerations in bioequivalence assessment: comparison of novel and existing metrics. AB - This study addresses the utility of pharmacodynamic considerations to the assessment of bioequivalence (BE) studies. A novel methodology was developed and the performance of classic, nonclassic and novel BE indices was evaluated using extensive simulations of BE trials generated from a classic pharmacokinetic (PK)/pharmacodynamic (PD) model. Three novel indices based on drug's pharmacodynamics were developed and served as criteria for the assessment of all BE indices. Modified power curves were constructed and used for the analysis of BE trials from a PD point of view. All BE indices of either purely PK or PD nature were classified in a semiquantitative manner according to their strictness in declaring BE. The partial area until the peak concentration followed by the two newly proposed metrics (MARD, MARD(w1)) exhibited the most strict performance in declaring BE irrespective of the PK scenarios examined. The study opens new avenues in BE assessment since it places more emphasis on the PD aspects of the formulations. PMID- 12729860 TI - Anandamide prodrugs. 1. Water-soluble phosphate esters of arachidonylethanolamide and R-methanandamide. AB - Phosphate esters of arachidonylethanolamide (AEA) and R-methanandamide were synthesized and evaluated as water-soluble prodrugs. Various physicochemical properties (pK(a), partition coefficient, aqueous solubility) were determined for the synthesized phosphate esters. The chemical stability of phosphate esters was determined at pH 7.4. In vitro enzymatic hydrolysis rates were determined in 10% liver homogenate, and in a pure enzyme-containing (alkaline phosphatase) solution at pH 7.4. The intraocular pressure (IOP) lowering properties of R-methanandamide phosphate ester were tested on normotensive rabbits. The phosphate promoiety increased the aqueous solubility of the parent compounds by more than 16500-fold at pH 7.4. Phosphate esters were stable in buffer solutions, but rapidly hydrolyzed to their parent compounds in alkaline phosphatase solution (t(1/2)<<15 s) and liver homogenate (t(1/2)=8-9 min). The phosphate ester of R-methanandamide reduced IOP in rabbits. These results indicate that the phosphate esters of AEA and R-methanandamide are useful water-soluble prodrugs. PMID- 12729862 TI - Enantioselective transport and CYP3A4-mediated metabolism of R/S-verapamil in Caco-2 cell monolayers. AB - We have evaluated the passive and carrier-mediated intestinal transport and CYP3A4-mediated metabolism of R/S-verapamil with respect to dose dependency and enantioselectivity in modified Caco-2 cells. The present in vitro results were compared to published data from human in vivo and rat in situ jejunal perfusions with R/S-verapamil. Caco-2 cell permeability to enantiomers of verapamil and norverapamil was weakly concentration dependent (2.5-100 microM). While Caco-2 permeability to verapamil was 2.6- to 3.7-fold lower than in the human jejunum, it was 1.4- to 2.3-fold higher than in rats. However, all three models classified R- and S-verapamil as high permeability compounds according to the biopharmaceutical classification system. In accordance with human and rat data, R/S-verapamil was transported to a minor extent by carrier-mediated mechanisms in Caco-2 cells. Neither the passive nor the carrier-mediated permeability was enantioselective in any of the three models. CYP3A4-mediated demethylation to R/S norverapamil was enantioselective in Caco-2 cells. Apparent V(max) and K(m) values for the conversion of R-verapamil were 3.2 pmol/min/insert and 0.7 microM, respectively, and for S-verapamil, 5.4 pmol/min/insert and 0.6 microM, respectively. The enantioselectivity in the CYP3A4-metabolism observed in Caco-2 cells was in agreement with human data, but not with rat data, indicating that Caco-2 cells better reflect the human small intestine in this regard. However, all three models suggested that intestinal permeability to verapamil is unaffected by CYP3A4-activity. In summary, modified Caco-2 cells and human jejunum were qualitatively related with respect to R-and S-verapamil transport and CYP3A4-metabolism. PMID- 12729863 TI - Development and characterization of naproxen-chitosan solid systems with improved drug dissolution properties. AB - The solubilizing and amorphizing properties toward naproxen (a poorly water soluble antiinflammatory drug) of chitosan, an emerging pharmaceutical biopolymer, have been investigated. Solid binary systems at different drug/polymer ratios have been prepared according to different techniques (mixing, cogrinding, kneading, coevaporation) using chitosan at low (CS-L(w)) and medium (CS-M(w)) molecular weight, and tested for dissolution properties. Drug-carrier interactions were investigated in both the liquid and solid state, by phase solubility analysis, differential scanning calorimetry, X-ray powder diffractometry, FT-IR spectroscopy, and scanning electron microscopy. Drug dissolution parameters improved with increasing the polymer amount in the mixture, reaching the highest values at the 1:9 (w/w) drug/polymer ratio, and CS L(w) was more efficacious than CS-M(w). Cogrinding was the most effective technique, showing the strongest amorphizing effect toward the drug and enabling an increase of more than ten times its relative dissolution rate. Coground mixtures at 3:7 (w/w) drug/polymer ratio were able to give directly compressed tablets which maintained unchanged the improved drug dissolution properties. Enhancer dissolution properties combined with its direct compression feasibility and antiulcerogenic action make CS-L(w) an optimal carrier for developing fast release oral solid dosage forms of naproxen. PMID- 12729864 TI - Aripiprazole in the treatment of schizophrenia: safety and tolerability in short term, placebo-controlled trials. AB - Aripiprazole is a novel antipsychotic with a unique mechanism of action. Presented here is a pooled analysis of safety and tolerability data from all completed short-term, placebo-controlled trials in schizophrenia from the aripiprazole clinical development program. Data were analyzed from five 4- to 6 week double-blind multicenter studies of patients hospitalized with acute relapse of schizophrenia or schizoaffective disorder randomized to aripiprazole (n=932), placebo (n=416), or haloperidol (n=201). Daily aripiprazole doses ranged from 2 to 30 mg. Safety assessments included adverse event (AE) reports, EPS scales, ECGs, weight, and prolactin, glucose and cholesterol levels. Aripiprazole was well tolerated, with similar AE incidence rates to placebo, and lower rates than haloperidol for akathisia, extrapyramidal syndrome and somnolence. Objective EPS assessments demonstrated no significant differences between aripiprazole and placebo on Simpson-Angus Scale (SAS) scores, no dose-dependent effects on Barnes Akathisia scores, and significant reductions in Abnormal Involuntary Movement Scale (AIMS) scores from baseline vs. placebo (p1 microM. The proapoptotic effect of TBTC on fish blood cells, and involvement of mitochondrial pathways was also investigated by verifying the release of cytochrome c, activation of caspase-3 and the presence of "DNA laddering". Although mitochondrial activity was much more strongly affected in erythrocytes, leukocytes incubated in the presence of TBTC showed the characteristic features of apoptosis after only 1 h of incubation. Longer exposures, up to 12 h, were required to trigger an apoptotic response in erythrocytes. PMID- 12729920 TI - 17beta-estradiol inhibits NADPH oxidase activity through the regulation of p47phox mRNA and protein expression in THP-1 cells. AB - In this report, we demonstrate that NADPH oxidase is activated by tumor necrosis factor-alpha (TNF-alpha) plus interferon-gamma (IFN-gamma) in human monocytic cells (THP-1 cells) differentiated with phorbol ester (PMA) and that physiological concentration of 17beta-estradiol inhibits NADPH oxidase activity in THP-1 cells stimulated with TNF-alpha plus IFN-gamma. This effect is mediated by estrogen receptor based on estrogen receptor antagonist (ICI 182, 780) that diminishes inhibition by 17beta-estradiol. This inhibition is specific in 17beta estradiol because 17alpha-estradiol, testosterone and progesterone do not inhibit NADPH oxidase activity. Activation of NADPH oxidase induced by TNF-alpha plus IFN gamma is caused by up-regulation of p47(phox) (cytosolic component of NADPH oxidase) expression. 17beta-Estradiol prevents the up-regulation of p47(phox) mRNA and protein expression. This prevention of p47(phox) expression depends on the inhibition of NF-kappaB activation. Our results implicate that 17beta estradiol has an anti-atherosclerotic effects through the improvement of nitric oxide (NO) bioavailability caused by the regulation of superoxide (O(2)(-)) production. PMID- 12729921 TI - Human serum albumin and its structural variants mediate cholesterol efflux from cultured endothelial cells. AB - In the present study, we used the human EA.hy926 endothelial cell line as the model system to investigate the effect of human serum albumin (HSA) and its structural variants on cholesterol efflux. Initial studies showed that HSA promoted cholesterol efflux in a dose- and time-dependent manner, reaching a plateau at 10 mg/ml at 90 min. As a control, gelatin displayed no significant effect on efflux, while HSA was significantly more efficient than ovalbumin and bovine serum albumin (BSA) in promoting cholesterol efflux. Equal molar concentrations of HSA and apolipoprotein A-I (apoA-I) showed that apoA-I had considerably higher efficiency in efflux. However, the prevailing high plasma concentrations of HSA may compensate for its lower efflux rate compared to apoA I. To characterize the mechanism of HSA-mediated cholesterol efflux, we studied the effects of cAMP and temperature on efflux using both EA.hy926 endothelial cells and murine RAW 264.7 macrophages. We found that HSA-mediated efflux occurred via a cAMP-independent and relatively temperature-insensitive pathway. We next examined the nature of HSA-cholesterol interaction by comparing the effects of various HSA mutants to wild-type HSA on cholesterol efflux. We found specific interactions between subdomains 2A and 3A and cholesterol, as indicated by the changes in the efflux rate of various HSA mutants. In conclusion, our study provides evidence for the role of HSA in cholesterol efflux, and shows that the substitution of specific amino acid residues in subdomains of 2A and 3A may be important structural determinants in its ability to bind to cholesterol and participate in cholesterol efflux. PMID- 12729922 TI - Basic peptide system for efficient delivery of foreign genes. AB - Certain peptides containing high percentage of cationic amino acids are known to efficiently translocate through the cell membrane. This principle was previously exploited for delivery of variety proteins. We had observed that various basic peptides of earlier studies, though not specifically use for gene delivery, contain DNA or RNA binding domains. In the present study, we reported on arginine peptides, which form DNA complexes that efficiently transfect various cell lines. The transfection abilities of the peptides were observed by green fluorescent protein (GFP) and beta-galactosidase gene expression in 293T, HeLa, Jurkat, and COS-7 cells. We found superior transfection activity of arginine peptides compared with commercially available efficient transfection agents. The expression of marker genes induced by arginine peptides was partially inhibited in the presence of heparan sulfate, chondroitin sulfate B and C, or both heparinase III and chondroitinase ABC. The transfection proficiency of these peptides was affected by endosomotropic reagent as well as low temperature (4 degrees C). Finally, we have investigated the potential of arginine peptides as a delivery agent for gene therapy, by attempting to deliver herpes simplex virus thymidine kinase (HSV-TK) gene into tumor cells. HSV-TK transfected tumor cells exhibited sensitivity to the antiviral drug ganciclovir (GCV), leading to cell death. Taken together, these data demonstrate that arginine peptide is proficient for transfection, indicating its potentially benefit to studies in gene therapy and gene delivery in a range of model organisms. PMID- 12729924 TI - Cellular localisation and nuclear export of the human bZIP transcription factor TCF11. AB - TCF11 is a ubiquitous transcription factor of the CNC-bZIP family. The activity of this vital protein is strictly regulated and we have previously published that the two major translated protein forms show a clearly different transactivation ability in transient transfections. Only the full-length form is active in a variety of mammalian cells [J. Biol. Chem. 276 (2001) 17641]. Here we further investigate the complex regulation of TCF11, studying the cellular localisation of some of the different protein isoforms. The full-length form is located both in the cytoplasm and the nucleus, while the internally initiated shorter protein form is restricted to nuclear localisation. A nuclear export signal (NES) localised in the N-terminus of TCF11 is responsible for the active nuclear export of the protein. This export is highly sensitive to leptomycin B (LMB) and is largely blocked by mutating three of the leucine residues in the signal region. These results indicate that export occurs through the Crm1-mediated pathway. Due to alternative splicing within the tcf11 gene, different isoforms of the longer protein form are produced. Some of these isoforms, one identical to Nrf1, lack the NES and are thereby restricted to nuclear localisation. PMID- 12729923 TI - Effects of beta-adrenergic agonists on bone-resorbing activity in human osteoclast-like cells. AB - In the present study, we demonstrate for the first time that beta-adrenergic agonists stimulate bone-resorbing activity in human osteoclast-like multinucleated cells (MNCs). Osteoclast-like MNCs constitutively expressed mRNA for alpha1B-, alpha2B- and beta2-adrenergic receptor (AR) in addition to characteristic markers of mature osteoclast, such as calcitonin receptor (CT-R), tartrate-resistant acid phosphatase (TRAP), alphaV-chain of integrin (Int alphaV), carbonic anhydrase II (CA-II) and cathepsin K (Cathe K). Epinephrine (1 microM; alpha,beta-adrenergic agonist) up-regulated expression of Int alphaV, CA II and Cathe K in the osteoclast-like MNCs. Osteoclastic resorbing activity was markedly increased by isoprenaline (1 microM; beta-adrenergic agonist), moderately by epinephrine, but poorly by phenylephrine (1 microM; alpha1 adrenergic agonist). The actin ring, which was suggested to be correlated with bone-resorbing activity, was clearly observed in osteoclast-like MNCs treated with isoprenaline and epinephrine, but faintly in those treated with phenylephrine. These findings suggest that beta-adrenergic agonists directly stimulate bone-resorbing activity in matured osteoclasts. PMID- 12729925 TI - Recycling of vitamin C from its oxidized forms by human endothelial cells. AB - Endothelial cells encounter oxidant stress due to their location in the vascular wall, and because they generate reactive nitrogen species. Because ascorbic acid is likely involved in the antioxidant defenses of these cells, we studied the mechanisms by which cultures of EA.hy926 endothelial cells recycle the vitamin from its oxidized forms. Cell lysates reduced the ascorbate free radical (AFR) by both NADH- and NADPH-dependent mechanisms. Most NADH-dependent AFR reduction occurred in the particulate fraction of the cells. NADPH-dependent reduction resembled that due to NADH in having a high affinity for the AFR, but was mediated largely by thioredoxin reductase. Reduction of dehydroascorbic acid (DHA) required GSH and was both direct and enzyme dependent. The latter was saturable, half-maximal at 100 microM DHA, and comparable to rates of AFR reduction. Loading cells to ascorbate concentrations of 0.3-1.6 mM generated intracellular DHA concentrations of 20-30 microM, indicative of oxidant stress in culture. Whereas high-affinity AFR reduction is the initial and likely the preferred mechanism of ascorbate recycling, any DHA that accumulates during oxidant stress will be reduced by GSH-dependent mechanisms. PMID- 12729926 TI - Signaling through Cdk2, importin-alpha and NuMA is required for H2O2-induced mitosis in primary type II pneumocytes. AB - Proliferation of alveolar type II pneumocytes, the multipotent stem cells of the alveoli, has been implicated in the development of lung adenocarcinoma. Hydrogen peroxide (H(2)O(2)), a potent promoter of signaling cascades, can mediate the transmission of many intracellular signals including those involved in cell proliferation. In this study using rat primary type II pneumocytes, we demonstrate that H(2)O(2) significantly increases mitosis through a pathway that includes cyclin-dependent kinase 2 (Cdk2); importin-alpha, a nuclear trafficking regulator; and nuclear mitotic apparatus protein (NuMA), an essential component in mitotic spindle pole formation. Upon H(2)O(2) treatment, Cdk2 is phosphorylated at position thr-160 leading to increases in importin-alpha and NuMA protein levels and resulting in a significant increase of G(2)/M phase in a roscovatine-dependent manner. Type II pneumocytes transfected with NuMA cDNA also show significant increases in G(2)/M phase, NuMA, Cdk2 thr-160 and importin-alpha expression. These effects were prevented by catalase. These results demonstrate that H(2)O(2) orchestrates a complex signaling network regulating S phase entry, nuclear trafficking and spindle pole formation through activation of Cdk2, importin-alpha, and NuMA. This pathway is essential for H(2)O(2)-induced mitosis in type II pneumocytes. PMID- 12729927 TI - Lipid-protein interactions in biological membranes: a structural perspective. AB - Lipid molecules bound to membrane proteins are resolved in some high-resolution structures of membrane proteins. An analysis of these structures provides a framework within which to analyse the nature of lipid-protein interactions within membranes. Membrane proteins are surrounded by a shell or annulus of lipid molecules, equivalent to the solvent layer surrounding a water-soluble protein. The lipid bilayer extends right up to the membrane protein, with a uniform thickness around the protein. The surface of a membrane protein contains many shallow grooves and protrusions to which the fatty acyl chains of the surrounding lipids conform to provide tight packing into the membrane. An individual lipid molecule will remain in the annular shell around a protein for only a short period of time. Binding to the annular shell shows relatively little structural specificity. As well as the annular lipid, there is evidence for other lipid molecules bound between the transmembrane alpha-helices of the protein; these lipids are referred to as non-annular lipids. The average thickness of the hydrophobic domain of a membrane protein is about 29 A, with a few proteins having significantly smaller or greater thicknesses than the average. Hydrophobic mismatch between a membrane protein and the surrounding lipid bilayer generally leads to only small changes in membrane thickness. Possible adaptations in the protein to minimise mismatch include tilting of the helices and rotation of side chains at the ends of the helices. Packing of transmembrane alpha-helices is dependent on the chain length of the surrounding phospholipids. The function of membrane proteins is dependent on the thickness of the surrounding lipid bilayer, sometimes on the presence of specific, usually anionic, phospholipids, and sometimes on the phase of the phospholipid. PMID- 12729928 TI - Effects of alterations of the amino-terminal glycine of influenza hemagglutinin fusion peptide on its structure, organization and membrane interactions. AB - Mutations of the glycine residue at the amino terminus of HA2 have been shown to have a large effect on the fusion activity of HA2, the extent of which apparently correlates with the side chain bulkiness of the substituting amino acids. To investigate into the cause of abrogation in fusogenicity and virus-promoted fusion mechanism, we synthesized several peptides in which this glycine was substituted by serine, glutamic acid, or lysine. 1,2-Dimyristoyl-sn-glycero-3 phosphocholine (DMPC) and 1,2-dimyristoyl sn-glycero-3-phosphoglycerol (DMPG) were used as model membranes in the fluorescence, circular dichroism (CD), and FTIR measurements while sodium dodecyl sulfate was used in NMR studies. We found that, for the less active variants, affinity to membrane, degree of solvent dehydration, lipid perturbation, depth of insertion, and helicity were less. Comparison of affinity to membrane bilayer among these analogs revealed that binding of the fusion peptide is determined largely by the hydrophobic effect. Additionally, the orientation is closer to the membrane normal for the wild-type fusion peptide in the helix form while the inactive analogs inserted more parallel to the membrane surface. PMID- 12729929 TI - Investigating magnetically aligned phospholipid bilayers with various lanthanide ions for X-band spin-label EPR studies. AB - This paper reports the EPR spectroscopic characterization of a model membrane system that magnetically aligns with a variety of different lanthanide ions in the applied magnetic field (<1 T) of an X-band EPR spectrometer. The ability to align phospholipid bilayer systems is valuable because the anisotropic spectra provide a more detailed and complete description of the structural and motional properties of the membrane-associated spin label when compared to randomly dispersed EPR spectra. The nitroxide spin probe 3beta-doxyl-5alpha-cholestane (cholestane or CLS) was inserted into the bilayer discs to demonstrate the effects of macroscopic bilayer alignment through the measurement of orientational dependent hyperfine splittings. The effects of different lanthanide ions with varying degrees of magnetic susceptibility anisotropy and relaxation properties were examined. For X-band EPR studies, the minimal amounts of the Tm(3+), Yb(3+), and Dy(3+) lanthanide ions needed to align the phospholipid bilayers were determined. Power saturation EPR experiments indicate that for the sample compositions described here, the spin-lattice relaxation rate of the CLS spin label was increased by varying amounts in the presence of different lanthanide (Gd(3+), Dy(3+), Er(3+), Yb(3+), and Tm(3+)) ions, and in the presence of molecular oxygen. The addition of Gd(3+) caused a significant increase in the spin-lattice relaxation rate of CLS when compared to the other lanthanide ions tested. PMID- 12729930 TI - Characterization of the interactions between Nedd4-2, ENaC, and sgk-1 using surface plasmon resonance. AB - Previous studies have characterized interactions between the ubiquitin ligase Nedd4-1 and the epithelial Na(+) channel (ENaC). Such interactions control the channel cell surface expression and activity. Recently, evidence has been provided that a related protein, termed Nedd4-2, is likely to be the true physiological regulator of the channel. Unlike Nedd4-1, Nedd4-2 also interacts with the aldosterone-induced channel activating kinase sgk-1. The current study uses surface plasmon resonance to quantify the binding of the four WW domains of Nedd4-2 to synthetic peptides corresponding to the PY motifs of ENaC and sgk-1. The measurements demonstrate that WW3 and WW4 are the only Nedd4-2 domains interacting with both ENaC and sgk-1 and that their binding constants are in the 1-6 microM range. PMID- 12729931 TI - Single-step immunoaffinity purification and characterization of dodecylmaltoside solubilized human neutrophil flavocytochrome b. AB - Flavocytochrome b (Cyt b) is a heterodimeric, integral membrane protein that serves as the central component of an electron transferase system employed by phagocytes for elimination of bacterial and fungal pathogens. This report describes a rapid and efficient single-step purification of Cyt b from human neutrophil plasma membranes by solubilization in the nonionic detergent dodecylmaltoside (DDM) and immunoaffinity chromatography. A similar procedure for isolation of Cyt b directly from intact neutrophils by a combination of heparin and immunoaffinity chromatography is also presented. The stability of Cyt b was enhanced in DDM relative to previously employed solubilizing agents as determined by both monitoring the heme spectrum in crude membrane extracts and assaying resistance to proteolytic degradation following purification. Gel filtration chromatography and dynamic light scattering indicated that DDM maintains a predominantly monodisperse population of Cyt b following immunoaffinity purification. The high degree of purity obtained with this isolation procedure allowed for direct determination of a 2:1 heme to protein stoichiometry, confirming previous structural models. Analysis of the isolated heterodimer by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry allowed for accurate mass determination of p22(phox) as indicated by the gene sequence. Affinity-purified Cyt b was functionally reconstituted into artificial bilayers and demonstrated that catalytic activity of the protein was efficiently retained throughout the purification procedure. PMID- 12729932 TI - Hyaluronan is a key component in cryoprotection and formulation of targeted unilamellar liposomes. AB - Lyophilized unilamellar liposomes (ULV), the dosage form of choice for shelf life, revert upon reconstitution to the larger multilamellar liposomes (MLV), which is detrimental to the many carrier-mediated therapies that require small particles. High doses of sugars such as trehalose, sucrose and others, included in the original formulations for cryoprotection, were shown to prevent the conversion to MLV. In this study we set out to test whether hyaluronan (HA), the surface-bound ligand in our previously developed targeted bioadhesive liposomes (BAL), can also act as a cryoprotectant. The studies included structural and physicochemical characterization of original and reconstituted hyaluronan-ULV (HA ULV). For each HA-ULV, similar regular ULV (RL-ULV) served as controls. Four properties were tested: particle size, zeta potential, encapsulation efficiency and half-life of drug release (tau(1/2)), for three drugs-chloramphenicol (CAM), vinblastine (VIN) and mitomycin C (MMC). Encapsulation efficiencies of the original systems were quite alike for similar RL-ULV and HA-ULV ranging from 25% to 70%. All systems acted as sustained-release drug depots, tau(1/2) ranging from 1.3 to 5.3 days. Drug species and lipid composition were the major determinants of encapsulation and release magnitudes. By all tests, as anticipated, lyophilization generated significant changes in the reconstituted RL-ULV: 17-fold increase in diameter; tripling of zeta potential; 25-60% drop in encapsulation efficiencies; 25-30% decrease in tau(1/2). In contrast, the reconstituted HA-ULV retained the same dimensions, zeta potentials, encapsulation efficiencies and tau(1/2) of the original systems. These data clearly show HA to be a cryoprotectant, adding another clinically relevant advantage to HA-BAL. We propose that, like the sugars, HA cryoprotects by providing substitute structure stabilizing H-bonds. PMID- 12729933 TI - Effects of sphingomyelin on melittin pore formation. AB - The effect of sphingomyelin (SM), one of the main lipids in the external monolayer of erythrocyte plasma membrane, on the ability of the hemolytic peptide melittin to permeabilize liposomes was investigated. The peptide induced contents efflux in large unilamellar vesicles (LUV) composed of 1-palmitoyl-2 oleoylphosphatidylcholine (POPC)/SM (1:1 mole ratio), at lower (>1:10,000) peptide-to-lipid mole ratios than in pure POPC (>1:1000) or POPC/1-palmitoyl-2 oleoylphosphatidylglycerol (POPG) (1:1 mole ratio) (>1:300) vesicles. Analysis of the leakage data according to a kinetic model of pore formation showed a good fit for hexameric-octameric pores in SM-containing vesicles, whereas mediocre fits and lower surface aggregation constants were obtained in POPC and POPC/POPG vesicles. Disturbance of lateral separation into solid (s(o)) and liquid disordered (l(d)) phases in POPC/SM mixtures increased the peptide-dose requirements for leakage. Inclusion of cholesterol (Chol) in POPC/SM mixtures under conditions inducing lateral separation of lipids into liquid-ordered (l(o)) and l(d) phases did not alter the number of melittin peptides required to permeabilize a single vesicle, but increased surface aggregation reversibility. Partitioning into liposomes or insertion into lipid monolayers was not affected by the presence of SM, suggesting that: (i) melittin accumulated at comparable doses in membranes with different SM content, and (ii) differences in leakage were due to promotion of melittin transmembrane pores under coexistence of s(o) l(d) and l(o)-l(d) phases. Our results support the notion that SM may regulate the stability of size-defined melittin pores in natural membranes. PMID- 12729934 TI - Escherichia coli lacking the AcrAB multidrug efflux pump also lacks nonproteinaceous, PHB-polyphosphate Ca2+ channels in the membrane. AB - PHB(polyP) complexes bind calcium and form calcium channels in the cytoplasmic membrane in Escherichia coli and are likely to be important in Ca(2+) homeostasis in this organism. E. coli N43, which lacks the AcrA component of a major multidrug resistance pump, was shown to be defective in calcium handling, with an inability to maintain submicromolar levels of free Ca(2+) in the cytoplasm. Therefore, using an N-phenyl-1-napthylamine (NPN)-dependent fluorescence assay, we measured temperature-dependent phase transitions in the membranes of intact cells. These transitions specifically depend on the presence of PHB(Ca(2+)polyP) complexes. PHB(Ca(2+)polyP) channel complexes, particularly in stationary phase cultures, were detected in wild-type strains; however, in contrast, isogenic acrA(-) strains had greatly reduced amounts of the complexes. This indicates that the AcrAB transporter may have a novel, hitherto undetected physiological role, either directly in the membrane assembly of the PHB complexes or the transport of a component of the membrane, which is essential for assembly of the complexes into the membrane. In other experiments, we showed that the particular defective calcium handling detected in N43 was not due to the absence of AcrA but to other unknown factors in this strain. PMID- 12729935 TI - Intracellular delivery of ceramide lipids via liposomes enhances apoptosis in vitro. AB - Ceramide lipids have emerged as important intracellular signalling molecules that mediate diverse cellular effects, of which programmed cell death, or apoptosis, has attracted significant interest. Although the exact mechanism(s) by which ceramides trigger apoptosis is not fully understood, there is considerable evidence that they are key mediators of this response. Exogenously applied, cell permeable ceramides have been shown to induce apoptosis when incubated with cells in culture. We examined here the cytotoxicity of ceramides with varying acyl chain lengths in order to determine whether acyl chain length affects pro apoptotic activity within the concentration range of 0-100 microM. We found that for C(6)-, C(8)-, C(10)-, C(14)- and C(16)-ceramide, the chain length was inversely proportional to cytotoxic activity, with C(6)-ceramide being most active (IC(50) values in the 3-14 microM range) and C(16)-ceramide being least active (IC(50) values in excess of 100 microM) in the MDA435/LCC6 human breast cancer and J774 mouse macrophage cell lines investigated. Using these two ceramide forms we were able to correlate the observed cytotoxicity with cellular uptake, and we observed that a lack of intracellular delivery may be responsible for the weak activity of C(16)-ceramide. We therefore investigated the possibility of incorporating ceramide lipids into liposome bilayers to enhance this delivery. We demonstrate that stable, ceramide-containing liposomes can be formulated, and that they are cytotoxic when taken up by cells in vitro. These results provide an increased understanding of the differences in cytotoxic activity of exogenous short- and long-chain ceramide lipids, and their incorporation into biologically active liposomal formulations opens new avenues for apoptosis induction. PMID- 12729936 TI - Modulation of Ca2+-activated K+ channels of human erythrocytes by endogenous protein kinase C. AB - Single IK(Ca) channels of human erythrocytes were studied with the patch-clamp technique to define their modulation by endogenous protein kinase C (PKC). The perfusion of the cytoplasmic side of freshly excised patches with the PKC activator, phorbol 12-myristate 13-acetate (PMA), inhibited channel activity. This effect was blocked by PKC(19-31), a peptide inhibitor specific for PKC. Similar results were obtained by perfusing the membrane patches with the structurally unrelated PKC activator 1-oleoyl-2-acetylglycerol (OAG). Blocking of this effect was induced by perfusion with PKC(19-31) or chelerythrine. Channel activity was not inhibited by the PMA analog 4alpha-phorbol 12,13-didecanoate (4alphaPDD), which has no effect on PKC. Activation of endogenous cAMP-dependent protein kinase (PKA), which is known to up-modulate IK(Ca) channels, restored channel activity previously inhibited by OAG. The application of OAG induced a reversible reduction of channel activity previously up-modulated by the activation of PKA, indicating that the effects of the two kinases are commutative, and antagonistic. Kinetic analysis showed that down-regulation by PKC mainly changes the opening frequency without significantly affecting mean channel open time and conductance. These results provide evidence that an endogenous PKC down-modulates the activity of native IK(Ca) channels of human erythrocytes. Our results show that PKA and PKC signal transduction pathways integrate their effects, determining the open probability of the IK(Ca) channels. PMID- 12729937 TI - N-terminal insertion of alamethicin in channel formation studied using its covalent dimer N-terminally linked by disulfide bond. AB - Alamethicin is supposed to form helix-bundle-type channels by inserting the N terminus into bilayer lipid membranes under sufficient voltages. The N-terminal insertion has been studied with an alamethicin dimer (di-alm) N-terminally linked by a disulfide bond and by the asymmetric addition of dithiothreitol (DTT) and tetrathionate (TT) to the membrane. When di-alm was added to the cis-side membrane, it forms long-lasting channels with the lifetime tau of about 100 ms at cis-positive voltages. The lifetime was reduced to a few milliseconds by addition of DTT to the cis-side membrane, indicating that most of the channels were formed by the monomers (alm-SH) that resulted from the cleavage of the disulfide bond in di-alm. The succeeding addition of TT to the trans-side produced channels of tau=10-20 ms besides the channels of alm-SH. The results suggested that TT reacted with the N-terminal thiol group of alm-SH located at the trans-side of the membrane to alter the lifetime. The N-terminal insertion of alamethicin helices by voltage activation, therefore, was confirmed. PMID- 12729938 TI - Effect of controlled-release melatonin on sleep quality, mood, and quality of life in subjects with seasonal or weather-associated changes in mood and behaviour. AB - This study aimed to explore the effects of melatonin on sleep, waking up and well being in subjects with varying degrees of seasonal or weather-associated changes in mood and behaviour. Fifty-eight healthy adults exhibiting subsyndromal seasonal affective disorder (s-SAD) and/or the negative or positive type of weather-associated syndrome (WAS) were randomised to either 2 mg of sustained release melatonin or placebo tablets 1-2 h before a desired bedtime for 3 weeks. Outcome measures were changes from baseline in sleep quality, sleepiness after waking, atypical depressive symptoms and health-related quality of life by week three. Early morning salivary melatonin concentrations were measured at baseline and treatment cessation in all subjects. Melatonin administration significantly improved the quality of sleep (P=0.03) and vitality (P=0.02) in the subjects with s-SAD, but attenuated the improvement of atypical symptoms and physical parameters of quality of life compared to placebo in the subjects with WAS, positive type. PMID- 12729939 TI - Interaction between angiotensin-converting enzyme and catechol-O methyltransferase genotypes in schizophrenics with poor response to conventional neuroleptics. AB - Angiotensin-converting enzyme (ACE) modulates dopamine turnover in the brain and catechol-O-methyltransferase (COMT) enzyme is an important agent in the metabolic inactivation of dopamine and norepinephrine. Functional polymorphism in the COMT and ACE genes causes variation in enzyme activities. We investigated the relationship of COMT and ACE gene polymorphism with response to conventional neuroleptic treatment in schizophrenia. In this study population we had earlier detected that COMT genotype is associated with unsatisfactory drug response. A total of 94 schizophrenic patients were evaluated either as responders (n=43) or non-responders (n=51). The responders had experienced a fair and steady response to conventional neuroleptics. The non-responders had failed to achieve an acceptable response to conventional neuroleptics. We also used a control population of 94 age- and gender-matched blood donors. Genotyping of the COMT and ACE genes was performed by polymerase chain reaction. The risk of having both low activity COMT and high activity ACE genotypes was over 10 times higher (odds ratio=10.89, 95%CI 1.14-103.98, P=0.04) in the non-responders compared to responders. ACE genotype alone did not differ between any groups. This finding may suggest a possible interaction with low activity COMT and high activity ACE genotype in association with poor response to conventional neuroleptics. PMID- 12729940 TI - 5-HT1A responsivity in patients with panic disorder before and after treatment with aerobic exercise, clomipramine or placebo. AB - Blunted neuroendocrine and physiological responses to the selective 5-HT(1A) receptor agonist, ipsapirone, have been observed in patients with panic disorder and/or agoraphobia (PDA). In order to examine whether this hyporesponsiveness to ipsapirone is modified by pharmacological or non-pharmacological therapeutic interventions, challenges with an oral dose of ipsapirone (0.3 mg/kg) and placebo were performed in patients with PDA before and after 10 weeks of treatment with clomipramine, aerobic exercise and placebo. Before treatment, administration of ipsapirone was followed by significant increases of cortisol, anxiety and other psychopathological symptoms in comparison to the placebo challenge. In addition, a significant decrease of body temperature was observed. After the 10-week treatment period, the psychological responses to ipsapirone were significantly reduced in the clomipramine and the exercise group. In contrast, there was a non significant trend towards higher cortisol responses after clomipramine and exercise treatment. The hypothermic response to ipsapirone was significantly reduced by clomipramine treatment. In conclusion, our results demonstrate that effective treatment of panic disorder has divergent effects on the psychological, neuroendocrine and temperature responses to ipsapirone. PMID- 12729942 TI - Chronic lithium treatment with or without haloperidol fails to affect the morphology of the rat cerebellum. AB - We used unbiased stereological principles to determine whether long-term administration of lithium at human therapeutic levels, with or without haloperidol, affects the number or sizes of cerebellar Purkinje cells or the volume of histological layers in the rat cerebellum. Twenty-eight rats were randomly divided into three groups, receiving either no treatment, lithium, or lithium combined with haloperidol. The serum lithium levels ranged from 0.50 to 0.77 mmol/l. Haloperidol was given at a daily dose of 1 mg/kg. After 30 weeks of treatment, the animals were killed and the cerebelli were histologically prepared. No statistically significant differences were observed between the groups with respect to the cerebellar measures. PMID- 12729941 TI - Adenosine receptor agonists or antagonists alter antinociception, but did not show an interaction with imipramine-induced antinociception in the formalin test in mice. AB - In this study, the antinociceptive effect of imipramine and adenosine agents, and interactions between imipramine with adenosine drugs in mice in the formalin test, have been investigated. Intraperitoneal administration of different doses of imipramine (10, 20, 30 and 40 mg/kg) induced a dose dependent antinociception in mice, in both the first and second phases of the formalin test. The adenosine A(1) receptor agonists, R-(N(6)-phenylisopropyl)-adenosine (0.015, 0.03 and 0.1 mg/kg) and 5'-N-ethylcarboxamide adenosine (0.001, 0.005, 0.01 mg/kg), but not 2 chloroadenosine (0.1 and 0.5 mg/kg), and the adenosine receptor antagonist, 8 phenyltheophylline (0.1, 0.5 and 1 mg/kg), but not 1,3-dipropyl-7-methyl-xanthine (0.5 and 5 mg/kg), also produced an antinociceptive response. Lower dose of the adenosine receptor antagonist theophylline induced antinociception, while a higher dose of the drug caused hyperalgesia. Theophylline reduced the response induced by imipramine. It is concluded that adenosine systems are not involved in imipramine responses in the formalin test. PMID- 12729943 TI - Anti-cataleptic effects of clozapine, but not olanzapine and quetiapine, on SCH 23390- or raclopride-induced catalepsy in rats. AB - The present study investigated potential anti-cataleptic properties of the prototype atypical antipsychotic clozapine and two newly developed atypical antipsychotics, olanzapine and quetiapine, which are structurally related and display similar pharmacological profiles to clozapine. Clozapine (2.5 mg kg(-1), s.c.), but not olanzapine (2.0 mg kg(-1), s.c.) and quetiapine (20.0 mg kg(-1), s.c.), blocked catalepsy induced either by the dopamine D(1/5) receptor antagonist SCH 23390 (50.0 microg kg(-1), s.c) or the selective dopamine D(2/3) receptor antagonist raclopride (4.0 mg kg(-1), s.c.). Such findings are consistent with the beneficial effects of clozapine in the management of drug induced psychosis in parkinsonian patients, and suggest that neither olanzapine nor quetiapine may be a safe alternative to clozapine in this field. Furthermore, the results indicate that clozapine has a unique pharmacological profile that distinguishes it from olanzapine and quetiapine. The mechanisms underlying anti cataleptic or anti-parkinsonian properties of clozapine are unclear but may be related to dopamine D(1) receptor agonism of clozapine. PMID- 12729944 TI - DRD4 exon III polymorphism and response to risperidone in Israeli adolescents with schizophrenia: a pilot pharmacogenetic study. AB - This study examined the possible association between the polymorphism in the dopamine receptor DRD4 gene and response to risperidone among 24 Israeli Jewish adolescent inpatients with first-episode schizophrenia. Response was categorically determined by a change of >40% on the Brief Psychiatric Rating Scale (BPRS). No significant association was found between the DRD4 genotype and clinical response, although carriers of <7 repeat alleles demonstrated higher response rate (10/20 vs. 0/4, P=0.11). Studies in larger groups of adolescent schizophrenia patients are warranted to clarify the possible association between DRD4 exon III repeat alleles and the response to risperidone. PMID- 12729945 TI - Neonatal lesions in the amygdala or ventral hippocampus disrupt prepulse inhibition of the acoustic startle response; implications for an animal model of neurodevelopmental disorders like schizophrenia. AB - Prepulse inhibition of the acoustic startle response is a behavioural tool applied to assess sensorimotor gating processes in humans and rats. Schizophrenic patients show deficits in prepulse inhibition of the acoustic startle response. The animal model of neurodevelopmental disorders such as schizophrenia, as purported in earlier reports and the present study, is based on the assumption that damage to brain structures early in life (on day 7) disrupts brain maturation of structures connected to the damaged areas, measurable by behavioural changes, whereas similar damage later in life (on day 21) does not result in these behavioural changes. Locomotor activity, the acoustic startle response and its prepulse inhibition were investigated in adult rats lesioned in the amygdala or ventral hippocampus on day 7 or 21 of life. The acoustic startle response was increased in animals lesioned in the amygdala on day 7 or 21 of life, but not in animals lesioned in the ventral hippocampus. Prepulse inhibition was impaired and locomotor activity enhanced in animals lesioned in the amygdala or ventral hippocampus on day 7, but not in animals lesioned in these structures on day 21 of life. The results on the acoustic startle response are suggestive of amygdaloid influences on modulation of the acoustic startle response. The effects of early postnatal lesions on prepulse inhibition and locomotor activity are in support of the animal model of neurodevelopmental disorders like schizophrenia. PMID- 12729946 TI - Chronic lithium and sodium valproate both decrease the concentration of myoinositol and increase the concentration of inositol monophosphates in rat brain. AB - One of the mechanisms underlying lithium's efficacy as a mood stabilizer in bipolar disorder has been proposed to be via its effects on the phosphoinositol cycle (PI cycle), where it is an inhibitor of the enzyme converting inositol monophosphates to myoinositol. In contrast, sodium valproate, another commonly used mood stabilizer, appears to have no direct effects on this enzyme and was thus believed to have a different mechanism of action. In the present study, high resolution nuclear magnetic resonance (NMR) spectroscopy was used to study the chronic effects of both lithium and sodium valproate on the concentrations of myoinositol and inositol monophosphates in rat brain. As predicted, lithium treated rats exhibited a significant increase in the concentration of inositol monophosphates and a significant decrease in myoinositol concentration compared to saline-treated controls. However, unexpectedly, sodium valproate administration produced exactly the same results as lithium administration. These novel findings suggest that both lithium and sodium valproate may share a common mechanism of action in the treatment of bipolar disorder via actions on the PI cycle. PMID- 12729947 TI - Effects of 8-OH-DPAT on open field performance of young and aged rats prenatally exposed to diazepam: a tool to reveal 5-HT1A receptor function. AB - Central GABAergic and serotoninergic systems interact with one another and are implicated in controlling different behaviours. A gentle early long-lasting handling can prevent the deficits in locomotion and exploration in open field (O.F.) in 3-month-old male rats prenatally exposed to diazepam (DZ). Purpose of this study was to extend the research to older handled rats prenatally exposed to DZ and to assess the activity of 5-HT1A receptors (Rs), evaluating the performance in O.F. at 3 and 18 months of age following 8-OH-DPAT administration. A single daily s.c. injection of DZ (1.5 mg/kg) from gestation day 14 to gestation day 20 induced in aged, but not in young rats, a decrease in total distance travelled (TDT) and in rearing frequency (RF) and an increase of transitions from the periphery to the centre of the arena (CNT) and in the time spent in the centre of the arena (CAT), compared to controls. 8-OH-DPAT (0.150 mg/kg s.c.), given 1 h before testing, increased TDT and decreased RF, CNT and CAT in both vehicle- and DZ-exposed young rats. In aged rats prenatally exposed to DZ, 8-OH-DPAT induced an increase in TDT and a slight decrease in RF, CNT and CAT. These findings indicate that the effects of handling and of 8-OH-DPAT in prenatally DZ-exposed rats are age-dependent and suggest that O.F. test can represent a valid tool to identify the changes in 5-HT1A Rs activity following drug treatment. PMID- 12729949 TI - Spike sequences and mean firing rate in rat neocortical neurons in vitro. AB - Properties of neuronal discharges and their interrelationship with mean evoked firing rate (MF) were investigated in detail in the rat somatosensory cortex in vitro. Firing was evoked by 1-min depolarizing current injection (0.1-1.0 nA) into pyramidal neurons (n=93) of layers II/III and V. Spike sequence patterns (SSPs) with consecutive increasing or decreasing interspike intervals were analyzed. SSP rate (change of pattern, 2-50 min(-1)) and the deviations of afterhyperpolarizing potentials (up to 1.5 mV) associated with the SSPs were significantly correlated with MF. The number of transitions in spike frequency was dependent on the MF and showed a biphasic interrelation with a reversible point at 5-8 Hz. All neurons analysed were subdivided into two groups: with depolarizing potential after single spike and without such afterdepolarizing potential. It was found that despite significant differences in electrophysiological properties of the cells from these groups, similar dependency in patterns of their discharges upon MF was observed. A possible role of spike sequence patterns in neocortical rhythmical processes is discussed. PMID- 12729948 TI - Alterations in cellular IRP-dependent iron regulation by in vitro manganese exposure in undifferentiated PC12 cells. AB - Manganese (Mn) may interfere with iron regulation by altering the binding of iron regulatory proteins (IRPs) to their response elements found on the mRNA encoding proteins critical to iron homeostasis. To explore this, the effects of 24-h in vitro manganese exposure (1, 10, 50, and 200 microM Mn) on: (i) total intracellular and labile iron concentrations; (ii) the cellular abundance of transferrin receptor (TfR), H- and L-ferritin, and mitochondrial aconitase proteins; and (iii) IRP binding to a [32P](-) labeled mRNA sequence of L-ferritin were evaluated in undifferentiated PC12 cells. In vitro manganese exposure altered the cellular abundance of TfR, H-/L-ferritin, and m-aconitase, resulting in an increase in labile iron. This latter effect led to a decrease in IRP binding activity at the lower (10 and 50 microM) manganese exposures. In contrast, 200 microM manganese exposure increased IRP binding, in spite of the significant increase in labile iron. These data indicate that at lower exposures, manganese directly interfered with IRP-dependent translational events, producing an increase in labile iron, which in turn signaled a decrease in IRP binding at 24 h. At higher exposures, the intracellular burden of manganese resulted in overt cytotoxicity and appeared to compromise the normal compensatory response to increased labile iron, producing increased IRP binding. We conclude that low to moderate manganese exposure interferes with cellular iron regulation, and thus may serve as a contributory mechanism underlying manganese neurotoxicity. PMID- 12729950 TI - Tolerance of neonatal rat brain to acute hyperammonemia. AB - The aim of the present work was to study the effects of hyperammonemia on brain energy metabolism in neonatal rats. Rats were rendered hyperammonemic by ammonium acetate administration. This decreased brain ATP concentrations but enhanced brain ammonia and lactate levels in both adult and neonatal rats. In adult rats, the decrease in brain ATP concentrations was accompanied by a plunge in the respiratory control rate (RCR) of brain mitochondria. However, the ammonia induced effect on RCR was not observed in neonatal rats, suggesting that the fall in ATP levels observed in neonatal rats would not be due to an impairment of mitochondrial respiratory efficiency. However, in neonatal rats the increase in blood and brain ammonia concentrations did not change brain glutamate concentrations but decreased glutamine contents. These results may be of relevance for the understanding of the resistance of neonatal rats observed in this work to acute ammonia toxicity PMID- 12729951 TI - Spinal cord stimulation inhibits long-term potentiation of spinal wide dynamic range neurons. AB - It has been suggested that long-term potentiation (LTP) of dorsal horn neurons is a phenomenon that contributes to the development of chronic neuropathic pain. Spinal cord stimulation (SCS) may be an effective tool in alleviating such pain. The aim of this electrophysiological study in rats was to examine if SCS suppresses LTP of dorsal horn wide dynamic range (WDR) neurons. Increased knowledge of the mechanisms behind the effects of SCS may facilitate its further advancement and improve clinical efficacy. As previously shown, intensive, high frequency electrical stimulation of the sciatic nerve in the rat induces an increased firing response of WDR neurons. Here we report that SCS gradually reduced this increased C-fiber response back to the baseline level. However, A fiber responses were neither potentiated by the conditioning stimulus used nor were they affected by SCS. These data suggest that SCS affects the C-fiber component of dorsal horn central sensitization which is noteworthy since SCS, based on previous studies, is believed to primarily influence A-fiber functions. PMID- 12729952 TI - Active decay of composite excitatory postsynaptic potentials in hippocampal slices from young rats. AB - NMDA receptor dependent synaptic plasticity was examined in hippocampal slices using a novel pharmacological pairing procedure. Field excitatory postsynaptic potentials (EPSPs) were recorded from the CA1 area of slices maintained in a low Mg(2+) solution using a stimulus rate of 0.1-0.2 Hz. The NMDA receptor antagonist 2-amino-5-phosphonovalerate (AP5) was initially included in the perfusion solution to establish baseline recording of isolated AMPA EPSPs. Washing out AP5 led to the expression of composite EPSPs, containing both AMPA and NMDA receptor mediated components. Following an initial, transient potentiation of the AMPA component, the composite responses gradually decayed for several hours, involving AMPA and NMDA components to a similar extent. This decay was input specific and could be terminated at any stage by reapplication of AP5. Subsequent long-term potentiation (LTP) reversed the effect to an extent inversely related to the degree of depression. Experiments to test the interaction with long-term depression (LTD) revealed a significant but incomplete overlap between the two depression processes. In conclusion, pairing synaptic activation at test stimulus frequency with pharmacological unblocking of NMDA receptors allows for expression of composite EPSPs that decay substantially, due to an active mechanism. The underlying process appears to be at least partly distinct from those involved in homosynaptic LTP and LTD. PMID- 12729953 TI - Melatonin synthesis in retina: circadian regulation of arylalkylamine N acetyltransferase activity in cultured photoreceptor cells of embryonic chicken retina. AB - The key regulatory enzyme in melatonin synthesis is arylalkylamine N acetyltransferase (AANAT). In vivo, AANAT activity in chicken retinal photoreceptor cells exhibits a circadian rhythm that peaks at night. The purpose of the present study was to investigate the temporal development of light/dark and circadian oscillations of AANAT activity in cultured retinal cells prepared from 6- and 8-day-old chicken embryos (E6, E8, respectively). Photoreceptor cells prepared from E6 retinas and incubated under a 14-h light/10-h dark (LD) cycle of illumination for 5-7 days displayed prominent daily fluctuations in AANAT activity on days 5 and 6 in vitro. However, when E6 cells, incubated for 5 days under LD, were transferred to continuous (24 h/day) darkness (DD) on day 6, no daily pattern of activity was observed. This result indicates that AANAT fluctuations were light-driven and not circadian at this stage. In contrast, cells prepared from E8 embryos and incubated under conditions identical to those for E6 cells displayed prominent rhythms of AANAT activity in both LD and DD, indicative of circadian control. To determine if circadian control of AANAT activity would develop in E6 cells incubated for a longer period of time to allow maturation, cells were incubated for 8 days in LD followed by 2 days in DD. AANAT activity in these cells was rhythmic in both LD and DD. In cells incubated in this manner, a 2-h light pulse in the middle of the subjective night suppressed AANAT activity, indicating that the enzyme activity in the cultured cells is acutely suppressed by light, as it is in vivo. These results indicate that the ability to express circadian regulation of AANAT activity is an intrinsic property of retinal cells that can develop in vitro. Development of light-dark regulation of AANAT activity appears to precede the circadian clock-control of enzyme activity. PMID- 12729954 TI - Quantification of prolactin-releasing peptide (PrRP) mRNA expression in specific brain regions of the rat during the oestrous cycle and in lactation. AB - Real-time Taqman RT-PCR was used to make quantitative comparisons of the levels of PrRP mRNA expression in micropunch brain samples from rats at different stages of the oestrous cycle and in lactation. The nucleus of the solitary tract and ventrolateral reticular nuclei of the medulla oblongata contained significantly (P<0.05) greater levels of PrRP mRNA than any hypothalamic region. Within the hypothalamus, the highest level of PrRP expression was localised to the dorsomedial aspect of the ventromedial hypothalamus. All other hypothalamic regions exhibited significantly (P<0.05) lower levels of expression, including the rostral and caudal dorsomedial hypothalamus. Very low levels of PrRP expression were observed in the arcuate nucleus, paraventricular nucleus, medial preoptic nucleus and ventrolateral aspect of the ventromedial hypothalamus. No significant changes in PrRP expression were noted in any sampled region between proestrus, oestrus or dioestrus. Similarly, PrRP expression in hypothalamic regions did not differ between lactating and non-lactating (dioestrous) animals. During validation of RT-PCR techniques we cloned and sequenced a novel splice variant of PrRP from the hypothalamus. This variant arises from alternative splicing of the donor site within exon 2, resulting in an insert of 64 base pairs and shift in the codon reading frame with the introduction of an early stop codon. In the hypothalamus and brainstem, mRNA expression of the variant was restricted to regions that expressed PrRP. These results suggest that PrRP expression in the hypothalamus may be more widespread than previously reported. However, the relatively low level of PrRP in the hypothalamus and the lack of significant changes in expression during the oestrous cycle and lactation provides further evidence that PrRP is unlikely to be involved in the regulation of prolactin secretion. PMID- 12729955 TI - The relationship between head size and intracranial volume in elderly subjects. AB - OBJECTIVE: To study the relationship between parenchymal head volume (PHV) and intracranial volume (ICV), and to compare the ability of these two measurements to reflect the association between maximum mature brain volume and late-life cognition. METHODS: An elderly sample of humans with a range of cognitive functions from normality, via mild cognitive impairment (MCI) to dementia (mean age 78.6, S.D. 2.8; mean MMSE 25.4, S.D. 4.2) was examined. Head-to-head measurements of ICV and parenchymal head volume (PHV) were obtained from three dimensional T1 weighted magnetic resonance images using automated procedures. Analyses of cognitive functions were based on continuous and categorial variables. RESULTS: PHV explained 55% of the variance in ICV. The ratio between PHV and ICV remained constant with increasing age and cognitive impairment. Measurements of PHV and ICV yielded comparable correlations with global cognitive performance. Group differences over gender and cognitive states were equally present in ICV and PHV. The relative risks of cognitive impairment that were associated with either small ICV or PHV were comparable. CONCLUSIONS: Measures of PHV can be considered as useful estimates of ICV and cerebral volume reserve. PMID- 12729956 TI - Somatodendritic dopamine release in rat substantia nigra influences motor performance on the accelerating rod. AB - The physiological role of somatodendritic dopamine release in the rat substantia nigra was evaluated with a combination of dual probe microdialysis and simultaneous motor performance tests on an accelerating rod. Three main findings support a modulating influence of somatodendritic dopamine release on motor coordination. (1) The rod performance tests were associated with an increase in extracellular dopamine but not 5-hydroxytryptamine concentrations in substantia nigra and with increases in both dopamine and 5-hydroxytryptamine concentrations in the striatum. (2) Nigral application of dopamine antagonists without intrinsic activity resulted in changed performances on the accelerating rod. The response to nigral perfusion with low concentrations (0.1, 1.0 microM) of the D(2)/D(3) antagonist raclopride consisted of an impairment in rod performance to 63% of the pre-perfusion performance. Higher concentrations (10, 100 microM), however, were not associated with impaired rod performance, but with increased striatal dopamine concentrations. Perfusion of the substantia nigra with 1, 10 and 100 microM of the D(1)/D(5)-antagonist SCH 23390 dose-dependently impaired rod performance. SCH 23390 consistently increased dopamine and 5-hydroxytryptamine concentrations in substantia nigra but did not change the dialysate in the striatum. (3) In unilaterally 6-hydroxydopamine-lesioned rats, a dose-dependent improvement in rod performance was observed during perfusion of the substantia nigra with the non-selective dopamine agonist apomorphine. PMID- 12729957 TI - Involvement of 5-hydroxytryptamine4 receptor in the exacerbation of neuronal loss by psychological stress in the hippocampus of SHRSP with a transient ischemia. AB - A transient forebrain ischemia produced a delayed neuronal death of the hippocampus pyramidal cells in stroke-prone spontaneously hypertensive rats (SHRSP). Long term exposure of rats to stress has been reported to induce deleterious effects on the brain including morphological neuronal degeneration in the hippocampus. The present study was designed to examine the effects of psychological and physical stress on the ischemia-related neuronal death and the effects of 5-hydroxytryptamine(4) (5-HT(4)) receptor antagonist. SHRSP were exposed to the psychological or physical stress for 60 min in the communication box once or repeatedly for 3 days and occluded. SB204070, a 5-HT(4) receptor antagonist was injected before the occlusion. Seven days after the occlusion, the number of the neurons damaged morphologically was examined. A transient bilateral carotid occlusion produced a neuronal death of the CA1 subfield of the hippocampus in a time-dependent manner between 3 and 10 min. A 4 min occlusion induced very little morphological damage and a 5 min one produced a significant neuronal death. Exposure of rats to the psychological stress during 60 min for 3 days before the ischemic insults damaged the pyramidal cells by 4 min ischemia much more than without stress. Physical stress daily for 3 times also increased the damaged neurons. Pretreatment of SB204070 0.1 mg/kg after the stress exposure for 3 days significantly decreased the neuronal damage exacerbated by the stress exposure; however, it did not alter the damage induced by 4 or 10 min occlusion without stress. These results suggest that the repeated exposure of animals to the stress dramatically exacerbates the neuronal death by a transient ischemia and the 5-HT(4) receptor may be involved in the stress-induced exacerbating mechanism of the neuronal damage. PMID- 12729958 TI - Conditioned stimulus-induced reinstatement of extinguished cocaine seeking in C57BL/6 mice: a mouse model of drug relapse. AB - The scarcity of mouse models for relapse to cocaine seeking has curtailed the study of genetic factors that may contribute to susceptibility for drug relapse. To contribute to the development of a new mouse model of drug relapse, C57BL/6 (B6) mice were trained to press a lever for infusions of cocaine (0.35 mg/kg, i.v.) on a fixed ratio 1 schedule of reinforcement during daily 2-h sessions. A light+tone stimulus complex was presented simultaneously with each cocaine infusion. Mice then underwent a series of extinction sessions during which lever presses had no scheduled consequences. As a result, lever pressing gradually declined. In experiment 1, the ability of the cocaine-paired light+tone stimulus complex to reinstate extinguished cocaine-seeking behavior (i.e. non-reinforced responses) was assessed. In experiment 2, the ability of cocaine priming (0, 1, 2.5, 5, 10, 20, and 40 mg/kg, i.p.) to reinstate cocaine-seeking behavior was measured. B6 mice failed to reinstate in response to i.p. cocaine priming; however, they exhibited robust conditioned stimulus-induced reinstatement. These findings suggest that conditioned stimulus-induced reinstatement in B6 mice is a promising model to study genetic and neurobiological factors that alter the ability of cocaine-paired stimuli to elicit relapse to cocaine seeking behavior. PMID- 12729959 TI - An investigation into the role of calcium in the modulation of rat synaptosomal D [3H]aspartate transport by docosahexaenoic acid. AB - The effect of the polyunsaturated fatty acid cis-4,7,10,13,16,19-docosahexaenoic acid (DHA) on the high-affinity, sodium-dependent uptake of D-[3H]aspartate into purified rat brain synaptosomes was examined. Incubation of the synaptosomes with 20 microM DHA caused over 50% inhibition of the maximum velocity (V(max)) of D [3H]aspartate transport. This inhibition was significantly potentiated by pre exposure of the synaptosomes to the fatty acid for 10 min prior to the start of the transport assay. Less highly unsaturated fatty acids such as arachidonic acid (cis-5,8,11,14-eicosatetraenoic acid), linolenic acid (cis-9,12,15 octadecatrienoic acid) and oleic acid (cis-9-octadecenoic acid) were significantly less potent than DHA. Removal of extracellular calcium, or reduction of the intracellular calcium concentration using the intracellular calcium chelator BAPTA/AM (10 microM), did not reduce the inhibition caused by DHA. On the other hand, an increase in the concentration of intracellular calcium mediated by thapsigargin (25 microM) or the calcium ionophore A23187 (10 or 100 nM) led to a reduction in the rate of D-[3H]aspartate transport in the absence of DHA. The CaM kinase II inhibitor, KN-93, reduced D-[3H]aspartate uptake independently of whether DHA was also present, but had no effect on the inhibition of D-[3H]aspartate uptake by either A23187 or thapsigargin. We conclude that whereas DHA inhibits synaptosomal D-[3H]aspartate uptake in a calcium-independent manner, a calcium-based mechanism exists that can also modulate glutamate transporter activity. PMID- 12729960 TI - Daily and circadian rhythms of neurotransmitters and related compounds in the hypothalamic suprachiasmatic nuclei of a diurnal vertebrate. AB - By using immunocytochemistry we tested whether neurotransmitters, and enzymes specific to neurotransmitters synthesis are rhythmically expressed in the suprachiasmatic nuclei of the hypothalamus of Ruin lizards Podarcis sicula either kept in light-dark cycles or constant darkness. Within the suprachiasmatic nuclei, prominent 24 h rhythms under 12:12 light-dark cycles were found for vasoactive intestinal polypeptide (VIP) and for tyrosine hydroxylase (TH). Peaks of both VIP and TH fell in the light phase of the cycle. Rhythmic expression of TH persisted under constant temperature and darkness, demonstrating the existence of circadian rhythms of TH in the suprachiasmatic nuclei. No rhythmic expression of neurotransmitters and related compounds was found in the periventricular nuclei, the supraoptic nuclei, and the rest of the hypothalamus. Our data are the first demonstration of rhythmic expression of neurotransmitters and related compounds in the suprachiasmatic nuclei of a non-mammalian vertebrate. The demonstration of a diurnal peak of VIP in a diurnal reptile-vs. nocturnal peak of VIP typical of nocturnal mammals-provides new information for comparative studies on the circadian physiology of the suprachiasmatic nuclei across vertebrate classes and their adaptation strategies to different temporal niches. PMID- 12729961 TI - Evidence for multiple mechanisms of kappa opioid tolerance in mesencephalic cultures. AB - Opioid tolerance limits the effectiveness of chronic opioid therapy. Kappa opioid tolerance in mesencephalic cultures has been established as a model system for evaluating neuronal mechanisms of tolerance. Our studies were carried out in primary cultures derived from the mesencephalon of rat embryos. [3H]Dopamine release assays were used to evaluate the kinetics of onset and recovery from tolerance to the kappa opioid receptor agonist (U69,593) which were measured with and without (1) modulators of cAMP-dependent kinase or (2) cycloheximide, an inhibitor of protein synthesis. Results were compared to the kinetics of recovery from kappa receptor alkylation by beta-chlornaltrexamine. The results showed that 8-bromo-cAMP and forskolin significantly accelerated the development of tolerance to U69,593. Rp-cAMP significantly slowed the onset of tolerance. Rates of recovery of cultures after 18 h of U69,593 were not significantly different regardless of whether cultures were incubated with U69,593 alone or in combination with 8-bromo-cAMP, forskolin or Rp-cAMP. Cycloheximide significantly slowed the recovery of cultures after 7 days of exposure to U69,593 but not after 18 h of exposure to U69,593. Recovery of cultures after alkylation of kappa receptors by beta-chlornaltrexamine was significantly slower than the recovery of cultures after 7 days of U69,593 exposure indicating that receptor synthesis is unlikely to be the major mechanism of recovery after prolonged opioid exposure. It is suggested that multiple mechanisms are responsible for tolerance to U69,593 in primary neuronal cultures. There are long lasting mechanisms of tolerance that do not involve destruction or permanent inactivation of cellular components. PMID- 12729962 TI - The effect of bicuculline application on azimuth-dependent recovery cycle of inferior collicular neurons of the big brown bat, Eptesicus fuscus. AB - The recovery cycle of auditory neurons is an important neuronal property, which determines a neuron's ability to respond to pairs of sounds presented at short inter-sound intervals. This property is particularly important for bats, which rely upon analysis of returning echoes to extract the information about targets after emission of intense orientation sounds. Because target direction often changes throughout the course of hunting, the changing echo direction may affect the recovery cycle and thus temporal processing of auditory neurons. In this study, we examined the effect of sound azimuth on the recovery cycle of inferior collicular (IC) neurons in the big brown bat, Eptesicus fuscus, under free-field stimulation conditions. Our study showed that the recovery cycle of most IC neurons (42/49, 86%) was longer when determined with sounds delivered at 40 degrees ipsilateral (i40 degrees ) than at 40 degrees contralateral (c40 degrees ) to the recording site. To study the contribution of GABAergic inhibition to sound azimuth-dependent recovery cycle, we compared the recovery cycle of IC neurons determined at two sound azimuths before and during iontophoretic application of bicuculline, an antagonist for GABA(A) receptors. Bicuculline application produced a greater decrease of the recovery cycle of these neurons at i40 degrees than at c40 degrees. As a result, the azimuth-dependent recovery cycle of these neurons was abolished or greatly reduced. Possible mechanisms underlying these observations and biological relevance to bat echolocation are discussed. PMID- 12729963 TI - Laminar expression of neurofilament protein in the superior colliculus of the marmoset monkey (Callithrix jacchus). AB - The expression profile of the monoclonal antibody SMI-32 was examined in the superior colliculus of adult marmosets. This antibody recognises subunits of the non- and dephosphorylated neurofilament protein, labelling predominantly neuronal perikarya and dendrites. The densest cellular label was observed in the intermediate layers (primarily, the stratum griseum intermediale), consisting of large multi- or bipolar neurones which were preferentially located within cytochrome oxidase-rich regions. The morphological characteristics of neurones showing heavy staining resemble those of extrinsic projection cells, suggesting a correlation between neurofilament content and axonal length. PMID- 12729965 TI - Marian Breland Bailey: many lives. (SQAB, May 25, 2002, Toronto, Canada). PMID- 12729964 TI - Retinal ganglion cells projecting to the dorsal raphe and lateral geniculate complex in Mongolian gerbils. AB - Injections of rhodamine-B into the dorsal raphe nucleus (DRN) and Fluoro-Gold into the lateral geniculate nucleus (LGN) revealed double-labeled retinal ganglion cells (DL RGCs) projecting to both nuclei. The soma-size distribution of DL RGCs was compared with three other distributions: DRN-projecting RGCs, LGN projecting RGCs, and a large sample of RGCs labeled via the optic nerve with DiI. DL RGC soma diameters fell primarily within the mid-to-upper size range of all three distributions. DL RGCs may provide information to both nuclei concerning comparable aspects of light and visual stimulation via collateralized axons. PMID- 12729966 TI - Stimulus representation in SOP: I. Theoretical rationalization and some implications. AB - THE SOP MODEL [INFORMATION PROCESSING IN ANIMALS: Memory Mechanisms, Erlbaum, Hillsdale, NJ, 1981, p. 5] is described in terms of its assumed stimulus representation, network characteristics, and rules for learning and performance. It is shown how several Pavlovian conditioning phenomena can be accounted on the basis of the model's presumed stimulus representation. Challenges to the SOP model prompted the adoption of a componential stimulus representation in: AESOP [Contemporary Learning Theories: Pavlovian Conditioning and the Status of Traditional Learning Theory, Erlbaum, Hillsdale, NJ, 1989, p. 149], this was a dual representation of the unconditioned stimulus (US), and C-SOP [Contemporary Learning: Theory and Application, Erlbaum, Mahwah, NJ, 2001, p. 23], this was a multi-component representation of the conditioned stimulus (CS). The assumption of a componential CS representation, where large numbers of elements can be separately learned about, necessitated a modification of the learning rule. The modified, "constrained" rule was found useful to explain timing characteristics of Pavlovian conditioned responses, as well as data offered by Rescorla [J. Exp. Psychol. Anim. Behav. Process. 26 (2000) 428; Q. J. Exp. Psychol. 54B (2001) 53; J. Exp. Psychol. Anim. Behav. Process. 28 (2002) 163] showing that stimuli trained in compound do not share the same quantitative fate. PMID- 12729967 TI - Stimulus representation in SOP: II. An application to inhibition of delay. AB - The componential extension of SOP accounts for conditioned response (CR) timing in Pavlovian conditioning by assuming that learning accrues with relative independence to stimulus elements that are differentially occasioned during the duration of the conditioned stimulus (CS). SOP, using a competitive learning rule and the assumption that temporal learning emerges via resolution of what is equivalent to an "AX+BX-" discrimination, predicts a progressive increase in the latency of the CR over training, or what Pavlov refer to as "inhibition of delay." Other componential models, which use noncompetitive learning rules, do not predict inhibition of delay. Either type of model makes the prediction indicated, independently of the length of the CS-unconditioned stimulus (US) interval. We report two experiments that demonstrated inhibition of delay when rabbits were trained with relatively long, but not with short, CS-US intervals. To account for this divergence, we assumed that the SOP stimulus trace involves two kinds of elements, some with a temporally distributed pattern of activity over the duration of the CS duration, and some with a randomly distributed pattern. This stimulus representation, not only allows for inhibition of delay with long but not short CS-US intervals, but in combination with SOP's performance rule deduces CR's with "Weber variability." PMID- 12729968 TI - MPR. AB - Mathematical Principles of Reinforcement (MPR) is a theory of reinforcement schedules. This paper reviews the origin of the principles constituting MPR: arousal, association and constraint. Incentives invigorate responses, in particular those preceding and predicting the incentive. The process that generates an associative bond between stimuli, responses and incentives is called coupling. The combination of arousal and coupling constitutes reinforcement. Models of coupling play a central role in the evolution of the theory. The time required to respond constrains the maximum response rates, and generates a hyperbolic relation between rate of responding and rate of reinforcement. Models of control by ratio schedules are developed to illustrate the interaction of the principles. Correlations among parameters are incorporated into the structure of the models, and assumptions that were made in the original theory are refined in light of current data. PMID- 12729969 TI - Mathematical principles of reinforcement and resistance to change. AB - Although Killeen's mathematical principles of reinforcement (MPR) apply to the asymptotic rate of a free operant after extended exposure to a single schedule of reinforcement, they can be extended to resistance to change in multiple schedules via alterations in the parameter representing the activating effects of reinforcers. MPR's predictions of resistance to change in relation to reinforcer rate on variable-interval (VI) schedules are empirically correct and agree with behavioral momentum theory (BMT). However, both MPR and BMT encounter problems in accounting for the effects of delayed reinforcement on resistance to change, relative to immediate reinforcement at the same rate. Further problems are raised by differences in resistance to change between variable-ratio (VR) and variable interval performances maintained by the same reinforcer rate. With both delayed versus immediate reinforcement and variable-ratio versus variable-interval reinforcement, differential resistance to change is negatively correlated with the log ratios of baseline response rates when reinforcer rates are equated. Cases where resistance to change varies despite equated reinforcer rates, and the correlations among behavioral measures, provide challenges and opportunities for both MPR and BMT. PMID- 12729970 TI - Extending mathematical principles of reinforcement into the domain of behavioral pharmacology. AB - Mathematical principles of reinforcement (MPR) attempts to integrate the empirical laws of reinforcement schedules that have accumulated over the decades. MPR is based on three principles: incentives excite behavior; there are temporal constraints on responding; and coupling of responses to reinforcers strengthens behavior [Behav. Brain Sci. 17 (1994) 105]. In the present paper MPR is extended into the domain of behavioral pharmacology, specifically to model the effects of D-amphetamine on operant behavior. In Experiment 1a, a five-component multiple fixed-ratio schedule was designed to generate behavioral baselines that were subsequently used to assess drug effect. In Experiments 1b and 1c, the quality and quantity of reinforcer were manipulated. The data generated by the three experiments were consistent with MPR. In Experiment 2, MPR was used to model the effects of D-amphetamine on rats responding under the five-component multiple fixed-ratio schedule. According to the model, the rate-decreasing effects of D amphetamine were due primarily to motor disruption and secondarily to increased impulsivity; at the highest dosages, D-amphetamine also may have decreased the incentive value of food. PMID- 12729971 TI - Conditioning from an information processing perspective. AB - The framework provided by Claude Shannon's [Bell Syst. Technol. J. 27 (1948) 623] theory of information leads to a quantitatively oriented reconceptualization of the processes that mediate conditioning. The focus shifts from processes set in motion by individual events to processes sensitive to the information carried by the flow of events. The conception of what properties of the conditioned and unconditioned stimuli are important shifts from the tangible properties to the intangible properties of number, duration, frequency and contingency. In this view, a stimulus becomes a CS if its onset substantially reduces the subject's uncertainty about the time of occurrence of the next US. One way to represent the subject's knowledge of that time of occurrence is by the cumulative probability function, which has two limiting forms: (1) The state of maximal uncertainty (minimal knowledge) is represented by the inverse exponential function for the random rate condition, in which the US is equally likely at any moment. (2) The limit to the subject's attainable certainty is represented by the cumulative normal function, whose momentary expectation is the CS-US latency minus the time elapsed since CS onset. Its standard deviation is the Weber fraction times the CS US latency. PMID- 12729972 TI - Stepping outside the box in considering the C/T ratio. AB - Rate expectancy theory (RET) predicts that in Pavlovian procedures conditioned responding will be directly related to the ratio of time spent in the experimental context (C) relative to the trial time (T) or duration of the conditioned stimulus (CS). This prediction was discussed in the context of three experiments. The first and third experiments involved sexual conditioning in quail [Learn Motiv 31 (2000) 211; J Exp Psychol Anim Behav Processes 27 (2001) 269]; the second experiment involved conditioning rats with food as the unconditioned stimulus (US) [Learn Motiv 28(1997) 465]. In each experiment, one type of conditioned response was directly related to the C/T ratio (as predicted by RET) but another conditioned response was inversely related. In addition, the conditioned behavior that occurred with low C/T ratios was controlled by contextual cues rather than the putative CS. The implications of these findings for possible boundary conditions of RET were discussed. The discussion revealed serious shortcomings in the characterization of context conditioning by RET. PMID- 12729973 TI - Challenges to timing-based theories of operant behavior. AB - Recent theories of behavior have proposed that associative learning principles be replaced by a theoretical framework that assumes the animal has a veridical record of the temporal relations between events. I argue here that such a theory omits critical features of learned behavior: functional differences between different types of temporal relations, the critical nature of response-reinforcer delays, and the necessity of conditioned value as a theoretical construct. PMID- 12729974 TI - Successive two-item same-different discrimination and concept learning by pigeons. AB - Four pigeons were trained in a successive same/different procedure involving the alternation of two stimuli per trial. Using a go/no-go procedure, two different or two identical color photographs were alternated, with a brief, dark, inter stimulus interval, on a computer screen for 20s. Pigeons learned to discriminate between same (S+) and different (D-) sequences with moderate to large contrasts between successive pictures. Analyses of pecking behavior within single trials revealed this discrimination emerged at the earliest possible point in the sequence (i.e. by the presentation of the second item). Pigeons transferred to novel color and gray-scale pictures, and showed savings in tests with novel video stimuli. These results suggest that same/different discrimination and concept formation can be acquired with successively presented pairs of stimuli by pigeons. When combined with results using simultaneous same/different presentations, these findings further support a qualitative similarity among birds and primates in their capacity to judge certain types of stimulus relations. PMID- 12729975 TI - Toward a theory of variability discrimination: finding differences. AB - We sketch the outlines of a theory of variability discrimination that aggregates localized differences to mediate variability discrimination. This Finding Differences Model was compared to a Positional Entropy Model across four different data sets. Although the two models provide strong and similar fits across three of the data sets, only the Finding Differences Model is applicable to investigations involving multidimensional variability. Furthermore, the Finding Differences Model is based on an activation map that has been shown to have utility for visual search tasks, thus establishing its generality across task domains. PMID- 12729976 TI - Temporal discrimination in a long operant chamber. AB - Pigeons were placed in a long chamber equipped with one key and feeder at each end side and one key and houselight at the middle. To obtain food the birds had to choose one side key after a short signal and the other side key after a long signal. The signals consisted of the illumination of the center key and the houselight and were initiated by a peck at the center key. The chamber had sensitive floor panels that enabled us to measure the location of the bird during the signals. In Experiment 1, after the birds learned the discrimination we reversed the assignment of keys to signals. In Experiment 2, we examined performance on two pairs of discriminations holding the same ratio. In Experiment 3, after the pigeons learned to discriminate two signals, we changed the duration of the long signal. The results showed that (a) the birds' motion during the signal was highly stereotypical, i.e. the birds moved to the short side, waited a few seconds, and then departed to, and stayed on the long side; (b) this motion pattern predicted the results of generalization tests with novel durations; (c) the mean of the times of departure from the short side approached its steady state values quicker than the standard deviation and consequently superposition of behavioral measures became stronger with training; (d) only the duration of the short signal influenced significantly the moment the birds departed from the short side; finally (e) the times of arrival at and departure from the short side were positively correlated, but the times of arrival and residence at the short side were negatively correlated. PMID- 12729977 TI - Theoretical note: simulating latent inhibition with selection neural networks. AB - The selection neural-network model proposed by Donahoe et al. [J. Exp. Anal. Behav. 60 (1993) 17] was used to simulate latent inhibition (LI). The model can simulate increases of LI by the number, intensity, and duration of preexposed conditioned stimulus (CS). It can also simulate dependence on total CS preexposure time, CS specificity, and attenuation by preexposure to a compound that includes the to-be-trained CS. It also predicts a potentially new phenomenon: acquisition facilitation by preexposure to a stimulus that is orthogonal to and synaptically competitive with the to-be-trained CS. The basic mechanism is the same through which the model simulates extinction, namely, weight decrement. The realization of this mechanism in the present simulations required two conditions. First, networks had to come to the experimental situation with substantial initial connection weights in the sensory-association subnetwork (0.15, compared to the 0.01 value we have used in all previous simulations). Second, the discrepancy threshold for deciding whether to increase or decrease weights had to be larger than zero (the value we have used in all published simulations). A value of 0.001 was sufficient to produce all the effects. PMID- 12729978 TI - Theories in Progress. PMID- 12729979 TI - Focusing on focus groups. PMID- 12729980 TI - Adolescents' acquisition of cigarettes through noncommercial sources. PMID- 12729981 TI - Hazing of suburban middle school and high school athletes. AB - Little is known about the prevalence of hazing behaviors among adolescent athletes. Our survey of 1105 6th-12th grade athletes found that 17.4% had been subjected to practices that qualify as hazing. Many of these teenagers do not comprehend the abusive and potentially dangerous nature of hazing. PMID- 12729982 TI - Screening for sexually transmitted diseases during preparticipation sports examination of high school adolescents. AB - In an urban school district, 636 students in grades 9-12 were tested for Chlamydia trachomatis and Neisseria gonorrhoeae by ligase chain reaction assays using specimens collected for routine urinalyses during sports physical examinations. Chlamydia and gonorrhea prevalences were 2.8% and 0.7% among males, and 6.5% and 2.0% among females, respectively. Among athletes infected with either sexually transmitted disease (STD), 93.1% reported no symptoms, and treatment was documented for 75.9%. Sports physicals offered a unique opportunity to screen and treat adolescents for STDs and to provide STD-prevention counseling. PMID- 12729983 TI - Dual method use in adolescents: a review and framework for research on use of STD and pregnancy protection. PMID- 12729984 TI - Adolescent substance use, sexual behavior, and metropolitan status: is "urban" a risk factor? AB - PURPOSE: To determine if urban youth ("metropolitan" status) are at greater risk of engaging in risk behaviors than suburban or rural youth. METHODS: We analyzed data on substance use and sexual risk behaviors from the national school-based Youth Risk Behavior Survey (YRBS) conducted in 1999, an anonymous questionnaire self-administered by students in grades 9 through 12. The national survey employs a multistage cluster sample to produce a nationally representative sample of high school students. Data were analyzed using SUDAAN software to take into account the sampling model. RESULTS: In 1999, metropolitan status was not a significant determining factor for involvement in risk behaviors. Of the specific risk factors examined in this analysis, there were no significant differences between rural and suburban youth, and these two groups were combined as "nonurban." In subsequent analysis of urban vs. nonurban youth, no significant differences in risk behaviors were found on bivariate or multivariate analyses. CONCLUSIONS: This analysis suggests that metropolitan status has little if any association with youth engaging in substance use and sexual risk behaviors. In addition, it appears that urban youth are engaging in these risk behaviors no more frequently than their nonurban counterparts. PMID- 12729985 TI - Episodic and persistent gun-carrying among urban African-American adolescents. AB - PURPOSE: To examine whether similar risk factors influenced episodic and persistent gun-carrying among urban African-American adolescents. METHODS: The sample consisted of 705 African-American youths (48.9% male; mean age at baseline = 14.56 years) who were interviewed annually throughout high school as part of a larger study on students who leave school before graduation. Episodic gun carrying was defined as carrying a gun during one or two waves of the study. Persistent gun-carrying involved carrying a gun during three or four waves. Data were analyzed using multinomial logistic regression to test how risk factors assessed at ninth grade influenced the persistence of gun-carrying. RESULTS: Fifteen percent of students reported carrying a gun episodically, and 5% persistently. "Male gender" (OR = 3.61, 95% CI = 2.16-6.04), "adult weapon carrying" (OR = 1.58, 95% CI = 1.20-2.09), "marijuana use" (OR = 1.03, 95% CI = 1.01-1.06), "selling drugs" (OR = 3.24, 95% CI = 1.52-6.92), and "fighting" (OR = 1.57, 95% CI = 1.14-2.15) distinguished noncarriers from episodic carriers. Frequency of fighting (OR = 1.60, 95% CI = 1.00-2.57) and selling drugs (OR = 3.29, 95% CI = 1.16-9.35) distinguished episodic gun-carriers from persistent gun carriers. Variables associated with victimization did not uniquely differentiate among the patterns of gun-carrying. These results were similar for males and females. CONCLUSIONS: Similar risk factors characterize episodic and persistent gun-carrying. Specifically, selling drugs and fighting had a strong dose-response relationship with the persistence of gun-carrying. In this population, episodic gun-carrying should be viewed as very risky and not merely as youthful experimentation or a defensive behavior. PMID- 12729986 TI - Influences on adolescent eating patterns: the importance of family meals. AB - PURPOSE: To provide national estimates of the frequency and determinants of adolescents' consumption of fruits, vegetables, and dairy foods. METHODS: Analyses were based on 18,177 adolescents in the first interview of the National Longitudinal Study of Adolescent Health. Multivariate logistic regressions provide estimates of the unique contribution of sociodemographic characteristics, body weight perception, and parental influences on adolescent food consumption. RESULTS: Almost one in five adolescents reported skipping breakfast the previous day. A large percentage of adolescents reported eating less than the recommended amount of vegetables (71%), fruits (55%), and dairy foods (47%). Adolescents with better-educated parents had better consumption patterns than those with less educated parents. Consumption patterns differed significantly by race. Adolescents who perceived themselves to be overweight were significantly more likely to have poor consumption patterns. Parental presence at the evening meal was associated with a lower risk of poor consumption of fruits, vegetables, and diary foods as well as the likelihood of skipping breakfast. CONCLUSION: Parental presence at the evening meal is positively associated with adolescents' higher consumption of fruits, vegetables, and dairy foods. Nutrition and health professionals should educate parents about the role of family mealtimes for healthy adolescent nutrition. PMID- 12729987 TI - Health-compromising behaviors among Vietnamese adolescents: the role of education and extracurricular activities. AB - PURPOSE: To examine the prevalence of unhealthy behaviors among a cohort of Vietnamese adolescents in California; to examine the relationship between these behaviors and school-related variables (school performance, educational risk behaviors, higher-education aspirations, and participation in extracurricular activities); and to assess the differences that may exist between males and females with regard to these factors. METHODS: We conducted telephone interviews with 783 Vietnamese adolescents, aged 12-17 years, recruited through telephone listings from four California counties where large Vietnamese populations reside: San Francisco, Santa Clara, Los Angeles, and Orange. Of the 783 completed interviews, 60.8% were conducted in English and 39.2% in Vietnamese. The main outcome measure is a health risk behavior scale that includes adolescents' reports of ever smoking a cigarette, sedentary vs. active lifestyle, consumption of fruits and vegetables, consumption of foods high in fat, ever drinking alcohol, and ever engaging in sexual behavior. Multiple regression analyses were employed to estimate the association among the demographic variables, acculturation, school performance, aspirations, extracurricular activities, and the overall health risk. RESULTS: Females were significantly more sedentary than males. Over one-quarter (29%) of the females reported not having participated in vigorous physical activity on 3 or more days per week, compared with just 18% of the males. Most adolescents reported they had never tried cigarettes (84%), never used alcohol (77%), and never had sex (97%). Males were more likely than females to report a higher frequency of experimentation with smoking and drinking. Overall, school performance and participation in extracurricular activities were significantly related to the health risk behavior scale. Adolescents who demonstrated at least one educational risk (ever skipped school or ever sent out of the classroom) were more likely to engage in other risky behaviors. Also, older and more acculturated adolescents were at increased risk of engaging in health-compromising behaviors. Analysis by gender revealed that the variables age, educational risk, and chance of attending college were all related to health risk behavior for both males and females. Among the boys, those who reported achieving an average grade of B or better had a decreased risk of engaging in health-compromising behaviors; however, neither extracurricular activities nor acculturation was related to health-compromising behaviors in boys. Among the girls, the reverse was true: lack of participation in extracurricular activities was related to health-compromising behaviors, whereas grades were not a significant risk factor. CONCLUSIONS: Among sampled Vietnamese adolescents in California, health risk behaviors are common and inversely related to some school performance indicators. Using these indicators to identify high-risk groups could allow targeted educational programs or interventions for the mitigation of health compromising behaviors. PMID- 12729988 TI - Corporal punishment in schools: position paper of the Society for Adolescent Medicine. PMID- 12729989 TI - Confirmation of extended-spectrum beta-lactamase-producing Serratia marcescens: preliminary report from Taiwan. AB - Although Serratia marcescens is a common cause of nosocomial infection in Taiwan, strains producing extended-spectrum beta-lactamases (ESBLs) are rare. We detected four clinical isolates of S. marcescens from Taiwan that exhibited resistance to cefotaxime (MICs, > 256 microg/ml) and cefepime (MICs, > or = 32 microg/ml), but were susceptible to imipenem and meropenem. Transconjugants revealed similar MIC profiles when compared to the parental strains. Isoelectric focusing revealed one major transferable beta-lactamase (pI 8.4), which was further identified as CTX-M 3 by polymerase chain reaction and gene sequencing. An AmpC-like enzyme (pI 8.8) was not transferable. All four isolates had significant MIC reductions of > or =3 log(2) dilutions for cefepime in the presence of clavulanic acid, compatible with the presence of an ESBL (CTX-M-3). Clavulanate did not significantly reduce the cefotaxime MIC for one isolate that may co-produce high-level AmpC beta-lactamase (pI 8.8). Since high-level AmpC expression has minimal effect on the activity of cefepime, isolates co-producing AmpC beta-lactamase may be recognized as additional ESBL producers by using cefepime as an ESBL screening agent. PMID- 12729990 TI - Cross-reactive monoclonal antibodies raised against the lipopolysaccharide antigen of Salmonella minnesota Re chemotype: diagnostic relevance. AB - Three cross-reactive monoclonal antibodies (MAbs) of IgM and IgG2b isotype were generated in two separate fusions after immunization of BALB/c mice with heat killed Salmonella minnesota R595 of Re chemotype and acid-treated bacteria, coated with Re lipopolysaccharide (LPS) antigen. The specificity of the MAbs was demonstrated as the Re LPS antigen. The activity and cross-reactivity against purified elementary bodies of Chlamydia trachomatis and various S- and R-LPS antigens of other Gram-negative bacteria were characterized in enzyme-linked immunosorbent assay, passive hemolysis assay, immunoblot and immunofluorescence with the available chlamydial strains. The results demonstrated cross-reaction between the Re LPS antigen, the genus-specific chlamydial LPS and the LPS antigens of Escherichia coli O119 and Acinetobacter baumannii, suggesting the presence of identical or similar epitopes in the lipopolysaccharide antigens. The findings are implying the necessity of novel approaches, improving the specificity of serologic assays in the laboratory diagnosis of chlamydial infections. PMID- 12729992 TI - Use of the NOW Streptococcus pneumoniae urinary antigen test in cerebrospinal fluid for rapid diagnosis of pneumococcal meningitis. AB - Streptococcus pneumoniae is one of the most common pathogens in bacterial meningitis. Rapid diagnosis is critical for effective treatment. The aim of this study was to assess the accuracy of the NOW S. pneumoniae Urinary Antigen Test, (Binax, Portland, ME, USA) originally developed for urine testing, in detecting the S. pneumoniae antigen in cerebrospinal fluid (CSF). The study included 519 patients with suspected meningitis. CSF, blood and urine samples were cultured according to standard methods. CSF viral culture was also performed. CSF and urine specimens were tested for pneumococcal antigen with the NOW S. pneumoniae test.S. pneumoniae was isolated from the CSF of 22 patients. The direct antigen test was positive in CSF in 21/22 patients (95.4% sensitivity), and in urine, in 12/21 (57.1% sensitivity). Direct CSF smear was positive in 15/22 (68% sensitivity). CSF samples that cultured negative for S. pneumoniae (n = 470) or positive for other bacteria (n = 27) were also negative on the NOW test (100% specificity). By contrast, urine samples of 63/470 of patients with negative CSF culture were positive on the NOW test, as were 5/27 urine samples of patients with CSF culture positive for other bacteria (p = 0.45). The NOW S. pneumoniae antigen test in CSF yields a rapid and very reliable diagnosis of pneumococcal meningitis, enabling prompt and adequate treatment. Its low sensitivity in urine indicates that this mode of testing is not useful for the diagnosis of pneumococcal meningitis. These data have been included in the FDA application for approval of the NOW test for use in the CSF for the diagnosis of pneumococcal meningitis. PMID- 12729993 TI - In vitro susceptibility testing of filamentous fungi: comparison of Etest and reference M38-A microdilution methods for determining posaconazole MICs. AB - The performance of the Etest for posaconazole susceptibility testing of 72 isolates of filamentous fungi was assessed in comparison with the National Committee for Clinical Laboratory Standards (NCCLS) approved standard broth microdilution method (M38-A). The NCCLS method employed RPMI 1640 broth medium, and MICs were read after incubation for 48h at 35 degrees C. Etest MICs were determined with RPMI agar containing 2% glucose and were read after incubation for 48h at 35 degrees C. The isolates included Aspergillus fumigatus, A. flavus, A. nidulans, A. niger, A. versicolor, A. oryzae, A. terreus, Cladosporium spp., Curvularia sp., Exophiala sp., Fusarium spp., Paecilomyces spp., Pithomyces sp., Penicillium spp. and Scedosporium apiospermum. Overall agreement between Etest and microdilution MICs was 84% for Aspergillus spp. and 100% for the less common opportunistic molds, with the exception of Penicillium spp. (67%). Where a discrepancy was observed between Etest and the reference method, the Etest tended to give lower values. The Etest method using RPMI agar appears to be a useful method for determining posaconazole susceptibilities of filamentous fungi. PMID- 12729994 TI - Phenotypic antimicrobial resistance patterns in Pseudomonas aeruginosa and Acinetobacter: results of a Multicenter Intensive Care Unit Surveillance Study, 1995-2000. AB - Susceptibility data from the Intensive Care Unit (ICU) Surveillance Study for 10,361 isolates of Pseudomonas aeruginosa and 2,573 isolates of Acinetobacter tested at centers in the United States during 1995 to 2000 were analyzed. In all years, amikacin was the most active antimicrobial agent against P. aeruginosa, and imipenem was the most active agent against Acinetobacter. Resistance of both organisms to common therapeutic agents tested throughout the analysis period increased from 1995 to 2000, although the increase was not consistent for all drugs from year to year. The increases were higher among Acinetobacter, and for both organisms, the increase in resistance was greatest for ciprofloxacin. Among all P. aeruginosa tested in 1999 and 2000, resistance to ciprofloxacin was 9-11% higher for isolates from patients on general hospital wards than those from ICUs. Of the 3424 ICU isolates of P. aeruginosa tested in 1999 and 2000, 77 (2.2%) were multidrug-resistant (i.e., resistant to piperacillin, ceftazidime, imipenem, and gentamicin). Twenty (3.9%) isolates of Acinetobacter in 1999 and seven (1.9%) in 2000 were resistant to imipenem, ceftazidime, piperacillin-tazobactam, ciprofloxacin, and amikacin. Although resistance in both P. aeruginosa and Acinetobacter increased, multidrug-resistant (to > or =4 agents) strains were uncommon. PMID- 12729991 TI - Circulating levels of FAS/APO-1 in patients with the systemic inflammatory response syndrome. AB - Resolution of inflammation/infection involves removal of neutrophils and other inflammatory cells by the induction of apoptosis. Fas/Apo-1 is a widely occurring apoptotic signal receptor molecule expressed by almost any type of cell, which is also released in a soluble circulating form. In this study we investigated the role of circulating Fas/Apo-1 in patients with systemic inflammatory response syndrome (SIRS). We evaluated 57 critically ill patients, 34 with infectious SIRS (sepsis and septic shock), and 23 patients with noninfectious SIRS. Circulating Fas/Apo-1 was determined by a commercially available immunoassay. Our results clearly show that levels of Fas/Apo-1 were significantly elevated in patients with infectious and noninfectious SIRS (10.4 +/- 8.1 pg/mL, controls: 5.0 +/- 0.7 pg/mL; p < 0.0001). In addition, Fas/Apo-1 levels were not able in predicting in predicting poor outcome of patients with SIRS. In conclusion, these results show that increased levels of Fas/Apo-1 from patients with SIRS is a mechanism which contribute to inflammatory response through accumulation of neutrophils at sites of inflammation/infection. PMID- 12729996 TI - Identification of an epidemic carbapenem-resistant Acinetobacter baumannii strain at hospitals in Buenos Aires City. AB - To identify epidemic Acinetobacter baumannii (AB) clones, 38 carbapenem-resistant AB isolates from 5 hospitals were analyzed. Macrorestriction classified 24 isolates as clone IV, susceptibility pattern clustering analysis grouped almost all of them together, and they were uniformly biotype 8. Clone IV was present at all 5 hospitals, so that it represents a carbapenem-resistant AB strain with epidemic behavior. PMID- 12729995 TI - Antimicrobial susceptibility of community-acquired respiratory tract pathogens in North America in 1999-2000: findings of the PROTEKT surveillance study. AB - The PROTEKT surveillance study commenced in 1999 to examine the antimicrobial susceptibility of community-acquired respiratory pathogens. We report here the results from 2371 isolates collected during 2000 by North American centers (Canada, n = 7; USA, n = 8). Overall, 21.3% of pneumococci (n = 687) were penicillin G-resistant (Canada, 10.3%; USA, 32.6%). Corresponding rates of erythromycin resistance were 16.3% and 31.5%. Telithromycin inhibited all penicillin- and erythromycin-resistant isolates at < or =1 microg/ml. Among 612 Hemophilus influenzae isolates, 22.4% were beta-lactamase-positive. Antimicrobial susceptibility of H. influenzae varied between 82.4% (clarithromycin) and 100% (cefpodoxime, levofloxacin). Importantly, one isolate was found to be resistant to both moxifloxacin and ciprofloxacin. Most Moraxella catarrhalis isolates were highly susceptible to the antimicrobials tested, except ampicillin. All Streptococcus pyogenes isolates (n = 382) were penicillin-susceptible and 5.2% were non-susceptible to erythromycin. S. pyogenes showed cross-resistance to other macrolides yet remained inhibited by telithromycin at < or =0.5 microg/ml. Methicillin resistance among Staphylococcus aureus was common (19.9%), particularly in the USA. The PROTEKT study confirms the widespread prevalence of antimicrobial resistance among common respiratory pathogens in North America, and hence the need for continued surveillance to guide optimal empiric therapy for community-acquired respiratory tract infections. PMID- 12729997 TI - Mutation prevention concentration of ceftriaxone, meropenem, imipenem, and ertapenem against three strains of Streptococcus pneumoniae. AB - This investigation tested the mutation prevention concentration (MPC) concept using imipenem, meropenem, ceftriaxone, and ertapenem against three strains of Streptococcus pneumoniae (PCN MIC = 0.012, 1, 8 mg/L, respectively). MIC, MBC, and MPC values for each of the beta-lactams did not differ by more than one tube dilution. While an interesting concept, MPC may not apply to antimicrobials that do not utilize a dual targeting system, such as beta-lactams, or to bacteria that exhibit multiple mechanisms of resistance and/or mutate at a rate where the frequency would likely be captured by the standard inoculum size used in routine MIC testing. PMID- 12729998 TI - Comparison of real-time PCR detection methods for B1 and P30 genes of Toxoplasma gondii. AB - Real-time PCR assays were established with the Toxoplasma gondii P30 and the 35 copy B1 gene as target genes. Both PCR methods detected Toxoplasma DNA from 10 to 100000 genome equivalents per assay and had comparable intra-assay deviations suggesting that detecting a single copy locus is suitable for clinical diagnostic. PMID- 12729999 TI - Comparative activity of garenoxacin (BMS 284756), a novel desfluoroquinolone, tested against 8,331 isolates from community-acquired respiratory tract infections: North American results from the SENTRY Antimicrobial Surveillance Program (1999-2001). AB - Emerging resistances to orally administered antimicrobials have escalated among bacteria causing community-acquired respiratory infections (CARTI). The spectrum and potency of garenoxacin, (formerly BMS 284756) was assessed against a collection of CARTI isolates from North American medical centers during a longitudinal surveillance study, the SENTRY Antimicrobial Surveillance Program (1999-2001). A total of 8,331 strains of Hemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae were tested by reference methods and compared to numerous other marketed antimicrobials. Nearly all (95.1%) M. catarrhalis were penicillin-resistant, 27.9% of H. influenzae were ampicillin resistant and 35.0% of S. pneumoniae had MICs at > or = 0.12 microg/ml for penicillin. Garenoxacin was very active against the three monitored species with MIC(90) values of < or =0.06 microg/ml. Garenoxacin and other quinolones were equally active against the Gram-negative pathogens (except moxifloxacin which was at least twofold less potent versus M. catarrhalis). However, against pneumococci the rank order of potency (MIC(50) in microg/ml) was: gemifloxacin (0.015) > garenoxacin (0.06) > trovafloxacin = moxifloxacin (0.12) > gatifloxacin (0.25) > levofloxacin = ciprofloxacin (1). A trend toward greater resistance worldwide was observed for ciprofloxacin (MIC, > or = 4 microg/ml), increasing from 1.5% in 1999 to 6.8% in 2001. The highest quinolone resistance rate was observed in North America. Garenoxacin, a new desfluoro(6)quinolone, was documented to be very active in vitro (MIC, < or =2 microg/ml) against > 99.9% of all CARTI isolates in the SENTRY Program. Evolving resistances to other antimicrobial classes or among currently used quinolones appear to position this investigational desfluoro(6)quinolone as a potential treatment option for future clinical use in ambulatory patients. PMID- 12730000 TI - Pathogen of occurrence and susceptibility patterns associated with pneumonia in hospitalized patients in North America: results of the SENTRY Antimicrobial Surveillance Study (2000). AB - Antimicrobial selection for patients diagnosed with pneumonia is a major therapeutic challenge and dilemma to the clinical practitioner. In the community setting, patients usually receive empiric oral therapy based upon multiple patient risk factors and locally prevalent pathogen susceptibilities. For patients admitted to the hospital with pneumonia, or who acquire pneumonia while in the hospital, therapy can be initially empiric and then become directed once culture and susceptibility results are known. The SENTRY Antimicrobial Surveillance Program since 1997, has monitored pathogen frequency and antimicrobial susceptibilities in hospitalized patients with pneumonia in North America. In this Study 2,712 pathogens were studied from 30 medical centers (25 in the United States and 5 in Canada). Over 30 species of organisms were recovered with Staphylococcus aureus comprising 28.0% of all isolates and with four other species (Pseudomonas aeruginosa 20.0%, Streptococcus pneumoniae 9.1%, Klebsiella spp. 7.5% and Haemophilus influenzae 7.3%) constituted 71.9% of isolates submitted. Methicillin (oxacillin)-resistant S. aureus accounted for 43.7% of all S. aureus isolates. Antimicrobials demonstrating significant (>80%) activity against S. aureus were: chloramphenicol (81.6%), tetracycline (91.4%), rifampin (96.4%) and quinupristin/dalfopristin (99.7%); and no isolate was resistant to glycopeptides or linezolid. North American isolates of P. aeruginosa were most susceptible to amikacin (93.7%) > tobramycin (90.2%) > meropenem (89.1%) > imipenem = piperacillin/tazobactam (85.6%) > piperacillin (82.1%) > cefepime (80.5%). Overall, 32.1% of S. pneumoniae were penicillin non-susceptible while erythromycin susceptibility was only 74.8%. Fluoroquinolones and recent generation cephalosporins retained excellent activity (gatifloxacin [99.2%] > levofloxacin = cefepime [98.8%] > ceftriaxone [97.2%]) against S. pneumoniae. Klebsiella spp. were 100.0% susceptible to the carbapenems (meropenem and imipenem) but extended spectrum beta-lactamases were detected at a rate of 5.4%. The beta-lactamase-positive rate in H. influenzae was 28.6% in North America (71.4% ampicillin-susceptible). The SENTRY Antimicrobial Surveillance Program continues to identify important North American patterns of pathogen frequency and resistance. Additionally, the provision of multi-year longitudinal data and associated reports allow for comparisons, which function as critical tools for effective patient management and antimicrobial interventions. PMID- 12730001 TI - Occurrence and antimicrobial susceptibility patterns of pathogens isolated from skin and soft tissue infections: report from the SENTRY Antimicrobial Surveillance Program (United States and Canada, 2000). AB - A total of 1,404 bacterial isolates were recovered from skin and soft tissue infections (SSTIs) from hospitalized patients in 24 sites in the United States (US) and 5 Canadian medical centers as part of the SENTRY Antimicrobial Surveillance Program. Isolates were collected between October and December, 2000. The rank order of pathogens was: Staphylococcus aureus (45.9%), Pseudomonas aeruginosa (10.8%), Enterococcus spp. (8.2%), Escherichia coli (7.0%), Enterobacter spp. (5.8%) and Klebsiella spp. (5.1%). The same order was observed in the US and Canada. Of note, almost 30% of S. aureus were oxacillin-resistant. Vancomycin resistance among enterococci was low (7.8%) representing a marked decrease from earlier SENTRY Program reports. Several antimicrobial agents remained very active against P. aeruginosa and Enterobacteriaceae isolates. In particular amikacin, cefepime, and the carbapenems (imipenem and meropenem) showed an excellent spectrum of activity (>95% susceptible). Extended-spectrum beta-lactamase production was observed in both E. coli (7.1%) and Klebsiella spp. (11.3%). Cefepime remained highly active, even against ceftazidime-resistant isolates of Enterobacter spp. The results of this study have identified the most common causes of SSTIs in hospitalized patients in North America, and can be used to make informed decisions concerning standards of empiric treatment for SSTIs in this region. PMID- 12730002 TI - Susceptibility patterns of orally administered antimicrobials among urinary tract infection pathogens from hospitalized patients in North America: comparison report to Europe and Latin America. Results from the SENTRY Antimicrobial Surveillance Program (2000). AB - Urinary tract infections (UTIs) remain a worldwide nosocomial infection problem. Geographic variations in pathogen occurrence and susceptibility profiles require monitoring to provide information to guide new (garenoxacin [BMS284756]) therapeutic options. Two thousand seven hundred-eighty UTI isolates from Europe (n = 783), Latin America (531), and North America (1,466) were tested and compared against 44 agents by reference methods in the SENTRY Antimicrobial Surveillance Program. The top seven pathogens accounted for 90% of all isolates and the rank order for all regions was: Escherichia coli (1,316; 47%), Enterococcus spp. (351; 13%), Klebsiella spp. (306; 11%), Pseudomonas aeruginosa (210; 8%), Proteus mirabilis (145; 5%), Enterobacter spp. (97; 4%), and Citrobacter spp. (78; 3%). The pathogen rank order was similar among regions except for the rarer occurrence of Enterococcus spp. (Rank #6, 4%) in Latin America. E. coli ampicillin resistance was highest in Europe and Latin America (51-55%). Ampicillin (37%), ciprofloxacin or garenoxacin (4%), and trimethoprim/sulfamethoxazole (23%) resistance remained lowest in North America. Nitrofurantoin susceptibility in E. coli was still at acceptable levels and ranged from 91 to 96% across regions. The regional ciprofloxacin-resistant rank order for P. aeruginosa by region was: Latin America (55%) > Europe (41%) > North America (29%). Vancomycin-resistant enterococci (VRE) were only detected in North America (7%). Garenoxacin possessed a 34 to 44% wider spectrum compared to ciprofloxacin against enterococci UTI isolates. Extended spectrum beta-lactamase rates for E. coli and Klebsiella spp. were 4 and 19%, respectively. These results emphasized the need to assess the often striking differences in pathogen occurrence and resistance rates among the commonly encountered UTI pathogens. PMID- 12730003 TI - Assessing viability and cell-associated product of recombinant protein producing Pichia pastoris with flow cytometry. AB - This paper describes the establishment of flow cytometric methods for recombinant Pichia pastoris strains, and their application to a lab scale fed batch fermentation. Using a strain which secretes human trypsinogen, the viability and the product which remained associated to the cell were measured with propidium iodide and immunofluorescent staining, respectively. Viability decreases significantly below 70% during the methanol fed batch phase, indicating a stress situation triggered by the fermentation conditions. Cell associated product is accumulated earlier after methanol induction than secreted product. These data demonstrate that flow cytometry is a powerful tool for the analysis and optimization of recombinant protein production processes, and they indicate the need to further improve a widely used fermentation protocol for P. pastoris. PMID- 12730004 TI - Effect of hyperosmolarity on recombinant protein productivity in baculovirus expression system. AB - A lot of strategies were applied to improve recombinant protein productivity in the baculovirus expression system. In this study we propose for foreign protein production fed-batch cultivation method at hyperosmotic environment induced by increased NaCl content. Obtained results suggested relatively high tolerance and adaptation abilities of Tn-5 insect cells exposed to hyperosmotic stress. The cells under hyperosmotic conditions increased the specific rate of glucose consumption and lactate production. The release of additional energy and precursors as a result of increased metabolism by osmotically stressed culture was involved in recombinant protein synthesis. Recombinant nucleoprotein productivity in nutritional feeding cultures exposed to hyperosmolarity was about 72% higher than that obtained in batch culture at physiological osmolarity, but 31% was a result of feeding and the rest 41% was a result of hyperosmolarity and increasing Na(+) concentration. PMID- 12730005 TI - Construction of a bacterial biosensor for styrene. AB - A new bacterial biosensor for styrene has been developed and characterized. A translational fusion of the lacZ gene to the sty promoter of Pseudomonas sp. strain Y2 has been inserted into miniTn5. Transposition of the recombinant transposon to the chromosome of Pseudomonas sp. strain Y2 resulted in a whole cell biosensor able to detect and degrade styrene. In this biosensor, the endogenous StyS/StyR system detects the presence of styrene and turns on the expression of the exogenous reporter gene from the transferred construction. Other compounds such as toluene, epoxystyrene, phenylacetaldehyde and 2 phenylethanol also induced expression of beta-galactosidase although quantitative differences in their effect are clearly detected. Non-inducing compounds affect differently the sensitivity to inducing compounds when present in a mixture. PMID- 12730006 TI - A general model for biosorption of Cd2+, Cu2+ and Zn2+ by aerobic granules. AB - Aerobic granules are microbial aggregates with a strong and compact structure. This study looked into the feasibility of aerobic granules as a novel type of biosorbent for the removal of individual Cd(2+), Cu(2+) and Zn(2+) from aqueous solution. Based on the thermodynamics of biosorption reaction, a general model was developed to describe the equilibrium biosorption of individual Cd(2+), Cu(2+) and Zn(2+) by aerobic granules. This model provides good insights into the thermodynamic mechanisms of biosorption of heavy metals. The model prediction was in good agreement with the experimental data obtained. It was further demonstrated that the Langmuir, Freundlich and Sips or Hill equations were particular cases of the proposed model. The biosorption capacity of individual Cd(2+), Cu(2+) and Zn(2+) on aerobic granules was 172.7, 59.6 and 164.5 mgg(-1), respectively. These values may imply that aerobic granules are effective biosorbent for the removal of Cd(2+), Cu(2+) and Zn(2+) from industrial wastewater. PMID- 12730007 TI - Activity, conformation and dynamics of cutinase adsorbed on poly(methyl methacrylate) latex particles. AB - The adsorption of a recombinant cutinase from Fusarium solani pisi onto the surface of 100 nm diameter poly(methyl methacrylate) (PMMA) latex particles was evaluated. Adsorption of cutinase is a fast process since more than 70% of protein molecules are adsorbed onto PMMA at time zero of experiment, irrespective of the tested conditions. A Langmuir-type model fitted both protein and enzyme activity isotherms at 25 degrees C. Gamma(max) increased from 1.1 to 1.7 mg m(-2) and U(max) increased from 365 to 982 U m(-2) as the pH was raised from 4.5 to 9.2, respectively. A decrease (up to 50%) in specific activity retention was observed at acidic pH values (pH 4.5 and 5.2) while almost no inactivation (eta(act) congruent with 87-94%) was detected upon adsorption at pH 7.0 and 9.2. Concomitantly, far-UV circular dichroism (CD) spectra evidenced a reduction in the alpha-helical content of adsorbed protein at acidic pH values while at neutral and alkaline pH the secondary structure of adsorbed cutinase was similar to that of native protein. Fluorescence anisotropy decays showed the release of some constraints to the local motion of the Trp69 upon protein adsorption at pH 8.0, probably due to the disruption of the tryptophan-alanine hydrogen bond when the tryptophan interacts with the PMMA surface. Structural data associated with activity measurements at pH 7.0 and 9.2 showed that cutinase adsorbs onto PMMA particles in an end-on orientation with active site exposed to solvent and full integrity of cutinase secondary structure. Hydrophobic interactions are likely the major contribution to the adsorption mechanism at neutral and alkaline pH values, and a higher amount of protein is adsorbed to PMMA particles with increasing temperature at pH 9.2. The maximum adsorption increased from 88 to 140 mg cutinase per g PMMA with temperature raising from 25 to 50 degrees C, at pH 9.2. PMID- 12730008 TI - Lipase-catalyzed synthesis of xylitol monoesters: solvent engineering approach. AB - A solvent engineering strategy was applied to the lipase-catalyzed synthesis of xylitol-oleic acid monoesters. The different esterification degrees for this polyhydroxylated molecule were examined in different organic solvent mixtures. In this context, conditions for high selectivity towards monooleoyl xylitol synthesis were enhanced from 6 mol% in pure n-hexane to 73 mol% in 2-methyl-2 propanol/dimethylsulfoxide (DMSO) 80:20 (v/v). On the contrary, the highest production of di- and trioleoyl xylitol, corresponding to 94 mol%, was achieved in n-hexane. Changes in polarity of the reaction medium and in the molecular interactions between solvents and reactants were correlated with the activity coefficients of products. Based on experimental results and calculated thermodynamic activities, the effect of different binary mixtures of solvents on the selective production of xylitol esters is reported. From this analysis, it is concluded that in the more polar conditions (100% dimethylsulfoxide (DMSO)), the synthesis of xylitol monoesters is favored. However, these conditions are unfavorable in terms of enzyme stability. As an alternative, binary mixtures of solvents were proposed. Each mixture of solvents was characterized in terms of the quantitative polarity parameter E(T)(30) and related with the activity coefficients of xylitol esters. To our knowledge, the characterization of solvent mixtures in terms of this polarity parameter and its relationship with the selectivity of the process has not been previously reported. PMID- 12730009 TI - Electroenzymatic oxidation of veratryl alcohol by lignin peroxidase. AB - This paper reports the formation of veratraldehyde by electroenzymatic oxidation of veratryl alcohol (3,4-dimethoxybenzyl alcohol) hybridizing both electrochemical and enzymatic reactions and using lignin peroxidase. The novel electroenzymatic method was found to be effective for replacement of hydrogen peroxide by an electrochemical reactor, which is essential for enzyme activity of lignin peroxidase. The effects of operating parameters such as enzyme dosage, pH, and electric potential were investigated. Further, the kinetics of veratryl alcohol oxidation in an electrochemical reactor were compared to oxidation when hydrogen peroxide was supplied externally. PMID- 12730010 TI - Transfection of partially purified plasmid DNA for high level transient protein expression in HEK293-EBNA cells. AB - One of the major constraints to performing large-scale transfections of cultured mammalian cells for the transient expression of recombinant proteins is the production of large quantities of purified plasmid DNA. In this report partially purified plasmid DNA was prepared by a method that combines alkaline lysis of E. coli with standard precipitation techniques. The efficiency of calcium phosphate DNA co-precipitate formation with crude DNA was similar to that observed for pure DNA, but precipitate formed with crude DNA also contained RNA. The transfection of adherent and suspension-adapted HEK293-EBNA cells with partially purified pEGFPN1 resulted in levels of transient GFP expression equivalent to those achieved with pure DNA. In addition, the co-transfection of 1-200 ml cultures of suspension-adapted HEK293-EBNA cells with two different plasmids encoding the heavy and light chain genes of anti-human RhD IgG1, respectively, yielded similar IgG titers with pure and partially purified plasmid DNA. Finally, it was observed that suspension-adapted cells were more tolerant to the presence of RNA in the plasmid preparations than were adherent cells. These findings are relevant to the field of DNA transfection, including applications ranging from high-throughput screening to large-scale transient protein expression. PMID- 12730011 TI - Association of lymphopenia with porcine circovirus type 2 induced postweaning multisystemic wasting syndrome (PMWS). AB - The composition of peripheral blood leukocyte populations was studied following experimental PCV2-infection in 3-week-old piglets. Four of 10 PCV2-infected piglets developed clinical and pathological symptoms consistent with postweaning multisystemic wasting syndrome (PMWS) between 14 and 21 days post-inoculation (p.i.), and were characterised as PMWS-affected. Only these four PMWS-affected piglets, but neither the non-symptomatic infected nor control animals, developed a clear leukopenia. Kinetic analysis demonstrated a clear loss of both CD21(+) B and CD3(+) T lymphocytes in the PMWS-affected piglets. By CD3/CD4/CD8 triple labelling, the influence of PCV2 infection on all T cell sub-populations was discernible. A loss of CD3(+)CD4(+)CD8(+) memory/activated Th lymphocytes was particularly notable. However, all T lymphocyte sub-populations CD3(+)CD4(+)CD8(+) memory Th, CD3(+)CD4(+)CD8(-) nai;ve Th, CD3(+)CD4(-)CD8(+) Tc and CD3(+)CD4(-)CD8(-) gammadelta TCR(+) lymphocytes-were susceptible to PCV2 infection-induced lymphopenia. CD3(-)CD4(-)CD8(+) NK cells were also depleted in the PMWS-affected animals, but granulocytes and monocytes were less affected. In conclusion, PCV2 infection induces primarily a lymphopenia, but only in animals which subsequently develop PMWS. The lymphopenia can be identified early p.i., particularly with the B lymphocytes. Memory/activated Th lymphocytes might be affected more than the other T cell sub-populations, but as time progressed a collapse of both T and B cell populations was clear. PMID- 12730012 TI - Serum IgE and IgG responses to food antigens in normal and atopic dogs, and dogs with gastrointestinal disease. AB - In human food allergy, with or without concurrent atopy, there may be significant increases in serum allergen-specific IgE. Serological methods have been tried but are not currently recommended for diagnosis of suspected food allergy in dogs. The aim of this study was to investigate humoral immune responses to food antigens in dogs. Serum IgG and IgE antibodies specific for food antigens were measured by enzyme linked immunosorbent assay (ELISA) using polyclonal anti-dog IgG and IgE reagents. Antigens tested were beef, chicken, pork, lamb, chicken, turkey, white fish, whole egg, wheat, soybean, barley, rice, maize corn, potato, yeast and cow's milk. Three groups were examined: normal dogs, dogs with atopic dermatitis (AD); and dogs with one of four types of gastrointestinal (GI) disease: small intestinal bacterial overgrowth (SIBO), inflammatory bowel disease (IBD), food-responsive disease, and infectious diarrhoea. Statistically significant differences in food-specific antibodies were not detected between the GI subgroups. There were statistically significant differences in the IgE concentration between the normal dogs, and dogs with atopic or GI disease, for all of the antigens tested. There were statistically significant differences in the average IgG concentrations between the normal dogs, and dogs with atopic or GI disease, for all of the antigens tested, except egg and yeast. The relationship of antigen responses for pooled data was analysed using principle component analysis and cluster plots. Some clustering of variables was apparent for both IgE and IgG. For example, all dogs (normal and diseased) made a similar IgG antibody response to chicken and turkey. Compared with other groups, atopic dogs had more food allergen-specific IgE and this would be consistent with a Th(2) humoral response to food antigens. Dogs with GI disease had more food allergen-specific IgG compared with the other groups. This may reflect increased antigen exposure due to increased mucosal permeability which is a recognised feature of canine intestinal disease. PMID- 12730013 TI - Identification and characterization of auto-anti-idiotypic antibodies specific for antibodies against porcine reproductive and respiratory syndrome virus envelope glycoprotein (GP5). AB - Auto-anti-idiotypic antibodies (Aab-2s) were detected from pigs experimentally infected with porcine reproductive and respiratory syndrome virus (PRRSV). The Aab-2s were specific against the idiotypic antibodies (Ab-1s) to the envelope glycoprotein GP5 of PRRSV and were detected from serum samples collected between 21 and 98 days post-infection (DPI). Serological characterization indicated that the Aab-2s recognized the idiotype located within or near the antigen-combining sites of the anti-GP5 antibodies, which was shared by both mouse MAb anti-GP5 and swine polyclonal antibodies. The fact that the Aab-2 inhibited the anti-GP5 antibodies from binding to PRRSV and that they were detected at different time periods in pigs that cleared the infection prior to 98 DPI versus pigs in which virus was detected at 98 DPI suggests that Aab-2 antibodies may play a role in immunity to PRRSV infection. PMID- 12730014 TI - IgE ELISA using antisera derived from epsilon chain antigenic peptides detects allergen-specific IgE in allergic horses. AB - Equine disease with an allergic etiology is common. Environmental antigens most often implicated as allergens in horses include molds, dusty hay, grass pollen, hay dust mites, and insect saliva. Although intradermal testing with allergen is a useful diagnostic tool for some species, skin testing frequently produces false positive results in horses. Allergen deprivation as a diagnostic tool is often impossible and at best it is ineffective at diagnosing the specific allergic reactivity. Synthesis of IgE after exposure to allergen is the instigator of the allergic process. While IgE exerts its effect after binding strongly to mast cell Fc receptors, the presence of free IgE in the serum can be used to quantify and determine the allergen specificity of the allergic disease. A lack of widely available reagents for detection of equine IgE has limited this approach in horses. We have used the nucleotide sequence of equine IgE to prepare a peptide based immunogen to elicit equine epsilon chain-specific antisera. Selection of peptides was based on antigenic attributes of the deduced amino acid sequence of the equine epsilon chain. Six peptides were selected for conjugation to carrier molecules and rabbit immunization. Of these, one peptide elicited antisera that was successfully used in enzyme linked immunosorbant assay (ELISA) to screen horse serum from 64 allergic horses for allergen-specific IgE. Twenty-four of the 64 horses showed positive reactivity to one or more of the following allergens: grass, grain mill dust, mosquito, and horsefly. This study demonstrates the usefulness of peptide-based immunogens for development of antisera to rare or difficult to purify antigens such as IgE. Resultant antisera has great usefulness in diagnostic assays for equine allergy and as a research tool. PMID- 12730015 TI - Canine transmissible venereal tumor cell depletion of B lymphocytes: molecule(s) specifically toxic for B cells. AB - Canine transmissible venereal tumor (CTVT) is an excellent model for investigating the interaction between host immunity and tumor growth. Although CTVT is an allograft, initially the host immune system is unable to destroy the tumor cells, and the tumor grows progressively for about 4-6 months (P phase). After a short stable phase, the tumor undergoes regression (R phase). In this study, CTVT inoculation significantly reduced the proportion of B lymphocytes among all peripheral blood lymphocytes (PBL), but the proportion of B lymphocytes returned to normal after complete removal of CTVT. Following CTVT inoculation, immunoglobulin concentrations decreased gradually, coincident with B lymphocyte decline. Furthermore, CTVT secreted a soluble, heat- and protease K-sensitive cytotoxic molecule(s) that destroyed peripheral blood B lymphocytes (PBBL) but spared other types of immune cells regardless of whether mitogens, such as IL-2 or Con A, were present. The decrease in the proportion and viability of PBBL was caused by a cytotoxic molecule(s) that induced apoptosis. The molecular weight of the CTVT-derived cytotoxic molecule(s) was 30-100kDa. Human, domestic cat, horse and mouse B cells were also sensitive to the substance. PMID- 12730017 TI - Williams syndrome: 15 years of psychological research. AB - Williams syndrome is a rare genetic disorder caused by a microdeletion of about 20 genes on chromosome 7q11.23. The importance of this syndrome for theories and models of cognition became apparent about 15 years ago, when Bellugi, Sabo, and Vaid (1988) argued that Williams syndrome was a paradigm case of the independence of language from cognition. In particular, it was argued that individuals with Williams syndrome have "intact" language despite severe mental retardation. More recent findings indicate a more complex relation between language and characteristics associated with this syndrome. The articles in this special issue form 3 overlapping clusters: articles concerned with language development relative to cognitive development, articles concerned with other aspects of cognition, and articles concerned with interpersonal relations and personality. Together, these articles provide strong evidence of the importance of the study of neurodevelopmental genetic disorders for enhancing understanding of the complex manner in which initial genetic differences impact both behavior and processing strategies from infancy through adulthood. Our hope is that this issue will motivate further studies, informed by the genetic-developmental approach, on both Williams syndrome and other neurodevelopmental genetic disorders. PMID- 12730016 TI - Atypical mechanisms regulate the PMA-induced expression of IFN-gamma in a porcine trophectoderm cell line. AB - Interferon-gamma (IFN-gamma) is a major effector cytokine of the immune system with an expression pattern strictly restricted to cells of the lymphoid lineage. Several years ago, we reported that, during early pregnancy, the trophectoderm of the pig blastocyst, which represents a monolayer of polarized epithelial cells secretes high amount of IFN-gamma in a transient and developmentally regulated manner. In an effort to study the molecular basis of this atypical IFN-gamma gene expression, a pig trophectoderm cell line, TBA B4-3, was established in our laboratory. These cells developed a polarized phenotype with high transepithelial electrical resistance (TER) when grown on a microporous membrane. We found that treatment of polarized TBA B4-3 cells with the strong PKC agonist PMA induced, 3 4 days later, a transient IFN-gamma mRNA expression and vectorial IFN-gamma protein secretion. In order to better understand IFN-gamma gene regulation in TBA B4-3 cells, we examined in this system the effect of several drugs and factors known to affect the inducibility of this cytokine in T lymphocytes, the main source of IFN-gamma in the immunocompetent animal. We found that cyclosporine A (CsA) treatment of TBA B4-3 cells induces a partial inhibition of IFN-gamma secretion, thus indicating a minor role for the calcineurin signaling pathway in IFN-gamma expression. In addition, we found that although PMA alone can induce IFN-gamma secretion, the calcium ionophore A23187 synergizes with PMA for induction. We also analyzed by Southern blot the methylation status of a CpG dinucleotide in the 5' flanking region of IFN-gamma promoter and found that it was unmethylated in TBA B4-3 cells and in several pig epithelial cell lines that do not express IFN-gamma thus indicating the absence of correlation between demethylation and the ability to express IFN-gamma. Taken together, these results indicate that the mechanisms involved in IFN-gamma induction in TBA B4-3 cells are atypical compared to those presently known to operate in the T cell lineage. PMID- 12730018 TI - The roles of verbal short-term memory and working memory in the acquisition of grammar by children with Williams syndrome. AB - The roles of verbal short-term and working memory were examined in a sample of children with Williams syndrome (mean chronological age 10 years, 2 months) and a sample of grammar-matched children who are developing normally. Forward digit span, nonword repetition, and backward span were all found to be correlated with receptive grammatical ability in the sample of children with Williams syndrome, but not in the sample of children who are developing normally. The relation between working memory, as measured by backward digit span, and grammatical ability was found to be significantly stronger in children with Williams syndrome than in the control group. This finding highlights the possibility that children with Williams syndrome may rely on their working memory to a greater extent than children who are developing normally to learn grammar. Hierarchical regression analyses indicated receptive vocabulary may mediate the relations among forward digit span, backward digit span, and grammatical ability for the children with Williams syndrome. Phonological short-term memory, however, contributed independently to grammatical ability even after receptive vocabulary was taken into account. PMID- 12730019 TI - Early linguistic abilities of Italian children with Williams syndrome. AB - Previous studies of linguistic and memory abilities in Italian-speaking children with Williams syndrome (WS) and Down syndrome (DS)are briefly reviewed. New data on linguistic performance of 6 Italian children with WS between 3 and 6 years of age are presented and compared with data on linguistic performance of 6 children with DS selected from a larger sample and matched for chronological age and vocabulary size and of 6 typically developing (TD) younger children matched for mental age and vocabulary size. The language measures also included a parent report of early phrase structure, a naming test, and a sentence repetition task. Analyses revealed that the 3 groups of children were at the same productive vocabulary level, but showed different patterns in sentence production and repetition. Children with WS produced more complete sentences, similar to TD children at the same vocabulary size, whereas children with DS produced more telegraphic and incomplete sentences. The difference between children with DS and those with WS was more marked on the repetition task, suggesting that phonological short-term memory may play a greater role when sentence production is measured through repetition. In addition, qualitative analysis of errors produced in the repetition test revealed interesting differences among the 3 groups. These results from younger children confirm and extend previous findings with older children and adolescents with WS. They further suggest that the apparently spared linguistic abilities of children with WS could emerge as an artifact of comparisons made to children with DS, whose sentence production competence is more compromised relative to other verbal and nonverbal abilities. PMID- 12730020 TI - Morphological abilities of Hebrew-speaking adolescents with Williams syndrome. AB - In recent years research has focused on the exact nature of the linguistic skills that individuals with Williams syndrome (WS) exhibit. This work has resulted in controversial positions, with an increasing number of studies casting doubt on previous claims of superior linguistic competence for individuals with WS. This study investigated morphosyntactic knowledge in Hebrew-speaking adolescents with WS. The participants 'performance was compared to 2 groups of typically developing mental age-matched controls. Participants and controls were tested on experimental tasks designed to investigate knowledge of morphology. The findings suggest that individuals with WS have good control over the basic consonantal root structure of Hebrew words. However, rather poor performance was evident on other morphological paradigms. We conclude that there is little evidence from Hebrew to support a selective preservation of grammatical competence in individuals with WS. PMID- 12730021 TI - Can adolescents with Williams syndrome tell the difference between lies and jokes? AB - A group of adolescents with Williams syndrome (WS) was compared to matched groups of adolescents with Prader-Willi syndrome and nonspecific mental retardation on a task that tested the ability to distinguish different forms of nonliteral language. Participants listened to stories that ended in either a lie or an ironic joke. They were asked to decide the form of the nonliteral utterance and justify their responses. Almost none of the participants in any of the groups were able to correctly classify the ironic jokes, instead judging them to be lies because they did not correspond to reality. Their errors were similar to those made by younger normally developing children, but contrasted with those made by brain-damaged adults. These data are taken as further evidence that neurodevelopmental disorders are quite different from acquired brain disorders and require different neuropsychological models. The findings from this study also have important implications for considering the difficulties that adolescents with WS and other disorders will have in everyday social situations, especially among peers. PMID- 12730022 TI - Objects, motions, and paths: spatial language in children with Williams syndrome. AB - The acquisition of spatial language is often assumed to be built upon an early- emerging system of nonlinguistic spatial knowledge. We tested this relationship by examining spatial language in children with Williams syndrome (WS), a rare genetic disorder that gives rise to severe nonlinguistic spatial de deficits together with relatively spared language. Twelve children with WS, 12 normally developing mental-age matched children, and 12 normal adults described 80 videotaped motion events. Children with WS showed substantial control over key linguistic components of the motion event, including appropriate semantic and syntactic encoding of Figure and Ground objects, Manner of Motion, and Path. The expression of Path, although surprisingly spared, was more fragile among children with WS in contexts plausibly related to their nonlinguistic spatial deficit. The results show strong preservation of the formal aspects of spatial linguistic knowledge and suggest that the nonlinguistic spatial deficits shown by children with WS have, at most, limited effects on their spatial language. These findings have implications for the relationship between spatial language and other aspects of spatial cognition. PMID- 12730023 TI - Neurobiological models of visuospatial cognition in children with Williams syndrome: measures of dorsal-stream and frontal function. AB - We examine hypotheses for the neural basis of the profile of visual cognition in young children with Williams syndrome (WS). These are:(a)that it is a consequence of anomalies in sensory visual processing,(b)that it is a de.cit of the dorsal relative to the ventral cortical stream,(c)that it reflects de.cit of frontal function, in particular of frontoparietal interaction, and (d)that it is related to impaired function in the right hemisphere relative to the left. The tests reported here are particularly relevant to hypotheses 2 and 3. They form part of a more extensive program of investigating visual, visuospatial, and cognitive function in large group of children with WS children, aged 8 months to 15 years. To compare performance across tests, avoiding floor and ceiling effects, we have measured performance in children with WS in terms of the "age equivalence " for typically developing children. In this article the relation between dorsal and ventral function is tested by motion and form coherence thresholds, respectively. We confirm the presence of a subgroup of children with WS who perform particularly poorly on the motion (dorsal) task. However, such performance is also characteristic of normally developing children up to 5 years; thus the WS performance may reflect an overall persisting immaturity of visuospatial processing that is particularly evident in the dorsal stream. Looking at the performance on the global coherence tasks of the entire WS group, we find that there is also a subgroup who have both high form and motion coherence thresholds, relative to the performance of children of the same chronological age and verbal age on the British Picture Vocabulary Scale, suggesting a more general global processing deficit. Frontal function was tested by a counterpointing task, ability to retrieve a ball from a "detour box," and the Stroop--like "day.night " task, all of which require inhibition of a familiar response. When considered in relation to overall development as indexed by vocabulary, the day.night task shows little specific impairment, the detour box shows a significant delay relative to controls,and the counterpointing task shows a marked and persistent deficit in many children. We conclude that frontal control processes show most impairment in WS when they are associated with spatially directed responses, reflecting a deficit of frontoparietal processing. However, children with WS may successfully reduce the effect of this impairment by verbally mediated strategies. On all these tasks we find a range of difficulties across individual children and a small subset of children with WS who show very good performance, equivalent to chronological age norms of typically developing children. Overall, we conclude that children with WS have specific processing difficulties with tasks involving frontoparietal circuits within the spatial domain. However, some children with WS can achieve similar performance to typically developing children on some tasks involving the dorsal stream although the strategies and processing may be different in the 2 groups. PMID- 12730024 TI - Visuospatial cognition in Williams syndrome: reviewing and accounting for the strengths and weaknesses in performance. AB - Individuals with Williams syndrome typically show relatively poor visuospatial abilities in comparison to stronger verbal skills. However, individuals' level of performance is not consistent across all visuospatial tasks. The studies assessing visuospatial functioning in Williams syndrome are critically reviewed, to provide a clear pattern of the relative difficulty of these tasks. This prompts a possible explanation of the variability in performance seen, which focuses on the processing demands of some of these tasks. Individuals with Williams syndrome show an atypical processing style on tests of construction, which does not affect tests of perception. PMID- 12730025 TI - Implicit learning in children and adults with Williams syndrome. AB - In comparison to explicit learning, implicit learning is hypothesized to be a phylogenetically older form of learning that is important in early developmental processes (e.g., natural language acquisition, socialization)and relatively impervious to individual differences in age and IQ. We examined implicit learning in a group of children and adults (9.49 years of age)with Williams syndrome (WS)and in a comparison group of typically developing individuals matched for chronological age. Participants were tested in an artificial-grammar learning paradigm and in a rotor-pursuit task. For both groups, implicit learning was largely independent of age. Both groups showed evidence of implicit learning but the comparison group outperformed the WS group on both tasks. Performance advantages for the comparison group were no longer significant when group differences in working memory or nonverbal intelligence were held constant. PMID- 12730026 TI - Dethroning the myth: cognitive dissociations and innate modularity in Williams syndrome. AB - Despite increasing empirical data to the contrary, it continues to be claimed that mor-phosyntax and face processing skills of people with Williams syndrome are intact. This purported intactness, which coexists with mental retardation, is used to bolster claims about innately specified, independently functioning modules, as if the atypically developing brain were simply a normal brain with parts intact and parts impaired. Yet this is highly unlikely, given the dynamics of brain development and the fact that in a genetic microdeletion syndrome the brain is developing differently from the moment of conception, throughout embryogenesis, and during postnatal brain growth. In this article, we challenge the intactness assumptions, using evidence from a wide variety of studies of toddlers, children, and adults with Williams syndrome. PMID- 12730027 TI - Attentional characteristics of infants and toddlers with Williams syndrome during triadic interactions. AB - Two studies were conducted to consider the looking behavior of infants and toddlers with Williams syndrome (WS). In Study 1,the looking behavior of a 10 month-old girl with WS during play sessions with her mother and with a stranger was compared to that of 2 groups of infants who were developing normally (ND),1 matched for chronological age and the other for developmental age. The infant with WS spent more than twice as much time looking at her mother as the infants in either contrast group did. She also spent twice as much time looking at the stranger. In addition, during 78%of this time, her gaze at the stranger was coded as extremely intense. Looks of this intensity were virtually never made by the ND infants. In Study 2,the looking behavior of 31 individuals with WS ages 8 to 43 months during a genetics evaluation was compared to that of 319 control children in the same age range (242 with developmental delay due to causes other than WS).Twenty-three of the 25 participants with WS aged 33 months or younger demonstrated extended and intense looking at the geneticist. In contrast, none of the control participants looked extensively or intently at the geneticist. Findings are discussed in the context of previous research on arousal and focused attention during normal development and on temperament and personality of older children and adults with WS. It is argued that the unusual looking patterns evidenced by infants and toddlers with WS presage the unusual temperament and personality of older individuals with WS, and the possibility of a genetic basis for these behaviors is addressed. PMID- 12730028 TI - Distinctive personality characteristics of 8-, 9-, and 10-year-olds with Williams syndrome. AB - Although previous research and clinical observation have indicated that individuals with Williams syndrome have a distinctive personality, an empirically derived personality profile has not been developed. The objective of the current investigation was to develop a personality profile that is descriptive of and distinctive to children with Williams syndrome. Participants were 23 8- to 10 year-old children with Williams syndrome and 20 8- to 10-year-old children with developmental disabilities of other etiologies. Participant groups had equivalent intellectual abilities. Parents completed measures of childhood temperament (Children's Behavior Questionnaire [CBQ]) and personality (parent report, short form of Multidimensional Personality Questionnaire [MPQ]). Using group comparisons and signal detection theory, we contrasted the personality characteristics of children with Williams syndrome and children with developmental disabilities of other etiologies. On the CBQ, high mean ratings on shyness (reverse-coded) and empathy together characterized 96% of the children in the Williams syndrome group, but only 15% of the mixed etiology group. On the MPQ, high ratings on items measuring certain characteristics combined (gregarious, people-oriented, tense, sensitive, and visible) were characteristic of 96% of the Williams syndrome group but only 15% of the mixed etiology group. The personality profiles emerging from the CBQ and MPQ provide a crucial step toward investigations of genotype/phenotype relations. PMID- 12730029 TI - Anxiety, fears, and phobias in persons with Williams syndrome. AB - Although much research has focused on the cognitive-linguistic profile associated with Williams syndrome, studies have yet to follow up on preliminary observations suggesting increased anxiety and fears in persons with this disorder. To this aim, Study 1 compared fears in 120 participants with Williams syndrome to 70 appropriately matched persons with mental retardation of mixed etiologies. Study 2 assessed differences in parent versus child reports of fears in 36 Williams syndrome and 24 comparison group parent-child dyads. In Study 3, rates of phobia and other anxiety disorders were assessed in standardized psychiatric interviews with the parents of 51 individuals with Williams syndrome. Relative to their counterparts, persons with Williams syndrome had significantly more fears as well as a wider range of frequently occurring fears, as reported by either parents or participants themselves. Children in both groups reported more fears than their parents. Whereas generalized and anticipatory anxiety were found in 51% to 60% of the sample with Williams syndrome, specific phobia was more prevalent, with 96% showing persistent and marked fears and 84% avoiding their fears or enduring them with distress. The feasibility of cognitive-behavioral treatments for phobia is discussed, as are implications for future research. PMID- 12730030 TI - Glucose and lipid metabolism in young lean normotensive males with the G protein beta3 825T-allele. AB - The 825T-allele of the C825T polymorphism in GNB3, the gene for the G protein beta3 subunit, has been reported to be associated with essential hypertension and obesity. Expression of Gbeta3s, the gene product of GNB3 associated with the GNB3 825T-allele, causes increased signal transduction which may contribute to pathogenetic mechanisms ultimately resulting in hypertension and obesity. Given the known involvement of heterotrimeric G proteins in insulin secretion and insulin action on the cellular level, we analysed insulin sensitivity in each 15 young lean normotensive males with TC- and CC-genotypes, respectively. Blood glucose and serum insulin samples were taken during a standard oral glucose tolerance test. Insulin-stimulated glucose disposal was analysed by euglycemic hyperinsulinemic clamp. Both groups did not differ with regard to the time courses for glucose or insulin concentrations in the oral glucose tolerance test. Furthermore, insulin-stimulated glucose disposal was virtually independent of genotype. The TC-genotype is not associated with a primary defect in insulin secretion or sensitivity suggesting that obesity and hypertension in carriers of 825T do not likely result from primary alterations in glucose and insulin homeostasis. However, GNB3 825T-associated obesity may predispose to insulin resistance, an issue which remains to be investigated. Furthermore, fasting cholesterol was significantly higher in TC compared to CC genotype (4.71 versus 3.96 mmol/l; p = 0.007) suggesting that enhanced G protein signalling might be associated with alterations of cholesterol metabolism. PMID- 12730031 TI - Temporomandibular joint function in patients with longstanding rheumatoid arthritis - I. Role of periodontal status and prosthetic care - a clinical study. AB - PURPOSE: Temporomandibular joint (TMJ) dysfunction in rheumatoid arthritis (RA) occurs in 2 % to 86 % of RA patients. Dental factors possibly contributing to the development of TMJ dysfunction in RA patients have rarely been investigated in controlled studies. The present clinical study aimed 1) to compare patients with active, longstanding RA and healthy control subjects matched for age, sex, periodontal risk factors, dental and prosthetic status in order to obtain data on the prevalence of TMJ dysfunction in dentate RA patients and 2) to investigate a possible relationship between RA activity, general functional state and the severity of TMJ involvement. METHODS: 50 RA patients (38 F, 12 M; 54 +/- 9 years) were compared with 101 control subjects (76 F, 25 M; 54 +/- 11 years) with regard to dental, periodontal and prosthetic status and clinical TMJ findings as measured by the Helkimo indices. Clinical evaluation of RA patients included serological parameters, pain as measured by visual analog scale (VAS), a 28-joint count, a radiological destruction score, a functional status and measurement of grip strength. RESULTS: The sum of carious, missing and filled teeth was similar in both groups. RA patients had more missing teeth (p < 0.01), more gingival bleeding, deeper pockets and more attachment loss (p < 0.0001). They showed no differences with regard to the mean number of occluding pairs of teeth, tooth support, the percentage of dentures, the grade of prosthetic support. 36 % of RA patients had a unilaterally shortened dental arch compared with 11.9 % in controls (p < 0.05). 32 % of RA patients and 27.7 % of the control subjects reported TMJ or facial pain. The mean VAS was 50 +/- 19 for RA patients and 52 +/ 21 for controls. The anamnestic data and the clinical symptoms grouped according to the Helkimo index showed no significant differences between both subject groups. However, the maximal mouth opening capacity in RA patients was significantly lower (40.6 +/- 6.5 mm) than in controls (45.8 +/- 5.5 mm; p < 0.001). Analysis of the Helkimo symptom groups revealed a significantly reduced mobility index in the RA group and impaired TM-joint function in controls (p < 0.05). Grip strength was significantly correlated with mouth opening capacity, TMJ pain with tooth support. CONCLUSION: The prevalence of TMJ dysfunction in dentate patients with longstanding RA does not exceed that of healthy controls when structural risk factors predisposing to the development of temporomandibular dysfunction are taken into consideration. Maintaining adequate tooth support might help to prevent progressive TMJ impairment in the course of disease. PMID- 12730032 TI - Mistletoe in cancer - a systematic review on controlled clinical trials. AB - BACKGROUND: Mistletoe preparations are among the most widely used unconventional cancer therapies in Central Europe. Their clinical effectiveness, however, is controversial. OBJECTIVE: To investigate whether prospective controlled clinical trials provide evidence for efficacy of mistletoe therapy in cancer. DESIGN: Systematic review. MATERIAL AND METHODS: Search of 11 electronic databases, reference lists and expert consultations. Criteria based analysis was performed to assess methodological quality of the studies. RESULTS: Twenty-three studies were identified: 16 randomized, 2 quasi-randomized and 5 non-randomized. Cancer sites included breast, lung, stomach, colon, rectum, head and neck, kidney, bladder, melanoma, glioma, and genital. Among these studies, statistically significant positive outcomes were reported for survival (n = 8), tumor remission (n = 1), overall quality of life (QOL) (n = 3), and QOL in relation to side effects during cytoreductive therapy (n = 3). Further, positive trends were reported for survival (n = 8), disease-free-survival (n = 1), and tumor remission (n = 2). Several studies reported no effect on survival (n = 4), disease-free survival (n = 1), recurrence (n = 2), remission (n = 3), and QOL (n = 1). One study showed a negative trend for disease-free-survival. However, methodological quality of the studies was sometimes far below the standard that is today regarded as optimal or necessary. In view of substantial heterogeneity of the studies and potential positive and negative biases, we considered effect size estimation by quantitative synthesis to be unreliable and decided on a non quantitative synthesis and discussion. Mistletoe therapy was well tolerated, and no major side effects were noted. CONCLUSIONS: Among 23 identified studies evaluated for clinically relevant outcome measures, 12 studies showed one or more statistically significant, positive results, another 7 studies showed at least one positive trend, 3 showed no effect and 1 had a negative trend. All studies, however, suffered from methodological shortcomings to some degree, and many of the studies are not conclusive. As several reasonably well conducted studies indicate beneficial effects, further properly designed trials should be encouraged. Future controlled studies should take into account the methodological limitations and potential biases of these past mistletoe trials. PMID- 12730033 TI - Heat-shock protein HSP70-2 genotypes in patients with Crohn's disease: a more severe clinical course with intestinal complications in presence of the PstI polymorphism. AB - BACKGROUND: 70 kDa Heat shock proteins are involved in mucosal protecting reactions in the gut of patients with inflammatory bowel disease. Recently, a single nucleotide polymorphism (PstI, nucleotide 1267) was associated with intestinal perforations and formation of abscesses and fistulas in Japanese patients with Crohn's disease. Our purpose was to evaluate this phenomenon in Caucasian patients with Crohn's disease and to verify the clinical importance of this polymorphism. PATIENTS AND METHODS: 61 consecutive patients with Crohn's disease and 61 healthy control persons were examined. After DNA extraction and PCR amplification spanning the PstI-site, restriction fragment length polymorphism analyses (RFLP) were performed. Homozygous and heterozygous genotypes (AA, AB, BB) were then correlated with the clinical characteristics of the patients, especially with their intestinal complications. RESULTS: Intestinal perforations and formation of fistulas, abscesses and conglomerate tumors were significantly associated with allele B (p = 0.04). Patients with genotype BB showed the highest prevalence for surgical interventions (82%), whereas patients with genotype AA had the lowest prevalence (56%). Onset of disease, the need for immunosuppressive therapy and the occurrence of extraintestinal manifestations did not differ between the three genotypes. Allele A was significantly associated with combined involvement of ileum and colon. CONCLUSION: Caucasian patients carrying the HSP70-2 PstI-polymorphism seem to have a more severe form of Crohn's disease (perforations, abscesses, fistulas, conglomerate tumors). Especially, the homozygous genotype (BB) predisposes for a clinical course with high risk of surgical intervention. PMID- 12730034 TI - Re-operation for complicated secondary peritonitis - how to identify patients at risk for persistent sepsis. AB - INTRODUCTION: There is an ongoing dispute on the benefit of planned relaparotomy for patients with diffuse peritonitis. SETTING: Surgery Department, university hospital. PATIENTS: 145 patients with diffuse peritonitis treated with planned relaparotomy were analysed for APACHE II, MOF- and MODS-score (Goris and Marshall), complications, outcome and clinical/laboratory factors indicating intra-abdominal compartment syndrome (positive endexpiratory pressure (PEEP), central venous pressure (CVP), creatinine, blood urea nitrogen (BUN)) after termination of planned relaparotomy. Statistical analysis of data (mean and standard deviation) was performed using Mann-Whitney, chi-square, ANOVA and multiple regression analysis. RESULTS: The overall mortality was 29.7% and APACHE II score on admission 16.7 +/- 8.3. In 107 patients (mortality 17.8%) closure of the abdomen was achieved at termination of planned relaparotomy, 20 patients (mortality 30%) were treated with mesh closure and in 18 patients (mortality 100%) closure of the abdomen was not feasible. After closure of the abdomen 39 patients showed signs of persistent sepsis. Patients who were explored had a mortality of 37.5% and without re-exploration a mortality of 67%. BUN, PEEP and CVP were significantly different in survivors and non-survivors. Independent predictors of outcome were closure of the abdomen, complications, APACHE II and MOF scores. CONCLUSION: Patients with planned relaparotomy for diffuse peritonitis are not a uniform group and differ in mortality depending on source control and closure of the abdomen. Patients with persistent sepsis after termination of planned relaparotomy may be recognized by clinical and laboratory parameters and benefit from a timely reexploration. The decision when to close the abdomen may not only be based on intraperitoneal findings but also on the existence and level of organ failure. PMID- 12730035 TI - Intestinal anisakiasis mimicking intestinal obstruction. AB - We report one case of acute abdomen after the ingestion of raw or undercooked fish containing nematode larvae of the genus Anisakis. Early diagnosis is very important as it could prevent unnecessary surgical procedures since the symptoms of intestinal anisakiasis may mimic other illnesses such as appendicitis, ileitis or peritonitis. We suggest that anisakiasis should be considered in the differential diagnosis of patients with acute abdomen. PMID- 12730036 TI - Accuracy and precision of the PEAK Performance Technologies Motion Measurement System. AB - This study evaluated the accuracy and precision of the PEAK Performance Technologies, lnc.'s motion analysis system for three-dimensional angle reconstruction. Pendular motion of a bar, on which 18 retroreflective markers were mounted, was videotaped at three different orientations (parallel, and rotated 30 degrees right and left) to a plane at which two standard video cameras were aimed. The videotaped motion was digitized off-line, and 32 angles between the 18 markers were calculated. intraclass correlation coefficients (ICCs) were calculated between trials within each pendulum orientation and across orientations to determine system precision, and between randomly selected trials and actual angles to determine accuracy. lCCs were in all cases greater than.99. Within-trial standard deviations ranged between 0.05-0.8 degrees for the different angles. Deviations from the actual angle averaged 0.0-0.8 degrees across all angles and orientations. The results indicate that accurate and reliable angular measurements can be made with this motion analysis system. PMID- 12730038 TI - Asymmetries in the regulation of visually guided aiming. AB - An experiment was conducted to examine the contribution of sensory information to asymmetries in manual aiming. Movements were performed in four vision conditions. In the full-vision condition (FV), subjects were afforded vision of both the hand and the target throughout the course of the movement. In the ambient-illumination off condition (AO), the room lights were extinguished at movement initiation, preventing vision of the moving limb. In the target-off (TO) condition, the target was extinguished upon initiation of the movement. In a no-vision (NV) condition, ambient illumination was removed and the target was extinguished upon initiation of the response movement. Results indicated that accuracy was superior in the full-vision and target-off conditions and when movements were made by the right hand. Movements made by the right hand were also of shorter mean duration. The magnitudes of performance asymmetries were uninfluenced by vision condition. Analyses of movement kinematics revealed that movements made in conditions in which there was vision of the limb exhibited a greater number of discrete modifications of the movement trajectory. On an individual-trial basis, no relationship existed between accuracy and the occurrence of discrete modifications. These data suggest that although vision greatly enhances accuracy, discrete modifications subserved by vision reflect the imposition of nonfunctional zero-order control processes upon continuous higher-order control regimes. PMID- 12730037 TI - Space-time accuracy of rapid movements. AB - The space-time accuracy of an elbow flexion movement task was examined in two experiments over a range of motion extents (1 degrees through 100 degrees ) and short-duration movement times (100, 125, 150, and 400 ms). Nonlinear speed accuracy functions emerged for both spatial and temporal error over all the movement conditions examined. The results showed that the timing error and spatial error had a high degree of complementarity as predicted by a space-time model of the speed-accuracy relation (Hancock & Newell, 1985). The findings confirm that the frame of reference for measuring movement error determines in part the error functions observed. PMID- 12730039 TI - Impulse variability in isometric tasks. AB - An isometric elbow flexion task was used in two experiments that examined the influence of force-production characteristics on impulse variability. Impulse size was held constant while peak force, time to peak force, rate of force, and, hence, the shape of the criterion force-time curve were manipulated. The results indicated that changes in the force-time curve under conditions of equal impulse bring about systematic changes in impulse variability, and this effect is more pronounced for larger impulse conditions. The inability of existing functions to account for the peak force variability findings led to the generation of a new predicted force variability function. The proposed function accounts for changes in the standard deviation and coefficient of variation of peak force, impulse, and rate of force over a range of force-time conditions. PMID- 12730040 TI - Same-hand and different-hand finger pairings in two-choice reaction time: presence or absence of response competition? AB - Several two-choice reaction time experiments have compared conditions in which the two possible responses were from the same hand (same-hand pairing) or from different hands (different-hand pairing). Studies that used only the two relevant fingers on response keys reported shorter reaction times for the different-hand pairing. In other studies, two additional but irrelevant fingers were also in contact with response keys. These fingers were irrelevant in the sense that they never were required to respond. With this procedure, equivalent reaction times were found between same-hand and different-hand pairings. Reeve and Proctor (1988) recently have argued that using only two fingers results in response competition between the two fingers from the same hand, yielding shorter reaction times for the different-hand pairing condition. In contrast, when four fingers are placed on response keys, response competition should be absent for both the same-hand and the different-hand pairing conditions, resulting in equivalent reaction times. In the present work, reaction times associated with the same-hand pairing condition remained unchanged, irrespective of the number of fingers positioned on keys. In the different-hand pairing condition, reaction times were found to be longer when four fingers were used than when only two fingers rested on response keys. Thus, when four fingers are placed on keys, response competition appears to be present rather than absent. Other results showed that the response competition found in the different-hand pairing condition decreases with practice. PMID- 12730041 TI - Intermittency in human manual tracking tasks. AB - We confirm Craik's (1947) observation that the human manual1y tracking a visual target behaves like an intermittent servo-control1er. Such tracking responses are indicative of "sampled" negative-feedback control but could be the result of other, continuous, mechanisms. Tracking performance therefore was recorded in a task in which visual feedback of the position of the hand-held joystick could be eliminated. Depriving the subjects of visual feedback led to smoother tracking and greatly reduced the signal power of their responses between 0.5-1.8 Hz. Their responses remained intermittent when they used feedback of their own position but not of the target to track a remembered (virtual) target. Hence, intermittency in tracking behavior is not exclusively a signature of visual feedback control but also may be a sign of feedback to memorized waveforms. Craik's (1947) suggestion that the intermittency is due to a refractory period following each movement was also tested. The errors measured at the start of each intermittent response, during tracking of slow waveforms, showed evidence of a small error deadzone (measuring 0.7 cm on the VDU screen or 0.80 degrees at the eye). At higher target speeds, however, the mean size of starting errors increased, and the upper boundary of the distribution of starting error was close to that expected of a refractory delay of approximately 170 ms between responses. We consider a model of the control system that can fit these results by incorporating an error deadzone within a feedback control loop. We therefore propose that the initiation of intermittent tracking responses may be limited by a positional error deadzone and that evidence for a refractory period between successive corrective movements can be satisfied without evoking an explicit timing or sampling mechanism. PMID- 12730042 TI - Above- and belowground biomass and net primary production in a 73-year-old Scots pine forest. AB - We estimated above- and belowground biomass and net primary production (NPP) of a 73-year-old Scots pine (Pinus sylvestris L.) forest stand in the Belgian Campine region. Total biomass for the stand was 176 Mg ha(-1), of which 74.4% was found in stems. The root system contained 12.6% of total biomass, most of it in coarse roots (> 5 mm). Fine roots (< 5 mm) comprised only about 1.7% of total biomass, and more than 50% of fine root biomass was retrieved in the litter layer and the upper 15 cm of the mineral soil. The ratio of belowground biomass to aboveground biomass was 0.14, which is lower than that of other Scots pine forests and other coniferous forests. Between 1995 and 2001, mean annual NPP was 11.2 Mg ha(-1) year(-1), of which 68.7% was allocated to aboveground compartments. Stems, needles and cones made relatively high contributions to total NPP compared with branches. However, branch NPP was possibly underestimated because litterfall of big branches was neglected. The proportion of total NPP in belowground components was 31.3%. Coarse root NPP (2% of total) was low compared with its biomass. Fine root NPP was 3.3 Mg ha(-1) year(-1), representing about 29.5% of total NPP; however, the estimate of fine root NPP is much more uncertain than NPP of aboveground compartments. The ratio NPP/GPP (gross primary production) was 0.32, which was low compared with other coniferous forests. PMID- 12730043 TI - Differences in leaf phenology between juvenile and adult trees in a temperate deciduous forest. AB - In a deciduous forest, differences in leaf phenology between juvenile and adult trees could result in juvenile trees avoiding canopy shade for part of the growing season. By expanding leaves earlier or initiating senescence later than canopy trees, juvenile trees would have some period in high light and therefore greater potential carbon gain. We observed leaf phenology of 376 individuals of 13 canopy tree species weekly over 3 years in a deciduous forest in east central Illinois, USA. Our objectives were: (1) to quantify for each species the extent of differences in leaf phenology between juvenile and conspecific adult trees; and (2) to determine the extent of phenological differences between juvenile Aesculus glabra Willd. and Acer saccharum Marsh. trees in understory and gap microhabitats. All species displayed phenological differences between life stages. For 10 species, bud break was significantly earlier, by an average of 8 days, for subcanopy individuals than for canopy individuals. In 11 species, completion of leaf expansion was earlier, by an average of 6 days, for subcanopy individuals than for canopy individuals. In contrast, there were no significant differences between life stages for start of senescence in 10 species and completion of leaf drop in nine species. For eight species, leaf longevity was significantly greater for subcanopy individuals than for canopy individuals by an average of 7 days (range = 4-10 days). Leaf phenology of subcanopy individuals of both Aesculus glabra and Acer saccharum responded to gap conditions. Leaf longevity was 11 days less in the understory than in gaps for Aesculus glabra, but 14 days more in the understory than in gaps for Acer saccharum. Therefore, leaf phenology differed broadly both between life stages and within the juvenile life stage in this community. A vertical gradient in temperature sums is the proposed mechanism explaining the patterns. Temperature sums accumulated more rapidly in the sheltered understory than in an open elevated area, similar to the canopy. Early leaf expansion by juvenile trees may result in a period of disproportionately higher carbon gain, similar to gains made during summer months from use of sun flecks. PMID- 12730044 TI - Ultraviolet-B radiation alters phenolic salicylate and flavonoid composition of Populus trichocarpa leaves. AB - We investigated foliar phenolic composition of field- and greenhouse-grown Populus trichocarpa Torr. & A. Gray (black cottonwood) ramets subjected to near zero (0x), ambient (1x) or twice ambient (2x) concentrations of biologically effective ultraviolet-B (UV-B) radiation. After a 3-month treatment period, several age classes of foliage samples were harvested and the phenolic compounds extracted, separated by high performance liquid chromatography and identified and quantified by diode-array spectrometry and mass spectrometry. Foliar phenolic concentration was greater in 1x- and 2x-treated tissue than in 0x-treated tissue. Phenolic compounds that increased in response to UV-B radiation were predominantly flavonoids, primarily quercetin and kaempferol glycosides. Enhancement of UV-B radiation from 1x to 2x ambient concentration did not result in further flavonoid accumulation in either greenhouse or field ramets; however, a non-flavonoid phenolic glycoside, salicortin, increased in response to an increase in UV-B radiation from 1x to 2x ambient concentration. Increased salicortin concentrations accounted for at least 30-40% of the total (5%) increase in UV-absorption potential of 2x-treated tissue. Because salicortin and other salicylates are important in plant-herbivore-predator relationships, these increases are discussed in the context of collateral feeding studies. We conclude that enhanced solar UV-B radiation may significantly alter trophic structure in some ecosystems by stimulating specific phenolic compounds. PMID- 12730045 TI - Reflectance of Alaskan black spruce and white spruce foliage in relation to elevation and latitude. AB - Leaf reflectance at visible and near-infrared wavelengths (400-1000 nm) is related primarily to pigmentation, leaf structure and water content, and is an important tool for studying stress physiology and relationships between plants and their growth environment. We studied reflectance of two co-occurring Alaskan conifers, black spruce (Picea mariana (Mill.) BSP) and white spruce (Picea glauca (Moench) Voss), at elevations from 60 to 930 m a.s.l. along a latitudinal gradient from 61 degrees to 68 degrees N. Black spruce samples were collected from 24 sites and white spruce from 30 sites. Overall, reflectance spectra of the two species were similar, but from 400 to 700 nm, needle reflectance was consistently higher in black spruce than in white spruce (all P 187 mmol N m-2). Leaf dry mass per area (DM area-1) and total chlorophyll concentration increased linearly with increasing leaf N, whereas chlorophyll a:b ratio declined. Net assimilation of CO2 (A(CO2)) and leaf water-use efficiency (WUE) reached maximum values in leaves with approximately 187 mmol N m-2. Nitrogen-deficient leaves exhibited small chloroplasts with no starch granules; grana and stroma lamellae that coincided with the accretion of numerous large plastoglobuli in the stroma disintegrated. High-N leaves had large chloroplasts with well-developed grana, stroma lamellae and starch granules that enlarged with increasing N concentration. The lack of an increase in A(CO2) capacity at leaf N concentrations above 187 mmol N m-2 appeared to be correlated with the presence of numerous large starch granules. PMID- 12730049 TI - Response of photosynthesis in second-generation Pinus radiata trees to long-term exposure to elevated carbon dioxide partial pressure. AB - Second-generation Pinus radiata D. Don trees, propagated from cuttings of 4-year old trees previously grown at ambient (36 Pa) and elevated (65 Pa) CO2 partial pressure (Ca) were grown under the same conditions in open-top chambers for a further year. As cuttings of the original trees, these second-generation trees were physiologically the same age as the first-generation trees with the only difference between the two being size. This allowed us to test the effects of tree size independently of age or duration of exposure. Total non-structural carbohydrate concentration, area-based nitrogen concentration, leaf mass per unit area and chlorophyll concentration measured in three foliage age cohorts were unaffected by either age or Ca. There were no signs of photosynthetic down regulation in trees grown at elevated Ca. When measured at the growth Ca, photosynthetic rate in young needles during summer, autumn and spring was 34, 43 and 38% higher, respectively, in trees grown at elevated Ca than in trees grown at ambient Ca. In older needles, the corresponding photosythetic rate increases were 26, 47 and 49%. Water-use efficiency, determined by stable carbon isotope analysis, was 49% higher in foliage in the elevated Ca treatment than in foliage in the ambient Ca treatment. This increase was entirely due to photosynthetic enhancement, because stomatal conductance did not differ between treatments. We conclude that down-regulation of photosynthesis at elevated Ca is related to tree size rather than tree age or duration of exposure, and that enhanced photosynthetic rates can be maintained while sink strength is high enough to use the excess photosynthates.elevated CO2, needle age, photosynthetic down regulation, photosynthetic enhancement, sink strength, water-use efficiency. PMID- 12730051 TI - CO2-dependent components of sinus arrhythmia from the start of breath holding in humans. AB - A substantial portion of sinus arrhythmia in conscious humans appears to be caused by the CO2-dependent central respiratory rhythm. Under some circumstances, therefore, sinus arrhythmia might indicate the presence of the central respiratory rhythm. Humans can voluntarily modify their central respiratory rhythm (e.g., by pacing breathing or by delaying or advancing breaths), but it is not clear what happens to it from the start of breath holding. In this study, we show that sinus arrhythmia persists from the start of breath holds prolonged by preoxygenation. We also show that some of the frequency components of sinus arrhythmia start within each subject's eupneic frequency range and change when end-tidal Pco2 is lowered or raised, as we would expect if the central respiratory rhythm continues from the start of breath holding. We discuss whether sinus arrhythmia can indicate if the central respiratory rhythm continues from the start of breath holding. PMID- 12730050 TI - Tetrahydrobiopterin restores endothelial dysfunction induced by an oral glucose challenge in healthy subjects. AB - An oral glucose challenge causes transient impairment of endothelial function, probably because of increased oxidative stress. During oxidative stress, endothelial nitric oxide (NO) synthase (eNOS) becomes uncoupled because of decreased bioavailability of tetrahydrobiopterin (BH4), an essential cofactor of eNOS. Therefore, we examined whether an acute supplement of BH4 could restore endothelial dysfunction induced by an oral glucose challenge. Healthy subjects were examined in 53 experiments. Forearm blood flow was measured by venous occlusion plethysmography. Dose-response studies were obtained during intra arterial infusion of serotonin to elicit endothelium-dependent, NO-specific vasodilation and during sodium nitroprusside (SNP) infusion to elicit endothelium independent vasodilation. Subjects were examined before (fasting) and 1 and 2 h after an oral glucose challenge (75 g) with serotonin (n = 10) and SNP (n = 8). On different days (6R)-5,6,7,8-tetrahydro-l-biopterin dihydrochloride (6R-BH4; n = 10), the active cofactor of eNOS or its stereoisomer (6S)-5,6,7,8-tetrahydro-l biopterin sulfate (6S-BH4; n = 10), which is inactive as a cofactor, was added 10 min (500 microg/min) before and during the 1-h postchallenge serotonin dose response study. In vitro studies showed that 6R-BH4 and 6S-BH4 were equipotent antioxidants. Serotonin response was reduced by 24 +/- 7% (at the highest dose) at 1 h postchallenge compared with fasting (P = 0.001) and was restored 2 h postchallenge. The reduction was reversed by the administration of 6R-BH4 but not by 6S-BH4. SNP responses were slightly increased 1 and 2 h postchallenge (increased by 15 +/- 13% at third dose 2 h postchallenge, P = 0.0001). An oral glucose challenge causes transient, NO-specific, endothelial dysfunction, which may be reversed by BH4. Transient postprandial endothelial dysfunction may be partly explained by reduced bioavailability of BH4 and NO. PMID- 12730052 TI - Positive inotropic effect of ceramide in adult ventricular myocytes: mechanisms dissociated from its reduction in Ca2+ influx. AB - Ceramide, a sphingolipid metabolite produced by activation of sphingomyelinase, has been previously shown to reduce L-type Ca2+ channel current (ICa,L) in adult rat ventricular myocytes; however, its effect on contractile function is unknown. In this study, we investigated the effects of ceramide on excitation-contraction coupling in adult ventricular myocytes and on left ventricular (LV) function in isolated hearts. Surprisingly, in patch-clamped myocytes, ceramide increased contraction concomitant with reductions in ICa,L. In intact myocytes, ceramide increased cell shortening (CS) concurrently with enhancing maximum rates of shortening and relaxation and the duration of contraction. Ceramide also increased the amplitudes of postrest potentiated (PRP) contraction. In fura-PE3 loaded myocytes, ceramide increased systolic Ca2+ and the magnitude and maximum rates of the rising and declining phases of Ca2+ transients. Ceramide-elicited decreases in magnitudes of PRP relative to steady-state contraction and the Ca2+ transient suggest an increased fractional Ca2+ release from the sarcoplasmic reticulum (SR). However, ceramide slightly reduced the caffeine-induced Ca2+ transient and had no significant effect on the amplitude of the PRP-elicited Ca2+ transient. Additionally, the ceramide-induced upward shift in the relationship of contraction and the Ca2+ transient and increase in the Ca2+ responsiveness of CS suggest an increase in myofilament Ca2+ sensitivity. In isolated hearts, ceramide increased LV developed pressure and maximum rates of contraction and relaxation at balloon volumes of 30-50 microl. In summary, regardless of decreasing ICa,L, ceramide elicits distinct positive inotropic and lusitropic effects, resulting probably from enhanced SR Ca2+ release and uptake, and increased Ca2+ sensitivity of ventricular myocytes. PMID- 12730053 TI - Redox factor-1 contributes to the regulation of progression from G0/G1 to S by PDGF in vascular smooth muscle cells. AB - Redox factor-1 (Ref-1/APE), a multifunctional DNA base excision repair and redox regulation enzyme, plays an important role in oxidative signaling, transcription factor regulation, and cell cycle control. We hypothesized that Ref-1 plays a regulatory role in smooth muscle cell (SMC) proliferation induced by PDGF. Ref-1 antisense oligodeoxynucleotides (AODN), which diminished the level of Ref-1 protein in SMCs by approximately 50%, inhibited PDGF-BB (composed of the homodimer of B-polypeptide chain)-induced [3H]thymidine incorporation compared with control oligodeoxynucleotides. Ref-1 AODN inhibited PDGF-BB-induced S phase entry by approximately 63%, which was overcome by overexpression of Ref-1 by adenoviral-mediated gene transfer. Overexpression of Ref-1 alone without PDGF enhanced SMC entry into the S phase. Furthermore, decreasing Ref-1 protein by treatment of SMCs with Ref-1 AODN, or by immunodepletion of Ref-1 from nuclear extracts, inhibited PDGF-BB-induced activator protein-1 (AP-1) DNA binding activity. Chemical reduction restored the AP-1 DNA binding in Ref-1-depleted nuclear extracts. These results suggest that Ref-1 contributes to the regulation of PDGF-BB-stimulated cell cycle progression from G0/G1 to S in SMCs, with one of the possible steps being redox-regulation of AP-1 by Ref-1 protein. PMID- 12730054 TI - Evidence for ERK1/2 activation by thrombin that is independent of EGFR transactivation. AB - Thrombin is involved in abnormal proliferation of vascular smooth muscle cells (VSMCs) associated with pathogenic vascular remodeling. Thrombin stimulation results in extracellular signal-regulated kinase (ERK)1/2 activation through transactivation of the epidermal growth factor receptor (EGFR). Here, using specific antibodies and inhibitors, we investigated the thrombin-induced phosphorylation of Src family kinases, nonreceptor proline-rich tyrosine kinase (Pyk2), EGFR, and ERK1/2. Our results show that Src and Pyk2 are involved upstream of the EGFR transactivation that is required for ERK1/2 phosphorylation. The investigation of the role of intracellular calcium concentration ([Ca2+]i) and calcium mobilization with the Ca2+ chelator BAPTA and thapsigargin, respectively, indicated that thrombin- and thapsigargin-induced phosphorylation of the EGFR but not ERK1/2 is dependent on an increase in [Ca2+]i. Moreover, only after BAPTA-AM pretreatment was thrombin-induced activation of ERK1/2 partially preserved from the effects of EGFR and PKC inhibition but not Src family kinase inhibition. These results suggest that BAPTA, by preventing [Ca2+]i elevation, unmasks a new pathway of Src family kinase-dependent thrombin-stimulated ERK1/2 phosphorylation that is independent of EGFR and PKC activation. PMID- 12730055 TI - Novel 3D culture system for study of cardiac myocyte development. AB - Insufficient myocardial repair after pathological processes contributes to cardiovascular disease, which is a major health concern. Understanding the molecular mechanisms that regulate the proliferation and differentiation of cardiac myocytes will aid in designing therapies for myocardial repair. Models are needed to delineate these molecular mechanisms. Here we report the development of a model system that recapitulates many aspects of cardiac myocyte differentiation that occur during early cardiac development. A key component of this model is a novel three-dimensional tubular scaffold engineered from aligned type I collagen strands. In this model embryonic ventricular myocytes undergo a transition from a hyperplastic to a quiescent phenotype, display significant myofibrillogenesis, and form critical cell-cell connections. In addition, embryonic cardiac myocytes grown on the tubular substrate have an aligned phenotype that closely resembles in vivo neonatal ventricular myocytes. We propose that embryonic cardiac myocytes grown on the tube substrate develop into neonatal cardiac myocytes via normal in vivo mechanisms. This model will aid in the elucidation of the molecular mechanisms that regulate cardiac myocyte proliferation and differentiation, which will provide important insights into myocardial development. PMID- 12730056 TI - Pattern formation of vascular smooth muscle cells subject to nonuniform fluid shear stress: mediation by gradient of cell density. AB - Smooth muscle cells (SMCs) are organized in various patterns in blood vessels. Whereas straight blood vessels mainly contain circumferentially aligned SMCs, curved blood vessels are composed of axially aligned SMCs in regions with vortex blood flow. The vortex flow-dependent feature of SMC alignment suggests a role for nonuniform fluid shear stress in regulating the pattern formation of SMCs. Here, we demonstrate that, in experimental models with vascular polymer implants designed for the observation of neointima formation and SMC migration under defined fluid shear stress, nonuniform shear stress possibly plays a role in regulating the direction of SMC migration and alignment in the neointima of the vascular implant. It was found that fluid shear stress inhibited cell growth, and the presence of nonuniform shear stress influenced the distribution of total cell density and induced the formation of cell density gradients, which in turn directed SMC migration and alignment. In contrast, uniform fluid shear stress in a control model influenced neither the distribution of total cell density nor the direction of SMC migration and alignment. In both the uniform and nonuniform shear models, the gradient of total cell density was consistent with the alignment of SMCs. These observations suggest that nonuniform shear stress may regulate the pattern formation of SMCs, possibly via mediating the gradient of cell density in the neointima of vascular polymer implants. PMID- 12730057 TI - Activation of delta- and kappa-opioid receptors by opioid peptides protects cardiomyocytes via KATP channels. AB - To examine the receptor specificity and the mechanism of opioid peptide-induced protection, we examined freshly isolated adult rabbit cardiomyocytes subjected to simulated ischemia. Cell death as a function of time was assessed by trypan blue permeability. Dynorphin B (DynB) and Met5-enkephalin (ME) limitation of cell death (expressed as area under the curve) was sensitive to blockade by naltrindole (NTI, a delta-selective antagonist) and 5'-guanidinyl-17 (cyclopropylmethyl)-6,7-dehydro-4,5alpha-epoxy-3,14-dihydroxy-6,7-2',3' indolomorphinan (GNTI dihydrochloride, a kappa-selective antagonist): 85.7 +/- 2.7 and 142.9 +/- 2.7 with DynB and DynB + NTI, respectively (P < 0.001), 94.1 +/ 4.2 and 164.5 +/- 7.3 with DynB and DynB + GNTI, respectively (P < 0.001), 111.9 +/- 7.0 and 192.1 +/- 6.4 with ME and ME + NTI, respectively (P < 0.001), and 120.2 +/- 4.3 and 170.0 +/- 3.3 with ME and ME + GNTI, respectively (P < 0.001). Blockade of ATP-sensitive K+ channels eliminated DynB- and ME-induced protection: 189.6 +/- 5.4 and 139.0 +/- 5.4 for control and ME, respectively (P < 0.001), and 210 +/- 5.9 and 195 +/- 6.1 for 5-HD and ME + 5-HD, respectively (P < 0.001); 136.0 +/- 5.7 and 63.4 +/- 5.4 for control and ME, respectively (P < 0.001), and 144.6 +/- 4.5 and 114.6 +/- 7.7 for HMR-1098 and ME + HMR-1098, respectively (P < 0.01); 189.6 +/- 5.4 and 139.0 +/- 5.4 for control and ME, respectively (P < 0.001), and 210 +/- 5.9 and 195 +/- 6.1 for 5-HD and ME + 5-HD, respectively (P < 0.001); and 136.0 +/- 5.7 and 63.4 +/- 5.4 for control and ME, respectively (P < 0.001), and 144.6 +/- 4.5 and 114.6 +/- 7.7 for HMR-1098 and ME + HMR-1098, respectively (P < 0.01). We conclude that opioid peptide-induced cardioprotection is mediated by delta- and kappa-receptors and involves sarcolemmal and mitochondrial ATP-sensitive K+ channels. PMID- 12730058 TI - Interaction between nitric oxide and prostanoids in arterioles of rat cremaster muscle in vivo. AB - We studied in vivo interactions of nitric oxide (NO), oxidative stress, and prostanoids derived from the cyclooxygenase pathway in the arterioles studied by intravital microscopy in peripheral muscle. Topical administration of NO synthase (NOS) inhibitor Nomega-nitro-l-arginine (l-NNA) or cyclooxygenase inhibitor mefenamic acid (MA) alone leads to vasoconstriction. We found that l-NNA after MA induced an additional constriction, whereas MA after l-NNA induced a relative dilation. Therefore, an additional constriction was found when MA was administered after l-NNA in the presence of the thromboxane A2 synthase-PGH2 (TP) receptor antagonist SQ-29548. We also found a relative dilation when the TP receptor antagonist was administered after NOS inhibition by l-NNA. In the presence of superoxide dismutase and catalase, l-NNA-induced vasoconstriction is reduced, and the dilation observed after addition of MA in presence of the reactive oxygen species is no longer present. Taken together, these results showed that NO inhibition induced a shift in the synthesis or in the effects of cyclooxygenase products, in favor of constrictor prostanoids. This effect of NO inhibition disappears when reactive oxygen species are scavenged by superoxide dismutase and catalase. PMID- 12730059 TI - Role of hyaluronic acid glycosaminoglycans in shear-induced endothelium-derived nitric oxide release. AB - Endothelium-derived nitric oxide (NO) is synthesized in response to chemical and physical stimuli. Here, we investigated a possible role of the endothelial cell glycocalyx as a biomechanical sensor that triggers endothelial NO production by transmitting flow-related shear forces to the endothelial membrane. Isolated canine femoral arteries were perfused with a Krebs-Henseleit solution at a wide range of perfusion rates with and without pretreatment with hyaluronidase to degrade hyaluronic acid glycosaminoglycans within the glycocalyx layer. NO production rate was evaluated as the product of nitrite concentration in the perfusate and steady-state perfusion rate. The slope that correlates the linear relation between perfusion rate and NO production rate was taken as a measure for flow-induced NO production. Hyaluronidase treatment significantly decreased flow induced NO production to 19 +/- 9% of control (mean +/- SD; P < 0.0001 vs. control; n = 11), whereas it did not affect acetylcholine-induced NO production (88 +/- 17% of pretreatment level, P = not significant; n = 10). We conclude that hyaluronic acid glycosaminoglycans within the glycocalyx play a pivotal role in detecting and amplifying the shear force of flowing blood that triggers endothelium-derived NO production in isolated canine femoral arteries. PMID- 12730060 TI - Acrolein-induced vasomotor responses of rat aorta. AB - Acrolein is a highly reactive aldehyde pollutant and an endogenous product of lipid peroxidation. Increased generation of, or exposures to, acrolein incites pulmonary and vascular injury. The effects of acrolein on the vasomotor responses of rat aortic rings were studied to understand its mechanism of action. Incubation with acrolein (10-100 microM) alone did not affect the resting tone of aortic vessels; however, a dose-dependent relaxation of phenylephrine precontracted aortic rings was observed. Acrolein-induced relaxation was slow and time dependent and the extent of relaxation after 100 min of application was 44.7 +/- 4.1% (10 microM), 56.0 +/- 5.6% (20 microM), 61.0 +/- 7.9% (40 microM), and 96.1 +/- 2.1 (80 microM), respectively, versus 14.2 +/- 3.3% relaxation in the absence of acrolein. Acrolein-induced vasorelaxation was prevented by endothelial denudation and was abolished on pretreatment with the nitric oxide synthase inhibitor Nomega-nitro-L-arginine methyl ester, the guanylyl cyclase inhibitor 1H [1,2,4]oxidazolo[4,3-a]quinoxaline-1-one, or the cyclooxygenase inhibitor indomethacin. Inhibition of K+ channels (by tetrabutylammonium) or Na+-K+-ATPase (by ouabain) did not significantly prevent acrolein-mediated vasorelaxation. Exposure to acrolein in the presence or absence of other compounds elicited slow wave vasomotor effect in 77% of aortic vessels versus 1.4% in control. Vasomotor responses were also studied on aortic rings prepared from rats fed 2 mg. kg-1. day-1 acrolein for 3 alternate days by oral gavage. These vessels developed a significantly lower contractile response to phenylephrine compared with controls. Together, these results indicate that acute acrolein exposure evokes delayed vasorelaxation due to a nitric oxide- and prostacyclin-dependent mechanism, whereas in vivo acrolein exposure compromises vessel contractility. PMID- 12730061 TI - Extent of damage in ischemic, nonreperfused, and reperfused myocardium of anesthetized rats. AB - To investigate the localization of the earliest damage in ischemic and ischemic reperfused myocardium, anesthetized rats were subjected to coronary occlusion for 15, 30, 45, or 90 min. One-half of the animals in each group had no reperfusion, whereas the other half was reperfused for 14 min. With the use of histological methods, preferentially in the periphery of the area at risk, localized zones were detected that lacked the hypoxia-specific increase in NADH fluorescence. The extent of these areas displaying injured tissue was found to be significantly smaller in the ischemic-nonreperfused hearts than in the ischemic-reperfused organs (15-min ischemia: 0.22 +/- 0.12% vs. 43.0 +/- 5.0%; 30-min ischemia: 5.7 +/- 2.7% vs. 64.6 +/- 2.9%; 45-min ischemia: 5.6 +/- 1.2% vs. 66.0 +/- 7.5%; 90 min ischemia: 39.3 +/- 5.5% vs. 86.7 +/- 1.8% of the area at risk). The results point to a localized initiation of the damage close to the surrounding oxygen supplied tissue during ischemia and an expansion of this injury by intercellular actions into yet-intact areas upon reperfusion. PMID- 12730062 TI - Hyperhomocysteinemia leads to pathological ventricular hypertrophy in normotensive rats. AB - A recent report indicated that hyperhomocysteinemia (Hhe), in addition to its atherothrombotic effects, exacerbates the adverse cardiac remodeling seen in response to hypertension, a powerful stimulus for pathological ventricular hypertrophy. The present study was undertaken to determine whether Hhe has a direct effect on ventricular remodeling and function in the absence of other hypertrophic stimuli. Male Wistar-Kyoto rats were fed either an amino acid defined control diet or an intermediate Hhe-inducing diet. After 10 wk of dietary treatment, rats were subjected to echocardiographic assessment of left ventricular (LV) dimensions and systolic function. Subsequently, blood was collected for plasma homocysteine measurements, and the rats were killed for histomorphometric and biochemical assessment of cardiac remodeling and for in vitro cardiac function studies. Significant LV hypertrophy was detected by echocardiographic measurements, and in vitro results showed hypertrophy with significantly increased myocyte size in the LV and right ventricle (RV). LV and RV remodeling was characterized by a disproportionate increase in perivascular and interstitial collagen, coronary arteriolar wall thickening, and myocardial mast cell infiltration. In vitro study of LV function demonstrated abnormal diastolic function secondary to decreased compliance because the rate of relaxation did not differ between groups. LV systolic function did not vary between groups in vitro. In summary, in the absence of other hypertrophic stimuli short-term intermediate Hhe caused pathological hypertrophy and remodeling of both ventricles with diastolic dysfunction of the LV. These results demonstrate that Hhe has direct adverse effects on cardiac structure and function, which may represent a novel direct link between Hhe and cardiovascular morbidity and mortality, independent of other risk factors. PMID- 12730063 TI - Age-associated impairment in TNF-alpha cardioprotection from myocardial infarction. AB - Age-associated dysfunction in cardiac microvascular endothelial cells with impaired induction of cardioprotective platelet-derived growth factor (PDGF) dependent pathways suggests that alterations in critical vascular receptor(s) may contribute to the increased severity of cardiovascular pathology in older persons. In vivo murine phage-display peptide library biopanning revealed a senescent decrease in cardiac microvascular binding of phage epitopes homologous to tumor necrosis factor-alpha (TNF-alpha), suggesting that its receptor(s) may be downregulated in older cardiac endothelial cells. Immunostaining demonstrated that TNF-receptor 1 (TNF-R1) density was significantly lower in the subendocardial endothelium of the aging murine heart. Functional studies confirmed the senescent dysregulation of TNF-alpha receptor pathways, demonstrating that TNF-alpha induced PDGF-B expression in cardiac microvascular endothelial cells of 4-mo-old, but not 24-mo-old, rats. Moreover, TNF-alpha mediated cardioprotective pathways were impaired in the aging heart. In young rat hearts, injection of TNF-alpha significantly reduced the extent of myocardial injury after coronary ligation: TNF-alpha, 7.9 +/- 1.9% left ventricular injury (n = 4) versus PBS, 16.2 +/- 7.9% (n = 10; P < 0.05). The addition of PDGF-AB did not augment the cardioprotective action of TNF-alpha. In myocardial infarctions of older hearts, however, TNF-alpha induced significant postcoronary occlusion mortality (TNF-alpha 80% vs. PBS 0%; n = 10 each, P < 0.05) that was reversed by the coadministration of PDGF-AB. Overall, these studies demonstrate that aging associated alterations in TNF-alpha receptor cardiac microvascular pathways may contribute to the increased cardiovasular pathology of the aging heart. Strategies targeted at restoring TNF-alpha receptor-mediated expression of PDGF-B may improve cardiac microvascular function and provide novel approaches for treatment and possible prevention of cardiovascular disease in older individuals. PMID- 12730064 TI - Antioxidant metabolism during acclimation of Begonia x erythrophylla to high light levels. AB - This study examined the influence of high light levels on antioxidant metabolism and the photosynthetic properties of Begonia x erythrophylla leaves. The pigment composition of shaded leaves and those developing in full sunlight was typical of shade- and sun-leaves, respectively. After 28 d in full sunlight, the preformed leaves of shade plants transferred to full sunlight (transferred-leaves) showed photo-bleaching with lower Chl (a + b) content and Chl a : Chl b ratios than shade-leaves, with Chl (a + b) : carotenoid ratios not significantly different. The variable/maximal fluorescence (Fv/Fm) of sun-leaves was not significantly different from that of shade-leaves, but transferred-leaves had reduced Fv : Fm ratios. Light response curves for the electron transport rate (ETR), the oxidation state of photosystem II (qP) and non-photochemical quenching (NPQ) showed significant differences between the three leaf types, with transferred leaves not able to acclimate completely to full sunlight, having lower ETR, qP and NPQ values at high light levels than sun-leaves. Transfer to full sunlight caused a rapid increase in H2O2 and lipid hyperoxides, and a slight increase in protein oxidation. Ascorbate and glutathione levels decreased rapidly, as did the size of the total glutathione pool and, in addition to the general oxidation of proteins, rapid decreases in both the initial and total activities of chloroplastic fructose-1,6-bisphosphatase and glyceraldehyde-3-phosphate dehydrogenase were observed. These results suggest that a more oxidizing cellular environment is the likely cause of the photo-bleaching observed upon transfer of shade-leaves to full sunlight. Acclimation of transferred-leaves to full sunlight involved gradual increases in the activities of enzymes involved in antioxidant metabolism, including superoxide dismutase, catalase, glutathione reductase, ascorbate peroxidase, dehydroascorbate reductase and monodehydroascorbate reductase, but the levels of these enzymes still remained at levels lower than those found in sun-leaves. PMID- 12730065 TI - A coupled model of photosynthesis, stomatal conductance and transpiration for a rose leaf (Rosa hybrida L.). AB - The following three models were combined to predict simultaneously photosynthesis, stomatal conductance, transpiration and leaf temperature of a rose leaf: the biochemical model of photosynthesis of Farquhar, von Caemmerer and Berry (1980, Planta 149: 78-90), the stomatal conductance model of Ball, Woodrow and Berry (In: Biggens J, ed. Progress in photosynthesis research. The Netherlands: Martinus Nijhoff Publishers), and an energy balance model. The photosynthetic parameters: maximum carboxylation rate, potential rate of electron transport and rate of triose phosphate utilization, and their temperature dependence were determined using gas exchange data of fully expanded, young, sunlit leaves. The stomatal conductance model was calibrated independently. Prediction of net photosynthesis by the coupled model agreed well with the validation data, but the model tended to underestimate rates of stomatal conductance and transpiration. The coupled model developed in this study can be used to assist growers making environmental control decisions in glasshouse production. PMID- 12730066 TI - Size-dependent growth and the development of inequality in maize, sunflower and soybean. AB - Links were investigated between allometry of plant growth and dynamics of size structure of well-fertilized, irrigated crops of soybean (Glycine max L.), sunflower (Helianthus annuus L.) and maize (Zea mays L.) grown at standard plant population densities (D), as in commercial crops (D = 30, 6 and 8.5 plants m-2, respectively), and at high densities (2D). Patterns of size-dependent growth of shoot and seed mass accumulation were distinctly different among species. In soybean and sunflower, non-linear relationships between size and subsequent growth led to strong hierarchical populations in terms of both shoot and seed biomass. Curvilinear (soybean) and sigmoid (sunflower) size-dependent growth determined strongly asymmetrical (soybean) and bimodal (sunflower) frequency distributions of shoot biomass indicating predominantly size asymmetrical competition among individuals. In comparison, a lower plant-to-plant variation coupled with a typical linear allometry of growth to plant size indicated symmetrical two-sided plant interference in maize. Despite the weak development of hierarchies in shoot biomass, a strong inequality in reproductive output developed in crowded populations of maize indicating an apparent breakage of reproductive allometry. PMID- 12730068 TI - Evidence for ovarian self-incompatibility as a cause of self-sterility in the relictual woody angiosperm, Pseudowintera axillaris (Winteraceae). AB - Species within the genus Pseudowintera exhibit high rates of self-sterility. Self sterility in the genus has been previously posited-but not confirmed-to be the result of late-acting ovarian self-incompatibility (OSI) functioning within nucellar tissue of the ovule to prevent self pollen tubes from entering the embryo sac. Structural and functional aspects of pollen-carpel interactions and early seed development following cross- and self-pollination were investigated in P. axillaris to determine the site, timing and possible mechanisms of self sterility. No significant differences were observed between pollen tube growth, ovule penetration and double fertilization following cross- and self-pollination. Pollen tubes exhibited phasic growth in an extracellular matrix composed of proteins and carbohydrates, as well as arabinogalactans/arabinogalactan proteins. A uniform failure in embryo sac development prior to division of the zygote was apparent within 15 d following double fertilization by self gametes. Results indicate that SI mechanisms in P. axillaris do not prevent double fertilization from occurring. Instead, mechanisms of self-sterility affect post-zygotic development of the embryo sac. Although self-sterility may be attributed to inbreeding depression, given the post-zygotic nature of failure in embryo sac development, the possibility of late-acting OSI is discussed. PMID- 12730067 TI - Tuber storage proteins. AB - A wide range of plants are grown for their edible tubers, but five species together account for almost 90 % of the total world production. These are potato (Solanum tuberosum), cassava (Manihot esculenta), sweet potato (Ipomoea batatus), yams (Dioscorea spp.) and taro (Colocasia, Cyrtosperma and Xanthosoma spp.). All of these, except cassava, contain groups of storage proteins, but these differ in the biological properties and evolutionary relationships. Thus, patatin from potato exhibits activity as an acylhydrolase and esterase, sporamin from sweet potato is an inhibitor of trypsin, and dioscorin from yam is a carbonic anhydrase. Both sporamin and dioscorin also exhibit antioxidant and radical scavenging activity. Taro differs from the other three crops in that it contains two major types of storage protein: a trypsin inhibitor related to sporamin and a mannose-binding lectin. These characteristics indicate that tuber storage proteins have evolved independently in different species, which contrasts with the highly conserved families of storage proteins present in seeds. Furthermore, all exhibit biological activities which could contribute to resistance to pests, pathogens or abiotic stresses, indicating that they may have dual roles in the tubers. PMID- 12730069 TI - Histological study of post-pollination events in Spathodea campanulata beauv. (Bignoniaceae), a species with late-acting self-incompatibility. AB - The reproductive biology of Spathodea campanulata was investigated by means of hand-pollination experiments, observations of pollen tube growth using fluorescence microscopy, and serial sections of ovules in selfed and crossed pistils. Only cross-pollinated flowers developed fruits, and all selfed flowers abscised within 3-4 d. However, self pollen tubes grew successfully to the ovary, penetrating and fertilizing the majority of ovules by 48 h, indicating that S. campanulata is a species with late-acting self-incompatibility. The incidences of ovule penetration, fertilization and endosperm initiation were all significantly slower in selfed vs. crossed pistils, although no other signs of malfunctioning were detected. The possible role of such slow self pollen tube effectiveness as a recognition event is discussed within the context of the slow but not entirely suppressed self pollen tube growth reported for some species with conventional homomorphic self-incompatibility. PMID- 12730070 TI - Control of kernel weight and kernel water relations by post-flowering source-sink ratio in maize. AB - The maize (Zea mays L.) kernel undergoes large changes in water content during its development. Whether such changes regulate the pattern of kernel development or are simply a consequence of it has not yet been established because other factors, such as assimilate supply, can also affect the rate and duration of kernel growth. This study was conducted to determine whether variation in kernel weight (KW) in response to source-sink treatments is mediated by a change in kernel water relations. Two hybrids were sown at three stand densities (one, eight and 18 plants m-2), and kernel numbers were restricted to control the post flowering source-sink ratio within each stand density. Kernel development and water relations [water content, water potential (psiw), osmotic potential (psis) and turgor] were monitored throughout grain filling. Final KW varied from 253 to 372 mg per kernel in response to source-sink treatments. For both genotypes, variation in KW was a result of a change in kernel growth rate (r2 = 0.91; P < 0.001) and not in the duration of kernel filling. Final KW was closely correlated with maximum kernel water content (r2 = 0.94; P < 0.001) achieved during rapid dry matter accumulation. However, variation in KW was not reflected in kernel water status parameters (psiw, psis or turgor), which remained fairly stable across treatments. These results indicate that maximum water content provides an easily quantifiable measure of kernel sink capacity in maize. Kernel water status parameters may affect the duration of grain filling, but have no discernible impact on kernel growth rate. PMID- 12730071 TI - Some quantitative relationships between leaf area index and canopy nitrogen content and distribution. AB - In a previous study (Yin et al. 2000. Annals of Botany 85: 579-585), a generic logarithmic equation for leaf area index (L) in relation to canopy nitrogen content (N) was developed: L=(1/ktn)1n(1+ktnN/nb). The equation has two parameters: the minimum leaf nitrogen required to support photosynthesis (nb), and the leaf nitrogen extinction coefficient (ktn). Relative to nb, there is less information in the literature regarding the variation of ktn. We therefore derived an equation to theoretically estimate the value of ktn. The predicted profile of leaf nitrogen in a canopy using this theoretically estimated value of ktn is slightly more uniform than the profile predicted by the optimum nitrogen distribution that maximizes canopy photosynthesis. Relative to the optimum profile, the predicted profile is somewhat closer to the observed one. Based on the L-N logarithmic equation and the theoretical ktn value, we further quantified early leaf area development of a canopy in relation to nitrogen using simulation analysis. In general, there are two types of relations between L and N, which hold for canopies at different developmental phases. For a fully developed canopy where the lowest leaves are senescing due to nitrogen shortage, the relationship between L and N is described well by the logarithmic model above. For a young, unclosed canopy (i.e. L < 1.0), the relation between L and N is nearly linear. This linearity is virtually the special case of the logarithmic model when applied to a young canopy where its total nitrogen content approaches zero and the amount of nitrogen in its lowest leaves is well above nb. The expected patterns of the L-N relationship are discussed for the phase of transition from young to fully developed canopies. PMID- 12730072 TI - Increase in leaf mass per area benefits plant growth at elevated CO2 concentration. AB - An increase in leaf mass per area (MLA) of plants grown at elevated [CO2] is often accompanied by accumulation of non-structural carbohydrates, and has been considered to be a response resulting from source-sink imbalance. We hypothesized that the increase in MLA benefits plants by increasing the net assimilation rate through maintaining a high leaf nitrogen content per area (NLA). To test this hypothesis, Polygonum cuspidatum was grown at ambient (370 micro mol mol-1) and elevated (700 micro mol mol-1) [CO2] with three levels of N supply. Elevated [CO2] significantly increased MLA with smaller effects on NLA and leaf mass ratio (fLM). The effect of change in MLA on plant growth was investigated by the sensitivity analysis: MLA values observed at ambient and elevated [CO2] were substituted into a steady-state growth model to calculate the relative growth rate (R). At ambient [CO2], substitution of a high MLA (observed at elevated [CO2]) did not increase R, compared with R for a low MLA (observed at ambient [CO2]), whereas at elevated [CO2] the high MLA always increased R compared with R at the low MLA. These results suggest that the increase in MLA contributes to growth enhancement under elevated [CO2]. The optimal combination of fLM and MLA to maximize R was determined for different [CO2] and N availabilities. The optimal fLM was nearly constant, while the optimal MLA increased at elevated [CO2], and decreased at higher N availabilities. The changes in fLM of actual plants may compensate for the limited plasticity of MLA. PMID- 12730075 TI - Food hoarding is increased by food deprivation and decreased by leptin treatment in Syrian hamsters. AB - Compensatory increases in food intake are commonly observed after a period of food deprivation in many species, including laboratory rats and mice. Thus it is interesting that Syrian hamsters fail to increase food intake after a period of food deprivation, despite a fall in plasma leptin concentrations similar to those seen in food-deprived rats and mice. In previous laboratory studies, food deprived Syrian hamsters increased the amount of food hoarded. We hypothesized that leptin treatment during food deprivation would attenuate food-deprivation induced increases in hoarding. Baseline levels of hoarding were bimodally distributed, with no hamsters showing intermediate levels of hoarding. Both high (HH) and low hoarding (LH) hamsters were included in each experimental group. Fifty-six male hamsters were either food deprived or given ad libitum access to food for 48 h. One-half of each group received intraperitoneal injections of leptin (4 mg/kg) or vehicle every 12 h during the food-deprivation period. Within the HH group, the hoarding score increased significantly in food-deprived but not fed hamsters (P < 0.05). Leptin treatment significantly decreased hoarding in the food-deprived HH hamsters (P < 0.05). The LH hamsters did not increase hoarding regardless of whether they were food deprived or had ad libitum access to food. These results are consistent with the idea that HH hamsters respond to energetic challenges at least in part by changing their hoarding behavior and that leptin might be one factor that mediates this response. PMID- 12730073 TI - Interleukin-11 and interleukin-6 protect cultured human endothelial cells from H2O2-induced cell death. AB - Acute lung injury is a frequent and treatment-limiting consequence of therapy with 100% oxygen. Previous studies have determined that both interleukin (IL)-6 and IL-11 are protective in oxygen toxicity. This protection was associated with markedly diminished alveolar-capillary protein leak, endothelial and epithelial membrane injury, lipid peroxidation, and pulmonary neutrophil recruitment. Hyperoxia also caused cell death with DNA fragmentation in the lungs of transgene (-) animals, and both IL-6 and IL-11 markedly diminished this cell death response. However, the mechanism(s) by which these cytokines protect cells from death is unclear. In the present study, we characterized the effects of H2O2 on subconfluent human umbilical vein endothelial cell (HUVEC) and human pulmonary microvascular endothelial cell (HPMEC) cultures. We found that preincubation of HUVEC cultures with either IL-6 or IL-11 diminished H2O2 (1.0 mM)-induced cell death. Similar effects were noted with HPMEC showing that this effect is not HUVEC-specific. The protective effects of both IL-6 and IL-11 were not associated with any changes in antioxidants and were decreased by approximately 80% in the presence of U0126, a specific inhibitor of MEK-1-dependent pathways. The cytoprotective effects of IL-11 and IL-6 were also completely eliminated in STAT3 dominant-negative transduced HUVEC cultures. These studies demonstrate that IL-6 and IL-11 both confer cytoprotective effects that diminish oxidant-mediated endothelial cell injury. They also demonstrate that this protection is mediated, at least in part, by a STAT3 and MEK-1-dependent specific signal transduction pathway(s). PMID- 12730074 TI - Neutrophil adhesion molecule expression in the developing neonatal rat exposed to hyperoxia. AB - Neonatal rats have an increased tolerance to hyperoxia, which is associated with a diminished pulmonary inflammatory response compared with adults. To investigate this differing response, expression of the neutrophil adhesion molecules, L selectin and CD18, and levels of soluble L-selectin, were examined using flow cytometry and sandwich enzyme-linked immunosorbent assay on air-exposed neonatal rat neutrophils at 0-24 and 72 h and 7, 10, 14, and 21 d of age compared with the adult and after exposure to hyperoxia (>/= 98% O2) for 56 h in adults and for 72 h and 7 d in neonates. Expression of L-selectin in 0-24-h neonates was similar to adults, but was significantly lower than adults at 72 h and 7 d (P = 0.011). Soluble L-selectin levels were significantly higher than those in adults in the 0 24- and 72-h neonates (P < 0.001). CD18 expression in unstimulated and activated neutrophils of neonatal rats was higher at 0-24 h than in the adult (P < 0.001), but thereafter did not differ from adults. After hyperoxic exposure, L-selectin did not differ between the exposure groups but soluble L-selectin tended to increase in neonates after 7 d of O2 exposure Finally, CD18 was significantly higher after hyperoxic exposure of the adult (P = 0.008), but did not change with oxygen exposure in the neonate. Based on these findings, we speculate that differences between neonatal and adult rats in expression of L-selectin may contribute to delayed oxygen toxicity in neonatal rats. PMID- 12730076 TI - Changes in brain glycogen after sleep deprivation vary with genotype. AB - Sleep has been functionally implicated in brain energy homeostasis in that it could serve to replenish brain energy stores that become depleted while awake. Sleep deprivation (SD) should therefore lower brain glycogen content. We tested this hypothesis by sleep depriving mice of three inbred strains, i.e., AKR/J (AK), DBA/2J (D2), and C57BL/6J (B6), that differ greatly in their sleep regulation. After a 6-h SD, these mice and their controls were killed by microwave irradiation, and glycogen and glucose were quantified in the cerebral cortex, brain stem, and cerebellum. After SD, both measures significantly increased by approximately 40% in the cortex of B6 mice, while glycogen significantly decreased by 20-38% in brain stem and cerebellum of AK and D2 mice. In contrast, after SD, glucose content increased in all three structures in AK mice and did not change in D2 mice. The increase in glycogen after SD in B6 mice persisted under conditions of food deprivation that, by itself, lowered cortical glycogen. Furthermore, the strains that differ most in their compensatory response to sleep loss, i.e., AK and D2, did not differ in their glycogen response. Thus glycogen content per se is an unlikely end point of sleep's functional role in brain energy homeostasis. PMID- 12730077 TI - Beta-adrenergic agonists inhibit corticosteroid-induced apoptosis of airway epithelial cells. AB - Airway epithelial damage is a feature of persistent asthma. Treatment with inhaled and oral corticosteroids may suppress inflammation and gain clinical control despite continued epithelial damage. We have previously demonstrated that corticosteroids elicit apoptosis of airway epithelial cells in culture. beta Adrenergic receptor agonists are commonly used in asthma therapy and can inhibit corticosteroid-induced apoptosis of eosinophils. We tested the hypothesis that beta-adrenergic agonists would inhibit corticosteroid-induced airway epithelial cell apoptosis in cultured primary airway epithelial cells and in the cell line 1HAEo-. Albuterol treatment inhibited dexamethasone-induced apoptosis completely but did not inhibit apoptosis induced by Fas receptor activation. The protective effect of albuterol was duplicated by two different analogs of protein kinase A. The protective effect was not associated with increased translocation of the glucocorticoid receptor to the nucleus nor with changes in glucocorticoid receptor-mediated transcriptional activation or repression. We demonstrate that beta-adrenergic agonists can inhibit corticosteroid-induced apoptosis but not apoptosis induced by Fas activation. These data suggest that one potential deleterious effect of corticosteroid therapy in asthma can be prevented by concomitant beta-adrenergic agonist treatment. PMID- 12730078 TI - Epinephrine promotes pulmonary angiitis: evidence for a beta1-adrenoreceptor mediated mechanism. AB - Epinephrine (Epi) increases lymphocyte traffic to lung. We investigated whether Epi also modulates pulmonary cell-mediated immune responses in vivo. C57BL/6 mice were immunized with hen-egg lysozyme (HEL) on day 0, challenged with HEL intratracheally at day 12, and killed at day 15. Mice received Epi (0.5 mg/kg) subcutaneously during the sensitization phase, days 1-7 (Epi-SP), or the effector phase, days 12-14 (Epi-EP); controls received saline subcutaneously. Epi-SP mice showed increased airway inflammation (P < 0.03) and pulmonary angiitis (P < 0.04) characterized by endothelialitis and subendothelial fibrin deposition. Macrophages and granulocytes were increased in perivascular cuffs in situ (P < 0.001). CD3+ lymphocytes increased in the bronchoalveolar lavage fluid, whereas NK1.1+ and CD4+CD25+ lymphocytes decreased (all P < 0.05). Atenolol, a selective beta1-adrenoreceptor (AR) antagonist, inhibited the increased vascular and airway inflammation and the reduction in CD4+CD25+ lymphocytes (all P < 0.05) yielded by Epi, whereas all alpha/beta-AR blockers inhibited airway inflammation. We conclude that Epi-EP selectively promotes vascular inflammation in vivo via a beta1-receptor-mediated mechanism. PMID- 12730079 TI - Elevated levels of plasminogen activator inhibitor-1 in pulmonary edema fluid are associated with mortality in acute lung injury. AB - The alveolar fibrinolytic system is altered in acute lung injury (ALI). Levels of the fibrinolytic protease inhibitor, plasminogen activator inhibitor-1 (PAI-1), are too low in bronchoalveolar lavage to address its prognostic significance. This study was performed to assess whether PAI-1 antigen in undiluted pulmonary edema fluid levels can identify patients with ALI and predict their outcome. PAI 1 antigen levels in both plasma and edema fluid were higher in ALI compared with hydrostatic edema, and edema fluid PAI-1 values identified those with ALI with high sensitivity and specificity. Both the high plasma and edema fluid PAI-1 antigen values were associated with a higher mortality rate and fewer days of unassisted ventilation in patients with ALI. Differences in PAI-1 activity were concordant with levels of PAI-1 antigen. Although the fibrin-derived alveolar D dimer levels were strikingly similar in both groups, ALI patients had a higher relative proportion of D-monomer. In conclusion, PAI-1 levels in edema fluid and plasma identify those with ALI that have a poor prognosis. The data indicate that fibrin turnover in early ALI is a consequence of a rapid fibrinogen influx and fractional fibrinolytic inhibition. PMID- 12730080 TI - Modulation of ion conductance and active transport by TGF-beta 1 in alveolar epithelial cell monolayers. AB - Transforming growth factor-beta1 (TGF-beta 1) may be a critical mediator of lung injury and subsequent remodeling during recovery. We evaluated the effects of TGF beta 1 on the permeability and active ion transport properties of alveolar epithelial cell monolayers. Rat alveolar type II cells plated on polycarbonate filters in defined serum-free medium form confluent monolayers and acquire the phenotypic characteristics of alveolar type I cells. Exposure to TGF-beta 1 (0.1 100 pM) from day 0 resulted in a concentration- and time-dependent decrease in transepithelial resistance (Rt) and increase in short-circuit current (Isc). Apical amiloride or basolateral ouabain on day 6 inhibited Isc by 80 and 100%, respectively. Concurrent increases in expression of Na+-K+-ATPase alpha 1- and beta 1-subunits were observed in TGF-beta 1-treated monolayers. No change in the alpha-subunit of the rat epithelial sodium channel (alpha-rENaC) was seen. Exposure of confluent monolayers to TGF-beta 1 from day 4 resulted in an initial decrease in Rt within 6 h, followed by an increase in Isc over 72-96 h. These results demonstrate that TGF-beta 1 modulates ion conductance and active transport characteristics of the alveolar epithelium, associated with increased Na+-K+-ATPase, but without a change in alpha-rENaC. PMID- 12730081 TI - Involvement of reactive oxygen species in the metabolic pathways triggered by diesel exhaust particles in human airway epithelial cells. AB - Diesel exhaust particles (DEP) induce a proinflammatory response in human bronchial epithelial cells (16HBE) characterized by the release of proinflammatory cytokines after activation of transduction pathways involving MAPK and the transcription factor NF-kappaB. Because cellular effects induced by DEP are prevented by antioxidants, they could be mediated by reactive oxygen species (ROS). Using fluorescent probes, we detected ROS production in bronchial and nasal epithelial cells exposed to native DEP, organic extracts of DEP (OE DEP), or several polyaromatic hydrocarbons. Carbon black particles mimicking the inorganic part of DEP did not increase ROS production. DEP and OE-DEP also induced the expression of genes for phase I [cytochrome P-450 1A1 (CYP1A1)] and phase II [NADPH quinone oxidoreductase-1 (NQO-1)] xenobiotic metabolization enzymes, suggesting that DEP-adsorbed organic compounds become bioavailable, activate transcription, and are metabolized since the CYP1A1 enzymatic activity is increased. Because NQO-1 gene induction is reduced by antioxidants, it could be related to the ROS generated by DEP, most likely through the activation of the stress-sensitive Nrf2 transcription factor. Indeed, DEP induced the translocation of Nrf2 to the nucleus and increased protein nuclear binding to the antioxidant responsive element. In conclusion, we show that DEP-organic compounds generate an oxidative stress, activate the Nrf2 transcription factor, and increase the expression of genes for phase I and II metabolization enzymes. PMID- 12730082 TI - Multiaxial mechanical behavior of biological materials. AB - For native and engineered biological tissues, there exist many physiological, surgical, and medical device applications where multiaxial material characterization and modeling is required. Because biological tissues and many biocompatible elastomers are incompressible, planar biaxial testing allows for a two-dimensional (2-D) stress-state that can be used to fully characterize their three-dimensional (3-D) mechanical properties. Biological tissues exhibit complex mechanical behaviors not easily accounted for in classic elastomeric constitutive models. Accounting for these behaviors by careful experimental evaluation and formulation of constitutive models continues to be a challenging area in biomechanical modeling and simulation. The focus of this review is to describe the application of multiaxial testing techniques to soft tissues and their relation to modern biomechanical constitutive theories. PMID- 12730083 TI - Blood vessel constitutive models-1995-2002. AB - Knowledge of blood vessel mechanical properties is fundamental to the understanding of vascular function in health and disease. Analytic results can help physicians in the clinic, both in designing and in choosing appropriate therapies. Understanding the mechanical response of blood vessels to physiologic loads is necessary before ideal therapeutic solutions can be realized. For this reason, blood vessel constitutive models are needed. This article provides a critical review of recent blood vessel constitutive models, starting with a brief overview of the structure and function of arteries and veins, followed by a discussion of experimental techniques used in the characterization of material properties. Current models are classified by type, including pseudoelastic, randomly elastic, poroelastic, and viscoelastic. Comparisons are presented between the various models and existing experimental data. Applications of blood vessel constitutive models are also briefly presented, followed by the identification of future directions in research. PMID- 12730084 TI - Development of a genotype 325-specific proCPU/TAFI ELISA. AB - OBJECTIVE: A Thr/Ile polymorphism at position 325 in the coding region of proCPU has been reported. Immunological assays, fully characterized (including genotype dependency), are required for the quantitation of proCPU levels. METHODS AND RESULTS: We have generated a panel of monoclonal antibodies against human, plasma derived proCPU. Two combinations exhibiting distinct reactivities were selected for measurement of proCPU in plasma. T12D11/T28G6-HRP yielded values of 10.1+/ 3.1 microg/mL (mean+/-SD, n=86; normal donors), and T32F6/T9G12-HRP yielded values of 5.4+/-3.0 microg/mL. Grouping according to the 325 genotype demonstrated that T12D11/T28G6-HRP was independent to this polymorphism whereas T32F6/T9G12-HRP revealed a complete lack of reactivity with the Ile/Ile genotype (ie, 0.0+/-0.0, 4.2+/-1.7, and 7.3+/-2.9 microg/mL for the Ile/Ile, Ile/Thr, and Thr/Thr isoforms, respectively). Commercially available antigen assays appeared to be partially dependent on the 325 genotype (eg, 44+/-8.9% and 100+/-30% for the Ile/Ile and Thr/Thr isoforms, respectively). CONCLUSIONS: Our data demonstrate that great care should be taken when evaluating proCPU antigen values as a putative causative agent or as a diagnostic risk marker for cardiovascular events. PMID- 12730085 TI - Effect of oral and transdermal estrogen replacement therapy on hemostatic variables associated with venous thrombosis: a randomized, placebo-controlled study in postmenopausal women. AB - OBJECTIVE: The purpose of this study was to investigate whether the effect of transdermal estrogen therapy in postmenopausal women differs from that of oral therapy with regard to resistance to activated protein C (APC), an important risk factor for venous thrombosis, and levels of related proteins, such as protein S, protein C, and prothrombin. METHODS AND RESULTS: In a randomized, double-blind, placebo-controlled study, 152 healthy hysterectomized postmenopausal women received daily either placebo (n=49), transdermal 17beta-estradiol (E2) 50 microg (tE2 group, n=33), oral E2 1 mg (oE2 group, n=37), or oral E2 1 mg combined with gestodene 25 microg (oE2+G group, n=33) for 13 28-day treatment cycles, followed by 4 cycles of placebo for each group. Plasma samples were collected at baseline and in cycles 4, 13, and 17. In cycle 13, significant increases versus baseline and placebo were found in normalized APC sensitivity ratios (nAPCsr) in all treated groups (tE2, +26.9%; oE2, +102.7%; oE2+G, +69.9%). Increases in nAPCsr were significantly higher in the oral treatment groups than in the tE2 group. In addition, compared with baseline and placebo, after 13 cycles, decreases were observed in total protein S (tE2, -4.1%; oE2, -7.9%; oE2+G, -5.8%), free protein S (tE2, -7.1%; oE2, -8.4%; oE2+G, -5.2%), and protein C in the oE2+G group ( 6.4%), but these changes did not explain the increase in nAPCsr. Changes in prothrombin were small and also did not affect the nAPCsr. CONCLUSIONS: Increases were observed in resistance to APC, which were more pronounced in the oral treatment groups than in the transdermal group. The increase in resistance to APC was not explained by changes in protein S, protein C, or prothrombin and may contribute to the increased incidence of venous thrombosis in users of hormone replacement therapy. PMID- 12730086 TI - Amino acid differences in the deduced 5-lipoxygenase sequence of CAST atherosclerosis-resistance mice confer impaired activity when introduced into the human ortholog. AB - OBJECTIVE: The mouse strain CON6, which was generated by breeding athero resistant CAST mice into an athero-susceptible B6 background, exhibits almost complete resistance to atherosclerosis. An athero-resistance gene cluster has been localized at the central region of chromosome 6, and among the candidate genes of this locus, the 5-lipoxygenase has attracted particular attention because of its involvement in the biosynthesis of proinflammatory leukotrienes. Comparison of 5-lipoxygenase genomic sequences of B6 and CON6 mice indicated 2 conserved amino acid exchanges in the CON6 animals, but the functional impact of these mutations has not been defined. METHODS AND RESULTS: We analyzed the functionality of these amino acid exchanges relative to essential catalytic properties (specific activity, substrate affinity, and reaction specificity) and found that these mutations confer an impaired lipoxygenase and leukotriene A4 synthase activity when introduced into the human enzyme. In contrast, substrate affinity, enantiomer selectivity, and positional specificity remained unchanged. CONCLUSIONS: These data are consistent with the possibility that naturally occurring conservative mutations in the coding region of the murine 5 lipoxygenase gene can significantly affect enzyme activity and that this loss of function may be involved in CAST/CON6 athero-resistance. PMID- 12730087 TI - Osteopontin deficiency attenuates atherosclerosis in female apolipoprotein E deficient mice. AB - OBJECTIVE: Osteopontin (OPN), a noncollagenous adhesive protein, is implicated in atherosclerosis, in which macrophages within atherosclerotic plaques express OPN. However, it is not known whether the elevated OPN expression is a cause or result of atherosclerosis. METHODS AND RESULTS: We generated mice that lacked OPN and crossed them with apolipoprotein (apo) E-deficient mice and analyzed these mice with a mixed C57BL/6x129 background after 36 weeks on a normal chow diet. In female mice, OP+/-E-/- and OP-/-E-/- mice had significantly smaller atherosclerotic and inflammatory lesions compared with OP+/+E-/- mice, and that was reflected by smaller area of MOMA-2-positive staining. In male mice, however, there was no significant difference in the atherosclerosis lesion areas among 3 genotypes. In both OP-/-E-/- and OP+/+E-/- mice, typical atherosclerotic lesions were detected, which include necrotic core, foamy cell collections, and cholesterol clefts. However, we found that vascular mineral-deposited areas in 60 week-old male OP-/-E-/- mice were significantly increased compared with those in OP+/+E-/- male mice. CONCLUSIONS: These results suggest that OPN plays a promoting effect in atherosclerosis and inhibitory effect in vascular calcification. The suppression of OPN expression in females should be considered a therapeutic possibility in atherosclerosis. PMID- 12730088 TI - Effects of cyclooxygenases inhibitors on vasoactive prostanoids and thrombin generation at the site of microvascular injury in healthy men. AB - OBJECTIVE: Balance between vasoactive prostanoids that contribute to homeostasis of the circulatory system can be affected by cyclooxygenases inhibitors. Results of a recent large clinical trial show that myocardial infarction was more frequent among patients with rheumatoid arthritis treated with the selective cyclooxygenase-2 inhibitor rofecoxib compared with those treated with naproxen. Whether this difference was attributable to deleterious cardiovascular effects of rofecoxib or cardioprotective effects of naproxen has not been determined. We tested the hypothesis that naproxen, contrary to rofecoxib, exerts antithrombotic effects. METHODS AND RESULTS: Forty-five healthy men were randomized to receive a 7-day treatment with rofecoxib (50 mg/d), naproxen (1000 mg/d), aspirin (75 mg/d), or diclofenac (150 mg/d). Formation of thromboxane, prostacyclin, and thrombin in the bleeding-time blood at the site of standardized microvascular injury was assessed before and after treatment. Naproxen, like aspirin, caused significant reduction of both thromboxane and prostacyclin, whereas diclofenac depressed prostacyclin synthesis but had no effect on tromboxane formation. Naproxen and aspirin significantly suppressed thrombin generation. Diclofenac showed a similar tendency, which did not reach statistical significance. Rofecoxib had no effect on any variables measured. CONCLUSIONS: In healthy men, naproxen exerts an antithrombotic effect at least as potent as aspirin, whereas rofecoxib does not affect hemostatic balance. PMID- 12730089 TI - Increased serum levels of heat shock protein 70 are associated with low risk of coronary artery disease. AB - OBJECTIVE: Previous studies suggest that heat shock protein (HSP) 60 has a contributory role in atherosclerosis development. We examined whether circulating HSP70 protein and anti-HSP70 antibodies are associated with coronary artery disease (CAD). METHODS AND RESULTS: Blood samples from 421 patients (62% men, mean age 57 years) evaluated for CAD by coronary angiography were tested. Serum HSP70 was detectable in 67% of study subjects with levels ranging from 0.2 to 27.1 ng/mL (mean, 1.08; median, 0.5). HSP70 levels were higher in non-CAD patients than CAD patients (median, 0.72 versus 0.34; P=0.0006). Individuals with HSP70 levels above the median (0.5 ng/mL) had half the risk of CAD than individuals with levels below the median (adjusted odds ratio, 0.52; 95% confidence limit, 0.32 to 0.86). The association of high HSP70 levels with low CAD risk was independent of traditional CAD risk factors (P=0.011). Disease severity (number of diseased vessels) was also inversely associated with HSP70 protein levels (P=0.010). The adjusted odds ratio of having multivessel disease for patients with high HSP70 protein levels was 0.54 (95% confidence limit, 0.36 to 0.81). In contrast, no association between anti-HSP70 IgG seropositivity and the prevalence of CAD was found (P=0.916). CONCLUSIONS: These data provide the first evidence that high levels of human HSP70 are associated with the low CAD risk, probably through its multiple protective effects on a cell's response to stress. PMID- 12730090 TI - Genes, environment, and cardiovascular disease. AB - In this essay, we call to attention what every medical researcher knows about the etiology of cardiovascular disease but most deny, or choose to ignore, when designing, carrying out, and reporting genetic studies. Medical research is entering an era of synthesis that will take advantage of the successes of reductionism over the past decade in defining and describing human genome variations. Meaningful insights into the role of such variation requires a biological model of genome-phenotype relationships that incorporates interactions between subsets of possible genetic and environmental agents as causations in particular contexts indexed by time and space. We make recommendations for what needs to be done to cope with these complexities. PMID- 12730091 TI - Phosphatidylinositol 3-kinase and calcium-activated transcription pathways are required for VLDL-induced smooth muscle cell proliferation. AB - Little is known regarding the molecular mechanisms of atherogenicity of triglyceride-rich lipoproteins such as very low-density lipoproteins (VLDLs). We examined the effect of VLDL on proliferation of rat aortic smooth muscle cells, intracellular Ca2+ handling, and activity of cAMP-responsive element binding protein (CREB) and nuclear factor of activated T cells (NFAT) transcription factors. VLDL, isolated from human serum, dose- and time-dependently promoted proliferation. After 4 hours of exposure to VLDL (0.15 g/L proteins), the caffeine-induced Ca2+ release was inhibited and the IP3-sensitive Ca2+ release induced by ATP (10 micromol/L) was markedly prolonged. In quiescent cells, CREB was phosphorylated (pCREB) and NFAT was present in the cytosol, whereas in cells exposed to VLDL for 4 to 24 hours, pCREB disappeared and NFAT was translocated to the nucleus. VLDL-induced NFAT translocation and proliferation were blocked by cyclosporin A and LY294002 involving calcineurin and phosphatidylinositol 3 kinase (PI3K) pathways. Indeed, VLDLs rapidly phosphorylate protein kinase B and glycogen synthase kinase-3beta in a PI3K-dependent way. These results provide the first evidence that VLDLs induce smooth muscle cell proliferation by activating the PI3K pathway and nuclear NFAT translocation. Blockade of the Ca2+-induced Ca2+ release mechanism and dephosphorylation of pCREB contribute but were not sufficient to induce a proliferating phenotype. PMID- 12730093 TI - Mechanisms of delayed electrical uncoupling induced by ischemic preconditioning. AB - Electrical uncoupling of cardiac myocytes during ischemia is delayed by ischemic preconditioning. This presumably adaptive response may limit development of arrhythmia substrates. To elucidate responsible mechanisms, we studied isolated, perfused rat hearts subjected to a standard preconditioning protocol of 3 cycles of 3 minutes of global no-flow ischemia each followed by 5 minutes of reperfusion before a 30-minute interval of ischemia. Changes in coupling were monitored by measuring whole-tissue resistance. Changes in phosphorylation and subcellular distribution of connexin43 (Cx43) were defined by quantitative immunoblotting and confocal microscopy. Preconditioning caused a 34% decrease in the maximal rate of uncoupling and delayed the time to plateau in uncoupling. Dephosphorylation of Cx43, known to occur during uncoupling induced by ischemia, was dramatically decreased in preconditioned hearts. Translocation of Cx43 from gap junctions to the cytosol, also known to occur during ischemia, was reduced by >5-fold in preconditioned hearts. The KATP channel blockers glybenclamide and 5 hydroxydecanoate prevented these effects in preconditioned hearts, whereas the KATP channel agonist diazoxide mimicked these effects in nonpreconditioned hearts. Intracellular translocation of Cx43 was blocked, but Cx43 dephosphorylation was not blocked during ischemia in preconditioned hearts treated with the PKC inhibitors chelerythrine and calphostin C. Uncoupling during ischemia was accelerated by PKC and KATP channel inhibition. Thus, delayed uncoupling in preconditioned hearts is likely related to diminished dephosphorylation and intracellular redistribution of Cx43 during prolonged ischemia. Both of these effects are regulated by activation of KATP channels, whereas PKC plays a role in internalization of Cx43. PMID- 12730092 TI - Receptor tyrosine kinase Axl modulates the osteogenic differentiation of pericytes. AB - Vascular pericytes undergo osteogenic differentiation in vivo and in vitro and may, therefore, be involved in diseases involving ectopic calcification and osteogenesis. The purpose of this study was to identify factors that inhibit the entry of pericytes into this differentiation pathway. RNA was prepared from pericytes at confluence and after their osteogenic differentiation (mineralized nodules). Subtractive hybridization was conducted on polyA PCR-amplified RNA to isolate genes expressed by confluent pericytes that were downregulated in the mineralized nodules. The subtraction product was used to screen a pericyte cDNA library and one of the positive genes identified was Axl, the receptor tyrosine kinase. Northern and Western blotting confirmed that Axl was expressed by confluent cells and was downregulated in mineralized nodules. Western blot analysis demonstrated that confluent pericytes also secrete the Axl ligand, Gas6. Immunoprecipitation of confluent cell lysates with an anti-phosphotyrosine antibody followed by Western blotting using an anti-Axl antibody, demonstrated that Axl was active in confluent pericytes and that its activity could not be further enhanced by incubating the cells with recombinant Gas6. The addition of recombinant Axl-extracellular domain (ECD) to pericyte cultures inhibited the phosphorylation of Axl by endogenous Gas6 and enhanced the rate of nodule mineralization. These effects were inhibited by coincubation of pericytes with Axl-ECD and recombinant Gas6. Together these results demonstrate that activation of Axl inhibits the osteogenic differentiation of vascular pericytes. PMID- 12730094 TI - Leprdb diabetic mouse bone marrow cells inhibit skin wound vascularization but promote wound healing. AB - Bone marrow stem cells participate in tissue repair processes and may have roles in skin wound repair. Diabetes is characterized by delayed and poor wound healing, and type 1 diabetes seems to lead to stem cell dysfunction. Hence, stem cell dysfunction could contribute to poor healing, and stem cell-based therapies may be efficacious in diabetic wounds. We investigated the potential of exogenous stem cells to promote skin healing and possible effects of type 2 diabetes on stem cell function. Mouse bone marrow cells from nondiabetic and diabetic mice were enriched for putative stem cells and injected under skin wounds of nondiabetic or type 2 diabetic Leprdb mice. Using histology and morphometry, vascularization and healing in treated and untreated mice were analyzed. We anticipated a correlation between improved wound healing and vascularization, because therapies that increase tissue vascularization tend to enhance wound healing. Our data indicate that exogenous nondiabetic bone marrow-derived cells increase vascularization and improve wound healing in Leprdb mice but have little effect on nondiabetic controls. In contrast, Leprdb-derived marrow cells inhibit vascularization but promote wound healing in Leprdb mice. Thus, adult stem cell function may be impaired by type 2 diabetes; the ability to promote vascularization and wound healing are distinct functions of bone marrow cells; and neovascularization and wound healing may not be tightly coupled. Additionally, we observed little incorporation of injected cells into wound structures, suggesting that improved healing is mediated through mechanisms other than direct differentiation and incorporation of the cells. PMID- 12730096 TI - Physiological coupling of donor and host cardiomyocytes after cellular transplantation. AB - Cellular transplantation has emerged as a potential approach to treat diseased hearts. Although cell transplantation can affect global heart function, it is not known if this results directly via functional integration of donor myocytes or indirectly via enhanced revascularization and/or altered postinjury remodeling. To determine the degree to which donor cardiomyocytes are able to functionally integrate with the host myocardium, fetal transgenic cardiomyocytes expressing enhanced green fluorescent protein were transplanted into the hearts of nontransgenic adult mice. Two-photon molecular excitation laser scanning microscopy was then used to simultaneously image cellular calcium transients in donor and host cells within the intact recipient hearts. Calcium transients in the donor cardiomyocytes were synchronous with and had kinetics indistinguishable from those of neighboring host cardiomyocytes. These results strongly suggest that donor cardiomyocytes functionally couple with host cardiomyocytes and support the notion that transplanted cardiomyocytes can form a functional syncytium with the host myocardium. PMID- 12730095 TI - Impulse propagation in synthetic strands of neonatal cardiac myocytes with genetically reduced levels of connexin43. AB - Connexin43 (Cx43) is a major determinant of the electrical properties of the myocardium. Closure of gap junctions causes rapid slowing of propagation velocity (theta), but the precise effect of a reduction in Cx43 levels due to genetic manipulation has only partially been clarified. In this study, morphological and electrical properties of synthetic strands of cultured neonatal ventricular myocytes from Cx43+/+ (wild type, WT) and Cx+/- (heterozygote, HZ) mice were compared. Quantitative immunofluorescence analysis of Cx43 demonstrated a 43% reduction of Cx43 expression in the HZ versus WT mice. Cell dimensions, connectivity, and alignment were independent of genotype. Measurement of electrical properties by microelectrodes and optical mapping showed no differences in action potential amplitude or minimum diastolic potential between WT and HZ. However, maximal upstroke velocity of the transmembrane action potential, dV/dtmax, was increased and action potential duration was reduced in HZ versus WT. theta was similar in the two genotypes. Computer simulation of propagation and dV/dtmax showed a relatively small dependence of theta on gap junction coupling, thus explaining the lack of observed differences in theta between WT and HZ. Importantly, the simulations suggested that the difference in dV/dtmax is due to an upregulation of INa in HZ versus WT. Thus, heterozygote null mutation of Cx43 produces a complex electrical phenotype in synthetic strands that is characterized by both changes in ion channel function and cell-to cell coupling. The lack of changes in theta in this tissue is explained by the dominating role of myoplasmic resistance and the compensatory increase of dV/dtmax. PMID- 12730097 TI - Mature parasite-infected erythrocyte surface antigen (MESA) of Plasmodium falciparum binds to the 30-kDa domain of protein 4.1 in malaria-infected red blood cells. AB - The Plasmodium falciparum mature parasite-infected erythrocyte surface antigen (MESA) is exported from the parasite to the infected red blood cell (IRBC) membrane skeleton, where it binds to protein 4.1 (4.1R) via a 19-residue MESA sequence. Using purified RBC 4.1R and recombinant 4.1R fragments, we show MESA binds the 30-kDa region of RBC 4.1R, specifically to a 51-residue region encoded by exon 10 of the 4.1R gene. The 3D structure of this region reveals that the MESA binding site overlaps the region of 4.1R involved in the p55, glycophorin C, and 4.1R ternary complex. Further binding studies using p55, 4.1R, and MESA showed competition between p55 and MESA for 4.1R, implying that MESA bound at the IRBC membrane skeleton may modulate normal 4.1R and p55 interactions in vivo. Definition of minimal binding domains involved in critical protein interactions in IRBCs may aid the development of novel therapies for falciparum malaria. PMID- 12730098 TI - Microsatellite polymorphism in heme oxygenase-1 gene promoter is associated with susceptibility to oxidant-induced apoptosis in lymphoblastoid cell lines. AB - Heme oxygenase-1 (HO-1) confers cytoprotection against oxidative stress. A (GT)n dinucleotide repeat in the 5'-flanking region of human HO-1 gene shows length polymorphism, which was classified into S (< 27 GT), M (27-32 GT), and L alleles (>/= 33 GT). Polymorphism in the HO-1 gene promoter was shown to be associated with susceptibility to pulmonary emphysema and restenosis after angioplasty. However, the biologic mechanism underlying these associations is still unclear. To examine this issue, we established lymphoblastoid cell lines (LCLs) from subjects possessing S/S or L/L genotypes. HO-1 mRNA expressions and HO activities induced by oxidative stress were significantly higher in LCLs with S/S than those with L/L. Furthermore, LCLs with S/S were significantly more resistant to oxidant induced apoptosis than those with L/L. These findings suggested that the polymorphism of the HO-1 gene is associated with the strength of antiapoptotic effects of HO-1, resulting in an association with susceptibility to oxidative stress-mediated diseases. PMID- 12730099 TI - Integration of DAG signaling systems mediated by PKC-dependent phosphorylation of RasGRP3. AB - Members of the RasGRP family of Ras activators have C1 domains that bind diacylglycerol (DAG) and DAG analogs such as the tumor-promoting phorbol esters. RasGRP members could be responsible for some of the DAG signaling processes that have previously been attributed to protein kinase C (PKC). We found that RasGRP3 is selectively expressed in B cells, suggesting that RasGRP3 might function downstream of the B-cell receptor (BCR). Indeed, stimulation of Ramos B cells with the DAG analog phorbol ester myristate (PMA) results in the association of RasGRP3 with the membrane fraction. However, we also made the unexpected observation that RasGRP3 is phosphorylated, coincident with Ras activation after stimulation. When inhibitors of PKC are present, Ras activation is attenuated, and this attenuation correlates with an inhibition of RasGRP3 phosphorylation. RasGRP3 is phosphorylated in vitro by PKC-theta and PKC-beta2. When ectopically coexpressed in HEK-293 cells, a dominant-activated mutant of PKC-theta phosphorylates RasGRP3 and enhances Ras-Erk signaling. These results provide the first indication for a functional interaction between a RasGRP family member and a dissimilar DAG binding protein. A convergent DAG signaling system could be important in fine-tuning Ras signaling during B-cell development or during the humoral immune response. PMID- 12730100 TI - Synergistic effect of sphingosine 1-phosphate on thrombin-induced tissue factor expression in endothelial cells. AB - Sphingosine 1-phosphate (S1P), a bioactive lipid, is produced and stored in platelets and is released from activated platelets during blood coagulation activation. Thrombin, which is also generated during blood coagulation, has been shown to induce tissue factor (TF), the initiator of blood coagulation, in endothelial cells (ECs); however, the effect of S1P on this process is not evaluated. Here we demonstrated that S1P strongly potentiated thrombin-induced TF expression in ECs and that S1P itself did not induce TF expression. Among signaling lipids, platelet-activating factor slightly enhanced thrombin-induced TF expression; other lipids, including lysophosphatidic acid, lysophosphatidylcholine, sphingosine, and C2-ceramide exert no effect on TF expression. S1P enhanced TF expression at the transcriptional level, possibly via promoting the activation of transcription factors nuclear factor-kappaB (NF kappaB) and Egr-1. Thrombin weakly and S1P strongly activated extracellular signal-regulated kinase 1/2 (ERK1/2) mitogen-activated protein (MAP) kinase and, in the presence of both stimulants, enhanced and sustained activation of this kinase was observed. The ERK1/2-specific inhibitor PD98059 significantly inhibited enhanced TF expression induced by both stimulants but only weakly inhibited thrombin-induced TF expression, thus indicating the requirement of the ERK1/2 pathway in synergistic induction of TF expression. In addition, we found that thrombin and S1P rapidly up-regulated the expression of S1P receptors, endothelial differentiation gene-1 (EDG-1) and EDG-3, thereby suggesting that the effect of S1P on TF expression and other EC functions may be enhanced by thrombin and S1P itself. The present data reveal the synergistic effect of S1P on thrombin induced TF expression in ECs, which may promote further thrombin and S1P generation, thus propagating a positive feedback reaction. PMID- 12730101 TI - Retrovirus-mediated gene transfer in polyclonal T cells results in lower apoptosis and enhanced ex vivo cell expansion of CMV-reactive CD8 T cells as compared with EBV-reactive CD8 T cells. AB - To modulate alloreactivity after hematopoietic stem cell transplantation, "suicide" gene-modified donor T cells (GMCs) have been administered with an allogeneic T-cell-depleted marrow graft. We previously demonstrated that such GMCs, generated after CD3 activation, retrovirus-mediated transduction, and G418 selection, had an impaired Epstein-Barr virus (EBV) reactivity, likely to result in an altered control of EBV-induced lymphoproliferative disease. To further characterize the antiviral potential of GMCs, we compared the frequencies of cytomegalovirus (CMV)-specific CD8+ T (CMV-T) cells and EBV-specific CD8+ T (EBV T) cells within GMCs from CMV- and EBV-double seropositive donors. Unlike anti EBV responses, the anti-CMV responses were not altered by GMC preparation. During the first days of culture, CMV-T cells exhibited a lower level of CD3-induced apoptosis than did EBV-T cells. In addition, the CMV-T cells escaping initial apoptosis subsequently underwent a higher expansion rate than EBV-T cells. The differential early sensitivity to apoptosis could be in relation to the "recent activation" phenotype of EBV-T cells as evidenced by a higher level of CD69 expression. Furthermore, EBV-T cells were found to have a CD45RA-CD27+CCR7- effector memory phenotype, whereas CMV-T cells had a CD45RA+CD27-CCR7- terminal effector phenotype. Such differences could be contributive, because bulk CD8+CD27 cells had a higher expansion than did bulk CD8+CD27+ cells. Overall, ex vivo T cell culture differentially affects apoptosis, long-term proliferation, and overall survival of CMV-T and EBV-T cells. Such functional differences need to be taken into account when designing cell and/or gene therapy protocols involving ex vivo T-cell manipulation. PMID- 12730102 TI - Involvement of CXCR4 and IL-2 in the homing and retention of human NK and NK T cells to the bone marrow and spleen of NOD/SCID mice. AB - Human natural killer (NK) and NK T cells play an important role in allogeneic bone marrow (BM) transplantation and graft-versus-leukemia (GVL) effect. The mechanisms by which these cells home to the BM and spleen are not well understood. Here we show that treatment of these cells with pertussis toxin and neutralizing antibodies to the chemokine receptor CXCR4 inhibited homing of the cells to the BM, but not the spleen, of NOD/SCID mice. The retention of NK and NK T cells within the spleen and BM was dependent on Galphai signaling and CXCR4 function. The chemokine receptors CXCR4 and CXCR3 are expressed predominantly on the cell surface of NK T cells. Following activation with interleukin-2 (IL-2), the levels of CXCR4 on NK and NK T cells decreased significantly. Treatment of cells with IL-2 inhibited their migration in response to CXCL12 and their homing and retention in the BM and spleen of NOD/SCID mice. In contrast to CXCR4, the expression levels of the chemokine receptor CXCR3 and the migration of cells in response to CXCL9 and CXCL10 increased after IL-2 treatment. Thus, down regulation of CXCR4 and up-regulation of CXCR3 may direct the trafficking of cells to the site of inflammation, rather than to hematopoietic organs, and therefore may limit their alloreactive potential. PMID- 12730103 TI - 166Ho-DOTMP plus melphalan followed by peripheral blood stem cell transplantation in patients with multiple myeloma: results of two phase 1/2 trials. AB - Holmium-166 1, 4, 7, 10-tetraazcyclododecane-1, 4, 7, 10 tetramethylenephosphonate (166Ho-DOTMP) is a radiotherapeutic that localizes specifically to the skeleton and can deliver high-dose radiation to the bone and bone marrow. In patients with multiple myeloma undergoing autologous hematopoietic stem cell transplantation two phase 1/2 dose-escalation studies of high-dose 166Ho-DOTMP plus melphalan were conducted. Patients received a 30 mCi (1.110 Gbq) tracer dose of 166Ho-DOTMP to assess skeletal uptake and to calculate a patient-specific therapeutic dose to deliver a nominal radiation dose of 20, 30, or 40 Gy to the bone marrow. A total of 83 patients received a therapeutic dose of 166Ho-DOTMP followed by autologous hematopoietic stem cell transplantation 6 to 10 days later. Of the patients, 81 had rapid and sustained hematologic recovery, and 2 died from infection before day 60. No grades 3 to 4 nonhematologic toxicities were reported within the first 60 days. There were 27 patients who experienced grades 2 to 3 hemorrhagic cystitis, only 1 of whom had received continuous bladder irrigation. There were 7 patients who experienced complications considered to be caused by severe thrombotic microangiopathy (TMA). No cases of severe TMA were reported in patients receiving in 166Ho-DOMTP doses lower than 30 Gy. Approximately 30% of patients experienced grades 2 to 4 renal toxicity, usually at doses targeting more than 40 Gy to the bone marrow. Complete remission was achieved in 29 (35%) of evaluable patients. With a minimum follow up of 23 months, the median survival had not been reached and the median event free survival was 22 months. 166Ho-DOTMP is a promising therapy for patients with multiple myeloma and merits further evaluation. PMID- 12730104 TI - Repopulating defect of mismatch repair-deficient hematopoietic stem cells. AB - Mismatch repair deficiency is associated with carcinogenesis, increased spontaneous and induced mutagenesis, and resistance to methylating agents. In humans, leukemias and lymphomas arise in the background of mismatch repair deficiency, raising the possibility that hematopoiesis is abnormal as well. To address hematopoiesis in MSH2-/- mice, we collected marrow and performed serial transplantations of these cells, alone or mixed with wild-type cells, into lethally irradiated healthy mice. Transplant recipients were observed or treated with the methylating agent, temozolomide (TMZ). Methylating agent tolerance was evident by the competitive survival advantage of MSH2-/- marrow progenitors compared with wild-type cells after each TMZ exposure. However, serial repopulation by MSH2-/- cells was deficient compared with wild-type cells. In recipients of mixed populations, the MSH 2-/- cells were lost from the marrow, and mice receiving MSH2-/- cells plus TMZ could not be reconstituted in the third passage, whereas all wild-type cell recipients survived. No differences in telomere length, cell cycle distribution, or homing were observed, but an increase in microsatellite instability was seen in the MSH2-/- early progenitor colony-forming unit (CFU) and Sca+Kit+lin--derived clones. Thus, mismatch repair deficiency is associated with a hematopoietic repopulation defect and stem cell exhaustion because of accumulation of genomic instability. PMID- 12730105 TI - Decreased factor VIII levels during acetaminophen-induced murine fulminant hepatic failure. AB - During human fulminant hepatic failure (FHF) circulating levels of most hemostatic proteins fall dramatically. Concurrently, factor VIII (fVIII) procoagulant activity rises despite destruction of the hepatocytes considered responsible for fVIII synthesis. This observation suggests a role for cells other than hepatocytes in fVIII biosynthesis during FHF. We have attempted to identify nonhepatocytic sites of fVIII biosynthesis by inducing FHF in mice using acetaminophen overdose, a common cause of human FHF. Acetaminophen-treated mice consistently displayed signs characteristic of FHF, including elevated plasma aminotransferase activity. However, acetaminophen-treated mice demonstrated markedly reduced fVIII activity, contrary to the observation in human FHF. von Willebrand factor antigen levels were only mildly reduced, suggesting that the decrease in fVIII is not secondary to loss of von Willebrand factor. These results imply that there are fundamental differences in the regulation of plasma fVIII levels between humans and mice. PMID- 12730107 TI - Long targeting arms do not increase the efficiency of homologous recombination in the beta-globin locus of murine embryonic stem cells. AB - The correction of mutant beta-globin genes has long been a therapeutic goal for patients with beta-thalassemia or hemoglobinopathies. The use of homologous recombination (HR) to achieve this goal is an attractive approach because it eliminates the need to include regulatory sequences in the therapeutic construct, and it eliminates mutagenesis induced by random integration. However, HR is a very inefficient process for gene correction, and its efficiency is probably locus dependent. The length of targeting arms is thought to be a determinant of targeting efficiency, so we compared the ability of standard (8-kb) versus very long (16-, 24-, and 110-kb) regions of homology to correct a mutant murine beta globin gene in embryonic stem cells. Increasing the length of the targeting sequences did not increase the efficiency of HR in this locus, suggesting that alternative approaches will be required to improve the efficiency of this approach for globin gene correction. PMID- 12730106 TI - Comparison of molecular markers in a cohort of patients with chronic myeloproliferative disorders. AB - Decreased expression of c-MPL protein in platelets, increased expression of polycythemia rubra vera 1 (PRV-1) and nuclear factor I-B (NFIB) mRNA in granulocytes, and loss of heterozygosity on chromosome 9p (9pLOH) were described as molecular markers for myeloproliferative disorders (MPDs). To assess whether these markers are clustered in subgroups of MPDs or represent independent phenotypic variations, we simultaneously determined their status in a cohort of MPD patients. Growth of erythropoietin-independent colonies (EECs) was measured for comparison. We observed concordance between EECs and PRV-1 in MPD patients across all diagnostic subclasses, but our results indicate that EECs remain the most reliable auxiliary test for polycythemia vera (PV). In contrast, c-MPL, NFIB, and 9pLOH constitute independent variations. Interestingly, decreased c-MPL and elevated PRV-1 also were observed in patients with hereditary thrombocythemia (HT) who carry a mutation in the thrombopoietin (TPO) gene. Thus, altered c-MPL and PRV-1 expression also can arise through a molecular mechanism different from sporadic MPD. PMID- 12730108 TI - The APC-independent anticoagulant activity of protein S in plasma is decreased by elevated prothrombin levels due to the prothrombin G20210A mutation. AB - Protein S exhibits anticoagulant activity independent of activated protein C (APC). An automated factor Xa-based one-stage clotting assay was developed that enables quantification of the APC-independent activity of protein S in plasma from the ratio of clotting times (protein S ratio [pSR]) determined in the absence and presence of neutralizing antibodies against protein S. The pSR was 1.62 +/- 0.16 (mean +/- SD) in a healthy population (n = 60), independent of plasma levels of factors V, VIII, IX, and X; protein C; and antithrombin, and not affected by the presence of factor V Leiden. The pSR strongly correlates with the plasma level of protein S and is modulated by the plasma prothrombin concentration. In a group of 16 heterozygous protein S-deficient patients, the observed mean pSR (1.31 +/- 0.09) was significantly lower than the mean pSR of the healthy population, as was the pSR of plasma from carriers of the prothrombin G20210A mutation (1.47 +/- 0.21; n = 46). We propose that the decreased APC independent anticoagulant activity of protein S in plasma with elevated prothrombin levels may contribute to the thrombotic risk associated with the prothrombin G20210A mutation. PMID- 12730109 TI - Syk activation is a leukotriene B4-regulated event involved in macrophage phagocytosis of IgG-coated targets but not apoptotic cells. AB - Macrophages are called upon to ingest both IgG-coated targets and apoptotic cells. Important roles for tyrosine kinase Syk and leukotriene B4 (LTB4) are recognized in FcgammaR-mediated phagocytosis. Here we evaluated the roles of Syk and LTB4 in macrophage phagocytosis of apoptotic thymocytes versus IgG-coated erythrocytes. Macrophage ingestion of apoptotic thymocytes was not influenced by exogenous or endogenous LTB4 nor associated with Syk activation (phosphorylation). By contrast, LTB4 dose-dependently amplified FcgammaR-mediated phagocytosis as well as Syk activation. Furthermore, a role for endogenous LTB4 in Syk activation during FcgammaR-mediated phagocytosis was demonstrated using pharmacologic and genetic abrogation of 5-lipoxygenase. LTB4 was unique among 5 lipoxygenase products in this regard, since LTD4 and 5-hydroxyeicosatetraenoic acid (HETE) were unable to amplify Syk activation in response to FcgammaR engagement. Ca2+ chelation studies revealed that FcgammaR-mediated Syk activation as well as LTB4 amplification thereof was Ca2+ regulated. These 2 parallel phagocytic processes therefore exhibit initial divergence in signal transduction events, with Syk activation being an LTB4-regulated event in FcgammaR-mediated but not apoptotic cell ingestion. As LTB4 is an important proinflammatory product of macrophages, we speculate that this divergence evolved to permit FcgammaR mediated phagocytosis to proceed in an inflammatory milieu, while apoptotic cell clearance is noninflammatory. PMID- 12730110 TI - CXCL12 expression by invasive trophoblasts induces the specific migration of CD16 human natural killer cells. AB - In the maternal decidua, natural killer (NK) cells, characterized by lack of CD16, are found in direct contact with the fetal extravillous trophoblasts (EVTs). It is yet unknown which factors contribute to the specific homing of this unique NK subset to the decidua. In this study we analyze the chemokine receptor repertoire on various NK populations derived from the peripheral blood and decidua. We show that CXCR4 and CXCR3 receptors are preferentially expressed on CD16- NK subsets derived either from the peripheral blood or the decidua and that these receptors are involved in migration of all NK subsets to their ligands. We further demonstrate in vivo that invading EVTs that eventually perform endovascular invasion express CXCL12, the ligand for CXCR4, but not ligands for CXCR3. Indeed, specific accumulation of the CD16- NK cells at the expense of CD16+ cells was observed only when in vitro migration was performed with ligands for CXCR4. Finally, incubation of the peripheral blood CD16- NK cells with cytokines present in the decidua, especially interleukin 15 (IL-15), resulted in the expression of chemokine receptor repertoire similar to that observed on decidual NK cells, suggesting an additional important regulatory effect of local decidual cytokines. PMID- 12730112 TI - A defect in hematopoietic stem cell migration explains the nonrandom X-chromosome inactivation in carriers of Wiskott-Aldrich syndrome. AB - A defect in cell trafficking and chemotaxis plays an important role in the immune deficiency observed in Wiskott-Aldrich syndrome (WAS). In this report, we show that marrow cells from WAS protein (WASP)-deficient mice also have a defect in chemotaxis. Serial transplantation and competitive reconstitution experiments demonstrated that marrow cells, including hematopoietic progenitors and stem cells (HSCs), have decreased homing capacities that were associated with a defect in adhesion to collagen. During development, HSCs migrate from the liver to the marrow and the spleen, prompting us to ask if a defect in HSC homing during development may explain the skewed X-chromosome inactivation in WAS carriers. Preliminary evidence has shown that, in contrast to marrow progenitor cells, fetal liver progenitor cells from heterozygous females had a random X-chromosome inactivation. When fetal liver cells from WASP-carrier females were injected into irradiated recipients, a nonrandom inactivation of the X-chromosome was found at the level of hematopoietic progenitors and HSCs responsible for the short- and long-term hematopoietic reconstitution. Therefore, the mechanism of the skewed X chromosomal inactivation observed in WAS carriers may be related to a migration defect of WASP-deficient HSCs. PMID- 12730111 TI - Systemic regulation of Hephaestin and Ireg1 revealed in studies of genetic and nutritional iron deficiency. AB - Hephaestin is a membrane-bound multicopper ferroxidase necessary for iron egress from intestinal enterocytes into the circulation. Mice with sex-linked anemia (sla) have a mutant form of Hephaestin and a defect in intestinal basolateral iron transport, which results in iron deficiency and anemia. Ireg1 (SLC11A3, also known as Ferroportin1 or Mtp1) is the putative intestinal basolateral iron transporter. We compared iron levels and expression of genes involved in iron uptake and storage in sla mice and C57BL/6J mice fed iron-deficient, iron overload, or control diets. Both iron-deficient wild-type mice and sla mice showed increased expression of Heph and Ireg1 mRNA, compared to controls, whereas only iron-deficient wild-type mice had increased expression of the brush border transporter Dmt1. Unlike iron-deficient mice, sla mouse enterocytes accumulated nonheme iron and ferritin. These results indicate that Dmt1 can be modulated by the enterocyte iron level, whereas Hephaestin and Ireg1 expression respond to systemic rather than local signals of iron status. Thus, the basolateral transport step appears to be the primary site at which the small intestine responds to alterations in body iron requirements. PMID- 12730113 TI - Sirolimus, tacrolimus, and low-dose methotrexate for graft-versus-host disease prophylaxis in mismatched related donor or unrelated donor transplantation. AB - We studied the feasibility and activity of adding sirolimus to tacrolimus and low dose methotrexate as graft-versus-host disease (GVHD) prophylaxis in recipients of alternative donor transplants. Forty-one patients with hematologic malignancies were conditioned with cyclophosphamide and total body irradiation. Marrow stem cells were from an HLA-A, -B, and -DR compatible, unrelated donor (n = 26, 68%), from a 5 of 6 antigen-matched unrelated donor (n = 8, 20%), or from a 5 of 6 antigen-matched family member (n = 5, 12%). Therapeutic serum levels of sirolimus were attained in most patients. All evaluable patients engrafted. An absolute neutrophil count of 500/microL was achieved on day +18 (range, 11-32 days). Sustained platelet counts of more than 20 000/ microL were attained on day +29 (range, 14-98 days). Grades 0-I acute GVHD occurred in 75% of patients. Grades II, III, and IV acute GVHD occurred in 13%, 8%, and 5%, respectively (total grades II-IV GVHD, 26%). Median survival is 366 days (95% CI 185, not estimable) and actuarial survival at 1 year is 52%. Oral sirolimus is tolerable, adequate blood levels are achievable, and there is a low rate of acute GVHD compared with historical data in this high-risk population. This novel agent is worthy of further study in allogeneic transplantation. PMID- 12730114 TI - Molecular analyses of patients with hyperferritinemia and normal serum iron values reveal both L ferritin IRE and 3 new ferroportin (slc11A3) mutations. AB - Unexplained hyperferritinemia is a common clinical finding, even in asymptomatic persons. When early onset bilateral cataracts are also present, the hereditary hyperferritinemia-cataract syndrome (HHCS), because of heterozygous point mutation in the L ferritin iron-responsive element (IRE) sequence, can be suspected. We sequenced the L ferritin exon 1 in 52 DNA samples from patients referred to us for molecular diagnosis of HHCS. We identified 24 samples with a point mutation/deletion in the IRE. For the 28 samples in which no IRE mutation was present, we also genotyped HFE mutations and sequenced both H ferritin and ferroportin genes. We found an increased frequency of His63Asp heterozygotes (12 of 28) but no H ferritin mutations. We identified 3 new ferroportin mutations, producing, respectively, Asp157Gly, Gln182His, and Gly323Val amino acid replacements, suggesting that these patients have dominant type 4 hemochromatosis. This study demonstrates that both L ferritin IRE and ferroportin mutations can account for isolated hyperferritinemia. The presence of cataract does not permit the unambiguous identification of patients with HHCS, although the existence of a family history of cataract was only encountered in these patients. This raises the intriguing possibility that lens ferritin accumulation might be a factor contributing to age-related cataract in the general population. Additional causes of isolated hyperferritinemia remain to be identified. PMID- 12730115 TI - Classification of pediatric acute lymphoblastic leukemia by gene expression profiling. AB - Contemporary treatment of pediatric acute lymphoblastic leukemia (ALL) requires the assignment of patients to specific risk groups. We have recently demonstrated that expression profiling of leukemic blasts can accurately identify the known prognostic subtypes of ALL, including T-cell lineage ALL (T-ALL), E2A-PBX1, TEL AML1, MLL rearrangements, BCR-ABL, and hyperdiploid karyotypes with more than 50 chromosomes. As the next step toward developing this methodology into a frontline diagnostic tool, we have now analyzed leukemic blasts from 132 diagnostic samples using higher density oligonucleotide arrays that allow the interrogation of most of the identified genes in the human genome. Nearly 60% of the newly identified subtype discriminating genes are novel markers not identified in our previous study, and thus should provide new insights into the altered biology underlying these leukemias. Moreover, a proportion of the newly selected genes are highly ranked as class discriminators, and when incorporated into class-predicting algorithms resulted in an overall diagnostic accuracy of 97%. The performance of an array containing the identified discriminating genes should now be assessed in frontline clinical trials in order to determine the accuracy, practicality, and cost effectiveness of this methodology in the clinical setting. PMID- 12730116 TI - Increased apoptosis in bone marrow B lymphocytes but not T lymphocytes in myelodysplastic syndrome. AB - The hallmark of myelodysplastic syndrome (MDS) is enhanced apoptosis in myeloid, erythroid, and megakaryocytic cells in the bone marrow leading to ineffective hematopoiesis. Recent studies suggested that immunological and microenvironmental factors play a role in the pathophysiology of this disease. We report a significant increase in apoptosis in bone marrow B lymphocytes in MDS as compared to that found in acute myeloid leukemia and healthy controls. Furthermore, we demonstrate that patients with refractory anemia with excess blasts in transformation (RAEB-T) had apoptosis levels in lymphocytes similar to those seen in other subtypes of MDS. Our findings suggest that the alterations in B lymphocytes in the form of increased apoptosis can be seen in MDS and support the concept that immune modulation plays a role in the pathophysiology of MDS. PMID- 12730117 TI - Double jeopardy from a single translocation: deletions of the derivative chromosome 9 in chronic myeloid leukemia. AB - Chronic myeloid leukemia (CML) is characterized by formation of a BCR-ABL fusion gene, usually as a consequence of the Philadelphia (Ph) translocation between chromosomes 9 and 22. Recently the development of new fluorescence in-situ hybridization (FISH) techniques has allowed identification of unexpected deletions of the reciprocal translocation product, the derivative chromosome 9, in 10% to 15% of patients with CML. These deletions are large, span the translocation breakpoint, and occur at the same time as the Ph translocation. Such deletions therefore give rise to previously unsuspected molecular heterogeneity from the very beginning of this disease, and there is mounting evidence for similar deletions associated with other translocations. Several studies have demonstrated that CML patients who carry derivative chromosome 9 deletions exhibit a more rapid progression to blast crisis and a shorter survival. Deletion status is independent of, and more powerful than, the Sokal and Hasford/European prognostic scoring systems. The poor prognosis associated with deletions is seen in patients treated with hydroxyurea or interferon, and preliminary evidence suggests that patients with deletions may also have a worse outcome than nondeleted patients following stem cell transplantation or treatment with imatinib. Poor outcome cannot be attributed to loss of the reciprocal ABL BCR fusion gene expression alone, and is likely to reflect loss of one or more critical genes within the deleted region. The molecular heterogeneity associated with the Philadelphia translocation provides a new paradigm with potential relevance to all malignancies associated with reciprocal chromosomal translocations and/or fusion gene formation. PMID- 12730118 TI - Murine GPVI stimulates weak integrin activation in PLCgamma2-/- platelets: involvement of PLCgamma1 and PI3-kinase. AB - Collagen stimulates platelet activation through a tyrosine kinase-based pathway downstream of the glycoprotein VI (GPVI)-Fc receptor (FcR) gamma-chain complex. Genetic ablation of FcR gamma-chain results in a complete inhibition of aggregation to collagen. In contrast, a steady increase in light transmission is induced by collagen in phospholipase Cgamma2-deficient (PLCgamma2-/-) platelets in a Born aggregometer, indicating a weak level of activation. This increase is inhibited partially in the presence of an alpha2beta1-blocking antibody or an alphaIIbbeta3 antagonist and completely by a combination of the 2 inhibitors. It is also abolished by the Src kinase inhibitor PP1 and reduced in the presence of the phosphatidylinositol (PI) 3-kinase inhibitor wortmannin. The GPVI-specific agonists convulxin and collagen-related peptide (CRP) also stimulate weak aggregation in PLCgamma2-/- platelets, which is inhibited by wortmannin and PP1. Collagen and CRP stimulate tyrosine phosphorylation of PLCgamma1 at its regulatory site, Tyr 783, in murine but not in human platelets through a Src kinase-dependent pathway. Adhesion of PLCgamma2-/- platelets to a collagen monolayer is severely reduced at a shear rate of 800 s-1, relative to controls, whereas it is abolished in FcR gamma-chain-/- platelets. These results provide strong evidence that engagement of GPVI stimulates limited integrin activation in PLCgamma2-/- platelets via PLCgamma1 and PI3-kinase. PMID- 12730119 TI - The nonenzymatic subunit of pseutarin C, a prothrombin activator from eastern brown snake (Pseudonaja textilis) venom, shows structural similarity to mammalian coagulation factor V. AB - Pseutarin C is a group C prothrombin activator from the venom of the eastern brown snake Pseudonaja textilis. It is a multi-subunit protein complex consisting of catalytic and nonenzymatic subunits similar to coagulation factor Xa and factor Va, respectively. Here we describe the complete sequence of the nonenzymatic subunit. Based on the partial amino acid sequence of the nonenzymatic subunit, degenerate primers were designed. Using a "walking" strategy based on sequentially designed primers, we determined the complete cDNA sequence of the nonenzymatic subunit. The cDNA encodes a protein of 1461 amino acid residues, which includes a 30-residue signal peptide, a mature protein of 1430 amino acid residues, and a stop codon. cDNA blot analysis showed a single transcript of approximately 4.6 kb. The deduced amino acid sequence shows approximately 50% identity to mammalian factor V and by homology has a similar domain structure consisting of domains A1-A2-B-A3-C1-C2. Interestingly, the B domain of pseutarin C is shorter than that of mammalian factor V (FV). Although most of the proteolytic activation sites are conserved, 2 of 3 proteolytic sites cleaved by activated protein C are mutated, and thus activated protein C is not able to inactivate this procoagulant toxin. The predicted posttranslational modifications, including disulfide bonds, N-glycosylation, phosphorylation, and sulfation, in pseutarin C are significantly different compared with bovine factor V. Thus, our data demonstrate that the nonenzymatic subunit of group C prothrombin activators is structurally similar to mammalian FV. PMID- 12730120 TI - Cyclin D1 overexpression is a favorable prognostic variable for newly diagnosed multiple myeloma patients treated with high-dose chemotherapy and single or double autologous transplantation. AB - We used a sensitive real-time reverse transcription-polymerase chain reaction assay to quantify cyclin D1 mRNA levels in bone marrow samples collected at diagnosis from 74 newly diagnosed multiple myeloma (MM) patients who were randomized to undergo either single or double autologous peripheral blood stem cell transplantation as part of first-line therapy for their malignancy. In 46 cases, fluorescence in situ hybridization (FISH) analysis and/or conventional cytogenetics were performed to detect chromosome 11 abnormalities. Patients with the t(11;14) or trisomy 11 significantly overexpressed cyclin D1 (P <.0001) in comparison with patients without 11q abnormalities, who had cyclin D1 mRNA levels similar to healthy donors. Overall, 32 (43%) of 74 patients showed cyclin D1 overexpression. No difference was found between cyclin D1-positive (group A) and cyclin D1-negative (group B) patients with respect to presenting clinical and laboratory characteristics, including chromosome 13 abnormalities, as well as to response to therapy and overall survival, both of which were calculated on an intent-to-treat basis. Patients who overexpressed cyclin D1 had significantly longer duration of remission in comparison with patients who did not (41 vs 26 months, respectively; P =.02). As a result, median event-free survival (EFS) was longer in group A than in group B (33 vs 24 months, respectively; P =.055). We concluded that cyclin D1 overexpression is closely associated with 11q abnormalities and identifies a subset of MM patients who are more likely to have prolonged duration of remission and EFS following autologous transplantation. PMID- 12730121 TI - Altered localization of Drosophila Smoothened protein activates Hedgehog signal transduction. AB - Hedgehog (Hh) signaling is critical for many developmental events and must be restrained to prevent cancer. A transmembrane protein, Smoothened (Smo), is necessary to transcriptionally activate Hh target genes. Smo activity is blocked by the Hh transmembrane receptor Patched (Ptc). The reception of a Hh signal overcomes Ptc inhibition of Smo, activating transcription of target genes. Using Drosophila salivary gland cells in vivo and in vitro as a new assay for Hh signal transduction, we investigated the regulation of Hh-triggered Smo stabilization and relocalization. Hh causes Smo to move from internal membranes to the cell surface. Relocalization is protein synthesis-independent and occurs within 30 min of Hh treatment. Ptc and the kinesin-related protein Costal2 (Cos2) cause internalization of Smo, a process that is dependent on both actin and microtubules. Disruption of endocytosis by dominant negative dynamin or Rab5 prevents Smo internalization. Fly versions of Smo mutants associated with human tumors are constitutively present at the cell surface. Forced localization of Smo at the plasma membrane activates Hh target gene transcription. Conversely, trapping of activated Smo mutants in the ER prevents Hh target gene activation. Control of Smo localization appears to be a crucial step in Hh signaling in Drosophila. PMID- 12730122 TI - A complex of LIN-5 and GPR proteins regulates G protein signaling and spindle function in C elegans. AB - The Caenorhabditis elegans coiled-coil protein LIN-5 mediates several processes in cell division that depend on spindle forces, including alignment and segregation of chromosomes and positioning of the spindle. Here, we describe two closely related proteins, GPR-1 and GPR-2 (G protein regulator), which associate with LIN-5 in vivo and in vitro and depend on LIN-5 for localization to the spindle and cell cortex. GPR-1/GPR-2 contain a GoLoco/GPR motif that mediates interaction with GDP-bound Galpha(i/o). Inactivation of lin-5, gpr-1/gpr-2, or the Galpha(i/o) genes goa-1 and gpa-16 all cause highly similar chromosome segregation and spindle positioning defects, indicating a positive role for the LIN-5 and GPR proteins in G protein signaling. The lin-5 and gpr-1/gpr-2 genes appear to act downstream of the par polarity genes in the one- and two-cell stages and downstream of the tyrosine kinase-related genes mes-1 and src-1 at the four-cell stage. Together, these results indicate that GPR-1/GPR-2 in association with LIN-5 activate G protein signaling to affect spindle force. Polarity determinants may regulate LIN-5/GPR/Galpha locally to create the asymmetric forces that drive spindle movement. Results in C. elegans and other species are consistent with a novel model for receptor-independent activation of Galpha(i/o) signaling. PMID- 12730123 TI - Notch activity induces Nodal expression and mediates the establishment of left right asymmetry in vertebrate embryos. AB - Left-sided expression of Nodal in the lateral plate mesoderm is a conserved feature necessary for the establishment of normal left-right asymmetry during vertebrate embryogenesis. By using gain- and loss-of-function experiments in zebrafish and mouse, we show that the activity of the Notch pathway is necessary and sufficient for Nodal expression around the node, and for proper left-right determination. We identify Notch-responsive elements in the Nodal promoter, and unveil a direct relationship between Notch activity and Nodal expression around the node. Our findings provide evidence for a mechanism involving Notch activity that translates an initial symmetry-breaking event into asymmetric gene expression. PMID- 12730125 TI - Endothelial dysfunction and LOX-1: forty years from muscle to endothelium. PMID- 12730124 TI - Notch signaling regulates left-right asymmetry determination by inducing Nodal expression. AB - Generation of left-right asymmetry is an integral part of the establishment of the vertebrate body plan. Here we show that the Notch signaling pathway plays a primary role in the establishment of left-right asymmetry in mice by directly regulating expression of the Nodal gene. Embryos mutant for the Notch ligand Dll1 or doubly mutant for the Notch1 and Notch2 receptors exhibit multiple defects in left-right asymmetry. Analysis of the enhancer regulating node-specific Nodal expression revealed the presence of binding sites for the RBP-J protein, the primary transcriptional mediator of Notch signaling. Mutation of these sites destroyed the ability of this enhancer to direct node-specific gene expression in transgenic mice. Our results demonstrate that Dll1-mediated Notch signaling is essential for generation of left-right asymmetry, and that the Notch pathway acts upstream of Nodal expression during left-right asymmetry determination in mice. PMID- 12730126 TI - Surviving hypoxia: the importance of rafts, anchors, and fluidity. PMID- 12730127 TI - Decisions, decisions...SRF coactivators and smooth muscle myogenesis. PMID- 12730128 TI - Matrix metalloproteinases and tissue inhibitors of metalloproteinases: structure, function, and biochemistry. AB - Matrix metalloproteinases (MMPs), also designated matrixins, hydrolyze components of the extracellular matrix. These proteinases play a central role in many biological processes, such as embryogenesis, normal tissue remodeling, wound healing, and angiogenesis, and in diseases such as atheroma, arthritis, cancer, and tissue ulceration. Currently 23 MMP genes have been identified in humans, and most are multidomain proteins. This review describes the members of the matrixin family and discusses substrate specificity, domain structure and function, the activation of proMMPs, the regulation of matrixin activity by tissue inhibitors of metalloproteinases, and their pathophysiological implication. PMID- 12730129 TI - Genetics of human laterality disorders: insights from vertebrate model systems. AB - Many internal organs in the vertebrate body are asymmetrically oriented along the left-right (L-R) body axis. Organ asymmetry and some components of the molecular signaling pathways that direct L-R development are highly conserved among vertebrate species. Although individuals with full reversal of organ L-R asymmetry (situs inversus totalis) are healthy, significant morbidity and mortality is associated with perturbations in laterality that result in discordant orientation of organ systems and complex congenital heart defects. In humans and other vertebrates, genetic alterations of L-R signaling pathways can result in a wide spectrum of laterality defects. In this review we categorize laterality defects in humans, mice, and zebrafish into specific classes based on altered patterns of asymmetric gene expression, organ situs defects, and midline phenotypes. We suggest that this classification system provides a conceptual framework to help consolidate the disparate laterality phenotypes reported in humans and vertebrate model organisms, thereby refining our understanding of the genetics of L-R development. This approach helps suggest candidate genes and genetic pathways that might be perturbed in human laterality disorders and improves diagnostic criteria. PMID- 12730130 TI - Reconstructing MYC. PMID- 12730131 TI - A role for a novel 'trans-pseudoknot' RNA-RNA interaction in the functional dimerization of human telomerase. AB - The integral RNA (hTER) of the human telomerase ribonucleoprotein has a conserved secondary structure that contains a potential pseudoknot. Here we examine the role of an intermolecular hTER-hTER interaction in the previously reported functional dimerization of telomerase. We provide evidence that the two conserved, complementary sequences of one stem of the hTER pseudoknot domain can pair intermolecularly in vitro, and that formation of this stem as part of a novel "trans-pseudoknot" is required for telomerase to be active in its dimeric form. Such RNA-RNA interaction mirrors a known property of retroviral reverse transcriptases, which use homodimeric viral genomic RNA substrates. PMID- 12730132 TI - Latent ClpX-recognition signals ensure LexA destruction after DNA damage. AB - The DNA-damage response genes in bacteria are up-regulated when LexA repressor undergoes autocatalytic cleavage stimulated by activated RecA protein. Intact LexA is stable to intracellular degradation but its auto-cleavage fragments are degraded rapidly. Here, both fragments of LexA are shown to be substrates for the ClpXP protease. ClpXP recognizes these fragments using sequence motifs that flank the auto-cleavage site but are dormant in intact LexA. Furthermore, ClpXP degradation of the LexA-DNA-binding fragment is important to cell survival after DNA damage. These results demonstrate how one protein-processing event can activate latent protease recognition signals, triggering a cascade of protein turnover in response to environmental stress. PMID- 12730134 TI - GINS, a novel multiprotein complex required for chromosomal DNA replication in budding yeast. AB - Eukaryotic chromosomal DNA replication requires a two-step assembly of replication proteins on origins; formation of the prereplicative complex (pre-RC) in late M and G1 phases of the cell cycle, and assembly of other replication proteins in S phase to load DNA polymerases to initiate DNA synthesis. In budding yeast, assembly of Dpb11 and the Sld3-Cdc45 complex on the pre-RC at origins is required for loading DNA polymerases. Here we describe a novel replication complex, GINS (Go, Ichi, Nii, and San; five, one, two, and three in Japanese), in budding yeast, consisting of Sld5, Psf1 (partner of Sld five 1), Psf2, and Psf3 proteins, all of which are highly conserved in eukaryotic cells. Since the conditional mutations of Sld5 and Psf1 confer defect of DNA replication under nonpermissive conditions, GINS is suggested to function for chromosomal DNA replication. Consistently, in S phase, GINS associates first with replication origins and then with neighboring sequences. Without GINS, neither Dpb11 nor Cdc45 associates properly with chromatin DNA. Conversely, without Dpb11 or Sld3, GINS does not associate with origins. Moreover, genetic and two-hybrid interactions suggest that GINS interacts with Sld3 and Dpb11. Therefore, Dpb11, Sld3, Cdc45, and GINS assemble in a mutually dependent manner on replication origins to initiate DNA synthesis. PMID- 12730133 TI - A novel ring-like complex of Xenopus proteins essential for the initiation of DNA replication. AB - We have identified Xenopus homologs of the budding yeast Sld5 and its three interacting proteins. These form a novel complex essential for the initiation of DNA replication in Xenopus egg extracts. The complex binds to chromatin in a manner dependent on replication licensing and S-phase CDK. The chromatin binding of the complex and that of Cdc45 are mutually dependent and both bindings require Xenopus Cut5, the yeast homolog of which interacts with Sld5. On replicating chromatin the complex interacts with Cdc45 and MCM, putative components of replication machinery. Electron microscopy further reveals that the complex has a ring-like structure. These results suggest that the complex plays an essential role in the elongation stage of DNA replication as well as the initiation stage. PMID- 12730135 TI - The master regulator for entry into sporulation in Bacillus subtilis becomes a cell-specific transcription factor after asymmetric division. AB - Gene transcription at the onset of sporulation in Bacillus subtilis is governed by Spo0A, a member of the response regulator family of transcription factors. Spo0A is traditionally viewed as the master regulator for entry into development. We now report that Spo0A continues to function after the initiation phase of sporulation and that it becomes a cell-specific transcription factor when the sporangium is divided into a mother cell and forespore. We observed that (1) Spo0A and Spo0A-directed gene transcription reached high levels in the mother cell; (2) an activated form of Spo0A impaired sporulation when produced in the forespore but not when produced in the mother cell; and (3) an inhibitor of Spo0A called Spo0A-N impaired sporulation and Spo0A-directed transcription when produced in the mother cell but not when produced in the forespore. Spo0A-N, which corresponds to the NH(2)-terminal domain of Spo0A, was shown to compete with the full-length response regulator for phosphorylation by the phosphorelay protein Spo0B. We propose that Spo0A is the earliest-acting transcription factor in the mother-cell line of gene expression and that in terms of abundance and transcriptional activity Spo0A may function predominantly as a cell-specific regulatory protein. PMID- 12730137 TI - Inflated impacts of medication use technology assumed in simulating reduced adverse drug events. PMID- 12730136 TI - Molecular analysis of the LATERAL SUPPRESSOR gene in Arabidopsis reveals a conserved control mechanism for axillary meristem formation. AB - In seed plants, shoot branching is initiated by the formation of new meristems in the axils of leaves, which subsequently develop into new axes of growth. This study describes the genetic control of axillary meristem formation by the LATERAL SUPPRESSOR (LAS) gene in Arabidopsis thaliana. las mutants show a novel phenotype that is characterized by the inability to form lateral shoots during vegetative development. The analysis shows that axillary meristem formation is differently regulated during different phases of development. During reproductive development, axillary meristems initiate in close proximity to the shoot apical meristem and do not require LAS function. In contrast, during the vegetative phase, axillary meristems initiate at a distance to the SAM and require LAS function. This control mechanism is conserved between the distantly related species tomato and Arabidopsis. Monitoring the patterns of LAS and SHOOT MERISTEMLESS transcript accumulation allowed us to identify early steps in the development of leaf axil identity, which seem to be a prerequisite for axillary meristem initiation. Other regulators of shoot branching, like REVOLUTA and AUXIN RESISTANT 1, act downstream of LAS. The results are discussed in the context of the "detached meristem" and the "de novo formation" concepts of axillary meristem formation. PMID- 12730139 TI - Pharmacokinetic and pharmacodynamic profile of linezolid in healthy volunteers and patients with Gram-positive infections. AB - The pharmacokinetics and pharmacodynamics of linezolid have been extensively investigated in laboratory models, healthy volunteers and patients. Three formulations exist: an intravenous (iv) form, film-coated tablets and an oral suspension. Linezolid can be assayed in serum and body fluids by HPLC and has good bioavailability with a Cmax at 0.5-2 h. The protein binding is 31%, and the volume of distribution is 30-50 L with adequate to good tissue penetration into skin blister fluids, bone, muscle, fat, alveolar cells, lung extracellular lining fluid and CSF. There are two major metabolites of linezolid (PNU-142586 and PNU 142300). Non-enzymic formation of PNU-142586 is the rate-limiting step in the clearance of linezolid, and linezolid and its two main metabolites plus several minor ones are all excreted in the urine. Dose linearity is evident in the Cmax and AUC across a wide range of doses. Gender and age have little effect on pharmacokinetics, but children have greater plasma clearance and volume of distribution and hence, have lower serum concentrations for equivalent doses in adults. No dose modification is needed in mild to moderate liver disease or any degree of renal impairment; however, both PNU-142586 and PNU-142300 accumulate in renal failure. Linezolid is bacteriostatic with a significant post-antibiotic effect against the key pathogens. In animal models of infection, the time the antibiotic concentration exceeds the MIC (t > MIC) helps to determine outcome, and a t > MIC of 40% is predictive of a bacteriostatic effect for both staphylococci and pneumococci. In man, t > MIC and AUC/MIC have been related to bacteriological and clinical outcomes. AUC and length of treatment are also related to the risk of thrombocytopenia. PMID- 12730138 TI - Linezolid in vitro: mechanism and antibacterial spectrum. AB - Oxazolidinones are prominent among the new Gram-positive antimicrobial agents now becoming available. They were discovered by DuPont Pharmaceuticals in the late 1980s but linezolid, the first analogue suitable for development, was found only when the family was re-examined by Pharmacia in the 1990s. Oxazolidinones bind to the 50S subunit of the prokaryotic ribosome, preventing formation of the initiation complex for protein synthesis. This is a novel mode of action; other protein synthesis inhibitors either block polypeptide extension or cause misreading of mRNA. Linezolid MICs vary slightly with the test method, laboratory, and significance attributed to thin hazes of bacterial survival, but all workers find that the susceptibility distributions are narrow and unimodal, with MIC values between 0.5 and 4 mg/L for streptococci, enterococci and staphylococci. Full activity is retained against Gram-positive cocci resistant to other antibiotics, including methicillin-resistant staphylococci and vancomycin resistant enterococci. MICs are 4-8 mg/L for Moraxella, Pasteurella and Bacteroides spp. but other Gram-negative bacteria are resistant as a result of endogenous efflux activity. Resistance is difficult to select in vitro but has been reported during therapy in a few enterococcal infections and in two MRSA cases to date; the mechanism entails mutation of the 23S rRNA that forms the binding site for linezolid. Risk factors for selection of resistance include indwelling devices, undrained foci, protracted therapy and underdosage. PMID- 12730140 TI - Efficacy of linezolid versus comparator therapies in Gram-positive infections. AB - Treatment of Gram-positive bacterial infections is currently a therapeutic challenge because many of these pathogens are now resistant to standard antimicrobial agents. The emergence of multidrug-resistant, Gram-positive pathogens emphasizes the need for new antimicrobial therapy. Linezolid is an oxazolidinone antibiotic with a novel mechanism of action that works by inhibiting bacterial protein synthesis by blocking formation of the initiation complex. It is active against Gram-positive organisms resistant to other antibiotics, including methicillin-resistant Staphylococcus aureus (MRSA), penicillin-resistant Streptococcus pneumoniae and vancomycin-resistant enterococci (VRE). Results are encouraging from several large-scale, randomized, Phase III trials comparing the efficacy and safety of linezolid with standard comparator agents for the treatment of nosocomial pneumonia, community-acquired pneumonia, skin and skin structure infections, and infections due to MRSA and VRE. Intravenous/oral linezolid is a promising antimicrobial agent and provides the clinician with an additional treatment option, particularly among the limited therapies for resistant Gram-positive bacterial infections. PMID- 12730141 TI - Impact of methicillin-resistant Staphylococcus aureus infections on key health economic outcomes: does reducing the length of hospital stay matter? AB - The clinical and economic impact of methicillin-resistant staphylococcal infections on the patient, the hospital and the community is significant and continues to increase. Methicillin-resistant Staphylococcus aureus infections, in particular, represent a substantial burden of resistant infections in the hospital. As such, it is important to analyse the cost parameters associated with an episode of infection. Key determinants of the total cost of an episode of infection include fixed costs and hotel costs; the contribution of antimicrobial therapy, including drug acquisition and delivery costs, is comparatively marginal. Therefore, decreasing the hospital length of stay by promoting earlier hospital discharge will significantly reduce overall costs and, in effect, increase the efficiency and cost-effectiveness of the hospital. Linezolid, the first marketed agent of a new class of oxazolidinone antibiotics, is effective in the treatment of serious Gram-positive infections. Its availability in both intravenous (iv) and oral formulations (the latter of which is 100% bioavailable) facilitates early discharge from hospital. Hospitalized patients with methicillin resistant staphylococcal infections who were treated with either linezolid or vancomycin demonstrated no significant differences in clinical outcome or mortality. However, compared with vancomycin, administration of linezolid reduced the duration of iv therapy and increased the chance of being discharged during the first week of hospitalization. These effects were most pronounced in the subset of patients with skin and soft tissue infections. Knowledge of these data may influence prescribing practices at the individual and organizational levels and, in turn, reduce the economic burden associated with MRSS infections. PMID- 12730142 TI - Safety and tolerability of linezolid. AB - Clinical trials have shown that linezolid (600 mg twice daily in adults) is safe and generally well tolerated for up to 28 days. Drug-related adverse events, which are typically mild to moderate in intensity and of limited duration, include diarrhoea, nausea and headache in adults, and diarrhoea, loose stools and vomiting in children. Clostridium difficile-related complications with linezolid are uncommon. Linezolid is a weak, reversible monoamine oxidase inhibitor: foods containing high concentrations of tyramine should be avoided, and linezolid should be used with caution in patients taking adrenergic or serotonergic agents or in those with uncontrolled hypertension. In the majority of patients, linezolid has minimal adverse effects on blood chemistry or haematology. There have been case reports of reversible thrombocytopenia, anaemia and neutropenia associated with linezolid therapy. In Phase III studies, 2.4% of patients treated with linezolid and 1.5% of patients treated with comparator drugs developed reversible thrombocytopenia (P = 0.066), but there was no evidence of an increased risk of agranulocytosis, aplastic anaemia or other irreversible blood dyscrasias. Reduced platelet counts were associated with linezolid treatment for >/=2 weeks; complete blood counts should be monitored weekly in patients receiving linezolid for more than 14 days and treatment should be discontinued if there is evidence of myelosuppression. PMID- 12730143 TI - M1/70 attenuates blood-borne neutrophil oxidants, activation, and myofiber damage following stretch injury. AB - The purpose of this study was to determine the role of the CD11b-dependent respiratory burst in neutrophil oxidant generation and activation, interleukin-8 (IL-8) production, and myofiber damage after muscle stretch injury by using the monoclonal antibody M1/70 to block this pathway. Twelve male New Zealand White rabbits were randomly assigned to a treatment group: M1/70 (n = 6), IgG isotype control (n = 3), or saline control (n = 3). After intravenous injection of the assigned agent under gas anesthesia, a standardized single-stretch injury was created in the right tibialis anterior, whereas the left tibialis anterior underwent a sham surgery. Blood-borne neutrophil oxidant generation and CD11b receptor density and plasma IL-8 levels were measured pre- and 24 h postinjury. Damage was assessed histologically at the hematoma site by counting torn myofibers. M1/70 group demonstrated decreased blood-borne neutrophil oxidant generation (P < 0.05) and CD11b receptor density (P < 0.05), an increase in plasma IL-8 concentration (P < 0.01), and less torn myofibers (P < 0.01) compared with IgG isotype or saline control groups. These data indicate that 1). CD11b dependent respiratory burst is a major source of oxidants produced by the neutrophil, and that treatment with M1/70 2). attenuates neutrophil activation status, 3). increases plasma IL-8 concentration, and 4). minimizes myofiber damage 24 h postmuscle stretch injury. PMID- 12730144 TI - Modulation of cholinergic responsiveness through the [beta]-adrenoceptor signal transmission pathway in bovine trachealis. AB - The effects of pharmacological stimulation at different levels of the beta adrenoceptor (AR) pathway, including the receptor, the receptor-coupled Gs protein, and adenylyl cyclase, were studied by simultaneous measurements of acetylcholine (ACh) release and isometric force evoked by electric stimulation in isolated bovine trachealis. The beta-AR agonists isoproterenol (10-6 and 10-5 M) and salbutamol (10-7 to 10-5 M) significantly attenuated both ACh release and contractile force. Forskolin, at 10-6 M, significantly increased ACh release without effect on contractile force, whereas at 10-5 M it increased ACh release but significantly decreased force. Activation of Gs protein by cholera toxin (10 microg/ml) significantly attenuated both ACh release and contractile force, but its effect on ACh release was abolished by calcium-activated potassium (KCa) channel blocker iberiotoxin (10-7 M). The KCa-channel opener NS-1619 (10-4 M) attenuated significantly both ACh release and contractile force. It is concluded that beta-AR agonists attenuate cholinergic neurotransmission in isolated bovine trachealis model by a mechanism not involving cAMP but KCa channels. PMID- 12730145 TI - Finger dexterity, skin temperature, and blood flow during auxiliary heating in the cold. AB - The primary purpose of the present study was to compare the effectiveness of two forms of hand heating and to discuss specific trends that relate finger dexterity performance to variables such as finger skin temperature (T(fing)), finger blood flow (Q(fing)), forearm skin temperature (T(fsk)), forearm muscle temperature (Tfmus), mean weighted body skin temperature (Tsk), and change in body heat content (DeltaH(b)). These variables along with rate of body heat storage, toe skin temperature, and change in rectal temperature were measured during direct and indirect hand heating. Direct hand heating involved the use of electrically heated gloves to keep the fingers warm (heated gloves condition), whereas indirect hand heating involved warming the fingers indirectly by actively heating the torso with an electrically heated vest (heated vest condition). Seven men (age 35.6 +/- 5.6 yr) were subjected to each method of hand heating while they sat in a chair for 3 h during exposure to -25 degrees C air. Q(fing) was significantly (P < 0.05) higher during the heated vest condition compared with the heated gloves condition (234 +/- 28 and 33 +/- 4 perfusion units, respectively), despite a similar T(fing) (which ranged between 28 and 35 degrees C during the 3-h exposure). Despite the difference in Q(fing), there was no significant difference in finger dexterity performance. Therefore, finger dexterity can be maintained with direct hand heating despite a low Q(fing). DeltaH(b), Tsk, and T(fmus) reached a low of -472 +/- 18 kJ, 28.5 +/- 0.3 degrees C, and 29.8 +/- 0.5 degrees C, respectively, during the heated gloves condition, but the values were not low enough to affect finger dexterity. PMID- 12730146 TI - Substance P released from intrinsic airway neurons contributes to ozone-enhanced airway hyperresponsiveness in ferret trachea. AB - Exposure to ozone (O3) induces airway hyperresponsiveness mediated partly through the release of substance P (SP) from nerve terminals in the airway wall. Although substantial evidence suggests that SP is released by sensory nerves, SP is also present in neurons of airway ganglia. The purpose of this study was to investigate the role of intrinsic airway neurons in O3-enhanced airway responsiveness in ferret trachea. To remove the effects of sensory innervation, segments of ferret trachea were maintained in culture conditions for 24 h before in vitro exposure to 2 parts/million of O3 or air for 1 h. Sensory nerve depletion was confirmed by showing that capsaicin did not affect tracheal smooth muscle responsiveness to cholinergic agonist or contractility responses to electrical field stimulation (EFS). Contractions of isolated tracheal smooth muscle to EFS were significantly increased after in vitro O3 exposure, but the constrictor response to cholinergic agonist was not altered. Pretreatment with CP 99994, an antagonist of the neurokinin 1 receptor, attenuated the increased contraction to EFS after O3 exposure but had no effect in the air exposure group. The number of SP-positive neurons in longitudinal trunk ganglia, the extent of SP innervation to superficial muscular plexus nerve cell bodies, and SP nerve fiber density in tracheal smooth muscle all increased significantly after O3 exposure. The results show that release of SP from intrinsic airway neurons contributes to O3-enhanced tracheal smooth muscle responsiveness by facilitating acetylcholine release from cholinergic nerve terminals. PMID- 12730147 TI - TNF-[alpha] modulates murine tracheal rings responsiveness to G-protein-coupled receptor agonists and KCl. AB - Although the mechanisms that underlie airway hyperresponsiveness in asthma are complex and involve a variety of factors, evidence now suggests that intrinsic abnormalities in airway smooth muscle (ASM) may play an important role. We previously reported that TNF-alpha, a cytokine involved in asthma, augments G protein-coupled receptor (GPCR) agonist-evoked calcium responses in cultured ASM cells. Here we have extended our previous studies by investigating whether TNF alpha also modulates the contractile and relaxant responses to GPCR activation using cultured murine tracheal rings. We found that in tracheal rings treated with 50 ng/ml TNF-alpha, carbachol-induced isometric force was significantly increased by 30% compared with those treated with diluent alone (P < 0.05). TNF alpha also augmented KCl-induced force generation by 70% compared with rings treated with diluent alone (P < 0.01). The enhancing effect of TNF-alpha on carbachol-induced isometric force generation was completely abrogated in the tracheal rings obtained from TNF-alpha receptor (TNFR)1-deficient mice and in control rings treated with a TNF-alpha mutant that solely activates TNFR2. TNF alpha also attenuated relaxation responsiveness to isoproterenol but not to PGE2 or forskolin. TNF-alpha modulatory effects on GPCR-induced ASM responsiveness were completely abrogated by pertussis toxin, an inhibitor of Gialpha proteins. Taken together, these data suggest that TNF-alpha may participate in the development of airway hyperresponsiveness in asthma via the modulation of ASM responsiveness to both contractile and beta-adrenoceptor GPCR agonists. PMID- 12730148 TI - Validity of fluorescent microspheres method for bone blood flow measurement during intentional arterial hypotension. AB - In this study, we compared bone blood flow values obtained by simultaneously injected fluorescent (FM) and radiolabeled microspheres (RM) at stepwise reduced arterial blood pressure. Ten anesthetized female New Zealand White rabbits received simultaneous left ventricular injections of FM and RM at 90, 70, and 50 mmHg mean arterial blood pressure (MAP). After the experiments, both kidneys and long bones of all four limbs were removed and dissected in a standardized manner. Radioactivity (corrected for decay, background, and spillover) and fluorescence were determined, and blood flow values were calculated. Relative blood flow values estimated for each bone sample by RM and FM were significantly correlated (r = 0.98, slope = 0.99, and intercept = 0.04 for 90 mmHg; r = 0.98, slope = 0.94, and intercept = 0.09 for 70 mmHg; r = 0.98, slope = 0.96, and intercept = 0.07 for 50 mmHg). Blood flow values (ml x min-1 x 100 g-1) of right and left bone samples determined at the different arterial blood pressures were identical. During moderate hypotension (70 mmHg MAP), blood flow in all bone samples remained unchanged compared with 90 mmHg MAP, whereas a significant decrease of bone blood flow was observed at severe hypotension (50 mmHg MAP). Our results demonstrate that the FM technique is valid for measuring bone blood flow. Differences in bone blood flow during altered hemodynamic conditions can be detected reliably. In addition, changes in bone blood flow during hypotension indicate that vasomotor control mechanisms, as well as cardiac output, play a role in setting bone blood flow. PMID- 12730150 TI - nCPAP improves abnormal autonomic function in at-risk-for-SIDS infants with OSA. AB - We evaluated cardiovascular autonomic control and arousability during sleep in infants with obstructive sleep apnea (OSA) before and after 10 +/- 4 (mean +/- SD) days of treatment with nasal continuous positive airway pressure (nCPAP). Six OSA infants and 12 age-matched control infants were studied with polygraphic sleep studies at the age of 13 +/- 4 wk. During the study, 45 degrees head-up tilt tests were performed in slow-wave and rapid eye movement sleep. Blood pressure (BP) and heart rate (HR) were continuously monitored. All OSA infants had decreased initial BP and HR responses, followed by hypotension in two and hypertension in two. OSA infants displayed higher arousal thresholds in response to the tilt in rapid eye movement sleep (P < 0.005) and higher baseline HR (P < 0.05) than controls. nCPAP treatment normalized BP and HR responses as well as arousal thresholds to tilting and stabilized HR levels. OSA in infants may be linked with cardiovascular autonomic control disturbances and decreased arousability during sleep. These defects are improved by control of OSA with nCPAP. PMID- 12730149 TI - Effects of rapid saline infusion on lung mechanics and airway responsiveness in humans. AB - Lung mechanics and airway responsiveness to methacholine (MCh) were studied in seven volunteers before and after a 20-min intravenous infusion of saline. Data were compared with those of a time point-matched control study. The following parameters were measured: 1-s forced expiratory volume, forced vital capacity, flows at 40% of control forced vital capacity on maximal (Vm(40)) and partial (Vp(40)) forced expiratory maneuvers, lung volumes, lung elastic recoil, lung resistance (Rl), dynamic elastance (Edyn), and within-breath resistance of respiratory system (Rrs). Rl and Edyn were measured during tidal breathing before and for 2 min after a deep inhalation and also at different lung volumes above and below functional residual capacity. Rrs was measured at functional residual capacity and at total lung capacity. Before MCh, saline infusion caused significant decrements of forced expiratory volume in 1 s, Vm(40), and Vp(40), but insignificantly affected lung volumes, elastic recoil, Rl, Edyn, and Rrs at any lung volume. Furthermore, saline infusion was associated with an increased response to MCh, which was not associated with significant changes in the ratio of Vm(40) to Vp(40). In conclusion, mild airflow obstruction and enhanced airway responsiveness were observed after saline, but this was not apparently due to altered elastic properties of the lung or inability of the airways to dilate with deep inhalation. It is speculated that it was likely the result of airway wall edema encroaching on the bronchial lumen. PMID- 12730151 TI - Antifibrotic effects of suramin in injured skeletal muscle after laceration. AB - Muscle injuries are very common in traumatology and sports medicine. Although muscle tissue can regenerate postinjury, the healing process is slow and often incomplete; complete recovery after skeletal muscle injury is hindered by fibrosis. Our studies have shown that decreased fibrosis could improve muscle healing. Suramin has been found to inhibit transforming growth factor (TGF)-beta1 expression by competitively binding to the growth factor receptor. We conducted a series of tests to determine the antifibrotic effects of suramin on muscle laceration injuries. Our results demonstrate that suramin (50 microg/ml) can effectively decrease fibroblast proliferation and fibrotic-protein expression (alpha-smooth muscle actin) in vitro. In vivo, direct injection of suramin (2.5 mg) into injured murine muscle resulted in effective inhibition of muscle fibrosis and enhanced muscle regeneration, which led to efficient functional muscle recovery. These results support our hypothesis that prevention of fibrosis could enhance muscle regeneration, thereby facilitating more efficient muscle healing. This study could significantly contribute to the development of strategies to promote efficient muscle healing and functional recovery. PMID- 12730152 TI - Age-specific modification of local cutaneous vasodilation by capsaicin-sensitive primary afferents. AB - Age-related changes in neurogenic vasodilation mediated by sensory nerves may alter local skin blood flow (SkBF) responses in older individuals. The purpose of this study was to determine the age-specific modification of cutaneous vasodilation by capsaicin-sensitive primary afferent (CSPA) nerves during local heating. Nine young (18-30 yr), eight middle-aged (40-55 yr), and eight older (65 80 yr) healthy men participated in the experiments. Two local-heating protocols (rapid and slow) were performed before and after 1 wk of capsaicin pretreatment (CP), used to desensitize CSPAs. All temperatures were below those that elicit pain. SkBF was measured with a laser-Doppler imager and indexed to percentage of maximal cutaneous vascular conductance (%CVCmax). CP caused a significant reduction in %CVCmax in the middle-aged and older groups during slow heating (P < 0.05), without affecting %CVCmax in the young group. During rapid heating, CP significantly reduced (53.9 +/- 4.4 vs. 74.4 +/- 7.4% CVCmax, P < 0.05), but did not abolish, the initial sensory nerve-mediated rise in SkBF in the young group. No significant effects of CP on SkBF were observed during rapid heating in the middle-aged or older groups. These results indicate that, with advanced age, CSPA activity is more important to the maximal SkBF response during prolonged local heating, whereas it has a reduced role in the initial SkBF peak elicited by rapid local heating. In summary, CSPA activity contributes modestly to the overall SkBF response to local heating in an age-specific manner. PMID- 12730153 TI - Effects of prolonged reduction in blood flow on submandibular secretory function in anesthetized sheep. AB - Submandibular vascular and secretory responses to parasympathetic chorda-lingual (C-L) stimulation were investigated in anesthetized sheep before, during, and after an intracarotid (ic) infusion of endothelin-1 (ET-1). Stimulation of the peripheral end of the C-L nerve at 4 and 8 Hz produced a frequency-dependent reduction in submandibular vascular resistance (SVR) associated with a frequency dependent increase in submandibular blood flow, salivary flow, and Na+, K+, and protein output from the gland. During stimulation at 4 Hz, ic ET-1 significantly increased SVR (P < 0.01), without significantly affecting either the aortic blood pressure or heart rate. Submandibular blood flow (SBF) was reduced by 48 +/- 4% and the flow of saliva by 50 +/- 1%. The effect on blood and salivary flow persisted for at least 30 min after the infusion of ET-1. The reduction in SBF was associated with a diminution in the output of Na+,K+, and protein in the saliva (P < 0.01). These effects persisted for 30 min after the infusion of ET-1 had been discontinued and were linearly related to the flow of plasma throughout. PMID- 12730154 TI - Recombinant plasma gelsolin infusion attenuates burn-induced pulmonary microvascular dysfunction. AB - Reduced plasma concentrations of the extracellular actin-binding proteins gelsolin and Gc-globulin correlate with pulmonary failure and death in humans after injury. The purpose of this study was to investigate the role of plasma gelsolin in the pathophysiology of inflammation-induced lung injury. We postulated that plasma gelsolin levels decrease at an early time point after burn injury and that the intravenous infusion of gelsolin prevents burn-induced pulmonary microvascular dysfunction. Adult Sprague-Dawley rats were randomized to undergo a 40% body surface area thermal injury (Burn) or manipulation without burn (Sham). Plasma gelsolin and Gc-globulin concentrations were determined at various times during the first 6 days of injury by Western blotting. Other animals were randomized to receive either recombinant human gelsolin (0.078, 0.78, or 7.8 mg) or albumin (7.8 mg) before and 8 h after Burn or Sham. Twenty four hours later, pulmonary microvascular permeability was assessed by measuring the capillary filtration by use of an isolated, perfused lung model. We found that plasma gelsolin levels of burn-injured rats decreased to 10% of normal levels within 12 h and remained below normal levels for up to 6 days postinjury. Gc-globulin values also fall, but to a lesser extent and only transiently. Treatment of burned animals with intravenous infusions of recombinant human gelsolin prevented the increase in pulmonary microvascular permeability that accompanies this injury. Our findings are consistent with the hypothesis that plasma gelsolin depletion contributes to the pathophysiology of pulmonary microvascular dysfunction during inflammation. PMID- 12730155 TI - Roses by other names: taxonomy of the Rhizobiaceae. PMID- 12730156 TI - Methylotrophy in Methylobacterium extorquens AM1 from a genomic point of view. PMID- 12730158 TI - Attenuation of Mycobacterium tuberculosis by disruption of a mas-like gene or a chalcone synthase-like gene, which causes deficiency in dimycocerosyl phthiocerol synthesis. AB - Tuberculosis is one of the leading preventable causes of death. Emergence of drug resistant tuberculosis makes the discovery of new targets for antimycobacterial drugs critical. The unique mycobacterial cell wall lipids are known to play an important role in pathogenesis, and therefore the genes responsible for their biosynthesis offer potential new targets. To assess the possible role of some of the genes potentially involved in cell wall lipid synthesis, we disrupted a mas like gene, msl7, and a chalcone synthase-like gene, pks10, with phage-mediated delivery of the disruption construct, in which the target gene was disrupted by replacement of an internal segment with the hygromycin resistance gene (hyg). Gene disruption by allelic exchange in the case of each disruptant was confirmed by PCR and Southern blot analyses. Neither msl7 nor pks10 mutants could produce dimycocerosyl phthiocerol, although both could produce mycocerosic acids. Thus, it is concluded that these gene products are involved in the biosynthesis of phthiocerol. Both mutants were found to be attenuated in a murine model, supporting the hypothesis that dimycocerosyl phthiocerol is a virulence factor and thus the many steps involved in its biosynthesis offer potential novel targets for antimycobacterial therapy. PMID- 12730157 TI - Discordant phylogenies within the rrn loci of Rhizobia. AB - It is evident from complete genome sequencing results that lateral gene transfer and recombination are essential components in the evolutionary process of bacterial genomes. Since this has important implications for bacterial systematics, the primary objective of this study was to compare estimated evolutionary relationships among a representative set of alpha-Proteobacteria by sequencing analysis of three loci within their rrn operons. Tree topologies generated with 16S rRNA gene sequences were significantly different from corresponding trees assembled with 23S rRNA gene and internally transcribed space region sequences. Besides the incongruence in tree topologies, evidence that distinct segments along the 16S rRNA gene sequences of bacteria currently classified within the genera Bradyrhizobium, Mesorhizobium and Sinorhizobium have a reticulate evolutionary history was also obtained. Our data have important implications for bacterial taxonomy, because currently most taxonomic decisions are based on comparative 16S rRNA gene sequence analysis. Since phylogenetic placement based on 16S rRNA gene sequence divergence perhaps is questionable, we suggest that the proposals of bacterial nomenclature or changes in their taxonomy that have been made may not necessarily be warranted. Accordingly, a more conservative approach should be taken in the future, in which taxonomic decisions are based on the analysis of a wider variety of loci and comparative analytical methods are used to estimate phylogenetic relationships among the genomes under consideration. PMID- 12730159 TI - Mutational analysis of the carboxy-terminal (YGX)4 repeat domain of CpsD, an autophosphorylating tyrosine kinase required for capsule biosynthesis in Streptococcus pneumoniae. AB - In Streptococcus pneumoniae, CpsB, CpsC, and CpsD are essential for encapsulation, and mutants containing deletions of cpsB, cpsC, or cpsD exhibit rough colony morphologies. CpsD is an autophosphorylating protein-tyrosine kinase, CpsC is required for CpsD tyrosine phosphorylation, and CpsB is a phosphotyrosine-protein phosphatase. We have previously shown that autophosphorylation of CpsD at tyrosine attenuates its activity and consequently reduces the level of encapsulation and negatively regulates CPS production. In this study, we further investigated the role of the carboxy-terminal (YGX)(4) repeat domain of CpsD in encapsulation. A CpsD truncation mutant in which the entire (YGX)(4) repeat domain was removed was indistinguishable from a strain in which the entire cpsD gene had been deleted, indicating that the carboxy-terminal (YGX)(4) tail is required for CpsD activity in capsular polysaccharide production. Double mutants having a single tyrosine residue at position 2, 3, or 4 in the (YGX)(4) repeat domain and lacking CpsB exhibited a rough colony morphology, indicating that in the absence of an active protein-tyrosine phosphatase, phosphorylation of just one of the tyrosine residues in the (YGX)(4) repeat was sufficient to inactivate CpsD. When various mutants in which CpsD had either one or combinations of two or three tyrosine residues in the (YGX)(4) repeat domain were examined, only those with three tyrosine residues in the (YGX)(4) repeat domain were indistinguishable from the wild-type strain. The mutants with either one or two tyrosine residues exhibited mucoid colony morphologies. Further analysis of the mucoid strains indicated that the mucoid phenotype was not due to overproduction of capsular polysaccharide, as these strains actually produced less capsular polysaccharide than the wild-type strain. Thus, the tyrosine residues in the (YGX)(4) repeat domain are essential for normal functioning of CpsD. PMID- 12730160 TI - Roles of DegP in prevention of protein misfolding in the periplasm upon overexpression of penicillin acylase in Escherichia coli. AB - Enhancement of the production of soluble recombinant penicillin acylase in Escherichia coli via coexpression of a periplasmic protease/chaperone, DegP, was demonstrated. Coexpression of DegP resulted in a shift of in vivo penicillin acylase (PAC) synthesis flux from the nonproductive pathway to the productive one when pac was overexpressed. The number of inclusion bodies, which consist primarily of protein aggregates of PAC precursors in the periplasm, was highly reduced, and the specific PAC activity was highly increased. DegP was a heat shock protein induced in response to pac overexpression, suggesting that the protein could possibly suppress the physiological toxicity caused by pac overexpression. Coexpression of DegP(S210A), a DegP mutant without protease activity but retaining chaperone activity, could not suppress the physiological toxicity, suggesting that DegP protease activity was primarily responsible for the suppression, possibly by degradation of abnormal proteins when pac was overexpressed. However, a shortage of periplasmic protease activity was not the only reason for the deterioration in culture performance upon pac overexpression because coexpression of a DegP-homologous periplasmic protease, DegQ or DegS, could not suppress the physiological toxicity. The chaperone activity of DegP is proposed to be another possible factor contributing to the suppression. PMID- 12730161 TI - S-adenosylmethionine transport in Rickettsia prowazekii. AB - Rickettsia prowazekii, the causative agent of epidemic typhus, is an obligate, intracellular, parasitic bacterium that grows within the cytoplasm of eucaryotic host cells. Rickettsiae exploit this intracellular environment by using transport systems for the compounds available in the host cell's cytoplasm. Analysis of the R. prowazekii Madrid E genome sequence revealed the presence of a mutation in the rickettsial metK gene, the gene encoding the enzyme responsible for the synthesis of S-adenosylmethionine (AdoMet). Since AdoMet is required for rickettsial processes, the apparent inability of this strain to synthesize AdoMet suggested the presence of a rickettsial AdoMet transporter. We have confirmed the presence of an AdoMet transporter in the rickettsiae which, to our knowledge, is the first bacterial AdoMet transporter identified. The influx of AdoMet into rickettsiae was a saturable process with a K(T) of 2.3 micro M. Transport was inhibited by S adenosylethionine and S-adenosylhomocysteine but not by sinfungin or methionine. Transport was also inhibited by 2,4-dinitrophenol, suggesting an energy-linked transport mechanism, and by N-ethylmaleimide. AdoMet transporters with similar properties were also identified in the Breinl strain of R. prowazekii and in Rickettsia typhi. By screening Escherichia coli clone banks for AdoMet transport, the R. prowazekii gene coding for a transporter, RP076 (sam), was identified. AdoMet transport in E. coli containing the R. prowazekii sam gene exhibited kinetics similar to that seen in rickettsiae. The existence of a rickettsial transporter for AdoMet raises intriguing questions concerning the evolutionary relationship between the synthesis and transport of this essential metabolite. PMID- 12730162 TI - Leucines 193 and 194 at the N-terminal domain of the XylS protein, the positive transcriptional regulator of the TOL meta-cleavage pathway, are involved in dimerization. AB - Members of the AraC/XylS family of transcriptional regulators are usually organized in two domains: a conserved domain made up of 100 amino acids and frequently located at the C-terminal end, involved in DNA binding; and an N terminal nonconserved domain involved in signal recognition, as is the case for regulators involved in the control of carbon metabolism (R. Tobes and J. L. Ramos, Nucleic Acids Res. 30:318-321, 2002). The XylS protein, which is extremely insoluble, controls expression of the meta-cleavage pathway for alkylbenzoate metabolism. We fused the N-terminal end of XylS to the maltose-binding protein (MBP) in vitro and found in glutaraldehyde cross-linking assays that the protein dimerized. Experiments with a chimeric N-terminal XylS linked to a 'LexA protein showed that the dimer was stabilized in the presence of alkylbenzoates. Sequence alignments with AraC and UreR allowed us to identify three residues, Leu193, Leu194, and Ile205, as potentially being involved in dimerization. Site-directed mutagenesis of XylS in which each of the above residues was replaced with Ala revealed that Leu193 and Leu194 were critical for activity and that a chimera in which LexA was linked to the N terminus of XylSLeu193Ala or XylSLeu194Ala was not functional. Dimerization of the chimeras MBP-N-XylSLeu193Ala and MBP-N XylSLeu194Ala was not observed in cross-linking assays with glutaraldehyde. PMID- 12730163 TI - Regulation of the cellulosomal CelS (cel48A) gene of Clostridium thermocellum is growth rate dependent. AB - Clostridium thermocellum produces an extracellular multienzyme complex, termed cellulosome, that allows efficient solubilization of crystalline cellulose. One of the major enzymes in this complex is the CelS (Cel48A) exoglucanase. The regulation of CelS at the protein and transcriptional levels was studied using batch and continuous cultures. The results of sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot analyses indicated that the amount of CelS in the supernatant fluids of cellobiose-grown cultures is lower than that of cellulose-grown cultures. The transcriptional level of celS mRNA was determined quantitatively by RNase protection assays with batch and continuous cultures under carbon and nitrogen limitation. The amount of celS mRNA transcripts per cell was about 180 for cells grown under carbon limitation at growth rates of 0.04 to 0.21 h(-1) and 80 and 30 transcripts per cell for batch cultures at growth rates of 0.23 and 0.35 h(-1), respectively. Under nitrogen limitation, the corresponding levels were 110, 40, and 30 transcripts/cell for growth rates of 0.07, 0.11, and 0.14 h(-1), respectively. Two major transcriptional start sites were detected at positions -140 and -145 bp, upstream of the translational start site of the celS gene. The potential promoters exhibited homology to known sigma factors (i.e., sigma(A) and sigma(B)) of Bacillus subtilis. The relative activity of the two promoters remained constant under the conditions studied and was in agreement with the results of the RNase protection assay, in which the observed transcriptional activity was inversely proportional to the growth rate. PMID- 12730164 TI - Synthesis of catalytically active form III ribulose 1,5-bisphosphate carboxylase/oxygenase in archaea. AB - Ribulose 1,5 bisphosphate carboxylase/oxygenase (RubisCO) catalyzes the biological reduction and assimilation of carbon dioxide gas to organic carbon; it is the key enzyme responsible for the bulk of organic matter found on earth. Until recently it was believed that there are only two forms of RubisCO, form I and form II. However, the recent completion of several genome-sequencing projects uncovered open reading frames resembling RubisCO in the third domain of life, the archaea. Previous work and homology comparisons suggest that these enzymes represent a third form of RubisCO, form III. While earlier work indicated that two structurally distinct recombinant archaeal RubisCO proteins catalyzed bona fide RubisCO reactions, it was not established that the rbcL genes of anaerobic archaea can be transcribed and translated to an active enzyme in the native organisms. In this report, it is shown not only that Methanococcus jannaschii, Archaeoglobus fulgidus, Methanosarcina acetivorans, and Methanosarcina barkeri possess open reading frames with the residues required for catalysis but also that the RubisCO protein from these archaea accumulates in an active form under normal growth conditions. In addition, the form III RubisCO gene (rbcL) from M. acetivorans was shown to complement RubisCO deletion strains of Rhodobacter capsulatus and Rhodobacter sphaeroides under both photoheterotrophic and photoautotrophic growth conditions. These studies thus indicate for the first time that archaeal form III RubisCO functions in a physiologically significant fashion to fix CO(2). Furthermore, recombinant M. jannaschii, M. acetivorans, and A. fulgidus RubisCO possess unique properties with respect to quaternary structure, temperature optima, and activity in the presence of molecular oxygen compared to the previously described Thermococcus kodakaraensis and halophile proteins. PMID- 12730165 TI - Characterization of the double-partitioning modules of R27: correlating plasmid stability with plasmid localization. AB - Plasmid R27 contains two independent partitioning modules, designated Par1 and Par2, within transfer region 2. Par1 is member of the type I partitioning family (Walker-type ATPase), and Par2 is a member of the type II partitioning family (actin-type ATPase). Stability tests of cloned Par1 and Par2 and insertional disruptions of Par1 and Par2 within R27 demonstrated that Par1 is the major stability determinant whereas Par2 is the minor stability determinant. Creation of double-partitioning mutants resulted in R27 integrating into the chromosome, suggesting that at least one partitioning module is required for R27 to exist in the extrachromosomal form. Using the lacO/LacI-green fluorescent protein (GFP) system, we labeled and visualized R27 and R27 partitioning mutants (Par1(-) and Par2(-)) under different growth conditions in live Escherichia coli cells. Plasmid R27 was visualized as the discrete GFP foci present at the mid- and quarter-cell regions in >99% of the cells. Time lapse experiments demonstrated that an increase in R27 plasmid foci resulted from focus duplication in either the mid- or quarter-cell regions of E. coli. Both R27 Par(-) variants gave a high percentage of plasmidless cells, as suggested by a uniform GFP signal, and cells with GFP patterns scattered throughout the entire cell, suggesting that plasmid molecules are randomly distributed throughout the cytoplasm. Those cells that did contain R27 Par(-) with one or two discrete foci had localization patterns that were statistically different from those formed with wild-type R27. Therefore, these results suggest that partitioning-impaired plasmids are characterized by individual and clustered plasmids that are randomly located within the host cytoplasm. PMID- 12730167 TI - The Escherichia coli Fis protein stimulates bacteriophage lambda integrative recombination in vitro. AB - The Escherichia coli nucleoid-associated protein Fis was previously shown to be involved in bacteriophage lambda site-specific recombination in vivo, enhancing the levels of both integrative recombination and excisive recombination. While purified Fis protein was shown to stimulate in vitro excision, Fis appeared to have no effect on in vitro integration reactions even though a 15-fold drop in lysogenization frequency had previously been observed in fis mutants. We demonstrate here that E. coli Fis protein does stimulate integrative lambda recombination in vitro but only under specific conditions which likely mimic natural in vivo recombination more closely than the standard conditions used in vitro. In the presence of suboptimal concentrations of Int protein, Fis stimulates the rate of integrative recombination significantly. In addition, Fis enhances the recombination of substrates with nonstandard topologies which may be more relevant to the process of in vivo phage lambda recombination. These data support the hypothesis that Fis may play an essential role in lambda recombination in the host cell. PMID- 12730166 TI - Cell-cycle-regulated expression and subcellular localization of the Caulobacter crescentus SMC chromosome structural protein. AB - Structural maintenance of chromosomes proteins (SMCs) bind to DNA and function to ensure proper chromosome organization in both eukaryotes and bacteria. Caulobacter crescentus possesses a single SMC homolog that plays a role in organizing and segregating daughter chromosomes. Approximately 1,500 to 2,000 SMC molecules are present per cell during active growth, corresponding to one SMC complex per 6,000 to 8,000 bp of chromosomal DNA. Although transcription from the smc promoter is induced during early S phase, a cell cycle transcription pattern previously observed with multiple DNA replication and repair genes, the SMC protein is present throughout the entire cell cycle. Examination of the intracellular location of SMC showed that in swarmer cells, which do not replicate DNA, the protein forms two or three foci. Stalked cells, which are actively engaged in DNA replication, have three or four SMC foci per cell. The SMC foci appear randomly distributed in the cell. Many predivisional cells have bright polar SMC foci, which are lost upon cell division. Thus, chromosome compaction likely involves dynamic aggregates of SMC bound to DNA. The aggregation pattern changes as a function of the cell cycle both during and upon completion of chromosome replication. PMID- 12730168 TI - Identification of rocA, a positive regulator of covR expression in the group A streptococcus. AB - In the group A streptococcus (GAS; Streptococcus pyogenes), a two-component system known as CovRS (or CsrRS) regulates about 15% of the genes, including several important virulence factors like the hyaluronic acid capsule. Most of these genes, including covR itself, are negatively regulated by CovR. We have isolated two independent ISS1 insertions in an open reading frame (ORF) that increases CovR expression as measured by a Pcov-gusA reporter fusion in single copy in the GAS chromosome. This ORF, named rocA for "regulator of Cov," activates covR transcription about threefold. As expected, a rocA mutant is mucoid and produces more transcript from the has promoter since this promoter is repressed by CovR. This effect is dependent on the presence of a wild-type covR gene. In contrast to its activation of Pcov, RocA negatively regulates its own expression. This autoregulation is not dependent on the presence of the covR gene. All the phenotypes of the rocA mutant were complemented by the presence of the rocA gene on a plasmid. The rocA gene is present in strains of all nine M serotypes of GAS tested and is absent from strains representing 11 other groups of streptococci and related bacteria, including strains of the closely related group C and G streptococci. It seems likely that rocA plays an important role in the pathogenesis of GAS since it affects expression of the global regulator CovR. PMID- 12730169 TI - PaeX, a second pectin acetylesterase of Erwinia chrysanthemi 3937. AB - Erwinia chrysanthemi causes soft-rot diseases of various plants by enzymatic degradation of the pectin in plant cell walls. Pectin is a complex polysaccharide. The main chain is constituted of galacturonate residues, and some of them are modified by methyl and/or acetyl esterification. Esterases are necessary to remove these modifications and, thus, to facilitate the further degradation of the polysaccharidic chain. In addition to PaeY, the first pectin acetylesterase identified in the E. chrysanthemi strain 3937, we showed that this bacterium produces a second pectin acetylesterase encoded by the gene paeX. The paeX open reading frame encodes a 322-residue precursor protein of 34,940 Da, including a 21-amino-acid signal peptide. Analysis of paeX transcription, by using gene fusions, revealed that it is induced by pectic catabolic products and affected by catabolite repression. The expression of paeX is regulated by the repressor KdgR, which controls all the steps of pectin catabolism; by the repressor PecS, which controls most of the pectinase genes; and by catabolite regulatory protein, the global activator of sugar catabolism. The paeX gene is situated in a cluster of genes involved in the catabolism and transport of pectic oligomers. In induced conditions, the two contiguous genes kdgM, encoding an oligogalacturonate-specific porin, and paeX are both transcribed as an operon from a promoter proximal to kdgM, but transcription of paeX can also be uncoupled from that of kdgM in noninduced conditions. PaeX is homologous to the C-terminal domain of the Butyrivibrio fibriosolvens xylanase XynB and to a few bacterial esterases. PaeX contains the typical box (GxSxG) corresponding to the active site of the large family of serine hydrolases. Purified PaeX releases acetate from various synthetic substrates and from sugar beet pectin. The PaeX activity increased after previous depolymerization and demethylation of pectin, indicating that its preferred substrates are nonmethylated oligogalacturonides. PaeX is mostly found in the periplasmic space of E. chrysanthemi. These data suggest that PaeX is mainly involved in the deacetylation of esterified oligogalacturonides that enter the periplasm by the KdgM porin. PMID- 12730170 TI - Repair system for noncanonical purines in Escherichia coli. AB - Exposure of Escherichia coli strains deficient in molybdopterin biosynthesis (moa) to the purine base N-6-hydroxylaminopurine (HAP) is mutagenic and toxic. We show that moa mutants exposed to HAP also exhibit elevated mutagenesis, a hyperrecombination phenotype, and increased SOS induction. The E. coli rdgB gene encodes a protein homologous to a deoxyribonucleotide triphosphate pyrophosphatase from Methanococcus jannaschii that shows a preference for purine base analogs. moa rdgB mutants are extremely sensitive to killing by HAP and exhibit increased mutagenesis, recombination, and SOS induction upon HAP exposure. Disruption of the endonuclease V gene, nfi, rescues the HAP sensitivity displayed by moa and moa rdgB mutants and reduces the level of recombination and SOS induction, but it increases the level of mutagenesis. Our results suggest that endonuclease V incision of DNA containing HAP leads to increased recombination and SOS induction and even cell death. Double-strand break repair mutants display an increase in HAP sensitivity, which can be reversed by an nfi mutation. This suggests that cell killing may result from an increase in double strand breaks generated when replication forks encounter endonuclease V-nicked DNA. We propose a pathway for the removal of HAP from purine pools, from deoxynucleotide triphosphate pools, and from DNA, and we suggest a general model for excluding purine base analogs from DNA. The system for HAP removal consists of a molybdoenzyme, thought to detoxify HAP, a deoxyribonucleotide triphosphate pyrophosphatase that removes noncanonical deoxyribonucleotide triphosphates from replication precursor pools, and an endonuclease that initiates the removal of HAP from DNA. PMID- 12730171 TI - Swarm-cell differentiation in Salmonella enterica serovar typhimurium results in elevated resistance to multiple antibiotics. AB - Although a wealth of knowledge exists about the molecular and biochemical mechanisms governing the swimming motility of Salmonella enterica serovar Typhimurium, its surface swarming behavior has not been extensively characterized. When inoculated onto a semisolid agar medium supplemented with appropriate nutrients, serovar Typhimurium undergoes a morphological differentiation whereby single cells hyperflagellate and elongate into nonseptate, multinucleate swarm cells. Swarm migration is a collective behavior of groups of cells. We have isolated a MudJ insertion mutant of serovar Typhimurium 14028 that failed to swarm under any conditions. The site of the MudJ insertion was determined to be in the pmrK locus within the pmrHFIJKLM operon, which was previously demonstrated to confer resistance to cationic antimicrobial peptides. beta-Galactosidase assays, using the pmrK::lacZ transcriptional fusion, showed increased expression of the pmr operon in swarm cells compared to that in vegetative cells. In concurrence with the expression data, swarm cells exhibited greater tolerance to polymyxin. To compare the profiles of vegetative and swarm cell resistance to other antibiotics, E-test strips representing a wide range of antibiotic classes were used. Swarm cells exhibited elevated resistance to a variety of antibiotics, including those that target the cell envelope, protein translation, DNA replication, and transcription. These observations, in addition to the dramatic morphological changes associated with the swarming phenotype, provide an intriguing model for examining global differences between the physiological states of vegetative and swarm cells of serovar Typhimurium. PMID- 12730172 TI - CodY is a nutritional repressor of flagellar gene expression in Bacillus subtilis. AB - Expression of the sigma(D)-dependent flagellin gene, hag, is repressed by the CodY protein in nutrient-rich environments. Analysis of a codY mutant bearing a hag-lacZ reporter suggests that the availability of amino acids in the environment is the specific signal that triggers this repression. Further, hag lacZ expression appears to be sensitive to intracellular GTP levels, as demonstrated by increased expression upon addition of decoyinine. This result is consistent with the postulate that the availability of amino acids in the environment effects intracellular GTP levels through the stringent response. However, the levels of hag-lacZ measured upon the addition of subsets of amino acids suggest an additional mechanism(s). CodY is a DNA binding protein that could repress flagellin expression directly by binding to the hag promoter region, or indirectly by binding to the fla/che promoter region that governs expression of the sigma(D) transcriptional activator required for hag gene expression. Using an electrophoretic mobility shift assay, we have demonstrated that purified CodY protein binds specifically to both the hag and fla/che promoter fragments. Additionally, CodY acts as a nutritional repressor of transcription from the fla/che promoter region that contains two functional promoters. CodY binds to both the sigma(D)- and sigma(A)-dependent promoters in this region, as demonstrated by DNase I footprint analyses. Footprint analyses of the hag gene demonstrated that CodY binds downstream of its sigma(D)-dependent promoter. Taken together, these results identify new members of the CodY regulon that encode motility functions in Bacillus subtilis and are controlled by the sigma(D) alternate sigma factor. PMID- 12730174 TI - Transcription regulation coupling of the divergent argG and metY promoters in Escherichia coli K-12. AB - The cAMP-catabolite activator protein (CAP) complex is a pleiotropic regulator that regulates a vast number of Escherichia coli genes, including those involved in carbon metabolism. We identified two new targets of this complex: argG, which encodes the arginosuccinate synthase involved in the arginine biosynthetic pathway, and metY, which encodes one of the two methionine tRNA initiators, tRNAf2Met. The cAMP-CAP complex activates argG transcription and inhibits metY transcription from the same DNA position. We also show that ArgR, the specific repressor of the arginine biosynthetic pathway, together with its arginine cofactor, acts on the regulation of metY mediated by CAP. The regulation of the two divergent promoters is thus simultaneously controlled not only by the cAMP CAP complex, a global regulator, but also by a specific regulator of arginine metabolism, suggesting a previously unsuspected link between carbon metabolism and translation initiation. PMID- 12730173 TI - Characterization of NorR protein, a multifunctional regulator of norA expression in Staphylococcus aureus. AB - We characterized a Staphylococcus aureus norA gene expression regulator, NorR, initially identified from its binding to the norA promoter. The norR gene was 444 bp in length, located approximately 7 kb upstream from the norA gene, and encoded a predicted 17.6-kDa protein. Overexpression of norR in wild-type S. aureus strain ISP794 led to a fourfold decrease in sensitivity to quinolones and ethidium bromide and an increase in the level of norA transcripts, suggesting that NorR acts as a positive regulator of norA expression. Overexpression of norR in sarA and agr mutants did not alter quinolone sensitivity or levels of norA transcription, indicating that the presence of these two global regulatory systems is necessary for NorR to affect the expression of norA. Insertion and disruption of norR in ISP794 increased resistance to quinolones by 4- to 16-fold but had no effect on norA transcription, suggesting that NorR acts as a repressor for another unidentified efflux pump or pumps. These mutants also exhibited an exaggerated clumping phenotype in liquid media, which was complemented fully by a plasmid-encoded norR gene. Collectively, these results indicate that NorR is a multifunctional regulator, affecting cell surface properties as well as the expression of NorA and likely other multidrug resistance efflux pumps. PMID- 12730175 TI - Early colonization events in the mutualistic association between Steinernema carpocapsae nematodes and Xenorhabdus nematophila bacteria. AB - The bacterium Xenorhabdus nematophila is a mutualist of the entomopathogenic nematode Steinernema carpocapsae. During its life cycle, the bacterium exists both separately from the nematode and as an intestinal resident of a nonfeeding nematode form, the infective juvenile (IJ). The progression of X. nematophila from an ex vivo existence to a specific and persistent colonization of IJs is a model to understand the mechanisms mediating the initiation and maintenance of benign host-microbe interactions. To help characterize this process, we constructed an X. nematophila strain that constitutively expresses green fluorescent protein, which allowed its presence to be monitored within IJs. Using this strain, we showed that few bacterial cells initiate colonization of an individual IJ and that these grow inside the lumen of the IJ intestine in a reproducible polyphasic pattern during colonization. In accordance with these two observations, we demonstrated that the final population of bacteria in a nematode is of predominantly monoclonal origin, suggesting that only one or two bacterial clones initiate or persist during colonization of an individual nematode. These data suggest that X. nematophila initiates IJ colonization by competing for limited colonization sites or resources within the nematode intestine. This report represents the first description of the biological interactions occurring between X. nematophila and S. carpocapsae during the early stages of the colonization process, provides insights into the physiology of X. nematophila in its host niche, and will facilitate interpretation of future data regarding the molecular events mediating this process. PMID- 12730176 TI - Characterization of the Xanthomonas axonopodis pv. glycines Hrp pathogenicity island. AB - We sequenced an approximately 29-kb region from Xanthomonas axonopodis pv. glycines that contained the Hrp type III secretion system, and we characterized the genes in this region by Tn3-gus mutagenesis and gene expression analyses. From the region, hrp (hypersensitive response and pathogenicity) and hrc (hrp and conserved) genes, which encode type III secretion systems, and hpa (hrp associated) genes were identified. The characteristics of the region, such as the presence of many virulence genes, low G+C content, and bordering tRNA genes, satisfied the criteria for a pathogenicity island (PAI) in a bacterium. The PAI was composed of nine hrp, nine hrc, and eight hpa genes with seven plant inducible promoter boxes. The hrp and hrc mutants failed to elicit hypersensitive responses in pepper plants but induced hypersensitive responses in all tomato plants tested. The Hrp PAI of X. axonopodis pv. glycines resembled the Hrp PAIs of other Xanthomonas species, and the Hrp PAI core region was highly conserved. However, in contrast to the PAI of Pseudomonas syringae, the regions upstream and downstream from the Hrp PAI core region showed variability in the xanthomonads. In addition, we demonstrate that HpaG, which is located in the Hrp PAI region of X. axonopodis pv. glycines, is a response elicitor. Purified HpaG elicited hypersensitive responses at a concentration of 1.0 micro M in pepper, tobacco, and Arabidopsis thaliana ecotype Cvi-0 by acting as a type III secreted effector protein. However, HpaG failed to elicit hypersensitive responses in tomato, Chinese cabbage, and A. thaliana ecotypes Col-0 and Ler. This is the first report to show that the harpin-like effector protein of Xanthomonas species exhibits elicitor activity. PMID- 12730177 TI - Energy-generating enzymes of Burkholderia cepacia and their interactions with macrophages. AB - We previously demonstrated that several clinical and environmental isolates of Burkholderia cepacia secreted ATP-utilizing enzymes to the medium; the secretion of these enzymes by cystic fibrosis lung isolate strain 38 was shown to be greatly enhanced in the presence of alpha(2)-macroglobulin. Fractionation of the growth medium of cystic fibrosis isolate strain 71 belonging to genomovar I demonstrated the presence of two additional proteins, homologues of Pseudomonas aeruginosa azurin and cytochrome c(551), which are normally involved in electron transfer during denitrification. A Q-Sepharose column flowthrough fraction of the growth medium of B. cepacia strain 71 enriched with the azurin and cytochrome c(551) homologues triggered apoptosis in macrophages and mast cells, leading to their death. Incubation of the Q-Sepharose column flowthrough fraction with antiazurin and anti-cytochrome c(551) antibodies greatly reduced cell death. We cloned and hyperexpressed a gene from B. cepacia strain 71 that encodes the homologue of P. aeruginosa azurin. Such azurin homologues were detected in the growth medium of several strains belonging to genomovars I, III, and VI but not in the growth medium of strains belonging to other genomovars. The growth medium of the strains that elaborated the azurin homologue had high cytotoxicity towards macrophages. Purified azurin homologue was shown to induce apoptosis in macrophages in a caspase-dependent manner and was localized in both the cytosol and nucleus when incubated with or microinjected into macrophages. This is an interesting example of the interaction of a bacterial protein normally involved in cellular energetics with macrophages to effect their cell death. PMID- 12730178 TI - Global stage-specific gene regulation during the developmental cycle of Chlamydia trachomatis. AB - Distinct morphological changes associated with the complex development cycle of the obligate intracellular bacterial pathogen Chlamydia trachomatis have been historically well characterized by microscopy. A number of temporally regulated genes have been characterized previously, suggesting that the chlamydial developmental cycle is regulated at the transcriptional level. This hypothesis was tested by microarray analysis in which the entire C. trachomatis genome was analyzed, providing a comprehensive assessment of global gene regulation throughout the chlamydial developmental cycle. Seven temporally cohesive gene clusters were identified, with 22% (189 genes) of the genome differentially expressed during the developmental cycle. The correlation of these gene clusters with hallmark morphological events of the chlamydial developmental cycle suggests three global stage-specific networks of gene regulation. PMID- 12730179 TI - Genes of the GadX-GadW regulon in Escherichia coli. AB - Acid in the stomach is thought to be a barrier to bacterial colonization of the intestine. Escherichia coli, however, has three systems for acid resistance, which overcome this barrier. The most effective of these systems is dependent on transport and decarboxylation of glutamate. GadX regulates two genes that encode isoforms of glutamate decarboxylase critical to this system, but additional genes associated with the glutamate-dependent acid resistance system remained to be identified. The gadX gene and a second downstream araC-like transcription factor gene, gadW, were mutated separately and in combination, and the gene expression profiles of the mutants were compared to those of the wild-type strain grown in neutral and acidified media under conditions favoring induction of glutamate dependent acid resistance. Cluster and principal-component analyses identified 15 GadX-regulated, acid-inducible genes. Reverse transcriptase mapping demonstrated that these genes are organized in 10 operons. Analysis of the strain lacking GadX but possessing GadW confirmed that GadX is a transcriptional activator under acidic growth conditions. Analysis of the strain lacking GadW but possessing GadX indicated that GadW exerts negative control over three GadX target genes. The strain lacking both GadX and GadW was defective in acid induction of most but not all GadX target genes, consistent with the roles of GadW as an inhibitor of GadX dependent activation of some genes and an activator of other genes. Resistance to acid was decreased under certain conditions in a gadX mutant and even more so by combined mutation of gadX and gadW. However, there was no defect in colonization of the streptomycin-treated mouse model by the gadX mutant in competition with the wild type, and the gadX gadW mutant was a better colonizer than the wild type. Thus, E. coli colonization of the mouse does not appear to require glutamate-dependent acid resistance. PMID- 12730181 TI - Characterization of the chlorate reductase from Pseudomonas chloritidismutans. AB - A chlorate reductase has been purified from the chlorate-reducing strain Pseudomonas chloritidismutans. Comparison with the periplasmic (per)chlorate reductase of strain GR-1 showed that the cytoplasmic chlorate reductase of P. chloritidismutans reduced only chlorate and bromate. Differences were also found in N-terminal sequences, molecular weight, and subunit composition. Metal analysis and electron paramagnetic resonance measurements showed the presence of iron and molybdenum, which are also found in other dissimilatory oxyanion reductases. PMID- 12730180 TI - Conservation of plasmid maintenance functions between linear and circular plasmids in Borrelia burgdorferi. AB - The Lyme disease agent Borrelia burgdorferi maintains both linear and circular plasmids that appear to be essential for mammalian infection. Recent studies have characterized the circular plasmid regions that confer autonomous replication, but the genetic elements necessary for linear plasmid maintenance have not been experimentally identified. Two vectors derived from linear plasmids lp25 and lp28 1 were constructed and shown to replicate autonomously in B. burgdorferi. These vectors identify internal regions of linear plasmids necessary for autonomous replication in B. burgdorferi. Although derived from linear plasmids, the vectors are maintained in circular form in B. burgdorferi, indicating that plasmid maintenance functions are conserved, regardless of DNA form. Finally, derivatives of these vectors indicate that paralogous gene family 49 is apparently not required for either circular or linear plasmid replication. PMID- 12730182 TI - Alpha-toxin is required for biofilm formation by Staphylococcus aureus. AB - Staphylococcus aureus is a common pathogen associated with nosocomial infections. It can persist in clinical settings and gain increased resistance to antimicrobial agents through biofilm formation. We have found that alpha-toxin, a secreted, multimeric, hemolytic toxin encoded by the hla gene, plays an integral role in biofilm formation. The hla mutant was unable to fully colonize plastic surfaces under both static and flow conditions. Based on microscopy studies, we propose that alpha-hemolysin is required for cell-to-cell interactions during biofilm formation. PMID- 12730183 TI - Identification of the L-aspartate transporter in Bacillus subtilis. AB - YveA of Bacillus subtilis, a putative transporter of the amino acid/polyamine/organocation (APC) superfamily, is shown to mediate uptake of both L-aspartate and L-glutamate as well as having sensitivity to L-aspartate hydroxamate. This 14 TMS protein is the primary aspartate uptake system in B. subtilis and serves as the prototype for a new family within the APC superfamily. PMID- 12730184 TI - The single superoxide dismutase of Rhodobacter capsulatus is a cambialistic, manganese-containing enzyme. AB - The phototrophic bacterium Rhodobacter capsulatus contains a single, oxygen responsive superoxide dismutase (SOD(Rc)) homologous to iron-containing superoxide dismutase enzymes. Recombinant SOD(Rc), however, displayed higher activity after refolding with Mn(2+), especially when the pH of the assay mixture was raised. SOD(Rc) isolated from Rhodobacter cells also preferentially contains manganese, but metal discrimination depends on the culture conditions, with iron fractions increasing from 7% in aerobic cultures up to 40% in photosynthetic cultures. Therefore, SOD(Rc) behaves as a Mn-containing dismutase with cambialistic properties. PMID- 12730185 TI - The Bacillus subtilis acyl lipid desaturase is a delta5 desaturase. AB - Bacillus subtilis was recently reported to synthesize unsaturated fatty acids (UFAs) with a double bond at positions delta5, delta7, and delta9 (M. H. Weber, W. Klein, L. Muller, U. M. Niess, and M. A. Marahiel, Mol. Microbiol. 39:1321 1329, 2001). Since this finding would have considerable importance in the double bond positional specificity displayed by the B. subtilis acyl lipid desaturase, we have attempted to confirm this observation. We report that the double bond of UFAs synthesized by B. subtilis is located exclusively at the delta5 position, regardless of the growth temperature and the length chain of the fatty acids. PMID- 12730186 TI - Differential expression of the components of the two alkane hydroxylases from Pseudomonas aeruginosa. AB - Oxidation of n-alkanes in bacteria is normally initiated by an enzyme system formed by a membrane-bound alkane hydroxylase and two soluble proteins, rubredoxin and rubredoxin reductase. Pseudomonas aeruginosa strains PAO1 and RR1 contain genes encoding two alkane hydroxylases (alkB1 and alkB2), two rubredoxins (alkG1 and alkG2), and a rubredoxin reductase (alkT). We have localized the promoters for these genes and analyzed their expression under different conditions. The alkB1 and alkB2 genes were preferentially expressed at different moments of the growth phase; expression of alkB2 was highest during the early exponential phase, while alkB1 was induced at the late exponential phase, when the growth rate decreased. Both genes were induced by C(10) to C(22)/C(24) alkanes but not by their oxidation derivatives. However, the alkG1, alkG2, and alkT genes were expressed at constant levels in both the absence and presence of alkanes. PMID- 12730187 TI - The yggH gene of Escherichia coli encodes a tRNA (m7G46) methyltransferase. AB - We cloned, expressed, and purified the Escherichia coli YggH protein and show that it catalyzes the S-adenosyl-L-methionine-dependent formation of N(7) methylguanosine at position 46 (m(7)G46) in tRNA. Additionally, we generated an E. coli strain with a disrupted yggH gene and show that the mutant strain lacks tRNA (m(7)G46) methyltransferase activity. PMID- 12730188 TI - Controlled initiation of chromosomal replication in Escherichia coli requires functional Hda protein. AB - Regulatory inactivation of DnaA helps ensure that the Escherichia coli chromosome is replicated only once per cell cycle, through accelerated hydrolysis of active replication initiator ATP-DnaA to inactive ADP-DnaA. Analysis of deltahda strains revealed that the regulatory inactivation of DnaA component Hda is necessary for maintaining controlled initiation but not for cell growth or viability. PMID- 12730189 TI - Global conformations, hydrodynamics, and X-ray scattering properties of Taq and Escherichia coli DNA polymerases in solution. AB - Escherichia coli polymerase 1 (Pol 1) and Thermus aquaticus Taq polymerase are homologous Type I DNA polymerases, each comprised of a polymerase domain, a proofreading domain (inactive in Taq), and a 5' nuclease domain. "Klenow" and "Klentaq" are the large fragments of Pol 1 and Taq and are functional polymerases lacking the 5' nuclease domain. In the available crystal structures of full length Taq, the 5' nuclease domain is positioned in two different orientations: in one structure, it is extended out into solution, whereas in the other, it is folded up against the polymerase domain in a more compact structure. Analytical ultracentrifugation experiments report s20,w values of 5.05 for Taq, 4.1 for Klentaq, 5.3 for E. coli Pol 1, and 4.6 for Klenow. Measured partial specific volumes are all quite similar, indicating no significant differences in packing density between the mesophilic and thermophilic proteins. Small angle x-ray scattering studies report radii of gyration of 38.3 A for Taq, 30.7 A for Klentaq, and 30.5 A for Klenow. The hydrodynamic and x-ray scattering properties of the polymerases were also calculated directly from the different crystal structures using the programs HYDROPRO (Garcia De La Torre, J., Huertas, M. L., and Carrasco, B. (2000) Biophys J. 78, 719-730) and CRYSOL (Svergun, D. I., Barberato, C., and Koch, M. H. J. (1995) J. Appl. Crystalogr. 28, 768-773), respectively. The combined experimental and computational characterizations indicate that the full-length polymerases in solution are in a conformation where the 5' nuclease domain is extended into solution. Further, the radius of gyration, and hence the global conformation of Taq polymerase, is not altered by the binding of either matched primer template DNA or ddATP. PMID- 12730190 TI - Insights into the effects on metal binding of the systematic substitution of five key glutamate ligands in the ferritin of Escherichia coli. AB - Ferritins are nearly ubiquitous iron storage proteins playing a fundamental role in iron metabolism. They are composed of 24 subunits forming a spherical protein shell encompassing a central iron storage cavity. The iron storage mechanism involves the initial binding and subsequent O2-dependent oxidation of two Fe2+ ions located at sites A and B within the highly conserved dinuclear "ferroxidase center" in individual subunits. Unlike animal ferritins and the heme-containing bacterioferritins, the Escherichia coli ferritin possesses an additional iron binding site (site C) located on the inner surface of the protein shell close to the ferroxidase center. We report the structures of five E. coli ferritin variants and their Fe3+ and Zn2+ (a redox-stable alternative for Fe2+) derivatives. Single carboxyl ligand replacements in sites A, B, and C gave unique effects on metal binding, which explain the observed changes in Fe2+ oxidation rates. Binding of Fe2+ at both A and B sites is clearly essential for rapid Fe2+ oxidation, and the linking of FeB2+ to FeC2+ enables the oxidation of three Fe2+ ions. The transient binding of Fe2+ at one of three newly observed Zn2+ sites may allow the oxidation of four Fe2+ by one dioxygen molecule. PMID- 12730191 TI - TTF-1, a homeodomain-containing transcription factor, participates in the control of body fluid homeostasis by regulating angiotensinogen gene transcription in the rat subfornical organ. AB - In recent years, it has become increasingly evident that angiotensins synthesized in the brain contribute to regulating body fluid homeostasis. Although angiotensinogen, the unique angiotensin precursor, is produced in the brain, the factors that regulate its gene expression remain unknown. We recently found that TTF-1, a homeodomain-containing transcription factor essential for the development of the fetal diencephalon, is postnatally expressed in discrete areas of the hypothalamus. We now report that the subfornical organ, an important site of angiotensinogen synthesis, is an extra-hypothalamic site of TTF-1 expression. Double in situ hybridization histochemistry demonstrated the presence of TTF-1 mRNA in angiotensinogen-producing cells of the rat subfornical organ. RNase protection assays showed that TTF-1 and angiotensinogen mRNA levels are simultaneously increased in the subfornical organ by water deprivation. The angiotensinogen promoter contains seven presumptive TTF-1 binding motifs, four of which are recognized by the TTF-1 homeodomain. In the C6 glioma cell line, TTF-1 transactivates the angiotensinogen promoter in a dose-dependent manner. This transactivation is abolished by deletion of the TTF-1 binding motif at -125. Intracranial administration of an antisense TTF-1 oligodeoxynucleotide decreased angiotensinogen mRNA in the subfornical organ and dramatically reduced the animal's water intake while increasing urine excretion. Moreover, plasma arginine vasopressin content was decreased by the same treatment. These results demonstrate a novel role for TTF-1 in the regulation of body fluid homeostasis, exerted via the transactivational control of angiotensinogen synthesis in the subfornical organ. PMID- 12730193 TI - The biology and enzymology of protein tyrosine O-sulfation. PMID- 12730192 TI - Presence of multiple mRNA cycling sequence element-binding proteins in Crithidia fasciculata. AB - A consensus sequence present in the 5'- or 3'-untranslated regions of several Crithidia fasciculata messenger RNAs encoding proteins involved in DNA metabolism has been shown to be necessary for the periodic accumulation of these mRNAs during the cell cycle. A protein complex termed cycling sequence-binding protein (CSBP) has two subunits, CSBPA and CSBPB, and binds the consensus sequence with high specificity. The binding activity of CSBP was shown to vary during the cell cycle in parallel with the levels of putative target mRNAs. Although disruption of the CSBPA gene resulted in loss of both CSBPA and CSBPB, the putative target message levels still continued to vary during the cell cycle. The presence of an additional and distinct binding activity was revealed in these CSBPA null mutant cells. This activity, termed CSBP II, was also expressed in wild-type Crithidia cells. CSBP II has higher binding specificity for the cycling sequence element than the earlier described CSBP complex. Three polypeptides associated with purified CSBP II show specific binding to the cycling sequence. These proteins may represent a family of sequence-specific RNA-binding proteins involved in post transcriptional regulation. PMID- 12730194 TI - A calmodulin/inositol 1,4,5-trisphosphate (IP3) receptor-binding region targets TRPC3 to the plasma membrane in a calmodulin/IP3 receptor-independent process. AB - Conformational coupling with the inositol 1,4,5-trisphosphate (IP3) receptor has been suggested as a possible mechanism of activation of TRPC3 channels and a region in the C terminus of TRPC3 has been shown to interact with the IP3 receptor as well as calmodulin (calmodulin/IP3 receptor-binding (CIRB) region). Here we show that internal deletion of 20 amino acids corresponding to the highly conserved CIRB region results in the loss of diacylglycerol and agonist-mediated channel activation in HEK293 cells. By using confocal microscopy to examine the cellular localization of Topaz fluorescent protein fusion constructs, we demonstrate that this loss in activity is caused by faulty targeting of CIRB deleted mutants to intracellular compartments. Wild type TRPC3 and mutants lacking a C-terminal predicted coiled coil region downstream of CIRB were targeted to the plasma membrane correctly in HEK293 cells and exhibited TRPC3 mediated calcium entry in response to agonist activation. Mutation of conserved YQ and MKR motifs to alanine within the CIRB region in TRPC3-Topaz, which would be expected to interfere with IP3 receptor and/or calmodulin binding, had no effect on channel function or targeting. Additionally, TRPC3 targets to the plasma membrane of DT40 cells lacking all three IP3 receptors and forms functional ion channels. These findings indicate that the previously identified CIRB region of TRPC3 is involved in its targeting to the plasma membrane by a mechanism that does not involve interaction with IP3 receptors. PMID- 12730196 TI - Design of potent peptide mimetics of brain-derived neurotrophic factor. AB - Brain-derived neurotrophic factor (BDNF) has potential for the treatment of human neurodegenerative diseases. However, the general lack of success of neurotrophic factors in clinical trials has led to the suggestion that low molecular weight neurotrophic drugs may be better agents for therapeutic use. Here we describe small, dimeric peptides designed to mimic a pair of solvent-exposed loops important for the binding and activation of the BDNF receptor, trkB. The monomer components that make up the dimers were based on a monocyclic monomeric peptide mimic of a single loop of BDNF (loop 2) that we had previously shown to be an inhibitor of BDNF-mediated neuronal survival (O'Leary, P. D., and Hughes, R. A. (1998) J. Neurochem. 70, 1712-1721). Bicyclic dimeric peptides behaved as partial agonists with respect to BDNF, promoting the survival of embryonic chick sensory neurons in culture. We reasoned that the potency and/or efficacy of these compounds might be improved by reducing the conformational flexibility about their dimerizing linker. Thus, we designed a highly conformationally constrained tricyclic dimeric peptide and synthesized it using an efficient, quasi-one-pot approach. Although still a partial BDNF-like agonist, the tricyclic dimer was particularly potent in promoting neuronal survival in vitro (EC50 11 pm). The peptides described here, which are greatly reduced in size compared with the parent protein, could serve as useful lead compounds for the development of true neurotrophic drugs and indicate that the structure-based design approach could be used to obtain potent mimetics of other growth factors that dimerize their receptors. PMID- 12730197 TI - Nuclear thiol peroxidase as a functional alkyl-hydroperoxide reductase necessary for stationary phase growth of Saccharomyces cerevisiae. AB - Yeast nucleus-localized thiol peroxidase (nTPx) was characterized as a functional peroxidase. There are two cysteine residues in nTPx. Replacement of Cys-106 or Cys-111 with serine resulted in a complete loss of thioredoxin-linked peroxidase activity. However, when their activities were measured in terms of the ability to inhibit oxidation of glutamine synthetase, C111S showed the same antioxidant activity as the wild type protein. SDS-PAGE gel analysis revealed that only C111S existed as the dimer form. In addition to the identification of Cys-106 as the primary catalytic site, these data suggest the formation of the intradisulfide bond as a part of the catalytic cycle between nTPx and thioredoxin. nTPx preferentially reduced alkyl-hydroperoxides rather than H2O2. Furthermore, a nTPx mutant strain showed higher sensitivity toward alkyl-hydroperoxide than hydrogen peroxide. Also, reduction of the viability of nTPx mutant strain against various oxidants supports an in vivo antioxidant role for nTPx. nTPx transcriptional activity was not significantly detectable in log phase yeast, but the activity was exponentially increased after the diauxic shift. The transcriptional activity was highly induced even in the log phase yeast grown in nonfermentable carbon source. Deletion of Tor1p, Ras1p, and Ras2p resulted in considerable induction when compared with their parent strains, demonstrating the activation of the transcription of nTPx gene at the diauxic shift. Transcription of nTPx gene was induced in response to oxidative stress. Viability of a stationary phase nTPx mutant was considerably reduced compared with the isogenic strain. Collectively, these data demonstrate that nTPx is a thiol peroxidase family acting as alkyl hydroperoxide reductase in the nucleus during post-diauxic growth. PMID- 12730195 TI - Identification and characterization of BCL-3-binding protein: implications for transcription and DNA repair or recombination. AB - A putative oncogene bcl-3 was originally identified and cloned at the breakpoint in the recurring chromosome translocation t(14;19) found in some cases of B cell chronic lymphocytic leukemia. Studies of bcl-3-deficient mice demonstrated a critical role for bcl-3 in the development of a normal immune response and the formation of germinal centers in secondary lymphoid organs. However, the molecular mechanism that underlies B cell leukemogenesis and the knockout mouse phenotype remains unclear. Here we have identified and characterized BCL-3 binding protein (B3BP) as a protein interacting specifically with the bcl-3 gene product (BCL-3) by a yeast two-hybrid screen. We found that B3BP associates with not only BCL-3 but also p300/CBP histone acetyltransferases. The N-terminal region of B3BP that contains the ATP-binding site is important for the interaction with BCL-3 and p300/CBP. Homology searches indicate that the ATP binding region of B3BP, which contains a typical Walker-type ATP-binding P-loop, most resembles that of 2',3'-cyclic nucleotide 3'-phosphodiesterase of mammals and polynucleotide kinase of T4 bacteriophage. In fact B3BP shows intrinsic ATP binding and hydrolyzing activity. Furthermore, we demonstrated that B3BP is a 5' polynucleotide kinase. We also found a small MutS-related domain, which is thought to be involved in the DNA repair or recombination reaction, in the C terminal region of B3BP, and it shows nicking endonuclease activity. These observations might help to gain new insights into the function of BCL-3 and p300/CBP, especially the coupling of transcription with repair or recombination. PMID- 12730198 TI - The ubiquinone-binding site in NADH:ubiquinone oxidoreductase from Escherichia coli. AB - An azido-ubiquinone derivative, 3-azido-2-methyl-5-methoxy[3H]-6-decyl-1,4 benzoquinone ([3H]azido-Q), was used to study the ubiquinone/protein interaction and to identify the ubiquinone-binding site in Escherichia coli NADH:ubiquinone oxidoreductase (complex I). The purified complex I showed no loss of activity after incubation with a 20-fold molar excess of [3H]azido-Q in the dark. Illumination of the incubated sample with long wavelength UV light for 10 min at 0 degrees C caused a 40% decrease of NADH:ubiquinone oxidoreductase activity. SDS PAGE of the complex labeled with [3H]azido-Q followed by analysis of the radioactivity distribution among the subunits revealed that subunit NuoM was heavily labeled, suggesting that this protein houses the Q-binding site. When the [3H]azido-Q-labeled NuoM was purified from the labeled reductase by means of preparative SDS-PAGE, a 3-azido-2-methyl-5-methoxy-6-decyl-1,4-benzoquinone linked peptide, with a retention time of 41.4 min, was obtained by high performance liquid chromatography of the protease K digest of the labeled subunit. This peptide had a partial NH2-terminal amino acid sequence of NH2 VMLIAILALV-, which corresponds to amino acid residues 184-193 of NuoM. The secondary structure prediction of NuoM using the Toppred hydropathy analysis showed that the Q-binding peptide overlaps with a proposed Q-binding motif located in the middle of the transmembrane helix 5 toward the cytoplasmic side of the membrane. Using the PHDhtm hydropathy plot, the labeled peptide is located in the transmembrane helix 4 toward the periplasmic side of the membrane. PMID- 12730199 TI - Role of the SCFSkp2 ubiquitin ligase in the degradation of p21Cip1 in S phase. AB - The cyclin-dependent kinase inhibitor p21Cip1 has important roles in the control of cell proliferation, differentiation, senescence, and apoptosis. It has been observed that p21 is a highly unstable protein, but the mechanisms of its degradation remained unknown. We show here that p21 is a good substrate for an SCF (Skp1-Cullin1-F-box protein) ubiquitin ligase complex, which contains the F box protein Skp2 (S phase kinase-associated protein 2) and the accessory protein Cks1 (cyclin kinase subunit 1). A similar ubiquitin ligase complex has been previously shown to be involved in the degradation of a related cyclin-dependent kinase inhibitor, p27Kip1. The levels of Skp2 oscillate in the cell cycle, reaching a maximum in S phase. The ubiquitylation of p21 in vitro required the supplementation of all components of the SCF complex as well as of Cks1 and Cdk2 cyclin E. The protein kinase Cdk2-cyclin E acts both by the phosphorylation of p21 on Ser-130 and by the formation of a complex with p21, which is required for its presentation to the ubiquitin ligase. As opposed to the case of p27, the phosphorylation of p21 stimulates its ubiquitylation but is not absolutely required for this process. Levels of p21 are higher in Skp2-/- mouse embryo fibroblasts than in wild-type fibroblasts in the S phase, and the rates of the degradation of p21 are slower in cells that lack Skp2. It is suggested that SCFSkp2 participates in the degradation of p21 in the S phase. PMID- 12730200 TI - Gene cluster of Arthrobacter ilicis Ru61a involved in the degradation of quinaldine to anthranilate: characterization and functional expression of the quinaldine 4-oxidase qoxLMS genes. AB - A genetic analysis of the anthranilate pathway of quinaldine degradation was performed. A 23-kb region of DNA from Arthrobacter ilicis Ru61a was cloned into the cosmid pVK100. Although Escherichia coli clones containing the recombinant cosmid did not transform quinaldine, cosmids harboring the 23-kb region, or a 10.8-kb stretch of this region, conferred to Pseudomonas putida KT2440 the ability to cometabolically convert quinaldine to anthranilate. The 10.8-kb fragment thus contains the genes coding for quinaldine 4-oxidase (Qox), 1H-4 oxoquinaldine 3-monooxygenase, 1H-3-hydroxy-4-oxoquinaldine 2,4-dioxygenase, and N-acetylanthranilate amidase. The qoxLMS genes coding for the molybdopterin cytosine dinucleotide-(MCD-), FeSI-, FeSII-, and FAD-containing Qox were inserted into the expression vector pJB653, generating pKP1. Qox is the first MCD containing enzyme to be synthesized in a catalytically fully competent form by a heterologous host, P. putida KT2440 pKP1; the catalytic properties and the UV visible and EPR spectra of Qox purified from P. putida KT2440 pKP1 were essentially like those of wild-type Qox. This provides a starting point for the construction of protein variants of Qox by site-directed mutagenesis. Downstream of the qoxLMS genes, a putative gene whose deduced amino acid sequence showed 37% similarity to the cofactor-inserting chaperone XdhC was located. Additional open reading frames identified on the 23-kb segment may encode further enzymes (a glutamyl tRNA synthetase, an esterase, two short-chain dehydrogenases/reductases, an ATPase belonging to the AAA family, a 2-hydroxyhepta-2,4-diene-1,7-dioate isomerase/5-oxopent-3-ene-1,2,5-tricarboxylate decarboxylase-like protein, and an enzyme of the mandelate racemase group) and hypothetical proteins involved in transcriptional regulation, and metabolite transport. PMID- 12730201 TI - Ca2+-dependent phosphorylation of syntaxin-1A by the death-associated protein (DAP) kinase regulates its interaction with Munc18. AB - Syntaxin-1 is a key component of the synaptic vesicle docking/fusion machinery that binds with VAMP/synaptobrevin and SNAP-25 to form the SNARE complex. Modulation of syntaxin binding properties by protein kinases could be critical to control of neurotransmitter release. Using yeast two-hybrid selection with syntaxin-1A as bait, we have isolated a cDNA encoding the C-terminal domain of death-associated protein (DAP) kinase, a calcium/calmodulin-dependent serine/threonine protein kinase. Expression of DAP kinase in adult rat brain is restricted to particular neuronal subpopulations, including the hippocampus and cerebral cortex. Biochemical studies demonstrate that DAP kinase binds to and phosphorylates syntaxin-1 at serine 188. This phosphorylation event occurs both in vitro and in vivo in a Ca2+-dependent manner. Syntaxin-1A phosphorylation by DAP kinase or its S188D mutant, which mimics a state of complete phosphorylation, significantly decreases syntaxin binding to Munc18-1, a syntaxin-binding protein that regulates SNARE complex formation and is required for synaptic vesicle docking. Our results suggest that syntaxin is a DAP kinase substrate and provide a novel signal transduction pathway by which syntaxin function could be regulated in response to intracellular [Ca2+] and synaptic activity. PMID- 12730202 TI - The two upstream open reading frames of oncogene mdm2 have different translational regulatory properties. AB - Few details are known of the mechanisms through which multiple upstream open reading frames (uORFs) interact to regulate translation in higher eukaryotes. The predominant transcript of oncogene mdm2 in normal human cells (L-mdm2) contains two upstream open reading frames in its 5' leader. Elimination of these two uORFs raises the translational efficiency of the transcript by over 10-fold in HeLa cells. The 5'-most uORF (uORF1) alone suppresses downstream translational activity by over 5-fold, whereas uORF2 contributes <2-fold to the inhibition by the intact leader. The different activities of the two uORFs do not depend on the nucleotide sequence surrounding the uORFs in the 5' leader, the order of the two uORFs in the 5' leader, or the occurrence of secondary structure or rare codons within the uORFs. Specific features of the amino acid sequence encoded by uORF1 contribute to its stronger suppressive activity, suggesting that it belongs to the class of "sequence-specific" uORFs. The weaker inhibitory activity inherent in uORF2 is potentiated by a sub-optimal nucleotide context surrounding its initiator AUG. The occurrence of two uORFs with differing activities in both the human gene and the mouse orthologue suggests that this pair of elements may play a fundamental role in regulating expression of the mdm2 gene. PMID- 12730203 TI - Expression and functional dynamics of the XCAP-D2 condensin subunit in Xenopus laevis oocytes. AB - The 13 S condensin complex plays a crucial role in the condensation and segregation of the two sets of chromosomes during mitosis in vivo as well as in cell-free extracts. This complex, conserved from yeast to human, contains a heterodimer of structural maintenance of chromosome (SMC) family proteins and three additional non-SMC subunits. We have investigated the expression of the non SMC condensin component XCAP-D2 in Xenopus laevis oocytes. When studied during meiotic maturation, XCAP-D2 starts to accumulate at the time of germinal vesicle breakdown and reaches its maximal amount in metaphase II oocytes. This accumulation is specifically blocked by injection of antisense oligonucleotides. XCAP-D2 antisense-injected oocytes progress normally through meiosis until metaphase II. At this stage, however, chromosomes exhibit architecture defaults, and resolution of sister chromatids is impaired. Surprisingly, in mitotic extracts made from XCAP-D2 knocked-down oocytes, sperm chromatin normally condenses into compacted chromosomes, whereas the amounts of both free and chromosome-bound XCAP-D2 are markedly reduced. This apparent discrepancy is discussed in light of current knowledge on chromosome dynamics. PMID- 12730205 TI - The Vid vesicle to vacuole trafficking event requires components of the SNARE membrane fusion machinery. AB - The key gluconeogenic enzyme fructose-1,6-bisphosphatase (FBPase) is targeted to Vid vesicles when glucose-starved cells are replenished with glucose. Vid vesicles then deliver FBPase to the vacuole for degradation. A modified alkaline phosphatase assay was developed to study the trafficking of Vid vesicles to the vacuole. For this assay, FBPase was fused with a truncated form of alkaline phosphatase. Under in vivo conditions, FBPase-delta60Pho8p was targeted to the vacuole via Vid vesicles, and it exhibited Pep4p- and Vid24p-dependent alkaline phosphatase activation. Vid vesicle-vacuole targeting was reconstituted using Vid vesicles that contained FBPase-delta60Pho8p. These vesicles were incubated with vacuoles in the presence of cytosol and an ATP-regenerating system. Under in vitro conditions, alkaline phosphatase was also activated in a Pep4p- and Vid24p dependent manner. The GTPase Ypt7p was identified as an essential component in Vid vesicle-vacuole trafficking. Likewise, a number of v-SNAREs (Ykt6p, Nyv1p, Vti1p) and homotypic fusion vacuole protein sorting complex family members (Vps39p and Vps41p) were required for the proper function of Vid vesicles. In contrast, the t-SNARE Vam3p was a necessary vacuolar component. Vid vesicle vacuole trafficking exhibits characteristics similar to heterotypic membrane fusion events. PMID- 12730204 TI - Overexpression of plasma membrane-associated sialidase attenuates insulin signaling in transgenic mice. AB - Plasma membrane-associated sialidase is a key enzyme for ganglioside hydrolysis, thereby playing crucial roles in regulation of cell surface functions. Here we demonstrate that mice overexpressing the human ortholog (NEU3) develop diabetic phenotype by 18-22 weeks associated with hyperinsulinemia, islet hyperplasia, and increased beta-cell mass. As compared with the wild type, insulin-stimulated phosphorylation of the insulin receptor (IR) and insulin receptor substrate I was significantly reduced, and activities of phosphatidylinositol 3-kinase and glycogen synthase were low in transgenic muscle. IR phosphorylation was already attenuated in the younger mice before manifestation of hyperglycemia. Transient transfection of NEU3 into 3T3-L1 adipocytes and L6 myocytes caused a significant decrease in IR signaling. In response to insulin, NEU3 was found to undergo tyrosine phosphorylation and subsequent association with the Grb2 protein, thus being activated and causing negative regulation of insulin signaling. In fact, accumulation of GM1 and GM2, the possible sialidase products in transgenic tissues, caused inhibition of IR phosphorylation in vitro, and blocking of association with Grb2 resulted in reversion of impaired insulin signaling in L6 cells. The data indicate that NEU3 indeed participates in the control of insulin signaling, probably via modulation of gangliosides and interaction with Grb2, and that the mice can serve as a valuable model for human insulin-resistant diabetes. PMID- 12730206 TI - Identification and characterization of a novel interaction between pulmonary surfactant protein D and decorin. AB - Surfactant-associated protein D (SP-D) is a collectin that is present in lung surfactant and mucosal surfaces. Although SP-D regulates diverse functions, only a few proteins are known to bind to this collectin. Here we describe the co purification of decorin, a novel SP-D-binding protein, from amniotic fluid. The human decorin that co-purified with SP-D is a 130-150-kDa proteoglycan, which has a 46-kDa protein core and approximately 90-kDa dermatan sulfate chain. Both native and recombinant decorin can bind to SP-D that is already bound to maltose agarose matrix, and these SP-D-decorin complexes are dissociated at high salt (0.5-1.0 m NaCl) conditions, releasing the decorin. We further show that SP-D and decorin interact with each other (kd = 4 nm) by two mechanisms. First, the direct binding and competition experiments show that the carbohydrate recognition domain (CRD) of SP-D binds in a calcium dependent-manner to the sulfated N-acetyl galactosamine moiety of the glycosaminoglycan chain. Second, complement component C1q, a complement protein that is known to interact with decorin core protein via its collagen-like region, partially blocks the interaction between decorin and native SP-D. This protein, however, does not block the interaction between decorin and SP-D(n/CRD), a recombinant fragment that lacks the N-terminal and collagen-like regions. Furthermore, the core protein, obtained by chondroitin ABC lyase treatment of decorin, binds SP-D, but not SP-D(n/CRD). These findings suggest that decorin core protein binds the collagen-like region of the SP-D. Concentrations of decorin and SP-D are negatively correlated to each other, in amniotic fluid, implying a functional relevance for SP-D-decorin interaction, in vivo. Collectively, our results show that carbohydrate recognition domains of SP D interact with the dermatan sulfate moiety of decorin via lectin activity and that the core protein of decorin binds the collagen-like region of SP-D in vitro, and these interactions may be operative in vivo. PMID- 12730207 TI - Sigma 1- and mu 1-Adaptin homologues of Leishmania mexicana are required for parasite survival in the infected host. AB - The sorting of membrane-bound proteins from the trans-Golgi network to lysosomal/endosomal compartments is achieved by preferential inclusion into clathrin-coated vesicles. Contained within the cytoplasmic domains of such proteins, specific sequence motifs have been identified (tyrosine-based and/or di leucine-based) that are essential for targeting and are recognized by a family of heterotetrameric adaptor complexes, which then recruit clathrin. These cytosolic protein complexes, which have been found in a wide variety of higher eukaryotic organisms, are essential for the development of multicellular organisms. In trypanosomatids, the adaptin-mediated sorting of proteins is largely uncharacterized. In order to identify components of the adaptor-complex machinery, this study reports the cloning and characterization of sigma 1- and mu 1-adaptin gene homologues from the eukaryotic protozoan parasite, Leishmania mexicana. Generation of sigma 1- and mu 1-adaptin gene deletion mutants shows that these promastigote parasites are viable in culture, but are unable to establish infection of macrophages or mice, indicating that adaptin function is crucial for pathogenesis in these unicellular organisms. PMID- 12730208 TI - Separation of lipid transport functions by mutations in the extracellular domain of scavenger receptor class B, type I. AB - Scavenger receptor class B, type I (SR-BI) shows a variety of effects on cellular cholesterol metabolism, including increased selective uptake of high density lipoprotein (HDL) cholesteryl ester, stimulation of free cholesterol (FC) efflux from cells to HDL and phospholipid vesicles, and changes in the distribution of plasma membrane FC as evidenced by increased susceptibility to exogenous cholesterol oxidase. Previous studies showed that these multiple effects require the extracellular domain of SR-BI, but not the transmembrane and cytoplasmic domains. To test whether 1) the extracellular domain of SR-BI mediates multiple activities by virtue of discrete functional subdomains, or 2) the multiple activities are, in fact, secondary to and driven by changes in cholesterol flux, the extracellular domain of SR-BI was subjected to insertional mutagenesis by strategically placing an epitope tag into nine sites. These experiments identified four classes of mutants with disruptions at different levels of function. Class 4 mutants showed a clear separation of function between HDL binding, HDL cholesteryl ester uptake, and HDL-dependent FC efflux on one hand and FC efflux to small unilamellar vesicles and an increased cholesterol oxidase sensitive pool of membrane FC on the other. Selective disruption of the latter two functions provides evidence for multiple functional subdomains in the extracellular receptor domain. Furthermore, these findings uncover a difference in the SR-BI-mediated efflux pathways for FC transfer to HDL acceptors versus phospholipid vesicles. The loss of the cholesterol oxidase-sensitive FC pool and FC efflux to small unilamellar vesicle acceptors in Class 4 mutants suggests that these activities may be mechanistically related. PMID- 12730210 TI - Structure of a Sir2 substrate, Alba, reveals a mechanism for deacetylation induced enhancement of DNA binding. AB - The targeted acetylation status of histones and several other transcriptional regulatory proteins plays an important role in gene expression, although the mechanism for this is not well understood. As a model to understand how targeted acetylation may effect transcription, we determined the x-ray crystal structure of the chromatin protein Alba from Archaeoglobus fulgidus, a substrate for the Sir2 protein that deacetylates it at lysine 11 to promote DNA binding by Alba. The structure reveals a dimer of dimers in which the dimer-dimer interface is stabilized by several conserved hydrophobic residues as well as the lysine 11 target of Sir2. We show that, in solution, the mutation of these hydrophobic residues or lysine 11 disrupts dimer-dimer formation and decreases DNA-binding affinity. We propose that the in vivo deacetylation of lysine 11 of archaeal Alba by Sir2 promotes protein oligomerization for optimal DNA binding. Implications for the mechanism by which histone acetylation modulates gene expression are discussed. PMID- 12730209 TI - Neurofibromatosis type I tumor suppressor neurofibromin regulates neuronal differentiation via its GTPase-activating protein function toward Ras. AB - Neurofibromin, the neurofibromatosis type 1 (NF1) gene product, contains a central domain homologous to a family of proteins known as Ras-GTPase-activating proteins (Ras-GAPs), which function as negative regulators of Ras. The loss of neurofibromin function has been thought to be implicated in the abnormal regulation of Ras in NF1-related pathogenesis. In this study, we found a novel role of neurofibromin in neuronal differentiation in conjunction with the regulation of Ras activity via its GAP-related domain (GRD) in neuronal cells. In PC12 cells, time-dependent increases in the GAP activity of cellular neurofibromin (NF1-GAP) were detected after NGF stimulation, which were correlated with the down-regulation of Ras activity during neurite elongation. Interestingly, the NF1-GAP increase was due to the induction of alternative splicing of NF1-GRD type I triggered by the NGF-induced Ras activation. Dominant negative (DN) forms of NF1-GRD type I significantly inhibited the neurite extension of PC12 cells via regulation of the Ras state. NF1-GRD-DN also reduced axonal and dendritic branching/extension of rat embryonic hippocampal neurons. These results demonstrate that the mutual regulation of Ras and NF1-GAP is essential for normal neuronal differentiation and that abnormal regulation in neuronal cells may be implicated in NF1-related learning and memory disturbance. PMID- 12730211 TI - Selective loss of trans-acting instability determinants of neurofilament mRNA in amyotrophic lateral sclerosis spinal cord. AB - Neurofilament (NF) aggregates in motor neurons are a key neuropathological feature of amyotrophic lateral sclerosis (ALS). We have previously observed an alteration in the stoichiometry of NF subunit steady state mRNA levels in ALS spinal motor neurons using in situ hybridization and proposed that this led to aggregate formation. We have now examined the levels of NF mRNA in whole tissue homogenates of spinal cord using the RNase protection assay and real time reverse transcriptase-PCR and observed significant elevations of NF mRNA level in ALS. Compared with age-matched control, we observed a greater stability of heterogeneously expressed NFL mRNA in the presence of ALS spinal cord homogenates. Heat denaturing or protease K digestion of the control homogenates increased the stability of the NFL mRNA to levels observed in ALS homogenate. Increased NFL mRNA stability was also induced by increasing the percentage of ALS homogenate in an admixture of control and ALS homogenates. These observations suggest the presence of trans-acting NFL mRNA-destabilizing elements in control but not in ALS spinal cord homogenates. This was confirmed in gel retardation assays. We also observed that the destabilizing elements interact with the 3' untranslated region of NFL mRNA. These findings suggest that the trans-acting NFL destabilizing elements are selectively suppressed in ALS homogenates, resulting in an increased stability and level of NFL mRNA. PMID- 12730212 TI - Depolymerization of actin filaments by profilin. Effects of profilin on capping protein function. AB - Profilin interacts with the barbed ends of actin filaments and is thought to facilitate in vivo actin polymerization. This conclusion is based primarily on in vitro kinetic experiments using relatively low concentrations of profilin (1-5 microm). However, the cell contains actin regulatory proteins with multiple profilin binding sites that potentially can attract millimolar concentrations of profilin to areas requiring rapid actin filament turnover. We have studied the effects of higher concentrations of profilin (10-100 microm) on actin monomer kinetics at the barbed end. Prior work indicated that profilin might augment actin filament depolymerization in this range of profilin concentration. At barbed-end saturating concentrations (final concentration, approximately 40 microm), profilin accelerated the off-rate of actin monomers by a factor of four to six. Comparable concentrations of latrunculin had no detectable effect on the depolymerization rate, indicating that profilin-mediated acceleration was independent of monomer sequestration. Furthermore, we have found that high concentrations of profilin can successfully compete with CapG for the barbed end and uncap actin filaments, and a simple equilibrium model of competitive binding could explain these effects. In contrast, neither gelsolin nor CapZ could be dissociated from actin filaments under the same conditions. These differences in the ability of profilin to dissociate capping proteins may explain earlier in vivo data showing selective depolymerization of actin filaments after microinjection of profilin. The finding that profilin can uncap actin filaments was not previously appreciated, and this newly discovered function may have important implications for filament elongation as well as depolymerization. PMID- 12730213 TI - Disruption of basal JNK activity differentially affects key fibroblast functions important for wound healing. AB - We used both a gene knockout approach and pharmacologic modulation to study the implication of the JNK pathway in regulating fibroblast motility, capacity to contract mechanically unloaded collagen gels, and type I collagen gene expression in vitro. These parameters, which are important for tissue repair, are positively regulated by transforming growth factor (TGF)-beta, a cytokine viewed as playing a master role during wound healing. We demonstrate that basal JNK activity is critical for fibroblast motility because (a) mouse embryo jnk-/- fibroblasts exhibit significantly lower ability to close mechanically induced cell layer wounds than their wild-type (wt) counterparts, and (b) wound closure by human dermal fibroblasts is dramatically impaired by the specific JNK inhibitor SP600125. junAA fibroblasts, in which amino acids Ser63 and Ser73 of c-Jun are replaced by two Ala residues so that c-Jun cannot be phosphorylated by JNK, also exhibited impaired motility, suggesting that c-Jun phosphorylation by JNK is critical for fibroblast migration. In sharp contrast to their lesser motility on plastic, jnk-/- and junAA fibroblasts contracted free-floating, mechanically unloaded, collagen lattices markedly faster than wt fibroblasts. Furthermore, basal mRNA steady-state levels for types I and III collagen genes were similar in jnk-/- and wt fibroblasts. Likewise, overexpression of a dominant-negative mutant form of MKK4 in dermal fibroblasts did not affect collagen expression. We also demonstrate that basal JNK activity does not affect either TGF-beta-induced collagen gene expression or lattice contraction, whereas on the other hand, the blockage of motility initiated by JNK inhibition cannot be overcome by TGF-beta. Together these results demonstrate discrete, yet significant and highly specific, regulation of fibroblast functions important for wound healing by basal JNK activity. PMID- 12730214 TI - Anionic micelles and vesicles induce tau fibrillization in vitro. AB - Alzheimer's disease is defined in part by the intraneuronal accumulation of filaments comprised of the microtubule-associated protein tau. In vitro, fibrillization of recombinant tau can be induced by treatment with various agents, including phosphotransferases, polyanionic compounds, and fatty acids. Here we characterize the structural features required for the fatty acid class of tau fibrillization inducer using recombinant full-length tau protein, arachidonic acid, and a series of straight chain anionic, cationic, and nonionic detergents. Induction of measurable tau fibrillization required an alkyl chain length of at least 12 carbons and a negative charge consisting of carboxylate, sulfonate, or sulfate moieties. All detergents and fatty acids were micellar at active concentrations, due to a profound, taudependent depression of their critical micelle concentrations. Anionic surfaces larger than detergent micelles, such as those supplied by phosphatidylserine vesicles, also induced tau fibrillization with resultant filaments originating from their surface. These data suggest that anionic surfaces presented as micelles or vesicles can serve to nucleate tau fibrillization, that this mechanism underlies the activity of fatty acid inducers, and that anionic membranes may serve this function in vivo. PMID- 12730215 TI - Chimeric enzymes of cytochrome P450 oxidoreductase and neuronal nitric-oxide synthase reductase domain reveal structural and functional differences. AB - The nitric-oxide synthases (NOSs) are comprised of an oxygenase domain and a reductase domain bisected by a calmodulin (CaM) binding region. The NOS reductase domains share approximately 60% sequence similarity with the cytochrome P450 oxidoreductase (CYPOR), which transfers electrons to microsomal cytochromes P450. The crystal structure of the neuronal NOS (nNOS) connecting/FAD binding subdomains reveals that the structure of the nNOS-connecting subdomain diverges from that of CYPOR, implying different alignments of the flavins in the two enzymes. We created a series of chimeric enzymes between nNOS and CYPOR in which the FMN binding and the connecting/FAD binding subdomains are swapped. A chimera consisting of the nNOS heme domain and FMN binding subdomain and the CYPOR FAD binding subdomain catalyzed significantly increased rates of cytochrome c reduction in the absence of CaM and of NO synthesis in its presence. Cytochrome c reduction by this chimera was inhibited by CaM. Other chimeras consisting of the nNOS heme domain, the CYPOR FMN binding subdomain, and the nNOS FAD binding subdomain with or without the tail region also catalyzed cytochrome c reduction, were not modulated by CaM, and could not transfer electrons into the heme domain. A chimera consisting of the heme domain of nNOS and the reductase domain of CYPOR reduced cytochrome c and ferricyanide at rates 2-fold higher than that of native CYPOR, suggesting that the presence of the heme domain affected electron transfer through the reductase domain. These data demonstrate that the FMN subdomain of CYPOR cannot effectively substitute for that of nNOS, whereas the FAD subdomains are interchangeable. The differences among these chimeras most likely result from alterations in the alignment of the flavins within each enzyme construct. PMID- 12730218 TI - Forcing chromatin. PMID- 12730216 TI - Neurotoxic mechanisms caused by the Alzheimer's disease-linked Swedish amyloid precursor protein mutation: oxidative stress, caspases, and the JNK pathway. AB - Autosomal dominant forms of familial Alzheimer's disease (FAD) are caused by mutations of the amyloid precursor protein (APP) gene and by mutations of the genes encoding for presenilin 1 or presenilin 2. Simultaneously, evidence is provided that increased oxidative stress might play a crucial role in the rapid progression of the Swedish FAD. Here we investigated the effect of the Swedish double mutation (K670M/N671L) in the beta-amyloid precursor protein on oxidative stress-induced cell death mechanisms in PC12 cells. Western blot analysis and cleavage studies of caspase substrates revealed an elevated activity of the executor caspase 3 after treatment with hydrogen peroxide in cells containing the Swedish APP mutation. This elevated activity is the result of the enhanced activation of both intrinsic and extrinsic apoptosis pathways, including activation of caspase 2 and caspase 8. Furthermore, we observed an enhanced activation of JNK pathway and an attenuation of apoptosis by SP600125, a JNK inhibitor, through protection of mitochondrial dysfunction and reduction of caspase 9 activity. Our findings provide evidence that the massive neurodegeneration in early age of FAD patients could be a result of an increased vulnerability of neurons through activation of different apoptotic pathways as a consequence of elevated levels of oxidative stress. PMID- 12730219 TI - Properties of the mouse intestinal acyl-CoA:monoacylglycerol acyltransferase, MGAT2. AB - Acyl-CoA:monoacylglycerol acyltransferase (MGAT) plays an important role in dietary fat absorption by catalyzing a rate-limiting step in the re-synthesis of diacylglycerols in enterocytes. The present study reports further characterization of MGAT2, a newly identified intestinal MGAT (Cao, J., Lockwood, J., Burn, P., and Shi, Y. (2003) J. Biol. Chem. 278, 13860-13866) for its substrate specificity, requirement for lipid cofactors, optimum pH and Mg2+, and other intrinsic properties. MGAT2 enzyme expressed in COS-7 cells displayed a broad range of substrate specificity toward fatty acyl-CoA derivatives and monoacylglycerols, among which the highest activities were observed with oleoyl CoA and rac-1-monolauroylglycerol, respectively. MGAT2 appeared to acylate monoacylglycerols containing unsaturated fatty acyls in preference to saturated ones. Lipid cofactors that play roles in signal transduction were shown to modulate MGAT2 activities. In contrast to oleic acid and sphingosine that exhibited inhibitory effects, phosphatidylcholine, phosphatidylserine, and phosphatidic acid stimulated MGAT2 activities. Using recombinant murine MGAT2 expressed in Escherichia coli, we demonstrated conclusively that MGAT2 also possessed an intrinsic acyl-CoA:diacylglycerol acyltransferase (DGAT) activity, which could provide an alternative pathway for triacylglycerol synthesis in the absence of DGAT. In contrast to the inhibitory effect on MGAT2 activities, nonionic and zwitterionic detergents led to a striking activation of DGAT activity of the human DGAT1 expressed in mammalian cells, which further distinguished the behaviors of the two enzymes. The elucidation of properties of MGAT2 will facilitate future development of compounds that inhibit dietary fat absorption as a means to treat obesity. PMID- 12730217 TI - Rho-dependent Rho kinase activation increases CD44 surface expression and bone resorption in osteoclasts. AB - Osteoclasts from osteopontin-deficient mice exhibit decreased CD44 surface expression [corrected]. Osteopontin (OPN)/alphavbeta3 generated Rho signaling pathway is required for the surface expression of CD44. In this work we show the Rho effector, Rho kinase (ROK-alpha), to be a potent activator of CD44 surface expression. ROK-alpha activation was associated with autophosphorylation, leading to its translocation to the plasma membrane, as well as its association with CD44. ROK-alpha promoted CD44 surface expression through phosphorylation of CD44 and ezrin-radixin-moesin (ERM) proteins and CD44.ERM.actin complex formation. Osteoclasts from OPN-/- mice exhibited an approximately 55-60% decrease in basal level ROK-alpha phosphorylation as compared with wild type osteoclasts. Furthermore, RhoVal-14 transduction was only partially effective in stimulating ROK-alpha/CD44 phosphorylation, as well as CD44 surface expression, in these osteoclasts. Studies on the inhibition of Rho by C3 transferase or ROK-alpha by the specific inhibitor, Y-27632, showed a decrease in the phosphorylation mediated by ROK-alpha and CD44 surface expression. Neutralizing antibodies to alphav, beta3, or CD44 inhibited the migration and bone resorption of wild type osteoclasts. However, only anti-alphav or -beta3 antibodies blocked OPN-induced phosphorylation of ROK-alpha, CD44, and the ERM proteins. Our results strongly suggest a role for ROK-alpha in alphavbeta3-mediated Rho signaling, which is required for the phosphorylation events and CD44 surface expression. The functional deficiencies in the Rho effector(s) because of the lack of OPN were associated with decreased CD44 surface expression and hypomotility in the OPN-/- osteoclasts. Finally, we find that cooperativity exists between alphavbeta3 and CD44 for osteoclast motility and bone resorption. PMID- 12730221 TI - NEDP1, a highly conserved cysteine protease that deNEDDylates Cullins. AB - The ubiquitin-like protein NEDD8 is essential for activity of SCF-like ubiquitin ligase complexes. Here we identify and characterize NEDP1, a human NEDD8-specific protease. NEDP1 is highly conserved throughout evolution and equivalent proteins are present in yeast, plants, insects, and mammals. Bacterially expressed NEDP1 is capable of processing NEDD8 in vitro to expose the diglycine motif required for conjugation and can deconjugate NEDD8 from modified substrates. NEDP1 appears to be specific for NEDD8 as neither ubiquitin nor SUMO bearing COOH-terminal extensions are utilized as substrates. Inhibition studies and mutagenesis indicate that NEDP1 is a cysteine protease with sequence similarities to SUMO specific proteases and the class of viral proteases typified by the adenovirus protease. In vivo NEDP1 deconjugates NEDD8 from a wide variety of substrates including the cullin component of SCF-like complexes. Thus NEDP1 is likely to play an important role in ubiquitin-mediated proteolysis by controlling the activity of SCF complexes. PMID- 12730220 TI - Nuclear accumulation of the small GTPase Gsp1p depends on nucleoporins Nup133p, Rat2p/Nup120p, Nup85p, Nic96p, and the acetyl-CoA carboxylase Acc1p. AB - The small GTPase Ran/Gsp1p plays an essential role in nuclear trafficking of macromolecules, as Ran/Gsp1p regulates many transport processes across the nuclear pore complex (NPC). To determine the role of nucleoporins in the generation of the nucleocytoplasmic Gsp1p concentration gradient, mutations in various nucleoporin genes were analyzed in the yeast Saccharomyces cerevisiae. We show that the nucleoporins Nup133p, Rat2p/Nup120p, Nup85p, Nic96p, and the enzyme acetyl-CoA carboxylase (MTR7) control the distribution and cellular concentration of Gsp1p. At the restrictive temperature the reporter protein GFP-Gsp1p, which is too large to diffuse across the nuclear envelope, fails to concentrate in nuclei of nup133delta, rat2-1, nup85delta, nic96deltaC, and mtr7-1 cells, demonstrating that GFP-Gsp1p nuclear import is deficient. In addition, the concentration of Gsp1p is severely reduced in mutants nup133Delta and mtr7-1 under these conditions. We have now identified the molecular mechanisms that contribute to the dissipation of the Gsp1p concentration gradient in these mutants. Loss of the Gsp1p gradient in nup133delta and rat2-1 can be explained by reduced binding of the Gsp1p nuclear carrier Ntf2p to NPCs. Likewise, nup85delta cells that mislocalize GFP-Gsp1p at the permissive as well as non-permissive temperature have a diminished association of Ntf2p-GFP with nuclear envelopes under both conditions. Moreover, under restrictive conditions Prp20p, the guanine nucleotide exchange factor for Gsp1p, mislocalizes to the cytoplasm in nup85delta, nic96deltaC, and mtr7-1 cells, thereby contributing to a collapse of the Gsp1p gradient. Taken together, components of the NPC subcomplex containing Rat2p/Nup120p, Nup133p, and Nup85p, in addition to proteins Nic96p and Mtr7p, are shown to be crucial for the formation of a nucleocytoplasmic Gsp1p gradient. PMID- 12730222 TI - UVB light stimulates production of reactive oxygen species: unexpected role for catalase. AB - In keratinocytes, UVB light stimulates the production of reactive oxygen species (ROS). Lysates of these cells were found to possess a non-dialyzable, trypsin- and heat-sensitive material capable of generating ROS in response to UVB light. Using ion exchange, metal affinity, and size exclusion chromatography, a 240-kDa protein was isolated with ROS generating activity. The protein exhibited strong absorption in the 320-360 nm range with additional soret peaks around 400-410 nm, suggesting the presence of heme. Sequencing using liquid chromatography-ion trap mass spectrometry identified the protein as catalase. Using purified catalases from a variety of species, the ROS generating activity was found to be temperature- and O2-dependent, stimulated by inhibitors of the catalatic activity of catalase, including 3-aminotriazole and azide, and inhibited by cyanide. A marked increase in the production of ROS was observed in UVB-treated cells overexpressing catalase and decreased generation of oxidants was found in UVB treated keratinocytes with reduced levels of catalase. Our data indicate that catalase plays a direct role in generating oxidants in response to UVB light. The finding that catalase mediates the production of ROS following UVB treatment is both novel and highly divergent from the well known antioxidant functions of the enzyme. We hypothesize that, through the actions of catalase, high energy DNA damaging UVB light is absorbed by the enzyme and converted to reactive chemical intermediates that can be detoxified by cellular antioxidant enzymes. Accumulation of excessive ROS, generated through the action of catalase, may lead to oxidative stress, DNA damage, and the development of skin cancer. PMID- 12730224 TI - Inhibition of human papillomavirus DNA replication by small molecule antagonists of the E1-E2 protein interaction. AB - Human papillomavirus (HPV) DNA replication is initiated by recruitment of the E1 helicase by the E2 protein to the viral origin. Screening of our corporate compound collection with an assay measuring the cooperative binding of E1 and E2 to the origin identified a class of small molecule inhibitors of the protein interaction between E1 and E2. Isothermal titration calorimetry and changes in protein fluorescence showed that the inhibitors bind to the transactivation domain of E2, the region that interacts with E1. These compounds inhibit E2 of the low risk HPV types 6 and 11 but not those of high risk HPV types or of cottontail rabbit papillomavirus. Functional evidence that the transactivation domain is the target of inhibition was obtained by swapping this domain between a sensitive (HPV11) and a resistant (cottontail rabbit papillomavirus) E2 type and by identifying an amino acid substitution, E100A, that increases inhibition by approximately 10-fold. This class of inhibitors was found to antagonize specifically the E1-E2 interaction in vivo and to inhibit HPV DNA replication in transiently transfected cells. These results highlight the potential of the E1-E2 interaction as a small molecule antiviral target. PMID- 12730223 TI - Fibronectin fragment activation of proline-rich tyrosine kinase PYK2 mediates integrin signals regulating collagenase-3 expression by human chondrocytes through a protein kinase C-dependent pathway. AB - Fibronectin fragments (FN-f), including the 110-kDa fragment that binds the alpha5beta1 integrin, stimulate collagenase-3 (MMP-13) production and cartilage destruction. In the present study, treatment of chondrocytes with the 110-kDa FN f or an activating antibody to the alpha5beta1 integrin was found to increase tyrosine autophosphorylation (Tyr-402) of the proline-rich tyrosine kinase-2 (PYK2) without significant change in autophosphorylation (Tyr-397) of focal adhesion kinase (FAK). The tyrosine kinase inhibitor tyrphostin A9, shown previously to block a PYK2-dependent pathway, blocked the FN-f-stimulated increase in MMP-13, whereas tyrphostin A25 did not. FN-f-stimulated PYK2 phosphorylation and MMP-13 production was also blocked by reducing intracellular calcium levels. Adenovirally mediated overexpression of wild type but not mutant PYK2 resulted in increased MMP-13 production. The protein kinase C (PKC) activator phorbol 12-myristate 13-acetate stimulated PYK2 phosphorylation and MMP 13 production. MMP-13 expression stimulated by either phorbol 12-myristate 13 acetate or FN-f was blocked by PKC inhibitors including the PKCdelta inhibitor rottlerin. Furthermore, PKCdelta translocation from cytosol to membrane was noted within 5 min of stimulation with FN-f. Immortalized human chondrocytes, transiently transfected with MMP-13 promoter-luciferase reporter constructs, showed increased promoter activity after FN-f treatment that was inhibited by co transfection with either of two dominant negative mutants of PYK2 (Y402F and K457A). No inhibition was seen after cotransfection with wild type PYK2, a dominant negative of FAK (FRNK) or empty vector plasmid. FN-f-stimulated MMP-13 promoter activity was also inhibited by chemical inhibitors of ERK, JNK, and p38 mitogen-activated protein (MAP) kinases or by co-transfection of dominant negative MAP kinase mutant constructs. These studies have identified a novel pathway for the MAP kinase regulation of MMP-13 production which involves FN-f stimulation of the alpha5beta1 integrin and activation of the nonreceptor tyrosine kinase PYK2 by PKC, most likely PKCdelta PMID- 12730225 TI - Cyclic AMP-dependent phosphorylation of cyclin D3-bound CDK4 determines the passage through the cell cycle restriction point in thyroid epithelial cells. AB - According to current concepts, the cell cycle commitment after restriction (R) point passage requires the sustained stimulation by mitogens of the synthesis of labile d-type cyclins, which associate with cyclin-dependent kinase (CDK) 4/6 to phosphorylate pRb family proteins and sequester the CDK inhibitor p27kip1. In primary cultures of dog thyroid epithelial cells, the cAMP-dependent cell cycle induced by a sustained stimulation by thyrotropin or forskolin differs from growth factor mitogenic pathways, as cAMP does not upregulate d-type cyclins but increases p27 levels. Instead, cAMP induces the assembly of required cyclin D3 CDK4 complexes, which associate with nuclear p27. In this study, the arrest of forskolin stimulation rapidly slowed down the entry of dog thyrocytes into S phase and the phosphorylation of pRb family proteins. The pRb kinase activity, but not the formation, of the cyclin D3-CDK4-p27 complex was strongly reduced. Using two-dimensional gel electrophoresis, a phosphorylated form of CDK4 was separated. It appeared in response to forskolin and was bound to both cyclin D3 and p27, presumably reflecting the activating Thr-172 phosphorylation of CDK4. Upon forskolin withdrawal or after cycloheximide addition, this CDK4 phosphoform unexpectedly persisted in p27 complexes devoid of cyclin D3 but it disappeared from the more labile cyclin D3 complexes. These data demonstrate that the assembly of the cyclin D3-CDK4-p27 holoenzyme and the subsequent phosphorylation and activation of CDK4 depend on distinct cAMP actions. This provides a first example of a crucial regulation of CDK4 phosphorylation by a mitogenic cascade and a novel mechanism of cell cycle control at the R point. PMID- 12730226 TI - Enzymology of base excision repair in the hyperthermophilic archaeon Pyrobaculum aerophilum. AB - DNA of all living organisms is constantly modified by exogenous and endogenous reagents. The mutagenic threat of modifications such as methylation, oxidation, and hydrolytic deamination of DNA bases is counteracted by base excision repair (BER). This process is initiated by the action of one of several DNA glycosylases, which removes the aberrant base and thus initiates a cascade of events that involves scission of the DNA backbone, removal of the baseless sugar phosphate residue, filling in of the resulting single nucleotide gap, and ligation of the remaining nick. We were interested to find out how the BER process functions in hyperthermophiles, organisms growing at temperatures around 100 degrees C, where the rates of these spontaneous reactions are greatly accelerated. In our previous studies, we could show that the crenarchaeon Pyrobaculum aerophilum has at least three uracil-DNA glycosylases, Pa-UDGa, Pa UDGb, and Pa-MIG, that can initiate the BER process by catalyzing the removal of uracil residues arising through the spontaneous deamination of cytosines. We now report that the genome of P. aerophilum encodes also the remaining functions necessary for BER and show that a system consisting of four P. aerophilum encoded enzymes, Pa-UDGb, AP endonuclease IV, DNA polymerase B2, and DNA ligase, can efficiently repair a G.U mispair in an oligonucleotide substrate to a G.C pair. Interestingly, the efficiency of the in vitro repair reaction was stimulated by Pa-PCNA1, the processivity clamp of DNA polymerases. PMID- 12730227 TI - Nonpolar thymine isosteres in the Ty3 polypurine tract DNA template modulate processing and provide a model for its recognition by Ty3 reverse transcriptase. AB - Despite diverging in sequence and size, the polypurine tract (PPT) primers of retroviruses and long terminal repeat-containing retrotransposons are accurately processed from (+) U3 RNA and DNA by their cognate reverse transcriptases (RTs). In this paper, we demonstrate that misalignment of the Ty3 retrotransposon RT on the human immunodeficiency virus-1 PPT induces imprecise removal of adjacent (+) RNA and failure to release (+)-DNA from the primer. Based on these observations, we explored the structural basis of Ty3 PPT recognition by chemically synthesizing RNA/DNA hybrids whose (-)-DNA template was substituted with the non hydrogen-bonding thymine isostere 2,4-difluoro-5-methylbenzene (F). We observed a consistent spatial correlation between the site of T --> F substitution and enhanced ribonuclease H (RNase H) activity approximately 12-13 bp downstream. In the most pronounced case, dual T --> F substitution at PPT positions -1/-2 redirects RNase H cleavage almost exclusively to the novel site. The structural features of this unusual base suggest that its insertion into the Ty3 PPT (-)-DNA template weakens the duplex, inducing a destabilization that is recognized by a structural element of Ty3 RT approximately 12-13 bp from its RNase H catalytic center. A likely candidate for this interaction is the thumb subdomain, whose minor groove binding tract most likely contacts the duplex. The spatial relationship derived from T --> F substitution also infers that Ty3 PPT processing requires recognition of sequences in its immediate 5' vicinity, thereby locating the RNase H catalytic center over the PPT-U3 junction, a notion strengthened by additional mutagenesis studies of this paper. PMID- 12730228 TI - Defining the SNARE complex binding surface of alpha-SNAP: implications for SNARE complex disassembly. AB - N-Ethylmaleimide-sensitive factor (NSF) and its adaptor protein alpha-soluble NSF attachment protein (alpha-SNAP) sustain membrane trafficking by disassembling soluble NSF attachment protein receptor (SNARE) complexes that form during membrane fusion. To better understand the role of alpha-SNAP in this process, we used site-directed mutagenesis to identify residues in alpha-SNAP that interact with SNARE complexes. We find that mutations in charged residues distributed over a concave surface formed by the N-terminal nine alpha-helices of alpha-SNAP affect its ability to bind synaptic SNARE complex and promote its disassembly by NSF. Replacing basic residues on this surface with alanines reduced SNARE complex binding and disassembly, whereas replacing acidic residues with alanines enhanced alpha-SNAP efficacy in both assays. These findings show that the ability of NSF to take apart SNARE complexes depends upon electrostatic interactions between alpha-SNAP and the acidic surface of the SNARE complex and provide insight into how NSF and alpha-SNAP work together to drive disassembly. PMID- 12730230 TI - The GTPase activity and C-terminal cysteine of the Escherichia coli MnmE protein are essential for its tRNA modifying function. AB - The Escherichia coli MnmE protein is a three-domain protein that exhibits a very high intrinsic GTPase activity and low affinity for GTP and GDP. The middle GTPase domain, when isolated, conserves the high intrinsic GTPase activity of the entire protein, and the C-terminal domain contains the only cysteine residue present in the molecule. MnmE is an evolutionarily conserved protein that, in E. coli, has been shown to control the modification of the uridine at the wobble position of certain tRNAs. Here we examine the biochemical and functional consequences of altering amino acid residues within conserved motifs of the GTPase and C-terminal domains of MnmE. Our results indicate that both domains are essential for the MnmE tRNA modifying function, which requires effective hydrolysis of GTP. Thus, it is shown for the first time that a confirmed defect in the GTP hydrolase activity of MnmE results in the lack of its tRNA modifying function. Moreover, the mutational analysis of the GTPase domain indicates that MnmE is closer to classical GTPases than to GTP-specific metabolic enzymes. Therefore, we propose that MnmE uses a conformational change associated with GTP hydrolysis to promote the tRNA modification reaction, in which the C-terminal Cys may function as a catalytic residue. We demonstrate that point mutations abolishing the tRNA modifying function of MnmE confer synthetic lethality, which stresses the importance of this function in the mRNA decoding process. PMID- 12730229 TI - A phospholipase D-dependent process forms lipid droplets containing caveolin, adipocyte differentiation-related protein, and vimentin in a cell-free system. AB - We developed a microsome-based, cell-free system that assembles newly formed triglyceride (TG) into spherical lipid droplets. These droplets were recovered in the d Kv1.1 for both parameters (Zhu, J., Watanabe, I., Gomez, B., and Thornhill, W. B. (2001) J. Biol. Chem. 276, 39419-39427). This previous study identified determinants in the outer pore region of Kv1.4 and Kv1.1 that positively and negatively, respectively, affected these events when expressed as homomers. Here we investigated which subunit exhibited positive or negative effects on these processes when expressed as heteromers. Kv1.4/Kv1.1 heteromers, by coexpression or expression as tandem-linked heteromers, were expressed on the cell surface at approximately 20-fold lower levels versus Kv1.4 homomers but they were trans-Golgi glycosylated. The lower Kv1.4/Kv1.1 expression level was not rescued by Kvbeta 2.1 subunits. Thus Kv1.1 inhibited high cell surface expression and partially retained the heteromer in the endoplasmic reticulum, whereas Kv1.4 stimulated trans-Golgi glycosylation. The subunit determinants and cellular events responsible for these differences were investigated. In a Kv1.4/Kv1.1 heteromer, the Kv1.1 pore was a major negative determinant, and it inhibited high cell surface expression because it induced high partial endoplasmic reticulum retention and it decreased protein stability. Other Kv1.1 regions also inhibited high surface expression of heteromers. The Kv1.1 C terminus induced partial Golgi retention and contributed to a decreased protein stability, whereas the Kv1.1 N terminus contributed to only a decreased protein stability. Thus a neuron may regulate its cell surface K+ channel protein levels by different Kv1 subfamily homomeric and heteromeric combinations that affect intracellular retention characteristics and protein stability. PMID- 12730234 TI - Impaired intracellular energetic communication in muscles from creatine kinase and adenylate kinase (M-CK/AK1) double knock-out mice. AB - Previously we demonstrated that efficient coupling between cellular sites of ATP production and ATP utilization, required for optimal muscle performance, is mainly mediated by the combined activities of creatine kinase (CK)- and adenylate kinase (AK)-catalyzed phosphotransfer reactions. Herein, we show that simultaneous disruption of the genes for the cytosolic M-CK- and AK1 isoenzymes compromises intracellular energetic communication and severely reduces the cellular capability to maintain total ATP turnover under muscle functional load. M-CK/AK1 (MAK=/=) mutant skeletal muscle displayed aberrant ATP/ADP, ADP/AMP and ATP/GTP ratios, reduced intracellular phosphotransfer communication, and increased ATP supply capacity as assessed by 18O labeling of [Pi] and [ATP]. An analysis of actomyosin complexes in vitro demonstrated that one of the consequences of M-CK and AK1 deficiency is hampered phosphoryl delivery to the actomyosin ATPase, resulting in a loss of contractile performance. These results suggest that MAK=/= muscles are energetically less efficient than wild-type muscles, but an apparent compensatory redistribution of high-energy phosphoryl flux through glycolytic and guanylate phosphotransfer pathways limited the overall energetic deficit. Thus, this study suggests a coordinated network of complementary enzymatic pathways that serve in the maintenance of energetic homeostasis and physiological efficiency. PMID- 12730236 TI - Affinity purification of ribosomes with a lethal G2655C mutation in 23 S rRNA that affects the translocation. AB - A method for preparation of Escherichia coli ribosomes carrying lethal mutations in 23 S rRNA was developed. The method is based on the site-directed incorporation of a streptavidin binding tag into functionally neutral sites of the 23 S rRNA and subsequent affinity chromatography. It was tested with ribosomes mutated at the 23 S rRNA position 2655 (the elongation factor (EF)-G binding site). Ribosomes carrying the lethal G2655C mutation were purified and studied in vitro. It was found in particular that this mutation confers strong inhibition of the translocation process but only moderately affects GTPase activity and binding of EF-G. PMID- 12730235 TI - Direct interaction of Rnd1 with Plexin-B1 regulates PDZ-RhoGEF-mediated Rho activation by Plexin-B1 and induces cell contraction in COS-7 cells. AB - Plexins are receptors for the axon guidance molecule semaphorins, and several lines of evidence suggest that Rho family small GTPases are implicated in the downstream signaling of Plexins. Recent studies have demonstrated that Plexin-B1 activates RhoA and induces growth cone collapse through Rho-specific guanine nucleotide exchange factor PDZ-RhoGEF. Here we show that Rnd1, a member of Rho family GTPases, directly interacted with the cytoplasmic domain of Plexin-B1. In COS-7 cells, coexpression of Rnd1 and Plexin-B1 induced cell contraction in response to semaphorin 4D (Sema4D), a ligand for Plexin-B1, whereas expression of Plexin-B1 alone or coexpression of Rnd1 and a Rnd1 interaction-defective mutant of Plexin-B1 did not. The Sema4D-induced contraction in Plexin-B1/Rnd1-expressing COS-7 cells was suppressed by dominant negative RhoA, a Rho-associated kinase inhibitor, a dominant negative form of PDZ-RhoGEF, or deletion of the carboxyl terminal PDZ-RhoGEF-binding region of Plexin-B1, indicating that the PDZ RhoGEF/RhoA/Rho-associated kinase pathway is involved in this morphological effect. We also found that Rnd1 promoted the interaction between Plexin-B1 and PDZ-RhoGEF and thereby dramatically potentiated the Plexin-B1-mediated RhoA activation. We propose that Rnd1 plays an important role in the regulation of Plexin-B1 signaling, leading to Rho activation during axon guidance and cell migration. PMID- 12730237 TI - Protein 14-3-3sigma interacts with and favors cytoplasmic subcellular localization of the glucocorticoid receptor, acting as a negative regulator of the glucocorticoid signaling pathway. AB - The glucocorticoid receptor (GR) alpha interacts with the highly conserved 14-3-3 family proteins. The latter bind phosphorylated serine/threonine residues of "partner" molecules and influence many signal transduction events by altering their subcellular localization and/or protecting them from proteolysis. To examine the physiologic role of 14-3-3 on the glucocorticoid-signaling pathway, we studied the nucleocytoplasmic shuttling and transactivation properties of GRalpha in a cell line replete with or devoid of 14-3-3sigma. We found that endogenous 14-3-3sigma helped localize green fluorescent protein-fused GRalpha in the cytoplasm in the absence of ligand and potentiated its nuclear export after ligand withdrawal. 14-3-3sigma also suppressed the transcriptional activity of GRalpha on a glucocorticoid-responsive promoter. Disruption of the classic nuclear export signal of 14-3-3sigma inactivated its ability to influence the nucleocytoplasmic trafficking and transactivation activity of GRalpha, whereas introduction of a mutation inactivating the binding activity of 14-3-3sigma to some of its partner proteins did not. 14-3-3sigma bound the ligand-binding domain of GRalpha through its COOH-terminal portion, in a partially ligand-dependent fashion, while it did not interact with "ligand-binding domain" of GRbeta at all. These results suggest that 14-3-3sigma functions as a negative regulator in the glucocorticoid signaling pathway, possibly by shifting the subcellular localization/circulation of this receptor toward the cytoplasm through its nuclear export signal. Since 14-3-3 proteins play significant roles in numerous cellular activities, such as cell cycle progression, growth, differentiation, and apoptosis, these actions might indirectly influence the transcriptional activity of GRalpha. Conversely, through its 14-3-3 protein interactions, GRalpha may influence these processes. PMID- 12730238 TI - Functionally important calmodulin-binding sites in both NH2- and COOH-terminal regions of the cone photoreceptor cyclic nucleotide-gated channel CNGB3 subunit. AB - Whereas an important aspect of sensory adaptation in rod photoreceptors and olfactory receptor neurons is thought to be the regulation of cyclic nucleotide gated (CNG) channel activity by calcium-calmodulin (Ca2+-CaM), it is not clear that cone photoreceptor CNG channels are similarly modulated. Cone CNG channels are composed of at least two different subunit types, CNGA3 and CNGB3. We have investigated whether calmodulin modulates the activity of these channels by direct binding to the CNGB3 subunit. Heteromeric channels were formed by co expression of human CNGB3 with human CNGA3 subunits in Xenopus oocytes; CNGB3 subunits conferred sensitivity to regulation by Ca2+-CaM, whereas CaM regulation of homomeric CNGA3 channels was not detected. To explore the mechanism underlying this regulation, we localized potential CaM-binding sites in both NH2- and COOH terminal cytoplasmic domains of CNGB3 using gel-overlay and glutathione S transferase pull-down assays. For both sites, binding of CaM depended on the presence of Ca2+. Individual deletions of either CaM-binding site in CNGB3 generated channels that remained sensitive to regulation by Ca2+-CaM, but deletion of both together resulted in heteromeric channels that were not modulated. Thus, both NH2- and COOH-terminal CaM-binding sites in CNGB3 are functionally important for regulation of recombinant cone CNG channels. These studies suggest a potential role for direct binding and unbinding of Ca2+-CaM to human CNGB3 during cone photoreceptor adaptation and recovery. PMID- 12730239 TI - Increased AMP:ATP ratio and AMP-activated protein kinase activity during cellular senescence linked to reduced HuR function. AB - Cytoplasmic export of the RNA-binding protein HuR, a process that critically regulates its function, was recently shown to be inhibited by the AMP-activated protein kinase (AMPK). In the present investigation, treatment of human fibroblasts with AMPK activators such as 5-amino-imidazole-4-carboxamide riboside, antimycin A, and sodium azide inhibited cell growth and lowered the expression of proliferative genes. As anticipated, AMPK activation also decreased both the cytoplasmic HuR levels and the association of HuR with target radiolabeled transcripts encoding such proliferative genes. HuR function was previously shown to be implicated in the maintenance of a "young cell" phenotype in models of replicative cellular senescence. We therefore postulated that AMPK activation in human fibroblasts might contribute to the implementation of the senescence phenotype through mechanisms that included a reduction in HuR cytoplasmic presence. Indeed, AMP:ATP ratios were 2-3-fold higher in senescent fibroblasts compared with young fibroblasts. Accordingly, in vitro senescence was accompanied by a marked elevation in AMPK activity. Evidence that increased AMPK activity directly contributed to the implementation of the senescent phenotype was obtained through two experimental approaches. First, use of AMPK activators triggered senescence characteristics in fibroblasts, such as the acquisition of senescence-associated beta-galactosidase (beta-gal) activity and increased p16INK4a expression. Second, infection of cells with an adenoviral vector that expresses active AMPK increased senescence-associated beta-gal activity, whereas infection with an adenovirus that expresses dominant-negative AMPK decreased senescence-associated beta-gal activity. Together, our results indicate that AMPK activation can cause premature fibroblast senescence through mechanisms that likely involve reduced HuR function. PMID- 12730240 TI - Physical and genetic interactions of cytosolic malate dehydrogenase with other gluconeogenic enzymes. AB - A truncated form (deltanMDH2) of yeast cytosolic malate dehydrogenase (MDH2) lacking 12 residues on the amino terminus was found to be inadequate for gluconeogenic function in vivo because the mutant enzyme fails to restore growth of a Deltamdh2 strain on minimal medium with ethanol or acetate as the carbon source. The DeltanMDH2 enzyme was also previously found to be refractory to the rapid glucose-induced inactivation and degradation observed for authentic MDH2. In contrast, kinetic properties measured for purified forms of MDH2 and deltanMDH2 enzymes are very similar. Yeast two-hybrid assays indicate weak interactions between MDH2 and yeast phosphoenolpyruvate carboxykinase (PCK1) and between MDH2 and fructose-1,6-bisphosphatase (FBP1). These interactions are not observed for deltanMDH2, suggesting that differences in cellular function between authentic and truncated forms of MDH2 may be related to their ability to interact with other gluconeogenic enzymes. Additional evidence was obtained for interaction of MDH2 with PCK1 using Hummel-Dreyer gel filtration chromatography, and for interactions of MDH2 with PCK1 and with FBP1 using surface plasmon resonance. Experiments with the latter technique demonstrated a much lower affinity for interaction of deltanMDH2 with PCK1 and no interaction between deltanMDH2 and FBP1. These results suggest that the interactions of MDH2 with other gluconeogenic enzymes are dependent on the amino terminus of the enzyme, and that these interactions are important for gluconeogenic function in vivo. PMID- 12730241 TI - Two new substrates in insulin signaling, IRS5/DOK4 and IRS6/DOK5. AB - We have identified two new human genes that encode proteins with tandem pleckstrin homology-phosphotyrosine binding (PH-PTB) domains at their amino termini. Because the other known PH-PTB proteins (insulin receptor substrates: IRS-1, IRS-2, IRS-3, and IRS-4, and the downstream of kinases: DOK-1, DOK-2, and DOK-3) are substrates of insulin and insulin-like growth factor (IGF)-1 receptors, we asked whether these new proteins, termed IRS5/DOK4 and IRS6/DOK5, might also have roles in insulin and IGF-1 signaling. Northern analyses indicate that IRS5/DOK4 is ubiquitously expressed but most abundant in kidney and liver. IRS6/DOK5 expression is highest in skeletal muscle. Both proteins are tyrosine phosphorylated in response to insulin and IGF-1 in transfected cells, although the kinetics differ. Insulin receptor-phosphorylated IRS5/DOK4 associates with RasGAP, Crk, Src, and Fyn, but not phosphatidylinositol 3-kinase p85, Grb2, SHP 2, Nck, or phospholipase Cgamma Src homology 2 domains, and activates MAPK in cells. IRS6/DOK5 neither associates with these Src homology 2 domains nor activates MAPK. IRS5/DOK4 and IRS6/DOK5 represent two new signaling proteins with potential roles in insulin and IGF-1 action. PMID- 12730242 TI - Hyperosmotic stress inhibits insulin receptor substrate-1 function by distinct mechanisms in 3T3-L1 adipocytes. AB - In 3T3-L1 adipocytes, hyperosmotic stress was found to inhibit insulin signaling, leading to an insulin-resistant state. We show here that, despite normal activation of insulin receptor, hyperosmotic stress inhibits both tyrosine phosphorylation of insulin receptor substrate-1 (IRS-1) and IRS-1-associated phosphoinositide 3 (PI 3)-kinase activity in response to physiological insulin concentrations. Insulin-induced membrane ruffling, which is dependent on PI 3 kinase activation, was also markedly reduced. These inhibitory effects were associated with an increase in IRS-1 Ser307 phosphorylation. Furthermore, the mammalian target of rapamycin (mTOR) inhibitor rapamycin prevented the osmotic shock-induced phosphorylation of IRS-1 on Ser307. The inhibition of mTOR completely reversed the inhibitory effect of hyperosmotic stress on insulin induced IRS-1 tyrosine phosphorylation and PI 3-kinase activation. In addition, prolonged osmotic stress enhanced the degradation of IRS proteins through a rapamycin-insensitive pathway and a proteasome-independent process. These data support evidence of new mechanisms involved in osmotic stress-induced cellular insulin resistance. Short-term osmotic stress induces the phosphorylation of IRS 1 on Ser307 by an mTOR-dependent pathway. This, in turn, leads to a decrease in early proximal signaling events induced by physiological insulin concentrations. On the other hand, prolonged osmotic stress alters IRS-1 function by inducing its degradation, which could contribute to the down-regulation of insulin action. PMID- 12730243 TI - Senescent phenotype can be reversed by reduction of caveolin status. AB - Hyporesponsiveness to growth factors is one of the fundamental characteristics of senescent cells. We previously reported that the up-regulation of caveolin attenuates the growth factor response and the subsequent downstream signal cascades in senescent human diploid fibroblasts. Therefore, in the present experiment, we investigated the modulation of caveolin status in senescent cells to determine the effect of caveolin on mitogenic signaling efficiency and cell cycling. We reduced the level of caveolin-1 in senescent human diploid fibroblasts using its antisense oligonucleotides and small interfering RNA, and this resulted in the restoration of normal growth factor responses such as the increased phosphorylation of Erk, the nuclear translocation of p-Erk, and the subsequent activation of p-Elk upon epidermal growth factor stimulation. Moreover, DNA synthesis and the re-entry of senescent cells into cell cycle were resumed upon epidermal growth factor stimulation concomitantly with decreases in p53 and p21. Taken together, we conclude that the loss of mitogenic signaling in senescent cells is strongly related to their elevated levels of caveolin-1 and that the functional recovery of senescent cells at least in the terms of growth factor responsiveness and cell cycle entry might be achieved simply by lowering the caveolin level. PMID- 12730244 TI - Biochemical characterization of yeast mitochondrial Grx5 monothiol glutaredoxin. AB - Grx5 is a yeast mitochondrial protein involved in iron-sulfur biogenesis that belongs to a recently described family of monothiolic glutaredoxin-like proteins. No member of this family has been biochemically characterized previously. Grx5 contains a conserved cysteine residue (Cys-60) and a non-conserved one (Cys-117). In this work, we have purified wild type and mutant C60S and C117S proteins and characterized their biochemical properties. A redox potential of -175 mV was calculated for wild type Grx5. The pKa values obtained by titration of mutant proteins with iodoacetamide at different pHs were 5.0 for Cys-60 and 8.2 for Cys 117. When Grx5 was incubated with glutathione disulfide, a transient mixed disulfide was formed between glutathione and the cystein 60 of the protein because of its low pKa. Binding of glutathione to Cys-60 promoted a decrease in the Cys-117 pKa value that triggered the formation of a disulfide bond between both cysteine residues of the protein, indicating that Cys-117 plays an essential role in the catalytic mechanism of Grx5. The disulfide bond in Grx5 could be reduced by GSH but at a rate at least 20 times slower than that observed for the reduction of glutaredoxin 1 from E. coli, a dithiolic glutaredoxin. This slow reduction rate could suggest that GSH may not be the physiologic reducing agent of Grx5. The fact that wild type Grx5 efficiently reduced a glutathiolated protein used as a substrate indicated that Grx5 may act as a thiol reductase inside the mitochondria. PMID- 12730245 TI - Age-related changes in collagen, pyridinoline, and deoxypyridinoline in normal human thoracic intervertebral discs. AB - Human thoracic discs were analyzed for collagen and collagen cross-links to determine the distribution due to segmental, age, and gender influences. Thoracic discs from 26 cadaveric spines (1 to 90 years old) were graded macroscopically, then separated into anular and nuclear samples. Only grade I (i.e., normal) disc samples were selected (n=209). Pyridinoline and deoxypyridinoline cross-links were initially separated by column chromatography and analyzed by reverse-phase high-pressure liquid chromatography. The collagen content was lower and the extent of pyridinoline and deoxypyridinoline were significantly higher in the nucleus compared with the anulus (p <.001). The collagen content and extent of pyridinoline were significantly lower with increasing age in the anulus and nucleus (p <.001). Young male discs had a significantly higher extent of pyridinoline compared with females (p <.001). Age, gender, and disc region differences were found to have a significant influence on the biochemical composition of the normal disc extracellular matrix. PMID- 12730246 TI - Dietary restriction does not protect the nigrostriatal dopaminergic pathway of older animals from low-dose MPTP-induced neurotoxicity. AB - To determine whether reduced caloric intake affects the susceptibility of nigrostriatal dopamine (DA) neurons to 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP)-induced neurotoxicity, 1-year-old male C57BL6 mice were offered food ad libitum or were given only 60% of the normal dietary intake. After 3 months, both groups were treated with low cumulative doses of 0, 10, 15, or 20 mg/kg MPTP. One week later, the striata were collected and DA, dihydroxyphenylalanine (DOPAC), and norepinephrine (NE) were measured. Treatment with MPTP had no effect on striatal NE but produced a dose-related depletion of DA and DOPAC in both the ad libitum-fed and the dietary-restricted mice. The MPTP induced depletions of DA and DOPAC were not ameliorated in the dietary-restricted versus the ad libitum-fed mice. Baseline DA levels and those observed after treatment with the 15-mg/kg dose of MPTP were lower in the dietary-restricted mice compared with the ad libitum-fed mice. Overall, these results suggest that, at least in 1-year-old mice, dietary restriction for 3 months does not protect nigral DA nerve terminals from low toxic dosages of MPTP. PMID- 12730247 TI - Effect of age on susceptibility to azoxymethane-induced colonic aberrant crypt foci formation in C57BL/6JNIA mice. AB - To determine the effect of age on susceptibility to azoxymethane (AOM)-induced colonic aberrant crypt foci (ACF) formation and its underlying mechanism, young and old mice were injected with AOM weekly for 4 or 5 weeks and euthanized 5 or 6 weeks later. Given the same (12 or 15) mg/kg body weight dose of AOM, old mice had significantly more ACF than young mice. However, given the same total dose of AOM (to avoid confounding effect of higher dose to heavier old mice), at a low total dose (1.5 mg) there was no age difference, but at higher total doses (1.8 and 2.2 mg) young mice had significantly more ACF than old mice. These results indicate that the age-related susceptibility to AOM differs depending on whether administration of the carcinogen is based on weight or total dose. These age differences are not due to variations in cyclooxygenase-2 expression, cell proliferation, or AOM hydroxylase activity. PMID- 12730248 TI - Androgens and women at the menopause and beyond. PMID- 12730249 TI - The positives of negatives: clinical implications of eccentric resistance exercise in old adults. PMID- 12730250 TI - The positive effects of negative work: increased muscle strength and decreased fall risk in a frail elderly population. AB - BACKGROUND: The objective of this study was to determine if a chronic eccentric training intervention, i.e., negative work, could limit or even reverse sarcopenia and its related impairments and functional limitations. Is high-force eccentric training tolerable by elderly people and will it result in improved muscle size, strength, balance, and fall risk? METHODS: 21 frail elderly subjects (mean age, 80 years) experienced 11 weeks of lower extremity resistance training. The experimental eccentric (ECC) group (n=11) performed negative work while exercising on a high-force eccentric ergometer. The active "controls" performed traditional (TRAD) (n=10) lower extremity resistance exercises (weight training). Muscle fiber cross-sectional area and strength, balance, stair descending abilities, and fall risk were assessed prior to and following this intervention. RESULTS: All ECC subjects who started the negative work intervention completed the study and reported the training to be relatively effortless; they experienced minimal and transient muscle soreness. Both groups experienced a significant increase in muscle fiber cross-sectional area (ECC=60%, TRAD=41%). Only the ECC group experienced significant improvements in strength (60%), balance (7%), and stair descent (21%) abilities. The timed up and go task improved in both groups, but only the ECC group went from a high to a low fall risk. CONCLUSIONS: These data demonstrate that lower extremity resistance exercise can improve muscle structure and function in those with limited exercise tolerance. The greater strength increase following negative work training resulted in improved balance, stair descent, and fall risk only in the ECC group. Because low energy cost is coupled to high force production with eccentric exercise, this intervention may be useful for a number of patients that are otherwise unable to achieve high muscle forces with traditional resistance exercise. PMID- 12730251 TI - Cardiovascular advantages among the offspring of centenarians. AB - BACKGROUND: A significant component of the ability to survive to exceptional old age may be familial. This study assessed the prevalence of age-related diseases in the offspring of centenarians. METHODS: The health histories of centenarian offspring (n=177) and controls (n=166) were assessed from 1997-2000 using a cross sectional study design. The offspring of 192 centenarian subjects enrolled in the nationwide New England Centenarian Study were recruited and enrolled. Controls consisted of offspring whose parents were born in the same years as the centenarians but at least 1 of whom died at an average life expectancy. Prevalence of age-related diseases including heart disease, hypertension, diabetes, cancer, osteoporosis, stroke, dementia, cataracts, glaucoma, macular degeneration, depression, Parkinson's disease, thyroid disease, and chronic obstructive pulmonary disease were compared between the two groups. RESULTS: Centenarian offspring had a 56% reduced relative prevalence of heart disease (odds ratio [OR] 0.44, 95% confidence interval [CI] 0.24, 0.80), a 66% reduced relative prevalence of hypertension (OR 0.34, 95% CI 0.21, 0.55), and 59% reduced relative prevalence of diabetes (OR 0.41, 95% CI 0.15, 1.12) after multivariate adjusted analyses. CONCLUSIONS: The offspring of centenarians demonstrate a markedly reduced prevalence of diseases associated with aging, in particular for cardiovascular disease and cardiovascular risk factors. Along with their parents, the centenarian offspring, who are in their 70s and 80s, may prove to be a valuable cohort to study genetic and environmental factors conducive to the ability to live to very old age in good health. PMID- 12730252 TI - Utilization of lipid-lowering drugs in elderly persons with increased serum low density lipoprotein cholesterol associated with coronary artery disease, symptomatic peripheral arterial disease, prior stroke, or diabetes mellitus before and after an educational program on dyslipidemia treatment. AB - BACKGROUND: We report the prevalence of use of lipid-lowering drugs in elderly persons with increased serum low-density lipoprotein (LDL) cholesterol associated with coronary artery disease (CAD), symptomatic peripheral arterial disease (PAD), prior stroke, and diabetes mellitus before and after an educational program on the treatment of dyslipidemia. METHODS: In an academic nursing home, we investigated the prevalence of use of lipid-lowering drugs in persons, mean age 77 +/- 9 years (40% men and 60% women), with a serum LDL cholesterol > or = 100 mg/dl associated with the aforementioned ailments before and after a 5-month educational program on the treatment of dyslipidemia was given to physicians and nurse practitioners. RESULTS: After the educational program, the prevalence of use of lipid-lowering drugs to treat the targeted elderly population increased (p <.001) in persons with CAD from 29% (18 of 63 persons) to 70% (44 of 63 persons), in persons with symptomatic PAD from 28% (5 of 18 persons) to 79% (15 of 19 persons), in persons with prior stroke from 24% (11 of 45 persons) to 64% (28 of 44 persons), and in diabetics from 26% (14 of 53 persons) to 67% (35 of 52 persons). CONCLUSION: A 5-month educational program on dyslipidemia treatment given to physicians and nurse practitioners in an academic nursing home improved the prevalence of use of lipid-lowering drugs in persons with increased serum LDL cholesterol associated with CAD, symptomatic PAD, prior stroke, and diabetes mellitus. PMID- 12730254 TI - Delirium among newly admitted postacute facility patients: prevalence, symptoms, and severity. AB - BACKGROUND: Delirium may persist for weeks or months, and discharging elderly patients quickly from acute care facilities is not uncommon. Therefore, the adverse impact of delirium on loss of independence may occur in the postacute setting rather than in the hospital. The purpose of this study is to describe the prevalence of delirium, delirium symptoms, and severity assessed at admission to postacute facilities. METHODS: Subjects were recruited from seven Boston-area skilled nursing facilities specializing in postacute care. Assessment instruments included the Mini-Mental Status Exam, Delirium Symptom Interview, Memorial Delirium Assessment Scale, and Confusion Assessment Method (CAM) Diagnostic Algorithm. Delirium status was categorized into four groups: full, two or more symptoms, one symptom, and no delirium. Descriptive statistics were calculated and chi-square analyses and an analysis-of-variance were used to examine delirium characteristics by delirium group. RESULTS: Among 2158 subjects, approximately 16% had full CAM-defined delirium at admission to the postacute facility. In addition, nearly 13% of the subjects had two or more symptoms of delirium, approximately 40% had one delirium symptom, and 32% had no symptoms of delirium. In a comparison of the group with no symptoms of delirium with that with CAM defined delirium, there was a significant trend toward (a) older age, (b) lower scores on the Mini-Mental Status Exam, (c) more Delirium Symptom Interview symptoms, and (d) higher Memorial Delirium Assessment Scale Scores. CONCLUSIONS: Results indicate that 16% of admissions to postacute facilities have CAM-defined delirium, and over two thirds had at least one delirium symptom. It is not known whether or not postacute staff have the training necessary to detect or manage delirium. Managing delirium may require different strategies and techniques in a postacute setting, thereby requiring further research. PMID- 12730253 TI - Prevalence of sarcopenia and predictors of skeletal muscle mass in nonobese women who are long-term users of estrogen-replacement therapy. AB - BACKGROUND: Sarcopenia refers to the loss of skeletal muscle mass with age. We have found a prevalence of sarcopenia of 22.6% in older postmenopausal women not receiving estrogen. The objective of this study was to determine the prevalence of sarcopenia in a population of older, nonobese, community-dwelling women who had been long-term users of estrogen replacement therapy (ERT). METHODS: We measured appendicular skeletal muscle mass by dual x-ray absorptiometry (DXA) in 189 women aged 59 to 78 years old who had been using ERT for at least 2 years (mean +/- SD duration, 12.7 +/- 8.2 years). We defined sarcopenia as an adjusted appendicular skeletal muscle mass (ASM) (mass divided by height squared) more than 2 SDs below the mean for a young healthy reference population. Health and menopause history were obtained. Body mass index (BMI) was calculated, and physical activity and performance were measured using the Physical Activity Scale in the Elderly, the chair rise time, the 6-minute walk, and measures of lower extremity strength and power. Serum estrone, estradiol, testosterone, and sex hormone binding globulin were measured. RESULTS: The prevalence of sarcopenia in nonobese, community-dwelling women who were long-term ERT users was 23.8%. Skeletal muscle mass correlated significantly with BMI, age at the time of starting ERT, hand grip strength, lower extremity strength and power, and testosterone level, but not with estradiol level. In linear regression analysis, BMI, leg press strength, and testosterone level contributed to adjusted ASM, accounting for 48.7% of the variance (p <.001). CONCLUSIONS: Sarcopenia is as common in nonobese women who are long-term ERT users as in community-dwelling women not using ERT, suggesting that ERT does not protect against the muscle loss of aging. BMI, strength, and testosterone level contributed to appendicular skeletal mass in women. These data suggest that interventions to target nutrition, strength training, and testosterone replacement should be further investigated for their role in preventing muscle loss with age. PMID- 12730255 TI - Acceleration patterns of the head and pelvis when walking are associated with risk of falling in community-dwelling older people. AB - BACKGROUND: A large proportion of falls in older people occur when walking, however the mechanisms underlying impaired balance during gait are poorly understood. This study evaluated acceleration patterns of the head and pelvis when walking on a level and an unpredictably irregular surface to determine whether older people at risk of falling demonstrate an impaired ability to stabilize the body under challenging conditions. METHODS: One hundred community dwelling older people aged between 75 and 93 years were evaluated for their risk of falling using a range of physiological tests previously found to be accurate predictors of falling in prospective studies. Temporo-spatial gait parameters and acceleration patterns at the head and pelvis were then measured in three orthogonal planes while subjects walked on a flat corridor and an unpredictably irregular walkway. Harmonic ratios of head and pelvis accelerations in each plane were calculated to provide an indicator of stability. RESULTS: Subjects with a high risk of falling exhibited reduced temporo-spatial gait parameters and increased step timing variability. Harmonic ratios of acceleration patterns were reduced at the head and pelvis in the vertical and antero-posterior directions. These differences were particularly evident when walking on the irregular surface. CONCLUSION: Older people at risk of falling adopt a more conservative basic walking pattern, but this does not ensure that the movements of the head and pelvis are stable. The irregular pelvis and head accelerations evident in the high risk group suggests that these subjects may have difficulty controlling trunk motion and maintaining a stable visual field when walking, particularly on irregular terrain. PMID- 12730256 TI - Old adults perform activities of daily living near their maximal capabilities. AB - BACKGROUND: Old adults' ability to execute activities of daily living (ADLs) declines with age. One possible reason for this decline is that the execution of customary motor tasks requires a substantially greater effort in old compared with young adults relative to their available maximal capacity. METHODS: We tested the hypothesis that the relative effort (i.e., the percentage of joint moment relative to maximal joint moment) to execute ADLs is higher in old adults compared with young adults. Healthy young adults (n = 13; mean age, 22 years) and old adults (n = 14; mean age, 74 years) ascended and descended stairs and rose from a chair and performed maximal-effort isometric supine leg press. Using inverse dynamics analysis, we determined knee joint moments in ADLs and computed relative effort. RESULTS: Compared with young adults, old adults had 60% lower maximal leg press moments, 53% slower knee angular velocity at peak torque, and 27% lower knee joint moments in the ADLs (all p <.05). Relative effort in ascent was 54% (SD +/- 16%) and 78% (+/-20%) in young and old adults, respectively; in descent, it was 42% (+/-20%) and 88% (+/-43%); and in chair rise, it was 42% (+/ 19%) and 80% (+/-34%) (all p <.05). The relative electromyographic activity of the vastus lateralis and the coactivity of the biceps femoris associated with this relative effort were, respectively, 2- and 1.6-fold greater in old compared with young adults in the 3 ADLs (p <.05). CONCLUSIONS: For healthy old adults, the difficulty that arises while performing ADLs may be due more to working at a higher level of effort relative to their maximum capability than to the absolute functional demands imposed by the task. PMID- 12730259 TI - Older patients' perceptions of quality of chronic knee or hip pain: differences by ethnicity and relationship to clinical variables. AB - BACKGROUND: There is marked ethnic or racial disparity in the utilization of joint replacement for osteoarthritis. The reasons are not known. Pain is the reason most patients with osteoarthritis seek care. Cultural and psychosocial factors influence how patients experience and express pain. We examined whether patient descriptions of chronic pain vary by ethnicity and if they correlate with important clinical measures used in arthritis care. METHODS: Sample consisted of 300 male veterans who were > or =50 years of age with moderate to severe symptomatic knee or hip osteoarthritis. Structured surveys were used to assess patient descriptions of pain and to collect important demographic, clinical, and psychosocial variables. Factor analysis was used to assess patterns of pain description in a comparison of African-American and Caucasian patients. Pearson correlations were used to examine relationships between pain descriptions and clinical variables. RESULTS: The two groups were similar with respect to age and other baseline clinical characteristics. A confirmatory factor analysis on quality of pain description showed that a four-factor model converged for Caucasian patients (chi square = 39.6, comparative fit index = 0.95, Tucker Lewis index = 0.93, and root mean square error of approximation = 0.047), but a three factor model was supported by the data for African-American patients (chi square= 25.4, comparative fit index = 1.00, Tucker Lewis index = 1.05, and root mean square error of approximation < or = 0.001). Chronic pain quality descriptions correlate significantly with Western Ontario and McMaster Universities Arthritis Index scores but not with radiologic stage of disease. CONCLUSIONS: African American and Caucasian elderly patients with chronic knee or hip symptomatic osteoarthritis describe the quality of their pain differently. Patient descriptions of quality of chronic knee or hip pain do not correlate with radiologic stage of disease. PMID- 12730258 TI - Elective primary total hip arthroplasty in octogenarians. AB - BACKGROUND: Total hip arthroplasty (THA) has resulted in decreased pain and increased function in people with end-stage hip arthritis for several decades. In elderly people, THA has been used most often following femoral neck fracture. However, with life spans increasing and people remaining healthy and active well into their eighties, arthroplasty surgeons are now performing primary THA as an elective procedure in octogenarians. The aim of this study was to demonstrate the efficacy of elective total hip arthroplasty in patients aged 80 years or older with end-stage hip arthritis. METHODS: Forty patients (46 THAs), aged 80 or older at the time of surgery, were identified for this study. Clinical assessment included amount of hip pain, limp, and use of assistive devices for ambulation at most recent follow-up. Radiographic assessment included implant stability, heterotopic bone formation, and osteolysis. Medical and hip-related complications, reoperations, and revisions were recorded from medical records. RESULTS: Eleven patients (27.5%) suffered a medical complication and six patients (15%) had a hip-related complication, all of which were treated and were not life threatening. Clinically, 80% were pain free and 70% walked without assistance at an average 4-year follow-up. Radiographically, all implants were stable without osteolysis. No components had been revised; however, four patients had undergone a reoperation, three for recurrent dislocation. CONCLUSIONS: Elective total hip arthroplasty is a safe and effective treatment for end-stage osteoarthritis of the hip in the elderly patient. However, the procedure is not without risk. Complications, often related to preexisting comorbidities, do occur, but mortality rates are low. PMID- 12730257 TI - Additive effects of cognitive function and depressive symptoms on mortality in elderly community-living adults. AB - BACKGROUND: Poor cognitive function and depressive symptoms are common in the elderly, frequently coexist, and are interrelated. Both risk factors are independently associated with mortality. Few studies have comprehensively described how the combination of poor cognitive function and depressive symptoms affect the risk for mortality. Our aim was to examine whether the combination of varying levels of cognitive function and depressive symptoms affect the risk of mortality in community-living elderly adults. METHODS: We studied 6301 elderly adults (mean age, 77 years; 62% women; 81% white) enrolled in the Asset and Health Dynamics Among the Oldest Old (AHEAD) study, a prospective study of community-living participants conducted from 1993 to 1995. Cognitive function and depressive symptoms were measured using two validated measures developed for the AHEAD study. On each measure, participants were divided into tertiles representing the best, middle, and worst scores, and then placed into one of nine mutually exclusive groups ranging from best functioning on both measures to worst functioning on both measures. Mortality rates were assessed in each of the nine groups. Cox proportional hazards models were used to control for potentially confounding characteristics such as demographics, education, income, smoking, alcohol consumption, comorbidity, and baseline functional impairment. RESULTS: During 2 years of follow-up, 9% (548) of the participants died. Together, cognitive function and depressive symptoms differentiated between elderly adults at markedly different risk for mortality, ranging from 3% in those with the best function on both measures to 16% in those with the worst function on both measures (p <.001). Furthermore, for each level of cognitive function, more depressive symptoms were associated with higher mortality rates, and for each level of depressive symptoms, worse cognitive function was associated with higher mortality rates. In participants with the best cognitive function, mortality rates were 3%, 5%, and 9% in participants with low, middle, and high depressive symptoms, respectively (p <.001 for trend). The corresponding rates were 6%, 7%, and 12% in participants with the middle level of cognitive function (p <.001 for trend), and 10%, 13%, and 16% in participants with the worst level of cognitive function (p <.001 for trend). After adjustment for confounders, participants with the worst function on both measures remained at considerably higher risk for death than participants with the best function on both measures (adjusted hazard ratio, 3.1; 95% confidence interval, 2.0-4.7). CONCLUSIONS: Cognitive function and depressive symptoms can be used together to stratify elderly adults into groups that have significantly different rates of death. These two risk factors are associated with an increased risk in mortality in a progressive, additive manner. PMID- 12730260 TI - "Polyherbacy": herbal supplements as a form of polypharmacy in older adults. PMID- 12730261 TI - In situ localization associates biologically active plant natriuretic peptide immuno-analogues with conductive tissue and stomata. AB - Plant natriuretic peptide immuno-analogues (irPNP) have previously been shown to affect a number of biological processes including stomatal guard cell movements, ion fluxes and osmoticum-dependent water transport. Tissue printing and immunofluorescent labelling techniques have been used here to study the tissue and cellular localization of irPNP in ivy (Hedera helix L.) and potato (Solanum tuberosum L.). Polyclonal antibodies active against human atrial natriuretic peptide (anti-hANP) and antibodies against irPNP from potato (anti-StPNP) were used for immunolabelling. Tissue prints revealed that immunoreactants are concentrated in vascular tissues of leaves, petioles and stems. Phloem-associated cells, xylem cells and parenchymatic xylem cells showed the strongest immunoreaction. Immunofluorescent microscopy with fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG supported this finding and, furthermore, revealed strong labelling to stomatal guard cells and the adjacent apoplastic space as well. Biologically active immunoreactants were also detected in xylem exudates of a soft South African perennial forest sage (Plectranthus ciliatus E. Mey ex Benth.) thus strengthening the evidence for a systemic role of the protein. In summary, in situ cellular localization is consistent with physiological responses elicited by irPNPs reported previously and is indicative of a systemic role in plant homeostasis. PMID- 12730263 TI - Genetic manipulation of glycine decarboxylation. AB - The glycine-serine interconversion, catalysed by glycine decarboxylase and serine hydroxymethyltransferase, is an important reaction of primary metabolism in all organisms including plants, by providing one-carbon units for many biosynthetic reactions. In plants, in addition, it is an integral part of the photorespiratory metabolic pathway and produces large amounts of photorespiratory CO(2) within mitochondria. Although controversial, there is significant evidence that this process, by the relocation of glycine decarboxylase within the leaves from the mesophyll to the bundle-sheath, contributed to the evolution of C(4) photosynthesis. In this review, some aspects of current knowledge about glycine decarboxylase and serine hydroxymethyltransferase and the role of these enzymes in metabolism, about the corresponding genes and their expression as well as about mutants and anti-sense plants related to these genes or processes will be summarized and discussed. From a comparison of the available information about the number and organization of GDC and SHMT genes in the genomes of Arabidopsis thaliana and Oryza sativa it appears that these and, possibly, other genes related to photorespiration, are similarly organized even in only very distantly related angiosperms. PMID- 12730262 TI - Sugars regulate cold-induced gene expression and freezing-tolerance in barley cell cultures. AB - The hypothesis that the extracellular concentration of sugars helps regulate the acclimation of plant cells to cold was tested in this work. Suspension cultures were used to control the concentration of sugars in the medium supplied to barley cell cultures (Hordeum vulgare L. cv. Igri), replacing the medium daily to help maintain the concentration. Freezing tolerance and the levels of mRNA expression of the stress-response genes blt4.9 (coding for a non- specific lipid transfer protein) and dhn1 (coding for a dehydrin) were measured. Similar levels of freezing-tolerance and gene expression were obtained in the experiments as occur during cold-acclimation in the crown of the whole plant. In the cell cultures, cold (6/2 degrees C) did not induce an increase in freezing tolerance or in the expression of detectable levels of blt4.9 or dhn1 mRNAs when only 1 g l-1 sucrose was supplied. However, the cells in this low sucrose medium in the cold were not sugar-starved, indicating that this did not explain the failure of the cells to acclimate when grown in the cold environment. Ten g l-1 sucrose supplied to cells grown in the warm (25 degrees C) induced acclimation to freezing and up regulation of expression of blt4.9 and dhn1 mRNAs. Osmolality of the medium did not explain this. Thirty g l-1 sucrose induced yet higher levels of freezing tolerance and of blt4.9 and dhn1 mRNAs in cultures grown in either the cold or the warm environment. The results implicate sugars in the regulation of cold acclimation PMID- 12730264 TI - Changing desiccation tolerance of pea embryo protoplasts during germination. AB - Protoplasts were isolated from pea (Pisum sativum L. cv. Alaska) embryonic axes during and after germination to determine whether the loss of desiccation tolerance in the embryos also occurs in the protoplasts. At all times studied, protoplast survival decreased as water content decreased; however, the sensitivity to dehydration was less when the protoplasts were isolated from embryos that were still desiccation-tolerant (12 h and 18 h of imbibition) than when protoplasts were derived from axes that were sensitive (24 h and 36 h of imbibition). The water content at which 50% of the population was killed (WC50) increased throughout germination and early seedling growth for both the intact tissue and the protoplasts derived from them. Prior to radicle emergence, protoplasts were less desiccation-tolerant than the intact axes; however, protoplasts isolated from radicles shortly after emergence had lower WC50s than the intact radicles. A comparison of protoplast survival after isolation and dehydration in either 500 mM sucrose/raffinose or 700 mM sucrose revealed no difference in tolerance except at 24 h of imbibition, when protoplasts treated in the more concentrated solution had improved tolerance of dehydration. Although intact epicotyls are generally more desiccation-tolerant than radicles, protoplasts isolated separately from epicotyls and radicles did not differ in tolerance. Collectively, these data suggest that protoplasts gradually lose desiccation tolerance during germination, as do the orthodox embryos from which they were derived. However, even prior to radicle emergence, protoplasts display a sensitivity to progressive dehydration that is similar to that shown by recalcitrant and ageing embryos. PMID- 12730265 TI - Branch junctions and the flow of water through xylem in Douglas-fir and ponderosa pine stems. AB - Water flowing through the xylem from the roots to the leaves of most plants must pass through junctions where branches have developed from the main stem. These junctions have been studied as both flow constrictions and components of a hydraulic segmentation mechanism to protect the main axes of the plant. The hydraulic nature of the branch junction also affects the degree to which branches interact and can respond to changes in flow to other branches. The junctions from shoots of two conifer species were studied, with particular emphasis on the coupling between the downstream branches. Flow was observed qualitatively by forcing stain through the junctions and the resulting patterns showed that flow into a branch was confined to just part of the subtending xylem until a considerable distance below the junction. Junctions were studied quantitatively by measuring flow rates in a branch before and after flow was stopped in an adjacent branch and by measuring the hydraulic resistance of the components of the junction. Following flow stoppage in the adjacent branch, flow into the remaining branch increased, but considerably less than predicted based on a simple resistance analogue for the branch junction that assumes the two branches are fully coupled. The branches downstream from a junction, therefore, appear to be limited in their interconnectedness and hence in their ability to interact. PMID- 12730266 TI - Vascular tissue in the stem and roots of woody plants can conduct light. AB - The role of vascular tissue in conducting light was analysed in 21 species of woody plants. Vessels, fibres (both xylem and phloem fibres) and tracheids in woody plants are shown to conduct light efficiently along the axial direction of both stems and roots, via their lumina (vessels) or cell walls (fibres and tracheids). Other components, such as sieve tubes and parenchyma cells, are not efficient axial light conductors. Investigation of the spectral properties of the conducted light indicated that far-red light was conducted most efficiently by vascular tissue. Light gradients in the axial direction were also investigated and revealed that conducted light leaked out of the light-conducting structures to the surrounding living tissues. These properties of the conducted light suggest a close relationship with metabolic activities mediated by phytochromes. The results therefore indicate not only that signals from the external light environment can enter the interior of stems above ground and are conducted by vascular tissue towards roots under ground, but also that the light conducted probably contributes directly to photomorphogenic activities within them. PMID- 12730267 TI - Growth and morphogenesis at the vegetative shoot apex of Anagallis arvensis L. AB - A non-destructive replica method and a 3-D reconstruction algorithm are used to analyse the geometry and expansion of the shoot apex surface. Surface expansion in the central zone of the apex is slow and nearly isotropic while surface expansion in the peripheral zone is more intense and more anisotropic. Within the peripheral zone, the expansion rate, expansion anisotropy, and the direction of maximal expansion vary according to the age of adjacent leaf primordia. For each plastochron, this pattern of expansion is rotated around the apex by the Fibonacci angle. Early leaf primordium development is divided into four stages: bulging, lateral expansion, separation, and bending. These stages differ in their geometry and expansion pattern. At the bulging stage, the site of primordium initiation shows an intensified expansion that is nearly isotropic. The following stages develop sharp meridional gradients of expansion rates and anisotropy. The adaxial primordium boundary inferred from the surface curvature is shifting until the separation stage, when a crease develops between the primordium and the apex dome. The cells forming the crease, i.e. the future leaf axil, expand along the axil and contract across it. Thus they are arrested in this unique position. PMID- 12730268 TI - Identification of three MADS-box genes expressed in sunflower capitulum. AB - Three cDNA clones, HaPI, HaAG and HaAP3, were isolated from sunflower inflorescences at the R2 stage of development. The cDNAs share high sequence similarity with the PISTILLATA, AGAMOUS, and APETALA3 genes from Arabidopsis, respectively, which contain a MADS-box and are involved in floral organ development. Expression of the corresponding genes was analysed by northern blots and in situ hybridization. They are expressed preferentially in the R3 and R4 stages of capitulum development. HaAG accumulates in fertile flowers, mainly in stamens, while HaPI and HaAP3 are preferentially expressed in ray (sterile) flowers and more weakly in petals and stamens of fertile flowers. PMID- 12730269 TI - Use of the pressure probe in studies of stomatal function. AB - Over the past few decades the pressure probe has been used extensively in studies of the hydromechanical and osmotic properties of plant cells. However, although pressure probe techniques have been employed successfully in the study of stomatal function, there is no detailed account of this special application of the pressure probe technique. This paper describes the construction and use of the pressure probe in studies relating to stomatal function, and reviews the current state of knowledge of stomatal function in relation to guard cell and leaf hydromechanical properties. PMID- 12730270 TI - Molecular cloning of the cDNA encoding aspartate aminotransferase from bean root nodules and determination of its role in nodule nitrogen metabolism. AB - A cDNA clone encoding aspartate aminotransferase (PVAAT-2) (EC 2.6.1.1) was isolated from the common bean Phaseolus vulgaris nodule cDNA library. The nucleotide sequence analysis of the full-length cDNA allowed its identification by comparison with sequence databases. The amino acid sequence of the bean PvAAT 2 showed high similarity with the AAT-2 isoforms described in other leguminous plants. The amino-terminal region of the PvAAT-2 contains a sequence, which shares common features of plastid transit peptides. Southern blot analysis showed that the PvAAT-2 clone is encoded by a single gene in the P. vulgaris genome. Analysis of the PvAAT-2 mRNA levels suggests that the expression of this gene is nodule enhanced. The PvAAT-2 transcript is more abundant in nodules with increased synthesis of amides and is down-regulated in conditions where ureides accumulate. When plants were supplemented with ureides or with amides, PvAAT-2 expression was reduced, while it was not affected when plants were treated with allopurinol, an inhibitor of ureide synthesis. On the other hand, the expression of asparagine synthetase (another enzyme involved in the synthesis of amides) is not affected either by ureides or amides. These data suggest a role for AAT-2 in the mechanism involved in the synthesis of nitrogen compounds in bean nodules. PMID- 12730271 TI - Vacuolar system distribution in Arabidopsis tissues, visualized using GFP fusion proteins. AB - Green fluorescent protein (GFP) allows the direct visualization of gene expression and the subcellular localization of fusion proteins in living cells. The localization of different GFP fusion proteins in the secretory system was studied in stably transformed Arabidopsis plants cv. Wassilewskaja. Secreted GFP (SGFP) and GFP retained in the ER (GFP-KDEL) confirmed patterns already known, but two vacuolar GFPs (GFP-Chi and Aleu-GFP) labelled the Arabidopsis vacuolar system for the first time, the organization of which appears to depend on cell differentiation. GFP stability in the vacuoles may depend on pH or degradation, but these vacuolar markers can, nevertheless, be used as a tool for physiological studies making these plants suitable for mutagenesis and gene-tagging experiments. PMID- 12730272 TI - Carbon dioxide action on ethylene biosynthesis of preclimacteric and climacteric pear fruit. AB - Ethylene production in pear fruit was studied at 2 degrees C. Several observations showed that the inhibiting effect of CO2 on ethylene production did not operate only via the binding site of the ethylene binding protein. Ethylene production of freshly harvested pears was stimulated by 1-methylcyclopropene (1 MCP), but unaffected or inhibited by CO2 which points to different action sites for both molecules. In climacteric pears, where ethylene production was strongly inhibited by 1-MCP, a range of applied CO2 partial pressures was able to inhibit ethylene production further, to an extent similar to untreated pears. In the case of pears that had been stored for a period of 25 weeks, CO2 only had a clear effect after 1-MCP pretreatment. Respiration measurements showed that the effect of CO2 on ethylene production did not operate via an effect on respiration. Ethylene production models based on measurements of whole pears were used to study CO2 effects. Kinetic parameters derived from the models point to the conversion from ACC to ethylene by ACC oxidase as a possible action site for CO2 inhibition. PMID- 12730273 TI - Isolation and characterization of four ethylene perception elements and their expression during ripening in pears (Pyrus communis L) with/without cold requirement. AB - Pear (Pyrus communis L.) are climacteric fruit: their ripening is associated with a burst of autocatalytic ethylene production. Some late pear cultivars, such as Passe-Crassane (PC) require a long (80 d) chilling treatment before the fruit will produce autocatalytic ethylene and ripen. As the cold requirement is linked to the capacity to respond to ethylene (or its analogue, propylene), three pear cDNAs homologous to the Arabidopsis ethylene receptor genes At-ETR1, At-ERS1, and At-ETR2, designated Pc-ETR1a (AF386509), Pc-ERS1a (AF386515), and Pc-ETR5 (AF386511), respectively, have been isolated. A pear homologue of the Arabidopsis ethylene signal transduction pathway gene At-CTR1, called Pc-CTR1 (AF386508) has also been isolated. The search of the genomic sequences for Pc-ETR1a and Pc-ERS1a resulted in the isolation of four related genomic clones Pc-DETR1a (AF386525), Pc DETR1b (AF386520), Pc-DERS1a (AF386517), and Pc-DERS1b (AF386522). Analysis of transcript levels for the four cDNAs in PC and pear fruit genotypes with little or no cold requirement revealed that Pc-ETR1a expression increased during chilling treatment, and Pc-ETR1a, Pc-ERS1a, Pc-ETR5, and Pc-CTR1 expression increased during fruit ripening and after ethylene treatment. Whether the differences in the ethylene response elements studied here are the cause or an effect of the cold requirement in PC fruit is discussed. PMID- 12730274 TI - Instrumental methods for studies of structure and function of root systems of large trees. AB - New methods using different physical principles have been successfully applied in studies of root systems of large trees. The ground-penetrating radar technique provides 3D images of coarse roots (starting with a diameter of about 20 mm) from the soil surface down to a depth of several metres. This can even be done under layers of undisturbed materials such as concrete, asphalt and water. Fine roots cannot be visualized by this method, but the total rooted volume of soil can be determined. The differential electric conductance method has been used for fast measurement of conducting (absorbing) root surfaces. However, more testing is needed. Both these methods are non-invasive. The results can be verified by an almost harmless excavation of whole root systems, including fine roots, using the ultrasonic air-stream (air-spade) method. This method is suitable for all studies, as well as practical operations on roots or objects in their vicinity, where a gentle approach is required. Sap flow measurements on their own or in tandem with soil moisture monitoring play a leading role in studying root function and hydraulic redistribution of flow in the soil. The water absorption function of roots can be studied by measuring sap flow on individual root branches directly (as on crown branches) and also indirectly, by measuring the radial pattern of sap flow in different sapwood depths at the base of a stem. Root zone architecture can also be estimated indirectly by studying its functionality. The heat field deformation method with multi-point sensors has been found to be very convenient for this purpose. A combination of several such methods is recommended whenever possible, in order to obtain detailed information about the root systems of trees. PMID- 12730275 TI - Allodynia and hyperalgesia in adjuvant-induced arthritic rats: time course of progression and efficacy of analgesics. AB - The complete Freund's adjuvant (CFA)-induced arthritic rat model has extensively served as a laboratory model in the study of arthritic pain. However, the time courses of allodynia and hyperalgesia and the efficacies of different analgesics have not fully been analyzed in this model. Mechanical allodynia, thermal and joint hyperalgesia, and other disease development parameters (body weight, mobility, paw volume, and joint stiffness) were measured on postinoculation days (PIDs) 0 to 28 in rats. Acute analgesic efficacies of drugs were evaluated on PID 9 when degrees of allodynia, hyperalgesia, and joint stiffness in the ipsilateral paw reached almost the maximum, although those in the contralateral paw changed only slightly. In the ipsilateral paw, thermal hyperalgesia reached the maximum on PID 1, whereas mechanical allodynia and joint hyperalgesia progressively developed during the first 7 or 8 days, being tuned in to arthritis development. In the contralateral paw, thermal hyperalgesia never occurred, whereas mechanical allodynia and joint hyperalgesia developed after PID 11. Morphine and tramadol had full efficacies for all the pain parameters tested at sedation-inducing doses. Indomethacin and diclofenac significantly but partially improved thermal and joint hyperalgesia. Amitriptyline significantly reduced thermal and joint hyperalgesia only at sedation-inducing dose. Acetaminophen, carbamazepine, and gabapentin had, at the most, very small efficacies. In conclusion, the present study provided integrated information about the time course of pain and other disease development parameters in the CFA-induced arthritic rats, and clarified acute efficacies of different categories of analgesics for the allodynia and hyperalgesia. PMID- 12730276 TI - Vinpocetine is a potent blocker of rat NaV1.8 tetrodotoxin-resistant sodium channels. AB - Vinpocetine is a clinically used synthetic vincamine derivative with a diverse pharmacological profile that includes action at several ion channels, principally "generic" populations of sodium channels that give rise to tetrodotoxin-sensitive conductances. A number of cell types are known to express tetrodotoxin-resistant (TTXr) sodium conductances, the molecular bases of which have remained elusive until recently. One such TTXr channel, termed NaV1.8, is of particular interest because of its prominent and selective expression in peripheral afferent nerves. The effects of vinpocetine on TTXr channels specifically, are unknown. We have assessed the effects of the drug on cloned rat NaV1.8 channels expressed in a dorsal root ganglion-derived cell line, ND7/23. Vinpocetine produced a concentration- and state-dependent inhibition of NaV1.8 sodium channel activity. Voltage-clamp experiments revealed an approximately 3-fold increase in vinpocetine potency when whole-cell NaV1.8 conductances were elicited from relatively depolarized potentials (-35 mV; IC50 = 3.5 microM) compared with hyperpolarized holding potentials (-90 mV; IC50 = 10.4 microM). Vinpocetine also produced an approximately 22 mV leftward shift in the voltage dependence of NaV1.8 channel inactivation but did not affect the voltage range of channel activation. These properties are reminiscent of several other known sodium channel blockers and suggested that vinpocetine may exhibit frequency-dependent block. Accordingly, tonic block of NaV1.8 channels by vinpocetine (3 microM) increased proportionally with increasing depolarizing commands over the frequency range 0.1 to 1Hz. In summary, the present data demonstrate that vinpocetine is capable of blocking NaV1.8 sodium channel activity and suggest a potential additional utility in various sensory abnormalities arising from abnormal peripheral nerve activity. PMID- 12730277 TI - Muscarinic cholinergic and beta-adrenergic contribution to hindquarters vasodilation and cardiac responses to cocaine. AB - Cocaine produces a pressor response associated with an initial hindquarters vasoconstriction followed by a prolonged vasodilation in conscious rats. Propranolol pretreatment prevented the vasodilation and enhanced the pressor response, whereas atropine methylbromide pretreatment reduced the increase in systemic vascular resistance. We studied the role of selective muscarinic and beta-adrenoceptor antagonists on responses to cocaine in rats with an increase in systemic vascular resistance to cocaine (vascular responders). Arterial blood pressure and ascending aortic and distal descending aortic blood flow using pulsed Doppler flowmetry were measured. In conscious rats, cocaine (5 mg/kg i.v.) elicited consistent pressor responses but variable systemic and hindquarters vascular resistance responses that were directly correlated, suggesting that skeletal muscle resistance responses comprise an important component of systemic vascular resistance. ICI 118551 [(+/-)-1-[2,3-(dihydro-7-methyl-1H-inden-4 yl)oxy]-3-[(1-methylethyl)-amino]-2-butanol] (0.5 mg/kg i.v.) pretreatment prevented the hindquarters vasodilation, enhancing the increase in systemic vascular resistance and the pressor response while further depressing the cardiac output response, similar to the effects of propranolol. Atenolol (1 mg/kg) pretreatment attenuated the stroke volume and cardiac output responses while enhancing the increase in systemic vascular resistance without affecting the hindquarters responses. In contrast, M2 antagonist methoctramine (0.3 mg/kg) pretreatment had similar effects as atropine in reducing the decrease in cardiac output by reducing the increase in systemic vascular resistance, whereas the M1 antagonist pirenzipine (0.02 mg/kg) did not alter responses. Therefore, the cocaine-induced pressor response is ameliorated by beta2-adrenoceptor mediated skeletal muscle vasodilation, whereas the decrease in cardiac output and the increase in systemic vascular resistance are dependent on M2-cholinoceptor activation. PMID- 12730278 TI - Functional characterization of human UDP-glucuronosyltransferase 1A9 variant, D256N, found in Japanese cancer patients. AB - SN-38 (7-ethyl-10-hydroxycamptothecin), an active metabolite of the antitumor prodrug irinotecan, is conjugated and detoxified to SN-38 10-O-beta-d-glucuronide by hepatic UDP-glucuronosyltransferase (UGT) 1A1. Recent studies have revealed that other UGT1A isoforms, UGT1A7 and UGT1A9, also participate in SN-38 glucuronidation. Although several genetic polymorphisms are reported for UGT1A1 and UGT1A7 that affect the SN-38 glucuronidation activities, no such polymorphisms have been identified for UGT1A9. In the present study, UGT1A9 exon 1 and its flanking regions were sequenced from 61 Japanese cancer patients who were all treated with irinotecan. A novel nonsynonymous single nucleotide polymorphism was identified in UGT1A9 exon 1, heterozygous 766G>A resulting in the amino acid substitution of D256N. The wild-type and D256N UGT1A9s were transiently expressed at similar protein levels in COS-1 cells, and their membrane fractions were characterized in vitro for the glucuronidation activities toward SN-38. The apparent Km values were 19.3 and 44.4 microM, and the Vmax values were 2.94 and 0.24 pmol/min/mg of membrane protein for the wild-type and D256N variant, respectively. The SN-38 glucuronidation efficiency (normalized Vmax/Km) of D256N was less than 5% that of wild-type UGT1A9. These results clearly indicate that the D256N variant is essentially nonfunctional with regard to SN-38 glucuronidation. These findings highlight the importance of further studies into the potential influence of UGT1A9 D256N variant to irinotecan metabolism in vivo. PMID- 12730279 TI - Pediatric sibling donors of successful and unsuccessful hematopoietic stem cell transplants (HSCT): a qualitative study of their psychosocial experience. AB - OBJECTIVE: To examine the psychosocial impact of pediatric hematopoietic stem cell transplants (HSCTs) on sibling donors, particularly in unsuccessful HSCTs. METHODS: Fifteen sibling HSCT donors, 8 who had participated in successful transplants, and 7 whose recipient sibling died, were interviewed (M age = 19.6 years at time of interviews, and 13.3 years at time of sibling's HSCT). Grounded theory methodology was used to derive themes from interviews. RESULTS: Siblings in the unsuccessful transplant group reported greater negative impact and feelings of guilt following the HSCT and were less likely to experience positive psychological effects. Both groups reported that informed consent involved "no choice" and that psychological aspects of the procedure outweighed physical aspects. CONCLUSIONS: Some themes differentiated sibling donors of successful and unsuccessful transplants, while others were common to both groups. All emergent themes reinforced the importance of providing sibling donors with developmentally appropriate, accurate information and long-term psychological support. PMID- 12730280 TI - Quality of life of adolescents with perceived disabilities. AB - OBJECTIVE: To compare the self-perceived quality of life (QoL) of adolescents with and without disabilities. METHOD: A school-based survey was conducted with 2801 7th to 12th grade students in a rural area of the United States. The Youth Quality of Life Instrument-Surveillance Module and the Youth Quality of Life Group-Disability Screener were completed. RESULTS: Twenty-one percent of all students surveyed reported having one or more physical, emotional, or learning disabilities. Adolescents with disabilities reported lower QoL than adolescent without disabilities. However, self-rated health, depressive symptoms, and contextual variables were significant covariates in the relationship between disability and QoL. CONCLUSIONS: These findings suggest channels to improve the QoL of adolescents with disabilities. Specifically, reducing social and environmental barriers to promote inclusion of adolescents with disabilities in school, family, and community activities is one practical avenue for reducing disparities in QoL. PMID- 12730281 TI - Adolescent oral sex, peer popularity, and perceptions of best friends' sexual behavior. AB - OBJECTIVE: To provided initial descriptive information regarding adolescents' engagement in oral sex and to investigate adolescents' perceptions of their best friends' sexual behavior and peer-reported popularity as two social mechanisms that may influence engagement in oral sex. METHODS: A total of 212 tenth graders reported their engagement in oral sex and intercourse, number of sexual partners, and use of sexually transmitted infection (STI) protection, as well as perceptions of their best friends' sexual behaviors. Sociometric assessment yielded peer-reported measures of adolescents' preference- and reputation-based popularity. RESULTS: Adolescents were more likely to report engagement in oral sex than intercourse, report more oral sex partners than intercourse partners, and were unlikely to report use of STI protection during oral sex. Perceptions of best friends' behavior were significantly associated with adolescents' own oral sex behavior, but not intercourse. Adolescents who reported sexual activity had high levels of reputation-based popularity, but not likeability among peers; however, sex with more partners was associated with lower levels of popularity. CONCLUSIONS: Implications for prevention programs are discussed. PMID- 12730282 TI - Long-term behavior problems following pediatric traumatic brain injury: prevalence, predictors, and correlates. AB - OBJECTIVE: To study identified rates of long-term behavior problems in children with traumatic brain injury (TBI) compared to children with only orthopedic injuries and risk factors and correlates for new behavior problems following TBI. METHODS: Sample included children with severe TBI (n = 42), moderate TBI (n = 41), and orthopedic injuries only (ORTHO;n = 50). The baseline assessment measured child behavior, adaptation, and neuropsychological, academic, and family functioning. Follow-ups were conducted at 6 and 12 months and at an extended follow-up a mean of 4 years after injury. RESULTS: The prevalence of caseness, defined as elevated behavior problem ratings, was higher in one or both TBI groups than in the ORTHO group at each follow-up (e.g., 36% of severe TBI group, 22% of moderate TBI group, and 10% of ORTHO group at extended follow-up). Most instances of postinjury-onset caseness at the extended follow-up were evident within the first year after TBI. Predictors were severe TBI, socioeconomic disadvantage, and preinjury behavioral concerns. Concurrent correlates included weakness in working memory and adaptive behavior skills, poorer behavior and school competence, and adverse family outcomes. CONCLUSIONS: Postinjury-onset caseness is persistent, risks are multifactorial, and correlates include child dysfunction and family sequelae. PMID- 12730283 TI - Discrepancies between mother and adolescent perceptions of diabetes-related decision-making autonomy and their relationship to diabetes-related conflict and adherence to treatment. AB - OBJECTIVE: To document the relationship between discrepancies in mother and adolescent perceptions of diabetes-related decision-making autonomy, diabetes related conflict, and regimen adherence. METHODS: The sample was composed of 82 mother-adolescent dyads. Measures included adolescent and mother reports of diabetes-related decision-making autonomy, diabetes-related conflict, and regimen adherence. Nurses' reports of adherence and number of glucose tests performed each day were also obtained. RESULTS: Discrepancies between mother and adolescent perceptions of decision-making autonomy were related to greater maternal report of diabetes-related conflict. In particular, when adolescents reported that they were more in charge of decisions than reported by their mothers, mothers reported more conflict. Discrepancies between mother and adolescent perceptions of decision-making autonomy were not related to regimen adherence. CONCLUSIONS: The findings suggest that discrepancies between mother and adolescent perceptions of diabetes-related decision-making autonomy may be a potentially important area for intervention. PMID- 12730284 TI - Brief report: child-rearing practices of caregivers with and without a child with juvenile rheumatoid arthritis: perspectives of caregivers and professionals. AB - OBJECTIVE: To evaluate predictions from professionals in pediatric rheumatology regarding the child-rearing practices of caregivers of children with juvenile rheumatoid arthritis (JRA) and healthy classmates. METHODS: Sixteen professionals identified items from the Child-Rearing Practices Report (CRPR) that were expected to differentiate between caregivers of children with JRA (64 mothers, 45 fathers) and caregivers of healthy classmates (64 mothers, 40 fathers). Families were interviewed, and physician ratings of disease severity were obtained. RESULTS: Experts predicted difficulties in protectiveness, discipline, and worry. Ratings from parents of children with JRA showed modest agreement with the professionals, surprising similarity to controls, and a limited association with disease factors. CONCLUSIONS: Contrary to expert opinion, JRA has only a modest influence on some child-rearing practices. Educating health care providers may minimize misperceptions about caring for children with JRA, and screening parents of children with more severe disease may assist in allocating education and services for families. PMID- 12730285 TI - Brief report: does posttraumatic stress apply to siblings of childhood cancer survivors? AB - OBJECTIVE: To investigate whether adolescent siblings of childhood cancer survivors experience posttraumatic stress (PTS). METHODS: Participants included 78 adolescent siblings of adolescent cancer survivors who completed self-report measures of anxiety, PTS, and perceptions of the cancer experience. RESULTS: Nearly half (49%) of our sample reported mild PTS and 32% indicated moderate to severe levels. One fourth of siblings thought their brother/sister would die during treatment; over half found the cancer experience scary and difficult. These perceptions were related to PTS. Siblings reported more PTS symptoms than a reference group of nonaffected teens but had similar levels of general anxiety. CONCLUSIONS: Levels of PTS are elevated for siblings of childhood cancer survivors. Thus, PTS may be a useful model for understanding siblings' long-term reactions to cancer. Future research and clinical efforts should consider the needs of siblings of childhood cancer survivors in a family context. PMID- 12730286 TI - Case study: feasibility of multisystemic therapy as a treatment for urban adolescents with poorly controlled type 1 diabetes. AB - OBJECTIVE: To determine the feasibility of using multisystemic therapy (MST), an intensive, home-based psychotherapy, to improve poor metabolic control among four adolescents with type 1 diabetes. METHOD: A multiple baseline, repeated measures design was used. Blood glucose readings were obtained every 2 weeks during a baseline period, a 7-month intervention, and a month-long postintervention period. Glycosylated hemoglobin (GHb) was measured at baseline and follow-up and was contrasted between two participants who received MST and two participants who did not receive the intervention. RESULTS: Intervention participants showed improvements in health status, as well as fewer hospitalizations and emergency room visits. CONCLUSIONS: Findings from this case study suggest that MST has the potential to improve treatment adherence and metabolic control among teens with poorly controlled type 1 diabetes. PMID- 12730288 TI - An epigenetic road map for histone lysine methylation. PMID- 12730289 TI - Trafficking of signaling modules by kinesin motors. AB - The human genome has more than 40 kinesin genes whose protein products organize intracellular traffic along microtubules. Research during the past two years has begun to elucidate the cargoes carried by kinesins and the nature of the kinesin cargo linkage. Modular protein-protein interactions connect kinesins to diverse cellular molecules, which, apart from their other functions, serve as kinesin cargo linkers. Many of these newly identified linkers are scaffolds for signaling pathways, and mounting evidence now indicates that kinesins transport pre assembled signaling modules as vesicular cargo. These findings bring together two fields, signal transduction and molecular motors, and lead to a deeper understanding of the interplay between trafficking, localization and intercellular communication. PMID- 12730290 TI - Signal pathway integration in the switch from the mitotic cell cycle to meiosis in yeast. AB - Diploid yeast, like most eukaryotes, can undergo meiotic differentiation to form haploid gametes. Meiotic differentiation and cell growth (proliferation) are mutually exclusive programs, and in yeast the switch between growth and meiosis is controlled by nutritional signals. The signaling pathways that mediate nutritional controls on meiotic initiation fall into three broad classes: those that respond to nutrient starvation, those that respond to non-fermentable carbon sources, and those that respond to glucose. At the onset of meiosis, nutritional signaling pathways converge on transcriptional regulation of two genes: IME1, which encodes a transcription factor; and IME2, which encodes a protein kinase. Transcription of IME1 and IME2 trigger initiation of meiosis, and the expression of these two genes is linked with one other, with expression of later meiotic genes and with early meiotic events such as DNA replication. In addition, the signaling pathways that control IME1 and IME2 expression are themselves integrated through a variety of mechanisms. Thus the signal network that controls the switch from growth to meiotic differentiation provides a signaling code that translates different combinations of extracellular signals into appropriate cellular responses. PMID- 12730291 TI - A tyrosine-based sorting signal is involved in connexin43 stability and gap junction turnover. AB - The gap junction protein connexin43 is known to have a rapid turnover, involving degradation by both the proteasomal and lysosomal systems, but the structural features of connexin43 that govern these actions are not known. The connexin43 C terminal sequence contains a proline-rich region corresponding to the consensus of a protein-protein interaction PY-motif (xPPxY), and an overlapping putative tyrosine-based sorting signal (Yxxphi; =hydrophobic), known to play a role in the intracellular trafficking of many membrane proteins. As both motifs may control turnover of connexin43, we used a combination of metabolic radiolabelling, immuno precipitation and functional assays to determine the possible role of these motifs in controlling degradation of human connexin43 expressed in SKHep1 cells. Mutation V289D in the tyrosine-based sorting motif increased the steady-state pool of connexin43 by approximately 3.5-fold, while mutation P283L in the PY motif produced a comparatively modest augmentation (1.7-fold). No additive effect was observed when the overlapping tyrosine was mutated. In pulse-chase experiments, the Y286A substitution increased the half-life of connexin43 from 2 to 6 hours, indicating that the increased steady-state levels reflected reduced protein degradation. Moreover, expression at the junctional membrane, as well as gap junction-mediated intercellular communication (GJC), were nearly abolished by lysosomal inhibitors and Brefeldin A in cells expressing wild-type connexin43, but were unaffected in the tyrosine mutant. These results provide strong evidence that the tyrosine-based motif of human connexin43 is a prime determinant controlling connexin43 stability, and consequently GJC, by targeting connexin43 for degradation in the endocytic/lysosomal compartment. PMID- 12730292 TI - Drosophila skpA, a component of SCF ubiquitin ligases, regulates centrosome duplication independently of cyclin E accumulation. AB - Centrosome duplication must be coupled to the main cell cycle to ensure that each cell has precisely two centrosomes at the onset of mitosis. Supernumerary centrosomes are commonly observed in cancer cells, and may contribute to tumorigenesis. Drosophila skpA, a component of SCF ubiquitin ligases, regulates the link between the cell and centrosome cycles. Lethal skpA null mutants exhibit dramatic centrosome overduplication and additional defects in chromatin condensation, cell cycle progression and endoreduplication. Surprisingly, many mutant cells are able to organize pseudo-bipolar spindles and execute a normal anaphase in the presence of extra functional centrosomes. SkpA mutant cells accumulate higher levels of cyclin E than wildtype cells during S and G2, suggesting that elevated cdk2/cyclin E activity may account for the supernumerary centrosomes in skpA- cells. However, centrosome overduplication still occurs in skpA-; cycE- mutant animals, demonstrating that high cyclin E levels are not necessary for centrosome overduplication. These data suggest that additional SCF targets regulate the centrosome duplication pathway. PMID- 12730293 TI - Cholesterol sulfate in human physiology: what's it all about? AB - Cholesterol sulfate is quantitatively the most important known sterol sulfate in human plasma, where it is present in a concentration that overlaps that of the other abundant circulating steroid sulfate, dehydroepiandrosterone (DHEA) sulfate. Although these sulfolipids have similar production and metabolic clearance rates, they arise from distinct sources and are metabolized by different pathways. While the function of DHEA sulfate remains an enigma, cholesterol sulfate has emerged as an important regulatory molecule. Cholesterol sulfate is a component of cell membranes where it has a stabilizing role, e.g., protecting erythrocytes from osmotic lysis and regulating sperm capacitation. It is present in platelet membranes where it supports platelet adhesion. Cholesterol sulfate can regulate the activity of serine proteases, e.g., those involved in blood clotting, fibrinolysis, and epidermal cell adhesion. As a result of its ability to regulate the activity of selective protein kinase C isoforms and modulate the specificity of phosphatidylinositol 3-kinase, cholesterol sulfate is involved in signal transduction. Cholesterol sulfate functions in keratinocyte differentiation, inducing genes that encode for key components involved in development of the barrier. The accumulating evidence demonstrating a regulatory function for cholesterol sulfate appears solid; the challenge now is to work out the molecular mechanisms whereby this interesting molecule carries out its various roles. PMID- 12730294 TI - Lack of genetic linkage evidence for a trans-acting factor having a large effect on plasma lipoprotein[a] levels in African Americans. AB - The distribution of plasma lipoprotein[a] (Lp[a]) concentrations, a risk factor for cardiovascular disease, varies greatly among racial groups, with African Americans having values that are shifted toward higher levels than those of whites. The underlying cause of this heterogeneity is unknown, but a role for "trans-acting" factors has been hypothesized. This study used genetic linkage analysis to localize genetic factors influencing Lp[a] levels in African Americans that were absent in other populations; linkage results were analyzed separately in non-Hispanic whites, Hispanic whites, and African Americans. As expected, all three samples showed highly significant linkage at the approximate location of the lysophosphatidic acid locus. The white populations also independently had regions of significant linkage on chromosome 19 (LOD 3.80) and suggestive linkage on chromosomes 12 (LOD 1.60), 14 (LOD 2.56), and 19 (LOD 2.52). No linkage evidence was found to support the hypothesis of another single gene with large effects specifically segregating in African Americans that may account for their elevated Lp[a] levels. PMID- 12730295 TI - Alterations of plasma lipids in mice via adenoviral-mediated hepatic overexpression of human ABCA1. AB - ATP binding cassette transporter A1 (ABCA1) is a widely expressed lipid transporter essential for the generation of HDL. ABCA1 is particularly abundant in the liver, suggesting that the liver may play a major role in HDL homeostasis. To determine how hepatic ABCA1 affects plasma HDL cholesterol levels, we treated mice with an adenovirus (Ad)-expressing human ABCA1 under the control of the cytomegalovirus promoter. Treated mice showed a dose-dependent increase in hepatic ABCA1 protein, ranging from 1.2-fold to 8.3-fold using doses from 5 x 108 to 1.5 x 109 pfu, with maximal expression observed on Day 3 posttreatment. A selective increase in HDL cholesterol occurred at Day 3 in mice treated with 5 x 108 pfu Ad-ABCA1, but higher doses did not further elevate HDL cholesterol levels. In contrast, total cholesterol, triglycerides, phospholipids, non-HDL cholesterol, and apolipoprotein B levels all increased in a dose-dependent manner, suggesting that excessive overexpression of hepatic ABCA1 in the absence of its normal regulatory sequences altered total lipid homeostasis. At comparable expression levels, bacterial artificial chromosome transgenic mice, which express ABCA1 under the control of its endogenous regulatory sequences, showed a greater and more specific increase in HDL cholesterol than Ad-ABCA1-treated mice. Our results suggest that appropriate regulation of ABCA1 is critical for a selective increase in HDL cholesterol levels. PMID- 12730297 TI - Plasma PAF-acetylhydrolase in patients with coronary artery disease: results of a cross-sectional analysis. AB - Inflammation underlies both onset and perpetuation of atherosclerosis. Plasma lipoproteins transport the platelet-activating factor-acetylhydrolase (PAF-AH) with potentially anti-inflammatory activities. Our aim was to determine whether PAF-AH activity was associated with inflammatory markers and with coronary artery disease (CAD). PAF-AH activity and a panel of inflammatory mediators were measured in plasma of 496 patients with CAD and in 477 controls; 276 patients presented with stable angina pectoris and 220 with acute coronary syndrome (ACS). Individuals within the highest quartile of PAF-AH activity had an 1.8-fold increase in CAD risk [95% confidence interval (CI), 1.01 to 3.2; P = 0.048] compared with those in the first quartile (adjusted for clinical and metabolic factors). When excluding individuals receiving statin and angiotensin-converting enzyme-inhibitor medication, individuals within the highest quartile of PAF-AH activity revealed a 3.9-fold increase in CAD risk (95% CI, 2.0 to 7.7; P < 0.0001). In these subjects, the plasma PAF-AH activity increased gradually in stable angina and in ACS both in men (P < 0.0001) and in women (P < 0.001), as compared with controls. No correlation was found between PAF-AH levels and those of common markers of inflammation. This study and the previous ones raise the important issue of whether PAF-AH is simply a marker of risk or directly promotes atherosclerosis. PMID- 12730296 TI - Phytosterols in low- and nonfat beverages as part of a controlled diet fail to lower plasma lipid levels. AB - Dietary phytosterols have been shown to reduce plasma cholesterol concentrations when consumed in different food matrices, but their effectiveness in nonfat or low-fat beverages has not been established. The objective of this study was to examine whether phytosterols alter plasma lipid levels when incorporated into nonfat or low-fat beverages. Fifteen moderately hypercholesterolemic men and women consumed three precisely controlled diets for periods of 21 days each in random order. Diets contained either a nonfat placebo beverage (NF), a beverage that is nonfat with added phytosterols (NFPS), or a beverage that is low in fat with added phytosterols (LFPS). Total cholesterol concentrations were not different between groups at endpoint, decreasing (P < 0.05) equally by 8.5%, 11.6%, and 10.1% with NF, NFPS, and LFPS consumption, respectively. There was no effect of dietary treatment on LDL cholesterol concentrations, which decreased over time (P < 0.05) by 5%, 10.4%, and 8.5% with NF, NFPS, and LFPS, respectively. HDL cholesterol and triacylglycerol concentrations were unaffected by the diets. Provision of phytosterols as part of nonfat and low-fat beverages did not exert any greater hypocholesterolemic effect than a nonfat placebo beverage. These results show that intake of phytosterols in a low-fat beverage format is not efficacious for lipid level modification. PMID- 12730298 TI - The surface cholesteryl ester content of donor and acceptor particles regulates CETP: a liposome-based approach to assess the substrate properties of lipoproteins. AB - Cholesteryl ester transfer protein (CETP) activity is regulated, in part, by lipoprotein composition. We previously demonstrated that CETP activity follows saturation kinetics as cholesteryl ester (CE) levels in the phospholipid surface of donor particles are increased. We propose here that the plateau of CETP activity occurs because the surface concentration of CE in the acceptor becomes rate limiting. This hypothesis was tested in CETP assays between synthetic liposomes whose CE content was varied independently. As donor CE increased, CETP activity followed saturable kinetics, but the slope of the first-order portion of the curve and the maximum achievable CE transfer rate were linearly related to the acceptor's surface CE concentration. These findings, plus studies with free cholesterol-modified LDL, strongly suggest that CE-rich donor liposomes can measure the CETP-accessible CE in acceptor lipoproteins. CETP activity from CE rich liposomes to multiple control LDLs ranged 1.8-fold despite equivalent CETP binding capacity, suggesting that LDLs vary widely in their capacity to present CE to CETP. Thus, CETP activity depends on the surface availability of substrate lipids in the donor and acceptor. Donor liposomes with high CE content can be used to assess how subtle changes in composition alter the substrate potential of plasma lipoproteins. PMID- 12730299 TI - Gamma-tocotrienol, a vitamin E homolog, is a natriuretic hormone precursor. AB - 2,7,8-Trimethyl-2-(beta-carboxyethyl)-6-hydroxychroman (gamma-CEHC), a metabolite of gamma-tocopherol and gamma-tocotrienol, was identified as a new endogenous natriuretic factor. However, gamma-tocopherol and gamma-tocotrienol, both precursors of gamma-CEHC, have never directly been observed to have natriuretic potency. Thus, we investigated whether gamma-tocotrienol could cause natriuresis and diuresis in rats. The rats were divided into two groups that were given a control or a high-sodium diet for 4 weeks, and then subdivided into placebo and gamma-tocotrienol subgroups given only corn oil-removed vitamin E and oil supplemented with gamma-tocotrienol, respectively. After oral administration of three experimental doses, rat urine was collected and gamma-CEHC, urine volume, sodium, and potassium content were determined. Only in rats given a high-NaCl diet did gamma-tocotrienol accelerate and increase sodium excretion, showing no effect on potassium excretion. Sodium excretion in the high-NaCl group given gamma-tocotrienol was 5.06 +/- 2.70 g/day, and in the control group given gamma tocotrienol, 0.11 +/- 0.06 g/day. Furthermore, gamma-tocotrienol affected urine volume in the specific condition of high-NaCl body stores and gamma-tocotrienol supplementation. In this study, we found that gamma-tocotrienol, one of the natural vitamin E homologs, stimulates sodium excretion in vivo, suggesting that gamma-tocotrienol possesses a hormone-like natriuretic function. PMID- 12730301 TI - LDL activates signaling pathways leading to an increase in cytosolic free calcium and stimulation of CD11b expression in monocytes. AB - In the present study, we investigated the mechanisms by which plasma lipoproteins modulate the integrin-dependent adhesion properties of monocytes. LDL induced the expression of the monocyte CD11b in vitro as well as in vivo via intracellular signaling mechanisms involving calcium transients. The effect on CD11b transcription was specific for native LDL and was blocked by a neutralizing anti LDL receptor antibody. Neither oxidized LDL nor HDL had any effect on CD11b expression. Although LDL stimulated CD11b surface expression, the integrins were not activated. To initiate the CD11b-specific adhesion to the endothelium, the engagement of chemokine receptor CCR2 and intact chemokine-to-integrin signaling was necessary. However, the activation of CCR2 with monocyte chemoattractant protein-1 not only stimulated the integrins preexisting on the cell surface, but also increased the number of CD11b molecules on the cell surface. This was particularly pronounced in THP-1 cells after treatment with LDL. In a previous study, we showed that LDL induces the expression of CCR2 in monocytes. We conclude that this may be the underlying cause of the enhanced chemokine effect on CD11b expression and activation observed with these cells. PMID- 12730302 TI - Regulation of human CETP gene expression: role of SP1 and SP3 transcription factors at promoter sites -690, -629, and -37. AB - Cholesteryl ester transfer protein (CETP) is a key factor in plasma reverse cholesterol transport and is implicated in the pathophysiology of atherogenic dyslipidemia. Variations observed in plasma CETP mass and activity in both normolipidemic and dyslipidemic individuals may reflect differences in CETP gene expression. We evaluated the respective roles of the Sp1 and Sp3 transcription factors on the promoter activity of the human CETP gene at a new Sp1/Sp3 site identified at position -690, and at two previously described Sp1/Sp3 sites at positions -37 and -629. In transient transfection in HepG2 cells, site-directed mutagenesis using luciferase reporter constructs containing a promoter fragment from +32 to -745 indicated that the new -690 site acts as a repressive element in reducing CETP promoter activity (-22%; P < 0.05); equally, this site exerts an additive effect with the -629 site, inducing marked repression (-42%; P < 0.005). In contrast, in NCTC cells that display a 16-fold lower level of Sp3, the repressive effect at the -690 site was enhanced 2-fold (-45%; P < 0.05), whereas the -629 site exerted no effect. Cotransfection of Sp1 and/or Sp3 in SL2 insect cells lacking endogenous Sp factors demonstrated that Sp1 and Sp3 act as activators at the -690 and -37 sites, whereas Sp3 acts as a repressor at the -629 site. Taken together, our data demonstrate that Sp1 and Sp3 regulate human CETP promoter activity through three Sp1/Sp3 binding sites in a distinct manner, and that the Sp1/Sp3 ratio is a key factor in determining the relative contribution of these sites to total promoter activity. PMID- 12730300 TI - Isomer-specific regulation of metabolism and PPARgamma signaling by CLA in human preadipocytes. AB - Trans-10,cis-12 conjugated linoleic acid (CLA) has previously been shown to be the CLA isomer responsible for CLA-induced reductions in body fat in animal models, and we have shown that this isomer, but not the cis-9,trans-11 CLA isomer, specifically decreased triglyceride (TG) accumulation in primary human adipocytes in vitro. Here we investigated the mechanism behind the isomer specific, CLA-mediated reduction in TG accumulation in differentiating human preadipocytes. Trans-10,cis-12 CLA decreased insulin-stimulated glucose uptake and oxidation, and reduced insulin-dependent glucose transporter 4 gene expression. Furthermore, trans-10,cis-12 CLA reduced oleic acid uptake and oxidation when compared with all other treatments. In parallel to CLA's effects on metabolism, trans-10,cis-12 CLA decreased, whereas cis-9,trans-11 CLA increased, the expression of peroxisome proliferator-activated receptor gamma (PPARgamma) and several of its downstream target genes when compared with vehicle controls. Transient transfections demonstrated that both CLA isomers antagonized ligand-dependent activation of PPARgamma. Collectively, trans-10,cis-12, but not cis-9, trans-11, CLA decreased glucose and lipid uptake and oxidation and preadipocyte differentiation by altering preadipocyte gene transcription in a manner that appeared to be due, in part, to decreased PPARgamma expression. PMID- 12730303 TI - LDL immune complexes stimulate LDL receptor expression in U937 histiocytes via extracellular signal-regulated kinase and AP-1. AB - We have previously shown that LDL-containing immune complexes (LDL-ICs) induce up regulation of LDL receptor (LDLR) expression in human macrophages. The present study further investigated the molecular mechanisms leading to LDLR up-regulation by LDL-ICs as well as the signaling pathways involved. Results showed that treatment of U937 histiocytes with LDL-ICs did not increase the precursors and the cleaved forms of sterol-regulatory element binding proteins (SREBPs) 1a and 2, suggesting that SREBPs may not be involved in LDLR up-regulation by LDL-ICs. Promoter deletion and mutation studies showed that the AP-1 binding sites were essential for LDL-IC-stimulated LDLR expression. Electrophoretic mobility shift assays further demonstrated that LDL-ICs stimulated transcription factor AP-1 activity. Studies assessing the signaling pathways involved in LDLR up-regulation by LDL-ICs showed that the up-regulation of LDLR was extracellular signal regulated kinase (ERK) dependent. In conclusion, the present study shows that LDL ICs up-regulate LDLR expression via the ERK signaling pathway and the AP-1 motif dependent transcriptional activation. PMID- 12730305 TI - Simultaneous quantification of lyso-neutral glycosphingolipids and neutral glycosphingolipids by N-acetylation with [3H]acetic anhydride. AB - We describe a new method that permits quantification in the pmol to nmol range of three lyso-neutral glycosphingolipids (lyso-n-GSLs), glucosylsphingosine (GlcSph), galactosylsphingosine (GalSph), and lactosylsphingosine, in the same sample as neutral glycosphingolipids (n-GSLs). Lyso-n-GSLs and n-GSLs are initially obtained from a crude lipid extract using Sephadex G25 chromatography, followed by their isolation in one fraction, which is devoid of other contaminating lipids, by aminopropyl solid-phase chromatography. Lyso-n-GSLs and n-GSLs are subsequently separated from one another by weak cation exchange chromatography. N-GSLs are then deacylated by strong alkaline hydrolysis, and the N-deacylated-GSLs and lyso-n-GSLs are subsequently N-acetylated using [3H]acetic anhydride. An optimal concentration of 5 mM acetic anhydride was established, which gave >95% N-acetylation. We demonstrate the usefulness of this technique by showing an approximately 40-fold increase of both GlcSph and glucosylceramide in brain tissue from a glucocerebrosidase-deficient mouse, as well as significant lactosylceramide accumulation. The application and optimization of this technique for lyso-n-GSLs and lyso-GSLs will permit their quantification in small amounts of biological tissues, particularly in the GSL storage diseases, such as Gaucher and Krabbe's disease, in which GlcSph and GalSph, respectively, accumulate. PMID- 12730304 TI - Phospholipid transfer protein is present in human atherosclerotic lesions and is expressed by macrophages and foam cells. AB - Phospholipid transfer protein (PLTP) in plasma promotes phospholipid transfer from triglyceride-rich lipoproteins to HDL and plays a major role in HDL remodeling. Recent in vivo observations also support a key role for PLTP in cholesterol metabolism. Our immunohistochemical analysis of human carotid endarterectomy samples identified immunoreactive PLTP in areas that colocalized with CD68-positive macrophages, suggesting that PLTP could be produced locally by intimal macrophages. Using RT-PCR, Western blot analysis with a monoclonal anti PLTP antibody, and a PLTP activity assay, we observed PLTP mRNA and protein expression in human macrophages. In adherent peripheral blood human macrophages, this PLTP expression was increased by culture with granulocyte macrophage colony stimulating factor. Incubation of macrophages with acetylated-LDL induced an increase in PLTP mRNA and protein expression that paralleled cholesterol loading. PLTP expression was observed in elicited mouse peritoneal macrophages and in cultured Raw264.7 cells as well. Thus, this study demonstrates that PLTP is expressed by macrophages, is regulated by cholesterol loading, and is present in atherosclerotic lesions. PMID- 12730307 TI - Positive and negative affect in very old age. AB - The current study examined two issues involving the relationship between age and affect in very old age using data from men and women (aged 70 to 100+ years, M = 85 years) in the Berlin Aging Study (BASE). The first issue was whether unique effects of age on positive and negative affect remained after we controlled for other variables that would be expected to relate to affect in late life. We found no unique effects of age after we controlled for demographic, personality, and health and cognitive functioning variables. Personality and general intelligence emerged as the strongest predictors of positive and negative affect. Second, we evaluated patterns within meaningful subgroups: young old versus oldest old and men versus women. Subgroup differences in predictor patterns were minimal. Although we accounted for much of the age-related variance in positive and negative affect, a significant amount of variance in the affect of older adults remained unexplained. PMID- 12730306 TI - Measurement of cholesterol of major serum lipoprotein classes by anion-exchange HPLC with perchlorate ion-containing eluent. AB - We have developed a high-performance liquid chromatography (HPLC) method for measurement of cholesterol in the major classes of serum lipoproteins, i.e., HDL, LDL, IDL, VLDL, and chylomicrons. Lipoproteins in serum were separated on a column containing diethylaminoethyl-ligand nonporous polymer-based gel by elution with a step gradient of sodium perchlorate concentration, and detected by post column reaction with a reagent containing cholesterol esterase and cholesterol oxidase. The within-day assay and between-day assay coefficients of variation for cholesterol concentration in lipoproteins were in the ranges of 0.9-6.4% and 1.1 11.9%, respectively. The correlation coefficients between the values of HDL, LDL, IDL, VLDL, and chylomicron cholesterol measured by the HPLC method and those estimated by an ultracentrifugation method were 0.892, 0.921, 0.840, 0.930, and 0.873, respectively. Values of remnant-like particle cholesterol measured by an immunoseparation technique (Japan Immunoresearch Laboratories, Japan) were significantly correlated with VLDL and chylomicron cholesterol values measured by the HPLC method (r = 0.883 and r = 0.729, respectively). This rapid and accurate HPLC method was successfully applied to the analysis of plasma lipoproteins of patients with hyperlipidemia. PMID- 12730309 TI - Stability and change in adult personality over 6 years: findings from the Victoria Longitudinal Study. AB - Data from the Victoria Longitudinal Study were used to examine the 6-year longitudinal stability of personality in older adults. Personality was measured with the NEO Personality Inventory. The longitudinal sample consisted of 223 adults initially ranging from 55 to 85 years of age. Longitudinal confirmatory factor analyses were used to examine the stability of individual differences in change over time, and the stability of the longitudinal factor structure. The results indicated both substantial stability at the level of individual differences in change, as well as significant individual differences in change that were related to age and gender. Finally, the factor structure of personality was invariant over time but did not approximate simple structure for the five dimensions of personality. Our study of 6-year personality development provided both (a). a confirmation of early significant stability findings and (b). unique evidence for significant individual differences in late adulthood. PMID- 12730310 TI - Application of the five-factor model of personality to intergenerational perception. AB - The perception held by respondents in a full range of age groups (young, middle aged, and old) toward a full range of target age groups (young, middle-aged, and old) was studied in a multidimensional way. The five-factor model of personality was chosen as the model of intergenerational perception and the Gough Adjective Checklist was chosen as the measurement device. A total of 867 participants rated the degree to which they thought 300 different adjectives described people of various ages. Responses were factor analyzed, and the five expected factors were found. For Conscientiousness, the factor scores showed a linearly increasing pattern, with young children and children scoring lower, and the middle-aged, adults, elderly people, and the very old scoring higher. For Openness, the reverse trend was observed, but the real decrease started at the young adult age. For Neuroticism, factor scores were very low when the targets were children, very high when the targets were adolescents, and neither high nor low when the targets were middle-aged, elderly, or very old. For Introversion, the pattern of scores appeared U shaped, with adolescents and young and middle-aged adults on one side and the other targets on the opposite side. Finally, for Agreeableness, the middle-aged adults scored much lower than all the other targets. Overall, the age of the target effect explained on the average more than two thirds of the explained variance, whereas the age of the respondent variable explained less than one tenth of the explained variance. It appears that, in the population, a strong consensus is held concerning the attribution of personality characteristics to various age groups. PMID- 12730308 TI - Modeling intraindividual change in personality traits: findings from the normative aging study. AB - To advance an intraindividual life-span approach to the issue of stability and change, we studied personality trait trajectories in adulthood. Growth curves for extraversion and neuroticism were estimated for over 1600 men (initially aged 43 91) in the Normative Aging Study, who were followed over 12 years. We found significant individual differences in intraindividual change for both traits, as well as different trajectories for extraversion and neuroticism. The overall extraversion trajectory was best defined by a linear model, but neuroticism was characterized by quadratic decline with age. We then considered several variables as predictors of individual differences around these overall trajectories. Birth cohort, marriage or remarriage, death of spouse, and memory complaints were all significant predictors, explaining variability in both level and rate of personality trait change. These findings suggest that there is a good deal of variability in personality trajectories, and that some of this variability can be explained by birth cohort as well as by age-graded life events. PMID- 12730311 TI - Age-related slowing of digit symbol substitution revisited: what do longitudinal age changes reflect? AB - A previous investigation reported that cross-sectional age differences in Digit Symbol Substitution (DSS) test performance reflect declines in perceptual processing speed. Support for the tenability of the processing speed hypothesis requires examining whether longitudinal age-related change in DSS performance is largely mediated by changes in speed. The present study used data from the Victoria Longitudinal Study to examine patterns and predictors of longitudinal change in DSS for 512 older adults (M(age) = 68.37 years, SD = 7.43). On the basis of multilevel modeling, baseline DSS performance was poorer for older participants and men, with longitudinal declines more pronounced with increasing age and decreasing speed. In contrast to the present cross-sectional findings, statistical control of change trajectories in perceptual speed using the same data did not substantially attenuate age changes. These discrepancies suggest different sources of variance may underlie cross-sectional age differences and longitudinal age changes for DSS. PMID- 12730312 TI - Give day care a chance to be effective: a commentary. PMID- 12730314 TI - Effects of volunteering on the well-being of older adults. AB - OBJECTIVES: This study tests the effects of volunteering on the well-being of older adults, including the effect of level of engagement, the moderating effects of demographic and social factors, and the effects of the nature of the volunteer experience. METHODS: This is a secondary data analysis of three waves of data from the Americans' Changing Lives Study. Self-rated health, functional dependency, and depression are regressed on the well-being measures from the previous waves, other control variables and volunteer status, volunteer hours, type and number of volunteer organizations, and the perceived benefit to others of the volunteer work. RESULTS: Older adults who volunteer and who engage in more hours of volunteering report higher levels of well-being. This positive effect was not moderated by social integration, race, or gender. There was no effect of the number of organizations for which the older adult volunteered, the type of organization, or the perceived benefit of the work to others. DISCUSSION: This work contributes to a knowledge base that points to the development of social programs and policies that maximize the engagement of older adults in volunteer roles. The findings suggest that targeting efforts may not be warranted, in that there are not differential benefits according to personal characteristics of the volunteer. Future studies have to address the nature of the social institutions that will maximize the number of elders in these roles and the benefits that they accrue. PMID- 12730315 TI - 9-11, personal stress, mental health, and sense of control among older adults. AB - OBJECTIVES: We assessed whether the events of 9-11 affected the personal stress, mental health, or sense of control of older adults participating in an ongoing longitudinal study, and whether baseline characteristics were associated with the magnitude of any such changes. METHODS: Personal stress, mental health, and sense of control were measured at baseline and at six bimonthly follow-up interviews among 1662 patients. Of these, 437 had the opportunity to complete three interviews before and after 9-11, with 291 (67%) completing all six. We performed graphic comparisons, paired t tests, classification based on standard errors of measurement (SEMs), and multiple linear regressions for patients who completed all six interviews. RESULTS: No noticeable changes in aggregate trends for personal stress or mental health were associated with 9-11. However, 9-11 was associated with an aggregate decline in sense of control. This decline in sense of control was greater among those who were working for pay, had more comfortable incomes, and reported greater religiosity. DISCUSSION: Older adults more closely resembling those who died during 9-11 and those with greater levels of religiosity were most likely to have their sense of control affected by this catastrophic event. PMID- 12730316 TI - Creating continuity through mutual assistance: intergenerational reciprocity in four ethnic groups. AB - OBJECTIVES: Our purpose is to examine how culture-specific conceptions of mutual assistance in four ethnic groups reflect the nature of social exchange and its role in creating continuity. METHODS: We conducted five in-depth interviews over a 5-year period with 270 respondents aged 50 and up (59 African Americans, 85 Latinos, 78 Filipino Americans, and 48 Cambodian Americans). We asked both open ended and semistructured questions to determine how respondents viewed their role in mutual assistance in the family. RESULTS: Mutual assistance was a critical element in intergenerational relations in all four groups, as it represented continuity within the family and was a part of a broader cultural ethos of family relationships. There were differences between groups in overall approach to mutual assistance, the factors to which they assigned the greatest importance, and the degree of dissatisfaction expressed over family relationships. DISCUSSION: Social exchange can be viewed not only as a means by which those who are old maintain power but as a major vehicle for perpetuating continuity across the generations. When social exchange breaks down, elders suffer from loss of power as expressed through role loss. Elders must have a negotiable commodity to exchange as well as the flexibility to adjust to changes in the extended family. Maintaining continuity of the family in the face of disruptive threats poses a particular challenge for elders. PMID- 12730317 TI - Religious meaning and subjective well-being in late life. AB - OBJECTIVES: The purpose of this study is to examine the relationship between religious meaning and subjective well-being. A major emphasis is placed on assessing race differences in the relationship between these constructs. METHODS: Interviews were conducted with a nationwide sample of older White and older Black adults. Survey items were administered to assess a sense of meaning in life that is derived specifically from religion. Subjective well-being was measured with indices of life satisfaction, self-esteem, and optimism. RESULTS: The findings suggest that older adults who derive a sense of meaning in life from religion tend to have higher levels of life satisfaction, self-esteem, and optimism. The data further reveal that older Black adults are more likely to find meaning in religion than older White adults. In addition, the relationships among religious meaning, life satisfaction, self-esteem, and optimism tend to be stronger for older African Americans persons than older White persons. DISCUSSION: Researchers have argued for some time that religion may be an important source of resilience for older Black adults, but it is not clear how these beneficial effects arise. The data from this study suggest that religious meaning may be an important factor. PMID- 12730318 TI - The Effect of altering ADL thresholds on active life expectancy estimates for older persons. AB - OBJECTIVES: Research on disability and active life expectancy (ALE) has often criticized the measurement of disability but has rarely empirically investigated the effect of changing measurement. The purpose of this study was to determine whether altering the number of activities of daily living (ADLs) required to consider an individual "disabled" affects population-based ALE estimates after considering parametric uncertainty and sampling error. METHODS: The authors develop a Bayesian approach to estimating multistate life tables for a three dimensional state space, using data on community-dwelling older adults from the 1989 and 1994 National Long Term Care Survey analytic files. Empirical confidence intervals for ALE are compared across 6 models using successively higher ADL cutoffs for defining individuals as being disabled. RESULTS: After considering sampling and other errors in the estimation of transition probabilities, the authors found that altering the threshold for measuring disability has relatively little effect on ALE estimates, especially with higher ADL-level thresholds and at older ages. DISCUSSION: The implications of the results include that disability measurement, including altering the definition of being disabled and possibly expanding the state space of a model, may not affect population-based estimates of ALE. PMID- 12730319 TI - Life expectancy with cognitive impairment in the older population of the United States. AB - OBJECTIVES: This article provides estimates of the prevalence of cognitive impairment by age and sex for a nationally representative sample of the U.S. population aged 70 and over. From these estimates, years of life with and without cognitive impairment are calculated. METHODS: Using data from the Assets and Health Dynamics of the Oldest Old (AHEAD) survey, the prevalence of cognitive impairment is estimated for a sample representing both the community-dwelling and institutionalized older American population. Sullivan's method is used to calculate the average number of years an elderly person can expect to live with and without cognitive impairment. RESULTS: The prevalence of moderate to severe cognitive impairment in the total U.S. population aged 70 and over is 9.5%. At age 70, the average American can expect 1.5 years with cognitive impairment. Expected length of life with cognitive impairment is longer for women than men because of their longer life expectancy. DISCUSSION: As total life expectancy continues to increase, the length of life with cognitive impairment for the American population will increase unless age-specific prevalence is reduced. There is great potential for improvement in future early treatment and diagnosis of this condition. PMID- 12730320 TI - Predicting 12-month mortality for persons with dementia. AB - OBJECTIVES: We develop and test a model of 12-month mortality among persons (N=3858) with organic dementia. METHODS: Data are from caregiver interviews and claims records for persons enrolled in the Medicare Alzheimer's Disease Demonstration Evaluation. Information covers the year prior to enrollment through 36 months. We used Proportional hazards models to predict time to death. We estimated two starting points: first, the date of randomization, and second, the date of maximum difficulty in performing two or more activities of daily living (ADLs). RESULTS: The final model includes age, gender, ADL difficulty, medical conditions, prior year hospitalizations, and whether a daughter was the primary caregiver. We combined hazard ratios to produce a cumulative mortality risk score. Model discrimination is reasonable for both models (c statistics of.72 and.69, respectively), and calibration tests were nonsignificant. DISCUSSION: The model's efficiency, as measured by the ratio of false positives (those predicted to die, but who lived) to true positives (those predicted to die and who did die) ranged from fewer than 1:1 to more than 4:1 as the model's sensitivity increased. This ratio was lower in the two or more ADL difficulty model. A validation test of the prediction model found comparable sensitivity and specificity (c statistic of.69) to the reference model. PMID- 12730321 TI - Common endocrine control of body weight, reproduction, and bone mass. AB - Bone mass is maintained constant between puberty and menopause by the balance between osteoblast and osteoclast activity. The existence of a hormonal control of osteoblast activity has been speculated for years by analogy to osteoclast biology. Through the search for such humoral signal(s) regulating bone formation, leptin has been identified as a strong inhibitor of bone formation. Furthermore, intracerebroventricular infusion of leptin has shown that the effect of this adipocyte-derived hormone on bone is mediated via a brain relay. Subsequent studies have led to the identification of hypothalamic groups of neurons involved in leptin's antiosteogenic function. In addition, those neurons or neuronal pathways are distinct from neurons responsible for the regulation of energy metabolism. Finally, the peripheral mediator of leptin's antiosteogenic function has been identified as the sympathetic nervous system. Sympathomimetics administered to mice decreased bone formation and bone mass. Conversely, beta blockers increased bone formation and bone mass and blunted the bone loss induced by ovariectomy. PMID- 12730322 TI - Notch and Presenilin: regulated intramembrane proteolysis links development and degeneration. AB - Intensive studies of three proteins--Presenilin, Notch, and the amyloid precursor protein (APP)--have led to the recognition of a direct intersection between early development and late-life neurodegeneration. Notch signaling mediates many different intercellular communication events that are essential for determining the fates of neural and nonneural cells during development and in the adult. The Notch receptor acts in a core pathway as a membrane-bound transcription factor that is released to the nucleus by a two-step cleavage mechanism called regulated intramembrane proteolysis (RIP). The second cleavage is effected by Presenilin, an unusual polytopic aspartyl protease that apparently cleaves Notch and numerous other single-transmembrane substrates within the lipid bilayer. Another Presenilin substrate, APP, releases the amyloid ss-protein that can accumulate over time in limbic and association cortices and help initiate Alzheimer's disease. Elucidating the detailed mechanism of Presenilin processing of membrane proteins is important for understanding diverse signal transduction pathways and potentially for treating and preventing Alzheimer's disease. PMID- 12730323 TI - Brucella stationary-phase gene expression and virulence. AB - The capacity of the Brucella spp. to establish and maintain long-term residence in the phagosomal compartment of host macrophages is critical to their ability to produce chronic infections in their mammalian hosts. The RNA binding protein host factor I (HF-I) encoded by the hfq gene is required for the efficient translation of the stationary-phase sigma factor RpoS in many bacteria, and a Brucella abortus hfq mutant displays a phenotype in vitro, which suggests that it has a generalized defect in stationary-phase physiology. The inability of the B. abortus hfq mutant to survive and replicate in a wild-type manner in cultured murine macrophages, and the profound attenuation displayed by this strain and its B. melitensis counterpart in experimentally infected animals indicate that stationary-phase physiology plays an essential role in the capacity of the brucellae to establish and maintain long-term intracellular residence in host macrophages. The nature of the Brucella HF-I-regulated genes that have been identified to date suggests that the corresponding gene products contribute to the remarkable capacity of the brucellae to resist the harsh environmental conditions they encounter during their prolonged residence in the phagosomal compartment. PMID- 12730324 TI - Nitrogen assimilation and global regulation in Escherichia coli. AB - Nitrogen limitation in Escherichia coli controls the expression of about 100 genes of the nitrogen regulated (Ntr) response, including the ammonia assimilating glutamine synthetase. Low intracellular glutamine controls the Ntr response through several regulators, whose activities are modulated by a variety of metabolites. Ntr proteins assimilate ammonia, scavenge nitrogen-containing compounds, and appear to integrate ammonia assimilation with other aspects of metabolism, such as polyamine metabolism and glutamate synthesis. The leucine responsive regulatory protein (Lrp) controls the synthesis of glutamate synthase, which controls the Ntr response, presumably through its effect on intracellular glutamine. Some Ntr proteins inhibit the expression of some Lrp-activated genes. Guanosine tetraphosphate appears to control Lrp synthesis. In summary, a network of interacting global regulators that senses different aspects of metabolism integrates nitrogen assimilation with other metabolic processes. PMID- 12730325 TI - How bacteria assemble flagella. AB - The bacterial flagellum is both a motor organelle and a protein export/assembly apparatus. It extends from the cytoplasm to the cell exterior. All the protein subunits of the external elements have to be exported. Export employs a type III pathway, also utilized for secretion of virulence factors. Six of the components of the export apparatus are integral membrane proteins and are believed to be located within the flagellar basal body. Three others are soluble: the ATPase that drives export, a regulator of the ATPase, and a general chaperone. Exported substrates diffuse down a narrow channel in the growing structure and assemble at the distal end, often with the help of a capping structure. PMID- 12730326 TI - A salvage pathway for protein structures: tmRNA and trans-translation. AB - Transfer-messenger RNA (tmRNA, or SsrA), found in all eubacteria, has both transfer and messenger RNA activity. Relieving ribosome stalling by a process called trans-translation, tmRNAala enters the ribosome and adds its aminoacylated alanine to the nascent polypeptide. The original mRNA is released and tmRNA becomes the template for translation of a 10-amino-acid tag that signals for proteolytic degradation. Although essential in a few bacterial species, tmRNA is nonessential in Escherichia coli and many other bacteria. Proteins known to be associated with tmRNA include SmpB, ribosomal protein S1, RNase R, and phosphoribosyl pyrophosphate. SmpB, having no other known function, is essential for tmRNA activity. trans-translation operates within ribosomes stalled both at the end of truncated mRNAs and at rare codons and some natural termination sites. Both the release of stalled ribosomes and the subsequent degradation of tagged proteins are important consequences of trans-translation. PMID- 12730327 TI - Dominant-negative nuclear receptor corepressor relieves transcriptional inhibition of retinoic acid receptor but does not alter the agonist/antagonist activities of the tamoxifen-bound estrogen receptor. AB - Repression of the transcriptional activities of the estrogen receptor (ER) is a main goal in the treatment of breast cancer. The antiestrogen tamoxifen is an effective therapy for breast cancer patients because it inhibits estrogen stimulated gene expression and cell proliferation. Previous studies have implicated a complex containing the nuclear receptor corepressor (N-CoR) in the mechanism by which tamoxifen represses ER-mediated transcriptional activity. In the present study a truncated N-CoR construct was used to inhibit endogenous N CoR activity in an ER-positive breast cancer cell line. This dominant-negative N CoR was successful in relieving repression conferred by the unliganded retinoic acid receptor, but it failed to affect the transcriptional activity of the ER in the presence of tamoxifen. Correspondingly, the histone acetylation levels of nucleosomes on endogenous estrogen-responsive genes were unaltered in cells expressing the N-CoR dominant-negative, regardless of ligand. In addition, in vitro cell proliferation and in vivo tumor growth were unchanged in cells that express dominant-negative N-CoR. In conclusion, these results may reveal that N CoR affects tamoxifen-liganded ER in a manner distinct from its influence on retinoic acid receptor-mediated transcriptional activity or that corepressors other than N-CoR may be involved in the ability of tamoxifen to repress estrogen responsive transcription and tumor growth. PMID- 12730329 TI - Lysophosphatidic acid signals through mitogen-activated protein kinase extracellular signal regulated kinase in ovarian theca cells expressing the LPA1/edg2-receptor: involvement of a nonclassical pathway? AB - We investigated the mechanism of lysophosphatidic acid (LPA) signaling in ovarian theca cells and observed that stimulation with this bioactive lipid markedly enhanced Thr/Tyr phosphorylation of the MAPK ERK1/2. Activation of ERK was transient, showing a peak at 5 min that declined thereafter, and was not associated with a concomitant nuclear translocation of the enzyme, suggesting that a cytosolic tyrosine phosphatase may be responsible for switching off the signal. Epidermal growth factor (EGF)-induced activation of the enzyme in the same cell system was more rapid (peaking at 1 min), sustainable for at least 60 min, and could be suppressed by prior treatment with either pertussis toxin or a noncompetitive inhibitor of Ras acceptor protein, manumycin A. This functional inhibition of either Gi or Ras failed, however, to affect the LPA-induced ERK phosphorylation. Surprisingly, functional inhibition of Rho-GTPase, in C3 exotoxin-lipofected cells, markedly reduced LPA-stimulated phosphorylation of ERK, without affecting the EGF-induced stimulation of MAPK. Theca cells labeled with anti-LPA1/edg2-type antibody showed a distinct cell surface labeling, which is reflected in the expression of (LPA1)-type LPA receptors at both mRNA and protein levels. The findings indicate that LPA transiently stimulates MAPK ERK in LPA1/edg2-expressing theca cells and suggest an alternative mechanism regulating the activation of ERK that differs from the canonical EGF-Ras-MAPK kinase pathway. PMID- 12730328 TI - Nuclear factor Y and steroidogenic factor 1 physically and functionally interact to contribute to cell-specific expression of the mouse Follicle-stimulating hormone-beta gene. AB - FSH is a heterodimeric glycoprotein hormone secreted from the gonadotrope cell population of the anterior pituitary. Despite its crucial role in mammalian reproduction, very little is known about regulation of the FSH beta-subunit gene at the molecular level. In this report, we examine the basis for cell-specific expression of FSH beta using the mouse L beta T2 and alpha T3-1 gonadotrope derived cell lines. Characterization of the hormonal content of L beta T2 and alpha T3-1 cells at the protein level classifies these cells as relatively mature and immature gonadotropes, respectively. We studied L beta T2 cell-specific expression of FSH beta using 398 bp of the mouse FSH beta regulatory region linked to a luciferase reporter gene in transient transfection assays. This mouse FSH beta promoter can direct reporter gene expression specifically to L beta T2 cells when compared with other pituitary- and non-pituitary-derived cell lines, including alpha T3-1 cells. Furthermore, it is induced by activin, and interruption of the autocrine activin loop in L beta T2 cells by the addition of follistatin reduces its expression. Truncation analysis indicates that several regions of the promoter are involved in this specificity and that these can be dissociated from activin regulation. We identify binding sites for the orphan nuclear receptor steroidogenic factor-1 and the heterotrimeric transcription factor nuclear factor Y and show that these elements functionally interact to regulate FSH beta gene expression in an L beta T2 cell-specific manner. Moreover, steroidogenic factor-1 and nuclear factor Y are shown to physically interact with each other. This study is the first to demonstrate the presence of basal FSH beta protein in L beta T2 cells and to identify specific elements within the FSH beta promoter that contribute to basal and cell-specific expression of the gene. PMID- 12730330 TI - Functional characterization of the human resistin promoter with adipocyte determination- and differentiation-dependent factor 1/sterol regulatory element binding protein 1c and CCAAT enhancer binding protein-alpha. AB - Recent studies with murine models propose that resistin would be a possible mediator to link between obesity and insulin resistance. Although it has been reported that resistin is highly expressed and secreted by adipocytes, transcription factors that are involved in resistin gene expression have not been well characterized. To investigate the molecular mechanisms of resistin gene expression, we cloned and characterized the human resistin promoter. Sequence analysis of the resistin promoter revealed several putative binding sites for adipogenic transcription factors including adipocyte determination- and differentiation-dependent factor 1 (ADD1)/sterol regulatory element binding protein 1c (SREBP1c) and CCAAT enhancer binding protein-alpha (C/EBP alpha). EMSA and chromatin immunoprecipitation assays demonstrated that ADD1/SREBP1c binds to the human resistin promoter in vitro and in vivo. Expression of ADD1/SREBP1c transactivated the luciferase reporter gene activity, the promoter region of which contains a human resistin promoter in a sterol regulatory element (SRE) dependent manner. Furthermore, ectopic expression of ADD1/SREBP1c by adenovirus significantly increased the expression of resistin mRNA in adipocytes. Human resistin promoter was also activated by C/EBP alpha expression, although ectopic expression of both transcription factors did not show any synergistic effects on the activation of resistin promoter. Together, these data suggest that ADD1/SREBP1c and C/EBP alpha may play discrete roles in the regulation of the resistin gene expression. PMID- 12730331 TI - Cross-talk between peroxisome proliferator-activated receptor (PPAR) alpha and liver X receptor (LXR) in nutritional regulation of fatty acid metabolism. I. PPARs suppress sterol regulatory element binding protein-1c promoter through inhibition of LXR signaling. AB - Liver X receptors (LXRs) and peroxisome proliferator-activated receptors (PPARs) are members of nuclear receptors that form obligate heterodimers with retinoid X receptors (RXRs). These nuclear receptors play crucial roles in the regulation of fatty acid metabolism: LXRs activate expression of sterol regulatory element binding protein 1c (SREBP-1c), a dominant lipogenic gene regulator, whereas PPARalpha promotes fatty acid beta-oxidation genes. In the current study, effects of PPARs on the LXR-SREBP-1c pathway were investigated. Luciferase assays in human embryonic kidney 293 cells showed that overexpression of PPARalpha and gamma dose-dependently inhibited SREBP-1c promoter activity induced by LXR. Deletion and mutation studies demonstrated that the two LXR response elements (LXREs) in the SREBP-1c promoter region are responsible for this inhibitory effect of PPARs. Gel shift assays indicated that PPARs reduce binding of LXR/RXR to LXRE. PPARalpha-selective agonist enhanced these inhibitory effects. Supplementation with RXR attenuated these inhibitions by PPARs in luciferase and gel shift assays, implicating receptor interaction among LXR, PPAR, and RXR as a plausible mechanism. Competition of PPARalpha ligand with LXR ligand was observed in LXR/RXR binding to LXRE in gel shift assay, in LXR/RXR formation in nuclear extracts by coimmunoprecipitation, and in gene expression of SREBP-1c by Northern blot analysis of rat primary hepatocytes and mouse liver RNA. These data suggest that PPARalpha activation can suppress LXR-SREBP-1c pathway through reduction of LXR/RXR formation, proposing a novel transcription factor cross-talk between LXR and PPARalpha in hepatic lipid homeostasis. PMID- 12730333 TI - Baa, Baa, black sheep, are your kidneys full? PMID- 12730332 TI - Cross-talk between peroxisome proliferator-activated receptor (PPAR) alpha and liver X receptor (LXR) in nutritional regulation of fatty acid metabolism. II. LXRs suppress lipid degradation gene promoters through inhibition of PPAR signaling. AB - Fatty acid metabolism is transcriptionally regulated by two reciprocal systems: peroxisome proliferator-activated receptor (PPAR) alpha controls fatty acid degradation, whereas sterol regulatory element-binding protein-1c activated by liver X receptor (LXR) regulates fatty acid synthesis. To explore potential interactions between LXR and PPAR, the effect of LXR activation on PPARalpha signaling was investigated. In luciferase reporter gene assays, overexpression of LXRalpha or beta suppressed PPARalpha-induced peroxisome proliferator response element-luciferase activity in a dose-dependent manner. LXR agonists, T0901317 and 22(R)-hydroxycholesterol, dose dependently enhanced the suppressive effects of LXRs. Gel shift assays demonstrated that LXR reduced binding of PPARalpha/retinoid X receptor (RXR) alpha to peroxisome proliferator response element. Addition of increasing amounts of RXRalpha restored these inhibitory effects in both luciferase and gel shift assays, suggesting the presence of RXRalpha competition. In vitro protein binding assays demonstrated that activation of LXR by an LXR agonist promoted formation of LXR/RXRalpha and, more importantly, LXR/PPARalpha heterodimers, leading to a reduction of PPARalpha/RXRalpha formation. Supportively, in vivo administration of the LXR ligand to mice and rat primary hepatocytes substantially decreased hepatic mRNA levels of PPARalpha-targeted genes in both basal and PPARalpha agonist-induced conditions. The amount of nuclear PPARalpha/RXR heterodimers in the mouse livers was induced by treatment with PPARalpha ligand, and was suppressed by superimposed LXR ligand. Taken together with data from the accompanying paper (Yoshikawa, T., T. Ide, H. Shimano, N. Yahagi, M. Amemiya-Kudo, T. Matsuzaka, S. Yatoh, T. Kitamine, H. Okazaki, Y. Tamura, M. Sekiya, A. Takahashi, A. H. Hasty, R. Sato, H. Sone, J. Osuga, S. Ishibashi, and N. Yamada, Endocrinology 144:1240 1254) describing PPARalpha suppression of the LXR-sterol regulatory element binding protein-1c pathway, we propose the presence of an intricate network of nutritional transcription factors with mutual interactions, resulting in efficient reciprocal regulation of lipid degradation and lipogenesis. PMID- 12730334 TI - Effects of plasma adrenaline on hormone-sensitive lipase at rest and during moderate exercise in human skeletal muscle. AB - We investigated the effect of increased plasma adrenaline on hormone-sensitive lipase (HSL) activity and extracellular regulated kinase (ERK) 1/2 phosphorylation during exercise. Seven untrained men rested for 20 min and exercised for 10 min at 60 % peak pulmonary oxygen uptake on three occasions: with adrenaline infusion throughout rest and exercise (ADR), with no adrenaline infusion (CON) and with adrenaline infusion commencing after 3 min of exercise (EX+ADR). Muscle samples were obtained at rest before (Pre, -20 min) and after (0 min) infusion, and at 3 and 10 min of cycling. Exogenous adrenaline infusion increased (P < 0.05) plasma adrenaline at rest during ADR, which resulted in greater HSL activity (Pre, 2.14 +/- 0.10 mmol min-1 (kg dry matter (dm))-1; 0 min, 2.74 +/- 0.20 mmol min-1 (kg dm)-1). Subsequent exercise had no effect on HSL activity. During exercise in CON, HSL activity was increased (P < 0.05) above rest at 3 min but was not increased further by 10 min. The infusion of exogenous adrenaline at 3 min of exercise in EX+ADR resulted in a marked elevation in plasma adrenaline levels (3 min, 0.57 +/- 0.12 nM; 10 min, 10.08 +/- 0.84 nM) and increased HSL activity by 25 %. HSL activity at 10 min was greater (P < 0.05) in EX+ADR compared with CON. There were no changes between trials in the plasma concentrations of insulin and free fatty acids (FFA) and the muscle contents of free AMP, all putative regulators of HSL activity. ERK1/2 phosphorylation increased at 3 min in CON and EX+ADR. Because HSL activity did not increase during exercise when adrenaline was infused prior to exercise (ADR) and because HSL activity increased when adrenaline was infused during exercise (EX+ADR), we conclude that (1) high adrenaline levels can stimulate HSL activity regardless of the metabolic milieu and (2) large increases in adrenaline during exercise, independent of changes in other putative regulators, are able to further stimulate the contraction-induced increase in HSL activity. The results also demonstrate that increased ERK 1/2 phosphorylation coincides with elevated HSL activity, indicating that ERK 1/2 may mediate the contraction-induced increase in HSL activity early in exercise. PMID- 12730335 TI - Enhanced GABA(A) receptor-mediated activity following activation of NMDA receptors in Cajal-Retzius cells in the developing mouse neocortex. AB - Cajal-Retzius (CR) cells are among the earliest generated population of neurons in the developing neocortex and have been implicated in regulating cortical lamination. In rodents, CR cells are transient, being present only up to 2-3 weeks after birth. Although previous electrophysiological studies have demonstrated the presence of NMDA and GABAA receptors in CR cells, little is known about the functional properties of these receptors. Using whole-cell patch clamp techniques in neocortical slices, we confirmed the presence of D aminophosphonovaleric acid (APV)- and ifenprodil-sensitive NMDA receptors, and found that the functional expression of this receptor subtype is strain specific. The NMDA-induced response was consistently accompanied by overriding current transients that were blocked by APV and ifenprodil. In addition, bicuculline readily abolished these transients without affecting the NMDA-induced current response. The generation of these overriding current transients was dependent upon intracellular Ca2+ and was prevented by dialysis with the high-affinity Ca2+ chelator BAPTA. Overall, this study uncovered a synergistic interaction between these receptors, whereby activation of NMDA receptors leads to enhanced GABAA receptor-mediated activity through a Ca2+-dependent mechanism. PMID- 12730336 TI - Failure of systemic hypoxia to blunt alpha-adrenergic vasoconstriction in the human forearm. AB - Systemic hypoxia in humans evokes forearm vasodilatation despite significant reflex increases in sympathetic vasoconstrictor nerve activity and noradrenaline spillover. We sought to determine whether post-junctional alpha-adrenergic vasoconstrictor responsiveness to endogenous noradrenaline release is blunted during systemic hypoxia. To do so, we conducted a two-part study in healthy young adults. In protocol 1, we measured forearm blood flow (FBF; venous occlusion plethysmography) and calculated the vascular conductance (FVC) responses to brachial artery infusions of two doses of tyramine (evokes endogenous noradrenaline release) in 10 adults during normoxia and mild systemic hypoxia (85 % O2 saturation; pulse oximetry of the earlobe). Systemic hypoxia evoked significant forearm vasodilatation as indicated by the increases in FBF and FVC (approximately 20-23 %; P < 0.05). The low and high doses of tyramine evoked significant reductions in FVC (vasoconstriction) that were similar in magnitude during normoxia (-29 +/- 3 and -53 +/- 4 %) and mild hypoxia (-35 +/- 4 and -58 +/- 3 %; P = 0.33). In protocol 2, forearm vasoconstrictor responses to the high dose of tyramine were determined in eight young adults during normoxia and during graded levels of systemic hypoxia (85, 80 and 75 % O2 saturation). The reductions in FVC were similar during normoxia (-59 +/- 2 %) and the three levels of hypoxia (85 % O2 saturation, -64 +/- 3 %; 80 % O2 saturation, -62 +/- 1 %; 75 % O2 saturation, -61 +/- 3 %; P = 0.37). In both protocols, the tyramine-induced increases in deep venous noradrenaline concentrations were similar during normoxia and all levels of hypoxia. Our results demonstrate that post-junctional alpha-adrenergic receptor vasoconstrictor responsiveness to endogenous noradrenaline release is not blunted during mild-to-moderate systemic hypoxia in healthy humans. PMID- 12730337 TI - Reduced nephron number in adult sheep, hypertensive as a result of prenatal glucocorticoid treatment. AB - There is some evidence, mainly from rodent studies, that any factor which alters the final total number of nephrons formed, during nephrogenesis, will result in hypertension in adult life. Sheep, programmed to become hypertensive by exposure to synthetic glucocorticoid (dexamethasone, 0.48 mg h-1, for 48 h) early in development (~27 days of gestation), were killed at 7 years of age, and had nephron counting performed by unbiased stereology. Mean arterial pressure was 83 +/- 4 mmHg in the dexamethasone (DEX) group (n = 5), and 73 +/- 5 in the control (CON; n = 7; P < 0.05). The total nephron number, in the right kidney (249 070 +/ 14 331; n = 5) was significantly lower (P < 0.01) than that of controls (402 787 +/- 30 458; n = 7). Mean glomerular volume was larger in the DEX than the CON group (P < 0.01), but there was no significant difference in the sclerosis index between the two groups. Low nephron number was associated with grossly enlarged and dilated proximal tubules and greater accumulation of collagen type I and type III in the tubular interstitium and periadventitia of the renal cortical vessels. These data suggest that the hypertensive programming effect of glucocorticoid treatment, early in kidney development, results, at least in part, from impaired nephrogenesis. PMID- 12730338 TI - Phosphorylation-induced modulation of pNBC1 function: distinct roles for the amino- and carboxy-termini. AB - The human NBC1 (SLC4A4) gene encodes the electrogenic sodium bicarbonate cotransporters kNBC1 and pNBC1, which are highly expressed in the kidney and pancreas, respectively. The HCO3-:Na+ stoichiometry of these cotransporters is an important determinant of the direction of ion flux. Recently we showed in a mouse proximal tubule (mPCT) cell line expressing kNBC1, that 8-Br-cAMP shifts the stoichiometry of the cotransporter from 3:1 to 2:1 via protein kinase A (PKA) dependent phosphorylation of Ser982. pNBC1 has the identical carboxy-terminal consensus phosphorylation PKA site (KKGS1026), and an additional site in its amino-terminus (KRKT49). In this study we determined the potential role of these sites in regulating the function of pNBC1. The results demonstrated that in mPCT cells expressing pNBC1, PKA-dependent phosphorylation of Ser1026 following 8-Br cAMP treatment shifted the stoichiometry from 3:1 to 2:1. The effect was electrostatic in nature as replacing Ser1026 with Asp resulted in a similar stoichiometry shift. In addition to shifting the stoichiometry, 8-Br-cAMP caused a significant increase in the 4,4'-dinitrostilbene-2,2'-disulfonic acid (DNDS) sensitive basolateral membrane conductance (GDS) of cells expressing pNBC1, but not kNBC1. Although, the effect did not involve phosphorylation of Thr49, which was endogenously phosphorylated, replacing this residue with Asp or Ala abolished the 8-Br-cAMP-induced increase in GDS. In the mPEC pancreatic duct cell line, where endogenous pNBC1 functions with a HCO3-:Na+ stoichiometry of 2:1, 8-Br-cAMP increased GDS by ~90 % without altering the stoichiometry or inducing phosphorylation of the cotransporter. The results demonstrate that phosphorylation of Ser1026 mediates the cAMP-dependent shift in the stoichiometry of pNBC1, whereas Thr49 plays an essential role in the cAMP-induced increase in GDS. PMID- 12730339 TI - Stimulation of sodium pump restores membrane potential to neurons excited by glutamate in zebrafish distal retina. AB - Glutamate either depolarizes or hyperpolarizes retinal neurons. Those are the initial and primary effects. Using a voltage probe (oxonol, DiBaC4 (5)) to study dissociated zebrafish retinal neurons, we find a secondary, longer-term effect: a post-excitatory restoration of membrane potential, termed after-hyperpolarization (AHP). AHP occurs only in neurons that are depolarized by glutamate and typically peaks about 5 min after glutamate application. AHP is seen in dissociated horizontal cells (HCs) and hyperpolarizing, or OFF type, bipolar cells (HBCs). These cells commonly respond with only an AHP component. AHP never occurs in depolarizing, or ON type, bipolar cells (DBCs), which are cell types hyperpolarized by glutamate. AHP is blocked by 6-cyano-7-nitroquinoxaline-2,3 dione (CNQX). It is evoked by kainate, AMPA and the AMPA-selective agonist (S)-5 fluorowillardiine, but not by NMDA, D-aspartate, the kainate-selective agonist SYM 2081 or by DL-2-amino-4-phosphonobutyric acid (DL-AP4). Cells with exclusively AHP responses are tonically depolarized. Resting potentials can be restored by nifedipine, suggesting a tonic, depolarizing action of L-type Ca2+ channels. However AHP is not blocked by nifedipine and is insensitive to [Cl-]o. AHP is blocked by Li+o substitution for Na+o and by ouabain. A mechanism is proposed in which Na+ entering through ionotropic AMPA channels stimulates Na+,K+ ATPase, which, by electrogenic action, restores membrane potential, generating the AHP response. Patterns of ATPase immunoreactivity support localization in the outer plexiform layer (OPL) as cone pedicles, HCs and BCs were positively labelled. Labelling was weaker in the inner plexiform layer (IPL) than in nuclear layers, though two IPL bands of immunoreactive BC terminals could be discerned, one in sublamina a and the other in sublamina b. Persistent stimulation of distal retina by photoreceptor glutamate may induce increased expression and activity of Na+,K+-ATPase, with a consequent impact on distal glutamate responses. PMID- 12730340 TI - Neuropeptide Y and peptide YY inhibit excitatory synaptic transmission in the rat dorsal motor nucleus of the vagus. AB - Pancreatic polypeptides (PPs) such as neuropeptide Y (NPY) and peptide YY (PYY) exert profound, vagally mediated effects on gastrointestinal (GI) motility and secretion. Whole-cell patch clamp recordings were made from brainstem slices containing identified GI-projecting rat dorsal motor nucleus of the vagus (DMV) neurons to determine the mechanism of action of PPs. Electrical stimulation of nucleus tractus solitarii (NTS) induced excitatory postsynaptic currents (EPSCs) that were reduced in a concentration-dependent manner by NPY and PYY (both at 0.1 300 nM) in 65 % of the neurons. An increase in the paired-pulse ratio without changes in the postsynaptic membrane input resistance or EPSC rise and decay time suggested that the effects of PPs on EPSCs were due to actions at presynaptic receptors. The Y1 and Y2 receptor selective agonists [Leu31,Pro34]NPY and NPY(3 36) (both at 100 nM) mimicked the inhibition of NPY and PYY on the EPSC amplitude. The effects of 100 nM NPY, but not PYY, were antagonized partially by the Y1 receptor selective antagonist BIBP3226 (0.1 micro M). In addition, the inhibition of the EPSC amplitude induced by NPY, but not PYY, was attenuated partially by pretreatment with the alpha2 adrenoceptor antagonist yohimbine (10 micro M), and occluded partially by the alpha2 adrenoceptor agonist UK14,304 (10 micro M) as well as by pretreatment with reserpine. Pretreatment with a combination of BIBP3226 and yohimbine almost completely antagonized the NPY mediated effects on EPSCs. Contrary to the inhibition of EPSCs, perfusion with PPs had no effect on the amplitude of inhibitory postsynaptic currents (IPSCs) and a minimal effect on a minority of DMV neurons. Differences in the receptor subtypes utilized and in the mechanism of action of NPY and PYY may indicate functional differences in their roles within the circuitry of the dorsal vagal complex (DVC). PMID- 12730341 TI - Baroreflex-mediated changes in cardiac output and vascular conductance in response to alterations in carotid sinus pressure during exercise in humans. AB - We sought to quantify the contribution of cardiac output (Q) and total vascular conductance (TVC) to carotid baroreflex (CBR)-mediated changes in mean arterial pressure (MAP) during mild to heavy exercise. CBR function was determined in eight subjects (25 +/- 1 years) at rest and during three cycle exercise trials at heart rates (HRs) of 90, 120 and 150 beats min-1 performed in random order. Acute changes in carotid sinus transmural pressure were evoked using 5 s pulses of neck pressure (NP) and neck suction (NS) from +40 to -80 Torr (+5.33 to -10.67 kPa). Beat-to-beat changes in HR and MAP were recorded throughout. In addition, stroke volume (SV) was estimated using the Modelflow method, which incorporates a non linear, three-element model of the aortic input impedance to compute an aortic flow waveform from the arterial pressure wave. The application of NP and NS did not cause any significant changes in SV either at rest or during exercise. Thus, CBR-mediated alterations in Q were solely due to reflex changes in HR. In fact, a decrease in the carotid-HR response range from 26 +/- 7 beats min-1 at rest to 7 +/- 1 beats min-1 during heavy exercise (P = 0.001) reduced the contribution of Q to the CBR-mediated change in MAP. More importantly, at the time of the peak MAP response, the contribution of TVC to the CBR-mediated change in MAP was increased from 74 +/- 14 % at rest to 118 +/- 6 % (P = 0.017) during heavy exercise. Collectively, these findings indicate that alterations in vasomotion are the primary means by which the CBR regulates blood pressure during mild to heavy exercise. PMID- 12730342 TI - Thin filament activation and unloaded shortening velocity of rabbit skinned muscle fibres. AB - The unloaded shortening velocity of skinned rabbit psoas muscle fibres is sensitive to [Ca2+]. To determine whether Ca2+ affects the unloaded shortening velocity via regulation of crossbridge kinetics or crossbridge number, the shortening velocity was measured following changes in either [Ca2+] or the number of active thin filament regulatory units. The native troponin C (TnC) was extracted and replaced with either cardiac TnC (cTnC) or a mixture of cTnC and an inactive mutant cardiac TnC (CBMII TnC). The unloaded shortening velocity of the cTnC-replaced fibres was determined at various values of [Ca2+] and compared with different cTnC:CBMII TnC ratios at a saturating [Ca2+]. If Ca2+ regulates the unloaded shortening velocity via kinetic modulation, differences in the velocity tension relationship between the cTnC fibres and the cTnC:CBMII TnC fibres would be apparent. Alternatively, Ca2+ control of the number of active crossbridges would yield similar velocity-tension relationships when comparing the cTnC and cTnC:CBMII TnC fibres. The results show a decline in the unloaded shortening velocity that is determined by the relative tension, defined as the level of thin filament activation, rather than the [Ca2+]. Furthermore, at lower levels of relative tension, the reduction in unloaded shortening is not the result of changes in any cooperative effects of myosin on Ca2+ binding to the thin filament. Rather, it may be related to a decrease in crossbridge-induced activation of the thin filament at the level of the individual regulatory unit. In summary, the results suggest that Ca2+ regulates the unloaded shortening velocity in skinned fibres by reducing the number of crossbridges able to productively bind to the thin filament without affecting any inherent property of the myosin. PMID- 12730343 TI - Multifunctional cells of mouse anterior pituitary reveal a striking sexual dimorphism. AB - The existence of cells storing and secreting two different anterior pituitary (AP) hormones (polyhormonal cells) or responding to several hypothalamic releasing hormones (HRHs) (multiresponsive cells) has been reported previously. These multifunctional cells could be involved in paradoxical secretion (AP hormone secretion evoked by a non-corresponding HRH) and transdifferentiation (phenotypic switch between mature cell types without cell division). Despite their putative physiological relevance, a comprehensive characterization of multifunctional AP cells is lacking. Here we combine calcium imaging (to assess responses to the four HRHs) and multiple sequential immunoassay of the six AP hormones in the same individual cells to perform a complete phenotypic characterization of mouse AP cells. Polyhormonal and multiresponsive cells were identified within all five AP cell types. They were scarce in the more abundant cell types, somatotropes and lactotropes, but quite frequent in corticotropes and gonadotropes. Cells with mixed phenotypes were the rule rather than the exception in thyrotropes, where 56-83 % of the cells stored two to five different hormones. Multifunctional AP cells were much more abundant in females than in males, indicating that the hormonal changes associated with the sexual cycle may promote transdifferentiation. As the phenotypic analysis was performed here after stimulation with HRHs, the fraction of polyhormonal cells might have been underestimated. With this limitation, the polyhormonal cells detected here responded to the HRHs less than the monohormonal ones, suggesting that they might contribute less than expected a priori to paradoxical secretion. Overall, our results reveal a striking sexual dimorphism, the female pituitary being much more plastic than the male pituitary. PMID- 12730344 TI - Dynamic imaging of somatosensory cortical activity in the rat visualized by flavoprotein autofluorescence. AB - We used autofluorescence of mitochondrial flavoproteins to image cortical neural activity in the rat. Green autofluorescence in blue light was examined in slices obtained from rat cerebral cortex. About half of the basal autofluorescence was modulated by the presence or absence of O2 or glucose in the medium. Repetitive electrical stimulation at 20 Hz for 1 s produced a localized fluorescence increase in the slices. The amplitude of the increase was 27 +/- 2 % (mean +/- S.D., n = 35). Tetrodotoxin or diphenyleneiodonium, an inhibitor of flavoproteins, blocked the autofluorescence responses. The autofluorescence responses were not observed in slices perfused with calcium-, glucose- or O2-free medium. In the primary somatosensory cortex of rats anaesthetized with urethane (1.5 g kg-1, I.P.), an activity-dependent increase in autofluorescence of 20 +/- 4 % (n = 6) was observed after electrical cortical stimulation at 100 Hz for 1 s, and an increase of 2.6 +/- 0.5 % (n = 33) after vibratory skin stimulation at 50 Hz for 1 s applied to the plantar hindpaw. These responses were large enough to allow visualization of the neural activity without having to average a number of trials. The distribution of the fluorescence responses after electrical or vibratory skin stimulation was comparable to that of the cortical field potentials in the same rats. The fluorescence responses were followed by an increase in arterial blood flow. The former were resistant to an inhibitor of nitric oxide synthase, while the latter was inhibited. Thus, activity-dependent changes in the autofluorescence of flavoproteins are useful for functional brain imaging in vivo. PMID- 12730345 TI - ATP triggers intracellular Ca2+ release in type II cells of the rat carotid body. AB - Using a Ca2+-imaging technique, we studied the action of ATP on the intracellular Ca2+ concentration ([Ca2+]i) of fura-2-loaded mixtures of type I and type II cells dissociated from rat carotid bodies. ATP (100 micro M) triggered a transient rise in [Ca2+]i in the spindle-shaped type II (sustentacular) cells, but not the ovoid type I (glomus) cells. When challenged with ionomycin (1 micro M), no amperometry signal could be detected from the ATP-responsive type II cells, suggesting that these cells lacked catecholamine-containing granules. In contrast, KCl depolarization triggered robust quantal catecholamine release from type I cells that were not responsive to ATP. In type II cells voltage clamped at -70 mV, the ATP-induced [Ca2+]i rise was not accompanied by any current change, suggesting that P2X receptors are not involved. The ATP-induced Ca2+ signal could be observed in the presence of Ni2+ (a blocker of voltage-gated Ca2+ channels) or in the absence of extracellular Ca2+, indicating that Ca2+ release from intracellular stores was the dominant mechanism. The order of purinoreceptor agonist potency in triggering the [Ca2+]i rise was UTP > ATP > 2-methylthioATP >> alpha,beta-methyleneATP, implicating the involvement of P2Y2 receptors. In carotid body sections, immunofluorescence revealed localization of P2Y2 receptors on spindle-shaped type II cells that partially enveloped ovoid type I cells. Since ATP is released from type I cells during hypoxia, we suggest that the ATP induced Ca2+ signal in type II cells can mediate paracrine interactions within the carotid bodies. PMID- 12730346 TI - Development of synchronized activity of cranial motor neurons in the segmented embryonic mouse hindbrain. AB - Spontaneous electrical activity synchronized among groups of related neurons is a widespread and important feature of central nervous system development. Among the many places from which spontaneous rhythmic activity has been recorded early in development are the cranial motor nerve roots that exit the hindbrain, the motor neuron pool that, at birth, will control the rhythmic motor patterns of swallow, feeding and the oral components of respiratory behaviour. Understanding the mechanism and significance of this hindbrain activity, however, has been hampered by the difficulty of identifying and recording from individual hindbrain motor neurons in living tissue. We have used retrograde labelling to identify living cranial branchiomeric motor neurons in the hindbrain, and [Ca2+]i imaging of such labelled cells to measure spontaneous activity simultaneously in groups of motor neuron somata. We find that branchiomeric motor neurons of the trigeminal and facial nerves generate spontaneous [Ca2+]i transients throughout the developmental period E9.5 to E11.5. During this two-day period the activity changes from low-frequency, long-duration events that are tetrodotoxin insensitive and poorly coordinated among cells, to high-frequency short-duration events that are tetrodotoxin sensitive and tightly coordinated throughout the motor neuron population. This early synchronization may be crucial for correct neuron-target development. PMID- 12730347 TI - C-type inactivation involves a significant decrease in the intracellular aqueous pore volume of Kv1.4 K+ channels expressed in Xenopus oocytes. AB - Channels are water-filled membrane-spanning proteins, which undergo conformational changes as they gate, i.e. open or close. These conformational changes affect both the shape of the channel and the volume of the water-filled pore. We measured the changes in pore volume associated with activation, deactivation, C-type inactivation and recovery in an N-terminal-deleted mutant of the Kv1.4 K+ channel (Kv1.4DeltaN) expressed in Xenopus oocytes. We used giant patch and cut-open oocyte voltage clamp techniques and applied solutes which are too large to enter the pore mouth to exert osmotic pressure and thus favour smaller pore volume conformations. Applied intracellular osmotic pressure (300 mM sucrose) sped inactivation (time constants (tauinactivation): control, 0.66 +/- 0.09 s; hyperosmotic solution, 0.29 +/- 0.04 s; n = 5, P < 0.01), sped deactivation (taudeactivation: control, 18.8 +/- 0.94 ms; hyperosmotic solution, 8.01 +/- 1.92 ms; n = 5, P < 0.01), and slowed activation (tauactivation: control, 1.04 +/- 0.05 ms; hyperosmotic solution, 1.96 +/- 0.31 ms; n = 5, P < 0.01). These effects were reversible and solute independent. We estimated the pore volume change on inactivation to be about 4500 A3. Osmotic pressure had no effect when applied extracellularly. These data suggest that the intracellular side of the pore closes during C-type inactivation and the volume change is similar to that associated with activation or deactivation. This is also similar to the pore volume estimated from the crystal structure of KcsA and MthK K+ channels. Intracellular osmotic pressure also strongly inhibited re-opening currents associated with recovery from inactivation, which is consistent with a physical similarity between the C-type inactivated and resting closed state. PMID- 12730348 TI - Action potentials in basal and oblique dendrites of rat neocortical pyramidal neurons. AB - Basal and oblique dendrites comprise ~2/3 of the total excitable membrane in the mammalian cerebral cortex, yet they have never been probed with glass electrodes, and therefore their electrical properties and overall impact on synaptic processing are unknown. In the present study, fast multi-site voltage-sensitive dye imaging combined with somatic recording was used to provide a detailed description of the membrane potential transients in basal and oblique dendrites of pyramidal neurons during single and trains of action potentials (APs). The optical method allowed simultaneous measurements from several dendrites in the visual field up to 200 microm from the soma, thus providing a unique report on how an AP invades the entire dendritic tree. In contrast to apical dendrites, basal and oblique branches: (1) impose very little amplitude and time course modulation on backpropagating APs; (2) are strongly invaded by the somatic spike even when somatic firing rates reach 40 Hz (activity-independent backpropagation); and (3) do not exhibit signs of a 'calcium shoulder' on the falling phase of the AP. A compartmental model incorporating AP peak latencies and half-widths obtained from the apical, oblique and basal dendrites indicates that the specific intracellular resistance (Ri) is less than 100 omicron cm. The combined experimental and modelling results also provide evidence that all synaptic locations along basal and oblique dendrites, situated within 200 microm from the soma, experience strong and near-simultaneous (latency < 1 ms) voltage transients during somatic firing. The cell body, axon hillock and basal dendritic compartments achieve unique synchronization during each AP. Therefore, with respect to a retrograde signal (AP), basal and proximal oblique dendrites should be considered as an integral part of the axo-somatic compartment. PMID- 12730349 TI - Interleukin-2 increases activity of sarcoplasmic reticulum Ca2+-ATPase, but decreases its sensitivity to calcium in rat cardiomyocytes. AB - To further explore the role of interleukin-2 (IL-2) in cardiac function, we investigated its effects on the intracellular calcium transient and the activity of sarcoplasmic reticulum (SR) Ca2+-ATPase in rat cardiomyocytes. IL-2 (200 U/ml) decreased the amplitude of electrically stimulated and caffeine-induced intracellular Ca2+ transients in ventricular myocytes, but increased the end diastolic calcium level. IL-2 did not affect the sarcolemmal L-type Ca2+ channel activity. The activity of SR Ca2+-ATPase from IL-2-treated hearts increased in a dose-dependent manner, but the sarcolemmal Ca2+-ATPase activity did not change. After incubation of SR with ATP, the activity of SR Ca2+-ATPase from IL-2-treated hearts increased much more than that in the control group. The responsiveness of SR Ca2+-ATPase from IL-2-perfused hearts to the free calcium concentration was inhibited. The Ca2+ uptake and Ca2+ content were reduced in the SR vesicles prepared from IL-2-treated rat heart. Pretreatment with the kappa-opioid receptor antagonist nor-binaltorphimine (10 nM) attenuated the effect of IL-2 on the SR Ca2+-ATPase activity, SR Ca2+ uptake, and Ca2+ content. The activity of Ca2+ ATPase in SR isolated from untreated hearts did not change when IL-2 and SR were coincubated. Thus, we conclude that the decreased calcium transient induced by IL 2 results from reduced SR calcium release, which is due to decreased SR Ca2+ uptake mediated by cardiac kappa-opioid receptors, but not from reduced activity of the sarcolemmal L-type calcium channel. PMID- 12730350 TI - LY503430, a novel alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptor potentiator with functional, neuroprotective and neurotrophic effects in rodent models of Parkinson's disease. AB - Glutamate is the major excitatory transmitter in the brain. Recent developments in the molecular biology and pharmacology of the alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic acid (AMPA) subtype of glutamate receptors have led to the discovery of selective, potent, and systemically active AMPA receptor potentiators. These molecules enhance synaptic transmission and play important roles in plasticity and cognitive processes. In the present study, we first characterized a novel AMPA receptor potentiator, (R)-4'-[1-fluoro-1-methyl-2 (propane-2-sulfonylamino)-ethyl]-biphenyl-4-carboxylic acid methylamide (LY503430), on recombinant human GLUA1-4 and native preparations in vitro and then evaluated the potential neuroprotective effects of the molecule in rodent models of Parkinson's disease. Results indicated that submicromolar concentrations of LY503430 selectively enhanced glutamate-induced calcium influx into human embryonic kidney 293 cells transfected with human GLUA1, GLUA2, GLUA3, or GLUA4 AMPA receptors. The molecule also potentiated AMPA-mediated responses in native cortical, hippocampal, and substantia nigra neurons. We also report here that LY503430 provided dose-dependent functional and histological protection in animal models of Parkinson's disease. The neurotoxicity after unilateral infusion of 6-hydroxydopamine into either the substantia nigra or the striatum of rats and that after systemic 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine in mice were reduced. Interestingly, LY503430 also had neurotrophic actions on functional and histological outcomes when treatment was delayed until well after (6 or 14 days) the lesion was established. LY503430 also produced some increase in brain-derived neurotrophic factor in the substantia nigra and a dose-dependent increases in growth associated protein-43 (GAP-43) expression in the striatum. Therefore, we propose that AMPA receptor potentiators offer the potential of a new disease modifying therapy for Parkinson's disease. PMID- 12730351 TI - PAT5A: a partial agonist of peroxisome proliferator-activated receptor gamma is a potent antidiabetic thiazolidinedione yet weakly adipogenic. AB - PAT5A [5-[4-[N-(2-pyridyl)-(2S)-pyrrolidine-2 methoxyl]phenylmethylene[thiazolidine-2,4-dione, malic acid salt]], a chemically distinct unsaturated thiazolidinedione, activates peroxisome proliferator activated receptor gamma (PPARgamma) submaximally in vitro with the binding affinity approximately 10 times less than that of rosiglitazone, a highly potent thiazolidinedione. PAT5A reduces plasma glucose level and improves insulin sensitivity in insulin resistant db/db mice, similar to that of rosiglitazone, while exerting a relatively weak adipogenic effect. In contrast to rosiglitazone, PAT5A inhibits cholesterol and fatty acid biosynthesis suggesting that PAT5A possesses a unique receptor-independent non-PPAR related property. PAT5A induces qualitatively similar but quantitatively different protease digestion patterns and interacts with PPARgamma differently than rosiglitazone. PAT5A shows differential cofactor recruitment and gene activation than that of rosiglitazone. Thus, the partial agonism of PAT5A to PPARgamma together with its receptor independent effects may contribute to its antidiabetic potency similar to rosiglitazone in vivo despite reduced affinity for PPARgamma. These biological effects suggest that PAT5A is a PPARgamma modulator that activates some (insulin sensitization), but not all (adipogenesis), PPARgamma-signaling pathways. PMID- 12730352 TI - Continuous administration of the 5-hydroxytryptamine1A agonist (3-Chloro-4-fluoro phenyl)-[4-fluoro-4-[[(5-methyl-pyridin-2-ylmethyl) -amino]-methyl]piperidin-1 yl]-methadone (F 13640) attenuates allodynia-like behavior in a rat model of trigeminal neuropathic pain. AB - (3-Chloro-4-fluoro-phenyl)-[4-fluoro-4-[[(5-methyl-pyridin-2-ylmethyl)-amino] methyl]piperidin-1-yl]-methadone (F 13640) is a recently discovered high-efficacy 5-hydroxytryptamine (HT)1A receptor agonist that produces central analgesia through the neuroadaptive mechanisms of inverse tolerance and cooperation. In a rat model of trigeminal neuropathic pain, the chronic constriction injury of the infraorbital nerve causes allodynia-like behavior that develops within 2 weeks and remains stable thereafter. We report that early after surgery, during which time allodynia develops, the continuous 2-week infusion of 0.63 mg/day F 13640 inhibited the allodynia-like behavior, whereas 5 mg/day morphine showed no significant effect. When F 13640 infusion was initiated late after surgery, when allodynia was well established, it produced an antiallodynic effect that was apparent during the entire infusion period. In contrast, morphine infusion caused an initially marked antiallodynic effect to which tolerance developed within the 2-week infusion period. The GABA-B receptor agonist baclofen (1.06 mg/day) that has a recognized usefulness in the treatment of trigeminal neuralgia, demonstrated effectiveness in both conditions. The data are consistent with a theory of nociceptive signal transduction, as well as with previous data, in demonstrating the neuroadaptive mechanisms of inverse tolerance and cooperation. That is, in contrast with morphine, the antiallodynic effect induced by 5-HT1A receptor activation does not decay, but, if anything, grows with chronicity. Also, 5-HT1A receptor activation seemed to cooperate with nociceptive stimulation in, paradoxically, inducing an antiallodynic effect. The data presented here suggest that F 13640 may perhaps offer a lasting treatment of trigeminal neuralgia. PMID- 12730353 TI - A mutation in transmembrane domain II of the 5-hydroxytryptamine(3A) receptor stabilizes channel opening and alters alcohol modulatory actions. AB - Mutant 5-hydroxytryptamine (5-HT)3A receptors, in which changes were made at Ile294, position 16', of the second transmembrane domain, were assessed for alterations in macroscopic response kinetics and modulation by alcohols. Function of heterologously expressed receptors was measured in Xenopus oocytes in the two electrode voltage clamp configuration and in human embryonic kidney 293 cells using whole cell patch-clamp electrophysiological recordings with rapid drug application. Compared with the wild-type receptor, a decrease in the 5-HT EC50 value in the Ile294Thr mutant was observed, whereas an increase in the 5-HT EC50 value in the Ile294Leu mutant was measured. Ile294Thr receptors showed a marked reduction in the extent of desensitization. Ethanol and 2,2,2-trichloroethanol (TCEt) enhanced 5-HT-mediated currents in wild-type and Ile294Leu receptors, but inhibited or had little stimulatory effect in the Ile294Thr mutant. Kinetic analysis revealed that in the presence of TCEt, the slope of activation was unchanged in the Ile294Thr mutant and increased in the wild-type receptor. Alcohol cutoff was altered with wild-type = heptanol and Ile294Leu = hexanol. Kinetic changes in the Ile294Thr mutant that favor the open channel state, as well as reduction in the rate of channel activation in the presence of TCEt, likely underlie this mutant's altered response to n-chain alcohols. PMID- 12730354 TI - Suppressed prolactin response to dynorphin A1-13 in methadone-maintained versus control subjects. AB - Dynorphin A1-13, a shortened sequence of the natural peptide dynorphin A1-17, is a primarily kappa-opioid receptor-preferring peptide. Previously, we showed that dynorphin A1-13 administered to normal volunteers causes a prompt dose-dependent elevation in serum prolactin that may reflect a reduction in tuberoinfundibular dopaminergic tone. This study was conducted to determine whether tuberoinfundibular dopaminergic tone is reduced in methadone-maintained patients. Eight former heroin addicts on stable-dose methadone maintenance with no ongoing drug or alcohol abuse or dependence and 15 normal volunteer controls with no history of drug or alcohol dependence received dynorphin A1-13 intravenously at doses of 120 microg/kg and 500 microg/kg. Studies began one hour before methadone dosing to avoid the expected increase in prolactin that coincides with peak plasma levels of methadone. After intravenous dynorphin A1-13, a dose-response increase in serum prolactin, which peaked within 20 min, was observed in both groups. There was no difference in prolactin between the two groups at baseline or following a placebo. The prolactin response to each dose of dynorphin A1-13 was significantly lower in the methadone-maintained volunteers compared with the controls. These results suggest that tuberoinfundibular dopaminergic tone is altered in methadone-maintained subjects. It is unknown whether altered dopaminergic tone existed before opiate addiction, is a result of heroin addiction, or is reflective of methadone maintenance. Whether methadone maintained subjects also have decreased dopaminergic response to dynorphin and other kappa-opioid receptor ligands in mesolimbic-mesocortical and nigrostriatal dopaminergic systems cannot be determined from this study. PMID- 12730355 TI - Sigma-1 receptors (sigma(1) binding sites) form raft-like microdomains and target lipid droplets on the endoplasmic reticulum: roles in endoplasmic reticulum lipid compartmentalization and export. AB - The brain sigma-1 receptors can bind neurosteroids and psychotropic drugs, including neuroleptics and cocaine and are implicated in schizophrenia, depression, and drug dependence. In this study, we found that sigma-1 receptors specifically target lipid storage sites (lipid droplets) on the endoplasmic reticulum by forming a distinct class of lipid microdomains. Both endogenously expressing sigma-1 receptors and transfected C-terminally enhanced yellow fluorescent protein (EYFP)-tagged sigma-1 receptors (Sig-1R-EYFP) target unique "ring-like" structures associated with endoplasmic reticulum reticular networks in NG108-15 cells. The ring-like structures contain neutral lipids and are enlarged by the oleate treatment, indicating that they are endoplasmic reticulum associated lipid droplets (ER-LDs). sigma-1 receptors colocalize with caveolin-2, a cholesterol-binding protein in lipid rafts on the ER-LDs, but not with adipocyte differentiation-related protein (ADRP), a cytosolic lipid droplet (c LD)-specific protein. When the double-arginine ER retention signal on the N terminus of sigma-1 receptors is truncated, sigma-1 receptors no longer exist on ER-LDs, but predominantly target c-LDs, which contain ADRP. sigma-1 receptors on ER-LDs form detergent-resistant raft-like lipid microdomains, the buoyancy of which is different from that of plasma membrane lipid rafts. (+)-Pentazocine causes sigma-1 receptors to disappear from the microdomains. N-Terminally EYFP tagged sigma-1 receptors (EYFP-Sig-1R) failed to target ER-LDs. EYFP-Sig-1R transfected cells showed an unrestricted distribution of neutral lipids all over the endoplasmic reticulum network, decreases in c-LDs and cholesterol in plasma membranes, and the bulbous aggregation of endoplasmic reticulum. Thus, sigma-1 receptors are unique endoplasmic reticulum proteins that regulate the compartmentalization of lipids on the endoplasmic reticulum and their export from the endoplasmic reticulum to plasma membrane and c-LDs. PMID- 12730356 TI - Intracellular dynamics of sigma-1 receptors (sigma(1) binding sites) in NG108-15 cells. AB - The sigma-1 receptors bind diverse kinds of psychoactive compounds, including cocaine, and translocate upon stimulation by these compounds. However, the exact intracellular localization and dynamics of sigma-1 receptors have been unclear. We recently found that sigma-1 receptors specifically localize on cholesterol enriched loci on the endoplasmic reticulum (ER) membrane that function as neutral lipid storage sites (i.e., ER lipid droplets or ER-LDs) from which neutral lipids bud out to form cytosolic lipid droplets. By combining immunocytochemistry and real-time monitoring of enhanced yellow fluorescent protein (EYFP)-tagged sigma-1 receptors (Sig-1R-EYFP) in living cells, we characterized the sigma-1 receptor translocation in this study. (+)-Pentazocine, a selective sigma-1 receptor agonist, causes a significant decrease of sigma-1 receptors in ER-LDs and a diffused distribution of sigma-1 receptors over the entire endoplasmic reticulum reticular network in NG108-15 cells. In the presence of sigma-1 receptor agonists, Sig-1R-EYFP move out from ER-LDs and slide along the endoplasmic reticulum network toward nuclear envelope and the tip of neurites. Fluorescence recovery after photobleaching analysis demonstrates that Sig-1R-EYFP on endoplasmic reticulum reticular network are highly mobile compared with those in ER-LDs. A sucrose gradient fractionation study shows that (+)-pentazocine shifts sigma-1 receptors from ER-LD membranes to higher density membranes. These results indicate that sigma-1 receptors localize on ER-LDs and upon stimulation translocate on continuous endoplasmic reticulum reticular network toward peripheries of cells. Because sigma-1 receptors specifically target ER lipid storage sites and compartmentalize neutral lipids therein, these results suggest that sigma-1 receptors' dynamic translocation might affect lipid transport and distribution in neuronal cells. PMID- 12730357 TI - Neuroprotective effects of (S)-N-[4-[4-[(3,4-Dihydro-6-hydroxy-2,5,7,8 tetramethyl-2H-1-benzopyran-2-yl)carbonyl]-1-piperazinyl]phenyl]-2 thiophenecarboximid-amide (BN 80933), an inhibitor of neuronal nitric-oxide synthase and an antioxidant, in model of transient focal cerebral ischemia in mice. AB - Nitric oxide (NO) and reactive oxygen species are both implicated in neuronal death due to cerebral ischemia. BN 80933, an original compound associating an inhibitor of neuronal NO synthase with an antioxidant, has been shown to reduce functional and histological damage in rat submitted to cerebral ischemia. The aim of the present study was to confirm these results in mice and to further examine the effects of BN 80933 on inflammatory response, including blood-brain barrier (BBB) disruption, brain edema, and neutrophil infiltration after transient middle cerebral artery occlusion (MCAO). Intravenous administration of BN 80933 at 3 and 10 mg/kg 3 h after MCAO significantly reduced by 26 to 36% the infarct volume evaluated 24 and 48 h after ischemia, and improved the neurological score. Furthermore, BN 80933 at both dosages decreased by 42 to 75% the extravasation of Evans blue in brain parenchyma observed 24 h after ischemia. This reduction in BBB disruption was associated with decreased brain edema as demonstrated by the 37% reduction in brain water content induced by BN 80933 at 3 mg/kg 24 h after MCAO. Neutrophil infiltration in brain parenchyma, evaluated by the myeloperoxidase activity, was also reduced by 45 to 56% in animals treated with BN 80933 at 3 and 10 mg/kg. Together, these results extend the protective capacity of BN 80933 against brain ischemic injury and confirm that BN 80933 represents a promising treatment for stroke. PMID- 12730358 TI - Nifedipine-mediated mobilization of intracellular calcium stores increases spontaneous neurotransmitter release at neonatal rat motor nerve terminals. AB - The modulation of spontaneous release of acetylcholine by specific Ca2+ channel blockers was studied at neonatal rat neuromuscular junction. During early postnatal periods (0-4 days), blockers of N- and P/Q-type Ca2+ channels did not affect miniature endplate potential (MEPP) frequency. Unexpectedly, treatment with the L-type Ca2+ channel antagonist nifedipine, although not when treated with isradipine, nitrendipine, or calciseptine, resulted in strong increase in MEPP frequency. The potentiation effect of nifedipine was dose-dependent with a 56-fold maximum effect with 15 microM. The effect decreased during the first two postnatal weeks and disappeared by the third. The effect of nifedipine was not dependent on extracellular Ca2+ and was not altered by the presence of other Ca2+ channel blockers. In contrast, it was abolished by depleting intracellular Ca2+ stores with 2 microM thapsigargin and was partially inhibited by 10 microM ryanodine. In conclusion, we report a new ryanodine receptor-mediated effect of nifedipine on neonatal neuromuscular junction that may indicate the developmental expression of a specific receptor channel that interacts with intracellular Ca2+ stores. This effect of nifedipine should also be considered when using this drug as either a therapeutic or a research tool. PMID- 12730359 TI - Effects of in vivo lipopolysaccharide infusion on vasoconstrictor function of rat isolated mesentery, kidney, and aorta. AB - Continuous infusion of lipopolysaccharide (LPS) into conscious rats elicits regionally selective cardiovascular disturbances. The aim of the present study was to assess contractile function in different vascular preparations (renal, mesenteric, and thoracic aorta) taken from rats infused with LPS for 2 or 24 h. Sustained responses to continuous infusion of methoxamine but not to KCl were reduced in the aorta (at 2 and 24 h LPS) and mesentery (at 24 h LPS) but not in the renal vascular bed. In contrast, transient responses to bolus doses of methoxamine were unchanged in the mesentery. In Ca2+-imaging experiments with fura-2, challenge with a single concentration of methoxamine (10 microM, which showed an impaired contractile response at 24 h LPS) induced a rise in intracellular Ca2+ in the mesenteric artery that was not different from the control. Furthermore, in the aorta, the contractile response to caffeine was attenuated only in the 2 h LPS group. These results show that there is regional heterogeneity in in vitro vascular responsiveness in preparations taken from LPS infused rats. Thus, in mesenteric beds and aortae, but not renal beds, there is hypocontractility to methoxamine that is not due to a generalized inability of the smooth muscle to contract, which is evident with sustained but not transient application of agonist (mesentery) and which, in late endotoxemia (24 h LPS), does not appear to involve abnormalities in Ca2+ mobilization or entry. PMID- 12730360 TI - Evidence for a ceiling of cardioprotection with a nitric oxide donor-induced delayed preconditioning in rabbits. AB - Although extensive attention has been devoted to the window of preconditioning, only few studies investigated the efficacy of preconditioning against ischemia with increasing durations. To date, a "ceiling of protection" has been demonstrated to occur with early preconditioning but nothing is known about delayed preconditioning. Accordingly, the efficacy of a nitric oxide (NO)-donor induced delayed preconditioning was tested against ischemic insults of increasing duration. Accordingly, 65 rabbits received a 75-min intravenous infusion of either saline (control group), or an NO-donor (S-nitroso-N-acetylpenicillamine) at 3 microg/kg/min (SNAP-3 group) or 30 microg/kg/min (SNAP-30 group). Twenty four hours later, rabbits randomly underwent either a 15-, 20-, or a 30-min coronary artery occlusion (CAO). Infarct size was assessed after 72-h reperfusion (triphenyltetrazolium chloride staining, percentage of the area at risk). After 15-min CAO, both SNAP-3 and SNAP-30 reduced infarct size compared with control (10 +/- 3, 5 +/- 1 versus 29 +/- 8%, respectively; p < 0.05). After 20-min CAO, significant cardioprotection was only observed with SNAP-30 (29 +/- 4, 21 +/- 6 versus 36 +/- 2% for SNAP-3, SNAP-30 versus control, respectively). After 30-min CAO, both SNAP-3 and SNAP-30 failed to reduce infarct size (48 +/- 2, 50 +/- 5 versus 50 +/- 4% for SNAP-3, SNAP-30 versus control, respectively). In conclusion, this study demonstrates a dose-related ceiling of protection with delayed preconditioning induced by an NO donor. It supports that delayed preconditioning might exert its maximal beneficial effect with early reperfusion and this finding supports the necessary use of different durations of ischemia when investigating cardioprotective strategies. PMID- 12730361 TI - Oxidative stress-induced homologous recombination as a novel mechanism for phenytoin-initiated toxicity. AB - Although the mechanism(s) of phenytoin-initiated toxicity is unknown, phenytoin can be enzymatically bioactivated to a reactive intermediate leading to increased formation of reactive oxygen species, which can damage essential macromolecules, including DNA. The oxidation of DNA can induce DNA double-strand breaks (DSBs), which may be repaired through homologous recombination. Increased levels of DSBs may induce hyper-recombination, leading to deleterious genetic changes. We hypothesize that these genetic changes mediate phenytoin-initiated toxicity. To investigate this hypothesis we used a Chinese hamster ovary cell line containing a neo direct repeat recombination substrate to determine whether phenytoin initiated DNA oxidation increases homologous recombination. Cells were treated with 0 to 800 microM phenytoin for 5 or 24 h, and homologous recombination frequencies and recombinant product structures were determined. Phenytoin initiated DNA oxidation was determined by measuring the formation of 8-hydroxy-2' deoxyguanosine. We demonstrate that phenytoin increases both DNA oxidation and homologous recombination in a concentration- and time-dependent manner. All recombination products analyzed arose via gene conversion without associated crossover. Our data demonstrate that phenytoin-initiated DNA damage can induce homologous recombination, which may be a novel mechanism mediating phenytoin initiated toxicity. PMID- 12730363 TI - The nuclear pore complex protein ALADIN is mislocalized in triple A syndrome. AB - Triple A syndrome is a human autosomal recessive disorder characterized by an unusual array of tissue-specific defects. Triple A syndrome arises from mutations in a WD-repeat protein of unknown function called ALADIN (also termed Adracalin or AAAS). We showed previously that ALADIN localizes to nuclear pore complexes (NPCs), large multiprotein assemblies that are the sole sites of nucleocytoplasmic transport. Here, we present evidence indicating that NPC targeting is essential for the function of ALADIN. Characterization of mutant ALADIN proteins from triple A patients revealed a striking effect of these mutations on NPC targeting. A variety of disease-associated missense, nonsense, and frameshift mutations failed to localize to NPCs and were found predominantly in the cytoplasm. Microscopic analysis of cells from a triple A patient revealed no morphological abnormalities of the nuclei, nuclear envelopes, or NPCs. Importantly, these findings indicate that defects in NPC function, rather than structure, give rise to triple A syndrome. We propose that ALADIN plays a cell type-specific role in regulating nucleocytoplasmic transport and that this function is essential for the proper maintenance andor development of certain tissues. Our findings provide a foundation for understanding the molecular basis of triple A syndrome and may lead to unique insights into the role of nucleocytoplasmic transport in adrenal function and neurodevelopment. PMID- 12730362 TI - Identification and characterization of a G protein-coupled receptor for the neuropeptide proctolin in Drosophilamelanogaster. AB - Proctolin is a bioactive neuropeptide that modulates interneuronal and neuromuscular synaptic transmission in a wide variety of arthropods. We present several lines of evidence to propose that the orphan G protein-coupled receptor CG6986 of Drosophila is a proctolin receptor. When expressed in mammalian cells, CG6986 confers second messenger activation after proctolin application, with an EC(50) of 0.3 nM. In competition-based studies, the CG6986 receptor binds proctolin with high affinity (IC(50) = 4 nM). By microarray analysis, CG6986 transcript is consistently detected in head mRNA of different genotypes, and under different environmental conditions. By blot analysis, anti-CG6986 antibodies detect a band in tissue homogenates similar to the predicted size of the protein. Proctolin receptor immunosignals are found in the hindgut, heart, and in distinct neuronal populations of the CNS; such patterns correlate with previous demonstrations of proctolin biological activity, and in several instances, with areas of proctolin peptide immunosignals. The identification of a bona fide proctolin receptor provides the basis for a mechanistic analysis of this critical synaptic modulator. PMID- 12730364 TI - Protein kinase involved in lung injury susceptibility: evidence from enzyme isoform genetic knockout and in vivo inhibitor treatment. AB - Acute lung injury (ALI) associated with sepsis and iatrogenic ventilator-induced lung injury resulting from mechanical ventilation are major medical problems with an unmet need for small molecule therapeutics. Prevailing hypotheses identify endothelial cell (EC) layer dysfunction as a cardinal event in the pathophysiology, with intracellular protein kinases as critical mediators of normal physiology and possible targets for drug discovery. The 210,000 molecular weight myosin light chain kinase (MLCK210, also called EC MLCK because of its abundance in EC) is hypothesized to be important for EC barrier function and might be a potential therapeutic target. To test these hypotheses directly, we made a selective MLCK210 knockout mouse that retains production of MLCK108 (also called smooth-muscle MLCK) from the same gene. The MLCK210 knockout mice are less susceptible to ALI induced by i.p. injection of the endotoxin lipopolysaccharide and show enhanced survival during subsequent mechanical ventilation. Using a complementary chemical biology approach, we developed a new class of small molecule MLCK inhibitor based on the pharmacologically privileged aminopyridazine and found that a single i.p. injection of the inhibitor protected WT mice against ALI and death from mechanical ventilation complications. These convergent results from two independent approaches demonstrate a pivotal in vivo role for MLCK in susceptibility to lung injury and validate MLCK as a potential drug discovery target for lung injury. PMID- 12730365 TI - Assay of locus-specific genetic load implicates rare Toll-like receptor 4 mutations in meningococcal susceptibility. AB - As the central component of the human endotoxin sensor, Toll-like receptor 4 (TLR4) functions in the early detection and response to Gram-negative infection. We therefore examined a large collection of patients with meningococcal sepsis, comparing the frequency of rare TLR4 coding changes to those in an ethnically matched control population. TLR2 sequences were also acquired and compared. Total nucleotide variation at TLR4 and TLR2 loci was assayed by using a novel computational method. A total of 3.01 megabases of coding sequence was captured at these loci from white subjects with or without meningococcal disease. Authentic mutations were found and high-quality, bidirectional coverage was measured across the coding region by using mutationseeker, a program specifically designed to assay locus-specific genetic load. Using a method that obviates the confounding effect of linkage disequilibrium, we observed that rare heterozygous missense mutations of TLR4 contribute to the development of systemic meningococcal disease among white populations of the southern United Kingdom (P = 0.02; odds ratio 8.2). When results from all white populations were pooled, an overwhelmingly significant excess of such mutations was observed among individuals with disease (P = 2 x 10(-6); odds ratio 27.0). The common white TLR4 variant (TLR4B), synonymous TLR4 substitutions, and variant TLR2 alleles were not significantly over-represented among patients with systemic meningococcal infections. No single variant of TLR4 was significantly over-represented in the meningococcal population. Collectively, however, rare TLR4 coding variants were markedly over-represented. Sensing via TLR4 probably contributes to the early containment of meningococcal infection, and sensing defects create increased risk of disease. PMID- 12730366 TI - Public versus personal serotypes of a viral quasispecies. AB - Noncytopathic RNA viruses persist in their natural hosts at various levels as highly mutating quasispecies. They exhibit only one known serotype. In most inbred DBA2 mice infected with 2 x 10(4) or 2 x 10(6) plaque-forming units (pfu) of lymphocytic choriomeningitis virus (LCMV), the virus is transiently controlled below detectable levels measured with conventional assays (<1.7 pfu), but reemerges despite a common neutralizing Ab (nAb) response. Wild-type virus and cloned mutant viruses that had escaped polyclonal nAb responses in vivo induced nAb titers in new hosts that were usually cross-reactive; some sera were highly specific for certain mutants. The few mice that controlled LCMV infection for >170 days produced not only nAb against wild-type but also variably against many other mutants isolated from other mice with reemerging viremia. When DBA2 mice were immunized and boosted with 200 pfu of a LCMV mutant, the neutralizing Ab response was limited to the immunizing "personal" clone. Thus, in contrast to classical serotype-defined cytopathic viruses (e.g., polio viruses) that induce strictly non-cross-reactive nAb titers, LCMV, a noncytopathic RNA virus, represents a dynamic multiplicity of personal serological submutants. Together, these mutants form a generally recognized "public" serotype. These findings may help to explain aspects of human infections and Ab responses against hepatitis B virus, hepatitis C virus, and HIV. PMID- 12730367 TI - Tuning activation of the AMPA-sensitive GluR2 ion channel by genetic adjustment of agonist-induced conformational changes. AB - The (S)-2-amino-3-(3-hydroxy-5-methyl-4-isoxazole) propionic acid (AMPA) receptor discriminates between agonists in terms of binding and channel gating; AMPA is a high-affinity full agonist, whereas kainate is a low-affinity partial agonist. Although there is extensive literature on the functional characterization of partial agonist activity in ion channels, structure-based mechanisms are scarce. Here we investigate the role of Leu-650, a binding cleft residue conserved among AMPA receptors, in maintaining agonist specificity and regulating agonist binding and channel gating by using physiological, x-ray crystallographic, and biochemical techniques. Changing Leu-650 to Thr yields a receptor that responds more potently and efficaciously to kainate and less potently and efficaciously to AMPA relative to the WT receptor. Crystal structures of the Leu-650 to Thr mutant reveal an increase in domain closure in the kainate-bound state and a partially closed and a fully closed conformation in the AMPA-bound form. Our results indicate that agonists can induce a range of conformations in the GluR2 ligand binding core and that domain closure is directly correlated to channel activation. The partially closed, AMPA-bound conformation of the L650T mutant likely captures the structure of an agonist-bound, inactive state of the receptor. Together with previously solved structures, we have determined a mechanism of agonist binding and subsequent conformational rearrangements. PMID- 12730368 TI - Genomewide view of gene silencing by small interfering RNAs. AB - RNA interference (RNAi) is an evolutionarily conserved mechanism in plant and animal cells that directs the degradation of messenger RNAs homologous to short double-stranded RNAs termed small interfering RNA (siRNA). The ability of siRNA to direct gene silencing in mammalian cells has raised the possibility that siRNA might be used to investigate gene function in a high throughput fashion or to modulate gene expression in human diseases. The specificity of siRNA-mediated silencing, a critical consideration in these applications, has not been addressed on a genomewide scale. Here we show that siRNA-induced gene silencing of transient or stably expressed mRNA is highly gene-specific and does not produce secondary effects detectable by genomewide expression profiling. A test for transitive RNAi, extension of the RNAi effect to sequences 5' of the target region that has been observed in Caenorhabditis elegans, was unable to detect this phenomenon in human cells. PMID- 12730369 TI - Deletion of neuropeptide Y (NPY) 2 receptor in mice results in blockage of NPY induced angiogenesis and delayed wound healing. AB - Neuropeptide Y (NPY), a 36-aa peptide, is widely distributed in the brain and peripheral tissues. Whereas physiological roles of NPY as a hormoneneurotransmitter have been well studied, little is known about its other peripheral functions. Here, we report that NPY acts as a potent angiogenic factor in vivo using the mouse corneal micropocket and the chick chorioallantoic membrane (CAM) assays. Unlike vascular endothelial growth factor (VEGF), microvessels induced by NPY had distinct vascular tree-like structures showing vasodilation. This angiogenic pattern was similar to that induced by fibroblast growth factor-2, and the angiogenic response was dose-dependent. In the developing chick embryo, NPY stimulated vascular sprouting from preexisting blood vessels. When [Leu(31)Pro(34)]NPY, a NPY-based analogue lacking high affinity for the NPY Y(2) receptor but capable of stimulating both Y(1) and Y(5) receptors, was used in the corneal model, no angiogenic response could be detected. In addition, NPY failed to induce angiogenesis in Y(2) receptor-null mice, suggesting that this NPY receptor subtype was mediating the angiogenic signal. In support of this finding, the Y(2) receptor, but not Y(1), Y(4), or Y(5) receptors, was found to be widely expressed in newly formed blood vessels. Further, a delay of skin wound healing with reduced neovascularization was found in Y(2) receptor-null mice. These data demonstrate that NPY may play an important role in the regulation of angiogenesis and angiogenesis-dependent tissue repair. PMID- 12730371 TI - A novel class of gene controlling virulence in plant pathogenic ascomycete fungi. AB - Insertional mutants of the fungal maize pathogen Cochliobolus heterostrophus were screened for altered virulence. One mutant had 60% reduction in lesion size relative to WT but no other detectable change in phenotype. Analysis of sequence at the insertion site revealed a gene (CPS1) encoding a protein with two AMP binding domains. CPS1 orthologs were detected in all Cochliobolus spp. examined, in several other classes of ascomycete fungi, and in animals but not in basidiomycete fungi, bacteria, or plants. Phylogenetic analysis suggested that CPS1 represents a previously undescribed subset of adenylate-forming enzymes that have diverged from certain acyl-CoA ligases, which in bacteria are involved in biosynthesis of nonribosomal peptides or polyketidepeptide hybrids. Disruption of CPS1 caused reduced virulence of both race T and race O of C. heterostrophus on maize, of Cochliobolus victoriae on oats, and of Gibberella zeae on wheat. These results suggest that CPS1 functions as a general fungal virulence factor in plant pathogenic ascomycetes. PMID- 12730370 TI - An estrogen-dependent four-gene micronet regulating social recognition: a study with oxytocin and estrogen receptor-alpha and -beta knockout mice. AB - Estrogens control many physiological and behavioral processes, some of which are connected to reproduction. These include sexual and other social behaviors. Here we implicate four gene products in a micronet required for mammalian social recognition, through which an individual learns to recognize other individuals. Female mice whose genes for the neuropeptide oxytocin (OT) or the estrogen receptor (ER)-beta or ER-alpha had been selectively "knocked out" were deficient specifically in social recognition and social anxiety. There was a remarkable parallelism among results from three separate gene knockouts. The data strongly suggest the involvement in social recognition of the four genes coding for ER alpha, ER-beta, OT, and the OT receptor. We thus propose here a four-gene micronet, which links hypothalamic and limbic forebrain neurons in the estrogen control over the OT regulation of social recognition. In our model, estrogens act on the OT system at two levels: through ER-beta, they regulate the production of OT in the hypothalamic paraventricular nucleus, and through ER-alpha, they drive the transcription of the OT receptor in the amygdala. The proper operation of a social recognition mechanism allows for the expression of appropriate social behaviors, aggressive or affiliative. PMID- 12730373 TI - Digital genotyping and haplotyping with polymerase colonies. AB - Polymerase colony (polony) technology amplifies multiple individual DNA molecules within a thin acrylamide gel attached to a microscope slide. Each DNA molecule included in the reaction produces an immobilized colony of double-stranded DNA. We genotype these polonies by performing single base extensions with dye-labeled nucleotides, and we demonstrate the accurate quantitation of two allelic variants. We also show that polony technology can determine the phase, or haplotype, of two single- nucleotide polymorphisms (SNPs) by coamplifying distally located targets on a single chromosomal fragment. We correctly determine the genotype and phase of three different pairs of SNPs. In one case, the distance between the two SNPs is 45 kb, the largest distance achieved to date without separating the chromosomes by cloning or somatic cell fusion. The results indicate that polony genotyping and haplotyping may play an important role in understanding the structure of genetic variation. PMID- 12730372 TI - The two-component PhoR-PhoP system controls both primary metabolism and secondary metabolite biosynthesis in Streptomyces lividans. AB - The biosynthesis of most secondary metabolites in different bacteria is strongly depressed by inorganic phosphate. The two-component phoR-phoP system of Streptomyces lividans has been cloned and characterized. PhoR showed all of the characteristics of the membrane-bound sensor proteins, whereas PhoP is a member of the DNA-binding OmpR family. Deletion mutants lacking phoP or phoR-phoP, were unable to grow in minimal medium at low phosphate concentration (10 microM). Growth was fully restored by complementation with the phoR-phoP genes. Both S. lividans DeltaphoP and DeltaphoR-phoP deletion mutants were unable to synthesize extracellular alkaline phosphatase (AP) as shown by immunodetection with anti-AP antibodies and by enzymatic analysis, suggesting that the PhoR-PhoP system is required for expression of the AP gene (phoA). Synthesis of AP was restored by complementation of the deletion mutants with phoR-phoP. The biosynthesis of two secondary metabolites, actinorhodin and undecylprodigiosin, was significantly increased in both solid and liquid medium in the DeltaphoP or DeltaphoR-phoP deletion mutants. Negative phosphate control of both secondary metabolites was restored by complementation with the phoR-phoP cluster. These results prove that expression of both phoA and genes implicated in the biosynthesis of secondary metabolites in S. lividans is regulated by a mechanism involving the two component PhoR-PhoP system. PMID- 12730374 TI - Expanding tRNA recognition of a tRNA synthetase by a single amino acid change. AB - Aspartyl-tRNA synthetase (AspRS) occurs in two types: the discriminating enzyme (D-AspRS) forms only Asp-tRNA(Asp), whereas the nondiscriminating enzyme (ND AspRS) also synthesizes Asp-tRNA(Asn), which is a required intermediate for protein synthesis in many organisms. We attempted to expand the tRNA recognition of the discriminating Thermococcus kodakaraensis AspRS to that of a ND-AspRS by in vitro mutagenesis. An alignment of 26 archaeal AspRS proteins revealed two positions (26 and 85 in the T. kodakaraensis sequence) whose amino acid identity changes according to the enzymes' tRNA specificity. In their anticodon-binding domain, D-AspRS proteins contain W26 (or Q26) and K85, compared with H26 and P85 in the ND-AspRSs. T. kodakaraensis AspRS gained the ability to form Asp-tRNA(Asn) in vitro when the W26H or K85P changes were introduced independently or in combination. In the aminoacylation of tRNA(Asn) or tRNA(Asp) transcripts, the mutant enzymes displayed at least a 100- to 500-fold change in tRNA specificity, as judged by the ratio of the k(cat)K(m) values of Asp-tRNA(Asp) vs. Asp tRNA(Asn) formation. That T. kodakaraensis mutant AspRSs mischarge tRNA(Asn) was also manifested in the higher level (1.7%) of aspartylation of unfractionated Pyrococcus tRNA compared with that achieved by the wild-type enzyme (0.9%). Northern blot analysis of the Asp-tRNA separated by acidurea gel electrophoresis confirmed the in vitro synthesis of Asp-tRNA(Asn). A structure-based model points to a direct interaction of K85 in T. kodakaraensis AspRS with the anticodon nucleotide C36 of tRNA(Asp). Thus, a switch between D-AspRS and ND-AspRS enzymes could have evolved with only limited amino acid changes. PMID- 12730375 TI - Perturbations in B cell responsiveness to CD4+ T cell help in HIV-infected individuals. AB - HIV infection induces a wide array of B cell dysfunctions. We have characterized the effect of plasma viremia on the responsiveness of B cells to CD4(+) T cell help in HIV-infected patients. In HIV-negative donors, B cell proliferation correlated with CD154 expression on activated CD4(+) T cells and with the availability of IL-2, whereas in HIV-infected viremic patients, reduced B cell proliferation was observed despite normal CD154 expression on activated CD4(+) T cells. Reduced triggering of B cells by activated CD4(+) T cells was clearly observed in HIV-infected viremic patients compared with aviremic patients with comparable CD4(+) T cell counts, and a dramatic improvement in B cell function was observed in patients whose plasma viremia was controlled by effective antiretroviral therapy. The degree of B cell dysfunction in viremic patients correlated strongly with the inability of B cells to express CD25 in response to activated CD4(+) T cells, resulting in an inability to mount a normal proliferative response to IL-2. Similar defects in responsiveness to IL-2 were observed in the B cells of HIV-infected viremic patients in the context of B cell receptor stimulation. These data provide new insight into the mechanisms associated with ineffective humoral responses in HIV disease. PMID- 12730376 TI - The Na/K pump, Cl ion, and osmotic stabilization of cells. AB - An equation for membrane voltage is derived that takes into account the electrogenicity of the Na/K pump and is valid dynamically, as well as in the steady state. This equation is incorporated into a model for the osmotic stabilization of cells. The results emphasize the role of the pump and membrane voltage in lowering internal Cl(-) concentration, thus making osmotic room for vital substances that must be sequestered in the cell. PMID- 12730377 TI - Reverse engineering gene networks: integrating genetic perturbations with dynamical modeling. AB - While the fundamental building blocks of biology are being tabulated by the various genome projects, microarray technology is setting the stage for the task of deducing the connectivity of large-scale gene networks. We show how the perturbation of carefully chosen genes in a microarray experiment can be used in conjunction with a reverse engineering algorithm to reveal the architecture of an underlying gene regulatory network. Our iterative scheme identifies the network topology by analyzing the steady-state changes in gene expression resulting from the systematic perturbation of a particular node in the network. We highlight the validity of our reverse engineering approach through the successful deduction of the topology of a linear in numero gene network and a recently reported model for the segmentation polarity network in Drosophila melanogaster. Our method may prove useful in identifying and validating specific drug targets and in deconvolving the effects of chemical compounds. PMID- 12730378 TI - Positional cloning of the wheat vernalization gene VRN1. AB - Winter wheats require several weeks at low temperature to flower. This process, vernalization, is controlled mainly by the VRN1 gene. Using 6,190 gametes, we found VRN1 to be completely linked to MADS-box genes AP1 and AGLG1 in a 0.03 centimorgan interval flanked by genes Cysteine and Cytochrome B5. No additional genes were found between the last two genes in the 324-kb Triticum monococcum sequence or in the colinear regions in rice and sorghum. Wheat AP1 and AGLG1 genes were similar to Arabidopsis meristem identity genes AP1 and AGL2, respectively. AP1 transcription was regulated by vernalization in both apices and leaves, and the progressive increase of AP1 transcription was consistent with the progressive effect of vernalization on flowering time. Vernalization was required for AP1 transcription in apices and leaves in winter wheat but not in spring wheat. AGLG1 transcripts were detected during spike differentiation but not in vernalized apices or leaves, suggesting that AP1 acts upstream of AGLG1. No differences were detected between genotypes with different VRN1 alleles in the AP1 and AGLG1 coding regions, but three independent deletions were found in the promoter region of AP1. These results suggest that AP1 is a better candidate for VRN1 than AGLG1. The epistatic interactions between vernalization genes VRN1 and VRN2 suggested a model in which VRN2 would repress directly or indirectly the expression of AP1. A mutation in the promoter region of AP1 would result in the lack of recognition of the repressor and in a dominant spring growth habit. PMID- 12730380 TI - Controlling integration specificity of a yeast retrotransposon. AB - Retrotransposons and retroviruses integrate nonrandomly into eukaryotic genomes. For the yeast retrotransposon Ty5, integration preferentially occurs within domains of heterochromatin. Targeting to these locations is determined by interactions between an amino acid sequence motif at the C terminus of Ty5 integrase (IN) called the targeting domain, and the heterochromatin protein Sir4p. Here we show that new Ty5 integration hot spots are created when Sir4p is tethered to ectopic DNA sites. Targeting to sites of tethered Sir4p is abrogated by single amino acid substitutions in either IN or Sir4p that prevent their interaction. Ty5 target specificity can be altered by replacing the IN-targeting domain with other peptide motifs that interact with known protein partners. Integration occurs at high efficiency and in close proximity to DNA sites where the protein partners are tethered. These findings define a mechanism by which retrotransposons shape their host genomes and suggest ways in which retroviral integration can be controlled. PMID- 12730379 TI - Protein-protein interactions: structurally conserved residues distinguish between binding sites and exposed protein surfaces. AB - Polar residue hot spots have been observed at protein-protein binding sites. Here we show that hot spots occur predominantly at the interfaces of macromolecular complexes, distinguishing binding sites from the remainder of the surface. Consequently, hot spots can be used to define binding epitopes. We further show a correspondence between energy hot spots and structurally conserved residues. The number of structurally conserved residues, particularly of high ranking energy hot spots, increases with the binding site contact size. This finding may suggest that effectively dispersing hot spots within a large contact area, rather than compactly clustering them, may be a strategy to sustain essential key interactions while still allowing certain protein flexibility at the interface. Thus, most conserved polar residues at the binding interfaces confer rigidity to minimize the entropic cost on binding, whereas surrounding residues form a flexible cushion. Furthermore, our finding that similar residue hot spots occur across different protein families suggests that affinity and specificity are not necessarily coupled: higher affinity does not directly imply greater specificity. Conservation of Trp on the protein surface indicates a highly likely binding site. To a lesser extent, conservation of Phe and Met also imply a binding site. For all three residues, there is a significant conservation in binding sites, whereas there is no conservation on the exposed surface. A hybrid strategy, mapping sequence alignment onto a single structure illustrates the possibility of binding site identification around these three residues. PMID- 12730381 TI - Requirement for Pangolin/dTCF in Drosophila Wingless signaling. AB - The Wingless (Wg) protein is a secreted glycoprotein involved in intercellular signaling. On activation of the Wg signaling pathway, Armadillo is stabilized, causing target genes to be activated by the transcription factor Pangolin (Pan). This study investigated the roles of Pan in the developing wing of Drosophila by clonal analysis. Three different aspects of wing development were examined: cell proliferation, wing margin specification, and wg self-refinement. Our results indicate that Pan function is critically required for all three of these processes. Consequently, lack of pan causes a severe reduction in the activity of the Wg target genes Distalless and vestigial within their normal domain of expression. Loss of pan function does not, however, lead to a derepression of these genes outside this domain. Thus, although Pan is positively required for the induction of Wg targets in the wing imaginal disk, it does not appear to play a default repressor function in the absence of Wg input. In contrast, lack of zygotic pan function causes a milder phenotype than that caused by the lack of wg function in the embryo. We show that this difference cannot be attributed to maternally provided pan product, indicating that a Pan repressor function usually prevents the expression of embryonic Wg targets. Together, our results suggest that for embryonic patterning the activator as well as repressor forms of Pan play important roles, while for wing development Pan operates primarily in the activator mode. PMID- 12730385 TI - The threat of plant pathogens as weapons against U.S. crops. AB - The U.S. National Research Council (NRC) concluded in 2002 that U.S. agriculture is vulnerable to attack and that the country has inadequate plans for dealing with agricultural bioterrorism. This article addresses the vulnerability of U.S. crops to attack from biological weapons by reviewing the costs and impact of plant diseases on crops, pointing out the difficulty in preventing deliberate introduction of pathogens and discovering new disease outbreaks quickly, and discussing why a plant pathogen might be chosen as a biological weapon. To put the threat into context, a brief historical review of anti-crop biological weapons programs is given. The argument is made that the country can become much better prepared to counter bioterrorism by developing a list of likely anti-crop threat agents, or categories of agents, that is based on a formal risk analysis; making structural changes to the plant protection system, such as expanding diagnostic laboratories, networking the laboratories in a national system, and educating first responders; and by increasing our understanding of the molecular biology and epidemiology of threat agents, which could lead to improved disease control, faster and more sensitive diagnostic methods, and predictions of disease invasion, persistence, and spread following pathogen introduction. PMID- 12730383 TI - Distinct response to dioxin in an arylhydrocarbon receptor (AHR)-humanized mouse. AB - There are large inter- and intraspecies differences in susceptibility to dioxin induced toxicities. A critical question in risk assessment of dioxin and related compounds is whether humans are sensitive or resistant to their toxicities. The diverse responses of mammals to dioxin are strongly influenced by functional polymorphisms of the arylhydrocarbon receptor (AHR). To characterize responses mediated by the human AHR (hAHR), we generated a mouse possessing hAHR instead of mouse AHR. Responses of these mice to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and 3-methylcholanthrene were compared with the responses of naturally sensitive (C57BL6J) and resistant (DBA2) mice. Mice homozygous for hAHR exhibited weaker induction of AHR target genes such as cyp1a1 and cyp1a2 than did C57BL6J (Ahr(b 1/b-1)) mice. DBA2 (Ahr(d/d)) mice were less responsive to induction of cyp genes than C57BL6J mice. hAHR and DBA2 AHR exhibit similar ligand-binding affinities and homozygous hAHR and Ahr(d/d) mice displayed comparable induction of AHR target genes by 3-methylcholanthrene. However, when TCDD was administered, a greatly diminished response was observed in homozygous hAHR mice compared with Ahr(d/d) mice, indicating that hAHR expressed in mice is functionally less responsive to TCDD than DBA2 AHR. After maternal exposure to TCDD, homozygous hAHR fetuses developed embryonic hydronephrosis, but not cleft palate, whereas fetuses possessing Ahr(b-1) or Ahr(d) developed both anomalies. These results suggest that hAHR may define the specificity of the responses to various AHR ligands. Thus, the hAHR knock-in mouse is a humanized model mouse that may better predict the biological effects of bioaccumulative environmental toxicants like TCDD in humans. PMID- 12730382 TI - Substoichiometric shifting in the plant mitochondrial genome is influenced by a gene homologous to MutS. AB - The plant mitochondrial genome is retained in a multipartite structure that arises by a process of repeat-mediated homologous recombination. Low-frequency ectopic recombination also occurs, often producing sequence chimeras, aberrant ORFs, and novel subgenomic DNA molecules. This genomic plasticity may distinguish the plant mitochondrion from mammalian and fungal types. In plants, relative copy number of recombination-derived subgenomic DNA molecules within mitochondria is controlled by nuclear genes, and a genomic shifting process can result in their differential copy number suppression to nearly undetectable levels. We have cloned a nuclear gene that regulates mitochondrial substoichiometric shifting in Arabidopsis. The CHM gene was shown to encode a protein related to the MutS protein of Escherichia coli that is involved in mismatch repair and DNA recombination. We postulate that the process of substoichiometric shifting in plants may be a consequence of ectopic recombination suppression or replication stalling at ectopic recombination sites to effect molecule-specific copy number modulation. PMID- 12730386 TI - The potential of optical canopy measurement for targeted control of field crop diseases. AB - There is increasing pressure to reduce the use of pesticides in modern crop production to decrease the environmental impact of current practice and to lower production costs. It is therefore imperative that sprays are only applied when and where needed. Since diseases in fields are frequently patchy, sprays may be applied unnecessarily to disease-free areas. Disease control could be more efficient if disease patches within fields could be identified and spray applied only to the infected areas. Recent developments in optical sensor technology have the potential to enable direct detection of foliar disease under field conditions. This review assesses recent developments in the use of optical methods for detecting foliar disease, evaluates the likely benefits of spatially selective disease control in field crops, and discusses practicalities and limitations of using optical disease detection systems for crop protection in precision pest management. PMID- 12730387 TI - Spiroplasma citri, a plant pathogenic molligute: relationships with its two hosts, the plant and the leafhopper vector. AB - Spiroplasma citri, the type species of the genus Spiroplasma (Spiroplasmataceae, Mollicutes), is restricted to the phloem sieve tubes and transmitted by phloem sap-feeding insects, as is characteristic of the phytopathogenic mollicutes. The spiroplasmas are the only mollicutes showing motility and helical morphology, apparently mediated by a contractile fibrillar cytoskeleton bound to the inner surface of the spiroplasmal membrane. MreB genes, which are involved in cell shape determination, have been identified in S. citri. Identified genes of other functional groups are those involved in the transmission of S. citri by the leafhoppers and genes coding for lipoproteins, including spiralin, bound to the outer surface of the spiroplasma membrane. S. citri mutants that are unable to use fructose induce only mild and delayed symptoms. Fructose utilization by the sieve tube-restricted wild-type spiroplasmas is postulated to deprive the companion cells of fructose, thereby impairing sucrose loading into the sieve tubes. PMID- 12730384 TI - A cell-based assay for aggregation inhibitors as therapeutics of polyglutamine repeat disease and validation in Drosophila. AB - The formation of polyglutamine-containing aggregates and inclusions are hallmarks of pathogenesis in Huntington's disease that can be recapitulated in model systems. Although the contribution of inclusions to pathogenesis is unclear, cell based assays can be used to screen for chemical compounds that affect aggregation and may provide therapeutic benefit. We have developed inducible PC12 cell culture models to screen for loss of visible aggregates. To test the validity of this approach, compounds that inhibit aggregation in the PC12 cell-based screen were tested in a Drosophila model of polyglutamine-repeat disease. The disruption of aggregation in PC12 cells strongly correlates with suppression of neuronal degeneration in Drosophila. Thus, the engineered PC12 cells coupled with the Drosophila model provide a rapid and effective method to screen and validate compounds. PMID- 12730388 TI - Engineering plants for nematode resistance. AB - Biotechnology offers sustainable solutions to the problem of plant parasitic nematode control. There are several possible approaches for developing transgenic plants with improved nematode resistance; these include anti-invasion and migration strategies, feeding-cell attenuation, and antinematode feeding and development strategies. The essential elements of an effective control strategy are (a) genes that encode an antinematode effector protein, peptide or interfering RNA and (b) promoters that direct a specific pattern of expression for that effector. This review summarizes information on effectors that act directly against the nematode as well as those aimed at disrupting the nematode feeding site. We discuss patterns of promoter activity that could deliver expression of these effectors in a restricted and directed manner. Societal opposition to the technology of GM-nematode control is also discussed. PMID- 12730389 TI - Regulation of antibiotic production in root-colonizing Peudomonas spp. and relevance for biological control of plant disease. AB - Certain strains of fluorescent pseudomonads are important biological components of agricultural soils that are suppressive to diseases caused by pathogenic fungi on crop plants. The biocontrol abilities of such strains depend essentially on aggressive root colonization, induction of systemic resistance in the plant, and the production of diffusible or volatile antifungal antibiotics. Evidence that these compounds are produced in situ is based on their chemical extraction from the rhizosphere and on the expression of antibiotic biosynthetic genes in the producer strains colonizing plant roots. Well-characterized antibiotics with biocontrol properties include phenazines, 2,4-diacetylphloroglucinol, pyoluteorin, pyrrolnitrin, lipopeptides, and hydrogen cyanide. In vitro, optimal production of these compounds occurs at high cell densities and during conditions of restricted growth, involving (i) a number of transcriptional regulators, which are mostly pathway-specific, and (ii) the GacS/GacA two-component system, which globally exerts a positive effect on the production of extracellular metabolites at a posttranscriptional level. Small untranslated RNAs have important roles in the GacS/GacA signal transduction pathway. One challenge in future biocontrol research involves development of new strategies to overcome the broad toxicity and lack of antifungal specificity displayed by most biocontrol antibiotics studied so far. PMID- 12730390 TI - Quorum sensing in plant-pathogenic bacteria. AB - Quorum sensing (QS) allows bacteria to assess their local population density and/or physical confinement via the secretion and detection of small, diffusible signal molecules. This review describes how phytopathogenic bacteria have incorporated QS mechanisms into complex regulatory cascades that control genes for pathogenicity and colonization of host surfaces. Traits regulated by QS include the production of extracellular polysaccharides, degradative enzymes, antibiotics, siderophores, and pigments, as well as Hrp protein secretion, Ti plasmid transfer, motility, biofilm formation, and epiphytic fitness. Since QS regulatory systems are often required for pathogenesis, interference with QS signaling may offer a means of controlling bacterial diseases of plants. Several bacterial pathogens of plants that have been intensively studied and have revealed information of both fundamental and practical importance are reviewed here: Agrobacterium tumefaciens, Pantoea stewartii, Erwinia carotovora, Ralstonia solanacearum, Pseudomonas syringae, Pseudomonas aeruginosa, and Xanthomonas campestris. PMID- 12730391 TI - Perspectives on plant and soil nematology. AB - During my career in Plant Pathology/Nematology, many major advancements have occurred in the study of nematodes-even with their being largely soilborne and thus often overlooked. These biotrophic organisms include the most widespread and important group of plant pathogens-the root-knot nematodes Meloidogyne species which attack most major crops, as well as thousands of non-crop plant species. Landmark achievements that catalyzed research on these organisms included the discovery of effective nematicides, ectoparasitic forms, elucidation of disease complexes, nematodes as virus vectors, development of host resistance, and new technologies for research. Evolving research thrusts involve interfacing traditional and molecular systematics/diagnostics, adoption of the Caenorhabditis elegans-molecular genetics resource for general nematological research, focus on genetics of parasitism, use of molecular tools in developing host resistance, ecological and quantitative facets, and soil-biology-ecology based integrated management. Educational and international programs are encountering many changes and challenges, as is support for nematology in general. PMID- 12730392 TI - Pathogen self-defense: mechanisms to counteract microbial antagonism,. AB - Natural and agricultural ecosystems harbor a wide variety of microorganisms that play an integral role in plant health, crop productivity, and preservation of multiple ecosystem functions. Interactions within and among microbial communities are numerous and range from synergistic and mutualistic to antagonistic and parasitic. Antagonistic and parasitic interactions have been exploited in the area of biological control of plant pathogenic microorganisms. To date, biocontrol is typically viewed from the perspective of how antagonists affect pathogens. This review examines the other face of this interaction: how plant pathogens respond to antagonists and how this can affect the efficacy of biocontrol. Just as microbial antagonists utilize a diverse arsenal of mechanisms to dominate interactions with pathogens, pathogens have surprisingly diverse responses to counteract antagonism. These responses include detoxification, repression of biosynthetic genes involved in biocontrol, active efflux of antibiotics, and antibiotic resistance. Understanding pathogen self-defense mechanisms for coping with antagonist assault provides a novel approach to improving the durability of biologically based disease control strategies and has implications for the deployment of transgenes (microorganisms or plants). PMID- 12730393 TI - Evolution of Wheat streak mosaic virus: dynamics of population growth within plants may explain limited variation. AB - Like many other plant RNA viruses, Wheat streak mosaic virus (WSMV) sequence diversity within and among infected plants is low given the large number of virions produced. This may be explained by considering aspects of plant virus life history. Intracellular replication of RNA viruses is predominately linear, not exponential, which means that the rate at which mutations accumulate also is linear. Bottlenecks during systemic movement further limit diversity. Analysis of mixed infections with two WSMV isolates suggests that about four viral genomes participate in systemic invasion of each tiller. Low effective population size increases the role of stochastic processes on dynamics of plant virus population genetics and evolution. Despite low pair-wise diversity among isolates, the number of polymorphic sites within the U.S. population is about the same as between divergent strains or a sister species. Characteristics of polymorphism in the WSMV coat protein gene suggest that most variation appears neutral. PMID- 12730394 TI - James Gordon Horsfall: nonconformist and founding father. AB - James Gordon Horsfall led the creation of the Annual Review of Phytopathology. Preeminent in the invention of organic fungicides, he also starred in epidemiology. He employed his gift with words to encourage theory and practice to march together. PMID- 12730395 TI - Innovations in teaching plant pathology. AB - The teaching environment for plant pathology is changing in both positive and negative ways. Teaching expectations are increasing and resources are decreasing, but recent educational research and instructional technology offer new approaches to meet these challenges. Plant pathologists are teaching courses that may attract new students to the discipline or at least improve agricultural awareness. The Internet offers rapid access to information and images for both students and instructors. Instructional technology provides new tools for classroom presentations, communication with students, reaching new audiences, and distance learning, but using these new tools to enhance learning requires skilled and creative instructors. In the past, many plant pathology instructors worked in relative isolation, but new communication technologies and publishing opportunities for teaching scholarship should improve the sharing of instructional resources and methods. PMID- 12730396 TI - Ecology of mycorrhizae: a conceptual framework for complex interactions among plants and fungi. AB - Mycorrhizae regulate elemental and energy flows in terrestrial ecosystems. We understand much of how mycorrhizae work, but integrating all possible mechanisms into a whole has proven elusive. Multiple evolutionary events and the long evolutionary history mean that different plants and fungi bring independent characteristics to the symbiosis. This variety results in extensive physiological variation. How do we integrate functional responses with diversity to understand the role of mycorrhizae in ecosystems? We review ecophysiological mechanisms of mycorrhizae and organize these into functional groups. Species-area relationships are not curvilinear, but resemble the "broken stick" model. We coupled functional groups with a metacommunity analysis to show how complex behavior can be generated using a simple matrix model of resource exchange. This approach provides insights into how we might integrate diversity and function across landscapes. PMID- 12730397 TI - Cultural and genetic approaches to managing mycotoxins in maize. AB - Infection of maize kernels by toxigenic fungi remains a challenging problem despite decades of research progress. Cultural practices, including crop rotation, tillage, planting date, and management of irrigation and fertilization, have limited effects on infection and subsequent mycotoxin accumulation. Current infrastructure and grain storage practices in developed countries can prevent postharvest development of mycotoxins, but this aspect remains a threat in developing countries, especially in tropical areas. Because most mycotoxin problems develop in the field, strategies are needed to prevent infection of growing plants by toxigenic fungi. Developing genetic resistance to Aspergillus flavus, Gibberella zeae, and Fusarium spp. (particularly F. verticillioides) in maize is a high priority. Sources of resistance to each of these pathogens have been identified and have been incorporated into public and private breeding programs. However, few, if any, commercial cultivars have adequate levels of resistance. Efforts to control infection or mycotoxin development through conventional breeding and genetic engineering are reviewed. The role of transgenic insect control in the prevention of mycotoxins in maize is discussed. PMID- 12730398 TI - Carl Freiherr von Tubeuf: pioneer in biological control of plant diseases. AB - Carl von Tubeuf, a founder of plant pathology in Europe, pioneered biological control of a major plant disease and introduced the term "biological control" to plant pathology. His book on plant diseases was one of the first to be published in English. He was proficient in many areas, and his interests included plant diseases, mycology, forest insects, white pine blister rust, the mistletoes, plant protection laws, and nature conservation. PMID- 12730399 TI - The ecological significance of biofilm formation by plant-associated bacteria. AB - Bacteria associated with plants have been observed frequently to form assemblages referred to as aggregates, microcolonies, symplasmata, or biofilms on leaves and on root surfaces and within intercellular spaces of plant tissues. In a wide range of habitats, biofilms are purported to be microniches of conditions markedly different from those of the ambient environment and drive microbial cells to effect functions not possible alone or outside of biofilms. This review constructs a portrait of how biofilms associated with leaves, roots and within intercellular spaces influence the ecology of the bacteria they harbor and the relationship of bacteria with plants. We also consider how biofilms may enhance airborne dissemination, ubiquity and diversification of plant-associated bacteria and may influence strategies for biological control of plant disease and for assuring food safety. Trapped by a nexus, coordinates uncertain Ever expanding or contracting Cannibalistic and scavenging sorties Excavations through signs of past alliances Consensus signals sound revelry Then time warped by viscosity Genomes showing codependence A virtual microbial beach party With no curfew and no time-out A few estranged cells seeking exit options, Looking for another menagerie. David Sands, Montana State University, Bozeman, February 2003 PMID- 12730400 TI - Luteovirus-aphid interactions. AB - Members of the Luteoviridae are transmitted by aphids in a circulative, nonpropagative manner that requires the virus to be acquired through gut tissue into the aphid hemocoel and then exit through salivary tissues. This process is aphid species-specific and involves specific recognition of the virus by unidentified components on the membranes of gut and salivary tissues. Transport through the tissues is an endocytosis/exocytosis process. Both structural proteins of the virus are involved in the transmission process, with multiple protein domains regulating the movement and survival of the virus in the aphid and plant. Here we review what is known about the genetic, cellular, and molecular mechanisms regulating these complex and specific virus-aphid interactions. PMID- 12730401 TI - Proceedings of the 11th International Symposium on Trace Elements in Man and Animals. Berkeley, California. June 2-6, 2002. PMID- 12730402 TI - Environmental contamination and chronic inflammation influence human growth potential. PMID- 12730404 TI - Dietary soy protein maintains some indices of bone mineral density and bone formation in aged ovariectomized rats. AB - Hormone replacement therapy (HRT) has been used to prevent osteoporosis in postmenopausal women. However, HRT may increase the incidence of some cancers and has other side effects. There is considerable interest in dietary alternatives that include the consumption of soy and isoflavones derived from soy. The purpose of this study was to determine the effects of dietary soy protein on bone density, formation and resorption in cortical and cancellous bone in aged, ovariectomized rats. Specific emphasis was placed on indices of bone formation. Rats were assigned to the following groups: baseline; sham surgery + casein diet; sham + soy protein diet; ovariectomy (Ovx) + casein diet; Ovx + soy protein diet. The diets were fed for 3 mo. The Ovx Soy group had a greater bone mineral density (BMD) than the Ovx Casein group. There was a trend (P < 0.10) for greater periosteal bone formation rates in the Sham Soy compared with the Sham Casein group. In the Ovx Soy group, indices of endocortical bone formation were greater than those of the Ovx Casein group. There were no significant differences in resorption indices or endochondral growth (bone elongation) rates with soy in either the Sham or Ovx groups. In cancellous bone, the double-labeled surface and bone formation rates were greater in the Ovx Soy group than in the Ovx Casein group. These results show that dietary soy had a beneficial effect on the preservation of BMD associated with estrogen deficiency bone loss in aged rats. These data also show that at the tissue level, soy functions in a manner different from estrogen by increasing or sustaining elevated bone formation rates after ovariectomy. PMID- 12730405 TI - Soy protein isolate enhances hepatic copper accumulation and cell damage in LEC rats. AB - In a series of experiments, the effects of soy protein isolate (SPI), defatted soy (DFS) or SPI supplemented with L-methionine (SPIM) were examined in the Long Evans rat with a cinnamon coat color (LEC rat), a model animal of Wilson's disease with a hereditary defect in the Atp7b gene resulting in defective copper metabolism and copper accumulation in hepatocytes. Milk casein in the control AIN 93G diet (20 g/100 g) was totally or 60% replaced by the soy products, SPI, DFS or SPIM (L-Met added to be equal to that in the control diet) beginning when rats were 6 wk old. Copper and iron concentrations in SPI and DFS were measured and the concentrations of these metals in the salt mix were adjusted so that test and the control diets had the same final concentrations. Food intake did not differ among groups. Rats were euthanized when they became moribund with jaundice. Survival time in the SPI diet group was shorter (14.0 +/- 0.8 wk) than in the control group (19.1 +/- 1.7 wk) (P < 0.001), and that in the DFS diet group was intermediate (16.0 +/- 1.7 wk). Survival time in the SPIM diet group did not differ from that of the SPI diet group. Copper concentrations in the livers of rats in the SPI and SPIM diet groups were approximately 80% higher than in rats fed the control diet. Liver iron concentrations did not differ among the groups. The results, including histological analyses, indicate that SPI enhances copper uptake into the liver cells and promotes liver cell damage in LEC rats. However, this did not occur in the livers of F344 rats with wild-type Atp7b. Recommendations to individuals suffering from Wilson's disease to avoid consuming soy protein may be warranted. PMID- 12730403 TI - Soy isoflavones exert antidiabetic and hypolipidemic effects through the PPAR pathways in obese Zucker rats and murine RAW 264.7 cells. AB - The hypocholesterolemic and anti-atherosclerotic mechanism by which soy may exert a beneficial effect remains unclear. Peroxisome-proliferator activated receptors (PPAR) are promiscuous nuclear receptors that regulate the transcription of genes involved in lipid and glucose homeostasis and lipid metabolism within the cell. We hypothesize that the isoflavones improve lipid and glucose metabolism by acting as an antidiabetic PPAR agonist. Male and female obese Zucker rats (OZR) were used as a model of Type 2 diabetes, and OZR fed a high isoflavone soy protein diet displayed improvements in lipid metabolism consistent with results in humans treated with antidiabetic PPAR agonists such as the fibrates or glitazones. Liver triglyceride and cholesterol concentrations were lower in all OZR fed high-isoflavone soy protein diets than in rats fed low-isoflavone and casein diets (P < 0.05). Concurrently, PPAR-directed gene expression was evaluated in a cell culture model. An isoflavone-containing soy extract doubled PPAR-directed gene expression (P < 0.05) in RAW 264.7 cells containing either a PPARalpha or PPARgamma expression plasmid. A similar induction was observed when the soy isoflavones genistein or daidzein were used to treat cells. Both isoflavones doubled PPARalpha-directed gene expression (P < 0.05), whereas they increased PPARgamma-directed gene expression 200-400% (P < 0.05). This study suggests that soy isoflavones improve lipid metabolism, produce an antidiabetic effect, and activate PPAR receptors. PMID- 12730406 TI - The addition of soybean phosphatidylcholine to triglyceride increases suppressive effects on food intake and gastric emptying in rats. AB - The physiologic roles of dietary lecithin have not yet been clearly defined. We examined the effects of soybean lecithin on gastric emptying (Experiments 1 and 2) and food intake (Experiment 3) in rats. Male Wistar rats were fed 2 g of a 20 g lipid/100 g diet containing various levels of lecithin after 24 h of food deprivation; gastric contents were collected 3 h after feeding (Experiment 1). In Experiment 2, the effects of lecithin and a CCK-A receptor antagonist on gastric emptying were examined using a modified phenol red recovery technique. In Experiment 3, their effects on food intake were examined after an intraduodenal infusion of an oil emulsion containing 50 mg soybean oil (SO) or SO partially replaced by lecithin (14-50%). Gastric emptying rates of the lipid and protein in the test diet (Experiment 1) or of phenol red (Experiment 2) were lower in the groups administered lecithin. Food intake for 60 min was lower in rats infused with the oil emulsion containing lecithin (25, 50%) than in rats not administered lecithin. The suppressive effects of lecithin on gastric emptying and food intake were largely reduced by devazepide. These results demonstrate that oil containing lecithin inhibits gastric emptying and food intake, and the effects are associated in part with CCK release. PMID- 12730407 TI - Diaminobiotin and desthiobiotin have biotin-like activities in Jurkat cells. AB - In mammals, biotin serves as a coenzyme for carboxylases such as propionyl-CoA carboxylase. The expression of genes encoding interleukin-2 (IL-2) and IL-2 receptor (IL-2R)gamma also depends on biotin. Biotin metabolites are structurally similar to biotin, and their concentrations in tissues are quantitatively important. Here, the hypothesis was tested that biotin metabolites can mimic the effects of biotin on gene expression and thus have biotin-like activities. A human T-cell line (Jurkat cells) was used to model effects of biotin and synthetic metabolites (diaminobiotin and desthiobiotin) on the expression of genes encoding IL-2 and IL-2Rgamma. Cells were cultured in biotin-deficient medium (0.025 nmol/L biotin) for 35 d; controls were cultured in medium containing 10 nmol/L biotin. The biotin-deficient medium was supplemented with 10 nmol/L of diaminobiotin, desthiobiotin, biotin or no biotin 24 h before gene expression analyses. Transcriptional activities of genes encoding IL-2 and IL 2Rgamma were increased up to 43% in cells supplemented with diaminobiotin, desthiobiotin or biotin compared with biotin-deficient cells, as judged by luciferase activities after transfection with reporter-gene constructs. These findings are consistent with the hypothesis that diaminobiotin and desthiobiotin mimic the effects of biotin on gene expression and thus have biotin-like activities. Supplementation of cells with diaminobiotin and desthiobiotin did not affect abundances of holocarboxylases and activities of propionyl-CoA carboxylase, suggesting that effects of synthetic biotin metabolites on gene expression are not mediated by carboxylase-dependent pathways. It is not known whether naturally occurring biotin metabolites also have biotin-like activities. PMID- 12730408 TI - New leptin receptor mutations in mice: Lepr(db-rtnd), Lepr(db-dmpg) and Lepr(db rlpy). AB - Three new spontaneous recessive mouse mutations in the leptin receptor gene (Lepr), Lepr(db-rtnd), Lepr(db-dmpg) and Lepr(db-rlpy), originated in the CBA/J (CBA), B10.D2-H8(b)(57N)/Sn (B10) and NU/J strains, respectively. Lepr(db-rtnd) and Lepr(db-dmpg) were maintained on C57BL/6J (B6), resulting in congenic lines of B6.CBA-Lepr(db-rtnd) and B6.B10-Lepr(db-dmpg). Lepr(db-rtnd) was also maintained on CBA post F1 generation of a cross between the B6 and the CBA, generating the congenic line CBA.B6CBA-Lepr(db-rtnd). Lepr(db-rlpy) was maintained as a coisogenic strain. The aims of this study were to determine the molecular bases for these new Lepr mutations and to characterize the new mutant stocks, with respect to obesity and diabetes. Mutations were analyzed by Southern blot analysis, reverse transcriptase-polymerase chain reaction and sequencing. Body weights and plasma glucose and insulin levels were measured, and the histology of the pancreas was carried out. Lepr(db-rtnd) contained one G deletion in exon 4 of Lepr, introducing a frameshift and premature termination. Lepr(db dmpg) had a deletion in the extracellular domain of LEPR: Lepr(db-rlpy) exhibited a large DNA deletion, leading to a complete lack of LEPR: All three mutations led to morbid obesity and diabetes. It is noteworthy that Lepr(db-rtnd) caused milder hyperglycemia accompanied by higher plasma and pancreatic insulin contents on B6 compared to that on CBA backgrounds. In summary, we discovered three new mutations of Lepr, providing new mouse models for obesity and diabetes. Furthermore, our mutant stocks will be useful in elucidating the effects of the genetic background on the Lepr mutations and in testing the specificity of antibodies to LEPR. PMID- 12730409 TI - Methylenetetrahydrofolate reductase 677C-->T variant modulates folate status response to controlled folate intakes in young women. AB - A common genetic variant in the methylenetetrahydrofolate reductase (MTHFR) gene involving a cytosine to thymidine (C-->T) transition at nucleotide 677 is associated with reduced enzyme activity, altered folate status and potentially higher folate requirements. The objectives of this study were to investigate the effect of the MTHFR 677 T allele on folate status variables in Mexican women (n = 43; 18-45 y) and to assess the adequacy of the 1998 folate U.S. Recommended Dietary Allowance (RDA), 400 micro g/d as dietary folate equivalents (DFE). Subjects (14 CC, 12 CT, 17 TT genotypes) consumed a low folate diet (135 micro g/d DFE) for 7 wk followed by repletion with 400 micro g/d DFE (7 CC, 6 CT, 9 TT) or 800 micro g/d DFE (7 CC, 6 CT, 8 TT) for 7 wk. Throughout repletion with 400 micro g/d DFE, the TT genotype had lower (P 0.05) in their response relative to the CC genotype. Throughout repletion with 800 micro g/d DFE, the CT genotype had lower (P 0.05) in the measured variables between the TT and CC genotypes. Repletion with 400 micro g/d DFE led to normal blood folate and desirable plasma tHcy concentrations, regardless of MTHFR C677T genotype. Collectively, these data demonstrate that the MTHFR C-->T variant modulates folate status response to controlled folate intakes and support the adequacy of the 1998 folate U.S. RDA for all three MTHFR C677T genotypes. PMID- 12730411 TI - Several culinary and medicinal herbs are important sources of dietary antioxidants. AB - We assessed the contribution of culinary and medicinal herbs to the total intake of dietary antioxidants. Our results demonstrate that there is more than a 1000 fold difference among antioxidant concentrations of various herbs. Of the dried culinary herbs tested, oregano, sage, peppermint, garden thyme, lemon balm, clove, allspice and cinnamon as well as the Chinese medicinal herbs Cinnamomi cortex and Scutellariae radix all contained very high concentrations of antioxidants (i.e., >75 mmol/100 g). In a normal diet, intake of herbs may therefore contribute significantly to the total intake of plant antioxidants, and be an even better source of dietary antioxidants than many other food groups such as fruits, berries, cereals and vegetables. In addition, the herbal drug, Stronger Neo-Minophagen C, a glycyrrhizin preparation used as an intravenous injection for the treatment of chronic hepatitis, boosts total antioxidant intake. It is tempting to speculate that several of the effects due to these herbs are mediated by their antioxidant activities. PMID- 12730410 TI - The interaction between MTHFR 677 C-->T genotype and folate status is a determinant of coronary atherosclerosis risk. AB - The 677 C-->T polymorphism in the 5,10-methylenetetrahydrofolate reductase (MTHFR) gene interacts with folate status in determining elevated total plasma levels of homocysteine, a risk factor for coronary atherosclerotic disease (CAD). The present study had the following goals: 1) to define the 677 C-->T genotype specific threshold values of both plasma and RBC folate, associated with hyperhomocysteinemia (>15 micro mol/L); and 2) to determine the risk of CAD among subjects with levels of folate below the genotype-specific threshold considered at risk for hyperhomocysteinemia. We examined 655 subjects, with (433) or without (222) angiographically documented CAD. The MTHFR 677 C-->T genotype-specific threshold values of plasma folate corresponded to the 40th, 30th and 10th percentile in the TT, CT and CC genotype, respectively. A multivariate logistic regression analysis showed that the risk of CAD among subjects with plasma folate levels below the genotype-specific thresholds was 1.6 (95% CI, 1.04-2.46). Similar results were obtained when RBC folate was considered as a measure of folate status (odds ratio = 1.8, 95% CI, 1.03-3.15). A gene-nutrient interaction that defines a higher risk for CAD is determined by folate levels below specific thresholds, which differ depending on the MTHFR 677 C-->T genotype. PMID- 12730412 TI - Betaine supplementation lowers plasma homocysteine in healthy men and women. AB - Elevated levels of plasma total homocysteine are associated with a higher risk of cardiovascular disease. Betaine and 5-methyltetrahydrofolate can remethylate homocysteine into methionine via independent reactions. We determined the effect of daily betaine supplementation, compared with both folic acid and placebo, on plasma concentrations of total homocysteine after an overnight fast and after methionine loading in men and women with mildly elevated homocysteine. Groups of twelve subjects ingested 6 g betaine, 800 micro g folic acid with 6 g placebo or 6 g placebo each day for 6 wk. A methionine-loading test (i.e., ingestion of 100 mg L-methionine/kg body mass) was performed before and after 6 wk of supplementation. Fasting plasma homocysteine decreased by 1.8 micro mol/L (95% confidence interval [CI]: -3.6, 0.0, P < 0.05) in the betaine group and by 2.7 micro mol/L (95% CI: -4.5, -0.9, P < 0.05) in the folic acid group. These changes are relative to the change in the placebo group, in which fasting plasma homocysteine rose by 0.5 micro mol/L. Furthermore, betaine suppressed the total area under the plasma homocysteine-time curve after methionine loading by 221 micro mol. 24 h/L (95% CI: -425, -16, P < 0.05) compared with placebo, whereas folic acid had no effect. In conclusion, betaine appears to be highly effective in preventing a rise in plasma homocysteine concentration after methionine intake in subjects with mildly elevated homocysteine. It is not known whether this potential of betaine to "stabilize" circulating homocysteine concentrations lowers the risk of cardiovascular disease. PMID- 12730413 TI - Strawberry anthocyanins are recovered in urine as glucuro- and sulfoconjugates in humans. AB - Anthocyanins are phenolic compounds widely distributed in fruits and vegetables. Their consumption has been shown to prevent some chronic diseases. Anthocyanin metabolism, however, is still not fully understood. The aim of this work was to evaluate the bioavailability of anthocyanins in humans consuming a meal containing strawberries and to identify possible metabolites in urine. Six healthy volunteers (three women and three men) consumed a meal containing 200 g strawberries (providing 179 micro mol pelargonidin-3-glucoside). Urine samples were collected before and after the meal and rapidly treated by solid-phase extraction. Identification and quantification of anthocyanin metabolites were carried out by HPLC-ESI-MS-MS and HPLC with UV-visible detection, respectively. In addition to pelargonidin-3-glucoside, five anthocyanin metabolites were identified in urine: three monoglucuronides of pelargonidin, one sulfoconjugate of pelargonidin and pelargonidin itself. Total urinary excretion of strawberry anthocyanin metabolites corresponded to 1.80 +/- 0.29% (mean +/- SEM, n = 6) of pelargonidin-3-glucoside ingested. More than 80% of this excretion was related to a monoglucuronide. Four hours after the meal, more than two-thirds of anthocyanin metabolites had been excreted, although urinary excretion of the metabolites continued until the end of the 24-h experiment. This study demonstrated that anthocyanins were glucuro- and sulfo-conjugated in humans and that the main metabolite of strawberry anthocyanins in human urine was a monoglucuronide of pelargonidin. PMID- 12730414 TI - Concentrations of choline-containing compounds and betaine in common foods. AB - Choline is important for normal membrane function, acetylcholine synthesis and methyl group metabolism; the choline requirement for humans is 550 mg/d for men (Adequate Intake). Betaine, a choline derivative, is important because of its role in the donation of methyl groups to homocysteine to form methionine. In tissues and foods, there are multiple choline compounds that contribute to total choline concentration (choline, glycerophosphocholine, phosphocholine, phosphatidylcholine and sphingomyelin). In this study, we collected representative food samples and analyzed the choline concentration of 145 common foods using liquid chromatography-mass spectrometry. Foods with the highest total choline concentration (mg/100 g) were: beef liver (418), chicken liver (290), eggs (251), wheat germ (152), bacon (125), dried soybeans (116) and pork (103). The foods with the highest betaine concentration (mg/100 g) were: wheat bran (1339), wheat germ (1241), spinach (645), pretzels (237), shrimp (218) and wheat bread (201). A number of epidemiologic studies have examined the relationship between dietary folic acid and cancer or heart disease. It may be helpful to also consider choline intake as a confounding factor because folate and choline methyl donation can be interchangeable. PMID- 12730415 TI - Postprandial kinetics of dietary amino acids are the main determinant of their metabolism after soy or milk protein ingestion in humans. AB - Soy proteins have been shown to result in lower postprandial nitrogen retention than milk proteins, but the mechanisms underlying these differences have not been elucidated. To investigate this question, we measured the postprandial kinetics of the appearance of individual (15)N-amino acids in the serum of healthy adults after the ingestion of either (15)N-soy (n = 8) or (15)N-milk proteins (n = 8) in a mixed single meal (46 kJ/kg). The kinetics of total and dietary amino acids (AA) in the peripheral circulation were characterized by an earlier and higher peak after soy protein ingestion. Dietary AA levels peaked at 2.5 h in the soy group vs. 3.9 h in the milk group (P < 0.02). This time interval difference between groups was associated with a faster transfer of dietary N into urea in the soy group (peak at 3 vs. 4.75 h in the milk group, P < 0.005) and a higher level of incorporation into the serum protein pool from 3 to 8 h after the soy meal. The dietary AA pattern in the peripheral blood closely reflected the dietary protein AA pattern. Postprandial glucose, insulin, and glucagon levels and profiles did not differ between groups. Soy AA were digested more rapidly and were directed toward both deamination pathways and liver protein synthesis more than milk AA. We conclude that differences in the metabolic postprandial fates of soy and milk proteins are due mainly to differences in digestion kinetics; however, the AA composition of dietary proteins may also play a role. PMID- 12730416 TI - Soy isoflavones do not modulate circulating insulin-like growth factor concentrations in an older population in an intervention trial. AB - Insulin-like growth factors (IGF) are essential for normal growth and maintenance of lean muscle mass; however, high insulin-like growth factor-I (IGF-I) and low IGF binding protein-3 (IGFBP-3) levels are also associated with several cancers. To test the hypothesis that long-term soy isoflavone supplementation decreases circulating IGF-I concentrations, we conducted a controlled, parallel-arm, double blind intervention study with 150 participants (85% men), 50-80 y old. Participants were randomly assigned to consume a soy beverage powder daily for 12 mo. The active treatment group (+ISO) received soy protein containing 83 mg isoflavones, whereas the comparison group (-ISO) received soy protein containing 3 mg isoflavones. Serum IGF-I and IGFBP-3 were measured by ELISA. Mean change in serum IGF-I concentrations was similar in the two groups (+1.4 nmol/L in +ISO, +1.2 nmol/L in -ISO; P = 0.74, 95% confidence interval -1.1, +1.5 nmol/L for the 0.21 nmol/L difference between groups), indicating no effect of the isoflavone intervention. Similarly, the changes in IGFBP-3 and the IGF-I/IGFBP-3 ratio were similar in both groups, again showing no effect of +ISO treatment. A 12 mo, 83 mg/d soy isoflavone intervention did not modulate serum IGF in an older, mostly male population. PMID- 12730417 TI - Association of household rice expenditure with child nutritional status indicates a role for macroeconomic food policy in combating malnutrition. AB - Macroeconomic food policies have the potential to reduce malnutrition by improving access to food, a determinant of nutritional status. However, very little is understood about the mechanisms and the magnitude of the effects of macroeconomic food policies such as food price policies on nutritional status. Data collected by the Nutritional Surveillance Project on a total of 81,337 children aged 6-59 mo in rural Bangladesh between 1992 and 2000 were used to examine how changes in rice price affect child underweight. Rice consumption per capita declined only slightly during the period but rice expenditure per capita varied widely due to fluctuations in rice price. Rice expenditure was positively correlated with the percentage of underweight children (r = 0.91, P = 0.001). Households were found to spend more on nonrice foods as their rice expenditure declined, and nonrice expenditure per capita was negatively correlated with the percentage of underweight children (r = -0.91, P = 0.001). Expenditure on nonrice foods per capita increased with the frequency with which nonrice foods were consumed (P < 0.05) and with the diversity of the diet (P < 0.001). The findings suggest that the percentage of underweight children declined when rice expenditure fell because households were able to spend more on nonrice foods and thereby increase the quantity and quality of their diet. We hypothesize that macroeconomic food policies that keep the price of food staples low can contribute toward reducing child underweight. PMID- 12730418 TI - Infant feeding practices of HIV-positive mothers in India. AB - Exclusive breast-feeding is widely accepted and advocated in India; however, clinicians are now faced with advising women infected with human immunodeficiency virus (HIV) about the risks and benefits of other infant feeding options. This study assessed factors that influence the infant feeding decisions of HIV infected mothers in Pune, India. From December 2000 to April 2002, HIV-positive (HIV(+)) pregnant women (n = 101) from a government hospital antenatal clinic were interviewed prepartum about infant feeding intention, feeding practice immediately postpartum and feeding after a minimum of 2 wk postpartum. Of the HIV(+) sample, the last 39 were interviewed more intensively to examine factors affecting feeding decision making. We found that an equal number of HIV(+) women intended to breast-feed (44%) or give top milk (44%) (diluted animal milk). Women who chose to top feed were also more likely to disclose their HIV status to family members. Mixed feeding occurred frequently in our sample (29%); however, for the majority of those (74%), it lasted only 3 d postpartum. The hospital counselor had an important role in assisting women in their intended feeding choice as well as actual practice. The time immediately after delivery was noted as critical for recounseling about infant feeding and further support of the woman's decision, thus lowering the risk of mixed feeding. Lack of funds, poor hygienic conditions and risk of social repercussions were more commonly noted as reasons to breast-feed. Top milk, the alternative for breast-milk used in this population, however, must be investigated further to assess its nutritional value and safety before it can be endorsed widely for infants of HIV(+) women. PMID- 12730419 TI - Growth faltering in rural Gambian infants is associated with impaired small intestinal barrier function, leading to endotoxemia and systemic inflammation. AB - Growth faltering of rural Gambian infants is associated with a chronic inflammatory enteropathy of the mucosa of the small intestine that may impair both digestive/absorptive and barrier functions. The aim of this study was to determine whether the enteropathy was associated with a compromised barrier function that allowed translocation of antigenic macromolecules from the gut lumen into the body, with subsequent systemic immunostimulation, resulting in growth retardation. Rural Gambian infants were studied longitudinally at regular intervals between 8 and 64 wk of age. On each study day, each child was medically examined, anthropometric measurements were made, a blood sample was taken and an intestinal permeability test performed. Evidence of chronic immunostimulation was provided by abnormally elevated white blood cell, lymphocyte and platelet counts, and frequently raised plasma concentration of C-reactive protein. Intestinal permeability was abnormal and associated with impaired growth (r = -0.41, P < 0.001). Plasma concentrations of endotoxin and immunoglobulin (Ig)G-endotoxin core antibody were also elevated and related to both growth (r = -0.30, P < 0.02; r = -0.64, P < 0.0001, respectively) and measures of mucosal enteropathy. Plasma IgG, IgA and IgM levels increased rapidly with age toward adult concentrations. Raised values were related to poor growth but also to measures of mucosal enteropathy and the endotoxin antibody titer. The interrelationships among these variables and growth suggested that they were all part of the same growth retarding mechanism. These data are consistent with the hypothesis of translocation of immunogenic lumenal macromolecules across a compromised gut mucosa, leading to stimulation of systemic immune/inflammatory processes and subsequent growth impairment. PMID- 12730420 TI - A micronutrient-fortified beverage prevents iron deficiency, reduces anemia and improves the hemoglobin concentration of pregnant Tanzanian women. AB - Maternal malnutrition continues to be a major contributor to adverse reproductive outcomes in developing countries, despite longstanding efforts to fortify foods or to distribute medicinal supplements to pregnant women. The objective of this study was to test the effect of a micronutrient-fortified beverage containing 11 micronutrients (iron, iodine, zinc, vitamin A, vitamin C, niacin, riboflavin, folate, vitamin B-12, vitamin B-6 and vitamin E) on the hemoglobin, iron and vitamin A status of pregnant women in Tanzania. A group of 259 pregnant women with gestational ages of 8 to 34 wk were enrolled in a randomized double-blind controlled trial in which study women received 8 wk of supplementation. Hemoglobin, ferritin and dried blood spot retinol were measured at baseline and at the end of the supplementation period. The supplement resulted in a 4.16 g/L increase in hemoglobin concentration and a 3 micro g/L increase in ferritin and reduced the risk of anemia and iron deficiency anemia by 51 and 56%, respectively. The risk of iron deficiency was reduced by 70% among those who had iron deficiency at baseline and by 92% among those who had adequate stores. The micronutrient-fortified beverage may be a useful and convenient preventative measure, one that could help improve the nutritional status of women both before and during pregnancy and thereby help avoid some of the potential maternal and fetal consequences of micronutrient deficiencies. PMID- 12730422 TI - Ferulic acid sugar esters are recovered in rat plasma and urine mainly as the sulfoglucuronide of ferulic acid. AB - Ferulic acid sugar esters, the common form of ferulic acid (FA) in cereals, show a stronger antioxidant potential than FA in vitro. However, there is little information on their metabolism and excretion in vivo. In the present study, we investigated the metabolic derivatives of FA in the plasma, urine and feces of rats administered 70 micro mol/kg body of 5-O-feruloyl-L-arabinofuranose (FAA), feruloyl-arabinoxylan (FAXn) or the same molar amount of FA as a comparison. Administered FA and its sugar esters were recovered in rat plasma and urine in the form of free FA, FA-glucuronide, FA-sulfate and FA-sulfoglucuronide (FA diconjugate with sulfate and glucuronide). The recovery of administered FA in urine was 72%, which was higher than that of administered FAA (54%) or FAXn (20%). Free FA and its derivatives were not recovered in rat feces after FA or FAA administration, but 20% of the administered FA moiety was recovered when FAXn was administered. Moreover, administered FA, in contrast to FA esters, was present in plasma in the free and conjugated forms at a higher concentration but for a shorter time. These results indicated that bioavailability of FA and its sugar esters is dependent on the absence or presence of the saccharide moiety and, in the latter case, its structure. This is the first study to show that FA sulfoglucuronide is the main metabolite (60-70% of the total) in the plasma of rats administered FA or its sugar esters. Thus, the physiological functions of FA and its sugar esters found in vitro might require reconsideration in vivo. PMID- 12730421 TI - Folate derived from cecal bacterial fermentation does not increase liver folate stores in 28-d folate-depleted male Sprague-Dawley rats. AB - This study assessed the ability of rats to absorb and store the folate synthesized by cecal bacteria. Male weanling Sprague-Dawley rats were folate depleted by feeding a low folacin AIN93G formulated basal diet for 28 d; they were then fed repletion diets containing folate (0.25-1.0 mg/kg diet), dietary fiber (DF; wheat bran, oat bran, ground corn, wheat germ) or undigested and fermented dietary material (UFDM; polydextrose, inulin) in the presence and absence of an antibiotic (succinylsulfathiazole). Fermentation was stimulated by DF and UFDM and reduced by the antibiotic. In the absence of succinylsulfathiazole, the increase in liver folate (during the repletion phase) was proportional only to the folate content of the diet and did not vary with added DF or UFDM. Adding succinylsulfathiazole lowered total folate excretion from 13.8 +/- 8.2 to 4.8 +/- 2.9 nmol/d (pooled diets, P < 0.00001) in agreement with its role in inhibiting bacterial folate synthesis. In addition, succinylsulfathiazole lowered liver folate in rats fed control and test diets approximately equally with a mean decrease from 11.6 +/- 2.5 to 7.5 +/- 2.5 nmol/g wet liver (pooled diets, P < 0.00001), suggesting that the antibiotic also affected rat folate absorption and/or metabolism. Increased bacterial fermentation and excretion as well as increased bacterial folate production in the presence of added DF and UFDM were demonstrated by increased volatile fatty acid content in cecal and fecal samples (P < 0.000001) and increased diaminopimelic acid, muramic acid and folate in feces (P < 0.00001). The magnitude of these changes depended on the type of DF and UFDM. These results show that bacterially synthesized folate is not substantially absorbed and stored in the liver of Sprague-Dawley male rats. PMID- 12730423 TI - Hydrolytic activity toward pyridoxine-5'-beta-D-glucoside in rat intestinal mucosa is not increased by vitamin B-6 deficiency: effect of basal diet composition and pyridoxine intake. AB - Pyridoxine-5'-beta-D-glucoside (PNG), a glycosylated form of dietary vitamin B-6, is partially hydrolyzed in the small intestine by the cytosolic enzyme pyridoxine 5'-beta-D-glucoside hydrolase (PNG hydrolase) and by the brush border enzyme lactase phlorizin hydrolase (LPH) to release free pyridoxine (PN). This laboratory has previously shown that PNG hydrolase activity is inversely related to dietary vitamin B-6 in rats and guinea pigs. The current investigation was done to examine the effect of dietary PN on PNG hydrolytic activity and its distribution. Nutrient compositional differences between the AIN-76A and AIN-93G purified diets that were unrelated to vitamin B-6 were also examined in relation to PNG hydrolysis in rat small intestinal mucosa. Study one included rats (n = 29) that were fed the AIN-93G diet providing a range of PN concentrations for 5 wk. Rats (n = 49) in study two were fed either AIN-76A or AIN-93G each with graded concentrations of PN. In both studies, rat growth and plasma and liver pyridoxal 5'-phosphate (PLP) concentrations increased (P < 0.05) with increasing concentrations of dietary PN. PNG hydrolytic activity localized to the brush border membrane was five times that measured in the cytosol. Cytosolic PNG hydrolytic activity increased significantly with increasing dietary PN concentration in rats fed the AIN-76A, but not AIN-93G diet. Activity in the mucosal total membrane fraction did not increase in proportion to dietary PN concentration for either diet. Regardless of dietary PN concentration, the basal nutrient composition of the diets affected growth and PNG hydrolytic activity in intestinal mucosa. In contrast to previous results from this laboratory, intestinal hydrolytic activity toward PNG did not increase in vitamin B-6 deficient rats. PMID- 12730424 TI - Rye bread in the diet of pigs enhances the formation of enterolactone and increases its levels in plasma, urine and feces. AB - To obtain new insight into the quantitative and qualitative metabolism of rye and wheat lignans, we performed three series of experiments with catheterized pigs. Two diets with similar levels of dietary fiber and macronutrients but with contrasting levels of plant lignans (isolariciresinol, lariciresinol, matairesinol, pinoresinol, secoisolariciresinol and syringaresinol) were prepared from rye (high in lignans) and wheat (low in lignans) soft and crisp breads. In two series of experiments we quantified the uptake from the gut of enterolactone in four pigs fitted with catheters in the portal vein and mesenteric artery and with an ultrasonic flow probe attached to the portal vein to monitor the blood flow. In a third study with six pigs, we quantified the bioavailability of the plant lignans that can be converted to enterolactone (lariciresinol, matairesinol, pinoresinol, secoisolariciresinol and syringaresinol) and the concentration in the peripheral blood. Plant and mammalian lignans in diets and stool were analyzed by isotope dilution gas chromatography-mass spectrometry and enterolactone in plasma and urine determined by time-resolved fluoroimmunoassay. There was a significantly higher formation of enterolactone in pigs fed the rye diet, and higher fecal and urinary excretion and circulating levels of mammalian lignans than in pigs fed the wheat diet. The conversion of mammalian lignan precursors to enterolactone was 48% with the wheat diet and 60% with the rye diet. Mammalian lignans are absorbed by passive diffusion from the large intestine and a substantial fraction of the absorbed mammalian lignans undergoes enterohepatic circulation, resulting in low diurnal variation in plasma levels of enterolactone. PMID- 12730426 TI - The total branched-chain amino acid requirement in young healthy adult men determined by indicator amino acid oxidation by use of L-[1-13C]phenylalanine. AB - Previous recommendations for branched-chain amino acids (BCAA), based on nitrogen balance studies, were found to be low in a series of stable isotope-labeled amino acid studies. The BCAA requirement was increased in the new dietary reference intake (DRI) report on the basis of a series of stable isotope studies examining the requirement of leucine and valine individually, but not isoleucine. To reduce the possibility of interactions among these amino acids and imbalances in the mixture affecting the estimate of requirements, we decided to determine the requirement for the total BCAA of young healthy adult men, receiving a mixture of BCAA based on the proportion of these amino acids in egg protein, by use of indicator amino acid oxidation. Seven men were assigned to receive nine graded intakes of a BCAA mixture in random order: 34, 50, 66, 80, 100, 120, 140, 160 and 180 mg/(kg. d). The rate of release of (13)CO(2) from the oxidation of L-[1 (13)C]phenylalanine (F (13)CO(2)) was measured and a two-phase linear regression crossover model was applied to determine total BCAA requirement. The mean requirement and population-safe level (upper limit of 95% confidence interval) of the total BCAA were 144 and 210 mg/(kg. d), respectively. Based on the balance of BCAA in egg protein, our estimate for the mean leucine requirement is 55 mg/(kg. d), which is substantially higher than the 34 mg/(kg. d) recommended by the DRI. PMID- 12730425 TI - Early posthatch feeding stimulates satellite cell proliferation and skeletal muscle growth in turkey poults. AB - The effect of early posthatch feeding on skeletal muscle growth and satellite cell myogenesis was studied in turkey poults. Poults were either fed immediately posthatch or food-deprived for the first 48 h and then refed for the rest of the experiment. Body and breast muscle weights were lower in the starved poults than in fed controls throughout the experiment (P < 0.05). Cultures of breast muscle satellite cells revealed significantly higher DNA synthesis in the fed group than in the starved group as early as d 1 (P < 0.05). These levels continued to rise, reaching approximately 500-fold those of feed-deprived poults on d 4. In the latter group, thymidine incorporation peaked only on d 6, and then declined. Thereafter, it decreased to the same levels as those in the fed group. Satellite cell number per gram muscle increased until d 4, and was higher in the fed group than in the starved group (P < 0.05). Pax7 levels in cell cultures derived from the fed group were markedly higher than in the starved group on d 2 (P < 0.05). Myogenin levels in both culture and muscle were higher in the fed than in the starved groups until d 4 (P < 0.05). Phosphorylation of the survival factor Akt and cyclin-dependent kinase inhibitor p21 levels were higher in cells derived from the fed group relative to those from the starved group 48 h posthatch (P < 0.05). Similarly, Akt phosphorylation and insulin-like growth factor I (IGF-I) levels were significantly higher in the muscles of the fed group (P < 0.05). Together, these results suggest that immediate posthatch feeding of poults is critical for satellite cell survival and myogenesis probably via IGF-I. PMID- 12730427 TI - The methionine requirement is lower in neonatal piglets fed parenterally than in those fed enterally. AB - The requirements for the sulfur amino acids (SAA), methionine (Met) and cysteine (Cys), have seldom been determined in neonates and to our knowledge have not previously been determined directly in parenterally fed neonates. The sulfur amino acids are catabolized largely in the liver and kidney, and their metabolism by the gut has been studied less frequently. In the present research, the enteral and parenteral Met requirement was determined, without dietary Cys, using the indicator amino acid oxidation (IAAO) technique. Piglets [n = 32, 2 d old, 1.66 +/- 0.13 kg (SD)] received elemental diets containing adequate energy, phenylalanine (Phe) and excess tyrosine, with varied Met concentrations and no Cys. Diets were infused continuously via intravenous or intragastric catheters. Phenylalanine oxidation was determined during a primed, constant infusion of L-[1 (14)C]-Phe, by measuring expired (14)CO(2) and plasma specific radioactivity of Phe. For both parenteral and enteral groups, Phe oxidation (% of dose) decreased linearly (P < 0.01) as Met intake increased, then became low and unchanging. Using breakpoint analysis, the Met requirement was estimated to be 0.42 and 0.29 g/(kg. d) for enteral and parenteral feeding, respectively. Breakpoint analysis using absolute phenylalanine oxidation [ micro mol/(kg. h)] resulted in an estimation of the Met requirement of 0.44 and 0.26 g/(kg. d) for enteral and parenteral feeding, respectively. These data show that the parenteral Met requirement is approximately 69% of the enteral requirement and suggest that extraction of SAA by first-pass splanchnic metabolism may be responsible for this difference. PMID- 12730428 TI - Cod protein lowers the hepatic triglyceride secretion rate in rats. AB - The objective of the present study was to determine the combined effects of cod protein and fish oil on the modulation of triglyceride metabolism in rats, and to evaluate their potential mechanisms of action. Plasma and hepatic lipid concentrations, triglyceride (TG) secretion rates and postheparin plasma lipoprotein lipase (LPL) activity were determined in rats fed for 28 d diets varying in both protein (200 g/kg) and lipid (140 g/kg) sources: 1) casein menhaden oil, 2) casein-beef tallow, 3) cod protein-menhaden oil or 4) cod protein-beef tallow. Menhaden oil feeding diminished hepatic TG concentrations (P = 0.02), hepatic TG secretion rates (P = 0.003) and triglyceridemia (P = 0.02) compared with beef tallow. Hepatic TG concentrations (P = 0.05) and TG secretion rates (P = 0.04) were reduced in rats fed cod protein compared with those fed casein. The protein source did not exert an independent effect on triglyceridemia, whereas the combination of cod protein and menhaden oil resulted in 50% lower plasma TG compared with the casein-beef tallow mixture, whereas the combination of menhaden oil and casein did not significantly decrease triglyceridemia compared with casein-beef tallow. Menhaden oil (P = 0.005) and cod protein (P = 0.03) also lowered plasma cholesterol concentrations in comparison with beef tallow and casein, respectively. This was associated with a reduction in hepatic cholesterol concentrations when rats fed cod protein were compared with those fed casein (P = 0.006). No diet effect was observed on postheparin plasma LPL activity, but the activity of hepatic triglyceride lipase was reduced in rats fed menhaden oil compared with those fed beef tallow. These findings show that both cod protein and menhaden oil exert independent and beneficial effects on lipid metabolism in rats. PMID- 12730429 TI - Chronic consumption of a moderately low protein diet does not alter hematopoietic processes in young adult mice. AB - The current studies examined whether hematopoiesis in the bone marrow and T-cell development in the thymus were attenuated in young adult A/J mice fed a moderately low protein diet (MPD, 50 g protein/kg) for 15 wk compared with mice fed a control protein diet (CPD, 180 g protein/kg). Flow cytometric analyses using antibodies against CD31 and Ly-6C as well as CD4 and CD8 were performed to identify stem, mixed progenitor, erythroid, lymphoid, granuloid and monocytic compartments in the bone marrow and four thymocyte subsets, respectively. Chronic restriction of young adult mice to MPD neither decreased the cellularity nor altered the distribution of subpopulations in either primary tissue. Subsequently, a new set of mice were provided with CPD and a low protein diet (LPD, 25 g protein/kg). After 5 wk, body and thymus weights in LPD group were reduced 26 and 30%, respectively, which was accompanied by a 505% increase in serum corticosterone. Surprisingly, LPD did not alter the number or distribution of cells in the bone marrow and the percentages of thymocyte subsets, supporting the findings from the MPD group. We conclude that chronic consumption of a marginal protein diet by young adult mice does not disrupt hematopoietic processes. PMID- 12730430 TI - Dietary (n-3)/(n-6) fatty acid ratio: possible relationship to premenopausal but not postmenopausal breast cancer risk in U.S. women. AB - Recent research has suggested that an increased (n-3) fatty acid intake and/or increased (n-3)/(n-6) polyunsaturated fatty acid (PUFA) ratio in the diet is associated with a lower breast cancer risk. This case-control study investigated the association between intake of (n-3) and other fatty acids and the (n-3)/(n-6) PUFA ratio and breast cancer risk. After combining data from two related case control studies in Connecticut, we had information available on a total of 1119 women (565 cases and 554 controls). Cases were all histologically confirmed, incident breast carcinoma patients. Controls were hospital-based (Yale-New Haven Hospital study site) and population-based (Tolland County study site). Information on dietary intake was obtained through a validated food-frequency questionnaire. Standard multivariate methods were used to address the independent effects of specific fatty acids, fat classes and macronutrients on breast cancer risk. In the full study population, there were no significant trends for any macronutrient/fatty acid when comparing the highest to the lowest quartile of intake. When the analysis was restricted to premenopausal women, consumption of the highest compared with the lowest quartile of the (n-3)/(n-6) PUFA ratio was associated with a nonsignificant 41% lower risk of breast cancer [odds ratio (OR) = 0.59, 95% confidence interval (CI) 0.29, 1.19, P for trend = 0.09]. A higher (n 3)/(n-6) PUFA ratio was significantly associated with a lower risk of breast cancer when the data were restricted to the Tolland County (population-based) study site; OR = 0.50, 95% CI 0.27, 0.95, P for trend = 0.02. These results are consistent with the hypothesis that a higher (n-3)/(n-6) PUFA ratio may reduce the risk of breast cancer, especially in premenopausal women. PMID- 12730431 TI - Ethnicity modifies seasonal variations in birth weight and weight gain of infants. AB - Early growth is considered critical for the development of obesity and adulthood cardiovascular diseases. Because season and environmental temperature at birth have also been associated with obesity, it is important to examine whether birth weight and postnatal weight gain vary by season. Data from the National Collaborative Perinatal Project were used to assess the influence of season on birth weight and weight gain during the first 4 mo of life. The sample included 11,091 whites, 11,477 blacks, 1536 Puerto Ricans and 221 subjects of other ethnic groups born at full-term gestation. Black infants born in the fall had a significantly lower birth weight (3.12 +/- 0.42 kg) than those born in the winter (3.16 +/- 0.43 kg, P = 0.002). This difference was not found in the other ethnic groups. Additionally, weight gain (g/mo) for black and Puerto Rican infants during the first 4 mo of life was significantly lower for those born during the fall (black: 816 +/- 186; Puerto Rican: 820 +/- 181) compared to those born in the spring (black: 844 +/- 194, P < 0.001) and summer (Puerto Rican: 861 +/- 185, P < 0.04). Birth weight and early infancy weight gain varied by season and were modified by ethnicity. The potential importance of seasonal variations in pre- and postnatal growth was evaluated in this study. PMID- 12730432 TI - A longitudinal comparison of body composition by total body water and bioelectrical impedance in adolescent girls. AB - Previous studies have assessed the ability of bioelectrical impedance analysis (BIA) to estimate body composition cross-sectionally, but less is known about the ability of BIA to detect changes in body composition longitudinally over the adolescent growth period. Body composition was assessed by isotopic dilution of H(2)(18)O and BIA in 196 initially nonobese girls enrolled in a longitudinal study. Two prediction equations for use in our population of girls were developed, one for use premenarcheally and one for use postmenarcheally. We compared estimates from our equation with those derived from several published equations. Using longitudinal data analysis techniques, we estimated changes in fat-free mass (FFM) and percentage body fat (%BF) over time from BIA, compared with changes in FFM and % BF estimated by H(2)(18)O. A total of 422 measurements from 196 girls were available for analysis. Of the participants, 26% had one measurement of body composition, 43% had two measurements of body composition and 31% had three or more measurements of body composition. By either H(2)(18)O or BIA, the mean %BF at study entry was 23% (n = 196) and the mean %BF at 4 y postmenarche was 27% (n = 133). In our cohort, the best predictive equations to estimate FFM by BIA were: PREMENARCHE: FFM = -5.508 + (0.420 x height(2)/resistance) + (0.209 x weight) + (0.08593 x height) + (0.515 x black race) - (0.02273 x other race). POSTMENARCHE: FFM = -11.937 + (0.389 x height(2)/resistance) + (0.285 x weight) + (0.124 x height) + (0.543 x black race) + (0.393 x other race). Overall, we found that BIA provided accurate estimates of the change in both FFM and %BF over time. PMID- 12730433 TI - The role of passive transport in calcium absorption. PMID- 12730436 TI - Trace elements and nitric oxide function. AB - Nitric oxide (NO) has emerged over the last 15 y as a mammalian metabolic intermediate that is involved in the regulation of critical physiological functions such as blood vessel homeostasis, neuronal transmission and host response to infection. NO is synthesized by the enzyme nitric oxide synthase, which converts the amino acid L-arginine to citrulline and NO. NO functions in biological systems in two very important ways. First, it has been found to be a messenger by which cells communicate with one another (signal transduction), and second, it plays a critical role in the host response to infection. In this second function, it appears that the toxic properties of NO have been harnessed by the immune system to kill or at least slow the growth of invading organisms. The nonspecific chemical reactivity with key cellular targets is responsible for this action. In signaling, NO directly activates the enzyme soluble guanylate cyclase (sGC). Once activated, sGC converts GTP to cGMP and pyrophosphate. The cGMP formed is responsible for the well-documented actions of NO such as blood vessel dilation. With the initial discovery of NO signaling, several important questions emerged that centered largely on the issue of how a signaling system functions when the signaling agent is chemically reactive (short lived), highly diffusible and toxic. Critical, especially in signaling, are the control of NO biosynthesis and interaction with the biological receptors at a concentration that will not harm the host. Why did Nature choose NO for the roles it has? That question engenders only speculation. How does NO work (i.e., what does NO do, and how does it do it without harm yet with specificity)? Answers to these questions can now be offered as the molecular level details emerge to form an interesting picture. PMID- 12730437 TI - Scientific research: essential, but is it enough to combat world food insecurities? AB - Food and nutrition insecurity, which affects an estimated 815 million households (the majority in developing countries), is in large part due to micronutrient deficiencies. The magnitude of the problem, causes, consequences and cost effective solutions elucidated by scientists over the last few decades changed perceptions and drew political commitments in the 1990s to alleviate micronutrient deficiencies. Prevalence was reduced for iodine and vitamin A deficiency disorders largely through mandated universal fortification of salt with iodine and wide distribution of vitamin A supplements associated with immunization campaigns. Less progress was achieved in the control of iron deficiency. The challenge now is to move forward with interventions that are sustainable within the context of vulnerable communities, which possibly include applied biotechnology to enhance yields and micronutrient contents of staple foods. Scientific progress and political commitment are key factors, but consumer and public acceptance is key to sustainable progress. Achieving public confidence requires increased partnerships among scientists, policymakers, community leaders and consumers in the decision-making processes. PMID- 12730438 TI - Underwood Memorial Lecture: human zinc homeostasis: good but not perfect. AB - Three selected aspects of human zinc homeostasis and requirements are reviewed with special reference to studies undertaken by the author and his colleagues: 1) the implications for the calculation of physiologic requirements for zinc of the interrelationship between two key variables of zinc homeostasis, intestinal excretion of endogenous zinc and total absorbed zinc, are examined at levels of absorption below those necessary to meet physiologic requirements; 2) a method for deriving average dietary zinc requirements from zinc-stable isotope tracer/metabolic studies is illustrated with examples of studies being conducted in developing countries; and 3) the effect of reduction of high intakes of phytic acid on zinc bioavailability is examined with test meals prepared from low-phytic acid maize or isohybrid wild-type control maize. PMID- 12730439 TI - Trace elements and host defense: recent advances and continuing challenges. AB - Although it is widely recognized that essential trace elements are required for the differentiation, activation and performance of numerous functions of immune cells, the specific roles of these inorganic micronutrients in these processes remain largely undefined. New insights about the participation of zinc, iron and copper in the selection, maturation and early activation events of the immune cells have been gained by judicious use of available tools in analytical cell biology, molecular genetics and array technology. Also, randomly controlled clinical and community trials demonstrate that zinc supplementation can enhance immunocompetence and decrease the incidence and severity of some infections in individuals with diagnosed or suspected mild zinc deficiency. These exciting results provide an impetus to evaluate the potential benefits of supplementation programs for individuals and groups with suboptimal trace element status as a cost-effective means of reducing the risk of infectious diseases. PMID- 12730440 TI - Role of copper, zinc, selenium and tellurium in the cellular defense against oxidative and nitrosative stress. AB - The trace elements copper, zinc and selenium are linked together in cytosolic defense against reactive oxygen and nitrogen species. Copper, zinc-superoxide dismutase catalyzes the dismutation of superoxide to oxygen and hydrogen peroxide. The latter and other hydroperoxides are subsequently reduced by the selenoenzyme glutathione peroxidase (GPx). Cytosolic GPx can also act as a peroxynitrite reductase. The antioxidative functions of these trace elements are not confined to being constituents of enzymes: 1) copper and zinc ions may stimulate protective cellular stress-signaling pathways such as the antiapoptotic phosphoinositide-3-kinase/Akt cascade and may stabilize proteins, thereby rendering them less prone to oxidation; and 2) selenium does not only exist in the cell as selenocysteine (as in GPx) but also as selenomethionine, which is regularly present in low amounts in proteins in place of methionine. Selenomethionine catalyzes the reduction of peroxynitrite at the expense of glutathione. Also, low-molecular-weight organoselenium and organotellurium compounds of pharmacologic interest catalyze the reduction of hydroperoxides or peroxynitrite with various cellular reducing equivalents. PMID- 12730441 TI - Zinc-altered immune function. AB - Zinc is known to be essential for all highly proliferating cells in the human body, especially the immune system. A variety of in vivo and in vitro effects of zinc on immune cells mainly depend on the zinc concentration. All kinds of immune cells show decreased function after zinc depletion. In monocytes, all functions are impaired, whereas in natural killer cells, cytotoxicity is decreased, and in neutrophil granulocytes, phagocytosis is reduced. The normal functions of T cells are impaired, but autoreactivity and alloreactivity are increased. B cells undergo apoptosis. Impaired immune functions due to zinc deficiency are shown to be reversed by an adequate zinc supplementation, which must be adapted to the actual requirements of the patient. High dosages of zinc evoke negative effects on immune cells and show alterations that are similar to those observed with zinc deficiency. Furthermore, when peripheral blood mononuclear cells are incubated with zinc in vitro, the release of cytokines such as interleukins (IL)-1 and -6, tumor necrosis factor-alpha, soluble IL-2R and interferon-gamma is induced. In a concentration of 100 micro mol/L, zinc suppresses natural killer cell killing and T-cell functions whereas monocytes are activated directly, and in a concentration of 500 micro mol/L, zinc evokes a direct chemotactic activation of neutrophil granulocytes. All of these effects are discussed in this short overview. PMID- 12730442 TI - Selenium in the immune system. AB - Selenium as an essential component of selenocysteine-containing protein is involved in most aspects of cell biochemistry and function. As such, there is much potential for selenium to influence the immune system. For example, the antioxidant glutathione peroxidases are likely to protect neutrophils from oxygen derived radicals that are produced to kill ingested foreign organisms. When the functions of all selenoproteins are described, only then will it be possible to fully understand their role in maintaining optimal immune function. PMID- 12730443 TI - Cellular zinc and redox states converge in the metallothionein/thionein pair. AB - The paramount importance of zinc for a wide range of biological functions is based on its occurrence in thousands of known zinc proteins. To regulate the availability of zinc dynamically, eukaryotes have compartmentalized zinc and the metallothionein/thionein pair, which controls the pico- to nanomolar concentrations of metabolically active cellular zinc. Interactions of zinc with sulfur ligands of cysteines turn out to be critical both for tight binding and creation of a redox-active coordination environment from which the redox-inert zinc can be distributed. Biological oxidants such as disulfides and S nitrosothiols oxidize the zinc/thiolate clusters in metallothionein with concomitant zinc release. In addition, selenium compounds that have the capacity to form selenol(ate)s catalytically couple with the glutathione/glutathione disulfide and metallothionein/thionein redox pairs to either release or bind zinc. In this pathway, selenium expresses its antioxidant effects through redox catalysis in zinc metabolism. Selenium affects the redox state of thionein, an endogenous chelating agent. With its 20 cysteines, thionein contributes significantly to the zinc- and thiol-redox-buffering capacity of the cell. Thus, hitherto unknown interactions between the essential micronutrients zinc and selenium on the one hand and zinc and redox metabolism on the other are key features of the cellular homeostatic zinc system. PMID- 12730444 TI - Selenium deficiency and viral infection. AB - The discovery that the juvenile cardiomyopathy known as Keshan disease likely has a dual etiology that involves both a nutritional deficiency of the essential trace mineral selenium (Se) as well as an infection with an enterovirus provided the impetus for additional studies of relationships between nutrition and viral infection. An amyocarditic strain of coxsackievirus B3, CVB3/0, converted to virulence when it was inoculated into Se-deficient mice. This conversion was accompanied by changes in the genetic structure of the virus so that its genome closely resembled that of other known virulent CVB3 strains. Similar alterations in virulence and genomic composition of CVB3/0 could be observed in mice fed normal diets but genetically deprived of the antioxidant selenoenzyme glutathione peroxidase (knockout mice). More recent research has shown that a mild strain of influenza virus, influenza A/Bangkok/1/79, also exhibits increased virulence when given to Se-deficient mice. This increased virulence is accompanied by multiple changes in the viral genome in a segment previously thought to be relatively stable. Epidemic neuropathy in Cuba has features that suggest a combined nutritional/viral etiology. Further research, both basic and applied, is needed to assess properly the possible role of malnutrition in contributing to the emergence of novel viral diseases. PMID- 12730445 TI - Iron deficiency alters brain development and functioning. AB - Iron deficiency anemia in early life is related to altered behavioral and neural development. Studies in human infants suggest that this is an irreversible effect that may be related to changes in chemistry of neurotransmitters, organization and morphology of neuronal networks, and neurobiology of myelination. The acquisition of iron by the brain is an age-related and brain-region-dependent process with tightly controlled rates of movement of iron across the blood-brain barrier. Dopamine receptors and transporters are altered as are behaviors related to this neurotransmitter. The growing body of evidence suggests that brain iron deficiency in early life has multiple consequences in neurochemistry and neurobiology. PMID- 12730447 TI - Developmental consequences of trace mineral deficiencies in rodents: acute and long-term effects. AB - Approximately 3% of infants born have at least one serious congenital malformation. In the U.S., an average of 10 infants per thousand die before 1 y of life; about half of these deaths can be attributed to birth defects, low birth weight or prematurity. Although the causes of developmental abnormalities are clearly multifactorial in nature, we suggest that a common factor contributing to the occurrence of developmental abnormalities is suboptimal mineral nutrition during embryonic and fetal development. Using zinc and copper as examples, evidence is presented that nutritional deficiencies can rapidly affect the developing conceptus and result in gross structural abnormalities. Deficits of zinc or copper can result in rapid changes in cellular redox balance, tissue oxidative stress, inappropriate patterns of cell death, alterations in the migration of neural crest cells and changes in the expression of key patterning genes. In addition to well-recognized malformations, mineral deficiencies during perinatal development can result in behavioral, immunological and biochemical abnormalities that persist into adulthood. Although these persistent defects can in part be attributed to subtle morphological abnormalities, in other cases they may be secondary to epigenetic or developmental changes in DNA methylation patterns. Epigenetic defects combined with subtle morphological abnormalities can influence an individual's risk for certain chronic diseases and thus influence his or her risk for morbidity and mortality later in life. PMID- 12730446 TI - The evidence linking zinc deficiency with children's cognitive and motor functioning. AB - The role of zinc in children's cognitive and motor functioning is usually assessed by the response to supplementation in populations thought to be zinc deficient. A review of published zinc-supplementation trials that examined behavior and development identified one trial in fetuses, six trials in infants and toddlers and three trials in school-age children. The three studies that examined activity reported that zinc supplementation was associated with more activity. Of the five studies that examined motor development in infants and toddlers, one found improvements among very low-birth-weight infants, one found improvements in the quality of motor development and three found no impact. Of the four studies that examined mental development in infants and toddlers, three found no impact of zinc supplementation and one found that zinc-supplemented children had lower scores than control children. Among school-age children, one study found no impact of zinc supplementation on cognitive performance and two found a beneficial impact of neuropsychological processes, specifically reasoning. The evidence linking zinc deficiency to children's cognitive and motor functioning suggests a relationship among the most vulnerable children but lacks a clear consensus, highlighting the need for additional research into the timing of zinc deficiency and the co-occurrence with other micronutrient deficiencies. PMID- 12730448 TI - Molecular and cellular aspects of copper transport in developing mammals. AB - Copper is an essential trace element that requires tightly regulated homeostatic mechanisms to ensure adequate supplies without any toxic effects because of the ability of the metal ion to catalyze the formation of free radicals. The Cu ATPases, ATP7A and ATP7B, play an important role in the physiological regulation of copper. Adequate supplies of copper are particularly important in developing animals, and in humans this is illustrated by mutations of ATP7A that cause the copper deficiency condition Menkes disease, which is fatal in early childhood. In contrast, mutations in ATP7B result in the genetic toxicosis, Wilson disease. We propose that the physiological regulation of copper is accomplished mainly by the intracellular copper-regulated trafficking of the Cu-ATPases. This process allows the overall copper status in the body to be maintained when levels of copper in the diet alter. A study of the defects in mouse models of Menkes and Wilson diseases has demonstrated that both ATPases play an important role in supplying copper to the developing fetus and neonate. PMID- 12730449 TI - Zinc deficiency, infectious disease and mortality in the developing world. AB - Zinc deficiency places children in many low-income countries at increased risk of illness and death from infectious diseases. Randomized controlled trials of zinc supplementation provide the best estimate of this risk through demonstrated preventive benefits. In six of nine trials that evaluated prevention of diarrhea, significantly lower incidence of diarrhea occurred in the zinc group than in the controls; a pooled analysis demonstrated 18% (95% confidence interval, 7-28%) less diarrhea. In five trials, a lower rate of pneumonia infection was found in the zinc-supplemented groups, and there was some indication of a preventive effect in three trials with a clinical malaria outcome. Zinc was also found to have a therapeutic benefit in seven trials of acute diarrhea and five of persistent diarrhea. Studies to evaluate the effect of zinc supplementation on mortality are under way, but a recently published study from India identified a 68% reduction in mortality in small-for-gestational-age term infants that were supplemented with zinc from 1 to 9 mo of age. The important effects of zinc deficiency are now clear, and nutrition programs should address this prevalent problem. PMID- 12730450 TI - Genetically modified plants for improved trace element nutrition. AB - Deficiencies of iron and zinc are common worldwide. Various strategies have been used to combat these deficiencies including supplementation, food fortification and modification of food preparation and processing methods. A new possible strategy is to use biotechnology to improve trace element nutrition. Genetic engineering can be used in several ways; the most obvious is to increase the trace element content of staple foods such as cereals and legumes. This may be achieved by introduction of genes that code for trace element-binding proteins, overexpression of storage proteins already present and/or increased expression of proteins that are responsible for trace element uptake into plants. However, even very high levels of expression may not substantially increase the iron and zinc contents unless many atoms of trace elements are bound per protein molecule. Another possibility is to introduce a protein that specifically enhances trace element absorption even in the presence of naturally occurring inhibitors, thus improving bioavailability. Genetically modifying plants so that their contents of inhibitors of trace element absorption such as phytate are substantially reduced is another approach. Increasing the expression of compounds that enhance trace element absorption such as ascorbic acid is also a possibility, although this has received limited attention so far. Iron absorption may be increased by higher ascorbic or citric acid content but require overexpression of enzymes that are involved in the synthetic pathways. Finally, a combination of all of these approaches perhaps complemented with conventional breeding techniques may prove successful. PMID- 12730451 TI - Zinc supplementation and growth of the fetus and low birth weight infant. AB - Zinc deficiency limits growth in young children, and in animal models it also affects fetal growth. In animals, the effect of severe zinc deficiency on growth is evident. However, controlled trials of zinc supplementation during pregnancy in humans have not demonstrated consistent effects on weight at birth and/or duration of gestation. Those studies that have identified a positive effect of zinc on fetal growth were performed on low-socioeconomic or migrant groups in industrialized countries or in countries where diet and living conditions are improving. In contrast, in studies conducted either on populations with minimal risk of zinc deficiency or those suffering from multiple and severe nutritional deficiencies, no effects have been found. The potential confounding factors that may help explain the contradictory results include the following: age of pregnant women; presence of digestive diseases, other nutritional deficiencies, phytates or other factors that affect bioavailability; timing and duration of zinc supplementation and compliance with supplements. Few studies of zinc supplementation were conducted on infants that were born small for their gestational age or preterm; in this case, a positive effect on growth was shown. A study of zinc supplementation during pregnancy found reduced risk of diseases (diarrhea or impetigo) in small-for-gestational-age but not preterm infants. A second study demonstrated reduced mortality in small-for-gestational-age infants. We conclude that supplementation trials during human pregnancy do not provide conclusive evidence for a beneficial effect of zinc supplementation despite the experimental evidence that zinc deficiency may retard fetal growth or shorten pregnancy. However, early zinc supplementation in low birth weight or small-for gestational-age infants reveals an effective improvement in growth, which suggests a prenatal depletion or insufficient zinc intake to support catch-up growth postnatally. PMID- 12730452 TI - Exchangeable zinc pool size in infants is related to key variables of zinc homeostasis. AB - The exchangeable zinc pool (EZP) is the sum of the combined pools that exchange with zinc in the plasma within 48-72 h and is thought to be critical for zinc dependent biological processes. The size of the EZP in adults has been found to be positively related to dietary zinc intake, daily absorbed zinc and fecal excretion of endogenous zinc. In this study, we examine data on EZP size in relation to variables of zinc homeostasis in normal infants on different feeding regimens. Data from 45 male infants who participated in one of four different studies contribute to the analysis. The feeding regimens include exclusive breastfeeding (n = 9; age, 2-5 mo); breastfeeding plus modest supplementation with cow's milk-based formula (n = 16; age, 3-4 mo); exclusive formula feeding (n = 4; age, 3-4 mo) and exclusive breastfeeding plus complementary foods (n =16; age, 7 mo). Fractional absorption was determined by fecal monitoring after oral administration of zinc-stable isotopes. Urine enrichment 4-8 d posttracer was used to determine endogenous fecal zinc (7-mo-old infants excepted) and EZP size. Univariate correlations and multivariate regression analyses were performed between EZP and age, weight, dietary zinc intake, fractional absorption, total absorbed zinc and endogenous fecal zinc. Results include no significant relationship between EZP size and age, body weight or fractional absorption but a positive relationship with daily absorbed zinc and endogenous fecal zinc excretion. We conclude that the amount of absorbed zinc is not tightly regulated, and that endogenous fecal zinc is dependent on and responsive to the zinc status of the organism. PMID- 12730453 TI - Trace element uptake and distribution in plants. AB - There are similarities between mammals and plants in the absorption and transport of trace elements. The chemistry of trace element uptake from food sources in both cases is based on the thermodynamics of adsorption on charged solid surfaces embedded in a solution phase of charged ions and metal-binding ligands together with redox systems in the case of iron and some other elements. Constitutive absorption systems function in nutrient uptake during normal conditions, and inducible "turbo" systems increase the supply of a particular nutrient during deficiency. Iron uptake is the most studied of the micronutrients, and divides the plant kingdom into two groups: dicotyledonous plants have a turbo system that is an upregulated version of the constitutive system, which consists of a membrane-bound reductase and an ATP-driven hydrogen ion extrusion pump; and monocotyledonous plants have a constitutive system similar to that of the dicots, but with an inducible system remarkably different that uses the mugeneic acid class of phytosiderophores (PS). The PS system may in fact be an important port of entry for iron from an iron-rich but exceedingly iron-insoluble lithosphere into the iron-starved biosphere. Absorption of trace metals in these graminaceous plants is normally via divalent ion channels after reduction in the plasma membrane. Once absorbed, iron can be stored in plants as phytoferritin or transported to active sites by transport-specific ligands. The transport of iron and zinc into seeds is dominated by the phloem sap system, which has a high pH that requires chelation of heavy metals. Loading into grains involves three or four genes each that control chelation, membrane transport and deposition as phytate. PMID- 12730454 TI - Trace mineral bioavailability in ruminants. AB - Absorption of selenium and copper is much lower in ruminants than in nonruminants. The low absorption of these minerals in ruminants is due to modifications that occur in the rumen environment. Selenium bioavailability is reduced by high dietary sulfur and the presence of cyanogenetic glycosides in certain legumes. Feeding organic selenium from selenomethionine or selenized yeast results in much higher tissue and milk selenium concentrations than are obtained with selenite. High dietary molybdenum in combination with moderate to high dietary sulfur results in formation of thiomolybdates in the rumen. Thiomolybdates greatly reduce copper absorption, and certain thiomolybdate species can be absorbed and interfere systemically with copper metabolism. Independent of molybdenum, high dietary sulfur reduces copper absorption perhaps via formation of copper sulfide. High dietary iron also reduces copper bioavailability. Dietary factors that affect bioavailability of zinc in ruminants are not well defined. Phytate does not affect zinc absorption in ruminants because microbial phytase in the rumen degrades phytate. Manganese is very poorly absorbed in ruminants, and limited research suggests that high dietary calcium and phosphorus may reduce manganese absorption. PMID- 12730455 TI - Novel roles for iron regulatory proteins in the adaptive response to iron deficiency. AB - Iron regulatory proteins (IRP) modulate the use of mRNA-encoding proteins that are involved in the transport, storage and use of iron. Several new potential mRNA targets for IRP were recently identified: divalent metal transporter-1 (DMT 1) and ferroportin, which are critical regulators of iron absorption in the gut and of iron cycling between various tissues of the body. Although this may extend the reach of IRP to other processes that are important for maintaining body iron homeostasis, the extent to which IRP modulate other physiological processes that are altered in response to changes in iron availability is not clear. However, in the past several years, targets for IRP and IRP-like proteins were identified in eukaryotes and prokaryotes in the tricarboxylic acid (TCA) cycle and electron transport chain. In mammals, this includes the mRNA that encodes the TCA-cycle enzyme mitochondrial aconitase (m-acon). Recent work established that m-acon expression is translationally regulated by iron in a manner that is strongly correlated with IRP RNA-binding activity. Interestingly, these studies also demonstrate that IRP regulate their mRNA targets in a hierarchical manner. The changes in m-acon synthesis and abundance in liver during iron deficiency fail to affect TCA-cycle capacity but are associated with a significant upregulation of mitochondrial export of radiolabeled citrate. We conclude that IRP are required for the regulation of physiological pathways that include but are not limited to iron metabolism, and as such, IRP are critical factors in the adaptive response to iron deficiency. PMID- 12730457 TI - Regulation of zinc metabolism and genomic outcomes. AB - Differential mRNA display and cDNA array analysis have identified zinc-regulated genes in small intestine, thymus and monocytes. The vast majority of the transcriptome is not influenced by dietary zinc intake, high or low. Of the genes that are zinc regulated, most are involved in signal transduction (particularly influencing the immune response), responses to stress and redox, growth and energy utilization. Among the genes identified are uroguanylin (UG), cholecystokinin, lymphocyte-specific protein tyrosine kinase (LCK), T-cell cytokine receptor, heat shock proteins and the DNA damage repair and recombination protein-23B. Zinc transporters (ZnT) help regulate the supply of this micronutrient to maintain cellular functions. Expression of ZnT-1 and -2 is regulated by dietary zinc in many organs including small intestine and kidney. ZnT-4 is ubiquitously expressed but is refractory to zinc intake. Expression of ZnT-1, -2 and -4 changes markedly during gestation and lactation from highly abundant to undetectable. Each ZnT has an endosomal-like appearance in the tissues examined. Upregulation of ZnT-1 and ZnT-2 by dietary zinc strongly implicates these transporters in zinc acquisition and/or storage for subsequent systemic needs. THP-1 cells were used as a model to examine the response of human cells to changes in zinc status. Based on mRNA quantities, Zip1 and ZnT-5 were the most highly expressed. Zinc depletion of these cells decreased expression of all transporters except Zip2, where expression increased markedly. Collectively, these findings provide a genomic footprint upon which to address the biological and clinical significance of zinc and new avenues for status assessment. PMID- 12730456 TI - Selenoprotein metabolism and function: evidence for more than one function for selenoprotein P. AB - Biological functions of selenium are exerted by selenoproteins that contain selenocysteine in their primary structure. Selenocysteine is synthesized and inserted into proteins cotranslationally by a complex process. Families of selenoproteins include the glutathione peroxidases, the iodothyronine deiodinases and the thioredoxin reductases. These are redox enzymes that take advantage of the chemical properties of selenium to catalyze, respectively, removal of hydroperoxides by glutathione, deiodination of thyroid hormones and support of cellular processes requiring reduction of disulfides. Approximately 10 additional selenoproteins have been identified. One of them, selenoprotein P, is an extracellular protein that contains most of the selenium in plasma. It associates with endothelial cells, probably through its heparin-binding properties. Selenoprotein P has been postulated to protect against oxidative injury and to transport selenium from the liver to peripheral tissues. Selenium-dependent protection against diquat-induced liver necrosis and lipid peroxidation in the rat correlates with the presence of selenoprotein P. Recent results support a transport function. When (75)SeO(3)(2-) was administered intravenously to rats, liver tissue took up (75)Se within minutes, associated with a rapid decline in plasma (75)Se. Brain tissue did not begin accumulating (75)Se until (75)Se labeled selenoprotein P had begun appearing in the plasma after 30 min. These results suggest that tissues like liver can take up small-molecule forms of selenium whereas presence of the element in selenoprotein P facilitates uptake by tissues like brain. Thus, there is evidence for both antioxidant and selenium transport functions of selenoprotein P. PMID- 12730458 TI - Genetic defects in copper metabolism. AB - Genetic defects in copper metabolism highlight the delicate balance mammalian systems have developed to maintain normal copper homeostasis. Menkes disease, the mottled mouse, the Atox-1-deficient mouse and the ctr1 knockout mouse reveal the importance of adequate copper intake during embryogenesis and early development, especially in the central nervous system. The toxicity associated with excess copper as manifest in Wilson disease, the toxic milk mouse, the LEC rat and copper toxicosis in the Bedlington terrier demonstrate the profound cellular susceptibility to copper overload, in particular, in the brain and liver. Ceruloplasmin (Cp) contains 95% of the copper found in human serum, and inherited loss of this protein results in diabetes, retinal degeneration and neurodegeneration. Despite normal copper metabolism, aceruloplasminemic patients and the Cp knockout mouse have disturbed iron homeostasis and mild hepatic copper retention. These genetic disorders of copper metabolism provide valuable insight into the mechanisms regulating copper homeostasis and models to further dissect the role of this essential metal in health and disease. PMID- 12730459 TI - Multiple regulatory mechanisms maintain zinc homeostasis in Saccharomyces cerevisiae. AB - Zinc is an essential nutrient, yet it is toxic if it accumulates in excess amounts within cells; therefore the intracellular labile zinc content of cells is tightly controlled. In Saccharomyces cerevisiae, zinc homeostasis is regulated by the controlled activity of zinc uptake transporters in the plasma membrane and transporters responsible for intracellular zinc compartmentalization. The activity of these transporters is regulated at both transcriptional and posttranscriptional levels in response to zinc. These different mechanisms work together to precisely balance zinc uptake and its storage and utilization. PMID- 12730460 TI - Health effects and risk assessment of arsenic. AB - Humans can be exposed to arsenic (As) through the intake of air, food and water. Although food is usually the major source of As exposure for people, most adverse effects are seen after As exposure from drinking water. The two main reasons for this situation are that most food arsenicals are organic and have little or no toxicity, and in many cases, As exposures from drinking water sources are to the more toxic inorganic form and occur at relatively high doses, e.g., hundreds of micrograms per day. In various parts of the world, As in drinking water is associated with such effects as gastroenteritis, neurological manifestations, vascular changes, diabetes and cancers (bladder, lung, liver, kidney and prostate). After reviewing the As database, the U.S. Environmental Protection Agency promulgated a maximum contaminant level for As in drinking water of 10 micro g/L. PMID- 12730461 TI - Nutritional factors may modify the toxic action of methyl mercury in fish-eating populations. AB - The historical record of clinical cases of methyl mercury poisoning dates back to the 19th century when the first chemical synthesis occurred. The potent fungicidal properties of both methyl and the closely related ethyl mercury compound were subsequently discovered, which led to widespread agricultural application for prevention of fungal infection in seed grain. Several catastrophic outbreaks of poisoning occurred in the mid-20th century when the treated seed grain was mistakenly used to prepare homemade bread. The largest outbreak took place in rural Iraq in the early 1970s. Human poisonings also occurred in Japan due to the release of methyl mercury into bodies of fresh and ocean water. The most infamous outbreak occurred in the area of Minamata Bay: methyl mercury, which was unwittingly discharged into the ocean water, avidly accumulated in the aquatic food chain to such an extent that people who consumed fish were severely poisoned. Today, human exposure to methyl mercury occurs from consumption of fish and sea mammals. Inorganic mercury that is present in aquatic sediments is methylated by microorganisms and accumulates in the aquatic food chain. Although no cases of clinical poisoning have been reported, a number of epidemiological studies have been carried out that raise the possibility of prenatal damage. Previous studies (especially the Iraq outbreak) indicate that the prenatal stage of the life cycle is the most vulnerable. However, ongoing epidemiological studies of heavy fish consumers of the Seychelles Islands in the Indian Ocean do not reveal adverse effects. To the contrary, the results of some developmental tests that were conducted on prenatally exposed children indicate beneficial outcomes that correlate with mercury levels during pregnancy. This article discusses the potential role of micronutrients in fish as a plausible explanation for these findings. PMID- 12730462 TI - The metabolic tune-up: metabolic harmony and disease prevention. AB - An optimum intake of micronutrients and metabolites, which varies with age and genetic constitution, would tune up metabolism and give a marked increase in health, particularly for the poor and elderly, at little cost. 1) DNA damage. Inadequate intake of folic acid causes millions of uracils to be incorporated into the DNA of each cell with associated chromosome breaks, essentially producing a radiation mimic. Deficiencies of the metabolically connected vitamins B-6 and B-12, which are also widespread, also cause uracil incorporation and chromosome breaks. Inadequate iron intake (2 billion women in the world; 25% of U.S. menstruating women) causes oxidants to leak from mitochondria and damages mitochondria and mitochondrial DNA. Inadequate zinc intake (approximately 10% in the U.S.) causes oxidation and DNA damage in human cells. 2) The K(m) concept. Approximately 50 different human genetic diseases that are due to a poorer binding affinity (K(m)) of the mutant enzyme for its coenzyme can be remedied by feeding high-dose B vitamins, which raise levels of the corresponding coenzyme. Many polymorphisms also result in a lowered affinity of enzyme for coenzyme. 3) Mitochondrial oxidative decay with age. This decay, which is a major contributor to aging, can be ameliorated by feeding old rats the normal mitochondrial metabolites acetyl carnitine and lipoic acid at high levels. They restore the K(m) for acetyl carnitine transferase and the velocity of the reaction as well as mitochondrial function; reduce levels of oxidants, neuron RNA oxidation and mutagenic aldehydes; and increase old-rat ambulatory activity and cognition. PMID- 12730463 TI - Ferritin: at the crossroads of iron and oxygen metabolism. AB - Iron and oxygen are central to terrestrial life. Aqueous iron and oxygen chemistry will produce a ferric ion trillions of times less soluble than cell iron concentrations, along with radical forms of oxygen that are toxic. In the physiological environment, many proteins have evolved to transport iron or modulate the redox chemistry of iron that transforms oxygen in useful biochemical reactions. Only one protein, ferritin, evolved to concentrate iron to levels needed in aerobic metabolism. Reversible formation and dissolution of a solid nanomineral-hydrated, iron oxide is the main function of ferritin, which additionally detoxifies excess iron and possibly dioxygen and reactive oxygen. Ferritin is a large multifunctional, multisubunit protein with eight Fe transport pores, 12 mineral nucleation sites and up to 24 oxidase sites that produce mineral precursors from ferrous iron and oxygen. Regulation of ferritin synthesis in animals uses both DNA and mRNA controls and genes encoding two types of related subunits with: 1) catalytically active (H) or 2) inactive (L) oxidase sites. Ferritin with varying H/L ratios is related to cell-specific iron and oxygen homeostasis. H-ferritin oxidase activity accelerates rates of iron mineralization in ferritins and, in animals, ferritin produces H(2)O(2) as a byproduct. Properties of ferritin mRNA and ferritin protein pore structure are new targets for manipulating iron homeostasis. Recent observations of the high bioavailability of iron in soybean ferritin and efficient utilization of soybean and ferritin iron by iron-deficient animals, and of soybean iron by humans with borderline deficiency, indicate a role for ferritin in managing global iron deficiency in humans. PMID- 12730465 TI - Marginal dietary selenium intakes in the UK: are there functional consequences? AB - Much data indicate that overt selenium deficiency induces a number of pathologies in animals and humans. The effects of chronic marginal undernutrition of this element are unclear, although it has been argued that such subjects will be at increased risk of developing various cancers. The dietary intake of selenium in the UK has fallen over the last 25 years, although no functional consequences of this have been recognized. Recent data demonstrate that restoration of selenium intakes in UK subjects induces biochemical effects with increased activity of selenium-dependent enzymes. Whether such biochemical changes are associated with functional changes is currently unclear and the subject of current investigation. PMID- 12730464 TI - Iron and copper interactions in development and the effect on pregnancy outcome. AB - During pregnancy, nutrients are transferred from mother to fetus across the placenta. The mechanisms whereby this occurs, and the adaptations that occur in response to deficiency or overload of iron (Fe) and copper (Cu) are examined in this review. Fe deficiency during pregnancy is common and has serious consequences both in the short and the long term such as fetal growth retardation and cardiovascular problems in the adult offspring. Similarly, Cu deficiency, although not so common, also has deleterious effects. The placenta minimizes the effect of the deficiency by up-regulating the proteins involved in Fe transfer. For example, transferrin receptor levels increase inversely to maternal Fe levels. Divalent metal transporter 1 (DMT1) mRNA in the iron-responsive element (IRE) regulated, but not the non-IRE regulated form is increased, as is the placenta Cu oxidase. Conversely, iron-regulated gene 1 (IREG1) expression is not affected. Fe deficiency increases Cu levels in maternal liver, serum and placenta, but has much less effect in the fetal serum and liver. Apart from maternal ceruloplasmin, mRNA levels of Cu-related proteins are not changed. The Cu oxidase, which we suggest fulfils the function of hephaestin in placenta, is regulated by Cu as well as by Fe. Fe deficiency also has marked effects on cytokine levels in the placenta. Tumor necrosis factor alpha (TNFalpha) and TNFalpha receptor 1 (TNFalphaR1) levels both increase. The data show that altering Fe status has a marked effect on metabolism of other metals and of other important mediators of cell function. This is particularly important during pregnancy, when the developing fetus is very vulnerable to inappropriate micronutrient status. PMID- 12730466 TI - Approaches to improve iron bioavailability from complementary foods. AB - The importance of trace element bioavailability in the etiology of nutritional deficiencies, for example in the etiology of iron deficiency and iron deficiency anemia, can be expected to be most pronounced in individuals with high requirements. Of special concern is the situation in poor communities where infants and young children are consuming monotonous, cereal-based diets. Traditionally, cereal-based gruels are often one of the first semisolid foods to be introduced into the infant's diet. These foods can be expected to have low energy and nutrient density as well as low bioavailability of iron due to the presence of phytic acid. Ascorbic acid is a potent enhancer of non-heme iron absorption that can overcome the inhibiting effect of phytic acid when present in high enough quantities. However, home prepared complementary foods based on cereals and legumes contain negligible amounts of ascorbic acid unless ascorbic acid-rich foods are mixed with the cereal or consumed at the same time. Different approaches to improve iron bioavailability from plant-based complementary foods, e.g., by enzymatic degradation of phytic acid and/or by increased consumption of ascorbic acid-rich foods, should be explored and adapted to local conditions. In addition, there is a need to evaluate efficacy and effectiveness of strategies to increase the dietary intake of bioavailable iron by dietary diversification and food fortification under realistic conditions. PMID- 12730467 TI - Assessment of the trace element status of individuals and populations: the example of zinc and copper. AB - This paper describes the proceedings of a workshop that was convened at the 11th International Symposium on Trace Elements in Man and Animals (TEMA-11) symposium to review recent advances concerning the assessment of the trace element status of individuals and populations, using zinc and copper as the primary examples to illustrate basic principles and recent advances in assessment methods. The workshop was initiated with a brief review of the importance of zinc nutriture for human health and a discussion of the likely common occurrence of zinc deficiency worldwide. This overview was followed by presentations on selected issues concerning the assessment of zinc status, with particular attention devoted to dietary assessment techniques, the use of isotopic tracers to assess zinc homeostasis and the relationship of these methods to biochemical indicators of zinc status. Because relatively little information is available on zinc toxicity, the discussion concerning the definition of excess intake of trace elements focused primarily on recent work concerning risk assessment of copper toxicity. PMID- 12730468 TI - Design and quality control issues related to dietary assessment, randomized clinical trials and meta-analysis of field-based studies in developing countries. AB - The essential design and quality control aspects of dietary assessment, randomized controlled trials (RCT) in developing countries and their subsequent meta-analysis are reviewed. Dietary assessment protocols consist of three stages: measurement of food intakes using a method appropriate for the study objectives, calculation of nutrient intakes and finally an evaluation of their nutrient adequacy. The latter may involve: 1) assessment of dietary diversity (average number of different foods consumed per day) and selected indices of dietary quality such as intakes of animal source foods (g/d); 2) percentage of energy from protein, fat and animal protein; 3) selected nutrient densities; and 4) dietary phytate:zinc molar ratio, as well as the prevalence of inadequate nutrient intakes calculated using a probability approach. To establish causality between the nutrient inadequacies identified and adverse health outcomes, RCT must be undertaken. A prerequisite of RCT is double-blind randomization, a procedure designed to eliminate biases arising from baseline confounding variables, unintended interventions and ascertainment bias. Results from existing RCT can be summarized via meta-analysis to gain a better understanding of the population relationship. Meta-analysis is a statistical technique involving linear models or generalized linear models, which can be performed after locating the individual studies, and selecting and abstracting all the necessary data. PMID- 12730469 TI - Analytical methods: improvements, advancements and new horizons. AB - The workshop and exhibits dealing with analytical methods were selected to highlight the current state of the art in elemental analysis. The presentations in the first part of the workshop described approaches and advances important to the analysis of trace minerals. These presentations included: 1) two approaches to mass spectrometry, inductively coupled plasma and accelerator mass spectrometry; 2) use of nuclear magnetic resonance in studies of mineral function; and 3) the use and limitations of fluorescent probes in studies of metal uptake and regulation. In the second part of the workshop, the International Atomic Energy's contributions to nutritional "metrology" were described. Advances in instrumentation over the past decade have led to extraordinary improvements in the precision and sensitivity of mineral analyses. The ability to address isotopic speciation at such low levels sets the stage for numerous novel approaches in the assessment of trace mineral function. PMID- 12730470 TI - Integrating trace element metabolism from the cell to the whole organism. AB - The redox chemistry of copper (Cu) makes this both a powerful enzyme catalyst and a dangerous reactant that generates hydroxyl radical. Although virtually all cells from microbes to mammals must acquire Cu to drive important biochemical reactions, the potential toxicity of Cu demands an exquisite level of vectorial transport and homeostatic control. Our laboratory is interested in how organisms acquire Cu through the action of high-affinity plasma membrane Cu transporters of the copper transport protein (Ctr) class of proteins. We have isolated Ctr Cu transporters from baker's yeast and fission yeast and from flies, mice and mammals. This review will focus on understanding how the Ctr high-affinity Cu transport proteins function, from their biochemical mechanism of action in yeast and cultured metazoan cells to their roles in Cu delivery and mammalian embryonic development. PMID- 12730471 TI - Trace element biology: the knowledge base and its application for the nutrition of individuals and populations. AB - Impressive strides are being made in the understanding of trace element metabolism and function. This is underscored by the many contributions in these proceedings. However, not so impressive are: i) the precise recognition of mild trace element deficiencies and how to establish their functional consequences, possibly confounded by concurrent trace element inadequacies, are difficult to assess, ii) approaches to the quantitative determination of requirements for trace elements remain unsatisfactory and archaic, in so many ways, iii) our understanding of the extent of the biological basis for the variation in requirements among apparently similar individuals is poor, and iv) much needs to be learned about the quantitative extent to which genetic, epigenetic and dietary factors interact to determine the nutritional phenotype. Some ideas are presented as to how we might embrace, in the context of a reconstructive approach, the exciting new knowledge and related techniques emerging during the postgenome era and develop new paradigms for assessing trace element needs and status, and for establishing effective nutrient intake under different conditions of complex genotype-environment interactions. Metabolites are functional cellular entities and I also urge a vigorous application of metabolomics and of metabolic profiling that is closely linked with genomics, proteomics, trace element kinetics and system analysis, as components of the new integrative paradigm. We need to understand the system and its strategy, not only the molecular details of its component parts and its individual controls. An interdisciplinary research and teaching enterprise will be necessary to best achieve this aim. All of this is related to our common goal to promote, through expanded biological knowledge and its effective application, the enhanced role of trace elements for human well being. PMID- 12730472 TI - Nutrition as a preventative strategy against adverse pregnancy outcomes. Introduction. PMID- 12730473 TI - The epidemiology of adverse pregnancy outcomes: an overview. AB - This paper provides an overview of the occurrence, etiology and temporal trends of adverse pregnancy outcomes. Disparities between developed and developing countries are highlighted for maternal mortality, infant mortality, stillbirth and low birth weight. The higher rate of low birth weight in developing countries is primarily due to intrauterine growth restriction rather than preterm birth. Much of the excess intrauterine growth restriction is caused by short maternal stature, low prepregnancy body mass index and low gestational weight gain (due to low energy intake). No important contribution has been established for micronutrient intake, nor have different fetal growth trajectories been demonstrated to reflect the timing of exposure to nutritional or other etiologic factors. Infant mortality has declined substantially over time both in developed and developing countries despite no decline (and even an increase) in low birth weight. Several developed countries have reported a temporal increase in fetal growth in infants born at term, a reduction in stillbirth rates and prevention of neural tube defects. More progress is required, however, in understanding the etiology and prevention of preterm birth. PMID- 12730474 TI - The plausibility of micronutrient deficiencies being a significant contributing factor to the occurrence of pregnancy complications. AB - Numerous studies support the concept that a major cause of pregnancy complications can be suboptimal embryonic and fetal nutrition. Although the negative effects of diets low in energy on pregnancy outcome are well documented, less clear are the effects of diets that are low in one or more essential micronutrients. However, several observational and intervention studies suggest that diets low in essential vitamins and minerals can pose a significant reproductive risk in diverse human populations. Although maternal nutritional deficiencies typically occur as a result of low dietary intakes of essential nutrients, nutritional deficiencies at the level of the conceptus can arise through multiple mechanisms. Evidence from experimental animals supports the concept that in addition to primary deficiencies, secondary embryonic and fetal nutritional deficiencies can be caused by diverse factors including genetics, maternal disease, toxicant insults and physiological stressors that can trigger a maternal acute phase response. These secondary responses may be significant contributors to the occurrence of birth defects. An implication of the above is that the frequency and severity of pregnancy complications may be reduced through an improvement in the micronutrient status of the mother. PMID- 12730476 TI - Nutritional interventions during pregnancy for the prevention or treatment of impaired fetal growth: an overview of randomized controlled trials. AB - This paper reviews the efficacy of nutrition interventions to prevent or treat impaired fetal growth. Searches were made for Cochrane systematic reviews and randomized controlled trials published before October 2002. Balanced protein energy supplementation reduced the risk of small for gestational age (SGA) by 30% (95%CI: 20% to 43%) while one trial conducted in New York, U.S., reported a negative effect of high protein supplementation on SGA (RR 1.58; 95%CI: 1.03 2.41). Calcium supplementation protected against low birth weight (RR 0.83; 95%CI: 0.71-0.98). Micronutrient supplements did not affect birth weight, except for magnesium supplementation, which reduced the risk of SGA by 30%. This finding, however, needs or be interpreted with caution because of methodological issues in the data analysis. Programmatic recommendations can be made only for intervening with balanced protein energy supplements, especially in population with a high prevalence of undernutrition. Research is needed to determine the efficacy of multiple micronutrient supplementation and the effect of single micronutrients supplementation on specific growth outcomes such as fetal organ and bone growth. In addition, the public health relevance of these outcomes and their relation to morbidity need to be evaluated. PMID- 12730475 TI - Nutritional interventions during pregnancy for the prevention or treatment of maternal morbidity and preterm delivery: an overview of randomized controlled trials. AB - This overview assesses the effectiveness of nutritional interventions to prevent or treat maternal morbidity, mortality and preterm delivery. Cochrane systematic reviews and other up-to-date systematic reviews and individual randomized controlled trials were sought. Searches were carried out up to July 2002. Iron and folate supplements reduce anemia and should be included in antenatal care programs. Calcium supplementation to women at high risk of hypertension during pregnancy or low calcium intake reduced the incidence of both preeclampsia and hypertension. Fish oil and vitamins E and C are promising for preventing preeclampsia and preterm delivery and need further testing. Vitamin A and beta carotene reduced maternal mortality in a large trial; ongoing trials should provide further evaluation. No specific nutrient supplementation was identified for reducing preterm delivery. Nutritional advice, magnesium, fish oil and zinc supplementation appear promising and should be tested alone or together in methodologically sound randomized controlled trials. Anema in pregnancy can be prevented and treated effectively. Considering the multifactorial etiology of the other conditions evaluated, it is unlikely that any specific nutrient on its own, blanket interventions or magic bullets will prevent or treat preeclampsia, hemorrhage, obstructed labor, infections, preterm delivery or death during pregnancy. The few promising interventions for specific outcomes should be tested or reconsidered when results of ongoing trials become available. Until then, women and their families should receive support to improve their diets as a general health rule, which is a basic human right. PMID- 12730477 TI - Characteristics of randomized controlled trials included in systematic reviews of nutritional interventions reporting maternal morbidity, mortality, preterm delivery, intrauterine growth restriction and small for gestational age and birth weight outcomes. PMID- 12730478 TI - Potential cost-effectiveness of nutrition interventions to prevent adverse pregnancy outcomes in the developing world. AB - The potential cost-effectiveness of antenatal nutrition interventions to improve pregnancy outcomes in the developing world has not undergone formal evaluation. Furthermore, the effectiveness of antenatal care in improving maternal or fetal and neonatal health has been questioned. However, reasonably compelling evidence from randomized trials shows that nutrition interventions can prevent both infant (iodine supplementation) and maternal (vitamin A and beta-carotene supplementation) deaths, and informal analysis suggests that the cost effectiveness of nutrition interventions would be comparable and, in some cases, markedly superior to several standard antenatal interventions. Future efforts to establish the cost-effectiveness of nutrition interventions in developing countries will depend on conducting large, pragmatic clinical trials that use region- and resource-appropriate interventions with mortality or valid, incontrovertibly severe morbidity endpoints. If such trials establish effectiveness, credible cost-effectiveness analyses can then be performed. PMID- 12730479 TI - The plausibility of micronutrient deficiency in relationship to perinatal infection. AB - Infection has a major effect on adverse pregnancy outcomes, and this relationship appears strongest among populations that suffer from malnutrition. The most likely mediating factor linking this association is the effect of nutritional status on various host defense mechanisms. These mechanisms include intact skin and epithelial surfaces, phagocytosis by macrophages and neutrophils, cell mediated protection by T cells and natural killer cells and antibody production by B cells. Deficiencies of virtually every vitamin and mineral and protein energy malnutrition have been shown to negatively affect some or several host defense functions. There is therefore no question that a plausible relationship exists between micronutrient deficiency and infection-mediated adverse pregnancy outcomes. However, proof of this relationship is generally not available. PMID- 12730480 TI - Assessing micronutrient status in the presence of inflammation. AB - Measurement of micronutrient status in the presence of inflammation is difficult for several reasons. Changes in levels of acute phase proteins are associated with increased plasma levels of some indicators of micronutrient status, such as ferritin, and decrease of others, such as retinol. Alterations in the plasma levels of acute phase proteins can occur from hemodilution, sequestration and increased or decreased rates of synthesis and breakdown. How much these relate to functional deficiency is not known. Assays that are less perturbed by inflammation, such as the transferrin receptor assay, and adjustment of plasma micronutrient levels according to different cutoff levels for acute phase proteins are helpful but they do not enable precise assessment of micronutrient status among individuals who are infected. Improving assessment of micronutrient status is important if micronutrient interventions are to be targeted to those with the greatest need. PMID- 12730481 TI - Infection-related morbidities in the mother, fetus and neonate. AB - Only partially understood host defense mechanisms operate against infections affecting maternal and fetal morbidity. Subclinical ascending infections through the lower female genital tract are predominant worldwide. Important micronutrient deficiencies may prevail in low-income countries where these infections are much more common than in high-income countries. Important morbidities related to poor perinatal outcome both for the mother and for the fetus and newborn comprise preterm birth, prelabor rupture of membranes, placental abruption (predelivery detachment of the placenta), postpartum sepsis and maternal anemia. In the fetus, sepsis and intrauterine growth retardation are suspected to be consequences of ascending maternal infections. In the newborn, septicemia and respiratory disorders as well as some neurological disorders seem to be consequences of such ascending genital infections in the pregnant woman. It is concluded that much more attention should be given to efforts to elucidate the host defense mechanisms and antimicrobial barriers from the vagina through the cervix, fetal membranes and amniotic fluid including the early fetal immunocompetence in the second and the third trimester of pregnancy. PMID- 12730482 TI - Pregnancy, nutrition and parasitic diseases. AB - In the developing world, young women, pregnant women, and their infants and children frequently experience a cycle where undernutrition (macronutrient and micronutrient) and repeated infection, including parasitic infections, lead to adverse consequences that can continue from one generation to the next. Among parasitic infections, malaria and intestinal helminths coexist widely with micronutrient deficiencies and contribute importantly to anemia and this cycle of retarded growth and development. In somewhat more limited or focal geographic settings, other parasitic diseases (e.g., schistosomiasis, filariasis) contribute similarly to this cycle. It is undoubtedly much better to enter a pregnancy free of infection and nutritionally replete than the various alternatives. Existing intervention strategies for micronutrient support and for the control of common parasitic infections before or during pregnancy, particularly malaria and intestinal helminths, should be followed. However, further research to identify barriers and priority approaches to achieving this goal remain very important in resource-poor settings where targeted public health efforts are required. PMID- 12730483 TI - Micronutrients and intrauterine infection, preterm birth and the fetal inflammatory response syndrome. AB - Prematurity is the leading cause of perinatal morbidity and mortality worldwide. Intrauterine infection has emerged as a major cause of premature labor and delivery. It has been estimated that 25% of all preterm deliveries occur to mothers who have microbial invasion of the amniotic cavity, although these infections are mostly subclinical in nature. This article describes the pathways leading to intrauterine infection, microbiology, frequency and clinical consequences of infection. The pathophysiology of the fetal inflammatory response syndrome is reviewed, as is its relationship to long-term handicap, such as cerebral palsy and bronchopulmonary dysplasia. A possible role for two micronutrients, vitamins C and E, in the prevention of the preterm prelabor rupture of membranes and the consequences of fetal inflammation is considered. Research needs are listed. PMID- 12730484 TI - Gestational diabetes and insulin resistance: role in short- and long-term implications for mother and fetus. AB - Gestational diabetes and obesity are the common metabolic abnormalities occurring during pregnancy. Decreased maternal pregravid insulin sensitivity (insulin resistance) coupled with an inadequate insulin response are the pathophysiological mechanisms underlying the development of gestational diabetes. Insulin-regulated carbohydrate, lipid and protein metabolism are all affected to a variable degree. Decreased maternal insulin sensitivity in women with gestational diabetes may increase nutrient availability to the fetus, possibly accounting for an increased risk of fetal overgrowth and adiposity. Epidemiological studies from Europe show an increased risk of the insulin resistance syndrome in adults who were low birth weight at delivery. However, in the United States over the past 20 y there has been a significant 33% increase in the incidence of type 2 diabetes, which has been associated with a parallel increase in obesity. All age groups have been affected but the most dramatic increases have occurred in adolescents. The relationship between decreased maternal insulin sensitivity and fetal overgrowth particularly in obese women and women with gestational diabetes may help explain the increased incidence of adolescent obesity and related glucose intolerance in the offspring of these women. In this review, we address 1) the pathophysiology of gestational diabetes, 2) the changes in maternal insulin sensitivity during pregnancy that effect maternal accretion of adipose tissue and energy expenditure, 3) the influence of maternal metabolic environment on fetal growth, 4) the life-long effect of being born at either extreme of the birth weight continuum and 5) micronutrients and decreased insulin sensitivity during pregnancy. PMID- 12730485 TI - Nutrient involvement in preeclampsia. AB - Preeclampsia is a pregnancy-specific condition that increases maternal and infant mortality and morbidity. It is diagnosed by new-onset increased blood pressure and proteinuria during gestation; for many years these markers were the sole targets for study. More recently, increased attention to the multisystemic nature of the syndrome with involvement of almost all organs, activation of coagulation and increased sensitivity to pressor agents has expanded understanding of the disorder. The epidemiology of preeclampsia, being more common in poor women, long ago suggested that nutrients might be involved in the disorder. Numerous conflicting hypotheses were advanced but the testing of these hypotheses has either been done poorly or not at all. Review of the available data indicates very few studies that provide useful insights. In many studies the syndrome is poorly defined and in most studies nutritional data (questionnaires or biomarkers) are obtained on women with the clinical syndrome. In overtly preeclamptic women it is impossible to decipher cause from effect. Nonetheless, current concepts of the genesis of preeclampsia that include endothelial dysfunction, inflammatory activation, oxidative stress and predisposing maternal factors provide targets for well-designed nutritional investigation. In this review the current concepts of the pathogenesis of preeclampsia are reviewed and available data are assessed in light of these concepts. Targets for nutritional investigation based on the current knowledge of pathophysiology are suggested. PMID- 12730486 TI - Micronutrients and the bone mineral content of the mother, fetus and newborn. AB - The fluxes of the primary bone-forming minerals, calcium, phosphorus, magnesium and zinc, across the placenta and through breast milk place considerable demands on maternal mineral economy. Increases in food consumption, elevated gastrointestinal absorption, decreased mineral excretion and mobilization of tissue stores are several possible biological strategies for meeting these extra mineral requirements. This paper presents a review of the evidence on the extent to which these strategies apply in the human situation, the mechanisms by which they occur, the limitations imposed by maternal diet and vitamin D status and the possible consequences for the growth of the infant and bone health of the mother. On the strength of current evidence it appears that pregnancy and lactation are associated with physiological adaptive changes in mineral metabolism that are independent of maternal mineral supply within the range of normal dietary intakes. These processes provide the minerals necessary for fetal growth and breast milk production without requiring an increase in maternal dietary intake or compromising maternal bone health in the long term. This may not apply to pregnant women whose mineral intakes or sunlight exposure are marginal. As a vehicle for promoting optimal growth and bone mineral content of infants, supplementation of lactating women with minerals or vitamin D is unlikely to prove effective. The situation in pregnancy is less certain. Until more studies have been conducted, a precautionary case can be made for targeted supplementation of pregnant women who have very low intakes of calcium or who are at risk of vitamin D deficiency. PMID- 12730488 TI - Role of psychosocial and nutritional stress on poor pregnancy outcome. AB - Epidemiological evidence suggests that maternal psychosocial stress, strenuous physical activity and fasting are independent risk factors for preterm birth and low birth weight. Data from clinical studies consistently demonstrate that women in preterm labor have significantly elevated levels of corticotropin-releasing hormone compared with age-matched control subjects. Because production of corticotropin-releasing hormone appears to be stress sensitive, this neuropeptide may play a critical role in the physiological mediation among stressful experiences, work stress and fasting and risk of preterm birth. In addition to the direct effect of elevated corticotropin-releasing hormone on the initiation of labor, it may have an immunomodulatory effect such that women with high levels of corticotropin-releasing hormone may be more susceptible to infection or the pathological consequences of infection. We review the epidemiological data linking maternal stress, physical stain and fasting to preterm birth and low birth weight and review the plausible biological pathways through which these exposures may increase risk of preterm birth. The timing of these exposures is considered important. Future research and clinical programs addressing these exposures must consider assessments and interventions before pregnancy. PMID- 12730487 TI - Iron and oxidative stress in pregnancy. AB - Pregnancy, mostly because of the mitochondria-rich placenta, is a condition that favors oxidative stress. Transitional metals, especially iron, which is particularly abundant in the placenta, are important in the production of free radicals. Protective mechanisms against free radical generation and damage increase throughout pregnancy and protect the fetus, which, however, is subjected to a degree of oxidative stress. Oxidative stress peaks by the second trimester of pregnancy, ending what appears to be a vulnerable period for fetal health and gestational progress. Conditions restricted to pregnancy, such as gestational hypertension, insulin resistance and diabetes, exhibit exaggerated indications of free radical damage. Antioxidants as well as avoidance of iron excess ameliorate maternal and early fetal damage. In rats both iron deficiency and excess result in free radical mitochondrial damage. Estimates of gestational iron requirements and of the proportion of iron absorbed from different iron supplemental doses suggest that with present supplementation schemes the intestinal mucosal cells are constantly exposed to unabsorbed iron excess and oxidative stress. Unpublished work carried out in Mexico City with nonanemic women at midpregnancy indicates that 60 mg/d of iron increases the risk of hemoconcentration, low birth weight and premature birth and produces a progressive decline in plasma copper. These risks are not observed in women supplemented with 120 mg iron once or twice per week. Studies on the influence of iron supplementation schemes on oxidative stress are needed. PMID- 12730489 TI - Strenuous work, nutrition and adverse pregnancy outcomes: a brief review. AB - This brief review explores the available epidemiologic data to investigate the question of whether strenuous work by women during pregnancy in developing countries influences micronutrient status and thereby increase risks of adverse pregnancy outcomes. Some data exist on the potential relationship between strenuous work or physical activity and nutrient compromise, strenuous work or physical activity and adverse reproductive outcomes and micronutrient intakes or status and adverse reproductive outcomes. No substantial body of data exists that has directly investigated the potential causal path of whether strenuous work during pregnancy alters micronutrient status leading to adverse reproductive outcome. Search of the literature identified only a few papers from developing countries that provided even remotely related data on the topic. Thus, the available data are insufficient for drawing firm inferences that strenuous work, in a developing country, alters a pregnant woman's nutritional status and therefore affects her risk of an adverse pregnancy outcome. Effects on nutritional status, micronutrients in particular, of pregnant women from strenuous physical activities at work or in other lifestyle events require further study in developing countries. PMID- 12730490 TI - Cigarette smoking, alcohol use and adverse pregnancy outcomes: implications for micronutrient supplementation. AB - This literature review examines whether smoking or alcohol use during pregnancy increases maternal micronutrient requirements and whether smoking or alcohol use interacts with micronutrient deficiencies to affect pregnancy outcomes. Studies suggest that vitamin C requirements increase for pregnant smokers. Studies also indicate that beta-carotene, vitamin B-12, vitamin B-6 and folate concentrations appear lower in pregnant smokers than in pregnant nonsmokers, although it is unclear whether lower serum concentrations are due to increased requirements, lower dietary or supplement intakes or other factors. Experimental animal studies suggest that iron supplementation partially ameliorates impaired fetal growth caused by cadmium, a heavy metal inhaled from cigarette smoke, but studies in humans have not substantiated cadmium's effect on fetal growth. Animal studies also suggest chronic alcohol consumption at levels of 20-50% of energy intake during pregnancy may mobilize fetal vitamin A concentration from the liver and result in increases in vitamin A in fetal organs and subsequent defects. Evidence is lacking, however, on whether zinc metabolism is altered by alcohol intake during pregnancy. Health care practitioners should consider increasing nutrient levels in pregnant women who do not meet the Recommended Dietary Allowances through their diet. Future studies that examine the nutrient levels of women exposed to cigarette smoke and alcohol should control for dietary intake. In addition, randomized controlled studies of the health impact of micronutrient supplementation in pregnant women should consider stratification by exposure to cigarette smoke and alcohol use. PMID- 12730492 TI - Some thoughts on body mass index, micronutrient intakes and pregnancy outcome. AB - A low prepregnancy body mass index is one of strongest predictors of adverse pregnancy outcomes such as preterm birth and fetal growth retardation. A low body mass interacts with other risk factors such as smoking and stress to increase risk of these outcomes, whereas zinc supplementation and low-dose aspirin increase birth weight in thin but not normal-size women. The association between maternal thinness and adverse pregnancy outcomes may be mediated more by a low plasma volume than by decreased protein or energy status. Maternal micronutrient status may partially mediate plasma volume expansion in pregnancy. Therefore, improving maternal micronutrient status may reduce adverse outcomes through this mechanism. PMID- 12730491 TI - The risk of maternal nutritional depletion and poor outcomes increases in early or closely spaced pregnancies. AB - An adequate supply of nutrients is probably the single most important environmental factor affecting pregnancy outcome. Women with early or closely spaced pregnancies are at increased risk of entering a reproductive cycle with reduced reserves. Maternal nutrient depletion may contribute to the increased incidence of preterm births and fetal growth retardation among these women as well as the increased risk of maternal mortality and morbidity. In the past, it was assumed that the fetus functioned as a parasite and withdrew its nutritional needs from maternal tissues. Studies in both animals and humans demonstrate, however, that if the maternal nutrient supply is inadequate, the delicate balance between maternal and fetal needs is disturbed and a state of biological competition exists. Furthermore, maternal nutritional status at conception influences how nutrients are partitioned between the mother and fetal dyad. In severe deficiencies maternal nutrition is given preference; in a marginal state the fetal compartment is favored. Although the studies of nutrient partitioning have focused on energy and protein, the partitioning of micronutrients may also be influenced by the maternal nutritional status. Marginal intakes of iron and folic acid during the reproductive period induce a poor maternal status for these nutrients during the interpregnancy interval. Poor iron and folic acid status has also been linked to preterm births and fetal growth retardation. Supplementation with food and micronutrients during the interpregnancy period may improve pregnancy outcomes and maternal health among women with early or closely spaced pregnancies. PMID- 12730493 TI - Regulation of fetal growth by the somatotrophic axis. AB - Suboptimal fetal growth is associated with higher fetal mortality and with higher neonatal morbidity and mortality. It increases the likelihood of premature birth that in turn further compounds perinatal health risks. Moreover, an abnormal fetal environment, as reflected in an altered birth size phenotype, increases the propensity for disease in childhood and adulthood. Fetal growth represents the culmination of interaction between the fetal genome and the in utero environment determined by maternal-placental function. The role of endocrine and metabolic factors in mediating this interaction will be reviewed. There is also evidence that fetal growth, as measured in late gestation, is dependent not only on the maternal environment but on events that occurred during the periconceptual period. Thus, fetal growth not only reflects the immediate fetal environment but events surrounding conception and embryonic life. PMID- 12730494 TI - Micronutrients and fetal growth. AB - Fetal undernutrition affects large numbers of infants in developing countries, with adverse consequences for their immediate survival and lifelong health. It manifests as intrauterine growth retardation (IUGR), defined as birth weight <10th percentile, which probably underestimates the number failing to achieve full growth potential. Birth weight is a crude measure of the dynamic process of fetal growth and does not capture effects of fetal undernutrition on body composition and the development of specific tissues. The link between maternal nutrition and fetal nutrition is indirect. The fetus is nourished by a complex supply line that includes the mother's diet and absorption, endocrine status and metabolism, cardiovascular adaptations to pregnancy and placental function. Micronutrients are essential for growth, and maternal micronutrient deficiency, frequently multiple in developing countries, may be an important cause of IUGR. Supplementation of undernourished mothers with micronutrients has several benefits but there is little hard evidence of improved fetal growth. However, this has been inadequately tested. Most trials have only used single micronutrients and many were inconclusive because of methodological problems. Several food-based studies (some uncontrolled) suggest benefits from improving maternal dietary quality with micronutrient-dense foods. One trial of a multivitamin supplement (HIV-positive mothers, Tanzania) showed increased birth weight and fewer fetal deaths. Well-conducted randomized controlled trials of adequate sample size and including measures of effectiveness are needed in populations at high risk of micronutrient deficiency and IUGR and should include food-based interventions and better measurements of fetal growth, maternal metabolism, and long-term outcomes in the offspring. PMID- 12730495 TI - Micronutrient status during pregnancy and outcomes for newborn infants in developing countries. AB - More than 9 million neonatal deaths occur each year, 98% of them in developing countries. Neonatal deaths account for two-thirds of deaths in infancy and 40% of deaths before age 5 y. The major direct causes of neonatal death are infections, preterm delivery and asphyxia. Important indirect causes include low birth weight and hypothermia. The present body of work on multiple micronutrient interventions is not sufficient for us to draw conclusions on their effects on neonatal well being. Because studies have generally concentrated on single micronutrients and a range of outcomes, this paper reviews the findings for individual nutrients and then summarizes the situation. The evidence for the contribution of micronutrient deficiencies to perinatal mortality and duration of gestation is limited, and the evidence base for individual micronutrient effects on neonatal mortality and morbidity is patchy. To translate knowledge into policy, community evaluations of effect and an expanded evidence base that includes affordability, acceptability and scalability are also required. A balance between supply-side and demand-side interventions must be struck, with an emphasis on effect and sustainability. Among the key requirements are randomized, controlled community effectiveness trials of the effect of micronutrient supplementation in pregnancy on perinatal mortality and neurodevelopment, studies on improving adherence and studies on the relation between micronutrient deficiencies and sepsis and neonatal encephalopathy. It would also be helpful to look at mechanisms for bringing the periconceptional period within the ambit of trials. PMID- 12730497 TI - Influence of satellite data uncertainties on the detection of externally forced climate change. AB - Two independent analyses of the same satellite-based radiative emissions data yield tropospheric temperature trends that differ by 0.1 degrees C per decade over 1979 to 2001. The troposphere warms appreciably in one satellite data set, while the other data set shows little overall change. These satellite data uncertainties are important in studies seeking to identify human effects on climate. A model-predicted "fingerprint" of combined anthropogenic and natural effects is statistically detectable only in the satellite data set with a warming troposphere. Our findings show that claimed inconsistencies between model predictions and satellite tropospheric temperature data (and between the latter and surface data) may be an artifact of data uncertainties. PMID- 12730498 TI - Derivation of oocytes from mouse embryonic stem cells. AB - Continuation of mammalian species requires the formation and development of the sexually dimorphic germ cells. Cultured embryonic stem cells are generally considered pluripotent rather than totipotent because of the failure to detect germline cells under differentiating conditions. Here we show that mouse embryonic stem cells in culture can develop into oogonia that enter meiosis, recruit adjacent cells to form follicle-like structures, and later develop into blastocysts. Oogenesis in culture should contribute to various areas, including nuclear transfer and manipulation of the germ line, and advance studies on fertility treatment and germ and somatic cell interaction and differentiation. PMID- 12730499 TI - Recruitment of HIV and its receptors to dendritic cell-T cell junctions. AB - Monocyte-derived dendritic cells (MDDCs) can efficiently bind and transfer HIV infectivity without themselves becoming infected. Using live-cell microscopy, we found that HIV was recruited to sites of cell contact in MDDCs. Analysis of conjugates between MDDCs and T cells revealed that, in the absence of antigen specific signaling, the HIV receptors CD4, CCR5, and CXCR4 on the T cell were recruited to the interface while the MDDCs concentrated HIV to the same region. We propose that contact between dendritic cells and T cells facilitates transmission of HIV by locally concentrating virus, receptor, and coreceptor during the formation of an infectious synapse. PMID- 12730502 TI - Cytokine balance in kidney tissue from lupus nephritis patients. AB - OBJECTIVE: To identify the balance of Th1/Th2 cytokine expression in the kidney and evaluate the difference in cytokine balance between patients with lupus nephritis WHO classes IV and V. METHODS: The expression of the CD40 molecule on cultured human mesangial cells was assessed by flow cytometry after stimulation with interferon gamma (IFN-gamma) or other cytokines. Frozen sections of kidney tissue from 10 patients with lupus nephritis and two non-SLE patients (with minimal-change disease) were stained with monoclonal antibodies for interleukin (IL)-4, IL-10, IL-12, IFN-gamma, CD4, CD8, CD40, CD68 and CD40L. RESULTS: CD40 expression of cultured mesangial cells was up-regulated by IFN-gamma, but was not down-regulated in the presence of the Th2 cytokines IL-4 and IL-10. In the glomeruli, CD40 expression and the ratios of IFN-gamma-/IL-10-, IL-12-/IL-4- and (IFN-gamma+IL-12)/(IL-4+IL-10)-positive cells were significantly higher in class IV than in class V lupus nephritis (P < 0.05). Also CD40, IFN-gamma and the activity index derived from the renal biopsy were closely correlated. CONCLUSION: IFN-gamma may contribute to the pathogenesis of proliferative glomerulonephritis by the up-regulation of CD40 and the activation of the cellular immune response in human lupus. PMID- 12730501 TI - The Genome sequence of the SARS-associated coronavirus. AB - We sequenced the 29,751-base genome of the severe acute respiratory syndrome (SARS)-associated coronavirus known as the Tor2 isolate. The genome sequence reveals that this coronavirus is only moderately related to other known coronaviruses, including two human coronaviruses, HCoV-OC43 and HCoV-229E. Phylogenetic analysis of the predicted viral proteins indicates that the virus does not closely resemble any of the three previously known groups of coronaviruses. The genome sequence will aid in the diagnosis of SARS virus infection in humans and potential animal hosts (using polymerase chain reaction and immunological tests), in the development of antivirals (including neutralizing antibodies), and in the identification of putative epitopes for vaccine development. PMID- 12730500 TI - Characterization of a novel coronavirus associated with severe acute respiratory syndrome. AB - In March 2003, a novel coronavirus (SARS-CoV) was discovered in association with cases of severe acute respiratory syndrome (SARS). The sequence of the complete genome of SARS-CoV was determined, and the initial characterization of the viral genome is presented in this report. The genome of SARS-CoV is 29,727 nucleotides in length and has 11 open reading frames, and its genome organization is similar to that of other coronaviruses. Phylogenetic analyses and sequence comparisons showed that SARS-CoV is not closely related to any of the previously characterized coronaviruses. PMID- 12730503 TI - Rheumatoid factor is the strongest predictor of radiological progression of rheumatoid arthritis in a three-year prospective study in community-recruited patients. AB - OBJECTIVE: To evaluate the predictive value of clinical, biological and radiological parameters for the prognosis of rheumatoid arthritis (RA) in a community-recruited cohort. METHODS: Ninety-one patients (mean age 49 yr, female/male ratio 2.9) with RA of limited duration (median 2 yr), 80% recruited from the community, were prospectively enrolled in 1996 (T1) and followed until 1999 (T2). Data collected at T1 were demographic characteristics, Ritchie articular index (RAI), extra-articular manifestations, Health Assessment Questionnaire (HAQ) score, C-reactive protein (CRP) and autoantibodies (autoAbs) [rheumatoid factors (RF), detected by latex fixation test and ELISA (IgM, IgA and IgG isotypes), anti-filaggrin, detected by immunofluorescence (anti-keratin antibodies, AKA; anti-perinuclear factor antibodies, APF) and ELISA (anti citrullinated rat filaggrin antibodies, ACRFA), anti-Sa, anti-calpastatin recognizing the 27 C-terminal fragment (ACAST-C27) and domain I (ACAST-DI), anti cardiolipin (ACL), antineutrophil cytoplasmic antibodies (ANCA), anti-annexin V (aANX V) and anti-Ro]. Hands were radiographed at T1 and T2, and read using the Sharp method as modified by van der Heijde. The main assessment criterion was progression of radiologically detected damage between T1 and T2. RESULTS: At T1, RA activity was mild (RAI 11/78; mean CRP 14 mg/ml), with minor functional disability (HAQ 0.8/3) and mild X-ray destruction (mean total Sharp score 9.2/280). At T1, 96% of the patients were on treatment (prednisone 72%, DMARDs 95%). The latex test detected autoAb in 46% of patients, RF-IgM was detected in 51%, RF-IgA in 36%, RF-IgG in 32%, AKA in 33%, APF in 45%, ACRFA in 45%, ACAST C27 in 14%, ACAST-DI in 5%, anti-Sa in 22%, ACL in 3%, ANCA in 28%, aANX V in 9% and anti-Ro in 2%. At T2, the mean total Sharp score was 22.9. According to univariate analysis, T1 parameters associated with the independent variable were RAI, HAQ, CRP, latex test positivity and T1 Sharp scores. Multivariate analysis retained only latex test positivity and, to a lesser degree, joint-space narrowing score as independent predictors of radiological progression. CONCLUSION: RF is the main factor that can predict radiological progression in community cases of RA of limited duration. PMID- 12730505 TI - Musculoskeletal assessment of general medical in-patients--joints still crying out for attention. AB - OBJECTIVES: The aim of this study was to assess the extent of musculoskeletal assessment (history and examination) amongst medical in-patients and to determine the effects that GALS (gait, arms, legs, spine) teaching has had on this. METHODS: General medical in-patients were interviewed and examined. Case notes were reviewed to determine diagnoses and assess documentation of clinical signs and/or symptoms. Doctors were interviewed to determine whether they had been taught the GALS locomotor screen, assess their confidence in examining the musculoskeletal system and explore their attitudes to musculoskeletal problems. RESULTS: The presence or absence of locomotor symptoms was recorded in 50% of the 100 patients, whilst signs were recorded in 20%; 63% of all the patients had locomotor symptoms and/or signs. Relevant musculoskeletal history was missed in 49% of the patients, whilst signs were missed in 78%; 42% of those with musculoskeletal conditions would have benefited from additional treatment. Teaching of the GALS screen significantly increased doctors' confidence in examination of the locomotor system. CONCLUSIONS: Active musculoskeletal problems are seen frequently amongst medical in-patients. There is a significant discrepancy between the number of patients with clinical symptoms and signs and the frequency with which they are detected and treated. PMID- 12730504 TI - Prevalence of Helicobacter pylori in NSAID users with gastric ulcer. AB - OBJECTIVE: Regarding the interaction of Helicobacter pylori and non-steroidal anti-inflammatory drugs (NSAIDs), we cannot accept unanimous conclusions in inducing gastric ulcer. We therefore evaluated the role of Helicobacter pylori and NSAIDs in inducing gastric ulcer. METHODS: Dyspeptic patients receiving NSAIDs underwent endoscopic examination. Gastric ulcer formation and H. pylori status were investigated. Biopsy specimens from the antrum and lower body of the stomach were prepared for the rapid urease test and pathological evaluation. Anti H. pylori antibody was measured by enzyme-linked immunosorbent assay. RESULTS: Two hundred and twenty-six patients receiving NSAIDs (220 chronic and six on demand users) underwent gastrofibrescopic examination. There were 110 patients with gastric ulcer and 111 non-ulcer patients with gastritis. The remaining five patients had neither. NSAID users with gastric ulcer showed a low prevalence of H. pylori compared with those without them [55/110 (50.0%) vs 79/111 (71.2%), P < 0.01]. The same tendency was seen when patients receiving low-dose aspirin and those with rheumatoid arthritis were analysed separately [13/29 (44.8%) vs 50/62 (80.6%), P < 0.01, and 11/33 (33.3%) vs 16/26 (61.5%), P < 0.06 with Yates' correction, respectively]. CONCLUSION: Helicobacter pylori infection appeared to be a risk factor for developing gastritis, but we found no evidence that it increases gastric ulcer formation in NSAID users with dyspepsia. PMID- 12730506 TI - Exploring the reciprocal relationship between immunity and inflammation in chronic inflammatory arthritis. AB - Experimental models seeking to explore how susceptible individuals develop rheumatoid arthritis (RA) propose that genetic and environmental factors shape a complex series of molecular and cellular interactions leading to a chronic inflammatory response. T lymphocytes and MHC class II genes have featured prominently in these models. More recent studies have suggested that perpetuation of inflammation in a disease-susceptible host might occur through failure to down regulate the inflammatory process. One prediction from this model is that effective mechanisms of immunoregulation might be most easily investigated in non susceptible individuals. However, this has been difficult to study in man. Based on the observation that extended MHC haplotypes are strongly associated with RA in different ethnic groups, I have explored the function of human MHC-encoded genes in transgenic mice using two different experimental approaches. First, by comparing the molecular interactions between disease-associated or non-associated HLA-DR4 molecules and CD4+ T lymphocytes, it has been possible to gain insight into how immune responses in non-susceptible individuals might differ from T-cell responses observed in a susceptible host. This has been achieved using transgenic mice expressing RA disease-associated and non-associated human HLA class II molecules. Secondly, the effects of prolonged exposure of T cells to the proinflammatory cytokine tumour necrosis factor alpha (TNF) have been studied in vitro and in vivo, focusing on T-cell receptor (TCR) signalling and effector responses. In studies of HLA class II transgenic mice, the major differences between disease-associated and non-associated alleles in terms of T-cell responses occur at the level of presentation of antigenic peptides, and the sustained expression of inflammatory cytokines such as TNF. Chronic exposure of T cells to inflammatory cytokines such as TNF induces a phenotype which resembles RA synovial T cells, including the induction of non-deletional and reversible hyporesponsiveness to TCR ligation and uncoupling of proximal TCR signal transduction pathways. The experimental findings are consistent with a model in which HLA class II-driven inflammatory cytokine expression uncouples TCR signalling pathways in the susceptible host in such a way as to profoundly suppress proliferative and immunoregulatory cytokine responses, while at the same time promoting cell survival and effector responses. PMID- 12730507 TI - Osteoarthritis pain and weather. AB - OBJECTIVE: To evaluate the association between weather (barometric pressure, precipitation and temperature) and pain among individuals with osteoarthritis (OA) (n=154) at the following sites: neck, hand, shoulder, knee and foot. METHODS: This prospective study evaluated men and women, aged 49-90 yr, participating in a community-based, osteoarthritis exercise study (June 1998 January 2002). Weekly self-reported pain scores were collected using a visual analogue scale. Statistical tests, including regression and correlation analyses, were conducted. P values < 0.001 were considered significant. RESULTS: The total number of pain recordings varied by site, ranging from 2269 (feet) to 6061 (hands). The mean temperature was 23 degrees C with a low of 0 degrees C and a high of 36 degrees C. Precipitation levels ranged from 0.00-21.08 cm, with a mean of 0.36 cm. Most associations explored produced non-significant findings. However, among women with hand OA, higher pain was significantly associated with days of rising barometric pressure (P < 0.001). CONCLUSION: Among a population of exercisers aged 49 yr and older, overall these findings did not support the hypothesis that weather is associated with pain. While some associations were suggestive of a relationship, largely these findings indicate that weather is quite modestly, if at all, associated with pain from OA. PMID- 12730508 TI - Risk factors for new-onset low back pain amongst cohorts of newly employed workers. AB - OBJECTIVES: To test the hypothesis that work-related mechanical, psychosocial and physical environment factors would predict new-onset low back pain (LBP) in newly employed workers. METHODS: A total of 1186 newly employed workers were recruited from a variety of occupational settings. Those who were free from LBP at baseline were identified. Subjects were followed up at 12 and 24 months. Work-related mechanical, psychosocial and physical environment exposures were measured. Generalized estimating equations were used to assess predictors of new-onset LBP. RESULTS: New-onset LBP was reported by 119 (19%) and 81 (19%) subjects at 12 and 24 months, respectively. Several work-related mechanical exposures predicted new onset LBP including lifting heavy weights with one or two hands, lifting heavy weights at or above shoulder level, pulling heavy weights, kneeling or squatting for 15 min or longer. Of the psychosocial factors examined, stressful and monotonous work significantly predicted symptom onset. In addition, hot working conditions and pain at other sites also predicted new-onset LBP. On multivariate analysis these risks were only moderately attenuated but the 95% confidence intervals excluded unity only for the latter, non-mechanical, exposures. CONCLUSION: In this cohort of newly employed workers, from a range of occupations, several aspects of the work-place environment, other than mechanical factors, were important in predicting new-onset LBP. These results emphasize that interventions aimed at reducing the occurrence of LBP are likely to be most successful if they intervene across these domains. PMID- 12730509 TI - Association of polymorphisms of the tumour necrosis factor receptors I and II and rheumatoid arthritis. AB - OBJECTIVE: To assess the role of polymorphisms of the tumour necrosis factor (TNF) receptors, TNF-RI (p55) and TNF-RII (p75) in the susceptibility to and severity of rheumatoid arthritis (RA) in Dutch patients. METHODS: A total of 319 consecutive RA patients, and a cohort of 90 female RA patients with detailed 12 yr follow-up were genotyped for the TNF-RI exon 1 (+36 A to G) and TNF-RII 3' UTR (+1690 T to C) polymorphisms. RESULTS: The frequencies of the TNF-RI and TNF-RII polymorphisms were determined in both patient groups and healthy controls, but no significant differences were found. To determine the relationship of these polymorphisms to disease severity, the extent of joint damage in the cohort of 90 female RA patients was analysed. No differences in severity were observed. CONCLUSION: These TNF-RI and TNF-RII polymorphisms were not found to be associated with susceptibility to or severity of RA in the Dutch population. PMID- 12730510 TI - Prevalence and clinical significance of anti-cyclic citrullinated peptide and antikeratin antibodies in palindromic rheumatism. An abortive form of rheumatoid arthritis? AB - OBJECTIVE: To analyse the prevalence and clinical significance of anti-cyclic citrullinated peptide antibodies (anti-CCP) and antikeratin antibodies (AKA) in patients with palindromic rheumatism (PR). METHOD: Sixty-three patients with PR were included: 33 were defined as pure or persistent PR at the time of serum test measurement, and 30 as associated PR, defined as patients with past history of PR who had developed persistent arthritis at the time of serum test: [21 with rheumatoid arthritis (RA)]. Sixty patients with early RA, and 80 with seronegative spondyloarthropathy were included as control groups. Anti-CCP were determined by a standardized ELISA test and AKA by indirect immunofluorescence in rat oesophagus. Clinical characteristics of these pure PR patients were compared according to the presence or absence of anti-CCP antibodies. A follow-up study was also performed. RESULTS: Anti-CCP were detected in 18 out of 32 (56.3%) patients with pure PR and 10 out of 30 (33.3%) with associated PR (38.1% in RA associated PR patients). AKA were detected in 12 patients out of 33, with pure PR (36.4%), and in 9 out of 30 with associated PR (30%) (33.3% in RA-associated PR patients). The prevalence of anti-CCP and AKA in the RA control group was 55% (not significantly different from the pure PR group) and 61.7% (with respect to pure PR patients, P=0.02), respectively. In the spondyloarthropathy group, the prevalence of anti-CCP and AKA was 2.5 and 3.8%, respectively (P<0.001 compared with pure PR patients). No significant clinical differences were observed between pure PR patients with and without CCP antibodies. CONCLUSIONS: Anti-CCP and, to a lesser extent, AKA, were found in a high proportion of patients with PR, suggesting that this syndrome is an abortive form of RA. The predictive value of these antibodies in PR, as markers of progression to an established RA, remains uncertain. PMID- 12730511 TI - Ultrasound guidance allows accurate needle placement and aspiration from small joints in patients with early inflammatory arthritis. AB - OBJECTIVES: To compare the accuracy of palpation-guided and high frequency ultrasound-guided needle placement in small joints and to develop a technique to obtain synovial fluid from these joints for diagnosis and research. METHODS: The accuracy of needle placement during palpation-guided proximal interphalangeal (PIP) or metacarpophalangeal (MCP) joint injection was assessed. This was compared with the accuracy of ultrasound-guided needle placement. A joint lavage technique was developed to obtain synovial fluid from these joints. RESULTS: Needle positioning was intra-articular in 59% of palpation-guided injections (6/12 PIP and 4/5 MCP joints). No fluid could be aspirated prior to injection. With ultrasound guidance, initial needle placement was intra-articular in 96% of cases (24/26 PIP and 27/27 MCP joints). Synovial fluid cells were lavaged from 63% of joints (19/25 PIP and 14/27 MCP joints). In only one case was a large effusion seen and this was aspirated directly. CONCLUSIONS: The use of high frequency ultrasound to guide needle placement within a small joint allows for significantly greater accuracy than a palpation-guided approach. When followed by lavage, synovial fluid cells and diluted synovial fluid can be obtained from the majority of small joints. This has important clinical and research implications. PMID- 12730512 TI - A novel polymorphism of the human APRIL gene is associated with systemic lupus erythematosus. AB - OBJECTIVE: We investigated the association of gene polymorphisms in APRIL, a new member of the TNF family, with systemic lupus erythematosus. METHODS: To detect polymorphisms of the human APRIL gene by exon-specific polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) analysis, we first determined the structure of the human APRIL gene. We designed exon-specific oligonucleotide primers according to the genomic DNA sequence of APRIL. All of the coding regions in exons of the APRIL gene were analysed by exon-specific PCR-SSCP in 148 SLE patients and 146 unaffected controls, then the nucleotide sequences of exons that displayed aberrant bands were determined. RESULTS: The human APRIL gene comprised at least six exons with five introns, spanning approximately 2.8 kilobases of the genomic DNA. By exon-specific PCR-SSCP, we identified two novel polymorphisms at codons 67 and 96. Both had amino acid substitutions: G67R and N96S respectively. Only the 67G allele was associated with SLE in 148 Japanese SLE patients, with allele frequency 0.662 compared with 0.575 for 146 unaffected controls (P=0.0302). The frequency of the individuals who possessed at least one 67G allele in SLE patients (91.9%) was significantly higher than that in the unaffected controls (80.1%) (P=0.0036). CONCLUSION: The 67G allele of APRIL may be a contributing factor in the pathogenesis of SLE. PMID- 12730513 TI - Glucose-6-phosphate isomerase is not a specific autoantigen in rheumatoid arthritis. AB - OBJECTIVE: To test the hypothesis that glucose-6-phosphate isomerase (GPI) is a novel autoantigen in RA. METHODS: Eighty-eight serum samples from 23 patients with rheumatoid arthritis (RA), 25 with Sjogren's syndrome, 20 with systemic lupus erythematosus and 20 healthy controls were tested by enzyme-linked immunosorbent assay (ELISA) using a commercially available, partially purified rabbit GPI as antigen. Beside each duplicate well containing antigen (10 micro g/ml), uncoated blocked duplicate wells (phosphate-buffered saline only) were included as controls for non-specific binding for every serum tested. We also examined antibodies binding to various polypeptides in the GPI preparation by immunoblotting in 73 of the sera. RESULTS: By ELISA, binding levels were low and there was no difference between serum from patients with RA, other rheumatic diseases and normal controls. By immunoblotting, antibodies binding to the GPI polypeptide were present in 70-80% of all groups tested. In addition, we showed that another polypeptide identified as phosphoglucomutase was also present in the preparation and reacted with human immunoglobulins. CONCLUSION: Our findings suggest that GPI is not a specific autoantigen in RA. PMID- 12730514 TI - Tissue factor in antiphospholipid syndrome: shifting the focus from coagulation to endothelium. PMID- 12730515 TI - Combined oral cyclosporin and methotrexate therapy in patients with rheumatoid arthritis elevates methotrexate levels and reduces 7-hydroxymethotrexate levels when compared with methotrexate alone. AB - OBJECTIVE: To study the pharmacokinetics of methotrexate (MTX) plus cyclosporin A (CSA) in patients with rheumatoid arthritis (RA). METHODS: On day 1 of the study, patients with RA receiving stable doses of MTX had blood and urine levels of MTX and its metabolite 7-hydroxymethotrexate (7-OH-MTX) measured post oral dosing of the drug. MTX was then discontinued and CSA therapy was started on day 8. On day 20, blood levels of CSA and CSA metabolites were measured post drug dosing. On day 23, MTX therapy was restarted and levels of MTX, CSA and their metabolites were again measured as described above. RESULTS: In the 30 patients, coadministration of CSA and MTX led to a 26% increase in mean peak plasma MTX concentration (P < 0.01), an 18% increase in the mean plasma MTX concentration area under the curve (AUC, P=0.01) and an 80% decrease in plasma 7-OH-MTX AUC (P < 0.01). In 13 patients receiving a 10 mg MTX dose, CSA reduced urinary 7-OH-MTX excretion by 87% (P < 0.01) without altering MTX excretion. MTX did not alter the pharmacokinetics of CSA or its metabolites. CONCLUSION: CSA may block oxidation of MTX to its relatively inactive metabolite, 7-OH-MTX, thereby potentiating MTX efficacy. PMID- 12730516 TI - Women with established rheumatoid arthritis perceive pain as the predominant impairment of health status. AB - OBJECTIVE: The aim of this study was to characterize perceptions of health and well-being in women with established rheumatoid arthritis (RA). METHODS: Women aged between 40 and 60 yr with RA for more than 3 yr, and who were receiving stable doses of anti-rheumatic drug therapy, were selected for study. The Arthritis Impact Measurement Scales 2 (AIMS2) was employed to quantify current health status impairments, the impairments that were attributed to RA, perceptions of current health status compared with other women of the same age, and perceptions of future health status. RESULTS: Fifty-eight patients were studied. The full range of 12 AIMS2 dimensions was used when quantifying impairments in health status. The dominant impairment was pain. Moreover, 88% of patients attributed current pain to RA. Fifty-two per cent perceived their health status to be fair, poor or very poor compared with other women of the same age. Twenty-five per cent expected poor future health status, compared with 10% who perceived poor current health status. Only one patient (2%) expected excellent future health status. CONCLUSION: In this study, a large majority of women perceived impairments of health status that they attributed to RA. Pain was perceived as the predominant impairment. Deterioration in health status was anticipated by many. The inclusion of patient perceptions of health status and disease impact as outcome measures in both cross-sectional and longitudinal clinical research protocols merits further study. PMID- 12730518 TI - Radiological progression in early rheumatoid arthritis after DMARDS: a one-year follow-up study in a clinical setting. AB - OBJECTIVE: To analyse the frequency and prognostic factors of radiographic progression in a series of Spanish patients with early rheumatoid arthritis (RA) after 1 yr of treatment with disease-modifying anti-rheumatic drugs (DMARDs). METHODS: Sixty patients (47 females, 13 males) with RA with a disease duration shorter than 2 yr [mean (s.d.) duration 9.5+/-6.6 months] were treated with the same therapeutic protocol using gold salts as the first DMARD and methotrexate as a second option, and were followed up for 1 yr. Radiographic progression in the hands and feet (total radiographic Larsen score and the erosion joint count) was used as the outcome variable. Clinical, laboratory, immunogenetic and radiographic data were obtained at study entry. Disease activity and response to therapy were measured at 6 and 12 months. RESULTS: Erosive disease was found in 21.7% of patients at baseline and in 38.3% after 1 yr. Although a substantial reduction in disease activity was observed during the 1 yr follow-up [disease activity score (DAS28) 5.8+/-0.8 at entry and 3.9+/-1.3 at 12 months, P < 0.001], the Larsen score rose from 1.9+/-3.3 to 5.6+/-9.8 after 1 yr. In 26.6% of patients, a raised erosion joint count was observed after 1 yr. Radiographic progression in the total joint radiographic damage (increase in Larsen score of >or=2) was observed in 36.6%. In the multivariate analysis, baseline pain [visual analogue scale (VAS)] and the presence of two copies of the shared epitope were associated with radiographic progression in the erosion joint count. Disease duration before study entry, VAS pain and Larsen score at baseline were significant predictors of radiographic progression in total damage (Larsen score). Baseline radiographic damage had the highest positive predictive value for progression. CONCLUSIONS: Radiographic progression was observed in up to 36.6% of patients with early RA after 1 yr of DMARD therapy in spite of a significant reduction in disease activity. Baseline factors, such as VAS pain, disease duration until DMARD therapy, damage score at baseline and the presence of two copies of the shared epitope, were associated with radiographic progression. PMID- 12730517 TI - Silicate antibodies in Danish women with silicone breast implants. AB - OBJECTIVES: To use a new immunological assay to evaluate silicate antibody levels in women with and without silicone breast implants (SBIs). METHODS: Women (n=186) were identified through Danish population-based registers and categorized into six groups defined by prior breast surgery (breast implantation/breast reduction/no breast surgery) and by the presence or absence of prior hospital diagnoses of soft-tissue rheumatism (muscular rheumatism, ICD-8 codes 717.90 and 717.99). The women underwent blood tests, a silicate antibody assay and a clinical examination. Severity of rheumatic symptoms/signs was scored from 1 (none) to 5 (severe). RESULTS: The level of silicate antibodies was not significantly different between the three groups with prior soft-tissue rheumatism (P > 0.5), with the lowest value among women with SBIs. Among women who had no prior diagnosis of soft-tissue rheumatism, silicate antibody levels were highest in women with SBIs (P < 0.01). No significant correlations were observed between silicate antibody levels and symptom severity scores. CONCLUSIONS: Silicate antibodies were not consistently associated with SBIs and were not correlated with rheumatic symptoms. The clinical relevance of these antibodies remains questionable. PMID- 12730519 TI - Fatigue in systemic lupus erythematosus: a randomized controlled trial of exercise. AB - OBJECTIVE: To test the efficacy of a graded aerobic exercise programme in treating fatigue in systemic lupus erythematosus. METHODS: Ninety-three patients with systemic lupus erythematosus without active disease in any major organ were randomized, using a minimization protocol, to 12 weeks of graded exercise therapy, relaxation therapy or no intervention. RESULTS: Analysis by intention to treat showed that 16 of the 33 (49%) patients in the exercise group rated themselves as 'much' or 'very much' better compared with eight out of 29 (28%) in the relaxation group and five out of 32 (16%) in the control group (chi2=8.3, df=2, P=0.02). Fatigue improved significantly on one out of three measures after exercise therapy and there was a trend for fatigue to improve on all measures after exercise. CONCLUSION: These findings support the use of appropriately prescribed graded aerobic exercise in the management of patients with fatigue and systemic lupus erythematosus. PMID- 12730520 TI - Homocysteine levels in polymyalgia rheumatica and giant cell arteritis: influence of corticosteroid therapy. AB - OBJECTIVES: It has been suggested that patients with giant cell arteritis (GCA) may share a common pathway with atherosclerosis. Furthermore, patients with GCA and polymyalgia rheumatica (PMR), in addition to advanced age, are treated for prolonged periods of time with corticosteroids, a factor that can also accelerate atherosclerosis. Hyperhomocysteinaemia is considered an independent risk factor for atherosclerosis, and might play a role in ischaemic manifestations that occur with a variable frequency during the course of GCA. The purposes of the present study were: (i). to analyse the plasma levels of homocysteine in patients with GCA and PMR, (ii). to determine the influence of corticosteroid therapy on the homocysteine levels and (iii). to analyse if the levels of homocysteine may predict the development of ischaemic complications in patients with GCA. METHODS: Plasma homocysteine concentration was measured in 56 patients with active PMR/GCA (17 GCA and 39 isolated PMR) before steroid treatment and 23 healthy age-matched volunteers were used as controls. The total plasma homocysteine level was quantified using a fluorescent polarization immunoassay. RESULTS: Homocysteine concentrations were higher in PMR and GCA patients than age-matched controls (P < 0.05). Patients with GCA had slightly higher levels of plasma homocysteine than those with isolated PMR (13.6+/-4.3 vs 12.7+/-3.1 micromol/l, P=0.6). In 30 of these patients (12 GCA and 18 PMR) a second measurement of homocysteine concentration was done when they were in clinical remission with steroid treatment. The post-treatment levels of homocysteine were significantly increased in GCA rather than in PMR patients. In 13 patients with homocysteine levels above the normal upper limit of our laboratory, therapy with folic acid and/or vitamin B12 was started. After 3 months of vitamin supplements, the homocysteine concentration significantly decreased from 19.2+/-3.1 to 13.6+/-3.2 micromol/l (P=0.001). Such decrease was less marked in the PMR than in GCA patients. Ten out of the 17 patients with GCA had ischaemic manifestations of the disease. The levels of homocysteine were slightly higher in GCA patients with ischaemia than in those without ischaemic manifestations, although the difference did not reach statistical significance (15+/-4.9 vs 11.6+/-1.9 micromol/l, P=0.46). CONCLUSIONS: Patients with active PMR and GCA had elevated plasma concentrations of homocysteine. Corticosteroid therapy significantly increased such levels, especially in GCA patients. Treatment with supplements of folic acid and/or vitamin B12 reduced the homocysteine concentrations. These data support the hypothesis that patients with GCA (and to a lesser extend PMR patients) may share a common pathway with atherosclerosis and suggest a new atherogenic mechanism of corticosteroids. PMID- 12730521 TI - Concomitant septic and gouty arthritis--an analysis of 30 cases. AB - OBJECTIVES: To analyse the clinical features and outcomes of gouty patients with concomitant septic arthritis in a medical centre. METHODS: From the hospital database, we collected 30 hospitalized cases with concomitant septic arthritis and gouty arthritis from 1987 to 2001. All patients had positive bacterial culture and monosodium urate crystals in the affected joints. Medical records of the patients were analysed in detail. RESULTS: The mean age of patients was 52.8+/-12.5 yr. One-third of patients were afebrile at presentation, 30% had a normal blood leucocyte count and 10% had a synovial fluid leucocyte count less than 6000/mm3. The knee joint was the most common site of involvement, followed by the ankle, shoulder and wrist joints. Most patients had long-standing disease and subcutaneous tophi. Subcutaneous tophi rupture with secondary wound infection is the most common route of infection. Causative micro-organisms were Staphylococcus aureus (16 cases, 7 of whom were oxacillin-resistant), Streptococcus sp. (5 cases), Pediococcus sp. (1 case), and Gram-negative bacilli (9 cases). Fourteen patients received surgical debridement, among them two patients had an arthrodesis owing to severe joint destruction and one received above-knee amputation. Two patients died. One died of septic complications and the other died of acute myocardial infarction. CONCLUSIONS: Septic arthritis coexistent with gout presented a diagnostic difficulty. An early diagnosis requires a high level of suspicion. Prompt aspiration and analysis of the synovial fluid is imperative, regardless of the absence of fever or leucocytosis. Culture of the aspirated synovial fluid is warranted in gouty attack, even when it has a low white cell count or the Gram stain reveals no organisms. PMID- 12730522 TI - Referred sensations in patients with complex regional pain syndrome type 1. AB - OBJECTIVES: This study sought to explore and characterize referred sensations (RS) in patients with complex regional pain syndrome (CRPS) type 1 and test the hypothesis that pain in CRPS is associated with central sensory changes. METHODS: Subjects underwent standardized neurological examination involving light touch, pinprick and vibration sense with eyes closed and then with eyes open. The subjects described the location and sensation emanating from the stimulated site and whether they experienced any sensations (similar or different) elsewhere. RESULTS: Five of 16 subjects recruited demonstrated RS. These were experienced in real time, were modality specific (touch and pinprick) and were located on the body part immediately adjacent, on Penfield's cortical homunculus, to the stimulated site. The RS were diminished or absent when the subject visualized the stimulated area. They disappeared when stimulation ceased and on clinical improvement. CONCLUSIONS: This is the first report of RS in CRPS and provides further evidence of central reorganization in what was previously thought to be a peripheral disorder. PMID- 12730523 TI - Therapeutic management of extrahepatic manifestations in patients with chronic hepatitis C virus infection. PMID- 12730524 TI - Surface microscopy for discriminating between common urticaria and urticarial vasculitis. AB - BACKGROUND: Urticarial vasculitis is a clinicopathological entity that overlaps with common urticaria, and biopsy is required for differentiation between them. OBJECTIVE: To determine, for the first time, if skin surface microscopy can aid in the clinical differentiation between common urticaria and urticarial vasculitis in daily practice. PATIENTS AND METHODS: Lesions in 20 consecutive patients with a clinical diagnosis of urticaria were studied by biopsy and skin surface microscopy (10x dermoscope) after covering the lesions with olive oil. Lesions were photographed with Dermaphot equipment. A biopsy was taken from all patients. Statistical analysis included Fisher's exact test and Cohen kappa statistics (intra-observer reproducibility). RESULTS: Two dermoscopic patterns were observed: (i). a red-lined vascular pattern (17/20 patients); and (ii). a purpuric globular pattern (3/20 patients). Leucocytoclastic vasculitis was demonstrated histologically in all lesions presenting purpuric globules (3/3) but in none of the lesions presenting a dermoscopic red-lined pattern (P < 0.0008). The intra-observer reproducibility for scoring the red lines and purpuric globules was excellent (kappa=0.8). CONCLUSION: The results of this pilot study suggest that skin surface microscopy, using a 10x dermoscope, detects purpuric globules in urticarial lesions, and that purpuric globules indicate underlying leucocytoclastic vasculitis. PMID- 12730525 TI - Health problems following Campylobacter jejuni enteritis in a Lancashire population. AB - OBJECTIVES: Campylobacter jejuni enteritis can lead to musculoskeletal, neuropathic or other health sequelae. We investigated the coexistence, seasonal occurrence, strain-type associations and impact on work capacity of different health problems following C. jejuni enteritis in a Lancashire population during 1999 and 2001. METHODS: A semistructured questionnaire was used to characterize health problems that occurred in the community after laboratory-confirmed episodes of C. jejuni enteritis. The questionnaire was posted to all adults in the Preston and Chorley area who developed C. jejuni enteritis in 1999 or 2001. All Campylobacter isolates from this population were serotyped. RESULTS: Several types of sequelae occurred consistently in both years, including the coexistence of musculoskeletal and neuropathic problems. There was no evidence of C. jejuni strain-type associations or seasonal preponderance for any type of sequela. The overall health impact of C. jejuni enteritis, as measured by workdays lost, was high in this population. CONCLUSIONS: A variety of health problems occur consistently following C. jejuni enteritis and substantially increase morbidity due to campylobacteriosis in the community. PMID- 12730526 TI - Psoriasis vulgaris and psoriatic arthritis share a 100 kb susceptibility region telomeric to HLA-C. AB - OBJECTIVE: To map the locus for susceptibility to psoriasis in patients with psoriatic arthritis. METHODS: Seventy-four patients with psoriatic arthritis and 95 patients with psoriasis vulgaris were included in this study. Polymorphic genes and microsatellite markers centromeric (C1_2_5) and telomeric (C1_4_4, OTF3, HCR and the corneodesmosin gene) to HLA-C were studied in an association analysis. Typing was also performed on a control population of 104 matched donors. RESULTS: The allele Cw*0602 was associated both with psoriasis [49 vs 21%, Pc<0.0003; odds ratio (OR)=3.6, aetiological factor (AF)=0.72] and with psoriatic arthritis (62 vs 21%, Pc<0.000001; OR=6.1, AF=0.83). In psoriatic patients a susceptibility region telomeric to HLA-C that includes C1_4_4 (56 vs 22%, Pc<0.0002; OR=4.39, AF=0.77), OTF3 (85 vs 60%, Pc<0.0002; OR=3.7, AF=0.73) and HCR (63 vs 26%, Pc<0.00001; OR=3.8, AF=0.74) was observed. In psoriatic arthritis patients the susceptibility region was delimited by HLA-C and C1_4_4 (384 allele, 56 vs 22%, Pc<0.0002; OR=4.37, AF=0.77). CONCLUSIONS: Comparing the susceptibility regions associated with the two diseases, an overlapping interval of 100 kb between HLA-C and OTF3, which might contain the psoriasis gene, can be defined. PMID- 12730528 TI - A cross-sectional comparison of three self-reported functional indices in scleroderma. AB - OBJECTIVES: In scleroderma, outcome measures such as skin score provide only limited information about the functional impact of the disease. The requirement for validated and convenient instruments that reliably reflect disease morbidity is now recognized. This study compares the Disability Index of the Health Assessment Questionnaire (HAQ-DI) with two more recently developed scleroderma specific tools: scleroderma-visual analogue scales (scleroderma-VAS) and the UK scleroderma Functional Score (UKFS). In addition, the use of clinical and laboratory measures as predictors of disease severity have been examined. METHODS: One hundred and fifteen consecutive patients were studied. Subjects completed the 20-item HAQ-DI, the scleroderma-VAS and a questionnaire related to hand and muscle function (UKFS). Clinical details, measurement of maximal hand spread, fist-closure and investigations for internal organ involvement were recorded. RESULTS: Over 68% of patients with diffuse disease had moderate to severe disease on the UKFS, compared with 44% with limited disease. The mean UKFS in diffuse disease was 14.7 (s.d. 9.1) and 10.6 (s.d. 8.5) in the limited subset (P=0.02). The mean HAQ-DI in diffuse disease was 1.23 (s.d. 0.77) and 0.79 (s.d. 0.75) in the limited subset (P=0.005). The HAQ-DI showed significant correlation with UKFS (r=0.9; P < 0.001). Several clinical and laboratory measures were associated with higher HAQ-DI and UKFS. CONCLUSIONS: This is the first comparative study of the UKFS and the HAQ-DI. These data show a strong correlation between assessment methods. Higher scores correlated with clinical and laboratory indicators of severe disease. Used together, these inexpensive tools assess general and organ-specific symptoms, as well as functional limitation. PMID- 12730527 TI - MRL/lpr lupus-prone mice show exaggerated ICAM-1-dependent leucocyte adhesion and transendothelial migration in response to TNF-alpha. AB - OBJECTIVE: Endothelial activation and dysfunctional leucocyte-endothelial interactions are thought to play key roles in the pathogenesis of systemic lupus erythematosus (SLE). The object of this study was to investigate directly the effect of increased endothelial adhesion molecule expression on leucocyte endothelial cell interactions, using the MRL/lpr mouse model. METHODS: Leucocyte rolling, arrest and transendothelial migration were quantified in the cremaster muscle microcirculation of 20-week-old MRL/lpr mice, using intravital microscopy. Endothelial adhesion molecule expression was quantified using intravenously injected radiolabelled monoclonal antibodies. RESULTS: Basal expression of intercellular adhesion molecule 1 (ICAM-1) by cremaster endothelium was 2-fold greater in MRL/lpr than in MRL/++ mice (P<0.05). There was a 1.6-fold increase in expression of vascular adhesion molecule 1 (VCAM-1), but no increase in E selectin or P-selectin expression. Following intrascrotal injection of saline, no difference was detected in leucocyte-endothelial interactions between MRL/lpr and control MRL/++ mice. In contrast, intrascrotal injection of tumour necrosis factor alpha (TNF-alpha) (2 h test period) led to significantly increased numbers of adherent and extravasated leucocytes in MRL/lpr (5.98+/-0.71 and 5.45+/-0.34 leucocytes per 100 micro m vessel segment respectively) compared with MRL/++ mice (3.63+/-0.26 and 2.97+/-0.24 respectively, each P<0.05). Treatment of TNF-alpha stimulated mice with anti-ICAM-1 F(ab')2 (YN1) abolished the difference between MRL/lpr and MRL/++ mice, whereas a negative control anti-DNP F(ab')2 had no effect. CONCLUSIONS: MRL/lpr lupus-prone mice show exaggerated ICAM-1-dependent leucocyte-endothelial interactions in response to TNF-alpha. Increased leucocyte endothelial interactions due to endothelial priming could contribute to the clinical link between infection and flares of lupus disease activity. PMID- 12730529 TI - Genetic associations between frozen shoulder and tennis elbow: a female twin study. AB - OBJECTIVES: To estimate the heritability of frozen shoulder (FS) and tennis elbow (TE) and to examine the two disorders for possible genetic or environmental associations. METHODS: Self-reporting questionnaire data on risk factors for, and a physician diagnosis of, FS or TE were obtained from 865 monozygotic (MZ) and 963 dizygotic (DZ) unselected female twin pairs aged between 20 and 76 yr registered with the St Thomas' UK Adult Twin Registry. The heritability of each disorder was estimated in a classic twin study. The association between FS and TE was then explored by log-linear modelling comparing MZ with DZ individuals and twin pairs for the presence of both disorders. RESULTS: The prevalence of FS and TE were 11.6 and 16.7%, respectively. A heritability of 42% was estimated for FS and 40% for TE after adjusting for age. There was no confounding by environmental risk factors. Log-linear modelling demonstrated FS and TE, independently, to be associated within members of a twin pair and confirmed a stronger association in MZ than DZ pairs. In addition the two disorders occurred together 2-3 times more frequently in individuals than would be expected by chance. However, there was no association between FS and TE across members of a twin pair, implying no evidence for a shared genetic component to the two disorders. CONCLUSIONS: Genetic factors are implicated in the aetiology of both frozen shoulder and tennis elbow but are independent of each other. The two disorders occur together 2-3 times more frequently than by chance in individuals. However, the association is most likely mediated by individual-specific environmental factors common to the two conditions and not by a common genetic susceptibility. PMID- 12730530 TI - The blossoming of evidence-based clinical rheumatology: The Arthritis Research Campaign's Clinical Trials Collaboration in association with the MRC Clinical Trials Unit, BSR and BOA. PMID- 12730531 TI - Elements of fibromyalgia in an open population. AB - OBJECTIVE: To examine the nosological concept of fibromyalgia in the general population. METHODS: A postal survey of rheumatic pain and non-specific bodily complaints was sent to all 3174 German female residents of Bad Sackingen, Germany, aged 35 to 74 yr. A stratified random sample of 653 subjects was further examined in a clinical survey. RESULTS: On the population level the point prevalence of chronic widespread pain was 13.5%. In the clinical survey, tender point count was associated not only with the extent of rheumatic pain, but also independently with the extent of bodily complaints. Subjects with no history of rheumatic pain but with non-specific bodily complaints had as many positive tender points as subjects without bodily complaints but with a history of rheumatic pain. Subjects could be identified who met the tender point criterion of the ACR without a history of widespread pain. Multivariate analyses demonstrated that some symptoms carry a risk for positive tender points (low physical mobility, pain, bodily complaints) and some for chronic widespread pain (poor health status, catastrophizing, emotional reactions, low energy level, sleep disturbances) that are independent of each other and of age. CONCLUSIONS: The results do not only question the relevance and specificity of a history of widespread pain in diagnosing fibromyalgia, but also the concept of fibromyalgia as a distinct rheumatological disorder. The results support the concept of fibromyalgia as part of a wider spectrum of dysfunctional syndromes. PMID- 12730532 TI - Three-monthly ibandronate bolus injection offers favourable tolerability and sustained efficacy advantage over two years in established corticosteroid-induced osteoporosis. AB - OBJECTIVE: Corticosteroids are widely prescribed, although treatment-related side effects are common. Of these adverse events (AEs), osteoporosis is considered the most serious. Currently, oral bisphosphonates are the standard treatment for corticosteroid-induced osteoporosis (CIO). However, intermittent intravenous (i.v.) therapy may have advantages, including lack of gastrointestinal AEs, improved bioavailability and increased compliance. This study investigated the efficacy and safety of 3-monthly i.v. ibandronate bolus injections in patients with established CIO. The results from a planned 2-yr interim analysis are reported. METHOD: In this controlled, prospective, open-label, parallel-group study, 104 patients (49 men and 55 women) with established CIO (mean T-score < 2.5 s.d. at the lumbar spine (L2-L4) received daily calcium (500 mg) plus either 3-monthly i.v. ibandronate (2 mg) bolus injections or oral daily alfacalcidol (1 micro g). The primary end-point was bone mineral density (BMD) change at the lumbar spine, femoral neck and calcaneus after 24 months. RESULTS: Compared with oral daily alfacalcidol, i.v. ibandronate produced significantly superior gains in mean (+/-s.d.) BMD at the lumbar spine (2.2+/-3.1 vs 11.9+/-7.4%; P<0.001), femoral neck (1.3+/-1.8 vs 4.7+/-4.0%; P<0.001) and calcaneus (7.6+/-3.8 vs 15.5+/-10.7%; P<0.0001) after 2 yr. Consistent with these BMD gains and, although the study was not powered for fractures, a trend towards a reduction in vertebral fractures and greater back pain relief was seen in the ibandronate group. The overall incidence of AEs was similar in the two treatment arms. CONCLUSIONS: Three-monthly i.v. ibandronate bolus injections are significantly superior to alfacalcidol in the treatment of CIO. These data confirm the potential of ibandronate for the treatment of osteoporosis associated with corticosteroid use. The ease of administration, lack of AEs and good compliance associated with intermittent i.v. ibandronate make it a potentially valuable alternative to oral bisphosphonate therapy for the treatment of CIO. PMID- 12730534 TI - Quantifying the burden of emotional ill-health amongst patients referred to a specialist rheumatology service. AB - OBJECTIVES: (1) To determine the prevalence of emotional disorders (DSM IV depression, anxiety and panic disorders) amongst patients referred to a rheumatology out-patient service and the proportion of these detected by the rheumatologist. (2) To test the hypotheses that emotional disorders are associated with (i) broad categories of rheumatological diagnosis (systemic, inflammatory vs non-systemic, non-inflammatory), (ii) female gender, (iii) greater symptom burden and disability and (iv) markers of socio-economic deprivation. METHODS: A cross-sectional study was made of consecutive newly referred attenders at a hospital-based, regional rheumatology service. Emotional disorders, pain, health status and socio-economic factors were assessed by questionnaire. The letter to the referrer was scrutinized for the rheumatological diagnosis and mention of emotional disorder. RESULTS: A total of 256 patients were eligible and 203 (79%) participated. The sample was 69% female, had a mean age of 50 yr and 68 patients (33.5%) had one or more emotional disorders. Only a minority were detected. There was no association with type of rheumatological diagnosis. Patients with an emotional disorder were more likely to be female (81 vs 62%; P<0.007), to report more pain (mean Visual Analogue Score 70 vs 50 mm, P<0.001), a greater number of somatic symptoms (median 3 vs 1, P<0.001) and greater disability (median Health Assessment Questionnaire 1.1 vs 0.5, P<0.001). Emotional disorders were also associated with some, but not all, measures of lower social and economic status and life dissatisfaction. CONCLUSIONS: Emotional disorder is present in one-third of new rheumatology referrals. The course, causation and management of this important component of rheumatological illness merit further attention. PMID- 12730535 TI - Cardiovascular risk in systemic lupus erythematosus--evidence of increased oxidative stress and dyslipidaemia. AB - OBJECTIVES: Systemic lupus erythematosus (SLE) is associated with severe and premature cardiovascular disease, which is not explained by traditional risk factors alone. This study aimed to investigate markers of oxidative stress, lipid metabolism and inflammation as potential cardiovascular risk factors in women with SLE. METHODS: Venous blood samples were taken from 53 female Caucasian patients with SLE and from healthy age- and sex-matched controls. Samples were analysed for markers of oxidative stress, lipid metabolism [including low-density lipoprotein (LDL) subfraction profile] and C-reactive protein (CRP). RESULTS: Female SLE patients had an atherogenic lipid profile characterized by raised total cholesterol and triglycerides, and the presence of small, dense LDL subfractions compared with healthy controls. These changes were associated with increased oxidative damage and a moderately raised CRP. CONCLUSIONS: The results provide evidence for free radical and inflammatory activity in SLE and suggest potential targets to reduce the risk of cardiovascular disease in these patients. PMID- 12730536 TI - Can diagnostic triage by general practitioners or rheumatology nurses improve the positive predictive value of referrals to early arthritis clinics? AB - OBJECTIVES: To determine whether diagnostic triage by general practitioners (GPs) or rheumatology nurses (RNs) can improve the positive predictive value of referrals to early arthritis clinics (EACs). METHODS: Four GPs and two RNs were trained in the assessment of early inflammatory arthritis (IA) by four visits to an EAC supervised by hospital rheumatologists. Patients referred to one of three EACs were recruited for study and assessed independently by a GP, an RN and one of six rheumatologists. Each assessor was asked to record their clinical findings and whether they considered the patient to have IA. Each was then asked to judge the appropriateness of the referral according to predetermined guidelines. The rheumatologists had been shown previously to have a satisfactory level of agreement in the assessment of IA. RESULTS: Ninety-six patients were approached and all consented to take part in the study. In 49 cases (51%), the rheumatologist judged that the patient had IA and that the referral was appropriate. The assessments of GPs and RNs were compared with those of the rheumatologists. Levels of agreement were measured using the kappa value, where 1.0 represents total unanimity. The kappa value was 0.77 for the GPs when compared with the rheumatologists and 0.79 for the RNs. Significant stiffness in the morning or after rest and objective joint swelling were the most important clinical features enabling the GPs and RNs to discriminate between IA and non-IA conditions. CONCLUSION: Diagnostic triage by GPs or RNs improved the positive predictive value of referrals to an EAC with a degree of accuracy approaching that of a group of experienced rheumatologists. PMID- 12730537 TI - Acceptability and compliance with hip protectors in community-dwelling women at high risk of hip fracture. AB - OBJECTIVES: To determine acceptability and compliance with hip protectors in women at high risk of hip fracture who are living independently in the community. METHODS: Women aged 65 yr and over referred for open access bone densitometry who had femoral neck osteoporosis and a high risk of falling were asked to wear hip protectors. RESULTS: Eighty five women fulfilled the inclusion criteria of whom 32 (38%) found the hip protectors acceptable and agreed to participate. Reasons given by the remaining 53 (62%) for not finding the hip protectors acceptable included discomfort on wearing, dislike of their personal appearance with the hip protectors on, and disagreement about their fracture risk. Participants were more likely to have a family history of osteoporosis (47 vs 26%, respectively) and hip fracture (16 vs 8%) compared with non-participants. At 12 months only about half of the subjects were wearing hip protectors daily. CONCLUSIONS: Our findings suggest that only a minority of community-dwelling women at high risk of hip fracture will wear hip protectors to reduce fracture risk. Their use should be restricted to highly motivated women who should be carefully identified. PMID- 12730539 TI - Progression of peripheral joint disease in psoriatic arthritis: a 5-yr prospective study. AB - OBJECTIVE: To assess the evolution of disease subgroups and the frequency of progression of peripheral joint disease in a prospectively studied cohort of patients with psoriatic arthritis (PsA). METHODS: The cohort was identified as the first consecutive 100 patients attending a psoriatic arthritis clinic and who had been the subject of a previously published cross-sectional retrospective study. Nine of the 100 patients had died, three declined follow-up and one could not be traced. The remaining 87 patients (49 females, 38 males) completed the study proforma at a median follow-up interval of 65 months (range 39-90). An analysis of initial plasma viscosity compared with rates of progression of joint score was performed. RESULTS: Eighteen patients changed subgroup; 11 had an increase in the number of joints involved, six a decrease, and one changed from an oligoarticular pattern to predominant spondylitis. Within the polyarticular group 37/51 patients had an increase in the number of joints involved. For the whole population, there were significant increases in the number of joints involved (median 6 vs 11, P < 0.001 Wilcoxon signed rank) and Health Assessment Questionnaire scores (median 0.375 vs 0.5, P < 0.001). The median rate of joint progression was 0.42 peripheral joints per year (range 0-7.2). However, the rate of peripheral joint involvement was highest in the first year of arthritis (median 4.0 joints/yr) as measured in 13 patients who had onset within 12 months of baseline assessment. There were no significant differences in skin and nail scores although nine more patients had developed nail disease. There was a significant correlation between the initial viscosity and rate of progression of joint damage (Spearman correlation, P < 0.011). CONCLUSIONS: Peripheral joint disease is progressive in the majority of patients with PsA and reinforces the need for effective monitoring and treatment. PMID- 12730538 TI - Cerebral imaging by magnetic resonance imaging and single photon emission computed tomography in systemic lupus erythematosus with central nervous system involvement. AB - OBJECTIVE: To assess the significance of magnetic resonance imaging (MRI) and single photon emission computed tomography (SPECT) abnormalities in patients with systemic lupus erythematosus (SLE). METHODS: Forty-four patients with SLE were retrospectively analysed. Patients were classified into three groups [1 and 2: patients with central nervous system (CNS) manifestations before and after starting high-dose steroid therapy, respectively; 3: patients without CNS manifestations. MRI was performed in all 44 patients and SPECT in 31. RESULTS: Abnormal findings in MRI were found in 19 patients. MRI abnormalities were significantly more frequent in patients with CNS manifestations than in those without [71 vs 17%, odds ratio (OR) 11.9, confidence interval (CI) 2.8-49.9, P=0.0003]. After the initiation of steroid therapy, patients with CNS manifestations also had an increased frequency of abnormal MRI. No correlation was found between SPECT findings and CNS manifestations. However, patients with CNS manifestations after starting steroids showed a markedly increased frequency of abnormal MRI and SPECT compared with those without CNS manifestations (80 vs 7%; OR 56, CI 4.4-719, P=0.0003). The positive predictive value of abnormality in both techniques in developing CNS manifestations after starting steroids was 89%. CONCLUSION: MRI findings correlated with CNS manifestations in SLE. Where there is a high suspicion of CNS involvement, the combination of MRI and SPECT may be useful in predicting CNS manifestations after starting steroid therapy. PMID- 12730540 TI - Haemoglobin-derived iron-dependent hydroxyl radical formation in blood-induced joint damage: an in vitro study. AB - OBJECTIVE: It has been reported that joint bleeds cause cartilage damage and that the combination of red blood cells (RBC) plus mononuclear cells (MNC) causes the adverse effects. The present study is to elucidate the mechanism by which blood, as present in whole blood, may cause this cartilage damage. METHODS: Human cartilage samples were cultured for 4 days in the presence of 50% whole blood, isolated MNC plus RBC, CD14+ cells (monocytes/macrophages) plus RBC, or lysed RBC with interleukin 1beta (IL-1beta; a major catabolic product of activated monocytes/macrophages). Antioxidants were used to investigate the involvement of oxidative stress. A subsequent 12-day culture period in the absence of additions is referred to as the recovery period. Changes in cartilage proteoglycan synthesis were determined at days 4 and 16. RESULTS: Cartilage cultured in the presence of whole blood, MNC plus RBC, or monocytes/macrophages plus RBC resulted in a prolonged inhibition of proteoglycan synthesis (>90% inhibition at day 16; all three P<0.05). Lysed RBC together with IL-1beta also induced prolonged inhibition of proteoglycan synthesis (>56% of controls, P<0.05). Dimethylsulphoxide (DMSO), scavenging hydroxyl radicals, could reverse the inhibition of cartilage proteoglycan synthesis. CONCLUSIONS: Based on these results we hypothesize that IL-1beta produced by activated monocytes/macrophages increases the production of hydrogen peroxide by chondrocytes. This in combination with haemoglobin-derived iron from the RBC will result in the formation of hydroxyl radicals in the vicinity of chondrocytes. This mechanism may result in chondrocyte damage and as such be involved in blood-induced cartilage damage. PMID- 12730541 TI - Association of birth weight with osteoporosis and osteoarthritis in adult twins. AB - OBJECTIVES: Twin studies present a unique opportunity to examine the association of birth weight with adult life phenotypes in a design that naturally accounts for maternal factors and a range of early environmental factors, which might potentially bias the association. In this study, we explored the association of birth weight with osteoporosis (OP) and osteoarthritis (OA), in a large national cohort of female twins. METHODS: Intra-pair differences between the reported birth weight of the twins (n=4008) were examined for an association with: (i) intra-pair differences in bone mineral density (BMD) and bone mineral content (BMC) at the lumbar spine, hip and forearm using linear regression; and (ii) osteoarthritis status in pairs discordant for radiographic disease at the hand, hip and knee using matched logistic regression. The confounding influences of height and weight were taken into account. RESULTS: The mean age of the twins was 47.5+/-12.3 yr. Intra-pair differences in birth weight were significantly associated with BMD at the spine (P=0.047), total hip (P=0.016) and femoral neck (P<0.001), but not at the forearm (P=0.245). These were entirely explained by the birth weight association with height and weight. The associations of intra-pair differences in birth weight and BMC were highly significant (P<0.001) at all sites, but were partly explained by adjustment for adult height and weight. We found no clear association between intra-pair birth weight differences and OA in twins discordant for any of the radiographic OA phenotypes at any site. CONCLUSIONS: Bone mass and especially BMC are highly associated with birth weight. These associations are accounted for mainly by environmental factors that are independent of maternal factors such as gestational age, maternal smoking and nutrition, and are largely mediated by skeletal size and particularly adult height. Birth weight does not appear to be a major influence on the later development of radiographic OA in women. PMID- 12730543 TI - Predominant cellular immune response to the cartilage autoantigenic G1 aggrecan in ankylosing spondylitis and rheumatoid arthritis. AB - OBJECTIVES: Based on their HLA association, both ankylosing spondylitis (AS) and rheumatoid arthritis (RA) seem to be T-cell-driven diseases in which the autoantigens remain to be defined. One possible autoantigen is the G1 domain of aggrecan, the major cartilage proteoglycan. In BALB/c mice immunized with this protein, spondylitis and erosive polyarthritis have been reported. Immune reactivity to the G1 has been described in patients with RA and AS in an earlier study. Using novel and more sensitive techniques and relevant controls we sought to define the role of G1 as an autoantigen more precisely and to extend the specific analyses to the peptide level. METHODS: Peripheral blood (PB) mononuclear cells (MNC) from 47 AS patients, 22 RA patients and 20 healthy normal controls were exposed in vitro for 6 h to the cartilage-derived autoantigens G1, human cartilage (HC) gp-39 and collagen II. Synovial fluid (SF) MNC from seven AS and four RA patients were similarly analysed. Furthermore, PB MNC of 15 AS and 10 RA patients were examined with overlapping 18-mer peptides covering the whole G1 protein to identify the immunodominant epitopes. T cells were stained by monoclonal antibodies directed against the surface markers CD4, CD69 and against the intracellular cytokines interferon-gamma (IFN gamma), tumour necrosis factor alpha (TNF alpha), interleukin 4 (IL-4) and IL-10. The percentage of reactive T cells was quantified by flow cytometry. RESULTS: After antigen-specific stimulation with the G1 protein, the CD4+ T cells of 30 AS patients (61.7%) and of 12 RA patients (54.5%) secreted significant amounts of IFN gamma and TNF alpha, while, in contrast, only 10% of the normal controls showed a response (P < 0.05). The synovial CD4+ T cells of five AS (71.5%) and of all four RA patients showed antigen-specific responses to the G1. In contrast, stimulation with HC gp 39 and collagen II showed no significant IFN gamma and TNF alpha secretion of MNC in all groups. Several G1-derived T-cell epitopes were identified as immunodominant in PB MNC of AS and RA patients and were partly overlapping. CONCLUSIONS: These data show that a cellular immune response to G1 is present in most AS and RA patients. G1-immunodominant epitopes were identified. The relevance of this finding for the pathogenesis of AS and RA remains to be established. PMID- 12730544 TI - The prevalence of patients with rheumatoid arthritis in the West Midlands fulfilling the BSR criteria for anti-tumour necrosis factor therapy: an out patient study. AB - OBJECTIVES: There are currently two anti-tumour necrosis factor (anti-TNF) therapies licensed for treatment of rheumatoid arthritis (RA). A British Society for Rheumatology (BSR) working party defined criteria for patients that would be suitable for such treatment. The aim of this study was to determine the prevalence of these patients attending rheumatology out-patient departments across the West Midlands. METHODS: Data were collected over a 2-week period in adult out-patient departments of 12 centres. A questionnaire was completed at each patient review. Disease activity scores (DAS-28) were recorded for those who had failed methotrexate treatment and at least one other disease-modifying anti rheumatic drug (DMARD) in the absence of contraindications to anti-TNF therapy. Information was also collected on the number of DMARDs failed and the use of steroid therapy. RESULTS: A total of 1441 patients with RA were assessed; 177 (12.3%) patients had failed methotrexate and at least one other DMARD. Of these, 19 had contraindications to the use of anti-TNF therapy. In the remaining 158 patients (11%), 80 (5.6%) had a DAS-28 score of >5.1, thus fulfilling BSR criteria for use of anti-TNF therapy. Those with a DAS-28 score of < or = 5.1 were significantly more likely to have been taking steroids compared with those with a DAS-28 score >5.1 (68.2 and 49.3%, respectively, P=0.024). CONCLUSIONS: Of patients with RA attending adult rheumatology out-patient clinics in the West Midlands, 5.6% would meet BSR criteria for use of anti-TNF therapy. Eligibility may be affected by steroid use. PMID- 12730545 TI - Association of specific interleukin 1 gene cluster polymorphisms with increased susceptibility for Behcet's disease. AB - OBJECTIVE: The aim of this study was to investigate if the inheritance of specific polymorphisms of interleukin 1 (IL-1) A, IL-1B and IL-1 receptor antagonist (IL-1RN) genes could affect the susceptibility to Behcet's disease (BD). METHODS: A total of 132 BD patients and 105 healthy controls were genotyped for IL-1A -889, IL-1B -511, -35, +5810, +5887, and IL-1RN +8006, +8061, +9589, +11,100 single nucleotide polymorphisms, and IL-1RN 86-bp variable number of tandem repeat polymorphism. chi 2-analysis was used to compare the allele and genotype frequencies of the cases and controls. IL-1A and IL-1B haplotypes were reconstructed using the Phase program. RESULTS: Inheritance of the C allele of the IL-1A -889 polymorphism was associated with BD (OR=2.0, P=0.01) and inheritance of the IL-1A -889C/IL-1B +5887T haplotype was identified as an increased risk for BD. The IL-1A -889 and IL-1B +5887 CC/TT combined genotype was significantly more observed in BD cases than in controls (57.5 vs 38.1%, OR=2.2, P=0.003). No association with BD was found for other investigated polymorphisms in the IL-1B and IL-1RN genes. CONCLUSION: Susceptibility to BD is increased in individuals carrying both the IL-1A -889C and IL-1B +5887T haplotype. Individuals who are both homozygous CC at IL-1A -889 and TT at IL-1B +5887 appear to have twice the risk of developing BD as individuals having other IL-1A -889/IL-1B +5887 genotypes. PMID- 12730546 TI - Immediate effects of adhesive tape on pain and disability in individuals with knee osteoarthritis. AB - OBJECTIVES: To evaluate the effects of two knee taping techniques, therapeutic tape and neutral tape, on pain and observed disability in symptomatic generalized knee osteoarthritis (OA). METHODS: Using a within-subjects study design, 18 participants were tested under three conditions in random order: untaped, wearing therapeutic knee tape and wearing neutral knee tape. Outcome measures included assessment of pain during each of four activities (using a visual analogue scale) and assessment of observed disability (walking speed, timed up and go test, and the step test). RESULTS: Therapeutic tape significantly reduced pain on three of the four activities assessed, when compared with the neutral and untaped conditions (P<0.017). The only statistically significant change in observed disability was detected in the step test (P<0.001), in favour of the therapeutic tape. CONCLUSIONS: Therapeutic knee tape is a simple, inexpensive strategy that increases the treatment options for therapists and patients in the conservative management of knee OA. Whilst effective in immediately reducing pain, it does not appear to have a significant immediate impact on observed disability associated with the disease. Therapeutic tape may be used as an adjunct to drug and exercise therapies, potentially augmenting the individual benefits of each. PMID- 12730547 TI - Joint counts in routine practice. PMID- 12730548 TI - Natural killer cells in the synovial fluid of rheumatoid arthritis patients exhibit a CD56bright,CD94bright,CD158negative phenotype. AB - BACKGROUND: Natural killer (NK) cells play an important role in several animal models of autoimmunity by modulating T-cell responses, but it is unclear whether human NK cells have similar functions. METHODS: We characterized the phenotype of NK cells in synovial fluid (SF) and peripheral blood (PB) of patients with rheumatoid arthritis (RA) and in healthy control subjects using flow cytometry and quantitative PCR. RESULTS: The proportions of NK cells in PB and SF of RA patients were not significantly different from those in healthy PB. However, the SF NK cell phenotype was strikingly different, with increased CD94 and CD56 densities and greatly reduced proportions of cells expressing CD158a/b. These cells also had reduced mRNAs coding for CD158a/b and low perforin levels compared with RA PB and healthy PB NK cells. CONCLUSIONS: We identified a novel phenotype of SF NK cells that is of potential significance in RA. Experiments are now under way to determine the function of these SF NK cells and their potential role in RA. PMID- 12730549 TI - Pharmacoutilization and costs of osteoarthritis: changes induced by the introduction of a cyclooxygenase-2 inhibitor into clinical practice. AB - OBJECTIVE: To establish whether the introduction of a cyclooxygenase-2 inhibitor has led to changes in pharmacoutilization in the treatment of osteoarthritis (OA) in clinical practice. METHODS: Administrative and general practice databases were cross-linked to analyse the use of non-steroidal anti-inflammatory drugs (NSAIDs) and gastroprotective agents (GPAs) before and after the introduction of rofecoxib. Costs of treatment and costs of hospitalization for gastrointestinal events were also considered. RESULTS: A total of 3090 patients were evaluated. A significant reduction in the use of GPAs in the rofecoxib group was observed, corresponding to reductions of 64 and 59.7% compared to NSAIDs among patients in incident and prevalent cases respectively. The weighted mean daily cost of therapy with rofecoxib in incident cases was 1.88 euro, 7.4% lower than that of NSAIDs (2.03), and in prevalent cases it was 1.87 euro, 28.1% higher than that of NSAIDs (1.46). Although the rate of hospitalization was similar, there was an additional daily cost per patient of 186.6 for patients being treated with NSAIDs and 21.6 euro for those being treated with rofecoxib. CONCLUSIONS: The cyclooxygenase-2 inhibitor rofecoxib determined substantial changes in the pharmacoutilization and costs of OA. PMID- 12730550 TI - Joint stiffness in a phantom limb: evidence of central nervous system involvement in rheumatoid arthritis. AB - OBJECTIVE: The nature and cause of perceived joint stiffness (PJS), a well established and defining symptom of rheumatoid arthritis (RA), remains unclear. We hypothesized that changes in the central nervous system (CNS) may determine and maintain this subjective experience of stiffness in a limb even after it is amputated. To test this hypothesis, patients with a phantom limb (PL) who had experienced characteristic RA stiffness prior to amputation were systematically investigated. METHODS: Three patients with a current diagnosis of RA and lower limb amputation were investigated to determine the nature and pattern of pain and stiffness in their PL and intact limb. In addition to standard physical examination, pain and stiffness severity was measured using visual analogue scales for both limbs. The duration and timing of stiffness were also recorded for each limb. RESULTS: In all three cases, the pattern of perceived RA stiffness was similar for the intact limb and the PL. All three patients described stiffness in their PL which mirrored that of physical RA joint symptoms in terms of quality, frequency, diurnal variation, location, distribution and response to medication [non-steroidal anti-inflammatory drug (NSAID), corticosteroid, opiate and disease-modifying anti-rheumatic drug (DMARD)]. Unilateral exercise (or attempted exercise) relieved stiffness only in the limb being exercised. CONCLUSION: The extent to which the subjective experience of perceived stiffness could be dissociated from the assumed original peripheral source was strikingly illustrated in RA patients with phantom limbs. We suggest that the PJS characteristic of RA is generated and maintained by secondary plastic changes in the CNS, although causally related to the initial peripheral rheumatoid disease process. PMID- 12730551 TI - Cross-reactivity between anti-cardiolipin, anti-high-density lipoprotein and anti apolipoprotein A-I IgG antibodies in patients with systemic lupus erythematosus and primary antiphospholipid syndrome. AB - OBJECTIVES: Atherosclerosis is an important complication of patients with systemic lupus erythematosus (SLE) and primary antiphospholipid syndrome (APS). One suggested mechanism may be the action of autoantibodies directed against plasma lipoproteins. We studied the presence and patterns of cross-reactivity between antibodies directed against cardiolipin, high-density lipoprotein (HDL) and apolipoprotein A-I (Apo A-I) in patients with SLE and APS. METHODS: Sera from 50 patients (25 SLE and 25 APS) and 10 healthy controls together with three human immunoglobulin G anti-cardiolipin (CL) monoclonal antibodies (IS4, CL1 and CL24) were assessed for the presence of autoantibodies binding cardiolipin, HDL and Apo A-I. Classical inhibition assays were performed to study interference by HDL and Apo A-I in the binding of the human monoclonal antibody to CL. To determine the cross-reactivity patterns between these autoantibodies, sera from 12 patients were incubated on ELISA plates coated with the three different molecules and the captured antibodies were then tested for their activity towards each of the other antigens. RESULTS: All three monoclonals bound to CL. IS4 and CL1 also bound to HDL but only IS4 bound to Apo A-I. Anti-cardiolipin titres were higher in patients with APS than SLE and in healthy controls (P<0.03 and P<0.009 respectively). Titres of antibodies to HDL were higher in patients with SLE and APS than in controls (P<0.009 and P<0.03 respectively). There were no significant differences with respect to the presence of antibodies binding to Apo A-I. In the SLE population, anti-HDL antibody titres correlated with anti-Apo A-I (r=0.563, P<0.004), but not in patients with APS. In the cross-reactive assay, 11/12 (91.7%) of the samples containing isolated anti-CL antibodies reacted to HDL and 2/12 (16.7%) to Apo A-I. Samples containing isolated anti-HDL antibodies also reacted with CL and Apo A-I (7/12 and 3/12 respectively). All samples collected after incubation with Apo A-I bound to HDL and 6/12 (50%) to CL. There were no differences in the cross-reactivity patterns between patients with SLE and APS. CONCLUSIONS: Patients with SLE and APS have antibodies directed against HDL and Apo A-I. A high percentage of these antibodies cross-react with CL, suggesting the presence of different groups of antibodies with different targets. The study of the interaction between the immune response and the lipoprotein components and the correct characterization of the reactivity patterns of autoantibodies may be of relevance in the study of atherosclerosis in patients with SLE and APS. PMID- 12730552 TI - Rheumatologists' opinions on the feasibility of a measurement feedback system in rheumatoid arthritis and the influence of motivation. AB - OBJECTIVE: To assess rheumatologists' opinions about the feasibility of a measurement feedback system in rheumatoid arthritis (RA) and to analyse if motivational aspects play a role in assessing the value of the system and in determining the extent to which it is used. METHODS: A survey sample (n=105) was randomly selected from participants of a measurement feedback system. A survey questionnaire assessed opinions on system outcome, structures and processes, motivation and overall satisfaction. Survey results are given descriptively and groups differing in motivation are compared. RESULTS: The overall response rate was 62%. The system was generally perceived to fulfil its aims, but the effort required to use the system was rated less positive. Rheumatologists had as their motivation either 'science/obligation' or 'individual patient evaluation'. Rheumatologists with the latter motivation were more satisfied with the measurement feedback system, perceived its feasibility as better, and made more use of it. CONCLUSION: Motivation for participating in a measurement feedback system has a significant impact on overall satisfaction with the system and the use of the system. Influencing motivation and reduction of the amount of effort required to use the system might increase overall acceptance. PMID- 12730553 TI - Adenovirus-mediated gene transfer of fibroblast growth factor-2 increases BrdU positive cells after forebrain ischemia in gerbils. AB - BACKGROUND AND PURPOSE: Progenitor cells continue to generate neurons in the adult mammalian brain, and cerebral ischemia induces neurogenesis. We examined the efficacy of the intraventricular injection of a recombinant adenovirus expressing fibroblast growth factor-2 (FGF-2) (AxCAMAssbFGF) on neurogenesis in both normal and ischemic brains. METHODS: We used a gerbil model of transient global ischemia and counted the number of BrdU-positive cells after injection of AxCAMAssbFGF into the brain with or without ischemia. RESULTS: Intraventricular AxCAMAssbFGF produced robust FGF-2 protein increases in diverse regions of the brain and markedly increased FGF-2 concentrations in cerebrospinal fluid 2 days after administration and evoked significant proliferation of BrdU-positive cells not only in the subventricular zone and dentate gyrus of the hippocampus but also in the cerebral cortex, and some BrdU-positive cells differentiated into neurons. Continuous intraventricular infusion of FGF-2 protein increased FGF-2 concentration in cerebrospinal fluid but not in brain tissues and produced BrdU positive cell proliferation only in the subventricular zone of the lateral ventricle. CONCLUSIONS: Adenovirally mediated transfer of the FGF-2 gene promoted progenitor cell proliferation more efficiently in widespread regions of the brain after transient global ischemia than continuous intraventricular infusion of FGF 2 protein. PMID- 12730555 TI - Expanded modes of tissue plasminogen activator delivery in a comprehensive stroke center increases regional acute stroke interventions. AB - BACKGROUND AND PURPOSE: We sought to evaluate whether a comprehensive stroke center could work with regional hospitals to increase the use of tissue plasminogen activator (tPA) in acute stroke. METHODS: In 30 months, 142 patients seen at the Mid America Brain and Stroke Institute received tPA. Site of presentation, protocol selection, and outcomes were analyzed. RESULTS: We found that 18.2% (142 of 781) of all ischemic strokes received tPA. Of those, 70% (99 of 142) were transferred from hospitals within 100 miles of Kansas City (Mo). Mortality rate was 12.7% (18 of 142). Symptomatic hemorrhage rate was 9.2%. CONCLUSIONS: A comprehensive stroke center can serve as a hub for a regional network and increase the number of stroke interventions with acceptable outcomes. PMID- 12730554 TI - Neuroprotection in transient focal cerebral ischemia by combination drug therapy and mild hypothermia: comparison with customary therapeutic regimen. AB - BACKGROUND AND PURPOSE: A combined therapeutic approach has been advocated repeatedly for treatment of focal cerebral ischemia. A clinical example of combined therapy is administration of nimodipine, mannitol, dexamethasone, and barbiturates during temporary occlusion of a cerebral artery in neurovascular surgery. We have recently demonstrated outstanding neuroprotective properties of a combination therapy with magnesium (calcium antagonist and glutamate antagonist), tirilazad (antioxidant), and mild hypothermia (MTH). In this study we compared this treatment strategy with the customary treatment options in a rat model of transient focal cerebral ischemia. METHODS: Sprague-Dawley rats (n=120) were subjected to 90 minutes of middle cerebral artery occlusion by an intraluminal filament (n=10 per group). In experiment 1, the customary treatment options (nimodipine, mannitol, dexamethasone, methohexital) were evaluated as monotherapy and in combination. In experiment 2, the customary and the new combination therapy (MTH) were compared. Mild hypothermia (33 degrees C) was maintained for 2 hours. Neurological examinations were performed daily. Infarct size was assessed histologically after 7 days. RESULTS: In experiment 1, infarct volume was attenuated by 34% at maximum, with mannitol and methohexital being the most effective drugs given as monotherapy. In experiment 2, combined administration of the customary treatment options had no additive effect (infarct volume -36%). Combination therapy with MTH reduced total infarction by 73% and almost completely abolished cortical infarction (-91%). None of the animals of this group had any residual neurological deficit at the end of the observation period (P<0.05 versus all other groups). CONCLUSIONS: The efficacy of drugs (monotherapy or in combination) most commonly used for neuroprotection during neurovascular surgery is limited. The newly proposed combination therapy (magnesium, tirilazad, and mild hypothermia), which is based on pathophysiological considerations, seems to be a promising alternative for neuroprotection in cerebrovascular surgery. PMID- 12730556 TI - Pioglitazone improves insulin sensitivity among nondiabetic patients with a recent transient ischemic attack or ischemic stroke. AB - BACKGROUND AND PURPOSE: The aim of this study was to determine the effectiveness of pioglitazone compared with placebo for improving insulin sensitivity among nondiabetic patients with a recent transient ischemic attack (TIA) or nondisabling ischemic stroke and impaired insulin sensitivity. METHODS: Eligible subjects were men and women >45 years of age who had no history of diabetes, fasting glucose <7.0 mmol/L, and impaired insulin sensitivity according to an index calculated from insulin and glucose blood levels obtained during an oral glucose tolerance test. Eligible subjects were randomized to pioglitazone 45 mg/d or placebo. After 3 months of therapy, the glucose tolerance test was repeated. RESULTS: Between July 2000 and June 2001, we performed oral glucose tolerance tests on 75 patients with no history of diabetes, among whom 36 (50%) were found to have impaired insulin sensitivity and fasting glucose <7.0 mmol/L. Among these 36, 20 consented to the trial. Patients assigned to pioglitazone (n=10) and placebo (n=10) were similar in insulin sensitivity, age, obesity, and index event (stroke compared with TIA), but patients assigned to pioglitazone were less likely to be male (4 compared with 9). The mean proportional increase in insulin sensitivity was 62% among patients assigned to pioglitazone compared with a -1% decline among patients assigned to placebo (P=0.0006). Mean C-reactive protein concentration declined from 0.30 to 0.20 mg/L among patients assigned to pioglitazone and increased from 0.41 to 0.45 mg/L among patients assigned to placebo (P=0.06 for comparison of mean change). CONCLUSIONS: Pioglitazone is effective for improving insulin sensitivity among patients with recent TIA or stroke and impaired insulin sensitivity. PMID- 12730557 TI - Effect of hypertonic saline on cerebral blood flow in poor-grade patients with subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: The goal of this study was to examine the effects of hypertonic saline on cerebral blood flow (CBF) in poor-grade patients with subarachnoid hemorrhage. METHODS: We administered 23.5% hypertonic saline (2 mL/kg IV) 1 time to 10 patients, 2 times to 7 patients, and 3 times to 1 patient. All patients had transcranial Doppler (TCD), intracranial pressure (ICP) monitoring, and analysis of serum sodium and osmolality; 6 had xenon CT (XeCT). Data were used to characterize the changes in CBF, cerebral vascular resistance (CVR), ICP, cerebral perfusion pressure (CPP), and potential rheological mechanisms of action. RESULTS: In the first treatment episode, CPP increased 26.8% (P=0.0003, at 28.3 minutes) from a rise in mean arterial blood pressure (ABP) of 10.5% (P=0.02, at 22.2 minutes) and a fall in ICP (-74.7%, P=0.002, at 60.0 minutes). Flow velocity (FV) of the middle cerebral artery increased 70.8% (P=0.00005, at 20.0 minutes), resulting in a corresponding fall in estimated CVR (-26.6%, P=0.01, at 16.3 minutes). The half-lives of effects on ABP, CPP, ICP, FV, and estimated CVR were 20.0, 53.6, 139.1, 42.7, and 27.1 minutes, respectively. In the second treatment episode, all these parameters had the same response except estimated CVR, which did not reach statistical significance. XeCT confirmed the increase in CBF (22.9%, P=0.02) without regional differences. A fall in CBF after hypertonic saline was identified in only a single region of interest in a patient in whom baseline flow was low but not infarcted. Serum sodium rose by 11.4 and 8.8 mmol/L, and osmolality rose by 26.7 and 16.3 mosm/L in the first and second treatment episodes, respectively. Hemoglobin decreased by 0.7 and 0.6 g/L and hematocrit decreased by 1.9% and 2.4% in the first and second treatment episodes, respectively. CONCLUSIONS: We found that 23.5% hypertonic saline increases CBF in poor-grade patients with subarachnoid hemorrhage. These effects are associated with improved indexes of blood rheology. Potential therapeutic benefits are discussed. PMID- 12730558 TI - Comparison of P2 receptor subtypes producing dilation in rat intracerebral arterioles. AB - BACKGROUND AND PURPOSE: P2 receptors are important regulators of cerebrovascular tone. However, there is functional heterogeneity of P2Y receptors along the vascular tree, and the functionality of P2Y receptors in small arterioles has not been studied in detail. We investigated the effects of activating P2Y1 and P2Y2 receptors and their underlying dilator mechanisms in rat intracerebral arterioles. METHODS: We used computer-aided videomicroscopy to measure diameter responses from isolated and pressurized rat penetrating arterioles (39.9+/-1.2 microm) to the natural P2 receptor agonist ATP in addition to ADP-beta-S (P2Y1 selective) and ATP-gamma-S (P2Y2-selective) and inhibitors of signaling pathways. RESULTS: Extraluminal application of ATP-gamma-S and ADP-beta-S initiated a biphasic response (initial constriction followed by the secondary dilation) similar to ATP-induced responses. Pyridoxal phosphate-6-azophenyl-2',4' disulphonic acid (0.1 mmol/L; a P2Y1 receptor antagonist) blocked ADP-beta-S- but not ATP-gamma-S-induced dilation and affected the ATP-mediated dilation at low concentrations. Nomega-Monomethyl-l-arginine partially inhibited the dilation of ATP and ADP-beta-S but not ATP-gamma-S. High K+ saline suppressed the dilation of all agonists. Indomethacin had no effect. CONCLUSIONS: Both P2Y1 and P2Y2 receptors are functionally present in cerebral arterioles. ATP stimulates P2Y1 receptors at low concentrations, while high concentrations of ATP activate P2Y2 in addition to P2Y1 receptors. Nitric oxide is involved in P2Y1 but not P2Y2 receptor activation. Potassium channels play an important role in the regulation of P2Y receptor-mediated dilation. PMID- 12730560 TI - Thrombolytic therapy with recombinant tissue plasminogen activator for acute ischemic stroke: where do we go from here? A cumulative meta-analysis. AB - BACKGROUND AND PURPOSE: Recombinant tissue plasminogen activator (rtPA; Actilyse) is not as widely used in clinical practice as it could be. Have new data since 1995 strengthened the evidence sufficiently to justify more widespread use of rtPA? METHODS: We performed a sequential year-to-year cumulative meta-analysis of randomized controlled trials of rtPA in acute ischemic stroke. RESULTS: Although the amount of data has doubled since 1995, effect estimates for key outcomes remain imprecise, and significant between-trial heterogeneity persists. In the most recent analysis, rtPA up to 6 hours after stroke yielded 55 fewer dead or dependent people per 1000 treated (95% CI, 18 to 92) despite some risk (nonsignificant excess of 19 deaths per 1000 patients treated; 95% CI, 6 fewer to 48 more). Severity of stroke, patient age, and aspirin use were possible sources of heterogeneity. CONCLUSIONS: Despite doubling of the data since 1995, the magnitude of risks and benefits with rtPA remains imprecise. This gap in knowledge may be hindering clinical use of rtPA and can be filled only by new trials designed to address these specific issues. PMID- 12730561 TI - Lessons from SARS. PMID- 12730559 TI - Oral anticoagulation in patients after cerebral ischemia of arterial origin and risk of intracranial hemorrhage. AB - BACKGROUND AND PURPOSE: In the recently published Warfarin Aspirin Recurrent Stroke Study (WARSS), a low-intensity anticoagulation regimen was used because of safety concerns. Such concerns are corroborated by the results of the Stroke Prevention in Reversible Ischemia Trial (SPIRIT), which was stopped early because of a high incidence of intracranial hemorrhage with a target international normalized ratio (INR) of 3.0 to 4.5. In the ongoing European/Australasian Stroke Prevention in Reversible Ischaemia Trial (ESPRIT), an intermediate anticoagulation regimen (INR 2.0 to 3.0) is used. METHODS: We performed an interim analysis of the incidence of intracranial hemorrhage in ESPRIT. RESULTS: Thus far the overall rate of intracranial hemorrhage is 0.31% (95% CI, 0.18% to 0.52%) per year and 1.21% if all of these were in the anticoagulation group. CONCLUSIONS: We conclude that anticoagulation with achieved INR of 2.0 to 3.0 is reasonably safe in patients with cerebral ischemia of arterial origin. PMID- 12730563 TI - Infectious diseases. Hungry for details, scientists zoom in on SARS genomes. PMID- 12730562 TI - Infectious diseases. Battling SARS on the frontlines. PMID- 12730564 TI - Infectious diseases. WHO wants 21st-century reporting regs. PMID- 12730566 TI - Planetary exploration. NASA bails out of French-led Mars mission. PMID- 12730565 TI - Infectious diseases. Avian flu outbreak sets off alarm bells. PMID- 12730568 TI - Neuroscience. Old neurons revisit their youth. PMID- 12730567 TI - Stem cells. Oocytes spontaneously generated. PMID- 12730569 TI - Developmental biology. Purified signaling protein stimulates stem cell proliferation. PMID- 12730570 TI - Archaeology. Tortoise pace for the evolution of Chinese writing? PMID- 12730571 TI - Science philanthropy. Gates grows UW's genome program. PMID- 12730573 TI - Planetary science. Moon maintains its mysteries. PMID- 12730572 TI - Planetary science. The new race to the Moon. PMID- 12730574 TI - Biotechnology. Hatching the golden egg: a new way to make drugs. PMID- 12730575 TI - Cosmology. With its ingredients MAPped, universe's recipe beckons. PMID- 12730576 TI - Editorial retraction (I). PMID- 12730577 TI - Editorial retraction (II). PMID- 12730578 TI - Protecting Iraqi antiquities. PMID- 12730579 TI - Science publishing and security concerns. PMID- 12730580 TI - Ethnic differences and disease phenotypes. PMID- 12730581 TI - Studying traditional Chinese medicine. PMID- 12730582 TI - Comment on "Grasping primate origins". PMID- 12730585 TI - Atmospheric science. On sprites and their exotic kin. PMID- 12730584 TI - Perceptions of science. Prebiotic soup--revisiting the Miller experiment. AB - Also see the archival list of the Essays on Science and Society. PERCEPTIONS IN SCIENCE: Prebiotic Soup--Revisiting the Miller Experiment Jeffrey L. Bada and Antonio Lazcano PMID- 12730586 TI - Chemistry. The beauty of symmetry. PMID- 12730587 TI - Signal transduction. Autoinhibition control. PMID- 12730588 TI - Astronomy. Black holes at the cosmic dawn. PMID- 12730589 TI - Molecular biology. A place to die, a place to sleep. PMID- 12730590 TI - Structural biology. Changing partners. PMID- 12730591 TI - Computational biology. Counting on the neuron. PMID- 12730592 TI - Assessing the impact of the green revolution, 1960 to 2000. AB - We summarize the findings of a recently completed study of the productivity impacts of international crop genetic improvement research in developing countries. Over the period 1960 to 2000, international agricultural research centers, in collaboration with national research programs, contributed to the development of "modern varieties" for many crops. These varieties have contributed to large increases in crop production. Productivity gains, however, have been uneven across crops and regions. Consumers generally benefited from declines in food prices. Farmers benefited only where cost reductions exceeded price reductions. PMID- 12730593 TI - Chimeric nucleases stimulate gene targeting in human cells. PMID- 12730595 TI - Supramolecular self-assembly of lipid derivatives on carbon nanotubes. AB - Images of the assembly of surfactants and synthetic lipids on the surface of carbon nanotubes were obtained by transmission electron microscopy. Above the critical micellar concentration, sodium dodecyl sulfate (SDS) forms supramolecular structures made of rolled-up half-cylinders on the nanotube surface. Depending on the symmetry and the diameter of the carbon nanotube, we observed rings, helices, or double helices. Similar self-assemblies were also obtained with several synthetic single-chain lipids designed for the immobilization of histidine-tagged proteins. At the nanotube-water interface, permanent assemblies were produced from mixed micelles of SDS and different water insoluble double-chain lipids after dialysis of the surfactant. Such arrangements could be further exploited for the development of new biosensors and bioelectronic nanomaterials. PMID- 12730594 TI - Enhancing gene targeting with designed zinc finger nucleases. PMID- 12730596 TI - Interpenetrating As20 fullerene and Ni12 icosahedra in the onion-skin [As@Ni12@As20]3- ion. AB - The [As@Ni12@As20]3- ion was prepared from As7(3-) and Ni(COD)2 in ethylenediamine solutions and isolated as the Bu4P+ salt (As, arsenic; Ni, nickel; COD, cyclooctadiene; Bu, butyl; P, phosphorus). The anion contains an icosahedral [Ni12(mu12-As)]3- fragment that resides at the center of an As20 dodecahedral (fullerene) cage to give an onion-skin-like [As@Ni12@As20]3- cluster with Ih point symmetry. The icosahedron and pentagonal dodecahedron are reciprocal platonic solids, and the 32 surface atoms form a dimpled geodesic sphere composed of 60 triangular faces. In the gas phase, the [As@Ni12@As20]3- ion sequentially loses all 21 As atoms to form a series of Ni12As(21-x) clusters where 0 400 seconds in 29.0%, 29.0% and 42.1%, respectively. In-hospital clinical events occurred in 12 patients (9.2%) who met any one of the following endpoints: death (0.8%), limb loss (1.5%), major bleeding (4.6%), emergent need for repeat revascularization of the same vessel (7.6%), embolic stroke (0.0%) and vascular complications (1.5%). The best model associated with salvage revascularization included cigarette smoking within the past year, recent onset of claudication and PTA treatment below the knee. Increased dosages of heparin (U/kg) were associated with a trend toward higher rates of complications. A significant number of patients have in-hospital major complications following PTA procedures using unfractionated heparin as the primary anticoagulant. Current ongoing registries are evaluating the feasibility of direct thrombin inhibitors bivalirudin instead of heparin as a primary anticoagulant during PTA. PMID- 12730631 TI - Heparin in peripheral vascular intervention--time for a change? PMID- 12730632 TI - Clinical effectiveness of the Prostar XL suture-mediated percutaneous vascular closure device following PCI: results of the Perclose AcceleRated Ambulation and DISchargE (PARADISE) Trial. AB - BACKGROUND: The Prostar XL is a hemostasis device designed to percutaneously close the arterial puncture site with two nonabsorbable sutures. The Early Discharge and Economical effectiveness study for New hemostasis device (EDEN) trial, which was performed in Japan, showed that the Prostar XL device shortened time to hemostasis, time to ambulation and the average length of hospital stay, and decreased local vascular complications. However, the ooze of blood from the puncture site incision requires relatively longer time to ambulation. OBJECTIVES: The Perclose AcceleRated Ambulation and DISchargE (PARADISE) trial was performed to assess the effectiveness of the Prostar XL device on patient comfort, shortened time to ambulation and discharge from hospital by minimizing the oozing problem. METHODS: This multicenter, non-randomized registry was designed to compare times to hemostasis, ambulation and discharge, as well as the incidence of peripheral vascular complications in Prostar XL patients with the conventional manual compression group in the EDEN trial. Hydrochloride lidocaine containing 1% epinephrine was used as local anesthesia to reduce blood oozing. RESULTS: Between September 1999 and March 2000, a total of 109 patients were enrolled in this study. The closure device was successfully placed in 108 of 109 attempts. Time to hemostasis was significantly less than with the conventional manual compression group in the EDEN trial. Times to ambulation and discharge were significantly less than with the Prostar XL and conventional manual compression groups in the EDEN trial. CONCLUSION: We conclude that the Prostar XL device plus hydrochloride lidocaine containing 1% epinephrine as local anesthesia appears to be a safe and effective method to achieve hemostasis, and to improve times to ambulation and discharge after interventional procedures. PMID- 12730633 TI - Simultaneous multisite endocardial mapping of sustained and non-sustained atrial fibrillation in humans. AB - The aim of this study was to investigate the differences between sustained and non-sustained forms of human atrial fibrillation (AF) using multielectrode endocardial recordings. Methods. Sixty-four pole basket catheters were deployed in the right atrium (RA) of 3 groups of patients: 1) patients with persistent AF (> 48 hours); 2) induced sustained AF (> 15 minutes); and 3) induced non sustained AF (< 15 minutes). Beat to beat AF intervals (FF) were evaluated for each bipole. On the basis of signal characteristics and direction of wavefront propagation, the degree of spatial and temporal organization of AF was assessed. Results. Persistent AF showed the shortest FF intervals (161 ms) and lowest overall degree of AF organization, induced non-sustained AF the longest FF intervals (192 ms) and highest degree of organization. FF intervals of induced sustained AF were only slightly longer (169 ms) compared to persistent AF. Within each AF group, the lateral wall showed the highest degree of organization, the septal region the lowest. Conclusion. In humans, FF interval and overall degree of AF organization were found to increase significantly from sustained to non sustained AF. Persistent and induced sustained AF, however, only slightly differed in these parameters. PMID- 12730634 TI - Atrial fibrillation: don't become a basket case. PMID- 12730635 TI - Platelets and antiplatelet therapy in patients with diabetes mellitus. AB - Atherosclerotic heart disease is the leading cause of death in patients with diabetes mellitus. Platelets play a major role in the clinical manifestations of ischemic heart disease. Diabetic patients have hyperreactive platelets with exaggerated adhesion, aggregation and thrombin generation. Antiplatelet agents, including aspirin, clopidogrel, and glycoprotein IIb/IIIa inhibitors, have shown significant efficacy in reducing recurrent ischemic events in patients with diabetes. Treatment with glycoprotein IIb/IIIa inhibitors during percutaneous coronary intervention reduces mortality in diabetic patients. PMID- 12730636 TI - A rare combination of LMCA fistula to left superior vena cava with drainage into the coronary sinus in a 74-year-old woman. AB - Coronary artery fistulae are rare anomalies in which a communication is present between a coronary artery and either a cardiac chamber or another vascular structure, such as the pulmonary artery or coronary sinus. Most fistulae are congenital in origin, but they can also occur as a result of chest trauma or endocardial biopsy. The presence of a fistula is a rare occurrence in itself, but the different origins and drainage sites of fistulae are what make them even more rare. We report the case of a 74-year-old woman with a history of progressive congestive heart failure who was found to have a large left main coronary fistula draining into a persistent left superior vena cava to a markedly dilated and aneurysmal coronary sinus. PMID- 12730637 TI - Delayed development of a giant coronary pseudoaneurysm after stent placement for chronic total occlusion. AB - A 42-year-old man developed a giant pseudoaneurysm in a stented coronary segment despite excellent results after stent placement for chronic total occlusion. The aneurysm was treated successfully with the deployment of a covered stent. Delayed development of a giant pseudoaneurysm after stent implantation for chronic total occlusion is an uncommon complication. PMID- 12730638 TI - Management of iliac stent movement complicating peripheral vascular intervention: A rescue technique when stent deployment malfunctions. AB - Percutaneous transluminal peripheral angioplasty (PTA), with and without stenting, has become an effective, accepted, and safe approach for treating intermittent claudication and critical leg ischemia in selected patients. Percutaneous peripheral interventions are subject to complications that require unique solutions when compared to coronary interventions. We report a case of common iliac artery stenting complicated by stent migration and describe an approach to the management of this problem. PMID- 12730639 TI - Diagnostic importance of angiographic visualization of distal vessel in elucidating abrupt vessel closure. AB - Acute vessel closure complicates 0.5 9.3% of percutaneous coronary interventional procedures. This case series demonstrates the potential value of distal vessel angiographic visualization during abrupt vessel closure, which allows rapid determination of the underlying mechanism and the subsequent appropriate therapy. PMID- 12730640 TI - Refractory vasomotor angina in subclinical hyperthyroidism demonstrating focal and segmental coronary vasoconstriction. AB - We describe a case of refractory vasomotor angina in the setting of unrecognized subclinical hyperthyroidism. Despite aggressive medical therapy, frequent rest angina recurred until diagnosis and treatment of Grave s Disease, which presented asymptomatically. Both spontaneous focal vasospasm and inducible segmental coronary vasoconstriction were demonstrated during invasive provocative testing. Subclinical hyperthyroidism should be considered in the differential diagnosis when treating refractory vasomotor angina, especially in women over 50 years old, a population at particular risk for undiagnosed thyroid disease. PMID- 12730641 TI - The use of the X-Sizer transluminal extraction catheter as an adjunct to stenting of occluded saphenous vein grafts. PMID- 12730642 TI - Adjunctive use of cutting balloon after rotational atherectomy in a young adult with probable kawasaki disease. PMID- 12730643 TI - [Clinical concern for antibiotic resistance] AB - Antibiotic resistance represents such a serious problem in the field of infectious diseases as to constitute as a threat for a post-antibiotic era. Currently, it contributes to increase treatment failures, morbidity and mortality rates, and health costs. Since 1993, the international scientific community has identified five different priorities in the struggle against antibiotic resistance which are represented by the following bacterial species: Mycobacterium tuberculosis, Streptococcus pneumoniae, Staphylococcus spp, Enterococcus spp and Neisseria gonorrhoeae. At present three different approaches have been identified to overcome this phenomenon: identification of mechanisms of bacterial resistance; epidemiological surveys; synthesis of new molecules active against multiresistant microrganisms.Besides the above mentioned strategies, preventive interventions should also be carried out mainly through the optimisation of antibiotic prophylaxis, optimisation of choice and duration of therapy, improvement of antibiotic prescriptions by means of educational and administrative programs, monitoring and feedback of bacterial resistance and finally by the definition and diffusion of guidelines for therapy. PMID- 12730644 TI - [Association between chronic hepatitis C virus infection and cryoglobulinemia] AB - Hepatitis C virus infection (HCV) causes both acute and chronic liver disease and can be also associated with cryoglobulinemia (SC). SC is a systemic vasculitic disease, typically characterized by lower extremity purpure, arthralgias and fatigue and by circulating immune complexes which precipitate at low temperatures. We examined the prevalence of SC in a prospective study of 84 patients with chronic HCV hepatitis. Cryoglobulinemia was detected in 44 patients (53.4%) and was associated with the severity of liver damage and the duration of the disease. The analysis of HCV genotypes demonstrated a prevalence of 1 b. The amount of cryoglobulinemia was low in all the patients with SC and only 20% showed a clinical syndrome. PMID- 12730645 TI - [Glucagon test for Doppler sonography measurement of portal flow in chronic HCV infections] AB - Purpose - To evaluate whether Doppler sonography measurement of portal flow velocity (PFV) after glucagon injection can be useful in assessing the severity of liver damage in chronic HCV infection. Methods - Forty-five patients (32 males and 13 females; mean age 54.1 14.8 years) with biochemical (raised serum ALT levels), virological (positive serum HCV RNA test) and histological (liver biopsy) evidence of chronic HCV infection were included in the study. According to hepatitis staging (degree of liver fibrosis), as assessed by Knodell histological activity index, patients were divided into three groups: group 1 (n.=17), with no or mild fibrosis (fibrosis score: 0-1); group 2 (n.=11), with severe fibrosis (score: 3); and group 3 (n.=17), with liver cirrhosis (score: 4). For sonographic measurements of PFV, a Doppler ultrasound multi-purpose equipment and a convex 3.5 MHz probe were used. All patients were examined after an 8-hour fast, in supine position, 10 min before (baseline), as well as 5 and 10 min after, intravenous administration of 1 mg of glucagon chloride (Novo Nordisk). Statistical analysis was performed by ANOVA test, Bonferroni t test and Spearman rank correlation test. Results - No significant differences were found in mean basal PFV of group 1 (19.4 2.4 cm/sec), group 2 (20.1 3.6 cm/sec) and group 3 (17.5 3.7 cm/sec) (p > 0.05). Five minutes after glucagon injection, all the three groups showed a significant increase in PFV (25.6 4.8,23.7 4.0 and 19.5 5.0 cm/sec, respectively; p < 0.05 vs baseline). The peak increase in PFV after glucagon injection was significantly higher in group 1 (7.9 3.7 cm/sec; 40.7% of basal value) than in group 2 (4.5 3.9 cm/sec; 22.4%) (p < 0.05) and in group 3 (2.7+2.3 cm/sec; 15.4%) (p < 0.05). A significant (p< 0.001) inverse correlation was also found between the patients fibrosis scores and peak increments of PFV induced by glucagon. Conclusions - In some patients with chronic HCV infection, Doppler sonography measurement of PFV after glucagon injection can be useful, in combination with other non invasive ultrasound investigations, both in staging of liver disease and in monitoring the progression of liver histological damage. PMID- 12730646 TI - [Attempted suicide during interferon therapy: a case report and literature review] AB - Occasionally, in the medical literature, attempted or successful suicides during interferon therapy are reported. We report a case of a 32-year-old man who developed a psychotic syndrome with attempted suicide at the third month of treatment. The cessation of interferon and an appropriate psychiatric therapy determined a complete remission of symptoms. A medical literature review, through several computerized searches, showed a lack of research into the subject. The main conclusion of our article emphasizes the importance of recognizing the early neuropsychiatric symptoms, that are reversible at the cessation of treatment. PMID- 12730647 TI - [Neurologic signs in bacterial endocarditis: an unusual case report] AB - We describe the case of a 35-year-old woman, hospitalised due to a fever lasting two days, with signs and symptoms of cerebro-spinal meningitis. The subsequent occurrence of a thromboembolic event with stroke suggests that the neurological symptoms were secondary to a centre of infective endocarditis (instrumentally proved) in the absence of further outstanding symptoms of endocarditis. This event is very unusual during the course of endocardic disease. PMID- 12730649 TI - [Il cholera morbus nel Comune di Bologna nel 1886] AB - Not available PMID- 12730648 TI - [CD30 (ki-1) antigen expression in acute infectious mononucleosis] AB - The activation antigen CD30 (Ki-1) which is expressed by the lymphocytes T and B is related to the "nerve growth factor" (NFGR) like the receptor of the "tumor necrosis factor" (TNF) and is expressed in the group of large-cell anaplastic lymphoma (ACLC) and Hodgkin's lymphoma and in non-Hodgkin's lymphoma and within the T cells of infectious Mononucleosis. We describe a case of a 18 year-old who contracted Acute Infectious Mononucleosis showing lateral cervical lympho adenopathy. The histologic examination was suggestive for a large-cell anaplastic lymphoma with the antigen CD30, expressed by severa atypical immunoblasts. The remarkable immunoblastic proliferation with Reed-Sternberg-like cells, in milieu polymorphous inflammatory cells present in the course of Infectious Mononucleosis, can simulate large cell anaplastic lymphoma, as well as Hodgkins disease and non-Hodgkins lymphoma. The diagnosis for Infectious Mononucleosis is usually clinical and serological but in cases of cervical lymphnode biopsy a differential diagnosis must be done with positive lymphoma CD30. The criteria to differentiate histologically the Infectious Mononucleosis with large cell anaplastic lymphoma and non-Hodgkin s lymphoma are discussed. Finally, the relationship between Infectious Mononucleosis and lymphoproliferative disease Epstein-Barr virus related is examined. PMID- 12730650 TI - Orofacial neurogenic pain and maxillofacial ischemic osteonecrosis. A review. AB - Cavitary alveolar osteopathy was described as an oral disorder of infectious origin characterized by the presence of osteopathic alveolar cavity lesions of significant size though radiologically undetectable and secondary to dental extractions for chronic infectious processes of the alveolar bone of the jaws. Such cavitary alveolar osteopathy has been implicated as a common cause in the origin of idiopathic trigeminal neuralgia and atypical facial pain. The concept of cavitary alveolar osteopathy caused by ischemic necrosis of alveolar bone was introduced in 1992. Recent coagulation studies have reported ischemic alterations in alveolar bone marrow as a cause of cavitation; following tooth extraction, maxillary osteonecrosis could result from thrombosis with or without hyperfibrinolysis, which in turn would lead to obstruction of the vascular spaces -- thereby compromising regional blood flow. PMID- 12730652 TI - Dental management of the complications of radio and chemotherapy in oral cancer. AB - The most common malignancy of the oral cavity is epidermoid or squamous cell carcinoma, which accounts for approximately 5% of all neoplasms. Unfortunately, the great majority of these tumors are diagnosed in stages which require surgery with radio- and chemotherapy. Radiotherapy constitutes an important option in the treatment of oral tumors, and can be applied either alone or in combination with surgery and chemotherapy. The latter has no precisely defined role in the treatment of squamous cell carcinomas, and is usually used as a coadjuvant therapy or for palliative purposes. Since these treatments affect not only the malignant cells but also the healthy tissues of the patient, side effects usually develop during and after treatment, in the form of oral lesions and systemic alterations. Examples include mucositis, xerostomia, immune suppression, and viral and fungal infections, among other problems. The present study offers a management protocol for the oncological patient before, during and after radio- and chemotherapy. In addition, emphasis is placed on the important role of the dental professional in the prevention and treatment of the main oral complications, proposing dental management guidelines which are applicable in the general clinical context. PMID- 12730651 TI - Radiographic features of osseous hemangioma in the maxillo-facial region. Bibliographic review and case report. AB - Osseous hemangioma in the maxillofacial region (OHMF) was first reported in the literature as a clinical entity in the middle of last century. Several fatal cases of accidental hemorrhage following tooth extraction were attributed to OHMF. The discovery of X-rays and image diagnosis should have facilitated identification of the lesion and contributed to establishing differential diagnosis with other entities. However, our review of case-reports suggests that this lesion has been misdiagnosed and mistaken for other neoplastic and non neoplastic hemorrhagic bone lesions. The aim of this study was to analyze clinical and radiographic data obtained from our review in order to determine the diagnostic criteria for OHMF over the last twenty years. Our case study showed that OHMF occurred at a mean age of 27.3 years, and appeared at a later age in women (30.4 years). OHMF was more frequent in women with a female to male ratio of 1.6:1, although a higher ratio has been reported in the literature. The site of predilection was the premolar and molar region of the mandible (65%). Clinical diagnosis of hemangioma was performed in 7 cases only (35%); diagnosis of the remaining cases was less accurate. The most frequent radiographic feature was mixed radiodensity (70%) and radiolucency (30%). A honeycombed appearance predominated (35%). In addition, 8 cases of OHMF involved the teeth which exhibited resorption and displacement. The present study shows that image diagnosis of OHMF must be based on the critical application of increasingly complex techniques in order to define the lesion. PMID- 12730653 TI - Effect of Chitosan and Sodium Alginate on the adherence of autochthonous C. Albicans to oral epithelial cells (in vitro). AB - The aim of the present study was to evaluate the effect of Heavy Molecular Weight Chitosan (HMWCh) and Sodium Alginate (NaAl) on fungal adherence. C albicans was identified and isolated from non-stimulated saliva extracted from male and female healthy adults. The minimum inhibitory concentration (MIC) was determined for each of the biopolymers. MIC values were 0.25 % (W/V) for HMWCh and 0.10 % (W/V) for NaAl. Fungal cell hydrophobicity was evaluated against xylene in the presence of HMWCh. Statistically significant differences between the control (without HMWCh) and the different HMWCh concentrations in fungal suspension were observed (P< 0.05). The fact that HMWCh and NaAl impaired fungal adherence to buccal epithelial cells (BEC) as compared to control revealed that polymers inhibit Candida albicans adherence to BEC (HMWCh and NaAl: P= 0.00001), NaAl being more effective than HMWCh (P = 0.00001). HMWCh dettached and aggregated C. albicans, including the fungi and BEC in the mesh. NaAl inhibited adherence, aggregated and entrapped the fungi in the mesh, excluding BEC. We may conclude that both biopolymers are effective. However, NaAl is a stronger inhibitor of adherence. Thus, in combination or alone, these biopolymers could be used in the treatment of oral candidosis. PMID- 12730654 TI - Appearance and culture: oral pathology associated with certain "fashions" (tattoos, piercings, etc.). AB - Humans are characterized by a compulsive tendency to distinguish themselves from the rest: differences in clothes, hairstyle or "decorative" details are used to this effect, based on highly diverse criteria. Such differentiating practices may be aimed at identification with a certain ideological group, for example, or with a concrete "fashion", and involve the use of jewelry, clothes, unusual attire, hairstyles, mutilations, etc. In this context, the present review addresses certain aspects of mutilation practices from both the general and specifically dental perspectives. Mutations imply permanent or lasting sectioning or lesions of a part of the body, and comprise skeletal deforming, dental mutilations, circumcision, ablation of the clitoris, scarification, tattoos, and perforations (particularly of the soft tissues). In this sense, tattoos and perforations or piercings are popular -- particularly among adolescents. This trend may be interpreted as a form of communication, identity expression, or as a type of body cult (i.e., so-called "body art"). Such mutilating practices reflect different motivations including fashion, rebelliousness, differentiation, sexual motives, the remembering of events, physical sensations, and ethnic or tribal influences. However, these practices can cause complications such as infections, laceration and soft and hard tissue damage, hypersensitivity reactions and other alterations of variable severity. Under these premises, questions are raised concerning the competence of those who perform these mutilations, the preventive measures adopted, and the legal conditions under which tattoos and piercings are made in our society. PMID- 12730655 TI - Management of patients with adrenocortical insufficiency in the dental clinic. AB - Patients with systemic disorders are often seen in dental practice. If not adequately evaluated, the existence of such alterations may lead to undesirable clinical situations. This is the case of patients with adrenocortical insufficiency. The causes underlying such insufficiency require a distinction between primary adrenal insufficiency or Addison's disease, involving an intrinsic alteration of the adrenal gland cortex, and secondary adrenal failure attributable to hypophyseal or hypothalamic pathology. Another frequent situation involves patients subjected to high-dose steroid therapy -- which induces a reduction in ACTH (adrenocorticotropic hormone) secretion with secondary atrophy of the adrenal cortex. In this context, sudden interruption of corticoid therapy gives rise to acute adrenocortical insufficiency, particularly if the patient is subjected to some form of stress. Corticotherapy is provided in many disorders such as autoimmune connective tissue diseases, different types of cancer, blood dyscrasias and transplant patients, among others. This makes it likely for dentists to occasionally see patients who are receiving, or have received, corticoid treatment. Some of these patients may be unable to tolerate the stress of dental treatment, thus giving rise to the risk of acute adrenal cortical failure. The present study addresses the prevention of these unpleasant and hazardous situations, with a review of the different corticoid replacement therapies applicable in dental practice. PMID- 12730656 TI - Oral leiomyoma: a case report. AB - Oral leiomyoma is a benign smooth muscle tumour, that occurs most frequently in the uterine myometrium, gastrointestinal tract, and skin. Ocurrence in the oral cavity is considered rare, probably because of the paucity of smooth muscle tissue at this level. Smooth muscle tumours can occur at any age an usually is appear as a slow growing, firm mucosal nodule. Most lesions are asymptomatic, although occasional tumours can be painful. The most common sites are: lips, palate and tongue. The diagnosis of leiomyoma in the oral cavity is mainly determined by histological studies and special specific stains may be helpful to differentiate from other tumours, and also to confirm the smooth muscle origin is the diagnosis is in doubt. Surgical excision of the lesion appears to be the best treatment option. The purpose of this article is to present a case of a 62 year old woman, with a 6 month history of a leiomyoma in her right cheek and to explain its clinical features and treatment. PMID- 12730657 TI - Osseous choristoma of the oral soft tissue. Case report. AB - Oral osseous choristoma is a rare developmental alteration, their principal localization is nearly to base of the tongue (foramen caecum). The oral mucosa localization of osseous choristoma is extremely rare. At date only 10 cases of oral mucosa osseous choristoma had been reported. In the present paper we reported a new case of oral mucosa osseous choristoma in a Klippel-Feil syndrome patient. A review of available literature was made. We presented a 28 years old female patient who showed into the right oral mucosa, a hard, mobile, and asymptomatic mass, with minimum 4 years of evolution. The histological image showed a lesion constituted by lamellar bone, osteocytes and haematopoyetic tissue. The diagnosis of osseous choristoma was made. It is discusses their possible association with Klippel-Feil syndrome. The osseous choristoma of buccal mucosa is most frequently in fifth decade of the life although is reported between 12-to-64 years old, with a female predisposition. PMID- 12730658 TI - Mandibular metastasis of follicular thyroid carcinoma. Case report. AB - Thyroid carcinoma mandibular metastasis are not very frequent and the cases described in literature are few. Due to its bloodstream dissemination, most of them are a consequence of the follicular variant of thyroid carcinomas. A case is presented and a review of the clinicopathologic characteristics of the lesion is made, so the oral and maxilofacial surgeon can recognize it, make a correct differential diagnosis with other mandibular radioluciencies and in consequence, carry out an adequate treatment. PMID- 12730659 TI - [Clinical case. Pemphigus vulgaris]. PMID- 12730660 TI - [Treatment of oral Candidosis]. PMID- 12730661 TI - [Dental management of ischaemic heart disease]. PMID- 12730662 TI - [Dental management of patients with liver disease]. PMID- 12730663 TI - [Differential diagnosis of vesicle-erosive oral lesions]. PMID- 12730666 TI - Multiplexed genotyping with sequence-tagged molecular inversion probes. AB - We report on the development of molecular inversion probe (MIP) genotyping, an efficient technology for large-scale single nucleotide polymorphism (SNP) analysis. This technique uses MIPs to produce inverted sequences, which undergo a unimolecular rearrangement and are then amplified by PCR using common primers and analyzed using universal sequence tag DNA microarrays, resulting in highly specific genotyping. With this technology, multiplex analysis of more than 1,000 probes in a single tube can be done using standard laboratory equipment. Genotypes are generated with a high call rate (95%) and high accuracy (>99%) as determined by independent sequencing. PMID- 12730667 TI - High-throughput engineering of the mouse genome coupled with high-resolution expression analysis. AB - One of the most effective approaches for determining gene function involves engineering mice with mutations or deletions in endogenous genes of interest. Historically, this approach has been limited by the difficulty and time required to generate such mice. We describe the development of a high-throughput and largely automated process, termed VelociGene, that uses targeting vectors based on bacterial artificial chromosomes (BACs). VelociGene permits genetic alteration with nucleotide precision, is not limited by the size of desired deletions, does not depend on isogenicity or on positive-negative selection, and can precisely replace the gene of interest with a reporter that allows for high-resolution localization of target-gene expression. We describe custom genetic alterations for hundreds of genes, corresponding to about 0.5-1.0% of the entire genome. We also provide dozens of informative expression patterns involving cells in the nervous system, immune system, vasculature, skeleton, fat and other tissues. PMID- 12730668 TI - Modulation of p120E4F transcriptional activity by the Gam1 adenoviral early protein. AB - Gam1, an early adenoviral CELO protein, is required for viral replication. Consistent with its ability to inhibit histone deacetylation by HDAC1, Gam1 activates transcription. In this report, we identify the cellular transcription factor p120(E4F) as a Gam1 interaction partner. p120(E4F) is a low-abundance transcription factor that represses the adenovirus E4 promoter. Here we demonstrate that p120(E4F) interacts with HDAC1 in vivo and in vitro, and that E4F-mediated transcriptional repression is alleviated by the HDAC inhibitor trichostatin A or by overexpressing Gam1. A mutant E4 promoter unresponsive to E4F-mediated transcriptional repression is also not stimulated by Gam1. Moreover, our cofractionation experiments demonstrate that p120(E4F), HDAC1 and Gam1 may be concomitantly present in protein complexes. We conclude that Gam1 activates E4 dependent transcription possibly by inactivating HDAC1. PMID- 12730669 TI - Cell cycle inhibition mediated by the outer surface of the C/EBPalpha basic region is required but not sufficient for granulopoiesis. AB - CCAAT/enhancer binding protein alpha (C/EBPalpha) transactivates target genes dependent upon DNA binding via its basic region-leucine zipper domain and slows G1 progression by interaction with E2F, cdk2, or cdk4. E2F interacts with the non DNA-binding surface of the C/EBPalpha basic region and C/EBPalpha residues 1-70 are required for repressing E2F targets, while cdk2 and cdk4 bind residues 177 191. C/EBPalpha-ER induces the 32D cl3 myeloblast cell line to differentiate to granulocytes. C/EBPalpha-ER variants incapable of binding DNA slowed G1, but did not induce early or late granulopoiesis, indicating that cell cycle inhibition as mediated by C/EBPalpha is not sufficient for differentiation. C/EBPalpha-ER variants lacking residues 11-70 or residues 11-70 and 178-200 both slowed the G1 to S transition. C/EBPalpha(GZ)-ER, containing the GCN4 rather than the C/EBPalpha leucine zipper, also slowed G1. In contrast, C/EBPalpha(BRM2)-ER, carrying mutations in the outer surface of the basic region required for interaction with E2F, did not slow G1. C/EBPalpha(BRM2)-ER induced early markers of granulopoiesis much less efficiently than C/EBPalpha-ER and did not direct terminal maturation. Inhibition of G1 progression using mimosine increased induction of late markers by G-CSF. Thus, both DNA binding and cell cycle arrest, mediated by opposite surfaces of the C/EBPalpha basic region, are required for granulopoiesis. PMID- 12730670 TI - 2-Methoxyestradiol induces apoptosis in Ewing sarcoma cells through mitochondrial hydrogen peroxide production. AB - The Ewing sarcoma is the second most common bone tumor in children and young adults. Despite the advances in therapy, the 5-year survival rate for patients with metastatic disease is poor, indicating the need for alternative treatments. Here, we report that 2-methoxy-estradiol (2-Me), a natural estrogen metabolite, induced a caspase-dependent apoptosis of Ewing sarcoma-derived cells independently of their p53 status. 2-Me-induced apoptosis occurred through the mitochondrial death pathway as evidenced by reduction of the mitochondrial transmembrane potential, cytochrome c release and caspase-9 activation. Treatment of cells with 2-Me resulted in generation of intracellular H(2)O(2), which occurred earlier than caspase-9 activation. The H(2)O(2)-reducing agent Ebselen and the lipid peroxidation inhibitor vitamin E decreased both 2-Me-induced caspase-9 activation and cell death, thus providing evidence for a role of H(2)O(2) and lipid peroxides in the initiation of this process. Rotenone, an inhibitor of the mitochondrial respiratory chain, abolished both apoptosis and H(2)O(2) production, thereby identifying mitochondria as the source of H(2)O(2). Moreover, we observed that treatment of cells with 2-Me or H(2)O(2) induced activation of the c-Jun N-terminal kinase (JNK). Overexpression of a dominant negative mutant of JNK1 reduced 2-Me-induced apoptosis indicating that JNK participates in this process. Altogether, our results provide evidence that 2-Me triggers apoptosis of Ewing sarcoma cells through induction of a mitochondria redox-dependent mechanism and suggest that this compound or other agents that selectively increase the level of reactive oxygen species may prove useful to the development of novel strategies for treatment of Ewing tumors. PMID- 12730671 TI - Chemoprevention of mammary carcinogenesis in a transgenic mouse model by alpha difluoromethylornithine (DFMO) in the diet is associated with decreased cyclin D1 activity. AB - Mechanisms underlying the chemopreventive effect of difluoromethylornithine (DFMO) on the development of mammary cancer were investigated utilizing the whey acidic protein promoter-T antigen transgenic mouse model of breast cancer progression. Mice were exposed to four different doses of DFMO in the diet (3.5, 4.9, 7.0 and 10 g/kg diet). Tumor latency was increased in a dose-dependent manner. DFMO at the highest dose significantly delayed tumor onset (131 days as compared to 109 days in control unexposed mice, P=0.018). Analyses of preneoplastic mammary tissue collected 1 month after DFMO treatment demonstrated that DFMO (10 g/kg diet) significantly increased the ratio of apoptotic to proliferative indices (P=0.013) and significantly reduced the percentage of cells demonstrating nuclear localized cyclin D1 (P=0.013). Nuclear localizations of p27, p21 and Stat5a were not affected. Inhibitory effects of DFMO on cell growth and survival were lost as the cells progressed to cancer. In conclusion, the chemopreventive effects of DFMO on mammary cancer progression were mediated by changes in both apoptosis and cell proliferation in preneoplastic cells. Alterations in cyclin D1 activity in preneoplastic cells could represent an early biomarker of chemopreventive action and are consistent with a mechanistic role for cyclin D1 in progression of mammary cancer. PMID- 12730672 TI - Physical and functional interaction between HCV core protein and the different p73 isoforms. AB - Hepatitis C virus (HCV) core protein is a structural viral protein that packages the viral genomic RNA. In addition to this function, HCV core also modulates a number of cellular regulatory functions. In fact, HCV core protein has been found to modulate the expression of the cyclin-dependent inhibitor p21(WAF1/CIP1) and to promote both apoptosis and cell proliferation through its physical interaction with p53. Here, we studied the ability of HCV core to bind the p53-related p73 protein, its isoforms and its deletion mutants. We found that HCV core co immunoprecipitated with p73 in HepG2 and SAOS-2 cells. Deletion mutational analysis of p73 indicates that the domain involved in HCV core binding is located between amino-acid residues 321-353. We also demonstrate that p73/core interaction results in the nuclear translocation of HCV core protein either in the presence of the p73 alpha or p73 beta tumor-suppressor proteins. In addition, the interaction with HCV core protein prevents p73 alpha, but not p73 beta dependent cell growth arrest in a p53-dependent manner. Our findings demonstrate that HCV core protein may directly influence the various p73 functions, thus playing a role in HCV pathogenesis. PMID- 12730673 TI - Expression of the FAT10 gene is highly upregulated in hepatocellular carcinoma and other gastrointestinal and gynecological cancers. AB - The ubiquitin-like modifier (UBL) family has recently generated much interest in the scientific community, as it is implicated to play important regulatory roles via novel protein-protein modification. FAT10 (diubiquitin) belongs to this family of proteins, comprising two ubiquitin-like moieties fused in tandem, and has been implicated to be involved in the maintenance of spindle integrity during mitosis. As FAT10 may play a role in the regulation of genomic stability, we examined if there is an association between FAT10 expression and hepatocellular carcinoma (HCC) or other cancers. Northern blot analyses revealed upregulation of FAT10 expression in the tumors of 90% of HCC patients. In situ hybridization as well as immunohistochemistry utilizing anti-FAT10 antibodies localized highest FAT10 expression in the nucleus of HCC hepatocytes rather than the surrounding immune and non-HCC cells. FAT10 expression was also found to be highly upregulated in other cancers of the gastrointestinal tract and female reproductive system. In conclusion, we demonstrated upregulation of FAT10 expression in various gastrointestinal and gynecological cancers. Its overexpression is unrelated to the general increase in protein synthesis or a general immune/inflammatory response to cancer. Rather, FAT10 may modulate tumorigenesis through its reported interaction with the MAD2 spindle-assembly checkpoint protein. PMID- 12730674 TI - Integrity of c-Raf-1/MEK signal transduction cascade is essential for hepatitis B virus gene expression. AB - The genome of hepatitis B virus (HBV) encodes two transcriptional activators: the HBx protein and the PreS2-activator large surface protein (LHBs). Both proteins trigger activation of c-Raf-1/MEK kinase cascade. In case of HBx this can be mediated by a PKC-independent and Ras-dependent mechanism, in case of LHBs activation is PKC-dependent and does not require Ras. Selective destruction of either LHBs- or of HBx-specific activation does not result in significant decrease of viral production from transfected HepG2 cells. Simultaneous inhibition of LHBs- and HBx-dependent activation by blocking signaling steps common to both activators, using trans dominant negative c-Raf-1- or MEK-specific inhibitors, abolished HBV gene expression. In accordance with this no HBV propagation was observed after transfection of a mutated HBV genome defective for HBx- and PreS2-activator function. A detailed analysis revealed that the observed inhibition of HBV- propagation is because of a significant reduction of HBV specific RNA resulting in an inhibition of the de novo synthesis of viral compounds (viral proteins and nucleic acid) and not by blocking secretion or assembly of the virus. Based on these results we conclude that transcriptional activator function, mediated by the c-Raf-1/MEK signaling cascade, is essential for HBV gene expression. PMID- 12730675 TI - Phenoxodiol--an isoflavone analog--induces apoptosis in chemoresistant ovarian cancer cells. AB - Interference with the innate apoptotic activity is a hallmark of neoplastic transformation and tumor formation. In this study we characterize the cytotoxic effect of phenoxodiol, a synthetic anticancer drug analog of genestein, and demonstrate the mechanism of action by which phenoxodiol affects the components of the Fas apoptotic pathway on ovarian cancer cells. Primary ovarian cancer cells, isolated from ascitic fluids of ovarian cancer patients, resistant to conventional chemotherapy, undergo apoptosis following phenoxodiol treatment. This effect is dependent upon the activation of the caspase system, inhibiting XIAP, an inhibitor of apoptosis, and disrupting FLICE inhibitory protein (FLIP) expression through the Akt signal transduction pathway. We suggest that phenoxodiol is an efficient inducer of cell death in ovarian cancer cells and sensitizes the cancer cells to Fas-mediated apoptosis. We identified FLIP and XIAP signalling pathways as key factors regulating the survival of ovarian cancer cells. These findings demonstrate a novel nontoxic drug that controls FLIP/XIAP function and has the potential to eliminate tumor cells through Fas-mediated apoptosis. PMID- 12730676 TI - Intracellular mediators of erucylphosphocholine-induced apoptosis. AB - Induction of apoptosis contributes to the cytotoxic action of the intravenously applicable alkylphosphocholine erucylphosphocholine (ErPC). To define molecular requirements for ErPC-induced apoptosis, activation of caspases-8, -9 and -3 and cleavage of the caspase-3 substrates PARP and ICAD were tested in normal Jurkat T cells, Jurkat cells resistant to death receptor (CD95 or TNFalpha-related apoptosis inducing ligand (TRAIL)-induced apoptosis, Jurkat cells lacking caspase 8 or Fas-associated death domain (FADD) Jurkat cells expressing a dominant negative caspase-9 or overexpressing Bcl-2 as well as BJAB B-lymphoma cells expressing a dominant-negative FADD (FADD-DN). ErPC induced a time- and dose dependent apoptotic cell death in Jurkat and BJAB cells, which was characterized by breakdown of the phosphatidylserine asymmetry, depolarization of the mitochondrial membrane potential, release of cytochrome c, activation of caspases 9, -8 and -3, cleavage of PARP and ICAD, as well as chromatin condensation. ErPC induced apoptosis was independent from CD95-receptor signaling and FADD since CD95- and TRAIL-resistant, caspase-8- and FADD-negative Jurkat cells, as well as BJAB cells expressing FADD-DN were sensitive to ErPC-induced apoptosis. In contrast, inhibition of caspase-9 and overexpression of Bcl-2 significantly reduced ErPC-induced caspase activation and apoptosis. Thus, ErPC triggers apoptosis via a Bcl-2-dependent mitochondrial but death receptor-independent pathway. PMID- 12730677 TI - Expression of angiotensin type II receptor downregulates Cdk4 synthesis and inhibits cell-cycle progression. AB - Accumulating evidence suggests that angiotensin II type II (AT(2)) receptor subtype negatively regulates cell proliferation in pathophysiological conditions associated with tissue remodeling. However, the mechanisms through which AT(2) receptor achieves this effect remain poorly understood. In this study, we demonstrate that expression of AT(2) receptor inhibits the proliferation of rat fibroblasts in a ligand-independent manner. The antiproliferative action of AT(2) is dependent on the density of surface receptors. We show that AT(2) receptor expression negatively regulates G1 phase progression in both cycling cells and G0 arrested cells stimulated to re-enter the cell cycle, but has no detectable effect on apoptosis. The delay in cell-cycle progression of AT(2)-expressing cells is associated with downregulation of cyclin E expression, decreased assembly of cyclin E-Cdk2 complexes, and the resulting attenuation of Cdk2 activation. The induction of Cdk4 expression and activity is also markedly attenuated, which likely contributes to the inhibition of cyclin E expression. Ectopic expression of Cdk4 alleviates the proliferation defect of AT(2) expressing cells. These findings suggest that the growth-inhibitory effects of the AT(2) receptor are attributable in part to its spontaneous inhibitory action on the cell cycle machinery. PMID- 12730678 TI - Conformational change and mitochondrial translocation of Bax accompany proteasome inhibitor-induced apoptosis of chronic lymphocytic leukemic cells. AB - Chemotherapy resistance remains a major clinical problem in patients with B-cell chronic lymphocytic leukemia (B-CLL). Proteasome inhibitors are able to induce apoptosis in chemotherapy-resistant B-CLL cells in vitro. Exposure of B-CLL cells to the proteasome inhibitors, MG132 and lactacystin, resulted in inhibition of proteasomal activity within 30 min of treatment and was accompanied by an increase in the level of ubiquitinated proteins. Proteasome inhibitors did not alter the levels of expression of the proapoptotic Bcl-2 family proteins, Bax and Bid, prior to the onset of apoptosis. Instead, proteasome inhibitors induced a caspase-independent conformational change in Bax (as shown by a conformation specific Bax antibody) and its translocation to mitochondria, resulting in mitochondrial perturbation, as evidenced by loss of the mitochondrial membrane potential and cytochrome c release. Similar conformational change and subcellular localization of Bax were observed during apoptosis induced with fludarabine, chlorambucil and prednisolone. These data suggest that alteration of Bax conformation and its redistribution to mitochondria are common and early features of B-CLL apoptosis in response to proteasome inhibitors and other chemotherapeutic agents. PMID- 12730679 TI - N-terminal deletion augments the cell-death-inducing activity of BAX in adenoviral gene delivery to nonsmall cell lung cancers. AB - Therapeutic modalities that overcome the antiapoptotic function of Bcl-2 that is often overexpressed in cancer cells are expected to be a novel strategy for cancer treatment. We previously reported that the leukemic cell death induced by an N-terminally truncated Bax (deltaN Bax: corresponding to amino acid 112-192 of full-length Bax) was not blocked by Bcl-2 or Bcl-x(L) owing to the lack of the BH3 domain needed to interact with the antiapoptotic Bcl-2 family molecules. In this study, we used the Cre-loxP system that allowed us to propagate adenoviruses expressing deltaN Bax, and investigated the effects of the deltaN Bax gene transfer into A549 and NCI-H1299 nonsmall cell lung cancer cell lines. deltaN Bax showed more cell-death-inducing activity in both cells than did the full-length Bax in vitro. It was found that the deltaN Bax-induced cell death was not inhibited by the pan-caspase inhibitor z-VAD-fmk, suggesting that deltaN Bax induces cell death through a caspase-independent mechanism. Intratumoral injection of adenoviruses expressing deltaN Bax into A549 tumors in Balb/c nude mice showed a significantly stronger suppression of tumor growth (74%) than full length Bax (25%) compared to the control. Our results suggest that deltaN Bax may provide a better alternative than currently used cytotoxic genes in cancer gene therapy trials. PMID- 12730680 TI - N-Myc overexpression leads to decreased beta1 integrin expression and increased apoptosis in human neuroblastoma cells. AB - Neuroblastoma is a childhood tumor thought to arise through improper differentiation of neural crest cells. Increased N-Myc expression in neuroblastoma indicates highly malignant disease and poor patient prognosis. N myc enhances cell growth, insulin-like growth factor type I receptor (IGF-IR) expression, and tumorigenicity in combination with Bcl-2. Despite these effects, N-Myc overexpression in SHEP neuroblastoma cells (SHEP/N-Myc cells) increases serum-withdrawal and mannitol-induced apoptosis. Although we have previously shown a protective effect of IGF-I in SHEP cells, in SHEP/N-Myc cells IGF-I rescue from mannitol-induced apoptosis is prevented. N-Myc overexpression has little effect on IGF-IR signaling pathways, but results in increased Akt phosphorylation when Bcl-2 is coexpressed. A loss of integrin-mediated adhesion promotes apoptosis in many systems. SHEP/N-Myc cells have dramatically less beta1 integrin expression than control cells, consistent with previous reports. beta1 integrin expression is decreased in more tumorigenic neuroblastoma cells lines, including IMR32 and SH-SY5Y cells. Reintroduction of beta1 integrin into the N Myc-overexpressing cells prevents mannitol-mediated apoptosis. We speculate that N-Myc repression of beta1 integrin expression leads to a less differentiated phenotype, resulting in increased growth and tumorigenesis if properly supported or apoptosis if deprived of growth sustaining molecules. PMID- 12730681 TI - Effect of Bax deficiency on death receptor 5 and mitochondrial pathways during endoplasmic reticulum calcium pool depletion-induced apoptosis. AB - Thapsigargin (TG), by inducing perturbations in cellular Ca(2+) homeostasis, can induce apoptosis, but the molecular mechanisms remain to be fully elucidated. We have recently reported that TG-induced apoptosis appears to involve the DR5 dependent apoptotic pathway that cross talks with the mitochondrial pathway via TG-induced Bid cleavage. In this study, we have utilized Bax-proficient and deficient HCT116 human colon cancer cells to investigate the effect of Bax deficiency on TG-induced apoptosis and TG regulation of the DR5 and mitochondrial pathways. Our results indicate that Bax-deficient cells are less sensitive to undergo apoptosis following TG treatment. Our results further demonstrate that TG induced apoptosis is coupled with DR5 upregulation and caspases 8 and 3 activation, as well as Bid cleavage in both Bax-proficient and -deficient cells, although caspase 3 activation was reduced in Bax-deficient cells. TG also promoted the release of cytochrome c into cytosol and caspase 9 activation in Bax proficient cells but not in Bax-deficient cells. These findings suggest that although Bax is not absolutely required for death receptor (DR)-dependent signals, it appears to be a key molecule in TG-regulated mitochondrial events. Bax-deficient cells were relatively more resistant to Apo2L/TRAIL than the Bax proficient counterparts. However, the combination of Apo2L/TRAIL and TG was more effective in mediating apoptosis in both Bax-proficient and -deficient cells and that was coupled with activation of caspases 8 and 3. Although both agents in combination also induced cytochrome c release into cytosol and caspase 9 activation in Bax-proficient cells, these events were abrogated in Bax-deficient cells. Our results thus suggest that the combination of Apo2L/TRAIL and TG appears to bypass the Bax deficiency-induced defects in the mitochondrial (intrinsic) pathway by engaging the DR5-dependent apoptotic signals (extrinsic pathway). PMID- 12730682 TI - cDNA microarray analysis of genes associated with ERBB2 (HER2/neu) overexpression in human mammary luminal epithelial cells. AB - To investigate changes in gene expression associated with ERBB2, expression profiling of immortalized human mammary luminal epithelial cells and variants expressing a moderate and high level of ERBB2 has been carried out using cDNA microarrays corresponding to approximately 6000 unique genes/ESTs. A total of 61 significantly up- or downregulated (2.0-fold) genes were identified and further validated by RT-PCR analysis as well as microarray comparisons with a spontaneously ERBB2- overexpressing breast cancer cell line and ERBB2-positive primary breast tumors. The expression and clinical relevance of proteins predicted to be associated with ERBB2 overexpression in breast cancers were analysed together with their clinical relevance by antibody screening using a tissue array. Differentially regulated genes include those involved in cell matrix interactions including proline 4-hydroxylase (P4HA2), galectin 1 (LGALS1) and galectin 3 (LGALS3), fibronectin 1 (FN1) and p-cadherin (CDH3), and cell proliferation (CRIP1, IGFBP3) and transformation (S100P, S100A4). A number of genes associated with MYC signalling were also differentially expressed, including NDRG1, USF2 and the epithelial membrane proteins 1 and 3 (EMP1, EMP3). These data represent profiles of the transcriptional changes associated with ERBB2-related pathways in the breast, and identify novel and potentially useful targets for prognosis and therapy. PMID- 12730683 TI - Identification of H-Ras, RhoA, Rac1 and Cdc42 responsive genes. AB - The superfamily of small GTP-binding proteins has expanded dramatically in recent years. The Ras family has long been associated with signaling pathways contributing to normal and aberrant cell growth, while Rho-related protein function is to integrate extracellular signals with specific targets regulating cell morphology, cell aggregation, tissue polarity, cell motility and cytokinesis. Recent findings suggest that certain Rho proteins, including RhoA, Rac1 and Cdc42, can also play a role in signal transduction to the nucleus and cell growth control. However, the nature of the genes regulated by Ras and Rho GTPases, as well as their contribution to their numerous biological effects is still largely unknown. To approach these questions, we investigated the global gene expression pattern induced by activated forms of H-Ras, RhoA, Rac1 and Cdc42 using cDNA microarrays comprising 19 117 unique elements. Using this approach, we identified 1184 genes that were up- or downregulated by at least twofold. Hierarchical cluster analysis revealed the existence of patterns of gene regulation both unique and common to H-Ras V12, RhoA QL, Rac1 QL and Cdc42 QL activation. For example, H-Ras V12 upregulated osteopontin and Akt 1, and H-Ras and RhoA stimulated cyclin G1, cyclin-dependent kinase 8, cyclin A2 and HMGI-C, while Rac1 QL and Cdc42 QL upregulated extracellular matrix and cell adhesion proteins such as alpha-actinin 4, procollagen type I and V and neuropilin. Furthermore, H-Ras V12 downregulated by >eightfold 52 genes compared to only three genes by RhoA QL, Rac1 QL and Cdc42 QL. These results provide key information to begin unraveling the complexity of the molecular mechanisms underlying the transforming potential of Ras and Rho proteins, as well as the numerous morphological and cell cycle effects induced by these small GTPases.64 PMID- 12730684 TI - ATP-induced conformational changes of the nucleotide-binding domain of Na,K ATPase. AB - The Na,K-ATPase hydrolyzes ATP to drive the coupled extrusion and uptake of Na+ and K+ ions across the plasma membrane. Here, we report two high-resolution NMR structures of the 213-residue nucleotide-binding domain of rat alpha1 Na,K ATPase, determined in the absence and the presence of ATP. The nucleotide binds in the anti conformation and shows a relative paucity of interactions with the protein, reflecting the low-affinity ATP-binding state. Binding of ATP induces substantial conformational changes in the binding pocket and in residues located in the hinge region connecting the N- and P-domains. Structural comparison with the Ca-ATPase stabilized by the inhibitor thapsigargin, E2(TG), and the model of the H-ATPase in the E1 form suggests that the observed changes may trigger the series of events necessary for the release of the K+ ions and/or disengagement of the A-domain, leading to the eventual transfer of the gamma-phosphate group to the invariant Asp369. PMID- 12730685 TI - A novel mode of RBD-protein recognition in the Y14-Mago complex. AB - Y14 and Mago are conserved eukaryotic proteins that associate with spliced mRNAs in the nucleus and remain associated at exon junctions during and after nuclear export. In the cytoplasm, Y14 is involved in mRNA quality control via the nonsense-mediated mRNA decay (NMD) pathway and, together with Mago, is involved in localization of osk (oskar) mRNA. We have determined the crystal structure of the complex between Drosophila melanogaster Y14 and Mago at a resolution of 2.5 A. The structure reveals an atypical mode of protein-protein recognition mediated by an RNA-binding domain (RBD). Instead of binding RNA, the RBD of Y14 engages its RNP1 and RNP2 motifs to bind Mago. Using structure-guided mutagenesis, we show that Mago is also a component of the NMD pathway, and that its association with Y14 is essential for function. Heterodimerization creates a single structural platform that interacts with the NMD machinery via phylogenetically conserved residues. PMID- 12730686 TI - Structural insights into the catalytic mechanism of cyclophilin A. AB - Cyclophilins constitute a ubiquitous protein family whose functions include protein folding, transport and signaling. They possess both sequence-specific binding and proline cis-trans isomerase activities, as exemplified by the interaction between cyclophilin A (CypA) and the HIV-1 CA protein. Here, we report crystal structures of CypA in complex with HIV-1 CA protein variants that bind preferentially with the substrate proline residue in either the cis or the trans conformation. Cis- and trans-Pro substrates are accommodated within the enzyme active site by rearrangement of their N-terminal residues and with minimal distortions in the path of the main chain. CypA Arg55 guanidinium group probably facilitates catalysis by anchoring the substrate proline oxygen and stabilizing sp3 hybridization of the proline nitrogen in the transition state. PMID- 12730687 TI - Ultrafast enzymatic reaction dynamics in protochlorophyllide oxidoreductase. AB - The reaction catalyzed by the light-driven enzyme protochlorophyllide oxidoreductase (POR) has been initiated with a 50-fs laser pulse. We show that the catalytic mechanism, involving proton and hydride transfers, proceeds with time constants of 3 ps and 400 ps. It is known that catalysis by POR involves thermally excited protein dynamics; our results show that these molecular motions occur on an ultrafast timescale. PMID- 12730688 TI - Light-induced electron transfer in a cryptochrome blue-light photoreceptor. AB - Cryptochromes are flavoproteins implicated in multiple blue light-dependent signaling pathways regulating, for example, photomorphogenesis in plants or circadian clocks in animals. Using transient absorption spectroscopy, it is demonstrated that the primary light reactions in isolated Arabidopsis thaliana cryptochrome-1 involve intraprotein electron transfer from tryptophan and tyrosine residues to the excited flavin adenine dinucleotide cofactor. PMID- 12730689 TI - A nonpeptidyl mimic of superoxide dismutase, M40403, inhibits dose-limiting hypotension associated with interleukin-2 and increases its antitumor effects. AB - Interleukin-2 (IL-2) is used to treat metastatic renal cell carcinoma and malignant melanoma, but its use is limited by the severe hypotension it produces. We have shown here that M40403, a superoxide dismutase (SOD) mimetic, blocked IL 2-induced hypotension and allowed the dose of IL-2 to be increased in mice. The reversal of IL-2-mediated hypotension was associated with an increase in plasma catecholamines. In addition, M40403 increased lymphokine-activated killer (LAK) cell cytotoxicity in vitro and in vivo, through inhibition of macrophage superoxide production. Treatment of methylcholanthrene-induced (Meth A) ascites tumors with IL-2 and > or =3 mg per kg body weight M40403 induced 50% complete remissions lasting for more than 200 d, which was longer than those of untreated mice (15-d median survival) or mice treated with IL-2 alone (22-d median). Growth of subcutaneous implants of RENCA renal carcinoma was also inhibited by the combination of IL-2 and M40403. These results established that M40403 prevented IL-2 from causing dose-limiting hypotension, while enhancing its anticancer activity. PMID- 12730690 TI - Leukocytes mediate retinal vascular remodeling during development and vaso obliteration in disease. AB - Retinal ischemia can cause vision-threatening pathological neovascularization. The mechanisms of retinal ischemia are not fully understood, however. Here we have shown that leukocytes prune the retinal vasculature during normal development and obliterate it in disease. Beginning at postnatal day 5 (P5) in the normal rat, vascular pruning began centrally and extended peripherally, leaving behind a less dense, smaller-caliber vasculature. The pruning was correlated with retinal vascular expression of intercellular adhesion molecule-1 (ICAM-1) and coincided with an outward-moving wave of adherent leukocytes composed in part of cytotoxic T lymphocytes. The leukocytes adhered to the vasculature through CD18 and remodeled it through Fas ligand (FasL)-mediated endothelial cell apoptosis. In a model of oxygen-induced ischemic retinopathy, this process was exaggerated. Leukocytes used CD18 and FasL to obliterate the retinal vasculature, leaving behind large areas of ischemic retina. In vitro, T lymphocytes isolated from oxygen-exposed neonates induced a FasL-mediated apoptosis of hyperoxygenated endothelial cells. Targeting these pathways may prove useful in the treatment of retinal ischemia, a leading cause of vision loss and blindness. PMID- 12730691 TI - Subversion of the innate immune system by a retrovirus. AB - Retroviruses evolve rapidly to avoid the immune response of the infected host. We show here that the wild-type mouse mammary tumor virus MMTV(C3H) persisted indefinitely in C3H/HeN mice. However, it was rapidly lost in mice of the closely related C3H/HeJ strain and was replaced by a virus recombinant with an endogenous Mtv provirus. Maintenance of the wild-type virus was dependent on Toll-like receptor-4 (TLR4) signaling, which triggered production of the immunosuppressive cytokine interleukin-10. In the presence of mutant TLR4 in C3H/HeJ mice, wild type virus was eliminated by the cytotoxic immune response, promoting selection of the immune escape recombinant MMTV variants. Thus, subversion of the innate immune system is yet another survival strategy used by retroviruses. PMID- 12730692 TI - Dynamics of CD8+ T cell priming by dendritic cells in intact lymph nodes. AB - The cellular dynamics underlying activation of CD8+ T cells by dendritic cells (DCs) in the lymph node are not known. Here we have tracked the behavior of T cells and DCs by subjecting intact lymph nodes to real-time two-photon microscopy. We show that DCs scan at least 500 different T cells per hour in the absence of antigen. Antigen-bearing DCs are highly efficient in recruiting peptide-specific T cells and can engage more than ten T cells simultaneously. The duration of these interactions is of the order of hours, not minutes. The overall avidity of the interaction influences the probability that T cells will be stably captured by DCs, providing a possible basis for T cell competition. Taken together, our results identify the cellular behaviors that promote an efficient CD8+ T cell response in the lymph node. PMID- 12730693 TI - Deficiency of Mbd2 suppresses intestinal tumorigenesis. AB - Gene silencing through de novo methylation of CpG island promoters contributes to cancer. We find that Mbd2, which recruits co-repressor complexes to methylated DNA, is essential for efficient tumorigenesis in the mouse intestine. As Mbd2 deficient mice are viable and fertile, their resistance to intestinal cancer may be of therapeutic relevance. PMID- 12730695 TI - Alternative splicing in the human, mouse and rat genomes is associated with an increased frequency of exon creation and/or loss. AB - One of the most interesting opportunities in comparative genomics is to compare not only genome sequences but additional phenomena, such as alternative splicing, using orthologous genes in different genomes to find similarities and differences between organisms. Recently, genomics studies have suggested that 40-60% of human genes are alternatively spliced and have catalogued up to 30,000 alternative splice relationships in human genes. Here we report an analysis of 9,434 orthologous genes in human and mouse, which indicates that alternative splicing is associated with a large increase in frequency of recent exon creation and/or loss. Whereas most exons in the mouse and human genomes are strongly conserved in both genomes, exons that are only included in alternative splice forms (as opposed to the constitutive or major transcript form) are mostly not conserved and thus are the product of recent exon creation or loss events. A similar comparison of orthologous exons in rat and human validates this pattern. Although this says nothing about the complex question of adaptive benefit, it does indicate that alternative splicing in these genomes has been associated with increased evolutionary change. PMID- 12730694 TI - Transcription of antisense RNA leading to gene silencing and methylation as a novel cause of human genetic disease. AB - Nearly all human genetic disorders result from a limited repertoire of mutations in an associated gene or its regulatory elements. We recently described an individual with an inherited form of anemia (alpha-thalassemia) who has a deletion that results in a truncated, widely expressed gene (LUC7L) becoming juxtaposed to a structurally normal alpha-globin gene (HBA2). Although it retains all of its local and remote cis-regulatory elements, expression of HBA2 is silenced and its CpG island becomes completely methylated early during development. Here we show that in the affected individual, in a transgenic model and in differentiating embryonic stem cells, transcription of antisense RNA mediates silencing and methylation of the associated CpG island. These findings identify a new mechanism underlying human genetic disease. PMID- 12730696 TI - Different noses for different people. AB - Of more than 1,000 human olfactory receptor genes, more than half seem to be pseudogenes. We investigated whether the most recent of these disruptions might still segregate with the intact form by genotyping 51 candidate genes in 189 ethnically diverse humans. The results show an unprecedented prevalence of segregating pseudogenes, identifying one of the most pronounced cases of functional population diversity in the human genome. PMID- 12730697 TI - Mutations in PCSK9 cause autosomal dominant hypercholesterolemia. AB - Autosomal dominant hypercholesterolemia (ADH; OMIM144400), a risk factor for coronary heart disease, is characterized by an increase in low-density lipoprotein cholesterol levels that is associated with mutations in the genes LDLR (encoding low-density lipoprotein receptor) or APOB (encoding apolipoprotein B). We mapped a third locus associated with ADH, HCHOLA3 at 1p32, and now report two mutations in the gene PCSK9 (encoding proprotein convertase subtilisin/kexin type 9) that cause ADH. PCSK9 encodes NARC-1 (neural apoptosis regulated convertase), a newly identified human subtilase that is highly expressed in the liver and contributes to cholesterol homeostasis. PMID- 12730698 TI - A new phospholipid phosphatase, PRG-1, is involved in axon growth and regenerative sprouting. AB - Outgrowth of axons in the central nervous system is governed by specific molecular cues. Molecules detected so far act as ligands that bind to specific receptors. Here, we report a new membrane-associated lipid phosphate phosphatase that we have named plasticity-related gene 1 (PRG-1), which facilitates axonal outgrowth during development and regenerative sprouting. PRG-1 is specifically expressed in neurons and is located in the membranes of outgrowing axons. There, it acts as an ecto-enzyme and attenuates phospholipid-induced axon collapse in neurons and facilitates outgrowth in the hippocampus. Thus, we propose a novel mechanism by which axons are able to control phospholipid-mediated signaling and overcome the growth-inhibiting, phospholipid-rich environment of the extracellular space. PMID- 12730699 TI - Parietal activity and the perceived direction of ambiguous apparent motion. AB - We recorded from parietal neurons in monkeys (Macacca mulatta) trained to report the direction of an apparent motion stimulus consisting of regularly spaced columns of dots surrounded by an aperture. Displacing the dots by half their inter-column spacing produced vivid apparent motion that could be perceived in either the preferred or anti-preferred direction for each neuron. Many neurons in the lateral intraparietal area (LIP) responded more strongly on trials in which the animals reported perceiving the neurons' preferred direction, independent of the hand movement used to report their percept. This selectivity was less common in the medial superior temporal area (MST) and virtually absent in the middle temporal area (MT). Variations in activity of LIP and MST neurons just before motion onset were also predictive of the animals' subsequent perceived direction. These data suggest a hierarchy of representation in parietal cortex, whereby neuronal responses become more aligned with subjective perception in higher parietal areas. PMID- 12730700 TI - Synchrony-dependent propagation of firing rate in iteratively constructed networks in vitro. AB - The precise role of synchronous neuronal firing in signal encoding remains unclear. To examine what kinds of signals can be carried by synchrony, I reproduced a multilayer feedforward network of neurons in an in vitro slice preparation of rat cortex using an iterative procedure. When constant and time varying frequency signals were delivered to the network, the firing of neurons in successive layers became progressively more synchronous. Notably, synchrony in the in vitro network developed even with uncorrelated input, persisted under a wide range of physiological conditions and was crucial for the stable propagation of rate signals. The firing rate was represented by a classical rate code in the initial layers, but switched to a synchrony-based code in the deeper layers. PMID- 12730701 TI - Lonidamine: efficacy and safety in clinical trials for the treatment of solid tumors. AB - Lonidamine, a derivate of indazole-3-carboxylic acid, is an antineoplastic drug with a typical mechanism of action. Lonidamine has no function on cellular nucleic acids or protein synthesis, whereas it exerts a powerful inhibitory effect on oxygen consumption, aerobic glycolysis and lactate transport and accumulation of neoplastic cells. Nevertheless, its proven ability to modify the permeability of membranes is consistent with the possible increase of drug uptake, reverse of drug resistance and triggering of apoptotic pathway. Lonidamine has been experimentally shown to potentiate the cytotoxic effects of anthracyclines in human breast cancer cell lines and cisplatin activity in both platinum-sensitive and platinum-resistant human ovarian carcinoma cell lines. Since the specific mechanism of action and side effects are not overlapping with those of standard antineoplastic agents, combination of lonidamine with standard chemotherapy has been widely investigated for the treatment of solid tumors. Additionally, the enhancement of radiotherapy activity by lonidamine has been considered for palliative therapy of lesions from metastatic cancers. The encouraging results of phase II-III trials for the treatment of advanced breast, ovarian and lung cancer must be confirmed by larger studies. Specifically designed studies to address the role of lonidamine in the adjuvant setting are warranted. Lonidamine, a dechlorinate derivative of indazole-3-carboxylic acid, has proved to exert a powerful antiproliferative effect and to impair the energy metabolism of neoplastic cells. Herein we review the current experience on combining lonidamine and chemotherapy and/or radiation therapy in the treatment of solid tumors. Several studies have been published on this topic. The total number of trials reported in literature and length of follow-up are still insufficient to draw a firm conclusion. However, the available data demonstrate a significant role of lonidamine in modulating anthracycline and platinum compound activity. PMID- 12730702 TI - Association between homocyst(e)ine levels and risk of vascular events. AB - Homocyst(e)ine is a novel risk factor in vascular disease. First observations of vascular lesions in children with high blood homocyst(e)ine levels due to severe inborn enzyme deficiencies led to the hypothesis that elevated blood homocyst(e)ine levels might be a risk factor for vascular disease. A substantial body of evidence on the role of the homocyst(e)ine in the development of coronary and carotid artery disease, myocardial infarction, stroke, deep vein thrombosis and other disorders has been accumulated over the last 30 years. Cross-sectional and case-control studies provide initial and the strongest support for the hypothesis, followed by results from the prospective cohorts. Infrequent cases of homozygous mutations of the key enzymes in the homocyst(e)ine metabolism chain are able to produce extreme homocyst(e)inemia and early vascular lesions. More frequently, heterozygous enzyme mutations and deficiencies of folate and vitamins B6 and B12 cause mild to moderate homocyst(e)inemia, which is still strongly associated with the increased risk of vascular events. Elevated homocyst(e)ine levels may be effectively managed with adequate folate, B12 and B6 intake in doses comparable to or above FDA recommendations. Whether correction of elevated homocyst(e)ine levels with vitamins is helpful in prevention and treatment of vascular events remains unknown and is under investigation in ongoing clinical trials (VISP, VITATOPS). No consensus on homocyst(e)ine management is available at the present time. PMID- 12730703 TI - The etiology of erectile dysfunction and mechanisms by which drugs improve erection. AB - Following the National Institutes of Health (NIH) consensus conference in 1988, erectile dysfunction is defined as the consistent inability to maintain a penile erection sufficient for adequate sexual relations (1). The advances in basic and clinical research during the last two decades have led to the development of several new treatment options for erectile dysfunction, including new pharmacological agents for intracavernosal, intraurethral and oral use. The recent advent of medical therapy and the poor results of long-term follow-up in reconstructive vascular surgery, have significantly modified the medical management of this disorder. Discussion of erectile dysfunction has increased, information about erectile dysfunction is increasingly available, training in erectile dysfunction was improved and last, but not least, the number of patients seeking help for erectile dysfunction is growing, because satisfactory sexual function is an important part of a couple's healthy relationship and ongoing quality of life. PMID- 12730704 TI - Selective COX-2 inhibitors as chemopreventive and therapeutic agents. AB - Selective cyclooxygenase-2 (COX-2) inhibitors have received increasing attention for their role in the prevention and treatment of cancer. Considerable preclinical data support this use. Furthermore, clinical studies have shown that this enzyme is upregulated in a variety of premalignant and malignant states and that its inhibition can decrease colon polyp formation in patients with familial adenomatous polyposis. A number of studies are now investigating the use of COX-2 inhibitors to prevent or treat a number of different cancers. These ongoing trials will determine whether these agents are useful in the treatment of cancer. PMID- 12730705 TI - Current status and perspectives on the development of neuroprotectants for ischemic cerebrovascular disease. AB - The concept of neuroprotection is based on the understanding that delayed neuronal damage occurs after ischemia. Each step along the ischemic cascade provides an opportunity for therapeutic intervention. Based on the excellent results obtained in experimental models of ischemia, many clinical trials have been conducted with different neuroprotective drugs. The results obtained in most studies were negative and in some cases the studies were terminated due to safety problems. However, the mechanisms that underlie the development of ischemic damage are still being discovered, creating new therapeutic possibilities for neuroprotection that might be clinically applicable in the future. This article reviews the mechanisms of ischemic neuronal injury, the mechanism of action of neuroprotective agents, current neuroprotective clinical trials, and probable reasons for the failure of clinical neuroprotection. (c) 2003 Prous Science. All rights reserved. PMID- 12730706 TI - The immune system and microorganisms: a love-hate relationship revisited. PMID- 12730707 TI - Enteric pathogens and soil: a short review. AB - It is known that soil is a recipient of solid wastes able to contain enteric pathogens in high concentrations. Although the role of soil as a reservoir of certain bacterial pathogens is not in question, recent findings show that soil may have a larger role in the transmission of enteric diseases than previously thought. Many of the diseases caused by agents from soil have been well characterized, although enteric diseases and their link to soil have not been so well studied. Gastrointestinal infections are the most common diseases caused by enteric bacteria. Some examples are salmonellosis ( Salmonella sp.), cholera ( Vibrio cholerae), dysentery ( Shigella sp.) and other infections caused by Campylobacter jejuni, Yersinia sp. and Escherichia coli O157:H7 and many other strains. Viruses are the most hazardous and have some of the lowest infectious doses of any of the enteric pathogens. Hepatitis A, hepatitis E, enteric adenoviruses, poliovirus types 1 and 2, multiple strains of echoviruses and coxsackievirus are enteric viruses associated with human wastewater. Among the most commonly detected protozoa in sewage are Entamoeba histolytica, Giardia intestinalis and Cryptosporidium parvum. This article reviews the existing literature of more than two decades on waste disposal practices that favor the entry of enteric pathogens to soil and the possible consequent role of the soil as a vector and reservoir of enteric pathogens. PMID- 12730708 TI - Genetic basis of microbial carotenogenesis. AB - The synthesis of carotenoids begins with the formation of a phytoene from geranylgeranyl pyrophosphate, a well conserved step in all carotenogenic organisms and catalyzed by a phytoene synthase, an enzyme encoded by the crtB ( spy) genes. The next step is the dehydrogenation of the phytoene, which is carried out by phytoene dehydrogenase. In organisms with oxygenic photosynthesis, this enzyme, which accomplishes two dehydrogenations, is encoded by the crtP genes. In organisms that lack oxygenic photosynthesis, dehydrogenation is carried out by an enzyme completely unrelated to the former one, which carries out four dehydrogenations and is encoded by the crtI genes. In organisms with oxygenic photosynthesis, dehydrogenation of the phytoene is accomplished by a zeta carotene dehydrogenase encoded by the crtQ ( zds) genes. In many carotenogenic organisms, the process is completed with the cyclization of lycopene. In organisms exhibiting oxygenic photosynthesis, this step is performed by a lycopene cyclase encoded by the crtL genes. In contrast, anoxygenic photosynthetic and non-photosynthetic organisms use a different lycopene cyclase, encoded by the crtY ( lyc) genes. A third and unrelated type of lycopene beta cyclase has been described in certain bacteria and archaea. Fungi differ from the rest of non-photosynthetic organisms in that they have a bifunctional enzyme that displays both phytoene synthase and lycopene cyclase activity. Carotenoids can be modified by oxygen-containing functional groups, thus originating xanthophylls. Only two enzymes are necessary for the conversion of beta-carotene into astaxanthin, using several ketocarotenoids as intermediates, in both prokaryotes and eukaryotes. These enzymes are a beta-carotene hydroxylase ( crtZ genes) and a beta-carotene ketolase, encoded by the crtW (bacteria) or bkt (algae) genes. PMID- 12730709 TI - Fungus propagules in plastids: the mycosome hypothesis. AB - A stress-induced "mycosome" phase of Aureobasidium pullulans consisting of minute reproductive propagules that may revert directly to walled yeast cells is described. Mycosomes detected by light- and electron-microscopy reproduce within senescent plant plastids, and display three developmental pathways: wall-less cells (protoplasts), yeast cells, or membrane-bounded spherules that harbor plastids. Widespread in plant and algal cells, mycosomes are produced by both ascomycete and basidiomycete fungi. PMID- 12730710 TI - A methodological approach to investigate steady state fucoxanthin chlorophyll a/c binding protein mRNA levels in Wadden Sea sediments. AB - A method was established to investigate the steady state levels of mRNAs from genes encoding fucoxanthin chlorophyll a/c binding proteins (Fcp) of diatoms in situ. During the study, which was performed with Wadden Sea sediments from the German North Sea shore near Dangast, oxygenic photosynthesis was carried out mainly by pennate diatoms. Field samples were taken after tidal exposure from dawn up to late afternoon at 2-hourly intervals, and frozen in liquid nitrogen. In the laboratory, total RNA was isolated by isopycnic ultracentrifugation in caesium chloride gradients. Yields of approximately 10-300 micro g RNA per gram wet sediment were obtained. Defined amounts of total RNA were blotted onto nylon membranes and hybridised with probes against the fcp2 and 18S rDNA genes of Cyclotella cryptica. To estimate the steady state amount of fcp mRNAs, fcp signal intensities were normalized to the signal intensities obtained from hybridisation to an 18S rDNA gene probe. In the two time-course studies performed to demonstrate the applicability of the method, the steady state levels of fcp mRNA increased up to 12-fold with the onset of light, reaching a maximum 6-8 h after sunrise before they decreased again. Possible reasons for this time-course are discussed. PMID- 12730711 TI - Combination ecotoxicity and testing of common chemical discharges to sewer using the Vibrio fischeri luminescence bioassay. AB - In order to investigate possible synergistic or antagonistic (more or less than additive) toxicity effects, mixtures of chemicals were tested in water using a microbial bioassay. Ten toxicants (3,4-dichloroaniline, 3,5-dichlorophenol, cadmium, chromium, copper, Lindane, linear alkylbenzene sulphonate, pentachlorophenol, toluene, zinc) were chosen on the basis of their common occurrence in industrial effluents within local waste water treatment plants. These toxicants also cover a wide range of modes of toxic action, namely, polar and non-polar narcosis, membrane disruption, respiratory disruption, uncouplers of oxidative phosphorylation, biochemical disruption and enzyme inhibition. Efficient screening for possible combination toxicity between toxicants involved testing the chemicals both singly and in triplet combinations. The triplets were based on four replicates of a balanced incomplete block design (BIB). A standardised Vibrio fischeri rapid toxicity bioluminescence assay was used. The combinations tested showed that only one mixture was found to be significantly more toxic than expected from the pure single-toxicant results. Two triplets were significantly less toxic. Further tests on the more toxic triplet showed that the effect was due to only one of the 45 pairs originally screened. It is concluded that synergistic effects in combinations of toxicants are rather rare in bioluminescence systems utilising common effluents discharged to sewer. PMID- 12730712 TI - Intracellular transport of a heterologous membrane protein, the human transferrin receptor, in Saccharomyces cerevisiae. AB - We have analyzed the intracellular behavior of the human transferrin receptor (TfR) in Saccharomyces cerevisiae. The major part of the heterologously expressed TfR, which has previously been used as a model for heterologous expression of membrane proteins in yeast, is localized in the endoplasmic reticulum (ER) membranes; a minor fraction is present in the plasma membrane (PM). The stability of the TfR depends on vacuolar proteases, implying that it is degraded in the vacuolar compartment. Degradation is further dependent on favorable transport conditions to this compartment. The main bottleneck of transport seems to be the transition from the ER to the PM. The chaperone Cne1p, which is involved in quality control in the ER, plays a role in regulating the amount of heterologous TfR, as deletion of CNE1 leads to significant accumulation of the protein. This is the first demonstration of the involvement of CNE1 in regulating the level of heterologous membrane proteins. PMID- 12730713 TI - Conjugative plasmid mediated inducible nickel resistance in Hafnia alvei 5-5. AB - Hafnia alvei 5-5, isolated from a soil-litter mixture underneath the canopy of the nickel-hyperaccumulating tree Sebertia acuminata (Sapotaceae) in New Caledonia, was found to be resistant to 30 mM Ni(2+) or 2 mM Co(2+). The 70-kb plasmid, pEJH 501, was transferred by conjugation to Escherichia coli, Serratia marcescens, and Klebsiella oxytoca. Transconjugant strains expressed inducible nickel resistance to between 5 and 17 mM Ni(2+), and cobalt resistance to 2 mM Co(2+). A 4.8-kb Sal- EcoRI fragment containing the nickel resistance determinant was subcloned, and the hybrid plasmid was found to confer a moderate level of resistance to nickel (7 mM Ni(2+)) even to E. coli. The expression of nickel resistance was inducible by exposure to nickel chloride at a concentration as low as 0.5 mM Ni(2+). By random Tn phoA'-1 insertion mutagenesis, the fragment was shown to have structural genes as well as regulatory regions for nickel resistance. Southern hybridization studies showed that the nickel-resistance determinant from pEJH501 of H. alvei 5-5 was homologous to that of pTOM9 from Alcaligenes xylosoxydans 31A. PMID- 12730714 TI - News from Cairo: 18th International Conference on Arginine and Pyrimidines (ICAP 2002; 13-16 October 2002). PMID- 12730717 TI - Isolation and identification of a novel cDNA that encodes human yrdC protein. AB - In the course of detecting the interaction protein of RBBP10 by yeast two hybridization, we isolated a novel cDNA that encodes a putative human protein with yrdC domain. It is named human yrdC protein. Because the cDNA contains an open reading fragment (ORF) without a 5' in- frame stop codon, 5' RACE and 3' RACE were proceeded to produce the full-length cDNA. An 1825 bp cDNA was isolated from human placenta, which encodes a putative protein of 279 amino acids. The protein contains a sua5-yciO-yrdC domain. Blast analysis against the human genome database of Genbank revealed that the gene contains five exons, and assigned the gene to human chromosome 1p34.2. A transcript about 2.5 kb is ubiquitously expressed in human tissues. The gene is highly conserved during evolution. PMID- 12730718 TI - High-density SNP map of human ITR, a gene associated with vascular remodeling. AB - We constructed a high-density map of single-nucleotide polymorphisms (SNPs) present within a 31-kb region of human chromosome 13q31 that contains the human counterpart of the rabbit ITR gene, which encodes a rhodopsin-like G protein coupled receptor associated with vascular remodeling. The elements of human ITR cDNA were distributed in 27,452 bp of genomic DNA; the nine exons ranged in size from 50 bp to 2271 bp, with an average size of 392 bp. We isolated a total of 22 SNPs from the ITR locus by systematically screening genomic DNA from 48 healthy Japanese individuals; three SNPs were present in the 5' flanking region, two in coding elements, 12 in introns, and five in the 3' untranslated region. By comparing our data with SNPs deposited in the dbSNP database in the National Center for Biotechnology Information, 19 of the 22 SNPs (86%) were considered to be novel. The map presented here should help in evaluating the role of human ITR in cardiovascular diseases, in other diseases mapped to this segment on chromosome 13q31, and in a variety of pharmacogenetic effects. PMID- 12730720 TI - Expression profiles of two types of human knee-joint cartilage. AB - We have performed a comprehensive analysis of gene-expression profiles in human articular cartilage (hyaline cartilage) and meniscus (fibrocartilage) by means of a cDNA microarray consisting of 23,040 human genes. Comparing the profiles of the two types of cartilage with those of 29 other normal human tissues identified 24 genes that were specifically expressed in both cartilaginous tissues; these genes might be involved in maintaining phenotypes common to cartilage. We also compared the cartilage profiles with gene expression in human mesenchymal stem cells (hMSC), and detected 22 genes that were differentially expressed in cells representing the two cartilaginous lineages, 11 specific to each type, which could serve as markers for predicting the direction of chondrocyte differentiation. Our data should also provide useful information about regeneration of cartilage, especially in support of efforts to identify cartilage specific molecules as potential agents for therapeutic approaches to joint repair. PMID- 12730719 TI - A pair of sibs with tibial hemimelia born to phenotypically normal parents. AB - Tibial hemimelia is a rare congenital anomaly characterized by deficiency of the tibia with relatively intact fibula. Tibial hemimelia is identified as a solitary disorder, or a part of more complex malformation syndromes. Although the majority of cases with tibial hemimelia are sporadic, affected families with possible autosomal dominant or autosomal recessive inheritance have been reported. Here we report a pair of sibs, 6- and 2-year-old Japanese boys, with tibial hemimelia born to unrelated, phenotypically normal parents. The type of tibial hemimelia and associated malformations of hands and feet was quite different between the brothers. The elder brother was compatible with the Gollop-Wolfgang complex, and the younger brother with tibial agenesis-ectrodactyly syndrome. Screening of mutation by direct sequencing of candidate genes including Sonic hedgehog, HOXD 11, and HOXD-12 was unable to identify a disease-causing mutation. PMID- 12730721 TI - Lewis enzyme (alpha1-3/4 fucosyltransferase) polymorphisms do not explain the Lewis phenotype in the gastric mucosa of a Portuguese population. AB - The human alpha-1,3/4 fucosyltransferase III (FucT III) catalyses the synthesis of Lewis antigens including Le(b) antigen which is a ligand for Helicobacter pylori adhesion. Several polymorphisms have been described in the FUT3 gene affecting both the transmembrane and catalytic domains, some of which affect the enzyme activity. The aim of the present work was to study the Lewis gene polymorphisms in a Caucasian Portuguese population, with a high rate of H. pylori infection, and to evaluate the implications of mutant enzymes in Le(b) expression in the gastric mucosa. We studied 460 asymptomatic or dyspeptic individuals from northern Portugal. Screening for Lewis gene polymorphisms was performed by SSCP and direct sequencing. Lewis phenotype in gastric mucosa was determined by immunohistochemistry. In 47 individuals with a Lewis negative blood group, we found FUT3 gene polymorphisms that were previously described in other populations: 59T>G, 202T>C, 314C>T, 508G>A and 1067T>A. Among the 47 Lewis negative individuals in blood, only nine were also negative in gastric mucosa, suggesting the existence of another alpha 1-4 fucosyltransferase that is responsible for Le(a) and Le(b) synthesis in gastric mucosa. PMID- 12730722 TI - Analysis of the MTHFR 1298A-->C and 677C-->T polymorphisms as risk factors for neural tube defects. AB - The thermolabile variant (677TT) of methylenetetrahydrofolate reductase (MTHFR) is a known risk factor for neural tube defects (NTDs). The relationship between a second MTHFR polymorphism (1298A-->C) and NTD risk has been inconsistent between studies. We genotyped 276 complete NTD triads (mother, father and child affected with an NTD) and 256 controls for MTHFR 1298A-->C. Our findings do not support a role for the 1298A-->C polymorphism in NTDs (OR 0.85 (95% CI 0.49-1.47), p= 0.55), nor do we observe a combined effect with the 677C-->T polymorphism. PMID- 12730723 TI - Predominant maternal expression of the mouse Atp10c in hippocampus and olfactory bulb. AB - The human chromosome 15q11-q13 region is one of the most intriguing imprinted domains, and the abnormalities inherited are associated with neurological disorders including Prader-Willi syndrome (PWS), Angelman syndrome (AS) and autism. Recently we have identified a novel maternally expressed gene, ATP10C, that encodes a putative aminophospholipid translocase within this critical region, 200 kb distal to UBE3A in an imprinted domain on human chromosome 15. ATP10C, with UBE3A, displayed tissue-specific imprinting with predominant expression of the maternal allele in the brain. In this study, we demonstrated that the mouse homologue, Atp10c/pfatp, showed tissue-specific maternal expression in the hippocampus and olfactory bulb, which overlapped the region of imprinted Ube3a expression. These data suggest that the imprinted transcript of Atp10c in the specific region of CNS may be associated with neurological disorders including AS and autism. PMID- 12730725 TI - Identification of pathogenic mutations in the human rapsyn gene. AB - Rapsyn, a complex postsynaptic protein of the striated muscle, assembles acetylcholine receptors (AChR) at high density at the motor endplate (EP). Neuromuscular junctions of mice lacking rapsyn show no clusters of AChRs or other structural postsynaptic proteins such as beta-dystroglycan and utrophin. Humans with mutations in the rapsyn gene ( RAPSN) are affected with a postsynaptic form of congenital myasthenic syndrome (CMS) characterized by impairment of the morphologic development of the postsynaptic region. We have identified four patients from four different families with RAPSNmutations and CMS, confirmed in two cases by microelectrode and electron microscopy studies. The N88K mutation was present in all patients. One patient who was homozygous for N88K was only mildly affected, while the other three patients who were heterozygous for N88K and a second mutation (either L14P, 46insC, or Y269X) were severely affected. Mutations 46insC and Y269X predicts truncation of the protein. L14P predicts a conformational change at the N-terminus that may disrupt membrane association. N88K occurs within the putative leucine zipper motif potentially important for AChR clustering. These findings may explain the severe clinical involvement of compound heterozygous patients. PMID- 12730724 TI - Familial hypercholesterolemia in Morocco: first report of mutations in the LDL receptor gene. AB - Familial hypercholesterolemia (FH) is a genetic disorder mainly caused by defects in the low-density lipoprotein receptor (LDLR) gene, although it can also be due to alterations in the gene encoding apolipoprotein B (familial defective apoB or FDB) or in other unidentified genes. In Morocco, the molecular basis of FH is unknown. To obtain information on this issue, 27 patients with FH from eight unrelated families were analyzed by screening the LDLR (PCR-SSCP and Southern blot) and apoB genes (PCR and restriction enzyme digestion analysis). None of the patients carried either the R3500Q or the R3531C mutation in the apoB gene. By contrast, seven mutations in the LDLR gene were identified, including five missense mutations on exons 4, 6, 8, and 14 (C113R, G266C, A370T, P664L, C690S) and two large deletions (FH Morocco-1 and FH Morocco-2). The two major rearrangements and the missense mutation G266C are novel mutations and could well be causative of FH in the Moroccan population. This study has yielded preliminary information on the mutation spectrum of the LDLR gene among patients with FH in Morocco. PMID- 12730726 TI - Calcr, a brain-specific imprinted mouse calcitonin receptor gene in the imprinted cluster of the proximal region of chromosome 6. AB - Expressed sequence tags (ESTs) in the human chromosome 7q21-q31 region were recently used to screen for allelic expression bias in monochromosomal hybrids retaining a paternal or maternal human chromosome 7. Six candidate imprinted genes were identified. In this study, we investigated parent-of-origin-specific expression profiles of their mouse homologues in the proximal region of chromosome 6. An imprinting analysis, using F1 mice from reciprocal crosses between the B6 and JF strains, demonstrated that the mouse calcitonin receptor gene ( Calcr) was expressed preferentially from the maternal allele in brain, whereas no allelic bias was detected in other tissues. Our results indicate that Calcr is imprinted in a tissue-specific manner, with a predominant expression from the maternal allele in the brain. PMID- 12730727 TI - Setting up a palliative care clinic within a radiotherapy department: a model for developing countries. AB - Nearly 50% of all newly diagnosed cancer patients in India (and other developing countries) are terminally ill with advanced disease. These patients are usually neglected or often receive futile anticancer treatment(s), whereas what they really need is maximum medical management in the form of palliative care and psychosocial support. Since advanced and incurable cancer cases are mostly referred for radiotherapy (RT), a palliative care (PC) clinic was started in the Department of Radiotherapy, PGIMER, Chandigarh. The PC clinic staff consisted of one specialist doctor, a nurse and volunteers. Previous disease and treatment records maintained by the RT colleagues were noted. Proforma-based assessments were done in the PC clinic and focused on patients' Karnofsky Performance Status, physical symptoms, drugs prescribed, and the doctor's or patient's/relative's response to/satisfaction with the treatment in each case. Prospective data on 100 patients (March to August 2001) revealed that various distressing physical symptoms (cachexia, dyspnoea, constipation) had not been routinely assessed earlier. Despite previous treatment, adequate pain management as per the WHO ladder was needed in 67 of 88 (76%) patients when they were seen by the PC team. On the regular follow-up visits to the PC clinic, 42% and 50% of the patients/relatives reported a response to and satisfaction with the treatment at their second and third visits. We believe two conclusions are justified. (1) Attention to palliative care needs could result in good treatment outcome and high level of patients' and doctors' satisfaction. (2) Since a specialist PC set up is lacking in most medical institutions in India, the RT department is the best suited to delivery of palliative care for patients with advanced cancer. PMID- 12730728 TI - Factors predicting fatigue in breast cancer patients. AB - A prospective study was designed to investigate the factors predicting fatigue in breast cancer patients using the Cancer Fatigue Scale (CFS) in addition to the Hospital Anxiety and Depression Scale (HADS) and a questionnaire containing items on demographic and clinical data, and measures of patients' physical symptoms. The CFS measures total fatigue score ranging from 0 (lowest level) to 60 (highest level) and contains three subscales namely: physical, affective and cognitive fatigue. The questionnaires were administered to a consecutive sample of breast cancer patients attending the Iranian Center for Breast Cancer either for their treatment or follow-up examination. In all, 112 patients were studied. The mean age of the respondents was 45.7 years (SD11.0). Most had stage II breast cancer (67%) and had completed their initial treatment (45%). The mean total fatigue score of the patients was 18.7 (SD 13.5) and overall 49% reported experiencing fatigue to some degree (from quite a bit to very much). Severe anxiety and depression was reported by 16% and 32%, respectively. The regression analysis revealed that fatigue was predicted by depression ( P=0.003), pain ( P=0.005), current tamoxifen use ( P=0.001), undergoing mastectomy ( P=0.03) and anxiety ( P=0.04). The other variables studied did not emerge as significant predictors of fatigue in the regression analysis. The study findings suggest that in comparison to treatment factors, physical and psychological symptoms have a more important role in cancer-related fatigue and that fatigue should be recognized and managed even before commencing breast cancer treatment. PMID- 12730730 TI - Quality-of-life outcomes with laparoscopic vs open cholecystectomy. AB - BACKGROUND: Few studies have assessed the health outcomes of patients who underwent cholecystectomy. The goal of this study was to evaluate the health related quality-of-life (HRQoL) improvement of patients undergoing laparoscopic versus open cholecystectomy. METHODS: A prospective observational study was performed of consecutive patients on waiting lists to undergo cholecystectomy for nonmalignant disease in six hospitals. Patients were asked to complete two questionnaires that measure (HRQoL)-the SF-36 and the Gastrointestinal Quality of Life Index (GIQLI)-before the intervention and 3 months later. RESULTS: Improvement after surgery, measured by the SF-36 and GIQLI, was similar for both surgical techniques. The SF-health transition item showed a perception of worse health, compared to 1 year previously, for those who underwent open surgery and complications were also higher. CONCLUSIONS: HRQoL improvement at 3 months was relevant and similar for both surgical techniques, although the health transition perception was worse for those who underwent open surgery. PMID- 12730729 TI - The laparoscopic reoperation of failed Heller myotomy. AB - BACKGROUND: Laparoscopic Heller myotomy for achalasia has a 10-20% failure rate and may require reoperation to control persistent, or recurrent symptoms of dysphagia. We report our experience with laparoscopic reoperation for failed Heller myotomy. METHODS: Between 1996 and 2001, 5 patients underwent reoperative laparoscopic Heller myotomy. The mean age was 39 years. The presenting symptoms were persistent dysphagia ( n = 3), recurrent dysphagia ( n = 1), and weight loss ( n = 1). The mean duration between 1st surgery and recurrence of symptoms was 2 months and the mean duration between surgeries was 27.5 months. All operations were completed laparoscopically. RESULTS: There were no intraoperative or postoperative complications. Incomplete gastric myotomy was the cause of all 5 primary surgical failures. The mean hospital stay was 2 days. Mean follow-up was 12.8 months. Results were excellent in 2 patients who reported no dysphagia postoperatively (dysphagia grade 0) and good in 3 patients (60%) who all reported improvement in swallowing (dysphagia grade I-II). CONCLUSION: Laparoscopic reoperation for failed Heller myotomy is feasible with encouraging preliminary results. PMID- 12730731 TI - Individual bone cement spacers (IBCS) for septic hip revision-preliminary report. AB - BACKGROUND: Two-stage septic hip revision with intermediate resection arthroplasty leads to temporary poor function and difficult reimplantation. Antibiotic-loaded cement spacers improve patient mobility and maintain stability of the joint as well as length of the limb. Prefabricated spacers are readily available but lack adaptability. We developed modular moulds separately for stem and head to create more individual spacers with optimised femoral press-fit and smooth head surface. METHODS: Moulds are fabricated preoperatively by forming 4 mm polyethylene over trial stems and cups under vacuum (MS 30, Centerpulse, Switzerland). After gas sterilization, Refobacin Palacos (Merck, Darmstadt, Germany) is poured into stem and head moulds of appropriate size. A double K-wire is inserted as a rod. Fluoroscopy is used to check for air inclusions, which might impair resistance. For mechanical resistance testing trials, 5 spacers were resin-fixed with 80% stem insertion and loaded in a craniocaudal direction at 20 N/s (universal testing machine, Zwick, Germany) until failure. RESULTS: The static testing showed an average failure load of 1550 N (range 1350-2000 N). The K-wires prevented complete dislocation of the fragments. CONCLUSION: Partial weight-bearing is possible with this system, but a fall might lead to an acute fracture. New materials for the core can be tested with our biomechanical model. Following the presented craniocaudal static testing, torsion testing and dynamic testing in Ringer's solution are planned. PMID- 12730732 TI - A radiographic comparative study of two series of skeletally mature clubfeet treated by two different protocols. AB - OBJECTIVE: To compare the radiographic features of two series of congenital clubfeet to determine whether a different treatment protocol may influence the radiographic results at the end of skeletal growth. DESIGN AND PATIENTS: Two series of patients with congenital clubfeet, treated by two different manipulative techniques and by two different complementary soft tissue release operations, were radiographically studied at skeletal maturity. Twenty-one normal feet of the unilateral cases in both series served as controls. Anteroposterior and lateral radiographs of the feet were taken with the patient standing, and several radiographic parameters were studied. RESULTS AND CONCLUSIONS: The size of the talus and calcaneus and the height of the talar trochlea were smaller than normal in all cases of clubfeet, were similar in both series and were not influenced by treatment, whereas all the other radiographic parameters studied were more or less different between the two series and seemed to be influenced by treatment. In no treated clubfoot of either series was a normal radiographic foot anatomy restored, not even in those feet that had an excellent clinical result. PMID- 12730733 TI - Solitary cysticercosis of the biceps brachii in a vegetarian: a rare and unusual pseudotumor. AB - We report a 40-year-old man with cysticercosis presenting as a solitary tumor in the biceps brachii muscle. Physical examination revealed an intramuscular mass and magnetic resonance imaging suggested a cyst. The histologic diagnosis was a cysticercus. Such solitary presentation of muscular cysticercosis is extremely rare with only a handful of sporadic reports in the literature. PMID- 12730734 TI - Risk factors for hip fracture in skilled nursing facilities: who should be evaluated? AB - This paper aims to identify risk factors for hip fracture in Medicare skilled nursing facility (SNF) residents and to develop a predictive model based on routinely collected administrative data (the Minimum Data Set, MDS) to identify high-risk residents. Prospective cohort study of 28,807 North Carolina Medicare SNF residents aged >65 years with a complete MDS assessment in 1999. Demographic, historical, physical, cognitive, behavioral, activities of daily living, and medication variables were obtained from the MDS. Hip fracture occurring after the first MDS assessment identified by ICD-9 code was the outcome measure. Variables significantly associated with hip fracture by chi-square test in a randomly selected derivation sample were combined in a multivariable logistic model and in models stratified by gender. The models were validated in the remaining subjects. Variables significantly related to subsequent hip fracture in the full cohort include: female sex (odds ratio 1.3, 95% confidence interval 1.0-1.7), white race (2.3, 1.6-3.5), age (1.03 per year, 1.01-1.04), cognitive impairment (1.4, 1.8 1.8), incontinence (0.68, 0.5-0.9), prior fractures (1.6, 1.2-2.1), and prior falls (1.4, 1.2-1.8). In ambulatory non-Hispanic white women, anxiety (1.5, 1.0 2.1), anxiolytic use (1.4, 1.1-1.9), wandering (1.4, 1.0-2.2), and training in community skills (1.4, 1.1-1.8) were new significant variables. For ambulatory non-Hispanic white men, education level (2.0, 1.2-3.2), weight loss (0.5, 0.2 1.0), history of osteoporosis (3.0, 1.3-6.7), pathologic bone fracture (9.7, 2.2 42.6), COPD (2.1, 1.3-3.5), glaucoma (2.6, 1.0-6.2), and standing balance impairment (1.8, 1.0-3.3) were also significant. All models were highly correlated with subsequent hip fracture, but the discriminative ability was limited (c statistic 0.678). Risk factors explained more of hip fracture risk in non-Hispanic white men (c statistic 0.793) than non-Hispanic white women (0.658). Risk factors for hip fracture in Medicare SNF residents have similarities and differences from those previously identified in community-dwelling older adults. Osteoporosis screening and intervention should focus on the healthiest, most independent subset of residents who have the greatest fracture risk. PMID- 12730735 TI - The Amsterdam Hip Protector Study: compliance and determinants of compliance. AB - Hip protectors appear to be effective in reducing the incidence of hip fractures. However, compliance is often poor. Therefore, the objective of this study was to examine the compliance and determinants of compliance with external hip protectors. A prospective study was performed in residents from apartment houses for the elderly, homes for the elderly and nursing homes with a high risk for hip fracture (n=276). The study was performed within the framework of the Amsterdam Hip Protector Study, a randomized controlled trial examining the effect of external hip protectors on the incidence of hip fractures. Compliance was assessed by unannounced visits at 1, 6 and 12 months after inclusion in the study. During the visits, a member of the research team checked whether the participant was wearing the hip protector and, if so, whether it was worn correctly. Furthermore, data on potential determinants of compliance were collected by interviewing the participants or their nurses. Compliance was 60.8% after 1 month (n=217), 44.7% after 6 months (n=246), and 37.0% after 12 months (n=230). Of those wearing the hip protector, 86.7%, 91.7% and 96.5% of the participants were wearing the hip protector correctly after 1, 6 and 12 months respectively; and 14.8%, 16.1% and 8.8% respectively reported wearing the hip protector at night. Compliance after 12 months was predicted by the compliance after 1 month (RR=2.04; 90% CI: 1.05-3.96). Furthermore, people who experienced one or more falls in the half year before baseline had a lower probability of being compliant at 6 months (RR=0.72; 90% CI: 0.52-0.99). In conclusion, compliance is a very important issue in hip protector research and implementation. Although, the compliance percentages were moderately high during the unannounced visits in this study, not everyone was wearing the protector correctly and most participants did not wear the hip protector during the night. PMID- 12730736 TI - The true results of the ECHODIAH study with diacerein. PMID- 12730737 TI - Predicting the failure load of the distal radius. AB - The distal radius is an important site for the early detection of patients at risk for fracture. Since measuring bone strength in vivo is not possible, we evaluated which bone assessment method of the forearm would best predict failure load of the distal radius and computed a factor of risk for wrist fracture (Phi wrist). Thirty-eight cadaveric forearm specimens were measured by five different techniques to assess bone density, bone mineral content, geometry and trabecular structure at the distal forearm. The bone assessment techniques included dual energy X-ray absorptiometry (DXA) of the radius, peripheral quantitative computed tomography (pQCT) of the 4% and 20% distal sites of the radius, DXA of the phalanges, digital X-ray radiogrammetry of the forearm (DXR-BMD), and quantitative ultrasound of the radius. The failure load of each excised radius was determined by simulating a fall on an outstretched hand. The pQCT measurements of polar stress-strain index and cortical content explained the greatest portion of variance in failure load (r2=0.82-0.85). Bone mineral content measures were generally better predictors of failure load (r2=0.53-0.85) than the corresponding volumetric or areal bone mineral density values (r2=0.22-0.69) measured by either pQCT or DXA. Multiple regression analysis showed that the addition of a bone geometry measure improved the ability of a bone density measure alone to predict failure load. There was high variability in the ability of different techniques and different variables within a given technique to predict failure load. Estimates of the factor of risk for wrist fracture (Phi wrist) revealed that the women in this study would have been likely to fracture their distal radius upon falling from a standing height (Phi wrist= 1.04), whereas the men would have likely withstood the impact without fracturing their wrist (Phi wrist= 0.79). PMID- 12730738 TI - The diagnostic role of dual femur bone density measurement in low-impact fractures. AB - A high correlation has been documented between the left and right femoral bone mineral densities in the normal population. This suggests that dual femur measurements are not justified in clinical practice. This study evaluated whether this premise holds for subjects who have lost bone mass and have sustained fractures with minimal trauma. Seventy-eight women aged 31-83 years (mean=66 years) with previous low-impact fractures had both proximal femora measured using dual-energy X-ray absorptiometry. There were significant correlations between values in the left and right total hip (TH) (r=0.95; p<0.05) and in the left and right femoral neck (FN) (r=0.90; p<0.05). The mean differences between the left and right TH and FN densities were not significant. However, the range of the limits of agreement for the TH (-0.074 to 0.086 g/cm2) and FN (-0.115 to 0.105 g/cm2) were greater than the 95% confidence interval for true change for the TH (0.05 g/cm2) and FN (0.07 g/cm2). Any longitudinal BMD assessment therefore needs to measure the same proximal femur to get a reliable comparison. A one-tailed analysis showed that for the TH, 7.5% of subjects had a T-score discordance greater than or equal to 0.5 and 0.5% had a T-score discordance greater than or equal to 1. For the FN, 9% had a T-score discordance greater than or equal to 0.5 and 2.5% had a T-score discordance greater than or equal to 1. The use of dual femur measurements increases the diagnostic yield by about 10% in subjects with prior minimal trauma fractures. PMID- 12730739 TI - The impact of a simple fracture clinic intervention in improving the diagnosis and treatment of osteoporosis in fragility fracture patients. AB - We examined the effect of a fracture clinic intervention in reducing previously documented undertreatment of osteoporosis (OP) in individuals with fragility fractures. Fragility fracture patients presenting to five community fracture clinics with no prior diagnosis of, or treatment for OP, and whose radiographic appearance was consistent with fragility fracture, were included. These individuals (intervention group) were informed of their OP risk, and advised to follow up with their physician for assessment. A standardized letter, intended for the physician and outlining the same was provided. Three months later, a telephone interview determined whether a physician visit had occurred, and if so, what investigation and treatment recommendations were made. These outcomes were compared with those for an equal number of age- and sex-matched fragility fracture "controls," selected from among fracture clinic attendees in the 6-9 months preceding the intervention. Logistic regression was used to examine the effect of having received the intervention on physician follow-up, bone density testing, and OP treatment recommendations. The mean age of the 278 participants (139 per group) was 66.0 years; 74% were female. Adjusting for age, sex, hospital, and perceived diagnosis of OP, those who received the intervention were more likely to follow up with a physician (adjusted OR 1.85, p=0.02) and to be recommended bone density testing (adjusted OR 5.22, p<0.0001), but were not more likely to receive an OP treatment recommendation (adjusted OR 2.07, p=0.07). It is concluded that a simple fracture clinic intervention increased follow-up and investigation, but not treatment for OP, in fragility fracture patients. Individuals recommended treatment for OP were more likely to perceive themselves as having OP and to have had a previous fragility fracture. Our findings suggest that future interventions should incorporate assessment of patients' OP health beliefs and education about risk factors for fracture, and should be coupled with physician education to achieve optimal results. PMID- 12730741 TI - Contribution of clinical risk factors to bone density-based absolute fracture risk assessment in postmenopausal women. AB - Hip fractures are independently associated with advancing age, specific clinical risk factors (CRFs), and low bone mineral density (BMD). The use of BMD T-scores for quantifying fracture risk ignores the contribution of age and CRFs. We previously developed a mathematical model of absolute hip fracture risk that incorporates patient age, BMD, and the results of eleven specific CRFs. The purpose of this study was to compare the contribution of an approach to fracture risk stratification using the full model (age, CRFs and BMD) with that of a unidimensional BMD-only model. We selected 213 consecutive postmenopausal females (mean age 65.3, range 50-87.9) with CRF data referred for BMD assessment of fracture risk. Absolute hip fracture risk (over the next 5 years and remaining lifetime) was estimated using both the full and BMD-only models. The mean ratio of absolute hip fracture risks (BMD-only/full model) derived for each patient was 0.8 (95% CI, 0.16-4.0) for hip fracture in the next 5 years and 1.1 (CI, 0.1-7.6) for remaining lifetime. The wide confidence intervals indicate a large contribution of age and CRFs to fracture risk stratification. Categorization of women as "high risk" was frequently discordant for the two models. One-half of the women designated "high risk" under the full model were classified as "low risk" based upon BMD alone. In conclusion, we have shown that a multidimensional approach to hip fracture risk stratification is feasible, and greatly modifies risk stratification based on BMD alone. PMID- 12730740 TI - Changes in bone mineral density in the hip and spine before, during, and after the menopause in elite runners. AB - This study was undertaken to determine the degree of interaction between hormonal and athletic effects on changes in bone mineral density (BMD, g/cm(2)) with time in women around the menopause. Thirty-five elite female runners aged over 40 participated in a longitudinal cohort study of hip and spine bone density changes and had two measurements a mean of 4.1 years apart. Eighteen remained estrogenized throughout the study (E+ to +), ten remained postmenopausal throughout (E- to -), while seven showed a change in status (E- to +) or (E+ to ). At study exit, both postmenopausal E+ and E- subjects had significantly higher than expected broadband ultrasound attenuation (BUA) and velocity of sound (VOS) values at the calcaneus. However, Z-score 95% confidence intervals for BMD of the hip and spine embraced zero except at the femoral trochanter in E+ women. The E+ to + group did not lose bone at any site. The E- to - group lost bone significantly, but only at the femoral neck and spine at rates close to 1% annually. Multiple analysis of variance repeated measures analysis showed a significant interaction in the effects of estrogen exposure with DXA measurement site, with the femoral trochanter being less affected by E- status than the spine and femoral neck. Body mass index (BMI: weight/height(2), kg/m(2)) had a significantly positive effect on total hip and femoral neck BMD independently of estrogen status. In conclusion, in this pilot study, postmenopausal elite runners not on hormone replacement therapy lost BMD from the femoral neck and spine at the expected rates but in the femoral trochanter bone loss appeared reduced and in the calcaneus BUA and VOS were maintained close to young normal values. PMID- 12730744 TI - The effect of DXA scanning on clinical decision making by general practitioners: a randomized, prospective trial of direct access versus referral to a hospital consultant. AB - The objective of the study was to assess the impact of direct access DXA scanning (DADS) upon GPs' management decisions in patients considered to be at risk of osteoporosis. It was designed as a randomized, prospective, parallel group trial, set within the primary care environment and a university teaching hospital. The participants were 330 patients aged 31 to 89 years from 18 general practices in Edinburgh. Patients were randomized to either DADS or to the current system of specialist referral (controls). The primary outcome measure was frequency of change of management after DXA scanning. Secondary outcome measures were: change in health status, adherence to therapy, clinical events and resource use at one year follow-up. The primary outcome was that 60% each of DADS patients (98/165) and controls (99/165) had changes in management following DXA scanning. In 30% of patients (12/41) in whom GPs had proposed changing management even in the absence of a scan, different therapy was chosen after the scan (no difference between DADS and control groups). There was an improvement in health utility (p =0.014 for both groups combined), differing slightly between the two groups even after age correction (p =0.014). 68% of the DADS group and 70% of controls were adherent to therapy after one year. In terms of clinical events, at one year there was one major adverse event (control group patient), 5 new fractures in the DADS group and 3 in controls - there were no hip fractures in this study. With regard to resource use, there were 24 referrals to hospital specialist after DXA scanning among the DADS group, vs 12 among controls (p < 0.05). The total number of visits to health professionals was 525 in DADS and 585 in controls (p=ns); mean waiting time from randomization to receipt of report/clinic letter was 4 weeks for DADS vs 13 weeks for controls( p < 0.0001). In conclusion, DXA scanning resulted in management change in at least 60% of cases. Direct access does not result in a clinical outcome significantly different from a consultant led service, and is more economically efficient than the current model of hospital referral. PMID- 12730745 TI - Tolerability and compliance with risedronate in clinical practice. AB - The purpose of this study was to determine compliance with dosing instructions, and the prevalence of possible adverse events, when risedronate is used in clinical practice. 219 patients were studied. We found that despite counseling one in four patients were non-compliant with dosing instructions. Those patients who did not stay upright after taking risedronate were more likely to have an adverse event and to discontinue the drug. Adverse events were experienced by 38% of patients, the commonest being gastrointestinal. Upper GI adverse events occurred in 21% of patients. A previous history of upper GI symptoms applied to 44% of patients and significantly more of them experienced upper GI adverse events than those with no history of GI problems. Forty two (19%) of patients taking risedronate stopped therapy due to adverse events, but only ten of these patients had contacted the Osteoporosis Unit about these symptoms. Approximately one third of the patients who experienced adverse events in this study had attempted a rechallenge with the drug. This was worthwhile, however, as in almost 50% of these patients their symptoms settled and they continued with therapy. This study has highlighted the importance of following up patients on long-term osteoporosis medication to ensure optimal compliance. The use of specialized osteoporosis nurses in clinics or primary care to follow up patients needs to be addressed. PMID- 12730743 TI - Isoflavones and skeletal health: are these molecules ready for clinical application? AB - A review of the recent literature on the effects of isoflavones was undertaken to determine whether molecules such as genistein and daidzein, aglycone derivatives of soybeans, might have benefit in the prevention and treatment of osteoporosis. The current standard for science-based medicine is the documentation of efficacy of an agent under controlled, randomized, prospective conditions. A few short clinical investigations have been undertaken using isoflavones (along with soy protein), but they had limitations in study design, and the numbers of women studied were small. Other evidence from animal models, in vitro experiments, and epidemiological reports suggest that the isoflavones have skeletal benefits in women with little or no ovarian estrogen production. A clear need exists for prospective human trials, using the required conditions of randomized clinical trials and designs, to satisfy objectively the needs for science-based medicine and for appropriate clinical applications. PMID- 12730747 TI - Hip axis length in mid-life Japanese and Caucasian U.S. residents: no evidence for an ethnic difference. AB - The rate of hip fracture in Japanese women is approximately 50% lower than that of Caucasian women. Shorter hip axis length (HAL) in Japanese women is one possible explanation underlying the observed ethnic difference in fracture risk. We measured HAL in a community-based sample of Japanese (n=267) and Caucasian (n=199) women, aged 46 years on average, residing in Los Angeles, California. We hypothesized that HAL would be shorter in the Japanese women compared with the Caucasian women, and that this ethnic difference would persist after height adjustment. Unadjusted mean HAL was 101.45 mm in Japanese women and 105.98 mm in Caucasians (p<0.0001). Average height-adjusted HAL was 103.24 and 103.58 mm in Japanese and Caucasians, respectively (p=0.52). The relation between HAL and height was linear across the range of values observed in each ethnicity. Results were similar in analyses that compared Caucasians to Japanese women born in the US (n=142) or outside the US (n=125). We conclude that there was no Japanese Caucasian difference in height-adjusted HAL in our study sample. PMID- 12730742 TI - Gene expression studies of osteoporosis: implications for microarray research. AB - Osteoporosis is one of the most common bone-related disorders in the elderly. It is a complex disease, and largely determined genetically. Traditional gene expression studies have shown that osteoporosis has complex regulating mechanisms which are controlled by multiple inherent factors, such as hormones, cytokines, various receptors, etc. The complex nature of osteoporosis, and the large number of genes involved in its onset and development, suggest the use of the state of the art microarray technique as a powerful tool to unravel mechanisms underlying etiology of osteoporosis. PMID- 12730746 TI - A double-blind dose-ranging study of risedronate in Japanese patients with osteoporosis (a study by the Risedronate Late Phase II Research Group). AB - To determine the clinical recommended dosage regimen of risedronate for the treatment of involutional osteoporosis in Japanese patients, dose-response relationships for the efficacy and safety of this drug were investigated using a multi-center, randomized, double-blind, parallel group comparative design with four dose levels of risedronate (placebo, 1 mg, 2.5 mg and 5 mg per day). A total of 211 patients diagnosed with involutional osteoporosis according to the criteria proposed by the Japanese Society for Bone and Mineral Research were randomized and received one of the four doses once daily for 36 weeks. All patients were supplemented with 200 mg of calcium daily in the form of calcium lactate. The primary efficacy endpoint was the percent change in bone mineral density of the lumbar spine (L2-L4 BMD) determined by dual-energy X-ray absorptiometry (DXA) from baseline to the time of final evaluation. Changes in biochemical markers of bone turnover and safety profile were also compared. Percent changes in L2-L4 BMD at final evaluation in the placebo, and 1-, 2.5-, and 5-mg risedronate groups were 0.79+/-5.30, 2.71+/-4.93, 5.29+/-3.96, and 5.15+/-4.25% (mean+/-SD), respectively. A linear dose-response relationship was obtained up to a dose of 2.5 mg, whereas no further increase in BMD was observed at 5 mg. The decrease in bone turnover markers, including N-terminal osteocalcin, phosphorus, and urinary deoxypyridinoline, also showed a linear dose-response relationship up to a dose of 2.5 mg. Alkaline phosphatase level decreased linearly up to a dose of 5 mg. Risedronate was well tolerated in this 36-week study with 1- to 5-mg doses. Neither the overall incidence of adverse events nor the percentage of patients without problem in overall safety assessment differed significantly among the dose groups including the placebo group. Based on these results, a once-daily dose of 2.5 mg of risedronate, which is half that used in Caucasians, is recommended for the treatment of involutional osteoporosis in Japanese patients. PMID- 12730748 TI - Should the treatment of osteoporosis be more selective? PMID- 12730749 TI - Comparison of ultrasound and X-ray absorptiometry bone measurements in a case control study of female rheumatoid arthritis patients and randomly selected subjects in the population. AB - To compare quantitative ultrasound (QUS) and dual-energy X-ray absorptiometry (DXA) bone measurements in female rheumatoid arthritis (RA) patients and controls were randomly selected from the population; secondly, to examine disease and demographic factors associated with these bone measurements. In a total of 115 RA patients (mean age 63.0 years) and 115 age- and gender-matched controls demographic and clinical variables were collected and heel QUS parameters [speed of sound (SOS), broadband ultrasound attenuation (BUA) and stiffness index (SI)] as well as DXA bone mineral density (BMD) at spine and hip were measured. The differences in QUS and DXA measurements between RA patients and controls were tested both on a group and on an individual level. Univariate and multivariate statistical tests were applied to explore for associations to the bone measurements. In the RA patients mean disease duration was 16.6 years, erythrocyte sedimentation rate 23.6 mm/h, M-HAQ 1.68, 28-swollen joint count 7.7, 18-deformed joint count 4.5, 50.0% were rheumatoid factor (RF) positive and 44.2% were current users of prednisolone. All bone measurements were reduced in RA patients compared with controls (SOS 1.9%, BUA 9.4%, SI 19.5%, femoral neck BMD 7.4%, total hip BMD 7.5%, spine L2-L4 BMD -3.0%). Only at spine was the BMD reduction not statistically significant ( P=0.21). In the subgroup of never users of prednisolone SOS was decreased by 1.4%, BUA by 3.7%, SI by 11.0, femoral neck BMD by 2.7%, and total hip BMD by 0.6%, whereas for spine L2-L4 BMD was increased by 4.3% and only for SOS and SI was the decrease statistically significant. The QUS discriminated better than DXA between patients and controls on a group level, but this difference in favor of QUS disappeared on an individual level when the measurement errors were taken into account. Age, BMI, RF and deformed joint count, but not corticosteroids, were independently associated with at least one of the QUS and one of the DXA measures; however, the association between disease related variables was stronger with the QUS bone measures than with the DXA bone measures. The results for the quantitative QUS bone measures seem to mainly reflect bone mass. Disease-related variables in multivariate analysis remained independently associated with all QUS measures even when adjusting for DXA bone measures. Further studies are needed to examine if QUS may reflect other aspects than bone mass and be a potential better predictor for fracture risk in RA and corticosteroid-induced osteoporosis. PMID- 12730750 TI - Cost-equivalence of different osteoporotic fractures. AB - METHODS: Among 985 Olmsted County, Minnesota, residents who experienced an osteoporotic fracture (distal forearm, humerus, clavicle/scapula/sternum, ribs, vertebrae, pelvis, hip, other femur or tibia/fibula [the latter in women only]), we estimated the incremental cost of direct medical care in the following year compared with age- and sex-matched controls without a fracture randomly sampled from the same community. RESULTS: The overall median incremental (case minus control) cost in the succeeding year was $2,390, with a particularly high incremental cost for hip fractures ($11,241). There was fair concordance between the incremental cost of the different fractures, relative to hip fracture alone, and the previously published disutility associated with each fracture type relative to hip fracture. Overall, the incremental cost for all osteoporotic fractures combined was 46% greater than that for hip fractures alone in women and 47% greater in men. This is consistent with the earlier report that overall morbidity from all osteoporotic fractures combined is 47% and 39% greater in women and men, respectively, than the morbidity attributable solely to hip fractures. CONCLUSION: These data lend support to the notion that other osteoporotic fractures can be quantified relative to hip fracture on the basis of their cost, as well as their morbidity and mortality. This may simplify health economic analyses by allowing all fracture outcomes to be modeled relative to hip fractures (i.e., hip fracture 'equivalents') and will provide a more comprehensive assessment of osteoporosis outcomes than is possible by focusing only on hip fractures. PMID- 12730751 TI - Early menopause: increased fracture risk at older age. AB - The aim of this study was primarily to determine the relationship between early menopause and the presence of fractures later in life, and secondly, to check for the significance of confounding factors (such as smoking habits, body mass index (BMI), weight and use of hormones). In this cross-sectional population based study, the subjects were 4725 postmenopausal women, 50-80 years of age, registered with 23 general practitioners (GPs). For the purpose of the present study, the total population was analyzed as well as the subgroup of 2757 women (the study population) with a natural menopause. Medical history questionnaire, weight, height and bone mineral density measurements were taken. Bivariate and multivariate analyses were carried out with documented fractures in three categories: during lifetime; after menopause and after age 50 years) as dependent variable and age, BMI, bone mineral density, weight, smoking habits, use of hormones and early menopause as independent variables. The total study population as well as the subgroups "early" and "normal menopause", stratified in three 10 year and in six 5-year categories, were analyzed. Results are expressed as odds ratio and 95% confidence intervals (CI). Multivariate logistic regression analysis revealed that over 70 years of age, BMD< or =0.800 ( t-score<2.5) and early menopause were the only systemic independent predictors of all three fracture categories. Comparing the subgroups normal menopause and early menopause, the early menopause group showed a statistically significant higher overall fracture rate (OR=1.5; CI 1.2-1.8). Over age 70, the difference in the prevalence of fractures reached statistical significance in each age category (OR: 1.8 and 2.1, respectively). Smoking was found to be associated with early menopause (OR=1.5; CI 1.2-1.8) but not with the presence of fractures. Height above 165 cm was found to be associated with a higher prevalence of fractures during lifetime. The present study shows that early menopause is statistically significant associated with the presence of fractures during lifetime, after age 50 years and after menopause. Especially at older age, early menopause is an important predictor of fractures. PMID- 12730752 TI - Phalangeal quantitative ultrasound technology and dual energy X-ray densitometry in patients with primary hyperparathyroidism: influence of sex and menopausal status. AB - Fifty-one patients with surgically proven primary hyperparathyroidism (PHPT), 11 males and 40 females, mean age+/-SD: 55.9+/-14.1 years, and 58 age- and sex matched normal subjects were studied. The femoral and L(2)-L(4) bone mineral density (BMD; Hologic QDR 4500 C), as well as quantitative ultrasonometry (QUS; DBM-Sonic 1200) of the phalanges of both hands were measured in patients and controls. QUS measurements included amplitude-dependent speed of sound (AD-SoS), and other parameters derived from the graphic trace: signal dynamics (Sdy), first wave amplitude (FWA), bone transmission time (BTT) and ultrasound bone profile index (UBPI). Patients with PHPT showed significantly lower dual energy X-ray densitometry (DXA) values and QUS parameters compared to controls (lumbar spine Z score: controls: -0.16+/-1.12, PHPT: -0.70+/-1.14, P=0.016; femoral neck Z-score: controls: -0.28+/-1.74, PHPT: -1+/-1.01, P=0.013; total femur Z-score: controls: 0.33+/-1.12, PHPT: -1.01+/-0.95, P=0.0013; AD-SoS Z-score: controls: -0.89+/ 1.22, PHPT: -1.97+/-1.78, P=0.0003; FWA Z-score: controls: 0.36+/-1, PHPT: 0.62+/ 0.85, P<0.0001; BTT Z-score: controls: 0.04+/-1.03, PHPT: -0.45+/-1.37, P=0.044; UBPI Z-score: controls: -0.02+/-1.01, PHPT: -0.68+/-1.05, P=0.002; SDy (mV/micros(2)): controls: -295+/-256, PHPT: -498+/-306, P=0.0003). In male patients, BMD values measured on the lumbar spine and femoral regions were similar to those found in male controls, while QUS values were significantly lower (lumbar spine Z-score: controls: -1.05+/-1.41, PHPT: -1.75+/-1.21, P=0.21; femoral neck Z-score: controls: -0.37+/-1.84, PHPT: -1.11+/-1.14, P=0.27; total femur Z-score: controls: -0.16+/-1.59, PHPT: -1.02+/-1.20, P=0.168; AD-SoS Z score: controls: -0.52+/-1.58, PHPT: -1.57+/-1.77, P=0.149; FWA Z-score: controls: 0.67+/-1.01, PHPT: -0.74+/-0.79, P=0.0016; BTT Z-score: controls: 1.22+/-0.83, PHPT: 0.75+/-1.51, P=0.478; UBPI Z-score: controls: 0.56+/-0.94, PHPT: -0.47+/-1.10, P=0.025; SDy (mV/micros(2)): controls: -167+/-230, PHPT: 485+/-307, P=0.01). Women with PHPT were further divided into two subgroups: premenopause ( n=11) and postmenopause ( n=29). The premenopausal women with PHPT showed significantly lower DXA values than those of the premenopausal control ones, but similar QUS parameters (lumbar spine Z-score: controls: 0.12+/-0.66, PHPT: -0.59+/-0.85, P=0.03; femoral neck Z-score: controls: 0.06+/-2.85, PHPT: 1.48+/-1.05, P=0.11; total femur Z-score: controls: -0.51+/-0.97, PHPT: -1.48+/ 0.63, P=0.009; AD-SoS Z-score: controls: 0.78+/-0.89, PHPT: -1.26+/-1.88, P=0.42; FWA Z-score: controls: 1.14+/-0.77, PHPT: 0.12+/-0.80, P=0.007; BTT Z-score: controls: 0.13+/-0.60, PHPT: 0.25+/-1.15, P=0.757; UBPI Z-score: controls: 0.73+/ 0.49, PHPT: 0.24+/-0.96, P=0.15; SDy (mV/micros(2)): controls: -118+/-123, PHPT: 271+/-301, P=0.106). The postmenopausal women with PHPT showed both DXA and QUS parameters significantly lower than those found in the postmenopausal controls (lumbar spine Z-score: controls: 0.09+/-0.96, PHPT: -0.31+/-0.96, P=0.004; femoral neck Z-score: controls: -0.38+/-1.01, PHPT: -0.76+/-0.91, P=0.14; total femur Z-score: controls: -0.33+/-0.97, PHPT: -0.81+/-0.92, P=0.057; AD-SoS Z score: controls: -1.08+/-1.17, PHPT: -2.38+/-1.68, P=0.31; FWA Z-score: controls: -0.013+/-0.81, PHPT: -0.86+/-0.74, P=0.0009; BTT Z-score: controls: -0.58+/-0.68, PHPT: -1.13+/-0.93, P=0.016; UBPI Z-score: controls: -0.62+/-0.83, PHPT: -1.11+/ 0.82, P=0.034; SDy (mV/micros(2)): controls: -419+/-242, PHPT: -589+/-269, P=0.012). The relative risk of osteopenia was significantly increased in PHPT patients at several measurement sites. There was a highly significant correlation between spine and femoral BMD and QUS parameters, while PTH serum levels did not correlate with any of the densitometric variables. In conclusion, QUS parameters would seem to be able to distinguish patients with PHPT from normal controls in male subjects and in postmenopausal women, but not in premenopausal women. This would suggest that the higher estrogen levels in premenopausal patients might preserve the bone from significant structural changes. This may also suggest that hyperparathyroidism, in addition to the reduction of bone mineral content, can cause an alteration of bone structure with an additional increase in fracture risk in postmenopausal women. Furthermore, the alterations in QUS parameters in patients who do not show significant changes in DXA measurements suggest an involvement of bone that is independent of mineral content and may be helpful for selecting candidates for surgery, according to NIH criteria. PMID- 12730753 TI - Pubertal bone growth in the femoral neck is predominantly characterized by increased bone size and not by increased bone density--a 4-year longitudinal study. AB - Fragility fractures are correlated to reduced bone size and/or reduced volumetric bone density (vBMD). These region-specific deficits may originate from reduced mineral accrual and/or reduced skeletal growth during the first 2 decades of life. Before pathological development can be defined, normal skeletal growth must be described. To evaluate growth of bone size, accrual of bone mineral content (BMC), areal bone mineral density (aBMD) and vBMD in a population-based cohort, 44 boys and 42 girls were followed by annual measurements from the age of 12 to 16 (attendance rates 90-100%). Segmental bone length, bone width, BMC, aBMD and vBMD were measured by dual-energy X-ray absorptiometry (DXA). Data were compared with predicted adult peak, as determined in 36 men aged 27.7+/-4.6 years and 44 women aged 26.8+/-4.9 years. Growth in width of the femoral neck precedes accrual of BMC in the femoral neck in both genders up to age 15. The girls were at all ages closer to their predicted adult peak in both bone width and BMC compared with the boys except in the femoral neck. As femoral neck vBMD had reached its predicted adult peak already at 12 years in both genders, the increase in femoral neck BMC and femoral neck aBMD from age 12 to 16 was most likely to be explained by the increase in bone size. In boys the peak velocity growth was recorded at ~14 years for BMC, height, width and lean mass. Growth from the age of 12 to 16 seems to build a bigger but not a denser skeleton in the femoral neck. PMID- 12730754 TI - The effect of vertebral fracture as a risk factor for osteoporotic fracture and mortality in a Spanish population. AB - There is little data concerning the morbidity, mortality, and epidemiology of vertebral fracture. The aim of this study was to evaluate the effect of prevalent and incident vertebral fractures as risk factors for further osteoporotic fractures and mortality. The study was performed on a cohort of 316 women and 308 men older than 50 belonging to the EVOS study, randomly selected from our city register. At the beginning of the study and 4 years later, lateral dorsal and lumbar X-rays were performed. In addition, evaluation of the incidence of osteoporotic nonvertebral fractures was performed throughout 8 years. The incidence of all osteoporotic fractures was higher in women than in men (two-fold increase in vertebral fracture incidence and five-fold increase in Colles' and femur incidence). Vertebral fracture was a strong risk factor for a new vertebral fracture [RR=4.7 (1.8-11.9)], hip fracture [RR=6.7 (2.0-22.7)] and Colles' fracture [RR=3.0 (1.1-7.8)]. Prevalent and incident vertebral fractures were associated with a higher risk of having a hip fracture [RR=10.0 (2.0-50.2)] and Colles' fracture [RR=5.5 (1.3-23.4)]. In addition, in women, the vertebral fracture was associated with a higher mortality. By contrast, no association was found in men. These results demonstrate the association between a previous vertebral fracture with increments in the incidence of osteoporotic fractures of any type. In addition, we found a significantly higher mortality rate in women having vertebral fractures. These findings support the necessity of preventing the occurrence of vertebral fractures to limit their strong negative impact on mortality. PMID- 12730755 TI - Place of residence and risk of fracture in older people: a population-based study of over 65-year-olds in Cardiff. AB - Fracture prevention strategies will be most cost-effective if targeted on groups of frail elderly people who are at particularly high risk of falls and fractures. Elderly people living in care homes are one potential target population, but fracture incidence in this setting remains poorly defined in many countries. We used the All Wales Injury Surveillance System (AWISS) in a population-based study of people aged over 65 years living in the city of Cardiff. We linked a postcode based register of all sheltered accommodation and all residential and nursing homes in the city with injury data from Cardiff's single Accident and Emergency Department. Cardiff has 47,700 residents aged over 65 years with 1918 (4.0%) living in residential or nursing homes and 1868 (3.9%) in sheltered accommodation. In 1999 we identified a total of 1305 fractures including 366 hip fractures, giving a crude fracture incidence of 27.4/1000 per year and 7.7/1000 per year, respectively. Care home residents suffered 213 fractures, 95 of which were of the hip, giving a crude fracture incidence of 111.1/1000 per year and 49.5/1000 per year, respectively. People living in sheltered accommodation suffered 94 fractures, including 28 at the hip, giving a crude fracture incidence of 51.4/1000 per year and 15.3/1000 per year, respectively. People in care homes and sheltered accommodation tend to be older than those living in the community, and we adjusted for this by calculating age- and gender-standardized relative ratios for each setting. Compared with the community dwelling population, care home residents had an overall fracture risk of 2.9 (95% CI 2.5-3.3) and a hip fracture risk of 3.3 (95% CI 2.6-4.2). People in sheltered accommodation had a total fracture risk of 1.7 (95% CI 1.4-2.1), and a hip fracture risk of 1.6 (95% CI 1.1-2.4). Such figures support the potential cost-effectiveness of strategies that seek to prevent fractures in care homes and sheltered accommodation, and are of special interest to those planning intervention studies in these settings. PMID- 12730756 TI - Use of matched historical controls to evaluate the anti-fracture efficacy of once a-week risedronate. AB - Placebo controls are essential to assess anti-fracture efficacy of new osteoporosis therapies, but inclusion of a placebo arm in a subsequent clinical trial may be limited by practical or ethical considerations; in these cases, use of an historical control may be appropriate. A recent active-controlled study of risedronate 35 mg once a week demonstrated that this regimen produces increases in bone mineral density (BMD) that are comparable to those seen with the risedronate 5 mg daily dose, which has proven anti-fracture efficacy. To assess the anti-fracture efficacy of this new regimen, we have analyzed the fracture data collected in an active controlled study of risedronate dosing regimens (the Once-a-Week study) using matched historical control data from previous placebo controlled trials. Women in the Once-a-Week study were matched for age, years since menopause, BMD, and prevalent vertebral fracture status, with placebo patients in the Vertebral Efficacy of Risedronate Therapy (VERT) trials forming an historical placebo group. We also constructed an historical active treatment group from the 5 mg daily arm of the VERT trials for comparison with the 5 mg daily and 35 mg once weekly treatment groups in the Once-a-Week study. Data were obtained from the risedronate 5 mg daily group (n=480) and 35 mg once-a-week group (n=485) in the Once-a-Week study and historical control groups representing daily placebo patients (n=114, matched from 993) and risedronate 5 mg daily patients (n=120; matched from 990) in the VERT studies. Patients received calcium supplementation (1000 mg daily); vitamin D was given if baseline serum 25 hydroxyvitamin D levels were low. Over 1 year, new vertebral fracture risk in the 35 mg once-a-week group was reduced by 77% relative to the historical placebo group (1.2% versus 5.0%; RR 0.23; 95% CI, 0.54 to 0.91, P=0.018), similar to the 1-year risk reduction observed in the VERT trials of risedronate 5 mg daily (61 65%). The incidence of new vertebral fractures in the three active treatment groups was similar: 1.7% in the historical risedronate 5 mg group, 1.5% in the risedronate 5 mg daily group from the Once-a-Week study, and 1.2% in the 35 mg once-a-week group. Risedronate 35 mg once a week appears as effective as the 5 mg daily dose in reducing the risk of new vertebral fractures in the first year of treatment. The use of appropriate historical control data provides an approach to the assessment of fracture effects in osteoporosis trials for which placebo controlled data are not available. PMID- 12730757 TI - Pharmacovigilance study of alendronate in England. AB - Alendronate sodium is an aminobiphosphonate, an analog of inorganic pyrophosphate, indicated for the treatment of osteoporosis in post-menopausal women. We analyzed events reported in patients prescribed alendronate by general practitioners (GPs) in England. A non-interventional observational cohort study was conducted using the technique of prescription event monitoring (PEM). Exposure data were obtained from dispensed prescriptions issued between October 1995 and January 1997. Outcome data were obtained by sending questionnaires to prescribing GPs. The cohort comprised 11,916 patients. Events most frequently reported as suspected adverse drug reactions and reason for stopping alendronate were recognized gastrointestinal events listed in the Summary of Product Characteristics. These included nausea/vomiting, abdominal pain, dyspepsia, esophagitis and esophageal reflux. Events with the highest incidence density (ID(1) per 1000 patient months treatment) were dyspeptic conditions (32.2), nausea/vomiting (20.8) and abdominal pain (13.8). The term dyspeptic conditions included dyspepsia, esophagitis, esophageal reflux, duodenitis, gastritis and heartburn. Serious suspected adverse reactions possibly related to alendronate were single reports of angioedema, erythema multiforme, hypercalcemia and hypocalcemia. There were 540 deaths in this elderly cohort. This study suggests that alendronate appears to be well tolerated, though there may be risk of developing gastrointestinal side effects including esophagitis and esophageal ulcers. PMID- 12730758 TI - Inhaled corticosteroids effects on bone in asthmatic and COPD patients: a quantitative systematic review. AB - Deleterious effect of oral corticosteroids on bone has been well documented, whereas this remains debated for inhaled ones (ICS). Our objectives were to analyze the effects of ICS on bone mineral density, fracture risk and bone markers. We performed an exhaustive systematic research of all controlled trials potentially containing pertinent data, peer-reviewed by a dedicated WHO expert group, and comprehensive meta-analyses of the data. Inclusion criteria were ICS, and BMD/markers/fractures in asthma/chronic obstructive pulmonary diseases (COPD) and healthy patients. Analyses were performed in a conservative fashion using professional dedicated softwares and stratified by outcome, study design and ICS type. Results were expressed as standardized mean difference/effect size (ES), relative risk (RR) or odds ratio (OR), depending on study design and outcome units. Publication bias was investigated. Twenty-three trials were reviewed; 11 papers fit the inclusion criteria and were assessed for the main analysis. Quality scores for the randomized controlled trials (RCTs) were 80%, 71% for the prospective cohort studies, and 78% for the retrospective cohort and cross sectional studies. We globally assessed ICS effects on BMD and found deleterious effects: ES=0.61 ( p=0.001) for healthy subjects, and ES=0.27 ( p<0.001) for asthma/COPD patients. For these patients, this effect was 0.21 ( p<0.01) at the lumbar spine, and 0.26 ( p<0.001) at the hip or femoral neck. A single study evaluated the impact of ICS on hip fracture and reported an increased OR of 1.6 (1.24; 2.03). Lumbar fracture rate differences did not reach the level of statistical significance: 1.87 (0.5; 6.94). Osteocalcin and PICP were decreased and ICTP, pyridinoline and deoxypyridinoline levels were not significantly affected. Budesonide (BUD) appeared to be the ICS inducing the less deleterious effects on bone, followed by beclomethasone dipropionate (BDP) and triamcinolone (TRI). Publication bias investigation provided non-significant results. In our meta-analyses, BUD at a mean daily dose (SD) of 686 microg (158 microg), BDP at 703 microg (123 microg) and TRI at 1,000 microg (282 microg) were found to affect bone mineral density and markers in patients suffering from the two major respiratory diseases. These findings could have practical implication in the long term management of asthmatic and COPD patients. PMID- 12730759 TI - The hospital cost of vertebral fractures in the EU: estimates using national datasets. AB - The purpose of this study was to estimate the hospital cost of vertebral fractures in the EU using national datasets to explore some of the methodologic limitations associated with such an approach. Hospital costs for vertebral fractures across the EU were compared with the hospital costs associated with hip fractures. Additionally, these costs were placed into the health care context by making comparisons with national health care expenditure. All EU Ministries of Health were contacted to identify national datasets to estimate the average length of stay, cost per diem and the number of patients discharged with vertebral fractures. Where national information was not available expert opinion and data from the relevant literature were used instead. Countries show a marked difference in the length of stay between men and women, with differences ranging from 0.32 days in Austria to 20.2 days in Spain. The average hospitalization rate was found to be 8% across the EU, with higher rates found for men than for women. Interestingly a positive correlation between health expenditure per capita and hospitalization rates was found. The total cost of vertebral fractures in the EU was estimated at euro 377 million per year. Across the EU the hospital cost of a vertebral fracture was on average 63% that of a hip fracture. National datasets allow us to estimate the cost of vertebral fractures in the EU but show limitations. In the absence of large scale prospective studies, national datasets need to be further refined to ensure more accurate estimations of the cost of vertebral fractures in the EU. PMID- 12730760 TI - Low incidence of hip fractures in an equatorial area. AB - Hip fractures in patients older than 60 years of age represent a serious morbidity linked to osteoporosis that contributes to its mortality rates. Both genetic and environmental influences have been reported as important factors related to the epidemiology of osteoporosis. Sobral, a city located in the northeast (equatorial zone) of Brazil (3 masculine S/40 masculine E) has 138,565 inhabitants, who are mostly white Portuguese and native Brazilian descendants. We evaluated the occurrence of hip fractures in Sobral between July 1996 and June 2000. This was a retrospective cohort-based study. Data were obtained from the medical records of the Santa Casa de Sobral, which is the reference hospital in this area. All chart records of patients aged >20 years with a diagnosis of femoral or hip fracture were revised. A total of 79 fractures were identified. Ten cases (12.6%) that occurred in patients <50 years of age were excluded from further analysis. Sixty-nine cases (87.3%) occurred in patients aged >50 years, comprising 19 (27.5%) men and 50 (72.4%) women. This gives an age-adjusted annual incidence rate of 5.59/10,000 per year in men and 12.4/10,000 per year in women, respectively, for people >50 years of age. Two patients, aged 79 and 82, died 9 and 6 days, respectively, after hospital admission. The average time of hospitalization was 7.5 days (range 4-19 days). As expected, there was a significant rise in hip fractures in people >50 years old. Interestingly, the hip fracture rate in this population is considerably lower (around 4 times less) than the reported rates in white Caucasians. PMID- 12730761 TI - Impact of subsidizing effective anti-osteoporosis drugs on compliance with management guidelines in patients following low-impact fractures. AB - Early in 2000, proven-effective antiresorptive drugs (alendronate and raloxifene) were included in the national "health basket" in Israel. We carried out the present study to evaluate the effect of subsidizing antiosteoporosis drugs on the use of antiosteoporosis drugs in patients following low-impact fractures. The rates of dispensation of antiosteoporosis drugs, in the hospital and in the community, before and after an incident of a newly diagnosed low-impact fracture, respectively, were evaluated during January and February 1998 and 1999 ("pre basket") and the corresponding months of 2000 and 2001 ("post-basket"). The study was carried out in a 950-bed teaching hospital, the only one serving the area, and the largest health maintenance organization in the area. Hospital charts of women and men age 50 years and older with new fractures following low- or moderate-impact trauma treated in the emergency room, or admitted to the orthopedic surgery and rehabilitation departments, were reviewed. A centralized pharmacy computerized database was used to follow antiosteoporosis drug dispensation in the community. A significant, approximately two-fold, increase in the baseline (before fracture) rate of osteoporosis drug dispensation was observed between the pre- and post-basket periods. The rate of patients treated after a fracture incident also increased significantly, 1.6 fold, in the post basket period; however, even in the post-basket period, two-thirds of the patients remained untreated following a fracture incident, and most of those treated received only calcium and vitamin D; only 17% received potent antiosteoporosis drugs. In a multivariate analysis, female gender, hospitalization, having the incident of fracture in the post-basket period, and above all being treated for osteoporosis before the fracture incident, had the greatest effect on the likelihood of being treated following a low-impact fracture incident. The increase in the pooled use of antiosteoporosis drugs and/or calcium/vitamin D supplements was continuous, and subsidizing created no step-up effect, besides a transient increase in the use of potent antiosteoporosis drugs in the first year following the health-basket amendment. We conclude that while subsidizing may have a significant, positive effect on antiosteoporosis drug utilization, other factors may be even more important. There is an ongoing need to find ways to encourage the use of effective pharmacological interventions for primary and secondary prevention of osteoporotic fractures. PMID- 12730763 TI - Comparison of calcium and alfacalcidol supplement in the prevention of osteopenia after kidney transplantation. AB - The aim of this observational study was to compare the effect of calcium and alfacalcidol supplementation on the regression of hyperparathyroidism and on prevention of osteopenia in patients up to 3 years after renal transplantation. Two historical cohorts were compared for that purpose. One hundred and fifty-nine patients received calcium carbonate supplement (group 1), while 81 patients were treated with alfacalcidol (group 2). Serum Ca, phosphate (P), Mg, creatinine, alkaline phosphatase (AP) and parathyroid hormone (PTH) levels were determined before and after transplantation in the two groups, for 3 years. Femoral neck and lumbar spine bone mineral density (BMD) was measured only at 3 and 6 months and 1, 2 and 3 years after transplantation. At baseline there was no difference in age or sex ratio, but prevalence in post-menopausal women was higher in group 1 (6.9% versus 1.2%). Duration on dialysis was comparable but prevalence of interstitial and undetermined nephropathies was higher in group 1. Baseline serum concentrations of PTH, Ca and P were comparable in both groups. After transplantation, plasma creatinine decreased to comparable levels in both groups. Immunosuppression by triple therapy was more prevalent in group 2, so that cumulative dose of steroid was higher in group 1, especially at 1 month because of higher incidence of acute rejections (51% versus 13%). Mean intact PTH levels decreased in both groups, from 18 pmol/l to 8.4 and 7.9 at 3 years, but the decrease was significantly greater with alfacalcidol at 6 and 12 months. At 3 months, BMD were comparable at both sites. From 3 months to 3 years after kidney transplantation, mean lumbar spine BMD significantly increased from 0.963 to 1.054 g/cm(2) in group 1, whereas there was no significant decrease (1.048 to 1.006 g/cm(2)) in group 2, the difference in changes being significant ( P<0.05). Femoral neck BMD was not significantly increased in either group (0.932 to 0.993 g/cm(2) in group 1, and 0.850 to 0.907 g/cm(2) in group 2). Expressed as percentages, these changes were +9.4% and -4% for lumbar BMD and +6.5% and +6.7% for femoral neck, for groups 1 and 2, respectively. Prevalence of osteopenia was not significantly lower at 3 years in group 1 (45% and 51%) than in group 2. During the follow-up period, osteonecrosis was diagnosed in six patients (3.8%) in group 1 and in nine (11%) in group 2. In conclusion, alfacalcidol compared to CaCO3 supplement suppressed hyperparathyroidism more rapidly and strongly. In spite of higher osteopenia risk in the CaCO3 group, lumbar BMD increase was greater and incidence of osteonecrosis higher in this group, suggesting better bone protection with CaCO3 than with alfacalcidol. PMID- 12730762 TI - Effects of dietary nutrients and food groups on bone loss from the proximal femur in men and women in the 7th and 8th decades of age. AB - We measured the impact of diet, anthropometry, physical activity and lifestyle variables on rates of hip bone mineral density (BMD) loss in 470 white men and 474 white women aged 67-79 years at recruitment dwelling in the community. The subjects were recruited from a prospective population-based diet and cancer study (EPIC-Norfolk) in Eastern England. Dietary intake was measured at baseline using 7-day food diaries and used to calculate intakes of some 31 nutrients and 22 food groups. Standardised questionnaires were used to collect data on anthropometry, physical activity and lifestyle variables. BMD loss (percent per annum; % p.a.) was measured using dual-energy X-ray absorptiometry performed on two occasions an average of 3 years apart (range 2-5 years). The mean rate of BMD change at the total hip region was -0.17% p.a. (SD 1.3% p.a.) in men and -0.41% p.a. (SD 1.2% p.a.) in women. In both men and women, weight gain protected against (and weight loss promoted) BMD loss ( P<0.0001). Markers of current physical activity were protective. In men, an increase of 1 l/s in FEV(1) was associated with an increase in BMD at an average rate of 0.25% p.a. ( P=0.013). In women, for every ten trips made per day climbing a flight of stairs, BMD increased at a rate of 0.22% p.a. ( P=0.005) and additionally a 10% increase in activities of daily living score was associated with BMD increasing at a rate of 0.12% p.a. ( P=0.011) in women. Nutritional variation appeared to have less impact on BMD loss. In men there was no evidence of an effect of any of the nutrients evaluated. However, in women, low intake of vitamin C was associated with faster rate of BMD loss. Women in the lowest tertile (7-57 mg/day) of vitamin C intake lost BMD at an average rate of -0.65% p.a., which was significantly faster compared to loss rates in the middle (58-98 mg/day) and upper (99-363 mg/day) tertiles of intake, which were -0.31% p.a. and -0.30% p.a., respectively ( P=0.016). There was no effect of fruits and vegetables, combined or separately, on rate of BMD loss. The results confirm that weight maintenance (or gain) and commonly practiced forms of physical activity appear to protect against BMD loss in this age group. Measures such as ensuring good general nutrition to guard against weight loss in the non-overweight elderly and maintenance of physical fitness could be valuable in protecting against BMD loss. The protective effect of vitamin C in women needs to be further investigated in other prospective cohort or intervention studies. PMID- 12730764 TI - Abnormalities of the PTH-vitamin D axis and bone turnover markers in children, adolescents and adults with cystic fibrosis: comparison with healthy controls. AB - Abnormalities of calcium and vitamin D metabolism in cystic fibrosis (CF) are well documented. We tested the hypothesis that alterations in calcium metabolism are related to vitamin D deficiency, and that bone resorption is increased relative to accretion in patients with CF. Calcitropic hormones, electrolytes, osteocalcin (OC) and bone alkaline phosphatase (BAP), (markers of bone mineralisation), urinary deoxypyridinoline [total (t) Dpd, a marker of bone resorption] and lumbar spine bone mineral density (LS BMD), expressed as a z score, were measured in 149 (81 M) CF and 141 (61 M) control children aged 5.3 10.99 years, adolescents aged 11-17.99 years and adults aged 18-55.9 years. Data were analysed by multiple regression to adjust for age. In patients, FEV(1)% predicted and CRP (as disease severity markers), genotype and pancreatic status (PS) were recorded. The distribution of PTH differed between groups ( P<0.0001), with CF levels both below and above the control range. 25OH vitamin D (25OHD) was not different in control and CF subjects ( P=0.06). Active hormonal vitamin D (1,25(OH)(2)D) was lower in the CF group ( P<0.0001), not explained by 25OHD or disease severity, as was serum magnesium ( P<0.0001). OC was decreased in CF adults ( P=0.004), and tDpd increased in CF adolescents ( P=0.003) and adults ( P=0.03). The ratio of OC to tDpd (a measure of bone coupling) was similar in CF and control children, but decreased in CF adolescents ( P=0.04) and adults ( P=0.02), suggesting decreased overall bone accrual in CF adolescents and uncoupling of bone balance in adults. 1,25(OH)2D was weakly correlated with OC in CF children ( r=0.43, P=0.01), and with tDpd in CF and control adolescents ( r=0.33, P=0.05 and r=0.36, P=0.02, respectively); thus there was limited evidence of association of calcitropic hormones, which had an abnormal pattern in all age groups, with bone turnover. There was no association between calcitropic hormones or bone turnover markers and LS BMD z-score. Despite vitamin D sufficiency, abnormalities of calcium metabolism and bone turnover markers were still apparent and bone accretion was decreased relative to resorption in the CF adolescent and adult groups. These changes were not fully explained by disease severity or genotype, but are consistent with reports of decreased BMD and unique bone histomorphometry in older subjects with CF. PMID- 12730765 TI - Bone mineral density and fractures among alcohol-dependent women in treatment and in recovery. AB - Women are at higher risk for osteoporosis, but most of the literature examining the effect of alcohol abuse on bone mineral density (BMD) has been in men. The aim of this study was to determine differences in BMD and fracture prevalence among women in treatment for alcohol abuse, in recovery and non-alcohol-dependent women. This cross-sectional study was completed at two residential substance abuse centers in Iowa (USA). The patients were Caucasian women, aged 18-70 years, in treatment for alcohol abuse and dependence ( n=228); in recovery and abstaining from alcohol ( n=156); and women with no history of alcohol abuse ( n=447). The main outcome measures were femoral neck and lumbar spine BMD measured by dual-energy X-ray absorptiometry (DXA); self-reported lifetime fracture prevalence. After adjusting for age and menopausal status, women in treatment had BMDs that were 7.7% ( p<0.01) and 6.3% ( p<0.01) lower at the femoral neck and lumbar spine, respectively, than non-alcohol-abusing women, and 4.8% lower at both bone sites ( p<0.01) than women in recovery. Femoral neck BMD of women in recovery was 3.1% lower ( p<0.01) than in non-alcohol-dependent women; however, the difference was not significant following multivariate analysis. Women in treatment and recovery reported more fractures during childhood and early adolescence than non-alcohol-dependent women ( p<0.01). Women in recovery also reported significantly greater numbers of fractures following sobriety than their paired non-alcohol-dependent counterparts. Alcohol abuse and dependence was associated with lower femoral neck and lumbar spine BMD. Women with histories of alcohol dependence had a higher lifetime prevalence of fractures, including time periods before the onset of problem drinking and following abstinence, suggesting that factors other than acute intoxication contributed to the greater fracture prevalence. PMID- 12730766 TI - A technical and clinical evaluation of digital X-ray radiogrammetry. AB - Skeletal assessment by morphometry at peripheral sites (e.g. metacarpal index), although simple to perform and widely available, was limited by poor precision and technical aspects of radiogrammetry. Digital X-ray radiogrammetry (DXR) uses the principles of this long-established method but applies more sophisticated methodology to overcome these problems. The aims of this study were to (a) investigate the effects of radiographic technique on bone mineral density (BMD) measurement by DXR, (b) compare DXR to dual-energy X-ray absorptiometry (DXA) and single-energy X-ray absorptiometry (SXA) and (c) determine the applicability of DXR in identifying individuals who most appropriately might be referred for axial DXA. Different radiographers performing the radiograph do not adversely affect precision. Precision, unstandardised (CV%) and standardised (sCV%), is good with both double (DF)- and single (SF)-sided emulsion radiographic film, but better with SF (CV% 0.92 vs 1.12 DF; SCV% 1.76 vs 2.93 DF). Repeat analysis precision was determined on SF (CV% 0.24, sCV% 0.55). A significant ( p<0.001), systematic difference was found between BMD measured from DF and SF (mean difference 0.017 g/cm(2)). The overall percentage difference between the methods was 2.98% (range 0.18-5.78%). Correlations between DXR BMD and DXA were moderately good (r=0.56 0.77, p<0.001); with SXA of the forearm they were excellent (r=0.91, p<0.001). The sensitivity and specificity of DXR for detecting women with osteopaenia or osteoporosis (DXA T-score less than -1; World Health Organisation) was determined at the spine [area under curve (AUC)=0.82, standard error (SE)=0.04], femoral neck (AUC=0.84, SE=0.04) and total hip (AUC=0.84, SE=0.04). Based on femoral neck BMD for detection of osteopaenia, a DXR T-score threshold of -1.05 would be appropriate for detection of patients who might benefit most from axial DXA measurements. The DXR is quick and simple to use, having potential for application in a variety of settings as analysis can be performed in a central unit, with radiographs taken in sites over a wide geographical area. Retrospective analysis may also be performed, e.g. on radiographs taken to monitor rheumatoid arthritis. The technique may also provide a simple, widely available and relatively inexpensive method to assess patients at risk of osteopaenia or osteoporosis, and who most appropriately could be referred for axial DXA. This may be particularly relevant in those who suffer low-trauma fractures and attend accident and emergency or fracture clinics, where investigation for osteoporosis is often overlooked. PMID- 12730767 TI - Risk factors for Colles' fracture in men and women: results from the European Prospective Osteoporosis Study. AB - The aim of this study was to determine the contribution of constitutional and lifestyle variables on the subsequent risk of distal forearm (Colles') fracture in a multinational, multicenter, population-prospective study. A total of 15,745 subjects from the European Vertebral Osteoporosis Study, who had completed a baseline questionnaire on lifestyle and constitutional factors, were followed up annually using a validated questionnaire to ascertain the occurrence of new fractures. Risks are expressed as hazard ratios (with 95% confidence intervals) derived from a Cox proportional hazards regression model. The incidence of Colles' fracture was 1.7 and 7.3 per 1000 person years in men and women, respectively. In women delayed menarche, over the age of 15 years, was associated with a modest increased risk [HR 1.5 (range 1.1-2.0)]. Regular walking in that group also increased the risk [HR 1.6 (1.2-2.2)] perhaps reflecting the increased exposure to risk of falling. None of the other factors examined revealed any important influences. The results are broadly in line with the few other published prospective studies suggesting only a modest role for these factors in influencing susceptibility to fracture. PMID- 12730769 TI - Design and methodology of the phase 3 trials for the clinical development of strontium ranelate in the treatment of women with postmenopausal osteoporosis. AB - The phase 3 program for strontium ranelate, a new oral agent in the treatment of women with postmenopausal osteoporosis, was aimed to assess the efficacy and safety of the daily oral dose of 2 g. This program was conducted in 12 countries, involved 75 centers, and was structured in 3 studies: FIRST (Fracture International Run-in for Strontium ranelate Trial), SOTI (Spinal Osteoporosis Therapeutic Intervention study) and TROPOS (TReatment Of Peripheral OSteoporosis). FIRST, a run-in open study, was designed to start the normalization of the calcium and vitamin D status of the patients, check all entry criteria, and ensure inclusion of a sufficient number of well-motivated patients in either one of the two therapeutic intervention protocols, SOTI or TROPOS: FIRST included 9,196 patients. SOTI and TROPOS were prospective, randomized, double-blind clinical trials comparing, in two parallel groups, the daily oral dose of 2 g of strontium ranelate with placebo, the patients of both groups receiving calcium and vitamin D according to their own deficiencies. The main objective of SOTI and TROPOS was to demonstrate a reduction in the incidence of postmenopausal women experiencing a new osteoporotic fracture (vertebral fracture in SOTI and nonvertebral fracture in TROPOS) over a 3-year treatment period, the total duration of the studies being 5 years. SOTI included 1,649 women with at least one osteoporotic vertebral fracture at inclusion and a lumbar BMD or =30 kg/m2) postmenopausal women who were either receiving hormone replacement therapy [HRT(+)] or not [HRT(-)] (total of six groups). Body weight, BMI, total body bone mineral content (TBBMC), and markers of bone formation (serum osteocalcin) and bone resorption (urinary pyridinoline (PYD) and deoxypyridinoline) were retrospectively analyzed in 210 postmenopausal women. The mean age was 67+/-6 years, with mean body weight of 70.8+/-14.2 kg, ranging from 45.0 to 115.5 kg. Body weight was positively correlated with TBBMC ( r=0.50, p<0.0001). There was a lower TBBMC and higher bone formation rate in normal weight than obese HRT(-) women, but in women taking HRT there were no differences between BMI categories. In addition, in normal-weight HRT(-) women only, PYD and body weight showed a negative correlation (r=-0.39, p=0.01). Among normal-weight, but not overweight or obese subjects, we observed higher TBBMC and lower bone turnover in the HRT(+) compared with the HRT(-) group. Regression models explained 36% of the variance in TBBMC, mainly through body weight. Additional models could only explain 11-15% of the variance in bone turnover. Taken together, these data suggest that among normal-weight but not obese postmenopausal women, higher bone turnover is associated with lower bone mass, and that only normal-weight women show a different bone turnover profile with HRT treatment. Body weight should be considered an important factor in bone metabolism with relevant clinical implications. PMID- 12730782 TI - Patient assessment using standardized bone mineral density values and a national reference database: implementing uniform thresholds for the reimbursement of osteoporosis treatments in Belgium. AB - Dual-energy X-ray absorptiometry (DXA) devices from the three main manufacturers provide different bone mineral density (BMD) values, due in part to technical differences in the algorithms for bone mineral content (BMC) and area measurements and in part to the use of different manufacturer-derived reference databases. As a result, significant differences exist between Hologic, Lunar and Norland systems in the reported young normal standard deviation scores or T scores. In a number of European countries, including Belgium, a T-score below 2.5 is one of the key criteria for reimbursement of osteoporosis treatments. This paper addresses the first attempt to implement a nationwide, uniform expression of BMD in patients, in order to harmonize drug reimbursement. To this end, measures were taken to implement a uniform expression of BMD in Belgian patients, by converting each manufacturer's absolute BMD to standardized BMD (sBMD) values and by establishing a single national reference range. PMID- 12730783 TI - Bone turnover markers during lactation, postpartum amenorrhea and resumption of menses. AB - Changes in bone turnover, and consequent bone loss and recovery during lactation and the postweaning period, are likely modulated by varying estrogen levels inherent in these time periods. To address this question we measured serum biochemical markers of bone formation (bone-specific alkaline phosphatase, amino terminal propeptide of type I procollagen, osteocalcin), of bone resorption (type I collagen carboxy-terminal telopeptide), and serum female sex hormones (estradiol, luteinizing hormone and follicle-stimulating hormone) in 32 healthy mothers prospectively after delivery, 3 months postpartum, after postpartum amenorrhea and 1 year after resumption of menses. During postpartum amenorrhea (mean 5.7, SD 2.9 months) bone mineral density decreased significantly, some 2% at the lumbar spine and some 3% at the femoral neck, but subsequently recovered completely at the former site and partially at the latter. Bone turnover marker levels were elevated at parturition and still at the end of postpartum amenorrhea. Subsequent to parturition the bone resorption marker level showed a decreasing trend while the formation marker levels continued increasing, and eventually coincided with the resorption level within the very first months postpartum. Both lactation and hormonal status modulated bone turnover marker levels. Maternal age was positively associated with increased bone turnover. Interestingly, higher parity and longer history of previous lactation were associated with lower bone turnover marker levels postpartum as compared with previously nulliparous women of the same age. The regression models explained typically some 20-30% of the variability in the bone turnover marker levels. The dynamic pattern in bone turnover is dissimilar to that occurring at menopause and it indicates that the bone loss most likely occurs in the beginning of postpartum period. It also seems that estrogen has a specific influence on bone turnover only during the first months of lactation. PMID- 12730784 TI - Is the calcium receptor a molecular target for the actions of strontium on bone? AB - The extracellular calcium-sensing receptor (CaR) plays key roles in maintaining extracellular calcium homeostasis by enabling several of the cells and tissues involved in this process to sense small changes in Ca(2+)(o) and to respond with changes in cellular function that will restore Ca(2+)(o) to its normal level. The chief cells of the parathyroid gland and the thyroidal C-cells, for example, respond to decreases in Ca(2+)(o) with increased secretion of the Ca(2+)(o) elevating hormone, parathyroid hormone (PTH), and decreased secretion of the Ca(2+)(o)-lowering hormone, calcitonin, respectively. The cells of the renal distal tubule are likewise capable of sensing Ca(2+)(o) and respond to decreases in Ca(2+)(o) with increased tubular reabsorption of Ca(2+) and vice versa, alterations in tubular function that will contribute to normalization of Ca(2+)(o). The skeleton also plays key roles in maintaining Ca(2+)(o) homeostasis and both osteoblasts and osteoclasts can sense Ca(2+)(o), with elevations in Ca(2+)(o) promoting bone formation and inhibiting bone resorption. It has been suggested that Sr(2+) could act on bone via the CaR; however, the molecular mechanisms through which Ca(2+)(o) and Sr(2+)(o) exert these actions on bone cells remain controversial. Therefore, identifying their molecular target(s) would have significant implications for the treatment of bone loss. Ideally, therapies should simultaneously inhibit bone resorption while stimulating bone formation. Administration of strontium produces exactly those effects. Previous studies with dispersed bovine parathyroid cells as well as a preliminary study using CaR-transfected Chinese hamster ovary (CHO) cells indicate that Sr(2+)(o) is an agonist of the CaR, albeit with slightly lower efficacies and potencies than Ca(2+)(o). Given that Sr(2+)(o) is distributed preferentially in bone, therefore, an action of this divalent cation on the CaR in bone cells represents one possible mechanism by which strontium ranelate, a new antiosteoporotic drug, exerts it skeletal actions in vivo. PMID- 12730785 TI - Effect of previous and present physical activity on bone mass in elderly women. AB - The importance of a physically active lifestyle as a preventive measure for maintaining bone mass is often stressed. The aim of this study was to evaluate the effect of everyday physical activity on bone mass from young adulthood to old age in 75-year-old women. Bone mineral density (BMD) was assessed by dual-energy X-ray absorptiometry technique in total body, hip (femoral neck and trochanter), and spine (LII-LIV) in a population-based cohort of 995 women, all 75 years old. Each woman responded to a questionnaire giving an estimate of the previous and present physical activity level at work and leisure time from youth and up to the present age of 75 years. No correlations were found between the different estimates of physical activity level and BMD. In a multiple regression analysis the different estimates of physical activity level did not have any combined effect on BMD in any of the skeletal regions measured; neither did the physical activity, previous or present, differ when the subgroup of women with a BMD in the highest quintile at all sites was compared with women with a BMD in the lowest quintile. The activity level in the various areas of life did not correlate to each other indicating that few women had chosen a generally active vs non-active lifestyle. A hypothetical positive effect on bone by one activity could therefore be diminished by another. The answer alternatives were mostly not normally distributed, which could also affect the results, making it difficult to identify women with extremely high or low physical activity level. In conclusion, the results from this study of randomly selected elderly women cannot confirm an effect of previous and present everyday physical activity on bone mass. Neither was it possible to find an effect of physical activity in those women with the highest BMD at all sites, suggesting that other factors are of greater importance for maintaining bone mass in elderly women. PMID- 12730787 TI - Total dose incurred by patients and staff from BMD measurement using a new 2D digital bone densitometer. AB - Dual energy X-ray absorptiometry (DXA) is a widely used and precise technique for non-invasive assessment of bone mineral density. The DXA systems have evolved from pencil X-ray beam (single detector) to fan beam (linear array detector) and recently cone beam densitometers (bi-dimensional detector), allowing for an examination to occur without any scanning and with a short acquisition time. The purpose of this study was to evaluate patient and staff dose from a new cone beam densitometer, the DMS Lexxos. Measurements were performed on a DMS Lexxos bone densitometer prototype. An anthropomorphic phantom and thermoluminescent dosimeters were used to evaluate the effective dose. Ionization chambers and electronic personal dosimeters were used to evaluate the staff dose. The effective dose is 8.4 micro Sv for an anteroposterior spine examination and 4.8 micro Sv for a femoral neck in standard mode. The averaged scattered dose rate (ambient dose equivalent) at 1 m from the beam is evaluated at 226 micro Sv/h. Assuming six patients per hour with two views per patient, the time averaged dose rate is evaluated at 2.9 micro Sv/h. By the personal dosimeter, the staff dose (Hp 10) at 1 m from the beam is evaluated at 0.23 micro Sv per examination. For one examination, patient and staff dose from this new technology remains low: in the same range as the fan-beam densitometer. PMID- 12730786 TI - Quantitative ultrasound of the calcaneus and dual X-ray absorptiometry of the lumbar spine in assessment and follow-up of skeletal status in patients after kidney transplantation. AB - Bone loss after kidney transplantation is a significant complication of immunosuppressive treatment leading to a high prevalence of bone fracture in these patients. The purpose of this study was to determine the usefulness of quantitative ultrasound (QUS) of the calcaneus in comparison with dual X-ray absorptiometry (DXA) of the lumbar spine in determining bone status and mineral changes in patients in the first 6 months after transplantation. Forty-six patients participated in the study (25 men and 21 women; age range 26-62 years, 102+/-66 months previously on dialysis). They were treated with cyclosporine, methylprednisolone, mycophenolate mofetil, and basiliximab. The 6-month cumulative steroid dose was 24.9+/-3.7 mg/kg body weight. Calcaneal QUS (Sahara, Hologic, Waltham, Mass.) and DXA (Hologic QDR 4500) of the lumbar spine were done in all patients within 3 weeks after transplantation and 6 months thereafter. Bone mineral density (BMD) of the lumbar spine measured by DXA decreased from 0.892+/-0.137 to 0.837+/-0.126 g/cm2 (p<0.0001) and the T score decreased from 1.84+/-1.29 standard deviation (SD) to 2.35+/-1.19 SD (p<0.0001) in the first 6 months after transplantation. The QUS parameters of the calcaneus were broadband ultrasound attenuation (BUA), speed of sound (SOS), and quantitative ultrasound index (QUI). The QUS parameters did not change significantly after the first 6 months. All QUS parameters correlated significantly with DXA BMD of the lumbar spine immediately after transplantation and 6 months thereafter. Significant decrease of the lumbar spine BMD in the first 6 months after transplantation was not accompanied by significant changes of calcaneal QUS parameters. The calcaneal QUS does not reflect bone mineral changes occurring in the lumbar spine and could not be a substitute for a direct-site DXA of the lumbar spine in the early period after kidney transplantation. PMID- 12730789 TI - First-degree relatives of persons suffering from osteoporosis: beliefs, knowledge, and health-related behavior. AB - OBJECTIVE: The aim of the present study was to compare the beliefs, knowledge, and health-related behaviors of first-degree relatives (FDRs) of osteoporotic women with a comparison group of women not having a FDR with osteoporosis. RESEARCH DESIGN: Telephone interviews. PARTICIPANTS: Participants were 172 women (48 FDRs and 124 comparisons), recruited by a systematic sampling from the list of all women aged 40+ attending an outpatient screening program at a large tertiary medical center in Israel. METHODS: Information was collected regarding participants' perceived susceptibility to develop osteoporosis, perceived concerns, and perceived prevention optimism. Additionally, information was collected regarding knowledge about osteoporosis, and participation in screening and preventive behavior. RESULTS: Participants in the FDR group reported higher perceived susceptibility and higher concerns about developing the disease than participants in the comparison group. A higher proportion of FDRs reported engaging in screening behavior, but no differences were found in preventive behaviors. Screening behavior was associated with perceived concerns and age for members of the comparison group, and with age for FDRs. CONCLUSIONS: FDRs are characterized by greater perceived vulnerability and concerns. However, they are no more likely to engage in preventive behaviors or have more knowledge about the disease. These findings stress the importance of expanding efforts to increase the knowledge and awareness of at-risk persons. PMID- 12730788 TI - Device-specific weighted T-score for two quantitative ultrasounds: operational propositions for the management of osteoporosis for 65 years and older women in Switzerland. AB - The World Health Organization (WHO) criteria for the diagnosis of osteoporosis are mainly applicable for dual X-ray absorptiometry (DXA) measurements at the spine and hip levels. There is a growing demand for cheaper devices, free of ionizing radiation such as promising quantitative ultrasound (QUS). In common with many other countries, QUS measurements are increasingly used in Switzerland without adequate clinical guidelines. The T-score approach developed for DXA cannot be applied to QUS, although well-conducted prospective studies have shown that ultrasound could be a valuable predictor of fracture risk. As a consequence, an expert committee named the Swiss Quality Assurance Project (SQAP, for which the main mission is the establishment of quality assurance procedures for DXA and QUS in Switzerland) was mandated by the Swiss Association Against Osteoporosis (ASCO) in 2000 to propose operational clinical recommendations for the use of QUS in the management of osteoporosis for two QUS devices sold in Switzerland. Device specific weighted "T-score" based on the risk of osteoporotic hip fractures as well as on the prediction of DXA osteoporosis at the hip, according to the WHO definition of osteoporosis, were calculated for the Achilles (Lunar, General Electric, Madison, Wis.) and Sahara (Hologic, Waltham, Mass.) ultrasound devices. Several studies (totaling a few thousand subjects) were used to calculate age adjusted odd ratios (OR) and area under the receiver operating curve (AUC) for the prediction of osteoporotic fracture (taking into account a weighting score depending on the design of the study involved in the calculation). The ORs were 2.4 (1.9-3.2) and AUC 0.72 (0.66-0.77), respectively, for the Achilles, and 2.3 (1.7-3.1) and 0.75 (0.68-0.82), respectively, for the Sahara device. To translate risk estimates into thresholds for clinical application, 90% sensitivity was used to define low fracture and low osteoporosis risk, and a specificity of 80% was used to define subjects as being at high risk of fracture or having osteoporosis at the hip. From the combination of the fracture model with the hip DXA osteoporotic model, we found a T-score threshold of -1.2 and -2.5 for the stiffness (Achilles) determining, respectively, the low- and high-risk subjects. Similarly, we found a T-score at -1.0 and -2.2 for the QUI index (Sahara). Then a screening strategy combining QUS, DXA, and clinical factors for the identification of women needing treatment was proposed. The application of this approach will help to minimize the inappropriate use of QUS from which the whole field currently suffers. PMID- 12730790 TI - Epidemiology of hip and wrist fractures in Cameroon, Africa. AB - Osteoporosis and fragility fractures are believed to be uncommon in Africa. To reevaluate this notion, we documented all patients aged 35 years and older admitted to the two main urban hospitals in Cameroon following a diagnosis of fracture during 2 years. Among 513 patients sustaining fractures (192 women, 321 men), 13.5% of all fractures in women occurred at the hip (n=26), 4.7% at the forearm (n=9), and 81.8% (n=157) at other sites (mainly tibia and femoral shaft). In men, the corresponding figures were 9% (n=29), 1.9% (n=6), and 89.1% (n=286). Of the hip and wrist fractures occurring in women, 80.0% were low energy trauma fractures due to falls, 8.6% were high-energy trauma fractures (road accidents), and 11.4% were undefined. In men, the corresponding figures were 42.9%, 34.3%, and 22.9%. Using the 1997 estimates of the population, the annual incidence rates of low-energy trauma fractures (per 100,000 persons over 35 years and above) were 4.1 in women, 2.2 in men for hip fractures, 1.2 in women, and 0.2 in men for wrist fractures. The pattern of most of the hip and wrist fractures in women is consistent with underlying bone fragility. The low incidence of fragility fractures is confirmed and is likely to be, in part, the result of reduced longevity as only 1.1% of women and 0.7% of men survive beyond 65 years of age. PMID- 12730791 TI - Skeletal status in males aged 7-80 years assessed by quantitative ultrasound at the hand phalanges. AB - Skeletal status in males has been assessed by quantitative ultrasound in only a few studies. There is the lack of data concerning age accretion, peak values, and age-decrease of bone parameters in a wide age range. The aim of this study was evaluation of a large, male, normal population (n=1,175) aged 7-80 years, by the use of quantitative ultrasound (QUS), to estimate peak value of phalangeal ultrasound parameter, age-related changes during growing and aging, the influence of body size, and determination of male normative data. In the study, amplitude dependent speed of sound (Ad-SoS) at the hand phalanges was measured using DBM Sonic 1200 (IGEA, Italy). The precision assessed by root-mean-square coefficient of variation (RMS-CV) was 0.7%. Ad-SoS ranged from 1,916 m/s in the youngest subgroup (7 years of age) to 1,910 m/s in the oldest (75-80 years of age), and the peak value (2,135 m/s) was achieved in 28-year-old males. Ad-SoS did not increase significantly to the age of 13. Ad-SoS increased significantly between subgroups aged 13 and 14 years ( p<0.0001), and at 15 and 16 years of age ( p<0.000001). A significant decrease was noted between the following age subgroups: 35-39 years vs 40-44 years ( p<0.05), 45-49 years vs 50-54 years ( p<0.000001), and 60-64 years vs 65-69 years ( p<0.001). A difference in Ad-SoS values between the youngest subgroup and peak value was 219 m/s, and an increase per year was about 10.4 m/s. From the peak value to the value in the oldest group Ad-SoS dropped by 225 m/s, and the decrease per year was about 4.6 m/s. Multiple stepwise regression analyses of Ad-SoS with age, weight,and height were performed separately for age ranges: 7-13 years (before an increase in Ad-SoS, n=299), 14 28 years (from the onset of the fast increase in Ad-SoS to the peak value, n=370) and for subjects older than 28 years (after an achievement of peak value of Ad SoS, n=506). The following equations of the multiple stepwise regression were obtained, respectively: Ad-SoS (m/s)=1,624-1.0 x age (years) -2.94 x weight (kg) + 3.06 x height (cm), r=0.48, p<0.000001, SEE=32.0; Ad-SoS (m/s)=1,533+6.87 x age (years) + 2.41 x height (cm) 0.07 x weight (kg), r=0.56, p<0.000001, SEE=54.3; Ad SoS (m/s)=1,895-3.87 x age (years) - 1.43 x weight (kg) + 2.53 x height (cm), r=0.73, p<0.00001, SEE=54.3. In conclusion, the study results allowed us to determine normative data, to assess a peak Ad-SoS value, its age-related changes and the influence of body size in the male population. PMID- 12730792 TI - Cost-effectiveness of hip protectors in institutional dwelling elderly. AB - OBJECTIVE: To assess the effectiveness and cost-effectiveness of hip protectors in the prevention of hip fracture in an elderly population living in institutions. DESIGN AND SETTING: Meta-analysis followed by economic analysis. METHODS: The meta-analysis of randomized controlled clinical trials was based on the recommendations of the Cochrane Collaboration. Model-based economic analysis was conducted using estimates of transition probabilities, costs, and utilities relevant to Canada. RESULTS: The results indicate that hip protectors compared with control results in a relative risk of hip fracture of 0.40 (95% Confidence Interval 0.23-0.70). The economic analysis found that the use of hip protectors is expected to be both effective and cost saving. Results were robust to a range of analysis exploring the uncertainty of input parameters. CONCLUSIONS: There is sufficient clinical and economic evidence to support the use of hip protectors for institutional dwelling elderly. PMID- 12730793 TI - Differential effects of hormone replacement therapy on bone mineral density and axial transmission ultrasound measurements in cortical bone. AB - The menopause has a large effect on bone density, and hormone replacement therapy (HRT) has been shown to be an effective treatment for preventing postmenopausal bone loss. The aim of this study was to compare the effects of HRT use on speed of sound (SOS) measurements at the radius, tibia, phalanx, and metatarsal with bone mineral density (BMD) measurements of the lumbar spine and proximal femur. The study population consisted of 278 healthy premenopausal women, 194 healthy postmenopausal women, and 126 healthy postmenopausal women currently receiving HRT for one or more years. SOS measurements were taken at the radius, tibia, phalanx, and metatarsal using the Sunlight Omnisense, and BMD measurements at the lumbar spine and proximal femur using Hologic QDR-4500 densitometers. Z-scores were calculated using the postmenopausal control group. Z-score differences between the postmenopausal controls and HRT group, for the entire group and with the HRT group subdivided into three groups based on duration of HRT usage, were calculated. Significant postmenopausal bone loss was found for all SOS and BMD measurements. A positive effect of HRT usage was found for all SOS measurement sites and lumbar spine BMD, although only the radius and tibia SOS and lumbar spine BMD reached statistical significance. The Z-score differences between the two groups were 0.44, 0.37, 0.15, and 0.26 for the radius, tibia, phalanx, and metatarsal SOS respectively, and 0.28, 0.00, and -0.03 for the lumbar spine, femoral neck, and total hip BMD respectively. A clear effect of the duration of HRT use was seen for the radius measurements, the differences being less marked elsewhere. In conclusion, these results demonstrate a positive effect of HRT on SOS measurements at the radius and tibia and BMD measurements of the lumbar spine. PMID- 12730794 TI - The effect of age, weight, and lifestyle factors on calcaneal quantitative ultrasound: the ESOPO study. AB - Quantitative ultrasound (QUS) techniques have been shown to be as good as bone mineral density (BMD) assessed by dual-energy X-ray absorptiometry (DXA), in predicting fracture risk: QUS technique could increase substantially the accessibility to a reliable bone osteoporosis risk evaluation, but little is know regarding the relationship of QUS to risk factors that have been found to predict DXA-BMD values and this is even more true in men. We studied 6,811 postmenopausal women 40 to 80 years of age and 4,981 men 60-80 years of age representative of the general population of all regions of Italy. All participants were questioned on lifestyle habits and on their medical history. After a physical examination "bone stiffness" (called here for simplicity, stiffness), which is derived from the values of broadband ultrasound attenuation (BUA) and speed of sounds (SoS) was measured by a heel QUS device (Achilles apparatus, Lunar, Madison, USA). The most common recognized determinants of bone mass (either categorical or continuous variables) were modeled with stiffness by multiple regression analysis or ANOVA. Stiffness was strongly related to age and weight. After adjusting for these variables, the women who had taken hormone replacement therapy for more than a year had significantly higher stiffness values. The difference versus nonusers remained significant for up to 20 years-since-menopause (YSM). This effect was so strong that for further analysis these women were excluded. By multivariate analysis, stiffness was then found to be significantly related to recalled body weight at 25 years of age, actual and past cigarettes smoked per day, and dairy calcium intake. Stiffness was also associated with a number of categorial factors adjusted for age, weight, and YSM: prior ovariectomy, history of more than 2 months confined to bed, outdoor physical activity, smoking, chronic use of any drug, and past corticosteroid use. All these categorial and continuous variables predicted stiffness equally in men and women. In conclusion, QUS bone measurements discriminate postmenopausal women according to past use of hormone replacement therapy. Risk factors usually associated to BMD as measured by DXA are also associated to calcaneal bone stiffness as measured by QUS, and most risk factors for osteoporosis usually observed in women are equally applicable to men. PMID- 12730795 TI - Increased cancellous bone in the femoral neck of patients with coxarthrosis (hip osteoarthritis): a positive remodeling imbalance favoring bone formation. AB - Osteoporosis is caused by an imbalance between bone resorption and formation which results in an absolute reduction in bone mass. In a previous study we highlighted a condition, osteoarthritis of the hip (coxarthrosis, cOA), where an imbalance between resorption and formation provided beneficial effects in the form of an absolute increase in bone mass. We demonstrated that the femoral neck in patients with cOA had increased cancellous bone area, connectivity and trabecular thickness which might contribute to the protection against fracture associated with the condition. The aim of the present study was to analyze forming and resorbing surfaces in coxarthritic cancellous bone to assess whether increased formation or reduced resorption could be responsible for these structural changes. Whole cross-sectional femoral neck biopsies were obtained from 11 patients with cOA and histomorphometric parameters compared with 14 age- and sex-matched cadaveric controls. The ratio of osteoid surface to bone surface was 121% ( p<0.001) higher in the cases but there was no significant difference in resorptive surface. The percentage osteoid volume to bone volume (%OV/BV; +270%, p<0.001) and osteoid width (O.Wi; +127%, p<0.001) were also higher in the cases. This study suggests that the increased cancellous bone mass seen in cases of cOA is due to increased bone formation rather than decreased bone resorption. Investigation of the cellular and biochemical basis for these changes might provide new insights into the pathogenesis of osteoarthritis and highlight novel biological mechanisms regulating bone multicellular unit (BMU) balance that could be relevant to developing new interventions against hip and other osteoporotic fractures. PMID- 12730796 TI - Cigarette smoke extract inhibits chemotaxis and collagen gel contraction mediated by human bone marrow osteoprogenitor cells and osteoblast-like cells. AB - Cell migration and matrix remodeling are key events in tissue repair and restructuring. Osteoblasts are responsible for the production of new bone matrix during bone remodeling. The activity of these cells can be modulated by a number of factors. The current study evaluated the hypothesis that cigarette smoke extract can alter repair and remodeling responses of human osteoprogenitor cells and osteoblast-like cells and, therefore, could explain one mechanism by which cigarette smoking leads to osteoporosis. Human osteoprogenitor cells were isolated from normal human bone marrow and maintained in culture under either control conditions or conditions that induced differentiation into osteoblast like cells. Both cell types migrated toward fibronectin and PDGF-BB as chemoattractants. Neither responded to TGF-beta1. The osteoprogenitor cells were more active in their chemotactic response. The chemotactic response of both cell types was inhibited by cigarette smoke extract in a concentration-dependent manner. Both cell types, when cultured in three-dimensional native collagen gels maintained in floating culture, induced contraction of their surrounding matrices. Contraction was augmented by serum, PDGF-BB, and TGF-beta1. Osteoprogenitor cells were less active in inducing contraction than were osteoblast-like cells. Contraction of both cell types was inhibited by cigarette smoke extract. Cigarette smoke extract also inhibited the production of fibronectin by both cell types maintained in three-dimensional culture. Addition of exogenous fibronectin partially restored the ability of the cells to contract three-dimensional collagen gels. The current study demonstrates that cigarette smoke can interfere with the ability of bone cells to participate in repair and remodeling events. Such an effect may be one mechanism leading to the development of osteoporosis. PMID- 12730797 TI - Strontium ranelate phase 2 dose-ranging studies: PREVOS and STRATOS studies. AB - The aim of the PREVOS study (PREVention Of early postmenopausal bone loss by Strontium ranelate) and the STRATOS study (STRontium Administration for Treatment of OSteoporosis) was to determine the minimum dose at which strontium ranelate (SR) is effective in, respectively, the prevention of bone loss in early postmenopausal nonosteoporotic women and the treatment of postmenopausal vertebral osteoporosis. Both studies were randomized, double-blind, placebo controlled, dose-finding studies in parallel groups and lasted 2 years. In the PREVOS study, 160 early postmenopausal women were randomized to receive placebo, SR 125 mg/day, 500 mg/day or 1 g/day. In the STRATOS study, 353 osteoporotic postmenopausal women with at least one previous vertebral fracture and a lumbar T score <-2.4 were randomized to receive placebo, SR 500 mg/day, 1 g/day or 2 g/day. In both studies, the primary efficacy parameter was lumbar bone mineral density (BMD) measured by dual-energy X-ray absorptiometry. Secondary efficacy criteria included incidence of new vertebral deformities (in the STRATOS study only) and biochemical markers of bone metabolism. In the PREVOS study, the increase in lumbar BMD from baseline in the 1 g/day group (+5.53%) was significantly different from the decrease in the placebo group ( p<0.001). In the STRATOS study, the annual increase in lumbar BMD in the 2 g/day group (+7.3% per year) was significantly higher than in the placebo group ( p<0.001). There was a significant reduction in the number of patients experiencing new vertebral deformities in the second year of treatment in the 2 g/day group (relative risk: 0.56; 95% confidence interval: 0.35, 0.89). In both studies, there was a significant increase in the bone formation marker (bone alkaline phosphatase) in the higher-dose group. Urinary excretion of the marker of bone resorption (cross linked N-telopeptide) was lower with SR than with placebo in the STRATOS study. SR was very well tolerated in both studies. The minimum dose at which SR is effective in preventing bone loss in early postmenopausal nonosteoporotic women and in the treatment of postmenopausal osteoporosis is 1 g/day and 2 g/day, respectively. PMID- 12730799 TI - The mineralization of bone tissue: a forgotten dimension in osteoporosis research. AB - Osteoporosis treatment should not only prevent the loss of bone tissue, not interfere with apatite and avoid bone mineral changes at the crystal level, but should also increase the mechanical resistance of bone and thus protect the skeleton against new fractures. Mineral substance is crystallized as nonstoichiometric carbonated apatite ionic crystals of small size and extended specific surface. Consequently, they have a very large interface with extracellular fluids, and numerous interactions between ions from the extracellular fluid and ions constituting apatite crystals are thus possible. It is generally agreed that bone strength depends on the bone matrix volume and the microarchitectural distribution of this volume, while the degree of mineralization of bone tissue is almost never mentioned as a determinant of bone strength. We now have evidence that the degree of mineralization of bone tissue strongly influences not only the mechanical resistance of bones but also the bone mineral density. In adult bone, our model is based on the impact of changes in the bone remodeling rate on the degree of mineralization of bone tissue. The purpose of this paper is to report the main results concerning the interactions of strontium (Sr) with bone mineral in animals and in osteoporotic women treated with strontium ranelate (SR). These studies aimed to evaluate using X-ray microanalysis, X-ray diffraction and computerized quantitative contact microradiography: (1) the relative calcium and Sr bone content, (2) the distribution of Sr in compact and cancellous bone, (3) the dose dependence of the deposition of Sr in bone, (4) the interactions between Sr and mineral at the crystal level (in monkeys), (5) the influence of Sr on the mean degree of mineralization of bone tissue and on the distribution of the degree of mineralization of bone tissue, and (6) the bone clearance of Sr over short periods of time (6 and 10 weeks) after cessation of SR administration (monkeys treated for 13 and 52 weeks, respectively). In monkeys killed at the end of exposure (13 or 52 weeks), Sr was taken up in a dose-dependent manner into compact and cancellous bone, with a higher content in new bone than in old bone. The Sr content greatly decreased (about 2-fold) in animals killed 6 or 10 weeks after the end of treatment but this affected new bone almost exclusively. After SR treatment, there were no significant changes in crystal characteristics. Easily exchangeable in bone mineral, Sr was slightly linked to crystals by ionic substitution (generally 1 calcium ion substituted by 1 Sr ion in each unit cell). The degree of bone mineralization was not significantly different in the various groups of monkeys. Thus, at the end of long-term SR treatment and after a period of withdrawal, Sr was taken up in a dose-dependent manner into new bone without alteration of the degree of bone mineralization and with no major modification of bone mineral at the crystal level. In postmenopausal osteoporotic women treated with SR (0.5, 1 and 2 g/day) for 2 years, Sr was dose-dependently deposited into new bone without changes in the degree of mineralization of bone tissue. These findings could reflect dose-dependent stimulation of bone formation and are of potential value for the use of SR in the treatment of osteoporosis. In conclusion, the different studies performed on bone samples from monkeys and humans treated with various doses of SR showed that Sr was heterogeneously distributed between new and old bone but in a dose-dependent manner without alteration of the crystal characteristics and the degree of mineralization of bone tissue, even after long-term administration of often high doses of SR (the highest therapeutic dose used in humans is 4-fold lower than the lowest experimental dose administered to monkeys). This emphasizes the value, as antiosteoporotic treatment, of SR, which is safe at the bone mineral level. PMID- 12730800 TI - Bone strength and its determinants. AB - Osteoporosis is a disease defined by decreased bone mass and alteration of microarchitecture which results in increased bone fragility and increased risk of fracture. The major complication of osteoporosis, i.e., fracture, is due to a lower bone strength. Thus, any treatment of osteoporosis implies an improvement in bone strength. Bone strength is determined by bone geometry, cortical thickness and porosity, trabecular bone morphology, and intrinsic properties of bony tissue. Bone strength is indirectly estimated by bone mineral density (BMD) using dual-energy X-ray absorptiometry (DXA). Since DXA-measured BMD accounts for 60-70% of the variation in bone strength, some important factors are not captured by DXA in the progression of osteoporosis and the effects of antiosteoporotic treatment. Geometry and trabecular microarchitecture have also to be taken into account. Thus, the assessment of intrinsic mechanical quality of bony tissue should provide a better understanding of the role of tissue quality in determining bone strength. The careful investigation of all the determinants of bone strength (bone tissue included) should be considered in the pathophysiology of osteoporosis and in the mechanisms of action of antiosteoporotic drugs. PMID- 12730798 TI - Assessment of prevalent and incident vertebral fractures in osteoporosis research. AB - Vertebral fractures are the hallmark of osteoporosis, and it has been shown that the presence of vertebral fractures adds to the risk of future osteoporotic fractures. Thus, the evaluation of spinal radiographs for prevalent or incident vertebral fractures is important in the clinical evaluation of patients with osteoporosis, in clinical drug trials for osteoporosis treatment and in the epidemiologic evaluation of populations at risk for osteoporosis. Traditionally, conventional lateral radiographs of the thoracolumbar spine have been analyzed visually by radiologists or experienced clinicians to identify vertebral fractures in patients with clinical indications. Epidemiologic studies or clinical drug trials in osteoporosis research have considerably different requirements and expectations. To reduce potential subjectivity in qualitative readings and to provide definable, reproducible and objective methods to detect vertebral fracture, and to accommodate the assessment of large numbers of radiographs, various morphometric approaches have been explored and employed. However, potential deficiencies in morphometry have led to a renewed interest in standardized qualitative visual approaches for defining vertebral fractures and visual criteria. Numerous studies have shown that semiquantitative interpretation, after careful centralized training and standardization, can produce results with excellent intra- and interobserver reproducibility. We firmly believe that the experience from several studies has shown that there is a major role for radiologists and clinicians alike to carefully assess and diagnose vertebral fractures using standardized grading schemes such as the one proposed in this review. In the context of epidemiologic studies and clinical drug trials, quantitative morphometry may be used; however, the studies would be flawed if quantitative morphometry were to be performed in isolation without additional adjudication by a trained and highly experienced radiologist or clinician. PMID- 12730801 TI - Optimizing bone metabolism in osteoporosis: insight into the pharmacologic profile of strontium ranelate. AB - Strontium ranelate (SR) is currently being developed for the treatment of osteoporosis. Pharmacologic studies in animal models have shown that its efficacy on bone mass is based on its original mode of action on bone formation and bone resorption. In normal mice, SR increased bone formation and vertebral bone mass. In normal rats, SR increased bone mass and the mechanical properties of vertebral, humeral and femoral bones, associated with increased femoral shaft diameter. Vertebral bone mineral density and bone strength were also increased by SR, whereas stiffness was not altered, underlining that the improvement in bone strength occurs without inducing defective bone mineralization. In normal adult monkey alveolar bone, SR decreased bone resorption and increased bone. In ovariectomized (OVX) rats, SR limited the reduction in bone mineral content and the decrease in trabecular bone volume induced by estrogen deficiency, by inhibiting bone resorption while maintaining bone formation. Curative treatment with SR also partially restored bone mass in OVX rats. In the model of hind limb immobilization in rats, SR reduced bone resorption and partially limited long bone loss, as assessed by bone mineral content, bone volume, and histomorphometric and biochemical indices of bone resorption. The unique mode of action of SR on bone formation and resorption is also supported by in vitro studies. In calvaria culture systems and osteoblastic cell cultures, SR enhanced the replication of preosteoblastic cells and consequently increased collagen synthesis. Moreover, SR inhibited the bone-resorbing activity of isolated mouse osteoclasts and devreased osteoclast differentiation markers in chicken bone marrow cultures. Altogether, these pharmacologic results suggest that SR optimizes bone metabolism by decreasing bone resorption and promoting bone formation, which may be of potential value in the treatment of osteoporosis. PMID- 12730802 TI - Computerized heart allograft-recipient monitoring: a multicenter study. AB - Computerized heart allograft recipient monitoring (CHARM) is a unique concept of patient surveillance after heart transplantation (HTx), based on the evaluation of intramyocardial electrograms (IEGMs) recorded non-invasively with telemetric pacemakers. Previous open, single-center studies had indicated a high correlation between CHARM results and clinical findings. The present study was initiated to assess the suitability of CHARM for monitoring the absence of rejection in a blind, multicenter context. During the HTx procedure, telemetric pacemakers and two epimyocardial leads were implanted in 44 patients at four European HTx centers. IEGMs during pacing were recorded and transferred via the Internet to the CHARM computer center, for automatic data processing and extraction of diagnostically relevant information, i.e., the maximum slew rate of the descending part of the repolarization phase of the ventricular evoked response (VER T-slew). The study period comprised the first 6 months after HTx, during which the transplant centers were blind to the CHARM results. A single threshold diagnosis model was prospectively defined to assess the ability of the VER T-slew to indicate clinically significant rejection, which was defined as an endomyocardial biopsy (EMB) grade greater than or equal to 2, according to the grading system of the International Society for Heart and Lung Transplantation. All EMB slides from three centers were reviewed blind by the pathologist of the fourth center in order that agreement among the histological diagnoses at the various centers could be assessed. Totals of 839 follow-ups and 366 EMBs were obtained in 44 patients. Thirty-seven patients were alive at the end of the study period. Age at HTx, EMB grade distribution, and rejection prevalence varied significantly between the centers. Review of the EMB results showed considerable differences with respect to classification of significant rejection. Comparison of average VER T-slew values with and without rejection in the 15 patients who exhibited both states revealed significantly lower values under the influence of rejection (97+/-13% vs 79+/-15%, P<0.0001). Twenty out of the 25 cases with significant rejection were correctly identified by VER T-slew values below a threshold of 98% (sensitivity =80%, specificity =50%, negative predictive value =97%, positive predictive value =11%; P<0.0005). Of the EMBs, 48% could have been saved if the diagnosis model had been used to indicate the need for EMB. A high negative predictive value for the detection of cases with significant rejection has been obtained in a prospective, blind, multicenter study. The presented method can, therefore, be used to supplement patient monitoring after HTx non invasively, in particular to indicate the need for EMBs. In centers with patient management similar to the ones who participated in the study, this may allow a reduction in the number of surveillance EMBs. PMID- 12730803 TI - Regulation of pro-inflammatory and anti-inflammatory cytokine responses by Kupffer cells in endotoxin-enhanced reperfusion injury after total hepatic ischemia. AB - The effects of Kupffer cells on cytokine responses in endotoxin-enhanced reperfusion injury after total hepatic ischemia were investigated in this study. Male rats pretreated with either normal saline solution (NS group) or gadolinium chloride (GdCl(3)) to inhibit Kupffer cell function (GC group) were subjected to 60 min of hepatic ischemia. These animals received either normal saline solution or sublethal doses of endotoxin (1 mg/kg) at reperfusion. In the NS group, endotoxin administration induced an enhanced tumor necrosis factor-alpha (TNF alpha) and interleukin-10 production 1 h after reperfusion with a subsequent peak of macrophage inflammatory protein-2 (MIP-2) levels, which resulted in a 7-day survival rate of 30%. Despite endotoxin administration, GdCl(3) pretreatment significantly suppressed TNF-alpha and increased interleukin-10 production 1 h after reperfusion, which led to a decline in MIP-2 production and amelioration of functional and structural liver damage with a 7-day survival rate of 80%. Augmented pro-inflammatory and anti-inflammatory cytokine responses by Kupffer cells were associated with endotoxin-enhanced reperfusion injury after hepatic ischemia. Kupffer cell blockade has a potential to attenuate the insult via modulation of cytokine responses. PMID- 12730804 TI - The impairment of hepatocytes and sinusoidal endothelial cells during cold preservation in rat fatty liver induced by alcohol and the beneficial effect of hepatocyte growth factor. AB - A fatty liver resulting from alcohol intake is often unattractive for grafting. In this study, we investigated the impairment of hepatocytes and sinusoidal endothelial cells (SECs) during cold preservation of alcohol-induced fatty liver and examined the efficacy of human recombinant hepatocyte growth factor (hrHGF). Rats were fed an alcohol diet. We performed histological examinations of the hepatocytes and observed the ultrastructural alteration of the SECs. Additionally, we measured hepatic transaminase and peroxidative lipids for hepatocellular injury and the hyaluronic acid uptake rate (HUR) to determine SEC injury. We added hrHGF to University of Wisconsin (UW) solution to assess the protective effect of the agent. Numerous fatty deposits were observed in ethanol induced fatty livers. These grew with the duration of cold storage. Hepatic transaminases of the effluents increased during cold preservation in the livers of alcohol-treated rats. Additionally, peroxidative lipids in the effluents increased during cold preservation in the livers of alcohol-treated rats, whereas they were undetectable in non-alcohol-treated rat livers. The sinusoidal endothelium had severely deteriorated in the livers of alcohol-treated rats. Further, the HUR decreased with ethanol treatment and/or cold preservation. The addition of hrHGF suppressed the increase of hepatic transaminase in the effluent of cold-preserved alcohol-treated livers. Peroxidative lipids in the same effluents were undetectable. In fatty livers, both hepatocytes and SECs received severe damage during cold preservation. Furthermore, we demonstrated that hepatocellular injury was significantly inhibited by hrHGF. PMID- 12730805 TI - Cardiovascular 24-h rhythms and renal excretory function in rats after allogeneic kidney transplantation. AB - We investigated long-term and circadian blood-pressure changes in a Fischer- >Lewis rat model of chronic renal allograft rejection. Ten days after allogeneic kidney transplantation, rats were equipped with telemetry transmitters for continuous monitoring of blood pressure. Urine was sampled 24, 96 and 180 days after kidney transplantation for measurement of renal protein loss and excretion of nitric oxide metabolites. Proteinuria increased with time after transplantation, while urinary excretion of nitric oxide metabolites declined. In five of six rats blood pressure remained in the normotensive range, and its 24-h pattern was preserved up to 6 months post-transplantation. One animal showed hypertensive blood pressure and a disturbed 24-h pattern with peak values during the rest period. In the post-mortem analysis this rat showed a hydronephrotic kidney graft. In conclusion, blood pressure and its 24-h pattern are preserved after successful allogeneic kidney transplantation in the rat. PMID- 12730806 TI - Hepatic-artery aneurysm in adult liver transplantation. AB - Hepatic artery aneurysm (HAA) is a rare vascular complication, but has a high mortality rate in liver transplant recipients. This study reports the precipitating factors, clinical manifestation, pre-operative diagnosis, related micro-organism, management, and outcome, in a series of HAAs that developed after adult orthotopic liver transplantation (OLT). Data on the primary disease as well as on the above were obtained from a prospective database, and all case records were reviewed. There were eight (0.5%) HAAs in 1,575 adult cadaveric OLTs between 1982 and March 2001. All were pseudo-aneurysms around the native hepatic-artery (HA) anastomosis, and all occurred in whole-organ OLTs. There were three types of clinical presentations: sudden hypotension (n=4), gastrointestinal (GI) bleeding (n=2), and abnormal liver-function tests (LFTs) (n=2). The majority (n=7) presented within the first 2 months (median: 27.5 days, range: 12-760 days) following OLT. A pre-operative diagnosis of HAA was not determined in five cases. The sensitivity of abdominal ultrasound scan (USS), computed tomography (CT) scan and angiography for detection of HAAs was 3 of 5, 1 of 2 and 3 of 4, respectively. Micro-organisms could be identified in six patients (bacteria n=4 and fungi n=3). All patients underwent urgent operations (excision of HAA in six and ligation in two cases). Immediate reconstruction of the HA was carried out, two different methods being used: repair of native arteries (n=2) and arterial conduit (interposition n=3 and aortic conduit n=2). Two patients died peri operatively, two died within 2 months, and the remaining four patients are alive at between 8.6 and 12.8 years after repair. HAA following OLT is unpredictable in its presentation, and the sensitivity of clinical and radiological detection is low. A high index of suspicion is required, and urgent surgery with immediate re vascularisation and use of appropriate antibiotic/anti-fungal agents is recommended. PMID- 12730807 TI - Modulation of matrix gelatinases and metalloproteinase-activating process in acute kidney rejection. AB - Changes in matrix metalloproteinase (MMP) activities would contribute to the accumulation of extracellular matrix during acute kidney allograft rejection. MMP 2 and MMP-9 and other gelatinolytic activities were examined in the rejected graft and the urine of a rat model of acute kidney rejection (orthotopic allotransplantation from a Buffalo donor to a Wistar-Furth recipient) by either zymography or fluorescence assay. MMP-2, membrane type 1 (MT1)-MMP, and tissue inhibitor of metalloproteinase (TIMP)-2 were also examined by immunodetection. The proMMP-2 activity and protein level increased in the graft during rejection when compared with normal Buffalo kidney, whereas activated MMP-2 decreased. TIMP 2 protein levels were markedly decreased and MT1-MMP proteolytic fragments (44-40 kDa) were undetectable. This suggests an altered MT1-MMP-dependent processing of proMMP-2 into active MMP-2 due to a diminished TIMP-2 level in acute kidney rejection. In the urine the overall gelatinolytic activity decreased considerably, although activity associated with an as yet unidentified 78-kDa protein appeared 6 days after transplantation. PMID- 12730809 TI - Living-donor liver transplantation for homozygous familial hypercholesterolemia from a donor with heterozygous hypercholesterolemia. AB - Familial hypercholesterolemia is a rare inherited disease with an incidence of approximately one per million. Severe hypercholesterolemia is observed from the time of birth onwards. It is associated with severe atherosclerosis in childhood, leading to death from myocardial infarction before the age of 20 years. Liver transplantation is the only effective treatment for this disease. We experienced the case of an infant aged 2 years 5 months who had homozygous familial hypercholesterolemia and who received a liver graft from his father, who had familial heterozygous hypercholesterolemia. The pre-operative plasma cholesterol level was >800 mg/dl. After liver transplantation, the recipient's cholesterol level decreased to 250 mg/dl after we administered the HMG-CoA reductase inhibitor. At present, 6 months after transplantation, the patient is doing well and free from a special diet. We can thus conclude that the combination therapy of liver transplantation from a donor with heterozygous familial hypercholesterolemia on cholesterol-lowering drugs is an effective therapy for a patient with the homozygous type of hypercholesterolemia. PMID- 12730808 TI - Applicability of liver transplantation in Catalonia at the end of the millennium. A prospective study of adult patient selection for liver transplantation. AB - We prospectively studied the global applicability of liver transplantation in Catalonia, a region with a high rate of organ donation. We followed 232 adult patients assessed as possible candidates for liver transplantation over 12 months in the three hospitals that perform the procedure in this region. The liver disease leading to patient assessment was cirrhosis in most cases, alone (159 patients) or associated with hepatocellular carcinoma (57 patients). After being assessed, 150 patients (65%) were accepted for transplantation and included on the waiting list, and 82 (32%) were excluded. Death during the period of assessment, advanced tumoral disease, early stage of liver disease, and extrahepatic co-morbidities were the most important reasons for exclusion. The median time of assessment of patients accepted for transplantation was 40 days. Of the 150 patients included on the waiting list, 131 (87%) received transplants, 17 (11%) were removed from the list, and two were still waiting for transplantation at the end of the follow-up period. Death and tumor progression were the most important reasons for patients' removal from the waiting list. The median time on the waiting list was 59 days. In conclusion, among liver transplant candidates the overall applicability of this therapy in Catalonia was relatively low (131 out of 232 transplant candidates finally underwent transplantation, 56%), and inadequate liver-transplant indications and death or tumor progression during the period of assessment or while the patient was on the waiting list were the most frequent reasons why liver transplantations did not proceed. PMID- 12730810 TI - Invasive aspergillosis in solid-organ transplantation: report of eight cases and review of the literature. AB - Invasive fungal infections are life-threatening complications in solid-organ transplantation. Although the rate of fungal infections in transplant recipients is lower than that of other infections, the mortality rate is higher. The most frequent fungi isolated from these kinds of infections are Candida spp. and Aspergillus spp. We retrospectively evaluated the clinical and laboratory findings in eight patients who were treated for invasive aspergillosis (IA) at our center. This report describes these cases and discusses the relevant literature. PMID- 12730811 TI - The effects of intracoronary administration of vitamin E on myocardial ischemia reperfusion injury during coronary artery surgery. AB - BACKGROUND: Vitamin E has a strong antioxidant capacity, and has been used in several ischemia-reperfusion studies. The aim of this study was to investigate the effects of water-soluble vitamin E (alpha-tocopherol) on myocardial protection during coronary artery surgery. METHODS: Water soluble vitamin E (100 mg) in tepid saline (n = 14) or tepid saline alone (n = 16) was administered into the coronary arteries at the end of aortic cross-clamping. Cardiac troponin-I (cTn-I), MB-isoenzyme of creatine kinase (CK-MB), myoglobin, blood gas, and lactate levels in systemic and coronary sinus blood and hemodynamic variables were assessed during and after the operation. RESULTS: Eight hours after reperfusion, cTn-I levels increased to 3.06 +/- 1.8 ng/ml and 6.97 +/- 3.9 ng/ml in the vitamin E group and control group, respectively (p = 0.01). Coronary sinus lactate concentration was 2.68 +/- 0.5 mmol/l in the vitamin E group and 4.01 +/- 1.5 mmol/l in the control group 60 minutes after reperfusion (p = 0.027). CONCLUSIONS: Administration of vitamin E into the coronary arteries before removal of the aortic cross-clamp can reduce myocardial cell injury and protect the myocardium from ischemia-reperfusion injury. PMID- 12730813 TI - Limb trauma with arterial injury: long-term performance of venous interposition grafts. AB - BACKGROUND: In contrast to arterial occlusive disease, data on long-term outcomes after vein grafts in limb trauma with arterial injury are sparse. PATIENTS: From 1991 through 2001, 22 trauma victims received 23 interposition vein grafts performed by an interdisciplinary team of trauma and vascular surgeons. Indications included both blunt and penetrating injuries with critical limb ischemia in the majority of cases. RESULTS: Operative treatment of the injured vessels (brachial n = 5, radial/ulnar n = 7, popliteal n = 6, tibial n = 3, pedal n = 2) encompassed venous interposition graft of either saphenous (n = 15) or cephalic vein (n = 8). All patients survived the operative procedure. 4 graft occlusions were noted and 3 major amputations had to be performed (one despite patent graft). 13 patients (76%) were available for duplex ultrasound examination after a mean follow-up of 59 months where patent grafts could be detected in all cases. CONCLUSION: A multidisciplinary approach ensures optimal treatment strategy of arterial injury in extremity trauma. Interposition vein grafts provide durable long-term results and should be attempted even in single-vessel injuries of forearm and lower leg. PMID- 12730812 TI - External reinforcement of varicose veins with PTFE prosthesis in infrainguinal bypass surgery -- clinical results. AB - OBJECTIVE: Segmental varicose degeneration of the autogenous greater saphenous vein may limit its use in infrainguinal bypass surgery. Wrapping a PTFE prosthesis around dilated veins has emerged as an option to create externally reinforced vein bypasses. Results regarding graft patency and limb salvage were analyzed. METHODS: Between September 1995 and January 2001, 35 infrainguinal bypass operations in 33 patients were performed with greater saphenous veins exhibiting segmental varicose dilatation. Grafts were followed by duplex scan and retrospective analysis of graft patency and limb salvage was performed. RESULTS: One bypass prompted successful revision for early occlusion. Four bypasses required additional reintervention during follow-up. 48 months primary, primary assisted and secondary patency rates were 66%, 82% and 82%, respectively, with a limb salvage rate of 97%. Duplex scan failed to demonstrate stenosis of the reinforced vein segments or aneurysmal degeneration of the residual vein. CONCLUSION: External reinforcement with a PTFE prosthesis allows the use of autogenous greater saphenous veins with varicose dilatation and enables the construction of all autogenous bypasses with promising graft patency and limb salvage. PMID- 12730814 TI - EuroSCORE as a predictor for complications and outcome. AB - OBJECTIVE: The purpose of this study was to evaluate the usefulness of EuroSCORE in terms of prediction of the outcome as a result of preoperative information in a cohort of patients. METHODS: We analyzed the data from 751 patients treated between Jan 1 and Dec 31, 1998. We used contingency tables and applied methods of discriminant analysis for the evaluation. RESULTS: Compared to the 14.799 patients from whose data the EuroSCORE system had originally been derived [1,2], we had a smaller portion of patients in the low-risk group (24.5% vs. 30.6%), a comparable portion of patients in the medium-risk group (42.2% vs. 40.5%) and a higher proportion of patients in the high-risk group (33.2% vs. 29.0%). This difference in the risk distributions was highly significant (p < 0.001). The application of the EuroSCORE system showed that deaths only occurred in the high risk group in our hospital. We had 36 deaths amongst the patients, which gives an overall mortality rate of 4.9% and a mortality rate of 1.6% in the medium-risk group and of 12.4% in the high-risk group. The discriminant analysis showed that with the EuroSCORE as single predicting variable - virtually all survivors were classified correctly (giving a specificity of 99.0%). The sensitivity was rather low at only 25%; however, this means that many of the high-risk patients in our clinic survived. The total misclassification rate, however, was 12.7%. Additionally, we found that a higher score value predicts a significantly higher probability for perioperative complications and also results in significantly increased average times for the bypass and total time for surgery. CONCLUSION: The EuroSCORE is a valuable score system for the prediction of the overall outcome of patients following open heart surgery, and is easy to use. As far as perioperative complications are concerned, however, some specifications seem desirable. PMID- 12730815 TI - The relevance of large strains in functional tissue engineering of heart valves. AB - BACKGROUND: Exposing the developing tissue to flow and pressure in a bioreactor has been shown to enhance tissue formation in tissue-engineered heart valves. Animal studies showed excellent functionality in these valves in the pulmonary position. However, they lack the mechanical strength for implantation in the high pressure aortic position. Improving the in vitro conditioning protocol is an important step towards the use of these valves as aortic heart valve replacements. In this study, the relevance of large strains to improve the mechanical conditioning protocol was investigated. METHODS: Using a newly developed device, engineered heart valve tissue was exposed to increasing cyclic strain in vitro. Tissue formation and mechanical properties were analyzed and compared to unstrained controls. RESULTS: Straining resulted in more pronounced and organized tissue formation with superior mechanical properties over unstrained controls. Overall tissue properties improved with increasing strain levels. CONCLUSIONS: The results demonstrate the significance of large strains in promoting tissue formation. This study may provide a methodological basis for tissue engineering of heart valves appropriate for systemic pressure applications. PMID- 12730817 TI - Video-assisted thoracic needle aspiration cytology for malignancy of the peripheral lung. AB - BACKGROUND: The technique of video-assisted thoracic fine-needle aspiration cytology (VAT-FNA) to the lung has been described in very few publications, and its diagnostic role has not been evaluated so far. Thus current studies focus on whether the diagnostic role could be applied usefully to VAT-FNA of peripheral lung lesions. METHODS: Between January 1995 and January 2000, a total of one hundred and twenty-eight cases of VAT-FNA on lung tumors were reviewed retrospectively. The superficial lung was visualized a part of directly or indirectly by scope and a 22-gauge needle inserted for FNA. Material was expressed onto glass slides, and smears were stained by our modified quick-stain method. The cytological diagnoses based on VAT-FNA were reviewed and compared with the final histopathological diagnoses. RESULTS: The cytological diagnosis was true positive in 100 cases (93.5% sensitivity), whereas the true negative result in 20 cases was 95.2% specificity. The false-positive rate was 4.8%, and false-negative results were 6.5%. The accordance in all malignant cases between cytology and histology was 73.8%. VAT-FNA caused no difficulties in any of the cases. CONCLUSION: The application of VAT-FNA to the peripheral lung lesion is not only useful, cost-beneficial, safe and minimally invasive but also accurate. Especially, this method may play an important role in cases of suspected malignancy in peripheral cases. PMID- 12730816 TI - Antibiotic prophylaxis in elective thoracic surgery: cefuroxime versus cefepime. AB - BACKGROUND: Infection is one of the major morbidity factors after thoracic surgery. Although different prophylactic regimens have been used to prevent this complication, the ideal prophylactic agent, dose and duration of administration remain unknown. METHODS: All patients included underwent elective lung resection. 102 selected patients consecutively scheduled for major thoracic surgery were enrolled in this study and randomized into either the cefuroxime group (n = 50) or the cefepime group (n = 52). RESULTS: Twelve pathologic bacterium strains were isolated in the cefepime group, whereas only 5 pathogenic strains were isolated in the cefuroxime group; the difference was statistically significant (p = 0.04). Two empyemas (3.8 %) in the cefepime group were noted, while the cefuroxime group showed no cases of empyema (p = 0.16). Overall infection rate (pneumonia + bronchopneumonia + empyema) were 14.0 % and 26.7 % in the cefuroxime and the cefepime groups, respectively (p = 0.12). Using chest radiography, pulmonary infiltration was found to be more frequent in the cefuroxime group (p=0.002). CONCLUSION: Cefuroxime as a prophylactic agent in major thoracic surgical operations was marginally more effective than cefepime, and presented an additional cost advantage. PMID- 12730818 TI - Perioperative pneumoperitoneum after lobectomy -- bilobectomy operations for lung cancer: a prospective study. AB - BACKGROUND: The aim of this study is to identify the effectiveness of perioperative pneumoperitoneum to prevent air leak after the lobectomy bilobectomy operations for lung cancer. METHODS: A prospective study was designed on consecutive 50 patients who had lobectomy-bilobectomy operations for lung cancer and whose remnant lung had failed to fill the half of the hemithoracic cavity under 30 cm H(2)O positive pressure ventilation during the operation with totally relaxed diaphragm. The patients were divided into two groups: group 1 (25 patients) with perioperative pneumoperitoneum, group 2 (25 patients) without perioperative pneumoperitoneum. RESULTS: The statistical analysis between the two groups did not show any significant difference in terms of age, preoperative FEV1, and the type of resection. Perioperative pneumoperitoneum significantly reduced the duration of postoperative air leak (2.2 +/- 1.15 days vs. 6.04 +/- 3.27 days p < 0.0001) and total chest tube drainage time (3.84 +/- 0.98 days vs. 7.88 +/- 3.16 days p < 0.001). CONCLUSION: Perioperative pneumoperitoneum after lobectomy-bilobectomy operations for lung cancer is an effective method to decrease air leak and chest tube drainage time. PMID- 12730819 TI - Isolated coronary artery rupture after blunt chest trauma. AB - After blunt chest trauma, a patient with chronic coronary heart disease sustained an isolated rupture of the right coronary artery. All findings suggested a heart contusion complicated by a non-compromising pericardial effusion and aggravated by anticoagulation with phenprocoumon. After right-ventricular failure occurred, emergency coronary revascularization could not prevent a fatal outcome. This case emphasizes that a coronary artery lesion may be considered in those cases of thoracic trauma with preexisting coronary calcification. PMID- 12730821 TI - Repair of pulmonary artery sling by reimplantation without cardiopulmonary bypass. AB - A 20-month-old girl successfully underwent repair of pulmonary artery sling through a median sternotomy by division of the left pulmonary artery and its reimplantation into the main pulmonary artery without cardiopulmonary bypass or tracheal reconstruction. The patient is doing well on 18 months follow-up with unobstructed pulmonary blood flow and dramatic reduction of tracheal stenosis. Simple repair of pulmonary artery sling is feasible with good results in selected cases without tracheomalacia. PMID- 12730820 TI - Ex situ resection of primary cardiac tumors. AB - The prognosis of malignant heart tumors is pessimistic; 50% of patients die within 6 months. No optimal therapy has been established, and standardized therapeutic concepts have not been developed due to the low incidence of this disease. In most cases, chemotherapy and radiotherapy have not shown any survival benefit compared to surgical treatment. Obviously, radical resection of the tumor is the most important determinant for long-term survival. Here, we report on two patients in whom radical resection of heart tumors could be accomplished only after explantation of the heart. PMID- 12730822 TI - A single-stage video-assisted thoracoscopic surgery procedure for simultaneous bilateral spontaneous pneumothorax in a supine position. AB - We report on the case of a 23-year-old man with simultaneous bilateral spontaneous pneumothorax (SBSP), treated with bilateral video-assisted thoracoscopic surgery (VATS) in a supine position. SBSP is a very rare condition that can be life-threatening when therapeutic techniques fail. We performed a unique operative procedure for SBSP using one-stage bilateral VATS in a supine position. This procedure is less invasive, more effective, and safer for the treatment of SBSP. PMID- 12730824 TI - Deep sternal wound infection and risk analysis -- what do we really know? PMID- 12730823 TI - Endobronchial occlusion method of bronchopleural fistula with metallic coils and glue. AB - We describe our technique for endobronchial occlusion of bronchopleural fistula (BPF) with metallic coils and glue through a fiber-optic bronchoscope under local anesthesia. After anchoring the vascular embolization coils at the fistula bronchus, cyanoacrylate glue was sprayed. The sprayed glue obliterates gaps between the coils and stabilizes them. This procedure successfully occluded fistula bronchus except in one post-pneumonectomy case of large fistula. This method may be one of therapeutic options for BPF, especially in patients at high risk for curative surgery under general anesthesia. PMID- 12730825 TI - ICD Implantation with and without combined myocardial revascularization -- incidence of ICD therapy and late survival. PMID- 12730827 TI - A novel class of tests for the detection of mitochondrial DNA-mutation involvement in diseases. AB - We develop a novel class of tests to detect mitochondrial DNA (mtDNA)-mutation involvement in complex diseases by the study of affected pedigree members. For a pedigree, affected individuals are first considered and are then connected through their relatives. We construct a reduced pedigree from an original pedigree. Each configuration of a reduced pedigree is given a score, with high scores given to configurations that are consistent with mtDNA-mutation involvement and low scores given to configurations that are not consistent with mtDNA-mutation involvement. For many pedigrees, the weighted sum of scores of the pedigrees is calculated. The tests are formed by comparing the observed score with the expected score under the null hypothesis that only nuclear autosomal mutations are involved. We study the optimality of score functions and weights under the heterogeneity model without phenocopies. We also develop a method to estimate the contribution that mtDNA mutations make if they are involved under a heterogeneity model. Finally, we apply our methods to three data sets: Leber hereditary optic neuropathy, a disease that has been proved to be caused by mtDNA mutations; non-insulin-dependent diabetes mellitus (NIDDM); and hypertension (HTN). We find evidence of mtDNA-mutation involvement in all three diseases. The estimated fraction of patients with NIDDM due to mtDNA-mutation involvement is 22% (95% confidence interval [CI] 6%-38%). The fraction of patients with HTN potentially due to mtDNA-mutation involvement is estimated at 55% (95% CI 45% 65%). PMID- 12730828 TI - Cohen syndrome is caused by mutations in a novel gene, COH1, encoding a transmembrane protein with a presumed role in vesicle-mediated sorting and intracellular protein transport. AB - Cohen syndrome is an uncommon autosomal recessive disorder whose diagnosis is based on the clinical picture of nonprogressive psychomotor retardation and microcephaly, characteristic facial features, retinal dystrophy, and intermittent neutropenia. We have refined the critical region on chromosome 8q22 by haplotype analysis, and we report the characterization of a novel gene, COH1, that is mutated in patients with Cohen syndrome. The longest transcript (14,093 bp) is widely expressed and is transcribed from 62 exons that span a genomic region of approximately 864 kb. COH1 encodes a putative transmembrane protein of 4,022 amino acids, with a complex domain structure. Homology to the Saccharomyces cerevisiae VPS13 protein suggests a role for COH1 in vesicle-mediated sorting and transport of proteins within the cell. PMID- 12730830 TI - The search for quality in surgical practice. PMID- 12730832 TI - Long-term assessment of psychologic outcomes of orthognathic surgery. AB - PURPOSE: This long-term study of post-orthognathic surgery patients aimed at assessing perceptions of problems with physical and psychologic functioning, self concept, body image, and satisfaction with the surgical outcome based on subjective evaluations. In addition, the patient's perception of self-concept, body image, and function was compared with the perception of pretreatment patients and controls with no treatment need. PATIENTS AND METHODS: One hundred seventeen patients who underwent orthognathic surgery 10 to 14 years earlier and participated in a psychologic study 10 years earlier received questionnaires that were composed of 4 instruments designed to assess these factors. A sample of 92 persons representing a population without treatment need and 39 patients requesting treatment for a malocclusion served as control groups. RESULTS: The findings of this study support the hypothesis that improvement in appearance brought about by orthognathic surgery is associated with improvement in psychosocial adjustment. Orthognathic surgery resulted in subjective estimation of function, appearance, health, and interpersonal relationships that was higher than that among pretreatment and no-treatment control groups. The level of body image and self-esteem approximated but did not reach that of a nonpatient population. CONCLUSIONS: In view of the current psychologic and social environment, patients should be offered the appropriate treatment to correct a disfigurement if it is subjectively perceived by them as a handicap, in part to improve the psychologic outcome. PMID- 12730829 TI - The structure of linkage disequilibrium at the DBH locus strongly influences the magnitude of association between diallelic markers and plasma dopamine beta hydroxylase activity. AB - There is currently a great deal of interest in using linkage disequilibrium (LD) mapping to locate both disease and quantitative-trait loci on a genomewide scale. Recent findings suggest that much of the human genome is organized in discrete "blocks" of low haplotype diversity, but the utility of such blocks in identifying genes influencing complex traits is not yet known and must ultimately be tested empirically through use of real data. We recently identified a putative functional polymorphism (-1021C-->T) in the 5' upstream region of the DBH gene that accounted for 35%-52% of the total phenotypic variance in plasma dopamine beta-hydroxylase (DBH) activity in samples from three distinct populations. In the present study, we genotyped 11 diallelic markers at the DBH locus surrounding -1021C-->T in 386 unrelated individuals of European origin. We identified a single 10-kb block containing -1021C-->T, in which four haplotypes comprised 93% of the observed chromosomes. Only markers within the block were highly associated with phenotype (P< or =2.2 x 10(-10)), with one exception. In general, association with phenotype was strongly correlated with the degree of LD between each marker and -1021C-->T. Of four LD measures assessed, d(2) was the best predictor of this relationship. Had one attempted to map quantitative-trait loci for plasma DBH activity on a genomewide basis without prior knowledge of candidate regions and not included (by chance) markers within this haplotype block, the DBH locus might have been missed entirely. These results provide a direct example of the potential value of constructing a haplotype map of the human genome prior to embarking on large-scale association studies. PMID- 12730831 TI - Recovery after third molar surgery: clinical and health-related quality of life outcomes. AB - PURPOSE: The study goal was to assess both clinical and health-related quality of life (HRQOL) outcomes after third molar surgery. METHODS: Patients who were having 4 third molars removed were enrolled in a prospective clinical trial. Baseline data were recorded that included demographics, the patient's and surgeon's assessment of third molar conditions, and details of the surgical procedure. After surgery, clinical data were collected that detailed healing and any treatment that was rendered. Each patient was given an HRQOL instrument to complete on each postsurgery day for 14 days; the instrument was designed to assess a patient's perception of recovery in 4 main categories: pain, lifestyle, oral function, and other symptoms related to the procedure. RESULTS: Recovery data were available for 630 of 740 enrolled patients. The median age of the 630 patients was 21 years, and the median operation time was 30 minutes. Recovery for most HRQOL measures occurred within 5 days after surgery. However, recovery from pain to the criterion of "little or none" was delayed relative to other HRQOL measures. Twenty-two percent of patients were treated for delayed healing after surgery. CONCLUSIONS: Having both clinical and HRQOL data on recovery after third molar surgery could assist the surgeon when informing prospective patients about what to expect after surgery to remove third molars. PMID- 12730833 TI - Removal of miniplates in maxillofacial surgery: University Hospital Birmingham experience. AB - PURPOSE: We sought to study the incidence and causes of removal of osteosynthesis miniplates (plates) in oral and maxillofacial surgery in a single unit over a 13 month period and to identify factors associated with plate removal. PATIENTS AND METHODS: We conducted a retrospective study of 172 patients attending the Department of Oral and Maxillofacial Surgery, University Hospital Birmingham, between November 1, 1998, and November 30, 1999, in whom maxillofacial osteosynthesis plates were inserted or removed. Not all patients who had plates removed had them inserted within the same time frame. RESULTS: During a period of 13 months (November 1, 1998, through November 30, 1999), 308 plates were inserted into 153 patients. During the same period, 51 plates were removed from 28 patients, of whom 9 underwent plate insertion and subsequent removal of 25 plates within the time period of the study. Thirty-four (67%) of the plates removed from 20 patients were symptomatic. Infection was the most common cause for removal, occurring in 14 patients (50% of patients who had plates removed) and accounting for 22 plates (43% of plates removed). Symptoms relating to plates necessitating removal occurred in the first year after insertion in 15 patients (53%), accounting for 21 plates. CONCLUSIONS: Our experience with the removal of miniplates is comparable with that of previous studies. Plate-related problems leading to removal are more likely to occur within the first year after insertion. PMID- 12730834 TI - Referral patterns for the treatment of facial trauma in teaching hospitals in the United States. AB - PURPOSE: The management of facial trauma is considered an integral part of the training of several specialties, including general plastic surgery, otolaryngology, and oral and maxillofacial surgery. Referral patterns of patients who have sustained facial trauma to these various specialty services, however, vary at different institutions according to physician preferences and protocols. The purpose of this project was to examine the referral patterns of facial trauma in the United States at teaching hospitals. MATERIALS AND METHODS: A questionnaire survey of physician-chiefs of emergency or trauma services at teaching hospitals was carried out. Scenarios involving a variety of facial injury patterns were presented, and a hypothetical referral was requested. In addition, questions regarding preferences and opinions regarding the various services were included. RESULTS: Most teaching hospitals had a formal protocol for the referral of patients with facial injuries. With the exception of mandible fractures, referral patterns for patients with facial injuries were relatively even across the 3 specialties. Interestingly, only 56% of respondents would seek the same referral for themselves or relatives in the same way as they would refer a patient based on their in-house protocol. In regard to timeliness, efficiency, and perceived competency in the handling of facial trauma, oral and maxillofacial surgery had statistically significant higher scores than otolarygology and plastic surgery, which were not statistically distinguishable between each other. CONCLUSIONS: All 3 specialties appear to be involved in the management of facial trauma at teaching institutions in the United States; therefore, it seems unlikely that any one specialty will be singled out as the sole provider of these services at all institutions. PMID- 12730835 TI - Primary chronic osteomyelitis in childhood and adolescence: a retrospective analysis of 11 cases and review of the literature. AB - PURPOSE: Primary chronic osteomyelitis (PCO) of the jaw is a rare, nonsuppurative, chronic inflammation of an unknown cause. The disease is not age specific. So far, only case reports of this disease with an onset in childhood or adolescence have been described. PATIENTS AND METHODS: Review of the patient data of our department from the past 30 years revealed 11 patients with an early onset of PCO in childhood and during puberty. RESULTS: Demographic data, clinical course, radiologic and histologic examinations, and treatment modalities are described and compared with the literature with special emphasis on the somewhat confusing terminology used for this pathology. CONCLUSION: We present a group of 11 patients with an early onset of PCO of the mandible, which, to our knowledge, is the largest described series to date. The series shows the complexity of this disease in a young patient population, demanding a careful evaluation of each case before initiation of therapy. PMID- 12730836 TI - Comparison of the postsurgical stability of the Le Fort I osteotomy using 2- and 4-plate fixation. AB - PURPOSE: This study was conducted to evaluate the postoperative stability of Le Fort I osteotomies accomplished with 2-plate versus 4-plate fixation. METHODS: This is a retrospective study involving 32 patients who underwent Le Fort I 1 piece osteotomy concurrent with orthodontic therapy. All patients were treated by 1 attending surgeon during an 18-month period. Sixteen patients were treated by plate and screw fixation consisting of 4 miniplates (group I), and an additional 16 patients were treated using 2 miniplates (group II). In group I, fixation was accomplished with 2.0-mm low-profile Lorenz (Walter Lorenz Surgical Inc, Jacksonville, FL) plates and screws placed at the piriform aperture and at the maxillary buttress. Four screws were placed in each of the plates. In group II, fixation was accomplished with 2.0-mm low-profile Lorenz plates and screws placed at the piriform aperture. Again, 4 screws were placed in each of the plates. RESULTS: Serial cephalometric evaluation at arbitrary anterior nasal spine and posterior nasal spine for both groups showed that postoperative skeletal changes in the direction of the surgical movement were seen in approximately 20% of cases; these changes averaged less than 1 mm. Postoperative skeletal changes opposite to the direction of the surgical movement were seen in approximately 30% of cases; these changes also averaged less than 1 mm. No postoperative skeletal changes were seen in approximately 50% of cases. For all measured changes about arbitrary anterior nasal spine and posterior nasal spine, there was no significant difference between groups I and II. CONCLUSION: This study suggested that postoperative skeletal changes associated with the use of 2-plate fixation do not appear to differ significantly from those seen with 4-plate fixation. PMID- 12730837 TI - Clear cell odontogenic carcinoma: evaluation of reported cases. AB - PURPOSE: We present the cases of 4 patients with clear cell odontogenic carcinoma and a contrasting case of a patient with a calcifying epithelial odontogenic tumor from Massachusetts General Hospital. Differential diagnosis, distinguishing characteristics, and data from 32 cases in the literature are also reported. Because this rare tumor behaves aggressively with both regional and distant spread, a better understanding of its biologic behavior is imperative. PATIENTS AND METHODS: The 4 patients with clear cell odontogenic carcinoma were women and had an average age of 65 years (range, 40 to 77 years). Three tumors were in the posterior mandible and 1 was in the maxilla. Three patients complained of pain; bony expansion and tooth mobility were noted in isolated cases. A comparison is made with other reported cases, and factors associated with recurrence and survival are presented. RESULTS: All patients were treated with resection, and 1 patient received postoperative radiotherapy. Two patients with positive surgical margins underwent further resection. All patients have been followed an average of 2 years (range, 0.5 to 4 years) and presently have no evident disease. CONCLUSIONS: Recognition of this unusual tumor and differentiating it from other clear cell neoplasms is critical in developing an appropriate treatment plan. Management should include wide en bloc resection and long-term follow-up. PMID- 12730838 TI - Expression of bone morphogenetic proteins during mandibular distraction osteogenesis in rabbits. AB - PURPOSE: We examined the expression pattern of bone morphogenetic proteins (BMPs) during mandibular distraction osteogenesis in rabbits and also investigated the mechanism of membranous bone distraction. MATERIALS AND METHODS: Twenty-three rabbits underwent mandibular distraction (protocol; no latency period, a 1-week distraction at 0.5 mm/d, and a 2-week consolidation period). Samples were collected at 3, 5, and 7 days of distraction and at 1-week and 2-week consolidation. We prepared undecalcified fresh-frozen sections and immunohistochemically evaluated the expression of BMPs 2 through 8. RESULTS: Both endochondral ossification and intramembranous ossification were observed. The expression of BMPs 2, 4, 5, and 6 was observed continuously from the beginning of distraction. BMP-7 was expressed weakly. The expression of BMP-3 was not observed conspicuously during distraction but was strongly expressed at 1- and 2-week consolidation. CONCLUSION: The expression pattern of BMPs during membranous bone distraction was similar to that during long bone distraction, but it differed from the expression pattern of long bone distraction in that the expression of BMPs was maintained for 2 weeks after the completion of distraction. PMID- 12730840 TI - Effects of propranolol or paracetamol on lidocaine concentrations in serum and tissues. AB - PURPOSE: The goal of this study was to determine the interaction of lidocaine after paracetamol or propranolol administration. MATERIALS AND METHODS: Group A rats received a mixture of cold lidocaine and (14)C lidocaine into the masseter muscle. Group B rats received lidocaine and paracetamol 7.5 mg/kg orally, whereas group C received lidocaine intramuscularly and propranolol 1.5 mg/kg orally. Five consecutive doses were administered. The levels of these drugs were estimated in serum and the total concentration and the percent protein binding of lidocaine in tissues (liver and mandible) were determined. The rats were killed 2 hours after the last dose. RESULTS: The results suggest that lidocaine concentrations in serum were significantly increased after either paracetamol or propranolol administration. Additionally, the combined therapy of propranolol and lidocaine resulted in a significant decrease in the total concentration and the percent protein binding of lidocaine in tissues. CONCLUSIONS: The coadministration of lidocaine with paracetamol or propranolol interferes with the metabolic profile, resulting in pharmacokinetic interactions that may be significant for the determination of the correct dose of lidocaine in clinical applications. PMID- 12730841 TI - Applicants' opinions about the selection process for oral and maxillofacial surgery programs. AB - PURPOSE: This study was conducted to investigate the characteristics of and opinions about the selection process of applicants to residency programs in oral and maxillofacial surgery (OMS) in the year 2000. The current investigation paralleled a survey completed in the 1970s, with additional questions to assess contemporary curriculum and surgical training evolution. Differences from the previous (1976) survey, a profile of applicants in 2000, and characteristics of OMS residency programs that applicants view as attractive are discussed. SUBJECTS AND METHODS: Questionnaires were sent to 307 applicants to OMS residency programs registered in the dental matching program. To provide a more direct comparison of the study completed in 1977, the current questionnaire used the original survey as a model. It was divided into 6 sections: general information, information from the formal application and letters of recommendation, the interview, the selection process, the characteristics applicants were seeking in a program, and a retrospective review of programs visited. RESULTS: In this study, 118 responses were received, representing 38% of the total. Respondents were predominately men (86%) and single, and attended dental school in all regions of the United States and the world. Respondents listed geographic location (65%) and national reputation (58%) as important factors for selecting programs to which to apply. Many respondents felt that a previous dental internship (85%), national board scores (83%), and class rank (79%) would be considered important in screening applicants for interview. A program's reputation, personalities of residents and attending staff, and clinical material were ranked as important factors contributing to program selection. Fifty-three of the 79 successful applicants reported that they were matched with their first-choice institution. Sixty percent of programs were considered worthy of revisiting by the respondents. CONCLUSIONS: Respondents in 2000 had more knowledge of the selection process before interviewing and submitted 2 times the number of applications than applicants in 1977. When selecting programs to which to apply, current applicants were less influenced by geographic location and the recommendations of other dentists. Applicants were attracted to programs that they perceive to have a good reputation and abundant clinical material. Residencies that convey a friendly atmosphere, favorable interpersonal skills of enrolled residents, and appealing personalities of the faculty were more attractive to candidates. PMID- 12730839 TI - Elevated blood lead resulting from maxillofacial gunshot injuries with lead ingestion. AB - PURPOSE: The purpose of this study was to identify the contribution of ingested lead particles to elevated blood lead concentrations in victims of gunshot injury to the maxillofacial region. PATIENTS AND METHODS: As part of a larger study of the effects of retained lead bullets on blood lead, a retrospective review of study findings was completed on 5 of 8 patients who sustained injuries to the maxillofacial region. These 5 patients were recruited into the larger study within 11 days of injury and showed a penetration path for the projectile that engaged the upper aerodigestive tract. All subjects were recruited from patients presenting for care of their gunshot injuries to a large inner-city trauma center with a retained bullet resulting from a gunshot injury. An initial blood lead level was measured for all recruited patients and repeated 1 to 17 weeks later. Medical history was taken along with a screening and risk factor questionnaire to determine other potential or actual sources (occupational/recreational) of lead exposure. (109)Cd K-shell x-ray fluorescence determinations of bone lead were completed to determine past lead exposure not revealed by medical history and risk factor questionnaire. Radiographs taken of the abdomen and chest, required as a part of the patient's hospital care, were retrospectively reviewed for signs of metallic fragments along the aerodigestive tract. RESULTS: All 5 patients retained multiple lead pellets or fragments at the site of injury, sustained fractures of the facial bones, and showed increases in blood lead. Three of the 5 study subjects who sustained maxillofacial gunshot injuries involving the mouth, nose, or throat region showed metallic densities along the gastrointestinal tract indicative of ingested bullet fragments. Each patient with ingested bullet fragments showed rapid elevation of blood lead exceeding 25 microg/dL and sustained increases well beyond the time when all ingested fragments were eliminated. A 3-year follow-up on these 3 patients showed significantly sustained elevation of blood lead but less than that observed during the initial 6 months after injury. None of the 5 study subjects showed any evidence of metallic foreign bodies within the tracheobronchial regions indicative of aspiration. CONCLUSION: Ingestion of lead fragments can result from gunshot injuries to the maxillofacial region and may substantially contribute to a rapid increase in blood lead level. Prompt diagnosis and elimination of ingested lead fragments are essential steps necessary to prevent lead being absorbed from the gastrointestinal tract. Increased blood lead in victims after gunshot injuries must be fully evaluated for all potential sources, including recent environmental exposure, absorption of lead from any remaining bullets in body tissues, and the possibility of mobilization of lead from long-term body stores such as bone. PMID- 12730842 TI - Histomorphometric evaluation of short-term changes in masseter muscle after lengthening the rabbit mandible by distraction osteogenesis. AB - PURPOSE: Structural changes in muscles may affect the process during and after distraction osteogenesis (DO) of the mandible. However, the response of the masticatory muscles is still not well defined after gradual lengthening of the mandible. In this experimental study, short-term structural changes in masseter muscles of the rabbits are evaluated after mandibular DO. MATERIALS AND METHODS: Left mandibles of 10 New Zealand rabbits were lengthened by DO for 7 days in the rate of 1 mm/day. Mandibles of all animals were removed at the end of the consolidation period. Muscle biopsy samples of distracted and contralateral sides were histopathologically investigated, and histomorphometric results were statistically analyzed. RESULTS: Atrophy, hypertrophy, regeneration, and concomitant mild interstitial edema and fibrosis were found more evident in experimental side biopsy samples 30 days after distraction. Histomorphometric analysis revealed a statistically significant decrease in the mean regions of masseter muscle fibers of the distracted sides compared with the control sides (P <.05). CONCLUSION: This experimental study showed that the structure of masseter muscle is influenced during and shortly after mandibular distraction osteogenesis. Atrophic changes of the ipsilateral masseter muscles may be regarded as regenerative response that occurs during and shortly after distraction period. PMID- 12730843 TI - Expansile lesion of the mandible in a 45-year-old woman. PMID- 12730844 TI - Proliferative verrucous leukoplakia: report of a case with characteristic long term progression. PMID- 12730845 TI - Papillary oncocytic cystadenoma of palatal minor salivary gland: a case report. PMID- 12730846 TI - Giant pilomatricoma (epithelioma of Melherbe): report of a case and review of literature. PMID- 12730847 TI - Shrapnel-induced sialolith--a rare etiology for sialadenitis: case report. PMID- 12730848 TI - Sagittal split osteotomy in removal of impacted third molar. PMID- 12730849 TI - The vertical ramus osteotomy for high condylar fractures. PMID- 12730855 TI - Translational research: Bridging the widening gap between basic and clinical research. PMID- 12730850 TI - Where have all the heroes gone? PMID- 12730856 TI - Image of the month: "hide-bound" bowel sign in scleroderma. PMID- 12730857 TI - Rho kinase blockade prevents inflammation via nuclear factor kappa B inhibition: evidence in Crohn's disease and experimental colitis. AB - BACKGROUND & AIMS: Rho proteins are involved in the regulation of several cellular functions. Data from in vitro studies suggest that RhoA could be involved in the inflammatory response. We investigated the role of RhoA and its downstream effector Rho kinase in intestinal inflammation. METHODS: Activation of RhoA was assessed by pull-down assays. A specific inhibitor of Rho kinase, Y 27632, was used to examine the role of Rho kinase in inflammatory response in vivo and in vitro by molecular biology and by immunological and biochemical approaches. RESULTS: Increased activation of RhoA was found in inflamed intestinal mucosa of patients with Crohn's disease and of rats with 2,4,6 trinitrobenzene sulfonic acid-induced colitis. Oral administration of Y-27632 in rats significantly reduced the colonic inflammation. In vitro, activation of RhoA alone was sufficient to induce tumor necrosis factor production. Y-27632 inhibited production of tumor necrosis factor-alpha and interleukin-1 beta by lamina propria and peripheral blood mononuclear cells. Rho kinase inhibition prevented nuclear factor kappa B activation and I-kappa B phosphorylation and degradation. We showed that Rho kinase associates with and activates I-kappa B kinase alpha and that Y-27632 prevents I-kappa B kinase activation. CONCLUSIONS: Our study provides the first evidence that Rho kinase activates I-kappa B kinase and, thus, nuclear factor kappa B, suggesting a key role of Rho kinase in inflammatory responses and intestinal inflammation. Specific inhibition of Rho kinase may be a promising approach for the treatment of patients with Crohn's disease. PMID- 12730858 TI - Hyperleptinemia prevents increased plasma ghrelin concentration during short-term moderate caloric restriction in rats. AB - BACKGROUND & AIMS: Ghrelin is an orexigenic hormone secreted by the stomach. Increased plasma ghrelin concentration was reported during diet-induced weight loss in obese humans, suggesting that ghrelin contributes to adaptive increment in appetite associated with caloric restriction. Leptin reduces spontaneous food intake and body weight in rodents. The current study tested the hypothesis that increased plasma leptin prevents the potential increase in plasma ghrelin concentration during moderate caloric restriction in lean rats. METHODS: Six month-old male rats (body weight, 367 +/- 9 grams) were randomly assigned to one of the following treatments (8 rats each) for 1 week: (1) leptin subcutaneous infusion to induce moderate hyperleptinemia and moderate caloric restriction ( 26% of ad libitum), (2) vehicle infusion and pair feeding, and (3) vehicle infusion and ad libitum feeding. RESULTS: Leptin-treated (-19 +/- 5 grams) and pair-fed (-19 +/- 2) rats lost weight compared with ad libitum-fed rats (-3 +/- 1, P < 0.05). Compared with control (6.8 +/- 0.7 ng/mL), plasma leptin was higher in leptin-treated (18.6 +/- 0.9 ng/mL, P < 0.01) rats and lower in pair-fed rats (4.3 +/- 0.4 ng/mL, P < 0.05). Plasma ghrelin was substantially higher in calorie restricted than control rats (2505 +/- 132 pg/mL vs. 1790 +/- 134 pg/mL, P < 0.01), and leptin treatment (1625 +/- 117 pg/mL) completely prevented this change. Plasma ghrelin concentration was negatively correlated with body weight changes in calorie-restricted and control (r = -0.75, P < 0.01) but not in leptin treated rats (P > 0.8). CONCLUSIONS: Moderate hyperleptinemia prevents an increase of plasma ghrelin during moderate short-term caloric restriction. Satiety-inducing effects of leptin include suppression of gastric orexigenic signals and disruption of a potential feedback mechanism between body weight changes and plasma ghrelin in lean adult rats. PMID- 12730860 TI - Increased risk of noncardia gastric cancer associated with proinflammatory cytokine gene polymorphisms. AB - BACKGROUND & AIMS: Genetic variations in proinflammatory and anti-inflammatory cytokine genes influence individual response to carcinogenic exposures. Polymorphisms in interleukin (IL)-1 beta and its endogenous receptor antagonist are associated with risk of Helicobacter pylori-related gastric cancer. The aim of this study was to evaluate the role of proinflammatory cytokine gene polymorphisms in gastric and esophageal cancers defined by anatomic subsite. METHODS: We assessed polymorphisms of the IL-1 gene cluster and 4 other cytokine genes in a population-based case-control study of upper gastrointestinal cancers, including gastric cardia (n = 126) and noncardia adenocarcinoma (n = 188), esophageal squamous cell carcinoma (n = 53), and adenocarcinoma (n = 108), and frequency-matched controls (n = 212). ORs for the different cancers were computed from logistic regression models adjusted for potential confounding factors. RESULTS: Proinflammatory genotypes of tumor necrosis factor alpha and IL-10 were each associated with more than doubling of the risk of noncardia gastric cancer. Carriage of multiple proinflammatory polymorphisms of IL-1B(o) IL-1 receptor antagonist, tumor necrosis factor A, and IL-10 conferred greater risk, with ORs (and 95% confidence intervals) of 2.8 (1.6-5.1) for one, 5.4 (2.7-10.6) for 2, and 27.3 (7.4-99.8) for 3 or 4 high-risk genotypes. In contrast, these polymorphisms were not consistently related to the risks of esophageal or gastric cardia cancers. Polymorphisms in IL-4 and IL-6 were not associated with any of the cancers studied. CONCLUSIONS: A proinflammatory cytokine genetic profile increases the risk of noncardia gastric adenocarcinoma but not other upper gastrointestinal cancers, possibly by inducing a hypochlorhydric and atrophic response to gastric H. pylori infection. PMID- 12730861 TI - Prophylaxis of pouchitis onset with probiotic therapy: a double-blind, placebo controlled trial. AB - BACKGROUND & AIMS: We have recently documented the efficacy of a highly concentrated probiotic preparation (VSL#3) in the prevention of flare-up in patients with chronic pouchitis. The aim of this study was to compare probiotic therapy with VSL#3 versus placebo in the ability to prevent the onset of acute pouchitis during the first year after ileal pouch-anal anastomosis. METHODS: Forty consecutive patients who underwent ileal pouch-anal anastomosis for ulcerative colitis were randomized to receive either VSL#3 (1 packet containing 900 billion bacteria/day) (n = 20) or an identical placebo (n = 20) immediately after ileostomy closure for 1 year. The patients were assessed clinically, endoscopically, and histologically after 1, 3, 6, 9, and 12 months. Health related quality of life was assessed using the Inflammatory Bowel Disease Questionnaire. RESULTS: Two of the 20 patients (10%) treated with VSL#3 had an episode of acute pouchitis compared with 8 of the 20 patients (40%) treated with placebo (log-rank test, z = 2.273; P < 0.05). Treatment with VSL#3 determined a significant improvement in Inflammatory Bowel Disease Questionnaire score, whereas this was not the case with placebo. CONCLUSIONS: Treatment with VSL#3 is effective in the prevention of the onset of acute pouchitis and improves quality of life of patients with ileal pouch-anal anastomosis. PMID- 12730862 TI - Plasma citrulline: A marker of enterocyte mass in villous atrophy-associated small bowel disease. AB - BACKGROUND & AIMS: Plasma citrulline, a nonprotein amino acid produced by enterocytes, was suggested as a marker of remnant enterocyte mass in patients with short bowel. Our objective was to evaluate citrulline as a marker of severity and extent of villous atrophy in patients without intestinal resection. METHODS: Forty-two patients with celiac disease and 10 patients with non-celiac villous atrophy disease were studied by plasma postabsorptive citrulline and biological dosages, biopsies of proximal (duodenojejunal) small bowel and distal ileum (n = 25), or measurement of vitamin B(12) absorption (n = 4). Nine patients were reevaluated after following a gluten-free diet for 1 year. Controls were 51 healthy subjects and 10 severely malnourished patients with anorexia nervosa with no intestinal mucosal abnormalities. RESULTS: Plasma citrulline concentration was lower (P < 0.001) in patients with villous atrophy (24 +/- 13 micromol/L) than in healthy subjects (40 +/- 10 micromol/L) and patients with anorexia nervosa (39 +/ 9 micromol/L). Three thresholds were individualized: <10 micromol/L for patients with diffuse total villous atrophy (n = 10), 10-20 micromol/L for patients with proximal-only total villous atrophy (n = 12), and 20-30 micromol/L for patients with partial villous atrophy (n = 10). Plasma citrulline concentration was correlated to the severity and extent of villous atrophy (r = 0.81; P < 0.001) and to albuminemia (r = 0.47; P < 0.01). Receiver operating characteristic curves indicated that plasma citrulline concentration was the best biological variable to predict villous atrophy. Following a 1-year gluten-free diet, plasma citrulline concentration increased in histologically responsive (n = 6) but not in unresponsive (n = 3) patients. CONCLUSIONS: In patient villous atrophy diseases, plasma citrulline concentration may prove to be a simple and reliable marker of reduced enterocyte mass. PMID- 12730863 TI - Antro-fundic dysfunctions in functional dyspepsia. AB - BACKGROUND & AIMS: Symptoms in functional dyspepsia have been related to impaired accommodation and hypersensitivity of the proximal stomach. We hypothesized that identification of putative antral dysfunctions provides a more comprehensive pathophysiological interpretation. METHODS: In 30 patients with functional dyspepsia and 22 healthy subjects, 2 consecutive studies were performed. In study I, with the subjects in the upright position, the proximal and distal stomach were selectively distended by bags containing air and water, respectively, while perception and fundic relaxation in response to antral distention were measured. In study II, by using air-filled bags connected to a tensostat, the proximal and the distal stomach were selectively distended by positioning the subjects on the right and left lateral decubitus, respectively, while perception, compliance, and the responses to intestinal nutrient infusion were measured. RESULTS: Patients with dyspepsia showed hypersensitivity of both the proximal stomach (discomfort at 30 +/- 3 g vs. 62 +/- 2 g in controls; P < 0.05) and the antrum (discomfort at 31 +/- 2 g vs. 53 +/- 4 g in controls; P < 0.05). Fundic and antral fasting tone was normal, but reflex fundic relaxation induced either by antral distention (3 +/- 16 mL at 80 mL of distention vs. 38 +/- 10 mL in controls; P < 0.05) or by intestinal nutrients (35 +/- 7 mL vs. 107 +/- 8 mL in controls; P < 0.05) was markedly impaired. CONCLUSIONS: Antral and fundic dysfunctions interact to produce the symptoms of functional dyspepsia, and impaired control of fundic accommodation may lead to overload of a hypersensitive antrum. PMID- 12730864 TI - Pain and biomechanical responses to distention of the duodenum in patients with systemic sclerosis. AB - BACKGROUND & AIMS: Abnormalities of the small intestine have been indicated in systemic sclerosis. The aim was to use a new method to study the active-passive mechanical and sensory properties of the duodenum in these patients. METHODS: A volume-controlled ramp-distention protocol was used in the duodenum in 9 patients and 8 healthy controls. The nonpainful/painful sensations, pressure, cross sectional area, wall tension, and strain were evaluated. Using butylscopolamine for muscle relaxation, the active (contractile muscular component) and passive (other mechanical tissue components) were computed. RESULTS: The contraction amplitude was smaller and the cross-sectional area higher in the patients (P < 0.05). Both the active and passive tension as function of strain was translated to the left in the patients, indicating a stiffer wall. The maximum active tension and the corresponding strain were 62% and 69% lower in the patients (P < 0.05). An association was found between the duration of the disease and the strain (P < 0.05). The perception score was higher as function of pressure, tension, and strain (P = 0.01, P = 0.03, and P < 0.01, respectively) in the patients than in the controls, with strain as the most sensitive variable to describe the sensory response. In 5 patients who complained of regular clinical symptoms, the referred pain area to distention was enlarged. CONCLUSIONS: Systemic sclerosis resulted in increased stiffness and impaired muscle function of the duodenum. The pain evoked by a controlled strain of the gut was increased and can explain many of the symptoms reported in the clinic. PMID- 12730865 TI - Folate status, genomic DNA hypomethylation, and risk of colorectal adenoma and cancer: a case control study. AB - BACKGROUND & AIMS: Low folate intake may increase risk for colorectal cancer by inducing DNA hypomethylation. This study reports the influence of folate status, DNA methylation, and polymorphisms of methylenetetrahydrofolate reductase (MTHFR 677C-->T and 1298A-->C), methionine synthase (MS 2756A-->G), and cystathionine beta-synthase (CBS 844ins68) on risk for developing colorectal neoplasia. METHODS: Thirty-five patients with adenoma, 28 patients with cancer, and 76 controls were recruited for a case control study. Recruitment consent rate was 98%. Blood samples were obtained for determination of blood folates, vitamin B(12), homocysteine, DNA methylation, and genotypes. Tissue biopsy samples were obtained at colonoscopy for determination of DNA methylation in colonic mucosa. Folate status was assessed by constructing a score from estimates of dietary intake and serum and erythrocyte folate. RESULTS: Cancer patients had 26% lower folate status (95% confidence interval [CI]: 6% to 44%, P = 0.01) and 21% lower serum vitamin B(12) concentration (95% CI: -38% to 1%, P = 0.06) compared with controls. [(3)H] methyl incorporation into colonic DNA was 26% higher in patients with adenoma (95% CI: 8% to 56%, P = 0.009) and 30% higher in patients with cancer (95% CI: -3% to 48%, P = 0.08) compared with controls. High folate status was associated with decreased risk for cancer (P = 0.01 for trend). Colonic and leukocyte DNA hypomethylation were associated with increased risk for adenoma (P = 0.02 and P = 0.01 for trend, respectively) and a nonsignificantly increased risk for cancer (P = 0.09 and P = 0.08 for trend, respectively). CONCLUSIONS: Low folate status and DNA hypomethylation are associated with colorectal neoplasia. PMID- 12730866 TI - Platelets trigger a CD40-dependent inflammatory response in the microvasculature of inflammatory bowel disease patients. AB - BACKGROUND & AIMS: Platelets circulate in an activated state in patients with inflammatory bowel disease (IBD), but their role in the pathogenesis of IBD is unclear. The recent demonstration that activated platelets express CD40 ligand (L) provides a mechanism of interaction with CD40-positive endothelial cells, inducing them to produce proinflammatory mediators. We investigated whether platelets from patients with IBD express enhanced levels of CD40L and induce human intestinal microvascular endothelial cells (HIMEC) to up-regulate cell adhesion molecule (CAM) expression and secrete chemokines. METHODS: CD40L expression was assessed in resting and thrombin-activated platelets by flow cytometry and in mucosal microthrombi by confocal microscopy. Platelet-HIMEC cocultures were used to study CAM up-regulation, and interleukin (IL)-8 and RANTES production by HIMEC. RESULTS: IBD platelets expressed significantly higher CD40L levels than those of healthy subjects, and CD40L-positive platelets were detected in IBD-involved mucosa. Activated platelets up-regulated expression of intercellular adhesion molecule 1 and vascular cell adhesion molecule 1 as well as production of interleukin 8 by HIMEC in a CD40-dependent fashion. High levels of RANTES were present in platelet-HIMEC cocultures and platelets were identified as the source of this chemokine, which mediated T-cell adhesion to HIMEC. CONCLUSIONS: These results show that platelets can actively contribute to mucosal inflammation and represent a previously unrecognized component of IBD pathogenesis. PMID- 12730867 TI - Impaired expression of peroxisome proliferator-activated receptor gamma in ulcerative colitis. AB - BACKGROUND & AIMS: The peroxisome proliferator-activated receptor gamma (PPAR gamma) has been proposed as a key inhibitor of colitis through attenuation of nuclear factor kappa B (NF-kappa B) activity. In inflammatory bowel disease, activators of NF-kappa B, including the bacterial receptor toll-like receptor (TLR)4, are elevated. We aimed to determine the role of bacteria and their signaling effects on PPAR gamma regulation during inflammatory bowel disease (IBD). METHODS: TLR4-transfected Caco-2 cells, germ-free mice, and mice devoid of functional TLR4 (Lps(d)/Lps(d) mice) were assessed for their expression of PPAR gamma in colonic tissues in the presence or absence of bacteria. This nuclear receptor expression and the polymorphisms of gene also were assessed in patients with Crohn's disease (CD) and ulcerative colitis (UC), 2 inflammatory bowel diseases resulting from an abnormal immune response to bacterial antigens. RESULTS: TLR4-transfected Caco-2 cells showed that the TLR4 signaling pathway elevated PPAR gamma expression and a PPAR gamma-dependent reporter in an I kappa kappa beta dependent fashion. Murine and human intestinal flora induced PPAR gamma expression in colonic epithelial cells of control mice. PPAR gamma expression was significantly higher in the colon of control compared with Lps(d)/Lps(d) mice. Although PPAR gamma levels appeared normal in patients with CD and controls, UC patients displayed a reduced expression of PPAR gamma confined to colonic epithelial cells, without any mutation in the PPAR gamma gene. CONCLUSIONS: These data showed that the commensal intestinal flora affects the expression of PPAR gamma and that PPAR gamma expression is considerably impaired in patients with UC. PMID- 12730868 TI - Emergency sclerotherapy versus vasoactive drugs for variceal bleeding in cirrhosis: a Cochrane meta-analysis. AB - BACKGROUND & AIMS: Emergency sclerotherapy is used as a first-line therapy for variceal bleeding in cirrhosis, although pharmacologic treatment stops bleeding in most patients. We performed a meta-analysis comparing emergency sclerotherapy with pharmacologic treatment. METHODS: MEDLINE (1968-2002), EMBASE (1986-2002), and the Cochrane Library (2002;4) were searched to retrieve randomized controlled trials comparing sclerotherapy with vasopressin (+/- nitroglycerin), terlipressin, somatostatin, or octreotide for variceal bleeding in cirrhosis. Outcome measures were failure to control bleeding, rebleeding, blood transfusions, adverse events, and mortality. RESULTS: Fifteen trials were identified. Sclerotherapy was not superior to terlipressin, somatostatin, or octreotide for any outcome and to vasopressin for rebleeding, blood transfusions, death, and adverse events; it was superior to vasopressin for the control of bleeding in a single trial flawed by a potential detection bias. Sclerotherapy was associated with significantly more adverse events than somatostatin. In a predefined sensitivity analysis, combining all of the trials irrespective of the control treatment, risk differences (sclerotherapy minus control) and confidence intervals (CIs) were as follows: failure to control bleeding, -0.03 (-0.06 to 0.01); mortality, -0.035 (-0.07 to 0.008); adverse events, 0.08 (0.02 to 0.14). Mortality risk difference was -0.01 (-0.07 to 0.04) in good-quality trials and 0.08 (-0.14 to -0.02) in poor-quality trials. CONCLUSIONS: Available evidence does not support emergency sclerotherapy as the first-line treatment of variceal bleeding in cirrhosis when compared with vasoactive drugs, which control bleeding in 83% of patients. Therefore, endoscopic therapy might be added only in pharmacologic treatment failures. PMID- 12730869 TI - Risk factors for the development of pancreatic cancer in familial pancreatic cancer kindreds. AB - BACKGROUND & AIMS: Approximately 10% of pancreatic cancers are inherited, but the factors that affect tumorigenesis in familial pancreatic cancer are unknown. We sought to determine whether smoking or other factors could predict cancer risk in familial pancreatic cancer kindreds. METHODS: We conducted a nested case-control study including 251 members of 28 families. All families included 2 or more members with pancreatic cancer. We determined the effects of smoking, young age of onset within the family, diabetes mellitus, sex, and number/standing of affected relatives on the risk of pancreatic cancer. RESULTS: Smoking was an independent risk factor for familial pancreatic cancer (odds ratio [OR], 3.7; 95% confidence interval [CI], 1.8-7.6), and the risk was greatest in males and subjects younger than 50 (OR, 5.2 and OR, 7.6, respectively). Smokers developed cancer 1 decade earlier than nonsmokers (59.6 vs. 69.1 years; P = 0.01), and the number of affected first-degree relatives also increased risk (OR, 1.4; 95% CI, 1.1-1.9 for each additional family member). Diabetes was not a risk factor for pancreatic cancer, although diabetes was associated with pancreatic dysplasia. One third of families demonstrated genetic anticipation, as the mean age of onset decreased by 2 decades between generations. CONCLUSIONS: Smoking is a strong risk factor in familial pancreatic cancer kindreds, particularly among males and those under age 50. Persons with multiple affected first-degree relatives are also at increased risk. These factors may be useful in selecting candidates for pancreatic cancer screening. Members of families with multiple pancreatic cancers should be counseled not to smoke. PMID- 12730871 TI - Frequent mutations of hepatocyte nuclear factor 1 in colorectal cancer with microsatellite instability. AB - BACKGROUND & AIMS: The TCF1 gene encoding hepatocyte nuclear factor 1 alpha (HNF1), a transcription factor germline mutated in patients with maturity-onset diabetes of the young type 3, was recently found to be frequently inactivated by biallelic alterations in liver adenoma and in rare hepatocellular carcinomas. The impact of HNF1 in colorectal carcinogenesis has not been studied until now. Colorectal cancer is characterized by the existence of different molecular mechanisms known as microsatellite stable or unstable tumors. METHODS: At first, a series of 10 adenomas and 29 colon cancers regardless of microsatellite instability status were screened for TCF1 mutations on the entire coding sequence. RESULTS: Three mutations in microsatellite instability high (MSI-H) tumors were found in the exon 4 polymorphic poly-cytosin (C)(8) or (C)(9) tract and consisted of a cytosin deletion at position 291. To further characterize the prevalence of TCF1 mutations in the subgroup of MSI-H tumors, 52 additional MSI-H samples were screened for exon 4 alterations; 23% of MSI-H tumors (95% confidence interval, 14%-36%) were found to harbor frameshift at the poly-cytosin tract. The (C)(9) allele was significantly more frequently mutated than the (C)(8) allele (22% vs. 8%; P = 0.03), showing a higher instability of the longer repetition. CONCLUSIONS: These results show a role for HNF1 in MSI-H colorectal carcinogenesis. PMID- 12730870 TI - Epigenetic and genetic alterations in duodenal carcinomas are distinct from biliary and ampullary carcinomas. AB - BACKGROUND & AIMS: Carcinomas of the extrahepatic bile ducts, ampulla of Vater, and duodenum are uncommon, and their epigenetic and genetic alterations are not well characterized. METHODS: We therefore compared the methylation profile and genetic alterations in 18 extrahepatic biliary, 9 ampullary, and 12 duodenal carcinomas. We evaluated methylation at p16, p14, and human Mut L homologue (hMLH1) by methylation- specific PCR (MSP), and at cyclooxygenase 2 (COX2), O(6) methyl-guanine methyltransferase (MGMT), estrogen receptor (ER), retinoic acid receptor beta 2 (RAR beta), and T-type calcium channel (CACNA1G) genes, and methylated in tumor 1 (MINT1), MINT2, MINT25, MINT27, and MINT31 loci by combined bisulfite restriction analysis (COBRA); mutation of K-ras, p53, p16, and p14 genes by sequencing; loss of heterozygosity of chromosome 9p; and microsatellite instability (MSI). RESULTS: Duodenal carcinomas were methylated more frequently or had increased methylation densities than biliary carcinomas at p14 (P = 0.04), hMLH1 (P = 0.04), MGMT (P = 0.01), MINT1 (P = 0.01), MINT25 (P = 0.01), MINT27 (P = 0.001), RAR beta (P = 0.03), and ER (P = 0.001), and than ampullary carcinomas at RAR beta (P = 0.02) and ER (P = 0.03). In contrast, the methylation profiles of biliary and ampullary carcinomas were not statistically different. Simultaneous methylation of 3 or more CpG islands (CpG island methylator phenotype-high) was more common in duodenal cancers (P = 0.004). MGMT methylation was associated with G-to-A mutation in K-ras (P = 0.006), and hMLH1 methylation was associated with MSI-high (P = 0.01). CONCLUSIONS: Our findings indicate that the methylation profile and genetic alterations of duodenal carcinomas are distinct from biliary and ampullary carcinomas, and that tumor-specific methylation is associated with gene mutation and MSI. PMID- 12730872 TI - A novel PPAR gamma gene therapy to control inflammation associated with inflammatory bowel disease in a murine model. AB - BACKGROUND & AIMS: Peroxisome proliferator-activated receptor gamma (PPAR gamma) is one of the nuclear receptors that plays a central role in adipocyte differentiation and insulin sensitivity. PPAR gamma has also recently been recognized as an endogenous regulator of intestinal inflammation. However, its levels are decreased during chronic inflammation in human and mice, thus limiting PPAR gamma ligand therapy during established disease. We sought to determine whether this decrease in PPAR gamma could be counteracted by a gene therapy approach. METHODS: We characterized PPAR gamma levels in experimental colitis associated with dextran sodium sulfate administration to mice. In this model, the therapeutic benefits of PPAR gamma gene therapy using a replication-deficient adenovirus vector expressing PPAR gamma (Ad-PPAR gamma) was assessed. RESULTS: PPAR gamma protein levels were decreased in whole colonic tissue, lamina propria lymphocytes, and peritoneal exudate cells during the course of colitis. PPAR gamma gene delivery using Ad-PPAR gamma restored responsiveness to a PPAR gamma ligand, resulting in marked amelioration of tissue inflammation associated with the colitis, which included attenuation of intercellular adhesion molecule-1, cyclooxygenase-2 and tumor necrosis factor-alpha expression. CONCLUSIONS: Our results suggest that gene delivery of PPAR gamma can be used to restore and/or enhance endogenous anti-inflammatory processes that are normally operative in mammalian tissues such as in the colon. PMID- 12730874 TI - Epidermal growth factor receptor-related protein: a potential therapeutic agent for colorectal cancer. AB - BACKGROUND & AIMS: Epidermal growth factor receptor is frequently implicated in epithelial cancers and is, therefore, being considered as a potential target for therapy. Recently, we reported the isolation and characterization of epidermal growth factor receptor-related protein, a negative regulator of epidermal growth factor receptor. To discern whether epidermal growth factor receptor-related protein could be an effective therapeutic agent for colorectal cancer, we generated epidermal growth factor receptor-related protein fusion protein and studied its effect on the growth of colon cancer cells in vivo and in vitro. We also studied whether epidermal growth factor receptor-related protein expression is altered in colorectal cancer. METHODS: A 55-kilodalton epidermal growth factor receptor-related protein fusion protein with V5 and His tags was generated in a drosophila expression system and subsequently purified by a His antibody affinity column. Rabbit polyclonal antibodies against epidermal growth factor receptor related protein were used to examine the expression of epidermal growth factor receptor-related protein. RESULTS: Epidermal growth factor receptor-related protein expression was found to be high in benign human colonic epithelium but low in adenocarcinoma. Exposure of the colon cancer cell lines HCT-116 and Caco-2 to purified recombinant epidermal growth factor receptor-related protein caused a marked inhibition of proliferation, as well as attenuation of basal and ligand induced stimulation of epidermal growth factor receptor phosphorylation. Epidermal growth factor receptor-related protein-induced inhibition of proliferation of colon cancer cells was prevented by epidermal growth factor receptor-related protein antibodies. Reduced epidermal growth factor receptor phosphorylation was partly due to sequestration of epidermal growth factor receptor ligands by epidermal growth factor receptor-related protein, resulting in the formation of inactive heterodimers with epidermal growth factor receptor. Intratumoral or subcutaneous (away from the tumor site) injections of purified epidermal growth factor receptor-related protein caused regression of palpable colon cancer xenograft tumors in some severely compromised immunodeficient mice and arrested tumor growth in others. CONCLUSIONS: We propose that epidermal growth factor receptor-related protein inhibits cellular growth by attenuating epidermal growth factor receptor signaling processes and is an effective therapeutic agent for colorectal cancer. PMID- 12730873 TI - Characterization of the effects of pancreatic polypeptide in the regulation of energy balance. AB - BACKGROUND & AIMS: Pancreatic polypeptide (PP) belongs to a family of peptides including neuropeptide Y and peptide YY. We examined the role of PP in the regulation of body weight as well as the therapeutic potential of PP. METHODS: We measured food intake, gastric emptying, oxygen consumption, and gene expression of hypothalamic neuropeptides, gastric ghrelin, and adipocytokines in mice after administering PP intraperitoneally. Peptide gene expression was also examined in PP-overexpressing mice. Vagal and sympathetic nerve activities were recorded after intravenous administration in rats. Effects of repeated administrations of PP on energy balance and on glucose and lipid metabolism were examined in both ob/ob obese mice and fatty liver Shionogi (FLS)-ob/ob obese mice. RESULTS: Peripherally administered PP induced negative energy balance by decreasing food intake and gastric emptying while increasing energy expenditure. The mechanism involved modification of expression of feeding-regulatory peptides (decrease in orexigenic neuropeptide Y, orexin, and ghrelin along with an increase in anorexigenic urocortin) and activity of the vagovagal or vagosympathetic reflex arc. PP reduced leptin in white adipose tissue and corticotropin-releasing factor gene expression. The expression of gastric ghrelin and hypothalamic orexin was decreased in PP-overexpressing mice. Repeated administrations of PP decreased body weight gain and ameliorated insulin resistance and hyperlipidemia in both ob/ob obese mice and FLS-ob/ob obese mice. Liver enzyme abnormalities in FLS ob/ob obese mice were also ameliorated by PP. CONCLUSIONS: These observations indicate that PP may influence food intake, energy metabolism, and the expression of hypothalamic peptides and gastric ghrelin. PMID- 12730875 TI - Progastrin stimulates murine colonic epithelial mitosis after DNA damage. AB - BACKGROUND & AIMS: Transgenic mice that overexpress progastrin are more susceptible than either wild-type mice or mice that overexpress amidated gastrin to chemical carcinogen-induced colonic adenomas. We have investigated whether alterations in the regulation of apoptosis or mitosis after DNA damage contribute to the effects of progastrin on murine colonic epithelium. METHODS: Apoptosis and mitosis were assessed on a cell positional basis in murine intestinal epithelium after gamma-irradiation. Mice analyzed were progastrin overexpressing, gastrin overexpressing, gastrin knockout, and their wild-type counterparts. The expression of cell cycle regulators was analyzed by gene array and Western blotting. RESULTS: Apoptosis was induced to similar levels in the small intestinal and colonic crypts of all mice 4.5 hours after 8 Gy gamma-radiation. Colonic mitosis was inhibited to almost undetectable levels by 8Gy gamma radiation in wild-type, gastrin-knockout, and gastrin-overexpressing mice. However, significant colonic mitosis persisted in progastrin-overexpressing mice up to 24 hours after 8Gy gamma-radiation. Increased levels of cdk4 and cyclin D1 proteins were found in the colonic epithelium of progastrin-overexpressing mice relative to wild-type animals after gamma-radiation. CONCLUSIONS: After DNA damage by gamma-radiation, mice with elevated progastrin exhibit significantly higher levels of colonic mitosis than wild-type or gastrin-overexpressing mice. Persistently elevated cdk4 and cyclin D1 in progastrin overexpressing mice accounts for the capacity of colon cells to continue with the cell cycle after DNA damage. PMID- 12730876 TI - Interleukin-11-induced heat shock protein 25 confers intestinal epithelial specific cytoprotection from oxidant stress. AB - BACKGROUND & AIMS: The mechanisms of interleukin-11 (IL-11) cytoprotection in intestinal epithelial injury are largely unknown. IL-11 protects barrier integrity during oxidant stress, a common endpoint of numerous types of intestinal injury including ischemia and immune-mediated inflammation. Because heat shock proteins (hsp) are cytoprotective in intestinal epithelia, we hypothesized that IL-11-conferred cytoprotection is mediated by inducible hsps. METHODS: IL-11 receptor (IL-11R) activation was determined using phospho-specific antibodies to STAT3. IL-11 induction of hsp72 and hsp25 was determined by immunoblot in IEC-18 crypt and young adult mouse colon colonic epithelial cells. Epithelial resistance to oxidant injury by monochloramine was determined by (51)Cr release. Stable hsp anti-sense IEC-18 cell clones were obtained by electroporation and hygromycin B selection. The IL-11 effect on hsp25 distribution was characterized by analysis of Triton x-100 insoluble fractions, 2 D isoelectric focusing gels, and confocal microscopy. RESULTS: IL-11R signaling was detected in all cells under study. IL-11 induces hsp25 in an intestinal epithelial-specific manner that significantly preserves cellular viability in the presence of monochloramine. This effect was significantly reversed in intestinal epithelia stably expressing anti-sense to hsp25. IL-11 induced a shift of hsp25 to Triton x-100 insoluble fractions containing cytoskeletal elements, which was not associated with altered hsp25 phosphorylation. The shift was not paralleled by increased hsp25 co-localization with F-actin by confocal microscopy. CONCLUSIONS: The induction of hsp25 by IL-11 confers epithelial-specific cytoprotection that is independent of phosphorylation-dependent co-localization of hsp25 to F-actin, thereby contributing to the protective effects of IL-11 in models of intestinal epithelial injury. PMID- 12730877 TI - NF-kappa B activation by oxidative stress and inflammation suppresses contractility in colonic circular smooth muscle cells. AB - BACKGROUND & AIMS: Transcription factor nuclear factor kappa B (NF-kappa B) plays a critical role in transcriptional changes in several diseases, including inflammation. The aim of this study was to investigate whether NF-kappa B is activated by inflammation and oxidative stress in colonic circular smooth muscle cells and whether that leads to suppression of their contractility. METHODS: The experiments were performed on freshly dissociated single cells using electrophoretic mobility shift assay, Western immunoblotting, and immunofluorescence imaging. RESULTS: The NF-kappa B DNA binding was approximately 6-fold greater in cells from the inflamed colon vs. those from the normal colon. Supershift assay indicated that the antibodies to p65, p50, and c-Rel, but not that to p52, shifted the NF-kappa B band. Western immunoblotting and immunofluorescence imaging also demonstrated the presence of p65, p50, and c-Rel proteins in the cytoplasm and their translocation to the nucleus by H(2)O(2) induced oxidative stress. H(2)O(2) treatment degraded I kappa B(beta), but not I kappa B(alpha), to translocate NF-kappa B to the nucleus. Hydrogen peroxide concentration and time dependently activated NF-kappa B DNA binding and suppressed cell contraction to acetylcholine. NF-kappa B inhibitors significantly inhibited these effects. Inhibition of NF-kappa B prior to and during inflammation in intact dogs also reversed the suppression of contractility. CONCLUSIONS: Transcription factor NF-kappa B is activated in colonic circular muscle cells by inflammation and oxidative stress. This activation of NF-kappa B mediates the suppression of cell contractility. PMID- 12730878 TI - Galectin-1 suppresses experimental colitis in mice. AB - BACKGROUND & AIMS: Uncontrolled T-cell activation plays a critical role in the pathogenesis of inflammatory bowel diseases. Therefore, pharmacologic strategies directed to restore the normal responsiveness of the immune system by deleting inappropriately activated T cells could be efficacious in the treatment of these pathologic conditions. Galectin-1 is an endogenous lectin expressed in lymphoid organs that plays a role in the maintenance of central and peripheral tolerance. The aim of the present study was to evaluate the therapeutic effects of galectin 1 on T-helper cell type 1-mediated experimental colitis induced by intrarectal administration of 2,4,6-trinitrobenzene sulfonic acid (TNBS) in mice. METHODS: Cells and tissues from mice with TNBS colitis receiving treatment with several doses of human recombinant galectin-1 (hrGAL-1) were analyzed for morphology, cytokine production, and apoptosis. RESULTS: Prophylactic and therapeutic administration of rhGAL-1 resulted in a striking improvement in the clinical and histopathologic aspects of the disease. hrGAL-1 reduced the number of hapten activated spleen T cells, decreased inflammatory cytokine production, and profoundly reduced the ability of lamina propria T cells to produce IFN gamma in vitro. Moreover, hrGAL-1 led to the appearance of apoptotic mononuclear cells in colon tissue when administered in vivo and induced selective apoptosis of TNBS activated lamina propria T cells in vitro. CONCLUSION: Collectively, these data show that hrGAL-1 exerts protective and immunomodulatory activity in TNBS-induced colitis and it might be effective in the treatment of inflammatory bowel diseases. PMID- 12730879 TI - Enteric flora and lymphocyte-derived cytokines determine expression of heat shock proteins in mouse colonic epithelial cells. AB - BACKGROUND & AIMS: Inducible heat shock proteins (hsps), particularly hsp25 and hsp72, are expressed by surface colonocytes and may have a role in protecting intestinal epithelial cells against injury. This study is aimed at determining if enteric bacteria and/or immune signals regulate their physiologic expression. METHODS: Intestinal hsp25, hsp72, and constitutive hsc73 expression were studied in immunodeficient RAG-1(-/-) mice and in normal mice. Mucosal permeability was measured by mannitol flux and transepithelial resistance. Hsp expression in intestinal YAMC cells was assessed after incubation with recombinant cytokines, activated lamina propria lymphocytes (LPLs), or Bacteroides fragilis. RESULTS: Chronic metronidazole treatment decreases colonic mucosal hsp25 and hsp72 expression, an effect associated with increased susceptibility of mucosal barrier function to C. difficile toxin A. Hsp expression also was increased in YAMC cells incubated with B. fragilis, an effect mediated by lipopolysaccharide and other bacteria-derived factors. Colonic hsp72, but not hsp25 or hsc73, expression is decreased in RAG-1(-/-) mice. Recombinant IL-2 and other cytokines enhance YAMC hsp25 and/or hsp72 expression. Activated LPLs induce YAMC hsp expression, an effect blocked by IL-2 neutralizing antibody. CONCLUSIONS: Enteric flora and mucosal lymphocytes play a role in maintaining physiologic expression of colonocyte hsp25 and hsp72. PMID- 12730880 TI - Chronic Helicobacter pylori infections induce gastric mutations in mice. AB - BACKGROUND & AIMS: Helicobacter pylori is an important etiologic factor in the development of gastric cancer. The aim of this study was to analyze the role of H. pylori infections in the induction of mutagenic events in gastric epithelial cells. The effect of a high-salt diet as a genotoxic risk factor was also investigated. METHODS: Big Blue transgenic male mice (C57Bl/6) were inoculated with H. pylori (strain SS1) or Helicobacter felis (strain CS1) for 6 and 12 months. The frequency and spectrum of mutations at the stomach level were assessed. Inflammatory host response and inducible nitric oxide synthase (iNOS) expression by reverse-transcription polymerase chain reaction and immunohistochemistry analysis were also performed. RESULTS: After 6 months, the gastric mutant frequency was 4-fold and 1.7-fold higher in mice infected with H. pylori and H. felis, respectively, than in uninfected mice. It was associated with a high frequency of transversions (AT --> CG and GC --> TA) known to result from oxidative damages. The Helicobacter-infected mice exhibited severe gastritis and a high level of iNOS messenger RNA expression. Hyperplasia developed 12 months after inoculation, and both the mutagenic effects and iNOS expression decreased in H. pylori- and H. felis-infected mice. No synergistic effects of a high-salt diet and Helicobacter infection were observed regarding the frequency of gastric mutation. CONCLUSIONS: A direct gastric mutagenic effect due to H. pylori infection in the Big Blue transgenic mouse model has been shown 6 months after inoculation. This genotoxicity can be attributable to oxidative DNA damage involving the inflammatory host response. PMID- 12730881 TI - Delineation of a CD1d-restricted antigen presentation pathway associated with human and mouse intestinal epithelial cells. AB - BACKGROUND & AIMS: CD1d, a major histocompatibility complex (MHC) class I-related molecule that is responsible for the presentation of glycolipid antigens to subsets of natural killer T (NK-T) cells, is expressed by intestinal epithelial cells (IECs). However, CD1d-restricted antigen presentation has not yet been examined on IECs. METHODS: A mouse intestinal epithelial cell line (MODE-K), a human epithelial cell line (T84), T84 cells transfected with CD1d and/or MHC class II, and freshly isolated human IECs were examined for their ability to present model glycolipid antigens to NK-T cells as defined by interleukin (IL)-2 or IL-4 secretion. RESULTS: MODE-K and freshly isolated human IECs exhibited dose dependent, CD1d-restricted presentation of the functional glycolipid antigen, alpha-galactosylceramide (alpha GalCer), to the mouse NK-T cell hybridoma, DN32.D3. The human IEC line, T84, mainly presented alpha GalCer when transfected with human CD1d. Presentation of alpha GalCer by CD1d-transfected T84 cells (T84d) to DN32.D3 cells was greater along the basal surface in comparison with the apical surface. Induction of the MHC class II antigen presentation machinery by cotransfecting T84d with the MHC class I transactivator (CIITA) did not alter this polarity of presentation. Neither MODE-K nor T84 cells transfected with CD1d, CD1d plus CIITA, or CD1d plus HLA-DR were able to present glycolipid antigens requiring intracellular processing. The MODE-K cell line could also present alpha GalCer to primary mouse NK-T cells. CONCLUSIONS: CD1d is expressed functionally on IECs with a polarity of presentation (basal > apical) predicting a role in presentation of mucosal glycolipid antigens to local CD1d-restricted T cells. PMID- 12730882 TI - Localization of the ammonium transporters, Rh B glycoprotein and Rh C glycoprotein, in the mouse liver. AB - BACKGROUND & AIMS: Hepatic ammonium metabolism is critical for maintenance of normal health. Three mammalian members of an ammonium transporter family have recently been identified: Rh A glycoprotein (RhAG), Rh B glycoprotein (RhBG), and Rh C glycoprotein (RhCG). This study examined which of these are expressed in the mouse liver and in which cells they are expressed. METHODS: Normal Balb/c mice were used. Messenger RNA (mRNA) expression was detected using either conventional or real-time reverse-transcription polymerase chain reaction (RT-PCR). Protein expression was examined using immunoblot analysis and either immunohistochemical or immunofluorescent microscopy. RESULTS: We confirmed hepatic RhBG mRNA expression using real-time RT-PCR. Immunoblot analysis identified expression of a approximately 45-kilodalton protein. Immunohistochemical and immunofluorescent microscopy identified basolateral RhBG immunoreactivity in 1-2 cell layers of hepatocytes surrounding central veins. No immunoreactivity was identified in periportal or midzonal hepatocytes. Perivenous hepatocyte-specific expression was confirmed by colocalization with glutamine synthetase. A second ammonium transporter, RhCG, was expressed but at substantially lower levels. Real-time RT PCR quantified hepatic RhCG mRNA expression at approximately 0.4% of RhBG mRNA expression. Immunoblot analysis confirmed RhCG protein expression, and immunofluorescence microscopy identified RhCG expression in bile duct epithelia. In contrast to RhBG and RhCG, RhAG mRNA was not identified by RT-PCR. CONCLUSIONS: RhBG and RhCG are expressed by the mouse liver. Basolateral RhBG is expressed by perivenous hepatocytes, where it may mediate ammonium uptake, and RhCG immunoreactivity is present in bile duct epithelial cells, where it may contribute to ammonium secretion into bile fluid. PMID- 12730883 TI - IFN alpha-2a protects mice against a helminth infection of the liver and modulates immune responses. AB - BACKGROUND & AIMS: Hepatic alveolar echinococcosis (AE), caused by the larval growth of Echinococcus multilocularis, is one of the most lethal helminthic diseases with no satisfactory treatment. Advances in the understanding of the host's immune response (Th2 responses associated with a progressive form of AE), have driven the research towards immune stimulation as an alternative possibility to treat patients. We previously reported clinical stabilization associated with a shift from a Th2 to a Th1 cytokine profile in an AE patient treated with interferon (IFN)alpha. METHODS: The effects of recombinant IFN alpha-2a were analyzed in the susceptible C57BL/6J E. multilocularis infected mice. Parasitic burden, macrophage functions, and specific T-cell responses were studied 15, 45, and 90 days postinfection. RESULTS: After 90 days postinfection, 75% of infected IFN alpha-2a-treated mice had no hepatic lesions and half were fully protected. IFN alpha-2a treatment markedly decreased the abnormally elevated production of IL-10 in both spleen cell cultures and peritoneal macrophage cultures from infected mice and restored phagocytosis and oxidative metabolism of macrophages. It also inhibited IL-6 and IL-13 antigen-induced secretions in spleen cell cultures. CONCLUSIONS: Through its immunoregulatory properties, IFN alpha-2a may be effective in a helminthic liver infection and is a promising candidate for clinical application in AE. PMID- 12730884 TI - Leptin-specific mechanisms for impaired liver regeneration in ob/ob mice after toxic injury. AB - BACKGROUND & AIMS: Profound impairment of liver regeneration in rodents with dysfunctional leptin signaling has been attributed to non-alcohol-induced fatty liver disorders (NAFLD). Our aim was to establish whether defective liver regeneration in ob/ob mice is a direct consequence of leptin-dependent, intracellular signaling mechanisms controlling cell-cycle regulation in hepatocytes. METHODS: After exposure to a single hepatotoxic dose of (CCl(4)), the regenerative response to hepatic injury was studied in leptin-deficient ob/ob and control mice. The effects of leptin supplementation (100 microg x kg(-1) x day(-1)) were examined. We assessed entry into and progression through the cell cycle and activation of key signaling intermediates and transcriptional regulators. RESULTS: CCl(4)-induced liver injury was equally severe in ob/ob and control mice. In leptin-deficient mice, it was associated with exaggerated activation of NF-kappa B and STAT3 during the priming phase, abrogation of tumor necrosis factor (TNF) and interleukin (IL)-6 release at the time of G1/S transition, and failure of hepatocyte induction of cyclin D1 and cell cycle entry. Leptin replacement corrected these defects in ob/ob mice by restoring TNF and IL-6 release and inducing cyclin D1. Hepatocytes entered S phase and progressed, as in wild-type mice, to vigorous mitosis and normal hepatic regenerative response. In ob/ob mice, low doses of TNF before CCl(4) also were associated with restitution of TNF release and proliferative capabilities. CONCLUSIONS: Impaired liver regeneration in ob/ob mice is caused by leptin deficiency. We propose that altered cytokine production in ob/ob mice is part of the mechanisms responsible for impaired proliferation in response to hepatic injury. PMID- 12730885 TI - Expression of hepatitis c virus proteins inhibits interferon alpha signaling in the liver of transgenic mice. AB - BACKGROUND & AIMS Hepatitis C virus (HCV) is a major cause of chronic liver disease, cirrhosis, and hepatocellular carcinoma worldwide. The majority of patients treated with interferon alpha do not have a sustained response with clearance of the virus. The molecular mechanisms underlying interferon resistance are poorly understood. Interferon-induced activation of the Jak-STAT (signal transducer and activator of transcription) signal transduction pathway is essential for the induction of an antiviral state. Interference of viral proteins with the Jak-STAT pathway could be responsible for interferon resistance in patients with chronic HCV. METHODS: We have analyzed interferon-induced signal transduction through the Jak-STAT pathway in transgenic mice that express HCV proteins in their liver cells. STAT activation was investigated with Western blots, immunofluorescence, and electrophoretic mobility shift assays. Virus challenge experiments with lymphocytic choriomeningitis virus were used to demonstrate the functional importance of Jak-STAT inhibition. RESULTS: STAT signaling was found to be strongly inhibited in liver cells of HCV transgenic mice. The inhibition occurred in the nucleus and blocked binding of STAT transcription factors to the promoters of interferon-stimulated genes. Tyrosine phosphorylation of STAT proteins by Janus kinases at the interferon receptor was not inhibited. This lack in interferon response resulted in an enhanced susceptibility of the transgenic mice to infection with a hepatotropic strain of lymphocytic choriomeningitis virus. CONCLUSIONS: Interferon-induced intracellular signaling is impaired in HCV transgenic mice. Interference of HCV proteins with interferon-induced intracellular signaling could be an important mechanism of viral persistence and treatment resistance. PMID- 12730886 TI - Involvement of integrins and Src in tauroursodeoxycholate-induced and swelling induced choleresis. AB - BACKGROUND & AIMS: Stimulation of canalicular secretion by tauroursodeoxycholate (TUDC) involves dual activation of p38 mitogen-activated protein kinase (p38(MAPK)) and extracellular signal-regulated kinase (ERK). This study investigates the sensing and upstream signaling events of TUDC-induced choleresis. METHODS: TUDC and hypo-osmolarity effects on protein kinase activities and taurocholate excretion were studied in perfused rat liver. RESULTS: TUDC induced a rapid activation of focal adhesion kinase (FAK) and Src, as shown by an increase in Y418 phosphorylation and a decrease in Y529 phosphorylation of Src. Inhibition of Src by PP-2 abolished the TUDC-induced activation of p38(MAPK) but not of FAK and ERKs. An integrin-inhibitory peptide with an RGD motif blocked TUDC-induced FAK, Src, ERK, and p38(MAPK) activation, suggesting that integrin signaling toward FAK/Src is required for TUDC-induced MAPK activation. The RGD peptide and PP-2 also abolished the stimulation of taurocholate excretion in perfused rat liver in response to TUDC. Integrin dependent Src activation was also identified as an upstream event in hypo-osmotic signaling toward MAPKs and choleresis. CONCLUSIONS: TUDC-induced stimulation of canalicular taurocholate excretion involves integrin sensing, FAK, and Src activation as upstream events for dual MAPK activation. Integrins may also represent one long-searched sensor for cell hydration changes in response to hypo osmolarity. PMID- 12730887 TI - Betaine decreases hyperhomocysteinemia, endoplasmic reticulum stress, and liver injury in alcohol-fed mice. AB - BACKGROUND & AIMS: Alcohol-induced hyperhomocysteinemia has been reported in rats and humans. Hyperhomocysteinemia has been associated with endoplasmic reticulum (ER) stress leading to the activation of ER-dependent apoptosis or up-regulation of lipid synthesis. This novel ER stress mechanism of alcoholic liver injury was studied in the model of intragastric alcohol-fed mice. METHODS: Effects of alcohol on gene expression were analyzed using cDNA microarrays, RT-PCR, and Western blots over a period of 6 weeks. Liver injury was examined by histologic staining and TUNEL. RESULTS: We observed fatty liver, increased hepatic necroinflammation and apoptosis, and hyperhomocysteinemia. Of 1176 toxicology related genes, glucose-regulated proteins (GRP-78 and -94), growth arrest/DNA damage-inducible protein 153 (CHOP/GADD153), and caspase-12 indicative of an ER stress response were among the alcohol-responsive genes. Sterol regulatory element binding protein (SREBP-1) and HMG-CoA reductase also were enhanced with alcohol administration. RT-PCR and selective Western blots confirmed the alcohol induced expression of ER stress-related apoptosis and lipid synthesis genes. Addition of 0.5% and maximal 1.5% betaine to the alcohol diet reduced the elevated level of plasma homocysteine by 54% and more than 80% accompanied by a decrease in hepatic lipids and ER stress response. Betaine did not attenuate the ethanol-induced increase in tumor necrosis factor alpha or CD14 mRNA. CONCLUSIONS: The results strongly suggest that alcohol may modulate both apoptotic and fat synthetic gene expression through homocysteine-induced ER stress in chronic alcoholic mouse liver and that correction of hyperhomocysteinemia by betaine or other approaches may be useful to prevent alcoholic liver disease. PMID- 12730888 TI - The role of nitric oxide synthase isoforms in extrahepatic portal hypertension: studies in gene-knockout mice. AB - BACKGROUND & AIMS: Considerable debate exists concerning which isoform of nitric oxide synthase (NOS) is responsible for the increased production of NO in PHT. We used the portal vein ligation model of PHT in wild-type and eNOS- or iNOS knockout mice to definitively determine the contribution of these isoforms in the development of PHT. METHODS: The portal vein of wild-type mice, or those with targeted mutations in the nos2 gene (iNOS) or the nos3 gene (eNOS), was ligated and portal venous pressure (Ppv), abdominal aortic blood flow (Qao), and portosystemic shunt determined 2 weeks later. RESULTS: In wild-type mice, as compared with sham-operated controls, portal vein ligation (PVL) resulted in a time-dependent increase in Ppv (7.72 +/- 0.37 vs 17.57 +/- 0.51 cmH(2)O, at 14 days) concomitant with a significant increase in Qao (0.12 +/- 0.003 vs 0.227 +/- 0.005 mL/min/g) and portosystemic shunt (0.47% +/- 0.01% vs 84.13% +/- 0.09% shunt). Likewise, PVL in iNOS-deficient mice resulted in similar increases in Ppv, Qao, and shunt development. In contrast, after PVL in eNOS-deficient animals, there was no significant change in Ppv (7.52 +/- 0.22 vs 8.07 +/- 0.4 cmH(2)0) or Qao (0.111 +/- 0.01 vs 0.14 +/-.023 mL/min/g). However, eNOS (-/-) mice did develop a substantial portosystemic shunt (0.33% +/- 0.005% vs 84.53% +/ 0.19% shunt), comparable to that seen in wild-type animals after PVL. CONCLUSIONS: These data support a key role for eNOS, rather than iNOS, in the pathogenesis of PHT. PMID- 12730889 TI - Hemochromatosis gene modifies course of hepatitis C viral infection. PMID- 12730890 TI - Our new president--Daniel K. Podolsky, M.D. PMID- 12730891 TI - Ghrelin-leptin tango in body-weight regulation. PMID- 12730892 TI - Prevention is the best defense: Probiotic prophylaxis of pouchitis. PMID- 12730893 TI - Is there a role for PPAR gamma in IBD? Yes, no, maybe. PMID- 12730917 TI - Proceedings of the 2002 Current Concepts in Joint Replacement meeting. Las Vegas, Nevada, USA. PMID- 12730894 TI - Pancreatic polypeptide: more than just another gut hormone? PMID- 12730918 TI - Lessons from the other side of the knife. AB - The overall success or failure of knee arthroplasty is predicated on the successful combination of patient selection, meticulous surgical technique, and prosthetic design. Discussions regarding the outcome of knee arthroplasty frequently focus on the implants used and the surgical technique employed, with a less frequent focus on patient profile. However, the patient experience was further highlighted to me when I became the patient and experienced the consequences of knee arthroplasty firsthand. This experience offered me a greater insight into patient expectations, the importance of preoperative counseling, the severity of perioperative pain, and the issues involved in the rehabilitative period. Joint arthroplasty outcomes are dependent on the harmony existing between the patient's perioperative status, expectation level, motivational status, physician diagnostic and surgical skill, and the design characteristics of the prosthesis. PMID- 12730919 TI - Glucosamine and chondroitin sulfate are effective in the management of osteoarthritis. AB - The use of glucosamine and chondroitin sulfate for the symptomatic treatment of osteoarthritis has been a subject of controversy for several reasons. First, the medical community in general took offense at the title of Theodosakis' book, The Arthritis Cure. Second, the medical community is becoming divided into "traditional" and "alternative" camps with deep skepticism between them. Third, the whole nutraceutical industry is essentially unregulated, with manufacturers making outrageous claims on products that have never been tested at all, are often of poor quality, and occasionally lacking in any active ingredient. However, for the nutriceuticals evaluated here, there is abundant in vitro, in vivo, animal clinical, and human clinical evidence of both their efficacy and safety. They deserve a prominent place in the armamentarium of nonsurgical treatment of osteoarthritis. PMID- 12730920 TI - The role of debridement: through small portals. AB - Arthroscopic debridement represents one option for treatment of the patient with degenerative arthritis of the knee. Numerous procedures, including joint lavage, removal of loose bodies, partial meniscectomy, and techniques to stimulate cartilage repair may be included under this topic. Understanding surgical indications and clinical results requires differentiation of these individual components where possible. Furthermore, it is important to note that degenerative changes present along a spectrum of involvement. However, despite patient heterogeneity, selective application of conservative debridement, partial meniscectomy and removal of mechanical irritants will result in improved midterm follow-up results in approximately two thirds of patients. Patient expectations are crucial, and the temporizing nature of the procedure must be understood. PMID- 12730921 TI - Cartilage resurfacing: filling defects. AB - Chondral defects with no significant bone involvement can be managed arthroscopically using surface treatments such as debridement and drilling, abrasion arthroplasty, and microfracture. Chondral defects can also be managed arthroscopically using osteochondral autografts (mosaicplasty) or by cartilage cell transplant or periosteal grafts, both of which are performed in open surgery. The arthroscopic surface treatments are best reserved for small defects, but cell transplantation and mosaicplasty have been used for defects up to 3 cm in diameter. Periosteal grafting can be used for large defects affecting an entire condyle, but clinical experience with this procedure is limited and it is still considered investigational. Larger osteochondral defects (uncontained defects greater than 3 cm in diameter and greater than 1 cm in depth) are managed using osteochondral allografts. Realignment osteotomy should be considered in conjunction with any of these techniques in the presence of a coexisting deformity. PMID- 12730922 TI - Instrumentation pitfalls: you just can't go on autopilot! AB - Instrumentation for total knee arthroplasty is crucial for proper positioning and orientation of the components. Instruments, however, must be used properly if the surgeon is to accomplish the goals. The entrance point for intramedullary guides for both the femur and tibia should not be arbitrarily chosen but picked after evaluation of the intersection of the anatomic axes of the respective bones with their articular surfaces. Saw blades must be held on cutting blocks, and the blocks themselves affixed to bones with sufficient pins to assure stability. If an extramedullary tibial alignment guide is chosen, its distal position should be in the center of the ankle plafond, not in the mid-malleolar point. In the lateral plane, it should parallel the shaft of the fibula. Femoral component rotation cannot always be judged from the posterior condylar tangent line. There may be difficulties of deformity with bone and cartilage loss, especially in valgus knees. Surgeons need to be mindful of the epicondylar axis and the midtrochlear of the anteroposterior axis of Whiteside. PMID- 12730923 TI - Total knee arthroplasty in post-traumatic arthrosis of the knee. AB - Total knee arthroplasty is an effective method of treatment for the majority of patients with a prior distal femoral or tibial plateau fracture and end-stage arthritis. There is a higher complication rate and overall poorer outcome when compared with routine primary total knee arthroplasty. In patients for whom the goals of optimal limb and implant alignment are achieved, the results are comparable with routine primary knee arthroplasty. Patients with suboptimal component positioning or residual deformity have a poorer outcome. The technical challenges encountered can require skills, implant systems, and methods usually reserved for complex revision arthroplasty. PMID- 12730924 TI - Total knee arthroplasty rehabilitation protocol: what makes the difference? AB - The goals of any rehabilitation protocol should be to control pain, improve ambulation, maximize range of motion,develop muscle strength, and provide emotional support. Over 85% of total knee arthroplasty (TKA) patients will recover knee function regardless of which rehabilitation protocol is adopted. However, the remaining 15% of patients will have difficulty obtaining proper knee function secondary to significant pain, limited preoperative motion, or the development of arthrofibrosis. This subset will require a special, individualized rehabilitation program that may involve prolonged oral analgesia, continued physical therapy, additional diagnostic studies, and occasionally manipulation. Controlling pain is the mainstay of any treatment plan. The program described herein has been used at the Ranawat Orthopaedic Center over the past 10 years in more than 2,000 TKAs. PMID- 12730925 TI - Orthopaedic crossfire--All patellae should be resurfaced during primary total knee arthroplasty: in the affirmative. AB - Patients with inflammatory arthritis, crystalline deposition disease, or severe patellofemoral arthritis have superior functional results with patellar resurfacing as compared with patellar retention. The only remaining issue is whether there is any advantage to resurfacing or retention in patients with osteoarthritic knees in the absence of severe patellofemoral arthritis. Although the answer is still unclear from available prospective randomized clinical trials, the evidence leans toward routine resurfacing of the patella during primary total knee arthroplasty in osteoarthritic knees too. Evidence exists that the results of patellar retention in osteoarthritic knees deteriorate with longer follow-up study. Long-term follow-up evaluations of available prospective clinical trials will be needed to assess this controversy more definitively. PMID- 12730926 TI - Orthopaedic crossfire--All patellae should be resurfaced during primary total knee arthroplasty: in opposition. AB - After more than 20 years of debate, the decision on whether to resurface the patella in primary total knee arthroplasty (TKA) remains controversial. Laboratory testing suggests that performing a TKA alters knee kinematics. Every study to date has suggested that kinematics are more abnormal when the patella is resurfaced than when it is retained. In addition, patellofemoral contact areas are higher and contact stresses are lower with the native patella compared with the resurfaced patella after TKA. Virtually every clinical study of bilateral knee arthroplasty in which one patella has been resurfaced and the other has not has shown either equivalent results or a preference for the unresurfaced side. Laboratory and clinical data indicate that not resurfacing the patellar is a viable if not preferable option in most TKA patients. PMID- 12730927 TI - Why knees fail. AB - Successful revision knee arthroplasty requires an accurate understanding of the cause of failure. Because a differential diagnosis is necessary, 9 causes of failure are proposed: i) aseptic loosening with or without osteolysis or progression of arthritis in a unicondylar arthroplasty; ii) tibial femoral instability because of collateral ligament instability; iii) patellar complications and malrotation; iv) no diagnosis, the so-called mystery knee; v) structural failure of the implant; vi) sepsis; vii) extensor mechanism rupture; viii) stiffness; and ix) fracture. A revision surgery should correct shortcomings in the original arthroplasty and not simply restore a mechanical situation that has failed. PMID- 12730928 TI - Instability in total knee arthroplasty: loose as a goose. AB - As total knee arthroplasties last longer and are used in younger patients, as well as for a variety of other reasons, greater interest in postoperative tibiofemoral instability has developed. Initial evaluation emphasizing correlation of symptoms and findings together with elucidation of the specifics of the instability are mandatory. Gap inequality, gap asymmetry, and the causes of each are noted. Soft tissue repair or reconstruction alone has not been very successful for such instabilities. Some aspect of revision with component change is generally necessary, and some correction of alignment, gap features, and other issues, with advancement to a higher order of constraint, will generally be necessary. PMID- 12730929 TI - Bone loss in revision total knee arthroplasty: graft options and adjuncts. AB - The treatment of small bone defects in revision total knee arthroplasty should make immediate full weight bearing and full therapy possible, provide long-term stability for the implants, and restore bone stock. These objectives are achieved with bone grafts if the defects are cavitary or small (involving less than one fourth of the cortical rim) segmental defects. These objectives are achieved with porous metal augments if the defects are segmental and involve more than one fourth of the cortical rim. The treatment of bone loss on the femur must allow the re-establishment of the distal and posterior joint lines as well as provide a firm fixation base for the implants. PMID- 12730930 TI - Constraint in total knee arthroplasty: when and what? AB - Tibiofemoral instability is increasingly recognized as a common mode of failure in total knee arthroplasty. Despite this, there are few published guidelines concerning treatment options, component selection, and expected results. In the treatment of the unstable total knee arthroplasty, cruciate-retaining designs can be considered only in the presence of well-balanced ligaments, including the posterior cruciate ligament. More predictable results are obtained with the use of cruciate-substituting components, but they provide no varus-valgus stability and cannot compensate for severe flexion laxity. Varus-valgus constrained designs cannot provide long-term stability in cases of varus-valgus instability or severe flexion laxity. Such cases are most reliably treated with a linked implant. PMID- 12730931 TI - Case challenges in knee surgery: what would you do? PMID- 12730932 TI - Subtrochanteric shortening and derotational osteotomy in primary total hip arthroplasty for patients with severe hip dysplasia: 5-year follow-up. AB - Twenty-one primary hip arthroplasties were performed in Crowe grade 3 or 4 hip dysplasia using a subtrochanteric shortening osteotomy. Average patient age was 48.2 years. Average follow-up was 5.8 years (minimum, 2 years). Femoral fixation was cemented in 10 hips and uncemented in 11 hips. All acetabular components were uncemented, with 33% requiring structural autograft. Ninety-one percent of femoral osteotomies healed without complication. Two osteotomy nonunions required revision. Two acetabular revisions were performed for malposition and polyethylene failure. Three patients experienced postoperative dislocation. One cemented femoral component was revised for loosening. However, no neurologic deficiencies were identified. Harris hip score improved from 32.5 to 73.6. Limp improved in 60% of patients and dependence on assistive walking device improved in 40% of patients. Subtrochanteric shortening osteotomy is a safe and predictable method of restoring the anatomic hip center in high developmental hip dislocation. However, the complication rate in these patients is higher than primary hip arthroplasty for osteoarthritis. PMID- 12730933 TI - Osteonecrosis: what to do, what to do! AB - Despite extensive literature on the subject, the treatment approaches for osteonecrosis of the femoral head (ONFH) remain controversial. The literature presently available to the orthopaedist suggests that i) ONFH can be identified early; ii) there are reasonable treatments for early stages of ONFH; iii) no single treatment is sufficient for all hips; and iv) there is as yet no predictable permanent treatment for early ONFH. The author's recommended approach for the practicing orthopaedist includes the following: i) develop a high degree of clinical suspicion that the disease is present and understand the role of magnetic resonance imaging in making an accurate diagnosis; ii) understand and use an appropriate staging system for each hip diagnosed; iii) be comfortable with several treatment options for early ONFH or choose to refer to appropriate centers; and iv) become comfortable with the use of uncemented THA for end-stage ONFH. The field continues to need and should support better treatment studies, including randomized clinical trials. PMID- 12730934 TI - Acetabular options. AB - Cemented acetabular options have failed anywhere between 10% and 23% over the years and have not improved with increasing cement technology. Cementless technology, however, has basically stopped the problems of prosthesis loosening; however, wear has become a major concern. With present day cement techniques, no radiolucent lines, and a compression-molded polyethylene acetabular component, there is less than a 1% failure rate. The question is, however, whether cementing is technically feasible. Even for experienced surgeons, there is at least a 10% chance of a radiolucent line. Therefore, clinicians should work to develop better bearing surfaces so that cementless technology can be used extensively. PMID- 12730935 TI - Orthopaedic crossfire--Larger femoral heads: a triumph of hope over reason! In the affirmative. AB - The authors' wear studies of total hip arthroplasty cohorts have shown that less polyethylene wear and less deleterious effects of third body debris were found when smaller femoral head sizes were used. The authors' sliding-distance-coupled finite element model findings were corroborated by these clinical wear studies. Thus, with polyethylene on metal bearing surfaces, less wear should occur when smaller head sizes are used. Careful, precise component positioning is important to prevent dislocation. PMID- 12730937 TI - Stability and leg length equality in total hip arthroplasty. AB - Limp is one of the most frustrating persistent symptoms after hip arthroplasty for both the patient and surgeon. There are a multitude of causes of limp; however, leg-length inequality is the leading culprit. Leg-length inequality can lead to anger, litigation, morbidity, and ultimately revision surgery. However, geometrically equal leg lengths may not be an attainable goal in all patients. The stability of the total hip arthroplasty is intimately related to leg length, and equality may need to be sacrificed for stability. Therefore, both the patient and the surgeon should have realistic expectations and an awareness of this problem. PMID- 12730936 TI - Orthopaedic crossfire--Larger femoral heads: a triumph of hope over reason! In opposition. AB - For the past decade, large femoral heads have for the most part been avoided in total hip arthroplasty, especially in younger active patients, because of their association with increased polyethylene wear. However, it has become equally clear that large femoral heads offer a benefit in terms of hip stability and reducing postoperative dislocation. With increasing head size, there is less chance of component-on-component impingement and increased displacement for dislocation. Highly cross-linked polyethylene appears to be relatively insensitive to head size and may offer the surgeon the opportunity to use large femoral heads, especially in patients who are at high risk for dislocation. PMID- 12730938 TI - Controlled femoral fracture: easy in. AB - Revision femoral surgery is a demanding procedure with the potential for severe intraoperative complications, including bone loss and femoral fracture. The extended trochanteric osteotomy is a reproducible and safe technique to remove the femoral components for infection, loosening, component failure, malposition, and dislocation. From 1992 to 1996, 142 consecutive hip revisions were performed with the use of an extended proximal femoral osteotomy. This technique allowed component extraction without fracture in all patients and subsequently allowed for neutral reaming of the femoral canal with placement of the revision stems in proper alignment. There were 2 nonunions of the osteotomized fragments at an average postoperative follow-up period of 2.6 years. Additional complications included 4 fractures of the osteotomized fragment and one malunion. We have found that use of the osteotomy is an efficient, safe and reliable technique in revision hip arthroplasty. PMID- 12730940 TI - Orthopaedic crossfire--Stem modularity is unnecessary in revision total hip arthroplasty: in the affirmative. AB - Numerous options are available for femoral stem fixation in revision total hip arthroplasty, including cemented stems, proximally-coated cementless stems, extensively coated cementless stems, impaction grafting cemented stems, and modular cementless stems. In terms of versatility, ease of insertion, use with an extended trochanteric osteotomy, and clinical results, extensively coated cementless stems remain the gold standard. Modular cementless stems have not matched the clinical results of extensively coated cementless stems. In addition, modular stems introduce increased complexity, cost, and potential complications and are therefore, rarely, if ever, necessary in revision total hip arthroplasty. PMID- 12730941 TI - Orthopaedic crossfire--Stem modularity is unnecessary in revision total hip arthroplasty: in opposition. AB - If distal fixation is a goal in revision hip surgery, then modularity may not be necessary. If, however, proximal fixation is desired, both proximal and distal fit and fill are necessary to achieve initial stability. This can only be done in the revision situation by using a modular stem. Long-term follow up of 320 revision cases treated using a proximally modular proximal ingrowth stem shows a re-revision rate for late aseptic loosening of 0.93% at a mean time of 7 years, indicating that such a stem is a reasonable option. PMID- 12730939 TI - A modular distal fixation option for proximal bone loss in revision total hip arthroplasty: a 2- to 6-year follow-up study. AB - Proximal femoral bone deficiency in the failed total hip arthroplasty (THA) is addressed with difficulty using proximally fixed implants. The Link MP reconstruction hip stem is proximally modular but utilizes distal fixation. This series consists of 143 patients from 3 clinical sites. Of the patients, 81 were women. The average follow-up time was 40 months, and the average patient age was 67 years. The preoperative diagnoses included 108 cases of aseptic loosening and 14 periprosthetic fractures. Clinical follow-up of all patients included radiographic assessment of the hip. This study found a 97.2% survival rate. The average subsidence was 2.1 mm. The average postoperative Harris hip score was 92. Complications included 7 wound infections and 4 deep venous thromboses. We found that the Link MP hip stem allows successful revision THA reconstruction of the proximally compromised femur. Good to excellent functional restoration and pain relief is achievable. PMID- 12730942 TI - Onlay cortical strut grafting in revision arthroplasty of the hip. AB - Cortical strut allografts are used to restore uncontained noncircumferential femoral defects, to bypass stress risers, to stabilize proximal femoral allograft host junctions, and to fix periprosthetic fractures. These struts can be made from hemicylinders of diaphyseal bone or from fibulae. They are fixed to host bone by circumferential wires. If autograft bone is available, it is placed between the strut and host, particularly at the ends of the strut. A radiographic study of 52 cortical strut allografts with an average follow-up of 4.8 years revealed 2 nonunions and 2 progressive resorptions. All other struts united with minimal resorption. PMID- 12730943 TI - Periprosthetic fractures associated with osteolysis: a problem on the rise. AB - Periprosthetic fractures caused by osteolysis can occur in the pelvis, peritrochanteric area, or femoral diaphysis after hip arthroplasty. Most, but not all, hips are ultimately treated surgically to manage the osteolysis or to stabilize the fracture. Some stable, minimally displaced fractures of the pelvis and greater trochanter can be allowed to heal before revision is considered, which simplifies the operation. Unstable fractures, especially of the femoral diaphysis, are usually treated with early repeat surgery following the established principles of periprosthetic fracture management. PMID- 12730944 TI - Management of patients with acetabular socket wear and pelvic osteolysis. AB - Polyethylene wear and pelvic osteolysis are the most common late complications associated with stable cementless total hip implants. This manuscript describes the diagnostic strategies and treatment algorithm used at the senior author's (C.A.E) institution for patients with wear and pelvic osteolysis. This evolving management strategy is based on our experiences and ongoing research. We discuss patient selection, the evaluation of acetabular liner wear, the diagnosis of pelvic osteolysis, the timing of revision, and treatment strategies. According to this algorithm, we revise asymptomatic hips with pending or complete wear-through of the acetabular liner. We also recommend revision for most symptomatic patients with pelvic osteolysis and for patients with large pelvic osteolytic bone defects or a documented increase in osteolytic lesion size in an area of the cementless acetabular component in which a load transfer between the implant and the surrounding bone is likely to occur. PMID- 12730945 TI - Cementless acetabular components: the gold standard for socket revision. AB - Hemispherical ingrowth sockets are used for acetabular revision in over 90% of cases. Multiple experiences now show the success of this technique in both consecutive series and specific settings. In addition to understanding the technique required to implant these components successfully, the surgeon must be aware of the settings in which the construct is not likely to work and plan for alternative techniques when needed. PMID- 12730946 TI - Management of massive acetabular defects in revision total hip arthroplasty. AB - Twenty-four cases of massive periacetabular bone loss (Paprosky Type 3B) were reconstructed using custom triflanged acetabular components (CTAC). Fixation is obtained by the creation of a triflanged component that is anchored to the ilium, ishium, and pubis with multiple fixation screws. Acetabular defects are grafted with large amounts of cancellous allograft. Modular polyethylene liners are then locked into the CTAC. Twenty-one of 24 cases (87.5%) were considered clinically successful at short-term follow-up (average, 48 months; maximum, 78 months), with stable fixation and reconstruction of periacetabular bone. Three failures occurred from loss of fixation in patients with a preoperative dissociation of the hemipelvis and severe osteopenia. Presently, these devices should be used with caution in cases with a preoperative dissociation of the hemipelvis, unless additional column plating is performed. PMID- 12730948 TI - Diffusion tensor imaging in fixed brain tissue at 7.0 T. AB - The purpose of this work is to assess the feasibility of performing quantitative in vitro brain tissue diffusion tensor imaging (DTI) measurements and to examine their comparability to in vivo measurements. DTI of fixed tissue at high field strength is potentially a very valuable investigative tool as very high spatial resolution can be achieved. DTI was applied to human and mouse brain fixed tissue samples as well as in vivo measurements of the mouse brain. T(1) and T(2) relaxography of the fixed tissue samples was also performed to provide further characterization of the tissue. All experiments were performed at 7 T. The fractional anisotropy (FA) of the human fixed brain tissue samples is found to be higher in the corpus callosum than in the occipital white matter region, consistent with in vivo measurements reported in the literature. Our FA measurements of the corpus callosum of a mouse brain are also found to be the same both in vitro and in vivo. This preliminary work supports the use of DTI in both fixed human and fixed animal brain tissue as a valid investigative tool. With the increased availability of brain banks in different brain disorders, DTI in fixed tissue may prove to be a very useful method for the study of white matter abnormalities. PMID- 12730949 TI - Manganese ions as intracellular contrast agents: proton relaxation and calcium interactions in rat myocardium. AB - Paramagnetic manganese (Mn) ions (Mn(2+)) are taken up into cardiomyocytes where they are retained for hours. Mn content and relaxation parameters, T(1) and T(2), were measured in right plus left ventricular myocardium excised from isolated perfused rat hearts. In the experiments 5 min wash-in of MnCl(2) were followed by 15 min wash-out to remove extracellular (ec) Mn(2+) MnCl(2), 25 and 100 micro M, elevated tissue Mn content to six and 12 times the level of control (0 micro M MnCl(2)). Variations in perfusate calcium (Ca(2+)) during wash-in of MnCl(2) and experiments including nifedipine showed that myocardial slow Ca(2+) channels are the main pathway for Mn(2+) uptake and that Mn(2+) acts as a pure Ca(2+) competitor and a preferred substrate for slow Ca(2+) channel entry. Inversion recovery analysis at 20 MHz revealed two components for longitudinal relaxation: a short T(1 - 1) and a longer T(1 - 2). Approximate values for control and Mn treated hearts were in the range 600-125 ms for T(1 - 1) and 2200-750 ms for T(1 2). The population fractions were about 59 and 41% for the short and the long component, respectively. The intracellular (ic) R(1 - 1) and R(2 - 1) correlated best with tissue Mn content. Applying two-site exchange analyses on the obtained T(1) data yielded results in parallel to, but also differing from, results reported with an ec contrast agent. The calculated lifetime of ic water (tau(ic)) of about 10 s is compatible with a slow water exchange in the present excised cardiac tissue. The longitudinal relaxivity of Mn ions in ic water [60 (s mM)( 1)] was about one order of magnitude higher than that of MnCl(2) in water in vitro [6.9 (s mM)(-1)], indicating that ic Mn-protein binding is an important potentiating factor in relaxation enhancement. PMID- 12730947 TI - Evaluation of (E)-2'-deoxy-2'-(fluoromethylene)cytidine on the 9L rat brain tumor model using MRI. AB - (E)-2'-deoxy-2'-(fluoromethylene)cytidine (FMdC), was evaluated as a potential treatment for malignant gliomas using the rat 9L brain tumor model. FMdC was shown to be an effective inhibitor of cell proliferation in cultured 9L cells with an EC(50) of 40 ng/ml. In vitro studies also revealed that this compound significantly inhibited incorporation of [(3)H]thymidine in 9L cells. In vivo therapeutic efficacy of FMdC was evaluated in rats harboring intracerebral 9L tumors which were treated daily with 15 mg/kg, i.p. Treatment response was quantified from changes in tumor growth rates as assessed from sequential magnetic resonance imaging (MRI) tumor volume measurements. In vivo tumor cell kill in individual animals was calculated by fitting tumor volume data with an iterative computer routine. It was estimated that therapeutically responsive rats treated with FMdC daily produced a >/= 0.1 log kill per therapeutic dose which resulted in a significant reduction in tumor growth rate. In addition, localized (1)H-MRS of intracerebral 9L tumors revealed changes in metabolite levels which correlated with therapeutic response. These results provide evidence supporting the use of FMdC in clinical trials for the treatment of malignant gliomas and reveals that MR can play an important role in the pre-clinical evaluation of novel compounds using orthotopic tumor models. PMID- 12730950 TI - Leakage of metabolites from tissue biopsies can result in large errors in quantitation by MRS. AB - The leakage of metabolites from frozen and thawed tissue biopsies was measured semi-quantitatively by high-field (8.5 T) proton MRS. Human prostate and rat brain tissue specimens, frozen within 1 min of collection, lost significant and variable amounts of diagnostic metabolites immediately upon thawing. For prostate tissue 30-50% of initial total choline compounds, total creatines and citrate were detected in the collection buffer immediately after thawing. The widely used protocol for MR assessment of tissue biopsies, which involves washing of thawed tissue samples in fresh buffer, results in loss of large and unpredictable amounts of possibly diagnostic metabolites prior to MRS. This reduces the reproducibility of MR analysis of tissue biopsies and compromises the reliable identification of MR spectral patterns diagnostic of tissue pathology. The problem can be avoided by minimizing the volume of storage buffer, omitting tissue washing and performing MRS measurements on the tissue immersed in the original storage buffer. PMID- 12730951 TI - Detecting early response to cyclophosphamide treatment of RIF-1 tumors using selective multiple quantum spectroscopy (SelMQC) and dynamic contrast enhanced imaging. AB - The purpose of this study was to develop a reliable, noninvasive method for early detection of tumor response to therapy that would facilitate optimization of treatment regimens to the needs of the individual patient. In the present study, the effects of cyclophosphamide (Cp, a widely used alkylating agent) were monitored in a murine radiation induced fibrosarcoma (RIF-1) using in vivo (1)H NMR spectroscopy and imaging to evaluate the potential of these techniques towards early detection of treatment response. Steady-state lactate levels and Gd DTPA uptake kinetics were measured using selective multiple quantum coherence (Sel-MQC) transfer spectroscopy and dynamic contrast enhanced imaging, respectively in RIF-1 tumors before, 24 and 72 h after 300 mg/kg of Cp administration. High-resolution (1)H NMR spectra of perchloric acid extracts of the tumor were correlated with lactate and glucose concentrations determined enzymatically. In vivo NMR experiments showed a decrease in steady-state lactate to water ratios (5.4 +/- 1.6 to 0.6 +/- 0.5, p < 0.05) and an increase in Gd-DTPA uptake kinetics following treatment response. The data indicate that decreases in lactate result from decreased glycolytic metabolism and an increase in tumor perfusion/permeability. Perchloric acid extracts confirmed the lower lactate levels seen in vivo in treated tumors and also indicated a higher glycerophosphocholine/phosphocholine (GPC/PC) integrated intensity ratio (1.39 +/ 0.09 vs 0.97 +/- 0.04, p < 0.01), indicative of increased membrane degradation following Cp treatment. Steady-state lactate levels provide metabolic information that correlates with changes in tumor physiology measured by Gd-DTPA uptake kinetics with high spatial and temporal resolution. Both of these parameters may be useful for monitoring early tumor response to therapy. PMID- 12730952 TI - 14-3-3gamma is upregulated by in vitro ischemia and binds to protein kinase Raf in primary cultures of astrocytes. AB - The 14-3-3 protein family comprises critical regulatory molecules involved in signaling during cell division, proliferation, and apoptosis. Despite extensive study, the functions of the 14-3-3 proteins in brain remain unclear. 14-3-3gamma, a subtype of the 14-3-3 family of proteins, was thought to be brain- and neuron specific. Using RNA arbitrarily primed PCR, we identified an upregulated cDNA fragment of the 14-3-3gamma gene in primary cultures of astrocytes. Using Northern blot analysis, we confirmed this fragment was brain-specific. In cultures of astrocytes, 14-3-3gamma genes and proteins were differentially expressed at different ages and the proteins were distributed only in the cytoplasm. These results indicated that 14-3-3gamma was not neuron-specific but also expressed in astrocytes. The function of this protein in brain is unclear. Northern and Western blot analyses demonstrated that 14-3-3gamma mRNA and protein were upregulated in cultured astrocytes in an anaerobic chamber-induced ischemia model. The induction of 14-3-3gamma proteins was neither suppressed by an MAP kinase inhibitor (U0126) nor a PI-3 kinase inhibitor (LY294002). These data indicated that induction of 14-3-3gamma might not involve PI-3 and MAP kinase dependent pathways. Using coimmunoprecipitation, we demonstrated that endogenous 14-3-3gamma bound to c-Raf-1 and p-Raf 259. As Raf is one of the critical serine/threonine kinases controlling cell growth, differentiation, and death, the binding of 14-3-3gamma to Raf indicates the critical role of this protein in ischemia-induced apoptosis and the changes in signal transduction in astrocytes in culture. PMID- 12730953 TI - Chlorotoxin-sensitive Ca2+-activated Cl- channel in type R2 reactive astrocytes from adult rat brain. AB - Astrocytes express four types of Cl(-) or anion channels, but Ca(2+)-activated Cl(-) (Cl(Ca)) channels have not been described. We studied Cl(-) channels in a morphologically distinct subpopulation ( approximately 5% of cells) of small (10 12 micro m, 11.8 +/- 0.6 pF), phase-dark, GFAP-positive native reactive astrocytes (NRAs) freshly isolated from injured adult rat brains. Their resting potential, -57.1 +/- 4.0 mV, polarized to -72.7 +/- 4.5 mV with BAPTA-AM, an intracellular Ca(2+) chelator, and depolarized to -30.7 +/- 6.1 mV with thapsigargin, which mobilizes Ca(2+) from intracellular stores. With nystatin perforated patch clamp, thapsigargin activated a current that reversed near the Cl(-) reversal potential, which was blocked by Cl(-) channel blockers, 5-nitro-2 (3-phenylpropylamino)-benzoate (NPPB) and Zn(2+), by I(-) (10 mM), and by chlorotoxin (EC(50) = 47 nM). With conventional whole-cell clamp, NPPB- and Zn(2+)-sensitive currents became larger with increasing [Ca(2+)](i) (10, 150, 300 nM). Single-channel recordings of inside-out patches confirmed Ca(2+) sensitivity of the channel and showed open-state conductances of 40, 80, 130, and 180 pS, and outside-out patches confirmed sensitivity to chlorotoxin. In primary culture, small phase-dark NRAs developed into small GFAP-positive bipolar cells with chlorotoxin-sensitive Cl(Ca) channels. Imaging with biotinylated chlorotoxin confirmed the presence of label in GFAP-positive cells from regions of brain injury, but not from uninjured brain. Chlorotoxin-tagged cells isolated by flow cytometry and cultured up to two passages exhibit positive labeling for GFAP and vimentin, but not for prolyl 4-hydroxylase (fibroblast), A2B5 (O2A progenitor), or OX-42 (microglia). Expression of a novel chlorotoxin-sensitive Cl(Ca) channel in a morphologically distinct subpopulation of NRAs distinguishes these cells as a new subtype of reactive astrocyte. PMID- 12730954 TI - Early onset of degenerative changes at nodes of Ranvier in alpha-motor axons of Cntf null (-/-) mutant mice. AB - The nodes of Ranvier are sites of specific interaction between Schwann cells and axons. Besides their crucial role in transmission of action potentials, the nodes of Ranvier and in particular the paranodal axon-Schwann cell networks (ASNs) are thought to function as local centers in large motor axons for removal, degradation, and disposal of organelles. In order to test whether ciliary neurotrophic factor (CNTF), which is present at high levels in the Schwann cell cytoplasm, is involved in the maintenance of these structures, we have examined lumbar ventral root nerve fibers of alpha-motor neurons by electron microscopy in 3- and 9-month-old Cntf null ((-/-)) mutant mice. Nerve fibers and nodes of Ranvier in 3-month-old Cntf(-/-) mutants appeared morphologically normal, except that ASNs were more voluminous in the mutants than in wild-type control animals at this age. In 9-month-old Cntf(-/-) animals, morphological changes, such as reduction in nerve fiber and axon diameter, myelin sheath disruption, and loss of ASNs at nodes of Ranvier, were observed. These findings suggest that endogenous CNTF, in addition to its role in promoting motor neuron survival and regeneration, is needed for long-term maintenance of alpha-motor nerve fibers. The premature loss of paranodal ASNs in animals lacking CNTF, which seems to be a defect related to a disturbed interaction in the nodal region between the axon and its myelinating Schwann cells, could impede the maintenance of a normal milieu in the motor axon, preceding more general neuronal damage. PMID- 12730956 TI - Thyroid hormone modulates the extracellular matrix organization and expression in cerebellar astrocyte: effects on astrocyte adhesion. AB - The effects of thyroid hormone (T(3)) on extracellular matrix (ECM) expression and organization in cerebellar astrocytes were studied. Control astrocytes exhibit laminin immunostaining distributed in a punctate configuration and fibronectin concentrated in focal points at the cell surface. These cells attach to the substratum by membrane points, as shown by scanning microscopy, possibly by focal points stained to fibronectin. In contrast, after T(3) treatment, laminin assumes a fibrillary pattern and fibronectin becomes organized in filaments homogeneously distributed on the cell surface; the cells acquire a very flat and spread morphology. T(3) treatment also modulates astrocyte adhesion. In addition, increased expression of both laminin and fibronectin was detected by Western blot. These alterations in fibronectin and/or laminin production and organization may be involved in the flat and spread morphology and in altered adhesion. We observed that fibroblast growth factor-2 (FGF(2)) added to cultures had similar effects to those described to T(3). Neutralizing antibodies against FGF(2) reversed T(3) effects on fibronectin and laminin distribution. We also observed that cerebellar neurons co-cultured on T(3)-treated astrocytes had an increase in the number of cells and presented longer neurites. Thus, we propose a novel mechanism of the effect of thyroid hormone on cerebellar development mediated by astrocytes: T(3) may induce astrocyte secretion of growth factors, mainly FGF(2), that autocrinally stimulate astrocyte proliferation, reorganization in ECM proteins, and alterations in cell spreading and adhesion. These effects may indirectly influence neuronal development. PMID- 12730955 TI - Malignant peripheral nerve sheath tumor cell invasion is facilitated by Src and aberrant CD44 expression. AB - Malignant peripheral nerve sheath tumors (MPNSTs) are aggressive malignancies that arise within peripheral nerves. These tumors occur with increased incidence in patients with neurofibromatosis type 1 (NF1), exhibiting increased Ras activity due to loss of the NF1 gene product, neurofibromin, and abnormal expression of the epidermal growth factor receptor (EGFR). We previously found that MPNSTs express increased levels of the CD44 family of transmembrane glycoproteins that have been implicated in tumor cell invasion and metastasis. In two MPNST cell lines, we have found that elevated CD44 expression and cell invasion are dependent on Src kinase activity but are independent of mitogen activated protein kinases (MAPK) kinase (MEK) activity. In contrast, inhibition of Src kinase activity has no influence on MPNST cell proliferation. Reduction of CD44 levels, using antisense oligonucleotides, results in reduced MPNST cell invasion in vitro, suggesting that Src contributes in part to MPNST cell invasion by increasing CD44 levels. At least some of this increased CD44 expression results from elevated EGFR levels through a Src-dependent mechanism, consistent with the notion that EGFR promotes constitutive Src activation in MPNSTs. These data indicate that Src and CD44 are putative targets for the treatment of MPNST invasion and metastasis. PMID- 12730958 TI - Human malignant glioma cells express semaphorins and their receptors, neuropilins and plexins. AB - Semaphorins comprise a family of molecules implicated in the guiding of growing axons and neuronal progenitor cells. Further, semaphorins have been suggested to play a role in cancer metastasis. Neuropilins 1 and 2 are cell surface receptors for soluble class 3 semaphorins. Plexins are direct receptors for membrane-bound semaphorins and, by binding to neuropilins, coreceptors necessary for class 3 semaphorin signaling. We here report that human malignant glioma cell lines express neuropilins 1 and 2 mRNA and protein, as well as either plexin A1, A2, or B1. Further, all glioma cell lines express SEMA3A and SEMA3C and exhibit SEMA3A binding sites. Exogenous SEMA3A expressed in 293 or U87MG cells has no collapsing or chemorepulsive activities on glioma cells as determined by F-actin staining and collagen coculture assays. In summary, human glioma cells express class 3 semaphorins and receptors for soluble and membrane-bound semaphorins, suggesting a possible role of the semaphorin/neuropilin system in the interactions of human malignant glioma with the host's central nervous and immune systems. PMID- 12730957 TI - Targeting human 8-oxoguanine glycosylase to mitochondria of oligodendrocytes protects against menadione-induced oxidative stress. AB - Within the central nervous system (CNS), there is a differential susceptibility among cell types to certain pathological conditions believed to involve oxidative stress. Oligodendrocytes are extremely sensitive to oxidative stress, which correlates with a decreased ability to repair damage in mitochondrial DNA (mtDNA), as we have shown previously. To determine whether there is a causal relationship, studies were carried out to correct the deficit in repair of the oxidative damage in mtDNA in cultured oligodendrocytes. A vector containing a mitochondrial transport sequence (MTS) upstream of the sequence for human 8 oxoguanine-DNA glycosylase (OGG) was transfected into the cells. The efficiency of transfection and the localization of recombinant protein were determined by fluorescence microscopy and by Western blot analysis. Subsequent mtDNA repair studies, employing 100 micro M menadione to produce reactive oxygen species, showed a significant enhancement in repair of oxidative lesions in mtDNA of MTS OGG transfected oligodendrocytes compared with cells transfected with vector only. Experiments were also conducted to determine the effect of changing mtDNA repair capacity on menadione-induced apoptosis in oligodendrocytes. These experiments show that targeting the OGG repair enzyme to mitochondria reduces the release of cytochrome c from the intermitochondrial space and the activation of caspase 9 in oligodendrocytes after exposure to menadione. Therefore, targeting of DNA repair enzymes to mitochondria appears to be a viable approach for the protection of cells against some of the deleterious effects of oxidative stress. PMID- 12730959 TI - Effect of endothelin-1 on astrocytic protein content. AB - The astrocytic endothelin (ET) receptors, ET(A) and ET(B), modulate calcium signaling and the astrocytic gap junctional network. The nonselective ET receptor ligand ET-1 inhibits gap junction permeability, an effect that can be blocked by tolbutamide. This mechanism may play a role in pathophysiological conditions such as ischemic stroke, characterized by elevated tissue ET-1 levels and hypertrophic appearing reactive astrocytes. Therefore, the effect of ET-1 on cellular protein content was investigated in confluent once-passaged rat astrocyte cultures under serum-free conditions, by the Lowry method. Gap junction permeability was determined by the dye transfer technique. ET-1 prevented the decrease in astrocytic protein content observed in controls. The effect of ET-1 on cellular protein content was most pronounced in cultures seeded at high density, but it was attenuated in ET(B)-deficient (sl/sl) astrocytes. This effect could be blocked by the nonselective ET antagonist LU 302872 (10 micro M), as well as by the protein synthesis inhibitor cycloheximide (10 micro M). This increase in astrocytic protein content was inhibited by the ATP-sensitive K(+) channel blocker tolbutamide, which also antagonized the ET-1-induced reduction of gap junction permeability and reversed the morphological changes observed in astrocytes upon ET-1 treatment. Cytosine arabinoside (10 micro M), a DNA synthesis blocker, inhibited the ET-1-induced BrdU uptake without affecting the ET-1-induced increase in astrocytic protein content. To conclude, ET-1 induces an increase in astrocytic protein content as well as changes in astrocyte morphology in vitro. This hypertrophic response involves uncoupling of the astrocytic gap junctional network and is not dependent on DNA synthesis. PMID- 12730961 TI - Differential effects of apotransferrin on two populations of oligodendroglial cells. AB - In the central nervous system (CNS), apotransferrin (aTf) is produced by oligodendroglial cells (OLGcs), and aTf is essential for cell survival. We previously demonstrated that a single intracranial injection of aTf in 3-day-old rats accelerates differentiation of OLGc and that aTf acts at early stages of development on certain populations of OLGcs, promoting accelerated maturation, with no effect on late markers of cell differentiation. The objective of the present study was to analyze OLGc maturation at two different stages of rat development, 4 and 10 days of age, in OLGcs isolated from the brain after intracranial injection of aTf at 3 days of age, and to explore the in vitro effect of aTf added to cultures of OLGc isolated from aTf-injected and control brains. The maturational cell stages were identified by immunocytochemistry with different OLGc markers and by analysis of their morphological complexity. The OLGcs isolated from 4- and 10-day-old animals intracranially injected with aTf were more differentiated than control cells. Treatment with aTf of the cultures of OLGcs that were isolated from 4-day-old saline-injected control animals induced their differentiation, while a similar treatment of the cultures of OLGcs that were isolated from 10-day-old animals did not induce further maturation of the cells. The results presented in the present report demonstrate that the in vivo effects of aTf on OLGc maturation can be reproduced in cultures and that the effects of aTf occur early in development during a narrow, transient "temporal window" within which OLGcs are sensitive to its action. PMID- 12730962 TI - Expression of complement messenger RNAs and proteins by human oligodendroglial cells. AB - Neurons, astrocytes, microglia, and endothelial cells are capable of synthesizing most, if not all, of the complement proteins. Little is known, however, about the capacity of oligodendroglial cells to generate complement components. This study evaluated expression of complement mRNAs and their protein products by human oligodendrocytes. Cells were isolated and cultured from white matter of seven adult cases that had undergone surgical temporal lobe resection for epilepsy. Oligodendroglial cultures were characterized by the expression of such cell type specific mRNAs as myelin proteolipid protein (PLP), oligodendrocyte-specific protein (OSP), and 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) and were further characterized by immunostaining for such differentiation markers as myelin basic protein (MBP), PLP, CNPase, and O4. RT-PCR analysis showed that the oligodendroglial cells expressed detectable levels of complement mRNAs for the C1q B-chain, C1r, C1s, C2, C3, C4, C5, C6, C7, C8 gamma subunit, and C9. Immunostaining was positive for C1q, C1s, C2, C3, C4, C5, C6, C7, C8, and C9. Double immunostaining for the oligodendrocyte marker O4 and the complement protein C3 demonstrated that all O4-positive cells were also positive for C3, indicating constitutive C3 expression. These results indicate that oligodendroglial cells may be a source of complement proteins in human brain and thus could contribute to the pathogenesis of several neurodegenerative and inflammatory diseases of the CNS, such as Alzheimer's disease, multiple sclerosis, and progressive supranuclear palsy, where complement-activated oligodendrocytes are abundant. PMID- 12730960 TI - ADAM-10 and ADAM-17 in the inflamed human CNS. AB - Inflammatory demyelinating disorders of the CNS, such as multiple sclerosis (MS), are mediated, at least in part, by various cytokines and proteases. In the present study, we investigated the expression of A disintegrin and metalloproteinase (ADAM)-17, an important sheddase for various proteins, including tumor necrosis factor-alpha (TNF-alpha), and the p75- and p55-TNF receptors, as well as ADAM-10, a protease implicated in myelin degradation, in post mortem CNS tissue samples from patients with MS, and normal brain tissue (as control) by immunohistochemistry. ADAM-10 was found to be expressed by astrocytes in all MS and control sections studied; however, in some MS sections, perivascular macrophages were determined as an additional cellular source as well. ADAM-17 could be observed exclusively in acute and chronic active MS plaques and localized to invading T lymphocytes. The staining pattern of ADAM-17 in MS plaques was mirrored in distribution and extent by the pattern obtained with an antibody against the p75-TNF-receptor (TNFR-2), whereas TNF-alpha was found to be expressed primarily by perivascular macrophages. In studying cerebrospinal fluid (CSF) samples from MS patients, we were able to detect increased protein levels of ADAM-17 as compared with noninflammatory controls. In addition, increased levels of soluble TNFR-2 could be measured, suggestive of an active shedding process mediated by ADAM-17. The stimulation of peripheral blood mononuclear cells (PBMC) obtained from MS patients and healthy individuals corroborated these findings by revealing expression of ADAM-17 by T lymphocytes and ADAM-10 by macrophages in vitro. Our results indicate that ADAM-10 is expressed constitutively by astrocytes in the normal and inflamed human CNS. In contrast, under inflammatory conditions, ADAM-10, expressed by perivascular macrophages, and ADAM-17, expressed by invading T cells, may actively contribute to the pathogenesis of inflammatory disorders of the CNS. PMID- 12730963 TI - Sequential loss of myelin proteins during Wallerian degeneration in the rat spinal cord. AB - Axotomy of nerve fibers leads to the subsequent degeneration of their distal part, a process termed Wallerian degeneration (WD). While WD in the peripheral nervous system is usually followed by regeneration of the lesioned axons, central nervous system (CNS) neurons are generally unable to regrow. In this study, we investigated the process of WD in the dorsal columns of the rat spinal cord rostral to a mid-thoracic lesion. We confirm earlier studies describing a very delayed microglial and an early and sustained astroglial reaction finally leading to scar formation. Interestingly, we found a differential time course in the loss of myelin proteins depending on their location. Proteins situated on the periaxonal myelin membrane such as myelin associated glycoprotein disappeared early, within a few days after lesion, concomitantly with cytoskeletal axonal proteins, whereas compact myelin and outer myelin membrane proteins such as MBP and Nogo-A remained for long intervals in the degenerating tracts. Two distinct mechanisms are probably responsible for this difference: processes of protein destruction emanating from and initially probably located in the axon act on a time scale of 1-3 days. In contrast, the bulk of myelin destruction is due to phagocytosis known to be slow, prolonged, and inefficient in the CNS. These results may also have implications for future intervention strategies aiming at enhancing CNS regeneration. PMID- 12730967 TI - Gold goes nano--from small clusters to low-dimensional assemblies. PMID- 12730968 TI - Supramolecular crown ether adducts in the gas phase: from molecular recognition of amines to the covalent coupling of host/guest molecules. PMID- 12730964 TI - Oncostatin M enhances the expression of prostaglandin E2 and cyclooxygenase-2 in astrocytes: synergy with interleukin-1beta, tumor necrosis factor-alpha, and bacterial lipopolysaccharide. AB - Oncostatin M (OSM), a cytokine of the interleukin-6 family, is expressed in rheumatoid arthritis, multiple sclerosis, multiple myeloma, and other inflammatory and neoplastic conditions. Prostaglandin E(2) (PGE(2)), an eicosanoid also associated with inflammation and cancer, has recently been shown to induce OSM expression. We report here that OSM in turn induces PGE(2) production by astrocytes and astroglioma cells. More importantly, in combination with the inflammatory mediators IL-1beta, tumor necrosis factor-alpha, and lipopolysaccharide, OSM exhibits a striking synergy, resulting in up to 50-fold higher PGE(2) production by astrocytes, astroglioma, and neuroblastoma cell lines. Enhanced PGE(2) production by OSM and IL-1beta treatment is explained by their effect on cyclooxygenase-2 (COX-2), an enzyme that catalyzes the committed step in PGE(2) synthesis. Of the enzymes involved in PGE(2) biosynthesis, only COX-2 mRNA and protein levels are synergistically amplified by OSM and IL-1beta. Nuclear run-on assays demonstrate that OSM and IL-1beta synergistically upregulate transcription of the COX-2 gene, and the mRNA stability assay indicates that COX-2 mRNA is posttranscriptionally stabilized by OSM and IL 1beta. To effect synergy on the PGE(2) level, OSM signals in part through its gp130/OSMRbeta receptor, since neutralizing antibodies against gp130 and OSMRbeta, but not LIFRbeta, decrease PGE(2) production in response to OSM plus IL 1beta. SB202190 and U0126, inhibitors of p38 MAPK and ERK1/2 activation, respectively, inhibit IL-1beta and OSM upregulation of COX-2 and PGE(2), indicating that these MAPK cascades are utilized by both stimuli. This mechanism of PGE(2) amplification may be active in brain pathologies where both OSM and IL 1beta are present, such as glioblastomas and multiple sclerosis. PMID- 12730969 TI - Recent developments in olefin cross-metathesis. AB - Among the many types of transition-metal-catalyzed C-C bond-forming reactions, olefin metathesis has come to the fore in recent years owing to the wide range of transformations that are possible with commercially available and easily handled catalysts. Consequently, olefin metathesis is now widely considered as one of the most powerful synthetic tools in organic chemistry. Until recently the intermolecular variant of this reaction, cross-metathesis, had been neglected despite its potential. With the evolution of new catalysts, the selectivity, efficiency, and functional-group compatibility of this reaction have improved to a level that was unimaginable just a few years ago. These advances, together with a better understanding of the mechanism and catalyst-substrate interactions, have brought us to a stage where more and more researchers are employing cross metathesis reactions in multistep procedures and in the synthesis of natural products. The recent inclusion of alkynes and hindered bicyclic olefins as viable substrates for bimolecular metathesis coupling, the discovery of enantioselective cross-metathesis and cross-metathesis in water, and the successful marriage of metathesis and solid-phase organic synthesis has further widened the scope of this versatile reaction. PMID- 12730970 TI - Kinetics of a three-step reaction observed at the single-molecule level. PMID- 12730972 TI - Covalent connection of two individual polymer chains on a surface: an elementary step towards molecular nanoconstructions. PMID- 12730971 TI - Stabilization of beta-SiB3 from liquid Ga: a boron-rich binary semiconductor resistant to high-temperature air oxidation. PMID- 12730973 TI - The molecular structure of [Os3Rh4(mu3-eta1:eta1:eta1-C6H5CH3)(CO)13]: a face capping bonding mode for arenes in organometallic clusters. PMID- 12730974 TI - NCHU-3: a crystalline inorganic-organic hybrid molecular sieve with extra-large cages. PMID- 12730975 TI - Synthesis of a self-assembled molecular capsule that traps pyridine molecules by a combination of hydrogen bonding and copper(II) coordination. PMID- 12730976 TI - Templated synthesis of inorganic hollow spheres with a tunable cavity size onto core-shell gel particles. PMID- 12730977 TI - Spontaneous formation of ordered 3D superlattices of nanocrystals from polydisperse colloidal solutions. PMID- 12730978 TI - Novel azine reactivity: facile N-N bond cleavage, C-H activation, and N-N coupling mediated by RhI. PMID- 12730979 TI - Isolation of a uranyl [UO2]+ species: crystallographic comparison of the dioxouranium(V) and (VI) compounds [UO2(OPPh3)4](OTf)n (n=1, 2). PMID- 12730980 TI - "Walking" of the C-C pi bond over long distances in Pd-catalyzed reactions of 2,3 allenoic acids with omega-1-alkenyl halides. PMID- 12730981 TI - A classical example of a disappearing polymorph and the shortest intermolecular H...H separation ever found in an organic crystal structure. PMID- 12730982 TI - Cocrystallization with acetylene: molecular complexes with acetone and dimethyl sulfoxide. PMID- 12730983 TI - Searching for a polymorph: second crystal form of 6-amino-2-phenylsulfonylimino 1,2-dihydropyridine. PMID- 12730985 TI - [Ga16(PtBu2)10]: a gallium phosphide sheathed core of four naked Ga atoms? PMID- 12730984 TI - Peptide-PNA conjugates: targeted transport of antisense therapeutics into tumors. PMID- 12730986 TI - Reactivity of osmium tetraoxide towards nitrogen heterocycles: implications for the molecular recognition of DNA mismatch. PMID- 12730988 TI - Diminished auditory mismatch fields in dyslexic adults. AB - Electroencephalographic studies have demonstrated smaller auditory responses to infrequent deviances of speech and nonspeech sounds in dyslexic than normal reading subjects. We used a whole-scalp neuromagnetometer to study selectively reactivity of the auditory cortices to sound deviances in 8 dyslexic and 11 normal-reading adults. Within a monotonous sequence of 50-millisecond 1000 Hz binaural tones, tones of 920 and 1080 Hz occurred with 7% probability each. Magnetic mismatch fields, elicited by the stimulus deviances, were diminished in the left hemisphere of the dyslexic subjects. The results indicate deficient change detection in the left auditory cortex of right-handed dyslexic adults. PMID- 12730989 TI - Randomized trial of pallidotomy versus medical therapy for Parkinson's disease. AB - Thirty-six patients with Parkinson's disease (PD) were randomized to either medical therapy (N = 18) or unilateral GPi pallidotomy (N = 18). The primary outcome variable was the change in total Unified Parkinson's Disease Rating Scale (UPDRS) score at 6 months. Secondary outcome variables included subscores and individual parkinsonian symptoms as determined from the UPDRS. At the six month follow-up, patients receiving pallidotomy had a statistically significant reduction (32% decrease) in the total UPDRS score compared to those randomized to medical therapy (5% increase). Following surgery, patients' showed improvement in all the cardinal motor signs of PD including tremor, rigidity, bradykinesia, gait and balance. Drug-induced dyskinesias were also markedly improved. Although the greatest improvement occurred on the side contralateral to the lesion, significant ipsilateral improvement was also observed for bradykinesia, rigidity and drug-induced dyskinesias. A total of twenty patients have been followed for 2 years to assess the effect of time on clinical outcome. These patients have shown sustained improvement in the total UPDRS (p < 0.0001), "off" motor (p < 0.0001) and complications of therapy subscores (p < 0.0001). Sustained improvement was also seen for tremor, rigidity, bradykinesia, percent on time and drug-induced dyskinesias. PMID- 12730990 TI - Anti-GM1 IgG antibodies induce leukocyte effector functions via Fcgamma receptors. AB - Guillain-Barre syndrome (GBS) is an immune-mediated neuropathy, in which leukocytes and humoral components of the immune system proposedly initiate localized inflammation. An important pathogenic role for anti-GM1 ganglioside antibodies has been suggested. Therefore, we evaluated anti-GM1 IgG antibody induced leukocyte effector functions such as degranulation and phagocytosis using serum of 24 GBS patients. Serum without anti-GM1 antibodies of 9 GBS patients as well as pooled serum from healthy individuals served as controls. Ten out of 15 (67%) of anti-GM1 IgG positive sera were capable of inducing leukocyte degranulation, and 8 out of 15 (53%) of anti-GM1 IgG positive sera were capable of inducing phagocytosis of GM1-coated beads. In all of these sera anti-GM1 antibody titers were >or=1:800. No leukocyte degranulation or phagocytosis was observed in control sera. Leukocyte activation was completely abrogated in the presence of IgG receptor (FcgammaR) blocking antibodies, suggesting a crucial role for leukocyte FcgammaR in GBS pathogenesis. No correlation of antibody titers with the extent of leukocyte activation, or severity of disease was observed. These data document the capacity of anti-GM1 IgG antibodies to activate leukocyte inflammatory functions, and suggest an important role for anti ganglioside IgG antibodies in the pathogenesis of GBS. PMID- 12730991 TI - Anti-neuronal nuclear autoantibody type 2: paraneoplastic accompaniments. AB - We identified the IgG autoantibody ANNA-2 ("anti-Ri") in 34 patients in a 12-year period by immunofluorescence screening of sera from approximately 75000 patients with subacute neurological disorders that were suspected to be paraneoplastic. Detailed clinical information was available for 28 patients (10 men, 18 women). Cancer was diagnosed in 24 patients (86%); 21 had histologically proven carcinoma (10 lung, 9 breast, 1 cervical, 1 bladder), and 3 had an intrathoracic imaging abnormality. Cancer anteceded neurological symptoms in 4 of 28 patients. Cancer detection frequency increased with continued surveillance. Neurological disorders, in decreasing frequency, were brainstem syndrome (including opsoclonus, myoclonus, or both), cerebellar syndrome, myelopathy, peripheral neuropathy, cranial neuropathy, movement disorder, encephalopathy, Lambert-Eaton syndrome, and seizures. Four patients had laryngospasm and four had jaw opening dystonia (two with neck dystonia). Nine (32%) were wheelchair-bound 1 month after neurological symptom onset. Most improved neurologically after immunomodulatory or tumor-directed therapy. Accompanying autoantibodies, found in 73% of sera, included ANNA-1, ANNA-3, CRMP-5-IgG, P/Q-type and N-type Ca(2+) channel antibodies, and muscle-type acetylcholine receptor antibody. Some neurological accompaniments of ANNA-2 may reflect potentially pathogenic humoral or cell mediated responses to coimmunogenic tumor antigens, for example, Lambert-Eaton syndrome (P/Q-type Ca(2+) channel antibody) and peripheral neuropathy (ANNA-1 effector T cells). PMID- 12730992 TI - Interleukin-1beta promotes oligodendrocyte death through glutamate excitotoxicity. AB - Glutamate excitotoxicity is implicated in the progressive loss of oligodendrocytes in multiple sclerosis, but how glutamate metabolism is dysregulated in the disease remains unclear. Because there is microglia activation in all stages of multiple sclerosis, we determined whether a microglia product, interleukin-1beta, could provide the mechanism for glutamate excitotoxicity. We found that whereas interleukin-1beta did not kill oligodendrocytes in pure culture, it produced apoptosis of oligodendrocytes in coculture with astrocytes and microglia. This requirement for a mixed glia environment suggests that interleukin-1beta impairs the well-described glutamate buffering capacity of astrocytes. In support, antagonists at AMPA/kainate glutamate receptors, NBQX and CNQX, blocked the interleukin-1beta toxicity to oligodendrocytes. Another microglia/macrophage cytokine, tumor necrosis factor alpha, also evoked apoptosis of oligodendrocytes in a mixed glia environment in an NBQX-blockable manner. These results provide a mechanistic link between the persistent and insidious microglia activation that is evident in all stages of multiple sclerosis, with the recent appreciation that glutamate excitotoxicity leads to the destruction of oligodendrocytes in the disease. PMID- 12730993 TI - Bilateral frontoparietal polymicrogyria: clinical and radiological features in 10 families with linkage to chromosome 16. AB - Polymicrogyria is a common malformation of cortical development characterized by an excessive number of small gyri and abnormal cortical lamination. Multiple syndromes of region-specific bilateral symmetric polymicrogyria have been reported. We previously have described two families with bilateral frontoparietal polymicrogyria (BFPP), an autosomal recessive syndrome that we mapped to a locus on chromosome 16q12-21. Here, we extend our observations to include 19 patients from 10 kindreds, all linked to the chromosome 16q locus, allowing us to define the clinical and radiological features of BFPP in detail. The syndrome is characterized by global developmental delay of at least moderate severity, seizures, dysconjugate gaze, and bilateral pyramidal and cerebellar signs. Magnetic resonance imaging demonstrated symmetric polymicrogyria affecting the frontoparietal regions most severely, as well as ventriculomegaly, bilateral white matter signal changes, and small brainstem and cerebellar structures. We have refined our genetic mapping and describe two apparent founder haplotypes, one of which is present in two families with BFPP and associated microcephaly. Because 11 of our patients initially were classified as having other malformations, the syndrome of BFPP appears to be more common than previously recognized and may be frequently misdiagnosed. PMID- 12730994 TI - Endogenous estradiol and risk of dementia in women and men: the Rotterdam Study. AB - We determined whether higher endogenous estradiol levels were associated with lower risk of dementia in older men and women not using hormonal replacement therapy, using a case-cohort design within the Rotterdam Study, a population based follow-up study on chronic diseases, including dementia, in 7983 subjects aged 55 years or older, and ongoing since 1990. The analyses were based on a random subcohort of 508 women and 438 men, and on 76 women and 53 men with incident dementia. Cox proportional hazards models with robustly estimated standard errors showed that in women higher levels of total estradiol were associated with higher risk of dementia (age-adjusted hazard ratio per standard deviation increase 1.38; 95% CI 1.04-1.84). Age-adjusted HR's of Alzheimer's disease and vascular dementia associated with higher levels of total estradiol (per SD increase) were 1.24 (95% CI 0.87-1.76) and 2.19 (95% CI 1.22-3.92), respectively. Similar results were observed for bioavailable estradiol. Additional adjustments for potential confounders did not change the results substantially. In men, no clear association was observed between estradiol levels and risk of dementia or its subtypes. The findings do not support the hypothesis that higher levels of endogenous estradiol reduce risk of dementia, neither in women nor in men. PMID- 12730995 TI - Tremor and ataxia in fragile X premutation carriers: blinded videotape study. AB - Fragile X premutation carriers do not have typical fragile X syndrome (FXS) although late-onset progressive action tremor and gait disorder with CNS atrophy was recently reported in male carriers. We compared tremor, gait disorder and parkinsonian signs in FXS premutation subjects (age 50 or more) and a similar control population, using a standardized videotaping protocol. Videotapes were rated using standard scales for tremor (CRST), ataxia (ICARS), and parkinsonian signs (UPDRS) by an investigator blinded to premutation status. Compared to all other groups pooled (n = 30), the male premutation carrier group (n = 7) had significantly higher scores on the CRST (p = 0.0008), ICARS (p = 0.001), and UPDRS (p = 0.0094). On the CRST, rest, postural and kinetic tremor scores were all higher in the male carriers. The elevated total UPDRS and ICARS scores mainly resulted from markedly higher scores for tremor and limb ataxia, respectively. The female carrier (n = 14) and control groups (n = 8) did not differ on any measure. The FMR1 premutation is associated with increased levels of CGG repeat containing FMR1 mRNA, which may predispose to these symptoms by interfering with nuclear mechanisms. Given the relatively high population frequency of the FMR1 premutation, this mutation may be a significant cause of late-onset "idiopathic" progressive tremor. PMID- 12730997 TI - Temporal processing of visuotactile and tactile stimuli in writer's cramp. AB - Neurophysiological studies in animals show that basal ganglia are involved not only in motor and nonmotor timing functions but also in integrating tactile and visual signals delivered in the peripersonal space. We tested temporal discrimination of cross-modal and unimodal stimuli in 13 controls and 14 patients with writer's cramp, a disorder supposedly linked to dysfunction of basal ganglia. Subjects were asked to discriminate whether pairs of visual, tactile, or visuotactile stimuli were simultaneous or sequential (temporal discrimination threshold) and which stimulus preceded the other (temporal order judgment). Patients were impaired in temporal processing of tactile and cross-modal stimuli. A significant positive correlation between temporal deficits and the severity of disability was detected for both affected and unaffected sides. Findings suggest that multimodal and not only modality-specific temporal processing is defective in focal hand dystonia. Deficits of temporal processing of stimuli delivered to the unaffected side may represent a behavioral index of the susceptibility to develop dystonia and thus have remarkable practical and theoretical implications. PMID- 12730996 TI - Parkin mutations and susceptibility alleles in late-onset Parkinson's disease. AB - Parkin, an E2-dependent ubiquitin protein ligase, carries pathogenic mutations in patients with autosomal recessive juvenile parkinsonism, but its role in the late onset form of Parkinson's disease (PD) is not firmly established. Previously, we detected linkage of idiopathic PD to the region on chromosome 6 containing the Parkin gene (D6S305, logarithm of odds score, 5.47) in families with at least one subject with age at onset (AAO) younger than 40 years. Mutation analysis of the Parkin gene in the 174 multiplex families from the genomic screen and 133 additional PD families identified mutations in 18% of early-onset and 2% of late onset families (5% of total families screened). The AAO of patients with Parkin mutations ranged from 12 to 71 years. Excluding exon 7 mutations, the mean AAO of patients with Parkin mutations was 31.5 years. However, mutations in exon 7, the first RING finger (Cys253Trp, Arg256Cys, Arg275Trp, and Asp280Asn) were observed primarily in heterozygous PD patients with a much later AAO (mean AAO, 49.2 years) but were not found in controls in this study or several previous reports (920 chromosomes). These findings suggest that mutations in Parkin contribute to the common form of PD and that heterozygous mutations, especially those lying in exon 7, act as susceptibility alleles for late-onset form of Parkinson disease. PMID- 12730998 TI - Lineage pathway of human brain progenitor cells identified by JC virus susceptibility. AB - Multipotential human central nervous system progenitor cells, isolated from human fetal brain tissue by selective growth conditions, were cultured as undifferentiated, attached cell layers. Selective differentiation yielded highly purified populations of neurons or astrocytes. This report describes the novel use of this cell culture model to study cell type-specific recognition of a human neurotropic virus, JC virus. Infection by either JC virions or a plasmid encoding the JC genome demonstrated susceptibility in astrocytes and, to a lesser degree, progenitor cells, whereas neurons remained nonpermissive. JC virus susceptibility correlated with significantly higher expression of the NFI-X transcription factor in astrocytes than in neurons. Furthermore, transfection of an NFI-X expression vector into progenitor-derived neuronal cells before infection resulted in viral protein production. These results indicate that susceptibility to JC virus infection occurs at the molecular level and also suggest that differential recognition of the viral promoter sequences can predict lineage pathways of multipotential progenitor cells in the human central nervous system. PMID- 12730999 TI - Endogenous dopamine release after pharmacological challenges in Parkinson's disease. AB - Using (11)C-raclopride positron emission tomography after methamphetamine challenge, we have evaluated regional brain changes in synaptic dopamine (DA) levels in six volunteers and six advanced Parkinson's disease (PD) patients. The pharmacological challenge induced significant release of endogenous DA in putamen not only in the normal subjects, as reflected by a 25.2% reduction in (11)C raclopride binding potential as compared with placebo, but also in the PD patients (6.8%). In individual PD patients, we found a correlation between putamen DA release and DA storage, as measured by (18)F-dopa uptake. Localization of significant changes in (11)C-raclopride binding after methamphetamine at a voxel level with statistical parametric mapping identified striatal and prefrontal DA release in both cohorts. Statistical comparisons between normal subjects and PD confirmed significantly reduced DA release in striatal areas in PD, but normal levels of prefrontal DA release. In conclusion, significant endogenous DA release can still be induced by pharmacological challenges in the putamen of advanced PD patients, and this release correlates with residual DA storage capacity. Our data also show that the capacity to release normal DA levels in prefrontal areas after a pharmacological challenge is preserved in severe stages of the disease. PMID- 12731000 TI - Decoupling of autonomic and cognitive emotional reactions after cerebellar stroke. AB - Emotional blunting can be found after cerebellar lesions. However, the mechanism of such a modification is not clear. We present a patient with emotional flattening and increased risk taking after left cerebellar infarct who had an impaired autonomic reactivity to negative as compared with positive reinforcement. This impairment was demonstrated by the patient's undifferentiated skin conductance responses to negative and positive reinforcement, whereas controls produced larger skin conductance responses after negative feedback. The cooccurrence of emotional flattening and undifferentiated autonomic reactions to positively and negatively valenced stimuli strengthens the role of the cerebellum in the modulation of the autonomic responses. PMID- 12731001 TI - Polymicrogyria and absence of pineal gland due to PAX6 mutation. AB - Identification of genes involved in human cerebral development is important for our understanding of disorders with potential neurodevelopmental causes such as epilepsy and learning disability. Murine models suggest that PAX6 plays a key role in human brain development. With magnetic resonance imaging in 24 humans heterozygous for defined PAX6 mutations, we demonstrated widespread structural abnormalities including absence of the pineal gland and unilateral polymicrogyria. PMID- 12731002 TI - Functional magnetic resonance imaging of human absence seizures. AB - We studied a patient with idiopathic generalized epilepsy and frequent absences, using electroencephalogram-correlated functional magnetic resonance imaging. Four prolonged runs of generalized spike-wave discharge occurred during a 35-minute experiment. Time-locked activation was observed bilaterally within the thalami in conjunction with widespread but symmetrical cortical deactivation with a frontal maximum. We demonstrate the reciprocal participation of focal thalamic and widespread cortical networks during human absence seizures and suggest reductions in cortical blood flow, in response to synchronized electroencephalogram activity. PMID- 12731003 TI - Markers of cellular proliferation are expressed in cortical tubers. AB - p34cdc2, collapsin response mediator protein 4 (CRMP4), doublecortin (DCX), HuD, and NeuN expression was assessed in tuber (n = 16) and subependymal giant cell astrocytoma (SEGA; n = 6) specimens in tuberous sclerosis complex to define the developmental phenotype and lineage of giant cells (CGs) in these lesions. Many GCs exhibited HuD and NeuN immunolabeling suggesting a differentiated neural phenotype. Giant cells in tubers, SEGAs and subependymal nodules in the Eker rat model of TSC expressed CRMP4 and DCX. Tubers and SEGAs exhibit a heterogeneous profile of differentiation and may share a common cellular lineage. Tubers may contain a subpopulation of newly generated cells. PMID- 12731005 TI - Weight loss in Parkinson's disease. AB - We prospectively examined the changes of body weight among 468 individuals with Parkinson's disease (PD) before and after the diagnosis. The average weight of PD patients was stable until shortly before the diagnosis (p for trend = 0.2) and then declined (p for trend < 0.0001). Interestingly, PD patients tended to increase their energy intakes as their weights decreased (p for trend < 0.002). In conclusion, weight loss in PD patients appears to be a continuous process that starts several years before the diagnosis and is not caused by reduced energy intake. PMID- 12731004 TI - New T2 lesions enable an earlier diagnosis of multiple sclerosis in clinically isolated syndromes. AB - In clinically isolated syndromes, the new McDonald criteria for multiple sclerosis diagnosis require new gadolinium-enhancing lesions for dissemination in time at a 3-month follow-up magnetic resonance imaging scan. In a cohort of 56 patients, these criteria were specific (95%) but less sensitive (58%) for clinically definite multiple sclerosis at 3 years. If new T2 lesions were allowed as an alternative for dissemination in time, sensitivity increased (74%) with maintained specificity (92%), enabling an accurate diagnosis of multiple sclerosis in more patients. PMID- 12731006 TI - Efficient central nervous system remyelination requires T cells. AB - We demonstrate a role for immune functions in the spontaneous remyelination of central nervous system (CNS) axons after lysolecithin-induced demyelination in the spinal cord. Rag-1-deficient mice lack both B cells and T cells and show significantly reduced spontaneous remyelination compared with control mice of matching genetic background. Mice lacking or depleted of either CD4(+) T cells or CD8(+) T cells also exhibit reduced remyelination. These data indicate that T cells are necessary for efficient CNS remyelination. Thus, general nonspecific immunosuppression as a therapeutic approach for the treatment of CNS injury and demyelinating disease may have undesirable effects on subsequent tissue repair. PMID- 12731008 TI - Concerns regarding transience and heterozygosity in neonatal hyperglycenemia. PMID- 12731009 TI - Is there mitochondrial dysfunction in amyotrophic lateral sclerosis skeletal muscle? PMID- 12731011 TI - Valporic acid and prion proteins. PMID- 12731012 TI - Clinical significance of positionally induced downbeat nystagmus. PMID- 12731017 TI - Delivery of bioactive, gel-isolated proteins into live cells. AB - The delivery of proteins into live cells is a promising strategy for the targeted modulation of protein-protein interactions and the manipulation of specific cellular functions. Cellular delivery can be facilitated by complexing the protein of interest with carrier molecules. Recently, an amphipatic peptide was identified, Pep-1 (KETWWETWWTE WSQPKKKRKV), which crosses the plasma membrane of many cell types to carry and deliver proteins as large as antibodies. Pep-1 effectively delivers proteins in solution; but Pep-1 is not suitable for delivering sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) isolated proteins because Pep-1 complexes with cargo proteins are destroyed by SDS. Here, we report cellular delivery of SDS-PAGE-isolated proteins, without causing cellular damage, by using a nonionic detergent, Triton X-100, as carrier. To determine the specificity of our method, we separated antibodies against different intracellular targets by nonreducing SDS-PAGE. Following electrophoresis, the antibody bands were detected by zinc-imidazole reverse staining, excised, in-gel refolded with Triton X-100, and eluted in detergent free phosphate-buffered saline. When overlaid on cultured NIH 3T3 cells, the antibodies penetrated the cells localizing to their corresponding intracellular targets. These results are proof-of-principle for the delivery of gel-isolated bioactive proteins into cultured cells and suggest new ways for experimental protein therapy and for studying protein-protein interactions using gel-isolated protein. PMID- 12731018 TI - Electrophoresis of biological cells: charge-regulation and multivalent counterions association model. AB - The electrophoresis of a biological cell is analyzed theoretically. An entity, which is of amphoteric nature, is used to simulate its electrophoretic behavior. To reflect conditions of practical interest, we assume that the liquid phase contains mixed (a:b)+(c:b) electrolytes, where a and c are the valences of cations, and b is the valence of anions. We consider the case where the surface of a cell contains both bivalent acidic and monovalent basic functional groups, the dissociation/association of them yields fixed surface charge, and the multivalent cations in the liquid phase are allowed to combine with dissociated acidic functional groups, which has the effect of lowering the charge density on cell surface. The electrophoretic behaviors of a cell under various conditions are illustrated. The results obtained can be used to identify the types of functional groups that may be present on cell surface. On the other hand, if the surface functional groups involved in cell electrophoresis are known, then their density and the associated dissociation/association constants can be estimated from experimental data. PMID- 12731019 TI - Direct and specific recognition of glycosaminoglycans by antibodies after their separation by agarose gel electrophoresis and blotting on cetylpyridinium chloride-treated nitrocellulose membranes. AB - A method for the immunodetection of several natural complex polysaccharides (glycosaminoglycans) after their separation by conventional agarose gel electrophoresis, blotting and immobilizing on nitrocellulose membranes derivatized with the cationic detergent cetylpyridinium chloride (CPC), and direct and specific immunodetection by antibodies is described. This new approach is based on the principles that were used to develop the Western blot, and is applied to the separation of the glycosaminoglycans purified from normal human urine. After migration in agarose gel electrophoresis, chondroitin sulfate samples of different origin were blotted and transferred onto nitrocellulose membranes treated with CPC. Immunodetection was performed using the anti chondroitin-6-sulfate antibody that specifically recognizes intact chondroitin-6 sulfate. By calculating the ratio between the antibody staining (epitope) and alcian blue staining (mass), the epitope density expressed as a percentage, i.e., the number of repetitive epitopes per mass, was obtained. These values were in agreement with the quantitation of 6-sulfated groups of chondroitin sulfate performed by the evaluation of unsatured disaccharide-6-sulfate (DeltaDi6S) produced after treatment with chondroitinase ABC and separated by high performance liquid chromatography (HPLC). Furthermore, immunodetection of heparan sulfate was performed using the anti-heparan sulfate antibody. PMID- 12731020 TI - Genetic differentiation and subspecies development of the giant panda as revealed by DNA fingerprinting. AB - Over the last 100 years giant pandas (Ailuropoda melanoleuca) have been separated into six completely isolated mountain ranges. DNA fingerprinting revealed different differentiation patterns in giant pandas, including early-stage, late stage, recent divergence and recent genetic depression. A separation around 10 000 years ago resulted in highly significant differences in DNA fingerprints and morphological characteristics between Qinling and Sichuan populations. Supported by morphological differences, the genetic data were used to classify the Qinling population as a new subspecies, A. m. qinlingensis, while the Sichuan populations were classified into the original subspecies, A. m. melanoleuca. Thus, the Qinling population deserves management as a separate unit. In the Sichuan populations, two management units were defined, including Qionglai-Minshan and Daxiangling-Xiaoxiangling-Liangshan. Our data suggest urgent measures are needed to establish green corridors between subpopulations in each mountain range to increase gene flow and genetic variation to ensure long-term survival. PMID- 12731021 TI - Chiral resolution of some environmental pollutants by capillary electrophoresis. AB - The chiral resolution of the environmental pollutants by capillary electrophoresis is reviewed. Various aspects of the chiral resolution such as chiral selectors, optimization of capillary electrophoresis conditions including composition of the background electrolyte (BGE), pH of the BGE, ionic strength of the BGE, structures and types of the chiral selectors, applied voltage, temperature, structures of the chiral pollutants, use of organic modifiers and other parameters are presented. Furthermore, detection, sample treatment, validation of the methods, and the chiral recognition mechanisms, have been discussed. PMID- 12731022 TI - Effect of buffer, electric field, and separation time on detection of aptamer ligand complexes for affinity probe capillary electrophoresis. AB - The separation and detection of complexes of aptamers and protein targets by capillary electrophoresis (CE) with laser-induced fluorescence was examined. Aptamer-thrombin and aptamer-immunoglobulin E (IgE) were used as model systems. Phosphate, 3-(N-morpholino)propanesulfonic acid with phosphate, and tris(hydroxyamino)methane-glycine-potassium (TGK) buffer at pH 8.4 were tested as electrophoresis media. Buffer had a large effect with TGK providing the most stable complexes for both protein-aptamer complexes. Conditions that suppressed electroosmotic flow, such as addition of hydroxypropylmethylcellulose to the media or modification of the capillary inner wall with polyacrylamide, were found to prevent detection of complexes. The effect of separation time and electric field were evaluated by monitoring complexes with electric field varied from 150 2850 V/cm and effective column lengths of 3.5 and 7.0 cm. As expected, shorter times on the column greatly increased peak heights for the complexes due to a combination of less dilution by diffusion and less dissociation on the column. High fields were found to have a detrimental effect on detection of complexes. It is concluded that the best conditions for detection of noncovalent complexes involve use of the minimal column length and electric field necessary to achieve separation. The results will be of interest in developing affinity probe CE assays wherein aptamers are used as affinity ligands. PMID- 12731023 TI - Sol-gel column technology for capillary isoelectric focusing of microorganisms and biopolymers with UV or fluorometric detection. AB - The sol-gel surface modification is used for capillary isoelectric focusing of microorganisms and biopolymers. The coating strongly decreases the electroosmotic flow so that it enables the use of the short capillaries down to 100 mm in the separation length. The examples of capillary isoelectric focusing of the low molecular-mass pI markers and mixed cultures of microbial populations of Escherichia coli, Candida albicans, Staphylococcus epidermidis, and Enteroccocus faecalis with UV detection are shown. It is possible to quantify bacterial cells according to their peak areas; the minimum detectable number of microbial cells was 5 x 10(2)-1 x 10(3). The compatibility of sol-gel capillaries with sensitive fluorometric detection of fluorescent pI markers together with fluorescein labeled proteins is demonstrated. PMID- 12731024 TI - Size-based separations of proteins by capillary electrophoresis using linear polyacrylamide as a sieving medium: model studies and analysis of cider proteins. AB - Electrophoretic conditions to separate sodium dodecyl sulfate (SDS)-protein complexes according to their relative molecular mass by capillary electrophoresis (CE) using linear polyacrylamide as a sieving matrix were examined. Five purified proteins with relative molecular masses between 14 400 and 66 200 Da were separated on a coated fused-silica capillary with an internal diameter of 100 microm and an effective length of 24 cm (total length, 32.5 cm). Benzoic acid was added to the solution of purified proteins as internal standard; beta mercaptoethanol was also added as reducing agent. The running buffer composition was 0.05 M tris(hydroxymethyl)aminomethane (Tris), 0.035 M aspartic acid, 0.1% m/v SDS, 4% m/v acrylamide, the resulting pH being 8.0. The applied voltage was 7 kV (reversed voltage polarity) in order to avoid high current intensities. Under optimized conditions, the five proteins were separated in less than 15 min, with a % relative standard deviation (RSD) between 0.2 and 0.4 for migration times in the same day. Good efficiency (values between 150 000 and 40 000 N/m) and resolution (values between 2 and 2.8) were obtained. The inverse of relative migration times was found to correlate with the logarithm of their relative molecular mass. Finally, cider proteins were analyzed and their relative molecular masses were determined. These results were compared with those obtained by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). PMID- 12731025 TI - Speciation of chromium (III) and chromium (VI) by capillary electrophoresis with contactless conductometric detection and dual opposite end injection. AB - A sensitive, rapid and inexpensive capillary electrophoretic method for the determination of Cr(III) and Cr(VI) species is presented. The method is based on the dual opposite end injection principle and contactless conductometric detection. The sample containing cationic and anionic species is injected into the opposite ends of the separation capillary and after the high voltage is applied, the analytes migrate towards the capillary center, where the cell of a contactless conductivity detector is placed. The method does not require any sample pretreatment, except dilution with deionized water. The separation of Cr(III), Cr(VI) and other common inorganic anions and cations is achieved in less than 4 min. The parameters of the separation electrolyte solution, such as pH and concentration of L-histidine, were optimized. Best results were achieved with electrolyte solution consisting of 4.5 mM L-histidine, adjusted to pH 3.40 with acetic acid. The detection limits achieved for Cr(III) and Cr(VI) were 10 and 39 microg.L(-1), respectively. The repeatability of migration times and peak areas was better than 0.3% and 2.8%, respectively. The developed method was applied to the analyses of rinse water samples from the galvanic industry. The results for the determination of Cr(III) and Cr(VI) were in good agreement with the results obtained by certified differential spectrophotometric method using diphenylcarbazide (CN 83 0520-40) and with the results for the total chromium concentrations determined by electrothermal atomic absorbance spectrometry (ET AAS) and inductively coupled plasma-mass spectrometry (ICP-MS). PMID- 12731026 TI - Micellar electrokinetic chromatography estimation of size and composition of procyanidins after thiolysis with cysteine. AB - This paper describes the characterization of procyanidin mixtures by acid depolymerization in the presence of cysteine (thiolysis with cysteine) and micellar electrokinetic chromatography (MEKC). Reversed-phase liquid chromatography (RP-HPLC) and MEKC were investigated for the separation of the major components of the depolymerized mixtures (catechins and their cysteinyl derivatives). The solutes could only be effectively separated using MEKC. Two background electrolytes (BGEs) are recommended: (i) 50 mM phosphate at pH 7, containing 40 mM sodium cholate (SC) and 10 mM sodium dodecyl sulfate (SDS); (ii) a BGE with the same composition but containing only 50 mM SDS. The MEKC procedures here reported, are cheap, reliable and fast, and their potential in the determination of the size and composition in procyanidin mixtures has been shown. The proposed MEKC methods were validated by comparison with our intralaboratory reference RP-HPLC method using cysteamine as thiol donor. PMID- 12731027 TI - Capillary electrophoresis with end-column amperometric detection of urinary 8 hydroxy-2'-deoxyguanosine. AB - Urinary 8-hydroxy-2'-deoxyguanosine (8OHdG) is an excellent marker of oxidative DNA damage. Until now, urinary 8OHdG has been measured by high-performance liquid chromatography with electrochemical detection. A simple and sensitive method for the analysis of urinary 8OHdG by capillary electrophoresis with end-column amperometric detection has been developed in our laboratory. A single-step solid phase extraction procedure was optimized and used for extracting 8OHdG from human urine. To improve the sensitivity of this method, a new focusing technique based on a dynamic pH junction was used. The limit of detection was 20 nM (signal-to noise ratio S/N = 3), the linear range was 50 nM-10 microM, and the correlation coefficient was better than 0.999. The relative standard deviation (RSD) was found to be 0.57% for migration time, and 4.79% for peak current. To show the usefulness of the method, the urinary concentration of 8OHdG in nine healthy persons and ten cancer patients was determined. The urinary concentration of 8OHdG in cancer patients was significantly higher than that in healthy persons. PMID- 12731028 TI - Determination of enantiomeric purity of a novel COX-2 anti-inflammatory drug by capillary electrophoresis using single and dual cyclodextrin systems. AB - E-6087 is the most advanced compound among the cyclooxygenase-2 (COX-2) inhibitor drugs developed in our company. Its activity is mainly associated with the S(-) enantiomer (E-6232), whereas the R(-)-enantiomer (E-6231) becomes an impurity whose content should be determined. Five main impurities and degradation products of E-6232 have been found (E-6144, E-6024, E-6072, E-6397 and E-6132), and some of them co-elute with the distomer when using a chiral high-performance liquid chromatography (HPLC) method. Consequently, we have optimized the separation of all the impurities from the two enantiomers of E-6087 by capillary electrophoresis (CE), in order to use the method for the enantiomeric purity determination of E-6232. The effect of the methanol (MeOH) content in the background electrolyte (BGE), the sulfobutyl ether-beta-cyclodextrin (SBE-beta CD) and heptakis-(2,6-di-O-methyl)-beta-cyclodextrin (DM-beta-CD) concentration, and the capillary temperature have been studied. Separation of all compounds could be achieved in different systems, either in a single CD-system (with SBE beta-CD) or in a dual CD-system (with DM-beta-CD as a neutral CD). By using the dual CD system a limit of detection (LOD) and a limit of quantitation (LOQ) of 0.03% and 0.1% of distomer, respectively, were achieved*. PMID- 12731029 TI - Investigation of a capillary electrophoretic approach for direct quantification of apolipoprotein A-I in serum. AB - In the present study a rapid, reproducible and robust capillary electrophoresis (CE) procedure for the quantification of apolipoprotein A-I (Apo A-I) in serum without pretreatment has been developed (total run time, 11 min). The coefficients of variation (CV; n = 10) for the relative peak area are 1.8% at a concentration of 145 mg/dL and 1.6% at 196 mg/dL; and for the inter-assay 8.9% at 161 mg/dL (10 consecutive days), i.e., similar to the CVs of a high-throughput immunonephelometric routine assay. The CV for the migration time is 0.4% (n = 20). The robustness of the CE approach was tested in patient samples with hemolysis, hyperbilirubinemia and hyperlipidemia. A comparison of 99 Apo A-I serum values with results of a fixed-time immunonephelometric routine assay showed a positive constant bias of 60% (mean) for the immunonephelometric values, no deviation from linearity, but significant deviations in several samples. Investigations on interferences in the CE analyses gave no evidence that CE failed. Our study shows that CE is amenable to a fast analysis and a reproducible and reliable quantification of Apo A-I level in sera of various clinical samples. PMID- 12731030 TI - Capillary electrophoresis separation of high molecular weight glutenin subunits in bread wheat (Triticum aestivum L.) and related species with phosphate-based buffers. AB - This study focused on optimizing phosphate-based buffers and other capillary electrophoresis (CE) parameters for separating and characterizing high molecular weight glutenin subunits (HMW-GS) in bread wheat (Triticum aestivum L., AABBDD, 2n = 6x = 42), emmer (Triticum dicoccum, AABB, 2n = 4x = 28) and Aegilops tauschii (DD, 2n = 2x = 14). The fast and high-resolution separation of HMW-GS was achieved using 0.1 M phosphate-glycine buffer (pH 2.5, containing 20% acetonitrile and 0.05% hydroxypropylmethylcellulose) at 12.5 kV and 40 degrees C with 25 microm inside diameter (ID)x27 cm uncoated fused-silica capillary. In general, one sample separation can be analyzed in 15 min. The good run-to-run repeatable separation of HMW-GS could be obtained with a relative standard deviation of less than 1% when capillaries were rinsed with 1 M phosphoric acid for 2 min, followed by separation buffer for 2 min after each separation. The HMW GS from some bread wheat cultivars as well as tetraploid and diploid accessions was separated by the CE method described above, and all subunits detected were well characterized and readily identified. Some HMW-GS showed reversed mobilities and elution order compared to the methods of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and SDS-CE. Particularly, most of the HMW-GS analyzed with the CE buffer used were separated into multiple peaks, generally a high peak plus a minor peak. CE appears to be capable of separating and characterizing HMW-GS with fast and high-resolution features, therefore it is expected to be useful for specific germplasm screening and desirable HMW-GS identification in wheat quality improvement. PMID- 12731032 TI - Surface characterization using chemical force microscopy and the flow performance of modified polydimethylsiloxane for microfluidic device applications. AB - The widespread interest in micro total analysis systems has resulted in efforts to develop devices in cheaper polymer materials such as polydimethylsiloxane (PDMS) as an alternative to expensive glass and silicon devices. We describe the oxidation of the PDMS surface to form ionizable groups using a discharge from a Tesla coil and subsequent chemical modification to augment electroosmotic flow (EOF) within the microfluidic devices. The flow performance of oxidized, amine modified and unmodified PDMS materials has been determined and directly compared to conventional glass devices. Exact PDMS replicas of glass substrates were prepared using a novel two step micromolding protocol. Chemical force microscopy has been utilized to monitor and measure the efficacy of surface modification yielding information about the acid/base properties of the modified and unmodified surfaces. Results with different substrate materials correlates well with expected flow modifications as a result of surface modification. Oxidized PDMS devices were found to support faster EOF (twice that of native PDMS) similar to glass while those derivatized with 3-aminopropyl triethoxysilane (APTES) showed slower flow rates compared to native PDMS substrates as a result of masking surface charge. Results demonstrate that the surface of PDMS microdevices can be manipulated to control EOF characteristics using a facile surface derivatization methodology allowing surfaces to be tailored for specific microfluidic applications and characterized with chemical force microscopy. PMID- 12731031 TI - Application of capillary zone electrophoresis in the separation and determination of the principal gum opium alkaloids. AB - We describe the use of capillary zone electrophoresis (CZE) for the qualitative and quantitative determination of major alkaloids (i.e., thebaine, codeine, morphine, papavarine and narcotine) in gum opium involving the analysis of alkaloids without derivatization or purification. Three extractions with 2.5% w/v aqueous acetic acid quantitatively extracted major alkaloids. The separation was carried out by CZE using a 7:3 mixture of methanol and sodium acetate (100 mM, pH 3.1) at a potential of 15 kV, with UV detection at 224 nm. Spiking of pure reference alkaloid standards in the opium extract was used for peak identification. The influences of buffer composition, pH and voltage on the separation of alkaloids were studied. The detection limit of each alkaloid dissolved in methanol was found to be 850 ng/mL (morphine), 450 ng/mL (thebaine), 500 ng/mL (codeine), 550 ng/mL (papaverine), and 500 ng/mL (narcotine) at an injection pressure of 300 mbar (injection volume, 4 nL) with a signal-to-noise ratio of 3:1. The external standard method was used for the quantification of alkaloids. The calibration plot was based on linear regression analysis. The relative standard deviation (RSD) for peak area and migration time was in the range of 1.03-3.56% and 0.34-0.69%, respectively. Percentage compositions (g%) of opium alkaloids in five gum opium samples were found to be in the range of 14.45 15.95 (morphine), 2.0-3.45 (codeine), 1.32-2.73 (thebaine), 0.92-2.37 (papavarine), and 3.85-5.77 (narcotine). The method developed is suitable for the routine analysis of major gum opium alkaloids in samples of forensic importance. PMID- 12731033 TI - Comprehensive two-dimensional separations based on capillary high-performance liquid chromatography and microchip electrophoresis. AB - A novel comprehensive two-dimensional (2-D) separation system coupling capillary high-performance liquid chromatography (cHPLC) with microchip electrophoresis (chip CE) is demonstrated. Reversed-phase cHPLC was used as the first dimension, and chip CE acted as the second dimension to perform fast sample transfers and separations. A valve-free gating interface was devised simply by inserting the outlet-end of LC column into the cross-channel on a specially designed chip. A home-made confocal laser-induced fluorescence detector was used to perform on chip high-sensitive detection. The cHPLC effluents were continuously delivered to the chip and pinched injections of the effluents every 20 seconds were employed for chip CE separation. Gradient elution of cHPLC was carried out to obtain the high-efficiency separation. Free-zone electrophoresis was performed with triethylamine buffer to achieve high-speed separation and prevent sample adsorption. Such a simple-made comprehensive system was proved to be effective. The relative standard deviations for migration time and peak height of rhodamine B in 150 sample transfers were 3.2% and 9.8%, respectively. Peptides of the fluorescein isothiocyanate (FITC)-labeled tryptic digests of bovine serum albumin were fairly resolved and detected with this comprehensive 2-D system. PMID- 12731034 TI - Proteome analysis of Saccharomyces cerevisiae under metal stress by two dimensional differential gel electrophoresis. AB - The defense mechanism by which cells combat metal stress remains poorly understood. By utilizing a newly developed technique - the differential gel electrophoresis (DIGE) - we evaluated the biological alterations of metal stress on Saccharomyces cerevisiae at its translational level. By simultaneously comparing the differential expression profiles of thousands of proteins as results of 15 different metal treatments, we were able to closely examine the response of a large number of proteins within the yeast proteome towards individual metals, as well as the response of the same proteins towards different metals. This, to our knowledge, is the first case which demonstrates the potential of DIGE as a high-throughput tool for large-scale proteome analysis. From our studies, where yeast cells were exhaustively treated with exogenous metals, 20-30% of all proteins detected showed statistically significant changes. According to different effects (up-/downregulation) of protein expression levels observed, we were able to tentatively divide the 15 metals into three groups. By mass spectrometric analysis, more than 50 protein spots were positively identified, both quantitatively and qualitatively. One of the proteins was identified to be Cu/Zn superoxide dismutase (SOD1), and its expression levels as a result of 15 different metal treatments was further examined in greater details. Significant changes in SOD1 expression were observed throughout all 15 DIGE gels. PMID- 12731035 TI - Reversed-phase high-performance liquid chromatography prefractionation prior to two-dimensional difference gel electrophoresis and mass spectrometry identifies new differentially expressed proteins between striate cortex of kitten and adult cat. AB - Two-dimensional difference gel electrophoresis (2-D DIGE), in combination with mass spectrometry, is a highly effective method for the rapid and reproducible detection of differentially expressed proteins. This approach, however, has the unfortunate drawback that it preferentially displays rather abundantly expressed proteins. Nevertheless, comparison of the protein expression levels of the striate cortex of adult cats and 30-day-old kittens, resulted in the identification of several proteins related to postnatal brain development and possibly age-dependent plasticity as well (Van den Bergh et al., J. Neurochem. 2003, in press). The goal of the present study was the selective enrichment and identification of less abundant proteins within the same paradigm. Hereto, we performed a reversed-phase chromatography prefractionation of our tissue lysate to separate the proteins in four fractions based on their hydrophobicity prior to 2-D DIGE analysis. This approach not only confirmed the differential expression levels of a number of proteins from the previous study, but also identified three additional proteins preferentially expressed in kitten visual cortex and five additional proteins with higher expression levels in adult cat visual cortex. These spots were not visible on the total tissue lysate protein maps, indicating that the high-performance liquid chromatography (HPLC) prefractionation enabled us to visualize additional, less abundant proteins. PMID- 12731039 TI - Stable expression of MHC class I heavy chain/HLA-DO complexes at the plasma membrane. AB - Major histocompatibility complex (MHC) class I and II molecules present antigenic fragments to the immune system. MHC-like chaperones, like HLA-DM, HLA-DO and tapasin support peptide loading. HLA class I heavy chains require association with beta 2-microglobulin and peptide for endoplasmic reticulum (ER) exit. Likewise, HLA-DO is only able to leave the ER by association to DM. Here we show that HLA-DO and free MHC class I heavy chains associate into a stable complex early during biosynthesis and are expressed at the plasma membrane as a complex. These DO/heavy chain complexes are found on DO-transfected cells and on low amounts on human B cells. PMID- 12731038 TI - SAP: a molecular switch regulating the immune response through a unique signaling mechanism. PMID- 12731040 TI - CD2 mediates activation of the IFN-gamma intronic STAT binding region in mucosal T cells. AB - The pathways leading to activation of mucosal lamina propria (LP) T cells differ from those of peripheral T cells. LP T cells exhibit enhanced IFN-gamma secretion when activated through the CD2 pathway. This study demonstrates CD2 signaling is followed by activation of STAT proteins in both peripheral blood mononuclear cells (PBMC) and lamina propria mononuclear cells (LPMC), although, distinct differences exist in regulation of IFN-gamma promoter gene expression. Both PBMC and LPMC exhibit enhanced secretion and transactivation of the -2.7 kb IFN-gamma promoter region following CD2 signaling, but the IFN-gamma STAT-binding region (within the first intron) serves as an orientation-independent enhancer of promoter activity only in LPMC. Mutation of the STAT site impairs enhancer activity. In LPMC, but not PBMC, CD2 mediates binding of STAT1 and STAT4 to the IFN-gamma intronic element. Unstimulated LMPC exhibit low levels of phosphotyrosine-STAT4 and STAT1 and phosphoserine-STAT1, which increase substantially following CD2 activation. In PBMC, CD2-mediated phosphorylation is primarily restricted to enhanced levels of phosphotyrosine-STAT1. Thus, these results indicate that both common as well as unique molecular mechanisms are involved in CD2 signaling and activation of the STAT pathway in LP T cells which are critical for regulation of IFN-gamma expression in the gut. PMID- 12731041 TI - Pax-5 is a key regulator of the B cell-restricted expression of the CD23a isoform. AB - Human CD23 (the low affinity IgE receptor) is a B cell differentiation marker involved in inflammatory responses. Two isoforms (CD23a and CD23b) are known, which differ only in their cytoplasmic domain. Whereas CD23b expression is specifically induced by IL-4 on B cells and cells of the myeloid lineage, CD23a expression is restricted to B cells. Each isoform is regulated by its own promoter. Pax-5 is a B-cell-restricted transcription factor with an essential role in early and late B cell development. Analyses of the CD23a promoter revealed a Pax-5-binding site, which can compete a high affinity Pax-5-binding site or directly bind Pax-5 protein in electrophoretic mobility shift assays. Introducing mutations into this site abrogates the binding. Expression of Pax-5 in 293 cells resulted in a seven- to tenfold activation of a CD23a core promoter construct. Most importantly, ectopic expression of Pax-5 in the monocytic cell line U-937, which regularly expresses only the CD23b isoform, led to CD23a expression after stimulation with IL-4 and PMA. Our results suggest that Pax-5 is a key regulator of the B-cell-restricted expression of the CD23a isoform. PMID- 12731042 TI - Effective and selective immune surveillance of the brain by MHC class I restricted cytotoxic T lymphocytes. AB - Cytotoxic CD8(+) T cells are abundantly present in human virus-induced or putative autoimmune diseases of the central nervous system (CNS). Their direct role in the induction of inflammatory brain damage is, however, poorly understood. We have studied CD8(+) T cell-mediated brain inflammation by transferring MHC class I-restricted hemagglutinin (HA)-reactive T cells from a TCR transgenic mouse line into transgenic mice, which express HA in astrocytes. We show that activated CD8(+) T cells alone can induce monophasic brain inflammation in immunocompetent recipient animals. Similar to previous studies, involving transfer of CD4(+) cells, brain inflammation peaks after 5-7 days and then declines. The pathology of brain inflammation, however, differs fundamentally from that induced by CD4(+) cells. The inflammatory reaction is dominated by T cells and activated microglia in the virtual absence of hematogenous macrophages. This is associated with exquisitely specific destruction of antigen-containing astrocytes in the absence of any bystander damage of myelin, oligodendrocytes or neurons. Furthermore, in contrast to CD4(+) T cells, some CD8(+) cells accumulate in the brain and activate microglia in recipient animals, even in the absence of the specific antigen in the CNS. These data indicate that CD8(+) T cells are prime candidates for immune surveillance of the CNS. PMID- 12731043 TI - Tumor rejection by gene transfer of the MHC class II transactivator in murine mammary adenocarcinoma cells. AB - The murine mammary adenocarcinoma cell line TS/A is a highly malignant MHC class II-negative tumor. We show that transfection of TS/A cells with the MHC class II transactivator CIITA renders them MHC class II-positive and highly immunogenic in vivo. These cells were fully rejected by 51% of syngeneic recipients and had a significantly lower growth rate in the remaining 49% of animals. This directly correlated to the amount of MHC class II molecules expressed in the transfected tumor. Tumor rejecting animals were protected against rechallenge with the parental TS/A tumor. The rejection required CD4(+) and CD8(+) T cells. CD4(+) T cells were fundamental in the priming phase of the antitumor response. CTL specific for a peptide of the envelope gp70 of an endogenous ecotropic retrovirus were identified and explained the specificity of the effector mechanism of rejection against the TS/A and the antigenically related C26 carcinoma cells but not against the unrelated gp70-negative syngeneic fibrosarcoma F1F cells. This is the first example of successful tumor vaccination by genetic transfer of CIITA. These results open the way to a possible use of CIITA for increasing both the inducing and the effector phase of the antitumor immune response. PMID- 12731044 TI - A role for antibodies in the generation of memory antifungal immunity. AB - Protective immunity to Candida albicans and Aspergillus fumigatus is mediated by antigen-specific Th1 cells. To define the role of B cells and antibodies in the generation of antifungal immune resistance, B cell-deficient (mu MT) mice were assessed for immune resistance to primary and secondary infections with both fungi. The results showed that, although passive administration of antibodies increased the fungal clearance, the innate and Th1-mediated resistance to the primary and secondary infections were both heightened in mu MT mice with candidiasis and aspergillosis. However, although capable of efficiently restricting the fungal growth, mu MT mice did not survive the re-infection with C. albicans, and this was concurrent with the failure to generate IL-10-producing dendritic cells and regulatory CD4(+)CD25(+) T cells. Antifungal opsonizing antibodies restored IL-10 production by dendritic cells from mu MT mice, a finding suggesting that the availability of opsonizing antibodies may condition the nature of the dendritic cell interaction with fungi, possibly impacting on the development of long-lasting antifungal immunity. PMID- 12731046 TI - T cell adhesion lowers the threshold for antigen detection. AB - Antigen detection and initiation of TCR signaling only occur, under physiological conditions, when T cells are adherent, and not in suspension. We show here that T cell adhesion causes an increase in the Ca(2+) content of intracellular stores and of the amount of phosphatidylinositol 4,5-bisphosphate in the plasma membrane, and enhances TCR-induced Ca(2+) signaling. This priming can be observed in freshly isolated T cells, in activated T cells, and in some T cell lines. Stimulation of T cells by specific monomeric MHC-peptide complexes only triggers Ca(2+) responses after T cell adhesion. This solves a controversial issue concerning the minimum valency of activatory TCR ligands. Adhesion-induced T cell priming not only occurs upon binding to artificial substrates such as immobilized ligands, but also upon interaction with dendritic cells. Thus, this phenomenon is likely to contribute to the high sensitivity of antigen detection by T cells in secondary lymphoid organs. PMID- 12731045 TI - IL-10 and TGF-beta cooperate in the regulatory T cell response to mucosal allergens in normal immunity and specific immunotherapy. AB - The regulation of normal and allergic immune responses to airborne allergens in the mucosa is still poorly understood, and the mechanism of specific immunotherapy (SIT) in normalizing the allergic response to such allergens is currently not clear. Accordingly, we have investigated the immunoregulatory mechanism of both normal and allergic responses to the major house-dust mite (HDM) and birch pollen allergens--Dermatophagoides pteroynyssinus (Der p)1 and Bet v 1, respectively--as well as the immunologic basis of SIT to HDM in rhinitis and asthma patients. In normal immunity to HDM and birch pollen, an allergen specific peripheral T cell suppression to Der p 1 and Bet v 1 was observed. The deviated immune response was characterized by suppressed proliferative T cell and Th1 (IFN-gamma) and Th2 (IL-5, IL-13) cytokine responses, and increased IL-10 and TGF-beta secretion by allergen-specific T cells. Neutralization of cytokine activity showed that T cell suppression was induced by IL-10 and TGF-beta during SIT and in normal immunity to the mucosal allergens. In addition, SIT induced an antigen-specific suppressive activity in CD4(+) CD25(+) T cells of allergic individuals. Together, these results demonstrate a deviation towards a regulatory/suppressor T cell response during SIT and in normal immunity as a key event for the healthy immune response to mucosal antigens. PMID- 12731047 TI - Organ-specific and stage-dependent control of Leishmania major infection by inducible nitric oxide synthase and phagocyte NADPH oxidase. AB - In the Leishmania major mouse model of cutaneous leishmaniasis inducible nitric oxide synthase (iNOS) is crucial for the killing of the parasite in the skin and draining lymph node. However, the effector mechanism operating against L. major in the spleen is unknown. As reactive oxygen intermediates might play a role, we analyzed macrophages and mice lacking the gp91phox subunit of the phagocyte NADPH oxidase (phox) for their ability to combat an infection with L. major. Macrophages from wild-type and gp91phox(-/-) mice had an equal capacity to kill L. major after activation by cytokines. Unlike iNOS, the activity of phox was dispensable for the resolution of the acute skin lesions and exerted only a limited effect on the containment of the parasites in the draining lymph node, but was essential for the clearance of L. major in the spleen. During the chronic phase of infection, parasites persisted at high levels in gp91phox(-/-) mice, and cutaneous lesions re-emerged in approximately 60% of these mice. gp91phox deficiency did not impair the expression of iNOS or the production of TNF and IFN gamma. These results demonstrate that iNOS and phox are both required for the control of L. major in vivo and display unexpected organ- and stage-specific anti leishmanial effects. PMID- 12731049 TI - Idiotype-specific CD4+CD25+ T suppressor cells prevent, by limiting antibody diversity, the occurrence of anti-dextran antibodies crossreacting with histone H3. AB - CD25(+) suppressor T cells regulate the immune response against the type-2 "thymus independent" bacterial polysaccharide antigen alpha(1-->3)dextran (Dex) in BALB/c mice. These T cells, represented by the clone 178-4 Ts, restrict the Dex-specific IgG antibody repertoire such that the J558 idiotype dominates. Antibodies with other structures in the heavy-chain variable region (V(H) region), predominantly within the CDR3 domain, occur when the T cell control fails. This increase of antibody diversity caused by a lack of CD25(+) Ts cells, e.g. in nude mice, does not result in the appearance of antibodies with enhanced affinity to the antigen Dex, but often leads to a crossreactivity with autologous proteins. Twenty-two out of sixty Dex-specific hybridomas from nude mice, but no hybridomas from euthymic mice, crossreact with a nuclear protein, as tested by ELISA. This nuclear protein was identified as histone H3. Ten of the sixty hybridomas from nude mice were sequenced and show V(H) sequences that deviate from the original J558 sequence. Three of these ten hybridomas crossreact with the histone H3. Adoptive transfer of CD25(+) Ts cells to nude mice leads to a marked increase of antibodies carrying the original J558 idiotype within the IgG pool after immunization with Dex. Our data demonstrate a CD25(+) Ts cell-mediated restriction of V(H) usage, which prevents the appearance of crossreactive autoantibodies. PMID- 12731048 TI - Selective cross-talk among natural cytotoxicity receptors in human natural killer cells. AB - The cytolytic activity of human natural killer cells is induced by several triggering cell surface receptors upon interaction with specific cellular ligands. These receptors include NKp46, NKp30 and NKp44, collectively termed natural cytotoxicity receptors (NCR). Co-operation among NCR has been shown to occur for optimal recognition and killing of most tumor target cells. In this study, we show that the mAb-mediated engagement and clustering of one or another NCR results in the activation of an identical set of tyrosine kinases. These kinases are included in the signaling cascade leading to tyrosine phosphorylation of different receptor-associated signal transducing molecules i.e. CD3 zeta (associated with NKp46 and NKp30) and KARAP/DAP12 (associated with NKp44). In line with the notion that the engagement of inhibitory receptors prevents NCR mediated responses, we show that the engagement of CD94/NKG2A virtually abrogates the tyrosine phosphorylation of the NCR-associated signaling molecules, i.e. it acts at the very early steps of the signaling cascade. Importantly, the engagement of a single NCR resulted in the activation of the signaling cascades associated with the other NCR. This "cross-talk" is confined to NKp46, NKp30 and NKp44 since neither CD16-nor KIR2DS4-associated signaling polypeptides were phosphorylated following the NCR engagement. These results suggest that a functional cross-talk specifically occurs among different NCR, possibly resulting in the amplification of the activating signals. PMID- 12731050 TI - Major histocompatibility complex class II-restricted presentation of a cytosolic antigen by autophagy. AB - Biochemical and functional studies have demonstrated major histocompatibility complex (MHC) class II-restricted presentation of peptides derived from cytosolic proteins, but the underlying processing and presentation pathways have remained elusive. Here we show that endogenous presentation of an epitope derived from the cytosolic protein neomycin phosphotransferase II (NeoR) on MHC class II is mediated by autophagy. This presentation pathway involves the sequestration of NeoR into autophagosomes, and subsequent delivery into the lytic compartment. These results identify endosomes/lysosomes as the processing compartment for cytosolic antigens and furthermore link endogenous antigen presentation on MHC class II with the process of cellular protein turnover by autophagy. PMID- 12731051 TI - Intracerebroventricular injection of the terminal complement complex causes inflammatory reaction in the rat brain. AB - To investigate the effect of the terminal complement complex (TCC) on the central nervous system, we injected both the cytolytically active and the inactive complexes into the lateral ventricle of rats. Both complexes promoted accumulation of leukocytes into the cerebrospinal fluid at 4-6 h post-injection. The cells recovered at this time were mostly polymorphonuclear leukocytes (PMN) that were partially replaced by mononuclear cells at 12 h. A direct contribution of the complexes to the in-vivo migration of leukocytes was ruled out by their inability to be chemotactic for rat PMN. Contaminating C5a is unlikely to be responsible for the effect of TCC because it failed to mobilize leukocytes when injected into the lateral ventricle. Histological analysis of rat brains 6 hours after injection of TCC revealed marked leukocyte infiltration of the choroid plexus, increased expression of intercellular adhesion molecule-1 and egression of leukocytes out of the meningeal vessels. The cerebrospinal fluid of rats treated with TCC exhibited chemotactic activity for rat PMN and increased levels of growth related oncogene/cytokine-induced neutrophil chemoattractant-1 and monocyte chemoattractant protein-1 preceding the accumulation of leukocytes. Elevated concentration of IL-1 beta was also found in the cerebrospinal fluid and in periventricular areas of rats treated with TCC. PMID- 12731052 TI - Schistosome N-glycans containing core alpha 3-fucose and core beta 2-xylose epitopes are strong inducers of Th2 responses in mice. AB - During murine schistosomiasis, egg-derived glycoconjugates play a key role in skewing the immune response towards a Th2 phenotype. Among the candidates responsible for this effect, complex-type N-glycans containing the core alpha 3 fucose and core beta 2-xylose determinants, two glycan epitopes found in some invertebrate- and plant-derived allergens, may be important. Here, we show that core alpha 3-fucose and core beta 2-xylose determinants are expressed in the different developmental stages of Schistosoma mansoni, particularly in the excretory-secretory systems of schistosomula and adult worms and in eggs deposited in the liver. Glycosyltransferase assays confirmed the presence of core alpha 3-fucosyltransferase and core beta 2-xylosyltransferase activities in egg extracts. Using a model of immunization with pulsed dendritic cells, we show that egg-derived glycoproteins containing the core alpha 3-fucose and core beta 2 xylose determinants generate a strong Th2-biased cellular response in mice and that the glycan moieties of this extract are important in this effect. During murine infection, these complex-type N-glycans induce a glycan-specific Th2 cellular response and elicit T-dependent anti-core alpha 3-fucose and anti-core beta 2-xylose IgG1 (a Th2-associated isotype), but not IgG2b (a Th1-associated isotype) Ab. Taken together, our results point out the importance of core fucosylated/xylosylated N-glycans in the Th2 immune response during murine schistosomiasis. PMID- 12731053 TI - Decreasing the threshold for thymocyte activation biases CD4+ T cells toward a regulatory (CD4+CD25+) lineage. AB - Thymus-derived CD4(+)CD25(+) regulatory T (T(r)) cells play a critical role in suppressing aberrant responses to self in vivo. The factors that influence a CD4(+) T cell's decision to commit to an immunoregulatory T(r) cell lineage are currently unknown. In the present study, we found that in mice, abundantly expressing a few or one peptide(s) bound to MHC class II molecules, a large portion of conventional CD4(+) T cells could be biased towards the commitment to a T(r) lineage by reducing the threshold required for thymocyte activation. This occurred in the presence of either an antisense glucocorticoid receptor transgene or a pharmacological inhibitor of glucocorticoid synthesis. These results demonstrate a novel in vivo pathway for the generation of T(r) cells, and raise the possibility that therapeutic enhancement of the T(r) cell repertoire through pharmacological manipulation of TCR signaling thresholds may provide a feasible means of ameliorating autoimmunity. PMID- 12731054 TI - Visualization of peptide presentation following oral application of antigen in normal and Peyer's patches-deficient mice. AB - Orally applied antigens generate systemic unresponsiveness by induction of anergy and deletion of specific T cells at high antigen doses, and induction of regulatory T cells at low doses of antigen. These different immune reactions have been attributed to different types of antigen-presenting cells (APC) and/or different secondary lymphoid organs participating in the induction of the immune response. We used high-sensitivity immunofluorescence to directly identify for the first time the cells presenting orally applied antigen in vivo. At low peptide doses (<1 mg) peptide presentation was exclusively detected on dendritic cells (DC) of the Peyer's patches (PP) and mesenteric lymph nodes (mLN). At high doses (>1 mg) peptides were presented systemically and by all types of APC but presentation was still maximal on DC of the PP (up to 65%). Nevertheless, at limiting antigen doses T cell activation in the gut-associated lymphoid tissue occurs preferentially in the mLN but not in PP. PP-deficient mice have the same frequencies of peptide-presenting cells in mLN, peripheral lymph nodes and spleen and activation of naive T cells in vivo is not affected. Therefore, PP are not critical for antigen presentation as well as for T cell activation in response to orally applied soluble antigens. PMID- 12731055 TI - Proteoglycans are potent modulators of the biological responses of eosinophils to chemokines. AB - Chemokines play critical roles in governing the recruitment and activation of eosinophils at sites of allergic inflammation, particularly the asthmatic lung. However, we know little of how chemokine function is regulated post translationally. Proteoglycans, consisting of a core protein and glycosaminoglycan (GAG) side chains, are cell surface molecules and components of the extracellular matrix that are able to bind chemokines, whilst heparin is a GAG with therapeutic value in asthma. We examined whether soluble GAG could alter the actions of chemokines in assays of eosinophil activation. Heparin inhibited intracellular calcium flux, respiratory burst and chemotactic responses of eosinophils to CCL11, but not to the chemoattractant C5a, and inhibited binding of CCL11 to CCR3. Heparin also inhibited eosinophil stimulation by CCL11, CCL24, CCL7, CCL13 and CCL5 to differing degrees, which broadly correlated with their relative affinities for heparin. Heparan sulfate and dermatan sulfate, but not chondroitin sulfate, also inhibited the actions of CCL11 and CCL13 in assays of cellular shape change and chemotaxis. Following treatment with the sulfation inhibitor chlorate or proteoglycanases, no inhibition of CCL11-induced activity was observed using either eosinophils or a CCR3-expressing transfectant cell line. This suggests that cell surface proteoglycans are not necessary for signaling via CCR3. However, the GAG context in which chemokines are expressed is likely to represent an important level of regulation of allergic inflammation. PMID- 12731056 TI - Tumor-associated fibroblasts recruit blood monocytes into tumor tissue. AB - Tumor-associated fibroblasts (TAF) and tumor-associated macrophages are the main stromal components in desmoplastic breast tumors. These host cell types were extensively studied individually with regard to tumor development and progression but little is known about their reciprocal interactions. To elucidate the role of TAF in the recruitment of monocytes (MO) we designed a 3D co-culture system of multicellular fibroblast spheroids of different origin, co-cultured with MO suspensions from healthy donors. Spheroids of tumor-derived but not of normal fibroblasts were extensively infiltrated by MO. A linear correlation between number of infiltrated MO and number of MO applied per spheroid was shown, indicating a distinct migratory MO subpopulation (approximately 15%) within the peripheral blood MO pool. Our data imply that MO migration into fibroblastic tumor areas may partially result from high expression of CCL2 (monocyte chemotactic protein-1), which was regulated by endogenous IL-6 as shown by neutralization experiments. The effect of CCL2 on MO migration was inhibited by CCL2 neutralizing antibody in tumor-derived fibroblast conditioned media in a Boyden chamber migration assay but not in spheroid culture. While this phenomenon needs further evaluation, our data clearly support the concept of fibroblasts as "sentinel cells" relevant for tumor progression. PMID- 12731057 TI - Immunogenicity and protective efficacy of Plasmodium falciparum liver-stage Ag-3 in Aotus lemurinus griseimembra monkeys. AB - Three recombinant proteins spanning the Plasmodium falciparum liver-stage Ag-3 (LSA-3) were used to immunize Aotus monkeys. The proteins were delivered subcutaneously without adjuvant, adsorbed onto polystyrene 0.5 microm particles at a concentration of 2 microg per immunization. Control animals received glutathione-S-transferase formulated similarly. Animals were challenged as late as 5 months after the last immunization, by intravenous inoculation of 100,000 P. falciparum sporozoites of a strain heterologous to the one from which the immunogens were derived. Sterile protection was achieved in three of the five immunized monkeys but in none of four controls. Antibodies were at low titer, but reacted with the native parasite protein and were boosted by parasite challenge. Ag-specific IFN-gamma secretion was detectable in all LSA-3-immunized animals in response to the LSA-3-derived Ag. The protection was apparently associated with high levels of IFN-gamma production in response to in vitro recall Ag. These results lend support to the vaccine potential of LSA-3 indicated by previous results obtained in chimpanzees, as well as the value of yet another Ag-delivery system. They also support the value of the Aotus model for the pre-clinical development of pre-erythrocytic-stage vaccines. PMID- 12731058 TI - Chemokines integrate JAK/STAT and G-protein pathways during chemotaxis and calcium flux responses. AB - The JAK/STAT (Janus kinase / signaling transducer and activator of transcription) signaling pathway is implicated in converting stationary epithelial cells to migratory cells. In mammals, migratory responses are activated by chemoattractant proteins, including chemokines. We found that by binding to seven-transmembrane G protein-coupled receptors, chemokines activate the JAK/STAT pathway to trigger chemotactic responses. We show that chemokine-mediated JAK/STAT activation is critical for G-protein induction and for phospholipase C-beta dependent Ca(2+) flux; in addition, pharmacological inhibition of JAK or mutation of the JAK kinase domain causes defects in both responses. Furthermore, G alpha(i) association with the receptor is dependent on JAK activation, and the chemokine mediated Ca(2+) flux that requires phospholipase C-beta activity takes place downstream of JAK kinases. The chemokines thus employ a mechanism that links heterologous signaling pathways--G proteins and tyrosine kinases--in a network that may be essential for mediating their pleiotropic responses. PMID- 12731059 TI - A recombinant bispecific single-chain antibody induces targeted, supra-agonistic CD28-stimulation and tumor cell killing. AB - Endowing tumor cells with costimulatory signals for T cell activation has emerged as a promising strategy for tumor immunotherapy. Costimulatory molecules were either transfected into tumor cells to generate vaccines or were fused, e.g. to antibodies against tumor-associated antigens, to achieve targeted T cell costimulation in vivo. Here we report the production and purification of rM28, a recombinant bispecific single-chain antibody directed to a melanoma-associated proteoglycan and to the costimulatory CD28 molecule on human T cells. We found that a dimer of the recombinant molecule, bound to tumor target cells, induced pronounced T cell activation in peripheral blood mononuclear cell preparations without additional TCR/CD3 stimulation being required. The lytic activity generated after 3 days of stimulation effectively prevented tumor cell growth. However, it was unspecific and predominantly mediated by non T cells. Our findings demonstrate that presentation of a CD28 antibody within a suitable recombinant, bispecific format may result in a "targeted supra-agonistic stimulation" of the CD28 molecule, which leads to effective tumor cell killing after induction of unspecifically lytic cells. PMID- 12731060 TI - TNF-alpha impairs peripheral tolerance towards beta-cells, and local costimulation by B7.1 enhances the effector function of diabetogenic T cells. AB - Maintenance of peripheral tolerance and inactivation of autoreactive T cells is based on a delicate balance between pro-inflammatory and protective cytokines that is poorly understood. We have here addressed how the local expression of the inflammatory cytokine TNF-alpha can impair peripheral tolerance and lead to autoreactivity. After transplantation of pancreata that are immunogenic due to beta-cell expression of B7.1 and TNF-alpha, into thymectomized and euthymic syngeneic mice, we found that only euthymic mice rejected the grafts. This result suggests that under normal circumstances autoreactive T cells are functionally inactivated, and initiation of an autoreactive response requires de-novo generation of T cells. By contrast, thymectomized mice expressing TNF-alpha on the endogenous islets rejected the grafts, showing that expression of TNF-alpha prevents functional silencing of the autoreactive T cells. Thus, this study provides a mechanism by which TNF-alpha and possibly chronic inflammatory responses may promote autoimmune diseases. Furthermore, we have investigated whether B7.1 can enhance T cell responses of already activated T cells leading to islet rejection. By transplantation of wild-type and B7.1-expressing islets into overtly diabetic mice we found that only the wild-type islets could restore normoglycemia, suggesting that costimulation by B7.1 is required in the expansion or effector phase of the response. PMID- 12731062 TI - Lymphocyte trafficking through blood and lymphatic vessels: more than just selectins, chemokines and integrins. PMID- 12731061 TI - ICAM-1 isoforms: specific activity and sensitivity to cleavage by leukocyte elastase and cathepsin G. AB - The extracellular moiety of ICAM-1 consists of five Ig-like domains, the first and third domains mediating adhesion to integrin ligands. The ICAM-1 gene, however, gives rise to the expression of five alternative splice variants containing two, three, or four Ig-like domains. In this work, we have investigated whether the rearrangement of the architecture of ICAM-1 affects its structural properties and function. We showed that, in contrast to the common form, all alternative isoforms of ICAM-1 were susceptible to cleavage by leukocyte elastase and cathepsin G. We found that the length of an isoform did not influence the susceptibility to proteolysis. The molecular diversity provided by the skipping of entire Ig domains and the level of expression on the APC, however, significantly influenced their ability to potentiate the proliferation of T cells. Finally, we found that the expression of minor ICAM-1 isoforms encoding the third Ig-like domains was sufficient to sustain neutrophil infiltration in the liver and confer exon-5-targeted ICAM-1-deficient mice susceptibility to LPS-induced septic shock. These findings not only demonstrate that ICAM-1 isoforms are fully functional, but support the concept that alternative RNA splicing in the Ig superfamily may fulfill distinct roles during the development of the immune response. PMID- 12731063 TI - Inactivation of the EGF-TM7 receptor EMR4 after the Pan-Homo divergence. AB - We here report on the identification of a novel human EGF-TM7 receptor, designated EMR4. Like most EGF-TM7 receptor genes, EMR4 is localized on the short arm of chromosome 19, in close proximity to EMR1. Remarkably, due to a one nucleotide deletion in exon 8, translation of human EMR4 would result in a truncated 232-amino acid protein lacking the entire seven-span transmembrane region. This deletion is not present in nonhuman primates, including chimpanzees, suggesting that EMR4 became nonfunctional only after human speciation, about five million years ago. Thus, EMR4 surprisingly accounts for a genetic difference between humans and primates related to immunity. PMID- 12731064 TI - TGF-beta1 modulates Fas (APO-1/CD95)-mediated apoptosis of human pre-B cell lines. AB - We have previously shown that Fas-induced apoptosis is markedly enhanced by IL-7 in human pre-B but not pro-B cell lines. In addition, pre-B cell receptor (pre BCR) ligation significantly potentiates the IL-7 effects on Fas-triggered pre-B cell death. We show herein that transforming growth factor (TGF)-beta 1 sharply reduces Fas-induced death rate of pre-B but not pro-B cells. TGF-beta 1 causes inhibition of Fas-mediated disruption of mitochondrial transmembrane potential and cleavage of caspase 8, Bid and caspase 3. Bcl2 expression is markedly increased in TGF-beta 1-treated pre-B cells, whereas cellular FLICE-like inhibitory protein long (c-FLIPL), Bcl-XL, Bax, and Bad expression remains unchanged. TGF-beta 1 causes a selective growth arrest of pre-B cells in G0/G1 phase of the cell cycle and induces a partial down-modulation of both Fas and pre BCR expression. All TGF-beta 1-mediated effects, but Bcl2 up-regulation, can be reproduced by the LY294002 phosphatidylinositol 3-kinase (PI3K)/Akt inhibitor but not by inhibitors of the MAPK/ERK (MEK) and Janus kinase (Jak)/STAT pathways, which promote cell death. Akt phosphorylation is strongly inhibited by TGF-beta1 in pre-B but not pro-B cells and is not modified by Fas engagement. Altogether, our findings suggest that TGF-beta1 prevents Fas-induced apoptosis of pre-B lines by inhibiting PI3K pathway and by enhancing expression of Bcl2. They also suggest that the PI3K/Akt pathway is involved in the control of Fas and pre-BCR expression, a checkpoint in B cell development. PMID- 12731065 TI - Immunostimulatory effects of plasmid DNA and synthetic oligodeoxynucleotides. AB - Nucleic acid immunization is a new vaccination technology. DNA vaccines do not only carry the genetic information for the antigen of interest but also deliver an adjuvant effect due to the presence of immunostimulatory sequences within the plasmid backbone. It is generally assumed that the adjuvant properties of plasmid DNA are equal to those described for oligodeoxynucleotides (ODN) containing immunostimulatory CpG motifs. To challenge this hypothesis we have carried out a series of experiments comparing the ability of single- and double-stranded ODN containing CpG motifs to induce the activation of mouse spleen cells. Moreover, we compared the immunostimulatory properties of plasmids that were modified by the addition of two to four CpG motifs. Our results establish that plasmid DNA express their adjuvanticity as either double or single strands, and no differences were observed between modified and unmodified plasmids. On the other hand, the strongest stimulatory ODN sequences lost their adjuvant properties when administered as double-strand DNA. Furthermore, the profile of cytokines induced on spleen cells by plasmid DNA and ODN is different. Strikingly, plasmid DNA induces a moderate synthesis of IL-6 and a strong synthesis of IFN-gamma, whereas stimulation with ODN showed an inverse profile with a higher increase in the synthesis of IL-6 but a moderate increase in IFN-gamma. Finally, in vivo studies were consistent with the results obtained in vitro. Mice immunized with modified or unmodified plasmids encoding the glycoprotein D of HSV showed similar levels of cellular and humoral immune responses. PMID- 12731066 TI - Redundant functions of TCF-1 and LEF-1 during T and NK cell development, but unique role of TCF-1 for Ly49 NK cell receptor acquisition. AB - Members of the TCF/LEF (T cell factor / lymphoid enhancer factor) family of DNA binding factors play important roles during embryogenesis, the establishment and/or maintenance of self-renewing tissues such as the immune system and for malignant transformation. Specifically, it has been shown that TCF-1 is required for T cell development. A role for LEF-1 became apparent when mice harbored two hypomorphic TCF-1 alleles and consequently expressed low levels of TCF-1. Here we show that NK cell development is similarly regulated by redundant functions of TCF-1 and LEF-1, whereby TCF-1 contributes significantly more to NK cell development than LEF-1. Despite this role for NK cell development, LEF-1 is not required for the establishment of a repertoire of MHC class I-specific Ly49 receptors on NK cells. The proper formation of this repertoire depends to a large extent on TCF-1. These findings suggest common and distinct functions of TCF-1 and LEF-1 during lymphocyte development. PMID- 12731067 TI - CD55/decay accelerating factor is part of the lipopolysaccharide-induced receptor complex. AB - Recently, we described an 80-kDa lipopolysaccharide (LPS)-binding membrane protein to be identical to CD55 [decay accelerating factor (DAF)]. Here, we demonstrate that CD55 is able to contribute to lipopolysaccharide (LPS) signaling. Transfection of Chinese hamster ovary (CHO) cells with human CD55 resulted in a translocation of NF-kappa B after stimulation with LPS as well as with free lipid A. In addition, interaction of lipid A and CD55 was shown by co immuno-precipitation of these molecules from CHO-CD55 cells after incubation with lipid A and anti-lipid A monoclonal antibody, as well as by fluorescence resonance energy transfer (FRET) analysis in human monocytes. The comparison of LPS-induced signaling pathways in CHO-CD55 and CHO-CD14 cells revealed that p38, JNK and ERK MAP kinases are activated upon LPS stimulation in both cell lines, and that the activation by LPS can be blocked at the level of Toll-like receptor 4. Finally, through FRET analysis we could demonstrate LPS-induced clustering of CD55 and CD11/CD18 in human monocytes. Our results imply a new functional role of CD55 as a member of a multimeric LPS receptor complex. PMID- 12731068 TI - Low-avidity recognition by CD4+ T cells directed to self-antigens. AB - Self-reactive T cells populate the peripheral immune system, and likely form the reservoir from which autoreactive cells are derived. We analyzed a panel of self and non-self peptides presented by HLA-DR4, a class II molecule associated with autoimmunity, by immunization of mice transgenic for HLA-DR4. Significant structural avidity for T cell recognition, as measured by MHC class II tetramer binding to CD4(+) T cells was only observed in mice immunized with the non-self antigens. T cell hybridomas were generated from mice immunized with the naturally processed self-peptide hGAD65 (552-572) and also from mice immunized with an influenza-derived non-self epitope (HA 306-318). T cells specific for the self peptide failed to bind tetramers and exhibited low functional avidity as measured by the peptide concentration required to reach half-maximum proliferation values. In contrast, T cells specific for the non-self HA (306-318) peptide exhibited high structural and functional avidity profiles. As recently described in studies of murine CD8(+) T cell function, the predominance of low avidity recognition of self-peptide epitopes may be a characteristic feature of CD4(+) T cells responding to autoantigens. PMID- 12731069 TI - Mice with neonatally induced inactivation of the vascular cell adhesion molecule 1 fail to control the parasite in Toxoplasma encephalitis. AB - Under various inflammatory conditions, cell adhesion molecules are up-regulated in the central nervous system (CNS) and may contribute to the recruitment of leukocytes to the brain. In the present study, the functional role of vascular cell adhesion molecule (VCAM)-1 in Toxoplasma encephalitis (TE) was addressed using VCAM(flox/flox MxCre) mice. Neonatal inactivation of the VCAM-1 gene resulted in a lack of induction of VCAM-1 on cerebral blood vessel endothelial cells, whereas the constitutive expression of VCAM-1 on choroid plexus epithelial cells and the ependyma was unaffected; in these animals, resistance to T. gondii was abolished, and VCAM(flox/flox MxCre) mice died of chronic TE caused by a failure to control parasites in the CNS. Although leukocyte recruitment to the CNS was unimpaired, the B cell response was significantly reduced as evidenced by reduced serum levels of anti-T. gondii-specific IgM and IgG antibodies. Furthermore, the frequency and activation state of intracerebral T. gondii specific T cells were decreased, and microglial activation was markedly reduced. Taken together, these data demonstrate the crucial requirement of VCAM-1-mediated immune reactions for the control of an intracerebral infectious pathogen, whereas other cell adhesion molecules can efficiently compensate for VCAM-1-mediated homing across cerebral blood vessels. PMID- 12731070 TI - Fibrinogen-CD11b/CD18 interaction activates the NF-kappa B pathway and delays apoptosis in human neutrophils. AB - The regulation of neutrophil half-life by members of the coagulation cascade is critical for the resolution of the inflammatory response. We have demonstrated that soluble fibrinogen (sFbg) delays human neutrophil (PMN) apoptosis through a mechanism that involves CD11b interactions, and phosphorylation of focal adhesion kinase (FAK) and extracellular signal-regulated kinase 1/2 (ERK1/2). Since NF kappa B is a key element in the regulation of apoptotic mechanisms in several immune cells, we investigated whether NF-kappa B is involved in the control of PMN survival by sFbg. We show that sFbg triggers inhibitor protein kappa B (I kappa B-alpha) degradation and NF-kappa B activation. Furthermore, pharmacological inhibition of NF-kappa B abrogates sFbg effects on apoptosis. In addition, specific inhibition of MAPK ERK1/2 significantly reduces NF-kappa B translocation by sFbg, suggesting a relationship between ERK1/2 and NF-kappa B activation. Similar results are obtained when granulocytic-differentiated HL-60 cells are treated with sFbg, making this model highly attractive for integrin induced gene expression studies. It can be concluded that NF-kappa B participates in the prevention of apoptosis induced by sFbg with the participation of MAPK ERK1/2. These results shed light on the molecular mechanisms that control human granulocyte apoptosis, and suggest that NF-kappa B regulation may be of benefit for the resolution of the inflammatory response. PMID- 12731071 TI - Schistosoma mansoni antigens modulate the activity of the innate immune response and prevent onset of type 1 diabetes. AB - Infection with Schistosoma mansoni (S. mansoni) or exposure to eggs from this helminth inhibits the development of type 1 diabetes in NOD mice. In this study we show that soluble extracts of S. mansoni worm or egg completely prevent onset of type 1 diabetes in these mice but only if injection is started at 4 weeks of age. T cells from diabetes-protected mice make IL-10 in recall responses to parasite antigens. These cells are furthermore impaired in their ability to transfer diabetes to NOD-SCID recipients. Bone marrow dendritic cells derived from NOD mice are found to make more IL-10 and less IL-12 following culture with S. mansoni soluble egg antigens in conjunction with lipopolysaccharides. NOD mice are deficient in NKT cells. Soluble worm and egg antigens increase the numbers of V alpha 14i NKT cells in NOD mice. These effects of schistosome antigens on the innate immune system provide a mechanism for their ability to prevent type 1 diabetes in NOD mice. PMID- 12731074 TI - AAA award winners. PMID- 12731072 TI - Comparison of Fc epsilon RI- and Fc gamma RI-mediated degranulation and TNF-alpha synthesis in human mast cells: selective utilization of phosphatidylinositol-3 kinase for Fc gamma RI-induced degranulation. AB - We have demonstrated that CD34(+) IFN-gamma-treated human mast cells (HuMC) express functional Fc gamma RI and that aggregation of these receptors leads to mediator release. As the signaling pathways linking Fc gamma RI aggregation to mediator release are unknown, we examined Fc gamma RI-dependent activation of specific signal transduction molecules and determined the relative involvement of these events in HuMC degranulation and TNF-alpha production following both Fc gamma RI and Fc epsilon RI aggregation. Fc gamma RI aggregation resulted in the phosphorylation/activation of src kinases and p72(syk) and subsequent tyrosine phosphorylation of multiple substrates. Inhibitor studies revealed that these responses were required for degranulation and TNF-alpha synthesis. Both Fc gamma RI and Fc epsilon RI aggregation also activated the MAP kinases ERK 1/2, JNK and p38 and this was necessary for TNF-alpha synthesis, but not degranulation for both receptors. Thus, signaling events in HuMC following aggregation of Fc gamma RI were generally similar to those observed following Fc epsilon RI aggregation. The one exception was that, although phosphatidylinositol-3-kinase was activated after both Fc epsilon RI and Fc gamma RI aggregation, only the Fc gamma RI appeared to require this molecule for degranulation. PMID- 12731075 TI - Using a modified standard microscope to generate virtual slides. AB - A standard microscope was reconfigured as a virtual slide generator by adding a Prior Scientific H101 robotic stage with H29 controller and 0.1 microm linear scales and a Hitachi HV-C20 3CCD camera. Media Cybernetics Image Pro Plus version 4 (IP4) software controlled stage movement in the X-, Y-, and Z-axis, whereas a Media Cybernetics Pro-Series Capture Kit captured images at 640 x 480 pixels. Stage calibration, scanning algorithms, storage requirements, and viewing modes were standardized. IP4 was used to montage the captured images into a large virtual slide image that was subsequently saved in TIF or JPEG format. Virtual slides were viewed at the workstation using the IP4 viewer as well as Adobe Photoshop and Kodak Imaging. MGI Zoom Server delivered the virtual slides to the Internet, and MicroBrightField's Neuroinformatica viewing software provided a browser-based virtual microscope interface together with labeling tools for annotating virtual slides. The images were served from a Windows 2000 platform with 2 GB RAM, 500 GB of disk storage, and a 1.0 GHz P4 processor. To conserve disk space on the image server, TIF files were converted to the FlashPix (FPX) file format using a compression ratio of 10:1. By using 4x, 10x, 20x, and 40x objectives, very large gigapixel images of tissue whole-mounts and tissue arrays with high quality and morphologic detail are now being generated for teaching, publication, research, and morphometric analysis. Technical details and a demonstration of our system can be found on the Web at http://virtualmicroscope.osu.edu. PMID- 12731078 TI - When gold is not noble: catalysis by nanoparticles. AB - Bulk gold is chemically inert and is generally regarded as a poor catalyst. However, when gold is in very small particles with diameters below 10 nm and is deposited on metal oxides or activated carbon, it becomes surprisingly active, especially at low temperatures, for many reactions such as CO oxidation and propylene epoxidation. The catalytic performance of Au is defined by three major factors: contact structure, support selection, and particle size. The role of the perimeter interfaces of Au particles as the sites for reactions is discussed as well as the change in chemical reactivity of Au clusters composed of fewer than 300 atoms. PMID- 12731076 TI - Development of multimedia learning modules for teaching human anatomy: application to osteology and functional anatomy. AB - Computer-assisted learning (CAL) is growing quickly within academic programs. Although the anatomical commercial packages that are available for this learning have attractive advantages, they also have drawbacks: they are frequently not in the local language of the students, they do not perfectly answer the needs of the local academic program, and their cost is frequently more than students can afford. This study describes a relatively inexpensive method to create CAL tutorials, whose content can be fully customized to local academic needs in terms of both program and language. The study describes its use in creating multimedia learning modules (MLMs) about Osteology and joint kinematics. The pedagogical content in these modules was collected from objective experiments to give students the opportunity to access new scientific knowledge during their education. It can be replaced, as desired, by almost any content due to the flexibility of the production method. Each MLM consists of two complementary subelements: a multimedia theoretical lecture and a three-dimensional interactive laboratory. Such MLMs are in use at both the University of Brussels (ULB) and the National University of Rwanda (NUR). The development of this work was part of the VAKHUM project, and the pedagogical validation is currently being performed as part of the MULTIMOD project. PMID- 12731077 TI - Reconstructing cetacean brain evolution using computed tomography. AB - Until recently, there have been relatively few studies of brain mass and morphology in fossil cetaceans (dolphins, whales, and porpoises) because of difficulty accessing the matrix that fills the endocranial cavity of fossil cetacean skulls. As a result, our knowledge about cetacean brain evolution has been quite limited. By applying the noninvasive technique of computed tomography (CT) to visualize, measure, and reconstruct the endocranial morphology of fossil cetacean skulls, we can gain vastly more information at an unprecedented rate about cetacean brain evolution. Here, we discuss our method and demonstrate it with several examples from our fossil cetacean database. This approach will provide new insights into the little-known evolutionary history of cetacean brain evolution. PMID- 12731080 TI - New direction in organopalladium chemistry: structure and reactivity of unsaturated hydrocarbon ligands bound to multipalladium units. AB - Examination of the manner of interaction between Pd(0) and allylpalladium(II) complexes, both being involved as key intermediates in Pd-catalyzed allylic coupling, led us to discover a new role for such combinations in affecting the stereochemistry of the transformations. A similar investigation of the system involving Pd(0) and allenyl/propargyl complexes of Pd(II) led to the discovery of dinuclear Pd(I)bond;Pd(I) complexes containing bridging allenyl/propargyl ligands, which exhibited novel structural and reactivity aspects of great synthetic significance. A systematic comparison was made between the structure, stability, and reactivity of allyl and allenyl/propargyl ligands in dinuclear complexes and those in mononuclear counterparts. On the basis of MO calculations, coordination behavior specific to the ligands of the dinuclear complex is attributed to the occurrence of the back-donating interaction from filled Pdbond;Pd bonding orbitals to vacant ligand pi* orbitals. Similar bonding features are the origin of the ready synthesis of novel one-dimensional sandwich complexes composed of conjugated polyene ligands and linear polypalladium chains. A substitutionally labile dipalladium complex reacts with an equimolar amount of trienes or alkynes to give formal [4pi + 2sigma] or [2pi + 2sigma] adducts, respectively, which undergo further unique transformations with additional unsaturated substrates. PMID- 12731079 TI - Functionalizing inorganic solids: towards organic-inorganic nanostructured materials for intelligent and bioinspired systems. AB - The design, preparation, and properties of organic-inorganic hybrid compounds are described and discussed with respect to their potential uses as intelligent and bioinspired materials. Several synthesis strategies based on intercalation in 2D solids, the grafting of organic groups onto silica and silicates, and the self assembly of organo-silica materials are presented, focusing on the soft procedures that are used to modify the functionality of the inorganic substrates. The combination of both organic and inorganic moieties at the nanometer level forms the basis for preparing multifunctional solids that are provided with specific functions in response to different types of stimuli. In some cases these resemble materials that are found in biological systems. Examples include organic inorganic membranes that are based on intercalated macrocyclic compounds and bi layer vesicles that consist of alkyl long-chains arranged either in the confined region of layered silicates or as self-organized organo-silica micelles. The role of certain hybrid materials such as membranes provides a different approach to the development of artificial liposomes and other mimetic systems that have an organic-inorganic composition and nanostructural organization. Their potential uses for DDS or DNA-dense phases are also discussed and novel alternatives to bioinspired systems development are proposed. PMID- 12731081 TI - Determination of molecular stereochemistry using vibrational circular dichroism spectroscopy: absolute configuration and solution conformation of 5-formyl-cis, cis-1,3,5-trimethyl-3-hydroxymethylcyclohexane-1-carboxylic acid lactone. AB - The determination of the absolute configuration of chiral molecules is an important aspect of molecular stereochemistry. Vibrational circular dichroism (VCD) is the extension of electronic CD into the infrared region where fundamental vibrational transitions occur. VCD has a number of advantages over all previous methods of absolute configuration assignment. The absolute configuration and predominant solution-state conformation in CDCl(3) of the chiral lactone, 5-formyl-cis,cis-1,3,5-trimethyl-3-hydroxymethylcyclohexane-1 carboxylic acid lactone, 1, obtained by the comparison of measured and calculated VCD spectra, are reported. It is found that (-)-1 corresponds to the absolute configuration (1S,3S,5R)-1. PMID- 12731082 TI - Auto-organization in sol-gel type polycondensation: a door to the nanosciences. AB - Silica-based hybrid organic-inorganic materials prepared by sol-gel chemistry exhibit chemical and physical properties revealing their anisotropic organization. Besides the opportunities that these phenomena open for the preparation of new materials, they also provide arguments for the chemist looking for better comprehension and control of the solid's organization. PMID- 12731084 TI - Induction of T cell tolerance to a protein expressed in the cytoplasm through retroviral-mediated gene transfer. AB - BACKGROUND: Host immune responses to foreign gene products have been shown to lead to the elimination of genetically modified cells, and are a major barrier to successful therapeutic gene therapy. We have shown that immunological tolerance to retrovirally transduced cell surface proteins can be induced by expressing the gene encoding these products in bone marrow derived cells. Here, we investigate if expression of foreign gene products in bone marrow derived cells can be used to induce tolerance to cytoplasmic proteins. METHODS: Balb/c mice were reconstituted with syngeneic bone marrow cells transduced with retrovirus carrying the gene encoding enhanced green fluorescent protein (eGFP), or mock transduced bone marrow cells. After reconstitution, mice were immunized with cells expressing eGFP, and T cells were tested for the ability to kill eGFP expressing targets in in vitro cytotoxic T lymphocyte (CTL) assays. RESULTS: T cells from Balb/c mice reconstituted with mock-transduced bone marrow were able to kill target cells expressing eGFP. In contrast, T cells from mice reconstituted with eGFP-transduced bone marrow were unable to kill targets expressing eGFP. In addition, we observed that T cell responses to eGFP in C57BL/6 mice were minimal even under highly immunogenic conditions. CONCLUSIONS: These data suggest that expression of foreign gene products in bone marrow derived cells is capable of inducing T cell tolerance to proteins expressed exclusively in the cytoplasm. PMID- 12731085 TI - Drug-selected co-expression of P-glycoprotein and gp91 in vivo from an MDR1 bicistronic retrovirus vector Ha-MDR-IRES-gp91. AB - BACKGROUND: Retroviral transduction of human hematopoietic stem cells is an attractive strategy in gene therapy; however, transduction efficiency and duration of transgene expression may not be satisfactory in current protocols. Co expression of a human multidrug resistance gene (MDR1) with a therapeutic gene affords selectable growth advantage to genetically modified cells. METHODS: A bicistronic retrovirus vector, Ha-MDR-IRES-gp91, was constructed for the co expression of MDR1 and gp91, a gene responsible for X-linked chronic granulomatous disease (X-CGD). Drug-selected co-expression of P-glycoprotein and gp91 was evaluated in transduced cells. RESULTS: Epstein-Barr virus-transformed B cells from X-CGD patients transduced with Ha-MDR-IRES-gp91 co-expressed human P glycoprotein and gp91, and acquired superoxide-generating activity. Human CD34 positive cells from an X-CGD patient were transduced with Ha-MDR-IRES-gp91 and subsequently treated with 2 ng/ml vincristine. After 13 days, 20% of Ha-MDR-IRES gp91-transduced cells were P-glycoprotein- and gp91-positive by FACS analysis. The superoxide-generating activity of the transduced population was 27% of that of normal cells. Mice transplanted with Ha-MDR-IRES-gp91-transduced bone marrow cells showed co-expression of P-glycoprotein and gp91 in peripheral blood mononuclear cells. By administering paclitaxel, the proportions of P-glycoprotein and gp91-positive cells were increased in all the four mice examined. When mice transplanted with Ha-MDR-IRES-gp91-transduced cells were repeatedly administered paclitaxel, the ratios of P-glycoprotein- and gp91-positive cells were maintained for over 1 year. CONCLUSIONS: These results suggest that MDR1-bicistronic vectors may be useful to select the transduced hematopoietic cells in vivo. This may lead to the sustained expression of transgenes in the blood cells of patients treated with stem cell gene therapy. PMID- 12731086 TI - Local tumour irradiation enhances the anti-tumour effect of a double-suicide gene therapy system in a murine glioma model. AB - BACKGROUND: Gliomas are invasive malignant tumours with poor prognosis. Combination of gene directed enzyme pro-drug therapy with existing treatment modalities might open new therapeutic potentials. METHODS: Murine glioma 261 (Gl261) cells were transduced with an adenoviral vector (Adex-CAUPTK) encoding both uracil phosphoribosyltransferase and thymidine kinase genes which sensitise cells to 5-fluorouracil (5-FU) and ganciclovir (GC), respectively. Subcutaneous or intracranial tumours were established in C57Bl mice by transplanting drug sensitising gene containing Gl261 cells. In vitro growing cells or established tumours were treated with 5-FU, GC and ionising radiation either alone or in combinations. Finally, subcutaneous tumours were established with non-transduced cells, directly injected with Adex-CAUPTK, and mice were treated with combinations of 5-FU, GC and tumour irradiation. RESULTS: In vitro treatment of transduced Gl261 cells with both 5-FU and GC showed enhanced cytotoxic effect compared with single agents. Combination of drug treatments with irradiation greatly increased cytotoxicity. In subcutaneous and intracranial tumours double agent treatment was more effective than a single drug. Combination with local irradiation highly improved the anti-tumour effect (90-100% survival) even when only part of the tumour cells carried drug-sensitising genes (40-50% survival at 10% rate). Treatment of established tumours with direct intra-tumour Adex-CAUPTK inoculations and intraperitoneal 5-FU, GC injections slowed down tumour progression that was further enhanced by local irradiation. CONCLUSION: The combination of double-suicide gene therapy with local irradiation is a promising tool to eradicate small, residual tumours. PMID- 12731087 TI - Concurrent delivery of GM-CSF and endostatin genes by a single adenoviral vector provides a synergistic effect on the treatment of orthotopic liver tumors. AB - BACKGROUND: The immune resistance of large tumors represents a major problem for cancer immunotherapy, whereas the need for repeated injections of high doses of recombinant anti-angiogenic proteins represents a similar problem for anti angiogenic therapy. To test whether antitumor activity could be increased by combining the above two mechanisms, this study examined the therapeutic effect of combination gene therapy using a murine granulocyte-macrophage colony-stimulating factor (mGM-CSF) gene and a human endostatin (hED) gene on a rat orthotopic liver tumor model. METHODS: An adenoviral vector was constructed that simultaneously carried two transcriptional cassettes, for the expression of mGM-CSF and hED, respectively, or that carried a single cassette of either gene. The adenoviruses were intratumorally administered to 3-day-old or 7-day-old tumors. Moreover, the antitumor effects of the combination therapy and monotherapy were assessed and compared. RESULTS: The double-gene-containing adenoviral vector expressed transgenes as efficiently as the single-gene-containing vector. Moreover, the adenovirally expressed endostatin was biologically active, as demonstrated in vitro and in vivo. Results from animal experiments demonstrated a synergistic antitumor effect induced by the combined mGM-CSF and hED therapy. The combination of hED with mGM-CSF enhanced tumor-specific CTL activity, but did not interfere with the infiltration of cellular effectors in the tumor regions. The blood vessel density of the liver tumors markedly reduced as a result of hED expression in both monotherapy and combination therapy. Furthermore, combination therapy significantly increased the number of apoptotic cells in the tumor regions. CONCLUSIONS: The experimental results suggest that the combined gene therapy against tumor cells and the tumor vascular system using antitumor immune mechanisms and anti-angiogenic mechanisms holds promise as a strategy for treating cancers. PMID- 12731088 TI - Multiple muscles in the AMD quail can be "cross-corrected" of pathologic glycogen accumulation after intravenous injection of an [E1-, polymerase-] adenovirus vector encoding human acid-alpha-glucosidase. AB - BACKGROUND: Previously, in murine models of acid maltase deficiency (AMD), we demonstrated that intravenous administration of an improved adenovirus (Ad) vector encoding human acid alpha glucosidase (hGAA) resulted in liver transduction, followed by high-level hepatocyte-mediated secretion of hGAA into the plasma space. The hGAA secreted by the liver was taken up and targeted to muscle cell lysosomes. The levels of hGAA achieved by this approach resulted in clearance of lysosomal glycogen accumulations; in some muscle tissues the effect was prolonged (>6 months). We next wished to demonstrate whether this approach could be generalized across divergent species. To accomplish this goal, we determined whether a similar approach would also result in efficacy, but in a quail model of AMD. METHODS: An [E1-, E2b-]Ad vector encoding hGAA was intravenously injected into AMD quails. At several time points thereafter, plasma, liver, and multiple muscle tissues were assayed for evidence of hGAA gene expression, liver-mediated hGAA secretion, uptake of hGAA by skeletal muscles, and evidence of glycogen correction in AMD skeletal muscles. These results were compared with those obtained from mock-injected AMD or wild-type quails. RESULTS: Intravenous [E1-, E2b-]Ad/hGAA vector injection resulted in high-level liver transduction and hepatic secretion of precursor forms of hGAA. The hepatically secreted hGAA was found to not only be efficiently taken up by cardiac and skeletal muscles, but was also proteolytically cleaved and processed equivalently to the quail-GAA protein detected in wild-type quails. The observations suggest that the signals regulating muscle cell uptake (but not proteolytic cleavage) of lysosomal enzymes are conserved and recognized across divergent species of vertebrates. Importantly, once localized to skeletal muscle lysosomes, the hGAA was able to effectively clear the glycogen accumulations present in AMD quail muscles. CONCLUSIONS: Adenovirus-mediated transduction of the hGAA gene, followed by hepatic secretion, uptake, and cross-correction of the pathologic glycogen accumulation noted in multiple muscles of both the AMD mouse and AMD quail, adds support to the notion that gene transfer strategies (Ad-mediated or other agents) targeting liver tissues with the hGAA gene are likely to be highly efficacious in humans affected by AMD. PMID- 12731089 TI - Preclinical evaluation of DISC-GMCSF for the treatment of breast carcinoma. AB - BACKGROUND: DISC-hGMCSF is a gH-deleted HSV-2 based vector expressing human GM CSF that has entered clinical trials for the therapy of metastatic melanoma. To determine whether this product also has potential to treat breast carcinoma, a series of in vitro and in vivo studies were made. METHODS: Breast carcinoma cell lines and primary cultures of breast carcinoma cells were infected with DISC-GFP or DISC-human-GMCSF (DISC-hGMCSF) and the number of GFP-positive cells and GM-CSF yields were determined. In vivo efficacy of DISC-murine-GMCSF (DISC-mGMCSF) in combination with systemic chemotherapy was assessed in the murine 4T1 breast carcinoma model by direct injection into subcutaneous tumours. RESULTS: DISC hGMCSF was able to infect all breast carcinoma cell lines and the majority of primary breast carcinoma cultures with high efficiency, although culture-to culture variability in infectability was noted in the latter. In the MCF-7 breast carcinoma cell line, expression of hGMCSF was found to peak over the first 24 h post-infection and drop to background levels by 7 to 14 days. In the 4T1 murine breast tumour model, injection of subcutaneous tumours led to a delay in tumour growth and, in rare cases, complete regression of visible tumour. DISC-mGMCSF and DISC-LacZ showed similar levels of efficacy. When mice were given simultaneous 5FU chemotherapy the effectiveness of DISC-mGMCSF treatment was undiminished, and up to three out of ten mice showed complete absence of visible tumour. CONCLUSIONS: DISC-hGMCSF is able to infect human breast carcinoma cells at high efficiency and express GM-CSF. DISC-mGMCSF demonstrated efficacy in the murine 4T1 model, even during concomitant chemotherapy. Taken together these results indicate that DISC-hGMCSF may have potential for the treatment of breast carcinoma. PMID- 12731090 TI - Incorporation of calcium phosphate enhances recombinant adeno-associated virus mediated gene therapy in diabetic mice. AB - BACKGROUND: Increased efficiency of transgene expression is desired for virus mediated gene delivery. In the present study, we examined the effect of calcium phosphate (CaPi) on recombinant adeno-associated virus (rAAV)-mediated insulin therapy in diabetic animals. METHODS: The rAAV vector, rAAV.PEPCK.Ins.EGFP, containing the human insulin gene under control of the phosphoenolpyruvate carboxykinase (PEPCK) promoter and the enhanced green fluorescence protein (EGFP) gene driven by the cytomegalovirus (CMV) IE promoter, was employed in this study. C57BL/6J mice were made diabetic with streptozotocin (STZ), followed by injection into the livers with either rAAV alone, or noncovalent complexes with calcium phosphate. Body weight and blood glucose levels of the animals were routinely monitored after 6 h fasting. Secretion of human insulin in the rAAV-transduced animals was determined by radioimmunoassay (RIA). Expression of human insulin in the livers of the animals was detected by immunohistochemical staining. RESULTS: Compared with the STZ-treated control mice, administration of rAAV containing the human insulin gene significantly decreased blood glucose levels and maintained body weight of the diabetic animals. Complexation of rAAV with calcium phosphate enhanced the hypoglycemic effect of rAAV-mediated gene transfer. Results obtained from both RIA and immunohistochemical staining demonstrated that incorporation of calcium phosphate enhanced rAAV-mediated gene transfer in vivo, leading to higher expression and secretion of human insulin. CONCLUSIONS: Administration of rAAV harboring the human insulin gene into livers of the STZ-diabetic mice improved blood glucose levels, maintained body weight of the diabetic animals, and resulted in human insulin secretion. Complexation of rAAV with calcium phosphate significantly potentiated the efficiency of rAAV-mediated diabetic gene therapy. PMID- 12731091 TI - Gene therapy of streptozotocin-induced diabetes by intramuscular delivery of modified preproinsulin genes. AB - BACKGROUND: Despite improvements in insulin preparation and delivery, physiological normoglycemia is not easily achieved in diabetics. Therefore, there has been considerable interest in developing gene therapy approaches to supply insulin. We studied a nonviral muscle-based method of gene therapy and demonstrated that it could prevent hyperglycemia in murine streptozotocin (STZ) induced diabetes. METHODS: A plasmid encoding mouse furin-cleavable preproinsulin II cDNA (FI), or its B10-analogue (B10FI), and a plasmid encoding furin were coinjected into muscle of CD-1 mice, who were treated a day later with STZ to induce diabetes. Electroporation was applied to increase gene transfer. Blood glucose was measured in fed and fasting mice, and fasting plasma insulin was measured by radioimmunoassay. The form of insulin produced and the presence of C peptide were analyzed by gel filtration chromatography. RESULTS: A B10FI plasmid codelivered with a furin plasmid reduced fed and fasting blood glucose levels in STZ-treated diabetic mice. The (pro)insulin levels in plasma were increased by up to 70-fold versus blank plasmid-treated diabetic mice. The administration of FI with furin was less effective. (Pro)insulin levels were greatly increased by using two plasmids carrying different promoter elements (CMV and SV40). Insulin was identified in muscle cells by immunohistochemistry. In plasma, 40-70% of the (pro)insulin was processed to the mature form and free C-peptide was identified. Insulin gene-treated mice had improved growth rates and appeared healthier. A single injection of B10FI with SV40Furin DNA increased plasma (pro)insulin for at least 8 weeks and reduced fed blood glucose levels for 5 weeks and fasting levels for 8 weeks. CONCLUSIONS: This is the first report that electroporation-enhanced intramuscular gene therapy with B10FI can prevent hyperglycemia in murine STZ induced diabetes. Gene therapy using various routes and methods of furin cleavable insulin gene delivery has been previously explored but, in muscle, results comparable to ours have not been reported. PMID- 12731092 TI - A DNA vaccine containing inverted terminal repeats from adeno-associated virus increases immunity to HIV. AB - BACKGROUND: DNA vaccines have been used to induce both humoral and cellular immune responses against infectious microorganisms. This study explores whether DNA vaccine immunogenicity can be improved by introducing inverted terminal repeats (ITRs) from adeno-associated virus (AAV) into the regulatory region of the DNA plasmid. METHODS: CMV promoter-driven HIV Env expressing plasmid (pCMV HIV) and the pCMV-HIV plasmid introduced ITRs (pITR/CMV-HIV) were transfected in HEK293 cells with LipofectAmine. The HIV Env expression was quantified with Western blot. Fifty micro g of pCMV-HIV or pITR/CMV-HIV plasmid with RIBI adjuvant were immunized to BALB/c mice on days 0, 14 and 28 by intramuscular route, and HIV-specific serum IgG titer was detected 2, 6, 10, 14 and 18 weeks after the first immunization. HIV-specific tetramer assay and HIV-specific IFN gamma ELIspot assay were performed 1 week after the last immunization. The immune mice were intravenously challenged with a vaccinia virus expressing the HIV env gene 1 week after the last immunization. RESULTS: Significantly higher level of HIV Env expression was achieved by pITR/CMV-HIV plasmid. BALB/c mice immunized with pITR/CMV-HIV plasmid generated significantly higher HIV-specific antibody, higher cellular immune responses and lower viral loading than animals immunized with pCMV-HIV plasmid. CONCLUSIONS: AAV ITRs enhance CMV-dependent up-regulation of transgene expression and immunogenicity of DNA vaccine. PMID- 12731094 TI - Comparison of functional brain PET images and intraoperative brain-mapping data using image-guided surgery. AB - OBJECTIVE: Knowledge about the spatial localization of eloquent brain areas is essential for resecting lesions in the vicinity of these areas. The classical approach is to perform surgery on the awake patient under local anesthesia using brain-mapping techniques. As an alternative, the location of eloquent areas can be visualized by preoperative functional brain-imaging techniques, for example, positron emission tomography (PET), functional magnetic resonance imaging (fMRI), or magnetoencephalography (MEG). Using functional activation PET, both methods were combined by integration into a frameless navigation system (BrainLAB) and used to map speech-eloquent areas. PATIENTS AND METHODS: Speech-eloquent areas were localized preoperatively in seven patients with a left-sided glioma using 2 [(18)F]-2-desoxy-D-glucose PET. Patients were scanned under silence conditions (i.e., with the patient remaining silent in a sound-proof cabin), and speech was activated using a verb-generation paradigm. The PET data were transferred to the neuronavigation workstation and matched with a preoperative 3D-MRI using an automatic image-fusion algorithm. Intraoperative speech localization was performed using brain-mapping techniques under local anesthesia with bipolar cortical stimulation. The stimulator position was mapped into the MRI/PET data set by neuronavigational tracking of the instrument. RESULTS: Functional PET images were integrated into the MRI-based neuronavigational system and could be transferred exactly to the operative field. By the additional integration of cortical stimulation, intraoperative electrophysiological findings can be directly compared with preoperative functional images. Seven patients with left sided glioma were operated on using this protocol, confirming the technical feasibility. In three of seven patients, preoperative PET findings were not supported by intraoperative mapping. CONCLUSIONS: This matching and mapping technique is suitable for monitoring eloquent speech areas during surgical resection of extensive left-sided low-grade gliomas, allowing a direct comparison between intraoperative electrophysiological brain mapping and preoperative functional brain-imaging findings. The sensitivity and specificity of functional imaging techniques can now be evaluated by reconciling the data with the intraoperative stimulation results. PMID- 12731095 TI - Use of three-dimensional computer graphic animation to illustrate cleft lip and palate surgery. AB - OBJECTIVE: Three-dimensional (3D) computer animation is not commonly used to illustrate surgical techniques. This article describes the surgery-specific processes that were required to produce animations to teach cleft lip and palate surgery. MATERIALS AND METHODS: Three-dimensional models were created using CT scans of two Chinese children with unrepaired clefts (one unilateral and one bilateral). We programmed several custom software tools, including an incision tool, a forceps tool, and a fat tool. RESULTS: Three-dimensional animation was found to be particularly useful for illustrating surgical concepts. Positioning the virtual "camera" made it possible to view the anatomy from angles that are impossible to obtain with a real camera. Transparency allows the underlying anatomy to be seen during surgical repair while maintaining a view of the overlaying tissue relationships. Finally, the representation of motion allows modeling of anatomical mechanics that cannot be done with static illustrations. The animations presented in this article can be viewed on-line at http://www.smiletrain.org/programs/virtual_surgery2.htm. CONCLUSIONS: Sophisticated surgical procedures are clarified with the use of 3D animation software and customized software tools. The next step in the development of this technology is the creation of interactive simulators that recreate the experience of surgery in a safe, digital environment. PMID- 12731096 TI - Computer-based periaxial rotation measurement for aligning fractured femur fragments from CT: a feasibility study. AB - A new computer-based method for measuring periaxial rotation of healthy and fractured femurs from preoperative CT during closed femoral fracture reduction surgery is described. The method provides a comparative quantitative measure to align the distal and proximal femur fragments based on periaxial rotation. The periaxial rotation is defined in terms of patient-specific bone features. An algorithm for automatically extracting these features from the CT based on this definition has been developed. The algorithm extracts the condyle landmarks and neck axis of the healthy bone, determines its periaxial rotation, and extrapolates this data, assuming mirror symmetry between the healthy and fractured bones, to measure periaxial rotation between the fractured fragments. Unlike existing techniques, the method requires minimal user intervention. In a feasibility study, the method was applied to five dry femurs and one patient data set, and simulated a reduction based on the periaxial measurements with satisfactory results. The experiments showed the measured angle on the fractured femur to be within 1-4.5 degrees of that of the healthy bone. PMID- 12731097 TI - Three-dimensional image registration of phantom vertebrae for image-guided surgery: a preliminary study. AB - OBJECTIVE: Applications of three-dimensional ultrasound (3D US) are emerging throughout the field of medicine. In this study, tracked, free-hand 3D phantom US images were mapped to computed tomograms (CT) as a development for image-guided surgery (IGS) of the spine. In the operating room, the registration of tracked 3D US images to other imaging modalities, such as CT, could allow the surgeon to identify more precisely the surgical target area prior to the incision. An independent quantitative measure of registration accuracy using a fiducial marker system was provided. METHODS: Three-dimensional free-hand US images of a phantom spine were created by tracking the transducer with an optical sensing system. Two sets of images were acquired from three lumbar vertebrae using 4.5- and 7.5-MHz transducers. These images were then segmented for the extraction of the posterior vertebral surface. Next, a surface-based registration of US to the corresponding segmented CT images was performed. Registration errors were computed as the distance between a set of target points transformed using the experimental transformation and the same set of target points transformed using fiducial markers as a gold standard. RESULTS: Results indicated that alignment of these image sets is feasible using only part of the vertebral surface. In particular, the regions of the spinous process and laminae were used for registration. Target registration errors (TREs) were found to be lowest using the highest resolution CT images. Using the CT scans with 2-mm slice thickness, the TRE was calculated to be 1.33 +/- 0.30 mm for the 7.5-MHz US data set and 2.81 +/- 0.10 mm for the 4.5-MHz US data set. Moreover, residual errors in these surface alignments were 0.69 +/- 0.18 mm and 0.61 +/- 0.20 mm for the 4.5- and 7.5-MHz sets, respectively. CONCLUSION: A rigid, surface-based registration of CT images to phantom spinal US images, acquired with a free-hand, tracked transducer, is achievable with a limited, easily obtainable portion of the vertebral surface. PMID- 12731099 TI - Real-time measurement of blood vessel occlusion during microsurgery. AB - Measurement and feedback of vascular properties during microsurgery is generally not available. We carried out real-time in vivo measurement and analysis of microsurgical occlusion of 1-2-mm diameter arteries and veins in rodents. A pair of forceps mounted with strain gauges was designed for applying and directly measuring the force on tissue. Forces between 0 and 450 mN were applied, with the device having a resolution of 0.5 mN. We performed in vivo experiments on the rat femoral (n = 5) and abdominal (n = 8) blood vessels to measure the elastic restoration force of the tissue in response to radial compression at different levels of force. On average, the minimum occlusion force was 57 mN for the rat artery. During steady application of force, the perturbations in the blood vessel due to heartbeat are visible in the force data. These force oscillations ranged between 1 and 3 mN around the mean steady-state force applied. It was determined that the magnitude of the Fourier spectral peak corresponding to heartbeat frequency can be used as a measure of the patency of the blood vessel, and can provide feedback to microsurgeons to avoid damage to the vessel by application of excess force. PMID- 12731098 TI - Spine needle biopsy simulator using visual and force feedback. AB - OBJECTIVE: Biopsy with an inserted needle is an important procedure for lesion detection in the spine, but is difficult to perform due to the presence of many critical organs near the spine. This article presents a spine needle biopsy simulator, based on visual and force feedback, which can be used to plan the optimal path of a needle and to practice the procedure without risk. MATERIALS AND METHODS: The simulator is composed of a 3D human model, a visual-feedback component, a force-feedback component, and an evaluation module. The human model is based on 3D CT data. The visual-feedback component provides an oblique section, multiplanar reformatting images, and a volume-rendered image. Of these, the oblique section display is very useful for planning a 3D path for the needle. During simulation, the force-feedback component generates and provides realistic forces acting on the biopsy needle in real time by synchronizing them to visual feedback. After each simulation, the evaluation module provides a performance analysis for the trainee. RESULTS: For an XCT abdomen volume data set of 256 x 256 x 256, the update rate of image rendering due to needle movement is over 25 Hz, with a force-feedback rate of 1 kHz. This performance proved to be good enough for the trainee to learn the relationship between visual and force feedback. CONCLUSIONS: The simulator is useful for the planning of and training in complicated 3D spine needle biopsy procedures. It may be used as an educational tool for beginners, a practice tool to increase expertise, or a test bed for new procedures. PMID- 12731100 TI - Robust Bayesian estimation of the hemodynamic response function in event-related BOLD fMRI using basic physiological information. AB - In BOLD fMRI data analysis, robust and accurate estimation of the Hemodynamic Response Function (HRF) is still under investigation. Parametric methods assume the shape of the HRF to be known and constant throughout the brain, whereas non parametric methods mostly rely on artificially increasing the signal-to-noise ratio. We extend and develop a previously proposed method that makes use of basic yet relevant temporal information about the underlying physiological process of the brain BOLD response in order to infer the HRF in a Bayesian framework. A general hypothesis test is also proposed, allowing to take advantage of the knowledge gained regarding the HRF to perform activation detection. The performances of the method are then evaluated by simulation. Great improvement is shown compared to the Maximum-Likelihood estimate in terms of estimation error, variance, and bias. Robustness of the estimators with regard to the actual noise structure or level, as well as the stimulus sequence, is also proven. Lastly, fMRI data with an event-related paradigm are analyzed. As suspected, the regions selected from highly discriminating activation maps resulting from the method exhibit a certain inter-regional homogeneity in term of HRF shape, as well as noticeable inter-regional differences. PMID- 12731101 TI - Cerebral information transfer during word processing: where and when does it occur and how fast is it? AB - Different regions of the brain have to interact to perform language processing. Such neural integration processes can be studied by measuring synchronization of oscillations. Coherence is the best known algorithm to study synchronization between two sites of the cortex with regard to selected oscillation frequencies. Phase coherence quantifies phase synchronization, in particular. In addition, cross phase (or simply phase) gives information about the direction and speed of the spread of oscillations. We use a new method to study short-time phases between different sites of the cortex in order to explore transient neural networks during word processing. Particularly, processing of abstract and concrete nouns was investigated by dynamic phase analysis of the alpha1 frequency band (8-10 Hz). Abstract and concrete nouns turned out to induce different dynamic networks of information transfer. Whereas processing of concrete nouns excites a widespread network for about 800 msec, the interhemispheric interactions during abstract noun processing are restricted to 300-500 msec after word presentation. Further, the direction of cerebral information transfer differs for the two word categories. Additionally, it could be shown that the propagation speed of information is slower for concrete nouns than for abstract nouns. PMID- 12731102 TI - Demand on verbal working memory delays haemodynamic response in the inferior prefrontal cortex. AB - Event-related functional magnetic resonance imaging was used to test the involvement of the inferior prefrontal cortex in verbal working memory. Pairs of French nouns were presented to ten native French speakers who had to make semantic or grammatical gender decisions. Verbal working memory involvement was manipulated by making the categorization of the second noun optional. Decisions could be made after processing the first noun only (RELEASE condition) or after processing the two nouns (HOLD condition). Reaction times suggested faster processing for gender than for semantic category in RELEASE. Despite the absence of anatomical difference across tasks and conditions in the wide activated network, the haemodynamic response peak latencies of the inferior prefrontal cortex were significantly delayed in HOLD versus RELEASE while no such peak delay was observed in the superior temporal gyrus. Interestingly, this pattern did not interact with language tasks. This study shows that cognitive manipulation can influence haemodynamic time-course and suggests that the main cognitive process determining inferior prefrontal activation is verbal working memory rather than specific linguistic processes such as grammatical or semantic analysis. PMID- 12731103 TI - Brain activations during motor imagery of locomotor-related tasks: a PET study. AB - Positron emission tomography (PET) was used to study the involvement of supraspinal structures in human locomotion. Six right-handed adults were scanned in four conditions while imagining locomotor-related tasks in the first person perspective: Standing (S), Initiating gait (IG), Walking (W) and Walking with obstacles (WO). When these conditions were compared to a rest (control) condition to identify the neural structures involved in the imagination of locomotor related tasks, the results revealed a common pattern of activations, which included the dorsal premotor cortex and precuneus bilaterally, the left dorsolateral prefrontal cortex, the left inferior parietal lobule, and the right posterior cingulate cortex. Additional areas involving the pre-supplementary motor area (pre-SMA), the precentral gyrus, were activated during conditions that required the imagery of locomotor movements. Further subtractions between the different locomotor conditions were then carried out to determine the cerebral regions associated with the simulation of increasingly complex locomotor functions. These analyses revealed increases in rCBF activity in the left cuneus and left caudate when the W condition was compared to the IG condition, suggesting that the basal ganglia plays a role in locomotor movements that are automatic in nature. Finally, subtraction of the W from the WO condition yielded increases in activity in the precuneus bilaterally, the left SMA, the right parietal inferior cortex and the left parahippocampal gyrus. Altogether, the present findings suggest that higher brain centers become progressively engaged when demands of locomotor tasks require increasing cognitive and sensory information processing. PMID- 12731105 TI - Cattle transport regulations in the European Union. AB - The essential items of the relevant EU legislation are described and their effects are discussed. A brief survey of the four most important legal regulations in the EU relevant for cattle transport is given from the point of view of administration. These regulations are: The Directive 91/628/EEC 1991 (protection of animals during transport), Council Regulation (EC) No 1255/97 (criteria for staging points and route plan), Council Regulation (EC) No 411/98 (additional standards for road vehicles for long distance transport), Commission Regulation (EC) No 615/98 (export refund regulations). Some critical points of the objectives are pointed out. PMID- 12731104 TI - Transport stress in cattle and sheep with details of physiological, ethological and other indicators. AB - The welfare of animals during transport should be assessed using a range of behavioural, physiological and carcass quality measures. In addition, health is an important part of welfare so the extent of any disease, injury or mortality resulting from, or exacerbated by, transport should be measured. Many of the indicators are measures of stress in that they involve long-term adverse effects on the individual. Key factors affecting the welfare of animals during handling and transport which are discussed are: attitudes to animals and the need for training of staff; methods of payment of staff; laws and retailers' codes; genetics especially selection for high productivity; rearing conditions and experience; the mixing of animals from different social groups; handling procedures; driving methods; stocking density; increased susceptibility to disease and increased spread of disease. PMID- 12731106 TI - Animal transport needs reasonable legislation. AB - The paper describes practical problems of animal trade. It is written from the point of view of an animal breeding organisation and does therefore not discuss in detail physiological indicators or the development of the EU legislation in that field. Based on the practical experiences it is assumed that animals can be transported in a reasonable and responsible way, without significant and long lasting negative effects, even over long distances. The current rules for animal transport in the EU, which have been developed between 1991 and 1998 still show serious shortcomings as they appear to have been laid down prematurely and in the absence of sufficient scientific knowledge. The paper points to some of the shortcomings and provides for proposals to change the current legislation: Unloading and loading at staging points should be dropped, loading densities should be derived from practical experience, driving and resting times for animals and driver must be harmonised, the regulation of staging points needs to be simplified and the control and sanctioning system must be uniform and transparent. PMID- 12731107 TI - Handling of slaughter cattle in pre and post transport situations including loading and unloading on journeys up to 8 hours in Germany. AB - In a field study 63 commercial cattle transports to German slaughter plants including 580 bulls, cows and heifers were investigated to estimate the impact of pre and post transport aspects on animal welfare. Study 1 evaluated these aspects as part of a large model covering all possible influences at transport and slaughter on biochemical stress response and carcass and meat quality. Included were keeping system, behavioural and clinical observations at loading, unloading and lairage, transport time, loading density, mounting prevention, regrouping and lairage time. In study 2 more detailed questions about loading and unloading were answered by investigating the impact of facility design and handling criteria on cattle behaviour and heart rate. Cattle from tied housing show an increased stress response to all transport procedures. Many aspects of facility design and handling contribute to quality of loading. Cattle show calmer behaviour, turn, balk or fall less if they are not mixed and driven in small groups on driveways without impediments but with regular sideblock, not wider than the vehicle ramp. Vehicle ramps should have high and closed side gates, be inclined less than 20 degrees, and the step towards ramp should be less than 15 cm. Unloading is improved if performed level and to the back of the vehicle (behaviour, heart rate, cortisol, CK). Direct reloading should be avoided and mounting should be prevented during transport and lairage (CK, lactate, bruising). But mounting prevention devices may increase bruising if set too low (< 20 cm above withers). Longer lairage time is disadvantageous (CK, energy metabolism). PMID- 12731108 TI - Assessment of air quality in a commercial cattle transport vehicle in Swedish summer and winter conditions. AB - Transport by road can induce significant stress in cattle. Thermal stress is among the main stress producing factors during transport. The provision of ventilation in livestock transport vehicles is usually through openings along the sides of the vehicle. The incoming air will affect air quality inside by regulating temperature, relative humidity, gas levels and levels of other contaminants. The aim of the present investigation was to map out the air quality in a commercial cattle transport vehicle under various climatic conditions and with varying stocking densities and transport times. Distributions of air temperature, relative humidity and concentrations of ammonia, carbon dioxide, oxygen and methane have been determined during 35 experimental journeys. In average the mean temperature inside the compartment was about 3 degrees C and 6 degrees C higher than outside temperature in summer (+7.8(-)+24.0 degrees C) and winter (-24.3(-)+12.7 degrees C) conditions respectively. The temperature increment inside, as could be expected from theory, increased with reduced ventilation and increased animal density. Many stops to load new animals lowered the temperature increment and relative humidity in winter time. In summer more stops made the compartment temperature and relative humidity increase. The inside temperature distribution was less than about 3 degrees C during both summer and winter season. Average ammonia level varied between 3 and 6 ppm depending on stocking density and number of stops with a maximum value of 18 ppm. No detectable methane levels could be found inside the compartment at any time. PMID- 12731110 TI - Effect of journey time and environmental condition on bull behaviour and beef quality during road transport in northern Italy. AB - Response to a transport time of 1 and 3 hours on the road carried out under different environmental conditions was evaluated on 60 pure-breed Charolaise bulls. Temperature and relative humidity from loading to slaughter were expressed as THI (Temperature-Humidity Index). Regardless of journey time or environmental conditions, the most common standing orientation was either perpendicular or parallel to the direction of travel. Diagonal orientation increased with THI class. Cortisol and glucose plasma concentrations increased because of transport but they were not influenced by journey time or THI class. The CK plasma concentration, instead, increased according to THI class. The incidence of carcass bruising was not affected by the journey time or by the environmental conditions. Journey time also had a negligible effect on beef quality while, meat lightness increased and water holding capacity slightly decreased depending on THI. PMID- 12731109 TI - Effect of commercial transport in Spain on cattle welfare and meat quality. AB - The purpose of this work was to determine the effect of transport of cattle from the farm to the abattoir on the animal's welfare and post mortem meat quality (instrumental and sensorial). Thirty-nine commercial journeys were followed in Spain from July 2000 to July 2001 at varying distances from a local abattoir in the city of Zaragoza. Blood samples were taken at the farm and at sticking and meat pH24 was measured at 24 hours post-mortem. Samples from the M. longissimus dorsi thoracis were taken from 48 slaughter bulls and analysed in terms of instrumental and sensorial qualities. Cortisol, glucose, CPK, lactate and N:L ratio increased during transport compared to control values. Cortisol was higher after 1-2 h transport compared to journeys that were less than 1 h or more than 2 h long. In general the parameters indicated slightly more stressful conditions under winter conditions than in the summer. Meat tenderness measured by a Warner Bratzler device demonstrated that there was no significant effect of either transport time or season. There was no significant change in pH24 or meat colour at 24 h with transport time. Transport time had a significant effect (p < 0.05) on overall liking and animals transported in summer had slightly more tender meat and higher overall liking. PMID- 12731111 TI - The impact of vertical vibrations on the welfare of calves. AB - The objective of this study was to evaluate the heart rate, body temperature, saliva cortisol levels and behaviour of young calves during transport simulation, and to define comfort conditions for calves related to the frequency and acceleration of vibration. Calves with an average age of 22 days were vibrated in the vertical direction for 2 hours at 2, 4, 8 and 12 Hz, in combination with root mean square (RMS) acceleration magnitudes 1 or 3 m/s2. Welfare and stress were quantified by measuring changes of heart rate, body temperature, cortisol concentration in saliva and behaviour. Treatments with acceleration 3 m/s2 had the largest impact on the animals. Stress response was larger in combination with 2 Hz, especially during the first hour of the experiment. Treatment 12 Hz in combination with acceleration 3 m/s2 had initially no influence on the animals, but towards the end of the treatment the calves started to express a stress response. From these results we can conclude that certain vibrations are stressful for calves and those can impair the welfare during transport. PMID- 12731112 TI - Dynamic performances of cattle transporting vehicle on Scandinavian roads and behavioural response of animals. AB - Driving performance, air quality in the vehicle, and handling during loading and unloading are the main factors, which could affect animal welfare. During transport of animals from farms to abattoirs, animals on the vehicle are subjected to vibrations in all directions. Performances of a typical vehicle for cattle transport have been studied under field commercial conditions. During the experiment, tri-axial vibrations on the vehicle, as well as velocity, slope and position were measured. The animals were video recorded for behavioral analyses. Geographical positions, speed and slope of roads were recorded using, GPS. Measurements were made with and without animals on the vehicle. The roads in the region are narrow almost everywhere, but rather plane and of good quality. Three road types were identified: straight and plain, curvy and graveled roads. Events such as sudden stops and curves were observed and noted. Vibration levels in tri axial directions have been measured and analysed for different speeds and road types both when the vehicle was fully loaded and unloaded. There is a significant difference between the performances of the vehicle when loaded and unloaded. Very high vibrations values have been noted during the empty driving and these values were reduced by up to 9 times when the vehicle was loaded. Less vibrations amplitudes were observed and animals were calmer on ferry transport than on road transport. It has also been noted that animals prefer to stand perpendicular to the direction of vehicle's motion during transport. PMID- 12731113 TI - Special problems of long-distance road transports of cattle. AB - Physiological and behavioural field studies on long distance road transport of 116 heifers, 135 bulls and 64 steers in 10 goose-necked double decked semitrailers from northern Germany to mediteranean ports showed different impacts of handling during pre-transport (i.e. collection of animals, weighing, loading), transport journey itself and post transport handling (i.e. lairage time) on coping strategies of the different categories of cattle. It was found loss of body weight in steers (-6.65%) coming from pasture was higher compared to bulls ( 4.6%) during transport, but they recovered during lairage time in a better way. All categories of cattle showed catabolic energy metabolism during transport, but only in bulls and to a farer extent in heifers this leads to a tendency of a ketotic metabolism during second parts of transport and lairage time. During whole transport time no more than 20% of bulls and steers were laying down, and less than 5% were feeding during driving intervals. In all parts of transport general stress parameters like heart rate (with exception to steers) and cortisol were elevated as a part of adaptation to the transport environment, but indicating high physical and emotional loads on the animals with no resting possibilities. In this context animals have to be prepared carefully to be transported, i.e. in reference to energy and fluid balance, and to be feed in sufficient time intervals (breaks) and lengths to maintain fundamental behavioural and physiological needs of the animals during transport. The lairage facilities are very important for the bulls and in the case of heifers the feeding regime during lairage time must be improved to ensure the possibility for a real resting and recovery of the animals after transport. PMID- 12731114 TI - Meat quality and transport stress of cattle. AB - This study evaluated the effect of transport time up to 14 hours and the effects of vehicle design on animal welfare, stress and meat quality. 18 transports (six short, medium and long) with a total of 486 animals (118 sample animals, heifers and bulls) were carried out on commercial vehicles in summer 2000 and winter 2001. Animal welfare and stress were evaluated by blood serum parameters, heart rate monitoring, behaviour recording and occurrence of carcass bruising. Meat quality was evaluated by post mortem muscle glycogen content, pH value, temperature, drip loss, colour and tenderness measurements. Heifers had lower heart rates than young bulls during loading (95 vs 114 beats per minute, bpm), whereas during transport, both had an average heart rate of 100 bpm, furthermore during unloading, heifers had higher heart rates than bulls (109 vs 100 bpm). Blood sampling during unloading could have marginally increased heart rates during the unloading procedure. Studied cattle had lower heart rates during medium and long distance transports compared with short transports. Monitoring of animal behaviour during transport showed that the former settled down faster than the latter. Single- and two-animal pens in medium and long distance vehicles prevented nervous and stressful movements of cattle, which were more prominent in large pens of short distance lorry. Present results suggest that larger pens of three or four animals could increase cattle stress during transport. Moreover during unloading, cattle loaded in single- or two-animals pens had significantly lower blood cortisol content than those loaded in larger groups of three or four animals (P < 0.01). The amount of severe carcass bruising was highest in animals transported over short times and loaded into groups of four cattle. Severe damages occurred most often on perianal and hipbone area of the carcass surface. Present results showed that muscle glycogen level was highest after long transport. These animals were fed more regularly from the last feeding up to stunning than medium or short distance animals. Animals in single-pens had the highest muscle glycogen level. Transport distance or number of animals in one pen had a minor effect on muscle pH values or temperatures during 24 hours post mortem (pm). Drip loss of the M. longissimus dorsi (LD) was highest after long transport, but animal number in one pen had no effect on drip loss. Colour of the LD muscle was independent on transport conditions. Light colour of three animal groups resulted from high amounts of heifers, which had lighter colour than bulls. All meat samples were quite tender. However, heifers had significantly tender meat than young bulls (P < 0.001). Higher amounts of heifers had the most tender meat after short transports. Mean DFD (dark, firm, dry) meat occurrence was 2.1% in this project, DFD frequency was lowest after short, then after long and highest after medium distance transports. Because of not evenly distributed numbers of bulls (low) and heifers (high) it was difficult to compare short and long distance transport effects. PMID- 12731115 TI - Recommendations for future development in cattle transport in Europe. AB - Every year millions of calves and cattle are transported across, from and to Europe. Most of these animals are going to slaughter houses in the respective countries or in another community state or coming or going abroad (extra-EU). These transports give cause for concern for at least three reasons: First, it can cause severe stress in animals entailing poor welfare. Second, stressful transports may have a negative effect on meat quality. Third, there is the risk of spread of infectious diseases over large distances. Existing legislation does not provide enough protection to transported animals especially over long distances largely because considerable parts of the regulations are not sufficiently based on scientific evidence. In recent years some research is carried out including the EU financed CATRA research project (contract QLK5-CT 1999-0157) concentrating on the welfare and meat quality aspects of cattle transport. This paper summarises important results of this recent research and gives some recommendations for future legislation. The welfare of the animals is limited by their needs not by a fixed maximum transport time, if vehicle and transport conditions are appropriate. Bulls, steers and heifers are reacting differently on transport. Adapt transport schemes to the needs of the animals. Meat quality is only effected in extreme situations. Some animals develop an energy deficit after 6 h of transport. Develop appropriate feeding regimes for long transport. Abolish stressful loading and unloading in staging posts (injuries, infectious diseases). Staging posts are particularly stressful for bulls. Educate handlers and drivers more intensively. Pay drivers inverse to losses. Develop monitor systems for long and short distance transport (e.g. records, GPS). Improve vehicle design (e.g. vibration). PMID- 12731116 TI - 5th IUHPE European Conference on the Effectiveness and Quality of Health Promotion. PMID- 12731117 TI - The politics of reforming health policies. PMID- 12731118 TI - Challenges in health promotion policies. PMID- 12731119 TI - How is success judged in education? PMID- 12731121 TI - IUHPE/CDC: global health promotion and health education initiatives related to chronic disease prevention. PMID- 12731120 TI - Using the Internet as a phone service: an improved technology that can benefit health promotion and education. PMID- 12731122 TI - Global activities of the National Center for Chronic Disease Prevention and Health Promotion (NCCDPHP), Centers for Disease Control and Prevention, USA. PMID- 12731123 TI - [The metabolic syndrome]. AB - Alterations in the intermediary metabolism respectively a "metabolic maladaptation" to the modern lifestyle represents an important risk factor for most chronic diseases, especially also for cardiovascular risk. An increased concentration of free fatty acids plays a central role in the pathophysiological sequence of events. Overweight and obesity represent one important cause for the increase in free fatty acids and are thus potentially preventable causes. It seems that the metabolic syndrome represents the result of our genetic predisposition as a response to the modern lifestyle characterized by physical inactivity and a increase in the duration of the postprandial status. PMID- 12731124 TI - [Medical problems in migrants]. AB - Medical problems in migrants are not only an abstract political problem but also a problem that concerns physicians in their daily practice. Beside somatic disorders that are more common in developing countries we often are confronted to problems due to psychosocial stress. Language difficulties, problems of cultural misunderstanding and a lack of successful concepts impair the physician-patient relationship and hinder a satisfying solution. Missing economic resources influence the availability of efficient support. Despite all difficulties there are strategies that can help practitioners to deal with the subject without waiting for a social revolution. PMID- 12731125 TI - [Doctor-patient interaction in the context of migration]. AB - Doctors are often challenged by their interaction with migrants. The difficulties they describe are essentially the same as the ones they experience with Swiss patients. However, these encounters are frequently encumbered by factors relating to the migrant's situation. This article will delineate the various problem areas of doctor-patient interactions in the context of migration: difficulties understanding one another lack of mutual reference points between doctors and patients and structural frameworks. These problem areas will be discussed and concrete solutions and practical recommendations will be outlined. PMID- 12731126 TI - [Difficult management of a patient with type 2 diabetes mellitus]. PMID- 12731128 TI - The thoroughly modern consultant? Modernizing consultant careers. PMID- 12731127 TI - [Iatrogenic dementia?]. PMID- 12731129 TI - Obtaining and using human tissues for research: ethical and practical dilemmas. PMID- 12731130 TI - Liver transplantation. AB - Liver transplantation has evolved from an experimental procedure to an acceptable therapy for end-stage liver disease. The major challenges now faced are donor organ shortage, long-term complications related to immunosuppressive therapy, and the prevention and treatment of disease recurrence. PMID- 12731131 TI - Bone transplantation. AB - Bony deficiency, particularly loss of bone stock associated with failed joint replacements or tumours, is a challenging problem in orthopaedic surgery. Bone transplantation techniques provide solutions that can be tailored to the clinical problem. However, the risks of bone transplantation are well documented and the biology of allograft incorporation remains unpredictable and poorly understood. PMID- 12731132 TI - The second stage of labour. AB - Obstetric intervention in the second stage of labour is frequently indicated. An appreciation of the physiology of the second stage and an awareness of the range of interventions with their appropriate selection and application will contribute towards ensuring a safe delivery. PMID- 12731134 TI - Inhaled foreign bodies in children: diagnosis and treatment. AB - An inhaled foreign body in children is a rare but potentially fatal condition. This article reviews this clinical condition, in particular highlighting the high index of suspicion needed on behalf of the clinicians involved in the care of children. The literature relating to this condition is reviewed and a summary of the clinical course is presented. PMID- 12731133 TI - Tacrolimus ointment for the management of atopic dermatitis. AB - The management of atopic dermatitis is set for a major shift with the introduction of the first new topical treatment for the condition in 40 years- the non-steroidal topical immunomodulator tacrolimus (Protopic). Backed by strong clinical data, tacrolimus ointment is a valuable addition to topical steroids in the management of moderate-to-severe atopic dermatitis. PMID- 12731135 TI - Evidence-based practice in urogynaecology. AB - Urogynaecology practice is rapidly progressing, with a trend towards minimally invasive techniques. This article attempts to cover the evidence behind the assessment and treatment of urinary and anal incontinence, and vaginal prolapse. PMID- 12731136 TI - The current status of Paget's disease of the bone. AB - Paget's disease is a metabolic bone disease characterized by abnormalities of bone turnover, structure and architecture. The disease is of unknown aetiology, although both genetic and environmental factors have been implicated. Treatment is indicated for patients with active disease and currently bisphosphonates are the first-choice management option. PMID- 12731137 TI - The eye in endocrine disease. PMID- 12731138 TI - A rough guide to research for senior house officers and specialist registrars. AB - Although not mandatory, a period of formalized research leading to a higher degree is still commonplace for doctors wishing to pursue a career in hospital medicine. This may be undertaken before or during specialist registrar training. This article provides practical information for those planning to undertake research. The potential pitfalls of research are discussed and strategies are suggested to ensure that this phase of medical training is both productive and fulfilling. PMID- 12731139 TI - Difficulties in the diagnosis of acute aortic dissection. PMID- 12731140 TI - Angiomyofibrosarcoma: a rare ischiorectal fossa swelling. PMID- 12731141 TI - Pseudoangiomatous hyperplasia of mammary stroma: an unusual entity. PMID- 12731142 TI - Peripartum cardiomyopathy in Nigeria. PMID- 12731143 TI - Clinical electrophysiology. PMID- 12731144 TI - Can simulators be used to assess clinical competence? PMID- 12731145 TI - A brief history of behavioral and cognitive behavioral approaches to sexual offenders: Part 1. Early developments. AB - This is the first of two papers which briefly outline the development of behavioral and cognitive behavioral treatment of sexual offenders from the mid 1800s to 1969. We first consider the historic role of Sigmund Freud and note that a broad scientific interest in deviant sexual behaviour was well established by 1900. In the early to mid-20th century, two psychologies were prominent in the development of behaviorial approaches, those of John B. Watson and Alfred Kinsey. Behavior therapy for a variety of problems emerged in the 1950s and soon found application to deviant sexuality. The development of penile plethysmography helped to focus interest on deviant sexual preference and behavior. While nonbehavioral approaches to sexual offenders paralleled these developments, a combination of behavioral and cognitive behavioral treatments began to emerge in the late 1960s which ultimately developed into the approaches more commonly seen today. PMID- 12731146 TI - A brief history of behavioral and cognitive behavioral approaches to sexual offender treatment: Part 2. The modern era. AB - This paper is the second in our attempt to highlight the major influences, trends, and emerging issues in cognitive behavioral approaches to treating sexual offenders. In this paper the developments from 1970 until the present are covered. PMID- 12731147 TI - Implications for treatment of sexual offenders of the Ward and Hudson model of relapse. AB - A group of 59 child abusers were classified as having either an "avoidant" (n = 15) or an "approach" (n = 44) goal regarding deviant sexual activity with children. Level of distorted beliefs about sexual activity with children (cognitive distortions) and distorted beliefs about their own victims (victim blaming attitudes) in both groups were measured before and after treatment. Results indicate that there was an overall reduction in the level of these distorted beliefs at the posttreatment stage in the approach group. Reductions were not found in the avoidant offenders as men in this group did not have distorted beliefs prior to treatment. Results are discussed in terms of appropriate targeting of treatment. PMID- 12731148 TI - College women who had sexual intercourse when they were underage minors (13-15): age of their male partners, relation to current adjustment, and statutory rape implications. AB - In a survey of 1,439 female college students, 24% reported that they had what they considered consensual sexual intercourse between ages 13 and 15 (2% at age 13, 7% at age 14, and 15% at age 15). Contrary to the impression left by studies of teenage mothers, the majority of their male sexual partners were not substantially older than them but instead were more typically "somewhat older" (2 4 years apart) or similar aged (less than 2 years apart). The percentage of "much older" partners (5 or more years older) was 31% for those who had intercourse at age 13, 17% for those who had intercourse at age 14, and 13% for those who had intercourse at age 15. Women who had intercourse at age 13 endorsed more current symptoms of psychological distress than those who first had intercourse at age 14 or 15. There were no significant differences between the groups in current levels of sexual satisfaction. Partner's age difference was not significantly associated with current levels of either psychological distress or sexual satisfaction. The implications of these results were discussed in light of recent calls in the United States for more strict and rigorous enforcement of statutory rape laws. PMID- 12731149 TI - Predicting psychological distress in sex offender therapists. AB - The provision of psychological services to sexual offenders presents therapists with many challenges, including exposure to vivid descriptions of sexual violence and trauma. Although there is an increasingly robust body of literature investigating the effects of such traumatic exposure on therapists who work with the victims of sexual abuse, there have been few studies of its impact on those who treat the perpetrators. This study provides an exploratory investigation of the experience of psychological distress among therapists who work with sex offenders, as well as the social support mechanisms that may serve to mitigate therapist risk. Fifty-nine sex offender therapists completed a self-report measure of posttraumatic stress symptomatology, and rated their perceptions of family and peer support. therapists also reported the number of hours that they devote to sex offender treatment and the amount of clinical supervision they participate in on a weekly basis. As a group, participants reported experiencing low levels of general and trauma-related distress. Although statistical analyses failed to indicate the existence of a significant positive correlation between number of hours devoted to sex offender treatment and the experience of therapist distress, greater perceptions of peer support were significantly predictive of lower levels of psychological distress and PTSD symptoms. These results are discussed relative to their implications for sex offender therapist self-care and future research. PMID- 12731150 TI - [Palliative total gastrectomy in advanced malignancies of the stomach]. AB - The diagnosis of the malignancies of the stomach is generally late (stages III and IV to a medium value of 80% of patients). Therefore it is necessary a surgical treatment in order to effectuate the extirpation of the lesions and to warn or to treat the complications in course of the neoplastic disease. The indications and the results of the palliative total gastrectomy (PTG) are analyzed in the present study. Between 1992 and 1999 there were studied retrospectively and prospectively during three successive periods of time, 217 patients (pts.) with PTG. This group of pts. represents 22.1% of the total number of pts. operated on for gastric malignancies (carcinoma particularly). The indication of PTG was established only after one complex evaluation of the every patient from the point of view of his general and biological status. 142 (65.4%/217) of the analyzed pts. presented at the admission in the hospital various complications of the malignant disease (gastric different stenosis, digestive hemorrhages, loco-regional invasion). The lymph nodes metastases and the systemic metastases (in the liver especially) as well as the invasion in the neighbouring viscera imposed sometimes the extension of the PTG with partial or total extirpation of the invaded organs. A proportion of 61% pts. were submitted to those type of enlarged surgical interventions without radical intentions; all the operated pts. remained by necessity in R1 or R2 types of operations. The reconstructive preferred anastomosis was effectuated with an Y jejunal ansa a la Roux completed or not by a "J" reservoir. Microscopically pathologic lesions were: adenocarcinoma and carcinoma (82%), followed by primitive lymphoma (11.5%), and others malignant forms of lesions (approximately 6%). The general perioperative morbidity was 32.5% divided in: 12.3% having a surgical cause (the majority anastomotic leakages +/- septically complications or precocious occlusions); 20.2% complications depending by the general status and altered biology of the patients. Perioperative mortality was 10.1% (22 pts.) comparable with other experiences published in the international literature in those forms of advanced gastric cancer operated by PTG. The average outcome of the operated followed pts. was 16-18 month (extremes 12 months and 29 months). A better quality of life and a variable disease-free period was obtained. As a conclusion we think that the PTG is a advantageous modality of surgical treatment with an acceptable rate of perioperative morbidity and mortality. The outcome of operated pts. is superior comparable with that of nonoperated pts. or with that secondary to other palliative surgical interventions. The importance of adjuvant treatment (chimeo- or radiotherapy) remain to be appreciated in the future. PMID- 12731151 TI - [Roux-en-Y gastrojejunostomy: indications, technique, results]. AB - 61 gastrojejunostomies were constructed between 1981-1999 in 58 cases (43 cases were operated between 1991-1999). 30 cases have previously suffered various gastric operations; in 28 cases the Roux-en-Y was the first gastric operation (27 operated on between 1991-1999). Radiologic and endobioptic studies were routine for diagnostic and follow-up evaluation, but various scintigraphic studies were performed in selected cases. The surgical technique was tailored for each case, according to the preoperative strategy. The indications for surgery were the following: 19-reflux disease; 14-severe peptic disease (9-reccurence, 3 postbulbar lesion, 2-juxtacardial ulcer); 8-peptic disease associated to reflux disease; 8-gastric cancer; 8-various early or late complications after gastrectomy; 1-lymphocytic gastritis. There were 4 early reoperations and 3 deceased. In 3 cases late reintervention was mandatory (for Zollinger-Ellison syndrome, peptic ulcer secondary to gastrectomy for cancer, Roux-stasis syndrome) and the Roux pattern was preserved; in 2 cases the antireflux effect of the operation was lost after the reoperation. PMID- 12731152 TI - [Particular clinical aspect of breast cancer]. AB - This scientific work intends to evaluate some of the particular clinical aspect of this cancer disease comparing the present medical tendencies with the clinical experience of the Department of Surgery I--Clinical Country Hospital Constantza. The scientific works was performed on a period of 5 years (March 1993-March 1998) on a number of 269 cases of breast cancer diagnosed and treated in our clinic. From the whole group of patients 33 particular cases were extracted: 26 were female patients aged over 70 years, 3 cases of bilateral breast cancer, 2 were breast cancer in association with other neoplasic locations and 2 male breast cancer. These cases were analysed using the: tumor location, clinical staging, hystopathological forms and complex therapy applied. This research work reveals the fact that breast cancer might present some particular aspect to be taken in consideration during the diagnosis and treatment of this frequent disease. PMID- 12731153 TI - [Follow-up evaluation of omphalocele treatment in children. The experience of the department of Pediatric Surgery and Orthopedics No.1 University Hospital Craiova]. AB - Abdominal wall malformations are a frequent category of congenital malformations in the last years in all Departments for Pediatric Surgery from our country. The authors are presenting a serie of 35 cases of newborns admitted and treated in the Pediatric Surgery and Orthopedics Department of the Clinical University Hospital No. 1 Craiova and in the same time they are evaluating in time the results after different surgical technics. From the point of view of the surgical techniques applied in the last 20 years the authors are communicating their results with the following procedures: Gross, Allen-Wren, Fufezan, and also their results with a nonsurgical procedure (Grob). The authors are manchenning that it is possible to combine (depending on the anatomo-clinical) the surgical approach with the non-operative management of the case. Clinical parameters such as sex, birthweight, immaturity, associated malformations, size of the abdominal deffect are the main element to predict the outcome of these patients and to choose the right management strategy, thus it is very important to investigate the pregnancy evolution and the birth period. The good results (82.9% surviving) are representing the prove of a good therapeutic decision being impossible to use synthetic materials for substitution (silicon, dacron, teflon, etc.) and in the same time an important guide for our future activity. PMID- 12731154 TI - [The main aspects of vesical risk in intraperitoneal surgery]. AB - They are described the main aspects of vezical risk in intraperitoneal surgery: subembilical celiostomy, haernios surgery, rectal surgery, gynecological surgery. It is shown few aspects of our experience in the treatment of haernias, vesicovaginal fistules secondary to total hysterectomy. PMID- 12731155 TI - [Gastric schwannoma ]. AB - The article presents the case study of a 78-year-old , female patient with a gastric schwannoma, with the symptoms of a superior digestive hemorrhage and secondary anemia. A surgical operation was performed and the patient was healed. The preoperative diagnosis carried out by means of echographic, endoscopic and computed tomography examinations was gastric tumour. Histopatological examination performed during the operation led to the final diagnosis of gastric schwannoma. PMID- 12731156 TI - [Thoracic esophageal neoplasm-therapeutical problems and post-surgery evolution]. AB - The authors are presenting a few considerations on Thoracic Esophageal Neoplasm, as resulted from an 18-cases study performed on patients that were operated between 1994-1999. The esophageal resection rate was of 50%, as follows: 7 Esophageal Resections and 2 Superior Polar Esogastric Resections. The digestive transit was reestablished by means of intrathoracic transposition of the stomach (6 cases) or of the right ileo-colon (2 cases). In one of the cases an Esogastric Anastomosis was performed at the neck level (cervical-right). Immediate post-op mortality after Esophageal Resection (1 case) was due to an acute respiratory distress syndrome (ARDS). The post-op complications were as follow: one anastomotic fistula associated with a purulent pleurisy, 4 non-infectious pulmonary complications and 2 cardiac complications (paroxysmal supraventricular tachycardia). The Discussions and Conclusions of the present work are presenting samples of surgical techniques, post-op complications and prognosis. PMID- 12731157 TI - [Late results after the Mark Belsey IV antireflux procedure in achalasia]. AB - Cardial achalasia (cardiospasm, megaesophagus) represents the failure or lack of relaxation capacity of the inferior esophageal sphincter; it is the second esophageal disease and is considered a premalignant lesion. Treatment of achalasia is surgical but palliative, considering the lack of an etiopathogenic therapy for this moment. We present a retrospective study done on 19 patients (10 males and 9 females, with a mean age of 49 years) operated in "Colentina" Surgical Clinic in the period of 1996-1999 for achalasia (presenting symptoms dysphagia, regurgitation, weight loss) by extramucosal esocardiomyotomy practiced on a length of 5-10 cm, by thoracic approach, followed by the Mark Belsey IV antireflux procedure. Results were good, without reflux pathology or dysphagia in the postoperative evolution. PMID- 12731158 TI - [Stapled anastomoses in colorectal surgery]. AB - The purpose of this paper was to analyze the advantages, indications and results of stapled circular anastomoses in colorectal surgery. In the last four years (1995-1998), fifteen patients underwent stapled anastomoses after Dixon's anterior rectal resection for cancers of upper and midrectum (11 patients), total colectomy with ileorectal anastomosis for malignant familial polyposis (1 patient), segmental colectomy of transverse and descending colon (1 patient with synchronous colic and rectal cancers, having concomitant rectal resection for cancer), previous Hartmann's resection for perforated upper rectal cancer (1 patient) or distal sigmoid diverticulitis (1 patient). The anastomoses have been performed in end-to-end fashion (11 patients), according to the Knight's technique (2 cases) or in side-to-end fashion (2 patients). As most frequent associated technique with stapled anastomoses, anterior rectal resection for cancer was performed with 2 cm of clearance beyond the macroscopic margin of tumor. Distal margin of resection was histologically verified and it proved to be free of tumor cells. There was no operative mortality. Anastomotic leakage occurred in three patients because of imperfection of stapled anastomosis (2 cases) or after local irradiation (1 case). Spontaneous closure was seen in one patients. The other two patients needed reoperation for suture or colostomy. Late clinical, endoscopic and X-ray controls did not discover local recurrences. Functional results were good in terms of stool frequency and continence. In conclusion, stapled fashioned anastomoses have the main indication in sphincter saving Dixon's and Hartmann's procedures. In these cases, stapled anastomoses are easier than manual technique, reduce operative time and improve suture reliability. PMID- 12731159 TI - [Mechanical suture in colorectal surgery]. AB - THE OBJECT: Of this work was the study of using, as well as the utility of the mechanical sutures in colorectal surgery; because of the special caution needed to be taken for any colonic or rectal suture, more than any other digestive segment. The frequency of the postoperative fistulas after the suture and anastomosis is higher at this level and so it increases the period and costs of the hospitalization. MATERIAL AND METHOD: We studied the possibilities of performing and evolution of 64 mechanical sutures for 19 patients, with colorectal pathology, hospitalized in our department from july 1999 to december 2000. RESULTS: We performed 64 mechanical sutures, as followed: 47 in open surgery and 17 in laparoscopic. From all these, 56 was bowel sutures, 8 of them were vascular (in laparoscopic, for cutting the most important vascular pedicles). We did 18 anastomosis: 15 in open and 3 in laparoscopic surgery. It was 2 postoperative fistulas from all 56 intestinal sutures (3.57%). We haven't any intra or postoperative bleeding from the vascular anastomosis. It was 3 intraoperative bleeding from the intestinal anastomosis, and only 1 case of postoperative bleeding (5.26% of the cases: 1.56% of all mechanical sutures). In only one case, the mechanical suture couldn't be initially done, but it succeeded after the removing of the segment of the bowel involved. CONCLUSIONS: Mechanical sutures offers a high level of safety to the colorectal anastomosis. It provides a very good vascularization to the anastomosis and decreases the time needed for performing the suture or anastomosis, versus manual sature. Also, for the patients with rectal ampular neoplasm, it creates the possibility of anal sphincter preservation by making a low colorectal anastomoses--which is difficult by manual suture. PMID- 12731160 TI - [The use of Sono-Surgery in breast cancer surgery]. AB - THE AIM: Of this research is presenting the using of Sono-surg as a technic possibility in breast cancer surgery. MATERIAL AND METHODS: Are presented and analyzed cases of mastectomy and laparoscopic oophorectomy in which since 1999, was used like element of hemostasis Sono-surg. RESULTS: In 23 cases of laparoscopic oophorectomy using Sono-surg, was reduce the time of intervention, incidence or usually complications. The idea of using Sono-surg in mastectomy too, make sure a perfect hemostasis even if the time of the intervention is longer. CONCLUSION: Introducing Sono-surg in surgical treatment of breast cancer is a therapeutic solution which may be used with postoperative profits. PMID- 12731161 TI - [Laparoscopic splenectomy--lessons learned from a series of 40 cases. The advantages of the postero-lateral approach]. AB - Until October 2000 in our surgical department 40 patients underwent laparoscopic splenectomy. Seven patients were converted to laparotomy for completion of splenectomy. All of these occurred in the first 20 patients and were due to bleeding. There was no postoperative mortality. Postoperative complications occurred in two patients: one abdominal wall hematoma and a postoperative hernia. Indications were idiopathic thrombocytopenic purpura, hereditary spheocytosis, autoimmune hemolytic anemia and others. In the last 30 cases we used the postero lateral approach, that results in reduced blood loss, fewer patients that needed to be converted to open surgery and shorter postoperative stay. Accessory spleens can be successfully localized and with a carefull technique the capsular fractures can be minimized reducing the risk of splenosis. PMID- 12731162 TI - [Impact of surgery training upon the results of inguinal hernia treatment operated using laparoscopy (757 cases)]. PMID- 12731163 TI - [Strategy and tactic in the treatment of local advanced rectal cancer]. AB - In local advanced rectal cancer (LARC) was defined at the work group in rectal cancer as a tumour what invade the serosa or neighbouring organs, associated with invaded perirectal or mezorectal nodes, with internal fistulae, peritoneal carcinomatosis and locoregional recidives. On a trial of 97 patients, the authors present personal experience, in comparison with literature data regarding therapeutic strategy and tactic of parameters: operability and the moment of operation, indication of preoperative radiotherapy, the type of operation, excision of the metastasis, adjuvant therapy and attitude of locoregional recidives. The survival of the patients was 21.6% at 3 years and 15.4% at 5 years. PMID- 12731164 TI - [Injuries to the duodenum and pancreas in 42 operated cases]. AB - 42 consecutive patients who sustained injuries to the duodenum or/and pancreas were admitted to our hospital. Over a twenty year period 32 blunt injuries and 10 penetrating injuries were encountered. Penetrating injuries were always suspected and treated by time: following blunt injury diagnostic delay was encountered in 14 patients and insufficient surgical procedure because of intra-operative misinterpretation in 2 patients. Most of the patients had associated intra abdominal organ injuries. Adjusts to diagnosis such as abdominal roentgenograms, serum amylase levels and gastroduodenography was not helpful. CT scan and ultrasound allowed to confirm the suspected diagnosis in 3 cases only. Intraoperative diagnosis was also challenging. Complete mobilization of the strictures surrounding the duodenum and the pancreas to provide entire exposure was necessary in 12 patients treated first in a peripheral hospital, diagnosis of the injury have been missed at first laparotomy and reoperation was necessary in all of them. Suture closure of the duodenum and drainage of the pancreatic region wee the most common reparative techniques used. More complicated procedures with pancreatic and/or duodenal resection were performed in 12 patients. Overall mortality in patients surviving more than 24 hours was 14%. Suture live dehiscence after delayed operation (4) and 2 deaths due to brain injury. PMID- 12731165 TI - [457 inguinal and femoral hernioplasties with synthetic prosthetic materials]. AB - The authors retrospectively analyse 457 inguinal and femural hernioplasties with synthetic prosthesis materials, during a period of 10 years. There have been discussions going on about the prosthesis utilisation indications (advanced stages, complex inguinal hernia), type of prosthesis that has been used, the modalities of inserting a prosthesis and the surgical techniques employed. In 164 (34%) of the operations the mesh has been used as an reinforcement and 293 (64%) of the interventions were "tension-free" operations. The average reccurrence was approximately 1%, during a period ranging between 1 to 5 years. PMID- 12731167 TI - [Diagnosis of textilomas]. AB - Despite all the precautions during surgery, it may be assert that obliviousness of a compress or of a mesh remains still possible. In order to define this accident a non-medical term, textiloma, is used, only in hospital environment. Radiology and medical imaging represent the main tool for the diagnosis of textilomas. Plain film of the abdomen and/or ecography may be sufficient for revealing the textiloma. Just in a few cases a computed tomography may be required. In order to demonstrate all these, using two cases, we'll emphasize the main radio-semiological aspect of textilomas, and thus intending to establish an examination algorithm. PMID- 12731166 TI - [Post-operative temporary enteral nutrition methods]. AB - THE AIM: Of the present work is to promote duodenostomy as the postoperative enteral nutrition way, when surgery ends in an anastomosis in which one of the partners is the esophagus (esophagectomies, esogastrectomies and total gastrectomies). MATERIAL AND METHOD: Consisted of 45 cases 815 total gastrectomy, 14 esogastrectomy, 6 esophageal resections and 10-esophageal plasty) in which we used: nasofaringoesogastric or nasofaringoesojejunal probes (14 cases); Witzel jejunostomy (11 cases); gastrostomy (10 cases); duodenostomy (10 cases). THE CONCLUSIONS: Show the many advantages of duodenostomy as compared to other enternal nutrition methods: technical simplicity, patient's comfort, avoidance of complications involved by the use of nasopharingoesodigestive probe or by jejunostomy, etc. PMID- 12731168 TI - ["Spontaneous", "delayed" and "occult" ruptures of the normal and pathologic spleen. Nosologic classification]. AB - The anatomic particularities and the diversity of the splenic pathology determine a wide spectrum of the traumatic lesions of the spleen. The classical acute rupture with consecutive hemoperitoneum and mandatory surgical indication is opposed to the controversial "spontaneous" ruptures of the normal and pathologic spleen and also to the delayed and occult ruptures the lasts of them sometimes minimal or with chronic evolution. A series of eight cases all males with ages between 40-77 years is presented in order to exemplify all these entities. There are underlined the variety of etiologic conditions, the difficulties of the diagnosis which impose an insistent anamnesis and clinical examination (searching even a "trivial" trauma) completed with ultrasonography and CT scan which are preferred to radionuclide scan of arteriography and finally peritoneal lavage after punction, laparoscopy and even laparotomy. Splenectomy--often laborious- was effective in all our patients (obviously only temporary for the cases with nonhodgkin malignant lymphoma and respectively with splenic metastasis from a renal carcinoma). In some situations--especially in children--the conservative treatment can be considered. PMID- 12731169 TI - [Exceptional etiologies in upper digestive tract bleeding ]. AB - 7 cases, considered as being true etiological exceptions selected from 756 upper gastrointestinal bleeding, are presented. The causes of bleeding were: pancreatic pseudocyst with intracystic hemorrhage broken into duodenum (2 cases), the nonepithelial gastroduodenal tumor (3 cases), the aneurysm of gastroduodenal artery broken into duodenum (1 case) and the aortoduodenal fistula in one patient with a bilateral aorto-iliac by-pass (1 case). The etiological diagnosis could not be established in any cases before the operations. All the cases were operated on, the operation being imposed by the severity of bleeding and having the haemostasis as a main purpose. PMID- 12731170 TI - [Acute abdomen and diabetic patients--difficulties of diagnosis and therapeutical decision]. AB - We present a retrospective study based on 50 diabetic patients with acute abdominal diseases. Usually, clinical features were not typical, without defining signs of acute abdomen, despite frequent severe anatomo-pathological forms (6 of 12 acute appendicitis were gangrenous, with generalised or localised peritonitis; 15 of 22 acute colecystitis were gangrenous). In diabetic patients, with metabolic disorders and cetoacidosis, positive diagnosis and the decision of laparotomy are difficult problems, often delated, with a negative influence on the evolutions and prognosis of these patients. PMID- 12731171 TI - [Intraoperative ultrasonography, dissection with ultra-shears and trans-cystic choledochoscopy during laparoscopic approach in obstructive lithiasis icterus case]. AB - The authors present the case of a 48 years old male with lithiasis obstructive icterus who was operated by laparoscopic transcystic approach with choledochoscopy. This minimally invasive technique was very spectacular trough the using of laparoscopic ultrasonography, dissection with Ultra-Shears device and transcystic choledochoscopy. Careful watching of abdominal laparoscopic view and transcystic choledochoscopic view is the particularity of laparoscopic transcystic management of choledocholithiasis. This metod don't let us to explore the biliary tract before the junction between common bile duct and cystic duct. PMID- 12731172 TI - [Retrograde cold knife endo-pyelotomy in ureteropelvic junction stenosis]. AB - Between September 1998 and July 2000 we performed a new procedure of ureteropelvic junction (UPJ) stenosis treatment: retrograde endopyelotomy (REP). So, 17 patients (7 males and 10 females) between 23 and 56 years old with hydronephrosis (IInd grade--3 cases, IIIrd grade--12 cases and IVth grade--2 cases) were evaluated with an unitary protocol: gray-scale and duplex Doppler (resistive index--RI measurement), IVP, ureteropyelography and renal scintigraphy in selected cases. The hydronephrosis was primitive in 12 cases and secondary in 5 cases (after previous open surgery). We excluded from our series of patients cases with aberrant vessels (polar inferior), associated renal stones, important hydronephrosis and young patients. Video assisted endoincision was realized at 6 and 3 o'clock position on the left side and at 6 and 9 o'clock position on the right side with Wolf semirigid ureteroscopes, Storz special knife/scissor. Double J indwelling catheter (8 Fr.) was removed after 6 weeks. The evaluation has been made at 3.6, 9 and 12 months demonstrating normal pyelocaliceal system in 10 cases (58.8%), a significative reduction of hydronephrosis grade in 3 cases (17.6%), mean RI = 0.63, with the presence in 15 cases (88.2%) of RI under 0.70. Stable good results were found in 14 cases (82.3%). After the failure of this technique (stenosis in 2 cases) the REP was practiced again. We didn't find intraoperative complications. Our conclusions despite the relative short follow up period are very much in favor of this modality of treatment, minimally invasive, being an efficient technique in the modern therapy of hydronephrosis. PMID- 12731173 TI - [The experience of our clinic in the treatment of vesicovaginal fistulas]. AB - In this work paper we have presented the experience of our clinic in the treatment of vasicovaginal fistulas (17 cases), secondary surgical procedures. The frequency of vesicovaginal fistulas is very high after gynecological surgery (14 after total hysterectomy for uterine cancer, uterine fibroma or uterine necrosis secondary to septic abortion). In all cases authors used Diettel-Forgue Legueu procedure, with good results in 88.30% of cases. PMID- 12731175 TI - [Results of clinical treatment in gastric neoplasm]. PMID- 12731174 TI - [Experimental model of heterotopic renal transplantation in rat]. AB - This study describes the experience with a new technique for heterotopic kidney transplantation in the laboratory rat. This technique combines already known aspects in renal experimental transplantation, with technical improvements added to each surgical step. Ten heterotopic kidney transplantation were performed. Immediate postoperative survival was 100% and at 48 hours was 90%. The patency of the vascular anastomosis checked at 72 hours in survivors was 100%. As a preliminary study, 20 renal hilum dissections were done, noting the topographical aspects of the renal vessels. Representing a standardized and easy to use model, this solid organ transplantation technique is a valuable tool for studying the immunological and physiopathological aspects of transplantation and for refining the microsurgical skills. PMID- 12731176 TI - [The systematization and the etiopathogenicity of diaphragmatic hernias]. AB - The author, based on up to date published dates, intends to present the classification and ethiopathogeny of the diaphragmatic hernias, except the aesophagic hiatus oms. This is an interesting chapter of the borderline surgery (abdominal and thorax). They are placed on the second position in frequency (after the hiatal hernias) in the diaphragmatic pathology; they are internal hernias, through congenital or obtained holes which allow to abdominal viscera to pass into thorax. They are--in the most cases, even elderly ones-congenital, result of the abnormalities in the embrionary growth of the diaphragm. A special place' is represented by the traumatic hernias. PMID- 12731177 TI - [The useless appendectomy]. AB - The appendectomy is still the most frequent operation in general surgery, being after performed without complete investigation. Because the appendix, through its lymphoid tissue with implication in the immunoglobuline secretion, has an important role in immunitary system criteria were searched to justify the appendectomy and to reduce the number of useless appendectomies, the proportion of the normal appendices being around 15%. Instead the basic triad-fever over 38 degrees C, parietal reaction and leucocitosis over 10.000/ml--measuring of reactive protein C, echography, CT, laparoscopy and antileucocite antibody labelled with TC99, would contribute to a better selection of patients for appendectomy. Surgeon's responsibility regarding the technique and the moment of operation, oblige him to take the proper decision. PMID- 12731178 TI - [Difficult laparoscopic cholecystectomy]. AB - From 2505 cases operated between 1994-2000, the authors selected a lot of 889 cases of difficult cholecystectomy, which after classical criteria would have been classified as contraindications of laparoscopic procedure. This study analyzed of intraoperative incidents, the conversion rate and particularities of operative tactics and techniques. The lot of patients includes 889 cases of difficult laparoscopic cholecystectomy. The patients were selected according to classical contraindications of laparoscopic cholecystectomy: obesity, liver cirrhosis, previous operated abdomen, severe cardio-pulmonary diseases, ages over 70 years, acute cholecistitis. The rate of intraoperative incidents and accidents was 19.9% (177 cases), in comparison to intraoperative incidents and accidents rate in "easy" laparoscopic chole-cystectomy which was 15.5% (251 cases from 1616 patients). The conversion rate in case of difficult laparoscopic cholecystectomy was 6.1% (55 patients) and 3.2% in the lot of "easy" cholecystectomy. The postoperative morbidity was 6.4% (57 patients) and 5.5% (147 patients) in the lot of "easy cholecystectomy. The postoperative mortality was 0.6% (6 deaths) and 0.3% (5 deaths) in the lot of "easy" cholecystectomy. In this paper are discussed the modalities and the technical particularities of a difficult laparoscopic cholecystectomy, able to lowering the risk of operative incidents and postoperative complications. In conclusion, laparoscopic cholecystectomy can be safely performed to the majority of patients with lithiasic cholecistitis, by an experienced surgical team. PMID- 12731179 TI - [Pancreatic insulinomas]. AB - In the period of 1980-1999 (20 years) were operated 18 patients (8 males and 10 females) in "Colentina" Surgical Clinic, the average age being of 45 years, with hystopathologically confirmed insulinomas. The topography of these lesions was in the head of the pancreas (4), in the isthmus (5), in the body and tail (9). The classical clinical and biochemical signs were presented in 13 cases, the rest of the patients presenting atypical signs of disease. These patients had benefit from the surgical treatment. The approach was bi-subcostal in 11 cases and by upper and lower midline incision in the rest of cases. We practiced enucleation in 5 cases, corporeal-caudal pancreatico-splenectomy in 10 cases and cephalic pancreatico-duodenectomy in 3 cases. The malignancy of the tumour was hystopathologically confirmed in 30% of the situations. The postoperative mortality was in 3 cases, by acute pancreatitis, MSOF and myocardial infarction. We consider that in case of these generally benign tumours the most efficient therapy is the surgical one. PMID- 12731180 TI - [The anal incontinence-- study on 20 operated cases]. AB - The authors present 20 cases operated for anal incontinence. Two techniques were performed: direct repair (18 cases) and Musset-Cottrell procedure (2 cases). The results were excellent in 12 cases, good in 5 cases and satisfactory in 3 cases. The method of choice seems to be the direct repair of the anal sphincter after a proper local and general preparation. PMID- 12731181 TI - [Hyperfunctional parathyroid carcinoma]. AB - The parathyroid cancer is mentioned in literature with an incidence of 0.5-5% in the etiology of the primary hyperparathyroidism. The authors present the case of a 45 year old female with diagnosis of "primary hyperparathyroidism" based on clinical, ultrasonographic and biochemical investigations. A right superior adenoma of 3 x 2 x 1 cm (150 mg) has been found and extirpated. The histological examination showed an adenoma with predominant "chief" cells. Three years after surgery the patient had a recurrence of the clinical and hypercalcemie syndrome (with more severe damages). The surgical reexploration showed the right thyroid lobe with a nodular aspect and in its inferior pole a enlarged parathyroid gland of one cm size was found. An en-block exeresis including the right thyroid lobe with isthmus as well as the mentioned lesion and the half of left superior parathyroid, the left inferior one together with retrosternal fat tissue have been performed. Frozen sections completed by paraffin examination established the diagnosis of hyperfunctioning parathyroid carcinoma. We have emphasize the issues that could suggest the initially preoperative true diagnosis: the large size of excised adenoma and the relatively quick recurrence of the phenomena of parathyroid hyperfunction. The surgical principles and strategies in the treatment of parathyroid cancer are also discussed. PMID- 12731183 TI - [Unusual hemorrhagic complication after surgery for severe generalized appendicular peritonitis]. AB - The 22 year old male patient, was admitted in hospital for severe generalized peritonitis subsequent to acute perforated appendicitis and toxico-septic shock. On examination and relying on the previous history of the patient onset of the perforation was start assessed to have occurred some days earlier. Severe generalized, putrid peritonitis was found on surgery of the peritoneal cavity. Appendectomy was successfully carried out with a simple ligature of the appendix stump, and the ligature of the mezooappendix was performed without identifying the appendicular artery. Early in the postoperative stage there occurred diffuse bleeding localized in the peritoneal cavity. The source of the bleeding could not by found at the first reoperation. Subsequently the septic syndrome evolved simultaneously with the bleeding in a milder form, however, leading to growth in size of retroperitoneal hematoma. On marking the diagnosis, relying on CT examination, a new, second surgery was performed which afforded evacuation and drainage of the retroperitoneal hematoma. The authors have remarked and have tried to clear up the circumstances which had been conductive to the occurrence of hemorrhage, a thing absolutely unusual in the evolution of diffuse peritonitis by perforated acute appendicitis. PMID- 12731182 TI - [Recurrent thyroid neoplasm with Hurtle cells-- case report]. AB - The thiroidian tumor with Hurtle cells represents a rare clinical form of thyroid neoplasia and has a particular biological behavior. Tumors included in this category are those in which more than half of the cell population is made up of Hurtle cells. We present the case of a 58 years old woman with a recidivant oncocitic adenoma, four years after a subtotal bilateral thyroidectomy. The pathological examination using HE, van Gieson, AgNOR stains, the positive imunoreaction for thyreoglobulin LSAB and the ultrastructural examination established the definitive diagnosis. The surgical treatment is the main therapeutic measure and must be performed under oncological curative principles: lobectomy for the benign forms and total thyroidectomy for the malignant ones. The malignant potential is higher than for other well differentiated thyroidian carcinomas. PMID- 12731184 TI - [Comparative study about alloplastic surgical procedures of incisional hernias]. AB - This retrospective study analyzes a series of 604 patients with incisional hernias, operated on between 1994 and 1998. The hernias where divided according to their dimensions and their reccurrence. The postoperative outcome and follow up leaded to some conclusions: although the postoperative immediate complications were frequent after the alloplastic procedures, the long term results, judged by the rate of reccurrence and the rejection of the mesh, were better. With the alloplastic procedures the best results were obtained when the mesh was fixed in a retromuscular--properitoneal position. PMID- 12731185 TI - [Laparoscopic surgery of inguinal hernia (personal technique)]. AB - The author shows a personal, original, quick, cheap and simple videoscopic procedure that is also nonaggresive and without recurrences. The method updates the REVERDIN needle features. PMID- 12731186 TI - [Analysis of various gastric carcinoma classification systems, through the prism of a study performed at Craiova IV surgery clinic]. AB - INTRODUCTION: The aim of this work is to analyze the various gastric carcinoma classification systems, trying, at the same time, to establish connections between the anatomopathologic evolutionary and prognostic aspects. CASE MATERIAL AND METHODS: The study was performed on a series of 66 patients who undergone excision surgery for gastric carcinoma. The study material are 31 cases of gastric cancer diagnosed in Clinical Hospital CFR Craiova, and another 35 cases diagnosed at the "I.V. Babes" Pathology and Medical Genetics Institute in Bucharest. The excised pieces underwent anatomopathological investigation using histopathological, histochemical and immunohistochemical methods. RESULTS: From the macroscopic point of view, the most common tumor pattern was Bormann 2 ulcerovegetative, targeting patients over 60 years of age. In what the invasion level is concerned, most carcinomas were sanctioned at the time when the tumor was invading the serous fluid (pT3) or even the perigastric tissue (pT4). The microscopic criteria was approached by us using Goseky's method, the most common being Goseky stages 2 and 4, in equal rations. The distribution of cases depending on the OMS differentiation degree, shows a preeminence of low differentiated carcinomas (G3). CONCLUSIONS: The abundance of classification methods in gastric cancer shows the complexity of the matter but none of the classifications is perfect. PMID- 12731187 TI - [Pseudomyxoma peritonei (gelatinous peritonitis )]. AB - Pseudomyxoma peritonei (P.P.) is characterised by the presence in the peritoneal cavity of 3 elements--mucinous neoplasic cells, mucinous ascites and diffuses mucinous implants. We analysed the patients treated in our service and we made a literature review. The study is a retrospective analysis on the 1984-2000 period. 6 patients were diagnosed with P.P., 2 females and 4 men, with medial age 63 years (53-75). Clinical findings and biologic analysis were not specific and the imagistic explorers didn't permit the preoperative diagnosis. The patients were submitted to variate surgical procedures, based on the benign (4 cases) or malignant (2 cases) character of the disease and on the origin of the lesions: cystadenoma of the appendix with secondary tumours of the ovary (the 2 females) and, respectively, cystadenoma and cytsadenocarcinoma of the appendix, mucinous paraenteric cyst with pseudomyxoma retroperitonei, mucinous recto-sigmoidian neoplasm (the 4 men). We practiced intraperitoneal chemotherapy with Thio-Tepa in 5 cases (intraoperative in 4 cases) and systemic, with 5-FU and mytomicine, in one case. The operative morbidity and mortality were null. All the patients were follow-up (average 4.2 years). The global 5-year survival rate was 60% (100% in the benign disease and 0% in the malignant form). CONCLUSION: P.P. is a rare disease, still poorly understood. The origin of the disease is the appendix (70 80%) and less frequently the ovary. It was divided in two distinct forms: disseminated peritoneal adenomucinosis and peritoneal mucinous carcinomatosis. The treatment associates radical surgery and intraperitoneal chemohyperthermia, in specialised centres, but the prognosis still remains poor (50-70% 5-year global survival rate). PMID- 12731189 TI - [Laparoscopic reoperations]. AB - The progress of laparoscopic techniques have made possible video-assisted miniinvasive surgery in patients with indication of reoperation. Knowing that there are many controversies against this type of surgery, the authors tried to formulate their own point of view in this matter by analyzing their experience in such particular cases. In this study there are included both laparoscopic re operations (or re-laparoscopies), consecutive to video-assisted surgery, and the mini-invasive procedures performed for complications after open surgery. Of the total 3901 laparoscopic operations (in 3714 patients) we performed 34 laparoscopic re-operations for postoperative complications occurred in patients previously operated on in our clinic or in other departments (29 re-laparoscopies and 5 after open surgery) The objective of the study was to verify if the laparoscopic techniques are useful in diagnostic and treatment of the postoperative complications which need surgical re-exploration, and the conclusions have shown that laparoscopy may be appropriate in re-exploration of the surgical patients, if the rules of the operative management is respected and the well defined standards are fulfilled. PMID- 12731188 TI - [Liver transplantation--considerations over 8 cases operated in the year 2000]. AB - In the year 2000, at the Department for General Surgery and Liver Transplantation from The Fundeni Clinical Institute Bucharest, seven OLTs and one living-related transplantation were performed in 6 adults and 2 children. Postoperative complications were: bile leakage, hemoperitoneum, lower gastrointestinal hemorrhage, parietoabdominal hematoma. There was only one postoperative death due to septic complications in the 18th p.o.d. and one late death due to pneumonia of unknown origin. After the results in the year 2000 there was an increased number of donors and referrals. We consider that now in Romania this is an established program that will continue depending on the number of donors and financing. PMID- 12731190 TI - [Therapeutical options in lithiasic biliary fistula]. AB - Chronic lythiasic cholecystitis is a disease distinguished by the pathologic changes because of the chronic inflammation of the biliary extrahepatic tree. Sometimes these morphological changes are associated with internal biliary fistulas arising spontaneously in patients with advanced calculus cholecystitis. The vast majority of fistulas result from the adherence of the inflamed gallbladder or common bile duct to an adjacent viscus and erosion of the gallstones into the adherent organ. The authors analyze an amount of 43 patients with bilio-biliary and bilio-digestive lythiasic fistulas, caused by the long evolution of chronic lythiasic atrophic cholecystitis, for 126 cases which were operated in the Surgery Department of the Caritas Clinic Hospital along 20 years. In these 126 cases the surgical strategy was determinated by the method of dealing with the pericolecystitis sclerotic blocks, by the identification methods of the biliary elements and by the approach of the main billiary way and digestive loops injuries occurred after suppressing the fistulae. The surgical solutions adapted for each separate case and the advantages of the axial drainage of prostheting the surgical reconstructions of the main biliary way as well as the results obtained along this project, are in fact the aims of this presentation. PMID- 12731191 TI - [Surgical treatment of duodenal ulcer]. AB - The medical and endoscopic treatment of duodenal ulcer are decreasing the frequency of surgical treatment in this disease. The authors study the operations performed for duodenal ulcer within the period 1989-1999 in the County Hospital Baia Mare. The decrease of the rate of surgical interventions is the pure effect of the medical treatment, as long as the endoscopic treatment is not yet available in our service. The rate of ulcer--induced perforations remained, however, unmodified (48% of total operatory indications), as well as the postoperative morbidity and--mortality (18% respectively 9%). The last category seems not to be influenced by the type of chosen surgical procedure, but by the patient's age, duration of the disease, and associated pathology. PMID- 12731192 TI - [The intraoperative colonic irrigation in emergency surgery]. AB - Bowel preparations is frequently impossible in various ante colonic diseases, such as left-sided colonic obstruction. The goal of intraoperative colonic irrigation is to obtain, during surgery, a bowel preparation offering the possibility of primary resection with immediate anastomosis, when preoperative bowel preparation has not been feasible. Technical aspects of intra-operative colonic irrigation are described. Indications for this methods are presented: left-sided obstructing carcinomas, diverticulitis, more rarely inflammatory stenosis or functional obstruction. The surgical management of left colonic emergencies has evolved in the past few decades. Recently, there has been increasing interest in resection with primary anastomosis in selected cases. The post operative mortality rate was 13 per cent. The incidence of clinical anastomotic leakage was 6.65 per cent. PMID- 12731194 TI - [Merkel cell carcinoma]. AB - Merkel Cell Carcinoma (MCC) is a rare and aggressive neuroendocrine dermal neoplasm. This study is a retrospective outcomes analysis of two cases of MCC with data regarding clinical, histopathological, immunohistochemistry and also surgical, chimio and radiological treatment. MCC is a rare dermal tumors, this tumors are most predictable found on sunexposed sites. Diagnosis is best accomplished by a thorough clinical evaluation coupled with light microscopy and defined panel of immunohistochemical studies which are necessary for the definitive diagnosis of Merkel cell carcinoma (cytokeratins, neuron specific enoiase and chromogranin). A lot of other disease must be included in the differential diagnosis. MCC is an aggressive tumor with local or locoregionale extension and distant spread by hematogen or lymphatic way. Surgical excision of tumor and regional lymphadenectomy is the first step of treatment completed with radiotherapy and chemotherapy bat in advanced studies the rate of local or distant recidives is high. PMID- 12731193 TI - [Abdominal hemangiopericytomas]. AB - We present two cases of hemangiopericytoma, which are analyzed according to medical literature, although only few cases have been reported. Clinical, imagistic and histological findings of these rare vascular tumors are discussed. PMID- 12731195 TI - [Transsacral fistula of an ileal pouch-anal anastomosis]. AB - For patients with ulcerative colitis and familial adenomatous polyposis the restorative proctocolectomy with ileo-anal-pouch anastomosis is the surgical treatment of choice. Leakage from the ileo-pouch anastomosis is the surgical treatment of choice. Leakage from the ileo-pouch-anastomosis can be a difficult to manage complication, which in some cases resists all attempts at local repair. A surprising complication of a 28 years old woman patient with an ileo-anal-pouch anastomotic fistula is presented. The fistula developed the 4th day postoperatively. Local irrigation and transanal drainage seemed to have a good result, the patient being examined after two weeks. During an apparently better evolution, after one month, the patient developed a transsacral fistula with local abscess and osteolysis. The ileo-anal-pouch anastomosis was converted to a less comfortable conventional ileostomy, but with good local and general final result. PMID- 12731196 TI - [Endoscopic sphincterotomy in the treatment of common bile duct benign lesions (experience with 100 cases)]. AB - The authors present the experience of the first 100 endoscopic sphincterotomy performed in Caritas Hospital, Surgery Department between 1997 and 2000 for CBD benign lesions treatment. The patients to whom we performed this procedure had a biliary syndrome associated mostly to obstructive jaundice of unknown etiology previously diagnosed echographically or by ERCP. The etiology was in 72% of the cases represented by migrated, residual or primary common bile duct stones. The aim of the procedure was the removal of the stones. We performed endoscopic sphincterotomy either before or after laparoscopic or open surgery. The morbidity was 8% and the related mortality was 0%. Endoscopic sphincterotomy is an efficient procedure for the treatment of the benign obstruction of the common bile duct and not only. Our results were similar to those published by other authors. PMID- 12731197 TI - [ A new technique for the treatment of varicose veins]. AB - The thoroughgoing study of the venous wall and venous flow using fundamental research and diagnostic techniques allowed us to apply new therapeutic methods in the primary varicose veins of the lower extremities (PVV). The aim of the present study is to present a relatively new method of conservative surgical treatment which combine the external valvuloplasty technique with the technique proposed by Franceschi (TF). MATERIAL AND METHOD: Between 1992-2000 we applied on 25 patients with PVV Mach II, the technique of external muffing of the long saphenous vein from its junction with the common femoral vein retrograde for 6-8 cm, for a venous diameter of 0.5 cm interrupting at the same time the long or the short ebbing (clinical and Doppler criterions). In 5 patients we performed the valvuloplasty using TF. The patients were annually evaluated; in the present we have a group of 10 patients up to 8 years after surgery. RESULTS AND CONCLUSIONS: 1. Conservative methods of external valvuloplasty associate with Franceschi technique have a solid physiopathologic and pathogenic basement. 2. The materials used for the external muffing (Dacron, PTTF, polyamide net) are very well tolerate. 3. Relapses were present in 8 years at 30% and in 5 years at 10% of the patients who undergo the operation. PMID- 12731198 TI - [Breast cancer-estrogens relationship]. AB - Variations of estrogen (produced mostly in the ovaries) are under enzymatic influence of aromatases, acting also as oncogenes through the cytochrome CYP17 and CYP19. Their degradation trough methoxylation, resulting in both carcinogenetic and protector compounds, is influenced by 2, 4 and 16 +/- hydrolases. Tissue sensibility to oestrogen depends on the amount of receptors and the plasma level of oestrogen. They are regulating cell's growth and differentiation, depending on subject's age. Paraclinic markers such as the amount of regulating receptors, plasma oestrogen level, breast density and bone density are elements in evaluating breast cancer risk. Among the risk factors of occurrence and development of breast cancer were cited also early menarche, late term pregnancy, late menopause, postmenopausal obesity, smoking and the diet rich in fats and fatty acids, alcohol and antioxidant vitamins. PMID- 12731199 TI - [Our experience regarding a series of adrenal surgical diseases]. AB - AIMS: The problems of positive and differential clinic-pathologic diagnosis, the optimal therapeutic decision and importance of surgery in a series of patients with adrenal gland (AG) diseases was studied. MATERIAL AND METHODS: Between 1987 1999, 91 patients (pts.) with adrenal diseases were operated on. Sex ratio was 63 f/28 m and the medium age 42.3 years (extremes 14 and respectively 75 years old). The lesions were localized in medullar, cortical and in the both components of AG. 1) The cortical determinations were represented by 40 (44%) pts. with: a) Cushing's syndrome 37 pts. histologically grouped: 24 pts. (63%/37) with benign, diffuse hyperplasia of cortico-adrenal glands; 4 pts. with cortical benign adenoma; 7 adenocarcinoma; 2 with borderline lesions. b) Conn syndrome--3 pts. 2) Medullary lesions (Phaeochromocytoma)--30 pts. (33%/91) with following microscopically lesions: 22 typical phaeochromocytoma; 3 benign adenoma; 5 carcinoma and 1 medullo-adrenal paraganglioma. One patient have had bilateral medullo-adrenal lesions: left carcinoma and right benign adenoma). 3) Non functioning tumors (Incidentaloma)--21 pts. (23%/91) (17 malignant and 5 benign). The distribution of the whole group of 91 pts. was: 62 with benign lesions (diffuse hyperplasia or tumors), 26 malignant tumors, 1 both of them and 2 borderline lesions. The diagnosis was the result of clinically, biochemical hormonal tests and imagery examinations (computed tomography especially) with postoperative pathological confirmation. The surgery applied was:--bilateral adrenalectomy 16 pts.;--right adrenalectomy 20 pts.;--left 26 pts. (two through laparoscopic approach);--tumor exeresis 26 and--3 biopsy of the tumors. The dimensions of the operatory specimens were: < 10 cm--67 pts.; 10-20 cm--16 pts.; > 20 cm--2 pts.; undetermined 6 pts. For malignant lesions the main parameters (in various associations) were: neighborhood invasion--18 pts.; distant metastasis--5; metastatic lymphnodes--13; the microscopic grading (G1-1; G2-9; G3 7 pts.). Three cases imposed iterative interventions for a recurring adenoma of the cortically AG or for restant tissue in Cushing's syndrome. There were necessary 28 supplementary operative (partially or totally) organs exeresis. All pts. with malignant lesions were postoperative treated with chemio- or/and radiotherapy. RESULTS: Postoperative morbidity: 18 (20%) pts.; depending of the surgery 8 pts., secondary of the patient general status 12. General postoperative mortality 4 (4.8%) pts. determined by severe cardio-vascular complications. CONCLUSIONS: 1--The surgical diseases of adrenal glands are difficult to diagnose and are based primarily on the clinical information's and confirmed by the hormonal and imagery examinations (errors are possible). 2--Operatory indications, especially for hyperplasic bilateral forms in Cushing's syndrome (one step or two steps surgery) must be very well documented and carefully established. 3--There are cases which impose recurrent operations. 4--A correct operative indication and technical surgery procedure are followed by good results. 5--The laparoscopic approach of the surgical lesions of the AG is a good alternative for the open surgical approach with the condition of a very correct indication. 6--Postoperative, the malignant lesions must be mandatory submitted to the adjuvant treatment. PMID- 12731201 TI - [Acute necrotizing pancreatitis following abdominal trauma]. AB - We present 11 cases of acute necrotizing pancreatitis following abdominal trauma operated in our clinic during the last five years. All patients except one were male. All of them had obvious symptoms of acute diffuse peritonitis when they came to hospital. The diagnosis was established by CT scan in four observations. All operations were performed during 12-36 hours after they came. During surgery, there was complete pancreatic necrosis in seven cases and incomplete in the other four cases. Associated injuries following trauma were represented by: hemothorax- two cases, spleen injury--one case, lesion of mesentery--one case, intestinal lesion--one case. In all cases we performed a wide opening of the lesser sac, pancreatic capsulectomy, necrosectomy and multiple drainage of the pancreatic side (behind the peritoneum) and peritoneal cavity. The postoperative outcome was difficult in all cases. Postoperative morbidity recorded pancreatic leakage (three cases), pseudocysts (two cases) and a perforation of gastric wall. There were 10 cases cured and one patient died. PMID- 12731200 TI - [Present strategies in the diagnosis and treatment of thyroid diseases]. AB - AIM: Selective and critical presentation of the present exploring methods concerning surgical thyroid diseases and accomplishing a pre-surgical diagnosis as accurate as possible. Depending of the diagnosis--choosing the most appropriate surgical strategy according to the newest concepts in thyroid pathology. MATERIALS AND METHOD: Clinical-statistic study, carried out retrospectively over 1,108 patients having various surgical thyroid maladies hospitalized in 1980-1999 period. There have been operated 1,058 cases. Demographic structure as regards sex, F/M was 9.07/0.93 for goiters, whereas for thyroid cancer it was 3/1. RESULTS: Nodular goiters are the most frequent (77.31%), with a pathology benign majority (91.30%). The fundamental problem is the pre-surgical establish malign lesions that require a complex therapeutic protocol comparing to benign nodules. Benign lesions 91.30%, carcinomas 5.48%, others 3.21%. All types of thyroidectomies have been made: T.ST 69.37%, T.T for benign and malign lesions 7.65% and other procedures. Thyroid specific morbidity is 4.25%, whereas unspecific is 8.69%. The 0.08% mortality (1 case), virtually null, place the result between the best of the kind. CONCLUSIONS: Pre-surgical diagnosis, as accurate as possible, based on the clinic exam using the vacuum punction with a fine needle, finally confirmed by the paraffin exam, provided the choice of an optimal surgical strategy, followed by an appropriate post-surgical protocol. Total thyroidectomy is the preferred procedure is most of the thyroid cancer forms and in benign diseases is highly recommended (multi-nodular goiters, hyperthyroid, Basedow disease, cancer). PMID- 12731202 TI - [Duplex Doppler ultrasound value in upper urinary tract obstruction ]. AB - Our prospective study aimed to establish the duplex Doppler ultrasonography (DDU) value in patient's investigative protocol with acute or chronic ureteral obstruction. A. DDU IN ACUTE URETERAL STONE OBSTRUCTION: Between October 1988 and January 2001 we applied DDU evaluation for 177 cases with ureteral lithiasis in which we also studied gray scale echography, resistive index (RI), the difference between affected kidney and contralateral kidney (delta RI) and the characteristics features (amplitude, velocity and frequency) of the ureteral jets (intravesical evaluation in color Doppler mode). a. For 65 cases with RI < 0.70, delta RI < or = 0.06 we didn't find obstruction and 95.4% of the patients had spontaneous passage of the stones. b. For 109 cases with RI > 0.70, delta RI > or = 0.06 the spontaneous passage of the stone was found in 40 cases (36.7%). DDU associated with ureteral jet evaluation increased the predictive value of this method. So, the presence of the ureteral jet on the obstructed side of the upper urinary tract was associated with 63% of spontaneous passage of the stone while the absence of the jet was followed by 5% of spontaneous passage of the stone. The RI was sensitive in 74% and specific in 81% and delta RI was sensitive in 85% and specific in 91%. B. DDU IN CHRONIC URETERAL OBSTRUCTION: Between October 1988 and January 2001 we evaluated the DDU value in chronical ureteral obstruction on 78 cases with operated and/or irradiated genital neoplasm's and retroperitoneal involvement. Our results found RI sensitive in 63% and specific in 70% and delta RI was sensitive in 72% and specific in 78%. After double J stenting (indwelling) RI varied from 0.74 to 0.66 (0.08), with a significative decrease (over 10%) in 61.5% of the cases in which preoperator RI was greater than 0.70. After 1 month we found an increase of RI with 0.02 and after 3 months with 0.05. IN CONCLUSION: We consider DDU a functional imagistic method with an important place in the investigative protocol of the obstructive uropathy. PMID- 12731203 TI - [Controlled stomach fistula for acute operated ulcer. Case report]. AB - The authors analyze the case of a 65 old woman which was hospitalized for sigmoidian stenosant and haemorrhagical neoplasm, confined to the colic wall, without peritoneal or hepatic metastases, and without peritoneal or parietal invasion. Surgical management included sigmoidectomy and termino-terminal anastomosis for reconstructing intestinal transit followed by peritoneal drainage. In early postoperative stage the aspect of generalized peritonitis occurs and there is suspicion of anastomotic fistulae. On surgery, acute and perforated gastric ulcer is found, located in close vicinity to the cardia, on the anterior side of the stomach. Suture of the perforation is undertaken with drainage of the peritoneal cavity, but successfully because fistulization of the sutured perforation followed. Under the given circumstances controlled drainage of the gastric fistulae was carried out, using a Folley probe extended through the fistulae orifice and through the anterior abdominal wall, lateral to the median incision. The blowing of the intragastric balloon and the setting into tension of the gastric wall to the front abdominal wall allowed the sealing of the fistulae route but it took about three months. This technical contrivance has afforded good postoperative evolution and recovery of the patient, who after five years from surgery is in a good condition and has no subjective complaints. PMID- 12731204 TI - [Cystic congenital dilatation of main biliary duct]. AB - A case of cystic congenital dilatation of hepatocholedoch, in a patient aged 27, is presented. The malformation affects the entire biliary duct, being classified as Todani Ic type. The diagnosis was intraoperatively established and the surgical solution was the quasitotal extirpation of the hepatocholedoch followed by the reconstruction of the biliary transit through hepaticojejunostomy on Y loop Roux. The postoperative evolution was favourable and the result is still good after 13 years. PMID- 12731205 TI - [Late results (30 years) after bilateral simultaneous lung resection for chronic bronchiectasis]. AB - The author present a patient operated in 1971 for bilateral bronchiectasies, with bilateral simultaneous pulmonary resections with 11 1/2 pulmonary segments resected, at 13 age. After 30 years the patient is very well with ex. C.T. and pulmonary functions quasi normal. PMID- 12731206 TI - [Dedifferentiated liposarcoma of the omentum]. AB - The authors present the case of a 56 years old patient, from the surgery department, with the diagnostic: "Tumoral abdominal mass". Ultrasound investigation and CT-scan suspicioned a malignant omental tumor. Intraoperator, we found a tumor with macroscopic malignant features (sarcomatous like), greow in omentum; the large border of the stomach being fixed but not invaded by it; the dorsal parietal peritoneum of the right lumba and the little omentum were involved, too. We removed all of the sarcomatous tumor, large omentum, small omentum and the gastric fixed segment. Also, the parietal dorsal peritoneum with metastasis, was surgically removed. The patient's postoperator evolution was very good. The histological report established the diagnostic: "dedifferentiated liposarcoma of the omentum". We considered the case to be interesting because of the difficulties in diagnostic and the rarity of it. PMID- 12731207 TI - [Burch procedure through laparoscopic surgery. Operatory technique]. AB - The authors describe the technique of extraperitoneal laparoscopic Burch colposuspension equivalent to the classical open approach. Patients who undergo laparoscopic Burch procedure experience less postoperative discomfort, shorter hospital stay, quicker recovery and better cosmetic appearance. Based on our initial experience, laparoscopic colposuspension appears to be a viable alternative to abdominal colposuspension. PMID- 12731208 TI - [The parietal reinforcement in the treatment or the prophylaxis of the post operator eventrations]. AB - The authors recommend an original treatment method of "post incision hernias" (eventrations), used in Braila and Sibiu--2 important surgical centers, in more of 100 cases, like retort to classic or modern methods, prothesis or not. The relapses absence and utilization in diverse residence loss grades, as well as the method utilization possibility in the presence of thread granuloma, oblige us to recommend mates studying and critique the used technique. PMID- 12731209 TI - [A new concept in esophageal reconstruction with colon (considerations on 329 operated cases)]. AB - The author reports a personal series of 329 cases of coloesophagoplasty from various etiology, the majority of them carried out for corrosive pharyngoesophageal strictures (276), followed by malignancy (44). A personal philosophy is exposed, based on more flexibility in the choice of the type of colic interposition depending on the pattern of blood supply. This mobile policy called "balanced operation" is opposed to the classic rigid approach based on the use of a single procedure in coloesophagoplasty. His choice is the ileocecum with long ileal loop (65 cm), favoring preservation of the ileocecal valve, and confering an antireflux mechanism. In particular cases the cecum may be removed and an ileal graft carried out. If this arrangement is not feasible one should slide toward the left in a clockwise direction performing a left colon interposition (iso- or antiperistaltic). Overall mortality was of 16 cases (4.9%). Morbidity is analysed and different particular arrangements like continuous colic loop, superlong graft, Roux en Y procedure are described. A general outline of pharyngeal reconstruction in corrosive strictures is presented. PMID- 12731210 TI - Epiaortic ultrasound and intraoperative transesophageal ecocardiography for the thoracic aorta atherosclerosis assessment in patient undergoing CABG. Surgical technique modification to avoid cerebral stroke. AB - BACKGROUND: Cerebral complication during coronary artery bypass surgery (CABG), still occurs in spite of cardiopulmonary bypass (CPB) technology advances. Our study sought to assess the role of epiaortic ultrasound (EAS) and intraoperative transesophagial ecocardiography (TEE) in detection of patients with atherosclerosis of the thoracic aorta and high risk for cerebral embolisation. Consequently our surgical strategies modification for a better cerebral protection. METHODS: Our lot study comprise 352 patients undergoing EAS and TEE, during CABG. EAS was performed before opening of the pericardium using a 7.5 MHz ultrasonic probe packed in a sterile bag. In the presence of moderate aortic atherosclerosis (maximum aortic wall thickness of 3 mm) primarily single aortic cross-clamping and choose of aortic cannulation place was carried out. In cases of severe aortic atheroma (maximum aortic wall thickness > 5 mm or mobile plaque), aortic no touch technique, off-pump were used. From 34 patients with severe ascending aortic atherosclerosis 22 of these patients (18 male, 4 female, age 72(57-79), Parsonet Score 11(0-8), Euroscore 8(2-13), McSPI Stroke Risk Index 6 (1-30%) were managed on "no touch technique", Off-pump. All patients received at least one left internal mammary artery (LIMA) in situ graft and additional extraanatomical bypass conduits: venous Y-graft from IMA (n = 14), arterial Y graft from LIMA (n = 3), vein graft from axillary artery (n = 3), vein graft from the RIMA stump (n = 3). Median operative time was 240 (115-435) min. RESULTS: We found a mild aortic atherosclerosis in 151 patients (43%), moderate in 167 (47%) and severe in 34 patients (8.8%).. Operative mortality was 2/22 in a group with high risk index, from another cause than cerebral stroke. No cerebral stroke occurred. The rate of perioperative myocardial infarction (CKMB > 50U/l) was 5/22. Median stay in ICU, 54 hours (15-1245 hours). After a median follow up period of 8 months one death occurred from cerebral stroke and no myocardial infarction. CONCLUSION: Accurate detection of atheroma on ascending aorta and aortic arch by a combination between EAS and TEE and in special surgical technique modification using off-pump revascularization and extraanatomical bypass for the management of a heavily calcified aorta can result in a very low stroke rate despite a considerable stroke risk. The hospital mortality and morbidity can be lower than predicted by Euroscore and Parsonet Score. PMID- 12731212 TI - Applications of robotics in surgery. AB - The end of the 20th century brought an increased use of computerized technology in medicine and surgery. The development of robotic surgical systems opened new approaches in general and cardiac surgery. Two leading robotic companies, Computer Motion, Inc. and Intuitive Surgical, Inc. have developed the Zeus and Da Vinci respectively, as very effective tools for surgeons to use. Both of them consist of a surgeon console, located far from the operating table, and three robotic arms, which reproduce inside the patient's body the movements performed by the surgeon at the console. The advantages of robotic surgery over laparoscopy and open surgery include: better eye-hand coordination, tremor filtration, steadiness of camera, 3-D vision, motion scale, more degrees of freedom for instruments etc. Of course, there are also some disadvantages, like the lack of tactile feedback, long time of set up, long learning curve, high cost etc. However, the advantages seem to overcome the disadvantages and more and more operations are conducted using robots. The impact of robotics in surgery is therefore very promising and in the future it will probably open even more new ways in the surgical practice and education both in Romania and across the globe. PMID- 12731211 TI - [Serial repeated laparatomy in severe trauma]. AB - Multivisceral trauma and exanguinating hemorrhage lead to hypothermia, coagulopathy and acidosis. Formal resections and reconstructions in these unstable patient is often result in irreversible physiologic insult. For the patients with life-threatening injuries the staged control and repair of injuries may be a saving surgical strategy. The initial phase of "damage control" involves an abbreviated laparotomy, which entails temporary hemorrhage control, perfusion of vital organs and avoidance of enteric or urinary spillage. The surgical procedure is rapidly terminated, with emphasis on a temporary physiologic equilibrium rather than anatomic integrity. That is, the damage control surgery represents an extension of resuscitation phase of trauma in the operating room. The second therapeutic phase involves standard resuscitation and control of hypothermia, coagulopathy and acidosis, combined with surveillance and management of the abdominal compartment syndrome. The last phase involves the definitive repair of all temporized injuries, homeostasis, vascular reconstruction and abdominal wall repair. PMID- 12731213 TI - [The Fitz-Hugh-Curtis syndrome in laparoscopic surgery]. AB - The Fitz-Hugh-Curtis syndrome was diagnosed intraoperatory at 7.1% of the laparoscopic cholecystectomies in our clinic. The attitude in all cases was to perform a complete adhesiolysis. The reasons we consider that support this are: 1. the adhesions that fix the liver to the diaphragma do not allow the surgeon a comfortable access to perform cholecystectomy; 2. if these adhesions are torn accidentally during operation it could end up to the glissonian sheath rupture and uncontrollable bleeding; 3. adhesiolysis might be imposed in order to introduce the ports under visual control; 4. the traction determined by the perihepatitis process against the parietal peritoneum could be responsible for postoperative right quadrant pain; 5. the hepatodiaphragma adhesions make impossible the suction of the intraoperative secretions. A complete adhesiolysis allow a correct lavage of the suprahepatic area preventing the possible retention of clots, bile or even calculi. We didn't notice neither a longer duration of the intervention due to adesiolysis nor intra/or postoperative complications. PMID- 12731214 TI - [Diabetic gangrene]. AB - The large variety of the lesions in diabetic gangrene which require different therapeutical decisions explains our attempt to determine an useful way for the approach of the surgical treatment. Our aim was to avoid both an underestimation of the lesion, which may lead to an insufficient procedure, followed by reinterventions, as well as an overestimation--generating avoidable amputations. We have elaborated a scale of indices of therapeutical prognosis (ITP), using a computer programme, with the main risk factors. The result was a scale between 1 and 10; the value is proportional to the severity of each lesion. We have also analyzed the fiability of ITP on a prospective study including 72 patients with diabetic gangrene. The result was 93%, which has encouraged us to consider ITP useful in establishing a therapeutical attitude. PMID- 12731215 TI - [Cefepime (Maxipime) treatment efficacy in surgical patients ]. AB - Between September and December 2001 in the "Caritas" Surgical Clinic of Bucharest has been conducted a clinical study for the efficiency of Cefepime (Maxipime) treatment in surgical patients. Introduced in therapy in the last decade of the XXth Century, Cefepime (Maxipime) is the most active 4th generation cephalosporin, due to its extended spectrum of activity and its high resistance against beta-lactamases. Cefepime (Maxipime) has a very large spectrum, including the majority of the microorganisms implicated in surgical infections: Enterobacter, Klebsiella pneumoniae/speciae, Proteus mirabilis, Bacillus fragillis, Pseudomonas aeruginosa, Serratia, Citrobacter and other Gram-negative bacilii, Gram-positive cocci (Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus pyogenes). The clinical study included 30 surgical patients, the selection criteria being the severity of the present infection or the potential risk after major (abdominal) surgery. We introduced Cefepime (Maxipime) as first choice of monotheraphy, except: severe, life threatening nosocomial infections, when we associated Cefepime (Maxipime) with aminoglycosides; failure of another antibiotheraphy schema, when we associated Cefepime (Maxipime) with aminoglycosides; suspicion of anaerobe contamination, when we associated Cefepime (Maxipime) with metronidazole. The results of our study support the utilization of Cefepime (Maxipime) as the best choice antibiotic in severe surgical infections, especially in the intensive care and surgical units. Cefepime (Maxipime) can be synergically associated with aminoglycosides and imidazoles (metronidazole). PMID- 12731216 TI - [Primary anastomosis in perforated Crohn disease]. AB - Inflammatory bowel disease (IBD) comprises a group of diseases of intestine characterized by chronic inflammation of the bowel with periods of exacerbation and remission. The evolution of the disease needs, in some therapeutic moments, surgeon's intervention. We described the case of a patient hospitalized through emergency service of Military Hospital of Craiova which presented multiple ileo cecal perforations, histological examination proving clinical supposition of Crohn's disease. The authors achieved, on the other hand, some considerations about etiopathogenical, clinical and therapeutic aspects of this disease. PMID- 12731217 TI - [Difficulties in diagnostic of biliary ileus]. AB - Gallstone ileus is a rare disease which can be life-threatening because of misdiagnosis preoperatively. The authors analyze the clinical observations about two women where, because of associated diseases, delayed in presentation to the hospital, volume depleted electrolyte abnormalities and, not in the last place, because of lack in accuracy of paraclinical investigations, the diagnosis was mad only intraoperatively. CONCLUSIONS: The appearance of clinical setting of high intestinal obstruction in associating with extra-kidney azotemy in female patients with gallstone in their history and no previous abdominal operations must we make to thinking about gallstone ileus. PMID- 12731218 TI - [Cortico-suprarenal carcinoma]. AB - The authors show the case of a 69 years old male with a large corticosuprarenalian tumor that was detected on an random abdominal echographic examination. The patient was operated in the General Surgery Department. of Prof. Agrippa Ionescu Hospital, Bucharest. We performed ablation of the large left suprarenalian gland malign tumor with left nephrectomy, splenectomy and partial pancreatectomy. The hystopathological examination reveals a diffuse corticosuprenalian carcinoma. The case is interesting because of low incidence of this kind of malign tumor and also of the unusual tumor evolution in a long time up to its large size (12 cm in diameter). PMID- 12731219 TI - [The laparoscopic procedures on abdomen with adhesions]. AB - The scar abdomen is more and more seldom a contraindication of the laparoscopic approach. According to Rohr it is classified in scar abdomen after Mac Burney, suprapubic or supraumbilical approach and of the polyoperated patients. Our trial consists in 452 patients with laparoscopic operations consecutive open abdominal surgery (out of the 3900 patients undergoing celioscopic procedures between 1995 2001). The laparoscopic procedure was performed distant to the previous operation in 95.1%, on a neighboring organ to that initially conventionally operated in 4.1% or on the same organ in 3 patients (0.8%). We lead no preoperative fatalities, but the conversational rate was 12% (54 patients), due to the high risk dissection or to the unsolvable bleedings by laparoscopic means (2 cases). The operative accidents consisted in visceral injuries soloed by laparotomy. Postoperatively we registered 5 port site seronas, 1 deep vein thrombosis and 1 pneumonia. The evolution and the mean hospitalization was the same for the scar abdomen patients with the operative procedure accomplished laparoscopically as for non previously operated patients. The data support the feasibility of the laparoscopic procedures on scar abdomen, using the "open laparoscopy" with a reasonably increased conversational rate. PMID- 12731220 TI - [Diagnosis of diaphragmatic hernia]. AB - Diaphragmatic hernias (congenital and traumatic) belongs to thoracoabdominal surgery which is a borderline chapter. Considering frequency, they are on the second place in the diaphragmatic pathology, after hiatal hernias. The author presents the criterias of the clinical examination, based on the bibliographic datas: also by presents the imagistic investigations used for identification of the diaphragmatic hernias, excepting the oesophageal hiatus hernias. There are some particular features appearing in the diagnostical algorithm, too. PMID- 12731221 TI - [Conversion in laparoscopic surgery]. AB - AIM: The conversion causes evaluation in dynamics. METHODOLOGY: The study is a retrospective analysis of the conversion to open surgery in 1993-2001 period, indifferently of the moment and the determinant cause. The yearly dynamics of the conversions was divided by operation types and surgeons. There were also analysed the moment and the cause of the conversion. RESULTS: There were realised 3961 laparoscopic operations (by 7 experienced surgeons and a lot of young surgeons), with 244 conversions (6.2%), percentage variable depending of the operation (3.3% in hernioraphies, 5.3% in cholecystectomies, 8.2% in gynecologic procedures, 12.1% in appendectomies, 33% in abdominal esophagus procedures, 33% in splenectomies) and on surgeon (until 0% and 8%); the differences until the surgeons don't depend on their experience and for the same surgeon, the experience accumulation doesn't reduce the conversion rate. The most conversions happen after a simple inspection or a minimal dissection (73.1% in cholecystectomy) caused by the existence of plastron, the discovery of a difficult anatomic situation or of another pathology; more rarely, the conversion happens in the principal time (23.4% in cholecystectomy), doing to hemorrhage, impossible dissection, visceral injury or even at the end of the operation (3.5% in cholecystectomy), doing to hemorrhage, loss piece or calculs. CONCLUSIONS: The conversion rate depends especially on the correctness of the indication of laparoscopic approach and not on the surgeon experience, what proves that it is a moment of surgical maturity. Decide from the beginning, in the moment of the recognition of a difficult situation and not after the occurrence of a complication, modifies neither the morbidity, nor the much discussed hospital stay. PMID- 12731223 TI - [Small bowel tumors -- peculiarities of diagnosis and evolution]. AB - The authors analyze 13 cases of small bowel tumors. There are emphasized the difficulties of diagnosis, which is rarely established before the laparotomy. In the same time, clinical features are few and operations are often performed too late, in emergency, due to tumors complications (perforation, bleeding, occlusion). There are also presented new etiopathogenic hypothesis, recently mentioned in literature. Surgical treatment is followed by many post-operative complications. Five-years survival is low, under 40% of patients. PMID- 12731222 TI - [Elements of gynecology pathology in girls before and during puberty age]. AB - PURPOSE: The annexial pathology in little girls is representing a more and more frequent form of acute and chronic abdomen, especially in girls around the age of puberty with dysfunction of the menstrual cycle correlated with disturbance of growing and the occurrence of secondary sexual characters. If this kind of pathology is frequent in the period of puberty, there are cases diagnosticated at small ages. MATERIAL & METHODS: The authors are communicating a number of 25 cases with anexial pathology some of them manifesting the symptoms of the acute abdomen (most of them with torsion of the normal or pathological annexes) others having a chronic aspect. The clinical examination correlated with the imagistic investigations were the main elements for the diagnosis. RESULTS & CONCLUSIONS: The results were good, the follow-up of the patients is including social therapy with the girls and their families, for integration in the family, evaluating the chance of giving birth. The occurrence of some forms of acute abdomen was frequent on a pathologic annexes (malformated annexes), causing sometimes the excision of it, extended to the uterus. The presence of a malign tumour can be related with different malformations, also caused by exposure to some risk factors: radiation, pollution, chemical agriculture, deficitary alimentation, stress etc.). PMID- 12731224 TI - [Primary retroperitoneal tumors -- the experience in the Urological Clinic of the Central Military Hospital]. AB - Is a review of the 10 years experience in Urological Clinic of Central Military Hospital concerning the diagnosis and treatment of retroperitoneal tumors, containing personal conclusions about this relatively rare, heterogeneous entity. PMID- 12731225 TI - Effect of beta-lactam antibiotics on the in vitro development of resistance in Pseudomonas aeruginosa. AB - OBJECTIVE: To investigate whether stepwise selection of resistance mutations maz mirror the continued bacterial exposure to antibiotics that occurs in the clinical setting. METHODS: We examined the in vitro development of resistance to a number of commonly used antibiotics (cefepime, cefpirome, ceftazidime, cefataxime, piperacillin and imipenem) in Pseudomonas aeruginosa, a significant nosocomial pathogen. Stepwise resistance was assessed by serial passage of colonies located nearest to the inhibition zone on antibiotic-containing gradient plates. RESULTS: The lowest frequencies of spontaneous resistance mutations were found with cefepime and imipenem; these drugs also resulted in the slowest appearance of resistance of spontaneous resistance mutations. In five wild-type P. aeruginosa strains, cefepime-selected isolates required a mean of 30 passages to reach resistance; resistance occurred more rapidly in strains selected with other cephalosporins. P. aeruginosa strains that produced beta-lactamase or non enzymatic resistance generally developed resistance more rapidly than wild-type strains. For most strains, resistance to all antibiotics except imipenem correlated with increased levels of beta-lactamase activity. Cross-resistance of cephalosporin-selected resistant mutants to other cephalosporins was common,. Cephalosporin-resistant retained susceptibility to imipenem and ciprofloxacin. CONCLUSIONS: From our in vitro study, we can conclude that the rate of development of resistance of P. aeruginosa is lower with cefepime compared with other cephalosporines. PMID- 12731227 TI - [Stapled esophago-jejunal anastomosis after total gastrectomy in gastric neoplasm]. AB - A number of 1031 patients with gastric cancer have been admitted in the First Surgical Clinic of Iasi between 1986-2001. The operability index was 86%, 52% being resectable and 36% having potential curative resection. In this group, 222 patients benefited from total gastrectomies. We consider the introduction of stapled eso-jejunal anastomosis (38 patients) as the most important moment in this development, as the manual performance anastomosis is considered difficult due to the problems imposed by suturing in a deep space. Postoperative course of patients who benefited from stapled anastomosis was simple although many patients had a poor biological status. We encountered only one small fistula, which was dealt with conservative. Patients in this group had immediate postoperative results significantly better than the comparative group with manually performed anastomosis. Introduction of stapler techniques increased the proportions of patients who benefited from palative total gastrectomies for advanced gastric cancer. We consider that a method to solve the frequent complications, insured an increased intraoperative comfort, reduced the number of postoperative complications, increased survival and improved patients quality of life. PMID- 12731226 TI - [Breast tumor in teenager girl. Clinic-pathological considerations]. AB - AIM: Reporting a rare condition in children, with a dramatic clinical appearance preoperatively, though confirming the high frequency of the benign structure of such conditions at this age. MATERIAL AND METHOD: We are reporting the case of a teenager girl, 13 years old, admitted to the Surgical Dept., Children's Hospital in Brasov, for an enlargement of her right breast by a huge mass. Considerations over the diagnostic methods and the surgical management are made, as well as over the pathological aspects and the outcome of the patient. RESULTS: Both clinical evaluation and the imagistic diagnostic-ultrasound scan and mammography-suspect the presence of a large breast mass, but with a rather benign aspect, which is confirmed by the postop pathological exam as a fibroadenoma without any sign of malignancy. CONCLUSIONS: This is to confirm once more the benign character of the majority of the breast masses with this age, despite their volume and appearance, and the opportunity of as much conservative surgical treatment as possible for the pediatric and adolescent patients. PMID- 12731228 TI - [Percutaneous treatment of the liver hydatid cysts under sonographic guidance]. AB - This paper is a retrospective study of the first 51 cases of liver hydatid cysts, which underwent a conservative treatment between April 1996-December 2000. There were 28 females and 23 males with a mean age of 40.1 years (7-65), which had 63 liver hydatid cysts. In the right liver lobe were located 46 cysts, in the left liver lobe were located 7 cysts and in 10 cases cysts were located in both liver lobes. In 4 cases a pulmonary hydatidosis was associated. Abdominal ultrasound and CT scan were routinely performed and the cysts were classified in type I and II after Gharbi's classification. All of these 63 cysts were treated by ultrasound guided fine-needle percutaneous puncture with aspiration and instillation of sterile alcohol 95 degrees. Pre and postoperative the patients were treated with mebendazol or albendazol. They were followed-up by ultrasound and CT scan examination in the second day postoperative and monthly. No new cysts were noted during an average follow-up of 14.7 months and maximal cyst diameter decreased with minimum 72%. Two episodes of reversible anaphylaxis were encountered. The mean hospital stay was 3.3 days. In this paper the indications for conservative treatment and preliminary results of this method are discussed. PMID- 12731229 TI - [Experimental model of orthotopic uterus transplantation in the laboratory rat]. AB - This study develops and standardizes an experimental model of uterus transplantation in the laboratory rat. Twelve orthotropic uterus transplantation were done. Animals were randomized in three groups. Postoperative survival was 100% and 75% at 72 hours. Recipients were euthanased at 24 hours, 48 hours and 72 hours and the grafts were harvested. Patency of the microsurgical anastomoses was 100% at 24 hours, 63% at 48 hours and 0% at 72 hours. The explanted uterine grafts were fixed in formalize and analyzed under light microscopy. The acute allograft rejection starts during the second day after transplantation. In additional dissection, anatomy of the pelvic region with regard to the topography of the uterus, tube and ovarian vessels was studied. This model of uterus transplantation in rats proposes a standardized tool for further research regarding cellular mechanisms of the acute allograft rejection and, for future, pregnancy of the transplanted uterus. PMID- 12731230 TI - [Problems and difficulties in patients with esophageal reconstruction]. AB - The esophageal reconstruction, independent of the manner and indication represents a major challenge for any surgical team. The goal of the present study is the retrospective analysis of cases together with the technical possibilities of surgical approach, viewed through the diagnostical and therapeutical point of view. We analysed retrospectively 154 patients (140 post-caustical stenosis and 14 neoplasms) who have took benefit of reconstructive esophageal surgery, operated in the last 21 years (1981-2001) in the clinic of surgery, of "Saint Mary" Hospital. Although the esophageal substitution has multiple indications (malignancy, benign stenosis, iatrogenic fistulas, reflux disease with peptic stenosis, etc.), in our clinic the technique was performed for neoplasic lesions and post-caustical corrosive stenosis. In the first period of the study (1981 1990) the most utilised method of surgical approach was the gastric tubulisation after the Gavriliu I or II procedures; in a second period (1990-2001) it is obvious that the tendency was to use the colon or the whole stomach as principal visceras for reconstructions. About the immediate results, the majority of post operative complications where minors, the most frequent problem being of pleuro pulmonary nature. We have registered two deceases of patients with corrosive pathology and no post-operative mortality in the first 30 days in neoplasic patients. We evaluated the post-operative functional results using an evaluation score. The anastomotic functionality was good or excellent in most cases (80%), especially in non-malignant reconstructions. For the selection of the most suitable reconstructive procedure, a variety of factors must be explored and evaluated. After a long period in which the use of the colon, in different technical manners, represented the "golden standard" in esophageal reconstruction, we have observed a reevaluation of this attitude and also the more and more frequent utilisation of the whole stomach as esophageal substitute, especially in malignancy. PMID- 12731231 TI - [Cefepime (maxipime), large spectrum 4th generation cephalosporin, resistant to beta-lactamases]. AB - As a result of the appearance of bacterial strains resistant to 3rd generation cephlosporin, since 1993 cephalosporins of 4th generation have been developed and introduced in therapy; among them are: cefepime and cefpirome. Cefepime is the most active 4th generation cephalosporin possessing the following advantages over the 3rd generation cephalosporins: high intrinsic potency due to rapid penetration into the periplasmic space; an extended spectrum of activity that includes many Gram-positive and Gram-negative organisms; Activity against multi resistant Gram-negative bacteria, including Enterobacter and Klebsiella species; Low potential for beta-lactamase induction, especially Bush group 1 beta lactamases, even ar low periplasmic concentrations; Minimal selection of resistant mutant strains. Its spectrum is very large being very active against Gram-negative bacilii: Enterobacter, Pseudomonas aeruginosa, Klebsiella pneumoniae, Serratia, Citrobacter, Proteus mirabilis and less active against Bacillus fragillis. Cefepime is also very active against Gram-positive cocci: Staphylococcus aureus (methicillin-susceptible strains only), Streptococcus pneumoniae, Streptococcus pyogenes. Some of the methicillin-resistant strains of staphylococcus are susceptible to cefepime; Enterococcus is resistant. Due to its high resistance against beta-lactamases Cefepime (Maxipime) is the best choice in life threatening nosocomial infections occuring in patients in the intensive care units. Cefepime can be synergically associated with aminoglycosides and fluoroquinolones. PMID- 12731233 TI - [The difficult laparoscopic cholecystectomies--technical problems]. AB - Referring to the difficult laparoscopic cholecistectomies, the author underlines the actuality of theme and the value of the dialogue between surgeons. The author discusses the anatomo-pathological formes, generating of intraoperative difficulties and emphasises that such historical interventions for difficult cholecistectomies, may be reactualised, with good results in practice. Punctually, he refers to some variants of cholecistectomy: cholecistendesis for sclero-atrophic cholecist and Pribram mucoclasic for cirrhotic patient. The author invites to debate these technical modalities which can evitate or limit, for determined situations, intraoperative risks or failure causes. PMID- 12731232 TI - [Analysis of 3100 laparoscopic cholecystectomies]. AB - AIM: To evaluate the results of laparoscopic cholecystectomy (LC) in the 8 years period. PATIENTS AND METHODS: First LC in Coltea Hospital was performed in September 1993 and introduced for treatment of patients with gallbladder disease. From September 1993 to February 2001 LC was performed in 3100 patients. Mean age 51.2 years (ranged from 8 to 87 years) among 2512 women and 588 men. 232 (7.48%) of the cases were patients with acute cholecystitis. Intraoperative cholangiography was performed in 112 cases (3.6%). RESULTS: Conversion to open cholecystectomy (OC) was necessary in 111 patients (3.58%). Operative complications occurred in 16 (0.5%) patients: CBD lesions in 4 (0.12%) patients, bleeding from cystic artery--12 (0.38%) patients. In one patient CBD injuries was recognized at the time of operation and after conversion to OC primary ductal repair was performed. Postoperative complications occurred in 44 (1.41%) patients: a) local infection--in 15 (0.48%) patients (subhepatic abcess-3, wound infection-9. b) bile leakage--in 21 (0.67%) patients. c) haemoperitoneum because of the bleeding: from the abdominal wall at the trocar insertion site--in 2 patients, from a. cystica-one patient. d) obstructive jaundice due to stone in CBD--in 5 patients (endoscopic papillosphincterotomy and stone extraction was performed). There 21 reoperations due to complications: 13 laparatomies and 8 relaparascopies. Two patients (52 and 64 years old) died after LC-mortality 0.06 per cent. Mean hospitalisation day was 3.8. CONCLUSIONS: To prevent iatrogenic CBD injuries correct preparation with a clear identification of the anatomic structures is essential. Relaparascopy and endoscopic retrograde cholangyopancreatography can be successfully used in the treatment of complications after LC. PMID- 12731234 TI - [Laparoscopy in abdominal trauma. Preliminary report in 37 cases]. AB - The role of diagnostic laparoscopy (DL) and therapeutic laparoscopy (TL) in abdominal trauma is not clear. Even after diagnostic punction lavage (DPL), ultrasonography (US), and CT scan (CT), in some cases is difficult to decide between laparotomy and observation. In 37 cases of abdominal trauma, a laparoscopic evaluation was done; 28 abdominal blunt trauma (22 associated with multiple trauma), and 9 abdominal wounds (8 stab wounds). In blunt abdominal trauma, DL was done for haemoperitoneum (26 cases), after DPL, US or CT. In three cases, with equivocal diagnosis, an extra abdominal operation in general anaesthesia was necessary. In abdominal wounds a DL was done for suspicion of penetration. All the patients were haemodynamic stable, TS > or = 12. A laparotomy was necessary in 12 cases (32.43%), a LT was possible in 5 cases (13.51%). In the rest of cases, a DL with or without lavage-drenage enough. PMID- 12731235 TI - [Surgical procedures in penis cancer]. AB - In the period of 1980-1999 (20 years) in Colentina Surgical Clinic were admitted and operated 16 patients with the diagnosis of pennial cancer (histopathology- squamocellular carcinoma), 13 of whom being from Colentina Dermatological Clinic. The mean age was about 64 years old, 10 cases presenting phimosis. Surgical procedures undergone were represented by partial pennial amputation with bilateral superficial inguinal lymphadenectomy (10 cases), total pennial amputation with superficial inguinal lymphadenectomy (4 cases), respectively emasculation with bilateral inguinal-iliopelvic lymphadenectomy (2 cases). In the postoperative period 3 patients presented lymphedema and lymphorrhagia, solved by conservative management. All the patients benefited of favorable evolution on the period they have responded to the controlled postoperative follow up (1-3 years). PMID- 12731236 TI - [The diagnostic strategy in abdominal trauma]. AB - Diagnosis of a deep visceral lesion in an adult following abdominal contusion is almost always difficult as the abdominal signs are later or masked by associated lesions. All of the controlled studies agree on the superiority of peritoneal puncture-lavage over metrosonography or CT scan in the diagnosis of homeoperitoneu. The diagnostic performance of CT, TDM in the identification of lesions solid organs is very much better that that of ultrasonography. The diagnosis of an isolated lesion of a hallow viscus benefited from progress in imaging to a much lesser degree an peritoneal puncture lavage with leucocyte count remains the key examination. The diagnostic strategy must be based on the patient's clinical condition and on this hierarchy of complementary investigations. PMID- 12731237 TI - [The primitive gastric non- Hodgkin lymphoma]. AB - THE PURPOSE: Of this work is to point out diagnostic problems together with the surgical indications in primitive gastric non-Hodgkin Lymphoma (LGNH). THE MATERIAL AND METHODS: Consist of 11 cases of LGNH (3.09% of 350 gastric cancers that underwent surgery between 1991-2000) of which 4 were women and 5 men, of an average 53 years of age. Three of the cases underwent surgery for various complications (HDS, perforation and pyloric stenosis) or for other clinical forms such as the pseudo-ulcerative one (4 cases) and the gastric carcinoma mimicking form (4 cases). Preoperative diagnosis was established by means of endobiopsy in 5 cases. SURGICAL TREATMENT: total gastrectomy (4 cases), inferior polar subtotal gastrectomy (5 cases) and 2/3 gastric resection (2 cases). RESULTS: Complications involving sub-phrenic abscess that triggered re-intervention in 2 cases; no immediately postoperative mortality. THE DISCUSSIONS AND CONCLUSIONS: Are enumerating the difficulties we encountered in diagnosing the cases and especially in the preoperative histologic diagnosis, the staging of the disease and setting the treatment with an emphasis on the surgical treatment. THE COMPLICATIONS: Induced by the disease, the diagnosis uncertainty and the early stages of the disease are as many eligible indications for the first linje surgical treatment in LGNH. PMID- 12731238 TI - [Recurrent dermatofibrosarcoma protuberans]. AB - The present paper tends to emphases by one hand the necessity of the histological examination, as a routine exam, on every surgical procedure, even minor ones, and by the other hand, the difficulties of the differential diagnostic, including by histological means, in lesser importance lesions. In this line we will present the case of female patient who had a lesion labeled as a postoperative granuloma and it proved to be a malignant neoplasia. PMID- 12731239 TI - [Intestinal tuberculosis--cause of acute surgical abdomen]. AB - Tuberculosis, in its various forms, remains an important cause of morbidity and mortality in developing countries in immunodeficitary patients. The indicatives of epidemiology of tuberculosis show that Romania presents a fresh outbreak of the disease in the last few years. The purpose of this paper is to present from the various forms of extrapulmonary tuberculosis, the intestinal tipe which have a high incidence. The authors describe theirs preliminary experience of intraoperative small and large bowel emergencies resections in a short period (1 year) of three young patients (between 30 and 40 years old) with history of pulmonary tuberculosis. The pathology was complex (bowel obstructions, peritonitis) and so were the surgical operations (resections, devirations). The patients showed short and long term good results. PMID- 12731241 TI - [The 10 principles of the camera person (or what a surgeon must do so that another one can see what he does)]. AB - Miniinvasive videoendoscopic surgical techniques represent a revolution in all surgical fields and generate deep changes. Many principles of open surgery were put in doubt, some of them been considerate like dogma, the operating setup was changed, some mentalities and beliefs were modified and a new member of the surgical team appeared. This specialty doesn't exist in open surgery and its importance could not be minimized. This new kind of surgeon has particular knowledge and its actions during the laparoscopic procedures are different in opposition to open procedures. The following article refers at laparoscopic procedures but we are sure that exists several common points with others miniivasive techniques and some of this ideas are generally valid. PMID- 12731240 TI - [Intestinal obstruction caused by abdominal pseudotumoral endometriosis mimicking a peritoneal carcinomatosis--case report]. AB - A 40-year-old woman was admitted in emergency condition with the symptomatology of bowel obstruction. Intraoperative findings consists of a pelvic fixed tumoral mass, and numerous other tumors spread in the whole abdominal cavity mimicking a peritoneal carcinomatosis without liver metastases. Three of the tumors where about 4 cm in diameter producing stenosis of the terminal ileurn and sigma. We considered the case as it was a peritoneal carcinomatosis caused by an uterus or ovarian cancer and we decided for palliative surgery, performing ileo transversostomy and sigmoidostomy above the obstruction. The histopathologic findings from more pieces of tumors revealed endometriosis without cancerous changes. Postoperative the patient underwent cytostatic and then hormonal therapy. After 3 month the CT scan revealed an important reducing in volume of the pelvic tumoral mass and the barium enema didn't showed any stenosis under the colostomy so, we closed it extraperitonealy. At 16 month after the first operation the patient was reoperated for a parietal defect. At the second look we found no tumors. The pelvic tumoral mass has disappeared, the uterus seemed to be normal but two big ovary cysts were present. We performed bilateral adnexectomy and the repair of the parietal defect. Postoperative evolution was favorable without any complication or complains at 3 month after the last operation. PMID- 12731242 TI - [Duodenum preserving cephalic pancreatectomy and pancreatico-gastrostomy in the surgical treatment of the chronic pancreatitis]. AB - Cephalic pancreaticoduodenectomy had been introduced in surgery practice by O. Whipple, for the treatment for the cancer of ampulla of Vater, later this indication has also been extended to other pancreatic disorders including the cases of chronic pancreatitis. Cephalic pancreatectomy with the remaining of the duodenum used lately in the treatment of chronic pancreatic eliminates the disadvantages of the operation Whipple. Further on, we present a case of chronic pancreatitis where a cephalo-pancreatic resection has been done with the remaining of the duodenum, the pancreatic blunt being anastomosed with the stomach by pancreatico-gastrostomy, terminal-lateral, a cystoduodenostomy had being done to this patient. Ten months after the surgery, the patient is in a good general state, without subjective pain, putting weight, and a glucose metabolism without changes. CONCLUSIONS: Pancreatico-gastrostomy may represent a modality of solving the pancreatic blunt after the cephalic pancreatectomy with the remaining of the duodenum. PMID- 12731243 TI - [Surgical treatment of thyroid nodules. The immediate results after different thyroidectomy methods]. AB - The extension of the resection for thyroid nodules depends both on nodules' nature and immediate or late postoperative complications risks. This clinical study analyzed the immediate complications appeared after partial thyroidectomy comparatively with those developed after total thyroidectomy. We studied 1411 patients operated in two clinics (from Romania and from France) which have two different attitudes concerning the width of the resection. Paralysis of recurrent laryngeal nerve occurred in 1.0% of patients with partial thyroidectomy and 3.0% of patients with total thyroidectomy, while only one patient (0.6%) developed permanent hypoparathyroidism after total thyroidectomy. In conclusion, total thyroidectomy can be performed by experimented surgeons with a recurrent or parathyroid injury risk similar to partial thyroidectomy. However, the surgeon should take into account the patient survey capacity and the discomfort produced by life substitutive treatment. PMID- 12731244 TI - [Enterostomy post emergency enterectomy]. AB - This is a clinical approach regarding 43 resection of intestine, performed in emergency condition, terminated as enterostomy, with represent 20% of enterectomyes performed in emergency condition and 1.6% of urgent operations. The decision of enterostomy has been taken in the conditions of peritoneal sepsis, occlusion or the association of the two circumstances. The results are comparatively analyzed between the cases with enterostomy that has been made from the beginning (66% success, 33% gone wrong), and those with enterostomy made at the second intervention (14% success, 86% gone wrong). One discusses problems of leading, technique and post-operating nursing. The intestinal reintegration has been made possible at 16 patients after a timing of three of four weeks. PMID- 12731245 TI - [Acute intra-abdominal hypertension and "abdominal compartment syndrome"]. AB - The acute intra-abdominal hypertension causes profound physiologic abnormalities, both within and outside the abdomen. Just as in compartment syndrome in the extremities, gut mucosal ischemia begins long before clinical signs are evident, explaining the name of "abdominal compartment syndrome" given to the acute, markedly increased intra-abdominal pressure. The abdominal compartment syndrome was initially described in patients with severe abdominal injuries and massive transfusions and crystalloid infusions, caused by the closure of fascia or skin under tension, the use of bulky abdominal packs to control diffuse bleeding, the massive bowel distension and edema, and the continued bleeding into the abdominal cavity. Intra-abdominal pressure can be monitored by measuring the urinary bladder pressure with a manometer, connected to the transurethral Foley catheter, with the symphysis pubis as the zero point. A persistent elevation of the intra abdominal pressure beyond 20-25 cmH2O, with significant respiratory, hemodynamic and renal dysfunction is an indication for abdominal decompression, before the manifestations of abdominal compartment syndrome became clinically evident. The mortality in patients with abdominal compartment syndrome is over 40%, even when adequately treated. PMID- 12731246 TI - [Hepatic serial resection. Portal vein resection followed by hepatic resection in second time]. AB - Hepatic resection still remains the only potential curative treatment for either primary or secondary malignant liver tumors. In order to increase the resectability of initially considered non-resectable tumors and to decrease the posthepatectomy morbidity and mortality, ligature of a portal branch with consecutive hepatic resection is recommended. The ligature of a portal venous branch was performed in 12 patients with gross hepatic tumor: hepatocellular carcinoma (2), peripheral cholangiocarcinoma (5), hepatic metastases from colorectal cancer (5). Two-stage hepatectomy was performed in 5 patients. The interval between the two operations ranged between 4 weeks and 6 months. Hepatic resection could not be performed in 7 cases due to the loco-regional progression of the disease (4 cases) or to the absence of the hypertrophy-atrophy process (3 cases). Hepatic failure occurred posthepatectomy in 2 patients, resulting in the death of one of the patients. Two patients died at 5 and 10 months respectively while two other patients are still alive, free of recurrence at 6 and 12 months respectively. In conclusion, portal vein ligature can be considered in selected cases of unresectable gross hepatic tumors that can be eventually, resected in a second operation. The two-staged hepatectomy is not always feasible. Moreover, the hypertrophy of the controlateral lobe does not always prevent the postoperative hepatic failure.. PMID- 12731247 TI - [Post-bulbar ulcer, features of diagnosis, tactics and surgical conduct]. AB - From the diversity of places of duodenal ulcers, those postbulbars represent particular anatomo-clinical forms, due to the implication of pancreas and liver, accompanying by chronic pancreatitis, relation with papilla, multitude of ulcers and their serious complications (giant ulcers, hemorrhages, penetrations). Our experience is based on 324 patients with PBU which have been operated during 30 years (1970-2000), which represent 7.4% from the total number of duodenal ulcers operated in the same period. Men/women rate is 8/1. Ages between 40-60 years old plead in favor of vascular element (hypoxia) in the determination of lesion of PBU. The medical treatment being usually reflector definitive the absolute surgical indication which needs special tactical and technical approaches, taking into consideration the particularities of duodenum, implication of pancreas, gallbladder and papilla. In the surgical treatment of PBU we used most frequently the operation Pean-Billroth I--128 cases (39%), followed by the conservative operations in association with vagotomy--122 cases (34.5%). We have got good postoperative results in using of the exclusion operations--42 cases (13%) and exclusion resections--Finstere-Plenk--26 cases (8%). Preservation of duodenum in the digestive transit was possible in 89% of cases. General postoperative mortality was 1.5%, in hemorrhages the postoperative mortality was 4.1%. PMID- 12731248 TI - [Surgical initial attitude in epithelial ovarian advanced cancer]. AB - The purpose of this work is to show the value of aggressive excisional surgery in epithelial ovarian advanced cancer (COEA). MATERIAL AND METHOD: Between May 1993 May 1997, there were 103 obs. with ovarian diseases; 28 obs. (27.1%) with COEA and 75 obs. (72.9%) with benign diseases. All the patients had a subtotal hysterectomy with bilateral anexectomy and omentectomy. The operation was extended to other anatomic elements; in the case of 4 obs., we practiced a segmentary resection of left colon, 2 obs. had an anterior resection of the rectum and a partial resection of the pelvic peritoneum, 15 obs. suffered an excision of tumoral masses from the gastro colic omentum and the right parietocolic space and 7 obs. had a resection of pelvic peritoneum and of the right parietocolic space peritoneum. The optimal cytoreduction with a tumoral residuum less than 2 cm was realized for 23 obs. (82.14%) and for 5 obs., the tumoral residuum was greater than 2 cm (17.76%). The medium survival of the patients with a tumoral residuum inferior to 2 cm was 41 months, at 4 years 54% of the observations being alive, compared to only 14 months for the patients with tumoral residuum superior to 2 cm, at 4 years the survival being only of 15%. All the observations have beneficiated of chemotherapy. The disease reappeared for 23 obs. after a variable period of time. It's treatment consisted of: surgery followed by chemotherapy for 18 obs. and chemotherapy of second line alone for 5 obs. CONCLUSIONS: The ovarian epithelial cancers, famous for an intermediary answer to chemotherapy, lead to an aggressive surgical management with partial extra genital organ sacrifices, to offer a chance of life. PMID- 12731249 TI - [Volvulus of the wandering spleen]. AB - The volvulus of the floating spleen is a rarity generally manifestated by an acute painful abdominal syndrome located into the left flank. The preoperative diagnosis can be eased off by an abdominal ecography which shows the spleens volume and its absence from the spleenlodge. Further on, we present the case of a female patient (28 years) who during hospitalization has developed a spleen volvulus being preoperative diagnosticated and terminated by splenectomy. Referring to Abell only in 4% of all cases you can diagnosticate this preoperatively. Recent studies also mention the conservative, laparoscopical treatment which means a laparoscopical detorsion and normal dyeing of the spleen followed by a splenopexy with a tent. PMID- 12731250 TI - [Mini-invasive treatment, through endoscopic papillo-sphincterectomy, of post surgery biliary fistulae]. AB - External biliary fistulas associated both with open and laparoscopic biliary surgery, need a quick solution to prevent local and general complications. Open surgical treatment was the only solution until the minimally invasive techniques, such as endoscopy, were developed. The authors present 10 cases of postoperative external biliary fistulas, successfully treated by endoscopic papillosphincterotomy alone, technique which is compatible with other endoscopic procedures of treatment, performed in association with endoscopic papillosphincterotomy or not. PMID- 12731251 TI - [Combined laparoscopic and endoscopic treatment of gallbladder and bile duct stones]. AB - In the present study we have tried to find what is the best time for endoscopy in the treatment of gallstones associated with common bile duct stones. METHOD: We have selected on the intention to treat 89 patients suspected of cholecysto choledocholithiasis. There have been 38 cases with preoperative endoscopy (Group A), 35 cases with postoperative endoscopy (Group B) and 16 cases with perioperative endoscopy (Group C). RESULTS: In group C it has been a significant higher proportion of successfully treated cases (94%) and a lower hospital stay (8.6 +/- 3.7 days). CONCLUSIONS: Combining the endoscopy and laparoscopy in the same operation ("rendez-vous" technique) is the best approach for treating cholecysto-choledocholithiasis. PMID- 12731252 TI - [Is there a learning curve for laparoscopic cholecystectomy ?]. AB - The use of laparoscopic surgery it is growing rapidly in Romania. We have tried to find if a learning curve for laparoscopic cholecystectomy exists and we have evaluated our training program to insure the quality of health care. METHODS: There where analyzed 2585 procedures performed by 22 surgeons. The relationship between operative incidents/accidents and laparoscopic experience was evaluated. With a regression model we have find the tendency of these relationship. RESULTS: Surgeons appear to learn this procedure rapidly. CONCLUSIONS: Training curses with hands-on experience with animal models and proctoring from an experienced laparoscopist at the first 30 operations is needed, for each individual surgeon, to insure the quality of health care. PMID- 12731253 TI - [Observations regarding lymphatic gastric drainage in malignant gastric pathology through intra-operatory lymphography]. AB - The authors became interested in the lymphatic drainage of the stomach in order to attempt to determine for each region the type of drainage and the risk of cancer spread. They studied 50 dissection, including the stomach, 115 cases records of gastric carcinoma, 8 lymphographies by ultrafluid lipiodal, and 48 preoperative injections of vital colorant. The system of drainage proposed by Rouviere was on the whole confirmed; however, one should emphasize the doubling of the hepatic chain, the existence of long collectors which bypass a relay in the left gastric artery, the importance of the posterior gastric artery which transmits the lymphatic of the splenic chain. Finally, the authors emphasize the existence of 3 longitudinal areas on the stomach where the presence or absence of valvules in the subserous collectors orients the lymph towards the lesser or greater curvature of the stomach, which easily explains the onset of isolated carcinomatous adenopathy, situated on the curvature opposite the neoplasm. PMID- 12731254 TI - [Synchronous colonic cancers]. AB - The aim of this study is to present the difficulty of an accurate preoperative diagnostic for synchronous colonic cancers and to sustain the necessity of total colectomy. A retrospective study was carried out on 16 patients hospitalized in the IIIrd Surgical Unit, St. Spiridon Hospital, U.M.Ph. "Gr.T.Popa" Iasi between 1990-1999. The surgical procedures were: extensive colectomy with ileo-sigmoid anastomosis, segmentary colectomy, total colectomy with ileo-rectal anastomosis. RESULTS: Perioperative mortality: zero; uneventfully recovery for all patients. A metachronous lesion 3 years after the first operation was detected in one patient; postoperatively, one patient developed occlussion 3 months after, requiring re-operation. CONCLUSIONS: 1. synchronous colonic cancers are closely related with a genetic instability of the colonic mucosa; 2. total colectomy is a safe manner to prevent metachronous lesions. PMID- 12731255 TI - [The therapeutical strategy for locally advanced breast cancer]. AB - As for the other cancers, the strategy of therapy of breast cancers is going to a unitary standardization. In our department between 1984-1999 we are operated 1040 patients with breast cancers, which means 25.3% of all cancers treated. 688 (64.3%) were CMLA, 646 (96.7%) of them were in patients women and 22 (3.3%) men. The mean age was 52.4 years (3-84 years). All patients were divided into two trials and analyzed: retrospectively (A) 312 (46.7%) and prospectively (B) 356 (53.3%) patients, 51.2% of patients was in III and IV TNM stage. The patients from trial B were treated concerning with specific therapeutically protocol, adapted by age, anatomopathological form, volume of tumor, skin or thoracic wall invasion, inflammatory lesions, lymph node invasion and physiological period. The results were: the increase of number of radical surgical interventions, the decrease of the morbidity, the increase of survival and a better quality of life. PMID- 12731256 TI - [Errors and difficulties in the diagnosis and management of acute and chronic abdomen in children]. AB - The authors are reviewing on a lot of 2844 cases between 1996 and 2000 the difficult problems of differential diagnosis between acute surgical abdomen in children and intestinal tuberculosis, abdominal tumors and inflammatory diseases such as acute osteomielitis. They are presenting 13 particular cases in which the acute abdomen diagnosis was difficult or even omitted. PMID- 12731257 TI - [Microbiology of nosocomial infections in a general surgery department]. AB - This paper aim is to outline the importance of nosocomial infections, characterized by great incidence, great mortality rate and specific bacteriology, in a surgery clinic. The study include 566 patients that developed 665 nosocomial infections (10.65% incidence), among the 5950 patients that underwent surgical operations in 5 years (1992-1996); 54 patients developed two or three nosocomial infections, which explain the incidence of nosocomial infection greater than number of patients. From bacteriological point of view predominance of Gram negative bacilli (especially E. coli) and pathogen staphylococcus characterized the nosocomial infections. The infection source was the patient himself, previously colonized with hospital specific microorganisms. Every clinical form of nosocomial infections was characterized by the present of specific pathogen microorganisms; the knowledge of these pathogen agents is very important for the antibiotic treatment applied before the bacteriological exam. PMID- 12731258 TI - [Cyst duplication of the colon in an adult. Case report]. AB - The cystic type of the enteric duplication has the same histological structure as the digestive tract and the same mucosal lining as the segment to which is attached. These are rare pathological conditions especially in an adult as they usually are diagnosed and treated in childhood. A 39 year-old women has been elective admitted in our unit complaining of upper abdominal pain and a palpable abdominal mass in epigastrium and left upper quadrant. The imagistic findings (abdominal ultrasound and CT scan) raise the suspicion of a pancreatic cystic lesion. Intraoperative we found a large cystic lesion localized in the transverse mezocolon close to the transverse colon. The resected specimen was examined histopatologically and found to be a cystic duplication of the colon. The patient made an uneventful postoperative recovery. PMID- 12731259 TI - [Medio-thoracic esophageal stenosis, produced by pseudotumoral ulcer]. AB - A non-specific esophagitis developed in a young male, apparently determined by heavy abuse of alcohol. Shortly after, a stenotic pseudo-tumoral mediothoracic esophageal lesion was diagnosed. Although there was a high index of suspicion regarding the caustic origin of the lesion, cancer could not be excluded using current diagnostic methods. Temporary feeding jejunostomy was followed by uneventful total esophagectomy and gastric pull-up. Pathologic examination of the resected specimen was a surprise, revealing an esophageal ulcer based on massive severe non-specific esophagitis. Retrospectively, the ulcer was judged as primitive or idiopathic. We discuss some etiopathogenic aspects of common secondary esophageal ulcers (related or not to reflux disease), in contrast with idiopathic esophageal ulcers. PMID- 12731260 TI - [Small intestine adenocarcinoma. Cause of mechanic obstruction through slide of intestine into the small intestine (clinical case)]. AB - It's present the case of a patient from nineteen years with acute abdominal pain with debut in the day of presentation. Biological investigations is negative, with exception VSH = 40 mm/h. The gynecological examination raise the doubt of a strangled and floating ovary balloon. It's operate in emergency--it's about a mechanic obstacle from the small intestine (adenocarcinoma) with slide the small intestine into the small intestine with three cylinders. Reduction the small intestine slide-segmental enterectomy. Recovery. The mechanic obstacle from the small intestine with slide the small intestine into the small intestine through a cancer, to young patients, is a rare disease. The small intestine adenocarcinoma is meeting particularly at grown-up and old patients (3-4% from the digestive cancers) (2). The case in face is alone in personal statistics. PMID- 12731262 TI - [Serous cysts of the spleen treated by laparoscopic surgery]. AB - The authors present two case of a 25 years and 32 years old male patients with serous cysts of spleen that were detected on an random abdominal ultrasonography. Abdominal pain in the left hypocondrium was only symptom of this patients. The patients was operated by laparoscopic method, with Ultra-Shears and intraoperative ultrasonography examination. This kind of techniques give us the opportunity to performed the ablation of the cysts with preservation of the spleen with fast full recovery. PMID- 12731261 TI - [The reposition technique for treatment of obstruction or migration of the peritoneal dialysis catheter]. AB - Continuous ambulatory peritoneal dialysis are widely used in the management of patients with chronic renal failure. The permanent presence of the catheter into the peritoneal cavity generate a series of specific complications. Two of the most important causes of dysfunctional peritoneal dialysis catheter are obstruction and malposition. Failure to restore the drainage function of the catheter by conservative method should be followed by a surgical procedure: laparoscopic reposition or replacing the catheter. This paper present an original technique which has some major advantages: required local anesthesia; doesn't replace the existing catheter; the dyalysis program could be started very quick after procedure; it is a feasible and reproducible technique. PMID- 12731263 TI - [Surgical treatment of locally advanced esophago-gastric cancer; preliminary results]. AB - The aim of this paper is to sustain the palliative resection in neoplasm of the esophago-gastric junction, as a surgical approach that allows a better post operative life comfort in comparison with simple gastrostomy. 62 observations with proximal neoplasm of the stomach (12.5%) were identified between January 1996-August 2001, representing 12.5% of the 496 patients with gastric neoplasm admitted in our unit in the same period. Out of these 62 cases, 55 (88.71%) underwent surgical procedures. Our attitude was aggressive in 25 cases. 40.32%, including the locally advanced lesions with palliative surgical indications (18 obs.). The other 30 patients underwent: 10 laparotomies, 5 gastrostomies and 15 jejunostomies. Local invasion to the neighboring organs imposed partial resection of the transverse colon--1 obs., of the transverse mesocolon--2 obs., and corporeo-caudal pancreatectomies--3 obs. The surgical approach was a left abdomino-thoracic incision, with total gastrectomy and distal esophagectomy, with N1 and N2 lymphadenectomy, splenectomy, and esojejunal intrathoracic anastomosis, with a Roux-en-Y loop, with or without jejunostomy (13 obs.). The immediate post operative complications were 8 anastomotic leakage, one duodenal stump fistula, one occlusion due to a jejunostoma, and 13 extradigestive complications. There were 5 post-operative deaths. CONCLUSION: Neoplasm of the esophago-gastric junction is lately diagnosed, but whenever is possible, total gastrectomy with distal esophagectomy should be carried out. PMID- 12731265 TI - [Allogenic blood transfusion and postoperative outcome -5-year experience in a digestive surgery unit]. AB - The paper presents a retrospective study of a digestive surgery unit, evaluating the outcome of the patients who were operated and received blood transfusions over a 5 years time, compared with a random sample of patients operated during the same period and who did not receive transfusions. The patients were stratified by the type of operation and the ASA criteria. The paper also briefly reviews the literature on this topic. The study noted a prolonged postoperative course and overall hospitalization time, and an increase in the infectious complications and mortality rates in the study group, compared to the control group. The results of the study support a transfusion practice policy based on strict indications, and the use of alternatives to transfusion, whenever available. PMID- 12731264 TI - [The capacity of preoperative ultrasonography in predicting technical challenges in laparoscopic cholecystectomy]. AB - Ultrasonography is a nonsophisticated, cheap and safe exploration, that makes it the most used non-invasive method in the diagnosis of biliary diseases. Present study aims to evaluate preoperative ultrasonography's capacity of predicting technical challanges in laparoscopic cholecystectomy (LC). MATERIAL AND METHOD: The trial is represented by a sequence of 100 patients undergoing LC in 2001. The conversions to open procedure were excluded. Patients with certified lithaisis were re-examined by ultrasonography the day before surgery. The sonographic features as size, volume (scleroatrophic gallbladder), function (distension, contraction), wall thickness, hydrops, number and size of stones, infundibular position (impactation) of the stones, perivesicular liquid collections hepatic and pancreatic aspects, main bile duct caliber were registered. During LC, the difficulty in performing the procedure was measured using a 10 points-score of following parameters: 1 peritoneal and perivesicular adhesions; 2. Difficult grasping of the gallbladder; 3. Cystic duct's dissection; 4. Cystic artery; 5. Liver bed; 6. Difficult cystic stappling; 7. Gallbladder's wall efraction; 8. Need for intraoperative cholangiography; 9. operative bleeding; 10. Operative time. RESULTS: Gallbladder's and the number, the size and infundibular impactation of the stones was significantly associated to ultraoperative difficulties (p < 0.05), the other parameters having not a sensitive influence upon LC procedure. CONCLUSION: Preoperative ultrasonography is able to furnish valuable data in predicting LC challenges. PMID- 12731266 TI - [Liposuction and lipolysis in breast cancer]. PMID- 12731267 TI - [Intestinal obstruction by biliary ileus; clinical experience and literature review]. AB - BACKGROUND: Gallstone ileus is an unusual and peculiar complication of biliary lithiasis. Less than 1% of gallstones migrate into the gut, causing 25% of non strangulated small bowell obstructions in elderly population. Diagnosis is difficult, leading to late operation. Considering the median age of the patients and the fact that in most cases surgery is delayed, there is a lot of dispute regarding the best approach. Recent technical facilities in diagnostic and surgical practice seem to be irrelevant for the general outcome. MATERIAL AND METHOD: All 6 patients operated since 1981 (median age 67 years, 5 female) were retrospectively analyzed, to determine the most valid option. Biliary ileus was recorded in 0.16% of all operations for billiard lithiasis and 1.52% of all enteric occlusions. There was a mean delay of 4.2 days for onset of symptoms to admission and further 5 days to surgery. Particular elements suggesting the ethiology were absent in most cases. In all cases the gallstone migrated in the duodenum. The obstruction was jejunal (3 patients), proximal ileal (2 patients) and distal ileal (1 patient). Enterolithotomy was practiced in 3 cases, with subsequent cholecystectomy and fistula closure in 1 case; one-stage repair in 2 cases and enterolithotomy with temporary external drainage of the fistula (cholecystostomy) in 1 patient. RESULTS AND CONCLUSIONS: There were no specific (fistular) complications; 1 aged patient with distal occlusion died because of severe metabolic disorders. The most "favourable" cases were those with very large gallstones occluding the jejunum. Smaller stones were less symptomatic, resulting in delayed diagnosis and surgery; the more aboral the occlusion, more indication for abstention regarding primary repair of the fistula. PMID- 12731268 TI - [Therapeutic attitude in acute necrotizing pancreatitis]. AB - The necrosectomy, celiostomy and pancreatic drainage represent the surgical treatment of choice in necrotizing pancreatitis. We present the clinical observation of a patient 59 years old operated in surgical service of Baia Mare for acute necrotizing pancreatitis, discussing the moment of operation, tips of operations, postoperative complications as well as our experience in acute grave pancreatitis treatment. PMID- 12731270 TI - [Ascites--a complication of chronic alcoholic pancreatitis]. AB - We present the case a 44 year old patient previously diagnosed with chronic alcoholic pancreatitis with pancreatic ascites during hospitalization in the Gastro-Enterology department. As the conservative therapy performed for 21 days was not effective in diminishing the ascites, the patient was admitted in our Surgical Department and scheduled for surgical intervention. He was operated and we discovered a small dimension cyst (7/4 cm) developed in the body and tail of the pancreas, fistulized in the peritoneal cavity through an outlet positioned below the insertion of the mesocolonum transversum, fairly close to the duodeno jejunal angle. We executed a cysto-jejunal anastomosis by using the first loop of the jejunum, secured with a politer drainage positioned as in WITZEL technique and drive out in the left upper quadrant. The postoperative evolution of the patient was difficult, but constantly positive. The patient left the hospital 32 days after the intervention. The clinical and ultrasound follow-up after three months were normal. PMID- 12731269 TI - [Thyroid cancer and chronic granulocytic leukemia in the same patient: Cernobyl or coincidence?]. AB - We present a case of thyroidian cancer and chronic granulocytic leukemia simultaneously found in a 28 years old patient. Initial surgical management consisted in total thyroidectomy and removal of the lymph nodes; local recurrence after 6 months required excision of the tumoral block by radical neck dissection. Hematological disease was controlled with Hydroxiureea, associated, when needed, with Busulphan and Cytosar. The patient is in acceptable general condition and free of any local recurrence or apparent metastases 2.5 years after the diagnosis. PMID- 12731271 TI - [Laparoscopic management of peritoneal dialysis catheters]. AB - Continuous ambulatory peritoneal dialysis catheter could be placed also by open laparotomy as well as by laparoscopic techniques. We did a retrospective study on cases to compare the results of laparoscopies. There were included 42 patients which we divided in two groups of 21. Group A underwent 21 cases in which catheters was inserted by open laparotomy. Continuous ambulatory peritoneal dialysis was started in 24 to 48 hours later. Group B incharged 21 patients underwent laparoscopic placement of the catheter between 2000 and 2001. Continuous peritoneal dialysis was started early (after 6 hours). The mean operative time was 28 minutes in group A and 30 minutes in group B. Fluid leakage was noticed in 4 patients in group A and in 3 patients in group B. Peritoneal reactions occurred in 5 patients in group A and in 2 patients in group B. Tip migration occurred in 5 patients in group A (one of which was mobilized accidentaly early after intervention) in which was necessary 4 open reinterventions, and no patients in group B. In group B one patient underwent a simultaneous liver biopsy for cirosis and another female patient underwent ovariectomy for a giant ovary cyst. Laparoscopic placement of dialysis catheter leads to better function than does open procedure, it allows immediate start of dialysis and permits simultaneous performance of other laparoscopic procedures. PMID- 12731272 TI - [Gastric stromal tumor treated by laparoscopic surgery]. AB - The authors describe a 59 years old female patient, with a gastrointestinal stromal tumor located on the posterior wall of the gastric funds, who was treated successfully by laparoscopic wedge resection (with clear resection margins), through an anteriorly placed gastrotomy, thus allowing an endoscopic linear cutter Endo GIA, to excise the tumor with a cuff of normal gastric tissue. The anterior gastrotomy was performed with Ultra-Shears. Delivery of the tumor through the gastrotomy is essential for success. The operative time was 110 minutes. The tumor was diagnosed as a gastrointestinal stromal submucosal tumor (of low-grade malignancy) and immunohistochemicaly, this tumor was positive for CD 34. CONCLUSION: Posterior gastric tumor can be removed using laparoscopic surgery. PMID- 12731274 TI - [Dynamic observation of carbofuran on symbolic enzymes in testis of rats]. AB - To examine the effects of carbofuran on the testis of male rats. The activities of beta-glucuronidase (beta-G), glucose-6-phosphate dehydrogenase(G-6-PD) and lactate dehydrogenase isoenzyme-x (LDHx) in serum and testis homogenate were determined for the rats given carbofuran at the dises of 0.3, 1.5 and 3.0 mg/kg orally for 7, 35 and 77 days. The results showed that after 7 days, the activities of beta-G in serum in all exposed groups were lower than those in control (P < 0.05). The activities of beta-G in testis homogenate in 0.3 mg/kg and 3.0 mg/kg were higher or lower than those in control (P < 0.05), respectively. After 77 days, the activities of G-6-PD in serum both in 1.5 mg/kg and 3.0 mg/kg were lower than those in control (P < 0.05). The activities of LDHx in testis homogenate in 3.0 mg/kg were lower than those in control (P < 0.05). It suggested that exposure to carbofuran could testis damage. PMID- 12731273 TI - [Laparoscopic suture or open suture in perforated duodenal ulcer]. AB - Between Nov. 1994-Jan. 2001 we performed laparoscopic suture with omentoplasty of perforated duodenal ulcer (PDU) in 51 patients out of 56 it was intended (this constituted the laparoscopic group--LG). The selection criteria were young patients, age < 40 years, no associated diseases, onset of the operation under 12 hours from the occurrence of the perforation, absence of clear ulcer history. In the same period, we performed an open suture based on the same criteria in 105 patients (open group--OP). The results showed a difference between needed analgesia (2.8 days for LG vs. 5.2 days for OG) and a hospital stay of 6.1 days in LG vs. 7.7 days in OG. The incidence of postoperative complications was 5.88% in the LG and 7.61% in the OG with 1.96% and respective 1.90% reoperation rate. In conclusion laparoscopic suture of PDU with associated postoperative modern therapy of ulcer disease could be the treatment of choice in young patients. PMID- 12731275 TI - [Estrogenic activity of some environmental chemicals]. AB - The study was designed to explore the estrogenic activity of environmental disrupters [n-4-noniphenol (NP), bisphenol A (bisA) and dibutylphthalate (DBP)]. Estrogen-respective breast cancer cell line T47D was grown in DMEM medium containing 10% bovine serum. Five days before the addition of the test compounds, the cell were washed in phosphate-buffered saline, and the medium was substituted with a phenol red-free DMEM medium containing 5% dextran charcoal-stripped FBS. The respective test compound was added in fresh medium and the control cell received only the vehicle (ethanol). The proliferation of T47D cells was analyzed by MTT assay, 3H-TdR incorporation assay and flow sytometer. The results indicated that, compared with the EtOH control cell, the test compounds (n-4 noniphenol, Bisphenol A and dibutylphthalate), like estradiol, markedly enhanced the proliferation of T47D cell and the metaphase of cell division, and the results showed time-dependent and dose-dependent model. These data showed that the tested chemicals could enhance the proliferation of human cervix cancer cell in vitro. This might hinted that these chemicals possessed estrogenic activity and they might play their estrogen through estrogenic receptor. PMID- 12731276 TI - [Study of vitamin A nutritional status and the correlation of vitamin A and iron in school-age children]. AB - To understand the VA status, the detectable rate of the sub-clinical VA deficiency and the correlation of VA and iron in the rural school-age children of Beijing Mountain Area. In the dietary survey, blood samples were collected by venopunction and serum VA, and hematological index were determined in 305 children 7-13 years. The intakes of dietary energy, protein and iron were > 85% of the RNI and AI, but dietary intake of VA were (513.7 +/- 286.1) microgram to be 59.7% of the RNI; The average concentration of serum VA was (1.01 +/- 0.29) mumol/L, 59.0% of subjects have serum VA lower than 1.05 mumol/L, in which, 12.8% of them were lower than 0.70 mumol/L, and only 41.0% of subjects were normal levels of serum. With aggravation of iron deficiency from ID to IDA phase, the average serum VA concentrations were descending from 1.02 mumol/L in ID phase to 0.97 mumol/L in IDA phase. But the trend has no statistical significance. The results showed that sub-clinical VA deficiency was 59.0% in the rural school-age children, so VA deficiency was a main nutritional problem in rural school-age children of Beijing mountain area. PMID- 12731277 TI - [Zinc deficiency on pathological changes of femur epiphyseal growth plate in rats]. AB - To study zinc deficiency on pathological changes of femur epiphyseal growth plate in rats thirty Wistar rats were randomly divided into three groups: zinc deficient group (Zd), control group (Ctr), and pair-fed group (Zp) to study the effects of zinc deficiency on femur epiphyseal growth plate of rats and the mechanism involved. After 8 weeks feeding, the histomorphology of right femur indicated that chondrocytes in the epiphyseal growth plate of Zd group were generally ill-organized and mis-shaped. The number of chondrocytes decreased. Trabecular bone in the epiphyseal of Zd group were also ill-organized, scarce and slim. The cavities of marrow in Zd animals were significantly larger than those of Ctr and Zp. The volume of trabecular bone, the mean trabecular plate density of Zd rats were significantly decreased, but their mean trabecular plate space was significantly increased. In addition, the Zd animals had significantly lower concentration of osteocalcin and growth hormone in sera. The above results suggested that zinc deficiency impaired proliferation and differentiation of chondrocytes and balance between osteoblast and osteoclast function by reduce growth hormone levels in sera. The lack of zinc in diet resulted in disorder of bone molding and ultrastructure. PMID- 12731278 TI - [Study on the oxidative injury of ECV304 cell induced by homocysteine]. AB - The mechanism of homocysteine-induced oxidative injury on ECV304 cell line, a spontaneously transformed immortal human endothelial cell line, was studied. ECV304 cells were incubated with different levels of Hcy (0.05-2.00 mmol/L) for 12 hours. The activity of major anti-oxidative enzymes and the generation of reactive oxidative species (ROS) were measured by chromatometry and fluorochromy. The results showed that Hcy could induce obvious oxidative injury on ECV304 even at low level (0.05-0.10 mmol/L). The level of MDA was increased and the active oxygen was generated in the cells. The activities of anti-oxidative enzymes (SOD and GSH-Px) except the activity of CAT were decreased in the cells. It was concluded that Hcy may induce oxidative injury on EVC304 cell line through increasing the levels of ROS and inhibiting the activities of anti-oxidative enzymes. PMID- 12731279 TI - [Effect of Zingiber OfficinaleRosc on lipid peroxidation in hyperlipidemia rats]. AB - Zingiber Officinale Rosc(ginger) is the food of rhizoma species as well as Chinese traditional medicine and has various pharmacological effects. The last researches showed that ginger not only reduced plasma lipid levels but also the mouse atherosclerotic lesion areas. The ginger antioxidative effect maybe pay an important role in attenuation of development of atherosclerosis. Antioxidative effect of Zingiber Officinale Rosc on hyperlipidemia rats have been studied and the changes of GSH-Px and LPO in their blood have been observed in this paper. Male adult Wistar rats were grouped into control, preventive and curative teams. The experimental teams were respectively fed on the test diet containing 2% ginger and 5% ginger, in order to measure the changes of plasma lipid peroxides (LPO) and glutathione (GSH-Px) after the experiment. The results show that ginger increased GSH-Px and reduced LPO in the rats' blood. Ginger could inhibit and/or scaving radicals of rat body in different degrees. PMID- 12731280 TI - [Study on the immune response of big-ear white rabbit and Luoman hen to ochratoxin A]. AB - Antibodies of anti-ochratoxin A (anti-OTA) were prepared by immunizing white rabbits and Luoman hens. The immune responses to OTA of these two animals were compared. It was concluded that the immune response of white rabbits was faster than Luoman hens'. The anti-OTA IgG of rabbits was produced after 30 d while the anti-OTA IgY was produced after 60 d. The top titre of anti-OTA IgG was 3-4 times of the titre of anti-OTA IgY's. PMID- 12731281 TI - [Detection of bovine-derived materials in import animal feeds and food by PCR assay]. AB - A PCR assay for detection and identification of bovine--derived materials in import animal feeds and food is developed. A 271 bp sequence from bovine specific mitochondrial DNA is amplified. The amplified specific sequence of the bovine mtDNA from meat and feed samples is demonstrated by both direct sequencing and restriction endonuclease digestion yield analysis. This method can detect bovine mtDNA in those less than 0.125% of bovine meat. Owing that this method is specific, rapid and sensitive, it can be utilized as a routine control assay in prevention of spreading of bovine spongiform encephalopathy in China caused by imported food and animal feeds especially meat and bone meals. PMID- 12731282 TI - [Method on PCR detection of bovine materials in animal feedstuffs]. AB - The reliable, simple and easy to operate methods were established to extract DNA from animal feedstuffs. A set of PCR primers were designed for bovine specific mitochondrial DNA sequence. With this method, PCR can be successfully utilized to evaluate the presence of bovine materials in animal feedstuffs. A set of commonly used primers of 18Sr-RNA were designed as endogenuous reference gene, and check on the quality of template extraction from animal feedstuffs, and avoid the result of false negative. PCR amplification condition was one cycle of 96 degrees C for 2 min, then 35 cycle of (94 degrees C for 40s, 60 degrees C for 50s, 72 degrees C for 60s), and finally one cycle of 72 degrees C for 5 min. PCR amplified specific gene sequence 271 bp for bovine materials from animal feedstuffs. PMID- 12731283 TI - [Study on the degradation of residual insecticides on and in the vegetables and washing off method]. AB - The degradation of residual insecticide in the vegetables is according to the first order of chemical dynamitic reaction. The half-life of insecticide on/in the vegetable can be calculated through a series sampling and analysis the residual amount to obtain a regression equation or by a simplified equation. Although the total amount of residual insecticide degraded according to the first order of chemical dynamitic reaction, the lost of the partial insecticide which absorbed in the leaves seems likely according to a more complicated exponent curve which has been described. The adsorbed residual insecticide (on the leaves) was easy washed away. The non-ionic detergent is more effect than water. But the absorbed insecticide (in the plant tissue) is hard to wash out. PMID- 12731284 TI - [Study on changing regularity of leptin, estradiol and testosterone during peripuberty in girls]. AB - To study the changing regularity of leptin during puberty in girls and the relationship of leptin, estradiol and testosterone each other, serum levels of leptin, estradiol(E2) and testosterone(T) were determined in blood in 150 obese girls, 150 normal girls and 150 malnourished girls, aged from 7 to 17 years old. Their ages of menarche were surveyed. The results showed that serum levels of leptin increased with age, and had double peaks. The first peak of leptin was found during the early phase of puberty in all groups. The value of peak were 14.46 micrograms/ml, 8.85 micrograms/ml, 8.60 micrograms/ml in obese, normal and malnourished groups respectively. The second peak of leptin emerged at the end of puberty in the three groups. The level of E2 increased sharply at the eleven years old in three groups, then increased progressively with age. Serum level of E2 did not correlated with leptin. There was a peak level of T at 10 years old in obese groups, as well as normal and malnourished groups. The level of T emerged rise sharply at 12 years old after a decrease in all groups. Serum level of T did not correlated with leptin. It is suggested that leptin may be a more important decisive factor during pubertal development course in girls. The first peak of leptin may be the signal of onset of puberty. The second peak of leptin may be the signal of the end of puberty. PMID- 12731285 TI - [Effect of different vitamin E homologous analogues on human hepatoma cell HepG2 proliferation in vitro]. AB - To examine the effect of alpha-tocopherol(alpha-T), gamma-tocopherol(gamma-T), delta-tocopherol(delta-T), and vitamin E succinate (VES) on the proliferation of human hepatoma cell (HepG2), cell proliferation was detected by hemocytometer count, MTT assay and flow cytometry (FCM) with different VE homologues analogues (alpha-T, gamma-T, delta-T and VES) and different concentrations (12.5 mg/L, 25 mg/L, 50 mg/L, 100 mg/L and 200 mg/L). The results showed that The growth of HepG2 cells was inhibited by delta-T and VES at the dosages of 12.5-200 mg/L in comparison with the negative control group, while gamma-T showed weak effect of inhibition and alpha-T did not show any inhibition effect. The dose range to produce inhibition effects varied with different analogues. A dose-dependent inhibition of cell growth was found in HepG2 cell lines treated with different vitamin E homologues analogues. An accumulation of cells in G0/G1-phase and a significant decreasing of cells in S-phase were found as evaluated by flow cytometric analysis employing a PI-staining method. It is suggested that delta tocopherol and VES decreased HepG2 cells growth and viability. A dose-dependent of anti-proliferation was found in HepG2 cells line. the order of efficiency of four vitamin E analogues was delta-tocopherol > VES > gamma-tocopherol > alpha tocopherol. The proliferation was blocked in S-phase. The difference in nature and magnitude of the anticancer effects does not correlate with their reported relative antioxidant activity and might be due to minor differences in their structure important to their biological activities. These results suggest that delta-tocopherol and VES could be promising anti-hepatoma agents. PMID- 12731286 TI - [Regulation of a new amino acids preparation on the expression of growth factors in traumatized rats]. AB - The effect of a new amino acids formulas on wound healing and the expression of several growth factors in traumatized rats were investigated. Twenty-four male Wistar rats were randomly divided into control group, 17AA group and 20AA group after operation, and fed with rations containing tyrosine, a commercial amino acids (17AA), and a new amino acids formula(20AA) respectively. Body weight, dietary intake, the hydroyproline and tensile strength of wound were measured. At the same time, the expression of EGF, TGF beta 1 and bFGF in small intestine were also determined. The results showed that the levels of hydroyproline in sponge and tensile-strength of wound were higher in 20AA group than in 17AA group(P < 0.05). The orders of expression of EGF, TGF beta 1 and bFGF in small intestine were as follows: 20AA group > 17AA group > control group. It is concluded that the new amino acids preparations can enhance the expression of growth factors in small intestines and promote wound healing. PMID- 12731287 TI - [Prevalence of hypertension and the relationship between dietary pattern and hypertension in rural middle-aged and elderly in HeNan Province]. AB - 6186 subjects (age > or = 40) from a cross sectional study named Dietary, Life Style and Chronic Diseases cooperated by Chinese Academy of Preventive Medicine and BioSignia Inc. in 1997 were included in this analysis. The prevalence rate of hypertension and the dietary intakes were analyzed. The results showed that the prevalence rate was 44.8% in the corresponding, which were obviously increased compared with the results in 1991, and were higher than other areas in this same period in China. The dietary pattern was obviously unreasonable in this population. The prevalence rate of hypertension was higher in the population who eat less vegetable, fruit, fishes or eat more salted vegetable. And the results indicated that improving dietary pattern may be an inexpensive and effective way to prevent hypertension and to promote people's health in rural area. PMID- 12731288 TI - [Effect of calcium supplement on superoxide dismutase and malonaldehyde of disuse osteoporosis in young rats]. AB - Effects of Ca supplement on the bone mineral density(BMD), red blood cell superoxide dismutase(SOD), plasma malonaldehyde(MDA) of disused osteoporosis were studied in this paper. Twenty five healthy pregnant SD rats were randomly divided into 5 groups, groups 2-5 were reproduced with patterns of animal osteoporosis. The diet in groups 1-2 was the basic diet, while the groups 3-5 experiment diets were supplemented with Ca. Selected 3 young rats, live with the mother rate each brood. The BMD, red blood cell SOD, plasma MDA were measured 37 d later. The results showed that BMD level of young rats in Ca supplement groups is higher than vigorous groups (P < 0.05). The MDA in vigorous group was higher than Ca supplement, groups(P < 0.01) and SOD was lower than high biologic calcium group(P < 0.05). It was suggested that Ca supplemented certain effect on red blood cell SOD, plasma MDA of disused osteoporosis. PMID- 12731289 TI - [Determination of chlorhexidine acetate in disinfectant by high performance liquid chromatography]. AB - A high performance liquid chromatography(HPLC) was described for the determination of chlorhexidine acetate in disinfectant on a C18 column using acetonitrile-0.02 mol/L potassium dihydrogen phosphate solution(pH2.5) (40:60) as mobile phase. The regression equation was linear when the chlorhexidine acetate concentration was lower than 1.00 g/L. The detection limit was 8 mg/L. The precision was less than 4.0% and the recovery varied from 99.8% to 115.2%. The method was more sensitive, rapid and simple than other HPLC methods, and the characteristic of low disturbance makes it especially important for the assay of chlorhexidine acetate in compound disinfectant comparing with titrimetry and UV spectrophotometry. PMID- 12731290 TI - [Study on the regulative effect of isomaltooligosaccharides on human intestinal flora]. AB - OBJECTIVE: In order to study the effect of Isomaltooligosaccharides on Human and mouse intestinal flora in vivo. METHODS: 1. 30 health subjects (15 male, 15 female) were randomly selected from the clinic of Tianjin CDC. They were provide 15 g Isomaltooligosaccharides once a day for 7 days; 2. 40 BABL/c mice were divided into 4 groups, 3 experimental groups and 1 control group. These 3 experimental groups were fed different dose of Isomaltooligosaccharides by gavage for 7 days. Feces of both human and mice were determined before and at the end of the experiment. RESULTS: The results showed that the reproduction of Bifidobacteria and Lactobacillus greatly increased, and the growth of Clostridium perfriengenes was significantly inhibited both in mice and human intestinal tracts. CONCLUSION: Isomaltooligosaccharides could regulate and improve the intestinal flora in both mice and human intestine. PMID- 12731291 TI - [Lactobacillus casei microbiological assay of plasma and RBC concentrations of folic acid with 96-well microtiiter plates]. AB - To investigate the plasma and RBC concentrations of folic acid in normal persons. Microbiological assay was used to determine the plasma and RBC folic acid in 59 normal persons by using of 96-well microtiter plasma. The results showed that the plasma and RBC folio acid concentration were (8.2 +/- 2.9) microgram/L and (337.5 +/- 91.0) microgram/L in male and female normal persons (aged 18 to 32) respectively. The coefficient Variations determined within assay and between assay were 2.2% and 4.7% respectively. This method may reduce reagent costs, shortened the assay time, increase the sensitivity and improve the reproducibility. It can be used for large scale survey. PMID- 12731292 TI - [Role of apolipoprotein A-IV in the regulation of food intake]. AB - Although many exciting functions of Apolipoprotein A-IV(apo-A IV) have been proposed, this review focuses on its unique role in regulating food intake. Apo-A IV is a glycoprotein produced by human intestine and rodent intestine and liver, but the small intestine is the major organ responsible for the circulating apo-A IV. Our Laboratory recently demonstrated that apo-A IV protein and apo-A IV mRNA were present in rat hypothalamus. Intestinal apo-A IV synthesis is markedly stimulated by fat absorption and the formation of chylomicrons. Intestinal apo-A IV synthesis is also enhanced by the peptide tyrosine-tyrosine. The inhibition of food intake by apo-A IV is mediated centrally. Apo-A IV likely plays a role in the short-term regulation of food intake. Other evidence suggests that apo-A IV may also be involved in the long-term regulation of food intake and body weight. Reduction of apo-A IV in response to lipid feeding in both animal and humans that chronically consume a high-fat diet may explain in part why the chronic ingestion of a high-fat diet predisposes to obesity. PMID- 12731294 TI - [A new toxicological concept-acute reference dose]. AB - Acute reference dose (ARfD) is a new concept in toxicology and risk assessment, and mainly used for assessing health effect from short-time exposure to environmental chemicals. The ARfD of a chemical was defined as "an estimate of a substance in food or drinking water, expressed on a body weight basis, that can be ingested over a short period of time, usually during one meal or one day, without appreciable health risk to the consumer on the basis of all the known facts at the time of the evaluation. It is usually expressed in milligrams per kilogram of body weight." ARfD is different from ADI and reference dose. In this review the principles and methods in establishing ARfD were explained. PMID- 12731293 TI - [Biomolecules associate with iron homeostasis]. AB - There are many biomolecules that take part in the regulation of iron homeostasis. Most of these literatures were published in the magazines of molecular biology. These molecules were studied individually. In this review, these molecules were viewed as a whole in the maintenance of iron balance. It classified these molecules into those which are associated with iron absorption, those which are associated with iron transport, which store excessive iron and which monitored the iron homeostasis. PMID- 12731295 TI - [Flow injection analysis technique and its application in food analysis]. AB - The review is based on the achievement and progress reported of food analysis with the flow injection analysis technique during 1990-2000 which contains the fundamental principle, unsolved problem, research and application on FIA technique. PMID- 12731297 TI - [Studies on the zinc bioavailability using stable isotope techniques]. AB - Zinc is one of essential trace elements in humans. Zinc homeostasis is mainly regulated by the changes of intestinal absorption efficiency and endogenous excretion, namely the regulation of bioavailability. Studies on zinc bioavailability play an important role on a thorough understanding of zinc metabolism and reliable assessment on population zinc status. Stable isotopes are valuable tools for research on mineral bioavailability and metabolism. Zinc stable isotopes can be used for all population as tracers with no exposure to radiation. Since 1980s' many different stable isotope methods have been applied to the zinc metabolism research. However, limitations to these approaches and methodological problems remain to be resolved. In addition, the introduction and development of new analytical instruments not only make it possible to use zinc stable isotope, also greatly diminish the difficulty and cost on it. The paper is mainly focused on the zinc bioavailability and present a review of its development history, main research methods and analytical skills. PMID- 12731296 TI - [Review of dietary risk factors for osteoporosis]. AB - Dietary factors are convenient but correctable factors in the pathogenesis of osteoporosis. The peak bone mass in the young can be increased and the rate of bone loss in the elderly possibly be reduced by dietary manipulation, which would be important and beneficial in the prevention of osteoporosis. Dietary risk factors for osteoporosis include low calcium intake, low or high protein intake, low vitamin intake, smoking, and high intake of alcohol, coffee, carbonated beverage and salt. PMID- 12731298 TI - [Franco-Cambodian medical cooperation: mission accomplished?]. PMID- 12731299 TI - [Internal medicine and self-image. Models of beauty or health indicators?]. PMID- 12731300 TI - [Papua New Guinea]. AB - Papua New Guinea is an independent country located in Oceania with a population of 4.9 million. Urban development is low and the estimated population density is 9 inhabitants per square kilometre. The terrain is mountainous and accessibility for health care services is difficult in some locations. Medical care facilities are organized in 18 provinces that are divided into 2 to 8 districts in which basic health care and hospital services are delivered through 1765 first aid units, 319 dispensaries, 189 specialized clinics and centers and 19 hospitals. There are no local schools for training medical and paramedical personnel who come from outside the country. Malaria, filariasis, leprosy, tuberculosis, infant diarrheal diseases, viral hepatitis, and envenomation are major public health problems. Buruli ulcer, dengue fever and arboviruses are emerging diseases. PMID- 12731301 TI - [Histoplasmosis]. PMID- 12731302 TI - [Sodium dichloroisocyanurate: a major disinfectant for drinking water]. PMID- 12731303 TI - [Televised education: experience in Senegal]. AB - As a possible response to the shortage of qualified teachers in Africa, a promising pilot study using television is being carried out at the A. Le Dantec University Hospital in Dakar, Senegal. Trainees at the Dakar Medical School receive long-distance coaching mainly in video-assisted surgery from experts at several partner centers in Europe, namely: Rangueil Hospital in Toulouse, France, IRCAD in Strasbourg, France and Saint Peter's Hospital in Brussels, Belgium. Results support more widespread use of televised courses. PMID- 12731304 TI - [Ramsay Hunt syndrome]. PMID- 12731305 TI - [Directly... from Florence. Third European Conference on Travel Medicine]. PMID- 12731306 TI - [Severe sickle cell disease and pregnancy. Systematic prophylactic transfusions in 16 cases]. AB - Sickle cell disease is a major cause of maternal and neonatal mortality in tropical regions. Treatment consists of regular blood transfusion during pregnancy and delivery. Transfusion may be performed selectively (in function of the clinical status of the mother, fetus, and pregnancy) or prophylactically. Both management strategies result in a significant improvement in the obstetric outcome. However systematic prophylactic transfusion has been shunned due to the risk of transmitting infections and allo-immunization. The16-case series reported here demonstrates a real benefit for prophylactic blood transfusion during pregnancy complicated by sickle cell disease in tropical zones. Improvements in our transfusion curves resulting from better donor selection, follow-up, and phenotyping support arguments previously made over one decade ago in favor of the prophylactic approach especially in countries with high transfusional risks. In our experience the risks associated with more transfusions was offset by the greater safety margin and better efficacy of the prophylactic approach. PMID- 12731307 TI - [Suspected epidemic at a hospital in Senegal]. AB - Most outbreaks of noscomial infection are detected by means of molecular biological testing. However Euclidian distance is a rapid, reliable alternative if facilities for molecular biological testing are unavailable, as at the Principal Hospital in Dakar, Senegal. Over the 7 month period from 01/10/98 to 31/04/99, 110 of the 1033 blood cultures specimens collected in a group of 2360 children hospitalized in pediatric departments A and B of the Principal Hospital were found to be positive for Burkholderia cepacia. This high incidence suggested the possibility of an epidemic. This likelihood was further increased by evidence showing that all strains exhibited the same biotype and by results of pulsed field electrophoresis (CHEF Mapper, Spel digestion) indicating that bacteria was of clonal origin. These findings were entirely consistent with those obtained by Euclidian distance but excessive mortality was not observed in the children involved. Testing of environmental specimens demonstrated the presence of the same strains in respirators and catheters. The most feasible hypothesis to account for these findings is environmental contamination resulting in mucocutaneous contamination of patients. Hemoculture containers were probably contaminated during handling. PMID- 12731308 TI - [Laryngo-tracheo-bronchial foreign bodies in children at the University Hospital Center of Ouagadougou (analysis of 96 cases)]. AB - Over a 10-year period, a series 96 patients were treated for laryngotracheal and bronchial foreign bodies in the ENT department of the Ouagadougou University Hospital Center (Burkina Faso). The purpose of this study was to analyze the epidemioclinical features and therapeutic pitfalls encountered in that series and to propose measures to improve management. The age of patients ranged from 10 months to 14 years. The male-female ratio was 1.7. Anamnesis documented foreign body aspiration was obtained in 77% of cases. The foreign body was organic in 78 cases (81.2%) including vegetal material in 56 cases. Management was delayed due to late diagnosis since only 59.3% of the patients were admitted to the hospital within the 48 first hours. Endoscopic removal was performed in all cases but such procedures remain challenging in developing countries due to a lack of proper anesthetic facilities, skilled personnel and adequate equipment. Tracheostomy was performed in 10.4% of cases mainly in patients with foreign bodies located in larynx. One patient died during extraction. In addition to providing more information and education on prevention of laryngotracheal and bronchial foreign bodies, better management requires better training of medical personnel and improvement of technical facilities. PMID- 12731309 TI - [Uterine rupture of pregnant uterus: study of a continuous series of 63 cases at the referral maternity of Niamey (Niger)]. AB - Uterine rupture contributes significantly to high maternal mortality rates in developing countries. We conducted a prospective study of 63 cases of uterine rupture during pregnancy in the Gazobi Hospital in Niamey, Niger between November 1977 to December 1998. The incidence in the maternity department was 2.3%. Most of the patients were transported from inland areas. Mean age was 29 years, and mean parity was 5.7. More than half of patients (53%) presented uterine scar defects. Two thirds of patients had undergone no prenatal care. Analysis of iatrogenic etiologic factors demonstrated strong correlation with use of oxytocin, forceps delivery and fundal pressure. In 75% of cases, diagnosis of uterine rupture was made before delivery. The main symptom was hemorrhage. Surgical treatment consisted of hysterorraphy in 79% of the cases. Maternal mortality was 4.7% and perinatal mortality was 76%. Discussion focuses on the frequency of uterine rupture in developing countries, etiologic factors, diagnostic modalities, and therapeutic guidelines. Several simple preventive measures are proposed to reduce the incidence of uterine rupture in developing countries. PMID- 12731310 TI - [Uterine rupture in Senegal. Epidemiology and quality of management]. AB - The purpose of this prospective longitudinal study was to analyze data concerning patients treated for uterine rupture at surgical maternity hospitals in Senegal between January 1 to December 31, 1996. A total of 50 cases of uterine ruptures were recorded during the study period, i.e., 1 rupture for every 45 cesarian sections. Typical epidemiological features were rural residence (68%), age over 30 years (66%), multiparity (64%), and presence of obstetrical risks factors (76%). In 96% of cases, rupture usually occurred after failure or natural delivery assisted by personnel with limited skills in non-surgical facilities. Treatment consisted of uterine suture in 22% of cases and obstetrical hysterectomy in 78%. As a result of poor facilities for emergency transportation (68% of cases), mean delay between the indication for operative treatment and intervention was 11 hours. Maternal mortality and morbidity were 16% and 14% respectively. Neonatal mortality was 95%. The incidence of uterine rupture could be lowered by improving emergency obstetrical care and identifying risk factors for dystocia during prenatal examinations. PMID- 12731311 TI - [Tuberculosis and HIV in Bangui, Central African Republic: strong prevalence and management difficulties]. AB - The increasing incidence of tuberculosis in relation with the HIV-AIDS epidemic poses a major public health problem in sub-Saharan Africa. The purpose of this retrospective study was to analyze the prevalence of HIV-1 infection, clinical presentation, and bacteriological findings in patients treated for tuberculosis in a hospital department in Bangui, Central African Republic between January 1996 and December 1998. Among the 1142 patients who benefited for HIV serology, HIV-1 prevalence of was 82% (IC95%: 79-85%). Most patients (92%), had not undergone HIV serology before hospitalization. Mean age was 34 years. Sex ratio F/M was 1.21. Diagnosis of tuberculosis was based mainly on clinical and radiological data. Positive sputum smears were available for only 52% of the patients. The most frequent site of tuberculosis was the lungs with no significant difference between the HIV-positive and HIV-negative groups (83% versus 79% respectively). Sputum-smear examination was positive in 42% of the patients with no significant difference between the HIV-positive and HIV-negative groups (43% versus 37%). Upon admission patients usually presented advanced disease, with 11% dying within a week after hospitalization. There was a steep increase in the prevalence of HIV in tuberculosis-infected patients in Banqui, from 32% in 1988 to 62% in 1994. In spite of the existence of a National Tuberculosis Control Program, diagnostic facilities remain limited and diagnosis of tuberculosis and HIV-infection is often delayed. Outpatient care must be improved. PMID- 12731312 TI - [Study of HIV seroprevalence in patients with pulmonary tuberculosis in 1999 in Chad]. AB - Tuberculosis is the most common opportunistic disease occurring in the course of human immunodeficiency virus (HIV) infection. With the development of HIV infection in Chad, tuberculosis has quickly become a major public health concern. The purpose of this cross-sectional study in two tuberculosis centres (Moundou and Ndjamena) was to evaluate HIV seroprevalence, epidemiological characteristics and risk factors in patients with tuberculosis. All patients with positive sputum smears who had never been treated for tuberculosis previously were eligible. A total of 466 patients (sex ratio M/F: 1.96) were included during the six-month period between January and June 99. Each subject was asked to fill out an anonymous standardized questionnaire with detailed information on demographic characteristics, sexual behavior and other risk factors for HIV infection. Data were compared using the Chi-square test, Student's T test and multivariate analysis (logistic regression). One third (33.2%) of patients was seropositive for HIV-1. Mean age was 31.1 years in HIV-positive group and 33.6 years in the HIV-negative group (p = 0.02). The age groups with the highest risk for HIV were 20-29 years and 30-39 years (p < 0.01). Women were more often seropositive that men (39.5% versus 30.1%; p = 0.04). Seropositive patients were more likely to have multiple sexual partners (mean: 1.76; p < 0.01) and a history of sexually transmitted disease (19.9% versus 8.1%; p = 0.01). The mean age at the time of first sexual relations was 16.3 years in the HIV group and 17.1 years in the control (p < 0.01). The percentage of seropositive patients was 39.6% at Moundou and 29.8% at N'djamena (p > 0.01). Multivariate analysis showed that early age of first sex relation (OR = 0.85; 95% IC: 0.74-0.97), higher number of sexual partners (OR = 1.8; 95% CI: 1.4-2.4) and level of education were strongly correlated with HIV infection. The prevalence of HIV in tuberculosis patients is a good indicator of HIV-infection in developing countries. Prevalence of HIV infection is high in tuberculosis patients in Chad. Surveillance for tuberculosis and AIDS must be strengthened in Chad. Mores cooperation is needed between tuberculosis and AIDS control programs. Emphasis should be placed on screening for tuberculosis, early tuberculosis treatment and diagnosis of HIV in tuberculosis patients. PMID- 12731313 TI - [Everything (or almost everything) about the Kappa coefficient]. AB - Sensitivity, specificity, and predictive value are the standard parameters used to evaluate the efficacy of diagnostic tests. However all three parameters assume the existence of a gold standard test. Thus the problem arises as to what method to use in the absence of a benchmark. The solution involves assessment of agreement between two raters. This article based on an example drawn from an African setting describes the value and computation of the most widely used tool for assessing agreement and reproducibility of measures, i.e., the kappa coefficient. PMID- 12731314 TI - [Clinical overview of New World tegumentary leishmaniasis]. AB - This richly illustrated article (80 color photographs) based on the authors' experience in French Guyana documents the clinical diversity of American tegumentary leishmaniasis. Main highlights include the often outstanding aspect of lesions, the high frequency of forms not associated with ulceration or scab formation that must be recognized to achieve diagnosis in travellers returning from endemic zones, and the special prognosis of clinical forms associated with intradermic, lymphatic or hematogenous spread. The article also reviews an original diagnostic method based on culture of cutaneous biopsy specimens on specific nutrient mediums that provides isolates in a high percentage of cases (80%) and thus allows identification of offending parasite. PMID- 12731315 TI - [Malaria treatment in children. 2. Severe malaria]. AB - Severe forms of Plasmodium falciparum malaria are one of the world's leading causes of infection-related death in children. The World Health Organization (WHO) has defined a set of severity criteria to improve diagnosis and speed antimalarial treatment. Although the pertinence of these criteria has not been documented in France, child travelers presenting such features require hospitalization in intensive care. The gold standard therapy is intravenous administration of quinine. According WHO recommendations, quinine therapy should begin with a loading dose barring contraindications. However French recommendations do not include the loading dose due to potentially dangerous side effects in young children and lack of proven life-saving effect. Artemisinin derivatives have been shown to be as effective as quinine and are increasingly used in endemic zones due to good tolerance and convenience of use. However due to concerns about neurotoxicity, artemisinin derivatives are rarely used in France, except in patients with contraindications or resistance to quinine. Management of specific complications is also necessary to reduce the high mortality of severe malaria, even in Western countries, and to prevent neurological damage. PMID- 12731316 TI - [Travel from women's perspective]. PMID- 12731317 TI - [Cerebellar disease in infectious mononucleosis]. PMID- 12731318 TI - [Obstructive trifid ureter (a case report)]. PMID- 12731319 TI - [Laryngeal cancer in women. Report of 3 cases in Congo]. PMID- 12731320 TI - [Lichen sclerosus et atrophicus of the penis: a precancerous lesion]. PMID- 12731321 TI - [Techniques and current practice of urodynamics. Problems and traps]. AB - Development of urodynamics particularly during the past decade are highlighted, as well as a number of issues to be faced in the near future: research into the etiopathogenesis of different conditions, finding more sensitive and specific diagnostic procedures to overcome the current ones, establishing more comprehensive indications for urodynamics examinations and, as a result of all the above achieving greater improvement of certain surgical procedures. Both the technique and interpretation of the different urodynamic examinations, as well as the more recent innovations, implementation issues and controversies are detailed to a highly up-to-date level. Neuromodulation and ambulatory urodynamics deserve thoughtful consideration in this paper. Finally, attention is given to the controversies and future challenges such as urodynamic research providing accurate diagnosis of lower urinary tract obstruction in women, establishing the indication of urodynamic studies in women with urinary exertional incontinence, outlining the indications of neuromodulation and ambulatory urodynamics, applicability of artificial intelligence systems, improvement of artificial sphincter materials, tissue growth for bladder enlargement and actual prevention of myelodysplasia. PMID- 12731323 TI - [Corrective surgery of penile inward curvature]. AB - OBJECTIVE: To evaluate the corporoplasty techniques performed on 59 patients to correct congenital penile curvature and Peyronie's disease. PATIENTS AND METHOD: From april 1991 to december 2001, 39 Nesbit technique, 12 Ruiz-Castane method and 8 Essed-Schroeder procedure were performed on forty-four congenital curvatures and fifteen Peyronie's diseases. The mean age was 24 years old. Trying to hide the knots, in all patients except five a nonabsorbable inverting sutures were placed. RESULTS: The mean follow-up was 12 months. Postoperatively, 53 patients (86%) had satisfactory cosmetic and functional results. Four patients of five with absorbable sutures had residual curvature which needed another successful surgical correction. One patient complained of penile shortening, one of glans hypoaesthesia, one of foreskin oedema and five of palpable plication sutures. No differences were found depending on the applied surgical technique. CONCLUSIONS: The results reported are in accordance with the literature showing a higher recurrence rate with the absorbable sutures. The outcome is very similar with the three described techniques. PMID- 12731322 TI - [Surgical management and complications of urinary stress incontinence: our experience in 385 patients operated on during the last 25 years]. AB - We present the results and complications found and compared in 2 large series of patients who underwent a surgical procedure to avoid Urinary Stress Incontinence in our hospital. From january of 1994 to december of 2001 we collected 195 patients with an average age of 57.9 y we compared them with a prior series of 189 patients collected between january of 1976 and december of 1993, with an average age of 53.6. The continence rate was similar in both series (70.8% pre-94 and 74.5% post 94), slight incontinence 22.7% pre-94 and 19.2% post-94 and for complete incontinence 6.3% pre-94 and 6.2% post-94. The Burch procedure was the most common and also shows the greatest effectiveness in our trial involving 113 cases and a continence rate of 74.3% in the first series and 144 cases and 77% respectively in the second series. The most common complications were very similar in both groups: tract urinary infection (5.2% and 9% first and second group), surgical wound infection (4.2% and 2.2% respectively). We conclude that in spite of the introduction of new and useful surgical procedure (TVT, periurethal substances injection...) the greatest long term success rate still corresponds to the Burch colposuspension. PMID- 12731324 TI - [Use of statistical analysis in original articles in "Actas Urologicas Espanolas." Accessibility for the reader]. AB - OBJECTIVES: To describe the use of statistical methods employed in original articles published in Actas Urologicas Espanolas as the first step to quantify the accessability for the readers. METHODS: Observational, retrospective, descriptive and transversal study to analize all the original articles published in Actas Urologicas Espanolas during the year 2000. Once the anectotic cases were excluded, 55 original studies were selected. In each article it was minuciously reviewed the methods and results sections, tables and figures included, to identify the statistical analysis used and to classify them into 18 categories with three accessability levels as Emerson and Colditz and Mora y cols, previously reported: descriptive statistics, bivariables analysis and complex analysis. A randomized sample of the originals were reviewed again by the same investigator three months later to evaluate the criteria liability. The accessability was defined as a) article dependent (rate of originals that readers with different statistical knowledge are able to understand) and b) analysis dependent (rate of statistical methods respect to the total performed overall the articles that readers are able to understand). RESULTS: Our major findings are that the more frequently used technics are descriptive analysis (39.3%), bivariable tables (12.1%), survival analysis (10%), t Student and z tests (6%), and nonparametric tests (8.6%). The accessability for a reader which statistical konowledge includes bivariable methods is 63.6% (IC 95% 49%-76%) and 79% (IC 95% 71%-85), article and analysis dependent respectively, rates that are similar to the reported for biomedical journals in our country. CONCLUSIONS: A great percentage of original articles in Actas Urologicas Espanolas includes complex analysis. It could be good that readers were able to increase our statistical and methodological knowledge to perform a critical approach to our publication. PMID- 12731325 TI - [The role of tumor markers in urologic consultation for screening, diagnosis, and follow-up of bladder cancer]. AB - PURPOSE: Review the tumor markers for screening and diagnosis of transitional cell carcinoma of the bladder. METHODS: The published literature on bladder cancer markers was identified using a MEDLINE search and critically analyzed. The sensitivity and specificity of the various markers were compared. RESULTS: Conventional non-invasive methods for diagnosis (symptoms and urine cytology) and for screening and follow-up of transitional cell carcinoma of the bladder are not very sensitive and must not be considered sufficient. Some tumor markers seem to be a good alternative to the routine urinary cytology. CONCLUSION: The role of tumor markers for screening and follow-up of bladder tumors as an alternative to cytology deserve consideration. In the future, more sensitive and specific markers may replace routine cystoscopy. PMID- 12731326 TI - [Major ambulatory surgery in urology. 5 years' experience]. AB - INTRODUCTION: The ambulatory surgery includes those surgical procedures that require a limited period of post-operative recovery, so that patients will be discharged from hospital on the same day of their surgical operation. OBJECTIVES: This publication aims at both, explaining our Service way of working as an integrated unit of ambulatory surgery, and it also tries to revise the results which have been obtained during our first 5 years working together. METHOD AND MATERIALS: Three hundred and thirty-nine patients, with an average age of 37.5 years (3-84), have been included in this report which sets out the medical record of this patients selection, their pathologies, the different procedures used, the kind of anaesthesia as well as the criterion to discharge them from hospital. RESULTS: Intra-surgical complications have arisen in 4 (1.2%) of our patients, immediate complications in 24 patients (7%) and late ones in 33 (9.7%) of our patients. According to the results of an anonymous inquiry which has been polled systematically to all our patients, 95% of those polled are completely satisfied with this new way of medical attention. CONCLUSIONS: The ambulatory surgery means a slight change in our patients' way of life. It also diminishes hospital costs, morbidity is similar to the one produced in hospitalization and it implies an important challenge for the professionals of this Service. PMID- 12731327 TI - [Unusual bladder tumors: primary epidermoid carcinoma, adenocarcinoma, and sarcoma. Clinical behavior. Our experience]. AB - INTRODUCTION AND OBJECTIVES: Non-transitional cell tumours of the bladder are both a diagnostic challenge for the pathologist and a therapeutic challenge for the urologist, because although uncommon (less than 5% of all malignancies of the bladder) they show different biological behaviours each requiring a unique approach. The most significant pathoanatomical types are: primary epidermoid carcinoma, primary adenocarcinoma and primary sarcoma. This paper presents an analysis of our experience in these types of tumours. MATERIAL AND METHODS: A retrospective study of unusual cases of cancer of the bladder seen in our hospital between 1988-2001. Their biological behaviour and the therapies applied are analysed. The extravesical origin of the tumour was ruled out in all cases. RESULTS: We found 21 cases of the following pathoanatomical varieties: 13 epidermoid carcinomas, 7 adenocarcinomas (3 urachal) and 1 sarcoma. Mean age was very different between epidermoid carcinomas (69.2 +/- 9.1 years) and adenocarcinomas (50.3 +/- 8.1). Tumour behaviour was very aggressive in all three histological varieties. At the time of diagnosis 19 patients had locally advanced stages (> or = T2). Although elective therapy was cystectomy, this was only feasible in 10 cases. Systemic chemotherapy (most frequently M-VAC) and/or local radiotherapy was used in 7 cases. 14 patients have died after a mean survival of 15.7 +/- 11.6 months. CONCLUSIONS: Late diagnosis of these tumours and their aggressive biological behaviour involve a gloomy prognosis. Only early diagnosis and radical therapy could improve the prognosis. PMID- 12731328 TI - [Multivariate analysis of recurrence and progression in stage T1 transitional cell carcinoma of the bladder. Prognostic value of p53 and Ki67]. AB - OBJECTIVE: To establish prognostic factors of recurrence and progression in stage T1 transitional cell bladder carcinoma, paying special attention to prognostic value of p53 and ki67. MATERIALS AND METHODS: 175 patients with incident bladder tumour at T1 category. The immunohistochemical study was performed using the monoclonal antibodies DO-7, for p53 and MIB-1, for ki67. Kaplan-Meier methodology was used, and a multivariate analysis using Cox's proportional hazards model was carried out in order to determine the variables associated with recurrence and progression. ROC curves were also drawn up. RESULTS: The average follow up period was 8.55 years. 5 and 12-year recurrence-free survival rates were 57.98% and 48.47%. The independent variables for recurrence were histological grade, tumour multifocality, tumour size > 3 cm, presence of carcinoma in situ and ki67 expression. 5 and 12-year progression-free survival rates were 83.12% and 75.63%. The independent variables for progression were age, tumour multifocality, solid microscopic morphology, p53 expression and ki67 expression. CONCLUSIONS: Solid microscopic pattern and p53 expression are the variables which best predict progression. A positive relationship was observed between p53 and progression: the greater the expression of p53, the greater the progression. Tumour multifocality and ki67 expression > 27% are the main prognostic factors for recurrence. PMID- 12731329 TI - [Mucinous adenocarcinoma of the urachus]. AB - OBJECTIVE: The Adenocarcinoma of the Urachus is very rare tumor, with an incidence of 1/5,000,000 inhabitants, represents less than 0.001 of all types of bladder cancer. CASE REPORT: A 51 year old man with a chronic history of suprapubic pain and hematuria. Physical examination and excretory urography were normal. The cystoscopy demonstrated a oedematosa area in cupola of bladder wall. The transuretral biopsy was moderately differentiated adenocarcinoma, with positive antibody to CK7 and CK20, the carcinoembryonic antigen was 6.6 ng/ml. Extended partial cystectomy was done, followed for chemotherapy and radiotherapy. CONCLUSIONS: The treatment of adenocarcinoma of the urachus with a combination of extended partial cystectomy, chemotherapy and radiation, is a effective treatment. PMID- 12731330 TI - [Diagnostic and therapeutic approach to bilateral germinal tumor. Presentation of a case and review of the literature]. AB - The germinal bilateral tumor represents from 1 to 4% of the testicle tumors, and it usually turns to be one of the most difficult ones when talking about diagnosis and treatment. We are attaching a 30 years old patient case, with a diagnosis consisting in a seminoma with yolk sac tumor area from the left testicle, that was treated with orquiectomy. Eleven months later, the patient presented a second neoplasm in the contralateral testicle, whose histology was a classical seminoma with carcinoma "in situ" peritumoral, being also treated with orquiectomy. Medical literature is revised with emphasis in the involved risk factors and the techniques used to treat these testis tumor groups: carcinoma "in situ" detection appears to be the strongest predicting factor. Testicular contralateral biopsy is proposed in some select cases. Even though radical orquiectomy is still the treatment to choose, local radiotherapy and conservative surgery are also an option in patients with bilateral tumor. PMID- 12731331 TI - [Melanoma of the penis]. AB - We present a case of penile malignant melanoma. Its rarity of less than 1% of masculine genital malignant neoplasms, the absence of standardised protocols as for other neoplasms, impel us to its publication. PMID- 12731332 TI - [Necrosis of the skin of the penis as a complication of the use of a urine collector]. AB - We illustrate a case of penile skin necrosis in a patient suffering from urinary incontinence caused by a secondary neurogenic bladder that, in turn, results from a spinal cord injury. The skin necrosis developed out of continuous pressure from the condom catheter. We report the case bearing into consideration that these complications are extremely rare and that references in the literature to the topic are likewise scarce. However, due to the high frequency of patients suffering from incontinence who use these devices, we believe it necessary to account for the possible consequences which might derive from an incorrect use of them. PMID- 12731333 TI - [Bladder malacoplakia with lymphatic involvement and an aggressive course]. AB - Malacoplakia is an unusual chronical granulomatous disorder. In the urinary tract is more frequent in female, in 75 percent of cases involve the bladder. Pelvis extension of this disease is infrequent, and even less frequent is the involvement of pelvic and retroperitoneal lymph nodes. We present the second case in the literature of bladder malacoplakia with extravesical and pelvic node involvement. PMID- 12731335 TI - [Closure of laparoscopic access ports using a Reverdin needle: a new use for an old instrument]. PMID- 12731334 TI - [Association of renal carcinoma and the exposure to ionizing radiation after the Chernobyl accident]. AB - After the nuclear accident of Chernobyl, in the population of zones contaminated the malignant renal tumors was increased from 4.7 to 7.5 per 100,000 of total population. Cesium 137 (137Cs) constitutes 80-90% of the internal exposure of these people as well as eliminated through kidneys becomes an important risk factor. We present a case of a patient, residing in radiocontamined area, who consulted for abdominal pain and left flank mass. We review relevant literature and the management of these patients. PMID- 12731337 TI - [Effectiveness of exercise programs in lipid metabolism disorders]. AB - Five-six hours per week regular physical activity with the large muscle groups, amounting more than 2000 Kal per week, utilizing 50-70 percent of the aerobic capacity or 60-85 percent of the heart rate reserve, increases the HDL cholesterol level, decreases the triglyceride. Trainings thorough months and years decreases the LDL-cholesterol and total cholesterol levels also. Magnitude of the changes are the same (HDL) or comparable to the effects of the lipid decreasing medicaments. Beyond the beneficial effects on the plasma lipids, the adequate physical activity has a unique influence on the cardiac and cardiovascular functions, on the carbohydrate metabolism, on the blood pressure, on the mood and psychological status. PMID- 12731336 TI - [Cellular and molecular changes of the liver in sepsis and in systemic inflammatory response syndrome (SIRS)]. AB - This paper provides a review on the changes in the molecular and cellular level of the liver during sepsis and systemic immune response syndrome. The different function of the various liver cells and their mediators are analyzed. Dual role of nitric-oxide and carbon-monoxide are discussed followed by an overview about acute phase proteins and heat shock proteins in septic liver. A detailed description is presented on the role of the Toll-like receptors and their signaling pathways in the liver during sepsis and systemic immune response syndrome. Finally, the possible mechanisms leading to apoptosis of the liver cells are shown and the most recent therapeutical challenges are discussed. PMID- 12731338 TI - [Antilipemic therapy and rhabdomyolysis]. AB - The authors review the pathomechanism and frequency of side effects (myopathy and rhabdomyolysis) of the lipid lowering agents, statins and fibrates, on the basis of large prospective studies. They review in detail the possible pathomechanism of myopathy as caused by statins and fibrates. The cytochrome P450 enzyme system plays a role in the interaction of other drugs with long term statin therapy. The long term administration of statin and fibrate to patients with primary and secondary hyperlipidemia was considered safe on the basis of our experience and literature data. The combination of statin and fibrate is also considered safe and useful in the treatment of combined hyperlipidemia. As a precautionary measure, we emphasize the ruling out of liver, kidney and muscular diseases in such patients. Lipid lowering therapy is efficient in the primary and secondary prevention of cardiovascular diseases if applied cautiously. PMID- 12731339 TI - [The role of medical schools in progressive health care]. AB - AIM: To analyse the predicted and observed role of medical schools in the progressive health care. DATA AND METHODS: The data derives from the financial database of the National Health Insurance Fund (NHIF) and covering the period 1998-2001. In this study the authors calculated the market share of the medical schools within the financing of the NHIF in the field of out- and in-patient care, renal dialysis, CT/MRI examinations, task financed services under special rules and fee for progressive (tertier level) care. The authors performed a detailed analysis concerning the Diagnosis Related Groups (DRG) of active inpatient care, where the market share of medical schools was calculated within the TOP-15 DRGs with highest and lowest point value, and within the TOP-15 most common and most infrequent DRGs. RESULTS: The market share of medical schools increased from 13.7% (1998) to 15.0% (2001). The increase was significant in the active and chronic in-patient care and in renal dialysis. The market share of medical schools from progressive fee decreased from 32.3% (1998) to 26.5% (2001). Within the active in-patient care the medical schools provide health care for 37.7% of the most difficult cases with highest DRG value and for 30.1% of the patients with rare diseases. The market share of the medical schools is much lower in cases with lowest DRG value (10.2%) and in most common cases (9.8%). CONCLUSION: The medical schools fulfilled their expected role in the progressive (tertier) care, answering the public and professional expectations. PMID- 12731340 TI - [Eosinophilic gastritis simulating gastric carcinoma]. AB - A case of eosinophilic gastritis is reported in a 65-year-old woman. The disease presented with signs of delayed emptying, vomiting and substantial thickening of the antrum. Partial gastrectomy was performed because of the suspicion of gastric cancer and pyloric stenosis. Histopathology established the diagnosis of eosinophilic gastritis of mural type. Clinicopathological features of the disease, as well as its differential diagnosis are summarised in the discussion. PMID- 12731341 TI - [Quo vadis, pathologia?]. PMID- 12731344 TI - [Psychosurgery, a different approach to psychiatric disease]. PMID- 12731345 TI - [Adaptation to work time reduction in public hospitals]. PMID- 12731346 TI - ["Silence" on suicide. Interview by Christian Bourdeux]. PMID- 12731347 TI - [Pathological bonds, between the ideal and imaginary family]. PMID- 12731348 TI - [From therapy to family therapeutic care]. PMID- 12731349 TI - [Symbolic adoption, affiliation and identification]. PMID- 12731350 TI - [From placement to care]. PMID- 12731351 TI - [Success and weakness in therapeutic family care]. PMID- 12731352 TI - ["The child's response is different and we are working for that"]. PMID- 12731353 TI - [Nurse in a medical-psychological center, an identity in construction]. PMID- 12731354 TI - [To the memory of Fedor Andreeevich Serbinenko]. PMID- 12731355 TI - [Experience with endovascular occlusion of cerebrovascular aneurysms by means of microspirals]. AB - The paper summarizes the outcomes of endovascular treatment of 42 patients (19 males and 23 females) aged 24 to 68 years who had cerebrovascular aneurysms, 7 of them were operated on in the acute phase of subarachnoidal hemorrhage. There were small [n = 27 (61.4%)], large [n = 14 (31.8%)], and giant [n = 3 (6.8%)] aneurysms. According to their site, aneurysms were found in the internal carotid [n = 22 (50%)], the middle cerebral artery [n = 2 (4.5%)], the anterior cerebral artery-anterior communicating artery [n = 10 (22.7%)], the basilar artery [n = 9 (20.5%)], and the vertebral artery [n = 1 (2.3%)]. MDS tungsten and platinum miscospirals (Balt, France) were employed for aneurysmal occlusion. Twenty seven (61.4%) aneurysms were totally and subtotally closed by means of microspirals. Partial occlusion of the aneurysmal cavity was achieved in 12 (27.2%) cases. There were poor results associated with spiral migration into the carrying vessel in 5 patients: an increase in aneurysmal volumes occurred in 3 patients and in 2 patients with partially occluded giant aneurysms within up to a year. Three patients died. Angiographical control was exercised in 12 patients at months 6 to 18. The analysis of long-term results indicated that the aneurysms initially totally closed failed to be recanalized later on (8 patients). With subtotal or partial occlusion, there was a change in the position of spiral coils in the aneurysmal cavity, namely, their displacement to the bottom and an increase in the dimension of a functioning aneurysmal part (2 patients). The use of microspirals for occlusion of cerebrovascular aneurysms is a low-traumatic and effective treatment. The best results were obtained with the occlusion of small and large aneurysms with their well-defined neck. PMID- 12731356 TI - [Embolization of cerebrovascular aneurysms with controlled microspirals (complications and technical problems)]. AB - The paper summarizes five-year experience with mechanically detachable microspirals for embolization of 62 arterial aneurysms in 59 patients. The complications and technical problems associated with both the specific features of this technology and anatomic types of the structure of vessels and aneurysms are described. Methods for the prevention and treatment of these complications are proposed. PMID- 12731357 TI - [Main anastomoses of external carotid artery branches with the beds of the internal carotid and the vertebral artery and a role of pharmacological tests in the determination of their functional value at tumor embolization]. AB - In 21 patients with meningovascular tumors (meningiomas, angiofibromas, etc.), 53 vessels were tested during preoperative embolization of extracranial tumors. Two groups of drugs were superselectively intraarterially administered in turn--first barbiturates, such as thiopental sodium, 15-20 mg in 2 ml of sodium chloride solution, to detect anastomoses with cerebral vessels, then local anesthetics (lidocaine, 10-20 mg in 2 ml of sodium chloride solution, to reveal the vessels feeding cranial nerves. If drug tests were negative, the afferent was embolized with under 300 microns particles. If they were positive this was done with only large particles (above 300 microns). Injection of thiopental sodium into the occipital artery provided a positive result in 1 (1.8%) of the 53 cases. There were no complications after negative tests, followed by embolization. Drug provocative tests during embolization of the vessels feeding extra- and intracranial tumors make it possible to reduce the risk for persistent ischemic events associated with the entry of artificial emboli into the cerebral vessels and vasa nervorum and to conduct the optimum embolization of afferents. PMID- 12731358 TI - [Early and late results of the intravascular treatment of cerebral aneurysms with a detachable balloon-catheter]. AB - The paper analyzes the outcomes of intravascular treatment in 87 patients with cerebral aneurysms with separable a balloon catheter with a valve device. In the cold and hemorrhagic periods, mortality rates were 6.4 and 40%, respectively. The postoperative follow-ups lasted 1 to 17 years (mean 7.7 years). Recurrent subarachnoidal hemorrhage (SAH) was noted in 6.5% of cases, with not earlier than 4.5 years after intervention. The remaining patients (even with partial aneurysmal occlusion) had no recurrent SAH. PMID- 12731359 TI - [X-ray contrast filler for balloons at endovascular surgery (20 years of experience with original balloon filler)]. AB - In 1984, the authors designed a X-ray contrast balloon filler polyacryl amide gel (PAAG) that has a polymerization rate of 1-3 min. PAAG has been used to fill over 5000 balloons for nearly 20 years. With this filler, recurrent anastomoses reduced by 1.5 times as compared to previously used silicone. PAAG is a reliable and easy-to-use balloon filler. Its use as a balloon filler has been permitted by the USSR Ministry of Health. PMID- 12731360 TI - [Temporal arterial clipping and autoregulation of cerebral blood flow at surgery for intracranial aneurysms]. AB - Preoperative dynamic autoregulatory responses in 20 patients considered as candidates for vessel temporal clipping (TC) and/or arterial hypotension (AH) at surgery for aneurysms were studied by the thigh cuff method. Normal autoregulatory responses at a regulation rate (RoR) of 20.4 +/- 2.8%/s were found in 10 patients > 30 days after subarachnoidal hemorrhage (SH) while RoR was 12.5 +/- 2.5%/s in 10 other patients (30 days, grade III WFNS showed autoregulation impairment. TC and/or AH were proceeded in 17 operated patients. One postoperative death was due to pulmonary embolism. Two poor results with persistent neurological disorders at discharge were associated with TC but in 1 patient. This patient had severe vasospasm and the lowest RoR (9%/s). There were no TC and/or AH postoperative complications in other 14 patients. All of them and 3 other patients with direct aneurysm clipping had a fair outcome. PMID- 12731361 TI - [Use of different endovascular treatments for arteriovenous malformations of the spinal cord]. AB - The paper presents outcomes of treatment of patients with arteriovenous malformations (AVM) of the spinal cord by using endovascular techniques in 171 patients. There were a total of 364 occluded afferent vessels involved in the blood supply to and development of AVM. A classification of spinal cord AVM is presented. Methods for occlusion were described in relation to the type AVM. Malformations were thrombosed with polyvinyl acetate (PVA) emboli in 129 patients; balloon occlusion of afferent vessels was performed in 6, as described by F. A. Serbinenko; histoacryl glue was applied in 27 cases. A combined method of occlusion of afferent vessels was used with PVA emboli and balloons in 9 patients. The clinical picture of spinal cord AVM was outlined. Emphasis is laid on diagnostic methods, such as selective angiography and magnetic resonance imaging, as well as on their role in postoperative control studies, in determining the degree of thrombosis, the presence of ischemic events, and subarachnoidal space patency. Endovascular treatment used in patients with AVM at different levels of the spinal cord alleviated neurological symptoms in 155 (91%) patients. There was no improvement in 12 (7%) patients. Complications at angiography and embolization were observed with deteriorated neurological symptoms in 4 (2%) cases. Further development and improvement of endovascular treatments in patients with spinal cord will be promising if new intervention technologies are introduced in clinical practice. PMID- 12731362 TI - [Formation of intervention neuroradiology at the department of neurosurgery of the military medical academy]. PMID- 12731363 TI - Simplifying your menu. PMID- 12731364 TI - Dietitians in small hospitals: they must wear many hats. PMID- 12731365 TI - Management accountability. PMID- 12731366 TI - [Surgical treatment of Valsalva sinus aneurysms and fistulas]. AB - INTRODUCTION: Valsalva sinus aneurysms and fistulas (ASV; FSV) are rare aortic pathologies and present in only a few percent of patients with aorta and/or left arterial orifice disease. Etiology of ASV and FVS is congenital and acquired. MATERIALS AND METHODS: 40 patients (pts) were operated on in the Department of Cardiovascular Surgery and Transplantology from 1978 to 2002 due to pathology of Valsalva sinus (< 1% of all operations in ECC). 18 pts were diagnosed with Valsalva sinus aneurysm and 22 with its fistula. Inflammatory etiology was confirmed in 22 pts (5%). Most common pathology was localized in the right and/or coronary lacking Valsalva sinus (30 pts-75%). FSV penetrated into the right atrium or ventricle in 20 pts (91%). Among accompanying pathologies aortic valve insufficiency was most frequently found. In 80% of pts with ASV aortic allografts were implanted. In 70% of pts with FSV, fistula was sutured with interrupted pledgeted mattress sutures. Accompanying anomalies were corrected simultaneously. Mechanical aortic valve was implanted in 9 pts. RESULTS: 2 pts died in the early post-operative period and 3 in the follow up. Reoperation (with good results) was performed in 4 pts. At discharge all pts were in NYHA I/II except 1 pt in NYHA III. Objective estimation of patient's cardiopulmonary sufficiency did not differ from the subjective one. 24 pts performed normal life activities after operation. The echocardiography confirmed effectiveness of surgical treatment. CONCLUSION: 1. ASV and FSV require different surgical techniques. 2. Surgical treatment efficiently reduced symptoms. 3. Allografts were found very useful. 4. Accompanied anomalies should be treated simultaneously. PMID- 12731367 TI - [Hyperhomocysteinemia and advancement of atherosclerosis in patients with chronic renal failure on maintenance hemodialysis]. AB - Cardiovascular diseases connected with atherosclerosis are the main factor of morbidity and mortality in patients with end-stage renal failure. Hyperhomocysteinemia is a known and independent risk factor of atherosclerosis, occurring in 85-95% patients treated with hemodialysis. The aim of this study was to analyse relation between plasma level of homocysteine and chosen indicators of atherosclerosis development and also examined retrospectively cardiovascular complications in these patients. The study was carried out in 100 patients on hemodialysis who were divided into two groups: 72 patients with mild (20.74 mumol/l +/- 3.75) and 28 patients with moderate hyperhomocysteinemia (38.81 mumol/l +/- 9.81). Ultrasonographic examinations of Carotid Communis Artery Intima-Media Thickness (IMT), Ankle-Arm Blood Pressure Index (AABPI), echocardiographic parameters and biochemical examinations such as: PTH, folic acid and Vitamin B12, total protein, albumin, fibrinogen, glucose, total, LDL and HDL cholesterol, transferring, apolipoprotein B, lipoprotein (a), sodium potassium, calcium, phosphate, magnesium, iron, ferritin, urea, creatinine, uric acid and value of Hb, Ht, total iron binding capacity and transferring saturation, were performed. Patients with hypertension were divided into groups according to the number of taken anti-hypertensive drugs. Hyperhomocysteinemia was confirmed in 96% of patients. Frequency and type of acute cardiovascular complications were not related with the level of hyperhomocysteinemia. Statistically significant difference between IMT and level of hyperhomocysteinemia was observed. In patients with mild hyperhomocysteinemia IMT was 0.68 mm +/- 0.24 whereas in patients with moderate hyperhomocysteinemia 0.80 mm +/- 0.25, p < 0.036). Positive correlation between level of homocysteine and IMT (r = 0.22, p < 0.03) was noted. Based on this study, we concluded, that measurement of intima-media thickness is a good indicator of atherosclerosis development and correlates with hyperhomocysteinemia in patients on maintenance hemodialysis. It clearly confirms the role of hyperhomocysteinemia as significant risk factor of atherosclerosis in those patients. PMID- 12731368 TI - [Cutaneous changes occurring after taking non-steroidal anti-inflammatory drugs- five-years' retrospective studies]. AB - In the last years we can observe an increase incidence of dermatoses after drug administration, which later require specialistic help and hospitalization. It especially concerns with nonsteroidal, anti-inflammatory drugs (considering their widespread application, possibility of purchasing them without prescription and large media advertising. The authors of this article wanted to present the significance of this problem analysing a group of patients at the Department of Dermatology in Cracow in the years 1997-2001. Attention was paid to a variety of cutaneous changes after they had been treated with nonsteroidal anti-inflammatory drugs (NSAID). The dermatoses which are not commonly considered as drug related, have also been presented. In general, medical documentation of 193 patients being suspected of having symptoms of drug related character of cutaneous changes were presented. The given drug was recognized as being the cause of disease development based on the method recommended by the French Surveillance Network. In 31 patients a connection of the cutaneous changes with NSAID administration was proved. In the analysed group of patients, women dominated (61.3%), the mean age of the examined patients was 42 years. The increase in occurrence frequency of dermatoses after taking NSAID was observed in regard to all the hospitalized patients (from 0.48% in the year 1997 to 0.99% in the year 2001). From among the drugs responsible for causing cutaneous changes which required hospitalization, aspirin took the first place, pyralgin the second, and paracetamol the third one. Urticaria prevailed in cutaneous changes after having taken NSAID. More rarely other cutaneous changes, like erythema multiforme or drug induced exanthema, took place. The fact that various drugs often cause similar cutaneous changes and even the same drug can provoke different morphological reactions makes the diagnostic process of the described dermatoses extremely difficult. PMID- 12731369 TI - [Esophageal scintigraphy in patients with systemic sclerosis: clinical symptoms correlated with the esophagus noted by the patients]. AB - Systemic sclerosis (SSc) is a multisystem connective tissue disorder characterized by excessive accumulation of the extracellular matrix. About 90% of the patients with SSc have detectable gastrointestinal tract involvement, 50% of these patients have clinically important manifestations. The aim of the study was estimation of the connection between subjective symptoms from the esophagus and results of the esophageal scintigraphy. 30 patients with systemic sclerosis were comprised to the study. Esophageal scintigraphy was performed in each patients, as well as questionnaire about type, intensity and frequency of esophageal dysmotility. Also it estimated was stiffness of the skin according to modified Rodnan score. Independent of esophageal dysmotility symptoms which were noted by patients, esophageal scintigraphy registered changes in 97% of the patients. The most often esophageal dysmotility was observed in lower part of the esophagus. Statistically Dysmolity of the central portion of the esophagus was observed more often in patients with diagnosed ISSc. PMID- 12731370 TI - [Are there relationship between Helicobacter pylori infection and gastrin levels in long term hemodialysed patients?]. AB - Helicobacter pylori (H. pylori) infection in patients with chronic renal failure plays an important role in the pathogenesis of upper gastrointestinal tract diseases. In our earlier study we did not find significant relationship between parathormone (PTH) concentration abnormalities and H. pylori infection in hemodialysis patients (HD pts). The aim of our present study was to examine other parameters and interrelationships between them, connected with H. pylori infection in HD pts. The serum concentration of the following substances were measured: gastrin (G), parathormone (PTH), and IgG antibodies against H. pylori. The study was conducted in 65 (36 M, 29 F) stable HD pts: age 49 +/- 12 years, dialysed from 6 to 288 months. The control group (CG) consisted of 15 healthy people, mean age 38 +/- 8 years. The mean concentration of serum PTH serum was significantly higher in HD pts than in CG (538 +/- 520 vs 35 +/- 9.3 pg/ml, p < 0.001). The mean concentration of gastrin in serum was significantly higher in HD pts than in CG (155.37 +/- 235.4 vs 87.97 +/- 9.5 pg/ml, p < 0.01). The mean concentrations of IgG antibodies against IgG H. pylori was similar in HD pts and CG (88.1 +/- 80.0 vs 91.34 +/- 66.8 U/ml). We found significant positive correlation between IgG antibodies against H. pylori infection and gastrin level in serum of HD pts (r = 0.315, p < 0.001). We did not find significant correlation between concentrations: PTH vs G and PTH vs IgG against H. pylori. CONCLUSION: Helicobacter pylori infection contributes to hypergastrinaemia in hemodialysed patients. There is no relationships between PTH abnormalities and H. pylori infection. PMID- 12731371 TI - [Some parameters of platelet activation in patients with uremia before and after hemodialysis (HD)]. AB - Patients with uremia show a tendency for hemorrhagic diathesis and thrombi formation. These disorders have been attributed to functional abnormalities of platelet and their interaction with the vascular wall. The aim of the present study was to elucidate whether and how in patients with uremia, hemodialysis affects the level of platelet activation markers, i.e.: beta-TG and PF4, and whether the changes in the examined parameters depend on creatinine level and on the type dialysis membranes used during HD. The study involved 70 patients with uremia subjected to repeated hemodialysis and 70 healthy subjects. Prior to HD, we observed a decrease in PLT, and increase in beta-TG concentration and a drop in PF4 (p = 0.000), compared to control group. Immediately after the procedure a significant increase was observed in PLT and decrease in PF4 concentration, as well as a slight rise in beta-TG level, compared to the values obtained before HD. We also found statistically significant differences in beta-TG and PF4 concentrations prior to HD, irrespective of creatinine concentration, compared to control group. Only at creatinine concentration of 10.0-15.5 mg/dL, PLT differences significantly from PLT in control group. Following HD, creatinine dependent statistically significant changes were noted in PLT, in comparison to the values obtained before the procedure. The analysis of platelet parameters revealed no statistically significant differences associated with the type of the dialysis membrane used for HD. PMID- 12731372 TI - [Physical fitness of children after operation of ventricular septal defect of the heart]. AB - Physical fitness of 26 children aged 8-12 years, being 4-7 years after cardiosurgical correction of ventricular septal defect of the heart (VSD) was studied. The results were compared with the ones obtained in 23 healthy children of the same range of age. There were continuously registered in these children the gas exchange values with direct method using progressive exercise test. There were studied minute oxygen uptake, minute carbon dioxide production as well as minute ventilation. In the analysis of physical fitness the following parameters were used: total time of effort, maximal heart rate, maximal power, maximal oxygen pulse, restitution pulse ratio, maximal minute oxygen consumption and ventilatory anaerobi threshold (VAT) determined individually by V-slope method. Comparing both groups of children the statistically significant differences of the VAT values were observed. This evidences the lower physical fitness of children after cardiosurgical correction of VSD. PMID- 12731373 TI - [Developmental age idiopathic osteoporosis--diagnostics and rehabilitation]. AB - Developmental age idiopathic osteoporosis is a rare metabolic disease, which pathomechanism is different from involutional osteoporosis. Its etiology still remains unknown. An onset of the disease is rapid, affecting previously healthy children of both sexes aged 3.5-15. A rapid loss of bone mass is especially remarkable. Clinical picture of the disease includes pain localised initially in feet, then in thoraco-lumbar section of the vertebral column, gradually progressive deformation of the longitudinal axes of limbs, gait pathology and progressive depletion of a lower limb muscle strength. The onset of the disease is non-specific. Diagnosis is possible on the basis of roentgen imaging depicting rarefaction of cancellous layer of the bone, numerous infractions of long bones and typical for osteoporotic deformation of vertebral shafts with reduction of their vertical dimension. Current research presents two-staged method of rehabilitation applied in developmental age osteoporosis. The main principle of treatment is based on increasing axial load of lower limbs in antigravitational position causing joint surfaces to be pushed together. Final stage of rehabilitation includes interval training on a moving track. PMID- 12731374 TI - [Psychosocial and nutritional risk factors for cardiovascular disease- longitudinal cohort study. Assumptions, goals and methods of the screening survey]. AB - In Poland after two decades of increase in mortality due to Coronary Heart Disease (CHD), the trend reversed and CHD mortality has decreased since 1992. Dynamic change in CHD mortality reflects changes in the incidence and can not be explained by improvement in the acute medical care. Neither could it be explained by changes in the exposure to known cardiovascular disease risk factors i.e. smoking, hypercholesterolemia and hypertension. There was a coincidence of decreasing trend in CHD mortality with rapid political and socioeconomic reforms in the country, which influenced lifestyle and the psychosocial environment of the individuals. Results of the studies indicate that health inequalities could be partially explained by psychosocial factors. Other studies suggested that favorable changes in diet and in alcohol consumption could contribute to favorable changes in health of the Polish population. The long term goal of the present project is to assess the relationship between the risk of coronary heart disease and 1) psychosocial factors, 2) type of diet (consumption of fresh fruits and vegetables, and, 3) alcohol consumption. Assumptions and goals of the project as well as methods of the initial screening, in which a representative sample of men and women at age 45-64 years, residents of Cracow was examined, are described. PMID- 12731375 TI - [Psychosocial factors and self reported health in residents of Krakow at age 45 64]. AB - The aim of the paper was to find a relationship between self-reported health and psychosocial factors i.e. education level, civil status, job effort, job reward and general perceived control, perceived control over health and over live. The studied sample were 1154 men and 1156 women, randomly selected from population of Krakow aged 45-64 years. Data on self-reported health, smoking, perceived control, job effort and job reward was collected using standard questionnaire. After adjustment for age and sex, the risk of "bad" or "very bad" self-reported health was higher for those with elementary compared to university education (OR = 2.43, 95%CI = 1.72-3.44), and for those with secondary compared to university education (OR = 1.77, 95%CI = 1.24-2.53), and for high job effort (OR = 2.21, 95%CI = 1.33-3.67), low general perceived control (OR = 2.09, 95%CI = 1-61-2.71), low perceived control over live (OR = 2.12, 95%CI = 1.63-2.74) and low perceived control over health (OR = 1.92, 95%CI = 1.45-2.52). Being married was associated with lower risk of "bad" or "very bad" self-reported health (OR = 0.59, 95%CI = 0.44-0.79). Further adjustment to CVD risk factors, i.e. smoking, systolic blood pressure, blood total cholesterol did not change the results significantly. PMID- 12731376 TI - [The occurrence of atopic hypersensitivity in children with adenoid hypertrophy]. AB - Allergic sensitization of the airways occurs in the mucosa. The lymphatic structure closest to the nasal mucosa is the adenoid. Adenoid hypertrophy (AH) may cause significant morbidity in children but its relationship to atopy has not been intensively studied. To examine the influence of atopy on the adenoid hypertrophy we studied 134 children ages 4 to 8 years, with an average age of 5.4 who had AH. In the analysed group 102 children suffered from difficulty in nose breathing, 61 of children from recurrent upper respiratory tract infections, 51 from recurrent otitis and 40 from hearing disorders. On the basis of allergic and immunological examinations, in 30.6% atopy was confirmed--41 out of 134 children showed positive skin test for allergy, mostly to different kinds of pollen or house dust. From these positive patients the respective allergens could be determined by RAST tests in serum of 87%. Increased total IgE level in serum was confirmed only in 17.5%. 8.5% of children showed peripheral blood eosinophilia. Some children were qualified for adenotomy and removed adenoid was examined histopathologically. Pharmacological treatment was also recommended. CONCLUSIONS: The study shows that allergy and sensitivity to different kinds of allergens is an important risk factor for a greater degree of AH in children. Therefore, early prevention of exposure to them may help reduce occurrence of AH. PMID- 12731377 TI - [Specific-pollen immunotherapy in the treatment of oral allergy syndrome in patients with tree pollen hypersensitivity]. AB - Over the last three decades several authors have described the existence of an association between sensitivity to different pollens and sensitivity to diverse edible vegetables. Most patients with birch pollen allergy report oral allergy symptoms after eating fresh apples and other vegetable foods. Major birch pollen and apple allergens, Bet v 1 and Mal d 1, and other antigens are highly homologous; as a consequence, pollen-specific immunotherapy (SIT) might be expected to improve apple hypersensitivity. A prospective study was carried out in 27 birch pollen-sensitive patients with apple-induced OAS (oral allergy syndrome) who received injection immunotherapy for 24, or 36 months. Eight patients not submitted to SIT and followed up for 24-36 months were used as a control. Sixteen patients (59%) reported a significant reduction (37%) or a total disappearance (22%) of OAS symptoms after SIT. CONCLUSION: SIT with birch pollen extracts effectively reduces clinical apple sensitivity in some cases; these effects are not paralleled by a similar reduction of specific IgE and skin reactivity. PMID- 12731378 TI - [Current views of surgical procedures in the treatment of hypertrophic nasal conchae]. AB - Surgical treatment aiming at decreasing the size of the nasal conchae is controversial and it should be performed only after all alternative kinds of therapy have appeared unsuccessful. The incision of hypertrophic nasal conchae can also be performed together with the operation of the nasal septum. The excision of the entire concha is performed very infrequently. Usually, only the soft tissue of the concha is excised, but the excision of the hypertrophied tissue is most desirable. Some patients may undergo the cryotherapy. PMID- 12731379 TI - [Aquaporin water channels in water balance regulation in the kidney]. AB - The aquaporins (AQP) are a family of small transmembrane water channels. The discovery of AQP has provided insight into molecular mechanisms underlying renal water absorption and its regulation by vasopressin. Seven types of AQP have been identified in the kidney. AQP1 has been localized in the proximal tubule and descending thin limb, while AQP2, AQP3, and AQP4 are expressed in the collecting duct. Of these isoforms, AQP2 expression and intracellular trafficking is tightly regulated by vasopressin. Decreased expression of renal AQP has been detected in several disorders associated with polyuria and impaired ability to concentrate urine, as exemplified by nephrogenic diabetes insipidus or renal failure. In contrast, increased expression of AQP is seen in conditions leading to water retention, such as congestive heart failure, liver cirrhosis, and syndrome of inappropriate antidiuretic hormone secretion. Thus, the understanding of molecular structure and function of aquaporins may have important implications for therapy of water balance disorders. PMID- 12731380 TI - [Thyrotropin--TSH secreting pituitary tumor]. AB - Thyrotropin-releasing pituitary tumors represent 0.9 to 2.8% of all pituitary adenomas. They cause secondary or central hyperthyroidism. The diagnosis of these tumors has been increasing in the past 20 years. It was produced by introduction of the sensitive immunoradio-metric assay of TSH and better radiological imaging (magnetic resonance imaging). TSH--secreting pituitary adenomas are aggressive and invasive neoplasms. Most reports describe a poor outcome after pharmacological therapy, surgery and radiation therapy. Presently the diagnosis of thyrotropin-secreting pituitary tumor is based on the lack of: a. inhibition of TSH levels in the presence of increased free thyroid hormones; b. response of TSH to stimulation with TRH; c. and presence of a abnormal, neoplastic(adenomatous) intrasellar or parasellar mass. Surgical excision (selective adenomectomy) by the transsphenoidal route is the first treatment. Craniotomy should be reserved for parasellar tumors with significant lateral extension. Pharmacological pretreatment with long acting somatostatin analogues is recently a standard before surgery. This medical treatment of the TSH-omas is effective in reducing TSH and free thyroid hormone plasma levels. Administration of the somatostatin analogues causing tumor mass shrinkage and changes consistency. This pretreatment is effective therapy and improves surgical outcome especially in patients harbouring macroadenomas. Radiotherapy is noncurative and produces long term complications (hypopituitarism). Authors present and discuss current cure criteria of TSH-omas with reference to their clinical experience. PMID- 12731381 TI - [Leptin and thyroid hormones]. AB - Leptin, a satiety hormone is a protein produced by the adipose tissue that regulates appetite and energetic balance of the body. Results of investigations of serum leptin in patients with hyperthyroidism or hypothyroidism are controversial. Influence of thyroid hormones on leptin secretion is complex and partially unrecognized. PMID- 12731382 TI - [Evaluation of Cepan Cream after 15 years of treatment of burn scars]. AB - Cepan Cream is used for the topical treatment of scars and keloids resulting from burns, post-operative scars, and contractures. Cepan Cream makes scars more elastic, softer and paler. Plant extracts, heparin and allantoin in Cepan act on the biochemical processes in the developing connective tissue, preventing the formation of hyperplastic scars. These active ingredients enhance swelling, softening and loosening of connective tissue. It exerts softening and smoothing action on indurated and hyperplastic scar tissue, improving collagen structure. It promotes tissue regeneration and reduces exuberant granulation. Cepan is well tolerated. PMID- 12731383 TI - [Usefulness of ultraviolet radiation in dermatological treatment]. AB - Ultraviolet radiation causes a complex cascade of changes resulting in suppression of immune reactions. The anti-inflammatory and immunosuppressive properties are widely used in treatment of many dermatoses. Currently applied phototherapy methods are presented in this paper. The knowledge of the UV mechanisms, phototherapy schedules, appropriate indications and restrictions allows to achieve better therapeutic results and decreases the risk of side effects. PMID- 12731384 TI - [Nonfunctional pituitary adenoma and pulmonary sarcoidosis--a case report]. AB - A case of the coexistence of nonfunctional pituitary adenoma and pulmonary sarcoidosis is reported. 39 years old female presented symptoms of a pituitary gonadal axis insufficiency, visual deficit and dizziness. CT pituitary imaging revealed large intra and extrasellar tumour. Histological examination of the tissue obtained at transsphenoidal surgery showed chromophobic adenoma. Hypopituitarism and transient diabetes insipidus occurred after the surgery. The adequate replacement therapy with hydrocortisone and sex steroids was introduced. At the time of the pituitary tumour diagnosis enlargement of pulmonary lymph nodes was also observed. Based on histological examination of tracheal and bronchial epithelium specimens obtained during bronchoscopy the diagnosis of pulmonary sarcoidosis was made. No other systemic sarcoidosis localisation was confirmed. Histological re-evaluation of adenoma showed no noncaseating granuloma tissue. The overlapping symptoms of pituitary adenoma and other intrasellar masses may result in diagnostic difficulties, particularly in the presence of systemic disorders in which this gland may be involved. PMID- 12731385 TI - [Complete obliteration of false lumen in thoracic aorta after surgical treatment of acute type De Bakey I aortic dissection--a case report]. AB - A case of a 62 year old male with acute type De Bakey I aortic dissection is presented. The ascending aorta, aortic valve and total aortic arch replacement were performed. Additionally the proximal descending aorta was replaced using elephant trunk technique, to close the entrance of the dissection. There was no patent false lumen in distal descending aorta in postoperative findings. PMID- 12731386 TI - [Diagnostic problems in osteosarcoma first diagnosed as myositis ossificans]. AB - The aim of the study is presentation of problems in osteosarcoma diagnosis. In 3 cases possibilities of mistakes especially in differential diagnosis with myositis ossificans are presented. PMID- 12731387 TI - [Mesothelioma as delayed consequence of asbestos pollution]. AB - Mesothelioma is a relatively rare cancer, who's course is dynamic and leads to cahexia and patient death. The relationship has been proven between its incidence and exposition to asbestos. We presented six cases of mesothelioma in patients without professional risk, which were hospitalized in the internal sections of Pulmonological wards. Unfortunately, administered treatment did not bring expected results. PMID- 12731389 TI - [Issues of medical ethics]. PMID- 12731388 TI - [Jan Mikulicz-Radecki: 150th birth anniversary]. PMID- 12731391 TI - [The role of nuclear cell matrix in spatial organization of nuclear processes]. PMID- 12731390 TI - [Segregation of genetic material in prokaryotic cells]. PMID- 12731392 TI - [Import of protein into mitochondria]. PMID- 12731393 TI - [Lipoxygenase superfamily--structure and function in metabolism]. PMID- 12731394 TI - [Structure and role of the tau protein]. PMID- 12731396 TI - [Biological activity of galanin and its significance in physiologic and pathologic processes]. PMID- 12731395 TI - [Prolactin (PRL)--multifunctional, hypophyseal peptide hormone]. PMID- 12731397 TI - [Determination of hormones in blood plasma by HPLC techniques]. PMID- 12731398 TI - [Requirements and quality of health promotion in the workplace--evaluation study]. AB - This paper presents the results of an attempt to assess the quality of voluntary activities aimed at improving the workers' health in Polish large and medium sized enterprises in view of the health promotion requirements. Based on the data collected via a questionnaire mailed to enterprises selected at random in 1998 (147 enterprises), 2000 (755 enterprises) and 2001 (215 enterprises) the determinants of the health promotion quality are discussed. A long standing tradition to protect the workers' health was reported by 50% of enterprises as the major reason for undertaking health promotion activities. Only 25% of enterprises described their activities as health promotion understood as a tool for supporting their market strategy through well coordinated voluntary actions aimed at improving health and well-being of their employees, however, 85% of respondents reported the implementation of various health conducive programs. The most frequent failures of health-oriented activities to be of a required standard are: (a) regarding these activities as those of little importance compared with the enterprise priorities; (b) lack of a special section responsible for health promotion issues in an enterprise; (c) ignoring the employees' right to participate in planning and evaluating health promotion programs, which means that they are perceived as passive service recipients or passive performers of tasks assigned to them; (d) lack of cohesive approach towards health problems and strong belief that all the goals can be achieved through restrictions or education without any support; and (e) reluctance of enterprises to evaluate the organization, implementation and outcomes of health conductive activities. The major obstacle to health promotion is lack of money (reported by 60% of enterprises) and lack of statutory obligations (reported by more than 50% of enterprises). The former is usually reported by enterprises that are not active in this field, and the latter is an obstacle to enterprises that have invested in workplace promotion programs. PMID- 12731400 TI - [Development of carpal tunnel syndrome after being certified and an occupational disease]. AB - The authors present the results of the study carried out in South Moravia, Czech Republic, in a group of 140 patients (76 women and 64 men) with occupational carpal tunnel syndrome (CTS). The time-lag between the occupational disease certification and the investigation ranged from 2 to 14 years. The mean age of persons at the diagnosis was 44.5 +/- 7.4. years, and the duration of hazardous occupational exposure was 19.8 +/- 9.8 years. Repetitive and excessive strain of upper extremities was observed in 59% of patients and locally transmitted vibrations to the hands or vibration combined with repetitive and excessive strain of upper extremities in 41% of patients. Following the CTS certification as an occupational diseases, 98% of patients were removed from exposure to etiologic factors. During a follow-up examination, 19% of patients evaluated their health condition as better or recovered, deterioration was reported by 28%, and unchanged condition by 63% of subjects. Surgery was performed on 61% of patients. The comparison of the examination outcome at CTS certification as an occupational disease with the follow-up examination indicated significant improvement of distal motor latency, but the complete recovery of the median nerve conduction was found only in 13% of patients. PMID- 12731399 TI - [Sick leave among workers employed in restructured enterprise]. AB - Ownership and restructuring transformations that are taking now place in Poland, as well as the situation on the labor market have their impact on the indicators, which illustrate the workers' health situation, including temporary work disability preceding the issue of the certification granting the disability pension. The aim of this analysis was to identify the changes in the extent and causes of sickness absence among workers after restructuring. The study was carried out in one of the largest transport industry enterprises during the years of its restructuring (1984-1994), covering 8588 workers, and after its restructuring (1997-1999), covering 2702 workers. Following the restructuring, the enterprise's staff was rejuvenated so that the number of workers aged over 50 years decreased by almost fifty percent. The analysis was based on the sickness absence rate calculated as the ratio between the number of days of work disability in a given period of time and the number of person-days in the same period. In 1997-1999, a 33% decrease in sickness absence among women and a 25% decrease among men were observed in the study enterprise as compared with the period of 1989-1994. However, the enhanced absence was also found due to the following diseases: mental disorders (a threefold increase in men); diseases of the musculoskeletal system (by 54% in men and by 43% in women); endocrine, nutritional and metabolic diseases and immunity disorders (a threefold increase in women). Following the restructuring, considerable changes in the sickness absence structure, by causes of diseases, were revealed. A substantial decrease in the share of male and female absence due to diseases of the respiratory and circulatory systems and almost threefold decrease in complications of pregnancy, childbirth and the puerperium in women were noted. In addition, over twofold increase in male and female sickness absence due to diseases of the musculoskeletal system, and lower but significant increase due to mental pathologies (from 3 to 12% in males and from 4 to 7% in women) were recorded. The changes observed in sickness absence result mainly from the changes in the magnitude and structure of employment in the restructured enterprise and related organizational modifications, which arise stressogenic situations among workers, more intensified at risk of unemployment. PMID- 12731401 TI - [Protective effect of melatonin and vitamin E against prooxidative action of iron ions and static magnetic field]. AB - The purpose of this study was to examine the effect of melatonin and vitamin E (trolox) on the level of lipid peroxidation in rat blood lymphocytes after in vitro (3 h) exposure to iron ions and/or 7mT static magnetic field (SMF). The lipid peroxidation process was chosen as a marker of free radical mechanism of SMF in cells. The cells were supplemented with (0.5 mM) melatonin or (0.1 mM) vitamin E (trolox) in preincubation. During SMF exposure in Helmholtz coils some samples were treated with ferrous chloride (10 mg/ml or 20 mg/ml), while the rest served as controls. There is a significant increase in the amount of lipid peroxidation end-products (4-HNE + MDA) in rat lymphocytes after simultaneous exposure to 7 mT SMF and iron ions (versus control samples and those exposed to SMF alone). Instead, when the cells were treated with melatonin or trolox and then exposed to iron ions and 7 mT SMF, the level of lipid peroxidation was significantly reduced. The results also indicated that melatonin is less effective than vitamin E (trolox) in inhibiting lipid peroxidation under the experimental conditions used. PMID- 12731402 TI - [Influence of pulsating magnetic field used in magnet therapy and magnet stimulation on cortisol secretion in human]. AB - The aim of our study was to test the influence of magnetic fields during magnetotherapy and magnetostimulation over a longer period of time (like in physiotherapy) on cortisol secretion in humans. The study population was divided into two groups: magnetotherapy group (16 men) and magnetostimulation group (10 men). Magnetotherapy in the form of magnetic field induction (2.9 microT; frequency--40 Hz; square wave; bipolar; Magnetronic MF--10 apparatus) was applied for 20 min to the lumbar area in patients with chronic low back pain. Magnetostimulation (Viofor JPS system; M2P2 program; induction--25-80 microT; frequency--200 Hz, complex saw-like shape with a plateau halfway the height of the wave; bipolar) was applied every day for 12 min in patients with the same health problem. In both groups, the procedures were repeated 15 times (about 10:00 a.m.) with weekend breaks. Serum samples were collected at 6:00, 12:00, 16:00 and 24:00 and estimated by the micromethod of chemiluminescence (DPC Poland; Cat. No. LKC01). The circadian profile of cortisol was determined prior to the application, a day and a month after application. The data were analyzed statistically, using paired and unpaired Student's test. Magnetotherapy affects the cortisol secretion in the circadian profile by decreasing its level at 16:00 a day after 15 applications, whereas magnetostimulation by increasing its level at 12:00 a month after 15 applications, which may suggest its long-term effect on hypothalamic-pituitary axis. The comparison of the results indicated that a day after magnetotherapy and magnetostimulation, the circadian curves of cortisol secretion differed significantly by about 100%. All hormone oscillations did not exceed the physiological norms of the circadian cortisol level, not reaching the level so high as in an intense stress. This suggests rather their controlling effect on the cortisol level than their significant stressogenic nature. PMID- 12731403 TI - [Working condition of pregnant women. Departures from regulation on occupations especially noxious or hazardous to women]. AB - To define the range and kind of discrepancies between regulations and actual state of working conditions and occupations performed by women during pregnancy, a survey was carried out in the population of 2649 women, a representative group of occupationally involved pregnant women in the region of Lodz. The questionnaire analysis provided information on the kind and arduousness of work performed, working hours and conditions in the work environment. Having verified the data obtained from 10% of women, a very high consistency between working conditions described by respondents and evaluated by experts was noted. Its was found that almost 60% of women under study indicated that their working conditions depart from those defined as admissible for women during pregnancy. Of the women who worked only during the first month of pregnancy, 75% of them were employed at workposts, which did not meet the requirements, and nearly half of women who worked during the whole period of pregnancy were employed in inappropriate conditions. The noxiousness most frequently reported by respondents applied to body posture: bending and position with rotated spine (about 40%). This was followed by lifting and carrying various objects (about 30%), prolonged working hours (about 20%), shift work, including night shifts (about 12%), work at conveyor belt and forced work pace (about 20%). Noise that makes it difficult to communicate, local vibration and work with computers over 4 hours daily was reported by 30% and 15% of respondents, respectively. The most common solution to limit the potential effect of hazardous and noxious working conditions on pregnant women was to remove them from work by issuing a sick leave certificates, while the modification of working conditions or shifting them to another post applied only to 14% of women. The results of the study showed that working conditions of pregnant women differed from those described by the State Labor Inspectorate. The discrepancy can be explained by different method of data collection. PMID- 12731404 TI - [Relative risk of pneumoconiosis in welders in metallurgy]. AB - The values of pneumoconiosis risk in welders calculated against the dust doses that induce this pathology do not show linear relationship. In the group of electrical welders, relative risk (RR) was found statistically significant at the doses of 251-500 g; 1001-1500 g and 5501-6000 g. In the group of semi-automatic welders, similar results were observed. Odd ratio, calculated at workposts of semi-automatic welding with CO2 shield, showed that, depending on the size of a daily dose of dust, statistically significant RR was found at the doses of 1.6 2.0; 2.1-2.5; and 2.6-3.0 mg/kg/day. In the group of electric welders, statistically significant RR was observed at the doses of 2.1-2.5; 6.1-6.5; and 9.1-9.5 mg/kg/day. This may suggest that welding dust at workposts of semi automatic welding is more aggressive. The division of welders by their dates of birth showed that in the group of welders born by 1945, the mean age at which they developed pneumoconiosis was almost 50 +/- 0.4 years, and the mean duration of occupational exposure was 25 +/- 0.3 years. In the group of welders born after 1945, these values were 36 +/- 0.6 years and 12.8 +/- 0.2 years, respectively. PMID- 12731405 TI - [Financing of regional occupational health service centers: structure and financial criteria in years 2000-2001]. AB - The rational planning and financing of occupational health services at the national level have to be based on an appropriate system of information about individual units and their financial status that could illustrate their financial administration. This is required not only in view of the internal needs of public money management, but also in view of the national health accounts. The major task in this regard is to assess the level and structure of financing to individual units and to check the soundness of criteria used in the process of supplying financial means. The results of such an analysis can be a valuable source of information for planning carried out also by the institutions which provide funds to cover the cost of tasks performed by individual units. The aim of the project implemented by the Nofer Institute of Occupational Medicine was to collect, process and analyze data on the level and structure of financing of provincial occupational medicine centers. In this paper, the objectives, methodology and analytical tools are discussed. The results and structural data on the level and structure of financing of regional occupational health services centers covering a two-year period are presented. At the same time, the criteria for allocating funds were identified, which made it possible to evaluate the situation and to propose new solutions. PMID- 12731406 TI - [Bronchial asthma in rural environment]. AB - This paper reviews basic problems concerning epidemiology and pathogenesis of bronchial asthma in the rural environment. Possible causes of the decreased prevalence of atopy and allergic diseases are analyzed. Etiology, pathogenesis and clinical findings of asthma-like syndromes, a major problem of differential diagnosis, as well as the principles of occupational asthma diagnostics in farmers are also presented. PMID- 12731407 TI - [Health effects of occupational exposure among shoe workers. A review]. AB - World-wide epidemiological studies provide evidence that the employment in the shoe production and repair plants is associated with an enhanced risk for cancer (primarily nose and nasal sinuses). According to the majority of authors, it is induced by exposure to leather dust. It is also known that, leather dust particles contain numerous chemicals acquired during the process of leather tanning and finishing (chromium salts, vegetable dye extracts, mineral oils). Some of these compounds exert carcinogenic effect. This paper provides a review of the results of epidemiological studies on health effects of exposure to harmful factors present mainly at the footwear production and repair. These results reveal an enhanced risk for cancer of nose or nasal sinuses induced by leather dust, as well as neoplasms of hematopoietic and lymphatic systems, resulting from exposure to solvents (mostly benzene). Among non-neoplasms, diseases of the musculoskeletal system associated with ergonomic factors, contact dermatics, chronic pulmonary diseases and damage of peripheral nerves in solvent exposed workers are diagnosed. PMID- 12731408 TI - [Harmful agents in municipal wastewater treatment plants]. AB - Chemical and biological hazards and health effects of exposure to some selected agents on the workers of municipal wastewater treatment plants are reported. This issue has not as yet been thoroughly analyzed in Poland. The workers of wastewater treatment plants are exposed to: heavy metals (Pb, Cu, Zn, Cd, Cr, Ni), polycyclic aromatic hydrocarbons (PAH), polychlorinated biphenyls (PCB), volatile organic matter, hydrogen sulfide, organic agents, including bacteria, viruses and fungi, organic-produced endotoxins and glucans. The most frequent disorders reported by the wastewater--treatment workers comprise respiratory and gastrointestinal infections, ocular and dermal irritations, headache and discomfort. The assessment of hygienic conditions at individual workplaces is difficult because of lack of hygiene standards for airborne microorganisms and endotoxins. Up to now, the exposure of workers employed in municipal wastewater treatment plants has not been assessed in Poland. The number of the affected workers is estimated at 11,000-19,000 people. PMID- 12731409 TI - [Factors modifying noise-induced hearing loss]. AB - This paper introduces the current data on the factors, which determine individual susceptibility to noise. Pathophysiology of occupational noise-induced hearing loss is nowadays a well-known issue. Both experimental and clinical data presented, show the variety of factors influencing the natural history of hearing impairment in workers employed in various branches of industry. It is necessary for an occupational medicine specialist to consider these factors in preventing hearing impairment as noise, according to epidemiological data, is the most common harmful factor in the work environment. PMID- 12731410 TI - [Lead effect on vascular endothelium]. AB - The results of numerous epidemiological and experimental studies show that environmental exposure to lead in humans, as well as small doses of this element given to experimental animals exert hypertensinogenic and athoregenic effect. The vascular endothelium, involved in the development of arterial hypertension and arteriosclerosis, is now regarded as the main target organ for the toxic effect of lead. This metal can influence endothelium in various ways. Functional disturbances, mainly in respect of synthesizing and modifying functions of intravascular coagulation processes, impairment of endothelium integrity and cytotoxic effect have been observed. The lead-induced vascular damage may involve the inhibition of the repair processes of endothelial cells damaged by various exo- and endogenous factors. In addition, lead can also affect, directly or indirectly, the vasoactive function of endothelium through the increased production of reactive oxygen species. This effect of lead results in modified nitric oxide or endothelin synthesis and/or their release. Lead also accelerates the proliferation of smooth muscle cells and disturbs the synthesis of prostacyclins. PMID- 12731411 TI - [Efficacy of the health promotion program "Prevention of Voice Disorder in Teachers and Students" implemented by the WOMP from Zielona Gora ]. PMID- 12731413 TI - [Epidemiologic and health economic significance of colorectal cancers in Germany]. PMID- 12731412 TI - [Carcinogenesis and hereditart colon cancers]. PMID- 12731414 TI - [Colorectal cancer in Germany. Means for prevention and early detection: implications for laiety and physicians]. PMID- 12731416 TI - [Conventional and virtual colonoscopy]. PMID- 12731415 TI - [Conventional and molecular screening (fecal tests)]. PMID- 12731417 TI - [Minimal invasive therapy of "early" tumors]. PMID- 12731418 TI - [Surgical therapy of colorectal carcinoma]. PMID- 12731419 TI - [Multimodal therapy in colon carcinoma]. PMID- 12731420 TI - [Parasite detection and symptoms of parasitic diseases. 1: Blood parasites]. AB - Blood parasites are malaria plasmodia, microfilaria species, trypanosomes (the causative agents of African sleeping sickness and South American Changas disease) and the causative agents of schistosomiasis of the bladder and the intestine. Their geographical distribution, incubation periods, signs and symptoms, microscopic and serological methods are described. In Germany around 1,000 tourists contract malaria every year, mostly travellers to Africa. Over 70% suffer from the life-threatening P. falciparum infection. Only a few days after the onset of this flu-like disease, complications may evolve. The best diagnostic method is the thin blood film. In case of a negative result this procedure must be repeated twice daily. The thick film requires experience. Rapid diagnostic tests can be helpful but are hampered by false negative results. Filaria loa loa may cause skin swellings, involvement of the eye and even the CNS; Wuchereria bancrofti can cause severe lymphedema. West African sleeping sickness (Trypanosoma gambiense) ends up in encephalitis, the East African form (T. rhodesiense) in a polyserositis. Schistosomiasis of the urinary bladder and the large intestine may cause severe diseases of the urinary tract or the liver. PMID- 12731422 TI - [Hepatitis of unknown origin with protracted liver failure in a 48-year-old patient with significantly increased pANCA titer]. AB - In spite of intense diagnostic testing, no cause for the chronically aggressive hepatitis of a 48-year old male patient was found. Evidence for an autoimmune process, however, could be derived from a high titer of pANCA. Only according to the revised criteria of the working group on autoimmune hepatitis, but not to the first version, it was possible to classify this as an autoimmune hepatitis. Despite of high-dose steroid treatment and accelerated preparation for liver transplantation the patient died of the complications of rapid liver failure. Thus, in case of unclear rapid progressive hepatitis, the revised criteria of autoimmune hepatitis should be reviewed early and with high priority and consequent high-dose steroid therapy and preparation for liver transplantation should be initiated. The prognostic impact of a high titer of pANCA in patients with autoimmune hepatitis remains to be established. PMID- 12731421 TI - [21-year-old patient with myocardial infarct, transient cerebral ischemia and thrombocytopenia]. AB - We report on a case of a young female soccer player who first experienced a transient ischemic attack at the age of 21 and then suffered an acute myocardial infarction accompanied by thrombocytopenia 6 months later. The underlying cause was antiphospholipid syndrome. This clinical picture is caused by a very heterogeneous group of antiphospholipid antibodies, which interfere with plasmatic coagulation at various sites and are able to activate thrombocytes, endothelial cells, and monocytes. This leads to a thrombophilic condition with clinical manifestation of recurrent venous thromboses, arterial embolisms, or premature birth or miscarriage. The main therapeutic options are thus anticoagulation, heparins, and aspirin. Steroids are recommended for cases of pronounced thrombocytopenias; if treatment fails, immunosuppressants are also advisable. The antiphospholipid syndrome should be kept in mind especially in cases of young patients with thromboembolic events since the presence of antiphospholipid antibodies can be detected in 10-20% of this patient group and effective therapy after diagnosis lowers the rate of recurrence. PMID- 12731425 TI - [Stenting ovation?]. PMID- 12731424 TI - [Reduction of cardiovascular events with lorsartan? The LIFE Study]. PMID- 12731423 TI - [Weight loss via drug therapy]. AB - Obesity and its associated diseases are an increasing challenge in medicine. A change in lifestyle is usually the first step with modifications in nutrition, physical activity and behavior. However, most of obese patients are not able to follow such a treatment regimen for a longer period of time. If they do not lose > 5% of initial weight within 3-6 months, pharmacological intervention should be taken into account. Orlistat, a gastro-intestinal lipase inhibitor, enhances fat excretion thereby reducing energy uptake and body fat. Studies up to 4 years document a net weight loss of 3-5 kg, all cardiovascular risk factors are reduced. Sibutramine, a serotonin- and noradrenalin reuptake inhibitor, promotes satiety and stimulates energy expenditure. Within one year a net weight reduction of 4-6 kg is achieved and morbidity as well as quality of life are improved. For both drugs no end-point outcomes are available so far. The anti-obesity drugs orlistat and sibutramine are useful tools for overweight and obese patients as an adjunct to lifestyle changes. Under the supervision of experienced physicians the combined treatment consisting of non-pharmacological and pharmacological methods reduces body weight in more than half of the patients and improves morbidity and quality of life. PMID- 12731426 TI - [Coronary stents]. AB - Since the introduction of coronary stents into clinical practice in the late 1980s, the number of stent implantations has increased so rapidly that stents are currently used in over 80 percent of all percutaneous coronary interventions. Although stent implantation was initially limited to large vessels with proximal and discrete lesions, improvements in stent design and implantation technique now allow their deployment in more complex lesions in smaller and diffusely diseased vessels. The overall acceptance of stents by interventional cardiologists can be attributed to favorable acute and longterm results compared to balloon angioplasty alone. Interventionalists have also been quick to embrace the smoother and larger lumen after stenting, in a shorter procedure time and with no additional risk, especially since the risk of stent thrombosis has been overcome by the introduction of dual antiplatelet therapy with Aspirin and Ticlopidine or Clopidogrel. Although restenosis and the need for reinterventions is lower after stenting compared to balloon angioplasty it still remains significant with about 15 percent of all patients returning for an other revascularization procedure. Meanwhile, a completely new generation of stents promises to eliminate the problem of restenosis. Drug-eluting stents, coated with antiproliferative substances have been successfully tested in small randomized trials. The restenosis rates at 6 and 12 months were extremely low ranging between zero and nine percent, with no clinical drawbacks so far. If these results hold up in longer follow up and in real life practice with more complex lesions stented the treatment of symptomatic coronary artery disease will change even more dramatically. PMID- 12731427 TI - [Stenting and stent-grafting of the aorta]. AB - The therapeutical approach to diseases of the descending thoracic and abdominal aorta has dramatically changed over the last decade due to new stent-graft-based endoluminal techniques. In particular elderly patients with a variety of diseases, such as ruptured and asymptomatic aneurysms, acute B-dissections, acute penetrating ulcers, mycotic aneurysms or traumatic aortic injuries will benefit from these minimally invasive alternatives, which can be performed under local anesthesia. As the durability of these devices is not yet proven the patients undergoing endoluminal aortic repair will need life-long clinical and imaging follow-up. PMID- 12731428 TI - [Stents in the treatment of supraaortic vessel stenosis]. AB - We describe and discuss our experience of more than 500 endovascular procedures for revascularisation of acute or chronic occlusion and stenosis of supraaortic vessels using stents in 171 cases. Whereas endovascular treatment of the innominate, subclavian and vertebral arteries are routinely used in atherosclerotic occlusive lesions, carotid stenting is currently being investigated as an alternative treatment to carotid endarterectomy and seems to offer a less invasive, less traumatic and less cost expensive alternative to achieve the goal to prevent stroke. In a subgroup of high surgical risk patients carotid artery stenting with simultaneous embolic protection of the cerebral blood flow by filter systems was even superior to the endarterectomy-treated patients in a prospective, randomized multi-center study (SAPPHIRE): at 30-day follow up the major event rate for the stented group of 156 patients was 5.8% versus 12.6 for the 151 endarterectomy-treated patients. PMID- 12731429 TI - [Stents in peripheral arterial occlusive diseases (PAOD)]. AB - Stent angioplasty has become a routine procedure in endovascular therapy. The indication must be evaluated in direct comparison with balloon angioplasty and vascular surgery. Technical and clinical success rates in various vascular regions must be considered as well as the costs induced by stent implantation. Stents for iliac artery disease are indicated after failed or complicated balloon angioplasty. If accepted indications for endovascular treatment are closely followed, we can expect a stent rate of approximately 40%. For the femoropopliteal segment stent angioplasty is only acceptable for very selected cases, mostly severe dissections; stenting of these vessels is not generally accepted. This review summarizes indications, technical and clinical success rates of stent angioplasty for peripheral arterial disease. PMID- 12731430 TI - [Venous stents]. AB - The continuous development of stents in the past 20 years has facilitated the widespread use of stents in the arterial and venous system. The idea of stents is tightly related to the name of Charles Dotter, the inventor of percutaneous transluminal angioplasty (PTA). Stents can be placed for various causes of obstruction, both benign and malignant, throughout the venous system either, primarily, in combination with fibrinolysis, or as adjunct to PTA or surgical intervention. Stent placement is a minimal invasive method associated with a low complication rate and a high clinical success mainly due to differences of the properties and of the pressure between veins and arteries. Special cases of veinous stents are hemodialysis shunts, where patency rates are not improved but critical situations can be managed, and transjugular intrahepatic porto-systemic shunts (TIPS), which have high rates of complication and restenosis requiring frequent reinterventions. PMID- 12731431 TI - [Stents in the tracheobronchial tree]. AB - Interventional procedures in bronchoscopy such as tissue ablation by laser, bronchusdilatation, stenting, endobronchial radiotherapy have a firm established position as treatment modalities for severe tracheobronchial obstruction. They allow quick and effective treatment of sometimes life threatening tracheal and bronchial obstructions. This article focuses on stents in the airways. A brief description of historic development is given. Currently available and frequently used stents are shown. There advantages and disadvantages are discussed. Indications for Stent-insertion in particular and for interventional endoscopy are summarized and the different techniques for insertion outlined. Based on the published literature results and complication rates are discussed. PMID- 12731432 TI - [Esophageal and intestinal stents]. AB - Implantation of expandable metal stents has evolved during the last decade as an established method for the palliation of tumourous obstruction within the gastrointestinal tract. Rigid plastic stents have initially been used for the treatment of dysphagia due to esophageal carcinoma. The development of different types of self expandable metal stents with differing physical characteristics has facilitated their use for the palliative treatment of not only esophageal but gastroduodenal and colonic obstruction as well. The hallmark of these devices is the thin diameter of the compressed stents which allows introducing them into the obstructed segment without the need of aggressive dilatation. Once in place their compression is released and they self expand to a final diameter of approximately 20 mm. Perforation thus occurs rarely and these stents may successfully be placed in over 90% of patients with complete relief of obstruction in over 80%. The most frequent complications are stent dislocation and stent obstruction due to food impaction or tumour overgrowth. PMID- 12731433 TI - [Biliary stents]. AB - Endoscopically implantable stents are today the mainstay for therapy of biliary stenoses. It is important to know if a benign or a malignant stenosis is the cause of the biliary obstruction. Generally benign stenoses are treated with plastic-stents whereas malignant stenoses are managed by implantation of a metallic stent. The main indications for plastic stents are postoperative strictures in the biliary tree for example, after biliary tract surgery or liver transplantation, primary sclerosing cholangitis and postoperative biliary leakage. Metallic stents are implanted in palliative circumstances like in stenosing cholangiocarcinoma or in situations where a hepatic metastasis exerts an extrinsic compression on the biliary tract with consecutive cholestasis. The materials used for manufacturing both stent types are biologically inert and thus biocompatible. A current poorly resolved problem is the occlusion of the stent lumen (by sludge, bacterial degradation products etc.) which occurs in both stent types with time. These problems lead to stentocclusion around three to six months after implantation and necessitate endoscopical re-interventions in order to overcome the occlusion. The patency rate for metallic stents is better than for plastic ones. The endoscopic stenttherapy is equivalent to surgical therapy (Intestinal bypass-procedures). PMID- 12731434 TI - [Stents in urology]. AB - Ureteral stents are employed in the upper and urethral stents in the lower urinary tract for restitution or maintenance of urinary drainage. Placement of ureteral stents is indicated as an adjuvant measure prior to extracorporal disintegration (ESWL) of large kidney stones to insure urinary drainage and enhance expulsion of fragments and disintegrate. Also, obstruction by very small urinary tract stones that are not treatable by ESWL because they cannot be localized can be relieved by placement of a double-J-stent with immediate elimination of colic. If the cause of urinary tract obstruction is external ureteral compression (retroperitoneal mass), placement of a special tumor stent is one possibility. This, however, has the danger of becoming reobstructed with detritus and blockage of the drainage holes in the stent. In these cases the essential drainage along the stent is blocked by the mass. Therefore, a percutaneous nephrostomy providing direct drainage is easier to control and preferable. Obstructive pyelonephritis is an absolute indication for drainage of the upper urinary tract with a double-J-stent, or even better by percutaneous nephrostomy. If pyeloureteral or ureteral stenoses of the upper urinary tract are opened endoscopically, then the double-J-stent serves to maintain and insure drainage until the new lumen is reepithelialized. In patients with prostatic hyperplasia who no longer respond to medical treatment and who are not candidates for more invasive surgical treatment, a stent can be placed in the prostatic urethra under local anesthesia as a last resort. This procedure is seldom used but, in view of the satisfactory long-term results, it provides a true alternative to bladder drainage by transurethral catheter or percutaneous cystostomy. The same stents may be used in the bulbar urethra to reduce restricture rates following endoscopic treatment of strictures. PMID- 12731435 TI - [Localized prostate cancer]. PMID- 12731436 TI - [Hypogastric artery embolization as a palliative treatment for bleeding secondary to intractable bladder or prostate disease]. AB - OBJECTIVES: Bleeding secondary to severe vesicoprostatic pathology (mainly neoplasic disease and radiation cystitis) may be a serious clinical management problem due to its morbidity and associated increased resources demand in the form of admissions, transfusions and other measures. We review a series of patients embolized for this purpose, its efficacy, tolerability and adverse events. METHODS: We review 8 patients who underwent hypogastric arteries embolization between July 1998 and December 2001, analyzing indications, efficacy and duration, tolerability, and consequences. Right femoral artery access was undertaken in all except one case that needed bilateral femoral accesses. Embolization was achieved by means of coils and particles. All procedures were performed under local anaesthesia. RESULTS: 9 procedures were performed in 8 patients. 3 patients presented with hematuria due to radiation cystitis, 3 from prostatic carcinoma, and 2 with urethrorragy from urethral relapses after cystectomy. 7/9 embolizations were effective achieving immediate complete or almost complete bleeding control; in one case control was partial; another one had limited or no effect. Selective embolization of distal arteries was performed in 3 cases; all the remainders underwent direct hypogastric trunk embolization sparing the superior gluteal artery. Effect lasted between 1 and 31 months. 4 patients died, 3 of them without haematuria, 1 because of an intercurrent disease, and the others from disease progression. 2 patients underwent posterior surgery, one due to recurrent haematuria, and the other, a case of urethral tumour, due to partial failure; partial cystectomy and urethrectomy were performed respectively. 2 patients needed administration of morphic derivatives after embolization, all the others were managed with magnesium metamizol. Only one patient referred mild transitory gluteal claudication. CONCLUSIONS: Percutaneous arterial embolization is an effective instrument to treat patients with haematuria or urethrorragy and severe lower urinary tract pathology in whom curative treatments are not applicable due to their general status, life expectancy, or tumor status. PMID- 12731437 TI - [Ureteral lesions in gynecologic surgery]. AB - OBJECTIVES: Ureteral lesions during gynaecologic surgery are a serious problem, affecting morbidity even when they are diagnosed postoperatively. METHODS: 742 gynaecological surgical procedures performed between 1994 and 2000 at the Manuel Fajardo University Hospital where evaluated. Eight operations were complicated with ureteral lesions (1.07%); one patient suffered double lesions, for a total of 9 lesions. RESULTS: 9 ureteral lesions were treated in 8 patients, 7 of them were diagnosed in the postoperative period; 5 bladder reimplantations with submucosal bladder tunnel and 2 ureteral suture repairs (end to end) were performed. Delayed lesions were 2 ureteral-vaginal fistulae treated with nephrectomy and ureteroneocystostomy into a Boary's flap respectively. In both cases diagnosis was established by combination of ultrasound, intravenous pyelogram, cystoscopy, and retrograde catheterization. In all seven patients urinary tract integrity was demonstrated postoperatively. CONCLUSIONS: Lower urinary tract integrity may be damaged during gynaecologic surgery, being bladder ureteral reimplantation with submucosal tunnel the most frequently used repairing procedure. Operations for benign diseases had fewer lesions (0.47%) than malignant (4.8%). PMID- 12731438 TI - [Quality control plan for a primary bladder neoplastic process]. AB - OBJECTIVES: Delays in the diagnosis and treatment of bladder cancer may change survival rates (6). We present the application of a plan for quality control in the management of primary bladder cancer, studying delays globally, and for individual indicators, as well as the impact of implemented improvement measures. METHODS: We performed a retrospective chart review to detect problems appeared through the whole clinical management process in a population of patients undergoing surgery for primary bladder cancer in the Hospital Costa del Sol during 2001. Causes were studied and corrective measures established, application of which has been evaluated during the first semester of 2002. Study indicators encompass from delay time for first visit to delay time for radical surgical treatment. RESULTS: Excessive average global delays for radical treatment of bladder tumor (208.3 days), and variability in the time to transurethral resection (one to 73.42 days) and to postoperative revision (54.4 days), were the most significant results. Evaluation of implemented measures shows a descent in global delays (134.2 days) and time to postoperative revision variability (32.4 days). CONCLUSIONS: Application of quality control procedures for clinical management is essential in daily clinical practice, for systematic, continuous, and rigorous evaluation will allow us to offer patients a product that satisfies their personal and social expectations. PMID- 12731439 TI - [Utility of computerized tomography in determining the extent of infiltrating bladder tumors: our experience]. AB - OBJECTIVES: To evaluate the reliability of bladder transitional cell carcinoma (TCC) staging, comparing findings on computerized tomography (CT) with pathologic results after radical cystectomy. METHODS: We retrospectively review 115 consecutive patients with bladder TCC undergoing radical cystectomy. Preoperative CT findings were compared with pathology results obtained after cystectomy and lymphadenectomy. RESULTS: We found that as a whole CT showed a tendency to local overstaging of 27.8% for tumours infiltrating bladder wall only, and understaging of 36.5% Regarding lymph nodes involvement, CT overstaged 4.6% of patients and understaged 23.8%. CT was capable to detect only 7.15% pN positive cases. CT was unable to detect the only patient with intra-abdominal metastatic involvement. CONCLUSIONS: Abdomino-pelvic CT has real limitations to detect extra vesical extension and lymph node metastasis in patients with bladder TCC. PMID- 12731440 TI - [Use of a urethral plate in surgery for incurving hypospadias]. AB - OBJECTIVE: To describe a new procedure for surgical correction of primary hypospadias with chordee. METHODS: The design of the urethral plate divide it in two flaps. Chordee must be resected in order to straighten the penis. Ventral side of neourethra is made with preputial skin or Mathieu procedure. This method has been used in 10 pediatric patients diagnosed of primary hypospadias with chordee. RESULTS: Medium follow-up was 6 months. Only 2 patients developed urethro-cutaneous fistulae and required a new surgical procedure. No reports of meatal stenosis, urethral dilatation or residual curvature were made. 8 patients have excellent cosmetic results and 2 cases were classified as acceptable. CONCLUSIONS: This procedure is a new surgical method for correction of primary hypospadias with chordee. PMID- 12731441 TI - [Renal transplantation in donors over 60 years of age]. AB - OBJECTIVES: To evaluate the impact of receptor's advanced age on kidney transplant outcomes. METHODS: We reviewed all transplants performed between January 1990 and December 1999. Among 570 patients receiving grafts, 115 patients were 60 years or older at the time of transplantation. We compared this group with receptors younger than 60 years. We studied possible prognostic variables and compared patient and graft outcomes. RESULTS: Mean age 63.81 (typical deviation (TD): 2.96). Mean follow-up time for elderly receptors was 41.6 months (TD: 26.58). 55.7% patients were males (p: 0.4). The most frequent cause for end stage renal disease was unknown etiology in group 1 and glomerular in the younger group (p: 0.01). 42% patients older than 60 years presented initial graft dysfunction, in comparison to 28.1% among younger than 60 (p: 0.006). Three-year graft survival was 90.42% for receptors 60 years old or older compared to 88.72% for group 2, without significant differences (p: 0.5). The most frequent graft loss etiology was patient death. (67.7%). (p = 0.005). Patient survival was 81.01% in group 1 and 95.25% in group 2, being differences significant (p < 0.001). CONCLUSIONS: Renal grafts in receptors over the age of 60 years show a greater incidence of delayed graft function, although it doesn't seem to influence final graft survival. The most frequent cause for graft loss is receptor's death. Receptor's age does not represent a contraindication for transplant. PMID- 12731442 TI - [Erectile dysfunction in those under 40. Etiological and contributing factors]. AB - OBJECTIVES: Although not very frequent, erectile dysfunction (ED) in patients younger than 40 years represents a disease to be kept in mind in our sanitary area. We propose a study of the various risk factors identified in this population group. METHODS: We studied a total of 736 patients with diagnosis of ED following our protocol, using as inclusion criteria being between 19 and 40 years of age. In this group, we have studied the incidence of various risk factors associated with the disease both globally and dividing the group into age subgroups also. RESULTS: Overall, we have diagnosed 64 patients with ED under the age of 40 years (8.7%). 25 of these patients were classified as psychogenic origin dysfunctions (39%), 26 organic (40.7%), and 13 of mixed origin (20.3%). Age subgroups analysis shows: between 19 and 25 years: 7 psychogenic, 2 mixed, and 5 organic; between 26 and 30: 1 psychogenic, 2 mixed, and 3 organic; between 31 and 35: 7 psychogenic, 3 mixed, and 8 organic; between 36 and 40: 11 psychogenic, 6 mixed, and 9 organic. We identified as risk factors, in order of prevalence: psychogenic (47%), diabetes (14.6%), cardiovascular (9.7%), toxic alcohol, tobacco, and drugs--(12.1%), neurogenic (4.8%), and others (7.3%). CONCLUSIONS: ED under the age of 40 years causes a growing demand of medical care day by day, mainly due to the increasing prevalence of its etiologic factors. As other publications have shown psychogenic factors are the most frequently associated with the disease in younger patients globally, although when dividing by age groups we found different results than other publications in terms of association with various risk factors. PMID- 12731443 TI - [Calcified uterine myoma. Presentation of 3 cases]. AB - OBJECTIVES: To report three cases of uterine myoma as cause of pelvic calcification and to establish differential diagnosis with bladder stones. METHODS/RESULTS: Three asymptomatic female patients, ages between 54 years (one patient) and more than 70 years (two patients), were fortuitously found to have calcified uterine myomas by radiological studies made for suspicion of other pathologies. CONCLUSIONS: Discovering of calcified uterine myomas is not frequent. Diagnosis is made by radiological studies in postmenopausal female patients. The significance from the urological point of view is to do differential diagnosis with bladder stones. PMID- 12731444 TI - [Primary ureteral leiomyosarcoma: a rare cause of obstructive uropathy]. AB - OBJECTIVES: Transitional cell carcinoma is the most frequent primary malignant neoplasia obstructive ureteral pathology. Nevertheless, obstructive primary ureteral leiomyosarcoma (stromal tumour) is an unusual disease, diagnosed by histopathological examination of nephroureterectomy surgical specimens. METHODS/RESULTS: We report a case of ureteral obstructive uropathy in a 38-year old woman, with non functional left kidney. Intravenous and retrograde pyelography revealed pyelocalyceal and ureteral dilatation and a large polypoid intraluminal lobulated mass in the pelvic ureter. Pathologic and immunohistochemical techniques showed a nonepithelial, stromal malignant tumour with appearance of smooth muscle cell differentiation: leiomyosarcoma. CONCLUSIONS: Primary leiomyosarcoma is an unusual ureteral obstructive etiology and should be considered in this condition. Correct preoperative diagnosis is rarely made and this tumour is diagnosed by pathological examination. Abdomino pelvic computerized tomography scan and ultrasound should be performed before diagnosing a lesion with these features as a primitive ureteral tumour. PMID- 12731445 TI - [Bifid ureter with a blind branch as cause of unspecified abdominal pain. Report of a new case and review of the literature]. AB - OBJECTIVE: To report a case of bifid left ureter with a blind-ending branch in a 35 year-old female patient who complained of poorly defined abdominal pain. METHODS: Clinical features, radiologic findings, differential diagnosis and therapeutic approach are discussed and the literature reviewed. RESULTS: Bifid ureter with a blind branch is not an uncommon anomaly as generally believed. There are at least a hundred and seventy-five cases reported in the literature reviewed. CONCLUSIONS: Diagnosis is most commonly made by intravenous urography (IVU) with oblique views showing the blind-ending segment filled by retrograde uretero-ureteral reflux. However blind segment does not always fill on excretory urography and retrograde pyelography is required for diagnosis (an ectopic kidney at the distal end of the blind-ending branch should be easily seen on CT scan or ultrasonography). A voiding cystourethrography (VCUG) may be needed to demonstrate an ureteral branch with coexistent vesicoureteral reflux. Treatment is initially conservative although complications or severe symptoms require surgical excision of the blind branch together with antireflux reimplantation of the normal ureter when vesicoureteral reflux is present. PMID- 12731446 TI - [Tuberculous epididymitis caused by Mycobacterium bovis]. AB - OBJECTIVES: To focus on the need of including tuberculosis among differential diagnoses of any epidymo-testicular mass, especially if its evolution is torpid. METHODS/RESULTS: A 73-year-old man who presented with scrotum abscess underwent surgical drainage and antibiotic treatment, but suppuration relapsed through cutaneous fistulae. A epipidymectomy was then performed, which demonstrated tuberculous granulomas. Torax Rx showed a cystic apical pulmonary wound which was treated with 3 antituberculostatics for 12 months. Sputum culture was positive for Micobacterium Bovis. CONCLUSIONS: Aspirative punction under sonographic control is a valuable technique to avoid mutilating surgeries and to permit an almost always effective treatment, before the appearance of permanent lesions which lead to sterility. PMID- 12731447 TI - [Neuroendocrine renal carcinoma. Presentation of a case]. AB - OBJECTIVES: To report a new case of neuroendocrine renal cell carcinoma. METHODS: We report the case of a 76-year-old woman with neuroendocrine renal cell carcinoma who underwent radical nephrectomy without any further adjuvant treatment. We performed a bibliographic review about this rare renal neoplasia of which there are less than 20 published case reports. RESULTS: Patient is asymptomatic four years after surgery, although she has local recurrence. CONCLUSIONS: Small cell renal cell carcinoma is a very rare neoplasia, affecting people over the age of 60 years, large in size, and metastatic at diagnosis. It has bad prognosis, with short survival times. The most adequate treatment has not been determined due to the scarcity of published cases; the combination of surgery and chemotherapy is the most frequently used. PMID- 12731449 TI - [High-flow priapism. Echographic diagnosis and treatment using selective embolization of the cavernous artery]. AB - OBJECTIVES: To report a case of high flow priapism and to review its diagnostic methodology and treatment options. METHODS: A 25-year-old patient consulted with prolonged erection after penile trauma. Colour Doppler ultrasound and angiogram were performed to confirm diagnosis. Cavernous artery selective embolization with reabsorbable material was performed. RESULTS: The case was resolved, and erectile function recovered. CONCLUSIONS: Colour Doppler ultrasound allows to diagnose high flow priapism. Angiography, in addition to diagnosis confirmation, allows performing selective cavernous artery embolization, which is currently the treatment of choice because it resolves the clinical picture preserving erectile function. PMID- 12731448 TI - [Extrinsic ureteral obstruction secondary to inflammatory gynecologic pathology]. AB - OBJECTIVE: To present two cases of extrinsic ureteral obstruction secondary to gynaecological inflammatory diseases, and to perform a bibliographic review. METHODS: We present two cases of obstructive uropathy with uretero-hydronephrosis secondary to tubo-ovarian abscesses diagnosed at our department. Clinical features at presentation, diagnostic tests, and preoperative management are reported. RESULTS: We report the clinical resolution of each case and perform a review about this pathology. CONCLUSIONS: Extrinsic ureteral obstruction is a frequent problem in urology. Inflammatory gynaecologic disease should be included among possible causes at the time of differential diagnosis. A methodical diagnosis process allows defining the exact location of the obstruction, diagnostic possibilities, and most adequate treatment plan for each case. Although association of gynaecological pathology and urinary tract obstruction is well known, there are not many bibliographic references in the national and international literature. PMID- 12731450 TI - [Nanobacteria and urinary system lithiasis]. AB - The presence of microorganisms in certain renal calculi having been demonstrated; currently, there is a debate about the possibility of certain germs of extremely small size, Nanobacteria, which can have an important role in the general theory of lithogenesis. PMID- 12731451 TI - Small cell carcinoma of the bladder: a case report and a literature review. AB - OBJECTIVE: To report a case of small cell carcinoma of the bladder. To analyse Literature data concerning disease stage, treatment and prognosis of small cell carcinoma of the bladder. METHODS/RESULTS: We report a case of small cell carcinoma of the bladder with hepatic and skeletal metastases who underwent radical cystectomy and systemic chemotherapy. The patient died 9 months after surgery because of chemotherapy complications and in disease progression. The Authors reviewed and analysed Literature data concerning 139 cases of bladder small cell carcinoma with available follow-up. CONCLUSIONS: The extreme variability of treatment options, the small number of cases and stratification by stages make it very difficult to deduce therapeutic recommendations. The management that seems to give a better survival is the combination of radical cystectomy (or radiotherapy) and chemotherapy. PMID- 12731452 TI - Transforming growth factor-beta 1 promotes contraction of collagen gel by cardiac fibroblasts through their differentiation into myofibroblasts. AB - Myofibroblasts and transforming growth factor-beta 1 (TGF-beta 1) are key elements of cardiac tissue fibrosis development. The aim of this study was to determine whether the ability of TGF-beta 1 to affect the contractile activity of cardiac fibroblasts depends on their differentiation into myofibroblasts. Cardiac fibroblasts (from male adult Wistar rats) from passage 2 were therefore cultured to confluency and incubated on a hydrated collagen gel, both with and without TGF beta 1 (0, 20, 40, 100, 200, 400 or 600 pmol/l), for 1, 2 and 3 days in a Dulbecco's Modified Eagle's Medium (DMEM) without fetal bovine serum (FBS). Growing cultures of cardiac fibroblasts were obtained by incubating second passage fibroblasts in DMEM with 10% FBS with or without TGF-beta 1 (0 to 600 pmol/l) for 6 days. These fibroblasts were then further incubated on the collagen gel for 1, 2 and 3 days in DMEM without FBS. TGF-beta 1 dose-dependently increased the contraction of collagen gel mediated by cardiac fibroblasts, either added directly to the gel or after growing of the cardiac fibroblasts in the presence of TGF-beta 1 for 6 days, reaching a maximal effect at 100 pmol/l TGF beta 1. In both culturing conditions, TGF-beta 1 also stimulated the [3H] thymidine incorporation and the total protein content in the cardiac fibroblasts in the collagen gel lattice. TGF-beta 1 dose-dependently induced an increase in alpha-smooth muscle actin, a marker of myofibroblasts, in both culturing conditions. The TGF-beta 1-induced reduction of area of the collagen gel was negatively correlated to the TGF-beta 1-evoked appearance of alpha-smooth muscle actin in the collagen gel matrix. TGF-beta 1 increased the contractile activity of adult rat cardiac fibroblasts and their ability to differentiate into myofibroblasts. Because contractile activity was correlated with differentiation, the influence of TGF-beta 1 on cardiac fibroblast-induced collagen gel contraction may depend on the promotion of myofibroblast differentiation. PMID- 12731453 TI - Anticonvulsant action of celecoxib (alone and in combination with sub-threshold dose of phenytoin) in electroshock induced convulsion. AB - Celecoxib is a selective cyclooxygenase-2 (COX-2) inhibitor, COX-2 has been shown to be upregulated by convulsive nerve activity. Various earlier studies have given conflicting reports on the effect of COX inhibitors on seizures. This study investigates the effect of pretreatment with celecoxib alone, or in combination with phenytoin, on electroshock-induced convulsions. Both percentage protection (i.e., the percentage of animals not showing Tonic Hind Limb Extension [THLE] when a fixed dose of current is administered) and CC50 (i.e., the threshold current inducing THLE in 50%) was determined using a technoconvulsometer. Celecoxib and phenytoin were administered 1 and 2 h, respectively, prior to the experiments. When administered alone, celecoxib showed an increase in percentage protection at increasing doses, with maximum percentage protection (66.6%) occurring at a 30 mg/kg-1 dose. The ED25 value of celecoxib was calculated to be 8.03 mg/kg-1. The CC50 values for the treatment groups were significantly increased compared with the control group (CC50 values for control, celecoxib 10 mg/kg-1, celecoxib 20 mg/kg-1 and celecoxib 30 mg/kg-1, respectively, were 36.3, 49.12, 100.3 and 125.02 mA). An increase in percentage protection was noted when celecoxib 8.03 mg/kg-1 was coadministered with phenytoin 6 mg/kg-1 (66.6% with the combination vs. 16.6% when administered individually). A significant increase was noted in the CC50 value in a combination regimen (CC50 = 79.06) compared with either drug administered alone (CC50 = 49.12 with celecoxib 8.03 mg/kg-1 and 62.06 with phenytoin 6 mg/kg-1). This study may stimulate further interest in the role of COX-2 inhibitors in the modulation of seizure activity. PMID- 12731454 TI - Involvement of nitric oxide in the nociception of kyotorphin, TYR-CAV and MIF-s analogues in the rat spinal cord. AB - The spinal analgesic effects of Kyotorphin (Kyo) and Melanocyte-inhibiting factor (MIF-l) were studied during acute pain in rats chronically implanted with intrathecal (i.t.) cannulas. Kyo (5 micrograms), t-Cav (5 micrograms), Tyr-Cav (5 micrograms), L-NAME (1500 micrograms), MIF-Cav (200-400 micrograms) and MIF-sLeu (200 micrograms) exerted antinociceptive effects in both tests. The coadministration of Kyo + L-NAME enhanced the nociceptive effect compared with L NAME (PP) or Kyo alone (PP, TF). The combination of Tyr-Cav + L-NAME enhanced the antinociceptive effect compared with L-NAME (PP) or Tyr-Cav alone (TF, PP). MIF-l (200 micrograms) had a weak antinociceptive effect in both tests. The coadministration of MIF-Cav + L-NAME enhanced the nociceptive effect compared with L-NAME (TF) or MIF-Cav alone (TF). The combination of MIF-sLeu + L-NAME enhanced the antinociceptive effect compared with L-NAME (TF) or MIF-sLeu alone (TF, PP). The results suggest that nitric oxide (NO) is involved in the antinociceptive effects of neuropeptides in the rat spinal cord. PMID- 12731455 TI - Selective mitochondrial KATP channel activation by nicorandil and 3-pyridyl pinacidil results in antiarrhythmic effect in an anesthetized rabbit model of myocardial ischemia/reperfusion. AB - The roles of cardiomyocyte sarcolemmal ATP-sensitive K+ (KATP) and mitochondrial KATP channels in cardioprotection and antiarrhythmic activity induced by KATP channel openers remain obscure. However, it has been suggested that the mitochondrial KATP channels are involved as a subcellular mediator in cardioprotection afforded by ischemic preconditioning. In the present study, we investigated the effects of the administration of non-hypotensive doses of ATP sensitive K+ channel (KATP) openers (nicorandil and 3-pyridyl pinacidil), a specific mitochondrial KATP channel blocker (5-hydroxydecanoate) and a specific sarcolemmal KATP channel blocker (HMR 1883; 1-[5-[2-(5-chloro-o-anisamido)ethyl] 2-methoxyphenyl]sulfonyl-3- methylthiourea) prior to and during coronary occlusion, as well as prior to and during post-ischemic reperfusion, on survival rate, ischemia-induced and reperfusion-induced arrhythmias and myocardial infarct size in anesthetized albino rabbits. The thorax was opened in the left 4th intercostal space and after pericardiotomy the heart was exposed. In Group I (n = 80), occlusion of the left main coronary artery and hence, myocardial ischemia induced arrhythmias were achieved by tightening a previously placed loose silk ligature for 30 min. In Group II (n = 186), arrhythmias were induced by reperfusion following a 20 min ligation of the left main coronary artery. In both Group I and Group II, early intravenous infusion of nicorandil (100 micrograms/kg bolus + 10 micrograms/kg/min), 3-pyridyl pinacidil (3.0 micrograms/kg bolus + 1.0 microgram/kg/min), HMR 1883 (3 mg/kg)/nicorandil and HMR 1883 (3 mg/kg)/3-pyridyl pinacidil, just prior to and during ischemia, increased survival rate (75%, 67%, 86% and 75% vs. 60% in the control subgroup in Group I; 67%, 75%, 75% and 67% vs. 43% in the control subgroup in Group II), significantly decreased the incidence and severity of life-threatening arrhythmias and significantly decreased myocardial infarct size. However, late intravenous administration of nicorandil or 3-pyridyl pinacidil at the onset of and during reperfusion did not increase survival rate nor confer any antiarrhythmic or cardioprotective effects. The antiarrhythmic and cardioprotective effects of both nicorandil and 3-pyridyl pinacidil were abolished by pretreating the rabbits with 5-hydroxydecanoate (5 mg/kg, i.v. bolus), a selective mitochondrial KATP channel blocker, but not by pretreatment with HMR 1883 (3 mg/kg). In the present study, higher levels of malondialdehyde (MDA) and lower levels of reduced glutathione (GSH) and superoxide dismutase (SOD) in the necrotic zone of myocardium in all sixteen subgroups in Group II suggest little anti-free radical property of nicorandil and 3-pyridyl pinacidil. Therefore, we may conclude that intervention by intravenous administration of nicorandil and 3-pyridyl pinacidil (through the selective activation of mitochondrial KATP channels), increases survival rate and exhibits antiarrhythmic and cardioprotective effects during coronary occlusion and reperfusion in anesthetized rabbits, when administered prior to and during coronary occlusion. The mitochondrial KATP channel may be considered to be a potentially important site of cardioprotection and antiarrhythmic activity. PMID- 12731456 TI - Efficacy of ebastine in the control of nasal congestion associated with allergic rhinitis. AB - Three similarly designed, multicenter, double-blind, randomized, placebo controlled, parallel-group comparative studies were carried out in the United States in a total of 1,881 patients to evaluate the efficacy of ebastine 20 mg (E20), ebastine 10 mg (E10), loratadine 10 mg (L10), and placebo (P), all given once daily, in controlling the symptoms of ragweed-induced rhinitis over a 4-week treatment period. Efficacy was assessed, among other means, by nasal congestion symptom scores entered by patients on diary cards in the morning and before bedtime over the previous 12-h period (reflective score, R) and at the time of recording (snapshot score, SS). Mean value of both morning and evening score changes from baseline were analyzed in each study and for each treatment. E20 was more effective than placebo in all studies, in both R and SS symptom scores (6 of 6 scores), while E10 was effective in 4 of 6 scores (2 R and 2 SS). In contrast, L10 was effective in only 1 of 6 scores (1 R). In conclusion, the comparative analysis of the results from these three trials shows that ebastine is efficacious in the reduction of nasal congestion associated with seasonal allergic rhinitis. This symptomatic effect of ebastine may be accounted for by its ability to reduce inflammatory markers, as shown in preclinical studies, in addition to its primary effect of antagonizing histamine H1 receptors. PMID- 12731457 TI - The effects of anticancer chemotherapeutic drugs on cognitive function and other neuropsychiatric dimensions in breast cancer patients. AB - This preliminary study aimed to apply a novel computerized measure derived from the content analysis of 5-min speech samples from patients with breast cancer to measure cognitive impairment and other neuropsychiatric dimensions during the course of anticancer chemotherapeutic treatment. Since such patients are often administered other pharmacological agents to alleviate their symptoms in addition to anticancer chemotherapeutic agents, another aim was to try to distinguish the mental effects of the anticancer drugs from the effects of any other drugs administered. Before and during the course of their anticancer chemotherapy, 12 breast cancer patients gave 5-min verbal samples, elicited by purposely ambiguous instructions, to talk about any personal life experiences. The recorded verbal samples were scored by a computer program (PCAD 2000) to measure the magnitude of cognitive impairment and other relevant neuropsychiatric dimensions. All of the pharmacological agents administered to the patients were recorded. The computer program automatically compared the scores derived from each verbal sample to already established norms to determine whether each score was within normal limits or one to three standard deviations from the norms. Significantly elevated Cognitive Impairment Scale scores were found in the verbal samples of 9 of the 12 patients. All patients had instances of elevated Health/Sickness Content Analysis Scale scores as well as frequent significantly elevated scores in shame anxiety and in death anxiety. In the Quality of Life Content Scale, the scores were uniformly low, ranging from +1.64 to -9.11. Further studies are being carried out to determine which patients are especially susceptible to cognitive impairment under these treatment conditions. PMID- 12731458 TI - The pharmacokinetic characteristics of levetiracetam. AB - Levetiracetam is the latest in a series of nine new antiepileptic drugs (AEDs) to be licensed for clinical use. Its present license is for use as adjunctive therapy for the treatment of patients with partial seizures with or without secondary generalization that are refractory to other established first line AEDs. Pharmacokinetic studies of levetiracetam have been conducted in healthy volunteers, in patients of all ages with epilepsy, and in certain special populations. Results of these studies indicate that levetiracetam has a very favorable pharmacokinetic profile, characterized by excellent oral absorption and bioavailability (> 95%) and a mean elimination half-life in adults, children and the elderly of 7, 6 and 10.5 h, respectively. Levetiracetam is not bound to plasma proteins and is not metabolized in the liver, so it is not expected to be associated with significant pharmacokinetic interactions. Indeed, to the best of the author's knowledge, no clinically relevant pharmacokinetic interactions with levetiracetam have yet been identified. However, pharmacodynamic interactions with carbamazepine and topiramate have been highlighted. As levetiracetam is primarily excreted unchanged in urine, dosage adjustments are necessary for patients with moderate-to-severe renal impairment. Overall, the pharmacokinetic characteristics of levetiracetam can be considered highly desirable. PMID- 12731459 TI - Therapeutic impact of CYP2C19 pharmacogenetics on proton pump inhibitor-based eradication therapy for Helicobacter pylori. AB - Current regimens for the eradication of Helicobacter pylori consist of a proton pump inhibitor (PPI) plus one or two antibacterial agents, such as amoxicillin (AMPC), clarithromycin (CAM) or metronidazole (MNZ). PPIs are mainly metabolized by S-mephenytoin 4'-hydroxylase (CYP2C19) in the liver. The polymorphism of CYP2C19 is associated with the pharmacokinetics and pharmacodynamics of PPIs. Eradication rates by PPI-based therapies are also affected by this genotype, as well as bacterial resistance to antibiotics. An individualized treatment strategy based on CYP2C19-related pharmacogenetics or pharmacogenomics and bacterial resistance is expected to increase the cure rate of the initial treatment. It is also necessary to recognize that there is a possible drug-drug interaction between some of the drugs used in this treatment regimen. PMID- 12731460 TI - Gateways to clinical trials. March 2003. AB - Gateways to clinical Trials is a guide to the most recent clinical trials in current literature and congresses. The data in the following tables has been retrieved from the Clinical Studies knowledge area of Prous Science Integrity, the drug discovery and devlopment protal, http://integrity.prous.com. This issue focuses on the following selection of drugs: AAV-CF, adalimumab, ademetionine, afeletecan hydrochloride, agomelatine, alemtuzumab, almotriptan, amdoxovir, aplidine, aranose, arsenic sulfide, atazanavir, atlizumab; Bimatoprost, BMS 181176, BMS-188667, bortezomib, bryostatin 1; Combretastatin A-4 phosphate; Darbepoetin alfa, darusentan, deferasirox, desloratadine, DTaP-HBV-IPV/Hib vaccine, DTI-0009; Eculizumab, edodekin alfa, emtricitabine, enfuvirtide, epoetin, esomeprazole magnesium etoricoxib; Fampridine, fenretinide, FR-146687; Galiximab, gamma-Hydroxybutyrate sodium, ganirelix acetate, gefitinib, Gemtuzumab ozogamicin, gimatecan; HEA125xOKT3, hIL-13-PE38QQR, HSV-2 theracine, Hu14.18-IL 2, human gammaglobulin; Idraparinux sodium, imatinib mesylate, IMiD3, insulin detemir, interleukin-4, irofulven, ISAtx-247; JT-1001; Levetiracetam, levosimendan, liposomal doxorubicin, liposomal vincristine sulfate, lixivaptan, lopinavir, lumiracoxib; Maxacalcitol, melatonin, midostaurin, MLN-518; Neridronic acid, nesiritide, nitronaproxen; Oblimersen sodium, oregovomab; PEG-filgrastim polyglutamate paclitaxel, prasterone, pregabalin; Rosuvastatin calcium, rotigotine hydrochloride; SGN-30; T-1249, tenofovir disoproxil fumarate, teriparatide, tiotropium bromide, tipranavir, TMC-114, trabectedin, transdermal selegiline; UK-427857; Valdecoxib, valganciclovir hydrochloride, vardenafil, vatalanib succinate, vincristine sulfate TCS; Zofenopril calcium. PMID- 12731461 TI - [Vitamin K deficiency bleeding in an infant despite adequate prophylaxis]. AB - Vitamin K deficiency in infants can cause life-threatening haemorrhages. To prevent this, neonates in the Netherlands receive an oral dose of 1 mg vitamin K directly after birth. In addition, because breast milk contains little vitamin K, breast-fed infants receive a daily dose of 25 micrograms the first three months. Of three female infants aged 4 weeks, 5 months and 3 months, respectively, two developed an intracranial haemorrhage, which caused death in one. In two cases there were signs of a bleeding tendency, but no tests were done because the patients appeared healthy otherwise. The underlying resorptive disorders, cholestasis and fat malabsorption, caused few symptoms and were discovered only after a vitamin K deficiency bleeding had occurred. In an infant with a bleeding tendency, one should consider the possibility of vitamin K deficiency, even if adequate prophylaxis has been given. PMID- 12731462 TI - ['No cure, no pay' in Dutch health care: fear of a culture of unwarranted claims]. AB - The Dutch Bar Association is considering the introduction of the contingency fee system ('no cure no pay'). Opponents believe that such a system will lead to an increase of legal procedures in health care as well as in other areas. However, experiences from the United States point in another direction. It appears that other elements of the Unites States tort system have a far greater risk of causing legalization than the contingency fee, notably the litigious society structure, the large compensations and the system of punitive damage, the jury system, and the widespread lack of social security. Nevertheless, a no fault health insurance system is preferable to the contingency fee system. PMID- 12731463 TI - [Atrial and B-type natriuretic peptides: from the research lab to clinical practice]. AB - The heart produces two related hormones, atrial (ANP) and B-type natriuretic peptides (BNP). Both are synthesized in the atria and ventricles as polypeptides, which upon release are split into ANP and BNP and the N-terminal fragments N-ANP and N-BNP (together named natriuretic peptides, NPs). The most important function of ANP and BNP is protection against volume-overload, by increasing natriuresis and diuresis amongst other things. Both peptides can be considered the natural antagonist of the renin-angiotensin system. Clearance occurs through a specific receptor and through enzymatic break-down by neutral endopeptidase (NEP). All 4 NPs circulate in plasma. Elevated concentrations of NPs are found when the filling pressures of the heart are elevated, as in acute coronary syndromes and congestive heart failure. Measurement of NPs is a useful tool in the differential diagnosis of cardiac versus non-cardiac dyspnoe (high negative predictive value), in the identification of heart failure patients most at risk and in optimising therapy in heart failure. In right ventricular overload caused by (corrected) congenital heart diseases and acute lung embolism, NP concentrations are also elevated, as they are in renal failure and in hypertension associated with left ventricular hypertrophy. Infusions of ANP and BNP lead to increased natriuresis and diuresis. Pharmacologically, increases in ANP and BNP can be accomplished with NEP-inhibitors or beta-blockers. Measurement of NP(s) will become as important for estimation of heart function as creatinine is for estimation of renal function. PMID- 12731464 TI - [Nutrition and health--vitamin supplementation and the vitality of the elderly]. AB - The increasing number of elderly people above 65 years of age is extremely heterogeneous with regard to sickness and health. As a result, the nutritional needs and food consumption also vary. 'Anorexia of aging', leading to nutritional deficiencies, is a common syndrome in the elderly people and may be either caused by or result in loss of functions and accelerated aging. Early detection of malnutrition and intervention with supplements or an adequate diet should stop the negative health spiral. A limited number of intervention trials, including Dutch ones, have demonstrated a positive effect on the nutritional and immune status. However, the improvement in physical and mental functions as a result of supplementation is less clear. The results of larger current trials on the effect of supplementation with, for example, vitamin D, folic acid, vitamin B12 and complete enriched products will provide more clarity in the future on the necessity of supplementation for elderly people. PMID- 12731465 TI - [Nutrition and health--vitamins and vitamin supplements]. AB - A balanced diet based on the Guidelines of the Netherlands Nutrition Centre provides a suitable basis for the maintenance of good health. However, there are a number of situations where supplementation with vitamins is clearly indicated. These include infants (vitamin A, D and K), young children, and pregnant and lactating women (vitamin D), future expectant mothers (folic acid) and the elderly (vitamin D). If doubts exist about a sufficient vitamin intake via the regular diet, a daily supplement supplying all vitamins at the level of the recommended daily allowance (RDA) is considered to be a responsible and safe choice. Epidemiological research indicates that the incidence of certain diseases is lower if the intake of vitamins is significantly higher than the RDA. However to date, targeted intervention studies have provided little unequivocal evidence to support this argument. For certain vitamins (A, D, folic acid, B6, nicotinic acid and beta-carotene) excessive intakes are associated with a health risk or clear toxicity. In the case of vitamin B6, nicotinic acid, folic acid and beta carotene this risk is mainly limited to the use of high-dose supplements. PMID- 12731466 TI - [Diagnostic image (135). A woman with acute abdomen. Volvulus of the small bowel due to strangulation]. AB - In a 79-year-old woman with acute abdomen CT revealed a 'whirl sign' configuration, pathognomonic for volvulus of the small bowel. PMID- 12731467 TI - [From gene disease; alpha1-antitrypsine deficiency]. AB - Alpha-1-antitrypsin (alpha 1-AT) deficiency is an autosomal, recessive hereditary disorder that is associated with the development of pulmonary emphysema and liver disease. The most common mutant leading to the deficiency is the Z mutant. This results in alpha 1-AT that forms polymers which lead to accumulation in the liver and decreased secretion. Emphysema is thought to occur due to insufficient protection of the lung tissue from the proteolytic activity of neutrophil-derived elastase, as a result of insufficient local alpha 1-AT levels. PMID- 12731468 TI - [Laparoscopic cholecystectomy in day care; implementation of a guideline for clinical practice]. AB - OBJECTIVE: To evaluate the extent to which the guideline to perform laparoscopic cholecystectomy in day care was followed in the institute where the guideline was developed and in the other hospitals in the Netherlands. DESIGN: Retrospective, descriptive. METHOD: For the patients who underwent a laparoscopic cholecystectomy in the Amsterdam University Medical Centre (AMC) during the period 1 January 1998-31 December 2000, data were collected from medical records on how the intervention was performed: in day care or during a hospital admission. The national figures were obtained from SIG Zorginformatie in Utrecht. RESULTS: A total of 262 patients underwent a laparoscopic cholecystectomy at the AMC: 60 men and 202 women, with an average age of 43 years (range: 2-86). A total of 163 (62%) patients were eligible for treatment in day care. Laparoscopic cholecystectomy was performed in day care in the years 1998, 1999 and 2000 in 80%, 92% and 93% of patients respectively (n = 144). Of these 144 patients, 129 (90%) were discharged to home the same day. For the same years, these figures for the whole of the Netherlands were 0.8%, 1.3% and 1.7% respectively. CONCLUSION: The clinical guideline for performing laparoscopic cholecystectomy in day care has been implemented to a large extent in the AMC but has scarcely been implemented at the national level. At the AMC, day care treatment could be carried out for 90% of the patients for whom this had been planned. PMID- 12731469 TI - [Lacrimal duct probing in young children with a congenital lacrimal duct obstruction at the Utrecht University Medical Center: generally an effective treatment]. AB - OBJECTIVE: To assess the results of lacrimal duct probing in young children with epiphora, due to a congenital nasolacrimal duct obstruction. DESIGN: Retrospective, descriptive. METHOD: Data were collected from medical dossiers on the results of lacrimal duct probing in children (0-48 months) with epiphora that was done in the period from January 1, 1997 to December 31, 2001 at the University Medical Centre in Utrecht, the Netherlands. The percentage of eyes that showed complete resolution of symptoms three months after the final probing was calculated. RESULTS: Of the 89 children who had undergone lacrimal duct probing, seven were excluded and in three children (six eyes) the data could not be retrieved. In 96 of the remaining 116 eyes (83%), the symptoms disappeared: this included 96% of the age group 0-12 months (n = 25), 85% of the 13-24 months olds (n = 55), 77% of the 25-36 months-olds (n = 22) and 57% of the 37-48 months olds (n = 14). No complications of probing were seen. CONCLUSION: In most children in the various age groups, epiphora disappeared. Thus, children do not need to be probed in the first year of life out of fear of failure at an older age. Whether or not probing shortens the duration of epiphora cannot be determined on the basis of either this study or the literature. PMID- 12731470 TI - [Non-traumatic myositis ossificans circumscripta in the teres major muscle]. AB - A 49-year-old male presented with a painful progressive swelling in his right axillar region, without further complaints, which had been present for 2 weeks. On radiological examination a peripheral circumferential zone of mineralisation was seen in the right teres major muscle. An incision biopsy specimen showed a lesion of fibroblastic tissue in which areas of osteoid and fragmented lamellar bone tissue, without signs of malignancy. The diagnosis was myositis ossificans circumscripta. This is a rare benign ossifying lesion in skeletal muscles, mostly caused by a trauma and with an average age of occurrence between 20 and 30 years old. It must be differentiated from extra-skeletal osteosarcoma. The pathogenesis is unknown. Because it is a benign and self-limiting disorder, surgical excision is only necessary in case of mechanical hindrance. The patient's swelling partially regressed and he had no further complaints. PMID- 12731471 TI - [Penis friction edema: not a venereal disease]. AB - A 35-year-old man presented with a local swelling of the penis, which increased until the entire penis was thick and swollen. After infectious and obstructive causes had been eliminated, a diagnosis of 'penis friction oedema' was made. The swelling disappeared during several weeks of abstinence from sexual intercourse. Penile abnormalities can be divided into venereal diseases (STDs), incidental affections of the skin or mucous membranes in that location, and disorders of vascularisation and lymph drainage. A traumatic disorder of lymph drainage is sometimes referred to, unjustifiably, as 'penile venereal oedema'; it is a result of friction and manifests itself as local or total penile oedema or as a cordlike congestion of the lymphatic vessels. The diagnosis is by exclusion and the treatment is temporary abstinence from sexual intercourse. PMID- 12731472 TI - [Asylum seekers in the Pacific Ocean: high prevalence of psychiatric disorders due to camp conditions]. AB - Australia deports its asylum seekers to poor neighbouring islands like Nauru in the Pacific. The asylum seekers are kept in detention on these islands, with serious consequences for their mental health and a high incidence of psychiatric disorders. Proper mental health care is virtually impossible under these conditions. Detention is unnecessary and inhumane. The International Organization for Migration, responsible for running these camps, should reconsider its mission as the world's leading organisation for migration issues. PMID- 12731474 TI - [The meaning of the sixth edition of "Organon der Heilkunst"]. AB - Like any artist, the paractitioner of the healing art should exactly know his tools, i.e. his medicines and the directions for their application. In addition, he should be able to rely on their genuineness. Regarding the tool "Organon", considerable uncertainty has been expressed by some physicians about the authenticity of Hahnemann's instructions for preparing and applying Q-potencies, as described in the sixth edition of the Organon of Medicine published by Richard Haehl in 1921. Since 1992, however, the first text-critical edition of the sixth edition of the Organonis available. Unlike Haehl's edition this one is exclusively based on Hahnemann's original manuscript and precisely transcribing all its handwriting, vouches for its authenticity. Hence there is no reason left to ignore Hahnemanns instructions concerning Q-potencies. This paper presents the history of the reception of Q-potencies as well as their prerequisites and evolution up to Hahnemann's final modifications of his earlier directions. As it turns out, these late instructions of Hahnemann do not mean complete change of all his previous opinions. Rather they are the logical completion of a course followed by him for ten years already. Q-potencies were Hahnemann's solution of the following therapeutic dilemma: on the one side physicians are inclined to repeat the dose of a high potency as often as possible in order to accelerate the process of healing; on the other side they should refrain from repeating the dose to avoid violent aggravations of the state of the patient. PMID- 12731473 TI - [Tuberculosis spondylodiscitis in three older patients]. PMID- 12731475 TI - Endocrine-related resources from the National Institutes of Health. PMID- 12731476 TI - [The golden calf and bronze snake. Concerning the purity of medical education]. PMID- 12731477 TI - Suggestions made for reducing cardiac drug errors. PMID- 12731478 TI - Syndromic diagnosis of reproductive tract infections leads to substantial unnecessary treatment in Vietnam. PMID- 12731480 TI - Although abortion is highly restricted in Cameroon, it is not uncommon among young urban women. PMID- 12731479 TI - The youngest children are too frequently subject to drug errors, study says. PMID- 12731481 TI - Focused interventions to help PCPs improve diabetic care don't produce lasting improvements. PMID- 12731482 TI - Bangladeshi women weigh a variety of factors when choosing a contraceptive. PMID- 12731483 TI - Genital cutting may alter, rather than eliminate, women's sexual sensations. PMID- 12731484 TI - New treatment option emerges for newly diagnosed multiple myeloma. PMID- 12731486 TI - Half of all adults with HIV are women. PMID- 12731485 TI - Offering a woman sterilization during an emergency cesarean section may sometimes be appropriate. PMID- 12731487 TI - HIV prevalence in South Africa. PMID- 12731488 TI - [Archeology of the radio pharmaceutical advertisement]. AB - After Second World War, a debate sets in France the partisans and the detractors of the radio advertisement, in particular pharmaceutical advertisement. In this article, the author revises campaigns led, during the thirties, by Robert Desnos for Armand Salacrou. PMID- 12731489 TI - Institutional review boards and approval of new products without human subjects research. PMID- 12731490 TI - Informed consent and living human cells. PMID- 12731491 TI - Hospital defendant says there was no basis for legal complaint. PMID- 12731492 TI - Institutional review boards and the privacy of human research subjects. PMID- 12731493 TI - Recruiting human subjects without IRB review/approval. PMID- 12731494 TI - IRBs and public use data files. PMID- 12731495 TI - Reporting problems in human research and resultant investigations by IRBs. PMID- 12731497 TI - Potential research subject complains about the violation of her privacy. PMID- 12731496 TI - Researcher accused of not reporting adverse events: U.S. v. John S. Najarian (Part 1). PMID- 12731498 TI - Institutional review boards (IRBs) would have an increased role in international research. PMID- 12731499 TI - Expansion of protections to all human research participants. PMID- 12731500 TI - Court is asked to invalidate the Food and Drug Administration's (FDA's) "pediatric rule". PMID- 12731501 TI - Financial conflicts of interest and human subjects research. PMID- 12731502 TI - Controlling desires: sexual orientation conversion and the limits of knowledge and law. PMID- 12731503 TI - Regulating managed care: what's wrong with a patient bill of rights. PMID- 12731504 TI - Translation of highly promising basic science research into clinical applications. AB - PURPOSE: To evaluate the predictors of and time taken for the translation of highly promising basic research into clinical experimentation and use. METHODS: We identified 101 articles, published between 1979 and 1983 in six major basic science journals, which clearly stated that the technology studied had novel therapeutic or preventive promises. Each case was evaluated for whether the promising finding resulted in relevant randomized controlled trials and clinical use. Main outcomes included the time to published trials, time to published trials with favorable results ("positive" trials), and licensed clinical use. RESULTS: By October 2002, 27 of the promising technologies had resulted in at least one published randomized trial, 19 of which had led to the publication of at least one positive randomized trial. Five basic science findings are currently licensed for clinical use, but only has been used extensively for the licensed indications. Promising technologies that did not lead to a published human study within 10 to 12 years were unlikely to be tested in humans subsequently. Some form of industry involvement in the basic science publication was the strongest predictor of clinical experimentation, accelerating the process by about eightfold (95% confidence interval: 3 to 19) when an author had industry affiliations. CONCLUSION: Even the most promising findings of basic research take a long time to translate into clinical experimentation, and adoption in clinical practice is rare. PMID- 12731505 TI - Incompetent organ donors. PMID- 12731507 TI - Little People of America: position statement on genetic discoveries in dwarfism (1996). PMID- 12731508 TI - Stem cells, embryos, and casualties of war. PMID- 12731511 TI - Health care rights: distinct claims, distinct justifications. PMID- 12731514 TI - Gas chromatography problem solving and troubleshooting. PMID- 12731512 TI - Immunosuppression, immunoisolation and cell therapy. PMID- 12731515 TI - Liquid chromatography problem solving and troubleshooting. PMID- 12731516 TI - Dark side of the genome: genetic blueprints can hinder as well as speed up medical research. PMID- 12731517 TI - Cancer scare hits gene cures: a second major setback for medicine's most pioneering treatment has split the scientific community. Could a moratorium do more harm than good? PMID- 12731518 TI - Your very own sequence. Last month, entrepreneur Craig Venter announced a bold new target: for anyone to be able to get their genome sequenced for under $1000. So can it be done? And what use would it be knowing your genome sequence anyway? PMID- 12731519 TI - Birds, bees and laser beams: the idea of parents choosing the sex of their children is no longer science fiction. PMID- 12731520 TI - Genes can come true. PMID- 12731521 TI - Organ transplantation: the ethics of consequences. PMID- 12731522 TI - The new reproductive technologies: an overview and theological assessment. PMID- 12731524 TI - Being a Catholic physician at the beginning of the third millennium. PMID- 12731523 TI - Pragmatism and conscience: a religious perspective on competing values in embryo experimentation. PMID- 12731525 TI - Physician-assisted suicide: the ultimate Freudian legacy. PMID- 12731527 TI - Human life as a foundation for ethical health care decisions: a synthesis of the work of E.D. Pellegrino and W.A. Wallace. PMID- 12731526 TI - On rescuing frozen embryos. PMID- 12731528 TI - The morality of human embryonic stem cell research and President Bush's decision: how should Catholics think about such things? PMID- 12731530 TI - Cloning, stem cell research and some historic parallels. PMID- 12731529 TI - Why respect for the human embryo? PMID- 12731531 TI - How many lawyers does it take...greed and legal wrangling could stifle the medical revolution. PMID- 12731532 TI - Footing the bill: should we all have to pay for one company's bright idea? PMID- 12731533 TI - Say no to stem cell patents: Europe advised to take a different stance to the US. PMID- 12731534 TI - A tale of two youngsters: two boys, two diseases. One will be treated, one won't. Why? PMID- 12731535 TI - There's no pill for curing a hellish upbringing. PMID- 12731536 TI - Childhood is not what it used to be: the rise of Ritalin has more to do with shifting social values than advances in neuropharmacology. PMID- 12731537 TI - Australia agonises over stem cells. PMID- 12731538 TI - Who's to say that the risk is worth taking? Faced with the fear that their science might be misused, what should a researcher do? PMID- 12731539 TI - Brain teaser: changing people's behaviour by implanting electrodes in their brains raises a host of troubling questions. PMID- 12731540 TI - Only as a last resort: primate experiments are justified--if scientists can tell us what they're for. PMID- 12731541 TI - The DNA helix at 50. PMID- 12731542 TI - Animal experiments on trial. PMID- 12731543 TI - Corixa: integrating immunotherapeutics. PMID- 12731544 TI - I see a long life and a healthy one... PMID- 12731545 TI - Genetic testing for cystic fibrosis: a personal perspective. PMID- 12731547 TI - Bibliography. Current world literature. Neuroscience. PMID- 12731546 TI - Foreward: genetic exceptionalism. PMID- 12731548 TI - Bibliography. Current world literature. Gastrointestinal system. PMID- 12731549 TI - DNA: five distinguishing features for policy analysis. PMID- 12731551 TI - Does the FDA have authority to regulate human cloning? PMID- 12731550 TI - Confused heritage and the absurdity of genetic ownership. PMID- 12731552 TI - Is there a right to clone? Constitutional challenges to bans on human cloning. PMID- 12731553 TI - Parpalaix v. CECOS: Protecting Intent in Reproductive Technology. PMID- 12731554 TI - "...And this little pig saved lives!". AB - The harvesting and transplantation of organs from genetically altered pigs has sparked an escalating debate within the transplant community worldwide. The ethical and moral arguments against using another life form to save humans--as well as using humans for experimentation--run side by side with the potential medical considerations of inadvertently unleashing heretofore unknown virulent agents on the public. Nonetheless, the ethical case against xenografting, while compelling and worthy of more public and expert debate and discussion, is not persuasive. Genetically altered pigs might not be able to save human lives at present, but the time has come to take a tentative step to see whether they can in the future. PMID- 12731555 TI - Viewing health care as a common good: looking beyond political liberalism. PMID- 12731556 TI - The legitimization of fetal tissue transplantation research under Roe v. Wade. PMID- 12731557 TI - Genetic privacy: new intrusion a new tort? PMID- 12731559 TI - Constitutional persons: an exchange on abortion. PMID- 12731558 TI - Position of the American Dietetic Association, Society for Nutrition Education, and American School Food Service Association: Nutrition services: an essential component of comprehensive school health programs. PMID- 12731560 TI - Cloning human embryos for spare tissue: an ethical dilemma. PMID- 12731562 TI - Bio-policy and the place of institutionalised ethics in political decision making. AB - Questions concerning moral problems caused by the life-sciences and concerning the adequate methods and instruments to solve these are timely and urgent; especially in the face of intense debates on the acceptability of research on human embryonic stem cells and preimplantation diagnostics, to name only two applications developed from research on the life-sciences. Unfortunately, the constant and accusing demand that life-scientists must behave morally does not give us a clue on how ethics may help in establishing guidelines for moral behaviour. In this heated situation the foundation of ethics-committees seems to be the motto of the day. But instead of functioning as necessary and fruitful scientific advisory boards, these committees run the risk of being misused as a fashionable (and soon forgotten) weapon in the battle for political opinion leadership. In the following article, the view is defended that ethics is a scientific enterprise and has an important role to play in political decision making on life-sciences issues. PMID- 12731561 TI - Legal responses to some of the new developments in reproductive technologies. Part 3: the future of reproductive technologies and the law. PMID- 12731563 TI - Bioethical dilemmas through patristic thought. PMID- 12731564 TI - Bioethics and human rights. PMID- 12731565 TI - Bibliography. Current world literature. Pediatrics. PMID- 12731566 TI - Bibliography. Current world literature. Valvular heart disease. PMID- 12731567 TI - Bibliography. Current world literature. Heart transplantation. PMID- 12731568 TI - Stimulation of lymphocyte proliferation by non-lymphoid porcine tissue cells. AB - Cultured porcine non-lymphoid cells, characterized by biochemical and morphological criteria, were derived from different tissues of individuals typed by serological and mixed lymphocyte culture methods for gene products of the major histocompatibility complex. These cultured cells have been used as stimulators in mixed lymphocyte-tissue cell cultures in order to investigate (1) the magnitude, kinetics and dose-dependence of lymphocyte transformation caused by tissue cells compared with that caused by lymphocytes as stimulators; (2) the relationship between the expression of serologically detected Ia-like antigens by tissue cells and their ability to cause lymphocyte transformation; (3) the genetic control of stimulation by tissue cells and by lymphocytes and (4) the expression and genetic control of lymphocyte stimulatory properties restricted to tissue cells and absent from lymphocytes. It has been shown that some but not all kinds of tissue cells can stimulate allogeneic lymphocytes strongly and that the characteristics of such stimulation are similar to those observed in mixed lymphocyte cultures. Strong stimulation by tissue cells does not always correlate with the expression of serologically detectable Ia-like antigens, but appears to be controlled by the major histocompatibility complex. There is evidence that certain tissue cells possess lymphocyte stimulatory properties not shared by lymphocytes. Preliminary data suggest that such tissue cell specific stimulation is not controlled by the major histocompatibility complex, though more detailed genetic analysis is required. PMID- 12731569 TI - Ankylosing spondylitis--the role of HLA-B27 homozygosity. PMID- 12731570 TI - Preparation of liposomes incorporating membrane components from human lymphoid cells. AB - A procedure for the preparation of liposomes incorporating membrane components of human lymphoid cells is described, whereby the detergent used to solubilize the plasma membrane is almost completely eliminated. MHC products HLA-A-B-C and DR molecules were shown to be incorporated into the liposomes and their antigenic determinants to be exposed on the outer surface. A protein-rich liposomal fraction could be separated from both protein-free liposomes and from aggregates. This preparation may be suitable for testing the functions of cell membrane components in biological assay systems. PMID- 12731571 TI - Measurement of cell mediated cytotoxicity by post-labeling surviving target cells. AB - The 51Cr release assay (CRA) is the commonly accepted technique for measurement of cell mediated cytotoxicity. This assay shows some disadvantages when mononucleated cells of human peripheral blood (MNC) are used as effector and target cells. The uptake of 51Cr by PHA stimulated lymphocytes is low compared to the spontaneous release. In an attempt to develop a cytotoxicity assay suitable for human lymphocytes we used 14C-TdR to label target cells surviving after contact with effector cells. Cytotoxic lymphocytes were generated by incubation of MNC with irradiated allogeneic MNC for 6 days. On day 6 the effector cells are irradiated and co-cultured with PHA stimulated target cells. Twenty-four hours later 14C-TdR is added. After an additional 24 h the cultures are harvested and 14C-TdR taken up by target cells is measured. It is shown that the effector cells are still cytotoxic after irradiation. These cells do not take up 14C-TdR. Cell free supernatants do not influence the uptake of 14C-TdR by target cells. The results obtained with this assay correlate very well with those obtained by the CRA, if the spontaneous release does not exceed 30%. PMID- 12731572 TI - The major histocompatibility complex (HLA) as a genetic marker in human craniofacial anomalies. AB - Study of the incidence and segregation of the serologically detectable A and B products of the HLA complex in 140 family units in which one or more offspring was afflicted with a developmental craniofacial anomaly has uncovered no evidence of an association between HLA-A or B antigens or haplotypes and the malformations under study. Further analysis of HLA-D products in the same family units by the mixed leukocyte culture (MLC) technique has, however, uncovered a relatively high incidence of non-reactivity between the cells of one (or both) parent(s) and cells of some offspring in 41 of the 140 families included in this study. The parent couples involved in this finding were unrelated and generally did not share any HLA-SD haplotypes. When this finding was studied further by Primed LD Typing techniques, the results in six families suggested that such MLC non reactivity is a consequence of the sharing of LD alleles by each pair of parents in these families. The known polymorphism of the HLA-D locus (or loci) and the low incidence of comparable findings in the normal population suggest that LD allele sharing in this particular population may be related to the selection of certain particular HLA-D products in families afflicted with developmental craniofacial anomalies. This result may be relevant to the possible existence in man of an analogue of the murine T/t complex which may occur in linkage with the HLA complex, in the same manner as the linkage disequilibrium which is been documented between the t complex and H-2 in chromosome 17 of the mouse. PMID- 12731573 TI - Esophageal cancer studies in the Caspian Littoral of Iran: introductive assessment of the HLA profile in patients and controls. AB - The possibility of an association between HLA profile and the probability of having esophageal cancer was explored among patients from different ethnic groups residing in the Caspian Littoral of Iran. A total of 151 esophageal cancer patients and 214 normal controls with ethnic affiliations similar to those of the patients were typed for HLA antigens. No statistically significant differences were found between patients and controls with regard to HLA antigens. PMID- 12731574 TI - HLA-Dw antigens in unrelated juvenile, insulin-dependent diabetics. AB - Forty-one unrelated juvenile, insulin-dependent diabetics have been HLA tissue typed for A, B and Dw anitgens and compared with a normal control population. We have found statistically significant increases in the frequencies of B8, B18, and Dw3, and significant decrements in the frequencies of B7, B12 and Dw2. The log linear modeling technique was used to study the association of JIDD with Dw3 and B8 antigens. We confirmed that the B8 excess seen in diabetics is secondary to the excess of Dw3. The decrements of B7, B12 and Dw2 could reflect an association of these antigens with a protective factor for the disease, or could be due to an artifact. The latter possibility was excluded for B7 and Dw2 by adjusting for the excess antigen frequencies. These findings suggest that the associations between the HLA and diabetes are compatible with the existence of genes which are concerned with the pathogenesis of the disease and are closely associated with the D locus of the major histocompatibility system. PMID- 12731575 TI - Alloantibodies that react with subsets of human T cells. AB - Using the two color fluorescence (TCF) method, alloantibodies against subsets of T cells could be detected in sera from pregnant women with strong HLA antibodies. To preclude interference of these HLA antibodies with the recognition of the T cell antibodies, serum donors were selected which were HLA-Al, -B8, -DRw3. Their sera were tested on a panel of individuals homozygous for HLA-Al, -B8, -DRw3. By enriching peripheral mononuclear blood lymphocytes for Tgamma cells it could be shown that some of the sera reacted mainly with Tgamma and others with Tmu lymphocytes, while some sera reacted with both. PMID- 12731576 TI - HLA-B5 and Behcet's disease. PMID- 12731577 TI - Primary biliary cirrhosis associated with HLA-DRw3. AB - Twenty-one unrelated Caucasian patients with Primary Biliary Cirrhosis were typed for HLA-A, B, C and HLA-DRw antigens. The antigens HLA-Al, HLA-B8 and HLA-DRw3 were found in increased frequency in relation to the control group. When the P values were corrected for the number of antigens tested only the increase of HLA DRw3 remained significantly different from the control group (P crr. < 0.004). The antigen HLA-DRw3 was carried by 12 out of 21 patients (57.1%) in comparison to 11 out of 74 (14.8%) normal unrelated Caucasian subjects. PMID- 12731578 TI - A monoclonal mouse anti-rat Ia antibody which cross reacts with a human HLA-DRw determinant. AB - A mouse monoclonal antibody, called MRC OX 3, which detects a polymorphic Ia determinant in the rat and cross reacts with an Ia determinant coded for by the I A subregion in the mouse, detects a polymorphic determinant on human B cell lines. MRC OX 3 antibody binds to human B cell lines which express HLA-DRw specificities DRw 1, DRw2 and DRw6 but does not bind to those cells which only express other HLA-DRw specificities. No significant binding of MRC OX 3 antibody to either normal or mitogen stimulated human peripheral blood leukocytes, some of which are typed as HLA-DRw1, DRw2 or DRw6, could be detected. The binding of MRC OX 3 antibody to a human B cell line could be completely inhibited by pre absorbing the antibody with purified rat Ia antigen. PMID- 12731580 TI - Evaluation of commercial antiglobulin sera over a two-year period. Part I. Anti beta 1A, anti-alpha 2D, and anti-beta 1E levels. AB - Antiglobulin sera from nine different manufacturers have been tested, over a two year period, for their ability to detect the complement components beta 1A, alpha 2D, and beta 1E. The results demontrate considerable variation in the abilities of sera from different manufacturers to detect these components and indicate that not all sera on the market are suitable reagents for diagnostic use and compatibility tests. The results also show that there is considerable variation between different lots of serum from some of the manufacturers. In general, the anti-complement levels of these reagents have increased during the two-year period of study but not all companies produce suitable reagents for routine use. PMID- 12731579 TI - No significant association between HLA and nickel contact sensitivity. PMID- 12731581 TI - Evaluation of commercial antiglobulin sera over a two-year period. Part II. Anti IgG and anti-IgM levels and undesirable contaminating antibodies. AB - Antiglobulin sera, from nine different manufacturers, have been tested over a two year period for their ability to detect different nonagglutinating, IgG and IgM blood group antibodies and for the presence of undesirable antibodies that cause the agglutination of nonglobulin coated red blood cells. There are not so many differences in anti-IgG levels among the sera as there are differences in anticomplement levels. Over the two-year period, there have been, in general, increases in the amounts of anti-IgM antibodies in the sera tested. Several of the sera, apparently prepared by dilution of raw rabbit serum and not by adsorption, contain antibodies that cause the agglutination of red blood cells not coated with IgG, IgM, or any of the components of complement. PMID- 12731582 TI - Coagulation studies after transfusion of hydroxyethyl starch protected frozen blood in primates. AB - This-study evaluates the effects of the transfusion of frozen-thawed blood protected by hydroxyethyl starch (HES) on shocked monkeys. Particular attention is directed toward the study of coagulation in view of previous reports of coagulopathies caused by hydroxyethyl starch. In vivo studies were done in 13 shocked monkeys that received either homologous or autologous blood which had been frozen and thawed with 14 per cent HES as the cryoprotective agent. Multiple coagulation studies were obtained on the monkeys, and histopathological evaluations were performed. There was no evidence of increased bleeding or bleeding tendencies in the monkeys transfused with HES preserved blood. Serum chemistry values were not significantly altered in animals that received blood preserved by HES nor in those that received large quantities of HES alone. Histopathological examination of biopsy and autopsy material demonstrated no changes which could be related to hydroxyethyl starch. PMID- 12731583 TI - Bacteriocidal properties of platelet concentrates. AB - Platelet concentrates were contaminated with predetermined numbers of bacteria isolated from patients. Staphylococcus epidermidis and Pseudomonas aeruginosa organisms were killed if the number introduced was less than 10(3). When 10(4) through 10(6) organisms were added, their growth was depressed for 24 hours and then slowly resumed. If more than 10(7) organisms were used, a slow and progressive growth occurred. After the concentrates were contaminated with Staphylocoecus aureus, slow progressive growth occurred regardless of the size of the inoculum. Cultures taken from sample phlebotomies performed on a few donors showed that the number and type of organisms released into the blood during the venepuncture are such that they could usually, but not always, be inactivated by the bacteriocidal property of the platelet packs. This bacteriocidal property will usually allow the platelet packs to be stored at room temperature for considerable amounts of time with relative safety. Bacterial growth will normally occur only when the number of organisms introduced during the venepuncture is quite large, and growth will usually occur during the first 24 hours. PMID- 12731584 TI - Automated blood typing of patients. AB - The 15-channel Technicon blood typing machine is shown to be equally useful for patient and donor blood typing. It provides increased accuracy as well as increased speed and economy. Test capabilities are broadened by permitting additional simultaneous determinations such as the automated reagin test for syphilis or tests for additional erythrocyte antigens. Automated antibody screening, performed on the blood typing machine in conjunction with patient typing, was found to be of limited use because it failed to detect as much as 60 per cent of some clinically significant antibodies. PMID- 12731585 TI - The platelet response to hypotonic shock. Its value as an indicator of platelet viability after storage. AB - The nature of the platelet response to osmotic shock and its relationship to platelet viability were studied. Light absorbancy changes of human platelet concentrates exposed to hypotonic shock were measured in a spectrophotometer: a sudden drop of light absorbancy was followed by a reversal of light absorbancy towards normal (reversal reaction). It was confirmed that the reversal reaction is a complex phenomenon dependent on the integrity of biochemical and enzymatic functions of the platelets. It was suppressed by glycolytic inhibitors and by SH blocking agents. Ouabain had no immediate effect, but with prolonged incubation it depressed the reaction. Suspension of the platelets in a protein-free medium caused a rapid loss of the reversal reaction. Disappearance of the marginal bundle of microtubules by exposure to colchicine did not change the reaction leading to the hypothesis that microfibrils rather than the microtubules may have been responsible for the reversal reaction. The conclusion was derived that the reversal reaction is due to cell volume contraction for which integrity of the platelet contractile protein and energy availability are essential. Platelet storage at 4 degrees C or at 22 degrees C caused a progressive depression of the reversal reaction which was more severe in platelets preserved at 4 C than in those preserved at 22 degrees C, and paralleled the loss of the platelet capacity to survive in vivo. Cryoprotective agents (DMSO, DMAC and glycerol) partially inhibited the reversal reaction. Freezing with these agents caused a more severe depression of the reaction. The least depression was observed with 5 per cent DMSO. The results demonstrated that the reversal reaction is a valid and accurate in vitro indicator of in vivo platelet viability when the results to be compared are limited to a single method of storage. Usefulness of the reversal reaction is reduced when results obtained with different methods of storage are compared. PMID- 12731586 TI - Platelet preservation by freezing. Use of dimethylsulfoxide as cryoprotective agent. AB - Variables important in the preservation of platelets by freezing with dimethylsulfoxide (DMSO) as cryoprotective agent were studied in normal volunteers and thrombocytopenic patients. Use of 5 per cent DMSO and a freezing rate of 1-3 degrees C/minute yielded optimal preservation of platelet viability. The addition of 5 per cent Dextrose did not improve results. In vivo yield using 5 per cent DMSO was superior to previous results in which glycerol was used as the cryoprotective agent. Viability after freezing was equivalent when platelets were frozen in small (10 ml) and large (60 ml) volumes of plasma. The larger volume had the advantage that a smaller percentage of the platelets was lost during transfer from one plastic container to another. PMID- 12731587 TI - Cooling mattress induced acute hemolytic anemia. AB - We have reported a patient with Mycoplasma pneumonia, cold agglutinins, and a hemolytic anemia which was probably secondary to vigorous treatment of hyperpyrexia by a cooling mattress. Physicians caring for patients with Mycoplasma pneumonia should be alerted to the potential untoward effect of a cooling mattress on an individual with a high titer of cold agglutinins. PMID- 12731588 TI - American National Red Cross experience with hepatitis-B antibody (HBAb or anti HBAg) testing. AB - Blood collected by 11 Red Cross Regional Blood Centers were screened by the method of agar gel diffusion (AGD) for the presence of hepatitis-B antibody. Of the 185,134 units tested, 114 were found to be positive for HBAb by the Regional Centers and were forwarded to National Special Projects Laboratory for confirmation. Only five out of 114 samples revealed lines of identity with a control anti-HBAg when reacted with a pool of plasma positive for hepatitis-B antigen that was different from that which was provided to the Centers. Apparently, the precipitation reactions observed by the Centers were largely due to the antithrombin antibodies in the donors' sera reacting with the residual thrombin used by the manufacturer to convert HBAg positive plasma to serum. We conclude that the incidence of hepatitis-B antibody as measured by agar gel diffusion in the Red Cross Blood donor population was extremely low. PMID- 12731589 TI - Microaggregates in frozen and saline washed red blood cells. AB - The microaggregates which accumulate in stored blood have been implicated in the development of posttraumatic pulmonary insufficiency. These particles are known to be composed of degenerated leukocytes and platelets. Because frozen and saline washed red blood cells contain small numbers of leukocytes and platelets, they were studied as a possible source of microaggregate-free red blood cells. Using a Model T Coulter Counter to quantitate all particles 13-80 microns in size, it was shown that freezing and deglycerolization, or simple saline washing (manual or automated), could reduce the number of microaggregates in stored blood by 80 to 90 per cent. These findings add to a growing list of potential advantages in the routine use of frozen red cells for patients requiring transfusion. PMID- 12731590 TI - Clotting factors in supernatant plasma following cryoprecipitation. AB - The supernatant plasma which remains after cryoprecipitation is a useful component for transfusion therapy. However, from prior studies it was unclear how well plasma clotting factors were preserved following cryoprecipitation. Coagulation factor assays were performed on fresh plasma, supematant plasma after removal of cryoprecipitate, and the same supernatant plasma following refreezing and thawing. Factor VIII and fibrinogen levels fell considerably after removal of cryoprecipitate, but there was also a significant decline in factor V. Refreezing and thawing the supernatant plasma had little effect on clotting factors. PMID- 12731591 TI - A study of weak subgroups of blood group A with an antiglobulin-latex test. AB - The distribution of the blood group A antigen on subgroups A1, A2, A3, Ax, and Ae1 was studied by means of a highly sensitive and specific antiglobulin-latex test. Marked variability in the cell to cell expression of A antigen was observed in A1 and A2 cells. The weaker subgroups (A3, Ax, and Ae1) were characterized by a minor population of cells containing A antigen and a major population with no detectable A sites. Comparison of the antiglobulin-latex test with the fluorescent antibody technique indicates that the former is approximately 10,000 times more sensitive in detecting cell bound antibody, and thereby membrane antigens. The results of this study demonstrate that within each of the A subgroups (with the possible exception of A1) there exist erythrocytes that apparently lack recognizable A antigen sites. The number of such cells is greatest in the weakest subgroups (Ax and Ae1). PMID- 12731592 TI - Procurement and identification of HL-A lymphocytotoxic antibodies in sera of nonpregnant, multiparous blood donors. AB - An efficient procedure for large-scale screening of nonpregnant multiparous blood donor sera is described. Yield of useful HL-A reagents in this population compares favorably to yields obtained from screening pregnant women near term. The advantages of screening nonpregnant donors include stable antibody titers, ease of procurement, and the willingness of voluntary blood donors to return for plasmapheresis. PMID- 12731593 TI - Platelet concentrates: sterility of 400 single units stored at room temperature. AB - Four hundred single platelet concentrates were prepared and stored at room temperature for 72 to 96 hours. Triple cultures of each concentrate at varying incubation temperatures revealed only four positive cultures. The Corynebacterium species and Staphylococcus species, coagulase negative, were the only organisms identified and were regarded as contaminants. No transfusion reactions of bacterial origin were observed in transfusing 10,024 other single platelet concentrates. Room temperature preparation and storage of single platelet concenrates is a safe and practical procedure. PMID- 12731595 TI - Telling the truth. PMID- 12731596 TI - Improved design of long-span resin-bonded fixed partial dentures: three case reports. AB - When single-tooth implants are not appropriate, the use of resin-bonded fixed partial dentures is a preferred treatment option when the abutments are relatively sound. However, the use of resin-bonded fixed partial dentures (RBFPDs) for replacing two or more missing teeth is considered to have a guarded prognosis, as long-span RBFPDs have been shown to be less successful than single pontic prostheses. The use of properly modified nonrigid connectors may well improve the success of long-span RBFPDs by reducing harmful interabutment stresses that appear to be responsible for retainer debonding in long-span RBFPDs. For such long-span prostheses, it is advised that the major retainer have wraparound on at least three surfaces of the abutment or have strategically placed opposing axial grooves or slots. It is considered essential that the connector allow interabutment movement in both the horizontal and vertical planes so that the retainer with the greater resistance and retention form does not stress and possibly debond the minor retainer. The upside-down positioning of the nonrigid connector and the matrix incorporated with the major retainer is considered important for successful maintenance, if a debond should occur due to greater loading on the major retainer, because it can be removed and recemented easily. Clinical cases are described that replace two or more missing teeth using fixed-movable RBFPDs with nonrigid connectors. PMID- 12731594 TI - A checklist of American transfusion books: 1909-1972. PMID- 12731598 TI - Impressions for complete dentures using new silicone impression materials. AB - This article describes a convenient technique for making impressions of complete dentures using two newly developed silicone materials. One of these materials, a heavy-bodied silicone material, is used for simultaneous molding of all borders. The material is available in automatic mixing systems, so it can be easily and evenly applied on the tray borders. The material is designed to have a low elasticity after setting so that excess material can be carved or deficient sites corrected with a small mix. The other newly developed material, a light-bodied silicone material, possesses better flow than the usual light-bodied silicones. In addition, since viscosity is controlled and adequate flow is maintained during seating in the mouth, mucosal detail is superior. PMID- 12731597 TI - Influence of silanization on early bond strength to sandblasted densely sintered alumina. AB - OBJECTIVE: The increased popularity of alumina-based restorations has resulted in an interest in proper adhesive techniques to assure a strong and predictable bond to these restorations. This study investigated the early bond strength of three different resin-cement systems to densely sintered alumina (aluminum-oxide ceramic) with and without the use of their corresponding silane coupling agent (silanization). METHOD AND MATERIALS: Ninety samples of densely sintered high purity aluminum-oxide ceramic were randomly divided into three groups. Composite cylinders were bonded to the ceramic samples with three resin-cement/bonding agent systems: Noribond DC (NOR), Panavia 21 EX (PAN), and Variolink II (VAR). Each resin-cement/bonding-agent system was used with and without their corresponding silane (SIL) coupling agent (n = 15). After fabrication, the specimens were stored in distilled water for 3 days at room temperature, and shear bond strength was tested. RESULTS: Application of the silane-coupling agent on sandblasted densely sintered alumina did not significantly influence bond strengths with PAN. Silanization significantly improved shear bond strengths with NOR and VAR. NOR-SIL and VAR-SIL revealed the statistically highest values, with NOR-SIL showing the highest mean bond strength of all groups. CONCLUSIONS: Silanization of sandblasted densely sintered alumina had mixed effects on the applied resin cements: It had no effect on the performance of the phosphate modified resin cement PAN, but significantly improved shear bond strength of the Bis-GMA composite resin cements VAR and NOR. NOR-SIL revealed the highest overall mean bond strength. PMID- 12731599 TI - Quantitative light-induced fluorescence (QLF)--a potential method for the dental practitioner. AB - The quantitative light-induced fluorescence (QLF) method is based on the natural fluorescence of teeth. Currently, the method is predominantly used in vitro and in clinical studies for early detection of carious lesions and for monitoring of de- and remineralization of white spots by quantifying the mineral loss and the size of smooth surface lesions. The objective of this paper was to demonstrate the potential of QLF for application in routine dental care. Experience with QLF was reported by selected cases, which enlightened the advantages of the method for caries detection and for evaluation of preventive interventions in caries prone patients in daily practice. PMID- 12731600 TI - Three-dimensional analysis of dual-arch impression trays. AB - OBJECTIVE: The objective of this investigation was to measure and compare the dimensions of casts made using four types of impression trays and two impression materials to the dimensions of the original master model. This study differed from previous studies by using three-dimensional measurement. METHOD AND MATERIALS: Vinyl polysiloxane and polyether impressions were made of two crown preparations of ivorine teeth cemented into an acrylic master model. Two grooves were placed into each of the preparations. In addition, six points were placed on each occlusal surface and six were placed on each finish line. Impressions were poured with a Type IV dental stone at 48 hours. A three-dimensional measuring system was used to determine coordinates of 32 points on the master model and resulting casts. Inter- and intratooth dimensions were calculated from the measured coordinates. RESULTS: The accuracy and precision of the impression techniques were assessed using analysis of variance (ANOVA) and Dunnett's t test. The ANOVA and Dunnett's t test indicated that only the custom tray did not detect any inaccuracies and was as reproducible as the master model. CONCLUSION: This study suggests that the custom tray technique is most accurate, even though all other techniques evaluated apparently produce clinically acceptable results. PMID- 12731601 TI - A comparison of the retention of tooth-colored posts. AB - OBJECTIVE: The aim of this in vitro study was to compare the retention of five different esthetic post systems of similar dimensions in extracted teeth using titanium posts as controls. METHOD AND MATERIALS: Sixty recently extracted single rooted, caries-free teeth were sectioned horizontally and mounted in acrylic resin. The samples were randomly allocated into six groups of 10 for post preparation. Post space preparation was carried out according to the individual manufacturer's instructions. All posts were bonded using Panavia F. A 4-mm hollow, metal sleeve was luted over the free end of each post prior to mounting in a universal testing machine, and the forces required to dislodge the posts using a cross-head speed of 5 mm/min were recorded. RESULTS: It was found that the parallel-sided Lightposts were significantly more retentive than all of the other posts. Parapost Fibrewhite posts were more retentive than tapered Lightposts and Snowposts. There was no significant difference between the retention of stainless steel Paraposts and any of the other groups. CONCLUSIONS: Serrated parallel-sided stainless steel posts were no more retentive than either parallel-sided or tapered tooth-colored posts in this study. When all groups were considered, post dimension appeared to influence retention, with parallel-sided posts being more retentive than tapered posts. PMID- 12731602 TI - A new rubber dam frame design--easier to use with a more secure fit. AB - The application of the rubber dam (dental dam) is indicated in endodontics and for restorative dental treatments involving the acid-etch technique. The frequency of the technique's use varies significantly according to individual circumstances, and is generally too low given its advantages. One possible reason for many dentists' reluctance to use it could be frustrating results in the past with technically difficult applications. Such conditions arise, for example, when the taut rubber dam sheet exerts too much pull on the rubber dam clamps, causing them (and the entire rubber dam) to come loose. Particularly susceptible here are clamps attached to molars. This undesired tension results from stretching the rubber dam material--a necessary step--for attaching the sheet to the traditional rubber dam frame. As an alternative, a new easy-to-use rubber dam frame (Safe-T Frame) has been developed that offers a secure fit without stretching the rubber dam sheet. Instead, its "snap-shut" design takes advantage of the clamping effect on the sheet caused when its two mated frame members are firmly pressed together. In this way the sheet is securely attached, but without being stretched. Held in this manner, the dam sheet is under less tension, and hence, exerts less tugging on clamps--especially on those attached to molars. Even in cases where there are no distinct anatomic undercuts, this lack of tension in the sheet eases isolation procedures and permits the use of standard rubber dam clamps. As a further benefit, the frame's raised edging provides a barrier around the sheet, which prevents small amounts of fluids from escaping. This contributes to greater patient comfort. PMID- 12731603 TI - Extensive pneumatization of the temporal bone and articular eminence: an incidental finding in a patient with facial pain. Case report and review of literature. AB - A 53-year-old Caucasian female presented to the Oral Medicine Department at the hospital of the University of Pennsylvania for consultation regarding facial pain. A panoramic radiograph revealed multilocular radiolucencies in the right articular eminence. A CT scan was then performed, and the radiolucencies were determined to be pneumatization of the articular eminence. PMID- 12731604 TI - Adherence to periodontal maintenance in Tehran, Iran. A 7-year retrospective study. AB - Periodontal disease is chronic but not life threatening. Therefore, adherence to periodontal maintenance (PM) is often not ideal. However, this service is frequently needed for the remainder of one's life in order to prevent disease progression and recurrence. This study evaluates the degree of compliance with PM in a private periodontal practice in Tehran, Iran. Data were collected and analyzed for degree of compliance from 519 patients who had completed active periodontal treatment up to 7 years earlier and who had begun PM. The patients were classified according to age, sex, diagnosis, prognosis, procedures performed, and compliance with prescribed maintenance intervals. In total, the overall rate of complete compliance was 3.3%, while 57.6% were erratic compliers, and 39.1% never returned for therapy. Female patients complied better than males. Patients who had received surgery complied better with PM than patients who had received scaling and root planing. These results confirmed that the degree of compliance following periodontal therapy is far from ideal. Although universal findings of poor compliance have been found, there may not be a universal solution to resolve the problem. PMID- 12731605 TI - The relationship between periodontal status and peripheral levels of neutrophils in two consanguineous siblings with severe congenital neutropenia: case reports. AB - Congenital neutropenia is characterized by a severe reduction in absolute neutrophil counts, resulting in an almost total absence of neutrophils. It is well known that severe neutropenia affects periodontal status. Oral manifestations include ulcerations, gingival desquamation, gingival inflammation, attachment loss, and alveolar bone loss which may result in tooth loss. Treatment with granulocyte-colony stimulating factor (G-CSF) may improve this periodontal condition. This article reports the relationship between periodontal disease status and peripheral neutrophil levels in two consanguineous siblings with severe congenital neutropenia who did not receive routine G-CSF for 2 years prior to examination. Both siblings were given scaling, root planing, and periodontal prophylaxis in regular follow-up visits. This report demonstrates that periodontal therapy supported by adequate oral hygiene may result in restoration of neutrophil counts in siblings with congenital neutropenia. PMID- 12731606 TI - The predisposing factors of pericoronitis of mandibular third molars in a Jordanian population. AB - OBJECTIVES: The purpose of this study was to identify the most frequently encountered predisposing factors in relation to the etiology of pericoronitis in young adults and to compare these findings with similar studies. METHOD AND MATERIALS: The patients included in this prospective study were those presenting for treatment of signs and symptoms of pericoronitis in the mandibular third molar area during an 8-year period from 1994 to 2001. A standard check sheet of subjective and objective observations was completed, and female patients were asked about menstruation or pregnancy. Each patient with a diagnosis of pericoronitis was interviewed and observations were recorded. Patients were asked about the symptoms relating to the pericoronitis. Patients were divided into five 5-year age groups ranging from 16 to 40 years. RESULTS: During the 8-year period, 2,151 patients presented diagnosed cases of pericoronitis; 932 (43.3%) patients were male and 1,219 (56.7%) were female. The peak age of occurrence of pericoronitis varied from 21 to 25 years (55.2% of the patients). The incidence of pericoronitis was highest in September (207 [9.6%]), followed by April (181 [8.4%]); for subacute pericoronitis, the highest was in February (128 [5.95%]) and April (112 [5.2%]), followed by October (97 [4.5%]). The most frequently seen predisposing factor was upper respiratory tract infection in 815 (37.9%) patients, followed by stress in 473 (22%) patients. CONCLUSION: It was concluded that the upper respiratory tract infection was the most frequently predisposing factor, which could precipitate pericoronitis. PMID- 12731608 TI - Excellence is its own reward. PMID- 12731609 TI - The use of enamel matrix protein in the treatment of localized aggressive periodontitis: a case report. AB - The previously named localized juvenile periodontitis (LJP), recently reclassified as localized aggressive periodontitis (LAP) seen in young people, is a rare condition characterized by severe periodontal destruction around first molars and incisors in individuals with little or no accumulation of visible plaque and/or calculus. Treatment of this condition has traditionally included periodontal surgery and adjunctive antibiotic therapy. Even though several periodontal regeneration techniques in these patients have been reported, there have been no reports of the use of the enamel matrix proteins in the treatment of intrabony defects caused by this disease. This report describes the case of a 15 year-old patient who was diagnosed as having LAP and the resulting treatment and outcome. The treatment consisted of systemic antibiotic therapy and periodontal surgical procedures combined with enamel matrix protein application. The 6-month clinical probing and attachment level measurements and radiographic findings are reported. PMID- 12731607 TI - Branchial cleft anomalies (branchial cleft cyst, cervical lympho-epithelial cyst). PMID- 12731610 TI - Guided bone regeneration using an absorbable membrane combined with a one-stage implant into a recent extraction site: a case report. AB - Guided bone regeneration using an absorbable membrane combined with one-stage implants was used to treat a 45-year-old woman. Two months following tooth extraction, two implants were placed--one in the extraction site and one in a healed site. An absorbable membrane was applied to cover the bony defect in this recent extraction site. After a 24-month follow-up, probing depths were normal, increased radiopacity was seen in the extraction site, and the implant was still clinically stable. PMID- 12731611 TI - Techniques for immediate core buildup of endodontically treated teeth. AB - Access to the pulp chamber for endodontic treatment is indicated inter alia as a result of extensive caries, trauma to the tooth causing fracture or loss of vitality, requiring restoration of the missing tooth structure. Different approaches and materials are described in the literature for foundation restorations, either with a cast post and core or immediately, with a chairside post-and-core system. This article briefly reviews the current data regarding the microbiologic, prosthetic, mechanical, and periodontal aspects while emphasizing the immediate approach using amalgam, resin composites, and glass ionomers. Factors affecting retention of the post are presented to guide the clinician in selecting a suitable post-and-core system to preserve optimal root structure and prevent root fracture. Three clinical cases are presented in which tooth structure was restored using different techniques: in the first two, provisional acrylic resin shells, one custom made and the other prefabricated, were used to house an amalgam coronal-radicular dowel core, where in the third case, a copper band was used for a composite post-and-core system. All cases emphasize the ease of production and short chairtime in the stages of crown fabrication. PMID- 12731612 TI - A comparative clinical study on the Carisolv caries removal method. AB - OBJECTIVE: Carisolv is a relatively new chemomechanical method for caries removal. The aim of this clinical study was to compare Carisolv with the conventional drilling technique. METHOD AND MATERIALS: Forty-five volunteers, ages 18 to 55 years, each with two contralateral primary coronal mesio-occlusal or disto-occlusal carious lesions, similar in extent, participated in the study. Two calibrated operators treated all lesions. One operator treated both lesions in one visit (one lesion with Carisolv and the other with conventional drilling). Following the filling procedure, the opinion of each patient regarding each caries removal method was recorded. The need for drilling in addition to the Carisolv application, the time required for caries removal, the need for anesthesia, and the gingival reaction to the Carisolv gel were recorded by each operator for each case. RESULTS: The patients found Carisolv treatment more pleasant (82%) and preferable (88%) to drilling. Of the patients treated with drilling, 40% required anesthesia compared to the 8% treated with Carisolv. Additional drilling for complete caries removal was needed in 10% of Carisolv treated lesions. Carisolv induced no gingival reaction. Significantly longer times were required for caries removal with Carisolv (12.2 +/- 4.1 minutes) compared to drilling (6.8 +/- 2.8 minutes). CONCLUSIONS: The Carisolv technique was: (a) accepted by the majority of patients, (b) efficient for caries removal, (c) considered a time-consuming technique for the dentists, and (d) considered less dependent on local anesthesia. PMID- 12731613 TI - Dens evaginatus of anterior teeth (talon cusp): report of five cases. AB - The talon cusp, or dens evaginatus of anterior teeth, is a relatively rare dental developmental anomaly characterized by the presence of an accessory cusplike structure projecting from the cingulum area or cementoenamel junction. This occurs in either maxillary or mandibular anterior teeth in both the primary and permanent dentition. This article reports five cases of talon cusp, two of them bilateral, affecting permanent maxillary central and lateral incisors and canines that caused clinical problems related to caries or occlusal interferences. PMID- 12731614 TI - Accidental displacement of a mandibular third molar: a case report. AB - This report describes the rare case of an impacted mandibular third molar that was displaced into the submandibular region. Upon locating the tooth, successful extraction was performed. Healing was unincidental. PMID- 12731615 TI - Implant dentistry for geriatric patients: a review of the literature. AB - The objective of this literature review is to discuss the various parameters related to clinical success of implant treatment in geriatric patients. Old age is not a contraindication for implant therapy; however, clinicians should be aware of potential risks, possible medical complications, and psychosocial issues that affect implant prognosis in geriatric patients. PMID- 12731616 TI - Unusual in vivo extensive corrosion of a low-silver amalgam restoration involving galvanic coupling: a case report. AB - The basic mechanism of dental amalgam corrosion has been thoroughly studied during the last 50 years by various experimental techniques, most often carried out in vitro. Electrochemical methods were extensively employed, and it was recognized that a gradual dealloying of the more electroactive components, Zn, Sn, and to a lesser extent Cu, contributed to change the surface composition. It is also well known that, in all circumstances, galvanic coupling threatens the longevity of the restoration. Among the patients examined at the hospital attached to the Department of Dental Surgery of the Paris 5 University, the authors observed, in a few exceptional circumstances of defective restorations, a release of liquid metal droplets in the surrounding tissue. These particles were identified as elemental mercury. This unexpected phenomenon was the result of the use of a low-silver-content alloy, combined with an enhanced corrosion process due to galvanic coupling with a silver alloy crown and brass screw-posts. In the described clinical case, the tooth, which had to be extracted, was axially sliced for observation by scanning electron microscopy coupled with elemental analysis by X-ray fluorescence energy dispersive spectroscopy. The observations showed a dramatic corrosion of the amalgam matrix with the formation of porosity even in the bulk of the material. Superposition of the element mappings proved (1) the electrochemical coupling of the amalgam with the silver-based crown, (2) the decalcification of the dentin, and (3) the formation of tin oxychloride precipitates in the matrix porosities and the interfacial gaps. These experimental results corroborate the corrosion mechanisms described hereupon. They support the current national and international recommendations for the dental amalgam formulation and for a rigorous professional practice. PMID- 12731617 TI - The microleakage between alloy and polymeric materials in veneer crowns. AB - OBJECTIVE: This study investigated the cervical microleakage between a cobalt chromium (Co-Cr) alloy and four indirect polymeric veneering materials using different adhesive systems. METHOD AND MATERIALS: The Co-Cr-based alloy for fixed prosthesis used in this study (Biosil F, Degussa) was veneered with four different polymeric materials: Artglass, Chromasit, Solidex, and Superacryl. RESULTS: Superacryl produced the highest microleakage score (4 +/- 0.0). The least microleakage at the interface was produced by Artglass (1 +/- 0.816), followed by Chromasit (2 +/- 1.414) and Solidex (2.5 +/- 1.0). Only the differences between Artglass and Superacryl were significant. CONCLUSION: Within the limitations of this in vitro study, cervical microleakage between the coping and the veneer depended on the particular polymeric material used for veneering. Leakage was significantly lower when resin materials were used together with a chemical conditioning system (ie, Artglass/Siloc). PMID- 12731618 TI - Fracture resistance of teeth restored with two different post-and-core designs cemented with two different cements: an in vitro study. Part I. AB - OBJECTIVE: The purpose of this in vitro study was to investigate fracture resistance in teeth restored with cast post and cores with and without ferrule and using two different luting cements. METHOD AND MATERIALS: Forty intact maxillary premolars were endodontically treated after their crowns were removed at 2.00 mm from the cementoenamel junction. Specimens were embedded in acrylic resin blocks in aluminum cylinders, 4.00 mm apical to the cementoenamel junction. Twenty specimens were ferruled, and half of the posts and cores were cemented with zinc-phosphate cement, while the other half was cemented with resin cement. The same procedures were followed for the nonferruled group. Loads were applied at an angle of 45 degrees and measured with a universal testing machine. ANOVA and Tukey test were used for statistical analyses; a significance level was established at 5%. RESULTS: Ferruled specimens showed greater resistance than nonferruled ones, regardless of the cement used. There was no statistical difference between the group of specimens cemented with resin cement and without ferrule and the ferruled groups. The non-ferruled group with zinc-phosphate cement showed the poorest results. CONCLUSION: A 2.00-mm cervical ferrule is important for fracture resistance of restored teeth, and resin cement has a better performance. PMID- 12731619 TI - Freeman-Sheldon syndrome: a case report. AB - Freeman-Sheldon syndrome, also called "whistling-face syndrome," is a very rare genetic condition, occurring both sporadically and by transmission through autosomal dominant or recessive mode, which affects primarily the face and skeleton. Characteristics include microstomia of the mouth, which gives the person a whistling appearance, a flat face, club feet, contracted joint muscles of the fingers and hands, and underdeveloped nose cartilage. This article describes a case of Freeman-Sheldon syndrome in a 10-year-old male. The most apparent deformities are associated with orofacial and skeletal development. PMID- 12731620 TI - Ameloblastoma of the follicular, plexiform, and acanthomatous type in the maxillary sinus: a case report. AB - The ameloblastoma is a benign odontogenic tumor of epithelial origin. Histopathologically, it occurs in six patterns: plexiform, follicular, acanthomatous, granular cell, basal cell, and desmoplastic type. This report presents a rare case of an ameloblastoma in the maxillary sinus that consists of three types: follicular, plexiform, and acanthomatous. The diagnostic procedure, surgical therapy, and follow-up findings are described and discussed. PMID- 12731621 TI - Sialolithiasis. PMID- 12731622 TI - Central facilitation of the glottic closure reflex in humans. AB - The sphincteric function of the larynx, essential to lower airway protection, is most efficiently achieved through strong reflex adduction by both vocal cords. We hypothesize that central facilitation is an essential component of a bilateral adductor reflex and that its disturbance could result in weakened sphincteric closure. Five patients during supraglottic laryngectomy underwent evoked response laryngeal electromyography under 0.5 and 1.0 minimal alveolar concentration (MAC) isoflurane anesthesia. The internal branch of the superior laryngeal nerve was stimulated through bipolar platinum-iridium electrodes, and recording electrodes were positioned in the ipsilateral and contralateral thyroarytenoid muscles. Consistent threshold responses were obtained ipsilaterally from 0.5 to 1.0 MAC anesthesia. However, the contralateral reflex responses approached 0% in successive trials as anesthetic levels approached 1.0 MAC. In human subjects, alteration of central facilitation by deepening anesthesia abolishes the crossed adductor reflex, predisposing to a weakened glottic closure response. A precise understanding of this effect may improve the prevention of aspiration in patients emerging from prolonged sedation or under heavy psychotropic control. PMID- 12731623 TI - Molecular transformation of recurrent respiratory papillomatosis: viral typing and p53 overexpression. AB - Recurrent respiratory papillomatosis (RRP) is a histologically benign disease of the larynx, trachea, and bronchi. Here we report on the histologic and molecular characteristics of 7 cases of malignant transformation of RRP to squamous cell carcinoma (SCCA). The clinical histories of 7 patients with RRP who developed SCCA were carefully reviewed. Sequential biopsies were available from 5 of the 7 cases of spontaneous transformation of RRP to SCCA and were reviewed. In addition, p53 protein overexpression and human papillomavirus (HPV) typing for all cases was examined. The average age of patients with juvenile-onset RRP was 3 years, and that of patients with adult-onset RRP was 31 years. The average age of onset of transformation to SCCA was 28 years. All patients had laryngeal involvement with RRP, and 3 of the 7 patients had tracheal extension of disease. Five patients were tracheotomy-dependent. Four of the 7 patients developed SCCA of the lung, while 3 patients developed laryngeal SCCA. There was no consistent histologic progression from squamous papilloma to papilloma with dysplasia, and all but 1 of the SCCAs were well differentiated. The overexpression of p53 protein was variable in each of the 5 patients. We detected HPV types 6/11 in papillomas from 3 patients, and HPV types 6/11, 16/18, and 31/33/51 in a papilloma of a fourth patient. No HPV DNA was detected in papillomas of 2 patients. We found HPV 6/11 in 4 of the carcinomas. We conclude that the spontaneous transformation of RRP to SCCA is not characterized by a histologic progression through dysplasia over time. Transformation can result in the loss of HPV expression. It does not appear that p53 is a molecular marker for monitoring the transformation process. Thus, these cancers may be very difficult to diagnose histologically and clinically early in the course of the transformation of the disease. PMID- 12731624 TI - Posterior cricoarytenoid myoplasty with medialization thyroplasty in the management of refractory abductor spasmodic dysphonia. AB - Of the approximately 100,000 Americans with primary (idiopathic) laryngeal dystonia, 10% to 15% are thought to havethe abductor form. Botulinum A toxin injected into the posterior cricoarytenoid muscle and/or cricothyroid muscle has been employed as the "gold standard" for therapeutic management; however, successful results are significantly less frequent than with injections for the adductor form. This report describes a new phonosurgical procedure, posterior cricoarytenoid myoplasty with medialization thyroplasty, designed for these refractory patients. Posterior cricoarytenoid myoplasty with medialization thyroplasty has been performed on 3 patients with abductor laryngeal dystonia. All patients had failed at least 5 previous botulinum A injections to the posterior cricoarytenoid and cricothyroid muscles. All patients underwent preoperative and 3 postoperative (2 weeks, 3 months, and 1 year) phonatory analyses. Analysis consisted of recording an aloud reading of a standard passage while a blinded trained speech pathologist counted prolonged voiceless consonants. The patients also completed a satisfaction survey at 1 year. The results demonstrated significant, long-lasting, uniform reduction in breathy breaks in all subjects. The participants all judged their symptoms as greatly improved. Bilateral procedures may be necessary, but should be staged to prevent possible airway compromise. When applied appropriately, posterior cricoarytenoid myoplasty with medialization thyroplasty is a viable tool in the management of refractory abductor laryngeal dystonia. PMID- 12731625 TI - Microvascular transplantation of tracheal allografts model in the canine. AB - The inability to reconstruct extensive and often life-threatening tracheal defects is a clinical dilemma. The objective of this study was to achieve microvascular revascularization and transplantation of long-segment circumferential tracheal allografts in a canine model. Fifteen mongrel dogs were randomly assigned to 5 treatment groups. Twelve dogs underwent an excision of an 8-cm tracheal segment followed by transplantation and microvascular revascularization of an 8-cm cervical trachea allograft. Group 1 (n = 4) was treated with 10 mg/kg per day of cyclosporin A (CsA) and 7.5 mg/kg per day of mycophenolate mofetil (MM). Group 2 (n = 4) was treated with 5 mg/kg per day of CsA and 7.5 mg/kg per day of MM. Group 3 (n = 4) was treated with 2.5 mg/kg per day of CsA and 7.5 mg/kg per day of MM. Group 4 (n = 2) underwent an autograft tracheal transplant and received postoperative 2.5 mg/kg perday of CsA and 7.5 mg/kg per day of MM. Group 5 (n = 1) did not undergo surgery, but received postoperative 2.5 mg/kg per day of CsA and 7.5 mg/kg per day of MM. The animals were maintained for a duration of 30 days, during which time the graft was assessed by routine endoscopic examination and tracheal biopsies. Ex vivo, tracheal autografts were examined grossly for graft healing and microscopically for histologic architecture. The mean survival times were 13.25 days (group 1), 16 days (group 2), and 20 days (group 3). There was 1 early allograft failure secondary to microvascular thrombosis, and there were 4 delayed failures secondary to postoperative wound infections. Five dogs were euthanized before the end of the 30-day observation period because of failure to thrive or hypocalcemic tetany. None of the dogs in the study demonstrated endoscopic or histologic evidence of rejection before euthanasia. Postmortem examination of the surviving dogs demonstrated normal histologic architecture without evidence of rejection. For the first time, we have achieved allotransplantation of long tracheal segments based on the cranial thyroid artery and internal jugular vein. Minimal systemic immunosuppression appears to be associated with a higher survival rate and a lower complication rate. PMID- 12731626 TI - Electromyographic findings in recurrent laryngeal nerve reinnervation. AB - Abductor, adductor, and combined reinnervation procedures have been explored with variable success rates. We describe the experience of a tertiary care center with adductor reinnervation procedures, including preoperative and postoperative videostroboscopy and electromyography (EMG) findings. A retrospective chart review was performed from 1997 to 2001 that included 9 patients. Preoperative and postoperative voice comparison was performed by 3 blinded speech pathologists. Clinical comparisons of videostroboscopy findings for vocal fold bulk, tone, position, presence of gap, and movement are elucidated. The preoperative and postoperative EMG findings are described. In all patients, preoperative EMG revealed a dense, complete denervation of the affected recurrent laryngeal nerve. No movement was noted on videostroboscopy with persistent glottic gap. Reinnervation involved a nerve-muscle pedicle or a direct neurorrhaphy of the ansa cervicalis to the recurrent laryngeal nerve. Voice improvement was noted between 60 days and 3 months after reinnervation. Four postoperative EMG studies were performed. An early postoperative EMG study at 5 months revealed activation of the lateral cricoarytenoid muscle and thyroarytenoid muscle with head-lift. Videostroboscopy showed excellent near-midline static positioning of the vocal fold. Late EMG studies, performed 12 to 16 months after reinnervation, revealed "learning" of these muscles, with new activation on "eee" phonation. We conclude that recurrent laryngeal nerve reinnervation procedures belong in the armamentarium of the laryngologist for the treatment of vocal fold paralysis. The EMG findings reported in this study suggest that ongoing reinnervation allows for activation with phonation in matured neuronal anastomoses. Overall, this procedure results in excellent patient acceptance and near-normal vocal quality. PMID- 12731627 TI - Cepstral peak prominence: a more reliable measure of dysphonia. AB - Quantification of perceptual voice characteristics allows the assessment of voice changes. Acoustic measures of jitter, shimmer, and noise-to-harmonic ratio (NHR) are often unreliable. Measures of cepstral peak prominence (CPP) may be more reliable predictors of dysphonia. Trained listeners analyzed voice samples from 281 patients. The NHR, amplitude perturbation quotient, smoothed pitch perturbation quotient, percent jitter, and CPP were obtained from sustained vowel phonation, and the CPP was obtained from running speech. For the first time, normal and abnormal values of CPP were defined, and they were compared with other acoustic measures used to predict dysphonia. The CPP for running speech is a good predictor and a more reliable measure of dysphonia than are acoustic measures of jitter, shimmer, and NHR. PMID- 12731628 TI - Histology of nerves and muscles in adductor spasmodic dysphonia. AB - To elucidate the etiology and pathophysiology of spasmodic dysphonia, we examined the adductor branch of the recurrent laryngeal nerve and the lateral cricoarytenoid muscle from 9 consecutive patients with this disorder who were previously treated with botulinum toxin. Histologic examination revealed average muscle fiber diameters ranging from 21 to 57 microm. Botulinum toxin treatment related muscle atrophy was observed up to 5 months after injection. Endomysial fibrosis was present in all samples. Histochemical analysis in 8 patients revealed type 2 fiber predominance in 7 patients and fiber type grouping in 2. Type-specific muscle fiber size changes were not present. Nerve samples were examined in plastic sections. In 8 patients the nerves contained homogeneous, large-diameter myelinated nerve fibers and sparse small fibers. One patient had a relatively increased proportion of small myelinated nerve fibers. Overall, the nerve fiber diameter was slightly larger in patients than in controls. These findings may implicate the central nervous system in the pathophysiology of adductor spasmodic dysphonia. PMID- 12731629 TI - Is secretory otitis media a single disease entity? AB - The outcome of 809 children who had middle ear effusion (MEE) was correlated with their age and medical history and the bacteriologic and cytologic findings of the MEE. Three groups emerged. Group A (n = 384) had a medical history of relatively recent acute otitis media (AOM) with a peak prevalence of 2.6 years of age, and its MEE was hypercellular (mostly polymorphonuclear leukocytes) with a 24% rate of positive bacteriologic culture. In contrast, the peak prevalence of group B (n = 280) was approximately 5 years of age. Its MEE followed no otologic medical history and was bacteriologically sterile with relatively few cells (mostly lymphocytes typical of viral infections). Group AB (n = 145) resembled group B except that they had a history of AOM some years before hearing loss onset. Altogether, the clinical features of group A are statistically distinguishable from those of groups B and AB in most respects. These findings explain the bimodal peak prevalence distribution that was found in many available epidemiological studies of secretory otitis media (SOM) and that was also seen in our data. It appears that SOM should neither be termed nor treated as an otitis media, but as a sequela of either bacterial AOM (group A) or of insidious, asymptomatic, probably viral otitis media (groups B and AB). Our data do not support antibiotic treatment for SOM (otitis media with effusion)--especially not in chronic cases. PMID- 12731630 TI - Fixation of the anterior mallear ligament: diagnosis and consequences for hearing results in stapes surgery. AB - In the search for possible causes of unfavorable results after stapes surgery, the study reported here focused on the anterior mallear ligament, since it has been previously reported that partial mallear fixation (PMF) leads to functional failure in 38% of cases of stapes revision surgery. The aims of the study were to identify effective methods for the diagnosis of PMF and experimentally assess the conductive hearing loss that results from PMF. The study included vibration amplitude measurements of the ossicles by laser Doppler interferometry (LDI) in 19 patients and 5 fresh human temporal bone (TB) specimens. Analysis of their dynamic behavior was performed by finite element modeling (FEM). Similar, significant changes of manubrium vibration patterns for PMF were found by FEM calculations, in TB experiments, and in patients. We could identify PMF either before operation, using LDI, or during operation, by manual palpation. In the TB experiments and FEM calculations, the attenuation of the stapes displacement due to an isolated PMF was approximately 10 dB and frequency-dependent. Untreated anterior mallear ligament fixation produced a persistent air-bone gap of approximately 10 dB after stapedioplasty. PMID- 12731631 TI - Neuroepithelial cysts of the middle ear. AB - Neuroepithelial cysts are lesions of the central nervous system that have previously been reported in cerebral parenchyma, in the spinal cord, and within the ventricles in association with the choroid plexus. We describe 2 cases of neuroepithelial cysts of the middle ear. One was diagnosed after surgery for a retraction pocket and chronic otitis media complicated by an intraoperative cerebrospinal fluid (CSF) leak. The other produced bilateral spontaneous CSF otorrhea and mimicked the presentation of arachnoid granulations. Both cases were verified with immunohistochemical stains. Neuroepithelial cysts, although rare, should be considered in the differential diagnosis of spontaneous CSF otorrhea or an epitympanic mass. PMID- 12731632 TI - Laryngocele resection by combined external and endoscopic laser approach. AB - Options in the management of laryngoceles include observation, endoscopic resection, and resection via an external approach. We introduce a combined endoscopic and external approach that we have employed on several occasions to ensure complete removal of the laryngocele and the saccule from which it originated. A case is presented to help define the technique. PMID- 12731633 TI - Update on treatment of Marshall's syndrome (PFAPA syndrome): report of five cases with review of the literature. AB - Marshall's syndrome or PFAPA (periodic fever, aphthous stomatitis, pharyngitis, cervical adenitis) syndrome is a recently described pediatric periodic disease characterized by recurrent febrile episodes associated with head and neck symptoms. The origin of this syndrome is unknown, and it can last for several years. During healthy periods, patients grow normally. The differential diagnosis includes other diseases characterized by periodic fevers, such as familial Mediterranean fever, familial Hibernian fever, hyperglobulinemia D syndrome, Behcet's disease, cyclic neutropenia, juvenile rheumatoid arthritis, and several infectious diseases. Many treatments have been used, with various results, including antibiotics, nonsteroidal anti-inflammatory drugs, acetylsalicylic acid, colchicine, antiviral medicines, steroids, cimetidine, and tonsillectomy. We describe 5 new patients affected by PFAPA syndrome who were observed at the Department of Pediatric Otorhinolaryngology, Spedali Civili, Brescia, Italy, from November 2000 to August 2001. All children underwent physical examination, bacterial, fungal, and viral cultures, chest radiography, and several laboratory studies. The patients were treated by successful tonsillectomy, and after a mean follow-up of 10 months, no recurrence was observed. An analysis of the literature is also presented with particular emphasis on the differential diagnosis of this rare illness and the results of the different therapeutic options. PMID- 12731634 TI - Isolated inflammatory sphenoid sinus disease in children. AB - Isolated inflammatory disease of the sphenoid sinus is very uncommon in the pediatric population. A 10-year review of all patients at our institution 16 years of age or younger with inflammatory sphenoid sinus disease found 8 patients with isolated sphenoid sinusitis and 3 with sphenoid mucoceles. The most common symptoms were headache and visual disturbance. Five patients with uncomplicated sinusitis were successfully managed medically, while 3 with either complicated sinusitis or sinusitis not responding to antibiotics were treated by endoscopic sphenoidotomy. All patients with a mucocele were treated surgically. Isolated inflammatory sphenoid sinusitis should be considered in children age 7 years or older who present with headache that does not respond to simple analgesia. Delayed diagnosis and advanced disease may lead to life-threatening complications. PMID- 12731635 TI - Inhalational anesthetic technique in microlaryngeal surgery: a comparison between sevoflurane-remifentanil and sevoflurane-alfentanil anesthesia. AB - We studied the effects of sevoflurane, remifentanil hydrochloride, and alfentanil anesthesia in terms of the hemodynamic responses and emergence characteristics of patients scheduled for elective microlaryngeal surgery. Sixty patients (ASA I to III) were randomly allocated into 2 groups: group S-R (sevoflurane-remifentanil) and group S-A (sevoflurane-alfentanil; 1:20 and 1:4 ratios of remifentanil to alfentanil for induction and maintenance of anesthesia, respectively; doses not strictly equipotent). The mean arterial pressure and heart rate were measured before and after induction of anesthesia, 1 and 3 minutes after endotracheal intubation, at the insertion of the operating laryngoscope, and every 3 minutes during surgery. The emergence times and side effects during the first 30 minutes after surgery were also recorded. The mean arterial pressure values at the insertion of the operating laryngoscope and throughout the procedure were significantly greater (p < .05) in group S-A than in group S-R. The emergence times and postoperative side effects did not differ, except for the greater pain score (p < .05) in group S-R. In conclusion, sevoflurane with remifentanil seems to maintain cardiovascular stability during microlaryngeal surgery more effectively than sevoflurane with alfentanil. Both anesthetic regimens seem to provide rapid and uneventful emergence. PMID- 12731636 TI - Hazardous foreign bodies: complications and management of button batteries in nose. AB - Miniature batteries are easily available in our domestic environment, powering many electronic devices and toys. Despite improvement in the safety standards, children are able to remove the batteries from these devices. These batteries pose a hazard to children, as they are small and easily inserted into the nose or ears or even swallowed. We describe 6 children who inserted button batteries into their noses. Four of these insertions resulted in septal perforations. The mechanisms and management of button battery injury are discussed. We emphasize the need for urgent removal of a battery from the nose to prevent long-term complications. PMID- 12731638 TI - Obstructive congenital gingival granular cell tumor. AB - Congenital gingival granular cell tumors (also known as congenital epulis or Neumann's tumor) are rare and always benign intraoral tumors originating from the alveolar ridge. They are typically seen as a mass protruding out of a newborn child's mouth. We report a case of a large obstructive congenital gingival granular cell tumor of the mandibular ridge. The intraoral mass was first detected on a 38-week prenatal ultrasound scan and resulted in neonatal airway obstruction. Complete surgical removal was performed with an uneventful postoperative course. Histologic and immunohistochemical assessments with antibodies against S-100 protein confirmed the diagnosis. The clinical and morphological aspects, differential diagnosis, histogenesis, and treatment are discussed. PMID- 12731637 TI - Prevalence of otomycosis in outpatient department of otolaryngology in Tribhuvan University Teaching Hospital, Kathmandu, Nepal. AB - Otomycosis is a not-uncommon clinical problem encountered in our otolaryngology practice. We report 100 diagnoses (107 ears) of otomycosis, of which, on microbiological examination, 87 specimens (81.3%) showed positive fungal cultures. Of these, a single isolate was found in 85 cases (79.4%), mixed isolates were found in 2 cases, and 20 cases (18.7%) had no growth. The most common fungal pathogen found was Aspergillus, followed by Candida albicans. The causative factors for otomycosis were examined. The results of treatment with clotrimazole were analyzed. PMID- 12731639 TI - Fetal and neonatal neurologic case histories: assessment of brain disorders in the context of fetal-maternal-placental disease. Part 2: Neonatal neurologic consultations in the context of adverse antepartum and intrapartum events. AB - The more conventional role of the pediatric neurologist involves the evaluation of the child after birth. Although the pediatric neurologist rarely attends the delivery of the neonate, consultation by the neurologist should begin immediately following stabilization by the neonatal resuscitation team. Four interrelated aspects of the neurologist's clinical assessment will be discussed in the context of reaching a consultative opinion, which must incorporate knowledge of chronologic events before as well as during labor and delivery. This evaluation encompasses an assessment of levels of arousal, increased or decreased muscle tone, presence of seizures, and effects of systemic diseases on the central nervous system, which are the essential elements of a complete neurologic examination. Documentation of the neonate's neurologic condition, together with knowledge of maternal, fetal, and placental diseases, will help anticipate neuroresuscitative decisions, as well as subsequent neurologic deficits. PMID- 12731640 TI - Prospective preliminary analysis of the development of autism and epilepsy in children with infantile spasms. AB - The objective of this study was to compare the efficacy of corticotropin (ACTH) versus vigabatrin in treating infantile spasms and to determine which medication has a more favorable long-term outcome in terms of cognitive function, evolution of epilepsy, and incidence of autism. Patients with infantile spasms were included in the study if they were 3 to 16 months old, had hypsarrhythmia, and had no previous treatment with vigabatrin or corticosteroids. Patient evaluation included electroencephalographic and psychometric measures before and after treatment. Patients were stratified based on etiology (idiopathic or symptomatic) and sex and then randomized between the ACTH and vigabatrin treatment groups. Each of the treatment groups received either ACTH or vigabatrin for 2 weeks. At the end of 2 weeks of treatment, patients were considered responders if spasms and hypsarrhythmia resolved. Nonresponders were crossed over and treated with the alternate drug. Nine patients were included in the study. Three patients received ACTH, one of whom was a responder. Six patients received vigabatrin, three of whom were responders. The five nonresponders received both therapies. All patients had some degree of developmental plateau or regression before the initiation of treatment. Four patients with idiopathic infantile spasms showed improved cognitive function following treatment. The remaining five patients remained significantly delayed. Five patients with symptomatic infantile spasms had epilepsy following treatment; three of them were in the autistic spectrum. The small number of infants in this pilot study is insufficient to determine which of the two drugs is more effective. However, the following trends were identified: vigabatrin may be more effective for patients with symptomatic infantile spasms; patients with idiopathic infantile spasms tend to have a better cognitive outcome; and patients with symptomatic infantile spasms tend to develop both epilepsy and autism. PMID- 12731641 TI - Hypotonia, congenital hearing loss, and hypoactive labyrinths. AB - The records of all patients attending a neurosensory genetics clinic over an 11 year period were reviewed. Of the 450 patients seen, 31 presented with sensorineural hearing loss, hypotonia, and delay in the acquisition of motor milestones. Of these, 4 children were found who did not have an etiologic diagnosis such as Down syndrome or cerebral palsy. Vestibular testing revealed hypoactive labyrinthine function in all 4 of the cases, and careful imaging of the temporal bone showed anomalous development of the cochlea, vestibule, and semicircular canals in 3 of the 4 cases. None of the patients had ataxia, tremor, or significant nystagmus. Over time, the hypotonia improved in all, and none were felt to have cognitive deficits. These cases demonstrate that hypoactive labyrinthine function may be associated with hypotonia that is severe enough to result in delayed acquisition of motor milestones. The patients followed the typical remitting course of "benign congenital hypotonia." The distinguishing clinical feature is the presence of moderate to profound sensorineural hearing loss in all of the patients. PMID- 12731642 TI - Epilepsy in children with cerebral palsy. AB - To study the spectrum of epilepsy in children with cerebral palsy, 105 consecutive children with cerebral palsy and active epilepsy, between 1 and 14 years of age, were studied prospectively. A detailed history and examination, electroencephalography (EEG), and computed tomography (CT) were done in all cases. The social quotient was assessed using the Vineland Social Maturity Scale. A retrospective cohort of 452 cases of cerebral palsy was studied to find the prevalence of epilepsy in cerebral palsy. A control group of 60 age-matched children with cerebral palsy but no epilepsy was also studied for comparison of the social quotient. Of the 105 children, 65 were male, 40 of 105 (38%) had a history of birth asphyxia. The mean age of onset of seizures was 18.9 months; 64 (60.95%) had seizure onset before 1 year of age. Children with myoclonic seizures (P < .05) and infantile spasms (P < .01) had seizure onset significantly early in life. Generalized seizures were the most common, followed by partial seizures, infantile spasms, and other myoclonic seizures. Seizures were controlled in 45 (58.1%) children, and polytherapy was required in 40 children. EEG and CT abnormalities were seen in 70.5% and 61% of the children. Seizure control was achieved in 74% of the patients with a normal to borderline social quotient compared with 48.7% with a social quotient less than 70. Social quotient values had a positive correlation with age of onset of seizures (P < .01) and with better control of seizures (P < .01). Of the cohort of 452 children, 160 (35.4%) had epilepsy. The maximum incidence (66%) was seen in children with spastic hemiplegia, followed by quadriplegia (42.6%) and diplegia (15.8%). Epilepsy in cerebral palsy is seen in about one third of cases; it is often severe and difficult to control particularly in children with mental retardation. PMID- 12731643 TI - Pediatric neurology services in Canada: demand versus supply. AB - As the practice of pediatric neurology has continued to evolve and expand, concerns have emerged regarding a perceived trend toward increasing clinical demand and decreasing manpower. Quantitative data are lacking, however. To address this, a questionnaire was sent to hospital-based pediatric neurology units in Canada, with all 18 centers responding. A total of 63 full-time equivalent pediatric neurologists were disproportionately spread across the country, giving an overall ratio of 2.1 per million population (or 1.1 per 100,000 children). Waiting times for nonurgent consultations showed a median of 12.5 weeks. The number of weekly clinics per population was not proportional to either the number of specialists per center or to the waiting times. The regional variations in the level of service do not correlate with the regional manpower figures, reflecting different individual profiles of clinical and academic activities. PMID- 12731644 TI - Unfolding story of inclusion-body myositis and myopathies: role of misfolded proteins, amyloid-beta, cholesterol, and aging. AB - Sporadic inclusion-body myositis and hereditary inclusion-body myopathies are progressive muscle diseases leading to severe disability. We briefly summarize their clinical pictures and pathologic diagnostic criteria and discuss the latest advances in illuminating their pathogenic mechanism(s). We emphasize how different etiologies might lead to the strikingly similar pathology and possibly similar pathogenic cascade. On the basis of our research, several processes seem to be important in relation to the still speculative pathogenesis, including (a) increased transcription and accumulation of amyloid-beta precursor protein and accumulation of its proteolytic fragment amyloid-beta; (b) abnormal accumulation of components related to lipid metabolism, for example, cholesterol, accumulation of which is possibly owing to its abnormal trafficking; (c) oxidative stress; (d) accumulations of other Alzheimer's disease-related proteins; and (e) a milieu of muscle cellular aging in which these changes occur. We discuss a potentially very important role of unfolded and/or misfolded proteins as a possible mechanism in the formations of the inclusion bodies and other abnormalities. PMID- 12731645 TI - Cardiac complications of childhood myopathies. AB - The management of individuals with a neuromuscular disorder is usually focused on the skeletal muscle weakness and resulting complications, such as respiratory failure. Long-term prognosis of a number of neuromuscular conditions is, however, also determined by the type and severity of cardiac involvement. Early recognition and treatment of the cardiovascular complications are part of the task of the multidisciplinary team involved in the care of these patients. Although for several of the common conditions, there is general consensus on the cardiac investigations and treatments, in the rarer disorders, evidence-based recommendations are not available, and suggestions from experts provide an acceptable solution. This review summarizes the recent advances in our understanding of the pathogenesis and phenotypic diversity of cardiac complications associated with pediatric myopathies and provides a rational framework for planning the monitoring and therapeutic intervention in individual conditions. PMID- 12731646 TI - Prevalence of neuromuscular diseases in Chinese children: a study in southern China. AB - Our objective was to study the prevalence of neuromuscular diseases in Chinese children. A prospective study of neuromuscular diseases in Chinese children was conducted from 1985 to 2001 in Hong Kong, which is a city in southern China. The population census of June 30, 2001, was used to calculate the prevalence of neuromuscular diseases in Chinese children. Altogether, 332 children aged < 19 years at first assessment with neuromuscular diseases confirmed by using electromyography, muscle biopsy, and/or molecular genetic study were included in the study. Of these, 228 (68%) had inherited and 104 (32%) had noninherited neuromuscular diseases. Of the inherited neuromuscular diseases, the most common were the dystrophinopathies, including Duchenne muscular dystrophy (n = 66) and Becker muscular dystrophy (n = 8). Spinal muscular atrophy was the second most common (n = 61). Of the noninherited neuromuscular disorders, myasthenia gravis was the most common (n = 62, 60%). Nearly 88% of the cases of myasthenia gravis were ocular type. The prevalence rate of neuromuscular diseases in June 2001 (n = 291 surviving) is estimated to be 214 x 10(-6). The estimated prevalence rate of neuromuscular diseases in our Chinese children is 1 in 4669. PMID- 12731647 TI - Pontocerebellar hypoplasia type 1: new leads for an earlier diagnosis. AB - Pontocerebellar hypoplasia type 1 is a rare disease characterized by pontocerebellar hypoplasia and anterior horn cell degeneration. The oldest reported child died at the age of 26 months. Two siblings were diagnosed with pontocerebellar hypoplasia type 1 after the death of the second sibling at 40 months of age from respiratory failure and the unexpected finding of anterior horn cell degeneration on her autopsy. The older sibling was a boy who was labeled as having cerebral palsy. He died at 14 months of age from pneumonia following a clinical course similar to his sister's, who was born 5 years after his death. Both siblings had significant global developmental delay with axial and peripheral hypotonia initially. Peripheral hypertonia with brisk reflexes developed later but were absent prior to death. Extensive investigations in the second sibling ruled out known metabolic (including congenital disorders of glycosylation) and mitochondrial diseases using skin fibroblast cultures and enzyme analysis. Genetic testing for Friedreich's ataxia; neuropathy, ataxia, and retinitis pigmentosa (NARP); spinal muscular atrophy; and spinocerebellar ataxia type 1, 2, 3, 6, 7, and 8 gene abnormalities was negative. The elecroretinogram showed a previously unreported finding of abnormal and progressive rod/cone response. Our cases provide clinical and previously unreported electroretinographic evidence for neurodegeneration in pontocerebellar hypoplasia type 1 and call for the expansion of the disease phenotype. PMID- 12731648 TI - Rhombencephalosynapsis associated with cutaneous pretibial hemangioma in an infant. AB - Rhombencephalosynapsis is a rare congenital abnormality characterized by noncleavage of the cerebellar hemispheres and dentate nuclei with agenesis or severe hypoplasia of the cerebellar vermis. We report an 8-month-old boy with hypotonia, mental and motor retardation, and capillary hemangiomas on his pretibial region. Cranial magnetic resonance imaging revealed vermian hypoplasia with fused cerebellar hemispheres called rhombencephalosynapsis. Other cerebellar and cerebral structures were normal. With this constellation of findings, we attempt to identify the pathogenesis of this posterior fossa malformation. We also discuss cerebellar embryogenesis to differentiate this condition from other posterior fossa malformations such as Dandy-Walker malformation and Joubert's syndrome. PMID- 12731650 TI - Unilateral schizencephaly and contralateral polymicrogyria associated with umbilical cord mass. AB - We report a 6-month-old boy with diffuse hypertonia and developmental delay who had unilateral separated-lip schizencephaly and contralateral polymicrogyria. The contralateral polymicrogyria was associated with an incomplete clefting in that hemisphere. An umbilical cord hamartoma is presumed to have caused hypoperfusion to the early developing brain, resulting in bilateral lesions. PMID- 12731649 TI - Auditory processing studied prospectively in two hemidecorticectomy patients. AB - Auditory processing of speech and nonspeech sounds was studied prospectively in two hemidecorticectomy patients (ages 10-11 years) with Rasmussen's syndrome. We tested auditory word recognition under four listening conditions: in quiet, in noise, after acoustic filtering, and dichotically. Recognition of environmental sounds and discrimination of tones and digitized syllables were also tested. Presurgical testing confirmed normal processing of speech and nonspeech, for both patients, under all listening conditon. One year after surgery, both patients demonstrated intact recognition of words and environmental sounds in quiet but impaired word recognition in noise. The left hemidecorticectomy patient also demonstrated impaired recognition of low-pass filtered words. These findings suggest that either hemisphere can process speech or nonspeech sounds in quiet, whereas both hemispheres are needed to process speech in background noise. Hemispheric contributions to processing speech in noise appear to differ, with the left hemisphere compensating for loss of phonologic information and the right hemisphere compensating for increased attention demands. PMID- 12731651 TI - Rod distribution and muscle fiber type modification in the progression of nemaline myopathy. AB - Nemaline myopathy is a structural congenital myopathy associated with the presence of rodlike structures inside the muscle fibers and type I predominance. It may be caused by mutations in at least five genes: slow alpha-tropomyosin 3 (chromosome 1q22-23), nebulin (chromosome 2q21.1-q22), actin (chromosome 1q42), tropomyosin 2 (chromosome 9p13), and troponin T1 (chromosome 19q13.4). The effect of these mutations in the expression of the protein and the mechanism of rod formation is still under investigation. We analyzed the possibility of progressive alterations with time and/or disease evolution, such as transformation of type I to type II fiber and rod pattern and distribution in muscle fibers from patients with nemaline myopathy, through a morphometric and immunohistochemical analysis of different muscle protein isoforms. A tendency of diffuse rods to be organized in the subsarcolemmal region was observed in two patients who were submitted to subsequent biopsies after 10 and 13 years. Additionally, we observed the expression of type II protein isoforms in type I fibers and a higher proportion of type II fibers in the younger patient of a pair of affected sibs, giving further support to the hypothesis of progressive conversion of type II to type I fibers in nemaline myopathy. PMID- 12731653 TI - Quantitative analysis of desmethylselegiline, methamphetamine, and amphetamine in hair and plasma from Parkinson patients on long-term selegiline medication. AB - Hair and plasma from patients on long-term selegiline medication were analyzed to evaluate the relationships between plasma and hair melanin concentrations and the incorporation of the selegiline metabolites methamphetamine and amphetamine in hair, and to evaluate hair analyses for determining compliance in medication. Analyses were performed on both the whole hairs, as well as pigmented and non pigmented hairs from gray-haired patients. Melanin was quantitated by spectrophotometry, and metabolites were quantitated by gas chromatography-mass spectrometry. Concentrations in pigmented and non-pigmented hairs differed significantly for both methamphetamine (p < 0.01) and amphetamine (p < 0.02), with mean concentration ratios being 3.69 +/- 1.88 and 2.95 +/- 1.16 for methamphetamine and amphetamine, respectively. Segmental analysis indicated that some patients had not been compliant with medication. We concluded that the incorporation of methamphetamine and amphetamine into hair of single individuals shows a preference for pigmented hairs over white hairs and that segmental analysis of hair may be useful when measuring compliance with medication. PMID- 12731652 TI - Cocaine, benzoylecgonine, amphetamine, and N-acetylamphetamine binding to melanin subtypes. AB - Experiments have been performed to document the in vitro binding of cocaine, benzoylecgonine (BE), amphetamine, and N-acetylamphetamine (N-AcAp) to synthetic melanin subtypes. The two predominant melanin types in hair are the black eumelanins and the reddish-brown pheomelanins. The melanins included in this study are two black eumelanin subtypes [5,6-dihydroxyindole (DHI) and 5,6 dihydroxyindole-2-carboxylic acid (DHICA) derived melanins], a reddish-brown pheomelanin [from 5-cysteinyl-S-Dopa (5-CysDOPA)], and two mixed eu-/pheomelanin copolymers. Results indicate that the basic drugs cocaine and amphetamine bind to eumelanins and mixed eu-/pheomelanins to varying degrees, but not to pure pheomelanin. BE and N-AcAp, net neutral molecules, do not bind to any type of melanin. As a model of which eumelanin chemical functional groups bind drugs, amphetamine was shown, using tandem mass spectrometry, to form a noncovalent adduct with dimerized oxidized catechol. Similar functional groups on the eumelanin polymer may represent an important drug-binding site. Overall, these findings show that basic drugs have a greater affinity for melanin than their net neutral analogues, reveal that melanin types differ when it comes to drug binding, help elucidate what properties of melanin are important for drug binding, and help explain why hair color biases exist. PMID- 12731654 TI - Procainamide and quinidine inhibition of the human hepatic degradation of meperidine in vitro. AB - Procainamide and quinidine inhibition of the degradation of meperidine in human liver was investigated by incubation of two concentrations of either drug with meperidine in homogenates of human liver over 24 and 36 h. Meperidine concentrations declined by 26% after incubation for 24 h and by 42% after incubation for 36 h. In the presence of procainamide, however, they decreased by only 15% to 18% at 24 h and by only 26% to 28% at 36 h. In the presence of quinidine, they declined by only 18% to 19% at 24 h and by only 27% to 28% at 36 h. Procainamide and quinidine may inhibit human hepatic carboxylesterase hCE-1, which is responsible for catalyzing the hydrolysis of meperidine. This inhibition may prolong the biological half-life of meperidine in patients receiving the drug together with either procainamide or quinidine. PMID- 12731655 TI - Distribution of butalbital in postmortem tissues and fluids from non-overdose cases. AB - During the investigation of fatal aviation accidents, postmortem samples from the pilots/co-pilots are submitted to the Federal Aviation Administration's (FAA) Civil Aerospace Medical Institute (CAMI) for toxicological analysis. Although therapeutic levels for most drugs are typically reported in the scientific literature for blood and plasma, blood specimens are received in only approximately 70% of our cases. Therefore, it is imperative for an accident investigator and forensic toxicologist to be able to estimate drug concentrations in an aviation accident victim's blood from available tissue drug concentrations. This is exemplified by a recent aviation fatality in which butalbital was identified in the muscle tissue of a pilot. In this case, no blood was available for analysis, but investigators needed to know the approximate butalbital concentration expected in the victim's blood. Certain side effects of butalbital, such as drowsiness, sedation, dizziness, and a feeling of intoxication, could affect pilot performance and become a significant factor in an aviation accident. Thus, our laboratory determined the distribution of butalbital in various postmortem tissues and fluids. The distribution coefficients for butalbital, expressed as specimen/blood ratios, were found to be as follows: 0.66 +/- 0.09 (muscle, n = 4), 0.98 +/- 0.09 (kidney, n = 4), 0.87 +/- 0.06 (lung, n = 4), 0.75 +/- 0.03 (spleen, n = 4), 0.96 +/- 0.07 (brain, n = 3), 2.22 +/- 0.04 (liver, n = 4), and 0.91 +/- 0.17 (heart, n = 2). The results obtained from our limited number of cases suggest that muscle, kidney, lung, spleen, brain, liver, and heart could be used, in a cautious and conservative fashion, to estimate butalbital blood concentrations. PMID- 12731656 TI - Ofloxacin as a reference marker in hair of various colors. AB - It has been proposed that administration of a reliable marker substance to human subjects may enhance the ability to identify drug use and treatment compliance in drug treatment programs. The goal of this study was to determine if an oral dose of the antibiotic ofloxacin (OFLX) could be used as a "marker" substance to establish reference points with respect to time in hair of various colors. Male and female subjects (n = 32) between 18 and 40 years of age received 800 mg of OFLX as a divided oral dose on a single day. Subjects were restricted from cutting their hair or performing chemical treatments. Hair was collected (by cutting) before, and at weeks 4, 5, 6, and 7 after drug administration. Subjects were classified as having black (n = 5), brown (n = 13), blonde (n = 8), or red (n = 6) hair. Hair was segmented into 3.0-cm segments prior to digestion, extraction, and analysis by high-pressure liquid chromatography (HPLC). At 7 weeks, the mean OLFX concentrations (+/- 1 SD) in the first 3.0 cm of hair closest to the scalp were as follows: 30.6 +/- 8.5 ng/mg (black), 6.0 +/- 1.8 ng/mg (brown), 3.5 +/- 1.6 ng/mg (blonde), and 1.4 +/- 0.3 ng/mg (red). A similar pattern was found in hair collected at weeks 4-6. Quantitative eumelanin (EUM) hair concentrations for each subject were also determined for each subject via HPLC. A strong relationship between OFLX concentration at 7 weeks and EUM was noted (r2 adjusted = 0.728; p < 0.001). In six subjects, we also determined the intrasubject variability of OFLX incorporation into individual hair strands. Four strands from each subject were segmented into 2-mm segments and analyzed. OFLX appeared in segments #1-#10 at week 5 (the first centimeter of hair). OFLX appeared in segments #2-#20 at week 7 (the first and second centimeter of hair). The maximum OFLX concentration (the "band" of drug) and location was then determined for each strand. The maximum OFLX concentration was measured in segments #2-#5 at week 5 for all subjects (within the first centimeter of hair length). The maximum OFLX concentration was measured in segments #3-#8 at week 7 (within the first and second centimeter of hair). This was consistent with a growth rate of less than 1.0 cm/month, although considerable intersubject variability was found. No significant axial diffusion of OFLX along the hair shaft beyond the first 3.0 cm of hair was noted. Despite a strong effect of hair color, these data suggest that OFLX may be a suitable marker substance for hair, allowing a subject to serve as their own "control". Future studies will explore whether drug use, treatment compliance, or recidivism in clinical drug-abuse studies can be determined with the aid of OFLX. PMID- 12731657 TI - 1H NMR spectroscopy and GC-MS analysis of alpha-chloralose. Application to two poisoning cases. AB - Alpha-chloralose, a compound widely used as a rodenticide and in the control of bird pests, is readily available. Two cases of intentional poisoning are reported. Both patients became comatose and presented hypersialorrhea and myoclonal crises in the legs. They were discharged from hospital after several days. As clinical signs of alpha-chloralose poisoning lack specificity, anamnesis might be difficult, particularly in the case of delayed diagnosis. Toxicological analysis is therefore critical, and this article reports the investigation of serum and urine samples by gas chromatography-mass spectrometry (GC-MS) in the electron-impact mode, and by 1H nuclear magnetic resonance (1H NMR) spectroscopy. Non-hydrolyzed urinary samples and those hydrolyzed by beta-glucuronidase were taken into consideration. After acetylation, GC-MS analysis was based on characteristic mass-to-charge ratio values of 272 for alpha-chloralose and 206 for beta-hydroxyethyltheophylline, which was used as internal standard. Characterization of alpha-chloralose species by 1H NMR spectroscopy was performed taking two parameters into account: chemical shift and coupling-constant values. Without any pretreatment, 1H NMR spectroscopy revealed the presence of free (5.50 and 6.15 ppm) and conjugated forms of alpha-chloralose by characteristic resonances of H1 and chloral-type protons, respectively. Quantitative analysis was performed by relative integration of peak areas. Serum alpha-chloralose showed concentrations below the quantitation limit of both methods. In urine samples, the free chemical species rapidly decreased. GC-MS analysis revealed the predominence of conjugation after a beta-glucuronidase hydrolysis step. 1H NMR analysis directly showed that on admission of the first patient, average urinary concentrations were 1.73 mmol/L (535 mg/L) for the free form and 13.72 and 6.25 mmol/L for the two conjugated forms. A later enzymatic treatment confirmed the total concentration of alpha-chloralose chemical species. Analysis of alpha chloralose in urine by either GC-MS or 1H NMR spectroscopy methods proved to be comparable. PMID- 12731658 TI - Determination of the herbicide glyphosate and its metabolite in biological specimens by gas chromatography-mass spectrometry. A case of poisoning by roundup herbicide. AB - In Japan, poisonings by the glyphosate (GLYP)-containing herbicide Roundup and the gluphosinate (GLUF)-based herbicide BASTA have been increasing since about 1987. We applied the gas chromatography-mass spectrometry (GC-MS) method of analysis, on which we have already reported in regard to the determination of the blood serum level of GLUF and its metabolite, for the determination of serum and urinary levels of GLYP and its metabolite aminomethyl phosphonic acid (AMPA). Derivatization using N-methyl-N-(tert-butyldimethylsilyl) trifluoroacetamide was completed at a temperature of 80 degrees C after 30 min, and the detection limit of GLYP was 10 pg using m/z 454 and that of AMPA was 1 pg using m/z 396. The full mass spectra of 100 pg GLYP and of 10 pg AMPA were obtained easily. In extractions for which the Isolute HAX cartridge was employed, the mean recovery rate of GLYP and AMPA added to serum to yield concentrations of 10-0.1 microg/mL (n = 5) was 91.6 +/- 10.6% (or better), whereas that of GLYP and AMPA added to urine to yield concentrations of 100-1.0 microg/mL (n = 10) was 93.3 +/- 6.6% (or better), both of which were good rates. Also, using this method of analysis, the presence of GLYP was identified in the full mass spectra obtained from the serum of a patient who may or may not have ingested Roundup. PMID- 12731659 TI - Time-adjusted urine/blood ratios of ethanol in drinking drivers. PMID- 12731660 TI - Comment on oral fluid testing for drugs of abuse: positive prevalence rates by Intercept immunoassay screening and GC-MS-MS confirmation and suggested cutoff concentrations. PMID- 12731661 TI - Comment on determination of glycols in biological specimens by gas chromatography mass spectrometry. PMID- 12731662 TI - Multistep tumorigenesis of multiple myeloma: its molecular delineation. AB - Multiple myeloma (MM) is an incurable malignant neoplasm affecting terminally differentiated B-cells. It derives from post-germinal center B-cells and develops as a result of multistep tumorigenic events, because approximately one third of all MM cases have a history of preceding monoclonal gammopathy of undetermined significance (MGUS) or smoldering myeloma. MM terminates in the formation of extramedullary invasion or in secondary plasma cell leukemia. To account for this clinical experience, investigators have found that intrinsic chromosomal instability followed by complex chromosomal translocations/deletions plays a crucial role in the development from MGUS to MM. Representative aberrations include chromosomal rearrangements involving 14q32 loci and deletion at the long arm of chromosome 13. Contributing to the progression of MM itself are genomic instability and altered methylation of the specific gene promoters. The former results in activation of specific oncogenes such as RAS and FGFR3 or in inactivation of p53, and the latter results in inactivation of tumor suppressor genes, including p16. An accurate understanding of each of these molecular events should help clarify the development of specific molecular targeting therapies based on the differences in dysfunctional signaling pathways found in the cells of all MM patients. PMID- 12731664 TI - Novel therapies in multiple myeloma. AB - The discovery of the activity of thalidomide in myeloma in the late 1990s transformed the therapy of myeloma dramatically. Apart from providing a useful treatment option for patients with myeloma, it has spurred clinical investigation of several other nonchemotherapeutic agents for this disease. These active, promising agents include CC-5013 (a thalidomide analog) and bortezomib (a proteasome inhibitor), as well as other agents, such as arsenic trioxide, ENMD 0995 and 2-methoxyestradiol. Preliminary data show that a number of these agents are active in treating disease that has relapsed after conventional chemotherapy as well as after high-dose therapy and transplantation, and some agents are active even after other novel agents have failed. The only novel drug that is commercially available currently is thalidomide, which has a therapeutically relevant benefit at all stages of the disease. A therapeutic trial of thalidomide is essential for all patients with myeloma. There are in vitro and in vivo data showing synergy between some of the novel agents. Although these novel drugs are typically used for treating disease that is refractory to or has relapsed after cytotoxic therapy, it is likely that they will start being used as part of frontline therapy, either by themselves or in combination with chemotherapy. PMID- 12731663 TI - Global gene expression profiling in the study of multiple myeloma. AB - Multiple myeloma (MM) is a rare but uniformly fatal malignancy of antibody secreting plasma cells. Although several key molecular events in disease initiation or progression have been confirmed (eg, FGFR3/MMSET activation) or implicated (eg, chromosome 13 deletion), the mechanisms of MM development remain enigmatic. Although it is generally indistinguishable morphologically, MM importantly exhibits a tremendous degree of variability in its clinical course, with some patients surviving only months and others for many years. However, measures of current laboratory parameters can account for no more than 20% of this outcome variability. Furthermore, the means by which current drugs impart their anti-MM effect are mostly unknown. The development of serious comorbidities, such as osteopenia and/or focal lytic lesions of bone, is also poorly understood. Finally, very little knowledge exists concerning the molecular triggers for the conversion of benign monoclonal gammopathy of undetermined significance (MGUS) to overt MM. Given that abnormal gene expression lies at the heart of most if not all cancers, high-throughput global gene expression profiling has become a powerful tool for investigating the molecular biology and clinical behaviors Here I discuss recent progress made in addressing many of these issues through the molecular dissection of the transcriptome of normal plasma cells, MGUS, and MM. PMID- 12731665 TI - Recent developments in hematopoietic stem cell transplantation for multiple myeloma. AB - Multiple myeloma (MM) is often successfully controlled with conventional chemotherapy; however, complete remissions are uncommon, and cure is rare. High dose therapy followed by administration of autologous or allogeneic stem cells, used for the treatment of MM in the past 15 years, is promising as a means of increasing remission rates and improving survival. Autologous transplantation has not always demonstrated survival benefits in randomized studies because most of the patients receiving transplants have relapses, whereas patients given conventional therapy can receive salvage transplants when relapse occurs. Efforts to improve the results of autologous transplantation include targeted radiation, tandem transplantation, and posttransplantation immunotherapy. Only allogeneic hematopoietic stem cell transplantation is potentially curative, owing to a graft versus-myeloma effect. Although patients who receive either allogeneic or autologous stem cell transplants for MM have similar 3- to 5-year survival rates, only allograft recipients appear to enjoy long-term disease-free survival. High transplantation-related mortality associated with allogeneic stem cell transplantation is currently the major limitation to wider use of this potentially curative modality. Strategies designed to improve the therapeutic index of allografts include the use of nonablative conditioning regimens, peripheral blood cells rather than bone marrow, graft engineering, and targeted conditioning therapies, such as bone-seeking radioisotopes. PMID- 12731666 TI - Can eradication therapy for Helicobacter pylori really improve the thrombocytopenia in idiopathic thrombocytopenic purpura? Our experience and a literature review. AB - Helicobacter pylori has recently been postulated to play a role in the pathogenesis of autoimmune diseases, including idiopathic thrombocytopenic purpura (ITP). We investigated the prevalence of H pylori infection and the effects of its eradication in 61 patients with ITP. H pylori infection was found in 50 patients (83%), an incidence significantly higher than not only healthy volunteers in Japan (60%) but also subjects in other reported ITP series (approximately 43%-71%). In our study, the mean age of H pylori-positive ITP patients (58.0 years) was significantly higher than that of H pylori-negative ITP patients (40.5 years). Bacterium eradication efforts were performed in 29 infected ITP patients and succeeded in 27 patients (93%). The 29 patients with eradicated H pylori infections showed significant increases in platelet counts compared with patients with uneradicated infections or who were H pylori negative. During the follow-up period (median, 11.0 months), 16 (55%) of 29 patients achieved a major or a minor response. The patients who achieved a major response had not received previous prednisolone therapy, suggesting a relationship between prednisolone therapy and the response to eradication efforts. The assessment of H pylori infection and its eradication should be attempted in cases of ITP, because this approach may be a good new strategy for treating some ITP patients, especially elderly Japanese patients. Some regional factors have been suggested as causes of H pylori-associated ITP. PMID- 12731667 TI - Tunica interna endothelial cell kinase expression and hematopoietic and angiogenic potentials in cord blood CD34+ cells. AB - Tunica interna endothelial cell kinase (TEK) is expressed in both hematopoietic and endothelial cells and plays a crucial role in hematopoiesis and angiogenesis in mouse development. In humans, however, little is known about the hematopoietic and angiogenic potentials of TEK-expressing cells in umbilical cord blood (CB) cells, which originate during the human fetal period. We therefore compared the hematopoietic and angiogenic abilities of CB CD34+TEK+ and CD34+TEK- cells by using a clonogenic assay and xenotransplantation into immunodeficient NOD/SCID mice. The results showed that colony-forming cells and cells capable of repopulating in NOD/SCID mice were present in both CD34+TEK+ and CD34+TEK- cells and that the hematopoietic activities of the cell types were similar. In contrast, the potential to differentiate into endothelial cells in vivo was greater in the CD34+TEK+ cells. All NOD/SCID mice engrafted with CD34+TEK+ cells had human CD31-expressing and VE-cadherin-expressing endothelial cells in the vessels of the ischemic muscles and/ or human endothelial cells expressing CD31, kinase-insert domain-containing receptor, and endothelial nitric oxide synthase in liver sinusoidal cells, whereas such endothelial cells were detected in only 3 of the 7 recipients engrafted with CD34+TEK- cells. This result has important implications in cell therapy using CB cells for treating hematopoietic disorders and vascular diseases. PMID- 12731668 TI - Serine protease inhibitors inhibit superoxide release and adherence in human neutrophils stimulated by granulocyte-macrophage colony-stimulating factor and tumor necrosis factor-alpha. AB - Stimulation of human neutrophils with granulocyte-macrophage colony-stimulating factor (GM-CSF) or tumor necrosis factor-alpha (TNF) results in increased superoxide (O2-) release and adherence. O2- release and adherence are dependent on activation of extracellular signal-regulated kinase (ERK) and p38 mitogen activated protein kinase (MAPK). Possible participation of serine proteases in GM CSF- or TNF-induced activation of human neutrophils was explored with various serine protease inhibitors, including phenylmethylsulfonyl fluoride, L-1 tosylamido-2-phenylethyl-chloromethyl ketone and N-alpha-p-tosyl-L-lysine chloromethyl ketone. GM-CSF- or TNF-induced O2- release and adherence were inhibited in parallel by pretreatment of neutrophils with these inhibitors. On the other hand, GM-CSF- or TNF-induced phosphorylation of ERK and p38 MAPK was unaffected by these inhibitors at the concentrations effective for the inhibition of O2- release and adherence. These findings suggest that serine proteases are involved in GM-CSF- and TNF-induced O2- release and adherence in human neutrophils and that serine proteases function downstream or independently of the activation of ERK and p38 MAPK. PMID- 12731669 TI - Absence of R24C mutation of the CDK4 gene in leukemias and solid tumors. AB - A mutation of the p16(INK4a)-binding domain of the cyclin dependent kinase 4 (CDK4) gene, R24C, has been reported in some cases of melanoma. This mutation prevented binding of the CDK4 inhibitor p16(INK48) to CDK4. To determine the relevance of the mutation, we performed polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analysis in diverse types of human leukemias and solid tumors. No mobility shifts indicating sequence alterations were observed in 273 tumors and 49 cell lines from diverse kinds of tumors These results suggest that in contrast to melanoma, in many other types of human neoplasms the mutation of the CDK4 gene is very rare. To better understand these findings, we randomly mutagenized the CDK4 gene and used the yeast two-hybrid method to screen for CDK4 mutants that had lost the ability to bind to p16(INK4a). Sequence analysis and in vitro kinase assays showed that most of the mutations that disrupted interactions with p16(INK4) also knocked out the activity of CDK4. This result may explain the rareness of CDK4 mutations in human tumors. PMID- 12731671 TI - Acute myelogenous leukemia M5b developed during clinical remission of Castleman disease. AB - Castleman disease (CD) is a rare heterogeneous lymphoproliferative disease characterized by clinical symptoms due to an excess of interleukin-6 (IL-6) or IL 6-like activity. We describe the first case of CD associated with acute myelogenous leukemia (AML). A 55-year-old man presented with skin rash on his face and multiple cervical lymphadenopathy. The results of examination of his lymph node biopsy specimen led to a diagnosis of CD. The symptoms resolved after the administration of prednisolone. Three years after the onset of CD, the patient's white blood cell count had increased to 63.4 x 10(9)/L. His bone marrow aspirate showed that approximately 80% of cells were leukemic, including well differentiated monocytic cells A diagnosis of AML M5b was made. The patient died of invasive pulmonary aspergillosis after chemotherapy. PMID- 12731670 TI - Constitutively activated Rho guanosine triphosphatases regulate the growth and morphology of hairy cell leukemia cells. AB - Hairy cell leukemia (HCL) is a rare type of chronic B-cell leukemia characterized by the hairy morphology of the leukemia cells. All of 5 HCL samples and an HCL derived cell line, BNBH-I, showed serrated edges and hairlike projections in May Grunwald Giemsa stain and protruding actin spikes and lamellipodia in phalloidin stain. These structures were hardly detected on B-cell chronic lymphocytic leukemia (B-CLL) and precursor B-cell acute lymphocytic leukemia (B-ALL) cells. Because Rho guanosine triphosphatases (GTPases) regulate the formation of these structures, we examined the expression levels and activation states of Rho GTPases in HCL cells. RhoA, Rac1, and Cdc42 were overexpressed and constitutively activated in HCL samples and BNBH-I cells but not in B-CLL or precursor B-ALL cells. Next we overexpressed dominant-negative (DN)-RhoA, DN-Rac1, and DN-Cdc42 in BNBH-I. As a result, each DN mutant repressed the growth of BNBH-I cells by more than 50% and inhibited actin spike formation, but only DN-Racl suppressed lamellipodia formation. We also found that enforced expression of constitutively active-RhoA, Rac, or Cdc42 in the proB-cell line Ba/F3 was sufficient to induce actin spike formation, whereas none of these molecules produced lamellipodia. These results indicated that constitutively activated Rho GTPases regulate the growth and unique morphology of HCL cells. PMID- 12731672 TI - Induction of complete remission of hypoplastic leukemia with antithymocyte globulin. AB - A 13-year-old boy was admitted to a local hospital because of pancytopenia. A bone marrow aspiration and biopsy revealed severely hypocellular marrow with no obvious leukemic cells. The diagnosis was severe aplastic anemia, and the patient was treated with antithymocyte globulin and cyclosporin A. A trilineage response was obtained, and the patient became transfusion-independent within 2 weeks. Two months later, the peripheral blood count normalized with an increased bone marrow cellularity. However, the patient was readmitted 5 months later for recurrence of the pancytopenia. A bone marrow aspiration revealed hypocellular marrow with morphologically blastoid cells. A surface marker study revealed the presence of a single clone that was positive for CD7, CD33, CD34, and HLA-DR. A diagnosis of hypoplastic leukemia was made on the basis of morphology and the surface marker studies. Retrospectively, the laboratory findings were the same as those seen at the onset of the disease. The patient did not respond to combination chemotherapy consisting of vincristine, prednisolone, cyclophosphamide, L-asparaginase, and doxorubicin, but administration of etoposide resulted in complete remission. An in vitro study revealed that >95% of the leukemic cells of this patient could be lysed after an incubation with antithymocyte globulin and human AB serum or baby rabbit serum. These findings suggest the efficacy of antithymocyte globulin in treating certain hypoplastic leukemias. PMID- 12731673 TI - Cytotoxic T-cell lymphoma arising in Behcet disease. AB - The case of a 49-year-old man with peripheral T-cell lymphoma arising in Behcet disease (BD) is reported. A diagnosis of incomplete BD was made, and the patient was treated with immunosuppressive agents for 9 months. A left perirenal mass emerged, and a computed tomography-guided needle biopsy of the tumor revealed the infiltration of small- and medium-sized lymphoma cells. The cells were positive for CD3, CD8, CD45RO, CD43, granzyme B, and T-cell intracellular antigen-1. A diagnosis of non-Hodgkin's lymphoma (diffuse medium, T-cell) was made. A left orbital mass also appeared. Standard combination chemotherapy diminished the perirenal and orbital lesions. Lymphoma cell infiltration in the esophagus was detected after chemotherapy, and the patient died of massive bleeding from the gastrointestinal tract. Non-Hodgkin's lymphoma is rarely associated with BD, and only 7 cases have been reported in the literature. We have summarized the published case reports of malignant lymphoma arising in BD. To our knowledge, this case report is the first to describe cytotoxic T-cell lymphoma arising in Behcet disease. PMID- 12731674 TI - The serum cytokine profiles of lymphoma-associated hemophagocytic syndrome: a comparative analysis of B-cell and T-cell/natural killer cell lymphomas. AB - To elucidate the differences in pathogenesis between lymphoma-associated hemophagocytic syndromes (LAHS) of the T-cell/ natural killer cell (T/NK) and B cell (B) types, we comparatively analyzed the clinical features and serum cytokine profiles of 33 patients with LAHS registered in the Kyoto University Hematology/Oncology Study Group. The serum cytokine levels of each patient group (B-LAHS versus T/NK-LAHS) were expressed as the ratio of the median to the upper normal values of the respective cytokines and were as follows: 19.05 versus 13.99 for soluble interleukin 2 (IL-2) receptor, 0.67 versus 0.67 for granulocyte macrophage colony-stimulating factor (GM-CSF), 0.64 versus 1.26 for G-CSF, 5.70 versus 3.61 for M-CSF, 1.54 versus 3.39 for interferon gamma (IFN-gamma), 13.17 versus 1.17 for IL-6, 6.88 versus 1.58 for tumor necrosis factor alpha (TNF alpha), 0.71 versus 0.41 for IL-1beta, 1.99 versus 0.21 for IL-12, and 105.32 versus 29.65 for IL-10. The serum levels of IL-6, TNF-alpha, and IL-10 were significantly higher in the B-LAHS group, whereas those of IFN-y were significantly lower. These differences between the 2 groups may reflect a difference in the pathogenesis Higher serum levels of IL-6, TNF-alpha, and IL-10 may be derived at least partly from neoplastic B-cells themselves In addition, the extremely high serum levels of IL-10 suggest that a compensatory anti inflammatory process may operate in both groups and give rise to a profound immunosuppressive state and a poor outcome. PMID- 12731675 TI - Hemophagocytic syndrome associated with severe adenoviral pneumonia: usefulness of real-time polymerase chain reaction for diagnosis. AB - In infection-associated hemophagocytic syndrome (HPS) the causative pathogen is often undetected, except in cases of herpes virus infections. We describe a 12 year-old girl with life-threatening pneumonia with HPS caused by an adenovirus. She was admitted with complaints of persistent fever and systemic petechiae/purpura. The day after admission the patient developed sudden dyspnea with massive infiltration of the bilateral lower lung field. She exhibited coagulopathy, hepatic dysfunction, and remarkable elevations in serum levels of cytokine, ferritin, and urinary beta2-microglobulin. A diagnosis of HPS was made, and the patient was treated with dexamethasone and cyclosporin A on the second hospital day. Her fever went down quickly, and the abnormal laboratory and chest radiographic findings returned to normal over a period of 2 weeks. Antibody analysis was not successful in identifying the pathogen responsible. However, a polymerase chain reaction (PCR) assay of lung tissue biopsied on the fifth hospital day was positive for an adenovirus (subgroup B), and quantitative adenoviral DNA analysis by real-time PCR using primers covering serotypes 3, 7,11, and 35 (all subgroup B) confirmed this initial finding (93 copies/microg DNA in the biopsied lung and no detectable adenovirus DNA in the lung tissues of control subjects). This approach may provide important clues for improving outcomes and clarifying the exact etiology in cases of often fatal, infection associated HPS. PMID- 12731676 TI - In vivo platelet activation and hyperaggregation in hemoglobin E/beta thalassemia: a consequence of splenectomy. AB - Patients with hemoglobin E/beta-thalassemia (E/beta-Thal) who have undergone splenectomy are prone to thrombosis in the small pulmonary arteries. To study the role of platelets in this situation, we assayed plasma beta2-thromboglobulin (betaTG) and performed whole blood platelet aggregation analysis of 30 E/beta Thal patients, half of whom had undergone splenectomy. We compared results with those obtained with 15 healthy control subjects. Plasma betaTG levels in splenectomy patients were significantly higher than in control subjects and patients who had not undergone splenectomy, and platelets in splenectomy patients exhibited hyperaggregation in response to adenosine diphosphate, thrombin, and ristocetin. Levels of plasma thrombin-antithrombin III complex were also significantly higher. This finding is likely due to an increased number of erythrocytes with exposed phosphatidylserines, an effect that has been associated with splenectomy. The increased presence of thrombin in the blood may well be the cause of platelet hyperactivity, which was evident only in the asplenic patients Platelet hyperactivity very likely plays a pathogenetic role in the thrombosis of small pulmonary arteries that occurs in E/beta-Thal patients who have undergone splenectomy. PMID- 12731677 TI - Serum immunoglobulin levels at onset: association with the prognosis of childhood idiopathic thrombocytopenic purpura. AB - Between 1981 and 2000, 87 patients with new diagnoses of idiopathic thrombocytopenic purpura (ITP) were admitted to the pediatric department of Kobe City General Hospital or Nishi-Kobe Medical Center. The patients' clinical records were analyzed for the relationships of disease outcome to serum immunoglobulin levels and other factors, including sex, onset age, and initial platelet counts. The disease of 22 patients became chronic, and of the 65 patients with an acute form of the disease, 27 exhibited levels of immunoglobulin G (IgG), IgA, or IgM above the 97.5% confidence limits of the age-matched control subjects. However, only 2 patients with the chronic form of the disease showed elevated serum immunoglobulin levels. The presence of antecedent specific viral infections was also associated with the acute disease form. In predicting the prognosis of childhood ITP, high serum immunoglobulin levels at initial presentation can be considered a good prognostic marker for the acute form of the disease. PMID- 12731678 TI - Pseudoprolongation of the prothrombin time during treatment with recombinant factor VIIa for bleeding in a patient with a spontaneous factor VIII inhibitor. PMID- 12731679 TI - Cognitive skills training: impact on drug abuse counseling and readiness for treatment. AB - As part of an NIDA-sponsored project, the current study investigated the effectiveness of a skills program we developed called the TCU Cognitive Skills Module (CSM). The program was developed to address cognitive deficits that may impede substance abuse treatment within the criminal justice system. Probationers in a 4-month residential program (followed by 3 months of aftercare) were randomly assigned (by community) to receive standard treatment or treatment enhanced with the CSM program. Peer ratings, individual ratings of the community, and counselor assessments revealed that the CSM was effective in increasing perceived treatment effectiveness (particularly midway through the program). There was also some support for the CSM being especially effective for probationers who enter treatment with lower levels of treatment readiness. PMID- 12731680 TI - Effectiveness of substance abuse treatment programming for women: a review. AB - Recent research has shown that women and men differ in substance abuse etiology, disease progression, and access to treatment for substance abuse. Substance abuse treatment specifically designed for women has been proposed as one way to meet women's distinctive needs and reduce barriers to their receiving and remaining in treatment. However, relatively few substance abuse treatment programs offer specialized services for women, and effectiveness has not been fully evaluated. This article reviews the literature on the extent and effectiveness of substance abuse treatment programming for women and provides an overview of what is known about the components of successful treatment programs for women. Thirty-eight studies of the effect on treatment outcomes of substance abuse treatment programming for women were reviewed. Seven were randomized, controlled trials, and 31 were nonrandomized studies. In our review, six components of substance abuse treatment programming for women were examined: child care, prenatal care, women-only programs, supplemental services and workshops that address women focused topics, mental health programming, and comprehensive programming. The studies found positive associations between these six components and treatment completion, length of stay, decreased use of substances, reduced mental health symptoms, improved birth outcomes, employment, self-reported health status, and HIV risk reduction. These findings suggest that to improve the future health and well-being of women and their children, there is a continued need for well designed studies of substance abuse treatment programming for women. PMID- 12731681 TI - Changes in health insurance and payment for substance use treatment. AB - Based on data from the National Household Survey on Drug Abuse in 1994 (N = 10, 158) and 1998 (N = 12,892) alcohol- or drug-using subsamples (aged 18-64) were compared to see if postwelfarereform reports of having employer-paid health insurance increased and Medicaid decreased by 1998 and affected participation in substance use treatment. By 1998, respondents were more likely to report having employer-paid insurance but reported source of payment for substance use treatment did not differ by year. In 1998, privately insured respondents were less likely to know if they had coverage for substance use treatment than in 1994. Of privately insured in treatment, 24% did not know if insurance covered treatment; most of these said they paid for treatment out-of-pocket. Despite reported increases in employer-paid insurance, uncertainty about its coverage of substance use treatment also increased and was associated with reports of not using it to pay for treatment. PMID- 12731682 TI - Self-reported health status among treated methamphetamine users. AB - Very little research has examined how drug abuse is related to general health status over the long term among both young and middle-aged adults. In this article, we investigate how self-reported health status is related to prolonged methamphetamine (MA) use in a diverse sample of MA users from ages 18 to 52 who have been treated for drug abuse in Los Angeles County. Using retrospective data, we investigate how prolonged MA use within younger and older age groups is related to two self-reported measures of current health status: the presence of a health condition that began after starting illegal drug use, and overall health. We control for the effects of drug use history, social and demographic factors, and other early experiences (e.g., early sexual abuse) that might be obstacles to achieving good health later in life. We find that having a current health condition is predicted by greater age and by more prolonged MA use, especially among younger people. Early sexual abuse predicts both measures of poor health. Current health status is predicted by several measures of drug use history and early experiences, but by fewer social and demographic factors. The results suggest that reduction of MA use among younger people is important in promoting their later health and that MA treatment services could be improved by a greater understanding of how early experiences influence later health. PMID- 12731683 TI - Community referral sources and entry of treatment-naive clients into outpatient addiction treatment. AB - This study assessed the association of sources of client referral with enrollment of treatment-naive clients. Data from the 1995 (n = 618) and 2000 (n = 745) waves of the National Drug Abuse Treatment Survey (DATSS), a panel study of outpatient substance abuse treatment units (OSAT), were analyzed. Enrollment of treatment naive clients was defined as the percentage of OSAT clients who entered treatment in the past 30 days with no prior treatment for substance abuse. A generalized estimating equation model simultaneously assessed the association of each referral source with the dependent variable, while controlling for potential confounding and accounting for correlation of unit-level responses over time. In the multivariable model, OSAT units with a greater proportion of treatment-naive clients had received more referrals from employee assistance programs and the criminal justice system, and fewer referrals from mental health agencies. No effect of referral from medical or social service agencies was observed. These results highlight the role of coercive community institutions in treatment outreach efforts to persons in earlier phases of the "addiction career." PMID- 12731684 TI - Factors influencing treatment enrollment by pregnant substance abusers. AB - Despite potentially devastating consequences to both mother and child, many pregnant substance abusers refuse treatment. To understand why, the present study compared women who enrolled in (N = 102) vs. declined (N = 23) day treatment. Participants were primarily African American, unemployed, high school graduates with a mean age of 27 years. Although demographic characteristics did not differ between groups, treatment enrollees had greater drug severity and were more likely to identify crack cocaine as their drug of choice. They also manifested more family and psychiatric problems, emotional distress, and Axis II psychopathology. In addition, treatment enrollees had greater legal severity and higher rates of criminal justice system involvement (46% vs. 13%). The women who declined perinatal addiction services may have perceived less need for intensive treatment due to lower problem severity and less distress. Development of alternative treatment approaches, such as limited perinatal addiction services provided in conjunction with prenatal care, is warranted. PMID- 12731685 TI - Case management for substance abusers with HIV/AIDS: a randomized clinical trial. AB - In a random assignment study, substance-abusing patients with HIV/AIDS in a public general hospital received a brief contact condition or received 12 months of case management delivered by paraprofessionals. Patient outcomes included substance use, HIV transmission risk, physical health, psychological status, and quality of living situation. In both conditions, a significant decrease occurred in a range of problems from Intake to the 6-month interview, followed by no significant pattern of change at 12- and 18-month interviews. On major outcome variables, there were no significant differences between the brief contact and case management conditions. Sixteen percent had died by the 18-month interview. Process data indicated wide variation in the amount of case management received by participants, and the amount of case management was not related to improvement in the outcome measures. The study has limitations yet does not support the hypothesis that case management improves outcomes better than brief contact for this population. PMID- 12731686 TI - Relationships between urinalysis testing for substance use, medical expenditures, and the occurrence of injuries at a large manufacturing firm. AB - Drug use among employees continues to be a serious concern for American employers. Over 80% of the large employers in the United States use some form of testing to detect drug use, but this practice is controversial and the cost effectiveness of drug testing remains largely unknown. This study begins an empirical investigation of the consequences of drug testing by estimating its impact on medical care expenditures and injury rates at a large manufacturing firm in 1996-1999. Multiple regression analyses of a pooled cross-sectional time series data set were used to separate the impact of drug testing from other factors and to help find the optimal level of testing that was associated with minimum medical expenditures. Results indicated that medical expenditures would be minimized when 42% of the employees in a calendar quarter were drug tested. This implies that, on average, employees should be tested 1.68 times a year. The results also indicated that doubling the testing rate would reduce the odds of incurring any injuries on the job by over half, but the injury rate was already so low that this impact was very small. Hopefully the results of this study will inform the policy debate over drug testing by focusing on real data, as opposed to supposition or political considerations that seem to dominate many discussions. PMID- 12731687 TI - Comparative profiles of women with PTSD and comorbid cocaine or alcohol dependence. AB - This study examined differences in substance abuse severity, trauma history, posttraumatic stress disorder (PTSD) symptomatology and psychiatric comorbidity among treatment-seeking women (N= 74) with PTSD and either comorbid cocaine or alcohol dependence. Women in the cocaine/PTSD group, compared with the alcohol/PTSD group, demonstrated greater occupational impairment (e.g., greater severity on the employment subscale of the Addiction Severity Index, less monthly income, fewer days worked in past month), more legal problems (e.g., greater number of months incarcerated and arrests for prostitution), and greater social impairment (e.g., fewer number of close friends, less likely to be married). Women in the alcohol/PTSD group evidenced higher rates of exposure to serious accidents, other situations involving serious injury, and other extraordinarily stressful life events. Rates of major depression and social phobia were higher among the alcohol/PTSD group than the cocaine/PTSD group. Women in the alcohol/PTSD group scored higher on the CAPS avoidance, hyperarousal, and total subscale scores. The current findings enhance our understanding of the substance specific profiles of women with PTSD and comorbid substance use disorders and may have important implications for the design of dual-diagnosis interventions. PMID- 12731688 TI - Level of response to alcohol measured on the self-rating of the effects of alcohol questionnaire in a group of 40-year-old women. AB - A low level of response (LR) to alcohol (i.e., the need for higher amounts to have an effect) is a genetically influenced characteristic that is both found in populations at high risk for future alcoholism and that predicts alcohol-related life problems in the future. A simple paper-and-pencil test that asks subjects to estimate the number of drinks required to produce each of four effects at different times in their lives, the Self-Rating of the Effects of Alcohol (SRE) questionnaire, was developed to facilitate studies of LR. However, data to date on this indirect measure of LR have focused primarily on young male populations. The current study reports SRE values from 121 middle class, middle-aged women who were selected because they were married to men who participated in a longitudinal study of sons of alcoholics and controls. As predicted, the SRE measure of LR, which records the perceived responses of alcohol early in the drinking career (FIRST 5), correlated significantly with recent drinks per drinking day (0.18, p < 0.05), a diagnosis of an alcohol use disorder (0.33, p < 0.001), and with a family history of alcohol dependence (0.20, p < 0.05). The LR value from the SRE reflecting the perceived number of drinks required for effects in the most recent 3 months correlated more closely with both quantity and frequency of drinking (0.36 and higher, p < 0.001), and related to recent nicotine use (0.23, p < 0.05). These data indicate the potential usefulness of the SRE as a measure of LR as a risk factor for alcoholism in relatively highly functioning middle-aged women. PMID- 12731689 TI - Predictors of drinking outcomes among alcoholics. AB - Predicting outcomes for individual patients entering substance abuse treatment has long been a clinical goal in the addictions field. Intake data from the Addiction Severity Index and other standardized scales were collected on 248 alcohol dependent/abusing patients entering an urban hospital treatment program. The outcome measure was frequency of drinking days in the past 30 days. Baseline data were used to identify predictors of posttreatment drinking frequency at two follow-up interviews (3 and 12 months postbaseline). Stepwise multiple regressions indicated that a set of baseline predictors accounted for similar and substantial proportions of outcome variance at the two follow-ups. When psychosocial predictors were combined with an index of alcohol use severity (which included drinking frequency), the proportions of variance explained were 31% and 28% at 3 and 12 months, respectively. Two psychosocial predictors were significant at both time periods, and thus most likely to be replicated in future research: a treatment motivation index (a combination of measures of commitment to treatment success and internal motivation to seek treatment) and an index of 12-step (self-help) participation (a combination of measures of frequency of 12 step meeting attendance and perceived helpfulness of 12-step participation). While the predictability of short-term (3 month) outcomes could help clinicians tailor treatment strategies to maximize patient motivation and reduce drinking behavior, the predictability of longer term (12 month) outcomes could help counselors plan aftercare programs, encourage self-help participation, and promote recovery-oriented activities to sustain initial treatment-induced gains. PMID- 12731690 TI - Blow and go: the Breath-Analyzed Ignition Interlock Device as a technological response to DWI. AB - Driving while intoxicated rates have declined substantially in the last 20 years. This is as a result of public opinion combined with increased law enforcement efforts. A recent tool has been the Breath Analyzed Ignition Interlock Device. This new technology is designed to prevent persons with excessive blood alcohol levels from operating the interlocked vehicle. This 3-year recidivism study of the ignition interlock revealed 17.5% recidivism rates for the interlock group compared to 25.3% recidivism rates for the non-interlock group, a 31% decrease. Multiple offenders and younger (under 30) offenders had significantly lower rates of subsequent arrests. The multi-offenders in the comparison group were more than twice as likely as the interlock group to have a subsequent conviction within 3 years. The difference was nearly the same for the under 30 age group. There was almost no difference for first offenders. Accordingly, the ignition interlock appears to significantly reduce recidivism for repeat and younger DWI offenders but offers almost no improvement for first offenders. One driver of 315 (0.32%) was charged with DWI with an interlock in place. This offender had a child provide the breath sample while she drove the vehicle. PMID- 12731691 TI - The efficacy of two prototype chewing gums for the removal of extrinsic tooth stain. AB - AIM: To compare the potential efficacy of two prototype chewing gums in extrinsic stain removal on natural teeth. SETTING: Dental school clinics. DESIGN: Double blind, two groups, parallel design. PARTICIPANTS: 76 adult volunteers (32m, 44f, mean age: 20.6 years old). METHODS: Oral hard and soft tissue health was examined. The subjects were randomly assigned to use either Product A (without active ingredients) or Product B (with active ingredients). Each subject was asked to brush their teeth for one minute twice daily (mornings and nights) and chew the gums supplied for 15 min (2 dragees each time), three times daily, once after each meal (breakfast, lunch and dinner) for the entire four-week duration. Dental stain assessment was made on the 12 incisors using the Lobene Stain Index (LSI). RESULTS: The overall difference between the stain scores after 4-weeks' use of the chewing gums was statistically significant (p<0.01) for both test Product A (10.84) and Product B (7.77) with regard to the mean baseline stain scores (21.57). This difference represented a 48% reduction in stain scores for those subjects using Product A, while the reduction was 64% for the subjects using Product B. CONCLUSIONS: The results of this in vivo study suggest that chewing gums with and without active ingredients have potential effect on stain removal after regular use for one month. PMID- 12731693 TI - Dental caries and periodontal status of 12-year-old school children in rural Qinghai, China. AB - OBJECTIVES: To describe the dental caries and periodontal status of 12-year-old children in rural Qinghai and to compare the oral health status between the Han and ethnic minority children. METHOD: Children aged 12-years in three primary schools in Hexi Townships, Guide County, Hainan Tibetan Autonomous State, Qinghai Province, China, were invited to participate in the survey conducted during July 2001. Each child was interviewed and clinically examined in the schools, using portable equipment. World Health Organization (WHO) examination procedures and diagnostic criteria were used. SPSS 11.0 was used for data compilation and statistical analysis. RESULTS: All 196 12-year-olds (118 Han and 78 from ethnic minorities, i.e. Muslims and Tibetans) were surveyed. 48% of the children reported never brushing their teeth, while 12% brushed only every 2-3 days. Over ninety per cent (n=182) of the children had never visited a dentist. The DMFT scores in Han and in minority groups were found to be 0.12 and 0.23 respectively. A larger proportion of children in Han group (94%) had no caries experience (DMFT = 0) than that of Muslim and Tibetan (82%) groups (p<0.01). None of the children had healthy gingiva (i.e. highest CPI score=0) and 85% had calculus. CONCLUSION: The prevalence of caries experience in Han children is significantly lower than that in Muslim and Tibetan children in rural Qinghai. Dental caries and caries treatment needs of Han and minority rural schoolchildren in Qinghai are low but their periodontal health status is unsatisfactory. Basic dental health care and preventive education for the children in this part of China is needed. PMID- 12731692 TI - Toothbrushing and gingival recession. AB - AIM: To review the hypothesis that toothbrushing leads to gingival recession. Gingival recession develops due to anatomical and pathological factors. The prevalence of recession is dependent on the age and characteristic of the population because it usually presents in individuals with periodontal disease or those who practise zealous or improper oral hygiene methods. Gingival trauma and gingival abrasion from toothbrushing is thought to progress directly to gingival recession. Case studies documenting recession from toothbrush trauma are speculative. Short-term studies suggest that gingival trauma and gingival abrasion may result from toothbrushing, but the direct relationship between traumatic home care and gingival recession is inconclusive. Long-term studies remain elusive or do not support the development of recession following toothbrushing. Nevertheless, tooth abrasion may be an integral part in the aetiology of recession. Toothbrush abrasion also may cause wear at the cemento enamel junction resulting in the destruction of the supporting periodontium leading to recession. PMID- 12731694 TI - An assessment of relationship between self-esteem, orthodontic concern, and Dental Aesthetic Index (DAI) scores among secondary school students in Ibadan, Nigeria. AB - OBJECTIVE: To ascertain the correlation between self-esteem, orthodontic concern and orthodontic status using DAI scores in a group of Nigerian potential orthodontic patients. DESIGN: A cross-sectional analytical study. SETTING: Secondary schools reflecting a good socio-economic spread of adolescents in the town. PARTICIPANTS: Randomly selected 520 junior and senior students 276 (53.1%) males and 244 (46.9%) females with mean age of 15.02 +/- 3.26 (SD) years participated in the study. METHODS: Each student was asked to fill in the questionnaire on orthodontic concern and Global Negative Self-Evaluation scale (GSE) with subsequent assessment of the occlusion according to the DAI by one orthodontist. After obtaining consent, thirty students were re-examined to test intra-examiner reliability which was good (r = 0.98, P < 0.001). RESULTS: The correlations between self-esteem, orthodontic concern and DAI scores were tested using Spearman rank order correlation coefficient. Significant positive correlations were observed between self-esteem and orthodontic concern according to DAI treatment category needs (r = 0.274, p<0.01; r = 0.396, p<0.01; r = 0.347, p<0.05) except for the severe malocclusion group which was positive but not statistically significant (r = 0.136, p> 0.05). Significant correlation was equally noted between DAI scores and orthodontic concern (r = 0.191, p<0.01). CONCLUSION: Significant positive correlation existed between self-esteem in a group of Nigerian adolescents and their orthodontic concern as well as between their DAI scores and orthodontic concern. DAI should be a relevant and useful occlusal index for the Nigerian orthodontic population. PMID- 12731695 TI - Neuropsychological effects of low mercury exposure in dental staff in Erzurum, Turkey. AB - OBJECTIVE: To carry out measurements of the Hg levels and personal exposure in Turkish dental clinics, and to evaluate possible adverse effects on the CNS in dental personnel. SETTING: Five dental clinics (1 private, 4 public) in Erzurum, Turkey. SUBJECTS AND METHODS: 43, Hg vapour-exposed dental staff were examined and 43 hospital employees with no known exposure to Hg acted as the control group. Hg concentrations in plasma and urine were analysed by atomic absorption spectrophotometry. Possible effects on the central nervous system (CNS) were estimated by neuropsychological tests (Weschler Memory Scale-Revised (WMS-R) and Verbal Test of Memory Processes (VTMP)) and two self-administered questionnaires (Symptom Checklist-90-Revised (SCL-90-R) and Beck Depression Inventory (BDI)). RESULTS: The dental staff group had higher whole blood (B-Hg) and urine (U-Hg) Hg levels than the control group. The mean B-Hg value was 2.18 nmol/l and U-Hg was 1.17 nmol/mmol creatinine. U-Hg had an inverse relationship with logical memory (in WMS-R test) and total retention score (in VTMP test), and a positive relationship with increased scores of Anxiety and Psychoticism (in SCL-90-R). CONCLUSION: These results may represent long-term consequences of low Hg exposure. In dentistry, to decrease toxic effects, proper Hg hygiene should be practiced by all dental health care workers. PMID- 12731696 TI - Periodontal health among recent immigrants from Quara, rural Ethiopia and indications for periodontal health care. AB - OBJECTIVES: To investigate periodontal status and oral hygiene practices among recent new immigrants from Quara, Ethiopia, with the aim to maintain and promote their periodontal health. METHODS: Periodontal status was recorded for a total of 487 participants using the Community Periodontal Index (CPI). Age was grouped into 13-17, 18-24, 25-34, 35-44, and 45-65 years, and its association with 'worst CPI' was tested. Information on oral hygiene behaviour was collected by interview. RESULTS: The percentage of subjects with calculus is most common in all age groups. Low levels of periodontal pockets in the presence of calculus and very low levels of deep pockets were detected. Differences in CPI scores among the various age groups were statistically significant (p<0.0001). Oral hygiene practice is rare among children in Ethiopia, but a considerable proportion (53%) immediately adopted the local common custom and started brushing their teeth in Israel. Among the adults, oral hygiene practice with chewing sticks was common. CONCLUSIONS: Periodontal health promotion of this population should be targeted towards the entire population by enhancement of self-care oral hygiene measures. Periodontal treatment of this population, especially scaling and root planing remains controversial. The presence of inferior oral hygiene and high levels of calculus, together with low levels of periodontal pathology, raises important thoughts regarding the natural history of periodontal disease development. PMID- 12731697 TI - Restorative care and economic wealth: a global perspective. AB - There are increasing numbers of reports of inequalities in access to, and the provision of, restorative care that may be related to economic factors. This brief communication reports an investigation as to whether a relationship may be established between the per capita gross domestic product (GDP), the average number of filled teeth (FT) and the Care Index (CI) for 35-44-year-old adults in different countries. It is concluded that a direct proportional relationship does exist for the countries investigated in this study. PMID- 12731698 TI - Guidance of eruption for general practitioners. AB - The principle of early treatment through well-planned extraction of primary teeth followed by removal of permanent teeth has stood the test of time. The objective of this article is to develop some simple guidelines for general dental practitioners to perform 'guidance of eruption' in malocclusion with severe crowding. PMID- 12731699 TI - Severe acute respiratory syndrome (SARS) and coronavirus testing--United States, 2003. AB - CDC and the World Health Organization (WHO) are continuing to investigate the multicountry outbreak of severe acute respiratory syndrome (SARS). Infection with a novel coronavirus has been implicated as a possible cause of SARS. This report updates information on U.S. residents with SARS and summarizes the clinical histories of the five U.S. residents identified as of April 9, 2003, who have both suspected SARS and laboratory evidence of infection with a novel coronavirus. PMID- 12731700 TI - Prevalence of current cigarette smoking among adults and changes in prevalence of current and some day smoking--United States, 1996-2001. AB - Tobacco use, particularly cigarette smoking, is the leading preventable cause of death in the United States and is responsible for approximately 440,000 deaths each year. One of the national health objectives for 2010 is to reduce the prevalence of cigarette smoking among adults to < or = 12% (objective 27.1). To examine the prevalence of smoking for the 50 states, the District of Columbia (DC), Guam, Puerto Rico, and the Virgin Islands, CDC analyzed data from the 2001 Behavioral Risk Factor Surveillance System (BRFSS). This report summarizes the results of that analysis, which indicate that, during 2001, the median adult current smoking prevalence was 23.4% (range: 13.3%-30.9%) for the states and DC, and 12.5% (range: 9.8%-31.4%) for Guam, Puerto Rico, and the Virgin Islands. During 1996-2001, the prevalence of current smoking was relatively stable in 41 states and DC, and the proportion of current smokers who were some day smokers increased significantly in 31 of those states and DC. Because the only safe alternative to smoking is cessation, interventions should target all smokers to help them quit smoking completely. PMID- 12731701 TI - Observational survey of smoking provisions in food service establishments- Southeast Health District, Georgia, 2001. AB - In the United States, approximately 38,000 deaths are attributable to second hand smoke (SHS) exposure each year. One of the national health objectives for 2010 is to reduce public exposure to environmental tobacco smoke (ETS) (objective 27.10). To reduce public exposure to ETS, CDC recommends smoking bans and restrictions in public places. Some of the highest reported exposures to concentrations of SHS are found in food service establishments; however, Georgia does not have a state law prohibiting smoking in these places. In March 2001, the director of Georgia's Southeast Health District requested assistance from the state health department in developing a surveillance system of smoking provisions in food service establishments. This report summarizes an observational survey of smoking provisions in food service establishments of Georgia's Southeast Health District in 2001, which found that although 69.4% (506) of all surveyed establishments were completely smoke free, the remaining establishments failed to provide several physical modifications designed to minimize ETS exposure. Public health officials in the Southeast Health District will use survey results to target interventions toward establishments lacking ETS-minimizing provisions. PMID- 12731702 TI - Point-of-purchase alcohol marketing and promotion by store type--United States, 2000-2001. AB - Alcohol consumption is the third leading preventable cause of death in the United States, accounting for approximately 100,000 deaths annually. Efforts to reduce the adverse health and social consequences from alcohol use include policies to restrict access to alcohol among underaged persons (i.e., persons aged <21 years) and to reduce alcohol-impaired driving among persons of all ages. Recent studies have focused on alcohol marketing as a potentially important contributor to alcohol consumption, particularly among underage drinkers. Point-of-purchase (POP) (i.e., on-site) marketing, including alcohol advertising and placement, can increase alcohol sales and consumption substantially, thereby increasing the risk for various alcohol-related health outcomes, including alcohol-impaired driving and interpersonal violence. To assess the type and frequency of POP alcohol marketing, researchers with the ImpacTeen Project collected and analyzed store observation data during 2000-2001 from 3,961 alcohol retailers in 329 communities throughout the United States. This report summarizes the results of the study, which indicate that POP alcohol marketing is extensive in certain store types frequented by teenagers and young adults. Public health agencies and policy makers should work with liquor control boards to reduce POP marketing that could promote risky or underage drinking. PMID- 12731703 TI - Update on adverse events following civilian smallpox vaccination--United States, 2003. AB - During January 24-April 4, 2003, smallpox vaccine was administered to 31,297 civilian health-care and public health workers in 54 jurisdictions as part of an effort to prepare the United States for a possible terrorist attack using smallpox virus. This report updates information on all vaccine-associated adverse events among civilians vaccinated since the beginning of the smallpox vaccination program and among contacts of vaccinees, received by CDC from the Vaccine Adverse Event Reporting System (VAERS) as of April 4. PMID- 12731704 TI - Individual differences in children's event memory reports and the narrative elaboration technique. AB - Children between 7 and 8 years old took part in a staged event at school and 1 week later were assessed using a short form of the Wechsler Intelligence scale for children (third edition) and measures of metamemory, narrative ability, and socioeconomic status. Two weeks following the event, children either received narrative elaboration training (NET; K.J. Saywitz & L. Snyder, 1996) and were prompted with the four NET cue cards at interview; received verbal prompts corresponding to the cue card categories, but without prior training; or were presented with the cards at interview without prior training. Children given verbal labels as prompts recalled as much information as children who received NET training and cue cards. Measures of intelligence were predictive of amount recalled for cards-only children but not for the other 2 groups, indicating that differences in recall between low- and high-IQ groups were attenuated when recall was supported by NET training or verbal prompting. PMID- 12731705 TI - Predicting unit performance by assessing transformational and transactional leadership. AB - How do leadership ratings collected from units operating under stable conditions predict subsequent performance of those units operating under high stress and uncertainty? To examine this question, the authors calculated the predictive relationships for the transformational and transactional leadership of 72 light infantry rifle platoon leaders for ratings of unit potency, cohesion, and performance for U.S. Army platoons participating in combat simulation exercises. Both transformational and transactional contingent reward leadership ratings of platoon leaders and sergeants positively predicted unit performance. The relationship of platoon leadership to performance was partially mediated through the unit's level of potency and cohesion. Implications, limitations, and future directions for leadership research are discussed. PMID- 12731706 TI - Culture moderates the self-regulation of shame and its effects on performance: the case of salespersons in The Netherlands and the Philippines. AB - In this study, the authors investigated how salespeople within an interdependent based culture (the Philippines) and an independent-based culture (the Netherlands) experience and self-regulate shame. Filipino and Dutch employees were found to experience shame as a consequence of customer actions in largely similar ways (i.e., for both, shame is a painful self-conscious emotion with unique physiological/behavioral urges, self-focused attention, and felt threat to the core self) but have different responses to their felt shame. Specifically, shame is self-regulated dissimilarly in the 2 cultures and leads to opposite effects on performance, namely, enhanced customer relationship building and civic virtue and helping occur for Filipino employees, and diminished sales volume, communication effectiveness, and relationship building transpire for Dutch employees. The positive effects experienced by Filipino employees occur through direct responses to felt shame and as a result of adaptive resource utilization. The negative effects experienced by Dutch employees occur as a result of the dysfunctional (from the firm's point of view) discharge of protective actions. PMID- 12731707 TI - Effectiveness of training in organizations: a meta-analysis of design and evaluation features. AB - The authors used meta-analytic procedures to examine the relationship between specified training design and evaluation features and the effectiveness of training in organizations. Results of the meta-analysis revealed training effectiveness sample-weighted mean ds of 0.60 (k = 15, N = 936) for reaction criteria, 0.63 (k = 234, N = 15,014) for learning criteria, 0.62 (k = 122, N = 15,627) for behavioral criteria, and 0.62 (k = 26, N = 1,748) for results criteria. These results suggest a medium to large effect size for organizational training. In addition, the training method used, the skill or task characteristic trained, and the choice of evaluation criteria were related to the effectiveness of training programs. Limitations of the study along with suggestions for future research are discussed. PMID- 12731708 TI - The two faces of transformational leadership: empowerment and dependency. AB - Followers' identification with the leader and the organizational unit, dependence on the leader, and empowerment by the leader are often attributed to transformational leadership in organizations. However, these hypothesized outcomes have received very little attention in empirical studies. Using a sample of 888 bank employees working under 76 branch manages, the authors tested the relationships between transformational leadership and these outcomes. They found that transformational leadership was positively related to both followers' dependence and their empowerment and that personal identification mediated the relationship between transformational leadership and followers' dependence on the leader, whereas social identification mediated the relationship between transformational leadership and followers' empowerment. The authors discuss the implications of these findings to both theory and practice. PMID- 12731709 TI - Personality and the goal-striving process: the influence of achievement goal patterns, goal level, and mental focus on performance and enjoyment. AB - The purpose of this study is to examine the mechanisms by which personality traits influence performance and satisfaction. Specifically, the authors examined how 3 personality characteristics derived from self-determination theory (autonomy, control, and amotivated orientations) influence performance and enjoyment through achievement goal patterns, goal level, and mental focus. Data were collected from 284 students at 5 points in time. In particular, mental focus emerged as an important aspect of the self-regulation process. The results suggest that global personality traits can help researchers to understand and predict the motivational strategies that people use while working toward goals in achievement settings. PMID- 12731710 TI - A longitudinal analysis of organizational fairness: an examination of reactions to tenure and promotion decisions. AB - Most organizational justice research takes a cross-sectional approach to examining the relationship between perceived fairness and individuals' attitudes. This study examines the effect of procedural and distributive justice over time. It is suggested that individuals acquire more information and experience with procedures and outcomes over time. These changes in information and experience affect the influence of procedural and distributive justice on organizational attitudes. Faculty perceptions of tenure and promotion decisions were assessed 3 times (preallocation, short-term postallocation, long-term postallocation) over a 2-year period. Results generally supported the hypotheses. Procedural justice was most influential prior to and soon after outcome decisions were made. Distributive justice was most influential 1 year later. PMID- 12731711 TI - High-quality work, job satisfaction, and occupational injuries. AB - The authors investigated whether and how 1 element of a high-performance work system, namely high-quality jobs (composed of extensive training, variety, and autonomy), affects occupational injuries. On the basis of data from the Australian WIRS95 database (N = 16,466; Department of Workplace Relations and Small Business, 1997), high-quality jobs exerted a direct effect on injuries and an indirect effect through the mediating influence of job satisfaction. Conceptual, methodological, and practical issues are discussed. PMID- 12731712 TI - Antecedents and consequences of emotional display rule perceptions. AB - Central to all theories of emotional labor is the idea that individuals follow emotional display rules that specify the appropriate expression of emotions on the job. This investigation examined antecedents and consequences of emotional display rule perceptions. Full-time working adults (N = 152) from a variety of occupations provided self-report data, and supervisors and coworkers completed measures pertaining to the focal employees. Results using structural equation modeling revealed that job-based interpersonal requirements, supervisor display rule perceptions, and employee extraversion and neuroticism were predictive of employee display rule perceptions. Employee display rule perceptions, in turn, were related to self-reported job satisfaction and coworker ratings of employees' emotional displays on the job. Finally, neuroticism had direct negative relationships with job satisfaction and coworker ratings of employees' emotional displays. PMID- 12731713 TI - Organization structure as a moderator of the relationship between procedural justice, interactional justice, perceived organizational support, and supervisory trust. AB - Organizational justice researchers recognize the important role organization context plays in justice perceptions, yet few studies systematically examine contextual variables. This article examines how 1 aspect of context- organizational structure--affects the relationship between justice perceptions and 2 types of social exchange relationships, organizational and supervisory. The authors suggest that under different structural conditions, procedural and interactional justice will play differentially important roles in determining the quality of organizational social exchange (as evidenced by perceived organizational support [POS]) and supervisory social exchange (as evidenced by supervisory trust). In particular, the authors hypothesized that the relationship between procedural justice and POS would be stronger in mechanistic organizations and that the relationship between interactional justice and supervisory trust would be stronger in organic organizations. The authors' results support these hypotheses. PMID- 12731714 TI - The impact of psychological distress on absence from work. AB - The aims of this study were to examine the impact of psychological distress on absence from work; to compare psychological distress and job satisfaction as predictors of absence; and to compare the relative effects of 2 components of psychological distress, depression and anxiety, on absence. Organizational records of absence over a 3-year period were obtained for 323 health service staff in the United Kingdom, who also completed self-report measures of psychological distress and job satisfaction at the beginning of the study and 2 years later. Psychological distress, particularly depression, was found to predict absence, with higher levels of distress predicting a greater number of days and number of times absent. Job satisfaction and psychological distress independently predicted levels of absence. The psychological distress-absence relationship was not moderated by demographic variables. PMID- 12731715 TI - Descriptions and identifications of strangers by youth and adult eyewitnesses. AB - Two studies varying target gender and mode of target exposure were conducted to compare the quantity, nature, and accuracy of free recall person descriptions provided by youths and adults. In addition, the relation among age, identification accuracy, and number of descriptors reported was considered. Youths (10-14 years) reported fewer descriptors than adults. Exterior facial descriptors (e.g., hair items) were predominant and accurately reported by youths and adults. Accuracy was consistently problematic for youths when reporting body descriptors (e.g., height, weight) and interior facial features. Youths reported a similar number of descriptors when making accurate versus inaccurate identification decisions. This pattern also was consistent for adults. With target-absent lineups, the difference in the number of descriptors reported between adults and youths was greater when making a false positive versus correct rejection. PMID- 12731716 TI - Receiving instrumental support at work: when help is not welcome. AB - Although the role of social support in promoting employees' health and well-being has been studied extensively, the evidence is inconsistent, sometimes even suggesting that social support might have negative effects. The authors examined some psychological processes that might explain such effects. On the basis of the threat-to-self-esteem model, the authors tested the hypothesis that receiving imposed support elicits negative reactions, which are moderated by someone's need for support. The authors distinguished 3 different reactions: (a) self-related, (b) interaction-related, and (c) physiological. The results of an experiment with 48 temporary administrative workers generally confirmed the hypothesis. Imposed support elicited negative reactions, except when there was an unsolvable problem, but even then the effect of imposed support was not positive but neutral. PMID- 12731717 TI - The team halo effect: why teams are not blamed for their failures. AB - In this study, the existence of the team halo effect, the phenomenon that teams tend not to be blamed for their failures, is documented. With 2 studies using both real teams and controlled scenarios, the authors found evidence that the nature of the causal attribution processes used to diagnose failure scenarios leads to individuals being more likely to be identified as the cause of team failure than the team as a collective. Team schema development, as indexed by team experience, influences this effect, with individuals who have more team experience being less likely to show the team halo effect PMID- 12731718 TI - Improving the identification accuracy of senior witnesses: do prelineup questions and sequential testing help? AB - Eyewitness research has identified sequential lineup testing as a way of reducing false lineup choices while maintaining accurate identifications. The authors examined the usefulness of this procedure for reducing false choices in older adults. Young and senior witnesses viewed a crime video and were later presented with target present orabsent lineups in a simultaneous or sequential format. In addition, some participants received prelineup questions about their memory for a perpetrator's face and about their confidence in their ability to identify the culprit or to correctly reject the lineup. The sequential lineup reduced false choosing rates among young and older adults in target-absent conditions. In target-present conditions, sequential testing significantly reduced the correct identification rate in both age groups. PMID- 12731719 TI - Faking and selection: considering the use of personality from select-in and select-out perspectives. AB - The effects of faking on criterion-related validity and the quality of selection decisions are examined in the present study by combining the control of an experiment with the realism of an applicant setting. Participants completed an achievement motivation measure in either a control group or an incentive group and then completed a performance task. With respect to validity, greater prediction error was found in the incentive condition among those with scores at the high end of the predictor distribution. When selection ratios were small, those in the incentive condition were more likely to be selected and had lower mean performance than those in the control group. Implications for using personality assessments from select-in and select-out strategies are discussed. PMID- 12731720 TI - Accurate tests of statistical significance for r(WG) and average deviation interrater agreement indexes. AB - The authors demonstrated that the most common statistical significance test used with r(WG)-type interrater agreement indexes in applied psychology, based on the chi-square distribution, is flawed and inaccurate. The chi-square test is shown to be extremely conservative even for modest, standard significance levels (e.g., .05). The authors present an alternative statistical significance test, based on Monte Carlo procedures, that produces the equivalent of an approximate randomization test for the null hypothesis that the actual distribution of responding is rectangular and demonstrate its superiority to the chi-square test. Finally, the authors provide tables of critical values and offer downloadable software to implement the approximate randomization test for r(WG)-type and for average deviation (AD)-type interrater agreement indexes. The implications of these results for studying a broad range of interrater agreement problems in applied psychology are discussed. PMID- 12731721 TI - Remembering Arthur Kornhauser: industrial psychology's advocate for worker well being. AB - Arthur Kornhauser was an early industrial psychologist whose contributions have been neglected in written histories of applied psychology. Throughout his career, he was a staunch advocate for an industrial psychology that concentrated on improving workers' lives. This article describes his contributions to improving worker well-being in the research areas of testing, employee attitude surveying, labor unions, and mental health of workers. His most enduring quality was his outspoken advocacy for an industrial psychology that addressed workers' issues instead of management's prerogatives. On the basis of Kornhauser's accomplishments, a case can be made for him as one of industrial and organizational psychology's most important early figures. PMID- 12731722 TI - A patient with metachronous gastric, colonic, and thyroid cancers: a case report. AB - In this paper, we discuss a 62-year-old woman who, in the course of 7 years (1994 2001), developed three cancers: adenocarcinoma of the stomach, adenocarcinoma of the cecum, and insular thyroid carcinoma, which metastasized to the retroperitoneal lymph nodes and liver. The patient died from complications related to the metastases. The results of basic genetic tests were normal. To the best of our knowledge, no other patient with the combination of these three cancers has been reported in the literature. Although patients with multiple cancers are not common, it is nonetheless important for clinicians to consider the possibility of second and third cancers in patients who were treated for a primary malignant tumor. PMID- 12731723 TI - Leiomyosarcoma of the ovarian vein: report of a case. AB - Venous leiomyosarcomas are rare tumors, generally originating from the inferior vena cava. Only two cases of ovarian vein leiomyosarcoma have been reported thus far. Herein we report the case of a 50-year-old woman with a leiomyosarcoma of the right ovarian vein. The patient underwent radical en bloc extirpation of the tumor followed by radiation therapy. The clinicopathologic features and treatment are discussed. PMID- 12731724 TI - Postoperative colonic perforation in a right-sided congenital diaphragmatic hernia in an adult. AB - Congenital posterolateral hernias rarely present in the adult. We describe an unusual case that presented after a routine total abdominal hysterectomy. The patient developed acute respiratory symptoms caused by a perforated colon and hypoplastic right hemi-liver contained within a right-sided hernia. There was also an associated hypoplastic right lung. The hernia was repaired through a midline laparotomy, with no long-term complications. PMID- 12731725 TI - Post-traumatic hepatic artery aneurysm--surgical treatment: report of a case. AB - Hepatic artery aneurysms (HAAs) are uncommon. We present the diagnosis and surgical treatment of a post-traumatic, extrahepatic artery aneurysm. PMID- 12731726 TI - Extrahepatic division of the right hepatic vein in right hepatectomy for blunt liver trauma. AB - During a 5-year period, 10 patients with right-side blunt liver injury received an anatomic liver resection, using the technique of extrahepatic right hepatic vein division before hepatectomy. Five patients required a right posterior sectionectomy and 5 patients needed a right hemihepatectomy. The mean operation time was 146 minutes and median perioperative blood transfusion was 21.5 units (range, 4-94 units, packed red blood cell (PRBC) or whole blood). There was one hospital mortality; the patient died from multiple organs failure on postoperative day 5. Postoperative complications occurred in 4 of 9 surviving patients. Liver-related morbidity occurred in 2 patients who required reoperation for evacuation of subphrenic hematoma. The median hospital stay was 17 days (range, 6-84 days). Application of extrahepatic division of the right hepatic vein during a right hepatectomy for blunt liver trauma has several advantages: compression of a traumatic liver during mobilization; shortening of the ischemia duration; complete mobilization of the liver before parenchymal resection; and prevention of oozing with easier approximation of the raw surface. PMID- 12731727 TI - Anorectal melanoma: a large clinicopathologic study from India. AB - Anorectal melanoma is a rare tumor. It has dismal prognosis; only 10% of patients live longer than 5 years. It commonly presents as bleeding from the rectum or as a hemorrhoidal mass. The management of this tumor is controversial. Some authors believe that a palliative local excision is the treatment of choice, whereas others recommend radical surgery in localized disease. We present a retrospective analysis of 72 patients who were managed at our center between 1990 and 2001. This is the largest series from India. Most patients (48/72; 66%) presented with distant metastases. Twenty-four patients (24/72; 33%) underwent an abdomino perineal resection. Nineteen patients (19/24) had positive lymph node disease, and the mean disease-free survival in these patients was 10.3 months. Disease free survival in the node-negative patients was 26.5 months. A subset of patients with localized disease can benefit from radical surgery. In patients with large bulky localized disease, radical surgery provides better palliation than local excision. PMID- 12731728 TI - Ets-1 expression in vascular endothelial cells as an angiogenic and prognostic factor in colorectal carcinoma. AB - We investigated Ets-1 expression and its influence on angiogenesis and survival using immunohistochemistry of surgical specimens from 95 patients of colorectal cancer. The expression rate of Ets-1 was 48.4% of all cases in cancer cells, 65.3% in endothelial cells, and 28.4% in stromal cells. The microvessel density in patients with vascular Ets-1 expression was significantly higher than that in patients without the expression. A significant positive correlation was discovered between the Ets-1 vascular expression and vascular endothelial growth factor (VEGF) expression, or pyrimidine nucleoside phosphorylase (dThdPase) activity in the tumor. The positivity of vascular Ets-1 expression was higher in node-positive patients and in M1 patients. The survival rate was significantly lower in those cases positive for vascular Ets-1. Angiogenic factors such as VEGF might be involved in tumor angiogenesis through the Ets-1 vascular expression. The Ets-1 vascular expression may be a prognostic factor for colorectal cancer. PMID- 12731730 TI - Carcinoid tumor in a Meckel's diverticulum: hypothesis on mutual embryological origin. AB - Congenital defects presenting at an adult age are subject to diagnostic errors because of their relative rarity and often odd clinical presentation. We illustrate a 63-year-old male patient with multiple ileal carcinoid tumors along with a carcinoid tumor in Meckel's diverticulum. The Meckel's diverticulum is a congenital abnormality arising from a patent vitelline duct and is found at the anti-mesenteric side of the ileum. In the Meckel's diverticulum, ectopic tissue or neoplasms are sometimes found, which may lead to intussusception, hemorrhage, or inflammation. The carcinoid tumor is a neuroendocrine neoplasm originating from the neural crest and is diagnosed at virtually all ages. It may exhibit malignant behavior but generally has a mild clinical course. It is most often found in endodermally derived organs, especially the digestive and pulmonary tracts, and coexists frequently with malignant tumors, predominantly of endodermal origin such as colorectal carcinoma. The carcinoid tumor and Meckel's diverticulum are known to coincide, whereas the carcinoid is not really considered to be an embryologically determined defect. We hypothesize that both lesions are derived from local erroneous interaction among the neural crest and the endoderm. PMID- 12731729 TI - Giant diverticulum of the colon: report of two new cases and review of the literature. AB - Diverticulosis coli affects more than one in three individuals older than 65 in the Western world. Giant diverticulum of the colon is an extremely rare complication of diverticular disease; only 113 cases, mostly situated in the colon sigmoideum, have been reported in the world literature. Two new cases of giant diverticulum of the colon sigmoideum, with totally different clinical presentation, diagnosis, and management, are reported-one being the cause of chronic anemia and the other presenting as an acute abdomen. Based on a review of the literature, an update on symptomatology, diagnosis, pathogenesis, and therapeutical options of this rare disorder is provided. PMID- 12731731 TI - Effect of FK506 and basic fibroblast growth factor on nerve regeneration using a polytetrafluoroethylene chamber for nerve repair. AB - Peripheral nerve repair can be accomplished by using a polytetrafluoroethylene tubular chamber to guide nerve healing and regeneration. In this study, we delivered basic fibroblast growth factor (bFGF) into the chamber for sciatic nerve repair in rats. In addition, the animals were given systemically 1 mg/kg/day FK506 (tacrolimus), a potent immunosuppressant with neurotrophic properties. Nerve regeneration was evaluated by means of a nociceptive test and a grasping test starting 2 weeks postoperatively. Animals that received bFGF and FK506 showed a significantly faster recovery from injury than did the control group. Morphometric analysis at 3 months showed no difference between the two groups in total number of axonal fibers, fiber diameter, fiber density, and myelin:axon ratio. We conclude that the combination of bFGF and low dose FK506 enhances nerve healing in this animal model by accelerating early regrowth but has no effect on the final outcome. PMID- 12731732 TI - Successful reversal of steal syndrome following creation of arteriovenous fistula by banding with a ringed Gore-Tex cuff: a new technique. AB - Steal syndrome, especially in elderly patients with peripheral vascular disease, is a serious complication following creation of an arteriovenous fistula (AVF) that, if neglected, can lead to amputation. The classic maneuver to deal with the steal syndrome is ligating the AVF and performing another procedure to gain dialysis access. We describe a simple technique of effectively reversing the steal syndrome by banding the vein of the AVF with a ringed Gore-Tex cuff that salvages the AVF and allows its immediate use for dialysis. PMID- 12731733 TI - Post-graduate training in plastic surgery at the department of Professor Ivo Pitanguy: a report. AB - In 1960, Professor Ivo Pitanguy established a Residency Training Program in Plastic Surgery in Rio de Janeiro, Brazil. Since then, more than 500 surgeons from Brazil and around the world have been trained there. This article outlines the organization and functioning of this program and reports an analysis by the author, a former resident and scholar of the International College of Surgeons. PMID- 12731734 TI - Art and surgery: the Hall of Murals at the International Museum of Surgical Science. PMID- 12731735 TI - The role of laparoscopy in the surgical treatment of endometrial cancer. AB - OBJECTIVES: Endometrial cancer is the most common gynaecological cancer. Surgical treatment has traditionally been done by laparotomy, however the laparoscopic approach has gained wider acceptance by gynecologic surgeons. Whether in combination with laparoscopic-assisted or laparoscopic hysterectomy, laparoscopic staging, including lymph-node dissection, is a major component in the treatment of patients with early endometrial cancer. It remains to be proven if these techniques are associated with the greatest benefit. METHODS: Substantial recent studies on the topic of surgical laparoscopic treatment of endometrial cancer were identified from Medline. RESULTS AND DISCUSSION: Laparoscopically assisted surgical staging (LASS) has been reported in several case series totaling more than 600 cases. CONCLUSION: The report illustrates that laparoscopically assisted surgical staging of endometrial cancer is safe as an open procedure. The laparoscopic approach may also be considered for endometrial malignancy which typically occurs in obese and elderly high-risk women. PMID- 12731736 TI - The association of minimal and mild endometriosis without adhesions and infertility with therapeutic strategies. AB - INTRODUCTION: Mild endometriosis may be present in fertile or infertile women. When present in infertile women it could be merely an innocent bystander, and some other problem could be causing the difficulty in conceiving, or it may in some way be directly responsible for the infertility problem. Sometimes to achieve a pregnancy, only these other infertility factors need to be treated with no specific treatment for the endometriosis per se. However there are some data suggesting that sometimes treating the endometriosis surgically may be helpful. METHODS: The pregnancy outcome in women with probable endometriosis vs those without this entity (based on serum CA-125 levels) was compared with treatment rendered only to correcting ovulatory defects with no specific treatment rendered to the endometriotic lesions during the first six months of therapy. Another study evaluated the efficacy of laparoscopic removal of endometriosis vs leaving the lesions untouched on pregnancy outcome in women who failed to conceive after at least eight months of all infertility factors corrected. RESULTS: No difference in pregnancy outcome was found in women with probable endometriosis vs none after six months of correcting ovulation defects. However, for the minority not conceiving after such therapy removing the endometriosis surgically significantly improved fertility rates in the next eight months. CONCLUSIONS: The probable presence of endometriosis based on symptoms, signs, or serologic evidence should prompt careful evaluation and treatment of subtle ovulatory problems, e.g., luteal phase defects and luteinized unruptured follicle syndrome. Therapeutic strategies for those women failing to conceive after 6-8 months of conservative therapy could be laparoscopic removal of observed endometriotic implants or consideration of in vitro fertilization. PMID- 12731737 TI - Cervical carcinoma and pregnancy. AB - Abortion and conization of the cervix is the treatment of choice for patients with preinvasive cervical carcinoma combined with first-term pregnancy. With the second and third term, the pregnancy is led to delivery and a secondary examination is carried out. In case of preienvasive carcinoma only conization of the cervix is performed. In late-term pregnancy the surgical operation starts with a cesarean section. A combination of cervical carcinoma and pregnancy was observed in 31 (44%) of 6,890 patients admitted to the Gynecological Clinic of the National Cancer Center from 1964-2001. Of 1,911 patients with radical hysterectomy (Stage I--58.4%, Stage II--21.8%, Stage III--20%) a combination of cervical carcinoma and pregnancy was diagnosed in 31 women [Stage I--23 (74.2%), Stage II--4 (12.9%), Stage III--4 (12.9%)]; 93.5% of the patients had a first or second term pregnancy. Five-year survival of the patients with surgery only was 83.3%, while with combined therapy--60%. Twenty-nine percent of the patients were 30 and younger. Pregnancy contributed to early manifestation of cervical cancer and did not favor the aggression of malignant tumor growth. The five-year survival rate of patients without staging and those combined with pregnancy was 72.7%; five-year survival rate of patients with early pregnancy was worse compared to those with second or third term pregnancies. Pregnancy is not a contraindication for performance of radical hysterectomy. PMID- 12731738 TI - The effectiveness of hysteroscopic polypectomy in cases of female infertility. AB - The authors, based on their own material, evaluated the value of hysteroscopic treatment of endometrial polyps in a group of infertile women. Polypectomy was performed in 25 patients using either endoscopic microscissors or electric loop. Follow-up hysteroscopy was performed after two months as an integral part of the treatment. All the patients were observed for 12 months. The primary intrauterine investigation was complete in all patients without regard to kind of instrument which was confirmed by second-look hysteroscopy. About 80% of the patients who underwent surgery conceived. Restoration of reproductive ability did not depend on the size of the removed lesion. PMID- 12731739 TI - Endoscopic treatment of intrauterine adhesions. AB - PURPOSE OF INVESTIGATION: The authors investigated the validity of hysteroscopy as a diagnostic and operative technique in cases of intrauterine adhesions. METHODS: 6,680 hysteroscopies were carried out between June 1987 and December 2000 with 201 patients presenting intrauterine adhesions. Only 75 patients underwent resection of the adhesions by hysteroscopy with 94.6% of functional restoration and 93.3% of anatomic resolution. RESULTS: At 2-month follow-up the uterine cavity was completely regular in 70 cases, while in four cases a second surgical treatment was necessary. CONCLUSION: Good anatomic results are obtained after hysteroscopic treatment of intrauterine adhesions, and pregnancy rates vary from 28.7 to 53.6%. PMID- 12731740 TI - Removal of endometrial polyps by use of grasping forceps and curettage after diagnostic hysteroscopy. AB - PURPOSE OF INVESTIGATION: To determine the therapeutic efficacy of the use of gall stone forceps and curettage in endometrial polyps removal, after their detection with diagnostic hysteroscopy. METHODS: From 1997 to 2001, we conducted a prospective study in 53 patients who presented at our department for menstrual disorders, infertility problems or postmenopausal bleeding and in whom endometrial polyps were detected by hysteroscopy. All patients received general anesthesia and after hysteroscopic detection of the polyps' location, their removal was attempted by use of Desjardins gall stone forceps and curettage. Immediately after the procedure, a second hysteroscopy was performed in order to detect remnants of the polyps. RESULTS: Fifty patients presented with only one polyp, two with two polyps and one with three polyps. The mean diameter of the polyps ranged from 0.5 to 3 cm. The hysteroscopic appearance of all polyps was not suggestive of malignancy. During the second hysteroscopy we found parts or whole polyps in five and two cases, respectively, accounting for a therapeutic success of 86.8%. The hospitalization time for all patients was 24 hours and occurred no intraoperative or postoperative complications. CONCLUSION: Our method seems to be safe, with low cost and sufficient therapeutic outcome and could be used in hospitals with availability of diagnostic hysteroscopy only. PMID- 12731741 TI - Trace elements and vitamin levels in menopausal women receiving hormone replacement therapy. AB - Trace elements are extremely important in human metabolism, growth, and tissue repair. The risk of nutritional disturbances, in particular, vitamin and trace element deficiencies, are striking during menopause. The aim of this longitudinal study was to evaluate the effect of estrogen treatment on serum levels of copper, zinc, magnesium, calcium, vitamin E, and vitamin A in menopausal women. Thirty eight menopausal women were included in the study, and were administered a continuous hormone replacement therapy (HRT) of 0.625 mg conjugated equine estrogen (CEE) and 2.5 mg medroxyprogesterone acetate (MPA). Blood samples were obtained before and six months after HRT. There was a statistically significant difference between levels of serum copper, zinc, magnesium, calcium, vitamin E and vitamin A before and after HRT (p < 0.001). In conclusion we observed a beneficial effect of HRT on serum levels of trace elements, vitamin A, and vitamin E in addition to the well known other benefits. PMID- 12731742 TI - The association of minimal and mild endometriosis without adhesions and infertility with therapeutic strategies. AB - INTRODUCTION: Mild endometriosis may be present in fertile or infertile women. When present in infertile women it could be merely an innocent bystander, and some other problem is causing the difficulty in conceiving, or it may in some way be directly responsible for the infertility problem. Sometimes to achieve a pregnancy, only these other infertility factors need to be treated with no specific treatment for the endometriosis per se. However there are some data suggesting that sometimes treating the endometriosis surgically may be helpful. METHODS: The pregnancy outcome in women with probable endometriosis vs those without this entity (based on serum CA-125 levels) was compared with treatment rendered only to correcting ovulatory defects with no specific treatment rendered to the endometriotic lesions during the first six months of therapy. Another study evaluated the efficacy of laparoscopic removal of endometriosis vs leaving the lesions untouched on pregnancy outcome in women who failed to conceive after at least eight months of all infertility factors corrected. RESULTS: No differences in pregnancy outcome were found in women with probable endometriosis vs those without after six months of correcting ovulation defects. However, for the minority who did not conceive after such therapy, removing the endometriosis surgically significantly improved fertility rates in the next eight months. CONCLUSIONS: The probable presence of endometriosis based on symptoms, signs, or serologic evidence should prompt careful evaluation and treatment of subtle ovulatory problems, e.g., luteal phase defects and luteinized unruptured follicle syndrome. Therapeutic strategies for those women failing to conceive after six to eight months of conservative therapy could be laparoscopic removal of observed endometriotic implants or consideration of in vitro fertilization. PMID- 12731743 TI - Introducing colposcopy and vulvovaginoscopy as routine examinations for victims of sexual assault. AB - Victims of sexual assault require appropriate care, follow-up and information regarding their legal rights. Clinicians are faced with the challenging responsibility of identifying victims and providing effective interventive and preventive counselling. The most pressing medical task is to confirm the assault and to undertake correct documentation and exhibition of biological traces. Performing colposcopy and vulvovaginoscopy does not allow us to diagnose a sexual assault trauma, but it can help us to identify those microscopic lesions (due to the enhanced visualization and the higher resolution under which the genital areas are examined) that may not be seen during a normal clinical examination. The colposcopic and vulvovaginoscopic examination starts from the vulvar region looking for superficial lacerations and ecchymosis; the labia majora and minor are examined scrupulously, then the posterior forchette, the perineum and the hymen where it is possible to report microulcerations, contusions and even possible scars due to a precedent defloration. Recent advances in clinical forensic medicine show that trained examiners using colposcopy obtain evidence of genital trauma in 87% to 92% of rape victims. Colposcopy and vulvovaginoscopy must be performed within 48 hours from the sexual assault, because most of the lesions heal rapidly. Colposcopy and vulvovaginoscopy may be seen as a stressful invasion of a woman who is already vulnerable and at risk of the rape trauma syndrome. Prior information about colposcopy may reduce the level of anxiety experienced by many women undergoing this procedure. Incorporating colposcopy and vulvovaginoscopy into the routine assessment of sexual assault victims could be a valid way of identifying genital injuries; moreover the medical report will be more detailed and precise. PMID- 12731745 TI - The adolescent and the gynecologist: our experience. AB - To evaluate adolescents' knowledge about sex, we conducted a study divided in two parts: 1) Distribution of a questionnaire; 2) Impromptu conversation with adolescents. The sample consisted of 205 questionnaires and 115 talks with students at a high school. The gathered data let us deduce that the relationship with parents was regarded as pleasant and good. As was expected the whole emotional sphere was related to friends. In order to understand which structures can help adolescents our data revealed that the mother holds first place, followed by the school and by friends. Sex education during adolescence should be carried out by other institutions so that a relationship free from conflicts can be established. In this way the figure of the expert takes shape--someone who can communicate within the structure of the school and the family and beyond--the limits of the information given by friends. PMID- 12731744 TI - Influence of angiotensin converting enzyme inhibitors on stable myocardial ischemia in menopausal cardiac patients. AB - PURPOSE OF INVESTIGATION: OBJECTIVE: The aim of the study was to investigate the influence of angiotensin converting enzyme inhibitor (Captopril) on stable demand ischemia in menopausal cardiac female patients. METHODS: In a prospective non randomized evaluation of the effect of angiotensin converting enzymes inhibitors (ACE-I) (Captopril) on demand myocardial ischemia, 16 normotensive menopause female patients, mean age of 52 years with stable angina and known coronary artery disease (CAD) but normal left ventricular function, underwent a treadmill exercise test (Bruce Protocol), at baseline (T1) and one week following (50-75 mg) a daily dose of Captopril (CAPT2). Onset of symptoms, duration of exercise, magnitude of peak ST depression and homodynamic parameters were monitored. RESULTS: Captopril significantly increased the duration of exercise from (467 +/- 169 to 536 +/- 145 seconds, (p value < 0.02), but with increased peak of ST segment depression (from -1.4 +/- 0.6 mm to -1.93 +/- 1.2 mm, p value < 0.15). The double product remained unchanged (251 x 103 +/- 55 in T1 and 248 x 103 +/- 55 in CAPT2; the p value was < 0.8. All adverse effects on the treadmill were noted. CONCLUSION: Although captopril tends to significantly effect prolongation of exercise time, there is no amelioration of the markers of ischemia. PMID- 12731746 TI - Body weight recovery in intrauterine growth-retarded rats treated with growth hormone. AB - The aim of this work was to analyze the action of growth hormone (GH) on postnatal body weight recovery in intrauterine growth-retarded (IUGR) rats. Wistar rats were assigned to three groups: 1) control; 2) IUGR and 3) sham operated. Uterine vessels of dams in the IUGR group were partially bent on the 14th day of pregnancy. At weaning, some IUGR pups were randomly selected and injected with GH (3 mg/kg/day), up to the 60th day. A standard diet ad libitum was available to mothers and offspring. The animals were weighed and food intake was recorded weekly. The weight gained velocity and relative food intake (RFI) was calculated. IUGR animals showed significant lower body weights than the control group. GH treatment allowed body weight recovery in IUGR rats. In females, body weight increased 14 days before males, and the former had greater RFI values. In conclusion, our results indicated differences in sexual responses to GH treatment. There is a need for more research on the mechanisms involved in that sexual difference. PMID- 12731747 TI - The effect of smoking and caffeine on the fetus and placenta in pregnancy. AB - The aim of this study was to determine the effects of tobacco use and consumption of caffeine in pregnancy on the fetus and placenta by measuring the body weights, head circumferences, and lengths of newborns, and also weights and diameters of placentas. In this prospective study, two main groups were chosen for the study: Group I: A total of 63 pregnant non-smokers; were separated into two subgroups according to their daily caffeine intake; less than 300 mg (Ia) (n = 44), and more than 300 mg (Ib) (n = 19). Group II: 60 pregnant smokers were also separated into two subgroups; daily caffeine intake less than 300 mg (IIa) (n = 43), and more than 300 mg (IIb) (n = 17). The newborns and placentas of both groups were examined. The body weights, lengths, and head circumferences of newborns and also weights and diameters of placentas were measured. The pregnant non-smokers consuming caffeine more than 300 mg/day had statistically significant lower weights of newborns and placentas (p < 0.05). However, there was no significant difference between groups according to the lengths, head circumferences of newborns and diameters of placentas. There were significantly lower body weights of newborns and placentas in pregnant smokers (p < 0.05). There was no difference according to the diameters of placentas, and lengths and head circumferences of newborns in either group. In conclusion, it is suggested that smoking in pregnancy should be prevented both for the health of newborns and mothers, and also caffeine including beverages like tea and coffee should be limited in pregnancy. PMID- 12731748 TI - Early ultrasonographic diagnosis of unruptured interstitial pregnancy: a case report and review of the literature. AB - Interstitial ectopic pregnancy occurs when the fertilized ovum implants in the interstitial portion of the fallopian tube, where it transverses the wall of the uterus from the cornua to the corner of the uterine cavity. Interstitial ectopic pregnancy is an infrequent type of ectopic pregnancy, occurring in 2% to 4% of all tubal pregnancies. It is a life-threatening condition because myometrium rupture tends to occur in the second trimester of pregnancy resulting in profuse bleeding. This paper concerns the ultrasonographic diagnosis of a 33-year-old, gravida 4, para 3 woman with unruptured interstitial pregnancy, diagnosed in the 7th week of pregnancy. Awareness and suspicion of such pregnancy are required for its early detection. A combination of the sensitive beta-hCG assays, a single serum progesterone measurement and transvaginal ultrasonography are the principle tools used to make the diagnosis. The diagnostic difficulties are also discussed. PMID- 12731749 TI - Severe hypoplasia of the right femur, ipsilateral agenesia of the fibula and twisted right foot in a 24-week-old fetus with proximal femoral focal deficiency (PFFD). AB - A case of a 24-week-old fetus of non consanguineous parents with an ultrasonographic diagnosis of a short right femur, ipsilateral agenesia of the fibula and a twisted right foot is described. Cordocentesis revealed a normal 46XY karyotype. The parents were informed on treatment options and after psychological counselling they decided to undergo an abortion. Post mortem examination confirmed the diagnosis of a severe right limb malformation. PMID- 12731750 TI - Unusual case of cervical pregnancy after curettage for a presumptive diagnosis of intrauterine blighted ovum. AB - A case of cervical pregnancy after curettage for presumptive intrauterine blighted ovum is presented. The woman was successfully treated by vacuum evacuation and curettage. A 29-year-old woman, gravida 2, nulliparous, was admitted to our department at ten weeks and two days of gestation after a diagnosis of cervical pregnancy. She had been treated by curettage five days before for an initial diagnosis of intrauterine blighted ovum. Ultrasound scan examination revealed a gestational sac without foetus in the cervix four days after the first curettage. Vacuum evacuation and curettage of the cervical canal were performed and a Foley catheter was also inserted and left in place for three days. The patient was discharged in good condition on the fourth postoperative day. PMID- 12731751 TI - Placenta percreta: report of two cases and review of the literature. AB - Placenta percreta is a serious complication of pregnancy. Two cases of placenta percreta confirmed histologically were treated by supravaginal hysterectomy. Case 1: A case of uterine rupture secondary to placenta percreta was diagnosed in a 29 year-old term primigravida during an elective abdominal delivery of a healthy fetus. Spontaneous rupture of the primigravid uterus due to placenta percreta without a history of trauma or infection is a very rare occurrence. Case 2: A 33 year-old previously healthy G4P2 woman was admitted at 29 weeks of gestation with acute abdominal pain and hemorrhagic shock. There was a history of one induced abortion and two cesarean section deliveries. A review of risk factors, diagnostic tools and treatment possibilities are given. PMID- 12731752 TI - Genotypic analysis methods for detection of drug resistance mutations in the HIV 1 proteinase and reverse transcriptase genes. AB - Understanding the basis of human immunodeficiency virus (HIV) drug resistance represents a key requirement for individualized HIV patient care. The genotypic data generated to date have already provided significant insight. However, it is clear that the relationship between genotype, phenotype and clinical outcome is complex and still poorly defined. In this review, we describe methods currently available to obtain genotypic data for the HIV-1 proteinase and reverse transcriptase genes. Different sample preparation strategies and DNA sequencing methods are discussed dividing the latter into two categories, those that give sequence information at specific positions and those that provide continuous sequence data for a particular region. In addition, we also address some of the broad biological and technical issues, which must be considered when interpreting the results of these tests and describe the advantages and disadvantages of individual methods. PMID- 12731753 TI - Safety and efficacy of once daily intranasal zanamivir in preventing experimental human influenza A infection. AB - Zanamivir, a potent inhibitor of influenza A and B virus neuraminidases, is protective against experimental human influenza when given intranasally twice daily. We conducted two studies to assess the pharmacokinetics and protective efficacy of a reduced frequency dosing regimen of topical zanamivir. In the first study, 36 uninfected volunteers received a single dose of zanamivir by intranasal spray (6.4 mg), intranasal drops (16 mg) or dry powder oral inhalation (10 mg). At 4 h, median nasal wash concentrations were 50-fold higher after intranasal dosing than after inhalation. Substantial levels (spray group, median 4,596 ng/ml; drop group, 1,239 ng/ml) were detected in nasal wash 48 h after intranasal dosing. In the double-blinded efficacy study, 47 sero-susceptible volunteers were randomized to receive either placebo or zanamivir intranasal spray (6.4 mg). Among the 43 subjects evaluated, decreases in viral shedding occurred in the group receiving one dose of zanamivir 4 h prior to inoculation, whereas no significant benefit was observed in those receiving a single dose 48 h prior to challenge. In the group given three daily doses, reductions were seen in viral shedding and infection. In the two regimens providing zanamivir 4 h prior to inoculation, significant reductions in nasal mucus weight were observed. Decreases in total symptom scores and the incidence of upper respiratory illness also occurred, but they did not reach statistical significance. The efficacy of a single dose of zanamivir given 4 h prior to inoculation supports the hypothesis that influenza virus neuraminidase is essential for initial virus spread through respiratory secretions. These findings indicate that once daily dosing of zanamivir is protective against experimental influenza A infection. PMID- 12731754 TI - Antiretroviral effect of two different dose regimens of ritonavir and saquinavir on HIV-infected adults in a population-based setting. AB - OBJECTIVE: To characterize the antiviral effect and tolerability of higher dose (HD, 600 mg two times daily) and lower dose (LD, 400 mg two times daily) combination regimens of ritonavir and saquinavir in British Columbia (BC), Canada. DESIGN: Intent-to-treat analysis with suppression of plasma viral load to levels below 500 copies/ml as the main outcome measure. PATIENTS: Adult human immunodeficiency virus (HIV)-positive individuals in the province of British Columbia prescribed ritonavir and saquinavir in combination between 1 September 1996 and 30 June 1997, with a minimum of two plasma viral loads, one at baseline and one after the initiation of therapy. RESULTS: A total of 84 participants [27 HD (32%) and 57 LD (68%)] were prescribed ritonavir and saquinavir. There was no difference at baseline in the two groups with respect to age (P=0.466), CD4 cell count (P=0.373) and baseline plasma viral load (P=0.656). However, LD were more likely to have had prior protease experience than HD participants (65 versus 40%, P=0.037). The median follow-up time was 9 months. A total of 44 (52%) subjects demonstrated a decrease in plasma viral load to levels <500 copies/ml. After adjusting for length of follow-up, baseline CD4 cell count and prior AIDS diagnosis, HD participants were as likely to be suppressed to <500 copies/ml as LD individuals (P=0.760). HD participants did report more adverse effects (P=0.042) than LD subjects. CONCLUSION: Our results provide confirmatory evidence that lower doses of ritonavir and saquinavir in combination are better tolerated and as effective as the standard doses of these drugs. This response, however, is seriously compromised by prior exposure to protease inhibitors. PMID- 12731755 TI - Percentage of adherence correlates with the risk of protease inhibitor (PI) treatment failure in HIV-infected patients. AB - OBJECTIVES: To determine the effect of adherence on the rate of protease inhibitor (PI) treatment failure among human immunodeficiency virus (HIV) infected patients. METHODS: A prospective study of a cohort of 282 patients who initiated PI therapy from March 1996 to December 1997. Adherence was quantified as the percentage of prescribed doses reportedly taken and treatment failure was defined as HIV RNA levels above 200 copies/ml after 1 year on therapy. RESULTS: Overall, 190 patients (67%) missed prescribed doses. However, mean percentage of doses taken was 91% (range, 21-100%). Demographic, virological and immunological characteristics could not predict adherence outcomes. The causes of non-adherence included intolerance or side effects (35%), complexity of treatment (23%), or recurrence in active drug abuse (17%), whereas abandonment owing to HIV-related disease was uncommon (6%). A degree of adherence above 90% correlated significantly with viral suppression [relative risk (RR) 1.69; 95% confidence interval (CI) 1.1-2.56; P<0.01]. In a multivariate analysis, a lower degree of adherence (RR, 0.96; P=0.006), a higher HIV viral load (RR, 2.03; P=0.0001), prior antiretroviral therapy (RR, 2.5; P=0.01), and use of saquinavir-hard gel capsules (saquinavir-HGC) (RR, 1.77; P=0.03) were strongly associated with treatment failure. CONCLUSION: The percentage of adherence and initial HIV viral load are the most important determinants of virological response to PI therapy and non-adherence is related to treatment-related factors in the majority of cases. PMID- 12731756 TI - Lipid evaluation in HIV-1-positive patients treated with protease inhibitors. AB - There is accumulating evidence that human immunodeficiency virus type 1 (HIV-1) protease inhibitors (PIs) can induce hyperlipidaemia. To evaluate the frequency and type of hyperlipidaemia in PI-treated patients, 98 outpatients were prospectively analysed for their lipoprotein characteristics at the Medizinische Hochschule in Hannover, Germany. Fifty-seven percent of the patients studied presented with hyperlipidaemia. Both hypertrigylceridaemia (type IV and V hyperlipoproteinaemia, 33%) and hypercholesterolaemia (type IIa hyperlipoproteinaemia, 6%) were detectable. The remaining 18% had a type IIb hyperlipoproteinaemia. Increased lipid levels were highly statistically significant compared to a control group of PI-naive HIV-1-infected patients [low density lipoprotein (LDL) 146 mg/dl (range, 53-274 mg/dl) versus 105 mg/dl (range, 22-188 mg/dl; P=0.0006); very-low-density lipoprotein (VLDL) 35.5 mg/dl (5-253 mg/dl) versus 18 mg/dl (range, 3-94 mg/dl; P=0.0002)]. All PIs used (saquinavir, indinavir, nelfinavir and ritonavir) were associated with this variable form of hyperlipidaemia according to the Fredrickson classification. There was no significant correlation of any determined lipid value with the duration of treatment. A higher frequency of the apolipoprotein E2 allele and E4 allele was observed in the hyperlipidaemic subjects. Patients with excessive hypertriglyceridaemia showed a reduced lipoprotein lipase activity. Lipodystrophy was observed especially in hyperlipidaemic patients and to a lesser extent in normolipidaemic subjects. The frequency of hyperlipidaemic risk factors was surprisingly high in the group studied, which in turn may explain the proposed increased risk of atherogenesis in HIV-1 PI-treated patients. Therefore, PI treated subjects should also be evaluated for their lipoprotein pattern, which may require antihyperlipidaemic interventions. PMID- 12731757 TI - Selection and characterization of HIV-1 variants resistant to the (+) and (-) enantiomers of 2'-deoxy-3'-oxa-4'-thiocytidine (dOTC). AB - Human immunodeficiency virus type 1 (HIV-1) variants were selected for resistance against the (+) and (-) enantiomers of a novel nucleoside analogue, 2'-deoxy-3' oxa-4'-thiocytidine (dOTC), using the infectious molecular clone HXB2D grown in the MT-4 line of human T cells. The variants selected with (+) dOTC were approximately 6-7-fold less sensitive than wild-type virus to this drug. Cloning and sequencing of the complete reverse transcriptase (RT) coding region of these variants identified the M1841 mutation and further selection with virus containing the M1841 substitution led to the appearance of an M184V mutation. In contrast, selection experiments performed with (-) dOTC yielded variants capable of growing in drug concentrations as high as 100 microM, but phenotypic analysis of these viruses revealed near wild-type 50% inhibitory concentration (IC50) values for this compound. Site-directed mutagenesis experiments in which the M1841 and M184V mutations were introduced into HXB2D confirmed the importance of these mutations when viruses were grown in MT4 cells. However, wild-type IC50 values in regard to both (-) and (+) dOTC were obtained when these recombinant viruses were grown in cord blood mononuclear cells (CBMC). Clinical isolates of HIV-1 resistant to lamivudine and containing the M184V substitution also displayed low-level resistance to both (-) and (+) dOTC when grown in CBMC. Finally, cell-free RT assays were performed in the presence of either (-) dOTC triphosphate, (+) dOTC triphosphate, or the triphosphate of a racemic mixture of (+) and (-) dOTC with wild-type and mutated M184V-containing recombinant RT. The data demonstrate chain termination effects of these compounds with regard to both wild-type and mutated enzyme and that the latter was approximately twofold less sensitive than the former to these drugs. PMID- 12731759 TI - Anti-HIV-1 intermittent drug intensification in HIV-1-infected patients naive for antiretrovirals. PMID- 12731760 TI - Opening speech of the 12th Stockholm Water Symposium. PMID- 12731758 TI - Evolution of genotypic resistance to nucleoside analogues in patients receiving protease inhibitor-containing regimens. AB - The prevalence of genotypic resistance to nucleoside analogues (NA) was examined using a line probe assay (LiPA, Innogenetics, Spain) and a point mutation assay to test for codon 151 polymorphism in plasma from 34 individuals who had been exposed to NA for longer than 1 year. The testing was repeated in the same population after 6 months of being on a new potent protease inhibitor (PI) containing antiretroviral combination. Only nine (47%) of the 19 patients initially carrying the codon 41 mutation restored zidovudine wild-type (WT) virus population. Similarly, eight (33%) out of 24 carrying the codon 215 mutation restored the wild-type variant. Two subjects carrying codon 74 didanosine mutation reverted to wild-type genotype, as well as two (18%) out of 11 harbouring the codon 184 lamivudine-resistant variant. To conclude, the extent to which drug recycling might be of benefit in subjects showing a restoration of genotypic sensitivity to former drugs needs to be explored. PMID- 12731761 TI - Opening address of the 12th Stockholm Water Symposium. PMID- 12731762 TI - International freshwater policy from Bonn to Johannesburg and beyond. PMID- 12731763 TI - Balancing priorities: the role of industry in water resource management. PMID- 12731764 TI - Summary and conclusions of the 2002 Stockholm Water Symposium. PMID- 12731765 TI - 2002 Stockholm Statement: urgent action needed for water security. PMID- 12731766 TI - Balancing competing water uses--a necessity for sustainable development. AB - The UNEP Water Policy and Strategy emphasises the need for environmentally sustainable water use as a key to sustainable development, including poverty reduction. The development of UNEP programmes over the years has produced an integrated, comprehensive and dynamic approach to competing water uses. PMID- 12731767 TI - Hydrologic dynamics and ecosystem structure. AB - Ecohydrology is the science that studies the mutual interaction between the hydrological cycle and ecosystems. Such an interaction is especially intense in water-controlled ecosystems, where water may be a limiting factor, not only because of its scarcity, but also because of its intermittent and unpredictable appearance. Hydrologic dynamics is shown to be a crucial factor for ecological patterns and processes. The probabilistic structure of soil moisture in time and space is presented as the key linkage between soil, climate and vegetation dynamics. Nutrient cycles, vegetation coexistence and plant response to environmental conditions are all intimately linked to the stochastic fluctuation of the hydrologic inputs driving an ecosystem. PMID- 12731768 TI - The role of regulatory agencies in multiple water use. AB - This paper presents the conceptual framework for regulating water use in cases where water is a public good. Public agencies with an independent directive board can successfully manage such a situation. Through a comprehensive system of water permits, charging and control it is possible to effectively manage water resources. Together with the conceptual framework a practical application is presented. The case of the National Water Agency of Brazil--ANA--shows that it is possible to implement modern water resources management concepts in less developed countries. The benefits of this institutional arrangement are demonstrated in the case of managing water conflicts among water users. Two situations are described: the semi-arid water allocation for agricultural use in the Jaraguaribe River Basin in Ceara State and the conflict between the hydropower and navigation sectors in the Parana River Basin. PMID- 12731770 TI - Stakeholder involvement in water management: necessity or luxury? AB - Stakeholder involvement in water management is widely recognized as an important component of the design and implementation of sustainable water management initiatives. Despite this, there remains a deep-rooted resistance to the widespread implementation of programs to prioritize such involvement (as witnessed by, for example, the low priority given to the public involvement element of the European Union Water Framework Directive). This paper addresses the issue of stakeholder involvement by first confronting the fact that it is not a water issue, per se. Such diverse fields as economics, agriculture, public health, pollution prevention, business and education have also identified stakeholder involvement as a difficult but necessary component of successful action in their fields. For the water sector, the issue of stakeholder involvement as either a necessity for sustainable water management, or a luxury to be used to complement traditional approaches, is discussed. PMID- 12731769 TI - Water pricing in England and Wales--institutions and objectives. AB - Water pricing in England and Wales reflects a range of influences. Cost recovery is generallythe primary influence over prices. The privatised public water supply sector dominates water usage. Water abstractions are licensed and allocated mainly on an administrative basis. Charges are levied to recover the water management costs of the licensing authority (the Environment Agency), and are not closely related to the scarcity of water resources, or the environmental impact of abstractions. Public water supplies are subject to price cap regulation. This provides incentives to reduce costs. Public water suppliers are expected to choose an optimal combination of water resource interventions in planning water supplies. The system of regulation has exposed choices. Tariffs for customers reflect company and regulatory concerns to share the burden of cost recovery equitably. Tariff innovation to influence behaviour has been limited by the low extent of metering of household customers. The cost recovery and institutional framework has led to discipline in investment decision-making. PMID- 12731771 TI - Water, food and environment: conflict or dialogue? AB - Increased water conflicts are the result of the rapidly increasing relative water scarcity in many basins to which water institutions have not yet successfully adapted. The Dialogue on Water, Food and Environment is a program of activities that employs processes at international, national, basin and local scales to help build bridges between agriculture and the environment. PMID- 12731772 TI - Ecological economics for water policy and management: a need for major shifts in thinking. AB - Economics cannot be separated from values and ideology. Attempts to influence the political agenda by raising the fundamental issues of paradigm, ideology and institutions in various arenas are crucial. is recognized that problems related to water quality and availability are linked with all kinds of activities and sectors in society. PMID- 12731773 TI - Forward to Johannesburg and hydro-solidarity. AB - The provision of clean drinking water and adequate sanitation is one of the simplest and most effective steps to eradicating poverty. In South Africa a major programme is underway which will ensure all South Africans have a clean water supply within six years, and access to acceptable sanitation within ten years. Total basin management, transcending national boundaries, is essential to ensure the use of water for the common good of all. The term "hydro-solidarity" illustrates water as a catalyst for co-operation. PMID- 12731774 TI - Virtual waterless manufacturing: zero intake, zero emission. AB - Innovative application of a systematic approach to reduce freshwater intake and achieve zero emissions could help in reducing the adverse impact of industrial activity on world water resources. Cleantech is a strategic enviro-management technique to reduce the generation of pollutants in a process at source, through minor process modification, material substitution, improved manufacturing practices or low cost treatment. PMID- 12731775 TI - Beyond the river: the benefits of cooperation on international rivers. AB - Management of international rivers can be a cause of conflict or cooperation between states. Benefits of cooperation include benefits to the ecological river, the economic river, the political river and the catalytic river. The challenges are in balancing the trade-offs between states. PMID- 12731776 TI - Making markets work for clean water. AB - Part of the solution to close the gap to meet the UN Millennium safe water goal lies in the domestic consumer marketplace. Multinational corporations must design products for low income consumers to deliver clean water at a household level. The future of business is linked to improving the lives of developing country consumers and to the improved economic status of those consumers. PMID- 12731777 TI - Workshop 1 (synthesis): water as a catalyst for development. AB - The problem of allocating water resources is a political one, and water management teams now need to include political lobbyists and advisors. Education, at all levels, is a crucial element in any development strategy involving water. PMID- 12731778 TI - Politics and governance at the interface between water and development. AB - Water is not just a technical issue--it always has to be addressed within a complex political context. The water sector professional is generally unskilled in political processes, but political skill and awareness is needed to avoid negative politicization of water. PMID- 12731779 TI - Workshop 2 (synthesis): water pollution abatement within the industrial sector. AB - This workshop aimed at demonstrating and discussing how effective abatement of water pollution can be achieved through introducing cleaner technologies, recycling and reuse of water, and implementing new public policy measures. PMID- 12731780 TI - Micro enterprise initiative in water sector and poverty reduction . AB - The author reports on the Kerala model for water sector development, broadly adopted as a role model for poverty reduction and build up of social capital. It is a community based organisation with its focus on facilitating a stable income to the poor, and composed of a People's Plan Campaign, the Kudumbashree (women based poverty eradication programme), with grassroot level neighbourhood groups, federated into an area development society. It promotes savings and credit channelling, capacity building and entrepreneurship development. Activities include awareness raising on water conservation and hygiene, utilization of student power, promotion of small, cheap and low technology projects that people can understand and undertake (small reservoirs, tanks, rainwater harvesting structures, water meters), as well as microenterprises, and training of women based repair groups. PMID- 12731781 TI - Workshop 3 (synthesis): water, poverty alleviation and social programs. AB - Poverty is a complex issue, which must be understood in a holistic manner. Low and variable income is certainly a key element, but it is far from enough to portray poverty. The various characteristics of poverty and their relative strength are determined through contextually specific circumstances, in terms of history, environmental preconditions, socio-cultural traits, etc. Much of this context is made up of local and national circumstances. The consequences of globalisation must, however, increasingly to be taken into account. At a larger scale, it is also relevant to mention that climate change will have a negative, although largely unpredictable, impact for the people in some parts of the world. For those who are already living on marginal lands or who are exposed to water problems, climate change is likely to create considerable adverse effects. PMID- 12731782 TI - Towards poverty alleviation: the water sector perspectives. AB - Water and poverty interface in many different ways. Sustainable management (i.e. development, allocation and utilization) of water resources is therefore a process-element of sustainable human development. This paper looks into water poverty interfaces as well as into approaches to, and tools of, managing water in such a manner that water sector activities can contribute to alleviation of poverty. It gives a southern perspective and includes a case study of the evolving water management regime in Bangladesh. PMID- 12731783 TI - Water participation for poverty alleviation--the case of Meseta Purepecha, Mexico. AB - The construction of small water reservoirs has been used in an effort to alleviate poverty in Messeta Purepecha region in Mexico. The programme's rationale can be characterised as incentive-based participation, using both local employment and shared risks concepts. The programme so far has been a relative success. However, in the light of poverty alleviation questions have to be raised about the isolated nature of the programme as well as the role of the incentives used. PMID- 12731784 TI - Workshop 4 (synthesis): criteria for priorities between competing water interests in a catchment. AB - Growing demand for water leads to increased competition between water users. A holistic, participatory and decentralized water management approach is promoted to reach a fair allocation mechanism between competing water uses. Domestic political discourse has a strong influence on water policies being developed. PMID- 12731785 TI - Alleviating water scarcity in Northern China: balancing options and policies among Chinese decision-makers. AB - Water scarcity is mostly a man-made problem that increasingly affects people's lives and questions economic output in Northern China. Policy options addressing the serious water shortages in the region include conservation and water management reform, which is unpopular; grain imports and the downsizing of agriculture; derivation of Yangze water; derivation of the Amur and other international rivers. All solutions present major difficulties, whether domestic or international, for the Chinese authorities. While major decisions have not yet been taken, evidence shows major works such as water transfers from the Yangze or water pricing are not only unavoidable, but will not be enough to meet the growing demand for water in the region. PMID- 12731786 TI - The power of the "sanctioned discourse"--a crucial factor in determining water policy. AB - The water relations in the Jordan River Basin are an often-analysed case of decision-making under terms of water scarcity. However, most of these studies fail to address the underlying structures, which are the focal point in the formation of each of the basin parties' water policy. This paper argues that it is necessary to analyse the domestic structures of the parties since the policies pursued in the international arena are likely to be a reflection of the domestic discourse. Particular emphasis is put on how domestic structures affect and create sanctioned discourses and how these in turn affect foreign policy decision making with regards to water. The various dominant discourses in three of the parties in the Jordan River Basin--Jordan, Israel and the Palestine Authority- are analysed. Failing to acknowledge the explanatory power of this line of thinking runs the risk of reaching simplified conclusions, such as--"the policy makers do not understand water issues"--instead of acknowledging that the power of the discourse perhaps gave them little choice. The paper draws on a variety of disciplines such as international relations theory, sociology and science studies. PMID- 12731788 TI - From Rio via the Hague to Johannesburg: the role of multi-stakeholder dialogues. AB - The Stakeholder Forum will contribute a number of partnership initiatives in the areas of food security, freshwater, energy and health. These will demonstrate that stakeholders, working in partnership, can play their roles and meet their responsibilities in delivering sustainable development agreements. PMID- 12731787 TI - Workshop 5 (synthesis): stakeholder participation in decision making- institutional forms. AB - Water problems have a human dimension where the ideas, perceptions, values and life-styles of individuals in their different roles have an impact. A many-sided set of stakeholders is preferable in any decision process, with empowerment of those less able to make their views known. PMID- 12731790 TI - Workshop 6 (synthesis): water pricing. AB - Water shold be costed so that it is valued, and fees should be transparent and contribute to adequate cost recovery with consideration of social, economic and environmental effects. There is a need for targeted subsidies to assist the poor. Water pollution fees for point sources can reduce discharges. PMID- 12731789 TI - Linking stakeholders and decision makers with science in managing the coastal water environment: case studies from urban, industrial and rural subtropical catchments in Australia. AB - The dynamics of stakeholder participation and effective dialogue processes vary markedly between and within catchment areas. Decision support tools and conflict resolution skills are essential in developing consensus in complex and conflicting issues. Undertaking relevant research requires genuine inclusion of all stakeholders at all stages of the process including development and implementation of management plans. Providing a scientifically integrated and participative approach increases the ability to understand the social and economic dimensions. PMID- 12731791 TI - Workshop 7 (synthesis): integrating the water and energy sectors. AB - Integration of the water and energy sectors is a key issue for effective resource management. However, everything that seems to make sense cannot always be easily accomplished. Barriers are ultimately related to political will where a variety of "instruments" is needed to make such coordination effective. PMID- 12731793 TI - Workshop 8 (synthesis): urban dynamics, livable cities and water. AB - A major wave of urbanisation is taking place in small- and medium-sized cities in developing countries. Water governance and integrated approaches are key factors in providing adequate services and maintaining the water resource. Intermediate technologies and practices need to be applied on a much wider scale. PMID- 12731792 TI - Dyadic design interface between energy and agriculture: the case of Pinthali micro hydro system in Nepal. AB - Technology, like society, is heterogeneous. It mirrors the context in which it operates. Micro hydro development in Nepal is a rural energy strategy, which relies on technology and innovation and takes place in a specific social context. In designing this energy strategy, both technology and its social context, therefore, need to be considered seriously. In technical design processes, the interplay between the content (technology) and the context (society) needs to be considered, as the outcome will affect the people. For example, the content- micro hydro system--in the domain of the context--agriculture--provides an arena for an integrated water control system. Thus, it is possible to control water for two purposes: to produce power and to provide irrigation. The end product will be "energy" as a "consumptive" output and improved food security as a "productive" output of water. Therefore, within a sociotechnical framework, energy and irrigation become constitutive outputs of the sacrosanct "water". Thus, the metaphor of power--the "sociotechnical code" of "content" and "context"--can be used with the term "agro-anergy" in the design process of micro hydro systems. Evidence suggests that this interaction can lead to a transformed water use system for both productive and consumptive output for the benefit of rural communities. PMID- 12731794 TI - Towards a typology of water-related conflicts in the urban environment. AB - Rapid urban population growth is occurring, particularly in developing countries, and has led to problems of providing adequate water supply and sanitation. Two dominant policy recommendations have been put forward--raise fees to existing users or increase the role of the private sector in the provision of water and sanitation services. A number of civil disturbances and riots have occurred in response to such proposals. More pragmatic policy initiatives are required to prevent and solve water-related conflicts, as well as new institutional structures to better handle competing and conflicting water demands. PMID- 12731795 TI - Summary and conclusions from the SIWI seminar for young water professionals water and sustainable development--how to ensure development without compromising sustainability? AB - There is a need to create a balance between development and conservation in order to find a way to mitigate the conflicting interest of water for society, the environment and the economy. Apart from finding a solution to this there is a need to get the mesaage across to the decision makers. How do we make good ideas permeate policy and translate into concrete programs? The Young Water Professionals gave their view through presentations and discussions. It was argued that the answer was not to be found only in environmental science but also in the political and social arena. It was argued that the sanctioned discourse is a powerful force in water allocation and management. How can a balance be struck? Many argued for a unique design of a policy for the whole catchment, acknowledging ecology and existing institutions. Furthermore, many argued in favor of building on existing institutions and steward groups for sustainability and increase their adaptive capacity. PMID- 12731796 TI - The contemporary psychoanalyst at work. PMID- 12731797 TI - Telling stories. AB - Patients need to tell their stories. One of our primary tasks as analysts is to help patients tell their stories and own them. The freedom of mind to think, to feel, and to know are dependent on the ongoing capacity for storytelling. The analyst's stance plays a major role in the development of the analysand's storytelling capacities. PMID- 12731798 TI - The fate of the dream in contemporary psychoanalysis. AB - Freud's metapsychology of dream formation has implicitly been discarded, as indicated in a brief review of trends in psychoanalytic thinking about dreams, with a focus on the relationship of the dream process to ego capacities. The current bias toward exclusive emphasis on the exploration of the analytic relationship and the transference has evolved at the expense of classical, in depth dream interpretation, and, by extension, at the expense of strengthening the patient's capacity for self-inquiry. This trend is shown to be especially evident in the treatment of borderline patients, who today are believed by many analysts to misuse the dream in the analytic situation. An extended clinical example of a borderline patient with whom an unmodified Freudian associative technique of dream interpretation is used with good outcome illustrates the author's contrary conviction. In clinical practice, we should neglect neither the uniqueness of the dream as a central intrapsychic event nor the Freudian art of total dream analysis. PMID- 12731799 TI - The analyst's desire and the problem of narcissistic resistances. AB - The ways in which the analyst's desire for particular experiences with patients is inevitable and often leads to narcissistically based resistances are considered. Five propositions are examined: (1) that the analyst cannot help but have desires and want them recognized by the analysand; (2) that these desires frequently underwrite the analyst's theoretical beliefs and technical interventions; (3) that narcissistic desires and their influence are ubiquitous among practicing analysts; (4) that the patient is often on the lookout for the analyst's various agendas; and (5) that the patient often hopes the analyst will put his or her desire aside and listen so the patient can further his or her own interests. Lacan's concept of the "dual relation" is central to this discussion. The neo-Kleinian position on narcissistic resistances is explored, as is the idea of the "analytic third" as a potential solution to the problem they pose. An extended case description illustrates the main points. PMID- 12731800 TI - Analysis and psychotherapy by telephone: twenty years of clinical experience. AB - Psychoanalysis via telephone is becoming increasingly prevalent while remaining an area of comparatively little study. The author's early telephone treatment of a series of patients living some distance away or engaged in business travel, and his subsequent telephone treatment of nine analytic and five psychotherapy patients following his own geographic move, are discussed in detail. The mechanics of beginning and carrying out such treatment are examined. The theoretical implications of the shift to the telephone and the ambivalence with which it is often met by clinicians are also explored. The role of nonverbal communication in both in-person and telephone analysis is considered, as is the concept of the analytic office as a literal space and a psychological container. Suggestions for future research are advanced. PMID- 12731801 TI - The holding function of theory. AB - As multiple theoretical models contend on the American analytic scene, the holding function of theory emerges as a unifying theme. In addition to supplying an intellectual superstructure for the working analyst, theory provides a psychological presence--a sense of conviction, affective stability, reassurance, and self-esteem--that makes effective analytic work possible from the analyst's side. Ideological passions and differences arise from the vital need for the holding function in an intense and inchoate engagement like psychoanalysis. To show how adherents of different models use theory in practice, three clinical cases are reviewed, one from Betty Joseph, one from Lewis Aron, and one from the author. The last example reconstructs the analyst's subjective experience of treatment both in the selected hours and in terms of the analyst's preconscious use of theory. PMID- 12731802 TI - Silent thoughts, spoken wishes: when candidate experience of the supervisor converges with patient fantasies. AB - How and why a candidate's private experience of two supervisors emerged in patients' fantasies about them is explored. Four issues are examined in light of two control cases: (1) Patients divide, rather than split, the transference between supervisor and candidate, experiencing both ambivalently. (2) Even a patient with no knowledge of the supervisor's identity may have a fantasy of the supervisor that is congruent with the candidate's experience of the supervisor. (3) When new professional traits emerge in the candidate as he or she identifies with his or her mentor, the patient may attribute them to the invisible person in the room--the supervisor; the patient may intuit and be influenced by the candidate's feelings about the supervisor as well. (4) A patient's fantasies about the supervisor may reflect parallel process in reverse, whereby the patient discerns what is going on between supervisor and candidate through his or her treatment, just as the supervisor reads what is going on between patient and candidate through the candidate's reporting of the treatment. Because the trio is the truth of the training case, it seems fitting and empowering to acknowledge and analyze the role of the supervisor in the patient's mind. PMID- 12731803 TI - Being a candidate: its impact on analytic process. AB - Despite its crucial importance to psychoanalytic training, the control analysis has received surprisingly little scrutiny by psychoanalytic writers. How the candidate's training influences analytic process--a critical feature of the control analysis--is examined. In keeping with previous contributions, it is found that the candidate's training experience does indeed influence analytic process. It is argued, however, that focusing on the influence of training itself, as some authors have done, moves discussion away from the meanings of training for each candidate and from how these meanings bear on the interplay of transference and countertransference in the control analysis. Detailed case examples illustrate how one candidate's experience of training was drawn into his control analyses in the form of enactments, supporting the conclusion that attention to the particular ways in which training influences each candidate's analytic work is critical to the candidate's psychoanalytic education. PMID- 12731805 TI - Change in psychoanalysis: getting from A to B. AB - The change that occurred during an analytic hour with a seven-year-old girl is explored by means of videotape observation and aspects of developmental theory drawn from the field of infant research. Detailed review of the videotape and transcript of twelve minutes of the analytic hour revealed significant change in the girl's attitude toward the loss represented by a cancelled session. The moment-to-moment process between two points, A and B--identified as representing the beginning and end of the change in the girl's ability to deal with loss in the analytic relationship--is studied and described in two forms: written transcript for the verbal material, and narrative description for the nonverbal material. The methodology is based on observed behaviors but also relies on the clinical experience of the analyst in the interpretation of affects and the recognition of intentions in both participants. The results suggest that, taken as a whole, many such small transitional episodes during the course of a treatment play an important role in the process of therapeutic change. PMID- 12731804 TI - Converted patients and clinic patients as control cases: a comparison with implications for psychoanalytic training. AB - All twenty-eight respondents to a recent poll of the thirty institutes affiliated with the American Psychoanalytic Association reported that they now accept cases converted from psychotherapy to psychoanalysis as control cases. This study was designed to compare converted cases to clinic cases systematically with respect to patient characteristics, treatment, and the educational experience of the treating candidate. The study followed twenty-four candidates entering analytic training between 1992 and 1995, who treated thirty-four clinic cases and forty three converted cases between February 1993 and July 2000. Clinic and converted patients were comparable with regard to demographics, prior treatment histories, structural diagnoses, and Axis I diagnoses. In addition, the two groups of cases were indistinguishable with respect to the rate at which candidates received credit toward graduation requirements. Candidates treating converted cases earned approximately dollars 7,600 per patient per year, compared to candidates treating clinic cases, who earned nothing. Eighty-four percent of converted patients diagnosed with a mood disorder by the treating candidate were on medication, in contrast to only 20% of clinic patients with the same diagnosis. Similar differences were seen in the case of anxiety disorders. Given the prevalence of affective and anxiety disorders in control cases and the availability of a variety of medications and psychotherapies with documented efficacy in treating these disorders, candidates should be trained to discuss treatment options with patients who present with Axis I disorders. PMID- 12731806 TI - The course of 253 analyses from selection to outcome. AB - Two studies of the clinical work of experienced psychoanalysts are presented. Study I is a retrospective study of all the analytic work--161 cases--of sixteen analysts from 1973 to 1977, including their evaluations of the treatments at outcome. Study II is a prospective study of the ninety-two cases started in analysis by a group of twenty analysts between 1984 and 1989 and followed to termination, including their reports. The history of each treatment as reported and evaluated by the analyst is examined in terms of: nature of the population, the analyst's view of the case when the initial recommendation was made, duration of treatment, therapeutic benefit, rating of analyzability at termination of treatment, nature of the termination, and cases changed to psychotherapy. Some old shibboleths are challenged as perspective on both the limitations and achievements of psychoanalysis emerges from the findings of the study. PMID- 12731807 TI - Relational perspectives in psychoanalysis. Panel report. PMID- 12731809 TI - Communicating risk: wireless and hardwired. PMID- 12731808 TI - Problems in psychoanalytic education. Panel report. PMID- 12731810 TI - Forest fire risk management and public participation in changing socioenvironmental conditions: a case study in a Mediterranean region. AB - Under current conditions of accelerated socioenvironmental change in the Mediterranean forested landscapes, fire is one of the most critical and difficult risks to tackle within the region. This article summarizes the lessons learned from a project based on the participatory integration of qualitative local stakeholders' knowledge with expert GIS fire simulations carried out in the County of El Bages, Catalonia, Spain. First, in this article, a theoretical model -the forest fire circle--is presented in order to explain the reasons for the rise in the damage and frequency of forest fires in this Mediterranean area. Second, it describes the methodology developed and the stages followed during the project. Results show that: (1) the advocacy of old forest reactive management paradigm assumptions and practices based on uncontrolled forest succession can put vast wooded areas of the Mediterranean basin at critical risk; and (2) forest fire management approaches that ignore the crucial role of long-term prevention and local capacity building strategies have failed. In the final section, the content and the specific dimensions of the old reactive paradigm that has characterized forest fire risk management in Catalonia are discussed and contrasted with the possibly emerging preventative paradigm. PMID- 12731811 TI - A social network contagion theory of risk perception. AB - Risk perceptions have, to a great extent, been studied exclusively as individual cognitive mechanisms in which individuals collect, process, and form perceptions as atomized units unconnected to a social system. These individual-level theories do not, however, help explain how perception of risk may vary between communities or within a single community. One alternative approach is based on a network theory of contagion. This approach, emerging largely from organizational and community social network studies, suggests that it is the relational aspects of individuals and the resulting networks and self-organizing systems that influence individual perceptions and build "groups or communities of like-minded" individuals. These social units, it is argued, behave as attitude, knowledge, or behavioral structures. The study reported in this article tests one aspect of this theoretical perspective. The central hypothesis proposes the existence of risk perception networks--relational groupings of individuals who share, and perhaps create, similar risk perceptions. To test this idea, data were collected from individuals involved in a community environmental conflict over a hazardous waste site cleanup. The statistical analysis used a matrix of relational social linkages to compare with a matrix of individual risk perceptions The analysis confirmed the hypothesis suggesting that social linkages in communities may play an important role in focusing risk perceptions. PMID- 12731812 TI - Evaluating citizen advisory boards: the importance of theory and participant based criteria and practical implications. AB - The role of risk communication and public participation in environmental and public policy decision making has significantly increased over the last 15 years and remains an important social policy issue. In spite of this emphasis, government officials and participants in the process continue to struggle with what makes for "good" public participation. This study used two frameworks--one theoretical and one participant-based-to evaluate two U.S Army Restoration Advisory Boards (RABs). The theoretical framework explores the extent to which the RABs facilitate Habermas's idealized conditions of speech as related to fairness. Not surprisingly, we found that the two RABs do not consistently foster the idealized aspects of fairness suggested by Habermas. The participant-based criteria were elicited through interviews with participants from the various stakeholder groups represented on the RAB, direct observation of RAB meetings, and a review of RAB-related documents. We found that participants' value outcomes (the results of participatory processes) and not just the process itself, which is the focus of the theoretical framework. We also found that participants in the various stakeholder groups had different perceptions of the goals of the participatory process, which were closely related to their notions of success. Our results illustrate both the complexity and importance of using multiple frameworks for evaluating participatory efforts and the need for more systematic evaluation. PMID- 12731813 TI - Using risk communication to disclose the outcome of a participatory decision making process: effects on the perceived acceptability of risk-policy decisions. AB - It has been suggested that public participation during decision making about risks can lead to more widely accepted risk policies. This article discusses an experiment to determine if this is true when people are made aware of the fact that a participatory decision-making process has taken place only through information disclosed during a subsequent risk communication effort. The results from this experiment showed that, after receiving information during risk communication that cast risk policies about space exploration as the product of a participatory decision process, participants in the study felt more supportive of the resulting decisions than did participants in a control group. This result coincided with the participants in the study group perceiving the risks associated with the decision to be lower and the benefits higher. Responses from these participants also showed that they were more satisfied with the decision making process than they were with the outcome of the decision itself Therefore, it may be premature to view the objective of participatory decision-making approaches-and the risk communication efforts that discuss them-as a means of making risk policies more widely acceptable to the public at large. Rather, it may be better to view the benefits of these approaches in terms of their ability to help lead to higher quality decisions that are the product of more widely accepted decision processes. PMID- 12731814 TI - Exploring technical and cultural appeals in strategic risk communication: the Fernald radium case. AB - Risk disputes are often characterized by tensions between technical and cultural understandings of risk and by communication practices that reflect those differing perspectives. This study considers how participants in risk debates draw upon and combine aspects of technical and cultural rationality as broad orientations to risk in expressing their views and formulating persuasive appeals during risk debates. Rhetorical theorist Kenneth Burke's (1984) concept of frames of acceptance is used to analyze a case study involving competing priorities for radium stored at the Fernald site, a former Department of Energy nuclear weapons facility. A rhetorical analysis is conducted using the transcript from a 1995 public meeting during which local residents and a nuclear medicine expert discussed priorities of Fernald site cleanup versus providing radium stored on site for promising cancer research. Two tensions are identified that fostered disagreement among discussants: the first a tension between a local or global context for the controversy and the second a tension between competing definitions of public participation for this issue. This study analyzes the rhetorical strategies by which participants in the Fernald radium debate articulated these tensions and argues that technical and cultural rationality (Plough & Krimsky, 1987) acted as sources of rhetorical invention influencing participants' individual frames of acceptance and the ways they defined and interpreted the situation and crafted persuasive appeals. PMID- 12731815 TI - Determinants of trust in industry, government, and citizen's groups in Japan. AB - The causal structure of the determinants of trust in industry, government, and citizen's groups in Japan was investigated on the basis of Peters et al. (1997). A preliminary survey of the adequacy of the hypotheses proposed by Peters et al. in Japan was made. A set of hypothesized determinants of trust in Japan was proposed based on results of the preliminary survey. Questionnaires concerning perceptions of trust in the organizations and the proposed determinants were sent by mail to residents in the area where environmental risk problems had emerged. The data were analyzed by covariance structure analysis to construct models of trust in industry, government, and citizen's groups. As a result, "openness and honesty," "concern and care," "competence," "people's concern with risks," and "consensual values" were found to be factors directly determining trust. Suggested in particular is that "openness" of an organization is not attained merely by information disclosure, but also by bi-directional communication with the people. Moreover, these models include "consensual values," which do not appear in the model proposed by Peters et al. PMID- 12731816 TI - The use of mental models in chemical risk protection: developing a generic workplace methodology. AB - We adopted a comparative approach to evaluate and extend a generic methodology to analyze the different sets of beliefs held about chemical hazards in the workplace. Our study mapped existing knowledge structures about the risks associated with the use of perchloroethylene and rosin-based solder flux in differing workplaces. "Influence diagrams" were used to represent beliefs held by chemical experts; "user models" were developed from data elicited from open-ended interviews with the workplace users of the chemicals. The juxtaposition of expert and user understandings of chemical risks enabled us to identify knowledge gaps and misunderstandings and to reinforce appropriate sets of safety beliefs and behavior relevant to chemical risk communications. By designing safety information to be more relevant to the workplace context of users, we believe that employers and employees may gain improved knowledge about chemical hazards in the workplace, such that better chemical risk management, self-protection, and informed decision making develop over time. PMID- 12731818 TI - The function of credibility in information processing for risk perception. AB - This study examines how credibility affects the way people process information and how they subsequently perceive risk. Three conceptual areas are brought together in this analysis: the psychometric model of risk perception, Eagly and Chaiken's heuristic-systematic information processing model, and Meyer's credibility index. Data come from a study of risk communication in the circumstance of state health department investigations of suspected cancer clusters (five cases, N = 696). Credibility is assessed for three information sources: state health departments, citizen groups, and industries involved in each case. Higher credibility for industry and the state directly predicts lower risk perception, whereas high credibility for citizen groups predicts greater risk perception. A path model shows that perceiving high credibility for industry and state-and perceiving low credibility for citizen groups-promotes heuristic processing, which in turn is a strong predictor of lower risk perception. Alternately, perceiving industry and the state to have low credibility also promotes greater systematic processing, which consistently leads to perception of greater risk. Between a one-fifth and one-third of the effect of credibility on risk perception is shown to be indirectly transmitted through information processing. PMID- 12731817 TI - Fuzzy-trace theory, risk communication, and product labeling in sexually transmitted diseases. AB - Health care professionals are a major source of risk communications, but their estimation of risks may be compromised by systematic biases. We examined fuzzy trace theory's predictions of professionals' biases in risk estimation for sexually transmitted infections (STIs) linked to: knowledge deficits (producing underestimation of STI risk, re-infection, and gender differences), gist-based mental representation of risk categories (producing overestimation of condom effectiveness for psychologically atypical but prevalent infections), retrieval failure for risk knowledge (producing greater risk underestimation when STIs are not specified), and processing interference involving combining risk estimates (producing biases in post-test estimation of infection, regardless of knowledge). One-hundred-seventy-four subjects (experts attending a national workshop, physicians, other health care professionals, and students) estimated the risk of teenagers contracting STIs, re-infection rates for males and females, and condom effectiveness in reducing infection risk. Retrieval was manipulated by asking estimation questions in two formats, a specific format that "unpacked" the STI category (infection types) and a global format that did not provide specific cues. Requesting estimates of infection risk after relevant knowledge was directly provided, isolating processing effects, assessed processing biases. As predicted, all groups of professionals underestimated the risk of STI transmission, re-infection, and gender differences, and overestimated the effectiveness of condoms, relative to published estimates. However, when questions provided better retrieval supports (specified format), estimation bias decreased. All groups of professionals also suffered from predicted processing biases. Although knowledge deficits contribute to estimation biases, the research showed that biases are also linked to fuzzy representations, retrieval failures, and processing errors Hence, interventions that are designed to improve risk perception among professionals must incorporate more than knowledge dissemination. They should also provide support for information representation, effective retrieval, and accurate processing. PMID- 12731819 TI - Studying heuristic-systematic processing of risk communication. AB - Using a model of risk information seeking and processing developed by Griffin, Dunwoody, and Neuwirth (1999), this study looks at predictors of the processing strategies that people apply to health risk information. Specifically, this article focuses on one relationship within the model--the relationship between perceived amount of information needed to deal with a risk and heuristic systematic processing. Perceived amount of information needed refers to the gap between one's understanding of a risk and the level of understanding that one needs in order to make a decision about that risk. Building on the work of Chaiken (cf. 1980), the Griffin et al. model predicts--and finds--that the larger the gap, the more likely one will process information systematically. The study employs a novel measure of information processing in a survey setting by sending actual information to participants and then asking them how they attended to it; the researchers evaluate this strategy. Finally, the researchers discuss how these findings might help agencies and practitioners create more effective risk messages. PMID- 12731820 TI - Perceptions of recreational fishing boat captains: knowledge and effects of fish consumption advisories. AB - The impacts of fish consumption advisories on recreational and subsistence fishing, particularly in fresh waters, have been examined extensively. By contrast, little attention has focused on organized recreational fishing, such as from party and charter boats, and particularly for salt water fish. We interviewed 93 New Jersey boat captains to determine their knowledge about fish consumption advisories, and whether, in their opinion, clients knew of fish consumption advisories, and whether they thought advisories had an effect on recreational fishing and their businesses. Advisories were ranked by captains as a moderate influence on the success of their business, less so than number of fish caught, strength of the economy, overfishing by commercial boats, and management regulations. Only one boat captain had not heard warnings about eating fish, but what captains said they had heard was mixed in its accuracy and completeness. Clients expect captains to know about fish, and about half of boat captains said clients had asked about the safety of eating fish. Captains who felt advisories were affecting their businesses tended to fish for species without high levels of mercury (except for bluefish) or PCBs, the primary contaminants of concern for state advisories and federal advice. However, these captains worked closer to areas (e.g., Raritan Bay complex and New York Harbor) subject to advisories than did other captains, and were more prone to say that management regulations (e.g., fish size, creel limits, seasons) and marketing and advertising by the industry or state were strong influences on the success of their seasons. Comparing captains who thought advisories had some or great effect (60%) versus those reporting "no effect" (40%), there was no difference in the mean percentage of trips targeting high mercury species such as swordfish and shark. Many captains said they would or might post advisories, but 42% of the boat captains said they would not post consumption warnings if the state provided them. The significant portion (at least 15%) of saltwater fishing supported by these businesses suggests that these captains are an important conduit for future risk communication. PMID- 12731821 TI - The European Commission's White Paper "strategy for a future chemicals policy": a review. AB - The European Commission has proposed a radical new policy for the regulation of chemicals in the EU in the form of a White Paper. The current system has separate regulatory provisions for "new" chemicals (introduced to the market since September 18, 1981) and "existing" chemicals (on the market before September 18,1981). The proposed future policy will have a single unified regulatory system for all chemicals, which should result in better regulation of chemicals in the EU single market. It will be better because risk assessments will be targeted at the chemicals of greatest concern. Furthermore, the system will be streamlined, making regulatory decisions faster, and thus reducing the so-called burden of the past (the large number of chemicals that have never been assessed for their risks to human health or the environment). The new system incorporates the precautionary principle, which will be applied where there is an early indication of unacceptable risk or where there is undue delay in the regulatory process. Moreover, the new strategy is intended to promote greater transparency for all stakeholders. PMID- 12731823 TI - The Commission White Paper on a Strategy for a Future EU Chemicals Policy: the view of European companies of American parentage. AB - After years of good service, EU legislation on chemicals is currently subject to a major review. This process, initiated by the Council of Ministers at Chester in April 1998, will soon lead to new legislative proposals. In the meantime, a review of the Commission's White Paper on "Strategy for a Future Chemicals Policy," published in February 2001, clearly shows that the regulatory landscape in this area will be significantly reshaped and that a new burden will be imposed on industry to demonstrate that the production and use of chemicals indeed conform to high standards of protection of human health and the environment. In the view of industry, on both sides of the Atlantic, while the objectives of the proposed reform can be supported, the measures proposed in the White Paper to implement these objectives are not properly balanced and will lead to substantial societal and economic drawbacks, unless significant adjustments are made. The purpose of this article is to present the pitfalls and difficulties of the reform as they are perceived by the EU Committee of the American Chamber of Commerce in Brussels, an organization that regroups about 150 European companies of American parentage, belonging to a broad range of European business sectors, including producers and users of chemicals. In view of the transatlantic and cross-business character of its membership, the EU Committee offers a different perspective on the debate. PMID- 12731822 TI - The true cost of precautionary chemicals regulation. AB - This article explores the possible social costs of introducing an overly precautionary regulatory regime for chemicals It begins by examining research by the UK Medical Research Council Institute for Environment and Health (MRC-IEH), which suggests that the resource implications of the proposals contained in the European Commission White Paper "Strategy for a Future Chemicals Policy" are unrealistic and even unrealizable. The article then focuses on contemporary debates pertaining to endocrine disrupting chemicals (EDCs) and goes on to question whether a "right to know" is always necessarily a good thing, or whether in certain instances it can lead to a society that feels more sorry than safe. Finally, problems relating to the representation and inclusion of public values in decision-making processes are raised prior to concluding with a call for an ambitious orientation toward social change rather than a self-limiting obsession with safety. PMID- 12731824 TI - The Greens perspective on EU chemicals regulation and the White Paper. PMID- 12731825 TI - Swedish chemical regulation: an overview and analysis. AB - This article begins with a review of the regulation of chemicals in Sweden over the past 30 years, focusing particularly on the 1997 Government Environmental Quality Bill, which called for a toxic-free society by the year 2020. The second part of the article analyzes why Sweden has taken this route. The third and final section discusses Sweden's present role in formulating present EU chemical regulation, such as the recent EU Chemical White Paper, and hypothesizes future impacts of Swedish chemical regulations on the EU itself. PMID- 12731826 TI - Testing alternative decision approaches for identifying cleanup priorities at contaminated sites. AB - This exploratory study compares two approaches for involving nonexpert stakeholders in difficult policy choices. Both approaches have as their goal informing members of the public about contaminated sites and involving them in decisions regarding their cleanup. The first approach focuses on technical information and seeks to improve the available knowledge base so that participants can make choices informed by detailed scientific data. This approach is similar in intent to many of the science-based initiatives in public involvement now being undertaken by EPA, DOE, and other federal or state agencies. The second approach, in contrast, focuses on values-oriented information and seeks to improve stakeholders' ability to make difficult choices in light of required tradeoffs across a variety of technical and nontechnical concerns. The results demonstrate that although both approaches help to increase participants' knowledge level, a values-based approach is more successful in terms of helping nonexpert participants to make decisions aboutwhat have historically been viewed as primarily technical problems. PMID- 12731827 TI - Fuels for urban transit buses: a cost-effectiveness analysis. AB - Public transit agencies have begun to adopt alternative propulsion technologies to reduce urban transit bus emissions associated with conventional diesel (CD) engines. Among the most popular alternatives are emission controlled diesel buses (ECD), defined here to be buses with continuously regenerating diesel particle filters burning low-sulfur diesel fuel, and buses burning compressed natural gas (CNG). This study uses a series of simplifying assumptions to arrive at first order estimates for the incremental cost-effectiveness (CE) of ECD and CNG relative to CD. The CE ratio numerator reflects acquisition and operating costs. The denominator reflects health losses (mortality and morbidity) due to primary particulate matter (PM), secondary PM, and ozone exposure, measured as quality adjusted life years (QALYs). We find that CNG provides larger health benefits than does ECD (nine vs six QALYs annually per 1000 buses) but that ECD is more cost-effective than CNG (dollar 270 000 per QALY for ECD vs dollar 1.7 million to dollar 2.4 million for CNG). These estimates are subject to much uncertainty. We identify assumptions that contribute most to this uncertainty and propose potential research directions to refine our estimates. PMID- 12731828 TI - Making decisions about hazardous waste remediation when even considering a remediation technology is controversial. AB - This paper investigates the circumstances under which proposed hazardous waste remediation technologies are socially acceptable, that is, considered seriously as options in a public arena. First, it summarizes a conceptual framework that guides investigation and interpretation of site-specific remediation decision making. Second, it describes an initial application of that framework to the public participation venue of U.S. DOE Site-Specific Advisory Boards. Investigating the attributes of involved parties and of site context highlights technology acceptability as social decision-making that involves technical and technological issues rather than as a process driven by the technology itself. PMID- 12731829 TI - Open dumping site in Asian developing countries: a potential source of polychlorinated dibenz-p-dioxins and polychlorinated dibenzofurans. AB - Open landfill dumping areas for municipal wastes in Asian developing countries have recently received particular attention with regard to environmental pollution problems. Because of the uncontrolled burning of solid wastes, elevated contamination by various toxic chemicals including dioxins and related compounds in these dumping sites has been anticipated. In this study, concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and coplanar polychlorinated biphenyls (PCBs) were determined in soils from dumping sites in the Philippines, Cambodia, India, and Vietnam. Residue concentrations of PCDD/Fs and coplanar PCBs in dumping site soils were apparently greater than those in soils collected in agricultural or urban areas far from dumping sites, suggesting that dumping sites are potential sources of PCDD/Fs and related compounds. Observed PCDD/F concentrations in soils from dumping sites in the Philippines and Cambodia were comparable or higher than those reported for dioxin-contaminated locations in the world (e.g., near the municipal waste incinerators and open landfill dumping sites). Homologue profiles of PCDD/Fs in dumping site soils from the Philippines and, to a lesser extent, from Cambodia and India reflected patterns of samples representing typical emissions, while profiles of agricultural or urban soils were similar to those of typical environmental sinks. This result suggests recent formation of PCDD/Fs in dumping site areas and that open dumping sites are a potential source of dioxins in Asian developing countries. Uncontrolled combustions of solid wastes by waste pickers, generation of methane gas, and low-temperature burning can be major factors for the formation of dioxins in dumping sites. Elevated fluxes of PCDD/Fs to soils in dumping sites were encountered in the Philippines, Cambodia, India, and Vietnam Hanoi, and these levels were higher than those reported for other countries. Considerable loading rates of PCDD/Fs in the dumping sites of these countries were observed, ranging from 20 to 3900 mg/yr (0.12-35 mg TEQ/yr). PCDD/F concentrations in some soil samples from the Philippines, Cambodia, India, and Vietnam-Hanoi exceeded environmental guideline values, suggesting potential health effects on humans and wildlife living near these dumping sites. The estimated intakes of dioxins via soil ingestion and dermal exposure for children were higher than those for adults, suggesting greater risk of dioxin exposure for children in dumping sites. To our knowledge, this is the first comprehensive study on PCDD/Fs contamination in open dumping sites of Asian developing countries. On the basis of the result of this study, we have addressed a new environmental issue that open dumping sites are potential sources of PCDD/Fs and related compounds, and dioxin contamination in dumping sites may become a key environmental problem in developing countries. PMID- 12731830 TI - Polychlorinated dibenzo-p-dioxins and dibenzofurans in the air of Seveso, Italy, 26 years after the explosion. AB - This study reports the current levels of polychlorinated dibenzo-p-dioxins (PCDDs) and furans (PCDFs) in air at Seveso, where an explosion in a 2,4,5, trichlorophenol production reactor occurred 26 years ago. The aims were to assess if residues of the 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) released during the accident and still present in soil could contaminate the above air and to investigate other potential sources in the area. Long-term air collection was carried out in zones A and B in Seveso and in a reference location in Milan, and samples were analyzed for PCDD and PCDF concentrations by gas chromatography-mass spectrometry (GC-MS). Experimental results showed that no important contribution to the air concentrations is due to the soil contamination and that contemporary sources essentially control the atmospheric burden of PCDDs and PCDFs in the Seveso area. The theoretical release of 2,3,7,8-TCDD from the soils of zones A and B of Seveso was calculated using the SoilFug model. In the worst case, the model simulated an enrichment in atmospheric 2,3,7,8-TCDD concentrations of 4 and 22% for zones A and B, respectively. The investigation of the potential emission sources in the area indicated that combustion of wood residues from furniture factories may be an additional local source of PCDDs and PCDFs. PMID- 12731831 TI - Environmental fate of roxarsone in poultry litter. I. Degradation of roxarsone during composting. AB - Roxarsone, 3-nitro-4-hydroxyphenylarsonic acid, is an organoarsenic compound that is used extensively in the feed of broiler poultry to control coccidial intestinal parasites, improve feed efficiency, and promote rapid growth. Nearly all the roxarsone in the feed is excreted unchanged in the manure. Poultry litter composed of the manure and bedding material has a high nutrient content and is used routinely as a fertilizer on cropland and pasture. Investigations were conducted to determine the fate of poultry-litter roxarsone in the environment Experiments indicated that roxarsone was stable in fresh dried litter; the primary arsenic species extracted with water from dried litter was roxarsone. However, when water was added to litter at about 50 wt % and the mixture was allowed to compost at 40 degrees C, the speciation of arsenic shifted from roxarsone to primarily arsenate in about 30 days. Increasing the amount of water increased the rate of degradation. Experiments also suggested that the degradation process most likely was biotic in nature. The rate of degradation was directly proportional to the incubation temperature; heat sterilization eliminated the degradation. Biotic degradation also was supported by results from enterobacteriaceae growth media that were inoculated with litter slurry to enhance the biotic processes and to reduce the concomitant abiotic effects from the complex litter solution. Samples collected from a variety of litter windrows in Arkansas, Oklahoma, and Maryland also showed that roxarsone originally present had been converted to arsenate. PMID- 12731832 TI - Environmental fate of roxarsone in poultry litter. Part II. Mobility of arsenic in soils amended with poultry litter. AB - Poultry litter often contains arsenic as a result of organo-arsenical feed additives. When the poultry litter is applied to agricultural fields, the arsenic is released to the environment and may result in increased arsenic in surface and groundwater and increased uptake by plants. The release of arsenic from poultry litter, litter-amended soils, and soils without litter amendment was examined by extraction with water and strong acids (HCI and HNO3). The extracts were analyzed for As, C, P, Cu, Zn, and Fe. Copper, zinc, and iron are also poultry feed additives. Soils with a known history of litter application and controlled application rate of arsenic-containing poultry litter were obtained from the University of Maryland Agricultural Experiment Station. Soils from fields with long-term application of poultry litter were obtained from a tilled field on the Delmarva Peninsula (MD) and an untilled Oklahoma pasture. Samples from an adjacent forest or nearby pasture that had no history of litter application were used as controls. Depth profiles were sampled for the Oklahoma pasture soils. Analysis of the poultry litter showed that 75% of the arsenic was readily soluble in water. Extraction of soils shows that weakly bound arsenic mobilized by water correlates positively with C, P, Cu, and Zn in amended fields and appears to come primarily from the litter. Strongly bound arsenic correlates positively with Fe in amended fields and suggests sorption or coprecipitation of As and Fe in the soil column. PMID- 12731833 TI - Trend analysis reveals a recent reduction in mirex concentrations in coho (Oncorhynchus kisutch) and chinook (O. tshawytscha) salmon from Lake Ontario. AB - Lake Ontario, bordering both Canada and the United States, is the only Great Lake with persistent, significant levels of mirex in its biota. Some models suggested that it would take hundreds of years before mirex disappeared from the ecosystem. From 1977 to 1996 the mirex concentrations in coho and chinook salmon greater than 2 kg in weight exceeded the 0.1 mg/kg Food and Drug Administration (FDA) action level for mirex. To determine temporal trends in salmonine mirex levels, slopes and elevations of the regression lines of mirex concentration versus fish weight were compared for each of the six sampling years (1977, 1982, 1986, 1992, 1996, and 1999) by ANCOVA with weight as a covariate. Within 24 years of mirex being banned, mirex least-squares mean concentrations in salmon fillets had decreased significantly. ANCOVA revealed that the slope of the 1999 regression line was significantly flatter (P < or = 0.014) than the slopes of all other regression lines except 1996 (P = 0.966). A Tukey test revealed that the elevation of the 1999 regression line was also significantly lower than all other years (P < 0.001). Based on our results, mirex concentrations in the fillets of most salmon under the size of 12 kg are now below the 0.1 mg/kg United States FDA action level for human consumption. Models suggest that mirex reductions in biota are most likely due to the settling of mirex-contaminated organisms to the sediments and the loss of mirex from the lake through the St Lawrence River. A third mechanism is suggested as the cause of the higher rate of reduction observed in the mid to late 1990s--the control and removal of contaminated groundwater at the former Hooker Chemical site on the Niagara River, the major source of mirex in the watershed of Lake Ontario. PMID- 12731834 TI - Colloid-borne americium migration in Gorleben groundwater: significance of iron secondary phase transformation. AB - The mobility of actinides in natural water may be enhanced by colloid-mediated transport. In this context the reversibility of actinide colloid interaction is a key factor. Iron is an element that can generate colloids under conditions found in natural waters. In this paper, the impact of hematite and the low-crystalline precursor 2-line ferrihydrite on colloid-mediated transport of americium(III) is investigated. Am(III)-containing iron colloids are generated from two different approaches, namely contact between the two in aqueous solution or coprecipitation of Am(III) during iron colloid generation. Dissolved organic carbon (DOC), especially humic substances, has a strong influence on the stability of inorganic colloids. In addition, humic substances interfere in the distribution and kinetics of exchange between groundwater and sediments. Four groundwaters from the Gorleben aquifer system are used with DOC concentrations varying between 0.9 and 81.6 mgC/L together with Pleistocene Aeolian quartz sand from this site. Batch and column experiments are conducted under near-natural conditions (Ar + 1% CO2). To study the influence of kinetics, contact times up to one month are studied. The dynamic light-scattering investigations show that the colloidal stability of the 2-line ferrihydrite increases with increasing DOC concentration. The low-crystalline iron colloids have a marginal influence on the Am(III) transport due to reversibility of americium sorption. Contrary to this, the crystalline hematite generated from coprecipitation of Am(III) leads to an increase of unretarded colloid-mediated Am(III) transport up to a factor of almost five. Chemical characterization of these hematite colloids shows that Am(III) is structurally entrapped in the hematite. The distribution of Am(III) and 2-line ferrihydrite between groundwater and sand sediment remained in disequilibrium even after one month. This shows that the kinetics of Am(III) distribution between the different phases (bulk solution/colloidal form/ sediment) is a key issue. PMID- 12731835 TI - Desorption kinetics for field-aged polycyclic aromatic hydrocarbons from sediments. AB - This study considers desorption kinetics for 12 field-aged polycyclic aromatic hydrocarbons (PAHs) desorbing from size- and density-fractionated sediments collected from two locations in the New York/New Jersey Harbor Estuary. Desorption kinetics for PAHs with a log octanol-water partition coefficient greater than 6 were well-described by a one-domain diffusion model that assumes that PAHs are initially uniformly distributed throughout spherical sediment aggregates. PAH hydrophobicity and sediment specific surface area were the parameters most strongly correlated with the magnitude of the observed diffusivity for the one-domain model. For less hydrophobic PAHs, a two-domain desorption model was used also, and the results suggest that a substantial fraction of these field-aged PAHs desorb via a relatively fast macro-mesopore diffusion mechanism. The model-predicted fraction of PAHs in the fast-diffusion regime by compound and sediment was highly correlated with the measured percent PAH desorption in 24 h. The fast-domain diffusivity was 100 times greater than the slow-domain diffusivity, was correlated with both PAH properties and sediment physical and chemical properties, and could be estimated by readily obtainable physical and chemical parameters. In contrast, the slow-domain diffusivity was not significantly correlated with PAH properties. Our results suggest that macro mesopore diffusion may control mass transport of less-hydrophobic PAHs in estuarine sediments. PMID- 12731836 TI - Intra-aggregate mass transport-limited bioavailability of polycyclic aromatic hydrocarbons to Mycobacterium strain PC01. AB - Biodegradation kinetics for three- and four-ring PAHs by Mycobacterium sp. strain PC01 were measured in whole and density-fractionated estuarine sediments and in a system without intra-aggregate mass transport limitations. The biokinetic data in the systems with and without intra-aggregate mass transport limitations were compared with abiotic PAH desorption kinetics. The results indicate that intra aggregate mass transport limitations, and not the intrinsic bacterial PAH utilization capacity, were most important in controlling the rate of biodegradation of sediment-sorbed PAHs. Achievable extent of biodegradation could be predicted by the independently measured traction of desorbable PAHs in the fast-diffusion regime of a two-domain intra-aggregate mass transport model. A closed-form mathematical model was developed to describe sediment-pore water partitioning and rapid aqueous-phase diffusion of PAHs through the macropore and mesopore network of sediment aggregates, followed by first-order biodegradation of desorbed PAHs in the bulk aqueous domain. The model effectively predicted independent biodegradation kinetics of PAHs field-aged in two estuarine sediments. Despite low aqueous solubility of PAHs, macropore and mesopore diffusion may be an important mechanism controlling intra-aggregate mass transport and bioavailability of the most readily and extensively desorbed PAHs in sediments. PMID- 12731837 TI - Modeling complexometric titrations of natural water samples. AB - Complexometric titrations are the primary source of metal speciation data for aquatic systems, yet their interpretation in waters containing humic and fulvic acids remains problematic. In particular, the accuracy of inferred ambient free metal ion concentrations and parameters quantifying metal complexation by natural ligands has been challenged because of the difficulties inherent in calibrating common analytical methods and in modeling the diverse array of ligands present. This work tests and applies a new method of modeling titration data that combines calibration of analytical sensitivity (S) and estimation of concentrations and stability constants for discrete natural ligand classes ([Li]T and Ki) into a single step using nonlinear regression and a new analytical solution to the one metal/two-ligand equilibrium problem. When applied to jointly model data from multiple titrations conducted at different analytical windows, it yields accurate estimates of S, [Li]T, Ki, and [Cu2+] plus Monte Carlo-based estimates of the uncertainty in [Cu2+]. Jointly modeling titration data at low-and high-analytical windows leads to an efficient adaptation of the recently proposed "overload" approach to calibrating ACSV/CLE measurements. Application of the method to published data sets yields model results with greater accuracy and precision than originally obtained. The discrete ligand-class model is also re-parametrized, using humic and fulvic acids, L1 class (K1 = 10(13) M(-1)), and strong ligands (L(S)) with K(S) >> K1 as "natural components". This approach suggests that Cu complexation in NW Mediterranean Sea water can be well represented as 0.8 +/- 0.3/0.2 mg humic equiv/L, 13 +/- 1 nM L1, and 2.5 +/- 0.1 nM L(S) with [CU]T = 3 nM. In coastal seawater from Narragansett Bay, RI, Cu speciation can be modeled as 0.6 +/- 0.1 mg humic equiv/L and 22 +/- 1 nM L1 or approximately 12 nM L1 and approximately 9 nM L(S), with [CU]T = 13 nM. In both waters, the large excess (approximately 10 nM) of high-affinity, Cu-binding ligands over [CU]T results in low equilibrium [Cu2+] of 10(-14.5 +/- 0.2) M and 10(-13.3 +/- 0.4) M, respectively. PMID- 12731838 TI - Formation of PCDDs, PCDFs, and coplanar PCBs from incineration of various woods in the presence of chlorides. AB - Exhaust gases from the combustion of woods (Japanese red pine, Japanese cedar, Siebold's beech, seawater-impregnated Japanese red pine and Japanese cedar, waste woods containing chlordane, and waste woods containing pentachlorophenol) were collected at the outlet of a combustion chamber. A small-scale incinerator with a stationary grate was used. The samples were analyzed for PCDDs, PCDFs, and coplanar PCBs by gas chromatography/ mass spectrometry (GC/MS). When the grate temperature of the combustion chamber was lower than 700 degrees C, the total amount of PCDDs, PCDFs, and coplanar PCBs formed was proportional to the chlorine content of the combustion samples. On the other hand, when the grate temperature of the combustion chamber was higher than 800 degrees C, there was only a slight formation of PCDDs, PCDFs, and coplanar PCBs regardless of the chlorine content of the combustion samples. When the grate temperature was low, nearly 90% of total PCDDs, PCDFs, and coplanar PCBs formed were PCDFs, whereas when the grate temperature was higher, 50-80% of total PCDDs, PCDFs, and coplanar PCBs formed was PCDFs. The total amount of PCDDs, PCDFs, and coplanar PCBs formed in a high temperature condition was approximately 1/50 of that formed in a low-temperature condition. Coplanar PCBs tended to form less than PCDDs or PCDFs did. Mono-ortho PCBs were formed several times more than nonortho-PCBs. PCDDs or PCDFs contributed significantly to the values of TEQ, while coplanar PCBs contributed only slightly. PMID- 12731839 TI - Environmental ice photochemistry: monochlorophenols. AB - Photolysis of 2- and 4-chlorophenol samples in water ice of the initial concentrations 10(-7) to 10(-2) mol L(-1) is reported. Major phototransformations appeared to be based on the coupling reactions due to chlorophenol aggregation at the grain boundaries of the polycrystalline state. The main products, chlorobiphenyldiols, belong to the family of phenolic halogenated compounds (such as hydroxylated polychlorobiphenyls) that are known xenobiotics found in nature. No photosolvolysis products, that is products from intermolecular reactions between organic and water molecules, were observed at temperatures below -10 degrees C. Raising the temperature to -5 degrees C caused a moderate photosolvolytic activity in the case of 4-chlorophenol (formation of hydroquinone), in contrast to 2-chlorophenol which was almost exclusively transformed into pyrocatechol. It is suggested that photosolvolysis above this temperature occurs in a liquid or quasi-liquid layer that covers the ice crystal surfaces. The results support our model in which significant amounts of some persistent, bioaccumulative, and toxic compounds may be generated by photochemistry of primary pollutants in cold ecosystems and in the upper atmosphere, and may be subsequently released to the environment. PMID- 12731840 TI - Role of soil freezing events in interannual patterns of stream chemistry at the Hubbard Brook Experimental Forest, New Hampshire. AB - Soil freezing is a disturbance of the below ground environment, potentially resulting in increased losses of NO3- and surface water acidification. Here, we report the effects of soil freezing on interannual variation in stream chemistry at the Hubbard Brook Experimental Forest, New Hampshire. Data from 1970 to 1997 of soil frost depth, snow cover, precipitation, air temperature, and stream discharge and chemistry were used in a stepwise linear regression model to select the variables that best predicted deviations of annual stream concentrations from 4-year running averages. Variables quantifying soil freezing severity were selected as significant predictors of short-term fluctuations in stream K+, NO3-, Ca2+, and Mg2+ concentrations from 1970 to 1989, explaining 59 and 47% of the short-term variability in K+ and NO3-, respectively. Fine-root mortality and disturbance of root-soil-microbe interactions, with subsequent effects on decomposition and nutrient uptake, likely contributed to the mobilization of K+ and NO3- to streamwater following severe soil freezing events. The relationship between soil freezing and stream chemistry, however, weakened during the period 1990-1997. Because soil freezing has had inconsistent effects on stream chemistry during the period 1970-1997, it is unclear whether future changes in the frequency, duration, and depth of soil freezing events as the result of changes in the snow cover regime under a warmer climate will have significant impacts on the losses of NO3- and nutrient-base cations from temperate northern ecosystems. PMID- 12731841 TI - Photochemical oxidation of As(III) in ferrioxalate solutions. AB - Photochemical reactions involving aqueous Fe(III) complexes are known to generate free radical species such as OH* that are capable of oxidizing numerous inorganic and organic compounds. Recent work has shown that As(III) can be oxidized to As(V) via photochemical reactions in ferric-citrate solutions; however, the mechanisms of As(III) oxidation and the potential importance of photochemical oxidation in natural waters are poorly understood. Consequently, the objectives of this study were to evaluate oxidation rates of As(III) in irradiated ferrioxalate solutions as a function of pH, identify mechanisms of photochemical As(III) oxidation, and evaluate the oxidation of As(III) in a representative natural water containing dissolved organic C (DOC). The oxidation of As(III) was studied in irradiated ferrioxalate solutions as a function of pH (3-7), As(III), Fe(III), and 2-propanol concentration. Rates of As(III) oxidation (0.5-254 microM h(-1)) were first-order in As(III) and Fe(III) concentration and increased with decreasing pH. Experiments conducted at pH 5.0 using 2-propanol as an OH* scavenger in light and dark reactions suggested that OH* is the important free radical responsible for As(III) oxidation. Significant rates of As(III) oxidation (4-6 microM h(-1)) were also observed in a natural water sample containing DOC, indicating that photochemical oxidation of As(III) may contribute to arsenic (As) cycling in natural waters. PMID- 12731842 TI - A model evaluation of the NO titration technique to remove atmospheric oxidants for the determination of atmospheric organic compounds. AB - Chemical artifact is a problem in the sampling of atmospheric organic species for a relatively long sampling period. In this study, we evaluated a technique for the removal of atmospheric oxidants with added NO during gas and aerosol sampling by theoretical approach using a Regional Atmospheric Chemistry Mechanism (RACM) model. The elimination of O3 in the sample air is regulated predominantly by the reaction of NO and O3 in all simulated cases. We found that, without any oxidant scavenger, OH and NO3 concentrations in the sampler can be kept high even when wall loss processes of radicals are taken into account The relatively high concentration of OH is mainly due to the production of HO(x) in the sample air via the decomposition of HO2NO2 and O3-olefin reactions, whereas NO3 is produced by the decomposition of N2O5. Addition of NO with appropriate concentrations was found to effectively reduce both OH and NO3 concentrations in the sampling devices. This study demonstrates that scavenging of OH and NO3 as well as O3 is important for the study of chemical speciation of organic compounds and that NO addition is a useful technique to eliminate these oxidants. PMID- 12731843 TI - Evaluation of incremental reactivity and its uncertainty in Southern California. AB - The incremental reactivity (IR) and relative incremental reactivity (RIR) of carbon monoxide and 30 individual volatile organic compounds (VOC) were estimated for the South Coast Air Basin using two photochemical air quality models: a 3-D, grid-based model and a vertically resolved trajectory model. Both models include an extended version of the SAPRC99 chemical mechanism. For the 3-D modeling, the decoupled direct method (DDM-3D) was used to assess reactivities. The trajectory model was applied to estimate uncertainties in reactivities due to uncertainties in chemical rate parameters, deposition parameters, and emission rates using Monte Carlo analysis with Latin hypercube sampling. For most VOC, RIRs were found to be consistent in rankings with those produced by Carter using a box model. However, 3-D simulations show that coastal regions, upwind of most of the emissions, have comparatively low IR but higher RIR than predicted by box models for C4-C5 alkenes and carbonyls that initiate the production of HOx radicals. Biogenic VOC emissions were found to have a lower RIR than predicted by box model estimates, because emissions of these VOC were mostly downwind of the areas of primary ozone production. Uncertainties in RIR of individual VOC were found to be dominated by uncertainties in the rate parameters of their primary oxidation reactions. The coefficient of variation (COV) of most RIR values ranged from 20% to 30%, whereas the COV of absolute incremental reactivity ranged from about 30% to 40%. In general, uncertainty and variability both decreased when relative rather than absolute reactivity metrics were used. PMID- 12731844 TI - Estimating the effects of 17alpha-ethinylestradiol on populations of the fathead minnow Pimephales promelas: are conventional toxicological endpoints adequate? AB - Environmental benchmarks have recently been proposed for several steroids including the synthetic steroid, 17alpha-ethinylestradiol (EE2). These benchmarks are based on extrapolation from studies involving long-term exposure of various fish species to EE2. One of the critical studies was a complete life-cycle experiment performed with the fathead minnow Pimephales promelas over a 289 day exposure period. The lowest observed effect concentration (LOEC) and the no observed effect concentration (NOEC) for gonad histology were 4 and 1 ng L(-1) respectively. This was because no testicular tissue could be found in any fish exposed to 4 ng L(-1). In the present paper, the survival and reproduction data from that study are reanalyzed to determine the effects of EE2 on the intrinsic rate of population growth (r = In (lambda)), a parameter of demographic importance. We estimate critical threshold concentrations with respect to r and compare these with those previously derived from conventional toxicity test summaries. Further, we assess the influence of individual variability on threshold estimates using a combination of bootstrap and regression approaches, together with a suite of perturbation analyses. These yield ErC100 values (the concentration estimated to reduce intrinsic growth rate to zero) of 3.11 ng L(-1) (linear model) and 3.41 ng L(-1) (quadratic model), comparable with a maximum acceptable toxicant concentration (MATC) of 2 ng L(-1) for feminization of exposed fish calculated by Laenge et al. Our results indicate that reduction in population growth rate with increasing concentration occurred more through EE2 acting to reduce fertility than survival rates. The significance of these summary statistics when deriving environmental benchmarks for steroid estrogens is discussed in the context of affording protection to populations following long term exposure. PMID- 12731845 TI - Model intercomparison for the uptake of organic chemicals by plants. AB - Currently, a variety of models are available for predicting the uptake, translocation, and elimination of organic contaminants by plants. These models range from simple deterministic risk assessment screening tools to more complex models that consider physical, chemical, and biological processes in a mechanistic manner. This study evaluates the performance of a range of such models and model types against experimental data sets. Three dynamic, three regression-based, and three steady-state and equilibrium models have been selected for evaluation. These models differ in terms of their scope, methodological approach, and complexity. Data from nine published experiments were used to create scenarios to test model performance. These experiments consider plant contamination via both soil and aerial exposure pathways. A total of 19 different organic chemicals were used in the experiments along with 7 different plant species. Model predictions of chemical concentrations in the relevant plant compartments were compared with the experimentally recorded values. The results indicate that dynamic models offer performance advantages for acute exposure durations and for rapidly changing environmental media. Equilibrium/steady-state and regression-based models perform better for chronic exposure durations, where stable conditions are more likely to exist. The selection of an appropriate plant uptake model will therefore be dependent on the requirements of the assessment, the nature of the environmental media, and the duration of the source term. The results generated by the regression-based models suggest that in their current form these models are unsuitable for evaluating the uptake of organic chemicals from the air into plants. PMID- 12731846 TI - Nondestructive, minimal-disturbance, direct-burial solid-phase microextraction fiber technique for measuring TNT in sediment. AB - We explored a novel technique to deploy solid-phase microextraction (SPME) fibers to nondestructively measure the explosive compound 2,4,6-trinitrotoluene (TNT) and its nitroaromatic (NA) degradation products in laboratory sediment toxicity tests and field sediments in situ. SPME fibers within steel mesh envelopes were exposed statically via direct burial within sediment. Six fiber types (polymer coatings) were tested. Polyacrylate (PA) SPME fiber was sufficiently durable for this application, yielded the lowest detection limits, and exhibited a linear uptake relationship across toxicologically relevant sediment NA concentrations (100-2000 nmol/g dw (20-500 microg/g dw)). Temperature greatly influenced SPME absorption kinetics. Via evaluation of absorption at different temperatures, recommended sampling times needed to achieve steady-state equilibrium were 48 h for room temperatures (23-25 degrees C) and up to 7 d for cold (5 degrees C) temperatures. Although a comparison of TNT residues by SPMEs and TNT bioavailability and toxicity in sediments has not been completed, differences in SPME availability of TNT and its degradation products were found between two different TNT-spiked sediments. Our disposable SPME technique was slightly less expensive and as precise as the conventional extraction for total NAs and may prove to be a powerful exposure evaluation tool for assessing the ecological risk of these compounds. PMID- 12731847 TI - Reevaluation of the employment of Fick's law for diffusion dosimeters. AB - This paper reconsiders the means of applying Fick's first law to passive diffusion dosimeters. The performance of the organic vapor monitor (OVM), a commercially available dosimeter, is modeled in terms of gradients, which are generated by evaporating a compound from the dosimeter. The fluxes induced by the gradients are determined gravimetrically. The ratio of a flux and a gradient is reported as a Fick's law proportionality constant, the sampling rate. The sampling rate for the gradient across the OVM is calculated from a harmonic average of the sampling rates of two other gradients. The OVM sampling rates for nine compounds determined by the new methodology agree well with published values. Further analysis of the other two gradients provides a value for an apparent reduction in sampling rate in the absence of airflow across the dosimeter (a boundary-layer effect). Procedures are also described to validate measured air concentrations by determining the sampling rates before and after exposure and by correcting for the boundary-layer effect. Sampling rates were found to be stable during 2-4-day exposures in a variety of conditions. In contrast, the boundary-layer effect caused the measured air concentrations to be substantially lower than the estimated true air concentrations. PMID- 12731848 TI - Residual alcohol influence on NAPL saturation estimates based on partitioning tracers. AB - The influence of residual cosolvent on the partitioning tracer technique for estimating a nonaqueous phase liquid (NAPL) saturation in porous media was investigated. Batch equilibrium and column miscible displacement tests were used to evaluate the influence of residual alcohol cosolvents in the aqueous phase on partitioning and transport of alcohol tracers through sandy soil columns containing tetrachloroethylene (PCE). As the volume fraction of cosolvent alcohol (f(c)) increased ( f(c) < or = 0.1; 10 vol %), partition coefficients (K(nc)) for the alcohol tracers linearly decreased for residual cosolvent ethanol, linearly increased for residual cosolvent tert-butyl alcohol, and did not exhibit an evident change for residual cosolvent 2-propanol. These observations are consistent with measured changes in solubility (S(c)) of the alcohol tracers over the same range (f(c) < or = 0.1) of these residual cosolvent alcohols. Column miscible displacement tests using ethanol as a residual cosolvent ( f(c) < or = 0.1) exhibited earlier partitioning tracer breakthrough leading to an underestimation of NAPL saturation (S(n)) when constant, cosolvent-free partitioning coefficients were assumed. The underestimation magnitude increased with higher initial residual cosolvent alcohol in the columns. The S(n) underestimates were not significant but were 1-10% lower than the actual S(n) (0.18). The estimated partition coefficients based on column tests with residual cosolvent (K(col)) were consistently less than those based on batch tests. Column tests with low (0.5%) and high (15%) S(n) revealed that the residual cosolvent alcohol effect was different depending on the amount of NAPL in the column. Using ethanol for a cosolvent (10%) and 2,4-dimethyl-3-pentanol as a partitioning tracer, the S(n) values were underestimated by about 17% and 5%, respectively, in the low and high NAPL saturation columns. PMID- 12731849 TI - A dynamic numerical model to characterize labile metal complexes collected with diffusion gradient in thin films devices. AB - The speciation of metal species extracted by diffusion gradient in thin films (DGT) devices during experimental deployments in simple metal-ligand synthetic solutions was numerically modeled, analyzed at steady state, and simulated in the dynamic regime. The modeled speciation of two well-known complexes (Cu-citrate and Cu-EDTA) are in good agreement with experimental data obtained in NaNO3 solutions. For any metal complex, the rate at which the metal accumulates on the chelating resin is proportional to the concentration of free metal in solution plus a fraction of the metal complex concentration in solution equivalent to xiD(ML)/D(M). D(M) and D(ML) are the diffusion coefficients of the free and metal complex, respectively, and xi characterizes the complex lability: it is defined as the fraction of metal complex lost when diffusing from the bulk solution to the chelating resin. Numerical simulations were used to explore the variations of xi as a function of several operational and chemical parameters. Lability increases when the dissociation rate constant or the residence time of the metal complex within the hydrogel increase (i.e., by either increasing the thickness of the hydrogel or decreasing D(ML)). Overall, the fraction of metal complex extracted by the DGT decreases when D(ML) decreases, which confirms the ability of diffusion-restrictive hydrogels to separate the free metal fraction from complex solutions. Although the DGT model does not exactly comply with the voltammetric formalism, the lability criteria deltatau(1/2) used with voltammetric macroelectrodes is similar to the lability criteria xi, except that it is valid for high metal-ligand ratio. To determine free metal concentrations in aquatic systems using DGT, it is necessary to account for the presence of numerous ligands with different complexing properties. Soon, the numerical model will be improved to include additional ligands competing for the metal, and new experiments will be designed to discriminate between different labile complexes. Experimental results obtained with different DGT devices (of different hydrogel thicknesses, for example) could be interpreted in terms of conditional kinetic characteristics of the dominant metal-ligand complex in solution. PMID- 12731850 TI - Resolving the unresolved complex mixture in petroleum-contaminated sediments. AB - Comprehensive two-dimensional gas chromatography (GC x GC) was used to investigate the chemical composition of the unresolved complex mixture (UCM) of hydrocarbons in petroleum-contaminated marine sediments. The UCM hydrocarbons were extracted and separated with silica and silver-impregnated silica gel chromatography to yield four fractions (branched alkanes and cycloalkanes, monoaromatics, naphthalenes, and multi-ring PAHs) prior to GC x GC analysis. GC x GC separations used a poly-(dimethylsiloxane) stationary phase for volatility selectivity on the first dimension and a 14% cyanopropylphenyl polysiloxane phase for polarity selectivity on the second dimension to fully resolve monoaromatic, naphthalene, and multi-ring PAH compounds from the UCM. A chiral gamma cyclodextrin phase was used for shape selectivity on the second GC x GC dimension to resolve individual branched alkanes and cycloalkanes in the saturates fraction of the UCM. The ability of GC x GC to resolve thousands of individual chemical components from the UCM will facilitate an understanding of the sources, weathering, and toxicity of UCM hydrocarbons. PMID- 12731851 TI - Fouling and natural organic matter removal in adsorbent/membrane systems for drinking water treatment. AB - Adsorbent particles added to ultrafiltration (UF) systems treating drinking water can remove natural organic matter (NOM) and some other contaminants from the water, but their effect on membrane fouling is inconsistent-in some cases, fouling is reduced, and in others, it is exacerbated. This research investigated the behavior of UF systems to which powdered activated carbon (PAC), heated iron oxide particles (HIOPs), or (nonadsorbent) SiO2 particles were added. On a mass basis, the PAC removed the most NOM from solution, the HIOPs removed less, and the SiO2 removed essentially none. However, in the case of both PAC and SiO2, increasing the dose of solids led to a steady increase in fouling, whereas the opposite trend applied when HIOPs were added. In the absence of NOM, none of the solids fouled the membrane significantly. Thus, even though NOM is a causative agent for fouling, removing it from solution does not necessarily reduce fouling; the mechanism of removal can be just as important as the absolute amount removed, if the removal occurs in a cake layer near the membrane surface. Scanning electron microscopy images of the cake layers formed in the three systems suggest that the NOM binds PAC or SiO2 particles to one another and to the membrane surface, so that the particles become part of the foulant in the system. By contrast, the NOM appears to bind HIOPs to one another but not to the membrane. This process leaves enough pore space in the cake layer for water to reach the membrane with minimal resistance, and it reduces the tendency for either the NOM or the HIOPs to foul the membrane surface. PMID- 12731852 TI - Effect of steric hindrance on carbon dioxide absorption into new amine solutions: thermodynamic and spectroscopic verification through solubility and NMR analysis. AB - Acid gas absorption technology is of great importance in these days for the prevention of global warming and the resulting worldwide climate change. More efficient process design and development for the removal of acid gases has become important, together with the development of new absorbents as one of urgent areas of research in addressing global-warming problems. In the present work, aqueous solutions of 2-amino-2-hydroxymethyl-1,3-propanediol (AHPD), a sterically hindered amine, has been examined as a potential CO2 absorbent and compared with the most commonly used absorbent, monoethanolamine (MEA) solution, through equilibrium solubility measurements and 13C NMR spectroscopic analyses. The solubilities of CO2 in aqueous 10 mass % AHPD solutions were higher than those in aqueous 10 mass % MEA solutions above 4 kPa at 298.15 K, but below 4 kPa, the solubility behavior appeared to be the opposite. The solubility difference between these two solutions increased with the CO2 partial pressures above the crossover pressure. Equilibrated CO2-MEA-H2O and CO2-AHPD-H2O solutions at various CO2 partal pressures ranging from 0.01 to 3000 kPa were analyzed by 13C NMR spectroscopy to provide a more microscopic understanding of the reaction mechanisms in the two solutions. In the CO2-amine-H2O solutions, amine reacted with CO2 to form mainly the protonated amine (AMH+), bicarbonate ion (HCO3-), and carbamate anion (AMCO2-), where the quantitative ratio of bicarbonate ion to carbamate anion strongly influenced the CO2 loading in the amine solutions. A profusion of bicarbonate ions, but a very small amount of carbamate anions, was identified in the CO2-AHPD-H2O solution, whereas a considerable amount of carbamate anions was formed in the CO2-MEA-H2O solution. AHPD contains more hydroxyl groups than nonhindered MEA, and hence, the chemical shifts in its 13C NMR spectra were strongly influenced by the solution pH values. In contrast, MEA appeared to be insensitive to pH. The strong interrelations among CO2 solubility, CO2 partial pressure, bulkiness of the amine structure, and pH identified through the present experimental investigations can provide basic guidelines for finding new potential organic absorbents, including specifically designed amine chemicals. PMID- 12731853 TI - Modes of natural organic matter fouling during ultrafiltration. AB - The fouling of ultrafiltration membranes by natural organic matter (NOM), isolated from a potable surface water source, was studied with an emphasis on elucidating fouling modes and the role of aggregates. NOM size was related to membrane pore sizes using parallel membrane fractionation and size exclusion chromatography, such analyses confirmed the predominance of low MW species and identified the presence of aggregates in concentrated NOM solutions. Cake formation was the dominant mode of fouling by the unfiltered feed, which contained aggregates. This was identified by a constant rate of increase in membrane resistance with permeate throughput and was independent of pore size over a 10-1000 kDa molecular weight cutoff (MWCO) range. Prefiltration (to remove aggregates) and dilution (to reduce aggregate concentration) reduced the rate of increase in membrane resistance for the low MWCO membranes but did not change the fouling mode. In contrast, such pretreatment prevented cake formation on the larger MWCO membranes and shifted the mode of fouling to pore blockage. The date lend support for the idea that an initial fouling layer of large aggregates can catalyze the fouling by lower MW species. The fouling layer could be removed from the large MWCO membranes by backwashing, but the lower MWCO membranes exhibited some irreversible fouling, suggesting that low MW species penetrated into the pore structure. A combined pore blockage-cake formation model described the data well and provided insight into how fouling modes evolve during filtration. PMID- 12731854 TI - Formation of chlorinated aromatics by reactions of Cl*, Cl2, and HCl with benzene in the cool-down zone of a combustor. AB - Conversion of benzene to chlorobenzenes and monochlorophenols by reaction with chlorine radicals (Cl*) in the cool-down zone of a plug-flow combustor has been studied, and a mechanistic analysis of the initial steps of the oxy-chlorination process is proposed. Superequilibrium concentrations of Cl* are formed during combustion of chlorocarbon species and persist at significant concentration levels even after a substantial reduction in the flue gas temperature (T = 500 700 degrees C). At these temperatures, Cl* attack on benzene present in trace concentrations (initial benzene concentration of 300 ppmv or 1080 ppmv were used for the experiments) in the post-flame gas is shown to result in stable chlorinated products (chlorobenzenes and chlorophenols) and loss of benzene. These results suggest that Cl* attack on trace level aromatics and possibly other organic species may be the initial step in the formation of a broad class of chlorinated and oxy-chlorinated pollutants in the post combustion zone. PMID- 12731855 TI - Performance of a constructed wetland with a sulfur/limestone denitrification section for wastewater nitrogen removal. AB - The effectiveness of a nonvegetated lab-scale subsurface flow constructed wetland for wastewater treatment had been evaluated with the feed ammonium concentration of approximately 20-40 mg of NH4(+)-N L(-1) and a hydraulic retention time of approximately 10 d. The present system had a nitrification zone plus a sulfur/limestone (S/L) autotrophic denitrification zone followed by an anaerobic polishing zone and was operated with and without aeration. The wetland had only 80% organics removal and no net nitrogen removal when there was no artificial aeration. However, almost 100% organics removal and approximately 81-90% total inorganic nitrogen (TIN = NH4(+)-N + NO2(-0-N + NO3(-)-N) removal were achieved when the oxic zone of the system was aerated with compressed air. S/L autotrophic denitrification contributed 21-49% of total NO3(-)-N removal across the whole wetland and 50-95% across the S/L column. TIN and NH4(+)-N in the effluent were always < 5.5 and < 0.7 mg L(-1), respectively, when the feed had NH4(+)-N < or = 35 mg L(-1). Sulfate removal of approximately 53-69% was achieved in the anaerobic polishing zone. The position of the S/L column was changed (1.78, 2.24, and 2.69 m from the inlet), and no remarkable difference in nitrogen removal was observed. However, without the S/L column, TIN removal decreased to approximately 74%, and the effluent NO3(-)-N increased about two times (9.13 mg of N L(-1)). The present study has demonstrated the possible use of S/L autotrophic denitrification for nitrate removal in a constructed wetland. PMID- 12731856 TI - Bioreactors for removing methyl bromide following contained fumigations. AB - Use of methyl bromide (MeBr) as a quarantine, commodity, or structural fumigant is under scrutiny because its release to the atmosphere contributes to the depletion of stratospheric ozone. A closed-system bioreactor consisting of 0.5 L of a growing culture of a previously described bacterium, strain IMB-1, removed MeBr (> 110 micromol L(-1)) from recirculating air. Strain IMB-1 grew slowly to high cell densities in the bioreactor using MeBr as its sole carbon and energy source. Bacterial oxidation of MeBr produced CO2 and hydrobromic acid (HBr), which required continuous neutralization with NaOH for the system to operate effectively. Strain IMB-1 was capable of sustained oxidation of large amounts of MeBr (170 mmol in 46 d). In an open-system bioreactor (10-L fermenter), strain IMB-1 oxidized a continuous supply of MeBr (220 /micromol L(-1) in air). Growth was continuous, and 0.5 mol of MeBr was removed from the air supply in 14 d. The specific rate of MeBr oxidation was 7 x 10(-16) mol cell(-1) h(-1). Bioreactors such as these can therefore be used to remove large quantities of contaminant MeBr, which opens the possibility of biodegradation as a practical means for its disposal. PMID- 12731858 TI - Molecular modeling of p-chlorophenoxyacetic acid binding to the CLC-0 channel. AB - Molecular simulation techniques were applied to predict the interaction of the CLC-0 Cl(-) channel and the channel-blocking molecule p-chlorophenoxyacetic acid (CPA). A three-dimensional model of the CLC-0 channel was constructed on the basis of the homology with the bacterial Cl(-) channel StCLC, the structure of which has been solved by X-ray crystallography. Docking of the CPA molecule was obtained by using a geometric recognition algorithm, yielding 5000 possible conformations. By restraining the simulation to those conformations in which CPA is near the intracellular mouth of the channel, the CPA-protein complex models were reduced to three sets of conformations, which are interconvertible within 2 ns when molecular dynamics is applied to the system. Point mutations of CLC-0 at three different positions predicted to interact with CPA in these configurations did, however, not greatly alter CPA inhibition, suggesting a deeper final binding location. In the model, binding of CPA to a more internal position in the ionic pathway was obtained by applying a constant force vector to CPA, pushing it toward the center of the channel. This technique allowed us to outline the possible intrachannel pathway of CPA and to describe qualitatively the binding sites and energy barriers of this pathway. The consistency of the obtained models and the experimental data indicates that the CLC-0-CPA complex model is reasonable and can be used in further biological studies, such as rational design of blocking agents of and mutagenesis of CLC Cl(-) channels. PMID- 12731857 TI - Picosecond dynamics of G-protein coupled receptor activation in rhodopsin from time-resolved UV resonance Raman spectroscopy. AB - The protein response to retinal chromophore isomerization in the visual pigment rhodopsin is studied using picosecond time-resolved UV resonance Raman spectroscopy. High signal-to-noise Raman spectra are obtained using a 1 kHz Ti:Sapphire laser apparatus that provides <3 ps visible (466 nm) pump and UV (233 nm) probe pulses. When there is no time delay between the pump and probe events, tryptophan modes W18, W16, and W3 exhibit decreased Raman scattering intensity. At longer pump-probe time delays of +5 and +20 ps, both tryptophan (W18, W16, W3, and W1) and tyrosine (Y1 + 2xY16a, Y7a, Y8a) peak intensities drop by up to 3%. These intensity changes are attributed to decreased hydrophobicity in the microenvironment near at least one tryptophan and one tyrosine residue that likely arise from weakened interaction with the beta-ionone ring of the chromophore following cis-to-trans isomerization. Examination of the crystal structure suggests that W265 and Y268 are responsible for these signals. These UV Raman spectral changes are nearly identical to those observed for the rhodopsin to-Meta I transition, implying that impulsively driven protein motion by the isomerizing chromophore during the 200 fs primary transition drives key structural changes that lead to protein activation. PMID- 12731859 TI - Protein inhibitors of serine proteinases: role of backbone structure and dynamics in controlling the hydrolysis constant. AB - Standard mechanism protein inhibitors of serine proteinases bind as substrates and are cleaved by cognate proteinases at their reactive sites. The hydrolysis constant for this cleavage reaction at the P(1)-P(1)' peptide bond (K(hyd)) is determined by the relative concentrations at equilibrium of the "intact" (uncleaved, I) and "modified" (reactive site cleaved, I*) forms of the inhibitor. The pH dependence of K(hyd) can be explained in terms of a pH-independent term, K(hyd) degrees, plus the proton dissociation constants of the newly formed amino and carboxylate groups at the cleavage site. Two protein inhibitors that differ from one another by a single residue substitution have been found to have K(hyd) degrees values that differ by a factor of 5 [Ardelt, W., and Laskowski, M., Jr. (1991) J. Mol. Biol. 220, 1041-1052]: turkey ovomucoid third domain (OMTKY3) has K(hyd) degrees = 1.0, and Indian peafowl ovomucoid third domain (OMIPF3), which differs from OMTKY3 by the substitution P(2)'-Tyr(20)His, has K(hyd) degrees = 5.15. What mechanism is responsible for this small difference? Is it structural (enthalpic) or dynamic (entropic)? Does the mutation affect the free energy of the I state, the I* state, or both? We have addressed these questions through NMR investigations of the I and I forms of OMTKY3 and OMIPF3. Information about structure was derived from measurements of NMR chemical shift changes and trans hydrogen-bond J-couplings; information about dynamics was obtained through measurements of (15)N relaxation rates and (1)H-(15)N heteronuclear NOEs with model-free analysis of the results. Although the I forms of each variant are more dynamic than the corresponding I forms, the study revealed no appreciable difference in the backbone dynamics of either intact inhibitor (OMIPF3 vs OMTKY3) or modified inhibitor (OMIPF3* vs OMTKY3*). Instead, changes in chemical shifts and trans-hydrogen-bond J-couplings suggested that the K(hyd) degrees difference arises from differential intramolecular interactions within the intact inhibitors (OMIPF3 vs OMTKY3) in a region of each protein that becomes disordered upon reactive site cleavage (to OMIPF3* and OMTKY3*). PMID- 12731861 TI - Probing the function, conformational plasticity, and dimer-dimer contacts of the GluR2 ligand-binding core: studies of 5-substituted willardiines and GluR2 S1S2 in the crystal. AB - Numerous naturally occurring and synthetic alpha-amino acids act as agonists on (S)-2-amino-3-(3-hydroxy-5-methyl-4-isoxazole) propionic acid (AMPA) receptors but nevertheless display significant differences in their functional properties and modes of interaction. The 5-substituted willardiines are a series of compounds that exhibit a range of affinities, act as partial agonists, and give rise to intermediate levels of activation and desensitization. However, the molecular basis for the activities of 5-substituted willardiines has not been conclusively elaborated at the level of atomic resolution. Here we provide insight into the molecular basis of the potency and efficacy elicited by the 5 substituted willardiines on the basis of cocrystal structures with the GluR2 ligand-binding core. We also show that the crystallized ligand-binding core has an affinity for agonists similar to the ligand-binding core in solution. Analysis of multiple crystal lattices suggests modes by which the ligand-binding core dimers interact in the tetrameric receptor. These studies further our understanding of how subtle differences in the structures of agonists are correlated to changes in the conformation of residues and water molecules in the immediate binding pocket and to the degree of domain closure. PMID- 12731862 TI - Functional and structural consequences of cysteine substitutions in the NH2 proximal region of the human multidrug resistance protein 1 (MRP1/ABCC1). AB - The 190 kDa multidrug resistance protein 1 (MRP1; ABCC1) is comprised of three membrane spanning domains (MSDs) and two nucleotide binding domains (NBDs) configured MSD1-MSD2-NBD1-MSD3-NBD2. MRP1 overexpression in tumor cells results in an ATP-dependent efflux of many oncolytic agents and arsenic and antimony oxyanions. MRP1 also transports GSSG and GSH as well as conjugated organic anions, including leukotriene C(4) and 17beta-estradiol 17-(beta-D-glucuronide) and certain xenobiotics in association with GSH. Previous studies have shown that portions of MSD1 and the cytoplasmic loop (CL3) connecting it to MSD2 are important for MRP1 transport function. In the present study, Cys residues at positions 43, 49, 85, 148, and 190 in MSD1 and positions 208 and 265 in CL3 were mutated to Ala and Ser, and the effects on protein expression, plasma membrane localization, trypsin sensitivity, organic anion transport, and drug resistance properties were investigated. Confocal microscopy showed that 11 of 14 mutants displayed significant levels of nonplasma membrane-associated MRP1. Most mutant proteins were also more resistant to trypsin proteolysis than wild-type MRP1. All Cys mutants transported organic anions (0.5-1.5-fold wild-type MRP1 activity), and cells expressing Ser-substituted but not Ala-substituted Cys43 and Cys265 MRP1 mutants exhibited a 2.5-fold decrease and a 3-fold increase in arsenite resistance, respectively; Cys43Ser MRP1 also conferred lower levels of vincristine resistance. These results indicate that certain Cys residues in the NH(2) proximal region of MRP1 can be important for its structure and selected transport activities. PMID- 12731860 TI - Molecular dynamics, free energy, and SPR analyses of the interactions between the SH2 domain of Grb2 and ErbB phosphotyrosyl peptides. AB - We studied the interactions between the SH2 domain of growth factor receptor binding protein 2 (Grb2) and ErbB receptor-derived phosphotyrosyl peptides using molecular dynamics, free energy calculations, and surface plasmon resonance (SPR) analysis. Binding free energies for nine phosphotyrosyl peptides were calculated using the MM-PBSA continuum solvent method, and excellent qualitative agreement with the SPR experimental data, with a correlation coefficient of 0.92, was obtained. Consistent with previous experimental findings, phosphotyrosyl peptides with the consensus sequence pYXNX showed favorable binding affinity for the Grb2. Unexpectedly, phosphotyrosyl peptides with the consensus sequence pYQQD, which had not shown any specific binding affinity for the Grb2 in earlier studies, also showed favorable binding affinity for the Grb2 in our experimental and computational analyses. Component analysis of the calculated binding free energies revealed that van der Waals interaction between the Grb2 and the phosphotyrosyl peptide was the dominant factor for specificity and binding affinity. These results indicate that current methods of estimating binding free energies are efficient for obtaining important information about protein-protein interactions, which are essential for the transmission of signals in cellular signaling pathways. PMID- 12731863 TI - The structure and specificity of Escherichia coli maltose acetyltransferase give new insight into the LacA family of acyltransferases. AB - The crystallographic three-dimensional structure of the Escherichia coli maa gene product, previously identified as a maltose O-acetyltransferase (MAT) [Brand, B., and Boos, W. (1991) J. Biol. Chem. 266, 14113-14118] has been determined to 2.15 A resolution by the single anomalous dispersion method using data from a crystal cocrystallized with trimethyllead acetate. It is shown here that MAT acetylates glucose exclusively at the C6 position and maltose at the C6 position of the nonreducing end glucosyl moiety. Furthermore, MAT shows higher affinity toward artificial substrates containing an alkyl or hydrophobic chain as well as a glucosyl unit. The presence of a long hydrophobic patch near the acceptor site provides the structural explanation for this preference. The three-dimensional structure reveals the expected trimeric left-handed parallel beta-helix structure found in all other known hexapeptide repeat enzymes. In particular, the structure shows similarities both overall and at the putative active site to the recently determined structure of galactoside acetyltransferase (GAT), the lacA gene product [Wang, X.-G., Olsen, L. R., and Roderick, S. L. (2002) Structure 10, 581 588]. The structure, together with the new biochemical data, suggests that GAT and MAT are more closely related than previously thought and might have similar cellular functions. However, while GAT is specific for acetylation of galactosyl units, MAT is specific for glucosyl units and is able to acetylate maltooligosaccharides, an important property for biotechnological applications. Structural differences at the acceptor site reflect the differences in substrate specificity. PMID- 12731865 TI - Analysis of the leukemia inhibitory factor receptor functional domains by chimeric receptors and cytokines. AB - In contrast to other hematopoietic cytokine receptors, the leukemia inhibitory factor receptor (LIFR) possesses two cytokine binding modules (CBMs). Previous studies suggested that the NH(2)-terminal CBM and the Ig-like domain of the LIFR are most important for LIF binding and activity. Using the recently engineered designer cytokine IC7, which induces an active heterodimer of the LIFR and gp130 after binding to the IL-6R, and several receptor chimeras of the LIFR and the interleukin-6 receptor (IL-6R) carrying the CBM of the IL-6R in place of the COOH terminal LIFR CBM, we could assign individual receptor subdomains to individual binding sites of the ligand. The NH(2)-terminal CBM and the Ig-like domain of the LIFR bind to ligand site III, whereas the COOH-terminal CBM contacts site I. Furthermore, we show that LIFR mutants carrying the IL-6R CBM instead of the COOH terminal CBM can replace the IL-6R by acting as an alpha-receptor for IL-6. However, in situations where a signaling competent receptor is bound at IL-6 site I, ligand binding to site III is an absolute requirement for participation of the receptor in a signaling heterodimer with gp130; i.e., a functional receptor complex of IL-6 type cytokines cannot be assembled solely via site I and II as in the growth hormone receptor complex. PMID- 12731866 TI - Structural hierarchy in molecular films of two class II hydrophobins. AB - Hydrophobins are highly surface-active proteins that are specific to filamentous fungi. They function as coatings on various fungal structures, enable aerial growth of hyphae, and facilitate attachment to surfaces. Little is known about their structures and structure-function relationships. In this work we show highly organized surface layers of hydrophobins, representing the most detailed structural study of hydrophobin films so far. Langmuir-Blodgett films of class II hydrophobins HFBI and HFBII from Trichoderma reesei were prepared and analyzed by atomic force microscopy. The films showed highly ordered two-dimensional crystalline structures. By combining our recent results on small-angle X-ray scattering of hydrophobin solutions, we found that the unit cells in the films have dimensions similar to those of tetrameric aggregates found in solutions. Further analysis leads to a model in which the building blocks of the two dimensional crystals are shape-persistent supramolecules consisting of four hydrophobin molecules. The results also indicate functional and structural differences between HFBI and HFBII that help to explain differences in their properties. The possibility that the highly organized surface assemblies of hydrophobins could allow a route for manufacturing functional surfaces is suggested. PMID- 12731864 TI - Kinetic investigations of the rate-limiting step in human 12- and 15 lipoxygenase. AB - Mammalian lipoxygenases have been implicated in several inflammatory disorders; however, the details of the kinetic mechanism are still not well understood. In this paper, human platelet 12-lipoxygenase (12-hLO) and human reticulocyte 15 lipoxygenase-1 (15-hLO) were tested with arachidonic acid (AA) and linoleic acid (LA), respectively, under a variety of changing experimental conditions, such as temperature, dissolved oxygen concentration, and viscosity. The data that are presented show that 12-hLO and 15-hLO have slower rates of product release (k(cat)) than soybean lipoxygenase-1 (sLO-1), but similar or better rates of substrate capture for the fatty acid (k(cat)/K(M)) or molecular oxygen [k(cat)/K(M(O)2)]. The primary, kinetic isotope effect (KIE) for 15-hLO with LA was determined to be temperature-independent and large ((D)k(cat) = 40 +/- 8), over the range of 10-35 degrees C, indicating that C-H bond cleavage is the sole rate-limiting step and proceeds through a tunneling mechanism. The (D)k(cat)/K(M) for 15-hLO, however, was temperature-dependent, consistent with our previous results [Lewis, E. R., Johansen, E., and Holman, T. R. (1999) J. Am. Chem. Soc. 121, 1395-1396], indicating multiple rate-limiting steps. This was confirmed by a temperature-dependent, k(cat)/K(M) solvent isotope effect (SIE), which indicated a hydrogen bond rearrangement step at low temperatures, similar to that of sLO-1 [Glickman, M. H., and Klinman, J. P. (1995) Biochemistry 34, 14077-14092]. The KIE could not be determined for 12-hLO due to its inability to efficiently catalyze LA, but the k(cat)/K(M) SIE was temperature-independent, indicating distinct rate-limiting steps from both 15-hLO and sLO-1. PMID- 12731867 TI - Stem of SL1 RNA in HIV-1: structure and nucleocapsid protein binding for a 1 x 3 internal loop. AB - The 5'-leader of HIV-1 RNA controls many viral functions. Nucleocapsid (NC) domains of gag-precursor proteins select genomic RNA for packaging by binding several sites in the leader. One is likely to be a stem defect in SL1 that can adopt either a 1 x 3 internal loop, SL1i (including G247, A271, G272, G273) or a 1 x 1 internal loop (G247 x G273) near a two-base bulge (A269-G270). It is likely that these two conformations are both present and exchange readily. A 23mer RNA construct described here models SL1i and cannot slip into the alternate form. It forms a 1:1 complex with NCp7, which interacts most strongly at G247 and G272 (K(d) = 140 nM). This demonstrates that a linear G-X-G sequence is unnecessary for high-affinity binding. The NMR-based structure shows an easily broken G247:A271 base pair. G247 stacks on both of its immediate neighbors and A271 on its 5'-neighbor; G272 and G273 are partially ordered. A bend in the helix axis between the SL1 stems on either side of the internal loop is probable. An important step in maturation of the virus is the transition from an apical loop loop interaction to a dimer involving intermolecular interactions along the full length of SL1. A bend in the stem may be important in relieving strain and ensuring that the strands do not become entangled during the transition. A stem defect with special affinity for NCp7 may accelerate the rate of the dimer transformation. This complex could become an important target for anti-HIV drug development, where a drug could exert its action near a high-energy intermediate on the pathway for maturation of the dimer. PMID- 12731868 TI - Crystal structures of the ribonuclease MC1 mutants N71T and N71S in complex with 5'-GMP: structural basis for alterations in substrate specificity. AB - Ribonuclease MC1 (RNase MC1), isolated from bitter gourd seeds, is a uridine specific RNase belonging to the RNase T2 family. Mutations of Asn71 in RNase MC1 to the amino acids Thr (N71T) and Ser (N71S) in guanosine preferential RNases altered the substrate specificity from uridine specific to guanosine specific, as shown by the transphosphorylation of diribonucleoside monophosphates [Numata, T., et al. (2001) Biochemistry 40, 524-530]. To elucidate the structural basis for the alteration of substrate specificity, crystal structures of the RNase MC1 mutants N71T and N71S, free or complexed with 5'-GMP, were determined at resolutions higher than 2 A. In the N71T-5'-GMP and N71S-5'-GMP complexes, the guanine moiety was, as in the case of the uracil moiety bound to wild-type RNase MC1, firmly stabilized in the B2 site by an extensive network of hydrogen bonds and hydrophobic interactions. Structure comparisons showed that mutations of Asn71 to Thr or Ser cause an enlargement of the B2 site, which then make it feasible to insert a guanine base into the B2 site of mutants N71T and N71S. This binding further allows for hydrogen bonding interaction of the side chain hydroxyl groups of Thr71 or Ser71 with the N7 atom of the guanine base. The mode of guanine binding of mutants N71T and N71S was found to be essentially identical to that of a guanosine preferential RNase NW from Nicotiana glutinosa. In particular, hydrogen bonds between the N7 atom of the guanine base and the hydroxyl groups of the amino acids at position 71 (RNase MC1 numbering) were completely conserved in three guanosine preferential enzymes, thereby indicating that the hydrogen bond may play an essential role in guanine binding in guanosine preferential RNases in the RNase T2 family. Consequently, it can be concluded that amino acids at position 71 (RNase MC1 numbering) serve as one of the determinants for substrate specificity (or preference) in the RNase T2 fimily by changing the size and shape of the B2 site. PMID- 12731869 TI - Temperature dependence of the backbone dynamics of ribonuclease A in the ground state and bound to the inhibitor 5'-phosphothymidine (3'-5')pyrophosphate adenosine 3'-phosphate. AB - The interaction of the dinucleotide inhibitor 5' phosphothymidine(3',5')pyrophosphate adenosine 3'-phosphate (pTppAp) with bovine pancreatic ribonuclease A (RNase A) was characterized by calorimetry and solution NMR spectroscopy. Calorimetric data show that binding of pTppAp to RNase A is exothermic (DeltaH = -60.1 +/- 4.1 kJ/mol) with a dissociation constant of 16 nM at 298 K. At this temperature, the binding results in an entropy loss (TDeltaS = 16.8 +/- 7.3 kJ/mol) that is more favorable than that with the product analogue, 2'-CMP (TDeltaS = -31.3 +/- 0.9 kJ/mol). Temperature-dependent calorimetric experiments give a DeltaC(p) for ligand binding of -230 +/- 100 J/mol K. Binding of pTppAp results in noticeable effects on the backbone amide chemical shifts and dynamics. Amide backbone (15)N NMR spin-relaxation studies were performed on both apo RNase A and RNase A/pTppAp as a function of temperature. At each temperature, the model-free-determined order parameters, S(2), were significantly higher for RNase A/pTppAp than for the apo enzyme indicating a decrease in the conformational entropy of the protein upon ligand binding. Furthermore, the magnitude of this difference varies along the amino acid sequence specifically locating the entropic changes. The temperature dependence of S(2) at each residue enabled assessment of the local heat capacity changes (DeltaC(p)) from ligand binding. In an overall, average sense, DeltaC(p) for the protein backbone, determined from the NMR dynamics measurements, did not differ between apo RNase A and RNase A/pTppAp indicating that backbone dynamics contribute little to DeltaC(p) for protein-ligand interactions in this system. However, residue-by residue comparison of the temperature-dependent change in entropy (DeltaS(B)) between free and bound forms reveals nonzero contributions to DeltaC(p) at individual sites. The balance of positive and negative changes reveals a redistribution of energetics upon binding. Furthermore, experiment and semiempirical estimates suggest that a large negative DeltaC(p) should accompany binding of pTppAp, and we conclude that this contribution must arise from factors other than amide backbone dynamics. PMID- 12731870 TI - Distal site aspartate is essential in the catalase activity of catalase peroxidases. AB - Structural and biochemical characterization of aspartate 152 at the distal heme side of catalase-peroxidase (KatG) from Synechocystis PCC 6803 reveals an important functional role for this residue. In the wild-type protein, the side chain carboxyl group of Asp152 is 7.8 A apart from the heme iron and is hydrogen bonded to two water molecules and a KatG-specific large loop. We have prepared the site-specific variants Asp152Asn, Asp152Ser, Asp152Trp, and Pro151Ala. Exchange of Asp152 exhibited dramatic consequences on the bifunctional activity of this unique peroxidase. The turnover number of catalase activity of Asp152Asn is 2.7%, Asp152Ser 5.7%, and Asp152Trp is 0.6% of wild-type activity. By contrast, the peroxidase activity of the Asp152 variants was 2-7 times higher than that of wild-type KatG or Pro151Ala. The KatG-specific pH profile of the catalase activity was completely different in these variants and exchange of Asp152 made it possible to follow the transition of the ferric enzyme to the redox intermediate compound I by hydrogen peroxide spectroscopically and to determine the corresponding bimolecular rate constant to be 7.5 x 10(6) M(-1) s( 1) (pH 7 and 15 degrees C). The reactivity of compound I toward aromatic one electron donors was enhanced in the Asp152 variants compared with the wild-type protein, whereas the reactivity toward hydrogen peroxide was dramatically decreased. A mechanism for the hydrogen peroxide oxidation, which is different from monofunctional catalases and involves the distal residues Trp122 and Asp152, is proposed. PMID- 12731871 TI - Zinc ion effects on individual Ssp DnaE intein splicing steps: regulating pathway progression. AB - Use of the naturally split, self-splicing Synechocystis sp. PCC6803 DnaE intein permits separate purification of the N- and C-terminal intein domains. Otherwise spontaneous intein-mediated reactions can therefore be controlled in vitro, allowing detailed study of intein kinetics. Incubation of the Ssp DnaE intein with ZnCl(2) inhibited trans splicing, hydrolysis-mediated N-terminal trans cleavage, and C-terminal trans cleavage reactions. Maximum inhibition of the splicing reaction was achieved at equal molar concentrations of ZnCl(2) and intein domains, suggesting a 1:1 metal ion:intein binding stoichiometry. Mutation of the (+)1 cysteine residue to valine (C(+)1V) alleviated the inhibitory effects of ZnCl(2). Valine substitution in the absence of ZnCl(2) blocked trans splicing and decreased C-terminal cleavage kinetics in a manner similar to that of the native (+)1 cysteine in the presence of ZnCl(2). These data are consistent with Zn(2+)-mediated inhibition of the Ssp DnaE intein via chelation of the (+)1 cysteine residue. N-Terminal trans cleavage can occur via both spontaneous hydrolysis and nucleophilic (e.g., DTT) attack. Comparative examination of N terminal cleavage rates using amino acid substitution (C(+)1V) and Zn(2+) mediated inhibition permitted the maximum contribution of hydrolysis to overall N terminal cleavage kinetics to be determined. Stable intermediates consisting of the associated intein domains were detected by PAGE and provided evidence of a rapid C-terminal cleavage step. Acute control of the C-terminal reaction was achieved by the rapid reversal of Zn(2+)-mediated inhibition by EDTA. By inhibiting both the splicing pathway and spontaneous hydrolysis with Zn(2+), reactants can be diverted from the trans splicing to the trans cleavage pathway where DTT and EDTA can regulate N- and C-terminal cleavage, respectively. PMID- 12731872 TI - Kinetic mechanism of the tRNA-modifying enzyme S-adenosylmethionine:tRNA ribosyltransferase-isomerase (QueA). AB - The bacterial enzyme S-adenosylmethionine:tRNA ribosyltransferase-isomerase (QueA) catalyzes the unprecedented transfer and isomerization of the ribosyl moiety of S-adenosylmethionine (AdoMet) to a modified tRNA nucleoside in the biosynthesis of the hypermodified nucleoside queuosine. The complexity of this reaction makes it a compelling problem in fundamental mechanistic enzymology, and as part of our mechanistic studies of the QueA-catalyzed reaction, we report here the elucidation of the steady-state kinetic mechanism. Bi-substrate kinetic analysis gave initial velocity patterns indicating a sequential mechanism, and provided the following kinetic constants: K (M)(tRNA)= 1.9 +/- 0.7 microM and K (M)(AdoMet)= 98 +/- 5.0 microM. Dead-end inhibition studies with the substrate analogues S-adenosylhomocysteine and sinefungin gave competitive inhibition patterns against AdoMet and noncompetitive patterns against preQ(1)-tRNA(Tyr), with K(i) values of 133 +/- 18 and 4.6 +/- 0.5 microM for sinefungin and S adenosylhomocysteine, respectively. Product inhibition by adenine was noncompetitive against both substrates under conditions with a subsaturating cosubstrate concentration and uncompetitive against preQ(1)-tRNA(Tyr) when AdoMet was saturating. Inhibition by the tRNA product (oQ-tRNA(Tyr)) was competitive and noncompetitive against the substrates preQ(1)-tRNA(Tyr) and AdoMet, respectively. Inhibition by methionine was uncompetitive versus preQ(1)-tRNA(Tyr), but noncompetitive against AdoMet. However, when methionine inhibition was investigated at high AdoMet concentrations, the pattern was uncompetitive. Taken together, the data are consistent with a fully ordered sequential bi-ter kinetic mechanism in which preQ(1)-tRNA(Tyr) binds first followed by AdoMet, with product release in the order adenine, methionine, and oQ-tRNA. The chemical mechanism that we previously proposed for the QueA-catalyzed reaction [Daoud Kinzie, S., Thern, B., and Iwata-Reuyl, D. (2000) Org. Lett. 2, 1307-1310] is consistent with the constraints imposed by the kinetic mechanism determined here, and we suggest that the magnitude of the inhibition constants for the dead-end inhibitors may provide insight into the catalytic strategy employed by the enzyme. PMID- 12731873 TI - Allosteric activator domain of maintenance human DNA (cytosine-5) methyltransferase and its role in methylation spreading. AB - The human maintenance DNA (cytosine-5) methyltransferase (hDNMT1) consists of a large N-terminal regulatory domain fused to a catalytic C-terminal domain by randomly repeated Gly-Lys dipeptides. Several N-terminal deletion mutants of hDNMT1 were made, purified, and tested for substrate specificity. Deletion mutants lacking 121, 501, 540, or 580 amino acids from the N-terminus still functioned as DNA methyltransferases, methylated CG sequences, and preferred hemimethylated to unmethylated DNA, as did the full-length hDNMT1. Methylated DNA stimulated methylation spreading on unmethylated CpG sequences for the full length and the 121 amino acid deletion hDNMT1 equally well but not for the mutants lacking 501, 540, or 580 amino acids, indicating the presence of an allosteric activation determinant between amino acids 121 and 501. Peptides from the N- and C-termini bound methylated DNA independently. Point mutation analysis within the allosteric region revealed that amino acids 284-287 (KKHR) were involved in methylated DNA-mediated allosteric activation. Allosteric activation was reduced in the double point mutant enzymes D25 (K284A and K285A) and D12 (H286A and R287A). Retinoblastoma gene product (Rb), a negative regulator of DNA methylation, bound to the allosteric site of hDNMT1 and inhibited methylation, suggesting Rb may regulate methylation spreading. PMID- 12731874 TI - Thermodynamic characterization of the binding of nucleotides to glycyl-tRNA synthetase. AB - The interaction of adenine nucleotides with glycyl-tRNA synthetase was examined by several experimental approaches. ATP and nonsubstrate ATP analogues render glycyl-tRNA synthetase more resistant to digestion by a number of proteases (thrombin, Arg-C, and chymotrypsin) at concentrations that correlate with their Michaelis constants or inhibition constants, consistent with their exerting an effect by binding at the ATP site. Glycine had little effect alone but potentiated the effect of ATP in increasing the resistance to thrombin digestion, consistent with the formation of an enzyme-bound adenylate. No protection from thrombin digestion was afforded by tRNA(gly). Binding constants were determined by isothermal titration calorimetry at 25 degrees C for ATP (2.5 x 10(5) M(-1)), AMPPNP (3.7 x 10(5) M(-1)), and AMPPCP (2.2 x 10(6) M(-1)). The nucleotides had similar values for DeltaH (-71 kJ mol(-1)), with values for TDeltaS that accounted for the differences in the binding constants. Near-ultraviolet CD spectra of the enzyme-nucleotide complexes indicate that the nucleotides are bound in the anti configuration. A glycyl-adenylate analogue, glycine sulfamoyl adenosine (GSAd), bound with a large value for DeltaH (-187 kJ mol(-1)), which was balanced by a large TDeltaS term to give a binding constant (3.7 x 10(6) M( 1)) only slightly larger than that of AMPPCP. Glycine binding to the enzyme could not be detected calorimetrically, and its presence did not change the thermodynamic parameters for binding of AMPPCP. AMPPNP and AMPPCP were not substrates for glycyl-tRNA synthetase. Analysis of the temperature dependence of ATP binding indicated that the heat capacity change is small, whereas the binding of GSAd is accompanied by a large negative heat capacity change (-2.6 kJ K(-1) mol(-1)). Titrations performed in buffers with different ionization enthalpies indicate that the large value for DeltaH for the adenylate analogue does not arise from a coupled protonation event. Differential scanning calorimetry indicated that glycyl-tRNA synthetase is stabilized by nucleotides. Unfolding of the protein is irreversible, and thermodynamic parameters for unfolding could therefore not be determined. The results are consistent with a significant conformational transition in glycyl-tRNA synthetase coupled to the binding of GSAd. PMID- 12731875 TI - Interfacial anchor properties of tryptophan residues in transmembrane peptides can dominate over hydrophobic matching effects in peptide-lipid interactions. AB - Membrane model systems consisting of phosphatidylcholines and hydrophobic alpha helical peptides with tryptophan flanking residues, a characteristic motif for transmembrane protein segments, were used to investigate the contribution of tryptophans to peptide-lipid interactions. Peptides of different lengths and with the flanking tryptophans at different positions in the sequence were incorporated in relatively thick or thin lipid bilayers. The organization of the systems was assessed by NMR methods and by hydrogen/deuterium exchange in combination with mass spectrometry. Previously, it was found that relatively short peptides induce nonlamellar phases and that relatively long analogues order the lipid acyl chains in response to peptide-bilayer mismatch. Here it is shown that these effects do not correlate with the total hydrophobic peptide length, but instead with the length of the stretch between the flanking tryptophan residues. The tryptophan indole ring was consistently found to be positioned near the lipid carbonyl moieties, regardless of the peptide-lipid combination, as indicated by magic angle spinning NMR measurements. These observations suggest that the lipid adaptations are not primarily directed to avoid a peptide-lipid hydrophobic mismatch, but instead to prevent displacement of the tryptophan side chains from the polar-apolar interface. In contrast, long lysine-flanked analogues fully associate with a bilayer without significant lipid adaptations, and hydrogen/deuterium exchange experiments indicate that this is achieved by simply exposing more (hydrophobic) residues to the lipid headgroup region. The results highlight the specific properties that are imposed on transmembrane protein segments by flanking tryptophan residues. PMID- 12731876 TI - Analysis of tubulin isotypes and mutations from taxol-resistant cells by combined isoelectrofocusing and mass spectrometry. AB - Six human alpha-tubulin and seven human beta-tubulin isotypes, each of which can undergo posttranslational modifications, have been detected by the reverse transcriptase-polymerase chain reaction. This repertoire of tubulin isotypes plays a role in development and in the building of specialized microtubule-based structures. In cell lines, the relationship between resistance to microtubule interacting drugs and altered tubulin isotype expression profiles is often established by quantitation of cDNA and/or Western blot analysis. Tubulin mutations in major isotypes are detected by sequencing cDNA, but more analysis of expression of tubulin mutations at the protein level, to assess their role in drug resistance, is needed. We utilized a Taxol-based purification and high resolution isoelectrofocusing combined with a mass spectrometry-based analysis of tubulin. This approach has allowed the separation and relative quantitation of tubulin isotypes having a difference in isoelectric point values of 0.01, without the need for two-dimensional gel electrophoresis. The specificity of tubulin isotype antibodies also has been established. In cell lines resistant to microtubule-stabilizing drugs that express heterozygous tubulin mutations, the relative amount of mutant tubulin expression has been determined. In these cell lines, the absence of betaII- and betaIVa-tubulin has been demonstrated, and an increased level of expression of betaIII-tubulin in resistant cells has been confirmed, indicating that this tubulin isotype is a unique marker of resistance. PMID- 12731877 TI - Insights into the association of FcgammaRII and TCR with detergent-resistant membrane domains: isolation of the domains in detergent-free density gradients facilitates membrane fragment reconstitution. AB - Plasma membrane rafts are routinely isolated as detergent-resistant membranes (DRMs) floating in detergent-free density gradients. Here we show that both the presence and exclusion of TX-100 during the density gradient fractionation have profound effects on the location of FcgammaRII and TCR in DRM fractions. The presence of TX-100 during fractionation promoted solubilization of non-cross linked FcgammaRII when the receptor was insufficiently dissolved upon cell lysis. In the detergent-supplemented gradients, TX-100 micelles floated, further enhancing dissociation of FcgammaRII and TCR from DRMs and promoting a shift of the receptors toward higher-density fractions. Hence, fractionation of cell lysates over the detergent-containing gradients enables isolation of DRMs devoid of weakly associated proteins, like nonactivated FcgammaRII and TCR. On the other hand, in a detergent-free gradient, non-cross-linked FcgammaRII, fully soluble in 0.2% TX-100, was recovered in DRM fractions. Moreover, employment of the TX-100 free gradient for refractionation of intermediate-density fractions, derived from detergent-supplemented gradients and containing FcgammaRII and TCR, resulted in flotation of the receptors to buoyant fractions. An analysis of the TX-100 concentration revealed that after fractionation of 0.2% TX-100 cell lysates in the absence of detergent, the level of TX-100 in DRM fractions was reduced to 0.01%, below the critical micelle concentration. Therefore, fractionation of detergent cell lysates over detergent-free gradients can mimic conditions for a membrane reconstitution, evoking association of a distinct subset of membrane proteins, including FcgammaRII and TCR, with DRMs. PMID- 12731878 TI - Channel formation by the binding component of Clostridium botulinum C2 toxin: glutamate 307 of C2II affects channel properties in vitro and pH-dependent C2I translocation in vivo. AB - The binding component (C2II) of the binary Clostridium botulinum C2 toxin mediates transport of the actin ADP-ribosylating enzyme component (C2I) into the cytosol of target cells. C2II (80 kDa) is activated by trypsin cleavage, and proteolytically activated C2II (60 kDa) oligomerizes to heptamers in solution. Activated C2II forms channels in lipid bilayer membranes which are highly cation selective and voltage-gated. A role for this channel in C2I translocation across the cell membrane into the cytosol is discussed. Amino acid residues 303-331 of C2II contain a conserved pattern of alternating hydrophobic and hydrophilic residues, which likely facilitates membrane insertion and channel formation by creating two antiparallel beta-strands. Some of the residues are in strategic positions within the putative C2II channel, in particular, glutamate 307 (E307) localized in its center and glycine 316 (G316) localized on the trans side of the membrane. Here, single-lysine substitutions of these amino acids and the double mutant E307K/G316K of C2II were analyzed in vivo and in artificial lipid bilayer experiments. The pH dependence of C2I transport across cellular membranes was altered, and a pH of or= -200 mV). The DeltaP34 variant retains high conformational stability and remains a substrate for thioredoxin reductase. In contrast to the reduced form of the wild-type enzyme, the reduced form of DeltaP34 Trx has disulfide isomerization activity, which is 25-fold greater than that of the CGC peptide. Thus, the rational deletion of an active site residue can bestow a new and desirable function upon an enzyme. Moreover, a CXC motif, in both a peptide and a protein, provides functional mimicry of PDI. PMID- 12731882 TI - Transient kinetic studies of heme reduction in Escherichia coli nitrate reductase A (NarGHI) by menaquinol. AB - We have studied the transient kinetics of quinol-dependent heme reduction in Escherichia coli nitrate reductase A (NarGHI) by the menaquinol analogue menadiol using the stopped-flow method. Four kinetic phases are observed in the reduction of the hemes. A transient species, likely to be associated with a semiquinone radical anion, is observed with kinetics that correlates with one of the phases. The decay of the transient species and the formation of the second reduction phase of the hemes can be fitted to a double-exponential equation giving similar rate constants, k(1) = 9.24 +/- 0.9 s(-1) and k(2) = 0.22 +/- 0.02 s(-1) for the decay of the transient species, and k(1) = 9.23 +/- 0.9 s(-1) and k(2) = 0.22 +/- 0.02 s(-1) for the formation of the reduction phase. The quinol-binding-site inhibitors 2-n-heptyl-4-hydroxyquinoline-N-oxide (HOQNO) and stigmatellin have significant and different inhibitory effects on the reduction kinetics. The kinetics of heme reduction in NarI expressed in the absence of the NarGH catalytic dimer (NarI(DeltaGH) exhibits only two kinetic phases, and the decay of the transient species also correlates kinetically with the second reduction phase of the hemes. We have also studied nitrate-dependent heme reoxidation following quinol-dependent heme reduction using a sequential stopped-flow method. HOQNO elicits a much stronger inhibitory effect than stigmatellin on the reoxidation of the hemes. On the basis of our results, we propose schemes for the mechanism of NarGHI reduction by menaquinol and reoxidation by nitrate. PMID- 12731883 TI - Tumor-associated trypsinogen-2 (trypsinogen-2) activates procollagenases (MMP-1, 8, -13) and stromelysin-1 (MMP-3) and degrades type I collagen. AB - A critical step in cancer growth and metastasis is the dissolution of the extracellular matrix surrounding the malignant tumor, which leads to tumor cell invasion and dissemination. Type I collagen degradation involves the initial action of collagenolytic matrix metalloproteinases (MMP-1, -8, and -13) activated by MMP-3 (stromelysin-1). The role of interactive matrix serine proteinases (MSPs), including tumor-associated trypsinogens, has been unclear in collagenolysis. Now, we provide evidence that the major isoenzyme of human tumor associated trypsinogens, trypsin-2, can directly activate three collagenolytic proMMPs as well as proMMP-3. These proMMP activations are inhibited by tumor associated trypsin inhibitor (TATI). Furthermore, we demonstrate that trypsin-2 efficiently degrades native soluble type I collagen, which can be inhibited by TATI. However, cell culture studies showed that trypsin-2 transfection into the HSC-3 cell line did not result in MMP-1, -3, -8, and -13 activation but affected MMP-3 and -8 production at the protein level. These findings indicate that human trypsin-2 can be regarded as a potent tumor-associated matrix serine protease capable of being the initial activator of the collagenolytic MMP activation network as well as directly attacking type I collagen. PMID- 12731884 TI - Interlobe communication in human serum transferrin: metal binding and conformational dynamics investigated by electrospray ionization mass spectrometry. AB - Human serum transferrin (hTF) is an iron transport protein, comprising two lobes (N and C), each containing a single metal-binding center. Despite substantial structural similarity between the two lobes, studies have demonstrated the existence of significant differences in their metal-binding properties. The nature of these differences has been elucidated through the use of electrospray ionization mass spectrometry to study both metal retention and conformational properties of hTF under a variety of conditions. In the absence of chelating agents or nonsynergistic anions, the diferric form of hTF remains intact until the pH is lowered to 4.5. The monoferric form of hTF retains the compact conformation until the pH is lowered to 4.0, whereas the apoprotein becomes partially unfolded at pH as high as 5.5. Selective (lobe-specific) modulation of the iron-binding properties of hTF using recombinant forms of the protein (in which the pH-sensitive elements in each lobe were mutated) verifies that the N lobe of the protein has a lower affinity for ferric ion. Surprisingly, the apo-N lobe is significantly less flexible compared to the apo-C-lobe. Furthermore, the conformation of the iron-free N-lobe is stabilized when the C-lobe contains iron, confirming the existence of an interlobe interaction within the protein. The experimental results provide strong support for the earlier suggestion that hTF interacts with its receptor (TFR) primarily through the C-lobe both at the cell surface and inside the endosome. PMID- 12731885 TI - Multidrug resistance protein (MRP) 1 and MRP3 attenuate cytotoxic and transactivating effects of the cyclopentenone prostaglandin, 15-deoxy Delta(12,14)prostaglandin J2 in MCF7 breast cancer cells. AB - One of the most potent cyclopentenone prostaglandins, 15-deoxy Delta(12,14)prostaglandin J(2) (15-d-PGJ(2)), has been shown to be cytotoxic in some tumor cells and, as a ligand of peroxisome proliferator activated receptor gamma (PPARgamma), to influence the transcriptional regulation of several genes. We examined whether a glutathione conjugate of 15-d-PGJ(2), 15-d-PGJ(2)-SG, is formed and if the glutathione conjugate efflux pumps, MRP1 and MRP3, could transport this conjugate, thereby attenuating the cytotoxicity and transactivating activity of 15-d-PGJ(2) in MCF7 breast cancer cells. Formation of 15-d-PGJ(2)-SG was demonstrated both in vitro and in cells, and its structure was determined by ESI/MS and NMR. Expression of MRP1 and MRP3 was achieved by stable transduction of parental MCF7 cells. Membrane vesicles derived from these cells supported efficient, ATP-dependent transport of 15-d-PGJ(2)-SG (K(M) 1.4 and 2.9 microM for MRP1 and MRP3, respectively). When compared with parental, MRP-minus MCF7 cells, expression of MRP1 and MRP3 conferred approximately 2-fold protection from 15-d-PGJ(2) cytotoxicity. 15-d-PGJ(2)-mediated transcriptional activation was evaluated in cells transiently transfected with a reporter gene under the transcriptional control of a PPAR responsive element. Treatment of parental MCF7 cells with 15-d-PGJ(2) resulted in a time-dependent induction of reporter gene activity-induction that was measurable with concentrations of added 15-d-PGJ(2) as low as 100 nM. In contrast, expression of MRP1 or MRP3 abolished 15-d-PGJ(2) dependent reporter gene induction. Depletion of intracellular glutathione reversed MRP1- and MRP3-mediated attenuation of 15-d-PGJ(2) cytotoxicity and transactivation. These data indicate that MRP1 and MRP3 can modulate the biological effects of 15-d-PGJ(2), and likely other cyclopentenone prostaglandins, in a glutathione-dependent manner. The results are consistent with a mechanism for the attenuation of the biological activities of 15-d-PGJ(2) that involves the formation and active efflux of its glutathione conjugate, 15-d PGJ(2)-SG. PMID- 12731886 TI - Unmasking tandem site interaction in human acetylcholinesterase. Substrate activation with a cationic acetanilide substrate. AB - Acetylcholinesterase (AChE) contains a narrow and deep active site gorge with two sites of ligand binding, an acylation site (or A-site) at the base of the gorge, and a peripheral site (or P-site) near the gorge entrance. The P-site contributes to catalytic efficiency by transiently binding substrates on their way to the acylation site, where a short-lived acyl enzyme intermediate is produced. A conformational interaction between the A- and P-sites has recently been found to modulate ligand affinities. We now demonstrate that this interaction is of functional importance by showing that the acetylation rate constant of a substrate bound to the A-site is increased by a factor a when a second molecule of substrate binds to the P-site. This demonstration became feasible through the introduction of a new acetanilide substrate analogue of acetylcholine, 3 (acetamido)-N,N,N-trimethylanilinium (ATMA), for which a = 4. This substrate has a low acetylation rate constant and equilibrates with the catalytic site, allowing a tractable algebraic solution to the rate equation for substrate hydrolysis. ATMA affinities for the A- and P-sites deduced from the kinetic analysis were confirmed by fluorescence titration with thioflavin T as a reporter ligand. Values of a >1 give rise to a hydrolysis profile called substrate activation, and the AChE site-specific mutant W86F, and to a lesser extent wild type human AChE itself, showed substrate activation with acetylthiocholine as the substrate. Substrate activation was incorporated into a previous catalytic scheme for AChE in which a bound P-site ligand can also block product dissociation from the A-site, and two additional features of the AChE catalytic pathway were revealed. First, the ability of a bound P-site ligand to increase the substrate acetylation rate constant varied with the structure of the ligand: thioflavin T accelerated ATMA acetylation by a factor a(2) of 1.3, while propidium failed to accelerate. Second, catalytic rate constants in the initial intermediate formed during acylation (EAP, where EA is the acyl enzyme and P is the alcohol leaving group cleaved from the ester substrate) may be constrained such that the leaving group P must dissociate before hydrolytic deacylation can occur. PMID- 12731887 TI - A glycosylation site, 60SGTS63, of p67 is required for its ability to regulate the phosphorylation and activity of eukaryotic initiation factor 2alpha. AB - Eukaryotic initiation factor 2- (eIF2-) associated glycoprotein p67 blocks eIF2alpha phosphorylation by kinases, and its N-terminal 1-97 amino acid segment can induce efficient translation. To investigate whether glycosylation at the serine/threonine clusters at this region is important in protein synthesis, we selected (27)TSST(30) and (60)SGTS(63) clusters for further analysis. By site directed mutagenesis, (27)TSST(30) and (60)SGTS(63) clusters were substituted with (27)AAGA(30) and (60)AGAA(63) amino acid residues in full-length p67, and their EGFP fusions were constitutively expressed in rat tumor hepatoma cells (KRC 7). The (60)AGAA(63) mutant blocked eIF2alpha phosphorylation less than either wild-type p67 or the (27)AAGA(30) mutant. The (60)AGAA(63) mutant also showed a low level of protein synthesis rate, a lower level of glycosylation, increased turnover rate, and weaker binding to eIF2alpha. These results suggest that glycosylation within the (60)SGTS(63) sequence of p67 plays an important role in its stability and thus its regulation of protein synthesis by modulating the phosphorylation of the alpha-subunit of eIF2. PMID- 12731888 TI - Chemical dissection of the effects of tyrosine phosphorylation of SHP-2. AB - The regulation of the protein tyrosine phosphatase (PTPase) SHP-2 by tyrosine phosphorylation has been difficult to elucidate because of the intrinsic instability of the phosphoprotein. In the past, expressed protein ligation has been used to site-specifically incorporate the phosphotyrosine mimic Pmp (phosphonomethylene phenylalanine) into the two tyrosine phosphorylation sites (542, 580) of SHP-2 one at a time to analyze the effects on catalytic behavior. In this study, we have incorporated two Pmps into the phosphorylation sites simultaneously and examined the effects of double SHP-2 tyrosine phosphorylation. We have found that the Pmp groups show close to additive effects on PTPase stimulation, suggesting dual SH2 domain occupancy. The relative effects of the phosphotyrosine analogue difluoromethylene phosphonophenylalanine (F(2)Pmp) compared to those of Pmp were also examined. It was found that the F(2)Pmp analogue showed slightly enhanced PTPase stimulation compared with the Pmp analogue, consistent with its higher affinity for SH2 domains. Taken together with the bis-Pmp studies, these data suggest that double phosphorylation of the SHP-2 C-terminus could give rise to a 9-fold overall PTPase activation, 30-50% of the value associated with deletion of the SH2 domains. Catalytically inactive forms of phosphorylated SHP-2 proteins were also produced by expressed protein ligation. This allowed for a systematic analysis of intermolecular autodephosphorylation of SHP-2, which revealed how conformational plasticity can modulate phosphotyrosine stability. PMID- 12731889 TI - Flavin redox state triggers conformational changes in the PutA protein from Escherichia coli. AB - The regulation of proline utilization in Escherichia coli involves the proline dependent translocation of the PutA flavoprotein from the cytoplasm to a peripheral position on the membrane. In the cytoplasm, PutA represses transcription of the proline utilization (put) genes while membrane-bound PutA catalyzes the oxidation of L-proline to glutamate. The mechanism by which PutA switches from a DNA-binding protein to a membrane-bound enzyme involves a proline induced conformational change that is characterized by the appearance of a 119 kDa fragment during limited proteolysis of proline-reduced PutA. To establish whether the FAD redox state is responsible for the proline-induced conformational change in PutA, we distinguished the effects that FAD reduction and proline analogue binding have on PutA conformation by limited chymotrypsin proteolysis. Controlled potentiometric proteolysis of PutA demonstrated that the formation of the 119-kDa band occurs at an E(m)(conf) value of -0.058 V (pH 7.5), which is within 20 mV of the E(m) value for FAD bound to PutA. The manipulation of the E(m)(conf) value by reconstitution of PutA with the FAD analogue, 5-deazaFAD, confirmed that the conformational change observed in the presence of proline is solely dependent on the FAD redox state. The proline analogue, L-tetrahydro-2 furoic acid (L-THFA), failed to elicit the formation of the 119-kDa fragment during chymotrypsin cleavage of PutA. Instead, a unique fragment of about 93-kDa was observed, indicating that a distinct PutA conformer is stabilized by L-THFA. Reduction of L-THFA-complexed PutA, however, regenerated the 119-kDa fragment showing that reduction of the FAD cofactor overrides conformational changes induced by L-THFA. Mapping of the protease susceptibility sites in PutA revealed that the conformational changes caused by FAD reduction and L-THFA binding are transmitted to domains outside the proline dehydrogenase active site. PMID- 12731891 TI - The mitochondrial oxoglutarate carrier: structural and dynamic properties of transmembrane segment IV studied by site-directed spin labeling. AB - The structural and dynamic features of the fourth transmembrane segment of the mitochondrial oxoglutarate carrier were investigated using site-directed spin labeling and electron paramagnetic resonance (EPR). Using a functional carrier protein with native cysteines replaced with serines, the 18 consecutive residues from S184 to S201 which are believed to form the transmembrane segment IV were substituted individually with cysteine and labeled with a thiol-selective nitroxide reagent. Most of the labeled mutants exhibited significant oxoglutarate transport in reconstituted liposomes, where they were examined by EPR as a function of the incident microwave power in the presence and absence of two paramagnetic perturbants, i.e., the hydrophobic molecular oxygen or the hydrophilic chromium oxalate complex. The periodicity of the sequence-specific variation in the spin-label mobility and the O(2) accessibility parameters unambiguously identifies the fourth transmembrane segment of the mitochondrial oxoglutarate carrier as an alpha-helix. The accessibility to chromium oxalate is out of phase with oxygen accessibility, indicating that the helix is amphipatic, with the hydrophilic face containing the residues found to be important for transport activity by site-directed mutagenesis and chemical modification. The helix is strongly packed, as indicated by the values of normalized mobility, which also suggest that the conformational changes occurring during transport probably involve the N-terminal region of the helix. PMID- 12731892 TI - Effect of hydroxylamine on photosystem II: reinvestigation of electron paramagnetic resonance characteristics reveals possible S state intermediates. AB - Previous work in many laboratories has established that hydroxylamine reduces the S(1) state of the water oxidizing complex (WOC) in one-electron steps. Significant levels of what can now be defined as the S(-1)* state are achieved by specific (concentration and incubation length) hydroxylamine treatments. This state has already been studied by electron paramagnetic resonance spectrometry (EPR), and unusual EPR signals were noted (for example, see Sivaraja, M., and Dismukes, G. C. (1988) Biochemistry 27, 3467-3475). We have now reinvestigated these initial experiments and confirmed many of the original observations. We then utilized more recent EPR markers for the S(0) and S(1) states to further explore the S(-1)* state. The broad radical "split" type EPR signal, produced by 200 K illumination of samples prepared to give a high yield of the S(-1)* state, is shown to most likely reflect a trapped intermediate state between S(-1)* and S(0)*, since samples where this signal is present can be warmed in the dark to produce S(0)*. The threshold for advancement from S(-1)* to S(0)* is near 200 K, as the yield of broad radical decreases and S(0)* multiline EPR signal increases with length of 200 K illumination. Advancement of S(0)* to S(1) is limited at 200 K, but S(1) can be restored by 273 K illumination. Illumination of these hydroxylamine-treated samples at temperatures below 77 K gives a second broad radical EPR signal. The line shape, decay, and other properties of this new radical signal suggest that it may arise from an interaction in the S(-2)* or lower S states, which are probably present in low yield in these samples. Illumination below 20 K of S(0)* state samples containing methanol, and therefore exhibiting the S(0) multiline signal, gives rise to a third broad radical with distinctive line shape. The characteristics of the three broad radicals are similar to those found from interactions between Y(Z)(*) and other S states. The evidence is presented that they do represent intermediate states in S state turnover. Further work is now needed to identify these radicals. PMID- 12731890 TI - Characterization of glycosylation sites of the epidermal growth factor receptor. AB - The epidermal growth factor receptor is a transmembrane glycoprotein that mediates the cellular responses to epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha). In this study of the human EGF receptor naturally expressed in A431 cells, the glycosylation sites of the full-length, membrane-bound receptor and of a secreted form of the receptor were characterized by mass spectrometry. Our data show that the naturally expressed human EGF receptor is fully glycosylated on eight of the 11 canonical sites; two of the sites are not glycosylated, and one is partially glycosylated, a pattern of site usage similar but not identical to those reported for the recombinant human EGF receptor heterologously expressed in Chinese hamster ovary cells. We also confirm the partial glycosylation of an atypical NNC site first identified in the receptor expressed in Chinese hamster ovary cells. We show that an additional canonical site in the secreted form of the receptor is fully glycosylated. While the pattern of glycosylation is the same for the sites shared by the full-length and the secreted forms of the receptor, the oligosaccharides of the full-length receptor are more extensively processed. Finally, we provide evidence that in addition to the known secreted form of the receptor, a proteolytic cleavage product of the receptor corresponding to the full extracytoplasmic, ligand binding domain is present in the conditioned medium. PMID- 12731893 TI - Regulation of manganese uptake in Synechocystis 6803 by RfrA, a member of a novel family of proteins containing a repeated five-residues domain. AB - The rfrA gene was identified in a suppressor screen of a Synechocystis sp. PCC 6803 strain deficient in both mntC, encoding a component of an ABC transport system for manganese, and psbO, encoding the extrinsic manganese stabilizing protein of photosystem II (PSII). A spontaneous suppressor mutant (DeltaCDeltaO rfrA-Sup) has a point mutation in rfrA, which restores photosynthetic activity to the DeltamntCDeltapsbO double mutant. Manganese transport and photosynthesis are related in that manganese is essential to the function of PSII, and the state of cellular manganese availability influences the rate of oxygen evolution mediated by PSII. Oxygen evolution experiments with the DeltaCDeltaO rfrA-Sup mutant revealed that the mechanism of suppression is not through a direct modification of PSII. Instead, radioactive manganese uptake experiments indicated that RfrA is a regulator of a high affinity manganese transport system different from the more thoroughly characterized manganese ABC transport system in Synechocystis 6803. RfrA was named for the repeated five-residues domain in the amino terminus of the protein. The RFR domain defines a 16-member family in Synechocystis 6803. Predicted proteins with RFR domains have also been identified in other organisms, but RfrA is the first member of this family to be linked to a physiological process. PMID- 12731894 TI - Modulation of the interactions of isolated ryanodine receptors of rabbit skeletal muscle by Na+ and K+. AB - Ryanodine receptors (RyRs) of skeletal muscle, as calcium release channels, have been found to form semicrystalline arrays in the membrane of sarcoplasmic reticulum. Recently, both experimental observations and theoretical simulations suggested cooperative coupling within interlocking RyRs. To better understand the interactions between RyRs and their modulation, the aggregation and dissociation of isolated RyRs in aqueous medium containing various Na(+) and K(+) concentrations were investigated using photon correlation spectroscopy (PCS) and atomic force microscopy (AFM). RyRs aggregated readily at low salt concentrations. However, a different behavior was observed in the presence of Na(+) or K(+). Detectable aggregates were formed in 5 microg/mL RyR sample when the concentration of Na(+) and K(+) was reduced from 1 M to below 0.28 and 0.23 M, respectively. The dissociation of RyR aggregates was also examined when raising the salt concentration. While aggregates formed in 0.15 M NaCl medium could reverse almost completely, those formed in 0.15 M KCl medium only dissolved partly. When keeping the total salt concentration at 0.15 M, the aggregation and dissociation of RyRs were seen to evidently depend on the relative concentration of Na(+) and K(+). The interaction between RyRs was strengthened with increasing Na(+)/K(+) ratios in the mixed medium. Accompanying this, a decrease of [(3)H]ryanodine binding occurred. The results obtained with PCS and AFM provide further evidence for the interaction between RyRs and suggest the importance of Na(+), K(+), and their relative composition in modulating the interaction and cooperation between RyRs in vivo. PMID- 12731895 TI - Role of hydration in the closed-to-open transition involved in Ca2+ binding by troponin C. AB - Troponin C (TnC) is the Ca(2+)-binding subunit of the troponin complex of vertebrate skeletal muscle. It consists of two structurally homologous domains, N and C, connected by an exposed alpha-helix. The C-domain has two high-affinity sites for Ca(2+) that also bind Mg(2+), whereas the N-domain has two low-affinity sites for Ca(2+). Previous studies using isolated N- and C-domains showed that the C-domain apo form was less stable than the N-domain. Here we analyzed the stability of isolated N-domain (F29W/N-domain) against urea and pressure denaturation in the absence and in the presence of glycerol using fluorescence spectroscopy. Increasing the glycerol concentration promoted an increase in the stability of the protein to urea (0-8 M) in the absence of Ca(2+). Furthermore, the ability to expose hydrophobic surfaces normally promoted by Ca(2+) binding or low temperature under pressure was partially lost in the presence of 20% (v/v) glycerol. Glycerol also led to a decrease in the Ca(2+) affinity of the N-domain in solution. From the ln K(obs) versus ln a(H)2(O), we obtained the number of water molecules (63.5 +/- 8.7) involved in the transition N <=>N:Ca(2) that corresponds to an increase in the exposed surface area of 571.5 +/- 78.3 A(2). In skinned fibers, the affinity for Ca(2+) was also reduced by glycerol, although the effect was much less pronounced than in solution. Our results demonstrate quantitatively that the stability of this protein and its affinity for Ca(2+) are critically dependent on protein hydration. PMID- 12731896 TI - Conformational coupling of Mg2+ and Ca2+ on the three-state folding of calexcitin B. AB - Calexcitin (CE) is a calcium sensor protein that has been implicated in associative learning through the Ca(2+)-dependent inhibition of K(+) channels and activation of ryanodine receptors. CE(B), the major CE variant, was identified as a member of the sarcoplasmic Ca(2+) binding protein family: proteins that can bind both Ca(2+) and Mg(2+). We have now determined the intrinsic Ca(2+) and Mg(2+) binding affinities of CE(B) and investigated their interplay on the folding and structure of CE(B). We find that urea denaturation of CE(B) displays a three-state unfolding transition consistent with the presence of two structural domains. Through a combination of spectroscopic and denaturation studies we find that one domain likely possesses molten globule structure and contains a mixed Ca(2+)/Mg(2+) binding site and a Ca(2+) binding site with weak Mg(2+) antagonism. Furthermore, ion binding to the putative molten globule domain induces native structure formation. The other domain contains a single Ca(2+)-specific binding site and has native structure, even in the absence of ion binding. Ca(2+) binding to CE(B) induces the formation of a recessed hydrophobic pocket. On the basis of measured ion binding affinities and intracellular ion concentrations, it appears that Mg(2+)-CE(B) represents the resting state and Ca(2+)-CE(B) corresponds to the active state, under physiological conditions. PMID- 12731897 TI - Pressure-induced fusogenic conformation of vesicular stomatitis virus glycoprotein. AB - Vesicular stomatitis virus (VSV) is composed of a ribonucleoprotein core surrounded by a lipid envelope presenting an integral glycoprotein (G). The homotrimeric VSV G protein exhibits a membrane fusion activity that can be elicited by low pH. The fusion event is crucial to entry into the cell and disassembly followed by viral replication. To understand the conformational changes involved in this process, the effects of high hydrostatic pressure and urea on VSV particles and isolated G protein were investigated. With pressures up to 3.0 kbar VSV particles were converted into the fusogenic conformation, as measured by a fusion assay and by the binding of bis-ANS. The magnitude of the changes was similar to that promoted by lowering the pH. To further understand the relationship between stability and conversion into the fusion-active states, the stability of the G protein was tested against urea and high pressure. High urea produced a large red shift in the tryptophan fluorescence of G protein whereas pressure promoted a smaller change. Pressure induced equal fluorescence changes in isolated G protein and virions, indicating that virus inactivation induced by pressure is due to changes in the G protein. Fluorescence microscopy showed that pressurized particles were capable of fusing with the cell membrane without causing infection. We propose that pressure elicits a conformational change in the G protein, which maintains the fusion properties but suppresses the entry of the virus by endocytosis. Binding of bis-ANS indicates the presence of hydrophobic cavities in the G protein. Pressure also caused an increase in light scattering of VSV G protein, reinforcing the hypothesis that high pressure elicits the fusogenic activity of VSV G protein. This "fusion-intermediate state" induced by pressure has minor changes in secondary structure and is likely the cause of nonproductive infections. PMID- 12731898 TI - Feshbach-resonance-induced atomic filamentation and quantum pair correlation in atom-laser-beam propagation. AB - We study the propagation of an atom laser beam through a spatial region with a magnetic field tuned around a Feshbach resonance. Magnetic fields below the resonance produce an effective focusing Kerr medium that causes a modulational instability of the atomic beam. Under appropriate circumstances, this results in beam breakup and filamentation seeded by quasiparticle fluctuations and in the generation of correlated atomic pairs. PMID- 12731899 TI - Vortex states of rapidly rotating dilute Bose-Einstein condensates. AB - We show that, in the Thomas-Fermi regime, the cores of vortices in rotating dilute Bose-Einstein condensates adjust in radius as the rotation velocity, Omega, grows, thus precluding a phase transition associated with core overlap at high vortex density. In both a harmonic trap and a rotating hard-walled bucket, the core size approaches a limiting fraction of the intervortex spacing. At large rotation speeds, a system confined in a bucket develops, within Thomas-Fermi, a hole along the rotation axis, and eventually makes a transition to a giant vortex state with all the vorticity contained in the hole. PMID- 12731901 TI - Quantum phase diffusions of a spinor condensate. AB - We discuss the quantum phases and their diffusion in a spinor-1 atomic Bose Einstein condensate. For ferromagnetic interactions, we obtain the exact ground state distribution of the phase fluctuations corresponding to the total atom number (N), the magnetization (M), and the alignment (or hypercharge) (Y) of the system. The mean-field ground state is shown to be stable against these fluctuations, which dynamically recover the two continuous symmetries associated with the conservation of N and M as in current experiments. PMID- 12731900 TI - Coreless vortex formation in a spinor Bose-Einstein condensate. AB - Coreless vortices were phase imprinted in a spinor Bose-Einstein condensate. The three-component order parameter of F=1 sodium condensates held in a Ioffe Pritchard magnetic trap was manipulated by adiabatically reducing the magnetic bias field along the trap axis to zero. This distributed the condensate population across its three spin states and created a spin texture. Each spin state acquired a different phase winding which caused the spin components to separate radially. PMID- 12731902 TI - Collective excitations of a trapped Bose-Einstein condensate in the presence of a 1D optical lattice. AB - We study low-lying collective modes of an elongated 87Rb condensate produced in a 3D magnetic harmonic trap with the addition of a 1D periodic potential which is provided by a laser standing wave along the axial direction. While the transverse breathing mode remains unperturbed, quadrupole and dipole oscillations along the optical lattice are strongly modified. Precise measurements of the collective mode frequencies at different heights of the optical barriers provide a stringent test of the theoretical model recently introduced [M. Kramer, Phys. Rev. Lett. 88, 180404 (2002)]]. PMID- 12731903 TI - Towards new tests of strong-field gravity with measurements of surface atomic line redshifts from neutron stars. AB - In contrast to gravity in the weak-field regime, which has been subject to numerous experimental tests, gravity in the strong-field regime is largely unconstrained by observations. We show that gravity theories that pass solar system tests, but that diverge from general relativity in the strong-field regime, predict neutron stars with significantly different properties than their general relativistic counterparts. The range of redshfits of surface atomic lines predicted by such theories is significantly wider than the uncertainty introduced by our lack of knowledge of the equation of state of ultradense matter. Measurements of such lines with x-ray observatories can thus put new constraints on strong-field gravity. PMID- 12731904 TI - Spacelike brane actions. AB - We derive effective actions for "spacelike branes" (S-branes) and find a solution describing the formation of fundamental strings in the rolling tachyon background. The S-brane action is a Dirac-Born-Infeld action for Euclidean world volumes defined in the context of time-dependent tachyon condensation of non-BPS (Bogomol'nyi-Prasad-Sommerfield) branes. It includes gauge fields and, in particular, a scalar field associated with translation along the time direction. We show that the BIon spike solutions constructed in this system correspond to the production of a confined electric flux tube (a fundamental string) at late time of the rolling tachyon. PMID- 12731905 TI - Measurements of the Decay KL-->e+ e- mu+ mu-. AB - The KTeV experiment at Fermilab has isolated a total of 132 events from the rare decay K(L)-->e+ e- mu+ mu-, with an estimated background of 0.8 events. The branching ratio of this mode is determined to be [2.69+/-0.24(stat)+/ 0.12(syst)]x10(-9), with a radiative cutoff of M(2)(ee mu mu)/M(2)(K)>0.95. The first measurement using this mode of the parameter alpha from the D'Ambrosio Isidori-Portoles (DIP) model of the K(L)gamma*gamma* vertex yields a result of 1.59+/-0.37, consistent with values obtained from other decay modes. Because of the limited statistics, no sensitivity is found to the DIP parameter beta. We use this decay mode to set limits on CP and lepton violation. PMID- 12731907 TI - Bd-->phi KS CP asymmetries as an important probe of supersymmetry. AB - The decay B(d)-->phi K(S) is a special probe of physics beyond the standard model (SM), since it has no SM tree level contribution. Motivated by recent data suggesting a deviation from the SM for its time-dependent CP asymmetry, we examine supersymmetric explanations. Chirality preserving contributions are generically small, unless gluino is relatively light. Higgs contributions are also too small to explain a large asymmetry. Chirality flipping LR and RL gluino contributions actually can provide sizable effects without conflict with all related results. We discuss how various insertions can be distinguished, and argue the needed sizes of mass insertions are reasonable. PMID- 12731909 TI - Isospin violation in e+ e- -->BB. AB - The ratio of the B+ B- and B0B0 production rates in e+ e- annihilation is computed as a function of the B meson velocity and BB*pi coupling constant, using a nonrelativistic effective field theory. PMID- 12731910 TI - Measurement of spin-correlation parameters ANN, ASS, and ASL at 2.1 GeV in proton proton elastic scattering. AB - At the Cooler Synchrotron COSY/Julich spin-correlation parameters in elastic proton-proton (pp) scattering have been measured with a 2.11 GeV polarized proton beam and a polarized hydrogen atomic beam target. We report results for A(NN), A(SS), and A(SL) for c.m. scattering angles between 30 degrees and 90 degrees. Our data on A(SS)--the first measurement of this observable above 800 MeV- clearly disagrees with predictions of available pp scattering phase-shift solutions while A(NN) and A(SL) are reproduced reasonably well. We show that in the direct reconstruction of the scattering amplitudes from the body of available pp elastic scattering data at 2.1 GeV the number of possible solutions is considerably reduced. PMID- 12731906 TI - Observation of B0 -->D0K0 and B0 -->D0K*0 decays. AB - We report on a search for B(0)-->D(*0)K(*0) decays based on 85 x 10(6) BB events collected with the Belle detector at KEKB. The B(0)-->D0K(0) and B(0)-->D0K(*0) decays have been observed for the first time with the branching fractions B(B(0)- >D0K(0))=(5.0(+1.3)(-1.2)+/-0.6)x10(-5) and B(B(0)-->D0K(*0))=(4.8(+1.1)(-1.0)+/ 0.5)x10(-5). No significant signal has been found for the B(0)-->D(*0)K*0) and B(0)-->D(*0)K(*0) decay modes, and upper limits at 90% C.L. are presented. PMID- 12731911 TI - Direct decays from superdeformed states in 192Pb observed using time-correlated gamma-ray spectroscopy. AB - Correlations of decays above and below isomeric states in the normally deformed minimum of 192Pb have been used to identify discrete transitions in the decay of the superdeformed (SD) band. The data establish the absolute excitation energy of the lowest observed SD level as 4.425 MeV. Extrapolation to the bandhead indicates that the excitation energy of the superdeformed well in 192Pb is 0.5 MeV lower than in the heavier isotope 194Pb. The results confirm the trend to decreasing excitation energy with decreasing neutron number predicted by both a macroscopic Strutinsky method approach and microscopic mean field calculations. PMID- 12731912 TI - New manifestation of atomic parity violation in cesium: a chiral optical gain induced by linearly polarized 6S-7S excitation. AB - We have detected, by using stimulated emission, an atomic parity violation (APV) in the form of a chiral optical gain of a cesium vapor on the 7S-6P(3/2) transition, consecutive to linearly polarized 6S-7S excitation. We demonstrate the validity of this detection method of APV, by presenting a 9% accurate measurement of expected sign and magnitude. We stress several advantages of this new approach which fully exploits the cylindrical symmetry of the setup. Future measurements at the percent level will provide an important cross-check of an existing more precise result obtained by a different method. PMID- 12731913 TI - Rydberg cold collisions dominated by ultralong range potential. AB - In this work we measure the time evolution of the population resulting from energy-transfer collisions as a function of the energy difference between the entrance and exit collisional channels using a sample of cold Rydberg atoms produced in a rubidium magneto-optical trap. The 34S(1/2) population, produced by collisions between atoms in the 33P(3/2) state, is monitored as a function of time through the pulsed-field ionization technique. The experimental results are compared with a recent published model based on a two-body interaction considering an attractive potential [Phys. Rev. A 65, 023405 (2002)]]; which is calculated according to a recent Letter by Boisseau et al. [Phys. Rev. Lett. 88, 133004 (2002)]]. The agreement is remarkable, which suggests the existence of such ultralong range potential proposed by Boisseau et al. PMID- 12731908 TI - Improved direct measurement of the parity-violation parameter Ab using a mass tag and momentum-weighted track charge. AB - We present an improved direct measurement of the parity-violation parameter A(b) in the Z boson-b-quark coupling using a self-calibrating track-charge technique applied to a sample enriched in Z-->bb events via the topological reconstruction of the B hadron mass. Manipulation of the Stanford Linear Collider electron-beam polarization permits the measurement of A(b) to be made independently of other Z pole coupling parameters. From the 1996-1998 sample of 400,000 hadronic Z decays, produced with an average beam polarization of 73.4%, we find A(b)=0.906+/ 0.022(stat)+/-0.023(syst). PMID- 12731914 TI - Structure, rotational dynamics, and superfluidity of small OCS-doped He clusters. AB - The structural and dynamical properties of carbonyl sulfide (OCS) molecules solvated in helium clusters are studied using reptation quantum Monte Carlo, for cluster sizes n=3-20 He atoms. Computer simulations allow us to establish a relation between the rotational spectrum of the solvated molecule and the structure of the He solvent, and of both with the onset of superfluidity. Our results agree with a recent spectroscopic study of this system and provide a more complex and detailed microscopic picture of this system than inferred from experiments. PMID- 12731915 TI - Multimode Hong-Ou-mandel interference. AB - We consider multimode two-photon interference at a beam splitter by photons created by spontaneous parametric down-conversion. The resulting interference pattern is shown to depend upon the transverse spatial symmetry of the pump beam. In an experiment, we employ the first-order Hermite-Gaussian modes in order to show that, by manipulating the pump beam, one can control the resulting two photon interference behavior. We expect these results to play an important role in the engineering of quantum states of light for use in quantum information processing and quantum imaging. PMID- 12731916 TI - Precision measurement and compensation of optical stark shifts for an ion-trap quantum processor. AB - Using optical Ramsey interferometry, we precisely measure the laser-induced ac Stark shift on the S(1/2)-D(5/2) "quantum bit" transition near 729 nm in a single trapped 40Ca+ ion. We cancel this shift using an additional laser field. This technique is of particular importance for the implementation of quantum information processing with cold trapped ions. As a simple application we measure the atomic phase evolution during a n x 2 pi rotation of the quantum bit. PMID- 12731917 TI - Dynamical Janssen effect on granular packing with moving walls. AB - Apparent mass (M(app)) measurements at the bottom of granular packings inside a vertical tube in relative motion are reported. They demonstrate that Janssen's model is valid over a broad range of velocities v. The variability of the measurements is lower than for static packings and the theoretical exponential increase of M(app) with the height of the packing is precisely followed (the corresponding characteristic screening length is of the order of the tube diameter). The limiting apparent mass at large heights is independent of v and significantly lower than the static value. PMID- 12731918 TI - Dense, rapid flows of inelastic grains under gravity. AB - The standard hydrodynamic description does not apply to the rapid flow regime of inelastic grains in the dense limit. Emphasizing the role of inelastic loss and collapse, we propose a new approach relying on a nonlocal dissipation scheme. Our model succeeds in accounting qualitatively and quantitatively for the linear profile of velocity found in experiments on dense gravity-driven flows. PMID- 12731919 TI - Surface roughness and hydrodynamic boundary slip of a newtonian fluid in a completely wetting system. AB - The influence of surface roughness on the boundary condition for the flow of a Newtonian fluid near a hard wall has been investigated by measurement of the hydrodynamic drainage force. The degree of slip is found to increase with surface roughness. This leads to the conclusion that in most practical situations boundary slip takes place, leading to a reduction of the drainage force. PMID- 12731920 TI - Controlling the fiber diameter during electrospinning. AB - We present a simple analytical model for the forces that determine jet diameter during electrospinning as a function of surface tension, flow rate, and electric current in the jet. The model predicts the existence of a terminal jet diameter, beyond which further thinning of the jet due to growth of the whipping instability does not occur. Experimental data for various electrospun fibers attest to the accuracy of the model. PMID- 12731921 TI - Laser-induced hydrodynamic instability of fluid interfaces. AB - We report on a new class of electromagnetically driven fluid interface instability. Using the optical radiation pressure of a cw laser to bend a very soft near-critical liquid-liquid interface, we show that it becomes unstable for sufficiently large beam power P, leading to the formation of a stationary beam centered liquid microjet. We explore the behavior of the instability onset by tuning the interface softness with temperature and varying the size of the exciting beam. The instability mechanism is experimentally demonstrated. It simply relies on total reflection of light at the deformed interface whose condition provides the universal scaling relation for the onset P(S) of the instability. PMID- 12731922 TI - Kinetic roughening in two-phase fluid flow through a random Hele-Shaw cell. AB - A nonlocal interface equation is derived for two-phase fluid flow, with arbitrary wettability and viscosity contrast, c=(mu(1)-mu(2))/(mu(1)+mu(2)), in a model porous medium defined as a Hele-Shaw cell with random gap b(0)+delta b. Fluctuations of both capillary and viscous pressure are explicitly related to the microscopic quenched disorder, yielding conserved, nonconserved, and power-law correlated noise terms. Two length scales are identified that control the possible scaling regimes and which scale with capillary number Ca as l(1) approximately b(0)(cCa)(-1/2) and l(2) approximately b(0)Ca-1. Exponents for forced fluid invasion are obtained from numerical simulation and compared with recent experiments. PMID- 12731923 TI - Ordered and disordered patterns in two-phase flows in microchannels. AB - We show that wetting properties crucially control the patterns in two-phase flows of immiscible fluids in microchannels. Ordered patterns, continuously entrained by the flow, are obtained when one phase completely wets the walls, while disordered patterns, intermittently adhering to the channel walls, are unavoidably produced when wetting is partial. A lower limit for the channel sizes capable of generating well structured objects (drops, pears, pearl necklaces, ...) is presented. PMID- 12731924 TI - Experimental studies of the Bohm criterion in a two-ion-species plasma using laser-induced fluorescence. AB - The Bohm criterion is studied experimentally in the case of a two ion species plasma. Measurements are carried out in Ar and Ar+He plasmas (PA(r I) approximately 0.1 mtorr, 0< or =P(He)/P(Ar)< or =25, and 0< or =n(+)(He)/n(e)< or =0.5, T(e)< or =2 eV) created in an unmagnetized dc hot filament discharge confined by surface multidipole magnetic fields. Laser-induced fluorescence (LIF) measurements of Ar II ion velocity distribution functions (ivdfs) within the presheath up to the sheath edge show that the ions reach the sheath edge traveling faster than their individual Bohm speed by more than 75%, approaching a speed equal to the ion sound speed of the system. PMID- 12731925 TI - Nucleation of single-walled carbon nanotubes. AB - The nucleation pathway for single-wall carbon nanotubes on a metal surface is demonstrated by a series of total energy calculations using density functional theory. Incorporation of pentagons at an early stage of nucleation is energetically favorable as they reduce the number of dangling bonds and facilitate curvature of the structure and bonding to the metal. In the presence of the metal surface, nucleation of a closed cap or a capped single-wall carbon nanotube is overwhelmingly favored compared to any structure with dangling bonds or to a fullerene. PMID- 12731926 TI - Refraction and reflection of diffusion fronts. AB - Diffusion waves form the basis of several measurement technologies in materials science as well as in biological systems. They are, however, so heavily damped that their observation is a real challenge to the experimentalist. We show that accurate information about the refraction-like and reflection-like behavior of diffusion waves can be obtained by studying diffusion fronts. For this we use hydrogen in a metal as a model system and visualize its 2D migration with an optical indicator. The similarities between classical optics and diffusion, in particular, the applicability of Snell's law to diffusive systems are discussed. Our measurements are in good agreement with numerical simulations. PMID- 12731927 TI - Pressure-induced crossover between diffusive and displacive mechanisms of phase transitions in single-crystalline alpha-GeO2. AB - We present the first example of the phase transition occurring via the different kinetic mechanisms, displacive or diffusive, competing with each other in quartz like alpha-GeO2 single crystals. Upon room-pressure heating, alpha-GeO2 transforms to the rutile-type phase (the alpha-->r transition) via the diffusive mechanism. With increase of the treatment pressure the diffusive mode of the temperature-induced alpha-->r transition is substituted at approximately 4 GPa by a displacive-like mode, and then at approximately 6 GPa the transition type changes from the alpha-->r sequence to a displacive martensitic-like transition to a distorted rutile-like phase (alpha-->r'. A crossover between diffusive and displacive transition modes suggests a new way to control the meso- and nanometer scale morphology of high-pressure phases. PMID- 12731928 TI - Structural changes in block copolymers: coupling of electric field and mobile ions. AB - We argue that the presence of dissociated ions in block copolymers under electric fields can induce strong morphological changes and even lead to phase transitions. We investigate, in particular, diblock copolymers in the body centered cubic (bcc) phase. In pure dielectric materials (no free charges), a dielectric breakdown is expected to occur for large enough electric fields, preempting any structural phase transition. On the other hand, dissociated ions are predicted to induce a phase transition to a hexagonal array of cylinders, at fields of about 10 V/microm or even lower. The strength of this mechanism can be tuned by controlling the amount of free ions present. PMID- 12731929 TI - Nearest-neighbor configuration in (GaIn)(NAs) probed by x-ray absorption spectroscopy. AB - Ga(1-x)In(x)N(y)As(1-y) is a promising material system for the fabrication of inexpensive "last-mile" optoelectronic components. However, details of its atomic arrangement and the relationship to observed optical properties is not fully known. Particularly, a blueshift of emission wavelength is observed after annealing. In this work, we use x-ray absorption fine structure to study the chemical environment around N atoms in the material before and after annealing. We find that as-grown molecular beam epitaxy material consists of a nearly random distribution of atoms, while postannealed material shows segregation of In toward N. Ab initio simulations show that this short-range ordering creates a more thermodynamically stable alloy and is responsible for blueshifting the emission. PMID- 12731930 TI - Role of rigidity in the fluid-solid transition. AB - We examine the fluid-solid transition for a hard-disk system. By counting the near neighbors in the average configurations of a grand-canonical Monte Carlo simulation, this enables us to relate the thermodynamic transition with the rigidity theory, since we find that the coordination number in the fluid-solid transition is close to the coordination number predicted by a mean field rigidity theory, due to dynamical jamming of particles, where the contact region between disks is the radial ring outside a disk with a maximum allowed coordination number that is not bigger than six. Using these ideas, we were able to produce a continuous glass-like transition when nucleation of rigidity is suppressed. PMID- 12731931 TI - Equilibrium model of bimodal distributions of epitaxial island growth. AB - We present a nanostructure diagram for use in designing heteroepitaxial systems of quantum dots. The nanostructure diagram is computed using a new equilibrium statistical physics model and predicts the island size and shape distributions for a range of combinations of growth temperature and amount of deposited material. The model is applied to Ge on Si(001), the archetype for bimodal island growth, and the results compare well with data from atomic force microscopy of Ge/Si islands grown by chemical vapor deposition. PMID- 12731932 TI - Effect of C60 molecular rotation on nanotribology. AB - The effect of C60 molecular rotation on the nanotribological properties of C60 single crystal surfaces has been studied by atomic/frictional force microscopy. The orientational order-disorder phase transition, in which the high temperature C60 free rotation is reduced to a low temperature hindered rotation, is shown to give rise to an abrupt change in friction and adhesion. This change in frictional force is quantitatively consistent with the observed change in adhesion. The similar slopes of the friction versus load curves in both phases indicate that the friction coefficient in the two phases remains about the same. Hence the C60 rotation does not provide an additional energy dissipation channel in the friction process. PMID- 12731934 TI - Density functionals not based on the electron gas: local-density approximation for a Luttinger liquid. AB - By shifting the reference system for the local-density approximation (LDA) from the electron gas to other model systems, one obtains a new class of density functionals, which by design account for the correlations present in the chosen reference system. This strategy is illustrated by constructing an explicit LDA for the one-dimensional Hubbard model. While the traditional ab initio LDA is based on a Fermi liquid (the three-dimensional interacting electron gas), this one is based on a Luttinger liquid. First applications to inhomogeneous Hubbard models, including one containing a localized impurity, are reported. PMID- 12731933 TI - Unusual magnetic state in lithium-doped MoS2 nanotubes. AB - We report on the very peculiar magnetic properties of an ensemble of very weakly coupled lithium-doped MoS2 nanotubes. The magnetic susceptibility chi of the system is nearly 3 orders of magnitude greater than in typical Pauli metals, yet there is no evidence for any instability which would alleviate this highly frustrated state. Instead, the material exhibits peculiar paramagnetic stability down to very low temperatures, with no evidence of a quantum critical point as T- >0 in spite of clear evidence for strongly correlated electron behavior. The exceptionally weak intertube interactions appear to lead to a realization of a near-ideal one-dimensional state in which fluctuations prevent the system from reordering magnetically or structurally. PMID- 12731935 TI - Localization length in anderson insulator with Kondo impurities. AB - The localization length, xi, in a two-dimensional Anderson insulator depends on the electron spin scattering rate by magnetic impurities, tau(-1)(s). For antiferromagnetic sign of the exchange, the time tau(s) is itself a function of xi, due to the Kondo correlations. We demonstrate that the unitary regime of localization is impossible when the concentration of magnetic impurities, n(M), is smaller than a critical value, n(c). For n(M)>n(c), the dependence of xi on the dimensionless conductance, g, is reentrant, crossing over to unitary, and back to orthogonal behavior upon increasing g. Sensitivity of Kondo correlations to a weak parallel magnetic field results in a giant parallel magnetoresistance. PMID- 12731936 TI - One-dimensional confinement and enhanced Jahn-Teller instability in LaVO3. AB - Ordering and quantum fluctuations of orbital degrees of freedom are studied theoretically for LaVO3 in the spin-C-type antiferromagnetic state. The effective Hamiltonian for the orbital pseudospin shows strong one-dimensional anisotropy due to the negative interference among various exchange processes. This significantly enhances the instability toward lattice distortions for the realistic estimate of the Jahn-Teller coupling by first-principle LDA+U calculations, instead of favoring the orbital singlet formation. This explains well the experimental results on the anisotropic optical spectra as well as the proximity of the two transition temperatures for spin and orbital orderings. PMID- 12731937 TI - Nonballistic spin-field-effect transistor. AB - We propose a spin-field-effect transistor based on spin-orbit coupling of both the Rashba and the Dresselhaus types. Different from earlier proposals, spin transport through our device is tolerant against spin-independent scattering processes. Hence the requirement of strictly ballistic transport can be relaxed. This follows from a unique interplay between the Dresselhaus and the Rashba coupling; these can be tuned to have equal strengths, leading to k-independent eigenspinors even in two dimensions. We discuss two-dimensional devices as well as quantum wires. In the latter, our setup presents strictly parabolic dispersions which avoids complications from anticrossings of different bands. PMID- 12731938 TI - First-order phase transition from the vortex liquid to an amorphous solid. AB - We present a systematic study of the topology of the vortex solid phase in superconducting Bi2Sr2CaCu2O8 samples with low doses of columnar defects. A new state of vortex matter imposed by the presence of geometrical contours associated with the random distribution of columns is found. The results show that the first order liquid-solid transition in this vortex matter does not require a structural symmetry change. PMID- 12731939 TI - Tail states in clean superconductors with magnetic impurities. AB - We analyze the behavior of the density of states in a singlet s-wave superconductor with weak magnetic impurities in the clean limit. By using the method of optimal fluctuation and treating the order parameter self-consistently we show that the density of states is finite everywhere in the superconducting gap, and that it varies as ln(N(E) proportional to -/E-Delta(0)/((7-d)/4) near the mean field gap edge Delta(0) in a d-dimensional superconductor. In contrast to most studied cases the optimal fluctuation is strongly anisotropic. PMID- 12731940 TI - Stable vortex-antivortex molecules in mesoscopic superconducting triangles. AB - A thermodynamically stable vortex-antivortex pattern has been revealed in equilateral mesoscopic type I superconducting triangles, contrary to type II superconductors where similar patterns are unstable. The stable vortex-antivortex "molecule" appears due to the interplay between two factors: a repulsive vortex antivortex interaction in type I superconductors and the vortex confinement in the mesoscopic triangle. PMID- 12731941 TI - Charge freezing in the zigzag chain PrBa2Cu4O8 cuprate. AB - We report nuclear quadrupole resonance (NQR) studies on the chain Cu sites of PrBa2Cu4O8, a quasi-one-dimensional conductor with a nearly quarter-filled band. The nuclear spin-lattice relaxation rate 1/T1 shows a pronounced peak near 100 K caused by fluctuations of electric field gradient. Similar peak was observed for the spin-echo decay rate 1/T2, however, at a different temperature near 50 K. These results and broadening of the NQR spectrum at low temperatures indicate that slow charge fluctuations of either electronic or ionic origin freeze gradually at low temperatures. PMID- 12731942 TI - Intrinsic tunneling spectra of Bi2(Sr(2-x)Lax)CuO(6+delta). AB - We have measured intrinsic-tunneling spectra of a single CuO-layer La-doped Bi2(Sr(2-x)Lax)CuO(6+delta) (Bi2201-Lax). Despite a difference of a factor of 3 in the optimal superconducting critical temperatures for Bi2201-La0.4 and Bi2212 (32 and 95 K, respectively) and different spectral energy scales, we find that the pseudogap vanishes at a similar characteristic temperature T* approximately 230-300 K for both compounds. We find also that, in Bi2201-Lax, pseudogap humps are seen as sharp peaks and, in fact, even dominate the intrinsic spectra. PMID- 12731943 TI - Oscillatory exchange bias due to an antiferromagnet with incommensurate spin density waves. AB - Oscillatory exchange bias in both magnitude and in sign has been observed in epitaxial (100)Cr/Ni(81)Fe19 bilayers due to the incommensurate spin-density waves in antiferromagnetic (100)Cr layers. Salient effects due to the spin-flip transition between longitudinal and transverse spin-density waves as well as that of expanding wavelength have been observed. PMID- 12731944 TI - High-energy spin dynamics in La1.69Sr0.31NiO4. AB - To test the prediction that the dispersion of the magnetic resonance in superconducting YBa2Cu3O(6+x) is similar to magnons in an incommensurate antiferromagnet, we have mapped out the spin dynamics in a stripe-ordered nickelate, La(2-x)SrxNiO4, with x approximately equal to 0.31, using inelastic neutron scattering. We observe spin-wave excitations up to 80 meV emerging from the incommensurate magnetic peaks with a surprisingly large and almost isotropic spin velocity: variant Planck's over 2 pi c(s) approximately 0.32 eV A. A comparison indicates that the inferred spin-excitation spectrum is not, by itself, an adequate model for the magnetic resonance feature of the superconductor. PMID- 12731945 TI - Frustrated spin model as a hard-sphere liquid. AB - We show that one-dimensional topological objects (kinks) are natural degrees of freedom for an antiferromagnetic Ising model on a triangular lattice. Its ground states and the coexistence of spin ordering with an extensive zero-temperature entropy can easily be understood in terms of kinks forming a hard-sphere liquid. Using this picture we explain effects of quantum spin dynamics on that frustrated model, which we also study numerically. PMID- 12731946 TI - Singlet excitations in pyrochlore: a study of quantum frustration. AB - We apply the contractor renormalization (CORE) method to the spin half Heisenberg antiferromagnet on the frustrated checkerboard and pyrochlore lattices. Their ground states are spin-gapped singlets which break lattice symmetry. Their effective Hamiltonians describe fluctuations of orthogonal singlet pairs on tetrahedral blocks, at an emergent low energy scale. We discuss low temperature thermodynamics and new interpretations of finite size numerical data. We argue that our results are common to many models of quantum frustration. PMID- 12731947 TI - Quantum computing with spatially delocalized qubits. AB - We analyze the operation of quantum gates for neutral atoms with qubits that are delocalized in space, i.e., the computational basis states are defined by the presence of a neutral atom in the ground state of one out of two trapping potentials. The implementation of single-qubit gates as well as a controlled phase gate between two qubits is discussed and explicit calculations are presented for rubidium atoms in optical microtraps. Furthermore, we show how multiqubit highly entangled states can be created in this scheme. PMID- 12731948 TI - Evaluation of holonomic quantum computation: adiabatic versus nonadiabatic. AB - Based on the analytical solution to the time-dependent Schrodinger equations, we evaluate the holonomic quantum computation beyond the adiabatic limit. Besides providing rigorous confirmation of the geometrical prediction of holonomies, the present dynamical resolution offers also a practical means to study the nonadiabaticity induced effects for the universal qubit operations. PMID- 12731949 TI - Interplay between geometry and flow distribution in an airway tree. AB - Uniform flow distribution in a symmetric volume can be realized through a symmetric branched tree. It is shown here, however, by 3D numerical simulation of the Navier-Stokes equations, that the flow partitioning can be highly sensitive to deviations from exact symmetry if inertial effects are present. The flow asymmetry is quantified and found to depend on the Reynolds number. Moreover, for a given Reynolds number, we show that the flow distribution depends on the aspect ratio of the branching elements as well as their angular arrangement. Our results indicate that physiological variability should be severely restricted in order to ensure adequate fluid distribution through a tree. PMID- 12731950 TI - Wave initiation through spatiotemporally controllable perturbations. AB - We study the initiation of pulses and fronts in a two-dimensional catalytic reaction-diffusion system: CO oxidation on Pt(110). Using a computer-controlled mobile focused laser beam, we impart various patterns (in space and time) of localized temperature "kicks" to the surface. We explore, and also rationalize through modeling, the cooperativity of such individually subcritical perturbations in both the excitable and the bistable regime. PMID- 12731951 TI - Oscillatory dispersion and coexisting stable pulse trains in an excitable medium. AB - For planar wave trains in excitable media, we found a novel type of anomalous dispersion distinguished by bistable domains in the dependence of the propagation velocity on the wavelength. Within one medium alternative stable pulse trains can coexist having the same wavelength but different velocities. The phenomenon is related to oscillatory recovery of excitations, which causes small amplitude oscillations in the refractory tail of pulses. Crucial for the bistability is that the pulses in the trains are locked into one oscillation maximum in the tail of the preceding pulse in the train. PMID- 12731952 TI - Modularity and extreme edges of the internet. AB - We study the spectral properties of a diffusion process taking place on the Internet network focusing on the slowest decaying modes. These modes identify an underlying modular structure roughly corresponding to individual countries. For instance, in the slowest decaying mode the diffusion current flows from Russia to U.S. military sites. Quantitatively the modular structure manifests itself in a 10 times larger participation ratio of its slow decaying modes compared to a random scale-free network. We propose to use the fraction of nodes participating in slow decaying modes as a general measure of the modularity of a network. For the 100 slowest decaying modes of the Internet this fraction is approximately 30%. Finally, we suggest that the degree of isolation of an individual module can be assessed by comparing its participation in different diffusion modes. PMID- 12731953 TI - Random vibrational networks and the renormalization group. AB - We consider the properties of vibrational dynamics on random networks, with random masses and spring constants. The localization properties of the eigenstates contrast greatly with the Laplacian case on these networks. We introduce several real-space renormalization techniques which can be used to describe this dynamics on general networks, drawing on strong disorder techniques developed for regular lattices. The renormalization group is capable of elucidating the localization properties, and provides, even for specific network instances, a fast approximation technique for determining the spectra which compares well with exact results. PMID- 12731954 TI - Comment on "Off-diagonal long-range order in Bose liquids: irrotational flow and quantization of circulation". PMID- 12731956 TI - Comment on "Nonmonotonic d(x2-y2) superconducting order parameter in Nd(2 x)CexCuO4". PMID- 12731958 TI - Comment on "Direct determination of interfacial magnetic moments with a magnetic phase transition in Co nanoclusters on Au111". PMID- 12731960 TI - Collisional decoherence observed in matter wave interferometry. AB - We study the loss of spatial coherence in the extended wave function of fullerenes due to collisions with background gases. From the gradual suppression of quantum interference with increasing gas pressure we are able to support quantitatively both the predictions of decoherence theory and our picture of the interaction process. We thus explore the practical limits of matter wave interferometry at finite gas pressures and estimate the required experimental vacuum conditions for interferometry with even larger objects. PMID- 12731961 TI - Geometric phase in open systems. AB - We calculate the geometric phase associated with the evolution of a system subjected to decoherence through a quantum-jump approach. The method is general and can be applied to many different physical systems. As examples, two main sources of decoherence are considered: dephasing and spontaneous decay. We show that the geometric phase is completely insensitive to the former, i.e., it is independent of the number of jumps determined by the dephasing operator. PMID- 12731962 TI - Forces between conducting surfaces due to spatial variations of surface potential. AB - We describe analytical and numerical methods for calculating forces between conductors due to variations of electrostatic surface potential across their surfaces. In the simple case where the spatial variation of surface potential gives rise to uniform power spectra, we show that the electrostatic force can be large in comparison with, and scale in approximately the same way with distance of closest approach as, the Casimir force. Patch potentials that are consistent with existing experimental data could give rise to forces with a magnitude of 4% of the Casimir force at separations of 0.1 microm. PMID- 12731963 TI - New features of the thermal Casimir force at small separations. AB - The difference of the thermal Casimir forces at different temperatures between real metals is shown to increase with a decrease of the separation distance. This opens new opportunities for the demonstration of the thermal dependence of the Casimir force. Both configurations of two parallel plates and a sphere above a plate are considered. Different approaches to the theoretical description of the thermal Casimir force are shown to lead to different measurable predictions. PMID- 12731964 TI - Amplification of local instabilities in a Bose-Einstein condensate with attractive interactions. AB - We study the collapse of large homogeneous Bose-Einstein condensates due to intrinsic attractive interactions. We observe the amplification of a local instability by seeding a momentum state p and suddenly switching the scattering length negative via a Feshbach resonance. We also observe the appearance of atoms in the conjugate momentum state as required by momentum conservation. For large condensates, the time scale for this depletion process becomes faster than that for global collapse. PMID- 12731965 TI - Atoms and molecules in lattices: Bose-Einstein condensates built on a shared vacuum. AB - In optical lattices where each site is occupied in its lowest energy state by a superposition of zero, one, and two atoms, one can in a controllable manner convert the atomic pair into a molecule while retaining the vacuum and one-atom amplitudes. The microscopic quantum coherence on each site between the vacuum and the single molecule component leads to a macroscopically populated molecular condensate when the lattice is removed. PMID- 12731966 TI - Matter-wave gap solitons in atomic band-gap structures. AB - We demonstrate that a Bose-Einstein condensate in an optical lattice forms a reconfigurable matter-wave structure with a band-gap spectrum, which resembles a nonlinear photonic crystal for light waves. We study in detail the case of a two dimensional square optical lattice and show that this atomic band-gap structure allows nonlinear localization of atomic Bloch waves in the form of two dimensional matter-wave gap solitons. PMID- 12731967 TI - All-versus-nothing violation of local realism for two entangled photons. AB - It is shown that the Greenberger-Horne-Zeilinger theorem can be generalized to the case with only two entangled particles. The reasoning makes use of two photons which are maximally entangled both in polarization and in spatial degrees of freedom. In contrast to Cabello's argument of "all versus nothing" nonlocality with four photons [Phys. Rev. Lett. 87, 010403 (2001)]], our proposal to test the theorem can be implemented with linear optics and thus is well within the reach of current experimental technology. PMID- 12731968 TI - Spatial structure of a vortex in low density neutron matter. AB - We study in a fully self-consistent approach the structure of a vortex in low density superfluid neutron matter. We determine that the matter density profile of a vortex shows a significant depletion in the region of the core, a feature never reported for a vortex state in a Fermi superfluid. PMID- 12731969 TI - d-dimensional black hole entropy spectrum from quasinormal modes. AB - Starting from recent observations about quasinormal modes, we use semiclassical arguments to derive the Bekenstein-Hawking entropy spectrum for d-dimensional spherically symmetric black holes. We find that, as first suggested by Bekenstein, the entropy spectrum is equally spaced: S(BH)=kln((m(0))n, where m(0) is a fixed integer that must be derived from the microscopic theory. As shown in O. Dreyer, gr-qc/0211076, 4D loop quantum gravity yields precisely such a spectrum with m(0)=3 providing the Immirzi parameter is chosen appropriately. For d-dimensional black holes of radius R(H)(M), our analysis predicts the existence of a unique quasinormal mode frequency in the large damping limit omega((d))(M)=alpha((d))c/R(H)(M) with coefficient [formula: see text], where m(0) is an integer. PMID- 12731970 TI - Fine structure of the QCD string spectrum. AB - Using advanced lattice methods in quantum chromodynamics, three distinct scales are established in the excitation spectrum of the gluon field around a static quark-antiquark pair as the color source separation R is varied. On the shortest length scale, the excitations are consistent with states created by local gluon field operators arising from a multipole operator product expansion. An intermediate crossover region below 2 fm is identified with a dramatic rearrangement of the level orderings. On the largest length scale of 2-3 fm, the spectrum agrees with that expected for stringlike excitations. The energies nearly reproduce asymptotic pi/R string gaps, but exhibit a fine structure, providing important clues for developing an effective bosonic string description. PMID- 12731971 TI - e+ e- annihilation into J/psi + J/psi. AB - Recent measurements by the Belle Collaboration of the exclusive production of two charmonia in e(+)e(-) annihilation differ substantially from theoretical predictions. We suggest that a significant part of the discrepancy can be explained by the process e(+)e(-)-->J/psi+J/psi. Because the J/psi+J/psi production process can proceed through fragmentation of two virtual photons into two cc pairs, its cross section may be larger than that for J/psi+eta(c) by about a factor of 3.7, in spite of a suppression factor alpha(2)/alpha(2)(s) that is associated with the QED and QCD coupling constants. PMID- 12731973 TI - Effects of symmetry energy on two-nucleon correlation functions in heavy-ion collisions induced by neutron-rich nuclei. AB - Using an isospin-dependent transport model, we study the effects of nuclear symmetry energy on two-nucleon correlation functions in heavy-ion collisions induced by neutron-rich nuclei. We find that the density dependence of the nuclear symmetry energy affects significantly the nucleon emission times in these collisions, leading to larger values of two-nucleon correlation functions for a symmetry energy that has a stronger density dependence. Two-nucleon correlation functions are thus useful tools for extracting information about the nuclear symmetry energy from heavy-ion collisions. PMID- 12731972 TI - 21Na(p,gamma)22Mg reaction and oxygen-neon novae. AB - The 21Na(p,gamma)22Mg reaction is expected to play an important role in the nucleosynthesis of 22Na in oxygen-neon novae. The decay of 22Na leads to the emission of a characteristic 1.275 MeV gamma-ray line. This report provides the first direct measurement of the rate of this reaction using a radioactive 21Na beam, and discusses its astrophysical implications. The energy of the important state was measured to be E(c.m.)=205.7+/-0.5 keV with a resonance strength omegagamma=1.03+/-0.16(stat)+/-0.14(sys) meV. PMID- 12731974 TI - Asymptotic properties of self-energy coefficients. AB - We investigate the asymptotic properties of higher-order binding corrections to the one-loop self-energy of excited states in atomic hydrogen. We evaluate the historically problematic A60 coefficient for all P states with principal quantum numbers n40 T. We argue that this reduction does not arise from 4f polarization but reflects renormalization of the quasiparticle states by the field. PMID- 12731988 TI - Electrical detection of spin accumulation in a p-type GaAs quantum well. AB - We report on experiments in which a spin-polarized current is injected from a GaMnAs ferromagnetic electrode into a GaAs layer through an AlAs barrier. The resulting spin polarization in GaAs is detected by measuring how the tunneling current, to a second GaMnAs ferromagnetic electrode, depends on the orientation of its magnetization. Our results can be accounted for by sequential tunneling with the nonrelaxed spin splitting of the chemical potential, that is, spin accumulation, in GaAs. We discuss the conditions on the hole spin relaxation time in GaAs that are required to obtain the large effects we observe. PMID- 12731989 TI - Interaction-driven spin precession in quantum-dot spin valves. AB - We analyze spin-dependent transport through spin valves composed of an interacting quantum dot coupled to two ferromagnetic leads. The spin on the quantum dot and the linear conductance as a function of the relative angle theta of the leads' magnetization directions is derived to lowest order in the dot-lead coupling strength. Because of the applied bias voltage spin accumulates on the quantum dot, which for finite charging energy experiences a torque, resulting in spin precession. The latter leads to a nontrivial, interaction-dependent, theta dependence of the conductance. In particular, we find that the spin-valve effect is reduced for all theta not equal pi. PMID- 12731990 TI - Confined Shockley surface states on the (111) facets of gold clusters. AB - Combining low-temperature scanning tunneling microscopy and spectroscopy with high-resolution ultraviolet photoemission, we have revealed a confined Shockley surface state on the (111) facets of gold clusters with about N=10(4) atoms grown in nanopits on highly oriented graphite. With tunneling spectroscopy, we observed energy dependent nodal patterns in the dI/dV maps, which are in quantitative agreement with the two-dimensional confinement of the surface state within the hexagonal facet area. The results indicate that the lattice of the ionic cores influences the electronic properties of the clusters significantly. PMID- 12731991 TI - Current detection of superradiance and induced entanglement of double quantum dot excitons. AB - We propose to measure the superradiance effect by observing the current through a semiconductor double-dot system. An electron and a hole are injected separately into one of the quantum dots to form an exciton and then recombine radiatively. We find that the stationary current shows oscillatory behavior as one varies the interdot distance. The amplitude of oscillation can be increased by incorporating the system into a microcavity. Furthermore, the current is suppressed if the dot distance is small compared to the wavelength of the emitted photon. This photon trapping phenomenon generates the entangled state and may be used to control the emission of single photons at predetermined times. PMID- 12731992 TI - Spin-entangled currents created by a triple quantum dot. AB - We propose a simple setup of three coupled quantum dots in the Coulomb blockade regime as a source for spatially separated currents of spin-entangled electrons. The entanglement originates from the singlet ground state of a quantum dot with an even number of electrons. To preserve the entanglement of the electron pair during its extraction to the drain leads, the electrons are transported through secondary dots. This prevents one-electron transport by energy mismatch, while joint transport is resonantly enhanced by conservation of the total two-electron energy. PMID- 12731993 TI - Phase sensitive experiments in ferromagnetic-based Josephson junctions. AB - We have measured the ground state of ferromagnetic Josephson junctions using a single dc SQUID (superconducting quantum interference device).We show that the Josephson coupling is either positive (0 coupling) or negative (pi coupling) depending on the ferromagnetic layer thickness. As expected, the sign change of the Josephson coupling is observed as a shift of half a quantum flux in the SQUID diffraction pattern when operating in the linear limit. PMID- 12731994 TI - Proximity of the layered organic conductors alpha-(BEDT-TTF)2MHg(SCN)4 (M = K, NH4) to a charge-ordering transition. AB - While the optical properties of the superconducting salt alpha-(BEDT-TTF)(2) NH4Hg(SCN)(4) remain metallic down to 2 K, in the nonsuperconducting K analog a pseudogap develops at frequencies of about 200 cm(-1) for temperatures T<200 K. We show that the optical conductivity calculated with exact-diagonalization techniques on an extended Hubbard model at quarter filling is consistent with the observed low-frequency feature. We argue that the different optical responses observed are a consequence of the proximity of these compounds to a charge ordering transition driven by the intermolecular Coulomb repulsion. PMID- 12731995 TI - Circuit theory of unconventional superconductor junctions. AB - We extend the circuit theory of superconductivity to cover transport and proximity effect in mesoscopic systems that contain unconventional superconductor junctions. The approach fully accounts for zero-energy Andreev bound states forming at the surface of unconventional superconductors. As a simple application, we investigate the transport properties of a diffusive normal metal in series with a d-wave superconductor junction. We reveal the competition between the formation of Andreev bound states and proximity effect that depends on the crystal orientation of the junction interface. PMID- 12731996 TI - Theory of the vortex matter transformations in high-Tc superconductor YBCO. AB - Flux line lattice in type II superconductors undergoes a transition into a "disordered" phase such as vortex liquid or vortex glass, due to thermal fluctuations and random quenched disorder. We quantitatively describe the competition between the thermal fluctuations and the disorder using the Ginzburg Landau approach. The following T-H phase diagram of YBCO emerges. There are just two distinct thermodynamical phases, the homogeneous and the crystalline one, separated by a single first order transition line. The line, however, makes a wiggle near the experimentally claimed critical point at 12 T. The "critical point" is reinterpreted as a (noncritical) Kauzmann point in which the latent heat vanishes and the line is parallel to the T axis. The magnetization, the entropy, and the specific heat discontinuities at melting compare well with experiments. PMID- 12731997 TI - Ferromagnetic superconductivity driven by changing Fermi surface topology. AB - We introduce a simple but powerful zero temperature Stoner model to explain the unusual phase dia-gram of the ferromagnetic superconductor, UGe2. Triplet superconductivity is driven in the ferromagnetic phase by tuning the majority spin Fermi level through one of two peaks in the paramagnetic density of states (DOS). Each peak is associated with a metamagnetic jump in magnetization. The twin-peak DOS may be derived from a tight-binding, quasi-one-dimensional band structure, inspired by previous band-structure calculations. PMID- 12731998 TI - Strong superconductivity with local Jahn-Teller phonons in C60 solids. AB - We analyze fulleride superconductivity at experimental doping levels, treating the electron-electron and electron-phonon interactions on an equal footing, and demonstrate that the Jahn-Teller phonons create a local (intramolecular) pairing which is surprisingly resistant to the Coulomb repulsion, despite the weakness of retardation in these low-bandwidth systems. The requirement for coherence throughout the solid then yields a very strong doping dependence to T(c), one consistent with experiment and much stronger than expected from standard Eliashberg theory. PMID- 12731999 TI - Charge-order-induced sharp Raman peak in Sr14Cu24O41. AB - In the two-leg S=1/2 ladders of Sr14Cu24O41, a modulation of the exchange coupling arises from the charge order within the other structural element, the CuO2 chains. In general, breaking transla-tional invariance by modulation causes gaps within the dispersion of elementary excitations. We show that the gap induced by the charge order can drastically change the magnetic Raman spectrum, leading to the sharp peak observed in Sr14Cu24O41. This sharp Raman line gives insight into the charge-order periodicity and hence into the distribution of carriers. The much broader spectrum of La6Ca8Cu24O41 reflects the response of an undoped ladder in the absence of charge order. PMID- 12732000 TI - Low-dimensional spin S = 1/2 system at the quantum critical limit: Na2V3O7. AB - We report the results of measurements of the dc susceptibility and the 23Na-NMR response of Na2V3O7, a recently synthesized, nonmetallic low dimensional spin system. Our results indicate that, upon reducing the temperature to below 100 K, the V4+ moments are gradually quenched, leaving only one moment out of nine active. The NMR data reveal a phase transition at very low temperatures. With decreasing applied field H, the critical temperature shifts towards T=0 K, suggesting that Na2V3O7 may be regarded as an insulator reaching a quantum critical point at H=0. PMID- 12732001 TI - Intrinsic nonlinear ferromagnetic relaxation in thin metallic films. AB - We propose an intrinsic mechanism for ferromagnetic relaxation in thin films that can dominate competing linear mechanisms even for rapidly relaxing metals. In particular, we use an analytic theory of four-magnon scattering to demonstrate rapid decay for technologically important systems involving high moment materials subject to large rotations. A micromagnetic simulation is used to verify the results. PMID- 12732002 TI - Magnetization transport and quantized spin conductance. AB - We analyze transport of magnetization in insulating systems described by a spin Hamiltonian. The magnetization current through a quasi-one-dimensional magnetic wire of finite length suspended between two bulk magnets is determined by the spin conductance which remains finite in the ballistic limit due to contact resistance. For ferromagnetic systems, magnetization transport can be viewed as transmission of magnons, and the spin conductance depends on the temperature T. For antiferromagnetic isotropic spin-1/2 chains, the spin conductance is quantized in units of order (gmu(B))(2)/h at T=0. Magnetization currents produce an electric field and, hence, can be measured directly. For magnetization transport in electric fields, phenomena analogous to the Hall effect emerge. PMID- 12732003 TI - Detection of XY behavior in weakly anisotropic quantum antiferromagnets on the square lattice. AB - We consider the Heisenberg antiferromagnet on the square lattice with S=1/2 and very weak easy-plane exchange anisotropy; by means of the quantum Monte Carlo method, based on the continuous-time loop algorithm, we find that the thermodynamics of the model is highly sensitive to the presence of tiny anisotropies and is characterized by a crossover between isotropic and planar behavior. We discuss the mechanism underlying the crossover phenomenon and show that it occurs at a temperature which is characteristic of the model. The expected Berezinskii-Kosterlitz-Thouless transition is observed below the crossover: a finite range of temperatures consequently opens for experimental detection of noncritical 2D XY behavior. Direct comparison is made with uniform susceptibility data relative to the S=1/2 layered antiferromagnet Sr2CuO2Cl2. PMID- 12732005 TI - Theory of highly directional emission from a single subwavelength aperture surrounded by surface corrugations. AB - We present a theoretical foundation for the beaming of light displayed by a single subwavelength aperture in an appropriately corrugated metal film [H. J. Lezec, Science 297, 820 (2002)]]. Good agreement is found between calculations and experimental data. We show that beaming is due to the formation of electromagnetic surface resonances and that the beam direction, width, and wavelength at which it occurs can be selected by tuning geometrical parameters of the structure. PMID- 12732004 TI - Magnetic domain structure and magnetic anisotropy in Ga1-xMn(x)As. AB - Large, well-defined magnetic domains, on the scale of hundreds of micrometers, are observed in Ga1-xMn(x)As epilayers using a high-resolution magneto-optical imaging technique. The orientations of the magnetic moments in the domains clearly show in-plane magnetic anisotropy, which changes through a second-order transition from a biaxial mode (easy axes nearly along [100] and [010]) at low temperatures to an unusual uniaxial mode (easy axis along [110]) as the temperature increases above about T(c)/2. This transition is a result of the interplay between the natural cubic anisotropy of the GaMnAs zinc-blende structure and a uniaxial anisotropy which attribute to the effects of surface reconstruction. PMID- 12732006 TI - Influence of synthesis conditions on the structural and optical properties of passivated silicon nanoclusters. AB - First-principles molecular dynamics and quantum Monte Carlo techniques are employed to gain insight into the effect of preparation conditions on the structural and optical properties of silicon nanoparticles. Our results demonstrate that (i) kinetically limited nanostructures form different core structures than bulk-derived crystalline clusters, (ii) the type of core structure that forms depends on how the cluster is passivated during synthesis, and (iii) good agreement with measured optical gaps can be obtained for nanoparticles with core structures different from those derived from the bulk. PMID- 12732007 TI - Reversible valence tautomerism induced by a single-shot laser pulse in a cobalt iron Prussian blue analog. AB - Reversible valence tautomeric conversion induced by a single-shot laser pulse (8 ns duration) with a photon excitation energy of 2.38 eV has been observed in Na0.36Co1.32Fe(CN)(6).5.6H(2)O. A photoswitching process with accompanying magnetization and color changes was successfully achieved within the pulse duration at high temperature (above 200 K) in a thermal hysteresis loop. This unusual photoeffect originates from an optical charge transfer between Fe and Co atoms and evolves due to a cooperative interaction among the local photoexcited sites. PMID- 12732008 TI - Breakdown transients in ultrathin gate oxides: transition in the degradation rate. AB - We report a sharp threshold at 4 V in the growth rate of breakdown spots in thin films of SiO2 on silicon. This provides some of the first information concerning the electronic structure of the breakdown spot. PMID- 12732009 TI - Measuring quantum entanglement without prior state reconstruction. AB - It is shown that, despite strong nonlinearity, entanglement of formation of a two qubit state can be measured without prior state reconstruction. Collective measurements on a small number of copies are provided that allow one to determine quantum concurrence via estimation of only four parameters. It is also pointed out that another entanglement measure based on so-called "negativity" can also be measured in a similar way. The result is related to the general problem: What kind of information can be extracted efficiently from an unknown quantum state? PMID- 12732010 TI - Power of entanglement in quantum communication. AB - A communication channel is a physical system that transfers information from one place to another. Examples of communication channels include wires, optical fibers, and chains of spins that propagate spin waves through a medium. This Letter shows that the power-limited communication capacity of a multimode optical fiber or a set of parallel spin chains can be enhanced by introducing nonlinear couplings between the modes or chains. In particular, M coupled, entangled modes can send M bits in the same time it takes a single mode to send a single bit, and in the same time it takes M uncoupled, unentangled modes to send sqrt[M] bits. PMID- 12732011 TI - Experimental demonstration of tripartite entanglement and controlled dense coding for continuous variables. AB - A tripartite entangled state of bright optical field is experimentally produced using an Einstein-Podolsky-Rosen entangled state for continuous variables and linear optics. The controlled dense coding among a sender, a receiver, and a controller is demonstrated by exploiting the tripartite entanglement. The obtained three-mode "position" correlation and relative "momentum" correlation between the sender and the receiver, and thus the improvements of the measured signal to noise ratios of amplitude and phase signals with respect to the shot noise limit are 3.28 and 3.18 dB, respectively. If the mean photon number n equals 11 the channel capacity can be controllably inverted between 2.91 and 3.14. When n is larger than 1.0 and 10.52, the channel capacity of the controlled dense coding is predicted to exceed the ideal single channel capacity of coherent and squeezed state light communication, respectively. PMID- 12732012 TI - Unconditionally secure key distribution based on two nonorthogonal states. AB - We prove the unconditional security of the Bennett 1992 protocol, by using a reduction to an entanglement distillation protocol initiated by a local filtering process. The bit errors and the phase errors are correlated after the filtering, and we can bound the amount of phase errors from the observed bit errors by an estimation method involving nonorthogonal measurements. The angle between the two states shows a trade-off between accuracy of the estimation and robustness to noises. PMID- 12732013 TI - Entanglement measure for composite systems. AB - A general description of entanglement is suggested as an action realized by an arbitrary operator over given disentangled states. The related entanglement measure is defined. Because of its generality, this definition can be employed for any physical systems, pure or mixed, equilibrium or nonequilibrium, and characterized by any type of operators, whether these are statistical operators, field operators, spin operators, or anything else. Entanglement of any number of parts from their total ensemble forming a multiparticle composite system can be determined. Interplay between entanglement and ordering, occurring under phase transitions, is analyzed by invoking the concept of operator order indices. PMID- 12732014 TI - Exceeding the classical capacity limit in a quantum optical channel. AB - The amount of information transmissible through a communications channel is determined by the noise characteristics of the channel and by the quantities of available transmission resources. In classical information theory, the amount of transmissible information can be increased twice at most when the transmission resource is doubled for fixed noise characteristics. In quantum information theory, however, the amount of information transmitted can increase even more than twice. We present a proof-of-principle demonstration of this superadditivity of classical capacity of a quantum channel by using the ternary symmetric states of a single photon, and by event selection from a weak coherent light source. We also show how the superadditive coding gain, even in a small code length, can boost the communication performance of the conventional coding technique. PMID- 12732015 TI - Formation of electrically active clusterized neural networks. AB - Ordinarily, in vitro neurons self-organize into homogeneous networks of single neurons linked by dendrites and axons. We show that under special conditions they can also self-organize into neuronal clusters, which are linked by bundles of axons. Multielectrode array measurement reveals that the clusterized networks are also electrically active and exhibit synchronized bursting events similar to those observed in the homogeneous networks. From time-lapse recording, we deduced the features required for the neuronal clusterized versus homogeneous self organization and developed a simple model for testing their validity. PMID- 12732016 TI - Motion of an adhesive gel in a swelling gradient: a mechanism for cell locomotion. AB - Motivated by the motion of nematode sperm cells, we present a model for the motion of an adhesive gel on a solid substrate. The gel polymerizes at the leading edge and depolymerizes at the rear. The motion results from a competition between a self-generated swelling gradient and the adhesion on the substrate. The resulting stress provokes the rupture of the adhesion points and allows for the motion. The model predicts an unusual force-velocity relation which depends in significant ways on the point of application of the force. PMID- 12732017 TI - Fast chain contraction during protein folding: "foldability" and collapse dynamics. AB - Theory indicates that at least some proteins will undergo a rapid and unimpeded collapse, like a disorganized hydrophobic chain, prior to folding. Yet experiments continue to find signs of an organized, or barrier-limited, collapse in even the fastest (approximately mus) folding proteins. Does the kinetic barrier represent a signature of the equilibrium "foldability" of these molecules? We have measured the rate of chain contraction in two nonfolding analogs of a very fast-collapsing protein. We find that these chains contract on the same time scale (approximately 10(-5)s) as the natural protein, and both pass over an energetic barrier at least as large as that encountered by the protein. The equilibrium foldability of the native structure therefore does not alone determine the dynamics of collapse; even the disordered chains contract approximately 1000x slower than expected for an ideal chain. PMID- 12732018 TI - Universal elasticity and fluctuations of nematic gels. AB - We study elasticity of spontaneously orientationally ordered amorphous solids, characterized by a vanishing transverse shear modulus, as realized by nematic elastomers and gels. We show that local heterogeneities and elastic nonlinearities conspire to lead to anomalous nonlocal universal elasticity controlled by a nontrivial infrared fixed point. Namely, such solids are characterized by universal shear and bending moduli that, respectively, vanish and diverge at long scales, are universally incompressible, and exhibit a universal negative Poisson ratio and a non-Hookean elasticity down to arbitrarily low strains. Based on expansion about five dimensions, we argue that the nematic order is stable to thermal fluctuation and local heterogeneities down to d(lc)<3. PMID- 12732019 TI - Flow-induced twist-compression in a twisted nematic cell. AB - An optical convergent-beam guided-wave technique is used to explore in detail the dynamic flow effects in a twisted nematic cell. During switch-on it is found that the dynamic flow compresses the director twist to regions close to the cell walls. For high fields this twist compression takes the cell far beyond the Mauguin limit and it no longer effectively guides the polarization of the light through the cell. This results in a very fast switch to a transient dark state. PMID- 12732020 TI - Mixing and condensation in a wet granular medium. AB - We have studied the effect of small amounts of added liquid on the dynamic behavior of a granular system consisting of a mixture of glass beads of two different sizes. Segregation of the large beads to the top of the sample is found to depend in a nontrivial way on the liquid content. A transition to viscoplastic behavior occurs at a critical liquid content, which depends upon the bead size. We show that this transition can be interpreted as a condensation due to the hysteretic liquid bridge forces connecting the beads, and we provide the corresponding phase diagram. PMID- 12732021 TI - Probing states in the Mott insulator regime in the case of coherent bosons trapped in an optical lattice. AB - We propose a method to probe states in the Mott insulator regime produced from a condensate in an optical lattice. We consider a system in which we create time dependent number fluctuations in a given site by turning off the atomic interactions and lowering the potential barriers on a nearly pure Mott state to allow the atoms to tunnel between sites. We calculate the expected interference pattern and number fluctuations from such a system and show that one can potentially observe a deviation from a pure Mott state. We also discuss a method in which to detect these number fluctuations using time-of-flight imaging. PMID- 12732022 TI - From Cooper pairs to Luttinger liquids with bosonic atoms in optical lattices. AB - We propose a scheme to observe strongly correlated fermionic phenomena with bosonic atoms in optical lattices. For different values of the sign and strength of the scattering lengths, it is possible to reach either a "superconducting" regime, where the system exhibits atomic pairing, or a Luttinger liquid behavior. We identify the range of parameters where these phenomena appear, illustrate our predictions with numerical calculations, and show how to detect the presence of pairing. PMID- 12732024 TI - Testing the stability of fundamental constants with the 199Hg+ single-ion optical clock. AB - Over a two-year duration, we have compared the frequency of the 199Hg+ 5d(10)6s (2)S(1/2)(F=0)<-->5d(9)6s(2) (2)D(5/2)(F=2) electric-quadrupole transition at 282 nm with the frequency of the ground-state hyperfine splitting in neutral 133Cs. These measurements show that any fractional time variation of the ratio nu(Cs)/nu(Hg) between the two frequencies is smaller than +/-7 x 10(-15) yr(-1) (1sigma uncertainty). According to recent atomic structure calculations, this sets an upper limit to a possible fractional time variation of g(Cs)(m(e)/m(p))alpha(6.0) at the same level. PMID- 12732023 TI - Search for variations of fundamental constants using atomic fountain clocks. AB - Over five years, we have compared the hyperfine frequencies of 133Cs and 87Rb atoms in their electronic ground state using several laser-cooled 133Cs and 87Rb atomic fountains with an accuracy of approximately 10(-15). These measurements set a stringent upper bound to a possible fractional time variation of the ratio between the two frequencies: d/dt ln([(nu(Rb))/(nu(Cs))]=(0.2+/-7.0)x 10(-16) yr( 1) (1sigma uncertainty). The same limit applies to a possible variation of the quantity (mu(Rb)/mu(Cs))alpha(-0.44), which involves the ratio of nuclear magnetic moments and the fine structure constant. PMID- 12732025 TI - New experimental constraints on non-Newtonian forces below 100 microm. AB - We have searched for large deviations from Newtonian gravity by means of a finite frequency microcantilever-based experiment. Our data eliminate from consideration mechanisms of deviation that posit strengths approximately 10(4) times Newtonian gravity at length scales of 20 microm. This measurement is 3 orders of magnitude more sensitive than others that provide constraints at similar length scales. PMID- 12732026 TI - Inflationary spacetimes are incomplete in past directions. AB - Many inflating spacetimes are likely to violate the weak energy condition, a key assumption of singularity theorems. Here we offer a simple kinematical argument, requiring no energy condition, that a cosmological model which is inflating--or just expanding sufficiently fast--must be incomplete in null and timelike past directions. Specifically, we obtain a bound on the integral of the Hubble parameter over a past-directed timelike or null geodesic. Thus inflationary models require physics other than inflation to describe the past boundary of the inflating region of spacetime. PMID- 12732027 TI - Relating a gluon mass scale to an infrared fixed point in pure gauge QCD. AB - We show that in pure gauge QCD (or any pure non-Abelian gauge theory) the condition for the existence of a global minimum of energy with a gluon (gauge boson) mass scale also implies the existence of a fixed point of the beta function. We argue that the frozen value of the coupling constant found in some solutions of the Schwinger-Dyson equations of QCD can be related to this fixed point. We also discuss how the inclusion of fermions modifies this property. PMID- 12732028 TI - Saturation of nuclear matter and short-range correlations. AB - A fully self-consistent treatment of short-range correlations in nuclear matter is presented. Different implementations of the determination of the nucleon spectral functions for different interactions are shown to be consistent with each other. The resulting saturation densities are closer to the empirical result when compared with (continuous choice) Brueckner-Hartree-Fock values. Arguments for the dominance of short-range correlations in determining the nuclear matter saturation density are presented. A further survey of the role of long-range correlations suggests that the inclusion of pionic contributions to ring diagrams in nuclear matter leads to higher saturation densities than empirically observed. A possible resolution of the nuclear matter saturation problem is suggested. PMID- 12732030 TI - Entanglement between an electron and a nuclear spin 1/2. AB - We report on the preparation and detection of entangled states between an electron spin 1/2 and a nuclear spin 1/2 in a molecular single crystal. These were created by applying pulses at ESR (9.5 GHz) and NMR (21 MHz, 46 MHz) frequencies. Entanglement was detected by using a special entanglement detector sequence based on a unitary back transformation including phase rotation. PMID- 12732029 TI - Signature for vibrational to rotational evolution along the yrast line. AB - We present a simple method for discerning the evolution from vibrational to rotational structure in nuclei as a function of spin. The prescription is applied to the yrast cascades in the A approximately 110 region and a clear transition from vibrational to rotational motion is found. PMID- 12732031 TI - Measurement of vacuum-assisted photoionization at 1 GeV for Au and Ag targets. AB - We report a measurement of photon impact ionization of K and L shell of Au and K shell of Ag targets in the 1-GeV energy range. We show that the cross section is dominated by a contribution from a new channel called vacuum-assisted photoionization. In this process the energy-momentum balance associated with the removal of the innershell electron is obtained by conversion of a high-energy photon into an electron-positron pair. This measurement is consistent with the theoretical prediction that vacuum-assisted photoionization is the most probable ionization mechanism at very high energies. PMID- 12732032 TI - Auger electron emission from fixed-in-space CO. AB - We have measured the angular distribution of carbon K-Auger electrons from fixed in space, core-ionized, CO molecules in coincidence with the kinetic energy release of the C+ and O+ fragments. We find a very narrow ejection of Auger electrons in the direction of the oxygen and an oscillatory diffraction pattern. Even for similar electron energies, the angular distribution strongly depends on the symmetry of the final state. PMID- 12732034 TI - Doppler-free resonant Raman auger spectroscopy of Ne+ 2s2p(5)3p excited states. AB - Using very high resolution achieved by the Doppler-free resonant Raman Auger technique, we have resolved the lowest terms of the series of inner-valence excitations 2s2p(5)((1,3)P)np 2S, 2P, and 2D in Ne+. The measured Auger anisotropic parameters and branching ratios help to establish the assignments of these levels. The measured lifetime widths are in reasonable agreements with ab initio calculations available in the literature. PMID- 12732033 TI - Experimental determination of the lifetime for the 2p3d(1P0) helium doubly excited state. AB - Two recent theoretical studies [C. Liu, Phys. Rev. A 64, 010501 (2001)]; M. Zitnik, ibid. 65, 032520 (2002)]] predict that the fluorescence lifetimes of helium doubly excited states converging to He+ N=2 should be longer than that of the He+ 2p ion state. This effect is caused by the electric field of the outer electron which, through Stark mixing, gives the inner fluorescing electron some series specific, stabilizing 2s character. We have obtained the first experimental evidence that confirms this effect by measuring the lifetime of the 2p3d(1P0) doubly excited state. This was determined to be 190+/-30 ps compared to 100 ps for the He+ 2p ion state. The measurements were performed using short pulses of synchrotron radiation to form doubly excited states and recording the arrival time of photons from fluorescence. PMID- 12732035 TI - Ultrahot electron formation under excess electron drift through solid Xe. AB - The formation of "hot" (8 eV) electrons under excess electron drift in a moderate electrostatic field through solid xenon has been experimentally proved by observation of secondary electrons emitted from the photocathode. At T=77 K and U=1000 V one drifting electron produces about 20 (172 nm) photons, the efficiency of electric field-to-vacuum ultraviolet emission conversion is 15% tending to grow with temperature. A self-sustained electric discharge has been generated in solid Xe using a three-electrode cell with a zinc cathode. PMID- 12732036 TI - Coulombic energy transfer and triple ionization in clusters. AB - Using neon and its dimer as a specific example, it is shown that excited Auger decay channels that are electronically stable in the isolated monomer can relax in a cluster by electron emission. The decay mechanism, leading to the formation of a tricationic cluster, is based on an efficient energy-transfer process from the excited, dicationic monomer to a neighbor. The decay is ultrafast and expected to be relevant to numerous physical phenomena involving core holes in clusters and other forms of spatially extended atomic and molecular matter. PMID- 12732037 TI - Postpulse molecular alignment measured by a weak field polarization technique. AB - We report a direct nonintrusive observation of alignment and planar delocalization of CO2 after an intense linearly polarized femtosecond laser pulse excitation. The effects are measured by a polarization technique involving a perturbative probe that itself does not induce appreciable alignment. We show that this technique allows one to measure a signal proportional to 1/3, with theta the angle between the molecular axis and the laser polarization. Simulations that support this analysis allow one to characterize the experimentally observed alignment and planar delocalization quantitatively. PMID- 12732038 TI - Scaling properties of simple limiter control. AB - "Simple limiter control" of chaotic systems is analytically and numerically investigated, proceeding from the one-dimensional case to higher dimensions. The properties of the control method are fully described by the one-parameter one dimensional flat-top map family, implying that orbits are stabilized in exponential time, independent of the periodicity and without the need for targeting. Fine-tuning of the control is limited by superexponential scaling in the control space, where orbits of the uncontrolled system are obtained for a set of zero Lebesgue measure. In higher dimensions, simple limiter control is a highly efficient control method, provided that the proper limiter form and placement are chosen. PMID- 12732039 TI - Low-dimensional models for vertically falling viscous films. AB - Long wave evolution on free falling viscous films is described using a new evolution equation. The scaling proposed here brings in the viscous and pressure correction terms that are missing in the existing long-wave equations. Small amplitude expansion of the equation gives a dissipative form of the Kuromoto Sivashinsky equation. Improved accuracy of the new equation over existing equations is demonstrated by comparison of neutral curves with Orr-Sommerfeld equations and experimental data. PMID- 12732040 TI - Anomalous resistivity resulting from MeV-electron transport in overdense plasma. AB - Laser produced hot electron transport in an overdense plasma is studied by three dimensional particle-in-cell simulations. Hot electron currents into the plasma generate neutralizing return currents in the cold plasma electrons, leading to a configuration which is unstable to electromagnetic Weibel and tearing instabilities. The resulting current filaments self-organize through a coalescence process finally settling into a single global current channel. The plasma return current experiences a strong anomalous resistivity due to diffusive flow of cold electrons in the magnetic perturbations. The resulting electrostatic field leads to an anomalously rapid stopping of fast MeV electrons (almost 3 orders of magnitude stronger than that through classical collisional effects). PMID- 12732041 TI - Particle pinch with fully noninductive lower hybrid current drive in Tore Supra. AB - Recently, plasmas exceeding 4 min have been obtained with lower hybrid current drive (LHCD) in Tore Supra. These LHCD plasmas extend for over 80 times the resistive current diffusion time with zero loop voltage. Under such unique conditions the neoclassical particle pinch driven by the toroidal electric field vanishes. Nevertheless, the density profile remains peaked for more than 4 min. For the first time, the existence of an inward particle pinch in steady-state plasma without toroidal electric field, much larger than the value predicted by the collisional neoclassical theory, is experimentally demonstrated. PMID- 12732042 TI - Direct observation of stimulated-Brillouin-scattering detuning by a velocity gradient. AB - We report the first direct evidence of detuning of stimulated Brillouin scattering (SBS) by a velocity gradient, which was achieved by directly measuring the frequency shift of the SBS-driven acoustic wave relative to the local resonant acoustic frequency. We show that in the expanding part of the plasma, ion-acoustic waves are driven off resonance which leads to the saturation of the SBS instability. These measurements are well reproduced by fluid simulations that include the measured flow. PMID- 12732043 TI - Experimental observations of mode-converted ion cyclotron waves in a tokamak plasma by phase contrast imaging. AB - The process of mode conversion, whereby an externally launched electromagnetic wave converts into a shorter wavelength mode(s) in a thermal plasma near a resonance in the index of refraction, is particularly important in a multi-ion species plasma near the ion cyclotron frequency. Using phase contrast imaging techniques (PCI), mode-converted electromagnetic ion cyclotron waves have been detected for the first time in the Alcator C-Mod tokamak near the H-3He ion-ion hybrid resonance region during high power rf heating experiments. The results agree with theoretical predictions. PMID- 12732044 TI - Hydrodynamics of pair-annihilating disclinations in SmC films. AB - The pair annihilation of smectic c-director defects with winding numbers +/-1 in a freestanding SmC film as a representative of the XY model is studied numerically, considering a full coupling of orientational degrees of freedom and hydrodynamics. A reduction of the annihilation time compared to the nonhydrodynamic treatment is observed. It is demonstrated that the +1 disclination moves considerably faster than the -1 one primarily due to hydrodynamic flow, weakly assisted also by elastic anisotropy. The stress tensor terms and material parameters relevant for this effect are identified. PMID- 12732045 TI - Definition and properties of ideal amorphous solids. AB - It is proposed that two ideal amorphous structures, type I and type II, based on maximally random jammed packing of spheres of equal size, form a distinct class of ideal amorphous solids. The ideal amorphous structures contain wide variations in local density, limited by the condition of solidity. Four distinct characteristics, based on statistical geometry and topology, are shown to define this class. Voronoi tessellations carried out on simulated cells of random packed spheres and amorphous polymers give a broad distribution of individual volumes, skewed, with a tail at the high volume end. PMID- 12732046 TI - High-pressure Raman study of Ba doped silicon clathrate. AB - Vibrational properties of Ba doped Si clathrate are investigated at high pressures up to 20 GPa by Raman spectroscopy. Vibrations related with Ba encaged in the Si cages are observed below 100 cm(-1) by low-frequency Raman measurements of Ba8Si46 and Ba6.6Si46 clathrates. The high-pressure Raman spectra obtained for both compression and decompression processes reveal a new phase transition at 7 GPa, and the reversibility of the phase transition at 15 GPa. We investigate the mechanisms of these phase transitions, the interaction between the guest and host frame, and the volume dependence of the vibrational modes. PMID- 12732047 TI - Carbon onions: carriers of the 217.5 nm interstellar absorption feature. AB - Ultraviolet-visible absorption measurements of high purity and well separated carbon onion samples are reported. The results show that, after purification, absorption features from carbon onions match well with the interstellar UV spectrum. The measurements show that the absorption peak position remains constant at 4.55+/-0.1 microm(-1), and the width varies from 1.2-1.6 microm(-1), a key feature of the interstellar spectrum. The similarities between the experimental and observed absorption spectra indicate that carbon onions are very strong candidates for the origin of the UV interstellar absorption peak at 4.6 microm(-1). PMID- 12732048 TI - Responsive solids from cross-linked block copolymers. AB - Novel responsive solids were prepared by randomly cross-linking the polyisoprene chains of a disordered polystyrene-polyisoprene block copolymer. Our experiments show that block copolymer chains, composed of hundreds of repeat units, can undergo reversible order-disorder transitions despite the quenched randomness that arises due to the attachment of one of the blocks to a cross-linked network. The structure, properties, and phase behavior of these materials are determined by a delicate interplay between the density of cross-links and the nature of the ordered state. PMID- 12732050 TI - Mixed alkali effect in glasses. AB - We have applied the bond-valence technique to reverse Monte Carlo produced structural models of mixed alkali phosphate glasses in order to elucidate the mixed alkali effect (MAE) in glasses. For the first time, the MAE is reproduced and understood directly from structural models in quantitative agreement with available experimental results. The two types of alkali ions are randomly mixed and have distinctly different conduction pathways of low dimensionality. This implies that A ions tend to block the pathways for the B ions and vice versa, and this is the main reason for the MAE. PMID- 12732049 TI - Elastic properties of 2D colloidal crystals from video microscopy. AB - Elastic constants of two-dimensional (2D) colloidal crystals are determined by measuring strain fluctuations induced by Brownian motion of particles. Paramagnetic colloids confined to an air-water interface of a pendant drop are crystallized under the action of a magnetic field, which is applied perpendicular to the 2D layer. Using video microscopy and digital image processing we measure fluctuations of the microscopic strain obtained from random displacements of the colloidal particles from their mean (reference) positions. From these we calculate system-size dependent elastic constants, which are extrapolated using finite-size scaling to obtain their values in the thermodynamic limit. The data are found to agree rather well with zero-temperature calculations. PMID- 12732052 TI - Fluorine in silicon: diffusion, trapping, and precipitation. AB - The effect of vacancies on the behavior of F in crystalline Si has been elucidated experimentally for the first time. With positron annihilation spectroscopy and secondary ion mass spectroscopy, we find that F retards recombination between vacancies (V) and interstitials (I) because V and I trap F to form complexes. F diffuses in the V-rich region via a vacancy mechanism with an activation energy of 2.12+/-0.08 eV. After a long annealing time at 700 degrees C, F precipitates have been observed by cross-section transmission electron microscopy which are developed from the V-type defects around the implantation range and the I-type defects at the end of range. PMID- 12732051 TI - Crystal-amorphous polymer interface studied by neutron and x-ray scattering on labeled binary ultralong alkanes. AB - The recently discovered stable semicrystalline phase in binary ultralong alkanes is studied by small-angle neutron and x-ray scattering. Either the shorter or the longer alkane has (CD2)11CD3 ends. It is confirmed that chains are tilted in the crystal layer and found that ca. seven end carbons of the shorter alkane are amorphous. It is also established that the longer chains protrude from the crystal at both ends and that their end groups are preferentially located in the middle of the amorphous layer, suggesting gradual dissipation of orientational order of the chains exiting the crystal. PMID- 12732053 TI - Dopant-pair structures segregated on a hydrogen-terminated Si(100) surface. AB - Novel atomic structures on a H-terminated Si(100)-(2x1)-H surface were found using scanning tunneling microscopy (STM). The structures are distinguishable only from Si dimers in empty-state STM images. They were observed on arsenic- and phosphorus-doped substrates, but not on boron-doped substrates. Surface density of these structures was found to be proportional to the dopant density in the substrate. First-principles calculations clarify that they are consisting of dopant pairs that are segregated from the bulk material. Hydrogen atoms attached to the dopant pair are found to flip between two positions on the surface due to a quantum effect. PMID- 12732054 TI - Adatom transport on strained Cu(001): surface crowdions. AB - Surface strain is often suggested as a means to control the self-assembled growth of nanostructures. Strain affects both the kinetics of nucleation and the free energies of formation of the desired nanostructure. It is demonstrated here that diffusion on some strained surfaces may be mediated by newly identified adatom transport mechanism: the formation and motion of a surface crowdion. PMID- 12732055 TI - Water condensation kinetics on a hydrophobic surface. AB - Employing thermal desorption spectroscopy, we show that the effective probability of water condensation at low water vapor pressure on an octane film is much below unity at 100-120 K. This unusual finding is related to a small binding energy of H2O monomers on octane (approximately equal to 0.08 eV), requiring the formation of critical water clusters for condensation to occur. This results in strong temperature and impingement-rate dependencies of the water condensation rate and a nonlinear uptake as a function of dose time. All these features are rationalized quantitatively by a kinetic model of water condensation. PMID- 12732056 TI - Capillary force on a nanoscale tip in dip-pen nanolithography. AB - Monte Carlo simulation has been used to characterize the capillary force due to the condensation of a liquid meniscus between a tip with a nanoscale asperity and a flat surface. To consider both hydrophobic and hydrophilic molecules coating the tip as a model of dip-pen nanolithography, tips with various wettabilities are studied. The capillary force due to the meniscus is calculated for various saturations (humidities). We have implemented a thermodynamic integration technique that can project the force into energetic and entropic contributions. In most cases, the force is mainly energetic in origin. At the snap-off separation where the meniscus disappears, the tip feels a significant entropic force at high saturation. Our calculation shows nonmonotonic behavior of the pull off force as a function of saturation, which is in qualitative accord with experiments. PMID- 12732057 TI - Tuning carbon nanotube band gaps with strain. AB - We show that the band structure of a carbon nanotube (NT) can be dramatically altered by mechanical strain. We employ an atomic force microscope tip to simultaneously vary the NT strain and to electrostatically gate the tube. We show that strain can open a band gap in a metallic NT and modify the band gap in a semiconducting NT. Theoretical work predicts that band gap changes can range between +/-100 meV per 1% stretch, depending on NT chirality, and our measurements are consistent with this predicted range. PMID- 12732058 TI - Reentrant metallic behavior of graphite in the quantum limit. AB - Magnetotransport measurements performed on several well-characterized highly oriented pyrolitic graphite and single crystalline Kish graphite samples reveal a reentrant metallic behavior in the basal-plane resistance at high magnetic fields, when only the lowest Landau levels are occupied. The results suggest that the quantum Hall effect and Landau-level-quantization-induced superconducting correlations are relevant to understand the metalliclike state(s) in graphite in the quantum limit. PMID- 12732059 TI - Metal-to-semiconductor transition in squashed armchair carbon nanotubes. AB - We investigate electronic transport properties of the squashed armchair carbon nanotubes, using tight-binding molecular dynamics and the Green's function method. We demonstrate a metal-to-semiconductor transition while squashing the nanotubes and a general mechanism for such a transition. It is the distinction of the two sublattices in the nanotube that opens an energy gap near the Fermi energy. We show that the transition has to be achieved by a combined effect of breaking of mirror symmetry and bond formation between the flattened faces in the squashed nanotubes. PMID- 12732060 TI - Observation of retardation effects in the spectrum of two-dimensional plasmons. AB - Retardation effects, theoretically predicted more than 35 years ago, are observed in the spectrum of two-dimensional plasmons in high-electron-mobility GaAs/AlGaAs quantum wells. In zero magnetic field, a strong reduction of the resonant plasma frequency is observed due to the hybridization of the plasma and light modes. In a perpendicular magnetic field B, hybrid cyclotron-plasmon modes appear with a very unusual dependence of the frequency on B field. Experimental results are in excellent agreement with the theory. PMID- 12732061 TI - Measuring the transmission phase of a quantum dot in a closed interferometer. AB - The electron transmission through a closed Aharonov-Bohm mesoscopic solid-state interferometer, with a quantum dot (QD) on one of the paths, is calculated exactly for a simple model. Although the conductance is an even function of the magnetic flux (due to Onsager's relations), in many cases one can use the measured conductance to extract both the amplitude and the phase of the "intrinsic" transmission amplitude t(D)=-i|t(D)|e(ialpha(D)) through the "bare" QD. We also propose to compare this indirect measurement with the (hitherto untested) direct relation sin((2)(alpha(D)) identical with |t(D)|(2)/max((|t(D)|(2)). PMID- 12732062 TI - Superconductivity caused by the pairing of plutonium 5f Eelectrons in PuCoGa5. AB - On the basis of electronic structure calculations we identify the superconductivity in the novel, high-temperature superconductor PuCoGa5 to be caused by the pairing of Pu 5f electrons. Assuming delocalized Pu 5f states, we compute theoretical crystallographic constants very near to the experimental ones, and the calculated specific heat coefficient compares reasonably to the measured coefficient. The theoretical Fermi surface is quasi-two-dimensional and the material appears to be close to a magnetic phase instability. PMID- 12732063 TI - Experimental evidence for anisotropic double-gap behavior in MgB2. AB - The behavior of a type II superconductor in the presence of a magnetic field is governed by two characteristic length scales, the London penetration depth and the coherence length. We present magnetization measurements on MgB2 powder showing an anisotropy in the upper critical field and hence the coherence length of 6. Using the technique of small angle neutron scattering we show that this anisotropy is not mirrored in the London penetration depth, which is almost isotropic. This result can be explained by the superconductivity residing in two distinct electronic bands of the material, only one of which is highly anisotropic. PMID- 12732064 TI - Topological gauge structure and phase diagram for weakly doped antiferromagnets. AB - We show that a topological gauge structure in an effective description of the t-J model gives rise to a global phase diagram of antiferromagnetic and superconducting phases in a weakly doped regime. Dual confinement and deconfinement of holons and spinons play essential roles here, with a quantum critical point at a doping concentration x(c) approximately equal to 0.043. The complex experimental phase diagram at low doping is well described within such a framework. PMID- 12732065 TI - Field dependence of the muon spin relaxation rate in MnSi. AB - Muon spin rotation/relaxation measurements have been performed in the itinerant helical magnet MnSi at ambient pressure and at 8.3 kbar. We have found the following: (a) the spin-lattice relaxation rate 1/T(1) shows divergence as T1T proportional, variant (T-T(c))(beta) with the power beta larger than 1 near T(c); (b) 1/T(1) is strongly reduced in an applied external field B(L) and the divergent behavior near T(c) is completely suppressed at B(L)> or =4000 G. We discuss that (a) is consistent with the self-consistent renormalization theory and reflects a departure from "mean-field" behavior, while (b) indicates selective suppression of spin fluctuations of the q=0 component by B(L). PMID- 12732066 TI - Doped coupled frustrated spin-1/2 chains with four-spin exchange. AB - The role of various magnetic interchain couplings is investigated by numerical methods in doped frustrated quantum spin chains. A nonmagnetic dopant introduced in a gapped spin chain releases a free spin-1/2 soliton. The formation of a local magnetic moment is analyzed in terms of soliton confinement. A four-spin coupling which might originate from cyclic exchange is shown to produce such a confinement. Dopants on different chains experience an effective space-extended nonfrustrating pairwise spin interaction. PMID- 12732067 TI - Pseudogap Fermi-Bose Kondo model. AB - We consider a magnetic impurity coupled to both fermionic quasiparticles with a pseudogap density of states and bosonic spin fluctuations. Using renormalization group and large-N calculations we investigate the phase diagram of the resulting Fermi-Bose Kondo model. We show that the Kondo temperature is strongly reduced by low-energy spin fluctuations, and make connections to experiments in cuprate superconductors. Furthermore, we derive an exact exponent for the critical behavior of the conduction electron T matrix, and propose our findings to be relevant for certain scenarios of local quantum criticality in heavy-fermion metals. PMID- 12732068 TI - Origin of the fine structure of the Raman D band in single-wall carbon nanotubes. AB - Experiments show that the D bands of bundles of single wall carbon nanotubes have a fine structure, apparently consisting of more than one subband. Using the double resonance theory, we calculate for the first time the D band for a sample of a given diameter distribution for seven different laser excitation energies in a wide range. In addition, a detailed theoretical explanation for the fine structure of the D band is provided. The calculated results agree well with experiments and show that the main factors in determining the fine structure are an enhanced trigonal warping of the phonon dispersion, the presence of a diameter distribution in the sample, and--most importantly--the resonance from the Van Hove singularities. PMID- 12732069 TI - Electromechanical properties of metallic, quasimetallic, and semiconducting carbon nanotubes under stretching. AB - An electromechanical system is constructed to explore the electrical properties of various types of suspended single-walled carbon nanotubes under the influence of tensile stretching. Small band-gap semiconducting (or quasimetallic) nanotubes exhibit the largest resistance changes and piezoresistive gauge factors ( approximately 600 to 1000) under axial strains. Metallic nanotubes exhibit much weaker but nonzero sensitivity. Comparison between experiments and theoretical predictions and potential applications of nanotube electromechanical systems for physical sensors (e.g., strain gauges, pressure sensors, etc.) are discussed. PMID- 12732070 TI - Eavesdropping on the "ping-pong" quantum communication protocol. AB - Security of the "ping-pong" quantum communication protocol recently proposed by Bostrom and Felbinger [Phys. Rev. Lett. 89, 187902 (2002)]] is analyzed in the case of considerable quantum channel losses. The eavesdropping scheme is presented, which reveals that the ping-pong protocol is not secure for transmission efficiencies lower than 60%. Our scheme induces 50% losses, which, however, can be hidden in the channel losses if one replaces the original lossy channel with a less lossy one. Finally, a possible improvement of the ping-pong protocol security is proposed. PMID- 12732071 TI - Fiber-optics implementation of the Deutsch-Jozsa and Bernstein-Vazirani quantum algorithms with three qubits. AB - We report on a fiber-optics implementation of the Deutsch-Jozsa and Bernstein Vazirani quantum algorithms for 8-point functions. The measured visibility of the 8-path interferometer is about 97.5%. Potential applications of our setup to quantum communication or cryptographic protocols using several qubits are discussed. PMID- 12732072 TI - Symmetric extensions of quantum States and local hidden variable theories. AB - While all bipartite pure entangled states violate some Bell inequality, the relationship between entanglement and nonlocality for mixed quantum states is not well understood. We introduce a simple and efficient algorithmic approach for the problem of constructing local hidden variable theories for quantum states. The method is based on constructing a so-called symmetric quasiextension of the quantum state that gives rise to a local hidden variable model with a certain number of settings for the observers Alice and Bob. PMID- 12732073 TI - Bell inequalities with auxiliary communication. AB - What is the communication cost of simulating the correlations produced by quantum theory? We generalize Bell inequalities to the setting of local realistic theories augmented by a fixed amount of classical communication. Suppose two parties choose one of M two-outcome measurements and exchange 1 bit of information. We present the complete set of inequalities for M=2, and the complete set of inequalities for the joint correlation observable for M=3. We find that correlations produced by quantum theory satisfy both of these sets of inequalities. One bit of communication is therefore sufficient to simulate quantum correlations in both of these scenarios. PMID- 12732074 TI - Active traveling wave in the cochlea. AB - A sound stimulus entering the inner ear excites a deformation of the basilar membrane which travels along the cochlea towards the apex. It is well established that this wavelike disturbance is amplified by an active system. Recently, it has been proposed that the active system consists of a set of self-tuned critical oscillators which automatically operate at an oscillatory instability. Here, we show how the concepts of a traveling wave and of self-tuned critical oscillators can be combined to describe the nonlinear wave in the cochlea. PMID- 12732075 TI - Potts ferromagnets on coexpressed gene networks: identifying maximally stable partitions. AB - Clustering gene expression data by exploiting phase transitions in granular ferromagnets requires transforming the data to a granular substrate. We present a method using the recently introduced homogeneity order parameter Lambda [H. Agrawal, Phys. Rev. Lett. 89, 268702 (2002)]] for optimizing the parameter controlling the "granularity" and thus the stability of partitions. The model substrates obtained for gene expression data have a highly granular structure. We explore properties of phase transition in high q ferromagnetic Potts models on these substrates and show that the maximum of the width of superparamagnetic domain, corresponding to maximally stable partitions, coincides with the minimum of Lambda. PMID- 12732076 TI - Phase transitions in sexual populations subject to stabilizing selection. AB - We show that a simple model of an evolving sexual population, which dates back to some of the earliest work in theoretical population genetics, exhibits an unexpected and previously unobserved phase transition between ordered and disordered states. This behavior is not present in populations evolving asexually without recombination and is thus important in any comparison of sexual and asexual populations. In order to calculate the details of the phase transition, we use techniques from statistical physics. We introduce the correlation of the population as the order parameter of the system and use maximum entropy inference to find the state of the population at any time. PMID- 12732077 TI - Nucleation of crystalline phases of water in homogeneous and inhomogeneous environments. AB - We employ two-body and three-body bond-orientational order-parameters, in conjunction with non-Boltzmann sampling to calculate the free energy barrier to nucleation of crystalline phases of water. We find that, as the coupling between the successive peaks of the direct correlation function increases, the free energy barrier to nucleation decreases. On this basis we explain the important parameters that govern the nucleation rate involving crystalline phases of water in different homogeneous and inhomogeneous environments, giving a "unified picture" of ice nucleation in water. PMID- 12732078 TI - Phase transitions of colloidal monolayers in periodic pinning arrays. AB - We study the phase behavior of two-dimensional paramagnetic colloidal systems on square pinning arrays, the latter being created by a holographic optical tweezer technique. When the particle interaction strength is decreased, a transition from an incommensurate to a commensurate solid is observed. At even smaller pair potentials, the interstitial particles start to melt, whereas the particles at the substrate pinning sites are still localized. Our results are in good agreement with recent numerical studies on vortex melting in periodic pinning arrays. PMID- 12732079 TI - Phase transition in multiprocessor scheduling. AB - An "easy-hard" phase transition is shown to characterize the multiprocessor scheduling problem in which one has to distribute the workload on a parallel computer such as to minimize the overall run time. The transition can be analyzed in detail by mapping it on a mean-field antiferromagnetic Potts model. The static phase transition, characterized by a vanishing ground state entropy, corresponds to a transition in the performance of practical scheduling algorithms. PMID- 12732080 TI - Comment on "Experimental evidence of core modification in the near drip-line nucleus 23O". PMID- 12732082 TI - Comment on "Why is the DNA denaturation transition first order?". PMID- 12732084 TI - Preoperative evaluation of lymph node metastasis in esophageal cancer. AB - Lymph node metastasis (LMN) in esophageal cancer occurs from the superficial cancer and spreads wildly from the neck to the abdomen. Hence precise determination of LMN is essential when adequate treatments are employed. There are several reports about ultrasonic features of malignant lymph nodes (LNs), and in summary, reports of endoscopic ultrasonography (EUS) findings of malignant LNs showed they were more than 5-10 mm in diameter, with a distinct border, hypoechoic internal echo and round shape. Sensitivity, specificity and accuracies for the diagnosis of malignant LNs by EUS were 49-99%, 33-99% and 71-96%. The rates widely varied, because the accuracy of EUS's ability to determine malignancy were based on the evaluation of various echo features of LNs, and were dependent on the judgement of subjective observers. Therefore histological analysis is necessary for adequate treatments. Endoscopic ultrasonography guided fine-needle aspiration cytology (EUS-FNA) has been performed for the diagnosis of malignant LNs since 10 years. Results of those reports were sensitivity 81-97%, specificity 83-100% and accuracy 83-97%. EUS-FNA staging was better than EUS staging. Also clinically obvious complications by EUS-FNA have not been reported. Therefore published evidence showed that EUS-FNA is safe and useful for confirmation of malignant LNs. PMID- 12732085 TI - Changes in respiratory condition after thymectomy for patients with myasthenia gravis. AB - BACKGROUND: Anesthesia without muscle relaxants has been reported to be effective for early extubation after thymectomy, but postoperative respiratory status of the patients has not been studied intensively. METHODS: Fifty-two consecutive patients undergoing thymectomies for myasthenia gravis (MG) were evaluated. RESULTS: Forty-two (81%) of the 52 patients were extubated in the operating room, and 49 (94%) patients were extubated within 24 hours. However, 6 (12%) patients required subsequent reintubation for respiratory support. There was a sudden increase in the respiratory rate (RR) and PaCO(2). The mean value of the increase in PaCO(2) at the time of reintubation was 23 mmHg (12-58 mmHg). The mean value of the increase in RR above the preoperative level at the time of reintubation was 16/min (7-30/min). In univariate analysis, vital capacity (VC), %VC, the preoperative pyridostigmine dose and the duration of surgery correlated with reintubation, but with multivariate analyses, the pyridostigmine dose was the only significant factor related to reintubation. CONCLUSION: The patients who received at least or more than 240 mg of pyridostigmine should be monitored carefully after tracheal extubation. PMID- 12732086 TI - Prognostic significance of serum squamous cell carcinoma antigen in surgically treated lung cancer. AB - The serum concentrations of squamous cell carcinoma antigen (SCC-Ag) obtained from 124 surgically treated primary non-small cell lung cancer patients, including 75 adenocarcinomas (AD) and 49 squamous cell carcinomas (SQ), were studied. The changes in the SCC-Ag concentration, which were obtained before and one month after surgery, were analyzed. The 5-year survival rate of the patients with AD who were positive for SCC-Ag preoperatively (32%) was lower than that for those who were negative for SCC-Ag preoperatively (57%, p<0.05). Meanwhile, in those with SQ, the 5-year survival rate of those who were positive for SCC-Ag preoperatively (59%) was not different when compared with those who were negative for SCC-Ag preoperatively (73%). The 5-year survival rate of patients with AD who were positive for SCC-Ag preoperatively and negative postoperatively was 53% versus 17% for those who remained positive postoperatively (p<0.05). In those with SQ, the 5-year survival rate of those who were positive for SCC-Ag preoperatively and negative postoperatively was 76% while it was 0% for those who remained positive postoperatively (p<0.01). In patients with negative SCC-Ag postoperatively, 5-year survival rates were not different between the patients who had positive antigen preoperatively and the patients who had negative antigen preoperatively both in AD (53% and 57%, respectively) and SQ (76% and 75%, respectively). In conclusion, though SCC-Ag is widely used for SQ, preoperative SCC-Ag did not reflect the prognosis. In AD, the survival rate was lower in antigen-positive than antigen-negative patients. Survival rate was higher in antigen-positive patients who became antigen-negative following resection than in patients who remained antigen-positive for both AD and SQ. In the patients who were negative for SCC-Ag postoperatively, survival was the same regardless of the preoperative SCC-Ag positivity in both AD and SQ. PMID- 12732087 TI - Clinical effects of percutaneous cardiopulmonary support in severe heart failure: early results and analysis of complications. AB - Between January 1993 and December 2001, we employed percutaneous cardiopulmonary support (PCPS) in 35 patients. PCPS was used for postcardiotomy in 25 of these patients who could not be weaned from cardiopulmonary bypass (CPB) because of severe cardiogenic shock. In the other 10 patients, PCPS was used for a non surgical disease. Twenty-nine patients (82.9%) were weaned from PCPS, and 28 (80.0%) survived. The other 7 patients (20.0%) died due to postoperative complications. The causes of death were multiple organ failure (MOF) due to wound bleeding, low cardiac output syndrome (LOS), myonephropathic metabolic syndrome (MNMS) with severe lower limbs ischemia, cerebrovascular accident (CVA), and sepsis. The first cause for the complications was postoperative sustained severe heart failure. To improve the survival rate, it was necessary to prevent bleeding and begin PCPS at an earlier stage. PMID- 12732088 TI - Complete revascularization in patients with severe left ventricular dysfunction. AB - OBJECTIVE: This study evaluates whether patients with coronary artery disease and severely depressed left ventricular ejection fraction (LVEF) benefit from complete revascularization by multivessel coronary artery bypass. METHODS: From April 1994 to May 2002, 42 patients who underwent coronary artery bypass grafting (CABG) at our institution had impaired left ventricular (LV) function [an ejection fraction (EF) of 30% or less]. The average preoperative LVEF was 23.8%. The mean number of grafts was 4.6. Complete revascularization by multivessel bypass grafting was the goal for all patients. RESULTS: Thirty days mortality was 0 and hospital mortality was 2.4%. The mean graft patency rate for 35 (83%) patients at one month was 98.8%. The mean postoperative LVEF improved significantly, from 23.8% to 35.2% (p<0.05), and the New York Heart Association (NYHA) classification was improved in most patients. The Kaplan-Meier estimate of survival at 5 years was 83.1%, and that of the cardiac event-free rate at 5 years was 77.5%. CONCLUSION: For patients with poor LV function, complete surgical revascularization by multivessel bypass grafting can be performed safely, with satisfactory hospital mortality and long-term results. PMID- 12732089 TI - Efficacy and adverse effects of the coronary active perfusion system--from a viewpoint of perfusional timing. AB - We developed a coronary active perfusion system (CAPS) to avoid myocardial ischemia during off-pump coronary artery bypass (OPCAB). The purpose of this study was to determine the optimal timing of CAPS perfusion, and to investigate any adverse effects when it is driven within the systolic phase. Twenty-four pigs were divided into four groups: (A) systolic, (B) early-diastolic, (C) middle diastolic, and (D) late-diastolic perfusion. Myocardial blood flow (MBF), hemodynamic and mechanical data were measured during 30 minutes of CAPS perfusion. MBF in group A was lower than in the other groups (p<0.001), but no significant differences were observed among groups B to D. End-systolic pressure volume relation (Mw) and preload recruitable stroke work relation (Ees) in group A was lower than in groups B to D (p<0.01 and p<0.05, respectively), but no significant differences were observed among groups B to D. CAPS could maintain regional MBF and left ventricular function if it was driven only within the diastolic phase and required no strict adjustment, but CAPS has an adverse effect when it is driven in the systolic phase. PMID- 12732090 TI - Pulmonary granuloma possibly caused by staples after video-assisted thoracoscopic surgery. AB - A 74-year-old man, with a history of colon cancer resection and video-assisted thoracoscopic surgery (VATS) resection of pulmonary metastases, was found to have a left pulmonary nodule near the previous staple-line. The size of this nodule increased during follow-up. Because of this clinical course, this nodule was considered to be a tumor recurrence at the staple-line, and pulmonary re resection was performed. The pathologic diagnosis of this nodule was foreign body granuloma (FBG) possibly due to previous surgical staples. FBG induced by staples may be a rare complication in VATS. PMID- 12732092 TI - Surgical repair of a common atrium in an adult. AB - We report a rare successful surgical repair of a common atrium (CA) with mild tricuspid valve (TV) regurgitation due to valvular annulus enlargement in a 39 year-old man, who had a complete atrial septum defect (ASD) without the characteristic of an endocardial cushion defect. The left-to-right shunt ratio was 85 percent and the Qp/Qs was 6.7 due to the CA. Left ventriculogram revealed no evidence of typical goose-neck deformity and no mitral valve regurgitation. The operation consisted of making a new atrial septum with an autologous pericardial patch and tricuspid annuloplasty (DeVega) using extracorporeal circulation. There was no evidence of a cleft on the anterior leaflet of the mitral valve or the septal leaflet of the TV. The postoperative echocardiogram showed no residual shunt flow through a new atrial septum and no TV regurgitation, and atrioventricular (AV) dissociation did not occur. We consider this procedure to be widely applicable in consideration of the favorable results obtained after surgical treatment. PMID- 12732091 TI - Pleomorphic carcinoma: report of a case with massive pleural effusion and asbestos particles. AB - Pleomorphic (spindle/giant cell) carcinoma (PC) is one subset of large cell carcinoma. It is well known that PC patients have a poor survival rate. This report describes a 68-year-old man with PC. The patient's tumor had a massive pleural effusion. A left lower lobectomy and partial resection of the left diaphragm, peritoneum, and parietal pleura were performed to remove the tumor. Numerous asbestos particles were found in the left lower lobe. This is the first reported case of PC which may have been caused by asbestos particles. Further investigation is needed into whether asbestos exposure causes PC. PMID- 12732093 TI - Congenital quadricuspid aortic valve: report of nine surgical cases. AB - We report the clinicopathologic characteristics of the congenital quadricuspid aortic valve necessitating surgery. Among 616 patients for whom we performed an aortic valve operation over the past 20 years, nine patients (1.46%) (five men and four women, mean age 60 years) with quadricuspid aortic valve were encountered. All had aortic regurgitation (AR) except one with aortic stenosis and mild regurgitation (ASr). All were free of cardiac anomaly including that of the coronary arterial system. Macroscopically, severe calcification of the valve was seen in the one case of ASr. Fenestration of the cusp was seen in five cases of AR. Infective endocarditis was not seen. Histological study disclosed fibrous thickening and myxoid degeneration in the AR cases. In accordance with the Hurwitz and Roberts classification, four valves were type b (three equal-sized cusps and one smaller cusp), two valves were type a (four equal-sized cusps), two valves were type d (one large, two intermediate, and one small cusp), and one valve was type g (four unequal-sized cusps). Valve repair failed in one patient and was converted to valve replacement during the operation. All patients underwent successful aortic valve replacement (AVR). PMID- 12732094 TI - Batista operation with aortic valve replacement for valvular cardiomyopathy. AB - We performed a successful Batista operation with aortic valve replacement (AVR), graft replacement of the ascending aorta, and tricuspid annuloplasty for a patient with valvular cardiomyopathy with severe aortic stenosis, an ascending aortic aneurysm, and grade 3 tricuspid regurgitation. PMID- 12732095 TI - A new probe for the anastomosis of small vessels. AB - We developed new probes with three varying sizes (phi1.0, 1.5 and 2.0 mm) with a trench for guiding the needle which made small vessel anastomoses easy and suture placement accurate with clear identification of the vessel lumen. We evaluated the efficiency of these probes on anastomoses using the bilateral common carotid arteries of cadaver rabbits. The anastomosis time of end-to-side anastomosis was shortened by using the probe from 20.2+/-3.3 to 15.4+/-2.6 min, and of side-to side anastomosis from 20.2+/-1.3 to 16.0+/-1.2 min. Of the 5 end-to-side anastomoses without the probe, there was one deformity of the anastomoses site and of the 5 side-to-side anastomoses without the probe, there was one stenosis of the anastomosis. There were neither deformity nor stenosis of the 10 anastomoses of 5 end-to-side and 5 side-to-side anastomoses with the probes. In conclusion, the probe with a trench for guiding the needle made small vessel anastomoses easy. PMID- 12732096 TI - Characterization of the carbohydrate moieties of the functional unit RvH1-a of Rapana venosa haemocyanin using HPLC/electrospray ionization MS and glycosidase digestion. AB - The primary structures of two biantennary N -glycans of the glycoprotein Rapana venosa (marine snail) haemocyanin were determined. Two different structural subunits have been found in R. venosa haemocyanin: RvH1 and RvH2. The carbohydrate content of the N-terminal functional unit RvH1-a of RvH1 was studied and compared with the N-terminal functional unit RvH2-a of RvH2. Oligosaccharide fragments were released from the glycoprotein by Smith degradation of a haemocyanin pronase digest and separated on a Superdex 300 column. The glycopeptide fragments, giving a positive reaction for the orcinol/H2SO4 method, were separated by HPLC. In order to determine the linked sugar chains to the hinge glycopeptides isolated from the functional unit RvH1-a, several techniques were applied, including capillary electrophoresis, matrix-assisted laser desorption ionization-MS and electrospray ionization-MS in combination with glycosidase digestion. On the basis of these results and amino acid sequence analysis, we concluded that the functional unit RvH1-a contains 7% oligosaccharides N-glycosidically attached to Asn262 and Asn401, and the following structures were suggested:[structure: see text] PMID- 12732099 TI - [Costs of digestive endoscopy in a level II university hospital]. AB - OBJECTIVES: To establish the criteria that should be considered when analyzing the cost of digestive endoscopy and to determine how the variables studied influence the final results, as well as to determine the relative value unit (RVU) per endoscopic procedure. MATERIAL AND METHOD: Clinical management study relating the cost of endoscopic procedures with their complexity, healthcare activity and direct and indirect countable costs. The endoscopic procedures performed from 2000-2001 (4,982 procedures) were analyzed. We determined the staff costs according to the hours devoted to endoscopic activity, the procedures performed and their complexity, non-amortizable and amortizable materials acquired in the study period, and the cost and amortization of apparatus and equipment. RESULTS: The biannual cost was 392,892.60;. Staff costs were 63%, apparatus and equipment 15%, structural costs 13%, pharmacy 6%, materials 2% and amortizable materials 1%. The least expensive procedure was diagnostic gastroscopy (60.56;) and the most expensive was therapeutic endoscopic retrograde cholangiopancreatography (277.06;). The RVU cost was 52.58;. CONCLUSIONS: Calculation of the cost of any medical procedure should take into account the strict application of direct and indirect costs. In our environment, the cost of endoscopy is lower than might be expected, mainly because the cost of amortization of apparatus and equipment and staff costs were low. Calculation of the complexity index is of considerable clinical and healthcare value. Determination of the RVU is a key element in establishing the cost of a procedure and in relating this cost with other costs, allowing its application as well as comparison among different investigations, services and centers. PMID- 12732098 TI - Wavelet to predict bacterial ori and ter: a tendency towards a physical balance. AB - BACKGROUND: Chromosomal DNA replication in bacteria starts at the origin (ori) and the two replicores propagate in opposite directions up to the terminus (ter) region. We hypothesize that the two replicores need to reach ter at the same time to maintain a physical balance; DNA insertion would disrupt such a balance, requiring chromosomal rearrangements to restore the balance. To test this hypothesis, we needed to demonstrate that ori and ter are in a physical balance in bacterial chromosomes. Using wavelet analysis, we documented GC skew, AT skew, purine excess and keto excess on the published bacterial genomic sequences to locate the turning (minimum and maximum) points on the curves. Previously, the minimum point had been supposed to correlate with ori and the maximum to correlate with ter. RESULTS: We observed a strong tendency of the bacterial chromosomes towards a physical balance, with the minima and maxima corresponding to the known or putative ori and ter and being about half chromosome separated in most of the bacteria studied. A nonparametric method based on wavelet transformation was employed to perform significance tests for the predicted loci. CONCLUSIONS: The wavelet approach can reliably predict the ori and ter regions and the bacterial chromosomes have a strong tendency towards a physical balance between ori and ter. PMID- 12732097 TI - Human aldose reductase and human small intestine aldose reductase are efficient retinal reductases: consequences for retinoid metabolism. AB - Aldo-keto reductases (AKRs) are NAD(P)H-dependent oxidoreductases that catalyse the reduction of a variety of carbonyl compounds, such as carbohydrates, aliphatic and aromatic aldehydes and steroids. We have studied the retinal reductase activity of human aldose reductase (AR), human small-intestine (HSI) AR and pig aldehyde reductase. Human AR and HSI AR were very efficient in the reduction of all- trans -, 9- cis - and 13- cis -retinal ( k (cat)/ K (m)=1100 10300 mM(-1).min(-1)), constituting the first cytosolic NADP(H)-dependent retinal reductases described in humans. Aldehyde reductase showed no activity with these retinal isomers. Glucose was a poor inhibitor ( K (i)=80 mM) of retinal reductase activity of human AR, whereas tolrestat, a classical AKR inhibitor used pharmacologically to treat diabetes, inhibited retinal reduction by human AR and HSI AR. All- trans -retinoic acid failed to inhibit both enzymes. In this paper we present the AKRs as an emergent superfamily of retinal-active enzymes, putatively involved in the regulation of retinoid biological activity through the assimilation of retinoids from beta-carotene and the control of retinal bioavailability. PMID- 12732101 TI - [Presinusoidal portal hypertension due to portal thrombosis in a patient with Alagille's syndrome]. AB - We present the case of a 16-year old woman with Alagille's syndrome, who had upper gastrointestinal bleeding due to rupture of esophageal varices secondary to presinusoidal portal hypertension without liver fibrosis. Portal thrombosis is a manifestation previously unreported in association to this syndrome. PMID- 12732100 TI - [Value of C-reactive protein level in the detection of necrosis in acute pancreatitis]. AB - OBJECTIVES: To evaluate the utility of C-reactive protein (CRP) in the detection of necrosis in acute pancreatitis and to determine the best cut-off point for CRP used for this purpose. MATERIAL AND METHODS: We performed a retrospective study of 157 patients with acute pancreatitis who underwent computed tomography (CT) with intravenous contrast material between 72 h and 8 days after the onset of symptoms and whose serum CRP values were determined by nephelometry 24 h or later after the onset of pain. RESULTS: Ninety-four patients were men and 63 were women, with a mean age of 61 years (range, 15-96 years). The cause of pancreatitis was biliary lithiasis in 53.5%, alcohol in 20.4%, and idiopathic in 10.8%. Other causes were found in 15.3%. The mean (standard deviation) time elapsed between symptom onset and extraction to evaluate CRP was 3.21 (1.7) days. The patients were divided into two groups according to the results of CT: 132 patients with acute intersitial edematous pancreatitis and 25 patients with acute necrotizing pancreatitis. The mean CRP concentrations were: 322 mg/l (range, 10.7 538) in patients with acute necrotizing pancreatitis and 133 mg/l (range, 3-442) in those with acute interstitial pancreatitis; this difference was statistically significant (p < 0.001). The area under the ROC curve of CRP vs. the occurrence of necrosis was 0.862 (95% CI, 0.778, 0.946). To evaluate the presence of pancreatic necrosis a cut-off level of 200 mg/l showed a sensitivity of 88% and a specificity of 75% while a cut-off level of 279 mg/l presented a sensitivity of 72% and a specificity of 88%. CONCLUSIONS: In acute pancreatitis, a CRP value of less than or equal to 200 mg/l obtained at 72 h of symptom onset is useful for ruling out, with a high degree of probability, the presence of necrosis. With higher values, additional investigations should be performed to determine the presence of pancreatic necrosis; nevertheless, with values higher than 279 mg/l the risk of necrosis markedly increases. PMID- 12732102 TI - [Ascites as the sole clinical manifestation in a patient with nodular regenerative hyperplasia]. AB - Nodular regenerative hyperplasia (NRH) of the liver is an infrequent entity that is usually diagnosed after the appearance of clinical signs of portal hypertension such as hepatomegaly, splenomegaly or upper gastrointestinal bleeding due to esophageal varices, which are the most frequently found clinical manifestations in NRH. Ascites is a less frequent finding and has always been described in association with other manifestations of portal hypertension. We describe a new case of NRH with atypical presentation in which ascites was the sole clinical manifestation. PMID- 12732103 TI - [Chronic pancreatitis as the initial presentation of Crohn's disease]. AB - It has been suggested that pancreatitis could be an extrahepatic manifestation of inflammatory bowel disease, since its incidence in this disease is greater than that in the general population and in many cases no etiological factor is found. We present a case of chronic idiopathic pancreatitis as the initial presentation of Crohn's disease of the colon. PMID- 12732104 TI - [Antiviral treatment in liver cirrhosis due to hepatitis C virus]. PMID- 12732105 TI - [Genetic polymorphisms and liver diseases]. PMID- 12732107 TI - [The role of anti-neutrophil cytoplasmic antibodies (ANCA) and anti-Saccharomyces cerevisiae antibodies (ASCA) in inflammatory bowel disease]. PMID- 12732106 TI - [The 65th birthday of Juan Rodes. Speech delivered in honor of Juan Rodes, celebrated at the Congress of the Spanish Society for Liver Studies, Barcelona, 28 February 2003]. PMID- 12732109 TI - [Peripapillary lipomas "in cluster"]. PMID- 12732108 TI - [Helicobacter pylori eradication. Triple or quadruple therapy?, Long or short treatment?]. PMID- 12732110 TI - [Acinetobacter baumannii: a nosocomial pathogen difficult to control]. PMID- 12732111 TI - [Incidence, risk factors and influence on survival of infectious complications in liver transplantation]. AB - INTRODUCTION: Orthotopic liver transplantation (OLT) is successful therapy for patients with end-stage liver disease. Infection is currently a life-threatening complication for these patients. The aims of this study are to determine the incidence of various infections in patients with OLT, to study overall survival rates and survival as related to individual infections, and to investigate the risk factors associated with first episodes of bacterial (BI), fungal (FI), invasive fungal (IFI) and cytomegalovirus (CMV) infections. METHODS: The study includes 165 OLTs performed in 152 recipients from May 1994 to May 1998. A descriptive analysis estimating the 95% confidence interval was performed with 100 variables stratified according to preoperative, operative and postoperative conditions. Cox regression analysis was used to identify the variables associated with infection. Survival studies were carried out with the Kaplan-Meier method. RESULTS: Among the total, 66% of patients developed infection: 41.8% viral, 33.9% BI, 20.6% FI and 4.2% IFI. One-year and 4-year survival rates after transplantation were 90% and 75%, respectively. All the infections decreased survival. Multivariate analyses identified the following risk factors for the specific infections: BI - dialysis, mechanical ventilation, and time of organ ischemia during harvesting; FI - number of hours of surgery and pretransplantation plasma albumin concentrations; IFI - number of blood units transfused, pretransplantation plasma albumin and retransplantation. Cytomegalovirus infection was associated with FI and IFI in the univariate analysis, but the multivariate analysis identified no variables that independently increased the risk of developing this infection. PMID- 12732112 TI - [Orthopedic implant infection: prognostic factors and influence of long-term antibiotic treatment on evolution. Prospective study, 1992-1999]. AB - INTRODUCTION: Orthopedic implant infections are significant because of their morbidity, a tendency to serious relapses and an elevated health cost. OBJECTIVES: To study prognostic factors and the influence of long-term antibiotic treatment on the evolution of orthopedic implant infections. METHODS: This prospective study was performed in 110 patients with orthopedic implant infections. Clinical, analytical, and microbiological studies, as well as gammagraphy with Tc, Ga and labeled leukocytes, were performed in all patients. Controls were carried out at 7, 15 and 30 days after starting treatment, every 3 months thereafter until the end of treatment, and every 6 months thereafter up to one year after stopping treatment. Initial antibiotic treatment was prescribed according to the epidemiological characteristic of the type of infection and was modified according to the microorganism isolated. Duration of treatment was established by patient and implant characteristics, severity of infection and evolution of the process, and it was adjusted to criteria of cure, failure and relapse. RESULTS: Among the 110 cases, 63 were women and 37 men, with a mean age of 59.6 years (range 18-79 years). Implants included 72 joint prostheses (42 knee, 29 hip and 1 shoulder) and 38 bone implants. Microbiological documentation was obtained in 60%; among these, 60.6% were gram-positive cocci, with a predominance of staphylococci, 33.3% were gram-negative bacilli and 6.1% were anaerobic microorganims. Prognostic factors significantly associated with failure or relapse included previous joint surgery, previous hospital stay longer than 15 days, diabetes, microbiological isolation and treatment with cefuroxim plus rifampicin. Mean treatment duration was 9.8 months (range 2-17 months). Antibiotic treatment consisted of the following: 61 cases received fluorquinolones or cotrimoxazole plus rifampicin, 29 received cefuroxime-axetil plus rifampicin and the remaining 20 received monotherapy. Among 110 cases, 91 cured (83%). Treatment failures or relapses were observed in 19 (17%) patients; 26.7% of the latter were related to the implants. Tolerance to long-term antibiotic treatment was good.Conclusion. Long-term antibiotic treatment lasting a mean of 9.8 months had a positive influence on the evolution of orthopedic implant infections. PMID- 12732113 TI - [Nosocomial diarrhea outbreak due to Clostridium difficile in a vascular surgery department]. AB - INTRODUCTION: Clostridium difficile is considered the most common cause of nosocomial-acquired diarrhea. In Spain this condition is rarely reported. METHODS: Over a five-month period, 35 episodes of C. difficile diarrhea were diagnosed by toxin A detection in stool samples. Case-control studies were designed to assess risk factors for the outbreak. Twelve cases from the vascular surgery department were compared with 24 randomized controls, patients admitted to the same ward during this period, but without diarrhea. Statistical comparisons (SPSS software) were performed with the Student's t and X2 tests (OR with 95% CI). RESULTS: Overall incidence was 3.42 episodes: 48 episodes/1000 admissions to vascular surgery. Among the total, 80% had received antibiotics before admission and the mean number of antibiotics administered was 2.91 per patient. Comparative analyses disclosed that the cases had received a larger number of antibiotics during hospitalization (p 5 0.000) and in the two months before admission (p 5 0.031) than the controls. Clindamycin administered alone (p 5 0.001) or associated with aztreonam (p 0.000) were the most frequently used antibiotic treatments. CONCLUSIONS: C. difficile diarrhea is common in our setting. Diagnosis requires a high index of suspicion. We attribute the nosocomial outbreak in our vascular surgery department to broad-spectrum antibiotic use (clindamycin and aztreonam). Surveillance, together with restriction of antibiotic use was effective for control. PMID- 12732114 TI - [Acinetobacter baumannii bacteremia: clinical and prognostic features]. AB - OBJECTIVE: The incidence, clinical features, and prognosis of bacteremia due to Acinetobacter baumannii were investigated. METHODS: Prospective study of all episodes of A. baumanii bacteremia detected during the period of January 1995 to December 1998. A. baumannii was identified using recent standard methods. RESULTS: A total of 133 episodes of bacteremia due to A. baumannii were studied, all of them nosocomial-acquired. The incidence-density diminished from 2.02 episodes per 10,000 patient-days to 0.40 episodes per 10,000 patient-days after the implementation of a control program. Most of the patients (70%) were, or had been, in the ICU when bacteremia occurred. Some 80% of patients had a chronic illness and 62% had a Hilf's severity score > 4. Among the strains identified, 74% were multidrug-resistant and 28% were imipenem-resistant. Attributable mortality was 25.6%. Multivariate analysis showed that inappropriate antibiotic treatment, septic shock, and high Hilf's severity score were associated with poorer prognosis. CONCLUSION: A. baumannii bacteremia mainly affects severely ill patients who have undergone several invasive procedures, and who may have relevant associated morbidity and mortality. Among other variables, inappropriate antibiotic treatment was a risk factor for increased mortalilty. PMID- 12732116 TI - [Quinolones]. AB - Quinolones act by inhibiting enzymes (topoisomerases), which are indispensable to DNA synthesis. Their bactericidal activity is concentration-dependent. Their spectrum has become broader, especially since the introduction of a fluorine atom at position 6 (fluoroquinolones). They are used as the treatment of choice or as alternative therapy in a wide variety of infections, both in the hospital and non hospital setting. Depending on the compound, they are used in urinary tract infections, sexually transmitted diseases, chronic osteomyelitis, respiratory tract infections, and severe systemic infections, among others. The upsurge and extent of quinolone resistance has limited the use of these agents in some cases and in future may determine their use in others. There are strategies to minimize the spread of resistance. Quinolones are safe and well tolerated. The most frequent adverse effects are gastrointestinal and those affecting the central nervous system. PMID- 12732117 TI - [Vesicular/pustular lesions in a patient with atopic dermatitis]. PMID- 12732115 TI - [Malaria prevention in international travel]. AB - For travelers malaria represents the principal infectious risk of severe complications and death. Infection during traveling depends on the geographical area visited, the predominant species of parasite, the frequency of resistance to antimalarial agents, and whether preventive measures have been taken. Until a vaccine has been developed, prevention strategies consist of providing travelers with information, the use of barrier methods against vector bites, the correct use of chemoprophylaxis, and the possibility of self-diagnosis and treatment. The choice of chemoprophylaxis regimen should be individualized since no regimen guarantees 100% protection or is free of adverse effects or contraindications. The most effective drugs are doxycycline, atovaquone-proguanil and mefloquine while those producing severe adverse effects with the least frequency are atovaquone-proguanil and doxycycline. PMID- 12732118 TI - [Nine-year-old child with chronic abdominal pain]. PMID- 12732119 TI - [Laryngeal histoplasmosis as a manifestation of AIDS]. PMID- 12732120 TI - [Mycobacterium lentiflavum infection: case history and review of the medical literature]. PMID- 12732121 TI - [Endocarditis due to Enterococcus casseliflavus]. PMID- 12732122 TI - [Atypical radiologic pattern in Pneumocystis carinii pneumonia]. PMID- 12732123 TI - [Pneumonia due to Neisseria meningitidis serogroup Y]. PMID- 12732124 TI - [Clinical prediction of endoscopic signs in active or recent upper gastrointestinal bleeding]. AB - BACKGROUND AND OBJECTIVE: The aim of this study was to construct and validate a mathematical model, based on clinical and laboratory data, that could be useful in the emergency department (ED) to predict which patients with upper gastrointestinal bleeding (UGB) have an active or recent bleeding. PATIENTS AND METHOD: During a period of 12 months, we included all consecutive cases of UGB that came to the ED of an urban hospital. These patients made up the primary or model obtaining series. During the 12 following months, we selected a sample of UGB patients who made up the secondary series. The mathematical predictive model was built using logistic regression analysis. RESULTS: 623 patients made up the primary series and 251 the secondary series. A score equal or higher than 4 indicated the best diagnosis accuracy (63.6%) with positive and negative predictive values of 65.1 and 40.0%, respectively. The discriminative power of the model was significant (p < 0.001) but it displayed little accuracy. CONCLUSIONS: Shock, positive tilt test, increased uremia, previous hematemesis, to be an usual alcohol drinker, presence of sweating, and no previous treatment with antiulcer drugs were independent predictors for active or recent bleeding. The diagnostic accuracy of the model does not not allow its systematic use in clinical practice. However, it may be helpful for the triage of upper gastrointestinal bleeding in the ED. PMID- 12732125 TI - [Prevalence of obesity in Spain: results of the SEEDO 2000 study]. AB - BACKGROUND AND OBJECTIVE: Obesity is a major public health problem in developed countries. It is of key importance to ascertain its magnitude and to identify main groups at risk. In this paper the prevalence of obesity in Spanish adult population aged 25-60 is estimated, based on measured height and weight. SUBJECTS AND METHOD: Pooled-analysis of regional cross-sectional nutrition surveys performed on representative random population samples. Randomly selected 9,885 free living subjects aged 25-60 years respondents of population nutritional surveys carried out in eight Spanish regions (Andalucia, Balearic Islands, Basque Country, Canary Islands, Catalunya, Galicia, Madrid and Valencia) between 1990 to 2000. Weight and height were measured on each individual by trained observers following standardised procedures and measuring instruments. The samples were pooled together and weighted according to the distribution of Spanish adult population aged 25-60 years. Obesity was defined as body mass index (BMI) >= 30 kg/m2. The protocol used in each survey was in accordance to the recommendations of the Spanish Society for the study of Obesity (SEEDO) to estimate the prevalence of obesity in population studies. RESULTS: The prevalence of obesity in Spanish adult population was 14.5% (95% CI, 13.93-15.07%), significantly higher among women 15.75% (95% CI, 14.89-16.61%), than men 13,39% (95% CI, 11.84 14.94%) (*2 = 12.470; p = 0.000). Prevalence of obesity significantly increased with age in men and women. The highest rates were estimated for the age group older than 55 years, both among males and females, 21.58% (95% CI, 18.68-24.48%) and 33.9% (95% CI, 32.73-35.07%), respectively. CONCLUSION: Obesity is a health problem which affects an important proportion of the Spanish adult population. Considering its potential impact on Public Health, it would be required to design and implement effective strategies aimed at the early detection of subjects at risk and the provision of adequate treatment, as well as to establish suitable preventive programmes. PMID- 12732127 TI - [Severe Acute Respiratory Syndrome. A new emergent infectious disease]. PMID- 12732126 TI - [Prospective multicenter study of adverse drug reactions in hospitalized elderly patients]. AB - BACKGROUND AND OBJECTIVE: The incidence of adverse drug reactions (ADRs) in hospitalized elderly patients is an important clinical problem. The purpose of this study was to determine the incidence of ADRs in elderly in-patients, to analyze the factors involved in their presentation and to evaluate the reactions detected. PATIENTS AND METHOD: Prospective multicenter study in patients older than 65 years during their stay in hospital or nursing home. The assessment consisted of a complete geriatric evaluation and a protocol for collecting information on suspected ADRs during hospitalization or nursing home stay. A multivariate analysis by multiple logistic regression was performed. RESULTS: The study included 865 patients: 185 (21%) from 5 hospital units; 325 (38%) from 8 convalescent centers; and 355 (41%) from 8 long-term centers. The incidence of ADRs was 9%. In acute units the incidence was 15%; in post-acute units, 5%; and in long-term centers, 10% (p < 0.004). In the global multivariate analysis, the risk of experiencing an ADR was associated with a greater use of medications (OR = 1.15; 1.07 1.23 for each additional drug), the presentation of delirium (OR = 3.6; 1.95-6.85) and the type of unit (acute OR = 2.6; 1.16-6.01; long-term OR = 3.3; 1.62-7.05). Fifty type A ADRs were detected (65%). With regard to severity, 36 (47%) were moderate, 27 (35%) were mild, and 14 (18%) were severe. Causality: 58 (76%) probable and 17 (20%) possible. Potential avoidability: 39 (51%) were unavoidable and 38 (49%) were totally or partially avoidable; of these, 15 (39%) were the result of an interaction with another drug. CONCLUSIONS: The incidence of ADRs in our study was close to 10%. ADRs were associated with the frequency of use, presence of delirium and type of unit, and occurred most frequently in acute and long-term units. ADRs were principally type A, moderate severity, probable causality and partially avoidable. PMID- 12732129 TI - [Severe acute respiratory syndrome]. PMID- 12732128 TI - [Autoimmune lymphoproliferative syndrome: molecular diagnosis in two families]. AB - BACKGROUND AND OBJECTIVE: The autoimmune lymphoproliferative syndrome (ALPS) is a disorder caused by a defect in lymphocytes' apoptosis and characterized by non malignant lymphoproliferation, autoimmune features and increased TCR alpha + CD4CD8 cells. Most patients have a mutation in the TNFRSF6 gene, which encodes the Fas protein. Our aim was to identify mutations in this gene in two families with possible ALPS cases. PATIENTS AND METHOD: Two patients with suspicion of ALPS, belonging to two unrelated families, were studied. To confirm such a diagnosis, immunoglobulin quantification, cellular phenotypic analysis by flow cytometry, IL-10 quantification, an apoptosis study, and molecular analysis were performed. RESULTS: Both patients showed hypergammaglobulinemia and an increased percentage of TCR alpha + CD4CD8 cells (family A patient: 14%; family B patient: 4.25%). In family A, in vitro Fas-mediated apoptosis was absent in the patient and markedly reduced in his father. In this family, both the patient and his father were heterozygous for the Fas mutation T1045C (Leu 268 Pro). The family B patient and her mother showed the Fas mutation G943T (Arg 234 Leu), both being heterozygous for it too. Both mutations are located in exon 9 of TNFRSF6 gene, affecting the death domain of the Fas protein. CONCLUSIONS: The molecular study of these families confirms a diagnosis of ALPS and suggests that the causing defect of this syndrome is compatible with an autosomal dominant inheritance with incomplete penetrance. PMID- 12732130 TI - [Impact of hormonal deficit and cardiovascular risk factors on life expectancy in hypopituitarism]. PMID- 12732131 TI - [Malaria in children and other infectious agents]. PMID- 12732132 TI - [Cyst of the thyroid cartilage]. PMID- 12732133 TI - [Abuse potential of sibutramine]. PMID- 12732134 TI - p53 and TGF-beta in development: prelude to tumor suppression? AB - Recent work in Xenopus embryos reveals an unexpected developmental role for the tumor suppressor gene p53. This finding may have implications for the evolution of p53, its interaction with Smads in TGF-beta dependent mesoderm specification, and the cooperation among p53 family members. PMID- 12732135 TI - tRNA structure goes from L to lambda. AB - In this issue of Cell, Ishitani et al. (2003) report, in a crystal, a new L-like structure of tRNA designated as lambda-form, where disruption of universal tertiary interactions is compensated by interactions with an enzyme that makes a base modification at the corner of the L. PMID- 12732137 TI - The stem cell concept in plants: a matter of debate. AB - Throughout their life, which can last for over a thousand years, plants have the fascinating ability to give rise to new organs from founder cells in their apical meristems. Whether these founder cells are equivalent to the pluripotent stem cells in animals has been a long-standing controversy amongst plant scientists. Here, this controversy will be addressed in light of classical observations and recent findings. PMID- 12732136 TI - Coordinate regulation of an extended chromosome domain. AB - Spitz et al (2003[this issue of Cell]) describe the properties of a novel cis regulatory DNA element, the global control region (GCR), which regulates gene expression over distances of several hundred kilobases at the mouse HoxD complex. The GCR provides an explanation for the colinear genetic linkage and expression of individual Hox genes within developing limbs. PMID- 12732138 TI - Stereotyped pruning of long hippocampal axon branches triggered by retraction inducers of the semaphorin family. AB - Like naturally occurring neuronal cell death, stereotyped pruning of long axon branches to temporary targets is a widespread regressive phenomenon in the developing mammalian brain that helps sculpt the pattern of neuronal connections. The mechanisms controlling stereotyped pruning are, however, poorly understood. Here, we provide evidence that semaphorins, activating the Plexin-A3 receptor, function as retraction inducers to trigger-stereotyped pruning of specific hippocampal mossy fiber and pyramidal axon branches. Both pruning events are defective in Plexin-A3 mutants, reflecting a cell-autonomous requirement for Plexin-A3. The distribution of mRNAs for Sema3F and Sema3A makes them candidates for triggering the pruning. In vitro, hippocampal neurons respond to semaphorins by retracting axon branches. These results implicate semaphorins as retraction inducers controlling stereotyped pruning in the mammalian brain. PMID- 12732139 TI - Links between tumor suppressors: p53 is required for TGF-beta gene responses by cooperating with Smads. AB - The p53 tumor suppressor belongs to a family of proteins that sense multiple cellular inputs to regulate cell proliferation, apoptosis, and differentiation. Whether and how these functions of p53 intersect with the activity of extracellular growth factors is not understood. Here, we report that key cellular responses to TGF-beta signals rely on p53 family members. During Xenopus embryonic development, p53 promotes the activation of multiple TGF-beta target genes. Moreover, mesoderm differentiation is inhibited in p53-depleted embryos. In mammalian cells, the full transcriptional activation of the CDK inhibitor p21(WAF1) by TGF-beta requires p53. p53-deficient cells display an impaired cytostatic response to TGF-beta signals. Smad and p53 protein complexes converge on separate cis binding elements on a target promoter and synergistically activate TGF-beta induced transcription. p53 can physically interact in vivo with Smad2 in a TGF-beta-dependent fashion. The results unveil a previously unrecognized link between two primary tumor suppressor pathways in vertebrates. PMID- 12732140 TI - Switch-of-function mutants based on morphology classification of Ras superfamily small GTPases. AB - Signaling proteins from the same family can have markedly different roles in a given cellular context. Here, we show that expression of one hundred constitutively active human small GTPases induced cell morphologies that fell into nine distinct classes. We developed an algorithm for pairs of classes that predicted amino acid positions that can be exchanged to create mutants with switched functionality. The algorithm was validated by creating switch-of function mutants for Rac1, CDC42, H-Ras, RalA, Rap2B, and R-Ras3. Contrary to expectations, the relevant residues were mostly outside known interaction surfaces and were structurally far apart from each other. Our study shows that specificity in protein families can be explored by combining genome-wide experimental functional classification with the creation of switch-of-function mutants. PMID- 12732141 TI - Actin dynamics control SRF activity by regulation of its coactivator MAL. AB - Rho GTPases regulate the transcription factor SRF via their ability to induce actin polymerization. SRF activity responds to G actin, but the mechanism of this has remained unclear. We show that Rho-actin signaling regulates the subcellular localization of the myocardin-related SRF coactivator MAL, rearranged in t(1;22)(p13;q13) AML. The MAL-SRF interaction displays the predicted properties of a Rho-regulated SRF cofactor. MAL is predominantly cytoplasmic in serum starved cells, but accumulates in the nucleus following serum stimulation. Activation of the Rho-actin signaling pathway is necessary and sufficient to promote MAL nuclear accumulation. MAL N-terminal sequences, including two RPEL motifs, are required for the response to signaling, while other regions mediate its nuclear export (or cytoplasmic retention) and nuclear import. MAL associates with unpolymerized actin through its RPEL motifs. Constitutively cytoplasmic MAL derivatives interfere with MAL redistribution and Rho-actin signaling to SRF. MAL associates with several SRF target promoters regulated via the Rho-actin pathway. PMID- 12732142 TI - Molecular basis of phosphorylation-induced activation of the NADPH oxidase. AB - The multi-subunit NADPH oxidase complex plays a crucial role in host defense against microbial infection through the production of reactive oxygen species. Activation of the NADPH oxidase requires the targeting of a cytoplasmic p40-p47 p67(phox) complex to the membrane bound heterodimeric p22-gp91(phox) flavocytochrome. This interaction is prevented in the resting state due to an auto-inhibited conformation of p47(phox). The X-ray structure of the auto inhibited form of p47(phox) reveals that tandem SH3 domains function together to maintain the cytoplasmic complex in an inactive form. Further structural and biochemical data show that phosphorylation of p47(phox) activates a molecular switch that relieves the inhibitory intramolecular interaction. This permits p47(phox) to interact with the cytoplasmic tail of p22(phox) and initiate formation of the active, membrane bound enzyme complex. PMID- 12732143 TI - The ubiquitin ligase activity in the DDB2 and CSA complexes is differentially regulated by the COP9 signalosome in response to DNA damage. AB - Nucleotide excision repair (NER) is a major cellular defense against the carcinogenic effects of ultraviolet light from the sun. Mutational inactivation of NER proteins, like DDB and CSA, leads to hereditary diseases such as xeroderma pigmentosum (XP) and Cockayne syndrome (CS). Here, we show that DDB2 and CSA are each integrated into nearly identical complexes via interaction with DDB1. Both complexes contain cullin 4A and Roc1 and display ubiquitin ligase activity. They also contain the COP9 signalosome (CSN), a known regulator of cullin-based ubiquitin ligases. Strikingly, CSN differentially regulates ubiquitin ligase activity of the DDB2 and CSA complexes in response to UV irradiation. Knockdown of CSN with RNA interference leads to defects in NER. These results suggest that the distinct UV response of the DDB2 and CSA complexes is involved in diverse mechanisms of NER. PMID- 12732144 TI - Closing the folding chamber of the eukaryotic chaperonin requires the transition state of ATP hydrolysis. AB - Chaperonins use ATPase cycling to promote conformational changes leading to protein folding. The prokaryotic chaperonin GroEL requires a cofactor, GroES, which serves as a "lid" enclosing substrates in the central cavity and confers an asymmetry on GroEL required for cooperative transitions driving the reaction. The eukaryotic chaperonin TRiC/CCT does not have such a cofactor but appears to have a "built-in" lid. Whether this seemingly symmetric chaperonin also operates through an asymmetric cycle is unclear. We show that unlike GroEL, TRiC does not close its lid upon nucleotide binding, but instead responds to the trigonal bipyramidal transition state of ATP hydrolysis. Further, nucleotide analogs inducing this transition state confer an asymmetric conformation on TRiC. Similar to GroEL, lid closure in TRiC confines the substrates in the cavity and is essential for folding. Understanding the distinct mechanisms governing eukaryotic and bacterial chaperonin function may reveal how TRiC has evolved to fold specific eukaryotic proteins. PMID- 12732145 TI - Alternative tertiary structure of tRNA for recognition by a posttranscriptional modification enzyme. AB - Transfer RNA (tRNA) canonically has the clover-leaf secondary structure with the acceptor, D, anticodon, and T arms, which are folded into the L-shaped tertiary structure. To strengthen the L form, posttranscriptional modifications occur on nucleotides buried within the core, but the modification enzymes are paradoxically inaccessible to them in the L form. In this study, we determined the crystal structure of tRNA bound with archaeosine tRNA-guanine transglycosylase, which modifies G15 of the D arm in the core. The bound tRNA assumes an alternative conformation ("lambda form") drastically different from the L form. All of the D-arm secondary base pairs and the canonical tertiary interactions are disrupted. Furthermore, a helical structure is reorganized, while the rest of the D arm is single stranded and protruded. Consequently, the enzyme precisely locates the exposed G15 in the active site, by counting the nucleotide number from G1 to G15 in the lambda form. PMID- 12732146 TI - Program-specific distribution of a transcription factor dependent on partner transcription factor and MAPK signaling. AB - Specialized gene expression programs are induced by signaling pathways that act on transcription factors. Whether these transcription factors can function in multiple developmental programs through a global switch in promoter selection is not known. We have used genome-wide location analysis to show that the yeast Ste12 transcription factor, which regulates mating and filamentous growth, is bound to distinct program-specific target genes dependent on the developmental condition. This condition-dependent distribution of Ste12 requires concurrent binding of the transcription factor Tec1 during filamentation and is differentially regulated by the MAP kinases Fus3 and Kss1. Program-specific distribution across the genome may be a general mechanism by which transcription factors regulate distinct gene expression programs in response to signaling. PMID- 12732149 TI - Acceleration of cure or optimisation of care? PMID- 12732147 TI - A global control region defines a chromosomal regulatory landscape containing the HoxD cluster. AB - During limb development, coordinated expression of several Hoxd genes is required in presumptive digits. We searched for the underlying control sequences upstream from the cluster and found Lunapark (Lnp), a gene which shares limb and CNS expression specificities with both Hoxd genes and Evx2, another gene located nearby. We used a targeted enhancer-trap approach to identify a DNA segment capable of directing reporter gene expression in both digits and CNS, following Lnp, Evx2, and Hoxd-specific patterns. This DNA region showed an unusual interspecies conservation, including with its pufferfish counterpart. It contains a cluster of global enhancers capable of controlling transcription of several genes unrelated in structure or function, thus defining large regulatory domains. These domains were interrupted in the Ulnaless mutation, a balanced inversion that modified the topography of the locus. We discuss the heuristic value of these results in term of locus specific versus gene-specific regulation. PMID- 12732150 TI - Will IARC election help to change "name security" policy? PMID- 12732153 TI - UK launch "virtual" tumour bank to improve treatment research. PMID- 12732157 TI - AACR cancels annual meeting because of SARS. PMID- 12732158 TI - India cuts budget for cancer treatment. PMID- 12732160 TI - Family history reveals lung-cancer risk. PMID- 12732162 TI - Predicting outcome and tailoring treatment in high-grade gliomas. PMID- 12732163 TI - Total gastrectomy: options for the restoration of gastrointestinal continuity. AB - Total gastric resection and the subsequent selection of the optimum procedure for reconstruction of the oesophageal-intestinal tract is a highly debated topic. Many different types of reconstruction have been proposed, but attempts to objectively validate the potential advantages of these techniques of have not produced any definitive conclusions. The main aim of reconstruction should be to maintain the nutritional status and quality of life of the patient, with the additional goal of achieving similar function to normal gut. In this review, we discuss the prospectively randomised studies of gastrointestinal reconstruction that were done between 1987 and 2002. PMID- 12732164 TI - Intraperitoneal antineoplastic drug delivery: rationale and results. AB - The use of intraperitoneal drug delivery in the treatment of malignant disease confined to the peritoneal cavity is based on the theoretical potential for increased exposure of the tumour to antineoplastic agents leading to improved cytotoxicity. Phase I studies have explored the safety and pharmacokinetic advantage of the regional administration of several drugs, including cisplatin (10 times higher than systemic delivery) and paclitaxel (1000 times higher). Phase II trials of second-line intraperitoneal chemotherapy of ovarian cancer, generally with cisplatin, have shown the potential for patients to achieve surgically-documented complete responses. Randomised trials of second-line regional therapy in patients with ovarian cancer have yet to be conducted, although non-randomised single institution experience has suggested the potential for long-term disease-free survival with this strategy. By contrast, several well planned randomised trials of first-line intraperitoneal chemotherapy of small volume residual advanced ovarian cancer after primary surgical cytoreduction have reported a survival advantage with regional drug delivery. Although a rationale can be proposed for intraperitoneal antineoplastic drug delivery in non-ovarian malignant disease involving the peritoneal cavity, current data do not support the use of this strategy outside the confines of well-designed clinical trials. PMID- 12732165 TI - Clinical picture: Primary cardiac synovial sarcoma. PMID- 12732166 TI - Aetiology of bone disease and the role of bisphosphonates in multiple myeloma. AB - Osteolytic bone disease is a major cause of morbidity in patients with multiple myeloma. Our understanding of the pathophysiology of multiple myeloma has increased substantially during the past decade. However the underlying mechanisms of bone destruction and the treatments available have, until recently, received relatively little specific attention. In this review, we provide an overview of the RANK/RANKL/osteoprotegerin system; we describe its interaction with other cellular mechanisms, through which malignant plasma cells drive osteolysis, and explain how bisphosphonates can be used to block this action. We also review the supporting evidence for bisphosphonates as the treatment of choice for patients with bone complications related to multiple myeloma, and discuss possible developments for targeted therapy in the future. PMID- 12732167 TI - Management of multiple myeloma: a systematic review and critical appraisal of published studies. AB - We have done a systematic review of all randomised studies in myeloma, identified through a comprehensive search. Our aim was to investigate and critically examine the effects of various treatment modalities on outcome in patients with multiple myeloma and address 22 specific clinical questions in the management of this disease. As a result of our analysis we identified two therapeutic advances in the management of myeloma that, according to the evidence, are most important for improving outcome. These advances were: introduction of high dose chemotherapy, which appears to be superior to conventional chemotherapy, and the use of bisphosphonates, which decrease the probability of pathological vertebral fractures. However, the overall quality of the body of evidence for myeloma management was poor. Many trials were done with small sample sizes, and did not include reporting power analysis. The majority of studies had inadequate allocation concealment, and few were analysed according to intention to treat principle. We conclude that the quality of total evidence supporting treatment recommendations in myeloma is modest at best and has an ample scope for improvement. PMID- 12732168 TI - Statistical design in phase II clinical trials and its application in breast cancer. AB - Several statistical designs for phase II studies have been proposed, but they are frequently misunderstood or not applied at all. In this review we describe the major characteristics of the available designs. To investigate the extent to which statistical designs were used in some recent phase II studies, and which designs were the most common, we did a survey of 145 trials involving treatment of breast cancer. Studies selected for the survey were published between 1995 and 1999 in one of seven specific oncology journals (all with impact factor consistently higher than 2). 94 of the studies (64.8%) did not have an identifiable statistical design. However, among the 51 studies with statistical design there was a notable heterogeneity in the type of design applied. We put together a list of factors associated with use of statistical design at univariate analysis. These factors included: referral to a previous phase I study, recent trial start date, private sponsorship, single-agent treatment, and multicentre organisation. Single-agent treatment (OR 2.35; 95% CI 1.01-5.51) and multicentre organisation (OR 3.24; 95% CI 1.47-7.15) were independently predictive of the presence of statistical design. Publication in journals with high impact factors and short intervals between the start of the study and publication were also correlated with statistical design. PMID- 12732169 TI - The use of opioids and sedatives at the end of life. AB - Opioids and sedative drugs are commonly used to control symptoms in patients with advanced cancer. However, it is often assumed that the use of these drugs inevitably results in shortening of life. Ethically, this outcome is excused by reference to the doctrine of double effect. In this review, we assess the evidence for patterns of use of opioids and sedatives in palliative care and examine whether the doctrine of double effect is needed to justify their use. We conclude that patients are more likely to receive higher doses of both opioids and sedatives as they get closer to death. However, there is no evidence that initiation of treatment, or increases in dose of opioids or sedatives, is associated with precipitation of death. Thus, we conclude that the doctrine of double effect is not essential for justification of the use of these drugs, and may act as a deterrent to the provision of good symptom control. PMID- 12732170 TI - Carcinoma of the gallbladder with unusual umbilical metastasis. PMID- 12732176 TI - Structural and functional characterization of Runx1, CBF beta, and CBF beta SMMHC. AB - Core binding factors (CBFs) are heterodimeric transcription factors consisting of a DNA-binding CBF alpha subunit and non-DNA-binding CBF beta subunit. DNA binding and heterodimerization is mediated by a single domain in the CBF alpha subunit called the Runt domain, while sequences flanking the Runt domain confer other biochemical activities such as transactivation. On the other hand, the heterodimerization domain in CBF beta is the only functional domain that has been identified in this subunit. The biophysical properties of the Runt domain and the CBF beta heterodimerization domain, and the structures of the isolated domains as well as of the Runt domain-DNA, Runt domain-CBF beta, and ternary Runt domain-CBF beta-DNA complexes, have been characterized over the past several years, and are summarized in this review. PMID- 12732177 TI - Runx3 is essential for the target-specific axon pathfinding of trkc-expressing dorsal root ganglion neurons. AB - Dorsal root ganglion (DRG) neurons project their axons to specific target layers in the gray matter of the spinal cord, according to their sensory modality (Neuron 30 (2001), 707; Cell 101 (2000), 485; Neuron 31 (2001), 59; J. Comp. Neurol. 380 (1997), 215; Sensory Neurons, Oxford Univ. Press, New York, 1992, p. 131). Expression of runt-related Runx/AML genes (Mech. Dev. 109 (2001), 413) on subtypes of DRG neurons suggests their involvement in lamina-specific afferent differentiation and maturation. Here we show that Runx3-/- mice display severe limb ataxia and abnormal posture and that most of them die shortly after birth. They show that proprioceptive afferent axons fail to reach the ventral horn and have a smaller dorsal funiculus in their spinal cords. Despite the strong resemblance of this phenotype to that of knockout mice deficient in neurotrophin 3 (NT-3) (Cell 77 (1994), 503; Nature 369 (1994), 658) and its receptor, trkC, (Nature 368 (1994), 249), which show proprioceptive afferent loss through selective neuronal cell death, Runx3-/- mice maintain normal number of TrkC/trkC positive DRG neurons throughout development. Our results suggest that Runx3 controls the target-specific axon pathfinding of trkC-expressing DRG neurons in the spinal cord. PMID- 12732178 TI - Phylogenesis and regulated expression of the RUNT domain transcription factors RUNX1 and RUNX3. AB - The RUNX transcription factors are key regulators of lineage specific gene expression in developmental pathways. The mammalian RUNX genes arose early in evolution and maintained extensive structural similarities. Sequence analysis suggested that RUNX3 is the most ancient of the three mammalian genes, consistent with its role in neurogenesis of the monosynaptic reflex arc, the simplest neuronal response circuit, found in Cnidarians, the most primitive animals. All RUNX proteins bind to the same DNA motif and act as activators or repressors of transcription through recruitment of common transcriptional modulators. Nevertheless, analysis of Runx1 and Runx3 expression during embryogenesis revealed that their function is not redundant. In adults both Runx1 and Runx3 are highly expressed in the hematopoietic system. At early embryonic stages we found strong Runx3 expression in dorsal root ganglia neurons, confined to TrkC sensory neurons. In the absence of Runx3, knockout mice develop severe ataxia due to the early death of the TrkC neurons. Other phenotypic defects of Runx3 KO mice including abnormalities in thymopoiesis are also being investigated. PMID- 12732179 TI - Cbf beta is involved in maturation of all lineages of hematopoietic cells during embryogenesis except erythroid. AB - The transcription factor Cbf beta forms a heterodimeric complex with members of the Runx family of proteins. Together, Cbf beta and Runx1 play a critical role in the establishment of definitive hematopoiesis in mouse embryos. Previously, we used a Cbfb-GFP "knock-in" mouse model to demonstrate that Cbf beta is expressed in hematopoietic stem cells of the mouse fetal liver and aorta-gonad-mesonephros (Blood 100 (2002), 2449). We also examined the expression pattern of Cbf beta in different lineages of adult hematopoietic cells and found that it is expressed uniformly in all lineages except B lymphocytes and erythroid cells. Cbf beta expression decreases during maturation of B cells in the adult bone marrow, and is not expressed in nucleated erythroid precursors. Here, we examine the expression of Cbf beta in various hematopoietic lineages, including myeloid, lymphoid, and erythroid during late stages of embryonic development, and compare it to the pattern observed in adults. We find that there are subtle differences in expression of Cbf beta-GFP in embryonic hematopoietic cells compared to their adult counterparts, but that the overall pattern is the same. Our data complement recently published data on hematopoetic defects observed in transgenic Cbfb-null mouse embryos partially rescued by ectopic expression of Cbfb (Nature Genet. 32 (2002), 633; Nature Genet. 32 (2002), 645). and supports the emerging view that Cbf beta and Runx proteins are required for normal maturation of hematopoietic cells as well as establishment of definitive hematopoiesis. PMID- 12732180 TI - Intranuclear organization of RUNX transcriptional regulatory machinery in biological control of skeletogenesis and cancer. AB - RUNX (AML/CBFA/PEBP2) transcription factors serve as paradigms for obligatory relationships between nuclear structure and physiological control of phenotypic gene expression. The RUNX proteins contribute to tissue restricted transcription by sequence-specific binding to promoter elements of target genes and serving as scaffolds for the assembly of coregulatory complexes that mediate biochemical and architectural control of activity. We will present an overview of approaches we are pursuing to address: (1) the involvement of RUNX proteins in governing competency for protein/DNA and protein/protein interactions at promoter regulatory sequences; (2) the recruitment of RUNX factors to subnuclear sites where the machinery for expression or repression of target genes is organized; and (3) the trafficking and integration of regulatory signals that control RUNX mediated transcription. PMID- 12732181 TI - The t(8;21) fusion protein contacts co-repressors and histone deacetylases to repress the transcription of the p14ARF tumor suppressor. AB - The t(8;21) is one of the most frequent chromosomal translocations associated with acute leukemia. The translocation fuses the DNA binding domain of AML1 to nearly all of the ETO co-repressor. ETO associates with the mSin3 and N-CoR co repressors as well as histone deacetylases 1, 2, and 3. Although this is one of the most frequent chromosomal translocations in acute leukemia, accounting for 10 15% of the cases of acute myeloid leukemia (AML), the direct targets for transcriptional regulation that stimulate leukemogenesis are unknown. We found that AML1-ETO repressed the promoter of p14(ARF) tumor suppressor in transient transfection assays and reduced endogenous levels of p14(ARF) expression in multiple cell types. Chromatin immunoprecipitation assays demonstrated that AML1 ETO bound to the p14(ARF) promoter. In acute myeloid leukemia samples containing the t(8;21), levels of p14(ARF) mRNA were markedly lower when compared to other acute myeloid leukemias. Therefore, p14(ARF) is a direct transcriptional target of AML1-ETO. PMID- 12732182 TI - Functional analysis of RUNX2 mutations in cleidocranial dysplasia: novel insights into genotype-phenotype correlations. AB - Cleidocranial dysplasia (CCD) is an inherited autosomal-dominant skeletal disease caused by heterozygous mutations in the osteoblast-specific transcription factor, RUNX2. We have performed mutational analysis of RUNX2 on 24 unrelated patients with CCD. In 17 patients, 16 distinct mutations were detected in the coding region of RUNX2: 4 frameshift, 3 nonsense, 6 missense, and 2 splicing mutations alongside one polymorphism. The missense mutations were all clustered within the Runt domain and their protein products showed neither DNA binding nor transactivation. On the other hand, some mutant RUNX2 had the Runt domain intact and remained partially competent for transactivation. Coincidentally, one important phenotype of CCD, the short stature, was significantly milder in the patients with the intact Runt domain than those without. Furthermore, a remarkable correlation was found between the short stature and the number of supernumerary teeth. On the other hand, the classic CCD phenotype, hypoplastic clavicles or open fontanelles, was invariably observed regardless of the degree of short stature or supernumerary teeth. Overall, these results suggest that CCD could result from a much smaller loss in the RUNX2 function than envisioned on the basis of the conventional haploinsufficiency model. This makes an interesting contrast to the case of familial and sporadic leukemias mediated by RUNX1 mutations, in which mutants acting in a dominant negative manner have been suggested to confer a higher propensity to develop leukemia. PMID- 12732184 TI - The ETO domain is necessary for the developmental abnormalities associated with AML1-ETO expression in the hematopoietic stem cell compartment in vivo. AB - Translocation of the ETO gene on human chromosome 8 with the AML1 gene on chromosome 21 (AML1-ETO) is a recurrent cytogenetic abnormality associated with approximately 12% of acute myelogenous leukemia (AML) cases. To understand the contribution of the t(8;21) to AML, we transduced purified hematopoietic stem cells (HSC) with a retroviral vector that coexpressed AML1-ETO or just the AML1 portion (AML1d) of the translocation along with a green fluorescent protein reporter gene. Animals reconstituted with AML1-ETO-expressing cells exhibited many of the hematopoietic developmental abnormalities seen in the bone marrow of human patients with the t(8;21), although the animals did not develop acute leukemia. We noted a gradual increase in primitive myeloblasts that accounted for approximately 10% of bone marrow by 10 months posttransplant. Consistent with this observation was a 50-fold increase in myeloid colony-forming cells in vitro. In addition, accumulation of late stage metamyelocytes was observed in bone marrow along with an increase in immature eosinophil myelocytes that showed abnormal basophilic granulation. There was also a gradual increase in both the frequency and absolute number of AML1-ETO-expressing HSC so that by 10 months posttransplant, there were 29-fold greater HSC numbers than in transplant-matched control mice. These phenotypes were not observed in animals reconstituted with cells expressing only the DNA-binding domain of AML1, suggesting that the ETO domain is necessary to establish the developmental abnormalities associated with AML1-ETO expression in HSC. PMID- 12732183 TI - The Runx genes as dominant oncogenes. AB - We have shown previously that Runx2 is a frequent target (approximately equal to 30%) for proviral insertion in murine leukemia virus (MLV) induced T cell tumors in CD2-MYC transgenic mice. Further investigation of a large panel of these tumors revealed that a small number also contain insertions at either Runx3 or Runx1. None of the tumors contained insertions at more than one family member, but in each case proviral insertion was associated with a high level of expression from the upstream (P1) promoter of the respective target gene. Moreover, we confirmed that transcriptional activation of Runx1 does not affect the integrity of the coding sequence, as previously observed for Runx2. These observations suggest that the three Runx genes act as functionally redundant oncogenes in T-cell lymphoma development. To explore the oncogenic potential of Runx2 further we created transgenic mice that over-express this gene in the T cell compartment. These CD2-Runx2 animals show a preneoplastic enlargement of the CD8 immature single positive (ISP) thymocyte pool and develop lymphomas at a low incidence. Although the CD8 ISP population is greatly increased, unlike their wild type counterparts these cells are largely non-cycling. Co-expression of c MYC in this lineage accentuates the CD8 ISP skew and induces rapid tumor development, confirming the potent synergy that exists between these two oncogenes. Experiments designed to understand the nature of the observed synergy are ongoing and are based on the hypothesis that Runx2 may exert a survival effect in c-MYC expressing tumors in vivo while c-MYC may rescue cells from the antiproliferative effects of Runx2. The oncogenic potential of Runx1 is also being assessed using primary murine embryonic fibroblasts (MEFs). These studies have revealed that while Runx1 exerts a growth suppressive effect in wild type cells a growth promoting effect is seen in the absence of p53, suggesting that the Runx genes may harbor latent oncogene-like properties. PMID- 12732185 TI - A DNA-binding-independent pathway of repression by the Drosophila Runt protein. AB - DNA-binding proteins are important for regulating gene expression during development. It is widely assumed that this regulation involves sequence-specific DNA binding by these transcription factors to cognate cis-regulatory sequences of their downstream target genes. However, studies in both the Drosophila and the mouse model systems have provided examples in which the DNA-binding activity of a transcription factor is not essential for in vivo function. Using a system that allows for quantitative analysis of gene function in the Drosophila embryo, we have discovered a DNA-binding-independent activity of Runt, the founding member of the RUNX family of transcriptional regulators. Examination of the in vivo potency of a DNA-binding-defective form of Runt reveals differential requirements for DNA binding in the regulation of different downstream target genes. DNA binding is not required for establishing repression of the odd-numbered stripes of the segment polarity gene engrailed, but does contribute to Runt's role as a regulator of sloppy-paired, another downstream target gene in the pathway of segmentation. We investigate this DNA-binding-independent pathway using a genetic screen for dose-dependent modifiers of runt activity. These studies reveal that DNA-binding proteins encoded by the tramtrack locus cooperate with Runt to repress engrailed. These results provide new insights into the context-dependent regulatory functions of Runt domain proteins and provide a paradigm for understanding DNA-binding-independent regulation by developmentally important transcription factors. PMID- 12732186 TI - Transcriptional regulation of hematopoiesis in Drosophila. AB - As in mammals, blood cells in Drosophila are derived from a common multipotent hematopoietic precursor population. In the embryo, these precursors are derived from the head mesoderm, whereas larval hematopoietic precursors are found in a specialized organ called the lymph gland. This shift in location of hematopoietic differentiation is reminiscent of similar events that occur during mammalian development. Recent analysis has identified several transcriptional regulators in Drosophila that influence hematopoietic lineage commitment. Interestingly, many of these factors are similar to factors directing mammalian hematopoietic differentiation. Although Drosophila blood cells are much less varied in terms of specific lineages, it would appear that many mechanistic aspects by which hematopoietic cell fate is determined have been conserved between Drosophila and mammals. Herein, we describe the Drosophila blood cell types, their physical origin, and the transcriptional regulators that govern this process. PMID- 12732187 TI - Next-generation protein-handling method: puromycin analogue technology. AB - Puromycin is a well-known antibiotic that inhibits protein synthesis by competitive incorporation against an aminoacyl tRNA on the ribosome A site. Novel technology using this property of puromycin has been developed for convenient handling methods in protein research. Puromycin modified with another molecule is incorporated into a protein at the C-terminus, thus linking the desired molecule to the protein. Combination of in vitro translation with puromycin analogues has resulted in novel technologies such as display technology for screening, fluorescence labeling, affinity purification, and protein chip for proteomics. PMID- 12732188 TI - Decorin modulates matrix mineralization in vitro. AB - Decorin (DCN), a member of small leucine-rich proteoglycans, is known to modulate collagen fibrillogenesis. In order to investigate the potential roles of DCN in collagen matrix mineralization, several stable osteoblastic cell clones expressing higher (sense-DCN, S-DCN) and lower (antisense-DCN, As-DCN) levels of DCN were generated and the mineralized nodules formed by these clones were characterized. In comparison with control cells, the onset of mineralization by S DCN clones was significantly delayed; whereas it was markedly accelerated and the number of mineralized nodules was significantly increased in As-DCN clones. The timing of mineralization was inversely correlated with the level of DCN synthesis. In these clones, the patterns of cell proliferation and differentiation appeared unaffected. These results suggest that DCN may act as an inhibitor of collagen matrix mineralization, thus modulating the timing of matrix mineralization. PMID- 12732189 TI - Efficient removal of LoxP-flanked genes by electroporation of Cre-recombinase mRNA. AB - Introduction of Cre-recombinase in target cells is currently achieved by transfection of plasmid DNA or by viral-mediated transduction. However, efficiency of non-viral DNA transfection is often low in many cell types, and the use of viral vectors for transduction implies a more complex and laborious manipulation associated with safety issues. We have developed a non-viral non-DNA technique for rapid and highly efficient excision of LoxP-flanked DNA sequences based on electroporation of in vitro transcribed mRNA encoding Cre-recombinase. A K562-DSRed[EGFP] cell line was developed in order to measure Cre-mediated recombination by flow cytometric analysis. These cells have a stable integrated DSRed reporter gene flanked by two LoxP sites, and an EGFP reporter gene, which could only be transcribed when the coding sequence for DSRed was removed. The presented data show recombination efficiencies, as measured by appearance of EGFP fluorescence, of up to 85% in Cre-recombinase mRNA-electroporated K562 DSRed[EGFP] cells. In conclusion, mRNA electroporation of Cre-recombinase is a powerful, safe, and clinically applicable alternative to current technologies used for excision of stably integrated LoxP-flanked DNA sequences. PMID- 12732190 TI - Myosin light chain kinase stimulates smooth muscle myosin ATPase activity by binding to the myosin heads without phosphorylating the myosin light chain. AB - Myosin light chain kinase (MLCK) is a multifunctional regulatory protein of smooth muscle contraction [IUBMB Life 51 (2001) 337, for review]. The well established mode for its regulation is to phosphorylate the 20 kDa myosin light chain (MLC 20) to activate myosin ATPase activity. MLCK exhibits myosin-binding activity in addition to this kinase activity. The myosin-binding activity also stimulates myosin ATPase activity without phosphorylating MLC 20 [Proc. Natl. Acad. Sci. USA 96 (1999) 6666]. We engineered an MLCK fragment containing the myosin-binding domain but devoid of a catalytic domain to explore how myosin is stimulated by this non-kinase pathway. The recombinant fragment thus obtained stimulated myosin ATPase activity by V(max)=5.53+/-0.63-fold with K(m)=4.22+/ 0.58 microM (n=4). Similar stimulation figures were obtained by measuring the ATPase activity of HMM and S1. Binding of the fragment to both HMM and S1 was also verified, indicating that the fragment exerts stimulation through the myosin heads. Since S1 is in an active form regardless of the phosphorylated state of MLC 20, we conclude that the non-kinase stimulation is independent of the phosphorylating mode for activation of myosin. PMID- 12732192 TI - Genomic overview of serine proteases. AB - Serine proteases (SP) are peptidases with a uniquely activated serine residue in the substrate-binding pocket. They represent about 0.6% of all proteins in the human genome. SP are involved in many vital functions such as digestion, blood clotting, fibrinolysis, fertilization, and complement activation and are related to many diseases including cancer, arthritis, and emphysema. In this study, we performed a genomic analysis of human serine proteases utilizing different databases, primarily that of MEROPS. SP are distributed along all human chromosomes except 18 and Y with the highest density (23 genes) on chromosome 19. They are either randomly located within the genome or occur in clusters. We identified a number of SP clusters, the largest being the kallikrein cluster on chromosome 19q13.4 which is formed of 15 adjacent genes. Other clusters are located on chromosomes 19p13, 16p13, 14q11, 13q35, 11q22, and 7q35. Genes of each cluster tend to be of comparable sizes and to be transcribed in the same direction. The members of some clusters are sometimes functionally related, e.g., the involvement of many kallikreins in endocrine-related malignancies and the hematopoietic cluster on chromosome 14. It is hypothesized that members of some clusters are under common regulatory mechanisms and might be involved in cascade enzymatic pathways. Several functional domains are found in SP, which reflect their functional diversity. Membrane-type SP tend to cluster in 3 chromosomes and have some common structural domains. Several databases are available for screening, structural and functional analysis of serine proteases. With the near completion of the Human Genome Project, research will be more focused on the interactions between SP and their involvement in pathophysiological processes. PMID- 12732191 TI - Lipoprotein lipase mRNA in white adipose tissue but not in skeletal muscle is increased by pioglitazone through PPAR-gamma. AB - Lipoprotein lipase (LPL), a key enzyme for triglyceride hydrolysis, is an insulin dependent enzyme and mainly synthesized in white adipose tissue (WAT) and skeletal muscles (SM). To explore how pioglitazone, an enhancer of insulin action, affects LPL synthesis, we examined the effect of pioglitazone on LPL mRNA levels in WAT or SM of brown adipose tissue (BAT)-deficient mice, which develop insulin resistance and hypertriglyceridemia. Both LPL mRNA of WAT and SM were halved in BAT-deficient mice. Pioglitazone increased LPL mRNA in WAT by 8-fold, which was substantially associated with a 4-fold increase of peroxisome proliferator activated receptor (PPAR)-gamma mRNA (r=0.97, p<0.0001), whereas pioglitazone did not affect LPL mRNA in SM. These results suggest that pioglitazone exclusively increases LPL production in WAT via stimulation of PPAR gamma synthesis. Since pioglitazone does not affect LPL production in SM, this would contribute to prevent the development of insulin resistance due to lipotoxicity. PMID- 12732193 TI - Production, purification, and functional analysis of recombinant human and mouse 17beta-hydroxysteroid dehydrogenase type 7. AB - 17beta-Hydroxysteroid dehydrogenases (17HSDs) have a central role in the regulation of the biological activity of sex steroid hormones. There is increasing evidence that in addition to their importance in gonads, these hormones also have substantial metabolic roles in a variety of peripheral tissues. In the present study, the cDNA of human 17HSD type 7 was cloned. In silico, the gene corresponding to the cDNA was localized on chromosome 1q23, close to the locus of hereditary prostate cancer 1 (HPC1) (1q24-25) and primary open-angle glaucoma (GLC1A) (1q23-25). Further, a pseudogene was found on chromosome 1q44, close to the locus of predisposing for early-onset prostate cancer (PCaP) (1q42.2-43). Both human (h17HSD7) and mouse 17HSD type 7 (m17HSD7) were for the first time produced as recombinant proteins and purified for functional analyses. Further, kinetic parameters and specific activities were described. h17HSD7 converted estrone (E1) to a more potent estrogen, estradiol (E2), and dihydrotestosterone (DHT), a potent androgen, to an estrogenic metabolite 5alpha-androstane-3beta, 17beta-diol (3betaA-diol) equally, thereby catalyzing the reduction of the keto group in either 17- or 3-position of the substrate. Minor 3betaHSD-like activity towards progesterone (P) and 20 hydroxyprogesterone (20-OH-P), leading to the inactivation of P by h17HSD7, was also detected. m17HSD7 efficiently catalyzed the reaction from E1 to E2 and moderately converted DHT to an inactive metabolite 5alpha-androstane 3alpha,17beta-diol (3alphaA-diol) and to an even lesser degree 3betaA-diol. The mouse enzyme did not metabolize P or 20-OH-P. The expression of 17HSD type 7 was observed widely in human tissues, most distinctly in adrenal gland, liver, lung, and thymus. Based on the enzymatic characteristics and tissue distribution, we conclude that h17HSD7 might be an intracrine regulator of steroid metabolism, fortifying the estrogenic milieu in peripheral tissues. PMID- 12732194 TI - Hepatic lineages isolated from developing rat liver show different ways of maturation. AB - Immunocytochemical analysis revealed that different hepatic cell types exist during liver development: (i). cells co-expressing the stem-cell marker Thy1 and the hepatic lineage marker CK-18 and (ii). cells only expressing CK-18 (hepatoblasts). In this study we separated the different hepatic cells and analyzed gene-expression and phenotype. Fetal rat livers were digested by collagenase solution. OX43- and OX44-positive hematopoietic cells were depleted and Thy1-positive cells were enriched using Magnetic cell sorting. The different cell compartments were analyzed by RT-PCR and immunocytochemistry for Thy1, CK 18, AFP, and albumin. Hepatoblasts expressed albumin at all times and AFP in the early stages. Thy1-enriched cells expressed CK-18 at all times, albumin in the early, and AFP in the late stages. Thy1-positive cells from fetal livers express liver specific genes. The data suggest that Thy1-positive hepatic cells develop towards hepatic stem cells, and hepatoblasts develop towards mature hepatocytes of the adult liver. PMID- 12732195 TI - cDNA microarray analysis reveals chop-10 plays a key role in Sertoli cell injury induced by bisphenol A. AB - We examined the time course of changes in gene expression in detail using cDNA microarray analysis of mouse Sertoli TTE3 cells treated with bisphenol A (BPA). A subtoxic dose of BPA (200 microM) transiently increased intracellular Ca(2+) concentration and time-dependently induced an increase in mRNA level of 78-kDa glucose-regulated protein, indicating that BPA induces endoplasmic reticulum stress. Of the 865 genes analyzed, 31 genes showed increased levels of expression. TaqMan analysis confirmed that the mRNA levels of chop-10, fra-2, c myc, and ornithine decarboxylase were increased, and showed that chop-10 is the most sensitive gene. The expression level of chop-10 protein and cell injury induced by BPA were significantly reduced in stable TTE3 cells overexpressing full-length chop-10 antisense RNA. We conclude that chop-10 plays a key role in Sertoli cell injury induced by BPA. PMID- 12732196 TI - Contribution of presenilin/gamma-secretase to calsenilin-mediated apoptosis. AB - Mutant presenilins cause early-onset of familial Alzheimer's disease and render cells vulnerable to apoptosis. Calsenilin/DREAM/KChIP3 is a multifunctional calcium-binding protein that interacts with presenilin and mediates calcium mediated apoptosis. In the present study, we report that the calsenilin-mediated apoptosis is regulated by presenilin. The expression of calsenilin was highly up regulated in neuronal cells undergoing Abeta42-triggered cell death. The incidence of calsenilin-mediated apoptosis was diminished in presenilin-1(-/-) mouse embryonic fibroblast cells or neuronal cells stably expressing a loss-of function presenilin-1 mutant. On the contrary, an array of familial Alzheimer's disease-associated presenilin mutants (gain-of-function) increased calsenilin induced cell death. Moreover, gamma-secretase inhibitors, including compound E and DAPT, decreased the calsenilin-induced cell death. These results suggest that the pro-apoptotic activity of calsenilin coordinates with presenilin/gamma secretase activity to play a crucial role in the neuronal death of Alzheimer's disease. PMID- 12732197 TI - Novel human G-protein-coupled receptors. AB - G-protein-coupled receptors (GPCRs) are important mediators of signal transduction and targets for pharmacological therapeutics. Novel receptor-ligand systems have been discovered through the identification and analysis of orphan GPCRs (oGPCRs). Here we describe the discovery of seven novel human genes encoding oGPCRs. Each novel oGPCR gene was discovered using customized searches of the GenBank genomic databases with previously known GPCR-encoding sequences. The expressed genes can now be used in assays to determine endogenous and pharmacological ligands. GPR133, GPR134, GPR135, GPR136, and GPR137 share identities with a prostate-specific odorant-like GPCR-encoding gene (PSGR). GPR138 and GPR139 share identities with an odorant-like gene derived from human erythroid cells. Transcripts encoding GPR133, GPR134, GPR135, GPR136, and GPR137 were detected in various CNS tissues. The expression of odorant-like genes in non olfactory tissues requires further clarification, which may be achieved through the search for endogenous cognate ligands for these and other oGPCRs. PMID- 12732198 TI - Different protofilament-dependence of the microtubule binding between MAP2 and MAP4. AB - To see a molecular basis of the difference in the microtubule binding between MAP2 and MAP4, we compared the binding of them onto microtubule and Zinc-sheet in the presence of various concentrations of NaCl. The Zinc-sheet is the lateral association of protofilaments arranged in an antiparallel fashion with alternatively exposed opposite surfaces, so that binding requiring adjacent protofilaments is restricted. While the salt-dependence of the MAP2 desorption was not altered between these tubulin polymers, MAP4 dissociated from Zinc-sheet at lower concentrations of NaCl than from microtubule. These results suggest that single protofilament is sufficient for microtubule binding of MAP2 as observed by Al-Bassam et al. [J. Cell Biol. 157 (2002) 1187], but MAP4 appeared to interact with adjacent protofilaments during microtubule-binding. Weakened binding on Zinc sheets was also observed in the projection domain-deletion mutants of MAP4, so that the difference in the protofilament-dependence would lie in the relatively conserved microtubule-binding domain. PMID- 12732199 TI - Single-molecule detection of efflux pump machinery in Pseudomonas aeruginosa. AB - Real-time single-molecule microscopy and spectroscopy were used to monitor single molecules moving in and out of live bacterial cells, Pseudomonas aeruginosa. Ethidium bromide (EtBr) was chosen as the fluorescence probe because it emitted a weak fluorescence in aqueous solution (outside of the cells) and became strongly fluorescent as it entered the cells and intercalated with DNA. Such changes in fluorescence intensity by individual EtBr molecules were measured to determine the influx and efflux rates of EtBr by the cells. The transport rates for EtBr through the energized extrusion pumps of these strains (WT, nalB-1, and DeltaABM) of P. aeruginosa were measured and showed stochastic behavior with the average being (2.86+/-0.12), (2.80+/-0.13), and (2.74+/-0.39) x s(-1), respectively. The transport rates of the three strains were independent of substrate concentration at the single-molecule level. In contrast to bulk (many molecules) measurements, single-molecule detection allowed the influx and efflux kinetics to be observed in low substrate concentrations at the molecular level. PMID- 12732200 TI - Reversible methionine sulfoxidation of Mycobacterium tuberculosis small heat shock protein Hsp16.3 and its possible role in scavenging oxidants. AB - Mycobacterium tuberculosis (TB) small heat shock protein Hsp16.3 was found to be a major membrane protein that is most predominantly expressed under oxidative stress and is localized to the thickened cell envelope. Gene knock-out studies indicate that the Hsp16.3 protein is required for TB to grow in its host macrophage cells. The physiological function of Hsp16.3 has not yet revealed. Our analyses via mass spectrometry, conformation-dependent trypsin digestion, nondenaturing pore gradient electrophoresis, ANS-binding fluorescence measurements, and circular dichroism demonstrate that the three and only the three methionine residues (cysteine and tryptophan residues, which can also be readily oxidized by such oxidant as H(2)O(2), are absent in Hsp16.3) can be readily sulfoxidized with H(2)O(2) treatment in vitro, and the methionine sulfoxide can be effectively reduced back to the methionine form. Interconversion between the methionine and methioninesulfoxide has been confirmed by selective oxidation and reduction. The sulfoxidation leads to a small degree of conformational change, which in turn results in a significant decrease of the chaperone-like activity. Data presented in this report strongly implicate that reversible sulfoxidation/desulfoxidation of methionine residues may occur in Hsp16.3, which serves as a way to scavenger reactive oxygen or nitrogen species abundantly present in macrophage cells, thus protecting the plasma membrane and other components of M. tuberculosis allowing their survival in such bacteriocidal hosts. PMID- 12732201 TI - L-cysteine administration prevents liver fibrosis by suppressing hepatic stellate cell proliferation and activation. AB - Recent studies showed that the function of some amino acids is not only nutritional but also pharmacological. However, the effects of amino acids on liver fibrosis and hepatic stellate cell (HSC) remain unclear. In this research, as a result of screening of amino acids using liver fibrosis induced by DMN administration, L-cysteine was selected as a suppressor of liver fibrosis. Furthermore, the number of activated HSCs, which increased in the fibrotic liver after DMN administration, was decreased in L-cysteine-fed rats. Treatment of freshly isolated HSCs with L-cysteine resulted in inhibition of the increase in smooth muscle alpha-actin (alphaSMA) expression by HSCs and BrdU incorporation into the activated HSCs. These findings suggest that L-cysteine is effective against liver fibrosis. The mechanism of inhibition of fibrosis in the liver is surmized to be direct inhibition of activated HSC proliferation and HSC transformation by L-cysteine. PMID- 12732203 TI - Prolonged gene expression in primary porcine pancreatic cells using an Epstein Barr virus-based episomal vector. AB - Epstein-Barr virus (EBV)-based plasmids containing the origin of replication (oriP) and EBV nuclear antigen 1 (EBNA-1) are well known for the stable episomal maintenance in human cells. In order to clarify whether an EBV-based plasmid can be maintained stably in the porcine pancreatic cells which are the primary candidate sources of islet xenotransplantation, we constructed pEBVGFP encoding the green fluorescent protein (GFP). Monolayer culture of the porcine neonatal pancreatic cells was lipofected with pEBVGFP or pGFP which was derived from pEBVGFP by deleting out oriP and EBNA-1. pEBVGFP significantly prolonged GFP expression not only in human cell lines but also in the primary porcine pancreatic cells compared with pGFP. Interestingly, the duct cells that are believed as the pancreatic precursor cells were preferentially transfected and conveniently enriched among the mixed primary cell populations using a hygromycin B selection. To our knowledge, this is the first report suggesting the potential application of an EBV-based plasmid for the extended gene expression in the primary porcine pancreatic duct cells. PMID- 12732202 TI - Regulatory role of diacylglycerol kinase gamma in macrophage differentiation of leukemia cells. AB - Although nine diacylglycerol kinase (DGK) isozymes have been identified, our knowledge of their individual functions is still limited. Here we report that the levels of DGKgamma mRNA/protein in human leukemia HL-60 and U937 cells were rapidly and markedly decreased upon cellular differentiation into macrophages. In contrast, the enzyme expression remained almost unchanged in granulocytic differentiation pathway. Interestingly, the overexpression of wild-type or constitutively active DGKgamma, but not its kinase-dead mutant, markedly inhibited phorbol ester-induced cell attachment and nonspecific esterase activity, which are hallmarks of macrophage differentiation. We noted in this case that no effects were observed for the corresponding constructs of a closely related isozyme, DGKalpha. Prior to the cell attachment, phorbol ester induced translocation of DGKgamma from the cytoplasm to the cell periphery, resulting in its co-localization with F-actin together with protein kinase Cdelta. The results suggest that DGKgamma negatively regulates macrophage differentiation through its catalytic action operating on the cytoskeleton. PMID- 12732204 TI - Fabrication and photoelectric response of poly(allylamine hydrochloride)/PM thin films by layer-by-layer deposition technique. AB - Thin films of poly(allylamine hydrochloride) (PAH) and bacteriorhodopsin (bR) embedded in purple membrane (PM) have been prepared by layer-by-layer (LBL) self assembly technique. The results obtained by UV-Vis spectroscopy and atomic force microscopy (AFM) analysis showed that the biological activity of bR was preserved and PM fragments could be well oriented onto the ITO substrate. A photo electrochemical cell with the structure of ITO/(PAH/PM)(n)/electrolyte (0.5M KCl)/Pt was fabricated and studied. The photocurrent peaks of (PAH/PM)(6) corresponding to light-on and light-off were about 200 and 100 nA/cm(2), respectively, with the former enhanced 30% higher than that of the reference films made of (PDAC/PM)(6). PMID- 12732205 TI - High glucose accelerates MCP-1 production via p38 MAPK in vascular endothelial cells. AB - In diabetes mellitus (DM), hyperglycemia causes cardiovascular lesions through endothelial dysfunction. Monocyte chemoattractant protein-1 (MCP-1) is implicated in the pathogenesis of cardiovascular lesions. By using human umbilical vein endothelial cells, we investigated the effect of hyperglycemia on MCP-1 production and its signaling pathways. Chronic incubation with high glucose increased mRNA expression and production rate of MCP-1 in a time (1-7 days)- and concentration (10-35 mM)-dependent manner. Chronic exposure to high glucose resulted in enhancement of generation of reactive oxygen species (ROS), as determined by increasing level of 2,7-dichlorofluorescein (DCF), and subsequent activation of p38 mitogen-activated protein kinase (MAPK). Neither c-Jun NH(2) terminal kinase nor extracellular signal-regulated kinase1/2 was affected. SB203580 or FR167653, p38 MAPK specific inhibitors, completely suppressed MCP-1 expression. Catalase suppressed p38 MAPK phosphorylation and MCP-1 expression. These results indicate that hyperglycemia can accelerate MCP-1 production through the mechanism involving p38 MAPK, ROS-sensitive signaling pathway, in vascular endothelial cells. PMID- 12732206 TI - ULBP4 is a novel ligand for human NKG2D. AB - The ULBPs are a family of MHC class I-related molecules. We have previously shown that ULBPs 1, 2, and 3 are functional ligands of the NKG2D/DAP10 receptor complex on human natural killer (NK) cells. Here, we describe a new member of the ULBP family, ULBP4, which contains predicted transmembrane and cytoplasmic domains, unlike the other ULBPs, which are GPI-linked proteins. Transduction of ULBP4 into EL4 cells confers the ability to bind recombinant NKG2D and mediates increased cytotoxic activity by human NK cells, consistent with the role of ULBPs as ligands for the NKG2D/DAP10 activating receptors. Tissue expression of ULBP4 differs from other members of the family, in that it is expressed predominantly in the skin. PMID- 12732207 TI - Inflammatory cytokines upregulate nephrin expression in human embryonic kidney epithelial cells and podocytes. AB - Nephrin is an important regulator of the glomerular filtration barrier and its malfunction is associated with severe proteinuria. In this study we show that exposure of human embryonic kidney epithelial A293 cells to the proinflammatory cytokine interleukin-1beta (IL-1beta) causes a dose-dependent upregulation of nephrin mRNA level. Time-course analyses reveal first significant increases in nephrin mRNA levels after 4h of stimulation. Furthermore, nephrin protein is also elevated by IL-1beta treatment. Tumor necrosis factor-alpha (TNFalpha) exerted a comparable effect on nephrin mRNA and protein expression. The IL-1beta-induced upregulation of nephrin expression occurs independently of nitric oxide (NO) generation, since the NO-synthase inhibitor N(G)-monomethyl-L-arginine does not block the IL-1beta effect. Mechanistically, we found that the IL-1beta-induced response does not involve protein kinase C, protein kinase A, the classical mitogen-activated protein kinase (MAPK), the stress-activated p38-MAPK, or the NF kappaB cascade, since selective inhibitors of these pathways were unable to alter the IL-1beta response. Moreover, neither unselective cyclooxygenase (COX) inhibitors, like indomethacin, nor COX-2-selective inhibitors, like flosulide and NS 398, nor the anti-inflammatory glucocorticoid dexamethasone were able to alter IL-1beta-induced nephrin expression. The only inhibitor that was able to block IL 1beta- and TNFalpha-induced nephrin upregulation was rottlerin, which has been suggested to act as a selective PKCdelta inhibitor. However, concerning cytokine triggered nephrin expression, rottlerin action involved inhibition of another still to be identified protein kinase. Importantly, cytokine-induced upregulation of nephrin expression was also confirmed in primary human podocytes. In summary, these data show for the first time that inflammatory cytokines like IL-1beta or TNFalpha can upregulate nephrin expression and this mechanistically involves a rottlerin-sensitive protein kinase. PMID- 12732209 TI - Telomerase upregulates expression levels of interleukin (IL)-1alpha, IL-1beta, IL 6, IL-8, and granulocyte-macrophage colony-stimulating factor in normal human fibroblasts. AB - Expression of human telomerase reverse transcriptase (hTERT) in normal human fibroblast cell strain, TIG-3, extends their replicative life span. We found that expression levels of interleukin (IL)-1alpha, IL-1beta, IL-6, IL-8, and granulocyte-macrophage colony-stimulating factor (GM-CSF) mRNA were up-regulated in hTERT-induced fibroblasts irrespective of population doubling level (PDL). Expression levels of these cytokines were low in growing young TIG-3 cells and in control vector-transfected TIG-3 cells but were up-regulated in growth-arrested young cells maintained at high cell density. In senescent TIG-3 cells, expression of IL-1beta, IL-6, and GM-CSF was moderately increased. These results indicate that the introduction of hTERT into normal fibroblasts up-regulates the expression of some inflammatory cytokines, and caution should be paid when introducing the hTERT gene to establish cell lines with normal phenotype. PMID- 12732208 TI - Increase in putrescine, amine oxidase, and acrolein in plasma of renal failure patients. AB - Since polyamines have been suggested to be one of the uremic "toxins," the levels of each polyamine, its oxidized product, acrolein, and amine oxidase in plasma of patients with renal failure were investigated. The level of putrescine was increased, whereas the level of spermine was decreased in the plasma of patients with renal failure. The patients also had increased serum amine oxidase activity leading to increased degradation of spermine. Both levels of free and protein conjugated acrolein were also increased in plasma of patients with renal failure. The accumulated acrolein found as protein conjugates was equivalent to 180 microM, which was 6-fold higher than in plasma of normal subjects. It was found that acrolein is mainly produced by polyamine oxidase in plasma. A cell lysate containing polyamine oxidase was cytotoxic in the presence of spermine. Our results indicate that the level of acrolein is well correlated with the degree of seriousness of chronic renal failure. PMID- 12732210 TI - PREP1, MEIS1 homolog protein, regulates PF4 gene expression. AB - We have previously demonstrated that homeodomain proteins, MEIS1 and PBXs, transactivate the PF4 gene through the novel regulatory element termed TME. This study focuses on Pbx regulating protein 1 (PREP1), a MEIS1 homolog protein, for its transcriptional activity in the PF4 promoter. PREP1 binds to the TME in HEL cells. PREP1 was expressed in human megakaryocytes that differentiated from CD34(+) cells. EMSA shows that either PREP1 by itself or PREP1/PBX complexes bind to the two TGACAG motifs in the TME and activate the PF4 promoter. Furthermore, PREP1 and PREP1/PBX complexes synergistically activate the PF4 promoter with GATA 1 and ETS-1. These data demonstrate that PREP1 is also an important transcription factor that regulates PF4 gene expression such as MEIS1. Additionally, these data imply functional similarities and differences between PREP1 and MEIS1 in the regulation of PF4 gene expression. PMID- 12732211 TI - Potentiation of invasive activity of hepatoma cells by reactive oxygen species is mediated by autocrine/paracrine loop of hepatocyte growth factor. AB - We have already reported that reactive oxygen species (ROS) promote rat ascites hepatoma cell invasion beneath mesentery-derived mesothelial cell monolayer. To investigate the mechanism for this, we examined the involvement of motility factors, particularly hepatocyte growth factor (HGF). Rat ascites hepatoma cell line of AH109A expressed HGF and c-Met mRNAs. Treatment with ROS augmented amounts of HGF mRNA in AH109A and HGF concentration in the medium. ROS also induced HGF gene expression in mesothelial cells. Exogenously added HGF enhanced invasive activity of AH109A cells, but exerted no effect on proliferation. AH109A cells pretreated with ROS showed an increased invasive activity, which was cancelled by simultaneous pretreatment with anti-HGF antibody. These results suggest that the invasive activity of AH109A is mediated by the autocrine and paracrine pathways of HGF, and ROS potentiate invasive activity by inducing gene expression of HGF in AH109A and mesothelial cells. PMID- 12732212 TI - PAR-3 is a low-affinity substrate, high affinity effector of thrombin. AB - A polypeptide corresponding to the extracellular domain of protease-activated receptor 3 (PAR-3) is hydrolyzed by thrombin slowly because of high K(M) (>100 microM). However, thrombin is found to bind two PAR-3, one without catalyzing hydrolysis or blocking the active site, while the other is hydrolyzed. In a solvent lacking Na(+), hydrolysis of a nitroanilide substrate is enhanced 1.6 fold by addition of PAR-3 polypeptide, with half-saturation at 2.5 microM. In contrast, the fibrinogen clotting activity of thrombin is inhibited completely by PAR-3, with a K(I) of 3 microM. None of the activities of thrombin are affected by addition of 50 microM PAR-4 polypeptide. Thus, PAR-3 in low concentrations binds thrombin in a configuration that blocks the anion-binding exosite but not the catalytic site, while hydrolysis of PAR-3, PAR-4, and other substrates that do not interact with exosite I persists. The allosteric effect of PAR-3 is characteristic of that of Na(+). PMID- 12732213 TI - Ghrelin gene in cichlid fish is modulated by sex and development. AB - We have identified the complementary and genomic DNA sequences of the novel growth hormone-releasing peptide, ghrelin, in the cichlid fish (tilapia Oreochromis niloticus). The tilapia ghrelin precursor cDNA was 855 bp long, consisting of 119 bp of 5(')-untranslated region, 324 bp open reading frame, and 412 bp of 3(')-untranslated region. The serine residues at positions 2 and 3 of the "active core" (GSSF) of the ghrelin mature peptide are conserved between tilapia and the mammalian species. The tilapia ghrelin gene has four exons and three introns and resembles the structure of other known ghrelin genes. RT-PCR analysis revealed ghrelin mRNA predominantly expressed in the stomach but absent in the brain, pituitary, heart, kidney, ovary, and testis. Real-time PCR analysis showed an age/body size dependent increase in gastric ghrelin, which stagnated at 7 cm body size (onset of maturation). Ghrelin mRNA levels were unchanged in food deprived sexually mature animals but were significantly higher in females compared to males. The present study shows that the structure of ghrelin peptide is highly conserved, and the reported differences in somatic and gonadal growth in tilapia could be a consequence of age- and sex-related synthesis of gastric ghrelin. PMID- 12732215 TI - Cloning and characterization of the 5(')-flanking region of the human ghrelin gene. AB - Ghrelin, a novel growth hormone releasing peptide, was recently isolated from stomach. We have cloned and characterized the 5(')-flanking region, containing from -2000 to -1 upstream from the translation start site of the human ghrelin gene. There was neither typical GC nor CAAT box but there were a TATATAA element and putative binding sites for several transcription factors. Ghrelin promoter was activated only in human stomach derived ECC10 cells among several cell lines examined. Functional analysis showed that promoter activity was increased by deletion of nucleotides from -2000 to -605 whereas it was decreased by further deletion and that the TATATAA element is not functioning. Glucagon and its second messenger cAMP enhanced the promoter activity, suggesting that stimulated transcription of ghrelin gene by glucagon might be responsible for increased ghrelin production during fasting at least in part. These initial characterizations will facilitate further studies of the regulatory mechanisms for ghrelin gene expression. PMID- 12732214 TI - Sodium orthovanadate potentiates EGCG-induced apoptosis that is dependent on the ERK pathway. AB - Epigallocatechin-3-gallate (EGCG) is a potent chemopreventive agent in many test systems and has been shown to inhibit tumor promotion and induce apoptosis. In the present study, we determined the effect of vanadate, a potent inhibitor of tyrosine phosphatase, on EGCG-induced apoptosis. Investigation of the mechanism of EGCG or vanadate-induced apoptosis revealed induction of caspase 3 activity and cleavage of phospholipase-gamma1 (PLC-gamma1). Furthermore, vanadate potentiated EGCG-induced apoptosis by mitogen-activated protein kinase (MAPK) signaling pathway. Treatment with EGCG plus vanadate for 24h produced morphological features of apoptosis and DNA fragmentation in U937 cells. This was associated with cytochrome c release, caspase 3 activation, and PLC-gamma1 degradation. EGCG plus vanadate activates multiple signal transduction pathways involved in coordinating cellular responses to stress. We demonstrate a requirement for extracellular signal-regulated protein kinase (ERK), a member of the mitogen-activated protein kinase family in EGCG plus vanadate-induced apoptosis in U937 cells. Elevated ERK activity that contributed to apoptosis by EGCG plus vanadate was supported by PD98059 and U0126, chemical inhibitor of MEK/ERK signaling pathway, prevented apoptosis. Taken together, our finding suggests that ERK activation plays an active role in mediating EGCG plus vanadate induced apoptosis of U937 cells and functions upstream of caspase activation to initiate the apoptotic signal. PMID- 12732216 TI - Differentiation-dependent up-regulation of p47(phox) gene transcription is associated with changes in PU.1 phosphorylation and increased binding affinity. AB - The p47(phox) gene encodes a cytosolic component of the phagocytic NADPH oxidase complex. Expression of p47(phox) is both tissue-specific and developmentally regulated. Stable transfection of the myeloid cell lines PLB985 and HL60, with reporter gene constructs containing as little as 58 bp of proximal promoter sequence, was capable of directing significant reporter gene activity in myeloid cells, which increased significantly on induction of myeloid differentiation. EMSA analysis of a binding site for the Ets family member, PU.1, located at positions -39 to -44 revealed that the pattern of complex formation changed significantly on induction of myeloid differentiation. All EMSA complexes were competed by a functional PU.1 binding site and could be supershifted in the presence of polyclonal anti-PU.1 antibody. Reaction of EMSA complexes with anti phosphoserine antibody, treatment with phosphatase, or Western blotting of proteins captured on the PU.1 binding site, was used to demonstrate that the changes in PU.1 complex formation dependent on myeloid differentiation were associated with increased levels of PU.1 phosphorylation. Furthermore, the more highly phosphorylated forms of PU.1 were shown to have a greater affinity for the p47(phox) PU.1 consensus binding site. Up-regulated transcriptional activity in response to myeloid differentiation can therefore be correlated with increased levels of PU.1 phosphorylation and a greater binding affinity. PMID- 12732217 TI - Lactate-sensitive response elements in genes involved in hyaluronan catabolism. AB - Tissue anoxia occurs early in wound healing. This is accompanied by production of lactate followed by increased hyaluronan and CD44 expression, suggesting a cause and effect relationship. Fibroblasts increased hyaluronan and CD44 when lactate was added to cultures. Increased deposition of hyaluronan correlates with greater turnover. In current models of hyaluronan catabolism, it is tethered to cell surfaces by CD44 in caveolin-enriched invaginations. It is cleaved to 20-kDa fragments by Hyal-2 on the plasma membrane, endocytosed, and delivered ultimately to lysosomes, and further digested by Hyal-1. Sequence analyses of promoter regions of genes for CD44, caveolin-1, Hyal-1, and -2 revealed multiple AP-1 and ets-1 response elements. To test their relevance, RNA from lactate-treated fibroblasts was analyzed by reverse transcriptase-polymerase chain reaction. Increased transcripts of c-fos, c-jun, c-ets, Hyal-1, -2, CD44, and caveolin-1 mRNAs were observed. We have thus identified lactate-activated genes important in the wound healing responses. Similar responses facilitating tumor progression, the Warburg effect, may share such mechanisms. PMID- 12732219 TI - Trajectories of brain development: point of vulnerability or window of opportunity? AB - Brain development is a remarkable process. Progenitor cells are born, differentiate, and migrate to their final locations. Axons and dendrites branch and form important synaptic connections that set the stage for encoding information potentially for the rest of life. In the mammalian brain, synapses and receptors within most regions are overproduced and eliminated by as much as 50% during two phases of life: immediately before birth and during the transitions from childhood, adolescence, to adulthood. This process results in different critical and sensitive periods of brain development. Since Hebb (1949) first postulated that the strengthening of synaptic elements occurs through functional validation, researchers have applied this approach to understanding the sculpting of the immature brain. In this manner, the brain becomes wired to match the needs of the environment. Extensions of this hypothesis posit that exposure to both positive and negative elements before adolescence can imprint on the final adult topography in a manner that differs from exposure to the same elements after adolescence. This review endeavors to provide an overview of key components of mammalian brain development while simultaneously providing a framework for how perturbations during these changes uniquely impinge on the final outcome. PMID- 12732220 TI - Risk-taking behavior in adolescent mice: psychobiological determinants and early epigenetic influence. AB - Epidemiological research has emphasized that adolescence is associated with some temperamental and behavioral traits that are typical of this age and that might substantially contribute to both psychological and psychobiological vulnerability. The contribution of the important developmental rearrangements in neurobiological and neuroendocrinological processes has received surprisingly little investigation. The present review summarizes recent work in animal models, indicating that adolescent rodents exhibit marked peculiarities in their spontaneous behavioral repertoire. When compared to adults, adolescents show an unbalanced and 'extremes-oriented' behavior, consisting of an increased novelty seeking, together with decreased novelty-induced stress and anxiety, an increased risk-taking behavior in the plus-maze, as well as elevated levels of impulsivity and restlessness. Age-related discontinuities in the function of monoaminergic systems, which are a main target of abused drugs, can perhaps account for such a profile. In particular, a peculiar function within reward-related dopaminergic brain pathways actually seems to underlie the search for novel and rewarding sensations, as well as changes in the magnitude of psychostimulant effects. The role played by early epigenetic factors in the shaping of novelty-seeking behavior of adolescent and adult rodents are also reviewed. Two examples are considered, namely, subtle variations in the hormonal milieu as a function of intrauterine position and precocious or delayed maturation of nutritional independence as a function of changes in time of weaning. As for spontaneous drug consumption, a prominent vulnerability to the oral intake of nicotine during early adolescence is reported. In conclusion, adolescence in rodents may represent a suitable animal model with enough face- and construct-validity. Actually, this model is able to show behavioral features that resemble those found in human adolescents, including vulnerability to the consumption of psychoactive drugs. PMID- 12732221 TI - The neurobiological consequences of early stress and childhood maltreatment. AB - Early severe stress and maltreatment produces a cascade of neurobiological events that have the potential to cause enduring changes in brain development. These changes occur on multiple levels, from neurohumoral (especially the hypothalamic pituitary-adrenal [HPA] axis) to structural and functional. The major structural consequences of early stress include reduced size of the mid-portions of the corpus callosum and attenuated development of the left neocortex, hippocampus, and amygdala. Major functional consequences include increased electrical irritability in limbic structures and reduced functional activity of the cerebellar vermis. There are also gender differences in vulnerability and functional consequences. The neurobiological sequelae of early stress and maltreatment may play a significant role in the emergence of psychiatric disorders during development. PMID- 12732222 TI - Early experience as a determinant of adult behavioural responses to reward: the effects of repeated maternal separation in the rat. AB - Depression is a major public health concern, representing one of the most significant causes of disability and morbidity. Despite significant advances in the definition of specific cognitive, emotional and neural dysfunctions that are associated with depression, there has been frustratingly little progress in the elucidation of plausible aetiological and pathophysiological mechanisms. The complex, multi-system dysfunctions of depressive illness do not lend themselves to hypothesis-driven, systematic manipulation in patients. For this reason, there is a need to develop valid and reliable models of affective psychopathology in laboratory animals. In this paper, we review briefly some of our previous work demonstrating that a specific periodic neonatal maternal separation procedure leads to a robust constellation of behavioural changes in the adult rat that resemble core aspects of human depressive psychopathology. We also present data from a study of the adult effects of the same manipulation on electrical intracranial self-stimulation behaviour. These data further support the hypothesis that it is possible to model vulnerability to anhedonia in the adult rat by manipulation of early experience. PMID- 12732223 TI - Long-term neurobehavioural impact of the postnatal environment in rats: manipulations, effects and mediating mechanisms. AB - The major characteristics of the postnatal environment of the rat pup are its mother and littermates. The pup, which is poorly developed at birth, matures rapidly in this environment, and regulates the behaviour and physiology of the dam and littermates, as well as vice versa. The study of the impact of the rat's postnatal environment on its long-term neurobehavioural development is of fundamental importance. In fact, it is one of the major examples--at the interface of the biological, social and medical sciences--of animal models for the study of the interaction between the environment and the genome in both the acute and chronic regulation of the phenotype. Specific experimental manipulations of the rat postnatal environment have been demonstrated to exert robust and marked effects on neurobiological, physiological and behavioural phenotypes in adulthood. In the present review we present some of the major findings, including some original data, and discuss what these existing data can tell us about the long-term neurobehavioural effects of the postnatal environment in rats, the external and internal mechanisms that mediate these effects, and the most appropriate directions for future basic and applied research in this area. PMID- 12732224 TI - Early disruption of the mother-infant relationship: effects on brain plasticity and implications for psychopathology. AB - Early environmental manipulations can impact on the developing nervous system, contributing to shape individual differences in physiological and behavioral responses to environmental challenges. In particular, it has been shown that disruptions in the mother-infant relationship result in neuroendocrine, neurochemical and behavioural changes in the adult organism, although the basic mechanisms underlying such changes have not been completely elucidated. Recent data suggest that neurotrophins might be among the mediators capable of transducing the effects of external manipulations on brain development. Nerve growth factor and brain-derived neurotrophic factor are known to play a major role during brain development, while in the adult animal they are mainly responsible for the maintenance of neuronal function and structural integrity. Changes in the levels of neurotrophic factors during critical developmental stages might result in long-term changes in neuronal plasticity and lead to increased vulnerability to aging and to psychopathology. PMID- 12732225 TI - The long-term psychobiological consequences of intermittent postnatal separation in the squirrel monkey. AB - Long-term effects of early experiences have been well documented in rodents, non human primates and humans. The influence of early experiences on the regulation of the hypothalamic-pituitary-adrenal has demonstrated that this biological system is highly plastic and is permanently modified by early experiences. The effects of brief periods of maternal separation in rodents tend to result in a reduction of the stress response, whereas longer periods of maternal separation produce an animal that is hyperresponsive. The effect of disruptions of mother infant interactions on the HPA axis in non-human primates has not been clearly established. However, the primate research has primarily focused on more permanent separation paradigms and thus it is difficult to compare the rodent studies with those conducted in primates. The present study attempted to use a separation procedure in monkeys that more closely resembled the rodent model. Intermittent separation during infancy followed by reunion with the mother were shown to lead to a significant reduction in both the cortisol response and the response of the noradrenergic system following social isolation in juvenile squirrel monkeys at 2 and 3 years of age. The behavioral response (vocalizations) was also reduced. PMID- 12732226 TI - Birth insult interacts with stress at adulthood to alter dopaminergic function in animal models: possible implications for schizophrenia and other disorders. AB - Altered subcortical dopaminergic activity is thought to be involved in the pathophysiology of several disorders including schizophrenia, substance abuse and attention deficit hyperactivity disorder. Epidemiological studies have implicated perinatal insults, particularly obstetric complications involving fetal or neonatal hypoxia, as etiological risk factors for schizophrenia. This suggests the possibility that perinatal hypoxia might have lasting effects on dopaminergic function. In animal models, dopaminergic systems appears to be particularly vulnerable to a wide range of perinatal insults, resulting in persistent alterations in function of mesolimbic and mesostriatal pathways. This review summarizes recent work characterizing long-term changes in dopaminergic function and biochemistry in models of Caesarean section (C-section) birth and of C section birth with added global anoxia in the rat and guinea pig. C-section birth and C-section with anoxia appear to be two distinct hypoxic birth insults, with somewhat differing patterns of lasting effects on dopamine systems. In addition, birth insult alters the manner in which dopaminergic function is regulated by stress at adulthood. The possible relevance of these finding to effects of human birth procedures is discussed. PMID- 12732227 TI - Sex differences in brain maturation in maltreatment-related pediatric posttraumatic stress disorder. AB - BACKGROUND: Recent investigations suggested that pediatric posttraumatic stress disorder (PTSD) is associated with adverse brain development. However, sex differences are poorly understood. METHODS: In this study, 61 medically healthy children and adolescents (31 males and 30 females) with chronic PTSD secondary to abuse, who had similar trauma and mental health histories, and 122 healthy controls (62 males and 60 females) underwent comprehensive psychiatric assessments and an anatomical MRI brain scan. RESULTS: When gender groups were analyzed separately, findings of larger prefrontal lobe CSF volumes and smaller midsagittal area of the corpus callosum subregion 7 (splenium) were seen in both boys and girls with maltreatment-related PTSD compared to their gender-matched comparison subjects. Subjects with PTSD did not show the normal age related increases in the area of the total corpus callosum and its region 7 (splenium) compared to non-maltreated subjects; however, this finding was more prominent in males with PTSD. Significant sex by group effects demonstrated smaller cerebral volumes and corpus callosum regions 1 (rostrum) and 6 (isthmus) in PTSD males and greater lateral ventricular volume increases in maltreated males with PTSD than maltreated females with PTSD. CONCLUSIONS: These data suggest that there are sex differences in the brain maturation of boys and girls with maltreatment-related PTSD. Longitudinal MRI brain investigations of childhood PTSD and the relationship of gender to psychosocial outcomes are warranted. PMID- 12732228 TI - Prenatal stress and long-term consequences: implications of glucocorticoid hormones. AB - We have shown that prenatal restraint stress (PNRS) induces higher levels of anxiety, greater vulnerability to drugs, a phase advance in the circadian rhythm of locomotor activity and an increase in the paradoxical sleep in adult rats. These behavioral effects result from permanent modifications to the functioning of the brain, particularly in the feedback mechanisms of the hypothalamic pituitary-adrenal (HPA) axis: the secretion of corticosterone is prolonged after stress and the number of the central glucocorticoid receptors is reduced. These abnormalities are associated with modifications in the synthesis and/or release of certain neurotransmitters. Dysfunction of the HPA axis is due, in part, to stress-induced maternal increase of glucocorticoids, which influences fetal brain development. Some biological abnormalities in depression can be related to those found in PNRS rats reinforcing the idea of the usefulness of PNRS rats as an appropriate animal model to study new pharmacological approaches. PMID- 12732229 TI - Prenatal elevation of endocannabinoids corrects the unbalance between dopamine systems and reduces activity in the Naples High Excitability rats. AB - Several evidences suggest that endocannabinoids exert a neurotrophic effect on developing mesencephalic dopamine neurons. Since an altered mesocorticolimbic system seems to underlie hyperactivity and attention deficit in clinical and animal studies of attention deficit hyperactivity disorder (ADHD), prenatal elevation of anandamide has been induced in Naples high excitability (NHE) rats by inhibition of its reuptake. To this aim, pregnant NHE and random-bred females received a subcutaneous injection of AM-404 (1 mg/kg) or vehicle daily from E11 until E20. Young adult male offsprings were exposed to a spatial novelty (Lat maze) for 30 min and the behavior was videotaped and analysed for indices of activity (travelled distance, rearing frequency) and attention (rearing duration). Moreover, morphological analysis of the brains was carried out that pertained to cytochrome oxydase as marker of metabolic activity and thyrosine hydroxylase as marker of the dopamine systems. The results indicate that prenatal AM-404 treatment significantly reduces activity by about 20% during the entire testing period and modifies the distribution of scanning times towards short duration episodes in the first part of the test only in NHE-treated rats. In addition, image analysis revealed a significant increase in relative optical density of TH+terminals in the dorsal striatum and substantia nigra of AM-404 treated NHE rats and minor changes in the dorsal cortex of AM-404 treated NRB rats. The data suggest a corrected unbalance between the two dopamine systems that apparently leads to reduced hyperactivity and modified scanning times in this animal model of ADHD. This, in turn, might open new strategies in the treatment of a subset of ADHD cases. PMID- 12732230 TI - Animal models of mental retardation: from gene to cognitive function. AB - About 2-3% of all children are affected by mental retardation, and genetic conditions rank among the leading causes of mental retardation. Alterations in the information encoded by genes that regulate critical steps of brain development can disrupt the normal course of development, and have profound consequences on mental processes. Genetically modified mouse models have helped to elucidate the contribution of specific gene alterations and gene-environment interactions to the phenotype of several forms of mental retardation. Mouse models of several neurodevelopmental pathologies, such as Down and Rett syndromes and X-linked forms of mental retardation, have been developed. Because behavior is the ultimate output of brain, behavioral phenotyping of these models provides functional information that may not be detectable using molecular, cellular or histological evaluations. In particular, the study of ontogeny of behavior is recommended in mouse models of disorders having a developmental onset. Identifying the role of specific genes in neuropathologies provides a framework in which to understand key stages of human brain development, and provides a target for potential therapeutic intervention. PMID- 12732231 TI - Age-associated sex differences in response to food deprivation in two animal tests of anxiety. AB - The effects of mild food deprivation (7 days of food restricted to once daily feeding to maintain body weights at 85% of free-feeding weights) were examined in adult male and female and adolescent female rats tested in the elevated plus maze and social interaction tests of anxiety. In adult male rats, food deprivation appeared to have an anxiolytic effect in the plus-maze as it significantly increased the percentage of entries onto open arms and the percentage of time spent on the open arms, without changing the number of closed arm entries. There were no effects of food deprivation in adult females, although in adolescent females food deprivation significantly increased the percentage of open arm entries rats. Adolescent female rats have female brains, but do not have circulating gonadal hormones and thus these results suggest that circulating female gonadal hormones are able to suppress some of the effects of mild food deprivation in the plus-maze. In the social interaction test, there were no effects of food deprivation in any group on the time spent in social interaction. There were opposite effects on locomotor activity in the adult male and female rats, with deprivation increasing activity in males and decreasing it in females. There were no effects of food deprivation on locomotor activity in the adolescent females, suggesting that circulating gonadal hormones were responsible for the bidirectional effects in the adult rats. In both tests there were age-associated differences in the female rats, with the adolescent females being less anxious (higher percentage of open arm entries and increased social interaction) than the adults. PMID- 12732232 TI - Ontogenesis of behavioral sensitization and conditioned place preference induced by psychostimulants in laboratory rodents. AB - The present review deals mainly with the ontogenesis of two important phenomena involved in vulnerability to several neuropsychiatric disorders, namely with drug induced sensitization (both contextual and non-contextual) and with conditioned place preference. The term 'infancy' covers the first three postnatal weeks during development in rats and mice. Conversely, the term 'adolescence' may cover the whole postnatal period ranging from weaning (PND 21) to adulthood (at least PND 60) or specifically the period around the onset of puberty (animals aged 33 44 days). Recent studies in rats demonstrated that the establishment of a context dependent sensitization appears during the first (for repeated drug administration) or during the second (for a single drug administration) postnatal week. However, the memory of drug-context association is transient in developing pups (lasting one or two days following the drug pretreatment). The long-term retention of drug-context associations matures progressively, and is complete by the third week of postnatal life. Finally, those mechanisms responsible for an adult-like profile of context-independent pharmacological sensitization appear later during ontogenesis, being mature by the fourth week of postnatal life. Another set of experiments extended this ontogenetic characterization by comparing adolescent and adult mice. When compared to the latter, the former subjects exhibit a greater amphetamine-induced locomotor sensitization, almost no sensitization of aversive stereotyped behaviors, and a less marked place conditioning. The strength of the drug-induced place conditioning was also directly compared with the unconditioned novelty-seeking drive. In conclusion, neonatal rats are able to show a relatively short-lasting retention of sensitized drug effects (short-term sensitization), whereas the ability to exhibit relatively long-lasting sensitized effects matures progressively during infancy (long-term sensitization). On the other hand, adolescent mice show a reduced sensitization of drug-induced psychotic symptoms, together with a more marked sensitization of arousing and euphorigenic properties of the drug and a reduced incentive memory of its hedonic effects. These age-related changes do imply very different degrees of vulnerability to drug addiction and several other neuropsychiatric disorders. PMID- 12732233 TI - Cardiac autonomic reactivity and salivary cortisol in men and women exposed to social stressors: relationship with individual ethological profile. AB - The degree of cardiovascular stress responsivity and its possible implications for the onset and progression of cardiovascular pathologies seem to be linked to the individual strategy of behavioral coping with stressors. This study was designed to investigate the relationship among cardiac autonomic, endocrine and behavioral responses to real-life stress episodes. Thirty university students were exposed to two brief social challenges (stress interviews), during which the state of sympathovagal balance (time-domain indexes of heart rate variability) and a number of non-verbal behaviors were quantified. Psychometric measurements were also obtained via SPRAS questionnaire, administered just after each stress interview. Samples of saliva were collected for cortisol determination immediately prior and after the experimental session. Subjects showing higher levels of sympathetic dominance were characterized by higher scores of submissive behavior, larger cortisol increments, and higher perception of psychophysiological arousal. A clear consistency in the individual response to the two stress interviews was found, at the behavioral, physiological and psychophysiological level. Finally, the gender of the subjects did not clearly influence their stress responsivity. These results support the hypothesis of a close relationship between the degree of physiological arousal and the style of behavioral adaptation to social stressors. PMID- 12732234 TI - A secretin i.v. infusion activates gene expression in the central amygdala of rats. AB - For the last 100 years secretin has been extensively studied for its hormonal effects on digestion. Recent observations that the deficits in social reciprocity skills seen in young (3-4-year-old) autistic children are improved after secretin infusions suggest an additional influence on neuronal activity. We show here that i.v. administration of secretin in rats induces Fos protein expression in the neurons of the central amygdala as well as the area postrema, bed nucleus of the stria terminalis, external lateral parabrachial nucleus and supraoptic nucleus. However, secretin infusion did not induce Fos expression in the solitary tract nucleus or paraventricular nucleus, regions normally activated by related peptides such as cholecystokinin. The peak blood levels of secretin that induce Fos protein expression in rat brain are similar to the peak blood levels observed during i.v. treatment with secretin in humans. The amygdala is known to be critical for developing reciprocal social interaction skills and abnormalities in this brain region have been demonstrated in autistic children. PMID- 12732235 TI - Subcellular localization of the voltage-dependent potassium channel Kv3.1b in postnatal and adult rat medial nucleus of the trapezoid body. AB - A pre-embedding immunocytochemical method was used to study the subcellular distribution of the voltage-dependent potassium channel Kv3.1b in the medial nucleus of the trapezoid body (MNTB) in developing and adult rat. The main finding was the localization of the channel in specific membrane compartments of the calyces of Held and principal globular neurons. Thus, at postnatal day (P) 9 immunoparticles were densely localized in plasma membranes of globular cell bodies and their main dendrites. At P16, a strong Kv3.1b labeling was still observed in these globular cell compartments, but the most remarkable feature was the presence of immunoparticles in synaptic terminal membranes of the calyces of Held. However, the presynaptic and postsynaptic specializations of the calyx of Held-globular cell synapses were virtually devoid of immunoparticles. This same subcellular distribution of Kv3.1b was seen in adult, with membranes of calycine terminals more uniformly labeled. The developmental profile of Kv3.1b expression in MNTB coincides with the functional maturation of the calyx of Held-principal globular neuron synapse. The presence of the channel in this system is crucial for the high-frequency synaptic transmission of auditory signals. PMID- 12732236 TI - Optical detection of dendritic spike initiation in hippocampal CA1 pyramidal neurons. AB - Previous studies have shown that spikes can be generated in the dendrites of CA1 pyramidal neurons. Some have suggested that, in response to synaptic inputs, spikes are initiated near the soma and propagate back into the dendrites, but some recent studies have shown that intense synaptic inputs initiate spikes in the dendrite. Here, we report the optical detection of spike propagation along the apical dendrites of hippocampal pyramidal neurons. Rat hippocampal slices were stained with the fluorescent voltage-sensitive dye, JPW1114, and optical signals monitored using a 16 x 16 photodiode array system at a frame rate of 4 kHz. A stimulating electrode was placed at the boundary between the stratum (str.) lacnosum-moleculare and the str. radiatum to stimulate the Schaffer collateral, and fast and slow signal components were detected in the dendritic and somatic regions. By comparing the optical signals with whole-cell recordings, we confirmed that the fast component was due to a population of dendritic spikes in pyramidal neurons. The fast component appeared in dendritic locations near the input sites in response to synaptic activation, and signal onset at the soma was delayed by a few milliseconds compared with that at the input sites. Local perfusion of a Na(+) channel blocker near the soma eliminated the fast component at the soma, but had no effect on the fast component at the input sites. Our results indicate that dendritic spikes can be initiated in dendrites near the input site and propagate orthodromically toward the proximal dendrites and the soma. PMID- 12732237 TI - Presynaptic and postsynaptic GABAA receptors in rat suprachiasmatic nucleus. AB - The mammalian suprachiasmatic nucleus (SCN), the brain's circadian clock, is composed mainly of GABAergic neurons, that are interconnected via synapses with GABA(A) receptors. Here we report on the subcellular localization of these receptors in the SCN, as revealed by an extensively characterized antibody to the alpha 3 subunit of GABA(A) receptors in conjunction with pre- and postembedding electron microscopic immunocytochemistry. GABA(A) receptor immunoreactivity was observed in neuronal perikarya, dendritic processes and axonal terminals. In perikarya and proximal dendrites, GABA(A) receptor immunoreactivity was expressed mainly in endoplasmic reticulum and Golgi complexes, while in the distal part of dendrites, immunoreaction product was associated with postsynaptic plasma membrane. Many GABAergic axonal terminals, as revealed by postembedding immunogold labeling, displayed GABA(A) receptor immunoreactivity, associated mainly with the extrasynaptic portion of their plasma membrane. The function of these receptors was studied in hypothalamic slices using whole-cell patch-clamp recording of the responses to minimal stimulation of an area dorsal to the SCN. Analysis of the evoked inhibitory postsynaptic currents showed that either bath or local application of 100 microM of GABA decreased GABAergic transmission, manifested as a two-fold increase in failure rate. This presynaptic effect, which was detected in the presence of the glutamate receptor blocker 6-cyano-7 nitroquinoxaline-2,3-dione and the selective GABA(B) receptor blocker CGP55845A, appears to be mediated via activation of GABA(A) receptors. Our results thus show that GABA(A) receptors are widely distributed in the SCN and may subserve both pre- and postsynaptic roles in controlling the mammalian circadian clock. PMID- 12732238 TI - Integrins regulate neuronal neurotrophin gene expression through effects on voltage-sensitive calcium channels. AB - Integrin adhesion receptors regulate gene expression during growth and differentiation in various cell types. Recent work, implicating integrins in functional synaptic plasticity, suggest they may have similar activities in adult brain. The present study tested if integrins binding the arginine-glycine aspartate (RGD) matrix sequence regulate neurotrophin and neurotrophin receptor gene expression in cultured hippocampal slices. The soluble RGD-containing peptide glycine-arginine-glycine-aspartate-serine-proline (GRGDSP) increased neurotrophin mRNA levels in transcript- and subfield-specific fashions. Integrin ligand effects were greatest for brain-derived neurotrophic factor transcripts I and II and barely detectable for transcript III. In accordance with increased nerve growth factor mRNA levels, GRGDSP increased c-fos expression as well. In contrast, growth-associated protein-43, amyloid precursor protein and fibroblast growth factor-1 mRNAs were not elevated. Ligand effects on brain-derived neurotrophic factor transcript II and c-fos mRNA did not depend on the integrity of the actin cytoskeleton, neuronal activity, or various signaling pathways but were blocked by L-type voltage-sensitive calcium-channel blockers. These results indicate that in mature hippocampal neurons integrin engagement regulates expression of a subset of growth-related genes at least in part through effects on calcium influx. Accordingly, these synaptic adhesion receptors may play the same role in maintaining an adult, differentiated state in brain as they do in other tissues and changes in integrin activation and/or engagement may contribute to dynamic changes in neurotrophin expression and to neuronal calcium signaling. PMID- 12732239 TI - Estradiol inhibits atp-induced intracellular calcium concentration increase in dorsal root ganglia neurons. AB - Estrogen has been implicated in modulation of pain processing. Although this modulation occurs within the CNS, estrogen may also act on primary afferent neurons whose cell bodies are located within the dorsal root ganglia (DRG). Primary cultures of rat DRG neurons were loaded with Fura-2 and tested for ATP induced changes in intracellular calcium concentration ([Ca(2+)](i)) by fluorescent ratio imaging. ATP, an algesic agent, induces [Ca(2+)](i) changes via activation of purinergic 2X (P2X) type receptors and voltage-gated Ca(2+) channels (VGCC). ATP (10 microM) caused increased [Ca(2+)](i) transients (226.6+/ 16.7 nM, n = 42) in 53% of small to medium DRG neurons. A 5-min incubation with 17 beta-estradiol (100 nM) inhibited ATP-induced [Ca(2+)](i) (164+/-14.6 nM, P<0.05) in 85% of the ATP-responsive DRG neurons, whereas the inactive isomer 17 alpha-estradiol had no effect. Both the mixed agonist/antagonist tamoxifen (1 microM) and specific estrogen receptor antagonist ICI 182780 (1 microM) blocked the estradiol inhibition of ATP-induced [Ca(2+)](i) transients. Estradiol coupled to bovine serum albumin, which does not diffuse through the plasma membrane, blocked ATP-induced [Ca(2+)](i), suggesting that estradiol acts at a membrane associated estrogen receptor. Attenuation of [Ca(2+)](i) transients was mediated by estrogen action on VGCC. Nifedipine (10 microM), an L-type VGCC antagonist mimicked the effect of estrogen and when co-administered did not increase the estradiol inhibition of ATP-induced [Ca(2+)](i) transients. N- and P-type VGCC antagonists omega-conotoxin GVIA (1 microM) and omega-agatoxin IVA (100 nM), attenuated the ATP-induced [Ca(2+)](i) transients. Co-administration of these blockers with estrogen induced a further decrease of the ATP-induced [Ca(2+)](i) flux. Together, these results suggest that although ATP stimulation of P2X receptors activates L-, N-, and P-type VGCC, estradiol primarily blocks L-type VGCC. The estradiol regulation of this ATP-induced [Ca(2+)](i) transients suggests a mechanism through which estradiol may modulate nociceptive signaling in the peripheral nervous system. PMID- 12732240 TI - Closed-head minimal traumatic brain injury produces long-term cognitive deficits in mice. AB - Victims of minimal traumatic brain injury (mTBI) do not show clear morphological brain defects, but frequently suffer lasting cognitive deficits, emotional difficulties and behavioral disturbances. In the present study we adopted a non invasive closed-head weight-drop mouse model to produce mTBI. We examined the effects of 20, 25, or 30 g weight drop 7, 30, 60 and 90 days following injury on mice's ability to perform the Morris water maze. The mice suffered profound long lasting learning and memory deficits that were force- and time-dependent. Although the injured mice could acquire the task, they could not improve their initial escape latency by more than 50%, while normal mice improved by up to 450% (P<0.001). In order to directly compare the learning ability of individual mice following our mTBI we have devised a new measure which we term learning rate. We define learning rate as the rate the mouse improved its own performance in consecutive trials in a given experimental day. The learning rate of control mice increased linearly throughout the testing period with a slope of approximately 0.9. Injured mice that sustained 20 and 25 g weight drop could also improve their learning rate linearly but with a slope of only 0.2. Mice who sustained 30 g weight drop could not improve their learning rate linearly and reached a plateau after the third experimental learning day. These results indicate that the severity of injury may correlate with the degree of integration of the learning task. These cognitive deficits occurred without any other clear neurological damage, no evident brain edema, no notable damage to the blood-brain barrier and no early anatomical changes to the brain (observed by magnetic resonance imaging imaging). These results demonstrate that persistent deficits of cognitive learning abilities in mice, similar to those observed in human post-concussive syndrome, can follow mTBI without any anatomical damage to the brain and its surrounding tissue. PMID- 12732241 TI - Differential expression and subcellular distribution of dystrophin Dp71 isoforms during differentiation process. AB - Dp71 is the major product of the Duchenne muscular dystrophy gene in the brain. In order to study the function of Dp71 in the nervous system we examined the expression of Dp71 isoforms in PC12 rat pheochromocytoma cell line, a well established system to study neuronal differentiation. We show by reverse transcriptase-polymerase chain reaction and Western blot assays that PC12 cells express two Dp71 isoforms. One isoform lacks exon 71 and the other isoform lacks exons 71 and 78 (Dp71d and Dp71f isoforms respectively). Nerve growth factor induced neuronal differentiation of PC12 cells results in differential regulation of the expression and subcellular localization of Dp71 isoforms: a) the amount of Dp71f protein increases nine-fold in total extracts while Dp71d increases up to seven-fold in nuclear extracts; b) Dp71f relocates from the cytoplasm to neuritic processes, being prominent at varicosities and the growth cone; c) Dp71d relocates almost entirely to the nucleus and is detected to a lower extent in the cytoplasm and neuritic processes. Dp71f co-localizes with beta-dystroglycan and synaptophysin while Dp71d co-localizes with beta-dystroglycan in the nucleus. Dp71d accumulates at cell-cell contacts where Dp71f is absent. These results suggest that Dp71d and Dp71f associate with different subcellular complexes and therefore may have distinct functions in PC12 cells. PMID- 12732242 TI - Neuronal activity regulates GABAA receptor subunit expression in organotypic hippocampal slice cultures. AB - The postnatal expression of GABA(A) receptor subunit mRNAs in the rat brain, including the hippocampus, exhibits a unique temporal and regional developmental profile in vivo, which may be altered by external stimuli. Using the in situ hybridization technique we have now studied the in vitro expression of alpha1,alpha2, alpha 4, alpha 5, beta 1, beta 3, gamma 2, and gamma 3 subunit mRNAs of GABA(A) receptors in organotypic hippocampal slices cultured for 7 days. To find out whether neuronal activity regulates the subunit expression, a subset of cultures was chronically treated either with a GABA(A) receptor antagonist picrotoxin, or by a non-N-methyl-D-aspartate (non-NMDA)-receptor antagonist 6,7 dinitroquinoxaline-2,3-dione (DNQX). In untreated control cultures, the expression pattern of the subunits varied regionally, the most abundantly expressed subunits being alpha 2 and alpha 5 in all subregions. All studied subunits were expressed in CA3a/b and CA1, whereas in CA3c and in granule cells of the dentate gyrus (DG) no signal of alpha 4 and gamma 3 was detected. The drug treatment differently affected the regional subunit expression. In picrotoxin treated cultures, the expression of alpha1, alpha 5 and gamma 2 mRNAs was significantly increased in pyramidal cell layers, and in DNQX-treated cultures the expression of alpha2 mRNA in CA3c and DG, and that of beta1 in DG. Changes in the expression of GABA(A) receptor subunit mRNAs in treated cultures suggest that neuronal activity can regulate their regional expression in vitro. Since the expression profile in untreated control cultures closely resembled that observed earlier in vivo, organotypic hippocampal slice cultures could serve as a good model system to study the regulatory mechanisms of receptor expression under well controlled experimental conditions in the developing hippocampus. PMID- 12732243 TI - Differential regulation of corticosteroid receptors by monoamine neurotransmitters and antidepressant drugs in primary hippocampal culture. AB - Hyperactivity of the hypothalamic-pituitary-adrenal axis is a characteristic feature of depressive illness. The centrally located corticosteroid receptors, the glucocorticoid and mineralocorticoid receptors, are thought to be important modulators of this axis and changes in the levels of these receptors, particularly in the hippocampus, may underlie the hyperactivity observed. Various antidepressant drugs increase hippocampal mineralocorticoid and glucocorticoid receptor levels in vivo. These effects are thought to be mediated via alterations in monoaminergic neurotransmission. We examined whether serotonin (5HT) and noradrenaline (NA) have direct effects on glucocorticoid receptor and mineralocorticoid receptor expression in primary hippocampal neurones, and whether antidepressants also exert direct effects on target neurones. Exposure of hippocampal cells to 5HT for 4 days increased both glucocorticoid and mineralocorticoid receptor mRNA and protein expression. The induction of mineralocorticoid receptor mRNA was completely blocked by the 5HT(7) receptor antagonist SB 269970. In contrast glucocorticoid receptor induction was insensitive to the 5HT(7) receptor, whilst studies with the 5HT(1A) receptor agonist 8-hydroxy-2-(di-n-proplamino) tetralin hydrochloride and the 5HT(1A) receptor antagonist N-[2-[4-2-[O-methoxyphenyl)-1-piperazinyl]ethyl]-N-(2 pyridinyl) cyclohexane carboxamide trihydrochloride (WAY 100635) suggest a partial role for 5HT(1A) receptors in hippocampal glucocorticoid receptor regulation. Treatment with NA for 4 days also increased glucocorticoid receptor expression but had no effect on mineralocorticoid receptor expression. This was blocked by propanolol suggesting action via beta-adrenergic receptors. Similarly to NA, fluoxetine and amitriptyline also selectively increased glucocorticoid receptor mRNA and protein levels over this time course. However, glucocorticoid receptor induction by fluoxetine or amitriptyline was not blocked by WAY 100635 or propanolol. These results show that 5HT, NA and antidepressants act directly but via distinct mechanisms on hippocampal neurones to regulate mineralocorticoid and glucocorticoid receptor expression. Thusly, manipulation of neurotransmitter or antidepressant levels in the brain may aid in reversing hypothalamic-pituitary adrenal axis hyperactivity by restoring hippocampal corticosteroid receptor balance. PMID- 12732244 TI - Role of alpha-synuclein in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced parkinsonism in mice. AB - In humans, mutations in the alpha-synuclein gene or exposure to the neurotoxin 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) produce Parkinson's disease with loss of dopaminergic neurons and depletion of nigrostriatal dopamine. alpha Synuclein is a vertebrate-specific component of presynaptic nerve terminals that may function in modulating synaptic transmission. To test whether MPTP toxicity involves alpha-synuclein, we generated alpha-synuclein-deficient mice by homologous recombination, and analyzed the effect of deleting alpha-synuclein on MPTP toxicity using these knockout mice. In addition, we examined commercially available mice that contain a spontaneous loss of the alpha-synuclein gene. As described previously, deletion of alpha-synuclein had no significant effects on brain structure or composition. In particular, the levels of synaptic proteins were not altered, and the concentrations of dopamine, dopamine metabolites, and dopaminergic proteins were unchanged. Upon acute MPTP challenge, alpha-synuclein knockout mice were partly protected from chronic depletion of nigrostriatal dopamine when compared with littermates of the same genetic background, whereas mice carrying the spontaneous deletion of the alpha-synuclein gene exhibited no protection. Furthermore, alpha-synuclein knockout mice but not the mice with the alpha-synuclein gene deletion were slightly more sensitive to methamphetamine than littermate control mice. These results demonstrate that alpha-synuclein is not obligatorily coupled to MPTP sensitivity, but can influence MPTP toxicity on some genetic backgrounds, and illustrate the need for extensive controls in studies aimed at describing the effects of mouse knockouts on MPTP sensitivity. PMID- 12732245 TI - Long-term regulation of N-methyl-D-aspartate receptor subunits and associated synaptic proteins following hippocampal synaptic plasticity. AB - Synaptic plasticity in the dentate gyrus is dependent on activation of the N methyl-D-aspartate (NMDA)-subtype of glutamate receptors. In this study, we show that synaptic plasticity in turn regulates NMDA receptors, since subunits of the NMDA receptor complex are bidirectionally and independently regulated in the dentate gyrus following activation of perforant synapses in awake animals. Low frequency stimulation that produced a mild synaptic depression resulted in a decrease in the NMDA receptor subunits NR1 and NR2B 48 h following stimulation. High-frequency stimulation that produced long-term potentiation resulted in an increase in NR1 and NR2B at the same time point. Further investigations revealed that in contrast to NR2B, NR1 levels increased gradually after long-term potentiation induction, reaching a peak level at 48 h, and were insensitive to the competitive NMDA receptor antagonist 3-3(2-carboxypiperazin-4-yl) propyl-1 phosphate. The increased levels of NR1 and NR2B at 48 h were found associated with synaptic membranes and with increased NMDA receptor-associated proteins, postsynaptic density protein 95, neuronal nitric oxide synthase and Ca(2+)/calmodulin-dependent protein kinase II, alpha subunit. These data suggest that the persistence of long-term potentiation is associated with an increase in the number of NMDA receptor complexes, which may be indicative of an increase in synaptic contact area. PMID- 12732246 TI - Antinociceptive effect of calcitonin gene-related peptide in the central nucleus of amygdala: activating opioid receptors through amygdala-periaqueductal gray pathway. AB - The central nucleus of amygdala (CeA) plays an important role in pain regulation. Calcitonin gene-related peptide (CGRP)-like immunoreactive fibers and CGRP receptors are distributed densely in CeA. The present study was performed to elucidate the role of CGRP in nociceptive regulation in the CeA of rats. Intra CeA injection of CGRP induced dose-dependent increases in the hind-paw withdrawal latency tested by hotplate test and Randall Selitto Test, indicating an antinociceptive effect of CGRP in CeA. Furthermore, the antinociceptive effect of CGRP was blocked by intra-CeA administration of the CGRP receptor antagonist CGRP8-37, suggesting that CGRP receptor1 is involved in the CGRP-induced antinociception. The CGRP-induced antinociception was attenuated by s.c. injection of the opioid antagonist naloxone, suggesting an involvement of endogenous opioid systems in CGRP-induced antinociception. Moreover, it was demonstrated that opioid receptors in the periaqueductal gray, but not in CeA, contributed to the CGRP-induced antinociception, indicating the importance of the pathway between CeA and the periaqueductal gray in CGRP-induced antinociception. Combining retrograde fluorescent tracing with immunohistochemistry, we found that met-enkephalinergic neurons were innervated by CGRP-containing terminals in CeA. Furthermore, most neurons in the CeA retrogradely traced from the periaqueductal gray were contacted by CGRP-containing terminals and some of them were surrounded by characteristic basket-like structures formed by the terminals, suggesting that CGRP innervates the neurons which project from CeA to the periaqueductal gray. The results indicate that CGRP activates the met-enkephalinergic neurons, which project from CeA to the periaqueductal gray, producing antinociceptive effect in rats. PMID- 12732247 TI - Continuous low-dose treatment with brain-derived neurotrophic factor or neurotrophin-3 protects striatal medium spiny neurons from mild neonatal hypoxia/ischemia: a stereological study. AB - This study aimed to investigate whether continuous, low-dose, intracerebral infusion of either brain-derived neurotrophic factor (BDNF) or neurotrophin-3 (NT 3) could protect against striatal neuronal loss in mild neonatal hypoxic/ischaemic brain injury. Continuous, low-dose, intracerebral treatment is likely to minimise unwanted side effects of a single high dose and lengthen the time window for neuroprotection. A milder, albeit brain damage-inducing, hypoxic/ischaemic injury paradigm was used since this situation is likely to produce the highest survival rates and thus the greatest prevalence. Anaesthetised postnatal day 7 rats were each stereotaxically implanted with a brain infusion kit connected to a micro-osmotic pump. The pump continuously infused either BDNF (4.5 microg/day), NT-3 (12 microg/day), or vehicle solution into the right striatum for 3 days from postnatal day 7. The intrastriatal presence of BDNF or NT-3 was verified immunohistochemically. On postnatal day 8, the rats underwent right common carotid artery ligation followed by hypoxic exposure for 1.5 h. Animals were weighed daily thereafter and killed 1 week later on postnatal day 14. The total number of medium spiny neurons within the right striatum was stereologically determined using an optical disector/Cavalieri combination. Other measures of neuroprotection such as brain weight and striatal infarct volume were also undertaken. BDNF or NT-3 significantly increased the total number of surviving medium spiny neurons by 43% and 33% respectively. This significant neuroprotection was not evident when brain weight, striatal volume, striatal infarct volume, and neuronal density measures for NT-3, were compared. These measures therefore missed the protective effect demonstrated by the total neuronal count. This suggests that stereological measurement of total neuronal number is needed to detect neuroprotection at 1 week after low-dose, continuously infused, neurotrophin treatment and mild hypoxic/ischaemic injury. The results also suggest that lower treatment doses may be more useful than previously thought. BDNF may be particularly useful since it fostered both neuroprotection and normal weight gain. The ability to rescue striatal neurons from death may contribute toward a potential short-term, low-dose neurotrophin treatment for mild perinatal hypoxic/ischaemic brain injury in humans. PMID- 12732248 TI - Distribution and binding parameters of GABAA receptors in the thalamic nuclei of Macaca mulatta and changes caused by lesioning in the globus pallidus and reticular thalamic nucleus. AB - Ascending output from the basal ganglia to the primate motor thalamus is carried by GABAergic nigro- and pallido-thalamic pathways, which interact with intrinsic thalamic GABAergic systems represented in primates by local circuit neurons and axons of the reticular thalamic nucleus. Disease-triggered pathological processes in the basal ganglia can compromise any of these pathways either directly or indirectly, yet the effects of basal ganglia lesioning on its thalamic afferent receiving territories has not been studied in primates. Two GABA(A) receptor ligands, [(3)H]muscimol and [(3)H]flunitrazepam, were used to study the distribution and binding properties of the receptor in intact monkeys, those with kainic acid lesions in the globus pallidus, and those with ibotenic acid lesions in the reticular nucleus using quantitative autoradiographic technique on cryostat sections of fresh frozen brain tissue. In control monkeys the binding affinities for [(3)H]muscimol averaged 50 nM in the thalamic nuclei and 86 nM in the basal ganglia while the binding densities varied (maximum density of binding sites [Bmax] range of 99.4-1000.1 fmol/mg of tissue). Binding affinities and Bmax values for [(3)H]flunitrazepam averaged 2.02 nM and 81-113 fmol/mg of tissue, respectively. Addition of 100-microM GABA increased average affinity to 1.35 nM whereas Bmax values increased anywhere from 1-50% in different nuclei. Zolpidem (100 nM) decreased binding by 68-80%. Bmax values for both ligands were decreased at the two survival times in both medial and lateral globus pallidus implying involvement of both nuclei in the lesion. Statistically significant, 40% decrease (P=0.055) of Bmax for [(3)H]muscimol was observed in the ventral anterior nucleus pars densicellularis (VAdc, the main pallidal projection territory in the thalamus) 1 week after globus pallidus lesioning and a 36% decrease (P=0.017) 4 months post-lesioning. In contrast, [(3)H]flunitrazepam Bmax values in the VAdc of the same animals were increased by 23% (P=0.021) at 1 week and 28% (P=0.005) 4 months postlesion, respectively. One week after the reticular nucleus lesioning, the binding densities of [(3)H]muscimol and [(3)H]flunitrazepam were decreased in the thalamic nuclei receiving projections from the lesioned reticular nucleus sector by approximately 50% (P<0.05) and 10-33% (P<0.05), respectively. The results suggest that different GABA(A) receptor subtypes are associated with different GABAergic systems in the thalamus which react differently to deafferentation. PMID- 12732249 TI - Facilitative role of prolactin-releasing peptide neurons in oxytocin cell activation after conditioned-fear stimuli. AB - Emotional stress activates oxytocin neurons in the hypothalamic supraoptic and paraventricular nuclei and stimulates oxytocin release from the posterior pituitary. Oxytocin neurons in the hypothalamus have synaptic contact with prolactin-releasing peptide (PrRP) neurons. Intracerebroventricular administration of PrRP stimulates oxytocin release from the pituitary. These observations raise the possibility that PrRP neurons play a role in oxytocin response to emotional stress. To test this hypothesis, we first examined expression of Fos protein, an immediate early gene product, in the PrRP neurons in the medulla oblongata after conditioned-fear stimuli. Conditioned-fear stimuli increased the number of PrRP cells expressing Fos protein especially in the dorsomedial medulla. In order to determine whether PrRP cells projecting to the supraoptic nucleus are activated after conditioned-fear stimuli, we injected retrograde tracers into the supraoptic nucleus. Conditioned-fear stimuli induced expression of Fos protein in retrogradely labeled PrRP cells in the dorsomedial medulla. Finally we investigated whether immunoneutralization of endogenous PrRP impairs oxytocin release after emotional stimuli. An i.c.v. injection of a mouse monoclonal anti-PrRP antibody impaired release of oxytocin but not of adrenocorticotrophic hormone or prolactin and did not significantly change freezing behavior in response to conditioned-fear stimuli. From these data, we conclude that PrRP neurons in the dorsomedial medulla that project to the hypothalamus play a facilitative role in oxytocin release after emotional stimuli in rats. PMID- 12732250 TI - Modulation of late phases of long-term potentiation in rat dentate gyrus by stimulation of the medial septum. AB - The prolonged maintenance of hippocampal long-term potentiation (LTP) seems to require heterosynaptic events during its induction. We have previously shown that stimulation of the basolateral nucleus of the amygdala (BLA) within a distinct time window can reinforce a transient early-LTP into a long-lasting late-LTP in the dentate gyrus (DG) in freely moving rats. We have shown that this reinforcement was dependent on beta-adrenergic and/or muscarinergic receptor activation and protein synthesis. However, since the BLA does not directly stimulate the DG the question remained by which inputs such heterosynaptic processes are triggered. We have now directly stimulated the medial septal pathway 15 min after induction of early-LTP in the DG and show that this input is capable of reinforcing early into late-LTP in a frequency-dependent manner. This septal reinforcement of DG LTP was dependent on beta-adrenergic receptor activation and protein synthesis. We suggest that the reinforcing effect of the BLA stimulation can, potentially, be mediated via the septal input to the DG, though it differs in its ability to induce or modulate functional plasticity. PMID- 12732251 TI - Regulation of neuropeptide mRNA expression in the basal ganglia by intrastriatal and intranigral transplants in the rat Parkinson model. AB - Previous studies have shown that intrastriatal transplants of dopamine (DA)-rich fetal ventral mesencephalic (VM) tissue can correct denervation-induced changes in the cellular expression of neuropeptide and receptor mRNAs in the rat Parkinson model. However, with the standard transplantation approach normalization of all cellular parameters has not been obtained. This may be due either to the incomplete striatal reinnervation achieved by these transplants, or to the ectopic placement of the grafts. In the present study we have used a microtransplantation approach to obtain a more complete reinnervation of the denervated striatum (20 micrograft deposits spread over the entire structure). Neurons were also implanted directly into the substantia nigra. In rats with multiple intrastriatal VM transplants the lesion-induced upregulation of mRNAs encoding for preproenkephalin (PPE), the D(2)-type DA-receptor, and the GABA synthesizing enzyme glutamic acid decarboxylase (GAD(67)) was normalized throughout the striatum, whereas the lesion-induced downregulation of preprotachykinin mRNA was unaffected. Intranigral grafts of either fetal DA-rich VM tissue or GABA-rich striatal tissue did not induce any changes in striatal neuropeptide and D(2)-receptor mRNA expression despite significant behavioral improvement. Comparison of the behavioral data with levels of neuropeptide expression showed that in rats with intrastriatal VM transplants a complete normalization of striatal PPE and GAD(67) mRNA expression did not translate into a complete recovery of spontaneous motor behaviors. The results show that extensive DA reinnervation of the host striatum by multiple VM microtransplants is insufficient to obtain full recovery of all lesion-induced changes at both the cellular and the behavioral level. A full reconstruction of the nigrostriatal pathway or, alternatively, modulation of basal ganglia function by grafting in non-striatal regions may be required to further improve the functional outcome in the DA-denervated brain. PMID- 12732252 TI - Creatine protects against the convulsive behavior and lactate production elicited by the intrastriatal injection of methylmalonate. AB - Methylmalonic acidemias are metabolic disorders caused by a severe deficiency of methylmalonyl-CoA mutase activity, which are characterized by neurological dysfunction, including convulsions. It has been reported that the accumulating metabolite, L-methylmalonic acid (MMA), inhibits succinate dehydrogenase leading to ATP depletion in vitro, and that the intrastriatal injection of MMA induces convulsions through secondary NMDA receptor stimulation. In this study we investigated the effect of creatine (1.2, 3.6 and 12.0 mg/kg, (i.p.), [DOSAGE ERROR CORRECTED] succinate (1.5 micromol/striatum) and MK-801 (3 nmol/striatum) on the convulsions and on the striatal lactate increase induced by MMA (4.5 micromol/striatum) in rats. The effect of creatine on the striatal phosphocreatine content and on MMA-induced phosphocreatine depletion was also evaluated. Creatine, succinate and MK-801 pretreatment decreased the number and duration of convulsive episodes and the lactate increase elicited by MMA. Creatine, but not succinate, prevented the convulsions and the lactate increase induced by the direct stimulation of NMDA receptors. Acute creatine administration increased the total striatal phosphocreatine content and prevented MMA-induced phosphocreatine depletion. Our results suggest that MMA increases lactate production through secondary NMDA receptor activation, and it is proposed that the anticonvulsant effect of creatine against MMA-induced convulsions may be due to an increase in the phosphocreatine content available for metabolic purposes. PMID- 12732253 TI - Inhibitory effects of D2 agonists by striatal injection on excessive release of dopamine and hyperactivity induced by Bay K 8644 in rats. AB - We investigated by means of behavioral and neurochemical studies the effects of either D(1) or D(2) agonist on excessive dopamine release and hyperactivity induced by the microinjection of Bay K 8644, and an L-type Ca(2+) channel stimulant, into the rat caudate putamen under a novel environmental condition. Hyperactivity (locomotor activity and rearing counts) and significant increases in extracellular dopamine levels induced by Bay K 8644 were concomitantly observed. D(1) agonist, SKF81297, administered into the caudate putamen did not block Bay K 8644-induced hyperactivity measured by monitoring both animal activity and increases in extracellular dopamine levels detected by microdialysis. Pretreatment with the D(2) agonists, bromocriptine, talipexole and pramipexole, into the caudate putamen significantly blocked Bay K 8644-induced hyperactivity for 45 min after Bay K 8644 administration, although the single administration of these agonists significantly potentiated locomotor activity and rearing behavior. Furthermore, these agonists significantly suppressed Bay K 8644 induced extracellular dopamine levels. Our results indicate that these D(2) agonists (1) act on postsynaptic neuronal D(2) receptors under conditions of normal or low dopamine release in the caudate putamen, and (2) act on presynaptic D(2) receptors (autoreceptors) when excessive levels of dopamine are released or hyperdopamine neuronal activity is induced. Consequently, the effect of D(2) agonists in the clinical treatment of Parkinson's disease may be due to stimulation of postsynaptic D(2) receptors rather than presynaptic autoreceptors. PMID- 12732254 TI - Topographic organization of projections from the amygdala to the visual cortex in the macaque monkey. AB - The topography of amygdaloid projections to the visual cortices in the macaque monkey was examined by injecting the fluorescent tracers Fast Blue and Diamidino Yellow at different locations in the occipital and temporal lobes and mapping the distribution of retrogradely labeled cells in the amygdala. Injections involving regions from rostral area TE to caudal area V1 all resulted in labeled cells within the basal nucleus of the amygdala. Relatively few double-labeled cells were observed even when the two injections were separated by less than 3 mm. The projections were rostrocaudally organized such that projections to caudal visual areas originated from dorsal and caudal portions of the magnocellular division of the basal nucleus while projections to more rostrally situated visual areas originated in more rostral and ventral portions of the basal nucleus. When injections involved rostral and medial portions of area TE, retrogradely labeled cells were observed in the accessory basal and lateral nuclei in addition to the basal nucleus. These data confirm that the amygdala gives rise to feedback projections to all levels of the "ventral stream" visual pathway. The projections do not appear to be diffusely distributed since few double-labeled cells were observed. The largest cells of the basal nucleus, those located in the magnocellular division, project the farthest in the visual system and innervate all occipital and temporal levels. The smaller cells, in the intermediate and parvicellular regions, project to more rostral and medial portions of the visual cortex. These results suggest that the amygdala may have substantial modulatory control over sensory processing at all stages of the ventral-stream visual cortical hierarchy. PMID- 12732255 TI - The GABAB receptor and recognition memory: possible modulation of its behavioral effects by the nitrergic system. AB - Functional activation of the GABA(B) receptor inhibits learning and memory processes, though discrepant findings, in this context, have also been reported. The present study was designed to investigate the role of the GABA(B) receptor on recognition memory in the rat. For this purpose, the effects induced by the GABA(B) agonist baclofen and the GABA(B) antagonist P-(3-aminopropyl)-P diethoxymethylphosphinic acid (CGP 35348) on memory were assessed by using the object-recognition task. In addition, the possible involvement of the nitrergic system on GABA(B) receptor's effects was also evaluated by using the same behavioral procedure. This is a working-memory paradigm based on the differential exploration of a new and familiar object. In a first dose-response study, baclofen (0.5, 2, and 4 mg/kg, i.p.), dose-dependently impaired animals' performance in this task, suggesting a modulation of acquisition and storage of information. CGP 35348 (100 and 300 mg/kg, i.p.), counteracted these baclofen induced performance deficits. The nitric oxide donor molsidomine, at the dose of 4 but not 2 mg/kg, i.p, successfully antagonized the deficits on cognition induced by the highest dose of baclofen (4 mg/kg). These results indicate a) that the GABA(B) receptor is involved in recognition memory and b) that an NO component modulates the effects of the GABA(B) receptor on learning and memory. PMID- 12732256 TI - Functional relation between interneuron input and population activity in the rat hippocampal cornu ammonis 1 area. AB - Inhibitory interneurons are important components of the cornu ammonis 1 (CA1) network, as they are strategically positioned to control network information transfer. We investigated in detail synaptic input to individual CA1 interneurons (mainly basket and bistratified cells) after the local circuit was activated through the Schaffer-Commissural pathway and related this input to the population activity of the pyramidal cells. Synaptic responses were measured under whole cell voltage clamp and population activity was determined from local field potentials. The synaptic input that was evoked in CA1 interneurons fell into two distinct groups. Disynaptic input with a long latency always started after the population spike with a mean latency of 3.0+/-0.3 ms (n=22) in respect to the peak of the population spike. This type of synaptic input to the interneurons was causally linked to the occurrence and amplitude of the population spike and most likely driven by CA1 pyramidal cells. Short-latency monosynaptic input occurred 0.8+/-0.2 ms (n=18) before the peak of the population spike. Its timing was strictly linked to the stimulus and showed significantly less jitter than long latency input. In the absence of a population spike only short-latency input could be observed. Whether an interneuron receives direct monosynaptic Schaffer input or disynaptic input from the pyramidal cell population determines when that interneuron will be recruited in the network after Schaffer collateral stimulation. In addition, we found that the relation between the strength of the synaptic input and the population activity was different for the two types of input. Short-latency monosynaptic input showed large sensitivity to input changes at stimulus intensities that evoked little activity in the pyramidal cell population. In contrast, the amplitude of the long-latency disynaptic input to the interneurons closely reflected the population activity and increased gradually with stimulus intensity. Interneurons receiving the first type of input may expand the input sensitivity of the network, while interneurons receiving the second type could be involved in overall normalization of the output of the CA1 network. Our results underscore the importance of knowledge of the input to an interneuron for the understanding of its inhibitory role in the network. PMID- 12732257 TI - Switching to cue-directed behavior: specific for ventral striatal dopamine but not ventral pallidum/substantia innominata gaba as revealed by a swimming-test procedure in rats. AB - In this study it was investigated whether ventral striatal dopamine-induced changes in switching to cue-directed behavioral patterns were funnelled via the rostral areas of the ventral pallidum/substantia innominata (VP/SI) complex and, if so, whether changes in switching to cue-directed behavioral patterns could be elicited in the VP/SI complex by manipulating GABAergic activity. To this end rats were bilaterally equipped with cannulae directed at the ventral striatum and/or rostral VP/SI complex and subjected to a swimming-test procedure for 6 min. Injections of the dopamine-releasing agent d-amphetamine (10 microg/0.5 microl per side) enhanced the number of different cue-directed behavioral patterns while they had no effect upon the number of different non-cue-directed behavioral patterns in line with previous studies (Life Sci - 1989 1697). This increase was attenuated by a low dose of the GABAa agonist muscimol (1 ng/0.5 microl) into the rostral VP/SI complex. This dose of muscimol when injected alone into the rostral VP/SI complex had no effect upon the number of different cue directed behavioral patterns. Only the lowest dose of the GABAa antagonist bicuculline (10-25 ng/0.5 microl per side) into the rostral VP/SI complex slightly, and in a non-d-amphetamine-like manner, increased the number of different cue-directed behavioral patterns while none of the doses had an effect on the number of different non-cue-directed behavioral patterns. Both injections of d-amphetamine into the ventral striatum and injections of bicuculline into the rostral VP/SI complex strongly increased motor activity in the 10-min period preceding the swimming test. We conclude from the data that switching to cue directed behavioral patterns is sensitive to manipulations with the dopaminergic activity in the ventral striatum but not with the GABAergic activity in the VP/SI complex although the VP/SI transmits it to other brain structures. In contrast motor activity is sensitive to manipulations with both ventral striatal dopamine and rostral VP/SI complex GABA. PMID- 12732258 TI - Regional brain cytochrome oxidase activity in beta-amyloid precursor protein transgenic mice with the Swedish mutation. AB - Cytochrome oxidase activity was examined in a transgenic mouse model of Alzheimer's disease with overexpression of the 751 amino acid isoform of beta amyloid precursor protein with the Swedish mutation under control of the murine thy-1 promoter. The neuritic plaques, abundantly localized in the hippocampus and anterior neocortical areas, showed a core devoid of enzymatic activity surrounded by higher cytochrome oxidase activity at the sites of the dystrophic neurites and activated glial cells. Quantitative measures, taken only in the healthy-appearing regional areas without neuritic plaques, were higher in numerous limbic and non limbic regions of transgenic mice in comparison with controls. Enzymatic activity was higher in the dentate gyrus and CA2-CA3 region of the hippocampus, the anterior cingulate and primary visual cortex, two olfactory structures, the ventral part of the neostriatum, the parafascicularis nucleus of the thalamus, and the subthalamic nucleus. Brainstem regions anatomically related with altered forebrain regions were more heavily labeled as well, including the substantia nigra, the periaqueductal gray, the superior colliculus, the medial raphe, the locus coeruleus and the adjacent parabrachial nucleus, as well as the pontine nuclei, red nucleus, and trigeminal motor nucleus. Functional brain organization is discussed in the context of Alzheimer's disease. Although hypometabolism is generally observed in this pathology, the increased cytochrome oxidase activity obtained in these transgenic mice can be the result of a functional compensation on the surviving neurons, or of an early mitochondrial alteration related to increased oxidative damage. PMID- 12732259 TI - Differential effects of intraspecific interactions on the striatal dopamine system in social and non-social voles. AB - We used in vivo microdialysis to examine the responses to intraspecific social interactions in the striatal dopamine systems of females of two vole species displaying vastly different social structures. Both highly social prairie voles and asocial meadow voles had similar increases in extracellular dopamine associated with mating. There was a species-specific effect of social condition on extracellular dihydroxyphenylacetic acid (DOPAC). Exposure to a conspecific male significantly decreased extracellular DOPAC in female prairie voles isolated for approximately 18 h during surgical recovery. Such decrease in DOPAC was not seen if females experienced continued isolation or if they were housed with a sibling during surgical recovery. No changes in extracellular DOPAC were seen in meadow voles after manipulations of social environment. Together, our data indicate that mating-associated dopamine release is independent from mating systems. However, species-specific patterns of extracellular DOPAC suggest that social isolation may be a more stressful stimulus for the social prairie vole than for the asocial meadow vole. PMID- 12732261 TI - Fluid shear stress and the vascular endothelium: for better and for worse. AB - As blood flows, the vascular wall is constantly subjected to physical forces, which regulate important physiological blood vessel responses, as well as being implicated in the development of arterial wall pathologies. Changes in blood flow, thus generating altered hemodynamic forces are responsible for acute vessel tone regulation, the development of blood vessel structure during embryogenesis and early growth, as well as chronic remodeling and generation of adult blood vessels. The complex interaction of biomechanical forces, and more specifically shear stress, derived by the flow of blood and the vascular endothelium raise many yet to be answered questions:How are mechanical forces transduced by endothelial cells into a biological response, and is there a "shear stress receptor"?Are "mechanical receptors" and the final signaling pathways they evoke similar to other stimulus-response transduction systems?How do vascular endothelial cells differ in their response to physiological or pathological shear stresses?Can shear stress receptors or shear stress responsive genes serve as novel targets for the design of diagnostic and therapeutic modalities for cardiovascular pathologies?The current review attempts to bring together recent findings on the in vivo and in vitro responses of the vascular endothelium to shear stress and to address some of the questions raised above. PMID- 12732260 TI - Inhibition of protein phosphatase 2A- and protein phosphatase 1-induced tau hyperphosphorylation and impairment of spatial memory retention in rats. AB - Tau hyperphosphorylation leads to formation of paired helical filament/neurofibrillary tangles, the hallmark lesion seen in Alzheimer's disease (AD) brain. An imbalanced regulation in protein kinases and protein phosphatases in the affected neurons is proposed to be a reasonable causative factor to the disease process. To verify the hypothesis, we have injected in the present study calyculin A, a potent and specific inhibitor of protein phosphatase (PP) 2A and PP1, into rat hippocampus bilaterally, thus reproduced an Alzheimer's-like deficiency in dephosphorylation system. It was found that calyculin A-injected rats developed lesions in spatial memory retention in Morris water maze test. At mean time, tau was hyperphosphorylated at Ser396/Ser404 (PHF-1) and Ser-262/Ser 356 (12E8) sites determined both by immunohistochemistry and Western blot. It is implicated that (1) PP2A and PP1 participate in the in vivo regulation of tau phosphorylation, and down-regulation of the two phosphatases will result in tau hyperphosphorylation; (2) hyperphosphorylation of tau at PHF-1 and 12E8 sites might be crucial to affect spatial memory in AD. PMID- 12732262 TI - Early assembly of cellular life. AB - Popular hypotheses that attempt to explain the origin of prebiotic molecules and cellular life capable of growth and division are not always agreed upon. In this manuscript, information on early bacterial life on Earth is examined using information from several disciplines. For example, knowledge can be integrated from physics, thermodynamics, planetary sciences, geology, biogeochemistry, lipid chemistry, primordial cell structures, cell and molecular biology, microbiology, metabolism and genetics. The origin of life also required a combination of elements, compounds and environmental physical-chemical conditions that allowed cells to assemble in less than a billion years. This may have been widespread in the subsurface of the early Earth located at microscopic physical domains. PMID- 12732264 TI - Cardiac mechano-electric feedback: past, present, and prospect. AB - Mechanical effects on heart rhythm have been known to the clinical community for well over a century, and documented cases include both arrhythmogenic and pro rhythmic consequences of mechanical stimulation. The intracardiac pathway that leads from changes in the cardiac mechanical environment to altered electrical activity is referred to as mechano-electric feedback (MEF). Fundamental research into the mechanisms underlying cardiac MEF is 'engineering-intensive', and much of the current insight would have been impossible without the introduction of novel techniques for the study of isolated cardiac cells. Clinical and basic research into MEF have developed over different time scales, often uninformed of each other, and utilizing disparate concepts and terminology. Bridging the gap between the two domains is not straightforward, as physicians and scientists tend to publish in different journals and attend different meetings. There is, however, a growing interest in 're-uniting' the clinic and basic MEF research, as witnessed by an increasing number of dedicated journal issues and international meetings, including events hosted by major European and American professional organisations such as the ESC and NASPE. Last year alone saw an international workshop on Cardiac MEF & Arrhythmias at Oxford, as well as dedicated sessions at NASPE's 23rd annual meeting in San Diego, CardioStim 2002 in Nice, and the UK Physiological Society meeting in Leeds. This volume of Progress in Biophysics and Molecular Biology incorporates clinical and basic science results, and it is fitting that its publication coincides with a special session on cardiac MEF at the 2003 meeting of NASPE. PMID- 12732263 TI - Allosteric activation of plasma membrane receptors--physiological implications and structural origins. AB - Allosteric modulation of receptors has physiological not just pharmacological significance. Thus, the chemical context in which an agonist signal is received can have a major impact on the nature of the physiological response by modifying receptor sensitivity and/or maximal activity-even the nature of the signalling response. In addition, recognising that an endogenous activator is the allosteric modulator of a known receptor, rather than the agonist of a novel receptor, has the potential to solve, in dramatic fashion, key physiological questions. What is an allosteric modulator and why are allosteric effects on receptors so diverse and frequently complex? What is the scope of allosteric effects? Can the existence of endogenous modulators be predicted from a receptor's amino acid sequence? How should screening for endogenous allosteric modulators be undertaken? These questions form the framework of this mini-review on physiological and structural aspects of receptor allostery. PMID- 12732265 TI - Evolutionary origins of mechanosensitive ion channels. AB - According to the recent revision, the universal phylogenetic tree is composed of three domains: Eukarya (eukaryotes), Bacteria (eubacteria) and Archaea (archaebacteria). Mechanosensitive (MS) ion channels have been documented in cells belonging to all three domains suggesting their very early appearance during evolution of life on Earth. The channels show great diversity in conductance, selectivity and voltage dependence, while sharing the property of being gated by mechanical stimuli exerted on cell membranes. In prokaryotes, MS channels were first documented in Bacteria followed by their discovery in Archaea. The finding of MS channels in archaeal cells helped to recognize and establish the evolutionary relationship between bacterial and archaeal MS channels and to show that this relationship extends to eukaryotic Fungi (Schizosaccharomyces pombe) and Plants (Arabidopsis thaliana). Similar to their bacterial and archaeal homologues, MS channels in eukaryotic cell-walled Fungi and Plants may serve in protecting the cellular plasma membrane from excessive dilation and rupture that may occur during osmotic stress. This review summarizes briefly some of the recent developments in the MS channel research field that may ultimately lead to elucidation of the biophysical and evolutionary principles underlying the mechanosensory transduction in living cells. PMID- 12732267 TI - Differential effects of stretch and compression on membrane currents and [Na+]c in ventricular myocytes. AB - Mechano-electrical feedback was studied in the single ventricular myocytes. A small fraction (approximately 10%) of the cell surface could be stretched or compressed by a glass stylus. Stretch depolarised, shortened the action potential and induced extra systoles. Stretch activated non-selective cation currents (I(ns)) showed a linear voltage dependence, a reversal potential of 0 mV, a pure cation selectivity, and were blocked by 8 microM Gd(3+) or 30 microM streptomycin. Stretch reduced Ca(2+) and K(+) (I(K)) currents. Local compression of broadwise attached cells activated I(K) but not I(ns). Cytochalasin D or colchicin, thought to disrupt the cytoskeleton, suppressed the mechanosensitivity of I(ns) and I(K). During stretch, the cytosolic sodium concentration increased with spatial heterogeneities, local hotspots with [Na(+)](c)>24 mM appeared close to surface membrane and t-tubules (pseudoratiometric imaging using Sodium Green fluorescence). Electronprobe microanalysis confirmed this result and indicated that stretch increased total sodium [Na] in cell compartments such as mitochondria, nuclear envelope and nucleus. Our results obtained by local stretch differ from those obtained by end-to-end stretch (literature). We speculate that channels may be activated not only by axial but also by shear stress, and, that stretch can activate channels outside the deformed sarcomeres via second messenger. PMID- 12732266 TI - Swelling-activated chloride channels in cardiac physiology and pathophysiology. AB - Characteristics and functions of the cardiac swelling-activated Cl current (I(Cl,swell)) are considered in physiologic and pathophysiologic settings. I(Cl,swell) is broadly distributed throughout the heart and is stimulated not only by osmotic and hydrostatic increases in cell volume, but also by agents that alter membrane tension and direct mechanical stretch. The current is outwardly rectifying, reverses between the plateau and resting potentials (E(m)), and is time-independent over the physiologic voltage range. Consequently, I(Cl,swell) shortens action potential duration, depolarizes E(m), and acts to decrease cell volume. Because it is activated by stimuli that also activate cation stretch activated channels, I(Cl,swell) should be considered as a potential effector of mechanoelectrical feedback. I(Cl,swell) is activated in ischemic and non-ischemic dilated cardiomyopathies and perhaps during ischemia and reperfusion. I(Cl,swell) plays a role in arrhythmogenesis, myocardial injury, preconditioning, and apoptosis of myocytes. As a result, I(Cl,swell) potentially is a novel therapeutic target. PMID- 12732268 TI - Stretch-modulation of second messengers: effects on cardiomyocyte ion transport. AB - In cardiomyocytes, mechanical stress induces a variety of hypertrophic responses including an increase in protein synthesis and a reprogramming of gene expression. Recently, the calcium signaling has been reported to play an important role in the development of cardiac hypertrophy. In this article, we report on the role of the calcium signaling in stretch-induced gene expression in cardiomyocytes. Stretching of cultured cardiomyocytes up-regulates the expression of brain natriuretic peptide (BNP). Intracellular calcium-elevating agents such as the calcium ionophore A23187, the calcium channel agonist BayK8644 and the sarcoplasmic reticulum calcium-ATPase inhibitor thapsigargin up-regulate BNP gene expression. Conversely, stretch-induced BNP gene expression is suppressed by EGTA, stretch-activated ion channel inhibitors, voltage-dependent calcium channel antagonists, and long-time exposure to thapsigargin. Furthermore, stretch increases the activity of calcium-dependent effectors such as calcineurin and calmodulin-dependent kinase II, and inhibitors of calcineurin and calmodulin dependent kinase II significantly attenuated stretch-induced hypertrophy and BNP expression. These results suggest that calcineurin and calmodulin-dependent kinase II are activated by calcium influx and subsequent calcium-induced calcium release, and play an important role in stretch-induced gene expression during the development of cardiac hypertrophy. PMID- 12732269 TI - Nitric-oxide-mediated regulation of cardiac contractility and stretch responses. AB - In the heart, nitric oxide (NO) is constitutively produced by the vascular and endocardial endothelium, the cardiomyocytes and the autonomic nerves. Whereas stimulation of NO release from the vascular endothelium has consistently been shown to quicken the onset of left ventricular (LV) relaxation and cause a small reduction in peak contraction, the role of myocardial NO production in regulating cardiac function appears to be more complex and controversial. Some studies have shown that non-isoform-specific inhibition of NO synthesis with L-arginine analogues has no effect on basal contraction in LV myocytes. However, others have demonstrated that stimulation of myocardial NO production can offset the increase in contraction in response to a rise in intracellular Ca(2+). Cardiac NO production is also activated by stretch and under these conditions NO has been shown to facilitate the Frank-Starling response and to contribute to the increase in intracellular Ca(2+) transients that mediates the slow increase in contraction in response to stretch (i.e., the Anrep effect). These findings suggest that NO can mediate diverse and even contrasting actions within the myocardium, a notion that is difficult to reconcile with the early description of NO as a highly reactive and diffusible molecule possessing minimal specificity in its interactions. The purpose of this short review is to revisit some of the 'controversial' aspects of NO-mediated regulation of myocardial function, taking into account our current understanding of how mammalian cells may target and regulate the synthesis of NO in such a way that NO can serve diverse physiological functions. PMID- 12732270 TI - Do stretch-induced changes in intracellular calcium modify the electrical activity of cardiac muscle? AB - Stretch of the myocardium influences the shape and amplitude of the intracellular Ca(2+)([Ca(2+)](i)) transient. Under isometric conditions stretch immediately increases myofilament Ca(2+) sensitivity, increasing force production and abbreviating the time course of the [Ca(2+)](i) transient (the rapid response). Conversely, muscle shortening can prolong the Ca(2+) transient by decreasing myofilament Ca(2+) sensitivity. During the cardiac cycle, increased ventricular dilation may increase myofilament Ca(2+) sensitivity during diastolic filling and the isovolumic phase of systole, but enhance the decrease in myofilament Ca(2+) sensitivity during the systolic shortening of the ejection phase. If stretch is maintained there is a gradual increase in the amplitude of the Ca(2+) transient and force production, which takes several minutes to develop fully (the slow response). The rapid and slow responses have been reported in whole hearts and single myocytes. Here we review stretch-induced changes in [Ca(2+)](i) and the underlying mechanisms. Myocardial stretch also modifies electrical activity and the opening of stretch-activated channels (SACs) is often used to explain this effect. However, the myocardium has many ionic currents that are regulated by [Ca(2+)](i) and in this review we discuss how stretch-induced changes in [Ca(2+)](i) can influence electrical activity via the modulation of these Ca(2+) dependent currents. Our recent work in single ventricular myocytes has shown that axial stretch prolongs the action potential. This effect is sensitive to either SAC blockade by streptomycin or the buffering of [Ca(2+)](i) with BAPTA, suggesting that both SACs and [Ca(2+)](i) are important for the full effects of axial stretch on electrical activity to develop. PMID- 12732271 TI - Stretch-induced excitation and action potential changes of single cardiac cells. AB - Mechanoelectric coupling (MEC) has been studied extensively in the heart at the tissue and organ levels, but to only a limited extent in single cells because of the technical challenges. New results are presented in which MEC was studied in 57 single frog ventricular myocytes that were held on both ends by glass holding pipettes. Axial stretch was applied either by displacement of the pipettes, or by a glass fiber around which the cell was wrapped, that was displaced in a pulsatile or sinusoidal fashion. Electrical activity of the cell was monitored either by active contraction, by intracellular action potentials, or by focal extracellular potentials. Of more than 350 stretches applied to 57 cells with amplitudes ranging from 3% to 35%, only 4 cases of mechanically induced stimulation were observed. In 252 stretches applied to 32 cells in which action potential duration (APD) was measured, no change >20% was observed, except in 3 cells in which APD increased by >100%, and in 2 cells with extended triggered activity. Thus, in contrast to studies in intact tissue, single frog ventricular myocytes are generally insensitive to direct axial stretch. However, robust mechanosensitive responses were observed in 7 of 57 ( approximately 12%) cells. The results of other single cell studies are reviewed, and the significance of differences in tissue-level and single cell results is discussed. PMID- 12732272 TI - Cardiac fibroblasts and the mechano-electric feedback mechanism in healthy and diseased hearts. AB - Cardiac arrhythmia is a serious clinical condition, which is frequently associated with abnormalities of mechanical loading and changes in wall tension of the heart. Recent novel findings suggest that fibroblasts may function as mechano-electric transducers in healthy and diseased hearts. Cardiac fibroblasts are electrically non-excitable cells that respond to spontaneous contractions of the myocardium with rhythmical changes of their resting membrane potential. This phenomenon is referred to as mechanically induced potential (MIP) and has been implicated in the mechano-electric feedback mechanism of the heart. Mechano electric feedback is thought to adjust the frequency of spontaneous myocardial contractions to changes in wall tension, which may result from variable filling pressure. Electrophysiological recordings of single atrial fibroblasts indicate that mechanical compression of the cells may activate a non-selective cation conductance leading to depolarisation of the membrane potential. Reduced amplitudes of MIPs due to pharmacological disruption of F-actin and tubulin suggest a role for the cytoskeleton in the mechano-electric signal transduction process. Enhanced sensitivity of the membrane potential of the fibroblasts to mechanical stretch after myocardial infarction correlates with depression of heart rates. It is assumed that altered electrical function of cardiac fibroblasts may contribute to the increased risk of post-infarct arrhythmia. PMID- 12732273 TI - Dynamic properties of stretch-activated K+ channels in adult rat atrial myocytes. AB - The effect of mechanical stress on the heart's electrical activity has been termed mechanoelectric feedback. The response to stretch depends upon the magnitude and the waveform of the stimulus, and upon the timing relative to the cardiac cycle. Stretch-activated ion channels (SACs) have been regarded as the most likely candidates for serving as the primary transducers of mechanical stress. We explored the steady state and dynamic responses of single channels in adult rat atrial cells using the patch clamp with a pressure clamp. Surprisingly, we only observed K(+)-selective SACs, probably of the 2P domain family. The channels were weakly outward rectifying with flickery bursts. In cell attached mode, the mean conductance was 74+/-14 and 65+/-16 pS for +60 and -60 mV, respectively (140 mM [K(+)](out), 2mM [Mg(2+)](out) and 0mM [Ca(2+)](out)). The latency of the response to pressure steps was 50-100 ms and the time to peak approximately 400 ms. About half of the channels in cell-attached patches showed adaptation/inactivation where channel activity declined to a plateau of 20-30% of peak in approximately 1s. The time dependent behavior of these SACs is generally consistent with whole-cell currents observed in chick and rat ventricular cells, although the net current was outward rather than inward. PMID- 12732274 TI - Mechano-electric feedback and atrial fibrillation. AB - Atrial fibrillation frequently occurs under conditions associated with atrial dilatation suggesting a role of mechano-electric feedback in atrial arrhythmogenesis. Although atrial arrhythmias may be due both to abnormal focal activity and reentrant mechanisms, the majority of sustained atrial arrhythmias have been ascribed to reentrant activity. Atrial stretch may contribute to focal arrhythmias by inducing afterdepolarizations and to reentrant arrhythmias by increasing the atrial surface, by shortening the refractory period and/or slowing the conduction velocity and by increasing their spatial dispersion. Experimental and clinical studies have demonstrated that changes in mechanical loading conditions may modulate the electrophysiological properties of the atria. These studies have, for the most part, involved the effects of acute stretch on atrial refractoriness. While studies in humans and intact animals yield divergent results due to the variety of loading conditions and neurohumoral influences, experimental studies in isolated preparations clearly show that atrial refractory period and action potential duration at early levels of repolarization shorten by acute atrial dilatation. Both experimental and human studies have shown that acute atrial stretch is arrhythmogenic and may induce triggered premature beats and atrial fibrillation. PMID- 12732275 TI - The role of atrial dilatation in the domestication of atrial fibrillation. AB - Numerous clinical investigations as well as recent experimental studies have demonstrated that atrial fibrillation (AF) is a progressive arrhythmia. With time paroxysmal AF becomes persistent and the success rate of cardioversion of persistent AF declines. Electrical remodeling (shortening of atrial refractoriness) develops within the first days of AF and contributes to the increase in stability of the arrhythmia. However, 'domestication of AF' must also depend on other mechanisms since the persistence of AF continues to increase after electrical remodeling has been completed. During the first days of AF in the goat, electrical and contractile remodeling (loss of atrial contractility) followed exactly the same time course suggesting that they are due to the same underlying mechanism. Contractile remodeling not only enhances the risk of atrial thrombus formation, it also enhances atrial dilatation by increasing the compliance of the fibrillating atrium. In goats with chronic AV-block atrial dilatation increased the duration of artificially induced AF-episodes but did not change atrial refractoriness or the AF cycle length. When AF was maintained a couple of days in these animals, a shortening of the atrial refractory period did occur. However, the AF cycle length did not decrease. Long lasting episodes of AF with a long AF cycle length and a wide excitable gap suggest that in this model AF is mainly promoted by conduction disturbances. Chronic atrial stretch induces activation of numerous signaling pathways leading to cellular hypertrophy, fibroblast proliferation and tissue fibrosis. The resulting electroanatomical substrate in dilated atria is characterized by increased non-uniform anisotropy and macroscopic slowing of conduction, promoting reentrant circuits in the atria. Prevention of electroanatomical remodeling by blockade of pathways activated by chronic atrial stretch therefore provides a promising strategy for future treatment of AF. PMID- 12732276 TI - Mechano-electrical feedback underlying arrhythmias: the atrial fibrillation case. AB - Mechanoelectrical feedback (MEF) has become firmly established as a mechanism in which mechanical forces experienced by myocardial tissue or cell membranes convey alterations in electrophysiologic characteristics of such tissue. Observations to date mainly concern mechanically induced changes in action potential duration, resting and active potential amplitude, enhanced pacemaker frequency, or afterdepolarizations. While some of these changes (i.e. after depolarizations) may give rise to premature beats, a role of MEF in explaining sustained ventricular tachyarrhythmias has so far been elusive. Here, we review recent findings showing that acute atrial dilatation facilitates atrial fibrillation (AF) and that two stretch-activated channel (SAC) blockers (gadolinium and GsMTx 4) are able to suppress stretch-facilitated AF. These findings strongly support a role of MEF and SACs in promoting sustained arrhythmias and point to a new class of antiarrhythmic drugs. PMID- 12732277 TI - Mechanically induced sudden death in chest wall impact (commotio cordis). AB - Sudden death due to nonpenetrating chest wall impact in the absence of injury to the ribs, sternum and heart is known as commotio cordis. Although once thought rare, an increasing number of these events have been reported. Indeed, a significant percentage of deaths on the athletic field are due to chest wall impact. Commotio cordis is most frequently observed in young individuals (age 4 18 years), but may also occur in adults. Sudden death is instantaneous or preceded by several seconds of lightheadedness after the chest wall blow. Victims are most often found in ventricular fibrillation, and successful resuscitation is more difficult than expected given the young age, excellent health of the victims, and the absence of structural heart disease. Autopsy examination is notable for the lack of any significant cardiac or thoracic abnormalities. In an experimental model of commotio cordis utilizing anesthetized juvenile swine, ventricular fibrillation can be produced by a 30 mph baseball strike if the strike occurred during the vulnerable period of repolarization, on the upslope of the T-wave. Energy of the impact object was also found to be a critical variable with 40 mph baseballs more likely to cause ventricular fibrillation than velocities less or greater than 40 mph. In addition, more rigid impact objects and blows directly over the center of the chest were more likely to cause ventricular fibrillation. Peak left ventricular pressure generated by the chest wall blow correlated with the risk of ventricular fibrillation. Activation of the K(+)(ATP) channel is a likely cause of the ventricular fibrillation produced by chest wall blows. Successful resuscitation is attainable with early defibrillation. PMID- 12732278 TI - Mechanical effects on arrhythmogenesis: from pipette to patient. AB - Mechanical stimuli delivered to the precordium can, if strong enough and timed at the beginning of the T-wave, induce ventricular premature beats or runs of ventricular tachycardia and even fibrillation. On the other hand, there are reports that a properly timed "chest thump" can terminate ventricular tachycardia, or can act as pacemaker stimuli during an episode of asystole. It is likely that in these cases mechanical energy is translated to an electrical stimulus. There are more subtle ways in which mechanical stimuli, mediated by stretch, can exert electrophysiological effects, and the most common name to describe these effects is mechanoelectrical feedback. Most studies have concentrated on acute stretch or dilatation, while the effects of chronic stretch, which may clinically be more important, are difficult to evaluate since they are accompanied by other factors, such as hypertrophy, heart failure, fibrosis, neurohumeral disturbances, and electrolyte abnormalities, all of which have arrhythmogenic effects. There are a number of ion channels that are activated following stretch. Stretch during diastole usually leads to a depolarization, resembling a delayed afterdepolarization, which may reach threshold and initiate a ventricular premature beat. Stretch during systole usually shortens the action potential, but action potential prolongation, resulting in early afterdepolarizations has been described as well. The arrhythmias during acute myocardial ischaemia occur in two phases: the 1A phase between 2 and 10 min following coronary artery occlusion, and the 1B phase between 18 and 30 min. Experiments will be described, indicating that the ventricular premature beats of the 1B phase, which may induce ventricular fibrillation, are caused by stretch of the border between ischaemic and normal myocardium. Briefly, 1B arrhythmias are much less frequent in the isolated perfused heart than in the heart in situ, but in working, ejecting isolated hearts, the number of 1B arrhythmias is similar to those in the in situ heart. The ventricular premature beats have a focal origin at the border, and they occur more often after a pause-induced potentiated contraction. PMID- 12732279 TI - Passive ventricular constraint. AB - Heart failure (HF) is a progressive degenerative and malignant syndrome with a large number of aetiologies including coronary artery disease, chronic hypertension, exposure to toxins, bacteria and viruses and in a significant percentage of HF patients, the causal mechanism is unclear. The HF trail of morbidity and mortality is well documented and is characterised by step-like periods of relative symptomatic stability, compensation, separated by decompensatory episodes. The homeostatic response to the decline in cardiac function is diverse and involves most organs. There is an increase in resting rate, intra-cardiac hormone production (catecholamines, aldosterone, etc.) and in particular structural changes occur with increased mass and dilatation (dilated cardiomyopathy, DCM). DCM is associated with decreased cardiac output, contractility and energy efficiency and an increase in pro-arrhythmia and conduction defects. Kass et al. (Circulation 91(9) (1995) 2314) first demonstrated in patients who had undergone a dynamic cardio-myoplasty procedure, that, preventing further dilatation in DCM was beneficial and that the improved cardiovascular status was largely independent of muscle stimulation. We hypothesised that this outcome could be achieved by implanting a fabric cardiac support device around both ventricles to the AV junction. Subsequently, it was shown by us and others (Kass et al., 1995) (Cardiovasc. Res. 44(3) (1999) 549); (Ann. Thorac. Surg. 70(4) (2000) 1275) (in different animal models of DCM) that passive ventricular constraint prevented further dilatation, initiated left ventricular volume reduction and reversed the decline in ejection fraction, mitral valve integrity and left ventricular contractility, when compared with untreated controls. Subsequent European and North American clinical trials in patients with DCM of varying aetiologies have shown equal promise and an absence of device related complications (Circulation 104(12 Suppl. 1) (2001) I270); (Ann. Thorac. Cardiovasc. Surg. 7(5) (2001) 278). The mechanisms behind this improvement have yet to be fully clarified however the support generated by the device upon the right and ventricular freewall would lower wall tension. Not only is passive ventricular constraint a very promising treatment modality for heart failure and DCM it should provide a useful research tool for the study of the role of ventricular dilatation in the progression of heart failure. PMID- 12732280 TI - Mechano-electric interactions in heterogeneous myocardium: development of fundamental experimental and theoretical models. AB - The heart is structurally and functionally a highly non-homogenous organ, yet its main function as a pump can only be achieved by the co-ordinated contraction of millions of ventricular cells. This apparent contradiction gives rise to the hypothesis that 'well-organised' inhomogeneity may be a pre-requisite for normal cardiac function. Here, we present a set of novel experimental and theoretical tools for the study of this concept. Heterogeneity, in its most condensed form, can be simulated using two individually controlled, mechanically interacting elements (duplex). We have developed and characterised three different types of duplexes: (i) biological duplex, consisting of two individually perfused biological samples (like thin papillary muscles or a trabeculae), (ii) virtual duplex, made-up of two interacting mathematical models of cardiac muscle, and (iii) hybrid duplex, containing a biological sample that interacts in real-time with a virtual muscle. In all three duplex types, in-series or in-parallel mechanical interaction of elements can be studied during externally isotonic, externally isometric, and auxotonic modes of contraction and relaxation. Duplex models, therefore, mimic (patho-)physiological mechano-electric interactions in heterogeneous myocardium at the multicellular level, and in an environment that allows one to control mechanical, electrical and pharmacological parameters. Results obtained using the duplex method show that: (i) contractile elements in heterogeneous myocardium are not 'independent' generators of tension/shortening, as their ino- and lusitropic characteristics change dynamically during mechanical interaction-potentially matching microscopic contractility to macroscopic demand, (ii) mechanical heterogeneity contributes differently to action potential duration (APD) changes, depending on whether mechanical coupling of elements is in-parallel or in-series, which may play a role in mechanical tuning of distant tissue regions, (iii) electro-mechanical activity of mechanically interacting contractile elements is affected by their activation sequence, which may optimise myocardial performance by smoothing intrinsic differences in APD. In conclusion, we present a novel set of tools for the experimental and theoretical investigation of cardiac mechano-electric interactions in healthy and/or diseased heterogeneous myocardium, which allows for the testing of previously inaccessible concepts. PMID- 12732281 TI - Effects of acute ventricular volume manipulation on in situ cardiomyocyte cell membrane configuration. AB - Effects of mechanical stimulation on cardiac electrical activity, gene expression, protein synthesis, and tissue remodelling have received increasing attention in recent years, as reviewed in this issue of PBMB. Little is known, though, about how changes in ventricular filling affect the cell configuration of cardiomyocytes in the ventricular wall. Here, we present first electron microscopic insight into changes in cardiomyocyte cell structure in situ during acute ventricular volume manipulation. Apart from confirming the anticipated ventricular volume-related changes in cardiomyocyte sarcomere length, there is evidence of (i) unfolding of 'slack' membrane, primarily from sarcolemmal invaginations near the Z-lines, and (ii) stretch-induced incorporation of sub membrane caveolae into the surface membrane. The functional relevance of these changes in cardiomyocyte membrane configuration-other than to cater for the length-dependent modulation of the cell surface to cell volume ratio-remains to be elucidated. PMID- 12732282 TI - Sarcomere length changes in a 3D mathematical model of the pig ventricles. AB - Measurements of the geometry and fibrous-sheet structure of the left and right ventricles of the pig heart are fitted with a finite element model. Mechanical changes during the heart cycle are computed by solving the equations of motion under specified ventricular boundary conditions and using experimentally defined constitutive laws for the active and passive material properties of myocardial tissue. The resulting patterns of deformation, such as axial torsion and changes in wall thickness and base-apex length, are consistent with experimental observations. The model can therefore be used to predict sarcomere length changes and other strain patterns throughout the myocardium and throughout the cardiac cycle. Here we present sarcomere length changes at a limited number of material points within the wall. Sarcomere length typically varies by 10% above and below the unloaded length; although under the boundary conditions imposed in the current model the midwall circumferentially oriented sarcomere lengths increased by up to 20% at end diastole. We provide web-access details for a downloadable software program designed to provide more extensive information on mechanical deformation, such as the principal strains and muscle fibre cross-sectional area changes during the cardiac cycle. PMID- 12732283 TI - Electrotonic cancellation of transmural electrical gradients in the left ventricle in man. AB - Myocardial cells isolated from different depths of the ventricular wall show substantial differences in action potential duration. Whether these electrophysiological differences are present in vivo when the cells are well coupled is a subject of ongoing controversy. This article provides a brief review and includes experimental evidence derived from patients undergoing cardiac surgery. PMID- 12732284 TI - Mechano-electric feedback and arrhythmias. AB - The mechanical state of the heart feeds back to modify cardiac rate and rhythm. Mechanical stretch of myocardial tissue causes immediate and chronic responses that lead to the common end point of arrhythmia. This review provides a brief summary of the author's personal choice of contributions that she considers have fostered our understanding of the role of mechano-electric feedback in arrhythmogenesis. Acute mechanical stretch reversibly depolarises the cell membrane and shortens the action potential duration. These electrophysiological changes are related to the activation of mechano-sensitive ion channels. Several different ion channels are involved in the sensing of stretch, among them K(+) selective, Cl(-)-selective, non-selective, and ATP-sensitive K(+) channels. Sodium and Ca(2+) entering the cells via non-selective ion channels are thought to contribute to the genesis of stretch-induced arrhythmia. Mechano-sensitive channels have been cloned from non-vertebrate and vertebrate species. Chronic stress on the heart activates gene expression in cardiomyocytes and non-myocytes. The signal transduction involves atrial natriuretic peptides and growth factors that initiate remodelling processes leading to hypertrophy which in turn may contribute to the electrical instability of the heart by increasing the responsiveness of mechano-sensitive channels. Selective block of these channels could provide some new form of treatment of mechanically induced arrhythmias, although at present there are no drugs available with sufficient selectivity. Detailed understanding of how mechanical strain on myocardial cells is translated into channel activation will allow to identify new targets for putative antiarrhythmic drugs. PMID- 12732285 TI - Oestrogen and progestin responses in human endometrium. AB - Our understanding of the mechanisms of the actions of oestrogens and progestins have evolved from the simple concept of nuclear receptor-mediated regulation of transcription to a highly sophisticated, finely tuned interplay between various coregulators, other signaling cascades and transcription factors. The net result of these complex regulatory mechanisms is a steroid-, cell-, or tissue-specific action of oestrogens and progestins, their antagonists or selective modulators of their receptors. In this review, we have attempted to shed some light on the regulation of the actions of oestrogens and progestins on the human endometrium. PMID- 12732286 TI - Glucocorticoid-induced maturation of glycoprotein galactosylation and fucosylation processes in the rat small intestine. AB - We determined the role of glucocorticoids in the maturation of glycoprotein galactosylation and fucosylation processes in the rat small intestine during postnatal development. Treatment of suckling rats with hydrocortisone (HC) increased activities of an O-glycan: galactosyltransferase, and of an alpha-1,2 fucosyltransferase, through transcriptional regulation of the FTB gene. The activities of a fucosyltransferase inhibitor and of the enzymes responsible for the synthesis and degradation of GDP-fucose were unaffected by the treatment, whereas a fall in the activity of alpha-L-fucosidase was observed. These changes were accompanied by the precocious appearance of alpha-1,2-fucose residues in complex glycan chains of brush-border membrane glycoproteins that normally appear after weaning, and with a trend to increase in alpha-1,2-fucose residues in mucins. Thus, treatment of suckling rats with hydrocortisone speeds up the maturation of glycoprotein galactosylation and fucosylation processes in the small intestine. The delayed increase in glucocorticoid levels induced by prolonged nursing, or the suppression of glucocorticoids by adrenalectomy (AD) before the normal rise in the hormone, both induced a delay in the increases in activities of the O-glycan: galactosyltransferase and alpha-1,2 fucosyltransferase observed normally after glucocorticoid enhancement. Thus, glucocorticoids might play at least a partial role in the maturation of glycoprotein glycosylation observed at weaning. PMID- 12732287 TI - Genistein inhibits vitamin D hydroxylases CYP24 and CYP27B1 expression in prostate cells. AB - In human prostate cancer cells, the availability of the steroid hormone 1,25 dihydroxyvitamin D(3) for antimitotic action is determined through the activity of the two enzymes CYP24 and CYP27B1, viz. 25-hydroxyvitamin D-24-hydroxylase and 25-hydroxyvitamin D-1alpha-hydroxylase. High performance liquid chromatography (HPLC) analysis of [(3)H]25(OH)D(3) metabolism in human prostate cancer DU-145 cells revealed that genistein and other isoflavonoids, such as dihydrogenistein and daidzein, as well as the antiestrogenic compound ICI 182,780, inhibited Vitamin D-metabolizing enzyme activities. Reverse transcriptase-polymerase chain reaction (RT-PCR) showed that only in case of genistein this was due to transcriptional inhibition of CYP24 and CYP27B1 gene expressions. In case of CYP27B1, reduction of gene activity involves histone deacetylation because genistein was inactive in the presence of the histone deactylase inhibitor trichostatin A. In contrast, under the same condition, CYP24 gene activity was largely suppressed. In summary, our results suggest that a combined effect of genistein and trichostatin A could increase the responsiveness of human prostate cancer cells to the antiproliferative action of 1,25-dihydroxyvitamin D(3). PMID- 12732288 TI - 4-ethoxymethylphenol: a novel phytoestrogen that acts as an agonist for human estrogen receptors. AB - Estrogen is the natural agonist of the estrogen receptor (ER). However, certain plant-derived compounds or phytoestrogens have been identified that mimic estrogens and act as agonists and/or antagonists of ERs, depending on subtype and target tissue. Using thin layer chromatography (TLC), gas chromatography-mass spectrometry (GC-MS), and proton nuclear magnetic resonance (1H-NMR), we identified a simple phenol, 4-ethoxymethylphenol (4EM), found in Maclura pomifera that acts as an agonist of ER-alpha and ER-beta in HeLa and MCF-7 cells. To study the effect of 4EM on ER-alpha and ER-beta activity, we performed transient transfection assays and showed that 4EM activated ER-dependent gene transcription in a dose-dependent manner on both ER subtypes and this activity was inhibited by trans-4-hydroxytamoxifen (4HT). Further, 4EM-mediated transcription in ER-alpha, like estrogen, was enhanced in the presence of coactivators, steroid receptor coactivator-1 (SRC-1), CREB binding proteins (CBP), and E6-associated protein (E6 AP). We found that 4EM was specific for ER and did not activate transcription of the progesterone receptor. PMID- 12732289 TI - Glucocorticoid regulation of human eosinophil gene expression. AB - Molecular analysis of steroid-regulated gene expression in freshly isolated human eosinophils is difficult due to the inherent high rate of spontaneous apoptosis and elevated levels of endogenous ribonucleases. To circumvent these limitations, we determined if the human eosinophilic cell line EoL-1 could serve as an in vitro model of glucocorticoid signaling. We found by optimizing growth conditions in low serum-containing media that dexamethasone (Dex) treatment of EoL-1 cells induced an apoptotic pathway that was inhibited by interleukin-5 (IL-5). Moreover, gene expression profiling using RNA from untreated EoL-1 cells and from freshly isolated human eosinophils identified 380 commonly expressed genes, including the eosinophil markers granule major basic protein, prostaglandin endoperoxide synthase 1 and arachidonate 15-lipoxygenase. Expression profiling was performed using EoL-1 cells that had been treated with dexamethasone for 0, 4, 12, 24 and 48h identifying 162 genes as differentially expressed. Two of the most highly upregulated genes based on expression profiling were the transcription factor Ets-2 and the MHC Class II genes (Q, R, and P). Expression of these genes in EoL-1 cells was shown to be dexamethasone-induced at the RNA and protein levels which is consistent with the known function of Ets-2 in controlling cell cycle progression and the role of MHC Class II antigens in mediating eosinophil functions. PMID- 12732290 TI - The effects of estradiol and selective estrogen receptor modulators on gene expression and messenger RNA stability in immortalized sheep endometrial stromal cells and human endometrial adenocarcinoma cells. AB - The purpose of this study was to identify an endometrial cell line that maintained the E2 up-regulation of estrogen receptor (ER) mRNA by enhanced message stability and to assess its dependence on ER protein. Estradiol (E2) effects on gene expression were measured in three cell lines: one immortalized from sheep endometrial stroma (ST) and two from human endometrial adenocarcinomas (Ishikawa and ECC-1). E2 up-regulated ER mRNA levels in ST and Ishikawa cells, but down-regulated ER mRNA levels in ECC-1 cells. E2 up-regulated progesterone receptor (PR), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and transforming growth factor-alpha (TGF-alpha) in both Ishikawa and ECC-1 cells. The selective estrogen receptor modulator ICI 182,780 antagonized the E2-induced up-regulation of ER and/or PR mRNA levels in all three cells, while another, GW 5638, antagonized the up-regulation of PR mRNA in Ishikawa and ECC-1 cells. In mechanistic studies, E2 had no effect on ER mRNA stability in ST cells and it destabilized ER mRNA in ECC-1 cells. Thus, Ishikawa cells appear to be the most physiologically relevant cell line in which to study the up-regulation of ER mRNA levels by enhanced mRNA stability. Its antagonism by ICI 182,780 reveals that ER protein is involved in this E2 response. PMID- 12732291 TI - A ginsenoside-Rh1, a component of ginseng saponin, activates estrogen receptor in human breast carcinoma MCF-7 cells. AB - We have examined the possibility that a component of Panax ginseng, ginsenoside Rh1, acts by binding to steroid hormone receptors such as receptors for estrogen, glucocorticoid, androgen, and retinoic acid. Ginsenoside-Rh1 activated the transcription of the estrogen-responsive luciferase reporter gene in MCF-7 breast cancer cells at a concentration of 50 microM. Activation was inhibited by the specific estrogen receptor antagonist ICI 182,780, indicating that the estrogenic effect of ginsenoside-Rh1 is estrogen receptor dependent. Ginsenoside-Rh1 induction of luciferase activity was dose-dependent in CV-1 cells transiently transfected with estrogen receptor and reporter plasmids. Next, we evaluated the ability of ginsenoside-Rh1 to induce the estrogen-responsive genes in MCF-7 cells. Ginsenoside-Rh1 increased c-fos and pS2 at the mRNA levels at 24h after treatment, although the effects were not as prominent as 17beta-estradiol. Western blot analysis showed that progesterone receptor protein was induced at 24h of treatment of ginsenoside-Rh1. However, ginsenoside-Rh1 failed to activate the glucocorticoid receptor, the androgen receptor, or the retinoic acid receptor in CV-1 cells transiently transfected with the corresponding steroid hormone receptors and hormone responsive reporter plasmids. These data support our hypothesis that ginsenoside-Rh1 acts as a weak phytoestrogen, presumably by binding and activating the estrogen receptor. PMID- 12732292 TI - Effect of antiestrogens and aromatase inhibitor on basal growth of the human breast cancer cell line MCF-7 in serum-free medium. AB - Antiestrogens are efficient inhibitors of estrogen-mediated growth of human breast cancer. Besides inhibiting estradiol-stimulated growth, antiestrogens may have a direct growth-inhibitory effect on estrogen receptor (ER) positive cells and thus be more efficient than aromatase inhibitors, which will only abrogate estrogen-dependent tumor growth. To address this issue, we have used the human breast cancer cell line MCF-7/S9 as a model system which is maintained in a chemically defined medium without serum and estrogen. The addition of estradiol results in an increase in cell growth rate. Thus, the MCF-7/S9 cell line is estrogen-responsive but not estrogen-dependent. Three different types of antiestrogens, namely tamoxifen, ICI 182,780 and EM-652 were found to exert a significant and dose-dependent inhibition of basal growth of MCF-7/S9 cells. The growth-inhibitory effect of the three antiestrogens was prevented by simultaneous estradiol treatment. Antiestrogen treatment also reduced the basal pS2 mRNA expression level, thus indicating spontaneous estrogenic activity in the cells. However, treatment with the aromatase inhibitor had no effect on basal cell growth, excluding that endogenous estrogen synthesis is involved in basal growth. These data demonstrate that in addition to their estrogen antagonistic effect, antiestrogens have a direct growth-inhibitory effect which is ER-mediated. Consequently, in the subset of ER positive breast cancer patients with estrogen independent tumor growth, antiestrogen therapy may be superior to treatment with aromatase inhibitors which only inhibit estrogen formation but do not affect cancer cell growth in the absence of estrogens. PMID- 12732293 TI - Norbormide enhances late steps of steroid-hormone synthesis in rat and mouse adrenal cortex. AB - Norbormide (N) is a vasoconstrictor agent, which acts selectively on the peripheral arteries of the rat, through the activation of the phospholipase C (PLC) cascade and the stimulation of Ca(2+) entrance in the vascular myocytes. Several endogenous vasoconstrictor agent (e.g. angiotensin-II (ANG-II) and endothelin-1 (ET-1)), that stimulate PLC pathway, are also able to enhance aldosterone secretion by the adrenal gland. Hence, we examined the effects of norbormide ((0.5, 1.0 or 5) x 10(-5)M) on corticosteroid-hormone secretion from adrenal slices of rats and mice. Quantitative HPLC assay showed that under basal conditions rat and mouse adrenal quarters secreted progesterone (PROG), 11 deoxycorticosterone (DOC), 18-hydroxy-DOC (18OH-DOC), corticosterone (CORT), 18 hydroxy-corticosterone (18OH-CORT) and aldosterone (ALDO), as well as large amounts of pregnenolone (PREG) when its metabolism was blocked by 10(-5)M cyanoketone. Norbormide concentration-dependently raised the secretion of all post-DOC steroids assayed, decreased progesterone and DOC production, and did not affect pregnenolone release. In conclusion, norbormide is able to enhance late steps of steroid synthesis, i.e. those leading to the transformation of DOC to corticosterone and aldosterone, without affecting early steps. This is an interesting finding because the other main endogenous adrenal secretagogues are known to stimulate both early and late steps of steroid synthesis. The mechanism underlying the selective activating action of norbormide on 11beta- and 18 hydroxylation remains to be investigated. PMID- 12732294 TI - Aromatase in skeletal muscle. AB - Aromatase gene expression and activity have been studied in human skeletal muscle. Using two separate rounds of RT-nested PCR, transcripts from the coding region of aromatase mRNA were detected in 32 of 34 samples. In terms of the non coding exon I, PCR product for I.4 was detected in 13 cases (38%), I.3 in 10 cases (29%), P.II in 6 cases (18%) and I.1 was not detected in any case. No transcripts for any studied variants of exon I were detected in 18 samples (53%). Aromatase activity was assessed using two different methodologies: in 19 cases by definitive product isolation (DPI) and in 42 cases by tritium-release assay (TRA). Using both methods detectable activity was present in 52% of cases. Average values for the cases with detectable activity were 2.2 fmol/mg protein/h for DPI and 6.5 fmol/mg protein/h for TRA. In the cohort studied by TRA, a positive correlation of aromatase activity with age of the donor was observed (r=0.34, P=0.03). In six cases paired comparison of aromatase activity in muscle and associated fat tissue were performed by DPI. When expressed per milligram of protein the activity was always higher in fat. However, this difference disappeared when activity was based on the gram of wet tissue. Taking into account bulk in the body it is concluded that muscle can be an important source of estrogens in men and post-menopausal women and its contribution to the circulating pool of estrogens may be comparable to that of adipose tissue. The nature of mRNA transcripts in muscle suggests that estrogen formation may be controlled by glucocorticoid- as well as cAMP-dependent promoters of the aromatase gene. PMID- 12732296 TI - RNA polymerase holoenzyme: structure, function and biological implications. AB - The past three years have marked the breakthrough in our understanding of the structural and functional organization of RNA polymerase. The latest major advance was the high-resolution structures of bacterial RNA polymerase holoenzyme and the holoenzyme in complex with promoter DNA. Together with an array of genetic, biochemical and biophysical data accumulated to date, the structures provide a comprehensive view of dynamic interactions between the major components of transcription machinery during the early stages of the transcription cycle. They include the binding of sigma factor to the core enzyme, and the recognition of promoter sequences and DNA melting by holoenzyme, transcription initiation and promoter clearance. PMID- 12732297 TI - Polarisation of prokaryotic chromosomes. AB - In many prokaryotes, asymmetrical mutational or selective pressures have caused compositional skews between complementary strands of replication arms, especially sensitive in the distribution of guanine and cytosine. In Escherichia coli, most of the guanine/cytosine skew is caused by mutation rates differing on leading and lagging strands, but contribution of skewed functionally important guanine-rich motifs (Chi and Rag sites), which control chromosome repair or positioning, is noticeable. Interference between replication and gene expression plays a minor role. The situation may be different in other bacteria. Studies of chromosome processing and bacterial taxonomy might profit from consideration of chromosome polarisation. PMID- 12732298 TI - Eyeing bacterial genomes. AB - The density of information in a bacterial genome allows its history, organization and encoded functions to be distilled into a single graphical representation. These features have made it possible to discern the forces acting in and on bacterial genomes at levels not attainable in eukaryotes. PMID- 12732299 TI - Uses and pitfalls of microarrays for studying transcriptional regulation. AB - Microarrays provide a powerful new tool for understanding the regulation of gene expression in bacteria. Many recent publications have used microarrays for identifying regulon members and stimulons that describe the complex organismal responses to environmental perturbations. The use of bioinformatics to identify DNA binding sites of transcription factors greatly facilitates the interpretation of these experiments. Understanding the transcriptome of an organism includes identifying all transcripts and mapping their 5' and 3' ends. High-density oligonucleotide arrays have enabled the identification of many new transcripts, including small RNAs and antisense RNAs. PMID- 12732301 TI - Motifs, modules and games in bacteria. AB - Global explorations of regulatory network dynamics, organization and evolution have become tractable thanks to high-throughput sequencing and molecular measurement of bacterial physiology. From these, a nascent conceptual framework is developing, that views the principles of regulation in term of motifs, modules and games. Motifs are small, repeated, and conserved biological units ranging from molecular domains to small reaction networks. They are arranged into functional modules, genetically dissectible cellular functions such as the cell cycle, or different stress responses. The dynamical functioning of modules defines the organism's strategy to survive in a game, pitting cell against cell, and cell against environment. Placing pathway structure and dynamics into an evolutionary context begins to allow discrimination between those physical and molecular features that particularize a species to its surroundings, and those that provide core physiological function. This approach promises to generate a higher level understanding of cellular design, pathway evolution and cellular bioengineering. PMID- 12732300 TI - Regulatory roles for small RNAs in bacteria. AB - Small RNAs can act to regulate both the synthesis of proteins, by affecting mRNA transcription, translation and stability, and the activity of specific proteins by binding to them. As a result of recent genome-wide screens, around 50 small RNAs have now been identified in Escherichia coli. These include many that require the RNA-binding protein Hfq for their activity; most of these RNAs act by pairing with their target mRNAs. Small RNAs can both positively and negatively regulate translation, can simultaneously regulate multiple mRNA targets, and can change the pattern of polarity within an operon. PMID- 12732302 TI - Function of the universally conserved bacterial GTPases. AB - The GTPase superfamily of cellular regulators is well represented in bacteria. A small number are universally conserved over the entire range of bacterial species. Such a pervasive taxonomic distribution suggests that these enzymes play important roles in bacterial cellular systems. Recent advances have demonstrated that bacterial GTPases are important regulators of ribosome function, and important for the distribution of DNA to daughter cells following cell division. In addition, the atomic structure of a unique GTPase, EngA, has recently been established. Unlike any other GTPase, EngA contains tandem GTP-binding domains. This structural study suggests that the GTPase cycles of the domains are regulated differentially in a manner that remains to be elucidated. PMID- 12732303 TI - The bacterial universal stress protein: function and regulation. AB - The universal stress protein A (UspA) superfamily encompasses an ancient and conserved group of proteins that are found in bacteria, Archea, fungi, flies and plants. The Escherichia coli UspA is produced in response to a large number of different environmental onslaughts and UspA is one of the most abundant proteins in growth-arrested cells. Although insights into the regulation of the E. coli uspA gene have been gained, the exact roles of the Usp proteins and Usp domains remain enigmatic; they appear, in some cases, to be linked to resistance to DNA damaging agents and to respiratory uncouplers. PMID- 12732304 TI - Coupling the initiation of chromosome replication to cell size in Escherichia coli. AB - Bacterial cells change size dramatically with change in growth rate, but the ratio between cell volume and the number of copies of the origin of chromosome replication (oriC) is roughly constant at the time of initiation of DNA replication at almost all growth rates. Recent research on the inactivation of initiator protein (DnaA) and depletion of DnaA pools by the high-affinity DnaA binding locus datA allows us to propose a simple model to explain the long standing question of how Escherichia coli couples DNA replication to cell size. PMID- 12732305 TI - Control of rRNA expression in Escherichia coli. AB - The control of ribosome synthesis has been a major focus in molecular biology for over 50 years. As protein synthesis is an essential, yet energetically costly, process, all cells (from bacteria to mammals) devote complex regulatory networks to fine-tune the expression of ribosomal RNA (and therefore ribosome synthesis) to the nutritional environment. In Escherichia coli, ribosomal RNA promoters are among the strongest in the cell and are regulated by trans-acting proteins (Fis and H-NS) and small molecules (guanosine 5'-diphosphate 3'-diphosphate and initiating nucleoside triphosphates). Recent work has dissected many of the molecular mechanisms responsible for the strength and regulation of rRNA promoters. PMID- 12732306 TI - Ribosome-tethered molecular chaperones: the first line of defense against protein misfolding? AB - Folding of many cellular proteins is facilitated by molecular chaperones. Analysis of both prokaryotic and lower eukaryotic model systems has revealed the presence of ribosome-associated molecular chaperones, thought to be the first line of defense against protein aggregation as translating polypeptides emerge from the ribosome. However, structurally unrelated chaperones have evolved to carry out these functions in different microbes. In the yeast Saccharomyces cerevisiae, an unusual complex of Hsp70 and J-type chaperones associates with ribosome-bound nascent chains, whereas in Escherichia coli the ribosome associated peptidyl-prolyl-cis-trans isomerase, trigger factor, plays a predominant role. PMID- 12732307 TI - Regulation by proteolysis in bacterial cells. AB - Regulation by proteolysis plays a major role in bacterial stress responses, the cell cycle and development. Key regulators of these processes are subject to conditional proteolysis that depends on complex cellular information processing. This information includes temporal and spatial cues, and recent research has revealed a striking potential for multiple signal integration. PMID- 12732308 TI - Regulation of the FecI-type ECF sigma factor by transmembrane signalling. AB - Induction of the ferric citrate transport genes of Escherichia coli K-12 involves a signalling cascade that starts at the cell surface and proceeds to the cytoplasm. Three specific proteins are involved: FecA in the outer membrane, FecR in the cytoplasmic membrane, and FecI in the cytoplasm. The binding of dinuclear ferric citrate to FecA causes substantial structural changes in FecA, triggering the signal cascade. The amino-proximal end of FecA interacts with the carboxy proximal end of FecR in the periplasm. FecR then transmits the signal across the cytoplasmic membrane into the cytoplasm and activates the FecI sigma factor, which binds to the RNA polymerase core enzyme and directs the RNA polymerase to the promoter upstream of the fecABCDE transport genes to initiate transcription. Transcription of the fecIR regulatory genes and the fec transport genes is repressed by the Fur protein loaded with Fe(2+). Therefore, transcription of the fec transport genes is subjected to double control: cells first detect iron deficiency and respond by synthesis of the regulatory proteins FecI and FecR, which initiate transcription of the fec transport genes, provided ferric citrate is available. FecI belongs to the extracytoplasmic function sigma factors, which are widespread among bacteria. With the recent sequencing of complete microbial genomes, it has become apparent that the FecIRA cascade is now a paradigm for the regulatory control of FecI family sigmas in Gram-negative bacteria. PMID- 12732309 TI - The role of Fe-S proteins in sensing and regulation in bacteria. AB - Fe-S clusters are key to the sensing and transcription functions of three transcription factors, FNR, IscR and SoxR. All three proteins were discovered in Escherichia coli but experimental data and bioinformatic predictions suggest that homologs of these proteins exist in other bacterial species, highlighting the widespread nature of Fe-S-dependent regulatory networks. In addition, the nearly ubiquitous citric acid cycle enzyme, aconitase, plays a role in translational regulation in E. coli and Bacillus subtilis when it loses its Fe-S cluster. Although these regulatory proteins have the common feature of containing an Fe-S cluster, they differ in the physiological signals that they respond to. Therefore, these regulatory factors provide insights into the chemical versatility of Fe-S clusters. PMID- 12732310 TI - From motility to virulence: Sensing and responding to environmental signals in Vibrio cholerae. AB - Sensing its changing environment is key for Vibrio cholerae when making the transition from an aquatic lifestyle to one more suited to a human host. An inverse correlation between motility and virulence gene expression has been reported, with the NADH : ubiquinone oxidoreductase system which powers motility by generating a sodium-motive force, playing a pivotal role. Recent studies have demonstrated that bile inhibits activity of the transcription factor ToxT, a protein responsible for direct activation of numerous virulence gene promoters. In addition, recent technological advances have allowed for the analysis of in vivo-induced genes and assessment of their timing of expression. Use of recombinase-based in vivo expression technology has revealed that the toxin coregulated pilus (a colonization factor) is expressed before cholera toxin. Components of an acid-tolerance response system have also been found using this method as well as signature-tagged mutagenesis. Finally, a role for quorum sensing in regulation of virulence gene expression has recently been established. PMID- 12732312 TI - The top genes: on the distance from transcript to function in yeast glycolysis. AB - With its abundant components and extensive study, the glycolytic pathway in the yeast Saccharomyces cerevisiae would appear ideal to obtain and reconcile the 'omes of transcript, protein, metabolite and flux. But to do so is challenging and, as is often the case, close correlation of gene expression and function is elusive, even in this organism. PMID- 12732311 TI - LuxS quorum sensing: more than just a numbers game. AB - Quorum sensing is a process of bacterial cell-to-cell communication involving the production and detection of extracellular signaling molecules called autoinducers. Quorum sensing allows populations of bacteria to collectively control gene expression, and thus synchronize group behavior. Processes controlled by quorum sensing are typically ones that are unproductive unless many bacteria act together. Most autoinducers enable intraspecies communication; however, a recently discovered autoinducer AI-2 has been proposed to serve as a 'universal signal' for interspecies communication. Studies suggest that AI-2 encodes information in addition to specifics about cell number. PMID- 12732314 TI - From measurements of metabolites to metabolomics: an 'on the fly' perspective illustrated by recent studies of carbon-nitrogen interactions. AB - Techniques for surveying metabolite levels provide a powerful tool for basic plant research and biotechnology. They offer the possibility of specialised measurements of specific classes of metabolites, profiling of a very broad range of low molecular weight compounds, and resolving the spatial and/or temporal localisation of selected key metabolites. Recent applications in the field of carbon-nitrogen interactions provide a framework to discuss how metabolite datasets are being utilised in the post-genomic era to characterise system responses, link transcript data to phenotypic responses, analyse underlying regulation mechanisms, and implement an in-context analysis of gene function. A major future challenge concerns the integration of the information gained from metabolite profiling into an accessible body of knowledge. PMID- 12732315 TI - Plant cell factories and mucosal vaccines. AB - Many advances continue to be made in the field of plant-derived vaccines. Plants have been shown capable of expressing a multicomponent vaccine that when orally delivered induces a T-helper cell subset 1 response and enables passive immunization. Furthermore, a plant-derived vaccine has been shown to protect against challenge in the target host. Increased antigen expression levels (up to 4.1% total soluble protein) have been obtained through transformation of the chloroplast genome. In view of these findings, plant-derived vaccines have been proved as valuable commodities to the world's health system; however, before their application, studies need to focus on optimization of immunization strategies and to investigate antigen stability. PMID- 12732316 TI - Plant cytochromes P450: tools for pharmacology, plant protection and phytoremediation. AB - Cytochromes P450 catalyse extremely diverse and often complex regiospecific and/or stereospecific reactions in the biosynthesis or catabolism of plant bioactive molecules. Engineered P450 expression is needed for low-cost production of antineoplastic drugs such as taxol or indole alkaloids and offers the possibility to increase the content of nutraceuticals such as phytoestrogens and antioxidants in plants. Natural products may serve important functions in plant defence and metabolic engineering of P450s is a prime target to improve plant defence against insects and pathogens. Herbicides, pollutants and other xenobiotics are metabolised by some plant P450 enzymes. These P450s are tools to modify herbicide tolerance, as selectable markers and for bioremediation. PMID- 12732317 TI - New genes in alkaloid metabolism and transport. AB - The biosynthetic pathway of plant alkaloids is composed of several distinct enzymes of varying substrate specificities. Homology-based cloning of candidate genes and their subsequent functional testing in heterologous expression systems are accelerating the pace at which the gene catalogues of alkaloid biosynthesis are expanding. Availability of diverse genes involved in the biosynthesis, catabolism, transport, and regulation of pharmaceutically important alkaloids should steadily advance our molecular understanding of alkaloid biology and will enable us to devise more rational strategies for metabolic engineering. PMID- 12732318 TI - Attracting friends to feast on foes: engineering terpene emission to make crop plants more attractive to herbivore enemies. AB - When attacked by herbivorous insects or mites, some plant species call on other arthropods for help. They emit mixtures of volatile compounds, dominated by terpenes, to attract carnivorous arthropods that prey on or parasitise herbivores and so reduce further damage. This fascinating defence strategy offers a new, environmentally friendly approach to crop protection. Using recent advances in the biochemistry and molecular genetics of terpene biosynthesis, it should now be possible to engineer crop plants that release terpenes for attracting herbivore enemies. By introducing or selectively altering the existing rate of terpene emission and composition, plant breeders could enable attacked plants to attract enemies and reduce additional herbivory, without compromising the effectiveness of other modes of defence. PMID- 12732319 TI - Deciphering plant-pathogen communication: fresh perspectives for molecular resistance breeding. AB - Activation of local and systemic plant defences in response to pathogen attack involves dramatic cellular reprogramming. Over the past 10 years many novel genes, proteins and molecules have been discovered as a result of investigating plant-pathogen interactions. Most attempts to harness this knowledge to engineer improved disease resistance in crops have failed. Although gene efficacy in transgenic plants has often been good, commercial exploitation has not been possible because of the detrimental effects on plant growth, development and crop yield. Biotechnology approaches have now shifted emphasis towards marker-assisted breeding and the construction of vectors containing highly regulated transgenes that confer resistance in several distinct ways. PMID- 12732320 TI - Molecular responses to drought, salinity and frost: common and different paths for plant protection. AB - Drought, high salinity and low temperature are major environmental factors that limit plant productivity. Plants respond and adapt to these stresses in order to survive. Signaling pathways are induced in response to environmental stress and recent molecular and genetic studies have revealed that these pathways involve many components. In this review, we highlight recent findings on the gene expression associated with stress responses and the signaling pathways that are either common or specific to the response. PMID- 12732321 TI - Genomics insights into symbiotic nitrogen fixation. AB - Following an interaction with rhizobial soil bacteria, legume plants are able to form a novel organ, termed the root nodule. This organ houses the rhizobial microsymbionts, which perform the biological nitrogen fixation process resulting in the incorporation of ammonia into plant organic molecules. Recent advances in genomics have opened exciting new perspectives in this field by providing the complete gene inventory of two rhizobial microsymbionts. The complete genome sequences of Mesorhizobium loti, the symbiont of several Lotus species, and Sinorhizobium meliloti, the symbiont of alfalfa, were determined and annotated in detail. For legume macrosymbionts, expressed sequence tag projects and expression analyses using DNA arrays in conjunction with proteomics approaches have identified numerous genes involved in root nodule formation and nitrogen fixation. The isolation of legume genes by tagging or positional cloning recently allowed the identification of genes that control the very early steps of root nodule organogenesis. PMID- 12732322 TI - Out of the woods: forest biotechnology enters the genomic era. AB - Trees represent a unique life form of upmost importance for mankind, as these organisms have developed a perennial lifestyle that produces the majority of terrestrial biomass. The difference between trees and annual plants is one of the main arguments behind the effort to sequence the entire genome of the poplar tree. This initiative is being backed up with a full-scale functional genomics effort on trees that will set a completely new agenda for forest research. PMID- 12732323 TI - From plant genomics to breeding practice. AB - New alleles are constantly accumulated during intentional crop selection. The molecular understanding of these alleles has stimulated new genomic approaches to mapping quantitative trait loci (QTL) and haplotype multiplicity of the genes concerned. A limited number of quantitative trait nucleotides responsible for QTL variation have been described, but an acceleration in their rate of discovery is expected with the adoption of linkage disequilibrium and candidate gene strategies for QTL fine mapping and cloning. Additional layers of regulatory variation have been studied that could also contribute to the molecular basis of quantitative genetics of crop traits. Despite this progress, the role of marker assisted selection in plant breeding will ultimately depend on the genetic model underlying quantitative variation. PMID- 12732325 TI - Nutritional enhancement of plants. AB - Plants can provide most of the nutrients required in the human diet; however, the major staple crops are often deficient in some of these nutrients. Thus, malnutrition, with respect to micronutrients like vitamin A, iron and zinc, affects >40% of the world's population. Advances in molecular biology are being exploited to produce crops enhanced in these key nutrients. Other nutritional targets include the modification of fatty acid composition and the enhancement of antioxidant levels, particularly carotenoids, such as lycopene, and flavonoids. However, the benefit of these 'biofortified' crops to human nutrition remains to be elucidated. PMID- 12732326 TI - Fungal genomics beyond Saccharomyces cerevisiae? AB - Fungi are used extensively in both fundamental research and industrial applications. Saccharomyces cerevisiae has been the model organism for fungal research for many years, particularly in functional genomics. However, considering the diversity within the fungal kingdom, it is obvious that the application of the existing methods of genome, transcriptome, proteome and metabolome analysis to other fungi has enormous potential, especially for the production of food and food ingredients. The developments in the past year demonstrate that we have only just started to exploit this potential. PMID- 12732328 TI - Exploitation of molecular profiling techniques for GM food safety assessment. AB - Several strategies have been developed to identify unintended alterations in the composition of genetically modified (GM) food crops that may occur as a result of the genetic modification process. These include comparative chemical analysis of single compounds in GM food crops and their conventional non-GM counterparts, and profiling methods such as DNA/RNA microarray technologies, proteomics and metabolite profiling. The potential of profiling methods is obvious, but further exploration of specificity, sensitivity and validation is needed. Moreover, the successful application of profiling techniques to the safety evaluation of GM foods will require linked databases to be built that contain information on variations in profiles associated with differences in developmental stages and environmental conditions. PMID- 12732327 TI - Metabolic pathway engineering in lactic acid bacteria. AB - Lactic acid bacteria (LAB) display a relatively simple carbon and energy metabolism where the sugar source is converted mainly to lactic acid. In Lactococcus lactis metabolic engineering has been very successful in the re routing of lactococcal pyruvate metabolism to products other than lactic acid. Current metabolic engineering approaches tend to focus on more complex, biosynthetic pathways leading to end-products that generate a health benefit for the consumer (nutraceuticals). Several examples of research on these minor pathways in L. lactis have illustrated the potential of LAB as producers of these metabolites. Whole genome sequencing efforts and corresponding global technologies will have an impact on metabolic engineering in the future. PMID- 12732329 TI - Control of antimicrobial peptide synthesis by the agr quorum sensing system in Staphylococcus epidermidis: activity of the lantibiotic epidermin is regulated at the level of precursor peptide processing. AB - The accessory gene regulator (agr) quorum sensing system in staphylococci is responsible for the regulation of surface proteins and exoproteins, including many virulence factors in the pathogenic species Staphylococcus aureus and S. epidermidis. Strain S. epidermidis Tu3298 produces the lantibiotic epidermin. An isogenic agr deletion mutant of this strain showed a strong reduction of epidermin production. Detailed analysis of the impact of agr on epidermin biosynthesis revealed that agr does not interfere with the transcription of epidermin biosynthetic genes, but controls the extracellular processing of the N terminal leader peptide by the EpiP protease. PMID- 12732330 TI - Peptides derived from the two regulatory domains of p53 are recognized by two p53 activating antibodies. AB - The C-terminus of the transcription factor p53 seems to play an important role by controlling the specific DNA-binding activity, which is directly associated with sensing damaged DNA. Another region located in the N-terminus of the protein has also been shown to regulate the DNA-binding activity of the protein. This activity can be promoted by peptides derived from these two negative regulatory regions or by binding of antibodies directed against the C-terminus of the p53 protein. Using both phage display peptide and multiple peptide synthesis technologies, we demonstrated that mAbs HR231 and Pab421, two p53-activating antibodies, recognize peptides derived from the C-terminus of p53, as previously described, but also peptides from the N-terminus of the protein, suggesting that these peptides are part of a conformational epitope. Furthermore, the sequences of these peptides are located in the two negative regulatory regions identified on the p53 protein, which is consistent with the biological activity of mAbs HR231 and Pab421. PMID- 12732331 TI - Analysis of amino acid pairs sensitive to variants in human collagen alpha5(IV) chain precursor by means of a random approach. AB - In this data-based theoretical analysis, we use the random approach to analyze the amino acid pairs in 5(IV) chain precursor (CA54) in order to determine which amino acid pairs are more sensitive to 151 variants from missense mutant human CA54 protein. The rationale of this study is based on our hypothesis and previous findings that harmful variance is more likely to occur at randomly unpredictable amino acid pair position rather than at randomly predictable positions. This is reasonable to argue as randomly predictable amino acid pairs are less likely to be deliberately evolved, whereas randomly unpredictable amino acid pairs are probably deliberately evolved in connection with protein function. The results show that all 151 variants occurred at randomly unpredictable amino acid pairs and the chance of a variant occurring is markedly higher in randomly unpredictable amino acid pairs than in predictable pairs. Thus, randomly unpredictable amino acid pairs are more sensitive to variance in human CA54. The results also suggest that the human CA54 protein has a natural tendency towards variants. PMID- 12732332 TI - Constructs of biotin mimetic peptide with CC49 single-chain Fv designed for tumor pretargeting. AB - Single-chain Fv constructs comprising a biotin mimetic peptide (BMP) and scFv of CC49 monoclonal antibody were produced to improve pretargeted radioimmunotherapy. BMP units that bind streptavidin were added to the carboxyl terminus of the CC49 V(H) region. An engineered scFvBMP monomer and a sc(Fv)(2)BMP dimer showed an excellent antigen recognition in vitro with a specific binding of 72+/-5 and 81+/ 4%, respectively. Properties of 125I-sc(Fv)(2)BMP in mice bearing LS-174T xenografts were comparable to these of the parent 125I-sc(Fv)(2). Complexing of scFvBMPs with streptavidin increased tumor targeting and gave exceptionally high tumor-to-blood values of 63+/-7 for 125I-sc(Fv)(2)BMP-streptavidin compared with 37+/-4 for sc(Fv)(2)BMP at 72h after administration. High tumor and negligible normal tissue levels of these novel pretargeting constructs indicate a great potential for pretargeted radioimmunotherapy. PMID- 12732333 TI - Circadian telomerase activity and DNA synthesis for timing peptide administration. AB - DNA synthesis and telomerase activity were assessed in nude mice transplanted with hepatic carcinoma. Hepatic cancer cells (SMMC-7721) were implanted into both flanks of each of 14 BALB/C mice synchronized in 12 h of light alternating with 12 h of darkness (LD12:12) for 4 weeks. At 7 timepoints, tumor samples were collected for measurement of cellular DNA content by flow cytometry and telomerase activity by PCR-ELISA assay. Cosinor analyses determine a 24-h rhythm for all variables, showing a similar timing for the DNA-synthesis phase and telomerase activity. These results provide a model for exploring optimal timing of chronotherapy with peptides, especially for treatment with telomerase inhibitors. PMID- 12732334 TI - Degradation of the immunogenic peptide gp100(280-288) by the monocyte-like U937 cell line. AB - The possible degradation of the tumor antigen epitope gp100(280-288) (YLEPGPVTA) in the presence of the monocyte-like line U937, and the effect of degradation on the in vitro-measured immune recognition, were investigated by chromatographic techniques and immunological assays. Results indicate a rapid hydrolysis of the substrate in the presence of the model cells, which is consistent with the hypothesis that degradation of gp100(280-288) is caused by the activity of U937 expressed enzymes, specifically amino- and carboxypeptidases. On the other hand, these results do not support the involvement of other enzymes known to be expressed by U937 cells. From a functional point of view, these data indicate that the degradation of gp100(280-288) severely hampered recognition by specific CTL clones. The results obtained may provide a model for epitope degradation by the antigen-presenting cells found in defined anatomical compartments and may, at least in part, account for the low activity of peptide-based antineoplastic vaccines, as well as for the transience of the effects of subcutaneously administered peptides in general. PMID- 12732335 TI - Peptidomic analysis of the skin secretions of the pickerel frog Rana palustris identifies six novel families of structurally-related peptides. AB - Peptidomics methodology has been used to identify and characterize structurally 26 previously undescribed peptides in the electrically stimulated skin secretions of the North American pickerel frog Rana palustris. Peptides in the secretions were analyzed by electrospray mass spectrometry and components in mass range 700 2500 Da, present in major abundance, were purified by reverse-phase HPLC. Cysteine-containing components were identified by treatment with dithiothreitol and 4-vinylpyridine and re-analysis of the derivatizated peptide by mass spectrometry. Application of these techniques led to the identification of six families of structurally-related peptides comprising (a). six peptides containing the consensus sequence Cys-Trp-Xaa-Thr-Lys-Ser-Ile-Pro-Pro-Lys/Arg-Xaa-Cys, (b). three peptides containing the consensus sequence Pro-Pro-Gly-Val-Cys-(Xaa)(3) Lys/Arg-Arg-Cys, (c). two peptides containing the consensus sequence Ser-Phe-His Val-Phe-Pro-Pro-Trp-Met-Cys-Lys-Xaa-Leu-Lys-Lys-Cys, (d). two peptides containing the consensus sequence Arg-Xaa-Cys-Trp-Lys-(Xaa)(2)-Asn-(Xaa)(3)-Val-Cys-Ser, (e). nine peptides containing the consensus sequence Ser-Leu-Pro-Ala-Gly-Leu-Ser Pro, and (f). four peptides containing the consensus sequence Asp-Xaa-Gln-Asp-Arg Trp-Xaa-Pro. The peptides did not inhibit the growth of Escherichia coli or Staphylococcus aureus and were inactive on hamster vascular or gastric smooth muscle preparations so that their biological functions, if any, remain to be established. PMID- 12732336 TI - Cloning, pharmacology, and distribution of the neuropeptide Y-receptor Yb in rainbow trout. AB - This work describes the isolation and pharmacological characterization of a neuropeptide Y (NPY) receptor from rainbow trout (Oncorhynchus mykiss). The receptor exhibits approximately 45% amino acid sequence identity to mammalian Y1 subfamily receptors, Y1, Y4 and y6, a similar degree of identity as these subtypes display to one another. Because it displays highest sequence identity to zebrafish Yb (75%), we named it the trout Yb receptor. The receptor exhibits high binding affinity for zebrafish and human NPY and peptide YY (PYY) but not truncated forms of the peptides. Human pancreatic polypeptide (PP) also binds with high affinity. Y1 selective antagonists exhibit poor binding as is the case for Y2 and Y5 selective ligands. This binding profile supports membership in the Y1 subfamily. Sequence data also support this relationship suggesting that Yb is a fourth and separate member of the Y1 subfamily. NPY has a number of important physiological functions such as regulating food intake and reproduction. The expression of the receptor in the hypothalamus and telencephalon suggests a possible role in these processes. This and other receptors from this species have potential for improving aquaculture. PMID- 12732337 TI - beta-MSH: a functional ligand that regulated energy homeostasis via hypothalamic MC4-R? AB - alpha-Melanocyte stimulating hormone (MSH) has generally been assumed to be the endogenous ligand acting at the melanocortin-4 receptor (MC4-R), activation of which in the hypothalamus leads to reduced feeding. However, beta-MSH is also capable of activating MC4-R and inhibiting feeding. Here, we investigated the possibility that beta-MSH acts as an endogenous MC4-R agonist and that this melanocortin peptide plays a role in the regulation of feeding and energy balance. We found that beta-MSH had significantly higher affinities than alpha MSH at both human MC4-R transfected into CHO cells (K(i): beta-MSH, 11.4+/-0.4 nmol/l versus alpha-MSH, 324+/-16 nmol/l, P<0.001) and MC4-R in rat hypothalamic homogenates (K(i): beta-MSH, 5.0+/-0.4 nmol/l versus alpha-MSH, 22.5+/-2.3 nmol/l, P<0.001). Incubation of brain slices with 5 microM beta-MSH significantly increased [35S]GTPgammaS binding by 140-160% (P<0.001), indicating activation of G-protein-coupled receptors (GPCRs), in the hypothalamic ventromedial (VMH), dorsomedial (DMH), arcuate (ARC) and paraventricular (PVN) nuclei. These sites match the distribution of beta-MSH immunoreactive fibres and also the distribution of MC4-R binding sites which we and others previously reported. Food restriction significantly increased beta-MSH levels in the VMH, DMH and ARC (all P<0.05) above freely-fed controls, whilst alpha-MSH concentrations were unchanged. We propose that increased beta-MSH concentrations reflect blockade of the peptide's release in these sites, consistent with the increased hunger and the known up-regulation of MC4-R in the same nuclei. Thus, we conclude that (1). beta-MSH has higher affinity at MC4-R than alpha-MSH; (2). beta-MSH activates GPCR in these sites, which are rich in MC4-R; and (3). beta-MSH is present in hypothalamic nuclei that regulate feeding and its concentrations alter with nutritional state. We suggest that beta-MSH rather than alpha-MSH is the key ligand at the MC4-R populations that regulate feeding, and that inhibition of tonic release of beta-MSH is one mechanism contributing to hunger in under feeding. PMID- 12732338 TI - Plasma orexin-A-like immunoreactivity in patients with sleep apnea hypopnea syndrome. AB - Orexin-A (hypocretin-1), a neuropeptide produced in hypothalamus, stimulates arousal. We studied plasma concentrations of orexin-A-like immunoreactivity (orexin-A-LI) in 156 patients with sleep apnea hypopnea syndrome (SAHS) and 22 control subjects. Plasma orexin-A-LI levels were significantly decreased in 156 patients with SAHS (4.4+/-0.15 pmol/l, mean+/-S.E.) as compared with controls (5.3+/-0.45 pmol/l). The levels were decreased in parallel with the severity of sleep-related respiratory disturbance and magnitude of sleep fragmentation. These findings raise the possibility that a low plasma level of orexin-A-LI may be a marker to show the severity of the disease in patients with SAHS. PMID- 12732339 TI - Endogenous CART peptide regulates mu opioid and serotonin 5-HT(2A) receptors. AB - Previous experiments conducted in this laboratory showed that intracerebroventricular (i.c.v.) administration of IgG antibodies directed against selected neuropeptides changed the density of CNS receptors, suggesting that neuropeptides in the cerebrospinal fluid can perform a regulatory role. To further test this hypothesis, we administered anti-CART peptide (the peptide product of cocaine amphetamine related transcript) IgG to rats via the i.c.v. route, and measured the density of opioid mu and delta receptors, beta-adrenergic and alpha(2)-adrenergic receptors and serotonin 5-HT(2A) receptors using ligand binding methods. We also used Western blots to determine the expression level of the mu, delta and 5-HT(2A) receptors. The results demonstrated that anti-CART peptide IgG up-regulates mu and 5-HT(2A) receptor in the hippocampus and caudate We conclude that CART peptides in the cerebrospinal fluid may exert regulatory effects in the brain. PMID- 12732340 TI - In vitro and in vivo opioid activity of [DPro(6)]dermorphin, a new dermorphin analogue. AB - To study the effects of inducing stereo-chemical modifications in the structure of dermorphin (DM) so as to improve its mu-opioid receptor affinity and its resistance to C-terminal enzymatic degradation, in the Institute of Molecular Genetics of Moscow, we synthesized a new DM analogue ([DPro(6)]DM) and analyzed the changes induced in the biological activities of DM by substituting the Pro(6) residue with DPro(6). We compared the activity of the new DM analogue and DM in in vitro assays and in in vivo tests of analgesia, thermoregulation, heart rate recordings, and gastrointestinal motility in rats. In the in vitro tests, guinea pig ileum (GPI) and mouse vas deferens (MVD), although the opioid activities of [DPro(6)]DM indicated that the peptide was always less potent than DM, its lower IC(50) ratios (mu/delta) showed that it had higher mu-opioid receptor selectivity. In the in vivo analgesic test, [DPro(6)]DM, when injected intraperitoneally (i.p.) (0.5-5 and 10mg/kg) in rats, had the same antinociceptive efficacy as DM and when injected intranasally (i.n.) (0.005 and 0.02 mg/kg) it induced a more stable and long-lasting analgesia than DM (the AUC was about 91% higher for [DPro(6)]DM than for DM). Moreover, these data confirm that the intranasal route is advantageous for peripheral drug administration. In the heart rate study, [DPro(6)]DM and DM (0.5mg/kg, i.p.), induced a similar, weak bradycardia. The only difference was that [DPro(6)]DM induced a longer lasting effect than DM. Conversely, in body temperature regulation [DPro(6)]DM induced weaker inhibitory activity than DM (56% of the DM-induced response); it did so only in a cold environment and at the maximal used dose (0.5mg/kg, i.p.) without inducing vasomotor effects. In the gastrointestinal study, [DPro(6)]DM and DM (0.005, 0.05, and 0.5mg/kg, i.p.) significantly slowed upper gastrointestinal transit of a charcoal meal and inhibited colonic propulsion. Comparison of the ED(50) values of [DPro(6)]DM (0.03 mg/kg) and DM (0.009 mg/kg) showed that the DM analogue was about three times less potent than DM in slowing gastrointestinal and colonic transit. In conclusion, all these data overall suggest that structural maneuvering in the Pro(6)-residue of the DM molecule changes its affinity for mu-opioid receptor subtypes and confirms the usefulness of experimental studies involving structural modifications in obtaining new therapeutic agents. PMID- 12732341 TI - VIP and PACAP receptors coupled to adenylyl cyclase in human lung cancer: a study in biopsy specimens. AB - Vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP) are important neuropeptides in the control of lung physiology. Both of these commonly bind to specific G protein coupled receptors named VPAC(1)-R and VPAC(2)-R, and PAC(1)-R (with higher affinity for PACAP). VIP and PACAP have been implicated in the control of cell proliferation and tumor growth. This study examined the presence of VIP and PACAP receptors in human lung cancer samples, as well as the functionality of adenylyl cyclase (AC) stimulated by both peptides. Results from RT-PCR and immunoblot experiments showed the expression of VPAC(1)-, VPAC(2)- and PAC(1)-R in lung cancer samples. Immunohistochemical studies showed the expression of VPAC(1) and VPAC(2) receptors. These receptors were positively coupled to AC, but the enzyme activity was impaired as compared to normal lung. There were no changes in Galpha(s) or Galpha(i) levels. Present results contribute to a better knowledge of VIP/PACAP actions in lung cancer and support the interest for the development of VIP/PACAP analogues with therapeutic roles. PMID- 12732342 TI - Passage of vasoactive intestinal peptide across the blood-brain barrier. AB - We investigated the ability of vasoactive intestinal peptide (VIP) to cross the blood-brain barrier (BBB), the interface between the peripheral circulation and central nervous system (CNS). VIP labeled with 131I (I-VIP) and injected intravenously into mice was taken up by brain as determined by multiple-time regression analysis. Excess unlabeled VIP was unable to impede the entry of I VIP, indicating that passage is by nonsaturable transmembrane diffusion. High pressure liquid chromatography (HPLC) showed the radioactivity entering the brain to be intact I-VIP. After intracerebroventricular (i.c.v.) injection, I-VIP was sequestered by brain, slowing its efflux from the CNS. In summary, VIP crosses the BBB unidirectionally from blood to brain by transmembrane diffusion. PMID- 12732343 TI - Cyclo (His-Pro) potentiates GABA/ethanol-mediated chloride uptake by neurosynaptosomes. AB - Cyclo (His-Pro) CHP is a cyclic dipeptide endogenous to the brain of a variety of animal species including man. Administration of exogenous peptide to rodents has been shown to exhibit a variety of biologic activities including, modification of pharmacologic actions of alcohol. Since there are many apparent similarities between the actions of GABA and CHP in modulating alcohol pharmacology, we have examined whether CHP can modulate alcohol potentiation of GABA-receptor-mediated 36Cl-influx in neurosynaptosomes. The results show a further dose-dependent potentiation of 36Cl-influx in neurosynaptosomes by CHP in the presence of GABA and alcohol. PMID- 12732344 TI - Elevated glucose blocks angiotensin-(1-7) and bradykinin interaction: the role of cyclooxygenase products. AB - The interaction between angiotensin-(1-7) [Ang-(1-7)] and bradykinin (BK) was studied in the isolated mesenteric arteriolar bed of control and diabetic rats perfused with either 5.5 or 22 mM of glucose. Prostanoids release after the administration of BK, Ang-(1-7) and Ang-(1-7)+BK was also studied. In control and diabetic preparations perfused with Krebs Henseleit solution with 5.5mM of glucose, Ang-(1-7) potentiates BK-induced vasodilation. On the other hand, the potentiating effect disappeared in control and diabetic preparations perfused with 22 mM of glucose. Prostaglandin F(2alpha) (PGF(2alpha)) release induced by BK and Ang-(1-7)+BK was increased in perfusates of diabetic preparations containing 22 mM of glucose. The release of thromboxane A(2) (TXA(2)) (measured as TXB(2)) or prostaglandin I(2) (PGI(2)) (measured as 6-keto-PGF(1alpha)) did not differ in control and diabetic preparations perfused with 5.5 and 22 mM of glucose. Our data allow us to suggest that hyperglycemia may be involved in the lack of potentiation in control and diabetic preparations; increase in PGF(2alpha) release, but not other cyclooxygenase products, may explain the absence of potentiation in diabetic preparations. PMID- 12732345 TI - Characterization of angiotensin-(1-7) receptor subtype in mesenteric arteries. AB - Mesenteric arteries from male Sprague-Dawley rats were mounted in a pressurized myograph system. Ang-(1-7) concentration-dependent responses were determined in arteries preconstricted with endothelin-1 (10(-7)M). The receptor(s) mediating the Ang-(1-7) evoked dilation were investigated by pretreating the mesenteric arteries with specific antagonists of Ang-(1-7), AT(1) or AT(2) receptors. The effects of Ang-(3-8) and Ang-(3-7) were also determined. Ang-(1-7) caused a concentration-dependent dilation (EC(50): 0.95 nM) that was blocked by the selective Ang-(1-7) receptor antagonist D-[Ala(7)]-Ang-(1-7). Administration of a specific antagonist to the AT(2) receptor (PD123319) had no effect. On the other hand, losartan and CV-11974 attenuated the Ang-(1-7) effect. These results demonstrate that Ang-(1-7) elicits potent dilation of mesenteric resistance vessels mediated by a D-[Ala(7)]-Ang-(1-7) sensitive site that is also sensitive to losartan and CV-11974. PMID- 12732347 TI - Tachykinin peptide-induced activation and desensitization of neurokinin 1 receptors. AB - The actions of four tachykinins on inhibition and desensitization of the M current of bullfrog sympathetic neurons have been characterized. Radioligand binding parameters of the tachykinins were determined at a neurokinin receptor in a heterologous expression system. The correlation between binding, signaling and receptor regulation was investigated. A correlation between receptor binding and signaling was found between the peptides; however, their ability to produce desensitization was not correlated with binding and signaling. These results show that the ability of a tachykinin peptide to induce signal activation is not indicative of its ability to induce receptor regulation. PMID- 12732346 TI - Changes of adrenomedullin and receptor activity modifying protein 2 (RAMP2) in myocardium and aorta in rats with isoproterenol-induced myocardial ischemia. AB - Adrenomedullin is a potent vasodilator peptide originally isolated from a pheochromocytoma. Recently, a novel adrenomedullin receptor has been identified as a complex of calcitonin receptor-like receptor (CRLR) and receptor activity modifying protein 2 (RAMP2). To explore the pathophysiological roles of adrenomedullin and its receptor component RAMP2 in ischemic cardiovascular diseases, we studied the changes of adrenomedullin and RAMP2 mRNA in myocardium and aorta in rats with isoproterenol (ISO)-induced myocardial impairment. In ISO treated rats, heart became enlarged markedly, the ratio of heart to body weight was increased by 54% (P<0.01), and myocardial malondialdehyde content and plasma lactate dehydrogenase activity was elevated by 43% (P<0.01) and 138% (P<0.01), respectively. Immunoreactive adrenomedullin (ADM) in plasma, myocardium and aorta was augmented by 116.7% (P<0.01), 50.8% (P<0.01) and 12.5% (P>0.05), respectively. ADM mRNA in myocardium and aorta was increased by 96.8% (P<0.01) and 38.5% (P<0.01), respectively. RAMP2 mRNA in myocardium and aorta was increased by 19.6% (P<0.05) and 15.8% (P<0.01), respectively. These results suggest that the increase of ADM level and the up-regulation of ADM and RAMP2 gene in myocardium and aorta may be significant in the pathogenesis of ischemic myocardiopathy. PMID- 12732348 TI - Endomorphin-1 and -2 induce naloxone-precipitated withdrawal syndromes in rats. AB - In 1997, endomorphin-1 (EM-1) and -2 (EM-2) were identified as the most specific endogenous mu-opioid ligands. These two peptides have shown analgesic effects and many other opioid functions. In the present study, we attempt to investigate the possible ability of endomorphins to induce naloxone-precipitated withdrawal in comparison with that induced by morphine. Using the previously established scoring system in rats, 12 withdrawal signs (chewing, sniffing, grooming, wet-dog shakes, stretching, yawning, rearing, jumping, teeth grinding, ptosis, diarrhea, and penile erection) were observed and scored following naloxone (4 mg/kg, i.p.) challenge. Compared with the sham control, EM-1 and EM-2 (20 microg, i.c.v., b.i.d. for 5 days) both produced significant naloxone-induced withdrawal syndromes with similar severity to that induced by the same dose of morphine. There was no significant difference between EM-1, EM-2, and morphine-treated group for naloxone-induced withdrawal signs, except for grooming. EM-1 and EM-2 induced more grooming than that caused by morphine. Although EM-1 and EM-2 both led to the withdrawal, they displayed different potency for certain signs and suggest their distinct regulations. The present results indicate EM-1 and EM-2 could initiate certain mechanism involved opiate dependence. PMID- 12732349 TI - Plasma motilin in untreated celiac disease. AB - Celiac disease (CD) is characterized by mucosal villous atrophy mostly confined to the proximal small intestine. Upper-gut motor abnormalities have been reported. Motilin, localized in cells in the proximal small intestine, is a trigger factor for the migrating motor complex. Plasma levels of motilin were studied in 16 untreated CD patients and in an age-matched control group of 18 healthy subjects by radioimmunoassay and by high-performance liquid chromatography (HPLC). The fasting levels of motilin and postprandial levels were significantly higher in CD patients compared to controls (P<0.01) and HPLC revealed a divergent individual pattern of the motilin fragments. PMID- 12732350 TI - Targeting ICAM-1/LFA-1 interaction for controlling autoimmune diseases: designing peptide and small molecule inhibitors. AB - This review describes the role of modulation of intracellular adhesion molecule-1 (ICAM-1)/leukocyte function-associated antigen-1 (LFA-1) interaction in controlling autoimmune diseases or inducing immunotolerance. ICAM-1/LFA-1 interaction is essential for T-cell activation as well as for migration of T cells to target tissues. This interaction also functions, along with Signal-1, as a co-stimulatory signal (Signal-2) for T-cell activation, which is delivered by the T-cell receptors (TCR)-major histocompatibility complex (MHC)-peptide complex. Therefore, blocking ICAM-1/LFA-1 interaction can suppress T-cell activation in autoimmune diseases and organ transplantation. Many types of inhibitors (i.e. antibodies, peptides, small molecules) have been developed to block ICAM-1/LFA-1 interactions, and some of these molecules have reached clinical trials. Peptides derived from ICAM-1 and LFA-1 sequences have been shown to inhibit T-cell adhesion and activation. In addition, these inhibitors have been useful in elucidating the mechanism of ICAM-1/LFA-1 interaction. Besides binding to LFA-1, the ICAM-1 peptide can be internalized by LFA-1 receptors into the cytoplasmic domain of T-cells. Therefore, this ICAM-1 peptide can be utilized to selectively target toxic drugs to T-cells, thus avoiding harmful side effects. Finally, bi-functional inhibitory peptide (BPI), which is made by conjugating the antigenic peptide and an LFA-1 peptide, can alter the T-cell commitment from T helper-1 (Th1) to T-helper-2 (Th2)-like cells, suggesting that this peptide may have a role in blocking the formation of the "immunological synapse." PMID- 12732351 TI - Modulatory effects of hormones, drugs, and toxic events on renal organic anion transport. AB - The human body is exposed continuously to a wide variety of exogenous compounds, many of which are anionic compounds. In addition, products of phase II biotransformation reactions are negatively charged, viz. glucuronides, sulfate esters, or glutathiones. Renal transport of organic anions is an important defense mechanism of the organism against foreign substances. The combination of the rate of uptake and efflux and the intracellular disposition of organic anions in the proximal tubule determines the intracellular concentration and the nephrotoxic potential of a compound. Modulation of organic anion secretion is observed after exposure of proximal tubules to various hormones, and the subsequent receptor-mediated response is signaled by protein kinases. Transport of anionic compounds across the basolateral as well as the luminal membrane is modified by activation or inhibition of protein kinases. Protein kinase C activation reduces the uptake of organic anions mediated by the organic anion transporter 1 (OAT1/Oat1) and Oat3 and reduces Mrp2-mediated efflux. In addition, activation of protein kinase C has been shown to inhibit transport by the organic anion transporting polypeptide 1 (Oatp1) across the luminal membrane. Additional protein kinases have been implicated in the regulation of organic anion transport, and the role of nuclear factors in xenobiotic excretion is an emerging field. The physiological regulation of organic anion transporters may also be influenced by exogenous factors, such as exposure to xenobiotics and cellular stress. This commentary discusses the current knowledge of endogenous and exogenous influences on renal anionic xenobiotic excretion. PMID- 12732352 TI - Discovery and characterization of a potent and selective non-amidine inhibitor of human factor Xa. AB - Benzothiophene-anthranilamide 1 (3-chloro-N-[2-[[(4-fluorophenyl)amino]carbonyl] 4-methylphenyl]benzo[b]thiophene-2-carboxamide) was discovered by high throughput screening to be a highly potent and selective non-amidine inhibitor of human factor Xa with a K(i) of 15+/-4nM. Compound 1 is a selective inhibitor of human factor Xa as suggested by the K(i)((app)) determined for nine other human serine proteases and bovine trypsin. The activity of reconstituted human prothrombinase complex was inhibited by compound 1 when assayed in physiological concentrations of the substrate prothrombin. However, 27-fold higher inhibitor concentrations were needed to achieve the same level of inhibition than were required for the inhibition of free factor Xa, due in part to non-specific binding of the inhibitor to phospholipid under the assay conditions. Failure to demonstrate enzymatic cleavage of compound 1 suggests that compound 1 is solely an inhibitor rather than a substrate for factor Xa. The inhibition of factor Xa by compound 1 was reversible upon dilution of the enzyme/inhibitor mixture. Analyses of the inhibition mechanism with Dixon, Cornish-Bowden, and Lineweaver-Burk plots showed that compound 1 is a linear mixed-type inhibitor with 5-fold higher affinity for free factor Xa than the factor Xa/substrate complex. The linear mixed-type inhibition suggests that compound 1 binds to the active site region of factor Xa, but its binding cannot be fully displaced by the substrate S2222 (1:1 mixture of N-benzoyl-Ile-Glu-Gly-Arg-p-nitroanilide and N-benzoyl-Ile-Glu(gamma-OMe)-Gly-Arg p-nitroanilide hydrochloride). Thus, the inhibition mechanism for compound 1 is novel compared to most serine protease inhibitors including amidine-containing factor Xa inhibitors, which rely on binding to the S1 pocket of the enzyme active site. Compound 1 represents an attractive, novel structural template for further development of efficacious, safe, and potentially orally active human factor Xa inhibitors. PMID- 12732353 TI - Multidrug resistance correlates with overexpression of Muc4 but inversely with P glycoprotein and multidrug resistance related protein in transfected human melanoma cells. AB - Due to the size, glycosylation, and location in the plasma membrane of the sialomucin complex Muc4, which has been implicated in ErbB2 signaling, in the repression of apoptosis and cell adhesion, and in tumor metastasis, studies were initiated to determine whether its presence could influence cell sensitivity to anticancer drugs. Growth inhibition assays using melanoma cell lines that either express the glycoprotein (Muc4(+)) or do not (Muc4(-)) showed that Muc4 renders cells resistant to taxol, doxorubicin, vinblastine, rhodamine 123, and 2 deoxyglucose. When treated with various concentrations of doxorubicin, Muc4(+) cells were blocked less frequently in G(2) and underwent less DNA fragmentation (apoptosis and/or necrosis) than Muc4(-) cells. All of the drugs tested (except for 2-deoxyglucose) are well recognized by P-glycoprotein-mediated multidrug resistance 1 (MDR1) and to a lesser degree by multidrug resistance related protein 1 (MRP1) transporters. Therefore, transporter gene expression in these cells was assayed. Surprisingly, Muc4(+) cells expressed lower levels of both transporter genes than Muc4(-) cells. Moreover, rhodamine 123 was retained more highly in the Muc4(+) than in the Muc4(-) cells, demonstrating that these transporters are functional. Overall, these results indicate that although Muc4(+) cells express less MDR1 and MRP1, they are more resistant to drugs recognized by these transporters. PMID- 12732354 TI - Further studies on the interaction of nonpolyglutamatable aminopterin analogs with dihydrofolate reductase and the reduced folate carrier as determinants of in vitro antitumor activity. AB - Thirteen structural analogs of the potent nonpolyglutamatable dihydrofolate reductase inhibitor N(alpha)-(4-amino-4-deoxypteroyl)-N(delta)-hemiphthaloyl-L ornithine (PT523) with modifications in the side chain, the para-aminobenzoyl moiety, or the 9,10-bridge were evaluated for the ability to inhibit human recombinant dihydrofolate reductase (DHFR), to utilize the reduced folate carrier (RFC) for influx, and to inhibit the growth of CCRF-CEM human leukemia cells in culture. In spectrophotometric assays of the kinetics of the reduction of dihydrofolate by DHFR in the presence of NADPH, these compounds had K(i) values ranging from 0.2 to 1.3pM, and thus were not greatly different in potency from the parent drug PT523. By comparison, the K(i) values of aminopterin (AMT), methotrexate (MTX), and 10-ethyl-10-deazaaminopterin (EDX) were 3.7, 4.8, and 11pM. In assays of competitive inhibition of [3H]MTX influx into CCRF-CEM cells, the K(i) values ranged from 0.21 to 7.3 micro M, as compared with 0.71, 5.4, and 1.1 micro M for PT523, AMT, and EDX. The K(t) for MTX was also re-analyzed and found to be 4.7 micro M, in better agreement with the literature than our previously reported value of 7.1 micro M. The IC(50) values of these compounds as inhibitors of the growth of CCRF-CEM cells after 72hr of drug exposure ranged from 0.53 to 55nM, and were qualitatively consistent with the other results. PMID- 12732355 TI - Molecular mechanisms for the activation of Ca2+-permeable nonselective cation channels by endothelin-1 in C6 glioma cells. AB - We recently demonstrated that endothelin-1 (ET-1) activates two types of Ca(2+) permeable nonselective cation channels (NSCC-1 and NSCC-2) in C6 glioma cells. It is possible to discriminate between these channels by using the Ca(2+) channel blockers SK&F 96365 (1-[beta-(3-[4-methoxyphenyl]propoxy)-4-methoxyphenethyl]-1H imidazole hydrochloride) and LOE 908 [(R,S)-(3,4-dihydro-6,7-dimethoxy isoquinoline-1-yl)-2-phenyl-N,N-di-[2-(2,3,4-trimethoxyphenyl)ethyl]-acetamide]. LOE 908 is a blocker for NSCC-1 and NSCC-2, whereas SK&F 96365 is an inhibitor for NSCC-2. The purpose of the present study was to identify the G-proteins that are involved in ET-1-activated Ca(2+) channels in C6 glioma cells. ET-1 activated only NSCC-1 in C6 glioma cells preincubated with U73122 (1-[6-[((17beta)-3 methoxyestra-1,3,5[10]-trien-17-yl)amino]hexyl]-1H-pyrrole-2,5-dione), a phospholipase C (PLC) inhibitor. Microinjection of the dominant negative mutant of G(12)/G(13) (G(12)G228A/G(13)G225A) abolished activation of NSCC-1 and NSCC-2. In contrast, pertussis toxin did not affect any of the Ca(2+) channels in the ET 1-stimulated C6 glioma cells. These results indicate that G(12)/G(13) may couple with endothelin receptors and play an important role in the activation of NSCCs in C6 glioma cells. Moreover, the activation mechanisms of NSCC-1 and NSCC-2 by ET-1 were different. NSCC-1 activation depended upon a G(12)/G(13)-dependent cascade, whereas NSCC-2 activation depended upon both G(q)/PLC- and G(12)/G(13) dependent cascades. PMID- 12732356 TI - The effect of valproic acid on drug and steroid glucuronidation by expressed human UDP-glucuronosyltransferases. AB - Valproic acid glucuronidation kinetics were carried our with three human UGT isoforms: UGT1A6, UGT1A9, and UGT2B7 as well as human liver and kidney microsomes. The glucuronidation of valproic acid was typified by high K(m) values with microsomes and expressed UGTs (2.3-5.2mM). The ability of valproic acid to interact with the glucuronidation of drugs, steroids and xenobiotics in vitro was investigated using the three UGT isoforms known to glucuronidate valproic acid. In addition to this the effect of valproic acid was investigated using two other UGT isoforms: UGT1A1 and UGT2B15 which do not glucuronidate valproic acid. Valproic acid inhibited UGT1A9 catalyzed propofol glucuronidation in an uncompetitive manner and UGT2B7 catalyzed AZT glucuronidation competitively (K(i)=1.6+/-0.06mM). Valproate significantly inhibited UGT2B15 catalyzed steroid and xenobiotic glucuronidation although valproate was not a substrate for this UGT isoform. No significant inhibition of UGT1A1 or UGT1A6 by valproic acid was observed. These data indicate that valproic acid inhibition of glucuronidation reactions is not always due to simple competitive inhibition of substrates. PMID- 12732357 TI - Induction of apoptosis in estrogen dependent and independent breast cancer cells by the marine terpenoid dehydrothyrsiferol. AB - Breast cancer (BCA) represents the highest incidence of death in 35- to 60-year old women. Above all, hormone unresponsive BCA is still associated with poorer prognosis than hormone receptor expressing malign, mammary tumors. There is a consistent need for effective compounds to treat especially the first variant of this disease. Therefore, we investigated the cytotoxic effects of the marine polyether triterpenoid dehydrothyrsiferol (DT) in four BCA cell lines. Annexin V labeling revealed higher rates of DT-induced apoptosis in hormone insensitive than in estrogen receptor expressing cells. Flow cytometric analysis of combined DNA fragmentation and total DNA labeling allowed us to ascribe apoptotic cells to their cell cycle stage. Although, high cell mortality was detected in mitogen dependent G(1)-phase, time, concentration, and cell line dependent populations of apoptotic cells were also found to be of S-phase and G(2)/M-phase origin. These results suggest that the induction of apoptosis by DT might be transduced through more than one effector pathway. Cell cycle distributions and 5-bromo-2' deoxyuridine incorporation varied in a treatment dependent manner and differed from control experiments with colchicine and doxorubicin which exclude that DT functions as a mitosis inhibitor. In summary, we propose that DT might be an interesting candidate for an antitumor drug development regimen. PMID- 12732358 TI - Important amino acids for the function of the human MT1 melatonin receptor. AB - Models of G protein-coupled melatonin receptor structure suggest that ligand recognition occurs in a binding pocket formed by transmembrane helices III, V and VII. Constitutively active mutations in G protein-coupled receptors have revealed that transmembrane helix III/intracellular loop 2 interface and transmembrane domain VI are critical regions in receptor activation. In this study, nine site directed mutants of the human MT1 melatonin receptor were created to test the importance of specific amino acids in these regions in ligand recognition and receptor activation events. We analyzed ligand binding, G protein activation and subcellular localization of MT1 receptors transiently expressed in COS-7 cells. Receptor ELISA was employed to study expression levels of N-terminally HA epitope tagged wild-type and mutant MT1 receptors. Mutations in histidine H195 (His(5.46)) in transmembrane domain V reduced receptor affinity for 2 [125I]iodomelatonin. Several other mutants had diminished expression on the plasma membrane. Amino acids M107 (Met(3.32)) in transmembrane domain III and S280 (Ser(7.46)) in transmembrane domain VII were found not to participate in ligand recognition in human MT1 receptor. Constitutive activity was not obtained with mutations in N124 (Asn(3.49)) or P253 (Pro(6.50)). These mutants failed to bind 2-[125I]iodomelatonin and had reduced expression levels. The need to upgrade current melatonin receptor models has become evident. Several important amino acids for the human MT1 melatonin receptor function were revealed in the current study, with effects of mutations ranging from slightly reduced affinity or efficacy to complete loss of function. PMID- 12732360 TI - Beneficial effects of L-carnitine in myoblastic C2C12 cells. Interaction with zidovudine. AB - L-Carnitine is a key molecule in the transfer of fatty acid across mitochondrial membranes. Bioavailable L-carnitine is either provided by an endogeneous biosynthesis or after intestinal absorption of dietary items containing L carnitine. After intestinal absorption or hepatic biosynthesis, L-carnitine is transferred to organs whose metabolism is dependent upon fatty acid oxidation, such as skeletal muscle. To cross the muscle plasma membrane, there are several transporters involved. Among those transporters, OCTN2 is actually the only one to have been clearly characterized. Zidovudine is a commonly used inhibitor of human immunodeficiency virus (HIV) replication. Zidovudine has many side effects, including induction of myopathy characterized by a metabolic mitochondria dysfunction and a diminution of the muscle L-carnitine content. In this study, we described the characteristics of L-carnitine transport in C2C12 cells. We also demonstrated that zidovudine inhibited the L-carnitine transporter. This inhibition led to a significant reduction of the muscle cell growth. In C2C12 cells, the supplementation of L-carnitine prevented the effects of zidovudine and restored the normal cell growth. PMID- 12732359 TI - Novel selective and metabolically stable inhibitors of anandamide cellular uptake. AB - Novel aromatic analogues of N-oleoylethanolamine and N-arachidonoylethanolamine (anandamide, AEA) were synthesized and, based on the capability of similar compounds to interact with proteins of the endocannabinoid and endovanilloid signaling systems, were tested on: (i) cannabinoid CB(1) and CB(2) receptors; (ii) vanilloid VR1 receptors; (iii) anandamide cellular uptake (ACU); and (iv) the fatty acid amide hydrolase (FAAH). The (R)- and, particularly, the (S)-1'-(4 hydroxybenzyl) derivatives of N-oleoylethanolamine and AEA (OMDM-1, OMDM-2, OMDM 3, and OMDM-4) inhibited to a varied extent ACU in RBL-2H3 cells (K(i) ranging between 2.4 and 17.7 micro M), the oleoyl analogues (OMDM-1 and OMDM-2, K(i) 2.4 and 3.0 micro M, respectively) being 6- to 7-fold more potent than the arachidonoyl analogues (OMDM-3 and OMDM-4). These four compounds exhibited: (i) poor affinity for either CB(1) (K(i)> or = 5 micro M) or CB(2) (K(i)>10 micro M) receptors in rat brain and spleen membranes, respectively; (ii) almost no activity at vanilloid receptors in the intracellular calcium assay carried out with intact cells over-expressing the human VR1 (EC(50)> or = 10 micro M); and (iii) no activity as inhibitors of FAAH in N18TG2 cell membranes (K(i)>50 micro M). The oleoyl- and arachidonoyl-N'-(4-hydroxy-3-methoxybenzyl)hydrazines (OMDM-5 and OMDM-6), inhibited ACU (K(i) 4.8 and 7.0 micro M, respectively), and were more potent as VR1 agonists (EC(50) 75 and 50nM, respectively), weakly active as CB(1) receptor ligands (K(i) 4.9 and 3.2 micro M, respectively), and inactive as CB(2) ligands (K(i)>5 micro M) as well as on FAAH (K(i)> or = 40 micro M). In conclusion, we report two novel potent and selective inhibitors of ACU (OMDM-1 and OMDM-2) and one "hybrid" agonist of CB(1) and VR1 receptors (OMDM-6). Unlike other compounds of the same type, OMDM-1, OMDM-2, and OMDM-6 were very stable to enzymatic hydrolysis by rat brain homogenates. PMID- 12732361 TI - Evaluation of beta-adrenergic receptor subtypes in the human prostate cancer cell line-LNCaP. AB - The present study was undertaken to determine the effects of catecholamines, agonists, and antagonists of beta-adrenergic receptors (AR) in the LNCaP cell line. Changes in cellular cyclic adenosine-3',5'-monophosphate (cAMP) levels were quantified by the use of a 6 cAMP response element (CRE)-luciferase reporter gene assay. LNCaP cells were transiently transfected with this gene construct, incubated in 96-well microtiter plates for 24 hr, and then treated with beta-AR agonists and/or antagonists for 4 hr. The rank order of potency for catecholamines and known beta-AR agonists was terbutaline(3.31 nM)>isoproterenol(8.31 nM)> or =fenoterol(15 nM)=epinephrine(16.2 nM)>norepinephrine(77.5 nM)>BRL-37344 [(R(*),R(*))-(+/-)4-[2-[(2-(3-chlorophenyl) 2-hydroxyethyl)amino]propyl]phenoxy acetic acid, sodium salt] (1000 nM)>dobutamine(1770 nM)>CGP12177 (4-[3-[(1,1-dimethylethyl)amino]-2 hydroxypropoxy]-1,3-dihydro-2H-benzimidazole-2-one hydrochloride) (inactive). The non-selective beta(1)-/-beta(2)-AR antagonists; propranolol and CGP 12177, at 10( 7)M, inhibited luciferase activity induced by these agonists by 80-96%. Propranolol blocked isoproterenol-induced luciferase responses in a competitive manner (K(B)=1.4 nM). In addition, isoproterenol-activated luciferase expression was blocked more potently by ICI 118,551 [(+/-)-1-[2,3-(dihydro-7-methyl-1H-inden 4-yl)oxy]-3-[(1-methylethy) amino]-2-butanol], a beta(2)-AR antagonist than by ICI 89,406 [(+/-)-N-[2-[3-(2-cyanophenoxy-)]-2-hydroxypropylamino]ethyl-N phenylurea], a beta(1)-AR antagonist, giving K(B) values of 1.07 and 161nM, respectively. These results suggest that the beta(2)-AR is the major subtype mediating catecholamine-induced cAMP changes in LNCaP cells. PMID- 12732362 TI - Effect of in vivo administered hexachlorobenzene on epidermal growth factor receptor levels, protein tyrosine kinase activity, and phosphotyrosine content in rat liver. AB - In the present study, the effects of hexachlorobenzene (HCB) on epidermal growth factor receptor (EGFR) content of liver microsomes and plasma membrane, and on EGFR-tyrosine kinase activity in the microsomal fraction were investigated. In addition, we studied the parameters of the tyrosine kinase signalling pathway such as protein tyrosine kinase (PTK) activity and phosphotyrosine content in microsomal and cytosolic protein. To determine whether the observed alterations were correlated with a manifestation of overt toxicity, a single very low dose of HCB (1mg/kg body wt) and two much higher doses (100 and 1000 mg/kg body wt), the highest being toxicologically significant in that it reduced serum thyroxine (T(4)) and inhibited uroporphyrinogen decarboxylase (URO-D) (EC 4.1.1.37) activity, were tested. Our results demonstrated that liver microsomes of rats treated with HCB had higher levels of EGFR than untreated rats; treated rats also had less EGFR present in hepatocyte plasma membrane fractions than did untreated rats. HCB altered the phosphotyrosine content and protein phosphorylation of some microsomal and cytosolic proteins in a biphasic dose-response relationship. At the low dose, phosphorylation and phosphotyrosine content of several microsomal proteins were increased; however, these effects were diminished or reversed at the higher doses. Our results suggest that chronic HCB treatment produces a down regulation of the EGFR and a dose-dependent increase in EGFR-tyrosine kinase activity in the microsomal fraction. This effect may contribute to the alteration of membrane and cytosolic protein tyrosine phosphorylation. The level of sensitivity encountered in our studies is extraordinary, occurring at 1/10 to 1/1000 the doses of HCB known to cause other toxicological lesions. PMID- 12732363 TI - Role of nerve growth factor in the regulation of parotid cell differentiation induced by rat serum. AB - The present study was undertaken to examine the factors that regulate rat serum (RS)- and nerve growth factor (NGF)-induced differentiation in a rat parotid acinar cell line. RS elicited extracellular signal-regulated kinase (ERK1/ERK2) activation within 5min, while cyclic AMP (cAMP) levels transiently rose after 6hr. RS also elicited a rise in amylase mRNA levels within 30min, which preceded the rise in amylase protein levels. A possible role for NGF was suggested by the findings that parotid cells express both TrkA and p75 receptors. The immunoreactivity of these NGF receptors was reduced during exposure to RS. Following prolonged incubation in RS when ERK activity subsided to near basal levels, NGF restored ERK1/ERK2 activity to the elevated level initially observed in RS. NGF was ineffective when cells were incubated in fetal bovine serum. NGF, when incubated in combination with the cAMP-generating neuropeptides, calcitonin gene-related peptide and vasoactive intestinal peptide, markedly enhanced the cellular amylase content produced by RS. We conclude that parotid cell differentiation arises from an activation of cell surface receptors by humoral factors in combination with NGF and cAMP-generating neuropeptides. PMID- 12732364 TI - Established theory of radiation-induced decay is not generalizable to Bolton Hunter labeled peptides. AB - Peptide hormones radiolabeled with 125I are widely used for the pharmacological characterization of cognate receptors. As a prerequisite for calculating ligand affinities from competition binding assays, and for estimating receptor densities from such studies, it is necessary to know the concentration of bioactive radioligand that is used in respective experiments. It has been demonstrated previously that radioiodinated peptides undergo decay catastrophe, i.e. disintegration of the radioactive label leads to the concomitant destruction of the carrier peptide. Here, we demonstrate that decay catastrophe does not apply to two peptide hormones that are iodinated by Bolton-Hunter conjugation: cholecystokinin octapeptide and glucagon-like peptide 2. The function of aged samples of these radioligands at corresponding recombinantly expressed receptors was assessed by measuring ligand-induced inositol phosphate production or generation of cyclic AMP, respectively. Both of the tested compounds, although predicted by decay catastrophe to contain little or subthreshold remaining bioactivity, stimulated an unexpectedly high level of receptor-mediated second messenger signaling. Quantitative comparison of observed functions with those of corresponding unlabeled peptides suggested that the bioactivity of each radioligand had been largely conserved despite the radioactive decay of the iodine label. Consistent with an apparent absence of decay catastrophe, we noted that the specific radioactivity, when determined immediately following peptide iodination, was close to the theoretical maximum but exponentially decreased over time. These findings raise the possibility that attachment of a Bolton-Hunter conjugate may shield labeled peptides from radiation-induced damage, a scenario that should be considered when performing radioligand binding experiments. PMID- 12732365 TI - Frequent co-occurrence of the TATA box mutation associated with Gilbert's syndrome (UGT1A1*28) with other polymorphisms of the UDP-glucuronosyltransferase 1 locus (UGT1A6*2 and UGT1A7*3) in Caucasians and Egyptians. AB - Polymorphisms of drug metabolizing enzymes are frequently associated with diseases and side effects of drugs. Recently, a TATA box mutation of UGT1A1 (UGT1A1*28), a common genotype leading to Gilbert's syndrome, and several missense mutations of other UDP-glucuronosyltransferase 1 (UGT1) family members have been described. Furthermore, co-occurrence of UGT1A1*28 and UGT1A6*2 has been observed. In order to elucidate the basis for co-occurrence of UGT1 mutations, fluorescence resonance energy transfer techniques were developed for rapid determination of polymorphisms of three UGT isoforms (UGT1A1*28, 1A6*2, and 1A7*2/*3). Hundred healthy Caucasians and 50 Egyptians were genotyped. All genotypes followed the Hardy-Weinberg equilibrium. Only three major haplotypes were found, including a haplotype consisting of allelic variants of all three isoforms (29% in Caucasians and 22% in Egyptians), all leading to reduced UGT activity. Frequent haplotypes containing several UGT1 allelic variants should be taken into account in studies on the association between diseases, abnormal drug reactions, and UGT1 family polymorphisms. PMID- 12732366 TI - Rabbit chronic ileitis leads to up-regulation of adenosine A1/A3 gene products, oxidative stress, and immune modulation. AB - A rabbit model of chronic ileitis has helped decipher the mechanism of alteration of multiple electrolyte and nutrient malabsorptions in inflammatory bowel disease (IBD). This study examined alterations in the adenosine A1/A3 receptor, oxidant, antioxidant, and immune-inflammatory pathways in chronic ileitis. Chronic ileal inflammation was induced 13-15 days after infection with 10,000 Eimeria magna oocytes. Quantitative analysis in 16 rabbits was done for oxidants, antioxidants, A1 and A3 transcripts, transport, injury, and inflammatory mediators. Inflamed gut had villus blunting, crypt hyperplasia and fusion, and immune cell infiltration. Alkaline phosphatase and Na-glucose co-transport were reduced by 78% (P=0.001) and 89% (P=0.001), respectively. Real-time fluorescence monitoring (TaqMan)-polymerase chain reaction revealed a transcriptional up-regulation of 1.34-fold for A1 and 5.40-fold for A3 receptors in inflamed gut. Lipid peroxidation increased in the mucosa (78%, P=0.012), longitudinal muscle myenteric plexus (118%, P=0.042), and plasma (104%, P=0.001). Mucosal antioxidants were altered by inflammation: reductions occurred in superoxide dismutase (32%, P=0.001) and catalase (43%, P=0.001), whereas increases occurred in glutathione (75%, P=0.0271) and glutathione reductase (86%, P=0.0007). Oxidant enzyme activities were elevated by 21% for xanthine oxidase (P=0.004), 172% for chloramine (P=0.022), 47% for gelatinase (P=0.041), and 190% for myeloperoxidase (P=0.002). Mast cell tryptase increased by 79% (P=0.006). Increases occurred in the plasma concentration of leukotriene B(4) (13-fold, P=0.003), thromboxane B(2) (61-fold, P=0.018), and tumor necrosis factor-alpha (9-fold, P=0.002). In conclusion, chronic ileitis and tissue injury are associated with discrete alterations in complex multi-level oxidant, antioxidant, and immune inflammatory components. The rabbit ileitis model is a suitable model to gain further insight into chronic inflammation and IBD. We hypothesize that adenosine A3 and A1 receptors may provide a novel target for therapy in chronic ileitis and perhaps IBD. PMID- 12732367 TI - LAAE-14, a new in vitro inhibitor of intracellular calcium mobilization, modulates acute and chronic inflammation. AB - A new lipidic acid-amido ether derivative (LAAE-14) able to reduce dose dependently the calcium increases mediated either by calcium ionophore ionomycin, by the endoplasmic reticular Ca(2+)-ATPase inhibitor thapsigargin, or by the chemotactic tripeptide N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP), in human neutrophils as well as in murine peritoneal macrophages, but not ATP, has been evaluated as a potential anti-inflammatory drug. This compound attenuated leukocyte activation by means of its inhibitory effect on the respiratory burst elicited in both types of cells by 12-O-tetradecanoyl phorbol 13-acetate, by inhibition of the degranulation process induced by cytochalasin B+fMLP or cytochalasin B+platelet activating factor, as well as by reduction of leukotriene B(4) synthesis induced by the calcium ionophore A23187. In addition, in zymosan stimulated mouse peritoneal macrophages LAAE-14 caused a potent inhibition of nitrite and prostaglandin E(2) production. This compound exerted acute and chronic anti-inflammatory effects by oral route, that may be related with several mechanisms such as attenuation of leukocyte activation, inhibition of inducible nitric oxide synthase, cyclo-oxygenase-2 and cytosolic phospholipase A(2) expression as well as reduction in tumour necrosis factor-alpha production. Its anti-inflammatory profile is clearly correlated with its behavior as inhibitor of intracellular calcium mobilization. The profile and potency of this compound may have relevance for the inhibition of the inflammatory response at different levels and may represent a new approach to the development of new anti inflammatory drugs. PMID- 12732368 TI - Effect of sesquiterpene lactones on the expression of the activation marker CD69 and of IL-2 in T-lymphocytes in whole blood. AB - We used flow cytometry to investigate the inhibitory effect of sesquiterpene lactones (SLs) on T-cell activation measured by the expression of its early marker CD69, and on interleukin (IL)-2, a mediator of activation, in whole blood. SLs are biologically active compounds found especially in plants from the Asteraceae family. Overnight treatment of blood with these substances led to the inhibition of CD69 and IL-2 expression. Interestingly, bifunctional SLs showed a weaker activity than monofunctional substances, which is in contradiction with the data obtained so far, using other biological test systems. Additionally, SLs did not completely inhibit CD69 or IL-2 expression. We also determined their toxicity and observed only a low effect. Up to now, studies on cytotoxicity have only been performed using cultured cell lines. From these results it may be supposed that these natural compounds preferentially show toxic effects towards transformed cell lines. Altogether, the results demonstrated that SLs effectively inhibit the activation of the T-lymphocyte response in whole blood and proved the utility of a whole blood system in studying their biological effects. PMID- 12732370 TI - 1st Joint Meeting of the International Bone and Mineral Society and the Japanese Society for Bone and Mineral Research. 3-7 June 2003, Osaka, Japan. Abstracts. PMID- 12732369 TI - Effect of acute betaine administration on hepatic metabolism of S-amino acids in rats and mice. AB - Alterations of hepatic glutathione level by betaine were observed previously. In this study effects of betaine administration (1000 mg/kg, i.p.) on S-amino acid metabolism in rats and mice were investigated. Hepatic glutathione level decreased rapidly followed by marked elevation in 24 hr. Concentrations of S adenosylmethionine, S-adenosylhomocysteine, and methionine were increased whereas cystathionine decreased significantly, suggesting that homocysteine generated in the methionine cycle is preferentially remethylated to methionine rather than being utilized for synthesis of cysteine. Hepatic cysteine concentration declined immediately, but plasma cysteine increased. Effect of betaine on hepatic cysteine uptake was estimated from the difference in cysteine concentration in major blood vessels connected to liver. Cysteine concentration either in the portal vein or abdominal aorta was not altered, however, a significant increase was noted in the hepatic vein, indicating that hepatic uptake of cysteine was decreased by betaine treatment. Activities of glutamate cysteine ligase, cystathionine beta-synthase, and cystathionine gamma-lyase were elevated in 24 hr. Pretreatment with propargylglycine, an irreversible inhibitor of cystathionine gamma-lyase, did not abolish the betaine-induced reduction of hepatic glutathione in 4 hr, however, the elevation at t=24 hr was blocked completely. In conclusion the present results indicate that betaine administration induces time-dependent changes on hepatic metabolism of S-amino acids. Betaine enhances metabolic reactions in the methionine cycle, but inhibits cystathionine synthesis and cysteine uptake, leading to a decrease in supply of cysteine for glutathione synthesis. Reduction in glutathione is subsequently reversed due to induction of cysteine synthesis and glutamate cysteine ligase activity. PMID- 12732372 TI - Cognitive therapy versus applied relaxation as treatment of generalized anxiety disorder. AB - Cognitive therapy (CT) and applied relaxation (AR) as treatments of generalized anxiety disorder (GAD) were compared in a sample of 45 patients of a community mental health center, randomly allocated to condition. Patients were assessed before and after a 12-session treatment, and at one and six months follow-ups. There was a 20% drop out from CT and 15% from AR (NS), with some drop outs being considerably improved. Both completers and intention-to-treat analyses revealed that both treatments were effective (ESs of composite and specific measures ranging from 0.53 to 1.14). At one-month follow-up AR tended to do better than CT, with CT catching up with AR at six months. Recovery rates and proportions of patients showing reliable change were comparable to other studies on AR and CT, with 55% of CT and 53.3% of AR patients recovered on the STAI-trait at six-month follow-up. These results confirm that both CT and AR are effective treatments for GAD, and also that there is still room for improvement. PMID- 12732373 TI - Anger-associated panic attacks in Cambodian refugees with PTSD; a multiple baseline examination of clinical data. AB - Despite the increasing recognition of the importance of anger as a key aspect of post-traumatic stress disorder (PTSD), the presence of anger-induced panic attacks has been understudied in traumatized groups. The present investigation determines the prevalence of anger-associated panic attacks among Cambodian refugees suffering from PTSD. Specific characteristics of these episodes that were examined included frequency, symptoms, and cognitions (in particular, fear of death from bodily dysfunction). In a survey of 100 Khmer patients suffering PTSD, 58% reported anger-associated panic attacks in the last month. These attacks occurred at a mean rate of 6.2 attacks a month and were characterized by extreme arousal and in 81% of these cases, fears of death due to bodily dysfunction during the anger-induced panic. Mechanisms for this high rate of fear of death during anger arousal are discussed with a focus on culture-specific catastrophic cognitions. PMID- 12732374 TI - The Meta-Cognitive Beliefs Questionnaire: development of a measure of obsessional beliefs. AB - A 67 item self-report questionnaire called the Meta-Cognitive Beliefs Questionnaire (MCBQ) was developed to assess endorsement of beliefs about the importance of control and negative consequences associated with unwanted, ego dystonic intrusive thoughts, images and impulses. The MCBQ and a battery of questionnaires that assessed symptoms and cognitions of worry, obsessive compulsive disorder, anxiety, and depression were administered to large samples of undergraduate students. Beliefs about control of intrusive thoughts and perceived negative consequences due to uncontrolled mental intrusions had a unique significant relationship with obsessions, and to a lesser extent, worry. These findings are consistent with current cognitive behavioral theories that suggest an important role for meta-cognitive beliefs in the pathogenesis of obsessions. PMID- 12732375 TI - Impact of case formulation in cognitive behaviour therapy for psychosis. AB - Case formulation (CF) is the 'first principle' in cognitive behavior therapy (CBT), and the cornerstone of CT for psychosis. Experiment 1 (E1) assesses impact of CF in CBT for psychosis on perception of therapeutic relationship. Experiment 2 assesses the impact of CF on strength of delusional and self-evaluative beliefs (E2). Both E1 and E2 assess impact of CF on symptoms of anxiety and depression. CF had little or no impact on these variables. Descriptive data suggested there were other benefits for certain participants. Therapists' perception of the therapeutic relationship did improve following CF. Six clients found CF, at least in part, a negative experience. PMID- 12732376 TI - "Not just right experiences": perfectionism, obsessive-compulsive features and general psychopathology. AB - Individuals with obsessive-compulsive disorder (OCD) frequently report uncomfortable sensations of things not being just right ("not just right experiences"; NJREs) and a need to ritualize until they quiet these sensations. However, very little work has been conducted to empirically examine the nature and characteristics of this phenomenon. In this paper, we present two studies that systematically examine NJREs in large undergraduate samples. In Study 1, NJREs were shown to be related to both OCD features and maladaptive domains of perfectionism. In Study 2, NJREs were again significantly related to OCD features. Further, NJREs were shown to have particularly strong relationships with specific OCD symptom clusters (e.g., checking, ordering). Finally, NJREs were significantly more strongly correlated with features of OCD than symptoms from other domains of psychopathology (e.g., social anxiety, trait anxiety, worry, depression). General conclusions are drawn, and future directions are discussed. PMID- 12732377 TI - Anger, aggression, and risky behavior: a comparison of high and low anger drivers. AB - This research tested hypotheses from state-trait anger theory applied to anger while driving. High and low anger drivers drove equally often and as many miles, but high anger drivers reported more frequent and intense anger and more aggression and risky behavior in daily driving, greater anger in frequently occurring situations, more frequent close calls and moving violations, and greater use of hostile/aggressive and less adaptive/constructive ways of expressing anger. In low impedance simulations, groups did not differ on state anger or aggression; however, high anger drivers reported greater state anger and verbal and physical aggression in high impedance simulations. High anger drivers drove at higher speeds in low impedance simulations and had shorter times and distances to collision and were twice as likely to crash in high impedance simulations. Additionally, high anger drivers were more generally angry. Hypotheses were generally supported, and few gender differences were noted for anger and aggression. PMID- 12732378 TI - Dissociative experiences are related to commissions in emotional memory. AB - Two rival hypotheses of the link between dissociative experiences and emotional memory were tested: 1) the defence mechanism hypothesis that assumes that dissociation promotes fragmentation of emotional memory; and 2) the fantasy proneness hypothesis that claims that because of its overlap with fantasy proneness, dissociation is related to commissions in emotional memory. Undergraduates scoring high (n = 19) or low (n = 19) on the Dissociative Experiences Scale (DES) listened to an aversive story. Next, a free recall task was administered. While high dissociators made more commission errors in their free recall than did low dissociators, fantasy proneness did not account for this difference. PMID- 12732379 TI - Suppressing unwanted memories: where there is a will, there is a way? AB - Research suggests that suppressing unwanted thoughts, generally, is an ineffective thought control strategy, because suppression attempts oftentimes fail, and, furthermore, result in a paradoxical increase of unwanted thoughts, later on. The present study sought to investigate whether manipulated expectations about suppression efficacy determine actual effects of suppression attempts. To test this hypothesis, participants listened to an audiotaped story, and were subsequently appointed to one of four conditions: a no-instruction control (n = 20), suppression (n = 20), suppression-works (n = 20; participants were told that suppression generally is a fruitful strategy), or suppression-does not-work (n = 25; participants were told that suppression primarily has paradoxical effects) condition. Two hours later, participants' memories of the story were tested, and several metamemory questions were answered. Induced expectations actually determined the perceived efficacy of suppression attempts, as well as thought frequency, although perceived or actual accuracy of recollections was not affected by the instructions. PMID- 12732380 TI - Further exploration memory bias in compulsive washers. AB - The aim of the present study was to replicate Radomsky and Rachman's findings on memory bias in obsessive-compulsive disorder (OCD), using the same procedure but an increased sample size, more specific control groups, and a full analysis of contamination attribution data. Sixteen OCD-washers, 16 OCD-checkers, 16 social phobic patients and 16 non-anxious controls were presented with 50 'clean' or 'dirty' objects. After this incidental encoding phase, participants were asked to freely recall the objects, to rate their anxiety when almost touching each object, and, finally, to attribute each object to one of the two contamination conditions ('clean' or 'dirty'). Verbal episodic memory was also assessed with the California Verbal Learning Test. The results indicate that, contrary to Radomsky and Rachman's findings, OCD-washers did not differ from the other participants in their memory for 'clean' and 'dirty' objects. However, the OCD washers recalled more accurately the 'dirty' source of contamination than the 'clean' source. This result was specific to the OCD-washers, and suggests a memory bias for contextually threatening information. The differences between our findings and those published by Radomsky and Rachman's are discussed. PMID- 12732381 TI - A review on the diagnosis, natural history, and treatment of familial hypercholesterolaemia. AB - BACKGROUND: Familial hypercholesterolaemia (FH) affects approximately 1 in 500 people (10 million world-wide) and the elevated serum cholesterol concentrations lead to a more than 50% risk of fatal or non-fatal coronary heart disease by age 50 years in men and at least 30% in women aged 60 years. Based on a systematic literature search, we review the natural history of FH, describe the diagnostic criteria, and consider the effectiveness of treatment. METHODS: A comprehensive review was conducted of the literature on the diagnosis of FH, the morbidity and mortality related to treated and untreated FH, and the evidence on the effectiveness of treatment of FH in adults and children. Treatment options have changed since statin treatment became available, and we have not considered pre statin therapy studies of treatment effectiveness. FINDINGS AND DISCUSSION: A clinical diagnosis of FH is widely used, but a definitive diagnosis can be made by genetic screening, although mutations are currently only detected in 30-50% of patients with a clinical diagnosis. Under-diagnosis of FH has been reported world wide ranging from less than 1% to 44%. The relative risk of death of FH patients not treated with statins is between three and fourfold but treatment is effective, and delays or prevents the onset of coronary heart disease. Early detection and treatment is important. Aggressive LDL therapy is more effective in the regression of the carotid intima media thickness than conventional LDL therapy. Diagnosis at birth is problematic, and should be delayed until at least 2 years of age. Statins are not generally recommended for the treatment of children up to adolescence. Resins may be used but poor adherence is a problem. Technical advances in mutation detection, and the identification of other genes that cause FH, are likely to have important implications for the cost effectiveness of genetic diagnosis of FH. PMID- 12732382 TI - Localization of genes that control LDL size fractions in baboons. AB - LDL phenotypes are strongly associated with risk of cardiovascular disease and are heritable, although little is known about individual genes that influence them. We investigated genetic control of LDL size-related phenotypes in 634 pedigreed baboons fed three diets contrasting in levels of fat and cholesterol. On a high-cholesterol high-fat diet, we obtained significant evidence for a quantitative trait locus (QTL) for cholesterol concentrations of lipoproteins between 27 and 28 nm (LOD=4.22, genomic P=0.0047) on the baboon homologue of human chromosome 22. For baboons fed a low-cholesterol high-fat diet, we obtained suggestive evidence for a QTL for cholesterol concentrations between 26 and 27 nm (LOD=2.67) on the baboon homologue of human chromosome 5. We speculate that this QTL influences LDL size distributions because LDL median diameters and other LDL fractions also showed peak LOD scores in this same chromosomal region. On a low cholesterol low-fat basal diet we obtained suggestive evidence for a QTL for cholesterol concentrations of lipoproteins between 26 and 27 nm in diameter (LOD=2.15) on the baboon homologue of human chromosome 16. Thus, we have evidence for three putative QTLs that influence variation in baboon LDL size phenotypes on different diets. PMID- 12732383 TI - Sarpogrelate HCl, a selective 5-HT2A antagonist, retards the progression of atherosclerosis through a novel mechanism. AB - Although sarpogrelate HCl is widely used for the prevention of arterial thrombosis, its effect on atherosclerosis is unknown. Accordingly, we here investigated the effects of sarpogrelate HCl on a rabbit model of atherosclerosis. Male rabbits were fed a 0.5% cholesterol diet (HCD) (Gp 1), HCD with vitamin E (Gp 2), HCD with vitamin E and sarpogrelate (Gp 3), or HCD with sarpogrelate alone (Gp 4) for 8 weeks. The atherosclerotic area was decreased by feeding of vitamin E and sarpogrelate (16.9+/-2.0% in Gp 1 vs. 8.2+/-2.0% in Gp 3). Tone-related basal NO release was higher in Gps 3 and 4. Acetylcholine induced relaxation tended to be improved in Gp 3. The amount of eNOS mRNA was increased in Gp 4, and aortic cyclic GMP concentration showed the same tendency. O(2)(-) release tended to be decreased in Gps 2 and 3. The matrix metalloproteinase-1 (MMP-1)-positive area was decreased, and the percentage ratio of cell numbers of smooth muscle cells/macrophages in the plaque was increased in Gp 3. The results demonstrated that sarpogrelate HCl retards the progression of atherosclerosis in rabbits, and that this effect is enhanced by concomitant administration of vitamin E. Although upregulation of eNOS may play a role as one of the underlying mechanisms, our results suggest that an additional mechanism possibly involving the antiproliferative effects of sarpogrelate HCl on smooth muscle cells and macrophages-may also play an important role. PMID- 12732384 TI - Immunity to heat shock protein 65--an additional determinant in intimal thickening. AB - Inflammation occurring consequent to vessel injury is thought to play an important role in atherosclerosis and restenosis. Autoimmunity to HSP65 has been shown to accelerate early atherogenesis in rabbits and mice, whereas in humans epidemiological data support this contention. In the current study, we explored the possibility of HSP65 influencing the extent of neointimal growth in the rat carotid injury model. Rats were either immunized with recombinant mycobacterial HSP65, heat killed preparation of Mycobacterium tuberculosis (MT), or with PBS, all emulsified in incomplete Freund's adjuvant. Animals were boosted with a similar protocol 3 weeks following the primary immunization and 2 weeks later carotid injury was applied in all animals by balloon inflation. Upon sacrifice 2 weeks later, sera were obtained for measurement of anti-HSP65 antibodies by ELISA, splenocytes were assessed for proliferative response to in vitro priming with HSP65, and carotid arteries were removed for evaluation of neointimal growth. Rats immunized with HSP65 exhibited a brisk and sustained humoral immune response to HSP65, and cellular immunity was also evident by thymidine uptake to splenocytes primed with the respective protein. Neointimal/medial ratio was significantly increased in HSP65 immunized rats, in comparison with MT injected and control animals. In conclusion, immunity to HSP65 can play a role in accelerating restenosis following arterial injury. These results should be further investigated in humans as they may provide a possible link between infections and restenosis/accelerated arteriosclerosis. PMID- 12732385 TI - Homocysteine strongly enhances metal-catalyzed LDL oxidation in the presence of cystine and cysteine. AB - Here we show that homocysteine stimulates low density lipoprotein (LDL) oxidation at copper(II) concentrations causing only a slight oxidation of LDL lipids. LDL oxidation by homocysteine and copper(II) is further enhanced in the presence of cystine, although cystine alone does not stimulate LDL oxidation with copper(II). Similarly, a combination of cysteine with homocysteine provoked a more than additive increase of oxidation. Simultaneous presence of cysteine and homocystine also resulted in a more than additive oxidative effect which was not statistically significant, however. Stimulation of LDL oxidation in the presence of homocysteine by cystine was also observed with iron(III) at acidic pH and when LDL oxidation was initiated by azo-compound generated peroxyl radicals. At pH 7.4 histidine is able to prevent LDL oxidation by copper(II) in a thiol mixture similar to the one found in human plasma if present in tenfold excess over homocysteine, but loses its inhibitory effect at higher homocysteine concentrations. The synergistic effect on metal-catalyzed LDL oxidation observed with mixtures of homocysteine and cystine or cysteine sustains the hypothesis that the epidemiological association between raised homocysteine levels and risk of cardiovascular disease is caused by an increase in oxidative stress. PMID- 12732386 TI - Aortic recruitment of blood lymphocytes is most pronounced in early stages of lesion formation in apolipoprotein-E-deficient mice. AB - Lymphocyte accumulation in the arterial intima affects development of atherosclerotic lesions. We studied the kinetics of lymphocyte accumulation in the arterial wall by injecting lymphocytes from male LacZ transgenic mice into female apolipoprotein-E-deficient mice. Recipient mouse aortas were removed and separated into lesioned and non-lesioned parts 2, 24, 48, or 72 h later. The accumulation of donor lymphocytes was quantified with real-time PCR of donor lymphocyte-specific genes. The accumulation of lymphocytes in the lesioned parts of aorta decreased with increasing lesion severity (r=-0.74, P=0.0005, n=18). Moreover, the accumulation of lymphocytes in the lesioned part of aorta was larger (392+/-108%, P=0.016) compared with the accumulation in the non-lesioned part in mice with mild atherosclerosis, whereas it was smaller (58+/-19%, P<0.01) compared with the accumulation in the non-lesioned part in mice with severe atherosclerosis. The results suggest that aortic recruitment of blood lymphocytes is most pronounced in early stages of lesion formation. PMID- 12732387 TI - The influence of psychological stress and socioeconomic status on platelet activation in men. AB - OBJECTIVE: Circulating monocyte- and neutrophil-platelet aggregates are sensitive markers of in vivo platelet activation. Socioeconomic status is inversely associated with risk of coronary heart disease. We assessed the impact of psychological stress on leukocyte-platelet aggregates in men from higher and lower socioeconomic status groups. METHODS: Participants were 37 healthy non smoking men aged 30-59 years, divided by occupation into higher and lower social status groups. Blood was drawn at baseline, immediately following stressful behavioural tasks, and at 30 and 75 min post-stress, and aggregates were analysed using flow cytometry. Cardiovascular and subjective stress responses were also monitored. RESULTS: There were significant increases following stress in monocyte , neutrophil-, lymphocyte- and total leukocyte-platelet aggregates (all P<0.05). The largest responses were in monocyte-platelet (21% increase) and neutrophil platelet (16.7% increase) aggregates. Lower socioeconomic status men had greater numbers of leukocyte-platelet aggregates throughout, but the magnitude of stress responses did not vary with social status. The increase in monocyte- and leukocyte-platelet aggregates was associated with systolic blood pressure stress responsivity. CONCLUSIONS: Psychological stress induces platelet activation as indexed by leukocyte-platelet aggregates, and correlations with cardiovascular stress reactions suggest that sympathoadrenal responses may be responsible. Platelet activation may be a mechanism through which social position influences cardiovascular disease risk. PMID- 12732388 TI - Vitamin C, diclophenac, and L-arginine protect endothelium-dependent vasodilation against elevated circulating fatty acid levels in humans. AB - An acute elevation of circulating non-esterified fatty acids (NEFAs) has previously been shown to impair endothelium-dependent vasodilation (EDV). In this study, we investigated if local administration of vitamin C (n=8, 18 mg/min), L arginine (n=8, 12.5 mg/min), or the cyclooxygenase (COX) inhibitor diclophenac (n=8, 0.5 mg/min) can counteract the endothelial dysfunction seen during infusion of Intralipid plus heparin (n=10). EDV and endothelium-independent vasodilation (EIDV) were studied in the forearm after local administration of methacholine chloride (Mch; 2 and 4 microg/min) and sodium nitroprusside (SNP; 5 and 10 microg/min). Forearm blood flow (FBF) was determined with venous occlusion plethysmography. Intralipid and heparin increased circulating NEFA levels sevenfold and impaired EDV (P<0.001 vs baseline). Concomitant administration of L arginine or diclophenac abolished the NEFA-induced impairment in EDV. Concomitant vitamin C administration actually improved EDV (P<0.05 vs baseline). NEFA elevation increased EIDV (P<0.01), but this effect was not significant after L arginine or diclophenac infusions. In conclusion, an acute elevation of circulating NEFAs led to impaired EDV. Administration of L-arginine, vitamin C or COX inhibition abolished this effect, suggesting that NEFAs might interact with endothelial vasodilatory function through multiple mechanisms. PMID- 12732389 TI - Increased expression of T cell activation markers (CD25, CD26, CD40L and CD69) in atherectomy specimens of patients with unstable angina and acute myocardial infarction. AB - Atherosclerotic plaques contain a chronic immune mediated inflammation in which T cells play an important role. A previous study revealed that the numbers of interleukin-2 receptor-positive T cells is increased in culprit lesions of patients with acute coronary syndromes; a finding of considerable interest since it indicates a recent change in the intraplaque T cell mediated immune response. Confirmation of this observation is important, because it could provide insight into the onset of the acute event. We have, therefore, expanded our earlier work by using a panel of different T cell activation markers (CD25, CD26, CD40L, CD69). The study is based on 58 culprit lesions from patients who underwent coronary atherectomy. There were four groups of patients: chronic stable angina (n=13), stabilized unstable angina (n=16), refractory unstable angina (n=15), and acute myocardial infarction (AMI; n=14). Activated T cells were expressed as a percentage of the total of CD3-positive cells. CD25, CD26, CD40L, and CD69/CD3 percentages increased with the severity of the coronary syndrome. In patients with AMI all percentages were significantly higher than in patients with chronic stable angina. CD25, CD26, CD40L, and CD69/CD3 percentages in patients with an unstable condition (refractory unstable angina and AMI) were significantly higher than those in patients with a stable condition (chronic stable or stabilized unstable angina) The finding that the percentage of T cells with recent onset activation is significantly increased in the culprit lesions of patients with acute coronary syndromes suggests strongly that a recent change in pathogenic stimulation has occurred leading to local T cell activation. PMID- 12732390 TI - Apolipoprotein C-III, a strong discriminant of coronary risk in men and a determinant of the metabolic syndrome in both genders. AB - AIMS: Apolipoprotein C-III (apoC-III) has been recognized as a useful marker of triglyceride-rich lipoproteins (TRLs) metabolism and proposed as an indicator of prognosis for coronary risk in healthy subjects. We studied cross-sectionally in a population having low cholesterol levels, but a high prevalence of the metabolic syndrome, whether serum levels of total apoC-III or its sub-fractions were independent markers of prevalent coronary heart disease (CHD) or were related to variables reflecting the metabolic syndrome. METHODS AND RESULTS: In 857 unselected participants of the representative population sample of the Turkish Adult Risk Factor Survey in 2001, apoC-III as well as other risk variables were evaluated, and CHD was diagnosed based on clinical findings and Minnesota coding of resting electrocardiograms. The sample consisted of men and women, aged 33-82 years, having a mean waist circumference of 89.4 and 92.9 cm, respectively, 42% of whom had the metabolic syndrome identified by criteria of the ATP III. ApoC-III values were measured by turbidimetric immunoassay. Mean concentrations for non-high-density lipoprotein (nonHDL) apoC-III in men and women were 6.4 and 6.2 mg/dl, respectively, and for apoC-III in HDL were 6.2 and 6.3 mg/dl, respectively. NonHDL apoC-III was similar to, and apoC-III in HDL was higher than that in Western populations. Both fractions of apoC-III were significantly correlated with lipids, lipoproteins, apoB, anthropometric measures, and blood pressures in both genders. Correlations for both were high with serum triglycerides (r(s)=around 0.70) and apoB (r(p)=around 0.37). Total apoC-III as well as both fractions were significantly correlated in women also with levels of inflammatory risk markers: strongly (r=0.40-0.45, P<0.001) with complement C3, and weakly (r(s)=around 0.20, P<0.001) with C-reactive protein. A cutoff of >7.0 mg/dl as opposed to lower levels of nonHDL apoC-III indicated the presence of hypertriglyceridemic hyperapo B with an age-adjusted odds ratio (OR) of 13.8; it indicated the presence of metabolic syndrome with 4.66-fold likelihood. Total apoC-III and nonHDL apoC-III proved to be significantly (P trend <0.05 and 0.002) and strongly associated with prevalent CHD in men even after adjustment for age, low-density lipoprotein-cholesterol (LDL-C) and high density lipoprotein-cholesterol (HDL-C): OR gradients across upper and lower quartiles were 3.88-fold (CI: 1.3; 11.4) and 8.8-fold (CI: 2.6; 29.8), respectively. CONCLUSIONS: In a population among whom the metabolic syndrome prevails, total- and nonHDL apoC-III are each a determinant in both genders of the metabolic syndrome and of hypertriglyceridemic hyperapo B. Each is a powerful significant marker of prevalent CHD in men independent of LDL- and HDL-C levels. In women, despite being correlated with inflammatory risk markers, the significant association of elevated levels of apoC-III with CHD did not prove to be independent of age. PMID- 12732391 TI - Does elevated body mass modify the influence of postmenopausal estrogen replacement on atherosclerosis progression: results from the estrogen in the prevention of atherosclerosis trial. AB - OBJECTIVE: To determine whether the estrogen-related reduction in atherosclerosis progression demonstrated in the estrogen in the prevention of atherosclerosis trial (EPAT) is modified by body mass index (BMI). STUDY DESIGN: Subgroup analyses were performed using data from EPAT, a randomized, double-blind, placebo controlled trial designed to determine whether unopposed 17beta-estradiol administered for a 2-year treatment period reduces the progression of subclinical atherosclerosis in healthy postmenopausal women. The primary trial endpoint was the rate of change of common carotid artery intima-media thickness (IMT). In this subgroup analysis, the sample was divided into 122 women with BMI<30 kg/m(2) and 77 women with BMI> or =30 kg/m(2). Statistical analysis was performed using mixed general linear models to evaluate whether the treatment effects on IMT progression rates differed in the two BMI groups. RESULTS: There was no significant difference in the estradiol treatment effect on IMT progression rates between postmenopausal women with BMI<30 vs. > or =30 kg/m(2) (P=0.52). In the 77 subjects who did not use lipid-lowering therapy, there was significant improvement in IMT with estradiol treatment that was evident in both BMI groups (P=0.48 for differences between BMI groups). CONCLUSIONS: In contrast to the epidemiological observation that obese postmenopausal women do not derive benefit from estrogen replacement therapy, results of this study indicate that estradiol treatment is beneficial in preventing progression of atherosclerosis regardless of initial BMI. CONDENSATION: Estradiol treatment is beneficial in preventing progression of atherosclerosis in postmenopausal women not receiving lipid lowering therapy, regardless of their initial body mass index. PMID- 12732392 TI - High oxidative stress in patients with stable coronary heart disease. AB - Oxidized low density lipoprotein (oxLDL) plays a pivotal role in the development of atherosclerosis. The aim of the study was to investigate the relationship between oxLDL and other oxidative stress biomarkers with stable coronary heart disease (CHD). We compared the degree of oxidative stress in patients with CHD and sex-matched healthy control subjects in a case-control study. The study included 64 male subjects: 32 patients with stable CHD and 32 normal control subjects. Levels of circulating oxLDL were measured by a monoclonal antibody 4E6 based competition ELISA. Comparison of oxidative stress marker levels between cases and controls, adjusted for age, revealed significantly higher plasma oxLDL levels (63.32+/-25.49 vs. 37.73+/-20.58 U/l, P=0.001), lower serum levels of autoantibodies against oxLDL (341.53+/-350.46 vs. 796.45+/-1034.2 mU/ml, P=0.021), higher activities of the antioxidant enzymes superoxide dismutase in erythrocytes (951+/-70.2 vs. 771.6+/-191.2 U/g, P=0.032) and glutathione peroxidase in whole blood (GSH-Px: 10714.4+/-3705.4 vs. 5512.2+/-1498.1 U/l, P<0.001). The risk of having CHD was 20.6-fold greater (95% CI, 1.86-228.44, P=0.014) in the highest tertile of the oxLDL distribution than in the lowest, determined by logistic regression analysis on the combined study population after adjustment for age and other potential confounding factors. When the risk associated with GSH-Px levels was calculated, the odds ratio was 305.3 (95% CI, 5.07-18369.95, P=0.006) in the highest tertile compared with the lowest. Our results showed that an oxidative stress occurs in patients with CHD despite being clinically stable and under medical treatment. The combination of oxLDL levels and GSH-Px activity may be useful for the identification of patients with stable CHD. PMID- 12732393 TI - Decrease in LDL size in HIV-positive adults before and after lopinavir/ritonavir containing regimen: an index of atherogenicity? AB - BACKGROUND: Hypertriglyceridemia (HTG) is frequently observed during highly active antiretroviral therapy (HAART) including protease inhibitor. Apolipoprotein (apo) CIII could be involved in this HTG by inhibition of triglyceride (TG) hydrolysis, which leads to the occurrence of small dense low density lipoprotein (sdLDL), a recognized cardiovascular risk factor. OBJECTIVE: To characterize the influence of lopinavir/ritonavir-containing regimen on lipoprotein profile. DESIGN AND METHODS: 24 antiretroviral-experienced HIV infected adults (including 14 patients in therapeutic interruption of at least 2 months) and 14 HIV uninfected healthy controls were enrolled. Serum lipid parameters (total cholesterol (TC), HDL-C, LDL-C, TG, apoA1, apoB, apoCIII), lipoprotein composition and LDL size were determined before initiation of lopinavir/ritonavir-containing regimen, and at 1 and 3 months thereafter. RESULTS: At baseline an atherogenic lipid profile was evidenced, characterized by a moderate HTG associated to a smaller mean LDL size (25.16 vs 25.93 nm, P<0.001), an enrichment in TG of LDL (11.4 vs 6.0%, P<0.01) and a high prevalence of sdLDL (75 vs 7%, P<0.01) when compared to controls. After 1 month of lopinavir/ritonavir-containing regimen, a significant reduction of LDL size (24.81 vs 25.16 nm, P<0.05) and a significant increase in cholesterol total (5.53 vs 4.49 mmol/l, P<0.001), in TG (4.20 vs 2.01 mmol/l, P<0.001), in apoA1 (1.28 vs 1.11 g/l, P<0.001), in apoB (1.08 vs 0.94 g/l, P<0.01), in apoCIII (0.16 vs 0.10 g/l, P<0.001), in TG percentage in LDL (14.4 vs 11.4, P<0.05) and in TG percentage in HDL (10.2 vs 8.3, P<0.05) were observed. CONCLUSIONS: Advanced stage of HIV infection is associated with an atherogenic lipid profile including a high prevalence of sdLDL. Lopinavir/ritonavir-containing regimen accentuates the reduction of LDL size. Since fibrates decrease TG and increase LDL size, they appear as a logical option to manage HAART-induced HTG. PMID- 12732394 TI - Influence of exonic polymorphisms in the gene for LDL receptor-related protein (LRP) on risk of coronary artery disease. AB - The low density lipoprotein (LDL) receptor-related protein (LRP) is a multifunctional receptor involved in numerous biological processes relevant to vascular biology including lipoprotein metabolism. Several polymorphisms in the LRP gene have been described and in this study we examined their influence on coronary artery disease (CAD). We compared the frequencies of the exon 3 (C766T), exon 6 (C663T), exon 22 (C200T), and four rarer and more recently described polymorphisms in approximately 600 Caucasian subjects aged <50 years with angiographic CAD and approximately 700 similarly aged subjects without symptomatic CAD randomly selected from the community. We found the distribution of exon 22 C200T genotypes to differ significantly between the CAD (CC: 52%, CT: 39%, TT: 9%) and control subjects (CC: 43%, CT: 46%, TT: 11%, P=0.005), with the CC genotype conferring an odds ratio (OR) for CAD of 1.5 (95% CI: 1.2-1.8, P=0.001) despite a lack of significant influence on plasma cholesterol or triglyceride. The other LRP polymorphisms were less common. Two showed an association with CAD; for the exon 3 C766T polymorphism the TT genotype was significantly lower (1.0 vs. 2.7%; OR: 0.36; P=0.04) and, for the exon 6 C663T polymorphism, the heterozygote frequency was higher (6.2 vs. 3.4%; OR: 1.9; P=0.03) in CAD subjects. In conclusion, LRP gene polymorphisms, particularly the relatively common exon 22 C200T polymorphism, are a significant risk factor for premature CAD in Caucasians. PMID- 12732395 TI - Double blind, randomized study of estradiol replacement therapy on markers of inflammation, coagulation and fibrinolysis. AB - Estrogen replacement therapy (ERT) has been found to be associated with increased cardiovascular risk in the first year after initiation of ERT. We compared the effects of oral and transdermal estradiol (E2) replacement therapy on markers of inflammation, coagulation and fibrinolysis in a randomized double-blind trial. Forty-three healthy women were randomized 6 weeks after surgically induced menopause to receive treatment with either oral or transdermal E2 over a period of 28 weeks. At baseline and after 28 weeks, levels of serum lipids and lipoproteins, and markers of coagulation, fibrinolysis and inflammation were determined. Among fibrinolytic parameters, oral E2 shortened euglobulin clot lysis time (P<0.05) and reduced tissue type plasminogen activator antigen (P=0.01) and plasminogen activator inhibitor activity (P<0.05). Among coagulation parameters, both routes of E2 replacement decreased fibrinogen levels (P=0.002 for oral and P=0.007 for transdermal E2). Oral E2 resulted in an increase in C reactive protein (CRP) from 2.15 (0.71-4.05) to 3.41 (1.12-5.92) mg/l (P=0.04), while transdermal E2 showed no effect. Levels of serum amyloid A (SAA), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) did not change significantly after oral and transdermal E2. Oral E2 significantly improved the lipid profile, while transdermal E2 had a less pronounced effect. Both oral and transdermal E2 significantly reduced fasting glucose. Oral E2 was associated with a pro-inflammatory response, but at the same time improved fibrinolytic capacity, showed no pro-coagulatory effects, and acted beneficially on lipids and lipoproteins. There was no influence of transdermal E2 on markers of coagulation activation, fibrinolysis and inflammation, but it decreased fibrinogen levels significantly. Further studies are needed to explore the clinical relevance of these observations. PMID- 12732396 TI - A novel functional polymorphism in the PECAM-1 gene (53G>A) is associated with progression of atherosclerosis in the LOCAT and REGRESS studies. AB - A 53G>A polymorphism identified in the 5' untranslated region (5'UTR) of the platelet endothelial cell adhesion molecule-1 (PECAM-1) gene alters a putative shear stress responsive element (SSRE). PECAM-1 was shown to be responsive to shear stress and transient transfection of human umbilical vein endothelial cell (HUVECs) with two luciferase reporter constructs driven by the PECAM-1 promoter and 5'UTR showed a response of the 53G allele, not the 53A allele, to shear stress. Association between the 53G>A, and the previously published L125V polymorphism, and coronary atherosclerosis was examined in two angiographic studies. The frequencies of the rare alleles of the 53G>A and L125V polymorphisms were 0.01 and 0.49, respectively, in the Lopid Coronary Angiography Trial (LOCAT) study and 0.02 and 0.49, respectively, in the Regression Growth Evaluation Statin Study (REGRESS) study. Compared with 53G homozygotes, carriers of the 53A allele showed less focal progression of disease in the LOCAT study and a similar trend in the diffuse progression of disease in the REGRESS study, whereas no association between L125V and coronary atherosclerosis was observed in either study. These data demonstrate that the PECAM-1 gene is responsive to shear stress in vitro and that decreased PECAM-1 gene expression in 53A carriers may influence reduced progression of vessel stenosis in patients with coronary artery disease. PMID- 12732398 TI - The signal sequence polymorphism of the MnSOD gene is associated with the degree of carotid atherosclerosis. AB - Redox-state of the cells of vascular walls is an important determinant of atherosclerosis. Manganese superoxide dismutase (MnSOD) is an essential anti oxidant enzyme working in mitochondria of mammalian cells. A potentially functional amino acid polymorphism (Ala16Val) has been described in the signal sequence of the enzyme. The aim of the current study was to test whether the signal sequence polymorphism of the MnSOD would be associated with the degree of carotid atherosclerosis. The polymorphism was genotyped in a sample of 989 middle aged hypertensive and control subjects. Carotid atherosclerosis was quantified as intima-media thickness (IMT) by ultrasound. The signal sequence polymorphism was found to be a minor determinant of carotid IMT explaining 1.3% of the overall variation, the Val allele associated with the higher IMT. In women, a significant interaction with plasma levels of low-density lipoprotein (LDL) cholesterol was detected, since LDL cholesterol levels were positively correlated with carotid IMT only in the carriers of the Val allele and the Val allele was associated with higher IMT only in the subjects with highest plasma levels of LDL cholesterol. In conclusion, the signal sequence polymorphism of the MnSOD gene is a minor determinant of carotid IMT pointing out the importance of redox-balance in the atherogenesis. PMID- 12732397 TI - Oral and transdermal estrogens both lower plasma total homocysteine in male-to female transsexuals. AB - Plasma total homocysteine (tHcy) levels are on average lower in women versus men, indicating an estrogenic effect. Oral estrogens (absorbed via the liver) may be hypothesized to have stronger effects on hepatic homocysteine metabolism than transdermal estrogens. We randomly assigned 30 male-to-female transsexuals (20-44 years old) to 4 months' administration of oral ethinyl estradiol (n=15) or transdermal 17beta-estradiol (n=15), both with the antiandrogen cyproterone acetate (CA). Ten other male controls were treated with CA only. At baseline and after 2 and 4 months, plasma tHcy was analyzed in conjunction with plasma folate. Oral ethinyl estradiol and transdermal 17beta-estradiol similarly reduced plasma tHcy (geometric mean 10.6 micromol/l [95% CI 8.2-13.9] to 7.5 [6.5; 8.8], and 11.3 [8.1; 16.4] to 8.4 [6.5; 11.1]; P<0.001 for both), whereas CA had no effects. No effects were found on folate levels. Thus, oral and transdermal estrogens decrease plasma tHcy to a similar degree (by geometric mean -26%), which suggests that a hepatic mechanism is unlikely to play an important role in the decline of tHcy levels. PMID- 12732399 TI - Detection of Chlamydia pneumoniae but not of Helicobacter pylori in symptomatic atherosclerotic carotids associated with enhanced serum antibodies, inflammation and apoptosis rate. AB - BACKGROUND AND PURPOSE: Numerous seroepidemiological and pathological studies linked Chlamydia pneumoniae and Helicobacter pylori with atherosclerosis. However, analyses of these infectious agents in the pathogenesis of stroke are either lacking or contradictory. Therefore, we evaluated the detection rate of C. pneumoniae and H. pylori in normal carotids vs. atherosclerotic carotids and compared these findings with serology, plaque morphology, inflammatory cell infiltrates and apoptosis rate. METHODS: The study was performed on 40 morphological normal carotids from autopsy and 20 advanced atherosclerotic carotids from endarterectomy after stroke. Serum IgG antibody titre was measured by enzyme immunoassay (H. pylori) and microimmunofluorescence (MIF) technique (C. pneumoniae). Immunohistochemistry (IHC) and Western blotting were performed to identify C. pneumoniae, H. pylori, to characterize plaque morphology (macrophages and smooth muscle cells) and the inflammatory infiltrate (T- and B cells) and to detect apoptosis (TUNEL staining). RESULTS: C. pneumoniae was found significantly more frequently in atherosclerotic than in normal carotids (P=0.001), which correlated with elevated C. pneumoniae IgG-antibody titres (P=0.048). Although H. pylori was not detected in carotids, elevated H. pylori antibody titres were significantly associated with the degree of atherosclerosis (P=0.001). The C. pneumoniae infected carotids displayed a slightly enhanced infiltrate of T cells and apoptosis rate, but no morphological changes. CONCLUSION: C. pneumoniae but not H. pylori, was detected by IHC primarily in symptomatic carotids, without specific morphological differences. Correlation of C. pneumoniae in-situ detection and IgG antibodies suggested a possible connection between respiratory tract and endovascular infection. The C. pneumoniae associated T-lymphocytes and apoptosis rate indicate an immune-mediated inflammatory process, involving vascular walls. PMID- 12732400 TI - Serum uric acid in hypertensive patients with and without peripheral arterial disease. AB - BACKGROUND: Uric acid is frequently elevated in hypertension. In addition to renal and metabolic disturbances, lower limb ischemia might contribute to hyperuricemia among hypertensives complicated by peripheral arterial disease (PAD). OBJECTIVE: To test the hypothesis that uric acid status is related to lower limb function in hypertensives with PAD. METHODS: Serum and 24-h urine uric acid levels and other risk factors were examined in 145 hypertensives free of PAD and 166 hypertensives with PAD. Ankle/brachial index (ABI) and absolute claudication distance (in PAD) on a treadmill test (ACD) were assessed. RESULTS: In multiple regression analysis for serum uric acid in the total group, PAD emerged as an independent determinant (P=0.03) next to age (P=0.005), triglycerides (P=0.04), and insulin (P=0.02). Serum uric acid concentrations were higher in hypertensives with PAD (404+/-101 vs. 347+/-80 micromol/l, P<0.001) independent of components of the metabolic syndrome (body mass index, triglycerides, insulin) and of age, gender, diabetes mellitus, pulse pressure, cholesterol, C-reactive protein, and treatment. After adjustment for kidney function by uric acid/creatinine ratio, values remained higher in hypertensives with PAD (P=0.01). Uric acid excretion was higher in the PAD group (P<0.001), whereas uric acid clearance was comparable between both groups. In multiple regression analysis for ACD (357+/-183 m) in the PAD group, serum uric acid (P=0.02), C-reactive protein (P<0.0001), age (P=0.02), and smoking (P=0.004) were independently associated. ABI (0.62+/-0.17) was not related to uric acid in PAD patients. CONCLUSION: Hyperuricemia is more pronounced in hypertensives complicated by PAD and is associated with worse functional status of the peripheral circulation. PMID- 12732402 TI - Troponin I concentrations following primary percutaneous coronary intervention predict large infarct size and left ventricular dysfunction in patients with ST segment elevation acute myocardial infarction. AB - The aim of this study was to investigate the ability of troponin I (cTnI) levels to predict myocardial infarction size in patients with ST-segment elevation acute myocardial infarction (STEMI) undergoing primary percutaneous coronary intervention (PCI). In 87 patients with STEMI undergoing primary PCI, serial plasma concentrations of cTnI and alpha-hydroxybutyrate deshydrogenase (HBDH) were measured before PCI and over the following 72 h. Enzymatic infarct size was estimated by the cumulative release of HBDH during the 72 h following PCI (QHBDH72). Delayed radionuclide left ventricular ejection fraction (LVEF) was measured in 63 patients. While cTnI concentrations at admission did not correlate with QHBDH72 or with LVEF, from the 3rd to the 72nd h following PCI, they did correlated with QHBDH72 (P<0.001; R: 0.76-0.86) and with LVEF (P<0.001; R: -0.42 to -0.50). Receiver-operator characteristic (ROC) curve analysis showed that admission concentrations of cTnI could not predict either a large infarct size (i.e., QHBDH72>10 g-eq l(-1)) or a low LVEF (i.e., LVEF<40%). However, 6 h and up until 72 h after PTCA, cTnI concentrations were predictive of large enzymatic infarct size (sensitivity: 91 and 95%, specificity: 90 and 87%, respectively) and of LVEF under 40% (sensitivity: 75 and 77%, specificity: 90 and 78%, respectively). Thus, our study suggests that in contrast with admission cTnI concentration, cTnI levels following primary PCI represent a reliable tool for predicting large enzymatic infarct size and may help in selecting patients with a high risk of low LVEF at 1 month. PMID- 12732401 TI - Combined effect of coenzyme Q10 and fenofibrate on forearm microcirculatory function in type 2 diabetes. AB - Arteriopathy is the principal complication of type 2 diabetes mellitus. It develops from endothelial dysfunction, which we have hypothesised occurs in diabetes primarily as a consequence of dyslipidaemia and oxidative stress. Fenofibrate and CoQ may improve endothelial function by regulating dyslipidaemia and oxidative stress, respectively. We therefore aimed to assess the independent and combined effects of fenofibrate and coenzyme Q(10) (CoQ) on endothelium dependent and endothelium-independent vasodilator function of the forearm microcirculation in type 2 diabetes. Eighty dyslipidaemic type 2 diabetics were randomized to receive fenofibrate (200 mg/daily), CoQ (200 mg/daily), fenofibrate plus CoQ (200+200 mg daily), or placebo for 12 weeks. Forearm microcirculatory function was assessed with venous occlusion plethysmography during the infusion of acetylcholine (ACh), bradykinin (BK), sodium nitroprusside (SNP) and N(G) monomethyl-L-arginine (L-NMMA) into the brachial artery. Blood flow responses were calculated as area under the curve (AUC). Fenofibrate significantly lowered plasma cholesterol, triglyceride and fibrinogen (P<0.001), and elevated HDL cholesterol and homocysteine (P<0.001). CoQ did not change plasma isoprostanes, but significantly lowered systolic blood pressure and HbA(1c) (P<0.05). Fenofibrate plus CoQ significantly improved (P<0.05) the AUC for ACh, BK and SNP without significantly altering basal responses to L-NMMA. Fenofibrate or CoQ alone did not significantly alter blood flow responses. Improvements in blood flow were independent of changes in plasma lipids, blood pressure, homocysteine and isoprostanes, but were correlated (P=0.013) with HbA(1c). In conclusion, in this factorial trial we found that only the combination of fenofibrate and CoQ markedly improved endothelial and non-endothelial forearm vasodilator function in dyslipidemic type 2 diabetic patients. The favourable vascular effect of this therapeutic combination could be due to increase in the bioactivity of and/or responses to endothelium-derived relaxing factors, including nitric oxide, and this may entail synergistic stimulation of peroxisome proliferator-activated receptors. PMID- 12732404 TI - Transporting efficacious treatments from academia to community: barriers and opportunities. PMID- 12732405 TI - Single versus multiple drug focus in substance abuse clinical trials research: the devil is in the details. PMID- 12732403 TI - Single versus multiple drug focus in substance abuse clinical trials research. AB - Complex patterns of multiple substance use pose clinical and methodological challenges for substance abuse clinical trials research. To increase measurement precision and internal validity, the modal approach has been to target both treatment interventions and outcome assessment to a single class of abused substance. This strategy warrants reconsideration because it entails limitations in recruitment feasibility and generalization of study findings. This report reviews pros and cons of single versus multiple targeted drugs, suggests guidelines for choosing between these strategies and outlines methods for broadening the scope of substance abuse clinical trails to take abuse of multiple substances into account. We recommend that investigators consider moving away from a single drug focus in three ways. First, include systematic assessment of a wide range of psychoactive substance use throughout the trial and evaluate the impact of study treatments on use of all classes of drugs. Second, except where contraindicated, include patients who use and abuse multiple classes of substances even in trials evaluating treatment of a single targeted drug. Third, consider inclusion of polysubstance abusers or those who primarily abuse multiple classes of substances in the same clinical trial. Although many treatment efficacy questions can best be answered by single focus studies, we recommend that such designs be adopted only after less restrictive designs are first considered. PMID- 12732406 TI - Can we design and replicate clinical trials with a multiple drug focus. PMID- 12732407 TI - Income distribution and risk of fatal drug overdose in New York City neighborhoods. AB - Accidental drug overdose is a substantial cause of mortality for drug users. Neighborhood-level factors, such as income distribution, may be important determinants of overdose death independent of individual-level factors. We used data from the Office of the Chief Medical Examiner to identify all cases of accidental deaths in New York City (NYC) in 1996 and individual-level covariates. We used 1990 US Census data to calculate the neighborhood-level income distribution. This multi-level case-control study included 725 accidental overdose deaths (cases) and 453 accidental deaths due to other causes (controls) in 59 neighborhoods in NYC. Overdose deaths were more likely in neighborhoods with higher levels of drug use and with more unequal income distribution. In multi-level models, income maldistribution was significantly associated with risk of overdose independent of individual-level variables (age, race, and sex) and neighborhood-level variables (income, drug use, and racial composition). The odds of death due to drug overdose were 1.63-1.88 in neighborhoods in the least equitable decile compared with neighborhoods in the most equitable decile. Disinvestment in social and economic resources in unequal neighborhoods may explain this association. Public health interventions related to overdose risk should pay particular attention to highly unequal neighborhoods. PMID- 12732408 TI - The effects of escalating doses of smoked cocaine in humans. AB - This study examined the effects of escalating doses of smoked cocaine in non treatment-seeking cocaine users. Cocaine sessions were conducted twice per day for 3 consecutive days. During each session, one group of participants smoked a 12 mg cocaine dose, followed by a 25 mg dose, followed by four 50 mg doses at 14 min intervals (escalating-dose group); another group of participants smoked six 50 mg cocaine doses at 14 min intervals (fixed-dosing group). Cocaine produced dose-dependent increases in heart rate (HR), blood pressure, ratings of positive drug effect, e.g., "good drug effect", and ratings of cocaine dose, e.g., "liking", in the escalating-dose group. The 50 mg dose also increased these measures in the fixed-dosing group to a level that was not different than that observed following the initial 50 mg dose in the escalating-dose group. The largest effects of 50 mg cocaine were observed following the initial dose, with the latter 50 mg doses maintaining but not increasing these effects in both groups. The effects of cocaine in both groups were consistent for most measures within a day and between days. Resting baseline heart rate, blood pressure and cocaine craving were lower during the first session and higher thereafter. These results demonstrate that increasing the dose of cocaine during a bout, i.e., "binge", of cocaine use can increment the effects of cocaine, but administering the same cocaine dose maintains, but does not increment the effects of cocaine. PMID- 12732409 TI - Injection frequency mediates health service use among persons with a history of drug injection. AB - Drug injectors are known to have high rates of hospital and emergency room (ER) use. We hypothesized that out-of-treatment injection drug users (IDUs) have higher rates of health service use than methadone-maintained persons, and that heroin injection frequency mediates health service use among drug injectors (IDUs). HIV-negative individuals with a history of drug injection were recruited from a needle exchange program (NEP) and a methadone maintenance treatment program (MMT) in Providence, RI. ER visits, outpatient visits, and hospitalizations in the last 6 months were the dependent variables with number of heroin injections the hypothesized mediator variable. The 472 participants were predominately male (60.6%) and white (82%) with a mean age of 37. NEP clients were more likely than MMT clients to visit an ER (39.2% vs. 29.8%; P=0.03). NEP participants were somewhat more likely to report a hospital admission (16.0% vs. 10.6%; P=0.08). Relative to non-injecting MMT participants, the odds of visiting an ER were 1.80 and 1.67 times higher for subjects recruited through NEP and actively injecting MMT participants, respectively. Additionally, subjects recruited through NEP (OR=2.2) and actively injecting MMT participants (OR=2.3) were over twice as likely to report a hospital admission than non-injecting MMT participants. Each increase of one heroin injection per day increased the expected odds of injection-related infection by a factor of 1.92. NEP clients are more likely to have ER visits and hospitalizations than methadone clients. We describe a pathway by which injection frequency influences health service use. PMID- 12732410 TI - A tripartite of HIV-risk for African American women: the intersection of drug use, violence, and depression. AB - The intersection of drug use, violence, and depression with HIV-risk among African American women is an under explored area of research. The current analyses examine whether particular sexual risk behaviors are associated with exposure to violence, depression or both among 420 African American out-of treatment female drug users. Women were stratified into four mutually exclusive groups: drug users with exposure to violence (n=64), drug users with clinical depression (n=62), drug users with both (n=41), and drug users only (n=253). Multinomial logistic regression analyses examined the association of demographics and sexual risk behaviors across the tripartite groups. Women with a history of sexually transmitted diseases were more likely to experience violence and depression both alone and jointly. Women who had two or more sexual partners in the last 30 days (OR=2.26) and women who had an early onset of alcohol use (OR=2.50) were at an increased risk for having the full tripartite of drug use, violence and depression. Never being married was a protective factor for the full tripartite. As expected, more risk factors were found among women who had the full tripartite than among women with one or two of the factors. The co-existence of the tripartite factors and sexual risk behaviors may indicate a need to ultimately provide more specialized prevention and intervention efforts to combat HIV infection. This area of research may improve our understanding of the numerous obstacles to HIV intervention among drug-using populations. PMID- 12732412 TI - Smoking and substance abuse in outpatients with schizophrenia: a 2-year follow-up study in Turkey. AB - BACKGROUND: This study aimed to determine the prevalence of smoking and substance abuse in outpatients with schizophrenia, and to determine the relationship between smoking status and sociodemographic/clinical characteristics of schizophrenia. METHODS: One hundred and thirty-six schizophrenic outpatients were assessed by the structured clinical interview for DSM-IV (SCID) and brief psychiatric rating scale (BPRS). Demographic and treatment variables were obtained from case records and interviews with patients and their family members. RESULTS: The frequency of smoking among schizophrenic patients was 50%. However, the rate of substance abuse was 5.2%. We found no statistically significant differences between the smoker and the non-smoker patients on the demographic variables of age, sex, marital status, and employment status. There was no significant difference between the groups on BPRS scores. However, smokers were receiving a higher daily dose of neuroleptic than non-smokers. CONCLUSION: The prevalence of smoking among schizophrenic patients was similar to the rates found in Western cultures. However, the prevalence of substance abuse was lower in Turkish patients as compared with patients in the Western population. PMID- 12732411 TI - Screening for cannabis use disorders in an adolescent emergency department sample. AB - Cannabis, more often than alcohol, is the drug mentioned in substance-related reasons for treatment of an adolescent in an emergency department (ED). This study examined the prevalence of DSM-IV cannabis and alcohol diagnoses in an adolescent ED sample, evaluated the performance (i.e. sensitivity and specificity) of DSM-IV cannabis symptoms and other screening items as indicators of cannabis diagnosis status, and examined parent-adolescent agreement on the presence of cannabis and alcohol diagnoses. Adolescents (ages 13-19, n=442) admitted to an ED for a non-substance-related injury were administered the diagnostic interview schedule for children (DISC). Parents (n=272) of adolescents younger than age 18 completed the DISC-parent version to report on their child's drug use. A minority met criteria for a current DSM-IV cannabis or alcohol diagnosis: 7.9% for both alcohol and cannabis, 7.5% for cannabis-only, and 9.0% for alcohol-only. Frequency of cannabis use had the best overall performance in discriminating those with and without a cannabis diagnosis compared with items on perceived risk of cannabis use, peer cannabis use, and alcohol and cigarette use. Parent reports generally underestimated the adolescent's substance use. Questions on level of substance use generally provide an efficient method of screening adolescents for substance-related problems in an ED setting. PMID- 12732414 TI - Alcohol outcome expectancies and risk for alcohol use problems in women with and without a family history of alcoholism. AB - Studies have shown that alcohol expectancies are positively associated with drinking and alcohol abuse and dependence symptoms among alcohol users. This study looked at the relationship of alcohol expectancies, family history of alcoholism, menstrual cycle and drinking behavior. The present study compared alcohol expectancies using the Alcohol Expectancy Questionnaire (AEQ) in 85 women ranging from 18 to 35 years of age. Forty-one women had a confirmed parental history of alcoholism (family history positive, FHP) and 44 women had no parental history of alcoholism (family history negative, FHN). Participants' mood, alcohol consumption, and daily consequences of alcohol use were prospectively tracked across one menstrual cycle. Alcohol expectancies at screening were significantly greater in FHP women in four of the six AEQ subscales, as well as the composite score. Alcohol expectancies correlated significantly with drinking behavior among FHN women. In FHP women, alcohol expectancies were elevated regardless of their drinking level. Alcohol expectancies decreased among FHP women, but not FHN women, after prospectively tracking their drinking behavior and consequences of drinking. Negative outcomes of drinking were increased among the FHP women who were heavy drinkers. Irrespective of family history status, alcohol use in moderate drinkers increased significantly during menses compared to the follicular and luteal phases of the menstrual cycle. These findings suggest that menstrual cycle may also play a role in alcohol consumption. Thus, the results of the present study indicate that issues related to the level of alcohol consumption, menstrual cycle phase and family history of alcoholism should be considered when addressing alcohol abuse in women. PMID- 12732413 TI - Survival study of opioid addicts in relation to its adherence to methadone maintenance treatment. AB - OBJECTIVES: To study the putative role of methadone maintenance treatment in the improvement of life expectancy of opioid addicts. DESIGN: Retrospective longitudinal study. PARTICIPANTS: All 1487 patients receiving methadone maintenance treatment in Alicante between June 1990 and December 1997. STATISTICAL ANALYSIS: Mortality rates were studied using Kaplan-Meier survival curves. Protection or risk factors were analyzed using Cox's proportional hazards model. RESULTS: Mortality rates decreased from 87/1000 in 1991 to 17/1000 in 1997. The following factors influenced mortality: HIV infection [Hazard Ratio (HR)=7, 95% confidence interval (CI)=4-12]; current methadone status (HR=3.2, 95%CI=1.5-7.1) and MMT retention (retained vs. drop-out, HR=0.5, 95%CI=0.2-1.1; re-enrolled vs. drop-out, HR=0.3, 95%CI=0.2-0.5). CONCLUSION: Expediting entry and re-enrolling in methadone maintenance treatment improves survival. PMID- 12732415 TI - Preliminary study on reducing oral moist snuff use. AB - BACKGROUND: Tobacco exposure reduction may be an alternative treatment approach for those tobacco users who are unwilling or unable to quit tobacco use. However, very little information is available on the feasibility of this type of intervention, especially in the area of oral moist snuff tobacco (ST). This pilot study examined whether reducing ST use using various methods can be achieved and whether this reduction results in lower exposure to carcinogens. METHODS: Moist snuff users (N=40 males) were randomly assigned to 4 mg nicotine gum, non-tobacco mint snuff, brand switching, or elimination of ST use in specific situations. These approaches were used to reduce ST use or nicotine exposure by at least 25% for the first 2 weeks and 50% the subsequent 6 weeks of treatment. Follow-up sessions occurred at 12 and 26 weeks. RESULTS: Significant reductions were observed in tins per week and cotinine levels across all conditions. Among the intent-to-treat population, the abstinence rate was 15% at 26 weeks. Reduction in nicotine exposure was associated with reduction in exposure to nitrosamines. CONCLUSION: Reduction in ST use may be a viable approach for those oral moist ST users with no immediate quit plans. Future research in this area is needed. PMID- 12732416 TI - Low-shear modeled microgravity: a global environmental regulatory signal affecting bacterial gene expression, physiology, and pathogenesis. AB - Bacteria inhabit an impressive variety of ecological niches and must adapt constantly to changing environmental conditions. While numerous environmental signals have been examined for their effect on bacteria, the effects of mechanical forces such as shear stress and gravity have only been investigated to a limited extent. However, several important studies have demonstrated a key role for the environmental signals of low shear and/or microgravity in the regulation of bacterial gene expression, physiology, and pathogenesis [Chem. Rec. 1 (2001) 333; Appl. Microbiol. Biotechnol. 54 (2000) 33; Appl. Environ. Microbiol. 63 (1997) 4090; J. Ind. Microbiol. 18 (1997) 22; Curr. Microbiol. 34(4) (1997) 199; Appl. Microbiol. Biotechnol. 56(3-4) (2001) 384; Infect Immun. 68(6) (2000) 3147; Cell 109(7) (2002) 913; Appl. Environ. Microbiol. 68(11) (2002) 5408; Proc. Natl. Acad. Sci. U. S. A. 99(21) (2002) 13807]. The response of bacteria to these environmental signals, which are similar to those encountered during prokaryotic life cycles, may provide insight into bacterial adaptations to physiologically relevant conditions. This review focuses on the current and potential future research trends aimed at understanding the effect of the mechanical forces of low shear and microgravity analogues on different bacterial parameters. In addition, this review also discusses the use of microgravity technology to generate physiologically relevant human tissue models for research in bacterial pathogenesis. PMID- 12732417 TI - Evaluation of QIAamp DNA Stool Mini Kit for ecological studies of gut microbiota. AB - Cell lysis efficiency and the quality of DNA extracts from complex bacterial ecosystems are two major concerns in molecular ecological studies of gut microbiota. In this study, we use PCR-denaturing gradient gel electrophoresis (DGGE) DNA profiling, random cloning and sequence analysis of 16S rRNA genes to compare the QIAamp DNA Stool Mini Kit with the bead beating technique in the preparation of DNA extracts from gut microbiota of pigs. We also developed a washing procedure that can release more than 93% of bacterial cells attached to the gut mucosa. Both the QIAamp kit and bead beating method lysed approximately 95% of bacterial cells. PCR-DGGE DNA profiles of ileal and cecal microbiota from both digesta and mucosa that were generated from the DNA extracts using the two methods were nearly identical. Random cloning and sequence analysis also demonstrated the high quality of DNA extracts using the two methods. Two random clone sets of 16S rRNA genes generated from the DNA extracts had a similar degree of bacterial diversity. Different preparations of DNA extract from a single sample using the QIAamp kit consistently produced similar PCR-DGGE DNA profiles with similarity indexes higher than 99%. Our data suggest the appropriateness of the QIAamp DNA Stool Mini Kit for the studies of gut microbial ecology and the effectiveness of the QIAamp kit in processing multiple samples for cell lysis and DNA extraction. PMID- 12732418 TI - Improved methods of isolation and purification of myxobacteria and development of fruiting body formation of two strains. AB - By using baiting techniques and different purification methods, a high number of myxobacterial strains have been isolated as pure cultures from soil of different regions of China. Because myxobacterial cells do not disperse easily in liquid media, a medium containing an enzymatic hydrolysate of casein (CEH) medium have been used for purification and purity tests combined in a single step. The key method, in which isolates are reintroduced to sterile rabbit dung to induce fruiting bodies formation, facilitates purification of myxobacteria. Sterile rabbit dung pellets are used to mimic the natural growth substance of these organisms which has the advantage that characteristic fruiting bodies emerge, which is a key characteristics in the taxonomy of myxobacteria. In this study, the optimum program of isolation and purification of some myxobacteria strains has been established which will facilitate screening programs. Moreover, the development of fruiting body formation of strain BD20 (Chondromyces) and strain BD54 (Cystobacter) have been recorded in this study. PMID- 12732419 TI - An immunomagnetic separation-real-time PCR method for quantification of Cryptosporidium parvum in water samples. AB - The protozoan parasite Cryptosporidium parvum is known to occur widely in both raw and drinking water and is the cause of waterborne outbreaks of gastroenteritis throughout the world. The routinely used method for the detection of Cryptosporidium oocysts in water is based on an immunofluorescence assay (IFA). It is both time-consuming and nonspecific for the human pathogenic species C. parvum. We have developed a TaqMan polymerase chain reaction (PCR) test that accurately quantifies C. parvum oocysts in treated and untreated water samples. The protocol consisted of the following successive steps: Envirochek capsule filtration, immunomagnetic separation (IMS), thermal lysis followed by DNA purification using Nanosep centrifugal devices and, finally, real-time PCR using fluorescent TaqMan technology. Quantification was accomplished by comparing the fluorescence signals obtained from test samples with those from standard dilutions of C. parvum oocysts. This IMS-real-time PCR assay permits rapid and reliable quantification over six orders of magnitude, with a detection limit of five oocysts for purified oocyst solutions and eight oocysts for spiked water samples. Replicate samples of spiked tap water and Seine River water samples (with approximately 78 and 775 oocysts) were tested. C. parvum oocyst recoveries, which ranged from 47.4% to 99% and from 39.1% to 68.3%, respectively, were significantly higher and less variable than those reported using the traditional US Environmental Protection Agency (USEPA) method 1622. This new molecular method offers a rapid, sensitive and specific alternative for C. parvum oocyst quantification in water. PMID- 12732420 TI - A quantitative analysis of DNA extraction and purification from compost. AB - We quantified both DNA and humic acid concentrations during the extraction and purification of DNA from compost. The DNA extraction method consisted of bead beating with SDS for cell lysis, poly(ethylene glycol)-8000 precipitation for preliminary DNA purification, and chromatography on a 10-ml Sephadex G-200 column for final DNA purification. Direct microscopic observation of pre- and post-lysis samples revealed that 95.3+/-2.3% of native cells was lysed. Sixty-three percent of the original DNA was lost during purification, resulting in a final DNA yield of 18.2+/-3.8 microg DNA/g of wet compost. The humic acid content was reduced by 97% during the purification steps resulting in a final humic acid concentration of 27+/-4.7 ng humic acid/microl. The purified DNA fragments were up to 14 kbp in size and were sufficiently free of contaminants to allow both restriction enzyme digestion by four different enzymes and PCR amplification of 16S rDNA. PMID- 12732421 TI - Immunodetection of pneumolysin in human urine by ELISA. AB - An ELISA test has been employed for the detection of pneumolysin (PLY) in urine from 14 pneumococcal pneumonia patients and from 11 healthy adult volunteers. The urines of all the 11 healthy adult volunteers developed signals around the mean of the blanks, whereas all the pneumococcal pneumonia patient urines rendered signals at least five times this mean. Chemiluminescent Western blot analyses of these urines, carried out with the PLY-specific rabbit polyclonal IgG preparation used in ELISA, were negative. The 30-kDa filtrates of three high-signal urines were ELISA negative, suggesting that this ELISA test mainly detected high molecular weight forms in urine rather than free PLY-derived antigenic fragments. The urine sample, which rendered the highest ELISA signal, was then concentrated by filtration through a 10-kDa filter. When this concentrate was subjected to Western blot with the ELISA-capture monoclonal antibody, a major band was developed. Its relative molecular mass was similar to that of recombinant PLY and its peptide mass fingerprinting showed peptides corresponding to amino acid stretches from the four domains of the PLY molecule. When the pool of PLY negative urines was sham-contaminated with purified recombinant pneumolysin, a conspicuous matrix effect was observed; nevertheless, this ELISA test was still reproducible and highly sensitive, detecting pneumolysin in the order of picograms per milliliter. A comparison was also made between this PLY-ELISA and the Binax NOW Streptococcus pneumoniae Urinary Antigen Test in analysing bacterial isolates. On the basis of the minimum number of pneumococci examined, both tests were shown to have similar potency, but strain-dependent discrepancies were observed. This ELISA could provide an alternative to the Binax NOW Streptococcus pneumoniae Urinary Antigen Test in the diagnosis of pneumococcal pneumonia. PMID- 12732422 TI - A comparison of DNA profiling techniques for monitoring nutrient impact on microbial community composition during bioremediation of petroleum-contaminated soils. AB - Amplicon length heterogeneity PCR (LH-PCR) and terminal restriction fragment length polymorphisms (TRFLP) were used to monitor the impact that nutrient amendments had on microbial community dynamics and structural diversity during bioremediation of petroleum-contaminated soils. Slurried soils contaminated with petroleum hydrocarbons were treated in airlift bench-scale bioreactors and were either amended with optimal inorganic nutrients or left unamended. Direct DNA extraction and PCR amplification of whole eubacterial community DNA were performed with universal primers that bracketed the first two or three hypervariable regions of the 16S rDNA gene sequences. The LH-PCR method profiled a more diverse microbial community than did the TRFLP method. The LH-PCR method also tracked differences between the communities due to nutrient amendments. An in silico database search for bacterial genera with amplicon lengths represented in the community fingerprints was performed. It was possible to qualitatively identify different groups in the microbial community based on the amplicon length variations. A similar "virtual" search was performed for the TRFLP fragments using the web-based TAP-TRFLP program. Cloning and sequencing of the PCR products confirmed the in silico database matches. The application of the LH-PCR method as a monitoring tool for bioremediation could greatly enhance and extend the current understanding of the microbial community dynamics during the biodegradation of environmental contaminants. PMID- 12732423 TI - Monitoring promoter activity in a single bacterial cell by using green and red fluorescent proteins. AB - We investigated the possibility of monitoring promoter activity with flow cytometry by using green fluorescent protein (GFPmut2) and red fluorescent protein (drFP583) in a single bacterial cell. The drFP583 was used as an intrinsic marker of the bacterial cells, because it was expressed constantly in Escherichia coli MC1061 strain. The GFPmut2 expressed under the control of the Hg(2+) ion inducible mer promoter/operator, was used to study promoter activity. Over 75% of the cells were positive for red and green fluorescence in flow cytometric analysis. The average green fluorescence of the whole population increased from 6.7 to 1700 when the mercury concentration was increased from 0 to 1 x 10(-4) M, while the red fluorescence was unaffected by the mercury concentration. These results show that gfpmut2 and drFP583 could be expressed under different promoters in one bacterial cell and measured independently with a flow cytometer. PMID- 12732424 TI - Identification and expression of human CD81 gene on murine NIH/3T3 cell membrane. AB - The human CD81 (hCD81) molecule has been identified as a putative receptor for hepatitis C virus (HCV). In this study, eukaryotic expression vector pCDM8-hCD81 containing hCD81 cDNA and pSV2neo helper plasmid was used to cotransfect with lipofectamine into murine fibroblast cell line NIH/3T3 to establish an hCD81 expressing cell line. Resistant cell clones were obtained 20 days after the selection with neomycin (600 micro/ml) and then cultured as monoclones. The expression of the transfected hCD81 gene in the cells was verified by RT-PCR and flow cytometry analyses. One of the selected cell clones showed obvious expression of hCD81 and was named NIH/3T3-hCD81. Competitive inhibition tests indicated that the binding of monoclonal anti-hCD81 (JS-81) to NIH/3T3-hCD81 cells was inhibited by recombinant HCV E2 protein, suggesting that the expressed hCD81 molecules on NIH/3T3-hCD81 cells maintain natural conformation of binding to HCV E2. The transfected NIH/3T3-hCD81 cells should be of great potential value in studies on HCV attachment and onset of infection. PMID- 12732425 TI - Identification of EPEC and non-EPEC serotypes in the EPEC O serogroups by PCR RFLP analysis of the fliC gene. AB - Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the flagellin gene (fliC) was performed in 233 strains of enteropathogenic Escherichia coli (EPEC) O serogroups for determining their flagellar antigen (H) status. The serological detection of flagellin is the basis for the H-codes typing system in E. coli. Thus, it is impossible to serotype nonmotile bacteria (i.e. to assign H-codes). Twenty-eight fliC restriction patterns were obtained for motile (H2, H4, H6, H7, H8, H9, H10, H11, H12, H18, H21, H27, H32, H34, H35, H40 and H51) and nonmotile serotypes (H(-)). Each motile serotype was characterized by one or two fliC specific restriction patterns. The only exception was serogroup O128ab, where a common restriction pattern was found for serotypes O128ab:H2 and O128ab:H35, even after digestion with RsaI, AluI and Sau3AI endonucleases. These two serotypes were, however, discriminated by single strand conformation polymorphism (SSCP) analysis of RsaI restriction fragments. Nonmotile strains showed fliC restriction patterns identical to some known H serotypes. The PCR-RFLP analysis of fliC gene proved to be a useful method for identifying the H variants in motile and nonmotile EPEC O serogroups. PMID- 12732426 TI - A rapid method for identifying diversity within PCR amplicons using a heteroduplex mobility assay and synthetic polynucleotides: application to characterisation of dsRNA elements associated with Cryptosporidium. AB - A 173-bp fragment of the small extra-chromosomal double-stranded RNA (dsRNA) element of Cryptosporidium parvum was generated by reverse transcriptase PCR from nucleic acid extracted from whole faeces of 18 epidemiologically unrelated cases of cryptosporidiosis. Eleven different sequences were detected and two selected as reference DNA in a heteroduplex mobility assay (HMA). Although sequence diversity was detected, this was difficult to characterise because of the similarity in electrophoretic mobility of the homo- and heteroduplex bands. A PCR method was devised to generate synthetic polynucleotides of greater sequence diversity for use in the HMA. The presence of the synthetic 173-bp fragments was enriched by using, as template for the PCR, material excised from the area of the heteroduplex bands in stained electrophoresis gels. Nine novel sequences were generated and evaluated as reference sequences in the HMA. One of these with 20 bp different from the original sequence was selected for use in the HMA for improved resolution of heteroduplex and homoduplex bands and number of patterns easily resolved (nine different patterns corresponding to different DNA sequences). This method may be useful for analysis of DNA where there is limited natural variation or little sequence variation is described. PMID- 12732427 TI - Comparative studies on the activity of basil--an essential oil from Ocimum basilicum L.--against multidrug resistant clinical isolates of the genera Staphylococcus, Enterococcus and Pseudomonas by using different test methods. AB - The essential oil basil is obtained from the aerial parts of Ocimum basilicum L. After gas chromatographic separation, the following components were identified: linalol (54.95%), methylchavikol (11.98%), methylcinnamat (7.24%) and linolen (0.14%). The activity of basil against multidrug resistant clinical isolates from the genera Staphylococcus, Enterococcus and Pseudomonas has been studied. For this purpose, standard and modified broth macrodilution methods were used and time kill kinetic of basil was studied. The minimum inhibitory concentrations (MICs) were reported between 0.0030% and 0.0007% (v/v). These concentrations were compared with the inhibitory concentrations (ICs) and the logs of the bacterial counts reduction both obtained by basil diluted in 1% Tween (Tw) 80, saline test solution (STS) and spiritus vini (Sv) 95 degrees instead in a broth. The data, obtained after application of different methods of investigation and validated with membrane filtration, showed a strong inhibitory effect of basil on the test bacteria. The chosen bacteria are widespread and pose serious therapeutic difficulties because of their high level of resistance. For this reason, the results obtained were considered encouraging. PMID- 12732429 TI - Improved fast gas chromatography for FAME analysis of bacteria. AB - Bacteria are frequently identified by fatty acid analysis. We previously reported on methods to speed up sample preparation and gas chromatography, resulting in greatly improved speed and throughput [J. Microbiol. Methods 51 (2002) 209]. In this paper, we demonstrate that further reductions in chromatographic retention times are readily achieved, leading to faster identification of bacteria. PMID- 12732430 TI - Variation between observed and true Terminal Restriction Fragment length is dependent on true TRF length and purine content. AB - Terminal Restriction Fragment (TRF) pattern analysis has become a widely used and informative tool for studying microbial communities. Variation between sequence determined or true TRF length and observed TRF length (TRF drift) has been previously reported and can significantly affect identification of bacterial species using TRF lengths predicted from sequence databases. In this study TRF drift was determined for 21 bacterial species using an ABI 310 Genetic Analyzer. TRF drift was positively correlated with true TRF length and negatively correlated with TRF purine content. This implies that subtle differences in molecular weight, whether from purine content or dye label, can significantly affect the observed TRF length. PMID- 12732428 TI - Potassium and tetraphenylphosphonium ion-selective electrodes for monitoring changes in the permeability of bacterial outer and cytoplasmic membranes. AB - A tetraphenylphosphonium ion (TPP(+))-selective electrode, originally developed as a membrane potential indicator, is useful for measuring increases in the permeability of bacterial outer membranes induced by antimicrobial agents. The combination of this electrode with a potassium ion-selective electrode enabled us to determine changes in the permeability of bacterial outer and cytoplasmic membranes simultaneously. Outer membrane permeabilization induced by antimicrobial agents, chlorhexidine and polyhexamethylene biguanide (PHMB), as monitored with the TPP(+) electrode, correlated closely with the ability of the agents to release lipopolysaccharide (LPS) from the outer membrane. PMID- 12732432 TI - A rapid and specific method to screen environmental microorganisms for cephalosporin acylase activity. AB - Medically useful semisynthetic cephalosporin antibiotics are made from precursor 7-aminocephalosporanic acid (7-ACA). Cephalosporin acylase (CA), which catalyzes hydrolysis of both glutaryl-7-aminocephalosporanic acid (GL-7ACA) and cephalosporin C (CPC) to 7-ACA, is thus a very important enzyme for producing semisynthetic beta-lactam antibiotics. To facilitate the attempts of obtaining the microorganisms with higher CA activity from natural environments, a new and specific method for screening environmental microorganisms with cephalosporin acylase activity was developed. The core part of cephalosporin was replaced by 6 amino penicillinic acid (6-APA) to generate new substrates glutaryl-6-APA and adipoyl-6-APA for screening. Serratia marcescens that is sensitive to 6-APA and resistant to penicillin G, glutaryl-6-APA and adipoyl-6-APA was used as an indicator strain in an overlaid-agar screening system. A strain capable of producing cephalosporin acylase was selected from thousands of candidates by this method. Because of its specificity, simplicity and sensitivity, the method could be easily installed into a high-throughout system. PMID- 12732431 TI - Application of NucliSens Basic Kit for the detection of Mycoplasma pneumoniae in respiratory specimens. AB - A commercially available nucleic acid sequence-based amplification (NASBA) NucliSens Basic Kit (NBK) assay for the detection of Mycoplasma pneumoniae 16S rRNA in respiratory specimens was developed and compared to standard NASBA and PCR assays previously developed in our laboratory. The specificity and sensitivity of the NBK assay was comparable to the specificity and sensitivity of the corresponding standard NASBA assay. The NBK offers standardized reagents for the development of a NASBA assay for the detection of M. pneumoniae in respiratory specimens and is easily adaptable to other amplification targets. PMID- 12732433 TI - Reliable determination of transposon insertion site in prokaryotes by direct sequencing. AB - We developed a method to identify the insertion sites of transposons in the chromosome of Salmonella using one step only. In this method, the Salmonella's genomic DNA is directly sequenced using a transposon internal primer. Reliable direct sequencing was achieved using high purity genomic DNA and an improved protocol for automated sequence machine. This note is intended to promote the use of direct sequencing, which we found to be reliable, efficient and inexpensive as compared to the other currently used methods. PMID- 12732434 TI - Special issue: microtrabecular concept of the cytoplasm revisited. PMID- 12732435 TI - The solid state cell. PMID- 12732436 TI - Joining the trek with Keith up the Serpentine Road--the lattice from another perspective. PMID- 12732437 TI - Whatever happened to the 'microtrabecular concept'? AB - Keith Porter culminated his stellar career as the founding father of biological electron microscopy by acquiring, in the late 1970s, a high-voltage electron microscope (HVEM). With this magnificent instrument he examined whole-mounts of cultured cells, and perceived within them a structured cytoplasmic matrix he named the "microtrabecular lattice". Over the next decade Porter published a series of studies, together with a team of outstanding young colleagues, which elaborated his broader "microtrabecular concept." This concept posited that microtrabeculae were real physical entities that represented the fundamental organization the cytoplasm, and that they were the physical basis of cytoplasmic motility and of cell-shape determination. The present review presents Porter's original images of microtrabeculae, after conversion to a more interpretable "digital-anaglyph" form, and discusses the rise and fall of the microtrabecular concept. Further, it explains how the HVEM images of microtrabeculae finally came to be considered as an artifact of the preparative methods Porter used to prepare whole cells for HVEM. Still, Keith's "microtrabecular concept" foretold of our current appreciation of the complexity and pervasiveness of the cytoskeleton, which has now been found by more modern methods of EM to actually be the fundamental organizing principle of the cytoplasmic matrix. During the impending eclipse of Porter's microtrabecular concept in the late 1980s, many of Keith's colleagues fondly described the cell as being filled, not with protoplasm, but with "Porterplasm." Despite the fact that Keith's view was clouded by the methods of his time, it would be fitting and apt to retain this name, still today, for the ordered matrix of cytoskeletal macromolecules that exists in the living cell. In the end, the story of what happened to Porter's microtrabecular concept should be an object lesson in scientific hubris that should humble and inform all of us in cell biology, even today--particularly when we begin to think that our most recent methods and observations are achieving "the last word". PMID- 12732438 TI - Women's health and Medicaid reform. PMID- 12732439 TI - Women's health care in the VA system: another "patchwork quilt". PMID- 12732440 TI - Availability of comprehensive women's health care through Department of Veterans Affairs Medical Center. AB - Despite increased numbers of women veterans, little is known about health services delivery to women across the Department of Veterans Affairs (VA). To assess VA availability of women's health services, we surveyed the senior clinician at each VA site serving 400 or more women veterans. We found that virtually all sites have developed arrangements, either directly or through off site contracts, to ensure availability of comprehensive women's health care. On site care, however, is routinely available only for basic services. Future work should evaluate cost and quality trade-offs between using non-VA sites to increase specialized service availability and using VA sites to enhance continuity of care. PMID- 12732441 TI - The organization and delivery of women's health care in Department of Veterans Affairs Medical Center. AB - Congressional eligibility reforms have profoundly changed the array of services to be made available to women veterans in Department of Veterans Affairs (VA) health care facilities. These include access not only to primary and specialty care services already afforded VA users, but also to a full spectrum of gender specific services, including prenatal, obstetric, and infertility services never before provided in VA settings. The implications of this legislative mandate for delivering care to women veterans are poorly understood, as little or no information has been available about how care for women veterans is organized. This article reports on the first national assessment of variations in the organization of care for women veterans. PMID- 12732442 TI - The influence of previous breast cancer upon mammography utilization. AB - Women with a previous history of breast cancer are at increased risk for developing cancer in the opposite breast. However, the literature is inconsistent regarding whether a previous history of breast cancer is associated positively with mammography utilization. Some studies indicate that women with a previous history of breast cancer are less likely to utilize mammography, although behavioral models of health care theorize that women with a history of breast cancer may be more vigilant regarding the disease. We analyzed responses from 830 women > or =50 years who participated in the 1998 National Health Interview Survey. A significantly greater proportion of women with breast cancer reported had a mammogram in the previous year (73.13%) as compared with women who did not have breast cancer (56.69%). Although a previous history of breast cancer was found to be associated positively with mammography use, women with public sources of health insurance are less likely to report mammography use. Results indicate that women with a previous history of breast cancer appear aware of the necessity for continued screening. However, enabling factors such as type of health insurance continue to exert an influence upon the utilization of mammography. PMID- 12732443 TI - Domestic violence incidents with children witnesses: findings from Rhode Island surveillance data. AB - In this study we analyze factors associated with children witnessing police reported domestic violence (DV) and determine the age distribution of children witnessing. Rhode Island Department of Health surveillance data (1996-1998) from police forms were used to assess demographic characteristics of victims, characteristics of incidents, whether children were present, and children's ages. Victim gender, age, race/ethnicity, relationship to suspect, and whether the victim was assaulted were all strong predictors of children witnessing a DV incident. Almost half (48%) of the children who witnessed DV incidents were less than 6 years old. To reach these young children, prevention and intervention programs will need to target parents and caretakers of young children and/or pediatricians. PMID- 12732444 TI - Mind control of menopause. AB - The primary objective of this study was to observe the effect of hypnosis on hot flashes (HF) and overall quality of life in symptomatic patients. A secondary objective was to observe the effect of hypnosis on fatigue. Ten healthy volunteers and four breast cancer patients (total 14 patients) with symptoms of HF were treated with four, 1 h/wk sessions of hypnosis. The same physician, with the help of a nurse, conducted every session. All subjects recorded frequency, duration, and severity of HF in a HF diary. The QLQ-C30 and Brief Fatigue Inventory forms were used to assess the impact on quality of life and fatigue, respectively. The statistical evaluations were performed, including analysis of variance and nonparametric procedures. The frequency (p < 0.0001), duration (p < 0.0001), and severity (p < 0.0001) of HF were significantly reduced. The overall quality of life was also improved (p = 0.05). The subjects enjoyed better sleep and had less insomnia (p = 0.012). There was a significant improvement on current fatigue level (p = 0.017), but we did not find a statistically significant reduction in the total fatigue level. We conclude that hypnosis appears to be a feasible and promising intervention for HF, with a potential to improve quality of life and insomnia. Although improvement in current level of fatigue was observed in this pilot study, total fatigue improvement did not reach statistical significance. PMID- 12732446 TI - Pathophysiology of human genetic CD36 deficiency. AB - CD36, originally identified as glycoprotein IV on platelets, is an 88-kDa integral membrane protein that has multiple ligands and is expressed in the cardiovascular system (ie, blood vessel walls and the heart). Human genetic CD36 deficiency is relatively frequent in Asian and African populations. Investigation into the pathophysiology of this disorder has shown that CD36 may play an important role as a major scavenger receptor for oxidized low-density lipoproteins and as a crucial transporter for long-chain fatty acids. The CD36 deficiency may be related to the phenotypic expression of the "metabolic syndrome," which is frequently associated with atherosclerotic cardiovascular diseases. It also has been reported that CD36 deficiency might be linked with cardiomyopathy. These data raised the possibility that CD36 deficiency might be an important genetic background for these life-threatening human cardiovascular diseases. PMID- 12732445 TI - Overexpression of 12-lipoxygenase and cardiac fibroblast hypertrophy. AB - Leukocyte-type 12-lipoxygenase (12-LO) catalyzes the conversion of arachidonic acid (C20:4) to 12-hydroperoxyeicosatetraenoic acid, which in turn reduces to 12 hydroxyeicosatetraenoic acid (12-HETE) by glutathione peroxidase. Results of studies in vascular smooth muscle and in adrenal glomerulosa cells have supported the concept that 12-LO is an important mediator of angiotensin II (Ang II) action. Studies also indicate that 12-HETE is a potent growth-promoting factor and facilitates proliferation in Chinese hamster ovary (CHO) fibroblast cells overexpressing the Ang II AT1a receptor (CHO-AT1a cells). However, until recently, the role of 12-LO in cardiac cells had not been explored. Cardiac fibroblasts are a major source of matrix proteins, which can lead directly to extracellular matrix deposition and cardiac fibrosis. To elucidate the role of the 12-LO pathway in fibroblast cell growth, 12-LO cDNA was stably transfected into fetal rat cardiac fibroblasts. The cells overexpressing 12-LO showed an increase in cell protein content and enlargement in cell size with a slowing of cell division rate. Furthermore, the cells overexpressing 12-LO showed increases in fibronectin and collagen deposition compared with mock-transfected cells. These features are most consistent with cellular hypertrophy instead of proliferation. It is proposed that cardiac fibroblast cells overexpressing 12-LO retain the characteristics of fibroblasts, but with additional features of myocytes that have the function of showing cell hypertrophy. These results provide the basis for proposing the hypothesis that enhanced 12-LO expression or activity could play a role in pathogenic cardiac enlargement. PMID- 12732447 TI - Molecular mechanisms of in-stent restenosis and approach to therapy with eluting stents. AB - Restenosis is the principal drawback of percutaneous coronary procedures. Until now, the only widely accepted way to reduce restenosis rate has been the stent. However, clinical restenosis still represents the major limitation of this technology. This article summarizes recent laboratory and clinical investigations concerning the mechanisms responsible for the transmission of mitogenic signals from plasma membrane to the nucleus in vascular smooth muscle cells that determine neointima formation after stent deployment. Recent experimental data on the impact of diabetes and physical exercise on restenosis also is reviewed. Finally, the new concept of local drugs that elute directly to the site of vascular injury from coated stents and the available clinical results obtained with rapamycin or paclitaxel-eluting stents are discussed. PMID- 12732448 TI - A missense mutation in the CASQ2 gene is associated with autosomal-recessive catecholamine-induced polymorphic ventricular tachycardia. AB - Catecholamine-induced polymorphic ventricular tachycardia (CPVT), a rare disease that occurs in subjects without obvious organic heart disease, is characterized by episodes of syncope, seizures, or sudden death in response to physiologic or emotional stress. This report reviews evidence that a missense mutation in the CASQ2 gene is associated with autosomal-recessive CPVT. PMID- 12732451 TI - Current and future applications of SAGE to cardiovascular medicine. AB - The recently sequenced mammalian genomes represent unprecedented resources for advancing our understanding of human diseases. Characterizing gene expression is an important step in translating genomic sequences into clinically useful information. Currently, gene expression studies are revolutionizing the approaches taken to address both basic science and clinical questions. Two major methods have emerged for the global examination of the transcriptome: microarrays and Serial Analysis of Gene Expression (SAGE). The SAGE technique comprehensively maps gene transcription by using the genomic database, yet it remains relatively underutilized for studying cardiovascular biology. This review describes current cardiovascular studies using the SAGE technique and outlines some potential strategies for employing this powerful tool to further our understanding of the cardiovascular system in health and in disease. PMID- 12732450 TI - The role of Pitx2 during cardiac development. Linking left-right signaling and congenital heart diseases. AB - Pitx2 is a bicoid-related homeodomain transcription factor that plays a critical role in directing cardiac asymmetric morphogenesis. Ectopic Pitx2c expression in the developing myocardium correlates with double outlet right ventricle (DORV) in laterality mutants. Pitx2 loss of function experiments cause severe cardiovascular defects, such as atrial isomerism (AI), double inlet left ventricle, transposition of the great arteries (TGA), persistent truncus arteriosus (PTA), and abnormal aortic arch (AAA) remodeling. Current studies suggest that Pitx2-mediated signaling during cardiogenesis is conducted within three different cell types: the myocardium, the cardiac neural crest (CNC) cells, and the pharyngeal arch mesenchyme. Impaired Pitx2 function in discrete myocardial regions seems to lead to DORV, AI, and possibly TGA. On the other hand, impaired Pitx2 expression in the CNC leads preferentially to PTA. AAA remodeling is likely to occur owing to impaired cross-talk of the CNC cells with the pharyngeal arch mesenchyme. Thus, Pitx2 appears to be directing left-right identity to the cardiac venous components (e.g., the atria), whereas it appears to be modeling the morphologic arrangement of distinct myocardial components in the arterial pole. These data suggest that altered left-right signaling underlies the etiology of several common congenital cardiac malformations. PMID- 12732449 TI - Regulation of sarco(endo)plasmic reticulum Ca2+ adenosine triphosphatase by phospholamban and sarcolipin: implication for cardiac hypertrophy and failure. AB - The cardiac isoform of sarco(endo)plasmic reticulum Ca(2)(+) adenosine triphosphatase (SERCA2a) plays an important role in the contraction and relaxation of cardiac muscle. Phospholamban (PLN) and its homologue sarcolipin (SLN) are the endogenous regulators of SERCA2a. Evidence is accumulating that SERCA2a is intimately involved in the pathogenesis of cardiac hypertrophy and failure. Recent studies using genetically engineered animals revealed the implication of PLN for the development of cardiomyopathic phenotypes. This review focuses on advances in the understanding of molecular regulation of SERCA2a by PLN and SLN, and their implications for cardiac hypertrophy and failure in vivo. PMID- 12732452 TI - Toxicological aspects of Kampo medicines in clinical use. AB - Among 210 medicinal prescriptions used in present-day Japan, the clinical uses and the acute, chronic and mutagenic toxicity study of 16 Kampo (Japanese herbal) medicines are summarized. These Kampo medicines are classified into two categories; eight prescriptions containing Bupleurum root (Bupleurum falcatum L.) such as Sho-saiko-to and Saiko-keishi-to, and eight prescriptions not containing Bupleurum root such as Juzen-taiho-to and Ninjin-yoei-to. Studies of some potential interaction between herbal medicine and western drugs are also described. PMID- 12732454 TI - Beneficial effect of combined administration of some naturally occurring antioxidants (vitamins) and thiol chelators in the treatment of chronic lead intoxication. AB - Ameliorative effects of few naturally occurring antioxidants like ascorbic acid (vitamin C), alpha-tocopherol (vitamin E) either alone or in combination with meso-2,3-dimercaptosuccinic acid (DMSA) or monoisoamyl DMSA (MiADMSA), on parameters indicative of oxidative stress in the liver, kidney, brain and blood of lead-exposed rats were studied. Male Wistar rats were exposed to 0.1% lead acetate in drinking water for 3 months and treated thereafter with DMSA or its analogue MiADMSA (50 mg/kg, intraperitoneally), either individually or in combination with vitamin E (5 mg/kg, intramuscularly) or vitamin C (25 mg/kg, orally) once daily for 5 days. The effects of these treatments in influencing the lead-induced alterations in haem synthesis pathway, hepatic, renal and brain oxidative stress and lead concentration from the soft tissues were investigated. Exposure to lead produced a significant inhibition of delta-aminolevulinic acid dehydratase (ALAD) activity from 8.44+/-0.26 in control animals to 1.76+/-0.32 in lead control, reduction in glutathione (GSH) from 3.56+/-0.14 to 2.57+/-0.25 and an increase in zinc protoporphyrin level from 62.0+/-3.9 to 170+/-10.7 in blood, suggesting altered haem synthesis pathway. Both the thiol chelators and the two vitamins were able to increase blood ALAD activity towards normal, however, GSH level responded favorably only to the two thiol chelators. The most prominent effect on blood ALAD activity was, however, observed when MiADMSA was co administered with vitamin C (7.51+/-0.17). Lead exposure produced a significant depletion of hepatic GSH from 4.59+/-0.78 in control animals to 2.27+/-0.47 in lead controls and catalase activity from 100+/-3.4 to 22.1+/-0.25, while oxidized glutathione (GSSG; 0.34+/-0.05 to 2.05+/-0.25), thiobarbituric acid reactive substance (TBARS; 1.70+/-0.45 to 5.22+/-0.50) and glutathione peroxidase (GPx) levels (3.41+/-0.09 to 6.17+/-0.65) increased significantly, pointing to hepatic oxidative stress. Altered, reduced and oxidized GSH levels showed significant recovery after MiADMSA and DMSA administration while, vitamins E and C were effective in reducing GSSG and TBARS levels and increasing catalase activity. Administration of MiADMSA alone and the combined administration of vitamin C along with DMSA and MiADMSA were most effective in increasing hepatic GSH levels to 4.88+/-0.14, 4.09+/-0.12 and 4.30+/-0.06, respectively. Hepatic catalase also reached near normal level in animals co-administered vitamin C with DMSA or MiADMSA (82.5+/-4.5 and 84.2+/-3.5, respectively). Combined treatments with vitamins and the thiol chelators were also able to effectively reduce lead induced decrease in renal catalase activity and increase in TBARS and GPx level. Combination therapy, however, was unable to provide an effective reversal in the altered parameters indicative of oxidative stress in different brain regions, except in catalase activity. The result also suggests a beneficial role of vitamin E when administered along with the thiol chelators (particularly with MiADMSA) in reducing body lead burden. Blood lead concentration was reduced from 13.3+/-0.11 in lead control to 0.3+/-0.01 in MiADMSA plus vitamin E-treated rats. Liver and kidney lead concentration also showed a most prominent decrease in MiADMSA plus vitamin E co-administered rats (5.29+/-0.16 to 0.63+/-0.02 and 14.1+/-0.21 to 1.51+/-0.13 in liver and kidney, respectively). These results thus suggest that vitamin C administration during chelation with DMSA/MiADMSA was significantly beneficial in reducing oxidative stress however, it had little or no additive effect on the depletion of lead compared with the effect of chelators alone. Thus, the co-administration of vitamin E during chelation treatment with DMSA or MiADMSA could be recommended for achieving optimum effects of chelation therapy. PMID- 12732453 TI - The effects of coffee on enzymes involved in metabolism of the dietary carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine in rats. AB - The effects of coffee on the metabolism and genotoxicity of the dietary carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) were investigated. Coffee diminished the bacterial mutagenicity of PhIP in the Ames reversion assay through inhibition of cytochrome P450 1A2 (CYP1A2), a key enzyme involved in the metabolic activation of PhIP. When given as part of the diet (0, 1 or 5% w/w) to male Fischer-344 rats for 2 weeks, coffee affected the expression of hepatic enzymes involved in PhIP metabolism. Coffee increased the expression of CYP1A2 by 16-fold in the 5% coffee-treated group, and approximately half of this inductive effect was attributed to caffeine. Coffee also increased the expression of enzymes involved in the detoxication of PhIP. A 2-fold increase in expression of glutathione S-transferase alpha was observed, UDP-glucuronosyl transferase (UGTs) activities of p-nitrophenol increased 2-fold, while N(2)-and N3-glucuronidation of the genotoxic metabolite 2-hydroxyamino-1-methyl-6 phenylimidazo[4,5-b]pyridine (HONH-PhIP) increased by 1.3-fold in the 5% coffee treated over the control group. The amount of PhIP (0.75 mg/kg, 24 h) eliminated in urine as the N(2)-and N3-glucuronide conjugates of HONH-PhIP increased by 1.8- and 2.5-fold, respectively, in the 5% coffee-treated group over control rats, suggesting either increased rates of N-oxidation of PhIP or N-glucuronidation of HONH-PhIP. Despite the strong induction of CYP1A2, there was no increase in PhIP DNA adduct formation in colon and pancreas while liver adducts decreased by 50% over control animals. These data suggest that the effect of coffee on inhibition of PhIP N-oxidation and ensuing DNA damage is more important in vivo than its effect on induction of PhIP N-hydroxylation. PMID- 12732455 TI - Molecular modelling of human CYP1B1 substrate interactions and investigation of allelic variant effects on metabolism. AB - Molecular modelling of human CYP1B1 based on homology with the mammalian P450, CYP2C5, of known three-dimensional structure is reported. The enzyme model has been used to investigate the likely mode of binding for selected CYP1B1 substrates, particularly with regard to the possible effects of allelic variants of CYP1B1 on metabolism. In general, it appears that the CYP1B1 model is consistent with known substrate selectivity for the enzyme, and the sites of metabolism can be rationalized in terms of specific contacts with key amino acid residues within the CYP1B1 heme locus. Furthermore, a mode of binding interaction for the inhibitor, alpha-naphthoflavone, is presented which accords with currently available information. The current paper shows that a combination of molecular modelling and experimental determinations on the substrate metabolism for CYP1B1 allelic variants can aid in the understanding of structure-function relationships within P450 enzymes. PMID- 12732456 TI - Increased susceptibility of renal epithelial cells to TNFalpha-induced apoptosis following treatment with fumonisin B1. AB - Previous studies have shown that tumor necrosis factor alpha (TNFalpha) is involved in the pathogenic events following exposure to fumonisin B(1) (FB(1)), a potent inhibitor of ceramide synthase and sphingolipid biosynthesis. The intimate role of sphingolipid mediators in TNFalpha signaling and cellular death suggests that FB(1) may alter the sensitivity of cells to TNFalpha-induced apoptosis. We tested the hypothesis that FB(1) treatment will increase the sensitivity of porcine renal epithelial cells to TNFalpha. Porcine renal epithelial cells (LLC PK(1)) were treated with FB(1) for 48 h prior to treatment with TNFalpha. A dose dependent increase in TNFalpha-induced apoptosis was observed in cells pretreated with FB(1). Cells treated with FB(1) showed increased DNA fragmentation and terminal uridine nucleotide end labeling in response to TNFalpha treatment. FB(1) increased DNA synthesis and resulted in cell cycle arrest in the G(2)/M phase of the cell cycle. Flow cytometric analysis of the cell cycle indicated that TNFalpha predominantly killed cells in the G(2)/M phase. The activation of JNK, a mitogen-activated protein kinase (MAPK), was increased following 48 h exposure to FB(1). Phosphorylation of p38 and ERK remained unchanged following treatment with FB(1). FB(1) also increased free sphingoid base levels under identical treatment conditions. Results suggest that FB(1) increased free sphingoid base levels and the population of cells in the G(2)/M phase. This population was shown to be most susceptible to TNFalpha-induced apoptosis. Phosphorylation of pro-apoptotic JNK may play an important role in these effects. PMID- 12732457 TI - Immunomodulation of human natural killer cell cytotoxic function by triazine and carbamate pesticides. AB - Triazine (atrazine) and carbamates (maneb, metiram, and ziram) are used as pesticides on a variety of crops around the world. To our knowledge, there have been no studies dealing with the effects of these compounds on human natural killer (NK) cells cytotoxic function. NK cells play a central role in immune defense against tumor development and viral infections. Thus, any agent that interferes with the ability of NK cells to lyse their targets could increase the risk of tumor incidence and/or viral infections. In this study, we examined the effects of atrazine, maneb, metiram, zineb, and ziram on the ability of human NK cells to lyse tumor cells. The compounds were tested in both purified NK cells as well as a cell preparation that contained both T and NK lymphocytes (T/NK cells). Lymphocytes were exposed to the compounds for periods of time ranging from 1 h to 6 days. Exposure of highly purified NK cells to 10 microM atrazine, maneb, or metiram inhibited K562 tumor cell lysis by 63+/-25, 95+/-4, and 50+/-6%, respectively, after a 24 h exposure and by 83+/-21, 70+/-39, and 48+/-41% after a 6-day exposure. Exposure to 2.5 microM ziram for 24 h caused a 99+/-2% decrease in lytic function and at 1 microM for 6 days caused a 96+/-4% decrease. However, when T/NK cells were exposed to atrazine, maneb, or metiram for 24 h only 10 microM atrazine and maneb caused a significant decreases in lytic function (61+/ 13 and 38+/-18%) and after 6 days only atrazine was inhibitory (54+/-12%). A 24-h exposure to 2.5-microM ziram caused a 41+/-51% decrease in function, but a 6-day exposure to 1 microM ziram caused no inhibition of lytic function. The results provide evidence of relative toxic potential for the five compounds and the immunomodulatory effects on both T and NK lymphocyte function. PMID- 12732458 TI - Increased acetylcholinesterase activities in specimens of Sparus auratus exposed to sublethal copper concentrations. AB - The present study looks at possible changes in the activity of acetylcholinesterase (AChE) in tissues (brain and white muscle) of the Mediterranean bony fish Sparus auratus after a 20 days exposure to sublethal concentrations (0.1 or 0.5 ppm) of copper in the marine water and on control untreated animals. The trials also included measurements of Cu concentration in the tissues to evaluate possible metal accumulation. Moreover, sedimentation analysis as well as V(max) and K(m) determination were carried out in tissue extracts of Cu-exposed or control animals. V(max) and K(m) were also determined with or without addition of Cu(2+) in the assay. No Cu accumulation occurred in brain and muscle after Cu exposure. AChE showed in both tissues a molecular polymorphism with putative globular (G) and asymmetric (A) forms. Cu exposition led to an increased specific activity and improved catalytic efficiency of AChE in brain and muscle, seemingly regarding G forms. The increase in catalytic efficiency also resulted from the in vitro assay with tissue extracts and Cu(2+) addition. The higher AChE activity and catalytic efficiency in both tissues after Cu exposition and without metal accumulation, suggests an increase of free Cu aliquot into the cells, likely due to mechanisms of metal homeostasis. PMID- 12732460 TI - Role of intracellular reactive oxygen species and mitochondrial dysfunction in evening primrose extract-induced apoptosis in Ehrlich ascites tumor cells. AB - Herbal medicines are increasingly being utilized to treat a wide variety of disease processes. Evening primrose extract (EPE) is extracted from Oenothera biennis L., one species of evening primroses, which has been shown to have several pharmacological effects. However, anti-tumor activity in the extract of defatted seeds of O. biennis L. has not been defined thus far. In this study, we identified the major biochemical changes upon EPE treatment and investigated the functional relationship between these changes. We found that EPE-induced apoptosis in Ehrlich ascites tumor cells as evidenced by morphological changes. Furthermore, our results demonstrated rapid increase of intracellular peroxides levels, loss of mitochondrial membrane potential and the release of cytochrome c from mitochondria to cytosol. These results suggest that the rapid increase of intracellular peroxides levels after addition of EPE triggers off induction of apoptosis. PMID- 12732459 TI - In vitro action of combinations of selected antimicrobial agents and adult bovine articular cartilage (sesamoid bone). AB - Anatomically intact articular cartilage in form of sesamoid bones from metacarpophalangeal joints of 2-year-old cows was tested for its influence on the microbicidal effect of the iodophore Betaisodona, the bispyridinamine Octenisept, and the biguanide Lavasept. Comparisons were carried out in Ham's F12 medium with and without 0.2% bovine serum albumin as organic matter loading. The expected abolition of the microbicidal effect of these antiseptics against the test organisms Escherichia coli or Staphylococcus aureus in the presence of sesamoid bone was not evident. Furthermore, sesamoid bone alone demonstrated antibacterial activity against Staphylococcus aureus, which may involve adherence of bacteria to surface constituents of articular cartilage. Final concentrations of 2.5-5% Betaisodona, 5% Octenisept as well as 0.025% Lavasept are effective in killing of 10(8)-10(9) cfu/ml Escherichia coli or Staphylococcus aureus in the presence of sesamoid bone without the reduction of antimicrobial activity expected from binding to CS, which has previously been demonstrated for CS in solution. PMID- 12732461 TI - New findings in the study on the intercalation of bisdaunorubicin and its monomeric analogues with naked and nucleus DNA. AB - DNA is a target molecule for anthracycline anticancer drugs. We have used new anthracycline derivatives, bisdaunorubicin (WP631) and its monomeric analogues (WP700 serie), and look if there was a relation between the drug binding affinity to naked DNA and to cell nucleus in the cell with its cytotoxicity. Circular dichroism (CD) and fluorescence were used to follow the interaction of anthracycline derivatives with naked DNA and cell nuclei. WP631 interacts with DNA at two distinct stoichiometries, 6:1 and 3:1 base pair (bp)/WP631 molecule (3:1 and 1.5:1 per anthracycline rings). Monomeric daunorubicin (DNR) with its amino sugar N-bound to amino- and nitro-substituted benzyl moiety, representing p xylenyl linker present in WP631 bisintercalator, is much more binding to DNA than DNR or WP631. These findings are supported by the study of drug binding by nuclei of K562 cells. Around 70% of WP700 intercalate to nucleus DNA in the steady state, while only 45% of DNR intercalate DNA in the cell. The binding of WP631 by K562 cells is even less effective ( approximately 20%). WP 700 compounds, which are very similar to each other in their binding to DNA, self-association and cell accumulation, differ very distinctly in their cytotoxicity power. The most effective compounds are amino-benzyl derivatives of WP 700 series. The nitro benzyl compounds have very low toxicity, even if they bind to DNA with similar power with that of the amino derivatives. The comparison of the all data clearly indicates no relation between cytotoxicity of the drug and its ability to intercalate DNA. PMID- 12732462 TI - A comparison of larval development and mucosal mast cell responses in worm-naive goat yearlings, kids and lambs undergoing primary and secondary challenge with Teladorsagia circumcincta. AB - Larval development, mucosal mast cell (MMC) and eosinophil responses in worm nai;ve lambs, yearling goats and goat kids were compared using two different experimental challenge regimes involving oral administration of infective Teladorsagia circumcincta L(3). Experimental challenge regimes enabled primary and secondary immune responses in the two species to be compared. Goats carried higher worm burdens than lambs and there were significant differences in the stages of development attained by the larval challenge that established in the two species. Possible physiological reasons for these differences are discussed. There were also differences in the establishment and development of larvae in individual yearlings which may indicate the development of a weak age-related immune response. Quantitative analysis of MMC and globule leukocyte (GL) recruitment and functional activity in the form of mast cell-specific proteinase (MCP) production demonstrated differences between the species with goat tissues containing significantly higher numbers of GL and lower concentrations of MCP than the lambs. Quantitative analysis of blood and tissue eosinophil responses failed to demonstrate any significant differences in either species under the two challenge regimes. PMID- 12732463 TI - Regulation of the resistance to nematode parasites of single- and twin-bearing Merino ewes through nutrition and genetic selection. AB - Periparturient Merino ewes obtained from lines of sheep that had been selected either for increased resistance to Haemonchus contortus (R) or at random (C) were supplemented, while grazing at pasture, with either nil or 250 g/day cottonseed meal (CSM) for the 6 weeks prior to or the 6 weeks after the start of parturition. Ewes from both supplement groups had lower (mean 66% reduction) faecal egg counts (FECs) during the postpartum period and this coincided with a period of maternal body weight loss. Factors which increased the rate of maternal body weight loss, such as pregnancy and lactation status, also increased FEC. Evidence is presented that the magnitude of the periparturient rise (PPR) in FEC in grazing ewes will be greatest during periods of maternal weight loss and at these times supplementation to increase metabolisable protein (MP) supply will be most effective in increasing resistance to nematode parasites. The resistance of R ewes to nematode parasites was greater than that of C ewes throughout the experiment and was sufficiently low such that anthelmintic treatment in a commercial environment may not have been required. Irrespective of actual FEC, ewes from all treatment combinations exhibited a PPR in FEC. Reduced FEC of R ewes resulted in reduced apparent pasture larval contamination after 18 weeks of continuous grazing but supplementation was ineffective in this regard. It is suggested that integrated parasite management (IPM) programs for periparturient ewes should make use of both protein supplementation and genetic selection to increase worm resistance and reduce dependency on anthelmintics for worm control. PMID- 12732464 TI - Prevalence of benzimidazole resistant nematodes in sheep flocks in Yucatan, Mexico. AB - Thirty-eight sheep flocks, located in three municipalities in the Eastern Yucatan, Mexico, were surveyed for gastrointestinal nematodes resistant to benzimidazole (BZD) anthelmintics (AH). On each flock, 30 sheep were randomly distributed into two groups of 15 animals: albendazole group (5mg/kg BW) and untreated control group. Animals were refrained from any food (either browsing/grazing or supplement) for a period of 16 h prior to treatment. Faecal egg counts (FEC) and larval cultures were performed 10 days after anthelmintic treatment. Percentage reduction and 95% confidence intervals were determined. Anthelmintic resistance (AR) was declared when the percentage reduction in FEC was <95% and the 95% confidence interval was <90%. AR was suspected when only one of the two criteria was met. The survey indicated that AR occurred in 15.8% (n=6) (95% confidence interval=+/-11.6%) and was suspected in 23.7% of the farms (n=9) (95% confidence interval=+/-13.3%). Post-treatment larval cultures indicated that Haemonchus was the only resistant genus. The questionnaire survey showed that most farmers (92%) considered their sheep a secondary activity to cattle production. The majority of farmers (97.4%) treat their animals according to visual appreciation of weight. However, most farmers (79%) treat their flocks at very low frequencies (<3 times per year). Drug rotation was performed every 12 months or more by 84.2% of farmers. Anthelmintics used were: macrociclic lactones (47.4%), BZD (39.5%), levamisol (10.5%) and 1 farmer used closantel. PMID- 12732465 TI - The prevalence of Toxocara cati in domestic cats in Mexico City. AB - The faeces of 520 domestic cats, resident in the 16 municipal authorities of Mexico City, were analysed using the method of centrifuging with zinc sulphate (at 33%). Three hundred and ninety-nine animals lived in houses, 121 in apartments. For the purpose of this study they were divided into seven age groups, of 6 months each. Toxocara cati eggs were found in the faeces of 42.5% of the animals. 20.7% of apartment cats and 49.1% of house cats were infected. This difference in the rate of infection is statistically significant (P<0.002.) for the house cats. T. cati was found in all the age groups although most infection was found in animals less than 1 year old. The fact that infection occurred in the whole age range of the cats and in all the municipal authorities means that the risk of being contaminated with this helminth in Mexico City is high. As a consequence, the possibility of developing illness as a result of becoming a host to larva migrans is also high. PMID- 12732466 TI - Worm control practices and anthelmintic usage in traditional and dairy cattle farms in the southern highlands of Tanzania. AB - Worm control practices and anthelmintic usage in 177 cattle farms in Iringa district in the southern highlands of Tanzania was determined through a questionnaire survey. A total of 76 traditional, 92 small-scale dairy and 9 large scale dairy cattle farms were included in the survey. Results indicated that 87.7% traditional, 97.8% small-scale dairy and 100% large-scale farmers relied solely on the use of anthelmintics, 2.7% traditional farmers used traditional medicines while 9.6% traditional farmers had not any form of worm control practice. Worm infection was ranked the second most important constraint of productivity in cattle in the three production systems. Most farms (57.6% traditional, 35.8% small-scale dairy, 66.7% large-scale dairy) used anthelmintics with a combination of levamisole and oxyclozanide. Benzimidazoles were used only in traditional (25.4%) and small-scale dairy (32.1%) farms while nitroxynil (Trodax) was mostly used in large-scale dairy farms (33.3%). Generally, 40% of farmers treated three or four times a year and the frequency in some farms was surprisingly high for resource poor small-scale farmers. The frequency of anthelmintic treatment was mostly the same regardless of the management system. Treatments in most farms depended on availability of money and drugs and not the epidemiology of parasites. A significant proportion (46.3%, P=0.007) of farmers especially in rural areas failed to follow their pre-planned treatment schedules due to lack of money (86%) and unavailability of drugs (6.6%). Many farmers (58.9%) had used the same type of anthelmintic for four or more consecutive years and 85.3% of them would continue with the same anthelmintic. Farmers in all management systems mostly purchased anthelmintics from private veterinary drug shops and about 43% traditional and 33.3% small-scale dairy farmers mostly in rural areas obtained anthelmintics from village extension officers. Despite the fact that all farmers were aware of worm infection and the associated signs in cattle, 42.5% had poor knowledge on the source of worm infection. Small-scale dairy farmers allowed only a 1-day withdraw period for milk regardless of the type of anthelmintic used and there was no milk and slaughter clearance in traditional farms. It was concluded from this study that worm control in Iringa faces serious constrains and that education of farmers and farm hands is not adequate. Moreover, poor quality control and high price of potent anthelmintics, few extension workers, low income and low education among farmers contributed significantly to erratic worm control practices and anthelmintic usage in peri urban and rural areas. PMID- 12732467 TI - Detection of antibodies in sera of weaned pigs after contact infection with Sarcoptes scabiei var. suis and after treatment with an antiparasitic agent by three different indirect ELISAs. AB - Three commercial enzyme-linked immunosorbent assays (ELISAs) were compared for the detection of antibodies to Sarcoptes scabiei var. suis using experimental sera of six 8-week-old pigs after contact infection with Sarcoptes scabiei var. suis. Six non-infected pigs were monitored as a control group. Blood sera were taken once a week from all animals. After successful infection the pigs were treated with an antiparasitic agent (12 weeks post infection (p.i.)) and the antibody titres were monitored until they were negative. The antibody levels of the experimental pigs reached the cut-off level 5 weeks after introduction of an infected animal to the group and were positive by both the Sarcoptes-ELISA 2001 PIG and the Acar-Test P-ELISA. Four weeks after treatment mean results showed optical densities (% OD) below the cut-off level in the Sarcoptes-ELISA 2001 and 8 weeks after treatment in the Acar-Test P-ELISA. In the Chekit Sarcoptest pigs had elevated antibody levels in comparison to control animals, but ODs remained below the given cut-off level at all times. In a second examination with Chekit Sarcoptest (different lot) and at a lower cut-off level, the sera of most of the piglets tested positive. Eight weeks after treatment, four from six pigs still had positive OD values. Therefore this investigation showed a higher sensitivity for the Sarcoptes-ELISA 2001 and the Acar-Test P-ELISA than for the Chekit Sarcoptest. Different test sensitivities must be considered when serologic methods are used for the diagnosis of swine sarcoptic mange, especially for monitoring and controlling eradication programs. PMID- 12732468 TI - Nodular onchocercosis of red deer in central Spain. AB - The presence of nodular onchocercosis was investigated post-mortem in 142 red deer (11 calves, 35 yearlings and 96 adults) shot from February 1998 to January 1999, and July-November 1999 in "Quintos de Mora" (Toledo, central Spain), a game property belonging to the National Wildlife Reserves. Between 6 and 13 animals were analysed monthly by inspection for subcutaneous nodules of Onchocerca spp. Subcutaneous nodules of Onchocerca sp. were detected in 24% of the animals. Anatomical locations for nodules were the back and flanks. Infection ranged from 1 to 29 nodules per animal. Adult worms collected from nodules were identified as O. flexuosa. No apparent seasonal trend was observed either in prevalence or in mean intensity of infection, which fluctuated between 48% (5+/-8 nodules) in winter 1998 and 5% (1+/-0 nodules) in summer 1999. Prevalence of infection was significantly higher in adult (30%) than in young animals (9% in calves, 11% in yearlings), although no age effect on intensity was observed. The size of the nodules was measured to evaluate the age of infection. Small (recent) nodules (5 6mm of diameter) were collected in late spring, summer and fall; medium-sized nodules (12-15 mm) were in second half of summer, fall and winter, and large (mature) nodules (20-25 mm) in fall, winter and part of spring. Significant differences were found among host age groups. PMID- 12732469 TI - Contribution of Efa1/LifA to the adherence of enteropathogenic Escherichia coli to epithelial cells. AB - Enteropathogenic E. coli(EPEC) is an important diarrhoeal pathogen that induces characteristic lesions on the host intestine termed attaching and effacing (A/E) lesions. In this study we have examined the contribution of a large gene, efa1, which is present in all A/E pathogens, to the adherence phenotype of EPEC. An efa derivative of EPEC JPN15 was constructed and this mutant was significantly less adherent to epithelial cells than the parent strain. The JPN15 efa- derivative was FAS-positive, produced EspA filaments and showed comparable levels of EspA secretion to JPN15. In addition, polyclonal antibodies raised to Efa1 partially inhibited the adherence of JPN15 to cultured epithelial cells. In further work, we showed that human and rabbit hosts infected with an A/E pathogen produced antibodies to Efa1 and we observed that the truncated form of efa1 present in EHEC O157:H7 was specific to that serotype. Generally efa1 was present in its entirety in the genomes of other A/E pathogens. Overall our data suggest that Efa1 has host cell binding activity, at least in tissue culture, and that it is produced during infection. These findings suggest that Efa1 may play a direct role in the pathogenesis of infections caused by A/E pathogens. PMID- 12732470 TI - Biological effects of two genetically defined leukotoxin mutants of Mannheimia haemolytica. AB - Mannheimia(Pasteurella)haemolytica serotype 1 is the primary causative agent responsible for bovine pneumonic mannheimiosis, also known as shipping fever in cattle. The bacterium produces a variety of virulence factors, foremost of which is the exotoxic leukotoxin. The leukotoxin is a calcium-dependent cytolysin that is a member of the RTX (repeats in toxin) family and exhibits a narrow cell-type and species specificity and has biological effects only on ruminant leukocytes and platelets. The genetic organization of the leukotoxin is comprised of four genes: lktC, lktA, lktB and lktD. The lktA structural gene encodes the protoxin (pro-LktA) and lktC encodes a transacylase that post-translationally modifies the inactive pro-LktA to a biologically active wild-type leukotoxin (LktA). The LktA has been implicated as the key factor that contributes to the pathogenesis of lung injury associated with the disease and considerable efforts have been employed in abrogating toxin function while retaining immunogenicity, with an eye towards design of attenuated vaccines. We hypothesized that the pro-LktA retains the ability to cause biological effects on target cells as has been reported in the case of the closely related RTX toxin alpha-hemolysin (HlyA). We also examined the biological effects of an amino-terminal truncation mutant leukotoxin DeltaLktA on target cells. Thus the objectives of our study were to investigate whether two different mutant leukotoxins, one a nonacylated pro-LktA, and the other lacking 344 amino acids at the N-terminal end of the LktA protein; DeltaLktA, are capable of (i). binding to the beta2-integrin leukotoxin receptor, (ii). inducing the elevation of second messenger intracellular calcium ([Ca(2+)](i)), and (iii). inducing inflammatory gene expression, reactive oxygen metabolites (ROMs) and cytolysis in target cells. Our results demonstrate that neither acylation nor the amino terminal 344 amino acids are required for LktA binding but are essential for LktA-induced [Ca(2+)](i) elevation, generation of ROM, generation of the inflammatory cytokine IL-8 and cytolysis in target cells. PMID- 12732471 TI - Extracellular secretion of the virulence plasmid-encoded ADP-ribosyltransferase SpvB in Salmonella. AB - Nontyphoid Salmonella enterica requires the plasmid-encoded spv genes to establish successful systemic infection in experimental animals. The SpvB virulence-associated protein has recently been shown to contain the ADP ribosyltransferase domain. SpvB ADP-ribosilates actin and depolymerizes actin filaments when expressed in cultured epithelial cells. However, spontaneous secretion or release of SpvB has not been observed under in vitro growth conditions. In the present study we investigated the secretion of SpvB from Salmonella using in vitro and in vivo assay systems. We showed that SpvB is secreted into supernatant from Salmonella strains that contain the cloned spvB gene on a plasmid when they grew in intracellular salts medium (ISM), a minimal medium mimicing the intracellular iron concentrations of eukaryotic cells. A series of mutant SpvB proteins revealed that an N-terminal region of SpvB located at amino acids 1-229 was sufficient to promote secretion into extracellular milieu. Confocal immunofluorescence microscopy also demonstrated efficient localization of the N-terminal domain of SpvB(1-360) tagged with biotinylated peptide within infected host cell cytosol but not truncated SpvB(1-179) fusion protein. In addition, mutations that inactivate genes within Salmonella pathogenicity island 1 or Salmonella pathogenicity island 2 that encode type III secretion systems (TTSS) could secrete the SpvB protein into the culture medium. These results indicate that SpvB protein is transported from the bacteria and into the host cytoplasm independent of TTSS. PMID- 12732472 TI - Experimental tularemia in mice challenged by aerosol or intradermally with virulent strains of Francisella tularensis: bacteriologic and histopathologic studies. AB - BALB/c and C57BL/6 mice were challenged by aerosol or intradermally with low doses ( approximately 10-20 colony forming units) of virulent type A and type B strains of the facultative intracellular pathogen, Francisella tularensis, and the course of infection was monitored. Both mouse strains were equally susceptible to infection, but type A strains reached lethal numbers a few days earlier than type B strains regardless of challenge route. BALB/c mice showed overt signs of infection for several days, whereas C57BL/6 mice remained asymptomatic until a few hours before death. Histological changes were extensive and severe in the liver and spleen, but much more limited in the lungs, even in mice challenged by aerosol. Thus, it appears that regardless of the route of infection, systemic rather than pulmonary infection was the likely cause of death following low dose challenge with virulent F. tularensis. PMID- 12732474 TI - Effective size in management and conservation of subdivided populations. AB - A numerical method for computing the eigenvalue variance effective size of a subdivided population connected by any fixed pattern of migration is described. Using specific examples it is shown that total effective size of a subdivided population can become less than the sum of the subpopulation sizes as a result of directionalities in the pattern of migration. For an extension of the model with threshold harvesting and local deterministic logistic population dynamic we consider the problem of maximizing the total harvesting yield with constraints on the total effective size. For some simple source-sink systems and more complicated population structures where subpopulations differ in their degree of isolation, it is shown to be optimal, for a given total effective size, to raise the harvesting thresholds relatively more in small and in isolated populations. Finally, we show how the method applies to populations which are supplemented, either intentionally or unintentionally. It is shown that the total effective size can be reduced by several orders of magnitude if the captive component of a population is much smaller than the wild component, even with symmetric backward migration. PMID- 12732473 TI - A polar flagella operon (flg) of Aeromonas hydrophila contains genes required for lateral flagella expression. AB - Aeromonas spp. are pathogens of both humans and poikilothermic animals, causing a variety of diseases. Certain strains are able to produce two distinct types of flagella; polar flagella for swimming in liquid and lateral flagella for swarming over surfaces. Although, both types of flagella have been associated as colonisation factors, little is known about their organisation and expression. Here we characterised a complete flagellar locus of Aeromonas hydrophila (flg) containing 16 genes, this was analogous to region 1 of the Vibrio parahaemolyticus polar flagellum, with the difference that no flagellin genes were found on A. hydrophila while V. parahaemolyticus showed three flagellin genes. The flg region was present in all Aeromonas strain tested. Defined insertion mutants in flgL, were unable to swim, had a drastic reduction in swarming, lateral flagella, HEp-2 cell adhesion and biofilm formation. Mutations in flgN caused a drastic reduction in lateral flagella, inability to swarm, but these strains were still able to swim. Whereas the cheV mutants still produced both types of flagella and were able to swim and swarm. These results suggest that FlgN is required for lateral flagella formation and swarming motility, but not for polar flagellum-mediated swimming. PMID- 12732475 TI - Size-dependent ESS sex allocation in wind-pollinated cosexual plants: fecundity vs. stature effects. AB - To theoretically investigate the single and compound effects of relative fecundity and relative stature of plants on size-dependent sex allocation (SDS) in wind-pollinated cosexual species, we developed a game model and analysed ESS sex allocation of large and small plants having totally or partially different reproductive resources and different pollen and seed dispersal areas in a population. We found that e.g. when both sized plants have large pollen dispersal areas relative to their seed dispersal areas, which plants are male-biased is largely determined by relative fecundity (t) and relative size of seed dispersal area (k) of the large plants to the small plants: If t >k, large plants tend to be more male-biased even if relative size of pollen dispersal area of large to small plants (l) is smaller than k. If t 0.05) for analysis of variance and Student-Newman-Keuls analysis, although the 95% confidence interval (CI) of grade I was different from grade II. Every degree of reflux volume showed close correlation with the reflux index. Other indices were less sensitive. CONCLUSIONS: Venous reflux volume is a more accurate non-invasive quantitative assessment of the degree of deep venous reflux in the lower extremities than current methods. PMID- 12732496 TI - Recurrent respiratory papillomatosis. AB - A 6-year-review of patients who presented with recurrent respiratory papillomatosis (RRP) to our hospital from January 1996 to December 2001 was carried out. Ten cases were identified, of which six were juvenile-onset RRP. Hoarseness was the most common symptom, noted in nine (90%) patients. Other clinical presentations included cough, stridor and aphonia. All patients had glottic papillomas; two had multiple sites of involvement. One patient underwent a tracheostomy that revealed papillomas over the trachea, bronchus and lung parenchyma. Half of the patients were Chinese. Of the six cases of juvenile-onset RRP, three patients were Malay, two Chinese and one Indian. Three Chinese and one German patient had adult-onset RRP. Among the juvenile-onset RRP cases, the mean age at presentation was 2 years, while for adult-onset RRP, it was 42 years. Juvenile-onset RRP was more common in females. There were more papillomas over more sites in patients with juvenile-onset RRP than with adult-onset disease. Subglottic involvement was noted in the juvenile-onset RRP cases. All patients were treated with CO2 laser therapy, but there was complete remission of the papillomas in only two cases. PMID- 12732497 TI - Deployment of endograft in the ascending aorta to reverse type A aortic dissection. AB - Current surgery to treat acute type A aortic dissection involving an intimal tear in the ascending aorta consists of resection and replacement, but mortality is high. We report the case of a 46-year-old female patient with Marfan syndrome who presented with excruciating retrosternal pain and breathing distress after a bowel movement with stress. Magnetic resonance imaging and multicolour sonography showed type A aortic dissection extending from the aortic root to the right iliac artery, with intimal tears in the ascending aorta above the sinotubular junction, the distal arch beyond the left subclavian artery and the isthmic region. We adapted the endoluminal stenting technique to this case of type A aortic dissection by sealing the intimal tears in the ascending aorta using endovascular introduction of one endoluminal graft, as confirmed on angiography. The patient was discharged after 10 days. Follow-up examination by computed tomography after more than 1 year revealed no sign of dissection at any level of the aorta. PMID- 12732498 TI - Molecular genetic analysis of anaplastic pleomorphic xanthoastrocytoma. AB - A case of pleomorphic xanthoastrocytoma in a 10-year-old Malay boy is reported. The patient presented with headache and epilepsy. On computed tomography, a ring enhancing low-density lesion was observed in the left fronto-temporal area. During surgery, a cystic tumour containing serous fluid was found and almost totally removed. Histologically, the tumour exhibited marked pleomorphism of oval and spindle-shaped cells intermixed with uni- and multinucleated giant cells, and xanthomatous cells with foamy cytoplasm. The tumour displayed pericellular reticulin and periodic acid-Schiff positive granules. Focally, six mitotic characters per 10 high-power fields were seen, and necrosis was confined only to the inner lining of the cyst. Mutational analysis showed that a frameshift mutation (a 4-bp deletion) in the p53 gene had occurred in codons 273 and 274 of exon 8. No mutation was detected in the p16 gene. No allelic loss and/or loss of heterozygosity were observed on chromosome 10 using microsatellite marker D105532. The patient was treated with postoperative radiotherapy because of histological anaplasia and the presence of residual tumour. The patient showed marked neurological recovery after a follow-up period of 2 years. PMID- 12732499 TI - Penile fracture with complete urethral rupture. AB - We report a rare case of penile fracture with complete urethral rupture in a 25 year-old male who sustained the injury during sexual intercourse. He presented with a tense haematoma on the ventral aspect of the penile shaft, associated with per urethral bleeding. Despite the injury, he was able to void painfully. Retrograde urethrography revealed complete obstruction at the proximal third of the urethra. Exploration and repair of the penile fracture and urethra were performed. The patient made an uneventful recovery with good erectile and voiding function. This case illustrates the value of retrograde urethrography in assessing urethral injuries in patients with penile fracture. PMID- 12732500 TI - Virtual reality and 3D visualizations in heart surgery education. AB - BACKGROUND: Computer assisted teaching plays an increasing role in surgical education. The presented paper describes the development of virtual reality (VR) and 3D visualizations for educational purposes concerning aortocoronary bypass grafting and their prototypical implementation into a database-driven and internet-based educational system in heart surgery. MATERIALS AND METHODS: A multimedia storyboard has been written and digital video has been encoded. Understanding of these videos was not always satisfying; therefore, additional 3D and VR visualizations have been modelled as VRML, QuickTime, QuickTime Virtual Reality and MPEG-1 applications. An authoring process in terms of integration and orchestration of different multimedia components to educational units has been started. RESULTS: A virtual model of the heart has been designed. It is highly interactive and the user is able to rotate it, move it, zoom in for details or even fly through. It can be explored during the cardiac cycle and a transparency mode demonstrates coronary arteries, movement of the heart valves, and simultaneous blood-flow. Myocardial ischemia and the effect of an IMA-Graft on myocardial perfusion is simulated. Coronary artery stenoses and bypass-grafts can be interactively added. 3D models of anastomotique techniques and closed thrombendarterectomy have been developed. Different visualizations have been prototypically implemented into a teaching application about operative techniques. CONCLUSIONS: Interactive virtual reality and 3D teaching applications can be used and distributed via the World Wide Web and have the power to describe surgical anatomy and principles of surgical techniques, where temporal and spatial events play an important role, in a way superior to traditional teaching methods. PMID- 12732501 TI - Unrelated donor stem cell transplantation compared with chemotherapy for children with acute lymphoblastic leukemia in a second remission: a matched-pair analysis. AB - Allogeneic stem cell transplantation (SCT) is frequently considered as treatment for relapsed childhood acute lymphoblastic leukemia (ALL). For patients without a matched sibling donor, SCT from unrelated donors (UD-SCT) has been increasingly performed during the past years. However, UD-SCT-related mortality and morbidity is still considerable, and the question remains as to which patients are at such high risk of recurrence that UD-SCT is indicated and, conversely, which patients do not require transplantation for long-term disease control. A matched-pair analysis was performed among patients treated according to Acute Lymphoblastic Leukemia Relapse Berlin-Frankfurt-Munster (ALL-REZ BFM) Study Group protocols after first relapse with chemotherapy or UD-SCT. Altogether 81 pairs were identified that could be matched exactly for site of relapse and immunophenotype, and as closely as possible for duration of first remission, age, diagnosis date, and peripheral blast cell count at relapse. No significant difference in the probability of event-free survival (pEFS) between UD-SCT and chemotherapy existed regarding 28 pairs with an intermediate prognosis (0.39 +/- 0.10 vs 0.49 +/- 0.11, P =.105), whereas the pEFS was significantly different in the 53 pairs with a poor prognosis (0.44 +/- 0.07 vs 0.00 +/- 0.00, P <.001). The major reasons of treatment failure among patients who underwent UD-SCT were therapy-related death (TRD; 24/81) and relapses (20/81). In contrast, TRD rarely occurred in patients treated with chemotherapy alone (3/81), but relapse was much more common (62/81). In conclusion, UD-SCT provides better event-free survival for children with high risk relapsed ALL. However, there is no clear advantage of UD-SCT in patients with intermediate prognosis. PMID- 12732502 TI - Megakaryocytes require thrombospondin-2 for normal platelet formation and function. AB - Mice that lack the matricellular angiogenesis inhibitor, thrombospondin-2 (TSP2), display a bleeding diathesis, despite normal blood coagulation and the lack of thrombocytopenia. Although platelets do not contain detectable levels of TSP2, TSP2-null platelets are compromised in their ability to aggregate in vivo in response to denudation of the carotid artery endothelium, and in vitro following exposure to adenosine diphosphate (ADP). Megakaryocytes (MKs) show high levels of TSP2 by immunohistochemical analysis of bone marrow. However, when cultured in vitro, MKs contain little TSP2 protein or mRNA. These findings suggest that most TSP2 is acquired from the bone marrow microenvironment. Consistent with this hypothesis, MKs take up recombinant TSP2 in an integrin-dependent manner when it is supplied in the culture medium. Furthermore, uptake of TSP2 in vitro affects MK differentiation and proplatelet formation. The functional significance of this process is supported by the presence of ultrastructural abnormalities in TSP2 null bone marrow, including extensive fragmentation of the peripheral zone in MKs and failure of this zone to form close associations with vascular sinuses. We conclude that the uptake of TSP2 by MKs from the marrow milieu is required for proper MK function and the release of functionally competent platelets. PMID- 12732504 TI - The -1C>T mutation in the annexin A5 gene does not affect plasma levels of annexin A5. PMID- 12732503 TI - Reassessment of loss of heterozygosity within MLL in childhood acute lymphoblastic leukemia. PMID- 12732505 TI - Agranulocytosis unresponsive to growth factors following rituximab in vivo purging. PMID- 12732506 TI - Cytolytic toxin Cyt1A and its mechanism of membrane damage: data and hypotheses. PMID- 12732507 TI - Quantitative detection of methanotrophs in soil by novel pmoA-targeted real-time PCR assays. AB - Methane oxidation in soils is mostly accomplished by methanotrophic bacteria. Little is known about the abundance of methanotrophs in soils, since quantification by cultivation and microscopic techniques is cumbersome. Comparison of 16S ribosomal DNA and pmoA (alpha subunit of the particulate methane monooxygenase) phylogenetic trees showed good correlation and revealed five distinct groups of methanotrophs within the alpha and gamma subclasses of Proteobacteria: the Methylococcus group, the Methylobacter/Methylosarcina group, the Methylosinus group, the Methylocapsa group, and the forest clones group (a cluster of pmoA sequences retrieved from forest soils). We developed quantitative real-time PCR assays with SybrGreen for each of these five groups and for all methanotrophic bacteria by targeting the pmoA gene. Detection limits were between 10(1) and 10(2) target molecules per reaction for all assays. Real-time PCR analysis of soil samples spiked with cells of Methylococcus capsulatus, Methylomicrobium album, and Methylosinus trichosporium recovered almost all the added bacteria. Only the Methylosinus-specific assay recovered only 20% of added cells, possibly due to a lower lysis efficiency of type II methanotrophs. Analysis of the methanotrophic community structure in a flooded rice field soil showed (5.0 +/- 1.4) x 10(6) pmoA molecules g(-1) for all methanotrophs. The Methylosinus group was predominant (2.7 x 10(6) +/- 1.1 x 10(6) target molecules g(-1)). In addition, bacteria of the Methylobacter/Methylosarcina group were abundant (2.0 x 10(6) +/- 0.9 x 10(6) target molecules g of soil(-1)). On the other hand, pmoA affiliated with the forest clones and the Methylocapsa group was below the detection limit of 1.9 x 10(4) target molecules g of soil(-1). Our results showed that pmoA-targeted real-time PCR allowed fast and sensitive quantification of the five major groups of methanotrophs in soil. This approach will thus be useful for quantitative analysis of the community structure of methanotrophs in nature. PMID- 12732508 TI - Clade-specific 16S ribosomal DNA oligonucleotides reveal the predominance of a single marine Synechococcus clade throughout a stratified water column in the Red Sea. AB - Phylogenetic relationships among members of the marine Synechococcus genus were determined following sequencing of the 16S ribosomal DNA (rDNA) from 31 novel cultured isolates from the Red Sea and several other oceanic environments. This revealed a large genetic diversity within the marine Synechococcus cluster consistent with earlier work but also identified three novel clades not previously recognized. Phylogenetic analyses showed one clade, containing halotolerant isolates lacking phycoerythrin (PE) and including strains capable, or not, of utilizing nitrate as the sole N source, which clustered within the MC A (Synechococcus subcluster 5.1) lineage. Two copies of the 16S rRNA gene are present in marine Synechococcus genomes, and cloning and sequencing of these copies from Synechococcus sp. strain WH 7803 and genomic information from Synechococcus sp. strain WH 8102 reveal these to be identical. Based on the 16S rDNA sequence information, clade-specific oligonucleotides for the marine Synechococcus genus were designed and their specificity was optimized. Using dot blot hybridization technology, these probes were used to determine the in situ community structure of marine Synechococcus populations in the Red Sea at the time of a Synechococcus maximum during April 1999. A predominance of genotypes representative of a single clade was found, and these genotypes were common among strains isolated into culture. Conversely, strains lacking PE, which were also relatively easily isolated into culture, represented only a minor component of the Synechococcus population. Genotypes corresponding to well-studied laboratory strains also appeared to be poorly represented in this stratified water column in the Red Sea. PMID- 12732509 TI - Concentration and prevalence of Escherichia coli O157 in cattle feces at slaughter. AB - The concentration and prevalence of Escherichia coli O157 in cattle feces at the time of slaughter was studied over a 9-week period from May to July 2002. Fecal samples (n = 589) were collected from the rectums of slaughtered cattle, and the animal-level prevalence rate was estimated to be 7.5% (95% confidence interval [CI], 5.4 to 9.6%) while the group prevalence was 40.4% (95% CI, 27.7 to 53.2%). Of the 44 infected animals detected, 9% were high shedders that contained E. coli O157 at concentrations of >10(4) CFU g(-1). These 9% represented >96% of the total E. coli O157 produced by all animals tested. All isolates possessed the vt(2) gene, 39 had the eaeA gene, and a further five had the vt(1) gene also. The presence of high-shedding animals at the abattoir increases the potential risk of meat contamination during the slaughtering process and stresses the need for correctly implemented hazard analysis and critical control point procedures. PMID- 12732510 TI - Prokaryotic metabolic activity and community structure in Antarctic continental shelf sediments. AB - The prokaryote community activity and structural characteristics within marine sediment sampled across a continental shelf area located off eastern Antarctica (66 degrees S, 143 degrees E; depth range, 709 to 964 m) were studied. Correlations were found between microbial biomass and aminopeptidase and chitinase rates, which were used as proxies for microbial activity. Biomass and activity were maximal within the 0- to 3-cm depth range and declined rapidly with sediment depths below 5 cm. Most-probable-number counting using a dilute carbohydrate-containing medium recovered 1.7 to 3.8% of the sediment total bacterial count, with mostly facultatively anaerobic psychrophiles cultured. The median optimal growth temperature for the sediment isolates was 15 degrees C. Many of the isolates identified belonged to genera characteristic of deep-sea habitats, although most appear to be novel species. Phospholipid fatty acid (PLFA) and isoprenoid glycerol dialkyl glycerol tetraether analyses indicated that the samples contained lipid components typical of marine sediments, with profiles varying little between samples at the same depth; however, significant differences in PLFA profiles were found between depths of 0 to 1 cm and 13 to 15 cm, reflecting the presence of a different microbial community. Denaturing gradient gel electrophoresis (DGGE) analysis of amplified bacterial 16S rRNA genes revealed that between samples and across sediment core depths of 1 to 4 cm, the community structure appeared homogenous; however, principal-component analysis of DGGE patterns revealed that at greater sediment depths, successional shifts in community structure were evident. Sequencing of DGGE bands and rRNA probe hybridization analysis revealed that the major community members belonged to delta proteobacteria, putative sulfide oxidizers of the gamma proteobacteria, Flavobacteria, Planctomycetales, and Archaea. rRNA hybridization analyses also indicated that these groups were present at similar levels in the top layer across the shelf region. PMID- 12732511 TI - Biodiversity, community structural shifts, and biogeography of prokaryotes within Antarctic continental shelf sediment. AB - 16S ribosomal DNA (rDNA) clone library analysis was conducted to assess prokaryotic diversity and community structural changes within a surficial sediment core obtained from an Antarctic continental shelf area (depth, 761 m) within the Mertz Glacier Polynya (MGP) region. Libraries were created from three separate horizons of the core (0- to 0.4-cm, 1.5- to 2.5-cm, and 20- to 21-cm depth positions). The results indicated that at the oxic sediment surface (depth, 0 to 0.4 cm) the microbial community appeared to be dominated by a small subset of potentially r-strategist (fast-growing, opportunistic) species, resulting in a lower-than-expected species richness of 442 operational taxonomic units (OTUs). At a depth of 1.5 to 2.5 cm, the species richness (1,128 OTUs) was much higher, with the community dominated by numerous gamma and delta proteobacterial phylotypes. At a depth of 20 to 21 cm, a clear decline in species richness (541 OTUs) occurred, accompanied by a larger number of more phylogenetically divergent phylotypes and a decline in the predominance of Proteobacteria. Based on rRNA and clonal abundance as well as sequence comparisons, syntrophic cycling of oxidized and reduced sulfur compounds appeared to be the dominant process in surficial MGP sediment, as phylotype groups putatively linked to these processes made up a large proportion of clones throughout the core. Between 18 and 65% of 16S rDNA phylotypes detected in a wide range of coastal and open ocean sediments possessed high levels of sequence similarity (>95%) with the MGP sediment phylotypes, indicating that many sediment prokaryote phylotype groups defined in this study are ubiquitous in marine sediment. PMID- 12732512 TI - Virus succession observed during an Emiliania huxleyi bloom. AB - Denaturing gradient gel electrophoresis was used as a molecular tool to determine the diversity and to monitor population dynamics of viruses that infect the globally important coccolithophorid Emiliania huxleyi. We exploited variations in the major capsid protein gene from E. huxleyi-specific viruses to monitor their genetic diversity during an E. huxleyi bloom in a mesocosm experiment off western Norway. We reveal that, despite the presence of several virus genotypes at the start of an E. huxleyi bloom, only a few virus genotypes eventually go on to kill the bloom. PMID- 12732513 TI - Characterization of poly-gamma-glutamate hydrolase encoded by a bacteriophage genome: possible role in phage infection of Bacillus subtilis encapsulated with poly-gamma-glutamate. AB - Some Bacillus subtilis strains, including natto (fermented soybeans) starter strains, produce a capsular polypeptide of glutamate with a gamma-linkage, called poly-gamma-glutamate (gamma-PGA). We identified and purified a monomeric 25-kDa degradation enzyme for gamma-PGA (designated gamma-PGA hydrolase, PghP) from bacteriophage PhiNIT1 in B. subtilis host cells. The monomeric PghP internally hydrolyzed gamma-PGA to oligopeptides, which were then specifically converted to tri-, tetra-, and penta-gamma-glutamates. Monoiodoacetate and EDTA both inhibited the PghP activity, but Zn(2+) or Mn(2+) ions fully restored the enzyme activity inhibited by the chelator, suggesting that a cysteine residue(s) and these metal ions participate in the catalytic mechanism of the enzyme. The corresponding pghP gene was cloned and sequenced from the phage genome. The deduced PghP sequence (208 amino acids) with a calculated M(r) of 22,939 was not significantly similar to any known enzyme. Thus, PghP is a novel gamma-glutamyl hydrolase. Whereas phage PhiNIT1 proliferated in B. subtilis cells encapsulated with gamma-PGA, phage BS5 lacking PghP did not survive well on such cells. Moreover, all nine phages that contaminated natto during fermentation produced PghP, supporting the notion that PghP is important in the infection of natto starters that produce gamma-PGA. Analogous to polysaccharide capsules, gamma-PGA appears to serve as a physical barrier to phage absorption. Phages break down the gamma-PGA barrier via PghP so that phage progenies can easily establish infection in encapsulated cells. PMID- 12732514 TI - Cloning and sequencing of a poly(DL-lactic acid) depolymerase gene from Paenibacillus amylolyticus strain TB-13 and its functional expression in Escherichia coli. AB - The gene encoding a poly(DL-lactic acid) (PLA) depolymerase from Paenibacillus amylolyticus strain TB-13 was cloned and overexpressed in Escherichia coli. The purified recombinant PLA depolymerase, PlaA, exhibited degradation activities toward various biodegradable polyesters, such as poly(butylene succinate), poly(butylene succinate-co-adipate), poly(ethylene succinate), and poly(epsilon caprolactone), as well as PLA. The monomeric lactic acid was detected as the degradation product of PLA. The substrate specificity toward triglycerides and p nitrophenyl esters indicated that PlaA is a type of lipase. The gene encoded 201 amino acid residues, including the conserved pentapeptide Ala-His-Ser-Met-Gly, present in the lipases of mesophilic Bacillus species. The identity of the amino acid sequence of PlaA with Bacillus lipases was no more than 45 to 50%, and some of its properties were different from those of these lipases. PMID- 12732515 TI - Cell culture-Taqman PCR assay for evaluation of Cryptosporidium parvum disinfection. AB - Cryptosporidium parvum represents a challenge to the water industry and a threat to public health. In this study, we developed a cell culture-quantitative PCR assay to evaluate the inactivation of C. parvum with disinfectants. The assay was validated by using a range of disinfectants in common use in the water industry, including low-pressure UV light (LP-UV), ozone, mixed oxidants (MIOX), and chlorine. The assay was demonstrated to be reliable and sensitive, with a lower detection limit of a single infectious oocyst. Effective oocyst inactivation was achieved (>2 log(10) units) with LP-UV (20 mJ/cm(2)) or 2 mg of ozone/liter (for 10 min). MIOX and chlorine treatments of oocysts resulted in minimal effective disinfection, with <0.1 log(10) unit being inactivated. These results demonstrate the inability of MIOX to inactivate Cryptosporidium. The assay is a valuable tool for the evaluation of disinfection systems for drinking water and recycled water. PMID- 12732516 TI - The ftsH gene of the wine bacterium Oenococcus oeni is involved in protection against environmental stress. AB - The wine bacterium Oenococcus oeni has to cope with harsh environmental conditions, including an acidic pH, a high alcoholic content, nonoptimal growth temperatures, and growth-inhibitory compounds such as fatty acids, phenolic acids, and tannins. We describe the characterization and cloning of the O. oeni ftsH gene, encoding a protease belonging to the ATP binding cassette protein superfamily. The O. oeni FtsH protein is closest in sequence similarity to the FtsH homologue of Lactococcus lactis. The O. oeni ftsH gene proved to be stress responsive, since its expression increased at high temperatures or under osmotic shock. O. oeni FtsH protein function was tested in an Escherichia coli ftsH mutant strain, and consistent with the O. oeni ftsH gene expression pattern, the O. oeni FtsH protein provided protection for the E. coli ftsH mutant against heat shock. O. oeni and Bradyrhizobium japonicum FtsH proteins also triggered E. coli resistance to wine toxicity. Genes homologous to O. oeni ftsH were detected in many other lactic acid bacteria found in wine, suggesting that this type of gene constitutes a well-conserved stress-protective molecular device. PMID- 12732517 TI - Global expression profiling and physiological characterization of Corynebacterium glutamicum grown in the presence of L-valine. AB - Addition of L-valine (50 to 200 mM) to glucose minimal medium had no effect on the growth of wild-type Corynebacterium glutamicum ATCC 13032 but inhibited the growth of the derived valine production strain VAL1 [13032 DeltailvA DeltapanBC(pJC1ilvBNCD)] in a concentration-dependent manner. In order to explore this strain-specific valine effect, genomewide expression profiling was performed using DNA microarrays, which showed that valine caused an increased ilvBN mRNA level in VAL1 but not in the wild type. This unexpected result was confirmed by an increased cellular level of the ilvB protein product, i.e., the large subunit of acetohydroxyacid synthase (AHAS), and by an increased AHAS activity of valine treated VAL1 cells. The conclusion that valine caused the limitation of another branched-chain amino acid was confirmed by showing that high concentrations of L isoleucine could relieve the valine effect on VAL1 whereas L-leucine had the same effect as valine. The valine-caused isoleucine limitation was supported by the finding that the inhibitory valine effect was linked to the ilvA deletion that results in isoleucine auxotrophy. Taken together, these results implied that the valine effect is caused by competition for uptake of isoleucine by the carrier BrnQ, which transports all branched-chained amino acids. Indeed, valine inhibition could also be relieved by supplementing VAL1 with the dipeptide isoleucyl-isoleucine, which is taken up by a dipeptide transport system rather than by BrnQ. Interestingly, addition of external valine stimulated valine production by VAL1. This effect is most probably due to a reduced carbon usage for biomass production and to the increased expression of ilvBN, indicating that AHAS activity may still be a limiting factor for valine production in the VAL1 strain. PMID- 12732519 TI - Factors influencing survival of Legionella pneumophila serotype 1 in hot spring water and tap water. AB - The factors involved in the survival of Legionella pneumophila in the microcosms of both hot spring water and tap water were studied by examining cultivability and metabolic activity. L. pneumophila could survive by maintaining metabolic activity but was noncultivable in all microcosms at 42 degrees C, except for one microcosm with a pH of <2.0. Lower temperatures supported survival without loss of cultivability. The cultivability declined with increasing temperature, although metabolic activity was observed at temperatures of up to 45 degrees C. The optimal range of pH for survival was between 6.0 and 8. The metabolic activity could be maintained for long periods even in microcosms with high concentrations of salt. The cultivability of organisms in the post-exponential phase in a tap water microcosm with a low inoculum size was more rapidly reduced than that of organisms in the exponential phase. In contrast, the loss of cultivability in microcosms of a high inoculum size was significant in the exponential phase. Random(ly) amplified polymorphic DNA analysis of microcosms where cultivability was lost but metabolic activity was retained showed no change compared to cells grown freshly, although an effect on the amplified DNA band pattern by production of stress proteins was expected. Resuscitation by the addition of Acanthamoeba castellanii to the microcosm in which cultivability was completely lost but metabolic activity was maintained was observed only in part of the cell population. Our results suggest that L. pneumophila cell populations can potentially survive as free organisms for long periods by maintaining metabolic activity but temporarily losing cultivability under strict environments and requiring resuscitation by ingestion by amoebas. PMID- 12732520 TI - Determining diversity of freshwater fungi on decaying leaves: comparison of traditional and molecular approaches. AB - Traditional microscope-based estimates of species richness of aquatic hyphomycetes depend upon the ability of the species in the community to sporulate. Molecular techniques which detect DNA from all stages of the life cycle could potentially circumvent the problems associated with traditional methods. Leaf disks from red maple, alder, linden, beech, and oak as well as birch wood sticks were submerged in a stream in southeastern Canada for 7, 14, and 28 days. Fungal biomass, estimated by the amount of ergosterol present, increased with time on all substrates. Alder, linden, and maple leaves were colonized earlier and accumulated the highest fungal biomass. Counts and identifications of released conidia suggested that fungal species richness increased, while community evenness decreased, with time (up to 11 species on day 28). Conidia of Articulospora tetracladia dominated. Modifications of two molecular methods-denaturing gradient gel electrophoresis (DGGE) and terminal restriction fragment length polymorphism (T-RFLP) analysis-suggested that both species richness and community evenness decreased with time. The dominant ribotype matched that of A. tetracladia. Species richness estimates based on DGGE were consistently higher than those based on T-RFLP analysis and exceeded those based on spore identification on days 7 and 14. Since traditional and molecular techniques assess different aspects of the fungal organism, both are essential for a balanced view of fungal succession on leaves decaying in streams. PMID- 12732521 TI - Formation of pseudo-terminal restriction fragments, a PCR-related bias affecting terminal restriction fragment length polymorphism analysis of microbial community structure. AB - Terminal restriction fragment length polymorphism (T-RFLP) analysis of PCR amplified genes is a widely used fingerprinting technique in molecular microbial ecology. In this study, we show that besides expected terminal restriction fragments (T-RFs), additional secondary T-RFs occur in T-RFLP analysis of amplicons from cloned 16S rRNA genes at high frequency. A total of 50% of 109 bacterial and 78% of 68 archaeal clones from the guts of cetoniid beetle larvae, using MspI and AluI as restriction enzymes, respectively, were affected by the presence of these additional T-RFs. These peaks were called "pseudo-T-RFs" since they can be detected as terminal fluorescently labeled fragments in T-RFLP analysis but do not represent the primary terminal restriction site as indicated by sequence data analysis. Pseudo-T-RFs were also identified in T-RFLP profiles of pure culture and environmental DNA extracts. Digestion of amplicons with the single-strand-specific mung bean nuclease prior to T-RFLP analysis completely eliminated pseudo-T-RFs. This clearly indicates that single-stranded amplicons are the reason for the formation of pseudo-T-RFs, most probably because single stranded restriction sites cannot be cleaved by restriction enzymes. The strong dependence of pseudo-T-RF formation on the number of cycles used in PCR indicates that (partly) single-stranded amplicons can be formed during amplification of 16S rRNA genes. In a model, we explain how transiently formed secondary structures of single-stranded amplicons may render single-stranded amplicons accessible to restriction enzymes. The occurrence of pseudo-T-RFs has consequences for the interpretation of T-RFLP profiles from environmental samples, since pseudo-T-RFs may lead to an overestimation of microbial diversity. Therefore, it is advisable to establish 16S rRNA gene sequence clone libraries in parallel with T-RFLP analysis from the same sample and to check clones for their in vitro digestion T RF pattern to facilitate the detection of pseudo-T-RFs. PMID- 12732518 TI - Transposon-like organization of the plasmid-borne organophosphate degradation (opd) gene cluster found in Flavobacterium sp. AB - Several bacterial strains that can use organophosphate pesticides as a source of carbon have been isolated from soil samples collected from diverse geographical regions. All these organisms synthesize an enzyme called parathion hydrolase, and in each case the enzyme is encoded by a gene (opd) located on a large indigenous plasmid. These plasmids show considerable genetic diversity, but the region containing the opd gene is highly conserved. Two opd plasmids, pPDL2 from Flavobacterium sp. and pCMS1 from Pseudomonas diminuta, are well characterized, and in each of them a region of about 5.1 kb containing the opd gene shows an identical restriction pattern. We now report the complete sequence of the conserved region of plasmid pPDL2. The opd gene is flanked upstream by an insertion sequence, ISFlsp1, that is a member of the IS21 family, and downstream by a Tn3-like element encoding a transposase and a resolvase. Adjacent to opd but transcribed in the opposite direction is an open reading frame (orf243) with the potential to encode an aromatic hydrolase somewhat similar to Pseudomonas putida TodF. We have shown that orf243 encodes a polypeptide of 27 kDa, which plays a role in the degradation of p-nitrophenol and is likely to act in concert with opd in the degradation of parathion. The linkage of opd and orf243, the organization of the genes flanking opd, and the wide geographical distribution of these genes suggest that this DNA sequence may constitute a complex catabolic transposon. PMID- 12732522 TI - Enhanced killing of Acanthamoeba cysts with a plant peroxidase-hydrogen peroxide halide antimicrobial system. AB - The activity of H(2)O(2) against the resistant cyst stage of the pathogenic free living amoeba Acanthamoeba was enhanced by the addition of KI and either horseradish peroxidase or soybean peroxidase or, to a lesser degree, lactoperoxidase. This resulted in an increase in the cysticidal activity of 3% (wt/vol) H(2)O(2), and there was >3-log killing in 2 h, compared with the 6 h required for comparable results with the peroxide solution alone (P < 0.05). With 2% H(2)O(2), enhancement was observed at all time points (P < 0.05), and total killing of the cyst inoculum occurred at 4 h, compared with 6 h for the peroxide alone. The activity of sublethal 1% H(2)O(2) was enhanced to give 3-log killing after 8 h of exposure (P < 0.05). No enhancement was obtained when KCl or catalase was used as a substitute in the reaction mixtures. The H(2)O(2) was not neutralized in the enhanced system during the experiments. However, in the presence of a platinum disk used to neutralize H(2)O(2) in contact lens care systems, the enhanced 2% H(2)O(2) system gave 2.8-log killing after 6 h or total cyst killing by 8 h, and total neutralization of the H(2)O(2) occurred by 4 h. In contrast, 2% H(2)O(2) alone resulted in <0.8-log killing of cysts in the presence of the platinum disk due to rapid (<1 h) neutralization of the peroxide. Our observations could result in significant improvement in the efficacy of H(2)O(2) contact lens disinfection systems against Acanthamoeba cysts and prevention of acanthamoeba keratitis. PMID- 12732523 TI - Cloning and sequencing of the histidine decarboxylase genes of gram-negative, histamine-producing bacteria and their application in detection and identification of these organisms in fish. AB - The use of molecular tools for early and rapid detection of gram-negative histamine-producing bacteria is important for preventing the accumulation of histamine in fish products. To date, no molecular detection or identification system for gram-negative histamine-producing bacteria has been developed. A molecular method that allows the rapid detection of gram-negative histamine producers by PCR and simultaneous differentiation by single-strand conformation polymorphism (SSCP) analysis using the amplification product of the histidine decarboxylase genes (hdc) was developed. A collection of 37 strains of histamine producing bacteria (8 reference strains from culture collections and 29 isolates from fish) and 470 strains of non-histamine-producing bacteria isolated from fish were tested. Histamine production of bacteria was determined by paper chromatography and confirmed by high-performance liquid chromatography. Among 37 strains of histamine-producing bacteria, all histidine-decarboxylating gram negative bacteria produced a PCR product, except for a strain of Citrobacter braakii. In contrast, none of the non-histamine-producing strains (470 strains) produced an amplification product. Specificity of the amplification was further confirmed by sequencing the 0.7-kbp amplification product. A phylogenetic tree of the isolates constructed using newly determined sequences of partial hdc was similar to the phylogenetic tree generated from 16S ribosomal DNA sequences. Histamine accumulation occurred when PCR amplification of hdc was positive in all of fish samples tested and the presence of powerful histamine producers was confirmed by subsequent SSCP identification. The potential application of the PCR SSCP method as a rapid monitoring tool is discussed. PMID- 12732524 TI - Growth characteristics and intraspecies host specificity of a large virus infecting the dinoflagellate Heterocapsa circularisquama. AB - The growth characteristics and intraspecies host specificity of Heterocapsa circularisquama virus (HcV), a large icosahedral virus specifically infecting the bivalve-killing dinoflagellate H. circularisquama, were examined. Exponentially growing host cells were more sensitive to HcV than those in the stationary phase, and host cells were more susceptible to HcV infection in the culture when a higher percent of the culture was replaced with fresh medium each day, suggesting an intimate relationship between virus sensitivity and the physiological condition of the host cells. HcV was infective over a wide range of temperatures, 15 to 30 degrees C, and the latent period and burst size were estimated at 40 to 56 h and 1,800 to 2,440 infective particles, respectively. Transmission electron microscopy revealed that capsid formation began within 16 h postinfection, and mature virus particles appeared within 24 h postinfection at 20 degrees C. Compared to Heterosigma akashiwo virus, HcV was more widely infectious to H. circularisquama strains that had been independently isolated in the western part of Japan, and only 5.3% of the host-virus combinations (53 host and 10 viral strains) showed resistance to viral infection. The present results are helpful in understanding the ecology of algal host-virus systems in nature. PMID- 12732525 TI - Genetic characterization of Escherichia coli populations from host sources of fecal pollution by using DNA fingerprinting. AB - Escherichia coli isolates were obtained from common host sources of fecal pollution and characterized by using repetitive extragenic palindromic (REP) PCR fingerprinting. The genetic relationship of strains within each host group was assessed as was the relationship of strains among different host groups. Multiple isolates from a single host animal (gull, human, or dog) were found to be identical; however, in some of the animals, additional strains occurred at a lower frequency. REP PCR fingerprint patterns of isolates from sewage (n = 180), gulls (n = 133), and dairy cattle (n = 121) were diverse; within a host group, pairwise comparison similarity indices ranged from 98% to as low as 15%. A composite dendrogram of E. coli fingerprint patterns did not cluster the isolates into distinct host groups but rather produced numerous subclusters (approximately >80% similarity scores calculated with the cosine coefficient) that were nearly exclusive for a host group. Approximately 65% of the isolates analyzed were arranged into host-specific groups. Comparable results were obtained by using enterobacterial repetitive intergenic consensus PCR and pulsed-field gel electrophoresis (PFGE), where PFGE gave a higher differentiation of closely related strains than both PCR techniques. These results demonstrate that environmental studies with genetic comparisons to detect sources of E. coli contamination will require extensive isolation of strains to encompass E. coli strain diversity found in host sources of contamination. These findings will assist in the development of approaches to determine sources of fecal pollution, an effort important for protecting water resources and public health. PMID- 12732526 TI - Insertional mutagenesis of a fungal biocontrol agent led to discovery of a rare cellobiose lipid with antifungal activity. AB - Insertional mutagenesis was applied for the first time to a fungal biocontrol agent, Pseudozyma flocculosa, in an attempt to obtain mutants with altered antagonistic properties. Transformants were obtained via DNA-mediated transformation. Molecular analyses of the transformants revealed that multiple copies of the plasmid were integrated in tandem at one to many chromosomal loci. The transformants were screened for their biocontrol properties using standard bioassays, and the 160 tested transformants were classified into four groups: group I mutants (22 transformants) showed a stronger antagonistic effect than the wild type (WT) while those of group II (107 transformants) had a comparable antagonistic effect; group III mutants (17 transformants) had a decreased antagonistic effect relative to WT and group IV mutants (14 transformants) had lost their biocontrol properties. Culture extracts of the mutants (group IV) and WT were analyzed and compared for the presence of active metabolites which were then separated by solid-phase extraction and purified using conventional methods. Nuclear magnetic resonance experiments and analytical studies on a metabolite specifically produced by the WT revealed the presence of 2-(2',4'-diacetoxy-5' carboxy-pentanoyl) octadecyl cellobioside (flocculosin), a novel glycolipid with strong antifungal properties; the production of this compound would account for the biocontrol activity of P. flocculosa. PMID- 12732527 TI - A novel NAD-dependent dehydrogenase, highly specific for 1,5-anhydro-D-glucitol, from Trichoderma longibrachiatum strain 11-3. AB - A novel NAD-dependent dehydrogenase highly specific for 1,5-anhydro-D-glucitol (1,5-AG) was found in the cell extract of an imperfect fungus, Trichoderma longibrachiatum strain 11-3. This fungus used 1,5-AG as a sole carbon source for growth and transformed 1,5-AG into glucose. 1,5-AG dehydrogenase (AGH) was purified to homogeneity, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The molecular mass of the purified enzyme was estimated to be 36 and 141 kDa by SDS-PAGE and by gel filtration, respectively, suggesting that the enzyme was homotetrameric. The enzyme was highly specific for 1,5-AG and did not exhibit activity with any sugar or sugar alcohol tested in this study other than 1,5-AG. A linear relationship between the initial rate of the enzyme reaction and the concentration of 1,5-AG at the physiological level was observed. The presence of glucose in abundance did not interfere with the relationship. The optimum temperature for the enzyme reaction was 50 degrees C, and the enzyme was stable at temperatures up to 70 degrees C. These results suggested that AGH is a novel enzyme and is useful for specifically diagnosing diabetes mellitus. PMID- 12732528 TI - An exocellular protein from the oil-degrading microbe Acinetobacter venetianus RAG-1 enhances the emulsifying activity of the polymeric bioemulsifier emulsan. AB - The oil-degrading microorganism Acinetobacter venetianus RAG-1 produces an extracellular polyanionic, heteropolysaccharide bioemulsifier termed emulsan. Emulsan forms and stabilizes oil-water emulsions with a variety of hydrophobic substrates. Removal of the protein fraction yields a product, apoemulsan, which exhibits much lower emulsifying activity on hydrophobic substrates such as n hexadecane. One of the key proteins associated with the emulsan complex is a cell surface esterase. The esterase (molecular mass, 34.5 kDa) was cloned and overexpressed in Escherichia coli BL21(DE3) behind the phage T7 promoter with the His tag system. After overexpression, about 80 to 90% of the protein was found in inclusion bodies. The overexpressed esterase was recovered from the inclusion bodies by solubilization with deoxycholate and, after slow dialysis, was purified by metal chelation affinity chromatography. Mixtures containing apoemulsan and either the catalytically active soluble form of the recombinant esterase isolated from cell extracts or the solubilized inactive form of the enzyme recovered from the inclusion bodies formed stable oil-water emulsions with very hydrophobic substrates such as hexadecane under conditions in which emulsan itself was ineffective. Similarly, a series of esterase-defective mutants were generated by site-directed mutagenesis, cloned, and overexpressed in E. coli. Mutant proteins defective in catalytic activity as well as others apparently affected in protein conformation were also active in enhancing the apoemulsan-mediated emulsifying activity. Other proteins, including a His-tagged overexpressed esterase from the related organism Acinetobacter calcoaceticus BD4, showed no enhancement. PMID- 12732529 TI - Naturally occurring bacteria similar to the methyl tert-butyl ether (MTBE) degrading strain PM1 are present in MTBE-contaminated groundwater. AB - Methyl tert-butyl ether (MTBE) is a widespread groundwater contaminant that does not respond well to conventional treatment technologies. Growing evidence indicates that microbial communities indigenous to groundwater can degrade MTBE under aerobic and anaerobic conditions. Although pure cultures of microorganisms able to degrade or cometabolize MTBE have been reported, to date the specific organisms responsible for MTBE degradation in various field studies have not be identified. We report that DNA sequences almost identical (99% homology) to those of strain PM1, originally isolated from a biofilter in southern California, are naturally occurring in an MTBE-polluted aquifer in Vandenberg Air Force Base (VAFB), Lompoc, California. Cell densities of native PM1 (measured by TaqMan quantitative PCR) in VAFB groundwater samples ranged from below the detection limit (in anaerobic sites) to 10(3) to 10(4) cells/ml (in oxygen-amended sites). In groundwater from anaerobic or aerobic sites incubated in microcosms spiked with 10 microg of MTBE/liter, densities of native PM1 increased to approximately 10(5) cells/ml. Native PM1 densities also increased during incubation of VAFB sediments during MTBE degradation. In controlled field plots amended with oxygen, artificially increasing the MTBE concentration was followed by an increase in the in situ native PM1 cell density. This is the first reported relationship between in situ MTBE biodegradation and densities of MTBE-degrading bacteria by quantitative molecular methods. PMID- 12732530 TI - Are readily culturable bacteria in coastal North Sea waters suppressed by selective grazing mortality? AB - We studied the growth of six culturable bacterial lineages from coastal North Sea picoplankton in environmental samples under different incubation conditions. The grazing pressure of heterotrophic nanoflagellates (HNF) was reduced either by double prefiltration through 0.8- micro m-pore-size filters or by 10-fold dilutions with 0.2- micro m (pore-size) prefiltered seawater. We hypothesized that those gamma-proteobacterial genera that are rapidly enriched would also be most strongly affected by HNF regrowth. In the absence of HNF, the mean protein content per bacterial cell increased in both treatments compared to environmental samples, whereas the opposite trend was found in incubations of unaltered seawater. Significant responses to the experimental manipulations were observed in Alteromonas, Pseudoalteromonas, and Vibrio populations. No treatment-specific effects could be detected for members of the Roseobacter group, the Cytophaga latercula-C. marinoflava lineage, or the NOR5 clade. Statistical analysis confirmed a transient increase in the proportions of Alteromonas, Pseudoalteromonas, and Vibrio cells at reduced HNF densities only, followed by an overproportional decline during the phase of HNF regrowth. Cells from these genera were significantly larger than the community average in the dilution treatments, and changes in their relative abundances were negatively correlated with HNF densities. Our findings suggest that bacteria affiliated with frequently isolated genera such as Alteromonas, Pseudoalteromonas, and Vibrio might be rare in coastal North Sea picoplankton because their rapid growth response to changing environmental conditions is counterbalanced by a higher grazing mortality. PMID- 12732531 TI - Automated enumeration of groups of marine picoplankton after fluorescence in situ hybridization. AB - We describe here an automated system for the counting of multiple samples of double-stained microbial cells on sections of membrane filters. The application integrates an epifluorescence microscope equipped with motorized z-axis drive, shutters, and filter wheels with a scanning stage, a digital camera, and image analysis software. The relative abundances of specific microbial taxa are quantified in samples of marine picoplankton, as detected by fluorescence in situ hybridization (FISH) and catalyzed reporter deposition. Pairs of microscopic images are automatically acquired from numerous positions at two wavelengths, and microbial cells with both general DNA and FISH staining are counted after object edge detection and signal-to-background ratio thresholding. Microscopic fields that are inappropriate for cell counting are automatically excluded prior to measurements. Two nested walk paths guide the device across a series of triangular preparations until a user-defined number of total cells has been analyzed per sample. A backup autofocusing routine at incident light allows automated refocusing between individual samples and can reestablish the focal plane after fatal focusing errors at epifluorescence illumination. The system was calibrated to produce relative abundances of FISH-stained cells in North Sea samples that were comparable to results obtained by manual evaluation. Up to 28 preparations could be analyzed within 4 h without operator interference. The device was subsequently applied for the counting of different microbial populations in incubation series of North Sea waters. Automated digital microscopy greatly facilitates the processing of numerous FISH-stained samples and might thus open new perspectives for bacterioplankton population ecology. PMID- 12732533 TI - Use of Bifidobacterium dentium as an indicator of the origin of fecal water pollution. AB - A new, simple, and specific protocol to discriminate between human and animal fecal pollution is described. The procedure is based on the detection of certain Bifidobacterium species in the samples. Two 16S rRNA gene-targeted probes are described. One of these probes (BDE) has as its target a region of the 16S rRNA gene of Bifidobacterium dentium, a Bifidobacterium species of exclusively human origin. The other probe (BAN) is based on the sequence of a region of 16S rRNA gene for several Bifidobacterium species related with animal origins. The specificity of both probes was evaluated by using 24 Bifidobacterium species, and their threshold detection limit was established by DNA-DNA hybridization. DNA-DNA hybridization with the BDE probe showed it to be specific for B. dentium, whereas that with the BAN probe showed it to be specific for B. animalis, B. asteroides, B. coryneforme, B. cuniculi, B. globosum, B. magnum, B. minimum, and B. subtile. A simple and specific protocol was also developed for the detection of their target species in environmental samples (sewage and feces). DNA-DNA hybridization with the BAN probe was only positive for samples from cattle and goats. Thus, this probe is not suitable for the identification of any animal fecal pollution. Whereas all samples with human fecal pollution showed a positive DNA-DNA hybridization result with the BDE probe, none of those with animal fecal pollution did. Therefore, this finding supports the potential use of this probe in detecting fecal pollution of human origin. PMID- 12732532 TI - Genetic organization and molecular analysis of the EcoVIII restriction modification system of Escherichia coli E1585-68 and its comparison with isospecific homologs. AB - The EcoVIII restriction-modification (R-M) system is carried by the Escherichia coli E1585-68 natural plasmid pEC156 (4,312 bp). The two genes were cloned and characterized. The G+C content of the EcoVIII R-M system is 36.1%, which is significantly lower than the average G+C content of either plasmid pEC156 (43.6%) or E. coli genomic DNA (50.8%). The difference suggests that there is a possibility that the EcoVIII R-M system was recently acquired by the genome. The 921-bp EcoVIII endonuclease (R. EcoVIII) gene (ecoVIIIR) encodes a 307-amino-acid protein with an M(r) of 35,554. The convergently oriented EcoVIII methyltransferase (M. EcoVIII) gene (ecoVIIIM) consists of 912 bp that code for a 304-amino-acid protein with an M(r) of 33,930. The exact positions of the start codon AUG were determined by protein microsequencing. Both enzymes recognize the specific palindromic sequence 5'-AAGCTT-3'. Preparations of EcoVIII R-M enzymes purified to homogeneity were characterized. R. EcoVIII acts as a dimer and cleaves a specific sequence between two adenine residues, leaving 4-nucleotide 5' protruding ends. M. EcoVIII functions as a monomer and modifies the first adenine residue at the 5' end of the specific sequence to N(6)-methyladenine. These enzymes are thus functionally identical to the corresponding enzymes of the HindIII (Haemophilus influenzae Rd) and LlaCI (Lactococcus lactis subsp. cremoris W15) R-M systems. This finding is reflected by the levels of homology of M. EcoVIII with M. HindIII and M. LlaCI at the amino acid sequence level (50 and 62%, respectively) and by the presence of nine sequence motifs conserved among m(6) N-adenine beta-class methyltransferases. The deduced amino acid sequence of R. EcoVIII shows weak homology with its two isoschizomers, R. HindIII (26%) and R. LlaCI (17%). A catalytic sequence motif characteristic of restriction endonucleases was found in the primary structure of R. EcoVIII (D(108)X(12)DXK(123)), as well as in the primary structures of R. LlaCI and R. HindIII. Polyclonal antibodies raised against R. EcoVIII did not react with R. HindIII, while anti-M. EcoVIII antibodies cross-reacted with M. LlaCI but not with M. HindIII. R. EcoVIII requires Mg(II) ions for phosphodiester bond cleavage. We found that the same ions are strong inhibitors of the M. EcoVIII enzyme. The biological implications of this finding are discussed. PMID- 12732534 TI - Novel eukaryotic lineages inferred from small-subunit rRNA analyses of oxygen depleted marine environments. AB - Microeukaryotes in oxygen-depleted environments are among the most diverse, as well as the least studied, organisms. We conducted a cultivation-independent, small-subunit (SSU) rRNA-based survey of microeukaryotes in suboxic waters and anoxic sediments in the great Sippewisset salt marsh, Cape Cod, Mass. We generated two clone libraries and analyzed approximately 300 clones, which contained a large diversity of microeukaryotic SSU rRNA signatures. Only a few of these signatures were closely related (sequence similarity of >97%) to the sequences reported earlier. The bulk of our sequences represented deep novel branches within green algae, fungi, cercozoa, stramenopiles, alveolates, euglenozoa and unclassified flagellates. In addition, a significant number of detected rRNA sequences exhibited no affiliation to known organisms and sequences and thus represent novel lineages of the highest taxonomical order, most of them branching off the base of the global phylogenetic tree. This suggests that oxygen depleted environments harbor diverse communities of novel organisms, which may provide an interesting window into the early evolution of eukaryotes. PMID- 12732535 TI - Large-scale spatial distribution of virioplankton in the Adriatic Sea: testing the trophic state control hypothesis. AB - Little is known concerning environmental factors that may control the distribution of virioplankton on large spatial scales. In previous studies workers reported high viral levels in eutrophic systems and suggested that the trophic state is a possible driving force controlling the spatial distribution of viruses. In order to test this hypothesis, we determined the distribution of viral abundance and bacterial abundance and the virus-to-bacterium ratio in a wide area covering the entire Adriatic basin (Mediterranean Sea). To gather additional information on factors controlling viral distribution on a large scale, functional microbial parameters (exoenzymatic activities, bacterial production and turnover) were related to trophic gradients. At large spatial scales, viral distribution was independent of autotrophic biomass and all other environmental parameters. We concluded that in contrast to what was previously hypothesized, changing trophic conditions do not directly affect virioplankton distribution. Since virus distribution was coupled with bacterial turnover times, our results suggest that viral abundance depends on bacterial activity and on host cell abundance. PMID- 12732536 TI - Extent of genetic lesions of the arginine and pyrimidine biosynthetic pathways in Lactobacillus plantarum, L. paraplantarum, L. pentosus, and L. casei: prevalence of CO(2)-dependent auxotrophs and characterization of deficient arg genes in L. plantarum. AB - Lactic acid bacteria require rich media since, due to mutations in their biosynthetic genes, they are unable to synthesize numerous amino acids and nucleobases. Arginine biosynthesis and pyrimidine biosynthesis have a common intermediate, carbamoyl phosphate (CP), whose synthesis requires CO(2). We investigated the extent of genetic lesions in both the arginine biosynthesis and pyrimidine biosynthesis pathways in a collection of lactobacilli, including 150 strains of Lactobacillus plantarum, 32 strains of L. pentosus, 15 strains of L. paraplantarum, and 10 strains of L. casei. The distribution of prototroph and auxotroph phenotypes varied between species. All L. casei strains, no L. paraplantarum strains, two L. pentosus strains, and seven L. plantarum strains required arginine for growth. Arginine auxotrophs were more frequently found in L. plantarum isolated from milk products than in L. plantarum isolated from fermented plant products or humans; association with dairy products might favor arginine auxotrophy. In L. plantarum the argCJBDF genes were functional in most strains, and when they were inactive, only one gene was mutated in more than one half of the arginine auxotrophs. Random mutation may have generated these auxotrophs since different arg genes were inactivated (there were single point mutations in three auxotrophs and nonrevertible genetic lesions in four auxotrophs). These data support the hypothesis that lactic acid bacteria evolve by progressively loosing unnecessary genes upon adaptation to specific habitats, with genome evolution towards cumulative DNA degeneration. Although auxotrophy for only uracil was found in one L. pentosus strain, a high CO(2) requirement (HCR) for arginine and pyrimidine was common; it was found in 74 of 207 Lactobacillus strains tested. These HCR auxotrophs may have had their CP cellular pool-related genes altered or deregulated. PMID- 12732537 TI - A census of rRNA genes and linked genomic sequences within a soil metagenomic library. AB - We have analyzed the diversity of microbial genomes represented in a library of metagenomic DNA from soil. A total of 24,400 bacterial artificial chromosome (BAC) clones were screened for 16S rRNA genes. The sequences obtained from BAC clones were compared with a collection generated by direct PCR amplification and cloning of 16S rRNA genes from the same soil. The results indicated that the BAC library had substantially lower representation of bacteria among the Bacillus, alpha-Proteobacteria, and CFB groups; greater representation among the beta- and gamma-Proteobacteria, and OP10 divisions; and no rRNA genes from the domains Eukaryota and Archaea. In addition to rRNA genes recovered from the bacterial divisions Proteobacteria, Verrucomicrobia, Firmicutes, Cytophagales, and OP11, we identified many rRNA genes from the BAC library affiliated with the bacterial division Acidobacterium; all of these sequences were affiliated with subdivisions that lack cultured representatives. The complete sequence of one BAC clone derived from a member of the Acidobacterium division revealed a complete rRNA operon and 20 other open reading frames, including predicted gene products involved in cell division, cell cycling, folic acid biosynthesis, substrate metabolism, amino acid uptake, DNA repair, and transcriptional regulation. This study is the first step in using genomics to reveal the physiology of as-yet uncultured members of the Acidobacterium division. PMID- 12732539 TI - Production of plant-specific flavanones by Escherichia coli containing an artificial gene cluster. AB - In plants, chalcones are precursors for a large number of flavonoid-derived plant natural products and are converted to flavanones by chalcone isomerase or nonenzymatically. Chalcones are synthesized from tyrosine and phenylalanine via the phenylpropanoid pathway involving phenylalanine ammonia lyase (PAL), cinnamate-4-hydroxylase (C4H), 4-coumarate:coenzyme A ligase (4CL), and chalcone synthase (CHS). For the purpose of production of flavanones in Escherichia coli, three sets of an artificial gene cluster which contained three genes of heterologous origins--PAL from the yeast Rhodotorula rubra, 4CL from the actinomycete Streptomyces coelicolor A3(2), and CHS from the licorice plant Glycyrrhiza echinata--were constructed. The constructions of the three sets were done as follows: (i) PAL, 4CL, and CHS were placed in that order under the control of the T7 promoter (P(T7)) and the ribosome-binding sequence (RBS) in the pET vector, where the initiation codons of 4CL and CHS were overlapped with the termination codons of the preceding genes; (ii) the three genes were transcribed by a single P(T7) in front of PAL, and each of the three contained the RBS at appropriate positions; and (iii) all three genes contained both P(T7) and the RBS. These pathways bypassed C4H, a cytochrome P-450 hydroxylase, because the bacterial 4CL enzyme ligated coenzyme A to both cinnamic acid and 4-coumaric acid. E. coli cells containing the gene clusters produced two flavanones, pinocembrin from phenylalanine and naringenin from tyrosine, in addition to their precursors, cinnamic acid and 4-coumaric acid. Of the three sets, the third gene cluster conferred on the host the highest ability to produce the flavanones. This is a new metabolic engineering technique for the production in bacteria of a variety of compounds of plant and animal origin. PMID- 12732538 TI - Role of Listeria monocytogenes sigma(B) in survival of lethal acidic conditions and in the acquired acid tolerance response. AB - The food-borne pathogen Listeria monocytogenes can acquire enhanced resistance to lethal acid conditions through multiple mechanisms. We investigated contributions of the stress-responsive alternative sigma factor, sigma(B), which is encoded by sigB, to growth phase-dependent acid resistance (AR) and to the adaptive acid tolerance response in L. monocytogenes. At various points throughout growth, we compared the relative survival of L. monocytogenes wild-type and DeltasigB strains that had been exposed to either brain heart infusion (pH 2.5) or synthetic gastric fluid (pH 2.5) with and without prior acid adaptation. Under these conditions, survival of the DeltasigB strain was consistently lower than that of the wild-type strain throughout all phases of growth, ranging from 4 orders of magnitude less in mid-log phase to 2 orders of magnitude less in stationary phase. Survival of both DeltasigB and wild-type L. monocytogenes strains increased by 6 orders of magnitude upon entry into stationary phase, demonstrating that the L. monocytogenes growth phase-dependent AR mechanism is sigma(B) independent. sigma(B)-mediated contributions to acquired acid tolerance appear to be greatest in early logarithmic growth. Loss of a functional sigma(B) reduced the survival of L. monocytogenes at pH 2.5 to a greater extent in the presence of organic acid (100 mM acetic acid) than in the presence of inorganic acid alone (HCl), suggesting that L. monocytogenes protection against organic and inorganic acid may be mediated through different mechanisms. sigma(B) does not appear to contribute to pH(i) homeostasis through regulation of net proton movement across the cell membrane or by regulation of pH(i) buffering by the GAD system under the conditions examined in this study. In summary, a functional sigma(B) protein is necessary for full resistance of L. monocytogenes to lethal acid treatments. PMID- 12732540 TI - Characterization of the 4-hydroxybenzoyl-coenzyme A thioesterase from Arthrobacter sp. strain SU. AB - The Arthrobacter sp. strain SU 4-chlorobenzoate (4-CBA) dehalogenation pathway converts 4-CBA to 4-hydroxybenzoate (4-HBA). The pathway operon contains the genes fcbA, fcbB, and fcbC (A. Schmitz, K. H. Gartemann, J. Fiedler, E. Grund, and R. Eichenlaub, Appl. Environ. Microbiol. 58:4068-4071, 1992). Genes fcbA and fcbB encode 4-CBA-coenzyme A (CoA) ligase and 4-CBA-CoA dehalogenase, respectively, whereas the function of fcbC is not known. We subcloned fcbC and expressed it in Escherichia coli, and we purified and characterized the FcbC protein. A substrate activity screen identified benzoyl-CoA thioesters as the most active substrates. Catalysis of 4-HBA-CoA hydrolysis to 4-HBA and CoA occurred with a k(cat) of 6.7 s(-1) and a K(m) of 1.2 micro M. The k(cat) pH rate profile for 4-HBA-CoA hydrolysis indicated optimal activity over a pH range of 6 to 10. The amino acid sequence of the FcbC protein was compared to other sequences contained in the protein sequence data banks. A large number of sequence homologues of unknown function were identified. On the other hand, the 4 HBA-CoA thioesterases isolated from 4-CBA-degrading Pseudomonas strains did not share significant sequence identity with the FcbC protein, indicating early divergence of the thioesterase-encoding genes. PMID- 12732541 TI - Characterization of Fe(III) reduction by chlororespiring Anaeromyxobacter dehalogenans. AB - Anaeromyxobacter dehalogenans strain 2CP-C has been shown to grow by coupling the oxidation of acetate to the reduction of ortho-substituted halophenols, oxygen, nitrate, nitrite, or fumarate. In this study, strain 2CP-C was also found to grow by coupling Fe(III) reduction to the oxidation of acetate, making it one of the few isolates capable of growth by both metal reduction and chlororespiration. Doubling times for growth of 9.2 and 10.2 h were determined for Fe(III) and 2 chlorophenol reduction, respectively. These were determined by using the rate of [(14)C]acetate uptake into biomass. Fe(III) compounds used by strain 2CP-C include ferric citrate, ferric pyrophosphate, and amorphous ferric oxyhydroxide. The addition of the humic acid analog anthraquinone 2,6-disulfonate (AQDS) increased the reduction rate of amorphous ferric iron oxide, suggesting AQDS was used as an electron shuttle by strain 2CP-C. The addition of chloramphenicol to fumarate-grown cells did not inhibit Fe(III) reduction, indicating that the latter activity is constitutive. In contrast, the addition of chloramphenicol inhibited dechlorination activity, indicating that chlororespiration is inducible. The presence of insoluble Fe(III) oxyhydroxide did not significantly affect dechlorination, whereas the presence of soluble ferric pyrophosphate inhibited dechlorination. With its ability to respire chlorinated organic compounds and metals such as Fe(III), strain 2CP-C is a promising model organism for the study of the interaction of these potentially competing processes in contaminated environments. PMID- 12732542 TI - Electrophoretic analysis of genetic variability within Cryptosporidium parvum from imported and autochthonous cases of human cryptosporidiosis in the United Kingdom. AB - Cryptosporidium parvum oocyst DNA samples (n = 184) from humans with cryptosporidiosis contracted during foreign travel or during outbreaks in the United Kingdom were characterized genetically and categorized by single-strand conformation polymorphism (SSCP)-based analysis of the small-subunit gene (pSSU) (approximately 300 bp) and second internal transcribed spacer (pITS-2) (approximately 230 bp) of nuclear ribosomal DNA. The two recognized genotypes (types 1 and 2) of C. parvum could be readily differentiated by a distinct electrophoretic shift in the pSSU SSCP profile, associated with a nucleotide difference of approximately 1.3 to 1.7%. Of the 102 samples from cases contracted during foreign travel, 88 (86.3%) were identified as C. parvum type 1 and 14 (13.7%) were identified as type 2. For outbreak samples, unequivocal differentiation between type 1 (n = 20; one child nursery outbreak) and type 2 (n = 62; two waterborne outbreaks) was also achieved. Nucleotide variation in pITS-2 (both within and among samples representing each genotype) was substantially greater (10 to 13 different profiles for each genotype, relating to sequence differences of approximately 1 to 42%) than that in pSSU. SSCP analysis of pITS-2 for all samples revealed that some profiles had a broad geographical distribution whereas others were restricted to particular locations, suggesting a link between some subgenotypes and the geographical origin or source. Comparative denaturing polyacrylamide gel electrophoretic analysis revealed the same genotypic identification and a similar subgenotypic classification of samples as SSCP analysis. The findings of this study, particularly the detection of intragenotypic variation by SSCP, should have significant diagnostic implications for investigating transmission patterns and the monitoring of outbreaks. PMID- 12732543 TI - Identification of new genes positively regulated by Tri10 and a regulatory network for trichothecene mycotoxin production. AB - Tri10, a regulatory gene in trichothecene mycotoxin-producing Fusarium species, is required for trichothecene biosynthesis and the coordinated expression of four trichothecene pathway-specific genes (Tri4, Tri5, Tri6, and Tri101) and the isoprenoid biosynthetic gene for farnesyl pyrophosphate synthetase (FPPS). We showed that six more trichothecene genes (Tri3, Tri7, Tri8, Tri9, Tri11, and Tri12) are regulated by Tri10. We also constructed a cDNA library from a strain of Fusarium sporotrichioides that overexpresses Tri10 ( upward arrow Tri10) and used cDNA derived from the upward arrow Tri10 strain and a non-Tri10-expressing strain (DeltaTri10) to differentially screen macroarrays prepared from the cDNA library. This screen identified 15 additional Tri10-regulated transcripts. Four of these transcripts represent Tri1, Tri13, and Tri14 and a gene designated Tri15. Three other sequences are putative orthologs of genes for isoprenoid biosynthesis, the primary metabolic pathway preceding trichothecene biosynthesis. The remaining eight sequences have been designated Ibt (influenced by Tri10) genes. Of the 26 transcripts now known to be positively regulated by Tri10, 22 are positively coregulated by Tri6, a gene that encodes a previously characterized trichothecene pathway-specific transcription factor. These 22 Tri10 and Tri6-coregulated sequences include all of the known Tri genes (except for Tri10), the FPPS gene, and the other three putative isoprenoid biosynthetic genes. Tri6 also regulates a transcript that is not regulated by Tri10. Thus, Tri10 and Tri6 regulate overlapping sets of genes that include a common group of multiple genes for both primary and secondary metabolism. PMID- 12732544 TI - Purification and characterization of streptin, a type A1 lantibiotic produced by Streptococcus pyogenes. AB - Approximately 10% of Streptococcus pyogenes strains inhibit the growth of all nine indicators in a standardized streptococcal bacteriocin typing scheme. The present study has shown that this inhibitory profile, referred to as bacteriocin producer (P)-type 777 activity, is due to the type A1 lantibiotic streptin. Two major forms of streptin were purified to homogeneity from 95% acidified (pH 2) methanol extracts of S. pyogenes M25 cells by using a series of reversed-phase chromatographic separations. The fully processed form of streptin (streptin 1) is a 23-amino-acid peptide with a mass of 2,424 Da. The 2,821-M(r) form of the peptide (streptin 2) has three additional amino acids (TPY) at the N terminus. Strain M25 extracts also contained small quantities of the streptin 1 and streptin 2 peptides in various stages of dehydration. Streptin 1 and streptin 2 were each capable of specifically inducing streptin production when added to strain M25 cultures. The streptin gene cluster resembled that of other type A1 lantibiotics but appeared to lack a streptin-specific proteinase gene. Although the streptin structural gene (srtA) was widespread within S. pyogenes, being detected in 40 of 58 strains, each representing a different M serotype, only 10 of these srtA-positive strains produced active streptin. The failure of some strains to express streptin was attributed to an approximately 4.5-kb deletion in their streptin loci, encompassing genes putatively encoding proteins involved in streptin processing (srtB and srtC) and transport (srtT). In other strains, srtA transcription appeared to be defective. No direct association could be detected between the production of streptin and the production of the lantibiotic-like hemolysin streptolysin S in strain M25. PMID- 12732545 TI - Homologous npdGI genes in 2,4-dinitrophenol- and 4-nitrophenol-degrading Rhodococcus spp. AB - Rhodococcus (opacus) erythropolis HL PM-1 grows on 2,4,6-trinitrophenol or 2,4 dinitrophenol (2,4-DNP) as a sole nitrogen source. The NADPH-dependent F(420) reductase (NDFR; encoded by npdG) and the hydride transferase II (HTII; encoded by npdI) of the strain were previously shown to convert both nitrophenols to their respective hydride Meisenheimer complexes. In the present study, npdG and npdI were amplified from six 2,4-DNP degrading Rhodococcus spp. The genes showed sequence similarities of 86 to 99% to the respective npd genes of strain HL PM-1. Heterologous expression of the npdG and npdI genes showed that they were involved in 2,4-DNP degradation. Sequence analyses of both the NDFRs and the HTIIs revealed conserved domains which may be involved in binding of NADPH or F(420). Phylogenetic analyses of the NDFRs showed that they represent a new group in the family of F(420)-dependent NADPH reductases. Phylogenetic analyses of the HTIIs revealed that they form an additional group in the family of F(420)-dependent glucose-6-phosphate dehydrogenases and F(420)-dependent N(5),N(10) methylenetetrahydromethanopterin reductases. Thus, the NDFRs and the HTIIs may each represent a novel group of F(420)-dependent enzymes involved in catabolism. PMID- 12732546 TI - Genome differences that distinguish Bacillus anthracis from Bacillus cereus and Bacillus thuringiensis. AB - The three species of the group 1 bacilli, Bacillus anthracis, B. cereus, and B. thuringiensis, are genetically very closely related. All inhabit soil habitats but exhibit different phenotypes. B. anthracis is the causative agent of anthrax and is phylogenetically monomorphic, while B. cereus and B. thuringiensis are genetically more diverse. An amplified fragment length polymorphism analysis described here demonstrates genetic diversity among a collection of non-anthrax causing Bacillus species, some of which show significant similarity to B. anthracis. Suppression subtractive hybridization was then used to characterize the genomic differences that distinguish three of the non-anthrax-causing bacilli from B. anthracis Ames. Ninety-three DNA sequences that were present in B. anthracis but absent from the non-anthrax-causing Bacillus genomes were isolated. Furthermore, 28 of these sequences were not found in a collection of 10 non anthrax-causing Bacillus species but were present in all members of a representative collection of B. anthracis strains. These sequences map to distinct loci on the B. anthracis genome and can be assayed simultaneously in multiplex PCR assays for rapid and highly specific DNA-based detection of B. anthracis. PMID- 12732547 TI - Molecular characterization of sulfate-reducing bacteria in the Guaymas Basin. AB - The Guaymas Basin (Gulf of California) is a hydrothermal vent site where thermal alteration of deposited planktonic and terrestrial organic matter forms petroliferous material which supports diverse sulfate-reducing bacteria. We explored the phylogenetic and functional diversity of the sulfate-reducing bacteria by characterizing PCR-amplified dissimilatory sulfite reductase (dsrAB) and 16S rRNA genes from the upper 4 cm of the Guaymas sediment. The dsrAB sequences revealed that there was a major clade closely related to the acetate oxidizing delta-proteobacterial genus Desulfobacter and a clade of novel, deeply branching dsr sequences related to environmental dsr sequences from marine sediments in Aarhus Bay and Kysing Fjord (Denmark). Other dsr clones were affiliated with gram-positive thermophilic sulfate reducers (genus Desulfotomaculum) and the delta-proteobacterial species Desulforhabdus amnigena and Thermodesulforhabdus norvegica. Phylogenetic analysis of 16S rRNAs from the same environmental samples resulted in identification of four clones affiliated with Desulfobacterium niacini, a member of the acetate-oxidizing, nutritionally versatile genus Desulfobacterium, and one clone related to Desulfobacula toluolica and Desulfotignum balticum. Other bacterial 16S rRNA bacterial phylotypes were represented by non-sulfate reducers and uncultured lineages with unknown physiology, like OP9, OP8, as well as a group with no clear affiliation. In summary, analyses of both 16S rRNA and dsrAB clone libraries resulted in identification of members of the Desulfobacteriales in the Guaymas sediments. In addition, the dsrAB sequencing approach revealed a novel group of sulfate reducing prokaryotes that could not be identified by 16S rRNA sequencing. PMID- 12732548 TI - Predictability of Vibrio cholerae in Chesapeake Bay. AB - Vibrio cholerae is autochthonous to natural waters and can pose a health risk when it is consumed via untreated water or contaminated shellfish. The correlation between the occurrence of V. cholerae in Chesapeake Bay and environmental factors was investigated over a 3-year period. Water and plankton samples were collected monthly from five shore sampling sites in northern Chesapeake Bay (January 1998 to February 2000) and from research cruise stations on a north-south transect (summers of 1999 and 2000). Enrichment was used to detect culturable V. cholerae, and 21.1% (n = 427) of the samples were positive. As determined by serology tests, the isolates, did not belong to serogroup O1 or O139 associated with cholera epidemics. A direct fluorescent-antibody assay was used to detect V. cholerae O1, and 23.8% (n = 412) of the samples were positive. V. cholerae was more frequently detected during the warmer months and in northern Chesapeake Bay, where the salinity is lower. Statistical models successfully predicted the presence of V. cholerae as a function of water temperature and salinity. Temperatures above 19 degrees C and salinities between 2 and 14 ppt yielded at least a fourfold increase in the number of detectable V. cholerae. The results suggest that salinity variation in Chesapeake Bay or other parameters associated with Susquehanna River inflow contribute to the variability in the occurrence of V. cholerae and that salinity is a useful indicator. Under scenarios of global climate change, increased climate variability, accompanied by higher stream flow rates and warmer temperatures, could favor conditions that increase the occurrence of V. cholerae in Chesapeake Bay. PMID- 12732549 TI - Bacterial conversion of hydroxylamino aromatic compounds by both lyase and mutase enzymes involves intramolecular transfer of hydroxyl groups. AB - Hydroxylamino aromatic compounds are converted to either the corresponding aminophenols or protocatechuate during the bacterial degradation of nitroaromatic compounds. The origin of the hydroxyl group of the products could be the substrate itself (intramolecular transfer mechanism) or the solvent water (intermolecular transfer mechanism). The conversion of hydroxylaminobenzene to 2 aminophenol catalyzed by a mutase from Pseudomonas pseudoalcaligenes JS45 proceeds by an intramolecular hydroxyl transfer. The conversions of hydroxylaminobenzene to 2- and 4-aminophenol by a mutase from Ralstonia eutropha JMP134 and to 4-hydroxylaminobenzoate to protocatechuate by a lyase from Comamonas acidovorans NBA-10 and Pseudomonas sp. strain 4NT were proposed, but not experimentally proved, to proceed by the intermolecular transfer mechanism. GC-MS analysis of the reaction products formed in H(2)(18)O did not indicate any (18)O-label incorporation during the conversion of hydroxylaminobenzene to 2- and 4-aminophenols catalyzed by the mutase from R. eutropha JMP134. During the conversion of 4-hydroxylaminobenzoate catalyzed by the hydroxylaminolyase from Pseudomonas sp. strain 4NT, only one of the two hydroxyl groups in the product, protocatechuate, was (18)O labeled. The other hydroxyl group in the product must have come from the substrate. The mutase in strain JS45 converted 4 hydroxylaminobenzoate to 4-amino-3-hydroxybenzoate, and the lyase in Pseudomonas strain 4NT converted hydroxylaminobenzene to aniline and 2-aminophenol but not to catechol. The results indicate that all three types of enzyme-catalyzed rearrangements of hydroxylamino aromatic compounds proceed via intramolecular transfer of hydroxyl groups. PMID- 12732550 TI - Origin of contamination and genetic diversity of Escherichia coli in beef cattle. AB - The possible origin of beef contamination and genetic diversity of Escherichia coli populations in beef cattle, on carcasses and ground beef, was examined by using random amplification of polymorphic DNA (RAPD) and PCR-restriction fragment length polymorphism (PCR-RFLP) analysis of the fliC gene. E. coli was recovered from the feces of 10 beef cattle during pasture grazing and feedlot finishing and from hides, carcasses, and ground beef after slaughter. The 1,403 E. coli isolates (855 fecal, 320 hide, 153 carcass, and 75 ground beef) were grouped into 121 genetic subtypes by using the RAPD method. Some of the genetic subtypes in cattle feces were also recovered from hides, prechilled carcasses, chilled carcasses, and ground beef. E. coli genetic subtypes were shared among cattle at all sample times, but a number of transient types were unique to individual animals. The genetic diversity of the E. coli population changed over time within individual animals grazing on pasture and in the feedlot. Isolates from one animal (59 fecal, 30 hide, 19 carcass, and 12 ground beef) were characterized by the PCR-RFLP analysis of the fliC gene and were grouped into eight genotypes. There was good agreement between the results obtained with the RAPD and PCR-RFLP techniques. In conclusion, the E. coli contaminating meat can originate from cattle feces, and the E. coli population in beef cattle was highly diverse. Also, genetic subtypes can be shared among animals or can be unique to an animal, and they are constantly changing. PMID- 12732552 TI - Purification and characterization of an inverting stereo- and enantioselective sec-alkylsulfatase from the gram-positive bacterium Rhodococcus ruber DSM 44541. AB - Whole cells of Rhodococcus ruber DSM 44541 were found to hydrolyze (+/-)-2-octyl sulfate in a stereo- and enantiospecific fashion. When growing on a complex medium, the cells produced two sec-alkylsulfatases and (at least) one prim alkylsulfatase in the absence of an inducer, such as a sec-alkyl sulfate or a sec alcohol. From the crude cell-free lysate, two proteins responsible for sulfate ester hydrolysis (designated RS1 and RS2) were separated from each other based on their different hydrophobicities and were subjected to further chromatographic purification. In contrast to sulfatase RS1, enzyme RS2 proved to be reasonably stable and thus could be purified to homogeneity. Sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed a single band at a molecular mass of 43 kDa. Maximal enzyme activity was observed at 30 degrees C and at pH 7.5. Sulfatase RS2 showed a clear preference for the hydrolysis of linear secondary alkyl sulfates, such as 2-, 3-, or 4-octyl sulfate, with remarkable enantioselectivity (an enantiomeric ratio of up to 21 [23]). Enzymatic hydrolysis of (R)-2-octyl sulfate furnished (S)-2-octanol without racemization, which revealed that the enzymatic hydrolysis proceeded through inversion of the configuration at the stereogenic carbon atom. Screening of a broad palette of potential substrates showed that the enzyme exhibited limited substrate tolerance; while simple linear sec-alkyl sulfates (C(7) to C(10)) were freely accepted, no activity was found with branched and mixed aryl-alkyl sec-sulfates. Due to the fact that prim-sulfates were not accepted, the enzyme was classified as sec-alkylsulfatase (EC 3.1.6.X). PMID- 12732553 TI - Impact of land use intensity on the species diversity of arbuscular mycorrhizal fungi in agroecosystems of Central Europe. AB - The impact of land use intensity on the diversity of arbuscular mycorrhizal fungi (AMF) was investigated at eight sites in the "three-country corner" of France, Germany, and Switzerland. Three sites were low-input, species-rich grasslands. Two sites represented low- to moderate-input farming with a 7-year crop rotation, and three sites represented high-input continuous maize monocropping. Representative soil samples were taken, and the AMF spores present were morphologically identified and counted. The same soil samples also served as inocula for "AMF trap cultures" with Plantago lanceolata, Trifolium pratense, and Lolium perenne. These trap cultures were established in pots in a greenhouse, and AMF root colonization and spore formation were monitored over 8 months. For the field samples, the numbers of AMF spores and species were highest in the grasslands, lower in the low- and moderate-input arable lands, and lowest in the lands with intensive continuous maize monocropping. Some AMF species occurred at all sites ("generalists"); most of them were prevalent in the intensively managed arable lands. Many other species, particularly those forming sporocarps, appeared to be specialists for grasslands. Only a few species were specialized on the arable lands with crop rotation, and only one species was restricted to the high input maize sites. In the trap culture experiment, the rate of root colonization by AMF was highest with inocula from the permanent grasslands and lowest with those from the high-input monocropping sites. In contrast, AMF spore formation was slowest with the former inocula and fastest with the latter inocula. In conclusion, the increased land use intensity was correlated with a decrease in AMF species richness and with a preferential selection of species that colonized roots slowly but formed spores rapidly. PMID- 12732551 TI - The ars detoxification system is advantageous but not required for As(V) respiration by the genetically tractable Shewanella species strain ANA-3. AB - Arsenate [As(V); HAsO(4)(2-)] respiration by bacteria is poorly understood at the molecular level largely due to a paucity of genetically tractable organisms with this metabolic capability. We report here the isolation of a new As(V)-respiring strain (ANA-3) that is phylogenetically related to members of the genus Shewanella and that also provides a useful model system with which to explore the molecular basis of As(V) respiration. This gram-negative strain stoichiometrically couples the oxidation of lactate to acetate with the reduction of As(V) to arsenite [As(III); HAsO(2)]. The generation time and lactate molar growth yield (Y(lactate)) are 2.8 h and 10.0 g of cells mol of lactate(-1), respectively, when it is grown anaerobically on lactate and As(V). ANA-3 uses a wide variety of terminal electron acceptors, including oxygen, soluble ferric iron, oxides of iron and manganese, nitrate, fumarate, the humic acid functional analog 2,6-anthraquinone disulfonate, and thiosulfate. ANA-3 also reduces As(V) to As(III) in the presence of oxygen and resists high concentrations of As(III) (up to 10 mM) when grown under either aerobic or anaerobic conditions. ANA-3 possesses an ars operon (arsDABC) that allows it to resist high levels of As(III); this operon also confers resistance to the As-sensitive strains Shewanella oneidensis MR-1 and Escherichia coli AW3110. When the gene encoding the As(III) efflux pump, arsB, is inactivated in ANA-3 by a polar mutation that also eliminates the expression of arsC, which encodes an As(V) reductase, the resulting As(III)-sensitive strain still respires As(V); however, the generation time and the Y(lactate) value are two- and threefold lower, respectively, than those of the wild type. These results suggest that ArsB and ArsC may be useful for As(V)-respiring bacteria in environments where As concentrations are high, but that neither is required for respiration. PMID- 12732556 TI - Recovery and enumeration of Cryptosporidium parvum from animal fecal matrices. AB - Accurate quantification of Cryptosporidium parvum oocysts in animal fecal deposits on land is an essential starting point for estimating watershed C. parvum loads. Due to the general poor performance and variable recovery efficiency of existing enumeration methods, protocols were devised based on initial dispersion of oocysts from feces by vortexing in 2 mM tetrasodium pyrophosphate, followed by immunomagnetic separation. The protocols were validated by using an internal control seed preparation to determine the levels of oocyst recovery for a range of fecal types. The levels of recovery of 10(2) oocysts from cattle feces (0.5 g of processed feces) ranged from 31 to 46%, and the levels of recovery from sheep feces (0.25 g of processed feces) ranged from 21% to 35%. The within-sample coefficients of variation for the percentages of recovery from five replicates ranged from 10 to 50%. The ranges for levels of recovery of oocysts from cattle, kangaroo, pig, and sheep feces (juveniles and adults) collected in a subsequent watershed animal fecal survey were far wider than the ranges predicted by the validation data. Based on the use of an internal control added to each fecal sample, the levels of recovery ranged from 0 to 83% for cattle, from 4 to 62% for sheep, from 1 to 42% for pigs, and from 40 to 73% for kangaroos. Given the variation in the levels of recovery of oocysts from different fecal matrices, it is recommended that an internal control be added to at least one replicate of every fecal sample analyzed to determine the percentage of recovery. Depending on the animal type and based on the lowest approximate percentages of recovery, between 10 and 100 oocysts g of feces(-1) must be present to be detected. PMID- 12732555 TI - Expression of a cloned cyclopropane fatty acid synthase gene reduces solvent formation in Clostridium acetobutylicum ATCC 824. AB - The cyclopropane fatty acid synthase gene (cfa) of Clostridium acetobutylicum ATCC 824 was cloned and overexpressed under the control of the clostridial ptb promoter. The function of the cfa gene was confirmed by complementation of an Escherichia coli cfa-deficient strain in terms of fatty acid composition and growth rate under solvent stress. Constructs expressing cfa were introduced into C. acetobutylicum hosts and cultured in rich glucose broth in static flasks without pH control. Overexpression of the cfa gene in the wild type and in a butyrate kinase-deficient strain increased the cyclopropane fatty acid content of early-log-phase cells as well as initial acid and butanol resistance. However, solvent production in the cfa-overexpressing strain was considerably decreased, while acetate and butyrate levels remained high. The findings suggest that overexpression of cfa results in changes in membrane properties that dampen the full induction of solventogenesis. The overexpression of a marR homologous gene preceding the cfa gene in the clostridial genome resulted in reduced cyclopropane fatty acid accumulation. PMID- 12732554 TI - Association of Borrelia garinii and B. valaisiana with songbirds in Slovakia. AB - In Europe, 6 of the 11 genospecies of Borrelia burgdorferi sensu lato are prevalent in questing Ixodes ricinus ticks. In most parts of Central Europe, B. afzelii, B. garinii, and B. valaisiana are the most frequent species, whereas B. burgdorferi sensu stricto, B. bissettii, and B. lusitaniae are rare. Previously, it has been shown that B. afzelii is associated with European rodents. Therefore, the aim of this study was to identify reservoir hosts of B. garinii and B. valaisiana in Slovakia. Songbirds were captured in a woodland near Bratislava and investigated for engorged ticks. Questing I. ricinus ticks were collected in the same region. Both tick pools were analyzed for spirochete infections by PCR, followed by DNA-DNA hybridization and, for a subsample, by nucleotide sequencing. Three of the 17 captured songbird species were infested with spirochete-infected ticks. Spirochetes in ticks that had fed on birds were genotyped as B. garinii and B. valaisiana, whereas questing ticks were infected with B. afzelii, B. garinii, and B. valaisiana. Furthermore, identical ospA alleles of B. garinii were found in ticks that had fed on the birds and in questing ticks. The data show that songbirds are reservoir hosts of B. garinii and B. valaisiana but not of B. afzelii. This and previous studies confirm that B. burgdorferi sensu lato is host associated and that this bacterial species complex contains different ecotypes. PMID- 12732557 TI - Optimization of single-base-pair mismatch discrimination in oligonucleotide microarrays. AB - The discrimination between perfect-match and single-base-pair-mismatched nucleic acid duplexes was investigated by using oligonucleotide DNA microarrays and nonequilibrium dissociation rates (melting profiles). DNA and RNA versions of two synthetic targets corresponding to the 16S rRNA sequences of Staphylococcus epidermidis (38 nucleotides) and Nitrosomonas eutropha (39 nucleotides) were hybridized to perfect-match probes (18-mer and 19-mer) and to a set of probes having all possible single-base-pair mismatches. The melting profiles of all probe-target duplexes were determined in parallel by using an imposed temperature step gradient. We derived an optimum wash temperature for each probe and target by using a simple formula to calculate a discrimination index for each temperature of the step gradient. This optimum corresponded to the output of an independent analysis using a customized neural network program. These results together provide an experimental and analytical framework for optimizing mismatch discrimination among all probes on a DNA microarray. PMID- 12732558 TI - A flow-cytometric gram-staining technique for milk-associated bacteria. AB - A Gram-staining technique combining staining with two fluorescent stains, Oregon Green-conjugated wheat germ agglutinin (WGA) and hexidium iodide (HI) followed by flow-cytometric detection is described. WGA stains gram-positive bacteria while HI binds to the DNA of all bacteria after permeabilization by EDTA and incubation at 50 degrees C for 15 min. For WGA to bind to gram-positive bacteria, a 3 M potassium chloride solution was found to give the highest fluorescence intensity. A total of 12 strains representing some of the predominant bacterial species in bulk tank milk and mixtures of these were stained and analyzed by flow cytometry. Overall, the staining method showed a clear differentiation between gram-positive and gram-negative bacterial populations. For stationary-stage cultures of seven gram-positive bacteria and five gram-negative bacteria, an average of 99% of the cells were correctly interpreted. The method was only slightly influenced by the growth phase of the bacteria or conditions such as freezing at -18 degrees C for 24 h. For any of these conditions, an average of at least 95% of the cells were correctly interpreted. When stationary-stage cultures were stored at 5 degrees C for 14 days, an average of 86% of the cells were correctly interpreted. The Gram staining technique was applied to the flow cytometry analysis of bulk tank milk inoculated with Staphylococcus aureus and Escherichia coli. These results demonstrate that the technique is suitable for analyzing milk samples without precultivation. PMID- 12732560 TI - Protocol for rapid fluorescence in situ hybridization of bacteria in cryosections of microarthropods. AB - A protocol was developed to detect bacteria inhabiting microarthropods by means of small-subunit rRNA-targeted fluorescence in situ hybridization and microscopy. The protocol is based on cryosections of whole specimens. In contrast to more commonly applied paraffin-embedding techniques, the protocol is quicker and reduces the number of manipulations which might damage the microscopic material. The method allowed the study of the bacterial colonization of Folsomia candida (Collembola) and the detection of bacteria in both the gut and tissue. PMID- 12732559 TI - In vivo tracking of Campylobacter jejuni by using a novel recombinant expressing green fluorescent protein. AB - Campylobacter jejuni is a leading cause of food-borne disease in developed countries. The goal of this study was to develop a plasmid-based reporter system with green fluorescent protein (GFP) to facilitate the study of C. jejuni in a variety of niches. C. jejuni transformants harboring the pMEK91 GFP gene (gfp) containing vector were readily detectable by both fluorescence microscopy and flow cytometry. Given the ease of detecting these organisms, additional experiments were performed in which BALB/c mice were injected intraperitoneally with C. jejuni harboring the gfp-containing vector. Four hours after injection of the mice, flow cytometry analyses determined that C. jejuni synthesizing GFP were predominantly associated with granulocytes. More specifically, the proportion of CD11b(+) Gr-1(+) lavage neutrophils with green fluorescence ranged from 99.7 to 100%, while the proportion of CD11b(+) Gr-1(-) lavage macrophages ranged from 77.0 to 80.0%. In contrast, few CD11b(-) CD45R(+) B lymphocytes from the lavage of the C. jejuni-injected mice were associated with green-fluorescent C. jejuni (proportions ranged from 0.75 to 0.77%). Cell-free C. jejuni was recovered from tissue homogenates after intraperitoneal injection. Macrorestriction profiling with pulsed-field gel electrophoresis identified a genotypic variant of the C. jejuni F38011 wild-type isolate. In vivo this variant displayed a phenotype identical to that of the wild-type isolate. In summary, we demonstrate that C. jejuni associates with marker-defined cellular subsets in vivo with a novel gfp reporter system and that C. jejuni genotypic variants can be isolated from both in vitro and in vivo systems. PMID- 12732562 TI - Recombinant Saccharomyces cerevisiae expressing P450 in artificial digestive systems: a model for biodetoxication in the human digestive environment. AB - The use of genetically engineered microorganisms such as bacteria or yeasts as live vehicles to carry out bioconversion directly in the digestive environment is an important challenge for the development of innovative biodrugs. A system that mimics the human gastrointestinal tract was combined with a computer simulation to evaluate the survival rate and cinnamate 4-hydroxylase activity of a recombinant model of Saccharomyces cerevisiae expressing the plant P450 73A1. The yeasts showed a high level of resistance to gastric and small intestinal secretions (survival rate after 4 h of digestion, 95.6% +/- 10.1% [n = 4]) but were more sensitive to the colonic conditions (survival rate after 4 h of incubation, 35.9% +/- 2.7% [n = 3]). For the first time, the ability of recombinant S. cerevisiae to carry out a bioconversion reaction has been demonstrated throughout the gastrointestinal tract. In the gastric-small intestinal system, 41.0% +/- 5.8% (n = 3) of the ingested trans-cinnamic acid was converted into p-coumaric acid after 4 h of digestion, as well as 8.9% +/- 1.6% (n = 3) in the stomach, 13.8% +/- 3.3% (n = 3) in the duodenum, 11.8% +/- 3.4% (n = 3) in the jejunum, and 6.5% +/- 1.0% (n = 3) in the ileum. In the large intestinal system, cinnamate 4-hydroxylase activity was detected but was too weak to be quantified. These results suggest that S. cerevisiae may afford a useful host for the development of biodrugs and may provide an innovative system for the prevention or treatment of diseases that escape classical drug action. In particular, yeasts may provide a suitable vector for biodetoxication in the digestive environment. PMID- 12732561 TI - Specific detection of Dehalococcoides species by fluorescence in situ hybridization with 16S rRNA-targeted oligonucleotide probes. AB - Dehalococcoides ethenogenes is the only known cultivated organism capable of complete dehalogenation of tetrachloroethene (PCE) to ethene. The prevalence of Dehalococcoides species in the environment and their association with complete dehalogenation of chloroethenes suggest that they play an important role in natural attenuation of chloroethenes and are promising candidates for engineered bioremediation of these contaminants. Both natural attenuation and bioremediation require reliable and sensitive methods to monitor the presence, distribution, and fate of the organisms of interest. Here we report the development of 16S rRNA targeted oligonucleotide probes for Dehalococcoides species. The two designed probes together encompass 28 sequences of 16S rRNA genes retrieved from the public database. Except D. ethenogenes and CBDB1, all the others are environmental clones obtained from sites contaminated with chlorinated ethenes. They are all closely related and form a unique cluster of Dehalococcoides species. In situ hybridization of probe Dhe1259t with D. ethenogenes strain 195 and two enrichment cultures demonstrated the applicability of the probe to monitoring the abundance of active Dehalococcoides species in these enrichment samples. PMID- 12732563 TI - Cyanobacterial ecotypes in different optical microenvironments of a 68 degrees C hot spring mat community revealed by 16S-23S rRNA internal transcribed spacer region variation. AB - We examined the population of unicellular cyanobacteria (Synechococcus) in the upper 3-mm vertical interval of a 68 degrees C region of a microbial mat in a hot spring effluent channel (Yellowstone National Park, Wyoming). Fluorescence microscopy and microsensor measurements of O(2) and oxygenic photosynthesis demonstrated the existence of physiologically distinct Synechococcus populations at different depths along a light gradient quantified by scalar irradiance microprobes. Molecular methods were used to evaluate whether physiologically distinct populations could be correlated with genetically distinct populations over the vertical interval. We were unable to identify patterns in genetic variation in Synechococcus 16S rRNA sequences that correlate with different vertically distributed populations. However, patterns of variation at the internal transcribed spacer locus separating 16S and 23S rRNA genes suggested the existence of closely related but genetically distinct populations corresponding to different functional populations occurring at different depths. PMID- 12732564 TI - In situ accessibility of Saccharomyces cerevisiae 26S rRNA to Cy3-labeled oligonucleotide probes comprising the D1 and D2 domains. AB - Fluorescence in situ hybridization (FISH) has proven to be most useful for the identification of microorganisms. However, species-specific oligonucleotide probes often fail to give satisfactory results. Among the causes leading to low hybridization signals is the reduced accessibility of the targeted rRNA site to the oligonucleotide, mainly for structural reasons. In this study we used flow cytometry to determine whole-cell fluorescence intensities with a set of 32 Cy3 labeled oligonucleotide probes covering the full length of the D1 and D2 domains in the 26S rRNA of Saccharomyces cerevisiae PYCC 4455(T). The brightest signal was obtained with a probe complementary to positions 223 to 240. Almost half of the probes conferred a fluorescence intensity above 60% of the maximum, whereas only one probe could hardly detect the cells. The accessibility map based on the results obtained can be extrapolated to other yeasts, as shown experimentally with 27 additional species (14 ascomycetes and 13 basidiomycetes). This work contributes to a more rational design of species-specific probes for yeast identification and monitoring. PMID- 12732565 TI - Isolation and characterization of novel psychrophilic, neutrophilic, Fe oxidizing, chemolithoautotrophic alpha- and gamma-proteobacteria from the deep sea. AB - We report the isolation and physiological characterization of novel, psychrophilic, iron-oxidizing bacteria (FeOB) from low-temperature weathering habitats in the vicinity of the Juan de Fuca deep-sea hydrothermal area. The FeOB were cultured from the surfaces of weathered rock and metalliferous sediments. They are capable of growth on a variety of natural and synthetic solid rock and mineral substrates, such as pyrite (FeS(2)), basalt glass ( approximately 10 wt% FeO), and siderite (FeCO(3)), as their sole energy source, as well as numerous aqueous Fe substrates. Growth temperature characteristics correspond to the in situ environmental conditions of sample origin; the FeOB grow optimally at 3 to 10 degrees C and at generation times ranging from 57 to 74 h. They are obligate chemolithoautotrophs and grow optimally under microaerobic conditions in the presence of an oxygen gradient or anaerobically in the presence of nitrate. None of the strains are capable of using any organic or alternate inorganic substrates tested. The bacteria are phylogenetically diverse and have no close Fe-oxidizing or autotrophic relatives represented in pure culture. One group of isolates are gamma-Proteobacteria most closely related to the heterotrophic bacterium Marinobacter aquaeolei (87 to 94% sequence similarity). A second group of isolates are alpha-Proteobacteria most closely related to the deep-sea heterotrophic bacterium Hyphomonas jannaschiana (81 to 89% sequence similarity). This study provides further evidence for the evolutionarily widespread capacity for Fe oxidation among bacteria and suggests that FeOB may play an unrecognized geomicrobiological role in rock weathering in the deep sea. PMID- 12732566 TI - Development of a plating medium for selection of Helicobacter pylori from water samples. AB - The goal of this study was to develop a simple plating medium to allow large scale screening of water samples for the presence of Helicobacter pylori. Five conventional plating media (brain heart infusion, brucella agar, Columbia blood agar base, campylobacter agar kit Skirrow, and HPSPA medium), each containing a commercial antibiotic supplement, were initially evaluated. Eight strains selected as common waterborne organisms (Acinetobacter, Aeromonas, Bacillus, Escherichia coli, Enterobacter, Enterococcus, Helicobacter pylori, and Pseudomonas strains) were individually plated onto each of these media. Three organisms (Acinetobacter, E. coli, and H. pylori) were able to grow on all five media. This growth was unacceptable since Helicobacter grows very slowly and competing organisms must be inhibited for up to 7 days. Therefore, a more selective medium (HP agar) containing a novel mixture of growth supplements plus amphotericin B and polymyxin B was developed. This medium also included a phenol red color indicator for urease production. Aliquots of nonsterile well water that contained native flora (Flavobacterium, Serratia, Citrobacter, Pasteurella, Ochrobactrum, Rahnella, and unidentified molds) and were further adulterated with the eight strains listed above (10(6) CFU of each strain per 100 ml) were spiked with H. pylori and were plated. In spite of the heavy mixed microbial load, only H. pylori colonies grew during 7 days of incubation at 37 degrees C. The color indicator system allowed presumptive identification of H. pylori colonies sooner (12 to 20 h) than the conventional media tested allowed. The HP formulation developed in this study provides a medium with superior selectivity for H. pylori from mixed microbial populations in water and reduces the time required to complete the assay. PMID- 12732567 TI - Environmental surveillance system to track wild poliovirus transmission. AB - Eradication of poliomyelitis from large metropolis cities in India has been difficult due to high population density and the presence of large urban slums. Three paralytic poliomyelitis cases were reported in Mumbai, India, in 1999 and 2000 in spite of high immunization coverage and good-quality supplementary immunization activities. We therefore established a systematic environmental surveillance study by weekly screening of sewage samples from three high-risk slum areas to detect the silent transmission of wild poliovirus. In 2001, from among the 137 sewage samples tested, wild poliovirus type 1 was isolated from 35 and wild poliovirus type 3 was isolated from 1. Acute flaccid paralysis (AFP) surveillance indicated one case of paralytic poliomyelitis from the city. Phylogenetic analysis with complete VP1 sequences revealed that the isolates from environmental samples belonged to four lineages of wild polioviruses recently isolated from poliomyelitis cases in Uttar Pradesh and not to those previously isolated from AFP cases in Mumbai. Wild poliovirus thus introduced caused one case of paralytic poliomyelitis. The virus was detected in environmental samples 3 months before. It was found that wild polioviruses introduced several times during the year circulated in Mumbai for a limited period before being eliminated. Environmental surveillance was found to be sensitive for the detection of wild poliovirus silent transmission. Nucleotide sequence analysis helped identify wild poliovirus reservoir areas. PMID- 12732568 TI - An improved protocol for quantification of freshwater Actinobacteria by fluorescence in situ hybridization. AB - We tested a previously described protocol for fluorescence in situ hybridization of marine bacterioplankton with horseradish peroxidase-labeled rRNA-targeted oligonucleotide probes and catalyzed reporter deposition (CARD-FISH) in plankton samples from different lakes. The fraction of Bacteria detected by CARD-FISH was significantly lower than after FISH with fluorescently monolabeled probes. In particular, the abundances of aquatic Actinobacteria were significantly underestimated. We thus developed a combined fixation and permeabilization protocol for CARD-FISH of freshwater samples. Enzymatic pretreatment of fixed cells was optimized for the controlled digestion of gram-positive cell walls without causing overall cell loss. Incubations with high concentrations of lysozyme (10 mg ml(-1)) followed by achromopeptidase (60 U ml(-1)) successfully permeabilized cell walls of Actinobacteria for subsequent CARD-FISH both in enrichment cultures and environmental samples. Between 72 and >99% (mean, 86%) of all Bacteria could be visualized with the improved assay in surface waters of four lakes. For freshwater samples, our method is thus superior to the CARD-FISH protocol for marine Bacteria (mean, 55%) and to FISH with directly fluorochrome labeled probes (mean, 67%). Actinobacterial abundances in the studied systems, as detected by the optimized protocol, ranged from 32 to >55% (mean, 45%). Our findings confirm that members of this lineage are among the numerically most important Bacteria of freshwater picoplankton. PMID- 12732569 TI - Identification of an emergent and atypical Pseudomonas viridiflava lineage causing bacteriosis in plants of agronomic importance in a Spanish region. AB - Pseudomonas strains with an atypical LOPAT profile (where LOPAT is a series of determinative tests: L, levan production; O, oxidase production; P, pectinolitic activity; A, arginine dihydrolase production; and T, tobacco hypersensibility) can be regarded as emergent pathogens in the Principality of Asturias (Spain), where they have been causing, since 1999, severe damage in at least three taxonomically unrelated orchard plants of agronomic importance: common bean (Phaseolus vulgaris), kiwifruit (Actinidia deliciosa), and lettuce (Lactuca sativa). These strains are mainly differentiated by production of yellowish mucoid material in hypersucrose medium, used for the levan test, and by a variable pectinolytic activity on different potato varieties. The atypical organisms were identified as Pseudomonas viridiflava based on their 16S rRNA sequences. Among them a certain intraspecies genetic heterogeneity was detected by randomly amplified polymorphic DNA (RAPD) typing. To differentiate between isolates of P. viridiflava and Pseudomonas syringae pathovars, a 16S ribosomal DNA restriction fragment length polymorphism method employing the restriction endonucleases SacI and HinfI was developed. This could be used as a means of reliable species determination after the usual phenotypical characterization, which includes the LOPAT tests. PMID- 12732570 TI - Stable carbon isotope fractionation by sulfate-reducing bacteria. AB - Biogeochemical transformations occurring in the anoxic zones of stratified sedimentary microbial communities can profoundly influence the isotopic and organic signatures preserved in the fossil record. Accordingly, we have determined carbon isotope discrimination that is associated with both heterotrophic and lithotrophic growth of pure cultures of sulfate-reducing bacteria (SRB). For heterotrophic-growth experiments, substrate consumption was monitored to completion. Sealed vessels containing SRB cultures were harvested at different time intervals, and delta(13)C values were determined for gaseous CO(2), organic substrates, and products such as biomass. For three of the four SRB, carbon isotope effects between the substrates, acetate or lactate and CO(2), and the cell biomass were small, ranging from 0 to 2 per thousand. However, for Desulfotomaculum acetoxidans, the carbon incorporated into biomass was isotopically heavier than the available substrates by 8 to 9 per thousand. SRB grown lithoautotrophically consumed less than 3% of the available CO(2) and exhibited substantial discrimination (calculated as isotope fractionation factors [alpha]), as follows: for Desulfobacterium autotrophicum, alpha values ranged from 1.0100 to 1.0123; for Desulfobacter hydrogenophilus, the alpha value was 0.0138, and for Desulfotomaculum acetoxidans, the alpha value was 1.0310. Mixotrophic growth of Desulfovibrio desulfuricans on acetate and CO(2) resulted in biomass with a delta(13)C composition intermediate to that of the substrates. The extent of fractionation depended on which enzymatic pathways were used, the direction in which the pathways operated, and the growth rate, but fractionation was not dependent on the growth phase. To the extent that environmental conditions affect the availability of organic substrates (e.g., acetate) and reducing power (e.g., H(2)), ecological forces can also influence carbon isotope discrimination by SRB. PMID- 12732571 TI - Sequence versus structure for the direct detection of 16S rRNA on planar oligonucleotide microarrays. AB - A two-probe proximal chaperone detection system consisting of a species-specific capture probe for the microarray and a labeled, proximal chaperone probe for detection was recently described for direct detection of intact rRNAs from environmental samples on oligonucleotide arrays. In this study, we investigated the physical spacing and nucleotide mismatch tolerance between capture and proximal chaperone detector probes that are required to achieve species-specific 16S rRNA detection for the dissimilatory metal and sulfate reducer 16S rRNAs. Microarray specificity was deduced by analyzing signal intensities across replicate microarrays with a statistical analysis-of-variance model that accommodates well-to-well and slide-to-slide variations in microarray signal intensity. Chaperone detector probes located in immediate proximity to the capture probe resulted in detectable, nonspecific binding of nontarget rRNA, presumably due to base-stacking effects. Species-specific rRNA detection was achieved by using a 22-nt capture probe and a 15-nt detector probe separated by 10 to 14 nt along the primary sequence. Chaperone detector probes with up to three mismatched nucleotides still resulted in species-specific capture of 16S rRNAs. There was no obvious relationship between position or number of mismatches and within- or between-genus hybridization specificity. From these results, we conclude that relieving secondary structure is of principal concern for the successful capture and detection of 16S rRNAs on planar surfaces but that the sequence of the capture probe is more important than relieving secondary structure for achieving specific hybridization. PMID- 12732572 TI - Antimicrobial effects of mustard flour and acetic acid against Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella enterica serovar Typhimurium. AB - This study was designed to investigate the individual and combined effects of mustard flour and acetic acid in the inactivation of food-borne pathogenic bacteria stored at 5 and 22 degrees C. Samples were prepared to achieve various concentrations by the addition of acetic acid (0, 0.5, or 1%) along with mustard flour (0, 10, or 20%) and 2% sodium chloride (fixed amount). Acid-adapted three strain mixtures of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella enterica serovar Typhimurium strains (10(6) to 10(7) CFU/ml) were inoculated separately into prepared mustard samples stored at 5 and 22 degrees C, and samples were assayed periodically. The order of bacterial resistance, assessed by the time required for the nominated populations to be reduced to undetectable levels against prepared mustards at 5 degrees C, was S. enterica serovar Typhimurium (1 day) < E. coli O157:H7 (3 days) < L. monocytogenes (9 days). The food-borne pathogens tested were reduced much more rapidly at 22 degrees C than at 5 degrees C. There was no synergistic effect with regard to the killing of the pathogens tested with the addition of 0.5% acetic acid to the mustard flour (10 or 20%). Mustard in combination with 0.5% acetic acid had less bactericidal activity against the pathogens tested than did mustard alone. The reduction of E. coli O157:H7 and L. monocytogenes among the combined treatments on the same storage day was generally differentiated as follows: control < mustard in combination with 0.5% acetic acid < mustard alone < mustard in combination with 1% acetic acid < acetic acid alone. Our study indicates that acidic products may limit microbial growth or survival and that the addition of small amounts of acetic acid (0.5%) to mustard can retard the reduction of E. coli O157:H7 and L. monocytogenes. These antagonistic effects may be changed if mustard is used alone or in combination with >1% acetic acid. PMID- 12732573 TI - Characterization of two tetrachloroethene-reducing, acetate-oxidizing anaerobic bacteria and their description as Desulfuromonas michiganensis sp. nov. AB - Two tetrachlorethene (PCE)-dechlorinating populations, designated strains BB1 and BRS1, were isolated from pristine river sediment and chloroethene-contaminated aquifer material, respectively. PCE-to-cis-1,2-dichloroethene-dechlorinating activity could be transferred in defined basal salts medium with acetate as the electron donor and PCE as the electron acceptor. Taxonomic analysis based on 16S rRNA gene sequencing placed both isolates within the Desulfuromonas cluster in the delta subdivision of the Proteobacteria. PCE was dechlorinated at rates of at least 139 nmol min(-1) mg of protein(-1) at pH values between 7.0 and 7.5 and temperatures between 25 and 30 degrees C. Dechlorination also occurred at 10 degrees C. The electron donors that supported dechlorination included acetate, lactate, pyruvate, succinate, malate, and fumarate but not hydrogen, formate, ethanol, propionate, or sulfide. Growth occurred with malate or fumarate alone, whereas oxidation of the other electron donors depended strictly on the presence of fumarate, malate, ferric iron, sulfur, PCE, or TCE as an electron acceptor. Nitrate, sulfate, sulfite, thiosulfate, and other chlorinated compounds were not used as electron acceptors. Sulfite had a strong inhibitory effect on growth and dechlorination. Alternate electron acceptors (e.g., fumarate or ferric iron) did not inhibit PCE dechlorination and were consumed concomitantly. The putative fumarate, PCE, and ferric iron reductases were induced by their respective substrates and were not constitutively present. Sulfide was required for growth. Both strains tolerated high concentrations of PCE, and dechlorination occurred in the presence of free-phase PCE (dense non-aqueous-phase liquids). Repeated growth with acetate and fumarate as substrates yielded a BB1 variant that had lost the ability to dechlorinate PCE. Due to the 16S rRNA gene sequence differences with the closest relatives and the unique phenotypic characteristics, we propose that the new isolates are members of a new species, Desulfuromonas michiganensis, within the Desulfuromonas cluster of the Geobacteraceae. PMID- 12732574 TI - Enterolysin A, a cell wall-degrading bacteriocin from Enterococcus faecalis LMG 2333. AB - A novel antimicrobial protein, designated enterolysin A, was purified from an Enterococcus faecalis LMG 2333 culture. Enterolysin A inhibits growth of selected enterococci, pediococci, lactococci, and lactobacilli. Antimicrobial activity was initially detected only on solid media, but by growing the bacteria in a fermentor under optimized production conditions (MRS broth with 4% [wt/vol] glucose, pH 6.5, and a temperature between 25 and 35 degrees C), the bacteriocin activity was increased to 5,120 bacteriocin units ml(-1). Enterolysin A production was regulated by pH, and activity was first detected in the transition between the logarithmic and stationary growth phases. Killing of sensitive bacteria by enterolysin A showed a dose-response behavior, and the bacteriocin has a bacteriolytic mode of action. Enterolysin A was purified, and the primary structure was determined by combined amino acid and DNA sequencing. This bacteriocin is translated as a 343-amino-acid preprotein with an sec-dependent signal peptide of 27 amino acids, which is followed by a sequence corresponding to the N-terminal part of the purified protein. Mature enterolysin A consists of 316 amino acids and has a calculated molecular weight of 34,501, and the theoretical pI is 9.24. The N terminus of enterolysin A is homologous to the catalytic domains of different cell wall-degrading proteins with modular structures. These include lysostaphin, ALE-1, zoocin A, and LytM, which are all endopeptidases belonging to the M37 protease family. The N-terminal part of enterolysin A is linked by a threonine-proline-rich region to a putative C terminal recognition domain, which shows significant sequence identity to two bacteriophage lysins. PMID- 12732575 TI - Thermophily in the Geobacteraceae: Geothermobacter ehrlichii gen. nov., sp. nov., a novel thermophilic member of the Geobacteraceae from the "Bag City" hydrothermal vent. AB - Little is known about the microbiology of the "Bag City" hydrothermal vent, which is part of a new eruption site on the Juan de Fuca Ridge and which is notable for its accumulation of polysaccharide on the sediment surface. A pure culture, designated strain SS015, was recovered from a vent fluid sample from the Bag City site through serial dilution in liquid medium with malate as the electron donor and Fe(III) oxide as the electron acceptor and then isolation of single colonies on solid Fe(III) oxide medium. The cells were gram-negative rods, about 0.5 micro m by 1.2 to 1.5 micro m, and motile and contained c-type cytochromes. Analysis of the 16S ribosomal DNA (rDNA) sequence of strain SS015 placed it in the family Geobacteraceae in the delta subclass of the Proteobacteria. Unlike previously described members of the Geobacteraceae, which are mesophiles, strain SS015 was a thermophile and grew at temperatures of between 35 and 65 degrees C, with an optimum temperature of 55 degrees C. Like many previously described members of the Geobacteraceae, strain SS015 grew with organic acids as the electron donors and Fe(III) or nitrate as the electron acceptor, with nitrate being reduced to ammonia. Strain SS015 was unique among the Geobacteraceae in its ability to use sugars, starch, or amino acids as electron donors for Fe(III) reduction. Under stress conditions, strain SS015 produced copious quantities of extracellular polysaccharide, providing a model for the microbial production of the polysaccharide accumulation at the Bag City site. The 16S rDNA sequence of strain SS015 was less than 94% similar to the sequences of previously described members of the Geobacteraceae; this fact, coupled with its unique physiological properties, suggests that strain SS015 represents a new genus in the family Geobacteraceae. The name Geothermobacter ehrlichii gen. nov., sp. nov., is proposed (ATCC BAA-635 and DSM 15274). Although strains of Geobacteraceae are known to be the predominant Fe(III)-reducing microorganisms in a variety of Fe(III)-reducing environments at moderate temperatures, strain SS015 represents the first described thermophilic member of the Geobacteraceae and thus extends the known environmental range of this family to hydrothermal environments. PMID- 12732576 TI - Community-level physiological profiling performed with an oxygen-sensitive fluorophore in a microtiter plate. AB - Community-level physiological profiling based upon fluorometric detection of oxygen consumption was performed on hydroponic rhizosphere and salt marsh litter samples by using substrate levels as low as 50 ppm with incubation times between 5 and 24 h. The rate and extent of response were increased in samples acclimated to specific substrates and were reduced by limiting nitrogen availability in the wells. PMID- 12732577 TI - Reductive dehalogenation of chlorobenzene congeners in cell extracts of Dehalococcoides sp. strain CBDB1. AB - Enzymatic reductive dehalogenation of tri-, tetra-, penta-, and hexachlorobenzenes was demonstrated in cell extracts with low protein concentration (0.5 to 1 micro g of protein/ml) derived from the chlorobenzene respiring anaerobe Dehalococcoides sp. strain CBDB1. 1,2,3-trichlorobenzene dehalogenase activity was associated with the membrane fraction. Light-reversible inhibition by alkyl iodides indicated the presence of a corrinoid cofactor. PMID- 12732578 TI - Endotoxin inactivation in water by using medium-pressure UV lamps. AB - Deionized water was spiked with various concentrations of endotoxin and exposed to UV irradiation from medium-pressure UV lamps to assess endotoxin inactivation. It was found that endotoxin inactivation was proportional to the UV dose under the conditions examined. The inactivation rate was determined to be approximately 0.55 endotoxin unit/ml per mJ/cm(2) of irradiation delivered. PMID- 12732579 TI - Antimicrobial-resistant Campylobacter species from retail raw meats. AB - The antimicrobial susceptibilities of 378 Campylobacter isolates were determined. Resistance to tetracycline was the most common (82%), followed by resistance to doxycycline (77%), erythromycin (54%), nalidixic acid (41%), and ciprofloxacin (35%). Campylobacter coli displayed significantly higher rates of resistance to ciprofloxacin and erythromycin than Campylobacter jejuni, and Campylobacter isolates from turkey meat showed a greater resistance than those from chicken meat. PMID- 12732581 TI - Production of native-type Streptoverticillium mobaraense transglutaminase in Corynebacterium glutamicum. AB - We previously observed secretion of active-form transglutaminase in Corynebacterium glutamicum by coexpressing the subtilisin-like protease SAM-P45 from Streptomyces albogriseolus to process the prodomain. However, the N-terminal amino acid sequence of the transglutaminase differed from that of the native Streptoverticillium mobaraense enzyme. In the present work we have used site directed mutagenesis to generate an optimal SAM-P45 cleavage site in the C terminal region of the prodomain. As a result, native-type transglutaminase was secreted. PMID- 12732582 TI - Unexpected thermal destruction of dried, glass bead-immobilized microorganisms as a function of water activity. AB - To help us understand the factors and mechanisms implicated in the death of microorganisms or their resistance to temperature in a low water activity environment, microorganisms were dried on the surface of glass beads before being subjected to high temperatures for a short period followed by rapid cooling. Two microorganisms were studied: the yeast Saccharomyces cerevisiae and the bacterium Lactobacillus plantarum. Experiments were carried out at 150, 200, and 250 degrees C, with four durations of heat treatment and seven levels of initial water activity between 0.10 and 0.70. We observed an unexpected range of water activity, between 0.30 and 0.50, at which microorganisms were more resistant to the various treatments, with maximal viability at 0.35 for L. plantarum and 0.40 for S. cerevisiae. PMID- 12732580 TI - Habitat-specific diversity of Borrelia burgdorferi sensu lato in Europe, exemplified by data from Latvia. AB - The distribution of Borrelia burgdorferi sensu lato genospecies in questing Ixodes ricinus ticks from ecologically distinct habitats in Latvia was analyzed. A significant variation in the frequency of the genospecies across sites was observed, pointing to the importance of the host community in the ecology of Lyme borreliosis. PMID- 12732583 TI - An improved cloning vector for construction of gene replacements in Listeria monocytogenes. AB - Listeria monocytogenes is a gram-positive, facultative intracellular bacterium implicated in severe food-borne illness (listeriosis) in humans. The construction of well-defined gene replacements in the genome of L. monocytogenes has been instrumental to several genetic studies of the virulence and other attributes of the organism. Construction of such mutations by currently available procedures, however, tends to be labor intensive, and gene replacement mutants are sometimes difficult to recover due to lack of direct selection for the construct. In this study we describe the construction and use of plasmid vector pGF-EM, which can be conjugatively transferred from Escherichia coli S17-1 to L. monocytogenes and which provides the genetic means for direct selection of gene replacements. PMID- 12732584 TI - Regression of isoproterenol-induced cardiac hypertrophy by Na+/H+ exchanger inhibition. AB - Cardiac hypertrophy is often associated with an increased sympathetic drive, and both in vitro and in vivo studies have demonstrated the development of cardiomyocyte hypertrophy in response to either alpha- or beta-adrenergic stimulation. Because an association between the Na+/H+ exchanger and cellular growth has been proposed, this study aimed to analyze the possible role of the antiporter in isoproterenol-induced cardiac hypertrophy. Isoproterenol alone (5 mg/kg IP once daily) or combined with a selective inhibitor of the Na+/H+ exchanger activity (3 mg x kg(-1) x d(-1) BIIB723) was given to male Wistar rats for 30 days. Sex- and age-matched rats that received 0.9% saline IP daily served as controls. Echocardiographic follow-up showed a 33% increase in left ventricular mass in the isoproterenol-treated group, whereas it did not increase in the isoproterenol+BIIB723-treated group. Heart weight-to-body weight ratio at necropsy was 2.44+/-0.11 in controls and increased to 3.35+/-0.10 (P<0.05) with isoproterenol, an effect that was markedly attenuated by BIIB723 (2.82+/-0.07). Intense cardiomyocyte enlargement and severe subendocardial fibrosis were found in isoproterenol-treated rats, and both effects were attenuated by BIIB723. Myocardial Na+/H+ exchanger activity and protein expression significantly increased in isoproterenol-treated rats compared with the control group (1.45+/ 0.11 vs 0.91+/-0.05 arbitrary units, P<0.05). This effect was significantly reduced by BIIB723 (1.17+/-0.02, P<0.05). In conclusion, our results show that Na+/H+ exchanger inhibition prevented the development of isoproterenol-induced hypertrophy and fibrosis, providing strong evidence in favor of a key role played by the antiporter in this model of cardiac hypertrophy. PMID- 12732585 TI - Baroreflex buffering in sedentary and endurance exercise-trained healthy men. AB - Baroreflex buffering plays an important role in arterial blood pressure control. Previous reports suggest that baroreflex sensitivity may be altered in endurance exercise-trained compared with untrained subjects. It is unknown, however, if in vivo baroreflex buffering is altered in the endurance exercise-trained state in humans. Baroreflex buffering was determined in 36 healthy normotensive men (18 endurance exercise-trained, 41+/-5 [SEM] years; 18 untrained, 41+/-4 years) by measuring the potentiation of the systolic blood pressure responses to a phenylephrine bolus and to incremental phenylephrine infusion during compared with before ganglionic blockade with trimethaphan. The exercise-trained men had a lower resting heart rate and higher maximal oxygen consumption and heart rate variability than the sedentary control subjects (all P=0.01). Mean levels and variability of blood pressure, cardiovagal baroreflex sensitivity (change in heart rate/change in systolic blood pressure), and basal muscle sympathetic nerve activity were not different in the two groups. The systolic blood pressure responses to phenylephrine were not different in the endurance-trained and untrained men before or during ganglionic blockade (P>0.6). Measures of baroreflex buffering with the use of a phenylephrine bolus (3.9+/-0.8 versus 4.0+/-0.7, trained versus untrained, P=0.85) and incremental infusion (2.8+/-0.4 versus 2.5+/-0.6, P=0.67) were similar in the two groups. Baroreflex buffering does not differ in endurance exercise-trained compared with untrained healthy men. These results support the concept that habitual vigorous endurance exercise does not modulate in vivo baroreflex buffering in healthy humans. PMID- 12732586 TI - Administration time-dependent effects of aspirin on blood pressure in untreated hypertensive patients. AB - Previous studies on the potential influence of aspirin on blood pressure have not taken into consideration the chronopharmacological effects of nonsteroidal anti inflammatory drugs. This pilot study investigates the effects of aspirin on blood pressure in untreated hypertensive patients who received aspirin at different times of the day according to their rest-activity cycle. We studied 100 untreated patients with mild hypertension (34 men and 66 women), 42.5+/-11.6 (mean+/-SD) years of age, randomly divided into 3 groups: nonpharmacological hygienic-dietary recommendations; the same recommendations and aspirin (100 mg/d) on awakening; or the same recommendations and aspirin before bedtime. Blood pressure was measured every 20 minutes during the day and every 30 minutes at night for 48 consecutive hours before and after 3 months of intervention. The circadian pattern of blood pressure in each group was established by population multiple-component analysis. After 3 months of nonpharmacological intervention, there was a small, nonsignificant reduction of blood pressure (<1.1 mm Hg; P>0.341). There was no change in blood pressure when aspirin was given on awakening (P=0.229). A highly significant blood pressure reduction was, however, observed in the patients who received aspirin before bedtime (decrease of 6 and 4 mm Hg in systolic and diastolic blood pressure, respectively; P<0.001). Results indicate a statistically significant administration time-dependent effect of low-dose aspirin on blood pressure in untreated patients with mild hypertension. The influence of aspirin on blood pressure demonstrated in this study indicates the need to quantify and control for aspirin effects in patients using this drug in combination with antihypertensive medication. PMID- 12732587 TI - Subcellular redistribution of focal adhesion kinase and its related nonkinase in hypertrophic myocardium. AB - Focal adhesion kinase (FAK) and focal adhesion kinase-related nonkinase (FRNK) are likely involved in mechanical signaling during hypertension. We investigated expression, subcellular distribution, and phosphorylation of FAK, as well as FRNK in left ventricles of spontaneously hypertensive heart failure rats. Compared with normotensive controls, FAK and FRNK increased in left ventricles of hypertensive rats. Increased FAK and FRNK were mainly present in membrane cytoskeleton and nuclear fractions. Confocal microscopy demonstrated that FAK and FRNK translocated to nuclei and intercalated disks in cardiac myocytes from hypertensive rats. Serine and tyrosine phosphorylation of FAK increased dramatically in hypertensive rats. FAK phosphorylated at tyrosine 397 was present in membranes and intercalated disks, but not in nuclei. FAK was also phosphorylated on serine 722 but not on serine 910. In contrast, FRNK was phosphorylated on serine 217, the equivalent site of FAK serine 910, but not serine on 30, the homologous site of FAK serine 722. Serine phosphorylated FAK and FRNK accumulated in membranes and nuclei but not in intercalated disks. Nuclear translocation of FAK and FRNK may play important roles in regulating mechanical signal transduction in cardiac myocytes. PMID- 12732588 TI - Ca2+ mobilization induced by ouabain in thymocytes involves intracellular and extracellular Ca2+ pools. AB - Immune dysfunction has been reported in hypertensive rats, and circulating levels of ouabain are increased in some experimental models of hypertension. Ouabain is an inhibitor of the Na+/K+-ATPase capable of diverse effects on cells of the immune system, but its mode of action on these cells is still unknown. The levels of cytoplasmic calcium ions play an important role in cell signaling, and ouabain may induce an increase in intracellular calcium indirectly through the Na+/Ca2+ exchanger. The current work examined the possibility that this drug could be exerting its effects on thymocytes through calcium mobilization and an increase in the cytosolic calcium concentration. Intracellular calcium was evaluated by using Balb-c mouse thymocytes loaded with FURA-2. Both intracellular and extracellular calcium pools were mobilized by ouabain (3 to 1000 nmol). The influx of extracellular calcium depended on the Na+/Ca2+ exchanger and on voltage dependent calcium channels, as it was inhibited by amiloride and benzamil, consistent with the inhibition of the Na+/K+ pump. In addition, the increase of calcium from intracellular stores was extremely rapid. Furthermore, an increase in cytosolic calcium levels was obtained with the combination of ouabain and thapsigargin, which was greater than that seen with either drug alone. Our data suggest that low concentrations of ouabain may be acting on thymocytes through a mechanism different from the traditional inhibition of the Na+/K+-ATPase, as the cytosolic calcium rise was partly dependent on the release from intracellular stores. PMID- 12732589 TI - Adolescent pain sensitivity is associated with cardiac autonomic function and blood pressure over 8 years. AB - Low pain sensitivity has been reported in hypertensive subjects as well in groups deemed to be at increased risk of development of the disorder. However, it is uncertain whether individual differences in pain sensitivity are associated prospectively with increases in blood pressure. In the current study, 24-hour blood pressure and heart rate variability were recorded in 110, 22-year-old men previously assessed at age 14 years for casual blood pressure and pain sensitivity (mechanical finger pressure). Significant correlations were observed between pain tolerance in 14-year-olds and current 24-hour systolic blood pressure (r=0.37, P<0.01) and diastolic blood pressure (r=0.36, P<0.01). Hierarchical multiple regression analyses indicated that information regarding pain tolerance improved prediction of systolic and diastolic blood pressure at age 22 years beyond that afforded by differences in blood pressure, parental history of hypertension, and body mass index at age 14 years. Similar analyses revealed that average pain sensitivity at age 14 was also associated with 24-hour high-frequency heart rate variability (r=0.28, P<0.01) and low-frequency/high frequency heart rate variability at age 22 (r=-0.35, P<0.01), suggesting increased sympathetic and reduced parasympathetic tone among individuals less sensitive to pain. These results provide further evidence that blood pressure related hypoalgesia might be related to processes involved in blood pressure regulation as well as in the development of sustained high blood pressure. PMID- 12732590 TI - Fifty years of open-heart surgery. PMID- 12732591 TI - To screen or not is not the question--it is when and how to screen. PMID- 12732592 TI - Inflammation, neointimal hyperplasia, and restenosis: as the leukocytes roll, the arteries thicken. PMID- 12732593 TI - Membrane microdomains and vascular biology: emerging role in atherogenesis. PMID- 12732594 TI - New frontiers in cardiology: drug-eluting stents: Part I. PMID- 12732595 TI - Images in cardiovascular medicine. Huge calcified pulmonary arterial aneurysm. PMID- 12732596 TI - Cardiology patient pages. Diagnostic cardiac catheterization. PMID- 12732597 TI - Images in cardiovascular medicine. Unusual biventricular thrombus formation in acute myeloid leukemia and factor V Leiden mutation. PMID- 12732599 TI - Evidence for a major quantitative trait locus on chromosome 17q21 affecting low density lipoprotein peak particle diameter. AB - BACKGROUND: Several lines of evidence suggest that small dense LDL particles are associated with the risk of coronary heart disease. Heritability and segregation studies suggest that LDL particle size is characterized by a large genetic contribution and the presence of a putative major genetic locus. However, association and linkage analyses have thus far been inconclusive in identifying the underlying gene(s). METHODS AND RESULTS: An autosomal genome-wide scan for LDL peak particle diameter (LDL-PPD) was performed in the Quebec Family Study. A total of 442 markers were genotyped, with an average intermarker distance of 7.2 cM. LDL-PPD was measured by gradient gel electrophoresis in 681 subjects from 236 nuclear families. Linkage was tested by both sib-pair-based and variance components-based linkage methods. The strongest evidence of linkage was found on chromosome 17q21.33 at marker D17S1301, with an LOD score of 6.76 by the variance components method for the phenotype adjusted for age, body mass index, and triglyceride levels. Similar results were obtained with the sib-pair method (P<0.0001). Other chromosomal regions harboring markers with highly suggestive evidence of linkage (P< or =0.0023; LOD > or =1.75) include 1p31, 2q33.2, 4p15.2, 5q12.3, and 14q31. Several candidate genes are localized under the peak linkages, including apolipoprotein H on chromosome 17q, the apolipoprotein E receptor 2, and members of the phospholipase A2 family on chromosome 1p as well as HMG-CoA reductase on chromosome 5q. CONCLUSIONS: This genome-wide scan for LDL-PPD indicates the presence of a major quantitative trait locus located on chromosome 17q and others interesting loci influencing the phenotype. PMID- 12732600 TI - Arginine vasopressin in advanced vasodilatory shock: a prospective, randomized, controlled study. AB - BACKGROUND: Vasodilatory shock is a potentially lethal complication of severe disease in critically ill patients. Currently, catecholamines are the most widely used vasopressor agents to support blood pressure, but loss of catecholamine pressor effects is a well-known clinical dilemma. Arginine vasopressin (AVP) has recently been shown to be a potent vasopressor agent to stabilize cardiocirculatory function even in patients with catecholamine-resistant vasodilatory shock. METHODS AND RESULTS: Forty-eight patients with catecholamine resistant vasodilatory shock were prospectively randomized to receive a combined infusion of AVP and norepinephrine (NE) or NE infusion alone. In AVP patients, AVP was infused at a constant rate of 4 U/h. Hemodynamic, acid/base, single organ, and tonometrically derived gastric variables were reported before the study and 1, 12, 24, and 48 hours after study entry. For statistical analysis, a mixed-effects model was used. AVP patients had significantly lower heart rate, NE requirements, and incidence of new-onset tachyarrhythmias than NE patients. Mean arterial pressure, cardiac index, stroke volume index, and left ventricular stroke work index were significantly higher in AVP patients. NE patients developed significantly more new-onset tachyarrhythmias than AVP patients (54.3% versus 8.3%). Gastrointestinal perfusion as assessed by gastric tonometry was better preserved in AVP-treated patients. Total bilirubin concentrations were significantly higher in AVP patients. CONCLUSIONS: The combined infusion of AVP and NE proved to be superior to infusion of NE alone in the treatment of cardiocirculatory failure in catecholamine-resistant vasodilatory shock. PMID- 12732601 TI - Treatment of saphenous vein bypass grafts with ultrasound thrombolysis: a randomized study (ATLAS). AB - BACKGROUND: Percutaneous coronary interventions (PCIs) in saphenous vein grafts (SVGs) with thrombus have a high frequency of distal embolization. Acolysis (therapeutic ultrasound) can break up thrombus in vitro in animal models and humans. Whether this is beneficial during percutaneous SVG interventions is unknown. METHODS AND RESULTS: We performed a trial of coronary ultrasound thrombolysis in which patients with an acute coronary syndrome undergoing PCI in SVGs were randomly assigned to receive acolysis or abciximab. The primary end point was a successful procedure, defined as final luminal diameter stenosis 30% or less with Thrombolysis In Myocardial Infarction grade 3 flow and freedom from major adverse cardiac events (composite of death, Q-wave, and non-Q-wave myocardial infarction [MI], emergency bypass procedure, disabling stroke, and target lesion revascularization). Of 181 enrolled, 92 received acolysis and 89 abciximab. Angiographic procedural success was achieved in 63% of acolysis patients and 82% of abciximab patients (P=0.008). Incidence of major adverse cardiac events at 30 days was 25% with acolysis and 12% with abciximab (P=0.036), attributable mainly to a greater frequency of non-Q-wave MI with acolysis (19.6% versus 7.9%, P=0.03). The incidence of Q-wave MI was also higher with acolysis (5.4% versus 2.2%, P=nonsignificant). The primary end point was achieved in 53.8% of acolysis patients and 73.1% of abciximab patients (P=0.014). CONCLUSIONS: Use of therapeutic ultrasound in vein graft lesions in patients with acute coronary syndrome had poor angiographic outcome and increased the incidence of acute ischemic complications. PMID- 12732602 TI - Chronic iron administration increases vascular oxidative stress and accelerates arterial thrombosis. AB - BACKGROUND: Iron overload has been implicated in the pathogenesis of ischemic cardiovascular events. However, the effects of iron excess on vascular function and the thrombotic response to vascular injury are not well understood. METHODS AND RESULTS: We examined the effects of chronic iron dextran administration (15 mg over 6 weeks) on thrombosis, systemic and vascular oxidative stress, and endothelium-dependent vascular reactivity in mice. Thrombus generation after photochemical carotid artery injury was accelerated in iron-loaded mice (mean time to occlusive thrombosis, 20.4+/-8.5 minutes; n=10) compared with control mice (54.5+/-35.5 minutes, n=10, P=0.009). Iron loading had no effect on plasma clotting, vessel wall tissue factor activity, or ADP-induced platelet aggregation. Acute administration of dl-cysteine, a reactive oxygen species scavenger, completely abrogated the effects of iron loading on thrombus formation, suggesting that iron accelerated thrombosis through a pro-oxidant mechanism. Iron loading enhanced both systemic and vascular reactive oxygen species production. Endothelium-dependent vasorelaxation was impaired in iron loaded mice, indicating reduced NO bioavailability. CONCLUSIONS: Moderate iron loading markedly accelerates thrombus formation after arterial injury, increases vascular oxidative stress, and impairs vasoreactivity. Iron-induced vascular dysfunction may contribute to the increased incidence of ischemic cardiovascular events that have been associated with chronic iron overload. PMID- 12732603 TI - Prevention of chronic deterioration of heart allograft by recombinant adeno associated virus-mediated heme oxygenase-1 gene transfer. AB - BACKGROUND: Allograft deterioration is the major obstacle to organ transplantation as a long-term treatment of end-stage heart failure. In this study, we transduced the antioxidant gene, heme oxygenase-1 (HO-1), to heart grafts using a recombinant adeno-associated viral vector (rAAV) in a rat heart transplantation model and investigated its potentiality in prevention of chronic graft deterioration. METHODS AND RESULTS: rAAV/HO-1 was administered to heart grafts through the coronary arteries during cold preservation. We investigated the expression patterns and activities of transgene, graft survival, graft histomorphology, and relevance of HO-1 expression on graft survival and chronic graft deterioration by itself. Long-term allograft survival can be achieved by rAAV/HO-1-mediated stable transgene expression. The development of graft arteriosclerosis and interstitial fibrosis was prevented in rAAV/HO-1-transduced allografts on day 100. rAAV/HO-1-mediated long-term graft protection was accompanied by remarkable downregulation of the intragraft mRNA level of macrophage migration inhibitory factor, tumor necrosis factor-alpha, and transforming growth factor-beta1. Blockage of HO activities by zinc protoporphyrin IX at different posttransplant phases showed that the stable expression of HO-1 is a prerequisite for both survival of grafts and prevention of graft arteriosclerosis. CONCLUSIONS: rAAV/HO-1 gene transfer represents a novel therapeutic approach to prevent chronic allograft deterioration in clinical heart transplantation. PMID- 12732604 TI - Alterations in atrial electrophysiology and tissue structure in a canine model of chronic atrial dilatation due to mitral regurgitation. AB - BACKGROUND: Clinically, chronic atrial dilatation is associated with an increased incidence of atrial fibrillation (AF), but the underlying mechanism is not clear. We have investigated atrial electrophysiology and tissue structure in a canine model of chronic atrial dilatation due to mitral regurgitation (MR). METHODS AND RESULTS: Thirteen control and 19 MR dogs (1 month after partial mitral valve avulsion) were studied. Dogs in the MR group were monitored using echocardiography and Holter recording. In open-chest follow-up experiments, electrode arrays were placed on the atria to investigate conduction patterns, effective refractory periods, and inducibility of AF. Alterations in tissue structure and ultrastructure were assessed in atrial tissue samples. At follow up, left atrial length in MR dogs was 4.09+/-0.45 cm, compared with 3.25+/-0.28 at baseline (P<0.01), corresponding to a volume of 205+/-61% of baseline. At follow-up, no differences in atrial conduction pattern and conduction velocities were noted between control and MR dogs. Effective refractory periods were increased homogeneously throughout the left and right atrium. Sustained AF (>1 hour) was inducible in 10 of 19 MR dogs and none of 13 control dogs (P<0.01). In the dilated MR left atrium, areas of increased interstitial fibrosis and chronic inflammation were accompanied by increased glycogen ultrastructurally. CONCLUSIONS: Chronic atrial dilatation in the absence of overt heart failure leads to an increased vulnerability to AF that is not based on a decrease in wavelength. PMID- 12732605 TI - Immediate administration of mineralocorticoid receptor antagonist spironolactone prevents post-infarct left ventricular remodeling associated with suppression of a marker of myocardial collagen synthesis in patients with first anterior acute myocardial infarction. AB - BACKGROUND: Aldosterone (ALD) has been shown to stimulate cardiac collagen synthesis and fibroblast proliferation via activation of local mineralocorticoid receptors. In patients with acute myocardial infarction, we demonstrated that ALD was extracted through the infarct heart and extracting ALD-stimulated post infarct left ventricular (LV) remodeling. METHODS AND RESULTS: To evaluate the effect of mineralocorticoid receptor antagonist (MRA) spironolactone on post infarct LV remodeling, 134 patients with first anterior acute myocardial infarction were randomly divided into the MRA (n=65) or non-MRA (n=69) groups after revascularization. All patients were administered angiotensin-converting enzyme (ACE) inhibitor and study drug just after revascularization. Left ventriculography with contrast medium was performed at the acute stage and after 1 month to evaluate LV remodeling. ALD was measured at aortic root and coronary sinus. There was no difference in the baseline characteristics including infarct size and LV performance between the two groups. However, LV ejection fraction was significantly improved in the MRA group compared with that in the non-MRA group (46.0+/-0.6% to 53.2+/-0.8% versus 46.5+/-0.8% to 51.0+/-0.8%, Pinteraction=0.012). LV end-diastolic volume index was significantly suppressed in the MRA group compared with that in non-MRA group (86.5+/-1.0 to 90.6+/-2.4 versus 87.5+/-1.3 to 106.8+/-3.5 mL/m2, Pinteraction=0.002). Transcardiac extraction of ALD through the heart was significantly suppressed in the MRA group (Pinteraction=0.001), and plasma procollagen type III aminoterminal peptide level, a biochemical marker of fibrosis, was significant lower in the MRA group compared with the non-MRA group (Pinteraction=0.002). CONCLUSIONS: These findings indicate that MRA combined with ACE inhibitor can prevent post-infarct LV remodeling better than ACE inhibitor alone in association with the suppression of a marker of collagen synthesis. PMID- 12732607 TI - Electrical resynchronization: a novel therapy for the failing right ventricle. AB - BACKGROUND: Many patients with congenital heart disease develop right ventricular (RV) failure due to anatomy and prior therapy. RV problems may include right bundle-branch block (RBBB), volume loading, and chamber enlargement. Because the failing RV may have regional dyskinesis, we hypothesized that resynchronization therapy might augment its performance. METHODS AND RESULTS: We studied 7 patients with RV dysfunction and RBBB, using a predefined pacing protocol. QRS duration, cardiac index (CI), and RV dP/dt were measured in 4 different pacing states. Atrioventricular pacing improved CI and RV dP/dtmax and decreased QRS duration as compared with atrial pacing or sinus rhythm. CONCLUSIONS: Atrioventricular pacing in patients with RBBB and RV dysfunction augments RV and systemic performance. RV resynchronization is a promising novel therapy for patients with RV failure. PMID- 12732606 TI - Left ventricular remodeling and ventricular arrhythmias after myocardial infarction. AB - BACKGROUND: The relation between left ventricular (LV) remodeling and ventricular arrhythmias after myocardial infarction is poorly documented. We investigated the relations between LV size, hypertrophy, and function and ventricular arrhythmias in 263 patients from the Survival and Ventricular Enlargement (SAVE) study, using quantitative 2D echocardiography and ambulatory ECG monitoring after myocardial infarction. METHODS AND RESULTS: Transthoracic 2D echocardiograms and arrhythmia monitoring were performed at baseline (mean, 11 days) and 1 and 2 years after infarction. LV size, short-axis muscle (mass) area (LVMA), and function were quantified from 2D echocardiograms. The prevalence of ventricular tachycardia (VT) and frequent ventricular ectopy (premature ventricular contractions [PVCs] >10/h) was assessed from ambulatory ECG. VT and PVCs >10/h occurred in 20% and 29% of patients at baseline, in 22% and 35% at 1 year and 23% and 39% at 2 years, respectively. VT and PVCs >10/h at baseline and 1 and 2 years were significantly related to LV size, LVMA, and function. Furthermore, changes in LV size and function from baseline to 2 years predicted both VT and PVCs >10/h. The study was underpowered to detect treatment effect of ACE inhibitors and beta-adrenergic receptor blockers but did not alter the relations between ventricular arrhythmias, LV size, and function. CONCLUSIONS: Quantitative echocardiographic assessment of LV size, LVMA, and function and changes in these measurements over time predict ventricular arrhythmias after infarction. Altered LV architecture and function during postinfarction LV remodeling provide an important substrate for triggering high-grade ventricular arrhythmias. PMID- 12732608 TI - In vivo magnetic resonance imaging of mesenchymal stem cells in myocardial infarction. AB - BACKGROUND: We investigated the potential of magnetic resonance imaging (MRI) to track magnetically labeled mesenchymal stem cells (MR-MSCs) in a swine myocardial infarction (MI) model. METHODS AND RESULTS: Adult farm pigs (n=5) were subjected to closed-chest experimental MI. MR-MSCs (2.8 to 16x107 cells) were injected intramyocardially under x-ray fluoroscopy. MRIs were obtained on a 1.5T MR scanner to demonstrate the location of the MR-MSCs and were correlated with histology. Contrast-enhanced MRI demonstrated successful injection in the infarct and serial MSC tracking was demonstrated in two animals. CONCLUSIONS: MRI tracking of MSCs is feasible and represents a preferred method for studying the engraftment of MSCs in MI. PMID- 12732609 TI - Angiotensin II type 2 receptor deficiency exacerbates heart failure and reduces survival after acute myocardial infarction in mice. AB - BACKGROUND: Angiotensin II plays a prominent role in the progression of heart failure after acute myocardial infarction (AMI). Although both angiotensin type 1 (AT1) and type 2 (AT2) receptors are known to be present in the heart, comparatively little is known about the latter. We therefore examined the role played by AT2 receptors in post-AMI heart failure. METHODS AND RESULTS: In wild type mice subjected to AMI by coronary artery ligation, AT2 receptor immunoreactivity is upregulated in the infarct and border areas. Among AT2 receptor-null (-/-) mice, the 7-day survival rate after AMI was significantly lower than among wild-type mice (43% versus 67%; P<0.05). All sham-operated animals of both genotypes survived through the study. Ventricular mRNA levels for brain natriuretic peptide were elevated in both genotypes 24 hours after coronary occlusion, with levels in AT2-/- significantly higher than in wild-type mice, as were their lung weights, and histological examination revealed marked pulmonary congestion in the AT2-/- mice. Cardiac function was significantly decreased in AT2-/- mice 2 days after AMI. CONCLUSIONS: AT2 receptor deficiency exacerbates short-term death rates and heart failure after experimental AMI in mice. The AT2 receptor may thus exert a protective effect on the heart after AMI. PMID- 12732610 TI - L-4F, an apolipoprotein A-1 mimetic, dramatically improves vasodilation in hypercholesterolemia and sickle cell disease. AB - BACKGROUND: Hypercholesterolemia and sickle cell disease (SCD) impair endothelium dependent vasodilation by dissimilar mechanisms. Hypercholesterolemia impairs vasodilation by a low-density lipoprotein (LDL)-dependent mechanism. SCD has been characterized as a chronic state of inflammation in which xanthine oxidase (XO) from ischemic tissues increases vascular superoxide anion (O2*-) generation. Recent reports indicate that apolipoprotein (apo) A-1 mimetics inhibit atherosclerosis in LDL receptor-null (Ldlr-/-) mice fed Western diets. Here we hypothesize that L-4F, an apoA-1 mimetic, preserves vasodilation in hypercholesterolemia and SCD by decreasing mechanisms that increase O2*- generation. METHODS AND RESULTS: Arterioles were isolated from hypercholesterolemic Ldlr-/- mice and from SCD mice that were treated with either saline or L-4F (1 mg/kg per day). Vasodilation in response to acetylcholine was determined by videomicroscopy. Effects of L-4F on LDL-induced increases in endothelium-dependent O2*- generation were determined on arterial segments via the hydroethidine assay and on stimulated endothelial cell cultures via superoxide dismutase-inhibitable ferricytochrome c reduction. Effects of L-4F on XO bound to pulmonary arterioles and content in livers of SCD mice were determined by immunofluorescence. Hypercholesterolemia impaired vasodilation in Ldlr-/- mice, which L-4F dramatically improved. L-4F inhibited LDL-induced increases in O2*- in arterial segments and in stimulated cultures. SCD impaired vasodilation, increased XO bound to pulmonary endothelium, and decreased liver XO content. L-4F dramatically improved vasodilation, decreased XO bound to pulmonary endothelium, and increased liver XO content compared with levels in untreated SCD mice. CONCLUSIONS: These data show that L-4F protects endothelium-dependent vasodilation in hypercholesterolemia and SCD. Our findings suggest that L-4F restores vascular endothelial function in diverse models of disease and may be applicable to treating a variety of vascular diseases. PMID- 12732611 TI - Leaflet escape in a new bileaflet mechanical valve: TRI technologies. AB - BACKGROUND: Leaflet escape is a mode of structural valve failure for mechanical prostheses. This complication previously has been reported for both monoleaflet and bileaflet valve models. We report 2 leaflet escape occurrences observed in 2 patients who underwent valve replacement with a TRI Technologies valve prosthesis. METHODS AND RESULTS: At the University of Padua, between November 2000 and February 2002, 36 TRI Technologies valve prostheses (26 aortic and 10 mitral) were implanted in 34 patients (12 women and 22 men) with a mean age of 59.9+/-10.3 years (range, 30 to 75 years). There were 5 deaths: 3 in hospital, 1 early after discharge, and 1 late. Two patients experienced a catastrophic prosthetic leaflet escape; the first patient was a 52-year-old man who died 10 days after aortic valve and ascending aorta replacement, and the second was a 58 year-old man who underwent a successful emergency reoperation 20 months after mitral valve replacement. Examination of the explanted prostheses showed in both cases a leaflet escape caused by a leaflet's pivoting system fracture. Prophylactic replacement was then successfully accomplished so far in 12 patients, without evidence of structural valve failure in any of them. Among other significant postoperative complications, we observed 3 major thromboembolisms, 1 hemorrhage, and 1 paravalvular leak. CONCLUSIONS: These catastrophes prompted us to interrupt the implantation program, and they cast a shadow on the durability of the TRI Technologies valve prosthesis because of its high risk of structural failure. PMID- 12732612 TI - American Society of Clinical Oncology technology assessment working group update: use of aromatase inhibitors in the adjuvant setting. PMID- 12732613 TI - Oscillatory nucleocytoplasmic shuttling of the general stress response transcriptional activators Msn2 and Msn4 in Saccharomyces cerevisiae. AB - Msn2 and Msn4 are two related transcriptional activators that mediate a general response to stress in yeast Saccharomyces cerevisiae by eliciting the expression of specific sets of genes. In response to stress or nutritional limitation, Msn2 and Msn4 migrate from the cytoplasm to the nucleus. Using GFP-tagged constructs and high-resolution time-lapse video microscopy on single cells, we show that light emitted by the microscope also triggers this migration. Unexpectedly, the population of Msn2 or Msn4 molecules shuttles repetitively into and out of the nucleus with a periodicity of a few minutes. A large heterogeneity in the oscillatory response to stress is observed between individual cells. This periodic behavior, which can be induced by various types of stress, at intermediate stress levels, is not dependent upon protein synthesis and persists when the DNA-binding domain of Msn2 is removed. The cAMP-PKA pathway controls the sensitivity of the oscillatory nucleocytoplasmic shuttling. In the absence of PKA, Msn4 continues to oscillate while Msn2 is maintained in the nucleus. We show that a computational model based on the possibility that Msn2 and Msn4 participate in autoregulatory loops controlling their subcellular localization can account for the oscillatory behavior of the two transcription factors. PMID- 12732614 TI - Proprotein convertase cleavage liberates a fibrillogenic fragment of a resident glycoprotein to initiate melanosome biogenesis. AB - Lysosome-related organelles are cell type-specific intracellular compartments with distinct morphologies and functions. The molecular mechanisms governing the formation of their unique structural features are not known. Melanosomes and their precursors are lysosome-related organelles that are characterized morphologically by intralumenal fibrous striations upon which melanins are polymerized. The integral membrane protein Pmel17 is a component of the fibrils and can nucleate their formation in the absence of other pigment cell-specific proteins. Here, we show that formation of intralumenal fibrils requires cleavage of Pmel17 by a furin-like proprotein convertase (PC). As in the generation of amyloid, proper cleavage of Pmel17 liberates a lumenal domain fragment that becomes incorporated into the fibrils; longer Pmel17 fragments generated in the absence of PC activity are unable to form organized fibrils. Our results demonstrate that PC-dependent cleavage regulates melanosome biogenesis by controlling the fibrillogenic activity of a resident protein. Like the pathologic process of amyloidogenesis, the formation of other tissue-specific organelle structures may be similarly dependent on proteolytic activation of physiological fibrillogenic substrates. PMID- 12732615 TI - A novel human protein of the maternal centriole is required for the final stages of cytokinesis and entry into S phase. AB - Centrosomes nucleate microtubules and contribute to mitotic spindle organization and function. They also participate in cytokinesis and cell cycle progression in ways that are poorly understood. Here we describe a novel human protein called centriolin that localizes to the maternal centriole and functions in both cytokinesis and cell cycle progression. Centriolin silencing induces cytokinesis failure by a novel mechanism whereby cells remain interconnected by long intercellular bridges. Most cells continue to cycle, reenter mitosis, and form multicellular syncytia. Some ultimately divide or undergo apoptosis specifically during the protracted period of cytokinesis. At later times, viable cells arrest in G1/G0. The cytokinesis activity is localized to a centriolin domain that shares homology with Nud1p and Cdc11p, budding and fission yeast proteins that anchor regulatory pathways involved in progression through the late stages of mitosis. The Nud1p-like domain of centriolin binds Bub2p, another component of the budding yeast pathway. We conclude that centriolin is required for a late stage of vertebrate cytokinesis, perhaps the final cell cleavage event, and plays a role in progression into S phase. PMID- 12732617 TI - Mitochondria to nucleus stress signaling: a distinctive mechanism of NFkappaB/Rel activation through calcineurin-mediated inactivation of IkappaBbeta. AB - Mitochondrial genetic and metabolic stress causes activation of calcineurin (Cn), NFAT, ATF2, and NFkappaB/Rel factors, which collectively alter the expression of an array of nuclear genes. We demonstrate here that mitochondrial stress-induced activation of NFkappaB/Rel factors involves inactivation of IkappaBbeta through Cn-mediated dephosphorylation. Phosphorylated IkappaBbeta is a substrate for Cn phosphatase, which was inhibited by FK506 and RII peptide. Chemical cross-linking and coimmunoprecipitation show that NFkappaB/Rel factor-bound IkappaBbeta forms a ternary complex with Cn under in vitro and in vivo conditions that was sensitive to FK506. Results show that phosphorylation at S313 and S315 from the COOH terminal PEST domain of IkappaBbeta is critical for binding to Cn. Mutations at S313/S315 of IkappaBbeta abolished Cn binding, inhibited Cn-mediated increase of Rel proteins in the nucleus, and had a dominant-negative effect on the mitochondrial stress-induced expression of RyR1 and cathepsin L genes. Our results show the distinctive nature of mitochondrial stress-induced NFkappaB/Rel activation, which is independent of IKKalpha and IKKbeta kinases and affects gene target(s) that are different from cytokine and TNFalpha-induced stress signaling. The results provide new insights into the role of Cn as a critical link between Ca2+ signaling and NFkappaB/Rel activation. PMID- 12732616 TI - Essential functions of p21-activated kinase 1 in morphogenesis and differentiation of mammary glands. AB - Although growth factors have been shown to influence mammary gland development, the nature of downstream effectors remains elusive. In this study, we show that the expression of p21-activated kinase (Pak)1, a serine/threonine protein kinase, is activated in mammary glands during pregnancy and lactation. By targeting an ectopic expression of a kinase-dead Pak1 mutant under the control of ovine beta lactoglobulin promoter, we found that the mammary glands of female mice expressing kinase-dead Pak1 transgene revealed incomplete lobuloalveolar development and impaired functional differentiation. The expression of whey acidic protein and beta-casein and the amount of activated Stat5 in the nuclei of epithelial cells in transgenic mice were drastically reduced. Further analysis of the underlying mechanisms revealed that Pak1 stimulated beta-casein promoter activity in normal mouse mammary epithelial cells and also cooperated with Stat5a. Pak1 directly interacted with and phosphorylated Stat5a at Ser 779, and both COOH-terminal deletion containing Ser 779 of Stat5a and the Ser 779 to Ala mutation completely prevented the ability of Pak1 to stimulate beta-casein promoter. Mammary glands expressing inactive Pak1 exhibited a reduction of Stat5a Ser 779 phosphorylation. These findings suggest that Pak1 is required for alveolar morphogenesis and lactation function, and thus, identify novel functions of Pak1 in the mammary gland development. PMID- 12732619 TI - Convergence of Wnt signaling and steroidogenic factor-1 (SF-1) on transcription of the rat inhibin alpha gene. AB - The action of a variety of peptide hormones is critical for proper growth and differentiation of the urogenital ridge, which ultimately gives rise to the kidney, adrenal cortex, and gonad. One such class of peptides is the Wnt family of secreted glycoproteins that is classically involved in development of cell polarity and cell fate determination. Notably, alterations in Wnt-4 expression in mice and humans result in profound defects in urogenital ridge development, including dysregulation of kidney, gonadal, and adrenal growth. The nuclear receptor steroidogenic factor-1 (SF-1) has been implicated as a downstream effector of peptide hormone signaling during urogenital ridge development as evidenced by both the activation of SF-1-dependent transcription in the adrenal cortex by signaling molecules such as protein kinase A and by the adrenal and gonadal agenesis in mice with null mutations in SF-1. We hypothesized that Wnt dependent signaling cascades regulate SF-1-dependent transcription of genes required for adreno-gonadal development. Specifically, the data demonstrate that beta-catenin synergizes with SF-1 to activate the alpha-inhibin promoter through formation of a transcriptional complex. The activation requires an intact SF-1 RE and is independent of TCF/Lef. These data support the recent observation that beta-catenin can participate in nuclear receptor-mediated transcriptional activation and extend the findings to the monomer binding class of orphan nuclear receptors. PMID- 12732618 TI - Purification, characterization, and molecular cloning of a novel keratan sulfate hydrolase, endo-beta-N-acetylglucosaminidase, from Bacillus circulans. AB - Keratan sulfate (KS) is degraded by various enzymes including endo-beta galactosidase, keratanase, and keratanase II, which are used for the structural analysis of KS. We purified a novel KS hydrolase, endo-beta-N acetylglucosaminidase, from the cell pellet and conditioned medium of Bacillus circulans, by sequential chromatography using DE52 and phenyl-Sepharose columns with approximately 63- and 180-fold purity and 58 and 12.5% recovery, respectively. Like keratanase II of Bacillus sp. Ks36, the enzyme, designated Bc keratanase II, hydrolyzed KS between the 4GlcNAcbeta1-3Gal1 structure (endo-beta N-acetylglucosaminidase), but not hyaluronan, heparan sulfate, heparin, and chondroitin sulfate C, demonstrating a strict specificity to KS. The enzyme digested shark cartilage KS to disaccharides and tetrasaccharides and bovine cornea KS to hexasaccharide, indicating that it prefers highly sulfated KS. Distinct from keratanase II of strain Ks36, the enzyme digested shark cartilage KS at an optimal temperature of 55 degrees C. Based on partial peptide sequencing of the enzyme, we molecularly cloned the gene, which encodes a protein with a predicted molecular mass of approximately 200 kDa. From the deduced protein sequence, Bc keratanase II contained a domain at the C terminus, homologous to the S-layer-like domain of pullulanase from Thermoanaerobacterium thermosulfurigenes and endoxylanase from Thermoanaerobacterium saccharolyticum, and a carbohydrate-binding domain, which may serve to specifically recognize KS chains. A full-length recombinant enzyme showed keratanase II activity. These results may prove useful for the structural analysis of KS toward achieving an understanding of its function. PMID- 12732620 TI - Missense, nonsense, and neutral mutations define juxtaposed regulatory elements of splicing in cystic fibrosis transmembrane regulator exon 9. AB - Exonic sequence variations may induce exon inclusion or exclusion from the mature mRNA by disrupting exonic regulatory elements and/or by affecting a nuclear reading frame scanning mechanism. We have carried out a systematic study of the effect on cystic fibrosis transmembrane regulator exon 9 splicing of natural and site-directed sequence mutations. We have observed that changes in the splicing pattern were not related to the creation of premature termination codons, a fact that indicates the lack of a significant nuclear check of the reading frame in this system. In addition, the splice pattern could not be predicted by available Ser/Arg protein matrices score analysis. An extensive site-directed mutagenesis of the 3' portion of the exon has identified two juxtaposed splicing enhancer and silencer elements. The study of double mutants at these regulatory elements showed a complex regulatory activity. For example, one natural mutation (146C) enhances exon inclusion and overrides all of the downstream silencing mutations except for a C to G transversion (155G). This unusual effect is explained by the creation of a specific binding site for the inhibitory splicing factor hnRNPH. In fact, on the double mutant 146C-155G, the silencing effect is dominant. These results indicate a strict dependence between the two juxtaposed enhancer and silencer sequences and show that many point mutations in these elements cause changes in splicing efficiency by different mechanisms. PMID- 12732621 TI - The putative catalytic bases have, at most, an accessory role in the mechanism of arginine kinase. AB - Arginine kinase is a member of the phosphagen kinase family that includes creatine kinase and likely shares a common reaction mechanism in catalyzing the buffering of cellular ATP energy levels. Abstraction of a proton from the substrate guanidinium by a catalytic base has long been thought to be an early mechanistic step. The structure of arginine kinase as a transition state analog complex (Zhou, G., Somasundaram, T., Blanc, E., Parthasarathy, G., Ellington, W. R., and Chapman, M. S. (1998) Proc. Natl. Acad. Sci. U. S. A. 95, 8449-8454) showed that Glu-225 and Glu-314 were the only potential catalytic residues contacting the phosphorylated nitrogen. In the present study, these residues were changed to Asp, Gln, and Val or Ala in several single and multisite mutant enzymes. These mutations had little impact on the substrate binding constants. The effect upon activity varied with reductions in kcat between 3000-fold and less than 2-fold. The retention of significant activity in some mutants contrasts with published studies of homologues and suggests that acid-base catalysis by these residues may enhance the rate but is not absolutely essential. Crystal structures of mutant enzymes E314D at 1.9 A and E225Q at 2.8 A resolution showed that the precise alignment of substrates is subtly distorted. Thus, pre-ordering of substrates might be just as important as acid-base chemistry, electrostatics, or other potential effects in the modest impact of these residues upon catalysis. PMID- 12732622 TI - Prion, amyloid beta-derived Cu(II) ions, or free Zn(II) ions support S-nitroso dependent autocleavage of glypican-1 heparan sulfate. AB - Copper are generally bound to proteins, e.g. the prion and the amyloid beta proteins. We have previously shown that copper ions are required to nitrosylate thiol groups in the core protein of glypican-1, a heparan sulfate-substituted proteoglycan. When S-nitrosylated glypican-1 is then exposed to an appropriate reducing agent, such as ascorbate, nitric oxide is released and autocatalyzes deaminative cleavage of the glypican-1 heparan sulfate side chains at sites where the glucosamines are N-unsubstituted. These processes take place in a stepwise manner, whereas glypican-1 recycles via a caveolin-1-associated pathway where copper ions could be provided by the prion protein. Here we show, by using both biochemical and microscopic techniques, that (a) the glypican-1 core protein binds copper(II) ions, reduces them to copper(I) when the thiols are nitrosylated and reoxidizes copper(I) to copper(II) when ascorbate releases nitric oxide; (b) maximally S-nitrosylated glypican-1 can cleave its own heparan sulfate chains at all available sites in a nitroxyl ion-dependent reaction; (c) free zinc(II) ions, which are redox inert, also support autocleavage of glypican-1 heparan sulfate, probably via transnitrosation, whereas they inhibit copper(II)-supported degradation; and (d) copper(II)-loaded but not zinc(II)-loaded prion protein or amyloid beta peptide support heparan sulfate degradation. As glypican-1 in prion null cells is poorly S-nitrosylated and as ectopic expression of cellular prion protein restores S-nitrosylation of glypican-1 in these cells, we propose that one function of the cellular prion protein is to deliver copper(II) for the S nitrosylation of recycling glypican-1. PMID- 12732623 TI - Formation of disulfide bonds and homodimers of the major cat allergen Fel d 1 equivalent to the natural allergen by expression in Escherichia coli. AB - Dander from the domestic cat (Felis domesticus) is one of the most common causes of IgE-mediated allergy. Attempts to produce tetrameric folded major allergen Fel d 1 by recombinant methods with structural features similar to the natural allergen have been only partially successful. In this study, a recombinant folded Fel d 1 with molecular and biological properties similar to the natural counterpart was produced. A synthetic gene coding for direct fusion of the Fel d 1 chain 2 N-terminally to chain 1 was constructed by overlapping oligonucleotides in PCR. Escherichia coli expression resulted in a non-covalently associated homodimer with an apparent molecular mass of 30 kDa defined by size exclusion chromatography. Furthermore, each 19,177-Da subunit displayed a disulfide pattern identical to that found in the natural Fel d 1, i.e. Cys3(1) Cys73(2), Cys44(1) Cys48(2), Cys70(1)-Cys7(2), as determined by electrospray mass spectrometry after tryptic digestion. Circular dichroism analysis showed identical folds of natural and recombinant Fel d 1. Furthermore, recombinant Fel d l reacted specifically with serum IgE, inducing expression of CD203c on basophils and lymphoproliferative responses in cat-allergic patients. The results show that the overall fold and immunological properties of the recombinant Fel d 1 are very similar to those of natural Fel d 1. Moreover, the recombinant Fel d 1 construct provides a tool for defining the three-dimensional structure of Fel d 1 and represents a reagent for diagnosis and allergen-specific immunotherapy of cat allergy. PMID- 12732624 TI - Molecular cloning and characterization of CYP719, a methylenedioxy bridge-forming enzyme that belongs to a novel P450 family, from cultured Coptis japonica cells. AB - Two cytochrome P450 (P450) cDNAs involved in the biosynthesis of berberine, an antimicrobial benzylisoquinoline alkaloid, were isolated from cultured Coptis japonica cells and characterized. A sequence analysis showed that one C. japonica P450 (designated CYP719) belonged to a novel P450 family. Further, heterologous expression in yeast confirmed that it had the same activity as a methylenedioxy bridge-forming enzyme (canadine synthase), which catalyzes the conversion of (S) tetrahydrocolumbamine ((S)-THC) to (S)-tetrahydroberberine ((S)-THB, (S) canadine). The other P450 (designated CYP80B2) showed high homology to California poppy (S)-N-methylcoclaurine-3'-hydroxylase (CYP80B1), which converts (S)-N methylcoclaurine to (S)-3'-hydroxy-N-methylcoclaurine. Recombinant CYP719 showed typical P450 properties as well as high substrate affinity and specificity for (S)-THC. (S)Scoulerine was not a substrate of CYP719, indicating that some other P450, e.g. (S)-cheilanthifoline synthase, is needed in (S)-stylopine biosynthesis. All of the berberine biosynthetic genes, including CYP719 and CYP80B2, were highly expressed in selected cultured C. japonica cells and moderately expressed in root, which suggests coordinated regulation of the expression of biosynthetic genes. PMID- 12732625 TI - Crystal structure of fungal lectin: six-bladed beta-propeller fold and novel fucose recognition mode for Aleuria aurantia lectin. AB - Aleuria aurantia lectin is a fungal protein composed of two identical 312-amino acid subunits that specifically recognizes fucosylated glycans. The crystal structure of the lectin complexed with fucose reveals that each monomer consists of a six-bladed beta-propeller fold and of a small antiparallel two-stranded beta sheet that plays a role in dimerization. Five fucose residues were located in binding pockets between the adjacent propeller blades. Due to repeats in the amino acid sequence, there are strong similarities between the sites. Oxygen atoms O-3, O-4, and O-5 of fucose are involved in hydrogen bonds with side chains of amino acids conserved in all repeats, whereas O-1 and O-2 interact with a large number of water molecules. The nonpolar face of each fucose residue is stacked against the aromatic ring of a Trp or Tyr amino acid, and the methyl group is located in a highly hydrophobic pocket. Depending on the precise binding site geometry, the alpha- or beta-anomer of the fucose ligand is observed bound in the crystal. Surface plasmon resonance experiments conducted on a series of oligosaccharides confirm the broad specificity of the lectin, with a slight preference for alphaFuc1-2Gal disaccharide. This multivalent carbohydrate recognition fold is a new prototype of lectins that is proposed to be involved in the host recognition strategy of several pathogenic organisms including not only the fungi Aspergillus but also the phytopathogenic bacterium Ralstonia solanacearum. PMID- 12732626 TI - Correlation of the sweetness of variants of the protein brazzein with patterns of hydrogen bonds detected by NMR spectroscopy. AB - In sequence-function investigations, approaches are needed for rapidly screening protein variants for possible changes in conformation. Recent NMR methods permit direct detection of hydrogen bonds through measurements of scalar couplings that traverse hydrogen bonds (trans-hydrogen bond couplings). We have applied this approach to screen a series of five single site mutants of the sweet protein brazzein with altered sweetness for possible changes in backbone hydrogen bonding with respect to wild-type. Long range, three-dimensional data correlating connectivities among backbone 1HN, 15N, and 13C' atoms were collected from the six brazzein proteins labeled uniformly with carbon-13 and nitrogen-15. In wild type brazzein, this approach identified 17 backbone hydrogen bonds. In the mutants, altered magnitudes of the couplings identified hydrogen bonds that were strengthened or weakened; missing couplings identified hydrogen bonds that were broken, and new couplings indicated the presence of new hydrogen bonds. Within the series of brazzein mutants investigated, a pattern was observed between sweetness and the integrity of particular hydrogen bonds. All three "sweet" variants exhibited the same pattern of hydrogen bonds, whereas all three "non sweet" variants lacked one hydrogen bond at the middle of the alpha-helix, where it is kinked, and one hydrogen bond in the middle of beta-strands II and III, where they are twisted. Two of the non-sweet variants lack the hydrogen bond connecting the N and C termini. These variants showed greater mobility in the N- and C-terminal regions than wild-type brazzein. PMID- 12732627 TI - Mice with targeted mutation of peroxiredoxin 6 develop normally but are susceptible to oxidative stress. AB - Reactive oxygen species, especially hydrogen peroxide, are important in cellular signal transduction. However, excessive amounts of these species damage tissues and cells by oxidizing virtually all important biomolecules. Peroxiredoxin 6 (PRDX6) (also called antioxidant protein 2, or AOP2) is a novel peroxiredoxin family member whose function in vivo is unknown. Through immunohistochemistry, we have determined that the PRDX6 protein was widely expressed in every tissue examined, most abundantly in epithelial cells. It was found in cytosol, but not in membranes, organelles, and nuclei fractions. Prdx6 mRNA was also expressed in every tissue examined. The widespread expression of Prdx6 suggested that its functions were quite important. To determine these functions, we generated Prdx6 targeted mutant (Prdx6-/-) mice, confirmed the gene disruption by Southern blots, PCR, RT-PCR, Western blots, and immunohistochemistry, and compared the effects of paraquat, hydrogen peroxide, and t-butyl hydroperoxide on Prdx6-/- and wild-type (Prdx6+/+) macrophages, and of paraquat on Prdx6-/- and Prdx6+/+ mice. Prdx6-/- macrophages had higher hydrogen peroxide levels, and lower survival rates; Prdx6 /- mice had significantly lower survival rates, more severe tissue damage, and higher protein oxidation levels. Additionally, there were no differences in the mRNA expression levels of other peroxiredoxins, glutathione peroxidases, catalase, superoxide dismutases, thioredoxins, and glutaredoxins between normal Prdx6-/- and Prdx6+/+ mice and those injected with paraquat. Our study provides in vivo evidence that PRDX6 is a unique non-redundant antioxidant that functions independently of other peroxiredoxins and antioxidant proteins. PMID- 12732628 TI - Mechanistic studies of human molybdopterin synthase reaction and characterization of mutants identified in group B patients of molybdenum cofactor deficiency. AB - Biosynthesis of the molybdenum cofactor involves the initial formation of precursor Z, its subsequent conversion to molybdopterin (MPT) by MPT synthase, and attachment of molybdenum to the dithiolene moiety of MPT. The sulfur used for the formation of the dithiolene group of MPT exists in the form of a thiocarboxylate group at the C terminus of the smaller subunit of MPT synthase. Human MPT synthase contains the MOCS2A and MOCS2B proteins that display homology to the Escherichia coli proteins MoaD and MoaE, respectively. MOCS2A and MOCS2B were purified after heterologous expression in E. coli, and the separately purified subunits readily assemble into a functional MPT synthase tetramer. The rate of conversion of precursor Z to MPT by the human enzyme is slower than that of the eubacterial homologue. To obtain insights into the molecular mechanism leading to human molybdenum cofactor deficiency, site-specific mutations identified in patients showing symptoms of molybdenum cofactor deficiency were generated. Characterization of a V7F substitution in MOCS2A, identified in a patient with an unusual mild form of the disease, showed that the mutation weakens the interaction between MOCS2A and MOCS2B, whereas a MOCS2B-E168K mutation identified in a severely affected patient attenuates binding of precursor Z. PMID- 12732629 TI - A novel RGD-independent cel adhesion pathway mediated by fibronectin-bound tissue transglutaminase rescues cells from anoikis. AB - Specific association of tissue transglutaminase (tTG) with matrix fibronectin (FN) results in the formation of an extracellular complex (tTG-FN) with distinct adhesive and pro-survival characteristics. tTG-FN supports RGD-independent cell adhesion of different cell types and the formation of distinctive RhoA-dependent focal adhesions following inhibition of integrin function by competitive RGD peptides and function blocking anti-integrin antibodies alpha5beta1. Association of tTG with its binding site on the 70-kDa amino-terminal FN fragment does not support this cell adhesion process, which seems to involve the entire FN molecule. RGD-independent cell adhesion to tTG-FN does not require transamidating activity, is mediated by the binding of tTG to cell-surface heparan sulfate chains, is dependent on the function of protein kinase Calpha, and leads to activation of the cell survival focal adhesion kinase. The tTG-FN complex can maintain cell viability of tTG-null mouse dermal fibroblasts when apoptosis is induced by inhibition of RGD-dependent adhesion (anoikis), suggesting an extracellular survival role for tTG. We propose a novel RGD-independent cell adhesion mechanism that promotes cell survival when the anti-apoptotic role mediated by RGD-dependent integrin function is reduced as in tissue injury, which is consistent with the externalization and binding of tTG to fibronectin following cell damage/stress. PMID- 12732630 TI - Genetic rescue of chondrodysplasia and the perinatal lethal effect of cartilage link protein deficiency. AB - The targeted disruption of cartilage link protein gene (Crtl1) in homozygous mice resulted in a severe chondrodysplasia and perinatal lethality. This raised the question of whether the abnormalities seen in Crtl1 null mice are all caused by the absence of link protein in cartilage or whether the deficiency of the protein in other tissues and organs contributed to the phenotype. To address this question we have generated transgenic mice overexpressing cartilage link protein under the control of a cartilage-specific promoter, and then these transgenic mice were used for a genetic rescue of abnormalities in Crtl1 null mice. While the overexpression of cartilage link protein resulted in no abnormal phenotype, the cartilage-specific transgene expression of link protein could completely prevent the perinatal mortality of link protein-deficient mice and, depending on the level of the link protein expression, rescue skeletal abnormalities. Although link protein was originally isolated from cartilage, we found and determined Crtl1 transcripts and corresponding proteins in every organ tested from mouse embryos to aging animals. We also identified three additional members of the link protein family, all co-localized with hyaluronic acid-binding proteoglycans in the mouse genome. The ubiquitous presence of link protein suggests a general and systemic function of link protein in the organization of extracellular matrix in a number of tissues, possibly interacting with other proteoglycans, such as versican, brevican, and neurocan. PMID- 12732631 TI - Transcriptional co-activators CREB-binding protein and p300 regulate chondrocyte specific gene expression via association with Sox9. AB - Chondrocytes are critical components for the precise patterning of a developing skeletal framework and articular joint formation. Sox9 is a key transcription factor that is essential for chondrocyte differentiation and chondrocyte-specific gene expressions; however, the precise transcriptional activation mechanism of Sox9 is not fully understood. Here we demonstrate that Sox9 utilizes a cAMP response element-binding protein (CREB)-binding protein (CBP)/p300 to exert its effects. Sox9 associates with CBP/p300 in the chondrosarcoma cell line SW1353 via its carboxyl termini activation domain in a cell type-specific manner. In promoter assays, CBP/p300 enhances Col2a1, which encodes cartilage-specific type II collagen gene promoter activity via Sox9. Chromatin immunoprecipitation shows that p300 is bound to the Col2a1 promoter region. Furthermore, the CBP/Sox9 complex disrupter peptide suppresses Col2a1 gene expression and chondrogenesis from mesenchymal stem cells. These data demonstrate that CBP and p300 function as co-activators of Sox9 for cartilage tissue-specific gene expression and chondrocyte differentiation. PMID- 12732632 TI - Enhanced immune presentation of a single-chain major histocompatibility complex class I molecule engineered to optimize linkage of a C-terminally extended peptide. AB - Major histocompatibility complex class I molecules can be expressed as single polypeptides wherein the antigenic peptide, beta2-microglobulin, and heavy chain are attached by flexible linkers. These molecules, single-chain trimers (SCTs), are remarkably stable at the cell surface compared with native (noncovalently attached) class I molecules. In this study, we used a structure-based approach to engineer an F pocket variant SCT of the murine class I molecule Kb that presents the SIINFEKL epitope of ovalbumin. Mutation of heavy chain residue Tyr84 (Y84A) in the SCT resulted in enhanced serological and cytolytic CD8 T cell recognition of the covalently linked peptide due to better accommodation of the linker extending from the C terminus of the peptide. These SCTs exhibit significant cell surface stability, which we hypothesize is rendered by their ability to continuously and efficiently rebind the covalently attached peptide. In addition, we demonstrate that SCT technology can be applied to tetramer construction using recombinant SCTs expressed in Escherichia coli. SCT-based tetramers could have applications for the enumeration of T and natural killer cells that recognize peptide.class I complexes prone to dissociation. PMID- 12732633 TI - Molecular and functional characterization of clathrin- and AP-2-binding determinants within a disordered domain of auxilin. AB - Uncoating of clathrin-coated vesicles requires the J-domain protein auxilin for targeting hsc70 to the clathrin coats and for stimulating the hsc70 ATPase activity. This results in the release of hsc70-complexed clathrin triskelia and concomitant dissociation of the coat. To understand the complex role of auxilin in uncoating and clathrin assembly in more detail, we analyzed the molecular organization of its clathrin-binding domain (amino acids 547-813). CD spectroscopy of auxilin fragments revealed that the clathrin-binding domain is almost completely disordered in solution. By systematic mapping using synthetic peptides and by site-directed mutagenesis, we identified short peptide sequences involved in clathrin heavy chain and AP-2 binding and evaluated their significance for the function of auxilin. Some of the binding determinants, including those containing sequences 674DPF and 636WDW, showed dual specificity for both clathrin and AP-2. In contrast, the two DLL motifs within the clathrin binding domain were exclusively involved in clathrin binding. Surprisingly, they interacted not only with the N-terminal domain of the heavy chain, but also with the distal domain. Moreover, both DLL peptides proved to be essential for clathrin assembly and uncoating. In addition, we found that the motif 726NWQ is required for efficient clathrin assembly activity. Auxilin shares a number of protein-protein interaction motifs with other endocytic proteins, including AP180. We demonstrate that AP180 and auxilin compete for binding to the alpha-ear domain of AP-2. Like AP180, auxilin also directly interacts with the ear domain of beta-adaptin. On the basis of our data, we propose a refined model for the uncoating mechanism of clathrin-coated vesicles. PMID- 12732634 TI - The oncoprotein Ski acts as an antagonist of transforming growth factor-beta signaling by suppressing Smad2 phosphorylation. AB - The phosphorylation of Smad2 and Smad3 by the transforming growth factor (TGF) beta-activated receptor kinases and their subsequent heterodimerization with Smad4 and translocation to the nucleus form the basis for a model how Smad proteins work to transmit TGF-beta signals. The transcriptional activity of Smad2 Smad4 or Smad3-Smad4 complexes can be limited by the corepressor Ski, which is believed to interact with Smad complexes on TGF-beta-responsive promoters and represses their ability to activate TGF-beta target genes by assembling on DNA a repressor complex containing histone deacetylase. Here we show that Ski can block TGF-beta signaling by interfering with the phosphorylation of Smad2 and Smad3 by the activated TGF-beta type I receptor. Furthermore, we demonstrate that overexpression of Ski induces the assembly of Smad2-Smad4 and Smad3-Smad4 complexes independent of TGF-beta signaling. The ability of Ski to engage Smad proteins in nonproductive complexes provides new insights into the molecular mechanism used by Ski for disabling TGF-beta signaling. PMID- 12732635 TI - IIGP1, an interferon-gamma-inducible 47-kDa GTPase of the mouse, showing cooperative enzymatic activity and GTP-dependent multimerization. AB - IIGP1 belongs to a well defined family of 47-kDa GTPases whose members are present at low resting levels in mouse cells but are strongly induced transcriptionally by interferons and are implicated in cell-autonomous resistance to intracellular pathogens. Recombinant IIGP1 was expressed in Escherichia coli and purified to homogeneity. Here we present a detailed biochemical characterization of IIGP1 using various biochemical and biophysical methods. IIGP1 binds to GTP and GDP with dissociation constants in the micromolar range with at least 10 times higher affinity for GDP than for GTP. IIGP1 hydrolyzes GTP to GDP, and the GTPase activity is concentration-dependent with a GTP turnover rate of 2 min-1 under saturating protein concentrations. Functional interaction between IIGP1 molecules is shown by nucleotide-dependent oligomerization in vitro. Both cooperative hydrolysis of GTP and GTP-dependent oligomerization are blocked in a mutant form of IIGP1 modified at the C terminus. IIGP1 shares micromolar nucleotide affinities and oligomerization-dependent hydrolytic activity with the 67-kDa GTPase hGBP1 (induced by type I and type II interferons), with the antiviral Mx proteins (interferon type I-induced) and with the paradigm of the self-activating large GTPases, the dynamins, with which Mx proteins show homology. The higher relative affinity for GDP and the relatively low GTPase activity distinguish IIGP1, but this study clearly adds IIGP1 and thus the p47 GTPases to the small group of cooperative GTPase families that appear to characterize the development of intracellular resistance during the interferon response to infection. The present analysis provides essential parameters to understand the molecular mechanism by which IIGP1 participates in this complex resistance program. PMID- 12732637 TI - Structures of liganded and unliganded RsrI N6-adenine DNA methyltransferase: a distinct orientation for active cofactor binding. AB - The structures of RsrI DNA methyltransferase (M.RsrI) bound to the substrate S adenosyl-l-methionine (AdoMet), the product S-adenosyl-l-homocysteine (AdoHcy), the inhibitor sinefungin, as well as a mutant apo-enzyme have been determined by x-ray crystallography. Two distinct binding configurations were observed for the three ligands. The substrate AdoMet adopts a bent shape that directs the activated methyl group toward the active site near the catalytic DPPY motif. The product AdoHcy and the competitive inhibitor sinefungin bind with a straight conformation in which the amino acid moiety occupies a position near the activated methyl group in the AdoMet complex. Analysis of ligand binding in comparison with other DNA methyltransferases reveals a small, common subset of available conformations for the ligand. The structures of M.RsrI with the non substrate ligands contained a bound chloride ion in the AdoMet carboxylate binding pocket, explaining its inhibition by chloride salts. The L72P mutant of M.RsrI is the first DNA methyltransferase structure without bound ligand. With respect to the wild-type protein, it had a larger ligand-binding pocket and displayed movement of a loop (223-227) that is responsible for binding the ligand, which may account for the weaker affinity of the L72P mutant for AdoMet. These studies show the subtle changes in the tight specific interactions of substrate, product, and an inhibitor with M.RsrI and help explain how each displays its unique effect on the activity of the enzyme. PMID- 12732636 TI - Caveolin-1 contributes to assembly of store-operated Ca2+ influx channels by regulating plasma membrane localization of TRPC1. AB - TRPC1, a component of store-operated Ca2+ entry (SOCE) channels, is assembled in a complex with caveolin-1 (Cav1) and key Ca2+ signaling proteins. This study examines the role of Cav1 in the function of TRPC1. TRPC1 and Cav1 were colocalized in the plasma membrane region of human submandibular gland and Madin Darby canine kidney cells. Full-length Cav1 bound to both the N and C termini of TRPC1. Amino acids 271-349, which includes a Cav1 binding motif (amino acids 322 349), was identified as the Cav1 binding domain in the TRPC1 N terminus. Deletion of amino acids 271-349 or 322-349 prevented plasma membrane localization of TRPC1. Importantly, TRPC1Delta271-349 induced a dominant suppression of SOCE and was associated with wild-type TRPC1. Although the role of the C-terminal Cav1 binding domain is not known, its deletion did not affect localization of TRPC1 (Singh, B. B., Liu, X., and Ambudkar, I. S. (2000) J. Biol. Chem. 275, 36483 36486). Further, expression of a truncated Cav1 (Cav1Delta51-169), but not full length Cav1, similarly disrupted plasma membrane localization of endogenously and exogenously expressed TRPC1 in human submandibular gland and Madin-Darby canine kidney cells. Cav1Delta51-169 also suppressed thapsigarginand carbachol stimulated Ca2+ influx and increased the detergent solubility of TRPC1, although plasma membrane lipid raft domains were not disrupted. These data demonstrate that plasma membrane localization of TRPC1 depends on an interaction between its N terminus and Cav1. Thus, our data suggest that Cav1 has an important role in the assembly of SOCE channel(s). PMID- 12732638 TI - Sequential interaction of actin-related proteins 2 and 3 (Arp2/3) complex with neural Wiscott-Aldrich syndrome protein (N-WASP) and cortactin during branched actin filament network formation. AB - The WASP and cortactin families constitute two distinct classes of Arp2/3 modulators in mammalian cells. Physical and functional interactions among the Arp2/3 complex, VCA (a functional domain of N-WASP), and cortactin were examined under conditions that were with or without actin polymerization. In the absence of actin, cortactin binds significantly weaker to the Arp2/3 complex than VCA. At concentrations of VCA 20-fold lower than cortactin, the association of cortactin with the Arp2/3 complex was nearly abolished. Analysis of the cells infected with Shigella demonstrated that N-WASP located at the tip of the bacterium, whereas cortactin accumulated in the comet tail. Interestingly, cortactin promotes Arp2/3 complex-mediated actin polymerization and actin branching in the presence of VCA at a saturating concentration, and cortactin acquired 20 nm affinity for the Arp2/3 complex during actin polymerization. The interaction of VCA with the Arp2/3 complex was reduced in the presence of both cortactin and actin. Moreover, VCA reduced its affinity for Arp2/3 complex at branching sites that were stabilized by phalloidin. These data imply a novel mechanism for the de novo assembly of a branched actin network that involves a coordinated sequential interaction of N-WASP and cortactin with the Arp2/3 complex. PMID- 12732640 TI - p116Rip is a novel filamentous actin-binding protein. AB - p116Rip is a ubiquitously expressed protein that was originally identified as a putative binding partner of RhoA in a yeast two-hybrid screen. Overexpression of p116Rip in neuroblastoma cells inhibits RhoA-mediated cell contraction induced by lysophosphatidic acid (LPA); so far, however, the function of p116Rip is unknown. Here we report that p116Rip localizes to filamentous actin (F-actin)-rich structures, including stress fibers and cortical microfilaments, in both serum deprived and LPA-stimulated cells, with the N terminus (residues 1-382) dictating cytoskeletal localization. In addition, p116Rip is found in the nucleus. Direct interaction or colocalization with RhoA was not detected. We find that p116Rip binds tightly to F-actin (Kd approximately 0.5 microm) via its N-terminal region, while immunoprecipitation assays show that p116Rip is complexed to both F-actin and myosin-II. Purified p116Rip and the F-actin-binding region can bundle F-actin in vitro, as shown by electron microscopy. When overexpressed in NIH3T3 cells, p116Rip disrupts stress fibers and promotes formation of dendrite-like extensions through its N-terminal actin-binding domain; furthermore, overexpressed p116Rip inhibits growth factor-induced lamellipodia formation. Our results indicate that p116Rip is an F-actin-binding protein with in vitro bundling activity and in vivo capability of disassembling the actomyosin-based cytoskeleton. PMID- 12732639 TI - Functional defects in six ryanodine receptor isoform-1 (RyR1) mutations associated with malignant hyperthermia and their impact on skeletal excitation contraction coupling. AB - Malignant hyperthermia (MH) is a potentially fatal pharmacogenetic disorder of skeletal muscle that segregates with >60 mutations within the MHS-1 locus on chromosome 19 coding for ryanodine receptor type 1 (RyR1). Although some MHRyR1s have been shown to enhance sensitivity to caffeine and halothane when expressed in non-muscle cells, their influence on EC coupling can only be studied in skeletal myotubes. We therefore expressed WTRyR1, six of the most common human MHRyR1s (R163C, G341R, R614C, R2163C, V2168M, and R2458H), and a newly identified C-terminal mutation (T4826I) in dyspedic myotubes to study their functional defects and how they influence EC coupling. Myotubes expressing any MHRyR1 were significantly more sensitive to stimulation by caffeine and 4-CmC than those expressing WTRyR1. The hypersensitivity of MH myotubes extended to K+ depolarization. MH myotubes responded to direct channel activators with maximum Ca2+ amplitudes consistently smaller than WT myotubes, whereas the amplitude of their responses to depolarization were consistently larger than WT myotubes. The magnitudes of responses attainable from myotubes expressing MHRyR1s are therefore related to the nature of the stimulus rather than size of the Ca2+ store. The functional changes of MHRyR1s were directly analyzed using [3H]ryanodine binding analysis of isolated myotube membranes. Although none of the MHRyR1s examined significantly altered EC50 for Ca2+ activation, many failed to be completely inhibited by a low Ca2+ (2000 atoms of iron in 2-4-nm cores structurally similar to ferritin iron cores. Here we show that mYfh1p assembly is driven by two sequential iron oxidation reactions: A ferroxidase reaction catalyzed by mYfh1p induces the first assembly step (alpha --> alpha3), followed by a slower autoxidation reaction that promotes the assembly of higher order oligomers yielding alpha48. Depending on the ionic environment, stepwise assembly is associated with accumulation of 50-75 Fe(II)/subunit. Initially, this Fe(II) is loosely bound to mYfh1p and can be readily mobilized by chelators or made available to the mitochondrial enzyme ferrochelatase to synthesize heme. Transfer of mYfh1p-bound Fe(II) to ferrochelatase occurs in the presence of citrate, a physiologic ferrous iron chelator, suggesting that the transfer involves an intermolecular interaction. If mYfh1p-bound Fe(II) is not transferred to a ligand, iron oxidation, and mineralization proceed to completion, Fe(III) becomes progressively less accessible, and a stable iron-protein complex is formed. Iron oxidation-driven stepwise assembly is a novel mechanism by which yeast frataxin can function as an iron chaperone or an iron store. PMID- 12732650 TI - Lactococcus lactis dihydroorotate dehydrogenase A mutants reveal important facets of the enzymatic function. AB - Dihydroorotate dehydrogenases (DHODs) are flavoenzymes catalyzing the oxidation of (S)-dihydroorotate to orotate in the biosynthesis of UMP, the precursor of all other pyrimidine nucleotides. On the basis of sequence, DHODs can be divided into two classes, class 1, further divided in subclasses 1A and 1B, and class 2. This division corresponds to differences in cellular location and the nature of the electron acceptor. Herein we report a study of Lactococcus lactis DHODA, a representative of the class 1A enzymes. Based on the DHODA structure we selected seven residues that are highly conserved between both main classes of DHODs as well as three residues representing surface charges close to the active site for site-directed mutagenesis. The availability of both kinetic and structural data on the mutant enzymes allowed us to define the roles individual structural segments play in catalysis. We have also structurally proven the presence of an open active site loop in DHODA and obtained information about the interactions that control movements of loops around the active site. Furthermore, in one mutant structure we observed differences between the two monomers of the dimer, confirming an apparent asymmetry between the two substrate binding sites that was indicated by the kinetic results. PMID- 12732652 TI - SOX8 is expressed during testis differentiation in mice and synergizes with SF1 to activate the Amh promoter in vitro. AB - Sox8 is a member of the Sox family of developmental transcription factor genes and is closely related to Sox9, a key gene in the testis determination pathway in mammals. Like Sox9, Sox8 is expressed in the developing mouse testis around the time of sex determination, suggesting that it might play a role in regulating the expression of testis-specific genes. An early step in male sex differentiation is the expression of anti-Mullerian hormone (AMH) in Sertoli cells. Expression of the Amh gene during sex differentiation requires the interaction of several transcription factors, including SF1, SOX9, GATA4, WT1, and DAX1. Here we show that SOX8 may also be involved in regulating the expression of Amh. Expression of Sox8 begins just prior to that of Amh at 12 days post coitum (dpc) in mouse testes and continues beyond 16 dpc in Sertoli cells. In vitro assays showed that SOX8 binds specifically to SOX binding sites within the Amh minimal promoter and, like SOX9, acts synergistically with SF1 through direct protein-protein interaction to enhance Amh expression, albeit at lower levels compared with SOX9. SOX8 and SOX9 appear to have arisen from a common ancestral gene and may have retained some common functions during sexual development. Our data provide the first evidence that SOX8 may partially compensate for the reduced SOX9 activity in campomelic dysplasia and substitute for Sox9 where Sox9 is either not expressed or expressed too late to be involved in sex determination or regulation of Amh expression. PMID- 12732651 TI - Integrating structure, bioinformatics, and enzymology to discover function: BioH, a new carboxylesterase from Escherichia coli. AB - Structural proteomics projects are generating three-dimensional structures of novel, uncharacterized proteins at an increasing rate. However, structure alone is often insufficient to deduce the specific biochemical function of a protein. Here we determined the function for a protein using a strategy that integrates structural and bioinformatics data with parallel experimental screening for enzymatic activity. BioH is involved in biotin biosynthesis in Escherichia coli and had no previously known biochemical function. The crystal structure of BioH was determined at 1.7 A resolution. An automated procedure was used to compare the structure of BioH with structural templates from a variety of different enzyme active sites. This screen identified a catalytic triad (Ser82, His235, and Asp207) with a configuration similar to that of the catalytic triad of hydrolases. Analysis of BioH with a panel of hydrolase assays revealed a carboxylesterase activity with a preference for short acyl chain substrates. The combined use of structural bioinformatics with experimental screens for detecting enzyme activity could greatly enhance the rate at which function is determined from structure. PMID- 12732653 TI - Optic neuritis, a new variant of experimental encephalomyelitis, a durable model for all seasons, now in its seventieth year. PMID- 12732654 TI - Myelin oligodendrocyte glycoprotein-specific T cell receptor transgenic mice develop spontaneous autoimmune optic neuritis. AB - Multiple sclerosis (MS) is considered to be an autoimmune disease of the central nervous system (CNS) that in many patients first presents clinically as optic neuritis. The relationship of optic neuritis to MS is not well understood. We have generated novel T cell receptor (TCR) transgenic mice specific for myelin oligodendrocyte glycoprotein (MOG). MOG-specific transgenic T cells are not deleted nor tolerized and are functionally competent. A large proportion (>30%) of MOG-specific TCR transgenic mice spontaneously develop isolated optic neuritis without any clinical nor histological evidence of experimental autoimmune encephalomyelitis (EAE). Optic neuritis without EAE could also be induced in these mice by sensitization with suboptimal doses of MOG. The predilection of these mice to develop optic neuritis is associated with higher expression of MOG in the optic nerve than in the spinal cord. These results demonstrate that clinical manifestations of CNS autoimmune disease will vary depending on the identity of the target autoantigen and that MOG-specific T cell responses are involved in the genesis of isolated optic neuritis. PMID- 12732655 TI - Enhanced antitumor immunity in mice deficient in CD69. AB - We investigated the in vivo role of CD69 by analyzing the susceptibility of CD69 /- mice to tumors. CD69-/- mice challenged with MHC class I- tumors (RMA-S and RM 1) showed greatly reduced tumor growth and prolonged survival compared with wild type (WT) mice. The enhanced anti-tumor response was NK cell and T lymphocyte mediated, and was due, at least in part, to an increase in local lymphocytes. Resistance of CD69-/- mice to MHC class I- tumor growth was also associated with increased production of the chemokine MCP-1, diminished TGF-beta production, and decreased lymphocyte apoptosis. Moreover, the in vivo blockade of TGF-beta in WT mice resulted in enhanced anti-tumor response. In addition, CD69 engagement induced NK and T cell production of TGF-beta, directly linking CD69 signaling to TGF-beta regulation. Furthermore, anti-CD69 antibody treatment in WT mice induced a specific down-regulation in CD69 expression that resulted in augmented anti tumor response. These data unmask a novel role for CD69 as a negative regulator of anti-tumor responses and show the possibility of a novel approach for the therapy of tumors. PMID- 12732656 TI - Signal 3 determines tolerance versus full activation of naive CD8 T cells: dissociating proliferation and development of effector function. AB - Activation of naive CD8 T cells to undergo clonal expansion and develop effector function requires three signals: (a) Ag, (b) costimulation, and (c) IL-12 or adjuvant. The requirement for the third signal to stimulate Ag-dependent proliferation is variable, making the greatest contribution when Ag levels are low. At high Ag levels, extensive proliferation can occur in vitro or in vivo in the absence of a third signal. However, despite having undergone the same number of divisions, cells that expand in the absence of a third signal fail to develop cytolytic effector function. Thus, proliferation and development of cytolytic function can be fully uncoupled. Furthermore, these cells are rendered functionally tolerant in vivo, in that subsequent restimulation with a potent stimulus results in limited clonal expansion, impaired IFN-gamma production, and no cytolytic function. Thus, the presence or absence of the third signal appears to be a critical variable in determining whether stimulation by Ag results in tolerance versus development of effector function and establishment of a responsive memory population. PMID- 12732657 TI - Ectopic LT alpha beta directs lymphoid organ neogenesis with concomitant expression of peripheral node addressin and a HEV-restricted sulfotransferase. AB - Lymph node (LN) function depends on T and B cell compartmentalization, antigen presenting cells, and high endothelial venules (HEVs) expressing mucosal addressin cell adhesion molecule (MAdCAM-1) and peripheral node addressin (PNAd), ligands for naive cell entrance into LNs. Luminal PNAd expression requires a HEV restricted sulfotransferase (HEC-6ST). To investigate LT alpha beta's activities in lymphoid organogenesis, mice simultaneously expressing LT alpha and LT beta under rat insulin promoter II (RIP) control were compared with RIPLT alpha mice in a model of lymphoid neogenesis and with LT beta-/- mice. RIPLT alpha beta pancreata exhibited massive intra-islet mononuclear infiltrates that differed from the more sparse peri-islet cell accumulations in RIPLT alpha pancreata: separation into T and B cell areas was more distinct with prominent FDC networks, expression of lymphoid chemokines (CCL21, CCL19, and CXCL13) was more intense, and L-selectin+ cells were more frequent. In contrast to the predominant abluminal PNAd pattern of HEV in LT beta-/- MLN and RIPLT alpha pancreatic infiltrates, PNAd was expressed at the luminal and abluminal aspects of HEV in wild-type LN and in RIPLT alpha beta pancreata, coincident with HEC-6ST. These data highlight distinct roles of LT alpha and LT alpha beta in lymphoid organogenesis supporting the notion that HEC-6ST-dependent luminal PNAd is under regulation by LT alpha beta. PMID- 12732658 TI - Constitutive expression of AID leads to tumorigenesis. AB - Genome stability is regulated by the balance between efficiencies of the repair machinery and genetic alterations such as mutations and chromosomal rearrangements. It has been postulated that deregulation of class switch recombination (CSR) and somatic hypermutation (SHM), which modify the immunoglobulin (Ig) genes in activated B cells, may be responsible for aberrant chromosomal translocations and mutations of non-Ig genes that lead to lymphocyte malignancy. However, the molecular basis for these genetic instabilities is not clearly understood. Activation-induced cytidine deaminase (AID) is shown to be essential and sufficient to induce both CSR and SHM in artificial substrates in fibroblasts as well as B cells. Here we show that constitutive and ubiquitous expression of AID in transgenic mice caused both T cell lymphomas and dysgenetic lesions of epithelium of respiratory bronchioles (micro-adenomas) in all individual mice. Point mutations, but not translocations, were massively introduced in expressed T cell receptor (TCR) and c-myc genes in T lymphoma cells. The results indicate that AID can mutate non-Ig genes including oncogenes, implying that aberrant AID expression could be a cause of human malignancy. PMID- 12732659 TI - A role for complement in feedback enhancement of antibody responses by IgG3. AB - IgG1, IgG2a, and IgG2b, passively administered with soluble Ags, enhance specific Ab responses. The effect of IgG3 in this type of feedback regulation has not been studied previously. We immunized mice with trinitrophenyl (TNP)-coupled carrier proteins (bovine serum albumin [BSA] or ovalbumin [OVA]) alone or complexed to monoclonal TNP-specific IgG3. The carrier-specific Ab responses were enhanced by several hundred-fold by IgG3. Enhancement was significantly impaired in mice depleted of complement factor C3 and in mice lacking complement receptors 1 and 2 (Cr2-/-). In contrast, mice lacking the common Fc-receptor gamma chain (FcR gamma -/-), resulting in reduced expression of Fc gamma RI and lack of Fc gamma RIII, and mice lacking Fc gamma RIIB (Fc gamma RIIB-/-), responded equally well to immunization with IgG3-complexed Ag as wild-type controls. These findings demonstrate that IgG3 can induce feedback enhancement and that IgG3, in analogy with IgM, uses the complement system for this function. PMID- 12732661 TI - Cooperating mechanisms of CXCR5 and CCR7 in development and organization of secondary lymphoid organs. AB - Homeostatic chemokines participate in the development of secondary lymphoid organs and later on in the functional organization of these tissues. The development of lymph nodes (LNs) and Peyer's patches depends on the recruitment of CD3- CD4+ interleukin (IL)-7R alpha hi cells to sites of future organ development. CD3- CD4+ IL-7R alpha hi cells express the chemokine receptor CXCR5 and might be attracted by its ligand CXCL13, which is secreted by mesenchymal cells. Mesenchymal cells also secrete CCL19, a ligand for CCR7, yet it is not clear whether CCR7 and CCL19 are important for secondary lymphoid organ development. Analyzing CXCR5-/- CCR7-/- double deficient mice we now show that these mice lack all examined peripheral LNs suggesting a profound role for both receptors in secondary lymphoid organ development. We demonstrate that CD3- CD4+ IL-7R alpha hi cells express CXCR5 as well as CCR7 indicating that both receptors cooperate during an early step of secondary lymphoid organ development. Furthermore, CXCR5-/- CCR7-/- mice display a severely disturbed architecture of mesenteric LN and spleen. Due to an impaired migration of B cells into the white pulp, CXCR5-/- CCR7-/- mice fail to develop B cell follicles but show small clusters of unorganized lymphocytes in the spleen. These data demonstrate a cooperative function of CXCR5 and CCR7 in lymphoid organ organogenesis and organization. PMID- 12732660 TI - Overlapping roles of CXCL13, interleukin 7 receptor alpha, and CCR7 ligands in lymph node development. AB - Lymphoid tissue development is associated with local accumulation of CD4+ CD3- IL 7R alpha hi hematopoietic cells that deliver lymphotoxin (LT)alpha 1 beta 2 signals to resident stromal cells. Previous studies have established an important role for CXCL13 (BLC) in the development of Peyer's patches (PP) and some peripheral lymph nodes (LNs), but the chemokine requirements for several LN types, including mesenteric LNs, remain undefined. Using CXCL13-/- mice that additionally carry the paucity of LN T cell mutation (plt/plt), we discovered that CCR7 ligands function in peripheral LN development. We also tested for a genetic interaction during LN development between CXCL13 and a cytokine receptor required in PP development, IL-7R alpha. Mice deficient for both CXCL13 and IL-7R alpha displayed a striking absence of LNs, including mesenteric LNs. These data extend the role of CXCL13 to the development of all LNs and establish a previously unappreciated role for IL-7R alpha in this process. Both circulating and LN CD4+ CD3- IL-7R alpha hi cells are shown to express LT alpha 1 beta 2 in an IL-7R alpha-dependent manner. Furthermore, CXCL13 was found to be sufficient to mediate CD4+ CD3- IL-7R alpha hi cell recruitment in vivo to an ectopic site. These findings indicate that CXCL13 and CCR7 ligands promote accumulation of CD4+ CD3- IL-7R alpha hi cells, delivering IL-7R alpha-dependent LT alpha 1 beta 2 signals critical for LN development. PMID- 12732662 TI - B cell-specific transgenic expression of Bcl2 rescues early B lymphopoiesis but not B cell responses in BOB.1/OBF.1-deficient mice. AB - Mice deficient for the transcriptional coactivator BOB.1/OBF.1 show several defects in B cell differentiation. Numbers of immature transitional B cells in the bone marrow are reduced and fewer B cells reach the periphery. Furthermore, germinal center B cells are absent and marginal zone (MZ) B lymphocytes are markedly reduced. Increased levels of B cell apoptosis in these mice prompted us to analyze expression and function of antiapoptotic proteins. Bcl2 expression is strongly reduced in BOB.1/OBF.1-deficient pre-B cells. When BOB.1/OBF.1-deficient mice were crossed with Bcl2-transgenic mice, B cell development in the bone marrow and numbers of B cells in peripheral lymphoid organs were normalized. However, neither germinal center B cells nor MZ B cells were rescued. Additionally, Bcl2 did not rescue the defects in signaling and affinity maturation found in BOB.1/OBF.1-deficient mice. Interestingly, Bcl2-transgenic mice by themselves show an MZ B cell defect. Virtually no functional MZ B cells were detected in these mice. In contrast, mice deficient for Bcl2 show a relative increase in MZ B cell numbers, indicating a previously undetected function of Bcl2 for this B cell compartment. PMID- 12732665 TI - The art of publishing methods. PMID- 12732663 TI - Deficiencies of GM-CSF and interferon gamma link inflammation and cancer. AB - Chronic inflammation contributes to carcinogenesis, but the underlying mechanisms are poorly understood. We report that aged granulocyte-macrophage colony stimulating factor (GM-CSF)-deficient mice develop a systemic lupus erythematosis (SLE)-like disorder associated with the impaired phagocytosis of apoptotic cells. Concurrent deficiency of interferon (IFN)-gamma attenuates the SLE, but promotes the formation of diverse hematologic and solid neoplasms within a background of persistent infection and inflammation. Whereas activated B cells show a resistance to fas-induced apoptosis, antimicrobial therapy prevents lymphomagenesis and solid tumor development. These findings demonstrate that the interplay of infectious agents with cytokine-mediated regulation of immune homeostasis is a critical determinant of cancer susceptibility. PMID- 12732664 TI - Regulation of Fyn through translocation of activated Lck into lipid rafts. AB - Whether or how the activation of Lck and Fyn during T cell receptor (TCR) signaling is coordinated, and their delivery of function integrated, is unknown. Here we show that lipid rafts function to segregate Lck and Fyn in T cells before activation. Coaggregation of TCR and CD4 leads to Lck activation within seconds outside lipid rafts, followed by its translocation into lipid rafts and the activation of colocalized Fyn. Genetic evidence demonstrates that Fyn activation is strictly dependent on receptor-induced translocation of Lck. These results characterize the interdependence of Lck and Fyn function and establish the spatial and temporal distinctions of their roles in the cellular activation process. PMID- 12732667 TI - A truly simultaneous combination of functional transcranial Doppler sonography and H(2)(15)O PET adds fundamental new information on differences in cognitive activation between schizophrenics and healthy control subjects. AB - Working memory deficits are a cardinal feature of the pathophysiology of schizophrenia. Lesion studies and functional blood flow-dependent imaging methods with coarse temporal resolution, such as PET and functional MRI (fMRI), tend to paint a fairly static picture of the cortical regions involved. In contrast, functional transcranial Doppler sonography (fTCD) provides a high temporal resolution. Truly simultaneous fTCD-fMRI is not yet possible for technical reasons, but H(2)(15)O PET and fTCD can be used really simultaneously. However, this combination has not yet been used for cognitive activations in schizophrenia. We therefore investigated the extent to which there are both spatial (PET) and temporal changes (fTCD) in the activation patterns of schizophrenic patients. METHODS: Eleven clinically stable chronic schizophrenic, right-handed patients and 10 healthy, right-handed control subjects, matched for age, sex, education, and intelligence quotient, participated in the study. We selected stable chronic schizophrenic patients who could perform a working memory task (N-back task) as well as healthy volunteers to exclude the possibility of imaged artifacts due to poor performance. All subjects were examined with a truly simultaneous fTCD-H(2)(15)O PET combination under cognitive activation. RESULTS: Schizophrenic patients activate a significantly larger cortical volume for adequate task performance (P < 0.05), but with a significantly lower blood flow increase in this volume (P < 0.01), than do control subjects. Furthermore, they cannot significantly increase blood flow velocity during the time course of cognitive activation as control subjects do. There were only significant correlations between neuropsychologic performance and imaging parameters (fTCD changes, PET blood flow changes) in control subjects (all r >or= /0.65/; P < 0.05), but no significant correlations in schizophrenics (all r < /0.3/; P > 0.4). CONCLUSION: We demonstrated that schizophrenic patients exhibit qualitative differences in the spatial and temporal resolution of cognitive processing. All facts could be interpreted as a sign of alternative, less efficient problem solving strategies in schizophrenia that lead to the working memory deficits observed during the further course of this disease. Truly simultaneous fTCD-PET can be used in neuroscience to add fundamental new information on spatial and temporal cognitive activation behavior to understand the true physiologic nature of the disease-specific differences of mental illnesses that are seen as disorders of the mind arising in the brain. PMID- 12732666 TI - Quantification of dopamine transporter by 123I-PE2I SPECT and the noninvasive Logan graphical method in Parkinson's disease. AB - (E)-N-(3-iodoprop-2-enyl)-2beta-carbomethoxy-3beta-(4'-methyl-phenyl) nortropane (PE2I), a cocaine analog, is a new, highly specific tracer for imaging dopamine transporter labeled with (123)I for in vivo SPECT. Its reversible binding on dopamine transporter and its rapid kinetics allow quantification of its binding potential according to a 3-compartment model. For quantification of distribution volume of reversible tracer, Logan developed a noninvasive and graphical method that allows accurate estimation of binding potential. In this study, we performed (123)I-PE2I SPECT on healthy volunteers and patients with Parkinson's disease (PD) to validate the Logan graphical method for quantification of (123)I-PE2I binding and to analyze the relationship between (123)I-PE2I SPECT and clinical features of this frequent degenerative disease. METHODS: Eight PD patients (3 women, 5 men; mean age, 64 +/- 7.9 y; disease duration range, 1-8 y, Hoehn and Yahr stage range, 1-2.5) and 8 age-matched healthy volunteers (4 women, 4 men; mean age, 61.5 +/- 9.5 y) were included in 2 centers and studied with SPECT. Four sequential SPECT imaging sessions of 15-min duration were performed from 5 to 65 min after bolus injection of 140 +/- 30 MBq of (123)I-PE2I. RESULTS: The kinetics of PE2I in healthy volunteers and PD patients were rapid, and the Logan graphical method allowed quantification of distribution volume ratio (DVR) in the caudate nucleus and putamen. (123)I-PE2I striatal specific binding was significantly reduced in PD patients, compared with healthy volunteers, in the caudate and putamen. The decrease of DVR in the putamen was significantly and inversely correlated to disease duration and Hoehn and Yahr stage. In asymmetric PD patients, (123)I-PE2I uptake was significantly more reduced in the putamen contralateral to the side with predominant clinical symptoms. However, (123)I PE2I uptake was also significantly reduced in the ipsilateral putamen, compared with that in healthy volunteers, suggesting that (123)I-PE2I SPECT can detect nigrostriatal degeneration before the appearance of clinical symptoms. CONCLUSION: Our data indicate that the Logan graphical method is accurate for noninvasive quantification of PE2I and that (123)I-PE2I SPECT is a useful quantitative method for accurate estimation of nigrostriatal dopaminergic nerve terminal degeneration. The close relationships between SPECT findings and clinical data suggest that this method is useful for objectively following the progression of PD and for assessing the effect of potential neuroprotective treatments. Finally, our findings suggest that (123)I-PE2I SPECT can be used for preclinical and early diagnosis of PD. PMID- 12732668 TI - Insulin stimulates liver glucose uptake in humans: an 18F-FDG PET Study. AB - The liver is vital for the regulation of glucose metabolism, but inaccessibility of the organ for direct assessments has limited the study of its metabolic role in vivo. METHODS: The effect of insulin and insulin sensitivity (IS) on hepatic glucose uptake was investigated using PET, (18)F-FDG, and graphical analysis and 3-compartment modeling in humans. We studied 16 healthy sedentary men (normal IS), 8 athletes (high IS), and 7 patients with coronary artery disease (low IS) either during fasting (n = 14) or during euglycemic hyperinsulinemia (n = 24). RESULTS: Whole-body insulin-mediated glucose uptake was 35 +/- 7 micro mol/min/kg for normal-IS subjects, 65 +/- 8 micro mol/min/kg for high-IS subjects (P < 0.05 vs. normal IS), and 24 +/- 3 micro mol/min/kg for low-IS subjects (P < 0.05 vs. normal IS and high IS). Hyperinsulinemia enhanced hepatic glucose influx (2.3 +/- 0.9 vs. 1.5 +/- 0.7 micro mol x min(-1) x 100 mL(-1), P < 0.05) and phosphorylation rates (0.55 +/- 0.24 vs. 0.36 +/- 0.19 min(-1) x 10(-2), P < 0.05) similarly in insulin-sensitive and -resistant subjects. During hyperinsulinemia, however, the glucose phosphorylation-to-dephosphorylation ratio was significantly lower in the low-IS group than in normal-IS subjects (P < 0.05) or high-IS subjects (P < 0.01); correspondingly, whole-body insulin-mediated glucose disposal was directly related to this ratio (r = 0.45; P < 0.05). Furthermore, glucose influx rates were inversely correlated with fasting plasma free fatty acids (P < 0.05). Both compartmental modeling and the graphical approach accurately described the data, though the latter yielded slightly lower estimates of glucose influx rates during fasting. CONCLUSION: Our study provided evidence that physiologic hyperinsulinemia enhances hepatic glucose uptake and that IS is related to the glucose phosphorylation-to-dephosphorylation balance in the liver. Graphical analysis and modeling proved to be applicable and complementary tools for the investigation of glucose metabolism in the liver. PMID- 12732669 TI - Evaluation of 18F-FDG PET in patients with advanced, metastatic, or recurrent gastric cancer. AB - PET with 18F-FDG has been widely used in oncology, but its application for stomach neoplasms has been limited. The aim of this study was to evaluate the visual diagnostic accuracy of (18)F-FDG PET for advanced, metastatic, or recurrent gastric cancer and to generate semiquantitative values for lesions. METHODS: 18F-FDG PET scans were obtained on 42 patients (29 men, 13 women; age, 27-78 y; median age, 63 y): 20 patients with a PT931/04 scanner and 22 patients with a SET2400W scanner. The PT931/04 has a spatial resolution of 6.0 mm at full width at half maximum (FWHM) and covers 15 cm above and below the targeted lesion, and the SET2400W has a spatial resolution of 3.9 mm at FWHM and images the entire body. All PET images were interpreted visually, and tracer uptakes were quantitated as standardized uptake values (SUVs) on SET2400W images. RESULTS: The sensitivity, specificity, and accuracy as a whole were as follows: 71%, 74%, and 73%, respectively, with the SET2400W scanner and 47%, 79%, and 62%, respectively, with the PT931/04 scanner. Values were high for primary lesions, liver, lymph node, and lung metastases, but were low for bone metastases, ascites, peritonitis, and pleuritis carcinomatoses. SUVs were 8.9 +/- 4.2 (primary lesions, 19 patients/19 lesions), 6.5 +/- 2.2 (liver, 9/55), 6.1 +/- 2.5 (lymph nodes, 14/38), 6.5 +/- 1.8 (abdominal wall, 4/7), 3.9 +/- 2.0 (bone, 3/27), and 4.7 +/- 2.6 (lung, 2/3). Comparing SUVs and histologic findings for 17 untreated patients, values for well-differentiated and moderately differentiated adenocarcinomas versus poorly differentiated adenocarcinomas and signet ring cell carcinomas were 13.2 +/- 6.3 (4/4) versus 7.7 +/- 2.6 (13/13) (P < 0.05) for the primary lesions, 7.0 +/- 2.4 (5/39) versus 5.6 +/- 2.8 (2/2) for the liver, and 5.5 +/- 1.9 (9/28) versus 8.8 +/- 3.3 (3/8) (P < 0.05) for the lymph nodes. CONCLUSION: Our results indicate that 18F-FDG PET is a useful diagnostic modality for advanced, metastatic, or recurrent gastric cancer but not for detecting bone metastases, peritonitis, or pleuritis carcinomatoses. 18F-FDG uptake by gastric cancers is relatively high but does not parallel histopathologic features of malignancy. PMID- 12732671 TI - An intrapatient comparison of 99mTc-EDDA/HYNIC-TOC with 111In-DTPA-octreotide for diagnosis of somatostatin receptor-expressing tumors. AB - The aim of this study was to compare the imaging abilities of the recently developed somatostatin analog, (99m)Tc-hydrazinonicotinyl-Tyr(3)-octreotide ((99m)Tc-HYNIC-TOC [(99m)Tc-TOC]), with (111)In-diethylenediaminepentaacetic acid D-Phe(1)-octreotide ((111)In-OCT [Octreoscan]) in patients undergoing routine somatostatin receptor (SSTR) scintigraphy. METHODS: Forty-one patients (20 men, 21 women; age range, 29-75 y; mean age, 56.7 y) with either histologically proven or biologically and clinically suspected endocrine tumors were enrolled in the study. Four groups were distinguished: (a) patients being evaluated for the detection and localization of neuroendocrine tumors (n = 6), (b) tumor staging (n = 19), (c) patients being investigated to determine the SSTR status of tumor lesions (n = 11), and (d) patient follow-up studies (n = 5). Each patient received a mean activity of 150 MBq (111)In-OCT and 350-400 MBq (99m)Tc-TOC. Scintigraphy with (99m)Tc-TOC was performed 4 h after injection and scintigraphy with (111)In-OCT was performed 4 and 24 h after injection. SPECT studies of areas of interest were performed 4 h after injection for both tracers as well as at 24 h after injection for (111)In-OCT. The time interval between the studies using each tracer ranged from 2 to 22 d (mean interval, 9.3 d). RESULTS: (111)In-OCT and (99m)Tc-TOC showed an equivalent scan result in 32 patients (78%), 9 cases showed discrepancies (22%), false-negative results with (111)In-OCT were seen in 6 cases (14.6%), whereas (99m)Tc-TOC was false-positive in 2 cases (4.9%). (111)In-OCT was true-negative in both cases. The false-positive findings of the (99m)Tc-TOC studies were caused by nonspecific uptake in the bowel. In 1 case, (99m)Tc-TOC correctly identified a metastasis in the lumbar spine but both scan results were false-positive because of an inflammatory process. In 21 patients with SSTR-expressing tumors, the semiquantitative region-of-interest analysis showed that (99m)Tc-TOC achieved higher tumor-to-normal tissue ratios than (111)In-OCT. CONCLUSION: This study revealed a higher sensitivity of (99m)Tc-TOC as compared with (111)In-OCT as an imaging agent for the localization of SSTR expressing tumors. To avoid false-positive findings with (99m)Tc-OCT due to nonspecific tracer accumulation, additional scanning at 1-2 h after injection should be done. PMID- 12732670 TI - Evaluation of 111In-DTPA-folate as a receptor-targeted diagnostic agent for ovarian cancer: initial clinical results. AB - The cell-membrane folate receptor is a potential molecular target for tumor selective drug delivery, including radiolabeled folate-chelate conjugates for diagnostic imaging. We report here the initial clinical study of such an agent, (111)In-diethylenetriaminepentaacetic acid (DTPA)-folate, evaluated for diagnosis of ovarian malignancy. METHODS: Thirty-five women were enrolled in a phase I/II clinical study, with 33 completing the surgical follow-up required by the study protocol for definition of disease status. Patients either had a pathologically proven malignancy or were scheduled for surgery for suspected new ovarian cancer (n = 26), recurrent ovarian cancer (n = 5), or endometrial cancer (n = 2). (111)In-DTPA-folate was administered as an intravenous bolus, and whole-body images were obtained at 30 min, 4 h, and (for the first 19 patients) 24 h after injection; SPECT also was done at the delayed imaging times. For 19 of the patients, unlabeled free folic acid was injected before administration of (111)In DTPA-folate to also assess the impact of folate loading on tracer biodistribution. Masked and unmasked readings of the images by 2 nuclear medicine physicians were compared with the pathologic findings after surgery. RESULTS: Among 33 patients who had surgical intervention, 14 had new or recurrent malignant tumors. All of 7 newly diagnosed ovarian carcinomas were identified by both masked readers (sensitivity, 100%). The sensitivity for detection of 7 recurrent malignancies was 38% for masked readings and 85% for unmasked readings, indicating that correlation with anatomic imaging studies (CT) was highly important in diagnosis of these lesions. Eighteen of the studied patients were found to have benign masses; for this limited population, the specificity of (111)In-DTPA-folate scintigraphy was 76% and 82% for the masked and unmasked analyses, respectively. CONCLUSION: (111)In-DTPA-folate is safe, and possibly effective, for scintigraphy differentiating between malignant and benign ovarian masses. PMID- 12732672 TI - 18F-FDG PET for the diagnosis and grading of soft-tissue sarcoma: a meta analysis. AB - PET using (18)F-FDG is increasingly used for the diagnosis and grading of tumors. Several studies have been performed that evaluate the diagnostic and grading performance of (18)F-FDG PET for soft-tissue sarcoma, but each study has had a limited sample size. Therefore, we undertook a comprehensive meta-analysis of the evidence. METHODS: Relevant studies were identified from MEDLINE and EMBASE. Diagnostic and grading performance were evaluated for qualitative visualization; standard uptake value (SUV, cutoffs of 2.0 and 3.0); and metabolic rate of glucose (MRG, cutoff of 6.0 micro mol/100 g/min). Quantitative data synthesis included independent weighting of sensitivity and specificity, construction of summary receiver operating characteristic curves, and pooled analyses. RESULTS: The meta-analysis included 15 studies with 441 soft-tissue lesions (227 malignant, 214 benign). For diagnosis of malignant versus benign lesions, typical pairs of sensitivity and specificity estimates from the summary receiver operating characteristic curves were 92% and 73% for qualitative visualization; 87% and 79% for SUV 2.0; 70% and 87% for SUV 3.0; and 74% and 73% for MRG 6.0. Diagnostic performance was similar for primary and recurrent lesions. By qualitative interpretation, (18)F-FDG was positive in all intermediate/high-grade tumors (95% confidence interval [CI], 97.3%-100%), 74.4% (95% CI, 58.6%-85.9%) of low-grade tumors, and 39.3% (95% CI, 29.1%-50.3%) of benign lesions (including 11 of 12 inflammatory lesions). Using an SUV cutoff of 2.0, respective rates were 89.4% (95% CI, 79.4%-95.6%), 33.1% (95% CI, 15.6%-55.3%), and 19.1% (95% CI, 10.6%-30.5%). Limited data on comparisons with MRI and CT showed no differences against (18)F-FDG PET in diagnosing recurrent and metastatic disease. CONCLUSION: (18)F-FDG PET has very good discriminating ability in the evaluation of both primary and recurrent soft-tissue lesions. (18)F-FDG PET may be helpful in tumor grading but offers inadequate discrimination between low-grade tumors and benign lesions. PMID- 12732673 TI - Value of supranormal function and renogram patterns on 99mTc mercaptoacetyltriglycine scintigraphy in relation to the extent of hydronephrosis for predicting ureteropelvic junction obstruction in the newborn. AB - We investigated the value of supranormal function and renogram patterns on (99m)Tc-mercaptoacetyltriglycine ((99m)Tc-MAG3) scintigraphy in relation to the extent of hydronephrosis for predicting ureteropelvic junction (UPJ) obstruction in the newborn. METHODS: We studied 48 patients with prenatally diagnosed unilateral hydronephrosis that was confirmed on postnatal ultrasonography. The anteroposterior pelvic diameter (APPD) and the Society for Fetal Urology (SFU) grade were measured on 1-mo ultrasonography to assess the extent of hydronephrosis. Neonates with an APPD of or=55%) and renogram grades (1, not obstructive; 2, indeterminate; 3, obstructive; and 4, prolonged parenchymal transit) were determined on 1-mo (99m)Tc-MAG3 renography. Multivariate logistic regression analysis was performed to predict the presence of obstruction. RESULTS: Of the 48 kidneys with hydronephrosis, 19 were diagnosed as having UPJ obstruction. Twenty-nine kidneys were classified as having nonobstructive hydronephrosis. The APPD of the 19 kidneys with obstruction (24.3 +/- 9.2 mm) was significantly larger than that of the 29 kidneys without obstruction (17.5 +/- 11.2 mm; P < 0.05). The SFU grades of UPJ obstruction (2 with grade 3, 17 with grade 4) were also significantly higher than those without UPJ obstruction (grades 1-4 in 1, 6, 10, and 12 kidneys, respectively; P < 0.01). Supranormal function was present in 4 kidneys with UPJ obstruction but in none without obstruction (P < 0.001). (99m)Tc-MAG3 renography of the 19 obstructions showed grades 2, 3, and 4 renograms in 4, 8, and 7 patients, respectively. The 29 without obstruction revealed significantly lower grades (grades 1-4 in 10, 13, 5, and 1 patient, respectively; P < 0.001). Multivariate logistic regression analysis revealed that the addition of supranormal function or renogram grade in the model significantly increased the likelihood ratio in comparison with that of the SFU grade only (chi(2) = 7.73 and 9.99, respectively; P < 0.01). Of the 29 neonates with SFU grade 4, supranormal function or renogram >or= grade 4 had a positive predictive value of 90% (9/10). CONCLUSION: (99m)Tc-MAG3 renography at 1 mo after birth has a significant additive value in relation to the degree of hydronephrosis in predicting UPJ obstruction in patients with prenatally diagnosed unilateral hydronephrosis with normal function. PMID- 12732674 TI - Cause and magnitude of the error induced by oral CT contrast agent in CT-based attenuation correction of PET emission studies. AB - CT images represent essentially noiseless maps of photon attenuation at a range of 40-140 keV. Current dual-modality PET/CT scanners transform them into attenuation coefficients at 511 keV and use these for PET attenuation correction. The proportional scaling algorithms hereby used account for the different properties of soft tissue and bone but are not prepared to handle material with other attenuation characteristics, such as oral CT contrast agents. As a consequence, CT-based attenuation correction in the presence of an oral contrast agent results in erroneous PET standardized uptake values (SUVs). The present study assessed these errors with phantom measurements and patient data. METHODS: Two oral CT contrast agents were imaged at 3 different concentrations in dual modality CT and PET transmission studies to investigate their attenuation properties. The SUV error due to the presence of contrast agent in CT-based attenuation correction was estimated in 10 patients with gastrointestinal tumors as follows. The PET data were attenuation corrected on the basis of the original contrast-enhanced CT images, resulting in PET images with distorted SUVs. A second reconstruction used modified CT images wherein the CT numbers representing contrast agent had been replaced by CT values producing approximately the right PET attenuation coefficients. These CT values had been derived from the data of 10 patients imaged without a CT contrast agent. The SUV error, defined as the difference between both sets of SUV images, was evaluated in regions with oral CT contrast agent, in tumor, and in reference tissue. RESULTS: The oral CT contrast agents studied increased the attenuation for 511-keV photons minimally, even at the highest concentrations found in the patients. For a CT value of 500 Hounsfield units, the proportional scaling algorithm therefore overestimated the PET attenuation coefficient by 26.2%. The resulting SUV error in the patient studies was highest in regions containing CT contrast agent (4.4% +/- 2.8%; maximum, 11.3%), whereas 1.2% +/- 1.1% (maximum, 4.1%) was found in tumors, and 0.6% +/- 0.7% was found in the reference. CONCLUSION: The use of oral contrast agents in CT has only a small effect on the SUV, and this small effect does not appear to be medically significant. PMID- 12732675 TI - Comparison of observer variability and accuracy of different criteria for lung scan interpretation. AB - Different criteria have been advocated for the interpretation of ventilation/perfusion (V/Q) lung scans in patients with suspected pulmonary embolism (PE). Besides these predefined criteria, many physicians use an integration of the different sets of criteria and their own experience-the so called Gestalt interpretation. The purpose of this study was to evaluate interobserver variability and accuracy of 3 sets of criteria: the Hull and PIOPED (Prospective Investigation of Pulmonary Embolism Diagnosis) criteria and the Gestalt interpretation. METHODS: Two experienced observers interpreted V/Q scans of all 328 patients according to the 3 different schemes. The diagnostic classification obtained for the different sets of criteria was analyzed against the presence or absence of PE. RESULTS: The interobserver variabilities as assessed by the kappa statistics of the PIOPED and Hull criteria and for the Gestalt interpretation were 0.70 (95% confidence interval [CI], 0.64-0.76), 0.79 (95% CI, 0.73-0.85), and 0.65 (95% CI, 0.58-0.72), respectively. The differences in kappa values between the Hull and PIOPED criteria and between the Hull criteria and Gestalt interpretation were statistically significant (P < 0.05 and P < 0.001, respectively). For 16 patients (14 without PE) with a normal lung scan result according to the Hull criteria, the result according to the PIOPED criteria was low probability. For 21 patients (12 with PE), the scans were intermediate probability according to the PIOPED criteria, whereas the result with the Hull criteria was high probability. Analysis of receiver-operating characteristic curves yielded a comparable area under the curve for all sets of criteria (0.87-0.90). CONCLUSION: The Hull, PIOPED, and Gestalt interpretation of V/Q lung scans all have a good accuracy and interobserver variability. However, the reproducibility of the Hull criteria is superior in comparison with that of the other sets of criteria. PMID- 12732676 TI - The effect of nitroglycerin on myocardial blood flow in various segments characterized by rest-redistribution thallium SPECT. AB - The use of nitrates is reported to be effective in viability detection in scintigraphic perfusion imaging. The purpose of the study was to evaluate the effect of nitroglycerin (NTG) on myocardial blood flow (MBF) and coronary vascular resistance (CVR) in various segments characterized by rest redistribution (201)Tl SPECT. METHODS: Twenty-three patients with coronary artery disease underwent rest-redistribution (201)Tl SPECT and (15)O-labeled water PET at rest and after NTG spray (0.3 mg). In addition, 11 healthy volunteers were also studied using PET. RESULTS: NTG did not change global MBF in the volunteers or in the patients. In segments with normal (201)Tl uptake and in those with a severe irreversible (201)Tl defect, NTG significantly reduced MBF without changing CVR. NTG reduced CVR in segments with a reversible (201)Tl defect (141 +/- 50 to 114 +/- 29 mm Hg/[mL/min/g], P = 0.004) and in those with a mild-to moderate irreversible (201)Tl defect (165 +/- 64 to 149 +/- 60 mm Hg/[mL/min/g], P = 0.003), while maintaining MBF. CONCLUSION: NTG preferentially reduces CVR in the viable myocardium with ischemia. After NTG, tracer uptake in the ischemic myocardium will be relatively increased compared with that in the nonviable and nonischemic myocardium, leading to improvements in viability detection. PMID- 12732677 TI - Nitrates and viability: a durable affair. PMID- 12732678 TI - PET instrumentation and reconstruction algorithms in whole-body applications. AB - The aim of this work is the presentation and comparison of state-of-the-art dedicated PET systems actually available on the market, in terms of physical performance and technical features. Particular attention has been given to evaluate the whole-body performance by sensitivity, spatial resolution, dead time, noise equivalent counting rate (NECR), and scatter fraction. PET/CT systems were also included as new proposals to improve diagnostic accuracy of PET, allowing effective anatomic integration to functional data. An overview of actually implemented reconstruction algorithms is also reported to fully understand all of the factors that contribute to image quality. PMID- 12732679 TI - Acetylated low-density lipoprotein-encapsulated cholesteryl 1,3-diiopanoate glyceryl ether for the detection of atherosclerosis in rabbits. AB - A cholesteryl ester analog, cholesteryl 1,3-diiopanoate glyceryl ether (C2I), was synthesized and investigated for its potential use for the detection of atherosclerotic lesions in rabbits. METHODS: (125)I-labeled C2I was incorporated into acetylated low-density lipoprotein (AcLDL). The resultant complex, (125)I C2I-AcLDL, was injected intravenously into 2 groups of rabbits, fed cholesterol and normal chow, at a dose of 555 kBq/kg. Tissue samples were taken 24 h after injection for the biodistribution study. Atherosclerotic lesions and C2I deposition in aortic samples were examined by Sudan IV staining and autoradiography, respectively. RESULTS: The levels of C2I in blood and aortic samples in cholesterol-fed animals were 2- to 3-fold higher than those in the control group (P < 0.05). The autoradiography results correlated well with the Sudan IV staining results, indicating sites of C2I deposition superimposed on lesion sites. CONCLUSION: C2I was preferentially taken up and retained at atherosclerotic lesion sites, suggesting its potential use for the detection of early atherosclerosis. PMID- 12732680 TI - Adipocyte spatial distributions in bone marrow: implications for skeletal dosimetry models. AB - Few studies have been conducted to quantify the spatial distributions of adipocytes in the marrow cavities of trabecular bone. Nevertheless, such data are needed for the development of 3-dimensional (3D) voxel skeletal models where marrow cellularity is explicitly considered as a model parameter for dose assessment. In this investigation, bone marrow biopsies of the anterior iliac crest were examined to determine the size distribution of adipocyte cell clusters, the percentage of perimeter coverage of trabecular surfaces, and the presence or absence of adipocyte density gradients in the marrow space, all as a function of the biopsy marrow cellularity (5%-95%). METHODS: Biopsy slides from 42 patients were selected as designated by the hematopathologist as either normocellular or with no evidence of disease. Still-frame video image captures were made of 1-3 regions of interest per biopsy specimen, with subsequent image analysis of adipocyte spatial characteristics performed via a user-written MATLAB routine. RESULTS: A predictable shift was found in cluster size with decreasing marrow cellularity from single adipocytes to clusters of >or=3 cells; the percentage of 2-cell clusters remained relatively constant with changing cellularity. Also, a nonlinear increase in trabeculae perimeter coverage was found with increasing fat tissue fraction at marrow cellularities between 50% and 80%. Finally, it was demonstrated that only in the range of 20%-50% marrow cellularity was a slight gradient in adipocyte concentration indicated with adipocytes localized preferentially toward the trabecular surfaces. CONCLUSION: Electron transport simulations were conducted in 4 different 3D voxel models of trabecular bone for sources localized in the active marrow (TAM), bone volume (TBV), bone endosteum (TBE), and bone surfaces (TBS). Voxel model simulations demonstrated that absorbed fractions to active marrow given by the ICRP 30 model (MIRDOSE2) are exceedingly conservative for both TBV and TBS sources, except in the case of high-energy particles (>500 keV) at high values of marrow cellularity (>70%). Values of both phi(TAM<--TBV) and phi(TAM<--TBS) given by the Eckerman and Stabin model (MIRDOSE3) were shown to be reasonably consistent with 3D voxel model simulations at the reference cellularity of 25%, except in the case of low energy emitters (<100 keV) on the bone surfaces. PMID- 12732681 TI - Murine S factors for liver, spleen, and kidney. AB - Preclinical evaluation of new radiopharmaceuticals is performed in animal systems before testing is started in humans. These studies, often performed in murine or other rodent models, are important in understanding the relationship between absorbed dose and response, which can be translated to preclinical results for humans. In performing such calculations, either electrons are assumed to deposit all of their energy locally or idealized models of mouse anatomy are used to determine absorbed fractions. Photon contributions are generally considered negligible. To improve the accuracy of such absorbed dose calculations, mouse specific S factors for (131)I, (153)Sm, (32)P, (188)Re, and (90)Y have been generated, and the photon and electron portions have been tabulated separately. Absorbed fractions for 5 monoenergetic electrons, ranging in energy from 0.5 to 2 MeV, are also provided. METHODS: Female athymic mouse MR images were obtained on a 4.7-T MRI device. Fifteen T1-weighted, 1.5-mm-thick slices (0.5-mm gap) were collected. Using a previously developed software package, 3-dimensional Internal Dosimetry (3D-ID), organ contours were drawn to obtain a 3-dimensional representation of liver, kidneys, and spleen. Using a point-kernel convolution, the mean absorbed dose to each organ from the individual contributions of each source organ were calculated. S factor equivalent values were obtained by assuming a uniform distribution of radioactivity in each organ. Results were validated by comparing 3D-ID generated electron S factors for different-sized spheres with published data. Depending on matrix size, sphere size, and radionuclide, 1% (256(2) matrix) to 18% (64(2) matrix) agreement was obtained. RESULTS: S factor values were calculated for liver, spleen, and right and left kidneys. Cross-organ electron-absorbed fractions of up to 0.33 were obtained (e.g., (90)Y right kidney to liver). Comparisons between S factor values and values obtained assuming complete absorption of electron energy yielded differences of more than 190% ((90)Y spleen self-dose). CONCLUSION: The effect of cross-organ and self-absorbed dose is dependent on emission energy and organ geometry and should be considered in murine dose estimates. The approach used to generate these S factors is applicable to other animal systems and also to nonuniform activity distributions that may be obtained by small-animal SPECT or PET imaging or by quantitative autoradiography. PMID- 12732682 TI - Microdosimetric analysis of alpha-particle-emitting targeted radiotherapeutics using histological images. AB - The purpose of this study was to evaluate the therapeutic efficacy and limitations of alpha-particle-emitting radiolabeled compounds by means of 2 dimensional histological images and distribution of activity on a microscopic level. METHODS: A microdosimetric approach based on histological images is used to analyze the therapeutic effectiveness of alpha-particle-emitting (211)At and (213)Bi conjugated to 201B monoclonal antibody (mAb), which is reactive with murine lung blood vessels for the treatment of EMT-6 lung tumor colonies in nude mice. Autoradiography images were used to define the tissue morphology and activity distribution within lung tissues. Two animal groups were studied: Group A consisted of animals bearing small tumors (<130 micro m) and group B consisted of larger tumors (<600 micro m). Probability density functions (pdf) described the variability in average absorbed dose and survival probability among normal and tumor target cells and, in turn, were used to assess the survival fraction of tumor and normal tissue. RESULTS: The average absorbed dose to tumor cells per unit cumulated activity concentration for animals in group A was 1.10 x 10(-3) and 1.37 x 10(-3) Gy g MBq(-1) s(-1) for (211)At and (213)Bi, respectively, and for animals in group B was 3.8 x 10(-4) and 5.6 x 10(-4) Gy g MBq(-1) s(-1) for (211)At and (213)Bi, respectively. The fraction of tumor cells that received a zero absorbed dose for animals in group A was 0.04% for (213)Bi and 0.2% for (211)At and for animals in group B was 25% for (213)Bi and 31% for (211)At. Both (213)Bi- and (211)At-labeled 201B mAb were effective therapies for animals with small tumors, where predicted therapeutic effectiveness was consistent with experimental findings; however, they were ineffective for animals with larger tumors. CONCLUSION: Microdosimetric methods based on knowledge of tissue morphology and activity distribution on a small-scale level can be a useful tool for evaluating a priori the therapeutic efficacy and limitations of targeted alpha-particle endoradiotherapeutic strategies. PMID- 12732683 TI - Synthesis and biologic evaluation of monocationic asymmetric 99mTc-nitride heterocomplexes showing high heart uptake and improved imaging properties. AB - The preparation, characterization, and first biologic evaluation in rats of a novel class of monocationic (99m)Tc heart imaging agents are reported. The complexes are represented by the general formula [(99m)Tc(N)(PNP)(L)](+), where L is the monoanionic form of a dithiocarbamate ligand of the type [R(1)(R(2))-N C(=S)S](-), PNP is a diphosphine ligand of the type [(R(3))(2)P-(CH(2))(2)](2) N(R(4)), and R(1)-R(4) are organic functional groups. METHODS: The new complexes were prepared by use of both liquid and freeze-dried formulations through a 2 step procedure. The first step involved the formation of the [TcN](2+) group through the reaction of (99m)TcO(4)(-) with succinic dihydrazide as a donor of nitride nitrogen atoms (N(3-)) in the presence of Sn(2+) ions. This step was followed by the simultaneous addition to the reaction solution of the ligand PNP and of the sodium salt of the dithiocarbamate ligand (NaL) to afford the final products, [(99m)Tc(N)(PNP)(L)](+). The chemical identities of the resulting (99m)Tc complexes were determined by comparing their chromatographic properties with those of the corresponding (99g)Tc analogs prepared by use of the long-lived isotope (99g)Tc and characterized by spectroscopic and crystallographic techniques. Ex vivo biodistribution studies were conducted in rats. In vivo tomographic images of the rat heart were obtained by use of a small-animal SPECT scanner. RESULTS: The [(99m)Tc(N)(PNP)(L)](+) complexes are monocationic and possess a distorted square-pyramidal geometry in which the TcN multiple bond occupies an apical position and the diphosphine and dithiocarbamate ligands span the residual 4 coordination positions on the basal plane through the 2 phosphorus atoms and the 2 sulfur atoms, respectively. Imaging and biodistribution studies demonstrated that these radiopharmaceuticals localize selectively in the myocardium of rats and are retained in this region for a prolonged time. The kinetics of heart uptake and clearance were found to be influenced by variations in the lateral R(1)-R(4) groups. Blood and lung washouts were extremely fast. Elimination occurred mostly through the kidneys and the liver. Surprisingly, at 1 h after injection, liver activity was almost negligible. Analysis of heart-to liver and heart-to-lung uptake ratios showed that these values increased exponentially over time and became much higher than those determined for (99m)Tc sestamibi and (99m)Tc-tetrofosmin. These findings were confirmed by analysis of high-quality SPECT images collected in rats for the new complexes and compared with images obtained with (99m)Tc-sestamibi and (99m)Tc-tetrofosmin. CONCLUSION: The high myocardial uptake and the very high heart-to-lung and heart-to-liver uptake ratios indicate that the [(99m)Tc(N)(PNP)(L)](+) complexes exhibit very favorable distribution properties and could be used to obtain SPECT cardiac images with improved quality. PMID- 12732684 TI - Reproducibility of 11C-raclopride binding in the rat brain measured with the microPET R4: effects of scatter correction and tracer specific activity. AB - A new generation of commercial animal PET cameras may accelerate drug development by streamlining preclinical testing in laboratory animals. However, little information on the feasibility of using these machines for quantitative PET in small animals is available. Here we investigate the reproducibility of microPET imaging of (11)C-raclopride in the rat brain and the effects of tracer-specific activity and photon scatter correction on measures of D2 receptor (D2R) availability. METHODS: Sprague-Dawley rats (422 +/- 29 g; n = 7) were anesthetized with ketamine/xylazine and catheterized for tail vein injection of (11)C-raclopride. Each animal was positioned prone in the microPET, centering the head in the field of view. MicroPET data was collected for 60 min-starting at (11)C-raclopride injection-and binned into 24 time frames (6 x 10 s, 3 x 20 s, 8 x 60 s, 4 x 200 s, 3 x 600 s). In 3 studies, (11)C-raclopride was administered a second time in the same animal, with 2-4 h between injections. In a fourth animal, raclopride (1 mg/kg) was coinjected with (11)C-raclopride for the second injection. Three rats received a single dose of (11)C-raclopride. The range of doses for all studies was 6.11-18.54 MBq (165-501 micro Ci). The specific activity at injection was 4.07-48.1 GBq/ micro mol (0.11-1.3 Ci/ micro mol). Region-of-interest analysis was performed and the distribution volume ratio (DVR) was computed for striatum/cerebellum using sinograms uncorrected and corrected for scatter using a tail-fit method. RESULTS: Test-retest results showed that the (11)C-raclopride microPET DVR was reproducible (change in DVR = -8.3% +/- 4.4%). The average DVR from 6 rats injected with high specific activity (<4 nmol/kg) was 2.43 +/- 0.19 (coefficient of variation = 8%). The DVR for the blocking study was 1.23. The DVR depended on the mass of tracer (11)C-raclopride injected for doses >1.5 nmol/kg. Scatter fractions within the rat head were approximately 25%-45% resulting in an average increase of DVR of 3.5% (range, 0%-10%) after correction. CONCLUSION: This study shows that the (11)C-raclopride microPET-derived DVR is reproducible and suitable for studying D2R availability in the rat brain. MicroPET sensitivity was sufficient to determine reproducible DVRs from (11)C raclopride injections of 9.25 MBq (approximately 250 micro Ci). However, the effect of tracer mass on the DVR should be considered for studies using more than approximately 1-2 nmol/kg raclopride, and scatter correction has a measurable impact on the results. PMID- 12732685 TI - Novel series of 111In-labeled bombesin analogs as potential radiopharmaceuticals for specific targeting of gastrin-releasing peptide receptors expressed on human prostate cancer cells. AB - Gastrin-releasing peptide (GRP) receptors have been shown to be expressed with high densities on several types of cancer cells including prostate, breast, small cell lung, and pancreas cancers. Bombesin (BBN) has been known to bind to GRP receptors with high affinity and specificity. The aim of these studies was to develop new (111)In-labeled BBN analogs having high tumor uptake and optimal pharmacokinetics for specific targeting of human prostate cancers. METHODS: A novel series of dodecanetetraacetic acid (DOTA)-X-BBN[7-14]NH(2) (X = 0, beta Ala, 5-Ava, 8-Aoc, or 11-Aun) conjugates and their In(III)/(111)In complexes exhibiting high GRP-receptor-binding affinities were synthesized and characterized. RESULTS: In vitro competitive binding assays, using PC-3 androgen independent human prostate cancer cells, demonstrated values of <2.5 nmol/L for inhibitory concentration of 50% for analogs with beta-Ala, 5-Ava, and 8-Aoc spacers. In vivo biodistribution studies of the (111)In-DOTA-X-BBN[7-14]NH(2) conjugates performed on CF-1 mice at 1 h after injection have revealed that the uptake of radioactivity in the pancreas, a GRP-receptor-expressing tissue, increased as a function of hydrocarbon spacer length (i.e., from 0.20 +/- 0.04 percentage injected dose [%ID] per gram for the analog with no spacer to a maximum of 26.97 +/- 3.97 %ID/g for the analog with 8-Aoc spacer). The radioactivity was cleared efficiently from the blood pool by excretion mainly through the renal/urinary pathway (e.g., 71.6 +/- 1.8 %ID at 1 h after injection for 8-Aoc spacer analog). In vivo pharmacokinetic studies of the (111)In-DOTA-8 Aoc-BBN[7-14]NH(2) conjugate conducted on PC-3 human prostate cancer-derived xenografts in SCID mice showed a specific uptake of radioactivity in tumor, with 3.63 +/- 1.11 %ID/g observed at 1 h after injection. High tumor-to-blood and tumor-to-muscle ratios of approximately 6:1 and 45:1, respectively, were achieved at 1 h after injection. Relative to the radioactivity observed in the tumor at 1 h after injection, 43%, 19%, and 9% of the radioactivity was retained at, respectively, 24, 48, and 72 h after injection. CONCLUSION: These studies showed that radiometallated DOTA-X-BBN[7-14]NH(2) constructs with hydrocarbon spacers ranging from 5 to 8 carbon atoms are feasible candidates for further development as diagnostic and therapeutic radiopharmaceuticals for patients with GRP-positive cancers. PMID- 12732686 TI - Potential interference of agents on radioiodide thyroid uptake in the euthyroid rat. AB - The objective of this research was to investigate the merits of controlled studies with euthyroid rats as a means of determining the influence of dose and time after administration of agents that may interfere with radioiodide uptake in the thyroid. METHODS: Potassium iodide (KI), propylthiouracil (PTU), diatrizoate meglumine, and iohexol were selected to represent interfering agents. Two dose levels per agent were investigated. Doses used were 1 and 2 mg/kg of body weight for KI, 3.5 and 7 mg/kg of body weight for PTU, 1 mL/kg (282 mg I/kg) and 2 mL/kg (564 mg I/kg) of body weight for diatrizoate meglumine, and 1 mL/kg (300 mg I/kg) and 2 mL/kg (600 mg I/kg) of body weight for iohexol. The 24-h radioiodide thyroid uptake was determined after (131)I was given at 1, 8, 15, and 22 d after administration of interfering agents. RESULTS: The percentage radioiodide uptake value for the thyroid decreased significantly compared with controls for all agents and both doses on day 1 but returned to control levels by day 22 for all agents and both doses The time to return to normal varied between agents and doses. CONCLUSION: We conclude that the interfering agent, the dose given, and the length of time after administration influence the potential for an agent to affect radioiodide uptake in the thyroid. Further studies with the rat, preferably hyperthyroid, would be beneficial in generating data to reduce confusing contradictory information on the length and severity of interference of agents in radioiodide thyroid studies. PMID- 12732687 TI - Investigating dopaminergic neurotransmission with 123I-FP-CIT SPECT: comparability of modern SPECT systems. AB - With (123)I-labeled N-omega-fluoropropyl-2-beta-carbomethoxy-3-beta-(4 iodophenyl)nortropane (FP-CIT) SPECT increasingly gaining access into routine patient care, the comparability of the results of different SPECT systems in the quantification of receptor binding is important for accurate clinical decision making and the translation of imaging results between institutions (e.g., as part of multicenter therapy trials). METHODS: In a series of studies using phantoms (containing target cylinders of 2- and 2.8-cm diameter) and (123)I-FP-CIT patient studies (n = 49), we compared target-to-background (T/BG) and (123)I-FP-CIT striatal uptake ratios recovered by a conventional triple-head SPECT system and a dedicated high-resolution brain SPECT system. All patient studies were acquired on both SPECT systems successively (<15-min interscan gap) using a single injection protocol (group A [n = 20] was first scanned on the triple-head SPECT system, and group B [n = 29] was first scanned on the dedicated brain SPECT system). RESULTS: In phantom studies, the T/BG ratios recovered by both systems correlated strongly with the true T/BG ratios (R(2) > 0.96), with the linear regression slopes being 0.86-1.17 and 0.41-0.52 (less steep for smaller target size and lower T/BG ratios) for the dedicated brain SPECT and the triple-head SPECT system, respectively. Although both systems yielded markedly different results, they showed a high linear correlation with each other (R(2) > 0.95, no significant effect from target size). In (123)I-FP-CIT patient studies, a similar linear intersystem correlation was found (R(2) [A/B] = 0.79/0.80, 0.52/0.68, and 0.83/0.85 for the uptake ratios of the striatum, caudate, and putamen, respectively, to the occipital reference region). CONCLUSION: A linear transformation of striatal uptake ratios between different SPECT systems appears to be achievable for ligands such as (123)I-FP-CIT. An evaluation is needed of whether the present method can do this with sufficient accuracy for clinical purposes or whether methodologic adaptations are necessary. Proper study timing has to be ensured. PMID- 12732688 TI - Procedure guideline for brain death scintigraphy. PMID- 12732689 TI - Whole-body thyroid tumor 123I scintigraphy. PMID- 12732690 TI - Administration guidelines for radioimmunotherapy of non-Hodgkin's lymphoma with 90Y-labeled anti-CD20 monoclonal antibody. PMID- 12732691 TI - Sample size estimation: how many individuals should be studied? AB - The number of individuals to include in a research study, the sample size of the study, is an important consideration in the design of many clinical studies. This article reviews the basic factors that determine an appropriate sample size and provides methods for its calculation in some simple, yet common, cases. Sample size is closely tied to statistical power, which is the ability of a study to enable detection of a statistically significant difference when there truly is one. A trade-off exists between a feasible sample size and adequate statistical power. Strategies for reducing the necessary sample size while maintaining a reasonable power will also be discussed. PMID- 12732692 TI - Angioplasty, C-reactive protein, and the patient at risk. PMID- 12732693 TI - Prediction of restenosis after carotid artery stent implantation. PMID- 12732694 TI - Hemorrhagic shearing lesions in children and adolescents with posttraumatic diffuse axonal injury: improved detection and initial results. AB - PURPOSE: To compare the effectiveness of a high-spatial-resolution susceptibility weighted (SW) magnetic resonance (MR) imaging technique with that of a conventional gradient-recalled-echo (GRE) MR imaging technique for detection of hemorrhage in children and adolescents with diffuse axonal injury (DAI). MATERIALS AND METHODS: Seven young patients with a mean Glasgow Coma Scale score of 7 +/- 4 (SD) at admission were imaged a mean of 5 days +/- 3 after injury. High-spatial-resolution three-dimensional GRE imaging performed with postprocessing by using a normalized phase mask was compared with conventional GRE MR imaging. The total and mean values of lesion number and apparent hemorrhage volume load determined with both examinations were compared. Mean values were compared by using paired t test analysis. Differences were considered to be significant at P < or =.05. RESULTS: Hemorrhagic lesions were much more visible on SW MR images than on conventional GRE MR images. SW MR imaging depicted 1,038 hemorrhagic DAI lesions with an apparent total hemorrhage volume of 57,946 mm3. GRE MR imaging depicted 162 lesions with an apparent total hemorrhage volume of 28,893 mm3. SW MR imaging depicted a significantly higher mean number of lesions in all patients than did GRE MR imaging, according to results of visual (P =.004) and computer (P =.004) counting analyses. The mean hemorrhage volume load for all patients also was significantly greater (P =.014) by using SW MR imaging according to computer analysis. SW MR imaging appeared to depict much smaller hemorrhagic lesions than GRE MR imaging. The majority (59%) of individual hemorrhagic DAI lesions seen on SW MR images were small in area (<10 mm(2)), whereas the majority (43%) of lesions seen on GRE images were larger in area (10-20 mm(2)). CONCLUSION: SW MR imaging depicts significantly more small hemorrhagic lesions than does conventional GRE MR imaging and therefore has the potential to improve diagnosis of DAI. PMID- 12732695 TI - Morphologic predictors of lymph node status in rectal cancer with use of high spatial-resolution MR imaging with histopathologic comparison. AB - PURPOSE: To evaluate signal intensity and border characteristics of lymph nodes at high-spatial-resolution magnetic resonance (MR) imaging in patients with rectal cancer and to compare these findings with size in prediction of nodal status. MATERIALS AND METHODS: Forty-two patients who underwent total mesorectal excision of the rectum to determine if they had rectal carcinoma were studied with preoperative thin-section MR imaging. Lymph nodes were harvested from 42 transversely sectioned surgical specimens. The slice of each lymph node was carefully matched with its location on the corresponding MR images. Nodal size, border contour, and signal intensity on MR images were characterized and related to histologic involvement with metastases. Differences in sensitivity and specificity with border or signal intensity were calculated with CIs by using method 10 of Newcombe. RESULTS: Of the 437 nodes harvested, 102 were too small (<3 mm) to be depicted on MR images, and only two of these contained metastases. In 15 (68%) of 22 patients with nodal metastases, the size of normal or reactive nodes was equal to or greater than that of positive nodes in the same specimen. Fifty-one nodes were above the area imaged, and seven of these contained metastases. The diameter of benign and malignant nodes was similar; therefore, size was a poor predictor of nodal status. If a node was defined as suspicious because of an irregular border or mixed signal intensity, a superior accuracy was obtained and resulted in a sensitivity of 51 (85%) of 60 (95% CI: 74%, 92%) and a specificity of 216 (97%) of 221 (95% CI: 95%, 99%). CONCLUSION: Prediction of nodal involvement in rectal cancer with MR imaging is improved by using the border contour and signal intensity characteristics of lymph nodes instead of size criteria. PMID- 12732696 TI - Colorectal cancer screening with CT colonography, colonoscopy, and double contrast barium enema examination: prospective assessment of patient perceptions and preferences. AB - PURPOSE: To prospectively assess and compare perceptions of and preferences for computed tomographic (CT) colonography, colonoscopy, and double-contrast barium enema examination (DCBE) by asymptomatic patients undergoing colorectal cancer screening. MATERIALS AND METHODS: A total of 696 asymptomatic patients at higher than-average risk undergoing colorectal cancer screening were consecutively recruited to undergo both CT colonography and colonoscopy (group 1), and a like group of 617 patients was separately recruited to undergo both CT colonography and DCBE (group 2). Standard bowel preparations were different between the groups undergoing colonoscopy and DCBE. Each patient completed a questionnaire that assessed preparation inconvenience and discomfort, examination discomfort, willingness to repeat examinations, and examination preference. Survey results were compared for significance by using the Wilcoxon rank sum or chi2 test. RESULTS: The majority of patients considered the preparation to be uncomfortable (group 1, 460 of 515 [89%]; group 2, 482 of 538 [90%]) and inconvenient (group 1, 393 of 502 [78%]; group 2, 427 of 527 [81%]). Reported discomfort was similar at CT colonography and colonoscopy (P =.63) but was less at CT colonography than at DCBE (P <.001). Patients experienced significantly less discomfort than expected at both CT colonography and colonoscopy but not at DCBE. Patients' willingness to undergo frequent rescreening was significantly greater for CT colonography than for either colonoscopy or DCBE. The acceptable time interval between screenings was significantly shorter for all examinations if the bowel preparation could be avoided. Overall, patients preferred CT colonography to colonoscopy (group 1, 72.3% vs 5.1%; P <.001) or to DCBE (group 2, 97.0% vs 0.4%; P <.001). CONCLUSION: Patients undergoing colorectal cancer screening prefer CT colonography to both colonoscopy and DCBE. The majority of patients experience discomfort and inconvenience with cathartic bowel preparation. PMID- 12732697 TI - Assessment of inter- and intraobserver agreement between intravascular US and aortic angiography of thoracic aortic injury. AB - PURPOSE: To compare inter- and intraobserver agreement at thoracic aortic angiography with that at intravascular ultrasonography (US) in the work-up of patients suspected of having thoracic aortic injury. MATERIALS AND METHODS: Three blinded readers performed a retrospective review of 95 thoracic aortic angiograms and 23 intravascular US images obtained in patients suspected of having traumatic aortic injury. Inter- and intraobserver agreement in the interpretation of the thoracic aortic angiograms and intravascular US images were determined by using Cohen kappa statistics. In addition, differences among demographic groups were evaluated by using bivariate analysis. RESULTS: Interobserver agreement was lowest in the interpretation of indeterminate angiograms (kappa = 0.55) and highest in the interpretation of determinate angiograms (kappa = 0.71). In contrast, interobserver agreement in the interpretation of intravascular US images was excellent. For all groups, intraobserver agreement in the interpretation of aortic angiograms was substantial and overall agreement was good (kappa = 0.88). Intraobserver agreement in the interpretation of intravascular US images was excellent (kappa = 1.00). Differences among demographic groups were not found to be significant. CONCLUSION: Intravascular US is an adjunct to aortic angiography and yields excellent overall inter- and intraobserver agreement. Subgroups of patients who are suspected of having aortic injury and have indeterminate angiograms may benefit from undergoing intravascular US. PMID- 12732698 TI - The gloved finger sign. PMID- 12732699 TI - Pulmonary embolism: optimization of small pulmonary artery visualization at multi detector row CT. AB - PURPOSE: To compare the frequency of well-visualized pulmonary arteries according to anatomic level by using different collimation with single- and multi-detector row computed tomography (CT) in patients suspected of having acute pulmonary embolism. MATERIALS AND METHODS: Sixty patients were examined with one of three techniques (20 patients each). Group 1 was examined with single-detector row CT with 3-mm collimation and 1.3-1.6 pitch; groups 2 and 3, with multi-detector row CT with 2.5- and 1.25-mm collimation, respectively. Three thoracic radiologists independently reviewed examination findings to determine if each main, lobar, segmental, and subsegmental artery was well visualized for presence of pulmonary embolism. chi2 tests were performed. For well-visualized vessels, the presence and/or absence of pulmonary embolism was recorded and kappa statistic was determined. RESULTS: Reader 1 scored 95% (114 of 120), 96% (115 of 120), and 99% (119 of 120) of lobar arteries (P >.05); 76% (304 of 400), 86% (346 of 400), and 91% (363 of 400) of segmental arteries (P <.001); and 37% (300 of 800), 56% (448 of 800), and 76% (608 of 800) of subsegmental arteries as well visualized (P <.001) using techniques 1, 2, and 3, respectively. Reader 2 scored 97% (116 of 120), 95% (114 of 120), and 99% (119 of 120) of lobar arteries (P >.05); 77% (308 of 400), 87% (349 of 400), and 93% (371 of 400) of segmental arteries (P <.001); and 39% (310 of 800), 53% (422 of 800), and 78% (621 of 800) of subsegmental arteries (P <.001) as well visualized using techniques 1, 2, and 3, respectively. Reader 3 scored 86% (103 of 120), 82% (98 of 120), and 91% (109 of 120) of lobar arteries (P >.05); 63% (252 of 400), 70% (280 of 400), and 85% (339 of 400) of segmental arteries (P <.001); and 39% (310 of 800), 56% (451 of 800), and 71% (572 of 800) of subsegmental arteries (P <.001) as well visualized using techniques 1, 2, and 3, respectively. Sixteen patients had pulmonary embolism. Interobserver agreement for detection of pulmonary embolism was significantly better for segmental and subsegmental arteries for all readers with technique 3 (segmental, kappa = 0.79-0.80; subsegmental, kappa = 0.71-0.76) than that with technique 1 (segmental, kappa = 0.47-0.75; subsegmental, kappa = 0.28-0.54). CONCLUSION: Multi-detector row CT at 1.25-mm collimation significantly improves visualization of segmental and subsegmental arteries and interobserver agreement in detection of pulmonary embolism. PMID- 12732700 TI - Computer-aided diagnosis to distinguish benign from malignant solitary pulmonary nodules on radiographs: ROC analysis of radiologists' performance--initial experience. AB - PURPOSE: To evaluate radiologists' performance for determining a distinction between benign and malignant pulmonary nodules on chest radiographs without and with use of a computer-aided diagnosis scheme. MATERIALS AND METHODS: Fifty-three chest radiographs that depicted 31 primary lung cancers and 22 benign nodules were used. The likelihood measure of malignancy for each nodule was determined by using an automated computerized scheme. Sixteen radiologists (nine attending radiologists and seven radiology residents) participated in an observer study in which cases were interpreted first without and then with use of the scheme. The radiologists' performance was evaluated with receiver operating characteristic analysis. RESULTS: The mean area under the best-fit binormal receiver operating characteristic curve plotted in the unit square (Az) values of radiologists who interpreted images without and with the scheme were 0.743 and 0.817, respectively. The performance of radiologists was improved significantly when the scheme was used (P =.002). However, the performance (Az = 0.889) of the computer alone exceeded these results by a substantial margin. The average change in radiologists' confidence level for interpretation without and with the scheme was highly correlated (r = 0.845) with the likelihood measure of malignancy, which was presented as computer output. CONCLUSION: This scheme for computer-aided diagnosis has the potential to improve the accuracy of radiologists' performance in the classification of benign and malignant solitary pulmonary nodules. PMID- 12732702 TI - Case 58: giant cell arteritis. PMID- 12732703 TI - Recurrence in autogenous myocutaneous flap reconstruction after mastectomy for primary breast cancer: US diagnosis. AB - PURPOSE: To assess the value of ultrasonography (US) and US-guided fine-needle aspiration biopsy (FNAB) in the detection and diagnosis of recurrent cancer in breasts reconstructed with autogenous myocutaneous flaps after mastectomy for primary breast cancer and to describe the US appearances of recurrence in the reconstructed breast. MATERIALS AND METHODS: Between July 1994 and March 2001, US of the reconstructed breast was performed in 20 women with autogenous myocutaneous flap reconstruction. US findings were correlated with clinical and mammographic findings. US-guided FNAB of 25 (64%) of the 39 recurrent cancers depicted at US was performed. RESULTS: Twenty-one (54%) of the 39 recurrent cancers depicted at US were clinically occult. Mammography performed in 12 of the 20 patients with reconstructed breasts depicted 14 (56%) of the 25 recurrent cancers that were detected at US in these patients. US-guided FNAB helped to establish a definitive diagnosis of recurrent breast carcinoma in 24 (96%) of the 25 tumor specimens sampled. CONCLUSION: US and US-guided FNAB are valuable for the assessment of both palpable and clinically occult recurrent breast cancers in autogenous myocutaneous flap breast reconstructions. PMID- 12732704 TI - US of the anterior bundle of the ulnar collateral ligament: findings in five cadaver elbows with MR arthrographic and anatomic comparison--initial observations. AB - PURPOSE: To characterize the ultrasonographic (US) appearance of the anterior bundle of the ulnar collateral ligament of the elbow by comparing US images with magnetic resonance (MR) arthrograms and anatomic slices. MATERIALS AND METHODS: The ulnar collateral ligament in four cadavers (eight elbows) was blindly evaluated with US by one musculoskeletal radiologist with experience in musculoskeletal US. These results were compared with standard arthrograms, MR arthrograms, and anatomic slices by consensus reading of two musculoskeletal radiologists. The criteria for an abnormal ulnar collateral ligament included contrast material extension into the ligament or fiber discontinuity, as documented by MR arthrography and anatomic slices. RESULTS: Standard arthrography, MR arthrography, and anatomic slices demonstrated the ulnar collateral ligament to be unequivocally normal in three specimens and abnormal in two. The remaining three elbows did not meet the criteria for classification as either normal or abnormal, and thus they were excluded from the study. With US, the normal ulnar collateral ligament was fibrillar and hyperechoic between the medial epicondyle and proximal ulna. In the two abnormal cases, abnormal hypoechogenicity and ligament fiber disruption were noted. In addition, the proximal aspect of the ulnar collateral ligament varied from a cordlike structure to a broad attachment to the undersurface of the medial epicondyle with variable fat. CONCLUSION: In this small sample, the anterior bundle of the ulnar collateral ligament is identified with US by its hyperechoic and compact fibrillar echotexture. The proximal attachment of the ulnar collateral ligament has a variable appearance. Hypoechogenicity and fiber disruption indicated ulnar collateral ligament abnormality. PMID- 12732705 TI - Effectiveness of US breast cancer screening remains to be demonstrated. PMID- 12732706 TI - Breast US screening: effect of prevalent cancers. PMID- 12732707 TI - Screening breast US. PMID- 12732708 TI - Generalists versus specialists in mammography. PMID- 12732709 TI - Meta-analysis of transarterial embolization in patients with unresectable hepatocellular carcinoma. PMID- 12732710 TI - More whistles and bells for fly hearing. PMID- 12732711 TI - Alcohol potently inhibits the kainate receptor-dependent excitatory drive of hippocampal interneurons. AB - Kainate receptors (KA-Rs) are members of the glutamate-gated family of ionotropic receptors, which also includes N-methyl-d-aspartate (NMDA) and alpha-amino-3 hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors. KA-Rs are important modulators of interneuron excitability in the CA1 region of the hippocampus. Activation of these receptors enhances interneuron firing, which robustly increases spontaneous inhibitory postsynaptic currents in pyramidal neurons. We report here that ethanol (EtOH) potently inhibits this KA-R-mediated effect at concentrations as low as those that can be achieved in blood after the ingestion of just 1-2 drinks (5-10 mM). Pressure application of kainate, in the presence of AMPA and NMDA receptor antagonists, evoked depolarizing responses in interneurons that triggered repetitive action potential firing. EtOH potently inhibited these responses to a degree that was sufficient to abolish action potential firing. This effect appears to be specific for KA-Rs, as EtOH did not affect action potential firing triggered by AMPA receptor-mediated depolarizing responses. Importantly, EtOH inhibited interneuron action potential firing in response to KA R activation by synaptically released glutamate, suggesting that our findings are physiologically relevant. KA-R-dependent modulation of glutamate release onto pyramidal neurons was not affected by EtOH. Thus, EtOH increases excitability of pyramidal neurons indirectly by inhibiting the KA-R-dependent drive of gamma aminobutyric acid (GABA)ergic interneurons. We postulate that this effect may explain, in part, some of the paradoxical excitatory actions of this widely abused substance. The excitatory actions of EtOH may be perceived as positive by some individuals, which could contribute to the development of alcoholism. PMID- 12732712 TI - Sympatric speciation as a consequence of male pregnancy in seahorses. AB - The phenomenon of male pregnancy in the family Syngnathidae (seahorses, pipefishes, and sea dragons) undeniably has sculpted the course of behavioral evolution in these fishes. Here we explore another potentially important but previously unrecognized consequence of male pregnancy: a predisposition for sympatric speciation. We present microsatellite data on genetic parentage that show that seahorses mate size-assortatively in nature. We then develop a quantitative genetic model based on these empirical findings to demonstrate that sympatric speciation indeed can occur under this mating regime in response to weak disruptive selection on body size. We also evaluate phylogenetic evidence bearing on sympatric speciation by asking whether tiny seahorse species are sister taxa to large sympatric relatives. Overall, our results indicate that sympatric speciation is a plausible mechanism for the diversification of seahorses, and that assortative mating (in this case as a result of male parental care) may warrant broader attention in the speciation process for some other taxonomic groups as well. PMID- 12732713 TI - Subcellular localization of yeast ribonucleotide reductase regulated by the DNA replication and damage checkpoint pathways. AB - The fidelity of DNA replication and repair processes is critical for maintenance of genomic stability. Ribonucleotide reductase (RNR) catalyzes the rate-limiting step in dNTP production and thus plays an essential role in DNA synthesis. The level and activity of RNR are highly regulated by the cell cycle and DNA damage checkpoints, which maintain optimal dNTP pools required for genetic fidelity. RNRs are composed of a large subunit that binds the nucleoside diphosphate substrates and allosteric effectors and a small subunit that houses the di-iron tyrosyl radical cofactor essential for the reduction process. In Saccharomyces cerevisiae, there are two large subunits (Rnr1 and Rnr3) and two small subunits (Rnr2 and Rnr4). Here we report the subcellular localization of Rnr1-4 during normal cell growth and the redistribution of Rnr2 and Rnr4 in response to DNA damage and replicational stress. During the normal cell cycle, Rnr1 and Rnr3 are predominantly localized to the cytoplasm and Rnr2 and Rnr4 are predominantly present in the nucleus. Under genotoxic stress, Rnr2 and Rnr4 become redistributed to the cytoplasm in a checkpoint-dependent manner. Subcellular redistribution of Rnr2 and Rnr4 can occur in the absence of the transcriptional induction of the RNR genes after DNA damage and likely represents a posttranslational event. These results suggest a mechanism by which DNA damage checkpoint modulates RNR activity through the temporal and spatial regulation of its subunits. PMID- 12732714 TI - Mind-altering miniature neurotransmitter release? PMID- 12732716 TI - Guanylate cyclase-activating protein (GCAP) 1 rescues cone recovery kinetics in GCAP1/GCAP2 knockout mice. AB - Mediated by guanylate cyclase-activating proteins (GCAPs), cytoplasmic Ca2+ levels regulate the activity of photoreceptor guanylate cyclase (GC) and the synthesis of cGMP, the internal transmitter of phototransduction. When GCAP1 is expressed in transgenic mice on a GCAP null background, it restores the wild-type flash responses in rod photoreceptors. In this communication, we explored the role of GCAP1 in cone photoreceptors by using electroretinograms (ERGs). Under cone isolation conditions, ERGs recorded from mice lacking both GCAP1 and GCAP2 had normal amplitudes of the saturated a-wave and b-wave. However, recordings from these mice demonstrated a widened b-wave and increased sensitivity of both M and UV-cone systems. Paired-flash ERGs revealed a delayed recovery of both the cone driven b-wave and a-wave and suggest that the delay originated from the photoreceptors. To test whether GCAP1 could restore normal cone response recovery, mice that expressed only transgenic GCAP1 in the absence of wild-type GCAP expression were tested. Immunohistochemical analysis demonstrated that cones of these mice expressed high levels of GCAP1. Paired-flash ERGs showed that the recovery of the cone-driven a-wave was restored to normal, whereas recovery of the cone-driven b-wave was slightly faster than that observed in wild-type mice. These studies reveal that, similar to rods, deletion of GCAP1 and GCAP2 delays the recovery of light responses in cones and GCAP1 restores the recovery of cone responses in the absence of GCAP2. PMID- 12732715 TI - Antagonistic control of oxidative stress-induced cell death in Arabidopsis by two related, plant-specific zinc finger proteins. AB - The most familiar form of plant programmed cell death is the hypersensitive response (HR) associated with successful plant immune responses. HR is preceded by an oxidative burst and the generation of both reactive oxygen intermediates (ROI) and NO. The Arabidopsis LSD1 gene encodes a negative regulator of plant programmed cell death that meets several criteria for a regulator of processes relevant to ROI management during pathogen responses. Here we demonstrate that a highly conserved LSD1 paralogue, LOL1, acts as a positive regulator of cell death. Manipulation of LOL1 expression alters both the superoxide-dependent, runaway cell death phenotype of lsd1 plants and the normal HR. We also show that LSD1 and LOL1 have antagonistic effects on copper-zinc superoxide dismutase accumulation, consistent with functions in cell death control via maintenance of ROI homeostasis. PMID- 12732717 TI - Secretory leukocyte protease inhibitor promotes the tumorigenic and metastatic potential of cancer cells. AB - Because of their ability to inhibit proteases, protease inhibitors have generally been considered to counteract tumor progression and metastasis. However, expression of serine protease inhibitors (SPIs) in tumors is often associated with poor prognosis of cancer patients. Moreover, there is growing evidence that SPIs may even promote malignancy of cancer cells, opening new avenues for their use as biomarkers in malignancy. To isolate cancer promoting genes, we applied the suppression subtractive hybridization method to low-malignant Lewis Lung Carcinoma 3LL-S versus high-malignant 3LL-S-sc cells. This resulted in the identification of the SPI secretory leukocyte protease inhibitor (SLPI), as one of the genes whose expression was higher in 3LL-S-sc than in 3LL-S cells. By stable transfection of 3LL-S cells with mouse or human SLPI, we demonstrated that elevated levels of SLPI expression increased both the tumorigenicity and lung colonizing potential of 3LL-S cells. Moreover, we showed that this function of SLPI depended on its protease inhibitory capacity. Our results also reveal that although SLPI enhanced the proliferation of 3LL-S cells in vitro, its promalignant activity in vivo was not solely due to its effect on cell proliferation. In this study, we report a causal role for SLPI in the malignant behavior of cancer cells, underscoring the potential malignancy-promoting activities of SPIs. PMID- 12732718 TI - Mesenchymal progenitor self-renewal deficiency leads to age-dependent osteoporosis in Sca-1/Ly-6A null mice. AB - The cellular and molecular mechanisms that underlie age-dependent osteoporosis, the most common disease in the Western Hemisphere, are poorly understood in part due to the lack of appropriate animal models in which to study disease progression. Here, we present a model that shows many similarities to the human disease. Sca-1, well known for its expression on hematopoietic stem cells, is present on a subset of bone marrow stromal cells, which potentially include mesenchymal stem cells. Longitudinal studies showed that Sca-1(-/-) mice undergo normal bone development but with age exhibit dramatically decreased bone mass resulting in brittle bones. In vivo and in vitro analyses demonstrated that Sca-1 is required directly for the self-renewal of mesenchymal progenitors and indirectly for the regulation of osteoclast differentiation. Thus, defective mesenchymal stem or progenitor cell self-renewal may represent a previously uncharacterized mechanism of age-dependent osteoporosis in humans. PMID- 12732719 TI - Caspase-mediated processing of the Drosophila NF-kappaB factor Relish. AB - The NF-kappaB-like transcription factor Relish plays a central role in the innate immune response of Drosophila. Unlike other NF-kappaB proteins, Relish is activated by endoproteolytic cleavage to generate a DNA-binding Rel homology domain and a stable IkappaB-like fragment. This signal-induced endoproteolysis requires the activity of several gene products, including the IkappaB kinase complex and the caspase Dredd. Here we used mutational analysis and protein microsequencing to demonstrate that a caspase target site, located in the linker region between the Rel and the IkappaB-like domain, is the site of signal dependent cleavage. We also show physical interaction between Relish and Dredd, suggesting that Dredd indeed is the Relish endoprotease. In addition to the caspase target site, the C-terminal 107 aa of Relish are required for endoproteolysis and signal-dependent phosphorylation by the Drosophila IkappaB kinase beta. Finally, an N-terminal serine-rich region in Relish and the PEST domain were found to negatively regulate Relish activation. PMID- 12732720 TI - Behavioral, neurochemical, and electrophysiological characterization of a genetic mouse model of depression. AB - Depression is a multifactorial illness and genetic factors play a role in its etiology. The understanding of its physiopathology relies on the availability of experimental models potentially mimicking the disease. Here we describe a model built up by selective breeding of mice with strikingly different responses in the tail suspension test, a stress paradigm aimed at screening potential antidepressants. Indeed, "helpless" mice are essentially immobile in the tail suspension test, as well as the Porsolt forced-swim test, and they show reduced consumption of a palatable 2% sucrose solution. In addition, helpless mice exhibit sleep-wakefulness alterations resembling those classically observed in depressed patients, notably a lighter and more fragmented sleep, with an increased pressure of rapid eye movement sleep. Compared with "nonhelpless" mice, they display higher basal seric corticosterone levels and lower serotonin metabolism index in the hippocampus. Remarkably, serotonin(1A) autoreceptor stimulation induces larger hypothermia and inhibition of serotoninergic neuronal firing in the nucleus raphe dorsalis in helpless than in nonhelpless mice. Thus, helpless mice exhibit a decrease in serotoninergic tone, which evokes that associated with endogenous depression in humans. Finally, both the behavioral impairments and the serotoninergic dysfunction can be improved by chronic treatment with the antidepressant fluoxetine. The helpless line of mice may provide an opportunity to approach genes influencing susceptibility to depression and to investigate neurophysiological and neurochemical substrates underlying antidepressant effects. PMID- 12732721 TI - Rb function in extraembryonic lineages suppresses apoptosis in the CNS of Rb deficient mice. AB - Retinoblastoma (Rb)-deficient embryos show severe defects in neurogenesis, erythropoiesis, and lens development and die at embryonic day 14.5. Our recent results demonstrated a drastic disorganization of the labyrinth layer in the placenta of Rb-deficient embryos, accompanied by reduced placental transport function. When these Rb-/- embryos were supplied with a wild-type placenta by using either tetraploid aggregation or genetic approaches, animals survived until birth. Here we analyze the role of extraembryonic Rb in regulating proliferation, apoptosis, and differentiation in the rescued animals at different developmental stages. Many of the neurological and erythroid abnormalities thought to be responsible for the embryonic lethality of Rb-/- animals, including the ectopic apoptosis in the CNS, were virtually absent in rescued Rb-/- pups. However, rescued animals died at birth with severe skeletal muscle defects. Like in Rb knockout embryos, rescued animals showed a marked increase in DNA replication and cell division in the CNS. In sharp contrast, the typical widespread neuronal apoptosis was absent in Rb-deficient embryos reconstituted with a normal placenta. In lens fiber cells, however, the inappropriate proliferation and apoptosis that is normally observed in Rb-/- embryos continued unabated in rescued animals. These results demonstrate that Rb function in extraembryonic lineages plays an important role in the survival of neuronal cells and in the differentiation of the erythroid lineage, providing mechanistic insight into the cell autonomous and nonautonomous functions of Rb during development. PMID- 12732723 TI - A quantitative model for transforming reflectance spectra into the Munsell color space using cone sensitivity functions and opponent process weights. AB - This article presents a computational model of the process through which the human visual system transforms reflectance spectra into perceptions of color. Using physical reflectance spectra data and standard human cone sensitivity functions we describe the transformations necessary for predicting the location of colors in the Munsell color space. These transformations include quantitative estimates of the opponent process weights needed to transform cone activations into Munsell color space coordinates. Using these opponent process weights, the Munsell position of specific colors can be predicted from their physical spectra with a mean correlation of 0.989. PMID- 12732722 TI - Inducible expression of keratinocyte growth factor (KGF) in mice inhibits lung epithelial cell death induced by hyperoxia. AB - Oxidant-induced injury to the lung is associated with extensive damage to the lung epithelium. Instillation of keratinocyte growth factor (KGF) in the lungs of animals protects animals from oxidant-induced injury but the mechanism of protection is not well understood. An inherent problem in studying KGF function in vivo has been that constitutive overexpression of KGF in the lung causes embryonic lethality with extensive pulmonary malformation. Here we report the development of a stringently regulated, tetracycline-inducible, lung-specific transgenic system that allows regulated expression of KGF in the lung without causing developmental abnormalities from leaky KGF expression. By using this system, we show that exposure of KGF-expressing mice to hyperoxia protects the lung epithelium but not the endothelium from cell death in accordance with the selective expression of KGF receptor on epithelial and not on endothelial cells. Investigations of KGF-induced cell survival pathways revealed KGF-induced activation of the multifunctional pro-survival Akt signaling axis both in vitro and in vivo. Inhibition of KGF-induced Akt activation by a dominant-negative mutant of Akt blocked the KGF-mediated protection of epithelial cells exposed to hyperoxia. KGF-induced Akt activation may play an important role in inhibiting lung alveolar cell death thereby preserving the lung architecture and function during oxidative stress. PMID- 12732724 TI - Ultra-high-pressure inactivation of prion infectivity in processed meat: a practical method to prevent human infection. AB - Bovine spongiform encephalopathy contamination of the human food chain most likely resulted from nervous system tissue in mechanically recovered meat used in the manufacture of processed meats. We spiked hot dogs with 263K hamster-adapted scrapie brain (10% wtwt) to produce an infectivity level of approximately 9 log(10) mean lethal doses (LD(50)) per g of paste homogenate. Aliquots were subjected to short pressure pulses of 690, 1,000, and 1,200 MPa at running temperatures of 121-137 degrees C. Western blots of PrPres were found to be useful indicators of infectivity levels, which at all tested pressures were significantly reduced as compared with untreated controls: from approximately 10(3) LD(50) per g at 690 MPa to approximately 10(6) LD(50) per g at 1,200 MPa. The application of commercially practical conditions of temperature and pressure could ensure the safety of processed meats from bovine spongiform encephalopathy contamination, and could also be used to study phase transitions of the prion protein from its normal to misfolded state. PMID- 12732725 TI - Integration by design. PMID- 12732726 TI - Sexual selection forms the structure and dynamics of ecological communities. PMID- 12732727 TI - New insights into ciliary function: kidney cysts and photoreceptors. PMID- 12732728 TI - NMR structure of a complex containing the TFIIF subunit RAP74 and the RNA polymerase II carboxyl-terminal domain phosphatase FCP1. AB - FCP1 [transcription factor IIF (TFIIF)-associated carboxyl-terminal domain (CTD) phosphatase] is the only identified phosphatase specific for the phosphorylated CTD of RNA polymerase II (RNAP II). The phosphatase activity of FCP1 is enhanced in the presence of the large subunit of TFIIF (RAP74 in humans). It has been demonstrated that the CTD of RAP74 (cterRAP74; residues 436-517) directly interacts with the highly acidic CTD of FCP1 (cterFCP; residues 879-961 in human). In this manuscript, we have determined a high-resolution solution structure of a cterRAP74cterFCP complex by NMR spectroscopy. Interestingly, the cterFCP protein is completely disordered in the unbound state, but forms an alpha helix (H1'; E945-M961) in the complex. The cterRAP74cterFCP binding interface relies extensively on van der Waals contacts between hydrophobic residues from the H2 and H3 helices of cterRAP74 and hydrophobic residues from the H1' helix of cterFCP. The binding interface also contains two critical electrostatic interactions involving aspartic acid residues from H1' of cterFCP and lysine residues from both H2 and H3 of cterRAP74. There are also three additional polar interactions involving highly conserved acidic residues from the H1' helix. The cterRAP74cterFCP complex is the first high-resolution structure between an acidic residue-rich domain from a holoenzyme-associated regulatory protein and a general transcription factor. The structure defines a clear role for both hydrophobic and acidic residues in proteinprotein complexes involving acidic residue-rich domains in transcription regulatory proteins. PMID- 12732729 TI - Polo-like kinase (Plk)1 depletion induces apoptosis in cancer cells. AB - Elevated expression of mammalian polo-like kinase (Plk)1 occurs in many different types of cancers, and Plk1 has been proposed as a novel diagnostic marker for several tumors. We used the recently developed vector-based small interfering RNA technique to specifically deplete Plk1 in cancer cells. We found that Plk1 depletion dramatically inhibited cell proliferation, decreased viability, and resulted in cell-cycle arrest with 4 N DNA content. The formation of dumbbell like chromatin structure suggests the inability of these cells to completely separate the sister chromatids at the onset of anaphase. Plk1 depletion induced apoptosis, as indicated by the appearance of subgenomic DNA in fluorescence activated cell-sorter (FACS) profiles, the activation of caspase 3, and the formation of fragmented nuclei. Plk1-depletion-induced apoptosis was partially reversed by cotransfection of nondegradable mouse Plk1 constructs. In addition, the p53 pathway was shown to be involved in Plk1-depletion-induced apoptosis. DNA damage occurred in Plk1-depleted cells and inhibition of ATM strongly potentiated the lethality of Plk1 depletion. Although p53 is stabilized in Plk1-depleted cells, DNA damage also occurs in p53(-/-) cells. These data support the notion that disruption of Plk1 function could be an important application in cancer therapy. PMID- 12732730 TI - Amyloid-beta peptide levels in brain are inversely correlated with insulysin activity levels in vivo. AB - Factors that elevate amyloid-beta (Abeta) peptide levels are associated with an increased risk for Alzheimer's disease. Insulysin has been identified as one of several proteases potentially involved in Abeta degradation based on its hydrolysis of Abeta peptides in vitro. In this study, in vivo levels of brain Abeta40 and Abeta42 peptides were found to be increased significantly (1.6- and 1.4-fold, respectively) in an insulysin-deficient gene-trap mouse model. A 6-fold increase in the level of the gamma-secretase-generated C-terminal fragment of the Abeta precursor protein in the insulysin-deficient mouse also was found. In mice heterozygous for the insulysin gene trap, in which insulysin activity levels were decreased approximately 50%, brain Abeta peptides were increased to levels intermediate between those in wild-type mice and homozygous insulysin gene-trap mice that had no detectable insulysin activity. These findings indicate that there is an inverse correlation between in vivo insulysin activity levels and brain Abeta peptide levels and suggest that modulation of insulysin activity may alter the risk for Alzheimer's disease. PMID- 12732731 TI - Prevalence of defective DNA mismatch repair and MSH6 mutation in an unselected series of endometrial cancers. AB - Endometrial cancer is the most common gynecologic malignancy in the United States and the most frequent extracolonic tumor in hereditary nonpolyposis colorectal cancer (HNPCC). HNPCC patients have inherited defects in DNA mismatch repair and the microsatellite instability (MSI) tumor phenotype. Sporadic endometrial cancers also exhibit MSI, usually associated with methylation of the MLH1 promoter. Germ-line MSH6 mutations, which are rare in HNPCC, have been reported in several families with multiple members affected with endometrial carcinoma. We reasoned that MSH6 mutation might account for loss of mismatch repair in MSI positive endometrial cancers in which the cause of MSI is unknown. We therefore investigated MSI and MLH1 promoter methylation in 441 endometrial cancer patients unselected for age or personal and family history of cancers. MSI and MLH1 promoter methylation status were associated with age of onset and tumor histology. One hundred cases (23% of the entire series) were evaluated for MSH6 defects. Inactivating germ-line MSH6 mutations were identified in seven women with MSI-positive, MLH1 promoter unmethylated cancers. Most of the MSI in these cases was seen with mononucleotide repeat markers. The MSH6 mutation carriers were significantly younger than the rest of the population (mean age 54.8 versus 64.6, P = 0.04). Somatic mutations were seen in 17 tumors, all of which had MSI. Our data suggest that inherited defects in MSH6 in women with endometrial cancer are relatively common. The minimum estimate of the prevalence of inherited MSH6 mutation in endometrial cancer is 1.6% (7 of 441), comparable with the predicted prevalence for patients with colorectal cancer. PMID- 12732732 TI - Postinsult treatment with lithium reduces brain damage and facilitates neurological recovery in a rat ischemia/reperfusion model. AB - Lithium has long been a primary drug used to treat bipolar mood disorder, even though the drug's therapeutic mechanisms remain obscure. Recent studies demonstrate that lithium has neuroprotective effects against glutamate-induced excitotoxicity in cultured neurons and in vivo. The present study was undertaken to examine whether postinsult treatment with lithium reduces brain damage induced by cerebral ischemia. We found that s.c. injection of lithium dose dependently (0.5-3 mEq/kg) reduced infarct volume in the rat model of middle cerebral artery occlusionreperfusion. Infarct volume was reduced at a therapeutic dose of 1 mEq/kg even when administered up to 3 h after the onset of ischemia. Neurological deficits induced by ischemia were also reduced by daily administration of lithium over 1 week. Moreover, lithium treatment decreased the number of neurons showing DNA damage in the ischemic brain. These neuroprotective effects were associated with an up-regulation of cytoprotective heat shock protein 70 (HSP70) in the ischemic brain hemisphere as determined by immunohistochemistry and Western blotting analysis. Lithium-induced HSP70 up-regulation in the ischemic hemisphere was preceded by an increase in the DNA binding activity of heat shock factor 1, which regulates the transcription of HSP70. Physical variables and cerebral blood flow were unchanged by lithium treatment. Our results suggest that postinsult lithium treatment reduces both ischemia-induced brain damage and associated neurological deficits. Moreover, the heat shock response is likely to be involved in lithium's neuroprotective actions. Additionally, our studies indicate that lithium may have clinical utility for the treatment of patients with acute stroke. PMID- 12732733 TI - Binding and recognition in the assembly of an active BRCA1/BARD1 ubiquitin-ligase complex. AB - BRCA1 is a breast and ovarian cancer tumor suppressor protein that associates with BARD1 to form a RINGRING heterodimer. The BRCA1BARD1 RING complex functions as an ubiquitin (Ub) ligase with activity substantially greater than individual BRCA1 or BARD1 subunits. By using NMR spectroscopy and site-directed mutagenesis, we have mapped the binding site on the BRCA1BARD1 heterodimer for the Ub conjugating enzyme UbcH5c. The results demonstrate that UbcH5c binds only to the BRCA1 RING domain and not the BARD1 RING. The binding interface is formed by the first and second Zn(2+)-loops and central alpha-helix of the BRCA1 RING domain, a region disrupted by cancer-predisposing mutations. Unexpectedly, a second Ub conjugating enzyme, UbcH7, also interacts with the BRCA1BARD1 complex with similar affinity, although it is not active in Ub-ligase activity assays. Thus, binding alone is not sufficient for BRCA1-dependent Ub-ligase activity. PMID- 12732735 TI - Materials become insensitive to flaws at nanoscale: lessons from nature. AB - Natural materials such as bone, tooth, and nacre are nanocomposites of proteins and minerals with superior strength. Why is the nanometer scale so important to such materials? Can we learn from this to produce superior nanomaterials in the laboratory? These questions motivate the present study where we show that the nanocomposites in nature exhibit a generic mechanical structure in which the nanometer size of mineral particles is selected to ensure optimum strength and maximum tolerance of flaws (robustness). We further show that the widely used engineering concept of stress concentration at flaws is no longer valid for nanomaterial design. PMID- 12732737 TI - An international perspective on wound pain and trauma. PMID- 12732734 TI - The structure of nonvertebrate actin: implications for the ATP hydrolytic mechanism. AB - The structures of Saccharomyces cerevisiae, Dictyostelium, and Caenorhabditis elegans actin bound to gelsolin segment-1 have been solved and refined at resolutions between 1.9 and 1.75 A. These structures reveal several features relevant to the ATP hydrolytic mechanism, including identification of the nucleophilic water and the roles of Gln-137 and His-161 in positioning and activating the catalytic water, respectively. The involvement of these residues in the catalytic mechanism is consistent with yeast genetics studies. This work highlights both structural and mechanistic similarities with the small and trimeric G proteins and restricts the types of mechanisms responsible for the considerable enhancement of ATP hydrolysis associated with actin polymerization. The conservation of functionalities involved in nucleotide binding and catalysis also provide insights into the mechanistic features of members of the family of actin-related proteins. PMID- 12732738 TI - Stomal complications: at what price? PMID- 12732739 TI - Treating recalcitrant diabetic wounds with hyaluronic acid: a review of patients. AB - The wound healing process in the patient with diabetes can be adversely affected by many factors, including unrelieved pressure, infection, and concurrent underlying conditions. In addition, a static wound may exacerbate patient anxiety or depression and indirectly further delay the healing process while increasing the risk of complications. The polysaccharide hyaluronic acid has long been known to be an integral component of the extracellular matrix in the dermis and other tissues and is implicated in the process of wound healing and tissue repair. Research has shown the benefit of using a novel ester of hyaluronic acid to accelerate the healing process and effectively treat diabetic foot ulceration and other difficult-to-heal chronic wounds. The effects of care using hyaluronic acid on three patients with challenging wounds are presented. PMID- 12732740 TI - Pharmacotherapy as adjunctive treatment for serious foot wounds in the patient with diabetes: a case study. AB - Chronic foot wounds in patients with diabetes present significant treatment challenges. A 54-year-old woman with type 2 diabetes and two wounds (one on the left great toe and the other on the left medial plantar surface) visited the clinic after the chronic wounds failed to respond to treatment such as hydrotherapy. Subsequent comprehensive care, including debridement; opening tunnels distal, proximal, and medial from the plantar wound; application of a growth factor-stimulant; and treatment with cilostazol to improve both macro- and microvascular circulation provided excellent wound healing. Amputation was avoided and the patient returned to her regular routine within 6 months. Pharmacotherapy may provide new adjunctive therapy options in the treatment of chronic foot wounds in patients with diabetes mellitus. Controlled clinical studies to ascertain the effects of this treatment are warranted. PMID- 12732741 TI - Does sterile or nonsterile technique make a difference in wounds healing by secondary intention? AB - After observing inconsistencies in care of acute surgical wounds healing by secondary intention and reviewing the potential cost savings of implementing clean dressing change technique policies, surgical nurses at a university-based medical center monitored supply usage and infection rates of these wounds using a nonexperimental, longitudinal study design. Staff from two acute care surgical units provided data for 3 months before and 3 months after standardization of wound care to a clean wound care technique. All adult patients requiring dressing changes three times per day with normal saline moistened gauze of their open surgical wound(s) participated in the study. Before changing the wound care procedures, nine (9) of 1,070 (0.84%) admissions to the two surgical units had a surgical site infection. During the 3 months following implementation of clean wound care protocols, eight (8) surgical site infections were documented in 963 admissions (rate.83%). Dressing supply costs were $380 less. In this study, using nonsterile wound care procedures for wounds healing by secondary intention did not negatively impact infection rates and saved supply costs. PMID- 12732742 TI - Epidermoid cancers that masquerade as venous ulcer disease. AB - Many lesions originally diagnosed as venous ulcers exhibit characteristics that are strikingly similar to skin cancers and might represent sites of primary carcinomas. To ascertain the frequency of malignancy in patients previously diagnosed with venous ulcer disease, a retrospective cohort review of patients evaluated at a Wound Healing Center in Florida was conducted. Charts of all patients with IDC-9 codes for varicose veins with stasis ulcer, varicose veins with ulcer and inflammation, and venous peripheral insufficiency were reviewed. Only charts of patients with one of these diagnoses and documented clinical varicosities, hemosiderosis, brawny edema, and lesions located at the medial or lateral lower leg were included. Sixty (60) patients were identified. Of these, 20 had lesions that were clinically suspicious for epidermoid skin cancers (ie, showing raised borders and chronic scaling). Biopsies confirmed malignancy in 15 of the 60 ulcers (25%). Of these, eight were squamous cell cancers. Given the high rate of malignancies in this cohort of patients, it is postulated that primary epidermoid cancers may mimic venous ulcers in appearance, location, and symptoms; that Marjolin's ulcers are rare despite their propensity to develop in many different types of wounds; and that patients with a history of venous ulcers and prolonged exposure to ultraviolet rays may benefit from lesion biopsies to test for epidermoid cancers. PMID- 12732743 TI - The voice of great events. PMID- 12732744 TI - The human element of advanced technology. AB - Human beings are uniquely endowed with the power and desire to manipulate the world around them. Although this drive has been present throughout the ages, today vast sources of knowledge provide a means to create products of advanced technology unlike any before in recorded history. Humankind is driven to control the human condition ie, the inherent tragedies of life. Yet, from their inception, human technological creations have the potential to lead to tragic ends. Some contend this is because of the ever-present human element within the products of advanced technology. The human factor inherent in advanced technology is reviewed with attention to the risks and benefits therein. Current case studies are presented. The ethical principle of informed consent is presented within the context of risks and benefits of advanced technology. PMID- 12732745 TI - Black and white. PMID- 12732746 TI - Pain doesn't have to be a part of wound care. PMID- 12732747 TI - The hype in hypoallergenic. PMID- 12732749 TI - Calciphylaxis: a case study with differential diagnosis. AB - Calciphylaxis is a relatively rare, but horribly disfiguring, skin condition that is most often associated with end-stage renal disease and long-term dialysis. Unfortunately, calciphylaxis-related morbidity and mortality are significant. The case study presented demonstrates many of the findings associated with the typical calciphylaxis patient; end-stage renal disease and an extensive, painful ulcer. The complexity of the patient s history and medical/surgical interventions, especially medication therapy with coumadin and heparin, complicated initial diagnostic processes. Close scrutiny of multiple physical assessment findings, historical factors, and test results was required for correct diagnosis. Crucial components of differential diagnosis of calciphylaxis versus coumadin-induced skin necrosis or heparin-induced thrombocytopenia necrosis include: patient history and characteristics, clinical presentation, and diagnostic test results. PMID- 12732750 TI - Analysis of localized erythema using clinical indicators and spectroscopy. AB - Localized erythema is regularly used as an indicator of post-ischemic events, including reactive hyperemia and Stage I pressure ulcers. The National Pressure Ulcer Advisory Panel definition of a Stage I ulcer includes both visual and nonvisual indicators in part to improve identification in darkly pigmented skin. A prospective, repeated-measures design was used to collect information on pressure-induced erythema that includes reactive hyperemia and Stage I pressure ulcers with an emphasis on distinguishing indicators in light and dark skin The relationships among clinical indicators (skin assessments) and results from tissue reflectance spectroscopy, as well as the clinical utility of spectroscopy for discerning tissue blanching status, were examined in a convenience sample of 76 inpatients and outpatients (95 test/control site pairs). Chi-square analysis and generalized logistic models were used to identify relationships and distinguishing characteristics of erythema. Analysis of variance was used to analyze blanching using spectroscopy. Nonblanching sites were more likely to be persistent erythema (c2=5.3; P = 0.021) but exhibited no relationships to temperature, tissue resilience, or disability. Erythema in subjects with dark skin was more likely to be nonblanching and have poor resilience. Spectrographic analysis of blanching found significant differences across skin pigmentation (P = 0.0001) and blanching status (P = 0.019). These results reinforce the belief that dark skin must be assessed differently than light skin and indicate that clinicians should use persistence of erythema rather than blanching status to judge incipient pressure ulcers. These results validate the use of visual and nonvisual indicators included in the National Pressure Ulcer Advisory Panel Stage I pressure ulcer definition. PMID- 12732751 TI - Knowledge of pressure ulcers by undergraduate nursing students in Brazil. AB - The purpose of this study was to examine Brazilian nursing students knowledge of pressure ulcers. Third- and fourth-year undergraduate baccalaureate students at a public university in Brazil (N = 83) were asked to provide demographic information, identify extracurricular activities (eg, reading journals and articles and using the Internet to enhance comprehension of pressure ulcer care), and complete the Pressure Ulcer Knowledge Test. Students correctly answered 67.7% of the Pressure Ulcer Knowledge Test items. Students who participated in extracurricular activities and used the Internet had significantly higher Knowledge Test scores than those who did not. Readings did not significantly impact the Knowledge Test score. Generally, the students were found to have low pressure ulcer knowledge, but that educational programs and the Internet have the potential to positively impact nursing students knowledge of pressure ulcers. PMID- 12732752 TI - Pressure ulcer prevention: where did we go wrong? PMID- 12732753 TI - Medicaid: for want of an ostomy pouch. PMID- 12732754 TI - A pain management protocol for wound care. PMID- 12732755 TI - The evolution and innovation of ostomy products. PMID- 12732757 TI - Determining the efficacy of a pressure ulcer prevention program by collecting prevalence and incidence data: a unit-based effort. AB - Pressure ulcer prevention falls within the domain of nursing practice. When the results of a quality improvement survey indicated both an increase in the number of pressure ulcers and a higher prevalence than the national average, the nursing staff of a 500-bed Midwest hospital developed a pressure ulcer prevention program guided by the AHCPR guidelines. The literature supports collecting prevalence and incidence data as indicators of prevention program effectiveness, and the best indicator of the effectiveness of prevention strategies to reduce nosocomial pressure ulcers is incidence. Since the tracking mechanism was instituted, awareness of the results and impact of prevention measures increased; most nursing units experienced a 10% to 20% decrease in the incidence of pressure ulcers. Designing an efficient, timely, and practical method of retrieving pressure ulcer prevalence and incidence data provided a quality assurance method of monitoring the success of the program. PMID- 12732756 TI - Assessing venous ulcer population characteristics and practices in a home care community. AB - To plan for a new community leg ulcer service in one Ontario region, venous ulcer population characteristics and current community care practices were obtained from a home care cohort of people with venous ulcers. A secondary analysis was conducted on the venous ulcer cohort using data collected during a Regional Prevalence and Profile Study. Patients were identified based on the clinical syndrome for venous disease criteria described in the Royal College of Nurses clinical practice guidelines and by the presence of an ankle-brachial pressure index 0.8. The Regional Prevalence and Profile Study identified 263 people with leg ulcers for a rate of 2.0 per 1,000 people >25 years of age. One hundred, seven ulcers (41%) were the result of venous disease; of these, 83 (78% of cases) were associated with a single nursing agency and formed the study cohort. Most patients (51, 61%) were female and 65 years old. Thirty-eight (46%) had 4 comorbid conditions, 63 spoke English, 29 lived alone, 38 did not require physical aids or assistance for mobility, and 81 (98%) were able to travel outside of their home. The current ulcer had been present for an average of 15 months (median 6 months), 51 participants had a previous leg ulcer, and 22 had episodes of ulceration for > 5 years. Of the 121 ulcers in the study, 48 (41%) were located at the ankle, and the majority (85%) were > 1 cm2. General practitioners were the main medical care providers for 48 participants, and 52 (62%) had seen a specialist physician for their current ulcer. These findings are similar to large studies conducted in other industrialized countries and confirm that venous ulcers are a chronic problem in a population with complex health needs. PMID- 12732758 TI - Wound infections in two health institutions in Ile-Ife, Nigeria: results of a cohort study. AB - The control of wound infections is increasingly complicated, yet treatment is not always guided by microbiological diagnosis. To describe the distribution of wound infections and causative agents, a prospective, 6-month cohort study involving 102 outpatients was conducted at the University Teaching Hospital and the Health Center in Ile-Ife, Nigeria. Location and type of infected wounds were recorded and bacterial isolates were identified by standard microbiological techniques. Almost half (40%) of all infected wounds were attributed to trauma and, in most cases, located on the extremities. Of the 162 bacterial isolates obtained from wound cultures, 39 were monomicrobial and 55 were polymicrobial; no bacterial isolate was obtained in eight cases. Staphylococcus aureus was the predominant micro-organism (25%), followed by Escherichia coli (12%), Pseudomonas aeruginosa (9%), and Staphylococcus epidermidis (9%). The diversity of micro-organisms and the high incidence of polymicrobic flora in this study give credence to the value of identifying one or more bacterial pathogens from wound cultures. The recognition of causative agents of wound infections can assist wound care practitioners in taking appropriate measures. Continuous dialogue between the microbiology department and wound care practitioners is strongly advised in order to improve treatment outcomes and slow the development of antibiotic-resistant bacteria. PMID- 12732759 TI - Laparoscopic-assisted vaginal hysterectomy: American Association of Gynecologic Laparoscopists' 2000 membership survey. PMID- 12732760 TI - Miliary tuberculous peritonitis. PMID- 12732761 TI - Heterotopic pregnancy. PMID- 12732762 TI - Major complications associated with laparoscopic-assisted vaginal hysterectomy: ten-year experience. AB - STUDY OBJECTIVE: To describe our experience with major complications associated with laparoscopic-assisted vaginal hysterectomy (LAVH) and compare our results with those of the American Association of Gynecologic Laparoscopists (AAGL) membership survey and another similar study. DESIGN: Retrospective study (Canadian Task Force classification II-3). SETTING: University-affiliated hospital. PATIENTS: Two thousand seven hundred two women. Intervention. LAVH. MEASUREMENTS AND MAIN RESULTS: Demographic data and medical histories (age, parity, surgical indications, pathologic findings, major complications) were analyzed. Major complications were 11 bladder injuries, 4 ureter injuries, 11 bowel injuries, 2 vascular injuries, 2 cases of massive bleeding from the vaginal cuff or colpotomy wound with associated impending shock, 2 cases of postoperative ileus, and 2 pelvic abscesses. Our overall major complication rate was 1.3% compared with 2.7% in the AAGL 1995 membership survey (p <0.001). Similar rates of febrile morbidity (2.2% and 2.0%), bleeding requiring transfusion (0.05% and 0.06%), and bowel, ureteral, or bladder injury (1.0% and 1.0%) were noted between our study and the other 1995 study (all p >0.05). Of 34 major complications in our study, 24 occurred during hysterectomy performed by inexperienced general gynecologists and 10 by an experienced endoscopist (p = 0.005). CONCLUSION: The rate of major complications associated with LAVH can be reduced when the procedure is performed by a well-trained laparoscopic surgeon compared with a less-experienced general gynecologist. PMID- 12732763 TI - Laparoscopic approach to dermoid cysts: combined surgical technique and ultrasonographic evaluation of residual functioning ovarian tissue. AB - STUDY OBJECTIVE: To estimate how and if laparoscopic removal of ovarian dermoid cysts is a tissue-sparing procedure. DESIGN: Prospective study (Canadian Task Force classification II-2). SETTING: University-associated hospital. PATIENTS: Fifty-five women. INTERVENTION: Laparoscopic removal of ovarian dermoid cysts by a combination of hydrodissection and blunt dissection, and transvaginal sonographic (TVS) evaluation of residual ovarian tissue. MEASUREMENTS AND MAIN RESULTS: Within 1 week before surgery all recruited patients underwent TVS evaluation of ovarian volume, size, and morphology of dermoid cysts and measurement of surrounding ovarian cortex. Mean cyst diameter was 5.5 +/- 2.2 cm (range 2.1-15.0 cm). Within 6 to 12 months after laparoscopic excision, TVS measurements of residual ovarian tissues were obtained. Ovarian residual cortex surrounding the cyst was not visible at TVS in 24 ovaries, whereas in 56 ovaries residual tissue volume was greater than 3 cm3 after laparoscopic excision. CONCLUSION: We propose laparoscopic removal of dermoid cysts by combining hydrodissection and blunt dissection with maximum tissue sparing, even when the cyst seems to fill the ovary and no surrounding ovarian cortex can be seen on ultrasound. PMID- 12732764 TI - Neuroendocrine stress response in patients undergoing benign ovarian cyst surgery by laparoscopy, minilaparotomy, and laparotomy. AB - STUDY OBJECTIVE: To quantify and compare neuroendocrine stress responses during and immediately after surgery by laparoscopy, minilaparotomy, and laparotomy for benign ovarian cysts. DESIGN: Prospective study (Canadian Task Force classification II-1). SETTING: Tertiary care university hospital. PATIENTS: Thirty healthy women with no major diseases and without endocrine disorders. INTERVENTIONS: Surgery for benign ovarian cysts performed by laparoscopy (10), minilaparotomy (10), or laparotomy (10). MEASUREMENTS AND MAIN RESULTS: Venous blood samples were collected at fixed times as follows: at 8 A.M. in the ward before transferring the patient to the operating room (time 0), 30 minutes after the beginning of surgery (time 1), at the end of surgery after extubation with the patient awake (time 2), and 2 and 4 hours after the end of surgery (times 3 and 4). We evaluated intraoperative and postoperative variations of the following stress-related markers: norepinephrine (NE), epinephrine (E), adrenocorticotropic hormone (ACTH), human growth hormone (hGH), prolactin (PRL), and cortisol, and postoperative pain. No differences were present in demographic characteristics and operating times in the three groups. No anesthesiologic or surgical complications occurred. Postoperative pain was similar in the laparoscopy and minilaparotomy group but significantly higher in the laparotomy group (p <0.001). Serum levels of markers were not significantly different among the groups at baseline. In the laparoscopy group the increase of hGH was limited to intraoperative time (p <0.05); increases in NE, E, ACTH, and PRL were limited to intraoperative and early postoperative time after extubation (p <0.01), with only PRL persisting with significantly higher levels after the end of surgery (p <0.05). In the minilaparotomy group no increase was detected for hGH, a significant intraoperative increase in cortisol was present (p <0.05), and NE, E, ACTH, and PRL were significantly higher even after the end of surgery (p <0.01). In this group levels of NE, E, and hGH were significantly higher than in the laparoscopy group 2 and 4 hours after the end of surgery (p <0.05). In the laparotomy group significant intraoperative increases were present for all stress markers and persisted until after extubation for ACTH (p <0.01) and to the postoperative period for NE (p <0.01), E (p <0.01), cortisol (p <0.01), PRL (p <0.05), and hGH (p <0.01). In this group levels of NE, E, ACTH, and hGH were significantly higher than those in the laparoscopy group from the beginning (NE p <0.05, E p <0.01, ACTH p <0.05, hGH p <0.01) until after the end of surgery. Comparison of laparotomy and minilaparotomy groups showed the former to have significantly higher plasma levels of E, cortisol, and hGH in intraoperative and postoperative times (p <0.001); significantly higher NE at sampling times 1 and 2 (p <0.001) and time 4 (p <0.01), and no difference at sampling time 3; and ACTH significantly higher only during surgery (p <0.01). CONCLUSION: Laparoscopic surgery causes minimal activation of stress hormones, which in some instances is confined to the intraoperative period. Minilaparotomy may be a valid alternative to laparoscopy in high-risk patients who cannot tolerate abdominal distention. PMID- 12732765 TI - Laparoscopic appendectomy. AB - STUDY OBJECTIVE: To evaluate the effectiveness of laparoscopic appendectomy in women with chronic pelvic pain and to identify histopathology of the appendix. DESIGN: Retrospective review over 6.5 years (Canadian Task Force classification II-3). SETTING: Laparoscopic center and community hospital. PATIENTS: Three hundred seventeen women. INTERVENTION: Laparoscopic appendectomy in conjunction with other procedures. MEASUREMENTS AND MAIN RESULTS: Of 317 patients who underwent appendectomy, 14 (4.4%) had involvement of the appendix with endometriosis, 12 (3.78%) had early acute appendicitis, 4 (1.26%) had carcinoid tumors of the appendix, 2 (0.63%) had a large mucocele, and 1 (0.9%) each had Enterobius vermicularis infection, benign neuroma, and mucinous cystadenoma. Seventy-eight women (24.6%) had obliteration of the appendiceal lumen and 22 (6.93%) had entrapping fibrous adhesions. Thirty-two patients (10%) reported relief of chronic pelvic pain in the absence of other pathology just by having diagnostic laparoscopy with appendectomy. CONCLUSION: The appendix is a key organ in the evaluation of undiagnosed chronic pelvic pain. PMID- 12732766 TI - Laparoscopic tubal sterilization after two or more cesarean sections. AB - STUDY OBJECTIVE: To determine the safety of laparoscopic tubal sterilization in women who have had two or more cesarean sections. DESIGN: Retrospective study (Canadian Task Force classification III). SETTING: Private clinic and hospitals. PATIENTS: Two hundred ten consecutive women. INTERVENTION: Laparoscopic tubal sterilization. MEASUREMENTS AND MAIN RESULTS: The procedures were done with no difficulty in all but two women. In addition to the two failures, one woman experienced bladder trauma. CONCLUSION: Laparoscopic sterilization after several cesarean sections is possible and is associated with low morbidity. Scarring from cesarean sections should not be a contraindication if extra care is taken. PMID- 12732767 TI - Chance of adhesion formation after laparoscopic salpingo-ovariolysis: is there a place for second-look laparoscopy? AB - STUDY OBJECTIVE: To evaluate the chance of adhesion formation after laparoscopic salpingo-ovariolysis and determine the efficacy of early second-look laparoscopy (SLL). DESIGN: Prospective, randomized study (Canadian Task Force classification I). SETTING: Shiraz University hospitals. SUBJECT: Ninety women with mean duration of infertility of 7.2 years. INTERVENTIONS: Operative laparoscopy, with early SLL with adhesiolysis in 46 (group 1) and no SLL in 44 women (group 2). MEASUREMENTS AND MAIN RESULTS: Adnexal adhesions were evaluated according to American Society for Reproductive Medicine adhesion classification. Separation of newly reformed adhesions was performed at the time of SLL. Patients were followed for a year after operation without other infertility treatment. At the time of operation in group 1, adnexal adhesions were graded as severe (class D) in 19 women, moderate (class C) in 31, mild (class B) in 28, and minimal (class A) in 14. Respective figures in group 2 were 10, 30, 34, and 14. After salpingo ovariolysis these figures were 12, 10, 20, and 50 in group 1 and 6, 14, 17, and 51 in group 2. In group 1 in whom early second-look laparoscopy was performed, at the start of the operation these figures were 17, 20, 21, and 34, and after operation 12, 8, 20, and 52, respectively. There were 11 term pregnancies in group 1 and 15 in group 2. No women with severe adhesions in either group conceived. In group 1, chances of term pregnancy were 18.75% for those with moderate adhesions, 35.71% for women with mild adhesions, and 42.86% in patients with minimal adhesions. Respective figures in group 2 were 26.67%, 41.18%, and 57.14%. CONCLUSION: The chance of moderate and severe adhesion reformation after laparoscopic salpingo-ovariolysis was 40.2%. Although separation of these adhesions could be performed more easily at the time of early SLL, the chance of pregnancy did not increase compared with that in patients who did not undergo SLL. PMID- 12732768 TI - Pregnancy outcomes and deliveries after laparoscopic myomectomy. AB - STUDY OBJECTIVE: To assess pregnancy outcomes and deliveries after laparoscopic myomectomy. DESIGN: Retrospective study (Canadian Task Force classification II 2). SETTING: General hospital. PATIENTS: Three hundred fifty-nine women. INTERVENTIONS: Laparoscopic myomectomy and laparoscopic and/or hysteroscopic treatment of associated pathologies. MEASUREMENTS AND MAIN RESULTS: Five patients (1.39%) were lost to follow-up. Seventy-two women were pregnant at least once after laparoscopic myomectomy, for a total of 76 pregnancies. Four women conceived twice and four are pregnant as of this writing. One multiple pregnancy occurred. Twelve pregnancies resulted in first-trimester miscarriage, one in an ectopic pregnancy, one in a blighted ovum, and one in a hydatiform mole. One patient underwent elective first-trimester termination of pregnancy. Thirty-one women had vaginal delivery at term and 26 were delivered by cesarean section. No case of uterine rupture or dehiscence occurred. CONCLUSION: Our technique of laparoscopic myomectomy appears to allow safe vaginal delivery. PMID- 12732769 TI - Rectal surgery for endometriosis--should we be aggressive? AB - STUDY OBJECTIVE: To assess the outcome of aggressive but conservative laparoscopic surgery in the treatment of severe endometriosis involving the rectum. DESIGN: Retrospective study (Canadian Task Force classification III). SETTING: Endosurgery unit of a tertiary referral center. PATIENTS: One hundred sixty-nine women. INTERVENTION: Laparoscopy or laparotomy. MEASUREMENTS AND MAIN RESULTS: The procedure was completed successfully laparoscopically in 145 (86%) and by laparotomy in 24 women (14%). The rate of preoperative symptoms was higher in 25 women who underwent bowel resection compared with those who had other bowel surgery. In addition to bowel surgery, excision of uterosacral ligaments, adhesiolysis, excision of endometrioma, and oophorectomy were the four most commonly performed procedures. At 35-month follow-up 61 patients (36%) required further surgery for pain. The average time between primary and repeat surgery was 16 months. This second operation was performed by laparoscopy in over three fourths of the women. Overall recurrent endometriosis was found in 26 patients (15%). Overall morbidity associated with all surgery was 12.4%. CONCLUSION: Surgery for endometriosis of the cul-de-sac and bowel involves some of the most difficult dissections encountered, but it can be accomplished successfully with the low postoperative morbidity typical of laparoscopy. PMID- 12732770 TI - Laparoscopic mobilization of the rectosigmoid and excision of the obliterated cul de-sac. AB - STUDY OBJECTIVE: To evaluate the feasibility and surgical and clinical outcomes of laparoscopic excision of anterior recto-sigmoid wall endometriosis and en bloc dissection of the obliterated cul-de-sac. DESIGN: Retrospective cohort (Canadian Task Force classification II-2). SETTING: University-affiliated teaching hospital. PATIENTS: Eighty-one women with infertility and/or chronic pelvic pain. Intervention. Laparoscopic excision of all endometrial implants and uterosacral ligaments, and dissection of the cul-de-sac using a horseshoe-shaped approach to mobilize, but not resect, the rectosigmoid. MEASUREMENTS AND MAIN OUTCOMES: Eleven women (24%) had endometriomas. Cumulative pregnancy rates in 34 women with primary infertility and 12 with secondary infertility were 62% and 42%, respectively. Eighty-eight percent of 61 women with pain reported significant improvement of symptoms. CONCLUSION: Laparoscopic excision of cul-de-sac and rectovaginal endometriosis by this approach is feasible and safe when performed by an experienced surgeon, and results in high rates of cumulative pregnancy and relief of pain. Some patient variables may give higher rates of success for pregnancy than others. PMID- 12732771 TI - Total laparoscopic surgery of cystic adenomyoma under hydroultrasonographic monitoring. AB - A 19-year-old-woman had a cystic adenomyoma located within the myometrium. She complained of severe dysmenorrhea. Gonadotropin-releasing hormone agonist therapy was administered, but her dysmenorrhea was more pronounced than before treatment. Therefore, total laparoscopic resection of the lesion was performed. The external appearance of the patient's uterus was almost normal. A hydroultrasonographic monitoring method was devised (transvaginal ultrasonography with peritoneal hydration of physiologic saline) that proved useful in locating the adenomyoma. The patient's dysmenorrhea disappeared postoperatively. PMID- 12732773 TI - Chemically assisted dissection of tissues in laparoscopic excision of endometriotic cysts. AB - STUDY OBJECTIVE: To evaluate the capacity of chemical dissection of tissues using a mucolytic substance, Mesna, in improving laparoscopic excision of endometriotic cysts. DESIGN: Randomized, double-blind, controlled trial (Canadian Task Force classification I). SETTING: University-affiliated training hospital. PATIENTS: Forty-four women with symptomatic ovarian endometriotic cysts. Intervention. Laparoscopic excision of endometriotic cysts in 22 women with the aid of Mesna solution and in 22 with the aid of saline solution. MEASUREMENTS AND MAIN RESULTS: In comparison with saline solution, Mesna as a chemical dissector resulted in significant reductions in operating time, in difficulty encountered by the surgeon to enucleate the cysts, and in less bleeding. No differences were found in length of hospital stay, costs of surgeries, analgesic requirement, and fever. Postoperatively, patients treated with Mesna achieved more pregnancies than those treated with saline. CONCLUSION: Chemical dissection of tissues with Mesna proved to be a safe and suitable support in laparoscopic surgery for ovarian endometriotic cysts. PMID- 12732772 TI - Pregnancy outcome after laparoscopy or laparotomy in pregnancy. AB - STUDY OBJECTIVE: To assess obstetric performance and fetal outcomes after laparoscopy or laparotomy performed during pregnancy. DESIGN: Nationwide, multicenter, retrospective chart review (Canadian Task Force classification II 2). SETTING: Seventeen hospitals throughout Israel: 12 university or university affiliated hospitals and 5 general hospitals. PATIENTS: Three hundred eighty-nine pregnant women. INTERVENTION: Laparoscopy or laparotomy for various indications. MEASUREMENTS AND MAIN RESULTS: Of 192 laparoscopies performed, 141 were during the first, 46 during the second, and 5 during the third trimester; respective figures for 197 laparotomies were 63, 110, and 24. No intraoperative complications were reported for either procedure. Six and 25 women had complications after laparoscopy and laparotomy, respectively. There was no significant difference in abortion rates between groups. Mean gestational age at delivery and mean birthweight were comparable between groups. No significant difference was found in frequency of fetal anomalies between groups or when compared with the Israel register of anomalies. CONCLUSION: Operative laparoscopy seems to be as safe as laparotomy in pregnancy. PMID- 12732774 TI - Effects of closed suction drainage in reducing pain after laparoscopic-assisted vaginal hysterectomy. AB - STUDY OBJECTIVE: To estimate whether closed suction drainage of the pelvis after laparoscopic-assisted vaginal hysterectomy (LAVH) reduces the frequency and intensity of shoulder-tip, abdominal, and back pain. DESIGN: Prospective, randomized study (Canadian Task Force classification 1). SETTING: Teaching medical center. PATIENTS: One hundred sixty-four women. INTERVENTION: LAVH. MEASUREMENTS AND MAIN RESULTS: For group 1 (80 women), closed suction (Jackson Pratt) drains were inserted into the peritoneal cavity and cul-de-sac, whereas for group 2 (84), no drains were placed. Shoulder-tip, abdominal, and back pain were evaluated by visual analog scores (VAS) 3, 24, and 48 hours after surgery. The frequency of shoulder-tip pain was significantly lower in group 1 at 24 hours (23% vs 40%, p = 0.013) and 48 hours (9% vs 21%, p = 0.024; VAS scores at 24 hrs 2.2 +/- 1.1 vs 3.8 +/- 1.3, p = 0.010; VAS scores at 48 hours 1.5 +/- 1.0 vs 2.5 +/- 1.2, p = 0.018). At 48 hours fewer women in group 1 experienced abdominal pain (31% vs 50%, p = 0.039; VAS scores 2.0 +/- 1.1 vs 4.0 +/- 1.3, p = 0.007). No statistically significant differences in frequency and VAS scores for back pain were found at any time. The quantity of oral analgesics was greater for group 2 than for group 1 (12.4 +/- 1.6 vs 9.0 +/- 1.4, p <0.001). Economic evaluation of analgesic requirements and material costs for the two groups showed that simple analgesics were more cost-effective than closed suction drainage in the treatment of pain. CONCLUSION: Closed suction drains may reduce the frequency and intensity of shoulder-tip and abdominal pain and postoperative analgesia requirements after LAVH, but simple oral analgesics are more cost effective. PMID- 12732775 TI - Combination of laparoscopic bilateral uterine artery ligation and intraamniotic methotrexate injection for conservative management of cervical pregnancy. AB - STUDY OBJECTIVE: To evaluate whether the combination of laparoscopic bilateral uterine artery ligation and intraamniotic methotrexate injection may eliminate unexpected and uncontrolled massive uterine bleeding without compromising future fertility in women with cervical pregnancy. DESIGN: Prospective study (Canadian Task Force classification II-2). SETTING: Tertiary-care university hospital. PATIENTS: Three women. INTERVENTION: Laparoscopic bilateral uterine artery ligation and intraamniotic methotrexate injection. MEASUREMENTS AND MAIN RESULTS: Three cases of cervical pregnancy were diagnosed by ultrasound at 6, 7, and 9 weeks' gestation. After treatment, all three women experienced intermittent vaginal bleeding, but none required transfusion. Levels of b-human chorionic gonadotropin returned to normal within 7 weeks, and patients resumed normal menstruation within 11 weeks after treatment. One woman conceived an intrauterine pregnancy 3 months after restoration of normal menstruation, and was delivered at term. CONCLUSION: The combination of laparoscopic bilateral uterine artery ligation and intraamniotic methotrexate injection appears to be effective in preventing unexpected massive uterine bleeding in patients with cervical pregnancy, and does not compromise future fertility. PMID- 12732776 TI - Transvaginal hydrolaparoscopic ovarian drilling using bipolar electrosurgery to treat anovulatory women with polycystic ovary syndrome. AB - STUDY OBJECTIVE: To verify the value, feasibility, and reliability of transvaginal hydrolaparoscopic ovarian drilling using the bipolar VersaPoint system to treat clomiphene-resistant, anovulatory women with polycystic ovary syndrome. DESIGN: Prospective clinical study (Canadian Task Force classification II-2). SETTING: University teaching hospital. PATIENTS: Twenty-eight women. INTERVENTION: Transvaginal hydrolaparoscopic ovarian drilling using the bipolar VersaPoint spring electrode. MEASUREMENTS AND MAIN RESULTS: After the surgical procedure, ovulation occurred spontaneously in 66.7% of women. Thirteen pregnancies occurred; eight were spontaneous, and four were achieved after induction of ovulation with clomiphene citrate and one after stimulation with gonadotropins. In 21 women whose infertility was due exclusively to anovulation, the cumulative pregnancy rate was 38% at 3 months and 76% at 6 months. No ovarian hyperstimulation or abortion occurred. The single complication was bleeding from an ovary that required conversion to laparoscopy. CONCLUSION: Transvaginal hydrolaparoscopic ovarian drilling with the bipolar VersaPoint system is a useful therapeutic option in these women. PMID- 12732777 TI - Assessment and comparison of intraoperative and postoperative pain associated with NovaSure and ThermaChoice endometrial ablation systems. AB - STUDY OBJECTIVE: To assess and compare intraoperative and postoperative pain associated with NovaSure impedance-controlled endometrial ablation system and ThermaChoice system. DESIGN: Prospective, international, multicenter, double-arm study (Canadian Task Force classification II-1). Setting. Academic medical centers and private offices. PATIENTS: Sixty-seven premenopausal women with menorrhagia. INTERVENTION: Endometrial ablation with either the NovaSure (37 women) or ThermaChoice (30) system. NovaSure-treated patients received no endometrial pretreatment; those treated with ThermaChoice received the recommended 3-minute suction dilatation and curettage. MEASUREMENTS AND MAIN RESULTS: Standard pain measurement instruments (visual analog scale, numeric rating scale) were used to assess intraoperative and postoperative pain. Serum levels of prostaglandin-F(2alpha) were measured before and 5, 30, and 60 minutes after the procedure. Patients treated with the NovaSure system reported statistically significantly lower intraoperative and postoperative pain than those treated with the ThermaChoice system (p <0.0001). Procedure time was statistically significantly shorter with the NovaSure system (p <0.0001). Prostaglandin-F(2alpha) values did not differ statistically between groups. CONCLUSION: The NovaSure system is associated with statistically significantly lower intraoperative and postoperative pain than ThermaChoice system, and endometrial ablation with NovaSure could become an office-based procedure. PMID- 12732779 TI - Repeat hysteroscopic surgery reduces the hysterectomy rate after endometrial and myoma resection. AB - STUDY OBJECTIVE: To assess the efficacy of repeat transcervical resection of the endometrium (TCRE) in patients with dysfunctional uterine bleeding and myomas in whom primary resection failed. DESIGN: Retrospective analysis (Canadian Task Force classification III). SETTING: University hospital endoscopic unit. PATIENTS: Six hundred sixty-eight women. INTERVENTION: Repeat TCRE or transcervical resection of a myoma (TCRM). MEASUREMENTS AND MAIN RESULTS: Of 668 patients, 118 (17%) required repeat resection for the following reasons: pain (52, 44%), menorrhagia (39, 31%), myomas (15, 13%), perforation at the primary TCRE (6, 5%), and large fluid deficit during the procedure (6, 5%). Of 118 women undergoing repeat TCRE or TCRM, 33 (28%) eventually required hysterectomy due to pain (17, 48%), persistent bleeding (7, 27%), pain and bleeding (3, 10%), regrowth of myomas (3, 14%), and other reasons (3, 14%). CONCLUSION: Repeat resection is an option after failed primary hysteroscopic operation and may reduce the hysterectomy rate. PMID- 12732778 TI - Classification of microhysteroscopic images and their correlation with histologic diagnoses. AB - STUDY OBJECTIVE: To confirm that images observed at hysteroscopy correlate with histopathologic diagnoses. DESIGN: Double-blind study (Canadian Task Force classification II-2). SETTING: Gynecologic cancer center, private institute. SUBJECTS: One thousand four hundred thirty-six uterine cavities. INTERVENTION: Hysteroscopy. MEASUREMENTS AND MAIN RESULTS: Images were classified as normal hysteroscopy, benign lesion, low-risk hyperplasia, high-risk hyperplasia, and carcinoma. A hysteroscopic diagnosis was made and biopsy specimens were obtained with Kevorkian-type curettes or Sims curettes. Tissues were studied by a pathologist, after which we compared endoscopic diagnoses with anatopathologic diagnoses. Significant correlation was found between suspicion based on images and histologic confirmation (p = 0.001). The Cramer V coefficient, which measures the relationship between both methods, was high: 0.925. The Cramer V coefficient takes values ranging from zero, to indicate lack of correlation, to 1, to indicate perfect correlation. A lambda symmetry coefficient of 0.96 is interpreted as probable improvement in the prediction of histologic diagnosis based on images. CONCLUSION: This classification system can be useful for a systematic approach to hysteroscopic findings and to improve communication among specialists. It is based on the degree of hysteroscopic suspicion aimed at early detection of endometrial cancer and its precursor lesions. PMID- 12732781 TI - Intrauterine pathology in women with abnormal uterine bleeding taking hormone replacement therapy. AB - STUDY OBJECTIVE: To assess the frequency of intrauterine pathology among women with abnormal uterine bleeding (AUB) while taking hormone replacement therapy (HRT), and correlate pathology with clinical features. DESIGN: Retrospective review (Canadian Task Force classification II-2). SETTING: University teaching hospital-based outpatient clinic. PATIENTS: Ninety-nine women. INTERVENTION: Observational evaluation. MEASUREMENTS AND MAIN RESULTS: Intrauterine pathology was found in 16 (18.6%) of 99 women with AUB. The frequency was 4 times higher in those with AUB after achieving amenorrhea with a continuous combined regimen of HRT than in those with AUB with only 6 months of HRT. It was also higher in patients taking a continuous combined regimen in whom AUB lasted for more than 6 months. CONCLUSION: Hysteroscopic examination with endometrial sampling is recommended if AUB continues after the first 6 months of HRT or if it occurs after a period of amenorrhea. PMID- 12732782 TI - A new technique of hysteroscopic myomectomy with enucleation in toto. AB - STUDY OBJECTIVE: To determine the safety and effectiveness of a new technique for hysteroscopic resection of uterine submucous myomas with high intramural involvement (G2 type, European Society of Hysteroscopy classification). DESIGN: Prospective study (Canadian Task Force classification II-1). SETTING: University hospital. PATIENTS: Forty-four women. INTERVENTION: Hysteroscopic myoma enucleation in toto. MEASUREMENTS AND MAIN RESULTS: With a hysteroresectoscope and Collins electrode, an elliptic incision of endometrial mucosa that covers the myoma is made at the level of its reflection on the uterine wall until the cleavage zone of the myoma is reached. Connecting bridges between myoma and surrounding muscle fibers are resected. This allows nearly complete protrusion of the myoma into the uterine cavity, facilitating complete myomectomy by slicing. The procedure was performed in 41 (93.1%) of 44 women. Of these, 38 (92.6%) had myomas between 2 and 4 cm in diameter and 3 (7.4%) had myomas exceeding 4 cm. Mean operating time was 27 minutes (range 10-45 min). CONCLUSION: This technique is efficient and allows complete resection of submucous myomas with large intramural component by favoring intracavitary protrusion of that part. PMID- 12732780 TI - Electrosurgery-induced generation of gases: comparison of in vitro rates of production using bipolar and monopolar electrodes. AB - STUDY OBJECTIVE: To compare rates of production of gases generated by VersaPoint bipolar hysteroscopic vaporizing electrodes using normal saline and monopolar vaporizing electrodes using 1.5% glycine. DESIGN: In vitro study (Canadian Task Force classification II-1). SETTING: Laboratory. MATERIAL: Bovine cardiac muscle. INTERVENTION: Fresh morbid bovine cardiac muscle was fully immersed in normal saline for the bipolar system and in 1.5% glycine for monopolar systems. Loop and bulk vaporizing electrodes were activated by radiofrequency electrosurgical units (ESUs) appropriate for each system, at standard powers and at settings higher than those in clinical use. MEASUREMENTS AND MAIN RESULTS: Rates of gas production were calculated and data analyzed by analysis of variance. For bulk vaporizing electrodes, the highest gas production rates at standard settings occurred with monopolar electrodes and the Force 2 generator at 300 W, and the Force FX at 200 W. At slightly higher than standard 200-W settings, the VersaPoint bipolar bulk vaporizing electrode was associated with lowest gas production rates in this category. Although there were statistically significant differences between loop electrodes, the magnitude remained small at similar ESU settings, and differences are thought to be clinically insignificant. CONCLUSION: Rates of gas production in this model appeared to vary most with ESU power output settings and relatively little with electrosurgical modality (bipolar or monopolar). PMID- 12732783 TI - Obstetric outcome after endoscopic transection of the uterine septum. AB - STUDY OBJECTIVE: To analyze results of endoscopic transection of uterine septum in women with primary infertility and history of recurrent abortion or late abortion. DESIGN: Observational study over 6 years (Canadian Task Force classification II-2). SETTING: University hospital. PATIENTS: Seventy women, 21 with primary infertility, 33 with recurrent abortions, and 16 with late abortion or preterm delivery. INTERVENTION: Hysteroscopic metroplasty with transection of uterine septum. MEASUREMENTS AND MAIN RESULTS: Hysteroscopic metroplasty was possibly effective for women with primary infertility, none of whom had had abortions. It was also effective in reducing the early abortion rate and in increasing term deliveries in the group with late abortion. CONCLUSION: Endoscopic transection of uterine septum may improve obstetric outcomes in women with late abortion, decrease abortion rates in those with recurrent abortions, and avoid pregnancy loss for patients with primary infertility. PMID- 12732784 TI - Colpocleisis and tension-free vaginal tape sling for severe uterine and vaginal prolapse and stress urinary incontinence under local anesthesia. AB - STUDY OBJECTIVE: To describe the technique, complications, and outcomes of vaginal repair of concomitant advanced uterine and vaginal prolapse and stress urinary incontinence using colpocleisis and tension-free vaginal tape (TVT) and pubovaginal sling under intravenous sedation and local anesthesia in elderly and/or medically compromised patients. DESIGN: Retrospective analysis (Canadian Task Force classification III). SETTING: Large tertiary care hospital with university affiliation. PATIENTS: Thirty consecutive women. INTERVENTION: Colpocleisis and TVT-pubovaginal sling. MEASUREMENTS AND MAIN RESULTS: Data were obtained by retrospective chart review of office and surgical records and follow up physical examinations. All 30 patients had the procedure completed without general anesthesia. Mean estimated blood loss was 56 ml (range 10-150 ml), mean operating time 97.3 minutes (range 65-135 min), and mean hospital stay 1.62 days (range 1-12 days). No intraoperative complications occurred, although one woman experienced a postoperative myocardial infarction. Average follow-up was 19.1 months. Three women required reoperation for minor prolapse (2 posterior repairs, 1 anteroposterior repair) and 94% were cured of stress incontinence. CONCLUSION: Preliminary data suggest that surgical correction of concomitant severe pelvic organ relaxation and stress urinary incontinence using a proved procedure (pubovaginal sling) coupled with colpocliesis can be performed rapidly and safely with local anesthesia and mild sedation, thus limiting the potential risks of general anesthesia and more invasive surgical procedures. PMID- 12732785 TI - Adhesions caused by umbilical piercing. AB - A 31-year-old woman experienced chronic pelvic pain, unresponsive to medical therapy. History, physical examination, and laboratory analyses were noncontributory except for multiple body piercings. Diagnostic laparoscopy revealed an umbilical adhesion from the small bowel to the anterior abdominal wall. The patient removed the umbilical piercing but declined surgery for adhesiolysis. PMID- 12732786 TI - Experience with 109 cases of transvaginal hydrolaparoscopy. AB - We conducted a prospective review of our experience with 109 transvaginal hydrolaparoscopies (THLs) performed in 97 women. The THL was considered complete in 101 procedures (93%) and adequate for management in 105 (96%). Two complications occurred (1.8%), one failed entry and one perforation of a retroflexed uterus. Diagnoses for 67 infertile patients were normal pelvis in 34 (51%), endometriosis in 14 (21%), adhesions in 6 (9%), and tubal obstruction in 10 (15%); 3 THLs (4%) were considered incomplete. Of 17 women with dysmenorrhea, a normal pelvis was found in 8 (47%) and endometriosis in 9 (53%). In 11 patients with pelvic pain endometriosis was found in 4 (36%), normal pelvis in 3 (27%), and adhesions in 3 (27%); THL was incomplete in 1 (9%). Six infertile patients (9%) had operative laparoscopy and 10 (15%) operative THL; 6 (9%) were counseled to seek in vitro fertilization. Pregnancy occurred in 16 patients (24%). Analog pain scores (0 = no pain, 10 = worst pain) were tracked in 39 consecutive patients. Pain during trocar insertion averaged 4.2 +/- 0.5, 2.2 +/- 0.2 at midprocedure, and 1.1 +/- 0.1 at the end of THL. We believe that THL should be considered instead of hysterosalpingogram and laparoscopy in selected patients. PMID- 12732787 TI - Fundamentals of peritoneal access. PMID- 12732788 TI - Is laparoscopic high McCall colpopexy effective in treating uterine prolapse with uterine preservation? PMID- 12732789 TI - Does the editorial board live up to September 10, 2001, expectations? PMID- 12732791 TI - C-reactive protein: a guideline for its application. PMID- 12732792 TI - Gender differences on the risk evaluation of acute coronary syndromes: the CARDIO2000 study. AB - Coronary heart disease (CHD) is more common in men than women. Gender differences in CHD risk may be explained by a different impact that coronary risk factors may have for men and women, in the development of CHD. Thus, the authors aimed to analyze the extent to which cardiovascular risk factors can explain the gender difference in CHD risk, at population level. During 2000-2001, 848 hospitalized patients with a first event of acute coronary syndrome and 1078 controls, paired by gender, age, and region with no evidence of overt CHD, were randomly selected from all Greek regions. Data revealed that women experiencing their first acute coronary syndrome were significantly older than men (65.3+/-8 vs. 59.7+/-10 years old; p<0.01), and that acute coronary syndrome occurred more frequently in men than women (frequency ratio 4:1, men:women). When adjusting for age, multivariate analysis revealed that both family history of premature CHD and hypercholesterolemia were associated with higher coronary risk in men than women (odds ratio [OR]=5.11 vs. 3.14; p<0.05 for family history and OR=3.77 vs. 2.19; p<0.05 for hypercholesterolemia). The presence of hypertension however, had a significantly greater effect in women than men (OR=4.86 vs. 1.66; p<0.01). Also, higher education level and the adoption of a Mediterranean diet had a more protective effect in women than men (OR=0.53 vs. 0.87; p<0.001; and OR=0.80 vs. 0.96; p<0.05, respectively). There was also evidence of a greater association between depression and higher coronary risk in women than men (OR=1.93 vs. 1.58; p<0.07). The impact of other factors (i.e., smoking, diabetes, body mass index, physical activity, alcohol consumption, and financial status), on the coronary risk difference between genders was similar for men and women. In conclusion, our findings suggest that the contribution of certain coronary risk factors to the risk for CHD is different for men and women. PMID- 12732793 TI - Screening for diabetes in general practice. AB - The prevalence of type 2 diabetes is rising rapidly worldwide. Evidence suggests that between one third and one half of cases are undiagnosed and patients may have preclinical disease for as long as 12 years. At diagnosis, 50% of patients have microvascular complications (retinopathy, neuropathy, or nephropathy) and patients have twice the risk of macrovascular disease compared to the background population. Screening for type 2 diabetes would allow earlier recognition of cases, with the potential to intervene earlier in the disease course, but whether this would result in improved long-term outcomes is unknown. The debate continues about who should be considered for screening, how we should screen, and whether we should screen for diabetes at all. The authors review the evidence, particularly in light of the recent position statement on diabetes screening published by the American Diabetes Association. If we do start screening for diabetes, one of the major challenges ahead is to ensure resources are in place to allow optimization of treatment for the increasing number of patients. This is important both in those found to have diabetes and in those with lesser degrees of glucose intolerance who are at high risk of developing diabetes and are at increased risk of macrovascular disease. PMID- 12732794 TI - Treatment of atherosclerosis in the new millennium: is there a role for vitamin E? AB - Oxidative stress appears to be of fundamental relevance to diseases as diverse as atherosclerosis, cancer, and Alzheimer's disease. Observational data in humans have suggested that antioxidant vitamin intake is associated with reduced cardiovascular disease. Animal studies are largely consistent with the concept that dietary supplementation with antioxidant vitamins reduces the progression of atherosclerosis. However, recent prospective, controlled clinical trials of vitamin E, including the Cardiovascular Disease, Hypertension and Hyperlipidemia, Adult-Onset Diabetes, Obesity, and Stroke (CHAOS) study, the Heart Outcomes Prevention Evaluation (HOPE) trial, Gruppo Italiano per lo Studio della Sopravvivvenza nell'Infarto Miocardico (GISSI)-Prevenzione trial, the Secondary Prevention with Antioxidants of Cardiovascular Disease in End Stage Renal Disease (SPACE) trial, and the Heart Protection Study (HPS) present a confused picture. The various possibilities that have been advanced to explain this discrepancy are discussed in this review. A striking feature of these and other trials of antioxidants is the absence of a biochemical basis for patient inclusion or, indeed, dose selection. Patients with high levels of oxidant stress or depletion of natural antioxidant defense systems may be the most likely to benefit from antioxidant therapy. If this is the case, then reliable, quantitative indices of in vivo oxidant stress such as urinary isoprostane levels should be considered as an inclusion criterion for patient selection. Future trials of antioxidant therapy in cardiovascular disease should then be targeted toward such patients with high levels of oxidant stress or patients with depletion of natural antioxidant defense systems. Furthermore, the dose of antioxidant should be chosen based on a surrogate readout that is a reliable, reproducible, and easily obtainable in vivo measure of oxidant stress. In the interim, although the safety of vitamin E up to doses of 800 IU/day has been determined, the conflicting nature of the results published to date encourages us to avoid making premature recommendations with respect to vitamin E supplementation in the prevention and treatment of cardiovascular disease. PMID- 12732795 TI - Alcohol use and prognosis in patients with coronary heart disease. AB - Substantial evidence has shown that moderate drinkers have lower rates of coronary heart disease (CHD) than abstainers, but the effects of alcohol consumption among patients with established CHD are less clear. Alcohol intake has important effects on risk factors for reinfarction, including higher levels of high-density lipoprotein cholesterol and triglycerides, lower levels of fibrinogen and other prothrombotic factors, lower fibrinolytic potential, and antiplatelet activity. Studies of patients at risk for CHD, such as those with diabetes, hypertension, and hyperlipidemia, have shown that the association of moderate drinking and CHD is at least as strong as it is in the general population. Most recently, studies have found that survivors of acute myocardial infarction who drink moderately have a risk of death approximately 20%-30% lower than do abstainers or rare drinkers. Nonetheless, the risks and benefits of alcohol use remain complex, even among patients with CHD, and no simple recommendation regarding alcohol consumption can be made for all patients. PMID- 12732796 TI - Implication of earlier carotid atherosclerosis for stroke and its subtypes. AB - In addition to advanced stenosis, earlier stages of carotid atherosclerosis are associated with the risk for stroke. Based on a recent report, this review will focus on the associations between carotid atherosclerosis and stroke subtypes. The authors report increased severity of carotid atherosclerosis in atherothrombotic and lacunar infarction patients, compared to nonstroke patients. However, the severity was similar between nonstroke patients and those with cardioembolic infarction and cerebral hemorrhage. By receiver operating characteristic curve analyses, although evaluation of carotid atherosclerosis appears to help for risk assessment of atherothrombotic and lacunar infarction, the potential for benefit may be greater for atherothrombotic. Large prospective studies are still necessary to establish the link between earlier carotid atherosclerosis and the future risk for specific stroke subtypes. PMID- 12732797 TI - Impact of age on definition of standards for ideal weight. AB - There has been an ongoing debate regarding the impact of age while establishing standards for ideal weight. Based on current federal guidelines released by the National Heart, Lung, and Blood Institute, ideal weight is defined as a body mass index between 18.7-24.9 for all adults regardless of age. However, studies failed to show a positive association between overweight--as defined by the current guidelines--and all-cause and cardiovascular mortality in individuals 65 years and older. Data suggest that optimum body mass index tended to be higher for older adults compared with the young and middle-aged population, and interestingly, being heavier than recommended by the current guidelines resulted in the lowest mortality rates. Therefore, allocation of time and resources for weight reduction interventions among the mild- to moderately-overweight elderly is not advisable. To establish standards for ideal weight, future guidelines should consider age-specific recommendations that reflect the results of recent studies. PMID- 12732798 TI - LIFE and ARBITER. PMID- 12732799 TI - Symptom reflections of women with cardiac disease and advanced practice nurses: a descriptive study. AB - The purpose of this descriptive study was to explore the experiences of women with cardiac disease in regard to cardiac risk factors, symptoms, and symptom interpretation. Interviews with a convenience sample of 21 African American and Caucasian female patients, aged 40-81 years with cardiac diagnoses, were taped and transcribed. Three advanced practice nurses with experience in cardiovascular nursing were interviewed individually for background. Analyses of symptoms indicated that women experienced a range of prodromal symptoms other than chest pain prior to hospitalization such as: edema, shortness of breath, lightheadedness, dry cough, and exhaustion. Atypical symptoms delayed recognition of a cardiac problem for some women. The lack of symptom consistency underscores the need for thorough assessment and screening of women at risk for cardiac problems. Advanced practice nurses are aware of current cardiac disease research in women but they must continue to be vigilant for atypical symptoms. PMID- 12732800 TI - Preliminary reliability and validity of a family caregiver conflict scale for stroke. AB - Recovery from stroke is a challenging process for stroke survivors and their families. Assessing family conflict may be useful in guiding interventions to reduce caregiver distress. The purpose of this study was to assess preliminary reliability and validity data for a 15-item family caregiver conflict scale [FCCS] for stroke. Construct validity was assessed using subscales from the family assessment device: communication, problem solving, general family functioning, and a perceived criticism scale. Data were pooled from three studies of caregivers of stroke survivors (total N=93). Internal consistency for the FCCS was adequate (>0.70). Higher family conflict was related to more ineffective family communication, general family functioning, and higher perceived criticism (p > or =0.05). Nonspouse caregivers reported more family conflict than spouse caregivers. The FCCS appears to have adequate internal consistency and construct validity and may provide information about family conflicts around stroke recovery and direction for family-focused interventions. PMID- 12732801 TI - Pulmonary assessment: what you need to know. AB - The purpose of this review is to describe a step-by-step assessment of the pulmonary system. Beginning with the sequence of the history and physical examination related to pulmonary assessment, the author describes the most common chief complaints and potential causes of these symptoms. The inspection, palpation, percussion, and auscultation steps of the physical examination are also reviewed. A detailed assessment of the pulmonary system will aid in the identification of patient problems and assist in the selection of appropriate goals and interventions for the patient. PMID- 12732802 TI - Transitioning adolescents with congenital heart disease into adult health care. AB - The effort and commitment dedicated to the treatment of children with congenital heart disease over the past 50 years has been astounding. Therefore, it is imperative to ensure these young patients who have survived the varying challenges of their conditions a smooth transition into a new world of adult health care. The key to the long-term management of an increasing number of children surviving congenital heart disease is the effective coordination of care from pediatric to adult health care. Strategies for facilitating the transition of the adolescent with congenital heart disease into adult health care practices are presented in this paper. PMID- 12732803 TI - T wave alternans: a marker of myocardial instability. AB - T wave alternans is an electrocardiographic marker of myocardial electrical instability considered predictive for the development of lethal ventricular rhythms. Direct observation of this phenomenon on the electrocardiogram is rare. Subtle, nonvisible alternation of the T wave may be detected at the microvolt level using advanced signal processing techniques. This technology has enabled investigators and clinicians to identify and risk-stratify certain groups at high risk for sudden cardiac death. The purpose of this review is to discuss the concept of T wave alternans in relation to all cardiac alternans, provide an overview of its proposed mechanisms, and address the clinical utility of this electrocardiographic marker of myocardial electrical instability. PMID- 12732804 TI - Dilemmas in drug therapy. PMID- 12732805 TI - Can a 12-lead ECG be used to assess for hyperkalemia? PMID- 12732806 TI - Principles of population-based genetic screening and their importance. PMID- 12732807 TI - Anticoagulation at elderly age: the challenge to do better. PMID- 12732809 TI - Clinical diagnosis of aortic stenosis in the elderly is difficult. PMID- 12732811 TI - Management of the elderly aortic stenosis patient with low gradient and low ejection fraction. AB - The incidence of aortic stenosis increases with age and thus it occurs frequently in elderly patients. Once severe obstruction has developed, death occurs within 3 years unless the aortic valve is replaced. The results of aortic valve surgery, even in octogenarians, are usually excellent in the absence of comorbidity. The exception to this rule is for the aortic stenosis patient who has low ejection fraction, a low cardiac output and a transvalvular gradient of <30 mm Hg. Such patients have far advanced left ventricular dysfunction and increased operative mortality. However, even these patients may benefit from surgery if they have truly severe aortic stenosis. Because valve area is unreliable at low cardiac outputs, output should be increased pharmacologically in such patients and the valve area recalculated. If the transvalvular gradient increases with output, severe aortic stenosis is present and valve replacement may be beneficial. However, if output increases but gradient does not, only mild stenosis is present and surgery is unlikely to prolong life. PMID- 12732812 TI - Pathophysiology of valvular aortic stenosis in the elderly. AB - Aortic stenosis in the elderly is related to calcification of either a bicuspid valve or a morphologically normal tricuspid valve. There is increasing evidence that factors relating to atherosclerosis are involved in valvular calcification and that it is an actively regulated process rather than a degenerative one. With severe aortic stenosis left ventricular hypertrophy occurs, decreasing wall stress and supporting the left ventricular ejection fraction. However, with pathologic hypertrophy there is a dropout of myocardial cells, subendocardial ischemia, and fibrosis. Eventually, symptoms of angina, non-Q wave myocardial infarction, exertional syncope, and heart failure occur. Once symptoms begin, the prognosis is poor, with sudden death occurring in about one third of patients who die. In the elderly, symptoms can be recognized very late in the course of the disease since they can be attributed to other problems and since the elderly patient may have reduced physical activity to a minimum. The more comorbidities that exist, the greater the risk of valve replacement. Symptomatic patients with severe aortic stenosis even over age 80 can be operated upon with a relatively low mortality and morbidity. In patients over age 80, prolongation of life for any meaningful length of time is not as important as relief of symptoms and improvement in the quality of life. Thus, it is unlikely that any truly asymptomatic patient over age 80, even with severe aortic stenosis, should be sent to surgery. PMID- 12732813 TI - Calcific aortic stenosis in the elderly: a brief overview. AB - Aortic stenosis is the most common valvular lesion in patients above the age of 65. The etiology, presentation, and management of aortic stenosis differs in the elderly compared to younger patients in many ways. Many of the classic physical findings are absent in the elderly, making the diagnosis of critical aortic stenosis more difficult. Due to coexisting morbidity in many elderly patients, there is often a reluctance to recommend aortic valve replacement despite the dismal prognosis of medical therapy. In the following review, the authors discuss the pathophysiology, presentation, and management of aortic stenosis in the elderly. PMID- 12732814 TI - Infective endocarditis in the elderly: diagnosis and management. AB - Infective endocarditis is seen with increasing frequency in older patients. This increase is due to the general aging of the population, improved survival of patients with congenital and valvular heart disease, and the increasing use of catheters and other prosthetic devices with resulting higher incidence of nosocomial endocarditis. In older patients, infective endocarditis frequently develops in the absence of underlying structural heart disease; atheromatous deposits and mitral annular calcification are two important risk factors in this population. Infective organisms in older patients are frequently enterococci and other gastrointestinal tract bacteria. A marked febrile response is uncommon whereas central nervous system symptoms are more common in older patients. Transesophageal echocardiography can be performed safely and is a major diagnostic tool with sensitivity of more than 90% in detecting vegetations as small as 2-5 mm. Appropriately drawn blood cultures provide bacteriologic diagnosis in 80%-99% of patients. Prolonged antibiotic therapy may be required in many instances depending on the infective organism. Early surgical therapy is advisable for patients who develop heart failure as a result of severe acute aortic valvular regurgitation. Valve replacement surgery can be performed with acceptable mortality and morbidity even in very elderly patients. PMID- 12732815 TI - Is there any indication for aortic valvuloplasty in the elderly? AB - Elderly patients with severe symptomatic calcific aortic stenosis do poorly with medical management. The optimal treatment for this group of patients is surgical valve replacement. Balloon valvuloplasty may be useful as a bridge to aortic valve replacement in hemodynamically unstable patients, in patients undergoing emergent noncardiac surgery, and in patients with severe comorbidities who are too ill to undergo cardiac surgery. Balloon valvuloplasty often results in symptomatic improvement; however, the postvalvuloplasty valve area is usually <1.0 cm2, the major periprocedural complication rate is roughly 5%, and the 6 month restenosis rate is quite high. There is no evidence that balloon valvuloplasty alters the natural history of aortic stenosis, although no randomized study has been performed. PMID- 12732816 TI - Prophylactic aortic valve replacement in older patients for mild aortic stenosis during coronary bypass surgery. AB - Coronary bypass surgery (CBS) is performed in many older patients who frequently also have mild calcific aortic stenosis. It is important that a correct assessment of the severity of aortic stenosis is done by calculating the aortic valve area. Mild aortic stenosis is aortic valve area >1.5 cm(2), >0.9 cm(2)/m(2); severe aortic stenosis is aortic valve area < or =1.0 cm(2), < or =0.6 cm(2)/m(2). Patients who have severe aortic stenosis should have aortic valve replacement (AVR) at the time of CBS. Patients with mild aortic stenosis should not have AVR simultaneously with CBS because: 1) patients having AVR+CBS have a higher operative and 10-year mortality; 2) prosthetic heart valves are associated with a complication rate of 2%-6% per year; and 3) only about 12% of patients with mild aortic stenosis will have developed severe aortic stenosis in 10 years. Performing AVR for mild aortic stenosis at the time of CBS will probably result in 91 unnecessary AVRs and 29 excess deaths in 10 years. PMID- 12732818 TI - Ethical issue: The patient undergoes surgery that she knows will not benefit her, hoping that she will die in the process while under anesthesia. PMID- 12732817 TI - Diagnosis of aortic stenosis in the elderly: role of echocardiography. AB - Doppler echocardiography provides accurate hemodynamic information for the diagnosis, assessment and follow-up of patients with aortic stenosis, making diagnostic cardiac catheterization redundant in most cases. Considering the rapid growth of the aging population and the high incidence of aortic stenosis, as the most common valvular heart disease in the age group, the authors have described the utility of the Doppler technique with a brief discussion of the clinical evaluation of these patients. Close observation of patients with asymptomatic aortic stenosis has been emphasized. Decision-making about the appropriate timing for aortic valve replacement is of immense clinical significance, which may be facilitated and achieved by the periodic follow-up and the use of this noninvasive method. Following a comprehensive echo-Doppler evaluation, the patient would only need coronary arteriography prior to surgery. The utility of hemodynamic and anatomic information by echocardiography in this type of surgery is emphasized. PMID- 12732819 TI - Three-dimensional transesophageal echocardiographic assessment of aortic valve pathology. PMID- 12732821 TI - [Infectious diarrhoeas] AB - The infective diarrhoea show a high morbility in infants of all countries and a high mortality in infants and children of developing countries. In developed countries there is a high frequency of "traveler's diarrhoea". The different pathogenesis of infectious diarrhoeas (intestinal invasion, intestinal adherence, citonecrotic or citotonic toxins) influences the clinical presentation (watery diarrhoea or diarrhoea with blood and mucos, presence or assence of fever). The key factor in the management of acute watery diarrhoea is the restoration of water and electrolyte balance; but frequently an antimicrobial treatment is needed. PMID- 12732820 TI - MADIT II: lessons to learn. PMID- 12732823 TI - [Neurobrucellosis: clinical and therapeutic features] AB - Brucellosis is a common zoonosis endemic in many parts of the world. Neurologic involvement is a rare but serious complication and it occurs in 2 - 5% of cases. Neurologic signs can appear during the active phase of disease or later. Six cases of neurobrucellosis are described: meningoencephalitis in 2 cases, meningitis with brain abscess in 1 case, encephalomielitis in 3 cases. Diagnosis was made on the basis of history of the exposure to brucella, of serum and cerebrospinal fluid positive serology, of quantitative change in cerebrospinal fluid, on EEG, CT and MNR and favorable response to specific treatment (rifampin, rolitetracycline tirmethoprim-sulfamethoxazole). Neurobrucellosis should be suspected in case of neurological deficit in brucellosis endemic areas. Treatment of neurobrucellosis remains controversial: we recommend a regimen with rifampin 600 mg x 2 i.v. + tirmethoprim-sulfamethoxazole 2 ff x 2 i.v. for a period of 6-8 weeks. PMID- 12732822 TI - [Treatment of Community Acquired Pneumonia (CAP) in adults with amoxicillin/clavulanic acid switch therapy: a multicenter italian study] AB - Patients with community acquired pneumonia (CAP) previously treated with other oral antibiotics (Cephalosporins 35,3%, Macrolides 26,9%, Tetracyclines 14,1%, Quinolones 12,8%, other 10,9%) for at least 72 hours at home were considered for this national multicentre study. 43 hospitalized patients (17 males and 26 females) ranging between the ages of 19 and 79 were treated with Amoxicillin/Clavulanic acid (AMC) 1,2 g e.v., T.l.D. for at least 3 days. I.V. Treatments were switched to AMC p.o. after apyrexial status. Average treatment duration was 10,8 + 3.6. Sputum/B.A.L. samples were obtained from all patients submitted for microbiological exams in order to determine microbiological aetiology of CAP in patients who have failed on previous antibiotics. At the end of treatment, 30 patients (96,8%) were considered cured, while treatment failed in 1 patient (3,2%). 12 patients were considered not evaluable because serology was positive for M.pneumoniae (5 pts); for C.pneumoniae (3 pts.); 1 patients was positive for Mycobacterium tuberculosis; 1 patient was positive for HIV and Mycobacterium spp.; 2 patients were non-evaluable for lung or bronchial tumor. Not reported adverse events. PMID- 12732824 TI - [Assuring quality of AIDS care: organizational and functional aspects] AB - The implementation of a method to assess the quality of care provided to HIV infected subjects and AIDS patients is outlined. Two sets of factors have been considered: one pertaining the structural aspects of the health care setting ("D. Cotugno" Hospital, the largest infectious diseases hospital in Southern Italy); the other concerning the operational and qualitative aspects of care, including staff and patients' satisfaction. Structural factor have been assessed by analyzing data provided by hospital administration; operational aspects have been investigated by means of medical record review and the administration of a semi structured interview to health care workers. Results helped to prioritize evidence-based interventions aimed to improve the quality of AIDS care. PMID- 12732825 TI - [Multiple osteo-articular involvement due to methicillin-resistant Staphylococcus aureus septicaemia: clinical and therapeutic evaluation] AB - Here we report a rare case of septic spondilodiskitis by methicillin-resistant Staphylococcus aureus, complicated by the atypical involvement of two articular sites such as manubrio-clavicular joints and right wrist. The source of the septic process was identified in hand's eczematous lesions and paronychia. A first therapeutical attempt performed by combining teicoplanin with netilmicin or rifampicin was useless. A new course with vancomycin instead of teicoplanin favoured the prompt remission of symptoms. Following 10 weeks of continuous treatment, we observed the complete disappearance of all radiological signs of vertebral damage. Though rarely, polyarthritis may complicate a Staphylococcus aureus bacteraemia. An adequate chemio-antibiotic course may lead to definitive recovery and avoid surgery. PMID- 12732826 TI - [Rhodococcus equi pneumonia in AIDS: a case report] AB - We describe the first case, in Campania, of Rhodococcus equi pneumonia in an HIV positive subject. The clinical symptoms resolved after antibiotic therapy. The pulmonary lesion at 10 months of radiological follow up results just partially reduced in its diameters. PMID- 12732827 TI - Type I necrotising fasciitis of heel in a diabetic patient. AB - Necrotising fasciitis is a potentially fatal clinical disease caused by infection with various bacteria. Diabetes mellitus, one of the predisposing factors for necrotising fasciitis is also an important factor in the progression of the illness by increasing its morbidity and mortality rate. In this case report, a 54 year old diabetic woman with type-I necrotising fasciitis of the heel was treated with extensive debridments, broad spectrum antimicrobial therapy and intensive regulation of blood glucose levels. The resulting new positive data pertaining to the morbidity and mortality of this severe disease suggest that the old management of this disease should be re-examined. PMID- 12732828 TI - The struggle against the variolous poison (Part 1). AB - Not available PMID- 12732829 TI - Giving thanks for nurses in difficult times. PMID- 12732830 TI - What are older people prepared to do to avoid falling? A qualitative study in London. AB - Falls prevention strategies form a significant part of recent government policy relating to older people. In this article we report a qualitative study to examine the extent to which older people are willing to adopt any of the following strategies in order to avoid falling; balance and lower limb strengthening exercises, home safety advice, and 'taking care'. A sample of community-dwelling inpatients who were more or less afraid of falling were interviewed. Some older people blamed their own unsafe behaviour rather than hazards in their home for falls, so taking care (moving slowly, holding on, being vigilant to and avoiding danger) was a widespread strategy. The benefits of exercise in falls prevention were poorly understood and there was little enthusiasm for exercising especially if it was perceived to be inappropriate in old age or painful. Awareness of home hazards and taking measures against them was common but home visits tended to be regarded as intrusive. Inadequate negotiation about adaptations was widespread and led to resentment and unwillingness to cooperate. Building on the widespread 'taking care' strategy may help educated older people in safe behaviour. Health professionals should encourage older people to exercise and when making home safety visits should confer with their clients, taking their preferences about adaptations into account. PMID- 12732831 TI - Health visiting at the crossroads: which way to the 21st century? AB - This article presents a response from a group of lecturers at Florence Nightingale School of Nursing and Midwifery, King's College London and one service user to the perception of a crisis of confidence within health visiting. Changes in registration procedures and shifts in policy have led health visiting to question its place within primary care and the model of preparation for health visiting practice. The group argues that this debate is based on a misunderstanding of the changing health care environment and that health visiting can find a place within the primary care team. Key to this is the continued adherence to a basic nursing training. The authors suggest an incremental model of health visitor training to provide the fullest scope to implement the new health agenda. PMID- 12732832 TI - Indwelling catheter-associated urinary tract infections. AB - Catheter associated urinary tract infection accounts for a large proportion of all hospital-acquired infections, and there has been little change in the incidence of such infections since the introduction of closed sterile urinary drainage systems in the 1960s. The aim of this paper is to review the literature on the subject of catheter associated urinary tract infections, with reference to the use and possible improvement of closed sterile catheter drainage systems. PMID- 12732833 TI - Preventing blockage of long-term indwelling catheters in adults: are citric acid solutions effective? AB - Community health practitioners are frequently faced with the problem of indwelling catheters failing to drain. This mini-review examines the evidence regarding the effectiveness of citric acid-based bladder washout solutions (solution G, solution R), compared with saline for preventing blockage of long term catheters in older adults. Although there is some evidence on the effect of citric acid solutions in laboratory conditions, the search yielded no evidence measuring catheter patency in humans. Only one randomized controlled trial addressing the use of citric acid solutions with human subjects was found. This provided some confirmation that solution G had immediate effect but gave no evidence of long-term benefit. No difference was apparent on examination of catheter tips. As the evidence is weak, practitioners should consider the use of citric acid maintenance solution as non-routine and only following formal evaluation in a one-off trial on appropriate patients. PMID- 12732834 TI - Developing a culture-specific tool to assess postnatal depression in the Indian community. AB - Postnatal depression is a serious and debilitating condition which affects, at a conservative estimate, 10% of postnatal mothers. It can be difficult to identify due to the reluctance of some women to acknowledge their feelings. The Edinburgh Postnatal Depression Scale was developed to facilitate the elicitation of negative mood, however, it does not translate adequately into other languages or cultures. To address this problem the dosha assessment tool for postnatal depression was developed. This tool is aimed at women who are more familiar with the concepts of Ayurvedic medicine, one of the key medical systems of the Indian subcontinent. Although not yet evaluated, the tool goes some way towards the development of a culturally specific screening tool for this condition. PMID- 12732835 TI - Older people as health service consumers 4: disempowered or disinterested? AB - Giving patients a greater say in the NHS is prominent in all national service frameworks (NSFs) and is evident in the expert patient initiative (Department of Health (DH), 2001a). One group of users who may have difficulty in making their voice heard are the elderly. The national service framework for older people (DH, 2001b) promotes user involvement in an attempt to achieve fair and equitable access to services for this group of patients who, statistically, make the highest use of the NHS and could be classified as consumers. This article, the last in a series on this topic, discusses whether a sample of older people in a village community have a consumerist ethos in regard to health care, and whether they are prepared to act in this way. Suggestions are made for better facilitating older people's involvement in and use of services. PMID- 12732836 TI - Does research reach practice? PMID- 12732837 TI - Public health: a new nursing role for community practitioners. AB - This article discusses the six key activities identified by the Royal College of Nursing (1994) as essential ingredients for public health nursing. Examples of public health practice and community nurse involvement are given to illustrate each activity. Public health skills such as health profiling, building healthy alliances and policy development are outlined. Current barriers to community nurses participating in these activities are identified and models of public health practice are reviewed. The potential for a radical transformation of community nursing practice is discussed in the context of an expanding public health role of community nurses in Scotland. Options for future practice are presented and discussed. PMID- 12732838 TI - Drug therapy in the elderly: improving quality and access. PMID- 12732839 TI - "Inactive" excipients such as Cremophor can affect in vivo drug disposition. PMID- 12732841 TI - Effect of simultaneous versus staggered dosing on pharmacokinetic interactions of protease inhibitors. AB - OBJECTIVE: The aim of this study was to determine whether pharmacokinetic interactions between the protease inhibitors saquinavir soft gel, nelfinavir, and ritonavir are affected by the timing of administration. STUDY DESIGN: We used an open-label, 6-period, incomplete Latin square crossover study in 18 human immunodeficiency virus-negative subjects. Each received single oral doses of 2 of the 3 protease inhibitors during each of 6 periods. Single doses were given either simultaneously or separated by 4 hours. The order of the periods was balanced, and periods were separated by 2 days. We measured protease inhibitor concentrations over a 24-hour period by HPLC and estimated pharmacokinetic parameters by noncompartmental methods. RESULTS: Median saquinavir area under the curve (AUC) increased by 62-fold when ritonavir was coadministered, by 50-fold when ritonavir was given 4 hours earlier, and by 16-fold when saquinavir preceded ritonavir by 4 hours. Saquinavir AUC increased by 7-fold when nelfinavir was coadministered. Nelfinavir AUC increased by 2.5-fold with coadministration of ritonavir and by 1.8- and 2.1-fold when ritonavir was administered before nelfinavir and after nelfinavir, respectively. Ritonavir AUCs were unaffected by coadministration of the other drugs. The effect of ritonavir on the kinetics of saquinavir persisted for at least 48 hours after a single dose of ritonavir, suggesting the possibility of metabolic intermediates that form inhibitory complexes. CONCLUSION: Except for saquinavir followed by ritonavir, there is little difference in protease inhibitor exposure for simultaneous or staggered doses. The persistent effect of ritonavir suggests the possibility that lower doses and longer dosing intervals might be effective when ritonavir is used to boost concentrations of other protease inhibitors. PMID- 12732840 TI - Pharmacokinetic and pharmaceutic interaction between digoxin and Cremophor RH40. AB - BACKGROUND: The pharmacokinetics of digoxin is modulated by the efflux pump P glycoprotein. Cremophor EL (BASF Aktiengesellschaft, Ludwigshafen, Germany) (polyoxyl 35 castor oil), a castor oil derivative used to improve the solubilization of drugs and vitamins, has been shown to inhibit this membrane transporter in vitro and in vivo. So far, no study in humans has evaluated the effect of Cremophor RH40 (BASF Aktiengesellschaft) (polyoxyl 40 hydrogenated castor oil) on P-glycoprotein. METHODS: A randomized, double-blind, placebo controlled crossover study in 12 healthy individuals was performed with a single oral dose of 0.5 mg digoxin in a hard gelatin capsule in combination with multiple doses of oral Cremophor RH40 (600 mg 3 times daily) or placebo. A digitized electrocardiogram with 12 standard leads was recorded to assess the pharmacodynamics of digoxin. RESULTS: Cremophor RH40 delayed and enhanced the absorption of digoxin in the first 5 hours after dosing. During Cremophor RH40 administration, digoxin lag time was significantly prolonged compared with placebo (0.53 +/- 0.25 hour versus 0.36 +/- 0.19 hour, P =.04). The peak concentration of digoxin increased by 22%, from 2.21 +/- 0.94 ng/mL to 2.69 +/- 1.28 ng/mL (P =.06). Similarly, the area under the plasma concentration-time curve from 0 to 5 hours significantly increased by 22% (5.23 +/- 1.63 h. ng/mL versus 4.30 +/- 1.12 h. ng/mL, P =.03). Digoxin did not cause a clinically significant change in the dynamic parameters during both periods. CONCLUSION: This study demonstrates a pharmacokinetic and pharmaceutic interaction between the emulgent Cremophor RH40 and digoxin, caused by P-glycoprotein inhibition and prolongation of the dissolution time of digoxin tablets by Cremophor RH40, respectively. Our in vivo study in humans supports the validity of in vitro observations on P-glycoprotein. PMID- 12732842 TI - Cisplatin tumor concentrations after intra-arterial cisplatin infusion or embolization in patients with oral cancer. AB - BACKGROUND: One neoadjuvant course of intra-arterial high-dose cisplatin (cis diamminedichloroplatinum [CDDP]) tumor perfusion combined with intravenous sodium thiosulfate (STS) (cisplatin neutralizer) infusion is part of a multimodality concept for treatment of oral cancer. Recently, crystalline cisplatin embolization has been described as a novel treatment variant with increased tumor response rates. METHODS: We have compared tumor and plasma concentrations of cisplatin and STS by means of microdialysis in 10 patients with oral cancer treated with intra-arterial cisplatin perfusion (150 mg/m(2) in 500 mL of 0.9% sodium chloride) and 6 patients with oral cancer treated with crystalline cisplatin embolization (150 mg/m(2) in 45-60 mL of 0.9% sodium chloride), respectively. The microdialysis catheter was placed into the tumor, and the intra arterial catheter into the tumor-feeding artery. Cisplatin was rapidly administered through the intra-arterial catheter and STS (9 g/m(2)) was infused intravenously to reduce the systemic toxicity of cisplatin. STS infusion was started 10 seconds after the cisplatin infusion was started. RESULTS: After embolization, cisplatin tumor maximum concentration (C(max)) and tumor area under the concentration-time curves (AUCs) were about 5 times higher than those achieved after intra-arterial perfusion (C(max), 180.3 +/- 62.3 micromol/L versus 37.6 +/- 8.9 micromol/L), whereas the opposite was true for plasma concentrations (C(max), 0.9 +/- 0.2 micromol/L versus 4.7 +/- 0.6 micromol/L). STS plasma levels were about 3 times higher than its tumor concentrations (C(max) tumor, 1685 +/- 151 micromol/L; C(max) plasma, 5051 +/- 381 micromol/L). After the standard intra arterial perfusion, the average STS/CDDP AUC ratios for tumor and plasma were 211 +/- 75 and 984 +/- 139, respectively. After cisplatin embolization, the respective ratios were 48.5 +/- 29.5 and 42966 +/- 26728. CONCLUSION: Molar STS/CDDP ratios of greater than 500 are required outside the tumor to neutralize cisplatin, whereas tumor ratios should be lower than 100 to avoid a loss of tumor cell killing. The first goal is achieved with both treatment modalities and the second only with cisplatin embolization, suggesting that crystalline cisplatin embolization is superior to intra-arterial cisplatin perfusion in terms of tumor cisplatin concentrations. Whether this translates into higher tumor response rates needs to be investigated further. PMID- 12732843 TI - Characterization of hepatic cytochrome p4503A activity in patients with end-stage renal disease. AB - BACKGROUND: The cytochrome p450 (CYP) oxidative enzyme system, located primarily in the liver and small intestine, is responsible for metabolism and detoxification of numerous endogenous and exogenous substances. The most abundant CYP enzyme, CYP3A, is known to be involved in the metabolism of more than 200 commonly used medications. In experimental models of renal failure, both hepatic function and CYP enzyme content are reduced; however, direct evidence in humans is lacking. Evaluation of drug metabolism in patients with end-stage renal disease is important because these patients use a large number of medications and are at risk of adverse reactions and drug-drug interactions. METHODS: We measured hepatic CYP3A activity at baseline and after rifampin (INN, rifampicin) enzyme induction in 12 patients with end-stage renal disease and 12 healthy, age-matched controls. Hepatic CYP3A phenotype was characterized with the erythromycin breath test, and enzyme induction capacity was evaluated with a short course of rifampin (600 mg/d for 6 days). RESULTS: The end-stage renal disease group had 28% lower baseline erythromycin breath test values than controls (P <.05); however, enzyme induction capacity after rifampin administration was similar between groups (P =.70). CONCLUSION: The findings suggested that one mechanism by which patients with end-stage renal disease are at increased risk of drug toxicity is reduced activity of the CYP3A enzyme pathway. PMID- 12732844 TI - A study to survey susceptible genetic factors responsible for troglitazone associated hepatotoxicity in Japanese patients with type 2 diabetes mellitus. AB - BACKGROUND AND OBJECTIVE: Troglitazone is a 2,4-thiazolidinedione antidiabetic agent with insulin-sensitizing activities. This agent had been used efficiently in a large number of patients but was withdrawn from the market in March 2000 because of its association with idiosyncratic hepatotoxicity. To address the susceptible genetic factors responsible for the hepatotoxicity associated with this agent, we performed a genetic polymorphic analysis by a target gene approach in troglitazone-treated Japanese patients with type 2 diabetes mellitus. METHODS: One hundred ten patients treated with troglitazone were recruited into this study. The case patients (n = 25) were recruited through medical professionals who had previously reported abnormal increases in the levels of ALT or AST among their patients. The control patients (n = 85) were recruited through physicians prescribing troglitazone. For statistical accuracy, efforts were made to maximize the size of the case group. Genotype analysis was performed in 68 polymorphic sites of 51 candidate genes related to drug metabolism, apoptosis, roduction and elimination of reactive oxygen species, and signal transduction pathways of peroxisome proliferator-activated receptor gamma 2 and insulin. RESULTS: The strong correlation with transaminase elevations was observed in the combined glutathione-S-transferase GSTT1-GSTM1 null genotype (odds ratio, 3.692; 95% confidence interval, 1.354-10.066; P =.008). CONCLUSIONS: The double null mutation of GSTT1 and GSTM1 might influence troglitazone-associated abnormal increases of liver enzyme levels. PMID- 12732846 TI - Polymorphisms in the CYP1A2 gene and theophylline metabolism in patients with asthma. AB - BACKGROUND: Cytochrome p450 (CYP) 1A2 gene polymorphisms are thought to be involved in theophylline metabolism. OBJECTIVE: We analyzed the effect of genetic polymorphisms in the 5'-flanking region to the first intron of the CYP1A2 gene on theophylline metabolism in 75 Japanese patients with asthma and 159 healthy Japanese volunteers. METHODS: Genetic polymorphisms were detected at 4 sites of the CYP1A2 gene, -2964(G/A) and -1569(T/del) in the 5'-flanking region and 155(T/G) and 731(A/C) in the first intron. RESULTS: There were no significant differences in the distribution of gene polymorphisms between the asthmatic and control groups. Among asthmatic patients, theophylline clearance was significantly lower in patients with the polymorphism at site -2964(G/A) whose genotype was G/A (0.029 +/- 0.001 L x h(-1) x kg(-1)) or A/A (0.029 +/- 0.002 L x h(-1) x kg(-1)) than in those whose genotype was G/G (0.034 +/- 0.001 L x h(-1) x kg(-1)) (P <.01 and P <.05, respectively). High theophylline clearance levels significantly correlated with age in the G/G subgroup of site -2964(G/A) (P <.05, r = -0.35) but not in the G/A or A/A subgroup. CONCLUSION: Given its potential side effects, theophylline may need to be used with care in patients with the A allele at site -2964(G/A) in the CYP1A2 gene, because theophylline metabolism levels are lower in such patients, particularly in young asthmatic individuals. PMID- 12732845 TI - Coordinated intrahepatic and extrahepatic regulation of cytochrome p4502D6 in healthy subjects and in patients after liver transplantation. AB - Cytochrome p450 (CYP) 2D6 activity exhibits wide intersubject variation even among individuals with similar genotypes in whom the active enzyme is expressed. There is, therefore, a need to understand the mechanisms involved in determining its activity. The relationship of messenger ribonucleic acid (mRNA) expression to CYP2D6 activity has been evaluated in hepatic and extrahepatic tissues to test the hypothesis of coordinated regulation. In human liver microsomes, there was a greater than 25-fold variation in both bufuralol hydroxylation and concentration of mRNA for CYP2D6, with a significant association between variables (n = 20; Spearman correlation coefficient [r(s)] = 0.85, P <.001). In normal subjects, there was a similar extent of interindividual variation in in vivo activity of CYP2D6, measured as the debrisoquin (INN, debrisoquine) recovery ratio, and in mRNA for CYP2D6 in peripheral blood mononuclear cells, with a significant association between variables (n = 78; r(s) = 0.56 [95% confidence interval, 0.35 to 0.73], P <.001), whereas no association was found between mRNA for CYP2D6 and CYP2E1 activity. Recipients of liver transplants, at a time of stable liver function, had a similar relationship between debrisoquin recovery ratio and concentration of mRNA for CYP2D6 in peripheral blood mononuclear cells (n = 27; r(s) = 0.74 [95% confidence interval, -0.16 to 0.44], P <.001). Three recipients, who had CYP2D6*4/*4 genotypes, remained phenotypically poor metabolizers for CYP2D6 after liver transplantation. Collectively, these results imply that transcriptional regulation of mRNA for CYP is a major determinant of in vivo activity and that regulation of intrahepatic and extrahepatic enzymes is coordinated, possibly through a mechanism that is predominantly extrahepatic. PMID- 12732847 TI - Response to "The efficacy of simvastatin is not influenced by CYP2D6 polymorphism" by Geisel et al. PMID- 12732848 TI - Cytochrome P4502C9 activity in end-stage renal disease. PMID- 12732849 TI - The need to consider the cost factor of drugs in clinical trials. PMID- 12732851 TI - Beat-to-beat analysis of the relation between RT and RR intervals in newborns. AB - OBJECTIVE: To evaluate the dynamic RT (QRS apex-end of T wave) rate dependence in newborns. STUDY DESIGN: A Digital Holter ECG was acquired on day 15 in nine full term and eight preterm infants. Ten-minute periods were recorded during wakefulness and sleep. The accuracy of fit with RT-RR pairs was individually assessed by 14 regression formulas (r coefficient, Akaike score, residual analysis). The medians of RT and Bazett's RT correction were calculated for each 10 milliseconds of RR. RESULTS: The mean RR and RT were 429+/-51 and 263+/-18 milliseconds. None of the prediction formulas were sufficiently accurate to describe RT over the whole range of RR (r<0.56). The Bazett correction produced differences of more than 50 milliseconds at different RR. Prematurity, sleep state and heart rate variability did not influence RT-RR relation. CONCLUSION: None of the parametric formulas were found to be accurate in describing RT rate dependence in newborns. PMID- 12732852 TI - Defining the true pathogenesis and pathophysiology of neonatal encephalopathy and cerebral palsy. PMID- 12732850 TI - Humanin peptide suppresses apoptosis by interfering with Bax activation. AB - Bax (Bcl2-associated X protein) is an apoptosis-inducing protein that participates in cell death during normal development and in various diseases. Bax resides in an inactive state in the cytosol of many cells. In response to death stimuli, Bax protein undergoes conformational changes that expose membrane targeting domains, resulting in its translocation to mitochondrial membranes, where Bax inserts and causes release of cytochrome c and other apoptogenic proteins. It is unknown what controls conversion of Bax from the inactive to active conformation. Here we show that Bax interacts with humanin (HN), an anti apoptotic peptide of 24 amino acids encoded in mammalian genomes. HN prevents the translocation of Bax from cytosol to mitochondria. Conversely, reducing HN expression by small interfering RNAs sensitizes cells to Bax and increases Bax translocation to membranes. HN peptides also block Bax association with isolated mitochondria, and suppress cytochrome c release in vitro. Notably, the mitochondrial genome contains an identical open reading frame, and the mitochondrial version of HN can also bind and suppress Bax. We speculate therefore that HN arose from mitochondria and transferred to the nuclear genome, providing a mechanism for protecting these organelles from Bax. PMID- 12732854 TI - Hyperglycemia and retinopathy of prematurity in very low birth weight infants. AB - OBJECTIVE: Retinopathy of prematurity (ROP) remains a leading cause of morbidity in the very low-birth-weight (VLBW) infant. This study investigates a possible association between serum/blood glucose and the development of ROP. METHODS: A retrospective case-control study of all infants born between 1992 and 1997 at the Johns Hopkins Hospital with birth weights less than 1000 g who developed Stage 3 or 4 ROP was conducted. Controls either had Stage 1 ROP or no eye disease and were matched 2:1 with ROP patients for gestational age, birth weight and year of birth. Odds ratios (ORs) of ROP were calculated for multiple exposures over the first month after birth, including oxygen concentration (FiO(2)), blood glucose levels, vitamin E, mean airway pressure and mean blood pressure. RESULTS: In a simple logistic regression analysis, we found an increased ROP risk for: (1) each 10 mg/dl increase of mean glucose (OR 1.96; 95% CI 1.13 to 3.42), (2) each 1% increase of mean FiO(2) (OR 1.06; 95% CI 1.004 to 1.13), (3) history of dopamine infusion (OR 5.4; 95% CI 1.16 to 25.2) and (4) intraventricular hemorrhage Grade 3 or 4 (OR 7.3; 95% CI 1.53 to 34.7). Using a multiple regression model, we found an increased ROP risk for each 10 mg/dl increase of mean glucose (OR 2.7; 95% CI 1.003 to 7.27). Each IU/kg/day of vitamin E supplementation reduced ROP risk (OR 0.37; 95% CI 0.16-0.86). CONCLUSION: In this study, we could demonstrate that glucose levels in the first month of life are associated with the development of ROP. Further studies have to determine if this association is causal or if hyperglycemia is just an expression of severity of illness. PMID- 12732853 TI - Proton magnetic resonance spectroscopy improves outcome prediction in perinatal CNS insults. AB - OBJECTIVE: Prediction of neurologic outcome is difficult in neonates with acute nervous system injury. Previous studies using proton magnetic resonance spectroscopy ((1)H-MRS) have been used to predict short-term neurologic outcome in neonates with a variety of neurologic insults. We were interested in determining the effectiveness of combining clinical evaluation and spectroscopy obtained at the time of injury in predicting neurologic outcome at 24 months. STUDY DESIGN: We studied 33 neonates with acute central nervous system injury, 5.8+/-3.7 days of injury, owing to hypoxic-ischemic encephalopathy. Neonates were assessed using clinical variables (initial arterial pH, initial blood glucose, Sarnat score, electroencephalography) and spectroscopy (NAA/Cho, NAA/Cre, Cho/Cre, and lactate). Neonates were divided into two outcome groups: good/moderate and poor. Differences between the groups were assessed using chi(2) and t-test analyses. We analyzed the best predictors of outcome using discriminant analysis and calculated sensitivity, specificity, positive, and negative predictive values for each variable independently and in combination. RESULTS: There were significant differences between the good/moderate and poor outcome for the Sarnat score, EEG, lactate, and NAA/Cho. Spectroscopy combined with clinical variables improved sensitivity, but not specificity for predicting outcome. The presence of lactate had the best individual predictive value. Combination of the clinical with the MRS variables had the highest predictive value. CONCLUSION: Proton magnetic resonance spectroscopy done early after injury improves the ability to predict neurologic outcome at 24 months of age. PMID- 12732855 TI - Early bubble CPAP and outcomes in ELBW preterm infants. AB - OBJECTIVE: To test whether the introduction of early bubble continuous positive airway pressure (CPAP) results in improved respiratory outcomes in extremely low birth-weight infants. STUDY DESIGN: Outcomes of all infants between 401 and 1000 g born in a level 3 neonatal intensive care units (NICU) between July 2000 and October 2001 (period 2) were compared using historical controls (period 1). Early bubble (CPAP) was prospectively introduced in the NICU during period 1. Univariate and adjusted comparisons were made across time periods. RESULTS: Delivery room intubations, days on mechanical ventilation and use of postnatal steroids decreased (p<0.001) in period 2, while mean days on CPAP, number of babies on CPAP at 24 hours (p<0.001) and mean weight at 36 weeks corrected gestation also increased (p<0.05) after introduction of early bubble CPAP. CONCLUSIONS: Early bubble CPAP reduced delivery room intubations, days on mechanical ventilation, postnatal steroid use and was associated with increased postnatal weight gain with no increased complications. PMID- 12732856 TI - Tolerance of a sterile isotonic electrolyte solution containing select recombinant growth factors in neonates recovering from necrotizing enterocolitis. AB - OBJECTIVE: To assess the tolerance of a sterile isotonic electrolyte solution containing select recombinant growth factors enterally administered in neonates who were NPO because of necrotizing enterocolitis (NEC). STUDY DESIGN: A phase I trial was accomplished among 30 neonates. Patients received 5, 10, or 20 mL enterally of the study solution/kg/day divided into every 3-hour dosing, for 3 days prior to when feedings of milk were to resume. The occurrence of emesis, gastric residuals, diarrhea, bloody stools, abdominal distention, skin rashes and death were sought. RESULTS: Gestational ages ranged from 25.2 to 41.1 weeks. A total of 16 neonates had Stage IA NEC, six Stage IB, and eight Stage IIA. The solution was well tolerated in all 30; none developed diarrhea, guaiac positive or bloody stools, or abdominal distention. Administration of the solution was not prematurely discontinued in any infant. Two neonates died secondary to late-onset sepsis remote from the study period. CONCLUSIONS: Enteral administration of a sterile isotonic electrolyte solution containing select recombinant growth factors was well tolerated by neonates with NEC. PMID- 12732857 TI - Usefulness of the minute ventilation test in predicting successful extubation in newborn infants: a randomized controlled trial. AB - OBJECTIVE: We performed a prospective, randomized clinical trial to compare the usefulness of the minute ventilation test (MVT) with clinical judgement in predicting readiness for extubation in preterm newborns with respiratory distress syndrome requiring surfactant therapy and mechanical ventilation. STUDY DESIGN: A total of 42 preterm infants with respiratory distress syndrome were randomized when they reached preselected ventilator settings. The primary outcome measure was the time from study entry to extubation, provided the infant remained extubated for at least 24 hours. RESULTS: Infants evaluated by the MVT were extubated in a significantly shorter period of time (mean of 8 hours) than those evaluated clinically (mean of 36 hours). The extubation failure rate was similar in the two groups. CONCLUSION: The MVT is an easily performed objective measure that can be used to predict readiness for extubation in preterm infants. In this study, it significantly shortened the time for extubation and was not associated with a higher rate of reintubation. PMID- 12732858 TI - Does elevated peak bilirubin protect from retinopathy of prematurity in very low birthweight infants. AB - OBJECTIVE: Bilirubin is a potent in vitro antioxidant. Despite repeated study, its in vivo significance has yet to be defined. Bilirubin is universally elevated in very low birthweight (VLBW) infants. Retinopathy of prematurity (ROP) is a disease thought to be associated with exposure to oxygen free radicals in VLBW infants. The objective of this study was to determine whether there was an association between peak bilirubin levels and ROP. METHODS: The risk for ROP, stages III and IV was measured as a function of increasing peak bilirubin levels in VLBW infants admitted to the neonatal ICU. A similar analysis was performed on a subgroup of VLBW infants with prolonged (> or =28 days) oxygen requirement. The relation between peak bilirubin level and the duration of oxygen requirement was tested by logistic regression analysis. All analyses were conducted after controlling for birthweight and presence of intraventricular hemorrhage (IVH). RESULTS: There was an increased risk for ROP, stages III and IV (OR 1.187; 95% CI 1.013 to 1.390; p=0.034) with elevated peak serum bilirubin levels in the entire population. Duration of oxygen requirement was not related to peak bilirubin (p>0.1). In the subgroup of infants with prolonged oxygen requirement (> or =28 days), there was no association between peak serum bilirubin levels and ROP III and IV (p>0.1); however, there was an association with further increased duration of oxygen requirement (p=0.034). CONCLUSION: Elevated peak bilirubin does not protect from and may be a risk for ROP in VLBW infants. PMID- 12732859 TI - Closed suctioning of intubated neonates maintains better physiologic stability: a randomized trial. AB - OBJECTIVE: To evaluate the physiological variance in a closed (CS) vs an open suction (OS) protocol in intubated infants. STUDY DESIGN: Infants were stratified into three weight groups in a randomized crossover trial. Heart rate, respiratory rate, blood pressure, oxygen saturation, transcutaneous oxygen and carbon dioxide, and end-tidal carbon dioxide were recorded prior to suctioning, during suctioning, and recovery to baseline. Following the procedures, recovery time to baseline parameters was measured. Data were analyzed using repeated measures ANOVA. RESULTS: Overall, there was significantly less deviation from baseline physiological parameters with CS. Infants <1000 g had clinically significant decreases in heart rate with the OS method (-18% OS vs -6% CS; p=0.016). Recovery time in the OS group was twice that of the CS cohort (4 vs 2 minutes; p<0.001). CONCLUSION: CS maintains better physiologic stability in intubated infants. PMID- 12732860 TI - A prospective controlled study of neonatal morbidities in infants born at 36 weeks or more gestation to Women with diet-controlled gestational diabetes (GDM class Al). AB - OBJECTIVE: Infants of gestational diabetes mellitus (GDM)-A1 women are unlikely to experience the marked excursion in maternal glucose levels that may characterize insulin-requiring GDM (class-A2) or insulin-dependent diabetes (IDDM). However, infants born to GDM-A1 women are traditionally managed like infants born to GDM-A2 or IDDM women. AIMS: To examine monitoring protocols for infants of GDM-A1 women, and to examine the efficacy of early and frequent feedings to prevent and to treat hypoglycemia. METHODS: A total of 92 of 101 infants born to GDM-A1 women (diabetic group) and 68 of 83 infants born to nondiabetic women (control group) at > or=36 weeks of gestation were prospectively monitored for the development of hypoglycemia and other morbidities. Blood glucose screening was performed in the diabetic group every 30 60 minutes three times, starting soon after birth and then at 3-hour intervals for 24 hours. Liberal feedings were started shortly after birth and provided every 3 hours for at least 24 hours. All women with GDM-A1 had an HbA1c measured before delivery. RESULTS: Both the diabetic and control groups had similar demographics, including LGA incidence. Blood glucose readings before feedings were low (<40 mg/dl) in 24 of 92 infants (26.1%) from the diabetic group and in 20 of 68 control infants (29%). After the start of oral feedings, all but four diabetic and three control infants had subsequent glucose readings > or =40 mg/dl. No infant had symptoms of hypoglycemia and none from the diabetic group had birth trauma, hypoxic-ischemic encephalopathy, polycythemia, hypocalcemia, or hypomagnesemia. Hypoglycemic episodes in the infants from the diabetic group could be managed with oral feedings alone. Birth weight, gestational age, sex, Apgar scores, and maternal HbA1c levels could not predict low glucose readings on initial screening in infants from the diabetic group. CONCLUSION: The incidence of hypoglycemia in infants born to GDM-A1 women at > or =36 weeks of gestation is similar to control infants born to nondiabetic women. Low blood glucose levels during the first few hours of life can be prevented or treated with early and frequent oral feeding. PMID- 12732861 TI - Clinical dilemma in triplet pregnancy: when is it appropriate to intervene for a jeopardized fetus? AB - OBJECTIVE: To determine gestational age-specific risks of intervening to "rescue" a compromised fetus in triplet pregnancies. STUDY DESIGN: We analyzed retrospectively triplet pregnancies managed at New England Medical Center (July 1992-May 2000; n=97 pregnancies). For each week in gestation, we compared the chance of at least one of three infants developing complications of prematurity in Scenario A (delivery at that gestation to rescue the jeopardized fetus) with the chance of at least one of two infants from Scenario B (allowing the jeopardized fetus to die in utero to prolong pregnancy) developing that complication later in gestation. RESULTS: We observed a decreased risk of at least one infant developing a specific complication in Scenario B than in Scenario A for all complications studied. CONCLUSIONS: Comparison of triplet outcomes with the two surviving older newborns identifies important changes in risk between 25 and 32 weeks. These data enable physicians and parents to weigh acceptable risks with benefits. PMID- 12732862 TI - Expectant management of midtrimester premature rupture of membranes: a plea for limits. AB - OBJECTIVE: Our aim was to assess neonatal and maternal complications of the expectant management of pregnancies with preterm premature rupture of membranes (P-PROM) prior to 24 weeks of gestation and to delineate a patient consult strategy. STUDY DESIGN: We included all consecutive cases of early midtrimester P PROM (16-24 weeks gestation). Information coded in our perinatal database was analyzed. Descriptive statistics, Student's t-test and Mann-Whitney test, and a logistic regression model were built accordingly. RESULTS: A total of 28 women presented with P-PROM at 16-24 weeks (mean 22.7+/-1.0 weeks). Two patients declined conservative management and one was lost to follow-up (10.7%). In all, 25 (89.2%) were followed until the onset of labor or development of chorioamnionitis. Overall, 8/25 (32%) Of the neonates survived. Pulmonary hypoplasia accounted for three deaths (3/25, 12%). Of 10 pregnancies with P-PROM before 22 weeks gestation, two (20%) neonates survived. The amount of amniotic fluid and gestational age at the time of diagnosis were crucial independent factors determining overall survival. Pulmonary hypoplasia (12%) and skeletal deformities (0%) were infrequent. The 21-day mean maternal antenatal hospital stay was further complicated by a high cesarean rate delivery (33.7%) and by postpartum infectious morbidity (32%). CONCLUSION: In cases of early midtrimester P-PROM (<24 weeks) expectantly managed, neonatal survival is positively associated with the amount of amniotic fluid present and with the gestational age at the time of diagnosis. The mothers are at increased risk of prolonged antenatal hospitalization, cesarean delivery, preterm birth, and postpartum infection. In very early midtrimester P-PROM (<22 weeks), the maternal complication rate outweighs the poor neonatal outcome and expectant management should be reconsidered. PMID- 12732863 TI - Reflections on errors in neonatology III. The "experienced" years, 1970 to 2000. PMID- 12732865 TI - Neonatal acute renal failure secondary to maternal exposure to telmisartan, angiotensin II receptor antagonist. AB - Fetal and neonatal toxic effects of angiotensin II receptor antagonists have been described in animals and humans. Five cases of fetal or neonatal deaths have been reported following maternal use of sartans for hypertension. We report a case of neonatal transient renal failure following telmisartan therapy during pregnancy. This class of antihypertensive drugs should be avoided during pregnancy and breastfeeding. PMID- 12732864 TI - Hyperbaric oxygen therapy for a neurologically devastated child: whose decision is it? AB - A recent case highlights one of the on-going and unresolved controversies in pediatric ethics: who makes treatment decisions for children. Children, by definition, do not have the maturity to make medical choices. Those decisions must be made for them. The issue remains by whom and on what standard those choices should be made. PMID- 12732866 TI - Aspergillosis in a 24-week newborn: a case report. AB - Aspergillosis is an uncommon neonatal infection, diagnosed with an increasing frequency over the last two decades. We report a premature neonate who developed aspergillosis while receiving amphotericin B and fluconazole for candidiasis. Despite early recognition and diagnosis, the infant died. We review the clinical appearance of Aspergillus species, the distinctions between primary cutaneous aspergillosis and invasive aspergillosis, and advances in diagnosis and treatment. PMID- 12732868 TI - Targeting 'horror autotoxicus' by gene therapy: results, challenges and the future. PMID- 12732869 TI - Celiac disease: a model autoimmune disease with gene therapy applications. AB - Gene therapy (GT) is still at the 'experimental' stage and some recent setbacks have cooled the potential use of this therapeutic tool even in life-threatening conditions. However, this therapeutic approach has a potential, which is not limited to disease for which we have not other option. There are increasing evidence that GT will be soon used in diseases that are not life threatening. One group of diseases that can benefit from GT is the autoimmune one. Several experimental animal models have indicated the efficacy (proof of principle) of GT. In the present review, we have addressed the possibility that even extremely benign autoimmune-like diseases such as Celiac Disease (CD) might one day profit from this type of therapy. We further point that in conditions such as CD, where the trigger is well known and the pathogenic cascade is relatively well defined, a situation not common in autoimmunity, we can even have a better situation where to explore and use GT to control disease initiation and progression. Once the risks that are still intrinsic to GT will have been reduced the therapeutic options we outline in the present review might not appear too far from reality. PMID- 12732870 TI - Gene therapy in autoimmune, demyelinating disease of the central nervous system. AB - Multiple sclerosis (MS) is an immune-mediated disease of the central nervous system (CNS), where suspected autoimmune attack causes nerve demyelination and progressive neurodegeneration and should benefit from both anti-inflammatory and neuroprotective strategies. Although neuroprotection strategies are relatively unexplored in MS, systemic delivery of anti-inflammatory agents to people with MS has so far been relatively disappointing. This is most probably because of the limited capacity of these molecules to enter the target tissue, because of exclusion by the blood-brain barrier. The complex natural history of MS also means that any therapeutic agents will have to be administered long-term. Gene therapy offers the possibility of site-directed, long-term expression, and is currently being preclinically investigated in experimental autoimmune encephalomyelitis (EAE), an animal model of MS. While some immune effects may be targeted in the periphery using DNA vaccination, strategies both viral and nonviral are being developed to target agents into the CNS either via direct delivery or using the trafficking properties of cell-carrier systems. Targeting of leucocyte activation, cytokines and nerve growth factors have shown some promising benefit in animal EAE systems, the challenge will be their application in clinical use. PMID- 12732871 TI - Gene transfer approaches for the treatment of inflammatory bowel disease. AB - The pathogenesis of Crohn's disease and ulcerative colitis, the two major forms of inflammatory bowel disease, involves a complex interplay between certain genetic, environmental and immunological factors. Considerable research progress in the last decade defined key inflammatory pathways in the inflamed gut and identified new potential therapeutic targets. Since the current medical treatment with corticosteroids and anti-inflammatory drugs is often associated with undesired side effects and cannot completely cure IBD, these current advances in our understanding of intestinal pathology may now allow the development of new biologic treatment strategies including gene therapy. In this review, we will give a brief overview of potential gene therapy target molecules related to chronic intestinal inflammation. Furthermore, we summarize the results of recent preclinical studies for intestinal gene transfer and discuss future perspectives. PMID- 12732873 TI - Gene- and cell-based therapeutics for type I diabetes mellitus. AB - Type 1 diabetes mellitus, an autoimmune disorder is an attractive candidate for gene and cell-based therapy. From the use of gene-engineered immune cells to induce hyporesponsiveness to autoantigens to islet and beta cell surrogate transplants expressing immunoregulatory genes to provide a local pocket of immune privilege, these strategies have demonstrated proof of concept to the point where translational studies can be initiated. Nonetheless, along with the proof of concept, a number of important issues have been raised by the choice of vector and expression system as well as the point of intervention; prophylactic or therapeutic. An assessment of the current state of the science and potential leads to the conclusion that some strategies are ready for safety trials while others require varying degrees of technical and conceptual refinement. PMID- 12732872 TI - Immunopathology and the gene therapy of lupus. AB - Lupus is a chronic autoimmune inflammatory disease with complex clinical manifestations. In humans, lupus, also known as systemic lupus erythematosus (SLE), affects between 40 and 250 individuals, mostly females, in each 100 000 of the population. There are also a number of murine models of lupus widely used in studies of the genetics, immunopathology, and treatment of lupus. Human patients and murine models of lupus manifest a wide range of immunological abnormalities. The most pervasive of these are: (1) the ability to produce pathogenic autoantibodies; (2) lack of T- and B-lymphocyte regulation; and (3) defective clearance of autoantigens and immune complexes. This article briefly reviews immunological abnormalities and disease mechanisms characteristic of lupus autoimmunity and highlight recent studies on the use of gene therapy to target these abnormalities. PMID- 12732874 TI - Gene therapy in nonhuman primate models of human autoimmune disease. AB - Before autoimmune diseases in humans can be treated with gene therapy, the safety and efficacy of the used vectors must be tested in valid experimental models. Monkeys, such as the rhesus macaque or the common marmoset, provide such models. This publication reviews the state of the art in monkey models for rheumatoid arthritis and multiple sclerosis and the (few) gene therapy experiments that have been performed in these models. PMID- 12732875 TI - Gene therapy for arthritis. AB - Rheumatoid arthritis is an autoimmune disease with intra-articular inflammation and synovial hyperplasia that results in progressive degradation of cartilage and bone, in severe cases it causes systemic complications. Recently, biological agents that suppress the activities of proinflammatory cytokines have shown efficacy as antiarthritic drugs, but require frequent administration. Thus, gene transfer approaches are being developed as an alternative approach for targeted, more efficient and sustained delivery of inhibitors of inflammatory cytokines as well as other therapeutic agents. Indeed, the efficacy of gene transfer for the treatment of arthritis has been demonstrated in mouse, rat, rabbit, and horse models of disease whereas the feasibility of the approach has been demonstrated in Phase I clinical trials. In this review, the current status of both preclinical and clinical arthritis gene therapy is presented. In addition, the advantages and disadvantages of different types of vectors, target cells and therapeutic genes being developed for the treatment of arthritis are summarized. Finally, the future directions of the rapidly developed field of arthritis gene therapy are outlined. PMID- 12732876 TI - Vectors for the treatment of autoimmune disease. AB - Gene therapy has been applied in a variety of experimental models of autoimmunity with some success. In this article, we outline recent developments in gene therapy vectors, discuss advantages and disadvantages of each, and highlight their recent applications in autoimmune models. We also consider progress in vector targeting and components for regulating transgene expression, which will both improve gene therapy safety and empower gene therapy to fullfil its potential as a therapeutic modality. In conclusion, we consider candidate vectors that satisfy requirements for application in the principal therapeutic strategies in which gene therapy will be applied to autoimmune conditions. PMID- 12732877 TI - Engineering mesenchymal stem cells for immunotherapy. AB - Allogeneic hematopoietic stem cell transplantation, after sublethal irradiation of recipient animals, is capable of inducing donor-specific tolerance facilitating subsequent organ transplantation. This approach could reintroduce tolerance in autoimmune diseases and it has been applied to treat autoimmune diseases with, however, a great susceptibility of recurrence. Mesenchymal stem cells (MSCs) present within the bone marrow could be critical to the immunosuppressive effect of the treatment. This tolerance induction may be useful in allogeneic transplantations, where low incidence of graft-versus-host disease was observed when the hematopoietic graft was coinjected with MSCs. In this paper, we discuss the use of MSCs in different therapeutic strategies either as immunosuppressive agents or genetically engineered to express molecules acting against the autoimmune process. PMID- 12732878 TI - 'GVHD': graft-versus-host disease or graft-versus-Hodgkin's disease? An old acronym with new meaning. AB - The majority of patients with relapsed or refractory Hodgkin's lymphoma (HL) will not be cured with standard therapy. Relapse rates remain high even after autologous stem cell transplantation (SCT), particularly for patients with high risk disease. Allogeneic SCT offers several potential advantages for patients with HL. It is feasible when autologous stem cells are not available and stem cell grafts will be tumor free. Perhaps a more important advantage is the potential to generate a graft-versus-Hodgkin's lymphoma (GVHL) effect. Unfortunately, although allogeneic SCT may cure some HL patients, treatment related mortality has been unusually high, and superior survival, when compared to autologous SCT, has not been demonstrated. Nonmyeloablative conditioning and allogeneic SCT may induce a direct GVHL reaction with less conditioning regimen related toxicity and ultimately may have the potential to improve cure rates and survival for advanced HL patients. PMID- 12732879 TI - Autologous stem cell transplantation for acute myeloid leukemia. AB - Autologous bone marrow transplant (ABMT) and stem cell transplantation (ASCT) are important treatment modalities for acute myeloid leukemia (AML). The role of ASCT in first remission patients remains controversial. Phase II and phase III studies demonstrate that patients with favorable-risk cytogenetics benefit from ASCT, with reduction in relapse and improvement in leukemia-free survival (LFS). Patients with poor-risk cytogenetics do not appear to benefit significantly from ASCT and should preferentially be treated with allogeneic transplant. The role of ASCT for patients with intermediate risk disease is uncertain. It appears that ASCT in first remission will improve disease-free survival compared to standard chemotherapy. Sufficient patients who relapse after chemotherapy treatment can be salvaged with ASCT in second remission such that the beneficial effect on overall survival is blunted. ASCT produces equivalent results to ABMT but with reduced morbidity. The collection of stem cells during recovery from intensive dose consolidation therapy appears to be an attractive strategy that can increase the percentage of patients who are able to receive their intended transplant. Consolidation therapy prior to stem cell collection and transplant has been shown to decrease the relapse rate and improve outcomes, but the optimal nature of this consolidation therapy is unknown. For patients with AML in second remission, ABMT/ASCT offers a substantial salvage rate, and is particularly effective for patients with acute promyelocytic leukemia. PMID- 12732880 TI - G-CSF alone vs cyclophosphamide plus G-CSF in PBPC mobilization of patients with lymphoma: results depend on degree of previous pretreatment. AB - We performed a randomized study to compare 'G-CSF alone' (administered at dose of 10 mcg/kg/day) and 'cyclophosphamide plus G-CSF' (cyclophosphamide at dose of 4 g/m(2) and G-CSF at dose of 10 microg/kg/day), as PBPC mobilization schedules in 52 patients with NHL or HD. Randomization was stratified according to the amount of previous chemotherapy (< or =2 and >2 lines of previous chemotherapy). Mean CD34+ cell peak in P.B., mean 'Total CD34+ cells' harvested and percentage of patients successfully mobilized, in the group mobilized with 'G-CSF alone' vs the group mobilized with 'cyclophosphamide plus G-CSF', were: 35.3 x 10(6) vs 45.8 x 10(6)/l (P=0.3), 5.4 x 10(6) vs 6.8 x 10(6)/kg (P>0.9) and 50 vs 61% (P=0.4). No differences were observed in the stratum of less pretreated patients. However, in the stratum of patients who had previously received more than two lines of chemotherapy, CD34+cell peak (P=0.05) and percentage of successful mobilization (P=0.01) were higher when 'cyclophosphamide plus G-CSF' was used. Using logistic regression, both age and mobilization with 'G-CSF alone' were significantly associated with a low CD34+ cell peak in P.B. However, in the stratum of less pretreated patients, only age was significantly associated with this risk. PMID- 12732881 TI - High-dose thiotepa, melphalan and carboplatin (TMCb) followed by autologous stem cell transplantation in patients with advanced breast cancer: a retrospective evaluation. AB - This study was conducted to evaluate the efficacy of high-dose thiotepa, melphalan and carboplatin (TMCb) regimen in 27 patients undergoing autologous stem cell transplantation (ASCT) for metastatic breast cancer. A total of 27 patients with stage IV breast cancer underwent ASCT following thiotepa (500 mg/m(2)), melphalan (100 mg/m(2)) and carboplatin (1200-1350 mg/m(2)). Of 27 patients, 17 had refractory relapse, eight had responding relapse, and two had no evidence of disease (NED) at the time of transplant. In all, 11 patients had only bone disease, nine had bone plus visceral disease, three had only visceral disease, and two had locoregional recurrent disease. The median time from diagnosis to transplant was 1081 days (range 180-2341). Staging for evaluation of response was performed 4-6 months after transplantation. Five patients were not evaluable (NE) for response because of NED at transplant (n=2) or early death due to transplant-related complications (n=3) (two of viral pneumonia and one of regimen-related toxicity) occurring at a median of 4 days (range 11-46) post transplant. One of the two patients who was NED at the time of transplant is still NED on day 760 post-transplant. Seven of 15 refractory (47%) and 5/7 (71%) responsive patients with evaluable disease achieved a complete response of all measurable disease or all soft-tissue disease with at least improvement in bone lesions. Of 27 patients (37%),(10) are alive and progression-free, a median of 582 days (range 410-1380) after treatment, 6/17 (35%) with refractory disease and 4/10 (40%) with responsive disease. The probability of progression-free survival (PFS) for all patients was 0.50. The probabilities of PFS at 2 years for patients with refractory (n=17) and responsive (n=10) disease were 0.42 and 0.60, respectively. PFS at 2 years for the 14 patients who were NED or achieved CR/PR(*) following-HDC was 0.67. PFS at 2 years for patients who did not achieve CR/PR(*) following-DHC was 0.33. These preliminary data suggest that high-dose TMCb followed by autologous stem cell transplantation is an effective regimen for patients with advanced breast cancer and may be comparable to some previously used regimens. PMID- 12732882 TI - High-hyperdiploidy in Philadelphia positive adult acute lymphoblastic leukaemia: case-series and review of literature. AB - Patients with Philadelphia positive (Ph(+)) adult acute lymphoblastic leukaemia (ALL) have a poor prognosis. Stem cell transplantation (SCT) is increasingly being recognised as the treatment of choice in eligible patients with Ph(+)ALL, but disease-relapse remains a problem in a proportion of patients prior to and after SCT. Genetic abnormalities in addition to the Ph chromosome may influence the biology and clinical course of ALL, but there are not many studies on the potential genetic heterogeneity of adult Ph(+)ALL and clinical outcomes. Here, we report on five patients with ALL who were double Ph(+) and also had a high hyperdiploid karyotype (Ph(+)/hyperdiploid) at diagnosis. In contrast to the presence of the Ph(+) chromosome, high-hyperdiploidy (>50 chromosomes) as the sole karyotypic abnormality in ALL is associated with a favourable clinical outcome. In our series, four patients with a Ph(+)/hyperdiploid karyotype achieved a cytogenetic remission after induction chemotherapy and proceeded to stem cell transplantation (SCT). The fifth had five subclones including Ph(+)/hyperdiploid, as well as those that were only Ph(+); the latter emerged as the dominant clone after induction therapy and the patient died of disease relapse. Of the patients who underwent SCT, only one relapsed, but achieved a durable remission with donor lymphocyte infusions. Thus, it is conceivable that the presence of high-hyperdiploidy as an additional karyotypic abnormality may confer a better prognosis to Ph(+)ALL, presumably by altering the kinetics of Ph(+) neoplastic cells. We have discussed these results in the context of recent studies on the significance of high-hyperdiploidy in Ph(+) adult ALL. PMID- 12732883 TI - High rate of remission and low rate of disease recurrence in patients with multiple myeloma allografted with PBSC from their HLA-identical sibling donors. AB - A total of 30 multiple myeloma patients (M=23, F=7; age 31-55 years, median 48) were allografted with peripheral blood stem cells (PBSC) from HLA-identical siblings. Time to transplantation was 3-107 months (median 8). Prior chemotherapy lines varied from 1 to 6 (median 1). Four patients were in complete remission (CR), 11 in partial remission (PR), 13 were considered to be nonresponders, and two had progressive disease. Most were conditioned with busulfan-melphalan. PBSC were collected by apheresis after G-CSF or sequential GM-CSF and G-CSF. The patients were grafted with 4.4-24.1 x 10(6)/kg CD34+ (median 7.9) and 0.9-7.9 x 10(8)/kg CD3+ cells (median 2.3). GVHD prophylaxis was methotrexate-cyclosporine. Engraftment was complete and rapid. Grades II-IV acute GVHD (aGVHD) developed in 16 (53%), but was grade III-IV only in five (17%); chronic GVHD (cGVHD) developed in 17 out of the 24 evaluable patients (71%). A total of 18 patients (71%) attained CR after transplantation. TRM was 30% overall, 16% at 100 days. There was only one relapse. Overall survival and event-free survival at 73 months were 60% and 67%, respectively. PCR negativity for IgH-gene rearrangement occurred in all persistently CR patients studied. PBSC allograft can induce long remissions, because of profound suppression of the neoplastic clone that is probably linked to the antitumor effect of cGVHD. PMID- 12732884 TI - Safety and feasibility of CHOP/rituximab induction treatment followed by high dose chemo/radiotherapy and autologous PBSC-transplantation in patients with previously untreated mantle cell or indolent B-cell-non-Hodgkin's lymphoma. AB - Patients with no prior chemotherapy and with advanced and progressive follicular lymphoma (FCL) or mantle cell lymphoma (MCL) were enrolled into a treatment protocol combining CHOP/rituximab-CHOP therapy with subsequent consolidation high dose therapy (HDT) to evaluate the safety and feasibility of this treatment. Overall, 15 patients were enrolled and 13 patients completed the entire treatment protocol without major toxicities or increased infectious complications. One patient withdrew consent after achieving complete remission (CR) prior to HDT. One patient was taken off study with signs of disease progression after induction treatment. All patients showed stable engraftment after HDT. Response rates appear to be favorable, indicating an additional effect of rituximab and HDT. Overall, 12 of 13 patients achieved CR/CRu and one patient partial remission. Follow-up of immune reconstitution displayed transient severe combined immunodeficiency with slow normalization of the cellular and humoral compartments without a significant increase of infectious complications. Taken together, high dose chemotherapy can be safely given following treatment with CHOP+rituximab. Efficacy in this small cohort of patients was encouraging with sustained remissions in both FCL and MCL patients. Upfront HDT should be considered as a therapeutic option especially in young and/or high-risk patients. PMID- 12732886 TI - Prognostic significance of the detection of tumour cells in peripheral blood stem cell collections in stage II and III breast cancer patients treated with high dose therapy. AB - The purpose of this study was to evaluate the incidence and extent of tumour cell contamination in bone marrow specimens and stem cell collections from 34 breast cancer patients undergoing high-dose therapy as adjuvant treatment, and to determine the prognostic significance for the clinical outcome. Tumour cell contamination was evaluated by flow cytometry using a double-colour test and an anti- Pan cytokeratin (CK) antibody. Two out of 34 (6%) baseline bone marrow specimens, none of seven marrow harvests and nine out of 32 aphereses (28%) mobilised from seven out of 27 patients (26%) contained CK+ cells. Tumour contamination was more frequent in patients with 10 or more involved lymph nodes and in those who received a shorter course of adjuvant chemotherapy before mobilisation. At a median follow-up of 43 months, 24 patients are in complete remission, whereas 10 patients experienced recurrence. Out of the 10 patients who relapsed, five (50%) had CK+ peripheral blood stem cell (PBSC) collections, whereas disease recurrence was seen in only two out of 24 (8%) patients who received CK- products (P=0.02). Moreover, CK+ PBSC collections were associated with a significantly shorter event-free survival and overall survival. CK+ collection is an unfavourable prognostic factor for patients treated with high dose therapy. Whether the negative impact on clinical outcome depends on reinfusion of tumour cells or whether it simply indicates a larger disease extension is still unclear. PMID- 12732885 TI - Cyclosporine is required to prevent severe acute GVHD following T-cell-depleted peripheral blood stem cell transplantation. AB - Reduced immunosuppression may improve immune recovery and increase the graft versus-leukemia effect after allogeneic stem cell transplantation. Furthermore, the requirement for post-transplant immunosuppression following extensive T-cell depletion remains unclear. We therefore evaluated the role of cyclosporine (CSA) in recipients of HLA-identical T-cell-depleted peripheral blood stem cell transplants (PBSCT), followed by donor lymphocyte infusions (DLIs) scheduled on days +45 and +100. Before day+45, successive cohorts of patients received decreasing amounts of CSA: standard-dose (SD) CSA, low-dose (LD) CSA, or no CSA until day+45. LD CSA was as effective as SD CSA in preventing acute graft-versus host disease (GVHD). However, moderate-to-severe acute GVHD was significantly more frequent before the day +45 DLI in patients receiving no CSA (33.3 vs 12.7%, P=0.036, including the only four grade III-IV cases). As a result of higher rates of early acute GVHD, more patients in the 'no CSA' group failed to receive any DLI (30.7 vs 7.1%, P=0.01). Overall, there was no difference in the incidence of acute GVHD, as patients receiving CSA developed more GVHD after DLI. Similarly, no significant differences were found in chronic GVHD, transplant-related mortality, or survival. These results define a role for CSA in preventing GVHD at low T-cell doses following PBSCT. PMID- 12732888 TI - Pre-emptive treatment of CMV DNAemia in paediatric stem cell transplantation: the impact of recipient and donor CMV serostatus on the incidence of CMV disease and CMV-related mortality. AB - Cytomegalovirus (CMV) DNAemia was detected by PCR in 30/125 (24%) consecutive paediatric patients undergoing allogeneic stem cell transplantation. All patients with CMV DNAemia received pre-emptive ganciclovir until two consecutive negative results were obtained. CMV-IgG-positive patients (R+) had a significantly increased risk of DNAemia as compared to CMV-IgG-negative (R-) patients (62% vs 8%) P<0.0001. The incidence of DNAemia was 71% (10/14) in R+ transplanted from seronegative donors (D-) compared to 54% (13/32) in those transplanted from seropositive donors (D+). Of 30 (40%) children with DNAemia, 12 developed CMV disease despite pre-emptive treatment. The overall incidence of disease was 0% (0/59) for R-/D-, 9% (3/23) for R+/D+, 7% (2/29) for R-/D+ and 57% (8/14) for R+/D-. In patients with DNAemia, 4/20 (20%) patients with D+ and 8/10 (80%) with D- became symptomatic. In the multivariate analysis of both groups, patients at risk (R+ and/or D+) and patients with DNAemia, a negative donor serostatus was the only factor associated with a significantly increased incidence of disease. Seven of 9 patients with lethal CMV disease had received CMV-IgG-negative grafts. The data suggest that in CMV seropositive recipients donor CMV seropositivity is associated with a reduced incidence of CMV disease and a favourable outcome following pre-emptive treatment. PMID- 12732887 TI - Surveillance of nosocomial infections in adult recipients of allogeneic and autologous bone marrow and peripheral blood stem-cell transplantation. AB - To identify overall and site-specific rates of nosocomial infections (NIs) during the neutropenic, as compared to the non-neutropenic stage of treatment in adult recipients of allogeneic and autologous bone marrow transplantation (BMT) and peripheral blood stem-cell transplantation (PBSCT), a prospective, 54-month study was started at the Haematological Stem Cell Transplantation Unit of the University Hospital of Freiburg, Germany. NI types were identified using modified CDC definitions. A total of 351 patients (14 256 in-patient days, 5026 neutropenic days) were investigated (316/90% allogeneic, 35/10% autologous; BMT: 119 patients, PBSCT: 234 patients). The mean length of neutropenia was 14.3 days (range: 0-66). Antimicrobial prophylaxis for allogeneic transplantation consisted of ciprofloxacin, trimethoprim/sulpha-methoxazole, fluconazole, and metronidazole. In total, 239 NIs were identified in 169 patients (48.1%), and of these 171 (71.5%) occurred during neutropenia (34.0 NIs per 1000 days at risk). The main pathogens were coagulase-negative staphylococci (36.3%), Clostridium difficile (20.4%), and enterococci (10.0%). Site-specific incidence densities during neutropenia vs non-neutropenia were: 13.9 vs 1.6 bloodstream infections (all central line-associated), 11.9 vs 1.8 pneumonias, 3.0 vs 2.9 gastroenteritis, and 1.6 vs 0.3 urinary tract infections. The greatest number of NI in BMT and PBSCT recipients is acquired during neutropenia, and multicentre surveillance programmes should focus on this. PMID- 12732889 TI - Loss of hepatitis A virus (HAV) antibodies after peripheral stem cell transplantation (PSCT). AB - The majority of patients with hepatitis A have a benign course, but some may develop fulminant hepatitis and hematological complications. Peripheral stem cell transplantation (PSCT) is associated with loss of immunity. There are no data regarding loss of HAV antibodies (anti-HAV) after PSCT. We retrospectively evaluated the persistence of anti-HAV in a nonvaccinated population that underwent PSCT. Serum detection of anti-HAV was determined before and after PSCT using a qualitative commercially available enzyme immunoassay. From January 1997 to March 2001, 136 (68%) of 201 patients tested (+) for anti-HAV prior to PSCT. Subsequent investigation of anti-HAV was possible in 36 of these patients at a median of 12 months after PSCT. The median age of patients was 47 years old; they had diagnoses of hematological malignancies (33) and solid tumors (three), and underwent autologous (31) and allogenic (five) PSCT. A total of 31 (86%) of 36 patients remained anti-HAV (+) and five (14%) became (-) after PSCT. The variables age, sex, diagnosis, type of PSCT, time of testing, and number of CD34 cells infused were not predictors of loss of anti-HAV. In conclusion, 14% of 36 nonvaccinated anti-HAV (+) patients lost their antibodies at a median of 12 months after PSCT. PMID- 12732890 TI - Low incidence of CMV viremia and disease after allogeneic peripheral blood stem cell transplantation. Role of pretransplant ganciclovir and post-transplant acyclovir. AB - To establish the incidence of CMV viremia after allogeneic blood stem cell transplantation, we studied 51 consecutive allogeneic peripheral blood stem cell (PBSC) transplant recipients. A total of 12 recipients were at moderate risk for CMV disease and 39 were at high risk. Conditioning regimens varied, but GvHD prophylaxis consisted of tacrolimus and mini-methotrexate in all patients. All patients received prophylactic ganciclovir from admission until day -2 and prophylactic acyclovir from day -1 until day 180 after transplantation. CMV viremia was treated with ganciclovir. Using a PCR-based technique, the cumulative incidence of CMV viremia was 31+/-14% by day 100 and 35+/-14% by day 150. Donor type, CMV risk group, underlying disorder, conditioning regimen, GvHD, and steroid use were not associated with the risk for CMV viremia. No cases of CMV disease occurred. We hypothesize that the low rate of CMV viremia and the absence of CMV disease in this cohort of PBSCT transplant recipients, which contrasts with other reports, may be related to the prophylactic use of high-dose acyclovir and possibly to pretransplant use of ganciclovir. PMID- 12732891 TI - Non-overt disseminated intravascular coagulation in patients during treatment with antithymocyte globulin for unrelated allogeneic hematopoietic stem cell transplantation. AB - We assessed the effect of rabbit antithymocyte globulin manufactured by Fresenius (ATG-F) on the hemostatic system in patients (n=12) with various hematological malignancies undergoing hematopoietic stem cell transplantation (HSCT) from HLA matched unrelated donors. For this purpose, we monitored different parameters of coagulation before, during and after the administration of ATG-F. As a control group, we recruited patients (n=10) undergoing HSCT from their HLA-identical siblings who did not receive ATG-F as part of their preparative regimens. At 24 and 48 h after ATG-F treatment had been initiated, we found a temporary rise in D Dimer, tissue factor, soluble thrombomodulin and thrombin-antithrombin III complex levels and a significant decrease of platelet counts in patients treated with ATG-F as compared to the control group. No differences between the two groups could be detected with regard to global coagulation tests as well as the incidence of bleeding manifestations, thromboembolic complications or the development of vascular-occlusive-disease of the liver. This temporary state of a stressed but compensated coagulation system under ATG-F therapy can be addressed as nonovert disseminated intravascular coagulation (DIC). The effect was independent from the different conditioning regimens and eased off after cessation of ATG-F. We conclude that ATG-F can induce nonovert DIC in patients receiving antithymocyte globulin as part of their conditioning regimen for HSCT. PMID- 12732892 TI - Preclinical evaluation of an automated closed fluid management device: Cytomate, for washing out DMSO from hematopoietic stem cell grafts after thawing. AB - Infusion of dimethylsulfoxide (DMSO) contained in cryopreserved and thawed hematopoietic stem cell (HSC) grafts is frequently associated with mild or moderate adverse reactions, and occasionally with more severe events including neurological symptoms. The severity of these complications is related to the amount of residual DMSO. We evaluated a recently available, closed, automated and 'cgmp (current good manufacturing practice) compliant' device (CytoMate) for its ability to wash out DMSO at the expense of a limited loss of viable CD34(+) cells. A total of 16 procedures were carried out with 39 blood HSC bags intended for destruction. Mean amounts of DMSO for each cellular product (one, two or three bags) were between 12.2 and 39.6 g before thawing; after the washing procedure, residual DMSO quantities were between 0.1 and 3.7 g. When set up to reproducibly allow for a more than 96% elimination of DMSO, processing of thawed cells with the CytoMate cell processor resulted in a mean recovery of viable total cells, CD34(+) cells and lymphocyte subsets above 60%. We conclude that this simple and efficient washing technique is suitable for routine processing of HSC grafts. Clinical studies will demonstrate whether a reduction in the incidence of adverse effects associated with DMSO infusion is observed. PMID- 12732893 TI - Autoimmune hepatitis following allogeneic PBSCT from an HLA-matched sibling. AB - A 7-year-old boy with acute lymphoblastic leukemia (ALL) in second remission received an allogeneic PBSCT from his HLA-matched sister. Acute grade II graft versus-host disease (GVHD) resolved with corticosteroids. Chronic GVHD in the skin and oral mucosa at around day 60 responded to corticosteroids and cyclosporin A. At 6 months after the transplant, he developed hepatic dysfunction with elevated serum transaminases and gamma-globulin. Liver biopsy revealed chronic inflammation with lymphocytes and plasma cells in portal areas without destruction of bile ducts, suggesting autoimmune hepatitis. While rare, autoimmune hepatitis should be considered a potential long-term complication in patients with hepatic dysfunction in the late post-transplant phase. PMID- 12732894 TI - Imminent allograft rejection prevented by donor lymphocyte transfusions: report of two pediatric cases. AB - Gradual allograft rejection after initial good engraftment may occur with simultaneous autologous reconstitution particularly in patients receiving nonmyeloablative conditioning. Careful post-transplant follow-up of the chimerism status can reveal these cases early on, when the immunological balance may still be shifted to the donor cells. We describe two children with nonmalignant diseases, in whom imminent rejection of their sibling allografts was prevented with donor lymphocyte transfusions (DLT). DLT dosing and timing need to be individually guided by monitoring of the chimerism status. PMID- 12732897 TI - NTP Toxicology and Carcinogenesis Studies of Xylenes (Mixed) (60% m-Xylene, 14% p Xylene, 9% o-Xylene, and 17% Ethylbenzene) (CAS No. 1330-20-7) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - The technical grade of xylenes (mixed) (hereafter termed xylenes) contains the three isomeric forms and ethylbenzene (percentage composition shown above). The annual production for 1985 was approximately 7.4 x 108 gallons. Xylenes is used as a solvent and a cleaning agent and as a degreaser and is a constituent of aviation and automobile fuels. Xylenes is also used in the production of benzoic acid, phthalate anhydride, and isophthalic and terephthalic acids as well as their dimethyl esters. Toxicology and carcinogenesis studies of xylenes were conducted in laboratory animals because a large number of workers are exposed and because the long- term effects of exposure to xylenes were not known. Exposure for the present studies was by gavage in corn oil. In single-administration studies, groups of five F344/N rats and B6C3F1 mice of each sex received 500, 1,000, 2,000, 4,000, or 6,000 mg/kg. Administration of xylenes caused deaths at 6,000 mg/kg in rats and mice of each sex and at 4,000 mg/kg in male rats. In rats, clinical signs observed within 24 hours of dosing at 4,000 mg/kg included prostration, muscular incoordination, and loss of hind limb movement; these effects continued through the second week of observation. Tremors, prone position, and slowed breathing were recorded for mice on day 3, but all mice appeared normal by the end of the 2- week observation period. In 14- day studies, groups of five rats of each sex were administered 0, 125, 250, 500, 1,000, or 2,000 mg/kg, and groups of five mice of each sex received 0, 250, 500, 1,000, 2,000, or 4,000 mg/kg. Chemical- related mortality occurred only at 2,000 mg/kg in rats and at 4,000 mg/kg in mice. Rats and mice exhibited shallow breathing and prostration within 48 hours following dosing at 2,000 mg/kg. These signs persisted until day 12 for rats, but no clinical signs were noted during the second week for mice. In 13- week studies, groups of 10 rats of each sex received 0, 62.5, 125, 250, 500, or 1,000 mg/kg, and groups of 10 mice of each sex received 0, 125, 250, 500, 1,000, or 2,000 mg/kg. No deaths or clinical signs of toxicity were recorded in rats. However, high dose male rats gained 15% less weight and females 8% less weight than did the vehicle controls. Two female mice died at the 2,000 mg/kg dose. Lethargy, short and shallow breathing, unsteadiness, tremors, and paresis were observed for both sexes in the 2,000 mg/kg group within 5- 10 minutes after dosing and lasted for 15- 60 minutes. Two- year toxicology and carcinogenesis studies were conducted by administering 0, 250, or 500 mg/kg xylenes in corn oil by gavage to groups of 50 F344/N rats of each sex, 5 days per week for 103 weeks. Groups of 50 B6C3F1 mice of each sex were administered 0, 500, or 1,000 mg/kg xylenes on the same schedule. Although the mortality was dose related in male rats (final survival: vehicle control, 36/50; low dose, 26/50; high dose, 20/50), many of the early deaths in the dosed males were gavage related. Body weights of the high dose male rats were 5%- 8% lower than those of the vehicle controls after week 59. The mean body weights of low dose and vehicle control male rats and those of dosed and vehicle control female rats were comparable. Survival rates of female rats and both sexes of dosed mice were not significantly different from those of the vehicle controls. The mean weights of dosed male and female mice were comparable to those of the vehicle controls. Hyperactivity lasting 5- 30 minutes was observed in high dose mice after dosing, beginning after week 4 and continuing through week 103. At no site was the incidence of nonneoplastic or neoplastic lesions in dosed rats or mice of either sex considered to be related to the administration of xylenes. Neither xylenes nor any of its components (o- xylene, m-xylene, p- xylene, or ethylbenzene) were mutagenic when tested with or without metabolic activation in Salmonella typhimurium strains TA100, TA1535, TA97, or TA98 with the preincubation protocol. In addition, ethylbenzene was tested in cytogenetic assays using cultured Cetic assays using cultured Chinese hamster ovary cells both with and without metabolic activation; neither sister- chromatid exchanges nor chromosomal aberrations were induced by ethylbenzene. An audit of the experimental data was conducted for the 2-year studies of xylenes. No data discrepancies were found that influenced the final interpretations. Under the conditions of these 2-year gavage studies, there was no evidence of carcinogenicity of xylenes (mixed) for male or female F344/N rats given 250 or 500 mg/kg or for male or female B6C3F1 mice given 500 or 1,000 mg/kg. PMID- 12732898 TI - NTP Toxicology and Carcinogenesis Studies of Erythromycin Stearate (CAS No. 643 22-1) in F344/N Rats and B6C3F1 Mice (Feed Studies). AB - Toxicology and carcinogenesis studies of erythromycin stearate (USP grade, greater than 96% pure) were conducted by administering the antibiotic in feed to groups of F344/N rats and B6C3F1 mice of each sex for 14 days, 13 weeks, or 2 years. Erythromycin stearate was studied because of its widespread use in humans as a broad-spectrum macrolide antibiotic and because of the lack of adequate long term studies for carcinogenicity. Fourteen-Day and Thirteen-Week Studies: In the 14-day studies, none of the rats (at dietary concentrations up to 50,000 ppm) and 2/5 female mice that received 50,000 ppm died before the end of the studies. Final mean body weights of male rats that received 12,500, 25,000, or 50,000 ppm were 10%, 30%, or 36% lower, respectively, than that of controls; final mean body weights of female rats were 10%, 12%, or 32% lower. None of the dosed mouse groups gained weight. The final mean body weight of male mice that received 50,000 ppm was 10% lower than that of controls. In the 13-week studies, none of the rats or mice (at dietary concentrations up to 20,000 ppm) died before the end of the studies. Final mean body weights of the 20,000-ppm groups of rats were more than 12% lower than that of the controls for males and 7% lower for females. Final mean body weights of mice that received 10,000 or 20,000 ppm were 15% or 19% lower than that of controls for males and 5% or 14% lower for females. Multinucleated syncytial hepatocytes were observed in 10/10 male rats that received 20,000 ppm but in 0/10 male rats that received 10,000 ppm. No compound related gross or microscopic pathologic effects were observed in mice. Based on these results, 2-year studies of erythromycin stearate were conducted by feeding diets containing 0, 5,000, or 10,000 ppm erythromycin stearate to groups of 50 rats of each sex for 103 weeks. Diets containing 0, 2,500, or 5,000 ppm were fed to groups of 50 mice of each sex for 103 weeks. Body Weight and Survival in the Two-Year Studies: Mean body weights of high dose male rats were comparable to those of controls throughout the studies. Mean body weights of high dose female rats were 5%-10% lower than those of controls. Mean body weights of dosed and control mice were comparable. The average daily feed consumption was similar for dosed and control male and female rats. For mice, estimated daily feed consumption by low and high dose males was similar to that of the controls and by low and high dose females was 92% that of the controls. The average amount of erythromycin stearate consumed per day was approximately 180 or 370 mg/kg for male rats and 210 or 435 mg/kg for female rats; for mice, the average amounts were 270 or 545 mg/kg for males and 250 or 500 mg/kg for females. No significant differences in survival were observed between any groups of rats or mice of either sex (final survival-- male rats: control, 28/50; low dose, 23/50; high dose, 27/50; female rats: 29/50; 30/50; 38/50; male mice: 34/50; 33/50; 40/50; female mice: 38/50; 34/50; 40/50). Nonneoplastic and Neoplastic Effects in the Two-Year Studies: Granulomas of the liver were observed at increased incidences in high dose rats (male: 1/50; 2/50; 10/50; female: 18/50; 27/50; 43/50). Granulomatous inflammation or granulomas of the spleen were observed in dosed female rats (0/48; 1/49; 3/50). Reticulum cell hyperplasia in the bone marrow occurred at increased incidences in high dose female rats (10/50; 14/50; 25/50). Squamous cell papillomas of the oral mucosa were observed in 1/50 control, 2/50 low dose, and 3/50 high dose female rats. These tumors were considered to be marginal and not related to exposure. Hyperplasia of the oral mucosa was not observed. Pheochromocytomas of the adrenal gland in female rats occurred with a positive trend (1/50; 4/49; 5/50). The incidences in the dosed groups are similar to the average historical incidence (9%) of this tumor in untreated control female F344/N rats at the study laboratory. This marginal tumor increase is not considered to be biologically important. No increases in incidences of study laboratory. This marginal tumor increase is not considered to be biologically important. No increases in incidences of neoplasms were observed at any site in dosed male rats. Inflammation in the glandular stomach was observed at increased incidences in dosed male mice (1/49; 4/50; 6/50). Lymphoid hyperplasia in the urinary bladder was observed at increased incidences in dosed female mice (1/50; 9/47; 7/48). No increases in incidences of neoplasms were observed at any site in dosed male or female mice. Genetic Toxicology: Erythromycin stearate was not mutagenic in Salmonella typhimurium strains TA98, TA100, TA1535, or TA1537 when tested both with or without exogenous metabolic activation. Erythromycin stearate demonstrated equivocal mutagenicity in the mouse L5178Y lymphoma cell assay in the absence of exogenous metabolic activation (S9); erythromycin stearate was not mutagenic in the presence of S9. Treatment of cultured Chinese hamster ovary cells with erythromycin stearate did not produce an increase in the frequency of sister chromatid exchanges or chromosomal aberrations in either the presence or absence of metabolic activation. Audit: The data, documents, and pathology materials from the 2-year studies of erythromycin stearate have been audited. The audit findings show that the conduct of the studies is documented adequately and support the data and results given in this Technical Report. Conclusions: Under the conditions of these 2-year studies, there was no evidence of carcinogenic activity of erythromycin stearate for male or female F344/N rats administered erythromycin stearate in the diet at 5,000 or 10,000 ppm. There was no evidence of carcinogenic activity of erythromycin stearate for male or female B6C3F1 mice administered erythromycin stearate in the diet at 2,500 or 5,000 ppm. Dose related increases in the incidences of granulomas of the liver were observed in male and female rats. The absence of any biologically important chemical associated effects in mice suggests that higher doses could have been given to male and female mice. Synonyms: erythrocin stearate; erythromycin octadecanoate Trade Names: Abbotcine; Bristamycin; Dowmycin E; Eratrex; Erypar; Ethril; Gallimycin; HSDB 4178; OE 7; Pantomicina; Pfizer-E; SK-Erythromycin; Wyamycin S PMID- 12732899 TI - NTP Toxicology and Carcinogenesis Studies of Nitrofurazone (CAS No. 59-87-0) in F344/N Rats and B6C3F1 Mice (Feed Studies). AB - Nitrofurazone is a synthetic furan derivative, active against a broad spectrum of bacteria, which has been widely used in veterinary and human medicine. Toxicology and carcinogenesis studies were conducted by feeding diets containing nitrofurazone (99% pure) to groups of F344/N rats and B6C3F1 mice of each sex for 14 days, 13 weeks, or 2 years. Fourteen-Day and Thirteen-Week Studies: Groups of five males and five females of each species were fed diets containing 0, 630, 1,250, 2,500, 5,000, or 10,000 ppm for 14 consecutive days. Early deaths occurred in all groups of rats receiving 5,000 or 10,000 ppm nitrofurazone. The surviving rats in the lower two dose groups gained weight, but weight gain was decreased as the dose of nitrofurazone was increased. Feed consumption by rats of each sex was decreased at all doses above 630 ppm. In all dosed groups, clinical signs of toxicity included rough hair coats and lethargy. At doses of 2,500 ppm and above, rats of each sex exhibited intermittent episodes of seizures and lethargy. All mice that received 2,500, 5,000, or 10,000 ppm nitrofurazone and 3/5 males that received 1,250 ppm died before the end of the 14-day studies; the surviving dosed mice (except females at 630 ppm) lost weight. A dose-related decrease in feed consumption was observed at all doses above 630 ppm. Clinical signs included rough hair coats and convulsive seizures. In the 13-week studies, groups of 10 rats of each sex were given diets containing 0, 150, 310, 620, 1,250, or 2,500 ppm nitrofurazone. No deaths were observed and all animals gained weight, but the magnitude of weight gain was dose dependent with decrements in final mean body weight for the highest dose group reaching 55% in males and 36% in females. Other evidence of chemically related toxicity included convulsive seizures, osteoporosis, degenerative arthropathy, and gonadal hypoplasia in both sexes at the two highest doses. Groups of 10 mice of each sex were given diets containing 0, 70, 150, 310, 620, or 1,250 ppm nitrofurazone for 13 weeks. Early deaths were observed in the two highest dose groups of each sex. The final mean body weights of male and female mice in the 1,250-ppm groups were about 20% lower than those of the controls; weight gains of the other dosed mice were comparable to those of the controls. Stimulus-induced convulsive seizures were observed for all mice in the two highest dose groups. Testicular hypoplasia was observed in the two highest dose groups of male mice. Body Weight and Survival in the Two-Year Studies: Dietary concentrations for the 2-year studies were 0, 310, or 620 ppm for rats and 0, 150, or 310 ppm for mice (50 animals per dose group). Mean body weights of high dose male rats were lower than those of the controls after week 39; mean body weights of low dose male rats and of the controls were comparable throughout the study. Final mean body weights of low and high dose female rats were 9% and 21% lower than those of the controls. Dosed rats consumed less feed than did the controls. The average amount of nitrofurazone consumed per day was approximately 11-12 or 24-26 mg/kg by low or high dose male and female rats. The survival of the high dose group of male rats was lower than that of the controls after week 92 (final survival-- male: control, 33/50; low dose, 30/50; high dose, 20/50; female: 28/50; 37/50; 31/50). Mean body weights of dosed mice were similar to or somewhat greater than those of controls throughout most of the studies. The average daily feed consumption by dosed mice was similar to that of controls. The average amount of nitrofurazone consumed per day was approximately 14-16 or 29-33 mg/kg for low or high dose male and female mice. The survival of the high dose group of male mice was lower than that of the controls after week 88 (final survival-- male: 39/50; 31/50; 27/50; female: 39/50; 40/50; 35/50). In mice of each sex, nitrofurazone administration induced stimulus-sensitive convulsive seizures beginning at week 4 or 5 for high dose mice and week 24 for low dose female mice. These seizures were low dose female mice. These seizures were observed primarily in the first year of the study. Nonneoplastic and Neoplastic Effects in the Two-Year Studies: Degenerative changes involving the vertebral and femoro-tibial (knee) joints were observed at increased incidences in dosed rats. The degenerative changes primarily affected the articular cartilage and were similar to those seen in the 13-week studies. Degeneration of the sternal synchondroses was increased in high dose female rats. The osteoporosis seen in the 13-week studies was not observed in the 2-year studies. Testicular degeneration, characterized by atrophy of the germinal epithelium and aspermatogenesis, was observed at increased incidences in dosed male rats (control, 12/50; low dose, 49/50; high dose, 47/50). Adenomas of the sebaceous glands and trichoepitheliomas or sebaceous adenomas (combined) of the skin were observed in high dose male rats (0/50; 0/50; 5/50). Carcinomas of the preputial gland were increased in dosed male rats (1/50; 8/50; 5/50). The incidences of preputial gland adenomas or carcinomas (combined) in dosed male rats were not statistically greater than that in the controls (9/50; 16/50; 7/50). However, in the low dose group, the incidence is greater than the highest incidence observed in historical untreated control groups (9/50). In addition, hyperplasia of the preputial gland was observed in six low dose male rats in which neither adenomas nor carcinomas occurred. The incidence of mesotheliomas of the tunica vaginalis in low dose male rats was greater than that in the controls (0/50; 7/50; 2/50). Fibroadenomas of the mammary gland occurred at markedly increased incidences in dosed female rats (8/49; 36/50; 36/50). Three adenocarcinomas were also observed (1/49; 0/50; 2/50). Ovarian atrophy (7/47; 44/50; 38/50) and tubular cell hyperplasia of the ovary (1/47; 23/50; 21/50) were observed at markedly increased incidences in dosed female mice. The incidences of benign mixed tumors (0/47; 17/50; 20/50), granulosa cell tumors (1/47; 4/50; 9/50), and granulosa cell tumors or luteomas (combined) (3/47; 6/50; 9/50) of the ovary were increased in exposed female mice. Mononuclear cell leukemia in rats occurred with negative trends (male: 21/50; 23/50; 6/50; female: 15/49; 2/50; 2/50). In female mice, the incidences of adenomas or carcinomas (combined) of the anterior pituitary gland occurred with a negative trend (10/50; 7/50; 2/49). The incidences of testicular interstitial cell tumors were decreased in dosed male rats (45/50; 30/50; 28/50). Genetic Toxicity: Nitrofurazone was mutagenic in Salmonella typhimurium strains TA98 and TA100 both with and without exogenous metabolic activation. The responses in strains TA1535 and TA1537 were more varied: nitrofurazone was mutagenic in strain TA1535 only in the presence of S9 and produced no consistent increase in gene reversions in strain TA1537 with or without S9. In the absence of metabolic activation, nitrofurazone induced forward mutations at the TK+/- locus of mouse L5178Y lymphoma cells; the chemical was not tested with S9. Treatment of cultured Chinese hamster ovary cells with nitrofurazone in the absence of S9 produced a dose-related increase in sister chromatid exchanges and chromosomal aberrations; with S9, sister chromatid exchanges were increased, but no induction of chromosomal aberrations was observed. Audit: The data, documents, and pathology materials from the 2-year studies of nitrofurazone were audited at the NTP Archives. The audit findings show that the conduct of the studies is documented adequately and support the data and results given in this Technical Report. Conclusions: Under the conditions of these 2-year feed studies, there was equivocal evidence of carcinogenic activity of nitrofurazone for male F344/N rats as shown by the occurrence of sebaceous gland adenomas and trichoepitheliomas of the skin, mesotheliomas of the tunica vaginalis, and preputial gland tumors. There was clear evidence of carcinogenic activity of nitrofurazone for female F344/N rats as shown by a markedly increased incidence of fibroadenomas of the mammary gland. There was no evidence of carcinogenic activity for male B6C3F1 mice fed diets containing nitrofurazone at concentrations of 150 or 310 ppm. There was clear evidence of carcinogenic activity of nitrofurazone for female B6C3F1 mice as shown by increased incidences of benign mixed tumors and granulosa cell tumors of the ovary. Administration of nitrofurazone was associated with decreased incidences of mononuclear cell leukemia in male and female rats, testicular interstitial cell tumors in male rats, and pituitary gland neoplasms in female mice. Convulsive seizures in mice of each sex, ovarian atrophy in female mice, testicular degeneration in rats, and degeneration of articular cartilage in rats were all associated with the administration of nitrofurazone. Synonyms: 5-nitro-2-furaldehyde semicarbazone; 2-[(5-nitro-2 furanyl)methylene]hydrazine carboximide Trade Names: Aldomycin; Amifur; Chemfuran; Coxistat; Furacin; Furacinetten; Furaplast; Furazol W; Furesol; Furracoccid; Mammex; Nefco; Nifuzon; Nitrofural; Vabrocid PMID- 12732900 TI - NTP Toxicology and Carcinogenesis Studies of Penicillin VK (CAS No. 132-98-9) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - Penicillin VK, a widely used antibiotic for treatment of gram-positive coccal infections, was nominated for study by the National Cancer Institute because rodent carcinogenicity studies for this drug had not been performed. The chemical (94% or 98% pure, USP grade) was administered orally (by gavage in corn oil) because oral administration is the primary route used to treat infections in humans. Fourteen-day, 13-week, and 2-year studies were conducted in F344/N rats and B6C3F1 mice. Additional studies were performed to evaluate the potential for genetic damage in bacteria and mammalian cells. Fourteen-Day and Thirteen-Week Studies: In the 14-day studies, penicillin VK was administered at doses of 150 2,400 mg/kg. No compound-related deaths or dose-related histopathologic lesions were seen in rats or mice. Final mean body weights of dosed male rats were 5%-17% lower than that of controls; weights of dosed and control female rats were comparable. Final mean body weights of dosed mice were 5%-9% lower than those of controls. Diarrhea was observed in all dosed groups of rats and mice. In the 13 week studies, male and female rats received doses of 180-3,000 mg/kg and male and female mice received doses of 250-3,000 mg/kg. No compound-related deaths were seen in rats or mice. Final mean body weights of rats that received 3,000 mg/kg were 11% lower than those of the vehicle controls for males and 6% lower for females. For mice, mean body weights were comparable. Diarrhea occurred in male rats at doses of 750 mg/kg and above and in female rats at doses of 1,500 and 3,000 mg/kg. Mucous cell metaplasia of the glandular stomach was observed in male and female rats receiving 1,500 and 3,000 mg/kg. Lesions of the glandular stomach (inflammation, mucous cell metaplasia, and eosinophilic cytoplasmic change) and the forestomach (papillary hyperplasia and hyperkeratosis) were seen in all groups of dosed mice. The severity of lesions at 1,000 mg/kg or below was considered minimal. Based on these results, doses selected for rats and mice in the 2-year studies were 0, 500, or 1,000 mg/kg. Body Weight and Survival in the Two-Year Studies: Mean body weights of dosed and vehicle control male and female rats and male mice were comparable. Mean body weights of dosed female mice were 4%-16% lower than those of the vehicle controls from week 28 to the end of the study. Diarrhea was observed for dosed male and female rats and for dosed male mice. Survival of low and high dose male rats and high dose female rats was reduced (male rats: vehicle control, 34/50; low dose, 19/50; high dose, 16/50;female rats: 29/50; 26/50; 16/50). Survival of male and female mice was comparable to that of the vehicle controls (male mice: 24/50; 36/50; 26/50; female mice: 36/50; 32/50; 32/50). Nonneoplastic and Neoplastic Effects in the Two-Year Studies: Nonneoplastic lesions occurred at low incidences in the nasal mucosa, lung, and forestomach of dosed male rats and in the nasal mucosa and lung of dosed female rats. Congestion and aspiration pneumonia occurring in dosed rats dying before week 104 was the principal cause of death in these animals. Nonneoplastic lesions of the gastric fundal gland (eosinophilic cytoplasmic change and dilatation) and glandular stomach (cyst, chronic focal inflammation, hyperplasia, fibrosis, and squamous metaplasia) were seen in dosed male and female mice, and lesions of the gallbladder (eosinophilic cytoplasmic change) were seen in male mice. Slight increases in the incidences of adenomas of the pituitary gland in high dose male rats and of fibroadenomas or adenomas (combined) of the mammary gland in low dose female rats were observed. These were not considered to be compound-related lesions. The incidence of hepatocellular adenomas was decreased in high dose male mice (14/50; 15/49; 4/49). No compound related neoplasms were seen in female mice. Genetic Toxicology: Penicillin VK was not mutagenic in Salmonella typhimurium strains TA98, TA100, TA1535, or TA1537 with or without exogenous metabolic activation. The chemical was mutagenic onl exogenous metabolic activation. The chemical was mutagenic only with activation in the mouse lymphoma L5178Y/TK± forward mutation assay. Incubation of Chinese hamster ovary cells with penicillin VK resulted in increased frequencies of sister chromatid exchanges and chromosomal aberrations in the absence of metabolic activation under the conditions of delayed harvest to compensate for chemical-induced cell cycle delay, no effects from penicillin VK exposure were observed in these cells in the presence of S9. Audit: The data, documents, and pathology materials from the 2-year studies of penicillin VK were audited. The audit findings show that the conduct of the studies is documented and support the data and results given in this Technical Report. Conclusions: Under the conditions of these 2-year gavage studies, there was no evidence of carcinogenic activity of penicillin VK for F344/N rats or for B6C3F1 mice administered 500 or 1,000 mg/kg penicillin VK in corn oil gavage, 5 days per week for 2 years. Nonneoplastic lesions were seen in the glandular stomach of dosed mice. Decreased survival of low and high dose male rats and of high dose female rats reduced the sensitivity of the studies for determining the presence or absence of a carcinogenic response in this species. Synonyms: 4-thia-1 azabicyclo(3.2.0)heptane-2-carboxylic acid, 3,3-dimethyl-7-oxo-6-(2-phenoxy acetamide)-, monopotassium salt; penicillin V potassium; penicillin V potassium salt; D-a-phenoxymethylpenicillinate K salt; phenoxymethylpenicillin potassium; PVK Trade Names: Antibiocin; Apsin VK; Aracil; Arcasin; Aspin VK; Beromycin; Beromycin 400; Betapen VK; Calciopen K; Cliacil; Compocillin VK; Distakaps V-K; Distaquaine V-K; Dowpen V-K; DQV-K; Fenoxypen; Icipen; Isocillin; Ispenoral; Ledercillin VK; Megacillin oral; Oracil-VK; Orapen; Ospeneff; Pedipen; Penagen; Pencompren; Pen-Vee K; Pen-V-K powder; Penvikal; Pfizerpen VK; Qidpen VK; Robicillin VK; Rocillin-VK; Roscopenin; SK-Penicillin VK; Stabilin VK Syrup 125; Stabilin VK Syrup 62.5; Sumapen VK; Suspen; Uticillin VK; V-Cil-K; V-Cillin K; Veetids; Vepen PMID- 12732901 TI - NTP Toxicology and Carcinogenesis Studies of C.I. Acid Orange 3 (CAS No. 6373-74 6) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - C.I. Acid Orange 3 is a dinitrodiphenylamine derivative used exclusively as a dye (up to 0.2%) in semipermanent hair coloring products. This study was one of a series on semipermanent hair dyes, which included HC Blue No. 1 (NTP TR 271), HC Blue No. 2 (NTP TR 293), HC Red No. 3 (NTP TR 281), and C.I. Disperse Blue 1 (NTP TR 299). Toxicology and carcinogenesis studies of C.I. Acid Orange 3 (90% pure, containing 10% water for short-term studies and containing 6%-8% water and 2%-4% acetone for 2-year studies) were conducted by administering the dye in corn oil by gavage to F344/N rats and B6C3F1 mice of each sex for 14 days, 13 weeks, or 2 years. Fourteen-Day and Thirteen-Week Studies: In the 14-day studies (at 94-1,500 mg/kg in rats and 62-1,000 mg/kg in mice), no compound-related deaths or body weight changes were observed and no adverse effects were observed at necropsy. In the 13-week studies (at 94-1,500 mg/kg in rats and 31-2,000 mg/kg in mice), compound-related kidney lesions were observed in rats and mice of each sex. These lesions included variable degrees of degeneration and necrosis of epithelial cells in the proximal convoluted tubules, regeneration of tubular epithelium, and granular casts in the tubules. In a few female rats of the highest dose group, necrosis of the renal papillae and suppurative inflammation were also observed. Mean body weights were generally comparable among groups of rats and mice. Mice receiving 2,000 mg/kg had body weights 11%-12% lower than those of vehicle controls. Five of 10 female rats that received the highest dose of 1,500 mg/kg died before the end of the study, but no compound-related deaths occurred in male rats or mice of either sex. Based on these results, 2-year studies of C.I. Acid Orange 3 were conducted by administering the dye by gavage in corn oil at 0, 375, or 750 mg/kg to groups of 50 F344/N rats of each sex, 5 days per week for 103 weeks. Groups of 50 male B6C3F1 mice were administered 0, 125, or 250 mg/kg C.I. Acid Orange 3 on the same schedule, and groups of 50 female B6C3F1 mice were administered 0, 250, or 500 mg/kg. These doses were selected on the basis of the nature and severity of the renal lesions in both species. Body Weights and Survival in the Two-Year Studies: Mean body weights of high dose rats were generally more than 10% lower than those of vehicle controls after week 52 for males and week 70 for females. Mean body weights forlow dose groups were comparable to those of vehicle controls. The survival of high dose male (after week 33) and female (after week 14) rats was lower (P<0.05) than that of vehicle controls and was attributed to nephrotoxicity (final survival-- male: vehicle control, 36/50; low dose, 30/50; high dose, 0/50; female: 43/50; 34/50; 7/50). Mean body weights of dosed male and female mice were lower than those of vehicle controls (high dose, 5%-11% after week 74; low dose, 7%-17% after week 48). Survival of both the low dose (after week 102) and high dose (after week 100) groups of male mice was lower than that of the vehicle controls (final survival: 38/50; 25/50; 26/50). Although survival was lower than usual, no notable differences in survival were observed between groups of female mice (final survival: 23/50; 23/50; 24/50). Nonneoplastic and Neoplastic Lesions in the Two Year Studies: For both species, the kidney was the major target organ for C.I. Acid Orange 3. These findings are summarized in the accompanying table. The incidences of renal pelvic epithelial hyperplasia were increased in dosed rats of each sex. No renal neoplasms were observed in dosed male rats, but a tubular cell adenocarcinoma was observed in a vehicle control male rat. Six transitional cell carcinomas of the kidney were observed in high dose female rats; kidney transitional cell neoplasms have not been observed in 1,697 corn oil vehicle control female F344/N rats. Nonneoplastic lesions characteristic of secondary renal hyperparathyroidism or secondary to uremia also occurred in dosed rats. The lesions included parathyroid hyperplasia, fibrous dysplasia of boia also occurred in dosed rats. The lesions included parathyroid hyperplasia, fibrous dysplasia of bone, erosion and ulcers of the glandular stomach, and mineralization of the aorta and glandular stomach. Epithelial hyperplasia of the urinary bladder was observed in one low dose and three high dose female mice. A squamous cell carcinoma was seen in the urinary bladder of one low dose female mouse. Even though no squamous cell urinary bladder neoplasms have been observed in 1,665 corn oil vehicle control female B6C3F1 mice, this single neoplasm in a low dose animal was not considered to be related to the administration of C.I. Acid Orange 3. Genetic Toxicology: C.I. Acid Orange 3 was mutagenic with and without exogenous metabolic activation in Salmonella typhimurium strains TA97; TA98; and TA100; no mutagenicity was observed for strain TA1535. Audit: The data, documents, and pathology materials from the 2-year studies of C.I. Acid Orange 3 have been audited. The audit findings show that the conduct of the studies is documented adequately and support the data and results given in this Technical Report. Conclusions: Under the conditions of these 2-year gavage studies, there was no evidence of carcinogenic activity of C.I. Acid Orange 3 for male F344/N rats administered 375 mg/kg; because of a marked reduction in survival and no indication of carcinogenicity, the 750 mg/kg group was considered to be inadequate for assessment of carcinogenic activity. There was clear evidence of carcinogenic activity of C.I. Acid Orange 3 for female F344/N rats as shown by the occurrence of transitional cell carcinomas of the kidney in the 750 mg/kg group; this group had reduced survival and chemically related nonneoplastic lesions of the kidney. There was no evidence of carcinogenic activity of C.I. Acid Orange 3 for male B6C3F1 mice administered 125 or 250 mg/kg or for female B6C3F1 mice administered 250 or 500 mg/kg. Nonneoplastic lesions of the kidney were observed in both dose groups of both sexes of rats and mice. Synonyms: 2 anilino-5-(2,4-dinitroanilino)-benzenesulfonic acid, monosodium salt; 5[(2,4 dinitrophenol)amine]-2-(phenylamine)-benzenesulfonic acid, monosodium salt; C.I. 10385; Tetracid Light Yellow 2R PMID- 12732902 TI - NTP Toxicology and Carcinogenesis Studies of 2-Amino-5-Nitrophenol (CAS No. 121 88-0) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - 2-Amino-5-nitrophenol is used as a colorant in semipermanent hair dyes and in the manufacture of C.I. Solvent Red 8, an azo dye for synthetic resins, lacquers, and wood stains. 2-Amino-5-nitrophenol was nominated for toxicology and carcinogenesis studies by the National Cancer Institute because of widespread human exposure associated with its use in hair dyes. Toxicology and carcinogenesis studies were conducted by administering 2-amino-5-nitrophenol (98% pure) by gavage in corn oil 5 days per week to groups of F344/N rats and B6C3F1 mice of each sex in 16-day, 13-week, and 2-year studies. In the 2-year studies, male and female rats were given doses of 0, 100, or 200 mg/kg and male and female mice were given doses of 0, 400, or 800 mg/kg. Sixteen-Day and Thirteen-Week Studies: During the 16-day studies, F344/N rats of each sex received 0, 156, 313, 625, 1,250, or 2,500 mg/kg 2-amino-5-nitrophenol by gavage in corn oil vehicle. One of the five males that received 2,500 mg/kg, 1/5 females that received 1,250 mg/kg, and 2/5 females that received 313 mg/kg died before the end of the studies. Final mean body weights of rats that received 1,250 or 2,500 mg/kg were 11% and 30% lower than that of vehicle controls for males and 9% and 13% lower for females. B6C3F1 mice of each sex received doses of 0, 313, 625, 1,250, 2,500, or 5,000 mg/kg 2-amino-5-nitrophenol. Two of five males and 5/5 females that received 5,000 mg/kg, 3/5 males and 3/5 females that received 2,500 mg/kg, 3/5 females that received 1,250 mg/kg, 1/5 females that received 625 mg/kg, and 2/5 male vehicle controls died before the end of the studies. Final mean body weights of chemically exposed mice were not different from those of the vehicle controls. Rats that received 625, 1,250, or 2,500 mg/kg and male mice that received 5,000 mg/kg had loose stools. In 13-week studies, F344/N rats and B6C3F1 mice of both sexes received 0, 100, 200, 400, 800, or 1,600 mg/kg 2-amino-5-nitrophenol by gavage in corn oil. Five of 10 male and 2/10 female rats that received 1,600 mg/kg, 1/10 male and 3/10 female rats that received 800 mg/kg, and 1/10 male rats that received 400 mg/kg died before the end of the studies. Final mean body weights of males that received 400, 800, or 1,600 mg/kg were 10%, 25%, and 43% lower than that of vehicle controls. The final mean body weight of females that received 1,600 mg/kg was 16% lower that of vehicle controls. Four of 10 male and 3/10 female mice that received 1,600 mg/kg died before the end of the 13-week studies. The final mean body weight of male mice that received 1,600 mg/kg was 11% lower than that of vehicle controls; male and female mice that received 1,600 mg/kg appeared lethargic. During the 13-week studies, acute/chronic perivasculitis of vessels of the cecum and colon was observed in rats that received 400, 800, or 1,600 mg/kg and in mice that received 1,600 mg/kg. Body Weight and Survival in the Two-Year Studies: Mean body weights of rats receiving 200 mg/kg were 5%-10% lower than those of vehicle controls after week 33 for males and 4%-5% lower than those of vehicle controls after week 93 for females. Survival of male rats was significantly lower than that of vehicle controls after week 99 for the 100 mg/kg dose group and after week 75 for the 200 mg/kg dose group (final survival: vehicle control, 33/50; 100 mg/kg group, 16/50; 200 mg/kg group, 4/50). Survival of female rats was comparable to that of vehicle controls (30/50; 32/50; 29/50). Loose or poorly formed stools were observed for male rats and occasionally for females that received 200 mg/kg. Mean body weights of mice that received 800 mg/kg were 8%-11% lower than those of vehicle controls between weeks 29 and 74 for males and 8%-13% lower than those of vehicle controls after week 69 for females; mean body weights of mice that received 400 mg/kg were greater than those of vehicle controls after week 69 for males and 5%-9% lower than those of vehicle controls after week 69 for females. Survival of mice that received 800 mg/kg was significantly reduced compared with that of ose of vehicle controls after week 69 for females. Survival of mice that received 800 mg/kg was significantly reduced compared with that of vehicle controls after week 20 for males and week 22 for females and was not considered adequate to evaluate a carcinogenic response (final survival--male: vehicle control, 31/50; 400 mg/kg group, 36/50; 800 mg/kg group, 12/50; female: 37/50; 36/50; 10/50). Nonneoplastic and Neoplastic Effects in the Two-Year Studies: Pigmentation was present at increased incidences in all groups of chemically exposed animals and was characterized by varying amounts of an orange, granular pigment present in the fibrous connective tissue of the lamina propria, in the submucosa, and around vessels in the submucosa of the cecum and colon. Pigmentation of the rectum was observed at increased incidences in male rats that received 100 mg/kg, male and female rats that received 200 mg/kg, and both groups of chemically exposed mice. No pigmentation was found in the intestines of vehicle control rats or mice. Associated with pigmentation was an increased incidence of acute/chronic inflammation in the cecum and colon of all groups chemically exposed rats and mice; this inflammation was similar to that observed in the 13-week studies but was of greater severity. Acute/chronic inflammation was also present in the rectum of male rats that received 100 mg/kg, male and female rats that received 200 mg/kg, and male mice that received 800 mg/kg. The incidence of pancreatic acinar cell adenomas was significantly increased (P≤0.002) in male rats that received 100 mg/kg 2-amino-5-nitrophenol (vehicle control, 1/50; 100 mg/kg, 10/50; 200 mg/kg, 3/49); the increase was considered to be associated with chemical exposure. The reduced survival of male rats that received 200 mg/kg markedly reduced the sensitivity of this group for detecting the presence of neoplasms. The incidences of adenomas or carcinomas (combined) of the preputial or clitoral glands were marginally increased in male or female rats that received 200 mg/kg 2-amino-5-nitrophenol (preputial gland: 3/50; 2/50; 5/50; clitoral gland: 3/50; 3/50; 7/50). Neoplasms found in the intestinal tract of 3/50 male rats that received 100 mg/kg (one leiomyoma of the small intestine, one adenocarcinoma of the jejunum, one leiomyoma of the cecum), 2/50 male rats that received 200 mg/kg (one lipoma and one osteosarcoma of the cecum), and 1/50 female rats that received 200 mg/kg (one leiomyoma of the cecum) were not considered to be the result of chemical exposure. No compound-related neoplasms were found in mice exposed to 2-amino-5-nitrophenol in the 2-year studies. Genetic Toxicology: 2-Amino-5-nitrophenol was mutagenic in Salmonella typhimurium strains TA98, TA100, and TA1537 when tested in a preincubation protocol with and without exogenous metabolic activation, and it exhibited equivocal mutagenic activity in strain TA1535 in the presence of induced liver S9. 2-Amino-5 nitrophenol induced forward mutations in mouse L5178Y lymphoma cells in the absence of metabolic activation; it was not tested with S9. An increase in chromosomal aberrations and sister chromatid exchanges was observed in cultured Chinese hamster ovary (CHO) cells following incubation with 2-amino-5-nitrophenol both in the presence and absence of exogenous metabolic activation. Data Audit: The data, documents, and pathology materials from the 2-year studies of 2-amino-5 nitrophenol were audited at the NTP Archives. The audit findings show that the conduct of the studies is documented adequately and support the data and results given in this Technical Report. Conclusions: Under the conditions of these 2-year gavage studies, there was some evidence of carcinogenic activity for male F344/N rats that received 100 mg/kg 2-amino-5-nitrophenol, as shown by the increased incidence of acinar cell adenomas of the pancreas. Reduced survival of male F344/N rats that received 200 mg/kg decreased the sensitivity of this group for detecting a carcinogenic response. There was no evidence of carcinogenic activity for female rats that received 100 or 200 mg/kg per day. Marginally increased incidences of preputial or clitoral gland adenomas or carcinomas (combined) occurred in male and female F344/N rats administered 200 mg/kg 2-amino-5 nitrophenol. There was no evidence of carcinogenic activity for B6C3F1 mice that received 400 mg/kg 2-amino-5-nitrophenol; reduced survival of B6C3F1 mice that received 800 mg/kg caused this group to be considered inadequate for detecting a carcinogenic response. PMID- 12732903 TI - NTP Toxicology and Carcinogenesis Studies of N-Phenyl-2-naphthylamine (CAS No. 135-88-6) in F344/N Rats and B6C3F1 Mice (Feed Studies). AB - N-Phenyl- 2- naphthylamine, formerly used as a antioxidant in the rubber industry, was selected for toxicology and carcinogenesis studies because at the time of nomination (1976) it had a large annual production and widespread human exposure. Additional reasons for selection included it structural similarity and possible metabolism to the known human urinary bladder carcinogen, 2 naphthylamine. Toxicology and carcinogenesis studies were conducted by feeding diets containing N-phenyl-2-naphthylamine (approximately 98% pure and containing less than 1 ppm 2-naphthylamine) at various concentrations to groups of F344/N rats and B6C3F1 mice of each sex for 14 days, 13 weeks, or 2 years. Fourteen-Day and Thirteen-Week Studies: In 14-day studies, 3/5 male and 4/5 female rats that received 50,000 ppm N-phenyl-2-naphthylamine died before the end of the studies. Final mean body weights of rats that received 12,500 ppm or more were considerably lower (18%-57%) than those of the controls. Arched backs, rough coats, and diarrhea were observed for males that received 12,500 ppm or more and for females that received 25,000 or 50,000 ppm. All mice were alive at the end of the studies, and no compound-related clinical signs of toxicity were observed in mice given feed containing up to 20,000 ppm. In 13-week studies, deaths occurred in 4/10 male and 9/10 female rats that received the highest dose (40,000 ppm) of N-phenyl-2-naphthylamine. Final mean body weights of rats that received 5,000 40,000 ppm were 9%-60% lower than those of the controls. The liver weight to body weight ratios increased with increasing dose, with the ratios for male rats at 10,000 ppm or more and for female rats at 5,000 ppm being greater (P<0.05) than those of controls. A compound-related nephropathy occurred in rats and was characterized by renal tubular epithelial degeneration and hyperplasia. Other effects in rats included hematopoietic hypoplasia or atrophy of the femoral bone marrow, testicular hypospermatogenesis, lymphoid degeneration of the thymus, and lymphoid depletion of the spleen. In mice, 2/10 males and 7/10 females that received 40,000 ppm died before the end of the 13-week studies. The final mean body weights of mice that received 10,000, 20,000, or 40,000 ppm were 9%-32% lower than those of the controls. The liver weight to body weight ratios for mice increased with increasing dose. Those for male mice at 10,000 ppm or more and for female mice at 20,000 ppm or more were greater (P<0.05) than those for the controls. Nephropathywas observed at increased incidences and severity in dosed mice. Because of kidney lesions, liver enlargement, lower weight gain, and increased mortality in the shorter term studies, dietary concentrations of N phenyl-2-naphthylamine selected for the 2-year studies in rats and mice were 0, 2,5000, and 5,000 ppm. Body Weight and Survival in the Two-Year Studies: The mean body weights of dosed rats were lower than those of the controls throughout the studies (12% and 16% lower for dosed males and 15% and 31% lower for dosed females at the end of the studies). The average daily feed consumption for rats was 94%-87% that of the controls for dosed males and 88% that of the controls for dosed females. The estimated average amount of N-phenyl-2-naphthylamine consumed per day was 100 mg/kg and 225 mg/kg for male rats and 120 mg/kg and 260 mg/kg for female rats. The survival of the high dose group of male rats was greater (P<0.05) than that of the controls after week 101 (male: control, 24/50; low dose, 28/50; high dose, 34/50; female: 26/50; 44/50; 38/50). Final mean body weights of high dose male and female mice were lower (male, 9%; female, 23%) than those of the controls. The estimated average daily feed consumption by dosed mice was within 10% that of the controls. The average amount of N-phenyl-2 naphthylamine consumed per day was approximately 500 or 1,000 mg/kg for male mice and 450 or 900 mg/kg for female mice. No significant differences in survival were observed between any groups of mice of either sex (male: control, 33/50; male mice. No significant differences in survival were observed between any groups of mice of either sex (male: control, 33/50; low dose, 36/50; high dose, 28/50; female: 36/50; 30/50; 35/50). Nonneoplastic and Neoplastic Effects in the Two Year Studies: As in the 13-week studies, the kidney was the principal target for toxic effects of N-phenyl-2-naphthylamine. Mineralization of the kidney, necrosis of the renal papilla, and epithelial hyperplasia and calculi of the kidney pelvis were observed at increased incidences in high dose female rats. Hydronephrosis, atrophy, fibrosis, and chronic focal inflammation of the kidney were observed at increased incidences in high dose female rats. Cysts and acute suppurative inflammation of the kidney were observed at increased incidences in dosed male and high dose female rats. No compound-related renal neoplasms were observed in rats. Nuclear enlargement of renal tubular epithelial cells and nephropathy were observed at increased incidences in high dose female mice. Atypical tubular cell hyperplasia occurred in two high dose female mice. A tubular cell adenoma was found in one high dose female mouse, and a tubular cell adenocarcinoma was found in another high dose female mouse. No renal neoplasms were observed in dosed male mice. Neoplasms of several organs occurred in rats with negative trends and/or at significantly lower incidences in high dose groups. These included thyroid gland C-cell neoplasms in males and females and mammary gland fibroadenomas, pituitary gland adenomas, and mononuclear cell leukemia in females. The lack of carcinogenicity in rats may be related to an inability to metabolize this compound to the known animal and human carcinogen 2-napththylamine. Genetic Toxicity: N-Phenyl-2-naphthylamine was not mutagenic in the Salmonella typhimurium/microsome assay with strains TA97, TA98, TA100, or TA1535 with or without induced hamster or rat liver S9. The chemical did not induce chromosomal aberrations in cultured Chinese hamster ovary (CHO) cells with or without metabolic activation. No increase in sister chromatid exchanges (SCEs) was observed in the absence of metabolic activation; in the presence of rat liver S9, the SCE results were judged to be equivocal. Data Audit: The data, documents, and pathology materials from the 2-year studies of N-phenyl-2-naphthylamine were audited at the NTP Archives. The audit findings show thatthe conduct of the studies is documented adequately and support the data and results given in this Technical Report. Conclusions: Under the conditions of these 2-year feed studies, there was no evidence of carcinogenic activity for male or female F344/N rats fed diets containing 2,500 or 5,000 ppm N-phenyl-2-naphthylamine. Decreased incidences of several neoplasms were observed in dosed rats: thyroid gland C-cell neoplasms in males and females and mononuclear cell leukemia, pituitary gland adenomas, and mammary gland fibroadenomas in females. There was no evidence of carcinogenic activity for male B6C3F1 mice fed diets containing 2,500 or 5,000 ppm N-phenyl-2-naphthylamine. There was equivocal evidence of carcinogenic activity of N-phenyl-2-naphthylamine for female B6C3F1 mice as indicated by the occurrence of two rare kidney neoplasms. Chemical-related nonneoplastic lesions (nephropathy, karyomegaly, and hyperplasia) occurred in the kidney of rats and mice. Synonyms: N-(2-naphthyl)aniline; 2-naphthylphenylamine; b naphthylphenylamine; 2-phenylaminonaphthalene; phenyl-b-naphthylamine; N-phenyl-b naphthylamine Trade Names: Aceto PBN; Agerite Powder: Antioxidant 116; Neosone D; Neozon D; Nilox PBNA; Nonox D; PBNA; Stabilizator AR PMID- 12732904 TI - NTP Toxicology and Carcinogenesis Studies of 2-Mercaptobenzothiazole (CAS No. 149 30-4) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - Toxicology and carcinogenesis studies of technical-grade 2-mercaptobenzothiazole (96%-97% pure), a rubber accelerant and preservative, were conducted by administering the chemical by gavage in a corn oil vehicle to groups of F344/N rats and B6C3F1 mice of each sex for 16 days, 13 weeks, or 2 years. 2 Mercaptobenzothiazole was nominated for study by the National Institute of Environmental Health Sciences and the National Institute for Occupational Safety and Health. Sixteen-Day and Thirteen-Week Studies: In 16-day studies, mean body weight gains of rats receiving 2,500 mg/kg were 6-7 g lower than those of vehicle controls; 4/5 male and 5/5 female mice dosed with 3,000 mg/kg and 4/5 female mice dosed with 1,500 mg/kg died; lethargy and prostration occurred in most of these animals after gavage. Based on these results, doses were selected for both species in the 13-week studies were 0, 94 (mice only), 188, 375, 750, and 1,500 mg/kg. In the 13-week studies, no chemical-related deaths occurred in rats, but body weight gains in males dosed with 1,500 mg/kg and in females dosed with 750 or 1,500 mg/kg were lower than those in the vehicle control groups. Hepatomegaly occurred at the two highest doses in males and at all doses in females; however, no microscopic pathologic changes were noted in any tissue. More than half the mice dosed with 1,500 mg/kg died, but no compound-related body weight changes occurred. Clinical signs in mice were dose related and included lethargy in animals dosed with 375 mg/kg and lacrimation, salivation, and clonic seizure in some dosed with 750 or 1,500 mg/kg. No association between these clinical signs of toxicity and gross or microscopic pathologic effects were observed. Doses selected for the 2-year studies were 0, 375, and 750 mg/kg for male rats and for mice of each sex and 0, 188, or 375 mg/kg for female rats. Body weight and Survival in the Two-Year Studies: Fifty animals of each species and sex were administered 2-mercaptobenzothiazole in corn oil by gavage 5 days per week for 103 weeks. Administration of 2-mercaptobenzothiazole resulted in decreased survival in dosed male rats (vehicle control, 42/50; low dose, 22/50; high dose, 20/50) and in the high dose group of female mice (37/50; 39/50; 22/50) but not in female rats (28/50; 31/50; 25/50) or in male mice (38/50; 33/50; 30/50). No effect on body weight gain in dosed rats was observed; in dosed mice, minor reductions occurred between weeks 3 and 64, withrecovery thereafter. Postgavage lethargy and prostration occurred frequently in dosed rats and mice. Nonneoplastic and Neoplastic Effects in the Two-Year Studies: The severity of nephropathy was increased in dosed male rats. Ulcers and inflammation of the forestomach were prevalent in dosed rats, as were increased incidences of epithelial hyperplasia and hyperkeratosis in male rats, but no neoplasms of the forestomach were observed. There were no increases of nonneoplastic lesions in mice which were considered to be compound related. The incidences of a variety of tumors were increased in rats dosed with 2-mercaptobenzothiazole; some of the increased incidences were not dose related. In low dose male rats, increased incidences (P<0.01) were observed for mononuclear cell leukemia (7/50; 16/50; 3/50) and pancreatic acinar cell adenomas (2/50; 13/50; 6/49). Increased tumor incidences with dose-related trends (P<0.05) included pituitary gland adenomas in females (15/49; 24/50; 25/50), preputial gland adenomas or carcinomas (combined) in males (1/50; 6/50; 5/50), adrenal gland pheochromocytomas or malignant pheochromocytomas (combined) in males (18/50; 27/50; 24/49), and pheochromocytomas in females (1/50; 5/50; 6/50). These tumors were observed at significantly greater incidences (P40 units) prior to transplantation as being associated with higher mortality in renal transplant recipients (risk ratio (RR): 3.1, confidence interval (CI): 1.1-9.2; P=0.03). These data suggest that serum ferritin levels above 1100 ng/ml due to multiple blood transfusions causing iron overload is a relevant factor that increases mortality. PMID- 12732930 TI - Transfection of normal primary human skeletal myoblasts with p21 and p57 antisense oligonucleotides to improve their proliferation: a first step towards an alternative molecular therapy approach of Duchenne muscular dystrophy. AB - Duchenne muscular dystrophy (DMD), caused by the absence of dystrophin, is associated with decreased muscle cell proliferation. An increased p21 mRNA level in DMD patients may be involved in the process. In this context we are interested to improve the proliferation of primary human skeletal muscle cells (SkMC) by a reduction in the cell cycle proteins p21 and p57 using the appropriate antisense oligonucleotides (ASO). Therefore a transfection procedure needs to be optimized in which the oligonucleotide enters the SkMC with a minimal loss of cell vitality and high efficiency. Three different formulations, Effectene, DAC40, and SuperFect, were compared. Proliferation was analyzed comparing cells transfected with p21 and/or p57 ASO vs. cells transfected with scrambled ASO using a bromodeoxyuridine assay. Under optimal conditions (a mixture of 0.25 microg ASO, 5 microl Effectene, 0.8 microl enhancer) SkMC transfected with p21 ASO reveal an average increase in cell proliferation of 32.5+/-11% after 24 h. p57 ASO shows the same effect, but concomitant transfection of p21 and p57 does not enhance it. A cell vitality of 78+/-14% after 24 h was determined by the MTT test. SkMC transfected with DAC40 reveal a maximal increase in proliferation of 38+/-7% after 48 h and show a vitality of 65+/-8%. In contrast to both these formulations, SuperFect was found to be highly toxic for SkMC, with more than 70% dead cells after 24 h. The increase in proliferation, the functional biological effect of p21 ASO, is well correlated with a decrease in p21 detected by western blot analysis of 31.6% for Effectene. Transfection efficiency was measured directly by FACS analysis using FITC-labeled ASO and data showing ASO internalization in 75.8+/-11.2% of the cell population for Effectene and 74.4+/ 6.6% cells for DAC40. Taken together transient transfection of p21 or p57 ASO into primary human SkMC using Effectene significantly improves their proliferation compared to transfection with scrambled ASO without a major loss of cell vitality. This represents a basis for the transfer of this technique to dystrophin-deficient SkMC cultures and the introduction of the short interference RNA technique which might enhance the effect on cell proliferation. PMID- 12732931 TI - Amniotic fluid embolism with involvement of the brain, lungs, adrenal glands, and heart. AB - The case of a healthy 31-year-old woman in the 40th week of second pregnancy is presented. During preparation for an emergency caesarean section, she developed an amniotic fluid embolism (AFE) with unusual and unique features. The acute onset of disease with cardiorespiratory failure with hypotension, tachycardia, cyanosis, respiratory disturbances and loss of consciousness, suggested at first a pulmonary thromboembolism, but the appearance of convulsions led to the diagnosis of AFE. The patient died after 5 days due to an untreatable brain edema. At autopsy, AFE with the usually associated disseminated intravascular coagulation was found in the lungs, brain, left adrenal gland, kidneys, liver and heart. Eosinophilic inflammatory infiltrates were found in the lungs, hepatic portal fields and especially in the heart, suggesting a specific hypersensitivity reaction to fetal antigens. Moreover, intravascular accumulation of macrophages in the lungs also favored a non-specific immune reaction to amniotic fluid constituents. PMID- 12732932 TI - Post-tonsillectomy haemorrhage: a 7-year retrospective study. AB - Amongst fears about iatrogenic transmission of new variant Creutzfeldt-Jakob disease (vCJD), last year the Department of Health of the U.K. advised that all adenotonsillectomies be performed using disposable instruments. Following implementation of these guidelines, many consultants reported an increase in post tonsillectomy haemorrhage in association with the use of disposable instruments. We have undertaken a retrospective study of tonsillectomy and have confirmed a significant increase in the incidence of return to the theatre for arrest of postoperative haemorrhage (5.32% vs. 2.18%) during the period when disposable instruments were used. PMID- 12732933 TI - Anaesthesia in endoscopic sinus surgery. AB - The aim of this study was to evaluate the methods of anaesthesia used in 1,460 patients (677 females and 783 males), aged 4 to 79 years, who were consecutively operated on with a functional endonasal endoscopic approach (FESS) or by an endoscopic endonasal approach (ESS) by one surgeon (AD) in the period from 1987 2001. The included patients presented a variety of disorders within the nose and sinuses. Many of the patients had sinonasal polypoid disease, which had gradually expanded over years and was causing pressure to neighbouring structures. Several had undergone previous multiple surgical procedures altering the original anatomy. However, the majority of the patients had been treated surgically for acute recurrent and/or chronic infections, others for sinogenic headache, benign tumours/mucoceles and smell disorders. The intention of looking into the anaesthetic procedures being used was to clarify the needs for a differentiated anaesthetic approach to different surgical tasks. It is of crucial importance for an optimal surgical result that both the patient and surgeon feel absolutely comfortable during the operation. Local anaesthesia can be used in minor surgery on selected patients and is definitely suitable even for residents in training. General anaesthesia is preferred in most cases by both patients and surgeons. The conclusion of this paper is that total intravenous anaesthesia (TIVA) in addition to oxygen-enriched air through a laryngeal mask airway (LMA) should be the anaesthetic procedure of choice, at least in our experience. PMID- 12732934 TI - Endoluminal stenting for tracheal stenosis. AB - This prospective clinical study evaluates the role of endoluminal stents in the maintenance of a tracheal lumen after management of stenosis. Tracheal stenosis may be due to a variety of causes, and it is a significant health problem. Most patients either remain tracheostomized for prolonged periods of time or undergo several major surgical interventions. The most common sequel is restenosis at the site of repair either after augmentation or resection anastomosis. In this study, 16 patients with different pathologies causing tracheal stenosis or collapse are presented. All patients had the tracheal lumen re-established either endoscopically or by an open procedure. The stent was then placed to support the site of repair and prevent restenosis. A patent well-epithelialized lumen was achieved in 11 cases with a follow-up of 3-24 months. Complications were encountered in five cases: three misplaced stents, one tracheal erosion with a Dumon stent and one granulomatous obstruction. The technique, complications and follow-up of these patients are described with recommendations for the management of such patients. PMID- 12732935 TI - Clustering of neuronal inclusions in "dementia with neurofilament inclusions". AB - Dementia with neurofilament inclusions (DNI) is a new disorder characterized clinically by early-onset dementia and histologically by the presence of intraneural inclusions immunopositive for neurofilament antigens but lacking tau and alpha-synuclein reactivity. We studied the clustering patterns of the neurofilament inclusions (NI) in regions of the temporal lobe in three cases of DNI to determine whether they have the same spatial patterns as inclusions in the tauopathies and alpha-synucleinopathies. The NI exhibited a clustered distribution (mean size of clusters 400 microm, range 50-800 microm, SD 687.8) in 24/28 of the areas studied. In 22 of these areas, the clusters exhibited a regular distribution along the tissue parallel to the pia mater or alveus. In 3 cortical areas, there was evidence of a more complex pattern in which the NI clusters were aggregated into larger superclusters. In 6 cortical areas, the size of the clusters approximated to those of the cells of origin of the cortico cortical pathways but in the remaining areas cluster size was smaller than 400 microm. Despite the unique molecular profile of the NI, their spatial patterns are similar to those shown by filamentous neuronal inclusions in the tauopathies and alpha-synucleinopathies. PMID- 12732936 TI - Distribution of astrocytic plaques in the corticobasal degeneration brain and comparison with tuft-shaped astrocytes in the progressive supranuclear palsy brain. AB - Corticobasal degeneration (CBD) and progressive supranuclear palsy (PSP) have some clinical and pathological features in common. Each, however, has been shown to have specific astrocytic inclusions. CBD is characterized by astrocytic plaques, and PSP is characterized by tuft-shaped astrocytes. To clarify differences between these inclusions, we investigated intracerebral distribution of astrocytic plaques and tuft-shaped astrocytes in autopsied brains of patients with either CBD or PSP. Specimens from ten patients with CBD and five patients with PSP were stained by the Gallyas-Braak method. Densities of the astrocytic plaques and tuft-shaped astrocytes were determined for 11 cerebral cortical regions, 6 subcortical nuclei, 5 brain stem regions, the cerebellar cortex and the dentate nucleus. Astrocytic plaques were most abundant in the prefrontal and premotor areas in the cerebral cortex of CBD brains, whereas tuft-shaped astrocytes were most prominent in the precentral gyrus and premotor area of PSP brains. Many astrocytic plaques were observed in the caudate nucleus of CBD brains, whereas tuft-shaped astrocytes were abundant in both the caudate and putamen and moderate in number in the globus pallidus, subthalamic nucleus and thalamus in PSP brains. Very slight accumulation of astrocytic plaques was seen in the brain stem of CBD brains, whereas numerous tuft-shaped astrocytes were found in the red nucleus and superior colliculus of PSP brains. Distribution of the astrocytic plaques and tuft-shaped astrocytes differed greatly. Thus, CBD and PSP can be considered different entities. PMID- 12732937 TI - The effect of hypoxia on permeability and bacterial translocation in Caco-2 adult and I-407 fetal enterocyte cell culture models. AB - Hypoxia has been implicated in the breakdown of the intestinal epithelial barrier in animals, leading to bacterial translocation (BT); however, the mechanism of this hypoxic insult is unknown. To determine the effects of hypoxic injury in vitro on epithelial membrane integrity, transepithelial electrical resistance (TEER), mannitol permeability (Ma-Pm), and BT were measured in both an adult (Caco-2) and fetal (I-407) intestinal epithelial cell culture model. Caco-2 adult and I-407 fetal epithelial cell monolayers were treated with or without bacteria (1 x 10(7) Escherichia coli. C-25), and then incubated under either normoxic (5% CO(2) in room air) or hypoxic (5% CO(2) and 95% N(2)) conditions at 37 degrees C for 6 h. Hypoxia caused a 10% increase in Ma-Pm in the I-407 fetal cell model independent of the bacterial challenge. In contrast, a bacterial challenge in the Caco-2 adult model caused a 485% increase in Ma-Pm independent of hypoxia. Neither hypoxia, nor C-25 bacteria, for 6 h caused BT in either cell culture model. In the adult cell culture model, bacteria appear to mediate changes in epithelial barrier function, with hypoxia having no effect. On the other hand, hypoxia is the major factor in the loss of epithelial barrier function in fetal epithelium, but has no effect on adult epithelium. The data suggest that the breakdown of barrier function caused by a hypoxic insult is the primary stimulus for subsequent BT in neonates. PMID- 12732938 TI - Interleukin-6 changes tight junction permeability and intracellular phospholipid content in a human enterocyte cell culture model. AB - Proinflammatory cytokines and secretory phospholipase A(2) (sPLA(2)) are elevated in patients with inflammatory bowel disease (IBD). We previously reported that the proinflammatory cytokine IL-6 increased the expression of sPLA(2) (a hydrolyzer of phosphatidylcholine) and decreased membrane integrity in an intestinal epithelial cell culture model. To determine the physiological effects of the IL-6 mediated increase in sPLA(2) on decreased epithelial layer integrity, we investigated alterations of intracellular/secretory phospholipid (PL) composition in a cell culture model. In addition, since other PLs may also mediate epithelial membrane activity, we investigated the effect of IL-6 on PL activity in a Caco-2 enterocyte culture model. Caco-2 cells were incubated for 72 h with IL-6 or media alone (control). Both media and cell lysate were analyzed for PL composition using thin-layer chromatography. The PL composition in the media did not show any differences between the two groups ( p>0.1). Total intracellular PL contents were also unchanged; however, IL-6 led to significant changes in PL composition including an increase in phosphatidylethanolamine (PE) and sphingomyelin (SM) and a decrease in phosphatidylcholine (PC) and lysophosphatidylcholine (LPC) ( p<0.05). Both PE and SM are known as inflammatory signaling factors involved in human IBD. Our study suggests that the decreased membrane integrity seen with IL-6 application may occur via intracellular PL alterations, rather than through the direct effects of sPLA(2). PMID- 12732940 TI - Brain tumors in children under 1 year of age: emphasis on the relationship of prognostic factors. AB - OBJECTS: Primary brain tumors in infants under 12 months of age have a different prognosis from older children. MATERIAL: A retrospective analysis was done in all patients less than 12 months old with primary brain tumors. RESULTS: Out of 1682 children with primary brain tumors, 61 (3.6%) were infants under 12 months old. The mean age at diagnosis was 181.6 days (SD 128) with a range of 1 to 364 days. There were 37 males (60.6%). The most common tumor was astrocytoma ( n=22) (36%). Supratentorial tumors were present in 63.9% but this was not related to survival ( p=0.1095). Complete surgical resection ( n=14) was favorable for survival ( p=0.039). Intracranial hypertension at diagnosis did not influence survival ( p=0.89). The overall survival rate was 32%, mean 42.08 (SD 7.38). A total of 24 patients are alive and without evidence of disease. CONCLUSION: Complete surgical resection was necessary for a favorable prognosis, and the long-term effects are a valid problem. PMID- 12732941 TI - Brain tumors in children under 1 year of age: emphasis on the relationship of prognostic factors. PMID- 12732939 TI - Stereotactically-guided biopsies of brainstem tumors. AB - INTRODUCTION: More than 10 years ago, the goal of our work had been to obtain a tissue sample of infiltrating lesions of the brainstem that had been diagnosed using computerized axial tomography (CAT). At that time, biopsies were believed to be indispensable when starting treatment of tumors. With time our objectives changed. Biopsies remained necessary, since until 1 year before the writing of this article we had not had the benefits of magnetic resonance imaging (MRI) at our Hospital. We also decided that carrying out sound statistics, confirmed by biopsies, was in itself a good procedure, especially in a country in which, to date, no serial studies of brainstem tumors had been undertaken. METHODS: We analyzed all of the patients diagnosed with posterior fossa tumors between March 1989 and March 2002 at the Hospital Infantil de Mexico Federico Gomez (HIM). A preoperative TAC of the cranium was performed on every patient. Stereotactically guided biopsies during tomography allowed precise control of penetration. Material obtained was sent to the Department of Pathology for analysis. RESULTS: Fifty patients were diagnosed with infiltrating tumors of the brainstem: 30 cases of low-grade astrocytomas, 13 cases of high-grade astrocytomas, 2 cases of primitive neuroectodermic tumors, 2 cases of rhabdoid tumors, 1 case of ependymoma, and 2 patients had non-specified tumors. The most frequent symptoms and signs were ataxia and disturbances of the cranial nerves. There was no mortality caused by penetration, and follow-up studies of more than 5 years were carried out. DISCUSSION: The results from our series were similar to those in the literature. In our case, follow-up studies were undertaken for longer periods. In the first section of our work, we suggest the need for stereotactic biopsies in order to arrive at a precise diagnosis in environments in which MRI may be unavailable. CONCLUSION: At present, presumptive diagnosis of infiltrating brainstem lesions may be adequately undertaken with imaging methods, such as MRI. However, we believe that a stereotactically-guided biopsy provides an accurate method for diagnosing lesions of the brainstem. In our case, this procedure has been carried out entirely in the tomography room, without any complications of disease or mortality. PMID- 12732943 TI - Identification of the gene for beta-fructofuranosidase of Bifidobacterium lactis DSM10140(T) and characterization of the enzyme expressed in Escherichia coli. AB - Bifidobacterium lactis is a moderately oxygen-tolerant, saccharolytic bacterium often used in combination with fructooligosaccharides (FOS) as a probiotic supplement in diverse dairy products. This is the first report describing the gene structure and enzymatic properties of a beta-fructofuranosidase [EC 3.2.1.26] from Bifidobacteria. BfrA was identified in Bifidobacterium lactis DSM 10140(T) and heterologously expressed in Escherichia coli. The G+C content was identical with the G+C content as determined for the total genomic DNA (61.9 mol %). The gene codes for a 532-aa residue polypeptide of 59.4 kDa. Surprisingly, the deduced aa sequence revealed only minor similarity to other fructofuranosidases (18% to E. coli cscA). The enzyme was purified to homogeneity after incorporation of a C-terminal 6 x HIS affinity tag. It hydrolased sucrose, 1-kestose, Raftilose, Actilight, inulin, and raffinose (100%, 91%, 84%, 80%, 37%, 4%). Fructose moieties were released in an exo-type fashion. Substrates with alpha-glycosidic linkages or residues other than fructose were not attacked. The kinetic parameters K(m) and V(max) for sucrose hydrolysis were 10.3 m M and 0.031 microM/min (pH 7.6; 37 degrees C). The activity was abolished by Zn(2+) (1 m M) and significantly inhibited by Fe(2+) and Ni(2+) (10 m M). The enzyme showed its maximal activity at 40 degrees C. PMID- 12732942 TI - Morphological and functional investigations of neuroendocrine tumors of the pancreas. AB - Neuroendocrine tumors of the pancreas are rare neoplasms arising predominantly from the pancreatic islets of Langerhans and are thus known as islet cell tumors. More than the half of all neuroendocrine tumors are called functioning islet cell tumors because they secrete one or more biologically active peptides that may produce clinical symptoms. Clinical diagnosis of non-functioning, i.e., biologically inactive, tumors is often delayed and patients tend to present with advanced tumors (size greater than 5 cm) that are easily localized by using conventional imaging modalities. On the other hand, symptoms of functioning islet cell tumors usually appear early in the clinical course, rendering the preoperative localization of these small hormone-producing tumors (size less than 2 cm) difficult with non-invasive methods. Since functioning islet cell tumors can often be cured by surgical resection, invasive procedures are warranted when necessary for localization diagnosis. Failure to search for, detect, and resect these small tumors will invariably result in persistent symptoms. Regarding the unsatisfactory results of morphological imaging methods, functional studies, especially arterial stimulation with hepatic venous samplings, may provide a preoperative regionalization of the pancreatic adenoma, regardless of its size. PMID- 12732944 TI - Modeling lipid accumulation and degradation in Yarrowia lipolytica cultivated on industrial fats. AB - A modeling approach was used to quantify the kinetic behavior of a Yarrowia lipolytica strain capable of producing significant lipid amounts when cultivated on industrial fats. Biomass and cellular lipid evolution were successfully simulated, while the optimized parameter values were similar to those experimentally measured. The maximum specific formation rate of fat-free biomass seemed unaffected by the substrate fatty acid composition. On the contrary, the maximum concentration of lipid accumulated inside the yeast cell, as well as the maximum specific accumulation rate of cellular lipids, was favored in high stearic acid content media. The microorganism presented the tendency to degrade its accumulated lipids, although remarkable substrate fat amounts remained unconsummated in the culture medium. This degradation slowly occurred in the yeast cell as the specific rate of the intracellular carbon pool (storage lipid consumption) was significantly lower compared with that of the extracellular carbon pool (substrate fat). However, the fat-free biomass yield on storage lipids (g of fat-free biomass formed per g of storage lipids consumed) was higher than the one on the substrate (g of fat-free biomass formed per g of medium fat consumed). PMID- 12732945 TI - Structure and Function of sanV: a gene involved in nikkomycin biosynthesis of Streptomyces ansochromogenes. AB - A 6.3-kb BamHI- BglII DNA fragment was cloned from cos20 by using chromosome walking strategy. It was partially sequenced with the result that there is a possible ORF of 1272 nucleotides. The ORF designated sanV was deposited in GenBank under accession no. AF469955. Database search indicated that the deduced protein of sanV shows 28% identity and 44% similarity over 405 amino acid residues to the large component (E) of glutamate mutases from Clostridium cochlearium. Gene disruption was performed to study the function of sanV. It was found that sanV disruptants exhibited much poorer inhibition to the test strain than the wild-type S. ansochromogenes 7100. Furthermore, HPLC analysis indicated that the sanV disruptants almost did not produce nikkomycins X and Z, whereas they accumulated new nikkomycins O(x) and O(z), which revealed that sanV is an important gene involved in the biosynthesis of the peptidyl moiety of nikkomycins. PMID- 12732946 TI - Neurospora crassa FKS protein binds to the (1,3)beta-glucan synthase substrate, UDP-glucose. AB - The essential fungal cell-wall polymer (1,3)beta-glucan is synthesized by the enzyme (1,3)beta-glucan synthase. This enzyme, which is the target of the echinocandin and pneumocandin families of fungicidal antibiotics, is a complex composed of at least two proteins, Rho1p and Fks1p. Homologs of the yeast FKS1 gene have been discovered in numerous fungi, and existing evidence points to, but has not yet proved, Fks1p being the catalytic subunit of (1,3)beta-glucan synthase. We have purified (1,3)beta-glucan synthase from Neurospora crassa approximately 400-fold enrichment and labeled the substrate-binding protein by using a UDP-glucose analog, 5-azido-[beta-(32)P]-UDP-glucose. UDP-glucose-binding proteins were photo-crosslinked to the substrate analog and identified from SDS PAGE gels by Quadrupole time-of-flight mass spectrometry by sequencing the tryptic peptides. Two plasma membrane proteins were labeled FKS and H(+)-ATPase. These results suggest that FKS appears to be the substrate-binding subunit of (1,3)beta-glucan synthase. PMID- 12732947 TI - Identification and characterization of a shuttle plasmid with antibiotic resistance gene from Staphylococcus aureus. AB - While studying antibiotic-resistant plasmids from multi-drug-resistant nosocomial Staphylococcus aureus strains, we isolated a small (2.889 kb) chloramphenicol resistant (Cm(r)) plasmid, which was designated as pMC524/MBM. The molecular size of pMC524/MBM was close to that of pC194 (2.910 kb), a well-known Cm(r) staphylococcal plasmid. Unlike pC194, this plasmid can replicate and express itself efficiently and stably in Escherichia coli. However, Cm is needed for stable maintenance of pMC524/MBM in different hosts. In this study, the nucleotide sequences of these two plasmids were compared after sequencing of pMC524/MBM [EMBL Accession No. AJ312056 SAU312056]. Although these two plasmids have striking nucleotide sequence homology, the Plus Origin, Minus Origin, the replication protein (Rep), and the chloramphenicol acetyl transferase (Cat) have considerable variations. Possibly, these changes have modulated pMC524/MBM into an efficient shuttle-plasmid. PMID- 12732948 TI - Superoxide dismutase is a virulence factor produced by the coral bleaching pathogen Vibrio shiloi. AB - Coral bleaching is a disease that threatens coral reefs throughout the world. The disease is correlated with higher-than-normal seawater temperatures. Data have been reported showing that bleaching of the coral Oculina patagonica during the summer in the Mediterranean Sea is the result of an infection with Vibrio shiloi. The summer temperatures induce the expression of virulence factors in the pathogen. We report here that V. shiloi produces an extracellular superoxide dismutase (SOD) at 30 degrees C, but not at 16 degrees C. An SOD(-) mutant was avirulent. The mutant adhered to corals, penetrated into coral cells, multiplied intracellularly for a short time, and then died. These data support the hypothesis that SOD protects the intracellular V. shiloi from oxidative stress caused by the high concentration of oxygen produced by intracellular zooxanthellae photosynthesis. PMID- 12732950 TI - Involvement of glycine and aspartate residues in the binding capacity of FAD in the NADH dehydrogenase from an alkaliphilic Bacillus. AB - There is a region exhibiting a similarity of amino acid sequence near the carboxyl-terminal segment of each FAD-containing oxidoreductase. In this region, four amino acid residues-Thr, Ala, Gly, and Asp-are highly conserved. To determine the involvement of the four amino acid residues (Thr-469, Ala-476, Gly 478, and Asp-479) in the activity of NADH dehydrogenase of an alkaliphilic Bacillus, mutations of these amino acid residues were conducted. In spite of high conservation, mutations of Thr-469 and Ala-476 to Ala and Ser, respectively, did not lead to a critical loss of enzyme activity. However, mutations of Gly-478 and Asp-479 to Ala caused a complete loss of the activity, which appears to result from the loss of binding capacity of FAD. PMID- 12732949 TI - Altered kinetic properties of tyrosine-183 to cysteine mutation in glutamine synthetase of anabaena variabilis strain SA1 is responsible for excretion of ammonium ion produced by nitrogenase. AB - A L-methionine- D, L-sulfoximine-resistant mutant of the cyanobacterium Anabaena variabilis, strain SA1, excreted the ammonium ion generated from N(2) reduction. In order to determine the biochemical basis for the NH(4)(+)-excretion phenotype, glutamine synthetase (GS) was purified from both the parent strain SA0 and from the mutant. GS from strain SA0 (SA0-GS) had a pH optimum of 7.5, while the pH optimum for GS from strain SA1 (SA1-GS) was 6.8. SA1-GS required Mn(+2) for optimum activity, while SA0-GS was Mg(+2) dependent. SA0-GS had the following apparent K(m) values at pH 7.5: glutamate, 1.7 m M; NH(4)(+), 0.015 m M; ATP, 0.13 m M. The apparent K(m) for substrates was significantly higher for SA1-GS at its optimum pH (glutamate, 9.2 m M; NH(4)(+), 12.4 m M; ATP, 0.17 m M). The amino acids alanine, aspartate, cystine, glycine, and serine inhibited SA1-GS less severely than the SA0-GS. The nucleotide sequences of glnA (encoding glutamine synthetase) from strains SA0 and SA1 were identical except for a single nucleotide substitution that resulted in a Y183C mutation in SA1-GS. The kinetic properties of SA1-GS isolated from E. coli or Klebsiella oxytoca glnA mutants carrying the A. variabilis SA1 glnA gene were also similar to SA1-GS isolated from A. variabilis strain SA1. These results show that the NH(4)(+)-excretion phenotype of A. variabilis strain SA1 is a direct consequence of structural changes in SA1-GS induced by the Y183C mutation, which elevated the K(m) values for NH(4)(+) and glutamate, and thus limited the assimilation of NH(4)(+) generated by N(2) reduction. These properties and the altered divalent cation mediated stability of A. variabilis SA1-GS demonstrate the importance of Y183 for NH(4)(+) binding and metal ion coordination. PMID- 12732951 TI - Binding of extracellular matrix molecules by enterococci. AB - The bacterial surfaces of enterococci are not uniform. This fact is confirmed by several studies and by our results when great differences between individual strains with regard to their cell surface hydrophobicity, binding of eight ECM (extracellular matrix) molecules immobilized on latex beads and four selected ECM molecules in microtiter plates were observed. The strains expressing high binding of ECM molecules (e.g., HJ 18, HJ 23, HJ 24, HJ 26, HJ 28, HJ 36, etc.) were found among Enterococcus faecalis and E. faecium by PAA (particle agglutination assay). On the other hand, weak ECM binders (e.g., HJ 21, HJ 32, HJ 34, HJ 38, HJ 39, HJ 42, HJ 43) were also found. A direct correlation was found between porcine mucin and fetuin binding ability of eight selected strains tested in microtiter plates and by PAA. Moreover, the influence of tunicamycin treatment was different because significant (P < 0.001) blocking effect of tunicamycin was observed with two selected strains (HJ 26 and HJ 36), whereas two strains (HJ 18 and HJ 22) were not significantly affected in their fetuin binding. The treatment of six enterococcal strains with proteolytic enzymes, pronase P, and trypsin, and with sodium metaperiodate also significantly (P < 0.001) decreased their fetuin binding. This suggests that both protein and carbohydrate moieties are involved in the binding of immobilized fetuin. However, the influence of these chemicals on the fetuin binding by individual strains was different. PMID- 12732952 TI - Detection of Actinobacillus pleuropneumoniae by PCR on field strains from healthy and diseased pigs. AB - We investigated whether primers able to specifically amplify a 0.7-kb DNA fragment from the conserved cpx genes could be applied to analyze A. pleuropneumoniae field isolates. The specific cpx primers were tested on 120 strains of A. pleuropneumoniae and other NAD-dependent field isolates from healthy and diseased animals to analyze A. pleuropneumoniae isolates from pigs in Brazil. We found that PCR and hybridization were able to discriminate between isolates of A. pleuropneumoniae and other bacteria. The 0.7-kb cpx DNA fragments were amplified from all 63 A. pleuropneumoniae isolates from herds with clinical symptoms and were isolated from lesions of acute cases of swine pleuropneumonia, both serotypable and nonserotypable. The PCR was also applied to 57 field isolates obtained from animals of apparently healthy herds, and the amplified cpx product was present in four serotypable and only two out of eleven A. pleuropneumoniae nonserotypable isolates. All nonserotypable A. pleuropneumoniae isolates revealed the apxA amplification pattern compatible with previously known serotypes. Some nonserotypable isolates might represent a population of isolates that originally were serotypable but lost the ability to react with serotype specific antisera or might belong to novel serotypes. The PCR method applied is highly sensitive for serotypable A. pleuropneumoniae strains and for nonserotypable strains isolated from acute cases of swine pleuropneumoniae in Brazil. PMID- 12732953 TI - A novel, highly viscous polysaccharide excreted by an alteromonas isolated from a deep-sea hydrothermal vent shrimp. AB - A deep-sea, mesophilic, aerobic, and heterotrophic microorganism, able to produce an extracellular polysaccharide, was isolated from a shrimp collected near an active hydrothermal vent of the Mid-Atlantic Ridge. On the basis of phenotypic and phylogenetic analyses and DNA/DNA relatedness, this strain could be assigned to the species Alteromonas macleodii as a variant of the fijiensis subspecies. It was selected for its ability to exhibit a swarming mucoid phenotype on specific media. The bacterium secreted, under laboratory conditions, an extremely viscous exopolysaccharide consisting of glucose, galactose as neutral sugars, and glucuronic, galacturonic acids as uronic acids, along with pyruvate and acetate as main substituents. PMID- 12732954 TI - A 50S ribosomal subunit precursor particle is a substrate for the ErmC methyltransferase in Staphylococcus aureus cells. AB - Macrolide antibiotics like erythromycin can induce the synthesis of a specific 23S rRNA methyltransferase which confers resistance to cells containing the erm gene. Erythromycin inhibits both protein synthesis and the formation of 50S subunits in bacterial cells. We have tested the idea that the 50S precursor particle that accumulates in antibiotic-treated Staphylococcus aureus cells is a substrate for the methyltransferase enzyme. Pulse-chase labeling studies were conducted to examine the rates of ribosomal subunit formation in control and erythromycin-induced cells. Erythromycin binding to 50S subunits was examined under the same conditions. The rate of 50S subunit formation was reduced for up to 30 min after antibiotic addition, and erythromycin binding was substantial at this time. A nuclease protection assay was used to examine the methylation of adenine 2085 in 23S rRNA after induction. A methyl-labeled protected RNA sequence was found to appear in cells 30 min after induction. This protected sequence was found in both 50S subunits and in a subunit precursor particle sedimenting at about 30S in sucrose gradients. 23S rRNA isolated from 50S subunits of cells could be labeled by a ribosome-associated methlytransferase activity, with (3)H-S adenosylmethionine as a substrate. 50S subunits were not a substrate for the enzyme, but the 30S gradient region from erythromycin-treated cells contained a substrate for this activity. These findings are consistent with a model that suggests that antibiotic inhibition of 50S formation leads to the accumulation of a precursor whose 23S rRNA becomes methylated by the induced enzyme. The methylated rRNA will preclude erythromycin binding; thus, assembly of the particle and translation become insensitive to the inhibitory effects of the drug. PMID- 12732955 TI - The role of the sigB gene in the general stress response of Listeria monocytogenes varies between a strain of serotype 1/2a and a strain of serotype 4c. AB - The ability of Listeria monocytogenes to resist many adverse environmental conditions has been attributed in part to activation of the alternative sigma factor sigma(B), encoded by the sigB gene. The ability of this pathogen to survive and grow under stress conditions varies between strains within the species. The current study was undertaken to determine whether the role played by the sigB gene in the stress response varies among strains of different serotypes. Null mutations were generated in the sigB genes of L. monocytogenes L61 (serotype 1/2a) and L99 (serotype 4c), and the survival of the resulting mutants was compared with that of the wild-type strains under osmotic, oxidative, and carbon starvation stress conditions and on exposure to bacteriocins, ethanol, acid, and heat. Except in a few cases, strain L61 displayed greater dependence on the sigB products for survival of adverse conditions than did strain L99. The results of this study indicated that the relative importance of the sigB gene in the stress response is not the same in all strains of L. monocytogenes, and this difference may be specific to serotype groupings within the species. PMID- 12732956 TI - Heat-resistance and heat-shock response in the nosocomial pathogen Enterococcus faecium. AB - We have characterized the heat-shock response of the nosocomial pathogen Enterococcus faecium. The growth of E. faecium cells was analyzed at different temperatures; little growth was observed at 50 degrees C, and no growth at 52 degrees C or 55 degrees C. In agreement, a marked decrease of general protein synthesis was observed at 52 degrees C, and very light synthesis was detected at 55 degrees C. The heat resistance of E. faecium cells was analyzed by measuring the survival at temperatures higher than 52 degrees C and, after 2 h of incubation, viable cells were still observed at 70 degrees C. By Western blot analysis, two heat-induced proteins were identified as GroEL (65 kDa) and DnaK (75 kDa). Only one isoform for either GroEL or DnaK was found. The gene expression of these heat-shock proteins was also analyzed by pulsed-labeled experiments. The heat-induced proteins showed an increased rate of synthesis during the first 5 min, reaching the highest level of induction after 10 min and returning to the steady-state level after 20 min of heat treatment. PMID- 12732957 TI - Generation of novel plasmids in Escherichia coli S17-1(pSUP106). AB - When the highly metal-resistant acidophilic heterotrophic strain, Acidiphilium symbioticum KM2, was incubated with two Escherichia coli strains, viz. S17-1 (pSUP106) and K12, on a medium that supported growth of these two divergent species of different habitats, E. coli transconjugants were isolated that contained novel plasmids and were resistant to Zn(2+) (48 m M), Cu(2+) (12 m M), Ni(2+) (12 m M), chloramphenicol (50 microg/ml), and tetracycline (25 microg/ml). The transconjugant plasmids did not hybridize with any of the A. symbioticum KM2 plasmids. After curing of the plasmids, the transconjugants became sensitive to 12 m M Zn(2+), 12 m M Cu(2+), and 12 m M Ni(2+), but remained chloramphenicol and tetracycline resistant-the phenotypic markers that were originally present in pSUP106. That a part of pSUP106 was integrated into the chromosome of the transconjugants was evident from the hybridization of pSUP106 with chromosomal DNA of the cured derivatives of the transconjugants. Further, the transconjugant plasmids hybridized only with the chromosomal DNA of E. coli S17-1 and not with the chromosomal DNA of A. symbioticum KM2 or E. coli K12, suggesting their host chromosomal origin. Thus, the present study describes a unique event of genetic rearrangements in the E. coli strain S17-1 (pSUP106), resulting in the formation of novel plasmids conferring metal-resistance phenotypes in the cell. PMID- 12732958 TI - Characterization of a new Pseudomonas aeruginosa strain NJ-15 as a potential biocontrol agent. AB - Phylogenetic characterization of soil isolate NJ-15, based on sequence homology of a partial 746-bp fragment of 16SrDNA amplicon, with the ribosomal database sequences (http://www.msu.edu/RDP/cgis/phylip.cgi), validated the strain as Pseudomonas aeruginosa. The strain NJ-15 produced a substantial amount of indole acetic acid (IAA) in tryptophan-supplemented medium. Besides, the strain also exhibited significant production of both the siderophore and hydrogen cyanide (HCN) on chrome azurol S and King's B media, respectively. The data revealed lower HCN production under iron-limiting conditions vis-a-vis higher HCN release with iron stimulation. Significant growth inhibition of phytopathogenic fungi occurred in the order as Fusarium oxysporum > Trichoderma herizum > Alternaria alternata > Macrophomina phasiolina upon incubation with strain NJ-15 cells. Thus, the secondary metabolites producing new Pseudomonas aeruginosa strain NJ-15 exhibited innate potential of plant growth promotion and biocontrol activities in vitro. PMID- 12732959 TI - Nitrite as an energy-conserving electron sink for the acetogenic bacterium Moorella thermoacetica. AB - Nitrite served as an energy-conserving electron acceptor for the acetogenic bacterium Moorella thermoacetica. Growth occurred in an undefined (0.1% yeast extract) medium containing 20 m M glyoxylate and 5 m M nitrite and was essentially equivalent to that observed in the absence of nitrite. In the presence of nitrite, acetate (the normal product of glyoxylate-derived acetogenesis) was not detected during growth. Instead, growth was coupled to nitrite dissimilation to ammonium, and acetogenesis was limited to the stationary phase. Furthermore, membranes from glyoxylate-grown cells under nitrite dissimilating conditions were deficient in the b-type cytochrome that is typically found in the membranes of acetogenic cells. Unlike glyoxylate, other acetogenic substrates (fructose, oxalate, glycolate, vanillin, and hydrogen) were not growth supportive in the undefined medium containing nitrite, and glyoxylate dependent growth did not occur in a nitrite-supplemented, basal (without yeast extract) medium. Glyoxylate-dependent growth by Moorella thermoautotrophica was not observed in the undefined medium containing nitrite. PMID- 12732960 TI - Detection of an intracellular protease inhibitor in archaea. AB - Proteolytic activity and a subtilisin inhibitor (NSI) were detected in Natrialba magadii cells. The proteolytic activity was due to two different proteases: a approximately 90-kDa metallo protease (NMP) produced during exponential growth and a 246-kDa serine protease (NSP) detected in the stationary phase. Both proteases were detected in the cytosolic fraction. NSI activity was maximal during early stages of growth and decreased in the stationary phase. NSI is a 35 kDa thermosensitive protein; it inhibits NSP activity but has no effect on NMP, and it was detected as a soluble or membrane-bound protein depending on the growth phase. Our results suggest that NSI may regulate NSP activity in vivo and that this protease may have a role in stationary phase cells. To our knowledge, this is the first report on the occurrence of protease inhibitors in Archaea. PMID- 12732961 TI - Cloning of the Pichia anomala SEC61 gene and its expression in a Saccharomyces cerevisiae sec61 mutant. AB - In several organisms, including Saccharomyces cerevisiae and other yeast species, the product encoded by the SEC61 gene is considered to be the core element of the translocation apparatus within the endoplasmic reticulum membrane through which translocation of secretory and membrane proteins occurs. In this study, we have cloned and characterized the homolog of the SEC61 gene from the yeast Pichia anomala. The cloned gene includes an ORF, interrupted after the first ten nucleotides by an intron of 131 bp, encoding a 479-amino acid putative polypeptide exhibiting homology to the products encoded by different eukaryotic SEC61 genes, particularly to those from other yeast species. We show that the P. anomala SEC61 gene is correctly processed (intron splicing) when expressed in S. cerevisiae and that it is able to complement the thermosensitive phenotype associated with a mutation in the S. cerevisiae SEC61 gene. PMID- 12732962 TI - Neurotoxin gene clusters in Clostridium botulinum type A strains: sequence comparison and evolutionary implications. AB - The nucleotide sequence of the hemagglutinin ( ha) genes and the transcriptional regulator botR gene were determined in type A Clostridium botulinum strain 62A, and the complete nucleotide sequence of the botulinum neurotoxin (BoNT) gene cluster was determined in strain Hall A- hyper. Comparison of the BoNT/A gene clusters revealed only two nucleotide differences between the two strains. The nucleotide sequences of the regions flanking the BoNT clusters were also determined in strains 62A, Hall A- hyper, and type A(B) strain NCTC 2916. The regions upstream of the BoNT/A clusters in the type A strains shared marked homology with the region upstream of the silent BoNT/B cluster in the A(B) strain, indicating a similar evolutionary origin. The region downstream of the BoNT/A cluster in type A strains encodes putative insertion sequence (IS) elements with multiple internal mutations. These IS elements may have played a role in neurotoxin gene transfer within the host genome and to other Clostridium species. PMID- 12732963 TI - A mammalian cell regulatory agent, CeReS-18, inhibits yeast cell proliferation but not bacterial replication. AB - A cell regulatory sialoglycopeptide, CeReS-18, purified from intact bovine cerebral cortex cells, has exhibited the capability of reversibly inhibiting cellular DNA synthesis and the proliferation of a wide array of mammalian cells. In the present study, the effect of CeReS-18 on the proliferation of bacterial ( Bacillus cereus and Escherichia coli) and yeast ( Saccharomyces cerevisiae and Schizosaccharomyces pombe) cells was investigated. The results showed that replication and viability of the bacterial cells were not affected by CeReS-18 at any concentration tested, including 15-fold higher than that used for inhibiting mouse 3T3 cell proliferation. In contrast to bacterial cells, CeReS-18 was able to inhibit the replication of yeast cells, in a concentration-dependent, reversible manner, and the addition of calcium to the culture medium could abrogate the inhibitory effect of CeReS-18. A cytotoxic effect of CeReS-18 on both yeast cell species was observed when it was applied at higher concentrations. PMID- 12732964 TI - Genotypic diversity of oscillatoriacean strains belonging to the genera Geitlerinema and Spirulina determined by 16S rDNA restriction analysis. AB - Genotypic diversity of several cyanobacterial strains mostly isolated from marine or brackish waters, belonging to the genera Geitlerinema and Spirulina, was investigated by amplified 16S ribosomal DNA restriction analysis and compared with morphological features and response to salinity. Cluster analysis was performed on amplified 16S rDNA restriction profiles of these strains along with profiles obtained from sequence data of five Spirulina-like strains, including three representatives of the new genus Halospirulina. Our strains with tightly coiled trichomes from hypersaline waters could be assigned to the Halospirulina genus. Among the uncoiled strains, the two strains of hypersaline origin clustered together and were found to be distant from their counterparts of marine and freshwater habitat. Moreover, another cluster, formed by alkali-tolerant strains with tightly coiled trichomes, was well delineated. PMID- 12732965 TI - Polymorphism in the yclC-rpoS region of enterobacteria. AB - The nucleotide sequence downstream from the rpoS gene in Enterobacter cloacae and Kluyvera cryocrescens contains the slyA-pad1-yclC genes. The DNA sequence of Enterobacter cloacae CETC960 shows a 2.6-kb insertion of unknown origin between rpoS and slyA. This 2.6-kb sequence has also been detected in species of Salmonella and in Pseudomonas aeruginosa, but not in the same location. This insertion has been detected in all the Enterobacter cloacae clinical strains studied although the size of the rpoS-yclC region was highly variable, possibly owing to the presence of insertions and/or deletions. The study of the rpoS-mutS region in other enterobacteria also showed variability in size. Our results support the idea of a variational hot spot in the rpoS-mutS region that could be related to pathogenesis and horizontal transfer. PMID- 12732966 TI - Function of a low molecular weight peptide from Trichoderma pseudokoningii S38 during cellulose biodegradation. AB - The biochemical mechanism for cellulose decomposition by a low molecular weight peptide, named short fiber generating factor (SFGF), derived from the culture supernatant of a cellulolytic fungus Trichoderma pseudokoningii S-38, was determined. Sufficient information obtained by biochemical and biophysical studies and combined with observation with a scanning electron microscope provided further evidence for the earlier studies that the SFGF had a high capacity for chelating and reducing ferric ions, and could produce free radical by reduction of Fe(3+) to Fe(2+) in the presence of oxygen molecule. These studies suggested that the effect of SFGF on cellulose is directly related to an oxidative reaction and is different from the hydrolysis of cellulose by cellulases. The alcoholic hydroxyl groups in cellulose can be oxidized by SFGF, which leads to destruction of the hydrogen bond network in cellulose and cleavage of glycosidic linkages. Both effects led to the de-polymerization of cellulose and the formation of short fibers, and increase of reducing groups in residual cellulose, then the cellulose substrates became more susceptible for hydrolysis by cellulases. PMID- 12732967 TI - Inactivation of cyanobacterial nitrogenase after exposure to ultraviolet-B radiation. AB - Exposure of the N(2)-fixing cyanobacterium Anabaena BT2 to ultraviolet-B radiation (2.5 W m(-2)) for 30 min resulted in complete loss of nitrogenase activity but 100% cell killing occurred only after a 90-min exposure. Inactivation of nitrogenase activity was not specific to Anabaena BT2; other species also showed a similar effect. The time required for 100% killing and inactivation of nitrogenase activity differed in various species, and this difference may be ascribed to the presence of different levels of UV-B protection mechanisms in individual species. Inhibition of nitrogenase activity was immediate, since exposure of cultures to UV-B for as little as 5 min elicited some inhibition of activity. The activity of UV-B-inhibited nitrogenase did not recover upon transfer of exposed cells to fluorescent light, suggesting that the inhibition may be due to specific inactivation of the enzyme. By employment of inhibitors of protein synthesis and PS-II activity, it was demonstrated that restoration of nitrogenase activity in a UV-B-treated culture occurred by fresh synthesis of nitrogenase polypeptide. Our findings suggest that estimation of nitrogenase activity in diazotrophic species may be used as a marker enzyme for assessing the impact of UV-B radiation. PMID- 12732968 TI - Identification and characteristics of nisin Z-producing Lactococcus lactis subsp. lactis isolated from Kimchi. AB - We isolated bacteriocin-producing Lactococcus lactis subsp. lactis from Kimchi. The bacteriocin inhibited strains of Clostridium perfringens, C. difficile, Listeria monocytogenes, vancomycin-resistant Enterococcus, and one out of four methicillin-resistant Staphylococcus aureus strains, as well as some closely related lactic acid bacteria. In tricine-SDS-PAGE, the bacteriocin migrated with an apparent molecular weight of about 4 kDa to the same location as nisin A and crude nisin Z. The gene encoding this bacteriocin was found to be identical to that of nisin Z with direct PCR sequence methods. The inhibitory activity was stable against heat and pH, but it was lost at 100 degrees C for 1 h and at 121 degrees C for 15 min. The bacteriocin was inactivated by proteolytic enzymes, but was not affected by lysozyme, lipase, catalase, or beta-glucosidase. There were some differences in characteristics from those of nisins described previously. PMID- 12732969 TI - Genetic organization of genes encoding phenol hydroxylase, benzoate 1,2 dioxygenase alpha subunit and its regulatory proteins in Acinetobacter calcoaceticus PHEA-2. AB - Acinetobacter calcoaceticus PHEA-2 is a phenol-degrading bacterium isolated from the wastewater from an oil refinery. A 10-kb XhoI fragment consisting of nine complete Open Reading Frames (ORFs) and one partial ORF was screened from a lambda library of PHEA-2 by Southern hybridization. The sequence analyses revealed that ORF2-ORF7, designated mphKLMNOP, are homologous to dmpKLMNOP of Pseudomonas sp. CF600 and mopKLMNOP of Acinetobacter calcoaceticus NCIB8250, sharing 38%-72% and 58.5%-93.5% respectively. The products encoded by dmp and mop genes convert phenol to catechol. The mph-operon and downstream ORFs, ORF9 and ORF10, sharing high identities to benM and benA, which encode ben-operon regulatory protein and benzoate 1,2-dioxygenase alpha subunit respectively, are separated by ORF8, whose function is unknown. The organization of the mph and ben operons is different from that described previously. PMID- 12732970 TI - Type III secretion homologs are present in Brucella melitensis, B. ovis, and B. suis biovars 1, 2, and 3. AB - Protein sequences from characterized type III secretion (TTS) systems were used as probes in silico to identify several TTS gene homologs in the genome sequence of Brucella suis biovar 1 strain 1330. Four of the genes, named flhB, fliP, fliR, and fliF on the basis of greatest homologies to known flagellar apparatus proteins, were targeted in PCR and hybridization assays to determine their distribution among other Brucella nomen species and biovars. The results indicated that flhB, fliP, fliR and fliF are present in Brucella melitensis, Brucella ovis, and Brucella suis biovars 1, 2 and 3. Similar homologos have been reported previously in Brucella abortus. Using RT-PCR assays, we were unable to detect any expression of these genes. It is not yet known whether the genes are the cryptic remnants of a flagellar system or are actively involved in a process contributing to pathogenicity or previously undetected motility, but they are distributed widely in Brucella and merit further study to determine their role. PMID- 12732972 TI - Compositional difference of the exopolysaccharides produced by the virulent and virulence-deficient strains of Xanthomonas oryzae pv. oryzae. AB - Xanthomonas oryzae pv. oryzae, the bacterial blight pathogen of rice, is known to produce phytotoxic polysaccharides. The extracellular polysaccharide (EPS) was isolated from virulent (BXO1) and virulence-deficient gum G mutant (BXO1002) strains of X. oryzae pv. oryzae and characterized using fourier transform infrared (FT-IR) and nuclear magnetic resonance (NMR). Data from the FT-IR suggested that the aldehyde (R-CHO) group and C=O of acid anhydride are present in BXO1 but absent in BXO1002. The (1)H-NMR spectra showed the presence of an acetyl amine of hexose or pentose, free amines of glucose, an beta-anomeric carbon of hexose and pentose, hydrogen next to hydroxyl group, an acetyl amine of hexose and pentose in the polysaccharides of both BXO1 and BXO1002, and the absence of alpha-anomeric carbon of hexose or pentose and the glucuronic acid in the polysaccharides produced by BXO1002. The test for glucuronic acid also confirmed the absence of glucuronic acid in the polysaccharides of BXO1002 and the presence glucuronic acid (32 microg/mg) in the polysaccharides produced by BXO1. PMID- 12732971 TI - Studies on the expression of regulatory locus sae in Staphylococcus aureus. AB - Global regulatory locus sae consists of a two-component signal transduction system coded by saeR and saeS genes that upregulates the transcription of several exoproteins. Northern analysis carried out in this study reveals the synthesis at late and post-exponential phases of a cotranscript of saeR and saeS structural genes of about 2.4 kb. This transcript is diminished in the isogenic agr:: tetM mutant. Likewise, transcriptional fusion experiments show that sae expression is downregulated in the agr null mutant. Complementation analyses with plasmids carrying fragments of about 1.2 or 0.2 kbp upstream of saeR-saeS genes, which restore fully or only partially, respectively, the wild-type phenotype to the sae mutant, are in agreement with two initiation start points of transcription revealed by primer extension experiments. This work, as well as previous studies, reveals a complex hierarchical regulatory network involving several loci that control the expression of virulence determinants in S. aureus. PMID- 12732973 TI - Molecular typing by pulsed-field gel electrophoresis of Bacillus thuringiensis from root voles. AB - The root voles intestinal strains of Bacillus thuringiensis, were characterised by pulsed-field gel electrophoresis (PFGE). For 14 isolates, three pulsotypes were found, with the use of SmaI or NotI as restriction enzymes. Strains in each pulsotypes presented identical DNA patterns, indicating that the population structure of B. thuringiensis from root voles is clonal. The similarities in banding patterns were estimated at 56% and 33% for SmaI and NotI digests, respectively. The strains under study differed significantly in the size of their entire genome, which varied between 2.4 and 4.2 Mb. No significant differences were detected among the isolates subjected to biochemical properties determined by API tests. Present study showed that genomic diversity is a common feature of B. thuringiensis originating from one ecological niche. PFGE appears to be a useful technique for use in studies on the spread of B. thuringiensis in the environment. PMID- 12732974 TI - Chemotactic behavior of Chlamydomonas reinhardtii is altered during gametogenesis. AB - Chemotactic behavior of Chlamydomonas reinhardtii is altered during the sexual life cycle. Unlike vegetative cells and noncompetent pregametes, mature gametes did not show chemotaxis to ammonium. Loss of chemotaxis to ammonium in mating competent cells is controlled by gamete-specific genes that are common for both mating-type gametes. Change of chemotaxis mode requires the sequential action of the two environmental signals: removal of ammonium from the medium and light. The mutants lrg1, lrg3, and lrg4 affected in the light-dependent step of sexual differentiation exhibited the loss of chemotaxis to ammonium in the absence of light. These data indicate that there are common components in the signaling pathways that control change of chemotactic behavior and forming of mating competence in gametes. PMID- 12732975 TI - Ca2+ -dependence and inhibition of transformation by trifluoperazine and chlorpromazine in Thermoactinomyces vulgaris. AB - Ca(2+) enhanced the transformation frequency of Thermoactinomyces vulgaris (stock no. 1278) of an auxotrophic strain by the chromosomal DNA isolated from a prototrophic strain (stock no. 1227). The number of transformants showed a marked increase with increasing concentration of CaCl(2) upto 0.05 mM; and above this concentration, the transformation frequency decreased significantly. Antipsychotic drugs that are potent calmodulin inhibitors, like trifluoperazine and chlorpromazine, when applied in the concentration range of 0.01-0.04 mM along with optimal CaCl(2) concentration to the cultures of the recipient cells, resulted in a significant inhibition in the frequency of Ca(2+)-stimulated transformation. The results of present investigation suggest the involvement of a Ca(2+)-dependent protein activator in the development of Ca(2+)-mediated competence, which could have played an important role in the enhancement of genetic transformation in this aerobic spore forming thermophilic actinomycete. PMID- 12732976 TI - The role of cell hydrophobicity in the formation of aerobic granules. AB - Cell hydrophobicity is an important affinity force in cell self-immobilization and attachment processes. The role of cell hydrophobicity in the formation of aerobic granules has not been clear. Therefore, two series of experiments were conducted to investigate the role of cell hydrophobicity in the formation of aerobic heterotrophic and nitrifying granules in sequencing batch reactors, while the effects of shear strength, hydraulic selection pressure, and organic loading rate on the cell hydrophobicity were also studied. Results showed that the formations of heterotrophic and nitrifying granules were associated very closely with the cell hydrophobicity. The hydrophobicity of granular sludge was nearly twofold higher than that of conventional bioflocs. A high shear force or hydraulic selection pressure imposed on microorganisms resulted in a significant increase in the cell hydrophobicity, while the cell hydrophobicity seemed not to be sensitive to the changes in the organic loading rates in the range studied. In conclusion, the cell hydrophobicity could induce and further strengthen cell-cell interaction, and might be a main triggering force to initiate the granulation of heterotrophic and nitrifying bacteria. PMID- 12732977 TI - Carbon and nitrogen source nutrition of fumagillin biosynthesis by Aspergillus fumigatus. AB - The carbon and nitrogen source requirements of Aspergillus fumigatus NRRL 2436 for growth and production of the angiogenesis inhibitor fumagillin were studied in chemically defined media. Both carbon and nitrogen sources strongly influenced fumagillin formation. Two out of 29 carbon sources tested interfered with fumagillin biosynthesis. The best combination of two carbon sources was 30 g L( 1) xylan and 50 g L(-1) mannose. Of fifteen nitrogen sources tested, three ammonium salts (chloride, sulfate, and dibasic phosphate) failed to support fumagillin formation, presumably due to the low pH which developed. The dosage response study of the best nitrogen source, L-glutamic acid, revealed that 9 g L( 1) was optimal. Volumetric production of fumagillin was increased by 15-fold over that in the starting (Peterson-Goldstein) medium as a result of these findings. PMID- 12732978 TI - Cell surface hydrophobicity and slime production of Staphylococcus epidermidis Brazilian isolates. AB - The cell surface hydrophobicity of 60 isolates and three reference strains of Staphylococcus epidermidis was assayed by means of bacterial aggregation in liquid broth, phosphate-buffered saline, and in ammonium sulfate, as well as by affinity of the bacteria to n-hexadecane and polystyrene surfaces. In order to better characterize the isolates, the influence of bacterial growth time and enzyme treatment on cell hydrophobicity and the analysis of the slime production were also investigated. The strains presented the following profiles when assayed by the ammonium sulfate aggregation test (SAT): SAT < 1M, SAT 1M - <2M, SAT 2M - <4M, and SAT >or=4M. When SAT < 1M, the strains showed positive results for most of the cell surface hydrophobicity tests. None of the strains belonging to the groups with SAT >or= 1M showed spontaneous aggregation (SA), auto-aggregation (AA), or glass adherence, albeit 32 (62.7%) strains were polystyrene adherent and 42 (82.3%) presented weak adherence to n-hexadecane (>20%). The best correlation of the results was found among the AA and glass adherence tests (100%), followed by SA/ glass adherence (98%) and SA/ AA test (98%). The polystyrene adherence test and microbial adherence to n-hexadecane test (MATH) showed 78% correlation. Proteinase K treatment reduced bacterial adherence to polystyrene, but did not influence the SAT values. Three distinct groups of strains were distinguished by the polystyrene micromethod and glass tube adherence assay: 0.0-0.4 O.D. group, including non-glass adherent isolates; 0.5-0.7 O.D. group, including strains with variable profiles (adherent or non-adherent); and 0.8-1.3 O.D. group, composed of glass-adherent strains. Evaluation by a single method seemed not to reliably determine the surface hydrophobicity characteristics of S. epidermidis clinical isolates. Auto-aggregation properties of the strains that adhered to glass seemed related to slime expression, rather than cell surface hydrophobicity. Data also suggested involvement of protein components in adherence to polystyrene, but not in auto-aggregation properties assayed by SAT. PMID- 12732979 TI - Characterization of flagellar antigens and insecticidal activities of Bacillus thuringiensis populations in animal feces. AB - In total, 287 Bacillus thuringiensis isolates, recovered from feces of 28 zoo maintained animal species, were examined for flagellar (H) antigenicity and insecticidal activity. Serologically, 209 isolates (72.8%) were allocated to the 8 H serogroups, 4 were untypable, and 74 were untestable. Among the 8 H serotypes detected, H3abc (serovar kurstaki) predominated at a high frequency of 88.0%, followed by H6 (serovar entomocidus) with a frequency of 7.7%. Insecticidal activity was associated with 67.2% of the fecal populations: 188 isolates were toxic to both Bombyx mori (Lepidoptera: Bombycidae) and Aedes aegypti (Diptera: Culicidae), 2 isolates were specific for B. mori, and 3 isolates were toxic to A. aegypti only. Of the isolates with dual toxicity, 97.9% belonged to the serovar kurstaki, producing bipyramidal parasporal inclusions. All of the H7 (serovar aizawai) isolates were toxic to both insects. PMID- 12732980 TI - Gene cloning, sequencing, and expression of an esterase from Acinetobacter lwoffii I6C-1. AB - The esterase-encoding gene, estA, was cloned from Acinetobacter lwoffii I6C-1 genomic DNA into Escherichia coli BL21(DE3) with plasmid vector pET-22b (pEM1). pEM1 has a 4.4-kb EcoRI insert that contained the complete estA gene. A 2.4-kb AvaI- SphI DNA fragment was subcloned (pEM3) and sequenced. estA gene encodes a protein of 366 amino acids (40,687 Da) with a pI of 9.17. The EstA signal peptide was 31 amino acids long, and the mature esterase sequence is 335 amino acids long (37.5 kDa). The conserved catalytic serine residue of EstA is in position 210. The EstA sequence was similar to that of the carboxylesterase from Acinetobacter calcoaceticus (75% identity, 85% similarity), Archaeoglobus fulgidus (37% identity, 59% similarity), and Mycobacterium tuberculosis (35% identity, 51% similarity). These enzymes contained the conserved motif G-X(1)-S-X(2)-G carrying the active-site serine of hydrolytic enzyme. The EstA activity in A. lwoffii I6C 1 remains constant throughout the stationary phase, and the activity in E. coil BL21 (DE3) with pEM1 was similar to A. lwoffii I6C-1. PMID- 12732981 TI - Biochemical and functional characterization of a eukaryotic-type protein kinase, SpkB, in the cyanobacterium, Synechocystis sp. PCC 6803. AB - On the basis of the genome sequence, the unicellular motile cyanobacterium Synechocystis sp. PCC 6803 harbors seven putative genes for eukaryotic-type protein kinase belonging to Pkn2 subfamily ( spkA approximately spkG). Previously, SpkA was shown to have protein kinase activity and to be required for cell motility. Here, the role of the spkB was examined. The spkB gene was expressed in Escherichia coli as a fusion protein with His-tag, and the protein was purified by Ni(2+) affinity chromatography. The eukaryotic-type protein kinase activity of the expressed SpkB was demonstrated as autophosphorylation to itself and phosphorylation of the general substrate proteins. SpkB showed autophosphorylation activity in the presence of both Mg(2+) and Mn(2+), but not in Ca(2+). Phenotype analysis of spkB disruptant of Synechocystis revealed that spkB is required for cell motility, but not for phototaxis. These results suggest that SpkB is the eukaryotic-type protein kinase, which regulates cellular motility via protein phosphorylation like SpkA. PMID- 12732982 TI - Characterization of an IS element in Mycoplasma orale that is highly homologous to M. fermentans ISLE. AB - In a previous study, using a primer set designed from Mycoplasma fermentans, we amplified a PCR fragment from Mycoplasma orale similar to the 206-bp DNA fragment amplified from M. fermentans insertion-sequence-like element (ISLE). The presence of this similar ISLE fragment has the potential to cause confusion in the PCR diagnosis of M. fermentans and M. orale, which have significantly different clinical scenarios. An ISLE from three different M. orale strains was amplified by using a primer set designed from sequence within the left and right terminal stem and loop (S&L) structures flanking the ISLE of M. fermentans. Sequence analysis showed that the M. orale ISLE is 93% identical to that of M. fermentans at the nucleotide level and codes for two open reading frames also found in the M. fermentans ISLE. This is the first finding that two different mycoplasma species harbor highly homologous IS elements. This finding has great significance in clinical diagnosis and suggests a possibility of horizontal transfer of an IS element between two different mycoplasma species. PMID- 12732983 TI - The non-catalytic amino acid Asp446 is essential for enzyme activity of the modular endocellulase Cel9 from Myxobacter sp. AL-1. AB - The modular endocellulase Cel9 of the bicistronic operon cel9-cel48 of Myxobacter sp. AL-1 shares not only amino acid sequence similarity but also biochemical properties similar to those of Thermobifida fusca endo/exocellulase E4. Amino acid alignments of a T. fusca E4 cellulase subfamily of family 9 cellulases revealed that Asp(446) of Myxobacter sp. AL-1 Cel9, a putatively noncatalytic residue, is highly conserved in one of the catalytic domains of this subfamily. Directed mutagenesis of residue aspartate (Asp(446)) to alanine generated a Cel9 mutant that lost more than 99% of its activity, suggesting that Asp(446) plays an essential structural role in Cel9 during cellulose degradation. Owing to its high degree of conservation and essential role, we propose that Asp(446) of Myxobacter sp. AL-1 Cel9 is a good landmark that distinguishes members of the E4 subfamily of family 9 cellulases. PMID- 12732984 TI - Diagnosis and management of primary aldosteronism. AB - Identifying primary aldosteronism within the hypertensive population is an important clinical challenge, as most patients with a unilateral source of excess aldosterone secretion are amenable to surgical cure. At least 20% of patients with primary aldosteronism have normal serum potassium levels. Therefore, screening tests should not be based on recognition of hypokalemia alone. Rather, the diagnosis should depend on identifying renin suppression and measuring the ratio of plasma aldosterone concentration to plasma renin activity. The diagnosis may be confirmed by performing an aldosterone suppression test after oral salt loading. Once primary aldosteronism has been established, it is necessary to exclude glucocorticoid-remediable aldosteronism and then proceed to localization studies. Detecting a unilateral source of aldosterone, usually due to an adenoma (Conn syndrome), is achieved by postural hormonal testing and confirmed by selective venous sampling (SVS) with measurement of aldosterone concentrations (expressed as the aldosterone/cortisol ratio) in each adrenal vein. SVS is enjoying a revival in many institutions as it is more sensitive and specific than either cross-sectional imaging or scintigraphy and has the potential to influence significantly both the diagnosis and clinical decision-making. Patients with unilateral disease are ideally treated by laparoscopic adrenalectomy. Patients in whom localization is not achieved usually have bilateral adrenal hyperplasia and are treated medically. PMID- 12732985 TI - A decreasing trend in biliary output in fistulas is more important than cutoff value or duration. PMID- 12732986 TI - Plastic iodophor drape during liver surgery operative use of the iodophor impregnated adhesive drape to prevent wound infection during high risk surgery. AB - We retrospectively investigated factors associated with wound infection after liver resection for hepatocellular carcinoma (HCC), with special reference to use of a plastic adhesive drape impregnated with iodophor. The subjects were 296 patients undergoing liver resection for HCC. Wound infection was defined as purulent drainage from the superficial incision with or without laboratory confirmation. One or more of the following signs was required: pain or tenderness, localized swelling, or redness or heat. Wound infection developed in 25 patients. Regression analysis indicated that low body mass index (BMI), smoking, long preoperative hospital stay, and nonuse of iodophor drapes were risk factors for wound infection. Wound infection was significantly less likely with the use of iodophor drapes (3.1%) than for surgery without iodophor drapes (12.1%). By multivariate regression analysis, BMI, smoking, and lack of drape use were independent risk factors. Most of the bacteria isolated were skin bacteria, including Staphylococcus aureus and Staphylococcus epidermidis. In conclusion, low BMI, smoking, a long preoperative hospital stay, and the lack of iodophor drape use were risk factors for wound infection after liver resection for HCC. The drapes presumably prevented contamination from the skin during the operation. PMID- 12732988 TI - Benefit of rFVIIa administration for cirrhotic patients undergoing surgery. PMID- 12732989 TI - Carl Gussenbauer: pioneer in pancreatic surgery. AB - Carl Gussenbauer (1842-1903), known for many contributions to surgery, should also be noted prominently in the list of pioneers of pancreatic surgery. His successful performance of external drainage for a pancreatic cyst in 1882 was the first planned pancreatic operation. Although pancreatic surgery had been developed in animal models during the seventeenth century, surgery of the human pancreas was practically nonexistent owing to the dismal results seen with incidental pancreatic operations done early in the nineteenth century. Gussenbauer's procedure was a significant advance, representing a safe and effective solution to the pancreatic cyst problem. Gussenbauer's external drainage procedure remained the gold standard for treating pancreatic cysts until the 1920s and was still a widely practiced therapeutic option until the 1950s. With his discipline, insight, and innovative thought, Gussenbauer initiated the modern era of pancreatic surgery. PMID- 12732987 TI - Change in mitochondrial membrane potential in peripheral blood lymphocytes, especially in natural killer cells, is a possible marker for surgical stress on the immune system. AB - There is accumulating evidence that surgical stresses cause impairment of systemic immune responses, which may promote susceptibility to infection as well as growth of remnant cancer cells in cancer patients. Although alterations in numbers, populations, and functions of lymphocytes have been extensively studied to assess modulation of the immune system, the precise mechanisms of immunosuppression caused by surgical stresses have not been identified, nor have methods been developed to estimate the magnitude of surgical stresses on the immune system. In the present study, to evaluate the effects of surgical procedures on the immune system, the mitochondrial membrane potential (Delta Psi(m)) of peripheral blood lymphocytes (PBL) from 25 patients who underwent various types of operation was measured by flow cytometry using 3,3' dihexiloxacarbocyanine iodide (DiOC(6)(3)) on the day before operation and on postoperative day (POD) 1, POD 3, and POD 7. The Delta Psi(m) in PBL, especially in natural killer (NK) cell population, was reduced after major surgery. In particular, the reduction of Psi Delta(m) in NK cells appeared to be proportional to the severity of the surgical procedures and reflected the impairment of cellular function. Interestingly, the Delta Psi(m) in NK cells was also negatively correlated with the level of plasma noradrenaline after major surgery, suggesting that the reduction of Delta Psi(m) in NK cells induced by surgical stresses may be mediated, at least in part, by the accompanying increase in plasma noradrenaline. Monitoring of Delta Psi(m) in PBL after operation may be one of the useful markers for estimating the magnitude of surgical stresses on the immune system. PMID- 12732990 TI - Diagnosis and treatment of inferior vena caval invasion by hepatic cancer. AB - Hepatectomy with concomitant resection of the inferior vena cava (IVC) has become common for hepatic malignancies involving the IVC. However, diagnosing IVC invasion and the procedure of choice have yet to be standardized. Medical records of nine patients with liver cancer (five metastatic tumors from colorectal cancer and four intrahepatic cholangiocarcinomas) believed to have directly invaded the IVC wall were retrospectively abstracted for data on preoperative radiologic studies, surgical procedures, histology of the resected specimen, and treatment outcome. All nine patients underwent hepatectomy: Five did not undergo IVC resection because the IVC could be isolated from the tumor; the remaining four underwent combined IVC resection (wedge and segmental resections in two each). The segmentally resected IVC was reconstructed using an external iliac vein graft. Total hepatic vascular exclusion, venovenous bypass, and the ex vivo technique were not used. Interestingly, the tumor was smaller and the percentage of the IVC circumference in contact with tumor as seen on computed tomography (CT) was less in patients with IVC invasion than in those without it (40 +/- 11 vs. 134 +/- 61 mm, p < 0.05; 30% +/- 8% vs. 60% +/- 20%, p < 0.05). The length of the IVC compressed by tumor on cavography was similar in the two patient groups (47 +/- 9 vs. 55 +/- 8 mm). All patients were discharged from the hospital in good condition: Seven died of cancer recurrence, and the remaining two are currently alive and disease-free 15 and 73 months after surgery, respectively. In conclusion, imaging modalities demonstrating caval deformation, such as CT and cavography, are unreliable for diagnosing direct invasion of the IVC wall. Even when IVC invasion is strongly suggested by conventional radiologic studies, the surgeon should endeavor to peel the tumor from the IVC. This strategy is important to avoid unnecessary resection of the IVC, use of a prosthetic graft, or ex vivo hepatectomy. PMID- 12732991 TI - Risk factors and management of bile leakage after hepatic resection. AB - The aim of this study was to identify the perioperative risk factors for postoperative bile leakage after hepatic resection and to propose a treatment strategy for such leakage when it does occur. Between 1992 and 2000 a total of 313 hepatic resections without choledocojejunal anastomosis were performed at our institute. Risk factors related to bile leakage were identified with univariate analysis, and strategies were evaluated in relation to the findings of postoperative fistulography. Postoperative bile leakage developed in 17 patients (5.4%). Univariate analysis identified high risk factors as advanced age, a wide surface area of the incision (bile leakage group versus no bile leakage group: 102.1 vs. 66.4 cm(2), p < 0.05), and exposure of Glisson's sheath at the cut surface (e.g., central bisegmentectomy, S4, S8 subsegmentectomy). Groupings of patients by their postoperative fistulography results showed that patients with involvement of the proximal bile duct were slower to heal than those with no demonstrable bile duct involvement. The one patient whose fistulogram demonstrated peripheral bile duct involvement had uncontrollable leakage and required reoperation. Hepatectomies with a wide surface area and those that expose the major Glisson's sheath present serious risk factors for bile leakage. When the fistulogram shows proximal bile duct involvement, endoscopic nasobiliary tube drainage is necessary; when the fistulogram shows peripheral bile duct involvement, reoperation is needed. PMID- 12732992 TI - Nihilism: a benign denial. AB - Nihilism is the belief that all possible knowledge on a given topic has been amassed and codified. Ranging from benign denial to deliberate attempts at excommunication, nihilism is often encountered in the history of medicine. Eustachius, Columbus, and Sylvius strongly criticized Vesalius and defended the authority of Galen. Riolan fervently rejected Harvey's monumental work on the circulation of blood. Gross stated that no honest and sensible surgeon would ever sanction thyroidectomy. Sandstrom's discovery of the parathyroids was met with silence. Transplantation of parathyroids by Mandl was not appreciated when announced. Aristotle's dictum that the heart cannot withstand serious injury led to Paget's statement that cardiac surgery had reached the limits set by nature, which no new techniques could overcome. The first Billroth I operation was welcomed as, "Hopefully, also the last." Pancreatic surgery was opposed because the organ was of no clinical interest and was impossible for surgeons to reach. Pancreatic transplantation was rejected for many years, despite good results. When Blundell used blood transfusion for postpartum hemorrhage, critics averred that his next exploit would be radical removal of the spleen. Bassini stated that it could be risky to publish more about radical treatment of inguinal hernias. Carcinomas of the lower sigmoid and upper rectum were deemed untreatable because of their inaccessibility. Colostomy during pediatric surgery was rejected many times. Although it is difficult for the human mind to move from a familiar point of view, this propensity should not infect science, thereby impeding advancement. PMID- 12732993 TI - Chromoendoscopic evaluation of gastric mucosa after partial gastrectomy by use of modified endoscopic Congo red test. AB - The need for partial gastrectomy has decreased as a result of reduced incidence and improved endoscopic and medical treatment of peptic ulcer disease. Nonetheless, several patients with resected stomach remain in the population, and it is well known that important pathological changes can occur in the gastric remnant. We evaluated the morphological and functional status of the gastric stump by use of modified endoscopic Congo red test (MCRT). For this purpose, 87 partially gastrectomized (Billroth I and II) patients referred for elective gastroscopy were consecutively enrolled. We found a high prevalence of severe chronic atrophic fundal gastritis (CAFG) (67%) in the gastric remnant. We also observed, however, that one-third of the patients had almost unaffected gastric acid production even as long as 26 years after partial gastrectomy. Moreover, the accuracy, sensitivity, and specificity of routine gastroscopy in diagnosing CAFG in the gastric stump were found to be 55%, 50%, and 84%, respectively. The presence of bile reflux correlated well with the degree of CAFG. Importantly, we observed that more than 71% of the patients receiving acid-suppressing therapy had no or very little capacity to produce gastric acid. Taken together, our study has demonstrated that MCRT is a simple and well-tolerated method providing important morphological and functional information about the mucosa of the resected stomach. Furthermore, MCRT was superior to routine gastroscopy in diagnosing CAFG in the gastric stump. Outcome studies should define the clinical benefit of MCRT in the management of patients with resected stomach. PMID- 12732994 TI - Transthoracic versus transabdominal surgical approach for echinococcal cysts located over the superoposterior aspect of the right lobe of the liver. AB - A retrospective study of 80 patients operated on for hydatid cysts located on the superoposterior aspect of the right lobe of the liver (segments VI, VII, VIII) is presented. Right thoracotomy was performed in 30 patients, and 50 patients were operated on through bilateral subcostal incisions. The two approaches were compared in terms of radicality and morbidity. The transabdominal approach produced superior results and fewer postoperative complications, resulting in a shorter hospital stay (11 +/- 5 vs. 18 +/- 8 days). Total cystopericystectomy was feasible in 30% of patients operated on transabdominally and in 6% of those approached transthoracically. The rest of the patients were offered partial pericystectomy, except three in the thoracotomy group who underwent simple drainage of the cavity. We recommend that the transabdominal approach be the first choice for treatment of liver hydatid cysts irrespective of their location and size. We abandoned the transthoracic approach for cysts located on the superoposterior aspect of the right liver lobe in 1996. The transabdominal approach enables the surgeon to treat liver hydatidosis in a more radical, safer manner than does the transthoracic approach. PMID- 12732995 TI - Risk factors for nonhepatic surgery in patients with cirrhosis. AB - Cirrhosis of the liver appears to have an unfavorable prognosis in the surgical patient. The aim of this study was to determine risk factors for morbidity and mortality in patients with cirrhosis undergoing nonhepatic surgery. We studied 135 patients with liver cirrhosis undergoing nonhepatic procedures and 86 controls matched by age, sex, and preoperative diagnosis. Preoperative, intraoperative, and postoperative variables associated with 30-day mortality and morbidity were assessed by univariate and multivariate analyses. Patients with cirrhosis showed higher blood transfusion requirements, longer length of hospital stay, and a higher number of complications than controls. The mortality rate was 16.3% in cirrhotics and 3.5% in controls. By univariate analysis, the need for transfusions, prothrombin time, and Child-Pugh score were significantly associated with postoperative liver decompensation, whereas duration of surgery, prothrombin time, Child-Pugh score, cirrhosis-related complications, and general complications were significantly associated with mortality. In the multivariate analysis, Child-Pugh score (odds ratio [OR] 24.4; 95% confidence interval [CI] 5.5 to 106); duration of surgery (OR 5; 95% CI 1.2 to 15.6), and postoperative general complications (OR 3.7; 95% CI 3.4 to 6.4) were independent predictors of mortality. Patients with cirrhosis undergoing nonhepatic operations are at significant risk of perioperative complications leading to death. Independent variables associated with perioperative mortality include preoperative Child-Pugh score, the duration of surgery, and the presence of postoperative general complications. PMID- 12732997 TI - Laparoscopy-assisted repair of diaphragm injuries. AB - In this study we review our experience with an alternative laparoscopy-assisted technique for repair of diaphragm injuries. All patients admitted with an isolated diaphragm injury who underwent laparoscopy-assisted repair between January 1996 and February 2000 were included in the study. The diagnosis of the diaphragm injury was either obvious, with omentum herniating through the chest wall, or occult with confirmation of the injury at laparoscopy. Repair of the diaphragm was performed using standard surgical instruments via a 4-cm subcostal incision with use of abdominal wall traction and the laparoscope for visualization of the defect. A total of 24 patients underwent successful laparoscopy-assisted repair; There were twenty men and four women with an average age of 28.8 years (range 23-49 years). The average Revised Trauma score was 12. There were 23 grade II and 2 grade III diaphragm injuries. The mean operative time was 61 minutes (range 25-120 minutes) and the average hospital stay from the time of surgery to the time of discharge was 2.29 days (range 1-4 days). The procedure failed in one patient as a result of gaseous distension of the bowel. One patient developed a tension pneumothorax, and two others developed atelectasis. Laparoscopic-assisted repair of isolated diaphragm injuries using abdominal wall traction and standard surgical instruments is a feasible option with minimal morbidity and no mortality. PMID- 12732996 TI - Immunohistochemical evaluation for intraoperative rapid pathological assessment of the gastric margin. AB - In patients undergoing surgery for gastric cancer, as the resection area is difficult to define preoperatively, we define the resection area by intraoperative rapid pathological assessment of the resected margin. In some patients, however, the result of postoperative assessment of a permanent section differs from the result obtained intraoperatively. In this study we explored methods of improving the accuracy of intraoperative pathological assessment. Of the patients who underwent surgery for gastric cancer at Takarazuka Municipal Hospital, between April 2000 and July 2001, intraoperative pathological assessment of the resection margin was performed in 34 because it could not be accurately defined preoperatively. The intraoperative assessment consisted of touch smear cytological examination and hematoxylin and eosin (HE) and cytokeratin (CK) histological examination. The time required to perform cytological examination and HE and CK immunohistochemistry by this modified method, which uses an ENVISION Kit/HRP, was approximately 20 minutes for each procedure. Touch smear cytological examination gave false positive results in 3 of 34 patients (8.8%). Of the 14 patients preoperatively diagnosed with signet ring cell carcinoma, intraoperative HE histological analysis yielded a false negative result in one (7.1%). The results of CK histological assessment of these patients were all consistent with those of postoperative examination of permanent sections. Of the 20 patients preoperatively diagnosed with non-signet-ring cell carcinoma, HE analysis yielded a false negative result in one (5%). The results of CK histological assessment of these patients were all consistent with those of postoperative examination of permanent sections. Our modified CK staining method with the ENVISION Kit/HRP allows intraoperative pathological assessment to be performed accurately and rapidly. This method is more useful than HE immunohistochemistry for the assessment of resected gastric margins. PMID- 12732998 TI - Role of somatostatin in the prevention of pancreatic stump-related morbidity following elective pancreaticoduodenectomy in high-risk patients and elimination of surgeon-related factors: prospective, randomized, controlled trial. AB - A prospective, randomized, controlled trial was performed to determine the efficacy of somatostatin in the prevention of pancreatic stump-related complications with elimination of surgeon-related factors in high-risk patients undergoing pancreaticoduodenectomy. From August 1997 to December 2000, 54 patients, 28 men and 26 women, with age ranged from 32 to 89 years, were randomly assigned to somatostatin group ( n = 27) or placebo group ( n = 27). Ninety-four percent of the patients had pancreatic and periampullary lesions; 6% had secondary lesion involving the duodenum such as local recurrent colon carcinoma and renal cell carcinoma. These patients received either standard pancreaticoduodenectomy or pylorus-preserving pancreaticoduodenectomy. An experienced surgeon performed all operations in same fashion to minimize the surgical factor. A transanastomotic tube was inserted into the pancreatic duct for diversion of pancreatic juice in the pancreaticojejunostomy for a 3-weeks period postoperatively. Intravenous infusion of somatostatin was given at a dose of 250 microg/hr in the somastotatin group and normal saline was given to the control group for 7 days postoperatively. There was one perioperative death in each group, resulting in a 3.7% mortality rate. In the somastotatin group, as compared to the placebo group, the incidence of overall morbidity and pancreatic stump related complications were significantly lower with a mean decrease of 50% pancreatic juice output and a slightly shorter duration of hospital stays. In conclusion, after excluding surgeon related factor, prophylactic use of somatostatin reduces the incidence and severity of pancreatic stump related complications in high-risk patients having pancreaticoduodenectomy via decreased secretion of pancreatic exocrine. PMID- 12732999 TI - Acute compartment syndrome of the lower leg: retrospective study on prevalence, technique, and outcome of fasciotomies. AB - The acute compartment syndrome is caused by bleeding or edema in a closed muscle compartment surrounded by fascia and bone. It is characterized by increased intracompartmental pressure and decreased tissue perfusion. Well-known causative incidents are acute trauma and reperfusion after treatment for acute arterial obstruction. Most commonly the lower leg is involved. Inadequate therapy of the syndrome usually leads to muscle ischemia, rhabdomyolysis, and renal insufficiency. Perioperative morbidity and mortality are high. Although compartment syndromes can be caused by various factors, up until now no comparative studies have been published on clinical outcome of compartment syndromes of different origin. In this retrospective study we analyzed 40 successive cases of fasciotomy for acute lower leg compartment syndrome to study whether different causes of the syndrome lead to different clinical outcomes. We also studied other predictive factors for clinical outcome. The causes for the compartment syndromes were trauma, vascular deobstruction, cardiac surgery, and gastrointestinal surgery in lithotomy position. Clinical outcome showed a mortality of 15% and serious overall morbidity. Multivariate analysis showed the only significant predictive determinant of outcome to be the age of the patient. Fasciotomy for acute compartment syndrome is associated with serious morbidity and mortality. No correlation between causative factors and clinical outcome could be found. PMID- 12733000 TI - Resection of hilar cholangiocarcinomas: pivotal prognostic factors and impact of tumor sclerosis. AB - The well-known poor prognosis of proximal bile duct cancer is due to its unfortunate anatomical location and its late diagnosis. Successful tumor resection, which is considered to be optimal treatment, depends on many factors. Eighty-eight patients suffering from proximal bile duct cancer underwent surgical exploration at our institution between 1977 and 1998. In 37 patients the tumor was resectable; in the remaining 51 patients exploratory laparotomy or a palliative operation was performed. The median survival after tumor resection was 18.6 months, but median survival after a palliative procedure or an exploratory laparotomy was only 3.4 months (p < 0.001). A curative R0 resection was possible in 11 patients, an R1 resection was performed in 22 patients, and 4 patients had an R2 resection. The median survival rate after R0 resection was 83.6 months, 12.3 months after R1 resection, and 2.7 months after R2 resection (p < 0.001). Survival after resection in patients with negative lymph nodes (n = 30) was significantly longer than in those with positive lymph nodes (n = 7) (p = 0.022). Grade of tumor sclerosis tended to have an influence on resectability rate (p = 0.076). The pattern of tumor growth was without statistical influence. Multivariate analysis revealed resection (p < 0.001) as the only significant prognostic marker for patient survival. Radical resection is the only therapy that provides a chance for long-term survival, with sclerosis of the cancer tending to have an influence on univariate analysis. PMID- 12733001 TI - Liver trauma: experience in 348 cases. AB - Liver trauma, the main cause of death in patients suffering abdominal injury, remains an unresolved problem, especially in its most severe forms. The objective of this study was to probe effective surgical procedures and improve the outcome for patients with severe hepatic injury. A retrospective study of 348 patients with hepatic trauma seen in our institution during the past 12 years was carried out. Of these 348 patients, 259 (74.4%) underwent surgery. To manage severe liver trauma (American Association for the Surgery of Trauma grade III to grade V), procedures such as packing of the laceration with omentum, hepatectomy or direct control of bleeding vessels within the liver substance by means of the Pringle maneuver, selective hepatic artery ligation, retrohepatic caval repair with total hepatic vascular occlusion, and perihepatic packing were selected and combined based on the specific injury. In the 259 patients treated operatively, the survival rate was 86.9% (225/259); and 15 of 40 with retrohepatic venous injury (RHVI) were cured with the maximum blood transfusion of 60 units. In 42 patients treated by perihepatic packing, the bleeding was stopped in 20 of 25 (80%) with RHVI and in 14 of 17 (82%) without such injury ( p > 0.75). The percentage of failure of nonoperative management was 17.2% (17/99); and it was 46.7% (14/30) in patients with grade III-V injury. Death occurred in 3 (50%) of 6 failures of grade IV-V injury. The overall mortality rate was 11.8% (41/348), and 51% of the deaths were due to exsanguination. The results suggest that severe hepatic injuries, especially grade IV-V injuries, usually require surgical intervention; reasonable surgical procedures based on classification of liver trauma and combined application of techniques can increase the survival rate; and perihepatic packing is effective in dealing with RHVI. PMID- 12733003 TI - Insect cell culture for industrial production of recombinant proteins. AB - Insect cells used in conjunction with the baculovirus expression vector system (BEVS) are gaining ground rapidly as a platform for recombinant protein production. Insect cells present several comparative advantages to mammalian cells, such as ease of culture, higher tolerance to osmolality and by-product concentration and higher expression levels when infected with a recombinant baculovirus. Here we review some of the recent developments in protein expression by insect cells and their potential application in large-scale culture. Our current knowledge of insect cell metabolism is summarised and emphasis is placed on elements useful in the rational design of serum-free media. The culture of insect cells in the absence of serum is reaching maturity, and promising serum substitutes (hydrolysates, new growth and production-enhancing factors) are being evaluated. Proteolysis is a problem of the BEVS system due to its lytic nature, and can, therefore, be a critical issue in insect cell bioprocessing. Several cell- or baculovirus proteases are involved in degradation events during protein production by insect cells. Methods for proteolysis control, the optimal inhibitors and culture and storage conditions which affect proteolysis are discussed. Finally, engineering issues related to high-density culture (new bioreactor types, gas exchange, feeding strategies) are addressed in view of their relevance to large-scale culture. PMID- 12733004 TI - The constitutive AHSB4 promoter--a novel component of the Arxula adeninivorans based expression platform. AB - An Arxula adeninivorans-AHSB4 gene, encoding histone H4, was isolated and characterized. The gene includes a coding sequence of 363 bp disrupted by a 51-bp intron, similar to the situation in other fungal H4 genes. The identity of the gene was confirmed by the high degree of homology of the derived amino acid sequence with that of other H4 histones. The gene is strongly and constitutively expressed, maintaining this expression profile under salt-stress conditions. The AHSB4 promoter was tested for suitability in heterologous gene expression using genes encoding the intracellular green fluorescent protein and the secreted human serum albumin (HSA) for assessment. Plasmids incorporating respective expression cassettes were used to transform the host strain A. adeninivorans LS3, which forms budding cells at 30 degrees C, and strain 135, which forms mycelia under these conditions. Transformants of both types were found to harbor a single copy of the heterologous DNA. Strong constitutive expression was observed during culture in salt-containing and salt-free media, as expected from the expression profile of AHSB4. In 200-ml shake-flask cultures, maximal HSA levels of 20 mg l( 1) culture medium were achieved. This productivity could be increased to 50 mg l( 1 )in strains harboring two copies of the expression cassette. The AHSB4 promoter thus provides an attractive component for constitutive heterologous gene expression under salt-free and salt-stress conditions. PMID- 12733006 TI - Euroanalysis-12. The European Conference on Analytical Chemistry. September 8-13, 2003. Dortmund, Germany. PMID- 12733005 TI - Molecular evolution of V(H)9 germline genes isolated from DBA, BALB, 129 and C57BL mouse strains and sublines. AB - We have used the polymerase chain reaction (PCR) in an attempt to clone and sequence the exons and hitherto unavailable contiguous flanks of all members of the small V(H) 9 germline gene family from inbred mouse strains and sublines that have had a common ancestry within the last century, and to analyze the molecular evolution of these sequences. Fifteen genuine germline genes were isolated (designated V(H) 9.1 through V(H) 9.15) from strains and sublines of DBA, BALB, 129 and C57BL inbred mice. Of the 15 genuine isolates, nine are novel: seven sequences from DBA strains and sublines ( V(H) 9.3 to V(H) 9.9) and two sequences from C57BL strains ( V(H) 9.13 and V(H) 9.14). We have identified sequencing errors and PCR recombinant artefacts in previously published sequences. We detected no sequence divergence of individual genes shared by the strains and sublines studied. However, we isolated two genes from DBA strains and sublines, V(H) 9.1 and V(H) 9.3, that differ only by five nucleotides encoding three amino acid changes that are concentrated within a 33 nucleotide (11 codon) region. Of these 11 codons, eight encode a putative antigen binding site. There were no differences in the remaining 733 nucleotides sequenced (including both 5' and 3' flanking regions). Potential explanations for the generation of V(H) 9.1 and V(H) 9.3 are discussed. PMID- 12733007 TI - European analytical column No. 31(January 2003). PMID- 12733008 TI - PCR technology for screening and quantification of genetically modified organisms (GMOs). AB - Although PCR technology has obvious limitations, the potentially high degree of sensitivity and specificity explains why it has been the first choice of most analytical laboratories interested in detection of genetically modified (GM) organisms (GMOs) and derived materials. Because the products that laboratories receive for analysis are often processed and refined, the quality and quantity of target analyte (e.g. protein or DNA) frequently challenges the sensitivity of any detection method. Among the currently available methods, PCR methods are generally accepted as the most sensitive and reliable methods for detection of GM derived material in routine applications. The choice of target sequence motif is the single most important factor controlling the specificity of the PCR method. The target sequence is normally a part of the modified gene construct, for example a promoter, a terminator, a gene, or a junction between two of these elements. However, the elements may originate from wildtype organisms, they may be present in more than one GMO, and their copy number may also vary from one GMO to another. They may even be combined in a similar way in more than one GMO. Thus, the choice of method should fit the purpose. Recent developments include event-specific methods, particularly useful for identification and quantification of GM content. Thresholds for labelling are now in place in many countries including those in the European Union. The success of the labelling schemes is dependent upon the efficiency with which GM-derived material can be detected. We will present an overview of currently available PCR methods for screening and quantification of GM-derived DNA, and discuss their applicability and limitations. In addition, we will discuss some of the major challenges related to determination of the limits of detection (LOD) and quantification (LOQ), and to validation of methods. PMID- 12733009 TI - A combinatorial approach to obtain affinity media with binding properties towards the aflatoxins. AB - In our work we performed a combinatorial solid-phase synthesis in aqueous medium to prepare peptide libraries from which to select an amino acid sequence with binding properties towards aflatoxins. We used polystyrene beads, functionalised with carboxylic groups as solid support and eight amino acids as monomers. During the first step 64 different sequences of two amino acids were prepared by exploiting the principles of combinatorial chemistry; then the binding properties of all sequences towards aflatoxin B(1 )were checked. We determined binding constants towards aflatoxin B(1) and towards aflatoxins B(2), G(1) and G(2). Results were promising, so we prepared a new library by using the selected dipeptide as the starting solid phase. After selecting the best tetrapeptide sequences, we determined binding constants towards the quoted aflatoxins. We obtained binding constants ( K>10(4) M(-1)) similar to those shown by human serum albumin for similar compounds. Preliminary studies on an extraction column were promising for the development of an SPE system and for its application in food matrices. PMID- 12733010 TI - Molecular heterogeneity, detected by two electrophoretic micro procedures, of IgG in human cerebrospinal fluid (CSF). AB - Semi-automated electrophoretic procedures in the PhastSystem (Amersham Pharmacia Biotech) with micro polyacrylamide gels (PAGs) and SDS-PAG gradients were modified to analyze IgG in human cerebrospinal fluid (CSF) and matched serum samples with respect to the molecular IgG structure L-H-H-L. Isoelectric focusing (IEF) with specific immunofixation detected discrete IgG bands in CSF standing out against a polyclonal and monoclonal background pattern in CSF and serum; they were denoted oligoclonal bands (IgG OBs) (OB assay positive) indicating IgG synthesis in the central nervous system (CNS) of patients with subacute and chronic processes of inflammatory CNS disorders; assay was negative with identical (mirror) bands in CSF and serum for other CNS processes. IgG OBs were specified as lambda (kappa) IgG subfractions, precipitated with the anti-light (L) chains lambda (kappa) and anti-heavy (H) chain fragments (Fd, Fc, C(H)2) as well as with anti-F(ab')(2), and as duplex IgGs with kappa and lambda OBs at the same pI. With SDS-PAG gradient electrophoresis and specific immunofixation more than six IgG fractions were detected and classed according to apparent molecular weights of a S-sulfonated human IgG standard; they were characterized with the monospecific antibodies against the L and H chain fragments as 25, 50, 75, 100, 125 and 150 kD fractions containing combinations of L and H chains as well as mixtures of both L and H chain fragments of varying dimensions. Generally, this molecular IgG heterogeneity could not be connected with the IgG OB heterogeneity revealed by IEF; but single OBs in the strongly alkaline pH region of PAG may correspond to H fragments with basic pI. Nevertheless, evidence for the existence of both free L chains and the free H chain were revealed as specific OBs with IEF and with the anti-L and anti-H antibodies in the 25 kD and 50 kD fractions, respectively, of CSF samples of six patients with diverse CNS diseases. Further experiments are needed to elicit the origin of the molecular IgG heterogeneity during the immune response of subacute and chronic inflammatory processes in human CNS. PMID- 12733011 TI - Comparison of potassium ion preference of potassium-sensing oligonucleotides, PSO 1 and PSO-2, carrying the human and Oxytricha telomeric sequence, respectively. AB - Human [G(3)(TTAG(3))(3)] and Oxytricha [G(4)(T(4)G(4))(3)] telomere model oligonucleotides, PSO-1 and PSO-2, bearing two fluorophores, 6-carboxyfluorescein (6-FAM) and 6-carboxytetramethylrhodamine (6-TAMRA) at their 5'- and 3'-termini, respectively, were synthesized. Both of them can form an intramolecular antiparallel tetraplex upon addition of K(+), and an enhanced fluorescence resonance energy transfer (FRET) was observed. PSO-1 showed a 43,000 times higher selectivity for K(+) against Na(+). Fluorometric and circular dichroism spectrophotometric studies revealed that this system is useful for the evaluation of the interaction of different telomeric repeat oligonucleotide sequences with metal ions. PMID- 12733012 TI - Determination of antioxidant properties of aromatic herbs, olives and fresh fruit using an enzymatic sensor. AB - The aim was to experimentally evaluate the antioxidant capacity of different fresh aromatic herbs (field balm, marjoram, parsley, rosemary, sage, sweet basil), several varieties of olives from Central Italy ('Carboncello', 'Rosciolo', 'Olivastro', 'Coratello', 'Leccino', 'Frantoio') and several types of fresh fruit (apple, apricot, banana, cherry, fig, grape, medlar, melon, peach, pear, pineapple, plum, water melon, yellow plum) using a superoxide dismutase (SOD) biosensor developed by the present authors. Measurements were carried out by comparing the biosensor response to the concentration of superoxide radical produced in solution using a xanthine/xanthine oxidase system in the presence and absence of the antioxidant sample considered. Tests carried out on different samples of fruit and aromatic herbs showed that the homogenised samples had better antioxidant properties than the centrifuged ones (obtained by centrifuging the homogenate), which sometimes gave extremely low antioxidant capacity values. The reliability of the proposed method was checked by comparing the trend of some experimental results found using the SOD biosensor with those reported in the literature obtained using the classic (ORAC) method. The precision of this method of analysis was found to be good for samples of aromatic herbs (RSD% 0.16 mg/m(3) years) and inspirable dust (>2.9 mg/m(3) years) were two to four times more likely to report work-related wheeze and chest tightness than were unexposed subjects. Lower prevalence ratios for the same symptoms were seen with sulphur dioxide and BSF. Levels of lung function decreased slightly with exposure to oil mist, but not with cumulative exposure to other substances. CONCLUSIONS: This study suggests that the relevant causative agents for respiratory symptoms in aluminium smelters are fluoride and inspirable dust. PMID- 12733083 TI - Urinary protein excretion in humans exposed to arsenic and cadmium. AB - OBJECTIVE: In mice, the renal toxicity of arsenic (As) and cadmium (Cd) has been shown to be exacerbated by the simultaneous administration of both elements. To verify the existence of such an interaction in humans, cohorts slightly (Belgian) and moderately (Chinese) exposed to both elements were examined. METHODS: Biological indicators of exposure (Cd in urine and in blood; As in urine) and renal effect parameters (retinol binding protein (RBP); beta(2)-microglobulin (beta(2)M); albumin (ALB); N-acetyl-beta-glucosaminidase (NAG) in urine) were determined and their relationships studied by multiple regression analyses. RESULTS: Changes in renal effect parameters could be ascribed to Cd body burden. RBP and beta(2)M urinary concentration were influenced by exposure to As only in Chinese women, directly and in interaction with Cd exposure. CONCLUSION: A synergistic action of As on the tubular effects of Cd is observed in women moderately exposed to these elements and leads to RBP urinary excretion slightly above normal values. PMID- 12733081 TI - Working in permanent hypoxia for fire protection-impact on health. AB - OBJECTIVES: A new technique to prevent fires is continuous exchange of oxygen with nitrogen which leads to an oxygen concentration of between 15% and 13% in the ambient air. This paper reviews the effect of short-term, intermittent hypoxia on health and performance of people working in such atmospheres. METHODS: We reviewed the effect of ambient air hypoxia on human health in the literature using Medline, as well as reference lists of articles and handbooks. Articles were assessed from the perspective of working conditions in fire-protected rooms. RESULTS: Oxygen reduced to 15% and 13% in normobaric atmospheres is equivalent to the hypobaric atmospheres found at 2,700 and 3,850-m altitudes. When acutely exposed, a healthy person responds within minutes to hours with increased ventilation, stimulation of the sympathetic system, increased heart rate, increased pulmonary-circulation resistance, reduced plasma volume, and stimulation of erythropoesis. Acute mountain sickness occurs frequently at these oxygen partial pressures, but the full syndrome is rare if continuous exposure is limited to 6 h. Mood, cognitive, and psychomotor functions may be mildly impaired in these conditions, but data are inconclusive. Persons suffering from cardiac, pulmonary, or hematological diseases should consult a specialist in order for their individual risk to be assessed, and medical screening for any of these diseases is strongly recommended prior to exposure. CONCLUSION: Preliminary evidence suggests that working environments with low oxygen concentrations to a minimum of 13% and normal barometric pressure do not impose a health hazard, provided that precautions are observed, comprising medical examinations and limitation of exposure time. However, evidence is limited, particularly with regard to workers performing strenuous tasks or having various diseases. Therefore, close monitoring of the health problems of people working in low oxygen atmospheres is necessary. PMID- 12733085 TI - Proposal for single and mixture biological exposure limits for sevoflurane and nitrous oxide at low occupational exposure levels. AB - OBJECTIVES: Assessment of individual exposures to sevoflurane plus nitrous oxide (N(2)O) by biological monitoring of unmodified analytes in post-shift urine of exposed personnel. METHODS: Anaesthetics in urine and breathing area were monitored in 124 subjects in 11 operating theatres. Passive samplers were collected after 2.5-7 h of exposure, at the same time as post-shift urinary samples, to evaluate the individual time-weighted average (TWA) exposures to sevoflurane and N(2)O. A static headspace sampler coupled with a gas chromatograph mass spectrometer was used for analytical determinations (sensitivity sufficient to reveal biological/environmental exposures of 0.1 microg/l(urine) and 50 ppb for sevoflurane, and 1 microg/l(urine) and 80 ppb for N(2)O). RESULTS: Median (range) post-shift urinary and environmental values were 1.2 microg/l(urine) (0.1-5.0) and 0.4 ppm (0.05-3.0) for sevoflurane ( n=107) and 10.9 microg/l(urine) (0.5-74.9) and 8.6 ppm (0.2-123.4) for N(2)O ( n=121) (all low-exposure range). At log-log regression, urinary levels closely correlated with environmental data (sevoflurane, r(2)=0.7538; N(2)O, r(2)=0.8749). Biological equivalent limits (BELs) based on National Institute for Occupational Safety and Health (NIOSH) TWA exposure limits, calculated as means of regression slope and y-intercept, were 3.6 microg/l(urine) for sevoflurane (corresponding to 2 ppm) and 22.3 microg/l(urine) for N(2)O (corresponding to 25 ppm). Individual "mixture BELs", which we calculated by applying the American Conference of Governmental Industrial Hygienists (ACGIH) threshold limit value (TLV) mix formula to biomarker values and using the obtained NIOSH-based BELs as a reference, closely correlated with mixture TLVs (rho=0.816, Lin's concordance test). CONCLUSIONS. We propose urinary sevoflurane as a new, specific, internal dose biomarker for routine biological monitoring of personal exposures among operating-theatre personnel, and use of reliable "mixture BELs" to provide safer levels of internal exposure for workers exposed to mixtures of sevoflurane and N(2)O, and conceivably also to other mixtures of toxicants with possible additive effects. PMID- 12733084 TI - The effects of co-exposure to methyl ethyl ketone on the biological monitoring of occupational exposure to N,N-dimethylformamide. AB - OBJECTIVES: (1) To assess whether urinary N,N-dimethylformamide (U-DMF) is suitable as a biomarker when co-exposure to methyl ethyl ketone (MEK) exists, and to evaluate whether it is suitable as an exposure biomarker of DMF. (2) To examine whether the co-exposure to MEK affects the characteristics of U-NMF and U DMF. (3) To investigate if the difference in creatinine-adjusted and non-adjusted measurements of urinary biomarkers of DMF exposure is substantial. METHODS: Personal exposure monitoring of N,N-dimethylformamide (DMF) and MEK on 11 synthetic-leather workers was performed for 5 consecutive days. Daily post-shift urine for each individual was collected and was analyzed for urinary N methylformamide (U-NMF) and U-DMF levels on both non-adjusted and creatinine adjusted bases. RESULTS: Both U-NMF and U-DMF showed significant associations with airborne DMF. Positive and significant associations between U-NMF and U-DMF on either a non-adjusted basis or a creatinine-adjusted basis were found. Satisfactory linear associations ( P<0.01) between all kinds of urinary biomarkers and DMF exposure were found. The co-exposure to MEK exerted more effect on the relationship of airborne DMF to U-DMF than to U-NMF. CONCLUSIONS: U DMF is detectable when occupational DMF exposure is near or below the occupational exposure limit of 10 ppm. In view of the performance of sensitivity, specificity, and positive predictive value, U-NMF, in general, is superior to U DMF. However, on a par with other findings in this and previous studies, U-DMF might be considered as a complimentary biomarker of exposure to DMF in addition to U-NMF. No distinction between creatinine-adjustment or non-adjustment for urine specimens was found in the biological monitoring of DMF exposure. Further exploration of the influence of co-exposure to MEK at higher exposure is warranted. PMID- 12733086 TI - Sampling and analysis of carbon in diesel exhaust particulates--an international comparison. AB - OBJECTIVES: The European Co-ordination on Diesel Soot Exposure (ECDSE) working group organised a laboratory inter-comparison which integrated both the sampling and the analytical determination. The aim was to gain more information on the performance of the methods for the determination of diesel particulate matter at workplaces and on their comparability, and to also confirm that the requirements of the European standard EN 482 are fulfilled for this analytical procedure. METHODS: Sampling was carried out in a diesel aerosol test chamber where participants used their own sampling devices. Overall, ten sampling exercises at two different concentration levels were performed and both personal air sampling systems and stationary samplers were used. The analytical determination was performed according to the laboratory's own standard procedure. RESULTS: There was good agreement between the laboratories for the determination of organic carbon (OC), elemental carbon (EC) and total carbon (TC). Almost all the results when expressed as a coefficient of variation were in the range +/-30% of the overall means. The results also showed that the coefficient of variation for OC was approximately twice the coefficient of variation for EC. This is not too critical, because existing occupational exposure levels (OELs) are based on the measurement of EC. CONCLUSIONS: The inter-comparison showed that existing analytical procedures for the determination of diesel particulate matter at workplaces fulfil the requirements of European standard EN 482. Both personal air samplers and stationary samplers give comparable results. The parameters for the analytical determination are not critical within the range of parameter values presented in this article. Additionally, it may be concluded that the diesel aerosol test chamber used in this exercise is well suited for producing an atmosphere containing a constant and reproducible level of diesel particulate matter. PMID- 12733087 TI - Diagnostic management of orthostatic intolerance in the workplace. AB - OBJECTIVE: Orthostatic intolerance (OI) is a syndrome that is characterised by headache, concentration difficulties, palpitation of the heart, dizziness associated with postural tachycardia and plasma norepinephrine concentrations that are disproportionately high when the sufferer is in the upright posture. In contrast to other forms of orthostatic dysregulation - orthostatic hypotension (OH) and postural orthostatic tachycardia syndrome (POTS) - OI, hitherto, could be diagnosed only by a tilt table examination, with high expenditure. In this paper we examine the reliability and validity of a questionnaire as a screening instrument for OI. METHODS: We studied 138 young men (mean age 21.6 years) who were undergoing military service. After a medical check and filling in the questionnaire, the participants underwent a tilt table test including monitoring of blood pressure, heart rate and plasma catecholamines, in the supine position and during 30 min of standing. The questionnaire consisted of ten items registering presence and frequency of typical OI symptoms. RESULTS: Probands (104) showed normal tilt table test results. OI was diagnosed in 14 probands, OH in 6 and POTS in 14. The OI participants scored significantly higher in the questionnaire than the healthy subjects did: the mean score of the OI group was 22.6, the healthy participants had a mean score of 3.9. Participants with POTS had a mean score of 13.5 and subjects with OH had a mean score of 17.0. Reliability analysis showed a Cronbach's alpha of 0.888. Validity analysis showed that 93.5% of the probands with any kind of orthostatic dysregulation can be detected. CONCLUSIONS: We were able to establish a short questionnaire as a reliable and valid screening instrument for OI. Usage of this questionnaire can simplify enormously the diagnostic management of patients with suspected OI. PMID- 12733088 TI - Propylene glycol monomethyl ether occupational exposure (PGME). 4. Analysis of 2 methoxypropionic acid in urine. AB - OBJECTIVE: The two isomers propylene glycol monomethyl ether [PGME-alpha (1 methoxy-2-propanol, M2P) and PGME-beta (2-methoxy-1-propanol)] have different toxicities due to the different ways they are metabolised. The higher toxicity of PGME-beta has been attributed to the formation of 2-methoxypropionic acid (2-MPA) as a metabolite of primary alcohol. Six healthy male volunteers were exposed to PGME-alpha vapour (15, 50 and 95 ppm) with and without respiratory protection for 6 h, including a 30-min break. They were also exposed to PGME-alpha liquid (10% or 30% in water), via one hand, for 30 min or 1 h. Commercial products of M2P always contain a small quantity of the beta isomer, and GC analysis has shown that the product used for this human volunteer exposure contained approximately 0.3% of the beta isomer. The objective of this study was to determine the levels of 2-MPA in urine after these exposures to 99.7% PGME-alpha. METHOD: An analytical method developed by Laitinen [6] was used for the determination of 2 MPA in the urine of exposed volunteers. RESULTS: End exposure levels of 2-MPA were found to reach from 1.19 to 3.29 mg/l for inhalation and dermal exposure to PGME-alpha vapour and from under the detection limit to 2.10 mg/l for exposure of one hand in PGME-alpha liquid. 2-MPA concentrations in urine samples from a non exposed person or from a person exposed to PGME-alpha vapour at 15 ppm (inhalation and dermal exposure) and also from a person exposed to PGME-alpha vapour up to 95 ppm with respiratory protection (dermal-only exposure) all varied from under the detection limit to 0.30 mg/l and are then not significant. PMID- 12733089 TI - Heart-rate variability of carbon disulfide-poisoned subjects in Korea. AB - OBJECTIVES: Mass poisoning by carbon disulfide (CS(2)) occurred in a viscose rayon factory in Korea. Including 38 who had died, 830 employees were diagnosed as CS(2) poisoned. We evaluated the heart-rate variability (HRV) among CS(2) poisoned subjects, to discover whether CS(2) may affect HRV and whether its toxic effect persists after exposure has ceased. METHODS: The case group comprised 71 retired male workers with CS(2) poisoning. The control group comprised 127 public officials of the same age-range who had no history of organic-solvent exposure and cardiovascular diseases. Information on individual age, height, weight, cigarette smoking, alcohol drinking, regular exercise, medical and occupational history, and ECG recordings of the two groups were collected through medical examination and self-administered questionnaire. Time (maximum, average, minimum RR interval) and frequency domain measures [low (LF), high (HF), and total power spectrum (TPS)] of the two groups were analyzed. RESULTS: In univariate analysis, time domain measures of the two groups did not differ significantly, while frequency domain measures in the case group was significantly lower than in the control group except for HF power. In multivariate analysis, previous history of CS(2) poisoning was inversely related to all frequency domain measures and significantly affected LF power. CONCLUSIONS: This study suggests that CS(2) may cause heart-rate impairment and its toxic effect persists after exposure has ceased. Based on this study, HRV measurement can be considered as a useful tool to assess toxic effects of CS(2) both in current and retired workers. PMID- 12733090 TI - Three decades of pleural cancer and mesothelioma registration in Austria where asbestos cement was invented. AB - Recently, a new mesothelioma epidemic was predicted from observations made in Western Europe. From early observations in Austria the lower increase in cases of mesothelioma compared with neighbor countries had been related to different uses of asbestos. In order to test this hypothesis, incidence and mortality of pleural cancer [International Classification of Diseases (ICD)-8/9 163] were analyzed for three decades and supplemented by data from a cohort study in the factory that had been the largest consumer of asbestos imported to Austria and from all Austrian occupational diseases registered between 1990 and 2001. In men, mortality rates (based on 15 to 45 deaths/year) were lowest in 1980-1989, but similar in 1970-1979 and 1990-2001. No increase in younger-birth cohorts was detected. Incidence rates (based on 13 to 44 cases/year) increased (36%) non significantly ( P=0.14). In women, a significant decrease in mortality and incidence rates ( P<0.01) was observed from 1970. Rates from work-related mesothelioma (based on only 0-7 men and 0-4 women/year) must be interpreted with caution. In the cohort of 2,816 asbestos cement workers 26 pleural mesotheliomas were registered from 1990 through mid-1999. Six of these cases (three male and three female) had not been registered as an occupational disease, but all of these cases had been encoded under ICD 163 in mortality statistics. One female cohort member registered as having asbestosis according to the death certificate had died from mesothelioma according to the statistics of occupational diseases. We conclude that no epidemic of mesothelioma due to past asbestos exposure is to be expected in Austria. PMID- 12733091 TI - Sensitisation to occupational allergens in bakers' asthma and rhinitis: a case referent study. AB - OBJECTIVES: To study the importance of sensitisation to occupational allergens for the occurrence of asthma and rhinitis in bakers. METHODS: This is a nested clinical case-referent study of bakers based on a cohort of Swedish former bakery students. Cases were asthmatic ( n=25) or rhinitic bakers ( n=20). Randomly selected bakers ( n=44) were referents. All subjects underwent skin prick tests (SPTs) with common allergens, flours, fungal alpha-amylase and the storage mite L. destructor. Indices of airway inflammation were assessed in serum and the nose. RESULTS: Seven of the asthmatics and eight of the rhinitics reported onset of disease during bakery work. Flour SPTs were positive in 43% of the asthmatics or rhinitics vs 16% of referents. The corresponding figures for alpha-amylase were 29%, 25%, and 7%. The odds ratio, adjusted for atopy, for an SPT positive to flour or alpha-amylase for asthmatics with onset during bakery work was 5.8 (95% confidence interval 1.1-32), and 2.6 (0.4-16) for the corresponding rhinitics. The positive predictive value of sensitisation to flour or alpha-amylase in relation to a clinical diagnosis of asthma or rhinitis was 71%. Sensitisation to L. destructor was rare. The indices of airway inflammation were similar in cases and in referents. CONCLUSIONS: Bakers' asthma is associated with sensitisation to flour and/or alpha-amylase, atopy taken into account. A similar association was suggested in bakers' rhinitis. Indices of airway inflammation were of low predictive value for detecting bakers' asthma or rhinitis in this study. PMID- 12733092 TI - The Banfield Education Program. PMID- 12733093 TI - On-line continuing education at the College of Veterinary Medicine, Washington State University. PMID- 12733094 TI - Evaluating the stages of veterinary practitioner learning for continuing education needs assessment and program evaluation. AB - RATIONALE FOR THE STUDY: Recent behavioral change theory suggests that individuals go through stages of readiness for change. This theory has been applied to continuing medical education as a four stage theory of physician learning. The purpose of this project was to test a method that used the four stage learning theory to evaluate differences between continuing veterinary medical education (CVME) program attendees and non-attendees and to evaluate movement from one stage to another after a continuing education activity. METHODS: A survey using eight clinical scenarios was used to elucidate the stage of learning of dairy practitioner participants before and after a CVME course and of non-participating dairy practitioners. Differences in response rates before and after the course and between participants and non-participants were analyzed using chi-square contingency table analysis. RESULTS: Responses to six of the eight scenarios were different between participants and non-participants (p < 0.10). Attendees were more likely to report needing to update to solve the specific problems. Depending on the scenario, participants changed their responses after completing the continuing education course (range 31-81% change). CONCLUSIONS: The four stage theory of learning can be used for continuing education needs assessment, for understanding program participation, and for program evaluation. A continuing education activity can move practitioners from one stage of learning to another. PMID- 12733095 TI - Using clinical audits to identify practitioner learning needs. AB - Clinical audits are used in medicine to monitor practice performance, provide practice feedback, and identify learning needs. Although relatively new to veterinary practice, they hold promise as personal learning needs assessment tools. Continuing veterinary medical education providers are in a unique position to assist practitioners in the design, implementation, and evaluation of clinical audits. This paper discusses the types, the interpretation, and the potential uses of data from clinical audits in the veterinary setting and links audits to continuing veterinary medical education. PMID- 12733096 TI - The case for continuing education in veterinary colleges. PMID- 12733097 TI - Teaching the principles of health management to first year veterinary students. AB - A course called Health Management 1 was created as part of a new DVM curriculum at the Ontario Veterinary College. This full year course was designed to introduce students to basic concepts of health management, integrating the disciplines of epidemiology, ethology, and public health in the context of selected animal industries. The course was comprised of 60 lecture hours and four two-hour laboratories. A common definition of health management, incorporating five principles, was used throughout the course, in order to reinforce the concepts and to maintain continuity between lecture blocks. Unlike in the years prior to the introduction of the new curriculum, epidemiology was presented as a tool of health management rather than as a separate discipline. To supplement the lecture and laboratory material, a Web-based resource was created and the students were required to review the appropriate section prior to each lecture block. Small quizzes, consisting of 10 questions each within WebCT, were used to stimulate self-directed learning. Overall, the course was well received by the students. The Web resources combined with the WebCT quizzes proved to be an effective method of stimulating students to prepare for lecture. PMID- 12733098 TI - Importance of the human-animal bond for pre-veterinary, first-year, and fourth year veterinary students in relation to their career choice. AB - RATIONALE FOR THE STUDY: The Human-Animal Bond (HAB) is a construct that has received increased attention in the field of veterinary medicine. However, it remains unclear how important the HAB is to veterinary students and how it may be related to their career choice. METHODOLOGY: Questionnaires were administered to 146 veterinary students. A variety of variables was assessed, including sex, year of study, career choice, surgery track, and "farm" versus "city" upbringings. RESULTS AND CONCLUSIONS: Overall, students consider the HAB to be an important and valuable construct, one that was influential in their decision to become veterinarians. However, the HAB's importance seems to decrease as students progress in school. Also, students aspiring to food animal careers seem to attach less value to some aspects of the HAB. Females attached more importance to the role HAB plays in their lives than did males; those on "alternative" surgery track assigned more significance to the role of the HAB in veterinary medicine than did those on the "traditional" surgery track. Students also reported that they believe the HAB should be addressed in veterinary curricula. PMID- 12733100 TI - Veterinary student projects competing in the Hill's and the DHHS Secretary's Award Writing Competitions. PMID- 12733099 TI - Use of animation-enhanced video clips for teaching abnormal breathing patterns. AB - The ability to characterize disease of the respiratory tract accurately based on breathing pattern is helpful for the development of differential diagnoses and an efficient diagnostic plan and critical for the stabilization of patients in respiratory distress. Veterinary students do not have sufficient clinical experience to observe personally all types of respiratory diseases and their resultant abnormal breathing patterns. We developed a teaching tool, the animated breathing pattern videotape (ABV), to fill this gap. The ABV is a collection of video clips of small animal patients with normal and abnormal breathing patterns on a conventional videotape of approximately 20 minutes duration. Each video clip is shown for 20 to 40 seconds, followed by the same clip with superimposed animation of rib and diaphragm motion, followed by the initial clip again, without overlying animation. The ABV has since been used in teaching third-year veterinary students, interns, residents, practicing veterinarians, and veterinary technicians. Student evaluations and responses to questionnaires by interns, residents, practicing veterinarians, veterinary technicians, and peer reviewers have been uniformly positive. PMID- 12733101 TI - In retrospect: teaching is "dribbling and passing". AB - An introspective view of the author revealed that his philosophy, style, and methods of teaching were anchored in the broad fundamentals of considering, organizing, and implementing an educational plan. Key elements of these fundamentals included the art of teaching, time management and the commitment of time to students, experience and repetition, teaching methodology, communication and listening, relevancy of information, the laboratory teaching environment, and the responsibilities of students. A coherent approach to teaching was developed by blending these individual elements with a commitment to learning by all students. PMID- 12733102 TI - The application of the precautionary principle to the blood system: the Canadian blood system's vCJD donor deferral policy. AB - The precautionary principle is an influential concept that has been widely used in international treaties and declarations involving the protection of the environment. The principle is now being applied to the development of transfusion policy. In this article, we examine the application of the precautionary principle in the policy process leading to Canada's decision to defer donations from individuals who had traveled to the United Kingdom because of concerns over variant Creutzfeldt-Jakob disease. We found that, although the principle prominently influenced the decision-making process, problems existed with its interpretation. In particular, there was difficulty in balancing the risk prevented by applying a precautionary measure against the risk introduced by the same measure; in this case, the potential for shortages of blood. This dilemma is somewhat unique to the public health sector and will likely recur in future applications of the principle to transfusion policy. PMID- 12733103 TI - Islet transplantation, stem cells, and transfusion medicine. AB - Despite the widespread use of exogenous insulin, morbidity and mortality caused by type 1 diabetes mellitus (DM) continue to place a significant burden on society, both in terms of human suffering and cost. The transplantation of vascularized pancreas, usually performed concurrently with renal transplantation, can cure type 1 DM, as shown by results in more than 15000 such transplants over about 30 years. Transplantation of isolated pancreatic islets, instead of the whole organ, however, offers an attractive alternative that minimizes surgery and its complications. Although islet transplantation initially met with only modest success (only about 9% insulin independence at 1 year posttransplant), recent changes in patient selection criteria, number and treatment of islets transplanted, and better immunosuppressive regimens dramatically improved the results; spawning widespread enthusiasm for islet transplantation. Despite this promise, organ/islet availability remains an important limitation to this technology. A solution to the problem of limited materials for transplantation may be in the use of stem/progenitor cells. This article reviews the background of the current enthusiasm for pancreatic islet cell transplantation, highlights future research trends in the field, and suggests that the new islet-related cellular therapies belong within the domain of transfusion medicine. PMID- 12733104 TI - Review of the clinical practice literature on patient characteristics associated with perioperative allogeneic red blood cell transfusion. AB - There is evidence to suggest that there exists considerable variation in red blood cell (RBC) transfusion practices, especially in the surgical specialties. This is in large part related to difficulties in defining specific transfusion threshold criteria, given that there is no minimum acceptable hemoglobin threshold concentration and there is variability in assigning importance to patient factors. The purpose of this study is to identify patient-related factors that might be associated with the need for allogeneic RBC transfusion in surgical patients. We systematically identified, selected, and reviewed all observational or interventional studies describing patient-specific or related variables associated with the need for allogeneic RBC transfusion in the surgical patient population. We also evaluated the methodological characteristics of the individual studies. Sixty-two studies met our inclusion criteria and were analyzed for this review. Most of these studies were conducted in patients undergoing cardiac surgery (n = 30) and orthopedic surgery (n = 16). Decreased preoperative red cell reserve was most frequently associated with RBC transfusions, being identified as a significant variable in 46 studies. The other factors commonly associated with transfusion were advancing age (n = 28), female gender (n = 21), and small body size (n = 14). Only 2 studies attempted to prospectively validate a predictive model for RBC transfusion based on the variables identified. This systematic review shows that preoperative anemia, advancing age, female gender, and small body size are often associated with perioperative allogeneic RBC transfusion. However, the retrospective nature of most of the studies and the small sample sizes make it difficult to formulate a clinically useful prediction rules regarding allogeneic RBC transfusion. Ongoing research in designing large prospective cohort studies evaluating transfusion patterns are needed to further elucidate how patient characteristics impact the transfusion threshold. PMID- 12733105 TI - Risks associated with transfusion of cellular blood components in Canada. AB - We provide a comprehensive review of risks associated with allogeneic red blood cell and platelet transfusions in Canada. The review focuses on clinically symptomatic noninfectious transfusion risks (acute and delayed hemolytic, febrile nonhemolytic [FNHTR], allergic, volume overload, transfusion-related acute lung injury, graft-versus-host disease, and posttransfusion purpura) and the risk of clinically significant disease from transfusion-transmitted infections. Data sources include information from Canadian Blood Services, Hema-Quebec, Health Canada, and the Quebec Hemovigilance System as well as published information from research studies and international hemovigilance systems. We estimate that in 2000 the aggregate risk of potentially severe reactions (excluding FNHTR and minor allergic reactions) was 43.2 per 100000 red cell units (95% confidence interval [CI]: 38.7-48.1), affecting 337 recipients, and 125.7 per 100000 platelet pools of 5 units (95% CI: 100.8-154.9), affecting 88 recipients. The most frequent potentially severe outcomes for red cell transfusion were hemolytic reactions and volume overload and for platelet transfusion were major allergic reactions and bacterial contamination. The current risk of human immunodeficiency virus and hepatitis C virus transmission is approximately 1 in 4 million and 1 in 3 million units, respectively. These estimates are useful for decisions concerning transfusion therapy, the informed consent process, and for evaluating efficacy of interventions to reduce risk. PMID- 12733107 TI - Proceedings of the Boston workshop on cytogenetics and molecular genetics of hematologic malignancies, November 8 to 11, 2001: introduction. PMID- 12733108 TI - Genetic and molecular genetic studies in the diagnosis of myeloid diseases. PMID- 12733109 TI - Genetic and molecular genetic studies in the diagnosis of T and NK cell neoplasia. PMID- 12733110 TI - Genetic and molecular genetic studies in the diagnosis of T-cell malignancies. PMID- 12733111 TI - Genetic and molecular genetic studies in the diagnosis of B-cell lymphomas I: mantle cell lymphoma, follicular lymphoma, and Burkitt's lymphoma. PMID- 12733112 TI - Genetic and molecular genetic studies in the diagnosis of B-cell lymphomas: marginal zone lymphomas. PMID- 12733113 TI - Genetic and molecular genetic studies in the diagnosis of immune-related lymphoproliferative disorders. PMID- 12733114 TI - Genetic and molecular genetic studies in the diagnosis of atypical lymphoid hyperplasias versus lymphoma. PMID- 12733115 TI - Clinical applications of molecular genetic testing in hematologic malignancies: advantages and limitations. PMID- 12733116 TI - Clonality analysis for antigen receptor genes: preliminary results from the Biomed-2 concerted action PL 96-3936. PMID- 12733117 TI - Evaluation of HER-2/neu oncogene status in breast tumors on tissue microarrays. AB - The amplification and/or overexpression of the HER-2/neu oncogene and its encoded receptor protein are increasingly used for prognostication and prediction of therapeutic response to Herceptin in breast cancer. However, large-scale examination of archival tumor blocks by immunohistochemistry (IHC) or fluorescence in situ hybridization (FISH) is prohibitively laborious and technically challenging. The tissue microarray (TMA) technique enables hundreds of tumors to be studied simultaneously in a single experiment. To evaluate the HER-2/neu status of a selection of the breast tumors in our tumor bank, we constructed a TMA from 97 breast tumors, with a single 0.6-mm core per specimen. HER-2/neu gene amplification by FISH was found in 20 of the 87 interpretable cases (23%): in 14 of 14 IHC 3+ cases (100%), 5 of 8 IHC 2+ cases (62.5%) and 1 of 65 IHC 0/1+ cases (1.5%). Three of the 67 cases with no evidence of HER-2/neu gene amplification by FISH were moderately positive (2+) by IHC. A close relationship was observed between these 2 assays as applied to the TMA (95.4% concordance: 95% CI, -2.2% to 6.8%; P <0.0001), and both HER-2/neu gene amplification and protein overexpression were strongly associated with tumor grade, estrogen receptor status, and progesterone receptor status. Gene amplification was found in most of the tumors with high-level overexpression (IHC 3+) and not in the unequivocal IHC-negative cases. Complementary analysis by IHC and FISH are, however, recommended for tumors graded as 2+ by IHC, the group with the most result discrepancy. Hum Pathol 34:362-368. PMID- 12733118 TI - Benign cystic mesothelioma of the peritoneum: a clinicopathologic study of 17 cases and immunohistochemical analysis of estrogen and progesterone receptor status. AB - Benign cystic mesothelioma (BCM) is an uncommon lesion of the peritoneum occurring predominantly in women of reproductive age. Although most patients are managed by surgical resection, a reported high incidence of cyst recurrence has led to the use of hormonal therapy in isolated cases in an attempt to control cyst size and relieve local symptoms. To date, the estrogen receptor (ER) and progesterone receptor (PR) status of BCM has not been evaluated. Here we present our experience with 17 cases (13 women, 4 men) of BCM seen over a 19-year period, including an immunohistochemical analysis of ER and PR status in 14 cases. All lesions showed typical morphological features of BCM, and calretinin immunostaining was positive in 14 of 14 cases. Five patients experienced either 1 or 2 tumor recurrences, and no patients died of disease. One case was diffusely positive for ER only, 1 case was focally positive for PR only, and 1 case was focally positive for both ER and PR. Although immunohistochemical detection of female sex hormone receptors in BCM is uncommon, the focal presence of ER and/or PR in some lesions does provide weak biologic support for the use of hormonal manipulation as a therapeutic option. Hum Pathol 34:369-374. PMID- 12733119 TI - A novel microdissection and genotyping of follicular-derived thyroid tumors to predict aggressiveness. AB - Distinguishing thyroid follicular adenoma from minimally invasive or encapsulated angioinvasive carcinoma can be diagnostically challenging. In some cases, tumors are distorted, fragmented, or stripped of their capsule, and a definitive diagnosis becomes nearly impossible. In other cases, the foci of capsular and/or vascular invasion are subtle, thus making the diagnosis of carcinoma difficult. We developed a microdissection genotyping assay for assessing a panel of tumor suppressor genes for loss of heterozygosity mutations. The frequency of allelic loss (FAL) in follicular-derived neoplasms correlates with the histologic aggressiveness of the tumor. Furthermore, we calculated the amount of genetic heterogeneity within each tumor, as a second important measure of a tumor's ability for clonal expansion and a surrogate marker for its malignant potential. The follicular adenomas had a low FAL (average 9%) and low intratumoral heterogeneity (5% variability). The minimally invasive and encapsulated angioinvasive carcinomas had an intermediate FAL (average 30%) and intermediate intratumoral heterogeneity (10% variability). The widely invasive carcinomas had a high FAL (average 53%) and high intratumoral heterogeneity (24% variability). Although a larger retrospective study is needed to correlate genotyping studies with patient outcome and prognosis, our results indicate that performing a mutational genotyping assay can stratify tumors into the histologically well defined categories of adenomas, minimally invasive/angioinvasive carcinomas, and widely invasive follicular carcinomas. PMID- 12733120 TI - Distribution of monoclonal antiferritin antibody in Kaposi's sarcoma, Hodgkin's disease, and hepatocellular carcinoma. AB - The immunotherapeutic treatment of cancers using antibodies (naked or conjugated to a drug, toxin, or radionuclide) relies upon the preferential expression of a targeted antigen on the cancer cell compared to normal tissues. Polyclonal antiferritin antisera have shown selective distribution and therapeutic efficacy when radiolabeled in Hodgkin's disease and hepatoma. In this immunohistochemical study, we investigated the distribution of ferritin in tumors from 6 patients with Kaposi's sarcoma, 12 patients with Hodkgin's disease, and 9 patients with hepatoma, as well as in selected normal tissues. We found that the monoclonal antiferritin antibody binds primarily to histiocytes in samples from Kaposi's sarcoma and Hodgkin's disease. One hepatocellular carcinoma showed diffuse cytoplasmic staining with ferritin. Deposition of the monoclonal antibody was not detectable in the remaining hepatocellular carcinoma samples. PMID- 12733121 TI - p27 Kip1 expression is reduced in pancreatic carcinoma but has limited prognostic value. AB - p27(Kip1) is a cyclin-dependent kinase inhibitor whose loss in malignant tumors is associated with disease progression and an unfavorable clinical outcome. There is limited information about its expression in pancreatic carcinomas. In a previous Japanese study, p27(Kip1) loss was a powerful negative prognostic factor. In the present study, we assessed the expression of p27(Kip1) in resected pancreatic ductal adenocarcinomas from 46 European patients and in associated lymph node metastases from 13 patients, using a standard avidin-biotin peroxidase complex immunohistochemical method. We also analyzed the relationships among p27(Kip1) expression, pathologic features, and clinical outcome. The extent of p27(Kip1) expression (ie, the percentage of cells expressing p27(Kip1)) was lower in carcinomas than in nonneoplastic ductal epithelia. Carcinomas with <5% p27(Kip1) expression were more likely to be poorly or moderately differentiated than well differentiated (P =.022). p27(Kip1) expression did not correlate with patient gender, tumor maximum dimension, T classification, lymph node metastasis, or International Union Against Cancer stage. No significant difference was seen between the extent of p27(Kip1) expression in lymph node metastases and their corresponding primary tumors. Univariate survival analysis showed that an increased risk of death was associated with 2 established prognostic factors: tumor size >5 cm (P =.011) and incomplete surgical excision (P =.016). Trends toward worse survival for patients whose primary tumors had <4% p27(Kip1) expression (P =.060) and for patients whose lymph node metastases had <5% p27(Kip1) expression (P =.054) were seen, but multivariate analysis suggested that p27(Kip1) expression was not independently prognostic. The findings raise the possibility that abnormalities of p27(Kip1) play a role in the pathogenesis of pancreatic ductal adenocarcinoma. However, the extent of p27(Kip1) expression in lymph node metastases and primary tumors is similar. Also, reduced p27(Kip1) expression has limited prognostic value, at least in European patients. PMID- 12733122 TI - Evaluating the volume ratio of bone marrow affected by fibrosis: a parameter crucial for the prognostic significance of marrow fibrosis in chronic myeloid leukemia. AB - Marrow fibrosis (MF) is a complication of bone marrow neoplasms that usually impairs quality of life and shortens survival time. Proof and exact quantification of MF is not yet standardized, thus impeding the comparability of results and the evaluation of its prognostic impact. In this study on 360 bone marrow biopsy specimens from 135 patients with either chronic idiopathic myelofibrosis (CIMF) (evaluation group) or chronic myeloid leukemia (CML) (test group), marked differences were detected between six different approaches systematically compared with respect to proof and quantification of MF. A new volumetric approach quantifying the marrow volume affected by fibrosis turned out to be superior to all of the other morphometric methods considering practicability and specificity of results, and superior to a semiquantitative procedure considering sensitivity, precision, and reproducibility (P < 0.00005). The assessment of the marrow volume with fibrosis was the only feature of MF independently influencing the survival time of patients (test group with CML; multivariate analysis, P = 0.0008). We conclude that an approach estimating the marrow volume affected by fibrosis is the method of choice to exactly quantify and prove MF. The loss of marrow volume due to fibrosis appears to be crucial with respect to the prognostic significance of MF in CML. Hum Pathol 34:391-401. PMID- 12733123 TI - Cartilaginous metaplasia in calcified diabetic peripheral vascular disease: morphologic evidence of enchondral ossification. AB - The mechanism of arterial calcification is not clear. We examined histological sections of major arteries from lower extremities of two patients with longstanding type II (or non-insulin-dependent) diabetes mellitus, and found morphological evidence of cartilaginous metaplasia and ectopic ossification with associated severe medial arterial calcification and atherosclerosis. Hematoxylin and eosin, alcian blue, and toluidine blue stains were applied for the demonstration of cartilage cells and their specific matrix proteins, and immunohistochemical studies for type II collagen. To our knowledge, cartilaginous metaplasia has not previously been described in medium-sized human muscular arteries. This observation supports the hypothesis that active enchondral ossification may be a pathway leading to arterial calcification in diabetic obstructive peripheral vascular disease. PMID- 12733124 TI - Epstein-Barr virus-associated lymphoepithelioma-like carcinoma of the esophagus. AB - Epstein-Barr virus (EBV)-associated lymphoepithelioma-like carcinoma (LELC) rarely occurs in the esophagus. We report a case of such tumor arising in the esophagus of a 64-year-old Taiwanese woman. No tumors were detected outside the esophagus including nasopharynx by thorough clinical studies. She underwent subtotal esophagectomy. Light microscopy disclosed a poorly differentiated carcinoma morphologically reminiscent of nasopharyngeal undifferentiated carcinoma (lymphoepithelioma). Immunohistochemical stain for latent membrane protein-1 showed positivity on the tumor cells. The infiltrating lymphocytes were chiefly composed of CD8-positive cytotoxic T cells. EBV DNA was demonstrated by both nested polymerase chain reaction (PCR) in the main tumor and metastatic lymph node, and localization in the tumor cells by in situ PCR in situ hybridization (ISH). However, the result of EBV-encoded small RNA-1 ISH was negative. Our case suggests that LELC of the esophagus may be associated with EBV in the endemic area. Due to its distinct histological features, the association with EBV, and possible prognostic implication, LELC of the esophagus should be precisely diagnosed and discerned from the usual poorly differentiated carcinoma. PMID- 12733125 TI - Tamoxifen's apoptotic effect on the endometrium. PMID- 12733127 TI - Descriptive epidemiology of ovarian cancer in the United States, 1992-1997. PMID- 12733128 TI - Pathology and classification of ovarian tumors. PMID- 12733129 TI - Incidence of extraovarian primary cancers in the United States, 1992-1997. PMID- 12733130 TI - Stage at diagnosis of ovarian cancer in the United States, 1992-1997. PMID- 12733131 TI - Multiple primary cancers of the ovary in the United States, 1992-1997. PMID- 12733132 TI - Incidence of ovarian cancer by race and ethnicity in the United States, 1992 1997. PMID- 12733133 TI - Race/ethnic variations in ovarian cancer mortality in the United States, 1992 1997. PMID- 12733134 TI - Ovarian cancer in children and young adults in the United States, 1992-1997. PMID- 12733135 TI - Regional variations in ovarian cancer incidence in the United States, 1992-1997. PMID- 12733137 TI - Thyroid neoplasms after therapeutic radiation for malignancies during childhood or adolescence. AB - BACKGROUND: Recent data indicate that the risk of developing a thyroid neoplasm clearly is increased after high-dose, therapeutic radiation therapy during childhood. To better understand the time course, natural history, and histopathology of thyroid lesions that develop after high-dose irradiation, the authors undertook a retrospective study of all survivors of childhood and adolescent malignancies who were treated at Memorial Sloan-Kettering Cancer Center and who developed a clinically apparent thyroid neoplasm. METHODS: The authors searched the data base of the Department of Pediatrics, the hospital based tumor registry, and the hospital medical records database for patients with thyroid neoplasms. RESULTS: Thirty-three patients were identified who developed a thyroid neoplasm after therapeutic radiation. Primary diagnoses were Hodgkin disease (n = 18 patients), non-Hodgkin lymphoma (n = 10 patients), acute lymphoblastic leukemia (n = 2 patients), acute myeloid leukemia (n = 1 patient), Wilms tumor (n = 1 patient), and neuroblastoma (n = 1 patient). The median age at the time of diagnosis of the primary malignancy was 12.0 years (range, 3.7-18.3 years), the median radiation dose to the thyroid gland was 2400 centigrays (cGy; range, 1000-4200 cGy), and the median interval from the time of radiation therapy until the recognition of thyroid disease was 13.0 years (range, 6.2-30.1 years). Thirteen of 33 thyroid lesions (39%) were malignant (11 papillary carcinomas and 2 follicular carcinomas). Age at diagnosis, gender ratio, and time elapsed since initial treatment did not differ between patients with malignant and benign lesions, but the median radiation dose to the thyroid was lower in patients who had malignant disease compared with patients who had benign disease (2000 cGy vs. 2950 cGy; P = 0.03). Disease was confined to the neck in all patients who had malignant thyroid lesions; after a median follow-up of 6.5 years (range, 0.9-12 years), none of the patients developed progressive or recurrent disease. CONCLUSIONS: Data from this study suggest that a high proportion of clinically apparent thyroid neoplasms that develop after therapeutic radiation for a childhood malignancy are malignant. However, most of these thyroid malignancies do not appear to behave in an aggressive fashion. Because thyroid neoplasms may not become evident for decades after radiation therapy, all individuals who are at risk require life-long follow-up. PMID- 12733136 TI - The Gothenburg Breast Screening Trial. AB - BACKGROUND: Although there is evidence for a reduction in breast carcinoma mortality with mammographic screening, some doubts have been expressed, and there is still uncertainty regarding the age specific effects. METHODS: The authors report on a randomized, controlled trial of mammographic screening for breast carcinoma that was conducted among 51,611 women (21,650 women who were invited to a screening [the study group] and 29,961 women in a control group) ages 39-59 years in Gothenburg, Sweden. Among women in the study group, the screening interval was 18 months. The screening phase of the trial took place in 1982-1991, and follow-up for breast carcinoma mortality continued until December 31, 1996. Mortality from breast carcinoma was analyzed using a Poisson regression model. Overall and age specific effects of invitation to mammography screening on breast carcinoma mortality were calculated. Three mortality effects were estimated: the effect on deaths from breast tumors diagnosed during the screening phase of the trial, as assessed by an independent Endpoint Committee (the EPC evaluation model); the effect on deaths from breast carcinoma diagnosed during the screening phase of the trial, as determined by data from the National Cancer Registry and the National Cause of Death Register (the SCB evaluation model); and the effect on deaths from all breast carcinomas diagnosed up to December 31, 1996, as determined by the National Cancer Registry and the National Cause of Death Register (the SCB follow-up model). RESULTS: A nonsignificant, 21% reduction in the rate of mortality from breast carcinoma with invitation to screening was observed using the EPC evaluation model (relative risk [RR], 0.79; 95% confidence interval [95% CI], 0.58-1.08; P = 0.14); and a borderline significant, 23% rate reduction was observed using the SCB follow-up model (RR, 0.77; 95% CI, 0.60 1.00; P = 0.05). Age specific analyses yielded greater mortality rate reductions for the groups of women ages 39-44 years, 45-49 years, and 55-59 years, but there was no mortality rate reduction in the group of women ages 50-54 years. The effects of invitation to mammographic screening on the incidence of lymph node positive disease closely paralleled the effects of invitation on breast carcinoma mortality. The effect on breast carcinoma mortality was consistent with the effect on all-cause mortality, suggesting no bias in classification of cause of death. Breast carcinoma incidence in the study group was almost identical to the incidence in the control group after trial by screening had ended in the control group (RR, 0.98; 95% CI, 0.88-1.09; P = 0.7). CONCLUSIONS: The current results support the commonly observed 20-30% reduction in breast carcinoma mortality with invitation to screening. The impression that screening is less effective in women younger than 50 years may be an oversimplification. Age specific effects should be a target for further research. PMID- 12733138 TI - Simultaneous detection of apoptosis and proliferation in colorectal carcinoma by multiparameter flow cytometry allows separation of high and low-turnover tumors with distinct clinical outcome. AB - BACKGROUND: Dukes C colorectal carcinoma is treated with adjuvant chemotherapy. Adjuvant treatment is not standard for patients with Dukes B tumors, even though about 20% of patients within this tumor stage die of recurrent disease. The authors investigated whether a novel method of simultaneous detection of apoptosis and proliferation would improve the assessment of prognosis in colorectal carcinoma patients, with the ultimate goal of accurately identifying patients eligible for adjuvant therapy. METHODS: A multiparameter flow cytometric assay with heat pretreatment was performed on 278 paraffin-embedded colorectal adenocarcinomas. After immunochemical isolation of tumor cells, apoptosis and proliferation were assessed simultaneously by immunostaining for the M30 antibody and by quantitative DNA analysis, respectively. Patients were followed for more than 10 years. RESULTS: The mean values of apoptosis (apoptotic fraction [AF], i.e., the percentage of M30-positive cells) and proliferation (S-phase fraction [SPF]) were 11.1% and 13.1%, respectively. The AF and SPF values were correlated positively (P = 0.01) and both increased with advancing tumor stage (P = 0.02). Combining AF and SPF, patients with tumors with a high turnover (i.e., both AF and SPF values were greater than the mean) had an overall survival rate of 13%, whereas patients with low-turnover tumors (i.e., both AF and SPF values were less than the mean) had an overall survival rate of 89% (P < 0.001 by log rank test). Moreover, within Dukes B and C stages, patients with high-turnover tumors had a poorer survival than patients with low-turnover tumors (P < 0.001 for both stages). CONCLUSIONS: The simultaneous detection of apoptosis and proliferation in archival material allows separation of high and low-turnover colorectal adenocarcinomas and improves the assessment of prognosis. This technique could be used to stratify patients for adjuvant chemotherapy. PMID- 12733139 TI - Clinical significance of secreted protein acidic and rich in cystein in esophageal carcinoma and its relation to carcinoma progression. AB - BACKGROUND: Secreted protein acidic and rich in cystein (SPARC) is a small extramatrix-associated protein. Its production increases during angiogenesis and enhances matrix metalloproteinase-2 (MMP-2) expression. The goal of this study was to show the clinical relevance of SPARC and its relation to MMP-2 expression in esophageal carcinoma patients. METHODS: The authors investigated SPARC mRNA expression in 48 tissue samples of esophageal tumors characterized by MMP-2 mRNA expression in a Northern blot analysis. Western blot analysis and immunohistochemistry were also performed in esophageal carcinoma tissue samples. RESULTS: All 48 tissue specimens had high expression of SPARC mRNA. Quantitative evaluation showed that high SPARC mRNA was associated significantly with lymph node metastasis (P = 0.05) and poorer prognosis (P = 0.025). Expression of SPARC mRNA was associated significantly with MMP-2 mRNA expression (R = 0.65; P < 0.01). Both SPARC and MMP-2 were immunolocalized intensely in carcinoma and stromal cells, whereas normal esophageal mucosa and submucosa did not express SPARC. The 35-kilodalton cleaved SPARC was detected in esophageal carcinoma tissue specimens by Western blot analysis and it was associated with MMP-2 mRNA expression. CONCLUSIONS: In terms of clinical significance, SPARC accumulation may reflect a functional correlation with MMP-2 and the associated expression could play a key role in the progression of esophageal carcinoma. PMID- 12733141 TI - Transitional cell carcinoma of the urinary bladder with regional lymph node involvement treated by cystectomy: clinicopathologic features associated with outcome. AB - BACKGROUND: Patients with transitional cell carcinoma (TCC) of the urinary bladder metastatic to regional lymph nodes (LN) typically have a poor prognosis. However, some patients are cured by radical cystectomy alone. The goal of this study was to identify predictors of survival in this cohort. METHODS: The authors identified 154 patients with TCC metastatic to regional LNs treated by cystectomy between 1970 and 1998. Clinical characteristics collected included age, gender, and preoperative computed tomographic or magnetic resonance image scan findings, as well as neoadjuvant and adjuvant therapy. Pathologic features evaluated included multifocality, size, pathologic stage, grade, and margin status of the primary tumor, as well as the number, location, and bilaterality of the positive LNs. Capsular penetration, greatest linear extent, and surface area of the largest metastatic LN deposit were also recorded. The Kaplan-Meier method was used to evaluate survival rates. Cox proportional hazards models were used to identify predictors of outcome. RESULTS: The mean follow-up was 4.5 years (range, 0.1-13.9 years). In a multivariate setting, only adjuvant chemotherapy and the number of positive LNs were associated significantly with death from TCC. Patients treated adjuvantly with chemotherapy were 2.1 times less likely to die of their disease (P = 0.005). Each increase in one positive LN increased the risk of death from TCC by 20% (P < 0.001). Recursive partitioning indicated that the optimal cutoff point to predict death from TCC was five or more positive LNs. CONCLUSIONS: Adjuvant chemotherapy and the number of positive LNs were associated significantly with death from TCC. PMID- 12733140 TI - Evaluation of an automated immunochemical fecal occult blood test for colorectal neoplasia detection in a Chinese population. AB - BACKGROUND: Most commercial fecal occult blood tests (FOBT) used for colorectal carcinoma screening of Western populations are guaiac-based, manually developed, subjective, and sensitive to dietary components. Preliminary studies demonstrated the unsuitability of these tests for screening a Chinese population. The goal of the current study was to evaluate the performance characteristics of a human hemoglobin-specific automated immunochemical FOBT, the Magstream 1000/Hem SP (Fujirebio, Inc., Tokyo, Japan), in a Chinese population referred for colonoscopy. METHODS: Two hundred fifty consecutive patients who were referred for colonoscopy and met the study inclusion criteria provided samples for the immunochemical FOBT (without dietary restrictions) from two successive stool specimens. Tests were developed with an automated instrument that had an adjustable sensitivity threshold. The sensitivity, specificity, and positive predictive value for detecting colorectal adenomas and carcinomas were calculated according to the manufacturer's instructions over a range of sensitivity levels. RESULTS: At the optimal threshold level, the sensitivity, specificity, and positive predictive value for detection of significant colorectal neoplasia (adenomas >or= 1.0 cm and carcinomas) were 62%, 93%, and 44%, respectively. The test was easy to use, and results did not depend on operator experience. CONCLUSIONS: The automated immunochemical FOBT used in the current study was a robust, convenient, and useful tool for colorectal carcinoma screening in the study population. PMID- 12733142 TI - Subsequent primary malignancies after endometrial carcinoma and ovarian carcinoma. AB - BACKGROUND: Population-based data on subsequent neoplasms after women are diagnosed with endometrial and ovarian carcinomas are limited, particularly regarding specific histologic tumor types. METHODS: The nationwide Swedish Family Cancer Database of 10.2 million individuals, which includes 19,128 invasive endometrial carcinomas and 19,440 ovarian carcinomas, was used to calculate standardized incidence ratios (SIRs) and 95% confidence intervals (95% CIs) for second primary carcinomas. SIRs were calculated for specific follow-up periods. Data on histopathologic types also were used. RESULTS: An excess of subsequent malignancies after women were diagnosed with endometrial carcinoma was noted at 11 sites. The highest SIRs were recorded for synchronous or metasynchronous ovarian carcinomas (SIR, 55.77; 95% CI, 48.82-63.43) and carcinomas of the small intestines (SIR, 14.71; 95% CI, 4.64-34.59). Primary ovarian carcinoma was followed by an increased risk of developing endometrial carcinoma, and the risks of developing many other malignancies also were increased after women were diagnosed with endometrial carcinoma, including intestinal malignancies, renal cell carcinoma, bladder carcinoma, squamous cell skin carcinoma, connective tissue malignancies, and leukemia. When ovarian endometrioid histology was diagnosed synchronously with primary endometrial carcinoma, the SIR was 140; when endometrial carcinoma was the subsequent neoplasm, the SIR was 87. A small familial component was found in the cooccurrence of endometrial carcinoma and ovarian carcinoma. CONCLUSIONS: The current data show a strong clustering of endometrial carcinomas and ovarian carcinomas, particularly involving tumors of endometrioid morphology. The patterns of second neoplasms also suggest that hereditary nonpolyposis colorectal carcinoma may contribute to the association between endometrial and ovarian malignancies. Increased risks for connective tissue tumors and leukemia may signal a response to treatment, and an increased risk for squamous cell skin carcinoma may signal a depressed immune function. PMID- 12733143 TI - A review of the cytokine network in multiple myeloma: diagnostic, prognostic, and therapeutic implications. AB - Because many studies have focused on growth factors in multiple myeloma, the study of the cytokine network appears to be useful for this purpose. Interleukin 6 (IL-6) and IL-2 with their soluble receptors (IL-3, IL-4, IL-10, and IL-11) have been examined. Plasma cells may produce IL-6 by an autocrine mechanism whereas a paracrine mechanism is believed to be involved in the production of IL 6 by bone marrow stromal cells through an interaction between adhesion molecules present on myeloma plasma cells and their respective receptors that are present on bone marrow stromal cells. In addition, control over production of IL-6 may be exerted by other ILs such as IL-1beta and IL-10. Among target cells, the growth of normal and myeloma plasma cells is supported by IL-6, which also induces the differentiation of myeloma plasmablastic cells into mature plasma cells. This last action also is shared by IL-3, IL-4, and, most likely, IL-8. Evaluation of the serum level of IL-6, C reactive protein, soluble IL-6 receptor (sIL-6R), and soluble IL-2 receptor (sIL-2R), together with the activity exerted by IL-3 and IL 4 on some cellular subsets, may constitute an additional element in the differential diagnosis of borderline cases. However, the concomitant evaluation of all immunologic parameters could be more useful than the value of a single IL. Serum levels of IL-6, sIL-6R, sIL-2R, and the expression of membrane-bound IL-2 receptors, both on bone marrow plasma cells and on peripheral blood mononuclear cells, are correlated with disease activity and disease stage. In addition, IL-6 and sIL-6R serum levels are believed to be correlated with the duration of disease-free survival because a high serum level at the time of diagnosis is believed to be correlated with a short duration of survival. However, some laboratory parameters may express the same prognostic value as high beta(2) microglobulin and lactate dehydrogenase (LDH) serum levels together with a high plasma cell labeling index are correlated with disease activity. Furthermore, if the evaluation is performed at the time of diagnosis, high values of these parameters are correlated with a short disease-free survival. A correlation between laboratory parameters and the serum level of several cytokines was demonstrated. Hence, the real advantage of the prognostic evaluation of cytokines is reserved for patients who do not exhibit uniform results with regard to beta(2) microglobulin and LDH serum levels, or, better, for borderline cases. With regard to the differential diagnosis, all immunologic parameters should be evaluated concomitantly rather than separately to confer a real prognostic value to results. Furthermore, a particular relation was found between a high sIL-6R serum level and a poor response to chemotherapy, therefore suggesting the possibility of identifying in advance a subset of patients with a high risk of treatment failure, as has already been demonstrated in other hematologic malignancies.Finally, the majority of studies indicate that interferons are used mainly in the immunotherapy for multiple myeloma, whereas many clinical trials should still be required for the evaluation of the effectiveness of anti-I-L6 antibodies or antiidiotypic vaccines in reference to the eligible patients for these particular therapies. PMID- 12733144 TI - Avascular necrosis in long-term survivors after allogeneic or autologous stem cell transplantation: a single center experience and a review. AB - BACKGROUND: The most debilitating skeletal complication of stem cell transplantation (SCT) is avascular necrosis (AVN). METHODS: Two hundred seven consecutive patients were evaluated prospectively for AVN. They survived disease free for more than 180 days after autologous or allogeneic SCT for hematologic malignancies. The diagnosis of AVN in suspicious cases was confirmed by magnetic resonance imaging. Possible correlations with treatments, bone mineral density (BMD), graft versus host disease (GVHD), and in vitro growth of fibroblast progenitors were investigated. Bone mineral density was evaluated by dual-energy X-ray absorptiometry in 100 transplanted patients, and the in vitro growth of fibroblast progenitors was monitored by a fibroblast colony-forming unit (CFU-F) assay in 30 patients after allogeneic SCT. RESULTS: Twelve patients developed AVN 3-114 months (median, 26 months) following SCT: 10 (10%) after allogeneic SCT and 2 (1.9%) after autologous SCT (P = 0.04). Twenty-five joints were affected by AVN. All patients had femoral head involvement, which was managed with hip replacement in six of them. All but one patient who developed AVN after allogeneic SCT suffered from chronic GVHD (cGVHD). Avascular necrosis occurred 1 4 months after exacerbation or progression of cGVHD. Cumulative dose of steroids was similar in both SCT groups (including steroids given pretransplant for the basic disease), whereas treatment duration was significantly longer in the allogeneic SCT group. Avascular necrosis was related to the decreased number of bone marrow CFU-F colonies in vitro, but not to BMD values. CONCLUSIONS: Avascular necrosis is a skeletal complication that occurs more often after allogeneic than after autologous SCT. Occurrence of AVN symptoms after clinical follow-up of cGVHD suggests that cGVHD requiring long-term steroid therapy is one of the main risk factors for AVN. Avascular necrosis may be facilitated by a severe deficit in the repopulating capacity of bone marrow stromal stem cells after SCT. PMID- 12733145 TI - Primary gastrointestinal lymphoma in Japan: a clinicopathologic analysis of 455 patients with special reference to its time trends. AB - BACKGROUND: An optimal treatment modality for patients with primary gastrointestinal lymphoma has not yet been established. This study aimed to elucidate the clinicopathologic features of this disease and the influence of therapeutic modalities on the prognosis in Japanese patients METHODS: The clinicopathologic features of 455 patients with primary gastrointestinal lymphoma were investigated retrospectively regarding treatment modalities and time trends. RESULTS: This study comprised 342 patients (75%) with gastric lymphoma, 96 patients (22%) with intestinal lymphoma, and 17 patients (4%) with both gastric and intestinal lymphoma. Two hundred thirty-one (51%) patients were classified as having low-grade B-cell lymphoma including 200 marginal zone lymphoma of mucosa associated lymphoid tissue (MALT) type, 185 (41%) patients were classified as having high-grade B-cell lymphoma including 76 diffuse large cell lymphoma plus MALT lymphoma, and 39 (9%) patients were classified as having T-cell lymphoma. The frequency of nonsurgical treatment, including Helicobacter pylori eradication, chemotherapy, and radiation, increased during the latest decade. Patients who received nonsurgical treatment showed a better overall survival than those treated by surgery, but event-free survival did not differ between two groups. Cox multivariate analysis revealed that early stage, younger age, gastric localization, B-cell phenotype, and absence of B symptoms were independent prognostic factors for better overall and event-free survivals. Mucosa-associated lymphoid tissue-derived lymphoma was also an independent prognostic factor for event-free survival, but not for overall survival. CONCLUSIONS: Nonsurgical treatment may be an optimal therapeutic modality for patients with primary gastrointestinal lymphoma. PMID- 12733146 TI - Identification of the antigens predominantly reacted with serum from patients with hepatocellular carcinoma. AB - BACKGROUND: To identify antigens specifically recognized by the immune surveillance system in patients with hepatocellular carcinoma (HCC), the authors examined two complementary DNA (cDNA) libraries of moderately differentiated HCC by serologic analysis of recombinant cDNA expression libraries (SEREX). METHODS: The libraries were screened with autologous patients' sera, and sequences of the reacted clones were determined. To study the immunoreactivity of the antigens, sera from 20 patients with HCC, from 20 healthy volunteers, and from 16 patients with chronic viral hepatitis were examined. RESULTS: Twenty-seven antigens were identified. They included SART1, p57Kip2, ROCK-1, gamma-catenin, and heat shock proteins, which are classified as tumor-associated genes. Three of 27 antigens Tat-binding protein-1 (TBP-1), beta4 integrin-binding protein (p27[BBP]), and ribosomal protein L30 (rpL30)-were reacted predominantly with sera from patients with HCC (55% of patients, 45% of patients, and 20% of patients, respectively). Patients in the control group had no antibodies against these three antigens. Seventy percent of patients with HCC had the antibody against at least one of these antigens. CONCLUSIONS: Disease specific humoral immune response against TBP 1, p27(BBP), and rpL30 was induced in patients with HCC, and the antibodies against these antigens also may be used as tumor markers. PMID- 12733147 TI - Weekly, high-dose paclitaxel in advanced lung carcinoma: a phase II study with pharmacokinetics by the Cancer and Leukemia Group B. AB - BACKGROUND: The Cancer and Leukemia Group B conducted a Phase II trial to evaluate the efficacy, toxicity, and pharmacokinetics of paclitaxel administered at a maximum dose density for patients with chemotherapy-naive, advanced-stage non-small cell lung carcinoma (NSCLC). METHODS: Patients with Stage IIIB/IV or recurrent NSCLC, a performance status (PS) score of 0-1, and no history of chemotherapy exposure were eligible. Paclitaxel, 150 mg/m(2), was administered over 3 hours during Weeks 1-6 of an 8-week cycle. Doses were modified for ANC < 1500/microL or for >or= Grade 2 neuropathy on the day of therapy. Treatment continued until toxicity or disease progression. Pharmacokinetics were assessed at Weeks 1, 3, and 5 of Cycle 1. RESULTS: Thirty-eight patients (median age, 64 years; range, 31-81 years) were treated. There were 21 males (PS = 0 for 17). Eleven patients had received previous radiation, 2 had brain metastases, 25 had adenocarcinoma, 23 had Stage IV disease, 6 had StageIIIB disease, and 9 had recurrent disease. Grade 3-4 granulocytopenia occurred in 39% of patients. There were no deaths due to toxicity. Grade 2 or 3 neuropathy occurred in 29% and 24% of patients, respectively. Ten (27%) patients had Grade 3 hyperglycemia (glucose concentration > 250 mg/dL). There were 16 partial responses (42%; 95% confidence interval [CI], 26-59%). The median survival period was 12.3 months (95% CI, 7.9 19.6%), and the 1-year and 2-year survival rates were 52% (95% CI, 39-71%) and 26% (95% CI, 15-45%), respectively. Paclitaxel pharmacokinetics were consistent with published values and clearance was not induced. Older age and hyperglycemia were associated with greater neurotoxicity. CONCLUSIONS: Paclitaxel at 150 mg/m(2) per week x 6 every 8 weeks can be administered safely in the cooperative group setting. These Phase II data are comparable to those associated with combination therapy. The weekly dose-dense schedule may be more active than conventional schedules. PMID- 12733148 TI - Prognostic implications of neuroendocrine differentiation and hormone production in patients with Stage I nonsmall cell lung carcinoma. AB - BACKGROUND: Approximately 10-20% of nonsmall cell lung carcinomas (NSCLC) show neuroendocrine (NE) differentiation, as evaluated by panendocrine markers or ultrastructural evidence of dense-core secretory granules. However, little is known regarding the prevalence and clinical implications of NE differentiation in patients with Stage I NSCLC. METHODS: The authors analyzed 220 consecutive patients with Stage I NSCLC (pT1-T2N0M0) among 2100 patients with primary lung carcinoma who underwent surgical treatment between 1987 and 1993. Using light microscopy and immunohistochemical staining for synaptophysin, chromogranin A, and respiratory tract-related hormones, 28 NSCLC specimens with NE differentiation (NSCLC-ND) and 11 large cell neuroendocrine carcinoma (LCNEC) specimens were identified. RESULTS: The 28 NSCLC-ND specimens included 15 adenocarcinomas and 13 squamous cell carcinomas. Neoplastic cells with NE features never exceeded 20% in NSCLC-ND specimens, whereas neoplastic cells amounted to 20-90% in LCNEC specimens. NSCLC-ND specimens with > 5% NE differentiated tumor cells showed increased Ki-67 labeling index (P = 0.007) and invasive phenotype, as evaluated by fascin immunoreactivity (P = 0.021). Patients with adenocarcinoma, but not with squamous cell carcinoma, who had > 5% NE differentiated cells had a worse clinical course compared with patients who had ordinary NSCLC, with reduced overall survival (P = 0.017) and disease free survival (P = 0.049). In multivariate analysis, NE differentiation > 5% neoplastic cells in patients with adenocarcinoma independently predicted a poorer prognosis (hazard ratio, 2.61; 95% confidence interval, 0.99-6.85). Hormone production was restricted to chromogranin positive NSCLC-ND but did not affect prognosis. CONCLUSIONS: Stage I adenocarcinomas with >or= 5% NE tumor cells are clinically aggressive tumors, similar to LCNEC. Hormone production identifies a more fully developed neuroendocrine phenotype but is not relevant to prognosis. The identification of NE-differentiated cells in patients with NSCLC may have clinical relevance. PMID- 12733149 TI - Alternating chemotherapy with etoposide plus cisplatin and topotecan plus paclitaxel in patients with untreated, extensive-stage small cell lung carcinoma: a phase II trial of the North Central Cancer Treatment Group. AB - BACKGROUND: The objective of this study was to test the response rate and toxicity of alternating chemotherapy in previously untreated patients with extensive-stage small cell lung carcinoma (SCLC). METHODS: Patients with histologically proven, extensive-stage SCLC, with a performance status of 0-2, and who had received no prior chemotherapy were eligible. The design was a two stage, Phase II, multicenter trial. Treatment consisted of alternating chemotherapy every 3 weeks with etoposide (100 mg/m(2) on Days 1-3) and cisplatin (30 mg/m(2) on Days 1-3) on Cycles 1, 3, 5 and with topotecan (1 mg/m(2) on Days 1-5) and paclitaxel (200 mg/m(2) on Day 5) on Cycles 2, 4, and 6. Filgrastim support was given with Cycles 2, 4, 6. RESULTS: Forty-four patients were eligible and evaluable. The primary toxicity was myelosuppression. The median absolute neutrophil count was 300/microL with 70% Grade 4 neutropenia. The median platelet count was 58,000/microL with 23% Grade 4 thrombocytopenia. Grade 4 nonhematologic toxicities occurred in 16% of patients. Overall toxicities were not different between the two regimens. There were no treatment-related deaths. Complete or partial responses occurred in 34 patients (77%). The median time to progression was 6.9 months, with a median survival of 10.5 months and with 1-year and 2-year survival rates of 37% and 12%, respectively. CONCLUSIONS: The regimen of alternating chemotherapy was associated with substantial myelosuppression and resulted in a high response rate and good overall survival. The results were similar to those reported in prior trials and did not suggest any improvement in therapy for patients with SCLC. PMID- 12733150 TI - Sensitive detection of small cell lung carcinoma cells by reverse transcriptase polymerase chain reaction for prepro-gastrin-releasing peptide mRNA. AB - BACKGROUND: Gastrin-releasing peptide (GRP) is an autocrine growth factor in patients with small cell lung carcinoma (SCLC). The authors developed a reverse transcriptase-polymerase chain reaction (RT-PCR) assay for the detection of SCLC cells in the peripheral blood and the pleural effusion using preproGRP mRNA as a target. METHODS: The current study was conducted to determine the utility of preproGRP-specific nested RT-PCR on the peripheral blood, bone marrow, and pleural effusion samples from 32 patients with SCLC, 39 patients with non-small cell lung carcinoma (NSCLC), 28 patients with nonmalignant pulmonary disease, and 20 healthy volunteers. The internal primers were designed to amplify a 244-base pair PCR product, a sequence encompassing exon 1 and exon 2 by the nested RT-PCR assay. RESULTS: Amplification of the preproGRP message was detected in SCLC cell lines (LU165, SBC1, SBC2, and SBC3) but not in other NSCLC cell lines (A549, ABC1, EBC1, and Oka-1). The SCLC cells (LU165) were detected in dilutions of tumor cells of up to 10(-7) in hematopoietic cells from healthy donors. The preproGRP mRNA was detected in 16 of 32 (50%) blood samples, 2 of 11 (18%) marrow samples, and in all 6 (100%) pleural effusion samples. Blood samples gave positive results in 11 of 19 (58%) patients with extensive disease compared with 5 of 13 (38%) patients with limited disease. In contrast, only 1 blood sample (2.6%) from a patient with lung adenocarcinoma gave a positive result among patients with NSCLC. No other samples of blood, bone marrow, and pleural effusion from patients with NSCLC and none of the blood samples from patients with nonmalignant diseases and healthy volunteers were positive. CONCLUSIONS: The current RT-PCR approach may be a sensitive and specific assay to detect SCLC cells in circulating blood as well as in pleural effusions from SCLC patients. PMID- 12733151 TI - Developmental model relating white matter volume to neurocognitive deficits in pediatric brain tumor survivors. AB - BACKGROUND: The primary objective of this study was to test the hypothesis that, among survivors of pediatric brain tumors, the association between reduced volumes of normal-appearing white matter (NAWM) and intellectual/academic achievement deficits can be explained by patient problems with memory and attention. METHODS: Quantitative tissue volumes from magnetic resonance imaging scans and neurocognitive assessments were obtained for 40 long-term survivors of pediatric brain tumors. They were treated with radiotherapy (RT) with or without chemotherapy 2.6-15.3 years earlier (median, 5.7 years) at an age of 1.7-14.8 years (median, 6.5 years). Neurocognitive assessments included standardized tests of intellect (intelligence quotient [IQ]), attention, memory, and academic achievement. RESULTS: Analyses revealed significant impairments in patients' neurocognitive test performance on all measures. After statistically controlling for age at RT and time from RT, significant associations were found between NAWM volumes and both attentional abilities and IQ, and between attentional abilities and IQ. Subsequent analyses supported the hypothesis that attentional abilities, but not memory, could explain a significant amount of the relationship between NAWM and IQ. The final developmental model predicting academic achievement based on NAWM, attentional abilities, and IQ explained approximately 60% of the variance in reading and spelling and almost 80% of the variance in math. CONCLUSIONS: The authors demonstrated that the primary consequence of reduced NAWM among pediatric patients treated for brain tumors was decreased attentional abilities, leading to declining IQ and academic achievement. PMID- 12733152 TI - Malignancy in giant cell tumor of bone. AB - BACKGROUND: The term malignant giant cell tumor embraces multiple entities and therefore can be confusing. The goals of the current study were to define the clinicopathologic and histologic features of malignancy in giant cell tumors and to clarify the terminology. METHODS: The authors reviewed all cases from the Rizzoli Institute (Bologna, Italy) of primary (PMGCT) and secondary (SMGCT) malignancy in giant cell tumors. PMGCT is a high-grade sarcoma that arises side by side with benign giant cell tumors. SMGCT is a high-grade sarcoma that occurs at the sites of previously treated giant cell tumors of bone. RESULTS: The authors report 5 PMGCTs and 12 SMGCTs; half of the SMGCTs were postradiation sarcomas. Patient age ranged from 20 to 68 years (median, 62 years) for PMGCT and from 30 to 77 years (median, 40 years) for SMGCT. The average latent period between diagnosis of giant cell tumor and diagnosis of SMGCT was 9 years (range, 3-15 years) for patients with postradiation SMGCT and 19 years (range, 7-28 years) for patients with SMGCT resulting from spontaneous transformation. The histologic classification of high-grade sarcomas in the PMGCT group was osteosarcoma in four cases and malignant fibrous histiocytoma in one case. In the SMGCT group, the histologic classification was osteosarcoma in nine cases, fibrosarcoma in two cases, and malignant fibrous histiocytoma in one case. The outcomes associated with all malignancies in giant cell tumors were poor, with the worst outcome associated with postradiation SMGCT. CONCLUSIONS: Malignancies in giant cell tumors of bone always are high-grade sarcomas with a poor prognosis. These lesions must be distinguished from benign giant cell tumors of bone. SMGCT usually is easy to diagnose upon malignant clinicoradiographic presentation. In contrast, PMGCT often mimics giant cell tumors both clinically and radiographically. In addition, upon histologic examination, PMGCT shows areas of conventional giant cell tumor, which can lead to difficulties in making the correct diagnosis. PMID- 12733153 TI - Prognostic factors for patients with localized soft-tissue sarcoma treated with conservation surgery and radiation therapy: an analysis of 1225 patients. AB - BACKGROUND: Prognostic factors for patients with soft-tissue sarcoma who are treated with conservative surgery and radiation are documented poorly. METHODS: The clinicopathologic features and disease outcome for 1225 patients with localized sarcoma who were treated with conservative surgery and radiation were reviewed retrospectively. Actuarial univariate and multivariate statistical methods were used to determine significant prognostic factors for local control, metastatic recurrence, and disease specific survival. RESULTS: The median follow up of surviving patients was 9.5 years. The respective local control rates at 5 years, 10 years, and 15 years were 83%, 80%, and 79%. Factors predictive of local recurrence were positive or uncertain resection margins; tumors located in the head and neck and the deep trunk; presentation with local recurrence; patient age > 64 years; malignant fibrous histiocytoma, neurogenic sarcoma. or epithelioid sarcoma histopathology; tumor measuring > 10 cm in greatest dimension; and high pathologic grade. Freedom from metastasis at 5 years, 10 years, and 15 years was 71%, 68%, and 66%, respectively. Factors that were predictive of metastatic recurrence were high tumor grade; large tumor size (> 5 cm); and leiomyosarcoma, rhabdomyosarcoma, synovial sarcoma, or epithelioid sarcoma. The respective disease specific survival rates at 5 years, 10 years, and 15 years were 73%, 68%, and 65%. Adverse factors for disease specific survival were high tumor grade; large tumor size (> 5 cm); tumors located in the head and neck and deep trunk; rhabdomyosarcoma, epithelioid sarcoma, or clear cell sarcoma; patient age > 64 years; and positive or uncertain resection margins. CONCLUSIONS: Soft-tissue sarcoma comprises a heterogeneous group of diseases. Prognostic factors for local recurrence, metastatic recurrence, lymph node recurrence, disease free survival, and disease specific survival are different, and optimal treatment strategies need to take this complexity into account. PMID- 12733154 TI - Surgical margins and reresection in the management of patients with soft tissue sarcoma using conservative surgery and radiation therapy. AB - BACKGROUND: Patients with localized soft tissue sarcoma (STS) who present to specialist centers after undergoing apparent macroscopic total resection often have a significant incidence of residual tumor and may benefit from reresection of the tumor bed. The potential benefits of such reresection have not been documented adequately. METHODS: The clinicopathologic features and disease outcome for 666 consecutive patients with localized STS who presented after undergoing apparent macroscopic total tumor resection were analyzed to elucidate the relative merits of reresection. Actuarial univariate and multivariate methods were used to compare disease outcome of patients who presented with positive or uncertain microscopic resection margins according to whether they underwent reresection. All patients received adjuvant radiation therapy. RESULTS: Two hundred and ninety-five patients underwent reresection of their tumor bed, and residual tumor was found in 136 patients (46%), including macroscopic tumor in 73 patients (28%). Final resection margins among patients who underwent reresection were negative in 257 patients (87%), positive in 35 patients (12%), and uncertain in 3 patients (1%). Patients who did not undergo reresection had final margins that were negative in 117 patients (32%), positive in 47 patients (13%), and uncertain in 207 patients (56%). Local control rates at 5 years, 10 years, and 15 years for patients who underwent reresection were 85%, 85%, and 82%, respectively; for patients who did not undergo reresection, the respective local control rates were 78%, 73%, and 73% (P = 0.03). Reresection remained a significant determinant of local control when other prognostic factors were incorporated into a multivariate proportional hazards regression analysis. A similar beneficial effect of reresection was found for metastasis free survival and disease specific survival. CONCLUSIONS: Patients with localized STS who were referred to a specialist center after undergoing apparent macroscopic total resection of their tumor had a high incidence of residual tumor in their tumor bed and benefited from undergoing reresection, even if radiation was administered routinely. PMID- 12733155 TI - Education, employment, insurance, and marital status among 694 survivors of pediatric lower extremity bone tumors: a report from the childhood cancer survivor study. AB - BACKGROUND: With increasing numbers of childhood cancer survivors, direct sequelae of cancer therapy and psychosocial outcomes are becoming more important. The authors described psychosocial outcomes (education, employment, health insurance, and marriage) for survivors of pediatric lower extremity bone tumors. METHODS: The long-term follow-up study of the Childhood Cancer Survivor Study is a multiinstitutional cohort study comprising 14,054 individuals who have survived for 5 or more years after treatment for cancer diagnosed during childhood or adolescence. Baseline demographic and medical information were obtained. Six hundred ninety-four survivors had osteosarcoma or Ewing sarcoma of the lower extremity or pelvis and were classified by amputation status and by age at diagnosis. The median age at diagnosis was 14 years old with a median of 16 years of follow up since diagnosis. Demographic characteristics were used to analyze the rates of psychosocial outcomes. RESULTS: Amputation status and age at diagnosis did not significantly influence any of the measured psychosocial outcomes. Education was a significant positive predictor of employment, having health insurance, and being currently in their first marriage. Male gender predicted ever being employed and female gender predicted having health insurance and marriage. When compared with siblings, amputees had significant deficits in education, employment, and health insurance. CONCLUSIONS: Overall, no differences between amputees and nonamputees were found. However, gender and education play a prominent role. When compared with siblings, amputees in this cohort may benefit from additional supports. PMID- 12733156 TI - Physical activity and breast cancer risk among Asian-American women in Los Angeles: a case-control study. AB - BACKGROUND: To the authors' knowledge, there have been few studies published to date regarding physical activity patterns and breast cancer risk in Asian and Asian-American women. METHODS: The authors conducted a population-based case control study of 501 Asian-American women with incident breast cancer and a control group of 594 Asian-American women in Los Angeles County to evaluate the role of lifetime physical activity on breast cancer risk. Information concerning lifetime recreational physical activity (i.e., type of activity, duration [years], and frequency [average hours per week]) and occupational physical activity was obtained using a structured questionnaire that was administered in person. RESULTS: Increasing years and levels (average metabolic equivalent [MET] hours per week) of lifetime recreational activity were associated with a significantly reduced risk of breast cancer after adjusting for demographic factors, migration history, and menstrual and reproductive factors. Compared with women who had no lifetime recreational physical activity, 3-6 MET hours per week, > 6-12 MET hours per week, and > 12 MET hours per week of activity were associated with significantly reduced risk, with odds ratios (and 95% confidence intervals) of 0.91 (0.55-1.49), 0.65 (0.39-1.10), 0.53 (0.31-0.90), and 0.47 (0.28-0.80), respectively (P value for trend < 0.001). The risk of breast cancer was associated inversely with occupational physical activity, although the result was not statistically significant. CONCLUSIONS: The findings of the current study provide further support for the finding that physical activity has a protective role in breast cancer. PMID- 12733157 TI - Common community respiratory viruses in patients with cancer: more than just "common colds". AB - Community respiratory viruses long have been recognized as primary respiratory pathogens among infants and young children. More recently, it has become clear that these viruses cause a considerable disease burden throughout life. The consequences of repeated infections are most evident in elderly and immunocompromised persons. Even in otherwise healthy persons, reinfections often require medical attention but generally are undiagnosed and unrecognized. These reinfections may spread from healthy persons to those at highest risk. Control requires a multifaceted approach combining vaccination, chemoprophylaxis, and aggressive early antiviral treatment of high-risk individuals, as well as education of all populations affected by these viruses. PMID- 12733158 TI - Second neoplasms after megavoltage radiation for pediatric tumors. AB - BACKGROUND: Because ionizing radiation is a known carcinogen, diligent long-term follow-up in children exposed to therapeutic radiation is required. The authors updated an earlier study on the nature and risk of second neoplasms (SN) in patients treated with megavoltage radiotherapy as children. METHODS: The authors followed 446 children who were treated for a primary malignancy with megavoltage radiotherapy between January 16, 1954 and December 31, 1980. These children survived a minimum of 5 years. The nature and incidence of SNs were evaluated in this population. Patients with bilateral retinoblastoma or neurofibromatosis were excluded from the study due to their large genetically based predisposition for developing an SN. RESULTS: The Kaplan-Meier actuarial survival rate was 80% (95% confidence interval [CI] 74-85%) at 30 years for all patients. Thirty-seven (8.3%) patients developed SNs, most of which occurred within the original radiation treatment field, 3.8-31.8 years (median, 15.5 years) after radiotherapy. The cumulative risk of developing an SN was 13% (95% CI 9-19%) at 30 years and the standardized incidence ratio for the development of any SN was 5.2 (95% CI 3.4-7.6%). The most common SNs were breast carcinoma (n = 8), skin carcinoma (five basal cell carcinomas, two malignant melanomas, and one dermatosarcoma), and meningiomas (n = 6). All eight breast carcinomas occurred after the treatment of childhood Hodgkin disease. Of the 37 patients with SNs, 12 died of either the SN (n = 10) or of recurrent disease (n = 2). Risk factors associated with developing a SN included initial diagnosis of Hodgkin disease (P = 0.0003), female gender (P = 0.008), and an initial diagnosis of acute lymphoblastic leukemia (P = 0.02). CONCLUSIONS: Patients in the radiation-treated cohort experienced increased mortality, were at an increased risk of developing an SN, and should undergo increased medical surveillance as adults. The cumulative probability of developing an SN has increased substantially at 30 years, largely due to an increase in follow-up time. In addition, the cumulative probability curve does not show evidence of plateau after increased duration of follow-up. Finally, the emergence of secondary breast carcinoma in the current study was not noted in the previous analysis. PMID- 12733159 TI - Rhabdomyosarcoma in infants younger than one year old: a report from the Italian Cooperative Group. AB - BACKGROUND: The management of rhabdomyosarcoma (RMS) in patients age < 1 year is particularly problematic and requires a tailored therapeutic approach. We report on the Italian Cooperative Group's 20-year study of 50 children with RMS who were age < 1 year at diagnosis. METHODS: Patients were treated using multimodality therapeutic approaches that were based on three consecutive protocols. Chemotherapy was administered to all patients, with dosages calculated according to body weight; calculated doses subsequently were reduced by 33-50%. Radiotherapy was administered to 10 patients. RESULTS: With a median follow-up of 76 months, 5-year event-free survival and overall survival rates were 42.3% and 61.7%, respectively. Local recurrence was the major reason for treatment failure. In particular, the local recurrence rate was high in patients who warranted radiotherapy but received none due to their age. Completeness of surgery and nodal involvement were the most significant prognostic factors. After a suitable reduction in dosage was made, acute toxicity was no different from what has been observed in older children. The most relevant toxic event was cardiotoxic death in a newborn (n = 1). CONCLUSIONS: The current study confirmed that the outcome for infants with RMS is less satisfactory than for older children and that infants with RMS require more careful monitoring and specific treatment guidelines. The absence of local control is the major cause of treatment failure; aggressive conservative surgery should be encouraged, but more radiotherapy may be advisable in selected cases. Intensive chemotherapy is essential; a 33% dose reduction may ensure adequate tolerance. In addition, patients age < 3 months should not receive anthracyclines. PMID- 12733160 TI - Development of a brief screening interview for adjustment disorders and major depression in patients with cancer. AB - BACKGROUND: Adjustment disorders and major depression are common psychiatric disorders in patients with cancer and have a serious impact on quality of life. The problem in clinical oncology settings is underrecognition of these disorders; as a result, screening is recommended to detect them. The goal of the current study was to develop a new, brief screening tool for adjustment disorders and major depression and to compare its performance with that of existing screening methods. METHODS: Patients with cancer completed the newly developed One-Question Interview (a 1-item, structured interview); the Distress Thermometer (a 1-item, self-report questionnaire), which previously was developed as a brief screening tool; and the Hospital Anxiety and Depression Scale (HADS; a 14-item, self-report questionnaire). Psychiatric diagnoses of adjustment disorders and major depression were made by psychiatrists and were based on criteria set forth by the Diagnostic and Statistical Manual of Mental Disorders, 4th edition. RESULTS: Two hundred seventy-five patients participated in the study. Scores on both the One Question Interview and the Distress Thermometer were significantly correlated with HADS score (One-Question Interview: r = -0.66, P < 0.01; Distress Thermometer: r = 0.71, P < 0.01). At the optimal cutoff points, the sensitivity and specificity for detection of adjustment disorders and major depression were 80% and 61%, respectively, for the One-Question Interview; 84% and 61%, respectively, for the Distress Thermometer; and 92% and 57%, respectively, for the HADS. CONCLUSIONS: The results of the current study suggested that the One Question Interview was a valid tool for use in screening patients with cancer for adjustment disorders and major depression. Its performance was inferior to that of the HADS but comparable to that of the Distress Thermometer. The One-Question Interview may be suitable for widespread use in routine screening. PMID- 12733161 TI - Palliative care--a beginning. PMID- 12733162 TI - Prevalence of dry eye in University Malaya Medical Centre. AB - Two hundred outpatients (115 females and 85 males) attending the University Malaya Medical Center (UMMC) eye clinic, aged 20 years and above and without any ocular surface disorder were recruited for this study. Their tear film status was determined subjectively by their symptoms and quantitatively by the cotton thread test, Schirmer's test, marginal tear film meniscus assessment, fluorescein corneal staining and tear break-up time. Dry eye was considered present if at least one symptom was experienced often or always, within the past 3 months. Dry eye was also considered present if one of these tests was positive; Schirmer's test < 5 mm, Phenol red thread (PRT) test < 10 mm and tear film breakup time (BUT) < 8 seconds. The prevalence of dry eye in this sample population as defined by presence of symptoms and an abnormal test result is 14.5%. Presence of dry eye as detected by clinical testing is higher in the Chinese race (p < 0.01), in the group 40-59 years (p = 0.024). There is no difference between females and males. A lower BUT score was more strongly associated with presence of dry eye symptoms (p = 0.02). Elderly patients have a lower BUT and Schirmer's score. There is lack of agreement between PRT and Schirmer's test, although both are measures of tear quantity. PMID- 12733163 TI - Orthognathic surgery in the University of Malaya. AB - This is the first review on orthognathic surgery in Malaysia. The records of a total of 84 patients seen between 1977 and 1999 in the Department of Oral and Maxillofacial Surgery of the Faculty of Dentistry, University of Malaya were analysed. Skeletal III deformity formed 85% of the sample with a female dominance of 2 to 1. The patients' age ranged from 17 to 36 years, with a mean of 25.3 years. The common surgical techniques used were combined bilateral sagittal split and Le Fort I osteotomy. The predominant ethnic group was Chinese (n = 58, 69%); followed by Malay (n = 14, 17%) and Indian (n = 12, 14%). PMID- 12733164 TI - Endonasal endoscopic dacryocystorhinostomy. AB - A review of endoscopic dacryocystorhinostomy (EDCR) performed in University Hospital from 1998 till early 2001 were carried out. They were reviewed for indication, functional results, complications and other associated factors. Eleven patients (6 female and 5 male) were reviewed. All had unilateral nasolacrimal duct (NLD) obstruction (6 right, 5 left). Their ages range from 6 to 73 years. All 11 had persistent epiphora while 4 had concurrent recurrent dacryocystitis with purulent discharge. Two patients had history of lacrimal sac abscess that needed drainage. Ten patients had the procedure performed successfully. Functional results were excellent in 8 patients (80%) while two patients still had occasional epiphora although markedly reduced. Complications include synachae (1 patient), premature dislodgement of Jones tube (1 patient) and impacted tube in a child. Our experience with EDCR concludes it to be an easy, efficient treatment for nasolacrimal duct obstruction with minimal complications. PMID- 12733166 TI - A review of burns patients admitted to the Burns Unit of Hospital Universiti Kebangsaan Malaysia. AB - This is a retrospective review of 110 patients admitted to the Burns Units between October 1999 and November 2001. The aim was to determine the burns pattern of patients admitted to hospital UKM. There was an increasing trend for patients admitted. Female to male ratio was 1:2. Children consisted 34% of the total admission. Children had significant higher number of scald burns as compare to adult (p < 0.01). Domestic burns were consist of 75% overall admission. Mean percentage of TBSA (total body surface area) burns was 19%. Thirty percent of patients sustained more than 20% of TBSA. Sixty percent of patients had scald burns. Ninety percents of patients with second degree burns that were treated with biologic membrane dressing or split skin graft. Mean duration of hospital stay was 10 days. Over 70% of patients were discharged within 15 days. Overall mortality rate was 6.3%. The patients who died had significantly larger area of burns of more than 20% TBSA (p < 0.05) and a higher incidence of inhalation injury (p < 0.02). Hence, this study suggests a need for better preventive measures by the authority to prevent burns related accident and the expansion of the service provided by the Burns Unit. PMID- 12733165 TI - Palliative care: experience with 156 cases in a general hospital. AB - The concept of Palliative Care is still quite new in Malaysia and there is a need to promote the awareness of its importance for patients with incurable and advanced illnesses, not only to the public but also to the nursing and medical professionals. These patients especially the poor ones very often have no one to turn to; they cannot afford to seek treatment from the private hospitals, they are turned away from acute General Hospitals and are told that there is nothing more to be done because their illnesses are no longer curable, they cannot pay for GPs to come to their homes, and there is difficulty in purchasing opiate drugs for pain relief. This is a retrospective observational study of the palliative care services we try to provide to those few patients referred to us. This study showed that out of the total of 156 patients, majority were Chinese, peak age between 50-59 years, referrals were mainly from the community and the Obstetrics + Gynaecology department, patients were referred rather late, 60% of patients died at home, most common malignancies being those of the breast, colorectal and cervix, common sites of metastases being the lungs, liver, bones, peritoneum and local infiltration, 87% of patients experienced pain, about 40% of patients were not fully aware of both diagnosis and prognosis, common drugs used being opiate analgesics followed by dexamethasone, H2 antagonist, antiemetics and NSAID. PMID- 12733167 TI - Vascular trauma in Penang and Kuala Lumpur Hospitals. AB - OBJECTIVES: The nature of vascular trauma varies greatly between continents and across time. The aim of this study was to prospectively analyse the demographics, pathology, management and clinical outcomes of vascular injuries in two urban Malaysian hospitals and review of international literature on vascular trauma. From this information, preliminary management and preventive implications will be described. METHODS: Eighty-four consecutive cases of trauma requiring vascular surgery were prospectively analysed over three years at Hospital Kuala Lumpur and Hospital Pulau Pinang, Malaysia. Extensive patient demographic and injury data, including the mechanism of injury, associated injuries, angiographic findings, operative details and post-operative complications, were systematically gathered. RESULTS: Most vascular injuries were incurred by males (76/84), with 37% (28/76) of them aged between 21 and 30 years. Malays were most frequently injured (n = 36) followed by Chinese and Indians. Road traffic accidents (n = 49) substantially outnumbered all other causes of injury. Lower limb injuries (n = 57) occurred more than twice as often as upper limb injuries (n = 27). Complete arterial transections (n = 43) and intimal injuries (n = 27) were more common than arterial lacerations (n = 10) and pseudoaneurysms (n = 4). The most frequently damaged vessels were the popliteal/tibioperoneal trunk (n = 33). All patients received urgent Doppler ultrasound assessment and, where possible, ankle brachial systolic index measurement. Of all patients, 40 received an angiogram, haemodynamic instability making this investigation impractical in others. Primary arterial repair was the most frequently employed surgical procedure (n = 54) followed by autogenous reverse long saphenous vein (LSV) interposition graft (n = 14), embolectomy (n = 5) and PTFE interposition graft (n = 3). The most common post-operative complication was wound infection (n = 11). Amputation, as a last resort, was required in 13 cases following either primary or autogenous reverse LSV repair complicated by sepsis or critical ischaemia. CONCLUSIONS: Vascular trauma, especially in conjunction with severe soft tissue, nerve or orthopaedic injury carries colossal physical, psychological, financial and social costs. Associated nerve and venous injury portended poor outcome in this study. Whilst orthopaedic trauma was a common association, the concurrence of occult vascular trauma and soft tissue injury without fracture emphasises the crucial importance of thorough and rapid clinical vascular assessment, investigation and surgical intervention. Fasciotomy, especially for the lower limb, is important for the prevention of compartment syndrome and its, limb-threatening sequelae. Primary preventive road safety promotion and interventions, with attention to high-risk groups (young males and motorcyclists), is urgently required. PMID- 12733168 TI - Stigma in mental illness: attitudes of medical students towards mental illness. AB - BACKGROUND: Negative attitudes towards people with mental illness can be attributed to stigma. The objective of this study was to determine the attitudes of medical students towards mental illness by comparing those who have had contact with mental patients and those who have not. This study also assesses to what extent knowledge about mental illness can affect the students' attitude. METHODS: A vignette and two dependent measures (social distance scale and dangerousness scale) were used to assess the attitudes of medical students towards mental illness. They comprised of 108 first year and 85 final year medical students in University of Malaya, Kuala Lumpur. The first year students didn't have any prior psychiatric training. RESULTS: The final year students who had knowledge and contact (undergone 8 weeks of clinical psychiatric training) were less stigmatizing toward mentally ill patients. There were no significant differences in the attitudes towards mentally ill patient among the first year students (no knowledge) regardless they had previous contact or not. CONCLUSION: Knowledge seems to have the effect in inculcating greater tolerance of mental illness. Contact by itself is not sufficient for attitude changes. PMID- 12733169 TI - The performance properties of the International Index of Erectile Function (IIEF 15) in assessing erectile dysfunction in patients with lower urinary tract symptoms. AB - To validate the International Index of Erectile Function (IIEF-15) in Malaysian population. Reliability and internal consistency were evaluated using the test retest method and Cronbach's alpha. Sensitivity to change was expressed as the effect size index. Internal consistency was excellent (Cronbach's alpha value = 0.75 to 0.90) Test-retest correlation coefficient and intraclass correlation coefficient were highly significant (ICC = 0.75 and above) and a high degree of sensitivity and specificity was observed. The IIEF-15 is suitable, reliable, valid and sensitive to clinical change in the Malaysian population. PMID- 12733171 TI - The ethnic characteristics and prevalence of diabetes mellitus, hypertension and hyperlipidaemia in patients who underwent coronary artery bypass grafting in Hospital Universiti Kebangsaan Malaysia. AB - A retrospective study was done on 302 patients who had undergone coronary artery bypass grafting (CABG) in Hospital Universiti Kebangsaan Malaysia--46.0% were Chinese, 40.1% were Malays and 11.6% were Indians. Overall and irrespective of race and sex, the prevalence of diabetes mellitus, hypertension and hyperlipidaemia was 45.7%, 78.8% and 89.1% respectively indicating that hyperlipidaemia was the most prevalent risk factor amongst this cohort. The Indians had the highest prevalence of the three risk factors. The Chinese and the Malays most frequently presented with the combination of hypertension and hyperlipidaemia. PMID- 12733170 TI - Introducing routine external cephalic version for the management of the malpresenting fetus near term. AB - BACKGROUND: The aim of this study was to assess the efficacy and safety of external cephalic version (ECV) when its use was introduced in the routine management of breech presentation and transverse lie after 36 weeks by obstetricians with limited prior experience with the procedure. The influence of various factors on the outcome of ECV was also studied. METHODS: Retrospective study of 44 consecutive cases of ECV which were analysed with respect to outcome, parity, type of breech, placental site and birth weight. RESULTS: ECV was successful in 45% of women, 80% of women with successful ECV delivered vaginally while 10% underwent spontaneous reversion to a non-cephalic presentation. In contrast, only 15% of women with failed ECV delivered vaginally. Parity, type of breech presentation and placental location did not significantly affect the outcome of ECV although there was a trend towards better success rate of ECV with multiparity, flexed breech presentation, transverse lie and posteriorly-located placentae. The mean birth weight of fetuses of women with successful ECV was significantly heavier than those of women who failed ECV (p < 0.001). No significant fetal or maternal morbidity occurred as a result of ECV in this study. CONCLUSION: ECV is a safe and effective procedure that is useful in the management of breech presentation and transverse lie near term. The lack of prior experience with the procedure does not appear to influence the success rate or morbidity. PMID- 12733172 TI - Meningioma--the Sarawak General Hospital experience. AB - The aim of this study was to determine the factors for the large size of intracranial meningiomas at the time of presentation to the Sarawak General Hospital. The data was collected prospectively from 1/3/2000 to 28/2/2001. During this period a total of 57 cases of intracranial tumours were operated upon. Twenty of these cases (35%) were meningioma, making meningioma the most common intracranial tumour operated in the Neurosurgery service here with one to two cases operated per month. Headache was the most common symptom. The average duration of symptoms before the diagnosis was made was twenty-five months, the longest being fifteen years. The patients needed an average of seven visits prior to the diagnosis. PMID- 12733173 TI - Parental asthma knowledge. AB - Asthma knowledge an important components of asthma education. OBJECTIVE: To determine the levels of asthma knowledge in parents of asthmatic children and factors that may influence it. METHODS: This is a prospective study done between March 1998 and July 1998. Sixty-seven parents were interviewed using the 31 item asthma knowledge questionnaire that had been validated and translated. The children' asthma severity was classified. The questionnaire includes bio-data of children and parents, types of medication and dosages duration of asthma, exposure to cigarette smoke, acute asthma admissions, and parent's economic status. RESULTS: The mean score for asthma knowledge was 15.5. The total score was 31. Asthma knowledge was significantly higher in parents whose children were using steroids [p = 0.03, CI (-3.58, -0.02)]. It correlated significantly with steroid dosage (r = 0.29, p = 0.02), and was significantly higher in parents of higher economic status. Parent's asthma knowledge had no association with children's asthma status, age of the child or parents, exposure to cigarette smoke, frequency of admission or asthma duration. CONCLUSION: The low asthma knowledge level indicates the need to increase the effort in educating parents. The main indicator for higher knowledge was steroid usage and dosage. Higher asthma knowledge in the high-income group was probably related to levels of education. PMID- 12733174 TI - Do paediatric research presentations at local meetings get published? AB - Accessibility of research done locally to clinicians remains limited unless it is in the published form. The publication rate of research presentations at the Annual Malaysian Paediatric Association, Perinatal Society of Malaysia and Academy of Medicine Malaysia in 1997 and 1998 was determined. One hundred and five (95.5%) of 110 research presentations were carried out in Malaysia. Thirty seven (35.2%) presentations were published. University-affiliated institutions were more likely to publish their research presentations as compared to Ministry of Health hospitals (OR 3.1 95% CI 1.4-6.8, p < 0.01). There is a need to encourage publication of local research presentations. University-affiliated institutions performed better due to institution pressure for career advancement. PMID- 12733175 TI - Septooptic dysplasia. AB - The term septooptic dysplasia was coined in 1956 by de Morsier, who pointed out the association of optic nerve hypoplasia and absence of the septum pellucidum. Patients with this condition may present with clinical features of hypopituitarism, decreased visual acuity and neurodevelopmental disabilities that lead to this diagnosis. The case that is presented here is unusual in that this patient was initially diagnosed as having low tension glaucoma during a routine screening examination and was treated for glaucoma for over a year before he was discovered to have septooptic dysplasia, also known as de Morsier's syndrome. PMID- 12733176 TI - Optic neuritis and retinal vasculitis as primary manifestations of systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is a common multisystem disorder. However, retinal vasculitis as a primary manifestation of SLE is uncommon, accounting for only 4% of causes of retinal vasculitis. The postulated mechanism appeared to be vaso-occlusion of the retinal arterioles by thrombosis, with resultant ischaemia. Optic neuropathy in SLE is also rare, with a prevalence of 1%. This is a case report of a young lady who presented to us with retinal vasculitis as her initial presentation of SLE. Interestingly, the pathologic mechanism appeared to be inflammatory and not vaso-occlusive. PMID- 12733177 TI - Surgical removal of inverted papilloma: case report of endoscopic versus open excision. AB - We describe two cases of nasal inverted papilloma who had different management modalities. Although both were essentially of the same pathology, one underwent endoscopic excision while the other was treated with the more popular lateral rhinotomy and medial maxillectomy. We wish to highlight the different modalities of treatment available for inverted papilloma and a review of published results. PMID- 12733178 TI - Anterior shoulder dislocation with axillary artery and nerve injury. AB - We report a rare case of left axillary artery injury associated with anterior dislocation of the left shoulder in a 25 yrs old male as a result of a road traffic accident. The shoulder dislocation was reduced. A left upper limb angiogram showed an obstructed left axillary artery. The obstructed segment was surgically reconstructed with a Dacron graft. Six months post operation in follow up, he was found to have good left shoulder function and no neurovascular deficit. This is an injury that could have been easily missed without a simple clinical examination. PMID- 12733179 TI - Case of a non-pulsatile groin swelling. AB - We report a case of a non-pulsatile groin swelling in a 38 years old male drug addict without the typical clinical signs of an aneurysm. Ultrasound revealed a left femoral artery pseudo-aneurysm. He was surgically treated and the vessels were ligated without revascularisation. PMID- 12733181 TI - Hypertension update: lessons from the literature. PMID- 12733180 TI - Endoscopic repair of spontaneous cerebro-spinal fluid rhinorrhoea: a report of 3 cases. AB - Three cases of spontaneous Cerebrospinal Fluid (CSF) rhinorrhea were managed at the National University Hospital, Kuala Lumpur. Case 1 had bilateral leak secondary to empty sella syndrome and the rest two cases had unilateral leak. Four transnasal endoscopic approaches were performed on these three cases since March 1999. The role of intrathecal Sodium Fluorescein is highlighted in localising the CSF fistula. Case 3 required postoperative lumbar drain as an adjunct. No recurrent leak was noted on post operative follow up in Case 2 and 3 ranging from nine to thirty two months. A recurrent left leak at six months was noted in Case 1 which could likely be due to her sudden bout of cough attacks and patient refused further surgical intervention. PMID- 12733182 TI - Assessment and management of chronic cough. PMID- 12733184 TI - The future of medical education in Malaysia. PMID- 12733183 TI - Medical schools in Malaysia--meeting the future. PMID- 12733186 TI - Medical education: University of Malaya's experience. PMID- 12733185 TI - Medical education in Universiti Sains Malaysia. AB - Universiti Sains Malaysia established it's medical school in 1979, the third medical school in Malaysia after Universiti Malaya and Universiti Kebangsaan Malaysia. During the time of its establishment, the university was fortunate to witness a revolution in the world of medical education. PBL-based education was one of the most talked about approach in medical education. The University was fortunate to have experienced medical educators with sufficient foresight to start a medical school that has in its philosophy a community-based integrated curriculum utilizing problem-based learning, one of it's main modes of curricular implementation. Over the last 20 years, the medical curriculum has been revised and fine-tuned twice. The first major curriculum review was undertaken in 1995. One major outcome of this review was a firm commitment to continue with it's original philosophy in medical education at the same time introducing several key strategies to enhance the teaching of medical ethics, attitude formation and reaffirming the need for a lean, integrated curriculum which addresses core knowledge, attitude and skills. A more recent review in 2001 took several approaches including getting the input of students to enhance the original philosophy. PMID- 12733187 TI - Towards better quality medical education in Malaysia. PMID- 12733188 TI - The undergraduate ENT medical curriculum: past, present and future. AB - The strong international demand for admission into medical schools make medical education a "seller's market", and increasingly a global market. Teaching of Otorhinolaryngology-Head and Neck Surgery (ORL-HNS) has two primary goals. Firstly, a firm grasp of basic principles, recognition and treatment of common disorders, initial management of ORL-HNS emergencies and indications for specialist referral. Secondly, to provide sufficient exposure to the specialty to assist in career planning. Good communicative skills for optimal patient care are essential in the selection criteria of medical students. Proficiency in English is essential to obtain a disproportion share of opportunities in the new economy. The examination evaluation needs to be standardized between the various medical schools and the recommended lecturer-student ratio is maintained. The Joint National Evaluating Board has a very essential role to play in the maintenance of medical educational standards in Malaysia. PMID- 12733189 TI - Teaching and training in orthopaedics at undergraduate level in Malaysia: the consumers' viewpoint. PMID- 12733190 TI - Care of the older person in the medical curricula--the challenges. PMID- 12733191 TI - The growing importance of mental health: are medical curricula responding? AB - Mental health is becoming an important issue. Several local and international studies have proven that the incidence of mental illness is on the rise. Doctors have also been able to make more accurate diagnoses and treat mental disorders more reliably with the aid of recent research and newer drugs. As such it is necessary for the medical curricula to respond to this shift. Medical students must now be exposed to new psychiatric disorders and ways of managing them. The time spent in psychiatry and the mode of teaching must also be revised and modified to the current needs of patients. PMID- 12733192 TI - Is input on Complementary and Alternative Medicine (CAM) necessary in our medical curriculum? A perspective from a survey on medical practitioners. AB - A cross-sectional study was carried out through a postal survey of a random sample of registered medical practitioners in Malaysia to explore the pursuit and practice of CAM among them. A response rate of 42% was acquired. 27.1% of the medical practitioners are currently using CAM on themselves or their own families and 22.2% actually have referred patients to CAM practitioners. Analysis showed that only 14.9% of the medical practitioners who responded were exposed to CAM during their undergraduate days. Out of 28 respondents graduated from USM, 15 (53.6%) were exposed while out of the 80 graduates of UM, only 6 (7.5%) were exposed and out of 58 respondents graduates of UKM, only 5 (8.6%) were exposed to CAM during their undergraduate teaching. These differences are statistically different (p < 0.001). Analysis also showed that more (72.6%) medical practitioners are for having training in CAM during the medical undergraduate studies. Only 9.1% of the respondents have attended any training in CAM post graduation and 36.8% would like further training on CAM postgraduate and would pay for it. The findings illustrate the need for training in CAM in medical undergraduate education especially in this new age where alternative therapy is in demand by the consumers. PMID- 12733193 TI - Teaching medical students how to take a sexual history and discuss sexual health issues. AB - BACKGROUND: Taking a sexual history and discussing sexual health issues with patients form an important part of a medical consultation. These specific communication skills can be acquired through various teaching methods. OBJECTIVE: This paper describes the communication skill workshops conducted for undergraduate medical students on how to talk to patients about sex. METHODOLOGY: 198 medical students participated in a series of workshops conducted in the University of Malaya in 2001-2002. Pre- and post-workshop evaluations of the programme were carried out to find out the students' difficulties and to assess the usefulness of the workshop. The workshop consisted of a short lecture, role plays and discussion. RESULTS: Only 34% of the participants had received some informal training during their clinical years. The main barriers encountered were gender and age differences, language and choice of words, patients and doctors feeling shy, and cultural differences. The workshop was felt to be useful (mean score 4.38, maximum 5.0), most students felt comfortable during the workshop (mean score 4.10, maximum 5.0) and there was significant improvement in the "comfort level" when talking to patients about sex after attending the workshop (P < 0.001). CONCLUSION: Gender, language and cultural differences were the main barriers in taking a sexual history and discussing sexual health issues among the medical students. Communication workshop was felt to be a useful and comfortable method of learning these specific. PMID- 12733194 TI - Preparing for and managing change: some thoughts for curriculum planners and implementers. AB - This paper outlines issues related to curricular reforms, and strategies to be considered for planning and implementation so as to ensure that the change is institutionalised. In Malaysia, in general, some imminent curricular changes have been carried out to prepare graduates for future changes in the practice environment. Change of reform requires planning, with consideration of the directions in which the change is needed, and the possible educational approaches to be utilized. To ensure change can occur and be maintained there should be effort to induce a paradigm shift amongst teachers and administrators at all levels, there must be transparency and dissemination of information of the required change and why, there must be involvement of teachers in decision making to ensure better compliance. PMID- 12733196 TI - Ethics--the role of medical education. PMID- 12733195 TI - Utilization of information technology in medical education: a questionnaire survey of students in a Malaysian institution. AB - OBJECTIVE: The objective of this survey was to obtain a self-reported assessment of the use of Information and Communication Technology (ICT) by medical students at the International Medical University, Malaysia. MATERIALS AND METHODS: Students' perceived skills and extent of usage of ICT were evaluated using a questionnaire. Chi-square analysis were performed to ascertain the association between variables. Further statistical testing using Chi-square test for trend was done when one of the variables was ordered, and Spearman rank correlation when both variables were ordered. RESULTS: Overall, (98%) of students responded to the questionnaire. Twenty seven students (5.7%) did not use a computer either in the university or at home. Most students surveyed reported adequate skills at word processing (55%), e-mailing (78%) and surfing the internet (67%). CONCLUSION: The results suggests that in order to increase the level of computer literacy among medical students, positive steps would need to be taken, for example the formal inclusion of ICT instruction in the teaching of undergraduate medicine. This will enhance medical students' ability to acquire, appraise, and use information in order to solve clinical and other problems quickly and efficiently in the course of their studies, and more importantly when they graduate. PMID- 12733197 TI - Communication skills teaching in primary care medicine. AB - The teaching of clinical communication skills to undergraduate medical students in the Faculty of Medicine, University of Malaya is described. It is a continuous process throughout the five-year medical curriculum which is divided into Phases I, II and III. Students are introduced to communication skills early in Phase I through an interactive session as well as a workshop on general communication skills. In Phase II, small-group two-day workshops cover the basic principles of clinical communication skills using videotapes, group discussion and role-plays. Direct contact between students and patients in actual clinical setting begin in Phase IIIA. Communication skills teaching with feedback training is carried out by videotaping the consultations. In Phase IIIB the two-way mirror is utilized as well as having workshops on certain difficult areas such as 'breaking bad news' and 'taking a sexual history'. Formal assessment is done by evaluating the behavior, language and actual interview content. PMID- 12733198 TI - Critical appraisal--is there a need to train medical students how to read the literature? AB - Critical appraisal refers to the skill of reading a piece of research in a very objective and structured way. It allows for the reader to assess the quality and validity of the evidence put forward. With the emphasis on evidence-based practice in the medical profession, the ability to critically appraise the literature should be instilled into medical students. Currently, the push to encourage research shows great effort in the medical curriculum, through the incorporation of elective research programmes, by many medical institutions. But how ready are the students to even understand the research literature, let alone conduct a research? The current system throws these students into 'the deep end' of research conduct without equipping them with the tools necessary to do so. Very often this becomes a problem that snowballs through specialist training right up to the practice of medicine. The possibilities and means of introducing the skills of critical appraisal via the curriculum should be explored. In this age of self-directed and problem-based learning, a purely didactic teaching method of "how to read the literature" is surely outdated. The concept should be integrated into medical teaching, including within the implementation of the PBL system, in both the clinical and non-clinical settings, and by the introduction of the 'journal club' concept. Training of the trainers should also be considered. With the early training of critical appraisal, it can hopefully become an unconscious competence of medical graduates, who not only can produce quality research, but also able to identify quality information. PMID- 12733199 TI - Primary care medicine training in the undergraduate curricula--the UM experience. PMID- 12733200 TI - The Community Family Case Studies Programme at the University of Malaya. AB - The New Integrated Curriculum at the Faculty of Medicine, University of Malaya, Which comprises three major longitudinal strands, was first implemented in 1998 to provide better integration of clinical and basic medical sciences. One of these longitudinal strands, the Doctor, Patients, Health and Society (DPHS) module, emphasizes the importance of developing good communication skills as well as introducing students to behavioural sciences, public health medicine, statistics and epidemiology. Community Family Case Studies (CFCS), within this module, have been used as a means for students to focus on these aspects, as the students are introduced to patients, their families and their community in Year 1 of the medical course and are required to follow them up throughout their five year training period. PMID- 12733201 TI - Community Residency Programme (CRP)--a tool for research and rural health training for medical students. AB - Rural health training is an important element in the training of medical students in the University of Malaya. There is a need for the undergraduates to be familiar with the rural health infrastructure and to understand the social and economic aspects of the rural poor. The objective of the training is to make the students understand the problems faced by the poor in the rural areas so that when they practice in rural health areas, after graduation, they will understand the problems of the rural poor. They will have the knowledge of the diseases in the rural areas and also understand the community and the environmental factors that contribute to the disease. The training lasts' for 4 weeks, one week for lectures on health survey, two weeks for the field trip and one week of data analysis and presentation of their findings to an expert panel. During the field trip the students are divided into groups and they go to different parts of the country. Each group will do a field survey to find out the socio-demography, environmental, economic, nutritional and health problems in the village. In addition to the survey they also do a research project on any topic. The students also do social work, visit places of public health interest like the water treatment plant, sewage disposal, factory visits and others. Apart from technical skills in statistics and epidemiology, various other managerial skills like leadership, teamwork, communications and public relations are also learnt during the training. In conclusion this rural health training is an important aspect of the medical students training as it imparts several skills to them that are needed as a doctor. PMID- 12733202 TI - Introduction to management: Organization and Management of Health Services (OMHS) in Malaysia. PMID- 12733203 TI - Research training as pillars towards undergraduate medical education advancement in Malaysia. PMID- 12733204 TI - Teaching of evidence-based medicine to medical undergraduates. AB - Medical practice is changing, and the foundations of the paradigm shift lie in the development in research over the last four decades. Today, it is no longer adequate to treat a patient purely on clinical experience alone without a clear demonstration of evidence based on research, particularly the use of randomised controlled clinical trials. What is thought to be an effective mode of treatment currently may not necessarily hold true by the time medical students begin his/her medical practice. As a consequence, many medical schools worldwide are increasingly promoting evidence-based medicine (EBM) teaching in their medical curriculum along with problem-based learning (PBL). Teaching of EBM requires a paradigm shift in itself, as students must possess additional skills that are not traditionally part of medical training. These include the ability to acquire the skills in 'means of answering questions' than just 'knowing the answer to questions'. This paper aims to describe what EBM is and to highlight the formative experience of the teaching of EBM at the medical undergraduate level in the University of Malaya. Challenges and opportunities towards successful adoption of evidence-based practice are discussed. PMID- 12733205 TI - April Fool's Day has moved.... PMID- 12733206 TI - Delayed presentation of traumatic rupture of the diaphragm. AB - Ruptured diaphragm as a result of blunt trauma can present acutely or late in the disease, process. Late presentation is often a result of herniation of abdominal contents into the thorax. Patients may present with nonspecific symptoms, and may complain of chest pain, abdominal pain, dyspnea, tachypnea, or cough. Clinicians must have a high index of suspicion for prompt diagnosis. Diagnostic tools include chest radiograph, CT scan, and MRI. The treatment for rupture of the diaphragm is surgical. The authors report a case of traumatic rupture of the diaphragm presenting 20 years after an automobile accident with blunt trauma to the abdomen. PMID- 12733207 TI - Celebrating the structure of DNA: 50 years and beyond. PMID- 12733208 TI - Acute coronary syndrome associated with coronary aneurysm: a case report. AB - An elderly female patient presented to our Emergency Department with an acute coronary syndrome. After stabilization, cardiac catheterization disclosed a solitary aneurysm of her diagonal coronary artery. Such aneurysms are rare and are an unusual etiology for acute coronary syndrome. PMID- 12733209 TI - Prevention of lawnmower injuries in children. AB - While we can legislate all we want and devise countless safety devices, the fact remains that the primary responsibility must be with parents and families. Perhaps the only way this can be accomplished is with a vigorous public awareness and education campaign. If a parent or grandparent could see the devastation caused by these injuries to other children, perhaps they would think twice before allowing a child to operate a power mower or to be a passenger on a riding mower. Young children should never be allowed to play in the vicinity of a person who is operating a mower. Physicians should be involved in this education effort. These injuries are preventable. PMID- 12733210 TI - Baseball safety for children. PMID- 12733211 TI - A year in providence. PMID- 12733212 TI - Partners in time. PMID- 12733213 TI - The HSJ interview: Pat Troop. Soldiering on. Interview by Graham Clews. PMID- 12733214 TI - Older people's services. A picture of health. AB - A scheme offering joint health and social care assessments to people aged over 75 in inner London identified more than a quarter in need of services. The older people were accessed via general practice, but this proved an inefficient way of identifying the target population. In some practices, almost half of those contacted were no longer at the same address. The assessments, conducted by a community nurse and social welfare officer, often involved two visits. The mobility of the elderly population needs to be taken into account when planning joint assessments. Organisations involved had different aspirations for the scheme and management proved problematic. PMID- 12733215 TI - Disability access. Open season. AB - A disability access audit carried out at a trust operating over 50 sites revealed that a 2.3m Pounds programme of work was needed. The audit took four months, with the team spending a day at each of the premises. The audit has been followed by a staff training programme in disability awareness. The trust's information systems now show if a patient did not attend an appointment because of difficulties with physical access. All letters to patients are produced in a minimum 12-point type. PMID- 12733216 TI - User involvement. Pick and choose. AB - Involving service users in the selection of a manager for learning disabilities services proved successful when implemented by a primary care trust with the use of facilitators and special training. The PCT chief executive made it clear that no-one would be appointed if the user and carer panel considered them inappropriate. The users were involved in drawing up five questions for each of the candidates. On the basis of their answers, each candidate was scored by the service users. The users, carers and managers were unanimous in their choice of candidate. PMID- 12733217 TI - Are you losing staff to military call-ups? ED managers share their key strategies. AB - Many EDs are losing staff due to military call-ups, and the problem is expected to worsen in the coming months. Use flexible staffing including short shifts. Develop a pool of volunteer ED physicians willing to work per diem. Plan for solutions in advance with administrators. PMID- 12733218 TI - Address risks of staff who get smallpox vaccine. AB - You'll need a policy to reduce risks of inadvertent transmission of the smallpox virus to address staff participation in the military's vaccination program. Vaccinated staff must wash hands, especially after touching the vaccination site or bandages. Implement Centers for Disease Control and Prevention guidelines to avoid transmission to staff, patients, and visitors. Have newly vaccinated staff avoid contact with immunocompromised patients if possible. PMID- 12733219 TI - 5 ways to comply with HIPAA oral privacy regs. AB - ED managers should use cost-effective and simple strategies to comply with Health Insurance Portability and Accountability Act requirements for oral privacy. Consider using cubicles or screens to block sound, and perform triage in a room next to the waiting area. Limit the number of visitors per patient, and require badges. Install white noise machines, sound-absorbent curtains, and ceiling tiles with a higher noise-reduction rating. PMID- 12733220 TI - Use protocol to send inpatient holds upstairs. AB - A "full capacity" protocol requires that admitted patients being held in the ED are moved upstairs when the ED is at full capacity. Length of stay for admitted patients is reduced. Morale of ED nurses is improved. Inpatients receive better care when moved upstairs. PMID- 12733221 TI - New strategies assist billing under APCs. PMID- 12733222 TI - Three legal frameworks for regulating genetic technology. PMID- 12733223 TI - Drugs and vaccines for the common defense: refining FDA regulation to promote the availability of products to counter biological attacks. PMID- 12733224 TI - Botanical drugs: a future for herbal medicines. PMID- 12733225 TI - Bad habits: the qui tam provisions of the False Claims Act are unconstitutional under Article II. PMID- 12733226 TI - Genetic screening: an employer's tool to differentiate or to discriminate? PMID- 12733227 TI - An argument for considering parental smoking in child abuse and neglect proceedings. PMID- 12733228 TI - The legacy of too little, too late: the inconsistent treatment of postpartum psychosis as a defense to infanticide. PMID- 12733230 TI - Subrogation or subterfuge: the myth of ERISA health benefit plans. PMID- 12733229 TI - Let him die with dignity or hope for a cure: the consequences of modern medicine. PMID- 12733231 TI - Aging and in search of healthcare: access to employer-provided retiree health benefits would be damaged for the boomer generation under an Erie County interpretation. PMID- 12733232 TI - Daimonic elements in early trauma. AB - This paper explores some of the 'daimonic' elements of unconscious mentation that emerge both in dreams and in the transference/countertransference field with early-trauma patients and illustrates these with an extended clinical example. An archaic and typical (archetypal) 'trauma complex' is articulated (with diagram) as a bi-polar structure consisting of divine child protected and/or persecuted by an inner 'guardian angel'. Sources of this structure and its mythological inner objects are traced to trauma at the stage of what Winnicott calls 'unintegration' and to flooding by disintegration anxiety at a time before nascent ego-structure has formed. In an extended case example, the author shows how the patient's traumatized innocence and desire for a new start, thwarted by self-attacking defences, pulls him into playing the inflated role of her guardian angel, leading to re-traumatization in the transference. Working through is seen as the necessary disillusionment and humanization of these daimonic structures as they are projected, suffered, and transmuted by the analytic partners in the stormy process of psychotherapy. PMID- 12733233 TI - Developmental aspects of trauma and traumatic aspects of development. AB - In this paper I try to show that inherent in healthy development are emotional experiences that are the stuff of trauma. Failure to reintegrate these experiences means that they can serve as a resonating board for difficulties in later life, adding to their traumatic impact. Focusing on global changes that occur at the end of the first year, I exemplify these developments with an infant observation and show how a six-year-old boy's failure to integrate them contributed to his experiencing a normal life event as a trauma. I then offer clinical material from the analysis of a man to demonstrate how later life events resonated with early experience associated with this period. All are linked to a complex pattern of object relations I have come across clinically, whereby feelings of grief associated with an idealized object are split off from feelings of grievance against an object experienced as persecutory because of its perceived superior status. PMID- 12733234 TI - Trauma and defences: their roots in relationship. An overview. AB - In this paper the differing psychodynamic models of defences are outlined and compared with an attachment theory perspective in which affect regulation plays a central role. Behavioural and intrapsychic distance regulation (defensive exclusion) are seen as the main strategies for affect regulation and are the manifestations of the habitual pattern of emotional regulation in the relationship between the child and the primary caregiver. A new perspective on unconscious fantasy is offered, in which fantasies are seen to be actively created as defensive narratives to protect the development of healthy narcissism and to become integrated into a person's internal working models. Archetypal defences are explored from a developmental perspective and some neurobiological issues relevant to defences are highlighted. PMID- 12733235 TI - Undoing trauma: contemporary neuroscience. A Jungian clinical perspective. AB - This paper uses insights from contemporary neuroscience and attachment theory to explore the profound dissociative defences associated with trauma. I discuss the effects of trauma on the emotional, intellectual and imaginative life of the individual and on the development of the self. Based on work with three patients with very different experiences of trauma, the paper offers clinical illustration of 'right brain to right brain' Jungian analysis. I argue that through repeated transference and countertransference experiences dissociative defences may be undone and change brought about. PMID- 12733236 TI - Incest in Jung's work: the origins of the epistomophilic instinct. AB - In this paper the differences between Jung's and Freud's writings on incest are explored. Jung's view is that the purpose of the child's sexual interest, as expressed also in his incestuous longings, is not purely the satisfaction of the biological instinct but is more importantly seen to be the development of thinking. The importance of the incest taboo for analytic work and the dangers of enactment of the erotic transference-countertransference dynamics are highlighted. PMID- 12733237 TI - A trajectory of work and success. PMID- 12733239 TI - [Risk of HIV infection: how women living in a slum perceive themselves in the chain of the virus transmission]. AB - The number of notified cases of women with AIDS is increasing in Brazil. Based on this, the authors developed this qualitative study theoretically based on Cultural Anthropology. The ethnographic methodology was used to collect and to analyze data. The investigation had the purpose to identify the perception about the risk for HIV infection of the women living in a slum located in the city of Sao Paulo, Brazil, through interviews and participant observation of the context. Results showed that the women have some knowledge about AIDS, about the transmission of HIV and its preventive measures through sexual intercourse. The cultural data also showed that the women do not feel susceptible to the disease, although most of them are dependent on their partners' income to maintain themselves healthy as well as their families. This way of thinking and acting may be contributing to increase the cases of women with AIDS in Brazil. PMID- 12733238 TI - [Depression: viewpoints and knowledge of the nurses in th basic health network]. AB - This study aimed at identifying the viewpoints and the knowledge on depression of nurses working in the basic health network. The 73 participants who answered two self-applicable questionnaires were distributed in 28 health units. The general results concerning their viewpoints and knowledge are, in most answers, in accordance with what was expected and showed average knowledge related to depression. However, the individual analysis of such answers indicate that these professionals are not in direct contact with and do not know how to identify depressed patients. They do not observe these indicators in the patients that they assist or do not understand that it is their task to do so. Reflections appeared concerning the possible relationship between mental health actions and their professional education. In this way, undergraduate and continuing education become more important in the development of nursing actions for mental health. PMID- 12733240 TI - [Child care according to day care center workers]. AB - This investigation aimed at understanding and analyzing the representations of the workers from daycare centers regarding the care they provide to children. Data were collected through individual interviews and an educational workshop. These interviews were carried out in daycare centers, that are associated to a public university in the municipality of Sao Paulo, Brazil. The topic presented here, Childcare in daycare centers, is the first result of this study. The care was defined as actions taken to attend to the child's physical and emotional necessities, a preliminary step to pedagogical activities and as a learning moment for children. The workers understand the care as a phenomenon that do not need specific training or preparation as it is secondary if compared to education. This common sense understanding can be a result of the lack of reflection about a theoretical and conceptual reference that must base the performance of these workers. PMID- 12733241 TI - [Burn prevention: perception of the patients and their relative ]. AB - This study aimed at investigating the ways to prevent burns identified by burned patients and their relatives. Data were collected at the Burns Unit of the Ribeirao Preto Medical School Clinical Hospital, University of Sao Paulo--Brazil by means of interviews with burned patients and their relatives. Four referees categorized data. In order to verify the existence of agreement among the referees with regard to categorization, the non-parametric correlation measure was used--Contingency Coefficient. Fifty-seven people were interviewed: 24 relatives and 33 burned patients. Of the 33 interviewed patients, 18 had suffered accidents at home. Of the 57 interviewees, 11 stated that they could not have avoided the accident causing the burn. Twenty patients and 12 relatives identified risk situations at home or at work and 13 patients and 12 relatives did not identify any types of risk situations. The relatives and patients referred to the following preventive measures for burning accidents: to be attentive to the activities that they perform and to be more careful while handling flammable products. PMID- 12733242 TI - [Gastric cancer: risk factors in patients treated in tertiary care services of a municipality in the interior of the state of Sao Paulo]. AB - This study was developed with gastric cancer patients submitted to surgery with the purpose to identify their exposure to risk factors for the disease. Twenty four clients who had agreed to participate by signing an informed consent term and responding a semi-structured instrument elaborated according to the Health Field perspective were the subjects in the study. The data analysis allowed to identify that 50.1% referred to inadequate diet, smoking, alcoholism and psychological problems as the responsible factors. With regard to the consumption of food considered to be harmful, there was a tendency to larger consumption of canned food, sausages, smoked and barbecued meat and fried food in addition to diets with high sodium content. 66.7% smoke or used to smoke and 70.8% indicated the consumption of alcoholic drinks, preferably beer or distilled types. These data show the need to implement educational programs with the purpose to inform about risk factors aiming at health promotion. PMID- 12733243 TI - [The participation of women with breast cancer in choosing their treatment: a right as yet to be gained]. AB - The purpose of this study was to identify how women with breast cancer perceive themselves as subjects in the process of making decisions on their own treatment. Two objectives were pursued: (a) to identify social and political determinants that, by affecting the socialization process of these women, might have influenced them in adopting a style of participation, and (b) to understand the meaning of such participation, as it was perceived by these women at the time they decided on their treatment options. The theoretical-methodological support adopted was that of Symbolic interactionism. The population sample included nine women with breast cancer. Semi-structured interviews allowed data to be gathered, and led to collecting further field notes and data from medical records. The hermeneutic dialectic method was employed as a compass for data interpretation, which made it possible to identify two broad theme units: 'construction of the female identity' and 'style of participation when choosing one's own treatment'. By means of these units it was possible to grasp what it meant for these women to deal with the issue of limits and, therefore, of ethics. In their view, they did not participate in the decision-making process, being regarded as unqualified to decide on the fate of their own bodies and lives. They were thus seen as obedient subjects in relation to medical decisions that are based on the principle of beneficence, where health care delivery is dependent on hierarchical social relationships and power structures are present between classes, genders, and levels of knowledge. PMID- 12733245 TI - [Daily routine of families with cystic fibrosis children: subsides for pediatric nursing]. AB - Cystic Fibrosis is a genetic disorder characterized by the increase in the production of mucus whose accumulation on some organs causes: chronic obstructive pulmonary disease, pancreatic insufficiency and a high level of electrolytes in sweat. This study aims at describing the routine of families with children bearing cystic fibrosis as to the aspects concerning the effects of that chronic disease on the family dynamics. To that end, a qualitative study was conducted. Empirical data were collected by means of interviews with the families of patients aged less than 18 years old undergoing treatment at Hospital das Clinicas da Faculdade de Medicina de Ribeirao Preto da Universidade de Sao Paulo. The following themes emerged from the collected data: family involvement, acceptance/understanding of the disease and coping with challenge. A dependent life was identified which was related to the fact that the disease caused physical waste and emotional disturbance for both the child and the family. IMPLICATIONS FOR NURSING: constitution of care models focusing on the family by considering the environment, life style and health as its basic fundamentals. PMID- 12733244 TI - [Functional evaluation of elderly patients with lower limb amputation followed at a university hospital]. AB - We studied 40 elderly with lower limb amputation (above the malleolus), who attended a hospital outpatient clinic (Orthoses and Prostheses Unity, University Hospital, State University of Campinas), from June 1994 to June 1999. Our objectives were to evaluate the independence level according to the Barthel's Index and to identify relationships between the mentioned Index and the type and level of amputation, as well as the prosthetic use. The subjects had a high degree of independence, according to the Barthel's Index. We found statistical significance only for the relationship between the Barthel's Index and the type of amputation, suggesting that the elderly with unilateral amputation were more independent than those who had undergone bilateral amputation. PMID- 12733246 TI - [Beliefs of patients with coronary disease, according to Rokeach theoretical framework, concerning the smoking habit]. AB - This is a descriptive observational study that aimed at identifying beliefs according to centrality/periphery by verifying how they interfere in the behavior of patients with coronary diseases regarding their adherence or non-adherence to smoking. The belief-related frameworks mentioned in the literature were used and findings, pointed out from the analysis of the interviews with 56 people with coronary diseases who were inpatients in a hospital in the interior of Sao Paulo State, showed a prevalence of primitive beliefs with zero consensus and a tendency to centrality that enabled the identification of hiding factors to the adherence to therapeutic recommendations by health professionals as well as inferences concerning the reluctance to behavioral changes presented by the population. PMID- 12733247 TI - [Nurses attitude toward the manual recording system of medical records]. AB - The main goal of this work was to identify factors related to nurses' understanding about the manual recording system, through a descriptive exploratory approach, developed with two hundred nursing professionals working at different hospital institutions located in the city of Joao Pessoa/Paraiba, Brazil. Data were collected with special forms using the Likert's behavioral scale. Results identified five factors such as: a) lack of knowledge about information systems; b) dissatisfaction with the manual recording system; c) needs of an informatized recording system to be used in nursing care; d) desire to utilize a classification system with all steps of the nursing process; and e) difficulties with nursing diagnosis related to manual recording. Findings demonstrated the dissatisfaction of nursing professionals about the manual recording system, requiring deep changes in the established recording model. PMID- 12733248 TI - [Drug therapy orphans: the administration of intravenous drugs in hospitalized children]. AB - Descriptive study, developed at a general university hospital that aimed at verifying the number and types of i.v. drugs administered to children, the adequacy of their pharmacological presentation for pediatric use and the estimated costs of some drugs administration. In a period of 30 days, 8,245 drug doses were administered, with an average of 274.83 doses a day, and a yearly estimation of 98.940. The most used drugs were methylprednisolone, vancomycin, furosemide, ranitidine, penicillin, amikacin, midazolam, fentanyl, ceftriaxone, cephalothin, oxacillin, ampicillin and metronidazole. None of the 41 different drugs had a pediatric presentation, what caused, in some cases, more manipulation during the preparation, increasing the contamination risks and the loss of stability. Authors observed that the lack of pediatric presentation generated an increase in care costs; as an example, considering the prescription of a child in the period after surgery, with an estimated time of hospitalization of 5 days, the daily therapy costs were of U$6.71, and U$39.52 of drugs were thrown away as they exceeded the children therapeutic needs. PMID- 12733249 TI - [Tuberculosis in the elderly: concept analysis]. AB - The goal of this study is to define the concept of tuberculosis in the elderly expressed in the literature covering the 80's and 90's. We have used concept analysis with emphasis on "essential attributes", "preceding events, and "consequential events", which build up the core sense of the concept to be defined. The elderly tuberculosis is expressed as the recrudescence of infection inactivated for a long time. The elderly is predispose to endogenous and exogenous reinfection. Among the most important "preceding" events are the institutional housing as a contact disease source. The "consequential" events are related to the need of a differentiate treatment and diagnostic in elderly due clinical conditions. We arrive at the conclusion that new studies about the concept analysis need to be developed in order to contribute to the knowledge about the diagnosis, treatment and care to elderly patients. PMID- 12733250 TI - [Diabetes mellitus educational software for health professionals: stages of design and development]. AB - This research aimed at creating a software about Diabetes mellitus in order to stimulate the continuing education of health professionals as well as educative practices. The system was developed using Delphi application, through a data bank utilizing key-words and the Pascal language. The software includes key-words searching tools, improving the process of accessing the data bank. PMID- 12733251 TI - [Hospital infection in the context of health policies instituted by the state of Santa Catarina]. AB - The Ministry of Health is guiding its health actions aiming at controlling and/or improving the care. Therefore, in 1994, a national study and research was developed in order to understand and evaluate the quality of the actions related to the control of hospital infection in the country. The results showed a medium incidence of hospital infection (15.5%) and a number of hospitals that correctly perform risk procedures (42.5%). The purpose of this study was to report the work of the Commissions for Hospital Infection Control in the context of the health policies established in the state of Santa Catarina, Brazil. The Program for Hospital Infection Control was created in 1992 and had a high receptivity, implementing commissions in 90 hospitals what represented a significant increase (from 7% in 1992 to 43% in 1993). There are in the state 177 Commissions for Hospital Infection Control, totaling 77%. Nowadays, 16 hospitals are accredited in infection control and 30 are being prepared for the accreditation evaluation. PMID- 12733252 TI - [Evidence based nursing: principles and applicability]. AB - The work considers the evidence based nursing as a new operational model that integrates individual clinical competencies, through clinical finding generated by scientific research and analyzes the inclusion of this practice to the actual nursing professional model. PMID- 12733253 TI - [Local complications in the skin related to the administration of insulin]. AB - Insulin administration is one of the important aspects in diabetes education. This study focuses on skin site complications related to insulin application. Fifty-one people with diabetes mellitus were interviewed at a private institution in Sao Paulo State. Results showed that hematomas were the most frequent complication and that they mainly resulted from the use of incorrect application techniques. It was concluded that health professionals must design effective strategies in order to provide safety to subjects and prevent skin complications. PMID- 12733254 TI - [Epidemiology of respiratory infections at institutional settings (screening and management)]. PMID- 12733255 TI - [Epidemic viral respiratory infections at institutional settings (flu and RSV). Epidemiology and clinical course of epidemic viral respiratory infections at institutional settings]. PMID- 12733256 TI - [Management and prevention of epidemic flu at institutional settings]. PMID- 12733257 TI - [Epidemic viral respiratory infections at institutional settings (flu and SRV): conclusions, synthesis, and perspectives]. PMID- 12733258 TI - [Epidemic bacterial respiratory infections at institutional settings (pneumococcus, legionella). Epidemiology and clinical course of epidemic bacterial respiratory infections at institutional settings]. PMID- 12733259 TI - [Management and prevention of epidemic bacterial respiratory infections at institutional settings]. PMID- 12733260 TI - [Epidemic bacterial respiratory infections at institutional settings: conclusions, synthesis, and perspectives]. PMID- 12733261 TI - [Epidemic tuberculosis infection at institutional settings. Epidemiology and clinical course of epidemic tuberculosis at institutional settings]. PMID- 12733262 TI - [Management and prevention of epidemic tuberculosis at institutional settings]. PMID- 12733263 TI - [Epidemic tuberculosis infection at institutional settings: conclusions, synthesis, and perspectives]. PMID- 12733264 TI - The comparative analysis of the myenteric plexus in pigeon and hen. AB - Using the thiocholine method and histological techniques the myenteric plexuses of pigeon and hen were studied. Investigations revealed the presence of a nervous network in the wall of the small intestine of both animals. It consists of many nerve fibres crossing each other and creating meshes in a variety of shapes. The density of the network was different according to the species and to the parts of intestine. The myenteric plexus from the pigeon's duodenum is thicker (3.7-fold) than in the remaining part of the small intestine; in the hen this is approximately 2.2-fold thicker. The meshes of the duodenum in both species are smaller than in the jejunum and ileum. The results of histological investigations showed different localization of myenteric plexuses; in pigeon in the space between the circular and longitudinal muscle layers, and in hen within the circular muscle. PMID- 12733265 TI - Arterial supply of the penis in the New Zealand rabbit (Oryctolagus cuniculus L.). AB - In the present study, the distributional pattern of the penile artery and the vessels joining the blood supply of the penis were investigated in the New Zealand rabbit. Eight adult rabbits were used in the study. In order to exhibit the vascular network by dissection, latex was injected via the abdominal aorta. The main vessel which supplies blood to the penis, the penile artery, is a branch of the internal pudendal artery. It divides into two branches which form the deep and dorsal penile arteries at the level of the ischiadic arch. The deep penile artery penetrates the tunica albuginea, and forms the arterial network of corpus cavernosum penis. On the other hand, the dorsal penile artery gives off three small branches for the subischiocavernosus muscle and at the level of the attachment of this muscle sends two small branches for the preputium. The course of both arteries follows the dorsolateral surface of the penis to the glans and ends in an anastomosis. Hence, a caudal branch of the prostatic artery which originates from the umbilical artery joins the blood supply of the penis in the rabbit. After vascularizing the prostate complex, it ends by entering the corpus spongiosus penis at the dorsolateral surface at the level of the ischiadic arch. PMID- 12733266 TI - Large interarcuate spaces in the cervical vertebral column of the tyrolean mountain sheep. AB - Large interarcual spaces have been described between the arcus vertebrae C5/C6 and C6/C7 in the cervical vertebral column of Nubian goats. This aperture enables direct access to spinal cord and rootlets without the need to perform a hemilaminectomy. The present study was performed in order to determine whether these large interarcual spaces can also be found in the vertebral column of the Tyrolean mountain sheep, as this small ruminant, which is anatomically very similar to the Nubian goat, is frequently used for experimental purposes at the Surgical University Clinic in Austria. The carcasses of 10 sheep (six females, four males; range of age: 2.5-6 years, range of weight: 52-89 kg) were dissected and the vertebral column was exposed. All 10 sheep showed elliptic openings between the fourth cervical and the first thoracal vertebrae. Three sheep had additional openings between the first and the second thoracal vertebrae. All openings were covered solitarily by the ligamentum flavum and under this ligamentum lay the spinal cord without any further osseous or ligamentous protection. These findings are not mentioned in the common textbooks of veterinary anatomy and deserve attention, as they can be a step forward towards non-traumatic experimental surgery on the spinal cord. PMID- 12733267 TI - Age-related changes in rat optic nerve: morphological studies. AB - Age-related changes of the optic nerve were studied in 3-month-old (young), 12 month-old (adult) and 24-month-old (aged) male Sprague-Dawley rats. Cross sections of the intracranial portion of the optic nerves of animals of different age groups were stained with haematoxylin-eosin and examined under a light microscope at low and high magnification. Other sections were stained with crystal violet for demonstration of glial cells. A third group of sections were stained immunohistochemically to detect glial fibrillary acidic protein (GFAP) which is a marker for localizing and characterizing astrocytes. All morphological results were subjected to the quantitative analysis of images and to statistical analysis to identify significant morphometrical data. Tissue protein concentrations were determined on homogenized fragments of optic nerve. Our results demonstrate the following age-related changes: (1) increase of the optic nerve sheaths (meningeal membranes); (2) increased number of astrocytes; (3) increase of areal density of GFAP immunoreactivity; (4) increased diameter and area of the optic nerve; (5) decreased number of nerve fibres; (6) decreased-size of nerve fibres and (7) decrease of the nerve fibres/meningeal membrane ratio from 3:1 to 1:1. Moreover, the protein amount does not change with age. The rat optic nerve, therefore, appears sensitive to ageing processes and can be considered as a useful model for the studies on neuronal ageing. PMID- 12733268 TI - Alcohol effects on the principal and clear cells of the caput epididymis of albino rats. AB - Ultrastructural observations of principal cells of the epithelium lining of the proximal caput epididymis in experimental alcoholic albino rats at 180 days of treatment showed pyknotic nuclei, ill-defined cellular organelles and clusters of electrondense bodies, perhaps lysosomes. It was also verified for a progressive accumulation of lipid droplets initially in the basal and perinuclear cytoplasm and finally in the apical cytoplasm of principal cells at 60, 120 and 180 days of experimentation, respectively. The clear cells of alcoholic rats at 180 days showed the cytoplasm totally filled with lipid droplets. These findings were taken comparatively with the morphological features of the same epididymal cells in control (normal) rats. PMID- 12733269 TI - An immunohistochemical study of the gastrointestinal endocrine cells in the C57BL/6 mice. AB - The regional distributions and relative frequencies of some gastrointestinal endocrine cells in the eight portions (fundus, pylorus, duodenum, jejunum, ileum, cecum, colon and rectum) of the gastrointestinal tract of C57BL/6 mouse was studied with immunohistochemical method using seven types of specific anti-sera against chromogranin A (CGA), serotonin, somatostatin, human pancreatic polypeptide (HPP), glucagon, gastrin and cholecystokinin (CCK)-8. In this study, all these seven types of immunoreactive (IR) cells were identified. Most of these IR cells in the intestinal portion were generally spherical or spindle in shape (open-type cell) while cells showing round in shape (closed-type cell) were found in the intestinal gland and stomach regions occasionally. Their relative frequencies were varied according to each portion of gastrointestinal tract. CGA IR cells were demonstrated throughout the whole gastrointestinal tract and they showed most predominant in the pylorus and duodenum. Serotonin-IR cells were detected throughout whole gastrointestinal tract and they showed highest frequency in the stomach and colon. Somatostatin-IR cells were demonstrated throughout whole gastrointestinal tract except for large intestine and showed highest frequency in the fundus. HPP-IR cells were found in the fundus with rare frequency. Peculiarly, glucagon-IR cells were restricted to the fundus, ileum and colon with a few frequencies. Gastrin-IR cells were restricted to the pylorus with numerous frequency and CCK-8-IR cells were observed in the pylorus, duodenum and jejunum with numerous and/or a few frequencies, respectively. In conclusion, some peculiar distributional patterns of gastrointestinal endocrine cells were found in C57BL/6 mouse. PMID- 12733270 TI - Localization of Neuropeptide Y-like immunoreactive structures in the brain of the pejerrey, Odontesthes bonariensis (Teleostei, Atheriniformes). AB - The aim of this study was to determine the distribution of Neuropeptide-Y (NPY) immunoreactive neurons and fibres in the brain and pituitary of Odontesthes bonariensis by immunohistochemical methods. A wide distribution of immunoreactive NPY (ir-NPY) cells and fibres in the forebrain and midbrain was observed. A prominent ir-NPY nucleus was found in the ventral telencephalon and other ir-NPY cells groups were recognized at the dorso-medial telencephalon. The diencephalon showed ir-NPY cells in the Nucleus entopeduncularis, the Nucleus preopticus periventricularis and in the Nucleus lateralis tuberis. Ir-NPY fibres were conspicuous in the preoptic region and the hypothalamus. There were also numerous ir-NPY fibres at the epithalamic level running ventrally to the hypothalamus and the pituitary stalk. At the rhomboencephalic level, the ir-NPY neurons were observed in the Locus coeruleus. Double-labelled immunostaining showed a close association between ir-NPY fibres that reach the adenohypophysis and growth hormone (GH)- and gonadotropin (GtH)-expressing cells. Although our results exhibit some relevant differences when compared to other fish groups, they support the existence of a conserved NPY system in teleosts. PMID- 12733271 TI - Post-natal maturation of acinar cells of the guinea pig pancreas: an ultrastructural morphometric study. AB - The morphological maturation of the acinar cells of the guinea pig pancreas during post-natal development was characterized morphometrically by determining the intracytoplasmic accumulation of rough endoplasmic reticulum (RER) and zymogen granules. The following results were obtained for the period analysed, i.e., from 2 to 70 days of post-natal life: (a) the acinar cell volume increased by 210% (P < 0.01); (b) the mostly cisternal RER occupied more than 30% of the cytoplasm at any age studied and their total volume and surface in the cell were increased by 300 and 534% (P < 0.01), respectively; (c) maturation in the morphological pattern of the RER was observed; (d) the mean number of zymogen granules per cell increased from 261 at 2 days to 422 at 70 days (P < 0.01), while their mean diameter increased from 0.52 to 0.94 micron (P < 0.01) during the same period; (e) these increases in granule number and size were responsible for a 500% (P < 0.01) increase in total volume from 2 to 70 days and for a 304% increase (P < 0.01) in total surface from 2 to 35 days; (f) the RER and the zymogen granules together occupied 44, 54, 55 and 57% of the cytoplasm at 2, 14, 35 and 70 days of age, respectively. We conclude that although the pancreatic acinar cells of the guinea pig are morphologically well differentiated at 2 days of age, with the cytoplasm already showing a large amount of RER and zymogen granules, they are still immature. Morphological maturation of the acinar cell occurs during the first months of post-natal life and is characterized by a substantial gain in cell volume and intracytoplasmic accumulation of RER and zymogen granules, which significantly increase of both their absolute volume and total surface, with a higher growth rate being observed during the period from 2 to 14 days of post-natal life. PMID- 12733272 TI - Morphological, morphometric and histochemical characterization of the gastric mucosa of the camel (Camelus dromedarius). AB - Mucosal morphology, morphometry and mucin histochemistry of the stomach were studied in the one-humped camel. The lining of the stomach was divided into eight grossly identifiable regions. The first region was non-glandular, occupied the body of the first compartment of the stomach and constituted 53.2% of the gastric mucosa. The other seven regions were lined by a glandular mucosa. Histological, histochemical and morphometric investigations have shown that glandular mucosa comprises pseudo-cardiac, cardiac, fundic and pyloric regions. The pseudo-cardiac region was characterized by widely separated short tubular serous glands. It constituted 36.2% of the gastric mucosa; it extended over the entire lining of the second compartment and parts of the first and third compartments. The cardiac region was confined to the initial zone of the third compartment amidst the psuedo-cardiac region but contiguous with the distal end of the gastric groove. It constituted 3.4% of the gastric mucosa and was characterized by neutral and acid mucin positive glands. The fundic and pyloric regions occupied the distal distended part of the third compartment. The fundic region constituted 4.3% of the gastric mucosa. It was packed with typical fundic glands characterized by chief cells, parietal cells and acid mucin positive neck cells. The pyloric region constituted 2.9% of the gastric mucosa. Its glands were positive to acid mucins except for their bases that were positive to neutral mucins. Differences in volume densities of the mucosal components and reactivity of the surface epithelium and gastric pits to mucin stains were noted in the different regions of the stomach. PMID- 12733273 TI - Apoptosis and proliferation during seasonal testis regression in the brown hare (Lepus europaeus L.). AB - Testes samples of 52 brown hares (Lepus europaeus L.), sacrificed between July and January, were subjected to immuno histochemical analysis. The terminal deoxynucleotidyl transferase-mediated d'UTP nick end labelling (TUNEL) method was applied to detect apoptosis; and antibodies to proliferating cell nuclear antigen (PCNA) were used to evaluate cell proliferation in the testes. In the seminiferous epithelium, the apoptotic processes were evident from August to early November with maximal values in September. Cell death in germ cells occurs predominantly during the prophase of the first meiotic division. In July, and from mid-November onwards, only the occasional TUNEL-positive cells can be seen. The proliferation of germ cells continues during the testis regression phase. The average number of PCNA-positive cells decreases slightly from September onwards and rises again in mid-November. PMID- 12733274 TI - Atrophic and regenerative changes in rabbit mimic muscles after lidocaine and bupivacaine application. AB - Destruction and denervation atrophy in skeletal muscles caused by the injection of local anaesthetics was investigated by injecting lidocaine or bupivacaine around the rabbit facial nerve to produce facial paralysis. Animals were then sacrificed at 2, 4, 6, and 8 weeks post-injection, and changes in mimic muscle tissue were assessed at each stage by light microscopy and electron microscopy. Atrophic changes were observed at 2-6 weeks after injection, and regeneration started at 6-8 weeks. Compared to bupivacaine, lidocaine caused more dramatic atrophic changes and was associated with slower muscle regeneration. PMID- 12733275 TI - Diprosopiasis in a lamb. A case report. AB - Conjoined twinning has been reported in most of the domestic animal species. Among them, sheep have the highest incidence of craniofacial defects. A live male crossbreed dystocic two-headed lamb was delivered from a 2-year-old Pinzerita sheep after first mating. After 40 h of life, the lamb spontaneously died. The most important gross findings involved the head, whereas the examination of different organ and tissue sections did not reveal remarkable histomorphological changes. The lamb was classified as a conjoined twinning and, on the basis of the facial duplication, as a diprosopus tetraophtalmus. PMID- 12733276 TI - US emergency nurses support colleague in child abuse case. PMID- 12733277 TI - New ways of cutting waiting time for primary care. PMID- 12733278 TI - The jungle nurse. PMID- 12733279 TI - Ambulance nursing assessment. Part two. PMID- 12733280 TI - Participation in warfare: a literature review. PMID- 12733281 TI - Metallic and inorganic mercury poisoning. PMID- 12733282 TI - Minor and major patients--what can be done? PMID- 12733283 TI - Paying the price for lifelong learning. PMID- 12733284 TI - Good practice should limit the spread of SARS in the UK. PMID- 12733285 TI - Are nurses losing sight of the patient? PMID- 12733286 TI - Family values. PMID- 12733287 TI - Complications associated with myocardial infarction. AB - The incidence of complications after acute myocardial infarction (MI) has been estimated to range from 14-95 per cent, with an overall one-month mortality of 30 per cent. Early treatment, as advocated by the National Service Framework for Heart Disease, has brought about some reduction in associated morbidity and mortality after MI. This article reviews the common complications associated with an acute MI, such as cardiogenic shock, pericarditis and heart failure. Nurses who are knowledgeable about potential complications should be able to detect early signs and symptoms, initiate emergency treatment, and prevent profound haemodynamic compromise occurring. PMID- 12733289 TI - Brief interventions for people who drink heavily. PMID- 12733288 TI - What you need to know about...pulmonary embolus. PMID- 12733291 TI - Sight loss in later life: a vision for health service intervention. AB - The care and support of older people with visual impairment is a neglected public policy issue, despite the growing numbers affected by sight loss in later life. A recent study of 400 people aged above 55 has identified problem areas and ways to improve practice and service delivery. Improvements will depend on better patient care at the time of diagnosis and more effective follow-up, with implications for closer liaison between health and social care practitioners. The article highlights three significant issues: the quality of patient care at time of diagnosis; difficulties experienced receiving and gaining access to outpatient follow-up; and the consequences of low levels of registration. PMID- 12733290 TI - Management of malignant spinal cord compression. AB - Malignant spinal cord compression is a relatively uncommon but serious condition that needs to be treated with urgency to prevent paralysis. Formal guidelines on procedures to be followed if malignant cord compression is suspected, and increased education of patients, nurses, and doctors, could help avoid some delays in treatment. PMID- 12733293 TI - Straight to the heart. PMID- 12733292 TI - Tackling the taboo. PMID- 12733294 TI - Could islet transplantation be a potential cure for diabetes? AB - Islet transplantation is a way of replacing the insulin-producing beta cells in patients whose own cells have been destroyed. Sue Swift and Steve White explain what islet transplantation involves and give a realistic summary of what can be expected as the clinical programme gets underway in the UK. PMID- 12733295 TI - The delivery strategy for the National Service Framework for Diabetes. AB - The National Service Framework for Diabetes: Delivery Strategy is now available. It aims to reduce variations in diabetes care in England. Mary MacKinnon outlines the implications for nurses working with people with diabetes. PMID- 12733297 TI - Obesity and diabetes. AB - Obesity increases the risk of developing type 2 diabetes. Jacqueline Cleator and John Wilding describe treatment strategies including diet, activity, pharmacotherapy and surgery. They argue that nurses need to gain a greater understanding of nutritional management and behavioural psychology in order to contribute effectively to treatment. PMID- 12733296 TI - Devices for insulin administration. AB - The drive to improve blood glucose control to prevent diabetic complications has resulted in many patients with type 2 diabetes, as well as those with type 1, requiring insulin therapy. Jillian Hill describes the devices now available to deliver insulin, which enable more people to manage their insulin administration independently. PMID- 12733298 TI - Management of type 1 diabetes and perioperative fasting. AB - Pre and postoperative fasting presents specific problems to patients with type 1 diabetes. Too much insulin can cause hypoglycaemia, whereas too little can lead to ketoacidosis. Simon Eaton explains how avoiding these complications requires use of appropriate treatment regimes and careful monitoring of glucose levels. PMID- 12733299 TI - Insulin and normal eating. AB - Sarah Bloor critiques a research paper about a randomised control trial, which examined the use of training in flexible, intensive insulin management to enable dietary freedom for people with type 1 diabetes. (Dose Adjustment for Normal Eating Study Group, 2002). PMID- 12733300 TI - Managing care in the community for patients with type 2 diabetes. AB - Jean Peters and colleagues researched the perceptions of 97 practice nurses and 69 diabetes specialist nurses regarding their current and future role in the management of people with type 2 diabetes in the community. Issues of concern that were identified included patients, resources, training and professional responsibilities. PMID- 12733301 TI - How to ... lead a team. PMID- 12733302 TI - [Is screening radiography for cancer of the lung still current?]. PMID- 12733303 TI - [Pasteur followers and medicine of the 20th century]. PMID- 12733304 TI - [Infections by Candida sp. in intensive care. Survey of French practices]. AB - OBJECTIVE: The isolation of Candida sp in nosocomial infections is on the increase and over the past 10 years many guidelines for "good" practices and recommendations have been published on the modalities for the management of systemic candidiasis. The aim of this paper was to assess the habits in the intensive care units in this domain in France. METHOD: A transversal survey on the habits was conducted from March to May 2001, using a questionnaire mailed to 200 intensive care units. RESULTS: One hundred eighty questionnaires (surgical reanimation: 12%, medical: 18%, medico-surgical: 70%) out of 200 (92.5%) were returned. The indirect diagnostic examinations: serology, search for antigenemia and PCR (Polymerase Chain Reaction) were never used in 21, 35 and 65% of cases. The systematic search for colonisation (a mean of 4 areas sampled) was conducted in all the patients by 19% of the investigators, in some patients by 53%, and never by 28%. An antifungal treatment was prescribed: in the presence of a positive haemoculture alone, once out of twice if the sample had been taken from a central catheter and in 2 cases out of 3 when the sample was peripheral. It was prescribed 6 times out of 10 after isolation of Candida sp following surgery or on needle aspiration of an intra-abdominal abscess, varyingly in the case of cadiduria, isolation of a Candida sp in a broncho-pulmonary sample or in abdominal draining and positive culture of a catheter, depending on the intensity of the colonisation, the severity of the clinical picture and the presence of factors of risk for Candida infection. It is still prescribed empirically depending on the same elements and the absence of explanation for worsening. When faced with candidemia in a non-neutropenic patient, a central catheter is not changed in 18% of cases. Depending on the microbiology, fluconazole is prescribed in: the identification of yeast without further precision (78% of cases), Candida sp without further precision (86% of cases), Candida non albicans without further precision (57% of cases), C. albicans (93% of cases), Candida non albicans other than C. krusei and C. glabrata (62% of cases), C. glabrata (36% of cases) with an increase in dose in 1 out of 2 cases. In the presence of C. glabrata or C. krusei, amphotericin B is the choice in respectively 51 and 75% of cases. To adapt the treatment. PMID- 12733305 TI - [Complications due to peripheral venous catheterization. Prospective study]. AB - OBJECTIVE: Peripheral venous catheter (PVC)-associated complications were prospectively evaluated in a 2 month-study performed in 3 different wards. METHODS: For each inserted PVC, the following complications were observed daily by an external investigator: tenderness, erythema, swelling or induration, palpable cord and purulence. PVC that were removed were systematically sent to the Microbiology department and analysed according to the semi-quantitative method described by Brun-Buisson et al. RESULTS: A total of 525 PVC (corresponding to 1,036 catheterisation-days) were included. Main clinical complications were erythema (22.1%), tenderness (21.9%), swelling or induration (20.9%), palpable cord (2.7%) and purulence (0.2%). Phlebitis, defined by 2 or more of the following signs: tenderness, erythema, swelling or induration and palpable cord, was observed in 22%. Catheter colonization (> or = 103 CFU/ml) occurred in 13%. Bacteria isolated from colonized catheters were coagulase negative staphylococci (88.1%), Staphylococcus aureus (7.1%) and Candida sp. (4.8%). Multivariate risk factor analysis showed that age > or = 55 y. (OR = 3.16, p = 0.003), insertion on articulation site (OR = 2.94, p = 0.01) or in jugular vein (OR = 8.18, p = 0.01) and > 72 hour-catheterisation (OR = 4.74, p = 0.0003) were significantly associated with PVC colonization. Risk factors for phlebitis were skin lesions (OR = 1.88, p < 0.016), active infection unrelated to PVC (OR = 2.8, p = 0.001), "poor quality" peripheral vein (OR = 2.46, p < 0.02) and > 72 hour-catherisation (OR = 2.38, p = 0.009). CONCLUSION: Complications associated with peripheral venous catheters are frequent but remain benign. They could probably be reduced by a systematic change every 72-96 hours as recommended by different guidelines. PMID- 12733306 TI - [A rare cause of exophthalmos, hydatid cyst of the orbit]. AB - INTRODUCTION: The orbital hydatid cyst is a rare (1 to 2%) localization of the Echinococcus granulosus parasite. OBSERVATIONS: An intra-orbital localization of hydatidosis was revealed in two patients by the occurrence of unilateral exophthalmia. After tumoral resection the diagnosis was confirmed by histological examination. COMMENTS: The principle clinical sign of an orbital hydatid cyst is exophthalmia. Imaging is essential for pre-operative diagnosis. Serology is insufficient and treatment is surgical. PMID- 12733308 TI - [Unusual discovery of hepato-pulmonary fusion during right congenital diaphragmatic hernia surgery]. PMID- 12733307 TI - [Inclusion myositis associated with primary biliary cirrhosis of the liver]. PMID- 12733309 TI - [Chronic sarcoid myositis]. PMID- 12733310 TI - [Livid streaks induced by bleaching creams]. PMID- 12733312 TI - [Pharmacologic innovations, the fluoroquinolone model]. AB - OBJECTIVES: Like all the disciplines involved in infectious diseases and antibiotic therapy, pharmacokinetics and pharmacodynamics (PK/PD) of antibiotics have evolved significantly. They include new investigation procedures like in vitro models and new animal models. There is a current trend towards better methods, newer definitions and improved quality of research in this particular field. DEFINITIONS: The current evolution of pharmacology of antibiotics is mainly characterized by the development of pharmacodynamics (PD) which offers the advantage of including microbiology data. New PD parameters have been defined. It is recommended to working groups involved in PK/PD research to compare their results and tend to a consensus on the methods to be used. CONCLUSION: Benefiting from such progress, preclinical phases of PK/PD studies of new molecules as well as revision of older antibiotics permit a rigorous and clinical approach to the use of antibiotics. PMID- 12733311 TI - [Botulism toxin, bioterrorist weapon]. AB - BOTULISM AND BIOWARFARE: Botulism is a severe neuro-paralysing infection due to a toxin produced by Clostridium botulinum. The use of the botulinum toxin for terrorist aims in the form of aerosols is a perfectly credible eventuality. The botulinum toxin is the most potent toxin known; it is easy to produce and can lead to massive destruction. DEPENDING ON THE CONTAMINATION: The clinical forms of botulism depend on the mode of contamination. Botulism through inhalation can only be the result of a deliberate act using an aerosol. The clinical symptomatology is identical to that of the other forms. PREVENTION: In the case of a bio-terrorist attack with an aerosol of botulinum toxin, the subjects exposed should be vaccinated as a prophylactic measure with trivalent antitoxin vaccine (types A, B and E). This vaccine must be administered as rapidly as possible in symptomatic patients. A single case of botulism acquired by inhalation corresponds to an act of terrorism. PMID- 12733313 TI - [Management of patients infected with HIV]. AB - HIV-VIRAL HEPATITIS CO-INFECTIONS: Several characteristics reveal an HIV-HBV co infection: high B viral replication, high percentage of patients exhibiting chronic B virus, high risk of cirrhosis, and possibility of B reactivation in severely immunodeficient patients. Regarding HIV-HCV co-infections, there is a greater risk of progression towards cirrhosis. However, anti-retroviral treatment appear to stall the progression of the C-virus hepatic disease. METABOLIC COMPLICATIONS WITH ANTI-RETROVIRAL TREATMENTS: Among the morphological lipodystrophic syndromes, the lipohypertrophic forms must be distinguished from the lipoatrophic forms. Substitution of some antiretroviral molecules is the first measure to be taken, but the results are difficult to assess; other current drug alternatives are unconvincing. The management of hypercholesterolemia and hypertriglyceridemia observed during treatment with antiretrovirals is debatable, and efficacy is not always clearly demonstrated. Regarding hyperlactatemia, potentially the most severe complication of mitochondrial toxicity of antiretroviral treatment, which requires suspension of the nucleoside analogs in severe or moderate symptomatic forms, and simple surveillance and continuation of the treatment in the mild or moderate asymptomatic forms. VIROLOGICAL FAILURE: There are three options possible: continue the same treatment, change it or stop it. Efficacy should be assessed on CD4 and the variations in viral load, rather than on the absolute value of the viral load. IN POOR RESSOURCE SETTINGS: Only a minority of patients has access to antiretroviral treatments. Efforts must be made to continue to search for other forms of management: community measures for prevention and early screening, psychological and nutritional support, prophylactic and treatment strategies for infections or opportunist diseases. PMID- 12733314 TI - [Some current aspects of drug resistance by staphylococcus]. PMID- 12733316 TI - [Nystagmographic findings in pontocerebellar angle tumors]. AB - OBJECTIVE: To determine the diagnostic and prognostic value of vestibular function tests in cerebellopontine angle tumors METHODS: Analysis of preoperative nystagmographic findings in 29 patients affected of cerebellopontine angle tumors and its possible correlation with anatomical and progressive patterns RESULTS: Tumor size and growth pattern were statistically significantly related to auditory and vestibular functions and to central findings on nystagmography. Age, gender, tumor size and central findings on nystagmography did not show influence upon postoperative dysbalance. However, alterations related with adaptation of the vestibular system (positional nystagmus and asymmetry on rotational tests) correlated significantly with postsurgical dysbalance. CONCLUSIONS: The tests for vestibular function reflect a the characteristics of growth of the cerebellopontine angle tumors and can help to identify the patients susceptible of prolonged postoperative dysbalance. PMID- 12733315 TI - [Ontogenic peculiarities of the human tympanic ossicular chain]. AB - We have studied the development of the tympanic ossicles in 40 embryo-foetal human series aged between 32 days (6 mm) and newborn. Once performed the measurements to date chronologically embryos and foetuses, we did a meticulous dissection of temporal bones. After fix in 10% formol, decalcified with 2% nitric acid, embedded in Paraplast, sectioned in a sequence of 7 mm, and stained with Martin's trichrome. The tympanic ossicles are developed in the mesenchyme of the two first pharyngeal archs. The head of the malleus, the body and the short limb of the incus arise from the first arch while the handle of the malleus, the long limb of the incus and the mass of the stapes arise from the second arch. The vestibular side of the stapedial footplate develops in the otic capsule. The tympanic ossicles develop from endochondral ossification, while anterior process of the malleus has the membranous ossification. In their ontogenia 6 stages are observed. First stage, the formation of their sketch by mesenchimal condensation, in the second stage, "pre-cartilaginous", the cells of the primordia are differentiated into condroblasts, in the third stage "cartilaginous" the ossicles show a cartilaginous structure, in the forth stage the primary ossification centers are developed, in the fifth stage the ossicles arise in the periostic annulus and inside the endochondral bone, and in the last stage the osseous tissue grows until it acquires a compact osseous structure. PMID- 12733317 TI - [Genetic alterations in sinonasal adenocarcinoma in wood workers studies with comparative genomic hybridization]. AB - Eleven wood-workers with sinonasal adenocarcinoma were analyzed by comparative genomic hybridization. This technique serves as a screening test for regions of copy number changes in tumor genomes. We have applied the technique to map DNA gains and losses in 9 cases of formalin-fixed, paraffin-embedded tumors and 2 cases of frozen tumors. Gains were found most frequently than losses. Whole arm chromosomic gains were detected in high frequencies at 8q, 7q, 12q and 18p and losses at 18q, 8p, 10q, 5q, 14q and 17p. The segment most frequently amplified was 18p11.1-q11 (45%), even though others like 7q21-22 (18%) were related with lower survival rates. This analysis allows us to know for the first time the chromosomic aberrations that occur and may play an important role in sinonasal adenocarcinoma. In the future, comparative genomic hybridization could be used in the woodworkers with long exposition to wood dust to detect initial genetic aberrations and obtain an early treatment and diagnosis of the disease. PMID- 12733318 TI - [Lymphoma of the nasal cavity, clinical and anatomo-pathologic considerations]. AB - Six cases of high-grade lymphoma affecting the nasal cavity are presented. Their main symptoms were nasal obstruction, epistaxis and rhinorrea. Clinical exploration showed an ulcerated lesion in four patients (all of them T/NK cell lymphomas) and an exofitic tumour in the other two (B cell lymphomas). Pathology was diagnostic for high grade lymphoma in all the six cases, immunohistochemical studies revealed that the atypical cells were T/NK in 4 cases and B in the other two cases. Genotypic analysis was done looking for rearrangement of the genes for Ig H or T-cell receptors. In situ hybridisation was done to detect Epstein-Barr virus RNA. Malignant lymphomas arising in the nasal cavity are unusual, its major prognostic factor is the clinical stage at presentation. To perform an early diagnosis this neoplasia must be suspected in the presence of an exofitic tumour (B cell lymphoma) or necrotic and ulcerated lesion (T/NK cell lymphoma). PMID- 12733319 TI - [Treatment of T1N0 glottis carcinoma with radiotherapy. Results at our center and review of the literature]. AB - OBJECTIVE: To evaluate the results of radiotherapy in the treatment of patients with T1N0 glottic squamous cell carcinoma. MATERIAL AND METHODS: Retrospective study of a cohort of 338 patients with T1N0 glottic carcinoma treated with radiotherapy at our institution between 1985-1997. A review of the literature published during the last ten years was carried out. RESULTS: Local control with radiotherapy in our patients was 82%, reaching 97% when salvage surgery was included. The local control with radiotherapy in most of the published series ranges between 81.90%. CONCLUSIONS: Treatment with radiotherapy achieves local control in early glottic carcinomas (T1N0) in 80-90% of cases. In our centre such treatment achieved local control in 82% of cases. PMID- 12733320 TI - [Spread channels of anterior commissure cancer: clinico-pathological study and surgical implications]. AB - The aim of the present study is to describe the spread channels of the anterior commisure cancer, its clinical significance and its surgical implications. PATIENTS AND METHODS: A prospective study was performed at the ENT Department of the Complutense University in about 31 patients with anterior commisure carcinoma wo underwent surgery between 1994/97. Specimens were processes with Whole-Organ Serial Sections. RESULTS: Patients were differenciated into two groups: 1) Patients with glottic tumors, with a good cord mobility and no invasive tumor in commisure region (18 patients); In these patients, conservative surgery was possible; 2) Another group (13 patients) with aggressive lesions (80% with cartilage involvement), vocal cord fixation and transglottic lesions. In these group, conservative surgery was contraindicated. CONCLUSIONS: This report emphasizes the importance of laryngoscopy in surgical technique of anterior commisure cancer. PMID- 12733321 TI - [Oromandibular reconstruction with free peroneal flap and osseointegrated implants]. AB - Free fibula flaps have proved to be one of the most versatile for oromandibular reconstruction due to the available length of bone and the possibility of incorporating a long skin paddle to cover intraoral soft tissues. The use of a osseointegrated dental implants is an important technique for the oral rehabilitation of these patients. Osseointegrated implants provide the most rigid prosthetic stabilization available to withstand masticatory forces. These implants can be placed immediately or in second time procedure. In our case, implantation in the fibula free flap is done after 6-9 months because of the large amount of osteosynthesis material required for the fixation of the flap. Four or six months later, when osseointegration has taken place, the implants are loaded with a dental rehabilitation. We analize 10 cases of mandibular reconstruction with fibula free flap and their aesthetic and functional rehabilitation with osseointegrated implants with a 2 year follow up. Forty-six dental implants were placed developing all of them but one a correct osseointegration. All these patients recovered masticatory function and underwent a considerable improvement in labial competence, salivary continence, speech articulation and facial harmony. PMID- 12733322 TI - [Epidermoid cyst of the parapharyngeal space]. AB - Epidermal cysts of parapharyngeal location are very unusual tumors, with a benign character, though when they reach a great size, adjacent zones and vascular and nervous structures of great importance can be affected. We are going to describe the clinical aspects of this case, the instructions and the surgical approach used: the Fabre inferior and internal jaw approach of the lateropharingeal space and infrasphenotemporal fossa, which avoids the aesthetics and functional aftereffects of other more common approaches. PMID- 12733323 TI - [Tongue necrosis as onset of temporal arteritis]. AB - The Horton's disease is a generalized vascular affectation, characterized by a giant cell arteritis. Medium and small caliber arteries are the most frequently affected, mainly of the craneal vascular territory, in particular the collateral branches of the external carotid, and artery in special the superficial temporal artery. We present a case of lingual necrosis as debut of the Horton's disease that has been diagnosed and treated in our ENT Department. PMID- 12733324 TI - [Variations of salivary dismutase superoxide in tonsillar infection]. AB - The antioxidant effect of superoxido dismutase in saliva was measured in children bearing of tonsillar hypertrophy, recidivant tonsillitis or peritonsillar abscess. These levels were compared to those detected on tonsillar tissue obtained from tonsillectomy (p < 0.001). Although salivary SOD concentration in children with tonsillar infection was higher than hypertrophy, there was not a significative correlation to tonsillar value of the enzyme (R2 = 0.2276), so we can not accept a predictive value for salivary SOD of tonsillar suffering and, eventually, of tonsillectomy. PMID- 12733326 TI - [Dietary intake assessments: for who? why?]. AB - The use of dietary intake assessment and questionnaires in clinical practice is largely debated, particularly for obese patients. The time needed to carry out them is very long, and the results are little or not at all workable due to the underreporting which may represent up to 50% of intakes for some patients. The usual methods and tools to evaluate dietary intakes are presented, they should not be used to calculate energy intake, but they permit to initiate a dialogue with the patient on his/her dietary pattern. The analysis of the dietary pattern is the first step in diet prescription and is essential to weight management and follow-up. This analysis must consider 3 important components: type of food usually eaten and preferred, the circumstances and environment of food intakes and meals, possible disorders of eating behaviour. Some results on food consumption in adults and children from the Fleurbaix Laventie Ville Sante Study are presented. PMID- 12733325 TI - [To burn or to store]. AB - Energy exists as organic molecules and heat in living organisms. In adult mammals, body weight and fat content remain unchanged if energy intake is strictly equivalent to energy expenditure. In other words, regulation of body weight requires energy of foods to be entirely dissipated as heat. Imbalance between ingested energy and thermogenesis induces obesity or thinness. Alterations of food intake or energy expenditure represent the two causes of body weight disturbance. It is accepted that individuals differ in food efficiency i.e. ability to metabolize foods and store fat or totally burn nutrients. Mechanisms of food efficiency and futile cycles are presented. I started my research work analysing thermogenic mechanism in brown adipose tissue. Actually, in addition to white adipose tissue which is the major type of adipose tissue, mammals own another type of adipose tissue referred to as brown adipose tissue. This later tissue is an activatable thermogenic organ which oxidizes fatty acids and releases heat in blood stream. Brown fat is activated during exposure to the cold (in rodents), at birth, and during arousal in hibernators. My initial work helped to characterize a mitochondrial protein named uncoupling protein or UCP which is responsible for activation of fatty acid oxidation and heat production in brown adipocytes. Actually, in most cells, fifty per cent of oxidation energy is recovered as ATP in mitochondria through the process of coupling of respiration to ADP phosphorylation. In contrast to mitochondria of most tissues, brown adipocyte mitochondria can escape the obligatorily coupling of respiration and waste almost ninety per cent of respiration energy as thermogenesis. UCP characterization and its molecular cloning as well as antibodies obtention were used to better understand cellular thermogenesis. Brown adipocytes were identified in babies and adult patients with pheochromocytoma. More recently, research on the brown fat UCP helped us to identify UCP2, a UCP homolog present in most human and animal tissues. A family of UCPs exist in animals and plants. These UCPs may function as mitochondrial uncouplers. However, the ancient function of the UCPs may be rather associated to adaptation to oxygen and control of free radicals than to thermogenesis. Further studies of UCPs will improve the knowledge of mitochondrial metabolism and substrate oxidation. In other respects, analysis of molecular mechanisms controlling respiration uncoupling may contribute to new strategies of treatment of metabolic disorders such as obesity. PMID- 12733327 TI - [Energy expenditure: method employed]. AB - Energy expenditure (or energy needs) corresponds to the energy required for the body to work under all circumstances. This is the energy that should be taken (with food) to maintain energy balance, i.e. the constancy of body weight. Energy is expended at rest (referred to as basal or resting metabolic rate), when food is consumed (diet induced thermogenesis) and with physical activity. In obese people, energy expenditure is increased because of the proportion between body mass and energy expenditure. There is no definite proof that there is a metabolic defect in obese persons by which energy expenditure would be lower than expected for body mass. Hence obesity can only be the result of a positive energy balance. Energy expenditure can be measured by indirect calorimetry or estimated with equations. These equations use factors as weight, age, gender and sometime height. These equations are generally valid although there is a need for more validation studies. This paper also describes in which clinical circumstances the measurement or the estimate of energy expenditure are useful for the care of obese patients. PMID- 12733328 TI - [How do you really know if the obese patient has sleep apnea?]. AB - Obesity is known to predispose to obstructive sleep apnea (OSA), a condition characterized by repeated episodes of apnea or hypopnea during sleep, due to the interruption of airflow through the nose and mouth. These episodes lead to the fragmentation of sleep and to decrease in oxyhaemoglobin saturation. Patients with massive obesity, with or without daytime hypersomnolence should be systematically screened for OSA, because many of them appear to be asymptomatic and unaware of their breathing abnormalities during sleep. Polysomnography (PSG) in an attended hospital laboratory setting is the gold standard for the diagnosis of OSA. However portable recording devices can be used for screening with good sensibility and specificity, and even for diagnosis when the apnea-hypopnea index is high. However the final diagnosis can only be carried out in a sleep laboratory using PSG by highly-qualified personnel, because of the limitations of the portable recording device. There is a strong association between OSA and the risk of traffic accidents. It has been established that OSA affects quality of life. There is also increasing evidence that OSA is an independent risk factor for cardio-vascular diseases. This has been successfully demonstrated for hypertension by prospective studies. But the evidence remains weak for myocardial infarction, stroke or mortality. Treating OSA with continuous positive airway pressure (CPAP) is the treatment of choice. CPAP improves quality of life, driving simulator performance, blood pressure and sleepiness, as demonstrated by randomised placebo controlled trials. The majority of obese OSA patients are currently not being offered diagnosis testing and treatment. It's a real challenge due to the epidemic increase of obesity prevalence. Portable recording devices could be available outside the sleep laboratory in nutrition department, where morbid obesity is treated. This emphasizes the need for a real collaboration between these departments and sleep experts. PMID- 12733329 TI - [Are diets fattening?]. AB - A diet always induces a weight loss in the short term. The loss does not depend on the diet composition but rather on the caloric deficit. However, a drastic diet often induces binge eating disorders and can lead to a weight gain in the long term. A cognitive-behavioural-nutritional approach allows a lasting weight loss. Results are much better in the long term with low fat diets. The benefits of a weight loss, even a little one, improves greatly morbidity and mortality at 10 years. PMID- 12733330 TI - Diagnosis of Helicobacter pylori infection in patients with upper gastrointestinal bleeding due to peptic ulcer. PMID- 12733331 TI - The effectiveness of (IgG-ELISA) serology as an alternative diagnostic method for detecting Helicobacter pylori infection in patients with gastro-intestinal bleeding due to gastro-duodenal ulcer. AB - OBJECTIVES: To establish the sensitivity, specificity, positive predictive value and negative predictive value of serology (IgG ELISA) as an alternative diagnostic method for Helicobacter pylori infection in patients with gastro duodenal peptic ulcer and digestive hemorrhage. The diagnosis of Helicobacter pylori infection in these patients is difficult due to the low sensitivity of invasive tests and the need to discontinue treatment with proton pump inhibitors to perform a breath test with urea 13C or the detection of Helicobacter pylori antigens in feces. PATIENTS AND METHODS: We included 214 patients (164 men and 50 women) with an average age of 58 +/- 15 years, who were admitted to hospital due to upper gastro-intestinal bleeding caused by a gastro-duodenal peptic ulcer. The presence of Helicobacter pylori was established by means of gastric biopsy (fast urease test histology and/or culture) and a breath test with 13C-labeled urea. Serology was performed with the ELISA method (Pyloriset EIA-G by Orion Diagnostica). Positive Helicobacter pylori infection was accepted with any positive invasive method or breath test, and no infection was established if all invasive tests performed and the breath test with 13C-labeled urea were negative. We calculated the sensitivity, specificity, positive predictive value and negative predictive value of serology in the global series and in different subgroups of patients according to age (> 60 and < 40 years), recent exposure to non-steroidal anti-inflammatory drugs, type of endoscopic wound and history of gastro-duodenal peptic ulcer. RESULTS: 192 patients (89.7%) showed infection due to Helicobacter pylori. In the global series (n = 214) we obtained a sensitivity, specificity, positive predictive value and negative predictive value of 87.5, 54.5, 94.3 and 33.3%, respectively. Specificity was greater in the group not exposed to non-steroidal anti-inflammatory drugs (n = 110) as compared to the exposed group (n = 104), in the < 40 year old group (n = 28) with respect to the > 60 years group (n = 105), in the duodenal ulcer group (n = 141) with respect to the gastric ulcer group (n = 59), and in the group with a history of gastro duodenal peptic ulcers (n = 92) as compared to the group without any of these past events (n = 122); nevertheless, no significant statistics were reached. CONCLUSIONS: IgG (ELISA) serology shows low specificity and a low negative predictive value in the diagnosis of Helicobacter pylori infection in patients with gastro-intestinal bleeding due to gastro-duodenal peptic ulcer. The diagnostic value of serology did not improve significantly when age, recent exposure to non-steroidal anti-inflammatory drugs, type of endoscopic wound or history of gastro-duodenal peptic ulcer was taken into consideration. We may consider that serology is not a good diagnostic method for the detection of Helicobacter pylori in patients with digestive hemorrhage caused by gastro duodenal ulcer. PMID- 12733332 TI - Protective effect of vagotomy on the gastric mucosa in a stress model in rats. AB - INTRODUCTION: The use of vagotomy is classically based on its inhibiting effects on acid secretion. Vagotomy induces both cellular and endocrine changes that may be involved in protective actions. Our aim was to study morphologic changes induced by vagotomy on the gastric mucosa and their relation to stress protection in the short, medium and long term. MATERIAL AND METHODS: An immobilization and cold stress model was used with 80 Wistar rats divided into two groups--control (with and without stress) and vagotomy (at 7, 30 and 120 days). Changes induced in the gastric mucosa by stress were studied with and without vagotomy, as well as relationship between these changes and the intended protective action. RESULTS: Bleeding showed a very significant relation to stress (p < 0.0001). Bleeding incidence exhibited a significant difference between vagotomised and non vagotomised rats (p < 0.0001) in the short, medium and long term (vagotomy was protective against stress). Regeneration signs related significantly to vagotomy (p < 0.0001) but not stress (p = 0.208). However, no significant relationship was found between the protective action and the presence of regeneration signs (p = 1). CONCLUSIONS: Vagotomy has tropic effects on the gastric mucosa and is protective against stress. This protective action is maintained in the short, medium and long term. However, these changes are not sufficient to explain protection. An adaptation phenomenon mediated by hormonal and peptidic factors may be involved in this action. PMID- 12733333 TI - Adaptation and validation of a gastroesophageal reflux questionnaire for use on a Spanish population. AB - OBJECTIVE: To produce and validate for telephone use in Spain a cross-cultural adaptation of the gastroesophageal reflux questionnaire (GERQ) developed by Locke et al. MATERIAL AND METHOD: The Spanish version of the questionnaire was developed through translation, assessment of comprehensibility, back translation, and appraisal of applicability. Subsequently, reproducibility was measured by a test-retest procedure in 125 patients, and concurrent validity was evaluated by comparing self-reported questionnaire data against a gastroenterologist's findings from clinical interviews with 50 patients. RESULTS: The Spanish version showed itself to be easily understandable and widely applicable. Reproducibility for the two cardinal symptoms of gastroesophageal reflux disease (heartburn and acid regurgitation) as measured by the kappa index (95% confidence interval), was 0.85 (0.73-0.97) and 0.81 (0.70-0.92), respectively. With respect to the process of concurrent validation, kappa values (95% confidence interval) for heartburn and acid regurgitation were 0.85 (0.68-1) and 0.91 (0.79-1), respectively. Median kappa for all questions on the questionnaire was 0.75 (range: 0.42 a 1) in the reproducibility analysis and 0.83 (range: 0.44 a 1) in the concurrent validity analysis. CONCLUSIONS: The reproducibility and concurrent validity of the Spanish version of the GERQ is excellent and comparable to that of the original English version. PMID- 12733334 TI - Antidepressant-induced hepatotoxicity. PMID- 12733335 TI - Giant diverticulum of the sigmoid colon. AB - The giant colonic diverticulum is a very rare clinical entity usually located in the sigmoid colon of elderly patients. A case of an 87-year-old woman recently treated in our hospital is reported hereinafter. The patient was non-surgically treated due to her advanced age and high surgical risk. PMID- 12733337 TI - [Amebic liver abscess. Surgical treatment]. PMID- 12733336 TI - [Cytomegalovirus colitis and recto-vaginal fistula in an immuno-competent woman]. PMID- 12733338 TI - [Choledochocele associated to neoplasm]. PMID- 12733339 TI - [Authors reply: primary hepatic abscess caused by Actinomyces meyeri]. PMID- 12733340 TI - [The influence of type I collagen on the cell behavior of human embryonic periosteous osteoblasts]. AB - OBJECTIVE: To study the influence of type I collagen (COL I) on the cell behavior of human periosteous osteoblasts (OB) and the application of type I collagen in constructing bioactive artifical bone. METHODS: OB were cultured on dishes coated with bovine type I collagen in different final concentrations. The cell adhesion was examined by the methods of cell count, the proliferation of OB was studied by 3H-TdR, and the osteoblastic ability was assessed by the synthesis of collagen, osteocalcin and alkaline phosphatase (ALP). RESULTS: OB cultured on type I collagen layer had the following characteristics: 1. The amounts of adhesive cells were maximal top in 25 micrograms/ml final concentration; 2. The proliferation of OB was decreased above 12.5 micrograms/ml final concentration (P < 0.05); 3. The synthesis of type I collagen was reduced slightly (above 25 micrograms/ml, P < 0.05); 4. The secretion of osteocalcin was increased markedly (above 6.25 micrograms/ml, P < 0.05, which reached maximally in 25 micrograms/ml); 5. The ALP activity was also increased (above 12.5 micrograms/ml, P < 0.05). CONCLUSION: Type I collagen promotes the expression of osteoblastic phenotype and cell adhesion. When the scaffold materials for bone tissue engineering are coated with type I collagen, the osteogenesis of OB is enhanced to accelerate the transformation course from artificial bone to biological bone, the best final concentration is 25 micrograms/ml. PMID- 12733341 TI - [Study on the construction of DYS385 allelic ladder and the genotype distribution in two populations]. AB - OBJECTIVE: We have designed a new method to produce standard DYS385 allelic ladder in order to solve the problem of the accuracy and standardization of STR PCR typing in forensic practice. METHODS: Nine different PCR amplified DYS385 allelic fragments were isolated from the gel, eluted into the distilled water and reamplified by PCR. The purified allelic fragments were then blunt-end subcloned individually into the pUC plasmid vectors and transfected into competent E. coli DH5 alpha cells. The sequencing results confirmed that the size and the constructure of the inserts were correct. The recombinant plasmids DNA with 9 inserts were then used as templates for reamplification to generate DYS385 standard ladder. RESULTS: A large quantity of standard DYS385 allelic ladder was obtained, with which the genetic polymorphisms of DYS385 locus both in Chinese Han and German populations were studied. CONCLUSION: The STR standard ladder constructed by this method has high value in the forensic application, and the DYS385 locus is robust for forensic analysis. PMID- 12733342 TI - [Biochemical remodeling of myocardial collagen and its relation to cardiac renin angiotensin system in rats after myocardial infarction]. AB - OBJECTIVE: This study aimed at the biochemical remodeling of myocardial collagen and its relation to cardiac renin-angiotensin system (RAS) after myocardial infarction(MI). METHODS: The ventricular and plasma angiotensin II (Ang II) content, total collagen content, and collagen concentration in left ventricle(LV) and right ventricle(RV) of MI and sham-operated control(SOC) rats on 3, 15 and 42 days after operation were measured dynamically by means of radioimmunoassay etc. RESULTS: The plasma Ang II content increased only in MI rats on day 3. The Ang II content in LV of the MI group increased markedly on day 3 after MI as compared with that of the SOC group. It sustained at this higher level on the 15th and 42th days, and the similar changes were observed in RV as well. The biochemical remodeling of collagen in LV and RV occurred after MI and the remodeling of collagen showed similar patterns with its Ang II content, except that collagen remodeling in RV was delayed than the change of Ang II content. CONCLUSION: The cardiac RAS was obviously activated after MI. Ventricular Ang II might play an important role in the biochemical remodeling of myocardial collagen, and its role was independent of circulating RAS. PMID- 12733343 TI - [Effect of tetrandrine on expression of bFGF in lung tissue of rat with chronic hypoxic pulmonary hypertension]. AB - OBJECTIVE: To evaluate effect of tetrandrine (Tet) on chronic hypoxic pulmonary by observing expression of bFGF in lung tissue of rat with chronic hypoxic pulmonary hypertension. METHODS: 30 rats were divided into 3 groups: hypoxia group, hypoxia + Tet group and control group. On the 21st day of hypoxia, the hemodynamic parameters were measured and the intra acinar pulmonary arteries (IAPA) changes were studied under light microscopes. Immunohistochemical staining with monoclonal antibody against human recombinant bFGF was performed in the paraffin section of rat lung. RESULTS: Tetrandrine could reduce pulmonary artery mean pressure (mPAP) and pulmonary vascular resistance (PVR), (P < 0.01), decrease the expression of bFGF in wall of IAPA, and inhibit hypoxic structural remodeling of IAPA. CONCLUSION: Tetrandrine could decrease the expression of bFGF in wall of IAPA and in that connection to inhibit remodeling of IAPA and pulmonary hypertension. PMID- 12733344 TI - [Expression of pyruvate oxidase gene sopox from Streptococcus sanguis in E. coli]. AB - OBJECTIVE: To reconstruct the expression plasmid of Sopox gene for further understanding the regulation of its expression. METHODS: Sopox was recombined with expression vector pBV220 and the expression of Sopox in E. coli JM105 was observed after transformation. RESULTS: pBV220/Sopox/JM105 expressed a protein with molecular weight of 65 kd on SDS-PAGE after induction, and the expression reached the maximal amount with induction at 42 degrees C for 4 hours. CONCLUSION: Sopox was successfully cloned into pBV220 and expressed in E. coli JM105. PMID- 12733345 TI - [The stereocontrolled synthesis of 1-acetylenyl-3-tert-butyldiphenylsilyloxy-exo bicyclo [3. 1. 0] hexane]. AB - OBJECTIVE: In order to find out more effective method for the synthesis of the A ring synthon, (-)-1-acetylenyl-3-tert-butyldiphenylsily-loxy-exo-bicyclo [3. 1. 0] hexane (3), for 19-nor-1 alpha, 25-dihydroxyvitamin D3(2)with commercial value. METHODS: We synthesized the A-ring synthon (+/-)(3) via nine steps with good yield starting from 3-cyclopentenyl alcohol(4). RESULTS: The spectral data of compound (+/-)(3) is the same as the spectral data of (-)(3) which reported preciously by us. CONCLUSION: The method is an effective method for synthesis of the A-ring synthon (3). PMID- 12733346 TI - [Effects of transforming growth factor beta 1 on the expression of integrin beta 3 and the activity of focal adhesion kinase on human vascular endothelial cell]. AB - OBJECTIVE: To observe the effects of transforming growth factors beta 1 (TGF beta 1) on the expression of integrin beta 3 and the activity of focal adhesion kinase (FAK). METHODS: This study was performed on cultured human endothelial cells (EC) by using cell-ELISA and immunoprecipitation-tyrosine kinase assay respectively. RESULTS: Under the stimulus of TGF beta 1, there a dose-dependent increase in the expression of integrin beta 3 chain in the surface of EC. And after the cultured EC were treated with 5 ng/ml or 10 ng/ml TGF beta 1 for 6 or 24 hours, the FAK activity in EC increased significantly as compared with the control group (P < 0.05). CONCLUSION: The expression of integrin beta 3 and the activity of FAK on EC were regulated by TGF beta 1, and this regulation may be important in cell adherence, angiogenesis, and in the pathophysiology of atherosclerosis. PMID- 12733347 TI - [Characterization of cefoperazone resistance gene on plasmid pFC in E. coli HX88108]. AB - OBJECTIVE: To investigate the characterization of cefoperazone resistance gene (CPZr) on plasmid pFC in E. coli HX88108 and inquire into the mechanism of resistance to CPZ at the molecular level. METHODS: E. coli HX88108 strain which demonstrated high-level resistance to cefoperazone (MIC, > 512 micrograms/ml) was isolated from a severely infected patient in 1988. Five plasmids coexisting in the strain were designated pFC, pFT1; pFT2, pFT3 and pFX, respectively. Four plasmids except pFX conferred CPZ resistance. Cefoperazone resistance gene (CPZr) has been cloned from plasmid pFC. beta-lactamase assays with Nitrocefin were performed. RESULTS: The expression product of CPZr was beta-lactamase. The high level beta-lactamase enzymatic activities against cephaloridine of CPZr transformants which were detected spectrophotometrically at 260 nm wave length demonstrated high level similarities to that of pFC. MICs of 18 antibiotics were determined according to a guideline of NCCLS by broth dilution method. CPZr transformants showed moderate level resistance to ampicillin, cefazolin, cefazolin, cefamandole and CPZ (MIC, 64 micrograms/ml). Meanwhile, susceptibility testing results demonstrated that the level of resistance to CPZ of pFC transformant in this study (MIC, 64 micrograms/ml) was much lower than that in 1988 (MIC, > 512 micrograms/ml) and resistance to nofloxacin and aminoglycosides was not observed. Induction experiment and temperature-sensitive mutation of CPZ resistance were performed. CPZr colonal strains revealed the higher-level of resistance to CPZ (MIC, 512 micrograms/ml) due to antibiotic CPZ induction rather than temperature sensitive mutation. CONCLUSION: This observation suggests that resistance to antibiotics encoded by plasmid might have been lower or lost under no antibiotic stress in a certain period, but higher under heavy stress. PMID- 12733348 TI - [Effects of estradiol and isoflavoid on the expression of adhesion molecules on neutrophils]. AB - OBJECTIVE: To elucidate the effects of estradiol and isoflavoid on the expression of adhesion molecules on neutrophils. METHODS: Neutrophils of healthy subjects and ischemic stroke patients were treated with tumor necrosis factor alpha (TNF alpha), in the presence or absence of different concentrations of isoflavoid (WZ1, WZ2) and estradiol (WZ3, WZ4). Flow cytometry was used to detect the expression of CD18 and CD62L on neutrophil surface. RESULTS: 1. 10 ng/ml TNF alpha could activate neutrophils of healthy subjects; it increased the expression of CD18 by 10% on neutrophil surface and shed CD62L from the surface as shown by a 15% decrease of the fluorescence intensity and a 30% decrease of the percentage of positive cell. 2. Isoflavoid (WZ1, WZ2) had no significant effect on the expression of CD18 and CD62L on neutrophils. 3. Pretreatment of neutrophils with estradiol (WZ3, WZ4) could inhibit the activation of neutrophils by TNF alpha, which decreased the fluorescence intensity of CD18 by 8%, increased the fluorescence intensity of CD62L by 15% and increased the percentage of CD62L positive cell by 20%. 4. TNF alpha could activate the neutrophils of ischemic stroke patients strikingly; it increased the fluorescence intensity of CD18 by 20% and decreased the fluorescence intensity and percentage of positive cell of CD62L by 30%, and there was a significant difference when the patients were compared with the healthy subjects. Estradiol had the same effect on the expression of CD18 and CD62L as on those of healthy subjects. CONCLUSION: 1. TNF alpha is a strong activator of neutrophils; it plays an important role in the occurrence and development of ischemic stroke. 2. Isoflavoid has no obvious effect on the expression of adhesion molecules on neutrophils, so its role in protecting the cardiovascular system may come into play not by the way of affecting adhesion molecules expression. 3. Pretreatment of neutrophils with estradiol could protect them from activation by TNF alpha, thus decreasing the expression of adhesion molecules, the adhesion of neutrophils -endothelial cells, and hence the risk of ischemic stroke. However, in case that the neutrophils have been activated by TNF alpha, estradiol has no effect on the expression of adhesion molecules; this implies that estradiol possibly has no valid anti adhesion therapeutic effect on ischemic stroke. PMID- 12733349 TI - [Effect on proliferation and apoptosis of human lingual carcinoma cells cotransfected by bax and p53 genes]. AB - OBJECTIVE: To investigate the effects on proliferation and apoptosis of human cancerous cells cotransfected by bax apoptosis-inducing gene and p53 tumor suppressor gene. METHODS: The chimeric gene pSV-CIP-bax-CAT was constructed in which bax gene was flanked upstream by a 217 bp fragment(+822(-)+1093) of the first intron and a 317 bp promoter fragment of human a 1(I) collagen gene. Human lingual carcinoma cell line Tca 8113 (LCC) in culture was respectively transfected with pSV-CIP-bax-CAT, and cotransfected with pSV-p53-CAT and pSV-CIP bax-CAT by using the transfection reagent DOSPER. RESULTS: Immuno-slot blot and ELISA demonstrated that the expression of bax and p53 genes increased remarkably in the transfected and cotransfected LCC, compared with the controls (LCC transfected with pSV-CIP-CAT and LCC). MTT colorimetric assay, TUNEL fluorenscence microscopy and flow cytometry showed that foreign bax or p53 gene inhibited the LCC growth and induced apoptosis (23.9% and 26.1% of inhibitory rate by bax gene and p53 plus bax genes respectively). CONCLUSION: The ectopic expression of the bax gene in the LCC promoted by the cis-acting elements of human a1 (I)collagen gene has obviously synergistic effect on the proliferation and apoptosis of the LCC contransfected with p53 gene plus bax gene for 48 hours. PMID- 12733350 TI - [Scaling the SF-36 in a Chinese population]. AB - OBJECTIVE: To facilitate the cross-cultural comparison, we employed the principles of fuzzy mathematics in scaling the SF-36 in a Chinese population. METHODS: We stratified an urban population according to the age, sex, and occupation, and selected 650 people to participate in the study. A total of 646 valid questionnaires were returned. In the 8 domains of the SF-36, the interval estimation of the scores of different alternative answers of 6 domains which have more than 2 alternative answers were conducted. RESULTS: The fuzzy degrees of most interval estimations were less than 0.5, except 4 estimations in the domain "physical functions". The ordering of the scores of 8 domains was more similar to the original assumption by our scaling criteria than by the US scaling criteria. CONCLUSION: The scaling criteria set in this study is more suitable for use in Chinese population and we suggest that the alternative answers of the domain "physical functions" be simplified to "yes" and "no". PMID- 12733351 TI - [Feasibility of using short form 36 in Chinese population]. AB - OBJECTIVE: To explore the feasibility of using the SF-36 in Chinese population. METHODS: We tested the reliability and validity of the SF-36 in 1603 rural residents, 646 urban residents, 70 high school girls, 122 patients with osteoporosis, and 150 stroke patients. RESULTS: The reliability and validity of the SF-36 in general were acceptable, however, the reliability and validity varied among different domains and in different populations. The reliability and validity of the domains "limitations in physical activities", "limitations in usual role activities because of physical health problems", "bodily pain", and "general health perception" were better than those of the domains "vitality", "limitations in social activities", "limitations in usual role activities because of emotional problems", and "mental health". CONCLUSION: The results indicate that we can use the SF-36 in Chinese population, but we have to pay attentions to the variance of the reliability and validity in different populations, especially for the interpretation of the results of the domains related to mental and social functions. PMID- 12733352 TI - [The norms of SF-36 scale scores in urban and rural residents of Sichuan province]. AB - OBJECTIVE: To develop the norms of SF-36 in Sichuan, China. METHODS: A total of 1603 rural residents and 646 urban residents were surveyed. Both the US scaling criteria and China scaling criteria were used to score the 8 domains of the SF-36 and compared with the norms of Hong Kong and US populations. RESULTS: The scores of the 8 domains of the SF-36 using China scaling criteria were highly correlated with those using the US scaling criteria, with correlation coefficients greater than 0.98, however, the mean scores using the two scaling criteria were different significantly. The norms of the Sichuan population were more similar to the Hong Kong norms than the US norms. The scores of the SF-36 were different between different age and sex groups and between urban and rural populations. CONCLUSION: We have developed the norms of SF-36 by age/sex groups in urban and rural population of Sichuan, China. PMID- 12733353 TI - [The abnormal changes of apolipoprotein(s) in patients with type 2 diabetes mellitus]. AB - OBJECTIVE: To investigate the changes of the apolipoproteins(apoA I, A II, B100, C II, C III, E) in patients with type 2 diabetes mellitus. METHODS: The levels of fasting plasma glucose(FBG), insulin, TG, TC, apoA I, A II, B100, C II, C III, E were all measured in 127 non-diabetic subjects and 143 type 2 diabetic patients (20 associated with coronary heart disease (CHD) and 66 associated with hypertriglyceridemia(HTG), 55 associated with hypercholesterolemia). RESULTS: In male type 2 diabetic patients, the levels of FBG, WHR and apoC II were significantly higher (P < 0.05) and apoA I, A II levels were significantly lower (P < 0.05) than those in male non-diabetic subjects. In female type 2 diabetic patients, the levels of FBG, BMI, WHR, TG significantly elevated while HDL-C, apoA I, A II levels significantly decreased as compared with those in female non diabetic subjects. In type 2 diabetic group, the levels of WHR, FBG and TG in HTG patients were elevated significantly as compared with those without HTG, and the levels of HDL-C, apoA I and apoA II were decreased; the levels of WHR, TG, TC, apoB100, C II, C III, E in patients with HTC were significantly higher than those whose cholesterol levels were normal. In patients with CHD, the levels of fasting insulin, apoB100, apoC II and apoE were significantly higher than those in patients without CHD, and the levels of HDL-C and apoA II were decreased significantly. CONCLUSION: Abnormal changes of apo(s) in type 2 diabetes mellitus may be a cause of type 2 diabetes associated with HTG and CHD. PMID- 12733354 TI - [Effects of retinoic acid on secretion of apolipoproteins A I, A II, B100, C III and E by cultured HepG2 cells]. AB - OBJECTIVE: This study inquired into the role of retinoic acid in lipoprotein metabolism. METHODS: We observed the effect of retinoic acid on the secretion of apolipoproteins A I, A II, C III, B100 and E by cultured HepG2 cells. The apolipoprotiens contents in culture media were measured by radioimmunodiffusion assay (RID) kits developed by our research laboratory. 20-fold lyophilizely condensed culture media were used for the assays. RESULTS: Retinoic acid increased the secretion of apoA I, B100, C III and A II, and it inhibited the secretion of apoE. The effect of retinoic acid was strengthened in a dose dependent manner. When the concentration of retinoic acid in cultured media was 2 x 10(-4) mol/l, the secretion of apoA I, A II, B100 and C III increased by 14.3% (P < 0.01), 23.8% (P < 0.05), 16.1% (P < 0.01) and 47.6% (P < 0.01) respectively, and the secretion of apoE decreased by 37.2% (P < 0.01). CONCLUSION: These findings indicate that retinoic acid does not have a general effect on apolipoprotein secretion in HepG2 cells. PMID- 12733355 TI - [A kinetic study on the relationship between of IL-5, IL-10 and eosinophil apoptosis in asthmatic airway inflammation]. AB - OBJECTIVE: To observe time-course profile of eosinophil apoptosis, in murine asthma models in airway explicate its implication for asthmatic inflammation remission, and investigate the role of IL-5 and IL-10 in modulating cell apoptosis. METHODS: An animal model of asthma was established by OVA sensitizing challenging BALB/c mice. Histologic study of lung tissue was made with the use of electronmicroscope. At seven points (0 h, 8 h, 24 h, 48 h, 96 h, Day 7 and Day 14) in the time course after challenge, bronchoalveolar lavage (BAL) was performed obtain BAL cells and recover fluid. Levels of IL-5 and IL-10 were detected by ELISA. Cell apoptosis was assayed by PI stain on a flow cytometer. RESULTS: After challenge, apoptosis of eosinophils, lymphocytes, endothelial cells could be seen under the electronmicroscope. IL-5 in BALF elevated significantly whereas no significant difference was found between the levels of IL-10 in BALF before and after challenge. Eosinophil apoptosis was identified and a two-phase elevation of apoptosis percentages was observed, although no significant difference was found between average apoptosis in asthmatic group and that in control. Data analyses revealed a negative correlation between apoptosis percentages and the levels of IL-5, and a closer correlation between the percentages and IL-5/IL-10 ratio. CONCLUSION: IL-5 and IL-10 could up- and down regulate asthmatic airway inflammation, respectively. EOS underwent apoptosis in situ and this was modulated by IL-5 and IL-10 interaction. PMID- 12733356 TI - [99mTc-Q3 uptake property at varying coronary flow rates in isolated rabbit heart: comparison between 99mTc-Q3 and 201Tl]. AB - OBJECTIVE: This experiment was designed to explore myocardial 99mTc-Q3 uptake property. METHODS: 20 New Zealand White rabbit heart perfusion models were prepared and the isolated rabbit hearts were perfused by double-radionuclide method at varying coronary flow rates from 0.52 to 3.75 ml/g wet wt/min using developed perfusion liquid with autoblood in comparison with 201Tl in the absence of tracers recirculation. RESULTS: Mean 99mTc-Q3 peak instantaneous uptake was lower(0.65 +/- 0.077) and more affected by low rates (P < 0.05) than 201Tl peak instantaneous uptake (0.82 +/- 0.07, P < 0.01). CONCLUSION: These data were interpreted to indicate: 1. 99mTc-Q3 is not as well extracted as 201Tl; 2. varying the coronary flow rate has significant effects on the uptake of 99mTc-Q3 and 201Tl; 3. similarly to 201Tl, the coronary blood flow conditions may be overestimated by the myocardial radioactive uptake acquired with low flow perfusion, but the coronary blood flow conditions may be underestimated by the myocardial radioactive uptake acquired with high flow perfusion. Further studies on retention and clearance of 99mTc-Q3 need to be conducted. PMID- 12733357 TI - [Effect of high glucose condition on the secretion of TGF beta by cultured macrovascular endothelial cells of the diabetic rat]. AB - OBJECTIVE: This study was designed to examine the secretion of TGF-beta by cultured rat thoracic aortic endothelial cells of the diabetic rats under high gluocose condition. METHODS: First we created diabetic rat models, then the thoracic aortic endothelial cells of the diabetic rats were isolated and cultured in 10% FBS-RPMI 1640 media with various high glucose concentrations (12.5 mmol/L, 25 mmol/L or 50 mmol/L D-glucose), high glucose and TNF-alpha or high glucose and insulin for 48 hours, and the supernatant obtained from the cultured aortic endothelial cells was collected. The level of TGF-beta 1 in the supernatant was measured using a standard bioassay, which utilizes the TGF-beta 1 sensitive Mv1Lu (CLL-64) Mink lung epithelial cell line, and the production of endothelin (ET) in the supernatant was also assessed by RIA (radio immol/Lunoassay). RESULTS: Both total TGF-beta 1 (406 +/- 29 vs 189 +/- 23 pg/ml, P < 0.05) and active TGF-beta 1 (161 +/- 24 vs 39.5 +/- 8 pg/ml) secreted by the cultured aortic endothelial cells under high glucose condition (25 mmol/L) were significantly increased as compared with control. TNF-alpha could enhance the effect of high glucose on the secretion of TGF-beta 1, whereas insulin could inhibit it. With various high glucose concentrations, the levels of ET were increased. Also TNF-alpha enhanced it and insulin inhibited it. CONCLUSION: TGF-beta 1 may be involved in the development of diabetic macrovascular complication. PMID- 12733358 TI - [Effect of human umbilical vein endothelial cells on granulopoiesis, erythropoiesis and megakaryopoiesis in mice]. AB - OBJECTIVE: To investigate the effects of endothelial cells on the proliferations of granulocytic, erythrocytic and megakaryocytic progenitor cells. METHODS: Human umbilical vein endothelial cells were collected within 12 hours after cesarean section and were cultured according to Jaffe's method. The human endothelial culture supernatant (HECS) harvested from primary culture medium was centrifuged. Bone marrow mononuclear cells of mice were prepared and then cultured in semi solid agar or methylcellulose with the addition of HECS. RESULTS: Colony-forming yields were increased in the culture systems with HECS. But in the systems with no HECS, exogenous GM-CSF, EPO and IL-3 added to the hematopoietic progenitor cultures were necessary to induce colony-forming. Synergistic increase in progenitor cell expansion was observed when both HECS and exogenous cytokines were added to hematopoietic progenitor cultures. CONCLUSION: 1. Endothelial cells can support the proliferation and differentiation of hematopoietic stem and progenitor cells. 2. HECS as a source of standard hematopoietic growth factors can be used in the expansion of stem cells in vitro and in the therapy of hemopathy. 3. Interactions between endothelial cells and hematopoietic stem cell or progenitors may play an important role in the regulation of hematopoiesis in vivo. 4. Endothelial cells may produce other soluble growth factors potentiating the action of a set of cytokines such as GM-CSF and IL-6. PMID- 12733360 TI - [Relationship between apoptosis and expression of Fas, bcl-2 and Ki-67 in condyloma acuminatum]. AB - OBJECTIVE: This study inquired into the significance and relationship of apoptosis to the regulation and proliferation of condyloma acuminatum(CA). METHODS: Apoptotic cells were detected in situ by terminal deoxynucleotidyl transferase-mediated-dUTP nick end labeling (TUNEL). The detection of Fas, bcl-2 and Ki-67 expression was performed using immunohistochemical staining in 28 specimens of CA. RESULTS: Apoptotic cells were found in 24 cases (85.7%) in keratinocytes. All of the in 28 cases(100%) had Fas and Ki-67 positive expressions, but only 8 cases (28.6%) had bcl-2 mild or moderate expression. The disease severity (serious, moderate and mild) groups did not significantly differ in apoptotic index (AI), Fas, bcl-2 and Ki-67 positive expression (P > 0.05). There was positive correlation between AI and Fas(r = 0.866, P = 0.000). bcl-2 and Ki-67 were negatively correlated with AI (r = -0.416, P = 0.018; r = -0.475, P = 0.006). CONCLUSION: It is suggested that apoptosis in keratinocytes may play an important role in inhibiting the proliferation of virus, that Fas may be an upregulative factor and bcl-2 may be an inhibition factor to apoptosis, and that excessive Ki-67 expression may be associated with the acceleration of cell cycle. PMID- 12733359 TI - [Histopathological observation of acute facial nerve impairment in rabbits]. AB - OBJECTIVE: To investigate histopathological changes of acute facial nerve impairment. METHODS: Ligation of facial nerve outside stylomastoid foramen with 3 0 silk, observation of histopathological changes of facial nerve on the 1st day, 3rd day and 5th day after ligation. RESULTS: The facial nerve showed all kinds and various degrees of histopathological changes, from swelling to degeneration, in myeline and axon respectively. CONCLUSION: All kinds and various degrees of pathological changes have been observed in myeline and axon after the ligation of facial nerve in rabbits. This study demonstrates that such injuries to the facial nerve become increasingly serious as the ligation lasts day after day. PMID- 12733361 TI - [Effect of increased and decreased bite force on morphology of periodontal tissues]. AB - OBJECTIVE: The aim of this study was to observe the effect of bite force on morphology of periodontal tissues. METHODS: Animal models were induced by extracting the left maxillary molars; the left mandibular molars served as decreased bite force models; the right mandibular molars served as increased bite force models. The dynamic changes of widths of periodontal ligament and cementum were measured. RESULTS: In decreased bite force group, there were significant morphologic changes of periodontal tissues after 1 week. In increased bite force group, there was no significant change. Ligament widths were significantly different after 2, 3 weeks respectively in those two groups, compared with that in control group (P < 0.05). There were significant differences between proximal alveolar wall and distal alveolar wall in width (P < 0.05). CONCLUSION: The histological morphology is closely related to the mechanical condition. Physiological bite force is necessary for periodontal ligament to maintain its physiological structure. PMID- 12733362 TI - [The mechanism of protective effects of enalapril on experimental diabetic nephropathy of rats]. AB - OBJECTIVE: To elucidate the mechanisms of renoprotective effects of ACEI on diabetic animals and patients. METHODS: STZ-induced diabetic rats were treated with Enalapril and the levels of urine protein, TGF-beta and AGEs in renal tissue were measured and compared. RESULTS: The results of TGF-beta quantitation and AGE fluorescence of Enalapril treatment group rats were the same as those of diabetic control rats. CONCLUSION: Suppression of TGF-beta overexpression or AGEs accumulation was not implicated in the mechanisms of renoprotective effects of ACEI. PMID- 12733363 TI - [Lymphocyte subpopulation, interleukin-2 and high affinity interleukin-2 receptor expression in primary nephrotic syndrome]. AB - OBJECTIVE: To understand the cellular immune response during relapse and remission stage in primary nephrotic syndrome (PNS). METHODS: We applied the radioligand binding assay (RBA), the bioactivity measurement of IL-2, and monoclonal antibody sensitized red blood cell method to evaluate the expression of IL-2R, the production of IL-2 and T lymphocyte subsets of PBMC from 22 patients suffering from PNS, 17 patients with PNS in remission, and 25 normal subjects matched age and sex. RESULTS: CD3, CD4, CD8, IL-2, IL-2R in relapse were significantly lower than those in control (P < 0.05); all above except CD8 in remission patients were significantly higher than those in relapse, but lower than those in control (P < 0.05). CONCLUSION: This study suggested that the cell mediated immunity (CMI) during acute nephrotic phase decreased; the deficiency of CMI of PNS in remission improved a lot when compared with the acute phase, but still not recovered completely; the deficiency of CMI with PNS in remission might be one of the causes of easily recurring of PNS. PMID- 12733364 TI - [Study on hypermethylation of the calcitonin gene in malignant hematological disorders]. AB - OBJECTIVE: To investigate the clinical significance of hypermethylation of the calcitonin (CT) gene in malignant hematological disorders (MHD). METHODS: Polymerase chain reaction (PCR) in combination with digestion of DNA with Hpa II was used to examine the methylation patterns of the CT gene in 73 patients with MHD, 6 healthy people and 24 patients with nonmalignant hematological disorders (NON-MHD). RESULTS: Hypermethylation of the CT gene occurred in 12 of 14 (85.7%) patients with acute lymphoblastic leukemia (ALL), 9 of 15 (60%) acute nonlymphocytic leukemia (ANLL), 8 of 10 (80%) chronic myelogenous leukemia (CML), 5 of 15 (33.3%) malignant lymphoma (ML), 2 of 5 patients with myelodysplastic syndrome (MDS), 1 of 2 malignant histiocytosis (MH), 1 of 3 chronic lymphocytic leukemia (CLL) and 1 of 9 multiple myeloma (MM). However, it did not occur in normal controls and NON-MHD. CONCLUSION: Hypermethylation of CT gene may serve as an important molecular maker for tumorous cell clone and may be useful in diagnosing MHD, monitoring minimal residual disease and predicting prognosis. PMID- 12733365 TI - [Level of nerve growth factor protein in cerebellar cortex of rat pups in intrauterine growth retardation model by clamping the uterine vasculature of pregnant rat]. AB - OBJECTIVE: To understand the level of nerve growth factor (NGF) in the brain of rat pups in intrauterine growth retardation (IUGR), explore the mechanism of brain injury. METHODS: Animal models for IUGR pups were established by clamping the uterine vasculature of pregnant rats with 30 minutes in this study. NGF proteins were measured by immunohistochemistry in the cerebellar cortex of rat pups at birth (21 days of gestation), and on the sixth day after birth. RESULTS: 1. The body weight of the 21-day fetuses in IUGR group decreased 16.1% as compared to that in control group (P < 0.0001); 2. The brain weight of the 21-day fetuses in IUGR group decreased to that in control group (P < 0.05) development of their brain retardation during the intrauterine in IUGR fetus period in this model; 3. On the sixth day after birth, there was no statistical difference in body weight between the IUGR and the control groups. But the brain weight difference between these two groups was still there; 4. At birth and on the sixth day after birth, there were statistical differences in NGF proteins in cerebellum between the IUGR and the control groups, which got along with the decrease of their brain weights. CONCLUSION: A successful animal model of IUGR was established by clamping the uterine vasculature of pregnant rats with 30 minutes, and the development of their brain were retarded in IUGR fetus period in this model. Brain retardation in IUGR pups may relate with the decreased level of NGF in brain. This study discovered that quick "catch-up" growth in physical development and slow "catch-up" growth in brain development in IUGR pups. PMID- 12733366 TI - [Impact of WPY on pancreatic microcirculation of acute pancreatitis in mice]. AB - OBJECTIVE: To explore the pathogenesis of acute pancreatitis (AP) and evaluate the effect of a Chinese herb WPY on the course of AP. METHODS: Intravital fluorescence microscopy was used. The pancreatic microvascular diameter, RBC velocities and functional capillary density (FCD) were estimated following intracutaneous injection of caerulein alone or with gastropipe WPY. RESULTS: Caerulein mediated a significant decrease in functional capillary density (FCD), RBC velocities and diameter of interlobular arteriole (P < 0.01). Compared with AP group, WPY was effective in restoring functional capillary density, interlobular arteriole diameter and RBC velocity. CONCLUSION: Impairment of pancreatic microcirculation in the early phase of acute pancreatitis may play a key role in the progression of this disease. Possible contributory mechanisms include reduced blood flow and functional capillary density, interlobular arteriole spasm, and leukocyte-endothelial cell interaction. WPY has a beneficial effect on the course of acute pancreatitis. Possible causes include attenuating microcirculatory failure. PMID- 12733367 TI - [Effects of calcium channel antagonist on gastrin-induced proliferation of HT29 colon carcinoma cells]. AB - OBJECTIVE: This study aimed to assess the effects of nifedipine on gastrin induced proliferation of HT29 colon carcinoma cells and inquire into the possible mechanisms. METHODS: Flow cytometry was used to monitor the cytoplasmic free calcium; MTT colorimetry was used to determine the proliferation of HT29 cells. RESULTS: The results showed that 2.5 x 10(-6) mol/L pentagastrin (PG) induced a quick rise of intracellular free calcium ([Ca2+]i) (P < 0.01). 10(-5) mol/L nifedipine can significantly inhibited the rise of [Ca2+]i induced by pentagastrin (P < 0.01), in parallel, at growth assay we demonstrated that 10(-5) 10(-6) mol/L nifedipine could obviously block the increase in cell number elicited by 2.5 x 10(-6) mol/L PG. CONCLUSION: These data indicate that nifedipine can stop the influx of Ca2+ and hence inhibit the pentagastrin-induced proliferation. PMID- 12733368 TI - [Comparison between human sperm zona-free hamster ovum penetration assay and in vitro fertilization]. AB - OBJECTIVE: The research question was whether the result of human sperm zona-free hamsterovum penetration assay (SPA) could serve as an indicator for in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). METHODS: The semen samples of 22 male partners in couples with infertile marriage were assessed for their fertilizing capacity by SPA. The fertilization rate of SPA was compared with that of IVF. RESULTS: The fertilization rates of SPA and IVF were 38.6% +/- 23.5% and 53.5% +/- 28.4% respectively, and a significant correlation between SPA and IVF was observed (rs = 0.7045). CONCLUSION: The result of SPA may be used as a reference indicator for assisting fertilization, IVF or ICSI. PMID- 12733369 TI - [Changes of tangent time constant in the transformation of F-V curve configuration in COPD and cor pulmonale patients and clinical implications]. AB - OBJECTIVE: To explore the changes of tangent time constant (tau t) in the transformation of F-V curve configuration in COPD and cor pulmonale patients and clinical implications. METHODS: We determined the F-V curves in 90 COPD patients (43 with chronic bronchitis, 47 with emphysema) and 31 complicated with cor pulmonale; all patients were in the ameliorated period. The tau t at high, mid and low lung volumes, tau t75, tau t50, tau t25, were measured on each F-V curve. RESULTS: 1. Normal ranges: F-V curve configuration with plateau, linear, convex and concave types; |tau t75| > ls or < ls; tau t25 < ls; tau t relationship with "adolescent fashion" or "old fashion". 2. Light and moderate abnormality: With convex type; |tau t75| < ls; tau t25 approximately = ls or > ls; "trumpet fashion" changes. 3. Serious abnormality: With hyperbolic type; |tau t75| < ls; tau t25 = 4s-10s; "tower fashion" changes. CONCLUSION: Hyperbolic type, extreme prolongation of tau t25 and "tower fashion" changes can be assigned as 3 important indices to assess the severity of disease. The convenience and correctness of the method may be available for clinical application. PMID- 12733370 TI - [Smoking and alcohol consumption as risk factors of pulmonary tuberculosis in Chengdu: a matched case-control study]. AB - OBJECTIVE: The aim of this study was to assess the relationship between personal behaviors (smoking and alcohol consumption) and contracting pulmonary tuberculosis. METHODS: 346 persons (173 cases and 173 controls) were selected from 12 communities of Chengdu area, all the cases were active TB patients (by WHO criteria) from March 1996 to March 1997. Controls were matched for age, sex and living district. Subjects were interviewed face to face by trained interviewers using questionnaires. RESULTS: The results of univariate analysis showed that active smoking (OR = 2.12, P = 0.006), passive smoking (OR = 1.55, P = 0.04), type of cigarettes (OR = 1.31, P = 0.005) and alcohol consumption (OR = 1.81, P = 0.008) were significantly associated with TB. Yet, multivariate logistic regression analysis did not find smoking or alcohol consumption being in independent association with TB, but it showed that persons who were smokers with the addition of alcohol consumption had a higher risk to contract TB (OR = 7.729, 95% C.I. = 1.5215-39.2634). Significant association was noted in the dose response analysis (OR = 1.73, 95% C.I. = 1.300-2.3028). CONCLUSION: These data indicate that smoking alone or sole alcohol consumption bears no relationship with TB, but smoking plus alcohol abuse is probably a risk factor for pulmonary tuberculosis in Chengdu, and in this connection, a proposal of prospective study to further demonstrate this risk factor is warranted. PMID- 12733371 TI - [Analysis on the sex ratios at birth for deliveries in hospitals in China from 1987 to 1992]. AB - OBJECTIVE: To estimate the sex ratios at birth (SRB) for livebirths in hospitals in China. METHODS: Data were collected by the birth defects monitoring network. The livebirths with gestation of 28 weeks or over delivered in hospitals of 30 provinces, municipalities and autonomous regions all over the country from 1987 to 1992 were analyzed and used for calculating the SRB. RESULTS: The high male bias in SRB was observed in the data from most areas, and there was an upward trend of SRB, especially in rural areas. The SRB for livebirths in the hospitals of urban areas was normal from 1987 to 1989, and rose to 108.9 in 1992. The SRB for rural areas was high and was 109.9 in 1987, and rose to 111.8 in 1992. The SRB for livebirths given by the mothers under 20 years of age and coming from lower educational background was higher than that for others. The SRB for higher parities was even higher. CONCLUSION: The SRB in China is high; this is possibly related to sex-preference. Under-reporting of female neonates and "missing females" are not the main causes of the rising SRB. PMID- 12733372 TI - [A multicenter randomized controlled clinical trial on lipids regulating effects of domestic simvastatin]. AB - OBJECTIVE: This clinical trial was designed to assess the lipids regulating effects of domestic simvastatin (DS, produced by Chengdu Huayu Pharmaceutical Co.) in patients with hyperlipidemia. METHODS: 160 hyperlipidemic patients were randomly divided into 3 groups (A, B and C). Groups A and B were subjected to single-blind trial; group C was for open trial. Group A took DS 10 mg q.n., group B Zocor 10 mg q.n. and group C DS 10 mg q.n. respectively for 8 weeks. All the patients were followed up at the 4th week and 8th week. 155 patients finished the trial with 59 cases in group A, 47 cases in group B and 39 cases in group C. RESULTS: At the 4th week, serum total cholesterol (TC) in group A, B and C decreased by 16.88%, 19.23% and 14.10%; serum triglycerides (TG) decreased by 19.27%, 15.66% and 17.96%; HDL-C increased by 7.69%, 7.46% and 6.69%; and LDL-C decreased by 23.02%, 27.84% and 24.43%, respectively; there was no significant difference among the three groups (P > 0.05). At the 8th week, serum TC in groups A, B and C decreased by 25.03%, 26.53% and 25.22%. TG decreased by 23.85%, 24.74% and 24.75%; HDL-C increased by 9.23%, 8.95% and 8.39%; and LDL-C decreased by 33.72%, 35.50% and 30.99%, respectively; still, no significant difference among the three groups was observed (P > 0.05). The incidence rates of side effects in the three groups were similar. The clinical effects were more significant at the 8th week than at the 4th week for Zocor and DS. CONCLUSION: These data suggest that DS is as effective and safe as Zocor in clinical use for lipids regulating serum. PMID- 12733373 TI - [A multi-centered clinical study an domestic intravenous Turbutalin sulfate in the treatment of asthma and chronic asthmatic bronchitis]. AB - OBJECTIVE: This multi-centered clinical trial was designed to assess the efficacy and safety of domestic intravenous Turbutalin Sulfate in the treatment of asthma and chronic asthmatic bronchitis. METHODS: Bricanyl was used as control. A total of 120 patients were included in this randomized controlled study. The two medications, both at a dose of 0.25 mg intravenous drip, were given three times a day for 3-5 days. RESULTS: The two drugs' clinical excellent rates were 66.7% and 73.3%; overall efficacy rates were 98.3% and 100.0%, respectively. Their pulmonary functional excellent rates were 78.4% and 76.6%; overall effect rates were 88.3% and 90.0%, respectively. Composite curative evaluation showed that the two drug's composite excellent rates were 61.6% both; their overall efficacy rates were 88.3% and 90.0%, respectively. All of these showed no significant difference (P > 0.05). The incidence rates of adverse side effects of the two groups were 13.5% and 15.0%, respectively; most of them were mild and tolerable. CONCLUSION: Domestic intravenous Terbutalin Sulfate is effective and safe in treating asthma and chronic asthmatic bronchitis. PMID- 12733375 TI - [Expression of C-myc and N-myc protein in adulthood and childhood medulloblastomas and prognostic analysis]. AB - OBJECTIVE: To detect the expression of C-myc and N-myc in medulloblastomas, the difference between adults and children, and the relation between expression and prognosis. METHODS: ABC immunostaining was used to detect C-myc and N-myc protein expression in 21 cases of adulthood medulloblastomas and 14 childhood cases. Follow-up materials were obtained to perform survival curve analysis. RESULTS: Of these 35 cases, 5 (4 children, 1 adult) showed C-myc positive expression. No N myc expression was found. There was no relationship between expression and survival. CONCLUSION: C-myc may play a role in the tumorgenesis of medulloblastoma, especially in children, but it is of no prognostic significance. N-myc antigen may be lost during the fixation of specimens. PMID- 12733374 TI - [Inflammatory mediator in patients with obstructive jaundice and its effect on renal function]. AB - OBJECTIVE: To investigate the effect of inflammatory transmitter on renal function in obstructive jaundice. METHODS: 15 patients with obstructive jaundice (OJ) as the experimental group and 15 non-joundiced patients as the control group were involved in this study. The plasma ET, LPS, TNF-alpha, IL-6, IL-8 and urine REP, TFR, ALB were measured in 15 patients with OJ before and 1, 4, 7 days after surgery and in 15 patients without jaundice as controls. RESULTS: The levels of plasma ET, LPS, TNF-alpha, IL-6, IL-8 and urine RBP, TFR, ALB increased significantly at multiple time points as compared with those in the non-OJ patients (P < 0.05), and the changes of plasma ET, LPS, TNF-alpha, IL-6, IL-8 are the same as those of urine RBP, TFR, ALB. CONCLUSION: These data suggest that there is interaction between ET, LPS, TNF-alpha, IL-6, IL-8 and the damage to renal function with OJ after surgery. PMID- 12733376 TI - [The value of three-dimensional helical CT imaging in the diagnosis of maxillofacial fractures]. AB - OBJECTIVE: To assess the diagnostic value of three-dimensional (3D) helical CT imaging in the cases of maxillofacial fractures. METHODS: 25 trauma patients were examined with thin-slice CT scan and 3D reconstruction. RESULTS: Three dimensional images reconstructed from helical CT clearly and stereoscopically demonstrated the location, shape and extension of maxillofacial fractures. CONCLUSION: 3D helical CT imaging can provide valuable information in demonstrating space relationships of maxillofacial fractures. PMID- 12733377 TI - [Local recurrence of nasopharyngeal carcinoma with normal mucous membrane after radiation therapy: MR imaging findings]. AB - OBJECTIVE: The purpose of this work was to investigate and analyze the MR imaging features of postradiation local recurrence of nasopharyngeal carcinoma (NPC) with normal mucous membrane. METHODS: 24 such cases were studied. The diagnosis of NPC recurrence was pathologically confirmed in 15 patients, and was corroborated by repeated radiation therapy and serial follow-ups in 9 patients. MR imaging routine procedures and Gd-DTPA enhancement were performed for all patients. RESULTS: The MR imaging findings were as follows: 1. The recurrent tumor mass was located in the prestyloid space in 8 patients, with skull base erosion and intracranial infiltration in 7 patients; 2. Retrostyloid space recurrence in 2 patients; 3. Direct bony erosion of the skull base in 7 patients; 4. Recurrence in nasal cavity and ethmoid sinuses in 3 patients; 5. Submucous cystic recurrence in 2 patients; 6. Mixed patterns of recurrence in 2 patients. Except 2 cases of cystic tumor recurrence, all cases exhibited slight hypo- or iso-intensity on T1 weighted images, slight hyper-intensity on T2-weighted images, and moderate to marked enhancement after Gd-DTPA administration. CONCLUSION: There are several special patterns of local NPC tumor recurrence after radiation therapy in the presence of normal nasopharyngeal mucosa. MR imaging is the method of choice to depict the location and extent of NPC tumor recurrence. PMID- 12733378 TI - [Study on bolus cyclosphamide treatment for 64 cases of lupus nephritis]. AB - OBJECTIVE: To report on the dosing, efficacy and side-effects of bolus cyclosphamide treatment for lupus nephritis (LN). METHODS: 64 consecutive cases of LN with 10 or more erythrocytes per high-power field, proteinuria (> 1 g of protein per day) and serum creatinine increased (> 133 mumol/L) were treated by bolus therapy with cyclophosphamide (CTX) given monthly for 6 months and then quarterly for 18 months. RESULTS: 49 patients had renal remission (defined as < 10 erythrocytes per high-power field, absence of cellular casts, excretion of < 1 g of protein per day and normal serum creatinine). The mean of doses was 1.1 g for each time (0.6-1.6 g), the mean of times of bolus CTX needed was 3.6 (1-8 times). The adverse events were amenorrhea (in 41% female patients), herpes zoster (in 13% patients) and hemorrhagic cystitis (in 1 patient). CONCLUSION: The results indicate that monthly bolus CTX therapy is effective and safe for patients with LN. Its adverse effect is relatively not a serious problem. PMID- 12733379 TI - [Investigation on the value of applying interventional radiologic treatment in the management of advanced gynecological malignancies]. AB - OBJECTIVE: To investigate the value of applying interventional radiologic treatment (IRT) in the management of advanced gynecological malignancy. METHODS: Twenty-four historically confirmed advanced gynecological malignancy patients from September 1997 to June 1998 were included. After receiving internal iliac arterial infusion chemotherapy with Seldinger, the patients' tumor tissues were pathologically studied at different times. The tumor size was measured by ultrasonography two weeks after IRT. RESULTS: Before internal iliac arterial infusion chemotherapy, internal iliac arteriography showed that uterus artery was thickened and tortuous, the new vessels appeared and the tumors were stained. In some cases, the image of blood vessel-lake and arteriovenous shunt were seen in tumor tissues. Twenty-four hours after internal iliac arterial infusion chemotherapy, focal necrosis was observed in tumor cells, seventy-two hours after, the necrosis became more thorough. In regard to the tumor size, two patients were CR, nineteen patients were PR, and two patients were NC. Only one patient failed to respond to IRT. Chemotherapy side effects were slight. Patients' WBC returned normal within two weeks after chemotherapy. No complication happened. CONCLUSION: 1. Internal iliac arteriography has practical significance in assessing the tumor size and evaluating the therapeutic effects; 2. the internal iliac arterial infusion chemotherapy can reduce the tumor size, which prepares the patient in a better condition for operation. The most appropriate time for operation is two weeks after internal iliac arterial infusion chemotherapy. Even for the patient who is not suitable for operation, the reduction of the tumor size still can enhance the therapeutic effects of radiotherapy. 3. IRT is easy to perform, effective and with few slight side effects and complication. It is an effective way to save the patient's life. PMID- 12733380 TI - [Study on the chemosensitivity test of human gastric cancer using the MTT assay]. AB - OBJECTIVE: This study was conducted to investigate the chemosensitivity of gastric carcinoma. METHODS: 112 cases of fresh human gastric cancer samples were obtained for drug sensitivity testing using MTT. RESULTS: The sensitivity sequence of antitumor agents was 5-Fu, DDP, MMC, etc. On the other hand, the sensitivity was relative to pathology and individual differences. CONCLUSION: The MTT assay is an accurate system for determination of the chemosensitivity of tumor cells. PMID- 12733381 TI - [Clinical application and evaluation of polymerase chain reaction-microwell plate hybridization assay in the detection of Mycobacterium tuberculosis]. AB - OBJECTIVE: To evaluate the clinical application and feasibility of polymerase chain reaction-microwell plate hybridization assay in the detection of Mycobacterium tuberculosis. METHODS: 1130 specimens with strong suspicion for mycobacterium tuberculosis were collected from the hospitals and were detected by fast bacilli stain, culture, PCR-electrophoresis and PCR-microwell plate hybridization respectively. The laboratory results were analyzed in combination with the symptoms and signs of patients and the observations on treatment. Also detected were 100 samples from the clinically evidenced non-tuberculosis patients. RESULTS: In the 100 samples collected from the patients without tuberculosis, the PCR-hybridization method and culture method did not detect Mycobacterium tuberculosis, but the fast bacilli stain method and PCR electrophoresis method brought out one and two false-positive results respectively. The results of testing the 1030 clinical samples which probably contained Mycobacterium tuberculosis demonstrated that the PCR-hybridization method had the highest positive rate (481/1030) among the four methods, and the positive rates of the other three methods were PCR-electrophoresis (406/1030), culture (365/1030) and fast bacilli stain (256/1030) in proper order. The chi square test showed that there were significant difference between the PCR hybridization method and the other three methods respectively (P < 0.0083 or P < 0.0017). CONCLUSION: PCR-Hybridization method is specific, sensitive, accurate and fast in detecting mycobacterium tuberculosis; it is worthy to be used clinically. PMID- 12733383 TI - [Determination of warfarin in plasma by HPLC and an investigation of monitoring patients after cardiac valve replacement]. AB - OBJECTIVE: A simple HPLC method was established for the determination of warfarin in plasma to investigate the relationship between warfarin concentration and anticoagulant effect. METHODS: The mixture of dichloromethane and hexane (1:9) was used as extracting solvent for the plasma samples. The chromatographic separation was on C18 column with a mobile phase consisting of methanol and 50 mmol/L ammonium acetate buffer (pH2.5, 70:30). RESULTS: The calibration curve was linear within 50-2000 ng/ml. The extraction recoveries of warfarin were 78.0% 81.6%. The recoveries of methodology were 103.1%-106.5%. Inter-day and intra-day RSD were 2.33%-5.46% and 5.29%-7.73%, respectively. This method was used for determining warfarin in 70 patients after cardiac valve replacement. The results showed that 44 cases had their INR within the safety range (1.31-2.35) recommended to Chinese, and of them 37(84.1%) cases had a warfarin level at 616.2 +/- 154.8 ng/ml. CONCLUSION: This method is useful in monitoring warfarin concentration during anticoagulant therapy. PMID- 12733384 TI - [Is it of value to read medical articles?]. PMID- 12733382 TI - [Improved APAAP immunocytochemical staining-trace cytotoxicity test plate method for detection of cell surface antigens]. AB - OBJECTIVE: To improve the conventional slide method of alkaline phosphatase-anti alkaline phosphatase (APAAP) immunocytochemical staining for detection of cell surface antigens. METHODS: Trace cytotoxicity test plates were utilized in the detection of CD3, CD4, CD8, CD11a, CD11b, CD18 and CD54 on lymphocytes by APAAP immunocytochemical staining. The percentages of positive cells were counted. Meanwhile, slides were used in the same objects and procedure. RESULTS: The results of trace test plate method were similar to those of slide method (P > 0.05, respectively). The reproducibility was very excellent. The plate method decreased the volume of blood from 10 ml to 2 ml to detect 7 norms, and each reagent from 50 microliters to 5 microliters. CONCLUSION: Trace test plate method may not only attain the result similar to that of the slide method, but also save blood and reagents; it is easy to do, so it is more suitable for clinical examination and scientific research in hospitals or institutions at the basic level. PMID- 12733385 TI - [Cancer study and the Academy of Medicine of the 20th century]. PMID- 12733386 TI - [Terminal care of patients by the general practitioner and the specialist. Results of a French survey "Attitudes and practices in palliative treatment- 2002"]. AB - OBJECTIVE: Assess the attitude of the practitioners with regard to the management of patients at the end of their lives, depending on the physician/patient communication and treatment of pain. Method Statistical analysis of the data collected among 1,000 practitioners recruited for a survey on knowledge, attitudes and practices regarding palliative care in 2002. RESULTS: The practitioners who felt "ill at ease" in the management of patients at the end of their life often had a poor opinion regarding morphine and did not systematically their patients of the therapeutic aims. CONCLUSION: Greater awareness by the practitioners of palliative care would help to improve the quality of care supplied to terminally ill patients. PMID- 12733389 TI - [Pseudotumor form of bronchial tuberculosis]. PMID- 12733387 TI - [Control of asthma by treatment with inhaled corticosteroids and prolonged action beta 2-agonists in free or fixed combination. Results of the ALISE study]. AB - CONTEXT: The basics of asthma treatment are well defined in the guidelines, however the criteria for the choice between fixed and free combinations of inhaled corticosteroids and long acting beta 2-agonist have not been clearly established. METHOD: A modified Juniper questionnaire (using peak expiratory flow rather than maximal expiratory volume per second) was used by 421 general practitioners to assess asthma control in 861 adult patients (aged 47 +/- 17 years) having received inhaled corticosteroids and long-acting beta 2-agonist in a fixed (n = 413) or free (n = 448) combination for at least 6 weeks. RESULTS: Treatment groups were comparable for demographics, asthma history and severity as assessed by the physician. Despite better compliance to treatment in the fixed combination group and a mean daily dose of corticosteroids around two-fold greater compared with free combination group (913 +/- 450 micrograms/d vs 401 +/- 178 micrograms/d), the mean Juniper score was lower in the free combination group (1.60 +/- 0.94 vs 1.73 +/- 0.96) with a greater percentage of patients controlled (Juniper score < 2): 67.6% vs 60.8%. Other treatments for asthma were significantly less frequent with the free combination (14.2% vs 22.3%), as were local side effects and notably voice changes (3.3% vs 9.5%. COMMENTS: The apparently greater efficacy/safety ratio of the free combination was probably due to the greater severity of asthma in the patients treated with a fixed combination. This highlights the interest of free combinations as first-line therapy for patient with mild to moderate asthma. PMID- 12733388 TI - [Febrile pseudotumor lesion of the arm. Atypical presentation of Shulman syndrome]]. AB - INTRODUCTION: The diagnostic approach when confronted with a pseudo-tumoral lesion and fever is difficult since it evokes an infectious, rheumatismal inflammatory or even systemic disease. OBSERVATION: A 39 year-old Vietnamese man was hospitalised for polyarthralgia if the arm and fever (39 degrees C). He was treated with non-steroidal anti-inflammatories, which were only partially effective. Biological examinations revealed a severe inflammatory syndrome and cytolysis three-fold higher than normal. Since the infectious, rheumatismal and immunological explorations were negative and in view of the appearance of a pseudo-tumoral inflammatory lesion of the left arm concomitant to infiltration of the underlying muscle revealed on MRI, a muscle biopsy was performed showing eosinophilic fasciitis. Diagnosis of Shulman's syndrome was made and the affection rapidly improved with corticosteroids. COMMENTS: The presentation of our patient was atypical for several reasons: his Vietnamese origin, the initial fever peak at 39 degrees C, the unilateral involvement, the severe inflammatory syndrome, the initial absence of hypereosinophilia, the visceral involvement and notably myositis are all uncommon in Shulman's syndrome. Hence, it is important to rule out its differential diagnosis from local edematous scleroderma, which does not share the same prognosis. PMID- 12733390 TI - [Association of parathyroid adenoma and malignant blood disease]. PMID- 12733391 TI - [Epidemiologic, clinical, biological and therapeutic aspects of Gaucher disease]. AB - GENERAL CHARACTERISTICS: Gaucher's disease is a genetic disease of autosomal recessive transmission due to a deficit in a lysosomal enzyme: beta glucocerebrosidase. The disease is characterised by deposits of glucosylceramide in the cells of the liver, spleen and bone marrow. Acute or chronic neurological forms (type 2 and 3) account for only 5% of patients suffering from Gaucher's disease and are less frequent than the non-neurological forms (type 1). CLINICAL AND BIOCHEMICAL MANIFESTATIONS: Gaucher's disease is associated with spleno- or hepato-megalia, asthenia, bone complications (Erlenmeyer flask deformity, osteopenia and osteonecrosis), as well as with haematological (thrombopenia, anaemia) or biochemical abnormalities (increase in angiotensin-converting enzyme, ferritin, tartrate-resistant acid phosphatase and chitotriosidase). Central nervous system involvement is only found in the type 2 and 3. Diagnosis relies on measurement of beta-glucocerebrosidase activity in the circulating leukocytes. REGARDING TREATMENT: Treatment with enzyme replacement (imiglucerase: recombinant enzyme preparation) improves the haematological abnormalities, hepatosplenomegalia and quality of life in a matter of a few months. Regression of the bone disorders is usually observed only after 3-4 years of treatment. Recently, gene therapy trials have successfully been started. PMID- 12733392 TI - [From hepatic steatosis due to obesity to cirrhosis, in the absence of alcoholic intoxication]. AB - FROM STEATOSIS TO CIRRHOSIS: Obesity is frequently associated with steatosis. The latter may provoke inflammatory manifestations and steatohepatitis, the clinical biological and histological characteristics of which are identical to lesions of alcoholic origin. A fibrosis may appear after several years, particularly if the obesity is severe, the patient is elderly and suffering from diabetes and that the increase in transaminase exceeds twice the normal value. It is therefore justified to propose a hepatic biopsy in an obese patient when abnormalities in transaminase levels exceed twice the normal values and notably if other fibrosis risk factors exist. THERAPEUTIC ATTEMPTS: Treatment is aimed at correcting the metabolic disorders and the obesity. Other treatments are currently proposed (anti-oxidants, insulin-resistance modulators), but none of them have presently been completely validated. IN THE FUTURE: With the increase in obesity in industrialised countries, the prevalence of steatosis and steatohepatitis will also increase and there is a risk that they will become, within a few years, one of the principle causes of cirrhosis. PMID- 12733393 TI - [Development of antidepressant drugs. Experience and prospects]. AB - THE FIRST ROUTES OF RESEARCH: The first antidepressants were developed after the discovery of the existence during depression of a perturbation in the synaptic transmission of the principle monoamines: noradrenalin, serotonin and dopamine. The pharmacological effect of the various molecules developed is mainly on the metabolisation routes of neurotransmitters, but may also concern the different receptors present on synaptic level. THE AWARENESS OF NEW MEDIATORS: The progress in research on antidepressants has widened the scope of the development of such medicinal products to the domain of endocrinology (hypothalamo-pituitary-adrenal axis, progestogen hormones, thyreotropic axis) and studies on neuropeptides (substance P, neuropeptide Y). The complexity of the physiopathological mechanisms of depression hence appears enhanced. PMID- 12733394 TI - [Pseudo-Pott's disease due to Pseudomonas aeruginosa]. PMID- 12733395 TI - Number of ancestral human species: a molecular perspective. AB - Despite the remarkable developments in molecular biology over the past three decades, anthropological genetics has had only limited impact on systematics in human evolution. Genetics offers the opportunity to objectively test taxonomies based on morphology and may be used to supplement conventional approaches to hominid systematics. Our analyses, examining chromosomes and 46 estimates of genetic distance, indicate there may have been only around 4 species on the direct line to modern humans and 5 species in total. This contrasts with current taxonomies recognising up to 23 species. The genetic proximity of humans and chimpanzees has been used to suggest these species are congeneric. Our analysis of genetic distances between them is consistent with this proposal. It is time that chimpanzees, living humans and all fossil humans be classified in Homo. The creation of new genera can no longer be a solution to the complexities of fossil morphologies. Published genetic distances between common chimpanzees and bonobos, along with evidence for interbreeding, suggest they should be assigned to a single species. The short distance between humans and chimpanzees also places a strict limit on the number of possible evolutionary 'side branches' that might be recognised on the human lineage. All fossil taxa were genetically very close to each other and likely to have been below congeneric genetic distances seen for many mammals. Our estimates of genetic divergence suggest that periods of around 2 million years are required to produce sufficient genetic distance to represent speciation. Therefore, Neanderthals and so-called H. erectus were genetically so close to contemporary H. sapiens they were unlikely to have been separate species. Thus, it is likely there was only one species of human (H. sapiens) for most of the last 2 million years. We estimate the divergence time of H. sapiens from 16 genetic distances to be around 1.7 Ma which is consistent with evidence for the earliest migration out of Africa. These findings call into question the mitochondrial "African Eve" hypothesis based on a far more recent origin for H. sapiens and show that humans did not go through a bottleneck in their recent evolutionary history. Given the large offset in evolutionary rates of molecules and morphology seen in human evolution, Homo species are likely to be characterised by high levels of morphological variation and low levels of genetic variability. Thus, molecular data suggest the limits for intraspecific morphological variation used by many palaeoanthropologists have been set too low. The role of phenotypic plasticity has been greatly underestimated in human evolution. We call into question the use of mtDNA for studies of human evolution. This DNA is under strong selection, which violates the assumption of selective neutrality. This issue should be addressed by geneticists, including a reassessment of its use for molecular clocks. There is a need for greater cooperation between palaeoanthropologists and anthropological geneticists to better understand human evolution and to bring palaeoanthropology into the mainstream of evolutionary biology. PMID- 12733396 TI - Problems with the use of cladistic analysis in palaeoanthropology. AB - Cladistic analysis is a popular method for reconstructing evolutionary relationships on the human lineage. However, it has limitations and hidden assumptions that are often not considered by palaeoanthropologists. Some researchers who are opposed to its use regard cladistics as the preferred method for taxonomic "splitters" and claim it has lead to a revitalisation of typology. Typology remains a part of human evolutionary studies, regardless of the acceptance or use of cladistics. The assumption/preference for "splitting" over "lumping" in cladistics (alpha) taxonomy and the general failure to evaluate (post-hoc) such taxonomies have served to reinforce this assertion. Researchers have also adopted a number of practices that are logically untenable or introduce considerable error. The evolutionary trend of human encephalisation, apparently isometric with body size, and concurrent reduction in the gut and masticatory apparatus, suggests continuous cladistic characters are biased by problems of body size. The method suffers a logical weakness, or circularity, leading to bias when characters with multiple states are used. Coding of such characters can only be done using prior criteria, and this is usually done using an existing phylogenetic scheme. Another problem with coding character states is the handling of variation within species. While this form of variation is usually ignored by palaeoanthropologists, when characters are recognised as varying, their treatment as a separate state adds considerable error to cladograms. The genetic proximity of humans, chimpanzees and gorillas has important implications for cladistic analyses. It is argued that chimpanzees and gorillas should be treated as ingroup taxa and an alternative outgroup such as orangutans should be used, or an (hypothetical) ancestral body plan developed. Making chimpanzees and gorillas ingroup taxa would considerably enhance the biological utility of anthropological cladograms. All published human cladograms fail to meet standard quality criteria indicating that none of them may be considered reliable. The continuing uncertainty over the number and composition of fossil human species is the largest single source of error for cladistics and human phylogenetic reconstruction. PMID- 12733397 TI - Trace element analysis of Bronze Age skeletal and crematory graves from southern Poland for diet reconstruction. AB - The basic aim of the present study was to determine the diet of human populations dating from the Bronze Age and the time of Roman influence. The osseous material under examination came from skeletal and crematory burials of the Lusatian and Przeworks Cultures, found in a cemetery at Opatow, Klobuck District, Silesian Province. Three elements: strontium, zinc and calcium were chosen as basic diet determinants. The three elements and the proportions between them are the most frequently used factors that permit a description of the relative proportion of animal and plant protein in a diet. It was assumed that the differentiation of burial ritual was paired with a diverse mode and quality of nourishment. Interdisciplinary osteological analyses, based on physicochemical studies of the odontologic material in the context of archaeological data (culture affiliation, type of burial, grave furnishings), permit a complex analysis of the issues connected with the biology of human groups, their demographic structure or, eventually, paleostratigraphy of primeval communities. It has been found that the type of burial and the richness of grave furnishings are most closely connected with the ultimate differentiation of Zn and Ca concentrations and the value of the Sr/Zn ratio. Because the richly furnished graves are at the same time mostly skeletal burials, it cannot be unequivocally stated which of the above-mentioned factors is of paramount importance. It has also been demonstrated that representatives of the Przeworsk Culture, chronologically younger than the Lusatian one that inhabited the same geographic region, show a lower Zn concentration and a higher Sr/Zn ratio. PMID- 12733398 TI - Cranial trepanation in two skulls of early medieval Italy. AB - Cranial trepanation is a practice known since prehistory in various, often geographically distant populations. In this work are presented two early medieval trepaned skulls. The first skull comes from Canosa in Apulia (6th-7th c. AD; according to radiocarbon dating), the second is from Ticineto Alessandrino (5th 8th c. AD). The skull of Canosa shows a single perforation on the bregmatic quadrant of the right parietal, whereas the Ticineto skull presents three lesions of different shapes and sizes. Two perforations are on the left parietal and the other is on the right parietal. The lesions of the Ticineto skull can be referred to a therapeutic operation following a trauma, with survival of the individual. In the Canosa skull the absence of a healing reaction suggests a pathological process, without survival of the subject. Moreover, a particular ritual practice cannot be excluded (i.e. a post-mortem trepanation). PMID- 12733400 TI - Multivariate analysis of sexual dimorphism in two types of dermatoglyphic traits in five endogamous populations of West Bengal, India. AB - Five different endogamous populations who encompass the main social rank in the caste hierarchy of West Bengal were analysed for this report. The present approach is to compare the pattern of sex differences/similarities exhibited by two different sets of dermatoglyphic traits. Cluster and discriminant analysis and Mantel test of matrix correlations were performed. The nature of variation between sexes within population groups and two types of variable sets has a good similarity in all five populations. These results strongly suggest that the two categories of dermatoglyphic variables provide similar possibilities to discriminate between the sexes in populations. PMID- 12733399 TI - Sexually dimorphic pelvic morphology in South African whites and blacks. AB - It is well known that there is metric and morphologic variation in the expression of sexual dimorphism between racial phenotypes and populations. Therefore, the purpose of this research is to assess morphologic sex differences in the pelves of South African whites and blacks. Results will be used to improve the identification of human skeletal remains by producing group specific standards. Morphologic features of both left and right os coxae were studied in a sample of 400 known sex/race individuals from the Pretoria and Dart collections. Bones that were clearly pathologic or visibly deformed were excluded from the study. Data were subjected to SPSS analysis. Results indicated that overall, public bone shape was the easiest to assess and was the most consistently reliable morphological indicator of sex in both sexes and population groups. At 88% average accuracy, the most discriminating traits in whites were pubic bone shape and subpubic concavity form. In blacks, greater sciatic notch form allowed the highest separation, averaging 87.5%, followed by pubic shape at 84.5%. Important findings included the fact that there were significant differences in the accuracy of sex determination from pelvic morphology between both males and females and whites and blacks. In conclusion, this study provides quantification of individual morphological traits in the os coxae of white and black South Africans that will be of value in forensic and archaeological analyses, especially when dealing with fragmentary remains. It also demonstrates that population differences affect the expression of sexual dimorphism and must be accounted for to develop the most effective methods of analysis. PMID- 12733401 TI - Southern Californians to drink fluoridated water. PMID- 12733403 TI - Polymer preparation instruments. New paradigm in selective dentin removal. PMID- 12733402 TI - The gentle art of periodontal maintenance. A protocol using essential oils. PMID- 12733404 TI - The very large direct resin composite restoration. PMID- 12733405 TI - No-preparation porcelain veneers. PMID- 12733406 TI - Presenting a very accurate impression procedure and testing the accuracy of the impression obtained. PMID- 12733407 TI - Optimum dental care, Part 2. The treatment phase. PMID- 12733408 TI - The heartache of separation. PMID- 12733409 TI - Anterior access endodontics. A case report. PMID- 12733410 TI - Enamel loss and occlusal vertical dimension. Causes and considerations for treatment. AB - Much of the accelerated or premature loss of vertical dimension experienced by patients is certainly preventable, and prevention should be emphasized. Recommendations on limiting any nicotine and caffeine use, supporting needs for fitness efforts, and assisted or self-management of damaging habits from stress are vital Restoratively along with aesthetics, substantial, functional vertical dimension changes may now be addressed conservatively, using a range of strategies and current state-of-the art materials. Patients should become better informed about the importance of a good dentition for their better general health. Restorative alteration of vertical dimension may not be universally essential. Nevertheless, the profession, patients, and the third-party payment community must unequivocally recognize the physiologic advantage of restoring lost FOVD. The simple replacement of missing or damaged teeth without also reviewing and often addressing related facial proportions, occlusion, and vertical dimension concerns can be too limited a treatment option. Treatment of this current and ever-increasing condition cannot be sparingly restricted to the realm of specialists. In light of current population demographics, general dentists must inform themselves of extensive, currently available information in order to improve their proficiency and gain more confidence in addressing these multifaceted, ever more common problems. Examining all treatment variables in conjunction with the proficient determination for improved masticatory function and durability will increase our potential for achieving more favorable outcomes. New research in these areas will continue to be applied in order to optimally compose and implement modern comprehensive treatment plans and better evaluate post-treatment success. Not taking the interrelated issues discussed in this article into consideration, or a failure to assess and better engage these increasingly widespread needs of our population, could severely compromise the dental health of an increasing number of current and future patients. PMID- 12733411 TI - Digital photography in your dental practice. The why's, how's, and wherefore's. PMID- 12733412 TI - Periodontal disease and diabetes mellitus. Bidirectional relationship. AB - Periodontitis is a common problem in patients with diabetes. The relationship between these 2 maladies appears bidirectional--insofar that the presence of one condition tends to promote the other, and that the meticulous management of either may assist treatment of the other. Both diabetes and periodontitis can stimulate the chronic release of proinflammatory cytokines that have a deleterious effect on periodontal tissues. The chronic systemic elevation of proinflammatory cytokines caused by periodontitis may even predispose individuals to the development of type 2 diabetes. Mechanical treatment of periodontitis (scaling and root planing), when combined with short-term administration of therapeutic levels of tetracycline-type antimicrobials, can temporarily improve glycemic control in diabetic patients, especially in those with advanced forms of periodontitis and poor glycemic control before treatment. The biochemical mechanisms suggested by these studies imply that other periodontal procedures designed to rid patients of periodontal pathogens may also improve the management of diabetes. Therefore, the authors suggest that periodontal patients with diabetes be treated in consultation with a periodontist (Figures 3a through 4b). PMID- 12733413 TI - Allergy and toxic reactions to local anesthetics. PMID- 12733414 TI - Polishing of anterior composite resin restorations. PMID- 12733415 TI - Intraoral cameras: patient education and motivation. PMID- 12733416 TI - Buyer guide to cosmetic imaging. PMID- 12733417 TI - Competency restoration: an examination of the differences between defendants predicted restorable and not restorable to competency. AB - According to the U.S. Supreme Court's decision in Jackson v. Indiana (1972), examiners must determine if a defendant has "substantial" probability of regaining competency through treatment "in the foreseeable future." Previous research has indicated that, given the low base rate of defendants unable to be restored to competency, examiners are relatively poor at predicting which defendants will regain competency. Determining the characteristics of not restorable incompetent defendants and restorable incompetent defendants is a necessary first step toward improving examiners' ability to predict a defendant's likelihood of regaining competency. This study examined the competency evaluation reports of 468 defendants evaluated for competency to stand trial. Incompetent defendants significantly differed from competent defendants with regard to age, employment status, ethnicity, criminal charges, and psychiatric diagnosis. Few significant differences existed between defendants predicted restorable and those predicted not restorable by mental health examiners--the differences that did exist were related mainly to nonpsychiatric variables. PMID- 12733418 TI - Taking responsibility for an act not committed: the influence of age and suggestibility. AB - Inherent in false confessions is a person taking responsibility for an act he or she did not commit. The risk of taking such responsibility may be elevated in juveniles. To study possible factors that influence individuals' likelihood for taking responsibility for something they did not do, participants in a laboratory experiment were led to believe they crashed a computer when in fact they had not. Participants from 3 age groups were tested: 12- and 13-year-olds, 15- and 16-year olds, and young adults. Half of the participants in each age group were presented with false evidence indicating liability. Additionally, suggestibility was investigated as a potential individual-difference factor affecting vulnerability to admissions of guilt. Results showed that younger and more suggestible participants were more likely than older and less suggestible participants to falsely take responsibility. Implications of these findings for juvenile justice are discussed. PMID- 12733419 TI - The effects of interviewer gender and child gender on the informativeness of alleged child sexual abuse victims in forensic interviews. AB - Most students of forensic interviews have focused on the interrogatory techniques used to elicit information from alleged child abuse victims. We asked how the gender of the interviewer and the gender of the child affected this process. Forensic investigators in three countries used either the NICHD structured interview protocol or local standard interview practices to interview 672 alleged victims who ranged in age from 4 to 14 years. Analyses of the interviews showed significant effects of gender on both the interviewers' behavior and the amount of information provided by children. Female interviewers asked boys more invitations, as well as absolutely and proportionally more suggestive questions, than they did girls, whereas male interviewers interviewed boys and girls similarly. Children's responses varied depending on their gender and age, the gender of the interviewer, and the type of question asked. Girls of all ages provided more information in response to directive questions posed by female rather than male interviewers whereas boys did not respond differently to male and female interviewers. The oldest girls provided more information in response to option-posing questions posed by male interviewers. More information was provided by the younger children in response to suggestive prompts from interviewers of the opposite gender. The gender-of-interviewer effects were attenuated in protocol-guided interviews. PMID- 12733420 TI - Punishment goals of crime victims. AB - Research on subjective punishment goals has focused on the perspective of third party observers of criminal offenses and neglected the perspective of victims. This study investigates punishment goals among 174 adult crime victims (rape and nonsexual assault) for each participant's real criminal case. Scales measuring support for punishment goals are constructed by factor analysis of an 18-item list. Results show that 5 highly supported goals can be distinguished: retaliation, recognition of victim status, confirmation of societal values, victim security, and societal security. Analysis of relations between punishment goal scales and personal variables, situational variables, and demanded punishment severity corroborates the view that the punishment goals revealed can be classified according to the two independent dichotomies of moral versus instrumental goals, and micro versus macro goals. PMID- 12733421 TI - Behavioral confirmation in the interrogation room: on the dangers of presuming guilt. AB - A two-phased experiment tested the hypothesis that the presumption of guilt that underlies police interrogations activates a process of behavioral confirmation. In Phase I, 52 suspects guilty or innocent of a mock theft were questioned by 52 interrogators led to believe that most suspects were guilty or innocent. Interrogators armed with guilty as opposed to innocent expectations selected more guilt-presumptive questions, used more interrogation techniques, judged the suspect to be guilty, and exerted more pressure to get a confession--particularly when paired with innocent suspects. In Phase II, neutral observers listened to audiotapes of the suspect, interrogator, or both. They perceived suspects in the guilty expectations condition as more defensive--and as somewhat more guilty. Results indicate that a presumption of guilt sets in motion a process of behavioral confirmation by which expectations influence the interrogator's behavior, the suspect's behavior, and ultimately the judgments of neutral observers. PMID- 12733422 TI - Jurors' perceptions of adolescent sexual assault victims who have intellectual disabilities. AB - Children and adolescents with intellectual disabilities are especially likely to be sexually abused. Even so, their claims are not likely to be heard in court, possibly because people assume that jurors will not believe them. We tested this assumption in a mock-trial study in which 160 men and women watched videotaped excerpts from an actual trial. As predicted, when the 16-year-old sexual assault victim was portrayed as "mildly mentally retarded" instead of as "having average intelligence," jurors were more likely to vote guilty and had more confidence in the defendant's guilt; considered the victim to be more credible and the defendant to be less credible as witnesses; and rated the victim as more honest, less capable of fabricating the sexual abuse accusation, and less likely to have fabricated the sexual abuse accusation. Men and women were affected similarly by the disability manipulation, but women were generally more pro-prosecution in their case judgments and perceptions than were men. Finally, jurors who had more liberal views toward persons with disabilities were more likely than other jurors to make pro-prosecution judgments on measures of guilt. Implications for psychological theory and the law are discussed. PMID- 12733424 TI - Hereditary Creutzfeldt-Jakob disease and fatal familial insomnia. AB - Studies on hereditary CJD and FFI have contributed greatly to the understanding of all forms of prion disease. Most importantly, they have provided strong support for the prion hypothesis [2]. The linkage of pathogenic PRNP mutations to human prion disease strengthens the notion that a change in PrP conformation is a key event that triggers the development of the disease. Although hereditary CJD and FFI account for only 10% of all cases of human prion disease, they provide a unique opportunity for studying disease pathogenesis initiated by perturbation in the PrP structure. An understanding of the events that accompany a change in PrP conformation has far-reaching implications for sCJD (the most common form of the disease) and for sporadic fatal insomnia. A wealth of available evidence indicates that a common pathway in disease pathogenesis may be shared by both the sporadic and the hereditary forms of prion disease, except that the initiating events are stochastic in the former, rather than predetermined by the presence of a germ-line mutation. In addition, investigations of hereditary CJD and FFI have provided plausible mechanisms of phenotypic heterogeneity in prion disease, a phenomenon analogous to the "prion strain" diversity in animal prion disease. Although many other neurodegenerative diseases such as Alzheimer's disease, amyotrophic lateral sclerosis, and Huntington's chorea are fairly homogeneous in disease phenotype, prion disease includes many clinically and pathologically distinct disease entities. In hereditary prion disease, the disease phenotype is likely to be determined by the combined effect of pathogenic mutations, codon 129 polymorphism, and the type of PrPSc. The pathogenic mutations include point mutations that are located mostly in the central and C-terminal region of PrP, and deletion and insertion mutations that are located in the N-terminal region. It is conceivable that these distinct types of mutations may result in differential changes in conformation or stability of PrP. The codon 129 polymorphism plays a twofold role in modulating the disease outcome. On the mutant allele, it determines the basic features of the disease phenotype--as in the case of FFI and CJD178--that result respectively from the coupling of M or V at codon 129 with the D178N mutation. On the normal allele, it may modulate the severity of the phenotype. A PrPSc subtype is encoded by the PRNP haplotype, and subsequently is generated by a conformational conversion process that transforms the cellular isoform to the pathogenic protein. The site for the formation of a specific PrPSc conformer and its accumulation in different brain regions are likely to contribute to the clinical features and pathologic lesions. The phenotypic homogeneity in other neurologic diseases, including Alzheimer's disease, may be due, in part, to the lack of a powerful genetic modifier such as the codon 129 polymorphism in the PrP gene, and the lack of the ability of affected gene products such as PrP to assume multiple protein conformations. Clearly, the remaining issue in the understanding of pathogenesis of prion disease is a detailed and accurate knowledge of the in vivo processes and conditions for the formation of PrPSc that inevitably lead to the development and expression of the disease. This knowledge will enable the development of a rational and effective strategy for therapeutic intervention. PMID- 12733423 TI - Perspectives on prion biology, prion disease pathogenesis, and pharmacologic approaches to treatment. AB - The main goals of this article have been to summarize our current understanding of the biology of PrP, the propagation of prions, and the etiology and pathogenesis of each form of prion disease (familial, sporadic, and infectious); and to review current rational pharmacologic strategies for treatment of prion diseases. Each of these subjects is presented primarily from the perspective of investigations performed by the prion disease research laboratories at the University of California in San Francisco and by its many collaborators in the United States and abroad. This review focuses on key results from the hundreds of transgenic mouse lines expressing different PrP constructs that have been used to determine the roles played by different PrPSc and PrPC domains in prion propagation and the prion disease phenotype. PMID- 12733427 TI - Diagnosis of prion diseases. AB - This article reviews the various techniques and methodologies for the recognition and diagnosis of clinical and neuropathologic variants of human prion diseases. PMID- 12733425 TI - Hereditary prion protein amyloidoses. AB - Prion protein (PrP) amyloid accumulation is the pathologic hallmark of some inherited prion diseases such as Gerstmann-Straussler-Scheinker disease (GSS) and PrP cerebral amyloid angiopathy (PrP-CAA). In GSS, parenchymal amyloidosis may coexist with spongiform degeneration or neurofibrillary tangles, whereas in PrP CAA, vascular amyloid coexists with neurofibrillary tangles. In GSS, N-truncated and C-truncated proteinase K-resistant PrP isoforms are present in the brain. PMID- 12733426 TI - Variant Creutzfeldt-Jakob disease. AB - Variant CJD is a novel human prion disease that represents the first known occasion in which animal prion diseases have been transmitted to humans. There are many uncertainties concerning vCJD, including the mechanism of transmission between species, the extent of human exposure to the BSE agent, the infectious dose for humans, and the future burden of human disease. It is hoped that continuing scientific research may lead to answers to some of these questions and that further understanding of the mechanism of prion replication may lead to the development of effective treatment. Indeed a recent publication has suggested that the drugs quinacrine or chloropromazine may be candidates for the treatment of human prion diseases [42]. PMID- 12733428 TI - Transmissible spongiform encephalopathy update and implications for blood safety. AB - At this time, the accumulated evidence does not support the implementation of measures targeted against the risk of transfusion transmission of sporadic, familial, or iatrogenic CJD. Evolving information about vCJD, however, suggests that policy makers need to consider implementing measures to protect against exposure to vCJD, if such measures themselves do not lead to decreased blood safety. Surveillance of human TSEs and investigation of the risk of transfusion transmission must continue in order to provide further refinements in blood safety policy. PMID- 12733430 TI - A gene-targeted mouse model of P102L Gerstmann-Straussler-Scheinker syndrome. AB - Transgenic mice that contain a proline to leucine mutation at amino acid 101 in the endogenous murine PrP gene have been produced by gene targeting. This line of mice was generated to model the mutation thought to be responsible for P102L GSS, a familial TSE disease in humans. Genetargeted 101LL mice showed no evidence of spontaneous TSE disease in their lifetime and were unable to transmit any neurologic disease to other 101LL transgenic mice. 101LL mice have, however, been shown to demonstrate altered susceptibility to several TSE strains, and have shown reduced incubation times with TSE agents that do not readily transmit to wild-type mice. The 101L mutation does not appear to destabilize PrP and promote conversion to PrPSc, because incubation times are increased with mouse-passaged TSE strains and vCJD. PrPSc also can be difficult to detect in 101LL mice infected with some TSE strains. We, therefore, have been unable to substantiate the existence of either genetic disease or infectious PrP with the P101L transgenic model, but have provided evidence of altered incubation times of TSE disease in mice carrying the 101L mutation in their PrP protein. We also have shown that mutations in the N-terminal region of PrP can have a major influence over both incubation time and targeting of TSE disease. PMID- 12733429 TI - Scrapie and chronic wasting disease. AB - Scrapie and CWD share many features. There are marked similarities in the clinical presentations, the lesions, and the pathogenesis of these diseases, and some similarities in the epidemiology. Extrapolation from the scrapie model of TSE disease to CWD--which occurs in three different species, and should not be considered to be uniform in their response--may be erroneous, however. Such differences may influence diagnostics (e.g., the amount and distribution of PrPC in these different species), pathogenesis (e.g., the influence of genetics on susceptibility and resistance), and epidemiology (e.g., the mode and dynamics of transmission and influences of domestication). IHC is used widely for diagnostics and in the study of the pathogenesis of scrapie and CWD. This technique holds promise for antemortem diagnosis of infection in the peripheral lymphoid tissues such as lymphoid follicles of the nictitating membrane and the tonsil. PMID- 12733431 TI - A murine model of a familial prion disease. AB - We have produced a mouse model of a familial prion disorder by introduction of a transgene that encodes the moPrP homolog of a nine-octapeptide insertional mutant associated with an inherited form of CJD in humans. These mice develop progressive neurologic symptoms, display neuropathologic changes, and accumulate a form of mutant PrP in their brains and peripheral tissues that displays some of the biochemical properties of PrPSc. These mice have been extremely valuable for analyzing the cellular and biochemical mechanisms involved in inherited prion disorders and correlating the appearance of the PrPSc-like form with clinical and neuropathologic findings. Because the mutant protein in the mice is highly neurotoxic but appears to lack infectivity, further analysis of its properties promises to shed new light on the molecular distinction between pathogenic and infectious forms of PrP. PMID- 12733432 TI - Therapeutic approaches to prion diseases. AB - Prion diseases are unique in that they comprise sporadic, genetic, and iatrogenically or environmentally acquired forms. When disease is acquired by peripheral route, neuroinvasion occurs via at least two different neural pathways (vague and splanchnic nerves) and is usually preceded by prion propagation in secondary lymphoid organs. Conversely, in the other etiologic forms, PrPSc formation occurs within, and is apparently limited to, the CNS. Longitudinal studies on experimental scrapie indicate that substantial neuropathologic changes (i.e., glial activation and nerve cell degeneration) already are present before the onset of symptoms and are topographically related to PrPSc deposits. Accordingly, any effective intervention should start during the preclinical stage of disease, and be aimed at preventing neuroinvasion or PrPSc propagation in the CNS. Unfortunately, no tests are available currently to detect presymptomatic individuals, except for carriers of pathogenic mutations of the PRNP gene. Inhibition of PrPSc formation can be achieved through (1) abrogation of PrPC synthesis or prevention of its transport to the cell surface; (2) stabilization of the PrPC structure to make its conformational change unfavorable; (3) sequestration of PrPSc; (4) reversion of PrPSc to a protease-sensitive form; or (5) interference with the interaction between PrPC, PrPSc, and other macromolecules that feature in the conversion process. The compounds that have some effectiveness in in vitro, cell culture, or animal models of prion disease seem to operate through one of these mechanisms (see Table 1); however, even the most effective drugs only work when administered at the time of infection or very short thereafter, and these conditions are incurable at present. The heterogeneity and complexity of the etiopathogenesis of prion diseases suggest that various strategies and a combination of several compounds with different modes of actions are likely necessary for prevention and treatment. Major efforts should be focused on the development of preclinical diagnostic tests in conjunction with immunization strategies for diseases acquired by peripheral route and identification of more effective compounds for the other etiological forms. PMID- 12733433 TI - Three-dimensional structures of the prion protein and its doppel. AB - This article discussed the implications of the structures of PrP and Dpl--with their unusual folds containing N-terminal flexible tails and C-terminal globular domains--to the physiologic functions of PrPC and Dpl, and investigations of a possible structural basis of familial human TSEs. Further relations between TSEs and the PrP structure would include the species barrier of TSEs (which seems to be associated with species-specific structural characteristics of PrPC [25,39,67]), and the conformational transition from PrPC to PrPSc using, for example, molecular dynamic simulations [68,69]. Due to the lack of knowledge on physiologic functions of PrPC, however, and the remaining uncertainty about the exact role of the PrP in TSE pathology, it appears that most or all of the physiologically relevant structure-function correlations of PrPC have yet to be identified. PMID- 12733435 TI - [Proceedings of the First Puerto Rican Public Health Conference. 10-12 April 2002, San Juan, Puerto Rico]. PMID- 12733434 TI - In vitro conversion of normal prion protein into pathologic isoforms. AB - The in vitro conversion techniques in cell-free and cell culture systems have provided tools to adequately study the underlying mechanism of TSEs, namely PrP conversion. These systems also have provided tools that make it easier to study the interspecies and intraspecies transmissibilities of TSEs. Finally, these systems also may assist in the discovery of TSE therapeutic strategies and in the development of extremely sensitive TSE detection techniques. In vivo TSE transmission studies are limited to (transgenic) animals (mostly mice). Although the cell culture systems also are restricted in their species-range (mostly mouse), the currently used cell-free systems. Allow studying almost all possible species barriers (including the potential transmission of various TSEs to humans). One advantage of the cell culture systems, however, is that they generate do novo TSE infectivity. Studies using cell cultures also take into account several cofactors in addition to PrP that might be involved in replication the TSE agent. Although the in vitro systems provide accurate tools to study TSE agent parameters, they mainly or only focus on the molecular processes of PrP conversion. Other factors (i.e., host genetic factors [99]) that, for example, determine the differential uptake of the TSE agent from the environment, might play an additional role in determining the susceptibility of hosts for TSEs and on the transmission of the disease among individuals. PMID- 12733436 TI - [The future of public health: different perspectives and focuses]. PMID- 12733437 TI - [Challenges in public health for the new century]. AB - The history of public health has followed different trends, depending on the prevailing theories of disease causality and the interventions that these suggest. From the concern with miasmas to the germ theory, to the stress on lifestyles and the current interest on the linkages between health, economic welfare, and social status, the proposed policies are contingent on how health and disease are conceptualized and explained. Ironically, now that there is increasing evidence on the socio-economic determinants of health, interventions continue to focus on specific target groups or disease categories, as well as on the medical sector. PMID- 12733438 TI - [The ethics of public health]. PMID- 12733439 TI - [Ethics in epidemiological research]. PMID- 12733440 TI - [Demographic ethics]. PMID- 12733441 TI - [The necessity of incorporating ethics into the formulation of public environmental policy]. PMID- 12733442 TI - [Ethics in health services administration]. PMID- 12733443 TI - [Social participation and community empowerment in public health]. PMID- 12733444 TI - [The woman who emigrates to Puerto Rico: demographic and legal considerations]. PMID- 12733445 TI - [War on Drugs or War against Health? The pitfalls for public health of Puerto Rican drug policy]. AB - Puerto Rico has followed the United States in adopting drug policy sustained on a criminal justice model that limits the opportunities to address problematic drug use through public health interventions. Demand for illegal drugs is controlled by criminalizing drug use and applying jail sentences for drug offenses. These strategies marginalize drug users and reduce opportunities to minimize health risks applying public health measures. Production and sale of illegal drugs is criminalized with the intent of dissuading drug use, with adverse unintended health effects that impact both drug users and non-drug users in the community. The present work reviews the assumptions of the punitive prohibitionist model and its outcomes that present themselves as public health challenges in Puerto Rico. It also presents those principles that should sustain pragmatic drug policy to address problematic drug use from a health and social perspective. PMID- 12733446 TI - An overview of reported youth violence in Puerto Rico. AB - OBJECTIVES: The purpose of this paper is to describe the prevalence of school and youth violence in Puerto Rico based on several representative samples surveyed in the Island during the 1990's. BACKGROUND: Reviewing the available data on youth violence is necessary in order to decide the best strategies to follow in developing new ways to prevent and monitor it. METHODS: Three surveys were reviewed in this paper: The Youth Risk Behavior Survey conducted by the Center for Disease Control (1991, 1995, 1997); The Methods for the Epidemiology of Child and Adolescent Mental Disorders Study (1991); and the Youth Survey (1997-98). RESULTS: It was found that the prevalence of reported school violence in Puerto Rico tends to be somewhat lower than the prevalence reported in other sites of the U.S.A. Age and gender has a dramatic influence in the reports of youth violence in Puerto Rico. Male students are three times as likely to carry weapons than females and almost twice as likely to report hurting someone than females. Less than 10% of the male students ages 12-13 report carrying a weapon while fully 30% of those 18 year olds reported carrying a weapon. Being arrested also increases from only 3% in the youngest students surveyed to 10% on the oldest group (18 or older). CONCLUSION: The CDC has recently reported that since the early 90's there has been a decrease in reported school violence in the U.S.A. This analysis of youth surveys replicated a similar tendency for Puerto Rican youngsters. Implications for prevention programs are discussed. PMID- 12733447 TI - [The identification of risk factors and protective factors for the prevention of youth violence: toward a community strategic plan]. PMID- 12733448 TI - [The social construction of violence in a group of young people of both sexes in a public middle school]. AB - In this article we will address some factors that promote violence in schools from the youth viewpoint. We present an overview of what young people understand about violence; in which way its take places; in what forms they express it; and what aspects they consider more important than others. At the same time, it compares the social construction of violence on both, boys and girls, and describe how violence is interpreted from the gender perspective. The latter means that assumption of violent behavior varies with the gender of the person describing it. In other words, any project that pretends to promote schools free of violence need to deal with gender issues. Finally, I will share a few general considerations about what we may call social construction of violence in young people and its implications for a violence prevention project. PMID- 12733449 TI - Developing a center for Hispanic youth violence prevention. AB - Youth violence is a serious public health problem, described by the Surgeon General's Report on Youth Violence (2001). Homicide is the second cause of death for the 15-24 age population in the USA and Puerto Rico; intentional injuries also result in significant morbidity. Youth are at greater risk of being victims than perpetrators of violence. African American and Hispanic youth are at particular risk. Effective, community-based initiatives to prevent youth violence are necessary to interrupt the cycle of violence. In 2000, the Centers for Disease Control and Prevention of the Department of Health and Human Services awarded grants to establish ten Academic Centers of Excellence for Youth Violence Prevention. This article describes the development of the Center for Hispanic Youth Violence Prevention, of the FILIUS Institute of the University of Puerto Rico. PMID- 12733450 TI - [Public health activism]. PMID- 12733451 TI - [Vieques and activism]. PMID- 12733452 TI - [Mexican health reform]. PMID- 12733453 TI - Analysis and modeling of human postural control. PMID- 12733455 TI - Changes in postural control with aging and Parkinson's disease. PMID- 12733454 TI - Change-in-support reactions for balance recovery. PMID- 12733456 TI - Gaining insight by going in circles. PMID- 12733458 TI - Strategies for dynamic stability during adaptive human locomotion. PMID- 12733457 TI - Directional aspects of balance corrections in man. PMID- 12733459 TI - Adaptation to vibratory perturbations in postural control. PMID- 12733460 TI - Changes in walking strategies after spaceflight. PMID- 12733462 TI - Nonstationarities of postural sway. PMID- 12733461 TI - Simplifying the complexities of maintaining balance. PMID- 12733463 TI - Noise-enhanced human sensorimotor function. PMID- 12733464 TI - Balance prostheses for postural control. PMID- 12733465 TI - The future of clinical engineering: the challenge of change. AB - Clinical engineering is at a strategic inflection point. Technical, economic, regulatory, and cultural dynamics are at work shaping the future of healthcare delivery. As the nature of healthcare delivery is transformed by these forces, the types and mix of technology management and support services needed by the industry are changing significantly. Clinical engineering has a relatively short opportunity to adopt a service model that will meet these changing needs. Delay or failure to adopt an effective service model as we pass through the inflection point will result in a diminished role for clinical engineering in healthcare technology management as other technical professionals move in to fill the need. The question is: will clinical engineering rise to the challenge? PMID- 12733466 TI - More bad news about money and medical science. PMID- 12733467 TI - Cell signaling--is it new? PMID- 12733468 TI - Reading the fine print of the human genome. PMID- 12733469 TI - Design controls. PMID- 12733471 TI - Improving communication of drug risks to prevent patient injury: proceedings of a workshop. AB - PURPOSE: The Centers for Education & Research on Therapeutics (CERTs) is conducting a series of workshops on managing the risks of therapeutics, with the ultimate goal to develop an agenda for research and education about risk and its management. This paper presents the results of the first workshop in the series, a 2-day meeting focused on communication of drug risks to healthcare professionals and patients. METHODS: The 50 workshop participants represented the medical-products industry, academia, consumer groups, regulatory bodies and the media. Together, they sought to identify and understand barriers to successful risk communication, to identify tools or methods that could improve risk communication, and to develop research and education agendas that would lead to better risk communication in the future. RESULTS: Limitations of current methods of risk communication were identified, and research and education agendas were proposed to clarify and resolve these issues. CONCLUSION: Common themes for potential solutions include enhanced education of healthcare providers, increased motivation of patients and families, use of creative communication technologies, and better organization of and access to medical records and information. PMID- 12733470 TI - Increased US prescription trends associated with the CDC Bacillus anthracis antimicrobial postexposure prophylaxis campaign. AB - PURPOSE: We evaluated national outpatient antimicrobial prescription trends in relation to the first United States case of inhalational anthrax due to the intentional delivery of Bacillus anthracis (B. anthracis) spores. METHODS: We queried IMS HEALTH's National Prescription Audit Plus7 database for two 6-month periods (July-December) in 2001 and 2000 to describe outpatient prescription trends of antimicrobials recommended during the Centers for Disease Control and Prevention's (CDC) postexposure prophylaxis campaign. RESULTS: Overall, antimicrobial utilization for the referent 6-month time frame was greater in 2000 compared to 2001. In contrast, ciprofloxacin utilization was greater in 2001 during October, the month following the index case, increasing by more than 40% over utilization in October 2000. Similarly, doxycycline utilization increased by 30% during October/November. This corresponded to relative increases in US utilization for ciprofloxacin of approximately 160,000 prescriptions for the month of October and for doxycycline of approximately 96,000 prescriptions during October and 120,000 prescriptions for November. CONCLUSIONS: We conclude more widespread prescribing of ciprofloxacin and doxycycline occurred in response to the first US bioterrorist-associated anthrax attacks than was warranted based upon confirmed or suspected B. anthracis exposure alone. PMID- 12733472 TI - Risk of anaphylaxis in a hospital population in relation to the use of various drugs: an international study. AB - PURPOSE: To evaluate the risk of anaphylaxis following exposure to medications given during hospitalization. METHODS: Cases with anaphylaxis and controls, matched according to length of hospital stay, were enrolled from hospitals in Hungary, Spain, India and Sweden. Exposures were recorded from the hospital record. The incidence of anaphylaxis was calculated among those exposed to various drugs, with denominators estimated from exposure rates in the controls and number of hospitalizations. RESULTS: Among 184 cases and 1003 controls, the incidence of anaphylaxis was in the range of 5-15 cases per 100,000 exposed patients for most analgesics and antibiotics administered orally or parenterally; for parenteral penicillin, it was 32. Incidence estimates were somewhat higher for blood, dextran, pentoxifylline and both ionic and nonionic contrast media, ranging from 35 to 95 cases. The highest estimates were for streptokinase and plasma, at 378 and 284. With no exposed controls, the incidence could not be calculated for anti-snake venom, but it was clearly higher than for other exposures. CONCLUSIONS: Although anaphylaxis is known to occur following exposure to a large number of drugs, the incidence is not well documented. We have demonstrated a relatively low risk for dipyrone, diclofenac, paracetamol, ampicillin, cloxacillin and cephalosporins. An intermediate risk was shown for parenteral penicillin, dextran, contrast media, blood and pentoxifylline. The highest incidence was observed for plasma, streptokinase and anti-snake venom (the latter used only in India). PMID- 12733473 TI - Antihypertensive drug-associated sexual dysfunction: a prescription analysis based study. AB - PURPOSE: To determine whether primary care physicians take into consideration age, gender and diabetes mellitus as risk factors for sexual dysfunction (SD) when prescribing antihypertensives. METHODS: A prescribing survey on hyperternsive patients with or without diabetes mellitus in primary care setting of Bahrain was carried out. RESULTS: In 5301 hypertensive patients, we found that a beta-blocker (atenolol) was the most commonly prescribed drug and was significantly more often prescribed for young male hypertensives (< 45 years). A relatively high dose of atenolol (100 mg/d) was extensively used as both mono- and combination-therapies. With the exception of middle-aged hypertensives (45-64 years) and elderly diabetic-hypertensives (> or = 65 years), no significant age- and gender-related differences were observed regarding the prescription of thiazide diuretics. A significant trend of prescribing methyldopa in females as monotherapy was evident. Angiotensin converting enzyme (ACE) inhibitors, the second most commonly prescribed drugs, were significantly more often prescribed for young, middle-aged hypertensive males and for middle-aged diabetic hypertensive males. beta-blocker/diuretic combination was prescribed least for the young and elderly hypertensive males. However, prescription of an ACE inhibitor with either a diuretic or a calcium channel blocker (CCBs) was less frequent and lacked gender-related differences. CONCLUSIONS: We infer that antihypertensive-induced SD received little attention as was evident from: (a) over-prescription of relatively high dose of beta-blockers to young hypertensive males; (b) lack of gender- and age-related difference regarding the type and dose of diuretics prescribed; (c) underutilization of effective and complementary combinations expected to cause least impact on sexual function, such as ACE inhibitors with either a diuretic or with a CCB; and (d) a relative lack of discrimination while selecting antihypertensive drugs, particularly as monotherapy between hypertensive versus diabetic-hypertensive patients. With the exception of ACE inhibitors alone and combination of diuretic/beta-blocker, the choice of antihypertensives did not conform to international guidelines. In view of drug-induced SD adversely influencing the quality of life and thereby drug compliance, interventions aimed at improving physicians' awareness should be attempted. PMID- 12733474 TI - Antibiotic prescribing for otitis media: how well does it match published guidelines? AB - Otitis media (OM) accounts for approximately 31 million physician visits annually, and is typically treated with antibiotics. Amoxicillin is the recommended first-line treatment; appropriate antibiotic use slows the development of resistance. We analyzed insurance claims from families of employees working at a single company to determine if OM treatments (1) matched published standards; and (2) varied by patient characteristics, type of physician practice or geographic location. Persons diagnosed with OM between 1996 and 1999 were matched to prescription drug claims for those who filled an antibiotic prescription within 3 days of diagnosis. Physicians prescribed amoxicillin for only 31% of acute cases and 19% of recurrent cases. For acute infections in children < or = 2 years the prescribed duration often matched the standard of 10 days, but for persons > 2 years the prescribed duration was often longer than the suggested duration of 5-7 days. For persons of all ages with recurrent infections, the prescribed duration was often shorter than suggested (10 days versus > or = 14 days). There were only modest variations by urban/rural location or provider type. PMID- 12733475 TI - A study of drug use during pregnancy in a teaching hospital in western Nepal. AB - Information on the use of drugs during pregnancy is scarce and rather anecdotal. Careful consideration of the benefit to the mother and the risk to the fetus is required when prescribing drugs during pregnancy. The aim of this study was to gain knowledge on this issue in western Nepal. 2156 prescriptions of pregnant women were collected at random from the antenatal care (ANC) in obstetrics out patient department (OPD) at Manipal Teaching Hospital (MTH), Nepal and analyzed for this study. The mean maternal age and hemoglobin concentration were 25 years and 12.21 g/dl, respectively. Twenty-three percent women attended obstetric OPD due to maternal disorders other than routine ANC (77%). Problem oriented drug use was due to nausea/vomiting (4.7%), dyspepsia (3.1%), and per vaginal spotting/bleeding (3.4%), mainly. Most of the women got 2-3 drugs and commonly included nutritional supplementation and tetanus toxoid. The average number of drugs/prescription was 2.00, 15.37% and 64.8% drugs were prescribed by generic name and as fixed dose combinations, respectively. The most commonly prescribed drugs were nutritional supplements like iron, folate, calcium, vitamins (72.8%), followed by tetanus toxoid (12.4%), gastrointestinals (5%), antimicrobials (4.6%), etc. Though, the selection of drugs was rational in most of the cases, some anomalies were observed and discussed with the clinicians. Our data reflect the general extent and prescribing pattern for those Nepalese pregnant women attending hospital in western Nepal. PMID- 12733476 TI - Trends in the use of antituberculosis drugs in Spain 1993-1998. AB - PURPOSE: To determine and analyse the trends in the use of antituberculosis agents in Spain during 1993-1998. METHODS: Pharmacoepidemiological study on the use of antituberculosis drugs using the data from prescriptions redeemed by the National Health Service. RESULTS: The use of antituberculosis drugs over the study period was 6,271,909 defined daily doses (DDDs) in 1993, which fell to 4,433,066 DDDs in 1998. A decreasing trend was thus seen, involving a 29.6% drop between the beginning and the end of the study period. The use of daily doses per 1000 inhabitants was 0.42 in 1993 and 0.30 in the end of the period in 1998. The drug most used during the period of study was Rifinah. In 1993, its percentage of use was 41.2, which then decreased to 34.5 in 1998. After this drug, Rifampicin and Etambutol were those with the highest percentages of use, with values close to 18 and 15, respectively. Regarding estimation of tuberculosis, the prevalence was 25.25 cases per 100,000 inhabitants in 1993 and 16.62 in 1998. A decrease of 23.1% occurred in the cost in pesetas between 1993 and 1998. CONCLUSIONS: Drug's combinations are the most common therapy for tuberculosis and there were important differences between regions. The highest estimation of this disease is in the northwest region of Spain. PMID- 12733477 TI - Relative risks of reported serious injury and death associated with hemostasis devices by gender. AB - PURPOSE: To assess relative risks by gender of reported serious injuries and deaths associated with the use of hemostasis devices, stratified by year of report, type of injury, and type of device. METHODS: Reports from the Food and Drug Administration's Medical Device Reporting system and National Center for Health Statistics data on use of cardiac catheterization were used to estimate relative risks of reported serious injuries and deaths by gender. RESULTS: Estimated risks of reported serious injuries and deaths associated with hemostasis devices were two to three times greater in females than in males for hemorrhage and hematoma (p < 0.0001), but there was no significant difference in risks by gender for infection. CONCLUSIONS: Cardiac catheterization is sometimes associated with serious injuries and deaths. Among patients who receive hemostasis devices, the risk of these events are disproportionately greater in women. PMID- 12733478 TI - Trends in antimicrobial utilization at a Spanish general hospital during a 5-year period. AB - PURPOSE: Antimicrobials are a major part of hospital pharmacy budgets and must be considered in resource planning and spending projections. This study describes the profile of antibiotic use at a medium-sized hospital (by examining the ICU separately) and analyses its evolution over the period 1996-2000. METHODS: Descriptive and retrospective study. Pharmacy records were reviewed to identify oral and parenteral antimicrobial agents administered to inpatients. Results were expressed in Daily Defined Doses (DDD) per 100 stays and day. RESULTS: During the 5-year study period 176.162 DDD/100 s-d of antibiotics were consumed in the ICU, whereas in the rest of the hospital usage was much lower (54.438 DDD/100 s-d). Aminoglycosides, cephalosporins, penicillins, glycopeptides and carbapenems were the most commonly used groups of antimicrobials in the ICU, and penicillins, cephalosporins, trimethoprim/sulfonamide combinations, aminoglycosides and quinolones in the rest of the hospital. CONCLUSIONS: ICUs have some special features which make them different to the other inpatient areas. Because of that fact we consider it important to study this specific patient-care area separately. PMID- 12733480 TI - Pharmacoepidemiology and drug safety. PMID- 12733479 TI - The effect of backgrounds in safety analysis: the impact of comparison cases on what you see. PMID- 12733481 TI - Steps to increase the profitability of your practice. AB - Many physicians feel high levels of frustration and anxiety in the face of a financial picture that seems out of their control. However, there are ways physicians can gain back some control by improving both productivity and efficiency. First, examine your physical facility to determine where improvements in work flow and communications can be made. Next, save time by effectively delegating clinical and administrative tasks that don't require your expertise or skills. Improve productivity and patient flow by setting target numbers for patient visits and number of work hours. Finally, don't forget to recognize the value of your staff to create a higher level of success. PMID- 12733483 TI - Office space planning and design for medical practices, Part 1: An overview. AB - The physical environment in which physicians practice is important to their professional and financial success. Providing a pleasant and efficient workspace enhances morale for both staff and patients and increases practice prestige, efficiency of operations, and ultimately revenue and profitability. This series of articles details the questions that need to be considered in assessing office space requirements. This assessment is critical to form the decision to reconfigure existing space or build anew. PMID- 12733482 TI - Can the doctor still see me? What happens when patients arrive late? AB - Demands to optimize productivity and quality require a patient scheduling system that can balance patient demand and clinic resources. The consequences of unscheduled and late patient arrivals on operational efficiencies have been documented. Less understood is the impact of unscheduled and late arrivals on the quality of service each receives. This article examines the impact of unscheduled and late patient arrivals on operational, clinical and administrative outcomes that affect quality of care of children potentially eligible for immunizations. An unexpected finding was the generally better and faster levels of service for late arrivals. PMID- 12733484 TI - Wireless computing and health care. AB - Wireless computing technology has progressed sufficiently that it is now being incorporated into health care communications. These advances are linked to major improvements in general computer capabilities, the introduction of global satellite systems, and changes in government regulation of airwave access. This article describes some of these advances and their application in health care settings. PMID- 12733485 TI - Deploying the EMR, Part 1: The basic needs. AB - This case study presents a collection of principles and guidelines regarding the process of selecting and deploying an electronic medical record (EMR) in a physician's practice. Practical advice and observations are detailed about what was learned and how the practice has benefited. In Part 1 of this article, the authors identify the functions that are crucial in an ideal EMR. In Part 2, they will provide their guidelines for implementing these functions. PMID- 12733486 TI - The practice of forensic medicine: opportunity with strings attached. AB - Contrary to popular belief, forensic medicine is not limited to grisly anatomic pathology, or even to criminal cases. It is better defined as practice at any of the many interfaces of medicine and law. This article will discuss some topics and procedures of forensic practice that are shared by many forms of forensic work, regardless of specialty. It will focus on private consultation, in which the physician provides expertise to lawyers, courts, or organizations involved in legal or administrative matters. PMID- 12733487 TI - Outsourcing your insurance collection follow-up: making the right business decision. PMID- 12733488 TI - Creating job descriptions and a job applicant wish list. AB - How can you recognize the best job applicant for an open position in your practice when he or she shows up at your door? The up-to-date, comprehensive job description will be your best guide. In this article, the author suggests what information should be included in job descriptions, who should write job descriptions for your practice, and when and how to determine appropriate salary and benefits for each job description. In addition, this article offers an important disclaimer for job descriptions that will protect you legally and that every employee in your practice should sign. PMID- 12733490 TI - [Ophthalmology publishing]. PMID- 12733489 TI - First proficiency testing to evaluate the ability of European Union National Reference Laboratories to detect staphylococcal enterotoxins in milk products. AB - The European Commission has designed a network of European Union-National Reference Laboratories (EU-NRLs), coordinated by a Community Reference Laboratory (CRL), for control of hygiene of milk and milk products (Council Directive 92/46/ECC). As a common contaminant of milk and milk products such as cheese, staphylococcal enterotoxins are often involved in human outbreaks and should be monitored regularly. The main tasks of the EU-CRLs were to select and transfer to the EU-NRLs a reference method for detection of enterotoxins, and to set up proficiency testing to evaluate the competency of the European laboratory network. The first interlaboratory exercise was performed on samples of freeze dried cheese inoculated with 2 levels of staphylococcal enterotoxins (0.1 and 0.25 ng/g) and on an uninoculated control. These levels were chosen considering the EU regulation for staphylococcal enterotoxins in milk and milk products and the limit of detection of the enzyme-linked immunosorbent assay test recommended in the reference method. The trial was conducted according to the recommendations of ISO Guide 43. Results produced by laboratories were compiled and compared through statistical analysis. Except for data from 2 laboratories for the uninoculated control and cheese inoculated at 0.1 ng/g, all laboratories produced satisfactory results, showing the ability of the EU-NRL network to monitor the enterotoxin contaminant. PMID- 12733491 TI - Audit of scope and culture techniques applied to samples for the diagnosis of Mycobacterium bovis by hospital laboratories in England and Wales. AB - This audit examines the ability of English and Welsh laboratories to diagnose Mycobacterium bovis infection. All 164 clinical laboratories submitting samples to the PHLS Mycobacterium Reference Unit and Regional Centres for Mycobacteriology were surveyed. Twenty per cent of responding centres did not use a pyruvate-containing medium or incubate for the minimum recommended period of 8 weeks. This study demonstrates the potential for the underdiagnosis of M. bovis infection in England and Wales. Possible reasons for underdiagnosis are discussed together with strategies to optimize recovery of M. bovis. PMID- 12733492 TI - Miliary tuberculosis and acute respiratory distress syndrome. AB - OBJECTIVE: Miliary tuberculosis is a life-threatening disease caused by the haematogenous spread of Mycobacterium tuberculosis. We evaluated the clinical manifestations of 34 patients with miliary tuberculosis. DESIGN: A retrospective case review. RESULTS: The diagnosis of miliary tuberculosis was based on the identification of miliary nodules on chest radiography and one of the three following criteria: 1) acid-fast bacilli smear and/or culture positive in clinical specimens (22/34), 2) histopathological identification of TB granuloma (6/34), and 3) radiological and clinical improvement after anti-tuberculosis treatment (6/34). The median age (+/-SD) of the patients was 42.7 +/- 21.6 years, with two peaks, in the age group 20-30 and in those over 60. There were 16 underlying diseases in 14 patients, of which liver cirrhosis was the most common. The drug sensitivity pattern was available for 17 isolates of M. tuberculosis: 14 were sensitive, while the other three were resistant to at least one anti tuberculosis drug. Eight patients developed acute respiratory distress syndrome (ARDS), five of whom died during intensive care. Platelet count, serum albumin and liver enzyme level at the time of admission were significant factors both for ARDS development and for survival. CONCLUSION: ARDS caused by miliary TB is associated with a high fatality rate; scope remains for improvement in its management. PMID- 12733493 TI - [Acute adverse effects of dialysis]. AB - Adverse reactions to dialyzers are a not very frequent, but because of the serious, sometimes fatal course, a dreaded complication of haemodialysis treatment. Most important among these reactions are hypersensitive reactions (anaphylactoid, reaction type A to dialyzer), which develop as a rule within the 10th minute of the procedure, and the reaction caused by the action of perfluorohydrocarbon which develop hours after onset or even completion of haemodialysis. Explanation of the development of hypersensitive reactions (HSR) by complement activation and formation of anaphylatoxins C3a and C5a during contact of blood with the bioincompatible dialysis membrane has been abandoned. Evidence of the etiological role of ethylene oxide (ETO) in the development of HSR influenced the selection of materials for the production of dialyzers and sterilization during manufacture, it emphasized the importance of rinsing of the dialyzer in the dialysis centre and led to the wide application of alternative methods of sterilization by gamma radiation and steam. HSR may be also caused by overproduction of bradykinin and inhibition of its degradation or degradation of its metabolites. Excessive bradykinin production caused by dialysis membranes with a negative charge is potentiated e.g. by a lower pH and increased plasma dilution in the initial stage of haemodialysis. Inhibition of bradykinin degradation develops during treatment with angiotensin converting enzyme inhibitors (ACEI). In prevention of HSR associated with bradykinin in addition to elimination of a combination of a negatively charged dialysis membrane and ACEI treatment a part is played also by rinsing of the dialyzer before haemodialysis with a bicarbonate solution and the modification of the membrane surface (implemented by the manufacturer) which reduces its negative charge. The first reaction to the dialyzer in conjunction with perfluorohydrocarbon (PF-5070), used in production of some dialyzers for testing the integrity of their capillaries were first described in 2001 and have caused since then at least 50 deaths. Reactions associated with PF-5070 are still the subject of research and forensic investigations. Despite this they draw already now attention to the failure in manufacture of dialyzers, to the inadequacy of hitherto accepted testing procedures of dialyzers and the constant necessity of careful clinical, laboratory and post-mortem examination of dialyzed patients. PMID- 12733494 TI - [Bone changes in patients after partial gastric resection]. AB - Metabolic diseases of the bones may be a serious complication of gastrectomy. The pathogenesis of osseous changes is multifactorial. The most important causes include a reduced intake of dairy products, their fastened passage through the stomach and reduced calcium and vitamin D absorption. The objective of the work was to assess bone changes in patients after partial gastrectomy. From a total number of 261 patients with a minimal 10-year history of gastrectomy, in the investigation 31 patients were included (18 men and 13 women) who met the required criteria. The results were compared with a control group of 18 men and 13 women of equal age. The authors compared also groups of patients with the condition after surgery type Billroth I (8 patients) and Billroth II (23 patients). In all investigated subjects the authors examined the bone density in the area of the lumbar spine and neck of the femur and parameters of osteoresorption and osteoformation. The authors found a significant reduction of bone density in the area of the spine and neck of the femur (p < 0.001) and a significantly higher level of total alkaline phosphatase and intact parathormone in serum and N-terminal telopeptide collagen type I in urine (p < 0.001), as compared with the control group. The other parameters did not differ in the investigated groups. The authors did not find a statistically significant difference in the investigated parameters between patients after resection Billroth I and Billroth II. The results confirm the need of early diagnosis and treatment of bone changes in patients after partial gastrectomy. PMID- 12733495 TI - [Pitfalls in the diagnosis of carcinoids in patients treated with aminosalicylates]. AB - INTRODUCTION: Carcinoid is one of the most common endocrine active tumours of the gastrointestinal tract. 90% of all carcinoids originate from enterochromaffine cells in the GIT. In the literature the relationship of carcinoid of the bowel and IBD is mentioned, in particular Crohn's disease. The screening test used under our conditions is assessment of the excretion of the metabolite serotonin, 5-hydroxyindole acetic acid (HIAA) in urine. The authors wish to draw attention to falsely positive results of 5-HIAA in urine by the HPLC method in patients with CD treated with aminosalicylates (ASA). METHODS: In order to rule out carcinoid in chronically active CD the authors assessed after discontinuing known interfering drugs the excretion of HIAA by the HPLC method in 14 patients. The results were confirmed in laboratories of the Czech Academy of Sciences using mass spectrometry by desorption and ionization with a laser in the presence of matrix (MALDITOF MS), analytical procedures during processing of the specimens were modified according to Coward. In two patients urinary HIAA excretion was assessed on five consecutive days after discontinuation of ASA. RESULTS: The mean values of HIAA excretion by the HPLC method was highly suspicious of interference. Using the MALDI-TOF MS the authors did not detect 5-HIAA in the fraction of the interfering peak. After discontinuation of 5-ASA the interference disappeared after 4 days. By adjustment of the pH of the mobile buffer phase according to Coward the interfering peak was separated from the 5-HIAA peak. HIAA excretion assessed by the HPLC method was not significantly higher in patients after discontinuation of 5-ASA. CONCLUSION: The authors wish to draw attention to the possible development of carcinoid on the background of chronically active CD. Using assessment of urinary HIAA excretion by the HPLC method as a screening test it is essential to discontinue 5-ASA for at least 4 days before collection of urine or modify the analytical procedure when processing the specimen. PMID- 12733497 TI - [How to minimize the secondary effects of antiretrovirals]. PMID- 12733496 TI - [Clinical efficacy of using ximedon in surgical practice]. AB - Results of clinical trial of the new pyrimidine derivative--ximedone are presented. The drug was administered per os 0.5 g x 4 times a day (0.5 h before meals). Total course dose achieved 60 g. Ximedone implementation for complex treatment of burns, trophic ulcer of lower limbs, different inflammations demonstrated positive results. Prophylaxy administration of the drug in abdominal surgery at acute appendix resulted with lower incidence of postoperative complications. High efficacy and good tolerability of the drug allows to recommend it for wide medical use. PMID- 12733498 TI - [How antiretroviral medicines work]. PMID- 12733499 TI - [The importance of adherence to treatment]. PMID- 12733500 TI - [Bisegmental stability and Synex]. PMID- 12733501 TI - [Duties of the physician in the context of the current law on medical products]. PMID- 12733502 TI - [Concerning the article by Ischinger et al. "Reuse of "single use" medical devices after quality assured reprocessing: a model of cost savings?" Z Kardiol 91:889-898 (2002)]. PMID- 12733503 TI - tPA in daily clinical practice. PMID- 12733504 TI - Non-myeloablative allogeneic stem cell transplantation for metastatic renal cell carcinoma. PMID- 12733505 TI - [Persistent pulmonary inflammation in a patient who spent time in Asia. Melioidosis]. PMID- 12733506 TI - Female sex: a questionable risk factor for carotid endarterectomy. PMID- 12733508 TI - [Kaposi sarcoma-associated human herpesvirus 8 infection. With destabilization of cellular beta-catenin comes proliferation and tumor growth]. PMID- 12733507 TI - [Urological complications of endometriosis]. AB - Endometriosis (E) of the urinary tract is often not diagnosed at the beginning of the disease, particularly in cases with bladder wall involvement resulting in persistent dysuria and pelvic pain. Therefore, cystitis-like symptoms in younger women without evidence of urinary tract infections should be considered to be caused by E. Characteristic endoscopic findings may be missed and deep transurethral resection may be necessary for harvesting endometriotic tissue. This situation and improvement of diagnosis by ultrasound are demonstrated by a case report. The development of endometrial polyps in the uterus after tamoxifen (TAM) management is a well-known side effect of this antiestrogenic therapy. We observed a woman with endometrial polyps in the bladder after TAM. Endometriotic ureter stenosis in the absence of colics or other symptoms may results in irreversible loss of kidney function. Verification of the diagnosis is a common task of urology and gynecology. In three of six cases treated in our institution within 5 years, E of the ureter was first ascertained by the presence of ureter damage following surgical treatment of E. In two cases bowel E was present at the same time. Conservative treatment by suppression of ovarian function in most cases of stenosis of the ureter does not avoid the need for subsequent resection and reimplantation because of persistent fibrosis of the ureter wall. PMID- 12733509 TI - [Renal cell carcinoma in childhood. Case report and review]. AB - Renal cell carcinoma (RCC) in childhood is rare and its incidence is estimated to be 3-6% of all kidney cancers in childhood. Thus, clinical experience with renal cell carcinoma in this age group is meager and no consensus between pediatric urologists and pediatric oncologists exists on common treatment strategies. Furthermore, most previous reports included postpubertal patients in whom the biological behavior of the RCC and treatment options are the same as in adults. Based on the literature and a clinical case report of a 12-year-old boy suffering from a papillary renal cell carcinoma stage IV (pT2 N2 MO), the relevance of the biological behavior of different histological subtypes and their treatment options are reviewed and discussed. In adulthood lymph node dissection is still controversial, but in children it seems to have a positive effect on survival. Immunotherapy in childhood is still and experimental option and just a few cases are reported in the literature. PMID- 12733510 TI - [Diagnostics and therapy of diabetic nephrology]. AB - In Germany, 36% of all new chronic dialysis patients have diabetic nephropathy. The majority are type 2 diabetics. Early intervention has the greatest effect. Incipient nephropathy can be diagnosed by evidence of microalbuminuria (30-300 mg albumin/g creatinine). Proteinuria on the standard test strip (>300 mg/g) indicates manifest nephropathy followed by progressive renal failure. Important cofactors for progression are hypertension, hyperglycemia, and smoking. Low normal blood pressure levels (<130/80 mmHg without and <125/75 mmHG with proteinuria) based on ACE inhibitors/AT1 blockers are the goal. Combination therapies are frequently necessary. This can often reverse microalbuminuria. Chronic renal failure requires special attention (e.g. bone metabolism, anemia, acidosis). Timely initiation of renal replacement therapy (GFR <15 ml/min) reduces morbidity and mortality. In addition to hemo- and peritoneal dialysis, early kidney and in individual cases of type 1 diabetes combined kidney/pancreas transplantation is appropriate. PMID- 12733511 TI - Abstracts from the Environmental Mutagen Society 34th Annual Meeting. May 10-14, 2003. Miami Beach, Florida, USA. PMID- 12733512 TI - Abstracts of the British Society for Rheumatology XX Annual Meeting. 1-4 April 2003, Manchester, United Kingdom. PMID- 12733513 TI - Hyperglycemia and early reperfusion therapy. PMID- 12733515 TI - Abstracts of the American Geriatrics Society Annual Meeting. May 14-18, 2003. Baltimore, Maryland, USA. PMID- 12733514 TI - Abstracts of SCANNING 2003. San Diego, California, USA. PMID- 12733516 TI - Abstracts of the 38th Annual Meeting of the European Association for the Study of the Liver. March 29-April 1, 2003. Istanbul, Turkey. PMID- 12733517 TI - Abstracts of the Conference of the Romanian Society of Allergology and Clinical Immunology and the Romanian Society for Immunology. April 5-7, 2002. PMID- 12733518 TI - Active immunization against tumors. PMID- 12733519 TI - Abstracts of the International Conference on Cerebral Palsy. Quebec City, Canada, 30 April-3 May 2003. PMID- 12733520 TI - Orthopaedic proceedings: 2000, 2001, 2002. PMID- 12733521 TI - Long term care buyer's guide, 2000. PMID- 12733522 TI - Functional MRI: a potential physiologic indicator for stroke rehabilitation interventions. PMID- 12733523 TI - Funeral for a friend. PMID- 12733524 TI - Abstracts for the American Association of Clinical Endocrinologists 12th Annual Meeting and Clinical Congress. May 14-18, 2003. San Diego, California, USA. PMID- 12733525 TI - High-throughput structure determination. Proceedings of the 2002 CCP4 (Collaborative Computational Project in Macromolecular Crystallography) study weekend. January, 2002. York, United Kingdom. PMID- 12733527 TI - Author and subject index, volumes 51-75, 1998-2003. PMID- 12733526 TI - [Abstracts of the Neurology Meeting of the French Speaking Community. Nantes, France, 8-12 April 2003]. PMID- 12733528 TI - Abstracts of the Society for Pediatric Radiology 46th annual meeting. May 7-10, 2003, San Francisco, California, USA. PMID- 12733529 TI - [The hospital between values and interests: an historical perspective]. PMID- 12733531 TI - Infant formula supplemented with DHA: are there benefits? PMID- 12733530 TI - Grants to states for operation of qualified high risk pools. Final rule with comment period. AB - This final rule with comment period implements a provision of the Trade Assistance Reform Act of 2002 by providing $40 million in Federal fiscal year 2003 and $40 million in Federal fiscal year 2004 to States that have incurred losses in connection with the operation of qualified high risk pools that meet certain criteria. This grant program implements section 2745 of the Public Health Service Act, as added by the Trade Adjustment Assistance Reform Act of 2002. PMID- 12733532 TI - [XLVII Congress of the French National Society of Internal Medicine. Paris, France, 12-14 December 2002. Proceedings and abstracts]. PMID- 12733533 TI - Acrylamide: not so bad? PMID- 12733534 TI - Proceedings of the Conference on Healthy Cities and Health Promoting Universities and the Symposium on Health Education and Health Promotion. December 2001, Hong Kong. PMID- 12733536 TI - Current perspectives in fracture management and orthopaedic reconstruction. Proceedings of the RIVA 2000 Congress. PMID- 12733535 TI - Iron fulfillment possible through vegetarian lifestyle. PMID- 12733537 TI - Modern Periodontology--New Directions in the 21st Century. Symposium proceedings. Kanagawa, Japan. PMID- 12733538 TI - FDA clears home glycated hemoglobin test for diabetics. PMID- 12733539 TI - Researchers expand & preserve stem cells for cellular therapies. PMID- 12733541 TI - Setting the standard. PMID- 12733540 TI - [Diagnosis and treatment of pneumonia in adults and children (consensus of Lituanian pulmonologists)]. PMID- 12733543 TI - Evolutionary medicine puts symptoms in perspective. PMID- 12733542 TI - Thanks for professional support. PMID- 12733544 TI - Regarding the editorial on tobaccoism. PMID- 12733545 TI - What will it take to stop obesity? PMID- 12733547 TI - Research at US colleges of osteopathic medicine: a decade of growth. AB - Although research is a critical component of academic medicine, it has not been a significant component of osteopathic medicine. For years, leaders in the osteopathic medical profession have called for increased research in osteopathic medical schools. The need for cost-effective clinical practice leading to improved clinical outcomes creates a necessity for conducting well-designed clinical outcomes research related to osteopathic practice. The authors assess the growth in research at osteopathic academic medical centers from 1989-1999. The amounts of extramural funding at each school, sources of funding, types of research funded, departments funded, and investigators' degree types are also assessed. During the 10 years analyzed, total research funding increased 37%. Twenty-five percent of the grants and 55% of the funding to colleges of osteopathic medicine were from the National Institutes of Health. Most (63%) grants were awarded to PhD faculty. Most research was conducted in the basic biomedical sciences. Clinical research related to osteopathic practices appears to be a relatively minor component of research at osteopathic medical centers. PMID- 12733546 TI - An analysis of osteoporosis-related hip fractures using hospital discharge data. AB - The purpose of this study was to assess whether physicians in practice inadequately diagnose osteoporosis in a high-risk population of postmenopausal women who have sustained hip fractures. Using the Texas Hospital Discharge Data Public Use Data File (PUDF) provided through the Texas Health Care Information Council, the authors conducted a review of all postmenopausal women older than 55 years with fractured hips discharged from Texas hospitals during 1999. A total of 13,628 patients meeting these criteria were found using the PUDF. In their diagnoses, physicians for 2233 (16.3%) of these 13,628 women also specified the code for osteoporosis (P < .001) from the ninth revision of the International Classification of Diseases. It is estimated that between 40% and 50% of postmenopausal women have osteoporosis. Therefore, women with fragility fractures form an even more at-risk subset of the population--so much so that one would expect a majority of these women to carry diagnoses of osteoporosis. The age distribution in each group was comparable, implying that receiving a coded diagnosis for osteoporosis was not related to the age of the patient when she was admitted to the hospital. Further, when data was analyzed by race or ethnicity, percentages for each group (ie, diagnosed with hip fracture only versus diagnosed with hip fracture and osteoporosis) were comparable. In conclusion, physicians practicing in Texas during calendar year 1999 inadequately diagnosed osteoporosis in a high-risk population of postmenopausal women who were admitted to hospitals with fractured hips. Future analysis of subsequent annual databases will identify whether continuing medical education efforts cause physicians to diagnosis osteoporosis in this high-risk population more frequently. PMID- 12733548 TI - An adaptable, transportable web-based data acquisition platform for clinical and survey-based research. AB - Given the need for multicenter clinical trials to support evidence-based treatments, the authors hypothesized that the process of gathering large amounts of data from disparate clinical sites could be facilitated through direct input of clinical and survey data through a Web interface. A series of data collection instruments was created and published as Web pages to support a clinical study performed at Des Moines University Osteopathic Medical Center. The most challenging tool to implement was the visual analog scale, which required special programming. Specific Web pages allowed research study participants to input their own data, while other Web pages were restricted to use by the investigator for inputting clinical and laboratory observations. Data from these sources were automatically combined in a single spreadsheet by the Web administrator in a manner that maintained the confidentiality of participants. Subsequently, the system was tested from a remote site (Chicago), and data were captured at Des Moines University Osteopathic Medical Center. This test involved a variety of data including the visual analog scale. Although this system provided a facile method for collecting and analyzing a large amount of data in almost real time, with a demonstrable savings of time and money, the authors believe the more important use for this data collection system is in large multicenter clinical trials. Hence, the authors commend its use for support of large outcomes studies for osteopathic researchers engaged in interinstitutional efficacy studies. PMID- 12733549 TI - A myoelectric model for thoracic spinal motion dynamics during clinical rotation tests: Part 1. Ipsilateral regional motor performance. AB - Osteopathic physicians may use regional diagnostic rotation tests of the spine during physical examinations for patient evaluations. Clinical judgments of these responses relate to symmetry as a criterion for mobility. This first part of a two-part study reports the authors' investigation of regional ipsilateral myoelectric activity during responses to active and passive shoulder and trunk rotations on the left- and right-side muscles with subjects seated. Results indicate symmetry existed in both active and passively induced regional rotation tests. Further, the distribution (profile) of ipsilateral myoelectric activity in the thorax reflected a bell-shaped activity curve that peaked at thoracic levels 6 and 7. This profile element demonstrates remarkable similarity between volitional and physician-induced rotation motions. These myoelectric data justify symmetry and profile as tangible standards for making clinical judgments of regional responses to spinal rotation diagnostic tests. PMID- 12733550 TI - Measuring the performance of screening mammography in community practice with Medicare claims data. AB - BACKGROUND: Few studies have examined the outcomes of screening mammography in community practice, particularly the extent of false positive exams among older asymptomatic women. RESEARCH DESIGN: Subjects were female Medicare beneficiaries, age 67 or older, residing in one of eleven SEER areas, with no evidence of breast cancer. Medicare claims data were used to identify their screening mammograms over two time periods, 1993-1995 and 1996-1998, and to measure their use of follow-up diagnostic testing (diagnostic mammography, breast ultrasound and breast biopsy) within three months of the screening mammogram. RESULTS: There were significant differences among the rates of diagnostic testing for each age group (67-74; 75+ ) by year, but no clear trend toward higher or lower rates over time. Although rates of diagnostic testing differed significantly by geographic region in both time periods 1993-1995 and 1996-1998, estimates of specificity for all regions were within AHRQ clinical practice guidelines (specificity greater than 90%). Specificity significantly improved with the volume of the radiologist's practice for the latter time period (1996-1998) but not for the former (1993-1995). CONCLUSION: Medicare claims offer an accessible population based source of data for mammography performance indicators. As such, they offer a low cost method for evaluating individual mammography practices as well as monitoring the impact of reimbursement policies, practice guidelines and laws mandating requirements for accrediting facilities. PMID- 12733551 TI - Paid and unpaid work, and its relation to low back and neck/shoulder disorders among women. AB - The aim of the present study was to estimate the influence of total work hours, paid work in the labor market and unpaid work in the family domain, on care seeking for low back and neck/shoulder disorders in the female population. The exposure assessments considered a typical working' day during the previous twelve months and were assessed by interviews and questionnaires; 704 cases and 984 referents were examined. The cases had sought professional care during the study period; the referents were randomly selected from the same source population. There was no increased relative risk for care-seeking for low back and neck/shoulder disorders for gainfully employed women compared to those not employed, or for full-time compared to part-time working women. At least 60 hours per week of paid work, or at least 40 hours per week of unpaid work, separately, indicated an increased relative risk for care-seeking. The present results did not strengthen the hypothesis that a high amount of hours of work is an independent risk factor for musculoskeletal disorders. PMID- 12733552 TI - From canteen to lunch box: ergonomic demands in distribution of portion-packed hot food. AB - The present case-study, which is rather small in number of subjects but has a broad perspective, is part of a larger investigation designed to initiate development processes in working life in one specific region in Sweden. This study may serve as an example of ergonomic fieldwork with a gender perspective. AIM: The overall aim was to examine the physical and psychosocial working conditions among a group of female hot food distributors, and to relate these conditions to other traditionally heavy work within the same working unit, as well as to suggest improvements. METHOD: The study group consisted of ten female municipal employees in a provincial town. Structured interviews about the psychosocial working conditions, measurements of heart rate, ratings of self reported perceived exertion and observations of one working day were performed. RESULTS: The overall impression from the study was that the food distributors in several aspects had a very difficult working situation that ought to be improved. The food distributors reported high psychological demands and low decision latitude, time pressure and dissatisfaction with not being able to live up to the pensioners' and the employer's expectations. They also had a high heart rate increase during work, which may be a risk factor for impaired health. Several short-term and long-term solutions were suggested to reduce and redistribute the total workload. DISCUSSION: The employer has attended to some suggested short term solutions and the follow-up study showed that the working conditions had been improved. A reduction of cardiovascular load, as well as self-rated physical exertion, was noted and the work was perceived as substantially less stressful. PMID- 12733553 TI - A comparison of the mental health of employed and unemployed women in the context of a massive layoff. AB - The purpose of this study was to assess the impact of unemployment on the mental health of women in the context of massive unemployment. Comparisons were made between the level of mental distress experienced by unemployed and employed women, in two areas of Newfoundland, Canada that were affected by the northern cod moratorium. In addition, the relationships between women's mental distress and a number of variables were explored. Questionnaires were administered to 112 unemployed and 112 employed women three years after the moratorium began. The unemployed women reported significantly poorer mental well-being in the year prior to data collection. At the time of the study, however, both groups of women were experiencing high levels of distress. The moratorium, financial problems, and feelings of uncertainty were identified as key stressors for all the women, but especially for those without work. Among the working women, past experience with unemployment and level of education had significant correlations with their mental well-being. PMID- 12733554 TI - Abortion beliefs and practices among midwives (parteras) in a rural Mexican township. AB - Reproduction and motherhood are among the most important components of women's identity throughout Mexico and, for many women, are the only vehicles for gaining recognition and status in the family and community. At the same time, however, abortion is a central experience in the lives of many women and carries with it the complexities and contradictions of women's reproductive and sexual health. This paper presents results from an ethnographic study conducted with midwives in one rural township of Morelos, Mexico to understand their conceptualizations of and practices related to abortion and postabortion care. Overall, midwives viewed miscarriage as a woman's failure to fulfill her primary role as mother and induced abortion as a grave sin or crime. Nevertheless, under certain circumstances induced abortion was justified for many midwives. Helping women to "let down the period" in situations when a woman's menstrual period was delayed was acceptable to midwives as it was not viewed as abortion and enabled women to regain health and well-being. PMID- 12733555 TI - Why do women decline prenatal screening and diagnosis? Australian women's perspective. AB - In this paper, we examine reasons for declining both prenatal screening, and diagnosis among a small group of pregnant women in Victoria, Australia. Semi structured questionnaires were used to elicit women's account of their refusals of offers during pregnancy. Previous literature suggests that women decline prenatal screening and diagnosis because they are against abortion and the medicalisation of pregnancy, and have conoerns about the health and well-being of their fetuses. Women in this study had similar reasons but they also had other reasons for declining. Most clearly was that related to 'risk' brought about by the prospect of knowledge gained from undertaking prenatal screening and diagnostic tests, which would cause emotional distress and lead to further difficult decisions. The results have implications for the development and expansion of prenatal screening and diagnosis for pregnant women in Australia and elsewhere. We are not suggesting that prenatal screening and diagnosis is a problem for all women or even most women. However, health service providers must provide information about prenatal screening and diagnosis that is appropriate for all pregnant women, presenting all options, including that of not having any screening or diagnostic test. In doing so we will be facilitating the opportunity for women to make an informed choice and acknowledging the existence and importance of this small, but concerned group of women. PMID- 12733556 TI - Moves to eliminate 'anticompetitive' practices in the supply of prescription-only veterinary medicines. PMID- 12733557 TI - Clinical and laboratory investigations of five outbreaks of foot-and-mouth disease during the 2001 epidemic in the United Kingdom. AB - Clinical and laboratory investigations of five outbreaks of foot-and-mouth disease (FMD) were made during the early stages of the 2001 epidemic in the UK. The first outbreak, confirmed on February 20, was at an abattoir in Essex which specialised in the processing of culled sows and boars. On February 23, the disease was confirmed at a pig farm in Northumberland which held cull sows and boars fed on waste food; the findings indicated that it was the first of the five premises to be infected. The disease had probably been present since early February, and it was the most likely origin of the epidemic. The other premises investigated were a waste food-fed cull sow/boar pig unit in Essex, approximately 30 km from the abattoir, which was probably infected at the same time or before the abattoir, a sheep and cattle farm approximately 6 km from the Northumberland pig farm, which was probably infected by airborne virus from it in the period immediately before February 13, and a sheep and cattle farm in Devon which had clinical disease from February 20 and was probably infected by sheep transported from Northumberland on February 13 which arrived on February 15. PMID- 12733558 TI - Survey of permanent wound tracts in the carcases of culled wild red deer in Scotland. AB - The number and sites of permanent wound tracts in the carcases of 943 wild culled red deer (Cervus elaphus) were recorded. During the peak period of the red deer rut there was a significant increase in the number of these tracts in the carcases, which was associated with a decrease from 89 per cent to 71 per cent in the probability of the first permanent wound tract also being the last (the terminal probability). There were significantly more permanent wound tracts in the carcases of one group (predominantly males) than in a second group (predominantly females and calves). In carcases with a single tract in the trunk, in which the heart and lungs were also examined, 80 per cent had tracts involving the heart and/or lungs, the recommended thoracic target organs. Tracts involving vital structures in the neck were also common, with 15.3 per cent of the carcases with a single permanent tract having damage limited to cervical structures. PMID- 12733559 TI - Incidence of cryptorchidism in dogs and cats. AB - Over a period of 54 months, 3518 dogs and 3806 cats were castrated; 240 of the dogs and 50 of the cats were cryptorchid. Pedigree dogs, in particular the German shepherd dog, boxer and chihuahua were over-represented. Among the dogs, right sided inguinal cryptorchidism was the most common form, followed by right-sided abdominal cryptorchidism. The location of the affected testicle(s) was most variable in the boxer. Among the cats, left- or right-sided inguinal cryptorchidism were the most common forms of the condition. PMID- 12733560 TI - Exploratory findings on the prevalence of contagious ovine digital dermatitis in sheep in England and Wales during 1999 to 2000. PMID- 12733561 TI - Thiazole orange-positive platelets in cats with thrombocytopenia induced by cyclophosphamide. PMID- 12733562 TI - Nodular hyperplasia and cysts in thyroid glands of llamas (Lama glama) from north west USA. PMID- 12733563 TI - Veterinary surveillance in the UK. PMID- 12733564 TI - Kidney transplants: background to the ethical debate. PMID- 12733565 TI - Footrot in sheep. PMID- 12733566 TI - Control of FMD by vaccination. PMID- 12733567 TI - Palmate newt mortality at an English breeding site. PMID- 12733568 TI - Characterization of principal nutritional components of Brazilian oil palm (Eliaes guineensis) fruits. AB - In spite of the fact that most of the members of Palmaceae contain high concentrations of oil, its potential as a source of oil and protein for human consumption has not been exploited. The pulp and kernels of the Eliaes guineensis palm fruits grown in the Northeast region of Brazil were analyzed only for their proximate composition. The lipid content of the dried pulp and kernels was 73.2% and 32.6%, respectively. Hexane extracted oils from the pulp and kernels yielded similar refractive indices, specific gravity but different peroxide, acid, iodine and saponification values. Gas chromatographic analysis revealed the presence of 24 and 18 fatty acids in pulp and kernel oils, respectively. The principal saturated acid of the pulp oil was palmitic acid (36.9% of the total), and lauric acid (53.3%) for kernel oil. Oleic acid was the predominant monounsaturated fatty acid in both the oils though its concentration in the pulp and kernel oils was 45.29% and 5.5%, respectively. In relation to the essential amino acids, pulp proteins presented a better profile than the kernel proteins. In comparison to the FAO reference protein, the pulp proteins were deficient in methionine, lysine and threonine (16.8%, 51.6% and 93.5% of FAO reference protein) but contained leucine, valine, isoleucine and phenylalanine in optimal concentrations. With exception to phenylalanine and valine (102.2% and 111.4% of reference protein, respectively), the kernel proteins were deficient in all other essential amino acids. The oils from this palm can be used as culinary oil and in margarine manufacture, while pulp could be a supplement for essential amino acids leucine, isoleucine, phenylalanine and valine with other protein sources that are deficient in these amino acids. PMID- 12733569 TI - Initial crop growth in soil collected from a closed animal waste lagoon. AB - In the 21st century, remediation of the soil beneath animal waste lagoons will become an important issue, as they are closed due to environmental regulations or to abandonment. The possibility of growing crops in the soil, which has high concentrations of ammonium-N, has not been studied. The objective of this experiment was to determine if crop species would germinate and grow in lagoon soil. Soil was gathered from a lagoon that had received wastes from swine (Sus scrofa) and beef (Bos taurus) since 1968. Eight crops were grown in greenhouse pots containing the lagoon soil: winter barley (Hordeum vulgare L. 'Weskan'); field corn (Zea mays L., Cargill's hybrid 7997); 'Plainsman' winter rapeseed [Brassica napus L. spp. oleifera (Metzg.) Sinsk. f. biennis]; soybean [Glycine max (L.) Merr. 'KS 4694'); forage sorghum [Sorghum bicolor (L.) Moench 'Norkan']; sunflower (Helianthus annuus L. 'Hysun 354'); and winter wheat (Triticum aestivum L.)--two cultivars: '2137' and 'Turkey.' Plants were grown for 35 days in lagoon soil or an agricultural soil (Haynie very fine sandy loam; coarse-silty, mixed, superactive, calcareous, mesic Mollic Udifluvent) obtained from a field near the closed lagoon. Ammonium-N (average value of 692 mg/kg) was about 70-85 times greater than the average value of 8-10 mg/kg NH4-N in Kansan soils. The lagoon soil was nonsodic and had a salinity ranking of "medium" with an electrical conductivity averaging 2.29 dS/m. The high ammonium-N concentration in the lagoon soil was not inhibitory to emergence and growth. The eight crops grew taller in the lagoon soil than in the agricultural soil. Except for '2137' wheat, dry weight was higher in the lagoon soil than in the agricultural soil. The results showed that the lagoon soil is not detrimental to early growth of eight crops. PMID- 12733570 TI - Studies on chromium(VI) adsorption-desorption using immobilized fungal biomass. AB - The aim of this study was to investigate the Cr(VI) biosorption potential of immobilized Rhizopus nigricans and to screen a variety of non-toxic desorbing agents, in order to find out possible application in multiple sorption-desorption cycles. The biomass was immobilized by various mechanisms and evaluated for removal of Cr(VI) from aqueous solution, mechanical stability to desorbents, and reuse in successive cycles. The finely powdered biomass, entrapped in five different polymeric matrices viz. calcium alginate, polyvinyl alcohol (PVA), polyacrylamide, polyisoprene, and polysulfone was compared for biosorption efficiency and stability to desorbents. Physical immobilization to polyurethane foam and coir fiber was less efficient than polymer entrapment methods. Of the different combinations (%, w/v) of biomass dose compared for each matrix, 8% (calcium alginate), 6% (polyacrylamide and PVA), 12% (polyisoprene), and 10% (polysulfone) were found to be the optimum. The Cr sorption capacity (mg Cr/g sorbent) of all immobilized biomass was lesser than the native, powdered biomass. The Cr sorption capacity decreased in the order of free biomass (119.2) > polysulfone entrapped (101.5) > polyisoprene immobilized (98.76) > PVA immobilized (96.69) > calcium alginate entrapped (84.29) > polyacrylamide (45.56), at 500 mg/l concentration of Cr(VI). The degree of mechanical stability and chemical resistance of the immobilized systems were in the order of polysulfone > polyisoprene > PVA > polyacrylamide > calcium alginate. The bound Cr(VI) could be eluted successfully using 0.01 N NaOH, NaHCO3, and Na2CO3. The adsorption data for the native and the immobilized biomass was evaluated by the Freundlich isotherm model. The successive sorption-desorption studies employing polysulfone entrapped biomass indicated that the biomass beads could be regenerated and reused in more than 25 cycles and the regeneration efficiency was 75-78%. PMID- 12733571 TI - Biosorption of Pb(II) and Cu(II) by activated sludge in batch and continuous-flow stirred reactors. AB - Biosorption of Pb(II) and Cu(II) ions in single component and binary systems was studied using activated sludge in batch and continuous-flow stirred reactors. In biosorption experiments, the activated sludge in three different phases of the growth period was used: growing cells; resting cells; dead or dried cells. Because of the low adsorption capacity of the non-viable activated sludge especially in the case of Pb(II) ions, biosorption of the Cu(II) and Pb(II) ions from the binary mixtures was carried out by using the resting cells. The biosorption data fitted better with the Freundlich adsorption isotherm model. Using a mathematical model based on continuous system mass balance for the liquid phase and batch system mass balance for the solid phase, the forward rate constants for biosorption of Pb(II) and Cu(II) ions were 0.793 and 0.242 1 (mmolmin)(-1), respectively. PMID- 12733572 TI - Preparation of polyol esters based on vegetable and animal fats. AB - The possibility of using some natural fats: rapeseed oil, olive oil and lard, as starting material for the preparation of neopentyl glycol (NPG) and trimethylol propane (TMP) esters is reported. The syntheses of final products were performed by alcoholysis of fatty acid methyl esters, obtained from natural fats studied, with the appropriate polyhydric alcohol using calcium methoxide as a catalyst. The basic physicochemical properties of the NPG and TMP esters synthesized were the following: viscosity at 40 degrees C in the range of 13.5-37.6 cSt, pour point between -10.5 and -17.5 degrees C and very high viscosity indices, higher than 200. Generally, the esters of neopentyl alcohols were characterized by higher stability in thermo-oxidative conditions in comparison to native triglycerides. Due to the low content of polyunsaturated acids, the olive oil based esters showed the highest thermo-oxidative resistance. Also, methyl esters of fatty acids of lard would constitute a good raw material for the synthesis of lubricating oils, provided that their saturated acids content was lowered. This permits synthesis of NPG and TMP esters with a lower pour point (below -10 degrees C) than natural lard (+33 degrees C). PMID- 12733573 TI - Phenolic compounds and antioxidant activity from red grape marc extracts. AB - The aims of this work were to determine the amounts of the different classes of phenolic compounds in an ethanolic extract from red grape marc and its components, peels and seeds, and to evaluate their antioxidant activities by the beta-carotene bleaching test for their utilization as natural antioxidants. The results showed that red grape marc was rich in polyphenol compounds with a clear antioxidant activity. The extracts, in fact, at very low concentration (20 ppm) in total phenols showed an antioxidant activity (AA) higher than 43% on average, while at higher concentration (80-160 ppm) all the fractions had an AA comparable to that of butlylated-hydroxytoluene. Grape seeds seemed to give the highest contribution to such AA, as they contained high quantities of proanthocyanidines, a type of flavonoid known for its high antioxidant properties. PMID- 12733575 TI - Comparative evaluation of yeast and bacterial treatment of high salinity wastewater based on biokinetic coefficients. AB - This paper compares the performance of the aerobic treatment of high organic-high salinity wastewater by yeast and bacterial systems. The biokinetic coefficients for both the systems were determined and used to analyze the behavior of the yeast and bacterial systems under high salinity conditions. It was found that the yeast culture was more efficient compared to the bacterial culture, especially for high salinity conditions that severely inhibit growth and performance of bacterial systems. The values of the biokinetic coefficients obtained from this study are in agreement with the observations. Nutrient removal capacity has also been found to be better for yeast due to higher nutrient uptake in the yeast biomass. PMID- 12733574 TI - Use of response surface methodology for selection of nutrient levels for culturing Paecilomyces variotii in eucalyptus hemicellulosic hydrolyzate. AB - Eucalyptus hemicellulose was hydrolyzed by treating eucalyptus wood chips with sulfuric acid. The hydrolyzate was used as the substrate to produce single-cell protein by growing Paecilomyces variotii IOC-3764 for 72 or 96 h. The influences of rice bran, ammonium sulfate and fermentation time were verified by a 23 full factorial central composite design. At the optimum process conditions, the cell concentration was 12.06 g/l, which was obtained when the microorganisms were cultivated for 89 h in a medium composed of 10 g/l rice bran, 2.0 g/l nitrogen and 1.1 g/l sodium phosphate. The mathematical model Y = 10.65 + 2.40X2 + 2.36X3 + 1.16X2X3 - 2.10X2(2) - 1.06X3(2) describes biomass production by P. variotii in eucalyptus hemicellulosic hydrolyzate with a determination coefficient of R2 = 0.9561, where X2 and X3 are ammonium sulfate and fermentation time, respectively. PMID- 12733576 TI - Production of alpha amylase by Bacillus licheniformis using an economical medium. AB - The present study is concerned with the selection of new medium for the production of alpha amylase by Bacillus licheniformis. Different agricultural by products such as wheat bran, sunflower meal, cotton seed meal, soybean meal, rice husk or rice bran were tested for the production of alpha amylase. Among different agricultural by-products evaluated, wheat bran was found to be the best basal and standardized medium for optimal production of alpha amylase. The production was increased 2-folds when soluble starch was replaced with pearl millet starch at 1% level and nutrient broth concentrations was reduced from 1% level to 0.5%. The newly selected fermentation medium containing (% w/v) wheat bran 1.25, nutrient broth 0.5, pearl millet starch 1.0, lactose 0.5, NaCl 0.5, CaCl2 0.2 in 100 ml of phosphate buffer. The kinetic values of Y(p/x), Y(p/s), and Q(p) indicated that the production of enzyme was greater in newly selected medium than the conventional more expensive medium. PMID- 12733577 TI - Microwave esterification of sunflower proteins in solvent-free conditions. AB - The esterification of storage proteins from sunflower seeds renders these molecules more hydrophobic and increases internal plasticization, facilitating their use in the fabrication of materials by thermo-mechanical processes. The reaction was carried out in solvent-free conditions to simplify the process and to reduce costs. Optimization of esterification by the classical method, heating in a thermostat-regulated oil bath, resulted in 84% esterification for a reaction time of 4 h (amount of catalyst = 3.9 meq 5 N HCl/g protein, T = 90 degrees C). Microwave heating was then used to reduce the reaction time and the hydrolysis of protein chains. A similar level of esterification (89%) was obtained in 18 min (amount of catalyst = 3.4 meq 5 N HCl/g protein, microwave power (P) = 560 W). High yields were obtained with this method, demonstrating that this process limited hydrolysis. PMID- 12733578 TI - Roles of silica gel in polycondensation of lactic acid in organic solvent. AB - Poly(lactic acid) is among the most important biodegradable, biocompatible polymers. To explore the feasibility of making poly(lactic acid) through potentially more selective enzymatic methods, the lipase-catalyzed direct polycondensation of lactic acid in organic solvents was investigated. At 37 degrees C the reaction was found to favor nonpolar solvents with larger log P values and smaller log S(w/o values. The addition of silica gel appeared to greatly enhance the lactic acid conversion (up to 98%) and the lipase stability under the reaction condition. However, upon further investigations, the silica gel itself was found to catalyze the polycondensation, in addition to the role of water removal. The conversion catalyzed by silica gel alone was actually higher than that by silica gel + lipase (or lipase alone). Up to 93% conversion of the acid functional group (or about 99.5% conversion of lactic acid monomer) was obtained in 120 h with silica gel as the catalyst. The finding is especially significant for interpreting (or reconsidering) the results of many presumably enzyme-catalyzed organic-phase reactions in the presence of silica gel. PMID- 12733580 TI - Biodegradation of hexadecane in liquid and solid-state fermentations by Aspergillus niger. AB - The biodegradation and mineralisation of hexadecane (HXD) by Aspergillus niger were studied in SmF and Solid-state fermentation (SSF). HXD concentrations ranging from 45 to 180 g/l (SSF) and from 20 to 80 g/l (SmF) were tested. HXD consumption was three times higher and fungal growth was up to 30 times faster in SSF than in SmF. The maximum HXD consumption in SmF was 62% (18% mineralised) and in SSF 100% (52% mineralised) for initial HXD concentrations of 20 and 45 g/l, respectively. The respiratory quotient in SmF increased (from 0.85 to 1.08) with increase in HXD concentration, while it was independent (approximately 0.68) of the initial HXD concentration in SSF. These results showed that the consumption rate and biodegradation efficiency for HXD were higher in SSF than in SmF. PMID- 12733579 TI - Biological denitrification in a continuous-flow pilot bioreactor containing immobilized Pseudomonas butanovora cells. AB - Pseudomonas butanovora, a novel denitrifying bacterium, was immobilized in composite beads and filled into a reactor system. The pilot bioreactor average denitrification activity was at ethanol-C:nitrate-N ratios of 3:1 and 1.5:1 0.88 and 0.54 kg NO3(-)-Nm(-3) d(-1), respectively. The denitrification was stable in spite of the relatively low hydraulic retention times of 2.47 and 3 h. The nitrate content of the influent was almost completely reduced at the first level of the bioreactor and the nitrite formed underwent reduction in the upper part of the reactor. The experimentally determined optimum ethanol-C:nitrate-N ratio was 1.41 +/- 0.41. In consequence of the aerobic conditions, the acetic acid produced by the oxygenation of ethanol was also detectable in the reactor effluent. The pH of the effluent (7.58) never exceeded the acceptable maximum (8.5). The nitrate removal efficiency of the cells was nearly 1000% at both C:N ratios, and the nitrite content of the effluent was around the prescribed limit throughout the continuous operation. This continuous-flow pilot bioreactor containing immobilized P. butanovora cells proved an efficient denitrification system with a relatively low retention time. PMID- 12733581 TI - Study on anaerobic and aerobic degradation of different non-ionic surfactants. AB - Six alcohol ethoxylates (C5E2, C6E4, C7E4, C8E2, C8E4, C10E4) and two fatty acid esters were tested at lab-scale for degradation in anaerobic and aerobic conditions and oxygen uptake rate (OUR). Anaerobic removal of C5E2, C6E4 and C7E4 improved with increasing number of ethoxy groups (E) and decreasing length of the alkyl chain (C). Their aerobic removal was also great but lower than the anaerobic values. C8E2, C8E4 and C10E4 were adsorbed on sludge but not degraded in anaerobic conditions, while they were efficiently removed under aerobiosis. The fatty acid esters were removed to a level between the two alcohol ethoxylates groups in both anaerobiosis and aerobiosis. The measured OUR confirmed the different behaviours of the three groups of compounds. PMID- 12733582 TI - Dynamics of carbon, nitrogen and phosphorus in soil amended with irradiated, pasteurized and limed biosolids. AB - Sewage biosolids contain high concentrations of pathogens, which limits their use as soil amendment. This study investigated how application of lime (Ca(OH)2), irradiation, or pasteurization reduced pathogens in biosolids and how its application affected soil characteristics. A soil sampled outside the canopy of Mesquite trees (Prosopis laevigata) and from a pasture at Lerma (Mexico) was amended with treated or untreated biosolids, characterized and incubated aerobically while dynamics of carbon (C), nitrogen (N) and phosphorus (P) were monitored. Heavy metals concentrations in the biosolids were low, so it was of excellent quality (USEPA). The amount of pathogens in the biosolids made it a class "B" (USEPA) which can be used in forests. Only irradiation sufficiently reduced faecal coliforms to make it a class "A" biosolids without restrictions in application. C mineralization increased significantly when biosolids were added, but not concentrations of available P (P < 0.05). Ammonium (NH4+) concentrations in soil amended with biosolids were higher compared to unamended soil, but not the concentrations of nitrate (NO3-) except when biosolids treated with Ca(OH)2 was added to the Lerma soil. PMID- 12733583 TI - Nitrification, denitrification and biological phosphorus removal in piggery wastewater using a sequencing batch reactor. AB - Nutrients in piggery wastewater with high organic matter, nitrogen (N) and phosphorus (P) content were biologically removed in a sequencing batch reactor (SBR) with anaerobic, aerobic and anoxic stages. The SBR was operated with 3 cycles/day, temperature 30 degrees C, sludge retention time (SRT) 1 day and hydraulic retention time (HRT) 11 days. With a wastewater containing 1500 mg/l ammonium and 144 mg/l phosphate, a removal efficiency of 99.7% for nitrogen and 97.3% for phosphate was obtained. Experiments set up to evaluate the effect of temperature on the process showed that it should be run at temperatures higher than 16 degrees C to obtain good removals (> 95%). Batch tests (ammonia utilization rate, nitrogen utilization rate and oxygen utilization rate) proved to be good tools to evaluate heterotrophic and autotrophic biomass activity. The SBR proved to be a very flexible tool, and was particularly suitable for the treatment of piggery wastewater, characterized by high nutrient content and by frequent changes in composition and therefore affecting process conditions. PMID- 12733584 TI - Mathematical modeling of non-ideal mixing continuous flow reactors for anaerobic digestion of cattle manure. AB - Most conventional digesters used for animal wastewater treatment include continuously stirred-tank reactors. While imperfect mixing patterns are more common than ideal ones in real reactors, anaerobic digestion models often assume complete mixing conditions. Therefore, their applicability appears to be limited. In this study, a mathematical model for anaerobic digestion of cattle manure was developed to describe the dynamic behavior of non-ideal mixing continuous flow reactors. The microbial kinetic model includes an enzymatic hydrolysis step and four microbial growth steps, together with the effects of substrate inhibition, pH and thermodynamic considerations. The biokinetic expressions were linked to a simple two-region liquid mixing model, which considered the reactor volume in two separate sections, the flow-through and the retention regions. Deviations from an ideal completely mixed regime were represented by changing the relative volume of the flow-through region (a) and the ratio of the internal exchange flow rate to the feed flow rate (b). The effects of the hydraulic retention time, the composition of feed, the initial conditions of the reactor and the degree of mixing on process performance can be evaluated by the dynamic model. The simulation results under different conditions showed that deviations from the ideal mixing regime decreased the methane yield and resulted in a reduced performance of the anaerobic reactors. The evaluation of the impact of the characteristic mixing parameters (a) and (b) on the anaerobic digestion of cattle manure showed that both liquid mixing parameters had significant effects on reactor performance. PMID- 12733585 TI - Quality assessment of compost prepared from fly ash and crop residue. AB - Fly ash was co-composted with wheat straw and 2% rock phosphate (w/w) for 90 days and different chemical and microbiological parameters monitored to evaluate its effect on the composting process. Fly ash addition at 20% level resulted in the lowest C/N of 16.4:1 and highest available and total phosphorus. Increasing the addition of fly ash from 40 to 60% (w/w) did not exert any detrimental effect on either C:N or the microbial population. PMID- 12733586 TI - Utilization of various agricultural wastes for activated carbon preparation and application for the removal of dyes and metal ions from aqueous solutions. AB - Activated carbons were prepared from the agricultural solid wastes, silk cotton hull, coconut tree sawdust, sago waste, maize cob and banana pith and used to eliminate heavy metals and dyes from aqueous solution. Adsorption of all dyes and metal ions required a very short time and gave quantitative removal. Experimental results show all carbons were effective for the removal of pollutants from water. Since all agricultural solid wastes used in this investigation are freely, abundantly and locally available, the resulting carbons are expected to be economically viable for wastewater treatment. PMID- 12733587 TI - Calotropis procera (Sodom apple)--a potential material for enzyme purification. AB - A simple method based on precipitation with Calotropis procera latex was developed for the purification of crude enzyme from fermentation broth. C. procera latex (10(-2) dilution) clarified and concentrated the crude amylase of Aspergillus oryzae 4-fold with 97% recovery of the initial amylase activity in the filtrate in a single step operation. The latex was stable at pH < or = 4.5 and there was no significant difference (P < or = 0.05) in the purification potential of the latex at 4 and 28 degrees C. This method could prove useful to developing economies of the tropics where ambient temperature is around 28 degrees C. PMID- 12733589 TI - Distinct T cell subsets and cytokine production in cultures derived from transformation zone and squamous intraepithelial lesion biopsies of the uterine cervix. AB - PROBLEM: The characterization of lymphocytes issued from squamous intraepithelial lesions (SIL) and from the transformation zone (TZ), where the majority of SIL occur, is important to understand the role of immunity in SIL development. METHOD OF STUDY: We compared lymphocyte populations of the TZ and SIL with those of normal exocervix, using a technique allowing for the isolation of lymphocytes, either from the epithelium or from the underlying stroma of small biopsies. RESULTS: The majority of cells derived from the epithelium of all biopsies were CD8+ T cells. Some SIL-derived cultures were characterized by an increased proportion of activated TCRgammadelta+. The production of the immunosuppressive cytokine IL10 was significantly higher in lymphocyte cultures from the normal TZ in comparison with the exocervix. A decreased percentage of effector T cells was observed in cultures derived from the stroma of normal TZ (TCRgammadelta+) or SIL (CD8+) in comparison with the exocervix. CONCLUSIONS: Our results suggest that a low proportion of effector T cells and IL10 production could contribute to the predisposition of the TZ to the development of SIL and to the progression of SIL to cervical cancer. PMID- 12733588 TI - Progesterone regulates IL12 expression in pregnancy lymphocytes by inhibiting phospholipase A2. AB - PROBLEM: Progesterone-induced blocking factor (PIBF) is one of the pathways that mediate the immunological effects of progesterone. PIBF inhibits natural killer (NK) cytotoxic activity. Recently we showed that neutralization of PIBF results in an increased interleukin (IL)-12 expression, which is corrected by cyclooxygenase inhibitors. As exogenous arachidonic acid (AA) voids the NK blocking effect of PIBF, it is likely that PIBF acts before the level of the cyclooxygenase enzyme. Therefore in this study we investigated the effect of PIBF neutralizing antibody and simultaneous phospholipase A2 inhibitor quinacrine (Q) treatment on IL-12 production. METHODS: Pregnancy lymphocytes were treated with anti-PIBF antibody or lipopolysaccharide (LPS) as a positive control, in the presence or absence of Q. IL-12 expression by PBMC was detected by immunocytochemistry. RESULTS: Neutralization of PIBF as well as LPS treatment resulted in an increased IL-12 expression, which was corrected by simultaneous Q treatment. Pre-treatment of lymphocytes with progesterone prevented the stimulating effect of LPS on IL-12 production. CONCLUSION: Progesterone binding of the lymphocytes is followed by the release of PIBF that inhibits AA release. The subsequent block of prostaglandin synthesis reduces IL-12 production and results in a lowered cytotoxic NK activity, which may contribute to a normal pregnancy outcome. PMID- 12733590 TI - The herbal medicine Unkei-to stimulates the secretion of a cytokine-induced neutrophil chemoattractant, CINC/gro, in the rat ovarian cell culture. AB - PROBLEM AND METHOD OF STUDY: We investigated the ovulation-inducing effects of Unkei-to, a Japanese herbal medicine, in relation to the production of sex steroid hormones (17beta-estradiol and progesterone), cytokine-induced neutrophil chemoattractant (CINC/gro), interleukin-1beta (IL-1beta) and tumor necrosis factor alpha (TNF-alpha) in the rat ovarian cell culture. RESULTS: Unkei-to at a concentration of 100 microg/mL significantly stimulated the secretions of 17beta estradiol and progesterone (P < 0.01) in cultured whole ovarian dispersates. Unkei-to also enhanced the secretion of CINC/gro in a dose-dependent manner, and the secretions of CINC/gro increased significantly at concentrations of 10 and 100 microg/mL (P < 0.01). These stimulatory effects of Unkei-to on steroidgenesis and CINC/gro production are very similar to those of another Japanese herbal medicine, Toki-Shakuyaku-san. In addition, Unkei-to significantly (P < 0.01) enhanced the secretions of both IL-1beta and TNF-alpha, which are known to stimulate the secretion of CINC/gro in the ovulatory process, at concentrations of 10 and 100 microg/mL. The stimulatory effect of Unkei-to at a concentration of 100 microg/mL on IL-1beta/was significantly (P < 0.01) lower than that of Toki Shakuyaku-san, while the stimulatory effects of these two herbal medicines at a concentration of 100 microg/mL on TNF-alpha were similar. CONCLUSIONS: These results show that Unkei-to can stimulate ovarian steroidgenesis and the ovulatory process by inducing the secretion of CINC/gro with IL-1beta and TNF-alpha in vitro. Unkei-to has stimulatory effects on both steroidgenesis and the ovulatory process in the ovary as well as a stimulatory effect on the hypothalamus pituitary axis, and it may be useful for treating patients with ovulatory disorders. PMID- 12733591 TI - Effect of intravenous immunoglobulin treatment on the Th1/Th2 balance in women with recurrent spontaneous abortions. AB - PROBLEM: The way by which intravenous immunoglobulin (IvIg) acts to prevent immunlogically mediated recurrent spontaneous abortions (RSA) has not been clarified. In the present study, a possible effect of IvIg on the T helper cell (Th1/Th2) balance was investigated in abortions of either alloimmune or autoimmune abnormalities. METHOD OF STUDY: The study included 21 women treated with IvIg before conception because of a history of RSA characterized by alloimmune abnormalities (n = 15) or associated with anti-phospholipid antibodies (APA) (n = 6). Peripheral blood samples, collected before and 5 days after the first IvIg infusion, were stimulated, and Th1 and Th2 cells were detected by flow cytometric analysis using a combination of monoclonal antibodies against T-cell surface markers and intracellular interferon (IFN)-gamma and interleukin (IL)-4. The percentage of IFN-gamma-producing (Th1) and IL-4-producing (Th2) cells and the Th1/Th2 ratio were compared between pre- and post-infusion samples. RESULTS: A decrease of Th1 percentage in 66.6% of the cases and a concurrent Th2 percentage increase (47.61%) resulted in a decrease in the Th1/Th2 ratio in most of the cases (76.1%) (p < 0.01). Similar results were found in Group A (Th1/Th2 decreased in 60% of the cases, p < 0.05), while in Group B the effect of IvIg was not clear (Th1/Th2 increased in three and decreased in another three cases). CONCLUSION: Our finding suggests that IvIg administration in women with alloimmune RSA enhances Th2 polarization. This is not always the case with APA associated abortions. PMID- 12733592 TI - Potentiation of the maternal immune system may modify the apoptotic process in embryos exposed to developmental toxicants. AB - PROBLEM: We have previously shown that teratogen-induced embryonic maldevelopment may result from excessive apoptosis in affected organs, but the mechanisms underlying this process are not well understood. Here we investigate the ability of maternal immunopotentiation to affect the apoptotic process and its regulatory genes p53 and bcl-2 in embryos exposed to a teratogenic insult. METHOD OF STUDY: Potentiation of the immune system in pregnant females was performed with xenogeneic rat splenocytes or with granulocyte macrophage-colony stimulating factor (GM-CSF). The animals were exposed to cyclophosphamide (CP) and the reproductive performance in the various experimental groups was recorded. The level of apoptosis was assessed in the embryonic head and liver by TdT-mediated dUTP-biotin nick end labeling and fluorescence-activated cell sorter (FACS) analysis, while p53 and bcl-2 expression was evaluated by FACS and immunohistochemistry. RESULTS: In CP-treated females, a decrease in embryonic weight and an increase in the resorption rate and the percentage of embryos exhibiting head malformations were noted. These effects of CP were accompanied by the appearance of apoptotic cells in the head but not in the liver and an increased expression of p53 in embryonic organs, while bcl-2 expression was found to be decreased in the head and increased in the liver. Immunopotentiation with rat splenocytes or GM-CSF was shown to partially normalize the teratogenic effect of CP. It was also found to partially decrease the CP-induced apoptotic process and exhibited a tendency to normalize the expression of p53 and bcl-2 in the embryonic head and liver. CONCLUSION: Our results suggest a possible role for maternal immunopotentiation in protecting the embryo from teratogenic insults, possibly through regulation of the CP-induced apoptotic process and the expression of p53 and bcl-2. PMID- 12733593 TI - Effect of active immunization with the C-terminal 67-94 (R-28) region of human seminal plasma inhibin on the fecundity of adult male rabbits. AB - PROBLEM: To characterize the specificity of antibodies to the synthetic C terminal 67-94 (R-28) peptide of human seminal plasma inhibin (hSPI), and to examine the effect of active immunization on the fecundity of adult male animals. METHOD OF STUDY: The specificity of R-28-DT antibodies was tested using enzyme linked immunosorbent assay, immunohistochemical and immunofluorescence staining, and Western blotting. For fertility studies, adult male rats and rabbits were immunized and mated with females of the same strain. RESULTS: Rabbit antibodies to R-28-DT recognized native hSPI, as demonstrated by sperm agglutination in vivo and in vitro, on the sperm head by immunofluorescence staining, and in the columnar epithelial cells of the prostate by immunohistochemical staining. Immunization with the R-28-DT conjugate elicited a poor antibody response in male rats and their fecundity remained unaffected, while in male rabbits it elicited a good immune response with reduction in their fertility. CONCLUSION: R-28-DT antibodies recognized the native hSPI in the prostatic epithelium and agglutinated washed rat, rabbit, monkey, and human spermatozoa in vitro. Immunization of rabbits caused agglutination of spermatozoa resulting in a decrease in their fecundity. The conjugated R-28 peptide of hSPI offers promise as a male contraceptive. PMID- 12733594 TI - Antiprothrombin antibodies are associated with pregnancy loss in patients with the antiphospholipid syndrome. AB - OBJECTIVE: To document the clinical association between the history of pregnancy loss in patients with the diagnosis of primary or secondary antiphospholipid syndrome (APS) and the presence of different antiprothrombin antibody subtypes [immunoglobulin G (IgG), IgM and IgA] in a cohort of patients with APS. METHODS: Records of 170 female patients with primary APS, or APS secondary to systemic lupus erythematosus (SLE) or secondary to other autoimmune diseases were studied. RESULTS: In female APS patients with IgG antiprothrombin antibodies (n = 105) significant associations to pregnancy loss (p < 0.0001), early pregnancy loss (p < 0.0001) and a negative association to thrombocytopenia (p < 0.01) could be identified. In the group of patients with IgG antiprothrombin antibodies and at least one pregnancy (n = 84) a significant association with pregnancy loss (p < 0.005) and especially with early pregnancy loss (p < 0.0001) was demonstrated. No association with other immunoglobulin subtypes of antiprothrombin antibodies could be documented. In the subgroup of patients with primary APS and at least one pregnancy in the history, pregnancy loss (p < 0.005) and early pregnancy loss (p < 0.0001) were found to be highly associated with the presence of IgG antiprothrombin antibodies. IgG antiprothrombin antibodies represent the highest independent risk factor for pregnancy loss with an odds ratio of 4.5. There was no statistically significant association with venous or arterial thrombosis in all IgG antiprothrombin antibody positive patients. CONCLUSION: The results of this study document the association of IgG antiprothrombin antibodies with pregnancy loss and in particular early pregnancy loss in a large and well characterized cohort of patients. We would recommend routine testing for antiprothrombin antibodies in young female patients with APS. PMID- 12733595 TI - Glomerular immunoglobulin deposits induce glomerular inflammation in pregnant but not in non-pregnant rats. AB - PROBLEM: Does an inflammatory stimulus evoke a more intense inflammatory response in pregnant rats as compared with non-pregnant rats? METHOD OF STUDY: Non pregnant rats were injected with antibodies against the glomerular basement membrane (GBM), 14 days before pregnancy, to induce a subclinical glomerulonephritis. Part of the rats were rendered pregnant, the others remained non-pregnant throughout the experiment. Two experiments were performed: in experiment 1, pregnant and non-pregnant rats were killed at various intervals after the injection with antibody and parameters characteristic of a glomerular inflammation were evaluated using immunohistology on cryostat kidney sections and liver sections. In experiment 2, 24-hr urinary protein excretion was measured at various days after the injection in pregnant and non-pregnant rats. RESULTS: Experiment 1 revealed that a significant glomerular inflammation, as characterized by increased numbers of monocytes and LFA-1 positive cells per glomerulus, was only observed in pregnant rats with glomerulonephritis. Experiment 2 revealed that only pregnant rats with glomerulonephritis showed increased urinary protein excretion. CONCLUSION: The fact that glomerular inflammation coincides with proteinuria only in pregnant rats with glomerulonephritis, may suggest that these phenomena are causally related and promoted by the pregnant condition. PMID- 12733596 TI - Rhesus rhadinovirus infection in healthy and SIV-infected macaques at Tulane National Primate Research Center. AB - Rhesus rhadinovirus (RRV) infection was quantified in peripheral blood mononuclear cells (PBMC) from healthy and simian immunodeficiency virus (SIV) infected rhesus macaques (Macaca mulatta) at the Tulane National Primate Research Center and in a large collection of simian acquired immunodeficiency syndrome- (SAIDS)-associated lymphomas. Quantification of RRV load was performed by real time PCR using amplification primers specific for the RRV interleukin-6 homologue (RRV vIL-6). RRV infection was detected infrequently and at low levels in PBMC of randomly selected healthy animals. Examination of longitudinally collected PBMC from 22 SIV-infected animals throughout progression to SAIDS revealed similarly low RRV loads that sometimes increased with advancing disease. RRV infection was detected more frequently in the peripheral blood of SIV-infected animals than in healthy animals. Examination of SAIDS-associated lymphomas showed that RRV is rare within the tumor mass, likely representing infection in an occasional tumor infiltrating cell or contaminating blood. The results indicate that RRV infection in PBMC is not predictive of, and is apparently not required for, development of lymphoma or hyperplastic lymphadenopathy in SIV-infected animals at TNPRC. PMID- 12733597 TI - Naturally occurring fatal herpes simplex virus 1 infection in a family of white faced saki monkeys (Pithecia pithecia pithecia). AB - A family of three white-faced saki monkeys (Pithecia pithecia pithecia) died 48 96 hours after the onset of anorexia, nasal discharge, pyrexia and oral ulceration. One animal also had clonic seizures. Lesions found post-mortem consisted of oral and esophageal ulcers, hepatic and intestinal necrosis, meningoencephalitis and sporadic neuronal necrosis. Intranuclear inclusion bodies and syncytial cells were present in oral lesions and affected areas of liver. Herpes simplex virus 1 (HSV-1) was identified as the etiology of disease by virus isolation, polymerase chain reaction, or in situ hybridization in all three animals. Immunohistochemistry for detection of apoptotic DNA and activated caspase-3 showed significant levels of apoptosis in oral and liver lesions and occasional apoptotic neurons in the brain. These findings demonstrate the vulnerability of white-faced saki monkeys to HSV-1 and provide initial insight into the pathogenesis of fatal HSV-1-induced disease, indicating that apoptosis plays a significant role in cell death. PMID- 12733598 TI - Urinary steroids, FSH and CG measurements for monitoring the ovarian cycle and pregnancy in the chimpanzee. AB - Non-invasive methods for monitoring reproductive status of chimpanzee based on the measurement of urinary steroids and gonadotropins were examined. A typical pre-ovulatory urinary estrone conjugate (E1C) surge and post-ovulatory increase in pregnandiol glucuronide (PdG) were seen during the menstrual cycle. Urinary follicle stimulating hormone (FSH) showed two peaks over the infertile menstrual cycle. The earliest changes indicating pregnancy were a coincident rise in E1C and chorionic gonadotropin (CG) levels and a concomitant fall in FSH levels. Urinary PdG levels showed a prolonged rise. Urinary E1C in the pregnant chimpanzee was higher than during the menstrual cycle and increased with advancing gestation, with maximum levels occurring near term. In the case of stillbirth, E1C and CG levels from mid- through late-pregnancy were low and the prepartum progressive increase in E1C was not shown. The data presented here are of great practical value in captive breeding management of chimpanzees. PMID- 12733599 TI - Flow cytometric analysis of macaque whole blood for antigen-specific intracellular cytokine production by T lymphocytes. AB - We report here the standardized conditions for stimulation of macaque whole blood samples with various protein or peptide antigens, and production of significant intracellular levels of interferon gamma (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) in CD4+ as well as CD8+ T lymphocytes. We observed significantly higher levels of TNF-alpha compared with IFN-gamma in both CD4+ and CD8+ T lymphocytes from all the macaque whole blood samples stimulated with staphylococcal enterotoxin B (SEB) as an antigen. Similarly, when whole blood samples from rhesus macaques immunized with an HIV envelope peptide cocktail vaccine were stimulated with either the peptide cocktail or recombinant gp160, we observed production of significant levels of TNF-alpha by both CD4+ and CD8+ T lymphocytes. These results strongly support the utility of the whole blood cytokine flow cytometry methodology for determining antigen-specific immune responses of macaques in vaccine studies. PMID- 12733600 TI - Proliferative response of peripheral blood lymphocytes to mitogens in the owl monkey Aotus nancymae. AB - The new world primate Aotus sp. has been recommended by the World Health Organization as a model for evaluation of malaria vaccine candidates, given its susceptibility to experimental infection with the human malaria parasites Plasmodium falciparum and P. vivax. The present study examined the in vitro proliferative response of peripheral blood mononuclear cells (PBMCs) isolated from Aotus monkeys, utilizing a wide range of mitogens. Results presented herein demonstrate that the in vitro proliferative response of PBMCs from the Aotus sp. is quite variable from monkey to monkey for each of the mitogens assessed. PBMCs from the Aotus monkey exhibited a delayed kinetic proliferative response and, particularly, a different sensitivity to proliferation in response to various concentrations of Phytohemagglutinin-P and favin lectins, the phorbol ester Phorbol myristate acetate and the calcium ionophore ionomycin. Altogether, our findings are consistent with the conclusion that the in vitro proliferative response of PBMCs from the Aotus differ in their activation requirements compared with PBMCs from humans. PMID- 12733601 TI - Record review of baboons with histologically confirmed endometriosis in a large established colony. AB - Spontaneous endometriosis was diagnosed in 43 baboons over a 14-year period. Thirty-seven have died; five remain alive; one was sold and lost to follow-up. The average age at diagnosis was 17.2 years; 29 (67%) were between 12 and 21 years of age. Fifteen (35%) were diagnosed by biopsy and received surgical excision of the endometriotic tissue; four of these were identified during caesarian section, confirming one prior report of endometriosis in pregnant animals. Twenty-eight (65%) were diagnosed at or shortly preceding necropsy. When diagnosed by a palpable abdominal mass, there was a significantly greater likelihood the animal died or was killed as a result of complications of endometriosis. When diagnosis was at necropsy, there was a significantly greater likelihood that the animal died from causes unrelated to endometriosis. Early identification with surgical removal appears to provide a benefit for both survival and delivering offspring after diagnosis. In twenty-one baboons (49%), endometriosis affected multiple sites within the peritoneal cavity. In the remaining baboons, lesions were more localized. Ovarian involvement was seen in sixteen (37%) of these baboons. This paper is the first to describe significant ovarian involvement in baboons, previously considered a limitation of the usefulness of this species as an animal model. We also describe the first reported endometriosis seeding of an abdominal surgery scar in a baboon. Many of these baboons were middle aged, had few or no offspring, or had evidence of a long duration of uninterrupted menstrual cycles, consistent with risk factors for women. Endometriosis was an incidental finding in 17 (40%) of these baboons, consistent with previous reports of minimal endometriosis as a common asymptomatic finding in baboons and in women. Overall, endometriosis in baboons presents a spontaneously occurring animal model that shares important features with the disease in women and the rhesus macaque. PMID- 12733602 TI - Spontaneous ovarian tumors in twelve baboons: a review of ovarian neoplasms in non-human primates. AB - Twelve spontaneous ovarian tumors were found in the Southwest Foundation for Biomedical Research baboon colony. These included four granulosa cell tumors, three teratomas, two endometrioid carcinomas, one seromucinous cystadenofibroma, a cystic papillary adenocarcinoma, and an ovarian carcinoma. Age was a pre disposing factor. With one exception, the tumors of surface epithelial- and sex cordstromal origin occurred in baboons over 17 years of age. The exceptional animal was 7 years of age when a malignant granulosa cell tumor with Sertoli cell differentiation was identified. The two endometrioid tumors, which were found in 17- and 30-year-old animals, were both associated with endometriosis. In contrast, the teratomas, which are tumors of germ cell origin, were found in younger animals, i.e. 17 years of age or younger. One case of an ovarian carcinoma with metastases was observed in a 6-month-old infant. Cases of spontaneous ovarian tumors from the literature are reviewed. PMID- 12733604 TI - Clinical utility of resistance testing. PMID- 12733603 TI - Ovarian stimulation of marmoset monkeys (Callithrix jacchus) using recombinant human follicle stimulating hormone. AB - To reduce the number of animals required for controlled studies of marmoset oocytes and early embryos, a superovulation protocol was developed for the common marmoset. Females were given up to 50 i.u./day recombinant human follicle stimulating hormone (FSH)--(r-hFSH) for 6 days. Ovaries were visualized by a modified laparoscopic technique and follicular aspiration was performed using a needle and suction apparatus inserted directly through an otoscope speculum. The number of follicles + ovulation points (+/- S.E.) was 2.9 (+/- 0.2) in controls and 14.1 (+/- 1.6; P < or = 0.001) in the 50 i.u. r-hFSH per day animals. Oocytes, typically at the germinal vesicle stage at collection, extruded a first polar body within 26 hours. In vitro fertilization was performed and embryos developed to the hatched blastocyst stage (34%). With many high quality oocytes and the ability to synchronize cycles, the marmoset is a valuable primate model for examining nuclear reprograming and early embryonic events. PMID- 12733605 TI - Resistance tests: what do clinical trials tell us? AB - Few routine laboratory tests have been evaluated by randomised controlled trials. Nevertheless, a number of such studies have been undertaken for genotypic and phenotypic HIV drug resistance assays. They are all based on a structure whereby patients are randomised to receive resistance tests or none (standards of care) at the time of acute viral failure, with the primary end points being the proportion with undetectable viral load at a time point 1224 weeks following randomisation. In other words, these studies assess the benefit afforded to clinicians by resistance data. We show that these studies vary in their results, and identify the potential factors associated with such variation. Although most studies demonstrate a clinical benefit of resistance testing, we argue that the opportunity for undertaking further such studies is probably gone and that effort should be focused on identifying more precisely the factors which determine the most cost-effective use of these expensive tests. PMID- 12733606 TI - Resistance to protease inhibitors. AB - Resistance to HIV-1 protease inhibitors (PIs) limits the benefits of highly active antiretroviral therapy (HAART) and complicates the selection of appropriate salvage therapy for a growing number of patients. Initial patterns of drug resistance depend on the PI used in treatment, viral subtype and level of drug exposure. Impaired replication capacity of PI-resistant viruses contributes to the maintenance of immunological stability despite ongoing virus replication. However, the accumulation of additional mutations leads eventually to higher levels of drug resistance and immunological decline. Resistance may be overcome by pharmacological enhancement of plasma PI levels, but newer drugs that are active against PI-resistant viruses are needed. PMID- 12733607 TI - The impact of HIV-1 subtype on the clinical response on HAART. AB - Highly active antiretroviral therapy (HAART) has led to a reduction in HIV related mortality and morbidity. Most patients who have benefited from HAART are infected with HIV-1 subtype B, which predominates in Western Europe, the USA and the rest of the industrialised world. However, most HIV-infected people live in sub-Saharan Africa, Asia and Eastern Europe. In these areas, subtypes other than B are responsible for the epidemic of HIV-1 infection. This review focuses on the clinical significance of HIV-1 infection with a non-B subtype. The increase in availability of HAART to developing countries together with the large number of HIV-1-infected immigrants being treated in the industrialised world means that data on the clinical response to therapy for non-B HIV-1 infections are becoming of greater practical relevance. If antiretroviral agents, which generally target subtype B, are less efficacious in non-B infections, this will have major clinical implications for therapeutic strategies. Data on drug susceptibility, clinical response and the development of resistance in non-B HIV-1 subtypes are discussed here. PMID- 12733608 TI - A virological perspective on resistance in highly treatment-experienced patients. AB - An increased number of HIV-infected patients have experienced most or all available antiretroviral drugs. or these patients, building a salvage or rescue regimen is difficult. Resistance to highly active antiretroviral therapy (HAART) makes the clinical decision process complex. However, growing understanding of HIV dynamics and resistance patterns to HAART may help physicians make more rational decisions when planning treatments in highly experienced patients. Viral resistance mutations may have favourable effects in salvage treatment by reducing viral fitness and pathogenicity. Similarly, interactions between mutations may possibly increase susceptibility to other antiretroviral drugs. PMID- 12733609 TI - Building the best regimen for highly treatment-experienced patients. AB - The use of highly active antiretroviral therapy (HAART) has dramatically reduced the mortality and morbidity associated with HIV infection. However, drug resistance remains the most important reason for virological failure and rapidly limits treatment options. Virus resistant to all three major antiretroviral drug classes is now reported even in primary seroconverters. In this article, the options for the management of highly treatment-experienced patients with multiple drug resistance are considered. Options discussed include whether to continue failing therapy, stop treatment in the hope of the virus reverting to wild type, the role of resistance testing in this setting, the impact of reduced viral fitness and the use of mega- and giga-HAART. PMID- 12733610 TI - Comparing the financial risk of bed-day and DRG based pricing types using parametric and simulation methods. AB - The extent of random financial risk involved in the Finnish bed-day and Diagnosis Related Groups (DRG) based hospital pricing systems were estimated and compared using parametric and simulation methods. DRG based payment schemes were found to provide significantly better protection against financial risk for municipalities, but municipality's size was the main determinant of financial risk. Small municipalities should use longer contracts between hospitals or form bigger purchaser-organisations for risk pooling. In addition, the current risk management system proved to be ineffective in decreasing the random variation in total costs. PMID- 12733611 TI - IS/IT the prescription to enable medical group practices attain their goals. AB - The US spends significantly more money as a percentage of GDP on health care than any other OECD country and more importantly, this amount is anticipated to increase exponentially. In this high cost environment, two important trends have occurred: (1) the movement to managed care, and (2) large investments in Information Systems/Information Technology (IS/IT). Managed care has emerged as an attempt to provide good quality yet cost effective health care treatment. Its implications are not well discussed in the literature while, its impact on different types of medical group practices is even less well understood. The repercussions of the large investments in IS/IT on the health care sector in general and on the medical group practice in particular, although clearly of importance, are also largely ignored by the literature. This study attempts to address this significant void in the literature. By analyzing three different types of group practices; an Independent Practice Association (IPA), a Faculty Practice and a Multi Specialty Group Practice in a managed care environment during their implementation of practice management/billing systems, we are able to draw some conclusions regarding the impacts of these two central trends on health care in general as well as on the medical group practice in particular. PMID- 12733612 TI - Appraising the economic efficiency of cancer treatment: an exploratory analysis of lung cancer. AB - This paper tests whether the measured cost-effectiveness of treating different subgroups of an incident population of lung cancer patients differs significantly and, by implication, whether the provision of care to these patients is tolerably efficient in economic terms. Data from administrative records and Registry follow up on 544 non-small cell lung cancer patients diagnosed at a single NCI designated Comprehensive Cancer Center are used to conduct the empirical analysis. The main results show statistically significant differences in cumulative costs and patient outcomes across subgroups differing by disease stage and treatment modality. These findings imply that the delivery of lung cancer care is inefficient. Substantive and methodological implications are discussed. PMID- 12733613 TI - Analytical methods for calculating the capacity required to operate an effective booked admissions policy for elective inpatient services. AB - In the UK, hospitals are being encouraged to introduce booked admissions policies for elective inpatient services whereby patients are given a date for hospital admission months in advance rather than being put on a waiting list and then informed of their admission date at short notice. We address the question of what level of capacity is required to operate such a system if cancellations of booked elective patients are to be kept to a low level. Methods are presented for quantifying the day to day variation in bed demand due to emergency admissions, patient initiated cancellations and variable lengths of stay amongst patients. PMID- 12733614 TI - Client involvement in simulation model building: hints and insights from a case study in a London hospital. AB - This paper describes the collaborative process of building a simulation model in order to understand patient waiting times in an accident and emergency department. The purpose is to explore the issues that arise when involving clients, in this case health care professionals, in the process of model building. The study background and a detailed account of the modelling process are presented. The focus of the paper is on some general themes that can be discovered running through the process. These offer some tentative insights into client involvement in system dynamics modelling, in particular its application in health care. Given this study's first promising results, further collaborative studies are encouraged. PMID- 12733615 TI - Integrated healthcare networks' performance: a growth curve modeling approach. AB - This study examines the effects of integration on the performance ratings of the top 100 integrated healthcare networks (IHNs) in the United States. A strategic contingency theory is used to identify the relationship of IHNs' performance to their structural and operational characteristics and integration strategies. To create a database for the panel study, the top 100 IHNs selected by the SMG Marketing Group in 1998 were followed up in 1999 and 2000. The data were merged with the Dorenfest data on information system integration. A growth curve model was developed and validated by the Mplus statistical program. Factors influencing the top 100 IHNs' performance in 1998 and their subsequent rankings in the consecutive years were analyzed. IHNs' initial performance scores were positively influenced by network size, number of affiliated physicians and profit margin, and were negatively associated with average length of stay and technical efficiency. The continuing high performance, judged by maintaining higher performance scores, tended to be enhanced by the use of more managerial or executive decision-support systems. Future studies should include time-varying operational indicators to serve as predictors of network performance. PMID- 12733616 TI - Impulsiveness, impulsive aggression, personality disorder, and spousal violence. AB - Impulsiveness has become a key concept in thinking about the determinants of violence and aggression. In this study of spouse abusers, the relationship between impulsiveness, impulsive aggression, and physical violence is confirmed. Impulsiveness and impulsive aggression have significant correlations with physical aggression. Impulsiveness and impulsive aggression are also correlated with measures of Borderline Personality Disorder and Antisocial Personality Disorder. In addition, the measures of Borderline and Antisocial Personality Disorder (PD) are significantly correlated with physical aggression. The violent and non-violent groups differed on impulsive aggression and on Borderline Personality Disorder. A partial replication of Tweed and Dutton's findings (1998) revealed sub-groups of high- and low-violence men. The high-violence group was very different from the low-violent and the non-violent groups. The high-violence group had higher pathology scores on all clinical scales, except Mania, of the Personality Assessment Inventory. These findings have implications for violence prediction and for treatment of violent men. PMID- 12733617 TI - Different paths to death row: a comparison of men who committed heinous and less heinous crimes. AB - Part of the answer to violent crime prevention is to understand the route that those who have committed violent crimes have traveled in order to find ways to guide others from the road leading to such violence. An investigation of the lifelong personal and environmental factors affecting 37 men who were executed in 1997 focuses on distinctions between men in two categories based on heinousness of violent crime. The study aimed to identify risk factors and events that preceded the violent event and to compare the constellation of variables of the men who committed particularly heinous murders characterized by extreme rage and brutality with those whose crimes and criminal histories were characterized mostly by property crimes without intentional harm to people. Descriptive results suggest differences between the two groups of men related to 19 variables and the emergence of two diverse profiles of risk factors and life experiences. PMID- 12733618 TI - The political economy of urban homicide: assessing the relative impact of gender inequality on sex-specific victimization. AB - This research examines the ways in which the changing political economy of urban areas has contributed differently to the homicide victimization rates of females and males across US cities. Recent research, while relatively limited, has presented disparate results regarding the effect of gender inequality on urban sex-specific victimization. Our work further explores this relationship by taking into account relative gender disparities in income, education, labor market opportunities, and politics in an examination of sex-specific homicide victimization in 1990. Key to this current investigation is the evaluation of feminist and lifestyle arguments that suggest that structural gender inequality has a unique effect on female victimization. Overall, our findings reveal gender inequality to be a significant predictor of both male and female urban homicide. While these findings suggest mixed support for theoretical arguments regarding gender inequality, further analyses reveal significant differences in specific types of gender inequality on victimization patterns across genders. These additional results highlight the need for greater attention toward both methodological and theoretical issues when examining the interconnections between gender, political economy, and violence in research. PMID- 12733619 TI - Structural antecedents of aggravated assault: exploratory assessment of female and male victimization. AB - This article examines whether the social structural factors predicting violence against women are different from those predicting violence against men. Using sex specific, aggravated assault rates from Florida counties (n = 60), this regression analysis tests three principal explanations of violent victimization: routine activities, social disorganization, and gender inequality. Although initially some difference in the predictive factors for male and female aggravated assault rates emerged, a test of the equality of regression coefficients revealed no "real" significant differences. Despite this finding, it remains important to assess the influence of societal factors on rates of violent victimization. The national trend indicates that male violent victimization is declining and female violent victimization is relatively stable. It is important to understand why this is the case. PMID- 12733620 TI - Vicarious traumatization, secondary traumatic stress, and burnout in sexual assault and domestic violence agency staff. AB - This study investigated three occupational hazards of therapy with trauma victims: vicarious trauma and secondary traumatic stress (or "compassion fatigue"), which describe therapists' adverse reactions to clients' traumatic material, and burnout, a stress response experienced in many emotionally demanding "people work" jobs. Among 101 trauma counselors, client exposure workload and being paid as a staff member (vs. volunteer) were related to burnout sub-scales, but not as expected to overall burnout or vicarious trauma, secondary traumatic stress, or general distress. More educated counselors and those seeing more clients reported less vicarious trauma. Younger counselors and those with more trauma counseling experience reported more emotional exhaustion. Findings have implications for training, treatment, and agency support systems. PMID- 12733621 TI - The relative effects of intimate partner physical and sexual violence on post traumatic stress disorder symptomatology. AB - This study examined the relative effects of intimate partner physical and sexual violence on Post-Traumatic Stress Disorder (PTSD) symptomatology. Severity of physical and sexual violence as well as PTSD severity were assessed in a sample of 62 help-seeking battered women. The results of this study were consistent with prior research, finding significant and positive relationships between physical and sexual violence as well as sexual violence and PTSD symptoms. In order to further clarify these relationships, the unique effects of sexual violence on PTSD were examined after controlling for physical violence severity. Results indicated that sexual violence severity explained a significant proportion of the variance in PTSD severity beyond that which was already accounted for by physical violence severity. These findings have important implications for mental health and social service professionals who work with battered women. PMID- 12733622 TI - The effects of relationship aversive female partner behavior on attributions and physiological reactivity of verbally aggressive and non-aggressive males. AB - The present study assessed the effects of aversive female partner behavior on cognitive attributions and physiological reactivity in verbally aggressive and non-aggressive college males (N = 39). Participants were presented four audiotaped vignettes which depicted hypothetical dating situations in which the female's behavior was relationship aversive or non-relationship aversive. Participants' physiological reactivity (i.e., systolic blood pressure, diastolic blood pressure, and heart rate) was obtained before and after hearing each vignette. Attributional responses were obtained following the presentation of all vignettes. Relationship aversive partner behavior was expected to produce greater increases in attributional and physiological reactivity than non-relationship aversive partner behavior. Additionally, verbally aggressive males were expected to demonstrate greater negative intent and responsibility attributions and evidence greater physiological reactivity for situations involving relationship aversive partner behavior than were non-aggressive males.As hypothesized, results showed that relationship aversive partner behavior produced greater increases in systolic and diastolic blood pressure than did non-relationship aversive partner behavior. Results also showed that verbally aggressive males evidenced significantly greater negative attributions to relationship aversive partner behavior than did non-aggressive males. The potential interaction between physiological reactivity and attributions in explaining males' verbally aggressive behavior toward their female partners is discussed. PMID- 12733623 TI - Fungal species are new records for Turkey. PMID- 12733624 TI - The cell-mediated immune reaction in the cutaneous lesion of chromoblastomycosis and their correlation with different clinical forms of the disease. AB - Chromoblastomycosis is a fungal infection caused by dematiaceous fungi inducing skin lesions of difficult treatment and of frequent recurrence. The objective of the present investigation was to characterize cell-mediated tissue reactions in the skin in cases of Chromoblastomycosis using histopathology and immunocytochemistry methods and to correlate them with different clinical forms of Chromoblastomycosis. Biopsies from 19 patients were stained with HE and Giemsa, and serial sections were immunohistochemically stained using CD45RO, CD20, CD4, CD8, CD68, CD1a, CD34, IL4, IL10, TNF-alpha and IFN-gamma antibodies. A quantitative and semiquantitative analysis of the cell subsets and cytokines in the inflammatory infiltrates was performed by counting ten high-power fields (400x). The cutaneous lesion presented as verrucous plaque (n = 15) or erythematous atrophic plaque (n = 4). We observed two types of tissue reaction: A) a granulomatous reaction with a suppurative granuloma with several fungi cells in the cutaneous lesion presenting as verrucous plaque; B) a granulomatous reaction with a tuberculoid granuloma with few fungi cells in the cutaneous lesion presenting as atrophic plaque. The data obtained suggest that patients with lesion presented as verrucous plaque have a type Th2 immunological response, while patients with lesion presented as erythematous atrophic plaque have a type Th1 response. PMID- 12733625 TI - Fluorescent method for studying the morphogenesis and viability of dermatophyte cells. AB - The dermatophyte Microsporum canis is commonly isolated from human and animal infection. The morphogenesis of this fungus was studied during its developmental stages through the fluorescent method Fluorescein Diacetate and Ethidium Bromide. To this end, 50 microl dermatophyte suspension were transferred onto cellophane wrapping esterilized discs (2.5 cm of diameter) placed over the surface of Sabouraud dextrose agar on Petri dishes and incubated at 25 degrees C for 30 days. Every 60 minutes during the first 24 hours and every 12 hours for next 29 days, one disc was transferred onto glass slide, covered with equal volumes of freshly prepared fluorescein diacetate (FDA) and ethidium bromide (EB) solution, mounted with a coverslip and incubated in the dark for 30 minutes, at 25 degrees C. Each preparation was then examined on a fluorescent microscope. M. canis presented well defined growth stages: (1) tumescence of cells; (2) germination; (3) development of hyphae; (4) production of conidia and (5) tumescence and formation of arthroconidiae. Using the fluorescent method, non viable cells showed a light bright red coloration and viable cells presented green fluorescence. The principal morphological changes have occurred between the 3rd until the 18th day of culture. The method is very useful to demonstrate the dermatophyte growth stages as well as the perfect differentiation between viable and non viable cells. PMID- 12733626 TI - Germination, viability and clearance of Stachybotrys chartarum in the lungs of infant rats. AB - Observing that the conidia of Stachybotrys chartarum can germinate in the lung of infant rats, it became important to ascertain whether an infection can ensue. Viable conidia of S. chartarum were instilled into the lungs of 4 and 14 day-old rat pups. Germination was observed frequently in the lungs of 4 day-old but rarely in the 14 day-old pups. In the 4 day-old pups, pulmonary inflammation with hemorrhagic exudates was observed and resulted in about 15% mortality rate compared to 0% for the controls instilled with phosphate buffered saline. Acute neutrophilic inflammation and intense interstitial pneumonia with poorly formed granulomas observed three days following exposure were associated with fungal hyphae and conidia. The surviving experimental pups showed significantly slower weight gain for seven days. Dilution plating and quantitative PCR analysis were used to follow total fungal load in the rat pups lung homogenates. In the 4 day old rat pups viable fungi decreased rapidly and were less than 1% by day seven. Similarly, fungal DNA decreased exponentially and was only 0.03% by fourteen days after exposure. However, 14 day-old rat pups showed neither the lethal effects of exposures to viable conidia of S. chartarum nor the slower weight gain, and the fungal load decreased even more rapidly. We conclude that S. chartarum conidia can initially germinate and form hyphae but even in the immature rat pups do not establish an effective infection, although a very limited persistence cannot be excluded. PMID- 12733627 TI - Aplanes braunii (Phylum Oomycota) causes mortality in Anabas cobojius. AB - The fresh water fish Anabas cobojius (Hamilton-Buchanan 1822), was found to be parasitised by Aplanes braunii de Bary 1888. It is the first report of Aplanes braunii to cause disease on any fish. A. braunii is a virulent parasite that caused cotton-wool disease involving the skin, fins and muscles of A. cobojius. The hyphae also penetrated deep into the tissues of the infected fish and all of them ultimately died. PMID- 12733628 TI - Mycoflora of tuber surface of white yam (Dioscorea rotundata Poir) and postharvest control of pathogens with Bacillus subtilis. AB - Bacillus subtilis (Enrenberg) Cohn was investigated for its antagonistic properties against surface mycoflora of yam (Dioscorea rotundata Poir) tubers in storage. Yam tubers inoculated with a spore suspension of B. subtilis in potato dextrose broth using a knapsack sprayer showed a drastic reduction in the range and number of mycoflora, including pathogens of the tuber surface in contrast to the control tubers, during the five-month storage period in a traditional yam barn. However, B. subtilis maintained a high frequency of occurrence during the same period. Botryodiploidia theobromae Pat, Fusarium moniliforme Wollen and Reink., Penicillium sclerotigenum Yamamoto, and Rhizoctonia sp. were displaced completely on the treated tubers. The antagonism of B. subtilis was so effective that the normal tuber surface mycoflora was greatly reduced throughout the storage period of five months by a simple initial application of the antagonist. PMID- 12733629 TI - Fungi associated with food and feed commodities from Ecuador. AB - Freshly harvested soybean, rice and corn from farms and corn-based pelleted feeds were collected from ranches from the coastal and mountain regions in Ecuador during 1998, and assessed for fungal contamination. The most prevalent fungi on pelleted feed were Aspergillus flavus and Fusarium graminearum. The prevalent fungi recovered from soybean were F. verticillioides, F. semitectum, Aspergillus flavus and A. ochraceus. In rice, F. oxysporum was the most prevalent toxigenic fungal species recorded, followed by F. verticillioides and A. flavus. In corn, F. verticillioides was the most prevalent fungus isolated in both the coastal and mountain regions, with high isolation frequencies of A. flavus and A. parasiticus at the coast. Based on the toxigenic species recovered, ochratoxin A may pose a contamination risk for soybean. A higher probability of aflatoxin contamination of corn was found in the coastal samples compared to those of the mountain region, while a risk of fumonisin contamination of corn exists in both regions. PMID- 12733630 TI - Water availability affects the growth, accumulation of compatible solutes and the viability of the biocontrol agent Epicoccum nigrum. AB - Growth of the biocontrol fungus Epicoccum nigrum was more sensitive to ionic solute water stress (NaCl) than non-ionic (glycerol) on potato dextrose-based media at -0.5, -3.0 and -5.5 MPa water potentials. Subsequent physiological manipulation of growth of E. nigrum in glycerol-modified media to -3.0 MPa water potential resulted in a significant increase in the accumulation of compatible solutes in both mycelial liquid cultures and spores, but no enhanced accumulation of the desiccation protectant trehalose, when compared to unmodified media (-0.5 MPa). The main solute accumulated was glycerol, followed by arabitol. In temporal studies over 20 days maximum accumulation of glycerol occurred in 5-d old cultures with water stressed cultures having 250x greater amounts than those from unmodified medium. The arabitol content was also higher in mycelium and spores produced under water stress. The difference was maximum after 15 days growth. Glucose content decreased over time in mycelial colonies but increased in spores. The germination of conidia from the two treatments was similar, regardless of compatible solute content, even at -9.25 MPa water potential stress. However, germ tube extension was significantly increased at this water potential level. The production of E. nigrum spores at -3.0 MPa water potential resulted in improved survival when stored fresh at 4 and 25 degrees C. However, freeze-drying severely affected the viability of spores produced on both media (-0.5 or 3.0 MPa). Accumulation of compatible solutes may assist the fungus in better ecological competence and establishment in the phyllosphere, where water availability is often limited. PMID- 12733631 TI - Isozyme polymorphism and virulence of Indian isolates of the rice sheath blight fungus. AB - Inadequate information about the genetic structure of the polyphagous Rhizoctonia solani has made sheath blight resistance breeding a difficult task. To assess the variability in the Indian populations of sheath blight fungus, 18 isolates were collected from different rice growing regions of India and analyzed for virulence and electrophoretic profiles of 13 isozymes. The virulence spectrum of all 18 isolates was examined on susceptible IR50 and tolerant Swarnadhan varieties, based on which the isolates could be grouped as highly virulent, moderately virulent or avirulent. A total of 11 enzyme systems with 153 electrophoretic phenotypes were applied to characterize the genetic variation among the isolates. Cluster analyses based on isozyme patterns resulted in one major cluster comprising 16 virulent isolates, with two avirulent isolates loosely linked to this at 0.13 similarity. Isozyme systems of esterases (both alpha and beta) and 6 phosphogluconic dehydrogenase could be used to fingerprint the individual isolates. PMID- 12733632 TI - Immunocytochemical localization of stachylysin in Stachybotrys chartarum spores and spore-impacted mouse and rat lung tissue. AB - Stachylysin is a proteinaceous hemolytic agent that is produced by Stachybotrys chartarum. Stachylysin was found, using immunohistochemical and immunocytochemical methods, to be localized in S. chartarum spores/mycelia primarily in the inner wall suggesting that it is constitutively produced. Spores instilled in mouse or rat lung tissues resulted in granuloma formation, which showed the highest stachylysin concentration in the inner wall of the spore and near the spore, with less at distance indicating that it had diffused out from the spore. The in vitro high stachylysin producing strain (58-06) was also highest in vivo, based on immunohistochemistical staining. More stachylysin was observed in the mouse lung tissue at 72 h than at 24 h indicating that production/release is a relatively slow process. The localization of stachylysin in macrophage phagolysosomes suggests that these cells may be involved with hemolysin inactivation. This would be consistent with what is known about asp hemolysin produced by Aspergillus fumigatus. PMID- 12733633 TI - Histological, immunohistochemical and morphometric changes in lung tissue in juvenile mice experimentally exposed to Stachybotrys chartarum spores. AB - Stachybotrys chartarum is an important toxigenic fungus often associated with chronically wet cellulose-based building materials. The purpose of this study was to evaluate some histological, immunohistochemical and morphometric changes in mouse lung tissues exposed intratracheally to either 50 microl of 1.4 x 10(6) S. chartarum spores (< or = 35 ng toxin/kg BW), isosatratoxin-F (35 ng/kg BW), 50 microl of 1.4 x 10(6) Cladosporium cladosporioides spores, or 50 microl saline. Exposure of lung tissues to S. chartarum or C. cladosporioides spores resulted in granuloma formation at the sites of spore impaction. Some of the lung tissues impacted by S. chartarum spores also showed erythrocyte accumulation in the alveolar air space, dilated capillaries engorged with erythrocytes, and hemosiderin accumulation at spore impaction sites, which were features not noted in the C. cladosporioides-spore treated animals. Immunohistochemistry revealed reduced collagen IV distribution in lung granulomas in S. chartarum-treated animals especially at 48 and 72 hr post-exposure compared to that in lungs of mice with C. cladosporioides-spore induced granulomas. Quantitative analysis of pooled S. chartarum and C. cladosporioides spore impacted lungs revealed significant depression (P < 0.05) of alveolar air space from 71.4 +/- 6.1% in untreated animals to 56.04 +/- 6.1% in the S. chartarum- and 60.24 +/- 5.5% in the C. cladosporioides-spore treated animals. It also revealed that alveolus air space in S. chartarum treated animals declined significantly from 63.74 +/- 3.1% at 12 hr post-exposure to 42.94 +/- 7.9% at 72 hr post-exposure and was increased to 54.84 +/- 5.2% at 96 hr post-exposure. Alveolus air space in C. cladosporioides treated animals also decreased significantly from 64.84 +/- 7.1% at 12 hr exposure to 54.94 +/- 5.4% at 48 hr post-exposure and was increased to 64.64 +/- 10.1% at 96 hr post-exposure. It also revealed significant (P < 0.05) alveolar accumulation of erythrocytes from 1.24 +/- 1.4% in the untreated animals to 3.44 +/- 1.5% in the pooled S. chartarum spore treated animals. Erythrocyte abundance in S. chartarum treated animals increased significantly (P < 0.001) from 2.14 +/- 1.7% at 12 hr post-exposure to 5.54 +/- 1.5% at 72 hr and 4.94 +/- 1.4% at 96 hr post-exposure. These results further reveal that exposure to S. chartarum spores elicit tissue responses in vivo significantly different from those associated with exposure to pure trichothecene toxin and to spores of a non toxigenic fungus. PMID- 12733634 TI - Isolation and toxigenicity of Aspergillus fumigatus from moldy silage. AB - Thirty-nine silage samples were collected from various silos on Terceira Island in the Azores. Samples were examined for the presence of total fungi, and isolates of Aspergillus fumigatus were analyzed for their ability to produce fumitremorgens B and C, fumigaclavines B and C, and gliotoxin. Thirty-four silage samples (87%) were contaminated with fungi, and A. fumigatus was isolated from 27 samples (69%). Samples that were taken from the surface of silos had significantly higher populations of both total fungi and A. fumigatus than did samples taken from the middle of silos. Analysis of 27 A. fumigatus isolates (one representing each positive sample) showed that 59.3% produced fumitremorgen B; 33.3% produced fumitremorgen C; 29.6% produced fumigaclavine B; 7.4% produced fumigaclavine C; and 11.1% produced gliotoxin. Fifty-two percent of the isolates produced multiple toxins, and 25.9% did not produce any of these toxins. Gliotoxin and fumigaclavine C were always produced in combination with other toxins. Because of the demonstrated potential of these A. fumigatus isolates to produce mycotoxins, it is important to properly construct and manage silos to prevent their contamination with A. fumigatus. PMID- 12733635 TI - Screening toxicity study in young carp (Cyprinus carpio L.) on feed amended with fumonisin B1. AB - Fumonisin B1 (FB1) is one of several mycotoxins produced by Fusarium moniliforme, a major fungal pathogen of corn and widely spread throughout the world. FB1 produces a wide range of biological effects, some of which are specific for particular organs or species and some are common to all investigated animals. In this study we have evaluated subchronic toxicosis features in young carp (Cyprinus carpio L.) exposed to 0.5 and 5.0 mg FB1 kg(-1) body weight for 42 days through nutritionally balanced diet. During the trial we observed loss of body weight in both treated groups, together with higher incidence of infective bacterial dermatological lesions erythrodermatitis cyprini (Aeromonas salmonicida subsp. nova) in the group treated with the higher FB1 dose. Several hematological parameters (erythrocyte count, platelet count) and serum chemical concentrations (creatinin, total bilirubin) and activities (aspartate aminotransferase, AST and alanine aminotransferase, ALT) were greater in the fumonisin treated groups than in the control group. Our results indicate that long-term dietary exposure to 0.5 and 5.0 mg FB1 kg(-1) body weight is not lethal to young carp, but can produce adverse physiological effects. These findings also suggest that primary target organs of FB1 in the carp are kidney and liver, as it has already been observed in other animal species tested. Specifically changed red blood cell- parameters reveal that FB1 probably causes erythrocyte membrane defect or interferes with carp's respiratory process. PMID- 12733636 TI - Chromosome painting in the African four-striped mouse Rhabdomys pumilio: detection of possible murid specific contiguous segment combinations. AB - Fluorescence in-situ hybridization was used to construct a comparative chromosome map between the laboratory mouse, Mus musculus and the African four-striped mouse, Rhabdomys pumilio. A high degree of homology between the species was detected using both FISH and G-banding. Ten mouse chromosomes (2-4, 7, 14-16, 18, 19 and the X) were retained as chromosomal arms or intact chromosome blocks. Six mouse chromosome painting probes that correspond to mouse autosomes 5, 6, 8, 11, 12 and 13, produced double signals; the remaining four painting probes (1, 9, 10 and 17) hybridized to three or more R. pumilio chromosomes respectively. In total, the 20 mouse chromosome paints revealed 40 segments of conserved synteny in the R. pumilio genome. Most of the mouse chromosomes that produced single signals in R. pumilio have previously been shown to be conserved in the Black and Norwegian rats and the Chinese hamster. Eight contiguous segment associations appear to be R. pumilio specific, two were shared by R. pumilio and the Black and Norwegian rats, but to the exclusion of the Chinese hamster. Our data suggest that mouse chromosomes 1, 10, and 17 have undergone extensive rearrangements during genome evolution in the murids and may be useful markers for enhancing our understanding of the mode and tempo of chromosome evolution in rodents. PMID- 12733637 TI - Lampbrush chromosomes of the chaffinch (Fringilla coelebs L.). AB - The seven macrochromosomes of the chaffinch (Fringilla coelebs L.) are described in their lampbrush form. The relative lengths of bivalents, the positions and arrangements of chromosomal regions with lateral loops of similar length and appearance, as well as the positions of protein bodies and loops of peculiar morphology have been defined and mapped, so that each of the seven lampbrush macrobivalents may be identified in oocytes from every individual of the species. This morphological analysis has been supplemented by determining the positions of certain loci and objects that are specifically and consistently labelled after immunostaining or fluorescence in-situ hybridization with defined molecular probes. PMID- 12733638 TI - B-A interchanges are an unlikely pathway for B chromosome integration into the standard genome. AB - One of the conceivable evolutionary pathways of a parasitic B chromosome is its integration into the host genome through translocation to an A chromosome. To investigate this possibility, we analyze here the nature, meiotic behavior and genetic effects of a spontaneous interchange between a medium-sized autosome and a B chromosome, found in one male caught in a Moroccan population of the grasshopper Eyprepocnemis plorans and the offspring of controlled crosses with six different females. Most metaphase I cells analyzed (115 out of 118) showed a trivalent with chiasmata in both the interstitial and pairing segments, which predicted about half of genetically unbalanced spermatozoa. The analysis of 234 embryos sired by this male on six females showed the lethality of some meiotic products paralleled to a decrease in egg fertility (0.541 +/- 0.051, compared to the 0.879 +/- 0.017 shown by females mated to standard males). These results suggest that the cost of the B-A interchange on host fitness, in terms of gametic inviability, highly diminishes the possibility of frequency increase for the interchange to reach a polymorphic status, which is the first and indispensable step to reach fixation and thus integration of the B chromosome DNA into the host genome. PMID- 12733639 TI - Caenorhabditis elegans Ce-rdh-1/rad-51 functions after double-strand break formation of meiotic recombination. AB - During meiotic prophase 1, homologous recombination is accompanied by dynamic chromosomal changes. The Ce-rdh-1/rad-51 gene is the only bacterial recA-like gene in the nematode C. elegans genome. Upon depletion of Ce-rdh-1/rad-51 using the RNA interference method, abnormal 'kinked' chromosomes can be observed in mature oocytes at diakinesis, whereas synapsis between homologous chromosomes during the pachytene stage is normal. Following fertilization, Ce-rdh-1/rad-51 depleted embryos die early in embryogenesis, and their nuclei exhibit abnormal chromosome fragments and bridges. From epistasis analyses with Ce-spo-11 defective mutant and ionizing radiation, it is indicated that Ce-rdh-1/rad-51 functions after double-strand break (DSB) formation of meiotic recombination. Under the Ce-chk-2 defective condition, whose meiotic synapsis and meiotic recombination between homologous chromosomes are completely inhibited, the Ce-rdh 1/rad51 is normally expressed in the gonadal cells. Moreover, it seems that exogenous DSBs in the Ce-chk-2 defective nuclei at the pachytene stage can be repaired between sister chromatids in a Ce-rdh-1/rad-51-dependent manner. These results indicate that Ce-rdh-1/rad51 functions after both endogenous and exogenous DSB formation during meiosis, but not as 'pairing centers' for meiotic synapsis. PMID- 12733641 TI - A probe generated by chromosome microdissection, useful for analyzing Y chromosome evolution in Old World monkeys. AB - We isolated a DNA probe, designated MMDYZ1, using a chromosome microdissection technique from the Y chromosome of the Rhesus monkey. The probe obtained from eight whole Y chromosomes shows higher specificity for the Y short arm of the Rhesus monkey, which consists totally of constitutive heterochromatin. Two microclones (MMY#3 and MMY#4) were constructed from the Y-specific primary PCR products. Sequence analysis of these two microclones revealed that both were essentially identical to each other and the sizes were 870 and 686 bp, respectively. From alignment analysis using the Genbank database of primates, the alphoid DNA has the highest affinity with the probe. However, the total composition of this probe has extremely high homology with the Y short arm of the Rhesus monkey, as demonstrated by fluorescence in-situ hybridization (FISH). Comparative FISH-mapping disclosed that this DNA-sequence cluster was located at extremely different sites on the Y chromosome in several species of the Old World monkey. Accordingly, this probe seems to be a high-quality tool, now established for the first time, for investigating Y chromosome evolution of the Old World monkey. PMID- 12733640 TI - RNA interference is required for normal centromere function in fission yeast. AB - In plants, animals and fungi, active centromeres are associated with arrays of repetitive DNA sequences. The outer repeats at fission yeast (Schizosaccharomyces pombe) centromeres are heterochromatic and are required for the assembly of an active centromere. Components of the RNA interference (RNAi) machinery process transcripts derived from these repeats and mediate the formation of silent chromatin. A subfragment of the repeat (dg) is known to induce silencing of marker genes at euchromatic sites and is required for centromere formation. We show that the RNAi components, Argonaute (Ago1), Dicer (Dcr1) and RNA-dependent RNA polymerase (Rdp1), are required to maintain silencing, lysine 9 methylation of histone H3 and association of Swi6 via this dg ectopic silencer. Deletion of Ago1, Dcr1 or Rdp1 disrupts chromosome segregation leading to a high incidence of lagging chromosomes on late anaphase spindles and sensitivity to a microtubule poison. Analysis of dg transcription indicates that csp mutants, previously shown to abrogate centromere silencing and chromosome segregation, are also defective in the regulation of non-coding centromeric RNAs. In addition, histone H3 lysine 9 methylation at, and recruitment of Swi6 and cohesin to, centromeric repeats is disrupted in these mutants. Thus the formation of silent chromatin on dg repeats and the development of a fully functional centromere is dependent on RNAi. PMID- 12733642 TI - Spatiotemporal sites of DNA replication in macro- and micronuclei of the ciliate Paramecium caudatum. AB - Spatiotemporal sites of DNA replication in macro- and micronuclei of the ciliated protozoan Paramecium caudatum were analyzed by confocal laser scanning microscopy following incorporation of the thymidine analogue BrdU and indirect immunofluorescence. In the macronucleus, replication sites were localized to numerous small domains and scattered throughout the nucleoplasm. This pattern persisted during all periods of the S phase. A single constant pattern with discrete replication foci was also observed in the micronucleus. No obvious differences were seen between the two kinds of nuclei. Pulse-chase-pulse double labeling experiments with two thymidine analogues (CldU and IdU) revealed that dispersed sites of replication were activated at different times during the S phase and a replication site takes about 2h to complete replication in the macronucleus. When cells were labeled by BrUTP to examine transcriptional activity in the two kinds of nuclei, incorporation of BrUTP into the macronucleus occurred throughout the cell cycle, whereas there was no detectable RNA synthesis in the micronucleus. From these findings, we conclude that, despite large differences in structure and function of macro- and micronuclear genomes, both nuclei show a similar replication pattern with discrete subnuclear foci scattered throughout the nucleoplasm at all times during the S phase. PMID- 12733643 TI - Karyological and genetic variation in Middle Eastern lacertid lizards, Lacerta laevis and the Lacerta kulzeri complex: a case of chromosomal allopatric speciation. AB - Karyological (standard and C, Ag-NOR and Alu-I banding methods) and mtDNA analyses (cytochrome b and 12S rRNA) were conducted on specimens from eight allopatric populations of the Lacerta kulzeri complex. Parallel analyses were performed for comparison on Lacerta laevis specimens. Karyological and molecular studies support the morphological and ethological evidence indicating the specific separation between Lacerta laevis and Lacerta kulzeri. In the Lacerta kulzeri complex, chromosomal analysis substantiated an interpopulation differentiation roughly along a north-south trend, mainly regarding the sex chromosome morphology and heterochromatin. The cytochrome b and 12S rRNA gene analyses showed minor genetic differences that were considerably smaller than those commonly found in genetically isolated populations. The L. kulzeri populations from Barouk, Druze and Hermon show a mean genetic distance that, in other saurians, characterises subspecies. The conditions found in L. laevis and L. kulzeri are reminiscent of King's model of chromosomal primary allopatry and support the hypothesis that in these lacertid lizards chromosome variations can become fixed before the accumulation of the genetic mutations. PMID- 12733645 TI - Base oxidation at 5' site of GG sequence in double-stranded DNA induced by UVA in the presence of xanthone analogues: relationship between the DNA-damaging abilities of photosensitizers and their HOMO energies. AB - UVA contributes to skin cancer by solar UV light. Photosensitizers are believed to play an important role in UVA carcinogenesis. We investigated the mechanism of DNA damage induced by photoexcited xanthone (XAN) analogues (XAN, thioxanthone [TXAN] and acridone [ACR]), exogenous photosensitizers, and the relationship between the DNA-damaging abilities and their highest occupied molecular orbital (HOMO) energies. DNA damage by these photosensitizers was examined using 32P labeled DNA fragments obtained from the p53 tumor suppressor gene. Photoexcited XAN caused DNA cleavage specifically at 5'-G of the GG sequence in the double stranded DNA only when the DNA fragments were treated with piperidine, suggesting that DNA cleavage is due to base modification with little or no strand breakage. With denatured single-stranded DNA, the extent of XAN-sensitized photodamage was decreased. An oxidative product of G, 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo dGuo), was formed by photoexcited XAN, and the 8-oxo-dGuo formation was decreased in single-stranded DNA. TXAN and ACR induced DNA photodamage as did XAN, although the order of DNA-damaging ability was XAN > TXAN > ACR. These findings suggest that photoexcited XAN analogues induce nucleobase oxidation at 5'-G of GG sequence in double-stranded DNA through electron transfer. The HOMO energies of these photosensitizers, estimated from ab initio molecular orbital (MO) calculation, decreased in the following order: XAN > TXAN > ACR. Extents of DNA damage increased exponentially with the HOMO energies of XAN analogues. This study suggests that DNA-damaging abilities of photosensitizers can be estimated from their HOMO energies. PMID- 12733644 TI - Topically applied vitamin C and cysteine derivatives protect against UVA-induced photodegradation of suprofen in ex vivo pigskin. AB - Exposure of the nonsteroidal anti-inflammatory drug suprofen (SUP) to UV radiation results in the formation of radicals, reactive oxygen species (ROS), photodecarboxylated products and photoadducts with biomacromolecules. Using an ex vivo pigskin explant model, we investigated whether topical coapplication of the water-soluble antioxidants vitamin C (Lascorbic acid, ASC), N-acetyl-L-cysteine (NAC) or L-cysteine ethylester (CYSET) with SUP reduced ultraviolet A (UVA) induced decomposition of SUP. UVA-induced changes in antioxidant bioavailability in the stratum corneum and epidermis were also studied. Epidermal bioavailability of SUP in sham-irradiated pigskin increased 2.2- to 4.1-fold after the lowest antioxidant doses (P < 0.05). As compared with no applied antioxidant, increasing doses of all tested antioxidants resulted in increased levels of SUP and decreased levels of photoproducts (P < 0.05). A maximal protection against SUP photodegradation of 70% was found after an ASC dose of 1 micromol/cm2; these values were 60% for a NAC dose of 10 micromol/cm2 and 50% for a CYSET dose of 5 micromol/cm2. Skin antioxidant levels increased with increasing applied dose (P < 0.05); the bioavailability of CYSET was approximately three-fold lower than that of ASC and NAC. UVA exposure resulted in 30-50% consumption of the topically applied ASC or NAC in the stratum corneum, whereas CYSET was not consumed. In conclusion, the topically applied water-soluble antioxidants ASC, NAC and CYSET protect against UVA-induced decomposition of SUP by scavenging radicals and ROS. Coapplication of these antioxidants may therefore be an effective way to reduce or prevent the phototoxic effects of SUP in vivo. PMID- 12733646 TI - Photomutagenic properties of terfenadine as revealed by a stepwise photostability, phototoxicity and photomutagenicity testing approach. AB - Administration of the second-generation antihistamine, terfenadine, is sometimes associated with photosensitivity and other skin reactions. To obtain information on its photoreactivity, we used a stepwise experimental approach involving tests for photostability, phototoxicity (PT) (mouse fibroblast cell line [3T3] neutral red uptake [NRU] test) and photomutagenicity (with standard Ames salmonella tester strains TA98, TA100 and TA102). Terfenadine was not phototoxic to cultured mammalian cells under the conditions used (i.e. 5000/161 mJ cm(-2) UVA-UVB). Natural sunlight and UV radiations caused considerable drug decomposition and formation of several photoproducts. Addition of the irradiated terfenadine solution (i.e. a mixture of photoproducts) to the tester did not significantly increase background mutation frequency. Irradiation of terfenadine coplated with the TA102 strain induced a clear-cut photomutagenic response, the magnitude of which was dependent upon the precursor compound concentration and the UV dose (212/7 to 339/11 mJ cm(-2) UVA-UVB). These findings demonstrate that in vitro terfenadine is photomutagenic in absence of PT. Further in vitro and in vivo studies are therefore needed to provide an adequate safety assessment of the photochemical genotoxicity--carcinogenicity potential of terfenadine. In the meantime, patients should be advised to avoid excessive exposure to sunlight. PMID- 12733647 TI - Single-molecule fluorescence lifetime and anisotropy measurements of the red fluorescent protein, DsRed, in solution. AB - Fluorescence lifetime and anisotropy measurements were made on the red fluorescent protein (DsRed) from tropical coral of the Discosoma genus, both at single-molecule and bulk concentrations. As expected from previous work, the fluorescence lifetime of DsRed in solution is dependent on laser power, decreasing from an average fluorescence lifetime in the beam of about 3.3 ns at low power (3.5 ns if one extrapolates to zero power) to about 2.1 ns at 28 kW/cm2. At the single-molecule level, exciting with 532 nm, 10 ps laser pulses at 80 MHz repetition rate, DsRed particles entering the laser beam initially have a lifetime of about 3.6 ns and convert to a form having a lifetime of about 3.0 ns with a quantum yield of photoconversion on the order of 10(-3) (calculated in terms of photons per DsRed tetramer). The particles then undergo additional photoconversion with a quantum yield of roughly 10(-5), generating a form with an average lifetime of 1.6 ns. These results may be explained by rapid photoconversion of one DsRed monomer in a tetramer, which acts as an energy transfer sink, resulting in a lower quantum yield for photoconversion of subsequent monomers. Multiparameter correlation and selective averaging can be used to identify DsRed in a mixture of fluorophores, in part exploiting the fact that fluorescent lifetime of DsRed changes as a function of excitation intensity. PMID- 12733648 TI - Photooxidation of 2'-deoxyguanosine 5'-monophosphate in aqueous solutions. AB - Mass spectrometric and ultraviolet absorption spectral evidence are presented for the assignment of structures to three products detected in the reaction mixtures formed upon the photolysis of aqueous solutions of the nucleotide 2' deoxyguanosine 5'-monophosphate (dGMP) with light of wavelengths >270 nm. The products for which structures are assigned are spiroiminodihydantoin 2' deoxyribonucleotide (1), 2,2-diamino-4-([2-deoxy-5-monophosphate-beta-D-erythro pentofuranosyl]amino)-5-(2H)-oxazolone (oxazolone 2'-deoxyribonucleotide, 2) and 2-amino-5-([2-deoxy-5-monophosphate-beta-D-erythro-pentofuranosyl]amino)-4H imidazol-4-one (imidazolone 2'-deoxyribonucleotide, 3). These results, when combined with mechanistic data presented in an earlier communication, provide support for the proposal that the irradiation of dGMP with UVB light leads to the formation of singlet oxygen. The UV absorption spectral properties of the imidazolone make this product a reasonable candidate to rationalize the autosensitization of dGMP degradation reported in the earlier communication. PMID- 12733649 TI - Trehalose-induced changes of the ethidium hydration shell detected by time resolved fluorescence. AB - The fluorescence lifetime of ethidium bromide (EB), a widely used fluorescent dye, has been monitored in water solution versus trehalose concentration in order to learn how the presence of the disaccharide modifies the hydration of EB, which is used in this study as a model probe. The interest in trehalose, a naturally occurring bioprotector (osmolyte), stems from its ability to stabilize biomolecules under stress situations. The observed effects of trehalose on EB fluorescence properties have been compared with those induced by sucrose and glucose. Lifetime measurements have been performed by frequency domain fluorometry (2-40 MHz), and from the analysis of their changes versus sugar concentration, the EB-sugar interaction parameters have been obtained. The effect of trehalose on the EB decay in glasses has also been studied both by exploring the heterogeneity of lifetime decay and by single-molecule imaging. Trehalose appears to be more efficient in changing the EB fluorescence parameters, such as the emission lifetime, and it leads to a degree of heterogeneity higher than that induced by the other sugars. When EB is embedded in trehalose glasses, the heterogeneity of the emission and of the bleaching time is further enhanced. PMID- 12733650 TI - Immunotoxic effects of cis-urocanic acid exposure in C57BL/6N and C3H/HeN mice. AB - Exposure to ultraviolet radiation results in increased levels of intradermal cis urocanic acid (cUCA) and alters cutaneous immunity by interfering with processing and presentation of antigen by Langerhans cells. Reports on effects of systemic immunotoxicity with 30 day cUCA exposure in laboratory rodents include thymic atrophy, thymic hypocellularity and decreased T-cell-mediated immunity; however, immune effects of single exposure or 5 day cUCA administration, which may better mimic human exposures, are poorly defined. The present study initially evaluated immune effects of single, 5 day, and 4 week cUCA exposure in C57BL/6N mice. Single administration of intradermal cUCA resulted in decreased splenocyte phagocytosis that persisted for 30 days after cUCA exposure. Five day consecutive cUCA exposure decreased numbers of phenotypically mature CD4(+)CD8(-) and CD4( )CD8(+) (single positive) thymocytes, increased CD4(+)CD8(+) (double positive) immature thymocytes and increased splenocyte proliferation. Prolonged cUCA exposure (4 weeks) caused profound thymic hypocellularity and splenic hypercellularity and increased splenic macrophage chemiluminescence. Because of this apparent sensitivity of C57BL/6N mice to cUCA, thymic hypocellularity was compared between C57BL/6N and C3H/HeN mice dosed with cUCA, and was found to be more pronounced in the C57BL/6N strain. These results are an extension of previous conclusions on immune modulation caused by cUCA in the spleen and thymus. Further, the observed variation in sensitivity between the mouse strains is consistent with known genetic susceptibility of these strains to the immunomodulatory effects of exposure to sunlight. PMID- 12733651 TI - Prompt and delayed nonsteroidal anti-inflammatory drug-photoinduced DNA damage in peripheral blood mononuclear cells measured with the comet assays. AB - Nonsteroidal anti-inflammatory drug (NSAID)-photoinduced DNA damage in human peripheral blood mononuclear cells measured using the alkaline comet assay is presented. Whereas Tiaprofenic Acid-photoinduced DNA damage was promptly induced (i.e. observed at relatively low radiation doses), Ketoprofen-photoinduced DNA damage was delayed. This prompt and delayed effect is observed with UVA (320-400 nm), UVB (290-320 nm) and solar-simulated radiation and is attributed to the different photochemical properties of NSAID. The results from these experiments, carried out in living cells, confirm the speculations of NSAID-photoinduced DNA damage brought up by the many experiments conducted in solution. PMID- 12733652 TI - Sensitivity of cyanobacterial antenna, reaction center and CO2 assimilation transcripts and proteins to moderate UVB: light acclimation potentiates resistance to UVB. AB - We tested the hypothesis that cyanobacterial cells have sufficient acclimation potential to tolerate UVB when it is applied in a natural quantum ratio to growth photosynthetically active radiation (PAR). We grew Synechococcus under 50 (Low) or 300 (High) micromol PAR m(-2) x s(-1) and then exposed the cells to 0.125 (Low) or 0.75 (High) micromol UVB m(-2) x s(-1). The PAR:UVB quantum ratios were near natural for both the Low-PAR:Low-UVB and the High-PAR:High-UVB treatments, but UVB was in excess of typical aquatic PAR:UVB for Low-PAR:High-UVB treatments. The cellular light history determined the UVB responses of Synechococcus. High PAR cells initially had fewer cpc transcripts encoding phycocyanin, lower phycocyanin content, and more psbAII/AIII transcripts encoding the D1:2 photosystem II (PSII) protein isoform. Higher PAR potentiated them to tolerate an appropriate UVB level without short-term inhibition of PSII or growth. Low-PAR cells rapidly altered psbAII/AIII and cpc gene expression and tolerated appropriate Low UVB. Low-PAR:High-UVB cells, in contrast, suffered short-term inhibition of PSII and growth. In all treatments UVB induced transient loss of cpc transcripts, possibly to free resources for psbAII/AIII expression, which is important for UVB resistance. The drop in cpc transcripts was not part of a general shock response because rbcL transcript pools were stable upon UVB exposure. PMID- 12733653 TI - The combination of benzo[a]pyrene and ultraviolet A causes an in vivo time related accumulation of DNA damage in mouse skin. AB - Polycyclic aromatic hydrocarbons, including benzo[a]pyrene (BaP), are ubiquitous environmental carcinogens. BaP is metabolized in vivo to reactive intermediates that become covalently bound to DNA and form BaP-DNA adducts, an initial event in carcinogenesis. Ultraviolet A (UVA) synergizes with BaP to significantly enhance genetic damage and accelerate carcinogenic processes. This study was initiated to investigate in vivo cellular changes related to carcinogenesis induced by repeated exposures to BaP plus UVA. Simulated chronic exposure to an environmental carcinogen and sunlight was conducted through biweekly topical application of BaP followed 2 h later by UVA exposure over a 10 week period. BaP diol epoxide (BPDE)-DNA adducts were measured in vivo by immunohistochemistry using an anti-BPDE-DNA monoclonal antibody. Oxidative DNA damage was measured by the detection of 8-hydroxy-2'-deoxyguanosine (8-OHdG) formation using high performance liquid chromatography. Alterations in the cell cycle that were relevant to carcinogenesis were revealed by changes in p53, as identified in vivo using a polyclonal anti-p53 antibody. We found that cells containing BPDE-DNA adducts and nuclear p53 expression significantly increased between 2 and 10 weeks of BaP-UVA treatment, whereas neither BPDE-DNA adducts nor significant changes in p53 were observed in untreated skin. Using regression analysis, oxidative 8-OHdG damage also showed a parallel increase over 2-10 weeks (r = 0.80). These results indicate that genetic damage caused by exposures to BaP plus UVA accumulates with time and increases the potential for inductive events leading to carcinogenesis and tumor formation. PMID- 12733654 TI - Sensitivity and depth penetration of continuous wave versus frequency-domain photon migration near-infrared fluorescence contrast-enhanced imaging. AB - The development of near-infrared fluorescent contrast agents and imaging techniques depends on the deep penetration of excitation light through several centimeters of tissue and the sensitive collection of the re-emitted fluorescence. In this contribution, the sensitivity and depth penetration of various fluorescence-enhanced imaging studies is surveyed and compared with current studies using continuous wave (CW) and frequency-domain photon migration (FDPM) measurements with planar wave illumination of modulated excitation light at 100 MHz and area collection of reemitted fluorescent light using a previously developed modulated intensified charge-coupled device camera system. Fluorescence was generated from nanomolar to micromolar solutions of indocyanine green (ICG) in a 100 microL volume submerged at 1-4 cm depths in a 1% Liposyn solution to mimic tissue scattering properties. Enhanced depth penetration and sensitivity are achieved with optimal filter rejection of excitation light, and FDPM rejection of background light is not achieved using CW methods. We show the ability to detect as few as 100 fmol of ICG from area illumination of 785 nm light (5.5 mW/cm2) and FDPM area collection of 830 nm fluorescent light generated from 3 cm below the phantom surface. The lowered noise floor of FDPM measurements enables greater sensitivity and penetration depth than comparable CW measurements. PMID- 12733655 TI - Targeting cells that overexpress the epidermal growth factor receptor with polyethylene glycolated BPD verteporfin photosensitizer immunoconjugates. AB - Photoimmunotherapy was introduced two decades ago but has been studied infrequently in vivo and is virtually untested clinically. Progress has been limited because high-quality, well-characterized photosensitizer immunoconjugates (PICs) have been difficult to make. Here, we describe the development of an innovative conjugation method for producing water-soluble PICs that are free of insoluble aggregates and free of unacceptable amounts of noncovalently associated photosensitizer impurities. The method exploits two procedures previously untried in this research area. First, a small number of antibody lysines (<3 per antibody) are polyethylene glycolated (PEGylated) using a 10 kDa branched polyethylene glycol (PEG), which dramatically enhances PIC solubility and reduces PIC aggregation. Second, a 50% dimethyl sulfoxide-50% aqueous two-solvent system is used to prevent photosensitizer aggregation and noncovalent interactions. These measures allow efficient covalent linkage of the photosensitizer BPD Verteporfin (BPD) to antibody lysines, thorough purification of the resulting PICs (verified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis), maintenance of PIC antigen-binding activity (verified by cellular binding-uptake assays) and reduction of nonspecific cellular uptake (e.g. macrophage capture) of the PICs. Loading levels could be varied controllably in the range < or = 11 BPD/antibody. PICs of the C225 anti-epidermal growth factor receptor (EGFR) chimeric monoclonal antibody killed EGFR-overexpressing A-431 cells photodynamically but did not significantly affect EGFR-negative NR6 cells. Although fluorescence measurements demonstrated that the PICs were quenched by as much as an order of magnitude compared with free BPD, an impressive 90% reduction in A-431 cell viability was achieved using 20 J/cm2 of 690 nm light after a 40 h incubation with the C225 PICs. The results suggest that PEGylated BPD-C225 PICs merit further investigation in animal models of EGFR-overexpressing cancers. PMID- 12733656 TI - The effect of UVB irradiation on ferritin subunit synthesis, ferritin assembly and Fe metabolism in cultured canine lens epithelial cells. AB - Ferritin is a multimeric protein consisting of heavy and light chains assembled in different tissue-specific ratios, which can protect cells from oxidative stress by storing reactive iron (Fe). Because the lens is constantly exposed to UV irradiation, we studied its effects on ferritin synthesis and Fe metabolism in cultured lens epithelial cells with and without ascorbic acid (Asc). UVB caused a large increase in accumulation of newly synthesized ferritin chains; this increase was additive to that induced by Asc. In contrast to the Asc-induced increase in Fe storage, Fe storage in ferritin was unaltered by UVB. Although UVB increased accumulation of newly synthesized ferritin chains, total ferritin levels were unaltered. In contrast, Asc, which induced a quantitatively similar increase in accumulation of newly synthesized ferritin chains, doubled the total amount of ferritin. Because UVB did not change Fe storage in ferritin or the size of the labile Fe pool, it was hypothesized and then determined that these newly synthesized chains did not assemble into functional holoferritin. Numerous studies detail the effects of various treatments on de novo ferritin synthesis; however, this study provides a cautionary note regarding the conclusions of such studies in the absence of data indicating assembly of functional ferritin molecules. PMID- 12733657 TI - Energy transfer evidence for in vitro and in vivo complexes of Vibrio harveyi flavin reductase P and luciferase. AB - Conservation of energetically "expensive" metabolites is facilitated by enzymatic intra- and intermolecular channeling mechanisms. Our previous in vitro kinetic studies indicate that Vibrio harveyi reduced nicotinamide adenine dinucleotide phosphate-flavin mononucleotide (NADPH-FMN) oxidoreductase flavin reductase P (FRP) can transfer reduced riboflavin 5'-phosphate (FMNH2) to bacterial luciferase by direct channeling. However, no evidence has ever been reported for such an FMNH2 channeling between these two enzymes in vivo. The formation of a donor-acceptor enzyme complex, stable or transient, is mandatory for direct metabolite channeling between two enzymes regardless of details of the transfer mechanisms. In this study, we have obtained direct evidence of in vitro and in vivo FRP-luciferase complexes that are functionally active. The approach used is a variation of a technique previously described as Bioluminescence Resonance Energy Transfer. Yellow fluorescence protein (YFP) was fused to FRP to generate an active FRP-YFP fusion enzyme, which emits fluorescence peaking at 530 nm. In comparison with the normal 490 nm bioluminescence, an additional 530 nm component was observed in both the in vitro bioluminescence from the coupled reaction of luciferase and FRP-YFP and the in vivo bioluminescence from frp gene-negative V. harveyi cells that expressed FRP-YFP. This 530 nm bioluminescence component was not detected in a control in which a much higher level of YFP was present but not fused to FRP. Such findings indicate an energy transfer from the exited emitter of luciferase to the FRP component of the luciferase-FRP-YFP complex. Hence, the formation of an active complex of luciferase and FRP-YFP was detected both in vitro and in vivo. PMID- 12733658 TI - UV doses of young adults. AB - Since 1986, people have been informed that they get about 80% of their lifetime ultraviolet (UV) dose by the age of 18. This belief originated from the mathematical conclusion that diligent use of sunscreens (sun protection factor 15 or higher) during the first 18 years of life would reduce the lifetime incidence of nonmelanoma skin cancers by 78%. These data were misconstrued to mean that individuals also got about 80% of their lifetime dose of UV by the age of 18 (linear relationship). However, these calculations were based on the incidence of nonmelanoma skin cancers being related to the square of the UV dose. Careful analysis of UV exposure data shows that Americans actually get less than 25% of their lifetime UV dose by the age of 18. This finding also appears to be true worldwide because Australia, UK and The Netherlands report a similar UV exposure pattern. UV-initiated damage early in life can be promoted by subsequent exposures to progress into tumors later in life. For example, the nonmelanoma skin cancer, squamous cell carcinoma, is dependent on the cumulative UV dose. Thus, a better educational approach for reducing skin cancers would be to instruct fair-skinned individuals to protect themselves throughout their lives from being exposed to too much UV radiation. PMID- 12733659 TI - Quantitative genetic analysis of larval life history traits in two alpine populations of Rana temporaria. AB - We estimated genetic and maternal variance components of larval life history characters in alpine populations of Rana temporaria (the common frog) using a full-sib/half-sib breeding design. We studied trait plasticity by raising tadpoles at 14 or 20 degrees C in the laboratory. Larval period and metamorphic mass were greater at 14 degrees C. Larval period did not differ between populations, but high elevation metamorphs were larger than low elevation metamorphs. Significant additive variation for larval period was detected in the low altitude population. No significant additive variation was detected for mass at metamorphosis (MM), which instead displayed significant maternal effects. Plasticity in metamorphic mass of froglets was greater in the high altitude population. The plastic response of larval period to temperature did not differ between the populations. Evolution of metamorphic mass is likely constrained by lack of additive genetic variation. In contrast, significant heritability for larval period suggests this trait may evolve in response to environmental change. These results differ from other studies on R. temporaria, suggesting that populations of this broadly distributed species present substantial geographic variation in the genetic architecture and plasticity of tadpole life history traits. PMID- 12733660 TI - Conservation of chromosomal location of nucleolus organizer in American marsupials (Didelphidae). AB - The distribution and expression of nucleolus organizer regions (NORs) were analyzed in seven species of marsupials representative of the three karyotypes (2n = 14, 18 and 22) found in the American family Didelphidae. Analyses comprised silver-staining of NORs and fluorescence in situ hybridization with an rDNA probe. In addition to confirming the variability in number and distribution of NORs in Didelphidae, we demonstrated the conserved location of NORs on one autosome pair in the three karyotypes. In Monodelphis domestica (2n = 18), the NOR on the X chromosome was not inactivated in females. PMID- 12733661 TI - Isolation and analysis of a new hopper hAT transposon from the Bactrocera dorsalis white eye strain. AB - A new hopper element belonging to the hAT transposon family was isolated from the white eye mutant strain of the Oriental fruit fly, Bactrocera dorsalis. Using the original hopper element sequence from the wild type Kahuku strain as a template, the new hopper was isolated by inverse and direct PCR. Nucleotide sequence analysis reveals a 3131 bp element with terminal and subterminal inverted repeat sequences, an 8 bp duplicated insertion site, and a conceptual translation yielding a single uninterrupted 650 amino acid open reading frame. The white eye hopper has structure more consistent with function than the Kahuku element, indicating that hopper is not an ancient relic. The hopper element remains distantly related to other known hAT elements including those from insects, and presently it is most similar to Activator-related elements discovered in the human genome. DNA hybridization studies indicate, however, that elements closely related to hopper exist in another bactrocerid species, the melonfly, B. cucurbitae. PMID- 12733663 TI - On the relative roles of faster-X evolution and dominance in the establishment of intrinsic postzygotic isolating barriers. AB - The modern theory of speciation assigns a prominent role to the recessivity of genetic incompatibilities in the two rules of speciation, namely Haldane's rule and the 'large X effect', and considers that the contribution of faster evolution of the X versus the autosomes to those patterns is generally of relatively minor importance. By extending Turelli and Orr's previous analysis of the model of two locus Dobzhansky-Muller incompatibilities, I first show that when the X and the autosomes evolve at the same rate, the two dominance parameters involved in that model are not equally important for the declaration of a large X effect, but that the degree of recessivity of homozygous-homozygous incompatibilities is the major determinant for such a declaration. When the X evolves faster than the autosomes, the model obviously predicts that the importance of both dominance parameters will progressively vanish. It is then of importance to obtain estimates of the relative evolutionary rate of X-linked incompatibility loci. Several different procedures to obtain such estimates from the perspective of the large X effect are suggested. The application of the appropriate test to the only suitable data from Drosophila hybridizations so far available leads to the conclusion that the X actually evolves at least 2.5 times faster than the autosomes, as far as hybrid male sterility determinants are concerned, thus making dominance considerations absolutely irrelevant. Notwithstanding the necessity of further tests, the relative roles currently assigned to faster-X evolution and dominance in the theory of speciation should be revised, giving due prominence to faster-X evolution, at least for hybrid male sterility in the genus Drosophila. PMID- 12733662 TI - Cytogenetics of the genus Arvicanthis (Rodentia, Muridae). 3. Comparative cytogenetics of A. neumanni and A. nairobae from Tanzania. AB - The African rats of the genus Arvicanthis have been widely studied during recent years to clarify species boundaries and phylogenetic relationships. The wide chromosomal variability of the genus has been highlighted in several studies, with each accepted species characterised by its individual karyotypes and others being revealed as cryptic species. In the present paper we report the karyotype and the C- and G-banding patterns of the two species A. nairobae and A. neumanni from seven localities of Tanzania, an area of the range poorly studied. The two karyotypes were compared to that of A. niloticus, which is considered to be primitive. The karyotype of A. neumanni is characterised by 2n = 53-54 and NFa = 62. This karyotypic variability depends on a widespread Robertsonian polymorphism. The karyotype of A. nairobae shows 2n = 62 and NFa = 78; it diverges from that of A. niloticus through one reciprocal translocation, five inversions and three heterochromatic additions. The comparison with the karyotypes of other species of the genus showed that A. neumanni belongs to the east African lineage (with A. abyssinicus, A. blicki, A. niloticus), while A. nairobae is closer to the central and the west African representatives which were all previously under the name of A. niloticus (ANI-2, ANI-3, ANI-4). The distribution of A. nairobae in east Africa opens new scenarios in the biogeographical pattern of evolution of the genus. PMID- 12733664 TI - Variability on the dot chromosome in the Drosophila simulans clade. AB - A recent study suggested that recent nuclear gene introgression between Drosophila simulans and D. mauritiana may have obscured efforts to estimate the phylogeny of the species of the D. simulans clade, which includes these two species and D. sechellia. Here, we report sequence variation of an intron of the eyeless gene in this species group. This gene should introgress freely between these species because it is not linked to any known barriers to gene exchange. We have also reevaluated levels of sequence divergence among species in this clade, noting differences between loci in regions of low recombination (as in all chromosome 4 loci) relative to other loci. Overall, none of the data analyzed were consistent with recent introgression exclusively between D. simulans and D. mauritiana. PMID- 12733665 TI - Moderate-density molecular maps of Eucalyptus urophylla S. T. Blake and E. tereticornis Smith genomes based on RAPD markers. AB - Moderate-density molecular maps were constructed for the genomes of Eucalyptus urophylla S. T. Blake and E. tereticornis Smith using RAPD markers and an interspecific cross between the two species. One hundred and eighty-three primers were employed to generate 245 and 264 parent-specific markers in E. urophylla and E. tereticornis, respectively, as well as 49 parent-shared markers. The normally segregating markers, including 208 (84.9%) specific to maternal E. urophylla, 175 (66.3%) to paternal E. tereticornis, and 48 shared by both parents, were used for framework map construction for each parental species. For maternal E. urophylla, the linkage map consisted of 23 linkage groups, 160 framework markers, and 60 accessory markers, defining a total map distance of 1504.6cM and an average interval of 11.0 +/- 8.07 cM. For paternal E. tereticornis, the linkage map contained 23 linkage groups, 126 framework markers, and 92 accessory markers, defining a total map distance of 1035.7 cM and an average interval of 10.1 +/- 7.23 cM. Genome length was estimated at 1585.7 and 1507.5 cM for E. urophylla and E. tereticornis, respectively, indicating map coverage of 94.9 and 68.7% of the corresponding genomes. Construction of such maps will be valuable for quantitative trait loci (QTLs) detection, marker-assisted selection (MAS), comparative mapping, and whole genome based fingerprint characterization in Eucalyptus breeding programs. PMID- 12733666 TI - Larval polytene chromosomes of black flies (Simulium) from Thailand. I. Comparison among five species in the subgenus Gomphostilbia enderlein. AB - Larval polytene chromosome maps of Simulium (G.) asakoae and S. (G.) sp. g in the ceylonicum-group and S. (G.) angulistylum, S. (G.) decuplum and S. (G.) siamense in the batoense-group of the subgenus Gomphostilbia from Thailand are presented. These species have three pairs of chromosomes (2n = 6). Light stained centromeric bands were observed in the chromosomes of S. (G.) asakoae, S. (G.) sp. g, S. (G.) decuplum and S. (G.) siamense, whereas heavy dark centromeric bands were present in S. (G.) angulistylum. The best distinguishing character of Simulium species in the subgenus Gomphostilbia is the position of the nucleolar organizer in the short arm of chromosome I. The Ring of Balbiani and the double bubble are located in chromosome arm IIS in all species except for S. (G.) angulistylum, which showed these cytological markers in chromosome arm IIIS. A low chromosomal polymorphism was recorded in all species except for S. (G.) sp. g, which exhibited a standard polytene chromosome. Inversion polymorphisms found in this study conformed to Hardy-Weinberg equilibrium and were not associated with sex. These species have different specific markers and banding patterns although homologous banding sequences were found in chromosome arm IIS in S. (G.) asakoae, S. (G.) sp. g, S. (G.) decuplum and S. (G.) siamense and chromosome arm IIIS in S. (G.) angulistylum. Our results showed no evidence of a sibling species complex within any taxon. PMID- 12733667 TI - Comparative cytogenetics and chromosomal characteristics of ribosomal DNA in the fish genus Vimba (Cyprinidae). AB - Karyotypic and cytogenetic characteristics of Vimba vimba and V. elongata were investigated using differential staining techniques (sequential C-banding, Ag- and CMA3-staining) and fluorescent in situ hybridization (FISH) with 28S rDNA probe. The diploid chromosome number in both species was 2n = 50 with 8 pairs of metacentrics, 14 pairs of submetacentrics to subtelocentrics and 3 pairs of subtelo- to acrocentrics. The largest chromosome pair of the complements was characteristically subtelo- to acrocentric. The nucleolar organizer regions (NORs) in both species were detected in the telomeres of a single, middle-sized subtelocentric chromosome pair, a pattern common in a number of other Leuciscinae. FISH with rDNA probe produced consistently positive hybridization signals detected in the same regions indicated by Ag-staining and CMA3 fluorescence. The distribution of C-positive heterochromatin was identical in both species, including a conspicuous size polymorphism of heterochromatic blocks in the largest metacentric and subtelo- to acrocentric chromosomal pairs. No heteromorphic sex chromosomes were detected. A single analyzed individual of V. melanops possessed the same karyotype and NOR phenotype as V. vimba and V. elongata. The apparent karyotype homogeneity and chromosomal characteristics of ribosomal DNA in all three species of the genus Vimba is consistent to that found in most other representatives of the European leuciscine cyprinid fishes. PMID- 12733668 TI - Genetic differentiation at the 3'UTR of PROS-Ag25: a cDNA homologue of Drosophila melanogaster proteasome subunit PROS-Dm25 in the malaria vector Anopheles gambiae. AB - The Anopheles gambiae cDNA encoding the homologue of the Drosophila melanogaster proteasome PROS-Dm25 was identified and analysed in terms of nucleotide sequence, and chromosomal localisation. In the 3' untranslated region, a GA-rich sequence was mapped which was found to be widely polymorphic among taxa belonging to the A. gambiae complex. PMID- 12733669 TI - Internet-accessible radiographic database of Vietnam War casualties for medical student education. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to determine the feasibility of archiving radiographic images from Vietnam era conflict casualties into a personal computer-based electronic database of text and images and displaying the data using an Internet-accessible database for preservation and educational purposes. MATERIALS AND METHODS: Thirty-two patient cases were selected at random from a pool of 1,000 autopsy reports in which radiographs were available. A total of 74 radiographs from these cases were digitized using a commercial image scanner and then uploaded into an Internet accessible database. The quality of the digitized images was assessed by administering an image-based test to a group of 12 medical students. RESULTS: No statistically significant (p > 0.05) differences were found between test scores when using the original radiographs versus using the digitized radiographs on the Internet-accessible database. CONCLUSION: An Internet-accessible database is capable of effectively archiving Vietnam era casualty radiographs for educational purposes. PMID- 12733670 TI - Gaps in expectations among clients of secondary medical services in the military system compared with the civilian system as a satisfaction index. AB - The Israel Defense Forces Medical Corps operates a health network for Israel Defense Forces soldiers. Secondary medicine is included in the services to which soldiers are entitled. It is provided to military personnel through two parallel systems: within the Medical Corps specialists' clinics and through the auspices of a number of civilian hospital outpatient clinics. The military medical system, like the civilian medical system, is designed to serve its clientele. One of the indices for ascertainment of satisfaction with medical services is compatibility of client expectations with the service actually received. In this study, we present a gap index that demonstrates that there is gap in satisfaction among soldiers receiving secondary medical services from the military network compared with soldiers who receive secondary medical services from the civilian network. We designed a questionnaire administered to 1,532 soldiers and used 1,359 (89% response rate) for our analysis. The military system provides soldiers with services fully in synch with military regulations. Consequently, in most cases, there is a gap between soldiers' expectations from military medical service and the service they receive in practice-a phenomena that impairs soldier satisfaction. On the other hand, soldiers receiving medical services and treatment from the public civilian system receive, for the most part, service and treatment that meets or even exceeds their expectations because the system operates according to other regulations. PMID- 12733671 TI - Skin interface pressure on the NATO litter. AB - The NATO litter serves as a transport device and hospital bed during all types of operations. Little is known about the skin interface pressure on this litter. The purpose of this study was to determine whether various types of padding on the litter and body position affect the peak skin interface pressure and the total body area exposed to interface pressures above 30 mm Hg at different body areas. Thirty-two subjects participated. A repeated measures design was used. The surface effect was statistically significant for all peak pressure and surface area analyses (repeated-measures analysis of variance, p < 0.01). There was a significant decrease in peak pressure and surface area between the litter and litter plus aeromedical evacuation mattress. The addition of the blanket did not significantly reduce pressures and should not be considered a pressure-reducing measure. Conversely, an aeromedical evacuation mattress should be used for all high-risk patients if feasible. Preventive measures (turning, elevating the heels) are still required. PMID- 12733672 TI - Lessons learned from a successful MEDRETE in El Salvador. AB - Medical readiness education and training exercises are short-term exercises designed to provide health care and preventive medicine education to underserved civilian populations overseas. These high profile missions provide superb training opportunities, build democracies, and can be a powerful incentive to retain soldiers in the Reserves. Despite this, the literature offers little guidance in terms of how to best conduct a MEDRETE, particularly with a unit that has not been recently deployed. A U.S. Army Reserve unit was deployed to El Salvador following two devastating earthquakes and treated 20,890 patients in 10 days. This patient volume was achieved by a close cooperative effort among an experienced Mission Coordinator and Reservists and superb host nation support. Lessons learned regarding predeployment, deployment, patient management, and safety issues are presented to assist future units in conducting successful medical readiness education and training exercises. PMID- 12733673 TI - Estimating selected disease and nonbattle injury Echelon I and Echelon II outpatient visits of United States soldiers and Marines in an operational setting from corresponding Echelon III (hospitalizations) admissions in the same theater of operation. AB - Currently, disease and nonbattle injury estimates are based on hospital admission logs and not on battalion aid stations nor medical company patient logs. A significant amount of morbidity can be overlooked and can mislead commanders and their staffs into believing that their unit is at a higher strength level. The objective of this article is to provide a tool for estimating these "hidden" disease and nonbattle injury cases. Nine weeks of military hospitalization data (687 admissions) separated into 14 diagnosis categories were compared with respective outpatient morbidity data (12,109 outpatient visits) during Operation Restore Hope in Somalia in 1993. Proportionate values and 95% confidence intervals were calculated for 5 of the 14 diagnostic categories and total morbidity. These proportionate values were then used to show how disease and nonbattle injury (total and diagnosis category) could be estimated based on hospitalization admissions during a similar military operation. Mission and geographical location information should be considered when using this model, along with other medical planning publications. PMID- 12733674 TI - Exertional heat illness in Air Force basic military trainees. AB - OBJECTIVE: Exertional heat illnesses (heat stroke, heat exhaustion, and heat cramps) are a serious problem in military operations. Air Force basic military training at Lackland Air Force Base is a prime example of where exertional heat illnesses can occur. This project was conducted to evaluate the problem o exertional heat illnesses during basic military training a Lackland Air Force Base. METHODS: Data were obtained from the Ambulatory Data System, the Air Force Reportable Event Surveillance System, and a published report in the December 1999 issue of Military Medicine (Drehner, Neuhauser, Neuhauser, and Blackwood). RESULTS: In 1999, there were 51 cases of heat illness (not necessarily exertional) among basic trainees at Lackland Air Force Base, for a rate of 1.3 per 1,000 trainees. Many of the cases (21) had the nonspecific diagnosis of "heat effect," 4 were diagnosed as heat syncope, 22 as heat exhaustion, and 5 as heat stroke. From 1956 to 1999, there were seven deaths among trainees at Lackland Air Force Base due to heat stroke. CONCLUSION: After reviewing Air Force basic military training, it is apparent that the heat stress incident rates are low. Heat injury rates are low despite being located in an area that has extremely hot temperatures. However, there are some areas, such as tracking and reporting heat illness that need improvement. PMID- 12733675 TI - Description of the oxygen concentration delivered using different combinations of oxygen reservoir volumes and supplemental oxygen flow rates with the Ohmeda Universal Portable Anesthesia Complete draw-over vaporizer system. AB - The Ohmeda Universal Portable Anesthesia Complete system is used in austere conditions where oxygen resources are limited and must be conserved. The purpose of this study was to describe the concentration of oxygen delivered with different combinations of seven oxygen reservoir volumes and four oxygen flow rates. The Medical Education Technologies Incorporated Human Patient Simulator reproduced human physiological tidal volumes of four simulated patients of different weights based upon a typical soldier's weight. Using least squares multiple nonlinear regression, a formula for the curves of the oxygen concentrations was developed. The analysis, across the different patient weights, showed no appreciable increase in oxygen concentration beyond a reservoir volume of 260 mL. Our findings suggest the current standard universal portable anesthesia complete reservoir may not provide optimal oxygen delivery, therefore, we recommend the current reservoir volume be increased from 130 to 260 mL. PMID- 12733676 TI - Rapid diagnosis of an ulnar fracture with portable hand-held ultrasound. AB - Orthopedic fractures are a common injury in operational activities, which are injuries that often occur in isolated or hostile environments. Clinical ultrasound devices have become more user-friendly and lighter, allowing them to be easily transported with forward medical teams. The bone-soft tissue interface has a very large acoustic impedance with a high reflectance that can be used to visualize breaks in contour including fractures. Herein reported is a case of an ulnar fracture that was quickly visualized in the early phase of a multisystem trauma resuscitation with a hand-held ultrasound device. The implications for operational medicine are discussed. PMID- 12733677 TI - Field verification of Test-mate ChE. AB - The objective was to evaluate the ability of the Test-mate ChE to determine acetylcholinesterase (AChE) activity under field conditions. To mimic nerve agent exposure, the U.S. Army Medical Research Institute of Chemical Defense spiked blood samples with variable amounts of soman. Blinded to the identity of the samples, the 520th Theater Army Medical Laboratory tested the samples during a field training exercise inside their environmentally controlled mobile facility. The technicians repeated measurements for 6 consecutive days, and on 1 of the days repeated the measurements six times. The technicians accurately identified all of the samples and quantified the AChE activity. The major trend was that the Test-mate ChE is more precise and reproducible for smaller doses of soman. The results were reliable over all temperatures during the field exercise. In conclusion, the Test-mate ChE is a reliable field instrument to determine blood AChE activity. PMID- 12733678 TI - Interest in healthy diet and physical activity interventions peripartum among female partners of active duty military. AB - Overweight and obesity among soldiers and their dependents have increased over the last decade, mirroring rates in the general population. In general, few programs that result in sustained weight loss have been evaluated, although effective interventions could have clear health and cost benefits for the military. For women, the postpartum period represents a "teachable moment" to promote healthy diet and exercise behaviors related to weight loss, but the attitudes and preferences for weight-loss interventions in this population are unknown. With a view to developing a weight-loss intervention tailored to this population, we surveyed 161 peripartum women at a military base to assess their interests and preferences. Eighty-six percent were dependents. Despite their youth, more than one-third reported entering pregnancy overweight or obese. Interest was high for interventions that promote physical activity and facilitate social interaction. Based on these results, a postpartum exercise intervention is being designed for female partners of active duty soldiers. PMID- 12733679 TI - Oral health disparities between ranks in a military environment: Israel Defense Force as a model. AB - Some studies have demonstrated a relationship between occupational status and position in the workplace to use of dental services and oral health status. Ranks symbolize social status in the military hierarchy, which is different from that accepted in civilian workplace. OBJECTIVE: This study was aimed at analyzing differences between officers and noncommissioned personnel in regard to dental treatment needs and use of dental services. METHODS: One-thousand, one-hundred thirty-nine personnel of the Israel Defense Force were examined, using DMFT (decayed, missing, filled permanent teeth and CPITN (community periodontal index of treatment needs indices to define oral health status. A questionnaire was used to calculate utilization rates. RESULTS: Noncommissioned personnel had on average 50% more unmet caries treatment needs, and 19.1% of them suffered from deep periodontal pockets. No differences were demonstrated in utilization rates CONCLUSIONS: Military rank influences oral treatment needs the way civilian organizational structure does with the exception of utilization rates of dental services. PMID- 12733680 TI - A new model of blast injury from a spherical explosive and its special wound in the maxillofacial region. AB - OBJECTIVE: The goal of this study was to establish an efficient explosive model with spherical explosives for the study of blast injury and its wound characteristics in the maxillofacial region. METHODS: Different spherical explosives were used to produce blast waves and to establish the wound model. Fresh sheep heads as the targets were used for the study of wound characteristics. The wounding parameters and the injuries of targets after explosion were observed. RESULTS: The overpressure values of blast waves decreased exponentially along with the increase of distances to explosives and so did the tissue damage. The blast injury is quite different from other injuries and is more serious. A kind of special "split fracture" of the mandibles was found. CONCLUSIONS: The new explosive wound model with spherical explosives can be used for the study of blast injury in the laboratory. The injury manner of blast wave and the local anatomy of tissues resulted in the special wound characteristics. PMID- 12733681 TI - Frontotemporal dementia affecting a U.S. Air Force officer. AB - We present the case of a 51-year-old former U.S. Air Force officer who developed marked behavioral and personality changes over a 9-year period, ultimately leading to his discharge from the Air Force at a rank below that which he achieved during service. Clinical diagnostic features, neuropsychological testing and neuroimaging together confirmed a diagnosis of frontotemporal dementia. This form of presenile dementia is discussed with reference to his case. PMID- 12733682 TI - The patellofemoral pain syndrome in recruits undergoing military training: a prospective 2-year follow-up study. AB - Patellofemoral pain syndrome is a frequent phenomenon among recruits undergoing military training. The causes, however, are largely unknown, which makes it difficult for medical staff and military personnel to deal with the problem. A prospective 2-year follow-up study was performed in 58 recruits with patellofemoral pain that had started prior to or during basic military training. Anthropometric data and patient history were recorded, and a clinical examination of the knee joint was performed according to established criteria at the time of inclusion into the study. At follow-up investigation, a questionnaire concerning the patient's current physical status and his assessment of his former military seniors were filled out. A large number of patients were engaged in jobs or sports that burdened the knee; previous injuries were also very common. The majority of our patients adhered to the recommended period of reduced activity. However, retrospectively the patients did not feel military personnel had understood them. Less activity appears to have been the main factor that reduced symptoms and complaints. PMID- 12733683 TI - Increasing resident research in a military internal medicine program. AB - The internal medicine residents from our training program rarely participated in scholarly activity prior to 1994. In an attempt to increase the quality and quantity of resident research, we initiated a research program that included: (1) a mandatory research project, (2) 2 months of dedicated research time, (3) appointment of a research director, (4) lectures on the critical appraisal of medical literature and research design, (5) technical support, (6) faculty mentoring, (7) a research meeting and competition involving five residency programs, and (8) military achievement awards. From 1994 to 1999, our house staff has had 134 research presentations and 21 manuscripts accepted for publication in peer-reviewed journals. Ninety percent of our residents presented at least one project at a scientific meeting by completion of their training between 1996 and 1999. Resident scholarly activity is significantly enhanced by a structured research program, an opportunity to present at a scientific meeting, and award recognition. PMID- 12733684 TI - Primary epiploic appendagitis: diagnosis, management, and natural course of the disease. PMID- 12733685 TI - A 36-year-old military recruit with recurrent myalgias and weakness. AB - The objective of this study was to discuss a comprehensive yet cost-effective approach to working up active duty patients with recurrent rhabdomyolysis. A 36 year-old male Army recruit was evaluated at Walter Reed Army Medical Center for recurrent rhabdomyolysis. This case illustrates a practical and cost-effective approach to this goal. Questions along the way are meant to highlight military relevance and expand diagnostic considerations of muscle pain beyond the usual. The case takes an interesting twist and reinforces that, as providers, we must explore every aspect of our patient's history to the fullest to be effective diagnosticians. PMID- 12733686 TI - Canadian perspectives in glaucoma management: setting target intraocular pressure range. AB - There is an abundance of evidence from recent randomized clinical trials showing that lowering the IOP is beneficial to the optic nerve and visual field. Setting and achieving a target IOP range is in keeping with evidence-based medicine. The benefits of reaching this target must be weighed against the risks of the treatment itself. Target IOP is a dynamic concept, needing constant reevaluation. What is lacking are established guidelines for determining the target IOP range that can be used in general ophthalmology practice. PMID- 12733687 TI - Cataract surgery at the Vancouver Eye Care Centre: do patient indications meet provincial clinical practice guidelines? AB - BACKGROUND: Cataract surgery, already the most common elective surgical procedure among patients over the age of 65 in North America, is rising in volume, because the population is aging while at the same time holding growing expectations of continued vitality. In meeting the need, there is a public interest in ensuring that cataract surgery is used appropriately. The purpose of this paper was to investigate the indication for cataract surgery performed in adults at a large tertiary care centre in Vancouver and to determine the extent to which patient criteria met the guidelines of the College of Physicians & Surgeons of British Columbia. METHODS: Patients attending the centre were enrolled in the cataract outcome assessment program through systematic sampling that included every patient with a personal health card number ending in 5 or 8 (20% of patients). There were no patient exclusion criteria. Pre-, intra- and postoperative clinical information was collected from patients' medical records. Patients were also sent a pre- and postoperative visual function and quality-of-life questionnaire. RESULTS: A total of 1098 cataract surgery procedures were performed in 896 patients from March 1999 to December 2000. The average age of the patients was 72.9 years; most (69.5%) were women. Just over half of the procedures (620 [56.6%]) were performed as first-eye surgery. In 191 cases (17.9%) the preoperative visual acuity was 20/40 or better in the affected eye. In most cases (82.1%) visual acuity of 20/50 or worse was the indication for surgery. In 89 cases (8.3%) there was not enough information in the chart to justify the surgery. INTERPRETATION: By examining standard information provided in patients' medical records, we were able to determine the indication for cataract surgery in 92% of cases. The most common indication was poor visual acuity. PMID- 12733688 TI - Microdissection combined with the polymerase chain reaction to identify potentiating viral co-infection in patients with HIV/AIDS with ocular infection. AB - BACKGROUND: In the presence of several coexisting infections, superimposed tissue necrosis or tissue metaplasia, it may be difficult to recognize standard histologic morphology on hematoxylin-eosin slides. Tissue microdissection combined with the polymerase chain reaction (PCR-MD) offers the advantages of high specificity and relative speed. The objective of this study was to describe the use of PCR-MD in identifying potentiating viral co-infection in patients with HIV/AIDS with retinitis and choroiditis. METHODS: Eyes from two patients with HIV/AIDS with several ocular infections were studied by a variety of techniques, including standard histologic examination, immunochemistry, electron microscopy and in situ hybridization. PCR-MD was used to identify coexisting viral infections. RESULTS: Histologic examination showed cytomegalovirus retinitis in both cases. Use of PCR-MD allowed the identification of Epstein-Barr virus within a zone of fulminant varicella-zoster virus retinitis in one patient. PCR-MD confirmed the presence of human herpesvirus 8 in the second patient, who had ocular lymphoma. INTERPRETATION: PCR-MD can be used to demonstrate coexisting viral infection in ocular specimens from patients with unusually fulminant courses. Co-infections may contribute to the observed clinical course and should be considered in patients with rapid progression or unusual presentation. PMID- 12733690 TI - Dysmorphophobia. PMID- 12733689 TI - Ondansetron for the prevention and treatment of nausea and vomiting following pediatric strabismus surgery. AB - BACKGROUND: Neither droperidol nor ondansetron has been proven completely effective, and there are conflicting data comparing the efficacy of the two agents. The purpose of this study was to compare the efficacy, safety and cost of a combination of ondansetron administered intravenously in the operating room followed by oral ondansetron treatment at home with the more commonly used treatment of intravenous droperidol therapy and oral dimenhydrinate therapy, for the prevention and treatment of postoperative nausea and vomiting in children undergoing strabismus surgery. METHODS: Double-blind randomized clinical trial with parallel comparison groups. All patients aged 6 months to 18 years who underwent strabismus surgery at a pediatric hospital in Montreal between Nov. 13, 2000, and June 12, 2001, were included. The exclusion criteria were nausea or vomiting, or use of antiemetics or narcotics in the 24 hours preceding surgery, and past history of hepatic, gastric or renal disease. The outcome measures were frequency of nausea and vomiting, severity of nausea and adverse effects in hospital, during transportation home and during the first 24 hours at home. Data were obtained through nursing notes and through a telephone interview conducted 24 to 48 hours after discharge. RESULTS: Of the 208 eligible patients, 172 were randomly assigned to the study groups (88 to the ondansetron group and 84 to the droperidol/dimenhydrinate group). We found no statistically significant difference in the incidence of nausea and vomiting in hospital or at home between the two groups (25.3% vs. 31.6%, p = 0.371). There was a significant difference between the two groups in the rate of vomiting during transportation home (3.6% vs. 12.6%, p = 0.044). The incidence of severe nausea was 14.4% with ondansetron and 15.4% with droperidol, a nonsignificant difference (p = 1.00). No significant difference was observed between the two groups in the incidence of any nausea (p = 0.434) or adverse effects (p = 0.220). We calculated that the combination of droperidol and dimenhydrinate was seven times less costly than the ondansetron regimen. INTERPRETATION: In this study, the efficacy and safety of intravenous administration of droperidol followed by oral use of dimenhydrinate did not differ from that of intravenous followed by oral use of ondansetron in children undergoing strabismus surgery. Since treatment with ondansetron is much more costly than the combination of droperidol and dimenhydrinate, at this time the use of ondansetron in the prevention and treatment of vomiting and nausea in this population may not be beneficial on a cost basis if all other variables are considered. PMID- 12733691 TI - Rhodotorula infection in a corneal graft following penetrating keratoplasty. PMID- 12733692 TI - Primary choroidal melanoma masquerading as a hemorrhagic lesion in a patient with ocular trauma. PMID- 12733693 TI - Acute angle-closure glaucoma following neodymium:YAG laser posterior capsulotomy. PMID- 12733694 TI - Effects of cholesterol-lowering therapy on pressor hyperreactivity to stress in hypercholesterolemic patients. AB - We previously demonstrated that normotensive patients with hypercholesterolemia showed excessive blood pressure (BP) responses to stress tests. In this study, we examined the effects of cholesterol-lowering therapy on BP in order to confirm that hypercholesterolemia plays a role in the regulation of BP. Fifteen patients with hypercholesterolemia and 24 normal cholesterolemic controls performed mental arithmetic stress (AS) and hand grip (HG) tests. BP was measured during the tests. Serum lipids and lipid peroxides were measured before the AS. Platelet intracellular free calcium concentration ([Ca2+])i with and without low density lipoprotein (LDL) stimulation, plasma cGMP and NOx were determined immediately before AS and at the end of each test. In hypercholesterolemic patients, the tests were repeated at the end of a 12-week treatment with 10 mg/day of pravastatin, a hepatic hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase inhibitor. In hypercholesterolemic patients, BP responses to both tests were greater than those of the controls. Basal and LDL-stimulated platelet [Ca2+]i were higher, and the ratio of plasma cGMP to NOx was lower. Serum total cholesterol, LDL cholesterol and lipid peroxides were significantly decreased in association with the pravastatin treatment. Systolic BP to AS and systolic BP/diastolic BP to HG were decreased (p < 0.01, p < 0.01/p < 0.02, respectively). Platelet [Ca2+]i with LDL stimulation was decreased (p < 0.01). Plasma cGMP was increased (p < 0.05), whereas NOx was decreased (p < 0.05); therefore, the ratio of cGMP to NOx was increased (p < 0.05). In conclusion, excessive blood pressure responses to stress tests were improved after cholesterol-lowering therapy. This finding suggests that hypercholesterolemia itself is involved in the regulation of BP. PMID- 12733695 TI - Relationship among systolic blood pressure, serum insulin and leptin, and visceral fat accumulation in obese children. AB - The aim of this study was to clarify the relation among systolic blood pressure (SBP), serum insulin, leptin, visceral fat accumulation and family history of hypertension, and to elucidate the pathophysiologic mechanism of blood pressure elevation in obese children. This study examined 109 obese children with a family history of hypertension (OF: 77 boys and 32 girls), and 83 obese children without such a history (ON: 60 boys and 23 girls). Body height and weight, and percent of body fat were measured and the percent of relative weight was calculated. Both boys and girls, the two groups were matched with respect to age, height, and weight. SBP was measured in the seated position using an automated recorder. Abdominal fat thickness (maximum preperitoneal fat thickness: Pmax; minimum subcutaneous fat thickness: Smin) were measured using ultrasonography. The fasting serum levels of insulin and leptin were measured by radioimmunoassay. All subjects were simply obese, without diabetic states. In both OF and ON, SBP was associated with insulin levels, leptin levels, and Pmax by simple regression analysis, and with insulin levels by stepwise regression analysis. Insulin levels were associated with leptin levels and Pmax by simple regression analysis, and with leptin levels by stepwise regression analysis. These findings indicated that SBP was associated with hyperinsulinemia, hyperleptinemia and visceral accumulation regardless of a family history of hypertension in obese children, as well as later in adult obesity. For primary prevention of hypertension, these results support the importance of implementation of a strategy to prevent obesity, especially visceral obesity. An effective strategy for preventing childhood obesity will contribute to a future decrement in cases of metabolic syndrome, including adulthood hypertension. PMID- 12733696 TI - Estimated protein intake and blood pressure in a screened cohort in Okinawa, Japan. AB - We examined the relation between protein intake and blood pressure in a screened cohort in Okinawa, Japan. A total of 1,299 screened subjects, 885 men and 414 women, were examined at the Okinawa General Health Maintenance Association. Daily intake of sodium (Na) and potassium (K) was estimated from Na, K, and creatinine excretion by the method of Kawasaki et al., and daily protein intake was estimated by the method of Maroni et al. as the estimated daily urinary excretion of urea nitrogen. Mean (SD) daily protein intake was 71.8 (18.6) g in men and 54.0 (13.5) g in women, and the mean (SD) daily protein intake per unit kg body weight was 1.1 (0.2) g/kg in men and 1.0 (0.2) g/kg in women. In men, both systolic blood pressure (SBP) and diastolic blood pressure (DBP) were higher in those with lower protein intake (LP; < 1.0 g/kg/day) than in those with higher protein intake (HP; > or = 1.0 g/kg/day) (p < 0 .05 for DBP). In women, both SBP and DBP were higher in those with LP than in those with HP, but these differences were not statistically significant. However, urinary excretion of both Na and K was lower in those with LP than in those with HP, respectively, both in men and women (p < 0.0001). In summary, estimated daily protein intake was about 1.1 g/kg in men and 1.0 g/kg in women. Despite the higher urinary excretion of Na, both SBP and DBP tended to be lower in those with higher daily protein intake, particularly in men. PMID- 12733697 TI - Influences of increased oxidative stress on endothelial function, platelets function, and fibrinolysis in hypertension associated with glucose intolerance. AB - The effect of oxidative stress on endothelial function, platelet function, and fibrinolysis in hypertension with or without glucose intolerance was examined. The endothelium, platelets and fibrinolysis play important roles in the progression of atherosclerosis and interact with each other. We have previously demonstrated that glucose intolerance impairs endothelial function in hypertension, but its precise mechanisms have not been clarified. Hypertensive patients were divided by the results of 75-g oral glucose tolerance test into a normal glucose metabolism group (n = 65) and a glucose intolerance group (n = 47). The plasma level of thiobarbituric acid-reactive substances (TBARS) was assessed as a marker of oxidative stress. Endothelial function was assessed by flow-mediated dilatation (FMD), platelet function by the concentration of ADP dose inducing half-maximal aggregation (EC50), and fibrinolytic parameters by radioimmunoassay. These functions were assessed before and after acute administration of vitamin C. FMD was reduced while TBARS and fibrinolytic parameters were higher in patients with glucose intolerance than in those with a normal glucose metabolism. Vitamin C increased FMD and reduced fibrinolytic parameters significantly in the glucose intolerance group, but not in the group with normal glucose metabolism. On the other hand, the EC50 was similar in both groups. In conclusion, glucose intolerance aggravates oxidative stress, thereby contributing to the impairment of endothelial function in patients with hypertension. These abnormalities affect fibrinolysis but not platelet function. PMID- 12733699 TI - Bezafibrate improves hypertension and insulin sensitivity in humans. AB - We examined cellular membrane fatty acid composition and insulin sensitivity in patients with mild essential hypertension and hyperlipidemia, and investigated whether bezafibrate, a lipid-lowering drug, could improve elevated blood pressure and insulin sensitivity in these subjects by ameliorating cellular membrane fatty acid composition. Twenty-seven subjects were recruited. Twelve men with mild essential hypertension [systolic blood pressure (SBP) between 140 mmHg and 160 mmHg] and hypertriglyceridemia (plasma triglyceride concentration over 150 mg/dl) were designated the HL group. Fifteen men with mild essential hypertension and normotriglyceridemia (plasma triglyceride concentration below 150 mg/dl) were designated the NL group. Subjects in the HL group were given bezafibrate 400 mg/dl and those in the NL group were given placebo for 3 months. Bezafibrate significantly reduced SBP (140 +/- 2.6 to 131.8 +/- 2.6 mmHg, mean +/- SEM), diastolic blood pressure (DBP) (87.8 +/- 2.0 to 82.8 +/- 2.6 mmHg), fasting plasma triglyceride concentration (225.5 +/- 23.5 to 102.9 +/- 10.9 mg/dl), fasting plasma insulin concentration (9.6 +/- 0.8 to 7.1 +/- 0.8 microU/ml), and homeostasis model assessment scores (HOMA-R, 2.4 +/- 0.2 to 1.7 +/- 0.2), and significantly improved the insulin sensitivity index (56.0 +/- 3.0 to 70.7 +/- 4.8 mg x l2/mmol x mU x min) in the HL group. Regarding erythrocyte membrane fatty acid composition, bezafibrate reduced the percentages of saturated fatty acids (SFA) and increased the percentage of polyunsaturated fatty acids (PUFA). Plasma triglyceride concentrations were positively correlated with HOMA-R (r = 0.50, p < 0.01) and SFA (r = 0.39, p < 0.05), and negatively correlated with PUFA (r = -0.45, p < 0.05) before administration of placebo or bezafibrate. In conclusion, an improvement of hyperlipidemia by bezafibrate may be attributed to reduction of blood pressure and amelioration of insulin sensitivity. Abnormalities in membrane lipid composition may play an important role in these metabolic disorders. PMID- 12733698 TI - An alternative fast and convenient genotyping method for the screening of angiotensin converting enzyme gene polymorphisms. AB - Insertion/deletion (I/D) polymorphisms in intron 16 of the angiotensin converting enzyme gene (ACE) are associated with the plasma angiotensin converting enzyme (ACE) levels, and individuals with the DD allele have been reported to be more susceptible to cardiovascular disease than those without. The conventional genotyping method for the screening of I/D polymorphisms, which involves polymerase chain reaction (PCR)-gel electrophoresis, is laborious and time consuming. In this study, we assessed the use of TaqMan-PCR genotyping for the screening of I/D polymorphisms as a replacement for the conventional method. We genotyped seven single nucleotide polymorphisms (SNPs) in linkage disequilibrium (LD) with the I/D polymorphisms, and calculated the LD coefficients of the I/D polymorphisms. We found that three polymorphisms, rs4331, rs4334 and rs4341, exhibited the highest LD coefficients (D' = 1.000; r2 = 0.967) and that the genotyping of rs4341 by the TaqMan-PCR method yielded the best discrimination among the different genotypes. Genotyping of 511 samples took only 2 h and the amount of DNA required for each test was only 6 ng by the TaqMan-PCR method using rs4341. In the course of this study, we identified a novel additional polymorphism (a deletion of six amino acids) in exon 13, near rs4316. The deletion allele encoded the testicular ACE, but not the plasma ACE. We concluded that genotyping of the rs4341 ACE polymorphism by the TaqMan-PCR method is a fast and convenient alternative method for direct I/D genotyping. We also concluded that testicular ACE may manifest a deletion of six amino acids that may result in deleterious function of this enzyme. PMID- 12733700 TI - The renin-angiotensin system is involved in the production of plasminogen activator inhibitor type 1 by cultured endothelial cells in response to chylomicron remnants. AB - Triglyceride-rich lipoproteins have been suggested to promote atherosclerosis. Plasminogen activator inhibitor type 1 (PAI-1) plays an important role in the events of cardiovascular pathophysiology. The renin-angiotensin system influences various vascular functions, including PAI-1 production. We examined whether or not chylomicron remnants increased PAI-1 mRNA and protein production in endothelial cells and whether or not an inhibition of the renin-angiotensin system interfered with this effect. Chylomicron remnants were isolated from functionally hepatectomized rats injected with chylomicrons. Human umbilical vein endothelial cell cultures (HUVECs) were incubated with chylomicron remnants with or without an angiotensin-converting enzyme inhibitor (temocaprilat), an angiotensin II receptor type 1 antagonist (RNH-6270), or an angiotensin II receptor type 2 antagonist (PD123319). Chylomicron remnants increased PAI-1 secretion in HUVECs (0.5 microg/ml; 128.3 +/- 6.1%, the mean +/- SEM) as well as angiotensin II (10 nmol/l; 130.7 +/- 9.5%) in 18 h, as compared with the controls, as well as stimulated PAI-1 mRNA expression to a maximum level at 4 h. Temocaprilat and RNH-6270, but not PD123319, attenuated all of these effects. Chylomicron remnants enhanced nuclear extract binding to a very low-density lipoprotein response element in the PAI-1 promoter region and activated nuclear factor-kappaB. Extracellular signal-regulated kinase (ERK 1/2) was phosphorylated in response to chylomicron remnants. These effects were inhibited by temocaprilat or RNH-6270. In conclusion, chylomicron remnants increased protein secretion and mRNA expression of PAI-1 in HUVECs. Inhibition of the renin-angiotensin system reduced this stimulation. PMID- 12733701 TI - Enhanced depressor response to endothelial nitric oxide synthase gene transfer into the nucleus tractus solitarii of spontaneously hypertensive rats. AB - Previously, we demonstrated that endothelial nitric oxide synthase (eNOS) gene transfer into the nucleus tractus solitarii (NTS) decreased blood pressure, heart rate and sympathetic nerve activity in conscious normotensive Wistar-Kyoto rats (WKY). In order to determine whether overexpression of eNOS in the NTS causes different effects on blood pressure and heart rate between spontaneously hypertensive rats (SHR) and WKY, we transfected adenovirus vectors encoding either eNOS (AdeNOS) or beta-galactosidase (Ad beta gal) into the NTS of SHR and WKY in vivo. The local expression of eNOS in the NTS was confirmed by Western blot analysis for eNOS protein, and the magnitude of expression did not differ between SHR and WKY. Blood pressure and heart rate were monitored by the use of a radio-telemetry system in a conscious state before and 7 days after the gene transfer. Systolic blood pressure (SBP) and heart rate decreased on day 7 in both AdeNOS-transfected SHR and WKY. However, the magnitude of decreases in SBP of AdeNOS-transfected SHR was greater than that of AdeNOS-transfected WKY (-24.1 +/- 2.9 vs. -15.9 +/- 2.1 mmHg, p < 0.05). Transfection of Ad beta gal into the NTS did not alter SBP in either group. A depressor response evoked by microinjection of L-glutamate into the NTS did not differ between the two strains. These results suggest that overexpression of eNOS in the NTS causes a greater depressor response in SHR than in WKY in a conscious state. An abnormality of the L arginine-NO pathway in the NTS may be related to the hypertensive mechanism(s) of SHR. PMID- 12733702 TI - Sexual dimorphism of 11beta-hydroxysteroid dehydrogenase in hypertensive and normotensive rats. AB - To evaluate the role of sexually dimorphic tissue expression of 11beta-oxidase activity of 11beta-hydroxysteroid dehydrogenase (11betaHSD) in gender-associated blood pressure differences, we have studied female and male hypertensive rats of two different strains and their normotensive controls: spontaneously hypertensive rats (SHR), Wistar-Kyoto rats (WKY) and Dahl salt-sensitive (SS/Jr) and salt resistant rats (SR/Jr). In hypertensive SHR and SS/Jr, but not in normotensive strains WKY and SR/Jr, blood pressure reached a higher level in males than in females. The activity of 11betaHSD was higher in the renal cortex, medulla, colon and aorta of males than of females in all investigated strains with the exception of aortic 11betaHSD in SHR and WKY rats, both of which had very low 11beta oxidase activity. In contrast to gender-dependent differences, strain differences of 11betaHSD were observed in a limited number of tissues only. Renal medullary 11betaHSD showed significantly lower activity in WKY than in SHR, whereas no difference was observed in the renal cortex. Similarly, colonic 11betaHSD activity was lower in WKY than in SHR. In Dahl rats the strain differences were observed in aortic 11betaHSD that had higher activity in SR/Jr than in SS/Jr rats; no difference was observed in the kidney or colon. These data demonstrate the following. 1) Sexual dimorphism of 11betaHSD activity exists in the kidney, colon, and aorta. 2) The sexual dimorphism of 11betaHSD does not play a role in gender-associated differences in blood pressure. 3) The reduced 11betaHSD activity in the aorta of hypertensive SS/Jr compared to SR/Jr rats suggests that this enzyme might play a role in the pathogenesis of salt-sensitive hypertension in Dahl rats. PMID- 12733703 TI - Hypertension accelerates diabetic nephropathy in Wistar fatty rats, a model of type 2 diabetes mellitus, via mitogen-activated protein kinase cascades and transforming growth factor-beta1. AB - Although it is known that diabetic nephropathy is accelerated by hypertension, the mechanisms involved in this process are not clear. In this study we aimed to clarify these mechanisms using male Wistar fatty rats (WFR) as a type 2 diabetic model and male Wistar lean rats (WLR) as a control. Each group was fed a normal or high sodium diet from the age of 6 to 14 weeks. We determined the blood pressure and urinary albumin excretion (UAE). At the end of the study, the expressions of mitogen-activated protein kinases (MAPK) and transforming growth factor-beta1 (TGF-beta1) were examined in the isolated glomeruli by Western blot analysis, and the number of glomerular lesions was determined by conventional histology. High sodium load caused hypertension and a marked increase in UAE in the WFR but not in the WLR. Glomerular volume was increased in the hypertensive WFR. There was no difference among the four groups in the expression of c-Jun-NH2 terminal kinase (JNK). In contrast, the expressions of extracellular signal regulated kinase 1/2 (ERK1/2) and its upstream regulator, MAPK/ERK kinase 1 (MEK1), were augmented in the hypertensive WFR. Expression of p38 MAPK was increased in the normotensive WFR, and further enhanced in the hypertensive WFR. Moreover, administration of high sodium load to WFR augmented the expression of TGF-beta1. In conclusion, systemic hypertension in WFR accelerates the diabetic nephropathy in type 2 diabetes via MEK-ERK and p38 MAPK cascades. TGF-beta1 is also involved in this mechanism. PMID- 12733704 TI - Osteoporosis is more prevalent in adrenal than in pituitary Cushing's syndrome. AB - Osteoporosis is the most common complication of Cushing's syndrome. We retrospectively examined the prevalence and risk factors for osteoporosis in 42 female patients with Cushing's syndrome. Osteoporosis and atraumatic fractures were assessed by bone mineral density of the lumbar vertebral spine (L2-L4) using dual energy X-ray absorptiometry (DXA) and X-ray examination. The prevalence of osteoporosis and fracture were 54.8% and 21.4%, respectively. The prevalence of osteoporosis (69.6% vs. 37.8%) and atraumatic bone fracture (26.1% vs. 15.8%) were significantly higher in patients with adrenal Cushing's than in those with pituitary Cushing's. AP and lateral BMD was significantly higher in patients with pituitary origin than in those with adrenal origin. Among several variables examined by multiple logistic regression, the etiology of Cushing's syndrome (adrenal vs. pituitary origin) was a significant factor affecting the prevalence of osteoporosis. Neither age, body mass index, duration of amenorrhea, nor extent of hypercortisolism were significant factors in this analysis. Plasma DHEA-S and urinary 17-KS excretion were significantly higher in pituitary Cushing's than in adrenal Cushing's. The present study shows that the prevalence of osteoporosis in patients with Cushing's syndrome is influenced by its etiology. A factor associated with pituitary Cushing's syndrome, such as adrenal androgen, may protect these patients from glucocorticoid-induced osteoporosis. PMID- 12733705 TI - Inhibitory effect of the alpha1-antitrypsin Pittsburgh type-mutant (alpha1-PIM/R) on proinsulin processing in the regulated secretory pathway of the pancreatic beta-cell line MIN6. AB - To elucidate its effect on proinsulin processing, we introduced the expression of a Pittsburgh type-mutant, alpha1-protease inhibitor M/R (alpha1-PIM/R) and its chimera protein with growth hormone (GH) (GHalpha1-PIM/R) into MIN6 cells. In metabolic labeling and chasing experiments with [3H]-Leu and [35S]-Met, proinsulin appeared in the medium during stimulatory secretion only from MIN6 clones expressing GHalpha1-PIM/R and, surprisingly, alpha1-PIM/R, but not from the clones of either the control or alpha1-PI. The major part of alpha1-PIM/R was secreted through the constitutive pathway and about 10% of total secreted alpha1 PIM/R in the chase periods entered the regulated pathway. On the other hand, GHalpha1-PIM/R was mainly transported to the secretory granules and about 80% of the total secreted GHalpha1-PIM/R in the chase periods was secreted during stimulatory secretion. In the first 3 h chase periods without stimulation, only alpha1-PIM/R and no GHalpha1-PIM/R appeared in the medium, thus suggesting that alpha1-PIM/R might be transported through a constitutive-like pathway for those periods. The alpha1-PI, which had no inhibitory effect on proinsulin processing, showed similar secretion pathways to those of alpha1-PIM/R. This implies that some part of alpha1-PIM/R and alpha1-PI entered the regulated pathway, not due to any specific interaction between the processing endoproteases and serine protease inhibitors, but due to some type of passive transport in a nonselective manner. The inhibitory effect of alpha1-PIM/R in the regulated secretory pathway was slightly but clearly evident when it was expressed in MIN6 beta-cells. PMID- 12733706 TI - Regulation of corticotropin-releasing factor (CRF) type-1 receptor gene expression by CRF in the hypothalamus. AB - We reported previously that acute stress and intracerebroventricular (i.c.v.) injection of corticotropin-releasing factor (CRF) increased neuronal activation and CRF type-1 receptor (CRFR-1) mRNA expression in the CRF-producing neurons of the parvocellular paraventricular nucleus (PVN) of the hypothalamus. In this study, to determine whether CRF can act directly on hypothalamic CRF neurons, thereby increasing CRFR-1 expression, microinjection of CRF into PVN neurons in vivo and primary cultures of dispersed rat fetal hypothalami in vitro were performed. Microinjection of 0.1 microg of CRF into the PVN significantly increased c-fos and CRFR-1 mRNA expression in the CRF-producing parvocellular PVN, 30 min or 180 min after injection, respectively. This effect was blocked by a CRF antagonist, alpha-helical CRF. CRF, when injected into the lateral ventricle at the same dose, increased neither CRFR-1 nor c-fos mRNA levels in the PVN. Primary culture of hypothalamic neurons revealed that CRFR-1 like immunoreactivity was located in CRF-containing neurons, and that the CRFR-1 mRNA level was significantly increased 4 h after incubation with 10(-8) M CRF. These results demonstrate that CRF directly affects hypothalamic neurons to increase CRFR-1 mRNA expression, providing evidence of a direct role for CRF in the regulation of CRFR-1 expression of hypothalamic neurons. PMID- 12733707 TI - Pineal gland (melatonin) affects the parturition time, but not luteal function and fetal growth, in pregnant rats. AB - The purpose of the present study was to investigate the role of pineal gland (melatonin) on parturition time, luteal function, and fetal growth in pregnant rats. Cycling rats were subjected to pinealectomy or sham operation under ether anesthesia; and pinealectomized rats immediately underwent implantation of a melatonin capsule (PINX + Mel group) or a vehicle-containing capsule (PINX group), and sham operated rats also underwent implantation of a vehicle containing capsule (control group). All rats were maintained under the same photoperiod conditions (14 L:10 D) and were induced pregnancy. Blood samples were obtained on days 7, 12, 15, 17, 19, and 21 of pregnancy to measure serum progesterone concentrations, and parturition times were recorded on days 22 and 23. In the next experiment, pregnant PINX rats received subcutaneous injection of melatonin (10 microg/body) at 08:00 h (PINX + 8 h group) or at 20:00 h (PINX + 20 h group) from day 15 to the end of pregnancy, and parturition times were recorded. Parturition times of rats in the PINX group, the PINX + Mel group or the PINX + 8 h group, but not the PINX + 20 h group, were significantly different compared with those in the control group. Pinealectomy or melatonin implantation did not affect serum progesterone concentrations during pregnancy or the number and weight of fetuses or corpora lutea. The present results indicate that pineal gland (melatonin rhythm) synchronizing with photoperiodic rhythm is likely to be an important determinant of parturition time, but it does not affect progesterone production or fetal growth in pregnant rats. PMID- 12733708 TI - Practical treatment with minimum maintenance dose of anti-thyroid drugs for prediction of remission in Graves' disease. AB - Although many researchers have reported clinical and laboratory parameters for prediction of remission in Graves' disease during or after anti-thyroid drug therapy, there is no reliable one to assure the complete remission. We prospectively examined a practical therapy with minimum maintenance dose of anti thyroid drugs for prediction of remission in Graves' disease. Fifty-seven patients with Graves' disease were treated with anti-thyroid drugs at the initial dose of 30 mg/day of methimazole (MMI) or 300 mg/day of propylthiouracil (PTU). Then, doses were gradually decreased, and finally discontinued when the patients were able to maintain euthyroid (normal FT4 and TSH) for at least 6 months with the minimum maintenance dose (MMI 5 mg every other day or PTU 50 mg every other day). After discontinuation of drugs, FT4, FT3, TSH and TSH-binding inhibitory immunoglobulin (TBII) were measured every one to two months for the first 6 months and every 3-4 months for the next 18 months to confirm continuous remission. After 2 years of drug cessation, 46 (81%) of 57 patients were in remission and the other 11 patients had relapsed into thyrotoxicosis. At the time of drug discontinuation, the serum concentration of FT4, FT3 and TSH, titers of anti-thyroglobulin antibodies and anti-thyroid microsomal antibodies, goiter size were not different between the remission and relapse groups. At the time of drug cessation, the activities of TBII and thyroid-stimulating antibodies (TSAb) overlapped between the two groups, although they were significantly lower in the remission group than in the relapse group (p<0.01). Forty percent (4/10) of TBII positive patients and 71% (23/32) of TSAb positive patients continued to be in remission. On the other hand, thyrotoxicosis relapsed in 5 (11%) of 47 TBII negative and 2 (8%) of 25 TSAb negative patients. These data indicate that minimum maintenance therapy to keep euthyroid (normal FT4 and TSH) for 6 months is a practical measure for 81% prediction of remission in Graves' disease. The measurement of TBII or TSAb gave little additional information for predicting remission. PMID- 12733709 TI - Effects of hormone replacement therapy on circulating docosahexaenoic acid and eicosapentaenoic acid levels in postmenopausal women. AB - Hormone replacement therapy (HRT) has antiatherosclerotic effects of which the mechanism remains unclear. The ingestion of fish oil or other sources of n-3 polyunsaturated fatty acids has been included in comprehensive strategies to prevent atherosclerosis. Many epidemiologic studies have shown that the dietary intake of docosahexaenoic acid and eicosapentaenoic acid has antiatherosclerotic effects. We investigated the effect of HRT on plasma docosahexaenoic acid and eicosapentaenoic acid concentrations in postmenopausal women. Fifty-nine postmenopausal women, who received conjugated estrogens (0.625 mg/day) and medroxyprogesterone (2.5 mg/day) for 12 months, and 45 control postmenopausal women, who did not receive HRT, volunteered to participate in this study. Plasma docosahexaenoic acid and eicosapentaenoic acid concentrations were measured at baseline and at 6 and 12 months after the start of HRT. HRT significantly increased the plasma docosahexaenoic acid and eicosapentaenoic acid concentrations from 134 +/- 5 microg/ml and 69 +/- 4 microg/ml at baseline to 156 +/- 7 microg/ml and 85 +/- 7 microg/ml after 12 months (both p<0.01). However, the control group showed no significant change in their plasma docosahexaenoic acid and eicosapentaenoic acid levels during the study. HRT increased plasma docosahexaenoic acid and eicosapentaenoic acid levels in postmenopausal women. We propose that the increase in docosahexaenoic acid and eicosapentaenoic acid may be partially responsible for the beneficial mechanisms by which HRT induces an antiatherosclerotic effect in postmenopausal women. PMID- 12733711 TI - The comparison of the effects of short-term growth hormone treatment in patients with achondroplasia and with hypochondroplasia. AB - The effects of recombinant human growth hormone (rhGH) treatment for three years were compared in patients with achondroplasia (ACH) and hypochondroplasia (HCH), whose diagnosis had been confirmed by DNA analysis of the fibroblast growth factor receptor 3 gene. Height SDS (H-SDS) and height velocity SDS (HV-SDS) using the standard for ACH significantly improved during three-year treatment as compared with that before treatment in both ACH and HCH except HV-SDS in the third year. The improvement was much greater in HCH than in ACH. The mean increase H-SDS using the standard for ACH in three years in ACH (from -0.2 SD to 0.1 SD) is almost negligible but that in HCH (from 1.2 SD to 2.6 SD) can be estimated as effective clinically. It can be concluded short-term GH treatment in HCH is effective to increase growth rate and H-SDS, but it has little effect in ACH. Further studies would be required to confirm the other beneficial effects of GH treatment such as increase in bone mineral density in ACH and HCH and the effect on the final height. PMID- 12733710 TI - Increased urinary levels of pentosidine, pyrraline and acrolein adduct in type 2 diabetes. AB - This study investigates whether urinary levels of pentosidine, pyrraline and acrolein adduct are increased in type 2 diabetes (DM), and whether these levels are correlated with glycemic control and clinical traits. Urinary levels of pentosidine, pyrraline and acrolein adduct in DM patients (n = 100) recruited from the outpatient clinic of our university hospital were compared with those of age- and sex-matched non-diabetic subjects (n = 50). The correlation of these urinary levels with the glycemic control and the clinical traits were examined. Furthermore, the influence of smoking habit on the levels of acrolein adduct was examined. Urinary levels of pentosidine, pyrraline and acrolein adduct were all significantly (p<0.001) higher in the DM group than in the non-DM group (pentosidine (log(pmol/mgCr)), 1.579 +/- 0.147 vs 1.427 +/- 0.142; pyrraline (log(nmol/mgCr)), 0.888 +/- 0.402 vs 0.581 +/- 0.336; acrolein adduct (log(nmol/mgCr)), 2.316 +/- 0.221 vs 2.051 +/- 0.201). Glycemic control parameters, such as fasting plasma glucose (FPG) and HbA1c, were significantly correlated with these urinary levels. Age was correlated with the urinary levels of pentosidine but not with those of pyrraline and acrolein adduct. The urinary albumin excretion rate did not correlate with any of these urinary levels. The levels of acrolein adduct were higher in the subjects with smoking habit than in those without the habit in the DM group as well as in the non-DM group (DM, 2.391 +/- 0.230 and 2.212 +/- 0.190, p=0.0004; Non-DM, 2.120 +/- 0.171 and 1.993 +/- 0.206, p=0.0503). The urinary levels of pentosidine, pyrraline and acrolein adduct were increased in DM and were significantly correlated with glycemic control levels. In addition, smoking habit seems to increase the urinary levels of acrolein adduct. PMID- 12733712 TI - Increased expression of phosphorylated p70S6 kinase and Akt in papillary thyroid cancer tissues. AB - Although a number of abnormalities in oncogenes have been reported in thyroid neoplasms, little information is available on the signal transduction pathway involved in neoplastic thyroid cell growth. Both p70S6 kinase (p70S6K) and Akt are kinases downstream of phosphatidylinositol 3 kinase (PI3K). These kinases are phosphorylated and activated by growth factors including IGF-1, EGF/TGF-alpha, and HGF in thyroid cells. Since the receptors for these growth factors are reportedly overexpressed in human thyroid cancer, we hypothesized that the PI3K mediated signalings are overactivated in thyroid cancers. Tumorous and adjacent normal tissues of 20 patients with papillary thyroid cancer were obtained at surgery, and expression of p70S6K and Akt were measured by Western blot. Expression of the protein levels of p70S6K was increased in tumor tissues (T) compared to normal thyroid tissues (N), and expression of phosphorylated p70S6K was also significantly increased in tumor than in surrounding normal tissues. Overexpression of p70S6K in tumor tissues was further confirmed by immunohistochemistry. Strong immunoreactivity in the cytoplasm of thyroid cancer cells was seen in the majority of cases, whereas little immunoreactivity was found in the surrounding normal portion. Expression of phosphorylated Akt (pAkt) was also significantly higher in tumor tissues. Phosphorylation of Bad (pBad), a substrate of Akt, was also increased in the tumor tissues in association with activation of Akt, and the T/N ratio for pAkt positively correlated to the T/N ratio for pBad. The data presented here demonstrate that both p70S6K and Akt are activated in the majority of human papillary cancer cells. Activation of these signalings may be involved in the progression of papillary carcinoma by stimulating cell proliferation and/or preventing apoptosis. PMID- 12733713 TI - No evidence of radiation risk for thyroid gland among schoolchildren around Semipalatinsk Nuclear Testing Site. AB - To assess thyroid status among the schoolchildren around Semipalatinsk Nuclear Testing Site (SNTS), Kazakhstan, and to evaluate the current status of iodine deficiency in this area, we performed medical screening of schoolchildren in two villages, Kaynar and Karaul villages, East Kazakhstan Region, Republic of Kazakhstan, located within 100 km of SNTS. A total of 196 schoolchildren were chosen at random. Control groups comprised 250 schoolchildren from Nagasaki, an iodine-rich area, and 100 schoolchildren from Gomel, an iodine-deficient area contaminated by the Chernobyl Nuclear Power Plant accident. Ultrasound screening of thyroid revealed three cases of benign thyroid disease (two cases of goiter and one single cyst), but no cases suspicious of malignancy. The urinary iodine (UI) concentrations of subjects in Kaynar and Karaul ranged from 21.8 to 735.8 microg/L, 4.3% of whom showed low UI concentrations (<50 microg/L), compared with 0% in the Nagasaki group and 52% in the Gomel group. The median UI concentration in Kaynar and Karaul was 153.2 microg/L, which was significantly lower than that in Nagasaki (366.3 microg/L, p<0.0001) but higher than that in Gomel (47.3 microg/L, p<0.0001). In conclusion, there was a low incidence of morphological abnormalities in the thyroid, and no evidence for severe iodine deficiency among the Kazakhstani children studied. These results suggest that there is no transgenerational risk for schoolchildren born from parents irradiated as a result of tests carried out in SNTS. PMID- 12733714 TI - A family of autosomal dominant hypocalcemia with an activating mutation of calcium-sensing receptor gene. AB - Autosomal dominant hypocalcemia (ADH) caused by activating mutations of calcium sensing receptor (CaSR) is characterized by hypocalcemia with inappropriately low concentration of PTH and relative hypercalciuria. Active vitamin D treatment often leads to nephrolithiasis and renal impairment in patients with ADH. However, differential diagnosis between ADH and idiopathic hypoparathyroidism is sometimes very difficult. Here, we report a mutation of CaSR and its functional property found in three generations of a Japanese family. The proband developed seizures at 7 days of age. His mother and elder sister were discovered to have hypoparathyroidism by family survey, but his father was normocalcemic. His grandfather developed heart failure and was found to have hypoparathyroidism. All affected members had been treated with active vitamin D3 and bilateral nephrolithiasis were detected in three of them. DNA sequencing revealed that all affected patients had a heterozygous mutation in CaSR gene that causes proline to leucine substitution at codon 221 (P221L). In vitro functional analysis of the mutant CaSR by measuring inositol 1,4,5-trisphosphate production in response to changes of extracellular Ca indicated that this mutation is an activating one and responsible for ADH in this family. Therefore, careful monitoring of urinary Ca excretion before and during treatment of PTH-deficient hypoparathyroidism is very important, and screening of CaSR mutation should be considered in patients with relative hypercalciuria or with a family history of hypocalcemia. PMID- 12733716 TI - Expression of 20alpha-hydroxysteroid dehydrogenase mRNA in human endometrium and decidua. AB - Progesterone secreted from ovarian corpus luteum plays pivotal roles in endometrial differentiation, and local progesterone metabolism to regulate its concentration in endometrial tissues is essential for the successful implantation and maintenance of pregnancy. In this study, we evaluated the expression of mRNA for 20alpha-hydroxysteroid dehydrogenase (20alpha-HSD), a key enzyme which converts progesterone to a biologically inactive metabolite, in human endometrial tissues and cultured endometrial stromal cells as well as decidua and chorionic tissues of early pregnancy. The level of 20alpha-HSD mRNA expression in secretory phase endometrium was significantly higher than that in proliferative phase endometrium and chorionic tissues. The expression level in decidual tissue was also significantly higher than that in chorionic tissue. In cultured endometrial stromal cells, 20alpha-HSD mRNA expression was slightly enhanced at a lower progesterone concentration of 0.01 micromol/l, and an increase in its expression was significantly suppressed at higher concentrations of 1 micromol/l or greater. No effect on the gene expression was seen in cultured endometrial stromal cells with various concentrations of 17beta-estradiol. These results suggest that progesterone itself contributes to the regulation of local progesterone concentration through 20alpha-HSD levels in endometrial stromal cells at peri implantation periods. PMID- 12733715 TI - Novel splicing events of untranslated first exons in human estrogen receptor alpha (ER alpha) gene. AB - In order to analyze the structures of the 5'-untranslated region of estrogen receptor alpha (ER alpha) mRNA in human uterine endometrium (Em), total RNA from Em was analyzed by 5'-rapid amplification of the cDNA ends method with antisense primer located on exon 1 of human ER alpha gene. Three isoforms of 5'-RACE clones were obtained: ER alpha mRNAs containing exon (A) (the upstream region of exon 1), exon C, and exons F-E2 (we adopted the nomenclature of 5'-untranslated exons of the Gannon group). The results imply that the major isoforms of ER alpha mRNA expressed in Em are these three isoforms. Moreover, reverse transcription polymerase chain reaction (RT-PCR) analysis was carried out on Em, ovary (Ov) and liver (Li) mRNAs to detect the novel isoforms of ER alpha mRNA in these tissues, using sense primers located on exons (A), B, C, F, and E1, and antisense primer located on exon 1. As a result, in addition to the previously reported ER alpha mRNA isoforms containing exons (A), B, C, F-E2 and E1-E2 on exon 1, we identified two novel isoform mRNAs in which exons F and E1 were directly spliced onto exon 1. Differential distributions of these isoforms of ER alpha mRNAs in Em, Ov and Li were demonstrated by RT-PCR-Southern blot analysis. These results, together with the previous reports by others, indicate that there are at least ten isoforms of ER alpha mRNA containing different 5'-untranslated regions, exons (A), B, C, D, T1-T2, T1, F-E2, F, E1-E2 and E1, expressed in human, and that these are involved in tissue specific expression of the gene. PMID- 12733717 TI - TSH-receptor antibody measurement in patients with various thyrotoxicosis and Hashimoto's thyroiditis: a comparison of two two-step assays, coated plate ELISA using porcine TSH-receptor and coated tube radioassay using human recombinant TSH receptor. AB - The aim of this study was to compare two two-step assays, a new coated plate (CP) ELISA assay (TRAb ELISA) using purified porcine TSH-receptors (pTSH-R) and a coated tube assay (CT) using recombinant human TSH-receptors (hTSH-R) (DYNO(R) test TRAK human). The same serum samples were used for the determination by both assays in patients with 100 untreated Graves' disease (GD), 30 silent thyroiditis (ST), 10 subacute thyroiditis (SAT) and 87 Hashimoto's thyroiditis (HT). In sera from patients with untreated GD, pTBII and hTBII were positive in nearly all cases except the same one, whereas the thirty sera from the ST had positive values of pTBII in one case and of hTBII in 4 cases. In the one ST case of both pTBII and hTBII positive, hyperthyroidism developed following ST, although the remaining ST cases including the three hTBII-positive cases were not followed by hyperthyroidism after ST attack. A positive value of hTBII was observed in one of 10 patients with SAT, whereas none of them was pTBII positive. In the 87 patients with HT, positive values of pTBII were recognized in 9 patients, whereas hTBII is positive in 10 patients. Serum TSAb and TSBAb activities were analyzed in the hTBII positive 7 patients. As a result, TSAb was all positive except one and TSBAb positive in 4 cases. Since there is no significant difference in the sensitivity and specificity between the two assays in the differentiation of thyrotoxicosis as well as the frequency of finding positive values in patients with HT, it is reasonable to conclude that the clear advantage of sensitivity for clinical application in the new CP and CT assays may be derived from the coated plate or coated tube assay itself, which probably excludes the effect of anti-TSH antibodies and HAMA, and is unrelated to the use of human or porcine TSH receptors. PMID- 12733718 TI - Two key proteins of the vitamin D endocrine system come into crystal clear focus: comparison of the X-ray structures of the nuclear receptor for 1alpha,25(OH)2 vitamin D3, the plasma vitamin D binding protein, and their ligands. PMID- 12733719 TI - Mechanical loading stimulates dentin matrix protein 1 (DMP1) expression in osteocytes in vivo. AB - Dentin matrix protein 1 (DMP1) was originally postulated to be dentin specific. Further analysis showed that DMP1 is also expressed in mature cartilage and bone. In bone tissue, DMP1 is expressed predominantly in late osteoblasts and osteocytes. DMP1 belongs to the SIBLING (Small Integrin Binding Ligand N-linked Glycoprotein) family of cellular matrix proteins that also includes osteopontin, bone sialoprotein, dentin sialophosphoprotein, and others. In this study, we examined the effect of mechanical loading on expression of DMP1 mRNA and DMP1 protein in alveolar bone in the mouse tooth movement model by in situ hybridization and immunocytochemistry. The expression of DMP1 mRNA was determined quantitatively in mechanically loaded and control sites of dento-alveolar tissue at several time points from 6 h to 7 days after loading. The tooth movement model allows simultaneous evaluation of bone resorption and bone formation sites. Expression of DMP1 mRNA in osteocytes increased 2-fold as early as 6 h after treatment in both the bone formation and bone resorption sites. After 4 days, DMP1 expression in osteocytes increased to a maximum of 3.7-fold in the bone formation sites and 3.5-fold in the resorption sites. Osteoblasts responded in the opposite manner and showed a transient 45% decrease of DMP1 mRNA in bone formation sites and a constant decrease of DMP1 mRNA during the entire course of treatment in the bone resorption sites, with a peak inhibition of 67% at day 2. By immunocytochemistry using a C-terminal region peptide antibody to DMP1, we found that there was a transient decrease in immunoreactivity at 3 days after treatment on both the formation side and the resorption side compared with the matched contralateral control tissue. However by 7 days of loading, there was a dramatic increase in DMP1 protein immunoreactivity on both the formation side and the resorption side. These results represent changes in epitope availability using this antibody or true changes in protein levels. The observations imply that the DMP1 protein is undergoing dynamic changes in either synthesis or other protein/matrix interaction after mechanical loading of alveolar bone. The findings indicate that DMP1 is involved in the responses of osteocytes and osteoblasts to mechanical loading of bone. These results support the hypothesis that osteocytes alter their matrix microenvironment in response to mechanical loading. PMID- 12733720 TI - p21(WAF1/CIP1) acts as a brake in osteoblast differentiation. AB - Continuous fibroblast growth factor signaling inhibits the differentiation of primary osteoblasts and osteoblastic cell lines. We studied the expression of several cell cycle regulatory molecules in response to fibroblast growth factor, and found that fibroblast growth factor strongly upregulates the expression of p21(WAF1/CIP1), a CDK inhibitor that has also been implicated in the regulation of apoptosis and cell differentiation. To test the hypothesis that p21 mediated the fibroblast growth factor effects on osteoblasts, we studied the differentiation of primary osteoblasts and osteoblastic cell lines derived from p21 null mice in the presence or absence of fibroblast growth factor. While the results obtained indicate that p21 is not the major mediator of the inhibition of osteoblast differentiation by fibroblast growth factor, we found that p21 per se acts as a brake on osteoblast proliferation and differentiation. p21 is strongly downregulated during differentiation and is highly expressed in osteoblastic cell lines expressing activated FGFR2, which do not differentiate. p21 null osteoblasts differentiate faster than wild-type cells, are more susceptible to the differentiation-promoting action of BMP-2, and undergo increased differentiation-related apoptosis. Furthermore, transient overexpression of p21 from an adenovirus vector delayed the onset of differentiation both in wild-type and in p21 null osteoblasts. These results highlight a new function for p21 in osteoblast differentiation. PMID- 12733721 TI - Receptor tyrosine kinases inhibit bone morphogenetic protein-Smad responsive promoter activity and differentiation of murine MC3T3-E1 osteoblast-like cells. AB - Growth factors such as fibroblast growth factor-2 (FGF-2) and epidermal growth factor (EGF) that activate extracellular signal-regulated kinases (ERKs) through receptor tyrosine kinases (RTKs) stimulate proliferation but suppress differentiation of osteoblasts. To study the mechanism of this inhibitory action of these growth factors on osteoblastic differentiation, we evaluated Smad1 transactivity in MC3T3-E1 osteoblast-like cells by reporters of promoter activity of mouse Smad6, an early response gene to bone morphogenetic proteins (BMPs). FGF 2 and EGF inhibited alkaline phosphatase activity and Smad6 promoter activity stimulated by BMP-2. Overexpression of constitutively active MEK by adenovirus mimicked, but that of dominant negative Ras or treatment with a MEK1 inhibitor, PD098059, reversed, the inhibitory effects of these growth factors on both activities. These effects are mediated by BMP-responsive elements (BMPREs) on Smad6 promoter, because an artificial reporter driven by three tandem BMPREs gave similar results, and these effects were all abolished when the BMPREs were mutated. RTK-ERK activation inhibited the promoter activity even when BMP signal was mediated by a mutant Smad1, which lacks phosphorylation sites by ERKs, or by a Smad1 fused to Gal4 DNA binding domain, which constitutively localizes in the nucleus. These results show that the RTK-Ras-ERK pathway suppresses BMP signal by interfering with Smad1 transactivity. Because direct phosphorylation of Smad1 by ERKs is not required for the inhibition, other transcriptional factors that are phosphorylated by ERKs might be involved in the regulation of osteoblastic differentiation by ERKs. PMID- 12733722 TI - Paracrine overexpression of IGFBP-4 in osteoblasts of transgenic mice decreases bone turnover and causes global growth retardation. AB - Insulin-like growth factor binding protein 4 (IGFBP-4) is abundantly expressed in bone and is generally believed to function as an inhibitor of IGF action. To investigate the function of locally produced IGFBP-4 in bone in vivo, we targeted expression of IGFBP-4 to osteoblasts using a human osteocalcin promoter to direct transgene expression. IGFBP-4 protein levels in calvaria of transgenic (OC-BP4) mice as measured by Western ligand blot were increased 25-fold over the endogenous level. Interestingly, levels of IGFBP-5 were decreased in the OC-BP4 mice, possibly because of a compensatory alteration in IGF-1 action. Morphometric measurements showed a decrease in femoral length and total bone volume in transgenic animals compared with the controls. Quantitative histomorphometry at the distal femur disclosed a striking reduction in bone turnover in the OC-BP4 mice. Osteoblast number/bone length and bone formation rate/bone surface in OC BP4 mice were approximately one-half that seen in control mice. At birth, OC-BP4 mice were of normal size and weight but exhibited striking postnatal growth retardation. Organ allometry (mg/g body weight) analysis revealed that, whereas most organs exhibited a proportional reduction in weight, calvarial and femoral wet weights were disproportionally small (approximately 70% and 80% of control, respectively). In conclusion, paracrine overexpression of IGFBP-4 in the bone microenvironment markedly reduced cancellous bone formation and turnover and severely impaired overall postnatal skeletal and somatic growth. We attribute these effects to the sequestration of IGF-1 by IGFBP-4 and consequent impairment of IGF-1 action in skeletal tissue. PMID- 12733723 TI - Distinct phases of coordinated early and late gene expression in growth plate chondrocytes in relationship to cell proliferation, matrix assembly, remodeling, and cell differentiation. AB - Although much has been learned about growth plate development and chondrocyte gene expression during cellular maturation and matrix remodeling in the mouse, there has been a limited study of the interrelationships of gene expression between proteinases, growth factors, and other regulatory molecules in the mouse and in other species. Here we use RT-PCR of sequential transverse sections to examine the expression profiles of genes involved in chondrocyte growth, differentiation, matrix assembly, remodeling, and mineralization in the bovine proximal tibial growth plate. Specifically, we studied the expression of genes encoding COL2A1 and COL10A1, the latter a marker of cellular hypertrophy, the matrix metalloproteinases (MMP), MMP-13 and MMP-9, as well as the transcriptional factors, Sox9 and Cbfa1, the growth factors basic fibroblast growth factor (bFGF), parathyroid hormone-related peptide (PTHrP), transforming growth factor (TGF)beta1, and beta2, Indian hedgehog (Ihh), and the matrix protein osteocalcin. These were analyzed in relationship to cell division defined by cyclin B2 expression. Two peaks of gene expression activity were observed. One was transient, limited, and located immediately before and at the onset of cyclin B2 expression in the early proliferative zone. The other was generally much more pronounced and was located in the early hypertrophic zone. The upregulation of expression of COL2A1, its transcriptional activator Sox9, osteocalcin, MMP-13, and TGFbeta2 was observed immediately before and at the onset of cyclin B2 expression and also in the hypertrophic zones. The upregulation of COL10A1, Cbfa1, MMP-9, TGFbeta-1, and Ihh gene expression was associated exclusively with the terminal differentiation of chondrocytes at the time of mineral formation in the extracellular matrix. In contrast, bFGF and PTHrP expression was observed in association with the onset of cyclin B2 expression and hypertrophy. This initial cluster of gene expression associated predominantly with matrix assembly and onset of cell proliferation is therefore characterized by expression of regulatory molecules distinct from those involved at hypertrophy. Together these results identify separate phases of coordinated gene expression associated with the development of the physis in endochondral bone formation. PMID- 12733724 TI - Sustained antiresorptive effects after a single treatment with human recombinant osteoprotegerin (OPG): a pharmacodynamic and pharmacokinetic analysis in rats. AB - Osteoprotegerin (OPG) is a naturally occurring negative regulator of osteoclast differentiation, activation, and survival. We created a recombinant form of human OPG (rhOPG), with a sustained serum half-life, to achieve prolonged antiresorptive activity. This study describes the rapid and sustained antiresorptive effects that are achieved with a single treatment with rhOPG. Male Sprague-Dawley rats (10 weeks old) were given a single bolus intravenous injection of vehicle (PBS) or rhOPG (5 mg/kg). PBS- and rhOPG-treated rats (n = 6/group) were killed at 0, 0.5, 1, 2, 5, 10, 20, and 30 days post-treatment. rhOPG-treated rats were compared with their age-matched controls. The main pharmacologic effect of rhOPG was a rapid (24 h) reduction in osteoclast surface in the tibia, which reached a nadir on days 5 and 10 (95% reduction vs. vehicle controls). Osteoclast surface remained significantly reduced 30 days after the single treatment with rhOPG. Tibial cancellous bone volume was significantly increased within 5 days of rhOPG treatment (23%) and reached a peak increase of 58% on day 30. Femoral bone mineral density was significantly increased in rhOPG treated rats on days 10 and 20. Pharmacokinetic analysis revealed that serum concentrations of rhOPG remained at measurable levels throughout the 30-day study. These data show that a single intravenous injection of rhOPG in young growing rats causes significant gains in bone volume and density, which are associated with rapid and sustained suppression of osteoclastic bone resorption. PMID- 12733726 TI - Relationships between pharmacokinetics and rate of bone turnover after intravenous bisphosphonate (olpadronate) in patients with Paget's disease of bone. AB - Bisphosphonates are the treatment of choice of Paget's disease, but variable responses have been reported, and despite the availability of potent bisphosphonates, biochemical remission is not achieved in a substantial number of patients. This may, in part at least, be because of the influence of pharmacokinetics of bisphosphonates on their pharmacodynamics. That is the response of bone turnover to treatment. To address this issue, we examined the pharmacokinetics and pharmacodynamics of the bisphosphonate olpadronate given intravenously to 75 patients with Paget's disease, using a specific assay for olpadronate concentrations in serum and urine. The skeletal uptake of olpadronate varied greatly among patients and ranged between 10% and 90% of the administered dose. The two major determinants of skeletal uptake were renal function and prevalent rate of bone turnover. Serum and urinary data were well described by a physiology-based four-compartment pharmacokinetic model that takes into account the distribution of the bisphosphonate in the bone and its subsequent elimination. Bone turnover was suppressed to well within the normal range in virtually all patients. This, together with the absence of resolution of effect during 1 year, does not allow the construction of an adequate integrated pharmacokinetic/pharmacodynamic model. However, the pharmacokinetic model, described for the first time in Paget's disease, can accurately simulate the amount of bisphosphonate delivered to the skeleton with different dose regimens as well as the amount still present in bone after 1 year. Such approaches can lead to improved patient care and individualization of treatment of Paget's disease with bisphosphonates. PMID- 12733725 TI - The type I collagen fragments ICTP and CTX reveal distinct enzymatic pathways of bone collagen degradation. AB - Bone resorption may generate collagen fragments such as ICTP and CTX, which can be quantified in serum and/or urine by using specific immunoassays, and which are used as clinical markers. However, the relative abundance of ICTP and CTX varies according to the type of bone pathology, suggesting that these two fragments are generated through distinct collagenolytic pathways. In this study, we analyzed the release of ICTP and CTX from bone collagen by the proteinases reported to play a role in the solubilization of bone matrix. Cathepsin K released large amounts of CTX, but did not allow a detectable release of ICTP. Conversely, the matrix metalloproteinases (MMPs) MMP-2, -9, -13, or -14 released ICTP, but did not allow a detectable release of CTX. Next we analyzed the release of ICTP and CTX from bone explants cultured in the presence of well-established inhibitors of these proteinases and of matrix solubilization. An inhibitor of cysteine proteinases including cathepsin K, inhibited the release of CTX, but not the release of ICTP. MMP inhibitors inhibited the release of ICTP, but also that of CTX, in agreement with the putative role of MMPs in the initiation of bone resorption in addition to matrix solubilization. Similarly the treatment of mice bearing bone metastasis with an MMP inhibitor led to a significant reduction of serum ICTP and CTX, and osteolytic lesions. We conclude that the generation of ICTP and CTX depends on different collagenolytic pathways. This finding may explain why these two markers may discriminate between different bone pathologies. PMID- 12733727 TI - Effect of 8-month vertical whole body vibration on bone, muscle performance, and body balance: a randomized controlled study. AB - Recent animal studies have given evidence that vibration loading may be an efficient and safe way to improve mass and mechanical competence of bone, thus providing great potential for preventing and treating osteoporosis. Randomized controlled trials on the safety and efficacy of the vibration on human skeleton are, however, lacking. This randomized controlled intervention trial was designed to assess the effects of an 8-month whole body vibration intervention on bone, muscular performance, and body balance in young and healthy adults. Fifty-six volunteers (21 men and 35 women; age, 19-38 years) were randomly assigned to the vibration group or control group. The vibration intervention consisted of an 8 month whole body vibration (4 min/day, 3-5 times per week). During the 4-minute vibration program, the platform oscillated in an ascending order from 25 to 45 Hz, corresponding to estimated maximum vertical accelerations from 2 g to 8 g. Mass, structure, and estimated strength of bone at the distal tibia and tibial shaft were assessed by peripheral quantitative computed tomography (pQCT) at baseline and at 8 months. Bone mineral content was measured at the lumbar spine, femoral neck, trochanter, calcaneus, and distal radius using DXA at baseline and after the 8-month intervention. Serum markers of bone turnover were determined at baseline and 3, 6, and 8 months. Five performance tests (vertical jump, isometric extension strength of the lower extremities, grip strength, shuttle run, and postural sway) were performed at baseline and after the 8-month intervention. The 8-month vibration intervention succeeded well and was safe to perform but had no effect on mass, structure, or estimated strength of bone at any skeletal site. Serum markers of bone turnover did not change during the vibration intervention. However, at 8 months, a 7.8% net benefit in the vertical jump height was observed in the vibration group (95% CI, 2.8-13.1%; p = 0.003). On the other performance and balance tests, the vibration intervention had no effect. In conclusion, the studied whole body vibration program had no effect on bones of young, healthy adults, but instead, increased vertical jump height. Future human studies are needed before clinical recommendations for vibration exercise. PMID- 12733728 TI - Randomized trial of physical activity and calcium supplementation on bone mineral content in 3- to 5-year-old children. AB - A meta-analysis of adult exercise studies and an infant activity trial show a possible interaction between physical activity and calcium intake on bone. This randomized trial of activity and calcium supplementation was conducted in 239 children aged 3-5 years (178 completed). Children were randomized to participate in either gross motor or fine motor activities for 30 minutes/day, 5 days per week for 12 months. Within each group, children received either calcium (1000 mg/day) or placebo. Total body and regional bone mineral content by DXA and 20% distal tibia measurements by peripheral quantitative computed tomography (pQCT) were obtained at 0 and 12 months. Three-day diet records and 48-h accelerometer readings were obtained at 0, 6, and 12 months. Higher activity levels were observed in gross motor versus fine motor activity groups, and calcium intake was greater in calcium versus placebo (1354 +/- 301 vs. 940 +/- 258 mg/day, p < 0.001). Main effects of activity and calcium group were not significant for total body bone mineral content or leg bone mineral content by DXA. However, the difference in leg bone mineral content gain between gross motor and fine motor was more pronounced in children receiving calcium versus placebo (interaction, p = 0.05). Children in the gross motor group had greater tibia periosteal and endosteal circumferences by pQCT compared with children in the fine motor group at study completion (p < 0.05). There was a significant interaction (both p < or = 0.02) between supplement and activity groups in both cortical thickness and cortical area: among children receiving placebo, thickness and area were smaller with gross motor activity compared with fine motor activity, but among children receiving calcium, thickness and area were larger with gross motor activity. These findings indicate that calcium intake modifies the bone response to activity in young children. PMID- 12733729 TI - Nulliparity and fracture risk in older women: the study of osteoporotic fractures. AB - Whether nulliparity increases fracture risk is unclear from prior studies, which are limited by small samples or lack of measured bone mineral density. No study has evaluated whether the effect of parity differs by skeletal site. We prospectively analyzed the relationship of parity to the risk of incident nontraumatic hip, spine, and wrist fractures in 9704 women aged 65 years or older participating in the Study of Osteoporotic Fractures to determine if parity reduces postmenopausal fracture risk, and if so, if this risk reduction is (1) greater at weight-bearing skeletal sites and (2) independent of bone mineral density. Parity was ascertained by self-report. Incident hip and wrist fractures were determined by physician adjudication of radiology reports (mean follow-up, 9.8 years) and spine fractures by morphometric criteria on serial radiographs. The relationship of parity to hip and wrist fracture was assessed by proportional hazards models. Spine fracture risk was evaluated by logistic regression. Compared with parous women, nulliparous women (n = 1835, 19%) had an increased risk of hip and spine, but not wrist, fractures. In multivariate models, parity remained a significant predictor only for hip fracture. Nulliparous women had a 44% increased risk of hip fractures independent of hip bone mineral density (hazards ratio, 1.44; 95% CI, 1.17-1.78). Among parous women, each additional birth reduced hip fracture risk by 9% (p = 0.03). Additionally, there were no differences in mean total hip, spine, or radial bone mineral density values between nulliparous and parous women after multivariate adjustment. In conclusion, childbearing reduces hip fracture risk by means that may be independent of hip bone mineral density. PMID- 12733730 TI - Fracture risk after bilateral oophorectomy in elderly women. AB - Elderly women with the lowest serum estrogen levels are at the greatest risk of bone loss and fractures, but it is controversial whether the ovaries contribute to estrogen production after menopause, and therefore, whether bilateral oophorectomy in postmenopausal women might have adverse skeletal effects. To address this potential problem, we estimated long-term fracture risk among 340 postmenopausal Olmsted County, MN, women who underwent bilateral oophorectomy for a benign ovarian condition in 1950-1987. In over 5632 person-years of follow-up (median, 16 years per subject), 194 women experienced 516 fractures (72% from moderate trauma). Compared with expected rates, there was a significant increase in the risk of any osteoporotic fracture (moderate trauma fractures of the hip, spine, or distal forearm; standardized incidence ratio [SIR], 1.54; 95% CI, 1.29 1.82) but almost as large an increase in fractures at other sites (SIR, 1.35; 95% CI, 1.13-1.59). In multivariate analyses, the independent predictors of overall fracture risk were age, anticonvulsant or anticoagulant use for > or = 6 months, and a history of alcoholism or prior osteoporotic fracture; obesity was protective. Estrogen replacement therapy was associated with a 10% reduction in overall fracture risk (hazard ratio [HR], 0.90; 95% CI, 0.64-1.28) and a 20% reduction in osteoporotic fractures (HR, 0.80; 95% CI, 0.52-1.23), but neither was statistically significant. The increase in fracture risk among women who underwent bilateral oophorectomy after natural menopause is consistent with the hypothesis that androgens produced by the postmenopausal ovary are important for endogenous estrogen production that protects against fractures. PMID- 12733731 TI - Can novel clinical densitometric techniques replace or improve DXA in predicting bone strength in osteoporosis at the hip and other skeletal sites? AB - New peripheral techniques are now available for the diagnosis of osteoporosis, but their value in the clinical management of the disease remains controversial. This study tests the hypothesis that peripheral quantitative computed tomography (pQCT) at the distal radius and/or quantitative ultrasound (QUS) at the calcaneus can serve as replacement or improvement of current methodology (QCT and DXA) for predicting bone strength at the hip and other sites. In 126 human cadavers (age, 80.2 +/- 10.4 years), DXA of the femur, spine, and radius and pQCT of the radius were acquired with intact soft tissues. QCT (spine) and QUS (calcaneus) were performed ex situ in degassed specimens. Femoral failure loads were assessed in side impact and vertical loading. Failure loads of the thoracolumbar spine were determined at three levels in compression and those of the radius by simulating a fall. Site-specific DXA explained approximately 55% of the variability in femoral strength, whereas pQCT and QUS displayed a lower association (15-40%). QUS did not provide additional information on mechanical strength of the femur, spine, or radius. All techniques displayed similar capability in predicting a combined index of failure strength at these three sites, with only QUS exhibiting significantly lower associations than other methods. These experimental results suggest that clinical assessment of femoral fracture risk should preferably rely on femoral DXA, whereas DXA, QCT, and pQCT display similar capability of predicting a combined index of mechanical strength at the hip, spine, and radius. PMID- 12733732 TI - Children and the risk of fractures caused by oral corticosteroids. AB - Oral corticosteroids are known to increase the risk of fracture in adults, but their effects in children remain uncertain. The medical records of general practitioners in the United Kingdom (from the General Practice Research Database) were used to estimate the incidence rates of fracture of children ages 4-17 years taking oral corticosteroids (n = 37,562) and of control children taking nonsystemic corticosteroids (n = 345,748). Each child with a fracture (n = 22,846) was subsequently matched by age, sex, practice, and calendar time to one child without a fracture. The average duration of treatment was 6.4 days (median, 5 days). The risk of fracture was increased in children with a history of frequent use of oral corticosteroids; children who received four or more courses of oral corticosteroids had an adjusted odds ratio (OR) for fracture of 1.32 (95% CI, 1.03-1.69). Of the various fracture types, the risk of humerus fracture was doubled in children who received four or more courses of oral corticosteroids (adjusted OR, 2.17 [1.01-4.67]). Fracture risk was also increased among children using 30 mg prednisolone or more each day (adjusted OR for fracture, 1.24 [1.00 1.52]) and among those receiving four or more courses of oral corticosteroids (OR, 1.32 [1.03-1.69]). Children who stopped taking oral corticosteroids had a comparable risk of fracture to those in the control group. Our findings suggest that children who require more than four courses of oral corticosteroid as treatment for underlying disease are at increased risk of fracture. It is not entirely clear whether this relates directly to oral corticosteroid use or the underlying disease and its severity. Irrespective of these issues, this group of children is at increased risk of fracture. PMID- 12733733 TI - Prevention and treatment of glucocorticoid-induced osteoporosis: a comparison of calcitriol, vitamin D plus calcium, and alendronate plus calcium. AB - High-dose corticosteroids, used for many medical conditions, are associated with rapid bone loss from sites such as the vertebrae, and compression fractures can be observed within months. Recent trials suggest treatment with bisphosphonates or active vitamin D analogs can reduce bone loss and the risk of fracture associated with glucocorticoids, but few studies have directly compared such agents. We conducted a randomized, multicenter, open-label trial to compare the efficacy of alendronate, calcitriol, and simple vitamin D in prevention and treatment of glucocorticoid-induced bone loss. A total of 195 subjects (134 females and 61 males) commencing or already taking glucocorticoids were randomized to one of three groups: calcitriol, 0.5 to 0.75 microg/day; simple vitamin D (ergocalciferol, 30,000 IU weekly) plus calcium carbonate (600 mg daily); or alendronate, 10 mg/day plus calcium carbonate (600 mg daily). Over 2 years, mean lumbar bone mineral density change was +5.9% with alendronate, -0.5% with ergocalciferol, and -0.7% with calcitriol (p < 0.001). At the femoral neck, there was no significant difference in bone mineral density change between the treatments over 2 years: alendronate (+0.9%), ergocalciferol (-3.2%), and calcitriol (-2.2%). Lumbar bone loss varied according to whether patients were starting or receiving chronic glucocorticoids, and there was a significant treatment x prior glucocorticoid use interaction effect. Six of 66 calcitriol subjects, 1 of 61 ergocalciferol subjects, and 0 of 64 alendronate subjects sustained new vertebral fractures. These data do not suggest any difference between simple vitamin D and calcitriol but do show that alendronate was superior to either treatment for glucocorticoid induced bone loss. PMID- 12733734 TI - A longitudinal study of the effect of subcutaneous estrogen replacement on bone in young women with Turner's syndrome. AB - It is desirable that young women with primary ovarian failure achieve normal peak bone mass to reduce the subsequent risk of osteoporosis, and that there are management strategies to replace bone that is already lost. While estrogen (E2) is generally considered to prevent bone loss by suppressing bone resorption, it is now recognized that estrogen also exerts an anabolic effect on the human skeleton. In this study, we tested whether estrogen could increase bone mass in women with primary ovarian failure. We studied the mechanism underlying this by analyzing biochemical markers of bone turnover and iliac crest biopsy specimens obtained before and 3 years after E2 replacement. Twenty-one women with Turner's syndrome, aged 20-40 years, were studied. The T scores of bone mineral density at lumbar spine and proximal femur at baseline were -1.4 and -1.1, respectively. Hormone replacement was given as subcutaneous E2 implants (50 mg every 6 months) with oral medroxy progesterone. Serum E2 levels increased incrementally from 87.5 pM at baseline to 323, 506, 647, and 713 pM after 6 months and 1, 2, and 3 years of hormone replacement therapy (HRT), respectively. The bone mineral density at the lumbar spine and proximal femur increased after 3 years to T scores of -0.2 and -0.4, respectively. The cancellous bone volume increased significantly from 13.4% to 18.8%. There was a decrease in activation frequency, but the active formation period was increased by HRT. There was a significant increase in the wall thickness from 33.4 microm at baseline to 40.9 microm after 3 years of HRT, reflecting an increase in bone formed at individual remodeling units. Although there was an early increase in biochemical markers of bone formation, these declined thereafter. Our results show that estrogen is capable of exerting an anabolic effect in the skeleton of young women with Turner's syndrome and low bone mass. PMID- 12733736 TI - Radioguided parathyroidectomy for recurrent hyperparathyroidism caused by forearm graft hyperplasia. AB - One of the surgical options for symptomatic secondary hyperparathyroidism is a total parathyroidectomy with forearm implantation. Recurrence can occur and is most likely caused by hyperplasia of the small fragments of parathyroid tissue implanted in the forearm muscle. Forearm graft hyperplasia can be detected using Tc-99m sestamibi scanning of the forearm, which can show abnormal enhancement at the former graft site. In this report, we present the case of a 49-year-old gentleman with recurrent hyperparathyroidism caused by hyperplasia of forearm graft fragments. Unfortunately, no sutures or clips were placed at his initial surgery to identify the location of the parathyroid tissue in the forearm. Thus, we describe the first reported use of radioguided techniques using Tc-99m sestamibi injection and intraoperative gamma probe to localize parathyroid fragments in the forearm muscle. During our initial exploration, we found that injection of the tracer in the operative arm leads to prohibitively high levels of background activity. During a second exploration, the tracer was injected in the lower extremity, minimizing the background in the forearm and allowing the gamma probe to clearly identify two areas of abnormal parathyroid tissue. The intraoperative radioprobe allowed quick identification and removal of the abnormal parathyroid tissue in a case that was made particularly challenging by the absence of marking sutures. PMID- 12733735 TI - Visual identification of vertebral fractures in osteoporosis using morphometric X ray absorptiometry. AB - Visual identification of vertebral fractures from spinal radiographs (visual XR) makes use of the reader's expertise in ruling out non-fracture deformities or normal variants. Scan images of the spine acquired by DXA may be analyzed quantitatively (morphometric X-ray absorptiometry [quantitative MXA]) or visually (visual MXA). The aims of this study were to compare visual and quantitative MXA with visual XR for the identification of vertebral fractures. Spinal radiographs and MXA scans were acquired at baseline and 1 year in 70 women referred with osteoporosis. These were assessed visually by two expert readers (observer A, a radiologist; observer B, a physician with expertise in osteoporosis) for evidence of prevalent and incident vertebral fractures. Observer C (a radiographer with expertise in vertebral morphometry) performed visual and quantitative assessments of the MXA scans. Visual assessment of spinal radiographs by observer A was used as the gold standard for comparison of methods. Sensitivity for the identification of prevalent fractures by MXA was best for visual MXA by observer A (92%), whereas quantitative MXA had the lowest sensitivity (82%). Specificity was >90% for both visual and quantitative MXA. Kappa scores for agreement for identification of prevalent fractures between visual XR (observer A) and visual MXA (all three observers), and between visual XR and visual MXA performed by reader B were similar (kappa = 0.85-0.87). Agreement with visual XR performed by observer A was slightly lower for quantitative MXA (kappa = 0.77). Interobserver agreement between the two expert readers (observers A and B) was the same for both visual XR and visual MXA (kappa = 0.86). Seven incident vertebral fractures were identified in four patients at follow-up. All four patients were identified by visual MXA, and three patients were identified by quantitative MXA. Observers A and B identified all seven incident fractures by visual MXA, and observer C missed one fracture that was also missed by quantitative MXA. An incident fracture of vertebra T6 was excluded from analysis by quantitative MXA because of poor image quality. We conclude that visual identification of vertebral fractures from MXA scans is superior to quantitative assessment. Used as a screening tool for conventional radiography, this approach could help reduce the radiation dose to the patient in the diagnosis and monitoring of osteoporosis. PMID- 12733737 TI - IGF-I-deficient mice: role in skeletal adaptation to load. PMID- 12733738 TI - Dietary fat, obesity, and health--from theory to practice. Proceedings of the Mark Bieber Memorial Symposium, presented at the 93rd AOCS annual meeting in Montreal, Canada, May 2002. PMID- 12733739 TI - Genetics of obesity: more complicated than initially thought. AB - During the past several decades, there has been an explosion in the prevalence of obesity. Since our genes have not changed appreciably during that time, it stands to reason that the present epidemic is caused by our pervasive obesigenic environment, in which excess caloric intake and decreased physical activity conspire with one another. Despite an obesigenic environment, humans have great variability in their susceptibility to obesity, which is determined in large part by genetics. Current evidence suggests that genetic susceptibility to human obesity is the result of multiple genes, each with a modest effect, that inter act with each other and with environmental provocations. Elucidation of obesity susceptibility genes through genome-wide and candidate gene approaches provides great promise in ultimately determining the genetic underpinnings of obesity. Further research will translate these new insights on the pathophysiological basis of obesity into new medications and diagnostic tests. PMID- 12733740 TI - Sedentary lifestyle and risk of obesity and type 2 diabetes. AB - Obesity and type 2 diabetes have reached epidemic proportions in the United States. It is well-established that increasing physical activity plays an important role in reducing risk of obesity and diabetes. Few studies, however, have examined the association between sedentary behaviors such as prolonged television (TV) watching and obesity and diabetes. Using data from a large prospective cohort study, the Health Professionals' Follow-up Study, we have demonstrated that increasing TV watching is strongly associated with obesity and weight gain, independent of diet and exercise. Also, prolonged TV watching is associated with a significantly increased risk of type 2 diabetes. Men who watched TV more than 40 h per week had a nearly threefold increase in the risk of type 2 diabetes compared with those who spent less than 1 h per week watching TV. The increased risk was not entirely explained by the decreased physical activity and unhealthy eating patterns associated with TV watching. Thus, public health campaigns to reduce the risk of obesity and type 2 diabetes should promote not only increasing exercise levels but also decreasing sedentary behaviors, especially prolonged TV watching. PMID- 12733741 TI - The role of energy density. AB - Dietary energy density (ED) appears to have a major influence on the regulation of food intake and body weight. If people consume a fixed weight of food each day, then high-ED diets should be associated with high energy intakes and with overweight. In contrast, low-ED diets should result in lower daily energy intakes and therefore weight loss. For this approach to work, low-ED foods must be as palatable as high-ED foods and, calorie for calorie, have a greater satiating power. Each of those assumptions is debatable. Dietary ED depends chiefly on the water content of foods. As a rule, high-ED foods are more palatable but less satiating, whereas low-ED foods are more satiating but less palatable. Consumer preferences for high-ED foods can be explained in terms of good taste, low cost, and convenience. Low-ED foods, such as fresh produce, provide less energy per unit cost than do high-ED foods, which often contain added sugars and fats. Poverty and obesity may well be linked through the habitual consumption of a low cost, high-ED diet. PMID- 12733743 TI - Dietary fat and body weight control. AB - The global obesity epidemic has heightened the debate about dietary factors contributing to weight gain. Media stories have promulgated the notion that obesity has increased despite reductions in dietary fat intake. Some have even speculated that lower dietary fat levels may be driving the rapid rise in weight gain within the population. A close examination of the science reveals a different picture and supports the hypothesis that dietary fat, within the context of the total dietary composition consumed by many populations, promotes obesity. Hence, dietary fat control is still an important strategy as part of an overall approach to body weight management in our modern environment. Dietary fat increases the energy density of foods. Abundant evidence from preclinical and clinical studies indicates that fat promotes excess energy intake and positive energy balance. Dietary fat does not promote its own oxidation in the body and is stored efficiently, promoting a positive fat balance. Thus, both the behavioral and metabolic responses to dietary fat increase the probability of positive energy balance and body fat gain. Restoring fat balance when consuming diets rich in fat requires increasing the size of the body fat mass, increasing physical activity, or reducing dietary fat intake. Numerous epidemiologic, preclinical, and controlled clinical studies have shown that body fat is positively associated with dietary fat intake and that dietary fat manipulation leads to appropriate changes in body fat mass. Finally, data from the National Weight Control Registry, a database of > 3000 individuals who have successfully maintained a substantial weight loss, indicate that moderating dietary fat intake is a key strategy for long-term management of body weight. PMID- 12733742 TI - Dietary glycemic index and the regulation of body weight. AB - Prevalence rates of overweight and obesity have risen precipitously in the United States and other developed countries since the 1960s, despite comprehensive public health efforts to combat this problem. Although considerable attention has been focused on decreasing dietary fat and increasing physical activity level, the potential relevance of the dietary glycemic index to obesity treatment has received comparatively little scientific notice. This review examines how the glycemic and insulinemic responses to diet may affect body weight regulation, and argues for the potential utility of low glycemic index diets in the prevention and treatment of obesity and related complications. PMID- 12733744 TI - Nutritional characteristics of DAG oil. AB - Excess calorie intake in industrialized countries has prompted development of fat substitutes and other lower-calorie dietary items to enhance health. DAG cooking oils, with a 1,3 configuration, taste and have the texture of commonly used TAG cooking oils. Because they are not hydrolyzed to 2-MAG in the gut, the absorption and metabolism of DAG oil differs from that of TAG. Among the physiological differences are lower postprandial lipemia and an increased proportion of FA being oxidized instead of stored. Preliminary studies suggest that these differences in energy partitioning between DAG and TAG may be usefully exploited to reduce the amount of fat stored from cooking oil and oil components of food items. Over 70 million bottles of DAG oil have been sold in Japan since its introduction in February 1999, and the product is being test-marketed in the United States. It is hoped that wider use of DAG oil may provide one additional means of preventing obesity. PMID- 12733745 TI - CLA and body weight regulation in humans. AB - CLA comprises a group of unsaturated FA isomers with a variety of biological effects in experimental animals. CLA reduces body fat accumulation in animal models and has been suggested to have significant effects on lipid and glucose metabolism, e.g., antidiabetic effects in obese Zucker rats. It has been proposed that the trans10-cis12 isomer is the active isomer associated with the antiobesity and insulin-sensitizing properties of CLA. The metabolic effects in humans in general, and isomer-specific effects specifically, are not well characterized. In a series of controlled studies in humans, we investigated the effects of CLA (given as the commercially available mixture of isomers and as the purified trans10-cis12 CLA isomer) on anthropometry, lipid and glucose metabolism, and markers of lipid peroxidation. Preliminary results indicate that CLA may slightly decrease body fat in humans also, particularly abdominal fat, but there is no effect on body weight or body mass index. There is no simultaneous improvement in lipid or glucose metabolism. Rather, the trans10 cis12 CLA isomer unexpectedly caused significant impairment of the peripheral insulin sensitivity as well as of blood glucose and serum lipid levels. In addition, CLA markedly elevated lipid peroxidation. Thus, the metabolic effects of CLA in humans seem complex; further studies, especially of isomer-specific effects and for longer time periods, are warranted. PMID- 12733746 TI - Role of dietary calcium and dairy products in modulating adiposity. AB - Dietary calcium plays a pivotal role in the regulation of energy metabolism. High calcium diets attenuate adipocyte lipid accretion and weight gain during overconsumption of an energy-dense diet and increase lipolysis and preserve thermogenesis during caloric restriction, thereby markedly accelerating weight loss. Our studies of the agouti gene demonstrate a key role for intracellular Ca2+ in regulating adipocyte lipid metabolism and TG storage. Increased intracellular Ca2+ resulting in stimulation of lipogenic gene expression, and lipogenesis and suppression of lipolysis resulting in adipocyte lipid filling and increased adiposity. Moreover, we recently demonstrated that the increased calcitriol produced in response to low-calcium diets stimulates adipocyte Ca2+ influx and, consequently, promotes adiposity. Accordingly, suppressing calcitriol levels by increasing dietary calcium is an attractive target for obesity intervention. In support of this concept, transgenic mice expressing the agouti gene specifically in adipocytes (a human-like pattern) respond to low-calcium diets with accelerated weight gain and fat accretion, whereas high-calcium diets markedly inhibit lipogenesis, accelerate lipolysis, increase thermogenesis, and suppress fat accretion and weight gain in animals maintained at identical caloric intakes. Further, low-calcium diets impede body fat loss, whereas high-calcium diets markedly accelerate fat loss in transgenic mice subjected to caloric restriction. Dairy sources of calcium exert markedly greater effects in attenuating weight and fat gain and accelerating fat loss. This augmented effect of dairy products is likely due to additional bioactive compounds in dairy that act synergistically with calcium to attenuate adiposity. These concepts are confirmed by both epidemiological and clinical data, which demonstrate that increasing dietary calcium results in significant reductions in adipose tissue mass in obese humans in the absence of caloric restriction and markedly accelerates the weight and body fat loss secondary to caloric restriction, whereas dairy products exert significantly greater effects. These data indicate an important role for dairy products in both the prevention and treatment of obesity. PMID- 12733748 TI - In vitro comparison of hepatic metabolism of 9cis-11 trans and 10trans-12cis isomers of CLA in the rat. AB - Hepatic metabolism of the two main isomers of CLA (9cis-11trans, 10trans-12cis C18:2) was compared to that of oleic acid (representative of the main plasma FA) in 16 rats by using the in vitro method of incubated liver slices. Liver tissue samples were incubated at 37 degrees C for 17 h under an atmosphere of 95% O2/5% CO2 in a medium supplemented with 0.75 mM of FA mixture (representative of circulating nonesterified FA) and with 55 microM [1-(14)C]9cis-11trans C18:2,11 (14)C]10trans-12cis C18:2, or 11-(14)C]oleate. The uptake of CLA by hepatocytes was similar for both isomers (9%) and was three times higher (P < 0.01) than for oleate (2.6%). The rate of CLA isomer oxidation was two times higher (49 and 40% of incorporated amounts of 9cis-11 trans and 10trans-12cis, respectively) than that of oleate (P < 0.01). Total oxidation of oleate and CLA isomers into [14CO2] was low (2 to 7% of total oxidized FA) compared to the partial oxidation (93 to 98%) leading to the production of [14C] acid-soluble products. CLA isomers escaping from catabolism were both highly desaturated (26.7 and 26.8%) into conjugated 18:3. Oleate and CLA isomers were mainly esterified into neutral lipids (70% of esterifled FA) and, to a lesser extent, into polar lipids (30%). They were slowly secreted as parts of VLDL particles (< 0.4% of FA incorporated into cells), the extent of secretion of oleate and of 10trans-12cis being 2.2 fold higher than that of 9cis-11trans (P < 0.02). In conclusion, this study clearly showed that both CLA isomers were highly catabolized by hepatocytes, reducing their availability for peripheral tissues. Moreover, more than 25% of CLA escaping from catabolism was converted into conjugated 18:3, the biological properties of which remain to be elucidated. PMID- 12733747 TI - Treatment and prevention of obesity: what works, what doesn't work, and what might work. AB - We provide a very broad conceptual overview of some of the issues involved in the treatment and prevention of obesity. Data suggest that clinicians have some ability to promote positive changes with obesity treatment. The environment, though important in influencing one's degree of adiposity, has largely transient effects that do not tend to carry over from one time to substantially later times. In contrast, the genetic influences on body mass index at any one time do tend to carry over to later times. This information influences the types of approaches that are and are not likely to be successful in terms of preventing obesity or reducing obesity on a population level. A second issue concerns the composition of weight lost. Conditional on fat loss, weight loss has been associated with an increased mortality rate (MR) whereas, conditional upon weight loss, fat loss has been associated with a decreased MR. This suggests that we should seek treatments that maximize the proportion of weight lost as fat. Third, the efficacy of current treatments is far below patients' expectations and desires. We need both to increase the efficacy of our treatments dramatically and help patients adjust their expectations so that they can take satisfaction in smaller weight losses. Perhaps, with continued efforts at enhancing treatments, we will see incremental advances in the treatment and prevention of obesity. PMID- 12733749 TI - Effects of policosanols and phytosterols on lipid levels and cholesterol biosynthesis in hamsters. AB - The current study was carried out to examine the effects of policosanols and phytosterols, alone and in combination, on lipid profiles, cholesterol biosynthesis, and tissue histopathological changes in hamsters. Fifty male Golden Syrian hamsters, weighing 100 to 120 g, were fed a regular rodent chow for 2 wk before being randomly assigned into 5 groups of 10 animals each fed semisynthetic diets for 4 wk. Group 1 was given a control diet that contained 0.25% cholesterol and 5% fat with a PUFA to saturated FA ratio of 0.4. Groups 2 to 5 were fed the control diet and given Octa-6 [a policosanol mixture from sugar cane wax, 25 mg/kg body weight (BW)], Ricewax (a policosanol mixture from rice wax with 50% being converted to the corresponding acids, 50 mg/kg BW), phytosterols (Cholestatin; 1,000 mg/kg BW), and Ricewax (50 mg/kg BW) plus phytosterols (1,000 mg/kg BW), respectively. The results showed that there was no difference between Octa-6 and Ricewax treatments in any of the lipid parameters measured, and both had similar levels of triglyceride (TG), total cholesterol (T-C), and HDL cholesterol (HDL-C) as the control. Octa-6 but not Ricewax increased (P = 0.03) non-HDL-C as compared with the control. Phytosterols reduced T-C (P < 0.0003) and HDL-C (P < 0.004) without a significant effect on TG and non-HDL-C as compared to the control. Ricewax plus phytosterols had effects similar to those with phytosterols alone. Free cholesterol synthetic rates were not different among the treatments. Policosanols or phytosterols did not show any toxic effects in liver, heart, brain, or kidney. Results suggest that, although phytosterols reduce T-C and HDL-C levels, policosanols have no significant favorable effect in changing lipid levels in hamsters. PMID- 12733750 TI - Effects of high-gamma-linolenic acid canola oil compared with borage oil on reproduction, growth, and brain and behavioral development in mice. AB - Previous research in rats and mice has suggested that gamma-linolenic acid (GLA) derived from borage oil (BO: 23% GLA) may be an appropriate source for increasing levels of long-chain n-6 FA in the developing brain. Recently, transgenic technology has made available a highly enriched GLA seed oil from the canola plant (HGCO: 36% GLA). The first objective of this study was to compare the effects of diets containing equal levels of GLA (23%) from either BO or HGCO on reproduction, pup development, and pup brain FA composition in mice. The second objective was to compare the effects of the HGCO diluted to 23% GLA (GLA-23) with those of undiluted HGCO containing 36% GLA (GLA-36). The diets were fed to the dams prior to conception and throughout pregnancy and lactation, as well as to the pups after weaning. The behavioral development of the pups was measured 12 d after birth, and anxiety in the adult male offspring was assessed using the plus maze. The findings show that despite equivalent levels of GLA, GLA-23 differed from BO in that it reduced pup body weight and was associated with a slight increase in neonatal pup attrition. However, there were no significant effects on pup behavioral development or on performance in the plus maze. An increase in dietary GLA resulted in an increase in brain 20:4n-6 and 22:4n-6, with a corresponding decrease in 22:6n-3. Again, despite their similar levels of GLA, these effects tended to be larger in GLA-23 than in BO. In comparison with GLA 23, GLA-36 had larger effects on growth and brain FA composition but no differences with respect to effects on reproduction and behavioral development. These findings suggest that the HGCO can be used as an alternative source of GLA. PMID- 12733751 TI - Natural abundance stable carbon isotope evidence for the routing and de novo synthesis of bone FA and cholesterol. AB - This research reported in this paper investigated the relationship between diet and bone FA and cholesterol in rats raised on a variety of isotopically controlled diets comprising 20% C3 or C4 protein (casein) and C3 and/or C4 nonprotein or energy (sucrose, starch, and oil) macronutrients. Compound-specific stable carbon isotope analysis (delta13C) was performed on the FA (16:0, 18:0, 18:1, and 18:2) and cholesterol isolated from the diet (n = 4) and bone (n = 8) of these animals. The dietary signals reflected by the bone lipids were investigated using linear regression analysis. delta13C values of bone cholesterol and stearic (18:0) acid were shown to reflect whole-diet delta13C values, whereas the delta13C values of bone palmitic (16:0), oleic (18:1), and linoleic (18:2) acids reflected dietary FA delta13C values. Dietary signal differences are a result of the balance between direct incorporation (or routing) and de novo synthesis of each of these bone lipids. Estimates of the degree of routing of these bone lipids gleaned from correlations between delta13C(dlipid wdiet) (= delta13C(diet lipid) - delta13C(whole diet)) spacings and delta13C(blipid-wdiet) (= delta13C(bone lipid) - delta13C(whole diet)) fractionations demonstrated that the extent of routing, where 18:2 > 16:0 > 18:1 > 18:0 > cholesterol, reflected the relative abundances of these lipids in the diet. These findings provide the basis for more accurate insights into diet when the delta13C analysis of bone fatty FA or cholesterol is employed. PMID- 12733752 TI - Whole-body utilization of n-3 PUFA in n-6 PUFA-deficient rats. AB - We evaluated the utilization of a-linolenic acid (18:3n-3) in growing rats consuming a diet deficient in n-6 PUFA. After 90 d, whole-body 18:3n-3 accumulation was 55% lower, total n-3 PUFA accumulation was 21% lower, and 18:3n 3 disappearance was 14% higher in n-6 PUFA-deficient rats. Part of the reduction of whole-body 18:3n-3 in n-6 PUFA-deficient rats was due to the 25% increase in net conversion of 18:3n-3 to long-chain n-3 PUFA. Despite adequate 18:3n-3 intake, n-6 PUFA deficiency decreased the accumulation of 18:3n-3 and total n-3 PUFA. PMID- 12733753 TI - The historical trauma response among natives and its relationship with substance abuse: a Lakota illustration. AB - Historical trauma (HT) is cumulative emotional and psychological wounding over the lifespan and across generations, emanating from massive group trauma experiences; the historical trauma response (HTR) is the constellation of features in reaction to this trauma. The HTR often includes depression, self destructive behavior, suicidal thoughts and gestures, anxiety, low self-esteem, anger, and difficulty recognizing and expressing emotions. It may include substance abuse, often an attempt to avoid painful feelings through self medication. Historical unresolved grief is the associated affect that accompanies HTR; this grief may be considered fixated, impaired, delayed, and/or disenfranchised. This article will explain HT theory and the HTR, delineate the features of the HTR and its grounding in the literature, offer specific Native examples of HT and HTR, and will suggest ways to incorporate HT theory in treatment, research and evaluation. The article will conclude with implications for all massively traumatized populations. PMID- 12733755 TI - Community readiness: the journey to community healing. AB - Community readiness is a research-based theory that provides a basic understanding of the intervention process in communities. This theory allows us to accurately describe the developmental level of a community relative to a specific issue or problem. In order to move the community toward implementing and maintaining efforts that are effective and sustainable, community mobilization must be based on involvement of multiple systems and utilization of within community resources and strengths. Successful local prevention and intervention efforts must be conceived from models that are community-specific, culturally relevant, and consistent with the level of readiness of the community to implement an intervention. The community readiness model is an innovative method for assessing the level of readiness of a community to develop and implement prevention programming. It can be used as both a research tool to assess distribution of levels of readiness across a group of communities or as a tool to guide prevention efforts at the individual level. This tool has proven useful in addressing a gamut of problems ranging from health and nutritional issues to environmental and social issues. The model identifies specific characteristics related to different levels of problem awareness and readiness for change. PMID- 12733754 TI - Healthy nations: reducing substance abuse in American Indian and Alaska Native communities. AB - Since 1993, 14 American Indian and Alaska Native (AIAN) communities have worked diligently to reduce the harm due to substance abuse in their communities. Funded by the Robert Wood Johnson Foundation's Healthy Nations Initiative I, these communities implemented creative strategies that span the continuum from community-wide prevention, early identification and treatment to aftercare. Drawing upon the unique strengths of their own cultural traditions to find solutions to local substance abuse problems, these efforts have identified important and useful lessons for not only other AIAN communities, but also for sponsors of substance abuse programming in Indian country and elsewhere. Described here are successful strategies for developing and sustaining substance abuse programs in AIAN communities and an assessment of their impacts and accomplishments. PMID- 12733756 TI - Sweat lodge ceremonies for jail-based treatment. AB - Sweat lodge ceremonies (SLCs) have been an integral part of Navajo culture for hundreds of year. The Dine' Center for Substance Abuse Treatment staff utilized SLCs as a modality for jail-based treatment. Data were collected from the Spring of 1996 through the Spring of 1999 from 190 men ranging in age from 18 to 64. These inmate/patients (IPs) provided information at intake on a broad range of questions which were important in understanding the problems these men were having with alcohol and other drugs. Experiential data were collected from 123 IPs after each SLC. Several cultural variables showed improvement in the IP's world view following the SLCs. Even though there were few areas where data were statistically significant, several drinking measures changed in a positive direction. For example, among those subjects who were followed-up, analysis revealed a decrease in the number of drinks consumed in drinking sessions from a mean of 6.7 drinks at intake to a mean of 5.3 drinks. This article examines the role of SLCs in traditional counseling in jail-based treatment of alcohol abuse. PMID- 12733757 TI - A holistic system of care for Native Americans in an urban environment. AB - The Native American Health Center has implemented a holistic system of care in the San Francisco Bay Area as a result of a multiyear strategic planning process that included a needs assessment based on the community-readiness model. The strategic plan links substance abuse, mental health, HIV/AIDS, and social services in a holistic approach congruent with Native American values and traditions. The plan also links prevention with treatment in a continuum of care. Based on a collaboration of Native American nonprofit community-based organizations and public agencies, the plan has resulted in bringing significant resources to the community. PMID- 12733758 TI - Cultural connection and transformation: substance abuse treatment at Friendship House. AB - This research investigates the transformational experiences of Native Americans during the course of their retraditionalization and treatment for drug and alcohol dependence. Twelve graduates of the residential treatment program at Friendship House in San Francisco were interviewed about the kind and quality of their healing experiences in this particular program which provides a comprehensive matrix of Native American medicine as well as Western models of psychological treatment. The purpose of this study was to understand and document the experience of substance abuse recovery from the perspective of the Native Americans in treatment. Twelve themes emerged from the data analysis: feeling cared for; spiritual experience; insight; making a commitment; empowerment/self esteem; releasing emotional pain; remorse; reconnecting to traditional values; forgiveness; relief; safety; and gratitude. PMID- 12733759 TI - The Women's Circle comes full circle. AB - Women have been the backbone of service provision for health and healing in the Native American community in the San Francisco Bay Area. The contributions of Native women are exemplified in the Women's Circle of the Native American Health Centers in San Francisco and Oakland. Women receive a broad range of services through the Women's Circle--in a coed residential substance abuse treatment facility (Friendship House), in groups, in one-on-one counseling, and at the Friendship House American Indian Lodge, a residential women and children's facility in Oakland. This article will look at lessons learned, using both quantitative outcome measures and ethnographic means to examine the impact of the Women's Circle and how the circle was completed. Program staff were interviewed to gain insight into how the program impacted female clients. The article juxtaposes the program elements clients identified as important to their healing and staff's perceptions about the growth of the program. Distinct women's health issues--physical, emotional, mental and spiritual-sculpted program development. Significant findings and lifestyle changes that occurred around involvement in the Women's Circle are examined. PMID- 12733760 TI - Substance dependency among homeless American Indians. AB - Extensive qualitative research in the San Francisco Bay Area in California and in Tucson, Arizona, indicates strong associations between substance abuse and homelessness among American Indians. This article takes a comparative approach to describe and analyze precipitating factors and survival patterns of those who are both homeless and who suffer from substance dependency. Possible precipitating factors presented through case studies consider the complex interaction of childhood fostering or adoption into non-Native families, different types of involuntary institutionalization during youth, and the personal impact of accident, trauma and loss. Coping strategies and keys to survival are examined, including the role of the extended family and close friendships, American Indian and mainstream organizations that offer formal and informal services, the existence of anchor or key households, the helping relationships and sobriety groups among homeless individuals, spirituality, and cultural resiliency. PMID- 12733761 TI - Substance abuse treatment outcomes among American Indians in the Telephone Aftercare Project. AB - Thirty American Indians who had successfully completed their residential substance abuse treatment program with Phoenix-based Native American Connections, Inc. agreed to participate in the Telephone Aftercare Program when they returned to their homes on the reservation. They were provided with graduated monthly aftercare contacts through the telephone for a nominal period of six months, during which time they were interviewed monthly regarding their substance use, and on other life domains at three and six months following discharge from their residential program. Using their baseline data as the comparison, results suggest that six months following residential treatment, clients showed decreased drinking and other drug use, had fewer encounters with the criminal justice system, and had improved familial and social interactions and relationships. PMID- 12733762 TI - Gender differences and conduct disorder among American Indian adolescents in substance abuse treatment. AB - Studies have found high prevalences of conduct disorder among youth in residential substance abuse treatment programs, but no studies have examined this issue among American Indian adolescents while taking into account gender differences. The Diagnostic Interview Schedule for Children, Youth Version and the Composite International Diagnostic Interview, Substance Abuse Module were used to determine the diagnostic status of 89 American Indian adolescents aged between 13 and 18 years who were admitted to a residential treatment program. Seventy-four percent of the adolescents met full DSM-IV criteria for conduct disorder. More than 85% of the participants met at least one criterion for conduct disorder. Conduct disorder was common among both boys and girls in this sample of American Indian adolescents, but the specific antisocial behaviors displayed and their relationships to other psychiatric disorders varied by gender. PMID- 12733763 TI - Incorporating Yup'ik and Cup'ik Eskimo traditions into behavioral health treatment. AB - The Yup'ik and Cup'ik Eskimo of Southwest Alaska have developed programs to provide healing from historical trauma. The Village Sobriety Project, a Center for Substance Abuse Treatment-funded project administered by the Yukon-Kuskokwim Health Corporation from 1999 to 2002, is an effort to incorporate the local Yup'ik and Cup'ik peoples' ways into treatment services for mental health and substance abuse. Activities such as tundra walks and time with elders are supported in treatment, and Medicaid reimbursement was successfully obtained for such activities. Documentation of treatment showed that it incorporated cultural elements of the Yup'ik and Cup'ik people. PMID- 12733764 TI - Mobilizing communities to reduce substance abuse in Indian country. AB - The impact of substance abuse is notable on Indian reservations in the United States and in the border communities surrounding them. One Indian country community, McKinley County, New Mexico, developed and implemented a series of alcohol policy and programmatic reforms beginning in 1989 which have reduced the impact of substance abuse on this community. Learning from the McKinley County experience, Fremont County in Wyoming, home of the Wind River Indian Reservation, has implemented similar reforms. This article introduces the substance abuse reduction efforts and delineates specific innovations implemented in these communities. The influence that these two communities have had on changes in substance abuse policy and programming statewide in New Mexico and Wyoming is also reviewed. PMID- 12733765 TI - Duck and cover: a prudent defense against smallpox. PMID- 12733766 TI - Medicaid reimbursement. PMID- 12733767 TI - Trailblazers overlooked? PMID- 12733768 TI - Caries into dentin. PMID- 12733769 TI - Caring for special patients. PMID- 12733770 TI - Caring for special patients. PMID- 12733772 TI - Caring for special patients. PMID- 12733771 TI - Caring for special patients. PMID- 12733773 TI - ADA Foundation streamlines, enhances association charitable activities. PMID- 12733774 TI - Periodontal disease linked with Kindler syndrome. PMID- 12733775 TI - Does keeping up with the latest technology affect the quality of patient care? PMID- 12733776 TI - Vertical root fractures: clinical and radiographic diagnosis. AB - BACKGROUND: Early detection and management of vertical root fractures, or VRFs, remain a vexing issue that has caused needless suffering for patients as well as for dentists. The authors present techniques to aid the dentist in recognizing VRFs. METHODS: During a five-year period, the authors examined 36 patients who had VRFs. Absent control subjects and a larger number of patients, the authors did not design this investigation for statistical analysis. They diagnosed VRFs through dental histories and clinical and radiographic examinations. RESULTS: The study revealed VRFs in 36 teeth, two of which were vital and 34 of which were nonvital (that is, endodontically treated). The 34 VRFs resulted from excessive operative procedures performed in the root canal after endodontic therapy. Thirty one of these 34 VRFs were caused by poorly designed dowels (too long, too wide or both) or inappropriate selection of the tooth as a bridge abutment; two VRFs were caused by a restoration that exerted lateral pressure on the axial walls of the preparation; and one VRF was caused by overzealous endodontic forces. The VRFs in the two vital teeth were in men who had a history of bruxism or clenching. CONCLUSIONS AND CLINICAL IMPLICATIONS: VRFs can be detected early by listening to the patient's chief complaints, carefully examining periapical and bitewing radiographs and performing a thorough clinical examination. PMID- 12733777 TI - The role of oral appliances in treating obstructive sleep apnea. AB - BACKGROUND: Sleep-related breathing disorders are common and often are associated with vascular complications such as arterial hypertension, coronary heart disease and stroke. The most widely studied form of these disorders is obstructive sleep apnea. Patients usually are diagnosed with obstructive sleep apnea years after the onset of symptoms, which generally are nonspecific and include excessive daytime sleepiness, chronic fatigue and habitual snoring. The risk factors for sleep apnea are obesity, advancing age, male sex and maxillofacial abnormalities. This review focuses on the diagnosis and treatment of obstructive sleep apnea and, specifically, on the utility of oral appliances in the management of this disorder. METHODS: The review is based on a MEDLINE search for articles in English on this topic. The article discusses results of randomized studies and prospective case series. CLINICAL IMPLICATIONS: Several treatment options are quite effective. Nasal continuous positive airway pressure, with an overall acceptance rate of 70 percent, is the most widely used treatment modality. Maxillofacial surgery, although effective, is reserved for patients who have not responded to the more conventional therapies. Newer methods include application of oral appliances. Oral appliances have been shown to alleviate the severity of respiratory disturbances during sleep by about 60 percent, with an overall acceptance rate of 75 percent. The long-term complications generally are minor and are related to occlusal changes and temporomandibular joint discomfort. CONCLUSIONS: With the advent of oral appliances, dentists are increasingly involved in managing the care of patients with sleep-related breathing disorders. Further studies are needed to determine the long-term complications of this type of intervention for the treatment of sleep-related breathing disorders. PMID- 12733778 TI - The unresolved problem of the third molar: would people be better off without it? AB - BACKGROUND: Third molars are teeth that have little functional value and a relatively high rate of associated pain and disease. Their value as part of the dentition of modern people is dubious. TYPES OF STUDIES REVIEWED: The authors review the evolution, development, morbidity and treatment of third molars. They assess the value of third molars in the 21st century and describe the risks these teeth pose when they develop in the dentition. CONCLUSIONS: There is a mandate for the dental profession to improve health outcomes and quality of life. The prevention of third molar-related morbidity should be included in dental research efforts. The authors suggest that novel preventive methodologies be developed to alleviate the problems third molars pose. One potential methodology suggested is intentional therapeutic agenesis of this tooth. CLINICAL IMPLICATIONS: Prevention of third molar development early in life, even before tooth bud initiation, could dramatically improve health care outcomes for millions of people. PMID- 12733779 TI - Microarrays and clinical dentistry. AB - BACKGROUND: The Human Genome Project, or HGP, has inspired a great deal of exciting biology recently by enabling the development of new technologies that will be essential for understanding the different types of abnormalities in diseases related to the oral cavity. LITERATURE REVIEWED: The authors review current literature pertaining to the advanced microarray technologies arising from the HGP and how they can contribute to dentistry. This technology has become a standard tool for monitoring activities of genes at both academic and pharmaceutical research institutions. RESULTS: With the availability of the DNA sequences for the entire human genome, attention now is focused on understanding various diseases at the genome level. Deciphering the molecular behavior of genetically encoded proteins is crucial to obtaining a more comprehensive picture of disease processes. Important progress has been made using microarrays, which have been shown to be effective in identifying gene expression patterns and variations that correlate with cellular development, physiology and function. Arrays can be used to classify tissue samples accurately based on molecular profiles and to select candidate genes related to a number of cancers, including oral cancer. This type of oral genetic approach will aid in the understanding of disease progression, thus improving diagnosis and treatment for patients. CLINICAL IMPLICATIONS: Microarrays hold much promise for the analysis of diseases in the oral cavity. As the technology evolves, dentists may see these tools as screening tests for better managing patients' dental care. PMID- 12733781 TI - Fabrication of immediate transitional denture for patients with fixed partial dentures. PMID- 12733780 TI - Direct and indirect restorative materials. AB - BACKGROUND: In recent years, dentistry has benefited from a marked increase in the development of esthetic materials, including ceramic and plastic compounds. But the advent of these new materials has not eliminated the usefulness of more traditional restorative materials such as gold, base metal alloys and dental amalgam. OVERVIEW: This report outlines important features of direct and indirect restoratives, with an emphasis on the safety and efficacy of each material. CONCLUSIONS AND PRACTICE IMPLICATIONS: This article was developed to help dentists explain to their patients the relative pros and cons of various materials used in dental restorations, which include fillings, crowns, bridges and inlays. The weight of the scientific evidence indicates that all of these materials are safe and effective for their intended use. Patients, in consultation with their dentists, are free to choose the most appropriate among them for their particular needs. PMID- 12733782 TI - Proper use of beryllium-containing alloys. AB - BACKGROUND: To assist dentists in providing a safe and healthful workplace, the ADA Council on Scientific Affairs developed this advisory statement on the proper use of beryllium-containing alloys in the dental operatory and laboratory. OVERVIEW: Recommendations are made for reducing the potential risk of beryllium exposure for employees working in dental operatories or laboratories who fabricate or manipulate beryllium-containing alloys in dental prostheses. CONCLUSIONS AND PRACTICE IMPLICATIONS: The Council recommends that practitioners adhere to the Ocupational Safety and Health Administration's recommendations for reducing beryllium exposure in the dental operatory and laboratory. These include the recommendation that--where possible--practitioners use alloys that do not contain beryllium in the fabrication of dental prostheses. PMID- 12733783 TI - Oral health status of older rural adults in the United States. AB - BACKGROUND: Little is known about the oral health care of older rural residents. The authors describe oral health indicators for the older adult population by place of residence in the United States. METHODS: The authors analyzed data from the Third National Health and Nutrition Examination Survey and the 1995, 1997 and 1998 National Health Interview Surveys. Oral health indicators included perceived oral health (self-reported dental status and unmet dental needs) and dental status (untreated caries; decayed, missing and filled permanent teeth, or DMFT; and edentulism). Dental care utilization and access were measured by number of dental visits, frequency of dental visits and dental insurance status. RESULTS: Older rural adults were more likely than their urban counterparts to be uninsured for dental care (72.1 percent versus 66.1 percent, respectively) and were less likely to report dental visits in the past year (46.9 percent versus 58.4 percent, respectively). A higher proportion of rural residents than urban residents were edentulous (36.7 percent versus 28.2 percent, respectively) and reported poor dental status (50.7 percent versus 42.2 percent, respectively). There were no differences in unmet dental needs, percentage of people with untreated caries or in mean DMFT by place of residence. CONCLUSIONS: Older rural residents inadequately utilize dental care and have less favorable oral health indicators than do older urban residents. CLINICAL IMPLICATIONS: This article shows the need for more dental practitioners in rural areas. With the low density of dentists per person and the high need for care, rural America offers an excellent opportunity for oral health professionals to provide much needed services. PMID- 12733784 TI - Intraosseous anesthesia: implications, instrumentation and techniques. AB - BACKGROUND: The author reviews historical methods and the instruments used to bring about intraosseous anesthesia, or IOA; discusses the criteria for successful use of the intraosseous injection, or IOI, technique; and provides recommendations. TYPES OF STUDIES REVIEWED: Articles from before 1990 consisted of subjective reports of patient types and procedures performed using IOI as a primary technique. Studies published after 1990 yielded subjective findings on indications for expanded clinical use. The author discusses the expansion of the role of IOI relative to integrated local anesthetic delivery systems. RESULTS: The literature and studies verify the efficacy of IOI as a supplemental or primary technique. The author recommends anesthetics and infusion sites, and reports on the patients' perceptions of comfort. CONCLUSIONS AND CLINICAL IMPLICATIONS: IOI can be used as a supplemental or primary technique to bring about local anesthesia in routine dental procedures. It can be used as a supplemental technique with mandibular nerve blocks to enhance deep pulpal anesthesia. It can be used as a primary technique so that patients do not experience numb lips or tongues postoperatively. Dentists can appreciate the immediate onset of anesthesia and reduced dosage levels of anesthetics associated with using IOI. PMID- 12733786 TI - Louisiana dental advertising regulations upheld. PMID- 12733785 TI - Applying statistics to the dental practice. AB - There are numerous variables in the specific mix of procedures a practice performs. Other practice production variables include the number of patients a practice serves and the number of team members available to accomplish the practice objectives. The majority of practices would benefit by following a model similar to the one above. To return to the McDonald's example, success is measured according to whether each McDonald's sells the optimal product mix for its customers. It doesn't matter what components make up that mix. The same thing is generally true for dental practices. Conducting a "percentage production analysis" to determine how closely a practice follows the above model can be beneficial. It can help in making decisions regarding what changes need to be made to achieve optimal production levels and greater practice success. PMID- 12733787 TI - Coping with the changing state of dental managed care. AB - About 58 percent of the American population has some type of managed dental care that pays for some, not all, dental care needs. Unlike the medical profession, the dental profession has not been devoured by managed care, but there is no question that managed care plans are here to stay. The various types of managed care programs are changing every day. The trend is for DHMOs and dental indemnity programs to decrease, DPPOs and dental referral programs to increase and dental reimbursement programs to grow slowly as more organizations become aware of their benefits. Plans vary from excellent to poor in their service to patients and their dentists. We dentists must learn to live with them and to educate our patients about the advantages and disadvantages of each. I am pleased to observe that fee-for-service dentistry, although challenged, is still very alive and strong and serving the public with the same high quality and freedom of choice for which American dentistry has always been known. PMID- 12733789 TI - For the dental patient. Do you have a cracked tooth? PMID- 12733788 TI - Resin-based composites. PMID- 12733790 TI - Giardia and Cryptosporidium in pristine protected catchments in central eastern Australia. AB - We report the first confirmed records of the pathogenic protozoa Giardia lamblia and Cryptosporidium parvum from small remote streams in pristine protected areas in Australia, beyond the reach of urban or agricultural contamination. PMID- 12733791 TI - Hypersaline cyanobacterial mats as indicators of elevated tropical hurricane activity and associated climate change. AB - The Atlantic hurricanes of 1999 caused widespread environmental damage throughout the Caribbean and US mid-Atlantic coastal regions. However, these storms also proved beneficial to certain microbial habitats; specifically, cyanobacteria dominated mats. Modern mats represent the oldest known biological communities on earth, stromatolites. Contemporary mats are dominant biological communities in the hypersaline Bahamian lakes along the Atlantic hurricane track. We examined the impacts of varying levels of hypersalinity on 2 processes controlling mat growth, photosynthesis and nitrogen fixation, in Salt Pond, San Salvador Island, Bahamas. Hypersalinity (> 5 times seawater salinity) proved highly inhibitory to these processes. Freshwater input from Hurricane Floyd and other large storms alleviated this salt-inhibition. A predicted 10 to 40 year increase in Atlantic hurricane activity accompanied by more frequent "freshening" events will enhance mat productivity, CO2 sequestration and nutrient cycling. Cyanobacterial mats are sensitive short- and long-term indicators of climatic and ecological changes impacting these and other waterstressed environments. PMID- 12733792 TI - Regional differences in the occurrence of understorey species reflect nitrogen deposition in Swedish forests. AB - Possible links between the occurrence of Vaccinium myrtillus, V. vitis-idaea and Deschampsia flexuosa and rates of nitrogen deposition were investigated in 557 coniferous forest stands. In areas with high N-deposition, V. myrtillus was less frequent, less abundant and more susceptible to the leaf pathogen Valdensia heterodoxa than in areas with lower levels of N-deposition. The occurrence of V. vitis-idaea was also strongly negatively correlated with increasing N-deposition, but no such trend was found for D. flexuosa. In regions with high N-deposition, V. myrtillus was more common in stands dominated by Scots pine than in stands dominated by Norway spruce. This was not the case in regions with lower levels of N-deposition. The patterns observed accord with results from N-addition experiments that demonstrate significant effects on vegetation, caused by N deposition. The data suggest that even low rates of N-deposition may decrease the abundance of the most dominant species in coniferous forest ground flora. PMID- 12733793 TI - Fish status survey of Nordic lakes: effects of acidification, eutrophication and stocking activity on present fish species composition. AB - The status of fish populations in 3821 lakes in Norway, Sweden and Finland was assessed in 1995-1997. The survey lakes were chosen by stratified random sampling from all (126 482) Fennoscandian lakes > or = 0.04 km2. The water chemistry of the lakes was analyzed and information on fish status was obtained by a postal inquiry. Fish population losses were most frequent in the most highly acidified region of southern Norway and least common in eastern Fennoscandia. According to the inquiry results, the number of lost stocks of brown trout (Salmo trutta), roach (Rutilus rutilus), Arctic char (Salvelinus alpinus) and perch (Perca fluviatilis) was estimated to exceed 10000. The number of stocks of these species potentially affected by the low alkalinity of lake water was estimated to exceed 11000. About 3300 lakes showed high total phosphorus (> 25 microg L(-1)) and cyprinid dominance in eastern Fennoscandia, notably southwestern Finland. This survey did not reveal any extinction of fish species due to eutrophication. One third of the lakes had been artificially stocked with at least one new species, most often brown trout, whitefish (Coregonus lavaretus s.l.), Arctic char, rainbow trout (Oncorhynchus mykiss), pike-perch (Stizostedion lucioperca), grayling (Thymallus thymallus), pike (Esox lucius), bream (Abramis brama), tench (Tinca tinca) and European minnow (Phoxinus phoxinus). The number of artificially manipulated stocks of these species in Fennoscandian lakes was estimated to exceed 52000. Hence, the number of fish species occurring in Nordic lakes has recently been changed more by stockings than by losses of fish species through environmental changes such as acidification. PMID- 12733794 TI - Assessment of management-dependent nutrient losses in tropical industrial tree plantations. AB - Industrial tree plantations in the tropics usually follow short rotations and intensive site management including slash and burn, and the use of heavy machinery. We attempt to quantify the implied nutrient losses (harvest export, erosion, slash and burn, leaching) in order to give plantation managers a chance to understand the significance of their planning and decisions. We used the scarce globally available information and a case study plantation in East Kalimantan, Indonesia (Acacia mangium and Eucalyptus deglupta). Adaptation involves problems and is discussed in some detail. Results are approximate only. Assuming a harvest volume of 200 m3 ha(-1), we assessed a loss of 427-680 kg ha( 1) N, 12-13 kg ha(-1) P, 178-252 kg ha(-1) Ca, 276-370 kg ha(-1) K, and 45-57 kg ha(-1) Mg per rotation. Of this overall loss, stand harvest accounted for 18-29% (N), 21-30% (P), 56-26% (K), 48-64% (Ca) and 22-37% (Mg). This means that the cumulative loss by erosion, slash and burn, and leaching exceeds that of the harvest. These losses can be influenced by management. PMID- 12733795 TI - Pastoral ecosystems and the issue of scale. AB - This paper uses examples from Kenya and the Sudan to argue that the scale at which we pitch our analysis when trying to identify the basic properties of pastoral ecosystems may not be appropriate when it comes to recommending policy measures to secure the continued viability of pastoral herding. Pastoral households are always parts of large-scale economic and social structures. In many cases, such integration has provided the basis for the continued viability of pastoral adaptations. In other cases, the changing nature of local economies and social relations following from integration into large systems, has threatened the viability of pastoral herding or led to increasing differentiation within and between local communities. This paper raises the issue of how we most fruitfully define and delimit ecological and social systems in different local settings. It also questions conventional approaches to community-based natural resource management, which are now attracting widespread international attention. PMID- 12733796 TI - Sustainability appraisal of shifting cultivation in the Chittagong Hill Tracts of Bangladesh. AB - An integrated socioeconomic and erosion study on the sustainability of traditional shifting cultivation (Jhum) carried out in 1998 and 1999 in the Chittagong Hill Tracts (CHT) of Bangladesh showed the system to be nonsustainable under the current conditions with fallow periods of only 3-5 years and lack of land rights. An estimated input (mainly labor) of USD 380 ha(-1) yr(-1) results in only a total output of USD 360 ha(-1) yr(-1) and Jhum cultivated areas suffer severe loss of soil and valuable plant nutrients along with runoff during the rainy season. To compensate these losses by commercial fertilizers will cost nearly USD 2 million yr(-1) for CHT. To ensure long-term productivity of the soils, Jhum should therefore be adjusted to a tolerable level and farmers should be given rights and title to the land to motivate them to switch to improved, settled farming systems. PMID- 12733797 TI - Water management for a megacity: Mexico City Metropolitan Area. AB - The paper presents an overview of the present situation of the Mexico City Metropolitan Area (MCMA). The analysis indicates an urgent need to radically improve the current water supply and wastewater management practices, to become sustainable. The MCMA is one of the most rapidly growing urban centers of the world, with a population of about 21 million people, a very high rate of immigration and numerous illegal settlements. In order to meet the increasing water demand, successive governments have focused almost exclusively on supply management and engineering solutions, which have resulted in investments of hundreds of millions of USD and the construction of major infrastructure projects for interbasin water transfer. Environmental, economic and social policies associated with water management are mostly inadequate and insufficient, which is resulting in increasing deterioration in the environment, health and socioeconomic conditions of a population living in one of the largest urban agglomerations of the world. Surprisingly, however, no long-term strategies on demand-management, reuse, conservation, and improved water-management practices have been developed so far. PMID- 12733798 TI - Can corals be harvested sustainably? AB - The international trade in corals has been identified as a potential cause of localized depletion of coral populations in the major coral-exporting countries. The international coral trade is regulated by the Convention on International Trade in Endangered Species of Wild Flora and Fauna (CITES) agreement, which requires that export of corals is not detrimental to the species. The primary coral importing regions (USA and Europe) have threatened to limit or ban coral imports unless sustainable practices can be demonstrated. The spatial and temporal scale at which sustainability is defined is important in evaluating sustainability, e.g. at geological, regional or local scales. Other major issues are: the ecology of the target species; management options including provision of no-take areas; and the potential for coral culture. Implementation of practices that enhance ecological sustainability in the coral harvest fishery is possible, but may be difficult in some developing countries because of limited natural resource management capacity. PMID- 12733799 TI - Reef development at Inhaca Island, Mozambique: coral communities and impacts of the 1999/2000 Southern African floods. AB - Inhaca Island, southern Mozambique, is located towards the southerly latitudinal limits of coral reef growth. Reef development is restricted to the margins of channels which dissect intertidal flats on the Maputo Bay side of the island, and to depths of around 6 m. In contrast to lower latitude reefs, reef development is therefore both spatially and bathymetrically restricted (largely due to high turbidity levels). These natural stress levels were exacerbated, via increased freshwater and sediment discharge, during the severe floods of late 1999/early 2000. Flood impacts varied but were most significant on reefs on the inner (western) side of the island where live coral cover (LCC) decreased from 60.5% (1999) to 24.0% (2001). This is attributed to freshwater-induced bleaching. Dead in situ coral cover increased from 18.6% (1999) to 51.3% (2001). Reefs on the southern tip of the island, by contrast, were relatively unaffected. It is suggested that this largely reflects a closer proximity to the open Indian Ocean which mitigated the effects of freshwater dilution. PMID- 12733800 TI - Recreational scuba diving in Caribbean marine protected areas: do the users pay? AB - There are more than 200 marine protected areas (MPAs) in the Caribbean and Central America that contain coral reefs and are therefore theoretically attractive to scuba divers. One fifth of dive operators in 30 countries were surveyed for their use of MPAs: the majority are located within 20 km of at least one MPA and 46% conduct at least 80% of their diving within a MPA. An estimated 15 million dives take place outside of Florida each year, half of these occurring inside MPAs. Only 25% of MPAs containing coral reefs charge divers an entry or user fee, which is most usually USD 2-3 levied per dive or per diver. The revenue generated by these fees is estimated at USD 1-2 million annually, but the potential for generating income has not been fully realized. A significant contribution to the cost of regional conservation could be achieved if higher fees were applied more widely than at present. PMID- 12733801 TI - Status and interconnections of selected environmental issues in the global coastal zones. AB - This study focuses on assessing the state of population distribution, land cover distribution, biodiversity hotspots, and protected areas in global coastal zones. The coastal zone is defined as land within 100 km of the coastline. This study attempts to answer such questions as: how crowded are the coastal zones, what is the pattern of land cover distribution in these areas, how much of these areas are designated as protected areas, what is the state of the biodiversity hotspots, and what are the interconnections between people and coastal environment. This study uses globally consistent and comprehensive geospatial datasets based on remote sensing and other sources. The application of Geographic Information System (GIS) layering methods and consistent datasets has made it possible to identify and quantify selected coastal zones environmental issues and their interconnections. It is expected that such information provide a scientific basis for global coastal zones management and assist in policy formulations at the national and international levels. PMID- 12733802 TI - Heavy metals from the River Adayar, India: infiltration into the adjacent groundwater aquifer. AB - In the Adayar riverbed and its estuary 5 undisturbed sediment cores were drilled using a petrol-driven hammer. Cores of a depth of 4-5 m were sampled in 0.5 m steps. The distribution of heavy metals (Cr, Mn, Ni, Cu, Zn, Cd and Pb) in the top layers of fluvial and estuarine sediments indicates recent anthropogenic input: Revealing that the river acts as a sink for drainage from the residential and industrial zones. In this study, the mobilization of heavy metals into greater depths and their probable effects on the groundwater body are discussed. The high concentration of heavy metals and the influent character of the river Adayar allow the mobilization of metal ions and their transport into the deeper layers of the sediment. A changing environment due to effects such as saltwater intrusion and monsoon floods is the driving force for this phenomenon. PMID- 12733803 TI - Fanwort in eastern China: an invasive aquatic plant and potential ecological consequences. PMID- 12733804 TI - Environmental monitoring technology development needs in developing countries. PMID- 12733805 TI - Effects of clay minerals on Cr(VI) reduction by organic compounds. AB - The objective of this study is to examine the effect of clay minerals (illite, montmorillonite, and kaolinite) on chromate (Cr(VI)) reduction by several low molecular weight organic compounds. Batch experiments at pH ranging from 3.0 to 6.0 and 25 degrees C showed that 2:1 layered clays illite and smectite catalyzed Cr(VI) reduction by oxalate. The catalytic effect increased as pH was decreased. The 1:1 clay kaolinite had no catalytic effect under comparable conditions. Direct Cr(VI) reduction by reactive moieties associated with illite and montmorillonite was observed, but at a much slower rate than the catalytic pathway. Cr(VI) reduction by glyoxylic acid, glycolic acid, lactic acid, and mandelic acid was accelerated by illite, although aqueous phase reduction might occur in parallel. These results suggest that Cr(VI) reduction rates in subsurface environments rich in organic compounds may be elevated through catalysis of surface-bound metals and/or soluble species from the clay minerals, and as a result, higher than those expected from aqueous phase reaction alone. Such rate enhancement for Cr(VI) reduction needs to be accounted for when developing new remedial techniques for chromium site remediation or assessing its natural attenuation. PMID- 12733806 TI - Surfactant dissolution and mobilization of LNAPL contaminants in aquifers. AB - Improper disposal, accidental spills and leaks of non-aqueous phase liquids (NAPL) such as gasoline, fuel oil and creosote result in long-term persistent sources of groundwater pollution. Column and 2-D tanks experiments were conducted to evaluate the use of surfactant-enhanced recovery of light non-aqueous phase liquids (LNAPL) in groundwater aquifers. These experiments focused on the use of surfactants to promote dissolution and mobilization in addition to evaluating the increase of aqueous phase permeability as residual NAPL is recovered. Further experiments are presented that show the innovative use of surfactants during primary pumping to recover free product can potentially increase the amount of free product recovered, can potentially reduce the amount of residual NAPL remaining after primary pumping and performs better than the use of surfactants to mobilize trapped residual NAPL. PMID- 12733808 TI - Bioremediation of metal contamination. AB - A study was initiated to evaluate the use of the fungus Aspergillus niger for bioleaching and then to determine the effect of process steps, the tailings concentration and type of substrate. An oxidized mining tailing containing mainly copper (7240 mg kg(-1) tailings) was studied. A sucrose and mineral salts medium was initially used to produce citric and gluconic acids by A. niger at various concentrations of tailings ( 1, 5, 7, 10 and 15% w/v). Maximal removal of up to 60% of the copper was obtained for the 5% tailings when the organic acid supernatant was added to the tailings. In a single step process, A. niger was then grown in the presence of mining tailings at various concentrations. Maximum copper solubilization (63%) occurred with 10% mining tailings using sucrose as the substrate. Other substrates were then evaluated including molasses, corn cobs and brewery waste (10% mining tailings). Sucrose gave the best results for copper removal, followed by molasses, corn cobs and brewery waste. Other experiments using ultrasound as a pretreatment showed that 80% removal of the copper could be obtained for a 5% tailings concentration. In conclusion, leaching of copper from mining tailings is technically feasible using A. niger but further research will be required to increase the economic feasibility of the process. PMID- 12733807 TI - Removal and recovery of metals from a coal pile runoff. AB - The removal and recovery of heavy metals from a coal pile runoff water using a mixture of multiple metal-tolerant bacterial strains of ATCC 55673, and ATCC 55674 and a Pseudomonas sp. was investigated. The analysis of elemental composition of metal precipitates recovered from the bacterial biomass by transmission electron microscopy andenergy dispersive X-ray analysis revealed the presence of metals originally present in the wastewater. In addition, analysis of metals in culture supernatant and bacterial biomass by inductively coupled plasma emission spectroscopy (ICP-ES) indicated a removal range of 82-100% and a recovery of 15-58% of metals from the wastewater and bacterial biomass, respectively. PMID- 12733809 TI - Moisture monitoring in waste disposal surface barriers. AB - Surface barriers for waste disposal sites should prevent waste water and gas emission into the environment. It is necessary to assess their proper operation by monitoring the water regime of the containment. A set of three new water content measuring devices has been developed that provide an economical solution for monitoring the moisture distribution and water dynamic. They will give an early warning service if the barrier system is at risk of being damaged. The cryo soil moisture sensor 'LUMBRICUS' is an in situ self-calibrating absolute water content measuring device. It measures moisture profiles at spot locations down to 2.5 m depth with an accuracy of better than 1.5% and a depth resolution of 0.03 m. The sensor inherently measures density changes and initial cracks of shrinking materials like clay minerals. The large area soil moisture sensor 'TAUPE' is a moisture sensitive electric cable network to be buried in the mineral barrier material of the cover. A report will be given with results and experiences on an exemplary installation at the Waste Disposal Facility Karlsruhe-West. 800 m2 of the barrier construction have been continuously monitored since December 1997. Volumetric water content differences of 1.5% have been detected and localised within 4 m. This device is already installed in two other waste disposal sites. A modified 'TAUPE' was constructed for the control of tunnels and river dams as well. Thin sheet moisture sensor 'FORMI' is specifically designed for moisture measurements in liners like bentonite, textile and plastic. Due to its flexibility it follows the curvature of the liner. The sensor measures independently from neighbouring materials and can be matched to a wide range of different thickness of the material. The sensors are patented in several countries. PMID- 12733810 TI - Biotic landfill cover treatments for mitigating methane emissions. AB - Landfill methane (CH4) emissions have been cited as one of the anthropogenic gas releases that can and should be controlled to reduce global climate change. This article reviews recent research that identifies ways to enhance microbial consumption of the gas in the aerobic portion of a landfill cover. Use of these methods can augment CH4 emission reductions achieved by gas collection or provide a sole means to consume CH4 at small landfills that do not have active gas collection systems. Field studies indicate that high levels of CH4 removal can be achieved by optimizing natural soil microbial processes. Further, during biotic conversion, not all of the CH4 carbon is converted to carbon dioxide (CO2) gas and released to the atmosphere; some of it will be sequestered in microbial biomass. Because biotic covers can employ residuals from other municipal processes, financial benefits can also accrue from avoided costs for residuals disposal. PMID- 12733811 TI - Some engineering aspects of homoionized mixed clay minerals. AB - Many studies have been conducted to investigate the physicochemical behavior of pure clay minerals and predict their engineering performance in the field. In this study, the physicochemical properties of an artificial mixture of different clay minerals namely, 40-50% montmorillonite, 20-30% illite and 10-15% kaolin were investigated. The mixture was homoionized with sodium, Na+; calcium, Ca2+; and aluminum, Al3+. The engineering properties studied were consistency limits, sediment volume, compressibility behavior, and hydraulic conductivity. The results revealed that the liquid, plastic and shrinkage limits of soil increased with increasing cation valence. The hydraulic conductivity of the soil also increased with an increase in the valence of the cation at any given void ratio. Aluminum and sodium treated clays had the highest and the lowest modified compression index values, respectively. Furthermore, trivalent cation saturated clayey soil consolidates three times faster than that of monovalent and two times faster than that of divalent. These properties of the soils determined were, in general, similar to those of kaolinite rather than those of montmorillonite. The comparison of the results obtained with the published data in the literature revealed that the physicochemical behavior of the tested clay soil was, in general, similar to that of kaolinite. PMID- 12733812 TI - Techniques of remote sensing applied to the environmental analysis of part of an aquifer located in the Sao Jose dos Campos Region sp, Brazil. AB - The anthropogenic activity on the surface can modify and introduce new mechanisms of recharging the groundwater system, modifying the tax, the frequency and the quality of recharge of underground waters. The understanding of these mechanisms and the correct evaluation of such modifications are fundamental in determining the vulnerability of groundwater contamination. The groundwater flow of the South Paraiba Compartment, in the region of Sao Jose dos Campos, Sao Paulo, is directly related to structural features of the Taubate Basin and, therefore, the analysis of its behaviour enhances the understanding of tectonic structure. The methodology adopted for this work consists in pre-processing and processing of the satellite images, visual interpretation of HSI products, field work and data integration. The derivation of the main structural features was based on visual analysis of the texture elements of drainage, and the relief in sedimentary and crystalline rocks. Statistical analysis of the feature densities and the metric geometric relations between the analysed elements have been conducted. The crystalline rocks, on which the sediments were laying, conditions and controls the structural arrangement of sedimentary formations. The formation of the South Paraiba Graben is associated with Cenozoic distensive movement which reactivated old features of crust weakness and generated previous cycles with normal characteristics. The environmental analysis is based on the integration of the existing methodology to characterise vulnerability of an universal pollutant and density fracture zone. The digital integration was processed using GIS (Geographic Information System) to delineate five defined vulnerability classes. The hydrogeological settings were analysed in each thematic map and, using fuzzy logic, an index for each different vulnerability class was compiled. Evidence maps could be combined in a series of steps using map algebra. PMID- 12733814 TI - Applying value analysis and fuzzy logic to select areas for installing waste fills. AB - This article discusses a methodology for assessing and ranking a predefined universe of objects to assist in the selection of suitable areas for the construction of sanitary landfills. The methodology is, in principle, for universal application. Its development was initially based on the 'Value Analysis' methodology and then on the 'Fuzzy Eigenvector Method', or 'Fuzzy Logic' as it is sometimes referred to. The focus of the study herein is on the comparative appraisal of a group of elements which are part of the guidelines to be adopted when determining the choice of the most suitable site for a sanitary landfill. This set of guidelines includes technical, economical, environmental, social and political aspects. As an example of the methodology proposed, four different areas were chosen. They were studied to determine which is the most appropriate site for the development of a new solid waste fill site. The results for these four areas, together with the functions chosen and function weights, are presented here. PMID- 12733813 TI - Renewable energy from gasification of manure: an innovative technology in search of fertile policy. AB - After describing an innovative technology, the close-coupled gasification and cyclonic combustor, this article explores the policy issues that inhibit a superior sustainable solution from flourishing. Discussion of technology includes defining biomass, explaining what biomass to energy means, what the advantages of biomass to energy are, and why gasification is a superior biomass to energy technology. Specifically the environmental benefits of alternatives to landspreading of traditional manure management are discussed, as well as the advantages of gasification versus traditional combustion techniques for high nitrogen fuels. The policy environment is explored, particularly regarding sustainability, manure management, and renewable energy. Artificial, non sustainable barriers to renewable energy, and the impact of wide jurisdictional variability are discussed. North Carolina is identified as a unique jurisdiction to monitor because of its high volume of livestock manure, and laggard position in renewable energy advocacy. The authors contend that these two positions are unsustainable, and that pressures can be expected to force the state to modify its renewable energy policies or risk losing market share in livestock production to more pro-sustainable policy oriented states. PMID- 12733816 TI - A quantitative methodology for indexing environmental sensitivity and pollution potential. AB - A methodology for rating the suitability of sites for the location of industrial facilities is formulated and applied to the case of a coal-fired power plant location. The methodology comprises two major interlinked components: the environmental plant location indexing component, which involves the identification, scaling and weighting of environmental sensitivity factors; and the impact analysis component, which involves the superimposition of the pollution generation impacts of an industrial facility on spatially gridded zones of various environmental sensitivities. For each rectangular areal unit defined by a square grid, the Unit Pollution Potential Index is determined by the severity and distribution of key environmental sensitivity factors and the coverage of superimposed pollutant effects as determined by contaminant fate and transport models. For any alternative site of a planned facility, the summation of the unit indices over the area of influence of the facility provides the quantitative Pollution Index, which can be used as a basis for comparison of alternative sites for planned facilities. For this paper, this methodology is applied to the hypothetical case of the siting of a coal-fired power plant in the northeastern region of the United States, in which three alternative sites are considered. The three sites: A, B and C yielded indices of 47.83, 47.91 and 47.6, respectively, indicating that site C is the most suitable for siting the power plant. PMID- 12733817 TI - A technique for cutting brittle undisturbed lateritic soil block samples. AB - This note describes a technique for cutting undisturbed brittle block samples into smaller specimens for further geotechnical testing. This technique revealed very useful in dealing with collapsible soils, where the sampling is recommended to be done with block soil samples. A further use of this technique as an efficient way for sampling collapsible soils is proposed. PMID- 12733815 TI - The combined use of electrokinetic remediation and phytoremediation to decontaminate metal-polluted soils: a laboratory-scale feasibility study. AB - The use of a combination of electrokinetic remediation and phytoremediation to decontaminate two metal-polluted soils has been demonstrated in laboratory-scale reactors. One soil was heavily contaminated with copper, the other with cadmium and arsenic (2500 microg g(-1) Cu; 300-400 microg g(-1) Cd and 230 microg g(-1) As, respectively). Test reactors with two separated chambers, each with a capacity of 5.25 kg soil, were constructed, then the respective chambers were filled with either a mixture of the polluted soil and a control topsoil (75:25) or topsoil alone. Reactors were sown with perennial ryegrass (Lolium perenne cv Elka) and a constant voltage of 30 V was applied continually across the soils in each reactor. Soil sampling took place at the start and the end of the test run, whilst plant foliage was sampled after approximately 3 weeks (both reactors) 6 weeks (Cd soil reactor only) and at the conclusion of each test run (98 days Cu soil, 80 days Cd soil). Soil and plant metal concentrations were measured, together with soil pH. Results showed that in both soils there was a significant re-distribution of metals from anode to cathode in the test reactors, coupled with an enhancement of plant Cu uptake in the cathode region for the Cu soil. Patterns of plant Cd uptake were less clear cut and were not as clearly related to the redistribution of Cd measured in the soil. There was significant acidification of soil at the anode in each test reactor, but soil pH in other parts of the reactor changed little during the course of the experiment. Plant growth was affected at the anode, but was not affected in other parts of the reactor. There was no visual evidence of metal toxicity in the ryegrass in either polluted soil. Some effects on soil fungi were apparent, with a stimulation of Fusarium infection of ryegrass in the cathode region of all reactors and the appearance of sporophores of Coprinus in the same location. It is concluded that the combination of the two techniques represents a very promising approach to the decontamination of metal polluted soils that now requires validation in field conditions. PMID- 12733818 TI - Pilot investigations of surface parts of three closed landfills and factors affecting them. AB - Aftercare of closed sanitary landfills in a major environmental problem. Rehabilitation of the landfill with vegetation and reducing leachate production are two issues that must be dealt. For this reason, Finnish Environment Institute has conducted several projects on closed landfills. This research aims at determining the physical and chemical properties of the soils at three closed landfills in Helsinki, Finland. Research was conducted to understand the impact by studying the following properties: Chemical, nutrient metal, gamma and radon analysis of surface soils of three closed landfills in Helsinki area. PMID- 12733819 TI - Mechanisms and consequences of large artery rigidity. AB - In this review paper, the classical and more recently described mechanisms responsible for the structural and functional characteristics of large artery rigidity are described. Mostly important, these characteristics appear to be non specific to the primary disease process involved in arterial hypertension, diabetes mellitus, dyslipidemia, congestive heart failure, chronic uremia, and perhaps senescence, including vascular dementia. Nonspecific in terms of aetiology, the vasculopathy encountered in these diseases exhibits common structural and functional abnormalities. The identification of such abnormalities could well become the target of potent nonpharmacological and (or) pharmacological interventions capable of preventing or retarding morbidity and mortality. The structural characteristics responsible for large artery rigidity include smooth muscle cell hypertrophy, matrix collagen deposition, and recently described, dysfunction in proteoglycan metabolism. Functional abnormalities, such as bradykinin-dependent hyper-reactivity of smooth muscle cells and vasa vasorum microcirculation network disturbances, also appear to alter aortic wall rigidity. The physiopathology of target organ damage is then revisited, based on endothelial dysfunction, documented in large and resistance arteries, as well as in microcirculation networks, where altered permeability to macromolecules leads to interstitial matrix disorganization and cell damage. The clinical evaluation of large artery rigidity is described, and one of the noninvasive methods, evaluation of pulse-wave velocity, is validated in normal conditions and in disease processes. Finally, non-pharmacological and pharmacological therapeutic measures are presented, and includes physical exercise to reduce insulin resistance, and renin-angiotensin-II-aldosterone modulators. PMID- 12733820 TI - Characterization of the NTPDase activities in the mesentery pre- and post capillary circuits of the guinea pig. AB - NTPDase is one of the principal enzymes involved in the sequential hydrolysis of ATP. In the present study, the presence and functionality of NTPDase in the mesenteric vein and artery were examined. Adenosine triphosphate (ATP) (0.01-1000 pmol) induces a dose-dependent vasodilation in the isolated arterial and venous mesenteric vasculatures of the guinea pig. Adenosine diphosphate (ADP) (0.01-1000 pmol) but not adenosine monophosphate (AMP) (0.01-1000 pmol) induces a similar response in the mesenteric vascular circuit. L-NAME, a nitric oxide synthase inhibitor (200 microM, 30 min), significantly reduces the arterial dilatory effect of ATP and abolishes the responses to ADP and AMP. Complete removal of the endothelium with 3-[(3-cholamidopropyl) dimethylammonio]-1-propansulfonate (CHAPS) (20 mM, 2 x 45 s) abolishes ATP-induced responses. Infusion of ATP in the vascular circuit generated detectable amounts of ADP and AMP, as measured by HPLC. CHAPS treatment significantly reduced the level of ATP and the production of AMP in the arterial mesenteric circuit. In contrast to the arterial mesenteric vasculature, endothelium removal in the venous circuit triggered a marked potentiation of ADP release and, interestingly, a marked reduction in the release of AMP. Moreover, a specific inhibitor of NTP diphosphohydrolase, 1 hydroxynaphthlene-3,6-disulfonic acid BGO 136 (10 mM for 20 min), significatively reduced AMP production in both vascular preparations. These results confirm that the endothelium contributes to the vasoactive properties of ATP, ADP, and AMP. Our data also demonstrated a significant role of endothelium in NTPDase activity on ADP and AMP production prior to exogenous administration of ATP. The activity of this particular enzyme appears to be different from the reaction products viewpoint (i.e., the production of ADP) in the pre- and post-mesenteric circuits, suggesting two different isoforms with different substrate specificities. PMID- 12733821 TI - RyR1/SERCA1 cross-talk regulation of calcium transport in heavy sarcoplasmic reticulum vesicles. AB - We investigated the functional interdependence of sarco-endoplasmic reticulum Ca2+ ATPase isoform 1 and ryanodine receptor isoform 1 in heavy sarcoplasmic reticulum membranes by synchronous fluorescence determination of extravesicular Ca2+ transients and catalytic activity. Under conditions of dynamic Ca2+ exchange ATPase catalytic activity was well coordinated to ryanodine receptor activation/inactivation states. Ryanodine-induced activation of Ca2+ release channel leaks also produced marked ATPase activation in the absence of measurable increases in bulk free extravesicular Ca2+. This suggested that Ca2+ pumps are highly sensitive to Ca2+ release channel leak status and potently buffer Ca2+ ions exiting cytoplasmic openings of ryanodine receptors. Conversely, ryanodine receptor activation was dependent on Ca2+-ATPase pump activity. Ryanodine receptor activation by cytosolic Ca2+ was (i) inversely proportional to luminal Ca2+ load and (ii) dependent upon the rate of presentation of cytosolic Ca2+. Progressive Ca2+ filling coincided with progressive loss of Ca2+ sequestration rates and at a threshold loading, ryanodine-induced Ca2+ release produced small transient reversals of catalytic activity. These data indicate that attainment of threshold luminal Ca2+ loads coordinates sensitization of Ca2+ release channels with autogenic inhibition of Ca2+ pumping. This suggests that Ca2+-dependent control of Ca2+ release in intact heavy sarcoplasmic reticulum membranes involves a Ca2+-mediated "cross-talk" between sarco-endoplasmic reticulum Ca2+ ATPase isoform 1 and ryanodine receptor isoform 1. PMID- 12733822 TI - Modulation of intracellular Ca2+ via L-type calcium channels in heart cells by the autoantibody directed against the second extracellular loop of the alpha1 adrenoceptors. AB - The effects of methoxamine, a selective alpha1-adrenergic receptor agonist, and the autoantibody directed against the second extracellular loop of alpha1 adrenoceptors were studied on intracellular free Ca2+ levels using confocal microscopy and ionic currents using the whole-cell patch clamp technique in single cells of 10-day-old embryonic chick and 20-week-old fetal human hearts. We observed that like methoxamine, the autoantibody directed against the second extracellular loop of alpha1-adrenoreceptors significantly increased the L-type calcium current (I(Ca(L))) but had no effect on the T-type calcium current (I(Ca(T))), the delayed outward potassium current, or the fast sodium current. This effect of the autoantibody was prevented by a prestimulation of the receptors with methoxamine and vice versa. Moreover, treating the cells with prazosin, a selective alpha1-adrenergic receptor antagonist blocked the methoxamine and the autoantibody-induced increase in I(Ca(L)), respectively. In absence of prazosin, both methoxamine and the autoantibody showed a substantial enhancement in the frequency of cell contraction and that of the concomitant cytosolic and nuclear free Ca2+ variations. The subsequent addition of nifedipine, a specific L-type Ca2+ channel blocker, reversed not only the methoxamine or the autoantibody-induced effect but also completely abolished cell contraction. These results demonstrated that functional alpha1-adrenoceptors exist in both 10-day-old embryonic chick and 20-week-old human fetal hearts and that the autoantibody directed against the second extracellular loop of this type of receptors plays an important role in stimulating their activity via activation of L-type calcium channels. This loop seems to have a functional significance by being the target of alpha1-receptor agonists like methoxamine. PMID- 12733824 TI - Angiotensin II and its receptors in human endocardial endothelial cells: role in modulating intracellular calcium. AB - The aims of the present study are to investigate the presence and distribution of angiotensin II (Ang II), as well as AT1 and AT2 receptors, in endocardial endothelial cells (EECs) and to determine if the effect of Ang II on intracellular calcium in these cells is mediated via the AT1 or the AT2 receptor. Immunofluorescence and 3D confocal microscopy techniques were used on 20-week-old fetal human EECs. Our results showed that Ang II and its receptors, the AT1 and the AT2 types, are present and exhibit a different distribution in human EECs. Ang II labelling is found throughout the cell with a fluorescence signal higher in the cytosol when compared with the nucleus. Like Ang II, the AT1 receptor fluorescence signal is also homogeneously distributed in human EECs but with a preferential labelling at the level of the nucleus, while the AT2 receptor labelling is solely present in the nucleus. Using fluo-3 and 3D confocal microscopy technique, superfusion of human EECs with increasing concentration of Ang II induced a dose-dependent sustained increase in free cytosolic and nuclear Ca2+ levels. This effect of Ang II on human EEC's intracellular Ca2+ ([Ca2+]) was completely prevented by losartan, an AT1 receptor antagonist. Our results suggest that Ang II, as well as AT1 and AT2 receptors, is present but differentially distributed in EECs of 20-week-old fetal human hearts, and that the AT1 receptor mediates the effects of Ang II on [Ca2+]i in these cells. PMID- 12733825 TI - Endothelium and myocyte cellular insulin receptor alterations in a rat model of myocardial infarction. AB - Ischemic heart disease is considered to be one of the leading causes of death in adults. While extensive research on mechanisms contributing to the pathogenesis of myocardial infarction (MI) has been underway, it is not known whether insulin receptor characteristics and postreceptor signaling have been fully addressed as yet. Present work attempts to investigate whether the remodeling process effectively induces alteration(s) in insulin-binding characteristics at the coronary endothelium and cardiomyocytes using a rat heart model of MI. MI was induced by ligation of the left anterior descending coronary artery of adult male Sprague-Dawley rats. Two animal groups were used in the study: (i) sham-operated CHAPS-untreated and CHAPS-treated, and (ii) MI CHAPS-untreated and MI CHAPS treated. A physical model describing 1:1 stoichiometry of reversible insulin binding to its receptors present on the endothelium and at cardiomyocytes after CHAPS treatment was considered for data analysis. Quantitation of the collected effluents after heart perfusion, the inlet at the aortic and outlet at the coronary sinus sites, were curve fitted using a first-order Bessel function, which determines the binding constants (k(n)), the reversible constant (k(-n)), the dissociation constant (k(d) = k(-n)/k(n)), and the residency time constant (tau = 1/k(-n)). In addition, hearts were excised, separated into right and left ventricles, and individually weighed, and areas of infarcted regions were measured. Results of the MI group showed significant increases in relative heart mass, left ventricle mass, and right ventricle mass normalized to total body mass. MI induced severe ischemia and irreversible myocardial injury as assessed by planimetry and histologic studies. The data showed differences in insulin receptor affinities at the endothelial and cardiac myocytes in the sham and in the MI-operated rats. The observed reduction in the binding affinity of insulin at the myocyte postinfarction may explain the pathogenic role of insulin in ischemic heart disease and, hence, resistance. Therefore, insulin administration during and post MI might be cardioprotective. PMID- 12733823 TI - Bradykinin induced a positive chronotropic effect via stimulation of T- and L type calcium currents in heart cells. AB - Using Fluo-3 calcium dye confocal microscopy and spontaneously contracting embryonic chick heart cells, bradykinin (10(-10) M) was found to induce positive chronotropic effects by increasing the frequency of the transient increase of cytosolic and nuclear free Ca2+. Pretreatment of the cells with either B1 or B2 receptor antagonists (R126 and R817, respectively) completely prevented bradykinin (BK) induced positive chronotropic effects on spontaneously contracting single heart cells. Using the whole-cell voltage clamp technique and ionic substitution to separate the different ionic current species, our results showed that BK (10(-6) M) had no effect on fast Na+ inward current and delayed outward potassium current. However, both L- and T-type Ca2+ currents were found to be increased by BK in a dose-dependent manner (10(-10)-10(-7) M). The effects of BK on T- and L-type Ca2+ currents were partially blocked by the B1 receptor antagonist [Leu8]des-Arg9-BK (R592) (10(-7) M) and completely reversed by the B2 receptor antagonist D-Arg[Hyp3,D-Phe7,Leu8]BK (R-588) (10(-7) M) or pretreatment with pertussis toxin (PTX). These results demonstrate that BK induced a positive chronotropic effect via stimulation of T- and L-type Ca2+ currents in heart cells mainly via stimulation of B2 receptor coupled to PTX-sensitive G-proteins. The increase of both types of Ca2+ current by BK in heart cells may explain the positive inotropic and chronotropic effects of this hormone. PMID- 12733827 TI - Presence of neuropeptide Y and the Y1 receptor in the plasma membrane and nuclear envelope of human endocardial endothelial cells: modulation of intracellular calcium. AB - The aims of the present study were to investigate the presence and distribution of NPY and the Y1 receptor in endocardial endothelial cells (EECs), to verify if EECs can release NPY, and to determine if the effect of NPY on intracellular calcium is mediated via the Y1 receptor. Immunofluorescence, 3-D confocal microscopy and radioimmunoassay techniques were used on 20-week-old human fetal EECs. Our results showed that NPY and the Y1 receptor are present in human EECs (hEECs) and that their distributions are similar, the fluorescence labelling being higher in the nucleus and more particularly at the level of the nuclear envelope when compared with the cytosol. Using radioimmunoassay, we demonstrated that EECs are a source of NPY and can secrete this peptide upon a sustained increase of intracellular calcium ([Ca]i). Using fluo-3 and 3-D confocal microscopy technique, superfusion of hEECs as well as EECs isolated from rat adult hearts with increasing concentrations of NPY induced a dose-dependent, sustained increase in free cytosolic and nuclear Ca2+ levels. This effect of NPY on EEC [Ca]i was completely reversible upon washout of NPY and was partially blocked by BIBP3226, a selective Y1 receptor antagonist. The results suggest that NPY and Y1 receptors are present in the EECs of 20-week-old human fetal heart and they share the same distribution and localization inside the cell. In addition, EECs are able to secrete NPY in response to an increase in [Ca]i, and the Y1 receptor as well as other NPY receptors seem to participate in mediating the effects of NPY on [Ca]i in these cells. Thus, NPY released by EECs may modulate excitation-secretion coupling of these cells. PMID- 12733826 TI - Angiotensin II AT1 receptor internalization, translocation and de novo synthesis modulate cytosolic and nuclear calcium in human vascular smooth muscle cells. AB - The present study was designed to verify if human (h) Angiotensin II (Ang II) type-1 receptor (hAT1R) undergoes internalization, nuclear translocation, and de novo synthesis in primary culture of human aortic vascular smooth muscle cells (hVSMCs) and if overexpression of this receptor modulates sustained free cytosolic ([Ca]c) and nuclear ([Ca]n) calcium. 3-dimensional (3-D) confocal microscopy was used to monitor free intracellular Ca2+ and hAT1R-green fluorescence protein (GFP) fusion protein in cultured hVSMCs. Immunofluorescence studies showed the presence of hAT1R and the absence of hAT2R in normal hVSMCs. Using 3-D imaging technique, hAT1 receptors were localized at the sarcolemma and in the cytosolic and nuclear compartments. In native as well as in normal hAT1R or hAT1R-GFP overexpressing hVSMCs, Ang II (10(-9) and 10(-4) M) induced internalization and nuclear translocation of this type of receptor. The internalization of hAT1Rs is mediated via clathrin-coated pits and vesicles pathway. This phenomenon of trancellular trafficking of receptors was associated with an increase of hAT1R. The Ang II induced increase of hAT1R density was prevented by the protein synthesis inhibitor cycloheximide. Overexpression of hAT1R and hAT1R-GFP decreased both basal cytosolic and nuclear Ca2+. In normal hVSMCs and low hAT1R-GFP overexpressing hVSMCs, Ang II (10(-15) to 10(-4) M) induced a dose-dependent sustained increase of [Ca]c and [Ca]n with an EC50 near 5 x 10(-11) and 5 x 10(-9) M, respectively. Our results suggest that hAT1Rs are the predominant type of Ang II receptors in aortic hVSMCs and are present in the sarcolemma, the cytosolic and the nuclear compartments. Ang II rapidly induces hAT1R internalization, nuclear translocation, as well as nuclear de novo synthesis of this receptor. The hAT1R overexpression in hVSMCs modulates sustained [Ca]c and [Ca]n. PMID- 12733828 TI - Nucleoplasmic calcium regulation in rabbit aortic vascular smooth muscle cells. AB - In this study, we investigated whether nucleoplasmic free Ca2+ in aortic vascular smooth muscle cells (VSMCs) might be independently regulated from cytosolic free Ca2+. Understanding mechanisms and pathways responsible for this regulation is especially relevant given the role of a numerous intranuclear Ca2+-sensitive proteins in transcriptional regulation, apoptosis and cell division. The question of an independent regulatory mechanism remains largely unsettled because the previous use of intensitometric fluorophores (e.g., Fluo-3) has been criticized on technical grounds. To circumvent the potential problem of fluorescence artifact, we utilized confocal laser scanning microscopy to image intracellular Ca2+ movements with the ratiometric fluorophore Indo-1. In cultured rabbit VSMCs, we found sarcoplasmic reticulum (SR) Ca2+ ATPase (SERCA) pumps and ryanodine receptor (RyR) Ca2+ channel proteins to be discretely arranged within a perinuclear locus, as determined by fluorescent staining patterns of BODIPY FL thapsigargin and BODIPY FL-X Ry. When intracellular Ca2+ stores were mobilized by addition of thapsigargin (5 microM) and activatory concentrations of ryanodine (1 microM), Indo-1 ratiometric signals were largely restricted to the nucleoplasm. Cytosolic signals, by comparison, were relatively small and even then its spatial distribution was largely perinuclear rather homogeneous. These observations indicate perinuclear RyR and SERCA proteins are intimately involved in regulating VSMC nucleoplasmic Ca2+ concentrations. We also observed a similar pattern of largely nucleoplasmic Ca2+ mobilization upon exposure of cells to the immunosuppressant drug FK506 (tacrolimus), which binds to the RyR-associated immunophillin-binding proteins FKBP12 and FKBP12.6. However, initial FK506 induced nucleoplasmic Ca2+ mobilization was followed by marked reduction of Indo 1 signal intensity close to pretreatment levels. This suggested FK506 exerts both activatory and inhibitory effects upon RyR channels. The latter was reinforced by observed effects of FK506 to only reduce nucleoplasmic Indo-1 signal intensity when added following pretreatment with both activatory and inhibitory concentrations of ryanodine. These latter observations raise the possibility that VSMC nuclei represent an important sink of intracellular Ca2+ and may help explain vasodilatory actions of FK506 observed by others. PMID- 12733829 TI - Do we really need real epidemiological scientific meetings? PMID- 12733830 TI - Trends in mortality sex ratios. PMID- 12733831 TI - Gender differentials in life expectancy in Italy. AB - OBJECTIVE: To analyze the gender differential in life expectancy in Italy from 1970 to 1997, to determine which ages and causes of death mostly influenced its trend. METHODS: Pollard method was used to explain which ages and which pathologies give the highest contribution to the gender differential. RESULTS: A slight reduction of the gender differential has been observed since 1980. The olds and very olds became the most relevant contributors to the differential. Cardiovascular diseases were the principal contributing causes of death until 1980 and Cancer replaced them afterwards. CONCLUSION: Relatively recent adoption of unhealthy life styles by women together with an opposite process run by young men towards healthier behaviour, seem to be at the basis of the slight reduction of the gender differential in life expectancy observed since 1980. PMID- 12733832 TI - Twenty-five-year coronary mortality trends in the seven countries study using the accelerated failure time model. AB - The 'accelerated failure time model' (AFT) was tested in the 25-year experience of the Seven Countries Study, to describe the shape of hazard for coronary heart disease (CHD) mortality during long-term follow-up. Sixteen cohorts of men aged 40-59 at entry were enrolled in eight countries (USA, Finland, The Netherlands, Italy, Croatia, Serbia, Greece and Japan) for a total of 12,763 subjects. Risk factors were measured and CHD mortality data collected during 25 years. The AFT incorporating the Weibull survival distribution was applied to the pooled cohorts of each country, with CHD mortality as endpoint. Cumulative hazards and the annual hazard for CHD mortality were estimated during 25 years and compared among countries. The annual hazard for CHD in 25 years tended to reduce relatively or flatten out in the northern American and northern European countries, exponentially increased in Serbia and Japan, and increased moderately in the other countries of southern Europe. Positive correlations were found between the shape of the hazard curve and population mean risk factor changes (serum cholesterol and systolic blood pressure) observed during the first 10 years follow-up. The Japanese group was an outlier. The countries with the largest relative decline in the annual hazard function were the same where, during the same historical period, the largest decreases in official death rates from CHD were recorded and vice versa. The curve shape in the annual hazard for CHD mortality in different countries seems related to changes in mean levels of serum cholesterol and systolic blood pressure. PMID- 12733833 TI - Risk of sporadic amyotrophic lateral sclerosis associated with seropositivity for herpesviruses and echovirus-7. AB - We examined the association between risk of sporadic amyotrophic lateral sclerosis (ALS) and seroprevalence of antibodies to echovirus-7 (echo-7) and herpesviruses 6, 7, and 8 through a population-based case-control study. We enrolled in a northern Italy area 20 newly diagnosed ALS cases and 20 referents. Risk of ALS was higher in subjects seropositive for echo-7 when we used the immunofluorescent assay, while little increase was noted with the neutralization test. Considering the different characteristics of these two serological assays, these results suggest an association between disease risk and infection with enterovirus (EV) family members (not specifically echo-7). ALS risk was slightly associated with seropositivity of human herpesvirus-6 (odds ratio: 3.2; p = 0.102) and more strongly with human herpesvirus-8 seropositivity (odds ratio: 8.4; p = 0.064), though these point estimates were statistically unstable due to the limited number of observed cases. The findings of this study warrant further investigation in larger studies of the possible etiologic role of EV or herpesvirus infection in sporadic ALS. PMID- 12733834 TI - Social networks and risk of neural tube defects. AB - The contribution of social support and social networks to risk of neural tube defects (NTDs) has not been explored, despite evidence that various aspects of the social environment contribute to their etiology. Using data from a population based case-control study of deliveries occurring in California from 1989 to 1991, this study investigates whether social networks, as measured by the presence and frequency of various types of social contacts, are associated with NTD risk. In person interviews were conducted with 538 (88% of eligible) mothers of NTD cases and with mothers of 539 non-malformed controls (88%) on average 5 months from the term delivery date. Unadjusted results indicate that women who reported having close friends, close relatives, or monthly contacts with close friends or relatives had 30-70% lower risks of having NTD-affected pregnancies, relative to women reporting 'none' to any of these measures. Only the presence of close friends was associated with reduced NTD risk after adjustment for other aspects of the social network and several potential covariates. This study is the first to investigate the association between social networks and risk of NTDs. Stronger inferences await replication and extension of these results. PMID- 12733835 TI - Ethnic distribution of Chlamydophila pneumoniae antibodies in a Malaysian population and possible correlation with coronary heart disease. AB - Chlamydophila pneumoniae, an important respiratory pathogen causing lower respiratory tract infections, has also been implicated in coronary heart disease (CHD). This study reports a cross-sectional, demographic, serological analysis of the prevalence of Chlamydophila pneumoniae antibodies in a multiracial Malaysian population. Generally, Malaysian Indians had the highest degree of seropositivity (58%) followed by the Chinese (54%) and the Malays (32%), results which were statistically significant (CI: 95%; p < 0.01). Interestingly, this trend was also reflected in the study group consisting of patients with acute myocardial infarctions (AMI) and chronic CHD. Again, the Indians were more frequently seropositive (65%), with more than 50% having IgG titres > or = 128. Comparatively, the Chinese and Malays showed 51 and 17% seropositivity respectively. These results were also statistically significant (CI: 95%; p < 0.01). Malaysian Indians are more commonly afflicted with CHD. A variety of factors have been suggested to explain this prevalence including diet, social habits, genetics and the possible role of infectious agents. This study notes an interesting association between this pattern of racial prevalence and the possible role of C. pneumoniae infections as a contributory/predisposing factor in the development of cardiovascular disease. PMID- 12733837 TI - Distribution and relationships between clinical and biopathological parameters in canine leishmaniasis. AB - Although the lesions, clinical signs and biochemical alterations observed in the course of canine leishmaniasis have been amply described a thorough definition and characterisation of the affected populations is important in order to detect relationships between parameters which may be involved in the development of this disease and to correctly assess further studies. This study included 61 dogs diagnosed with naturally acquired canine leishmaniasis by means of indirect immunofluorescence assay (IFA). At diagnosis, dogs were classified according to the following qualitative and quantitative variables: gender, breed, age, clinical picture, antibody titre, serum protein electrophoretogram, haemogram (CBC), urea, creatinine and ALT. Our population included dogs of 24 breeds, both sexes and different ages indicating no sex, age or breed predilection. In relation to the clinical picture, eight dogs were asymptomatic, 16 displayed mainly cutaneous signs, 18 presented primarily visceral signs and 19 displayed both cutaneous and visceral signs. Our results indicate that the clinical picture is significantly related to electrophoretogram and to RBC, PCV and haemoglobin. Dogs with mainly cutaneous signs showed the highest eosinophil mean values and those with mainly visceral signs showed the highest alpha-globulin mean values. This study confirms that the antibody titre is highly correlated with electrophoretogram and with RBC, PCV and haemoglobin. Lymphocytes were not associated or correlated with any other variable considered. PMNC, monocytes and eosinophils, as well as WBC, showed a significant correlation with beta globulins, which is difficult to interpret. PMID- 12733836 TI - Distribution of mecA among methicillin-resistant clinical staphylococcal strains isolated at hospitals in Naples, Italy. AB - Two hundred and twenty strains of Staphylococcus isolated in Naples, Italy, were surveyed for the distribution of the mecA, the structural gene for penicillin binding protein 2a, which is the genetic determinant for methicillin-resistance in staphylococci. Screening by a cloned mecA, revealed that of 220 strains, 43 were methicillin-resistant (19.5%) and 177 were methicillin-susceptible (80.5%). Among the 43 resistant strains 23 (53.5%) carried mecA in their genome and 20 (46.5%) did not carry mecA, in spite of their resistance to methicillin. Every group was submitted to the AP-PCR profiling. A quantitative analysis of the patterns divided strains into four different clusters for methicillin-resistant mecA-negative and two different clusters for methicillin-resistant mecA-positive with primer 1, while no clusters were noted with primer 7. We conclude that these clinical isolates from our area, were not found to belong to a single clone, although the predominance of four methicillin-resistant mecA-negative genotypes were noted. PMID- 12733838 TI - Verocytotoxin-producing Escherichia coli in foodstuffs of animal origin. AB - While much research effort has been targeted at the verocytotoxin-producing Escherichia coli (VTEC) serotype O157:H7, it is becoming more evident that other VTEC serotypes can also be associated with human foodborne disease. An increasing number of these non-O157 serotypes have been isolated from food sources and from humans suffering from haemolytic-uraemic syndrome and diarrhoea. The aim of our work was to investigate the prevalence of VTEC O157 and non-O157 in foodstuffs of animal origin using two rapid enzymatic procedures. Various types of food samples, 352 in total, were tested: 233 with the Premier EHEC, a screening test which directly detects the presence of verocytotoxin, regardless of serotype, while 119 of these with the Vidas ECO, which is a specific screening test for E. coli O157:H7, together with the Premier EHEC. Two samples were positive for VTEC, one of serogroup O126 and the other was non-serotypable. Another sample was positive in the test specific for E. coli O157:H7, but was not confirmed by culture. This study suggests that VTEC strains are not prevalent in Italy, and that the isolation of serogroup O157 is relatively infrequent. This leads us to conclude that there is little chance of exposure to pathogen for the average consumer in Italy. PMID- 12733840 TI - A one-generation cluster of malignant mesothelioma within a family reveals exposure to asbestos-contaminated jute bags in Naples, Italy. AB - Substantial evidence supports the role of asbestos in malignant mesothelioma. Clustering for this malignancy among relatives not only suggests genetic susceptibility as a relevant component but also provides a clue to investigate non-occupational sources of exposure. We identified five cases of malignant mesothelioma within one family with exposure to asbestos experienced during childhood, as 'next door' residents of a workshop recycling asbestos-contaminated jute sacks in Naples, Italy. This cluster discloses the health risk in the reuse of bags that previously had contained asbestos. Furthermore, it emphasizes the role of asbestos in the genetic-environmental interaction issue of malignant mesothelioma. PMID- 12733839 TI - Fully vaccinated children are rare: immunization coverage and seroprevalence in Austrian school children. AB - Vaccination coverage for vaccine-preventable diseases in Austria as well as in many Central European countries has been reported to be too low to eradicate such diseases and prevent further outbreaks. Austria lacks an adequate surveillance system to monitor prevalence of the diseases, the vaccination coverage and seroconversion. School children aged 10-14 years (n = 1077) were recruited in all four schools in the city of Schwaz, Austria, to present their vaccination documents and to give blood for serological testing (diphtheria, pertussis, measles, mumps, rubella, varicella). All participants received a report with a personal guideline for (re-) vaccination. Overall vaccination coverage was 86.4% for measles, 85.5% for mumps and 35.0% for rubella. Tetanus vaccination coverage was 98.4% for the first, 97.8% for the second and 96.7% for the third dose, while 55.4% of the study subjects received the recommended two booster injections. For diphtheria the corresponding vaccination coverage was found to be almost identical. Pertussis coverage was lower in general (first dose: 90.9%; second dose: 89.0%; third dose: 86.5%). Oral poliomyelitis vaccination showed a coverage of 98.6, 96.5, 95.3%, with 78.7% receiving the fourth dose. Overall 38.7% were classified as fully vaccinated. Seropositivity for measles was found in 90.4%, for mumps in 61.8%, for rubella in 82.3%, for diphtheria in 65.8%, for pertussis in 35.6% and for varicella in 95.0%. In summary, fully vaccinated children are rare and intensive public health efforts will be necessary to reach higher levels of immunity and prevent further outbreaks. PMID- 12733842 TI - Smoking expands expected lifetime with musculoskeletal disease. AB - By indirect estimation of mortality from smoking and life table methods we estimated expected lifetime without musculoskeletal diseases among never smokers, ex-smokers, and smokers. We found that although life expectancy of a heavy smoker is 7 years shorter than that of a never smoker, heavy smokers can expect to live more than 2 years longer with musculoskeletal diseases than never smokers. PMID- 12733841 TI - Low socio-economic status and familial occurrence of goitre are associated with a high prevalence of goitre. AB - The occurrence of goitre is dependent on genetic and environmental factors, but the associations with socio-economic and life-style factors have only been examined briefly. A cohort of 4649 participants from the general population was examined with questionnaires, thyroid ultrasonography, clinical examination and blood tests. Data were analysed in linear models and logistic regression analysis. Thyroid volume and serum thyroglobulin were closely associated with educational level with higher values in the group with the lowest levels of education (p < 0.001). The same pattern applied to thyroid multinodularity at ultrasonography (p = 0.002) and palpable goitre (p = 0.01). Physical activity in leisure time was negatively associated with thyroid enlargement (p = 0.02) and serum thyroglobulin (p < 0.001). These associations diminished markedly if adjustment was made for smoking habits, alcohol consumption and iodine intake. Familial occurrence of goitre was associated with goitre prevalence (Odds Ratio 2.5, 95% CI: 1.6-3.9), but did not confound the socio-economic associations. In conclusion, social imbalances in the occurrence of goitre were identified. These imbalances could in part be explained by differences in smoking habits and iodine intake. PMID- 12733843 TI - Of consents and CONSORTs: reporting ethics, law, and human rights in RCTs involving monitored overdose of healthy volunteers pre and post the "CONSORT" guidelines. AB - Randomized controlled trials (RCTs) of therapeutic interventions in acute drug overdose present a significant challenge for ethical, legal, and human rights protections of research subjects, particularly when healthy volunteers are involved. The CONSORT statement on the uniform reporting of clinical trials was published in 1996 with the overall aim of improving the reporting of RCTs, both individually and to facilitate their inclusion into systematic reviews. In CONSORT, reporting of ethical, legal, and human rights protections, including prior evaluation of the study by an ethics committee and provision of informed consent, was largely an implicit requirement. Those drafting CONSORT may have assumed such protections and the rights of study subjects were secured by existing doctor-patient relationships. Alternatively, CONSORT may have been viewed as likely to indirectly enhance such protections, as a flow-on effect of improved RCT design and reporting. We wished to examine whether such assumptions were justified by examining the reporting of RCTs of simulated overdose in healthy volunteers. We reviewed all reported RCTs involving activated charcoal in healthy human volunteersfor three years before the CONSORT statement (1989, 1990, and 1991) and three years afterwards (1999, 2000, 2001). Presence of documentation of inclusion and exclusion criteria, stopping rules, protocol deviations, information sheets, consent documentation, ethical approvals, conflicts of interest, understanding, refusal, inducements and coercion were recorded. We found a very poor level of reporting of some key ethical, legal, and human rights protections for healthy volunteers in toxicological RCTs. Reporting did not improve with the publication of CONSORT even in relation to requirements specifically included in the guidelines. PMID- 12733844 TI - Cardiotoxicity associated with intentional ziprasidone and bupropion overdose. AB - BACKGROUND: Ziprasidone and bupropion are medications prescribed for mood and behavior disorders. They have apparently safe cardiac safety profiles in both therapeutic and supratherapeutic doses, but recently the Federal Drug Administration has issued a caution regarding ziprasidone use in combination with other drugs that are known to prolong the QTc interval. CASE REPORT: A 17-year old male developed a widened QRS and a prolonged QTc interval following an overdose of ziprasidone and bupropion. He required hospital admission for aggressive cardiac monitoring and antidysrhythmic therapy, stabilizing to baseline by 80 hours postingestion. CONCLUSIONS: We present a case that underscores the potential cardiotoxicities of these medications. Ziprasidone and bupropion ingestion can be associated with cardiotoxicities that may require several days of aggressive cardiac monitoring and treatment. PMID- 12733845 TI - Quetiapine cross-reactivity among three tricyclic antidepressant immunoassays. AB - Quetiapine is an atypical antipsychotic agent with structural similarities to the tricyclic antidepressants (TCA). We report a case of quetiapine overdose that was initially clinically similar to that of a TCA overdose and caused a false positive TCA immunoassay. We then analyzed three common TCA immunoassays [Microgenics (formerly Diagnostic Reagents, Inc.) Tricyclics Serum Tox EIA Assay, Syva RapidTest d.a.u., and Biosite Triage Panel for Drugs of Abuse] with quetiapine in solution as well as urine from both an overdose patient and a therapeutic patient. There was significant variation of the cutoff of false positivity in all three immunoassays. Both the Syva and Microgenics immunoassays tested positive in both the overdose and therapeutic samples and were positive at urine levels of 100 microg/mL and 10 microg/mL, respectively. The Triage immunoassay was negative in solutions up to 1000 microg/mL and negative in both the therapeutic and overdose urine samples. Quetiapine may cause false-positive TCA immunoassay with both therapeutic use and in overdose. Significant variation exists between immunoassays to detect quetiapine as a false-positive test. PMID- 12733846 TI - Acute hepatitis induced by kava kava. AB - BACKGROUND: Herbal preparations are available widely and regarded generally by the public as harmless remedies for a variety of medical ailments. We report a case of acute hepatitis associated with the use of kava kava, derived from the root of the pepper plant, Piper methysticum. It is used in the United States as an antianxiety and sedative agent. CASE REPORT: A previously healthy 14-year-old female was admitted to the hospital with hepatic failure. Initial therapy, including plasmapheresis, was unsuccessful and she deteriorated. She ultimately required a liver transplant and now remains well. The liver biopsy showed hepatocellular necrosis consistent with chemical hepatitis. A work-up for alternative causes of liver failure was negative. The patient gave a history of taking a kava kava-containing product for four months. The use of kava kava and liver failure, is supported by kava kava use, a negative work-up for alternative causes of liver failure, and histological changes in the liver. CONCLUSIONS: Health care professionals need to be aware of the possibility of kava kava induced hepatotoxicity. The toxicity of these alternative remedies emphasizes the importance of surveillance programs and quality control in the manufacture of these products. Clinicians must remain aware of the toxic potential of herbal products and always inquire about their intake in cases of unexplained liver injury. PMID- 12733847 TI - Do poison centers diagnose organic dust toxic syndrome? PMID- 12733848 TI - Pressure-controlled ventilation attenuates lung microvascular injury in a rat model of activated charcoal aspiration. AB - BACKGROUND: Previous animal data suggest that aspiration of activated charcoal is associated with pulmonary microvascular injury that may be related to excessive ventilator-induced airway pressures. The purpose of this study was to test the hypothesis that ventilator-induced airway trauma contributes to the lung vascular injury observed following activated charcoal aspiration. METHODS: Capillary filtration coefficient (Kf,c), a sensitive measure of lung microvascular permeability, was determined isogravimetrically prior to and after intratracheal instillation of 0.4 ml/kg (12% weight/vol. solution, pH 7.4) activated charcoal oran equal volume of sterile water in isolated, perfused rat lungs in which ventilation was either pressure-controlled at 10cm H2O or volume-controlled at 5 ml/kg. RESULTS: There was significant lung injury in both activated charcoal groups regardless of ventilation method compared to control lungs or lungs administered sterile water (p < 0.05 ANOVA). However, injury to pressure controlled ventilated lungs was significantly less than lungs ventilated with traditional, volume-controlled ventilation. CONCLUSION: The results of this investigation demonstrate that pressure-controlled ventilation reduces the lung microvascular injury observed following aspiration of activated charcoal as compared to traditional volume-controlled ventilation methods. PMID- 12733850 TI - The abrupt cessation of therapeutically administered sodium oxybate (GHB) does not cause withdrawal symptoms. AB - Sodium oxybate (gamma-hydroxybutyrate; GHB) has demonstrated efficacy for the treatment of narcolepsy. However, there are reports of withdrawal following chronic abuse of illicit GHB which involve escalating both doses and dosing frequency. The present trial afforded an opportunity to test the hypothesis that chronic daily therapeutic dosing of sodium oxybate in narcoleptics does not cause withdrawal following abrupt cessation. Fifty-five narcoleptic patients, taking sodium oxybate (dose range 3-9 gm/night) for 7-44 months (mean 21 months), were randomized into a 2-week double-blind period: 29 patients received placebo and 26 continued to receive sodium oxybate. During this 2-week trial period, the following symptoms were reported in patients receiving placebo (N): anxiety (2), dizziness (1), insomnia (1) and somnolence (1). While these symptoms may represent possible symptoms of mild GHB withdrawal, they are also highly consistent with the returning symptoms of narcolepsy. We conclude there is minimal evidence of withdrawal symptoms following abrupt cessation of chronic sodium oxybate dosing in the therapeutic range. PMID- 12733849 TI - Prospective study of morbidity associated with snakebite envenomation. AB - BACKGROUND: The morbidity associated with snakebite envenomation has not been well documented. METHOD: Using a standardized questionnaire all patients with snakebite reported to a regional poison center during the year 2001 were followed after hospital discharge by telephone until resolution of symptoms. RESULTS: One hundred and twenty-eight snakebite cases were reported, of which 16 (12.5%) were lost to follow-up and 31 (24.2%) reported no progression of symptoms beyond puncture and were deemed "dry bites." Eighty-one (63.3%) patients were followed for the duration of symptoms. Age ranged from 1 to 86 years with a mean of 32 years. There were 64 males (79%). The snakes were identified as copperhead (n = 57), unidentified venomous (n = 17), timber rattlesnake (n = 6), and cottonmouth (n = 1). All patients were initially evaluated in a hospital emergency department of which 51 were admitted. Nine patients received antivenin. Of the 37 patients who had a job, 33 lost a mean of 14 days of work (SD +/- 18.1). Mean duration of edema was reported as 11.4 days (S.D +/- 12). Recurrent edema frequently occurred with limb activity. Pain was scored on a scale of 1 to 10, with a mean score of 4.8 (SD +/- 2.7). Mean duration of pain was reported as 7.8 days (SD +/- 6.4). Thirty patients required accommodation for ambulation including crutches (n = 11), limp (n = 11), and no shoes or loose shoes (n = 14). Of the 26 patients bitten on the hand or finger, duration of reduced function persisted for a mean of 14.3 days (SD +/- 10.4) and reduction of hand strength persisted for a mean of 22 days (SD +/- 25.5). Five patients had poorly healing wounds at the bite site which persisted from 14 to 77 days with a mean of 45 days (SD +/- 22.8) CONCLUSION: In this study snakebite resulted in significant duration and extent of morbidity in a majority of patients. PMID- 12733851 TI - Dimercaptosuccinic acid and Prussian Blue in the treatment of acute thallium poisoning in rats. AB - INTRODUCTION: Despite being banned as a pesticide, thallium still results in human and animal poisonings. Current recommended treatments include the use of the chemical Prussian Blue. Limitations in its availability may result in Prussian Blue not being obtainable in the thallium-poisoned patient. The chelator 2,3-Dimercaptosuccinic acid (DMSA) is currently FDA-approved for use in childhood lead poisoning and has been reported to be beneficial in treating other heavy metal poisonings. The objective of this study was to determine the efficacy of DMSA as a treatment for thallium poisoning by studying mortality and whole-brain concentrations in thallium poisoned rats. MATERIAL AND METHODS: Rats were gavaged with 30 mg/kg of thallium. After 24 hours they were randomized to DMSA (n = 20) 50 mg/kg twice daily for 5 days, Prussian Blue (n = 20) 50 mg/kg twice daily for 5 days, or control (n = 30). Animals were monitored twice daily for weight loss and mortality. Animals losing greater than 20% of their starting weight were euthanized and counted as a mortality. All surviving rats at 120 hours had their brains harvested and digested and underwent subsequent thallium analysis. RESULTS: The rate of survival in DMSA-treated animals compared to control was 45% vs. 21%, p = 0.07. Mean whole-brain thallium concentrations between DMSA and control rats were 3.4 vs. 3.0 microg/g, p = 0.06. Prussian Blue-treated rats had significantly improved survival (70% vs. 21%, p < 0.01) and lower whole-brain thallium concentrations (1.6 vs. 3.0 microg/g, p < 0.01 tissue) compared to controls. CONCLUSION: DMSA failed to reduce brain thallium concentrations in rats poisoned with thallium and had an indeterminate effect on mortality while Prussian Blue significantly reduces both brain thallium concentrations and mortality. PMID- 12733852 TI - Dancing with "death": p-methoxyamphetamine overdose and its acute management. AB - Para-methoxyamphetamine (PMA) is a substituted synthetic amphetamine used in the recreational drug scene. It is unusual because of the high incidence of significant morbidity and mortality in overdose. We report a case of PMA overdose in South Australia, and review our experience with the drug. We review the literature on PMA overdose and offer suggestions on the management of overdose with this dangerous drug. PMID- 12733854 TI - Dendrotoxin poisoning in a neurobiochemist. AB - BACKGROUND: Dendrotoxin is a highly potent blocker of KV1.1, KV1.2, and KV1.6 potassium channels that is derived from the venom of the green mamba (Dendroaspis angusticeps). It is commonly used to inhibit the function of whole nerve preparations in vitro. Despite the widespread use and potency of this compound, neurotoxicity in humans has not been described from refined toxin. We report a case of dendrotoxin toxicity from dermal exposure. CASE REPORT: A healthy 40-year old female neurobiochemist presented with complaints of progressive numbness of the left malar region and lateral orbit that progressed to include the medial orbit and tongue. One hour prior to presentation she used her bare hands to remove residual petroleum jelly from a dish that had previously contained 500 nanoliters of 500 nanomolar dendrotoxin. She recalled rubbing her left eye prior to the onset of symptoms. Before touching the dish, she had washed it with running water and then 70% ethanol while using latex gloves. Physical examination was remarkable only for weakness to superior gaze and some mild tongue fasciculations. Within 12 hours of exposure, she was completely asymptomatic. CONCLUSION: Dendrotoxin is a highly potent neurotoxin that can cause localized impairment of nerve function after mucous membrane exposure. PMID- 12733853 TI - What are the adverse effects of ethanol used as an antidote in the treatment of suspected methanol poisoning in children? AB - BACKGROUND: Ethanol used as an antidote is said to have various adverse effects, particularly in children. The rate of these adverse effects is not known. METHODS: Twenty-one-year retrospective chart review (1980-2000) from suspected methanol poisoning patients treated with ethanol in two large pediatric tertiary care centers. RESULTS: A total of 60 children (median age of 24 months) received ethanol for suspected methanol poisoning: 39 orally and 21 intravenously. Median initial methanol level was 4.16 mmol/L (13.3 mg/dL) (range 0 to 87.5 mmol/L or 0 to 280 mg/dL). Median duration of ethanol treatment was 16 hours (range 1.5 to 72 hours). None [0% (95% CI 0-5%)] of the 60 patients developed symptomatic hypoglycemia. Of the 50 patients that had a glucose level measured, none [(0% [95% CI 0-6%)] had a serum glucose concentration < 2.78 mmol/L (< 50 mg/dL). Eight patients [16% (95% CI 8-30%)] had at least one serum glucose concentration between 2.78-3.61 mmol/L (50-65 mg/dL), but none of those had symptoms compatible with hypoglycemia. A total of 42 patients [84% (95% CI 70-92%)] had all their serum glucose concentrations > 3.61mmol/L (> 65 mg/dL). There was no identifiable difference in the glucose intake between the serum glucose concentration groups. Six out of the 60 patients [10% (95% CI 4-21%)] were described as more drowsy after ethanol but none was comatose or needed intubation. No child showed signs of hypothermia [0/40 (95% CI 0-8%)] (rectal temperature < 35 degrees C), hepatotoxicity (0/12) (AST or ALT > 100 U/L) or even thrombophlebitis (0/21). None of the 22 patients with toxic levels of methanol (> or = 26.2 mmol/L- > or = 20 mg/dL) died or had ethanol-induced morbidity despite wide variation in ethanol levels. CONCLUSION: The rate of clinically important adverse effects related to ethanol used as an antidote to treat methanol poisoning in children was either absent or low in a tertiary care pediatric hospital setting. There was no morbidity or mortality associated with ethanol when it was used despite wide variation in ethanol levels. These results suggest that with appropriate monitoring and intravenous glucose intake in a controlled environment such as a pediatric intensive care unit, ethanol therapy does not carry as many risks as currently believed. PMID- 12733855 TI - Nefazodone poisoning: toxicokinetics and toxicodynamics using continuous data collection. AB - Nefazodone overdose has been reported infrequently. The commonest effects reported are drowsiness, nausea, dizziness, and vomiting, less commonly hypotension and bradycardia. We report a case of single-agent nefazodone poisoning. Serial drug concentrations were taken for quantification of parent drug and metabolites. Clinical findings were documented every 1 to 2 hours. We modeled both the toxicokinetics of nefazodone and correlated this with clinical effects and electrocardiograph (ECG) abnormalities. A 16-year-old female took 2.4 g of nefazodone. She had significant drowsiness in the first 6 hours, associated with hypotension (systolic BP < 90 mmHg; lowest BP 70/30 mmHg) for 18 hours, and mild bradycardia (slowest rate of 56 bpm). She had a prolonged QT/QTc which normalized over 24 hours. She was given charcoal and intravenous fluids and was observed carefully, recovering with no problems. The terminal elimination half life for nefazodone was 8.3 hours, and its metabolite hydroxy(OH)-nefazodone was 14.6 hours. BP-time curves demonstrated an 18-hour period of hypotension. There was a significant correlation between systolic BPand OH-nefazodone (R2 = 0.602). HR remained between 56 and 66 bpm for 30 hours despite hypotension. QT was significantly correlated with nefazodone (R2 = 0.911) and OH-nefazodone (R2 = 0.797), but no significant relationship between QTc and drug concentrations. The case demonstrates that nefazodone may potentially cause cardiac toxicity, which appears to be concentration-dependent. Although QT was concentration-dependent, this would need confirmation with other cases. Bradycardia, hypotension, and drowsiness are the most significant effects and are maximal in the first 12 hours when parent and metabolite concentrations are high. PMID- 12733856 TI - A patient with postoperative mercury contamination of the peritoneum. AB - CASE REPORT: Peritoneal exposure to mercury has been rarely reported and long term consequences of this type of exposure have not been documented. We report the clinical course of a patient who has survived almost eight years with a massive intraperitoneal load of mercury. She has suffered formication, pruritus, fatigue, irritiability, insomnia, alopecia, dizziness, a gait disturbance, loss of balance and multiple falls, abdominal pain, choking, and headaches. Two courses of chelation with dimercaptosuccinic acid using the standard protocol were undertaken, resulting in increased daily excretion, but without demonstrable objective or subjective benefit or lasting effect. She had multiple medical problems before the mercury intoxication, which complicates the attribution of all her problems to mercury intoxication. It is of particular interest that the patient survived and did not suffer any marked cognitive deterioration. She died in 2002 shortly after being diagnosed with lung cancer and declaring that she would fight it. Phasing out of mercury-weighted tubes is recommended. PMID- 12733857 TI - Bromism from daily over intake of bromide salt. AB - Bromide intoxication today is an infrequent disease, but preparations containing bromide are still available in nonprescription compounds, on the French market. We report a casewith bromide intoxication due to daily over intake (approximately 20 tablets per day; i.e. total elemental bromide intake approximately 6 g/day) of calcium bromo-galactogluconate (Calcibronat) for 1.5 months. A 30-year-old woman with a long history of psychotropic drug abuse was hospitalized in a psychiatric department for neuropsychological manifestations. She presented a seriously disturbed mental status with confusion, disorientation, auditory and visual hallucinations, and loss of short-time memory. A markedly increased serum bromide level of 1717 mg/L (21.5 mEq/L) measured on the first day after her admission confirmed the diagnosis of chronic bromism suspected based on her symptomatology. During her hospitalization, bromide plasma concentrations were measured and monitored using inductively coupled plasma mass spectrometry, a sensitive and very specific method. After withdrawal of the drug, the symptoms improved within 8 days. Serial bromide concentrations gradually declined throughout nearly 2 months of monitoring, until she was discharged from the hospital. We found an elimination half-life of bromide in blood of approximately of 10 days. This case demonstrates that, while today bromism occurs infrequently, it should still be included in the differential diagnosis of neuropsychiatric symptoms. PMID- 12733859 TI - Dietary aflatoxin exposure and chemoprevention of cancer: a clinical review. AB - Exposure to dietary aflatoxins is considered to be an important risk factor for the development of hepatocellular carcinoma in certain regions of the world. Significant advances have recently been made in understanding the clinical toxicology of aflatoxins. These include the development and validation of biomarkers of exposure and genotoxic effect. These biomarkers are currently being utilized to explore the potential that pharmaceutical interventions may have in modifying the toxicokinetics of dietary aflatoxin exposure. Preliminary results of clinical trials with the drug oltipraz suggest that it may modify the genotoxic effects of aflatoxin B1 by inhibiting bioactivation pathways and stimulating detoxification pathways. More recent results of a clinical trial with chlorophyllin suggest that this drug may have a role in preventing dietary exposure to aflatoxin B1 by reducing its oral bioavailability. The preliminary results of these chemoprevention studies may ultimately have implications for cancer prevention in high-risk populations in the future. PMID- 12733858 TI - Organic dust toxic syndrome: a review. AB - Because of differences in presentation, clinical course, diagnostic testing, treatments, prognosis, followup requirements and prevention strategies between ODTS, hypersensitivity pneumonitis, oxides of nitrogen and other pulmonary exposure-related illnesses, it is important for poison centers and clinicians to obtain appropriate elements of history, physical examination, and laboratory data that will allow an accurate diagnosis. PMID- 12733860 TI - Occupational exposure to cephalosporins leading to clostridium difficile infection. PMID- 12733861 TI - Unintentional confusion of semantics is not accidental. PMID- 12733863 TI - Advancing HIV prevention: new strategies for a changing epidemic--United States, 2003. AB - In several U.S. cities, recent outbreaks of primary and secondary syphilis among men who have sex with men (MSM) and increases in newly diagnosed human immunodeficiency virus (HIV) infections among MSM and among heterosexuals have created concern that HIV incidence might be increasing. In addition, declines in HIV morbidity and mortality during the late 1990s attributable to combination antiretroviral therapy appear to have ended. Until now, CDC has mainly targeted its prevention efforts at persons at risk for becoming infected with HIV by providing funding to state and local health departments and nongovernmental community-based organizations (CBOs) for programs aimed at reducing sexual and drug-using risk behavior. Some recent programs have focused on prevention efforts for persons living with HIV. Funding HIV-prevention programs for communities heavily affected by HIV has promoted community support for prevention activities. At the same time, these communities recognize the need for new strategies for combating the epidemic. In addition, the recent approval of a simple rapid HIV test in the United States creates an opportunity to overcome some of the traditional barriers to early diagnosis and treatment of infected persons. Therefore, CDC, in partnership with other U.S. Department of Health and Services agencies and other government agencies and nongovernment agencies will launch a new initiative in 2003, Advancing HIV Prevention: New Strategies for a Changing Epidemic. PMID- 12733862 TI - Neuropsychologic impairment, MRI abnormalities, and solvent abuse. PMID- 12733864 TI - Update: Severe acute respiratory syndrome--United States, 2003. AB - CDC and the World Health Organization (WHO) are continuing to investigate the multicountry outbreak of severe acute respiratory syndrome (SARS). This report updates information on reported SARS cases among U.S. residents and summarizes information on patients with no recent travel outside the United States. PMID- 12733865 TI - Pneumoconiosis prevalence among working coal miners examined in federal chest radiograph surveillance programs--United States, 1996-2002. AB - Coal workers' pneumoconiosis (CWP) is a chronic lung disease caused by inhalation of coal mine dust. To characterize the prevalence of CWP, the National Institute for Occupational Safety and Health (NIOSH) analyzed recent radiographic information from the U.S. National Coal Workers' X-ray Surveillance Program (CWXSP). Established under the Federal Coal Mine Health and Safety Act of 1969, CWXSP is administered by NIOSH under federal regulations. NIOSH is responsible for approving coal miner examination plans, submitted approximately every 5 years by companies that operate underground coal mines. This report summarizes the results of the analysis, which indicate that the overall prevalence of CWP among participating miners continues to decline; however, new cases are occurring among miners who have worked exclusively under current dust exposure limits. An evaluation of the mining conditions that have resulted in these cases is underway. PMID- 12733866 TI - Preliminary FoodNet data on the incidence of foodborne illnesses--selected sites, United States, 2002. AB - In the United States, an estimated 76 million persons contract foodborne illnesses each year. CDC's Emerging Infections Program Foodborne Diseases Active Surveillance Network (FoodNet) collects data on 10 foodborne diseases in nine U.S. sites. FoodNet follows trends in foodborne infections by using laboratory based surveillance for culture-confirmed illness caused by several enteric pathogens commonly transmitted through food. This report describes preliminary surveillance data for 2002 and compares them with 1996-2001 data. The data indicate a sustained decrease in major bacterial foodborne illnesses such as Campylobacter and Listeria, indicating progress toward meeting the national health objectives of reducing the incidence of foodborne infections by 2010 (objectives 10-1a to 10-1d). However, the data do not indicate a sustained decline in other major foodborne infections such as Escherichia coli O157 and Salmonella, indicating that increased efforts are needed to reduce further the incidence of foodborne illnesses. PMID- 12733867 TI - Update: Adverse events following civilian smallpox vaccination--United States, 2003. AB - During January 24-April 13, 2003, smallpox vaccine was administered to 32,644 civilian health-care and public health workers in 54 jurisdictions as part of an effort to prepare the United States for a possible terrorist attack using smallpox virus. This report updates information on all vaccine-associated adverse events among civilians vaccinated since the beginning of the vaccination program and among contacts of vaccinees, received by CDC from the Vaccine Adverse Event Reporting System (VAERS) as of April 13. PMID- 12733868 TI - The biosynthesis of GDP-L-colitose: C-3 deoxygenation is catalyzed by a unique coenzyme B6-dependent enzyme. AB - l-Colitose (1) is a 3,6-dideoxyhexose found in the O-antigen of gram-negative lipopoly-saccharides. While the biosynthesis of many deoxysugars have previously been investigated, l-colitose is distinct in that it originates from GDP-d mannose. In contrast, other 3,6-dideoxyhexoses arise from CDP-d-glucose. Therefore, the enzymes involved in the l-colitose biosynthetic pathway must be specifically tailored to utilize such a modified substrate. The mode for deoxygenation at C-3 of colitose is of particular interest because this conversion in other naturally occurring 3,6-dideoxyhexoses requires a pair of enzymes, E1 and E3, acting in concert. Interestingly, no E3 equivalent was identified in the five open reading frames of the col biosynthetic gene cluster from Yersinia pseudotuberculosis IVA. However, the gene product of colD showed moderate similarity with the E1 gene (ddhC/ascC) of the ascarylose pathway (27% identity and 42% similarity). Because E1 is a pyridoxamine 5'-phosphate (PMP) dependent enzyme, it was thought that ColD might also utilize PMP. Indeed, turnover was observed during incubation of ColD with substrate in the presence of excess PMP, but not with pyridoxal 5'-phosphate (PLP). However, the rate of product formation increased by more than 40-fold when l-glutamate was included in the PLP incubation. The formation of alpha-ketoglutarate as a byproduct under these conditions clearly indicated that ColD functions as a transaminase, recognizing both PMP and PLP. In this paper, we propose a novel biosynthetic route for colitose, including the unprecedented C-3 deoxygenation performed solely by ColD. The utilization of PMP in a dehydration reaction is rare, but the combined deoxygenation-transamination activity makes ColD a unique enzyme. PMID- 12733869 TI - Structural biasing elements for in-cell histone deacetylase paralog selectivity. AB - We use the structural dissection of two 1,3-dioxanes with in-cell histone deacetylase (HDAC) paralog selectivity to identify key elements for selective HDAC inhibitors. We demonstrate that o-aminoanilides are inactive toward HDAC6 while apparently inhibiting deacetylases that act upon histone substrates. This finding has important clinical implications for the development of HDAC inhibitor based treatments that do not interfere with microtubule dynamics associated with HDAC6. We also show that suberoylanilide hydroxamic acid (SAHA) alone is a nonparalog-selective HDAC inhibitor and that the 1,3-dioxane diversity appended to SAHA is essential for HDAC6 paralog selectivity. PMID- 12733871 TI - A remarkable [2.2.2]propellane. AB - To explore the effects of fluorine substitution on the highly strained [2.2.2]propellane skeleton, a new representative of this ring system, perfluorotricyclo[2.2.2.01,4]octan-2-one ethylene ketal, was prepared by a rapid and quantitative [2+2] cycloaddition to the strained alkene perfluorobicyclo[2.2.0]hex-1(4)-ene. The propellane displays impressive thermal stability, and the vulnerable C-C bond joining the bridgeheads is very resistant to attack by electrophilic reagents. On the other hand, that electron-deficient bond is cleaved quickly at room temperature by a variety of nucleophiles and mild reducing agents. The behavior of this compound contrasts dramatically with that of the only known [2.2.2]propellane lacking fluorine substituents. PMID- 12733870 TI - Protein nanostructures formed via direct-write dip-pen nanolithography. AB - We report a tip modification strategy coupled with dip-pen nanolithography for depositing proteins on a surface. This methodology offers patterning capabilities on the scale of 45 nm to many micrometers length. The biorecognition properties of nanofeatures composed of immunoglobulin-gamma (IgG) were confirmed by reacting the array with gold nanoparticles coated with anti-IgG. Furthermore, a two component nanoarray was fabricated in direct-write fashion by DPN, and the biorecognition properties of this structure were investigated by AFM without biodiagnostic labels. PMID- 12733872 TI - Environment-independent 14-helix formation in short beta-peptides: striking a balance between shape control and functional diversity. AB - We report a significant and unanticipated advance in the study of beta-amino acid based foldamers: a small proportion of highly preorganized residues can impart high stability to a specific helical secondary structure in water. Most of the residues in these beta-peptides (2 and 3) are intrinsically flexible. Flexible beta-amino acids can be readily and enantiospecifically prepared in functionally diverse forms, but preorganized residues with side chains are rare and challenging to synthesize. Our findings demonstrate that interspersing a few copies of an unfunctionalized but rigid residue among a larger number of flexible residues with diverse side chains is a viable strategy for creating beta-peptides that adopt the 14-helix conformation and therefore display side chains in a predictable spatial arrangement. These results are significant because they enhance the prospects of developing beta-peptides with useful activities. PMID- 12733873 TI - G-quadruplex-specific peptide-hemicyanine ligands by partial combinatorial selection. AB - G-quadruplex DNA-specific ligands were generated using a combinatorial selection of tetrapeptides which were then conjugated to a hemicyanine scaffold. The heterocycle enhanced peptide binding affinity by approximately 1000-fold to give ligands with near micromolar affinity and >40-fold discrimination for quadruplex DNA over duplex. PMID- 12733874 TI - Self-consistency analysis of dipolar couplings in multiple alignments of ubiquitin. AB - A self-consistency analysis of backbone N-H residual dipolar couplings of ubiquitin collected in 10 different media is described to assess the degree of structural and dynamic heterogeneous behavior across the media. The SECONDA method, which works with and without any structural or dynamic information about the molecular system, is based on a principal component analysis and is very sensitive to the presence of heterogeneities or experimental errors. It is found that the regular secondary structural elements behave highly homogeneously, while small heterogeneities are manifested in the loop region 51-63. Many residues that exhibit increased dynamics in NMR relaxation experiments are inert with respect to changes in the alignment. PMID- 12733875 TI - Charge optimization increases the potency and selectivity of a chorismate mutase inhibitor. AB - The highest affinity inhibitor for chorismate mutases, a conformationally constrained oxabicyclic dicarboxylate transition state analogue, was modified as suggested by computational charge optimization methods. As predicted, replacement of the C10 carboxylate in this molecule with a nitro group yields an even more potent inhibitor of a chorismate mutase from Bacillus subtilis (BsCM), but the magnitude of the improvement (roughly 3-fold, corresponding to a DeltaDeltaG of 0.7 kcal/mol) is substantially lower than the gain of 2-3 kcal/mol binding free energy anticipated for the reduced desolvation penalty upon binding. Experiments with a truncated version of the enzyme show that the flexible C terminus, which was only partially resolved in the crystal structure and hence omitted from the calculations, provides favorable interactions with the C10 group that partially compensate for its desolvation. Although truncation diminishes the affinity of the enzyme for both inhibitors, the nitro derivative binds 1.7 kcal/mol more tightly than the dicarboxylate, in reasonable agreement with the calculations. Significantly, substitution of the C10 carboxylate with a nitro group also enhances the selectivity of inhibition of BsCM relative to a chorismate mutase from Escherichia coli (EcCM), which has a completely different fold and binding pocket, by 10-fold. These results experimentally verify the utility of charge optimization methods for improving interactions between proteins and low molecular weight ligands. PMID- 12733876 TI - Liquid poly(ethylene glycol) and supercritical carbon dioxide: a benign biphasic solvent system for use and recycling of homogeneous catalysts. AB - Poly(ethylene glycol) (PEG), having a molecular weight of 900 or 1500, is a solid at room temperature but a nonvolatile liquid at 40 degrees C under CO2 pressure. Homogeneously catalyzed hydrogenation can be performed in the molten PEG, followed by extraction of the product by supercritical CO2. The catalyst containing PEG phase which remains in the vessel can be reused for hydrogenation without addition of further catalyst or PEG. PMID- 12733877 TI - Discovery of a new phosphotyrosine mimetic for PTP1B using breakaway tethering. AB - Protein tyrosine phosphatases play important roles in many signaling cascades involved in human disease. The identification of druglike inhibitors for these targets is a major challenge, and the discovery of suitable phosphotyrosine (pY) mimetics remains one of the key difficulties. Here we describe an extension of tethering technology, "breakaway tethering", which is ideally suited for discovering such new chemical entities. The approach involves first irreversibly modifying a protein with an extender that contains both a masked thiol and a known pY mimetic. The extender is then cleaved to release the pY mimetic, unmasking the thiol. The resulting protein is screened against a library of disulfide-containing small molecule fragments; any molecules with inherent affinity for the pY binding site will preferentially form disulfides with the extender, allowing for their identification by mass spectrometry. The ability to start from a known substrate mimimizes perturbation of protein structure and increases the opportunity to probe the active site using tethering. We applied this approach to the anti-diabetic protein PTP1B to discover a pY mimetic which belongs to a new molecular class and which binds in a novel fashion. PMID- 12733879 TI - Novel alkenylative cyclization using a ruthenium catalyst. AB - Novel alkenylative cyclization of enyne was developed using Cp*RuCl(cod) under ethylene gas at room temperature. PMID- 12733878 TI - The interstitial atom of the nitrogenase FeMo-cofactor: ENDOR and ESEEM show it is not an exchangeable nitrogen. AB - A recent high-resolution X-ray crystallographic study (1.16 A) of the Azotobacter vinelandii nitrogenase MoFe protein revealed a previously undetected electron density associated with the active site FeMo-cofactor. The density is located inside the cluster at the center of the "trigonal prism" of six irons and is assigned to a species "X". The identity of species X was not resolved, although the electron density is consistent with a single N, O, or C atom. One proposal is that X is an N atom that derives from and exchanges with N from N2 during catalysis. In the present study, we have examined this possibility by employing 14N and 15N isotopes of N2 along with ENDOR and ESEEM spectroscopies. The WT MoFe protein and alpha-359Arg-->Lys and alpha-381Phe-->Leu variants were allowed to turn over in the presence of 14N2 or 15N2, and then were examined as resting enzymes by ENDOR and ESEEM at X- and Q-bands to look for all 14N and 15N signals coupled to the electron spin of the FeMo-cofactor and to determine if any exchanged during turnover. We have found five peaks in Q-band pulsed ENDOR spectra that appear to arise not only from previously reported N1/N2, which give rise to the ESEEM, but also from one or two additional coupled nitrogens. None of the ENDOR and ESEEM signals vanish or are altered by catalytic turnover with 15N2, and no new 15N signal is detected, leading to the conclusion that if species X is a nitrogen atom, it does not exchange during dinitrogen reduction. PMID- 12733880 TI - Rhodium-catalyzed anti-Markovnikov hydroamination of vinylarenes. AB - The transition metal-catalyzed anti-Markovnikov hydroamination of unactivated vinylarenes with a rhodium complex of DPEphos is reported. The reaction of electron-neutral or electron-rich vinylarenes with a variety of secondary amines in the presence of catalyst forms the products from anti-Markovnikov hydroamination in high yields. Reactions of morpholine, N-phenylpiperazine, N-Boc piperazine, piperidine, 2,5-dimethylmorpholine, and perhydroisoquinoline reacted with styrene to form the amine product in 51-71% yield. Reactions of a variety of vinylarenes with morpholine generated amine as the major product. Reactions of morpholine with electron-poor vinylarenes gave lower amine:enamine ratios than reactions of electron-rich vinylarenes at the same concentration of vinylarene, but conditions were developed with lower concentrations of electron-poor vinylarene to maintain formation of the amine as the major product. Reactions of dimethylamine with vinylarenes were fast and formed amine as the major product. Mechanistic studies on the hydroamination process showed that the amine:enamine ratio was lower for reactions conducted with higher concentrations of vinylarene and that one vinylarene influences the selectivity for reaction of another. A mechanism proceeding through a metallacyclic intermediate that opens in the presence of a second vinylarene accounts for these and other mechanistic observations. PMID- 12733881 TI - Reversible light-driven redox switching of multifunctional dipolar ruthenium(III/II) pentaammine(4,4'-bipyridinium) complexes. AB - We report the first example of a molecular switch of multifunctional dipolar ruthenium(III/II) pentaammine-N-methyl-(4,4'-bipyridinium) complexes, exclusively driven by light. This is achieved by using a two-phase (water/benzene) system in which RuIII/II complexes are soluble only in the water phase. The reversible redox switching is triggered by the selective irradiation of the water and the benzene compartments with 254 and 528 nm light, respectively. PMID- 12733882 TI - Light-driven open-close motion of chiral molecular scissors. AB - The first example of "light-driven chiral molecular scissors" (1), which consists of 1,1',3,3'-tetraarylferrocene as a pivot part and azobenzene as a driving part, was synthesized. Absorption, circular dichroism (CD), and 1H NMR spectral studies on the photoinduced isomerization process of an enantiomer of 1 agreed well with a prediction by a DFT calculation, where a motion of the handles via light-driven contraction/expansion of the connecting azobenzene strap was transformed, through a pivotal motion of the ferrocene unit, into an open-close motion of the blade parts. PMID- 12733883 TI - Generation and characterization of highly strained dibenzotetrakisdehydro[12]annulene. AB - We report here the generation of tetrakisdehydro[12]annulene possessing a highly deformed triyne component from the [4.3.2]propellatriene-annelated precursor by its photolysis extruding indan and the characterization of the highly reactive annulene by chemical and spectroscopic methods. In addition to the chemical evidence for the formation of the title compound in solution such as interception as a Diels-Alder adduct, we succeeded in its characterization by UV-vis and FTIR spectra in an argon matrix at 20 K. The experimental IR spectrum agreed well with the theoretical one calculated by the DFT method. PMID- 12733884 TI - Room-temperature Hiyama cross-couplings of arylsilanes with alkyl bromides and iodides. AB - The first method for achieving Hiyama couplings of unactivated alkyl bromides and iodides is reported. The desired carbon-carbon bond formation proceeds under mild conditions (room temperature) with good functional-group tolerance. PMID- 12733885 TI - A catalytically active, charge-neutral Rh(I) zwitterion featuring a P,N substituted "naked" indenide ligand. AB - The synthesis and characterization of a new class of cationic and zwitterionic Rh(I) complexes, which feature multidentate ligands comprised of donor functionalized indene or indenide units, are reported. This unusual new ligation strategy provides access to the first charge-neutral [kappa2-P,N]Rh(I) zwitterion, a complex that functions as a catalyst for the C-H/Si-H dehydrogenative coupling of styrene and triethylsilane. PMID- 12733886 TI - New insights into ethene epoxidation on two oxidized Ag[111] surfaces. AB - Reaction mechanisms and activation energies for the complete conversion of ethene to ethene epoxide on two recently characterized oxidized Ag{111} surfaces have been determined from density functional theory. On both surfaces, epoxidation proceeds through a two-step nonconcerted mechanism via an oxametallacycle intermediate. The key implications are that both surfaces are active and that epoxidation can take place over a wide O coverage regime. PMID- 12733887 TI - Cationic organoscandium beta-diketiminato chemistry: arene exchange kinetics in solvent separated ion pairs. AB - Abstraction of methide from the beta-diketiminato supported organoscandium complex [L1ScMe2]2 using the trityl borate activator [Ph3C][B(C6F5)4] in arene solvents gives solvent separated ion pairs in which the arene (C6H5Br, 1a; C6H6, 1b; C7H8, 1c; 1,3,5-Me3C6H3, 1d) is coordinated to the cationic scandium center in an eta6 bonding mode. L1 incorporates methyl groups in the 2,4 positions of the ligand backbone and bulky 2,6-diisopropylphenyl groups on the nitrogen atoms. The relative binding strength of the arenes is C6H5Br < C6H6 < 1,3,5-Me3C6H3 < C7H8. Ion pairs 1a and 1c have been characterized crystallographically, and the C6H5Br derivative is notable for its eta6 bonding mode in preference to the more common eta1 bonding mode via the halogen atom. The kinetics of displacement of mesitylene by toluene (1d --> 1c) yield activation parameters of DeltaH = 21.4(6) kcal mol-1 and DeltaS = 6(1) cal mol-1 K-1. In combination with the observed lack of dependence of [toluene] on the rate of displacement, these data suggest a mechanism involving partial dissociation of the coordinated arene, followed by attack of the incoming arene. This chemistry has relevance to the role of these solvent separated ion pairs in olefin polymerization processes and presents a rare opportunity for the detailed study of these ephemeral species. PMID- 12733888 TI - Multielectron transfer in clusters of polar-polarizable chromophores. AB - We present a model for interacting polar-polarizable molecules. Exotic collective effects are recognized as being due to supramolecular interactions: in attractive lattices, bistable behavior is found, and, for proper molecular orientation, multielectron transfer represents the primary photoexcitation event. PMID- 12733889 TI - Self-assembly of an environmentally responsive polymer/silica nanocomposite. AB - Thermoresponsive nanocomposite thin films composed of alternating layers of silica and polymerized N-isopropylacrylamide (NIPAM) or NIPAM plus dodecyl methacrylate (DM) hydrogels were prepared by surfactant-directed evaporation induced self-assembly (EISA). During EISA, the organic monomers partition within the hydrophobic domains of a lamellar mesophase. In-situ polymerization via a free radical process results in a 1-2 nm thick hydrogel phase sandwiched between layers of silica oriented parallel to the substrate surface. The thermoresponsiveness of PNIPAM is preserved in this confined environment, and the polymeric layers reversibly swell and deswell by a factor of 2 in water upon temperature changes around the transition temperature of PNIPAM (32 degrees C). The composition, mesostructure, and environmental response were studied by detailed NMR, TGA, and SAXS analyses. PMID- 12733890 TI - Short, enantioselective total synthesis of okaramine N. AB - The first enantioselective total synthesis of a member of the okaramine family of bis-indole alkaloids, okaramine N (1), has been accomplished via intermediates 2 7, as outlined. The N-prenylated derivative of (S)-tryptophan methyl ester (2) was coupled with Fmoc-protected N-tert-prenylated tryptophan (3) to form the amide 4 in 70% yield. Pd(II)-mediated cyclization/rearrangement, a key step in the synthesis, transformed 4 into the indoloazacine 5 (44%), which was deprotected and cyclized in a single step to give the hexacyclic diketopiperazine 6 (95%). In the following novel and key sequence, 6 was transformed into 1: (1) selective ene reaction with N-methyltriazolinedione, (2) photooxidation of the remaining tert-prenylated indole subunit to provide 7, and (3) thermal retroene reaction of 7 to afford okaramine N (70% from 6). PMID- 12733891 TI - The first oxovanadium ring in [[O[double bond]V(salen)]2(mu F)][VO(salen)][BF4].(CH2Cl2)x crystals. AB - The crystal structure of [{O=VV(salen)}2(mu-F)][VIVO(salen)][BF4].(CH2Cl2)x revealed a hollow cavity with a diameter of 5.3 A that penetrates through the crystal, and a remarkable cyclic chain of the [VO(salen)] unit, a motif that has never been fashioned from oxometal building blocks. These features endow the crystal with a molecular sievelike property for the rapid, reversible, and size selective absorption of guest CH2Cl2 molecules. PMID- 12733892 TI - Broad-spectrum radical cyclizations boosted by polarity matching. Carbonylative access to alpha-stannylmethylene lactams from azaenynes and CO. AB - Free-radical mediated stannylcarbonylation of azaenynes provides a general [n + 1]-type annulation approach leading to alpha-stannylmethylene lactams. The cyclization is unusual in its breadth, covering 4-exo, 5-exo, 6-exo, 7-exo, and 8 exo modes. PMID- 12733894 TI - Growth of millimeter-long and horizontally aligned single-walled carbon nanotubes on flat substrates. AB - Millimeter-long and well-aligned single-walled carbon nanotubes (SWNTs) have been produced on silica/silicon surfaces using the carbon monoxide chemical vapor deposition (CO-CVD) method. The orientation of the nanotube arrays can be well controlled by the gas flow during the growth. The majority of the orientated SWNTs are straight and individual. The length of the nanotubes can be >2 mm for a 10 min growth. Furthermore, multidimensional crossed-networks of SWNT can be easily generated by multistep processes. These results present a great opportunity in the controllable production of organized SWNT arrays for large scale carbon nanotube-based nanodevice fabrication. PMID- 12733895 TI - Solventless synthesis of copper sulfide nanorods by thermolysis of a single source thiolate-derived precursor. AB - Organic monolayer protected Cu2S nanorods, 4 nm in diameter and 12 nm long, were synthesized using a novel solventless synthetic approach. Thermolytic degradation of a copper thiolate precursor at temperatures ranging from 140 to 200 degrees C produces Cu2S nanorods. Higher temperatures promote isotropic growth of spherical nanocrystals. X-ray diffraction and high-resolution TEM reveal that the nanorods exhibit a hexagonal Cu2S crystal structure, which in the bulk is ferroelectric. The appropriate reaction conditions produce nanorods that are size and shape monodisperse and organize into smectic superlattices. The extent of superlattice ordering and the appearance of extended strands of nanorods provide evidence for strong dipole-dipole coupling between Cu2S nanorods. PMID- 12733893 TI - Catalytic enantioselective Strecker reaction of ketoimines. AB - A new method for the catalytic enantioselective Strecker reaction (cyanation) of N-diphenylphosphinoyl ketoimines is described. The asymmetric catalyst is a chiral gadolinium complex prepared from Gd(OiPr)3 and the d-glucose-derived ligand 3 in a 1:2 ratio. The reaction has a broad substrate generality, giving high enantioselectivity from aromatic, ethyl, primary alkyl, and alpha,beta unsaturated ketoimines. The products could be easily converted to disubstituted alpha-amino acids and their derivatives. PMID- 12733896 TI - Photoaquation of methylated cis-dichlorobis(1,10 phenanthroline)rhodium(III)chloride compounds by direct population of a photoactive triplet excited state. AB - Photoaquation in compounds II and III by direct excitation into a photoactive triplet excited state is reported. The location of the singlet to triplet transition in compound III is estimated by a combination of the action spectrum for photoaquation in the region between 520 and 600 nm and the phosphorescence spectrum at 77 K. The significant increment of the reactivity (10-fold) of the triplet states in II and III as compared to that in I is explained in terms of increasing sigma-donation from the phen ligands stabilizing the pentacoordinated rhodium intermediate formed by chloride expulsion. PMID- 12733897 TI - Chromium-mediated asymmetric synthesis of both enantiomers of acetoxytubipofuran. AB - Both enantiomers of acetoxytubipofuran were synthesized using enantioselective and diastereoselective dearomatization sequences starting from the benzaldehyde chromium tricarbonyl complex. Following aldol condensation, a sequence involving Pd-catalyzed allylic substitution was used in the synthesis of the (-) enantiomer, whereas the (+)-enantiomer was reached via an Eschenmoser-Claisen rearrangement. Chiroptical data show that a revision of the previously assigned absolute configuration of the natural product is required. PMID- 12733898 TI - A new method for the catalytic aldol reaction to ketones. AB - A new method for the catalytic aldol reaction to ketones, using CuF.3PPh3.2EtOH complex as the catalyst and (EtO)3SiF as the additive, is described. The reaction can be applied to a wide range of ketones and trimethylsilyl enolates. On the basis of mechanistic studies, a working hypothesis for the catalytic cycle is proposed, in which the dynamic ligand exchange mediated by copper silicates produces the active copper enolate. Moreover, the present reaction can be extended to the catalytic enantioselective reaction using tol-BINAP as a chiral ligand. PMID- 12733899 TI - Ni- or Cu-catalyzed cross-coupling reaction of alkyl fluorides with Grignard reagents. AB - n-Octyl fluoride underwent a cross-coupling reaction with n-propylmagnesium bromide in the presence of 1,3-butadiene using NiCl2 as a catalyst at room temperature to give undecane in moderate yields. This alkyl-alkyl cross-coupling proceeded more efficiently when CuCl2 was employed instead of NiCl2. Addition of 1,3-butadiene dramatically improved the yields of the coupling products from primary alkyl Grignard reagents in both Ni- and Cu-catalyzed reactions. Alkyl fluorides efficiently reacted with tertiary alkyl and phenyl Grignard reagents using CuCl2 in the absence of 1,3-butadiene to afford the coupling products in high yields. The competitive reaction of a mixture of alkyl halides (R-X; X = F, Cl, Br) with nC5H11MgBr showed that the reactivities of the halides increase in the order R-Cl < R-F < R-Br. In contrast, in the Cu-catalyzed reaction with PhMgBr, the reactivities increase in the order R-Cl < R-Br < R-F. PMID- 12733900 TI - Measurements of carbon to amide-proton distances by C-H dipolar recoupling with 15N NMR detection. AB - A new magic-angle spinning NMR method for measuring internuclear distances between a 13C-labeled site and amide protons is described. The magnetization of the protons evolves under homonuclear decoupling and the recoupled 13C-1H dipolar interaction, which provides simple spin-pair REDOR curves if only one 13C-labeled site is present. The modulation of the amide proton HN is detected via short 1H 15N cross polarization followed by 15N detection. The method is demonstrated on two specifically 13C- and 15N-labeled peptides, with 13C-HN distances from 2.2 to ca. 6 A. This technique promises to be particularly useful for measuring distances between 13C=O and H-15N groups, to identify hydrogen bonds in peptides and proteins. PMID- 12733902 TI - Synthesis of tin-encapsulated spherical hollow carbon for anode material in lithium secondary batteries. AB - The tributylphenyltin (TBPT)-encapsulated resorcinol (R)-formaldehyde (F) sol was prepared inside the micelles of cetyltrimethylammonium bromide (CTAB). This core shell-type sol was polymerized and further carbonized to obtain nanosized Sn encapsulated spherical hollow carbon. The size of spherical hollow carbon and Sn metal particles was controllable by changing the R/CTAB or TBPT/CTAB mole ratio, respectively. It is likely that, when tested as the anode in Li secondary batteries, the spherical hollow carbon acts as a barrier to prevent the aggregation of nanosized Sn particles and provides a void space for Sn metal particles to experience a volume change without a collapse of carbon shell, giving rise to a better cycle performance than that of pure Sn metal. PMID- 12733901 TI - Micelle-encapsulated carbon nanotubes: a route to nanotube composites. AB - We report a general approach toward dispersing single-walled carbon nanotubes (SWNTs) in solvents and polymer materials, by encapsulating SWNTs within cross linked micelles. Micelles made from polystyrene-block-poly(acrylic acid) (PS-b PAA), an amphiphilic block copolymer, are first assembled around SWNTs by gradually adding H2O to a suspension of nanotubes in dimethylformamide. The hydrophilic, outer shells of these micelles are then chemically cross-linked with a difunctional linker molecule. Pure encapsulated SWNTs (e-SWNTs) can then be separated from empty cross-linked micelles by consecutive cycles of centrifugation and redispersion. Atomic force and transmission electron microscopies of the resulting nanostructures demonstrate that individual nanotubes (rather than bundles) have been completely encased in polymer shells whose thickness is slightly larger than that of empty micelles. e-SWNTs encapsulated in PS-b-PAA can be permanently redispersed in H2O, in organic solvents, and in both hydrophobic and hydrophilic polymer matrices with minimal sonication. Micelle encapsulation could improve the compositing of SWNTs in a wide variety of polymer materials for structural, electronic, and thermal applications. PMID- 12733903 TI - Four-stranded DNA structure stabilized by a novel G:C:A:T tetrad. AB - The solution structure of a cyclic oligonucleotide d has been determined by two-dimensional NMR spectroscopy and restrained molecular dynamics. Under the appropriate experimental conditions, this molecule self-associates, forming a symmetric dimer stabilized by four intermolecular Watson-Crick base pairs. The resulting four-stranded structure consists of two G:C:A:T tetrads, formed by facing the minor groove side of the Watson-Crick base-pairs. Most probably, the association of the base-pairs is stabilized by coordinating a Na(+) cation. This is the first time that this novel G:C:A:T tetrad has been found in an oligonucleotide structure. This observation increases considerably the number of sequences that may adopt a four-stranded architecture. Overall, the three dimensional structure is similar to those observed previously in other quadruplexes formed by minor groove alignment of Watson-Crick base pairs. This resemblance strongly suggests that we may be observing a general motif for DNA DNA recognition. PMID- 12733904 TI - Selection of a high-energy bioactive conformation of a sulfonium-ion glycosidase inhibitor by the enzyme glucoamylase G2. AB - Transferred nuclear Overhauser effect and rotating-frame Overhauser enhancement NMR spectroscopies are used to probe the conformation of a bicyclic sulfonium ion, which is an analogue of the naturally occurring glycosidase inhibitor castanospermine, bound to the enzyme glucoamylase G2. Enzyme inhibition assays indicate that the bicyclic sulfonium ion is a slightly better inhibitor (K(i) = 1.32 mM) of glucoamylase G2 than the naturally occurring sulfonium-ion glycosidase inhibitor, salacinol, with a K(i) value of 1.7 mM. The NMR results are interpreted in terms of the selection by the enzyme of a high-energy conformation of the ligand that is already represented in the ensemble of free ligand conformations. PMID- 12733905 TI - Expression and kinetic analysis of the substrate specificity of modules 5 and 6 of the picromycin/methymycin polyketide synthase. AB - Picromycin synthase (PICS) is a multifunctional, modular polyketide synthase (PKS) that catalyzes the conversion of methylmalonyl-CoA to narbonolide and 10 deoxymethynolide, the macrolide aglycone precursors of the antibiotics picromycin and methymycin, respectively. PICS modules 5 and 6 were each expressed in Escherichia coli with a thioesterase domain at the C-terminus to allow release of polyketide products. The substrate specificity of PICS modules 5+TE and 6+TE was investigated using N-acetylcysteamine thioesters of 2-methyl-3-hydroxy-pentanoic acid as diketide analogues of the natural polyketide chain elongation substrates. PICS module 5+TE could catalyze the chain elongation of only the syn diketide (2S,3R)-4, while PICS module 6+TE processed both syn diastereomers, (2S,3R)-4 and (2R,3S)-5, with a 2.5:1 preference in k(cat)/K(m) for 5 but did not turn over either of the two anti diketides. The observed substrate specificity patterns are in contrast to the 15-100:1 preference for 4 over 5 previously established for several modules of the closely related erythromycin PKS, 6-deoxyerythronolide B synthase (DEBS). PMID- 12733906 TI - Spectroscopic and kinetic studies of PKU-inducing mutants of phenylalanine hydroxylase: Arg158Gln and Glu280Lys. AB - Phenylalanine hydroxylase (PAH) is a tetrahydrobiopterin-dependent, nonheme iron enzyme that catalyzes the hydroxylation of L-Phe to L-Tyr in the rate-limiting step of phenylalanine catabolism. This reaction is tightly coupled in the wild type enzyme to oxidation of the tetrahydropterin cofactor. Dysfunction of PAH activity in humans leads to the disease phenylketonuria (PKU). We have investigated two PKU-inducing mutants, Arg158Gln and Glu280Lys, using kinetic methods, magnetic circular dichrosim (MCD) spectroscopy, and X-ray absorption spectroscopy (XAS). Analysis of the products produced by the mutant enzymes shows that although both oxidize pterin at more than twice the rate of wild-type enzyme, these reactions are only approximately 20% coupled to production of L Tyr. Previous MCD and XAS studies had demonstrated that the resting Fe(II) site is six-coordinate in the wild-type enzyme and converts to a five-coordinate site when both L-Phe and reduced pterin are present in the active site. Although the Arg158Gln mutant forms the five-coordinate site when both cosubstrates are bound, the Fe(II) site of the Glu280Lys mutant remains six-coordinate. These results provide insight into the PAH reaction and disease mechanism at a molecular level, indicating that the first step of the mechanism is formation of a peroxy-pterin species, which subsequently reacts with the Fe(II) site if the pterin is properly oriented for formation of an Fe-OO-pterin bridge and an open coordination position is available on the Fe(II). PMID- 12733907 TI - Site-specific synthesis and reactivity of oligonucleotides containing stereochemically defined 1,N2-deoxyguanosine adducts of the lipid peroxidation product trans-4-hydroxynonenal. AB - trans-4-Hydroxynonenal (HNE) is a major peroxidation product of omega-6 polyunsaturated fatty acids. The reaction of HNE with DNA gives four diastereomeric 1,N(2)-gamma-hydroxypropano adducts of deoxyguanosine; background levels of these adducts have been detected in animal tissue. Stereospecific syntheses of these four adducts at the nucleoside level have been accomplished. In addition, a versatile strategy for their site-specific incorporation into oligonucleotides has been developed. These adducts are destabilizing as measured by melting temperature when compared to an unadducted strand. The thermal destablization of the adducted 12-mers ranged from 5 to 16 degrees C and is dependent on the absolute stereochemistry of the adduct. The HNE adducts were also examined for their ability to form interstrand DNA-DNA cross-links when incorporated into a CpG sequence. We find that only one of the HNE stereoisomers formed interstrand DNA-DNA cross-links. PMID- 12733908 TI - Side chain assignments of Ile delta 1 methyl groups in high molecular weight proteins: an application to a 46 ns tumbling molecule. AB - A sensitive 3D NMR pulse scheme, (H)C(CA)NH-COSY, is presented for the assignment of (13)C(delta)(1) Ile chemical shifts in large perdeuterated, methyl-protonated proteins. The nonlinearity of branched amino acids, such as Ile, significantly degrades the quality of TOCSY schemes which transfer magnetization from methyl carbons to the backbone (13)C(alpha) positions, and in applications to high molecular weight proteins (correlation times on the order of 40-50 ns), this compromises the sensitivity of spectra used for methyl assignment. The experiment presented utilizes COSY-based transfer steps and refocuses undesirable (13)C (13)C scalar couplings that degrade the efficiency of TOCSY transfers. The (H)C(CA)NH-COSY scheme is tested on an (15)N,(13)C,(2)H-[Leu, Val, Ile (delta 1 only)]-methyl-protonated maltose binding protein (MBP)/beta-cyclodextrin complex at 5 degrees C (molecular tumbling time 46 +/- 2 ns), facilitating the assignment of (13)C(delta 1) chemical shifts for 18 of the 19 Ile residues for which backbone assignments were previously obtained. Both sensitivity and resolution of the resulting spectra are shown to be significantly better than those for a similar TOCSY-based approach. PMID- 12733909 TI - Imidazopyridine/Pyrrole and hydroxybenzimidazole/pyrrole pairs for DNA minor groove recognition. AB - The DNA binding properties of fused heterocycles imidazo[4,5-b]pyridine (Ip) and hydroxybenzimidazole (Hz) paired with pyrrole (Py) in eight-ring hairpin polyamides are reported. The recognition profile of Ip/Py and Hz/Py pairs were compared to the five-membered ring pairs Im/Py and Hp/Py on a DNA restriction fragment at four 6-base pair recognition sites which vary at a single position 5' TGTNTA-3', where N = G, C, T, A. The Ip/Py pair distinguishes G.C from C.G, T.A, and A.T, and the Hz/Py pair distinguishes T.A from A.T, G.C, and C.G, affording a new set of heterocycle pairs to target the four Watson-Crick base pairs in the minor groove of DNA. PMID- 12733910 TI - Quantitative formation of [2]catenanes using copper(I) and palladium(II) as templating and assembling centers: the entwining route and the threading approach. AB - Transition metal-mediated templating and self-assembly have shown powerful potentials for the synthesis of interlocked molecules. These two strategies were combined in designing and preparing a new type of coordination catenanes incorporating Cu(I) and Pd(II) metal centers. The ligand designed here contains a phenanthroline core and pyridine sidearms (compound 1). Using this phenanthroline pyridine conjugated ligand, two approaches were examined, which were shown to be surprisingly efficient for the catenane synthesis: the entwining route (entwining of two ligands around Cu(I) followed by Pd(II) clipping) and the threading approach (Cu(I)-templated threading of a cyclic ligand on an acyclic ligand followed by the Pd(II) clipping of the second ring). In the former method, stepwise treatment of 1 with Cu(CH(3)CN)(4)PF(6) (templating center) and enPd(NO(3))(2) (assembling center) gives rise to the quantitative formation of CuPd(2) catenane 18. In the latter method, Cu(I) templates the threading of phenanthroline-containing macrocycle 2 on ligand 1, which is followed by Pd(II) clipping to give hetero catenane 20. In both approaches, the formation of catenanes is convincing thanks to the strong templating effect of Cu(I), while the ring closure steps are efficiently furnished by Pd(II)-directed self assembly. PMID- 12733911 TI - Silylated cyclohexadienes as new radical chain reducing reagents: preparative and mechanistic aspects. AB - Various silylated 1,4-cyclohexadienes are presented as superior tin hydride substitutes for the conduction of various radical chain reductions. Debrominations, deiodinations, and deselenations can be performed using these environmentally benign reagents. Furthermore, Barton-McCombie-type deoxygenations using silylated cyclohexadienes are described. Radical cyclizations, ring expansions, and Giese-type addition reactions with the new tin hydride substitutes are presented. The polymerization of styrene can be regulated using silylated cyclohexadienes. Rate constants for hydrogen atom abstraction from two 1-silyl-cyclohexadienes by primary C-radicals were determined. The effects of the cyclohexadiene substituents on the reaction outcomes are discussed. Finally, qualitative EPR experiments on silyl radical expulsion from silylated cyclohexadienyl radicals are presented. PMID- 12733913 TI - Stereochemical memory versus Curtin-Hammett behavior in the rearrangement of 1,3 cyclopentanediyl radical cations derived from housanes through structural effects on conformational control. AB - The electron-transfer-catalyzed rearrangement of the housanes 1 affords regioselectively the two cyclopentenes 2 and 3 by 1,2-migration of a group at the methano bridge. Appropriate ring annelation in the intermediary cyclopentane-1,3 diyl radical cation 1(*+) changes the stereochemical course of the rearrangement from complete stereoselectivity (stereochemical memory) for the structurally simple housane 1b to partial loss of stereoselectivity through competing conformational interconversion for the tricyclic housane 1c. Additional cyclohexane annelation, as in the tetracyclic housane 1a, results in complete loss of stereocontrol through Curtin-Hammett behavior, as substantiated by the viscosity dependence on the product ratio of the rearrangement. Whereas in the radical cations 1b(*+) and 1c(*+) the 1,2-shifts (k(2) and k(3)) are faster than the conformational anti <==> syn change (k(1), k(-1)), the reverse applies for the radical cation 1a(*+). Such structural manipulation of conformational effects in radical cation rearrangements has hitherto not been documented. PMID- 12733912 TI - Highly efficient and selective epoxidation of alkenes by photochemical oxygenation sensitized by a ruthenium(II) porphyrin with water as both electron and oxygen donor. AB - Visible light irradiation of a reaction mixture of carbonyl-coordinated tetra(2,4,6-trimethyl)phenylporphyrinatoruthenium(II) (Ru(II)TMP(CO)) as a photosensitizer, hexachloroplatinate(IV) as an electron acceptor, and an alkene in alkaline aqueous acetonitrile induces selective epoxidation of the alkene with high quantum yield (Phi = 0.6, selectivity = 94.4% for cyclohexene and Phi = 0.4, selectivity = 99.7% for norbornene) under degassed conditions. The oxygen atom of the epoxide was confirmed to come from a water molecule by an experiment with H(2)(18)O. cis-Stilbene was converted into its epoxide, cis-stilbeneoxide, without forming trans-stilbeneoxide. trans-Stilbene, however, did not exhibit any reactivity. Under neutral conditions, an efficient buildup of the cation radical of Ru(II)TMP(CO) was observed at the early stage of the photoreaction, while an addition of hydroxide ion caused a rapid reaction with the cation radical to promote the reaction with reversion to the starting Ru(II)TMP(CO). A possible involvement of a higher oxidized state of Ru such as Ru(IV), Ru(V), Ru(VI) through a dismutation of the Ru(III) species was excluded by an experiment with Ru(VI)TMP(O)(2). Decarbonylation of the Ru complex was also proven to be invalid. A reaction mechanism involving an electron transfer from the excited triplet state of Ru(II)TMP(CO) to hexachloroplatinate(IV) and subsequent formation of OH( )-coordinated Ru(III) species, leading to an oxo-ruthenium complex as the key intermediate of the photochemical epoxidation, was postulated. PMID- 12733915 TI - The intramolecular, stereoselective addition of sulfoximine carbanions to alpha,beta-unsaturated esters. AB - ortho-Bromocinnamates can be coupled with methyl phenylsulfoximine to afford N arylsulfoximines in excellent yield. Treatment of these products with an amide base results in a completely stereoselective cyclization to afford enantiomerically pure benzothiazines. This reaction is stereospecific. PMID- 12733916 TI - Intramolecular reactions of alkynes with furans and electron rich arenes catalyzed by PtCl2: the role of platinum carbenes as intermediates. AB - 5-(2-Furyl)-1-alkynes react, with PtCl(2) as catalyst, to give phenols. On the basis of DFT calculations, a cyclopropyl platinacarbene complex was found as the key intermediate in the process. The cyclopropane and dihydrofuran rings of this intermediate open to form a carbonyl compound, which reacts with the platinum carbene to form an oxepin, which is in equilibrium with an arene oxide. When the reaction is carried out in the presence of water, dicarbonyl compounds are obtained, which support the proposed mechanism. Other cyclizations of alkynes with furans or electron-rich arenes give products of apparent Friedel-Crafts-type reactions, although these processes could also proceed by pathways involving the formation of cyclopropyl platinum carbenes. PMID- 12733914 TI - Mithramycin SK, a novel antitumor drug with improved therapeutic index, mithramycin SA, and demycarosyl-mithramycin SK: three new products generated in the mithramycin producer Streptomyces argillaceus through combinatorial biosynthesis. AB - To gain initial structure-activity relationships regarding the highly functionalized pentyl side chain attached at C-3 of mithramycin (MTM), we focused on a post-polyketide synthase (post-PKS) tailoring step of the MTM biosynthesis by Streptomyces argillaceus ATCC 12956, which was proposed to be catalyzed by ketoreductase (KR) MtmW. In this last step of the MTM biosynthesis, a keto group of the pentyl side chain is reduced to a secondary alcohol, and we anticipated the generation of an MTM derivative with an additional keto group in the 3-side chain. Insertional inactivation of mtmW, a gene located ca. 8 kb downstream of the mithramycin-PKS genes, yielded an S. argillaceus mutant, which accumulated three new mithramycin analogues, namely mithramycin SA, demycarosyl-mithramycin SK, and mithramycin SK (MTM-SK). The structures of these three compounds confirmed indirectly the proposed role of MtmW in MTM biosynthesis. However, the new mithramycin derivatives bear unexpectedly shorter 3-side chains (ethyl or butyl) than MTM, presumably caused by nonenzymatic rearrangement or cleavage reactions of the initially formed pentyl side chain with a reactive beta dicarbonyl functional group. The major product, MTM-SK, was tested in vitro against a variety of human cancer cell lines, as well as in an in vitro toxicity assay, and showed an improved therapeutic index, in comparison to the parent drug, MTM. PMID- 12733917 TI - A disilapentalene and a stable diradical from the reaction of a dilithiosilole with a dichlorocyclopropene. AB - The reaction of 1,1-dilithio-2,3,4,5-tetraphenylsilole (1) with 1,1-dichloro-2,3 diphenylcyclopropene (2) leads to the novel 1,4-disila-1,4-dihydropentalene (4), as well as an exceptionally stable diradical for which the structure 3 is suggested. The diradical is unreactive toward water, methanol, and chloroform; upon heating it transforms into 4. Structure 3 for the paramagnetic species is proposed on the basis of EPR data and theoretical calculations. The trans-trans isomer of diradical 3 was calculated to be more stable than its cis-cis isomer. The strong and stable EPR signal in the reaction mixture is probably due to the trans-trans isomer of diradical 3 in the triplet state. A reaction scheme describing the formation of 3 and 4 is presented. PMID- 12733918 TI - Bis(calix[4]diquinone) receptors: cesium- and rubidium-selective redox-active ionophores. AB - A new class of redox-active ionophore comprised of two calix[4]diquinone moieties connected through either alkylene or pyridylene linkages has been developed. Spectroscopic and electrochemical investigations, X-ray crystal structure analyses, and molecular modeling studies show butylene- and propylene-linked members of this family of redox-active receptors exhibit remarkable selectivity preferences and substantial electrochemical recognition effects toward cesium and rubidium cations. PMID- 12733919 TI - Synthesis, characterization, and coordination chemistry of the 2-azaphenalenyl radical. AB - The 2-azaphenalenyl radical 2 has been synthesized and characterized by ESR spectroscopy. Variable-temperature ESR measurements were carried out on both the phenalenyl (1) and the 2-azaphenalenyl (2) radicals. The phenalenyl radical 1 has the known propensity to dimerize at temperatures below 20 degrees C, but unexpectedly less so than originally reported. The first experimental measurement of bond dissociation enthalpy for the dimerization of the phenalenyl radical 1 was obtained in CCl(4) (11.34 +/- 0.11 kcal/mol) and toluene (9.8 +/- 0.7 kcal/mol). The 2-azaphenalenyl radical 2 does not show a propensity to dimerize over the measurable temperature range (220-330 K), but does so in the presence of Cu(hfac)(2) (hfac = hexafluoroacetylacetonate). The latter complex was characterized by X-ray crystallography. PMID- 12733920 TI - Benzoxazine oligomers: evidence for a helical structure from solid-state NMR spectroscopy and DFT-based dynamics and chemical shift calculations. AB - A combination of molecular modeling, DFT calculations, and advanced solid-state NMR experiments is used to elucidate the supramolecular structure of a series of benzoxazine oligomers. Intramolecular hydrogen bonds are characterized and identified as the driving forces for ring-shape and helical conformations of trimeric and tetrameric units. In fast MAS (1)H NMR spectra, the resonances of the protons forming the hydrogen bonds can be assigned and used for validating and refining the structure by means of DFT-based geometry optimizations and (1)H chemical-shift calculations. Also supporting these proposed structures are homonuclear (1)H[bond](1)H double-quantum NMR spectra, which identify the local proton-proton proximities in each material. Additionally, quantitative (15)N[bond](1)H distance measurements obtained by analysis of dipolar spinning sideband patterns confirm the optimized geometry of the tetramer. These results clearly support the predicted helical geometry of the benzoxazine polymer. This geometry, in which the N...H...O and O...H...O hydrogen bonds are protected on the inside of the helix, can account for many of the exemplary chemical properties of the polybenzoxazine materials. The combination of advanced experimental solid-state NMR spectroscopy with computational geometry optimizations, total energy, and NMR spectra calculations is a powerful tool for structural analysis. Its results provide significantly more confidence than the individual measurements or calculations alone, in particular, because the microscopic structure of many disordered systems cannot be elucidated by means of conventional methods due to lack of long-range order. PMID- 12733922 TI - Self-activated supramolecular reactions: effects of host-guest recognition on the kinetics of the Diels-Alder reaction of open-chain oligoether quinones with cyclopentadiene. AB - Diels-Alder reactions of acyclic oligoether-substituted quinones 1b, 1c, 2b, and 2c with cyclopentadiene were accelerated by the addition of alkali and alkaline earth metal perchlorates, and scandium trifluoromethane sulfonate (k(c)/k(f) = 1.2-23 for univalent cations, 11-1160 for divalent cations, and 1700-192 000 for Sc(3+), where k(c) and k(f) are the rate constants for the metal complexed and uncomplexed quinones, respectively). The shorter-armed 1a, 2a, and 3, however, exhibited no such acceleration effects. The rate accelerations can be rationalized by the FMO consequence in which the bound guest cation withdraws electron density from the quinone dienophile and lowers the LUMO energy suitable for the orbital interaction with the HOMO of cyclopentadiene. Despite the poor cation selectivity, these acyclic oligoether quinones showed larger rate accelerations than the relevant quinocrown ethers 4 (k(c)/k(f) = 1.3-3.0 for univalent cations, 5.0-160 for divalent cations, and 100-2020 for Sc(3+)). The effective electron withdrawal, which leads to the enhanced rate acceleration, can be caused by the direct interaction between the metal cation accommodated in the pseudo-cyclic oligoether linkage and the quinone carbonyl oxygen, as indicated by (1)H NMR spectroscopy. In addition, the larger rate enhancement is rather achieved in the complex with low binding constant K, because the strong encapsulation of metal cation by the oligoether chain diminishes the crucial interaction to the quinone carbonyl oxygen. As a whole, the smaller and higher valent cations tend to bring about notable rate acceleration due to the more enhanced ion-dipole interaction with the quinone carbonyl oxygen. Spectroscopic titration (absorption and (1)H NMR) and kinetic experiments indicated that only the longest di-armed 2c constructs 1:1, and then 1:2, host/guest complexes with Ca(2+), Sr(2+), and Ba(2+). These 1:2 complexes exhibited the most effective acceleration for the respective metal cations. PMID- 12733921 TI - Theoretical calculations of carbon-oxygen bond dissociation enthalpies of peroxyl radicals formed in the autoxidation of lipids. AB - Theoretical calculations were carried out to provide a framework for understanding the free radical oxidation of unsaturated lipids. The carbon[bond]hydrogen bond dissociation enthalpies (BDEs) of organic model compounds and oxidizable lipids (R[bond]H) and the carbon[bond]oxygen bond dissociation enthalpies of peroxyl radical intermediates (R[bond]OO*) have been calculated. The carbon[bond]hydrogen BDEs correlate with the rate constant for propagation of free radical autoxidation, and the carbon[bond]oxygen BDEs of peroxyl radicals correlate with rate constants for beta-fragmentation of these intermediates. Oxygen addition to intermediate carbon radicals apparently occurs preferentially at centers having the highest spin density. The calculated spin distribution therefore provides guidance about the partitioning of oxygen to delocalized carbon radicals. Where the C[bond]H BDEs are a function of the extent of conjugation in the parent lipid and the stability of the carbon radical derived therefrom, C[bond]OO* BDEs are also affected by hyperconjugation. This gives way to different rates of beta-fragmentation of peroxyl radicals formed from oxygen addition at different sites along the same delocalized radical. We have also studied by both theory and experiment the propensity for benzylic radicals to undergo oxygen addition at their ortho and para carbons which, combined, possess an equivalent unpaired electron spin density as the benzylic position itself. We find that the intermediate peroxyl radicals in these cases have negative C[bond]OO* BDEs and, thus, have rate constants for beta fragmentation that exceed the diffusion-controlled limit for the reaction of a carbon-centered radical with oxygen. PMID- 12733923 TI - Glycopeptide-membrane interactions: glycosyl enkephalin analogues adopt turn conformations by NMR and CD in amphipathic media. AB - Four enkephalin analogues (Tyr-D-Thr-Gly-Phe-Leu-Ser-CONH(2), 1, and the related O-linked glycopeptides bearing the monosaccharide beta-glucose, 2, the disaccharide beta-maltose, 3, and the trisaccharide beta-maltotriose, 4) were synthesized, purified by HPLC, and biophysical studies were conducted to examine their interactions with membrane model systems. Glycopeptide 2 has been previously reported to penetrate the blood-brain barrier (BBB), and produce potent analgesia superior to morphine in mice (J. Med. Chem.2000, 43, 2586-90 and J. Pharm. Exp. Ther. 2001, 299, 967-972). The parent peptide and its three glycopeptide derivatives were studied in aqueous solution and in the presence of micelles using 2-D NMR, CD, and molecular mechanics (Monte Carlo studies). Consistent with previous conformational studies on cyclic opioid agonist glycopeptides, it was seen that glycosylation did not significantly perturb the peptide backbone in aqueous solution, but all four compounds strongly associated with 5-30 mM SDS or DPC micelles, and underwent profound membrane-induced conformational changes. Interaction was also observed with POPC:POPE:cholesterol lipid vesicles (LUV) in equilibrium dialysis experiments. Although the peptide backbones of 1-4 possessed random coil structures in water, in the presence of the lipid phase they each formed a nearly identical pair of structures, all with a stable beta-turn motif at the C-terminus. Use of spin labels (Mn(2+) and 5 DOXYL-stearic acid) allowed for the determination of the position and orientation of the compounds relative to the surface of the micelle. PMID- 12733924 TI - A polyphenylene dendrimer-detergent complex as a highly fluorescent probe for bioassays. AB - The synthesis of a polyphenylene dendrimer carrying three perylenemonoimide dyes as well as one biotin group is presented. Due to the hydrophobic polyphenylene scaffold, this dendrimer is insoluble in water thus preventing investigations in aqueous media. However, the use of an appropriate detergent results in the formation of well-defined supramolecular dendrimer-detergent complexes being soluble in aqueous media. The dendrimer-detergent complexes have a constant hydrodynamic radius of 7.1 nm measured by light scattering and fluorescence correlation spectroscopy and exhibit a high stability in the presence of blood serum proteins. The specific binding of the dendrimer-detergent complexes carrying a single biotin group to the protein streptavidin is demonstrated using a magnetic bead assay. PMID- 12733925 TI - Altering the allowed/forbidden gap in cyclobutene electrocyclic reactions: experimental and theoretical evaluations of the effect of planarity constraints. AB - The allowed conrotatory cyclobutene ring-opening has a distinctly nonplanar carbon skeleton. Classic experiments by Brauman and Archie, and by Freedman et al., placed the allowed/forbidden gap at greater than 15 kcal/mol. Wolfgang Roth proposed that a system forced to planarity might have a smaller preference for the conrotatory mode than unconstrained systems. Such systems have now been studied theoretically and experimentally, and results that confirm Roth's postulate are presented here. The experiments were performed in Bochum, and the calculations were carried out in Osaka and Los Angeles. As the cyclobutene ring opening transition structure approaches planarity, the energy gap between allowed conrotatory and the forbidden disrotatory pathways decreases. For the ring opening of a cyclobutene fused to norbornene, the energy gap between the forbidden and the allowed transition state is only 4.1 kcal/mol by CASSCF and 8.0 kcal/mol by CAS-MP2 as compared to 13.4 and 19.2 kcal/mol, respectively, for the parent cyclobutene. Experimental studies of 3,4-dimethylcyclobutenes fused to various ring systems are reported, and a trend is found toward a reduced allowed/forbidden gap as the planarity of the cyclobutene is enforced. PMID- 12733926 TI - Excitation energy transport processes of porphyrin monomer, dimer, cyclic trimer, and hexamer probed by ultrafast fluorescence anisotropy decay. AB - Femtosecond fluorescence anisotropy measurements for a variety of cyclic porphyrin arrays such as Zn(II)porphyrin m-trimer and hexamer are reported along with o-dimer and monomer as reference molecules. In the porphyrin arrays, a pair of porphyrin moieties are joined together via triphenyl linkage to ensure cyclic and rigid structures. Anisotropy decay times of the porphyrin arrays can be well described by the Forster incoherent excitation hopping process between the porphyrin units. Exciton coupling strengths of 74 and 264 cm(-1) for the m-trimer and hexamer estimated from the observed excitation energy hopping rates are close to those of B800 and B850, respectively, in the LH2 bacterial light-harvesting antenna. Thus, these cyclic porphyrin array systems have proven to be useful in understanding energy migration processes in a relatively weak interaction regime in light of the similarity in overall structures and constituent chromophores to natural light-harvesting arrays. PMID- 12733927 TI - Fluorous catalysis under homogeneous conditions without fluorous solvents: a "greener" catalyst recycling protocol based upon temperature-dependent solubilities and liquid/solid phase separation. AB - The thermomorphic fluorous phosphines P((CH(2))(m)()(CF(2))(7)CF(3))(3) (m = 2, 1a; m = 3, 1b) exhibit ca. 600-fold solubility increases in n-octane between -20 (1a = 0.104 mM) and 80 degrees C (63.4 mM) and 1500-fold solubility increases between -20 and 100 degrees C (151 mM). They catalyze conjugate additions of alcohols to methyl propiolate under homogeneous conditions in n-octane at 65 degrees C and can be recovered by simple cooling and precipitation and used again. This avoids the use of fluorous solvents during the reaction or workup, which are expensive and can leach in small amounts. Teflon shavings can be used to mechanically facilitate recycling, and (31)P NMR analyses indicate >97% phosphorus recovery (85.2% 1a, 12.2% other). (19)F NMR analyses show that 2.3% of the (CF(2))(7)CF(3) moieties of 1a leach, in some form, into the n-octane (value normalized to phosphorus). 1a similarly catalyzes additions in the absence of solvent. Yield data match or exceed those of reactions conducted under fluorous/organic liquid/liquid biphase conditions. The extra methylene groups render 1b more nucleophilic than 1a and, thus, a more active catalyst. The temperature dependence of the solubility of 1a is measured in additional solvents and compared to that of the nonfluorous phosphine PPh(3). PMID- 12733929 TI - Molecular wires built from binuclear cyclometalated complexes. AB - Binuclear complexes with cyclometalated ends of the [Ru(bpy)(2)(ppH)](+) type (bpy = 2,2'-bipyridine, ppH = 2-phenylpyridine), linked by various spacers, have been prepared. These spacers are made of one or two triple bonds, or bis-ethynyl aryl groups, with aryl = benzene, thiophene, or anthracene. The complexes with bis-ethynyl aryl spacers are obtained by Sonogashira couplings with suitable bis alkynes, starting from the [Ru(bpy)(2)(ppBr)](+) synthon. Complexes with one or two triple bonds are obtained from the true alkyne [Ru(bpy)(2)(pp-CCH)](+) cyclometalated precursor, using respectively a Sonogashira coupling with the iodo derivative [Ru(bpy)(2)(ppI)](+), or an oxidative homocoupling. Some complexes with tert-butyl-substituted bipyridine ancillary ligands have also been obtained. Oxidation of the binuclear complexes occurs near 0.5 V, i.e., more easily than with [Ru(bpy)(3)](2+)-based complexes. A single anodic wave is observed, with almost no detectable splitting, corresponding to two closely spaced one-electron processes. Differential pulse voltammetry allows the determination of the corresponding comproportionation constants involving the mixed valence Ru(II)[bond]Ru(III) forms. Controlled potential electrolysis yields the mixed valence forms in comproportionation equilibrium with homovalent forms. Analysis of the intervalence transitions allows the calculation of the electronic coupling element V(ab). This series of complexes exhibit relatively large couplings when comparing with complexes of similar metal-metal distances, with a special mention for the anthracene-containing spacer, which appears particularly efficient for mediating the metal-metal interaction. The results can be rationalized by theoretical calculations at the extended Huckel level. PMID- 12733928 TI - Au nanocrystal growth on nanotubes controlled by conformations and charges of sequenced peptide templates. AB - A new biological approach to fabricate Au nanowires was examined by using sequenced peptide nanotubes as templates. The sequenced histidine-rich peptide molecules were assembled on nanotubes, and the biological recognition of the sequenced peptide selectively trapped Au ions for the nucleation of Au nanocrystals. After Au ions were reduced, highly monodisperse Au nanocrystals were grown on nanotubes. The conformations and the charge distributions of the histidine-rich peptide, determined by pH and Au ion concentration in the growth solution, control the size and the packing density of Au nanocrystals. The diameter of Au nanocrystal was limited by the spacing between the neighboring histidine-rich peptides on nanotubes. A series of TEM images of Au nanocrystals on nanotubes in the shorter Au ion incubation time periods reveal that Au nanocrystals grow inside the nanotubes first and then cover the outer surfaces of nanotubes. Therefore, multiple materials will be coated inside and outside the nanotubes respectively by controlling doping ion concentrations and their deposition sequences. It should be noted that metallic nanocrystals in diameter around 6 nm are in the size domain to observe a significant conductivity change by changing the packing density, and therefore this system may be developed into a conductivity-tunable building block. PMID- 12733930 TI - Adsorption of CF4 on the internal and external surfaces of opened single-walled carbon nanotubes: a vibrational spectroscopy study. AB - Infrared spectroscopy has been used to make the first experimental discrimination between molecules bound by physisorption on the exterior surface of carbon single walled nanotubes (SWNTs) and molecules bound in the interior. In addition, the selective displacement of the internally bound molecules has been observed as a second adsorbate is added. SWNTs were opened by oxidative treatment with O(3) at room temperature, followed by heating in a vacuum to 873 K. It was found that, at 133 K and 0.033 Torr, CF(4) adsorbs on closed SWNTs, exhibiting its nu(3) asymmetric stretching mode at 1267 cm(-1) (red shift relative to the gas phase, 15 cm(-1)). Adsorption on the nanotube exterior is accompanied by adsorption in the interior in the case of opened SWNTs. Internally bound CF(4) exhibits its nu(3) mode at 1247 cm(-1) (red shift relative to the gas phase, 35 cm(-1)). It was shown that, at 133 K, Xe preferentially displaces internally bound CF(4) species, and this counterintuitive observation was confirmed by molecular simulations. The confinement of CF(4) inside (10,10) single-walled carbon nanotubes does not result in the production of lattice modes that are observed in large 3D ensembles of CF(4). PMID- 12733931 TI - Spectroscopic and computational studies of Co3+-corrinoids: spectral and electronic properties of the B12 cofactors and biologically relevant precursors. AB - The B(12) cofactors methylcobalamin (MeCbl) and 5'-deoxyadenosylcobalamin (AdoCbl) have long fascinated chemists because of their complex structures and unusual reactivities in biological systems; however, their electronic absorption (Abs) spectra have remained largely unassigned. In this study, we have used Abs, circular dichroism (CD), magnetic CD (MCD), and resonance Raman spectroscopic techniques to probe the electronic excited states of Co(3+)Cbl species that differ with respect to their upper axial ligand, including MeCbl, AdoCbl, aquacobalamin (H(2)OCbl(+)), and vitamin B(12) (cyanocobalamin, CNCbl). Also included to probe the effect of the lower axial ligand on the electronic properties of Cbls is Ado-cobinamide (AdoCbi(+)), an AdoCbl derivative that lacks the tethered base 5,6-dimethylbenzimidazole (DMB) and instead binds a water molecule in the lower axial position. Spectroscopic data for each species are analyzed within the framework of time-dependent density functional theory (TD DFT) to assign the major spectral features (the so-called alpha/beta, D/E, and gamma bands) and to generate experimentally validated electronic-structure descriptions. These studies reveal that the "unique" Abs spectra of MeCbl and AdoCbl, which differ considerably from the "typical" Abs spectra of H(2)OCbl(+) and CNCbl, reflect the high degree of sigma-donation from the alkyl ligand to the Co center and the consequent destabilization of all Co 3d orbitals. They reveal further that with increasing sigma-donor strength of the upper axial ligand, the contribution from the formally unoccupied Co 3d(z(2)) orbital to the HOMO increases, which induces a strong Co[bond]N(DMB) sigma-antibonding interaction, consistent with the experimentally observed lengthening of this bond from H(2)OCbl(+) to CNCbl and MeCbl. Alternatively, our spectroscopic and computational data for MeCbl and MeCbi(+) reveal that substitution of the DMB by a water molecule in the lower axial position has negligible effects on the Co[bond]C. A simple model is presented that explains why the identity of the upper axial ligand has a major effect on the Co[bond]N(ax) strength, whereas the lower axial ligand does not appreciably modulate the nature of the Co[bond]C. Implications of these results with respect to enzymatic Co[bond]C activation are discussed. PMID- 12733932 TI - Axial solvent coordination in "base-fff" cob(II)alamin and related co(II) corrinates revealed by 2D-EPR. AB - Detailed information on the structure of cobalt(II) corrinates is of interest in the context of studies on the coenzyme B(12) catalyzed enzymatic reactions, where cob(II)alamin has been identified as a reaction intermediate. Cob(II)ester (heptamethyl cobyrinate perchlorate) is found to be soluble in both polar and nonpolar solvents and is therefore very suitable to study solvent effects on Co(II) corrinates. In the literature, Co(II) corrinates in solution are often addressed as four-coordinated Co(II) corrins. However, using a combination of continuous-wave (CW) and pulse electron paramagnetic resonance (EPR) and pulse ENDOR (electron nuclear double resonance) at different microwave frequencies we clearly prove axial ligation for Cob(II)ester and the base-off form of cob(II)alamin (B(12r)) in different solvents. This goal is achieved by the analysis of the g values, and the hyperfine couplings of cobalt, some corrin nitrogens and hydrogens, and solvent protons. These parameters are shown to be very sensitive to changes in the solvent ligation. Density functional computations (DFT) facilitate largely the interpretation of the EPR data. In the CW-EPR spectrum of Cob(II)ester in methanol, a second component appears below 100 K. Different cooling experiments suggest that this observation is related to the phase transition of methanol from the alpha-phase to the glassy state. A detailed analysis of the EPR parameters indicates that this transition induces a change from a five-coordinated (above 100 K) to a six-coordinated (below 100 K) Co(II) corrin. In a CH(3)OH:H(2)O mixture the phase-transition properties alter and only the five-coordinated form is detected for Cob(II)ester and for base-off B(12r) at all temperatures. Our study thus shows that the characteristics of the solvent can have a large influence on the structure of Co(II) corrinates and that comparison with the protein-embedded cofactor requires some caution. Finally, the spectral similarities between Cob(II)ester and base-off B(12r) prove the analogies in their electronic structure. PMID- 12733933 TI - 51V magic angle spinning NMR spectroscopy of six-coordinate Lindqvist oxoanions: a sensitive probe for the electronic environment in vanadium-containing polyoxometalates. Counterions dictate the 51V fine structure constants in polyoxometalate solids. AB - Geometric and electronic environments of vanadium have been addressed by (51)V magic angle spinning NMR spectroscopy of six-coordinated polyoxometalate solids. (C(4)H(9))(4)N(+) and mixed Na(+)/Cs(+) salts of the Lindqvist-type mono- and divanadium-substituted oxotungstates, [VW(5)O(19)](3-) and [V(2)W(4)O(19)](4-), have been prepared as microcrystalline and crystalline solids. The solid-state NMR spectra reflect the details of the local environment of the vanadium site in these hexametalate solids via the anisotropic quadrupolar and chemical shielding interactions. Remarkably, these (51)V fine structure constants in the solid state are dictated by the nature and geometry of the countercations. Electrostatic calculations of the electric field gradients at the vanadium atoms have been performed. Experimental trends are well reproduced with the simple electrostatic model, and explain the sensitivity of the anisotropic NMR parameters to the changes in the cationic environment at the vanadium site. PMID- 12733934 TI - Contributions of symmetric and asymmetric normal coordinates to the intervalence electronic absorption and resonance Raman spectra of a strongly coupled p phenylenediamine radical cation. AB - Resonance Raman spectroscopy, electronic absorption spectroscopy, and the time dependent theory of spectroscopy are used to analyze the intervalence electron transfer properties of a strongly delocalized class III molecule, the tetraalkyl p-phenylene diamine radical cation bis(3-oxo-9-azabicyclo[3.3.1]non-9-yl)benzene ((k33)(2)PD(+)). This molecule is a prototypical system for strongly coupled organic intervalence electron transfer spectroscopy. Resonance Raman excitation profiles in resonance with the lowest energy absorption band are measured. The normal modes of vibration that are most strongly coupled to the intervalence transition are identified and assigned by using UB3LYP/6-31G(d) calculations. Excited state distortions are obtained, and the resonance Raman intensities and excitation profiles are calculated by using the time-dependent theory of Raman spectroscopy. The most highly distorted normal modes are all totally symmetric, but intervalence electron transfer absorption spectra are usually interpreted in terms of a model based on coupling between potential surfaces that are displaced along an asymmetric normal coordinate. This model provides a convenient physical picture for the intervalence compound, but it is inadequate for explaining the spectra. The absorption spectrum arising from only the strongly coupled surfaces consists of a single narrow band, in contrast to the broad, vibronically structured experimental spectrum. The electronic absorption spectrum of (k33)(2)PD(+) is calculated by using exactly the same potential surfaces as those used for the Raman calculations. The importance of symmetric normal coordinates, in addition to the asymmetric coordinate, is discussed. The observed vibronic structure is an example of the missing mode effect; the spacing is interpreted in terms of the time-dependent overlaps in the time domain. PMID- 12733935 TI - Long-range electron transfer through monolayers and bilayers of alkanethiols in electrochemically controlled Hg[bond]neling junctions. AB - The rates of electron tunneling through monolayers and bilayers of alkanethiols self-assembled in a potentiostatically controlled Hg-Hg junction are reported. An alkanethiolate monolayer is formed in situ on one or both Hg drops via oxidative adsorption at the controlled potential. Subsequently, the Hg drops are brought into contact using micromanipulators. The junction formation is instantly followed by the flow of a steady-state tunneling current between the two electrodes. A plot of the logarithm of the tunneling current density vs the total number of carbon atoms in each junction yields identical tunneling coefficients, beta = 1.06 +/- 0.04/-CH(2)- and beta = 1.02 +/- 0.07/-CH(2)-, for monolayers and bilayers of alkanethiols, respectively. Careful examination of the tunneling data indicates that the solvent and ions are ejected from the junction area. The tunneling current recorded for a bilayer of 1-octanethiol or 1-nonanethiol is ca. 2-fold larger than a corresponding tunneling current recorded for monolayers of 1 hexadecanethiol or 1-octadecanethiol, respectively. This result is explained in terms of weak electronic coupling across the noncovalent molecule/electrode interface. PMID- 12733937 TI - Comparison of formation of reactive conformers (NACs) for the Claisen rearrangement of chorismate to prephenate in water and in the E. coli mutase: the efficiency of the enzyme catalysis. AB - The Claisen rearrangements of chorismate (CHOR) in water and at the active site of E. coli chorismate mutase (EcCM) have been compared. From a total of 33 ns molecular dynamics simulation of chorismate in water solvent, seven diaxial conformers I-VII were identified. Most of the time (approximately 99%), the side chain carboxylate of the chorismate is positioned away from the ring due to the electrostatic repulsion from the carboxylate in the ring. Proximity of the two carboxylates, as seen in conformer I, is a requirement for the formation of a near attack conformer (NAC) that can proceed to the transition state (TS). In the EcCM.CHOR complex, the two carboxylates of CHOR are tightly held by Arg28 of one subunit and Arg11* of the other subunit, resulting in the side chain C16 being positioned adjacent to C5 with their motions restricted by van der Waals contacts with methyl groups of Val35 and Ile81. With the definition of NAC as the C5...C16 distance < or =3.7 A and the attack angle < or =30 degrees, it was estimated from our MD trajectories that the free energy of NAC formation is approximately 8.4 kcal/mol above the total ground state in water, whereas in the enzyme it is only 0.6 kcal/mol above the average of the Michaelis complex EcCM.CHOR. The experimentally measured difference in the activation free energies of the water and enzymatic reactions (Delta Delta G(++)) is 9 kcal/mol. It follows that the efficiency of formation of NAC (7.8 kcal/mol) at the active site provides approximately 90% of the kinetic advantage of the enzymatic reaction as compared to the water reaction. Comparison of the EcCM.TSA (transition state analogue) and EcCM.NAC simulations suggests that the experimentally measured 100 fold tighter binding of TSA compared to CHOR does not originate from the difference between NAC and the TS binding affinities, but might be due to the free energy cost to bring the two carboxylates of CHOR together to interact with Arg28 and Arg11* at the active site. The two carboxylates of TSA are fixed by a bicyclic structure. The remaining approximately 10% of Delta Delta G(++) may be attributed to a preferential interaction of Lys39-NH(3)(+) with O13 ether oxygen in the TS. PMID- 12733938 TI - Electronic basis of improper hydrogen bonding: a subtle balance of hyperconjugation and rehybridization. AB - The X[bond]H bond length in X[bond]H...Y hydrogen bonded complexes is controlled by a balance of two main factors acting in opposite directions. "X[bond]H bond lengthening" due to n(Y)-->sigma(H[bond]X) hyperconjugative interaction is balanced by "X[bond]H bond shortening" due to increase in the s-character and polarization of the X[bond]H. When hyperconjugation dominates, X[bond]H bond elongation is reflected in a concomitant red shift of the corresponding IR stretching frequency. When the hyperconjugative interaction is weak and the X hybrid orbital in the X[bond]H is able to undergo a sufficient change in hybridization and polarization, rehybridization dominates leading to a shortening of the X[bond]H and a blue shift in the X[bond]H stretching frequency. PMID- 12733936 TI - N[bond]C(alpha) bond dissociation energies and kinetics in amide and peptide radicals. Is the dissociation a non-ergodic process? AB - Dissociations of aminoketyl radicals and cation radicals derived from beta alanine N-methylamide, N-acetyl-1,2-diaminoethane, N(alpha)-acetyl lysine amide, and N(alpha)-glycyl glycine amide are investigated by combined density functional theory and Moller-Plesset perturbational calculations with the goal of elucidating the mechanism of electron capture dissociation (ECD) of larger peptide and protein ions. The activation energies for dissociations of N[bond]C bonds in aminoketyl radicals decrease in the series N[bond]CH(3) > N CH(2)CH(2)NH(2) >> N[bond]CH(2)CONH(2) approximately N[bond]CH(CONH(2))(CH(2))(4)NH(2). Transition state theory rate constants for dissociations of N[bond]C(alpha) bonds in aminoketyl radicals and cation-radicals indicate an extremely facile reaction that occurs with unimolecular rate constants >10(5) s(-1) in species thermalized at 298 K in the gas phase. In neutral aminoketyl radicals the N[bond]C(alpha) bond cleavage results in fast dissociation. In contrast, N[bond]C(alpha) bond cleavage in aminoketyl cation radicals results in isomerization to ion-molecule complexes that are held together by strong hydrogen bonds. The facile N[bond]C(alpha) bond dissociation in thermalized ions indicates that it is unnecessary to invoke the hypothesis of non-ergodic behavior for ECD intermediates. PMID- 12733939 TI - Interference between the hydrogen bonds to the two rings of nicotine. AB - The biochemical transport and binding of nicotine depends on the hydrogen bonding between water and binding site residues to the pyridine ring and the protonated pyrrolidinium ring. To test the independence of these two moderately separated hydrogen-bonding sites, we have calculated the structures of clusters of protonated nicotine with water and a bicarbonate anion, benzene, indole, or a second water molecule. Unprotonated nicotine-water clusters have also been studied for contrast. The potential energy surfaces are first explored with an intermolecular anisotropic atom-atom model potential. Full geometry optimizations are then carried out using density functional theory to include nonadditive terms in the interaction energies. The presence of the charge on the pyrrolidine nitrogen removes the conventional hydrogen-bonding site on the pyridine ring. The hydrogen-bond ability of this site is nearly recovered when the protonated pyrrolidinium ring is bound to a bicarbonate anion, whereas its interaction with benzene shows a much smaller effect. Indole appears to partially restore the hydrogen-bond ability of the pyridine nitrogen, although indole and benzene both pi-bond to the pyrrolidinium ring. A second hydrogen-bonding water produces a significant conformational distortion of the nicotine. This demonstrates the limitations of the conventional qualitative predictions of hydrogen bonding based on the independence of molecular fragments. It also provides benchmarks for the development of atomistic modeling of biochemical systems. PMID- 12733940 TI - Solution-processed anodes from layer-structure materials for high-efficiency polymer light-emitting diodes. AB - The development of low-cost, large-area electronic applications requires the deposition of active materials in simple and inexpensive techniques at room temperature, properties usually associated with polymer films. In this study, we demonstrate the integration of solution-processed inorganic films in light emitting diodes. The layered transition metal dichalcogenide (LTMDC) films are deposited through Li intercalation and exfoliation in aqueous solution and partially oxidized in an oxygen plasma generator. The chemical composition and thickness of the LTMDC and corresponding transition metal oxide (TMO) films are investigated by X-ray photoelectron spectroscopy. The morphology and topography of the films are studied by atomic force microscopy. X-ray powder diffraction is used to determine the orientation of the LTMDC film. Finally, the LTMDC and their corresponding oxides are utilized as hole-injecting and electron-blocking materials in polymer light-emitting diodes with the general structure ITO/LTMDC/TMO/polyfluorene/Ca/Al. Efficient hole injection and electron blocking by the inorganic layers result in outstanding device performance and high efficiency. PMID- 12733949 TI - Delivery of transforming growth factor-beta2-perturbing antibody in a collagen vehicle inhibits cranial suture fusion in calvarial organ culture. AB - OBJECTIVE: To determine whether antibody perturbation of Tgf-beta, delivered in a collagen gel, could inhibit cranial suture fusion. DESIGN: Attachment and proliferation of osteoblasts cultured on a collagen gel with or without anti-Tgf beta2 antibody were determined by AlamarBlue dye assay and cell morphology by toluidine-blue staining. In rat calvarial organ culture, collagen gel with and without anti-Tgf-beta2 antibody was injected subperiosteally over the posterior frontal suture of postnatal day 15 rat calvariae. A quantitative analysis of suture fusion was used to measure suture bridging in histological serial sections at various time points. RESULTS: Attachment and proliferation for cells cultured on collagen gel with anti-Tgf-beta2 antibody were similar to collagen gel controls. Although proliferation was lower than on tissue culture plastic, cells treated with anti-Tgf-beta2 antibody maintained an osteoblastic morphology. After 7, 10, and 15 days in organ culture, anti-Tgf-beta2 antibody treatment caused a reduction in the percent bridging of posterior frontal sutures, compared with controls. Sutures exposed to anti-Tgf-beta2 antibody and fibroblast growth factor 2 concurrently did not show an inhibition of bony bridging. CONCLUSIONS: These results support previous reports suggesting a role for Tgf-beta2 in cranial suture fusion. In cell culture the collagen gel, both with and without anti-Tgf beta2 antibody, promoted similar osteoblast attachment, proliferation, and osteoblastic morphology. In organ culture anti-Tgf-beta2 antibody was delivered in a bioactive state via a collagen gel to inhibit cranial suture fusion. Also, the results suggest that the inductive effect of fibroblast growth factor-2 is not dependent on Tgf-beta2 activity. Together, these results provide further support for the role of Tgf-beta2 in cranial suture fusion. PMID- 12733950 TI - Cephalometric analysis of the consolidation phase following bilateral pediatric mandibular distraction. AB - OBJECTIVE: The goal of the consolidation phase of mandible distraction is to maintain the improvement in maxillomandibular form and relationship while the generated tissue ossifies. During this period, external deforming forces can act on the healing generated bone. The purpose of this study was to describe the potential cephalometric changes that occur following pediatric bilateral mandibular distraction using external devices. DESIGN: Retrospective lateral superimposition cephalometric analyses. PARTICIPANTS: Thirty-five cases of pediatric mandible distraction were reviewed. Seven of these cases were included in the study after exclusion criteria were applied. These cases represented a group with severe congenital dysmorphology and a mean device activation of 26.5 mm. MAIN OUTCOME MEASURES: Changes in pogonion position, symphyseal plane rotation, mandible length, and mandible length relative to maxillary length during the 18 to 36 days of activation, the eight weeks of consolidation, and the 1-year period following removal of the distraction device were measured. RESULTS: All patients demonstrated variable changes in position of the mandible during the consolidation phase. The most common were retrusion of pogonion, a decrease in mandible length, and a clockwise rotation of the symphyseal plane. In some cases the changes that occurred during consolidation were greater than those that occurred on 1-year follow-up. CONCLUSIONS: The consolidation phase of distraction osteogenesis is a dynamic phase and should not be assumed to be static. Multicenter use of this cephalometric technique would help to identify potential risk factors associated with postactivation changes. PMID- 12733953 TI - Single motor unit activity in levator veli palatini during speech and nonspeech tasks. AB - OBJECTIVE: This article assesses the control of velar movement by relating observed recruitment patterns of single motor unit activity in levator veli palatini observed during speech and nonspeech tasks in a single subject to intraoral pressure demands. METHODS: Electromyographic activity was recorded from a single motor unit in levator veli palatini during repetitions of "Say (----) again" with selected consonant-vowel-consonant and consonant-vowel syllables, sustained high pressure consonants, and blowing tasks. Single motor unit firing characteristics (e.g., frequency of occurrence, firing frequency) were related to intraoral air pressures recorded during the sustained consonant and blowing tasks. RESULTS: Levator single motor unit activity was always present during the /s/ in "say" and the first and second /s/ in /sis/. Activity was observed less consistently during the production of the /s/ in /sus/, the /p/ in /p Lambda/, and the /g/ in "again." Single motor unit firing frequency ranged from 16.1 Hz to 22 Hz during phrase productions. Recruitment was observed during sustained productions of high-pressure consonants when intraoral pressures exceeded 15 cm H(2)O. Increases in intraoral air pressure were associated with 25% to 85% increases in single motor unit firing frequencies. During nonspeech blowing tasks, single motor unit activity was observed when intraoral air pressure exceeded approximately 12 cm H(2)O. Increases in intraoral air pressure were again associated with increased single motor unit firing rates. CONCLUSIONS: Results showed evidence of both preprogrammed and feedback-controlled responses by levator veli palatini to changes in task intraoral pressure demands. PMID- 12733952 TI - Cephalometric and occlusal outcome in adults with unilateral cleft lip, palate, and alveolus after two different surgical techniques. AB - OBJECTIVE: To assess differences in the aesthetic and functional long-term results of one-stage and two-stage surgical and orthodontic treatment in patients with cleft lip, palate, and alveolus. DESIGN: Sixty adult patients who were operated on as children for unilateral cleft lip, palate, and alveolus were examined. In every patient the lip was closed using Tennison's technique. Thirty patients had soft and hard palate closure in two stages and 30 patients in a single stage. Lateral cephalometric and model analyses were conducted at a mean age of 18.4 years. RESULTS: In the model analysis, transverse narrowing was seen in all patients after two-stage operations and in three patients after one-stage operations. The deficit was more severe in the molar region in the two-stage group and nearly similar in the premolar and molar region in the one-stage group. A sagittal deficiency in the anterior maxilla was found in 26 patients after two stage operations and in 16 patients after one-stage operations. In the lateral cephalometric analysis, the mean sella-nasion-point A angle in the one- and two stage group was 78.2 degrees and 76.8 degrees, respectively. The ANB angle was normal in both groups. In both groups the inclination of the midface was low. There was a low posterior facial height. Minor scarring was seen in the single stage group. CONCLUSION: A more severe impairment of growth of the maxilla in the sagittal and frontal plane was observed after two-stage operations on the cleft palate. PMID- 12733951 TI - Association between subjective and objective measures of lip form and function: an exploratory analysis. AB - OBJECTIVE: In patients with cleft lip and palate, the aims of this study were to generate objective measures of different attributes of lip movement and to explore the utility of these objective measures by examining the association between examiners' subjective assessments with the objective measures. PATIENTS AND PARTICIPANTS: Thirteen patients with unilateral cleft lip and palate with varying degrees of cleft scar severity were selected. All patients had a previously repaired complete unilateral cleft lip and palate. INTERVENTIONS: Photographs and videotape recordings were made of the patients with cleft at rest and during smiling. Measurements of lip movement were obtained by means of a motion analysis system. MAIN OUTCOME MEASURES: The study sought to obtain rankings of cleft scar severity and impairment on a 6-point Likert scale by a lay and professional panel and measurements of displacement, asymmetry, speed, and velocity of upper lip during smiling. RESULTS: Displacement was the most consistent and valid objective measurement. An objective analysis of the entire upper lip provided the most information. In general, there was a decrease in the objective measures of upper lip movement as examiners' perceptions of facial appearance or disfigurement at rest and impairment during movement became worse. This relationship was stronger for the at-rest perceptions, implying that subjective assessments should be made with the face at rest. CONCLUSIONS: Objective measures provided the promise for differentiation of the components of movement and should be used to supplement subjective evaluations of lip appearance at rest and during movement. PMID- 12733954 TI - Unilateral Moore pharyngoplasty in the treatment of unilateral or asymmetric velopharyngeal incompetence. AB - OBJECTIVE: To describe the use of a unilateral modification of the Moore pharyngoplasty in the treatment of unilateral or asymmetric velopharyngeal incompetence (VPI) and analyze the results in a consecutive series of patients operated on by a single surgeon. DESIGN: Blind assessment of randomized speech and nasendoscopy recordings. SETTING: A two-site tertiary referral cleft unit. PATIENTS: Eighteen consecutive patients with asymmetrical or unilateral VPI of varying etiology. INTERVENTIONS: A unilateral Moore pharyngoplasty was performed in all patients. Three patients underwent radical dissection and retropositioning of the velar muscles at the same time as the unilateral Moore pharyngoplasty. MAIN OUTCOME MEASURES: Pre- and postoperative nasality and nasal airflow using the CAPS score, assessment of nasendoscopy recordings, and the rate of further surgery. RESULTS: There was a significant improvement in hypernasality (p =.014). There was a highly significant decrease in the size of the velopharyngeal gap on the side on which the Moore pharyngoplasty was performed (p =.004) as well as a highly significant decrease in the total gap size (p =.003). The Moore flap was effective in obliterating the lateral pharyngeal recess in 11 of 12 patients (p =.004). Three patients required further velopharyngeal surgery. CONCLUSIONS: In appropriately selected patients, a unilateral Moore pharyngoplasty is a safe and effective treatment for unilateral or asymmetric VPI. If indicated, a radical dissection and retropositioning of the velar muscles may be combined with a Moore pharyngoplasty. PMID- 12733955 TI - Longitudinal follow-up of obstructive sleep apnea following Furlow palatoplasty in children with cleft palate: a preliminary report. AB - OBJECTIVE: To longitudinally investigate the incidence and severity of obstructive sleep apnea (OSA) following Furlow palatoplasty for velopharyngeal insufficiency (VPI) in children with cleft palate. SUBJECTS: Ten children, six boys and four girls, mean age 5.1 years, at Furlow palatoplasty. DESIGN: Prospective analysis. MAIN OUTCOME MEASURES: Overnight polysomnographic studies were used to determine the incidence and severity of sleep apneas 1 day prior to Furlow palatoplasty, 1 week postoperatively, and approximately 3 and 6 months postoperatively. RESULTS: None of the patients suffered OSA prior to Furlow palatoplasty. A high incidence of mild OSA (100%) occurred during the early postoperative period (p <.001) but resolved within 3 months in all but two patients (20%). Only one OSA (10%) persisted 6 months postoperatively. CONCLUSIONS: Furlow palatoplasty for VPI in children with cleft palate might induce temporary and mild OSA. PMID- 12733957 TI - Increased fistula risk following palatoplasty in Treacher Collins syndrome. AB - OBJECTIVE: Patients with Treacher Collins syndrome have abnormal vascular supply to the palate, yet it is unknown whether there are increased postoperative healing problems following palatoplasty. This study investigated the correlation between Treacher Collins syndrome and postoperative palatal fistula formation. DESIGN: Retrospective chart review was performed. PATIENTS: Children undergoing palatoplasty at Children's Hospital Los Angeles from 1987 to 2000 were evaluated. Ten children with Treacher Collins syndrome, 92 children with other syndromes and cleft palate, and 458 nonsyndromic patients with isolated cleft palate were studied. INTERVENTIONS: All children were treated with a one-stage, double reversing Z-plasty cleft palate repair. MAIN OUTCOME MEASURES: Outcome measures included intraoperative observations of surgical anatomy and postoperative clinic follow-up of fistula formation. Palatal fistula rates between patients with Treacher Collins syndrome, other syndromes, and no syndrome were compared with chi-square analysis. RESULTS: Children with Treacher Collins syndrome had significantly greater palatal fistula rates (50%) than children with other syndromes (8.7%) or no syndrome (4.1%). Treacher Collins patients demonstrated large palatal fistulas and poor flap vascularity. CONCLUSIONS: Children with Treacher Collins syndrome and cleft palate have significantly higher palatal fistula risk than other children with cleft palate when double-reversing Z-plasty palate repair is performed. Our findings suggest that children with Treacher Collins syndrome and cleft palate may have poor vascularity to palatal flaps created during palatoplasty. Furthermore, we recommend that surgeons performing palatoplasty minimize the dissection of mucoperiosteal flaps around the greater palatine arterial pedicle and utilize closure techniques creating the least vascular disruption of palatal tissue. PMID- 12733958 TI - Negative transcriptional regulation of connexin 43 by Tbx2 in rat immature coronal sutures and ROS 17/2.8 cells in culture. AB - BACKGROUND: Tbx2 is a member of the T-box family of transcriptional regulatory genes with an extensive but not yet fully understood role in embryonic development. This study explores the potential role of Tbx2 in calvarial morphogenesis. OBJECTIVES: To explore the hypothesis that Tbx2 has a negative regulatory effect on the expression of connexin 43 (Cx43), a protein necessary for cell-to-cell communication; document the presence of Tbx2 protein in the developing cranial sutures; and determine the spatial pattern of expression of this developmentally regulated transcription factor in calvariae. DESIGN: The osteoblast-like cell line ROS 17/2.8 was stably transfected with sense or antisense Tbx2. Immunohistochemistry and Western blotting was used to study Tbx2 and Cx43 expression in these cells and sections of embedded developing coronal sutures. RESULTS: The ROS 17/2.8 cells transfected with antisense Tbx2 showed a decrease in expression of Tbx2 protein and an increase in expression of endogenous Cx43. The reverse is seen with sense-transfected cells. Both of these proteins are expressed in rat developing coronal sutures. The pattern of Tbx2 expression in the developing was also reciprocal to the pattern of Cx43 expression. Tbx2 protein is concentrated in the center of the sutural blastema, an area devoid of Cx43 protein localization. Conversely, Tbx2 protein expression is low in the periphery of the sutures, in which there is high Cx43 protein expression. CONCLUSIONS: Taken together, these studies suggest that Tbx2 protein is a negative regulator of Cx43 expression at the transcriptional level in cranial sutures in vivo. PMID- 12733959 TI - Are facial expressions reproducible? AB - OBJECTIVES: To determine the extent of reproducibility of five facial expressions. DESIGN: Thirty healthy Caucasian volunteers (15 males, 15 females) aged 21 to 30 years had 20 landmarks highlighted on the face with a fine eyeliner pencil. Subjects were asked to perform a sequence of five facial expressions that were captured by a three-dimensional camera system. Each expression was repeated after 15 minutes to investigate intrasession expression reproducibility. To investigate intersession expression reproducibility, each subject returned 2 weeks after the first session. A single operator identified 3-dimensional coordinate values of each landmark. A partial ordinary procrustes analysis was used to adjust for differences in head posture between similar expressions. Statistical analysis was undertaken using analysis of variance (linear mixed effects model). RESULTS: Intrasession expression reproducibility was least between cheek puffs (1.12 mm) and greatest between rest positions (0.74 mm). The reproducibility of individual landmarks was expression specific. Except for the lip purse, the reproducibility of facial expressions was not statistically different within each of the two sessions. Rest position was most reproducible, followed by lip purse, maximal smile, natural smile, and cheek puff. Subjects did not perform expressions with the same degree of symmetry on each occasion. Female subjects demonstrated significantly better reproducibility with regard to the maximal smile than males (p =.036). CONCLUSIONS: Under standardized conditions, intrasession expression reproducibility was high. Variation in expression reproducibility between sessions was minimal. The extent of reproducibility is expression specific. Differences in expression reproducibility exist between males and females. PMID- 12733956 TI - Genetic association studies of cleft lip and/or palate with hypodontia outside the cleft region. AB - OBJECTIVE: The purpose of this study was to determine whether the candidate genes previously studied in subjects with cleft lip, cleft palate, or both are associated with hypodontia outside the region of the cleft. SUBJECTS: One hundred twenty subjects from the Iowa Craniofacial Anomalies Research Center were selected based on the availability of both dental records and genotype information. METHOD: The type of orofacial clefting and type and location of dental anomalies (missing teeth, supernumerary teeth, or peg laterals) were assessed by dental chart review and radiographic examination. Genotype analysis of candidate genes was performed using polymerase chain reaction/single-strand conformation polymorphism analysis. RESULTS: The prevalence of hypodontia in this sample was 47.5%, with 30.0% of subjects having missing teeth outside the cleft. There was a positive association between subjects with cleft lip or cleft lip and palate who had hypodontia outside the cleft region (compared with noncleft controls) and both muscle segment homeo box homolog 1 (MSX1) (p =.029) and transforming growth factor beta 3 (TGFB3) (p =.024). It was not possible in this analysis to determine whether this association was specifically associated with orofacial clefting combined with hypodontia or whether it was due primarily to the clefting phenotype. CONCLUSIONS: In this sample, there was a significantly greater incidence of hypodontia outside the cleft region in subjects with cleft lip and palate, compared with cleft lip only or cleft palate only. Cleft lip and/or palate with hypodontia outside the cleft region was positively associated with both TGFB3 and MSX1, compared with noncleft controls. PMID- 12733960 TI - An investigation of the relationship between associated congenital malformations and the mental and psychomotor development of children with clefts. AB - OBJECTIVE: This research studied the relationship between associated congenital malformations and the mental and psychomotor development of children with clefts. DESIGN: The study was cross-sectional. SETTING: The study was conducted in a university hospital for children. PARTICIPANTS: The sample consisted of 148 children with cleft lip, cleft palate, or both. MAIN OUTCOME MEASURES: The children were assessed by a clinical geneticist at the age of 18 months. The children's level of development was determined by means of the Dutch version of the Bayley Scales of Infant Development. RESULTS: One-third of the total sample had associated malformations. Children with an isolated cleft lip showed the least. Children with an isolated cleft palate showed the highest percentage of minor malformations that are minor yet possibly worrisome. The total group achieved a mean developmental index (DI) on the mental scale of 98.9 with SD of 20.9. The motor scale showed a mean DI of 104.9 and SD of 24.7. Analysis of variance (ANOVA) showed that on the mental scale, the three main effects (diagnosis, evaluation, and sex) were significant at the 5% level. On the motor scale, only the main effect "evaluation" was significant. CONCLUSIONS: This study demonstrated that children with associated congenital malformations might be disadvantaged with respect to their development. These malformations occurred most frequently with the cleft lip and palate and cleft palate only subgroups. More research, especially concerning the cleft palate only subgroup is needed because they are most at risk. PMID- 12733961 TI - Audiologic and tympanometric findings in children with cleft lip and palate. AB - OBJECTIVE: To evaluate the otologic and audiologic status of 50 children with repaired cleft lip, cleft palate, or both in Hacettepe University, Ankara, Turkey. DESIGN: Audiometric and tympanometric evaluation of 100 ears in 50 children were performed. Hearing levels < or = 15 dB and middle ear pressures between -50 to +50 decaPascals were considered to be normal. Results were examined according to cleft type and laterality. The least and most affected frequencies were calculated. A simple evaluation of speech characteristics including nasal resonance, nasal air escape, and errors of articulation was also performed. RESULTS: Sixty-three of the 100 ears had normal hearing status, whereas 40 had normal middle ear pressures. No evidence was found to suggest that individual cleft type and laterality of the ear had any effect on hearing loss or middle ear disease. Two-thirds of the patients had normal or acceptable degree of language skills. CONCLUSION: The final hearing status of patients with cleft palate is a result of a combination of surgical correction, developmental factors, and treatment of middle ear disease. Early and aggressive ventilation tube placement is the standard of cleft care in many countries. Our long-term hearing outcome is relatively good in a population not treated with routine insertion of ventilation tubes. The majority of patients also have satisfactory speech. Patients with cleft palate should have close follow-up for middle ear disease, but further research is warranted to determine the aggressive usage of ventilation tubes. PMID- 12733962 TI - Characteristics of nonverbal behavior in patients with cleft lip and palate during interpersonal communication. AB - OBJECTIVE: This study examined characteristics of nonverbal behavior that patients with cleft lip and palate (CLP) presented during interpersonal communication. DESIGN: This was a case-control design comparing nonverbal behavior of adult women with CLP with females without CLP. PARTICIPANTS: Subjects were 20 adult women with CLP and 20 noncleft control women matched for age and educational experience. MAIN OUTCOME MEASURES: Subject gestures and facial expressions were videotaped during interviews and analyzed with a computer-based kinematic measurement system. RESULTS: The clinical group displayed significantly fewer head movements and a lower smile frequency than the control group. Furthermore, head and hand movements and smiles were less coordinated or congruent for the subjects with CLP than for the comparison group. CONCLUSIONS: Even slight facial disfigurement could have a harmful effect on communication behavior in female patients with CLP. PMID- 12733963 TI - Unilateral advancement of the maxillary minor segment by distraction osteogenesis in patients with repaired unilateral cleft lip and palate: report of two cases. AB - OBJECTIVE: Collapse of the maxillary minor segment with lateral crossbite is a common feature in patients with repaired unilateral cleft lip/palate because of maxillary alveolar bony defect and palatal scar tissue. Distraction osteogenesis (DOG) is an effective technique of lengthening and augmentation for bone and gingiva. This case report describes the effects of unilateral advancement of the maxillary minor segment by DOG in two patients with the repaired unilateral cleft lip/palate. PMID- 12733964 TI - Pai syndrome: an adult patient with bifid nose and frontal hairline marker. AB - OBJECTIVE: A 17-year-old previously unreported patient with Pai syndrome is described. The boy had median cleft of upper lip, a polypoid skin mass over the columella, a minimal cleft of the upper central incisors, frontal alopecia of the anterior hairline, and bifid nose. Magnetic resonance imaging showed pericallosal lipoma. No mental retardation was present, and a chromosomal study showed normal male 46, XY karyotype. PMID- 12733966 TI - Application of a new method of nonlinear dynamical system identification to biochemical problems. AB - The system identification method for a variety of nonlinear dynamic models is elaborated. The problem of identification of an original nonlinear model presented as a system of ordinary differential equations in the Cauchy explicit form with a polynomial right part reduces to the solution of the system of linear equations for the constants of the dynamical model. In other words, to construct an integral model of the complex system (phenomenon), it is enough to collect some data array characterizing the time-course of dynamical parameters of the system. Collection of such a data array has always been a problem. However difficulties emerging are, as a rule, not principal and may be overcome almost without exception. The potentialities of the method under discussion are demonstrated by the example of the test problem of multiparametric nonlinear oscillator identification. The identification method proposed may be applied to the study of different biological systems and in particular the enzyme kinetics of complex biochemical reactions. PMID- 12733965 TI - Sphenoethmoidal encephalocele: a case report. AB - OBJECTIVE: This article documents the characteristics and treatment of an infant patient with a sphenoethmoidal encephalocele. An extracranial, transpalatal approach was used to eliminate the encephalocele. PMID- 12733967 TI - Soluble tankyrase located in cytosol of human embryonic kidney cell line 293. AB - We studied the subcellular localization of tankyrase in primary and immortalized human cell cultures. In embryonic kidney cell line 293 the enzyme was excluded from the nuclei and distributed in fractions of soluble cytosolic proteins and low-density microsomes. Newly revealed cytosolic tankyrase in its poly(ADP ribosyl)ated form was passed through a Sepharose 2B column and eluted as an apparently monomeric protein. The cytosolic localization of the enzyme correlated with its relatively high activity in the 293 cell line in comparison to eight other studied cell types. PMID- 12733968 TI - Alpha-crystallin promotes assembly of a trimeric form of Mycobacterium tuberculosis Hsp16.3 in a cell free system. AB - Hsp16.3, a small heat shock protein from Mycobacterium tuberculosis proposed to form specific trimer-of-trimers structures, acts as a molecular chaperone in vitro. The assembly mechanisms of this oligomeric protein were studied using in vitro transcription/translation systems. Analysis using a combination of non denaturing pore gradient polyacrylamide gel electrophoresis and size-exclusion chromatography demonstrates that the predominant form of Hsp16.3 produced in the in vitro transcription/translation system is the trimer. Our result indicated that alpha-crystallin (molecular chaperone) remarkably promotes the trimer assembly of Hsp16.3, but does not convert the trimeric form to nonameric form. An "inert" Hsp16.3 dimer, which does not seem to participate in trimer assembly but may be involved in forming other forms of Hsp16.3, was also detected in the in vitro expression system. A latent phase of ~10 min in the appearance of the first detectable species indicated that Hsp16.3 assembly did not occur co translationally. PMID- 12733969 TI - A new antigenic epitope appears in the catalytic subunit of viscumin during intracellular transport. AB - The plant toxin viscumin (60 kD) consists of B- ("binding") and A- ("active") subunits joined by a disulfide bond. The B-subunit is a lectin interacting with galactose-containing glycolipids and glycoproteins of the cell surface. The A subunit possesses N-glycosidase activity which modifies 28S ribosomal RNA. This results in irreversible inhibition of protein synthesis. After binding and receptor-mediated endocytosis viscumin-containing vesicles are transported to endoplasmic reticulum where the A- (catalytic) subunit is subsequently translocated to cytosol. It is possible that translocation of A-subunit requires its unfolding. For identification of epitopes which might appear during such unfolding, we developed hybridomas producing monoclonal antibodies against denatured viscumin A-chain. Resistance of hybridoma cells to cytotoxic action of viscumin suggests antibody-toxin interaction inside these cells. TA7 hybridoma cells against an epitope which appears only in denatured viscumin are insensitive to the toxin. This suggests that antibody-toxin interaction occurs before transmembrane translocation of the catalytic A-chain into the cytoplasm. Consequently, toxin resistance of TA7 hybridoma cells implies the appearance of a new epitope in viscumin during its intracellular transportation inside of vesicles. Sixty five octapeptides have been synthesized and epitopes have been identified for monoclonal TA7 antibody and immune mouse serum by means of ELISA. Based on the epitopic mapping the peptide A96-ETHLFTGT-T105 was chemically synthesized and binding of this peptide to the monoclonal antibody TA7 and conformation of antigenic determinant (L100-FTGT-T105) was investigated by means of (1)H-NMR spectroscopy. PMID- 12733970 TI - Molecular mechanisms of tumor angiogenesis. AB - The maintenance of growth of malignant tumors is closely related with the development of the vascular network supplying the tumor with blood. The vascularization of tumor tissue is similar to physiological angiogenesis, but in tumors it has some specific features. During the last 25 years a vast number of biomolecules have been found and described which are involved in the regulation of tumor angiogenesis. This review considers the action mechanisms and specific features of expression of the main angiogenic growth factors, such as the vascular endothelium growth factor (VEGF), angiopoietins (Ang-1, Ang-2), and the basic fibroblast growth factor (bFGF). The roles of cytokines, growth factors, proteolytic enzymes, and cell adhesion molecules in the regulation of the key steps of blood vessel generation in the tumor are considered. The significance of angiogenesis in the treatment of oncological diseases and possible approaches for inhibition of the regulatory signals of angiogenic factors are discussed. PMID- 12733971 TI - Thermostable DNA-polymerase from the thermophilic archaeon microorganism Archaeoglobus fulgidus VC16 and its features. AB - A gene (No. AF0497 GenBank, USA) was cloned from the archaeon Archaeoglobus fulgidus strain found in the water of hot springs. This gene contains an open reading frame of 2346 base pairs which encodes a thermostable DNA-polymerase (762 amino acid residues). A recombinant protein Afu-pol with molecular weight of 94 kD was isolated in an Escherichia coli strain used as a producer and characterized. By site-directed mutagenesis in the afu-pol gene the amino acid residue Glu170 was replaced with Ala; this resulted in a complete loss of the 3; 5;-exonuclease activity of the enzyme. Thus, the Glu170 residue was suggested to be directly involved in formation of the 3;-5;-exonuclease site. Physicochemical features of the exodeficient enzyme form were studied, and the possible use of Afu(exo(-))-pol in the polymerase chain reaction is shown. PMID- 12733972 TI - Graspases--a special group of serine proteases of the chymotrypsin family that has lost a conserved active site disulfide bond. AB - In this report we propose a new approach to classification of serine proteases of the chymotrypsin family. Comparative structure-function analysis has revealed two main groups of proteases: a group of trypsin-like enzymes and graspases (granule associated proteases). The most important structural peculiarity of graspases is the absence of conservative "active site" disulfide bond Cys191-Cys220. The residue at position 226 in the S1-subsite of graspases is responsible for substrate specificity, whereas the residue crucial for specificity in classical serine proteases is located at position 189. We distinguish three types of graspases on the base of their substrate specificity: 1) chymozymes prefer uncharged substrates and contain an uncharged residue at position 226; 2) duozymes possess dual trypsin-like and chymotrypsin-like specificity and contain Asp or Glu at 226; 3) aspartases hydrolyze Asp-containing substrates and contain Arg residue at 226. The correctness of the proposed classification was confirmed by phylogenic analysis. PMID- 12733973 TI - Distribution of O-glycosylhydrolases in marine invertebrates. Enzymes of the marine mollusk Littorina kurila that catalyze fucoidan transformation. AB - The distribution of O-glycosylhydrolases (fucoidan hydrolases, alpha-D mannosidases, beta-D-glucosidases, and beta-D-galactosidases) in 30 species of marine invertebrates occurring in the Sea of Japan was studied. It is shown that fucoidanases and glycosidases are widespread in the animals analyzed. Some molluscan, annelid, and echinoderm species can probably serve as objects for isolation and detailed study of the fucoidan-hydrolyzing enzymes. Fucoidan hydrolase, alpha-L-fucosidase, and arylsulfatase from the marine mollusk Littorina kurila were isolated and described. It was found that alpha-L fucosidase and arylsulfatase hydrolyze synthetic substrates and cannot hydrolyze natural fucoidan, whereas fucoidan hydrolase cleaves fucoidan to produce sulfated oligosaccharides and fucose. PMID- 12733974 TI - Levels of plasma vitamin E, vitamin C, TBARS, and cholesterol in male patients with colorectal tumors. AB - Vitamin E and vitamin C are involved in the defense of the body against free radical and reactive oxygen molecule induced damage. The best characterized biological damage caused by radicals is known as lipid peroxidation. Free radical formation is known to play a major role in the development of cancer. In this study, we measured plasma levels of thiobarbituric acid reactive substances (TBARS) as a marker of lipid peroxidation, cholesterol, and vitamins E and C as antioxidants in male patients with colorectal tumors (n = 20, 54.5 +/- 8.3 years). The patients had significantly higher plasma TBARS levels than age matched healthy subjects (p < 0.001). Plasma vitamin C levels were significantly lower in the patients compared to the healthy subjects (p < 0.001). On the other hand, plasma vitamin E levels in the patients were similar to those of healthy subjects. Plasma cholesterol levels were also found to be significantly elevated in patients with colorectal tumors (p < 0.001). Our results suggest that there is an imbalance between oxidant and antioxidant status in tumor genesis. PMID- 12733975 TI - Effects of the water-miscible organic solvents on lactoperoxidase purified from creek-water buffalo milk. AB - Water buffalo lactoperoxidase (WBLPO) was purified with Amberlite CG-50 (NH(4)(+) form) resin, CM-Sephadex C-50 ion-exchange chromatography, and Sephadex G-100 gel filtration chromatography from skimmed buffalo milk. The purity of the WBLPO was shown with SDS-PAGE. The R(z) (A(412)/A(280)) value for the WBLPO was 0.9. The optimum pH for the WBLPO was at 6.0. The K(m) value at optimum pH and 25 degrees C was 0.13 mM. The V(max) value at optimum pH and 25 degrees C was 5.3 micro mol/min per ml. The K(i) values for methanol, ethanol, dimethyl sulfoxide (DMSO), acetonitrile, isopropanol, tetrahydrofuran (THF), N,N-dimethylformamide (DMF), and ethylene glycol were 1.087, 0.364, 0.302, 0.459, 0.330, 0.126, 0.093, and 2.125 M, respectively. All the solvents showed competitive inhibition. The I(50) values of methanol, ethanol, dimethyl sulfoxide, acetonitrile, isopropanol, tetrahydrofuran, N,N-dimethylformamide, and ethylene glycol were 2.910, 0.942, 0.537, 1.320, 0.875, 0.470, 0.405, and 3.920 M, respectively. Ethylene glycol, methanol, acetonitrile, and ethanol have been found to be very promising solvents for performing biocatalytic reactions with LPO in organic media. PMID- 12733976 TI - Study of nucleophile binding in the penicillin acylase active center. Kinetic analysis. AB - The influence of the external nucleophile (6-aminopenicillanic acid) on the kinetics of the penicillin acylase-catalyzed acyl transfer reactions was studied using a highly sensitive spectrophotometric assay. An adequate kinetic scheme is suggested based on kinetic analysis of the experimental dependencies of the k(cat) and K(m) values on the nucleophile concentration. The proposed kinetic scheme has been verified by a quantitative description of the above-mentioned experimental dependencies using the set of kinetic parameters obtained from independent experiments. Such an approach can be used for modeling of different penicillin acylase-catalyzed acyl transfer reactions. PMID- 12733977 TI - Anti-plasminogen autoantibodies from plasma of patients with systemic lupus erythematosus having anti-phospholipid antibody syndrome: isolation and some immunochemical properties. AB - Blood plasma samples from patients with systemic lupus erythematosus having the anti-phospholipid antibody syndrome were found to contain anti-plasminogen antibodies of the IgG class. The titers of anti-plasminogen autoantibodies of the IgG class were elevated in these patients compared with normal controls. Part of the pool of IgG anti-plasminogen antibodies reacts with an epitope in the lysine binding sites of plasminogen. Anti-plasminogen IgG isolated from patients' blood plasma is specific only for a native epitope of human plasminogen passively adsorbed on immunosorbent micro-titration plate. As shown by enzyme immunoassay, autoantibodies to plasminogen of the IgG class cross-react with human fibrinogen. PMID- 12733978 TI - TBARS, carnitine, and reduced glutathione levels in human bladder carcinoma. AB - In this study, we investigated tissue levels of reduced glutathione (GSH) and carnitine as well as thiobarbituric acid reactive substances (TBARS, as a marker of lipid peroxidation) levels in bladder carcinoma and control group of patients. The average GSH, carnitine and TBARS levels for tumor group were respectively 7.11 +/- 3.3 micro g/mg protein, 1.81 +/- 0.39 nmol/mg protein, and 4.29 +/- 3.2 micro mol/mg protein, versus 14.45 +/- 4.11 micro g/mg protein, 2.14 +/- 0.66 nmol/mg protein, and 2.3 +/- 0.6 micro mol/mg protein for normal bladder tissues. Thus, tissue reduced glutathione levels (GSH) were significantly lower in patients as compared with the control group (p < 0.001) whereas average TBARS levels in the tumor group were found to be higher than those in control group. The average tissue carnitine levels in the patient group were found to be lower compared with the control group but the difference was not statistically significant (p > 0.05). PMID- 12733979 TI - Concentration oscillations in three-component reaction systems. AB - Based on the sufficient conditions for the existence of oscillation modes in the mathematical models of biological systems, simple three-component schemes of chemical reactions with oscillatory behavior including only mono- and bimolecular steps are suggested. PMID- 12733980 TI - Occupancy of two primary chloride-binding sites in Natronobacterium pharaonis halorhodopsin is a necessary condition for active anion transport. AB - The existence of two primary chloride-binding sites was found on the basis of the study of halorhodopsin spectra at different chloride concentrations. SVD analysis of the spectra revealed two chloride-dependent components at low chloride concentration (0.1-10 mM). Global fitting of SVD components found K(D) values of 0.47 and 5.2 mM with unitity Hill coefficients. The second K(D) coincides with the apparent K(D) of the photovoltage response of halorhodopsin. PMID- 12733981 TI - Protein farnesyltransferase inhibitors. AB - Specific mutations in the ras gene impair the guanosine triphophatase (GTPase) activity of Ras proteins, which play a fundamental role in the signaling cascade, leading to uninterrupted growth signals and to the transformation of normal cells into malignant phenotypes. It has been shown that normal cells transfected with mutant ras gene become cancerous and that unfarnesylated, cytosolic mutant Ras protein does not anchor onto cell membranes and cannot induce this transformation. Posttranslational modification and plasma membrane association of mutant Ras is necessary for this transforming activity. Since its identification, the enzyme protein farnesyltransferase (FTase) that catalyzes the first and essential step of the three Ras-processing steps has emerged as the most promising target for therapeutic intervention. FTase has been implicated as a potential target in inhibiting the prenylation of a variety of proteins, thus in controlling varied disease states (e.g. cancer, neurofibromatosis, restenosis, viral hepatitis, bone resorption, parasitic infections, corneal inflammations, and diabetes) associated with prenyl modifications of Ras and other proteins. Furthermore, it has been suggested that FTase inhibitors indirectly help in inhibiting tumors via suppression of angiogenesis and induction of apoptosis. Major milestones have been achieved with small-molecule FTase inhibitors that show efficacy without toxicity in vitro, as well as in mouse models bearing ras dependent tumors. With the determination of the crystal structure of mammalian FTase, existent leads have been fine-tuned and new potent molecules of diverse structural classes have been designed. A few of these molecules are currently in the clinic, with at least three drug candidates in Phase II studies and one in Phase III. This article will review the progress that has been reported with FTase inhibitors in drug discovery and in the clinic. PMID- 12733983 TI - Heterocyclic compounds as inflammation inhibitors. AB - Clinical use of non-steroidal anti-inflammatory drugs (NSAIDs) is associated with significant toxicity particularly in the gastrointestinal tract and kidney. Various approaches such as formulation co-administration (of agents to protect the stomach), chemical manipulation and synthesis of new safer anti-inflammatory drugs reported in the literature to overcome the toxicity of NSAIDs have been summarized. As far as synthesis of new more effective and safer anti-inflammatory drugs is concerned, we have reported recent findings in the area of synthesis of heterocyclic compounds such as pyrimidines, imidazole, benzimidazole, thiazole, thiazolidine, acridine, thiourea, alkanoic acid derivatives and other related heterocyclic compounds and their role as inflammation inhibitors. PMID- 12733982 TI - Selective COX-2 inhibitors and dual acting anti-inflammatory drugs: critical remarks. AB - Non steroidal anti-inflammatory drugs (NSAIDs) are still the most commonly used remedies for rheumatic diseases. But NSAIDs produce serious adverse effects, the most important being gastric injury up to gastric ulceration and renal damage. Several strategies have been adopted in order to avoid these shortcomings, especially gastrointestinal toxicity. So, non steroidal anti-inflammatory drugs have been associated with gastroprotective agents that counteract the damaging effects of prostaglandin synthesis suppression: however, a combination therapy introduces problems of pharmacokinetics, toxicity, and patient s compliance. Also incorporation of a nitric oxide (NO)-generating moiety into the molecule of several NSAIDs was shown to greatly attenuate their ulcerogenic activity: however, several findings suggest a possible involvement of NO in the pathogenesis of arthritis and subsequent tissue destruction. A most promising approach seemed to be the preparation of novel NSAIDs, specific for the inducible isoform of cyclooxygenase (COX-2): they appear to be devoid of gastrointestinal toxicity, in that they spare mucosal prostaglandin synthesis. However, a number of recent studies raised serious questions about the two central tenets that support this approach, namely that the prostaglandins that mediate inflammation and pain are produced solely via COX-2 and that the prostaglandins that are important in gastrointestinal and renal function are produced solely via COX-1. So, increasing evidence shows that COX-2 (not only COX-1) also plays a physiological role in several body functions and that, conversely, COX-1 (not only COX-2) may also be induced at sites of inflammation. Moreover, COX-2 selective NSAIDs have lost the cardiovascular protective effects of non-selective NSAIDs, effects which are mediated through COX-1 inhibition (in addition, COX-2 has a role in sustaining vascular prostacyclin production). The products generated by the 5-lipoxygenase pathway (leukotrienes) are particularly important in inflammation: indeed, leukotrienes increase microvascular permeability and are potent chemotactic agents; moreover, inhibition of 5-lipoxygenase indirectly reduces the expression of TNF-alpha (a cytokine that plays a key role in inflammation). This explains the efforts to obtain drugs able to inhibit both 5 lipoxygenase and cyclooxygenases: the so-called dual acting anti-inflammatory drugs. Such compounds retain the activity of classical NSAIDs, while avoiding their main drawbacks, in that curtailed production of gastroprotective prostaglandins is associated with a concurrent curtailed production of the gastro damaging and bronchoconstrictive leukotrienes. Moreover, thanks to their mechanism of action, dual acting anti-inflammatory drugs could not merely alleviate symptoms of rheumatic diseases, but might also satisfy, at least in part, the criteria of curative drugs. Indeed, leukotrienes are pro-inflammatory, increase microvascular permeability, are potent chemotactic agents and attract eosinophils, neutrophils and monocytes into the synovium. Finally, recent data strongly suggest that dual inhibitors may have specific protective activity also in neurodegeneration. PMID- 12733984 TI - Insect pheromone olfaction: new targets for the design of species-selective pest control agents. AB - Insects communicate extensively with chemical signals specific to their species (pheromones). The highly sensitive and selective olfactory system involved in detection of these signals has attracted attention as a target for the design of novel pest control agents. This review summarizes efforts in understanding the structure-activity relationships of pheromone olfaction and in the design of compounds that selectively interfere with transport, recognition and degradation of pheromones in the peripheral olfactory system. Pheromone olfaction inhibitors are potential environmentally benign insect control agents. PMID- 12733985 TI - Interaction of saposin D with membranes: effect of anionic phospholipids and sphingolipids. AB - Saposin (Sap) D is an endolysosomal protein that, together with three other similar proteins, Sap A, Sap B and Sap C, is involved in the degradation of sphingolipids and, possibly, in the solubilization and transport of gangliosides. We found that Sap D is able to destabilize and disrupt membranes containing each of the three anionic phospholipids most abundant in the acidic endolysosomal compartment, namely lysobisphosphatidic acid (LBPA), phosphatidylinositol (PI) and phosphatidylserine (PS). The breakdown of the membranes, which occurs when the Sap D concentration on the lipid surface reaches a critical value, is a slow process that gives rise to small particles. The Sap D-particle complexes formed in an acidic milieu can be dissociated by an increase in pH, suggesting a dynamic association of Sap D with membranes. The presence of anionic phospholipids is required also for the Sap D-induced perturbation and solubilization of membranes containing a neutral sphingolipid such as ceramide or a ganglioside such as G(M1). At appropriate Sap D concentrations Cer and G(M1) are solubilized as constituents of small phospholipid particles. Our findings imply that most functions of Sap D are dependent on its interaction with anionic phospholipids, which mediate the Sap D effect on other components of the membrane such as sphingolipids. On consideration of the properties of Sap D we propose that Sap D might have a role in the definition of the structure and function of membranes, such as the intra-endolysosomal membranes, that are rich in anionic phospholipids. PMID- 12733986 TI - Structure of xylose reductase bound to NAD+ and the basis for single and dual co substrate specificity in family 2 aldo-keto reductases. AB - The co-ordinates reported have been submitted to the Protein Data Bank under accession number 1MI3. Xylose reductase (XR; AKR2B5) is an unusual member of aldo keto reductase superfamily, because it is one of the few able to efficiently utilize both NADPH and NADH as co-substrates in converting xylose into xylitol. In order to better understand the basis for this dual specificity, we have determined the crystal structure of XR from the yeast Candida tenuis in complex with NAD(+) to 1.80 A resolution (where 1 A=0.1 nm) with a crystallographic R factor of 18.3%. A comparison of the NAD(+)- and the previously determined NADP(+)-bound forms of XR reveals that XR has the ability to change the conformation of two loops. To accommodate both the presence and absence of the 2' phosphate, the enzyme is able to adopt different conformations for several different side chains on these loops, including Asn(276), which makes alternative hydrogen-bonding interactions with the adenosine ribose. Also critical is the presence of Glu(227) on a short rigid helix, which makes hydrogen bonds to both the 2'- and 3'-hydroxy groups of the adenosine ribose. In addition to changes in hydrogen-bonding of the adenosine, the ribose unmistakably adopts a 3'- endo conformation rather than the 2'- endo conformation seen in the NADP(+)-bound form. These results underscore the importance of tight adenosine binding for efficient use of either NADH or NADPH as a co-substrate in aldo-keto reductases. The dual specificity found in XR is also an important consideration in designing a high-flux xylose metabolic pathway, which may be improved with an enzyme specific for NADH. PMID- 12733987 TI - Structural and thermal stability analysis of Escherichia coli and Alicyclobacillus acidocaldarius thioredoxin revealed a molten globule-like state in thermal denaturation pathway of the proteins: an infrared spectroscopic study. AB - The structure of thioredoxin from Alicyclobacillus acidocaldarius (previously named Bacillus acidocaldarius ) (BacTrx) and from Escherichia coli ( E. coli Trx) was studied by Fourier-transform IR spectroscopy. Two mutants of BacTrx [Lys(18)- >Gly (K18G) and Arg(82)-->Glu (R82E)] were also analysed. The data revealed similar secondary structures in all proteins, but BacTrx and its mutants showed a more compact structure than E. coli Trx. In BacTrx and its mutants, the compactness was p(2)H-dependent. All proteins revealed the existence of a molten globule-like state. At p(2)H 5.8, the temperature at which this state was detected was higher in BacTrx and decreased in the different proteins in the following order: BacTrx>R82E>K18G> E. coli Trx. At neutral or basic p(2)H, the molten globule-like state was detected at the same temperature in both BacTrx and R82E, whereas it was found at the same temperature in all p(2)Hs tested for E. coli Trx. The thermal stability of the proteins was in the following order at all p(2)Hs tested: BacTrx>R82E>K18G> E. coli Trx, and was lower for each protein at p(2)H 8.4 than at neutral or acidic p(2)Hs. The formation of protein aggregates, brought about by thermal denaturation, were observed for BacTrx and K18G at all p(2)Hs tested, whereas they were present in R82E and E. coli Trx samples only at p(2)H 5.8. The results indicated that a single mutation might affect the structural properties of a protein, including its propensity to aggregate at high temperatures. The data also indicated a possible application of Fourier-transform IR spectroscopy for assessing molten globule-like states in small proteins. PMID- 12733988 TI - Differential signalling by muscarinic receptors in smooth muscle: m2-mediated inactivation of myosin light chain kinase via Gi3, Cdc42/Rac1 and p21-activated kinase 1 pathway, and m3-mediated MLC20 (20 kDa regulatory light chain of myosin II) phosphorylation via Rho-associated kinase/myosin phosphatase targeting subunit 1 and protein kinase C/CPI-17 pathway. AB - Signalling via m3 and m2 receptors in smooth muscles involved activation of two G protein-dependent pathways by each receptor. m2 receptors were coupled via Gbetagammai3 with activation of phospholipase C-beta3, phosphoinositide 3-kinase and Cdc42/Rac1 (where Cdc stands for cell division cycle) and p21-activated kinase 1 (PAK1), resulting in phosphorylation and inactivation of myosin light chain kinase (MLCK). Each step was inhibited by methoctramine and pertussis toxin. PAK1 activity was abolished in cells expressing both Cdc42-DN (where DN stands for dominant negative) and Rac1-DN. MLCK phosphorylation was inhibited by PAK1 antibody, and in cells expressing Cdc42-DN and Rac1-DN. m3 receptors were coupled via Galpha(q/11) with activation of phospholipase C-beta1 and via RhoA with activation of Rho-associated kinase (Rho kinase), phospholipase D and protein kinase C (PKC). Rho kinase and phospholipase D activities were inhibited by C3 exoenzyme and in cells expressing RhoA-DN. PKC activity was inhibited by bisindolylmaleimide, and in cells expressing RhoA-DN; PKC activity was also inhibited partly by Y27632 (44+/-5%). PKC-induced phosphorylation of PKC activated 17 kDa inhibitor protein of type 1 phosphatase (CPI-17) at Thr38 was abolished by bisindolylmaleimide and inhibited partly by Y27632 (28+/-3%). Rho kinase-induced phosphorylation of myosin phosphatase targeting subunit (MYPT1) and was abolished by Y27632. Sustained phosphorylation of 20 kDa regulatory light chain of myosin II (MLC20) and contraction were abolished by bisindolylmaleimide Y27632 and C3 exoenzyme and in cells expressing RhoA-DN. The results suggest that Rho-kinase-dependent phosphorylation of MYPT1 and PKC-dependent phosphorylation and enhancement of CPI-17 binding to the catalytic subunit of MLC phosphatase (MLCP) act co-operatively to inhibit MLCP activity, leading to sustained stimulation of MLC20 phosphorylation and contraction. Because Y27632 inhibited both Rho kinase and PKC activities, it could not be used to ascertain the contribution of MYPT1 to inhibition of MLCP activity. m2-dependent phosphorylation and inactivation of MLCK precluded its involvement in sustained MLC20 phosphorylation and contraction. PMID- 12733989 TI - Comparative membrane interaction study of viscotoxins A3, A2 and B from mistletoe (Viscum album) and connections with their structures. AB - Viscotoxins A2 (VA2) and B (VB) are, together with viscotoxin A3 (VA3), among the most abundant viscotoxin isoforms that occur in mistletoe-derived medicines used in anti-cancer therapy. Although these isoforms have a high degree of amino-acid sequence similarity, they are strikingly different from each other in their in vitro cytotoxic potency towards tumour cells. First, as VA3 is the only viscotoxin whose three-dimensional (3D) structure has been solved to date, we report the NMR determination of the 3D structures of VA2 and VB. Secondly, to account for the in vitro cytotoxicity discrepancy, we carried out a comparative study of the interaction of the three viscotoxins with model membranes. Although the overall 3D structure is highly conserved among the three isoforms, some discrete structural features and associated surface properties readily account for the different affinity and perturbation of model membranes. VA3 and VA2 interact in a similar way, but the weaker hydrophobic character of VA2 is thought to be mainly responsible for the apparent different affinity towards membranes. VB is much less active than the other two viscotoxins and does not insert into model membranes. This could be related to the occurrence of a single residue (Arg25) protruding outside the hydrophobic plane formed by the two amphipathic alpha-helices, through which viscotoxins are supposed to interact with plasma membranes. PMID- 12733993 TI - A marriage of techniques. PMID- 12733990 TI - S3 to S3' subsite specificity of recombinant human cathepsin K and development of selective internally quenched fluorescent substrates. AB - We have systematically examined the S3 to S3' subsite substrate specificity requirements of cathepsin K using internally quenched fluorescent peptides derived from the lead sequence Abz-KLRFSKQ-EDDnp [where Abz is o -aminobenzoic acid and EDDnp is N -(2,4-dinitrophenyl)ethylenediamine]. We assayed six series of peptides, in which each position except Gln was substituted with various natural amino acids. The results indicated that the S3-S1 subsite requirements are more restricted than those of S1'-S3'. Cathepsin K preferentially accommodates hydrophobic amino acids with aliphatic side chains (Leu, Ile and Val) in the S2 site. Modifications at P1 residues also have a large influence on cathepsin K activity. Positively charged residues (Arg and Lys) represent the best accepted amino acids in this position, although a particular preference for Gly was found as well. Subsite S3 accepted preferentially basic amino acids such as Lys and Arg. A broad range of amino acids was accommodated in the remaining subsites. We further explored the acceptance of a Pro residue in the P2 position by cathepsin K in order to develop specific substrates for the enzyme. Two series of peptides with the general sequences Abz-KXPGSKQ-EDDnp and Abz-KPXGSKQ-EDDnp (where X denotes the position of the amino acid that is altered) were synthesized. The substrates Abz-KPRGSKQ-EDDnp and Abz-KKPGSKQ-EDDnp were cleaved by cathepsin K at the Arg-Gly and Gly-Ser bonds respectively, and have been shown to be specific for cathepsin K when compared with other lysosomal cysteine proteases such as cathepsins L and B and with the aspartyl protease cathepsin D. PMID- 12733991 TI - Characterization of the human activator protein-2gamma (AP-2gamma) gene: control of expression by Sp1/Sp3 in breast tumour cells. AB - The activator protein-2 (AP-2) family of DNA-binding transcription factors are developmentally regulated and also play a role in human neoplasia. In particular, the AP-2gamma protein has been shown to be overexpressed in a high percentage of breast tumours. In the present study, we report the complete sequence determination of the human TFAP2C gene encoding the AP-2gamma transcription factor plus the mapping of the transcription start site used in breast tumour derived cells. The 5'-end of the gene lies within a CpG island and transcription is initiated at a single site within a classical initiator motif. We have gone on to investigate why some breast tumour-derived cell lines readily express AP 2gamma, whereas others do not, and show that the proximal promoter (+191 to -312) is differentially active in the two cell phenotypes. DNase footprinting led to the identification of three Sp1/Sp3-binding sites within this region, two of which are absolutely required both for promoter function and cell-type-specific activity. By Western blotting a panel of expressing and non-expressing breast tumour lines we show that the latter have higher levels of Sp3. Furthermore, increasing Sp3 levels in AP-2gamma-expressing cells led to the repression of AP 2gamma promoter activity, particularly when Sp3 inhibitory function was maximized through sumoylation. We propose that differences in the level and activity of Sp3 between breast tumour lines can determine the expression level of their AP-2gamma gene. PMID- 12733994 TI - Growing without a size checkpoint. PMID- 12733995 TI - Combined single-molecule force and fluorescence measurements for biology. AB - Recent advances in single-molecule techniques allow the application of force to an individual biomolecule whilst simultaneously monitoring its response using fluorescent probes. The effects of applied mechanical load on single-enzyme turnovers, biomolecular interactions and conformational changes can now be studied with nanometer precision and millisecond time resolution. PMID- 12733996 TI - Controlling cell division in yeast and animals: does size matter? AB - In yeast, cell-size checkpoints coordinate cellular growth with cell-cycle progression. Now, evidence has been provided that such checkpoints probably do not exist in mammalian cells. These findings highlight an important difference between how yeast and animal cells proliferate in response to extracellular cues. PMID- 12733997 TI - Combined optical trapping and single-molecule fluorescence. AB - BACKGROUND: Two of the mainstay techniques in single-molecule research are optical trapping and single-molecule fluorescence. Previous attempts to combine these techniques in a single experiment - and on a single macromolecule of interest - have met with little success, because the light intensity within an optical trap is more than ten orders of magnitude greater than the light emitted by a single fluorophore. Instead, the two techniques have been employed sequentially, or spatially separated by distances of several micrometers within the sample, imposing experimental restrictions that limit the utility of the combined method. Here, we report the development of an instrument capable of true, simultaneous, spatially coincident optical trapping and single-molecule fluorescence. RESULTS: We demonstrate the capability of the apparatus by studying force-induced strand separation of a rhodamine-labeled, 15 base-pair segment of double-stranded DNA, with force applied perpendicular to the axis of the DNA molecule. As expected, we observed abrupt mechanical transitions corresponding to the unzipping of DNA at a critical force. Transitions occurred concomitant with changes in the fluorescence of dyes attached at the duplex ends, which became unquenched upon strand separation. CONCLUSIONS: Through careful optical design, the use of high-performance spectral notch filters, a judicious choice of fluorophores, and the rapid acquisition of data gained by computer-automating the experiment, it is possible to perform combined optical trapping and single molecule fluorescence. This opens the door to many types of experiment that employ optical traps to supply controlled external loads while fluorescent molecules report concurrent information about macromolecular structure. PMID- 12733999 TI - War and peace. AB - If the fiftieth anniversary of the double helix teaches us anything it ought to be that great scientific discoveries are almost never made in time of war or directly for the purpose of war. PMID- 12733998 TI - Differences in the way a mammalian cell and yeast cells coordinate cell growth and cell-cycle progression. AB - BACKGROUND: It is widely believed that cell-size checkpoints help to coordinate cell growth and cell-cycle progression, so that proliferating eukaryotic cells maintain their size. There is strong evidence for such size checkpoints in yeasts, which maintain a constant cell-size distribution as they proliferate, even though large yeast cells grow faster than small yeast cells. Moreover, when yeast cells are shifted to better or worse nutrient conditions, they alter their size threshold within one cell cycle. Populations of mammalian cells can also maintain a constant size distribution as they proliferate, but it is not known whether this depends on cell-size checkpoints. RESULTS: We show that proliferating rat Schwann cells do not require a cell-size checkpoint to maintain a constant cell-size distribution, as, unlike yeasts, large and small Schwann cells grow at the same rate, which depends on the concentration of extracellular growth factors. In addition, when shifted from serum-free to serum-containing medium, Schwann cells take many divisions to increase their size to that appropriate to the new condition, suggesting that they do not have cell-size checkpoints similar to those in yeasts. CONCLUSIONS: Proliferating Schwann cells and yeast cells seem to use different mechanisms to coordinate their growth with cell-cycle progression. Whereas yeast cells use cell-size checkpoints, Schwann cells apparently do not. It seems likely that many mammalian cells resemble Schwann cells in this respect. PMID- 12734000 TI - Genomic analysis of the eukaryotic protein kinase superfamily: a perspective. AB - Protein kinases with a conserved catalytic domain make up one of the largest 'superfamilies' of eukaryotic proteins and play many key roles in biology and disease. Efforts to identify and classify all the members of the eukaryotic protein kinase superfamily have recently culminated in the mining of essentially complete human genome data. PMID- 12734001 TI - Discovering human history from stomach bacteria. AB - Recent analyses of human pathogens have revealed that their evolutionary histories are congruent with the hypothesized pattern of ancient and modern human population migrations. Phylogenetic trees of strains of the bacterium Helicobacter pylori and the polyoma JC virus taken from geographically diverse groups of human beings correlate closely with relationships of the populations in which they are found. PMID- 12734002 TI - The role of DNA sequence in centromere formation. AB - Centromeres are key to the correct segregation and inheritance of genetic information. Eukaryotic centromeres, which are located in large blocks of highly repetitive DNA, have been notoriously difficult to sequence. Several groups have recently succeeded in analyzing centromeric sequences in human, Drosophila and Arabidopsis, providing new insights into the importance of DNA sequence for centromere function. PMID- 12734004 TI - The modular era of functional genomics. AB - A report on the Keystone Symposium 'Functional Genomics: Global Analysis of Complex Biological Systems', Santa Fe, USA, 20-24 February 2003. PMID- 12734003 TI - Expression profiling of plant development. AB - Microarrays have been used to study the response of plants to many signals, including light, hormones and transcription factors. The results in each case can give an overall view of the global response to the signal or identify direct targets of the signal, and can reveal new links between different signaling pathways. PMID- 12734006 TI - How is the Drosophila research community making use of the genome sequence? AB - A report on the 44th Annual Drosophila Research Conference, Chicago, USA, 5-9 March, 2003. PMID- 12734005 TI - Cancer and programmed cell death. AB - A report on the 15th Lorne Cancer Conference, Lorne, Australia, 13-16 February 2003. PMID- 12734007 TI - Unscrambling the genome. AB - A report on the 24th Annual Lorne Conference on the Organization and Expression of the Genome, Lorne, Victoria, Australia, 16-20 February 2003. PMID- 12734008 TI - Beyond 100 genomes. AB - By the end of 2002, we witnessed the landmark submission of the 100th complete genome sequence in the databases. An overview of these genomes reveals certain interesting trends and provides valuable insights into possible future developments. PMID- 12734009 TI - DAVID: Database for Annotation, Visualization, and Integrated Discovery. AB - BACKGROUND: Functional annotation of differentially expressed genes is a necessary and critical step in the analysis of microarray data. The distributed nature of biological knowledge frequently requires researchers to navigate through numerous web-accessible databases gathering information one gene at a time. A more judicious approach is to provide query-based access to an integrated database that disseminates biologically rich information across large datasets and displays graphic summaries of functional information. RESULTS: Database for Annotation, Visualization, and Integrated Discovery (DAVID; http://www.david.niaid.nih.gov) addresses this need via four web-based analysis modules: 1) Annotation Tool - rapidly appends descriptive data from several public databases to lists of genes; 2) GoCharts - assigns genes to Gene Ontology functional categories based on user selected classifications and term specificity level; 3) KeggCharts - assigns genes to KEGG metabolic processes and enables users to view genes in the context of biochemical pathway maps; and 4) DomainCharts - groups genes according to PFAM conserved protein domains. CONCLUSIONS: Analysis results and graphical displays remain dynamically linked to primary data and external data repositories, thereby furnishing in-depth as well as broad-based data coverage. The functionality provided by DAVID accelerates the analysis of genome-scale datasets by facilitating the transition from data collection to biological meaning. PMID- 12734010 TI - Identifying related L1 retrotransposons by analyzing 3' transduced sequences. AB - BACKGROUND: A large fraction of the human genome is attributable to L1 retrotransposon sequences. Not only do L1s themselves make up a significant portion of the genome, but L1-encoded proteins are thought to be responsible for the transposition of other repetitive elements and processed pseudogenes. In addition, L1s can mobilize non-L1, 3'-flanking DNA in a process called 3' transduction. Using computational methods, we collected DNA sequences from the human genome for which we have high confidence of their mobilization through L1 mediated 3' transduction. RESULTS: The precursors of L1s with transduced sequence can often be identified, allowing us to reconstruct L1 element families in which a single parent L1 element begot many progeny L1s. Of the L1s exhibiting a sequence structure consistent with 3' transduction (L1 with transduction-derived sequence, L1-TD), the vast majority were located in duplicated regions of the genome and thus did not necessarily represent unique insertion events. Of the remaining L1-TDs, some lack a clear polyadenylation signal, but the alignment between the parent-progeny sequences nevertheless ends in an A-rich tract of DNA. CONCLUSIONS: Sequence data suggest that during the integration into the genome of RNA representing an L1-TD, reverse transcription may be primed internally at A rich sequences that lie downstream of the L1 3' untranslated region. The occurrence of L1-mediated transduction in the human genome may be less frequent than previously thought, and an accurate estimate is confounded by the frequent occurrence of segmental genomic duplications. PMID- 12734011 TI - Duplication and selection in the evolution of primate beta-defensin genes. AB - BACKGROUND: Innate immunity is the first line of defense against microorganisms in vertebrates and acts by providing an initial barrier to microorganisms and triggering adaptive immune responses. Peptides such as beta-defensins are an important component of this defense, providing a broad spectrum of antimicrobial activity against bacteria, fungi, mycobacteria and several enveloped viruses. Beta-defensins are small cationic peptides that vary in their expression patterns and spectrum of pathogen specificity. Disruptions in beta-defensin function have been implicated in human diseases, including cystic fibrosis, and a fuller understanding of the variety, function and evolution of human beta-defensins might form the basis for novel therapies. Here we use a combination of laboratory and computational techniques to characterize the main human beta-defensin locus on chromosome 8p22-p23. RESULTS: In addition to known genes in the region we report the genomic structures and expression patterns of four novel human beta defensin genes and a related pseudogene. These genes show an unusual pattern of evolution, with rapid divergence between second exon sequences that encode the mature beta-defensin peptides matched by relative stasis in first exons that encode signal peptides. CONCLUSIONS: We conclude that the 8p22-p23 locus has evolved by successive rounds of duplication followed by substantial divergence involving positive selection, to produce a diverse cluster of paralogous genes established before the human-baboon divergence more than 23 million years ago. Positive selection, disproportionately favoring alterations in the charge of amino-acid residues, is implicated as driving second exon divergence in these genes. PMID- 12734014 TI - Clustering gene-expression data with repeated measurements. AB - Clustering is a common methodology for the analysis of array data, and many research laboratories are generating array data with repeated measurements. We evaluated several clustering algorithms that incorporate repeated measurements, and show that algorithms that take advantage of repeated measurements yield more accurate and more stable clusters. In particular, we show that the infinite mixture model-based approach with a built-in error model produces superior results. PMID- 12734012 TI - Combined histomorphometric and gene-expression profiling applied to toxicology. AB - We have developed a unique methodology for the combined analysis of histomorphometric and gene-expression profiles amenable to intensive data mining and multisample comparison for a comprehensive approach to toxicology. This hybrid technology, termed extensible morphometric relational gene-expression analysis (EMeRGE), is applied in a toxicological study of time-varied vehicle- and carbon-tetrachloride (CCl4)-treated rats, and demonstrates correlations between specific genes and tissue structures that can augment interpretation of biological observations and diagnosis. PMID- 12734013 TI - A novel exploratory method for visual recombination detection. AB - A versatile visual approach for detecting recombination and identifying recombination breakpoints within a sequence alignment is presented. The method is based on two novel diagrams - the highway plot and the occupancy plot - that graphically portray phylogenetic inhomogeneity along an alignment, and can be viewed as a synthesis of two widely used but unrelated methods: bootscanning and quartet-mapping. To illustrate the method, simulated data and HIV-1 and influenza A datasets are investigated. PMID- 12734015 TI - GeneHopper: a web-based search engine to link gene-expression platforms through GenBank accession numbers. AB - Global gene-expression analysis is carried out using different technologies that are either array- or sequence-tag-based. To compare experiments that are performed on these different platforms, array probes and sequence tags need to be linked. An additional challenge is cross-referencing between species, to compare human profiles with those obtained in a mouse model, for example. We have developed the web-based search engine GeneHopper to link different expression resources based on UniGene clusters and HomoloGene orthologs databases of the National Center for Biotechnology Information (NCBI). PMID- 12734016 TI - Content validity of manual spinal palpatory exams - A systematic review. AB - BACKGROUND: Many health care professionals use spinal palpatory exams as a primary and well-accepted part of the evaluation of spinal pathology. However, few studies have explored the validity of spinal palpatory exams. To evaluate the status of the current scientific evidence, we conducted a systematic review to assess the content validity of spinal palpatory tests used to identify spinal neuro-musculoskeletal dysfunction. METHODS: Review of eleven databases and a hand search of peer-reviewed literature, published between 1965-2002, was undertaken. Two blinded reviewers abstracted pertinent data from the retrieved papers, using a specially developed quality-scoring instrument. Five papers met the inclusion/exclusion criteria. RESULTS: Three of the five papers included in the review explored the content validity of motion tests. Two of these papers focused on identifying the level of fixation (decreased mobility) and one focused on range of motion. All three studies used a mechanical model as a reference standard. Two of the five papers included in the review explored the validity of pain assessment using the visual analogue scale or the subjects' own report as reference standards. Overall the sensitivity of studies looking at range of motion tests and pain varied greatly. Poor sensitivity was reported for range of motion studies regardless of the examiner's experience. A slightly better sensitivity (82%) was reported in one study that examined cervical pain. CONCLUSIONS: The lack of acceptable reference standards may have contributed to the weak sensitivity findings. Given the importance of spinal palpatory tests as part of the spinal evaluation and treatment plan, effort is required by all involved disciplines to create well-designed and implemented studies in this area. PMID- 12734017 TI - Structure determination of human Lck unique and SH3 domains by nuclear magnetic resonance spectroscopy. AB - BACKGROUND: Protein tyrosine kinases are involved in signal transduction pathways that regulate cell growth, differentiation, activation and transformation. Human lymphocyte specific kinase (Lck) is a 56 kDa protein involved in T-cell- and IL2 receptor signaling. Three-dimensional structures are known for SH3, SH2 and kinase domains of Lck as well as for other tyrosine kinases. No structure is known for the unique domain of any Src-type tyrosine kinase. RESULTS: Lck(1-120) comprising unique and SH3 domains was structurally investigated by nuclear magnetic resonance spectroscopy. We found the unique domain, in contrast to the SH3 part, to have basically no defined structural elements. The solution structure of the SH3 part could be determined with very high precision. It does not show significant differences to Lck SH3 in the absence of the unique domain. Minor differences were observed to the X-ray structure of Lck SH3. CONCLUSION: The unique domain of Lck does not contain any defined structure elements in the absence of ligands and membranes. Presence of the unique domain is not relevant to the three-dimensional structure of the Lck SH3 domain. PMID- 12734018 TI - Diagnostic properties of nerve conduction tests in population-based carpal tunnel syndrome. AB - BACKGROUND: Numerous nerve conduction tests are used for the electrodiagnosis of carpal tunnel syndrome (CTS), with a wide range of sensitivity and specificity reported for each test in clinical studies. The tests have not been assessed in population-based studies. Such information would be important when using electrodiagnosis in epidemiologic research. The purpose of this study was to compare the diagnostic accuracy of various nerve conduction tests in population based CTS and determine the properties of the most accurate test. METHODS: In a population-based study a questionnaire was mailed to a random sample of 3,000 persons. Of 2,466 responders, 262 symptomatic (numbness/tingling in the radial fingers) and 125 randomly selected asymptomatic responders underwent clinical and electrophysiologic examinations. A standardized hand diagram was administered to the symptomatic persons. At the clinical examination, the examining surgeon identified 94 symptomatic persons as having clinically certain CTS. Nerve conduction tests were then performed on the symptomatic and the asymptomatic persons by blinded examiners. Analysis with receiver operating characteristic (ROC) curves was used to compare the diagnostic accuracy of the nerve conduction tests in distinguishing the persons with clinically certain CTS from the asymptomatic persons. RESULTS: No difference was shown in the diagnostic accuracy of median nerve distal motor latency, digit-wrist sensory latency, wrist-palm sensory conduction velocity, and wrist-palm/forearm sensory conduction velocity ratio (area under curve, 0.75-0.76). Median-ulnar digit-wrist sensory latency difference had a significantly higher diagnostic accuracy (area under curve, 0.80). Using the optimal cutoff value of 0.8 ms for abnormal sensory latency difference shown on the ROC curve the sensitivity was 70%, specificity 82%, positive predictive value 19% and negative predictive value 98%. Based on the clinical diagnosis among the symptomatic persons, the hand diagram (classified as classic/probable or possible/unlikely CTS) had high sensitivity but poor specificity. CONCLUSIONS: Using the clinical diagnosis of CTS as the criterion standard, nerve conduction tests had moderate sensitivity and specificity and a low positive predictive value in population-based CTS. Measurement of median ulnar sensory latency difference had the highest diagnostic accuracy. The performance of nerve conduction tests in population-based CTS does not necessarily apply to their performance in clinical settings. PMID- 12734019 TI - A simple method for analyzing data from a randomized trial with a missing binary outcome. AB - BACKGROUND: Many randomized trials involve missing binary outcomes. Although many previous adjustments for missing binary outcomes have been proposed, none of these makes explicit use of randomization to bound the bias when the data are not missing at random. METHODS: We propose a novel approach that uses the randomization distribution to compute the anticipated maximum bias when missing at random does not hold due to an unobserved binary covariate (implying that missingness depends on outcome and treatment group). The anticipated maximum bias equals the product of two factors: (a) the anticipated maximum bias if there were complete confounding of the unobserved covariate with treatment group among subjects with an observed outcome and (b) an upper bound factor that depends only on the fraction missing in each randomization group. If less than 15% of subjects are missing in each group, the upper bound factor is less than.18. RESULTS: We illustrated the methodology using data from the Polyp Prevention Trial. We anticipated a maximum bias under complete confounding of.25. With only 7% and 9% missing in each arm, the upper bound factor, after adjusting for age and sex, was.10. The anticipated maximum bias of.25 x.10 =.025 would not have affected the conclusion of no treatment effect. CONCLUSION: This approach is easy to implement and is particularly informative when less than 15% of subjects are missing in each arm. PMID- 12734021 TI - An update on fluorides and fluorosis. AB - Decisions concerning use of fluoride in its many forms for caries prevention are more complicated now than in the past because of the need to balance these benefits with the risks of dental fluorosis. This article reviews pertinent literature concerning dental fluorosis (definition, appearance, prevalence), pre- and post-eruptive use of fluoride, esthetic perceptions of dental fluorosis, fluoride levels of beverages and foods, the Iowa Fluoride Study, and the U.S. Centers for Disease Control and Prevention's "Recommendations for Using Fluoride to Prevent and Control Dental Caries in the United States." Water fluoridation and use of fluoride dentifrice are the most efficient and cost-effective ways to prevent dental caries; other modalities should be targeted toward high-risk individuals. PMID- 12734020 TI - Domain fusion analysis by applying relational algebra to protein sequence and domain databases. AB - BACKGROUND: Domain fusion analysis is a useful method to predict functionally linked proteins that may be involved in direct protein-protein interactions or in the same metabolic or signaling pathway. As separate domain databases like BLOCKS, PROSITE, Pfam, SMART, PRINTS-S, ProDom, TIGRFAMs, and amalgamated domain databases like InterPro continue to grow in size and quality, a computational method to perform domain fusion analysis that leverages on these efforts will become increasingly powerful. RESULTS: This paper proposes a computational method employing relational algebra to find domain fusions in protein sequence databases. The feasibility of this method was illustrated on the SWISS PROT+TrEMBL sequence database using domain predictions from the Pfam HMM (hidden Markov model) database. We identified 235 and 189 putative functionally linked protein partners in H. sapiens and S. cerevisiae, respectively. From scientific literature, we were able to confirm many of these functional linkages, while the remainder offer testable experimental hypothesis. Results can be viewed at http://calcium.uhnres.utoronto.ca/pi. CONCLUSION: As the analysis can be computed quickly on any relational database that supports standard SQL (structured query language), it can be dynamically updated along with the sequence and domain databases, thereby improving the quality of predictions over time. PMID- 12734022 TI - Oral health promotion for high-risk children: case studies from British Columbia. AB - Socio-economics, family stress and parenting style each plays as important a role in the development of early childhood caries as dietary and microbiological factors. Therefore, to be successful, oral health promotion initiatives should be designed and implemented with due consideration of issues such as collaboration with community partners, the role of dental health in overall child health and the involvement of the community in program planning. This article briefly describes 3 programs that have been undertaken in British Columbia either as public health initiatives or as demonstration research projects to improve oral health in young children from diverse communities with a high prevalence of early childhood caries. PMID- 12734023 TI - Intravenous procedural sedation: an alternative to general anesthesia in the treatment of early childhood caries. AB - Providing comprehensive dental treatment for preschool children with early childhood caries (ECC) is probably the greatest challenge facing most dentists; many elect to hospitalize a child with ECC and provide treatment under general anesthesia. However, as waiting lists for hospital admission are long, ECC continues to progress and can cause pain or acute infection. Hospital treatment also results in substantial costs beyond those for dental treatment. This paper describes a cost-effective intravenous sedation program being used in a pediatric dental practice in Kelowna, British Columbia. The program offers an alternative to general anesthesia that allows qualified dentists to provide comprehensive dental treatment to children with ECC safely and efficiently in the private practice setting. PMID- 12734024 TI - Causes, treatment and prevention of early childhood caries: a microbiologic perspective. AB - Early childhood caries (ECC) is a virulent form of dental caries that can destroy the primary dentition of toddlers and preschool children. It occurs worldwide, afflicting predominantly disadvantaged children. High-risk North American populations include Hispanic and Native American children, as well as children enrolled in Head Start, a federally funded program for preschool children living in poverty. The prevalence of EEC among these children ranges from 11% to 72%. ECC is an infectious disease, and Streptococcus mutans is the most likely causative agent; diet also plays a critical role in the acquisition and clinical expression of this infection. Early acquisition of S. mutans is a key event in the natural history of the disease. Acquisition may occur via vertical or horizontal transmission. Primary oral colonization by S. mutans coupled with caries-promoting feeding behaviours results in accumulation of these organisms to levels exceeding 30% of the total cultivable plaque flora which in turn leads to rapid demineralization of tooth structure. Treatment of ECC is costly because the cooperative capacity of babies and preschool children usually necessitates the use of general anesthesia. Treatment usually consists of restoration or surgical removal of carious teeth along with recommendations regarding feeding habits. However, this approach has resulted in unacceptable clinical outcomes, and relapse rates of approximately 40% have been reported within the first year after dental surgery. Primary prevention of ECC has largely been restricted to counselling parents about caries-promoting feeding behaviours. This approach has also had minimal success. Newer strategies addressing the infectious component through use of topical antimicrobial therapy appear promising. PMID- 12734025 TI - Avulsions and intrusions: the controversial displacement injuries. AB - Avulsions and intrusions are the most complicated and controversial displacement injuries of permanent teeth. Clinical guidelines published by authorities such as the American Association of Endodontists, the Royal College of Surgeons of England and the International Association of Dental Traumatology are inconsistent. While a certain amount of inconsistency might be expected, some of these guidelines recommend treatments that are experimental or have not incorporated research information from the past 5 years, and in one case the guidelines incorrectly describe the nature of Hank's balanced salt solution. Recent laboratory studies support previous clinical outcome studies in emphasizing that only for teeth replanted within 5 minutes of avulsion is there a chance of regeneration of the periodontal ligament and normal function. Teeth replanted beyond 5 minutes will take another path, that of repair followed by root resorption, ankylosis and eventual extraction. Dentists should explain these outcomes at the time of the replantation decision. Severe intrusions also have predictable outcomes. Teeth intruded beyond 6 mm cannot regenerate a functional periodontal ligament and so are prone to root resorption and eventual extraction as well. In this situation the decision is one of immediate extraction or repositioning, with the understanding that it is inevitable that the tooth will eventually be extracted. Authoritative clinical guidelines available on the Internet provide the clinician with useful outlines for treatment. However, individual inconsistencies stimulate academic controversies and, in some cases, clinical misdirection. PMID- 12734026 TI - Domestic terrorism targets the medical profession. PMID- 12734027 TI - Long-term estrogen and hormone replacement therapy for the prevention and treatment of osteoporosis. AB - Recent studies have called into question whether the risks of hormone replacement therapy (HRT) outweigh its long-term benefits. There is a clear causal relationship between estrogen deficiency and osteoporosis. Postmenopausal status or estrogen-deficiency at any age significantly increases a patient's risk for osteoporosis and subsequent fragility fractures. Estrogen, in various formulations, is currently FDA-indicated for the prevention of osteoporosis. However, estrogen is not FDA-approved for the treatment of osteoporosis. Recent randomized clinical trials have demonstrated that estrogen significantly reduces fractures among osteoporotic postmenopausal women. Most postmenopausal women choose to use estrogen for relief of vasomotor symptoms and urogenital atrophy. HRT should be limited to osteoporosis prevention in women with significant ongoing vasomotor symptoms who are not at an increased risk for cardiovascular disease. An annual, individualized, risk/benefit reassessment should be performed on these patients. Further research is needed to assess the potential risks and benefits of various formulations, combinations, doses, and delivery routes of estrogen for postmenopausal women. PMID- 12734029 TI - Breastfeeding and postmenopausal osteoporosis. AB - Bone loss associated with osteoporosis occurs with high frequency among the elderly and often results in debilitating fractures. A combination of lifestyle behaviors, genetic predisposition, and disease processes contributes to bone metabolism. Therefore, any discussion regarding bone health must address these factors. The impact of menopause on bone turnover has been generally well studied and characterized. Breastfeeding places significant stress on calcium metabolism and, as a consequence, directly influences bone metabolism. The most significant factors affecting bone mineral density (BMD) and bone metabolism are the duration and frequency of lactation, the return of menses, and pre-pregnancy weight. Although transient, lactation is associated with bone loss. As clinical guidelines and public health policies are being formulated, there is a compelling need for further investigation into the relationship of lactation, BMD, and subsequent risk of osteoporosis. Better understanding of this relationship will provide new opportunities for early intervention and ultimately help in the prevention of bone loss in postmenopausal women. PMID- 12734028 TI - Bones and nutrition: common sense supplementation for osteoporosis. AB - Osteoporosis is a serious public health concern. Skeletal fragility, leading to spine and hip fractures, is a major source of morbidity and mortality. Adequate calcium intake from childhood to the end of life is critical for the formation and retention of a healthy skeleton. It is important to prevent bone loss from occurring, to identify potential risk factors, and to correct them. Many genetic and lifestyle factors influence the risk for osteoporosis. Among these, diet is believed to be one of the most important, especially the roles of calcium and vitamin D. Deficiency in other dietary factors--eg, protein, vitamin K, vitamin A, phytoestrogens, and other nutrients--might also contribute to the risk for osteoporosis. In this article, the roles of diet and nutritional supplementation in preventing and treating osteoporosis are reviewed. PMID- 12734030 TI - Identifying the osteopenic patient and preventing worsening of the disease. AB - Osteopenia is a progressive condition that places patients at risk for increased morbidity and mortality if untreated. Patients with bone loss of at least 1.0 standard deviation (SD) from normal are considered osteopenic, whereas those with bone loss of at least 2.5 SDs are considered osteoporotic. Osteoporotic patients are treated to stop bone loss and restore bone mineral density. Patients with bone loss in the osteopenic range, even osteopenic patients with additional risk factors, might not receive treatment. National Osteoporosis Foundation guidelines recommend treatment when bone loss is 2.0 SDs, or less when additional risk factors are present. This is supported by the literature; treatments for bone loss are approved for the prevention of osteoporosis. PMID- 12734031 TI - Comparing screening methods for osteoporosis. AB - Recently, much has been published about osteoporosis and the suspected vast numbers of patients who are undiagnosed or at risk. Various groups, including the US Preventative Services Task Force and The National Osteoporosis Foundation, have attempted to highlight the recommendations regarding who and when to screen. We know that for a screening test to be effective, not only must it predict morbidity far enough in advance that something can be done but it also must be widely available and cost effective. There are several methods of screening with varying sensitivity and specificity for identifying people at risk for osteoporotic fracture. After an examination of all available approved testing methods, it seems that dual-energy x-ray absorptiometry (DEXA) and calcaneal ultrasound best predict patients at risk for fracture. However, because of the length of time needed to demonstrate bone mineral changes and the small magnitude of these changes, DEXA seems to be the most cost-effective method to follow patients who are receiving treatment. PMID- 12734032 TI - Challenges in treating post-traumatic stress disorder and attachment trauma. AB - Treating women suffering from trauma poses significant challenges. The diagnostic prototype of post-traumatic stress disorder (PTSD) is based on single-event trauma, such as sexual assault in adulthood. Several effective cognitive- behavioral treatments for such traumas have been developed, although many treated patients continue to experience residual symptoms. Even more problematic is the complex developmental psychopathology stemming from a lifetime history of multiple traumas, often beginning with maltreatment in early attachment relationships. A history of attachment trauma undermines the development of capacities to regulate emotional distress and thereby complicates the treatment of acute trauma in adulthood. Such complex trauma requires a multifaceted treatment approach that must balance processing of traumatic memories with strategies to contain the intense emotions this processing evokes. Moreover, conducting such treatment places therapists at risk for secondary trauma such that trauma therapists also must process this stressful experience and implement strategies to regulate their own distress. PMID- 12734033 TI - Outpatient management of eating disorders in adults. AB - Eating disorders are prevalent in the young adult female population. Given the serious medical, nutritional, and psychological risks associated with eating disorders, it is advisable that patients be seen within the framework of a multidisciplinary team. Psychotherapy is the most effective treatment modality for eating disorders and constitutes the core of mental health treatment. Although cognitive behavior therapy trials dominate the research literature on interventions for bulimia nervosa and binge-eating disorder, various modalities of psychotherapy have efficacy. Active weight management is also a key component of treatment for anorexia nervosa. Psychotropic medication therapy is not generally useful for the primary symptoms of anorexia nervosa, whereas it is moderately effective in the treatment of both bulimia nervosa and binge-eating disorder. Given the patient population at Massachusetts General Hospital, to which patients often present with serious and chronic symptoms, comorbid medical and psychiatric illness, and history of poor response to treatment, we have found a flexible and eclectic treatment approach most useful clinically. PMID- 12734034 TI - Hormonal basis of mood and postpartum disorders. AB - Mood disorders during the postpartum period occur in 10% to 15% of women. The hormonal basis of these disorders, however, has not been investigated systematically and extensively. We review recent studies, primarily from the past 5 years, in which investigators examined the major categories of proposed hormonal etiologies, including gonadal steroids, thyroid hormones, cortisol, prolactin, and melatonin, and then present descriptive statistics of our preliminary findings in these hormonal dimensions from a group of 20 depressed and normal control pregnant and postpartum women. PMID- 12734035 TI - Psychopharmacologic treatment of depression during pregnancy. AB - Increasing numbers of patients are being treated for mood disorders. Most of these patients, particularly those with the diagnosis of major depression, are women of childbearing years. Depression can also occur in the context of bipolar disorder. Concerns regarding fetal exposure to medication, either planned or unplanned, are becoming more pressing in the clinical practices of psychiatrists and primary care physicians. There are relatively few study data available to guide clinicians in the use of psychotropic medications during pregnancy, owing to obvious problems in designing studies of the effects of medications on pregnant women, fetuses, and infants. In this article clinically relevant study and practice information is provided, and suggestions regarding the approach to the treatment of mood disorders during pregnancy based on a risk assessment mode are given. PMID- 12734036 TI - Emerging issues in gender and ethnic differences in substance abuse and treatment. AB - The emerging understanding of gender differences among ethnic minorities in the rates, etiology, course, and treatment of substance abuse and common comorbid mental health disorders has significant scientific and practical implications. Growing recognition of these differences and their implications for treatment and policy decisions has highlighted research gaps and the need for more thoughtful application of the knowledge gained from existing research findings. In this brief review, we outline some of the unique aspects of substance abuse and comorbid mental health problems for women, as well as for women of various ethnic/cultural groups, including specific barriers to obtaining and remaining in treatment. Research challenges to improving limited knowledge about the rates, course, and treatment of substance abuse disorders among ethnic minority women are highlighted. PMID- 12734037 TI - Sexual orientation and women's smoking. AB - An extensive review of the literature on sexual orientation and health, lesbian health, and women and smoking revealed no studies that focus on smoking among lesbians or bisexual women. However, several health surveys conducted in the past 15 to 20 years report rates of current smoking. Findings from these studies as a whole suggest that lesbians are more likely than heterosexual women to smoke. Research on women and smoking is reviewed to identify potential risk factors for lesbians' smoking. Implications for future research and for prevention and intervention are discussed. PMID- 12734038 TI - Stellate cells in the digestive tract. PMID- 12734039 TI - The changing face of esophageal malignancy. AB - Concern has been expressed about the rapid increase in the incidence of esophageal carcinoma in the United States. This rise is due to an increase in the number of cases of adenocarcinoma of the esophagus. Because of the relatively small number of cases of esophageal carcinoma, the absolute risk of developing this cancer in the United States remains small. Potential origins for this increase in esophageal adenocarcinoma are examined in this review, including the risk induced by obesity, low dietary antioxidants, high dietary fat, family history of breast cancer, smoking, gastroesophageal reflux, and Barrett's esophagus. The risk of esophageal adenocarcinoma is inversely associated with infection by Helicobacter pylori organisms. A better understanding of risk factors involved in the increased incidence of esophageal adenocarcinoma is important for development of new preventive strategies for this serious disorder. PMID- 12734040 TI - Surgical management of esophageal malignancy. AB - Esophageal carcinoma is a highly lethal disease with increasing prevalence and an equally dramatic epidemiologic shift. Its causal association with gastroesophageal reflux disease and adenocarcinoma of the esophagus is well established, and the molecular events underlying this progression from mucosal injury to metaplasia to dysplasia to carcinoma are now becoming clear. Current diagnostic modalities and preoperative staging systems have significant limitations. The extent of surgical resection for esophageal carcinoma remains controversial. Disease confined to the mucosa and submucosa is more common, and endoscopic ablative techniques have been proposed. However, preoperative evaluation of tumor depth and regional nodal metastases remains inadequate in these very early lesions and urges caution before adoption of therapies that may compromise cure. Patients with disease confined to the mucosa or submucosa should undergo resectional therapy aimed at removing the entire esophageal wall, including the periesophageal and perihiatal lymph nodes. For disease penetrating the submucosa, the extent of surgical therapy must be tailored to the objectives of treatment (cure vs palliation) and preoperative stage. Although data from seven prospective, randomized trials are encouraging, no clear survival benefit has been documented for neoadjuvant combined-modality therapy. Surgical resection remains the standard of care and best chance for cure in the treatment of esophageal malignancy, with combined-modality therapy reserved for prohibitive surgery candidates. PMID- 12734041 TI - New approach to malignant strictures of the esophagus. AB - Primary esophageal cancer is the most common cause of malignant esophageal stricture. Prognosis and treatment outcomes vary with the stage of the disease. Endoscopic ultrasound has a high accuracy rate for local and regional staging. Surgery is curative for early cancer. Endoscopic mucosal resection, photodynamic therapy, or brachytherapy can be used with curative intent for early cancer, especially in patients with comorbid conditions precluding surgery. Unfortunately, the majority of patients with esophageal cancer present with advanced disease. The primary aim in these patients is to alleviate symptoms with a minimum of side effects and reinterventions. Palliative surgery or chemoradiotherapy can be associated with high morbidity and mortality rates. Several endoscopic techniques for palliation are available, and all have the potential of significantly improving swallowing. The choice of a particular endoscopic approach is usually determined by local expertise and characteristics of the stricture. PMID- 12734042 TI - Endoscopic therapy for gastroesophageal reflux disease. AB - Many new and developing endoscopic techniques for treatment of gastroesophageal reflux disease (GERD) are now or will soon be available for clinical use. These new therapies are directed to the gastroesophageal junction and include three basic techniques, which are outlined in this review: thermal remodeling and neurolysis of the lower esophageal sphincter (LES) zone; bulking or reinforcing of the LES; and plicating gastric folds at the gastric cardia. Only two of these treatments, radiofrequency ablation and fundic fold plication, have been approved by the US Food and Drug Administration (FDA). Published clinical research on endoscopic treatment is reviewed, although it is mostly comprised of open-label trials and anecdotal clinical reports. Complications of endoscopic GERD treatments are described as well as real and potential pitfalls of these techniques. Problems caused by premature introduction of endoscopic GERD treatments into the marketplace are also discussed, along with the rationale for FDA approval of similar devices. PMID- 12734043 TI - Nonsurgical approaches to esophageal malignancy. AB - Esophageal cancer remains one of the leading causes of cancer death worldwide. Patients generally present with progressive dysphagia, malnutrition, and weight loss. The diagnosis commonly involves radiologic studies and conventional esophagogastroduodenoscopy. Advances in endoscopic evaluation have allowed early detection of premalignant and malignant lesions. These techniques include chromoscopy, which can be performed in conjunction with high resolution/magnification endoscopy, and fluorescent endoscopy. Such techniques as endoscopic ultrasound with dedicated echoendoscopes or high-frequency probes, positron emission tomography, optical coherence tomography, endoscopic magnetic resonance imaging, and tactile sensing may complement conventional imaging by CT to enhance staging accuracy. Because the majority of patients present with incurable disease at the time of diagnosis, nonsurgical approaches to their management have evolved. These include endoscopic mucosal resection, stenting, tumor ablation, and palliative chemoradiotherapy. The ablative techniques include argon plasma coagulator therapy, laser, and photodynamic therapy. For patients with early malignancies of the esophagus who are not surgical candidates, such techniques may be used with curative intent. PMID- 12734045 TI - Perspectives on osteoporosis in pediatric inflammatory bowel disease. AB - Osteoporosis is now recognized as a problem in children with chronic illness. Decreased bone mineral density and increased risk of fracture have been reported in children with inflammatory bowel disease (IBD). Recent studies have led to a better understanding of the pathogenesis of bone loss. There are many risk factors for osteopenia and osteoporosis in children with IBD. Dual-energy x-ray absorptiometry remains the diagnostic procedure of choice for assessment of bone mineral density, but other modalities are being explored. Guidelines for diagnosis and treatment of osteoporosis in children have not been established. This article reviews the current understanding of osteopenia and osteoporosis in children with IBD. PMID- 12734047 TI - Pathophysiology and management of pediatric ascites. AB - Ascites accumulation is the product of a complex process involving hepatic, renal, systemic, hemodynamic, and neurohormonal factors. The main pathophysiologic theories of ascites formation include the "underfill," "overflow," and peripheral arterial vasodilation hypotheses. These theories are not necessarily mutually exclusive and are linked at some level by a common pathophysiologic thread: The body senses a decreased effective arterial blood volume, leading to stimulation of the sympathetic nervous system, arginine vasopressin feedback loops, and the renin-angiotensin-aldosterone system. Cornerstones of ascites management include dietary sodium restriction and diuretics. Spironolactone is generally tried initially, with furosemide added if clinical response is suboptimal. More refractory patients require large-volume paracentesis (LVP) accompanied by volume expansion with albumin. Placement of a transjugular intrahepatic portosystemic shunt is reserved for individuals with compensated liver function who require very frequent sessions of LVP. Peritoneovenous shunts are not used in contemporary ascites management. Liver transplantation remains the definitive therapy for refractory ascites. Although treatment of ascites fails to improve survival, it benefits quality of life and limits the development of such complications as spontaneous bacterial peritonitis. PMID- 12734046 TI - Pediatric issues in new therapies for hepatitis B and C. AB - Two antiviral treatments have been approved for hepatitis B virus (HBV) infection by the US Food and Drug Administration (FDA) for use in children: interferon (IFN)-alpha, 6 MU/m(2) three times a week subcutaneously for 6 months, and lamivudine, 3 mg/kg/d orally for 12 months. Twenty-six percent to 58% of children treated with IFN become HBV DNA negative, and up to 38% become negative to hepatitis B e antigen (HBeAg). Lamivudine, a nucleoside analogue that blocks viral replication by inhibition of the HBV polymerase, has been associated with comparable rates of seroconversion of HBeAg to anti-HBe. Loss of surface antigen occurs in less than 5% of patients treated with lamivudine, compared with 3% to 33% in those treated with IFN-alpha. Fifty percent to 65% of children treated with lamivudine clear HBV DNA after 12 months of therapy, but relapse rates have not been clarified. Patients treated with lamivudine develop drug-resistant (YMDD) mutants in the HBV polymerase at the rate of 16% to 32% per year. No treatments for children with hepatitis C virus (HCV) have been approved by the FDA. However, published reports describe treatment with IFN monotherapy and combination therapy with IFN and ribavirin. Trials of PEG-IFN alone or in combination with ribavirin are in progress. Given the lack of data regarding treatment of HCV in children, it is generally agreed among pediatric hepatologists that the optimal treatment is within the context of randomized, controlled trials. PMID- 12734048 TI - Pediatric otolaryngologic manifestations of gastroesophageal reflux disease. AB - In adults, an estimated 4% to 10% of chronic, nonspecific laryngeal disease seen in otolaryngologic clinics is associated with gastroesophageal reflux disease (GERD). Although no such estimates exist in children, many investigators have reported extraesophageal manifestations of GERD, of which the most common is the association of GERD with asthma and chronic cough. A variety of signs and symptoms of otolaryngologic disease also have been attributed to GERD, including hoarseness, laryngitis, chronic rhinitis, sinusitis, globus pharyngeus, recurrent croup, laryngomalacia, stridor, subglottic stenosis, otalgia, vocal cord granulomas, and oropharyngeal dysphagia. However, proof of the association between these manifestations of otolaryngologic disease and GERD is sparse. Furthermore, the manifestations of otolaryngologic disease often occur in the absence of such classic systems of GERD as heartburn or chest pain. This review explores the role of GERD in otolaryngologic disease in children. PMID- 12734050 TI - Analysis of DNA adducts from chemical carcinogens and lipid peroxidation using liquid chromatography and electrospray mass spectrometry. AB - The identification and dosimetry of DNA adducts are cornerstones of research on cancer etiology in experimental animals and humans. DNA adducts can result from exposure to exogenous chemical carcinogens or through reactions with endogenous by-products of oxidative metabolism. An important research need is high throughput methodology for quantification of any and all adducts that are present at trace amounts in DNA derived from target tissues of animals and humans. This review describes some recent progress made through applications of liquid chromatography coupled with mass spectrometry to structural characterization of unknown DNA adducts and highly sensitive quantitative analysis of target adducts. PMID- 12734049 TI - Nonalcoholic steatohepatitis in children. AB - Nonalcoholic steatohepatitis (NASH) is one entity in a spectrum of chronic liver disease related to obesity, hyperinsulinemia, insulin resistance, and liver cell injury from free fatty acid toxicity or other oxidant stress. The more inclusive term "nonalcoholic fatty liver disease" (NAFLD) is increasingly being used to encompass the entire spectrum, which includes simple hepatic steatosis without inflammation (which may not lead to progressive liver injury), NASH itself, and the resulting cirrhosis (which may be devoid of steatosis). Children get NAFLD, and the incidence of this pediatric liver disease is rising as childhood obesity becomes increasingly prevalent. Although much remains to be learned about pediatric NAFLD, it is already evident that children with NASH risk progressive liver damage, including cirrhosis. Liver biopsy is required for definitive diagnosis, and other causes of fatty liver in childhood must be excluded. Gradual weight loss through increased regular exercise and a low-fat, low-refined carbohydrate diet appears to be effective. Drug treatments are being developed. Pediatric NASH is a serious complication of childhood obesity. PMID- 12734052 TI - Cancer incidence and socioeconomic geography of Finland: a correlation study. AB - The influence that social determinants exert on cancer incidence and patterns is an expected aftermath of the predominantly environmental origin of cancer. This happens because social changes determine a wide range of individual behaviors, and hence of "proximate" causes of cancer. Whereas most of the previous social epidemiological studies focused on individual social classes and groups, in this paper we studied the influence of the social factors on cancer etiology at the larger scale of a whole country: Finland. The ecological analysis of cancer incidence and patterns in the Finnish regions pointed to both local and general correlations between tumors and socioeconomic descriptors. The most important correlations were: a) a general effect of economic and societal factors on cancer (namely, on the profiles of tumors in males and females, on the global cancer incidence in females, and on the difference in cancer incidence between the two sexes); b) a very specific pattern of correlations for the Ahvenanmaa islands; c) a clear geographic cline in tumor profile variability. The ecological epidemiology approach can provide important clues to public health policies. PMID- 12734051 TI - Economic status, smoking, occupational exposure to rubber, and lung cancer: a case-cohort study. AB - Recent studies tend consistently to confirm the presence of a moderate excess risk of lung cancer in the rubber industry. However, the agent responsible for the excess of lung cancer is still obscure. Also, analyses without regard to the modifying effects of sex, economic status, and smoking habit are less than satisfactory. To explore these questions, we have conducted a case-cohort study using the data of 51 lung cancer deaths in 1973-1997 and a random sample (sub cohort) of 188 from among 1598 subjects in a rubber factory in Shanghai, China. We computed the risks of lung cancer by economic status, smoking habit, coal fumes in home, and year of first employment. We assessed lung cancer risks for occupational exposures, unadjusted and adjusted for economic status and smoking. After confounding effects of smoking and economic status were controlled, we found that rate ratios were 1.43 (95% confidence interval (CI) 0.43-4.69), 1.79 (95% CI 0.64-5.03), and 3.76 (95% CI 1.44-9.86) for 1-14, 15-29, and 30-45 exposure-years in curing department, respectively. The data showed significant trends in increased risk of lung cancer with duration of exposure in tire-curing department (score test for trend:, P = 0.004). However, in front rubber processing (weighing and mixing, calendering, extruding, and milling), no significant excess risk of lung cancer was found. If it can be confirmed that nitrosamines are mainly generated in back rubber processing (curing and vulcanizing), it would be reasonable to conclude that excess risk of lung cancer in rubber industry is attributable, at least partially, to exposure to nitrosamines. PMID- 12734053 TI - The measurements of ambient particulates (TSP, PM2.5, PM2.5-10), chemical component concentration variation, and mutagenicity study during 1998-2001 in central Taiwan. AB - During June 1998 and February 2001, the experiments of this study were conducted at four sampling sites (THUPB, THUC, HKIT and CCRT) with different characters (suburban, rural and traffic). The chemical components (Cl-, Na+, K+, Mg2+, Ca2+, Fe, Zn, Pb, Ni) in suspended particle were also analyzed simultaneously. The particulate mass concentrations are higher in the traffic site (CCRT) than the other sampling sites in this study. This is because that high traffic density flow characterized CCRT sampling site. Besides, the fine particle (PM2.5) concentration was the dominant species out of the total suspended particles in central Taiwan, Taichung. The same phenomenon is found in most of cities around the world. Moreover, chloride, nitrate sulfate and ammonium are higher in Taiwan than other sampling sites in the world. The results also indicated that the control of acidic and secondary aerosol pollutants have become an important issue in Taiwan. In addition, mutagenic assays on the organic extracts of airborne particulates at different sampling sites were also conducted in central Taiwan. The data obtained here also reflected that the mutagenicity of the suspended particulates are significantly higher in winter period than it occurred in summer period. PMID- 12734054 TI - Dual role of organosulfur compounds in foods: a review. AB - Organosulfur compounds present in natural food are generally considered as beneficial for health because of their antioxidant and anticarcinogenic properties. This has led to their excessive and long-term consumption. However, there is also evidence that these compounds demonstrate toxicity and adverse health effects suggesting their potential dual biological roles. Thus, they can act as double-edged biological swords. PMID- 12734056 TI - An enriched mixture of trans-10,cis-12-CLA inhibits linoleic acid metabolism and PGE2 synthesis in MDA-MB-231 cells. AB - Conjugated linoleic acid (CLA) isomers are potent inhibitors of mammary tumor cell growth. Evidence suggests that CLA modulates essential fatty acid (EFA) metabolism; however, it is not clear which parts of this pathway are important regulatory points modulated by CLA. Enriched mixtures of D9-cis,11-trans (D9c,11t)- and D10-trans,12-cis (D10t,12c)-18:2 were used to assess outcome measures of EFA metabolism pertaining to membrane phospholipid incorporation, tumor cell growth, and prostaglandin E2 (PGE2) synthesis in the MDA-MB-231 mammary tumor cell line. Tumor cells were treated with linoleic acid (LA), an equal mixture (Mix), or enriched preparations of D9c,11t- or D10t,12c-18:2. Treatment with Mix or the enriched mixture of D10t,12c-18:2 significantly inhibited the synthesis of arachidonic acid (AA) from LA, resulting in increased levels of LA and decreased levels of AA in membrane phosphatidylcholine and phosphatidylethanolamine (P < 0.05). LA and AA levels were not altered in cells treated with enriched D9c,11t-18:2 and were similar to those in LA control treated cells. All CLA treatments reduced [3H]thymidine uptake, an indicator of tumor cell growth, by more than one-half relative to LA controls. MDA-MB-231 cells challenged with AA in the presence of all CLA mixtures resulted in significantly reduced PGE2 synthesis relative to controls treated with LA (P < 0.05). It is evident that individual isomers exert inhibitory effects at specific steps of EFA metabolism, which correspondingly leads to a reduction in PGE2 synthesis and, ultimately, tumor growth. PMID- 12734057 TI - Diet and oral, pharyngeal, and esophageal cancer. AB - Cancers of the upper digestive tract, including those arising in the oral cavity, pharynx, and esophagus, present a significant public health problem worldwide. These cancers are associated with high morbidity and mortality, and identification of protective factors is very important. A number of epidemiological studies have examined the association between vegetables, fruits, carotene, vitamin A, vitamin C, and vitamin E and oral, pharyngeal, and esophageal cancers. The results of 35 epidemiological studies, including one prospective cohort study, one nested case-control study, two randomized controlled trials, nine population-based case-control studies, and 22 hospital based case-control studies, in addition to in vitro and animal studies, were examined to determine whether the criteria for causal assumption were satisfied for a protective role of these dietary components against development of oral, pharyngeal, and esophageal cancers. There is enough evidence to point to a preventive role of vegetable intake, including green vegetables, cruciferous vegetables, and yellow vegetables, total fruit intake, and citrus fruit intake. Yellow fruits are likely to be protective. Carotene, vitamin C, and vitamin E are protective, most likely in combination with each other and other micronutrients. The role of vitamin A is not clear because of conflicting findings in the studies reviewed. PMID- 12734058 TI - A meta-analysis of published literature on waist-to-hip ratio and risk of breast cancer. AB - Epidemiological studies have identified body weight as a risk factor for breast cancer. Beyond the amount of adipose tissue a woman has, its distribution, particularly abdominally, may be a risk factor in breast cancer etiology. Body fat distribution is commonly measured by a waist-to-hip circumference ratio lpar;WHR). We performed a meta-analysis to summarize the published literature on WHR and breast cancer risk. After assembling all published studies, we extracted mean WHRs for study participants and adjusted risk estimates comparing highest with lowest partition of WHR and calculated weighted mean differences in WHR between cases and noncases and summary risk estimates based on study design and menopausal status. The weighted mean difference was 0.016 [95% confidence interval (CI) = 0.005-0.028] for all studies combined. The summary risk estimates were 1.80 (95% CI = 1.29-2.50) for case-control studies and 1.27 (95% CI = 1.07 1.51) for cohort studies. By menopausal status, the summary risks were 1.79 (95% CI = 1.22-2.62) for premenopausal women and 1.50 (95% CI = 1.10-2.04) for postmenopausal women. For all studies combined, the summary risk was 1.62 (95% CI = 1.28-2.04). This meta-analysis indicates that a greater WHR is associated with increased risk of breast cancer and suggests that the avoidance of abdominal obesity may reduce risk of the disease. PMID- 12734059 TI - High-folate diets and breast cancer survival in a prospective cohort study. AB - Recent evidence suggests that adequate dietary folate may attenuate the risk of breast cancer associated with intake of alcohol. However, patients with breast cancer have been commonly treated with antifolate chemotherapies. The present analysis was performed to test the hypothesis that high folate intake may diminish the effectiveness of chemotherapy and, therefore, adversely influence survival. Women at risk of postmenopausal breast cancer (n = 37,105) participated in the Iowa Women's Health Study. Total folate intake (diet + supplements) was estimated from a food frequency questionnaire administered at baseline in 1986 and categorized into tertiles. From all incident breast cancer cases ascertained in the cohort, we selected those with a diagnosis between 1986 and 1994, chemotherapy as first course of treatment, and adequate diet assessment. Mortality was determined through the State Health Registry of Iowa and the National Death Index. Cox regression was used to estimate survival while adjusting for important covariates. Through 14 yr of follow-up, 80 deaths occurred among the 177 breast cancer cases treated with chemotherapy. Among these patients, high folate intake was not associated with worse survival. After adjustment for age, extent of disease, total calories, alcohol, and estrogen receptor status, women with total folate intake in the highest tertile had a mortality risk ratio of 0.88 (95% confidence interval = 0.44-1.76) compared with cases in the lowest tertile of folate. These findings, although preliminary, afford some reassurance that folate supplementation is unlikely to have a significant adverse effect on survival after chemotherapy for breast cancer. PMID- 12734060 TI - Diet as risk for lung cancer: a Swedish case-control study. AB - A case-control study was undertaken to study lung cancer in relation to dietary habits, occupational exposure, and living in urban or country areas. Suspect lung cancer cases in West Sweden and population controls were interviewed using a food frequency questionnaire. The study comprised 177 female and 359 male cases and 916 controls. The cases mainly comprised former and current smokers (82% female, 95% male). For the analysis, cases were divided into the histological diagnoses adenocarcinoma and squamous cell, small cell, and adenosquamous cell carcinomas, as well as into smoking categories. A high frequency of consumption of vegetables was significantly related to a lower risk for adenocarcinoma and squamous cell and adenosquamous cell carcinoma among men and adenocarcinoma among women. A low odds ratio in the highest quartile of vegetable consumption in men was seen in all smoking categories. There were no significant protective effects from fruit in the different lung cancer subgroups, although a significant trend was found for heavy-smoking females. A high consumption of milk was related to an increased risk for lung cancer, especially adenosquamous cell carcinoma. The results suggest that the protective effect or risk due to dietary factors may affect different forms of lung cancer. The results from this as well as previous studies suggest a complex interaction between diet and lung cancer risk, involving the types of lung cancer as well as consumption patterns in the population. PMID- 12734061 TI - Nutritional status in childhood malignancies. AB - In children affected by tumor, nutritional status is important to sustain aggressive chemotherapy and to support normal growth during and after therapy. The aim of this study was to investigate the prevalence of nutritional status disorders in a sample of pediatric oncology day-hospital patients. We measured weight and height in patients affected by solid tumors on or off therapy at short term follow-up (1-24 mo). The study was performed at a pediatric oncology day hospital over a period of 20 consecutive days. A suitable computer package was used to estimate relative body weight (%RBW) and body mass index (BMI) for each patient. Thereafter, the same sample was divided into four weight classes (underweight, normal weight, overweight, and obese) according to %RBW and BMI. Moreover, patients were divided into two groups: on and off therapy. In the off therapy group, no patient was underweight; in the on-therapy group, 26.3% and 15.8% of patients were underweight (not significant) according to %RBW and BMI, respectively. The prevalence of overweight (overweight ++obese) according to %RBW was 36.9% in the on-therapy group and 52.9% in the off-therapy group (P < 0.05); whereas the prevalence of overweight according to BMI was 21% in the on-therapy group and 35.3% in the off-therapy group (P = 0.05). These preliminary data suggest that, in pediatric oncology, nutritional assessment is required to provide nutritional strategies in on-therapy patients whose underweight status prevalence is impressive or in off-therapy children in whom the causes of overweight should be explored. PMID- 12734063 TI - Effect of moderate caloric restriction and/or weight cycling on mammary tumor incidence and latency in MMTV-Neu female mice. AB - Recently, we reported that intermittent caloric restriction-refeeding reduces mammary tumor (MT) incidence and extends latency in murine mammary tumor virus (MMTV)-transforming growth factor (TGF)-a mice to a greater extent than does chronic caloric restriction. Here, this same weight-cycling protocol was applied to MMTV-Neu female mice, which develop MTs at a much younger age than do TGF-a mice. This study consisted of three experimental groups: mice fed an AIN-93M diet ad libitum, mice intermittently fed an AIN-93 modified diet (2-fold increase in protein, fat, vitamins, and minerals) at 50% of the amount fed to the ad libitum fed mice for 3-wk intervals and then fed an AIN-93M diet ad libitum for 3-wk intervals, and mice chronically restricted, pair fed to the intermittently restricted mice by feeding 2:1 mixtures of AIN-93M-AIN-93 modified diets for each 6-wk feeding interval. Mice were euthanized when MTs reached a length of 20 mm or at 80 wk of age. Cumulative caloric intake was 10% lower (not significant) for intermittently restricted mice and 16% lower (P < 0.05) for chronically restricted mice than for ad libitum-fed mice. Final body weights were significantly different as follows: ad libitum-fed > intermittently restricted > chronically restricted. Fat pad weights were greater in ad libitum-fed than in intermittently restricted and chronically restricted mice. MT incidence of ad libitum-fed mice was 37% compared with 22% for intermittently restricted mice and 33% for chronically restricted mice (not significant). There were no differences in MT weight or number among the groups. These results indicate that intermittent caloric restriction-refeeding provides a moderate protective effect, whereas chronic caloric restriction provides no significant protection against MT development in transgenic Neu mice. PMID- 12734062 TI - Circulating concentrations of "free" leptin in relation to fat mass and appetite in gastrointestinal cancer patients. AB - Recent studies have suggested that circulating concentrations of leptin might play a role in cancer cachexia. In the first part of the study, we compared circulating concentrations of free and total leptin, percent fat mass, and the inflammatory markers C-reactive protein (CRP) and interleukin-6 (IL-6), together with appetite score, in age- and gender-matched healthy controls (n = 11) and advanced gastrointestinal cancer patients (n = 26). In the second part of the study, the same measurements were repeated before and after megestrol acetate treatment of weight-losing gastrointestinal cancer patients (n = 10). Body mass index and percent fat mass were significantly lower (P < 0.05) and IL-6 and CRP were significantly higher (P < 0.05) in cancer patients than in controls. There was no difference in the percentage of leptin bound in the circulation between controls and cancer patients. Circulating "free" leptin concentrations correlated with percent fat mass in controls (r = 0.745, P = 0.008) and cancer patients (r = 0.600, P = 0.001). In cancer patients, circulating leptin concentrations, either free or total, were not correlated with IL-6 or CRP concentrations. When adjusted for fat mass, the circulating concentrations of free and total leptin were significantly lower in the cancer patients (P < 0.01). Megestrol acetate treatment significantly increased circulating free and total leptin concentrations in the cancer patients (P < 0.05). There was a significant positive correlation between the change in circulating concentrations of free and total leptin and the change in percent fat mass (r = 0.685, P < 0.05 and r = 0.661, P < 0.05, respectively). The results of the present study indicate that the proportions of free and bound leptin in the circulation do not differ between normal subjects and patients with gastrointestinal cancer and in both groups are related to fat mass. Furthermore, the increase in circulating leptin concentrations after megestrol acetate treatment is not associated with any alteration in leptin binding. PMID- 12734064 TI - Adequate intakes of vitamin E and protein prevent increases of oxidative damage to DNA, lipids, and protein induced by total body irradiation in mice. AB - We examined the influence of the level of dietary protein or vitamin E (VE) on oxidative damage to DNA, lipids, and protein in the liver after total body irradiation (TBI) with X-rays at 1 or 4 Gy. Levels of 8-hydroxydeoxyguanosine, thiobarbituric acid-reactive substances, and protein carbonyls in the liver did not differ among the groups that did not receive TBI. However, oxidative damage to lipids and protein was increased by TBI only in the 1% protein group. DNA damage, lipid peroxidation, or protein oxidation in the liver was increased by TBI in a dose-dependent manner, and the damage was consistently higher in the 1% than in the 20% protein group. In the 1% protein group, a greater decrease in relative spleen weight by TBI was also observed. Concentrations of antioxidants (vitamins C and E and glutathione) in the liver were lower and the concentration of nonheme iron in the liver was higher in the 1% than in the 20% protein group. Mice fed a 1% protein diet became susceptible to TBI-induced oxidative damage, and decreases in antioxidant levels and an increase in iron level were involved in the mechanism of this susceptibility. These results suggest that dietary VE and protein can prevent oxidative damage to DNA, lipid, and protein in mice subjected to TBI. Consumption of a VE-free diet significantly increased 8 hydroxydeoxyguanosine levels in DNA from mice fed the 1% protein diet with TBI, but such changes were not detected in DNA from mice fed the 20% protein diet. PMID- 12734065 TI - Dietary docosahexaenoic acid levels influence the outcome of arabinosylcytosine chemotherapy in L1210 leukemic mice. AB - The purpose of this study was to investigate whether dietary supplementation with the n-3 fatty acid docosahexaenoic acid (DHA) in combination with arabinosylcytosine (AraC) chemotherapy could prolong the life expectancy of mice bearing L1210 leukemia. The four control diets included rodent chow, a diet containing 5% of a blended oil mimicking the fatty acid composition of rodent chow, and diets containing 5% or 10% fat with safflower oil as the main oil source. The two DHA-supplemented diets provided 1.5% or 3.5% DHA and 5% or 10% total fat, respectively. After tumor cell inoculation, mice were treated with AraC for 10 days. Mice fed the 5% safflower oil diet (30.1 -/+ 4.1 days), but not those fed the 10% safflower oil diet, survived longer than the chow-fed animals (22.1 -/+ 3.1 days, P = 0.05). The 1.5%-/+ DHA diet (average intake 1.8 g DHA/kg/day) was associated with a longer life span (33.3 -/+ 3.4 days, P < 0.01 vs. chow-fed) and no incidence of death due to drug toxicity. Further increasing DHA intake (4.5 g DHA/kg/day) resulted in shortened survival time (26.5 -/+ 2.0 days), increased circulating tumor cell burden, and lowered red blood cell concentrations. These data suggest that a modest level of dietary DHA or linoleic acid supplementation may improve the antineoplastic efficacy of AraC. However, overconsumption of DHA reverses the beneficial effect of DHA intake on drug sensitivity. PMID- 12734066 TI - Antitumor activity of milk kefir and soy milk kefir in tumor-bearing mice. AB - The effects of oral administration of milk and soy milk kefirs on tumor growth in tumor-bearing mice and the mucosal immunoglobulin A response in mice were studied. Oral administration of milk and soy milk kefirs to mice inoculated with sarcoma 180 tumor cells resulted in 64.8% and 70.9% inhibition of tumor growth, respectively, compared with controls. In addition, oral administration of the two kefir types induced apoptotic tumor cell lysis. Total immunoglobulin A levels for tissue extracts from the wall of the small intestine were also significantly higher for mice fed a milk kefir or a soy milk kefir regimen for 30 days. These results suggest that milk and soy milk kefirs may be considered among the more promising food components in terms of cancer prevention and enhancement of mucosal resistance to gastrointestinal infection. PMID- 12734067 TI - Food-borne radiolytic compounds (2-alkylcyclobutanones)may promote experimental colon carcinogenesis. AB - Food irradiation is acknowledged as a safe process to improve food quality by reducing microbial contamination. Information on the toxicological potential of 2 alkylcyclobutanones (2-ACBs), radiolytic derivatives of triglycerides found exclusively in irradiated food, is scarce. Wistar rats received daily a solution of highly pure 2-tetradecylcyclobutanone (2-tDCB) or 2-(tetradec-5-enyl) cyclobutanone (2-tDeCB) at a concentration of 0.005% in 1% ethanol as drinking fluid, while control animals received 1% ethanol. All animals received a single intraperitoneal injection of the chemical carcinogen azoxymethane (AOM) at Weeks 3 and 4. At 3 mo after AOM injection, no significant changes were observed in the total number of preneoplastic lesions in the colon of AOM controls and 2-ACB treated animals. After 6 mo, the total number of tumors in the colon was threefold higher in the 2-ACB-treated animals than in the AOM controls. The colon of four of six AOM control rats exhibited only one small tumor ( &6 mm3). Multiple tumors were observed in four and three of six animals treated with 2 tDCB or 2-tDeCB, respectively. Medium (6 < S < 25 mm3) and larger (>25 mm3) tumors were detected only in 2-ACB-treated animals. This is the first demonstration that a compound found exclusively in irradiated dietary fats may promote colon carcinogenesis in animals treated with a chemical carcinogen. PMID- 12734070 TI - Current issues in the identification, assessment, and management of concussions in sports-related injuries. AB - The recent literature has focused on the need for appropriate identification, assessment, and management of sports-related concussion. This article addresses current issues in the prevalence and assessment of sports-related concussion. Despite a paucity of research on female athletes and youth athletes, there is evidence that female athletes are at higher risk for injury than males and that concussions may affect children and young adolescents differently than older adolescents and adults. Sideline, baseline, and postconcussion assessments have become prevalent in documenting preinjury and postinjury performance, tracking recovery rates, and assisting return-to-play decisions. New computerized assessment procedures are growing in popularity and use. Future directions in the assessment and management of sports-related concussion include increased research on prevalence rates and effects of concussions for females and youth athletes, educating parents of youth athletes as well as family physicians on the importance of baseline and postconcussion cognitive assessments, and further validation of computerized assessment measures. PMID- 12734071 TI - Epidemiological considerations of concussions among intercollegiate athletes. AB - The purpose of this study was to examine epidemiological trends of concussions among 15 different intercollegiate sports during the 1997-1998, 1998-1999, and 1999-2000 seasons. Data were collected using the National Collegiate Athletic Association (NCAA) Injury Surveillance System (ISS). For the 15 sports studied during the 3 academic years, the NCAA ISS documented 3,535 team-seasons, 40,547 reportable injuries, 5,566,924 practice athlete exposures (AEs), and 1,090,298 game AEs. Concussions accounted for 6.2% of all reported injuries during this 3 year study. Of all the reported injuries, women lacrosse players (13.9%) reported the highest percentage of suffering a concussion during a game followed by women's soccer (11.4%), men's ice hockey (10.3%), men's lacrosse (10.1%), football (8.8%), women's basketball, (8.5%), field hockey (7.2%), men's soccer (7.0%), wrestling (6.6%), men's basketball (5.0%), baseball (4.2%), and women's volleyball (4.1%). Female athletes from all 7 sports were found to be at a lower risk for suffering concussions during practice sessions than the 8 male sports. However, female athletes were found to be at a greater risk for suffering concussions during games compared to male athletes. Injury trends over the 3- year period indicate concussions continue to be on the rise for athletes participating in collegiate football, men's soccer, and women's and men's basketball. PMID- 12734068 TI - Resveratrol induces downregulation in survivin expression and apoptosis in HTLV-1 infected cell lines: a prospective agent for adult T cell leukemia chemotherapy. AB - Resveratrol, a phytoalexin found in grapes and wine, has been shown to exhibit a wide range of pharmacological properties and is believed to play a role in the chemoprevention of human cancer. Resveratrol has also been shown to induce antiproliferation and apoptosis of several leukemia cell lines. In the present study, we investigated the effect of resveratrol in adult T cell leukemia. Our present observations showed that resveratrol induced growth inhibition in all five human T cell lymphotrophic virus-1-infected cell lines examined, with 50% effective dose of 10.4-85.6 mM. In the resveratrol-treated cells, induction of apoptosis was confirmed by annexin V-based analyses and morphological changes. The most surprising observation was that resveratrol treatment resulted in a gradual decrease in the expression of survivin, an antiapoptotic protein, during cell apoptosis. These findings indicate that resveratrol inhibits the growth of human T cell lymphotrophic virus-1-infected cell lines, at least in part, by inducing apoptosis mediated by downregulation in survivin expression. In view of the accumulating evidence that survivin may be an important determinant of a clinical response in adult T cell leukemia, our present findings have led to the suggestion that resveratrol, a common constituent of the human diet, merits further investigation as a potential therapeutic agent for this incurable disease. PMID- 12734072 TI - The neuropsychology of repeated 1- and 3-meter springboard diving among college athletes. AB - This study examined the neuropsychological effects of repeated springboard diving. It was hypothesized that the impact velocity, which can range from 20 to 30 mph, and accompanying deceleration in the water may lead to concussions and affect the diver's cognitive function. Six varsity National Collegiate Athletic Association Division 1 springboard divers participated in the study. Each diver performed a total of 50 practice dives from either the 1- or 3-m springboard. After each set of 10 dives, the participants were immediately evaluated at poolside using the Symbol Digit Modalities Test, the Stroop Color Word Test, and the Trail Making Test B. Baseline testing revealed, consistent with their athletic specialty, clear neurocognitive strengths among the divers on tests sensitive to proprioception, motor speed, and visual-spatial organization. Results from the serial assessments indicated no detectable neuropsychological deficits among competitive divers compared to baseline testing. Skilled diving at the collegiate level appears to be a safe sport and water appears to present the perfect medium for gradual deceleration. More studies, however, are warranted for 5-, 7.5-, and 10-m platform diving since the impact velocity of the diver from these heights is higher. PMID- 12734073 TI - Recency and frequency of soccer heading interact to decrease neurocognitive performance. AB - This study investigated the role of heading recency interacting with heading frequency in determining neuropsychological deficits associated with heading the ball during soccer play. Sixty-four high-ability male soccer players ages 16 to 34 completed the California Verbal Learning Test (CVLT), the Trailmaking Test, the Paced Auditory Serial Addition Test (PASAT), the Facial Recognition Test, the Rey-Osterrieth Complex Figure, and the Shipley Scales. Heading recency interacted with heading frequency, such that players with the highest self-reported estimates of heading who also experienced heading within the previous 7 days scored significantly lower on CVLT, Shipley, Trailmaking, and PASAT than other combinations of heading and recency. Although strict ball-to-head contacts could not be isolated as sufficient to cause this interaction, these results increase the weight of evidence that heading behavior is problematic for causing at least transient cognitive impairment. PMID- 12734074 TI - Computer-based assessment of sports-related concussion. AB - Sports-related concussion has received considerable attention from neuropsychologists, athletic trainers, team coaches, physicians, families, and athletes. In this context, researchers have recently developed computer programs for the assessment of sports-related concussion. Computer-based assessment of sports-related concussion saves time, allows for team baseline testing, and can be easily incorporated into the sports medicine environment. This article reviews the advantages and limitations of computer-based assessment of sports-related concussion. Within a well-coordinated concussion management program that includes input from a neuropsychologist, computer-based assessment of sports-related concussion will soon be the most common approach for assessing concussion in athletes. PMID- 12734075 TI - Return to play following sports-related mild traumatic brain injury: the role for neuropsychology. AB - Cerebral concussions frequently occur at all levels of athletic competition. The effects from these concussions can be transient or may lead to chronic, debilitating symptoms. A growing literature has established that neuropsychological tests are useful in detecting the subtle neurocognitive changes that occur following concussions. The identification of these deficits and subsequent recovery of function can be important components in making return to-play (RTP) decisions. This article describes the emergence of neuropsychology in sports medicine, discusses the context in which RTP decisions are made, outlines factors that are important to RTP decisions, and presents a model that views the RTP decision as a dynamic risk-benefit analysis that involves complex interactions among variables. It is argued that neuropsychology has a unique, but not exclusive, role in the decision making process. Implications for future research are discussed. PMID- 12734077 TI - Testing the water before diving off the cutting edge. PMID- 12734078 TI - Earlier chemotherapy for breast cancer: perhaps too late but still useful. PMID- 12734079 TI - Molecular staging of melanoma: sensitivity, specificity, and the search for clinical significance. PMID- 12734080 TI - Radiofrequency ablation of malignant liver tumors. AB - BACKGROUND: Radiofrequency ablation (RFA) is being used to treat primary and metastatic liver tumors. The indications, treatment planning, and limitations of hepatic RFA must be defined and refined by surgeons treating hepatic malignancies. METHODS: A review of the experience using RFA to treat unresectable primary and secondary hepatic malignancies at the University of Texas M. D. Anderson Cancer Center in Houston, Texas, and the G. Pascale National Cancer Institute in Naples, Italy, is provided. Patient selection, treatment approach, local recurrence rates, and overall cancer recurrence rates following RFA are described. The current literature on RFA of hepatic malignancies is reviewed. RESULTS: RFA of hepatic tumors can be performed percutaneously, laparoscopically, or during an open surgical procedure. Incomplete treatment manifest as local recurrence is more common with a percutaneous approach. The morbidity and mortality rates associated with hepatic RFA are low. Local recurrence rates are low if meticulous treatment planning is performed. RFA can be combined safely with partial hepatic resection of large lesions. The long-term survival rates following RFA of primary and metastatic liver tumors have not yet been established. CONCLUSIONS: RFA of hepatic malignancies is a safe and promising technique to produce coagulative necrosis of unresectable hepatic malignancies. Experience with this treatment modality is not yet mature enough to establish long-term outcomes. PMID- 12734081 TI - Feasibility of adjuvant hepatic arterial infusion of chemotherapy after radiofrequency ablation with or without resection in patients with hepatic metastases from colorectal cancer. AB - BACKGROUND: The safety of combined hepatic artery infusion chemotherapy (HAI) and radiofrequency ablation (RFA) for liver metastases has not been assessed. We conducted a study to determine the feasibility of using HAI after RFA for colorectal cancer (CRC) liver metastases. METHODS: Between 1996 and 2001, patients with hepatic metastases from CRC were enrolled onto a prospective study of RFA plus HAI consisting of continuous-infusion floxuridine and bolus fluorouracil. Surgical complications, treatment-related toxicities, and patient outcomes were recorded. RESULTS: Fifty patients were treated with RFA and HAI with or without resection. A median of two lesions per patient, with a median greatest diameter of 2.0 cm, were treated with RFA. Postoperative complications, including 1 death, occurred in 11 of 50 patients. Toxicity from HAI was relatively mild. At 20 months' median follow-up, 32% of patients remained disease free. Ten percent of patients had recurrences at the site of RFA, 30% developed new liver metastases, and 48% developed extrahepatic disease. CONCLUSIONS: RFA of CRC liver metastases followed by HAI is feasible and is associated with acceptable complication and toxicity rates. The high rate of disease recurrence in our patients indicates that novel combinations of regional and systemic therapies are needed to improve patient outcomes. PMID- 12734082 TI - Prognostic significance of vascular endothelial growth factor, basic fibroblast growth factor, and angiogenin in patients with resectable hepatocellular carcinoma after surgery. AB - BACKGROUND: Hepatocellular carcinoma (HCC) is a hypervascular malignancy. Vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and angiogenin (ANG) are important angiogenic factors of neoangiogenesis. This study investigated the predictive value of serum VEGF, bFGF, and ANG in tumor recurrence, disease-free survival (DFS), and overall survival (OS) in HCC patients. METHODS: Preoperative serum VEGF, bFGF, and ANG were measured in 98 patients with resectable HCC and in 15 healthy controls. The median follow-up time was 43 months. RESULTS: Preoperative serum VEGF was increased in patients with resectable HCC compared with healthy controls (P <.05). Increased serum VEGF was correlated with tumor recurrence (P =.001). Univariate analysis showed that serum VEGF, tumor-node-metastasis stage, tumor size and number, macroscopic portal vein invasion, and microscopic vascular invasion were correlated with OS and DFS. Serum bFGF and ANG were not associated with survival. Multivariate analysis showed that serum VEGF was the most significant predictor of DFS (relative risk, 2.35; 95% confidence interval, 1.26-4.39; P =.007) and OS (relative risk, 3.44; 95% confidence interval, 1.81-6.57; P <.001) in HCC patients after surgical resection. CONCLUSIONS: Preoperative serum VEGF is a significant independent predictor of tumor recurrence, DFS, and OS in patients with resectable HCC. PMID- 12734083 TI - Body mass index does not affect systematic D2 lymph node dissection and postoperative morbidity in gastric cancer patients. AB - BACKGROUND: The extent of standard lymph node dissection (D1, D2, or D3) in gastric cancer patients is still controversial. Several prospective European trials attained contradictory results. A generally increased body mass index (BMI) of the European patients was assumed to be one of the major causes for postoperative morbidity. METHODS: We evaluated the effect of BMI on the quality of routine D2 lymph node dissection and on postoperative morbidity in patients with gastric cancer who underwent a potentially curative total gastrectomy. A total of 199 consecutive gastric cancer patients who underwent a total gastrectomy and a routine D2 lymph node dissection between 1992 and 2001 were included in the study. According to BMI, they were assigned to three groups: group A, with BMI <25 kg/m(2) (normal body weight); group B, with BMI of 25 to 30 kg/m(2) (overweight); and group C, with BMI >30 kg/m(2) (obesity). Parameters such as complete histopathological staging, intraoperative blood loss, length of operation, and surgical and nonsurgical morbidity were recorded and correlated within the different groups. RESULTS: No significant differences were found with regard to the number of examined lymph nodes, blood loss, length of operation, surgical complications, or length of stay in the intensive care unit. CONCLUSIONS: In contrast to comparable Japanese studies, our analysis reveals that even for overweight patients, a standard D2 lymph node dissection is justified without significantly increased morbidity. PMID- 12734084 TI - Randomized multicentric study of perioperative chemotherapy with mitoxantrone in early breast cancer. AB - BACKGROUND: To confirm the hypothesis that reducing the interval between surgery and adjuvant chemotherapy could improve prognosis, a randomized multicentric study of adjuvant perioperative chemotherapy (POC) in breast cancer was initiated. METHODS: A total of 552 patients were randomized to evaluate whether the addition of POC to standard adjuvant treatment significantly improved outcome. Patients were stratified according to menopausal status, with 362 patients in the postmenopausal group and 192 patients in the premenopausal group. Premenopausal women with positive axillary nodes, negative hormonal receptors, or grade 3 tumors received adjuvant mitoxantrone-based chemotherapy. Node-negative premenopausal patients with grade 1 or 2 tumors expressing hormonal receptors received no standard adjuvant treatment. All postmenopausal women received hormonal therapy (tamoxifen 20 mg/day for 3 years). The perioperative regimen was a 14 mg/m(2) mitoxantrone infusion at the end of tumor excision. RESULTS: With a median follow-up of 6.1 years, this study showed no significant advantage of POC on overall survival, disease-free survival, or metastasis-free survival for the total cohort or for the premenopausal and postmenopausal groups. CONCLUSIONS: POC was a safe procedure in this study. However, the addition of POC to standard adjuvant treatment offered no benefit in breast cancer adjuvant treatment. PMID- 12734085 TI - Patterns of recurrence after sentinel lymph node biopsy for breast cancer. AB - BACKGROUND: Sentinel lymph node biopsy (SLNB) is gaining acceptance as an alternative to axillary lymph node dissection. The purpose of this study was to determine the frequency and pattern of disease recurrence after SLNB. METHODS: Two-hundred twenty-two consecutive patients undergoing SLNB from April 6, 1998, to October 27, 1999, and who were >or=24 months out from their procedure were identified from a prospectively maintained database. Retrospective chart review and data analysis were performed to identify variables predictive of recurrence. RESULTS: The median patient follow-up was 32 months (range, 24-43 months). A total of 159 patients (72%) were sentinel lymph node (SLN) negative and had no further axillary treatment. Five of these patients (3.1%) developed a recurrence (one local and four distant), with no isolated regional (axillary) recurrences. Sixty-three patients (28%) were SLN positive and underwent a subsequent axillary lymph node dissection. Six of these patients (9.5%) developed a recurrence (three local, one regional, and two distant). Pathologic tumor size (P <.001), lymphovascular invasion (P =.018), and a positive SLN (P =.048) were all statistically significantly associated with disease recurrence. CONCLUSIONS: With a minimum follow-up of 24 months, patients with a negative SLN and no subsequent axillary treatment demonstrate a low frequency of disease recurrence. This supports the use of SLNB as the sole axillary staging procedure in SLN-negative patients. PMID- 12734086 TI - Magnetic resonance imaging in patients diagnosed with ductal carcinoma-in-situ: value in the diagnosis of residual disease, occult invasion, and multicentricity. AB - BACKGROUND: Although magnetic resonance imaging (MRI) has been shown to be a sensitive imaging tool for invasive breast cancers, its utility in ductal carcinoma-in-situ (DCIS) of the breast remains controversial. We studied the performance of MRI in patients with known DCIS for assessment of residual disease, occult invasion, and multicentricity to determine the clinical role of MRI in this setting. METHODS: Fifty-one patients with biopsy-proven DCIS underwent contrast-enhanced MRI before surgical treatment. Pre-, early post-, and late postcontrast three-dimensional gradient echo images were obtained and MRI findings were correlated with histopathology. When possible, the performance of MRI and mammography was compared. RESULTS: The accuracy of MRI was 88% in predicting residual disease, 82% in predicting invasive disease, and 90% in predicting multicentricity. The performance of MRI was equivalent in the core biopsy group when compared with the surgical biopsy group. For occult invasion only, MRI and mammography were equivalent. However, overall, MRI was more sensitive and had a higher negative predictive value than mammography. CONCLUSIONS: MRI of DCIS can serve as a useful adjunct to mammography by providing a more accurate assessment of the extent of residual or multicentric disease. The performance of MRI is not significantly affected by antecedent surgical excision. MRI may be particularly valuable if preoperatively negative. PMID- 12734087 TI - Mastectomy by inverted drip incision and immediate reconstruction: data from 510 cases. AB - BACKGROUND: Immediate reconstruction of the breast can be performed in selected cases after mastectomy for breast cancer or after prophylactic mastectomy in patients with a high risk of developing breast cancer. Despite the frequency with which these procedures are performed, data from large series of subpectoral implantation of silicone prostheses in combination with a skin-saving approach are lacking. METHODS: In this retrospective study, data on complications and late surgical interventions in 356 patients who underwent 510 mastectomies with an inverted drip incision and immediate reconstruction (MIDIIR) were analyzed to determine potential prognostic factors of early complications. RESULTS: In 82% of the MIDIIRs, the postoperative course was uneventful. In 18%, the complications were infection (32 cases), necrosis of the skin flap (29 cases), bleeding (31 cases), and protrusion of the prosthesis (20 cases), resulting in surgery in 9, 12, 15, and 20 cases, respectively. At the end of the follow-up period, 30 (6%) prostheses were definitively removed. Age, size of the prostheses, radiotherapy, previous lumpectomy, and indication for mastectomy were not significant factors for the prognosis of early complications. CONCLUSIONS: With the right technique and indications, MIDIIR is a very safe procedure and should be one of the surgical treatments that can be offered in the overall management of patients with, or at high risk for, breast cancer. PMID- 12734088 TI - Prognostic significance of reverse transcriptase-polymerase chain reaction negative sentinel nodes in malignant melanoma. AB - BACKGROUND: Polymerase chain reaction (PCR) is a molecular biology technique that can detect a single metastatic cell in 10(6) to 10(7) normal cells. Its use has been proposed as an additional new method for the detection of malignant melanoma nodal metastases in the sentinel lymph node (SLN) to improve the detection rate guaranteed so far by standard histology (hematoxylin and eosin; H&E) and immunohistochemistry (IHC). METHODS: Since October 1995, 137 patients with primary cutaneous melanoma (Breslow thickness,.75-4 mm) have undergone surgery for selective lymphadenectomy. To identify the SLNs, every patient had preoperative lymphoscintigraphy and a vital dye perilesional injection, followed by a gamma probe-guided operation. RESULTS: In 134 patients at least one SLN was detected, with a detection rate of 98%. Every SLN was examined by H&E and IHC (S 100 antigen and HMB-45 protein). The messenger RNA codifying for tyrosinase and MART-1 (melanoma antigen recognized by T cells) was used as the target sequence for the reverse transcriptase (RT)-PCR. The results showed 11% positive SLNs with IHC and H&E examination and 63% with RT-PCR. No recurrence was noted at follow-up in the group with RT-PCR-negative nodes (absence of false-negative cases). CONCLUSIONS: In our experience, RT-PCR SLN negativity is achieving a very favorable prognostic significance. However, RT-PCR positivity is still to be evaluated. Furthermore, results obtained with this method have been shown so far to be independent of Breslow's tumor thickness. PMID- 12734089 TI - Sentinel lymph node biopsy for patients with cutaneous desmoplastic melanoma. AB - BACKGROUND: Although desmoplastic melanoma (DM) often presents at a locally advanced stage, nodal metastases are rare. We describe our experience with lymphatic mapping and sentinel lymph node biopsy (SLNB) in patients with DM to characterize the biological behavior of these tumors. METHODS: Twenty-seven patients with cutaneous DM underwent wide excision and attempted SLNB between 1996 and 2001. All pathology was reviewed by a single dermatopathologist (KB). Clinical and histological features were recorded. RESULTS: There were 20 male and 7 female patients. The median age was 64 years (range, 35-83 years). The head and neck was the most commonly involved anatomical region (n = 14). The median Breslow thickness was 2.2 mm. Twenty-four patients underwent successful SLNB. No patient had a positive sentinel node. At a median follow-up of 27 months, five patients recurred (four systemic and one local); all five had undergone successful SLNB. Two of these patients died of disease, two are alive with disease, and one remains alive and disease free. No patient experienced failure in a regional nodal basin. CONCLUSIONS: DM is a biologically distinct form of melanoma, with a very low incidence of regional lymph node metastases, either at presentation or in long-term follow-up. This biology should be considered when designing rational treatment strategies for these patients. PMID- 12734090 TI - Sentinel lymph node mapping for thick (>or=4-mm) melanoma: should we be doing it? AB - BACKGROUND: Thick (>or=4-mm) primary melanomas are believed to be associated with a high incidence of occult distant metastases. The use of sentinel lymph node (SLN) mapping and biopsy in the treatment lesions has been questioned. METHODS: A retrospective review of a computerized database identified 114 patients who underwent successful SLN mapping and biopsy from January 1, 1994, to December 31, 1999. Records were reviewed for clinicopathologic features of the patients and their tumors. Survival curves were constructed from Kaplan-Meier estimates and analyzed with log-rank tests and Cox proportional hazards modeling. RESULTS: There were 75 men and 39 women with a mean age of 57 years (range, 24-85 years). The primary tumor sites were head and neck (n = 29; 25.4%), trunk (n = 44; 38.6%), and extremities (n = 41; 36%). Tumor thickness ranged from 4 to 17 mm (median, 5.2 mm; mean, 6.3 mm). Ulceration was present in 40 (35.1%) tumors. Thirty-seven patients (32.5%) had a positive SLN biopsy, and 18 of these patients (48.6%) had a single tumor-positive lymph node after dissection. The mean follow up was 37.8 months. The overall 3-year survival for SLN-negative patients was 82%, versus 57% for SLN-positive patients (P =.006). Lymph node status and tumor ulceration were independent predictors of overall survival in multivariate Cox regression analysis. CONCLUSIONS: The pathologic status of the SLN in patients with thick melanomas is a strong independent prognostic factor for survival, and SLN mapping should be routinely performed. PMID- 12734091 TI - Lymphoscintigraphy and sentinel node biopsy accurately stage melanoma in patients presenting after wide local excision. AB - BACKGROUND: Patients have traditionally been considered candidates for sentinel node biopsy (SNBx) only at the time of wide local excision (WLE). We hypothesized that patients with prior WLE may also be staged accurately with SNBx. METHODS: Seventy-six patients, including 18 patients from the University of Virginia and 58 from a multicenter study of SNBx led by investigators at the University of Vermont, who had previous WLE for clinically localized melanoma underwent lymphoscintigraphy with SNBx. Median follow-up time was 38 months. RESULTS: Intraoperative identification of at least 1 sentinel node was accomplished in 75 patients (98.6%). The mean number of sentinel nodes removed per patient was 2.0. Eleven patients (15%) had positive sentinel nodes. Among the 64 patients with negative SNBx, 3 (4%) developed nodal recurrences in a sentinel node-negative basin simultaneous with systemic metastasis, and 1 (1%) developed an isolated first recurrence in a lymph node. CONCLUSIONS: This multicenter study more than doubles the published experience with SNBx after WLE and provides much-needed outcome data on recurrence after SNBx in these patients. These outcomes compare favorably with the reported literature for patients with SNBx at the time of WLE, suggesting that accurate staging of the regional lymph node bed is possible in patients after WLE. PMID- 12734092 TI - Flow cytometric DNA analysis for determination of malignant potential in adrenal pheochromocytoma or paraganglioma: an Indian experience. AB - BACKGROUND: We analyzed the histological features and DNA flow cytometric results in 34 patients with pheochromocytoma and paragangliomas and attempted correlation with the biological behavior for determination of the malignant potential of these tumors. METHODS: DNA analysis was done on a FACSort flow cytometer using paraffin-embedded tissues. Histopathological analysis was performed using parameters, i.e., cell size (large, medium, and small), cell size variation, mitotic rate, nuclear pleomorphism, golden yellow to brown pigment in the tumor, necrosis, and venous invasion. RESULTS: Six tumors had high (>5/10HPF) mitotic rate while venous invasion was seen in three tumors. Fifty percent (18/34) of patients had aneuploid tumors, and 68% (23/34) of patients had high (>10%) S phase fraction tumors. Aneuploidy correlated with >5/10HPF mitotic rate (P <.05) and diploidy with golden yellow to brown pigment (P <.01). The patients with aneuploid tumor had a worse prognosis than patients with diploid tumors (P =.004). No such difference was observed with low and high S-phase fractions (P =.748), presence and absence of venous invasion (P =.927), and mitotic rate (P =.159). Nuclear pleomorphism and necrosis were not significant factors in prognosis. CONCLUSIONS: Flow cytometric DNA analysis of paragangliomas and pheochromocytomas correlated with biological behavior in the patients with regard to metastasis and overall survival in the patients. PMID- 12734093 TI - Histopathologic type: an independent prognostic factor in primary soft tissue sarcoma of the extremity? AB - BACKGROUND: We attempted to define the effect of tumor histotype on local recurrence, distant metastasis, and disease-specific survival in patients with surgically treated primary extremity sarcoma. METHODS: A total of 951 patients with primary, localized soft tissue extremity sarcoma were followed up prospectively. Patient- and tumor-related variables, including histopathologic type, were used to identify independent prognostic factors for the study end points of local recurrence, distant recurrence, and disease-specific survival. RESULTS: There were 137 local recurrences, and significant adverse prognostic factors for local recurrence were patient age >50 years, microscopically positive margins, and malignant peripheral nerve tumor. Adverse prognostic factors for distant recurrence (200 patients) were tumor size >5 cm, tumors beneath the investing fascia, high tumor grade, and leiomyosarcoma. Of the 199 patients who died of disease-related causes, patient age >50 years, tumors beneath the investing fascia, high tumor grade, microscopically positive margin, tumor size >5 cm, leiomyosarcoma, and malignant peripheral nerve tumor were adverse prognostic factors. CONCLUSIONS: These data suggest that differences in biological behavior may exist between sarcoma histotypes and deserve further study. PMID- 12734095 TI - Depth of subserosal invasion predicts long-term survival after resection in patients with T2 gallbladder carcinoma. AB - BACKGROUND: This study aimed to identify a subgroup of patients with inapparent T2 gallbladder carcinoma who may be best suited for radical second resection. METHODS: A retrospective analysis was conducted of 126 patients with pathologic stage T2 (pT2) gallbladder carcinoma (51 with clinically evident tumor and 75 with inapparent tumor). Depth of subserosal invasion was measured histologically in each gallbladder specimen. The median follow-up period was 113 months. RESULTS: In all 126 patients, depth of subserosal invasion was the strongest independent prognostic factor by univariate (P <.0001) and multivariate (relative risk, 9.27; P <.0001) analyses. Among the 75 patients with inapparent tumor, the outcome after resection was significantly better in patients who had undergone radical second resection than in patients who had undergone cholecystectomy alone (P =.0006). When depth of subserosal invasion was divided into 2 mm, the effectiveness of radical second resection remained only in patients with subserosal invasion >2 mm (P =.0004). CONCLUSIONS: Depth of subserosal invasion best predicts postresectional long-term survival of pT2 gallbladder carcinoma patients. Among patients with inapparent pT2 tumors, those with subserosal invasion >2 mm are good candidates for radical second resection. PMID- 12734094 TI - Cauterization versus fibrin glue for aerostasis in precision resections for secondary lung tumors. AB - BACKGROUND: Aerostasis control in multiple resections for metastatic pulmonary diseases is a real problem. Long-term air leaks prolong postoperative hospitalization and result in an additional financial burden on the healthcare system. We focused our study on the evaluation of fibrin glue (Tissucol) as an effective means to minimize or prevent air leaks. METHODS: We initiated a case control study whereby 100 patients underwent precision resections for lung metastases. The subjects were divided into 2 groups, both with 50 patients: group 1 was treated with fibrin glue and group 2 with cauterization. Evaluation parameters consisted of the following: air leak duration, expected complications, drain time, and in-hospital stay. RESULTS: In group 1, air-leak time was 2.68 +/- 1.72 days, versus 7.80 +/- 8.52 for group 2 (P <.001). In group 1, there were 2% complications, whereas in group 2 there were 28% (P <.001). Drain time was 4.54 +/- 1.83 days for group 1 and 9.54 +/- 8.35 for group 2 (P <.001). In-hospital stay was 6.54 +/- 1.83 days for group 1 and 11.54 +/- 8.35 for group 2 (P <.001). CONCLUSIONS: In the group treated with fibrin glue, we observed significant advantages. Our experience shows that the use of fibrin glue can improve aerostasis control in nonanatomical resections with high risk of air leak. PMID- 12734096 TI - Renal cell carcinoma induces prostaglandin E2 and T-helper type 2 cytokine production in peripheral blood mononuclear cells. AB - BACKGROUND: Patients with renal cell carcinoma (RCC) do not develop an effective antitumor immune response, despite significant infiltration by lymphocytes. Tumor production of immunosuppressive factors may account for this failure. The object of this study was to investigate the production of immunosuppressive mediators, especially prostaglandin E(2) (PGE(2)), by RCC. METHODS: Peripheral blood mononuclear cells (PBMC) were cocultured with conditioned medium (CM) from human RCC cell lines in the presence or absence of NS-398, a selective cyclooxygenase 2 (COX-2) inhibitor. Supernatants were analyzed for levels of PGE(2), interleukin (IL)-10, IL-6, IL-2, interferon-gamma, and IL-12. The effects of RCC CM on PBMC proliferation were also examined. The expression of basal and stimulated COX-2 messenger RNA in the cell lines was assessed by reverse transcriptase-polymerase chain reaction. RESULTS: RCC CM significantly increased PGE(2) production by PBMC. T-helper type 2 (Th2) cytokine production was also significantly increased. Th1 cytokines were unchanged or decreased. RCC CM increased proliferation of PBMC. Coculture with NS-398 reduced PBMC PGE(2) production to below control levels and significantly decreased IL-6 production and PBMC proliferation. NS-398 had no effect on cellular production of IL-10 or Th1 cytokines. CONCLUSIONS: Human RCC inhibits the host antitumor immune response by promoting PGE(2) production and Th2 cytokines in PBMC. Selective inhibition of COX-2 may have a role in abrogating this effect. PMID- 12734097 TI - Thermal enhancement of new chemotherapeutic agents at moderate hyperthermia. AB - BACKGROUND: Hyperthermia enhances the cytotoxicity of some chemotherapeutic agents. We have studied the effect of moderate hyperthermia (41.5 degrees C) on the cytotoxicity of five new chemotherapeutic agents (docetaxel, paclitaxel, irinotecan, oxaliplatin, and gemcitabine) and melphalan against a spontaneous murine fibrosarcoma. METHODS: The tumor was an early-generation isotransplant of a spontaneous C3Hf/Sed mouse fibrosarcoma, FSa-II. Hyperthermia was administered by immersing the tumor-bearing foot into a constant temperature water bath set at 41.5 degrees C for 30 minutes when the tumor reached 34 mm(3). Chemotherapy was administered intraperitoneally immediately before hyperthermia. Tumor response was studied by the mean tumor growth time and the mean tumor growth delay time. RESULTS: Hyperthermia significantly increased the tumor growth times of the animals treated with docetaxel, irinotecan, and gemcitabine at low dose and these drugs plus oxaliplatin at high dose. Docetaxel at high dose showed the greatest control of tumor growth by hyperthermia, with a 26% reduction. Concerning the taxanes, paclitaxel cytotoxicity was not enhanced by hyperthermia, but docetaxel was enhanced by hyperthermia at both doses of drug. CONCLUSIONS: Moderate hyperthermia increases the cytotoxicity of docetaxel, irinotecan, and gemcitabine on mouse fibrosarcoma. Paclitaxel did not show heat enhancement. Oxaliplatin and docetaxel showed greater heat enhancement when the drug dose was high. PMID- 12734098 TI - Prostaglandin e(2) suppresses NK activity in vivo and promotes postoperative tumor metastasis in rats. AB - BACKGROUND: Prostaglandins (PGs) were shown in vitro to suppress several functions of cellular immunity. It is unclear, however, whether physiological levels of PGs can suppress cellular immunity in vivo and whether such suppression would compromise postoperative host resistance to metastasis. METHODS: Fischer 344 rats were administered PGE(2) in doses (18 to 300 micro g/kg subcutaneously) that increased the serum levels approximately 2- to 4-fold. We then assessed the number and activity of circulating natural killer (NK) cells, as well as rats' resistance to experimental metastasis of a syngeneic NK-sensitive tumor (MADB106). To study whether endogenously released PGs after surgery compromise these indices, we tested whether laparotomy adversely affects them and whether a cyclooxygenase-synthesis inhibitor, indomethacin (4 mg/kg), attenuates these effects. RESULTS: PGE(2) dose-dependently suppressed NK activity per NK cell and dose-dependently increased 4- and 24-hour MADB106 lung tumor retention (LTR); 240 micro g/kg of PGE(2) quadrupled the number of lung metastases counted 3 weeks later. Selective depletion of NK cells abrogated the promotion of LTR by PGE(2). Surgery significantly suppressed NK activity and increased MADB106 LTR, and indomethacin halved these effects without affecting nonoperated rats. CONCLUSIONS: PGE(2) is a potent in vivo suppressor of NK activity, and its postoperative release may promote tumor recurrence. PMID- 12734099 TI - Effect of isolated removal of either basolateral HCO-3 or basolateral CO2 on HCO 3 reabsorption by rabbit S2 proximal tubule. AB - The equilibrium CO2+H2O right arrow over left arrow H++HCO3- had made it impossible to determine how isolated changes in basolateral CO2 ([CO2]) or HCO3- concentration ([HCO3-]), at a fixed basolateral pH, modulate renal HCO3- or reabsorption. In the present study, we have begun to address this issue by measuring HCO3- reabsorption (JHCO3) and intracellular pH (pHi) in isolated perfused rabbit S2 proximal tubules exposed to three different basolateral (bath) solutions: 1) equilibrated 5% CO2/22 mM HCO3-/pH 7.40, 2) an out-of-equilibrium (OOE) solution containing 5% CO2/pH 7.40 but minimal HCO3- ("pure CO2"), and 3) an OOE solution containing 22 mM HCO3-/pH 7.40 but minimal CO2 ("pure HCO3-"). Tubule lumens were constantly perfused with equilibrated 5% CO2/22 mM HCO3-. Compared with the equilibrated bath solution (JHCO3 = 76.5 +/- 7.7 pmol.min-1.mm 1, pHi = 7.09 +/- 0.04), the pure CO2 bath solution increased JHCO3 by approximately 25% but decreased pHi by 0.19. In contrast, the pure HCO3- bath solution decreased JHCO3 by 37% but increased pHi by 0.24. Our data are consistent with two competing hypotheses: 1) the isolated removal of basolateral HCO3- (or CO2) causes a pHi decrease (increase) that in turn raises (lowers) JHCO3; and 2) HCO3- removal raises JHCO3 by reducing inhibition of basolateral Na/HCO3 cotransport and/or reducing HCO3- backleak, whereas CO2 removal lowers JHCO3 by reducing stimulation of a CO2 sensor. PMID- 12734100 TI - Determinants of renal afferent arteriolar actions of bradykinin: evidence that multiple pathways mediate responses attributed to EDHF. AB - The determinants of bradykinin (BK)-induced afferent arteriolar vasodilation were investigated in the in vitro perfused hydronephrotic rat kidney. BK elicited a concentration-dependent vasodilation of afferent arterioles that had been preconstricted with ANG II (0.1 nmol/l), but this dilation was transient in character. Pretreatment with the nitric oxide synthase inhibitor N(omega)-nitro-L arginine methyl ester (100 micromol/l) and the cyclooxygenase inhibitor ibuprofen (10 micromol/l) did not prevent this dilation when tone was established by ANG II but fully blocked the response when tone was established by elevated extracellular KCl, which suggests roles for both NO and endothelium-derived hyperpolarizing factor (EDHF). We had previously shown that the EDHF-like response of the afferent arteriole evoked by ACh was fully abolished by a combination of charybdotoxin (ChTX;10 nmol/l) and apamin (AP; 1 micromol/l). However, in the current study, treatment with ChTX plus AP only reduced the EDHF like component of the BK response from 98 +/- 5 to 53 +/- 6% dilation. Tetraethylammonium (TEA; 1 mmol/l), which had no effect on the EDHF-induced vasodilation associated with ACh, reduced the EDHF-like response to BK to 88 +/- 3% dilation. However, the combination of TEA plus ChTX plus AP abolished the response (0.3 +/- 1% dilation). Similarly, 17-octadecynoic acid (17-ODYA) did not prevent the dilation when it was administered alone (77 +/- 9% dilation) but fully abolished the EDHF-like response when added in combination with ChTX plus AP (-0.5 +/- 4% dilation). These findings suggest that BK acts via multiple EDHFs: one that is similar to that evoked by ACh in that it is blocked by ChTX plus AP, and a second that is blocked by either TEA or 17-ODYA. Our finding that a component of the BK response is sensitive to TEA and 17-ODYA is consistent with previous suggestions that the EDHF released by BK is an epoxyeicosatrienoic acid. PMID- 12734101 TI - Immortalized epithelial cells from human autosomal dominant polycystic kidney cysts. AB - Autosomal dominant polycystic kidney disease (ADPKD) is the result of mutations in one allele of the PKD1 or PKD2 genes, followed by "second hit" somatic mutations of the other allele in renal tubule cells. Continued proliferation of clonal cells originating from different nephron segments leads to cyst formation. In vitro studies of the mechanisms of cyst formation have been hampered by the scarcity of nephrectomy specimens and the limited life span of cyst-derived cells in primary culture. We describe the development of a series of immortalized epithelial cell lines from over 30 individual renal cysts obtained from 11 patients with ADPKD. The cells were immortalized with either wild-type (WT) or temperature-sensitive (TS) recombinant adeno-simian virus (SV)40 viruses. SV40 DNA integration into the cell genome was verified by PCR analysis. The cells have been passaged over 50 times with no apparent phenotypic change. By light microscopy, the cells appear pleomorphic but mostly polygonal and resemble the primary cultures. Transmission electron microscopy shows polarized epithelia with tight junctions. The SV40 large T antigen was detected by immunocytochemistry and by Western blot analysis at 37 degrees C in the WT cell lines and at 33 degrees C in the TS cell lines. It disappeared in TS cells 72 h following transfer to 39 degrees C. The majority (29) of the cell lines show binding of Dolichos biflorus lectin, suggesting distal tubule origin. Three cell lines show binding of Lotus tetragonolobus lectin or express aminopeptidase N, suggesting proximal tubule origin. Three cell lines were derived from a mixture of cysts and express features of both tubules. The PKD1 and PKD2 mRNA and protein were detected in all cells by RT-PCR and by immunocytochemistry. The majority of the cells tested also express the epidermal growth factor receptor, cystic fibrosis transmembrane conductance regulator, epithelial sodium channel, and renin. These new series of cyst-derived cell lines represent useful and readily available in vitro models for studying the cellular and molecular biology of ADPKD. PMID- 12734102 TI - Angiotensin receptor subtypes in thin and muscular juxtamedullary efferent arterioles of rat kidney. AB - ANG II controls the vascular tone of pre- and postglomerular arterioles, and thereby glomerular filtration, through binding to either AT1A, AT1B, or AT2 receptors. AT1 receptors, which are coupled to intracellular Ca2+ signaling, have vasoconstricting effects, whereas AT2 receptors, whose signaling mechanism is unknown, induce vasodilatation. The angiotensin receptors have been characterized in afferent arterioles, which express the three types of receptors, but not in efferent arterioles. Two subpopulations of juxtamedullary efferent arterioles, muscular ones which terminate as vasa rectae and thin ones which terminate as peritubular capillaries, have been described. They display functional heterogeneity with regard to the ANG II response. To evaluate whether these differences are associated with differential expression of ANG II receptors, we examined the expression pattern of AT1A, AT1B, and AT2 receptor mRNAs by RT-PCR in these arterioles and studied the effect of valsartan, a specific AT1-receptor antagonist. Results indicate that muscular arterioles express AT1A, AT1B, and AT2 receptors, whereas thin arterioles only express the AT1A and AT2 types, and at a much lower level. Valsartan fully inhibited ANG II-induced increases in intracellular Ca2+ in both arteriolar types, but with different kinetics. In muscular arterioles, inhibition was monoexponential, whereas it displayed a marked positive cooperativity in thin arterioles. Finally, the apparent affinity for valsartan was higher in muscular than in thin arterioles. In conclusion, this study further documents the differences between muscular and thin efferent arterioles with regard to ANG II signalization in the rat kidney. PMID- 12734103 TI - Claudins form ion-selective channels in the paracellular pathway. Focus on "Claudin extracellular domains determine paracellular charge selectively and resistance but not tight junction fibril architecture". PMID- 12734104 TI - Stretch-induced contractile differentiation of vascular smooth muscle: sensitivity to actin polymerization inhibitors. AB - Signaling mechanisms for stretch-dependent growth and differentiation of vascular smooth muscle were investigated in mechanically loaded rat portal veins in organ culture. Stretch-dependent protein synthesis was found to depend on endogenous release of angiotensin II. Autoradiography after [(35)S]methionine incorporation revealed stretch-dependent synthesis of several proteins, of which SM22 and actin were particularly prominent. Inhibition of RhoA activity by cell-permeant C3 toxin increased tissue mechanical compliance and reduced stretch-dependent extracellular signal-regulated kinase (ERK)1/2 activation, growth, and synthesis of actin and SM22, suggesting a role of the actin cytoskeleton. In contrast, inhibition of Rho-associated kinase by Y-27632 did not reduce ERK1/2 phosphorylation or actin and SM22 synthesis and did not affect tissue mechanical compliance but still inhibited overall growth. The actin polymerization inhibitors latrunculin B and cytochalasin D both inhibited growth and caused increased tissue compliance. Whereas latrunculin B concentration-dependently reduced actin and SM22 synthesis, cytochalasin D did so at low (10(-8) M) but not at high (10(-6) M) concentration. The results show that stretch stabilizes the contractile smooth muscle phenotype. Stretch-dependent differentiation marker expression requires an intact cytoskeleton for stretch sensing, control of protein expression via the level of unpolymerized G-actin, or both. PMID- 12734105 TI - Apoptosis repressor with caspase domain inhibits cardiomyocyte apoptosis by reducing K+ currents. AB - Cell shrinkage is an early prerequisite in programmed cell death, and cytoplasmic K(+) is a dominant cation that controls intracellular ion homeostasis and cell volume. Blockade of K(+) channels inhibits apoptotic cell shrinkage and attenuates apoptosis. We examined whether apoptotic repressor with caspase recruitment domain (ARC), an antiapoptotic protein, inhibits cardiomyocyte apoptosis by reducing K(+) efflux through voltage-gated K(+) (Kv) channels. In heart-derived H9c2 cells, whole cell Kv currents (I(K(V))) were isolated by using Ca(2+)-free extracellular (bath) solution and including 5 mM ATP and 10 mM EGTA in the intracellular (pipette) solution. Extracellular application of 5 mM 4 aminopyridine (4-AP), a blocker of Kv channels, reversibly reduced I(K(V)) by 50 60% in H9c2 cells. The remaining currents during 4-AP treatment may be generated by K(+) efflux through 4-AP-insensitive K(+) channels. Overexpression of ARC in heart-derived H9c2 cells significantly decreased I(K(V)), whereas treatment with staurosporine, a potent apoptosis inducer, enhanced I(K(V)) in wild-type cells. The staurosporine-induced increase in I(K(V)) was significantly suppressed and the staurosporine-mediated apoptosis was markedly inhibited in cells overexpressing ARC compared with cells transfected with the control neomycin vector. These results suggest that the antiapoptotic effect of ARC is, in part, due to inhibition of Kv channels in cardiomyocytes. PMID- 12734106 TI - MHC-based fiber type and E-C coupling characteristics in mechanically skinned muscle fibers of the rat. AB - In this study, we investigated whether the previously established differences between fast- and slow-twitch single skeletal muscle fibers of the rat, in terms of myosin heavy chain (MHC) isoform composition and contractile function, are also detectable in excitation-contraction (E-C) coupling. We compared the contractile responsiveness of electrophoretically typed, mechanically skinned single fibers from the soleus (Sol), the extensor digitorum longus (EDL), and the white region of the sternomastoid (SM) muscle to t-system depolarization-induced activation. The quantitative parameters assessed were the amplitude of the maximum depolarization-induced force response (DIFR(max); normalized to the maximum Ca(2+)-activated force in that fiber) and the number of responses elicited until the force declined by 75% of DIFR(max) (R-D(75%)). The mean DIFR(max) values for type IIB EDL and type IIB SM fibers were not statistically different, and both were greater than the mean DIFR(max) for type I Sol fibers. The mean R-D(75%) for type IIB EDL fibers was greater than that for type I Sol fibers as well as type IIB SM fibers. These data suggest that E-C coupling characteristics of mechanically skinned rat single muscle fibers are related to MHC-based fiber type and the muscle of origin. PMID- 12734107 TI - DMT1, a physiologically relevant apical Cu1+ transporter of intestinal cells. AB - Despite important advances in the understanding of copper secretion and excretion, the molecular components of intestinal copper absorption remain a mystery. DMT1, also known as Nramp2 and DCT1, is the transporter responsible for intestinal iron uptake. Electrophysiological evidence suggests that DMT1 can also be a copper transporter. Thus we examined the potential role of DMT1 as a copper transporter in intestinal Caco-2 cells. Treatment of cells with a DMT1 antisense oligonucleotide resulted in 80 and 48% inhibition of iron and copper uptake, respectively. Cells incorporated considerable amounts of copper as Cu(1+), whereas Cu(2+) transport was about 10-fold lower. Cu(1+) inhibited apical Fe(2+) transport. Fe(2+), but not Fe(3+), effectively inhibited Cu(1+) uptake. The iron content of the cells influenced both copper and iron uptake. Cells with low iron content transported fourfold more iron and threefold more copper than cells with high iron content. These results demonstrate that DMT1 is a physiologically relevant Cu(1+) transporter in intestinal cells, indicating that intestinal absorption of copper and iron are intertwined. PMID- 12734108 TI - Regulation of endothelial nitric oxide synthase by the actin cytoskeleton. AB - In the present study, the association of endothelial nitric oxide synthase (eNOS) with the actin cytoskeleton in pulmonary artery endothelial cells (PAEC) was examined. We found that the protein contents of eNOS, actin, and caveolin-1 were significantly higher in the caveolar fraction of plasma membranes than in the noncaveolar fraction of plasma membranes in PAEC. Immunoprecipitation of eNOS from lysates of caveolar fractions of plasma membranes in PAEC resulted in the coprecipitation of actin, and immunoprecipitation of actin from lysates of caveolar fractions resulted in the coprecipitation of eNOS. Confocal microscopy of PAEC, in which eNOS was labeled with fluorescein, F-actin was labeled with Texas red-phalloidin, and G-actin was labeled with deoxyribonuclease I conjugated with Texas red, also demonstrated an association between eNOS and F-actin or G actin. Incubation of purified eNOS with purified F-actin and G-actin resulted in an increase in eNOS activity. The increase in eNOS activity caused by G-actin was much higher than that caused by F-actin. Incubation of PAEC with swinholide A, an actin filament disruptor, resulted in an increase in eNOS activity, eNOS protein content, and association of eNOS with G-actin and in a decrease in the association of eNOS with F-actin. The increase in eNOS activity was higher than that in eNOS protein content in swinholide A-treated cells. In contrast, exposure of PAEC to phalloidin, an actin filament stabilizer, caused decreases in eNOS activity and association of eNOS with G-actin and increases in association of eNOS with F-actin. These results suggest that eNOS is associated with actin in PAEC and that actin and its polymerization state play an important role in the regulation of eNOS activity. PMID- 12734109 TI - Molecular cloning of NHE1 from winter flounder RBCs: activation by osmotic shrinkage, cAMP, and calyculin A. AB - In this report, we describe the cloning, cellular localization, and functional characteristics of Na(+)/H(+) exchanger 1 (NHE1) from red blood cells of the winter flounder Pseudopleuronectes americanus (paNHE1). The paNHE1 protein localizes primarily to the marginal band and exhibits a 74% similarity to the trout beta-NHE, and 65% to the human NHE1 (hNHE1). Functionally, paNHE1 shares characteristics of both beta-NHE and hNHE1 in that it is activated both by manipulations that increase cAMP and by cell shrinkage, respectively. In accordance, the paNHE1 protein exhibits both protein kinase A consensus sites as in beta-NHE and a region of high homology to that required for shrinkage dependent activation of hNHE1. After shrinkage-dependent activation of paNHE1 and resulting activation of a Cl(-)/HCO(3)(-) exchanger, their parallel operation results in net uptake of NaCl and osmotically obliged water. Activation of paNHE1 by cAMP is at least additive to that elicited by osmotic shrinkage, suggesting that these stimuli regulate paNHE1 by distinct mechanisms. Finally, exposure to the serine/threonine phosphatase inhibitor calyculin A potently activates paNHE1, and this activation is also additive to that induced by shrinkage or cAMP. PMID- 12734110 TI - Effect of interleukin-1beta and tumor necrosis factor-alpha on gene expression in human endothelial cells. AB - Interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) are two major cytokines that rise to relatively high levels during systemic inflammation, and the endothelial cell (EC) response to these cytokines may explain some of the dysfunction that occurs. To better understand the cytokine-induced responses of EC at the gene expression level, human umbilical vein EC were exposed to IL-1beta or TNF-alpha for various times and subjected to cDNA microarray analyses to study alterations in their mRNA expression. Of approximately 4,000 genes on the microarray, expression levels of 33 and 58 genes appeared to be affected by treatment with IL-1beta and TNF-alpha, respectively; 25 of these genes responded to both treatments. These results suggest that the effects of IL-1beta and TNF alpha on EC are redundant and that it may be necessary to suppress both cytokines simultaneously to ameliorate the systemic response. PMID- 12734111 TI - Chloro(2,2':6',2"-terpyridine) platinum inhibition of the renal Na+,K+-ATPase. AB - Chloro(2,2':6',2"-terpyridine) platinum, a bulky, hydrophilic reagent, inhibited the renal sodium pump with a single exponential time course. K(+) increased the rate constant of the reaction by about twofold; the K(+) concentration dependence was monotonic, with a half-maximal effect observed at 1 mM, consistent with K(+) acting at a transport site. Na(+), Mg(2+), eosin, and vanadate did not significantly alter the rate of reaction. The results of proteolysis and mass spectrometer analysis were consistent with terpyridine platinum labeling of Cys452, Cys456, or Cys457. Because phenylarsine oxide reacts with vicinal cysteines and did not prevent terpyridine platinum modification, terpyridine platinum most likely modifies Cys452. This modification prevents ADP binding; interestingly, the analogous residue in sarco(endo)plasmic reticulum Ca(2+) ATPase (SERCA) is on the exterior of the nucleotide-binding pocket. Thus it appears that the terpyridine platinum residue is more accessible in the presence of K(+) than in its absence and that terpyridine platinum modification prevents nucleotide binding. PMID- 12734112 TI - Cyclosporin A-induced hair growth in mice is associated with inhibition of calcineurin-dependent activation of NFAT in follicular keratinocytes. AB - One of the most common side effects of treatment with cyclosporin A (CsA) is hypertrichosis. This study shows that calcineurin activity is associated with hair keratinocyte differentiation in vivo, affecting nuclear factor of activated T cells (NFAT1) activity in these cells. Treatment of nude or C57BL/6 depilated normal mice with CsA inhibited the expression of keratinocyte terminal differentiation markers associated with catagen, along with the inhibition of calcineurin and NFAT1 nuclear translocation. This was associated with induction of hair growth in nude mice and retardation of spontaneous catagen induction in depilated normal mice. Furthermore, calcineurin inhibition blocked the expression of p21(waf/cip1) and p27(kip1), which are usually induced with differentiation. This was also associated with an increase in interleukin-1alpha expression (nude mice), a decrease in transforming growth factor-beta (nude and normal mice), and no change in keratinocyte growth factor expression in the skin. Retardation of catagen in CsA-treated mice was accompanied by significant alterations in apoptosis-related gene product expression in hair follicle keratinocytes. The ratio of the anti-apoptotic Bcl-2 to proapoptotic Bax expression increased, and expression of p53 and interleukin-1beta converting enzyme activity decreased. These data provide the first evidence that calcineurin is functionally active in follicular keratinocytes and that inhibition of the calcineurin-NFAT1 pathway in these cells in vivo by CsA enhances hair growth. PMID- 12734113 TI - Structural elements of kallistatin required for inhibition of angiogenesis. AB - Kallistatin is a serpin first identified as a specific inhibitor of tissue kallikrein. Our recent studies showed that kallikrein promoted angiogenesis, whereas kallistatin inhibited angiogenesis and tumor growth. This study is aimed to identify the structural elements of kallistatin essential for its antiangiogenic function. Kallistatin mutants at the hinge region (A377T) and a major heparin-binding domain (K312A/K313A) were created by site-directed mutagenesis. Recombinant kallistatin mutant A377T did not bind or inhibit tissue kallikrein activity. Wild-type kallistatin and kallistatin mutant A377T, but not kallistatin mutant K312A/K313A lacking heparin-binding activity, inhibited VEGF induced proliferation, growth, and migration of human microvascular endothelial cells. Similarly, wild-type kallistatin and kallistatin mutant A337T, but not kallistatin mutant K312A/K313A, significantly inhibited VEGF-induced capillary tube formation of cultured endothelial cells in Matrigel and capillary formation in Matrigel implants in mice. To elucidate the role of the heparin-binding domain in modulating angiogenesis, we showed that wild-type kallistatin interrupted the binding of (125)I-labeled VEGF to endothelial cells, whereas kallistatin mutant K312A/K313A did not interfere with VEGF binding. Consequently, wild-type kallistatin, but not kallistatin mutant K312A/K313A, suppressed VEGF-induced phosphorylation of Akt. Taken together, these results indicate that the heparin binding domain, but not the reactive site loop of kallistatin, is essential for inhibiting VEGF-induced angiogenesis. PMID- 12734114 TI - PPARgamma coactivator-1alpha expression during thyroid hormone- and contractile activity-induced mitochondrial adaptations. AB - The transcriptional coactivator the peroxisome proliferator-activated receptor gamma coactivator-1alpha (PGC-1alpha) has been identified as an important mediator of mitochondrial biogenesis based on its ability to interact with transcription factors that activate nuclear genes encoding mitochondrial proteins. The induction of PGC-1alpha protein expression under conditions that provoke mitochondrial biogenesis, such as contractile activity or thyroid hormone (T(3)) treatment, is not fully characterized. Thus we related PGC-1alpha protein expression to cytochrome c oxidase (COX) activity in 1) tissues of varying oxidative capacities, 2) tissues from animals treated with T(3), and 3) skeletal muscle subject to contractile activity both in cell culture and in vivo. Our results demonstrate a strong positive correlation (r = 0.74; P < 0.05) between changes in PGC-1alpha and COX activity, used as an index of mitochondrial adaptations. The highest constitutive levels of PGC-1alpha were found in the heart, whereas the lowest were measured in fast-twitch white muscle and liver. T(3) increased PGC-1alpha content similarly in both fast- and slow-twitch muscle, as well as in the liver, but not in heart. T(3) also induced early (6 h) increases in AMP-activated protein kinase (AMPKalpha) activity, as well as later (5 day) increases in p38 MAP kinase activity in slow-twitch, but not in fast twitch, muscle. Contractile activity provoked early increases in PGC-1alpha, coincident with increases in mitochondrial transcription factor A (Tfam), and nuclear respiratory factor-1 (NRF-1) protein expression, suggesting that PGC 1alpha is physiologically important in coordinating the expression of the nuclear and mitochondrial genomes. Ca(2+) ionophore treatment of muscle cells led to an approximately threefold increase in PGC-1alpha protein, and contractile activity induced rapid and marked increases in both p38 MAP kinase and AMPKalpha activities. 5-Aminoimidazole-4-carboxamide-1-beta-d-ribofuranoside (AICAR) treatment of muscle cells also led to parallel increases in AMPKalpha activity and PGC-1alpha protein levels. These data are consistent with observations that indicate that increases in PGC-1alpha protein are affected by Ca(2+) signaling mechanisms, AMPKalpha activity, as well as posttranslational phosphorylation events that increase PGC-1alpha protein stability. Our data support a role for PGC-1alpha in the physiological regulation of mitochondrial content in a variety of tissues and suggest that increases in PGC-1alpha expression form part of a unifying pathway that promotes both T(3)- and contractile activity-induced mitochondrial adaptations. PMID- 12734115 TI - Muscle force emerges from dynamic actin-myosin networks, not from independent force generators. PMID- 12734116 TI - A piece of my mind. Time to myself. PMID- 12734117 TI - Researchers probe aortic stenosis: an active, potentially treatable disease process? PMID- 12734118 TI - Experts weigh prevention, therapy for ocular vaccinia in smallpox vaccinees. PMID- 12734119 TI - Insomniac artists explore night's terrain. PMID- 12734120 TI - From the Centers for Disease Control and Prevention. Severe Acute Respiratory Syndrome (SARS) and coronavirus testing--United States, 2003. PMID- 12734124 TI - Cesarean delivery and risk of herpes simplex virus infection. PMID- 12734123 TI - Cesarean delivery and risk of herpes simplex virus infection. PMID- 12734125 TI - Management of pressure ulcers. PMID- 12734126 TI - Long-term cognitive function in very low-birth-weight infants. PMID- 12734127 TI - Management of pressure ulcers. PMID- 12734128 TI - Management of pressure ulcers. PMID- 12734129 TI - Management of pressure ulcers. PMID- 12734130 TI - Intervening in abusive relationships. PMID- 12734131 TI - Listing contributions of investigators in research groups. PMID- 12734132 TI - Impact of electron beam tomography, with or without case management, on motivation, behavioral change, and cardiovascular risk profile: a randomized controlled trial. AB - CONTEXT: Although the use of electron beam tomography (EBT) as a motivational tool to change behavior is practiced, its efficacy has not been studied. OBJECTIVE: To assess the effects of incorporating EBT as a motivational factor into a cardiovascular screening program in the context of either intensive case management (ICM) or usual care by assessing its impact over 1 year on a composite measure of projected risk. DESIGN: Randomized controlled trial with a 2 x 2 factorial design and 1 year of follow-up. SETTING AND PARTICIPANTS: A consecutive sample of 450 asymptomatic active-duty US Army personnel aged 39 to 45 years stationed within the Washington, DC, area and scheduled to undergo a periodic Army-mandated physical examination were enrolled between January 1999 and March 2001 (mean age, 42 years; 79% male; 66 [15%] had coronary calcification; mean [SD] predicted 10-year coronary risk, 5.85% [3.85%]). INTERVENTIONS: Patients were randomly assigned to 1 of 4 intervention arms: EBT results provided in the setting of either ICM (n = 111) or usual care (n = 119) or EBT results withheld in the setting of either ICM (n = 124) or usual care (n = 96). MAIN OUTCOME MEASURE: The primary outcome measure was change in a composite measure of risk, the 10-year Framingham Risk Score (FRS). RESULTS: Comparing the groups who received EBT results with those who did not, the mean absolute risk change in 10 year FRS was +0.30 vs +0.36 (P =.81). Comparing the groups who received ICM with those who received usual care, the mean absolute risk change in 10-year FRS was 0.06 vs +0.74 (P =.003). Improvement or stabilization of cardiovascular risk was noted in 157 patients (40.2%). In multivariable analyses predicting change in FRS, after controlling for knowledge of coronary calcification, motivation for change, and multiple psychological variables, only the number of risk factors (odds ratio, 1.42; 95% confidence interval, 1.16-1.75 for each additional risk factor) and receipt of ICM (odds ratio, 1.62; 95% confidence interval, 1.04-2.52) were associated with improved or stabilized projected risk. CONCLUSIONS: Using coronary calcification screening to motivate patients to make evidence-based changes in risk factors was not associated with improvement in modifiable cardiovascular risk at 1 year. Case management was superior to usual care in the management of risk factors. PMID- 12734133 TI - Pacemaker therapy for prevention of syncope in patients with recurrent severe vasovagal syncope: Second Vasovagal Pacemaker Study (VPS II): a randomized trial. AB - CONTEXT: Three previous small randomized trials have reported that pacemaker therapy is beneficial for patients with severe recurrent vasovagal syncope. However, because these trials were not double blind, they may have been biased in their assessment of outcomes and had a placebo effect of surgery. OBJECTIVE: To determine if pacing therapy reduces the risk of syncope in patients with vasovagal syncope. DESIGN, SETTING, AND PATIENTS: A randomized double-blind trial of pacemaker therapy in outpatients referred to syncope specialists at 15 centers from September 1998 to April 2002. In the year prior to randomization, patients had had a median of 4 episodes of syncope. Patients were followed up for up to 6 months. INTERVENTION: After implantation of a dual chamber pacemaker, 100 patients were randomly assigned to receive dual-chamber pacing (DDD) with rate drop response or to have only sensing without pacing (ODO). MAIN OUTCOME MEASURE: Time to first recurrence of syncope. RESULTS: No patients were lost to follow-up. Of the 52 patients randomized to ODO, 22 (42%) had recurrent syncope within 6 months compared with 16 (33%) of 48 patients in the DDD group. The cumulative risk of syncope at 6 months was 40% (95% confidence interval [CI], 25%-52%) for the ODO group and 31% (95% CI, 17%-43%) for the DDD group. The relative risk reduction in time to syncope with DDD pacing was 30% (95% CI, -33% to 63%; 1 sided P =.14). Lead dislodgement or repositioning occurred in 7 patients. One patient had vein thrombosis, another had pericardial tamponade leading to removal of the pacemaker system, and a third had infection involving the pacemaker generator. CONCLUSIONS: In this double-blind randomized trial, pacing therapy did not reduce the risk of recurrent syncope in patients with vasovagal syncope. Because of the weak evidence of efficacy of pacemaker therapy and the risk of complications, pacemaker therapy should not be recommended as first-line therapy for patients with recurrent vasovagal syncope. PMID- 12734134 TI - Incidence of sleep-disordered breathing in an urban adult population: the relative importance of risk factors in the development of sleep-disordered breathing. AB - CONTEXT: Sleep-disordered breathing (SDB) is both prevalent and associated with serious chronic illness. The incidence of SDB and the effect of risk factors on this incidence are unknown. OBJECTIVE: To determine the 5-year incidence of SDB overall and as influenced by risk factors. DESIGN, SETTING, AND PARTICIPANTS: Of the 1149 participants in the Cleveland Family Study, those aged 18 years or older, from either case or control families, who had 2 in-home sleep studies 5 years apart. The first had to have been performed before June 30, 1997, and had to have normal results (apnea hypopnea index [AHI] <5). Data included questionnaire information on medical and family history, SDB symptoms; measurement of height, weight, blood pressure, waist and hip circumference, and serum cholesterol concentration; and overnight sleep monitoring. MAIN OUTCOME MEASURE: Apnea hypopnea index, defined as number of apneas and hypopneas per hour of sleep. Sleep-disordered breathing was defined by an AHI of at least 10 (mild to moderate) or of at least 15 (moderate). RESULTS: Forty-seven (16%) of 286 eligible participants, (95% confidence interval [CI], 13%-21%) had a second-study AHI of at least 10 and 29 (10%) participants (95% CI, 7%-14%) had a second-study AHI result of at least 15. For the AHI results of at least 15, we estimate that about 2.5% may represent test variability. By ordinal logistic regression analysis, AHI was significantly associated with age (odds ratio [OR] per 10-year increase, 1.79; 95% CI, 1.41-2.27), body mass index (BMI; OR per 1-unit increase, 1.14; 95% CI, 1.10-1.19), sex (OR for men vs women, 4.12; 95% CI, 2.29-7.43), waist-hip ratio (OR per 0.1 unit increase, 1.61; 95% CI, 1.04-2.28), and serum cholesterol concentration (OR per 10-mg/dL [0.25-mmol/L] increase, 1.11; 95% CI, 1.03-1.19). Interactions were noted between age and both sex (P =.003) and BMI (P =.05). The OR for increased AHI per 10-year age increase was 2.41 in women (95% CI, 1.78-3.26) and 1.15 in men (95% CI, 0.78-1.68), with the male vs female OR decreasing from 5.04 (95% CI, 2.19-11.6) at age 30 years to 0.54 (95% CI, 0.15 1.99) at age 60 years. The OR for increased AHI per 1-unit increase in BMI decreased from 1.21 (95% CI, 1.11-1.31) at age 20 years to 1.05 (95% CI, 0.96 1.15) at age 60 years. CONCLUSIONS: The 5-year incidence is about 7.5% for moderately severe SDB and 16% (or less) for mild to moderately severe SDB. Incidence of SDB is influenced independently by age, sex, BMI, waist-hip ratio, and serum cholesterol concentration. Predominance in men diminishes with increasing age, and by age 50 years, incidence rates among men and women are similar. The effect of BMI also decreases with age and may be negligible at age 60 years. PMID- 12734135 TI - Hospice use among Medicare managed care and fee-for-service patients dying with cancer. AB - CONTEXT: For most patients aged 65 years or older with cancer, hospice services are uniformly covered by Medicare. Hospice care is believed to improve care for patients at the end of life. However, few patients use hospice and others enroll too late to maximize the benefits of hospice services. OBJECTIVES: Because type of insurance may affect use, we examined whether patients with Medicare managed care insurance enrolled in hospice earlier and had longer hospice stays than patients with Medicare fee-for-service (FFS) insurance. DESIGN AND SETTING: Retrospective analysis of the last year of life using the Linked Medicare-Tumor Registry Database in 1 of 9 Surveillance, Epidemiology, and End Results program coverage areas. PATIENTS: A total of 260 090 Medicare beneficiaries aged 66 years or older diagnosed with first primary lung (n = 62 117), colorectal (n = 57 260), prostate (n = 59 826), female breast (n = 37 609), bladder (n = 19 598), pancreatic (n = 11 378), gastric (n = 9599), or liver (n = 2703) cancer between January 1, 1973, and December 31, 1996, and who died between January 1, 1988, and December 31, 1998. MAIN OUTCOME MEASURES: Time from diagnosis to hospice entry and hospice length of stay for patients enrolled in FFS vs managed care plans after adjusting for patient demographics, tumor registry, year of hospice entry, and type and cancer stage. RESULTS: Of the 260 090 patients, most were men (59%), white (85%), and enrolled in FFS (89.7%). Only 54 937 patients (21.1%) received hospice care before death. Hospice use varied by type of primary cancer ranging from 31.8% of patients with pancreatic cancer to 15.6% with bladder cancer. Managed care patients were more likely to use hospice than FFS patients (32.4% vs 19.8%, P<.001). Among hospice patients, median (interquartile range) length of stay was longer for managed care vs FFS patients (32 days [11-82] vs 25 days [9 66], P<.001). After adjustment, managed care patients had higher rates of hospice enrollment (adjusted hazard ratio [HR], 1.38; 95% CI, 1.35-1.42) and had a longer length of stay (adjusted HR, 0.91; 95% CI, 0.88-0.94) vs FFS patients. Managed care patients were less likely to enroll in hospice within 7 days of their death (18.6% vs 22.6%, P<.001) and somewhat more likely to enroll in hospice more than 180 days before death (7.8% vs 6.1%, P<.001); the results for each of the 8 cancer diagnoses were similar. Hospice enrollment and length of stay among managed care vs FFS patients differed significantly by region. CONCLUSION: Medicare beneficiaries enrolled in managed care had consistently higher rates of hospice use and significantly longer hospice stays than those enrolled in FFS. Although these differences may reflect patient and family preferences, our findings raise the possibility that some managed care plans are more successful at facilitating or encouraging hospice use for patients dying with cancer. PMID- 12734136 TI - Effect of a national disaster on blood supply and safety: the September 11 experience. AB - CONTEXT: An understanding of characteristics of blood donors donating in times of crisis may help predict blood supply safety and donor return patterns. OBJECTIVES: To characterize the volume of donations and prevalence of infectious disease markers in blood donated by US donors responding to the September 11, 2001, terrorist attacks, and to evaluate return rates in those who donated for the first time. DESIGN: Cross-sectional survey data from the National Heart, Lung, and Blood Institute Retrovirus Epidemiology Donor Study for 4 weeks before and 4 weeks starting with September 11, 2001, and the corresponding 8-week period in 2000. SETTING AND PARTICIPANTS: A total of 327065 volunteer blood donors making 373628 allogeneic donations at 5 large regional US blood centers. MAIN OUTCOME MEASURES: Changes in number of donations overall and by first-time and repeat status, prevalence of infectious disease markers, estimated risks of transfusion-transmitted viral infections, and first-time donor return rates. RESULTS: About 20000 allogeneic donations were collected weekly in the 4 weeks preceding September 11, whereas approximately 49 000 (2.5-fold increase) and approximately 26000 to 28000 (1.3-fold to 1.4-fold increases) donations were made per week in the first and in the second through fourth weeks starting with September 11, respectively. All demographic groups donated more than usual after the attacks, and after adjusting for seasonal and annual variation there was a 5.2-fold (95% confidence interval, 5.0-5.4) increase in the number of first-time donations vs a 1.5-fold (1.4-1.5) increase in the number of repeat donations made in the first week starting on September 11 vs the 4 weeks before. The weekly proportion of repeat donors returning after not donating for 10 or more years increased from 2% before September 11 to 6% in the first week starting with September 11. Donations confirmed positive for human immunodeficiency virus (HIV), hepatitis C virus (HCV), and hepatitis B surface antigen nearly tripled between 1 week before September 11 (0.1%) and 1 week after the attacks (0.3%), largely explained by the increase in first-time and lapsed repeat donors. Estimated viral residual risks increased slightly after the attacks (HIV, 1/1.5 million vs 1/1.8 million donations; HCV, 1/1.3 million vs 1/1.6 million; hepatitis B virus, 1/140000 vs 1/170000). First-time donor 12-month return rates for 2000 and 2001 were similar, approximately 28% (P =.37) for donors in the first week starting with September 11 (or September 12, 2000) and 30% (P =.69) for the second to fourth weeks. CONCLUSIONS: The September 11 events resulted in an influx of first-time donors without substantial increase in absolute risk of transfusion-transmissible viral infections. First-time donor return rates were equally relatively low before and after the attacks, suggesting that those donating in times of crisis have return behaviors similar to those of other first time donors. Their relatively low return rates reinforce the need for education about the importance of donating regularly. PMID- 12734137 TI - Outpatient insulin therapy in type 1 and type 2 diabetes mellitus: scientific review. AB - CONTEXT: Newer insulin therapies, including the concept of physiologic basal prandial insulin and the availability of insulin analogues, are changing clinical diabetes care. The key to effective insulin therapy is an understanding of principles that, when implemented, can result in improved diabetes control. OBJECTIVE: To systematically review the literature regarding insulin use in patients with type 1 and type 2 diabetes mellitus (DM). DATA SOURCES: A MEDLINE search was performed to identify all English-language articles of randomized controlled trials involving insulin use in adults with type 1 or type 2 DM from January 1, 1980, to January 8, 2003. Bibliographies and experts were used to identify additional studies. STUDY SELECTION AND DATA EXTRACTION: Studies were included (199 for type 1 DM and 144 for type 2 DM, and 38 from other sources) if they involved human insulins or insulin analogues, were at least 4 weeks long with at least 10 patients in each group, and glycemic control and hypoglycemia were reported. Studies of insulin-oral combination were similarly selected. DATA SYNTHESIS: Twenty-eight studies for type 1 DM, 18 for type 2 DM, and 48 for insulin-oral combination met the selection criteria. In patients with type 1 DM, physiologic replacement, with bedtime basal insulin and a mealtime rapid-acting insulin analogue, results in fewer episodes of hypoglycemia than conventional regimens. Rapid-acting insulin analogues are preferred over regular insulin in patients with type 1 DM since they improve HbA1C and reduce episodes of hypoglycemia. In patients with type 2 DM, adding bedtime neutral protamine Hagedorn (isophane) insulin to oral therapy significantly improves glycemic control, especially when started early in the course of disease. Bedtime use of insulin glargine results in fewer episodes of nighttime hypoglycemia than neutral protamine Hagedorn regimens. For patients with more severe insulin deficiency, a physiologic insulin regimen should allow lower glycemic targets in the majority of patients. Adverse events associated with insulin therapy include hypoglycemia, weight gain, and worsening diabetic retinopathy if hemoglobin A1C levels decrease rapidly. CONCLUSIONS: Many options for insulin therapy are now available. Physiologic insulin therapy with insulin analogues is now relatively simple to use and is associated with fewer episodes of hypoglycemia. PMID- 12734138 TI - Using new insulin strategies in the outpatient treatment of diabetes: clinical applications. AB - Understanding when to use insulin and how to apply the principles of physiologic insulin replacement using existing and new insulins is a key step to improving diabetes care. Insulin analogues and premixed insulins increase physicians' and patients' ability to lower hemoglobin A1C levels with fewer episodes of hypoglycemia. Earlier use of insulin and more aggressive dose escalation are important steps in achieving treatment goals. This article discusses using bedtime insulin with oral agents, basal-prandial insulin strategies, and the new insulin analogues. PMID- 12734139 TI - Improving risk of coronary heart disease: can a picture make the difference? PMID- 12734140 TI - Is there an effective treatment for neurally mediated syncope? PMID- 12734141 TI - MSJAMA. Physician-assisted suicide. PMID- 12734142 TI - MSJAMA. Legal status of physician-assisted suicide. PMID- 12734144 TI - MSJAMA. The legal and political future of physician-assisted suicide. PMID- 12734143 TI - MSJAMA. Moral and practical challenges of physician-assisted suicide. PMID- 12734145 TI - JAMA patient page. Insulin. PMID- 12734148 TI - Neuromuscular blocking drugs and allergic risk. PMID- 12734147 TI - Clinical features and short-term outcomes of 144 patients with SARS in the greater Toronto area. AB - CONTEXT: Severe acute respiratory syndrome (SARS) is an emerging infectious disease that first manifested in humans in China in November 2002 and has subsequently spread worldwide. OBJECTIVES: To describe the clinical characteristics and short-term outcomes of SARS in the first large group of patients in North America; to describe how these patients were treated and the variables associated with poor outcome. DESIGN, SETTING, AND PATIENTS: Retrospective case series involving 144 adult patients admitted to 10 academic and community hospitals in the greater Toronto, Ontario, area between March 7 and April 10, 2003, with a diagnosis of suspected or probable SARS. Patients were included if they had fever, a known exposure to SARS, and respiratory symptoms or infiltrates observed on chest radiograph. Patients were excluded if an alternative diagnosis was determined. MAIN OUTCOME MEASURES: Location of exposure to SARS; features of the history, physical examination, and laboratory tests at admission to the hospital; and 21-day outcomes such as death or intensive care unit (ICU) admission with or without mechanical ventilation. RESULTS: Of the 144 patients, 111 (77%) were exposed to SARS in the hospital setting. Features of the clinical examination most commonly found in these patients at admission were self reported fever (99%), documented elevated temperature (85%), nonproductive cough (69%), myalgia (49%), and dyspnea (42%). Common laboratory features included elevated lactate dehydrogenase (87%), hypocalcemia (60%), and lymphopenia (54%). Only 2% of patients had rhinorrhea. A total of 126 patients (88%) were treated with ribavirin, although its use was associated with significant toxicity, including hemolysis (in 76%) and decrease in hemoglobin of 2 g/dL (in 49%). Twenty-nine patients (20%) were admitted to the ICU with or without mechanical ventilation, and 8 patients died (21-day mortality, 6.5%; 95% confidence interval [CI], 1.9%-11.8%). Multivariable analysis showed that the presence of diabetes (relative risk [RR], 3.1; 95% CI, 1.4-7.2; P =.01) or other comorbid conditions (RR, 2.5; 95% CI, 1.1-5.8; P =.03) were independently associated with poor outcome (death, ICU admission, or mechanical ventilation). CONCLUSIONS: The majority of cases in the SARS outbreak in the greater Toronto area were related to hospital exposure. In the event that contact history becomes unreliable, several features of the clinical presentation will be useful in raising the suspicion of SARS. Although SARS is associated with significant morbidity and mortality, especially in patients with diabetes or other comorbid conditions, the vast majority (93.5%) of patients in our cohort survived. PMID- 12734149 TI - The law and the physician as principal investigator in sponsored clinical trials. PMID- 12734146 TI - Severe acute respiratory syndrome: providing care in the face of uncertainty. PMID- 12734150 TI - Intravenous fluid loading with or without supplementary dextrose does not prevent nausea, vomiting and pain after laparoscopy. AB - PURPOSE: To examine the effects of iv compound sodium lactate (CSL) with and without caloric supplementation with dextrose on nausea, vomiting and pain following general anesthesia for laparoscopy. METHODS: We compared iv fluid loading with and without supplementary dextrose for the prevention of postoperative nausea and vomiting (PONV). In a prospective double-blinded controlled trial, 120 ASA I female patients undergoing elective gynecological laparoscopy were randomized to one of three groups, and received either: (a) CSL 1.5 mL.kg(-1) per hour fasting duration; (b) CSL, 1.5 mL.kg(-1) per hour fasting duration with 0.5 g.kg(-1) dextrose added in 50% formulation (CSL/dextrose); or (c) no iv fluid (control). RESULTS: Compared with control the percentage of patients who had no PONV within 24 hr of anesthesia in the CSL and CSL/dextrose groups was 78% vs 83% and 71%, P = 0.81 and P = 0.683 respectively. The numbers needed-to-harm for causing PONV episodes in CSL/dextrose vs CSL or control groups were 5.7 [95% confidence interval (CI), 5.57-5.91] and 8.2 (95% CI, 8.01-8.37) respectively. The number needed-to-treat for prevention of PONV episodes in CSL vs control was 19.2 (95% CI, 19.08-19.37). A greater proportion of patients in the CSL/dextrose group required narcotic analgesia in the postanesthetic care unit compared to those in the control group (16/35 vs 7/37, P = 0.03). The CSL/dextrose group also demonstrated hyperglycemia (serum glucose 14.0 +/- 3.94 vs 5.0 +/- 1.01 vs 5.2 +/- 0.9 mmol.L(-1), P < 0.0001) in the postanesthetic care unit compared to the CSL and control groups. The CSL/dextrose group also reported increased thirst at 24 hr compared to control (20/35 vs 11/37, P = 0.035). CONCLUSION: These findings suggest that: 1) administration of dextrose is associated with nausea, increased opioid requirement and late thirst after elective gynecological laparoscopy; 2) iv fluids did not decrease PONV. PMID- 12734151 TI - Prior ibuprofen exposure does not augment opioid drug potency or modify opioid requirements for pain inhibition in total hip surgery. AB - PURPOSE: In previous animal studies, a prior exposure to non-steroidal anti inflammatory drugs (NSAID) augmented opioid drug potency. This study was designed to answer the question whether a similar effect can be attained in man. The objective was to use NSAID for preoperative pain reduction and at the same time use the NSAID exposure to reduce opioid requirements for pain inhibition in major orthopedic surgery. METHODS: In this double-blind, randomized study, 50 patients scheduled for total hip surgery were included. Patients of Group I received a placebo drug three times a day two weeks before surgery, and those allocated to Group II received ibuprofen (600 mg) three times a day. For surgical anesthesia, all patients received intrathecal bupivacaine 20 mg plus 0.1 mg morphine in a total volume of 4 mL. RESULTS: The preoperative or postoperative visual analogue scale pain scores or the amount of iv morphine showed no differences between the two groups in the first 24 hr after surgery. The median total blood loss in the ibuprofen group was 1161 mL vs 796 mL in the placebo group (P < 0.01). CONCLUSION: Pretreatment with ibuprofen before major hip surgery does not improve the pain scores or reduce morphine requirement but significantly increases blood loss. Considering the presence of relevant adverse effects, pretreatment with a non-selective NSAID is not recommended. PMID- 12734152 TI - Epidural abscess in a patient with dorsal hyperhidrosis. AB - PURPOSE: To report the management of a patient who developed a lumbar epidural abscess when an epidural catheter was placed three years after a thoracic sympathectomy. The possible contribution of hyperhidrosis is discussed. CLINICAL FEATURES: A 62-yr-old male had compensatory hyperhidrosis in his back after thoracic sympathectomy. The patient, who suffered from thromboangeitis obliterans, underwent lumbar (L2-3) epidural catheterization in order to improve arterial circulation and ameliorate resting pain in his left leg. On the third day after catheterization, the patient complained of a dull pain in his back. Emergency magnetic resonance imaging revealed a 12-mm abscess in the epidural space. On the tenth day after catheterization, laminotomy at the 3-4 lumbar vertebrae and local drainage were performed. A 14-mm abscess was removed from the epidural space. The patient was discharged on day 21 after catheterization without any disability. CONCLUSION: Special precautions against infection may be necessary in patients with hyperhidrosis in the area where continuous epidural catheterization is attempted. PMID- 12734154 TI - PDPH is a common complication of neuraxial blockade in parturients: a meta analysis of obstetrical studies. AB - PURPOSE: Postdural puncture headache (PDPH) is an iatrogenic complication of neuraxial blockade. We systematically reviewed the literature on parturients to determine the frequency, onset, and duration of PDPH. METHODS: Citations on PDPH in the obstetrical population were identified by computerized searches, citation review, and hand searches of abstracts and conference proceedings. Citations were included if they contained extractable data on frequency, onset, or duration of PDPH. Using meta-analysis, we calculated pooled estimates of the frequency of accidental dural puncture for epidural needles and pooled estimates of the frequencies of PDPH for epidural and spinal needles. RESULTS: Parturients have approximately a 1.5% [95% confidence interval (CI) 1.5% to 1.5%) risk of accidental dural puncture with epidural insertion. Of these, approximately half (52.1%; 95% CI, 51.4% to 52.8%) will result in PDPH. The risk of PDPH from spinal needles diminishes with small diameter, atraumatic needles, but is still appreciable (Whitacre 27-gauge needle 1.7%; 95% CI, 1.6% to 1.8%). PDPH occurs as early as one day and as late as seven days after dural puncture and lasts 12 hr to seven days. CONCLUSION: PDPH is a common complication for parturients undergoing neuraxial blockade. PMID- 12734153 TI - Systemic, but not pulmonary, hemodynamics are depressed during combined high thoraco-cervical epidural and general anesthesia in dogs. AB - PURPOSE: An epidural block is frequently combined with general anesthesia. Both systemic and pulmonary hemodynamics may be affected by high epidural anesthesia and the combined general anesthetic. These effects were investigated in a canine model. METHODS: Systemic and pulmonary hemodynamics during a combined high thoraco-cervical epidural and general anesthesia were studied in dogs; the animals were anesthetized with propofol, 10 mg.kg(-1).hr(-1), or 2% sevoflurane, and then 1% mepivacaine, 5 mL, was injected epidurally between T1 and T2. Cardiac output (CO), pulmonary capillary wedge pressure (PCWP), pulmonary arterial pressure (PAP), mean arterial pressure (MAP), central venous pressure (CVP), electrocardiogram, and arterial and mixed venous gases were monitored for over 90 min after epidural mepivacaine. The interval between sevoflurane and propofol studies was two hours. RESULTS: Baseline measurement of MAP with sevoflurane anesthesia was significantly lower (P < 0.05-0.01) at every time point than with propofol anesthesia. After epidural mepivacaine (C1)-T7/8 blockade), MAP (P < 0.05-0.01), CO (P < 0.05-0.01), and heart rate (P < 0.05-0.01) decreased significantly during both propofol and sevoflurane anesthesia. In the sevoflurane group, stroke volume decreased significantly (P < 0.05-0.01) but recovered; however, MAP (P < 0.01) and CO (P < 0.05) did not recover 90 min after the injection. Mean CVP and systemic vascular resistance were not altered. There were no changes in mean PAP, mean PCWP, and pulmonary vascular resistance. CONCLUSION: A combined high thoracic/general anesthesia depressed systemic hemodynamics, whereas the pulmonary circulation was not affected. The extent of the depression varied with the general anesthetics used, sevoflurane and propofol. PMID- 12734155 TI - Premedication with nasal s-ketamine and midazolam provides good conditions for induction of anesthesia in preschool children. AB - PURPOSE: To evaluate the efficacy and safety of intranasally administered s ketamine and midazolam for premedication in pediatric patients. METHODS: Ninety children were randomly allocated to receive intranasally administered s-ketamine 1 mg.kg(-1) and midazolam 0.2 mg.kg(-1) (Group K1, n = 30), s-ketamine 2 mg.kg( 1) and midazolam 0.2 mg.kg(-1) (Group K2, n = 30), or midazolam 0.2 mg.kg(-1) (Group M, n = 30) as premedicants, using a double-blind study design. Sedation and anxiolysis were evaluated using a sedation and cooperation scale and recorded at several time points. RESULTS: Acceptable conditions (K1: 23; K2: 26, M: 19) for parental separation were not different between groups. Induction conditions were acceptable in 26 patients in K2 (P < 0.05 vs M) (K1: 23; M: 19). Compared to baseline values individual conditions significantly improved in groups K1 and K2 from 2.5 min after premedication until induction of anesthesia (P < 0.003), in group M conditions improved only five minutes after premedication (P < 0.05). Adverse effects observed in this series were within an acceptable range and similar for the three groups. CONCLUSION: Intranasal administration of s-ketamine and midazolam is an appropriate premedication in preschool children. PMID- 12734156 TI - A background infusion of morphine does not enhance postoperative analgesia after cardiac surgery. AB - PURPOSE: To compare the effects of patient-controlled analgesia (PCA), with or without a background infusion of morphine on postoperative pain relief and stress response after cardiac anesthesia. METHODS: With University Ethics approval, 35 consenting adults undergoing elective open-heart surgery were randomly assigned preoperatively in a double-blind fashion to receive either morphine PCA alone (Group I, n = 15) or morphine PCA plus a continuous basal infusion (Group II, n = 14) for 44 hr postoperatively. Pain scores with visual analogue scale (VAS) at rest, deep inspiration and with cough, sedation scores, stress hormone levels [cortisol, adrenocorticotropin (ACTH) and growth hormone (GH)] and morphine consumption were assessed, and serum morphine levels were measured at four, 20, 28 and 44 hr after surgery. Adverse effects including nausea, vomiting, constipation, urinary retention and pruritus were noted. Total blood, fluid requirements, drainage and urinary output were recorded. RESULTS: Postoperative morphine consumption at 44 hr was less in Group I (29.43 +/- 12.57 mg) than in Group II (50.14 +/- 16.44 mg), P = 0.0006. There was no significant difference between groups in VAS scores, GH levels, blood levels of morphine and adverse effects. While VAS scores, ACTH and GH levels decreased significantly in both groups, plasma cortisol levels increased significantly in Group I only at four hours. In Group II, ACTH and cortisol were higher at four and 44 hr respectively. CONCLUSION: PCA with morphine effectively controlled postoperative pain after cardiac surgery. The addition of a background infusion of morphine did not enhance analgesia and increased morphine consumption. PMID- 12734157 TI - Images in anesthesia: Detection of a defect pulmonary artery catheter balloon by transesophageal echocardiography. PMID- 12734158 TI - Remote preconditioning lessens the deterioration of pulmonary function after repeated coronary artery occlusion and reperfusion in sheep. AB - PURPOSE: We investigated whether remote organ preconditioning (RPC) can preserve pulmonary function following repeated myocardial ischemia/reperfusion in a model mimicking multi-vessel off-pump coronary artery bypass (OPCAB) revascularization. METHODS: Nine sheep (Group-RPC) underwent RPC by three episodes of five-minute occlusion and five-minute reperfusion of the iliac artery. Five sheep (Group-C) were time-matched controls. Afterwards, ten-minute occlusion and reperfusion of the left anterior descending, the first diagonal and the left circumflex coronary arteries were performed consecutively. Hemodynamic and respiratory parameters and arterial blood gases were measured until 120 min after the final coronary reperfusion. Anesthesia was maintained with halothane in oxygen and nitrous oxide. Animals were ventilated with a tidal volume of 15-20 mL.kg(-1) in a non rebreathing system, and a respiratory rate 14-16 min, with 5-cm H(2)O positive end expiratory pressure after thoracotomy. RESULTS: Repeated coronary occlusion and reperfusion was associated in this experimental model with an increase in pulmonary vascular resistance (PVR) and pulmonary arterial pressure (PAP) and a decrease in PaO(2) and PaO(2)/FIO(2) in Group-C. After 120 min reperfusion, PaO(2) and PaO(2)/FIO(2) in Group-RPC were higher (192 +/- 69 mmHg and 241 +/- 78 vs 115 +/- 54 mmHg and 129 +/- 64, P < 0.05), while PVR and PAP were lower than in Group-C. At 120 min of reperfusion, PaO(2) and PaO(2)/FIO(2) were inversely correlated with PVR (P < 0.01). CONCLUSIONS: RPC by transient occlusion of the iliac artery improves lung gas exchange after repeated coronary artery occlusion and reperfusion mimicking OPCAB surgery, and preserves low PVR in sheep. PMID- 12734159 TI - Landiolol, esmolol and propranolol protect from ischemia/reperfusion injury in isolated guinea pig hearts. AB - PURPOSE: Beta blockers are thought to exert beneficial effects on the ischemic heart. The authors examined the effects of landiolol (ONO 1101), a highly selective beta1 antagonist, propranolol, a nonspecific beta blocker, and esmolol, a selective beta1 antagonist, on postischemic contractile recovery. Drugs were given prophylactically. METHODS: Ischemia-reperfusion in isolated guinea pig hearts was induced by stopping the perfusion for 45 min and reperfusing for 60 min. Hearts (n = 7 in each group) were treated with or without propranolol (1 or 10 microM), esmolol (5 or 50 microM), or landiolol (20, 100 or 500 microM) ten minutes before inducing ischemia. RESULTS: At the end of reperfusion, left ventricular pressure (LVP) recovered to 64 +/- 3% of the baseline value in the control group. With 1 and 10 microM propranolol, LVP recovered to 90 +/- 5% and 100 +/- 6% of the baseline value at 60 min after reperfusion, respectively. Fifty microM but not 5 microM of esmolol resulted in restoration of LVP to 97 +/- 17% of the pre-ischemic value at 60 min after reperfusion. In hearts pretreated with 100 and 500 microM landiolol, LVP was restored to 109 +/- 5% and 104 +/- 5% of the baseline value, respectively. Landiolol 100 microM did not depress LVP in the pre-ischemic period. CONCLUSIONS: The present study shows that landiolol, an ultra-short-acting cardioselective beta1 blocker, has cardioprotective effects on ischemia-reperfusion injury in isolated guinea pig hearts. All three beta blockers were equally protective but the intermediate dosage of landiolol preserved LVP during the pre-ischemic period. PMID- 12734160 TI - The PAxpress is an effective ventilatory device but has an 18% failure rate for flexible lightwand-guided tracheal intubation in anesthetized paralyzed patients. AB - PURPOSE: The PAxpress is a new, single-use, extraglottic airway device. We evaluate: 1) insertion success rates; 2) airway sealing pressure, ventilatory capability and calculated mucosal pressures (in vitro minus in vivo intracuff pressure) at 30-60 mL cuff inflation volume; 3) the feasibility of lightwand guided tracheal intubation; and 4) the incidence of mucosal trauma. METHODS: Ninety anesthetized, paralyzed adults were studied. Airway management was by senior anesthesiologists with no prior experience with the PAX, but considerable experience with extraglottic airway devices and the flexible-lightwand. RESULTS: Insertion was successful at the first attempt in 95.5% (86/90) and at the second attempt in 4.5% (4/90). Mean +/- SD airway sealing pressure at 30, 40, 50 and 60 mL cuff inflation volume was 27 +/- 8, 29 +/- 9, 32 +/- 9 and 35 +/- 7 cm H(2)O respectively; expired tidal volume at airway sealing pressure was 16 +/- 6, 18 +/ 6, 19 +/- 5 and 19 +/- 6 mL.kg(-1); and calculated mucosal pressure was 38 +/- 14, 55 +/- 20, 56 +/- 19 and 57 +/- 20 cm H(2)O. Airway sealing pressure, expired tidal volume at airway sealing pressure and calculated mucosal pressures increased with cuff inflation volume (all: P < or = 0.0002). Esophageal leak was detected in 9% (8/90), but only at peak pressures > or = 35 cm H(2)O and cuff inflation volumes > or = 40 mL. Lightwand-guided intubation was successful in 82% (74/90) of patients. Mild, moderate and severe blood staining was detected in 40% (36/90), 15% (13/90) and 1% (1/90) respectively. Blood staining was more frequent after adjusting maneuvers (22/54 vs 32/36, P = 0.002). CONCLUSION: The PAX has a high insertion success rate and is an effective ventilatory device with a low risk of gastric insufflation, but has a moderately high failure rate for lightwand-guided intubation and is associated with a relatively high incidence of mucosal trauma. Mucosal pressures may exceed pharyngeal perfusion pressure. PMID- 12734161 TI - Straight blades improve visualization of the larynx while curved blades increase ease of intubation: a comparison of the Macintosh, Miller, McCoy, Belscope and Lee-Fiberview blades. AB - PURPOSE: To compare the Macintosh (M), McCoy (MC), Miller (MIL), Belscope (BP) and Lee-Fiberview (LF) laryngoscopes with respect to the grade of laryngeal visualization and the difficulty of intubation. METHODS: We included 500 patients scheduled to undergo elective surgery and who required tracheal intubation. Patients were randomly assigned to five groups of 100 patients each. Anesthesia was induced intravenously using 1-3 mg.kg(-1) of propofol, fentanyl 1.5 microg.kg(-1) and atracurium 0.5 mg.kg(-1) or suxamethonium 1 mg.kg(-1). The laryngeal view was classified according to Cormack and Lehane. The degree of difficulty with intubation was rated as: Grade 1, intubation easy; Grade 2, intubation requiring an increased anterior lifting force and assistance to pull the right corner of the mouth upwards to increase space; Grade 3, intubation requiring multiple attempts and a curved stylet; Grade 4, failure to intubate with the assigned laryngoscope. Data were examined using analysis of variance, chi(2) or Fisher test, Student's t test and odds ratio. P < 0.05 was considered statistically significant. RESULTS: Laryngoscopic views obtained with the BP and MIL laryngoscopes were similar, and better than with the other types of laryngoscopes (P < 0.001). The levering tip of the MC blade (P = 0.02) and the fibreoptic device of the LF (P < 0.001) significantly improved the laryngoscopic view. Regarding the degree of difficulty with intubation, the best results were obtained with the MC and M blades (P < 0.001). CONCLUSION: Laryngoscopy was better with straight blades but curved blades provided better intubating conditions. PMID- 12734162 TI - Video-enhanced visualization of the larynx and intubation with the Bullard laryngoscope--equipment report. AB - PURPOSE: A simple technical solution is presented to provide video transmission from the tip of a Bullard laryngoscope to a bedside video display, while the operator is still able to look through the viewing ocular of the Bullard laryngoscope during tracheal intubation. EQUIPMENT: This is achieved by insertion of an ultrathin fibreoptic video-endoscopic system into the working channel of the Bullard laryngoscope. Thereby the view from the distal blade tip is transmitted to a bedside monitor, without interfering with the use of the Bullards laryngoscope's original eyepiece. The presented technical solution allows video transmission without considerable additional weight normally associated with attaching video endoscopy cameras, light and camera cables to endoscopic devices. Thus, the Bullard laryngoscope remains lightweight and easy to maneuver. A screw-threaded adapter with a side-port is proposed to prevent displacement of the fibreoptic cable while still allowing application of oxygen. CONCLUSION: Experience and skills with tracheal intubation using the presented video-enhanced Bullard laryngoscope can be achieved in the originally intended way, while the supervisor or attending viewers can follow the tracheal intubation procedure on the video display. PMID- 12734163 TI - Blind nasotracheal intubation is facilitated by neutral head position and endotracheal tube cuff inflation in spontaneously breathing patients. AB - PURPOSE: To evaluate, using an endotracheal tube mounted on a flexible lighted stylet, how the patient's head-neck position and inflation of the endotracheal cuff affect correct alignment of the tube tip with the glottis. METHODS: Eighty two patients were enrolled. The course of the endotracheal tube in the pharynx was examined by observing the anterior neck for transillumination in each patient under four different intubating conditions. These were: patient's head on pad (8 cm) with the cuff deflated (HP-deflation group); patient's head on pad with the cuff inflated (HP-inflation group); patient's head on bed with the cuff deflated (HB-deflation group); and patient's head on bed with the cuff inflated (HB inflation group). RESULTS: Both a head on bed (neutral) position or endotracheal tube cuff inflation (15 mL of air) significantly increased the frequency of correct alignment of the tip of the endotracheal tube with the glottis. Blind nasotracheal intubation (BNTI) was successful in 69 patients (84%). Lightwand assisted nasotracheal intubation was required in 11 of the remaining 13 patients (13%) and fibreoptic bronchoscopy-assisted intubation was performed in the last two patients. CONCLUSION: A neutral position of the head combined with endotracheal tube cuff inflation is recommended for BNTI. PMID- 12734164 TI - Pain, nausea, vomiting and ocular complications delay discharge following ambulatory microdiscectomy. AB - OBJECTIVE: Nowadays, microsurgical discectomy is being performed as an outpatient procedure. A retrospective chart review was done to document factors that delayed discharge or led to unanticipated admission. METHODS: After Institutional Review Board approval, the hospital medical records of 106 patients who underwent microsurgical discectomy on an ambulatory basis were reviewed. All patients were operated upon by a single surgeon at the Toronto Western Hospital. Perioperative data were collected on specifically designed data sheets. All anesthetic and surgical factors that affected discharge were noted. RESULTS: Of the 106 patients reviewed, only six required unanticipated admission. Two patients were admitted due to nausea and vomiting, one due to severe pain, one due to urinary retention and two were surgical causes (dural tear). Eight patients had delayed discharge. Anesthesia causes were severe nausea, severe pain, low oxygen saturation, sore throat and dry eyes. Two patients had surgical causes. The incidence of postoperative nausea was 61% and postoperative vomiting was 9.4%. Eighty patients (75.4%) complained of pain in the postanesthesia care unit. Of these, 33.9% had visual analogue pain scale scores more than 6. CONCLUSION: Ambulatory lumbar microdiscectomy can be carried out as an ambulatory procedure with an acceptably low unanticipated admission rate (5.7%). The percentage of patients with severe nausea (16%) and pain (33.9%) is high. Adequate perioperative pain management and effective control of nausea and vomiting may further improve the patients' experience after anesthesia for ambulatory microdiscectomy. PMID- 12734165 TI - Postoperative sudden sensorineural hearing loss after posterior lumbar decompression: a case report. AB - PURPOSE: We report a case in which a patient sustained a dural tear during spinal surgery under general anesthesia complicated by a severe and persistent unilateral sensorineural hearing loss. CLINICAL FINDINGS: A 51-yr-old man with no previous otological history underwent a posterior lumbar decompression surgery in the prone position under general anesthesia. A small dural tear was discovered intra-operatively and was repaired by sutures. Surgery lasted for eight hours with over 3 L of blood loss. Recovery from anesthesia was otherwise uneventful. A unilateral right-sided sensorineural hearing loss was discovered shortly after completion of surgery. This was associated with mild tinnitus but no vertigo. There was no aural fullness, pain, headache or postural element to his symptoms. Despite extensive investigations, treatment and follow-up by an otorhinolaryngologist, his symptoms failed to improve 18 months following surgery. DISCUSSION: The possible etiologies, preventive strategies, prognosis and management plan of this rare complication are discussed. PMID- 12734166 TI - Anesthesia: with or without curare? PMID- 12734167 TI - Use of the laryngeal mask airway and a modified sequential intubation technique for the management of an unanticipated difficult airway in a remote location. PMID- 12734168 TI - What do we do with a disconnected epidural catheter? PMID- 12734171 TI - The laryngeal tube and pharyngeal mucosal pressure. PMID- 12734169 TI - Sevoflurane induction of anesthesia in elderly patients. PMID- 12734172 TI - An unusual cause of difficulty confirming correct placement of an endotracheal tube. PMID- 12734174 TI - Propofol decreases cerebral blood flow velocity in anesthetized children. PMID- 12734175 TI - High-degree atrioventricular block after the administration of atropine for sinus arrest during anesthesia. PMID- 12734177 TI - Bioenergetic analysis of peroxisome proliferator-activated receptor gamma coactivators 1alpha and 1beta (PGC-1alpha and PGC-1beta) in muscle cells. AB - Peroxisome proliferator-activated receptor gamma coactivator (PGC)-1alpha is a coactivator of nuclear receptors and other transcription factors that regulates several components of energy metabolism, particularly certain aspects of adaptive thermogenesis in brown fat and skeletal muscle, hepatic gluconeogenesis, and fiber type switching in skeletal muscle. PGC-1alpha has been shown to induce mitochondrial biogenesis when expressed in muscle cells, and preliminary analysis has suggested that this molecule may specifically increase the fraction of uncoupled versus coupled respiration. In this paper, we have performed detailed bioenergetic analyses of the function of PGC-1alpha and its homolog PGC-1beta in muscle cells by monitoring simultaneously oxygen consumption and membrane potential. Cells expressing PGC-1alpha or PGC-1beta display higher proton leak rates at any given membrane potential than control cells. However, cells expressing PGC-1alpha have a higher proportion of their mitochondrial respiration linked to proton leak than cells expressing PGC-1beta. Although these two proteins cause a similar increase in the expression of many mitochondrial genes, PGC-1beta preferentially induces certain genes involved in the removal of reactive oxygen species, recently recognized as activators of uncoupling proteins. Together, these data indicate that PGC-1alpha and PGC-1beta profoundly alter mitochondrial metabolism and suggest that these proteins are likely to play different physiological functions. PMID- 12734178 TI - Type III protein translocase: HrcN is a peripheral ATPase that is activated by oligomerization. AB - Type III protein secretion (TTS) is catalyzed by translocases that span both membranes of Gram-negative bacteria. A hydrophilic TTS component homologous to F1/V1-ATPases is ubiquitous and essential for secretion. We show that hrcN encodes the putative TTS ATPase of Pseudomonas syringae pathovar phaseolicola and that HrcN is a peripheral protein that assembles in clusters at the membrane. A decahistidinyl HrcN derivative was overexpressed in Escherichia coli and purified to homogeneity in a folded state. Hydrodynamic analysis, cross-linking, and electron microscopy revealed four distinct HrcN forms: I, 48 kDa (monomer); II, approximately 300 kDa (putative hexamer); III, 575 kDa (dodecamer); and IV, approximately 3.5 MDa. Form III is the predominant form of HrcN at the membrane, and its ATPase activity is dramatically stimulated (>700-fold) over the basal activity of Form I. We propose that TTS ATPases catalyze protein translocation as activated homo-oligomers at the plasma membrane. PMID- 12734179 TI - Activation of the leptin receptor by a ligand-induced conformational change of constitutive receptor dimers. AB - Binding of leptin to the leptin receptor is crucial for body weight and bone mass regulation in mammals. Leptin receptors were shown to exist as dimers, but the role of dimerization in receptor activation remains unknown. Using a quantitative Bioluminescence Resonance Energy Transfer approach, we show here in living cells that approximately 60% of the leptin receptor exists as constitutive dimers at physiological expression levels in the absence of leptin. No further increase in leptin receptor dimerization was detected in the presence of leptin. Importantly, in cells expressing the short leptin receptor isoform, leptin promoted a robust enhancement of energy transfer signals that reflect specific conformational changes of pre-existing leptin receptor dimers and that may be used as read-out in screening assays for leptin receptor ligands. Both leptin receptor dimerization and the leptin-induced energy transfer were Janus kinase 2 independent. Taken together, our data support a receptor activation model based on ligand-induced conformational changes rather than ligand-induced dimerization. PMID- 12734181 TI - Small conductance Ca2+-activated K+ channels and calmodulin: cell surface expression and gating. AB - Small conductance Ca2+-activated K+ channels (SK channels) are heteromeric complexes of pore-forming alpha subunits and constitutively bound calmodulin (CaM). The binding of CaM is mediated in part by the electrostatic interaction between residues Arg-464 and Lys-467 of SK2 and Glu-84 and Glu-87 of CaM. Heterologous expression of the double charge reversal in SK2, SK2 R464E/K467E (SK2:64/67), did not yield detectable surface expression or channel activity in whole cell or inside-out patch recordings. Coexpression of SK2:64/67 with wild type CaM or CaM1,2,3,4, a mutant lacking the ability to bind Ca2+, rescued surface expression. In patches from cells coexpressing SK2:64/67 and wild type CaM, currents were recorded immediately following excision into Ca2+-containing solution but disappeared within minutes after excision or immediately upon exposure to Ca2+-free solution and were not reactivated upon reapplication of Ca2+-containing solution. Channel activity was restored by application of purified recombinant Ca2+-CaM or exposure to Ca2+-free CaM followed by application of Ca2+-containing solution. Coexpression of the double charge reversal E84R/E87K in CaM (CaM:84/87) with SK2:64/67 reconstituted stable Ca2+ dependent channel activity that was not lost with exposure to Ca2+-free solution. Therefore, Ca2+-independent interactions with CaM are required for surface expression of SK channels, whereas the constitutive association between the two channel subunits is not an essential requirement for gating. PMID- 12734180 TI - Inhibitory effects of insulin-like growth factor-1 and osteogenic protein-1 on fibronectin fragment- and interleukin-1beta-stimulated matrix metalloproteinase 13 expression in human chondrocytes. AB - Matrix metalloproteinase-13 (collagenase-3), a member of the family of matrix metalloproteinases (MMPs), plays a major pathological role in the cartilage destruction of arthritis. A dramatic up-regulation of MMP-13 by inflammatory cytokines such as interleukin (IL)-1beta or by fibronectin fragments has been observed in chondrocytes. In this study, we investigated the inhibitory effects of insulin-like growth factor-1 (IGF-1) and osteogenic protein-1 (OP-1) on the expression of MMP-13, which was induced by fibronectin fragment or IL-1beta in human immortalized or human primary chondrocytes. IGF-1 and OP-1 each significantly reduced the basal level as well as fibronectin fragment- or IL 1beta-stimulated transcription of the MMP-13 gene in a dose-dependent fashion with the corresponding decreases in the protein level of MMP-13. The most prominent suppressive effect was observed by the combination of IGF-1 and OP-1, which decreased the basal promoter activity by 60% and almost completely abrogated the fibronectin fragment-stimulated MMP-13 promoter activity. OP-1 was found to enhance mRNA levels of IGF-1 and the IGF-1 receptor, the latter of which appeared to be responsible for the combined effect of IGF-1 and OP-1. The suppressive effect of IGF-1 and OP-1 on MMP-13 expression was due in part to down regulation of the expression of pro-inflammatory cytokines and the activity of their intermediate molecules, including NF-kappaB and AP-1 factors. We propose that IGF-1 and OP-1 could be key physiological regulators of MMP-13 gene expression and that the combination of IGF-1 and OP-1 may be useful in controlling the excess catabolic activity in arthritis. PMID- 12734182 TI - Use of RNA interference-mediated gene silencing and adenoviral overexpression to elucidate the roles of AKT/protein kinase B isoforms in insulin actions. AB - Insulin plays a central role in the regulation of glucose homeostasis in part by stimulating glucose uptake and glycogen synthesis. The serine/threonine protein kinase Akt has been proposed to mediate insulin signaling in several processes. However, it is unclear whether Akt is involved in insulin-stimulated glucose uptake and which isoforms of Akt are responsible for each insulin action. We confirmed that expression of a constitutively active Akt, using an adenoviral expression vector, promoted translocation of glucose transporter 4 (GLUT4) to plasma membrane, 2-deoxyglucose (2-DG) uptake, and glycogen synthesis in both Chinese hamster ovary cells and 3T3-L1 adipocytes. Inhibition of Akt either by adenoviral expression of a dominant negative Akt or by the introduction of synthetic 21-mer short interference RNA against Akt markedly reduced insulin stimulated GLUT4 translocation, 2-DG uptake, and glycogen synthesis. Experiments with isoform-specific short interference RNA revealed that Akt2, and Akt1 to a lesser extent, has an essential role in insulin-stimulated GLUT4 translocation and 2-DG uptake in both cell lines, whereas Akt1 and Akt2 contribute equally to insulin-stimulated glycogen synthesis. These data suggest a prerequisite role of Akt in insulin-stimulated glucose uptake and distinct functions among Akt isoforms. PMID- 12734184 TI - Origin of the different pH activity profile in two homologous ketosteroid isomerases. AB - Two homologous Delta5-3-ketosteroid isomerases from Comamonas testosteroni (TI WT) and Pseudomonas putida biotype B (PI-WT) exhibit different pH activity profiles. TI-WT loses activity below pH 5.0 due to the protonation of the conserved catalytic base, Asp-38, while PI-WT does not. Based on the structural analysis of PI-WT, the critical catalytic base, Asp-38, was found to form a hydrogen bond with the indole ring NH of Trp-116, which is homologously replaced with Phe-116 in TI-WT. To investigate the role of Trp-116, we prepared the F116W mutant of TI-WT (TI-F116W) and the W116F mutant of PI-WT (PI-W116F) and compared kinetic parameters of those mutants at different pH levels. PI-W116F exhibited significantly decreased catalytic activity at acidic pH like TI-WT, whereas TI F116W maintained catalytic activity at acidic pH like PI-WT and increased the kcat/Km value by 2.5- to 4.7-fold compared with TI-WT at pH 3.8. The crystal structure of TI-F116W clearly showed that the indole ring NH of Trp-116 could form a hydrogen bond with the carboxyl oxygen of Asp-38 like that of PI-WT. The present results demonstrate that the activities of both PI-WT and TI-F116W at low pH were maintained by a tryptophan, which was able not only to lower the pKa value of the catalytic base but also to increase the substrate affinity. This is one example of the strategy nature can adopt to evolve the diversity of the catalytic function in the enzymes. Our results provide insight into deciphering the molecular evolution of the enzyme and creating novel enzymes by protein engineering. PMID- 12734183 TI - Reconstitution of recombinant uncoupling proteins: UCP1, -2, and -3 have similar affinities for ATP and are unaffected by coenzyme Q10. AB - The successful development of recombinant expression and reconstitution protocols has enabled a detailed study of the transport properties and regulation of the uncoupling proteins (UCP). We optimized conditions of isolation and refolding of bacterially expressed uncoupling proteins and reexamined the transport properties and regulation of bacterially expressed UCP1, -2, and -3 reconstituted in liposomes. We show for the first time that ATP inhibits UCP1, -2, and -3 with similar affinities. The Ki values for ATP inhibition were 50 microm (UCP1), 70 microm (UCP2), and 120 microm (UCP3) at pH 7.2. These affinities for ATP are similar to those obtained with native UCP1 isolated from brown adipose tissue mitochondria (Ki = 65 microm at pH 7.2). The Vmax values for proton transport were also similar among the UCPs, ranging from 8 to 20 micromol.min(-1).mg(-1), depending on experimental conditions. We also examined the effect of coenzyme Q on fatty acid-catalyzed proton flux in liposomes containing recombinant UCP1, -2, and -3. We found that coenzyme Q had no effect on the fatty acid-dependent proton transport catalyzed by any of the UCPs nor did it affect nucleotide regulation of the UCPs. We conclude that coenzyme Q is not a cofactor of UCP-mediated proton transport. PMID- 12734185 TI - p53 Activation in adipocytes of obese mice. AB - The tumor suppressor p53 is a transcription factor that activates or represses its target genes after various genotoxic stresses. We have previously shown that sterol regulatory element-binding protein-1 (SREBP-1), a key transcriptional regulator of triglyceride synthesis, and the lipogenic enzymes under its control are markedly suppressed in adipocytes from genetically obese ob/ob mice. Here we demonstrate that p53 and its target genes are highly induced in adipocytes of ob/ob mice in a fed state, leading to the negative regulation of SREBP-1 and thereby lipogenic genes. In fact, disruption of p53 in ob/ob mice completely suppressed the p53-regulated genes to wild-type levels and partially restored expression of lipogenic enzymes. Consistently, reporter gene analysis showed that p53 overexpression suppressed the promoter activity of the SREBP-1c gene and its downstream genes. Thus, the activation of p53 might constitute a negative feedback loop against excess fat accumulation in adipocytes. In conclusion, we discovered a novel role of p53 in the pathophysiology of obesity. PMID- 12734186 TI - Cell-specific and hormone-regulated expression of gonadotropin-regulated testicular RNA helicase gene (GRTH/Ddx25) resulting from alternative utilization of translation initiation codons in the rat testis. AB - Gonadotropin-regulated testicular RNA helicase (GRTH) is a novel DEAD-box protein with ATPase and RNA helicase activities. GRTH gene transcription is stimulated by human chorionic gonadotropin (hCG) via cyclic AMP-induced androgen formation in testicular Leydig cells. In this study, immunocytochemical and Western analyses identified GRTH as a developmentally regulated protein in Leydig cells and in germ cells (pachytene spermatocytes and round spermatids) of the rat testis. Three ATGs with the potential for generation of multiple protein species were identified. Germ cells primarily utilized the 1st ATG codon (+1) and contained major proteins of 61/56 kDa, whereas Leydig cells utilized preferentially the 2nd ATG codon (+ 343) with expression of 48/43 kDa species. A 3rd ATG was weakly utilized and yielded a 33-kDa protein only in germ cells. The increase in GRTH 43 kDa protein in Leydig cells caused by hCG treatment was prevented by the androgen receptor antagonist, flutamide. In round spermatids, hCG caused a significant decrease of 61 kDa species and an induction 48/43 kDa species, whereas no changes were observed in pachytene spermatocytes. Reversal of this hormone-induced switch of expression by flutamide indicated a role of androgen in utilization of the 2nd ATG. These studies have demonstrated a cell-specific and hormone-dependent alternative usage of ATG codons in the testis. They have also revealed that the androgen-dependent transcription of GRTH expression in Leydig cells is accompanied by a marked increase of 43-kDa species. The findings indicate that expression of GRTH proteins is regulated by gonadotropin/androgen at the translational level. PMID- 12734188 TI - A role for the phosphorylation of hRad9 in checkpoint signaling. AB - The integrity of the human genome is preserved by signal transduction pathways called checkpoints, which delay progression through the cell cycle when DNA damage is present. Three checkpoint proteins, hRad9, hRad1, and hHus1, form a proliferating cell nuclear antigen-like, heterotrimeric complex that has been proposed to function in the initial detection of DNA structural abnormalities. hRad9 is highly modified by phosphorylation, in a constitutive manner and in response to both DNA damage and cell cycle position. Here we present evidence that Thr292 of hRad9 is subject to Cdc2-dependent phosphorylation in mitosis. Furthermore, our data are also consistent with four other hRad9 phosphorylation sites (Ser277, Ser328, Ser336, and Thr355) being regulated in part by Cdc2. We also identify Ser387 as a novel site of hRad9 constitutive phosphorylation and show that phosphorylation at Ser387 is a prerequisite for one form of DNA damage induced hyperphosphorylation of hRad9. Characterization of nonphosphorylatable mutants has revealed that hRad9 phosphorylation plays a critical role in checkpoint signaling. Overexpression of these mutants blocks the interaction between hRad9 and the DNA damage-responsive protein TopBP1 and impairs the cellular response to DNA damage during S phase. PMID- 12734187 TI - Src-CrkII-C3G-dependent activation of Rap1 switches growth hormone-stimulated p44/42 MAP kinase and JNK/SAPK activities. AB - We demonstrate here that growth hormone (GH) stimulates the activation of Rap1 and Rap2 in NIH-3T3 cells. Full activation of Rap1 and Rap2 by GH necessitated the combined activity of both JAK2 and c-Src kinases, although c-Src was predominantly required. GH-stimulated Rap1 and Rap2 activity was also demonstrated to be CrkII-C3G-dependent. GH stimulated the tyrosine phosphorylation of C3G, which again required the combined activity of JAK2 and c Src. C3G tyrosine residue 504 was required for GH-stimulated Rap activation. Activated Rap1 inhibited GH-stimulated activation of RalA and subsequent GH stimulated p44/42 MAP kinase activity and Elk-1-mediated transcription. In addition, we demonstrated that C3G-Rap1 mediated CrkII enhancement of GH stimulated JNK/SAPK activity. We have therefore identified a linear JAK2 independent pathway switching GH-stimulated p44/42 MAP kinase and JNK/SAPK activities. PMID- 12734190 TI - Secondary structures common to chloroplast mRNA 3'-untranslated regions direct cleavage by CSP41, an endoribonuclease belonging to the short chain dehydrogenase/reductase superfamily. AB - CSP41 (chloroplast stem-loop-binding protein of 41 kDa), a chloroplast endonuclease belonging to the SDR superfamily, preferentially cleaves stem-loop containing RNAs in vitro. This potentially directs it to the 3'-ends of mature chloroplast mRNAs, which generally possess such structures. To understand the basis for this discrimination, the RNA elements directing CSP41 cleavage of petD RNA in vitro were dissected. Substrates containing fully base-paired stem-loops were optimal substrates, whereas deletion of part of the stem-loop decreased activity by 100-fold, and deletion of the distal arm of the stem-loop abolished cleavage, even in substrates containing the primary CSP41 cleavage site. Competition assays showed that the decrease in activity resulted from decreased affinity for the RNA by CSP41. Mutations of the residues at the scissile bond and mutations and deletions at the terminal loop of the stem had a moderate effect on activity but no effect on cleavage site specificity, suggesting that CSP41 has no sequence specificity. Titration of ethidium bromide into the assay decreased activity to a basal level of approximately 18%, and introduction of a single base bulge into either arm of the stem-loop decreased cleavage at the primary cleavage site by up to 70%. This suggests that changing the structure of the helical stem has a mild effect on activity. Deletion analysis of CSP41 suggests that the specificity domain lies in the first 73 amino acids of the protein, a domain that also contains a putative dehydrogenaselike mononucleotide binding motif. These results are consistent with a broad role for CSP41 in the degradation of stem loop-containing mRNAs. PMID- 12734189 TI - Giardia lamblia RNA polymerase II: amanitin-resistant transcription. AB - Giardia lamblia is an early branching eukaryote, and although distinctly eukaryotic in its cell and molecular biology, transcription and translation in G. lamblia demonstrate important differences from these processes in higher eukaryotes. The cyclic octapeptide amanitin is a relatively selective inhibitor of eukaryotic RNA polymerase II (RNAP II) and is commonly used to study RNAP II transcription. Therefore, we measured the sensitivity of G. lamblia RNAP II transcription to alpha-amanitin and found that unlike most other eukaryotes, RNAP II transcription in Giardia is resistant to 1 mg/ml amanitin. In contrast, 50 microg/ml amanitin inhibits 85% of RNAP III transcription activity using leucyl tRNA as a template. To better understand transcription in G. lamblia, we identified 10 of the 12 known eukaryotic rpb subunits, including all 10 subunits that are required for viability in Saccharomyces cerevisiae. The amanitin motif (amanitin binding site) of Rpb1 from G. lamblia has amino acid substitutions at six highly conserved sites that have been associated with amanitin resistance in other organisms. These observations of amanitin resistance of Giardia RNA polymerase II support previous proposals of the mechanism of amanitin resistance in other organisms and provide a molecular framework for the development of novel drugs with selective activity against G. lamblia. PMID- 12734191 TI - Dwarfism and low insulin-like growth factor-1 due to dopamine depletion in Pts-/- mice rescued by feeding neurotransmitter precursors and H4-biopterin. AB - The tetrahydrobiopterin (BH4) cofactor is essential for the biosynthesis of catecholamines and serotonin and for nitric-oxide synthase (NOS). Alterations in BH4 metabolism are observed in various neurological and psychiatric diseases, and mutations in one of the human metabolic genes causes hyperphenylalaninemia and/or monoamine neurotransmitter deficiency. We report on a knockout mouse for the Pts gene, which codes for a BH4-biosynthetic enzyme. Homozygous Pts-/- mice developed with normal morphology but died after birth. Upon daily oral administration of BH4 and neurotransmitter precursors the Pts-/- mice eventually survived. However, at sexual maturity (6 weeks) the mice had only one-third of the normal body weight and were sexually immature. Biochemical analysis revealed no hyperphenylalaninemia, normal brain NOS activity, and almost normal serotonin levels, but brain dopamine was 3% of normal. Low dopamine leads to impaired food consumption as reflected by the severe growth deficiency and a 7-fold reduced serum insulin-like growth factor-1 (IGF-1). This is the first link shown between 6-pyruvoyltetrahydropterin synthase- or BH4-biosynthetic activity and IGF-1. PMID- 12734192 TI - Dominant-negative cAMP-responsive element-binding protein inhibits proliferating cell nuclear antigen and DNA repair, leading to increased cellular radiosensitivity. AB - Selective inhibition of the epidermal growth factor receptor or mitogen-activated protein kinase (MAPK) results in radiosensitization of cancer cells. One potential mechanism involves cAMP-responsive element-binding protein, which is activated by radiation via the epidermal growth factor receptor/MAPK pathway and which regulates synthesis of proliferating cell nuclear antigen (PCNA), a protein involved in repair of ionizing radiation-induced DNA damage. To test for a role of CREB in cellular radiosensitivity, CHO cells were transfected with plasmids expressing dominant-negative CREB mutants (CR133 or KCREB), and various end points were measured 48 h later. Basal levels of PCNA-CAT reporter construct activity were reduced by 60 and 40% following expression of CR133 and KCREB, respectively; similar decreases were observed in PCNA protein levels. Pulsed field gel electrophoresis measurements showed that CR133 inhibited the repair of radiation-induced DNA double-strand breaks, and this effect was reversed by over expression of PCNA; dominant-negative CREB also significantly inhibited split dose recovery. Clonogenic assays were used to determine surviving fraction; the dose enhancement ratios for dominant-negative CREB-expressing cells compared with control (vector alone) were 1.5 and 1.3 for CR133 and KCREB, respectively. Importantly, co-transfection of mutant CREB and a construct constitutively expressing PCNA protein restored radiosensitivity of CHO cells back to wild-type levels. Moreover, cells expressing either CREB mutant showed no significant cell cycle redistribution. These data demonstrate that genetic disruption of CREB results in radiosensitization, and that this effect can be explained by a mechanism involving decreased PCNA expression and inhibition of DNA repair. PMID- 12734193 TI - Transforming growth factor-beta-Smad signaling pathway negatively regulates nontypeable Haemophilus influenzae-induced MUC5AC mucin transcription via mitogen activated protein kinase (MAPK) phosphatase-1-dependent inhibition of p38 MAPK. AB - In contrast to the extensive studies on the role of transforming growth factor beta (TGF-beta) in regulating cell proliferation, differentiation, and apoptosis over the past decade, relatively little is known about the exact role of TGF-beta signaling in regulating host response in infectious diseases. Most of the recent studies have suggested that TGF-beta inhibits macrophage activation during infections with pathogens such as Trypanosoma cruzi and Leishmania, thereby favoring virulence. In certain situations, however, there is also evidence that TGF-beta has been correlated with enhanced resistance to microbes such as Candida albicans, thus benefiting the host. Despite these distinct observations that mainly focused on macrophages, little is known about how TGF-beta regulates host primary innate defensive responses, such as up-regulation of mucin, in the airway epithelial cells. Moreover, how the TGF-beta-Smad signaling pathway negatively regulates p38 mitogen-activated protein kinase (MAPK), a key pathway mediating host response to bacteria, still remains largely unknown. Here we show that nontypeable Haemophilus influenzae, a major human bacterial pathogen of otitis media and chronic obstructive pulmonary diseases, strongly induces up-regulation of MUC5AC mucin via activation of the Toll-like receptor 2-MyD88-dependent p38 path-way. Activation of TGF-beta-Smad signaling, however, leads to down regulation of p38 by inducing MAPK phophatase-1, thereby acting as a negative regulator for MUC5AC induction. These studies may bring new insights into the novel role of TGF-beta signaling in attenuating host primary innate defensive responses and enhance our understanding of the signaling mechanism underlying the cross-talk between TGF-beta-Smad signaling pathway and the p38 MAPK pathway. PMID- 12734194 TI - The glutathione synthetase of Schizosaccharomyces pombe is synthesized as a homodimer but retains full activity when present as a heterotetramer. AB - Glutathione synthetase was overexpressed as a histidine-tagged protein in Schizosaccharomyces pombe and purified by two-step affinity chromatography. The recovered enzyme occurred in two different forms: a homodimeric protein consisting of two identical 56-kDa subunits and a heterotetrameric protein composed of two 32-kDa and two 24-kDa subfragments. Both forms are encoded by the GSH2 gene. The 56-Da protein corresponds to the complete GSH2 open reading frame, while the subfragments are produced following the cleavage of this larger protein by a metalloprotease. A stable homodimer was obtained by site-directed mutagenesis to remove the protease cleavage site, and this showed normal activity. A structural model of the fission yeast glutathione synthetase was produced, based on the x-ray coordinates of the human enzyme. According to this model the interacting domains of the proteolytic subfragments are strongly entangled. The subfragments were therefore coexpressed as independent proteins. These subfragments assembled correctly to yield functional heterotetramers with equivalent activity to the wild type enzyme. Furthermore, a permuted version of the protein was created. This also showed normal levels of glutathione synthetase activity. These data provide novel insight into the mechanisms of protein folding and the structure and evolution of the glutathione synthetase family. PMID- 12734197 TI - Evidence for distinct roles in catalysis for residues of the serine-serine-lysine catalytic triad of fatty acid amide hydrolase. AB - Fatty acid amide hydrolase (FAAH) is a mammalian amidase signature enzyme that inactivates neuromodulatory fatty acid amides, including the endogenous cannabinoid anandamide and the sleep-inducing substance oleamide. The recent determination of the three-dimensional structures of FAAH and two distantly related bacterial amidase signature enzymes indicates that these enzymes employ an unusual serine-serine-lysine triad for catalysis (Ser-241/Ser-217/Lys-142 in FAAH). Mutagenesis of each of the triad residues in FAAH has been shown to severely reduce amidase activity; however, how these residues contribute, both individually and in cooperation, to catalysis remains unclear. Here, through a combination of site-directed mutagenesis, enzyme kinetics, and chemical labeling experiments, we provide evidence that each FAAH triad residue plays a distinct role in catalysis. In particular, the mutation of Lys-142 to alanine indicates that this residue functions as both a base involved in the activation of the Ser 241 nucleophile and an acid that participates in the protonation of the substrate leaving group. This latter property appears to support the unusual ability of FAAH to hydrolyze amides and esters at equivalent rates. Interestingly, although structural evidence indicates that the impact of Lys-142 on catalysis probably occurs through the bridging Ser-217, the mutation of this latter residue to alanine impaired catalytic activity but left the amide/ester hydrolysis ratios of FAAH intact. Collectively, these findings suggest that FAAH possesses a specialized active site structure dedicated to a mechanism for competitive amide and ester hydrolysis where nucleophile attack and leaving group protonation occur in a coordinated manner dependent on Lys-142. PMID- 12734196 TI - Characterization of cadherin-24, a novel alternatively spliced type II cadherin. AB - Cadherins comprise a superfamily of calcium-dependent cell-cell adhesion molecules. Within the superfamily are six subfamilies including type I and type II cadherins. Both type I and type II cadherins are composed of five extracellular repeat domains with conserved calcium-binding motifs, a single pass transmembrane domain, and a highly conserved cytoplasmic domain that interacts with beta-catenin and p120 catenin. In this study, we describe a novel cadherin, cadherin-24. It is a type II cadherin with a 781-codon open reading frame, which encodes a type II cadherin protein complete with five extracellular repeats containing calcium-binding motifs, a transmembrane domain, and a conserved cytoplasmic domain. Cadherin-24 has the unusual feature of being alternatively spliced in extracellular repeat 4. This alternative exon encodes 38 in-frame amino acids, resulting in an 819-amino-acid protein. Sequence analysis suggests the presence of beta-catenin and p120 catenin-binding sequences, and immunoprecipitation experiments confirm the ability of both forms of the novel cadherin to associate with alpha-catenin, beta-catenin, and p120 catenin. In addition, aggregation assays show that both forms of cadherin-24 mediate strong cell-cell adhesion. PMID- 12734198 TI - Gene modulation by the cyclooxygenase inhibitor, sulindac sulfide, in human colorectal carcinoma cells: possible link to apoptosis. AB - The mechanisms underlying the anti-tumorigenic properties of cyclooxygenase inhibitors are not well understood. One novel hypothesis is alterations in gene expression. To test this hypothesis sulindac sulfide, which is used to treat familial adenomatous polyposis, was selected to detect gene modulation in human colorectal cells at physiological concentrations with microarray analysis. At micromolar concentrations, sulindac sulfide stimulated apoptosis and inhibited the growth of colorectal cancer cells on soft agar. Sulindac sulfide (10 microm) altered the expression of 65 genes in SW-480 colorectal cancer cells, which express cyclooxygenase-1 but little cyclooxygenase-2. A more detailed study of 11 genes revealed that their expression was altered in a time- and dose-dependent manner as measured by real-time RT-PCR. Northern analysis confirmed the expression of 9 of these genes, and Western analysis supported the conclusion that sulindac sulfide altered the expression of these proteins. Cyclooxygenase deficient HCT-116 cells were more responsive to sulindac sulfide-induced gene expression than SW-480 cells. However, this response was diminished in HCT-116 cells overexpressing cyclooxygenase-1 compared with normal HCT-116 cells suggesting the presence of cyclooxygenase attenuates this response. However, prostaglandin E2, the main product of cyclooxygenase, only suppressed the sulindac sulfide-induced expression of two genes, with little known biological function while it modulated the expression of two more. The most likely explanation for this finding is the metabolism of sulindac sulfide to inactive metabolites by the peroxidase activity of cyclooxygenase. In conclusion, this is the first report showing sulindac sulfide, independent of cyclooxygenase, altered the expression of several genes possibly linked to its anti-tumorigenic and pro apoptotic activity. PMID- 12734199 TI - Osteoblast-derived oxysterol is a migration-inducing factor for human breast cancer cells. AB - Bone metastasis is the major reason for death caused by breast cancer. We used human breast cancer (MCF-7) cells that are poorly metastatic but show highly inducible migration to determine bone-derived factors that induce migration of initially non-disseminating breast cancer cells. We have found that a lipid fraction from human osteoblast-like MG63 cell-conditioned medium (MG63CM) contains a migration-inducing factor for MCF-7 cells. In this fraction, we have identified oxysterol (OS) as a lipid mediator for tumor cell migration. In MCF-7 cells, insulin-like growth factor 1 elevates the expression of OS-binding protein related protein 7. Binding of OS to OS-binding protein or OS-binding protein related protein is known to trigger elevation of sphingomyelin, a sphingolipid that organizes lipid microdomains in the cell membrane. In MCF-7 cells, OS increases the intracellular concentration of sphingomyelin and other phospholipids and induces the translocation of the small GTPase p21Ras to GM1- and cholesterol-rich membrane areas. The induction of migration by MG63CM is prevented by incubation of MG63 cells with mevinolin, a statin-type cholesterol biosynthesis inhibitor that depletes the conditioned medium of OS. Osteoblast derived OS may, thus, be a yet unrecognized lipid mediator for bone metastasis of breast cancer and a new target for anti-metastasis chemotherapy with statins. PMID- 12734200 TI - Characterization of the oligosaccharides associated with the human ovarian tumor marker CA125. AB - CA125 is a mucin commonly employed as a diagnostic marker for epithelial ovarian cancer. Induction of humoral responses to CA125 leads to increased survival times in patients with this form of cancer, suggesting a potential role for this mucin in tumor progression. In this study, oligosaccharides linked to CA125 derived from the human ovarian tumor cell line OVCAR-3 were subjected to rigorous biophysical analysis. Sequencing of the O-glycans indicates the presence of both core type 1 and type 2 glycans. An unusual feature is the expression of branched core 1 antennae in the core type 2 glycans. CA125 is also N-glycosylated, expressing primarily high mannose and complex bisecting type N-linked glycans. High mannose type glycans include Man5-Man9GlcNAc2. The predominant N-glycans are the biantennary, triantennary, and tetraantennary bisecting type oligosaccharides. Remarkably, the N-glycosylation profiles of CA125 and the envelope glycoprotein gp120 (derived from H9 lymphoblastoid cells chronically infected with HIV-1) are very similar. The CA125-associated N-glycans have also recently been implicated in crucial recognition events involved in both the innate and adaptive arms of the cell-mediated immune response. CA125 may therefore induce specific immunomodulatory effects by employing its carbohydrate sequences as functional groups, thereby promoting tumor progression. Immunotherapy directed against CA125 may attenuate these immunosuppressive effects, leading to the prolonged survival of patients with this extremely serious form of cancer. PMID- 12734201 TI - The base substitution fidelity of DNA polymerase beta-dependent single nucleotide base excision repair. AB - Damaged DNA bases are removed from mammalian genomes by base excision repair (BER). Single nucleotide BER requires several enzymatic activities, including DNA polymerase and 5',2'-deoxyribose-5-phosphate lyase. Both activities are intrinsic to four human DNA polymerases whose base substitution error rate during gap filling DNA synthesis varies by more than 10,000-fold. This suggests that BER fidelity could vary over a wide range in an enzyme dependent manner. To investigate this possibility, here we describe an assay to measure the fidelity of BER reactions reconstituted with purified enzymes. When human uracil DNA glycosylase, AP endonuclease, DNA polymerase beta, and DNA ligase 1 replace uracil opposite template A or G, base substitution error rates are or =10% of the original CFU for longer than all the other swabs. In the clinical study, the ability of the Cellswab to detect beta hemolytic streptococci from throat samples (n = 995) was compared to that of the TSC Dacron swab. The swabs performed equally, both when their samples were transferred to plates immediately and after storage for 1 day at 4 degrees C or RT. The changes in normal microbiota after storage were also similar. The Cellswab was found to perform at least as well as ordinary swabs. It was better at storing fastidious strains, and at keeping bacteria viable for long storage times; it might well be a useful replacement or complement to ordinary swabs. PMID- 12734222 TI - Multicenter evaluation of use of dried blood and plasma spot specimens in quantitative assays for human immunodeficiency virus RNA: measurement, precision, and RNA stability. AB - Eleven laboratories evaluated the use of dried blood and plasma spots for quantitation of human immunodeficiency virus (HIV) RNA by two commercially available RNA assays, the Roche Amplicor HIV-1 Monitor and the bioMerieux NucliSens HIV-1 QT assays. The recovery of HIV RNA was linear over a dynamic range extending from 4,000 to 500,000 HIV type 1 RNA copies/ml. The Monitor assay appeared to have a broader dynamic range and seemed more sensitive at lower concentrations. However, the NucliSens assay gave more consistent results and could be performed without modification of the kit. HIV RNA was stable in dried whole blood or plasma stored at room temperature or at -70 degrees C for up to 1 year. Dried blood and dried plasma spots can be used as an easy and inexpensive means for the collection and storage of specimens under field conditions for the diagnosis of HIV infection and the monitoring of antiretroviral therapy. PMID- 12734224 TI - New enzyme immunoassay for detection of hepatitis B virus core antigen (HBcAg) and relation between levels of HBcAg and HBV DNA. AB - A new enzyme immunoassay specific for hepatitis B virus (HBV) core antigen (HBcAg) was developed. In order to detect HBcAg, specimens were pretreated with detergents to release HBcAg from the HBV virion and disassemble it to dimers, and simultaneously, the treatment inactivated anti-HBc antibodies. HBcAg detected by the assay peaked with HBV DNA in density gradient fractions of HBV-positive sera. The assay showed a wide detection range from 2 to 100,000 pg/ml. We observed no interference from anti-HBc antibody or blood components, but the assay was inhibited by very high concentrations (>1 microg/ml; corresponding to 80 signal/cutoff) of HBeAg. When the cutoff value was tentatively set at 4 pg/ml, all healthy control (HBsAg and HBV DNA negative, n = 160) and anti-hepatitis C virus-positive (n = 55) sera were identified as negative. HBcAg concentrations correlated very closely with HBV DNA (r = 0.946, n = 145) in 216 samples from 72 hepatitis B patients. In seroconversion panels, HBcAg concentrations changed in parallel with HBV DNA levels. The assay, therefore, offers a simple method for monitoring hepatitis B patients. With a series of sera during lamivudine therapy, HBV DNA levels fell sharply and the HBcAg concentration also decreased, but the change in HBcAg was smaller and more gradual. The supposed mechanism of these changes and their clinical significance are discussed. PMID- 12734225 TI - Candida dubliniensis at a university hospital in Saudi Arabia. AB - Candida dubliniensis is a newly described yeast species that is a close phylogenetic relative of C. albicans. Although it has been reported from different parts of the world, no detailed investigation of this species has been done in Saudi Arabia. The purpose of the present study was to identify C. dubliniensis isolates recovered from clinical specimens at a tertiary-care hospital in Riyadh, Saudi Arabia, and to determine the drug susceptibility profiles of those isolates. Over a period of 8 months, 823 germ tube- and chlamydospore-positive yeasts identified as C. albicans and recovered from different clinical specimens were screened for their ability to grow at 45 degrees C on Sabouraud dextrose agar. Isolates which failed to grow at 45 degrees C were presumptively identified as C. dubliniensis. The species identities were further confirmed by the production of pseudohyphae and chlamydospores on Staib agar and their inability to assimilate D-xylose and alpha-methyl-D-glucoside by using the API 20C AUX system. A total of 27 (3.3%) isolates were identified as C. dubliniensis. They were all recovered from 23 human immunodeficiency virus negative patients. The prevalence of C. dublinensis in bronchoalveolar lavage (33.3%), oral (16.7%), and blood (16.7%) specimens was high. In addition, 33 isolates previously identified as C. albicans and preserved among our stock blood culture isolates were also recruited for the study. Of these, 5 isolates were found to be C. dubliniensis, thus making the total number of isolates identified as this species 32. Antifungal susceptibility testing of the C. dubliniensis isolates showed 100% sensitivity to amphotericin B, 97% sensitivity to each of fluconazole and ketoconazole, and 87.5% sensitivity to itraconazole. However, in contrast to other studies, the majority of the isolates (65.6%) showed high levels of resistance to flucytosine (MIC > 64 microg/ml). Further studies are warranted to investigate the cause of this unusually high rate of resistance to flucytosine of the C. dubliniensis isolates in this region. PMID- 12734226 TI - Quality control for beta-lactam susceptibility testing with a well-defined collection of Enterobacteriaceae and Pseudomonas aeruginosa strains in Spain. AB - Eighteen Enterobacteriaceae and Pseudomonas aeruginosa strains, 16 of them with well-defined beta-lactam resistance mechanisms, were sent to 52 Spanish microbiology laboratories. Interpretative categories for 8 extended-spectrum beta lactams were collected. Participating laboratories used their own routine susceptibility testing procedures (88% automatic systems, 10% disk diffusion, and 2% agar dilution). Control results were established by two independent reference laboratories by applying the NCCLS microdilution method and interpretative criteria. Interpretative discrepancies were observed in 16% of the results (4.4% for cefepime, 3.0% for aztreonam, 2.8% for piperacillin-tazobactam, 1.7% for cefotaxime [CTX] and ceftazidime, 1.1% for ceftriaxone, 0.9% for meropenem, and 0.3% for imipenem). High consistency with reference values (<5% of major plus very major errors) was observed with (i) American Type Culture Collection quality control strains; (ii) strains with low-efficiency mechanisms inactivating extended-spectrum beta-lactams, such as OXA-1-producing Escherichiacoli or SHV-1 hyperproducing Klebsiella pneumoniae; (iii) strains with highly efficient mechanisms, such as SHV-5 porin-deficient K. pneumoniae, CTX-M-10 in Enterobacter cloacae hyperproducing AmpC, and P. aeruginosa with the MexAB OprM efflux phenotype or hyperproducing AmpC. Low consistency (>30% major plus very major errors) was detected in K1-producing Klebsiella oxytoca, CTX-M-9-producing E. coli, and in OprD(-) P. aeruginosa strains. Extended-spectrum beta-lactamase (ESBL)-producing strains accounted for 86% of very major errors. Recognition of the ESBL phenotype was particularly low in Enterobacter cloacae strains (<35%), due to the lack of NCCLS-specific rules in this genus. A K1-producing K. oxytoca was misidentified by 10% of laboratories as an ESBL producer. The use of well defined resistant strains is useful for improving proficiency in susceptibility testing in clinical laboratories. PMID- 12734227 TI - Antimicrobial resistance and genetic diversity of Shigella sonnei isolates from western Ireland, an area of low incidence of infection. AB - Shigella sonnei is a significant cause of gastroenteritis in both developing and industrialized countries. Definition of the diversity and antimicrobial susceptibility of S. sonnei isolates may be helpful in the management of individual cases and outbreaks. Antimicrobial susceptibility testing and pulsed field gel electrophoresis (PFGE) were performed with 67 isolates of S. sonnei predominantly (n = 59) from three counties in the west of Ireland. Phage typing (n = 17), plasmid profiling (n = 28), and integron analysis (n = 24) were performed with subsets of strains. PFGE typing permitted recognition of two major clusters: PFGE type A (n = 53) and PFGE type B (n = 14). PFGE type A was associated with resistance to ampicillin, streptomycin, and sulfonamides (51 of 53 isolates), and those that were phage typed (n = 6) were phage type 3. PFGE type B was associated with resistance to streptomycin, sulfonamides, tetracycline, and trimethoprim (11 of 14 isolates) and phage type 6 (9 of 11 isolates). Fifteen different plasmid profiles were identified among the 28 isolates analyzed. A class 2 integron was present in all 14 PFGE type B isolates. One of these isolates also contained a class 1 integron and showed a unique variant of the PFGE type B pattern. Sequence analysis of the gene cassette structures contained within these integrons identified distinct open reading frames that encoded determinants of resistance to trimethoprim, streptomycin, and streptothricin. Our data demonstrate two predominant PFGE types among S. sonnei isolates circulating in this region. The limited diversity of the S. sonnei isolates in this region means that detection of isolates indistinguishable by PFGE and according to their antibiograms in two or more patients is not persuasive evidence of a common-source food- or waterborne outbreak. Indistinguishable plasmid profiles in addition to indistinguishable PFGE and antibiogram types may be more suggestive of an epidemiologically relevant link between cases. PMID- 12734228 TI - BVBlue test for diagnosis of bacterial vaginosis. AB - Bacterial vaginosis (BV) is a disorder of the vaginal ecosystem characterized by a shift in the vaginal flora from the normally predominant Lactobacillus to one dominated by sialidase enzyme-producing mixed flora. It is the most common cause of abnormal vaginal discharge in adult women. The BVBlue system (Gryphus Diagnostics, L.L.C.) is a chromogenic diagnostic test based on the presence of elevated sialidase enzyme in vaginal fluid samples. BVBlue was compared to the standard method for diagnosing BV (Amsel criteria and Nugent score). Fifty-seven nonmenstruating women of > or =16 years of age who presented for a pelvic examination were recruited. Demographic features were collected via a self administered questionnaire. The Amsel criteria were assessed based on three of four of the following characteristics of vaginal discharge: consistency, odor, pH, and presence of clue cells on Gram stain. BVBlue was compared to the Gram stain and Amsel criteria. The sensitivity, specificity, positive predictive value, and negative predictive value for BVBlue versus the Gram stain and Amsel criteria were 91.7, 97.8, 91.7, and 97.8% and 50.0, 100, 100, and 88.2%, respectively. A significantly greater proportion of patients with a vaginal pH of >4.5, a positive amine test, or with clue cells on vaginal Gram smear were found to have a positive BVBlue test (P < 0.001). Women previously treated for BV were 2.98 times more likely to have another episode of BV. BVBlue is a useful point-of care diagnostic tool to provide a presumptive diagnosis of BV, especially in situations where microscopic capabilities are unavailable. PMID- 12734229 TI - Escherichia coli strains from pregnant women and neonates: intraspecies genetic distribution and prevalence of virulence factors. AB - To determine the extent to which the vagina, endocervix, and amniotic fluid screen the Escherichia coli strains responsible for neonatal infections, we studied the genetic relationships among 105 E. coli strains isolated from all of the ecosystems involved in this infectious process. Twenty-four strains were isolated from the intestinal flora, and 25 strains were isolated from the vaginas of pregnant women. Twenty-seven strains were isolated from the amniotic fluid, blood, and cerebrospinal fluid (CSF) of infected neonates. The intraspecies genetic characteristics of all of the isolates were determined by random amplified polymorphic DNA (RAPD) analysis, PCR ECOR (E. coli reference) grouping, and PCR virulence genotyping. A correlation was found between the intraspecies distributions of the strains in the A, B1, B2, and D ECOR groups and in the two major RAPD groups (I and II). Nevertheless, the distribution of the E. coli strains in the RAPD groups according to their anatomical origins was more significant than their distribution in the ECOR groups. This may be explained by the existence of an E. coli subpopulation, defined by the RAPD I group, within the ECOR B2 group. This RAPD I group presents a major risk for neonates: 75% of the strains isolated from patients with meningitis and 100% of the strains isolated from patients with bacteremia were in this group. The vagina and the amniotic fluid are two barriers that favor colonization by highly infectious strains. Indeed, only 17% of fecal strains belonged to the RAPD I group, whereas 52% of vaginal strains and 67% of amniotic fluid strains belonged to this subpopulation. The ibeA and iucC genes were significantly associated with CSF strains, whereas the hly and sfa/foc genes were more frequent in blood strains. These findings could serve as a basis for developing tools to recognize vaginal strains, which present a high risk for neonates, for use in prophylaxis programs. PMID- 12734230 TI - Comparison of the pathogenesis of two genetically different H3N2 influenza A viruses in pigs. AB - In 1997 and 1998, H3N2 influenza A viruses emerged among pigs in North America. Genetic analyses of the H3N2 isolates demonstrated that they had distinctly different genotypes. The most commonly isolated viruses in the United States have a triple-reassortant genotype, with the hemagglutinin, neuraminidase, and PB1 polymerase genes being of human influenza virus origin, the nucleoprotein, matrix, and nonstructural genes being of classical swine influenza virus origin, and the PA and PB2 polymerase genes being of avian influenza virus origin. In contrast, a wholly human H3N2 virus was isolated from a single baby pig in Ontario, Canada, in 1997, but it did not spread within the swine population. Genetic differences between this wholly human virus and the triple-reassortant viruses may affect their replication efficiencies in pigs. In the present study we compared the pathogenicities and replication kinetics of the wholly human virus and a triple-reassortant virus in 7-week-old pigs that were infected intranasally with 2 x 10(3) to 2 x 10(6) 50% tissue culture infective doses of virus. Our results demonstrate that the wholly human virus replicated to significantly lower titers and that the onset of virus shedding was delayed compared to the replication titers and the time of onset of virus shedding in triple-reassortant viruses. In addition, infection with the triple-reassortant virus was associated with moderate to severe gross pathological and histological pulmonary lesions, while infection with the wholly human virus induced only mild pulmonary changes. PMID- 12734231 TI - Regional variation among vacA alleles of Helicobacter pylori in China. AB - Allelic variation among Helicobacter pylori vacA occurs in the signal and middle region of the gene. The aim of the study was to investigate alleleic variation of vacA among H. pylori strains from three regional areas of China and the relationship between vacA alleles and PUD. DNA extracted from 88 clinical isolates of H. pylori was analyzed by type-specific PCR and reverse hybridization line probe assay (LiPA) to determine the genotype of vacA and presence of cagA. In 87 isolates, all of the vacA alleles could be classified as either type s1c or type s1a, and all could be classified as m1, m2a, or m2b. One strain could not be typed. In all, 41% of patients were infected with multiple vacA genotypes, with the highest level being observed in Shanghai (63%). In strains from Beijing, s1a was dominant; by contrast, s1c was dominant in Guangxi and Shanghai. The prevalence of m2b strains in Shanghai (63%) was significantly higher than that in Beijing (3%) or Guangxi (0%). Thirty of the 87 patients had peptic ulcers. However, there was no association between vacA genotype and PUD. This study demonstrates that there is significant geographic diversity of genotype of vacA within China. The absence of vacA s2 genotypes precluded analysis of an association of vacA s genotypes and clinical disease. PMID- 12734232 TI - Genomic diversity of clinical and environmental Vibrio cholerae strains isolated in Brazil between 1991 and 2001 as revealed by fluorescent amplified fragment length polymorphism analysis. AB - Vibrio cholerae is a ubiquitous and abundant organism in aquatic environments, particularly in coastal areas, estuaries, and rivers. This organism was the cause of a considerable number of deaths in Brazil during the last decade. In this study we applied the genomic fingerprinting technique fluorescent amplified fragment length polymorphism (FAFLP) to analyze 106 V. cholerae O1 and non-O1 and non-O139 strains isolated from clinical specimens and the environment between 1991 and 2001. Numerical analysis of the FAFLP patterns disclosed seven main groups of genomes, all of them originated from a variety of different places in different years, suggesting that V. cholerae is a very diverse species. O1 and non-O1 and non-O139 strains were distinguishable by FAFLP, although clinical and environmental strains clustered together in a few cases. The persistence of some strains of highly related genomes during several years and in completely different geographical regions suggests that these strains are highly successful in adapting to changing environmental conditions. PMID- 12734233 TI - Validation of the automated reading and incubation system with Sensititre plates for antimicrobial susceptibility testing. AB - The present study compared the antimicrobial susceptibility testing (AST) results generated by the Automated Incubation and Reading System (ARIS) with custom Sensititre plates (TREK Diagnostic Systems, Cleveland, Ohio) and MicroScan PC10 GP and NUMIC10 GN plates interpreted with the WalkAway-96 system (Dade Behring, West Sacramento, Calif.) for gram-positive (GP) and gram-negative (GN) organisms as part of an in-house validation. A total of 326 isolates (3,689 antimicrobial agent-organism combinations) were evaluated. Sensititre plates were inoculated according to the instructions of the manufacturer with a suspension adjusted to a 0.5 McFarland standard, while the Prompt Inoculation System was used for the MicroScan plates. ARIS and the WalkAway system were used for automated reading of the Sensititre and MicroScan plates, respectively, at 18 to 24 h. The results were analyzed for essential (+/-1 twofold dilution) and categorical (sensitive, intermediate, or resistant) agreements. Plates that resulted in ARIS interpretations with major (falsely resistant) or very major (falsely susceptible) errors compared to the results obtained with the WalkAway system were read manually to corroborate instrument readings. Isolates for which very major or major errors were obtained and for which the results were not resolved by manual reading were retested in parallel. Isolates for which very major or major errors were obtained and for which the results were not resolved upon repeat testing were tested by the National Committee for Clinical Laboratory Standards M7-A5 frozen reference microdilution method. Essential agreement was 95.8% for 246 GN isolates. The following categorical error rates were obtained for the GN isolates: 1.3% minor errors, 0% major errors, and 0.4% very major errors. For 95 GP isolates, there was 93.5% essential agreement. Categorical error rates for GP isolates were 0.9% minor errors, 0.6% major errors, and 0.4% very major errors. ARIS-Sensititre is a diagnostic system feasible for use for automated AST in a clinical laboratory. PMID- 12734234 TI - Detection of Chlamydia pneumoniae-specific antibodies binding to the VD2 and VD3 regions of the major outer membrane protein. AB - Although Chlamydia pneumoniae is an important human pathogen, the antigens eliciting a specific humoral immune response remain elusive. We scrutinized several recombinant chlamydial surface proteins for species-specific recognition by a panel of human sera previously tested for the presence of anti-C. pneumoniae and anti-C. trachomatis antibodies by microimmunofluorescence and enzyme-linked immunosorbent assay. The 15-kDa cysteine-rich protein (CrpA), porin-b (PorB), 9 kDa outer membrane protein (OMP3), 60-kDa outer membrane protein (OMP2), and four fragments of the major outer membrane protein (MOMP) representing each variable domain (VD) were overexpressed in Escherichia coli, affinity purified, and employed for Western blot analysis. None of the sera tested contained antibodies recognizing PorB and OMP3 of C. pneumoniae. Sera from C. pneumoniae-immune patients cross-reacted with OMP2 of C. trachomatis, and sera from C. trachomatis immune patients cross-reacted with CrpA of C. pneumoniae, indicating that some of chlamydial surface molecules share antigenic epitopes. In contrast, the VD2, as well as the VD3, regions of the MOMP of C. pneumoniae were only recognized by C. pneumoniae-positive sera, suggesting the existence of species-specific epitopes. The identification of such epitopes of cell surface molecules provides new insights into C. pneumoniae-specific immune responses and may be of value for the improvement of C. pneumoniae-specific diagnostic assay systems based on defined recombinant antigens. PMID- 12734237 TI - Microbial population diversity in the urethras of healthy males and males suffering from nonchlamydial, nongonococcal urethritis. AB - Nonchlamydial, nongonococcal urethritis (NCNGU) is suggested to be a sexually transmitted disease in men. NCNGU patients were compared to control subjects with regard to the presence of potentially infectious bacteria in the first void urine. Patients' pre- and post-antibiotic-treatment urine samples and two samples obtained 2 weeks apart from healthy volunteers, who did not receive antibiotic therapy, were analyzed with broad-spectrum PCR tests aiming at eubacterial small subunit rRNA genes. Restriction fragment length polymorphism analysis of the amplicons cloned from the mixtures of PCR products revealed that many different species of microorganisms were found to be colonizing the male urethra. We document here clear differences in the composition of the resident urethral flora between samples obtained from various individuals and between samples obtained at various points in time for a single individual. No major changes in population complexity were found upon antimicrobial treatment. In two of five patients a previously suggested pathogen (Mycoplasma genitalium or Haemophilus parainfluenzae) was accurately identified on the basis of DNA sequencing. No ubiquitous, azithromycin-sensitive organism was identified as a common pathogen in all patients, but up to 40% of all clones represented as-yet-unclassified bacterial species. Relatively often Pseudomonas spp. or Pseudomonas-like organisms were identified in the bacterial flora of patients. Interestingly, an as-yet-uncharacterized microbial species was identified as a negative predictor of NCNGU. This species was identified in all control subjects and was absent from all of the patient' samples (5 of 5 versus 0 of 5, P = 0.0079). This suggests that NCNGU might also be diagnosed by assessing the absence rather than the presence of certain bacterial species. PMID- 12734235 TI - Snapshot of moving and expanding clones of Mycobacterium tuberculosis and their global distribution assessed by spoligotyping in an international study. AB - The present update on the global distribution of Mycobacterium tuberculosis complex spoligotypes provides both the octal and binary descriptions of the spoligotypes for M. tuberculosis complex, including Mycobacterium bovis, from >90 countries (13,008 patterns grouped into 813 shared types containing 11,708 isolates and 1,300 orphan patterns). A number of potential indices were developed to summarize the information on the biogeographical specificity of a given shared type, as well as its geographical spreading (matching code and spreading index, respectively). To facilitate the analysis of hundreds of spoligotypes each made up of a binary succession of 43 bits of information, a number of major and minor visual rules were also defined. A total of six major rules (A to F) with the precise description of the extra missing spacers (minor rules) were used to define 36 major clades (or families) of M. tuberculosis. Some major clades identified were the East African-Indian (EAI) clade, the Beijing clade, the Haarlem clade, the Latin American and Mediterranean (LAM) clade, the Central Asian (CAS) clade, a European clade of IS6110 low banders (X; highly prevalent in the United States and United Kingdom), and a widespread yet poorly defined clade (T). When the visual rules defined above were used for an automated labeling of the 813 shared types to define nine superfamilies of strains (Mycobacterium africanum, Beijing, M. bovis, EAI, CAS, T, Haarlem, X, and LAM), 96.9% of the shared types received a label, showing the potential for automated labeling of M. tuberculosis families in well-defined phylogeographical families. Intercontinental matches of shared types among eight continents and subcontinents (Africa, North America, Central America, South America, Europe, the Middle East and Central Asia, and the Far East) are analyzed and discussed. PMID- 12734238 TI - Controlled comparison of BACTEC 13A, MYCO/F LYTIC, BacT/ALERT MB, and ISOLATOR 10 systems for detection of mycobacteremia. AB - To compare the performance of the BACTEC 13A (Becton Dickinson, Sparks, Md.), BACTEC MYCO/F LYTIC (Becton Dickinson), BacT/ALERT MB (bioMerieux, Durham, N.C.), and ISOLATOR 10 lysis-centrifugation (Wampole Laboratories, Cranbury, N.J.) systems for detection of mycobacteremia in adults, we inoculated 5-ml aliquots of blood from patients with suspected mycobacteremia into the bottle or tube required for each system. Of 600 sets tested, 85 (14%) yielded Mycobacterium avium complex (MAC) and 9 (2%) yielded other species of mycobacteria. Of 26 complete (three bottles and one tube) adequately filled (5 +/- 1 ml) sets from which MAC was recovered, BACTEC 13A was positive for 19 (73%), BACTEC MYCO/F LYTIC was positive for 21 (81%), BacT/ALERT MB was positive for 22 (85%), and ISOLATOR 10 was positive for 21 (81%). Of the six possible two-way comparisons, the mean times to detection for the recovery of MAC from each bottle in positive adequately paired sets were 15.3 days for BACTEC 13A versus 12.8 days for MYCO/F LYTIC for 33 of 340 pairs, 14.1 days for BACTEC 13A versus 11.6 days for BacT/ALERT MB for 38 of 380 pairs, 12.6 days for BACTEC 13A versus 20.0 days for ISOLATOR 10 for 26 of 261 pairs, 12.8 days for BACTEC MYCO/F LYTIC versus 11.0 days for BacT/ALERT MB for 33 of 340 pairs, 13.2 days for BACTEC MYCO/F LYTIC versus 20.4 days for ISOLATOR 10 for 24 of 230 pairs, and 9.9 days for BacT/ALERT MB versus 19.0 days for ISOLATOR 10 for 24 of 257 pairs. There were no significant differences in yields between the systems. However, the mean time to detection differed significantly among the systems. The time to detection was shortest for BacT/ALERT MB, followed by BACTEC MYCO/F LYTIC and BACTEC 13A and then ISOLATOR 10. Although the numbers were too small for statistical comparison, the time to detection was substantially shorter for MAC than for Mycobacterium tuberculosis complex in the liquid systems. The continuously monitored systems (BACTEC MYCO/F LYTIC and BacT/ALERT MB) were as sensitive and, on balance, faster for the detection of MAC bacteremia than were the heretofore standard manual ISOLATOR 10 and radiometric BACTEC 13A systems. PMID- 12734236 TI - Improved detection of rhinoviruses by nucleic acid sequence-based amplification after nucleotide sequence determination of the 5' noncoding regions of additional rhinovirus strains. AB - The isothermal nucleic acid sequence-based amplification (NASBA) system was applied for the detection of rhinoviruses using primers targeted at the 5' noncoding region (5' NCR) of the viral genome. The nucleotide sequence of the 5' NCRs of 34 rhinovirus isolates was determined to map the most conserved regions and design more appropriate primers and probes. The assay amplified RNA extracted from 30 rhinovirus reference strains and 88 rhinovirus isolates, it did not amplify RNA from 49 enterovirus isolates and other respiratory viruses. The assay allows one to discriminate between group A and B rhinoviruses. Sensitivities for the detection of group B and group A rhinoviruses was 20 and 200 50% tissue culture infective doses, respectively. PMID- 12734239 TI - Simple and fast lateral flow test for classification of leprosy patients and identification of contacts with high risk of developing leprosy. AB - The interruption of leprosy transmission is one of the main challenges for leprosy control programs since no consistent evidence exists that transmission has been reduced after the introduction of multidrug therapy. Sources of infection are primarily people with high loads of bacteria with or without clinical signs of leprosy. The availability of a simple test system for the detection of antibodies to phenolic glycolipid-I (PGL-I) of Mycobacterium leprae to identify these individuals may be important in the prevention of transmission. We have developed a lateral flow assay, the ML Flow test, for the detection of antibodies to PGL-I which takes only 10 min to perform. An agreement of 91% was observed between enzyme-linked immunosorbent assay and our test; the agreement beyond chance (kappa value) was 0.77. We evaluated the use of whole blood by comparing 539 blood and serum samples from an area of high endemicity. The observed agreement was 85.9% (kappa = 0.70). Storage of the lateral flow test and the running buffer at 28 degrees C for up to 1 year did not influence the results of the assay. The sensitivity of the ML Flow test in correctly classifying MB patients was 97.4%. The specificity of the ML Flow test, based on the results of the control group, was 90.2%. The ML Flow test is a fast and easy-to-perform method for the detection of immunoglobulin M antibodies to PGL-I of M. leprae. It does not require any special equipment, and the highly stable reagents make the test robust and suitable for use in tropical countries. PMID- 12734240 TI - Usefulness of the MicroSeq 500 16S ribosomal DNA-based bacterial identification system for identification of clinically significant bacterial isolates with ambiguous biochemical profiles. AB - Due to the inadequate automation in the amplification and sequencing procedures, the use of 16S rRNA gene sequence-based methods in clinical microbiology laboratories is largely limited to identification of strains that are difficult to identify by phenotypic methods. In this study, using conventional full sequence 16S rRNA gene sequencing as the "gold standard," we evaluated the usefulness of the MicroSeq 500 16S ribosomal DNA (rDNA)-based bacterial identification system, which involves amplification and sequencing of the first 527-bp fragment of the 16S rRNA genes of bacterial strains and analysis of the sequences using the database of the system, for identification of clinically significant bacterial isolates with ambiguous biochemical profiles. Among 37 clinically significant bacterial strains that showed ambiguous biochemical profiles, representing 37 nonduplicating aerobic gram-positive and gram-negative, anaerobic, and Mycobacterium species, the MicroSeq 500 16S rDNA-based bacterial identification system was successful in identifying 30 (81.1%) of them. Five (13.5%) isolates were misidentified at the genus level (Granulicatella adiacens was misidentified as Abiotrophia defectiva, Helcococcus kunzii was misidentified as Clostridium hastiforme, Olsenella uli was misidentified as Atopobium rimae, Leptotrichia buccalis was misidentified as Fusobacterium mortiferum, and Bergeyella zoohelcum was misidentified as Rimerella anatipestifer), and two (5.4%) were misidentified at the species level (Actinomyces odontolyticus was misidentified as Actinomyces meyeri and Arcobacter cryaerophilus was misidentified as Arcobacter butzleri). When the same 527-bp DNA sequences of these seven isolates were compared to the known 16S rRNA gene sequences in the GenBank, five yielded the correct identity, with good discrimination between the best and second best match sequences, meaning that the reason for misidentification in these five isolates was due to a lack of the 16S rRNA gene sequences of these bacteria in the database of the MicroSeq 500 16S rDNA-based bacterial identification system. In conclusion, the MicroSeq 500 16S rDNA-based bacterial identification system is useful for identification of most clinically important bacterial strains with ambiguous biochemical profiles, but the database of the MicroSeq 500 16S rDNA-based bacterial identification system has to be expanded in order to encompass the rarely encountered bacterial species and achieve better accuracy in bacterial identification. PMID- 12734241 TI - Detection of anti-West Nile virus immunoglobulin M in chicken serum by an enzyme linked immunosorbent assay. AB - The emergence of West Nile (WN) virus in New York and the surrounding area in 1999 prompted an increase in surveillance measures throughout the United States, including the screening of sentinel chicken flocks for antibodies. An enzyme linked immunosorbent assay (ELISA) for the detection of chicken immunoglobulin M (IgM) to WN virus was developed, standardized, and characterized as a rapid and sensitive means to detect WN viral antibodies in sentinel flocks. Serum specimens from experimentally infected chickens were analyzed by using this assay, and IgM was detected as early as 3 to 7 days postinfection. Persistence of IgM varied from at least 19 to more than 61 days postinfection, which indicates the need to bleed sentinel flocks at least every 2 weeks for optimal results if this method is to be used as a screening tool. The ELISA was compared to hemagglutination inhibition and plaque reduction neutralization tests and was found to be the method of choice when early detection of WN antibody is required. House sparrows and rock doves are potential free-ranging sentinel species for WN virus, and the chicken WN IgM-capture ELISA was capable of detecting anti-WN IgM in house sparrow serum samples from laboratory-infected birds but not from rock dove serum samples. The chicken WN IgM-capture ELISA detected anti-WN antibodies in serum samples from naturally infected chickens. It also detected IgM in serum samples from two species of geese and from experimentally infected ring-necked pheasants, American crows, common grackles, and redwinged blackbirds. However, the test was determined to be less appropriate than an IgG (IgY)-based assay for use with free ranging birds. The positive-to-negative ratios in the ELISA were similar regardless of the strain of WN viral antigen used, and only minimal cross reactivity was observed between the WN and St. Louis encephalitis (SLE) IgM capture ELISAs. A blind-coded serum panel was tested, and the chicken WN IgM capture ELISA produced consistent results, with the exception of one borderline result. A preliminary test was done to assess the feasibility of a combined SLE and WN IgM-capture ELISA, and results were promising. PMID- 12734242 TI - DNA-level characterization of Helicobacter pylori strains from patients with overt disease and with benign infections in Bangladesh. AB - The complex relation between the genotype of Helicobacter pylori and its association with clinical outcome is not well understood. Studies in the West have showed that strains expressing certain virulence factors (vacAs1, vacAm1, and cagA) are associated with duodenal ulcer disease. However, the H. pylori genotype is known to vary with geographic region. In the present study, we compared several virulence markers (cagA, vacA, and iceA) and neutral markers (IS605, IS606, and IS608) in H. pylori strains isolated from 65 adult patients with peptic ulcer (PU) and 50 patients with nonulcer dyspepsia (NUD). PCR tests indicated that cagA is present in 75% of the strains from patients with PU compared to 55% in patients with NUD, and 80% of the isolates from patients with PU carried potentially toxigenic vacAs1 alleles of the vacuolating cytotoxin gene (vacA) compared to 60% in isolates from patients with NUD. However, no significant difference in any other virulence marker was observed in isolates from both groups. Phylogenetic analysis of the vacA middle region and the 5' end of the cagA gene indicates that Bangladeshi isolates are more closely related to H. pylori isolates from India and are different from isolates from East Asia. PMID- 12734244 TI - Two international methicillin-resistant Staphylococcus aureus clones endemic in a university hospital in Patras, Greece. AB - Pulsed-field gel electrophoresis (PFGE) of SmaI macrofragments and hybridization of ClaI digests with the mecA- and Tn554-specific DNA probes were used to define the endemic clones of methicillin-resistant Staphylococcus aureus (MRSA) among strains collected in 1993 and 1998 to 2000 at the University Hospital of Patras, Patras, Greece. Representatives of each clonal type were analyzed by spaA typing, multilocus sequence typing (MLST), and staphylococcal chromosomal cassette mec (SCCmec) typing. The results indicated the existence of two successive international MRSA clones: (i) a clonal type with PFGE type A, sequence type (ST) 30 (ST30), and SCCmec type IV, which was very similar to a clone widely spread in the United Kingdom, Mexico, and Finland, and (ii) a clonal type with PFGE type B, ST239, and SCCmec III, which was related to the Brazilian clone. Both clones seem to be widespread in Greece as well. A novel MRSA clone is also described and is characterized by a new MLST type (ST80) associated with SCCmec type IV and with the presence of Panton-Valentine leukocidin genes. PMID- 12734243 TI - Molecular epidemiology of Mycobacterium avium subsp. paratuberculosis: evidence for limited strain diversity, strain sharing, and identification of unique targets for diagnosis. AB - The objectives of this study were to understand the molecular diversity of animal and human strains of Mycobacterium avium subsp. paratuberculosis isolated in the United States and to identify M. avium subsp. paratuberculosis-specific diagnostic molecular markers to aid in disease detection, prevention, and control. Multiplex PCR of IS900 integration loci (MPIL) and amplified fragment length polymorphism (AFLP) analyses were used to fingerprint M. avium subsp. paratuberculosis isolates recovered from animals (n = 203) and patients with Crohn's disease (n = 7) from diverse geographic localities. Six hundred bacterial cultures, including M. avium subsp. paratuberculosis (n = 303), non-M. avium subsp. paratuberculosis mycobacteria (n = 129), and other nonmycobacterial species (n = 168), were analyzed to evaluate the specificity of two IS900 integration loci and a newly described M. avium subsp. paratuberculosis-specific sequence (locus 251) as potential targets for the diagnosis of M. avium subsp. paratuberculosis. MPIL fingerprint analysis revealed that 78% of bovine origin M. avium subsp. paratuberculosis isolates clustered together into a major node, whereas isolates from human and ovine sources showed greater genetic diversity. MPIL analysis also showed that the M. avium subsp. paratuberculosis isolates from ovine and bovine sources from the same state were more closely associated than were isolates from different geographic regions, suggesting that some of the strains are shared between these ruminant species. AFLP fingerprinting revealed a similar pattern, with most isolates from bovine sources clustering into two major nodes, while those recovered from sheep or humans were clustered on distinct branches. Overall, this study identified a high degree of genetic similarity between M. avium subsp. paratuberculosis strains recovered from cows regardless of geographic origin. Further, the results of our analyses reveal a relatively higher degree of genetic heterogeneity among M. avium subsp. paratuberculosis isolates recovered from human and ovine sources. PMID- 12734246 TI - Monitoring cytomegalovirus infection in adult and pediatric bone marrow transplant recipients by a real-time PCR assay performed with blood plasma. AB - The objective of this study was to evaluate the advantages of cytomegalovirus (CMV) real-time PCR in blood plasma to monitor CMV infection in a population of adult and pediatric bone marrow recipients in comparison with the pp65 antigenemia method. Fifty allogeneic bone marrow transplant recipients from our center, including 23 adults and 27 children, were enrolled. A CMV real-time PCR designed to amplify a well-conserved region of the UL123 gene was evaluated for its results with whole blood and blood plasma. The CMV real-time PCR assay and the CMV antigenemia method were performed in parallel with 558 blood samples. The results obtained by the two techniques were significantly correlated (r = 0.732; P < 0.0001). Twenty patients developed at least one episode of CMV replication, with a total of 24 episodes detected by CMV PCR; antigenemia assays were positive in 17 of these 24 episodes. The first positive PCR test preceded the first positive antigenemia by a median of 8 days. The median time interval necessary to obtain a negative CMV PCR test after implementation of preemptive treatment was 28 days. CMV PCR of plasma was positive in two children with CMV disease (one with early CMV pneumonia and one with CMV gastroenteritis), while CMV antigenemia remained negative. The use of CMV PCR with plasma to guide both implementation and discontinuation of CMV preemptive therapy might reduce the risk of occurrence of CMV disease since patients would be treated earlier, and it might also help to reduce the duration of treatment, which could attenuate the side effects of antiviral drugs. PMID- 12734245 TI - Characterization of atypical enteropathogenic Escherichia coli strains harboring the astA gene that were associated with a waterborne outbreak of diarrhea in Japan. AB - The virulence traits of the Escherichia coli strain associated with a waterborne diarrhea outbreak were examined. Forty-one of 75 students (ages 12 to 15) in Akita Prefecture, Japan, showed clinical symptoms. Seven E. coli Ouk:K-:H45 isolates were isolated from the patients as the causative agent of this outbreak. One isolate (EC-3605) showed the presence of E. coli attaching-and-effacing (eaeA) and enteroaggregative E. coli heat-stable enterotoxin-1 (astA) genes and the absence of Shiga toxin (stx1 and stx2) genes. A polymorphic enteropathogenic E. coli (EPEC) adherence factor plasmid was detected in EC-3605 with a major structural gene deletion and a regulatory gene frameshift mutation, revealing that EC-3605 represents an atypical EPEC strain harboring the astA gene. The role that atypical EPEC strains harboring the astA gene play in human disease is unclear. Our results, along with those of others, present a possibility that these strains comprise a distinct category of diarrheagenic E. coli and that astA affects the age distribution of atypical-EPEC infection. PMID- 12734248 TI - Applications of competitor RNA in diagnostic reverse transcription-PCR. AB - Detection of RNA viruses by reverse transcription (RT)-PCR has proven to be a useful approach for the diagnosis of infections caused by many viral pathogens. However, adequate controls are required for each step of the RT-PCR protocol to ensure the accuracies of diagnostic test results. Heterologous competitor RNA can be used as a control for a number of different aspects of diagnostic RT-PCR. Competitor RNA can be applied to assessments of the efficiency of RNA recovery during extraction procedures, detection of endogenous RT-PCR inhibitors that could lead to false-negative results, and quantification of viral template in samples used for diagnosis; competitor RNA can also be used as a positive control for the RT-PCR. In the present study, heterologous competitor RNA was synthesized by a method that uses two long oligonucleotide primers containing primer binding sites for RT-PCR amplification of porcine reproductive and respiratory syndrome virus or West Nile virus. Amplification of the competitor RNA by RT-PCR resulted in a product that was easily distinguished from the amplification product of viral RNA by agarose gel electrophoresis. Assessment of a variety of RNA samples prepared from routine submissions to a veterinary diagnostic laboratory found that either partial or complete inhibition of the RT-PCR could be demonstrated for approximately 20% of the samples. When inhibition was detected, either dilution of the sample or RNA extraction by an alternative protocol proved successful in eliminating the source of inhibition. PMID- 12734247 TI - Epidemiology of Cyclospora cayetanensis and other intestinal parasites in a community in Haiti. AB - We conducted an exploratory investigation in a community in Haiti to determine the prevalence of Cyclospora cayetanensis infection and to identify potential risk factors for C. cayetanensis infection. In 2001, two cross-sectional stool surveys and a nested case-control study were conducted. In 2002, a follow-up cross-sectional stool survey was conducted among children < or =10 years of age. Stool specimens from study participants and water samples from their wells were examined for Cyclospora and other intestinal parasites. In stools, the prevalence of infection with Cyclospora in persons of all ages decreased from 12% (20 of 167 persons) in February 2001 to 1.1% (4 of 352 persons) in April 2001, a 90.8% decrease. For children < or =10 years of age, the prevalence rates were 22.5% (16 of 71 children) in February 2001, 3.0% (4 of 135 children) in April 2001, and 2.5% (2 of 81 children) in January 2002. Use of the water from the artesian well in the northern region of the community versus the one in the south was the only risk factor associated with Cyclospora infection in multivariate analyses (odds ratio, 18.5; 95% confidence interval, 2.4 to 143.1). The water sample from one of the nine wells or water sources tested (one sample per source) in January 2001, shortly before the investigation began, was positive for Cyclospora by UV fluorescence microscopy and PCR. None of the water samples from the 46 wells or water sources tested during the investigation (one sample per source per testing period, including the artesian wells) were positive for Cyclospora. Further studies are needed to assess the role of water as a possible risk factor for Cyclospora infection in Haiti and other developing countries. PMID- 12734249 TI - Development and verification of an automated sample processing protocol for quantitation of human immunodeficiency virus type 1 RNA in plasma. AB - We developed and verified an automated sample processing protocol for use with the AMPLICOR HIV-1 MONITOR test, version 1.5 (Roche Diagnostics, Indianapolis, Ind.). The automated method uses the MagNA Pure LC instrument and total nucleic acid reagents (Roche Applied Science, Indianapolis, Ind.) to extract human immunodeficiency virus type 1 (HIV-1) RNA from plasma specimens. We compared the HIV-1 load results for a dilution series (1 to 5 nominal log(10) copies/ml) and 175 clinical specimens processed by the automated method to those for the same samples processed by the manual methods specified by the manufacturer. The sensitivity, dynamic range, and precision of the viral load assay obtained by automated processing of specimens were similar to those obtained by an ultrasensitive manual processing method. The results were highly correlated (R(2), 0.95), and were in close agreement, with a mean difference of 0.09 log(10) (standard deviation, 0.292). The limits of agreement were +/-0.58 log(10) for results for samples processed by both the manual and the automated methods. These performance characteristics were achieved with a smaller sample volume (200 versus 500 microl) and without a high-speed centrifugation step and required only 15 min of labor for a batch of 32 samples. In conclusion, the automated sample preparation protocol can replace both the standard and the ultrasensitive manual methods used with the AMPLICOR HIV-1 MONITOR test and can substantially reduce the labor associated with this test. PMID- 12734251 TI - Pgi genotyping is a surrogate for serotyping of encapsulated Haemophilus influenzae. AB - Study of the epidemiology of invasive infections caused by encapsulated Haemophilus influenzae has been complicated by the poor sensitivity and specificity of the serologic assays used to identify specific capsular polysaccharides. The population structure of these bacteria is highly clonal, however, and serotype is highly correlated with other genetic characteristics. We sought to determine if alleles of the highly conserved phosphoglucose isomerase (pgi) gene correspond to the serotypes of encapsulated H. influenzae strains. pgi alleles of 52 well-characterized encapsulated H. influenzae isolates were amplified by PCR, sequenced, and compared to one another and to additional previously reported H. influenzae pgi alleles. Overall, 83% of the strains possessed pgi alleles associated with the major serotype a, b, e, and f clonotypes that cause the most invasive disease in the United States. Six strains (four type a and two type f) had unusual pgi alleles, which suggested that these strains belonged to less common clonotypes of encapsulated bacteria or were actually nontypeable strains. pgi genotyping may provide a simple and stable surrogate for capsular serotyping. Further studies correlating pgi typing with the expression of capsule are likely to increase our understanding of the epidemiology and pathogenesis of these infections. PMID- 12734250 TI - Multilocus sequence typing and evolutionary relationships among the causative agents of melioidosis and glanders, Burkholderia pseudomallei and Burkholderia mallei. AB - A collection of 147 isolates of Burkholderia pseudomallei, B. mallei, and B. thailandensis was characterized by multilocus sequence typing (MLST). The 128 isolates of B. pseudomallei, the causative agent of melioidosis, were obtained from diverse geographic locations, from humans and animals with disease, and from the environment and were resolved into 71 sequence types. The utility of the MLST scheme for epidemiological investigations was established by analyzing isolates from captive marine mammals and birds and from humans in Hong Kong with melioidosis. MLST gave a level of resolution similar to that given by pulsed field gel electrophoresis and identified the same three clones causing disease in animals, each of which was also associated with disease in humans. The average divergence between the alleles of B. thailandensis and B. pseudomallei was 3.2%, and there was no sharing of alleles between these species. Trees constructed from differences in the allelic profiles of the isolates and from the concatenated sequences of the seven loci showed that the B. pseudomallei isolates formed a cluster of closely related lineages that were fully resolved from the cluster of B. thailandensis isolates, confirming their separate species status. However, isolates of B. mallei, the causative agent of glanders, recovered from three continents over a 30-year period had identical allelic profiles, and the B. mallei isolates clustered within the B. pseudomallei group of isolates. Alleles at six of the seven loci in B. mallei were also present within B. pseudomallei isolates, and B. mallei is a clone of B. pseudomallei that, on population genetics grounds, should not be given separate species status. PMID- 12734252 TI - Evaluation of a new serotyping assay for detection of anti-hepatitis C virus type specific antibodies in serum samples. AB - The performance of a new version (HC03) of the hepatitis C virus (HCV) serotyping 1-6 assay (Abbott Murex Laboratories), a specific test for serological determination of HCV types, was evaluated using a selected panel of 180 HCV RNA positive sera. HC03 was more sensitive than the current HC02 version, typing 53 (37.6%) of 141 samples which were not typable with HC02. Furthermore, the HC03 specificity was 94.1% as evaluated with a panel of 22 genotyped samples. This new version of the test improves the quality of the serological approach to HCV type determination. PMID- 12734253 TI - Genetic characterization of a novel, naturally occurring recombinant human G6P[6] rotavirus. AB - A binary classification system has been established for group A rotaviruses, with the viral capsid protein VP7 defining G types and VP4 defining P types. At least 15 G types and 21 P types have been isolated globally with various G and P combinations. Most of the currently circulating human rotaviruses belong to G1P[8], G2P[4], G3P[8], and G4P[8]. We report a human rotavirus strain (B1711) with a novel genotypic VP7/VP4 combination of G6P[6]. This unique rotavirus was isolated from a 13-month-old human immunodeficiency virus (HIV)- negative child of an HIV-seropositive Malian mother that was hospitalized with severe diarrhea in Belgium after returning from a trip to Mali. The VP7 and VP4 genes of the rotavirus strain were sequenced, and phylogenetic trees were constructed. Nucleotide and amino acid sequence comparisons with 15 known G genotypes indicated that the VP7 sequence of strain B1711 was most closely related to an American (Se584) and an Italian (PA151) human G6 strain (95 to 96% nucleotide and 98% amino acid identity). Comparison of the VP4 sequence with 21 P types showed the closest similarity to P[6] genotypes, with greatest similarity to a G8P[6] Malawi strain (mw131) (97% nucleotide and 98% amino acid identity). The B1711 strain is the first reported rotavirus isolate with a G6P[6] genotypic combination. The discovery and surveillance of novel human and nonhuman rotavirus G or P types or of novel G/P combinations is essential for the design of future rotavirus vaccines and for our understanding of rotavirus diversity and evolution. PMID- 12734254 TI - Evaluation of the Vitek 2 ID-GNB assay for identification of members of the family Enterobacteriaceae and other nonenteric gram-negative bacilli and comparison with the Vitek GNI+ card. AB - We evaluated the Vitek 2 ID-GNB identification card (bioMerieux, Inc., Durham, N.C.) for its ability to identify members of the family Enterobacteriaceae and other gram-negative bacilli that are isolated in clinical microbiology laboratories. Using 482 enteric stock cultures and 103 strains of oxidase positive, gram-negative glucose-fermenting and nonfermenting bacilli that were maintained at -70 degrees C and passaged three times before use, we inoculated cards according to the manufacturer's directions and processed them in a Vitek 2 instrument using version VT2-R02.03 software. All panel identifications were compared to reference identifications previously confirmed by conventional tube biochemical assays. At the end of the initial 3-h incubation period, the Vitek 2 instrument demonstrated an accuracy of 93.0% for the identification of enteric strains; 414 (85.9%) were correctly identified at probability levels ranging from excellent to good, and an additional 34 (7.1%) strains were correctly identified but at a low level of discrimination. Nineteen (3.9%) strains were unidentified, and 15 (3.1%) were misidentified. The 19 unidentified strains were scattered among 10 genera. Three of the 15 misidentified strains were lactose-positive Salmonella spp. and were identified as Escherichia coli; another was a lactose positive, malonate-negative Salmonella enterica subsp. arizonae strain that was identified as E. coli. Of the 103 glucose-fermenting and nonfermenting nonenteric strains, 88 (85.4%) were correctly identified at probability levels ranging from excellent to good, and 10 (9.7%) were correctly identified, but at a low level of discrimination, for a total of 95.1% accuracy with this group. Two strains were unidentified and three were misidentified. The errors occurred for strains in three different genera. With the increased hands-off approach of the Vitek 2 instrument and accuracies of 93% for the identification of enteric organisms and 95.1% for the identification of nonenteric organisms with the ID-GNB card, use of this product presents an acceptable method for the identification of most gram negative organisms commonly isolated in the clinical laboratory. A comparison of these results to those obtained by testing 454 of the same strains with the Vitek GNI+ card revealed no significant difference in the abilities of the two cards to identify these organisms accurately. PMID- 12734255 TI - Recombinant VP7-based enzyme-linked immunosorbent assay for detection of immunoglobulin G antibodies to Colorado tick fever virus. AB - VP6, VP7, VP9, VP10, VP11, and VP12 of Colorado tick fever virus (CTF virus), a virus member of the genus Coltivirus, family Reoviridae, were expressed in bacteria with the pGEX-4T-2 vector. A partial sequence of VP7 (designated pVP7) was chosen to elaborate an enzyme-linked immunosorbent assay (ELISA) for detecting anti-CTF virus immunoglobulin G (IgG) antibodies in humans. This was based on two observations: (i) among all expressed proteins, pVP7 showed the highest immunoreactivity to an anti-CTF virus hyperimmune ascitic fluid; (ii) to provide the highest selectivity of antibody detection, the expressed sequence was chosen within a region which is highly divergent (49% amino acid identity) from the homologous sequence of another coltivirus, the Eyach virus. The pVP7 ELISA was evaluated with 368 serum samples from French blood donors and found to provide 98.1% specificity. Assays with the Calisher set of human serum samples, positive for anti-CTF virus antibodies (C. H. Calisher, J. D. Poland, S. B. Calisher, and L. A Warmoth, J. Clin. Microbiol. 22:84-88, 1985), showed that the pVP7 ELISA provided 100% sensitivity for the tested population. After elaboration of recombinant-protein-based ELISAs for diagnosis of infections with members of the viral genera Orbivirus, Orthoreovirus, and Rotavirus, it was shown that a recombinant protein could be used to detect antibodies to the human pathogen Colorado tick fever virus. PMID- 12734256 TI - Identification and characterization of a new variant of Shiga toxin 1 in Escherichia coli ONT:H19 of bovine origin. AB - A new variant of Shiga toxin 1 (Stx1), designated Stx1d, which deviates considerably more than any other known variant from Stx1 encoded by phage 933J, was identified in an Escherichia coli strain, ONT:H19, isolated from bovine feces. The complete stx(1) gene of this strain was amplified and sequenced. Nucleotide sequence homology with stx(1) from phage 933J was only 91%, resulting in the substitution of 20 amino acids in the A subunit and 7 amino acids in the B subunit of the protein. Cell culture supernatant of this strain, which was negative for stx(2) by PCR testing, was cytotoxic to Vero cells and gave positive results in two commercial enzyme-linked immunosorbent assays for Stx. PCR primers were constructed for the specific detection of the new variant. The findings of this study suggest that Stx1 is not as conserved as thought before and that there might be more variants which cannot be detected by commonly used PCR methods. PMID- 12734258 TI - Real-time PCR assay of individual human immunodeficiency virus type 1 variants in coinfected human lymphoid tissues. AB - In the course of infection, human immunodeficiency virus type 1 (HIV-1) mutates, diverging into a "swarm" of viral quasispecies, and the predominance of CCR5- or CXCR4-utilizing quasispecies is strongly associated with the pattern of disease progression. Quantification of CCR5- and CXCR4-utilizing viruses in viral swarms is important in the investigation of the mechanisms of this phenomenon. Here, we report on a new real-time PCR-based methodology for the evaluation of replication of individual CCR5- and CXCR4-utilizing variants. The assay is highly reproducible, with a coefficient of variation of <3%, and it accurately estimates the numbers of virus-specific RNA copies even when their difference in the mixture is 2 orders of magnitude. We demonstrate that replications of CCR5- and CXCR4-utilizing variants can be evaluated and distinguished in experimentally coinfected human lymphoid tissue. The assay we developed may facilitate study of the mechanisms of the R5-to-X4 switch in viral swarms in human tissues infected with HIV-1. PMID- 12734257 TI - Rapid identification of Escherichia coli pathotypes by virulence gene detection with DNA microarrays. AB - One approach to the accurate determination of the pathogenic potential (pathotype) of isolated Escherichia coli strains would be through a complete assessment of each strain for the presence of all known E. coli virulence factors. To accomplish this, an E. coli virulence factor DNA microarray composed of 105 DNA PCR amplicons printed on glass slides and arranged in eight subarrays corresponding to different E. coli pathotypes was developed. Fluorescently labeled genomic DNAs from E. coli strains representing known pathotypes were initially hybridized to the virulence gene microarrays for both chip optimization and validation. Hybridization pattern analysis with clinical isolates permitted a rapid assessment of their virulence attributes and determination of the pathogenic group to which they belonged. Virulence factors belonging to two different pathotypes were detected in one human E. coli isolate (strain H87 5406). The microarray was also tested for its ability to distinguish among phylogenetic groups of genes by using gene probes derived from the attaching-and effacing locus (espA, espB, tir). After hybridization with these probes, we were able to distinguish E. coli strains harboring espA, espB, and tir sequences closely related to the gene sequences of an enterohemorrhagic strain (EDL933), a human enteropathogenic strain (E2348/69), or an animal enteropathogenic strain (RDEC-1). Our results show that the virulence factor microarray is a powerful tool for diagnosis-based studies and that the concept is useful for both gene quantitation and subtyping. Additionally, the multitude of virulence genes present on the microarray should greatly facilitate the detection of virulence genes acquired by horizontal transfer and the identification of emerging pathotypes. PMID- 12734259 TI - Comparison of lateral-flow immunoassay and enzyme immunoassay with viral culture for rapid detection of influenza virus in nasal wash specimens from children. AB - The performance of two commercially available rapid test kits for influenza virus detection was compared to that of viral culture by using 356 nasal wash specimens collected during the 2001 to 2002 influenza season. Overall, the two rapid tests were easy to perform and showed comparable sensitivities (70.4 and 72.2%) and specificities (97.7 and 98.3%); for both test kit groups, most of the specimens that yielded false-negative results were found to be growing influenza B virus. PMID- 12734260 TI - Failure to genotype herpes simplex virus by real-time PCR assay and melting curve analysis due to sequence variation within probe binding sites. AB - Real-time PCR with melting curve analysis of PCR products is a rapid procedure for detecting and genotyping herpes simplex virus (HSV). When testing mucocutaneous samples for HSV by a real-time PCR assay targeting the DNA polymerase gene, we found that some PCR products had atypical melting curves that did not conform to the expected melting temperatures for HSV type 1 or 2. Sequence analysis showed that these strains had base-pair mismatches over the probe binding sites. An alternative assay is required to type such atypical isolates. PMID- 12734262 TI - Detection of fluconazole-resistant isolates of Candida glabrata by using an agar screen assay. AB - The ability of a fluconazole-containing agar screen assay to accurately detect isolates of Candida glabrata resistant to the azole antifungal agent fluconazole was evaluated on a collection of 100 clinical isolates of this organism. Results were correlated with the MIC of fluconazole for these isolates and compared with the results of a previously published disk diffusion-based fluconazole resistance screening test. Agar screen assay results were in categorical agreement with MIC based determinations for 97% (97/100) of the isolates tested. This correlation was higher than that obtained with the disk diffusion technique, which categorized only 87% (87/100) of isolates correctly, and suggests that the agar screening approach can effectively expedite fluconazole susceptibility testing of C. glabrata isolates. PMID- 12734261 TI - Multiplex PCR for detection of three plasmid-borne genes of enteroaggregative Escherichia coli strains. AB - We developed a novel multiplex PCR assay for enteroaggregative Escherichia coli (EAEC) detection, by using three plasmid-borne genes (the aggregative adherence [AA] probe, aap, and aggR). One or more of the loci were detected in 24 (86%) of 28 patient isolates analyzed. The multiplex PCR assay is a fast, convenient, and sensitive molecular test to detect EAEC. PMID- 12734263 TI - Kinetics and significance of serum hepatitis C virus core antigen in patients with acute hepatitis C. AB - An immunoassay detecting hepatitis C virus core antigen was evaluated for its ability to predict clinical outcome in a series of patients with acute hepatitis C. In subjects who cleared the virus, core antigen was no longer detectable within 16 weeks of onset, whereas considerable fluctuations were noted among patients progressing to chronic hepatitis, one of whom showed consistently negative values despite the intermittent presence of viral RNA. PMID- 12734264 TI - Characterization of Mycobacterium montefiorense sp. nov., a novel pathogenic Mycobacterium from moray eels that is related to Mycobacterium triplex. AB - The characterization of a novel Mycobacterium sp. isolated from granulomatous skin lesions of moray eels is reported. Analysis of the hsp65 gene, small-subunit rRNA gene, rRNA spacer region, and phenotypic characteristics demonstrate that this organism is distinct from its closest genetic match, Mycobacterium triplex, and it has been named M. montefiorense sp. nov. PMID- 12734265 TI - Performance of the OraQuick rapid antibody test for diagnosis of human immunodeficiency virus type 1 infection in patients with various levels of exposure to highly active antiretroviral therapy. AB - With oral mucosal transudate and serum samples from 101 human immunodeficiency virus type 1 (HIV-1)-infected subjects and 100 HIV-1-negative volunteers, the OraQuick HIV-1 test demonstrated 100% specificity and 96% sensitivity. Four false negative subjects, who were characterized by early initiation of effective antiretroviral therapy, demonstrated waning serum anti-gp41 titers and Western blot band intensities. PMID- 12734266 TI - Risk of occupational human herpesvirus 8 infection for health care workers. AB - Human herpesvirus 8 (HHV-8), or Kaposi's sarcoma-associated herpesvirus, is highly prevalent in certain risk groups (human immunodeficiency virus-infected patients, transplant recipients, and patients on hemodialysis). Heath care workers caring for these patients were found to be more frequently infected with HHV-8 than staff caring for other patients (P < 0.01). PMID- 12734267 TI - O-antigen serotypes and type III secretory toxins in clinical isolates of Pseudomonas aeruginosa. AB - The association of O-antigen serotypes with type III secretory toxins was analyzed in 99 clinical isolates of Pseudomonas aeruginosa. Isolates secreting ExoU were frequently serotyped as O11, but none were serotype O1. Most of the isolates that were nontypeable for O antigen did not secrete type III secretory toxins. PMID- 12734269 TI - Frequency of amantadine-resistant influenza A viruses during two seasons featuring cocirculation of H1N1 and H3N2. AB - In two influenza seasons during which H1N1 and H3N2 cocirculated, resistance was more frequent in H3N2 strains than in H1N1 strains after amantadine treatment. Predominant amino acid substitutions in M2 protein occurred at position 31 (serine to asparagine) in H3N2 strains and at position 27 (valine to alanine) in H1N1 strains. PMID- 12734268 TI - Usefulness of urinary antigen detection by an immunochromatographic test for diagnosis of pneumococcal pneumonia in children. AB - We evaluated an immunochromatographic assay detecting pneumococcal antigen in urine samples from children diagnosed with pneumococcal pneumonia. The sensitivity and specificity of the immunochromatographic test with nonconcentrated urine (NCU) were 86.7 and 62.9%, respectively; with concentrated urine (CU), they were 100 and 11.7%, respectively. Pneumococcal antigen was also detected in 42.5% of NCU and 87.1% of CU samples from nasopharyngeal carriers. This is a nonspecific test for the diagnosis of pneumococcal pneumonia in children, particularly the very young. PMID- 12734271 TI - Rapid screening for Streptococcus agalactiae in vaginal specimens of pregnant women by fluorescent in situ hybridization. AB - Group B streptococci (GBS) are the most frequent pathogens in neonates with sepsis. A rapid screening method is required to identify carriage of GBS in pregnant women at the time of delivery. In order to detect GBS in vaginal specimens, the efficiency of the standard culture versus fluorescent in situ hybridization (FISH) was investigated. In 258 examined vaginal specimens, FISH identified 58 of the 59 GBS-positive samples (98.3%), whereas by means of standard culture only 38 specimens were positive (64.4%). We recommend FISH as a rapid, specific, highly sensitive screening technique for the detection of GBS in pregnant women at delivery. PMID- 12734270 TI - A serotype V clone is predominant among erythromycin-resistant Streptococcus agalactiae isolates in a southwestern region of Germany. AB - One hundred ninety-three Streptococcus agalactiae isolates of neonatal origin and 146 isolates from adult women were analyzed for macrolide resistance and investigated for clonality. Among erythromycin-resistant isolates, serotype V turned out to be the most frequent. Comparative pulsed-field gel electrophoresis analysis revealed genetic clustering of resistant strains and predominance of a single clone family within an otherwise heterogeneous serotype V population. PMID- 12734272 TI - Pilot study of COBAS PCR and ligase chain reaction for detection of rectal infections due to Chlamydia trachomatis. AB - We tested rectal specimens from men who have sex with men for Chlamydia trachomatis by using COBAS PCR (Roche Diagnostics) and ligase chain reaction LCR (Abbott laboratories) and compared three PCR specimen-processing procedures. Chlamydiae were detected by one or more procedures in 22 of 186 specimens. All three PCR tests were positive for 17 specimens, all of which also tested positive by LCR. PMID- 12734273 TI - Variation in susceptibility of bloodstream isolates of Candida glabrata to fluconazole according to patient age and geographic location. AB - We examined the susceptibilities to fluconazole of 559 bloodstream infection isolates of Candida glabrata and grouped the isolates by patient age and geographic location within the United States. Susceptibility of C. glabrata to fluconazole was lowest in the Pacific (44%) and East South Central (47%) regions and was highest in the West South Central region (82%) (regions are as designated by the U.S. Bureau of the Census). Isolates from pediatric patients were virtually all susceptible to fluconazole, whereas the highest frequency of resistance was observed in isolates from patients 16 to 64 years of age. PMID- 12734274 TI - Comparison of the Denka-Seiken INFLU A.B-Quick and BD Directigen Flu A+B kits with direct fluorescent-antibody staining and shell vial culture methods for rapid detection of influenza viruses. AB - The INFLU A.B-Quick and Directigen Flu A+B enzyme immunoassays were compared with direct immunofluorescence and cell culture for detection of influenza A and B viruses in a total of 255 patient specimens. Both assays identified 23 of 42 influenza A viruses (sensitivity, 54.8%; specificity, 100%; positive predictive value [PPV], 100%; negative predictive value [NPV], 91.8%). The INFLU A.B-Quick assay identified 10 of 16 influenza B viruses (sensitivity, 62.5%; specificity, 99.6%; PPV, 90.9%; NPV, 97.5%), and the Directigen Flu A+B assay detected 9 of 16 influenza B viruses (sensitivity, 56.3%; specificity, 99.6%; PPV, 90%; NPV, 97.1%). PMID- 12734275 TI - Detection of circulating Aspergillus fumigatus galactomannan: value and limits of the Platelia test for diagnosing invasive aspergillosis. AB - The effectiveness of galactomannan detection with the Platelia test was evaluated in a prospective study of 3,327 sera from 807 patients. The specificity was 99.6% (748 of 751 cases). For the groups of patients with proven and probable invasive aspergillosis, the sensitivity was 50.0% (17 of 34 cases). The disappointing sensitivity associated with the presence of rare false-positive cases underlines the limits of this test. PMID- 12734276 TI - Human group B rotavirus infections cause severe diarrhea in children and adults in Bangladesh. AB - Human group B rotavirus was detected in 12 of 220 adult patients and 2 of 67 child patients with severe diarrhea in Bangladesh. Group B rotavirus may be virulent in both adults and children, and the virus may be an especially serious diarrheal agent in Bangladesh. PMID- 12734279 TI - Influence of lack of CO2 on anaerobic bacteria: a quick method to verify the absence of CO2. AB - Since CO(2) is carbonic acid gas, it will decrease the pH if added to water. Therefore, a simple test can determine whether CO(2) is present in a gas mixture. If the pH of gassed water does not change significantly, then CO(2) is not present in the gas. PMID- 12734278 TI - Extended-spectrum beta-lactamase enzymes in clinical isolates of Enterobacter species from Lagos, Nigeria. AB - Over a 9-month period, 8 of 40 nonduplicate isolates of Enterobacter spp. producing extended-spectrum beta-lactamase (ESBL) were detected for the first time from two hospitals in Lagos, Nigeria. Microbiologic and molecular analysis confirmed the presence of ESBL. Only four isolates transferred ESBL resistance as determined by the conjugation test, and pulsed-field gel electrophoresis showed genetically unrelated isolates. PMID- 12734277 TI - Multilocus sequence typing has better discriminatory ability for typing Vibrio cholerae than does pulsed-field gel electrophoresis and provides a measure of phylogenetic relatedness. AB - Twenty-two Vibrio cholerae isolates, including some from "epidemic" (O1 and O139) and "nonepidemic" serogroups, were characterized by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) by using three housekeeping genes, gyrB, pgm, and recA; sequence data were also obtained for the virulence-associated genes tcpA, ctxA, and ctxB. Even with the small number of loci used, MLST had better discriminatory ability than did PFGE. On MLST analysis, there was clear clustering of epidemic serogroups; much greater diversity was seen among tcpA- and ctxAB-positive V. cholerae strains from other, nonepidemic serogroups, with a number of tcpA and ctxAB alleles identified. PMID- 12734280 TI - Genotyping of Candida albicans oral strains from healthy individuals by polymorphic microsatellite locus analysis. AB - Analysis of a polymorphic microsatellite locus was applied to 85 Candida albicans strains from healthy individuals. Comparison with strains from nonhealthy individuals previously analyzed in our laboratory showed an overall similarity, suggesting that all commensal strains have the ability to develop as pathogens. PMID- 12734281 TI - Search for cytolethal distending toxin production among fecal Escherichia coli isolates from Brazilian children with diarrhea and without diarrhea. AB - The enteropathogenic role of cytolethal distending toxin-producing Escherichia coli was investigated by searching sequences homologous to the cdt genes of an O86 strain among 2,074 isolates from 200 children with acute diarrhea and 200 controls in Brazil. Only one (0.5%) diarrheic child and two (1.0%) non-diarrheic controls harbored cdt-positive isolates. PMID- 12734282 TI - Mutations in the rpoB gene of multidrug-resistant Mycobacterium tuberculosis isolates from China. AB - Mutations in the 81-bp rifampin resistance determining region (RRDR) and mutation V176F locating at the beginning of the ropB gene were analyzed by DNA sequencing of 86 Mycobacterium tuberculosis clinical isolates (72 resistant and 14 sensitive) from different parts of China. Sixty-five mutations of 22 distinct kinds, 21 point mutations, and 1 insertion were found in 65 of 72 resistant isolates. The most common mutations were in codons 531 (41%), 526 (40%), and 516 (4%). Mutations were not found in seven (10%) of the resistant isolates. Six new alleles within the RRDR, along with five novel mutations outside the RRDR, are reported. None of isolates contained the V176 mutation. PMID- 12734283 TI - Novel polymorphic region of the rpoB gene containing Mycobacterium species specific sequences and its use in identification of mycobacteria. AB - Sequence analysis of a specific region of the mycobacterium rpoB gene in 35 mycobacterial strains representing 26 different mycobacterial species of clinical importance showed that there exists a highly polymorphic region. Based on the sequences of the polymorphic region, the oligonucleotide probes of 14 mycobacterial species with relatively high clinical importance were designed and shown to be specific to their corresponding mycobacterial species by dot blot hybridization. The results showed that the probes designed in this study are highly specific to each mycobacterial species, which suggests that these sequences may be useful for the species identification of mycobacteria. PMID- 12734284 TI - Subcutaneous phaeohyphomycosis caused by Veronaea botryosa in the People's Republic of China. AB - The second case of phaeohyphomycosis caused by Veronaea botryosa in China, in a 12-year-old boy from Jiangsu Province, is presented. Based on direct examination of the scrapings from crusted lesions; histologic examination of the biopsy tissue showing septate, phaeoid hyphal elements; and the culture exhibiting sympodial, conidiogenous cells producing predominantly two-celled, cylindric conidia, the etiologic agent was identified as V. botryosa. PMID- 12734285 TI - Spondylodiscitis due to Clostridium ramosum infection in an immunocompetent elderly patient. AB - The first ever case of spondylodiscitis caused by Clostridium ramosum in an elderly immunocompetent patient has been reported. C. ramosum is usually an intestinal bacterium but may occasionally be isolated in clinical specimens as an opportunistic pathogen. This report shows that this anaerobic organism can cause bone tropism without there having been any contamination due to spinal surgery. The infection cleared after empirical therapy using intravenous amoxicillin and oral metronidazole. PMID- 12734286 TI - Neonatal sudden death due to Legionella pneumonia associated with water birth in a domestic spa bath. AB - We report the first case of neonatal Legionnaires' disease associated with water birth in a spa bath at home. Legionella pneumophila serogroup 6 was detected from postmortem lung tissue. PMID- 12734287 TI - Acute flaccid paralysis from echovirus type 33 infection. AB - During a community echovirus type 33 outbreak, the virus was detected in the feces and cerebrospinal fluid of a 3-year-old boy with right arm weakness that followed a mild nonspecific febrile illness. This is the first time an association between echovirus type 33 infection and acute flaccid paralysis has been reported. PMID- 12734288 TI - Infection with an extended-spectrum beta-lactamase-producing strain of Serratia marcescens following tongue reconstruction. AB - We report a case of postsurgical wound infection of polymicrobial etiology caused by Serratia marcescens and Pseudomonas aeruginosa following the use of a radial forearm free flap for oncological tongue reconstruction. S. marcescens was a producer of SHV-12 extended-spectrum beta-lactamase (ESBL). This is the first report from India of this ESBL. S. marcescens and P. aeruginosa were resistant to the empirical perioperative antibiotics administered. Delay in the recognition of the type of infection and in the institution of appropriate therapy resulted in total loss of the free flap. PMID- 12734289 TI - Eubacterium callanderi bacteremia: report of the first case. AB - Eubacterium callanderi is an environmental anaerobic rod-shaped bacterium first isolated in 1998 from an industrial anaerobic digester. We report on the first clinical isolate of E. callanderi, which was recovered from the blood of a patient with a bladder carcinoma. Identification of the organism was made by cell fatty acid chromatographic analysis and 16S rRNA gene sequencing. PMID- 12734290 TI - Fatal case of endocarditis due to Weissella confusa. AB - This is the first reported case of endocarditis due to the Lactobacillus-like vancomycin-resistant gram-positive bacillus Weissella confusa. Full identification and susceptibility testing of Lactobacillus-like organisms recovered in blood culture should be performed for patients with clinical presentations that suggest endocarditis. PMID- 12734291 TI - Soft tissue infection and bacteremia caused by Shewanella putrefaciens. AB - Shewanella putrefaciens is as yet rarely responsible for clinical syndromes in humans. However, a case involving multiple organs in an elderly male under treatment with appropriate steroids confirms that attention should be devoted to unusual pathogens. PMID- 12734293 TI - Bacteremia due to three Bacillus species in a case of Munchausen's syndrome. AB - We report on a case of recurrent bacteremia due to three Bacillus spp. in an immunocompetent patient with no history of intravenous drug use. The source of the organisms was postulated to be a self-injected compound containing Bacillus spores, given the patient's past history of psychiatric illness and self destructive behavior. PMID- 12734292 TI - Bacteremia due to a novel Microbacterium species in a patient with leukemia and description of Microbacterium paraoxydans sp. nov. AB - A yellow-pigmented coryneform rod was isolated from the blood of a child with acute lymphoblastic leukemia who was perfused with a central venous catheter. The culture bottles were positive twice, at a 2-month interval. The isolate was identified as a Microbacterium sp. and studied along with five other similar strains. Phenotypic, chemotaxonomic, and genetic characteristics indicated that they are closely related to Microbacterium oxydans but that they belong to a distinct species, for which the name Microbacterium paraoxydans sp. nov. is proposed. The type strain of M. paraoxydans is CF36(T) = DSM 15019(T). The G+C content of its DNA is 69.9 mol%. PMID- 12734294 TI - Babesia bicornis sp. nov. and Theileria bicornis sp. nov.: tick-borne parasites associated with mortality in the black rhinoceros (Diceros bicornis). AB - A novel Babesia species, designated Babesia bicornis sp. nov., was identified in three black rhinoceroses (Diceros bicornis) that died in wildlife areas in Tanzania and South Africa. Screening of black rhinoceroses in South Africa revealed, in addition to B. bicornis, a second parasite, designated Theileria bicornis sp. nov. PMID- 12734295 TI - Sepsis, multiple organ failure, and death due to Pandoraea pnomenusa infection after lung transplantation. AB - A 30-year-old man died with Pandoraea pnomenusa sepsis after lung transplantation. Pandoraea species are gram-negative rods, closely related to, and commonly misidentified as, Burkholderia cepacia complex or Ralstonia species. Heretofore considered soil bacteria and colonizers that infect patients with chronic lung diseases, Pandoraea species can produce severe infections. PMID- 12734296 TI - Peripartum transmission of penicillin-resistant Streptococcus pneumoniae. AB - Streptococcus pneumoniae is a rarely recognized cause of neonatal sepsis. We present a recent case of S. pneumoniae bacteremia acquired on the first day of life in a neonate born at 30 weeks of gestation to a mother without prenatal care who had prolonged rupture of the membranes and received intravenous ampicillin prior to delivery. The isolate was resistant to penicillin, with a MIC of the drug of 4 microg/ml. The child responded to a 7-day course of intravenous vancomycin. S. pneumoniae was recovered from the vagina of the mother on a swab culture collected prior to delivery, and isolates from mother and child were confirmed to be identical on the basis of pulsed-field gel electrophoresis. Although neonatal sepsis due to the peripartum transmission of S. pneumoniae is rare, this case highlights the concern that increasing efforts to prevent group B streptococcus neonatal disease may lead to an increase in neonatal infections due to resistant organisms. PMID- 12734297 TI - Keratitis caused by Scedosporium apiospermum successfully treated with a cornea transplant and voriconazole. AB - A case of Scedosporium apiospermum keratitis was successfully treated with oral voriconazole and penetrating keratoplasty. Voriconazole levels in the aqueous humor were 53% of the levels in plasma and exceeded the MIC for the isolate by sevenfold. PMID- 12734298 TI - Detection of Legionella pneumophila serogroup 1 antigen in bronchoalveolar lavage fluid by an immunochromatographic assay. PMID- 12734299 TI - Evidence for spread of a clonal strain of Pseudomonas aeruginosa among cystic fibrosis clinics. PMID- 12734300 TI - Clinical microbiology in the year 2025: serologic and host-oriented diagnosis. PMID- 12734301 TI - Criteria for identification of cross-contamination of cultures of Mycobacterium tuberculosis in routine microbiology laboratories. PMID- 12734302 TI - Immunochromatographic test for rapid detection of Streptococcus pneumoniae in the nasopharynx. PMID- 12734303 TI - Contamination management of broad-range or specific PCR: is there any difference? PMID- 12734304 TI - Judging new markers by their ability to improve predictive accuracy. PMID- 12734305 TI - Growing pains: central review board project still developing. PMID- 12734307 TI - Recent studies attempt to clarify relationship between oral cancer and human papillomavirus. PMID- 12734308 TI - Stat bite: Incidence of and mortality from cancer of the oral cavity and pharynx. PMID- 12734309 TI - Consortia, 'big science' part of a paradigm shift for genetic epidemiology. PMID- 12734310 TI - Report outlines plan to reform cancer care in Australia. PMID- 12734311 TI - Recommendations for Optimizing Cancer Care in Australia. PMID- 12734312 TI - Prevention methods underused, report concludes. PMID- 12734315 TI - Development of investigational radiation modifiers. PMID- 12734316 TI - Preclinical characterization of the antiglioma activity of a tropism-enhanced adenovirus targeted to the retinoblastoma pathway. AB - BACKGROUND: Oncolytic adenoviruses are promising therapies for the treatment of gliomas. However, untargeted viral replication and the paucity of coxsackie adenovirus receptors (CARs) on tumor cells are major stumbling blocks for adenovirus-based treatment. We studied the antiglioma activity of the tumor selective Delta-24 adenovirus, which encompasses an early 1 A adenoviral (E1A) deletion in the retinoblastoma (Rb) protein-binding region, and of the Delta-24 RGD adenovirus. Delta-24-RGD has an RGD-4C peptide motif inserted into the adenoviral fiber, which allows the adenovirus to anchor directly to integrins. METHODS: CAR and integrin expression were examined by flow cytometry in six glioma cell lines and in normal human astrocytes (NHAs). Adenoviral vectors containing green fluorescent protein (GFP) (AdGFP and AdGFP-RGD) were used to infect glioma cell lines with high or low CAR expression. Viability of glioma cells infected with different adenoviruses was assessed by trypan blue staining. Adenovirus replication was quantified with the infection-dose replication assay. Athymic mice carrying glioma xenografts received intratumoral injections of Delta 24-RGD or Delta-24 and were followed for survival, which was analyzed by the Kaplan-Meier method and the log-rank test. All statistical tests were two-sided. RESULTS: Half the glioma cell lines expressed low levels of CAR (defined as <50% of cells expressing detectable CAR); all lines expressed integrins in more than 50% of cells. Infection of U-87 MG cells (a low-CAR-expressing line) with AdGFP RGD resulted in approximately six times more GFP-positive cells than infection with AdGFP. Delta-24-RGD was more cytopathic to both low- and high-CAR-expressing glioma lines than Delta-24, and it replicated more efficiently in both cell lines. In the xenografted mice, intratumoral injection of Delta-24-RGD was associated with longer survival than intratumoral injection of Delta-24 (P<.001, log-rank test). Furthermore, 60% of Delta-24-RGD-treated mice but only 15% of Delta-24-treated mice survived more than 4 months (difference = 45%, 95% CI = 21% to 68%). CONCLUSIONS: The antitumor activity of Delta-24-RGD suggests that it has the potential to be an effective agent in the treatment of gliomas. PMID- 12734317 TI - Multiplex biomarker approach for determining risk of prostate-specific antigen defined recurrence of prostate cancer. AB - BACKGROUND: Molecular signatures in cancer tissue may be useful for diagnosis and are associated with survival. We used results from high-density tissue microarrays (TMAs) to define combinations of candidate biomarkers associated with the rate of prostate cancer progression after radical prostatectomy that could identify patients at high risk for recurrence. METHODS: Fourteen candidate biomarkers for prostate cancer for which antibodies are available included hepsin, pim-1 kinase, E-cadherin (ECAD; cell adhesion molecule), alpha-methylacyl coenzyme A racemase, and EZH2 (enhancer of zeste homolog 2, a transcriptional repressor). TMAs containing more than 2000 tumor samples from 259 patients who underwent radical prostatectomy for localized prostate cancer were studied with these antibodies. Immunohistochemistry results were evaluated in conjunction with clinical parameters associated with prostate cancer progression, including tumor stage, Gleason score, and prostate-specific antigen (PSA) level. Recurrence was defined as a postsurgery PSA level of more than 0.2 ng/mL. All statistical tests were two-sided. RESULTS: Moderate or strong expression of EZH2 coupled with at most moderate expression of ECAD (i.e., a positive EZH2:ECAD status) was the biomarker combination that was most strongly associated with the recurrence of prostate cancer. EZH2:ECAD status was statistically significantly associated with prostate cancer recurrence in a training set of 103 patients (relative risk [RR] = 2.52, 95% confidence interval [CI] = 1.09 to 5.81; P =.021), in a validation set of 80 patients (RR = 3.72, 95% CI = 1.27 to 10.91; P =.009), and in the combined set of 183 patients (RR = 2.96, 95% CI = 1.56 to 5.61; P<.001). EZH2:ECAD status was statistically significantly associated with disease recurrence even after adjusting for clinical parameters, such as tumor stage, Gleason score, and PSA level (hazard ratio = 3.19, 95% CI = 1.50 to 6.77; P =.003). CONCLUSION: EZH2:ECAD status was statistically significantly associated with prostate cancer recurrence after radical prostatectomy and may be useful in defining a cohort of high-risk patients. PMID- 12734318 TI - Association between osteosarcoma and deleterious mutations in the RECQL4 gene in Rothmund-Thomson syndrome. AB - BACKGROUND: Rothmund-Thomson syndrome (RTS) is an autosomal recessive disorder associated with an increased predisposition to osteosarcoma. Children with RTS typically present with a characteristic skin rash (poikiloderma), small stature, and skeletal dysplasias. Mutations in the RECQL4 gene, which encodes a RecQ DNA helicase, have been reported in a few RTS patients. We examined whether a predisposition to developing osteosarcoma among an international cohort of RTS patients was associated with a distinctive pattern of mutations in the RECQL4 gene. METHODS: We obtained clinical information about and biologic samples from 33 RTS patients (age range = 1-30 years). Eleven patients were diagnosed with osteosarcoma. All 21 exons and 13 short introns of the RECQL4 gene were sequenced from the genomic DNA of all subjects. Kaplan-Meier survival analysis was used to estimate the incidence of osteosarcoma among patients with and without mutations predicted to produce a truncated RECQL4 protein. RESULTS: Twenty-three RTS patients, including all 11 osteosarcoma patients, carried at least one of 19 truncating mutations in their RECQL4 genes. The incidence of osteosarcoma was 0.00 per year in truncating mutation-negative patients (100 person-years of observation) and 0.05 per year in truncating mutation-positive patients (230 person-years of observation) (P =.037; two-sided log-rank test). CONCLUSIONS: Mutations predicted to result in the loss of RECQL4 protein function occurred in approximately two-thirds of RTS patients and are associated with risk of osteosarcoma. Molecular diagnosis has the potential to identify those children with RTS who are at high risk of this cancer. PMID- 12734319 TI - 2-[11C]thymidine positron emission tomography as an indicator of thymidylate synthase inhibition in patients treated with AG337. AB - BACKGROUND: Some anticancer drugs inhibit thymidylate synthase (TS), a key enzyme for thymidine nucleotide biosynthesis. Cells can compensate for depleted thymidine levels by taking up extracellular thymidine via a salvage pathway. We investigated the use of 2-[11C]thymidine positron emission tomography (PET) to measure thymidine salvage kinetics in vivo in humans. METHODS: Five patients with advanced gastrointestinal cancer were PET scanned both before and 1 hour after oral administration of the TS inhibitor AG337 (THYMITAQ [nolatrexed]); seven control patients were scanned twice but not treated with AG337. Thymidine salvage kinetics were measured in vivo using 2-[11C]thymidine PET and spectral analysis to obtain the standardized uptake values (SUV), the area under the time-activity curve (AUC), and the fractional retention of thymidine (FRT). Changes in PET parameters between scans in the AG337-treated and control groups were compared using the Mann-Whitney U test. The relationship between AG337 exposure and AG337 induced changes in tumor FRT and in plasma deoxyuridine levels (a conventional pharmacodynamic systemic measure of TS inhibition) was examined using Spearman's regression analysis. Statistical tests were two-sided. RESULTS: The between-scan change in FRT in patients treated with AG337 (38% increase, 95% confidence interval [CI] = 8% to 68%) was higher than that in control patients (3% increase, 95% CI = -11% to 17%) (P =.028). The level of AG337-induced increase in both 2 [11C]thymidine FRT and plasma deoxyuridine levels was statistically significantly correlated with AG337 exposure (r = 1.00, P =.01 for both). CONCLUSIONS: AG337 administration was associated with increased tumor tracer retention that was consistent with tumor cell uptake of exogenous 2-[11C]thymidine as a result of TS inhibition. 2-[11C]Thymidine PET can be used to measure thymidine salvage kinetics directly in the tissue of interest. PMID- 12734320 TI - Re: Trends in use of adjuvant multi-agent chemotherapy and tamoxifen for breast cancer in the United States: 1975-1999. PMID- 12734323 TI - Re: Is less more? Lessons in radiation schedules in breast cancer. PMID- 12734321 TI - Re: Trends in use of adjuvant multi-agent chemotherapy and tamoxifen for breast cancer in the United States: 1975-1999. PMID- 12734326 TI - Re: Trends in U.S. pleural mesothelioma incidence rates following simian virus 40 contamination of early poliovirus vaccines. PMID- 12734325 TI - Re: Trends in U.S. pleural mesothelioma incidence rates following simian virus 40 contamination of early poliovirus vaccines. PMID- 12734329 TI - Cutting edge: the NK cell receptor 2B4 augments antigen-specific T cell cytotoxicity through CD48 ligation on neighboring T cells. AB - 2B4 is expressed on all NK and a subset of memory/effector CD8(+) T cells. 2B4 binds to CD48 and activates NK cytotoxicity, but its function on CD8(+) T cells is not clear. Furthermore, two isoforms of 2B4 (2B4S and 2B4L) exist in mice but the role of individual isoforms is not known. To address these questions, we generated primary T cell cultures from L(d)-specific 2C/Rag2(-/-) TCR transgenic mice and transduced them with 2B4S or 2B4L. 2B4S- or 2B4L-transduced T cells showed greater cytotoxicity over control cells against CD48(+) and CD48(-) targets, suggesting that ligation of 2B4 by CD48 on target cells was not necessary for 2B4 function. Rather, 2B4/CD48 interaction on adjacent T cells appeared to be critical for cytotoxicity. Therefore, 2B4 functions as a costimulator of CD8(+) T cells in MHC-restricted cytotoxicity. We conclude that 2B4/CD48 interactions among T cells themselves can augment CTL lysis of their specific targets. PMID- 12734330 TI - Cutting edge: role of IL-27/WSX-1 signaling for induction of T-bet through activation of STAT1 during initial Th1 commitment. AB - WSX-1 is a member of the class I cytokine receptor family with homology to IL-12R beta 2 and is essential for the initial mounting of Th1 responses. STAT1 interacts with tyrosine-phosphorylated WSX-1, and the conserved tyrosine residue of the cytoplasmic domain of WSX-1 is essential for transcriptional activation of STAT1. IL-27 stimulation induced STAT1 phosphorylation in wild-type but not in WSX-1-deficient naive CD4(+) T cells. Although IL-27 did not directly induce IFN gamma production by wild-type CD4(+) T cells, IL-12-dependent IFN-gamma production was augmented by IL-27 stimulation in wild-type naive CD4(+) T cells but was impaired in WSX-1-deficient naive CD4(+) T cells. Additionally, IL-27 stimulation induced T-bet and IL-12R beta 2 expression in wild-type, but not in WSX-1-deficient, CD4(+) T cells. Thus, during the initiation of Th1 differentiation, the IL-27/WSX-1 signaling system plays a pivotal role by STAT1 mediated T-bet induction before the IL-12R system. PMID- 12734331 TI - Cutting edge: dependence of TCR antagonism on Src homology 2 domain-containing protein tyrosine phosphatase activity. AB - The mechanism by which antagonist peptides inhibit T cell responses is unknown. Mice deficient in Src homology 2 domain-containing protein tyrosine phosphatase (SHP-1) have revealed its importance in the negative regulation of lymphocyte signaling. We investigated a possible role for SHP-1 in T cell antagonism and demonstrate, for the first time, a substantial increase in SHP-1 activity during antagonism of CD4(+) T cells. Furthermore, the removal of functional SHP-1 prevents antagonism in these cells. Our data demonstrate that T cell antagonism occurs via a negative intracellular signal that is mediated by SHP-1. PMID- 12734332 TI - Environmental chemical-induced pro/pre-B cell apoptosis: analysis of c-Myc, p27Kip1, and p21WAF1 reveals a death pathway distinct from clonal deletion. AB - Polycyclic aromatic hydrocarbons (PAH) are common environmental pollutants that suppress the immune system in part by inducing pro/pre-B cell apoptosis. The PAH induced death signaling pathway resembles the signaling cascade activated during clonal deletion and modeled by B cell receptor cross-linking or by dexamethasone exposure of immature surface Ig(+) B cells in that apoptosis is mediated by NF kappa B down-regulation. Because a PAH-induced, clonally nonrestricted deletion of B cells would have important implications for B cell repertoire development, the nature of the PAH-induced intracellular death signal was studied further. Particular emphasis was placed on the roles of growth arrest and c-Myc, p27(Kip1), and p21(WAF1) expression, because all of these elements contribute to clonal deletion. As in clonal deletion models, and as predicted by the down regulation of NF-kappa B, PAH-induced death of pro/pre-B cells was at least partially dependent on c-Myc down-regulation. Furthermore, whereas dexamethasone induced a G(0)/G(1) cell cycle arrest, PAH had no effect on pro/pre-B cell growth, indicating that growth arrest and apoptosis occur by separable signaling pathways in this early phase of B cell development. Finally, in contrast to clonal deletion, PAH-induced pro/pre-B cell death was not dependent on p27(Kip1) or p21(WAF1) up-regulation but did coincide with p53 induction. These results distinguish the PAH-induced apoptosis pathway from that activated during clonal deletion and indicate that signaling cascades leading to growth arrest and/or apoptosis in pro/pre-B cells differ from those active at later B cell developmental stages. PMID- 12734333 TI - Induction of tumor cell apoptosis in vivo increases tumor antigen cross presentation, cross-priming rather than cross-tolerizing host tumor-specific CD8 T cells. AB - Cross-presentation of cell-bound Ags from established, solid tumors to CD8 cells is efficient and likely to have a role in determining host response to tumor. A number of investigators have predicted that when tumor Ags are derived from apoptotic cells either no response, due to Ag "sequestration," or CD8 cross tolerance would ensue. Because the crucial issue of whether this happens in vivo has never been addressed, we induced apoptosis of established hemagglutinin (HA) transfected AB1 tumors in BALB/c mice using the apoptosis-inducing reagent gemcitabine. This shrank the tumor by approximately 80%. This induction of apoptosis increased cross-presentation of HA to CD8 cells yet neither gross deletion nor functional tolerance of HA-specific CD8 cells were observed, based on tetramer analysis, proliferation of specific CD8 T cells, and in vivo CTL activity. Interestingly, apoptosis primed the host for a strong antitumor response to a second, virus-generated HA-specific signal in that administration of an HA-expressing virus after gemcitabine administration markedly decreased tumor growth compared with viral administration without gemcitabine. Thus tumor cell apoptosis in vivo neither sequesters tumor Ags nor cross-tolerizes tumor specific CD8 cells. This observation has fundamental consequences for the development of tumor immunotherapy protocols and for understanding T cell reactivity to tumors and the in vivo immune responses to apoptotic cells. PMID- 12734334 TI - Two regions of down-regulation in the IgE-mediated signaling pathway in human basophils. AB - Previous studies demonstrated that after stimulation of human basophils with a polyclonal anti-IgE Ab, early signaling elements showed sustained phosphorylation, whereas later elements were transient, suggesting that a region of down-regulation involved inhibition of phosphatidylinositol (PI) 3 kinase or its products. However, the current studies show that under some conditions, syk phosphorylation is transient. Generally, stimulation with a variety of Ags makes this early form of down-regulation more apparent. An exploration of the conditions needed to induce early down-regulation indicates that both the nature of aggregation and the cell surface density of IgE play roles. It was also found that the previously described late form of down-regulation (PI3 kinase product transience) can occur in cells displaying early down-regulation (transient syk phosphorylation), but this phenomenon is revealed by testing for subsequent down regulation of the response to non-cross-reacting stimuli, altering their ability to induce phosphorylation of Akt or extracellular signal-regulated kinase. In contrast, phosphorylation of syk kinase, in response to a non-cross-reacting stimulus, was relatively unaffected by prior stimulation. The magnitude of cross desensitization of the Akt or extracellular signal-regulated kinase response was a function of the strength of the first stimulus. Mediator release showed a similar cross-desensitization effect. Therefore, stimulation induces two forms of down-regulation, one operating before or at the level of syk phosphorylation, possibly characterizing the process formerly known as specific desensitization, and one that operates in the region of PI3 kinase, accounting for the process formerly known as nonspecific desensitization, which is dependent on the strength of stimulus. PMID- 12734335 TI - The lymphoid past of mouse plasmacytoid cells and thymic dendritic cells. AB - There has been controversy over the possible lymphoid origin of certain dendritic cell (DC) subtypes. To resolve this issue, DC and plasmacytoid pre-DC isolated from normal mouse tissues were analyzed for transient (mRNA) and permanent (DNA rearrangement) markers of early stages of lymphoid development. About 27% of the DNA of CD8(+) DC from thymus, and 22-35% of the DNA of plasmacytoid pre-DC from spleen and thymus, was found to contain IgH gene D-J rearrangements, compared with 40% for T cells. However, the DC DNA did not contain IgH gene V-D-J rearrangements nor T cell Ag receptor beta gene D-J rearrangements. The same DC lineage populations containing IgH D-J rearrangements expressed mRNA for CD3 chains, and for pre-T alpha. In contrast, little of the DNA of the conventional DC derived from spleen, lymph nodes, or skin, whether CD8(+) or CD8(-), contained IgH D-J rearrangements and splenic conventional DC expressed very little CD3 epsilon or pre-T alpha mRNA. Therefore, many plasmacytoid pre-DC and thymic CD8(+) DC have shared early steps of development with the lymphoid lineages, and differ in origin from conventional peripheral DC. PMID- 12734336 TI - Regulation of CD8+ T cells undergoing primary and secondary responses to infection in the same host. AB - Naive Ag-specific CD8(+) T cells expand, contract, and become memory cells after infection and/or vaccination. Memory CD8(+) T cells provide faster, more effective secondary responses against repeated exposure to the same pathogen. Using an adoptive transfer system with low numbers of trackable nontransgenic memory CD8(+) T cells, we showed that secondary responses can be comprised of both primary (naive) and secondary (memory) CD8(+) T cells after bacterial (Listeria monocytogenes) and/or viral (lymphocytic choriomeningitis virus) infections. The level of memory CD8(+) T cells present at the time of infection inversely correlated with the magnitude of primary CD8(+) T cell responses against the same epitope but directly correlated with the level of protection against infection. However, similar numbers of Ag-specific CD8(+) T cells were found 8 days postinfection no matter how many memory cells were present at the time of infection. Rapid contraction of primary CD8(+) T cell responses was not influenced by the presence of memory CD8(+) T cells. However, contraction of secondary CD8(+) T cell responses was markedly prolonged compared with primary responses in the same host mice. This situation occurred in response to lymphocytic choriomeningitis virus or L. monocytogenes infection and for CD8(+) T cell responses against multiple epitopes. The delayed contraction of secondary CD8(+) T cells was also observed after immunization with peptide-coated dendritic cells. Together, the results show that the level of memory CD8(+) T cells influences protective immunity and activation of naive precursors specific for the same epitope but has little impact on the magnitude or program of the CD8(+) T cell response. PMID- 12734337 TI - Prostaglandin D2 affects the maturation of human monocyte-derived dendritic cells: consequence on the polarization of naive Th cells. AB - Among the factors produced at inflammatory sites and those capable of modulating dendritic cell (DC) functions, PGD(2) may be important in the outcome of immune responses. The biological roles for PGD(2) are in part effected through two plasma membrane G protein-coupled receptors: the D prostanoid (DP) receptor and the chemoattractant receptor-homologous molecule expressed on Th2 lymphocytes (CRTH2). In this report, we studied the effects of PGD(2) and of its major physiological metabolite, 15-deoxy-Delta(12,14)-PGJ(2) (15d-PGJ(2)), on the functions of human monocyte-derived DC. First, we show that PGD(2) exerts in vitro chemotactic effects on monocytes via CRTH2 activation while it inhibits the chemokine-driven migration of monocyte-derived DC through DP. We also report that PGD(2) and 15d-PGJ(2) alter the LPS- and allergen-induced DC maturation and enhance the CD80/CD86 ratio on mature DC in a DP- and CRTH2-independent manner. Moreover, PGD(2) and 15d-PGJ(2) strongly reduce the secretion of the Th1 promoting cytokine IL-12 and affect the synthesis of chemokines involved in Th1 cell chemotaxis, particularly CXCL10. Inhibition of cytokine/chemokine secretion implicates at least in part DP, but not CRTH2. The effects exerted by PGD(2) are associated with the phosphorylation of CREB, but do not parallel with the deactivation of the NF-kappa B and mitogen-activated protein kinase pathways. In contrast, 15d-PGJ(2) seems to target other cellular proteins. Finally, in a model of Th CD45RA(+) differentiation induced by allergen- and superantigen-pulsed DC, PGD(2) impacts on the orientation of the immune response by favoring a Th2 response. PMID- 12734338 TI - Differential roles of CB1 and CB2 cannabinoid receptors in mast cells. AB - Cannabinoid modulation of immune responses is a pathological consequence of marijuana abuse and a potential outcome of therapeutic application of the drug. Moreover, endogenous cannabinoids are physiological immune regulators. In the present report, we describe alterations in gene transcription that occur after cannabinoid exposure in a mast cell line, RBL2H3. Cannabinoid exposure causes marked changes in the transcript levels for numerous genes, acting both independently of and in concert with immunoreceptor stimulation via Fc epsilon RI. In two mast cell lines, we observed mRNA and protein expression corresponding to both CB1 and CB2 cannabinoid receptor isoforms, contrary to the prevailing view that CB1 is restricted to the CNS. We show that coexpression of the two isoforms is not functionally redundant in mast cells. Analysis of signaling pathways downstream of cannabinoid application reveals that activation of extracellular signal-regulated kinase, AKT, and a selected subset of AKT targets is accomplished by CB2 ligands and nonselective CB1/CB2 agonists in mast cells. CB1 inhibition does not affect AKT or extracellular signal-regulated kinase activation by cannabinoids, indicating that CB2 is the predominant regulatory receptor for these kinases in this cell context. CB1 receptors are, however, functional in these mast cells, since they can contribute to suppression of secretory responses. PMID- 12734339 TI - Stochastic model of T cell proliferation: a calculus revealing IL-2 regulation of precursor frequencies, cell cycle time, and survival. AB - The outcome of Ag exposure is dictated by complex regulation of T cell proliferation. The rates of proliferation and survival are altered by numerous signals that the cell receives and integrates to achieve a net response. We have illustrated previously how small changes in kinetic parameters can lead to large differences, even under conditions of saturating IL-2. In this study, we examine the effect of varying IL-2 concentration on T cell response and develop a model incorporating additional parameters of proliferation and survival. Strikingly, the proportion of cells that enter the first division, but not the time at which they enter, is dramatically altered by IL-2. Furthermore, the survival and average division time of cells in later divisions are also altered by IL-2 concentration. Together, the small simultaneous effects on these parameters result in large differences in total cell number. These results reveal how in vitro systems may exaggerate the contribution of IL-2, and thus how costimuli or additional helper cells that alter IL-2 concentration, even by relatively small amounts, will generate large in vitro differences in cell number and therefore appear obligatory. Furthermore, they illustrate how a quantitative model of T cell activation can clarify how complex signal integration is handled by T cells in situ, and therefore more appropriately aid development of a theory of behavior. PMID- 12734341 TI - Histone deacetylation, but not hypermethylation, modifies class II transactivator and MHC class II gene expression in squamous cell carcinomas. AB - In this study, we first categorized nine squamous cell carcinoma (SCC) cell lines into two groups in terms of the expression of HLA-DR, -DP, and -DQ molecules. Subsequently, the expression of class II transactivator (CIITA) was studied in these cell lines, because it is widely accepted that the expression of MHC class II molecules is regulated by different types of CIITA transcripts that are initiated by distinct promoters. The majority of the SCC cell lines (six of nine) expressed HLA-DR molecules and CIITA promoter IV (pIV) transcripts in the presence of IFN-gamma. In contrast, three of the nine SCC cell lines were completely negative for class II molecules and all types of CIITA, suggesting epigenetic changes in the promoter region in these cells. Previously, methylation of CIITA pIV was reported to silence CIITA gene expression. We extensively studied the methylation status of CIITA pIV using a panel of 22 SCC cell lines. Remarkably, none of the SCC cell lines demonstrated hypermethylation at the site. In contrast, treatment with a histone deacetylation inhibitor in combination with IFN-gamma clearly restored the expression of the CIITA type IV gene in the HLA-DR negative SCC cell lines, and the acetylation status of histone H3 examined by chromatin immunoprecipitation analysis was closely associated with the gene expression. Moreover, stable transfection of the CIITA gene into an HLA-DR negative cell line restored constitutive expression of MHC class II molecules. Therefore, histone deacetylation, but not hypermethylation, modifies CIITA DNA and class II gene expression in SCC. PMID- 12734340 TI - Active form of Notch members can enforce T lymphopoiesis on lymphoid progenitors in the monolayer culture specific for B cell development. AB - The in vitro induction of T lymphopoiesis needs the precise stereoscopic structure of thymus tissues as seen in fetal thymus organ culture. In this study, we demonstrated for the first time that the introduction of the intracellular region of Notch1 can induce T cells expressing TCR without any thymic environment. In the coculture on the monolayer of OP-9, which was originally known to support B cell specific development, hemopoietic progenitors developed into Thy-1(+)CD25(+) T lineage cells if the progenitor cells were infected with the retrovirus containing Notch1 intracellular domains. The Thy-1(+) cells progressed to a further developmental stage, CD4 and CD8 double-positive cells expressing TCR on the cell surface, if they were further cultured on OP-9 or in the thymus. However, T cell induction by intracellular Notch1 failed unless both OP-9 and IL-7 were present. It is notable that Notch2 and Notch3 showed an effect on T lymphopoiesis similar to that of Notch1. These results indicate that in vitro T lymphopoiesis is inducible by signaling via Notch family members in a lineage-specific manner but shares other stroma-derived factors including IL-7 with B lymphopoiesis. PMID- 12734342 TI - Differential control of cell cycle, proliferation, and survival of primary T lymphocytes by purine and pyrimidine nucleotides. AB - Purine and pyrimidine nucleotides play critical roles in DNA and RNA synthesis as well as in membrane lipid biosynthesis and protein glycosylation. They are necessary for the development and survival of mature T lymphocytes. Activation of T lymphocytes is associated with an increase of purine and pyrimidine pools. However, the question of how purine vs pyrimidine nucleotides regulate proliferation, cell cycle, and survival of primary T lymphocytes following activation has not yet been specifically addressed. This was investigated in the present study by using well-known purine (mycophenolic acid, 6-mercaptopurine) and pyrimidine (methotrexate, 5-fluorouracil) inhibitors, which are used in neoplastic diseases or as immunosuppressive agents. The effect of these inhibitors was analyzed according to their time of addition with respect to the initiation of mitogenic activation. We showed that synthesis of both purine and pyrimidine nucleotides is required for T cell proliferation. However, purine and pyrimidine nucleotides differentially regulate the cell cycle since purines control both G(1) to S phase transition and progression through the S phase, whereas pyrimidines only control progression from early to intermediate S phase. Furthermore, inhibition of pyrimidine synthesis induces apoptosis whatever the time of inhibitor addition whereas inhibition of purine nucleotides induces apoptosis only when applied to already cycling T cells, suggesting that both purine and pyrimidine nucleotides are required for survival of cells committed into S phase. These findings reveal a hitherto unknown role of purine and pyrimidine de novo synthesis in regulating cell cycle progression and maintaining survival of activated T lymphocytes. PMID- 12734343 TI - Mechanisms of accelerated immune-mediated diabetes resulting from islet beta cell expression of a Fas ligand transgene. AB - Nonobese diabetic (NOD) mice transgenic for Fas ligand (FasL) on islet beta cells (HIPFasL mice) exhibit an accelerated diabetes distinct from the normal autoimmune diabetes of NOD mice. This study was undertaken to define the mechanism underlying accelerated diabetes development in HIPFasL mice. It was found that diabetes in HIPFasL mice is dependent on the NOD genetic background, as HIPFasL does not cause diabetes when crossed into other mice strains and is lymphocyte dependent, as it does not develop in HIPFasL(SCID) mice. Diabetes development in NOD(SCID) recipients of diabetic HIPFasL splenocytes is slower than when using splenocytes from diabetic NOD mice. Beta cells from HIPFasL mice are more susceptible to cytokine-induced apoptosis than wild-type NOD beta cells, and this can be blocked with anti-FasL Ab. HIPFasL islets are more rapidly destroyed than wild-type islets when transplanted into nondiabetic NOD mice. This confirms that FasL(+) islets do not obtain immune privilege, and instead NOD beta cells constitutively expressing FasL are more susceptible to apoptosis induced by Fas-FasL interaction. These findings are consistent with the accelerated diabetes of young HIPFasL mice being a different disease process from the autoimmune diabetes of wild-type NOD mice. The data support a mechanism by which cytokines produced by the insulitis lesion mediate up-regulation of beta cell Fas expression, resulting in suicide or fratricide of HIPFasL beta cells that overexpress FasL. PMID- 12734344 TI - IL-18 and IL-12 signal through the NF-kappa B pathway to induce NK-1R expression on T cells. AB - Substance P engages the T cell neurokinin 1 receptor (NK-1R) to enhance IFN-gamma production. NK-1R on T cells is inducible. We studied mechanisms regulating T cell NK-1R expression. Murine splenocytes were cultured for 4 h with or without rIL-12 or rIL-18. Both IL-12 and IL-18 induced splenic T cells to express NK-1R transcripts. Induction was blocked by actinomycin D, but not cycloheximide, suggesting that protein synthesis was not required for initiation of NK-1R gene transcription. Inhibition of T cell NF-kappa B activation or NF-kappa B nuclear translocation also blocked NK-1R transcription. IL-12 and IL-18 strongly induce NK-1R mRNA expression in splenocytes from Stat4(-/-) mice, suggesting that the Stat4 pathway was not required for the induction of NK-1R transcription. Splenic T cells exposed to IL-12 or IL-18 in the presence of IL-10 expressed no NK-1R mRNA. However, TGF beta did not prevent NK-1R mRNA expression. Thus, IL-12 and IL 18 induce T cells to express NK-1R through NF-kappa B activation. IL-10, a regulator of the Th1 response, blocks this activation. These data further suggest that SP and NK-1R, which promote IFN-gamma synthesis, are part of the Th1 pathway of immunity. PMID- 12734345 TI - Superantigen-induced regulatory T cells display different suppressive functions in the presence or absence of natural CD4+CD25+ regulatory T cells in vivo. AB - Repeated exposures to both microbial and innocuous Ags in vivo have been reported to both eliminate and tolerize T cells after their initial activation and expansion. The remaining tolerant T cells have been shown to suppress the response of naive T cells in vitro. This feature is reminiscent of natural CD4(+)CD25(+) regulatory T cells. However, it is not known whether the regulatory function of in vivo-tolerized T cells is similar to the function of natural CD4(+)CD25(+) regulatory T cells. In this study, we demonstrate that CD4(+)CD25(+) as well as CD4(+)CD25(-) T cells isolated from mice treated with superantigen three consecutive times to induce tolerance were functionally comparable to natural CD4(+)CD25(+) regulatory T cells, albeit more potent. The different subpopulations of in vivo-tolerized CD4(+) T cells efficiently down modulated costimulatory molecules on dendritic cells, and their suppressive functions were strictly cell contact dependent. Importantly, we demonstrate that conventional CD4(+)CD25(-) T cells could also be induced to acquire regulatory functions by the same regimen in the absence of natural regulatory T cells in vivo, but that such regulatory cells were functionally different. PMID- 12734346 TI - IL-15 promotes the survival of naive and memory phenotype CD8+ T cells. AB - IL-15 stimulates the proliferation of memory phenotype CD44(high)CD8(+) T cells and is thought to play a key role in regulating the turnover of these cells in vivo. We have investigated whether IL-15 also has the capacity to affect the life span of naive phenotype (CD44(low)) CD8(+) T cells. We report that IL-15 promotes the survival of both CD44(low) and CD44(high) CD8(+) T cells, doing so at much lower concentrations than required to induce proliferation of CD44(high) cells. Rescue from apoptosis was associated with the up-regulation of Bcl-2 in both cell types, whereas elevated expression of Bcl-x(L) was observed among CD44(high) but not CD44(low) CD8(+) cells. An investigation into the role of IL-15R subunits in mediating the effects of IL-15 revealed distinct contributions of the alpha- and beta- and gamma-chains. Most strikingly, IL-15R alpha was not essential for either induction of proliferation or promotion of survival by IL-15, but did greatly enhance the sensitivity of cells to low concentrations of IL-15. By contrast, the beta- and gamma-chains of the IL-15R were absolutely required for the proliferative and pro-survival effects of IL-15, although it was not necessary for CD44(high)CD8(+) cells to express higher levels of IL-15R beta than CD44(low) cells to proliferate in response to IL-15. These results show that IL 15 has multiple effects on CD8 T cells and possesses the potential to regulate the life span of naive as well as memory CD8(+) T cells. PMID- 12734347 TI - A peptide from heat shock protein 60 is the dominant peptide bound to Qa-1 in the absence of the MHC class Ia leader sequence peptide Qdm. AB - The MHC class Ib molecule Qa-1 binds specifically and predominantly to a single 9 aa peptide (AMAPRTLLL) derived from the leader sequence of many MHC class Ia proteins. This peptide is referred to as Qdm. In this study, we report the isolation and sequencing of a heat shock protein 60-derived peptide (GMKFDRGYI) from Qa-1. This peptide is the dominant peptide bound to Qa-1 in the absence of Qdm. A Qa-1-restricted CTL clone recognizes this heat shock protein 60 peptide, further verifying that it binds to Qa-1 and a peptide from the homologous Salmonella typhimurium protein GroEL (GMQFDRGYL). These observations have implications for how Qa-1 can influence NK cell and T cell effector function via the TCR and CD94/NKG2 family members, and how this effect can change under conditions that cause the peptides bound to Qa-1 to change. PMID- 12734348 TI - Extracellular matrix conditions T cells for adhesion to tissue interstitium. AB - The activation and differentiation of peripheral blood T cells (PBT) are known to correlate with increased surface expression and adhesive capacity of beta(1) integrins, which mediate adhesion to the extracellular matrix (ECM). However, little is known about the regulation of integrin expression, affinity, and avidity on tissue T cells after they are embedded in the interstitial ECM. In this study we show that tissue T cells, freshly isolated from their residence in the interstitial ECM of the intestinal lamina propria, express a distinct subset of functionally active integrins that contribute to enhanced adhesion to purified collagen, fibronectin, and cell-derived ECM when compared with freshly isolated, short term activated, and long term cultured PBT. Furthermore, integrin usage is distinct between circulating and tissue-derived T cells, in that lamina propria T cells prefer to bind to collagen, while PBT lymphoblasts choose fibronectin when presented with a complex, three-dimensional, cell-derived matrix. To identify the extrinsic factors that regulate the conversion from a nonadhesive PBT to highly adhesive tissue T cell, we demonstrate that activation of PBT in the presence of fibronectin or collagen rapidly generates a surface integrin expression profile, an integrin usage pattern, and adhesive capacity mirroring that of a tissue T cell. These results indicate that the tissue ECM microenvironment instructs newly arrived T cells for further interactions with the underlying matrix and thereby imprints them with a signature tissue adhesive phenotype. PMID- 12734349 TI - Mast cells express novel functional IL-15 receptor alpha isoforms. AB - Mast cells previously have been reported to be regulated by IL-15 and to express a distinct IL-15R, termed IL-15RX. To further examine IL-15 binding and signaling in mast cells, we have studied the nature of the IL-15R and some of its biological activities in these cells. In this study, we report the existence of three novel isoforms of the IL-15R alpha chain in murine bone marrow-derived mast cells as a result of an alternative exon-splicing mechanism within the IL-15R alpha gene. These correspond to new mRNA transcripts lacking exon 4; exons 3 and 4; or exons 3, 4, and 5 (IL-15R alpha Delta 4, IL-15R alpha Delta 3,4, IL-15R alpha Delta 3,4,5). After transient transfection in COS-7 cells, all IL-15R alpha isoforms associate with the Golgi apparatus, the endoplasmic reticulum, the perinuclear space, and the cell membrane. Analysis of glycosylation pattern demonstrates the usage of a single N-glycosylation site, while no O-glycosylation is observed. Importantly, IL-15 binds with high affinity to, and promotes the survival of, murine BA/F3 cells stably transfected with the IL-15R alpha isoforms. Furthermore, we report that signaling mediated by IL-15 binding to the newly identified IL-15R alpha isoforms involves the phosphorylation of STAT3, STAT5, STAT6, Janus kinase 2, and Syk kinase. Taken together, our data indicate that murine mast cells express novel, fully functional IL-15R alpha isoforms, which can explain the selective regulatory effects of IL-15 on these cells. PMID- 12734350 TI - Attenuation of immunological symptoms of allergic asthma in mice lacking the tyrosine kinase ITK. AB - Allergic asthma patients manifest airway inflammation and some show increases in eosinophils, T(H)2 cells, and cytokines, increased mucous production in the lung, and elevated serum IgE. This T(H)2-type response suggests a prominent role for T(H)2 cells and their cytokines in the pathology of this disease. The Tec family nonreceptor tyrosine kinase inducible T cell kinase (ITK) has been shown to play a role in the differentiation and/or function of T(H)2-type cells, suggesting that ITK may represent a good target for the control of asthma. Using a murine model of allergic asthma, we show here that ITK is involved in the development of immunological symptoms seen in this model. We show that mice lacking ITK have drastically reduced lung inflammation, eosinophil infiltration, and mucous production following induction of allergic asthma. Notably, T cell influx into the lung was reduced in mice lacking ITK. T cells from ITK(-/-) mice also exhibited reduced proliferation and cytokine secretion, in particular IL-5 and IL 13, in response to challenge with the allergen OVA, despite elevated levels of total IgE and increased OVA-specific IgE responses. Our results suggest that the tyrosine kinase ITK preferentially regulates the secretion of the T(H)2 cytokines IL-5 and IL-13 and may be an attractive target for antiasthmatic drugs. PMID- 12734351 TI - Nitric oxide-independent CTL suppression during tumor progression: association with arginase-producing (M2) myeloid cells. AB - Most of the mice bearing a s.c. BW-Sp3 lymphoma tumor mount a CD8(+) T cell mediated response resulting in tumor regression. Nonetheless, tumor progression occurs in some of the recipients and is associated with CTL inactivity. We demonstrated that T cell-activating APC were induced in regressors whereas T cell suppressive myeloid cells predominated in the spleen of progressors. Indeed, in vitro depletion of either the adherent or the CD11b(+) populations restored T cell cytotoxicity and proliferation in these mice. This CTL inhibition was cell to-cell contact-dependent but not mediated by NO. However, the same progressor suppressive cells prevented the activity of in vitro-restimulated CTLs derived from regressors in a cell-to-cell contact and NO-dependent fashion. Thus, either the NO-dependent or -independent suppressive pathway prevailed, depending on the target CTL population. In addition, the suppressive population expressed a high arginase activity, suggesting an association of the suppressive phenotype with alternatively activated (M2) myeloid cells. However, the high arginase activity is not directly involved in the suppressive process. Our results provide new insights for myeloid cell-mediated CTL inhibition during cancer progression. PMID- 12734352 TI - Early IL-2 production by mouse dendritic cells is the result of microbial-induced priming. AB - Dendritic cells (DCs) are professional APCs able to initiate innate and adaptive immune responses against invading pathogens. Different properties such as the efficient Ag processing machinery, the high levels of expression of costimulatory molecules and peptide-MHC complexes, and the production of cytokines contribute in making DCs potent stimulators of naive T cell responses. Recently we have observed that DCs are able to produce IL-2 following bacterial stimulation, and we have demonstrated that this particular cytokine is a key molecule conferring to early bacterial activated DCs unique T cell priming capacity. In the present study we show that many different microbial stimuli, but not inflammatory cytokines, are able to stimulate DCs to produce IL-2, indicating that DCs can distinguish a cytokine-mediated inflammatory process from the actual presence of an infection. The capacity to produce IL-2 following a microbial stimuli encounter is a feature shared by diverse DC subtypes in vivo, such as CD8 alpha(+) and CD8 alpha(-) splenic DCs and epidermal Langerhans cells. When early activated DCs interact with T cells, IL-2 produced by DCs is enriched at the site of cell-cell contact, confirming the importance of DCs-derived IL-2 in T cell activation. PMID- 12734354 TI - A subset of human dendritic cells in the T cell area of mucosa-associated lymphoid tissue with a high potential to produce TNF-alpha. AB - Recently, a new class of human dendritic cell (DC) precursors has been described in the peripheral blood recognized by the mAb M-DC8. These cells represent approximately 1% of PBMC and acquire several characteristics of myeloid DC upon in vitro culture. In this report we show that M-DC8(+) monocytes secrete in response to LPS >10 times the amount of TNF-alpha as M-DC8(-) monocytes, but produce significantly less IL-10. Consistent with a role in inflammatory responses, we found that M-DC8(+) cells localized in the T cell area of inflamed human tonsils and in the subepithelial dome region of Peyer's patches. In patients with active Crohn's disease, abundant M-DC8(+) cells were detectable in inflamed ileal mucosa, which were entirely depleted after systemic steroid treatment. Our results indicate that M-DC8(+) cells are cells of DC phenotype in inflamed mucosa-associated lymphoid tissue that may contribute to the high level of TNF-alpha production in Crohn's disease. We infer that selective elimination of M-DC8(+) cells in inflammatory diseases has therapeutic potential. PMID- 12734353 TI - TNF enhances CD4+ T cell alloproliferation, IFN-gamma responses, and intestinal graft-versus-host disease by IL-12-independent mechanisms. AB - Inhibition of TNF/TNFR2 interactions ameliorates intestinal graft-vs-host disease (GVHD) and Th1 cytokine responses induced by transfer of B6 CD4(+) spleen cells into irradiated MHC class II disparate B6.C-H-2(bm12) (bm12) x B6 F(1) recipients. The present studies examined whether these effects of TNF are IL-12 dependent. T cell proliferative responses of B6.129S1-IL-12rb2(tm1Jm) (B6.IL-12R( /-)) responder spleen cells were found to be comparable to those of control B6 spleen cells. TNF inhibition reduced T cell proliferation and IFN-gamma production in supernatants of MLC using either B6.IL-12R(-/-) or control B6 responder cells. GVHD induced wasting disease in recipients of B6.IL-12R(-/-) CD4(+) spleen cells that received a TNF inhibitor-encoding adenovirus (5.4 +/- 6.5% weight loss (n = 7)) was significantly reduced compared with levels of weight loss observed in recipients that had received a control adenovirus (25.7 +/- 12.2% weight loss (n = 11), p = 0.001). Furthermore, TNF inhibition was associated with a reduction in colonic GVHD scores (p = 0.039) and in the percentage of the splenic CD4(+) T cells that expressed IFN-gamma (16 vs 6%). These findings indicate that TNF promotes CD4(+) T cell alloproliferation, IFN gamma responses, and intestinal GVHD by IL-12-independent mechanisms. PMID- 12734355 TI - Repertoire shift in the humoral response to phosphocholine-keyhole limpet hemocyanin: VH somatic mutation in germinal center B cells impairs T15 Ig function. AB - Phosphocholine (PC) is a naturally occurring Ag common to many pathogenic microorganisms. Early in the primary response to PC conjugated to keyhole limpet hemocyanin (KLH), T15 Id(+) Abs constitute >90% of the serum Ig in BALB/c mice. During the late primary and memory response to PC-protein, a shift in the repertoire occurs and T15 Id(+) Abs lose dominance. In this study, we use immunohistochemistry and single germinal center microdissection to locate T15 Id(+) cells in the spleen in a primary response to PC-KLH. We demonstrate T15 Id(+) B cells and V(H)1-DFL16.1-JH1 and V kappa 22-J kappa 5 rearrangements in germinal centers early in the immune response; thus loss of T15 dominance is not due to lack of T15 cells within germinal centers. One-hundred thirty one V(H)1 and 57 V kappa 22 rearrangements were cloned and sequenced. Thirty four percent of the V(H)1 clones and 37% of the V kappa 22 clones contained somatic mutations indicating participation in the germinal center response. Six variant T15 H clones were expressed with wild-type T15 L chain in vitro. Two of these Abs were defective in secretion providing the first evidence that mutation occurring in vivo can disrupt Ig assembly and secretion. Of the four secretion-competent Abs, two failed to display binding to PC-protein, while the other two displayed altered carrier recognition. These results indicate that somatic mutation of T15 in vivo can result in the loss of binding and secretion, potentially leading to B cell wastage. The failure of T15 to gain affinity enhancing mutations in the face of these detrimental changes may contribute to repertoire shift. PMID- 12734356 TI - Prevalent role of TCR alpha-chain in the selection of the preimmune repertoire specific for a human tumor-associated self-antigen. AB - The specificity of recognition of pMHC complexes by T lymphocytes is determined by the V regions of the TCR alpha- and beta-chains. Recent experimental evidence has suggested that Ag-specific TCR repertoires may exhibit a more V alpha- than V beta-restricted usage. Whether V alpha usage is narrowed during immune responses to Ag or if, on the contrary, restricted V alpha usage is already defined at the early stages of TCR repertoire selection, however, has remained unexplored. Here, we analyzed V and CDR3 TCR regions of single circulating naive T cells specifically detected ex vivo and isolated with HLA-A2/melan-A peptide multimers. Similarly to what was previously observed for melan-A-specific Ag-experienced T cells, we found a relatively wide V beta usage, but a preferential V alpha 2.1 usage. Restricted V alpha 2.1 usage was also found among single CD8(+) A2/melan-A multimer(+) thymocytes, indicating that V alpha-restricted selection takes place in the thymus. V alpha 2.1 usage, however, was independent from functional avidity of Ag recognition. Thus, interaction of the pMHC complex with selected V alpha-chains contributes to set the broad Ag specificity, as underlined by preferential binding of A2/melan-A multimers to V alpha 2.1-bearing TCRs, whereas functional outcomes result from the sum of these with other interactions between pMHC complex and TCR. PMID- 12734357 TI - Decreased binding of peptides-MHC class I (pMHC) multimeric complexes to CD8 affects their binding avidity for the TCR but does not significantly impact on pMHC/TCR dissociation rate. AB - The CD8 coreceptor plays a crucial role in both T cell development in the thymus and in the activation of mature T cells in response to Ag-specific stimulation. In this study we used soluble peptides-MHC class I (pMHC) multimeric complexes bearing mutations in the CD8 binding site that impair their binding to the MHC, together with altered peptide ligands, to assess the impact of CD8 on pMHC binding to the TCR. Our data support a model in which CD8 promotes the binding of TCR to pMHC. However, once the pMHC/TCR complex is formed, the TCR dominates the pMHC/TCR dissociation rates. As a consequence of these molecular interactions, under physiologic conditions CD8 plays a key role in complex formation, resulting in the enhancement of CD8 T cell functions whose specificity, however, is determined by the TCR. PMID- 12734358 TI - Molecular identification and functional expression of mu 3, a novel alternatively spliced variant of the human mu opiate receptor gene. AB - Studies from our laboratory have revealed a novel mu opiate receptor, mu 3, which is expressed in both vascular tissues and leukocytes. The mu 3 receptor is selective for opiate alkaloids and is insensitive to opioid peptides. We now identify the mu 3 receptor at the molecular level using a 441-bp conserved region of the mu 1 receptor. Sequence analysis of the isolated cDNA suggests that it is a novel, alternatively spliced variant of the mu opiate receptor gene. To determine whether protein expressed from this cDNA exhibits the biochemical characteristics expected of the mu 3 receptor, the cDNA clone was expressed in a heterologous system. At the functional level, COS-1 cells transfected with the mu 3 receptor cDNA exhibited dose-dependent release of NO following treatment with morphine, but not opioid peptides (i.e., Met-enkephalin). Naloxone was able to block the effect of morphine on COS-1 transfected cells. Nontransfected COS-1 cells did not produce NO in the presence of morphine or the opioid peptides at similar concentrations. Receptor binding analysis with [(3)H]dihydromorphine further supports the opiate alkaloid selectivity and opioid peptide insensitivity of this receptor. These data suggest that this new mu opiate receptor cDNA encodes the mu 3 opiate receptor, since it exhibits biochemical characteristics known to be unique to this receptor (opiate alkaloid selective and opioid peptide insensitive). Furthermore, using Northern blot, RT-PCR, and sequence analysis, we have demonstrated the expression of this new mu variant in human vascular tissue, mononuclear cells, polymorphonuclear cells, and human neuroblastoma cells. PMID- 12734359 TI - DNA methylation and chromatin structure regulate T cell perforin gene expression. AB - Perforin is a cytotoxic effector molecule expressed in NK cells and a subset of T cells. The mechanisms regulating its expression are incompletely understood. We observed that DNA methylation inhibition could increase perforin expression in T cells, so we examined the methylation pattern and chromatin structure of the human perforin promoter and upstream enhancer in primary CD4(+) and CD8(+) T cells as well as in an NK cell line that expresses perforin, compared with fibroblasts, which do not express perforin. The entire region was nearly completely unmethylated in the NK cell line and largely methylated in fibroblasts. In contrast, only the core promoter was constitutively unmethylated in primary CD4(+) and CD8(+) cells, and expression was associated with hypomethylation of an area residing between the upstream enhancer at -1 kb and the distal promoter at -0.3 kb. Treating T cells with the DNA methyltransferase inhibitor 5-azacytidine selectively demethylated this area and increased perforin expression. Selective methylation of this region suppressed promoter function in transfection assays. Finally, perforin expression and hypomethylation were associated with localized sensitivity of the 5' flank to DNase I digestion, indicating an accessible configuration. These results indicate that DNA methylation and chromatin structure participate in the regulation of perforin expression in T cells. PMID- 12734360 TI - The H4b minor histocompatibility antigen is caused by a combination of genetically determined and posttranslational modifications. AB - Minor histocompatibility (H) Ag disparities result in graft-vs-host disease and chronic solid allograft rejection in MHC-identical donor-recipient combinations. Minor H Ags are self protein-derived peptides presented by MHC class I molecules. Most arise as a consequence of allelic variation in the bound peptide (p) that results in TCR recognizing the p/MHC as foreign. We used a combinational peptide screening approach to identify the immune dominant H2K(b)-restricted epitope defining the mouse H4(b) minor H Ag. H4(b) is a consequence of a P3 threonine to isoleucine change in the MHC-bound peptide derived from epithelial membrane protein-3. This allelic variation also leads to phosphorylation of the H4(b) but not the H4(a) epitope. Further, ex vivo CD8(+) T lymphocytes bind phosphorylated Ag tetramers with high efficiency. Although we document the above process in the minor H Ag system, posttranslational modifications made possible by subtle amino acid changes could also contribute to immunogenicity and immune dominance in tumor immunotherapeutic settings. PMID- 12734361 TI - Selective regulation of mature IgG1 transcription by CD86 and beta 2-adrenergic receptor stimulation. AB - Stimulation of CD86 and the beta(2)-adrenergic receptor (beta(2)AR) on a B cell, either alone or together, is known to increase the level of IgG1 protein produced by a CD40 ligand/IL-4-activated B cell. It is also known that the mechanism by which CD40 and IL-4R stimulation on a B cell increases the level of IgG1 protein is by increasing germline gamma 1 transcription, IgG1 class switching, and mature IgG1 transcription, while the molecular mechanism responsible for mediating the CD86- and beta(2)AR-induced effect remains unknown. In the present study using real-time PCR we show that the level of mature IgG1 transcription increases in CD40 ligand/IL-4-activated B cells following stimulation of either CD86 and/or beta(2)AR, and that this increase reflects the increase in IgG1 protein. Furthermore, we show that the CD86- and/or beta(2)AR-induced increase in mature IgG1 transcript is due to an increase in the rate of mature IgG1 transcription, as determined by nuclear run-on analysis. This effect is additive when both receptors are stimulated and is lost when B cells from CD86- and beta(2)AR deficient mice are used. In contrast, the level of germline gamma 1 transcription, the stability of mature IgG1 transcript, the number of IgG1 positive B cells, and the number of IgG1-secreting B cells did not change. These results provide the first evidence that CD86 and/or beta(2)AR stimulation on a CD40 ligand/IL-4-activated B cell increases the level of IgG1 protein produced per cell by increasing the rate of mature IgG1 transcription. PMID- 12734362 TI - The relative contribution of IL-4 receptor signaling and IL-10 to susceptibility to Leishmania major. AB - The roles of IL-10 and IL-4 receptor signaling were evaluated in a murine model of Leishmania major infection. In previous studies the L. major substrain LV39 caused progressive, nonhealing lesions in BALB/c mice deficient for IL-4R alpha chain (IL-4R alpha), while substrain IR173 was highly controlled. To explore whether IL-10 is responsible for inducing susceptibility to LV39, wild-type and IL-4R alpha(-/-) mice were treated with anti-IL-10R mAb, and in a genetic approach, the IL-4R alpha(-/-) mice were crossed with BALB/c IL-10(-/-) mice. In contrast to the lack of resistance conferred by IL-4R alpha gene deletion, partial resistance to LV39 was conferred by IL-10 gene deletion or treatment of BALB/c mice with anti-IL-10R mAb. Lesion sizes and LV39 parasite numbers were further and dramatically reduced in both anti-IL-10R-treated IL-4R alpha(-/-) mice and IL-4R alpha x IL-10 double knockouts. Anti-IL-10R mAb treatment further suppressed parasite growth in IL-4R alpha(-/-) mice infected with L. major IR173. Production of IFN-gamma was only increased relative to wild-type or littermate controls in IL-4R alpha(-/-) mice with complementary defects in IL-10. Comparisons of IFN-gamma-treated infected macrophages in vitro indicated that LV39 required 25- to 500-fold greater concentrations of IFN-gamma than IR173 infected macrophages to achieve a similar efficiency of parasite killing. These studies suggest that regardless of parasite substrain, IL-10 is as important as IL-4/IL-13 in promoting susceptibility to L. major and even more so for those substrains that are relatively resistant to IFN-gamma mediated killing. PMID- 12734363 TI - Toll-like receptor 2 (TLR2) and TLR9 signaling results in HIV-long terminal repeat trans-activation and HIV replication in HIV-1 transgenic mouse spleen cells: implications of simultaneous activation of TLRs on HIV replication. AB - Opportunistic infections are common in HIV-infected patients; they activate HIV replication and contribute to disease progression. In the present study we examined the role of Toll-like receptor 2 (TLR2) and TLR9 in HIV-long terminal repeat (HIV-LTR) trans-activation and assessed whether TLR4 synergized with TLR2 or TLR9 to induce HIV replication. Soluble Mycobacterium tuberculosis factor (STF) and phenol-soluble modulin from Staphylococcus epidermidis induced HIV-LTR trans-activation in human microvessel endothelial cells cotransfected with TLR2 cDNA. Stimulation of ex vivo spleen cells from HIV-1 transgenic mice with TLR4, TLR2, and TLR9 ligands (LPS, STF, and CpG DNA, respectively) induced p24 Ag production in a dose-dependent manner. Costimulation of HIV-1 transgenic mice spleen cells with LPS and STF or CpG DNA induced TNF-alpha and IFN-gamma production in a synergistic manner and p24 production in an additive fashion. In the THP-1 human monocytic cell line stably expressing the HIV-LTR-luciferase construct, LPS and STF also induced HIV-LTR trans-activation in an additive manner. This is the first time that TLR2 and TLR9 and costimulation of TLRs have been shown to induce HIV replication. Together these results underscore the importance of TLRs in bacterial Ag- and CpG DNA-induced HIV-LTR trans-activation and HIV replication. These observations may be important in understanding the role of the innate immune system and the molecular mechanisms involved in the increased HIV replication and HIV disease progression associated with multiple opportunistic infections. PMID- 12734364 TI - The Toll-like receptor 5 stimulus bacterial flagellin induces maturation and chemokine production in human dendritic cells. AB - Toll-like receptors (TLRs) are pattern recognition receptors that serve an important function in detecting pathogens and initiating inflammatory responses. Upon encounter with foreign Ag, dendritic cells (DCs) go through a maturation process characterized by an increase in surface expression of MHC class II and costimulatory molecules, which leads to initiation of an effective immune response in naive T cells. The innate immune response to bacterial flagellin is mediated by TLR5, which is expressed on human DCs. Therefore, we sought to investigate whether flagellin could induce DC maturation. Immature DCs were cultured in the absence or presence of flagellin and monitored for expression of cell surface maturation markers. Stimulation with flagellin induced increased surface expression of CD83, CD80, CD86, MHC class II, and the lymph node-homing chemokine receptor CCR7. Flagellin stimulated the expression of chemokines active on neutrophils (IL-8/CXC chemokine ligand (CXCL)8, GRO-alpha/CXCL1, GRO beta/CXCL2, GRO-gamma/CXCL3), monocytes (monocyte chemoattractant protein-1/CC chemokine ligand (CCL)2), and immature DCs (macrophage-inflammatory protein-1 alpha/CCL3, macrophage-inflammatory protein-1 beta/CCL4), but not chemokines active on effector T cells (IFN-inducible protein-10 kDa/CXCL10, monokine induced by IFN-gamma/CXCL9, IFN-inducible T cell alpha chemoattractant/CXCL11). However, stimulating DCs with both flagellin and IFN-inducible protein-10 kDa, monokine induced by IFN-gamma, and IFN-inducible T cell alpha chemoattractant expression, whereas stimulation with IFN-beta or flagellin alone failed to induce these chemokines. In functional assays, flagellin-matured DCs displayed enhanced T cell stimulatory activity with a concomitant decrease in endocytic activity. Finally, DCs isolated from mouse spleens or bone marrows were shown to not express TLR5 and were not responsive to flagellin stimulation. These results demonstrate that flagellin can directly stimulate human but not murine DC maturation, providing an additional mechanism by which motile bacteria can initiate an acquired immune response. PMID- 12734365 TI - The induction of HIV Gag-specific CD8+ T cells in the spleen and gut-associated lymphoid tissue by parenteral or mucosal immunization with recombinant Listeria monocytogenes HIV Gag. AB - The induction of mucosal immunity is crucial in controlling viral replication during HIV infection. In this study we compare the ability of a recombinant Listeria monocytogenes that expresses and secretes the HIV Ag Gag to induce CD8(+) T cells against this Ag in the spleen, mesenteric lymph nodes, and Peyer's patches and the ability to provide effector Gag-specific CD8(+) T cells to the lamina propria after i.v., oral, or rectal administration of the vaccine. The levels of Ag-specific CD8(+)-activated T cells were measured ex vivo using intracellular cytokine staining for IFN-gamma and H-2K(d) Gag peptide tetramer staining. We found that all routes of immunization induced Gag-specific CD8(+) T cells in the spleen. After secondary infection, we observed substantial increases in splenic levels of CD8(+) T cells, and levels of Gag-specific cells were similar to those against listeriolysin O, the immunodominant Ag of L. monocytogenes. Both primary and secondary oral immunization resulted in abundant Gag-specific CD8(+)-activated T cells in the lamina propria that constituted approximately 35% of the CD8 compartment. However, significant levels of Gag and listeriolysin O-specific CD8(+) T cells were observed in mucosal lymphoid tissue only after two immunizations, perhaps because they had already entered the lamina propria compartment after a single immunization. In the context of HIV, a mucosally administered vaccine seems best calculated to prompt an immune response that is capable of preventing infection. The data presented in this report demonstrate that mucosally administered Listeria can prompt such a response and that booster doses can maintain this response. PMID- 12734366 TI - A single heteroclitic epitope determines cancer immunity after xenogeneic DNA immunization against a tumor differentiation antigen. AB - Successful active immunization against cancer requires induction of immunity against self or mutated self Ags. However, immunization against self Ags is difficult. Xenogeneic immunization with orthologous Ags induces cancer immunity. The present study evaluated the basis for immunity induced by active immunization against a melanoma differentiation Ag, gp100. Tumor rejection of melanoma was assessed after immunization with human gp100 (hgp100) DNA compared with mouse gp100 (mgp100). C57BL/6 mice immunized with xenogeneic full-length hgp100 DNA were protected against syngeneic melanoma challenge. In contrast, mice immunized with hgp100 DNA and given i.p. tolerizing doses of the hgp100 D(b)-restricted peptide, hgp100(25-33), were incapable of rejecting tumors. Furthermore, mice immunized with DNA constructs of hgp100 in which the hgp100(25-27) epitope was substituted with the weaker D(b)-binding epitope from mgp100 (mgp100(25-27)) or a mutated epitope unable to bind D(b) did not reject B16 melanoma. Mice immunized with a minigene construct of hgp100(25-33) rejected B16 melanoma, whereas mice immunized with the mgp100(25-33) minigene did not develop protective tumor immunity. In this model of xenogeneic DNA immunization, the presence of an hgp100 heteroclitic epitope with a higher affinity for MHC created by three amino acid (25 to 27) substitutions at predicted minor anchor residues was necessary and sufficient to induce protective tumor immunity in H-2(b) mice with melanoma. PMID- 12734367 TI - Monocyte-derived dendritic cells generated after a short-term culture with IFN alpha and granulocyte-macrophage colony-stimulating factor stimulate a potent Epstein-Barr virus-specific CD8+ T cell response. AB - Cellular immune responses are crucial for the control of EBV-associated lymphoproliferative diseases. To induce an anti-EBV cell-mediated immunity, we have used dendritic cells (DCs) generated by a 3-day culture of human CD14(+) monocytes in the presence of GM-CSF and type I IFN (IFN-DCs) and pulsed with peptides corresponding to CTL EBV epitopes. The functional activity of IFN-DCs was compared with that of APCs differentiated by culturing monocytes for 3 days with GM-CSF and IL-4 and indicated as IL-4-DCs. Stimulation of PBLs from EBV seropositive donors with EBV peptide-pulsed autologous IFN-DCs resulted in a stronger expansion of specific T lymphocytes producing IFN-gamma with respect to stimulation with peptide-loaded IL-4-DCs, as assessed by ELISPOT assays. When purified CD8(+) T cells were cocultured with EBV peptide-pulsed IFN-DCs or IL-4 DCs, significantly higher levels of specific cytotoxic activity were observed in CD8(+) T cell cultures stimulated with IFN-DCs. Injection of peptide-pulsed IFN DCs into SCID mice transplanted with autologous PBLs led to the recovery of a significantly greater number of EBV-specific human CD8(+) T cells from the spleen and the peritoneal cavity with respect to that recovered from mice injected with peptide-pulsed IL-4-DCs. Moreover, a significant delay in lymphoma development was observed when peptide-pulsed IFN-DCs were injected into SCID mice reconstituted with PBMCs endowed with a high capability of lymphoma induction, whereas injection of unpulsed IFN-DCs was ineffective. Our results indicate that IFN-DCs efficiently promote in vitro and in vivo the expansion of CD8(+) T lymphocytes acting as cytotoxic effectors against EBV-transformed cells. PMID- 12734368 TI - Cumulative Toll-like receptor activation in human macrophages treated with whole bacteria. AB - Purified components from bacteria selectively activate Toll-like receptors (TLR), leading to shared and unique responses in innate immune cells. Whole bacteria contain agonists for multiple TLR and induce a common macrophage activation program of transcription. It is not known, however, whether the stimulation of specific TLR by whole bacteria results in differential activation of the innate immune system. We evaluated gene expression data from human macrophages and found a unique gene expression profile induced by Gram-negative bacteria. In contrast, Gram-positive bacteria evoked few specific alterations in gene expression. LPS, a TLR4-specific ligand, was sufficient to elicit the distinct expression profile observed with Gram-negative bacteria. TLR4 activation regulated gene expression by both an IFN-dependent and an IFN-independent mechanism, illustrated by I-TAC and IL-12 p70, respectively. IL-12 p70 was produced by cells in whole blood exposed to Gram-negative bacteria, demonstrating faithful reproduction of the macrophage response in mixed populations of cells and identifying a potential diagnostic marker of infection. Our results show that the macrophage response to bacteria is dominated by the accumulated input from multiple TLR. For macrophages exposed to Gram-negative bacteria, gene expression changes encompass those induced by Gram-positive bacteria plus a distinct TLR4 response. This distinct TLR4 response may provide the basis to diagnose clinical Gram-negative infections. PMID- 12734369 TI - The lymphotoxin beta receptor is critically involved in controlling infections with the intracellular pathogens Mycobacterium tuberculosis and Listeria monocytogenes. AB - Containment of intracellularly viable microorganisms requires an intricate cooperation between macrophages and T cells, the most potent mediators known to date being IFN-gamma and TNF. To identify novel mechanisms involved in combating intracellular infections, experiments were performed in mice with selective defects in the lymphotoxin (LT)/LT beta R pathway. When mice deficient in LT alpha or LT beta were challenged intranasally with Mycobacterium tuberculosis, they showed a significant increase in bacterial loads in lungs and livers compared with wild-type mice, suggesting a role for LT alpha beta heterotrimers in resistance to infection. Indeed, mice deficient in the receptor for LT alpha(1)beta(2) heterotrimers (LT beta R-knockout (KO) mice) also had significantly higher numbers of M. tuberculosis in infected lungs and exhibited widespread pulmonary necrosis already by day 35 after intranasal infection. Furthermore, LT beta R-KO mice were dramatically more susceptible than wild-type mice to i.p. infection with Listeria monocytogenes. Compared with wild-type mice, LT beta R-KO mice had similar transcript levels of TNF and IFN-gamma and recruited similar numbers of CD3(+) T cells inside granulomatous lesions in M. tuberculosis-infected lungs. Flow cytometry revealed that the LT beta R is expressed on pulmonary macrophages obtained after digestion of M. tuberculosis infected lungs. LT beta R-KO mice showed delayed expression of inducible NO synthase protein in granuloma macrophages, implicating deficient macrophage activation as the most likely cause for enhanced susceptibility of these mice to intracellular infections. Since LIGHT-KO mice proved to be equally resistant to M. tuberculosis infection as wild-type mice, these data demonstrate that signaling of LT alpha(1)beta(2) heterotrimers via the LT beta R is an essential prerequisite for containment of intracellular pathogens. PMID- 12734370 TI - Macrophages restrict Pseudomonas aeruginosa growth, regulate polymorphonuclear neutrophil influx, and balance pro- and anti-inflammatory cytokines in BALB/c mice. AB - The role of macrophages in Pseudomonas aeruginosa corneal infection in susceptible (cornea perforates), C57BL/6 (B6) vs resistant (cornea heals), BALB/c mice was tested by depleting macrophages using subconjunctival injections of clodronate-containing liposomes before corneal infection. Both groups of inbred mice treated with clodronate-liposomes compared with PBS-liposomes (controls) exhibited more severe disease. In B6 mice, the cornea perforated and the eye became extremely shrunken, whereas in BALB/c mice, the cornea perforated rather than healed. The myeloperoxidase assay detected significantly more PMN in the cornea of both groups of mice treated with clodronate-liposomes vs PBS-liposomes. In independent experiments, ELISA analysis showed that protein levels for IL-1 beta, macrophage-inflammatory protein 2, and macrophage-inflammatory protein 1 alpha, all regulators of PMN chemotaxis, also were elevated in both groups of mice treated with clodronate-liposomes. Bacterial plate counts in B6 mice treated with clodronate-liposomes were unchanged at 3 days and were higher in control treated mice at 5 days postinfection (p.i.), whereas in BALB/c mice, bacterial load was significantly elevated in the cornea of mice treated with clodronate liposomes at both 3 and 5 days p.i. mRNA expression levels for pro (IFN-gamma and TNF-alpha)- and anti (IL-4 and IL-10)-inflammatory cytokines also were determined in BALB/c mice treated with clodronate-liposomes vs control-treated mice. Expression levels for IFN-gamma were significantly elevated in mice treated with clodronate-liposomes at 3 and 5 days p.i., while IL-10 levels (mRNA and protein) were reduced. These data provide evidence that macrophages control resistance to P. aeruginosa corneal infection through regulation of PMN number, bacterial killing and balancing pro- and anti-inflammatory cytokine levels. PMID- 12734371 TI - Neutrophilia in LFA-1-deficient mice confers resistance to listeriosis: possible contribution of granulocyte-colony-stimulating factor and IL-17. AB - LFA-1 (CD11a/CD18) plays a crucial role in various inflammatory responses. In this study, we show that LFA-1(-/-) mice are far more resistant to Listeria monocytogenes infection than LFA-1(+/-) mice. Consistent with this, we found the following: 1) the numbers of granulocytes infiltrating the liver were markedly higher in LFA-1(-/-) mice than in LFA-1(+/-) mice, 2) increased antilisterial resistance in LFA-1(-/-) mice was abrogated by depletion of granulocytes, and 3) the numbers of granulocytes in peripheral blood, and the serum levels of both G CSF and IL-17 were higher in LFA-1(-/-) mice than in LFA-1(+/-) mice. Neither spontaneous apoptosis nor survival of granulocytes from LFA-1(-/-) mice were affected by physiological concentrations of G-CSF. Our data suggest regulatory effects of LFA-1 on G-CSF and IL-17 secretion, and as a corollary on neutrophilia. Consequently, we conclude that increased resistance of LFA-1(-/-) mice to listeriosis is due to neutrophilia facilitating liver infiltration by granulocytes promptly after L. monocytogenes infection, although it is LFA-1 independent. PMID- 12734372 TI - Chemotactic factor-induced recruitment and activation of Tec family kinases in human neutrophils. II. Effects of LFM-A13, a specific Btk inhibitor. AB - Tyrosine phosphorylation events play major roles in the initiation and regulation of several functional responses of human neutrophils stimulated by chemotactic factors such as the bacterially derived tripeptide formylmethionyl-leucyl phenylalanine (fMet-Leu-Phe). However, the links between the G protein-coupled receptors, the activation of the tyrosine kinases, and the initiation of neutrophil functional responses remain unclear. In the present study we assessed the effects of a Btk inhibitor, leflunomide metabolite analog (LFM-A13), on neutrophils. LFM-A13 decreased the tyrosine phosphorylation induced by fMet-Leu Phe and inhibited the production of superoxide anions and the stimulation of adhesion, chemotaxis, and phospholipase D activity. We observed a decreased accumulation of phosphatidylinositol-3,4,5-trisphosphate in response to fMet-Leu Phe in LFM-A13-pretreated cells even though the inhibitor had no direct effect on the lipid kinase activity of the p110 gamma or p85/p110 phosphatidylinositol 3 kinases or on the activation of p110 gamma by fMet-Leu-Phe. The phosphorylation of Akt and of extracellular signal-regulated kinases 1/2 and p38 were similarly inhibited by LFM-A13. LFM-A13 also negatively affected the translocation of Rac 2, RhoA, ADP ribosylation factor-1, Tec, Bmx, and Btk induced by fMet-Leu-Phe. The results of this study provide evidence for an involvement of Btk and possibly other Tec kinase family members in the regulation of the functional responsiveness of human neutrophils and link these events, in part at least, to the modulation of levels of phosphatidylinositol-3,4,5-trisphosphate. PMID- 12734374 TI - C-C chemokine ligand 2/monocyte chemoattractant protein-1 directly inhibits NKT cell IL-4 production and is hepatoprotective in T cell-mediated hepatitis in the mouse. AB - T cell-mediated liver diseases are associated with elevated serum levels of C-C chemokine ligand 2 (CCL2)/monocyte chemoattractant protein-1 (MCP-1). However, the extent to which the actions of CCL2/MCP-1 contribute to the pathogenesis of T cell-mediated hepatitis remains incompletely understood. Con A-induced hepatitis is a liver-specific inflammation mediated by activated T cells and is driven by an up-regulation of the hepatic expression of TNF-alpha, IFN-gamma, and IL-4. The present study examined the role of CCL2/MCP-1 in the pathogenesis of T cell mediated hepatitis induced by Con A administration in the mouse. We demonstrate a novel hepatoprotective role for CCL2/MCP-1 during Con A-induced hepatitis, because CCL2/MCP-1 neutralization strikingly enhanced hepatic injury, both biochemically and histologically, after Con A administration. Furthermore, CCL2/MCP-1 neutralization was associated with a significant reduction in the hepatic levels of TNF-alpha and IFN-gamma, but with a significant increase in hepatic IL-4 levels. Moreover, IL-4 production and CCR2 expression by Con A stimulated CD3(+)NK1.1(+) T cells was significantly reduced by rMCP-1 treatment in vitro. In summary, we propose that CCL2/MCP-1 fulfills a novel anti inflammatory role in T cell-mediated hepatitis by inhibiting CD3(+)NK1.1(+) T cell-derived IL-4 production through direct stimulation of its specific receptor CCR2. These findings may have direct clinical relevance to T cell-mediated hepatitis. PMID- 12734373 TI - Duffy antigen facilitates movement of chemokine across the endothelium in vitro and promotes neutrophil transmigration in vitro and in vivo. AB - The Duffy Ag expressed on RBCs, capillaries, and postcapillary venular endothelial cells binds selective CXC and CC chemokines with high affinity. Cells transfected with the Duffy Ag internalize but do not degrade chemokine ligand. It has been proposed that Duffy Ag transports chemokines across the endothelium. We hypothesized that Duffy Ag participates in the movement of chemokines across the endothelium and, by doing so, modifies neutrophil transmigration. We found that the Duffy Ag transfected into human endothelial cells facilitates movement of the radiolabeled CXC chemokine, growth related oncogene-alpha/CXC chemokine ligand 1 (GRO-alpha/CXCL1), across an endothelial monolayer. In addition, neutrophil migration toward GRO-alpha/CXCL1 and IL-8 (IL-8/CXCL8) was enhanced across an endothelial monolayer expressing the Duffy Ag. Furthermore, GRO-alpha/CXCL1 stimulation of endothelial cells expressing the Duffy Ag did not affect gene expression by oligonucleotide microarray analysis. These in vitro observations are supported by the finding that IL-8/CXCL8-driven neutrophil recruitment into the lungs was markedly attenuated in transgenic mice lacking the Duffy Ag. We conclude that Duffy Ag has a role in enhancing leukocyte recruitment to sites of inflammation by facilitating movement of chemokines across the endothelium. PMID- 12734375 TI - Different modes of IL-10 and TGF-beta to inhibit cytokine-dependent IFN-gamma production: consequences for reversal of lipopolysaccharide desensitization. AB - LPS hyporesponsiveness is characterized by a diminished production of proinflammatory cytokines which can be caused by pretreatment with either LPS (=LPS desensitization) or the combination of the anti-inflammatory cytokines IL 10 and TGF-beta. However, the resulting hyporesponsive states differ regarding their reversibility by the IFN-gamma-inducing cytokine IL-12. Therefore, we aimed at studying the reasons for this differential IL-12 responsiveness of IFN-gamma producing cells and its consequences for LPS hyporesponsiveness in more detail. In an in vitro IL-12/IL-18 responsiveness model, we demonstrated that IL-10, if permanently present, does not directly inhibit IL-12/IL-18 responsiveness in T/NK cells but indirectly interferes with IFN-gamma production in the presence of monocytes. In contrast, TGF-beta acted directly on IFN-gamma-producing cells by interfering with IL-12/IL-18 responsiveness. After removal of IL-10 but not of TGF-beta, LPS hyporesponsiveness can be reverted by IL-12/IL-18. Consequently, the addition of recombinant TGF-beta during LPS desensitization rendered PBMCs hyporesponsive to a reversal by IL-12/IL-18. Our data suggest that the persistence of IL-10 and the presence of TGF-beta determine the level of IFN gamma inhibition and may result in different functional phenotypes of LPS desensitization and LPS hyporesponsiveness in vitro and in vivo. PMID- 12734376 TI - Selective roles for Toll-like receptor (TLR)2 and TLR4 in the regulation of neutrophil activation and life span. AB - Neutrophil responses to commercial LPS, a dual Toll-like receptor (TLR)2 and TLR4 activator, are regulated by TLR expression, but are amplified by contaminating monocytes in routine cell preparations. Therefore, we investigated the individual roles of TLR2 and TLR4 in highly purified, monocyte-depleted neutrophil preparations, using selective ligands (TLR2, Pam(3)CysSerLys(4) and Staphylococcus aureus peptidoglycan; TLR4, purified LPS). Activation of either TLR2 or TLR4 caused changes in adhesion molecule expression, respiratory burst (alone, and synergistically with fMLP), and IL-8 generation, which was, in part, dependent upon p38 mitogen-activated protein kinase signaling. Neutrophils also responded to Pam(3)CysSerLys(4) and purified LPS with down-regulation of the chemokine receptor CXCR2 and, to a lesser extent, down-regulation of CXCR1. TLR4 was the principal regulator of neutrophil survival, and TLR2 signals showed relatively less efficacy in preventing constitutive apoptosis over short time courses. TLR4-mediated neutrophil survival depended upon signaling via NF-kappa B and mitogen-activated protein kinase cascades. Prolonged neutrophil survival required both TLR4 activation and the presence of monocytes. TLR4 activation of monocytes was associated with the release of neutrophil survival factors, which was not evident with TLR2 activation, and TLR2 activation in monocyte/neutrophil cocultures did not prevent late neutrophil apoptosis. Thus, TLRs are important regulators of neutrophil activation and survival, with distinct and separate roles for TLR2 and TLR4 in neutrophil responses. TLR4 signaling presents itself as a pharmacological target that may allow therapeutic modulation of neutrophil survival by direct and indirect mechanisms at sites of inflammation. PMID- 12734377 TI - Calcium-independent phospholipase A2 is required for lysozyme secretion in U937 promonocytes. AB - As a part of their surveillance functions in the immune system, monocytes/macrophages secrete large amounts of the bactericidal enzyme lysozyme to the extracellular medium. We report here that lysozyme secretion in activated U937 promonocytes depends on a functional calcium-independent phospholipase A(2) (iPLA(2)). Inhibition of the enzyme by bromoenol lactone or by treatment with a specific antisense oligonucleotide results in a diminished capacity of the cells to secrete lysozyme to the extracellular medium. Calcium-independent PLA(2) is largely responsible for the maintenance of the steady state of lysophosphatidylcholine (lysoPC) levels within the cells, as manifested by the marked decrease in the levels of this metabolite in cells deficient in iPLA(2) activity. Reconstitution experiments reveal that lysoPC efficiently restores lysozyme secretion in iPLA(2)-deficient cells, whereas other lysophospholipids, including lysophosphatidic acid, lysophosphatidylserine, and lysophosphatidylethanolamine, are without effect. Arachidonic acid mobilization in activated U937 cells is under control of cytosolic phospholipase A(2) (cPLA(2)). Selective inhibition of cPLA(2) results in a complete abrogation of the arachidonate mobilization response, but has no effect on lysozyme secretion. These results identify iPLA(2)-mediated lysoPC production as a necessary component of the molecular machinery leading to lysozyme secretion in U937 cells and rule out a role for cPLA(2) in the response. Collectively, the results demonstrate distinct roles in inflammatory cell signaling for these two intracellular phospholipases. PMID- 12734378 TI - Amyloid peptide-induced cytokine and chemokine expression in THP-1 monocytes is blocked by small inhibitory RNA duplexes for early growth response-1 messenger RNA. AB - In Alzheimer's disease (AD) one finds increased deposition of A beta and also an increased presence of monocytes/macrophages in the vessel wall and activated microglial cells in the brain. AD patients show increased levels of proinflammatory cytokines by activated microglia. Here we used a human monocytic THP-1 cell line as a model for microglia to delineate the cellular signaling mechanism involved in amyloid peptides (A beta(1-40) and A beta(1-42))-induced expression of inflammatory cytokines and chemokines. We observed that A beta peptides at physiological concentrations (125 nM) increased mRNA expression of cytokines (TNF-alpha, and IL-1 beta) and chemokines (monocyte chemoattractant protein-1 (MCP-1), IL-8, and macrophage inflammatory protein-1 beta (MIP-1 beta)). The cellular signaling involved activation of c-Raf, extracellular signal regulated kinase-1 (ERK-1)/ERK-2, and c-Jun N-terminal kinase, but not p38 mitogen-activated protein kinase. This is further supported by the data showing that A beta causes phosphorylation of ERK-1/ERK-2, which, in turn, activates Elk 1. Furthermore, A beta mediated a time-dependent increase in DNA binding activity of early growth response-1 (Egr-1) and AP-1, but not of NF-kappa B and CREB. Moreover, A beta-induced Egr-1 DNA binding activity was reduced >60% in THP-1 cells transfected with small interfering RNA duplexes for Egr-1 mRNA. We show that A beta-induced expression of TNF-alpha, IL-1 beta, MCP-1, IL-8, and MIP-1 beta was abrogated in Egr-1 small inhibitory RNA-transfected cells. Our results indicate that A beta-induced expression of cytokines (TNF-alpha and IL-1 beta) and chemokines (MCP-1, IL-8, and MIP-1 beta) in THP-1 monocytes involves activation of ERK-1/ERK-2 and downstream activation of Egr-1. The inhibition of Egr-1 by Egr-1 small inhibitory RNA may represent a potential therapeutic target to ameliorate the inflammation and progression of AD. PMID- 12734379 TI - Peroxisome proliferator-activated receptor gamma inhibits the migration of dendritic cells: consequences for the immune response. AB - The migration of dendritic cells (DCs) from the epithelia to the lymphoid organs represents a tightly regulated multistep event involved in the induction of the immune response. In this process fatty acid derivatives positively and negatively regulate DC emigration. In the present study we investigated whether activation of peroxisome proliferator-activated receptors (PPARs), a family of nuclear receptors activated by naturally occurring derivatives of arachidonic acid, could control DC migration from the peripheral sites of Ag capture to the draining lymph nodes (DLNs). First, we show that murine epidermal Langerhans cells (LCs) express PPAR gamma, but not PPAR alpha, mRNA, and protein. Using an experimental murine model of LC migration induced by TNF-alpha, we show that the highly potent PPAR gamma agonist rosiglitazone specifically impairs the departure of LCs from the epidermis. In a model of contact allergen-induced LC migration, PPAR gamma activation not only impedes LC emigration, and their subsequent accumulation as DCs in the DLNs, but also dramatically prevents the contact hypersensitivity responses after challenge. Finally, after intratracheal sensitization with an FITC-conjugated Ag, PPAR gamma activation inhibits the migration of DCs from the airway mucosa to the thoracic LNs and also profoundly reduces the priming of Ag specific T lymphocytes in the DLNs. Our results suggest a novel regulatory pathway via PPAR gamma for DC migration from epithelia that could contribute to the initiation of immune responses. PMID- 12734380 TI - Akt phosphorylates p47phox and mediates respiratory burst activity in human neutrophils. AB - Respiratory burst activity and phosphorylation of an NADPH oxidase component, p47(phox), during neutrophil stimulation are mediated by phosphatidylinositol 3 kinase (PI-3K) activation. Products of PI-3K activate several kinases, including the serine/threonine kinase Akt. The present study examined the ability of Akt to regulate neutrophil respiratory burst activity and to interact with and phosphorylate p47(phox). Inhibition of Akt activity in human neutrophils by an inhibitory peptide significantly attenuated fMLP-stimulated, but not PMA stimulated, superoxide release. Akt inhibitory peptide also inhibited hydrogen peroxide generation stimulated by bacterial phagocytosis. A direct interaction between p47(phox) and Akt was shown by the ability of GST-p47(phox) to precipitate recombinant Akt and to precipitate Akt from neutrophil lysates. Active recombinant Akt phosphorylated recombinant p47(phox) in vitro, as shown by (32)P incorporation, by a mobility shift change detected by two-dimensional gel electrophoresis, and by immunoblotting with phospho-Akt substrate Ab. Mutation analysis indicated that 2 aa residues, Ser(304) and Ser(328), were phosphorylated by Akt. Inhibition of Akt activity also inhibited fMLP-stimulated neutrophil chemotaxis. We propose that Akt mediates PI-3K-dependent p47(phox) phosphorylation, which contributes to respiratory burst activity in human neutrophils. PMID- 12734381 TI - Expression and production of the CXC chemokine growth-related oncogene-alpha by human eosinophils. AB - Eosinophils are seen together with neutrophils at sites of inflammation. However, their roles are not clear. In addition, eosinophils infiltrate tumor tissue in some neoplastic diseases. In this study, we show that large amounts of the neutrophil-activating CXC chemokine growth-related oncogene (GRO)-alpha can be produced by human eosinophils. Eosinophils showed presence of preformed GRO-alpha in the crystalloid-containing specific granules (190 pg/2 x 10(6) cells). During incubation, a strong increase in GRO-alpha gene expression was seen. At a low cell density, addition of TNF-alpha or IL-1 beta increased the production of GRO alpha in eosinophils, which was not the case at a higher cell density. Eosinophils can produce TNF-alpha themselves, and neutralizing Abs against TNF alpha significantly inhibited GRO-alpha production. This suggests that autocrine and paracrine effects from TNF-alpha can be important when up-regulating GRO alpha gene expression. In contrast, IFN-gamma, a prototypic Th1-cytokine, down regulated expression of GRO-alpha. This may be important during resolution of inflammation but also suggests different roles for eosinophils depending on the inflammatory context. Tumor-infiltrating eosinophils in Hodgkin's disease of the nodular sclerosing type are associated with a poor prognosis. Eosinophils from such tumor tissue showed an abundant expression of GRO-alpha. The GRO-alpha receptor CXCR2 was also detected in tumor tissue, proposing interactions between eosinophils and the tumor. Our findings suggest that eosinophils can promote inflammation through recruitment of CXCR2-bearing cells. In addition, this feature of the eosinophils indicates a role for these cells in the biology of certain tumors. PMID- 12734383 TI - Immunization with a xenobiotic 6-bromohexanoate bovine serum albumin conjugate induces antimitochondrial antibodies. AB - The E2 subunit of pyruvate dehydrogenase complex (PDC-E2) is the major autoantigen recognized by antimitochondrial Abs (AMA) in primary biliary cirrhosis (PBC). Recently, we replaced the lipoic acid moiety of PDC-E2 with a battery of synthetic structures designed to mimic a xenobiotically modified lipoyl hapten on a 12-aa peptide that was found within the immunodominant autoepitope of PDC-E2 and demonstrated that AMA in PBC reacted against several organic modified mimotopes as well as, or sometimes significantly better than, the native lipoyl domain. Based on this data, we immunized rabbits with one such xenobiotic organic compound, 6-bromohexanoate, coupled to BSA. One hundred percent of immunized rabbits developed AMA that have each and every characteristic of human AMAs with reactivity against PDC-E2, E2 subunit of branched chain 2-oxo-acid dehydrogenase, and E2 subunit of 2-oxoglutarate dehydrogenase complex. The rabbit AMA also inhibited enzymatic function of PDC-E2 and, importantly, binds to peptide sequences not present in the xenobiotic carrier immunogen. In contrast, BSA-immunized controls did not produce such activity. Our observation that animals immunized with a xenobiotic BSA complex produce autoantibodies that react not only with the xenobiotic, but also with mitochondrial autoantigens recognized by autoimmune PBC sera, suggests that environmental xenobiotic agents can be a risk factor for the induction of PBC. PMID- 12734382 TI - Hybrids of dendritic cells and tumor cells generated by electrofusion simultaneously present immunodominant epitopes from multiple human tumor associated antigens in the context of MHC class I and class II molecules. AB - Hybrid cells generated by fusing dendritic cells with tumor cells (DC-TC) are currently being evaluated as cancer vaccines in preclinical models and human immunization trials. In this study, we evaluated the production of human DC-TC hybrids using an electrofusion protocol previously defined for murine cells. Human DCs were electrically fused with allogeneic melanoma cells (888mel) and were subsequently analyzed for coexpression of unique DC and TC markers using FACS and fluorescence microscopy. Dually fluorescent cells were clearly observed using both techniques after staining with Abs against distinct surface molecules suggesting that true cell fusion had occurred. We also evaluated the ability of human DC-TC hybrids to present tumor-associated epitopes in the context of both MHC class I and class II molecules. Allogeneic DCs expressing HLA-A*0201, HLA-DR beta 1*0401, and HLA-DR beta 1*0701 were fused with 888mel cells that do not express any of these MHC molecules, but do express multiple melanoma-associated Ags. DC-888mel hybrids efficiently presented HLA-A*0201-restricted epitopes from the melanoma Ags MART-1, gp100, tyrosinase, and tyrosinase-related protein 2 as evaluated by specific cytokine secretion from six distinct CTL lines. In contrast, DCs could not cross-present MHC class I-restricted epitopes after exogenously loading with gp100 protein. DC-888mel hybrids also presented HLA-DR beta 1*0401- and HLA-DR beta 1*0701-restricted peptides from gp100 to CD4(+) T cell populations. Therefore, fusions of DCs and tumor cells express both MHC class I- and class II-restricted tumor-associated epitopes and may be useful for the induction of tumor-reactive CD8(+) and CD4(+) T cells in vitro and in human vaccination trials. PMID- 12734384 TI - Angiotensin type 2 receptor (AT2R): a challenging twin. AB - Angiotensin II (AngII) regulates such physiological responses as salt and water balance, blood pressure, and vascular tone, and thus plays a critical role in the pathogenesis of diabetes, hypertension, myocardial infarction, congestive heart failure, and stroke. These effects are mediated through at least three receptors: AT1R, AT2R, and AT4R, which are expressed under different developmental, tissue specific, and disease-specific conditions and which couple to distinct effector pathways. Signaling through the AT1R, a classical G protein-coupled receptor, has been extensively studied and is well understood. Less is known about signaling through the AT2R, which often antagonizes the effects of signaling through the AT1R, but intriguing data are beginning to emerge concerning the signaling strategies and pathways that the AT2R employs. PMID- 12734386 TI - Evolutionary relationships of the Tas2r receptor gene families in mouse and human. AB - The early molecular events in the perception of bitter taste start with the binding of specific water-soluble molecules to G protein-coupled receptors (GPCRs) encoded by the Tas2r family of taste receptor genes. The identification of the complete TAS2R receptor family repertoire in mouse and a comparative study of the Tas2r gene families in mouse and human might help to better understand bitter taste perception. We have identified, cloned, and characterized 13 new mouse Tas2r sequences, 9 of which encode putative functional bitter taste receptors. The encoded proteins are between 293 and 333 amino acids long and share between 18% and 54% sequence identity with other mouse TAS2R proteins. Including the 13 sequences identified, the mouse Tas2r family contains approximately 30% more genes and 60% fewer pseudogenes than the human TAS2R family. Sequence and phylogenetic analyses of the proteins encoded by all mouse and human Tas2r genes indicate that TAS2R proteins present a lower degree of sequence conservation in mouse than in human and suggest a classification in five groups that may reflect a specialization in their functional activity to detect bitter compounds. Tas2r genes are organized in clusters in both mouse and human genomes, and an analysis of these clusters and phylogenetic analyses indicates that the five TAS2R protein groups were present prior to the divergence of the primate and rodent lineages. However, differences in subsequent evolutionary processes, including local duplications, interchromosomal duplications, divergence, and deletions, gave rise to species-specific sequences and shaped the diversity of the current TAS2R receptor families during mouse and human evolution. PMID- 12734385 TI - Epidermal growth factor receptor signaling intensity determines intracellular protein interactions, ubiquitination, and internalization. AB - Ligand activation of the epidermal growth factor receptor (EGFR) causes the binding of Cbls, which leads to EGFR polyubiquitination and internalization through endophilin complexes that contain the adaptor protein SH3-domain encoding, expressed in tumorigenic astrocytes/Cbl-interacting protein of 85 kDa/regulator of ubiquitous kinase (SETA/CIN85/Ruk). In cells grown at high density, high levels of SETA interfered in the recruitment of Casitas B-lineage (Cbl) proteins to the EGFR and reduced its polyubiquitination, suggesting that SETA has a regulatory function in the formation of the EGFR-Cbl-endophilin complex and in EGFR down-regulation. In a situation where there is EGFR signaling but no internalization or down-regulation, as is the case with the EGFR with exons 2-7 deleted (DeltaEGFR) oncogene, these proteins were absent altogether. By using mAb 806, which recognizes an EGFR-activation state and preferentially immunoprecipitates DeltaEGFR, we show that DeltaEGFR did not interact with Cbls, SETA, or endophilin A1, providing a mechanistic explanation for its lack of internalization. As would be expected by the absence of Cbl proteins in the DeltaEGFR complex, the mutant receptor was also not polyubiquitinated. The intracellular C terminus and tyrosine autophosphorylation pattern of DeltaEGFR are similar to wild-type EGFR, but it signals at a lower intensity as determined by levels of EGFR phosphotyrosine. To test the implication that the lack of interaction with the Cbl-SETA-endophilin complex is because of differences in signal intensity, EGFR-expressing cells were treated with tyrphostin AG1478 EGFR inhibitor. Attenuation of wild-type EGFR signal to levels similar to that found in DeltaEGFR resulted in the dissociation of SETA and Cbl proteins and a concomitant attenuation of receptor internalization. PMID- 12734387 TI - Selective enrichment of thiophosphorylated polypeptides as a tool for the analysis of protein phosphorylation. AB - A chemoselective alkylation method is described for the isolation and subsequent identification of thiophosphorylated peptides/proteins. The method involves thiophosphorylation of proteins using adenosine 5'-O-(thiotriphosphate) (ATPgammaS) followed by selective in situ alkylation of the newly thiophosphorylated proteins resulting in a stable covalent bond. The chemoselective alkylation exploits the relatively high nucleophilicity at low pH of the sulfur in thiophosphate residues, whereas the nucleophilicities of phosphates, amines, and other functionality of amino acids are negligible or significantly suppressed. Modified alkylation reagents linked to biotin or solid supports (e.g. glass or Sepharose beads) with or without a photocleavable linker facilitate the isolation of the thiophosphorylated peptide/proteins. This approach is demonstrated through the localization of phosphorylation sites on myosin regulatory light chain. We anticipate that this technique will be useful for isolation and subsequent identification of newly thiophosphorylated proteins, produced either in vivo or in vitro, thus facilitating the dissection of protein phosphorylation networks. PMID- 12734388 TI - Mediation of corticotropin releasing factor type 1 receptor phosphorylation and desensitization by protein kinase C: a possible role in stress adaptation. AB - Protein kinase C (PKC)-mediated desensitization of the corticotropin releasing factor type 1 (CRF1) receptor was investigated in human retinoblastoma Y79 and transfected COS-7 cells. Because stimulation of Y79 cells with CRF resulted in large ( approximately 30-fold) increases in intracellular cAMP accumulation without changing inositol phosphate levels, the CRF1 receptor expressed in retinoblastoma cells couples to Gs, but not to Gq, and predominantly signals via the protein kinase A cascade. Direct activation of PKC by treatment with the phorbol ester phorbol 12-myristate 13-acetate (PMA) or 1,2-dioctanoyl-sn-glycerol (DOG) desensitized CRF1 receptors in Y79 cells, reducing the maximum for CRF- (but not forskolin)-stimulated cAMP accumulation by 56.3 +/- 1.2% and 40.4 +/- 2.1%, respectively (p < 0.001). Pretreating Y79 cells with the PKC inhibitor bisindolylmaleimide I (BIM) markedly inhibited PMA's desensitizing action on CRF stimulated cAMP accumulation, but did not affect homologous CRF1 receptor desensitization. Retinoblastoma cells were found to express PKCalpha, betaI, betaII, delta, lambda, and RACK1. When alpha and beta isoforms of PKC were down regulated 80 to 90% by a 48-h PMA exposure, PMA-induced CRF1 receptor desensitization was abolished. In transfected COS-7 cells the magnitude of CRF1 receptor phosphorylation after a 5-min exposure to PMA was 2.32 +/- 0.21-fold greater compared with the basal level. Pretreating COS-7 cells with BIM abolished PMA-induced CRF1 receptor phosphorylation. These studies demonstrate that protein kinase C (possibly alpha and beta isoforms) has an important role in the phosphorylation and heterologous desensitization of the CRF1 receptor. PMID- 12734389 TI - Regulation of septo-hippocampal activity by 5-hydroxytryptamine(2C) receptors. AB - It is established that the serotonin system modulates hippocampal functions by regulating neuronal activity of both the medial septum and hippocampus. Inhibition of serotonin neurons leads to theta oscillation of septal neurons and theta wave activity in the hippocampus, indicating a tonic regulation of the septo-hippocampal system by serotonin neurons. Because the postsynaptic 5 hydroxytryptamine (5-HT) receptor subtypes mediating this tonic inhibition have not been identified, a putative role of 5-HT2C receptors has been evaluated in the present study. Extracellular single units were recorded from the medial septum/vertical limb of diagonal band (MS/DBv) and hippocampal CA1 or dentate gyrus with simultaneous hippocampal EEG recordings from anesthetized rats. Intravenous administration of 5-HT2C receptor agonists 1-(3 chlorophenyl)piperazine dihydrochloride (m-CPP) and [S]-2-(chloro-5-fluoro-indol 1-yl)-1-methyl-ethylamine fumarate (Ro 60-0175) dose dependently inhibited firing activity most of the recorded MS/DBv neurons and abolished theta oscillation in all tested MS/DBv and hippocampal neurons. Parallel to inhibition of theta oscillation of MS/DBv neurons, hippocampal EEG activity was desynchronized and its power spectrum was shifted to lower frequencies. The selective 5-HT2C receptor antagonist 6-chloro-5-methyl-1-[2-(2-methylpyridyl-3-oxy)-pyrid-5-yl carbomyl] indoline (SB-242084) [but not the 5-HT2B antagonist 2-amino-4-(4 fluoronaphth-1-yl)-6-isopropyl-pyrimidine (RS-127445) or 5-HT2A antagonist R-(+) alpha-(2,3-dimethoxyphenyl)-1-[2-(4-fluorophenyl)-ethyl]-4-piperidinemethanol (MDL-100907)] reversed the action of 5-HT2C receptor agonists. Furthermore, in control rats 5-HT2C receptor antagonists [SB-242084 and 5-methyl-1-(3-pyridil carbamoyl)-1,2,3,5-tetrahydropyr-rolo[2,3-f]indole hydrochloride (SB-206553)] induced or enhanced theta oscillation in MS/DBv and hippocampal neurons and theta wave activity of the hippocampus. These findings provide evidence for a tonic regulation of the activity and theta oscillation of the septo-hippocampal system via 5-HT2C receptors in the anesthetized rat, indicating that pharmacological interactions with these receptors could modulate various physiological and pathological processes associated with limbic theta activity. PMID- 12734390 TI - Identification of residues that confer alpha-conotoxin-PnIA sensitivity on the alpha 3 subunit of neuronal nicotinic acetylcholine receptors. AB - Neuronal nicotinic receptors composed of the alpha3 and beta2 subunits are at least 1000-fold more sensitive to blockade by alpha-conotoxin-PnIA than are alpha2beta2 receptors. A series of chimeric subunits, formed from portions of alpha2 and alpha3, were coexpressed with beta2 in Xenopus oocytes and tested for toxin sensitivity. We found determinants of toxin sensitivity to be widely distributed in the extracellular domain of alpha3. Analysis of receptors formed by a series of mutant alpha3 subunits, in which residues that differ between alpha3 and alpha2 were changed from what occurs in alpha3 to what occurs in alpha2, allowed identification of three determinants of alpha-conotoxin-PnIA sensitivity: proline 182, isoleucine 188, and glutamine 198. Comparison with determinants of alpha-conotoxin-MII and kappa-bungarotoxin sensitivity on the alpha3 subunit revealed overlapping, but distinct, arrays of determinants for each of these three toxins. When tested against an EC50 concentration of acetylcholine, the IC50 for alpha-conotoxin-PnIA blockade was 25 +/- 4 nM for alpha3beta2, 84 +/- 7 nM for alpha3P182Tbeta2, 700 +/- 92 nM for alpha3I188Kbeta2, and 870 +/- 61 nM for alpha3Q198Pbeta2. To examine the location of these residues within the receptor structure, we generated a homology model of the alpha3beta2 receptor extracellular domain using the structure of the acetylcholine binding protein as a template. All three residues are located on the C-loop of the alpha3 subunit, with isoleucine 188 nearest the acetylcholine binding pocket. PMID- 12734391 TI - Nicotinic acetylcholine receptor-mediated neuroprotection by donepezil against glutamate neurotoxicity in rat cortical neurons. AB - Donepezil is a potent and selective acetylcholinesterase (AChE) inhibitor developed for the treatment of Alzheimer's disease. To elucidate whether donepezil shows neuroprotective action in addition to amelioration of cognitive deficits, we examined the effects of donepezil on glutamate-induced neurotoxicity using primary cultures of rat cortical neurons. A 10-min exposure of cultures to glutamate followed by a 1-h incubation with glutamate-free medium caused a marked loss of viability, as determined by Trypan blue exclusion. Glutamate neurotoxicity was prevented by 24-h pretreatment of donepezil in a concentration dependent manner. Among AChE inhibitors examined, donepezil and certain AChE inhibitors such as tacrine and galanthamine showed potent neuroprotective action, although physostigmine did not affect glutamate neurotoxicity. Neuroprotective action of donepezil was antagonized by mecamylamine, a nicotinic acetylcholine receptor (nAChR) antagonist, but not by scopolamine, a muscarinic acetylcholine receptor antagonist. Furthermore, both dihydro-beta-erythroidine, an alpha4beta2 neuronal nAChR antagonist, and methyllycaconitine, an alpha7-nAChR antagonist, each also significantly antagonized the effect of donepezil. Next, we examined the effects of donepezil on glutamate-induced apoptosis. Exposure of 100 microM glutamate to cortical neurons for 24 h induced apoptotic neuronal death and nuclear fragmentation. Donepezil for 24 h before and 24 h during glutamate exposure prevented nuclear fragmentation and glutamate-induced apoptosis. These results suggest that donepezil not only protects cortical neurons against glutamate neurotoxicity via alpha4beta2- and alpha7-nAChRs but also prevents apoptotic neuronal death. PMID- 12734392 TI - Ethanol inhibits alpha-amino-3-hydyroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor function in central nervous system neurons by stabilizing desensitization. AB - Ethanol actions on alpha-amino-3-hydyroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptors were studied using voltage-clamp recordings from mouse cortical and hippocampal neurons. During whole-cell recordings ethanol (EtOH) inhibited AMPA receptor-mediated currents in a dose-dependent manner at concentrations from 10 to 500 mM. The steady-state component of AMPA-activated current was more sensitive to EtOH than the peak component. To examine the effect of EtOH on a well resolved peak current component, patches were excised from cultured cortical neurons, to which AMPA and EtOH were applied using a piezoelectric solution application system. Under this condition, the peak current was not inhibited significantly by EtOH. To further study possible mechanisms of EtOH inhibition, kainate and AMPA were used to evoke currents in the absence and presence of cyclothiazide. Ethanol inhibition was stronger when receptors were activated by low than high kainate concentrations. Cyclothiazide reduced inhibition by EtOH regardless of the agonist used to activate the receptor. Finally, EtOH inhibition was reduced in a point mutated (L497Y) GluRAi receptor that lacks desensitization. These findings suggest that EtOH inhibits AMPA receptors by stabilizing the desensitized state. Our results can explain some of the variation observed in EtOH inhibition in previous studies, and support the idea that physiologically relevant concentrations of EtOH can have a strong effect on AMPA receptor function. PMID- 12734393 TI - Inhibition of microglial activation attenuates the development but not existing hypersensitivity in a rat model of neuropathy. AB - Microglia, the intrinsic macrophages of the central nervous system, have previously been shown to be activated in the spinal cord in several rat mononeuropathy models. Activation of microglia and subsequent release of proinflammatory cytokines are known to play a role in inducing a behavioral hypersensitive state (hyperalgesia and allodynia) in these animals. The present study was undertaken to determine whether minocycline, an inhibitor of microglial activation, could attenuate both the development and existing mechanical allodynia and hyperalgesia in an L5 spinal nerve transection model of neuropathic pain. In a preventive paradigm (to study the effect on the development of hypersensitive behaviors), minocycline (10, 20, or 40 mg/kg intraperitoneally) was administered daily, beginning 1 h before nerve transection. This regimen produced a decrease in mechanical hyperalgesia and allodynia, with a maximum inhibitory effect observed at the dose of 20 and 40 mg/kg. The attenuation of the development of hyperalgesia and allodynia by minocycline was associated with an inhibitory action on microglial activation and suppression of proinflammatory cytokines at the L5 lumbar spinal cord of the nerveinjured animals. The effect of minocycline on existing allodynia was examined after its intraperitoneal administration initiated on day 5 post-L5 nerve transection. Although the postinjury administration of minocycline significantly inhibited microglial activation in neuropathic rats, it failed to attenuate existing hyperalgesia and allodynia. These data demonstrate that inhibition of microglial activation attenuated the development of behavioral hypersensitivity in a rat model of neuropathic pain but had no effect on the treatment of existing mechanical allodynia and hyperalgesia. PMID- 12734394 TI - Glycoprotein IIb/IIIa receptor antagonist (2S)-2-[(2-Naphthyl-sulfonyl)amino]-3 [[2-([4-(4-piperidinyl)-2-[2-(4-piperidinyl)ethyl] butanoyl]amino)acetyl]amino]propanoic acid dihydrochloride (CRL42796), in combination with aspirin and/or enoxaparin, prevents coronary artery rethrombosis after successful thrombolytic treatment by recombinant tissue plasminogen activator. AB - The antithrombotic effect of the glycoprotein IIb/IIIa (GPIIb/IIIa) antagonist (2S)-2-[(2-naphthyl-sulfonyl)amino]-3-[[2-([4-(4-piperidinyl)-2-[2-(4 piperidinyl)ethyl] butanoyl]amino)acetyl]amino] propanoic acid dihydrochloride (CRL42796), administered alone, or in combination with aspirin, and/or enoxaparin, was examined in a canine left circumflex (LCX) coronary artery rethrombosis model. The electrolytic induction of arterial thrombosis was followed by intracoronary recombinant tissue plasminogen activator administration to achieve thrombolysis, and the adjunctive therapy was initiated 15 min earlier and maintained for 4 h. Thirty-five purpose-bred beagle dogs were randomized to receive one of the following treatments: group 0 (n = 6, placebo); group 1 (n = 6, CRL42796 15 microg/kg i.v. loading dose followed by 0.31 microg/kg/min i.v. infusion), group 2 (n = 6, aspirin 7 mg/kg, administered orally, at -47, -23, -17 h before entry into the experimental protocol); group 3 (n = 6, aspirin + CRL42796); group 4 (n = 6, aspirin + enoxaparin 0.6 microg/kg i.v. loading dose followed by 6.0 microg/kg/min i.v. infusion); and group 5 (n = 5, aspirin + CRL42796 + enoxaparin). The incidence of LCX reocclusion was as follows: group 0, 6/6; group 1, 3/6; group 2, 5/6; group 3, 2/6; group 4, 2/6; and group 5, 0/5. Aspirin pretreatment increased the tongue-bleeding time, whereas the addition of CRL42796 or enoxaparin did not prolong bleeding time to a further degree. However, the combination of the three drugs did increase bleeding time significantly, from 173.9 +/- 19.8 to 620.0 +/- 98.7 s. In conclusion, low-dose CRL42796 together with aspirin and enoxaparin prevented coronary artery rethrombosis, although bleeding time was prolonged. The latter may be of concern in the clinical use of combination therapy. PMID- 12734395 TI - Forward EphB4 signaling in endothelial cells controls cellular repulsion and segregation from ephrinB2 positive cells. AB - Contact-dependent interactions between endothelial cells (ECs), as well as between ECs and mural cells, play a key role in the formation of a regular vascular system and the assembly of the vessel wall. Recent studies have identified ephrinB2 and EphB4 as markers and makers of arteriovenous differentiation during vascular development. On the basis of these findings, we hypothesized that Ephephrin interactions in the vascular system mediate distinct propulsive and repulsive effector functions that provide guidance signals for the proper spatial organization of the developing vasculature. Utilizing a set of specialized endothelial differentiation and angiogenesis assays, the present study was aimed at studying vascular morphogenic functions of endothelial EphB4 and ephrinB2 activation. EphrinB2-Fc acts anti-adhesively and induces detachment of ECs, as demonstrated by (1) inhibition of adhesion to ephrinB2-Fc-coated culture dishes, (2) detachment of ECs grown as differentiated 3D spheroids, and (3) endothelial denudation of explanted fragments of umbilical vein. Conversely, soluble ephrinB2-Fc inhibits lateral cell migration, vascular endothelial growth factor (VEGF) gradient-driven chemotaxis, capillary-like network formation and sprouting angiogenesis. In turn, soluble EphB4-Fc is pro-adhesive and stimulates EC migration and sprouting angiogenesis. EphrinB2-mediated repulsive signals are transduced by EphB4, as demonstrated by EphrinB2-Fc inhibition of sprouting angiogenesis of constitutively EphB4-overexpressing ECs. Confrontation experiments of EphB4-overexpressing ECs with ECs overexpressing full-length or truncated ephrinB2 that lacks the cytoplasmic catalytic domain demonstrated that forward EphB4 signaling with EphB4 tyrosine phosphorylation restricts intermingling of cells and supports cellular segregation. Taken together, these data identify distinct propulsive and repulsive effector functions of endothelial ephrinB2 and EphB4 that mediate spatial positional signals during angiogenesis and vessel assembly. PMID- 12734396 TI - The lamina-associated polypeptide 2 (LAP2) isoforms beta, gamma and omega of zebrafish: developmental expression and behavior during the cell cycle. AB - Zebrafish lamina-associated polypeptides 2 (ZLAP2) beta, gamma and omega have in common an N-terminal region with a LEM domain, and in the C-terminal half of the molecule a lamina binding domain and a membrane spanning sequence. The maternally synthesized omega is the largest isoform and the only LAP2 present in the rapidly dividing embryonic cells up to the gastrula stage. ZLAP2omega levels decrease during development, concomitant with the increase of the somatic isoforms ZLAP2beta and gamma. In somatic zebrafish cells ZLAP2gamma is the predominant isoform, whereas only small amounts of ZLAP2beta are present. During early embryonic development, ZLAP2omega becomes associated with mitotic chromosomes before anaphase. The surface of these chromosomes is decorated with vesicles, and each chromosome assembles its own nuclear envelope at the end of mitosis (karyomere formation). Ectopically expressed ZLAP2omega-green fluorescent protein (GFP) fusion protein targets vesicles to mitotic chromosomes in Xenopus A6 cells, suggesting that ZLAP2omega is involved in karyomere formation during early zebrafish development. When ZLAP2beta and gamma were expressed as GFP fusion proteins in Xenopus A6 cells, the beta- but not the gamma-isoform was found in association with mitotic chromosomes, and ZLAP2beta-containing chromosomes were decorated with vesicles. Further analysis of ZLAP2-GFP fusion proteins containing only distinct domains of the ZLAP2 isoforms revealed that the common N-terminal region in conjunction with beta- or omega-specific sequences mediate binding to mitotic chromosomes in vivo. PMID- 12734397 TI - Frizzled receptor dimerization is sufficient to activate the Wnt/beta-catenin pathway. AB - Wnt signaling has an important role in cell-fate determination, tissue patterning and tumorigenesis. Wnt proteins signal through seven-pass transmembrane receptors of the frizzled family to activate beta-catenin-dependent transcription of target genes. Using early Xenopus embryos, we show that frizzled receptors can dimerize and that dimerization is correlated with activation of the Wnt/beta-catenin pathway. Co-immunoprecipitation studies revealed that the receptor Xfz3 exists as a dimer when expressed in Xenopus embryos, and it has been shown to activate the Wnt/beta-catenin pathway as revealed by expression of the target gene siamois. Xfz3 dimerization requires intramolecular and/or intermolecular disulfide linkages, and the N-terminal extracellular region of the receptor, including the cysteine-rich domain (CRD), is sufficient for dimerization. The receptor Xfz7 behaves differently from Xfz3 when overexpressed in the embryo as Xfz7 is monomeric and is unable to directly activate the Wnt/beta-catenin pathway. However, activation of this pathway can be achieved by artificially forcing Xfz7 dimerization. These results provide the first direct evidence for the dimerization of frizzled receptors and suggest that dimerization contributes to transducing the Wnt/beta-catenin signal. PMID- 12734398 TI - An interaction between Sla1p and Sla2p plays a role in regulating actin dynamics and endocytosis in budding yeast. AB - The importance of a dynamic actin cytoskeleton for facilitating endocytosis has been recognised for many years in budding yeast and is increasingly recognised in mammalian cells. However, the mechanism for actin recruitment and the role it plays in endocytosis is unclear. Here we show the importance of two yeast proteins in this process. We demonstrate that Sla1p and Sla2p interact in vitro and in vivo and that this interaction is mediated by the central domain of Sla2p, which includes its coiled-coil region, and by a domain of Sla1p between residues 118 and 361. Overexpression of the interacting fragment of Sla1p causes reduced fluid-phase endocytosis and, interestingly, defects in subsequent trafficking to vacuoles. We show that Sla2p is required for the polarised localisation of Sla1p in cells but not for its cortical localisation or for its overlapping localisation with actin. Generation of an Deltasla1Deltasla2 double mutant demonstrates that Sla2p is likely to act upstream of Sla1p in endocytosis, whereas sensitivity to latrunculin-A suggests that the proteins have opposite effects on actin dynamics. We propose that Sla2p recruits Sla1p to endocytic sites. Sla1p and its associated protein Pan1p then regulate actin assembly through interactions with Arp2/3 and Arp2/3-activating proteins Abp1p and Las17/Bee1p. PMID- 12734399 TI - NF-kappaB signalling is inhibited by glucocorticoid receptor and STAT6 via distinct mechanisms. AB - NF-kappaB transcription factors are involved in the cellular response to stress, and are regulated by inhibitor (IkappaB) proteins, which prevent NF-kappaB mediated transcription by maintaining NF-kappaB in the cytoplasm. Proteins from other pathways are also known to regulate NF-kappaB negatively, notably the glucocorticoid receptor (GR) and IL-4-responsive STAT6. Both pathways were shown to inhibit NF-kappaB-mediated transcription, by expressing either STAT6 or GR and activating the respective pathways. Using fluorescent fusion proteins, we show that GR alters the timing of activated p65 NF-kappaB nuclear occupancy by increasing the export rate of p65 and is independent of whether GR is present as a dimer or monomer. Expression of STAT6 was also shown to alter p65 nuclear occupancy but appeared to affect the import rate and hence the overall maximal level of p65 translocation. Activating STAT6 with IL-4 prior to activating NF kappaB significantly increased this inhibition. Investigation of IkappaBa showed that activated STAT6 inhibited TNFalpha-mediated IkappaBa phosphorylation and degradation, whereas GR activation did not alter IkappaBalphakinetics. This demonstrates a clear separation of two distinct mechanisms of inhibition by STAT6 and GR upon the NF-kappaB pathway. PMID- 12734400 TI - IRSp53 is colocalised with WAVE2 at the tips of protruding lamellipodia and filopodia independently of Mena. AB - The insulin receptor tyrosine kinase substrate p53 (IRSp53) links Rac and WAVE2 and has been implicated in lamellipodia protrusion. Recently, however, IRSp53 has been reported to bind to both Cdc42 and Mena to induce filopodia. To shed independent light on IRSp53 function we determined the localisations and dynamics of IRSp53 and WAVE2 in B16 melanoma cells. In cells spread well on a laminin substrate, IRSp53 was localised by antibody labelling at the tips of both lamellipodia and filopodia. The same localisation was observed in living cells with IRSp53 tagged with enhanced green florescence protein (EGFP-IRSp53), but only during protrusion. From the transfection of deletion mutants the N-terminal region of IRSp53, which binds active Rac, was shown to be responsible for its localisation. Although IRSp53 has been reported to regulate filopodia formation with Mena, EGFP-IRSp53 showed the same localisation in MVD7 Ena/VASP (vasodilator stimulated phosphoprotein) family deficient cells. WAVE2 tagged with DsRed1 colocalised with EGFP-IRSp53 at the tips of protruding lamellipodia and filopodia and, in double-transfected cells, the IRSp53 signal in filopodia decreased before that of WAVE2 during retraction. These results suggest an alternative modulatory role for IRSp53 in the extension of both filopodia and lamellipodia, through WAVE2. PMID- 12734402 TI - The SH4-Unique-SH3-SH2 domains dictate specificity in signaling that differentiate c-Yes from c-Src. AB - c-Src and c-Yes are highly homologous members of the Src family of non-receptor tyrosine kinases. The overall sequence similarity between c-Src and c-Yes allows them to perform many overlapping functions. However, the phenotypes of the c-src and c-yes knockout mice, and cells derived from them, are quite different, indicating functional specificity between the two proteins. Specifically, c-src-/ cells are deficient in several processes that require dynamic regulation of the actin cytoskeleton. In order to begin to understand why c-Yes is unable to compensate for c-Src signaling, we used a series of Src/Yes chimeras in which the non-catalytic functional domains of Src527F were replaced by those of c-Yes. Using chicken embryo fibroblasts as a model system, our results indicate that the c-Yes N-terminal SH4-Unique domains are sufficient to inhibit the ability of Src527F to alter cell morphology, induce actin filament rearrangements or stimulate motility or invasive potential. The data also indicate that the SH4 Unique-SH3-SH2 domains of c-Yes work cooperatively and prevent activation of signaling proteins associated with Src527F transformation, including activation of phosphatidylinositol 3-kinase, phosphorylation of c-Raf and Akt and downregulation of RhoA-GTP. These data indicate that c-Yes may not modulate signals associated with c-Src-induced changes in actin filament integrity and may explain why c-Yes fails to compensate for c-Src signaling in src-/- cells. PMID- 12734401 TI - Contributions of molecular binding events and cellular compliance to the modulation of leukocyte adhesion. AB - The interaction of leukocyte function-associated antigen-1 (LFA-1) and intercellular adhesion molecule-1 (ICAM-1) is central to the regulation of adhesion in leukocytes. In this report, we investigated the mechanisms by which phorbol myristate acetate (PMA) promotes LFA-1-dependent cell adhesion. The adhesion of PMA-stimulated cells to immobilized ICAM-1 was quantified in direct force measurements acquired by atomic force microscopy (AFM). Enhanced adhesion of PMA-stimulated cells to immobilized ICAM-1 stemmed from an increase in the number of LFA-1-ICAM-1 complexes formed between the two apposing surfaces on contact, rather than by affinity modulation of LFA-1. Single molecule force measurements revealed that the force spectrum of the LFA-1-ICAM-1 complex formed by PMA-stimulated cells is identical to the force spectrum of the complex formed by resting cells. Thus, PMA stimulation does not modify the mechanical strength of the individual LFA-1-ICAM-1 interaction. Instead, the enhanced cell adhesion of PMA-stimulated cells appears to be a complex process that correlates with changes in the mechanical properties of the cell. We estimate that changes in the elasticity of the cell gave rise to a more than 10-fold increase in cell adhesion. PMID- 12734403 TI - Increased ploidy and KAR3 and SIR3 disruption alter the dynamics of meiotic chromosomes and telomeres. AB - We investigated the sequence of chromosomal events during meiotic prophase in haploid, diploid and autotetraploid SK1 strains of Saccharomyces cerevisiae. Using molecular cytology, we found that meiosis-specific nuclear topology (i.e. dissolution of centromere clustering, bouquet formation and meiotic divisions) are significantly delayed in polyploid SK1 meiosis. Thus, and in contrast to the situation in plants, an increase in ploidy extends prophase I in budding yeast. Moreover, we found that bouquet formation also occurs in haploid and diploid SK1 meiosis deficient in the telomeric heterochromatin protein Sir3p. Diploid sir3Delta SK1 meiosis showed pleiotropic defects such as delayed centromere cluster resolution in a proportion of cells and impeded downstream events (i.e. bouquet formation, homologue pairing and meiotic divisions). Meiotic telomere clustering occurred in diploid and haploid sir3Delta strains. Using the haploid system, we further show that a bouquet forms at the kar3Delta SPB. Comparison of the expression of meiosis-specific Ndj1p-HA and Zip1p in haploid control and kar3Delta time courses revealed that fewer cells enter the meiotic cycle in absence of Kar3p. Elevated frequencies of bouquets in kar3Delta haploid meiosis suggest a role for Kar3p in regulation of telomere dynamics. PMID- 12734405 TI - Vaccinia complement control protein: multi-functional protein and a potential wonder drug. AB - Vaccinia virus complement control protein (VCP) was one of the first viral molecules demonstrated to have a role in blocking complement and hence in the evasion of host defense. Structurally it is very similar to the human C4b-BP and the other members of complement control protein. Functionally it is most similar to the CR1 protein. VCP blocks both major pathways of complement activation. The crystal structure of VCP was determined a little over a year ago and it is the only known structure of an intact and complete complement control protein. In addition to binding complement, VCP also binds to heparin. These two binding abilities can take place simultaneously and contribute to its many function and to its potential use in several inflammatory diseases, e.g. Alzheimer's disease (AD), CNS injury, xenotransplantation, etc. making it a truly fascinating molecule and potential drug. PMID- 12734404 TI - Viral mimicry of the complement system. AB - The complement system is a potent innate immune mechanism consisting of cascades of proteins which are designed to fight against and annul intrusion of all the foreign pathogens. Although viruses are smaller in size and have relatively simple structure, they are not immune to complement attack. Thus, activation of the complement system can lead to neutralization of cell-free viruses, phagocytosis of C3b-coated viral particles, lysis of virus-infected cells, and generation of inflammatory and specific immune responses. However, to combat host responses and succeed as pathogens, viruses not only have developed/adopted mechanisms to control complement, but also have turned these interactions to their own advantage. Important examples include poxviruses, herpesviruses, retroviruses, paramyxoviruses and picornaviruses. In this review, we provide information on the various complement evasion strategies that viruses have developed to thwart the complement attack of the host. A special emphasis is given on the interactions between the viral proteins that are involved in molecular mimicry and the complement system. PMID- 12734406 TI - Lessons in detente or know thy host: the immunomodulatory gene products of myxoma virus. AB - The poxvirus, myxoma virus, encodes within its genome at least eleven different proteins that compromise, skew, or disable the innate and adaptive responses of its hosts. In the laboratory rabbit, Oryctolagus cuniculus, these effects result in myxomatosis, a fatal condition characterized by skin lesions and systemic immunosuppression. Interestingly, while myxoma infection also causes skin lesions in its natural host and in natural populations of O. cuniculus in Australia where this novel host and the virus have co-evolved, the condition of myxomatosis does not ensue and infection is not fatal. In this review I discuss the biochemical properties of the characterized immunomodulatory proteins of myxoma virus, and their pathogenic effects in laboratory rabbits. Disruption of any one myxoma immunomodulatory gene diminishes the severity of the infection without compromising infectivity. Thus, the characterized immunomodulatory genes appear not to be required for a productive infection in vivo. The differences in the severity of their effects in laboratory-bred versus wild O. cuniculus suggest that the outcome of myxoma infection is a consequence of the interplay between the viral immunomodulatory gene products and the cells and molecules of the host immune system. PMID- 12734408 TI - Variability or conservation of hepatitis C virus hypervariable region 1? Implications for immune responses. AB - The hypervariable region 1 (HVR1) of the E2 protein of hepatitis C virus (HCV) is highly heterogeneous in its primary sequence and is responsible for significant inter- and intra-individual variation of the infecting virus, which may represent an important pathogenetic mechanism leading to immune escape and persistent infection. A binding site for neutralizing antibodies (Ab) has also been allegedly identified in this region. Prospective studies of serological responses to synthetic oligopeptides derived from naturally-occurring HVR1 sequences showed promiscuous recognition of HVR1 variants in most patients via binding to C terminal amino acid residues with conserved physicochemical properties. Monoclonal antibodies generated by immunization of mice with peptides derived from natural HVR1 sequences were shown to recognize several HVR1 variants in line with evidence gathered from studies using human sera. In addition, selected mAbs were able to bind HVR1 in the context of a complete soluble form of the E2 glycoprotein, indicating recognition of correctly folded sequences, and were shown to specifically capture circulating and recombinant HCV particles, suggesting that HVR1 is expressed on intact virus particles and therefore potentially able to interact with cellular receptor(s). These findings suggest that it is possible to induce a broadly reactive clonal immune response to multiple HCV variants and that this mechanism could be used in principle to induce protective immunity for a large repertoire of HCV variants. PMID- 12734407 TI - The hepatitis C virus persistence: how to evade the immune system? AB - Hepatitis C virus (HCV) is an emerging virus of medical importance. A majority of HCV infections become chronic and lead to chronic hepatitis, liver cirrhosis and hepatocellular carcinoma. HCV usually induces robust immune responses, but it frequently escapes the immune defense to establish persistent infection. The fact that HCV exists as an evolving quasispecies plays an important role in the selection of escape mutants. Furthermore, several viral proteins interfere with cellular functions, in particular, those involved in the immune response of the host. Several HCV proteins also modulate cell signalling through interaction with different effectors involved in cell proliferation and apoptosis, or in the interferon-signalling pathway. In addition, HCV infects immune cells such as B and T cells, and thus affects their normal functions. These various strategies used by HCV to counter the immune response of the host are reviewed here. A better understanding of these mechanisms would help design new therapeutic targets. PMID- 12734409 TI - Regulatory mechanisms of viral hepatitis B and C. AB - Of all the hepatitis viruses, only the hepatitis B virus (HBV) and hepatitis C virus (HCV) cause chronic hepatitis, which can progress to cirrhosis and hepatocellular carcinoma. In this review, we discuss how these two biologically diverse viruses use common pathways to induce oxidative stress and activation of key transcription factors, known to be involved in inflammatory processes in cells. Activation of NF-kB and STAT-3 most likely contribute to the progression of viral infections to chronic hepatitis and liver oncogenesis associated with HBV and HCV infections. In this review, we focus on the mechanisms of action of HBx and HCV NS5A proteins in inducing intracellular events associated with the viral infections. PMID- 12734410 TI - HIV-1 Nef control of cell signalling molecules: multiple strategies to promote virus replication. AB - HIV-1 has at its disposal numerous proteins encoded by its genome which provide the required arsenal to establish and maintain infection in its host for a considerable number of years. One of the most important and enigmatic of these proteins is Nef. The Nef protein of HIV-1 plays a fundamental role in the virus life cycle. This small protein of approximately 27 kDa is required for maximal virus replication and disease progression. The mechanisms by which it is able to act as a positive factor during virus replication is an area of intense research and although some controversy surrounds Nef much has been gauged as to how it functions. Its ability to modulate the expression of key cellular receptors important for cell activation and control signal transduction elements and events by interacting with numerous cellular kinases and signalling molecules, including members of the Src family kinases, leading to an effect on host cell function is likely to explain at least in part its role during infection and represents a finely tuned mechanism where this protein assists HIV-1 to control its host. PMID- 12734411 TI - Molecular interactions of 'high risk' human papillomaviruses E6 and E7 oncoproteins: implications for tumour progression. AB - The aetiology of cervical cancer has been primarily attributed to human papillomaviruses (HPVs). These are characterized by the persistent expression of the two oncogenes, E6 and E7. Experimental studies show that E6 and E7 genes of the high risk HPVs deregulate key cell cycle controls. Recent work has uncovered new cellular partners for these proteins that throw light on many of the pathways and processes in which these viral proteins intervene. This review focuses on the regulation of host proteins by the viral oncoproteins and consequence of such interactions on cell survival, proliferation, differentiation and apoptosis. PMID- 12734413 TI - Hsp90 and chromatin: where is the link? PMID- 12734415 TI - DNA building blocks at the foundation of better survival. PMID- 12734412 TI - Host-pathogen interactions during apoptosis. AB - Host pathogen interaction results in a variety of responses, which include phagocytosis of the pathogen, release of cytokines, secretion of toxins, as well as production of reactive oxygen species (ROS). Recent studies have shown that many pathogens exert control on the processes that regulate apoptosis in the host. The induction of apoptosis upon infection results from a complex interaction of parasite proteins with cellular host proteins. Abrogation of host cell apoptosis is often beneficial for the pathogen and results in a successful host invasion. However, in some cases, it has been shown that induction of apoptosis in the infected cells significantly imparts protection to the host from the pathogen. There is a strong correlation between apoptosis and the host protein translation machinery: the pathogen makes all possible efforts to modify this process so as to inhibit cell suicide and ensure that it can survive and, in some cases, establish latent infection. This review discusses the significance of various pathways/steps during virus-mediated modulation of host cell apoptosis. PMID- 12734416 TI - The spindle-associated transmembrane protein Axs identifies a new family of transmembrane proteins in eukaryotes. AB - Axs mutations disrupt both the progression of the meiotic cell cycle and meiotic chromosome segregation in Drosophila. Axs protein co-localizes with endoplasmic reticulum components and is present within a novel structure ensheathing the meiotic spindle. We show that Axs encodes the founding member of a eukaryotic family of trans-membrane proteins. PMID- 12734414 TI - PARC: a potential target for cancer therapy. PMID- 12734417 TI - Altered mRNA translation: possible mechanism for CML disease progression. PMID- 12734418 TI - Direct regulation of RNA polymerase III transcription by RB, p53 and c-Myc. AB - The synthesis of tRNA and 5S rRNA by RNA polymerase (pol) III is cell cycle regulated in higher organisms. Overexpression of pol III products is a general feature of transformed cells. These observations may be explained by the fact that a pol III-specific transcription factor, TFIIIB, is strongly regulated by the tumor suppressors RB and p53, as well as the proto-oncogene product c-Myc. RB and p53 repress TFIIIB, but this restraint can be lost in tumors through a variety of mechanisms. In contrast, c-Myc binds and activates TFIIIB, causing potent induction of pol III transcription. Using chromatin immunoprecipitation and RNA interference, we show that c-Myc interacts with tRNA and 5S rRNA genes in transformed cervical cells, stimulating their expression. Availability of pol III products may be an important determinant of a cell's capacity to grow. The ability to regulate pol III output may therefore be integral to the growth control functions of RB, p53 and c-Myc. PMID- 12734419 TI - Coordination of S-phase events and genome stability. PMID- 12734420 TI - The ATR-independent DNA replication checkpoint. PMID- 12734421 TI - Gleevec resistance: lessons for target-directed drug development. PMID- 12734422 TI - A novel role for the hypoxia inducible transcription factor HIF-1alpha: critical regulation of inflammatory cell function. PMID- 12734423 TI - Clinical impact of ATR checkpoint signalling failure in humans. PMID- 12734424 TI - Hedgehog signaling: progenitor phenotype in small-cell lung cancer. AB - Recently, we have shown that small cell lung cancer (SCLC) is dependent on activation of Hedgehog signaling, an embryonic pathway implicated in development, morphogenesis and the regulation of stem cell fates. These findings form the framework for an emerging view of cancer as a process of aberrant organogenesis in which progenitor/ stem cells escape dependence on niche signaling through mutation in genes such as Ptch, or through persistent activation of progenitor cell pathways. Interestingly, the normally quiescent airway epithelial compartment uses the Hh pathway to repopulate itself when challenged by injury. How Hh signaling works to promote the malignant phenotype promises to be as important biologically as the promise of Hh pathway inhibitors are clinically. PMID- 12734425 TI - Role of JNK in tumor development. AB - The c-Jun NH2-terminal kinase (JNK) is implicated in oncogenic transformation. However, studies of the effect of Jnk gene disruption on Ras-induced transformation of murine fibroblasts indicate that JNK may act as a suppressor of Ras transformation and that the JNK signaling pathway contributes to the apoptotic elimination of transformed cells in vivo. The conclusion that JNK can act as a tumor suppressor is consistent with the presence of loss-of-function mutations in JNK pathway components (Jnk3 and Mkk4) in human tumors. Nevertheless, JNK can also contribute to the proliferation and survival responses of some tumors. A key question that remains unresolved concerns the genetic and mechanistic basis for these different roles of JNK in tumors. Indeed, an understanding of this question will be required for the rational use of small molecule inhibitors of JNK for tumor therapy. PMID- 12734426 TI - Multistep carcinogenesis: a chain reaction of aneuploidizations. AB - Carcinogenesis is a multistep process in which new, parasitic and polymorphic cancer cells evolve from a single, normal diploid cell. This normal cell is converted to a prospective cancer cell, alias "initiated", either by a carcinogen or spontaneously. The initiated cell typically does not have a new distinctive phenotype yet, but evolves spontaneously--over months to decades--to a clinical cancer. The cells of a primary cancer also evolve spontaneously towards more and more malignant phenotypes. The outstanding genotype of initiated and cancer cells is aneuploidy, an abnormal balance of chromosomes, which increases and varies in proportion with malignancy. The driving force of the spontaneous evolution of initiated and cancerous cells to ever more abnormal phenotypes is said to be their "genetic instability". However, since neither the instability of cancer phenotypes nor the characteristically slow kinetics of carcinogenesis are compatible with gene mutation, we propose here that the driving force of carcinogenesis is the inherent instability of aneuploid karyotypes. Aneuploidy renders chromosome structure and segregation error-prone, because it unbalances mitosis proteins and the many teams of enzymes that synthesize and maintain chromosomes. Thus, carcinogenesis is initiated by a random aneuploidy, which is induced either by a carcinogen or spontaneously. The resulting karyotype instability sets off a chain reaction of aneuploidizations, which generate ever more abnormal and eventually cancer-specific combinations and rearrangements of chromosomes. According to this hypothesis the many abnormal phenotypes of cancer are generated by abnormal dosages of thousands of aneuploid, but un-mutated genes. PMID- 12734427 TI - Break the cycle: the role of cell-cycle modulation in the prevention of vasculoproliferative diseases. AB - Atherosclerosis is a major health problem in the western world, and the effectiveness of interventional therapeutic modalities for symptomatic atherosclerotic lesions is limited by vessel restenosis. Proliferation and migration of smooth muscle cells (SMCs) play a central role in post interventional restenosis. Accordingly, many therapeutic approaches attempt to inhibit SMC proliferation. As the cell cycle is a final common pathway in SMC proliferation, proteins of the cell cycle have emerged as logical targets for the treatment and prevention of restenotic lesions. In this review we discuss current approaches that target the cell cycle in smooth muscle cells, and also describe recent and ongoing clinical trials that involve cell-cycle manipulation in the treatment of vasculoproliferative diseases. PMID- 12734428 TI - Akt and PI 3-kinase signaling in cardiomyocyte hypertrophy and survival. AB - In many systems, activation of the "protein and lipid kinase" phosphoinositide 3 kinase (PI 3-kinase) and its downstream serine-threonine kinase effector, Akt (or Protein Kinase B), provide a potent stimulus for cell proliferation, growth, and survival. In the heart, constrained by the limited proliferative capacity of cardiomyocytes, this pathway plays a key role in regulating cardiomyocyte growth and survival, with little effect on proliferation. Simultaneously, PI 3-kinase and Akt are important modulators of metabolic substrate utilization and cardiomyocyte function. Thus, the convergent signaling pathways controlling so many clinically important phenotypes of the cardiomyocyte suggest it holds promise as a therapeutic target in a variety of cardiac diseases. However, the similar role of PI 3-kinase/Akt signaling in neoplasia suggests the difficulty of activating this pathway in the heart without invoking adverse consequences elsewhere. Here we review evidence regarding the role of PI 3-kinase/Akt in controlling cardiomyocyte growth and survival, and discuss the implications for therapeutic strategies. PMID- 12734429 TI - Genetic instability, DNA damage and atherosclerosis. AB - We review the role of somatic mutations and genetic instability in the pathogenesis of atherosclerosis, suggesting novel therapeutic approaches. PMID- 12734430 TI - Fine tuning of kinetochore function by phosphorylation. PMID- 12734431 TI - Replication-coupled topoisomerase II templates the mitotic chromosome scaffold? PMID- 12734432 TI - p53 in a crosstalk between DNA repair and cell cycle checkpoints. PMID- 12734433 TI - Loss of mitotic spindle checkpoint activity predisposes to chromosomal instability at early stages of fibrosarcoma development. AB - Mice carrying the bovine papillomavirus type I genome develop dermal fibrosarcomas in a multiple step process characterized by distinctive proliferative stages. Chromosomal aberrations are identified early in this tumorigenic pathway, however, the mechanism that originates them is unknown. Using a functional assay, we investigated the status of the mitotic spindle cell cycle checkpoint (MSCCC) that regulates the metaphase to anaphase transition, in cells representing different stages of fibrosarcoma progression. Loss of MSCCC activity was apparent in mild fibromatosis and completely abolished in aggressive fibromatosis and fibrosarcoma lesions. This altered MSCCC status was confirmed biochemically by deregulated expression of Cks1 protein and unscheduled cyclin B metabolism. Immunoprecipitation and sequencing analyses indicated that mutation of p53 was not required for the abrogation of the MSCCC. These results demonstrate that loss of mitotic spindle checkpoint activity predisposes to chromosomal instability at early stages of fibrosarcoma development. To our knowledge, these studies constitute the first report of a transition in MSCCC activity in a tumorigenesis model. PMID- 12734434 TI - Flavopiridol induces mitochondrial-mediated apoptosis in murine glioma GL261 cells via release of cytochrome c and apoptosis inducing factor. AB - Glioblastoma (GBM) remains one of the most challenging solid cancers to treat due to its highly proliferative, angiogenic and invasive nature. The small molecule CDK inhibitor, flavopiridol, has demonstrated antitumor activity in human xenograft models and is currently in clinical trials showing efficacy in patients with advanced disease. We have developed an experimental animal model using the murine glioma GL261 cells as a novel in vivo system to screen potential therapeutic agents for GBM. Results of in vitro testing demonstrate that flavopiridol has several relevant clinical characteristics such as its ability to: 1. inhibit cell growth; 2. inhibit cell migration; 3. decrease expression of cyclin D1, CDK4 and p21; 4. induce apoptosis in cells with high levels of p27 expression; and 5. decrease the expression of the anti-apoptotic protein Bcl-2. The mechanism by which flavopiridol induces apoptosis is mitochondrial-mediated. We demonstrate by electron microscopy and immunohistochemistry that drug treatment induces mitochondrial damage that was accompanied by the release of cytochrome c into the cytosol together with the translocation of apoptosis inducing factor (AIF) into the nucleus. This finding in murine glioma cells differs from the mechanism of flavopiridolinduced cell death reported by us for human glioma cells (Alonso et al., Mol Cancer Ther 2003; 2:139) where drug treatment induced a caspase- and cytochrome c-independent pathway in the absence of detectable damage to mitochondria. In apoptotic human glioma cells only translocation of AIF into the nucleus occurred. Thus, the same drug kills different types of glioma cells by different mitochondrial-dependent pathways. PMID- 12734435 TI - Modulation of gene expression by lentiviral-mediated delivery of small interfering RNA. AB - RNA interference (RNAi) mediates sequence-specific posttranscriptional gene silencing triggered by double-stranded RNA. RNAi is an effective tool for functional genomics in C. elegans, whereas biochemical characteristics of RNAi hamper its use in mammalian cells. We here analyze kinetic and quantitative aspects of mammalian RNAi in different cell lines using novel lentiviral constructs with double H1-shRNA expression cassettes located in the U3 region of the LTR. Using enhanced green fluorescence protein (EGFP) as a target gene for RNAi and red fluorescence protein (RFP) as surrogate marker for intracellular siRNA expression, we show that long-term siRNA expression mediates stable RNAi. Furthermore, RNAi-induced gene silencing varies from minimal to complete loss-of function phenotypes within homogeneous and between different cell populations. Interestingly, the extent of gene silencing correlates to lentiviral integrations as well as siRNA expression levels in target cells. Finally, we demonstrate functional gene silencing of a cytokine receptor gene in normal CD34+ hematopoietic progenitor cells. These data suggest that the use of suitable lentiviral constructs to quantify siRNA expression in living cells may allow analysis of loss-of-function phenotypes in a dose-dependent manner in a wide variety of target cells. PMID- 12734436 TI - Pim-1 phosphorylates the DNA binding domain of c-Myb. AB - The c-Myb transcription factor regulates cellular differentiation and proliferation and is regulated by complex mechanisms that control its repressed oncogenic activity. The transcriptional activity of c-Myb is regulated by the serine/threonine protein kinase Pim-1. Here, we show that Pim-1 is able to interact with c-Myb and the closely related transcription factor A-Myb, via direct interactions with the highly conserved Myb DNA binding domain. Pim-1 associated with Myb both in cells and in vitro, and phosphorylated the Myb DNA binding domain, suggesting that it regulates Myb protein activity by direct phosphorylation. PMID- 12734437 TI - Pyoderma and scabies: a benign association? PMID- 12734438 TI - Staphylococcal epidermolysins. AB - PURPOSE OF REVIEW: Staphylococcal epidermolysins are the major causative toxins of bullous impetigo and staphylococcal scalded skin syndrome. This disease is characterized by the splitting of the epidermis between two cell layers resulting in exfoliation. It predominantly affects newborn babies and exposes them to secondary infections. This leads to the risk of epidemics, especially in nurseries. With only an experimental model which consists of skin injections in newborn mice and the recent determination of three-dimensional structures, the essential function of these toxins remained controversial, split between that of specific proteases and that of superantigens. RECENT FINDINGS: Staphylococcal epidermolysins now constitute a family of toxins, with the recent characterizations of two new serotypes: ETC and ETD. They may be secreted by sensitive or methicillin-resistant strains. Four molecules were also identified in Staphylococcus hyicus responsible for exudative epidermitis in swine. While different observations suggested a proteolytic action to these toxins, the histological parallel made with pemphigus foliaceus greatly helped in the characterization of the targets for epidermolysins ETA, ETB, ETD: desmoglein-1, a desmosome-constitutive protein, and incidentally melanocyte-stimulating hormones, which accounts for the blisters observed clinically. SUMMARY: The growing complexity in staphylococcal toxins has to be taken into account both for their association with diseases and for diagnosis purposes. Even though cases of staphylococcal scalded skin syndrome in adults are rare, they raise further questions about the pathogenic features of the disease such as individual sensitivity and distribution of the toxins into the body. PMID- 12734439 TI - Molecular diagnosis of nontuberculous mycobacteria. AB - PURPOSE OF REVIEW: Diagnosis of infection due to nontuberculous mycobacteria is not easy, as it must be distinguished from colonization or contamination by other nontuberculous mycobacteria. Molecular methods offer many advantages over conventional methods of identification. The results are obtained rapidly, are reliable and reproducible, and even mixed or contaminated cultures can be examined. This review highlights the recent advances in molecular techniques for identification of nontuberculous mycobacteria. RECENT FINDINGS: Nontuberculous mycobacteria are ubiquitous towards the environment and have the potential to colonize and cause serious infection. An increasing number of species and clinical presentations are being described, and progress has been made towards the understanding of the underlying predisposing factors. Disease caused by nontuberculous mycobacteria is often associated with various forms of immunosuppression, particularly HIV infection, whereas mild forms of immune defects have been observed in some patients who, apart from their nontuberculous mycobacterial disease, seem to be healthy on initial examination. Molecular techniques have shown their usefulness for the identification of most mycobacteria. Probes are widely used in clinical laboratories for the identification of the most common mycobacterial species. Because automated DNA sequencing and the programs for analysing sequence data have become technically simpler, polymerase chain reaction-based sequencing is now used in many mycobacterial reference laboratories as a routine method for species identification. SUMMARY: Significant advances have been made with molecular tools for diagnosis of mycobacteria. The DNA microarray technique holds great promise for the future because it is easy to perform, it can be readily automated, and it allows the identification of a large number of mycobacterial species in one reaction. PMID- 12734440 TI - Imiquimod. AB - PURPOSE OF REVIEW: Imiquimod is the first member of a new class of immune response modifiers; it was first approved in 1997 for the topical treatment of external genital and perianal warts. It is an imidazoquinoline, a novel synthetic compound which is an immune response stimulator, enhancing both the innate and acquired immune pathways (particularly T helper cell type 1-mediated immune responses) resulting in antiviral, antitumour and immunoregulatory activities. The mechanism of action of imiquimod involves cytokine induction in the skin, which then triggers the host's immune system to recognize the presence of a viral infection or tumour, ultimately to eradicate the associated lesion. RECENT FINDINGS: Imiquimod, a patient-applied topical 5% cream is clinically efficacious and safe in the management of condylomata acuminata and other warty manifestations of human papillomavirus infections. Although not licensed for use against other viral skin infections, preliminary data suggest imiquimod's success against molluscum contagiosum, caused by a poxvirus. Initial studies with imiquimod for the management of HPV-related intraepithelial dysplasias (bowenoid papulosis/vulvar intraepithelial neoplasia) as well as for ultraviolet-induced skin lesions such as actinic keratoses, Bowen's disease, and basal cell carcinomas show great promise in immunocompetent and immunosuppressed patients. SUMMARY: In the future, imiquimod and newer generations of imidazoquinolines (resiquimod) require further investigation for potential clinical utility in treating other cutaneous and mucosal viral infections, dysplasias and neoplasia, as well as potential vaccine adjuvants. PMID- 12734441 TI - Melanin and fungi. AB - PURPOSE OF REVIEW: Production of melanin has been associated with virulence in diverse microorganisms. Melanization of fungi has been noted for many years in predominantly subcutaneous infections such as chromoblastomycosis and more recently most extensively studied in a yeast causing systemic infection, Cryptococcus neoformans. Pigmented fungi are increasingly important human pathogens and currently available antifungals are often sub-optimal for serious infections. This review focuses on recent publications on melanin in fungi with particular reference to the role of melanin in virulence, protection against antifungal drugs, and promoting survival in the environment. RECENT FINDINGS: Inhibition of melanin production by C. neoformans can prolong survival of lethally infected mice. In contrast, melanin in C. neoformans and Histoplasma capsulatum yeast cells can bind amphotericin B and caspofungin, thereby reducing the fungicidal affects of these drugs. H. capsulatum and Paracoccidioides brasiliensis have only recently been shown to produce melanin in vitro and during infection. Additionally, melanin derived from melanized C. neoformans yeast and Aspergillus niger conidia can activate complement, which may modify immune responses to infection. Studies on C. neoformans laccase have revealed that the enzyme is located on the cell wall, which may allow for interactions with the host. Melanization reduces the susceptibility of C. neoformans to enzymatic degradation and toxicity from a heavy metal, which may afford protection to the fungus against similar insults in the environment. SUMMARY: Melanin has been referred to as 'fungal armor' due to the ability of the polymer to protect microorganisms against a broad range of toxic insults. Recent publications continue to reveal important contributions of melanin to survival of fungi in the environment and during infection. PMID- 12734442 TI - Diagnosis of cutaneous leishmaniasis. AB - PURPOSE OF REVIEW: The purpose of this review is to discuss some of the most important issues in the clinical and laboratory diagnosis of cutaneous leishmaniasis. The former is based on clinical experience and the latter on the literature published within the last couple of years. The sensitivity and specificity of old and novel tests to identify the parasite, antigenic products, the host's immune response, and the molecular diagnosis by the PCR are considered here. RECENT FINDINGS: Recent clinical and laboratory research has addressed the individual, entomological, and epidemiological aspects of acute and chronic forms of cutaneous leishmaniasis. Traditional diagnostic investigations for individual cases include the search of suggestive history and clinical features, the identification of amastigotes by histology or direct microscopy, and the growth of promastigotes in culture. Diagnosis by the polymerase chain reaction seems to be approaching a 'gold standard' status as novel techniques offer considerable advantages in the collection and transport of specimens and DNA extraction procedures that are more efficient in individual and field-based protocols. Several authors have reported consistent 100% specificity with increasing sensitivity that overall is between 92 and 98%. Moreover the PCR approach makes possible the fast identification at the Leishmania species and subspecies level. SUMMARY: The diagnostic tools discussed in this review are relevant not only for the diagnosis in endemic regions of the world but also for travellers and workers that following exposure return to non-endemic countries. PMID- 12734443 TI - Staphylococcal resistance revisited: community-acquired methicillin resistant Staphylococcus aureus--an emerging problem for the management of skin and soft tissue infections. AB - PURPOSE OF REVIEW: In the community non-localized or deep staphylococcal skin and soft tissue infections are typically managed with beta-lactamase stable penicillins. The aims of this review are (1) to evaluate the evidence for the emergence of new strains of community-acquired methicillin resistant Staphylococcus aureus (MRSA), (2) to identify the reasons for their significant association with cutaneous infections, and (3) to consider how they arose and how big a threat they pose to the management of such infections outside hospitals. RECENT FINDINGS: MRSA are emerging as significant community pathogens, especially in previously healthy children with no recognizable risk factors, and are predominantly associated with skin and soft tissue infections (especially abscesses and cellulitis). When present, risk factors are generally similar to those for infection with methicillin susceptible S. aureus. The MRSA isolates associated with such infections may not be entirely 'new', but could represent the displacement of some hospital clones (e.g. EMRSA-15 or variants thereof) to the community as well as the de-novo generation of novel MRSA clones by multiple horizontal transmissions of the mecA gene into methicillin susceptible S. aureus with different genetic backgrounds, some of which are already circulating globally. Community-acquired MRSA from diverse locations are non multiresistant and almost always contain the novel type IV SCCmec commonly found in coagulase negative staphylococci, but also in hospital-associated gentamicin susceptible MRSA from France, the paediatric clone and in EMRSA-15. SUMMARY: More local data on CA-MRSA infections are needed so that dermatologists and community physicians can assess the risk of such infections amongst their patients and avoid the inappropriate administration of beta-lactams. No simple change in prescribing practices will entirely alleviate selective pressure for the spread of community acquired MRSA and not exacerbate resistance in pyogenic streptococci, commonly found together with S. aureus in skin and soft tissue infections. The importance of hygiene in preventing the spread of community-acquired MRSA in the community must be reemphasized. PMID- 12734444 TI - Viral respiratory tract infections: increasing importance and a new pathogen. PMID- 12734445 TI - Respiratory infections caused by non-typeable Haemophilus influenzae. AB - PURPOSE OF REVIEW: This review will consider recent developments in the clinical aspects of infections due to non-typeable Haemophilus influenzae. In addition, newer developments in the areas of mechanisms of pathogenesis, host pathogen interaction, immune responses and efforts toward vaccine development will be reviewed briefly. RECENT FINDINGS: Non-typeable H. influenzae continues to be a common cause of otitis media in infants and children, sinusitis in children and adults, pneumonia in adults, and lower respiratory tract infection in adults with chronic obstructive pulmonary disease. While the rate of beta-lactamase production by isolates of H. influenzae varies geographically, most regions show a rate of 20-35% of isolates producing beta-lactamase. Recent studies have highlighted the possible role of bacterial biofilms formed by H. influenzae as a cause of otitis media. Several lines of evidence indicate that H. influenzae causes intracellular infection in the lower respiratory tract in chronic obstructive pulmonary disease and this observation has important implications in understanding the human immune response to the bacterium. Lipooligosaccharide is an important virulence factor for H. influenzae and research is generating new information on the complex role of this molecule in colonization and infection of the respiratory tract. Several surface molecules are under active evaluation as vaccine antigens. SUMMARY: Non-typeable H. influenzae is an important cause of respiratory tract infections in children and adults. Most strains are susceptible to amoxicillin/clavulanate, fluoroquinolones and the newer macrolides. Research in the next decade promises substantial progress in the challenge of developing vaccines for nontypeable H. influenzae. PMID- 12734446 TI - Pseudomonas aeruginosa pneumonia. AB - PURPOSE OF REVIEW: Recent articles of clinical interest on Pseudomonas aeruginosa respiratory tract infections including CAP, nosocomially-acquired pneumonia, particularly in the ventilated patient, and chronic infections in cystic fibrosis patients are reviewed. RECENT FINDINGS: The growing importance of P. aeruginosa as an etiologic agent of CAP, the occurrence of CAP in previously healthy adults and its high prevalence as an etiologic agent of late VAP are stressed in recent studies. The effect of antibiotics on the recovery of bacteria in respiratory samples of patients with VAP can be marked and as early as 12 h after administration of antimicrobials certain organisms are no longer cultivable; in contrast, P. aeruginosa can still be recovered even after 48 h of adequate therapy. Type III secretory proteins are recognized as important virulent factors in P. aeruginosa. This phenotype predicts a worse outcome in patients with VAP. Fluoroquinolones have a major role in the emergence of multiply resistant P. aeruginosa in patients with VAP. Pharmacokinetic/pharmacodynamic parameters of antimicrobials with antipseudomonal activity are gaining importance as a means of optimization of antibiotic therapy. In CF, the knowledge of the pharmacokinetics and bioavailability of inhaled tobramycin and its long term beneficial effect in lung function are important developments in this area. SUMMARY: P. aeruginosa continues to be a serious problem worldwide as a cause of respiratory tract infections in selected populations. Microbiologic diagnosis remains difficult and plagued with pitfalls. The application of modern PK/PD concepts should help to optimize antibiotic therapy of this increasingly difficult to treat infection, particularly at the respiratory tract level and with an increasing prevalence of resistance to all antipseudomonal agents. Inhaled antibiotics, particularly tobramycin, are increasingly used for the prevention and treatment of P. aeruginosa infection in CF patients. PMID- 12734447 TI - Legionella spp.: community acquired and nosocomial infections. AB - PURPOSE OF REVIEW: The key points of this review are the increasingly recognized risk of home-acquired Legionnaires' disease; the significance and potential pathogenic role of other species of Legionella spp., different from L. pneumophila, and of other microorganisms that are phylogenetically close to Legionella and that have been named as Legionella-like amoebal pathogens; the breakthrough in the diagnosis of the disease caused by new commercially available urine antigen detection tests; the controversy over sensitivity and specificity of serological diagnostic methods; the recognition of a variety of possible mixed infections, particularly in the immunocompromised population; and new and controversial aspects of the therapeutic approach to legionellosis. RECENT FINDINGS: During the last year a number of articles have provided clinically relevant insights into our knowledge of Legionnaires' disease. In view of the fact that Legionella spp. have progressively become recognized as relatively common causative agents of both community-acquired and nosocomial legionellosis, this is an opportune moment for this review. SUMMARY: If domestic aquatic reservoirs were eventually confirmed as significant agents of transmission of legionellosis, the adoption of preventive measures would then be crucial. The progressive identification of other species, different from L. pneumophila, as causative agents of pneumonia should both encourage microbiologists and clinicians to improve their diagnostic methodology and increase the awareness of these infections. Finally, the awareness of mixed infections, probably far more severe and perhaps not so uncommon as previously thought, has important clinical connotations for both the diagnostic and the therapeutic approach to legionellosis in the immunosuppressed host, particularly in those cases of delayed clinical resolution. PMID- 12734448 TI - Severe community-acquired pneumonia: what's in a name? AB - PURPOSE OF REVIEW: Formerly, patients with community-acquired pneumonia admitted to an intensive care unit were considered as having the severe form of the disease. Recently, guidelines have distinguished severe and non-severe community acquired pneumonia based on clinical definitions. In this review, we describe the different definitions of severe community-acquired pneumonia, and whether a differentiation based on these definitions reflects variation in etiology, risk factors, diagnostic approaches and treatment. RECENT FINDINGS: New definitions do not seem to accurately identify patients with high risks of mortality; patients not admitted to an intensive care unit could also be diagnosed as having severe community-acquired pneumonia. Host-factors, such as genetic factors and underlying diseases, can influence severity of presentation of community-acquired pneumonia. Distribution of pathogens in severe and non-severe disease forms is comparable. Initial antibiotic therapy in patients with severe disease should provide coverage of Streptococcus pneumoniae and Legionella pneumophila, as delay is associated with worse outcomes. However, recent studies also suggested an additional benefit of atypical coverage in non-severe disease. As a result, initial therapy with a beta-lactam plus a macrolide or an anti-pneumococcal fluoroquinolone is recommended for all patients with community-acquired pneumonia. Furthermore, the value of vaccination against pneumococci to prevent episodes of severe disease is yet unknown. SUMMARY: As current guidelines do not adequately identify patients with high risk of mortality and intensive care unit admittance, clinical judgment remains important. Based on distribution of pathogens, investigational procedures and therapy recommended in recent guidelines, differentiation between severe and non-severe community-acquired pneumonia does not seem useful. Whether atypical coverage indeed has additional value in non-severe or pneumococcal CAP, however, remains to be determined. In addition, the preventive benefit of influenza and pneumococcal vaccination for development of SCAP awaits further evidence. PMID- 12734449 TI - Time for non-cancer. PMID- 12734450 TI - Administration of as required subcutaneous medications by lay carers: developing a procedure and leaflet. AB - Good symptom control is considered one of the factors that determines whether patients are able to die at home. This article describes a practice development project to enable carers to administer as required (PRN) subcutaneous medication, via a subcutaneous butterfly device in a community setting. Medications were mainly analgesic injections, although others, such as midazolam, were also given. The article highlights the lack of literature on the topic, reviews relevant background information and presents two successful case histories. It outlines the development of a procedure and accompanying leaflet to support community nurses in teaching relatives how to carry out this task, and the implementation of the process. PMID- 12734451 TI - Non-pharmacological management of breathlessness: a collaborative nurse- physiotherapist approach. AB - Breathlessness is a common and challenging symptom in palliative care. A simple audit of 470 patients under the St Christopher's Hospice Homecare team found that 43% complained of breathlessness. Research has indicated that non-pharmacological management can be of benefit to terminally ill patients with breathlessness. Knowing that both nurses and physiotherapists each have core skills to offer these patients, the authors aimed to integrate and consolidate their approach to enhance best practice. The supportive care and advice of the clinical nurse specialist was combined with physiotherapy sessions. The aim of this collaborative approach was to reduce patient anxiety, maximize respiratory function and provide information to help patients understand, adapt and regain some control of their breathing. A leaflet containing simple written information for patients was developed and found to be useful. It was found that supportive professional relationships strengthened links between the multiprofessional team. Positive feedback has been received from patients and health-care professionals about the sessions and breathlessness leaflet. By establishing this combined approach the authors have contributed to significant practice development at St Christopher's Hospice. PMID- 12734452 TI - Nurses and health support workers' views on cardiopulmonary resuscitation in a hospice setting. AB - This paper reports on a study which took place in a 15-bed hospice and provides an account of the views of registered nurses (n = 12) and health support workers (n = 10) in relation to cardiopulmonary resuscitation (CPR) within a hospice setting. The study was part of a larger ethnographic research project being conducted in the hospice when the British Medical Association/Royal College of Nursing and the Resuscitation Council UK (Resuscitation Council UK, 2001) published guidelines urging all establishments that face decisions about attempting CPR to develop local policies. The guidelines provided the impetus for the present study, the aim of which was to explore staff responses to the introduction of a CPR policy in the hospice. Semi-structured interviews were conducted on a purposive sample of hospice staff. The findings raise issues about the implementation of CPR in a context where it was felt that the patient should be allowed to die a natural death. PMID- 12734453 TI - Breaking bad news revisited: the push for negotiated disclosure and changing practice implications. AB - This article revisits the ethical, legal, professional and emotional issues involved with disclosing bad news. The authors examine the push for disclosure that has come from a number of quarters in the UK, including ethical and legal challenges, in particular the Bristol Royal Inquiry Report, professional codes of conduct, health policy and the expectations of the public. The contribution of nurses to breaking bad news is not widely discussed in the literature. With the development of new nursing roles and evidence-based practice it is timely to consider the role of nurses in this process. The article highlights some limitations with current guidelines for breaking bad news, in particular, that these guidelines tend to be constructed from a professional standpoint and lack patient-centred evidence. The issue of emotional labour and how it relates to giving bad news is discussed with respect to professional staff and patients. The article concludes by raising some practice implications, including: the importance of context and continuity; the significance of information and support; the desirable qualities of the professional; and issues to consider in determining patient preferences. PMID- 12734454 TI - Difficulties in identifying distress and its causes in people with severe communication problems. PMID- 12734455 TI - The PNP's role in supporting infant and family well-being during the first year of life. PMID- 12734456 TI - Let's make it an active summer. PMID- 12734457 TI - Supporting adolescent mothers and their children through a high school-based child care center and parent support program. AB - INTRODUCTION: A pilot study was conducted of adolescent parents and their children at a high school-based child care center to examine the influence of the program on parental competence, parent-child interaction, and child development. METHOD: A convenience sample of 18 urban adolescent mothers with children was interviewed at the end of one academic year. Mothers completed the Maternal Self Report Inventory (MSRI; short form) and the Parenting Daily Hassles Scale (PDHS). Parent-child teaching interactions were rated with the Nursing Child Assessment Teaching Scale (NCATS), and the Michigan Early Developmental Profile (MEDP) was used to evaluate child developmental outcomes. RESULTS: Mothers reported positive scores on the MSRI and PDHS. NCATS observations indicated ratings significantly higher when compared (assuming unequal variances) with a group matched for age range and length of time in school of adolescent mothers from the Nursing Child Assessment Satellite Training (NCAST) national database. The MEDP indicated that 77% of the children assessed were within normal limits for all domains (fine motor, cognition, language, gross motor, and self care). DISCUSSION: The results of this pilot study indicate that adolescent mothers and their young children in the sample benefited from the educational and support services offered at a school-based child care and parent support program. Further study is indicated to test the strength and duration of the effects. PMID- 12734458 TI - Parental perceptions of their child's asthma: management and medication use. AB - INTRODUCTION: Adequate treatment for asthma depends on accurate assessment and intervention by the parent and child and timely communication with the provider. These actions by the parent may be affected by their understanding of asthma management and their concerns about medications being prescribed. This research reports parental experiences with their children with asthma, specifically their beliefs, knowledge, and attitudes about asthma management, including medication use. METHODS: Data reported are from a study investigating parental attitudes and beliefs affecting antiinflammatory medication use in childhood asthma. These qualitative findings emerged from one-on-one semistructured qualitative interviews with 18 parents of children 2 to 18 years of age who were from diverse racial and socioeconomic backgrounds and who represented the spectrum of illness severity. RESULTS: Eight main themes within the domain of asthma management and medication use were identified: "I know my child," "trial and error," "partnership," "need for education," "negotiating responsibility," "hassles with medication administration," "preferences," and "the benefits outweigh the risks of side effects." DISCUSSION: These themes emphasize parents' need to partner with providers in their child's asthma management, as well as their need for ongoing asthma education. Parents also expressed concern about adverse effects of antiinflammatory medication but acknowledged the importance of controlling asthma symptoms. Based on these findings, systematic practice changes are recommended that provide regular opportunities for parent and child asthma education in a structured asthma wellness or "tune-up" visit. PMID- 12734459 TI - The risks and alternatives to physical punishment use with children. AB - Despite strong evidence of negative developmental outcomes resulting from the use of physical (or corporal) punishment with children, its use by parents and other caregivers is common. Such negative outcomes include child aggression, mental health issues, and physical abuse. Health care providers have a responsibility to promote disciplinary strategies that facilitate positive parent-children relationships and keep children's self-esteem and bodies healthy and intact. The incidence, factors, and outcomes associated with parental use of physical punishment are reviewed and useful advice for parents and age-appropriate disciplinary strategies and resources are outlined for the various stages of child development from infancy to school age. PMID- 12734460 TI - Employment characteristics of recent PNP graduates. AB - PURPOSE: The purpose of this study was to describe the characteristics of employment, health-illness characteristics of children cared for by PNPs, frequency of role functions by practice setting, and participation in professional organizations of recent graduates from 11 university programs. METHOD: A 60-item multiple-choice survey tool was sent to graduates from 1997 1999. RESULTS: Two hundred twenty-five surveys were returned (response rate: 54%). More than 48% described their practice setting as primary care, and 33% described it as specialty practice. Graduates indicated they frequently cared for children "at risk," with chronic health conditions, and with critical health conditions. Assessment of growth and development and provision of preventive care services were roles more frequently performed in primary care settings, whereas implementing and participating in research, case management, systems intervention, staff development, development of policies or programs, and writing for grants or publication were roles more frequently performed in specialty practice settings (P <.05). DISCUSSION: Findings from this survey confirm earlier survey findings regarding the varied practice settings, diverse role functions, and type of clients served by recent PNP graduates. Educational programs need to prepare graduates for the varied role functions they are likely to perform upon graduation and encourage them to participate in professional organizations and the development of health care policies affecting children and families. PMID- 12734461 TI - Sexual abstinence and the Sexual Abstinence Behavior Scale. AB - INTRODUCTION: Nurses may choose to promote sexual abstinence, particularly when they are working with young adolescents. However, it is difficult to know how to intervene because sexual abstinence has not been well defined in the literature. The purpose of this study was to evaluate the psychometrics of a four-item measure of sexual abstinence behavior (the SABS) that both registered nurses and advanced practice nurses may find useful in their practice. METHOD: African American, middle school students (n = 113) completed a self-administered questionnaire during their health education class. The questionnaire contained the SABS along with items assessing demographics, sexual behavior, and psychosocial variables related to sexual behavior. RESULTS: Psychometric analysis supported SABS reliability (alpha =.73) and validity. For example, SABS scores correlated with perceived negative consequences of teen sex (r = 0.38) and sexual abstinence self-efficacy (r = 0.48). DISCUSSION: Although the SABS is still in a preliminary stage of development, it is useful for assessment purposes and for directing registered nurses and advanced practice nurses toward specific behaviors they may want to promote when teaching sexual abstinence to their young adolescent patients. PMID- 12734462 TI - Diagnosis and management of pelvic inflammatory disease in adolescents. PMID- 12734463 TI - Whiplash injury secondary to a motor vehicle accident. PMID- 12734464 TI - Helping parents make sense of ADHD diagnosis and treatment. PMID- 12734465 TI - Population-based health care. PMID- 12734466 TI - Firearm safety devices. PMID- 12734468 TI - Effect of antianginal medication on resting myocardial perfusion and pharmacologically induced hyperemia. AB - Background. Patients scheduled for myocardial perfusion imaging are often taking several antianginal drugs. There is presently no consensus concerning a regimen of discontinuation before either rest or pharmacologic stress myocardial perfusion imaging. Whether antianginal treatment affects diagnostic sensitivity and specificity is not well documented. Methods and Results. The effect of the three most commonly used antianginal drugs (nitroglycerin, 400 micro g [NTG]; metoprolol, 50 mg [MET]; and amlodipine, 5 mg [AML]) on myocardial perfusion was tested in 49 patients (age, 63 +/- 8 years; 43 men) allocated prospectively to one of the treatments (NTG, n = 25; MET, n = 14; and AML, n = 10). All patients had documented coronary artery disease and were scheduled for elective percutaneous coronary intervention. Patients were studied once on treatment and once off treatment with an interval of 1 to 3 weeks. For NTG, the measurements were performed on the same day with an interval of 1 hour. The MET and AML groups were also studied during dipyridamole-induced hyperemia (0.56 mg. kg(-1). min(-1) for 4 minutes). So that a quantitative value of myocardial perfusion in milliliters per gram per minute could be obtained, myocardial perfusion was quantified with nitrogen 13 ammonia positron emission tomography as an average of the midventricular perfusion in each of the 3 vascular territories. NTG treatment increased the overall resting perfusion (0.75 +/- 0.18 vs 0.86 +/- 0.22, P <.05), whereas resting perfusion was reduced after MET treatment (0.92 +/- 0.14 vs 0.82 +/- 0.17, P <.05). AML treatment did not alter resting perfusion (0.87 +/- 0.22 vs 0.87 +/- 0.23, P = NS). Dipyridamole-induced hyperemia was reduced after treatment with MET (2.02 +/- 0.66 vs l.57 +/- 0.52, P <.001), whereas the hyperemic response was unchanged after treatment with AML (1.54 +/- 0.49 vs 1.86 +/- 0.91, P = NS). Conclusions. Antianginal medication can alter both resting and hyperemic myocardial perfusion and might affect the ability to detect flow limiting stenosis. NTG increases perfusion, MET reduces perfusion, and AML does not affect perfusion. Larger-scale trials are warranted to establish a consensus for optimal antianginal medication for patients undergoing perfusion imaging. PMID- 12734470 TI - Cutaneous mucinosis in dermatomyositis associated with a malignant tumor. AB - We report a case of dermatomyositis presenting with cutaneous mucinosis as the sole manifestation. A malignancy screen revealed an underlying nasopharyngeal carcinoma. PMID- 12734471 TI - Severe measles in a young female patient with chronic, generalized Trichophyton rubrum infection showing type 2 helper T cell-dominant immunologic reactivity. AB - An obese 18-year-old girl without a history of atopic dermatitis or a systemic immunosuppressive disorder had severe measles with liver dysfunction and pneumonia. For the previous 3 years she had had a chronic untreated generalized Trichophyton rubrum infection. The rash of the measles tended to spare the fungal lesions, where numerous fungi grew in the stratum corneum. Serum IgE level was markedly elevated (>16,000 IU/mL) and decreased after effective antifungal therapy. Delayed skin hypersensitivity reactions to trichophytin and tuberculin were negative, but the immediate skin hypersensitivity to trichophytin was positive. These data indicate that the patient had acquired hyperproduction of IgE with suppression of cell-mediated immunity, that is, acquired impairment of the balance between type 1 and type 2 helper T cells. In this case, chronic T rubrum infection was thought to have a role in driving a normal balance between type 1 and type 2 helper T cells toward type 2 helper T cell-dominant immunity. The result was deterioration of the measles infection and prolongation of the dermatophytosis. PMID- 12734472 TI - Massive exophytic abscesses and fibrotic masses of the chin: a variant of the follicular occlusion triad. AB - We present a patient with an extensive cluster of exophytic nodules that developed on his chin. These nodules consisted of abscesses and fibrotic areas. Lesion morphology, histology, and microbiology support a follicular occlusion triad entity. However, the distribution is striking and does not fit the entities described in the triad. We present the case to show that follicular occlusion was the inciting factor in our patient's eruption and to broaden our concept of clinical manifestations that can arise from this pathologic process. PMID- 12734473 TI - Dermal leiomyosarcoma. AB - Leiomyosarcoma of the skin is a rare tumor that cannot be diagnosed clinically because its gross appearance is nonspecific. The primary skin tumor usually appears as a solitary nodule. It may arise de novo in the dermis or in the subcutaneous tissue, or as a metastatic lesion from an internal neoplasm. Dermal leiomyosarcomas rarely metastasize, unlike the subcutaneous tumors. We describe a case of the dermal type of leiomyosarcoma. PMID- 12734474 TI - Dermatofibrosarcoma protuberans occurring in a smallpox vaccination scar. AB - Dermatofibrosarcoma protuberans (DFSP) is a locally aggressive, rarely metastatic, spindle cell tumor. Trauma has been associated with its development. Since the 1940s, malignant tumors have been described to occur in sites of smallpox vaccination scars. Five cases in the literature document DFSP arising in sites of prior immunizations. We report a case of DFSP occurring in a smallpox vaccination scar and review the available literature. PMID- 12734475 TI - Vancomycin-associated linear IgA bullous dermatosis mimicking toxic epidermal necrolysis. AB - Vancomycin is the most frequent trigger of drug-induced linear IgA bullous dermatosis. We describe a fulminant case of linear IgA bullous dermatosis in a 74 year-old man who experienced skin sloughing of 90% of his body surface after receiving vancomycin. PMID- 12734476 TI - Jogger's toenail. AB - Sports-related toenail abnormalities result from trauma during specific athletic events. A case is presented of jogger's toenail that developed in a patient who had just begun a running program. The cause, diagnosis, and management of jogger's toenail are discussed. Knowledge of myriad toenail disorders associated with sporting events helps the clinician to avoid misdiagnosis. PMID- 12734477 TI - Herpes simplex virus type 2-associated eosinophilic cellulitis (Wells' syndrome). AB - Eosinophilic cellulitis (Wells' syndrome) is a recurrent inflammatory dermatosis characterized by massive infiltration of eosinophils into the skin. Drugs and pathogens have been recognized causes of eosinophilic cellulitis. We report the repeated association of eosinophilic cellulitis with herpes simplex virus type 2 infections. Antiviral therapy led to a complete remission of eosinophilic cellulitis, indicating that causative treatment of underlying diseases can be effective in controlling eosinophilic cellulitis. PMID- 12734478 TI - Extramedullary hematopoiesis in an adult. AB - Cutaneous extramedullary hematopoiesis is a rare disease, affecting mainly children with intrauterine viral or hematologic disorders. To our knowledge, only about 30 cases have been reported in adults, mainly associated with myelofibrosis. We report a new case of this entity in a 79-year-old woman and comment on the clinical and histologic differential diagnosis. PMID- 12734479 TI - Basal cell nevus syndrome concurrent with adenoid cystic carcinoma of salivary gland. AB - Basal cell nevus syndrome is a genetically determined disease characterized by 5 major manifestations (multiple basal cell carcinomas, jaw cysts, skeletal abnormalities, pits of the hands and feet, and ectopic calcification) and a variety of developmental anomalies. We present a case of basal cell nevus syndrome in which the patient had adenoid cystic carcinoma of the minor salivary glands develop in the soft palate resulting in distant pulmonary metastases. The patient died at the age of 44 years of respiratory complications. To our knowledge, this is the first simultaneous occurrence of basal cell nevus syndrome with adenoid cystic carcinoma of salivary gland. PMID- 12734480 TI - Giant acquired digital fibrokeratoma. AB - We describe a 33-year-old patient with a giant skin tumor on her right big toe. The tumor was characterized by unusual clinical manifestations including huge size and papillomatous surface. Chronic long-standing irritation as a result of aerobic exercise may have been involved in the development of giant fibrokeratoma with papillomatous surface. PMID- 12734481 TI - Paraneoplastic pemphigus associated with uterine carcinoma. AB - Paraneoplastic pemphigus (PNP) is an autoimmune blistering disease characterized by severe mucous membrane involvement, polymorphous skin eruptions, and underlying neoplasms, usually those of lymphoreticular system. Cases of PNP associated with solid cancer are extremely rare. A 46-year-old woman presented with severe stomatitis, pseudomembranous conjunctivitis, and polymorphous skin eruptions. Histologic, immunofluorescence, and immunoblot analyses confirmed the diagnosis of PNP. Whole-body examination revealed squamous cell carcinoma of the uterine cervix, but no hematologic tumors. Complete removal of the uterine tumor in addition to administration of betamethasone induced considerable improvement of cutaneous and mucous membrane lesions. To our knowledge, this is the first reported case of PNP associated with uterine carcinoma. PMID- 12734482 TI - A tattooed butterfly as a vector of atypical Mycobacteria. AB - We report the first case of cutaneous inoculation of atypical Mycobacteria secondary to tattooing. The diagnosis of atypical Mycobacteria infection of the skin was confirmed on the basis of the clinical and histologic appearance, the detection of acid-fast bacilli on Ziehl-Neelsen stain, and positive polymerase chain reaction. The medical complications of tattooing, which are manifold, are briefly summarized. This case emphasizes the need for federal regulation of tattooing, which is an invasive procedures associated with infectious and noninfectious complications. PMID- 12734483 TI - Congenital self-healing reticulohistiocytosis presenting with hemorrhagic bullae. AB - Congenital self-healing reticulohistiocytosis is a variant of Langerhans cell histiocytosis. It is present at birth or appears in the neonatal period and involutes spontaneously within 3 to 4 months. Although the skin eruptions in most cases consist of papulonodules, several patients with vesicular or bullous lesions have been reported. We describe a case of congenital self-healing reticulohistiocytosis presenting hemorrhagic bullae that mimicked hemangioma. PMID- 12734484 TI - Erythema multiforme in a neonate. AB - We describe a case of erythema multiforme in a 2-week-old boy. He had no remarkable antecedents, and a polymerase chain reaction-based technique failed to detect herpes simplex virus DNA in the skin biopsy specimen. To our knowledge, only one previous biopsy-proven case of erythema multiforme during the neonatal period has been reported. PMID- 12734485 TI - Confluent and reticulated papillomatosis: response to tazarotene. AB - Confluent and reticulated papillomatosis of Gougerot and Carteaud is an uncommon dermatosis of unclear cause that can be recalcitrant to therapy. We report the case of an 11-year-old black girl with the eruption in whom it completely cleared using tazarotene gel. The treatment was well-tolerated and we suggest considering topical tazarotene as an alternative to systemic retinoid therapy. PMID- 12734486 TI - Cutaneous marginal zone B-cell lymphoma: a case accompanied by massive plasmacytoid cells. AB - A 36-year-old man presented with a red nodule on his left shoulder. Histologically, there were variously sized, irregularly shaped nests throughout the dermis partly extending into the subcutaneous tissue. Masses of centrocyte like cells were situated in the center of the tumor nests and accompanied by adjacent secondary follicle structures. Partial follicular colonization was seen. Massive plasmacytoid cells were located in the papillary dermis and the periphery of the tumor nests. Immunohistochemically, these centrocyte-like cells were positive for CD19 and alkaline phosphatase, and negative for CD5 and CD10. Cytoplasm of the plasmacytoid cells was positive for IgG and lambda-light chain, and negative for IgM, IgA, and kappa-light chain. Monotypic immunoglobulin staining including light chain restriction was shown. Clonal immunoglobulin heavy chain gene rearrangement by Southern blot analysis was shown in the tumor tissue. Morphologic, immunohistochemical, and molecular studies revealed that this patient had a cutaneous marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue type. Electron beam (total 40 Gy) irradiation was applied. The tumor disappeared completely. Neither local recurrence or metastasis have appeared during 3 years of follow-up. PMID- 12734487 TI - Anetoderma: another facet of Lyme disease? AB - Lyme disease has been suspected to be one cause of secondary anetoderma. We present a 25-year-old male patient with multiple lesions of anetoderma with a diameter of up to 2 cm that developed within the last 4 weeks without subjective symptoms. The histopathologic pattern was similar to the inflammatory stage of acrodermatitis chronica atrophicans. Polymerase chain reaction analysis out of the paraffin-embedded tissue, confirmed by sequencing of the obtained nucleotide product, revealed a part of the 23 S ribosomal RNA gene of Borrelia burgdorferi sensu lato. Enzyme-linked immunosorbent assay showed an increased serum IgG titer against B burgdorferi corroborated by Western blot analysis. After a treatment with oral doxycycline (100 mg twice a day) for 30 days the anti-B burgdorferi titer decreased significantly and no new lesions appeared. Some cases of anetoderma might be caused by Borrelia and patients with anetoderma should be examined for borreliosis including serology and polymerase chain reaction of lesional skin. In cases of Borrelia-induced anetoderma, early antibiotic treatment could prevent further progression of the disfiguring skin lesions and the underlying disease. PMID- 12734488 TI - Thalidomide for treatment of severe generalized discoid lupus lesions in two patients with systemic lupus erythematosus. AB - We describe 2 patients with systemic lupus erythematous whose widespread discoid lupus erythematosus was unresponsive to systemic steroids and antimalarial agents. They showed dramatic improvement to thalidomide at a dose of 300 mg/d, with maximum benefit achieved within 15 weeks of therapy. Dosages of 50 to 100 mg/d were effective in maintaining remission for 1 year. However, thalidomide induced neuropathy was observed in both cases. PMID- 12734489 TI - Parasitism of basal cell carcinoma by lentigo maligna melanoma. AB - The coexistence of 2 types of malignant neoplasms is relatively uncommon. We report a pigmented melanocytic lesion occurring adjacent to a previously treated basal cell carcinoma. Histology showed melanoma (lentigo maligna type) colonizing basal cell carcinoma demonstrated by hematoxylin-eosin and immunocytochemistry stains. It is unclear whether this lesion has the metastatic potential of an invasive melanoma of similar thickness, or simply reflects melanoma in situ extending into an epidermal-derived tumor analogous to appendageal extension of lentigo maligna. We review the literature on melanoma colliding with basal cell carcinoma. Therapeutic and prognostic problems are posed by this unique situation. PMID- 12734490 TI - Bullous aplasia cutis congenita. AB - A tense yellow vesicle was noted on the back parietal scalp of a female newborn. This proved to be a bullous variant of aplasia cutis congenita. Only 16 cases of this apparently rare disorder have been previously reported. Histologic evaluation of such lesions reveals a distinct pattern containing fibrovascular stromas, edematous stroma, or both. Identical histologic findings are found in encephaloceles and meningoceles, supporting the recently proposed hypothesis that this variant of aplasia cutis may represent the form fruste of a neural tube closure defect. This disorder should be included in the differential diagnosis of vesicobullous lesions in the neonate. Bullous aplasia cutis congenita is a rare clinical subtype of aplasia cutis congenita with distinctive histologic findings. We present a new case, and summarize the clinical and histologic findings of the 16 cases previously reported in the English-language literature. Bullous or membranous aplasia cutis congenita may represent a form fruste of a neural tube defect. PMID- 12734491 TI - Juvenile sarcoidosis after BCG vaccination. AB - We report the case of a 2-year-old boy with juvenile sarcoidosis, in whom the cutaneous lesions first arose at the site of and soon after a BCG vaccination. Juvenile sarcoidosis is rare, and the pattern of clinical features is distinct from the adult form of sarcoidosis, possibly related to immunologic development. The cause of sarcoidosis is unknown, although there is much interest in the possibility of mycobacterial species operating as antigenic stimuli to initiate the disease. This case suggests that the Mycobacterium bovis present in the BGC vaccination may have been etiologically important in the development of sarcoidosis. PMID- 12734492 TI - Progesterone receptor-positive eruptive syringoma associated with diabetes. AB - Syringoma is a benign eccrine tumor that generally forms asymptomatic papules on facial skin. Eruptive forms of the tumor are uncommon and of uncertain cause. We report a case in which eruptive syringoma with clear cell morphology was found to express progesterone receptors, suggesting the possibility of hormonal control of the tumor. PMID- 12734493 TI - Autoimmune thyroid diseases: etiology, pathogenesis, and dermatologic manifestations. AB - Autoimmune thyroid disease (AITD) including Graves' disease, Hashimoto's thyroiditis, and idiopathic hypothyroidism (atrophic Hashimoto's thyroiditis) is of vital concern to the dermatologist. This article reviews the cutaneous manifestations of Graves' disease and Hashimoto's thyroiditis. Recognition of dermatologic manifestations of AITD may alert practitioners to investigate for these disorders. The immune response involved in the pathogenesis of AITD is detailed. Current understanding of the role of genetic and environmental factors, antigens, and apoptosis are elaborated. The future holds exciting insight into the etiology, pathogenesis, and treatment of AITD. PMID- 12734494 TI - Early melanoma detection: nonuniform dermoscopic features and growth. AB - BACKGROUND: Dermoscopy alone is not sufficient to detect all early melanomas. Total body photos reveal growth of melanomas but also reveal growth of melanocytic nevi. OBJECTIVE: We set out to determine whether a simplified algorithm on the basis of nonuniform dermoscopic features combined with growth noted from baseline total body photos targeted the early melanocytic neoplasms most likely to be malignant. METHODS: Lesions removed during follow-up of patients with total body photographs were reviewed and 169 melanocytic lesions were identified for which both gross clinical and dermoscopic photos were available. The images were evaluated separately by 3 academic dermatologists, without knowledge of the given pathologic diagnosis, for uniformity (consistent gradient of features from center to edge) and change (specifically, that which could indicate melanoma growth in normal skin or within a nevus). RESULTS: Using a minimum of 2 out of 3 agreement for uniformity and change, 12 of 16 melanomas (including all 5 superficially invasive tumors) were graded as nonuniform and changed. The 4 melanomas not included in this category were in situ. The predicted odds of melanoma for lesions scored as both nonuniform and changed was 4.06 (P >.0195). If 3 out of 3 agreement was used, the odds ratio increased to 6 (P >.0010). CONCLUSION: An algorithm on the basis of dermoscopic nonuniformity and change suggestive of growth as determined by total body photography segregates melanocytic neoplasms most likely to be malignant. PMID- 12734495 TI - Modified ABC-point list of dermoscopy: A simplified and highly accurate dermoscopic algorithm for the diagnosis of cutaneous melanocytic lesions. AB - BACKGROUND: The use of dermoscopy (epiluminescence microscopy, surface microscopy, dermatoscopy) improves clinical diagnostic sensitivity by 10% to 27%, particularly achieved by different algorithms or scores. OBJECTIVE: We sought to develop a simplified and highly accurate dermoscopic-point list for cutaneous melanocytic lesions. METHOD: We studied consecutive patients with suspicious melanocytic lesions, which were excised and histopathologically examined at our institution. On the basis of the ABCD rule of Stolz, Menzies score, and the modified ABCD rule of Kittler, a simplified ABC-point list was developed. Simple points were given for the following: asymmetry of outer shape (A) or differential structures inside the lesion in at least 1 axis ((A)); the abrupt cutoff of network at the border in at least one quarter of circumference (B); 3 or more colors (C); 3 or more differential structures (D); or noticed change (evolution) in the last 3 months (E). Using 20-fold magnification of computer dermoscopy, the sensitivity, specificity, and diagnostic accuracy were examined in 269 cutaneous melanocytic lesions. Of these, 84 (31.2%) were cutaneous melanomas. Also, the sensitivity, specificity, and diagnostic accuracy were investigated with a 7 point checklist and the 7 features for melanoma. RESULTS: With the ABC-point list for the diagnosis of cutaneous melanoma, sensitivity was 90.5%, specificity was 87%, and diagnostic accuracy was 88.1%, confirmed by cross-validation. The ABCD rule resulted in 90.5%, 72.4%, and 78.1%; Menzies score in 95.2%, 77.8%, and 83.3%; 7-point checklist in 90.5%, 87%, and 88.1%; and 7 features for melanoma in 94%, 74.6%, and 80.7%, respectively, CONCLUSIONS: The ABC-point list is simpler than the already established algorithms. Despite its simplicity, a high sensitivity, specificity, and diagnostic accuracy was achieved. This simplified approach in dermoscopic diagnostics may contribute to further spread and enable to learn and use this method more easily. PMID- 12734497 TI - Nodular melanoma: patients' perceptions of presenting features and implications for earlier detection. AB - BACKGROUND: The incidence of thick melanoma and related mortality is largely static despite advances in early detection during the last 20 years. Nodular melanoma (NM) accounts for the majority of thick lesions and is difficult to recognize in the early stages of its evolution. OBJECTIVE: The purpose of this study was to identify historic or clinical features that may facilitate earlier detection of NM. METHODS: A questionnaire was administered to 125 patients attending the Victorian Melanoma Service between 1998 and 2000 with superficial spreading melanoma or NM. Parameters were compared by tumor type and thickness. RESULTS: NMs are more often symmetric, elevated, uniform in color, and nonpigmented. Color change is uncommon. CONCLUSION: NM often fails to fulfill the ABCD diagnostic criteria. Biopsy after a set period of observation should aid differentiation from inflammatory lesions and enable earlier detection of this subtype. PMID- 12734496 TI - Dermoscopy of pigmented skin lesions: results of a consensus meeting via the Internet. AB - BACKGROUND: There is a need for better standardization of the dermoscopic terminology in assessing pigmented skin lesions. OBJECTIVE: The virtual Consensus Net Meeting on Dermoscopy was organized to investigate reproducibility and validity of the various features and diagnostic algorithms. METHODS: Dermoscopic images of 108 lesions were evaluated via the Internet by 40 experienced dermoscopists using a 2-step diagnostic procedure. The first-step algorithm distinguished melanocytic versus nonmelanocytic lesions. The second step in the diagnostic procedure used 4 algorithms (pattern analysis, ABCD rule, Menzies method, and 7-point checklist) to distinguish melanoma versus benign melanocytic lesions. kappa Values, log odds ratios, sensitivity, specificity, and positive likelihood ratios were estimated for all diagnostic algorithms and dermoscopic features. RESULTS: Interobserver agreement was fair to good for all diagnostic methods, but it was poor for the majority of dermoscopic criteria. Intraobserver agreement was good to excellent for all algorithms and features considered. Pattern analysis allowed the best diagnostic performance (positive likelihood ratio: 5.1), whereas alternative algorithms revealed comparable sensitivity but less specificity. Interobserver agreement on management decisions made by dermoscopy was fairly good (mean kappa value: 0.53). CONCLUSION: The virtual Consensus Net Meeting on Dermoscopy represents a valid tool for better standardization of the dermoscopic terminology and, moreover, opens up a new territory for diagnosing and managing pigmented skin lesions. PMID- 12734498 TI - Lymph node grading for staging of mycosis fungoides may benefit from examination of multiple excised lymph nodes. AB - BACKGROUND: Lymph node (LN) histopathologic class has been shown to be a significant determinant of survival in patients with mycosis fungoides. Often, histopathologic evaluation of just 1 node is used in staging patients with cutaneous T-cell lymphoma. OBJECTIVE: We examined whether sampling multiple nodes alters the staging and prognostic group placement of patients with mycosis fungoides as compared with sampling just 1 node. METHODS: Multiple LNs were obtained from a single, local region for histopathologic evaluation and grading in 8 patients with mycosis fungoides. RESULTS: Differences in histopathologic grading using multiple nodes were found in 5 of 8 patients. There was a potential upstaging of the assigned disease stage, compared with the stage that might have been assigned had just 1 node been sampled, in 3 patients. The differences in LN grading also potentially led to differences in prognostic group placement in 4 patients. CONCLUSION: Determining histopathologic grades from multiple LNs may allow a more accurate stage and prognosis to be assigned to patients. PMID- 12734499 TI - Numerous, small, darkly pigmented melanocytic nevi: the cheetah phenotype. AB - BACKGROUND: The presence of multiple atypical nevi or numerous melanocytic nevi increases the risk for the development of cutaneous melanoma. OBJECTIVE: We sought to describe a distinct clinical phenotype characterized by numerous (>100), small (< or =4 mm), darkly pigmented melanocytic nevi that are uniform in color. METHODS: Biopsy specimens from 6 patients (3 men and 3 women; age range, 44 to 81 years) with this clinical phenotype were reviewed and compared with a database of melanocytic lesions analyzed by the Yale Dermatopathology Laboratory (YDL) in the year 2000. RESULTS: Of the 6 patients, 4 had multiple primary melanomas develop (n = 2-4), ranging from in situ to 1.0 mm in depth. The other 2 patients each had 1 nevus with severe cytologic atypia. When compared with the YDL database, our patients were more likely to have the following pigmented lesions: junctional melanocytic nevi, junctional lentiginous nevi, junctional nevi with cytologic atypia, and simple lentigines (P <.001). CONCLUSIONS: The longitudinal evaluation of patients with this phenotype can be challenging because similar-appearing pigmented lesions (small and uniformly dark-brown to black) had a range of histologic diagnoses from simple lentigo to junctional lentiginous nevus to thin melanoma. PMID- 12734500 TI - Analysis of patients with suspected photosensitivity referred for investigation to an Australian photodermatology clinic. AB - BACKGROUND: Australia's first dedicated photodermatology clinic was established at St Vincent's Hospital Melbourne in 1993. OBJECTIVE: We sought to review clinical diagnoses and results of investigations performed on patients seen at this institution. METHODS: A database was created to enable a retrospective and prospective analysis of all patients attending for evaluation of suspected photosensitivity from April 1993 to October 2000. RESULTS: A total of 513 patients were seen, 289 (56.3%) female and 224 (43.7%) male, with a mean age of 45.2 years (range: 2.6-85.9). A photosensitive disorder was diagnosed in 397 patients (77.4%), with the acquired idiopathic photodermatoses accounting for diagnoses in 215 (41.9%) of all patients seen. The most common diagnosis was polymorphous light eruption. Reduced minimal erythema doses were seen in 25.3% of all patients light tested. In those photopatch tested, 23.3% had at least 1 photocontact reaction. Allergic contact dermatitis in a photosensitive distribution was diagnosed in 7.4% of the clinic population. CONCLUSION: A large proportion of referrals to a photodermatology clinic comprise people with acquired idiopathic photodermatoses, with other common diagnoses that may mimic photosensitivity including allergic contact dermatitis, atopic dermatitis, and rosacea. PMID- 12734501 TI - Histopathologic evaluation of cutaneous squamous cell carcinoma: results of a survey among dermatopathologists. AB - BACKGROUND: There are numerous histopathologic features related to prognosis in cutaneous squamous cell carcinoma (CSCC). We hypothesize that there is no uniform approach toward the reporting of these features. This may be related to differing opinions on their prognostic use. METHODS: A written survey concerning the microscopic evaluation of CSCC was sent to 120 dermatopathologists in the United States and Canada. Respondents were asked whether they comment on specific microscopic features of CSCC, and whether they believe that each specific feature can predict prognosis. RESULTS: The response rate was 78%. Histologic type, and the presence of perineural or vascular/lymphatic invasion, is reported by most dermatopathologists (90%, 96%, and 95%, respectively). These features are also thought to predict prognosis by the majority of respondents. Only 54% report histologic grade, and 49% think grade predicts prognosis. Depth is reported anatomically by 63%, but by only 8% in actual millimeters of invasion. However, 55% think tumor depth predicts prognosis. A total of 43% report the presence of an associated actinic keratosis, although very few (16%) think it predicts prognosis. Very few comment on the presence of inflammation. CONCLUSIONS: Histopathologic reporting of CSCC is not uniform among dermatopathologists. Also, there appears to be differing opinions on the use of certain histopathologic features for predicting prognosis. PMID- 12734502 TI - Confocal histopathology of irritant contact dermatitis in vivo and the impact of skin color (black vs white). AB - BACKGROUND: The pathogenesis of irritant contact dermatitis and its modulation according to skin color is not well understood. Reflectance confocal microscopy (RCM) enables high-resolution, real-time, in-vivo imaging of human skin. OBJECTIVE: The goal of our study was to use RCM to determine whether susceptibility to irritant contact dermatitis differs between black and white skin. METHODS: Participants were placed in groups on the basis of skin color and the volar aspects of their forearms exposed to 1% and 4% sodium lauryl sulfate using Finn Chambers (Allerderm Laboratories Inc, Petaluma, Calif). They were evaluated at 6, 24, and 48 hours by RCM, transepidermal water loss, laser Doppler velocimetry, and routine histology. RESULTS: Participants with white skin had more severe clinical reactions than those with black skin. RCM revealed microscopic changes even without clinical evidence of irritation. Confocal features included parakeratosis, spongiosis, perivascular inflammatory infiltrate, and microvesicle formation, and these features were confirmed by routine histology. Also, participants with white skin had greater mean increases in transepidermal water loss after exposure to 4% sodium lauryl sulfate than did participants with black skin. CONCLUSION: In-vivo RCM can track early pathophysiologic events revealing differences between black and white skin during the development of irritant contact dermatitis, and may support the theory that those with black skin are more resistant to irritants. PMID- 12734504 TI - Assessment of dermatopharmacokinetic approach in the bioequivalence determination of topical tretinoin gel products. AB - BACKGROUND: A new dermatopharmacokinetic (DPK) approach has been proposed for bioequivalence determination of topical drug products by comparing the drug content kinetics in stratum corneum. OBJECTIVE: We sought to establish any correlation between clinical safety/efficacy and DPK approach in bioequivalence determination of tretinoin gel 0.025%. METHODS: Tretinoin and isotretinoin were quantified in human volar forearm stratum corneum as a function of time with 3 tretinoin gel 0.025% products in 49 patients. Stratum corneum layers were harvested using multiple adhesive disks, which were subsequently extracted and quantified for both isomers by high-performance liquid chromatography. RESULTS: Products with similar composition and therapeutic equivalence were found bioequivalent, and products with different composition and clinical profiles were found bioinequivalent by DPK methodology. CONCLUSIONS: There is a direct correlation between DPK parameters in healthy patients and clinical safety/efficacy of tretinoin gel products in patients with acne. Data support the use of DPK parameters and methodology in the bioequivalence assessment of topical tretinoin gel products. PMID- 12734503 TI - Vulvar syringoma: a clinicopathologic and immunohistologic study of 18 patients and results of treatment. AB - BACKGROUND: Syringoma of the vulva has been rarely reported. No effective treatment modality has been documented. OBJECTIVE: The purpose of this study was to describe the clinicopathologic features of vulvar syringoma, to investigate the hormonal influence on its growth, and to establish an effective treatment modality. METHODS: A total of 18 cases of vulvar syringoma were selected from the surgical pathologic file at Chang Gung Memorial Hospital. In all, 15 formalin fixed, paraffin-embedded specimens were examined by immunohistochemical staining for estrogen receptor and progesterone receptor. RESULTS: The age of first presentation at our vulvar clinic ranged from 21 to 60 years with a median age of 29.5 years. Of patients, 13 (72%) had vulvar pruritus and 7 noticed aggravation during summer or during menstruation. The most common clinical appearance was multiple flesh-colored or brownish papules on bilateral sides of the vulva (9 of 18). One third of our series was found to have coexisting eyelid syringoma and 4 of them also had a family history of periorbital syringoma. Immunohistochemical stainings for estrogen receptor and progesterone receptor were all negative on the 15 cases studied. Of our patients, 7 with intense pruritus were treated with carbon dioxide laser vaporization. Their lesions resolved and pruritus subsided. CONCLUSION: Vulvar syringoma is not very rare and should be considered in the differential diagnosis of vulvar pruritus. In our study, estrogen receptor and progesterone receptor were not detected in vulvar syringoma. Carbon dioxide laser was an effective therapeutic modality in treating patients with intractable symptoms. PMID- 12734505 TI - Transplants from balding and hairy androgenetic alopecia scalp regrow hair comparably well on immunodeficient mice. AB - Human hair follicles were grafted onto 2 strains of immunodeficient mice to compare the regeneration potential of vellus (miniaturized, balding) and terminal (hairy, nonbalding) follicles from males and a female exhibiting pattern baldness. Each mouse had transplants of both types of follicles from a single donor for direct comparison. Grafted follicles from 2 male donors resulted in nonsignificant differences in mean length (52 mm vs 54 mm) and mean diameter (99 microm vs 93 microm) at 22 weeks for hairs originating from balding and hairy scalp, respectively, corresponding to 400% versus 62% of the mean pretransplantation diameters. Follicles from the female donor transplanted to several mice also resulted in nonsignificant differences in length (43 mm vs 37 mm) for hairs from balding and hairy scalp, respectively, during a period of 22 weeks. The mean diameter of the originally vellus hairs increased 3-fold, whereas the terminal hairs plateaued at approximately 50% of pretransplantation diameter, resulting in a final balding hair volume double that of the nonbalding hairs. This report shows that miniaturized hair follicles of pattern alopecia can quickly regenerate once removed from the human scalp and can grow as well as or better than terminal follicles from the same individual. PMID- 12734506 TI - Tazarotene cream in the treatment of psoriasis: Two multicenter, double-blind, randomized, vehicle-controlled studies of the safety and efficacy of tazarotene creams 0.05% and 0.1% applied once daily for 12 weeks. AB - BACKGROUND: Tazarotene in a gel formulation is widely used in the treatment of psoriasis. OBJECTIVE: To determine the efficacy and safety of tazarotene 0.1% and 0.05% creams in the treatment of psoriasis. METHODS: A total of 1303 patients participated in 2 clinical trials. Patients applied tazarotene creams 0.1% and 0.05% or vehicle once daily to all psoriatic lesions for 12 weeks followed by a 12-week posttreatment period. RESULTS: Both creams were significantly more effective than vehicle on the basis of an overall assessment of psoriasis, a global response to treatment, and reduction in plaque elevation and scaling. Therapeutic effect was maintained during the posttreatment period. Common adverse events included signs and symptoms of skin irritation. CONCLUSION: Tazarotene creams were associated with significant reductions in the severity of the clinical signs of psoriasis and were found to be safe with acceptable tolerability. Tazarotene cream 0.1% was generally more effective, although slightly less well tolerated, than the 0.05% cream. PMID- 12734507 TI - The new 940-nanometer diode laser: an effective treatment for leg venulectasia. AB - BACKGROUND: The 940-nm diode laser has been shown to be an effective treatment for leg veins. OBJECTIVE: We sought to evaluate the effectiveness of the 940-nm diode laser on leg veins, depending on the size and morphologic aspect of the treated vessels. METHODS: A total of 60 patients (mean age: 44.4 years, Fitzpatrick skin types I-IV) underwent up to 3 treatment sessions at 4-week intervals using the 940-nm diode laser. Treatment parameters were: vessels < 0.4 mm in diameter, 0.5-mm spot, pulse duration of 10 milliseconds, fluence 306 J/cm(2); 0.4 to 0.8 mm in diameter, 1-mm spot, pulse duration of 30 milliseconds, fluence 306 J/cm(2); and 0.8 to 1.4 mm in diameter, 1.5-mm spot, pulse duration of 70 milliseconds, fluence 317 J/cm(2). Repetition rate was 2.5 Hz. Success rate was evaluated through double-blind observation. RESULTS: Only 13.33% of patients with telangiectases less than 0.4 mm in diameter had a percentage of vessel clearance superior to 75%. However, 88.24% of patients with vessels between 0.8 and 1.44 mm in diameter obtained more than 75% vessel clearance. CONCLUSION: The treatment of leg veins by the 940-nm diode laser strongly depends on the size of the target vessel. Better results were obtained with 0.8- to 1.4-mm leg venulectases. PMID- 12734508 TI - Long-term implications of T-cell receptor gene rearrangement analysis by Southern blot in patients with cutaneous T-cell lymphoma. AB - BACKGROUND: T-cell clonality analysis by Southern blot (TSB) in skin biopsy specimens suggestive of mycosis fungoides may be helpful in confirming the diagnosis of a cutaneous lymphoma. However, there are no data available regarding the long-term prognostic implication of such results. OBJECTIVES: We sought to determine the long-term prognostic significance of TSB results from skin biopsy specimens of patients with mycosis fungoides. METHODS: We reviewed the records from the Cleveland Clinic Foundation and Northwestern University Medical Center for cases of biopsy-proven mycosis fungoides with results available for skin biopsy TSB from 1987 to 1990. RESULTS: The detection of clonality by TSB correlates with a higher TNM stage (median stage for positive TSB, IIb vs negative TSB, Ib; P <.05), but not with age at presentation (62 vs 59 years) or duration of disease before presentation (6.2 vs 5.9 years). Although the long term survival was not significantly different between the 2 groups, there was a trend for patients with positive TSB to die earlier (5-year survival of 67% vs 87%). Disease progression did not correlate with TSB results. Higher clonality rates were noted among patients with biopsy specimens showing a denser lymphoid infiltrate and a higher grade of cytologic atypia. CONCLUSIONS: Detection of clonality with TSB requires a significant clonal burden. Although clonality can be detected in patients with patches and plaques (T1 and T2) most cases with positive results were obtained from patients with advanced disease (T3 and T4). In our experience, detection of clonality by TSB does not correlate with disease progression and does not carry long-term prognostic implications. PMID- 12734509 TI - Malignant melanoma. PMID- 12734510 TI - The International League of Dermatological Societies: from the World Congress of Dermatology to the International Foundation for Dermatology and beyond. PMID- 12734511 TI - Surgical pearl: Scalpel dermabrasion complements shave excision. PMID- 12734512 TI - Surgical pearl: Nail plate separation and splint fixation--a new noninvasive treatment for pincer nails. PMID- 12734514 TI - Prevalence of Becker's nevi in a population of young men in central Italy. PMID- 12734515 TI - The myth of Pautrier's microabscesses. PMID- 12734516 TI - Potentially fatal interstitial lung disease can occur in clinically amyopathic dermatomyositis. PMID- 12734517 TI - Infection control practitioners' perceptions and educational needs regarding bioterrorism: results from a national needs assessment survey. AB - BACKGROUND: The perceived threat that biological weapons will be used in an act of terror against the United States has escalated sharply since the discovery of anthrax-tainted letters after the terrorist attacks of September 11, 2001. These events underscore the critical nature of health care and public health preparedness and the need to augment infection control practitioner education and training. METHODS: Between October 2000 and August 2001 a national needs assessment was conducted by use of a 35-question survey. The survey measured infection control practitioners' (ICPs') perception of the risk for bioterrorism in the United States and in their community, the proportion of ICPs with prior training in bioterrorism preparedness, and preferences for delivery media of future bioterrorism education. RESULTS: The assessment of the perceived threat of bioterrorism in the United States during the next 5 years (P =.022) and in the ICPs' work community (P <.001) revealed significant regional differences. Only half (56%) of the respondents reported prior training in bioterrorism preparedness. Respondents reported that the 2 most common barriers to receiving training were lack of training opportunities (70.2%) or no dedicated work time for training (19.4%). CONCLUSIONS: The results of this study indicate an urgent need for more resources and opportunities for clinical education in bioterrorism preparedness that will provide continuing education credit. Successful bioterrorism education will require a variety of instructional designs and media delivery methods to address ICPs' preferences and needs. PMID- 12734518 TI - Microbiologic profile of intra-abdominal infections at Belo Horizonte, Brazil. AB - Intra-abdominal infections (IAIs) represent one of the most common clinical problems in hospital practice, especially in surgical areas and centers of intensive care. The treatment of IAIs generally involves the draining of abscesses and empirical antimicrobial therapy. In this study, among 150 patients suffering from IAI, 106 (70.7%) yielded samples that presented microbial growth. Polyinfection was detected in 51.9% of the cases and varied from 2 to 9 distinct microbes per specimen. The overall mean number of micro-organisms isolated per patient was 2.17. Aerobic bacteria (as strict aerobes and facultative anaerobes), strict anaerobic bacteria, and fungi of the genus Candida represented 93.4%, 30.2%, and 13.2% of the cases positive for micro-organisms, respectively. The most common aerobic bacteria were those of the genera Staphylococcus, Escherichia, Proteus, and Streptococcus. Despite the frequent prior use (52%) with antimicrobials of recognized action against strict anaerobes, these micro organisms constituted 30.9% of the total isolates, and the most frequently found were of the Bacteroides fragilis group and Prevotella species. The high prevalence of anaerobes in the specimens obtained from IAI demonstrates the need to give greater importance to these micro-organisms by making available material and human resources to carry out culture of the anaerobes as part of routine hospital procedures. PMID- 12734519 TI - Do we practice what we preach? Health care worker screening and vaccination. AB - OBJECTIVE: To describe the current screening and immunization practices in New South Wales (NSW) hospitals and the experience of NSW nurses in relation to screening and immunization and to identify areas that can be targeted for improvement. DESIGN: This was a cross-sectional survey. SETTING: The study was performed in NSW, Australia. METHODS: We used a written questionnaire to survey the infection control/occupational health coordinators of all of the 85 private hospitals and 204 eligible public hospitals in NSW and 800 randomly sampled registered nurses. RESULTS: Response rates were high (hospitals [90%], nurses [70%]). Hospitals almost universally offered hepatitis B vaccination to nurses (251/261, 96%), but more than one quarter (132/473, 28%) of nurses reported incomplete vaccination. Provision to physicians was relatively poor (142/261, 54%). The majority of nurses (> 80%) had been vaccinated with bacille Calmette Guerin vaccine, but hospitals reported variable tuberculosis screening practices. Both hospitals and nurses reported low rates (< 30%) of screening and vaccination provision for varicella and measles-mumps-rubella. Two thirds of NSW hospitals (174/261, 67%) provided annual influenza vaccination. CONCLUSIONS: Even though hepatitis B immunization programs were widespread, their effectiveness could be improved by ensuring that vaccination schedules are completed and by targeting physicians. Varicella and measles-mumps-rubella screening and immunization programs are currently lacking. Better strategies are needed to improve the implementation of health care worker protection guidelines in hospitals. PMID- 12734520 TI - Varicella vaccine safety, incidence of breakthrough, and factors associated with breakthrough in Taiwan. AB - BACKGROUND: Varicella vaccine was first available in Taiwan in 1997. The aims of this study were to investigate varicella vaccine safety and occurrence of breakthrough in Taiwan during the first 3 years. The adverse events, incidence of breakthrough, and factors associated with breakthrough were analyzed. METHODS: A personal interview using a structured questionnaire was conducted for the parents of 1248 children less than 12 years old who were vaccinated between 1998 and 2000. Incidence of adverse events and breakthrough were presented and factors associated with breakthrough were estimated by logistic regression. RESULTS: There were 27 (2.16%) breakthrough cases occurring during the maximum follow up period of 31 months, including 22 very mild or mild cases, 3 moderately severe cases, and 2 severe cases. Compared with those who did not have confirmed history of varicella exposure after vaccination, children with such exposure were approximately 28 times as likely to have breakthrough varicella develop (adjusted odds ratio = 27.75, 95% confidence interval: 6.12-125.78, P =.00). There were 91 (7.3%) reported cases of adverse events, including rash, fever, and pain or swelling, occurring within 2 weeks of vaccination. CONCLUSIONS: Although rare adverse events cannot be well-quantified in this study, the results suggest that, at least in the short term, varicella vaccine is well-tolerated and effective in Taiwan. Long-term monitoring program is necessary to ensure the safety of this vaccine. PMID- 12734522 TI - Commentary: The evolving role of nurses in terrorism and war. AB - The health care needs of people in both developed and underdeveloped countries experiencing violent conflict are strikingly similar. Nurses are key for infectious disease detection and control, social support, and rehabilitation during terrorism and war in many countries. Nurses already provide much of the care in these situations because they are present on a day-to-day basis, have key clinical and organizational skills, and tend to enjoy a high level of popular trust. There is a great potential for the profession to do more with systems training, skill development, and participation in policy and research related to reducing the effects of conflict. This is applicable for both terrorist events in developed countries and for humanitarian crises in developing countries. However, nurses have frequently been invisible, serving without discipline-specific orientation. This is partly the result of unresolved ethical and political issues among nursing leaders regarding the image and role of nursing, humanist values, and relations between the profession and the government. Major nursing organizations frequently support national pro-war policies, whereas nursing leaders more often stress support for an international humanitarian orientation. These issues have been with organized nursing since its origins with Florence Nightingale. With better organization, autonomy, and recognition of nursing's unique contributions, nursing can achieve much more in situations of conflict to prevent harm and facilitate recovery among individuals, families, and their communities. Elements of an agenda for nursing research and development are detailed. PMID- 12734521 TI - Seroprevalence survey of measles, rubella, varicella, and mumps antibodies in health care workers and evaluation of a vaccination program in a tertiary care hospital in Japan. AB - Vaccine-preventable viral infections in health care workers (HCWs) have been on the rise for the past 10 years in Japan. To reduce the viral infections and the burden of exposure follow-up surveys at a tertiary care hospital in Osaka, Japan, a seroprevalence survey was conducted, and free vaccinations for measles, rubella, varicella, and mumps were offered to newly hired HCWs (199 physicians and 72 nurses and nursing assistants) who had negative serologic results for antibodies against these viruses. Negative antibody titers were obtained from 7.4% of the newly hired HCWs for measles, 12.5% for rubella, 4.1% for varicella, and 15.9% for mumps. The vaccination program for HCWs improved the vaccine preventable infection rates and resulted in fewer exposure follow-up surveys, fewer lost work days, and fewer HCWs requiring hospitalization for these viral infections compared with those counted for the previous year. These data indicate that all HCWs should be strongly recommended to be vaccinated against (or have documented immunity to) these viruses in Japan, as is the case in the United States. PMID- 12734523 TI - Transmission of HIV and hepatitis C virus from a nursing home patient to a health care worker. AB - BACKGROUND: We report a case of simultaneous HIV and hepatitis C virus (HCV) transmission from a nursing home patient to a health care worker (HCW) whose HIV and HCV infections were diagnosed during routine blood donor screening. METHODS: Detailed information about the HCW, possible occupational and nonoccupational blood and body fluid exposures, and possible source patient was collected. Blood samples were drawn from the HCW and patient, and HIV and HCV laboratory testing was performed at the Centers for Disease Control and Prevention. RESULTS: The HCW, who worked as a nursing home aide, had no nonoccupational risk factors for HIV or HCV infection but provided care for 1 HIV-infected patient with dementia and urinary and fecal incontinence. The HCW had numerous exposures to the patient's emesis, feces, and urine to unprotected chapped and abraded hands. HCW and patient blood samples were positive for anti-HCV by enzyme immunoassay and recombinant immunoblot assay testing. The HCW's and patient's HCV were genotyped as 1a, and their HIV-1 was genotyped as subtype B. HIV and HCV ribonucleic acid (RNA) sequence analysis showed that the HCW's and patient's viruses were very closely related. CONCLUSIONS: HIV and HCV transmission from the patient to the HCW appears to have occurred through nonintact skin exposure. Bloodborne pathogen transmission may have been prevented in this situation by consistent, unfailing use of barrier precautions. PMID- 12734524 TI - The anthrax team: a novel teaching approach to increase anthrax and bioterrorism awareness. AB - A team approach to educating staff regarding anthrax and bioterrorism awareness was implemented after the acts of terrorism that began September 11, 2001. "The Anthrax Team" developed algorithms on the basis of 5 different scenarios, an educational brochure, and a PowerPoint presentation. With use of the algorithms and PowerPoint material, the team conducted informal educational sessions to increase awareness and allay fears. On the basis of the success of this educational method, the teaching process will be applied to address staff on other biologic and chemical terrorism agents. PMID- 12734525 TI - Anthrax and the mail: the making of an educational video for mail workers. AB - The anthrax bioterrorist attacks in 2001 affected millions of people who process, sort, and deliver mail. To more effectively communicate information intended to protect the health of these workers, the Centers for Disease Control and Prevention produced a short-format educational video in December 2001 that targets this diverse group. This report illustrates how an educational video can be rapidly produced to translate and disseminate public health recommendations as part of a public health emergency response. PMID- 12734526 TI - Use of audit tools to evaluate the efficacy of cleaning systems in hospitals. AB - BACKGROUND: Recent publicity has highlighted both the inadequacies of hospital cleaning and high levels of methicillin-resistant Staphylococcus aureus infections in UK hospitals. "Standards for Environmental Cleanliness" (SEC) was a checklist developed in April 1999 by the Infection Control Nurses Association and the Association of Domestic Managers to evaluate cleaning services regardless of who is the provider. More recently, the National Health Service plan (July 2000) was an attempt to generate a rapid improvement in the cleanliness and tidiness of hospitals via a National Health Service patient environment audit (PEA). On the basis of models used in the food industry to manage cleaning practices cost effectively, a risk-based audit checklist incorporating rapid hygiene monitoring was developed to assess the adequacy of cleaning programs and standards in hospitals. This checklist (Audit for Cleaning Efficacy, or ACE) as well as the SEC and PEA approaches were applied at 4 hospitals, and environmental microbial surface counts were compared. SEC and PEA rely on visual assessment, whereas the ACE approach is more comprehensive and included more specific questions relating to the management and monitoring of cleaning as well as standards on the basis of rapid hygiene monitoring. METHODS: Two wards in each of the 4 hospitals were visited on 3 separate occasions immediately after cleaning was completed. Visual assessment, adenosine triphosphate bioluminescence, and microbiologic sampling of selected environmental sites were performed to evaluate the effectiveness of cleaning. The 3 audits were completed during the final hospital visit. RESULTS: Visual assessment indicated that 90% of sites were satisfactory, whereas adenosine triphosphate bioluminescence showed that 100% and microbiologic sampling showed that 90% of sites did not meet benchmark values. There was no significant difference between the SEC and PEA audits (P =.311), which used visual assessment, and the results suggest that they both are similar in passing surfaces that have microbiologic benchmark values that are too high. However, the ACE audit showed a significant difference (P = <.001) in results compared with the SEC and the PEA audits and did not pass surfaces with microbiologic benchmark values that were too high. The ACE audit, which incorporates rapid hygiene testing, showed a much stronger association with the microbial counts; this was not apparent with the SEC and the PEA audits. CONCLUSION: The data suggest that visual assessment is a poor indicator of cleaning efficacy and that the ACE audit gives a better assessment of cleaning programs compared with the other 2 audit methods in relation to microbial surface counts. It is recommended that hospital cleaning regimes be designed to ensure that surfaces are cleaned adequately and that efficacy is assessed with use of internal auditing and rapid hygiene testing. PMID- 12734527 TI - Effective medical waste management: it can be done. AB - BACKGROUND: It was noticed that a large volume of medical waste was being generated for incineration at our hospital. The 2 incinerators at our facility were unable to effectively deal with the load of waste and, therefore, were operating for extended periods of time. This caused a significant amount of soot and other emissions to be produced as pollutants into the surrounding environment, which is considered to be a real health hazard. METHODS: A waste management plan was introduced that included education, mandatory inservice training, auditing of the type and volume of waste generated by each department, and introduction of a written policy on waste management. RESULTS: Within a few months of implementation of the waste-management plan, the amount of medical waste was reduced by more than 58%, from 609 skips/mo (2000 kg/day) in the year 1999, to 256 skips/mo (850 kg/day) in the year 2000; skips are steel containers filled with infectious waste. This reduction was maintained throughout the year 2001 and lead to a 50% reduction in total financial costs (17,936 US dollars) with savings in fuel of 5262 US dollars, labor-cost savings of 8990 US dollars, and maintenance and spare parts savings of 3680 US dollars. CONCLUSIONS: This article discusses problems encountered in waste management in our health care facility, solutions and control measures introduced, and achievements. It also demonstrates that effective waste management can reduce health risk, save money, and protect the environment. PMID- 12734529 TI - Transient myocardial ischemia is an independent predictor of adverse in-hospital outcomes in patients with acute coronary syndromes treated in the telemetry unit. AB - OBJECTIVE: The purpose of this study was to determine whether transient myocardial ischemia (TMI) is predictive of adverse in-hospital outcomes among patients admitted to a telemetry unit with acute coronary syndrome (ACS). DESIGN AND SETTING: The study was designed as a prospective clinical trial in a telemetry unit of a large, urban, university medical center. SAMPLE: The sample was comprised of adult patients admitted to the telemetry unit for treatment of acute myocardial infarction, unstable angina, or coronary artery disease warranting cardiac catheterization or percutaneous coronary intervention. METHODS: Continuous 12-lead electrocardiographic (ECG) ST-segment monitoring was initiated in patients admitted to the telemetry unit. TMI was defined as a change in ST amplitude of > or = 100 microV (elevation or depression) in > or = 1 ECG lead lasting > or = 60 seconds, comparing a baseline 12-lead ECG with an event ECG. Frequencies, measurements of central tendency, t test, chi(2) test, and logistic regression analysis were used for data analysis. A P value of <.05 was adopted as the critical value to determine statistical significance. RESULTS: In 237 telemetry unit patients, 39 patients (17%) had ischemia. Overall, 46% of the patients with ischemia had in-hospital complications compared with 10% of the group without ischemia (P <.001). After controlling for other predictors of adverse outcomes (eg, age, gender, Norris prognostic indicator), patients with TMI were 8.5 times more likely to have in-hospital complications (95% CI, 3.71 to 19.71). CONCLUSION: TMI is an independent predictor of in-hospital complications among patients with ACS treated in the telemetry unit setting. Continuous 12-lead ECG ST-segment monitoring provides prognostic information for risk stratification of patients admitted to the hospital for treatment of ACS. PMID- 12734530 TI - Effects of a telephone counseling intervention on psychosocial adjustment in women following a cardiac event. AB - OBJECTIVE: The purpose of this study was to test the effect of a post-discharge telephone counseling intervention on women's psychosocial adjustment following a cardiac event. DESIGN: The study was a prospective, randomized, controlled trial. PATIENTS: Women (n = 196) were recruited from 4 hospitals in Sydney, Australia, who were hospitalized for coronary artery disease: myocardial infarction, coronary artery bypass grafts, coronary angioplasty, or stable angina. Women were randomized to usual care (n = 103) or telephone counseling (n = 93) and were 67 years of age (range 34-92). The majority had not completed high school (92%) and were not employed (84%). OUTCOMES: Psychosocial adjustment was measured by the Psychosocial Adjustment to Illness Scale and the Hospital Anxiety and Depression Scale the day before hospital discharge and 12 weeks postdischarge. INTERVENTION: Individualized information and support, was designed to promote self-managed recovery and psychosocial adjustment, and began with an evaluation during admission and was followed up by telephone counseling at 1, 2, 3, and 6 weeks after discharge. RESULTS: The intervention had no effect on psychosocial adjustment (F[1,182] = 0.06, P =.8), anxiety (F[1,182] = 0.15, P =.69) or depression (F[1,182] = 0.11, P =.74) at 12 weeks after discharge. Women made significant improvements during the 12 weeks on mean scores for psychosocial adjustment (F[1,182] = 58.37, P =.00), anxiety (F [1,182] = 74.58, P =.00) and depression (F[1,182] = 14.11, P =.00). The predictors of poor psychosocial outcomes for women included being less than 55 years of age, being unemployed or retired, having poor psychosocial adjustment to illness at baseline, having readmission, or experiencing a stressful, personal event during follow-up. CONCLUSIONS: Women at risk for poor outcomes following hospitalization for a cardiac event can be identified (ie, women less than 55 years of age, unemployed or retired, poorly adjusted to their cardiac illness, or readmitted to hospital within 12 weeks of a previous cardiac admission), but an effective intervention to enhance psychosocial outcomes remains to be established. PMID- 12734531 TI - Identification of factors predictive of hospital readmissions for patients with heart failure. AB - OBJECTIVE: The objective was to evaluate whether severity of cardiac illness, cognitive functioning, and functional health of older adults with heart failure (HF) and psychosocial factors related to caregiving are predictive of hospital readmissions for those with HF. DESIGN: A prospective, descriptive, predictive design was used. SETTING: The study took place in 2 community hospitals in northeastern Ohio. SAMPLE: Originally 156 patient-caregiver dyads were interviewed within 7 to 10 days of hospital discharge, but only 128 dyads completed the study. Subjects had HF and their mean age was 77.3 years. Their caregivers were mostly women with a mean age of 64.8 years. RESULTS: Fourty-four percent of the patients were readmitted to the hospital within 3 months. Among patients, severity of illness was moderate, blood pressure was within normal limits, functional and cognitive status were high. For patients, the interaction of severity of cardiac illness and functional status predicted risk of hospital readmission. Among caregivers, depressive symptoms and perceived stress were low; informal social support and caregiving appraisal were high. The interaction of caregiver stress and depression were significant predictors of risk of hospital readmission. CONCLUSION: Nurses should consistently assess changes in patients' cardiac symptoms in addition to their ability to provide self-care. Since patients with HF are at high risk for readmission, further study is needed to determine whether interventions designed to increase spousal support would decrease hospital readmissions. PMID- 12734532 TI - Case report: Dexatrim (Phenylpropanolamine) as a cause of myocardial infarction. AB - Phenylpropanolamine (PPA) is a sympathetic amine used in over-the-counter cold remedies and weight-control preparations worldwide. Its use has been associated with hypertensive episodes and hemorrhagic strokes in younger women. Several reports have linked the abuse of PPA with myocardial injury, especially when overdose is involved. We report here the first case of Dexatrim (PPA)-induced myocardial injury in a young woman who was using it at recommended doses for weight control. In addition, we review the 7 other cases of PPA related myocardial injury that have been reported so far. Physicians and patients should be alert to the potential cardiac risk associated with the use of PPA, even at doses generally considered to be safe. PMID- 12734533 TI - Depression and anxiety in adults with congenital heart disease: a pilot study. AB - OBJECTIVE: To examine the presence of depression and anxiety in adults with congenital heart disease and the association of medical severity with depression and anxiety. DESIGN: Prospective, pilot study. SETTING: An adult outpatient cardiology clinic in university-affiliated children's hospital in Northeast. PATIENTS: Twenty-two adult patients with congenital heart disease followed in an adult cardiology clinic. Patients were selected who had no evidence of emotional or behavioral difficulties, (ie, no symptoms of depression or anxiety). Outcome Measures Standardized semi-structured psychiatric interview with structured checklist eliciting Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) diagnostic criteria for depressive and anxiety disorders, Brief Symptom Inventory (BSI), and the Cardiologist's Perception of Medical Severity scale. RESULTS: Among patients who had been assumed to be "well adjusted," 36.4% were experiencing a diagnosable psychiatric disorder, with anxiety or depressive symptoms being prominent. There were 6 patients (27.3%) who had BSI scores above 63 indicating pathological emotional functioning. There was significant convergent validity between the clinical diagnoses of depression and anxiety using both diagnostic interviews to identify DSM-IV diagnostic conditions and the BSI. There were significant associations between patient's medical severity scores and with the DSM-IV diagnosis of depression and the BSI global index score and depression subscale. CONCLUSIONS: This pilot study raises important concerns about the emotional functioning of many adults facing congenital heart disease, particularly those with complex lesions. From a clinical perspective, this work suggests that health care professionals should be alert for emotional difficulties and the possible need for psychological intervention for adult survivors of congenital heart disease even among those that are seemingly emotionally well adjusted. PMID- 12734535 TI - Comparison of oxygen consumption in performing daily activities between patients with chronic obstructive pulmonary disease and a healthy population. AB - OBJECTIVE: The purposes of this study were to compare the daily activity oxygen consumption (VO(2)) and peak oxygen consumption (VO(2peak)) for chronic obstructive pulmonary disease (COPD) patients and healthy individuals; to compare dyspnea levels found in COPD patients and healthy individuals when they performed daily activities and exercise tests; and to establish standard VO(2) values for daily activities for COPD patients. DESIGN: This was an exploratory and correlative study. SETTING: The study took place at the Research Center of Sports Medicine at Taipei Medical University, in Taipei, Taiwan. SUBJECTS: The study included 27 COPD patients and 18 healthy subjects whose ages, weights, and heights were matched. OUTCOME MEASURES: VO(2peak) and the VO(2) for performing daily activities including sitting, standing, walking, walking with a 2-kg load, and walking upstairs for 2 stories. INTERVENTION: All data were collected by means of questionnaires and treadmill exercise tests. VO(2) was measured using an AEROSPORT KB1-C metabolic measurement system. RESULTS: There was no significant difference in VO(2) found between the 2 groups when they were performing daily activities, but the VO(2peak) was significantly lower in the COPD group (13.90 +/ 2.93 mL kg(-1) min(-1)) compared with the healthy control group (16.15 +/- 1.86 mL kg(-1) min(-1)) (P =.01). The dyspnea level of the COPD group when they were performing daily activities and exercise tests was more severe than that of the healthy control group. The mean VO(2) values for daily activities in COPD patients were as follows: sitting 3.41 (+/-0.82), standing 3.67 (+/-0.90), walking 10.06 (+/-2.19), walking with a 2-kg load 10.28, and walking upstairs 8.16 (+/-1.36) mL kg(-1) min(-1). CONCLUSION: The results of this study reveal that there were no differences in VO(2) values for performing daily activities between COPD patients and healthy individuals. However, an increase in dyspnea level occurred during daily activities, and it was found to be more severe for COPD patients than for healthy individuals. A key factor was probably that COPD patients had an obviously lower VO(2peak) and higher relative exercise intensity for daily activities than did healthy individuals. PMID- 12734534 TI - The effect of dobutamine infusion on fractional diaphragm thickening and diaphragm blood flow during fatigue. AB - BACKGROUND: Diaphragm fatigue (DF) has been implicated in respiratory failure in diseases that increase inspiratory resistance loading (IRL) and may complicate weaning of patients from mechanical ventilation. OBJECTIVE: The purpose of this study was to examine the effects of dobutamine administration (10 micro g/kg/min) on DF and to identify the mechanisms by which dobutamine augments diaphragm shortening and diaphragm blood flow (DBF) during fatigue with a rat model. METHODS: The study had an experimental design with 3 groups of Sprague-Dawley rats (n = 38) with 4 experimental periods: period 1, control; period 2, application of IRL; period 3, treatment; and period 4, recovery. DF was produced via IRL. During period 3 treatment, normal saline solution was infused in group I, dobutamine in group II, and dobutamine plus butoxamine hydrochloride in group III. The percent change in fractional diaphragm thickness (FDT) during inspiration reflected diaphragm shortening. DBF and aortic blood flow were determined with fluorescent microspheres. Diaphragm vascular resistance and systemic vascular resistance were calculated on the basis of Poiseuille's equation. RESULTS: Results indicated infusion of dobutamine increased FDT (P =.01) and DBF (P =.009) with respect to fatigue levels. The effects of dobutamine on FDT and DBF were attenuated with infusion of butoxamine hydrochloride (a beta 2 adrenoceptor antagonist) with respect to fatigue. CONCLUSION: Administration of dobutamine at a rate similar to that used clinically increased diaphragm muscle contractility (FDT) and DBF in diaphragms fatigued by IRL. The dobutamine effect on FDT may be the result of restoration of the balance between diaphragm energy consumption and energy delivery to the diaphragm by increasing DBF. Butoxamine hydrochloride attenuated the dobutamine-induced increase in DBF, indicating dobutamine produced vasodilatation via beta-2 adrenoceptors. Thus, the administration of intravenous dobutamine may be a useful adjunct in the treatment of DF. PMID- 12734536 TI - Health perceptions and risk behaviors of lung cancer survivors. AB - BACKGROUND: Lung cancer survivors are at an increased risk for recurrence and the development of secondary tumors and other comorbid conditions. However, little is known about lung cancer survivors' risk behaviors and the effect of these behaviors on overall health perceptions. OBJECTIVE: The purpose of this study was to describe the prevalence of health risk behaviors among non-small cell lung cancer survivors and their relationship perception to overall health. METHODS: One hundred forty-two survivors of non-small cell lung cancer with a minimum of 5 years disease free completed a battery of questionnaires to assess perception of health status and self-reported risk behaviors (smoking, exposure to secondhand smoke, alcohol use) and weight before and after diagnosis. Urinary cotinine level was used to verify smoking status, and actual height and weight were obtained to determine overweight status (body mass index, >/=25). Descriptive statistics and logistic regression were used to analyze the data. RESULTS: Seventy percent of participants reported their health as good to excellent. Although 81% quit smoking after diagnosis, 13.4% continued to smoke and 28% reported exposure to secondhand smoke. Approximately half the sample (58%) drank alcohol (16.3% quit after diagnosis) and was overweight (51%). A strong agreement between current smoking and exposure to secondhand smoke was observed. In a multivariate analysis, smoking (odds ratio [OR], 7.02; CI, 2.45 to 20.13), exposure to secondhand smoke (OR, 5.37; CI, 2.42 to 11.95), alcohol use (OR, 9.04; CI, 3.28 to 24.92), and overweight (OR, 8.51; CI, 3.44 to 21.10) were independent predictors of perceived poor health status. CONCLUSION: Although most lung cancer survivors have made healthy lifestyle changes, a substantial proportion has not. Our findings suggest the need for multiple risk factor interventions to decrease risk behaviors and improve overall health after a cancer diagnosis. PMID- 12734537 TI - Streptococcal viridans subacute bacterial endocarditis associated with antineutrophil cytoplasmic autoantibodies (ANCA). AB - We report an illustrative case of a 60-year-old man with Streptococcus viridans subacute bacterial endocarditis (SBE) and positive antineutrophil cytoplasmic autoantibodies (c-ANCA). C-ANCA positivity has been associated with a variety of rheumatic and infectious disease areas, but has been rarely associated with SBE. The patient had mitral valve prolapse with mitral regurgitation, and S viridans SBE developed after a dental procedure. Laboratory abnormalities included anemia, elevated erythrocyte sedimentation rate, positive rheumatoid factor, positive anticardiolipin antibody, positive lupus anticoagulant, and highly elevated c ANCA level. We believe this is only the ninth reported case of S viridans SBE with a positive c-ANCA, and the third with mitral valve prolapse and vegetations. PMID- 12734539 TI - PCR-generated padlock probes distinguish homologous chromosomes through quantitative fluorescence analysis. AB - Conventional cytogenetic techniques can distinguish homologous chromosomes in a qualitative manner based upon obvious morphological features or using in situ hybridization methods that yield qualitative data. We have developed a method for quantitative genotyping of single-nucleotide variants in situ using circularizable DNA probes, so-called padlock probes, targeting two different alpha satellite repeat variants present in human chromosome 7 centromeres, and a single-nucleotide variation in alpha satellite repeats on human chromosome 15 centromeres. By using these PCR-generated padlock probes, we could quantitatively distinguish homologous chromosomes and follow the transmission of the chromosomes by in situ analysis during three consecutive generations. PMID- 12734538 TI - Single lead dual-chamber pacing. PMID- 12734540 TI - Cys611Ser mutation in RET proto-oncogene in a kindred with medullary thyroid carcinoma and Hirschsprung's disease. AB - Germline mutations in the RET proto-oncogene are responsible for the development of human hereditary diseases, including multiple endocrine neoplasia (MEN) type 2A and 2B, familial medullary thyroid carcinoma (FMTC), and Hirschsprung's disease (HSCR). It has been reported that some families developed both MEN 2A/FMTC and HSCR, in which a mutation in a cysteine residue at codon 609, 618, or 620 in the RET gene was present. Here we report a novel RET mutation detected in a Japanese family with medullary thyroid carcinoma and HSCR. A germline mutation in cysteine 611 of the RET gene was identified in this family, which introduced an amino-acid change from cysteine to serine. By biological and biochemical analyses of mutant RET proteins, we previously predicted the potentiality that amino-acid substitution for cysteine 611 as well as cysteines 609, 618, and 620 would promote the development of MEN 2A/FMTC and HSCR. This clinical case substantiates our suggestion for the mechanism of the development of both the diseases. PMID- 12734541 TI - Transient expression of wild-type and mutant glucocerebrosidases in hybrid vaccinia expression system. AB - Gaucher disease, the most prevalent lysosomal storage disease, is characterised by a significant phenotypic variation caused by more than 150 mutations. In order to verify pathogenicity of mutations found in the Czech Gaucher population, the vaccinia expression system was used. The wild-type human beta-glucocerebrosidase cDNA and cDNAs carrying the mutations 72delC, 1326insT, 1263del55, S196P, N370S, L444P, G202E, D409H, T369M, L444P+V460V, and D409H+T369M were expressed in Gaucher fibroblast cell line (L444P/S107L), BSC40, and HeLa G cells. The enzymatic activity and immunological reactivity were analysed. Only beta glucocerebrosidase-deficient fibroblasts were suitable for expression using plasmid transfection. The expressed beta-glucosidase activity of mutant glucocerebrosidases was in good correlation with the presumed severity of the mutations. PMID- 12734542 TI - Congenital cataract as the first symptom of a neuromuscular disease caused by a novel single large-scale mitochondrial DNA deletion. AB - The male proband reported here was born with appropriate anthropometric parameters at term as the second child of healthy nonconsanguineous parents. His only clinical symptom was bilateral congenital cataracts with strabismus at birth, and both lenses were removed surgically at the age of 8 months. The perinatal and infantile period thereafter was clinically uneventful and his psychomotor development appeared almost normal. At the age of 6 years he was hospitalized for slight muscle weakness, minor ptosis, nystagmus and decreased physical activity. Soon after, his general condition worsened, gait ataxia presented, dysphagia and difficulty of speech followed by rapidly progressive generalized ataxia, and myopathy developed. Typical progressive gray matter degeneration with focal necrosis in the basal ganglia characteristic of the Leigh type of neuropathology could be detected by cranial MRI, the muscle histology showed ragged-red fibers. At the age of 7.5 years, unexpected left side hemiparesis with speech disability resembling that seen in MELAS syndrome developed, from which he recovered within 1.5 days. The mtDNA of the patient showed single 6.7 kb large-scale deletion harboring between 7817 and 14 536 bp. This case represents the first report of a verified mtDNA mutation associated with congenital cataracts as the first clinical sign of a later developing progressive neuromuscular disease presented with a combination of Leigh neuropathology, ragged-red fiber histopathology and stroke-like attack. PMID- 12734543 TI - MUC1 polymorphism confers increased risk for intestinal metaplasia in a Colombian population with chronic gastritis. AB - Gastric cancer (GC) stands as the second most common cause of cancer death for males worldwide, and intestinal metaplasia (IM) is a lesion that precedes GC development. In previous works it was shown that polymorphisms of MUC1 gene are associated with increased risk for GC and IM. The aim of the present study was to evaluate MUC1 gene polymorphism in patients with chronic gastritis from Colombia. A Portuguese population of patients with chronic gastritis was used for comparative purposes. A total of 67 Colombian cases and 52 Portuguese cases were analysed by restriction analysis and Southern blotting. MUC1 allele frequencies were significantly different between the two populations, with an overall prevalence of smaller alleles in Colombian samples. Colombian cases showed a lower prevalence of individuals homozygous for small MUC1 mucins in cases without IM (62.5%) when compared with cases with IM (86.0%). The same trend, although not statistically significant, is observed in the Portuguese population. In conclusion, our study shows that Colombian patients with chronic gastritis have a significantly higher prevalence of small MUC1 alleles than the Portuguese population. Our study also shows that small MUC1 genotypes are associated with increased risk for IM development in Colombian patients. PMID- 12734544 TI - Spatial patterns of cystic fibrosis mutation spectra in European populations. AB - Cystic fibrosis (CF) is the most frequent severe recessive disorder in European populations. We have analyzed its mutation frequency spectrum in 94 European, North African and SW Asian populations taken from the literature. Most major mutations as well as the incidence of CF mutations showed clinals patterns as demonstrated by autocorrelogram analysis. More importantly, measures of mutation diversity did also show clinal patterns, with mutation spectra being more diverse in southern than in northern Europe. This increased diversity would imply roughly a three-fold long-term effective population size in southern than in northern Europe. Distances were computed among populations based on their CF mutation frequencies and compared with distances based on other genic regions. CF-based distances correlated with mtDNA but not with Y-chromosome-based distances, which may be a consequence of the relatively homogeneous CF mutation frequencies in European populations. PMID- 12734545 TI - Measured haplotype analysis of the aldosterone synthase gene and heart size. AB - Gene-association studies of heart size and the aldosterone synthase (CYP11B2) gene have produced inconsistent results, possibly because of limitations in the sample size and/or the number and location of the polymorphisms. An analysis of six polymorphisms spanning 6 kb of the CYP11B2 gene in Caucasian British families revealed a limited number of haplotypes because of strong linkage disequilibrium over this small region. The genotype and haplotype information was used in an association study involving 955 members of 229 families phenotyped for echocardiographic measures of heart size. In a mixed effects linear modelling analysis, the G5937C polymorphism was associated with cardiac wall thickness (P=0.02), and the intron conversion and A4550C polymorphisms were associated with left ventricular cavity size (P=0.02 and 0.002, respectively). Measured haplotype analyses confirmed the association of alleles at the intron conversion and G5937C polymorphisms with cardiac wall thickness (P=0.02), and alleles at the intron conversion polymorphism with left ventricular cavity size (P=0.04). The polymorphisms contributed to 2.0-3.4% of the variability in these traits. In summary, genetic polymorphisms at the CYP11B2 gene make a small contribution to quantitative variation in echocardiographic measures of heart size. These results point to the importance of analysing the full extent of genetic variation that captures the haplotype structure of a locus in gene association studies. PMID- 12734547 TI - A new locus for postaxial polydactyly type A/B on chromosome 7q21-q34. AB - Postaxial polydactyly (PAP) is the occurrence of one or more extra ulnar or fibular digits or parts of it. In PAP-A, the extra digit is fully developed and articulates with the fifth or an additional metacarpal/metatarsal, while it is rudimentary in PAP-B. Isolated PAP usually segregates as an autosomal dominant trait, with variable expression. Three loci are known for PAP in humans. PAPA1 (including PAP-A/B in one patient) on 7p13 caused by mutations in the GLI3 gene, PAPA2 on 13q21-q32 in a Turkish kindred with PAP-A only, and a third one (PAPA3) in a Chinese family with PAP-A/B on 19p13.1-13.2. We identified a fourth locus in a large Dutch six-generation family with 31 individuals including 11 affecteds. Their phenotype varied from either PAP-A, or PAP-B to PAP-A/B with or without the co-occurence of partial cutaneous syndactyly. We performed a whole-genome search and found linkage between PAP and markers on chromosome 7q. The highest LOD score was 3.34 obtained at D7S1799 and D7S500 with multipoint analysis. PMID- 12734546 TI - Influence of missense mutation and silent mutation of LHbeta-subunit gene in Japanese patients with ovulatory disorders. AB - The frequency of variant LHbeta containing two point mutations (T(986)-C and T(1008)-C) and its relationship to reproductive disorders differ widely between ethnic groups. In a Japanese population, variant luteinizing hormone (LH) correlates with ovulatory disorders. Here we examined the relationship between two missense mutations and five silent mutations (C(894)-T, G(1018)-C, C(1036)-A, C(1098)-T and C(1423)-T) in the LHbeta gene, and ovulatory disorders. We studied 43 patients with ovulatory disorders, 79 patients with normal ovulatory cycles, and 23 healthy men who agreed to join our DNA analysis. PCR-amplified LHbeta subunit gene sequences were compared with a base sequence of wild-type LH reported after direct sequencing. The highest frequency (0.945) of novel allele was observed at the position of the C(1036)-A transition. No homozygotes for wild type LHbeta (C(1036)) were identified. The frequency of novel allele in patients with polycystic ovary syndrome, endometriosis, premature ovarian failure and luteal insufficiency was significantly different from that of healthy women. The frequencies of novel alleles (C(894)-T, C(1098)-T and C(1423)-T) in patients with ovulatory disorders were significantly higher than those with normal ovulatory cycles. The mean incidence of point mutation in patients with ovulatory disorders was higher than in those with normal ovulatory cycles. Among patients with variant LH, five silent mutations were identified in 87.5% of patients with ovulatory disorders, whereas only a few silent mutations were identified in patients with normal ovulatory cycles. In a Japanese population, five silent mutations of variant LH could have influenced two missense mutations and/or other unknown missense mutations, causing ovulatory disorders. PMID- 12734548 TI - Multiplex single-tube screening for mutations in the Nijmegen Breakage Syndrome (NBS1) gene in Hodgkin's and non-Hodgkin's lymphoma patients of Slavic origin. AB - Patients with Nijmegen Breakage Syndrome (NBS) have a high risk to develop malignant diseases, most frequently B-cell lymphomas. It has been demonstrated that this chromosomal breakage syndrome results from mutations in the NBS1 gene that cause either a loss of full-length protein expression or expression of a variant protein. A large proportion of the known NBS patients are of Slavic origin who carry a major founder mutation 657del5 in exon 6 of the NBS1 gene. The prevalence of this mutation in Slav populations is reported to be high, possibly contributing to higher cancer risk in these populations. Therefore, if mutations in NBS1 are associated with higher risk of developing lymphoid cancers it would be most likely to be observed in these populations. A multiplex assay for four of the most frequent NBS1 mutations was designed and a series of 119 lymphoma patients from Slavic origin as well as 177 healthy controls were tested. One of the patients was a heterozygote carrier of the ACAAA deletion mutation in exon 6 (1/119). No mutation was observed in the control group, despite the reported high frequency (1/177). The power of this study was 30% to detect a relative risk of 2.0. PMID- 12734549 TI - Identification of a locus (LCA9) for Leber's congenital amaurosis on chromosome 1p36. AB - Leber's congenital amaurosis (LCA) is the most common cause of inherited childhood blindness and is characterised by severe retinal degeneration at or shortly after birth. We have identified a new locus, LCA9, on chromosome 1p36, at which the disease segregates in a single consanguineous Pakistani family. Following a whole genome linkage search, an autozygous region of 10 cM was identified between the markers D1S1612 and D1S228. Multipoint linkage analysis generated a lod score of 4.4, strongly supporting linkage to this region. The critical disease interval contains at least 5.7 Mb of DNA and around 50 distinct genes. One of these, retinoid binding protein 7 (RBP7), was screened for mutations in the family, but none was found. PMID- 12734550 TI - Biological Procedures: An electronic techniques journal serving biochemists and biologists. PMID- 12734551 TI - Immunological assays for chemokine detection in in-vitro culture of CNS cells. AB - Herein we review the various methods currently in use for determining the expression of chemokines by CNS cells in vitro. Chemokine detection assays are used in conjuction with one another to provide a comprehensive, biologically relevant assessment of the chemokines which is necessary for correct data interpretation of a specific observed biological effect. The methods described include bioassays for soluble chemokine receptors, RNA extraction, RT-PCR, Real - time quantitative PCR, gene array analysis, northern blot analysis, Ribonuclease Protection assay, Flow cytometry, ELISPOT, western blot analysis, and ELISA. No single method of analysis meets the criteria for a comprehensive, biologically relevant assessment of the chemokines, therefore more than one assay might be necessary for correct data interpretation, a choice that is based on development of a scientific rationale for the method with emphasis on the reliability and relevance of the method. PMID- 12734552 TI - Methodology for Detecting Trace Amounts of Microchimeric DNA from Peripheral Murine White Blood Cells by Real-Time PCR. AB - Real-time PCR methodology can successfully quantitate microchimeric cell populations at a concentration of 100 microchimeric cells/100,000 host cells; however, it has not been successful in quantitating DNA from trace numbers of microchimeric white blood cells which we reported are present in murine peripheral blood at a concentration as low as 2/100,000 host cells. We report methodology using primers for a portion of the H2-k(b) murine histocompatibility sequence, specific for the C57BL/6J mouse. When these primers were used in the presence of 11,000 microM primer, a 20-fold increase in the median manufacturer's recommended concentration, the assay could be optimized to detect 34 pg of C57BL/6J DNA in a background of 2.5 microg of carrier BALB/cJ DNA (1/100,000). These conditions resulted in a detection limit half as sensitive as that found when no carrier DNA was present. PMID- 12734553 TI - Use of Site-Specifically Tethered Chemical Nucleases to Study Macromolecular Reactions. AB - During a complex macromolecular reaction multiple changes in molecular conformation and interactions with ligands may occur. X-ray crystallography may provide only a limited set of snapshots of these changes. Solution methods can augment such structural information to provide a more complete picture of a macromolecular reaction. We analyzed the changes in protein conformation and protein:nucleic acid interactions which occur during transcription initiation by using a chemical nuclease tethered to cysteines introduced site-specifically into the RNA polymerase of bacteriophage T7 (T7 RNAP). Changes in cleavage patterns as the polymerase steps through transcription reveal a series of structural transitions which mediate transcription initiation. Cleavage by tethered chemical nucleases is seen to be a powerful method for revealing the conformational dynamics of macromolecular reactions, and has certain advantages over cross linking or energy transfer approaches. PMID- 12734554 TI - Application of a Colorimetric Assay to Identify Putative Ribofuranosylaminobenzene 5'-Phosphate Synthase Genes Expressed with Activity in Escherichia coli. AB - Tetrahydromethanopterin (H(4)MPT) is a tetrahydrofolate analog originally discovered in methanogenic archaea, but later found in other archaea and bacteria. The extent to which H(4)MPT occurs among living organisms is unknown. The key enzyme which distinguishes the biosynthetic pathways of H(4)MPT and tetrahydrofolate is ribofuranosylaminobenzene 5'-phosphate synthase (RFAP synthase). Given the importance of RFAP synthase in H(4)MPT biosynthesis, the identification of putative RFAP synthase genes and measurement of RFAP synthase activity would provide an indication of the presence of H(4)MPT in untested microorganisms. Investigation of putative archaeal RFAP synthase genes has been hampered by the tendency of the resulting proteins to form inactive inclusion bodies in Escherichia coli. The current work describes a colorimetric assay for measuring RFAP synthase activity, and two modified procedures for expressing recombinant RFAP synthase genes to produce soluble, active enzyme. By lowering the incubation temperature during expression, RFAP synthase from Archaeoglobus fulgidus was produced in E. coli and purified to homogeneity. The production of active RFAP synthase from Methanothermobacter thermautotrophicus was achieved by coexpression of the gene MTH0830 with a molecular chaperone. This is the first direct biochemical identification of a methanogen gene that codes for an active RFAP synthase. PMID- 12734555 TI - Evaluation of intra- and interspecific divergence of satellite DNA sequences by nucleotide frequency calculation and pairwise sequence comparison. AB - Satellite DNA sequences are known to be highly variable and to have been subjected to concerted evolution that homogenizes member sequences within species. We have analyzed the mode of evolution of satellite DNA sequences in four fishes from the genus Diplodus by calculating the nucleotide frequency of the sequence array and the phylogenetic distances between member sequences. Calculation of nucleotide frequency and pairwise sequence comparison enabled us to characterize the divergence among member sequences in this satellite DNA family. The results suggest that the evolutionary rate of satellite DNA in D. bellottii is about two-fold greater than the average of the other three fishes, and that the sequence homogenization event occurred in D. puntazzo more recently than in the others. The procedures described here are effective to characterize mode of evolution of satellite DNA. PMID- 12734556 TI - A targeted extracellular approach for recording long-term firing patterns of excitable cells: a practical guide. AB - Excitable cells in many endocrine and neuronal systems display rhythms with periodicities on the order of many minutes. To observe firing patterns that represent the output of these rhythms requires a recording technique that can monitor electrophysiological activity for several hours without affecting cell behavior. A targeted extracellular approach (also known as loose-patch) accomplishes this objective. Because low resistance seals (<20 MOmega) do not influence the cell membrane and because the normal intracellular milieu is maintained, this approach is the least invasive method for monitoring the endogenous electrical activity of single cells. In this report, we detail our use of this technique to record the firing patterns of gonadotropin-releasing hormone (GnRH) neurons in brain slices continuously for several hours. PMID- 12734557 TI - Optimization of Naked DNA Delivery for Interferon Subtype Immunotherapy in Cytomegalovirus Infection. AB - Type I interferon (IFN) gene therapy modulates the immune response leading to inflammatory heart disease following cytomegalovirus (CMV) infection in a murine model of post-viral myocarditis. Efficacy of different immunisation protocols for the IFN constructs was influenced by the dose of DNA, subtype choice, combination use, pre-medication, and timing of DNA administration. Optimal efficacy was found with bupivacaine treatment prior to DNA inoculation of 200mg IFN DNA 14 days prior to virus challenge. Maximal antiviral and antimyocarditic effects were achieved with this vaccination schedule. Furthermore, inoculation of synergistic IFN subtypes demonstrated enhanced efficacy when delivered either alone or with CMV gB DNA vaccination in the CMV model. Thus naked DNA delivery of IFN provides an avenue of immunotherapy for regulating herpesvirus-induced diseases. PMID- 12734559 TI - A semi-quantitative RT-PCR method to measure the in vivo effect of dietary conjugated linoleic acid on porcine muscle PPAR gene expression. AB - Conjugated linoleic acid (CLA) can activate (in vitro) the nuclear transcription factors known as the peroxisome proliferators activated receptors (PPAR). CLA was fed at 11 g CLA/kg of feed for 45d to castrated male pigs (barrows) to better understand long term effects of PPAR activation in vivo. The barrows fed CLA had lean muscle increased by 3.5% and overall fat reduced by 9.2% but intramuscular fat (IMF %) was increased by 14% (P < 0.05). To measure the effect of long term feeding of CLA on porcine muscle gene expression, a semi-quantitative RT-PCR method was developed using cDNA normalized against the housekeeping genes cyclophilin and beta-actin. This method does not require radioactivity or expensive PCR instruments with real-time fluorescent detection. PPARgamma and the PPAR responsive gene AFABP but not PPARalpha were significantly increased (P < 0.05) in the CLA fed pig's muscle. PPARalpha and PPARgamma were also quantitatively tested for large differences in gene expression by western blot analysis but no significant difference was detected at this level. Although large differences in gene expression of the PPAR transcriptional factors could not be confirmed by western blotting techniques. The increased expression of AFABP gene, which is responsive to PPAR transcriptional factors, confirmed that dietary CLA can induce a detectable increase in basal PPAR transcriptional activity in the live animal. PMID- 12734558 TI - Progressive Rearrangement of Telomeric Sequences Added to Both the ITR Ends of the Yeast Linear pGKL Plasmid. AB - Relocation into the nucleus of the yeast cytoplasmic linear plasmids was studied using a monitor plasmid pCLU1. In Saccharomyces cerevisiae, the nuclearly relocated pCLU1 replicated in a linear form (termed pTLU-type plasmid) which carried the host telomeric repeats TG(1-3) of 300-350 bp at both ends. The telomere sequences mainly consisted of a major motif TGTGTGGGTGTGG which was complementary to part of the RNA template of yeast telomerase and were directly added to the very end of the pCLU1-terminal element ITR (inverted terminal repeat), suggesting that the ITR end played a role as a substrate of telomerase. The telomere sequences varied among isolated pTLU-type plasmids, but the TG(1-3) organization was symmetrically identical on both ends of any one plasmid. During cell growth under non-selective condition, the telomeric repeat sequences were progressively rearranged on one side, but not on the opposite side of pTLU plasmid ends. This indicates that the mode of telomeric DNA replication or repair differed between both ends. Clonal analysis showed that the intense rearrangement of telomeric DNA was closely associated with extreme instability of pTLU plasmids. PMID- 12734560 TI - Expression, Purification and Characterization of Ricin vectors used for exogenous antigen delivery into the MHC Class I presentation pathway. AB - Disarmed versions of the cytotoxin ricin can deliver fused peptides into target cells leading to MHC class I-restricted antigen presentation [Smith et al. J Immunol 2002; 169:99-107]. The ricin delivery vector must contain an attenuated catalytic domain to prevent target cell death, and the fused peptide epitope must remain intact for delivery and functional loading to MHC class I molecules. Expression in E. coli and purification by cation exchange chromatography of the fusion protein is described. Before used for delivery, the activity of the vector must be characterized in vitro, via an N-glycosidase assay, and in vivo, by a cytotoxicity assay. The presence of an intact epitope must be confirmed using mass spectrometry by comparing the actual mass with the predicted mass. PMID- 12734561 TI - Experimental Assessment of the Role of Acetaldehyde in Alcoholic Cardiomyopathy. AB - Alcoholism is one of the major causes of non-ischemic heart damage. The myopathic state of the heart due to alcohol consumption, namely alcoholic cardiomyopathy, is manifested by cardiac hypertrophy, compromised ventricular contractility and cardiac output. Several mechanisms have been postulated for alcoholic cardiomyopathy including oxidative damage, accumulation of triglycerides, altered fatty acid extraction, decreased myofilament Ca(2+ )sensitivity, and impaired protein synthesis. Despite intensive efforts to unveil the mechanism and ultimate toxin responsible for alcohol-induced cardiac toxicity, neither has been clarified thus far. Primary candidates for the specific toxins are ethanol, its first and major metabolic product - acetaldehyde (ACA) and fatty acid ethyl esters. Evidence from our lab suggests that ACA directly impairs cardiac function and promotes lipid peroxidation resulting in oxidative damage. The ACA-induced cardiac contractile depression may be reconciled with inhibitors of Cytochrome P 450 oxidase, xanthine oxidase and lipid peroxidation Unfortunately, the common methods to investigate the toxicity of ACA have been hampered by the fact that direct intake of ACA is toxic and unsuitable for chronic study, which is unable to provide direct evidence of direct cardiac toxicity for ACA. In order to overcome this obstacle associated with the chemical properties of ACA, our laboratory has used the chronic ethanol feeding model in transgenic mice with cardiac over-expression of alcohol dehydrogenase (ADH) and an in vitro ventricular myocyte culture model. The combination of both in vivo and in vitro approaches allows us to evaluate the role of ACA in ethanol-induced cardiac toxicity and certain cellular signaling pathways leading to alcoholic cardiomyopathy. PMID- 12734562 TI - Aequorin-based measurements of intracellular Ca2+-signatures in plant cells. AB - Due to the involvement of calcium as a main second messenger in the plant signaling pathway, increasing interest has been focused on the calcium signatures supposed to be involved in the patterning of the specific response associated to a given stimulus. In order to follow these signatures we described here the practical approach to use the non-invasive method based on the aequorin technology. Besides reviewing the advantages and disadvantages of this method we report on results showing the usefulness of aequorin to study the calcium response to biotic (elicitors) and abiotic stimuli (osmotic shocks) in various compartments of plant cells such as cytosol and nucleus. PMID- 12734563 TI - A proteomic approach based on peptide affinity chromatography, 2-dimensional electrophoresis and mass spectrometry to identify multiprotein complexes interacting with membrane-bound receptors. AB - There is accumulating evidence that membrane-bound receptors interact with many intracellular proteins. Multiprotein complexes associated with ionotropic receptors have been extensively characterized, but the identification of proteins interacting with G protein-coupled receptors (GPCRs) has so far only been achieved in a piecemeal fashion, focusing on one or two protein species. We describe a method based on peptide affinity chromatography, two-dimensional electrophoresis, mass spectrometry and immunoblotting to identify the components of multiprotein complexes interacting directly or indirectly with intracellular domains of GPCRs or, more generally, any other membrane-bound receptor. Using this global approach, we have characterized multiprotein complexes that bind to the carboxy-terminal tail of the 5-hydroxytryptamine type 2C receptor and are important for its subcellular localization in CNS cells (Becamel et al., EMBO J., 21(10): 2332, 2002). PMID- 12734564 TI - A Sensitive Quantification of HHV-6B by Real-time PCR. AB - Human herpesvirus (HHV)-6B is a pathogen causing latent infection in virtually all humans. Nevertheless, the interaction of HHV-6B with its host cells is poorly understood. Although HHV-6B is approximately 90% homologous to HHV-6A, it expresses certain B-specific genes. In order to quantify the amount of expressed viral mRNA we have developed a method using real-time PCR on a LightCycler instrument. Here we describe an assay for the detection of the HHV-6B B6 mRNA, but our approach can easily be extended to involve other mRNAs. This method is useful during the study of HHV-6B biology and offers reliable and reproducible, quantitative detection of viral mRNA below the attomol range. PMID- 12734565 TI - Use of an Immobilized Monoclonal Antibody to Examine Integrin alpha5beta1 Signaling Independent of Cell Spreading. AB - Cell attachment to the extracellular matrix (ECM) engages integrin signaling into the cell, but part of the signaling response also stem from cell spreading (3). To analyze specific integrin signaling-mediated responses independent of cell spreading, we developed a method engaging integrin signaling by use of an immobilized anti-integrin monoclonal antibody (mab) directed against the fibronectin (FN) receptor integrin alpha5beta1. ECV 304 cells were plated onto FN or immobilized mab JBS5 (anti-integrin alpha5beta1) or onto poly-L-lysin (P-L-L), which mediates integrin-independent attachment. Cells attached and spread on FN, while cells on JBS5 or P-L-L attached but did not spread. Importantly, plating onto FN or mab JBS5 gave rise to identical integrin-induced responses, including a down-regulation of the cyclin-dependent kinase (Cdk2) inhibitors p21(CIP1) and p27(KIP1), while attachment to P-L-L did not. We conclude that engagement of the FN-receptor integrin alpha5beta1 induces integrin signaling regulating the Cdk2 inhibitors independent of cell spreading and present a method for how integrin signaling can be analyzed separate from the effects of cell spreading. PMID- 12734566 TI - PCR-based detection of a rare linear DNA in cell culture. AB - The described method allows for detection of rare linear DNA fragments generated during genomic deletions. The predicted limit of the detection is one DNA molecule per 10(7) or more cells. The method is based on anchor PCR and involves gel separation of the linear DNA fragment and chromosomal DNA before amplification. The detailed chemical structure of the ends of the linear DNA can be defined with the use of additional PCR-based protocols. The method was applied to study the short-lived linear DNA generated during programmed genomic deletions in a ciliate. It can be useful in studies of spontaneous DNA deletions in cell culture or for tracking intracellular modifications at the ends of transfected DNA during gene therapy trials. PMID- 12734568 TI - Primary culture and mRNA analysis of human ovarian cells. AB - Established cell lines are invaluable for studying cell and molecular biological questions. A variety of human ovarian cancer (OC) cell lines exist, however, most have acquired significant genetic alterations from their cells of origin, including deletion of important cell cycle regulatory genes. In order to analyze signaling events related to cell cycle control in human OC, we have modified existing protocols for isolating and culturing OC cells from patient ascites fluid and normal ovarian surface epithelial (OSE) cells from benign ovarian tissue sections. These cells maintain an epithelial phenotype and can be manipulated experimentally for several passages before cellular senescence. An example using TGFb1 treatment of OC cells to examine signaling and target gene activation is presented. PMID- 12734567 TI - A Method for Analyzing the Ubiquitination and Degradation of Aurora-A. AB - The cell cycle machinery consists of regulatory proteins that control the progression through the cell cycle ensuring that DNA replication alternates with DNA segregation in mitosis to maintain cell integrity. Some of these key regulators have to be degraded at each cell cycle to prevent cellular dysfunction. Mitotic exit requires the inactivation of cyclin dependent kinase1 (cdk1) and it is the degradation of the cyclin subunit that inactivates the kinase. Cyclin degradation has been well characterized and it was shown that it is ubiquitin proteasome pathway that leads to the elimination of cyclins. By now, many other regulatory proteins were shown to be degraded by the same pathway, among them members of the aurora kinase family, degraded many other regulatory proteins. Aurora kinases are involved in mitotic spindle formation as well as in cytokinesis. The abundance and activity of the kinase is precisely regulated during the cell cycle. To understand how proteolysis regulates transitions through the cell cycle we describe two assays for ubiquitination and degradation of xenopus aurora kinase A using extracts from xenopus eggs or somatic cell lines. PMID- 12734569 TI - An Efficient Ligation Method in the Making of an in vitro Virus for in vitro Protein Evolution. AB - The "in vitro virus" is a molecular construct to perform evolutionary protein engineering. The "virion (=viral particle)" (mRNA-peptide fusion), is made by bonding a nascent protein with its coding mRNA via puromycin in a test tube for in vitro translation. In this work, the puromycin-linker was attached to mRNA using the Y-ligation, which was a method of two single-strands ligation at the end of a double-stranded stem to make a stem-loop structure. This reaction gave a yield of about 95%. We compared the Y-ligation with two other ligation reactions and showed that the Y-ligation gave the best productivity. An efficient amplification of the in vitro virus with this "viral genome" was demonstrated. PMID- 12734570 TI - Methods for Analysis of Matrix Metalloproteinase Regulation of Neutrophil Endothelial Cell Adhesion. AB - Recent evidence indicates novel role for matrix metalloproteinases (MMPs), in particular gelatinase A (MMP-2), in the regulation of vascular biology that are unrelated to their well-known proteolytic breakdown of matrix proteins. We have previously reported that MMP-2 can modulate vascular reactivity by cleavage of the Gly32-Leu33 bound in big endothelin-1 (ET-1) yielding a novel vasoactive peptide ET-1[1-32]. These studies were conducted to investigate whether gelatinolytic MMPs could affect neutrophil-endothelial cell attachment. ET-1[1 32] produced by MMP-2 up-regulated CD11b/CD18 expression on human neutrophils, thereby promoted their adhesion to cultured endothelial cells. ET-1[1-32] evoked release of gelatinase B (MMP-9), which in turn cleaved big ET-1 to yield ET-1[1 32], thus revealing a self-amplifying loop for ET-1[1-32] generation. ET-1[1-32] was rather resistant to cleavage by neutrophil proteases and further metabolism of ET-1[1-32] was not a prerequisite for its biological actions on neutrophils. The neutrophil responses to ET-1[1-32] were mediated via activation of ET(A)receptors through activation of the Ras/Raf-1/MEK/ERK signaling pathway. These results suggest a novel role for gelatinase A and B in the regulation of neutrophil functions and their interactions with endothelial cells. Here we describe the methods in detail as they relate to our previously published work. PMID- 12734571 TI - High-Yield Method for Isolation and Culture of Endothelial Cells from Rat Coronary Blood Vessels Suitable for Analysis of Intracellular Calcium and Nitric Oxide Biosynthetic Pathways. AB - We describe here a method for isolating endothelial cells from rat heart blood vessels by means of coronary microperfusion with collagenase. This methods makes it possible to obtain high amounts of endothelial cells in culture which retain the functional properties of their in vivo counterparts, including the ability to uptake fluorescently-labeled acetylated low-density lipoproteins and to respond to vasoactive agents by modulating intracellular calcium and by upregulating intrinsic nitric oxide generation. The main advantages of our technique are: (i) good reproducibility, (ii) accurate sterility that can be maintained throughout the isolation procedure and (iii) high yield of pure endothelial cells, mainly due to microperfusion and temperature-controlled incubation with collagenase which allow an optimal distribution of this enzyme within the coronary vascular bed. PMID- 12734572 TI - Mouse Aortic Ring Assay: A New Approach of the Molecular Genetics of Angiogenesis. AB - Angiogenesis, a key step in many physiological and pathological processes, involves proteolysis of the extracellular matrix. To study the role of two enzymatic families, serine-proteases and matrix metalloproteases in angiogenesis, we have adapted to the mouse, the aortic ring assay initially developed in the rat. The use of deficient mice allowed us to demonstrate that PAI-1 is essential for angiogenesis while the absence of an MMP, MMP-11, did not affect vessel sprouting. We report here that this model is attractive to elucidate the cellular and molecular mechanisms of angiogenesis, to identify, characterise or screen "pro- or anti-angiogenic agents that could be used for the treatment of angiogenesis-dependent diseases. Approaches include using recombinant proteins, synthetic molecules and adenovirus-mediated gene transfer. PMID- 12734573 TI - Isolation, growth and identification of colony-forming cells with erythroid, myeloid, dendritic cell and NK-cell potential from human fetal liver. AB - The study of hematopoietic stem cells (HSCs) and the process by which they differentiate into committed progenitors has been hampered by the lack of in vitro clonal assays that can support erythroid, myeloid and lymphoid differentiation. We describe a method for the isolation from human fetal liver of highly purified candidate HSCs and progenitors based on the phenotypes CD38( )CD34(++) and CD38(+)CD34(++), respectively. We also describe a method for the growth of colony-forming cells (CFCs) from these cell populations, under defined culture conditions, that supports the differentiation of erythroid, CD14/CD15(+) myeloid, CD1a(+) dendritic cell and CD56(+) NK cell lineages. Flow cytometric analyses of individual colonies demonstrate that CFCs with erythroid, myeloid and lymphoid potential are distributed among both the CD38(-) and CD38(+) populations of CD34(++) progenitors. PMID- 12734574 TI - Use of an Anaerobic Chamber Environment for the Assay of Endogenous Cellular Protein-Tyrosine Phosphatase Activities. AB - Protein-tyrosine phosphatases (PTPases) have a catalytic cysteine residue whose reduced state is integral to the reaction mechanism. Since exposure to air can artifactually oxidize this highly reactive thiol, PTPase assays have typically used potent reducing agents to reactivate the enzymes present; however, this approach does not allow for the measurement of the endogenous PTPase activity directly isolated from the in vivo cellular environment. Here we provide a method for using an anaerobic chamber to preserve the activity of the total PTPase complement in a tissue lysate or of an immunoprecipitated PTPase homolog to characterize their endogenous activation state. Comparison with a sample treated with biochemical reducing agents allows the determination of the activatable (reducible) fraction of the endogenous PTPase pool. PMID- 12734575 TI - Detection of Progeny Immune Responses after Intravenous Administration of DNA Vaccine to Pregnant Mice. AB - A number of factors influence the development of tolerance, including the nature, concentration and mode of antigen presentation to the immune system, as well as the age of the host. The studies were conducted to determine whether immunizing pregnant mice with liposome-encapsulated DNA vaccines had an effect on the immune status of their offspring. Two different plasmids (encoding antigens from HIV-1 and influenza virus) were administered intravenously to pregnant mice. At 9.5 days post conception with cationic liposomes, injected plasmid was present in the tissues of the fetus, consistent with trans-placental transfer. When the offspring of vaccinated dams were immunized with DNA vaccine, they mounted stronger antigen-specific immune responses than controls and were protected against challenge by homologous influenza virus after vaccination. Moreover, such immune responses were strong in the offspring of mothers injected with DNA plasmid 9.5 days after coitus. These results suggest that DNA vaccinated mothers confer the antigen-specific immunity to their progeny. Here we describe the methods in detail as they relate to our previously published work. PMID- 12734576 TI - Investigation of Neuronal Cell Type-Specific Gene Expression of Ca2+/Calmodulin dependent Protein Kinase II. AB - The promoter activity of the rat Ca(2+)/calmodulin-dependent protein kinase II gene was analyzed using the luciferase reporter gene in neuronal and non-neuronal cell lines. Neuronal cell type-specific promoter activity was found in the 5' flanking region of alpha and beta isoform genes of the kinase. Silencer elements were also found further upstream of promoter regions. A brain-specific protein bound to the DNA sequence of the 5'-flanking region of the gene was found by gel mobility shift analysis in the nuclear extract of the rat brain, including the cerebellum, forebrain, and brainstem, but not in that of non-neuronal tissues, including liver, kidney and spleen. The luciferase expression system and gel shift analysis can be used as an additional and better index by which to monitor gene expression in most cell types. PMID- 12734577 TI - Methods for the study of ionic currents and Ca2+-signals in isolated colonic crypts. AB - Isolated epithelial cells from intestinal mucosae are a suitable object for the study of the regulation of ion transport in the gut. This regulation possesses a great importance for human and veterinary medicine, as diarrheal diseases, which often are caused by an inadequate activation of intestinal anion secretion, are one of the major lethal diseases of children or young animals. The aim of this paper is to describe a method for the isolation of intact colonic crypts, e.g. for the subsequent investigation of the regulation of anion secretion by the intracellular second messenger, Ca(2+) using electrophysiological and imaging techniques. PMID- 12734578 TI - Promoter analysis by saturation mutagenesis. AB - Gene expression and regulation are mediated by DNA sequences, in most instances, directly upstream to the coding sequences by recruiting transcription factors, regulators, and a RNA polymerase in a spatially defined fashion. Few nucleotides within a promoter make contact with the bound proteins. The minimal set of nucleotides that can recruit a protein factor is called a cis-acting element. This article addresses a powerful mutagenesis strategy that can be employed to define cis-acting elements at a molecular level. Technical details including primer design, saturation mutagenesis, construction of promoter libraries, phenotypic analysis, data analysis, and interpretation are discussed. PMID- 12734579 TI - A fluorescence microscopy method for quantifying levels of prostaglandin endoperoxide H synthase-1 and CD-41 in MEG-01 cells. AB - In platelets, PGHS-1-dependant formation of thromboxane A(2) is an important modulator of platelet function and a target for pharmacological inhibition of platelet function by aspirin. Since platelets are a-nucleated cells, we have used the immortalized human megakaryoblastic cell line MEG-01 which can be induced to differentiate into platelet-like structures upon addition of TPA as a model system to study PGHS-1 gene expression. Using a specific antibody to PGHS-1 we have developed a technique utilizing immunofluorescence microscopy and analysis of multiple digital images to monitor PGHS-1 protein levels as MEG-01 cells were induced to differentiate by a single addition of TPA (1.6 x 10(-8) M) over a period of 8 days. The method represents a rapid and economical alternative to flow cytometry. Using this technique we observed that TPA induced adherence of MEG-01 cells, and only the non-adherent TPA-stimulated cells demonstrated compromised viability. The differentiation of MEG-01 cells was evaluated by the expression of the platelet-specific cell surface antigen, CD-41. The latter was expressed in MEG-01 cells at the later stages of differentiation. We demonstrated a good correlation between PGHS-1 levels and the overall level of cellular differentiation of MEG-01 cells. Furthermore, PGHS-1 protein level, which shows a consistent increase over the entire course of differentiation, can be used as an additional and better index by which to monitor megakaryocyte differentiation. PMID- 12734580 TI - Measurement of Cardiac Mechanical Function in Isolated Ventricular Myocytes from Rats and Mice by Computerized Video-Based Imaging. AB - Isolated adult cardiac ventricular myocytes have been a useful model for cardiovascular research for more than 20 years. With the recent advances in cellular physiology and transgenic techniques, direct measurement of isolated ventricular myocyte mechanics is becoming an increasingly important technique in cardiac physiology that provides fundamental information on excitation contraction coupling of the heart, either in drug intervention or pathological states. The goal of this article is to describe the isolation of ventricular myocytes from both rats and mice, and the use of real-time beat-to-beat simultaneous recording of both myocyte contraction and intracellular Ca(2+) transient. PMID- 12734581 TI - Acid Glycohydrolases in Rat Spermatocytes, Spermatids and Spermatozoa: Enzyme Activities, Biosynthesis and Immunolocalization. AB - Mammalian sperm acrosome contains several glycohydrolases thought to aid in the dispersion and digestion of vestments surrounding the egg. In this study, we have used multiple approaches to examine the origin of acrosome-associated glycohdyrdolases. Mixed spermatogenic cells, prepared from rat testis, were separated by unit gravity sedimentation. The purified germ cells (spermatocytes [SP], round spermatids [RS], and elongated/condensed spermatids [E/CS]) contained several glycohydrolase activities. Metabolic labeling in the cell culture, immunoprecipitation, and autoradiographic approaches revealed that beta-D galactosidase was synthesized in SP and RS in 88/90 kDa forms which undergo processing in a cell-specific manner. Immunohistochemical approaches demonstrated that the enzyme was localized in Golgi membranes/vesicles, and lysosome-like structures in SP and RS, and forming/formed acrosome of E/CS. PMID- 12734583 TI - Cytochemical techniques and energy-filtering transmission electron microscopy applied to the study of parasitic protozoa. AB - The study of parasitic protozoa plays a major role in cell biology, biochemistry and molecular biology. Numerous cytochemical techniques have been developed in order to unequivocally identify the nature of subcellular compartments. Enzyme and immuno-cytochemistry allow the detection of, respectively, enzymatic activity products and antigens in particular sites within the cell. Energy-filtering transmission electron microscopy permits the detection of specific elements within such compartments. These approaches are particularly useful for studies employing antimicrobial agents where cellular compartments may be destroyed or remarkably altered and thus hardly identified by standard methods of observation. In this regard cytochemical and spectroscopic techniques provide valuable data allowing the determination of the mechanisms of action of such compounds. PMID- 12734582 TI - Semiquantitative RT-PCR analysis to assess the expression levels of multiple transcripts from the same sample. AB - We describe a semiquantitative RT-PCR protocol optimized in our laboratory to extract RNA from as little as 10,000 cells and to measure the expression levels of several target mRNAs from each sample. This procedure was optimized on the human erythroleukemia cell line TF-1 but was successfully used on primary cells and on different cell lines. We describe the detailed procedure for the analysis of Bcl-2 levels. Aldolase A was used as an internal control to normalize for sample to sample variations in total RNA amounts and for reaction efficiency. As for all quantitative techniques, great care must be taken in all optimization steps: the necessary controls to ensure a rough quantitative (semi-quantitative) analysis are described here, together with an example from a study on the effects of TGF-beta1 in TF-1 cells. PMID- 12734584 TI - A sensitive non-radioactive in situ hybridization method for the detection of chicken IgG gamma-chain mRNA: a technique suitable for detecting of variety of mRNAs in tissue sections. AB - We established a sensitive non-radioactive in situ hybridization (ISH) method for the detection of chicken IgG gamma-chain mRNA in paraffin sections. RNA probes were transcribed in vitro from cloned chicken IgG CH1 nucleotide sequences with SP6/T7 RNA polymerases in the presence of DIG-UTP. These probes were used for hybridization and were immunodetected using anti-DIG antibodies conjugated to horseradish peroxidase. The immunoreactive products were visualized with DAB H(2)O(2). IgG gamma-chain mRNA-expressing cells were localized in both the spleen and oviductal tissues. This method demonstrated an excellent sensitivity since the ISH signal was clear and the background was negligible. We found that in the spleen IgG gamma-chain mRNA-expressing cells were present mainly in the red pulp, whereas in the oviduct they appeared mainly in the mucosal stroma and not in the mucosal epithelium. PMID- 12734585 TI - Post-Electrophoretic Identification of Oxidized Proteins. AB - The oxidative modification of proteins has been shown to play a major role in a number of human diseases. However, the ability to identify specific proteins that are most susceptible to oxidative modifications is difficult. Separation of proteins using polyacrylamide gel electrophoresis (PAGE) offers the analytical potential for the recovery, amino acid sequencing, and identification of thousands of individual proteins from cells and tissues. We have developed a method to allow underivatized proteins to be electroblotted onto PVDF membranes before derivatization and staining. Since both the protein and oxidation proteins are quantifiable, the specific oxidation index of each protein can be determined. The optimal sequence and conditions for the staining process are (a) electrophoresis, (b) electroblotting onto PVDF membranes, (c) derivatization of carbonyls with 2,4-DNP, (d) immunostaining with anti DNP antibody, and (e) protein staining with colloidal gold. PMID- 12734588 TI - The Metabolic Inhibition Model Which Predicts the Intestinal Absorbability and Metabolizability of Drug: Theory and Experiment. AB - The intestinal absorption of analgesic peptides (leucine enkephalin and kyotorphin) and modified peptides in rat were studied. Although these peptides were not absorbed, the absorbability (absorption clearance) of these peptides were increased in the presence of peptidase inhibitors. In order to kinetically analyze these phenomena, we proposed the metabolic inhibition model, which incorporated the metabolic clearance (metabolizability) with the absorption clearance. Metabolic activity was determined with intestinal homogenates. The higher the metabolic clearance was, the lower was the absorption clearance. The relationships between the absorption clearance and the metabolic clearance of the experimental data as well as of the theoretical values were hyperbolic. This model predicted the maximum absorption clearances of cellobiose-coupled leucine enkephalin (0.654 &mgr;l/min/cm) and kyotorphin (0.247 &mgr;l/min/cm). Details of the experimental methods are described. PMID- 12734587 TI - Expression of a prokaryotic P-type ATPase in E. coli Plasma Membranes and Purification by Ni2+-affinity chromatography. AB - In order to characterize the P-type ATPase from Synechocystis 6803 [Geisler (1993) et al. J. Mol. Biol. 234, 1284] and to facilitate its purification, we expressed an N-terminal 6xHis-tagged version of the ATPase in an ATPase deficient E. coli strain. The expressed ATPase was immunodetected as a dominant band of about 97 kDa localized to the E. coli plasma membranes representing about 20-25% of the membrane protein. The purification of the Synecho-cystis 6xHis-ATPase by single-step Ni-affinity chromatography under native and denaturating conditions is described. ATPase activity and the formation of phosphointermediates verify the full function of the enzyme: the ATPase is inhibited by vanadate (IC(50)= 119 &mgr;M) and the formation of phosphorylated enzyme intermediates shown by acidic PAGE depends on calcium, indicating that the Synechocystis P-ATPase functions as a calcium pump. PMID- 12734589 TI - Methods for the Detection of D-Amino-Acid Oxidase. AB - Four methods (an enzyme activity assay, western blotting, RT-PCR, and northern hybridization) to detect the enzyme D-amino-acid oxidase are described. PMID- 12734586 TI - Yeast Two-Hybrid: State of the Art. AB - Genome projects are approaching completion and are saturating sequence databases. This paper discusses the role of the two-hybrid system as a generator of hypotheses. Apart from this rather exhaustive, financially and labour intensive procedure, more refined functional studies can be undertaken. Indeed, by making hybrids of two-hybrid systems, customised approaches can be developed in order to attack specific function-related problems. For example, one could set-up a "differential" screen by combining a forward and a reverse approach in a three hybrid set-up. Another very interesting project is the use of peptide libraries in two-hybrid approaches. This could enable the identification of peptides with very high specificity comparable to "real" antibodies. With the technology available, the only limitation is imagination. PMID- 12734590 TI - Methods for microbial DNA extraction from soil for PCR amplification. AB - Amplification of DNA from soil is often inhibited by co-purified contaminants. A rapid, inexpensive, large-scale DNA extraction method involving minimal purification has been developed that is applicable to various soil types (1). DNA is also suitable for PCR amplification using various DNA targets. DNA was extracted from 100g of soil using direct lysis with glass beads and SDS followed by potassium acetate precipitation, polyethylene glycol precipitation, phenol extraction and isopropanol precipitation. This method was compared to other DNA extraction methods with regard to DNA purity and size. PMID- 12734591 TI - Methods for the Measurement of a Bacterial Enzyme Activity in Cell Lysates and Extracts. AB - The kinetic characteristics and regulation of aspartate carbamoyltransferase activity were studied in lysates and cell extracts of Helicobacter pylori by three diffirent methods. Nuclear magnetic resonance spectroscopy, radioactive tracer analysis, and spectrophotometry were employed in conjunction to identify the properties of the enzyme activity and to validate the results obtained with each assay. NMR spectroscopy was the most direct method to provide proof of ACTase activity; radioactive tracer analysis was the most sensitive technique and a microtitre-based colorimetric assay was the most cost-and time-efficient for large scale analyses. Freeze-thawing was adopted as the preferred method for cell lysis in studying enzyme activity in situ. This study showed the benefits of employing several different complementary methods to investigate bacterial enzyme activity. PMID- 12734592 TI - Dealing with different methods for Kluyveromyces lactis beta-galactosidase purification. AB - Several micro-scale chromatography-based procedures for purification of the beta galactosidase from the yeast Kluyveromyces lactis were assayed. Purified enzyme was suitable to be used as antigen to induce polyclonal antibodies production. Specific staining of non-denaturing PAGE gels with chromogenic substrates allowed the determination of the number of subunits forming the native enzyme. PMID- 12734593 TI - Inferring Deleterious-Mutation Parameters in Natural Daphnia Populations. AB - Deng and Lynch (1, 2) proposed to characterize deleterious genomic mutations from changes in the mean and genetic variance of fitness traits upon selfing in outcrossing populations. Such observations can be readily acquired in cyclical parthenogens. Selfing and life-table experiments were performed for two such Daphnia populations. A significant inbreeding depression and an increase of genetic variance for all traits analyzed were observed. Deng and Lynch's (2) procedures were employed to estimate the genomic mutation rate (U), mean dominance coefficient (), mean selection coefficient (), and scaled genomic mutational variance (). On average, and (^ indicates an estimate) are 0.84, 0.30, 0.14 and 4.6E-4 respectively. For the true values, the and are lower bounds, and and upper bounds. PMID- 12734595 TI - Science through the Internet: Researching, Evaluating and Citing Websites. AB - This article attempts to convey the joys and frustrations of skimming the Internet trying to find relevant information concerning an academic's work as a scientist, a student or an instructor. A brief overview of the Internet and the "do's and don'ts" for the neophyte as well for the more seasoned "navigator" are given. Some guidelines of "what works and what does not" and "what is out there" are provided for the scientist with specific emphasis for biologists, as well as for all others having an interest in science but with little interest in spending countless hours "surfing the net". An extensive but not exhaustive list of related websites is provided. PMID- 12734594 TI - Synthesis by High-Efficiency Liquid-Phase (HELP) Method of Oligonucleotides Conjugated with High-Molecular Weight Polyethylene Glycols (PEGs). AB - The chemical modification of synthetic oligonucleotides has recently been investigated to improve their pharmacological utilization. In addition to chemical alterations of the backbone and of the heterocyclic bases, their conjugation with amphiphylic moieties, such as the polyethylene glycol has been proposed. The large scale production of these molecules as demanded for commercial purposes is hampered by the heterogeneity of the solid-phase processes and by the low reactivity of high-molecular weight PEGs in solution. A new synthetic procedure based on the recently developed liquid-phase method (HELP), has been set up to overcome these limitations. PMID- 12734596 TI - The method of contact angle measurements and estimation of work of adhesion in bioleaching of metals. AB - In this paper, we present our method for the measurement of contact angles on the surface of minerals during the bioleaching process because the standard deviation obtained in our measurements achieved unexpectedly low error. Construction of a goniometer connected with a specially prepared computer program allowed us to repeat measurements several times over a short time course, yielding excellent results.After defining points on the outline of the image of a drop and its baseline as well of the first approximation of the outline of the drop, an iterative process is initiated that is aimed at fitting the model of the drop and baseline. In turn, after defining the medium for which measurements were made, the work of adhesion is determined according to Young-Dupre equation. Calculations were made with the use of two methods named the L-M and L-Q methods. PMID- 12734598 TI - Methods for direct determination of mitomycin C in aqueous solutions and in urine. AB - Stripping voltammetry (SV) is used to quantitatively determine concentrations of the anti-neoplastic drug mitomycin C (MMC) alone and in mixtures with 5 fluorouracil and cisplatin, both of which are used in combined chemotherapy with MMC. If the accumulation is performed at the potentials of MMC reduction (-0.35 V vs. SCE), reduced MMC is strongly adsorbed at the electrode. It is possible to prepare a MMC-modified electrode, which, after a washing step, is transferred to the background electrolyte to determine MMC by voltammetry. This procedure, which is termed transfer stripping voltammetry (TSV), helps to eliminate interferences and can be applied for a direct determination of MMC alone or in mixtures with other drugs in urine. PMID- 12734597 TI - Role of a Transbilayer pH Gradient in the Membrane Fusion Activity of the Influenza Virus Hemagglutinin: Use of the R18 Assay to Monitor Membrane Merging. AB - It had been suggested that influenza virus-mediated membrane fusion might be dependent on a pH gradient across a target membrane. We have designed experiments in which this issue could be addressed. Two populations of liposomes were prepared, both simulating the plasma membrane of target cells, but with the pH of the internal aqueous medium buffered either at pH 7.4 (physiological cytosol pH) or at pH 5.0 (endosomal pH at which influenza virus displays maximal fusion activity). By monitoring fusion using the R18 assay, we found that the internal pH of the target liposomes did not influence membrane merging as mediated by the influenza virus hemagglutinin, thus demonstrating that a transmembrane pH gradient is not required in this fusion process. PMID- 12734601 TI - Yeast Exoglycoproteins Produced Under NaCl-Stress Conditions as Efficient Cryoprotective Agents. AB - Six extracellular yeast glycoproteins were prepared from three yeast species in osmotic equilibrium and unequilibrium environments and used as non-penetrating cryoadditives. Glycoproteins secreted by the strain Dipodascus australiensis into growth medium containing NaCl (8% w/v) were found to be the most effective cryoadditives. It was possible to use these glycoproteins alone (without DMSO as penetrating agent) for the cryoprotection of the studied yeasts. PMID- 12734599 TI - A Method for Assaying Deubiquitinating Enzymes. AB - A general method for the assay of deubiquitinating enzymes was described in detail using (125)I-labeled ubiquitin-fused alphaNH-MHISPPEPESEEEEEHYC (referred to as Ub-PESTc) as a substrate. Since the tyrosine residue in the PESTc portion of the fusion protein was almost exclusively radioiodinated under a mild labeling condition, such as using IODO-BEADS, the enzymes could be assayed directly by simple measurement of the radioactivity released into acid soluble products. Using this assay protocol, we could purify six deubiquitinating enzymes from chick skeletal muscle and yeast and compare their specific activities. Since the extracts of E. coli showed little or no activity against the substrate, the assay protocol should be useful for identification and purification of eukaryotic deubiquitinating enzymes cloned and expressed in the cells. PMID- 12734600 TI - Coupling Optical and Electrical Measurements in Artificial Membranes: Lateral Diffusion of Lipids and Channel Forming Peptides in Planar Bilayers. AB - Planar lipid bilayers (PLB) were prepared by the Montal-Mueller technique in a FRAP system designed to simultaneously measure conductivity across, and lateral diffusion of, the bilayer. In the first stage of the project the FRAP system was used to characterise the lateral dynamics of bilayer lipids with regards to phospholipid composition (headgroup, chain unsaturation etc.), presence of cholesterol and the effect of divalent cations on negatively-charged bilayers. In the second stage of the project, lateral diffusion of two fluorescently-labelled voltage-dependent pore-forming peptides (alamethicin and S4s from Shaker K(+) channel) was determined at rest and in the conducting state. This study demonstrates the feasibility of such experiments with PLBs, amenable to physical constraints, and thus offers new opportunities for systematic studies of structure-function relationships in membrane-associating molecules. PMID- 12734603 TI - Bladder disease: A report based on the smooth muscle function in health and disease satellite meeting to the International Congress of Physiology. AB - One of the satellite meetings to the International Congress of Physiological Sciences concerned smooth muscle function in health and disease. In this report I have highlighted one of the areas of disease that were discussed: namely, bladder disease. In the treatment of unstable bladder, antimuscarinic agents are effective but cause unacceptable side effects. Thus, in order to prevent the side effects, new antimuscarinic agents are being prepared that show selectivity for the bladder and new methods for local delivery of antimuscarinics to the bladder are being developed. Potential new targets for the unstable bladder include P2X receptors and the nerve growth factor pathway. In detrusor-external sphincter dyssynergia, there is evidence to support a clinical trial of nitric oxide donors. The unpleasant and painful condition of interstitial cystitis has, to date, proven difficult to treat. Preliminary clinical data suggests that the leukotriene D(4) receptor antagonist montelukast may be useful in the treatment of IC. (c) 2001 Prous Science. All rights reserved. PMID- 12734602 TI - Recent advances in the field of platelet and bone marrow physiology and myeloproliferative and myelodysplastic diseases. PMID- 12734605 TI - Chemotherapy Foundation Symposium XIX: Innovative cancer therapy for tomorrow. PMID- 12734604 TI - Cancer therapy by gene therapy with angiostatin. AB - Angiostatin, an internal fragment of plasminogen, has been shown to inhibit angiogenesis. A new area of cancer research that has generated excitement is the use of angiostatin to treat cancer. Angiostatin protein therapy has not been pursued because current technology is inadequate to manufacture the needed biologically active proteins in sufficient quantities. It is sufficient for effective therapy with angiostatin to establish angiostatin production in the vicinity of tumors by gene transfer of angiostatin cDNA. There are various methods by which to transfer angiostatin cDNA. One way is to use a viral vector to incorporate the gene into cells. Another way is to use nonviral vectors. In this review, evidence accumulated from many laboratories suggests that angiostatin gene therapy may be an important new cancer therapy as an adjuvant therapy to prevent recurrence. (c) 2001 Prous Science. All rights reserved. PMID- 12734606 TI - CYP2C9 genotypes and diclofenac metabolism in Spanish healthy volunteers. AB - OBJECTIVE: This study analyzed the frequency of CYP2C9 variant alleles and evaluated the impact of CYP2C9 genotype on diclofenac metabolism in a Spanish population. METHODS: Diclofenac hydroxylation capacity was studied in a population of 102 healthy volunteers. After a single oral dose of 50 mg diclofenac the 0- to 8-h urinary concentrations of diclofenac and its main metabolites, 4'-hydroxy (OH), 3'-OH and 5-OH diclofenac were analyzed by high performance liquid chromatography. CYP2C9 genotyping for the variant alleles CYP2C9*2 and *3 was carried out with PCR-RFLP. RESULTS: The frequencies of CYP2C9*1, *2, and *3 alleles were 0.74 (95%CI: 0.68-0.80), 0.16 (95%CI: 0.11 0.21) and 0.10 (95%CI: 0.06-0.15), respectively, among the 102 Spaniards studied. The diclofenac/4'-OH diclofenac urinary ratio, but not the diclofenac/3'-OH diclofenac and diclofenac/5-OH diclofenac ratios, was related to CYP2C9 genotype. The diclofenac/4'-OH ratio was significantly higher among subjects with CYP2C9*1/*3 (0.83+/-0.4, n=14, 95% CI for the difference: 0.02-0.4) and CYP2C9*2/*3 (1.10+/-0.5, n=4, 95% CI for the difference: 0.16-0.8) genotypes compared to CYP2C9*1/*1 (0.62+/-0.3, n=59) and approximately threefold higher (1.8) in the only subject homozygous for CYP2C9*3 variant. CONCLUSIONS: The frequencies of CYP2C9*1, *2, and *3 alleles in the Spanish population reported here were similar to those found in the previously studied white European populations, and different of the previously reported in another Spanish population. CYP2C9*3 allele seems to influence the 4'-hydroxylation of diclofenac, although there is a large overlapping in the urinary metabolic ratio between the genotype groups studied PMID- 12734607 TI - A pilot double-blind randomized placebo-controlled study of molsidomine 16 mg once-a-day in patients suffering from stable angina pectoris: correlation between efficacy and over time plasma concentrations. AB - OBJECTIVES: A new once-a-day (o.a.d.) formulation of molsidomine (16 mg) was evaluated in patients with stable angina pectoris. The aims were to characterize its pharmacokinetics after a single dose, to demonstrate its clinical efficacy and safety versus placebo and to investigate correlations between pharmacokinetics and pharmacodynamics. METHODS: Forty-two patients were recruited in a double-blind, crossover, randomized placebo-controlled trial. The pharmacokinetics of molsidomine and SIN-1, its active metabolite, were determined at specific time points (3, 6, 10, 14, 18, 22 and 24 h) after the administration of a single dose of molsidomine 16 mg o.a.d. in all patients distributed into seven groups. Twenty-eight of these 42 patients showed a positive baseline cycloergometric exercise test response during the run-in placebo period and were used to compare the efficacy of molsidomine to placebo. Relationships between plasma concentration in molsidomine or SIN-1 and ischemic threshold were assessed in 16 of the 28 patients with a positive exercise test at baseline. Indeed, the censored variable ischemia-limited tolerance to exercise could not be evaluated in those patients who did not show exercise-induced ischemia anymore under molsidomine 16 mg o.a.d. Pharmacokinetic-pharmacodynamic relationships were evaluated using regression models and correlation coefficients. RESULTS: The highest average concentration in molsidomine and SIN-1 occurred after 6 h, then a plateau of 15-20 ng/ml molsidomine and 0.8-3.0 ng/ml SIN-1 was maintained for at least 8 h and the mean residual molsidomine concentration 24 h post-drug intake was around 8 ng/ml, still in the effective range of 5-10 ng/ml. A significant increase in total workload (+52 W min, P=0.009), total exercise time (+32 s, P=0.003) and time to angina (+25 s, P=0.016) was measured with molsidomine 16 mg o.a.d. relative to placebo. Using linear regression, significant correlation coefficients were determined between molsidomine plasma concentrations (but not SIN-1) and exercise test improvements (r=0.827, P<0.001 for the total workload; r=0.772, P<0.001 for the total exercise time; and r=0.566, P=0.028 for the time to 1 mm ST-segment depression). CONCLUSION: The pharmacokinetics of molsidomine 16 mg in patients with stable angina pectoris is compatible with a o.a.d. dosage regimen. This o.a.d. formulation is effective and well-tolerated, providing a 24 h therapeutic control of myocardial ischemia. A positive and significant linear relationship between molsidomine plasma concentration and the increase in exercise tolerance was observed. PMID- 12734608 TI - Pharmacokinetics and sedative effects in healthy subjects and subjects with impaired liver function after continuous infusion of clomethiazole. AB - OBJECTIVE: Clomethiazole is virtually completely eliminated by hepatic metabolism. This study was designed to assess the impact of liver impairment on its elimination and sedative effects. METHODS: Eight patients with mild liver impairment (Child-Pugh grade A), eight patients with moderate/severe liver impairment (Child-Pugh grade B/C) and eight healthy subjects of similar age were given 68 mg/kg clomethiazole edisilate according to a 24-h infusion scheme aimed at producing minimum sedation as it was intended for clinical use in patients with stroke. Concentrations of clomethiazole and its active alpha-carbon hydroxylated metabolite NLA-715 were followed in plasma and urine for 96 h and 24 h, respectively. Sedation was monitored using a scale from 1 to 6. RESULTS: The fraction excreted unchanged in urine was less than 0.2% for clomethiazole and less than 0.4% for NLA-715. Urine concentrations of clomethiazole were strongly correlated (r(2)=0.60) to plasma concentrations and approximately equal to unbound plasma concentrations. Plasma levels of NLA-715 increased steadily during the infusion, eventually reaching mean levels exceeding those of clomethiazole in all groups. Plasma clearance of clomethiazole in subjects with mildly impaired liver function was not statistically different from that of healthy controls (40 l/h vs 44 l/h). In subjects with moderate/severe liver impairment, there was a 50% reduction in clearance. Sedation was not observed except in two subjects in the Child-Pugh A group showing mild sedation. CONCLUSION: The reduced clomethiazole clearance in patients with moderate/severe liver impairment seems to call for a reduction of clomethiazole dosage. However, sedation was not observed in this group at the investigated dose level. PMID- 12734609 TI - Guidelines for iron supplementation in pregnancy: compliance among 431 parous Scandinavian women. AB - OBJECTIVE: The need for iron in pregnancy is rarely met by dietary food intake alone. Therefore, guidelines on iron supplementation have been developed to ensure optimal maternal and foetal iron provision. Today, the World Health Organization recommends iron supplementation during the second half of pregnancy. Our aim was to study compliance to these guidelines among 431 parous Scandinavian women. In addition, the association between maternal socio-demographic and lifestyle factors, maternal haemoglobin (Hb) values and non-compliance was assessed. METHODS: The women were interviewed about iron use at gestational weeks 25, 33 and 37 at the three Scandinavian study sites, Trondheim, Bergen and Uppsala. RESULTS: In all, 27% of the women used iron supplements continuously during the second half of pregnancy, hence complying with the guidelines. Further, 41% of the women reported periodical and 32% no use of iron supplementation during the second half of pregnancy. In the multivariable analyses, high Hb-concentration, living in Uppsala [OR: 3.1 (1.6-6.0)] or Bergen [OR: 4.7 (2.4-9.3)] as opposed to Trondheim, and smoking during pregnancy [OR: 2.0 (1.1-3.5)] were associated with non-compliance to guidelines. CONCLUSION: We conclude that maternal Hb values, demographic factors and smoking during pregnancy are among the factors associated with non-compliance to guidelines on iron supplementation during pregnancy. PMID- 12734612 TI - Grazing incidence small-angle X-ray scattering: an advanced scattering technique for the investigation of nanostructured polymer films. AB - Hamburg workshop on the "application of synchrotron radiation in chemistry"With grazing incidence small-angle X-ray scattering (GISAXS) the limitations of conventional small-angle X-ray scattering with respect to extremely small sample volumes in the thin-film geometry are overcome. GISAXS turned out to be a powerful advanced scattering technique for the investigation of nanostructured polymer films. Similar to atomic force microscopy (AFM), a large interval of length between molecular and mesoscopic scales is detectable with a surface sensitive scattering method. While with AFM only surface topographies are accessible, with GISAXS the buried structure is also probed. Because a larger surface area is probed, GISAXS also has a much larger statistical significance compared to AFM. Due to the high demand on collimation, GISAXS experiments are based on synchrotron radiation. Nanostructures parallel and perpendicular to the sample surface observable in thin poly(styrene- block-isoprene) diblock copolymer films are presented as an example of the possibilities of GISAXS. PMID- 12734611 TI - Hyponatremia associated with the initiation of reboxetine therapy. PMID- 12734610 TI - Inappropriate medication use among hospitalized older adults in Italy: results from the Italian Group of Pharmacoepidemiology in the Elderly. AB - OBJECTIVE: To determine the prevalence of inappropriate medication use among hospitalized older adults and to identify predictors of this use. METHODS: A total of 5734 patients (mean age 79 years) admitted to geriatric and internal medicine wards participating in the study in 1995 and 1997 were included in this analysis. Inappropriate medication use was defined on the basis of the criteria published by Beers in 1997. Only medications used during hospital stay were considered for the present study. RESULTS: During hospital stay, 837 (14.6%) patients received one or more medications classified as inappropriate based on Beers criteria. Ticlopidine ( n=346; 6.0% of the study sample) was the most frequently used medication among those in Beers' list, followed by digoxin ( n=174; 3.0%) and amytriptyline ( n=113; 2.0%). The multivariate analysis showed that age [75-84 years vs 65-74 years, odds ratio (OR) 0.85, 95% confidence interval (CI) 0.71-1.00; >or=85 years vs 65-74 years, OR 0.58, 95% CI 0.46-0.73], cognitive impairment (OR 0.77, 95% CI 0.64-0.94), Charlson co-morbidity index (>or=2 vs 0-1, OR 1.20, 95% CI 1.02-1.40) and overall number of medications used during hospital stay (5-8 medications vs <5 medications, OR 2.20, 95% CI 1.72 2.82; >or=9 medications vs <5 medications, OR 3.68, 95% CI 2.86-4.73) were significantly associated with use of inappropriate medications. CONCLUSIONS: Inappropriate medication use was common among hospitalized older adults. The most important determinant of risk of receiving an inappropriate medication was the number of drugs being taken. Older age and cognitive impairment were associated with a reduced likelihood of using an inappropriate medication. PMID- 12734614 TI - Biosorption of cadmium, copper, lead and zinc by inactive biomass of Pseudomonas Putida. AB - The accumulation of Cd(II), Cu(II), Pb(II) and Zn(II) at mg L(-1) concentration levels by inactive freeze-dried biomass of Pseudomonas Putida has been investigated. These metals could be efficiently removed from diluted aqueous solutions. A contact time of 10 min was sufficient to reach equilibrium. The pH has a strong effect on metal biosorption and the optimal pH values were 6.0, 5.0 6.0, 6.0-6.5 and 7.0-7.5 for Cd(II), Cu(II), Pb(II) and Zn(II) respectively. Under these conditions there was 80% removal for all metals studied. The process of biosorption can be described by a Langmuir-type adsorption model. This model accounts for 98% of the data variance. The K(A) and q(max) parameters for each metal are strongly correlated (at confidence levels greater than 98%) with the metal acidity, quantified by the constant of the corresponding M(OH)(+) complex, thus confirming previous assertions by other authors. PMID- 12734613 TI - Luminescence-based whole-cell-sensing systems for cadmium and lead using genetically engineered bacteria. AB - Whole-cell-based sensing systems that respond to cadmium and lead ions have been designed and developed using genetically engineered bacteria. These systems take advantage of the ability of certain bacteria to survive in environments polluted with cadmium and lead ions. The bacteria used in this investigation have been genetically engineered to produce reporter proteins in response to the toxic ions. This was achieved by modifying a strain of Escherichia colito harbor plasmids pYSC1 and pYS2/pYSG1. In these dual-plasmid-based sensing systems, the expression of the reporters beta-galactosidase and red-shifted green fluorescent protein (rs-GFP) was controlled by CadC, the regulatory protein of the cad operon. Regulation of the expression of the reporter proteins is related to the amount of cadmium and lead ions employed to induce the bacteria. The bacterial sensing systems were found to respond to cadmium, lead, and zinc ions, and had no significant response to nickel, copper, manganese, and cobalt. PMID- 12734615 TI - Free radical-scavenging activity of indolic compounds in aqueous and ethanolic media. AB - Indolic compounds are a broad family of substances present in microorganisms, plants and animals. They are mainly related with tryptophan metabolism, and present particularities that depend on their respective chemical structures. The most important members of the family are the plant hormone, indole-3-acetic acid, and the animal hormone, melatonin. An important characteristic of some indolic compounds is that they may be useful as chemical preventive agents against diseases such as cancer, oxidative stress, etc. For this reason, the possible antioxidant activities (free radical-scavenging activity) of several indoles were studied. The2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid /H(2)O(2)/HRP decoloration method was applied to determine both hydrophilic (in buffered media) and lipophilic (in organic media) antioxidant properties of the indolic compounds. Also, a study of the hydrophilic antioxidant activities of indoles at different pH values (between 4.5 and 8.5) was made. Finally, their possible role as diet plant antioxidants is discussed. PMID- 12734617 TI - Preliminary characterization of a light-rare-earth-element-binding peptide of a natural perennial fern Dicranopteris dichotoma. AB - A light-rare-earth-element (LREE)-binding peptide was isolated from LREE hyperaccumulator Dicranopteris dichotomaleaves and characterized in terms of molecular weight and ultraviolet absorption spectrum. The molecular weight of the LREE-binding peptide was determined to be 2208 Da by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOFMS). The characteristic ultraviolet absorption spectrum of the peptide was observed at 220 300 nm, suggesting that the peptide chain contained aromatic amino acids. Compared to the unique features of the phytochelatins with a low absorption at 280 nm, the LREE-binding peptide is unlikely to be a typical phytochelatin. The present study suggests that the LREE-binding peptide is probably a natural peptide in D. dichotoma, and it may play an important role in hyperaccumulation of LREEs. PMID- 12734616 TI - Flow injection analysis: Rayleigh light scattering technique for total protein determination. AB - A novel flow injection analysis (FIA) method with Rayleigh light scattering (RLS) detection was developed for the determination of total protein concentrations. This method is based on the weak intensity of RLS of bromothymol blue (BB) (3',3" dibromothymolsulfonephthalein) which can be enhanced by the addition of protein in weakly acidic solution. A common spectrofluorimeter was used as a detector. It was proved that the application of this method to quantify the total proteins in real samples by using bovine serum albumin was possible. The RLS signal was detected at lambda(ex)= lambda(em)=572 nm. The linear range was 7.0-70.0 microg mL(-1), the detection limit was 3.75 microg mL(-1), the reproducibility was 5.5% (n=7), and the sample throughput was 26 h(-1). PMID- 12734618 TI - One-step cleanup for PAH residue analysis in plant matrices using size-exclusion chromatography. AB - A new one-step cleanup procedure, based on size-exclusion chromatography (SEC), usable for the extracts from accelerated solvent extraction (ASE), Soxhlet extraction, or ultrasonic extraction (USE), is described. The method is suitable for the determination of polycyclic aromatic hydrocarbons (PAHs), especially from very complicated plant matrices (e.g. pine needles, deciduous leaves, mosses). The main improvement compared with previous conventional procedures is that analyte peaks barely overlap with matrix peaks in the chromatograms and that it is a very rapid and simple one-step procedure with clearly improved analytical performance. Essential advantages of this SEC procedure are the sharper GC-MS chromatograms for the PAH fraction at retention times between 9.2 and 12.0 min, distinctly separated substance peaks resulting in better analysis, shorter running times, and lower solvent consumption. PMID- 12734619 TI - Analysis of off-flavors in the aquatic environment by stir bar sorptive extraction-thermal desorption-capillary GC/MS/olfactometry. AB - The off-flavor compounds 2-methylisoborneol (MIB), geosmin, 2,4,6 trichloroanisole, 2,3,6-trichloroanisole, 2,3,4-trichloroanisole, and 2,4,6 tribromoanisole were analyzed in water samples by stir bar sorptive extraction (SBSE) followed by on-line thermal desorption (TD) capillary GC/MS. Quantification was performed using the MS in the single-ion-monitoring mode (SIM) with 2,4,6-trichloroanisole-D(5 )as internal standard. Quantification limits are 0.1-0.2 ng L(-1) for the haloanisoles, 0.5 ng L(-1) for geosmin, and 1 ng L(-1) for MIB. The relative standard deviations at the quantification limit ranged from 7 to 14.6%. SBSE recovery was evaluated by spiking real water samples and varied from 87 to 117%. More than twenty samples per day can be analyzed by SBSE-TD capillary GC-MS. The same technique in combination with olfactometry was used to elucidate unknown odorous compounds in water samples. PMID- 12734620 TI - Determination of inorganic selenium species by miniaturised isotachophoresis on a planar polymer chip. AB - The use of miniaturised isotachophoresis to allow the simultaneous determination of two inorganic selenium species has been investigated using a poly(methyl methacrylate) chip with a 44-mm-long, 200-microm-wide, 300-microm-deep separation channel. The miniaturised device included an integrated on-column, dual-electrode conductivity detector and was used in conjunction with a hydrodynamic fluid transport system. A simple electrolyte system has been developed which allowed the separation of selenium(IV) and selenium(VI) species to be made in under 210 s. The limits of detection were calculated to be 0.52 mg L(-1) for selenium(IV) and 0.65 mg L(-1 )for selenium(VI). The method allowed the separation of the selenium species from a range of common anions including fluoride, nitrate, nitrite, phosphate, sulfate and sulfite. PMID- 12734622 TI - High-performance liquid chromatography with diamond ATR-FTIR detection for the determination of carbohydrates, alcohols and organic acids in red wine. AB - A horizontal diamond attenuated total reflection (ATR) element has been incorporated in a flow-through cell with low dead volume and used for on-line mid IR detection in high-performance liquid chromatography. The chemical inertness of the ATR element permitted the use of a strongly acidic mobile phase in the isocratic separation. The hyphenation was used for the analysis of organic acids, sugars and alcohols in red wine. In the case of co-eluting analytes multivariate curve resolution-alternating least squares (MCR-ALS) was successfully employed for quantitative analysis. PMID- 12734621 TI - Effects of gamma-sterilization on butyltin homogeneity and content in sediments: a GC-ICP-MS study. AB - A GC-ICP-MS method based on extraction and alkylation of butyltins with sodium tetraethylborate was used to quantitatively assess the fate of these analytes in solutions and sediments following exposure to gamma-irradiation. The effects of a 2.5 Mrad sterilization dose on three butyltin species in both methanolic calibration solutions and in sediment matrices were investigated. Although significant losses of tributyltin (TBT, 90%), dibutyltin (DBT, 100%) and monobutyltin (MBT, 80%) were detected in standard solutions prepared in methanol following gamma-irradiation, no species inter-conversion occurred. Some degradation of TBT (38%) and DBT (32%) but no significant change in MBT content was found using a spiked sediment CRM HISS-1. Conversion DBT to MBT in spiked HISS-1 was deduced. Much smaller degradation of TBT (16% loss) and 10% loss of DBT by conversion to MBT (14% gain) was registered using a sediment blend of PACS 2 and HISS-1 (SOPH). Despite some initial losses of TBT and DBT due to irradiation, better than 2% RSD in both TBT and DBT concentrations measured in twelve different bottles of blended sediment SOPH were obtained, indicating the material may be considered homogeneous for these analytes. Results from a long term five-year stability study of PACS-2 show that all three butyltins are stable during storage at 4 degrees C followed with 2.5 Mrad minimum dose of gamma irradiation sterilization treatment. PMID- 12734623 TI - Determination of zearalenone and ochratoxin A in soil. AB - Mycotoxins are secondary metabolites, formed by the action of fungi on agricultural crops in the field or during storage. These metabolites are highly toxic to animals and humans and high levels have been measured in agricultural crops. In order to evaluate human risks due to ingestion of mycotoxin contaminated food different methods have been developed for analysis of mycotoxins in cereals and maize. In this project the focus was on mycotoxins in agricultural soil and the fate of these toxins in the soil-water-plant system. Two different mycotoxins were selected in the study: zearalenone (ZON) produced by species of Fusariumor Aspergillusand ochratoxin A (OTA) produced by species of Penicillium. We developed a method for analysis of these toxins in soil. Soil samples were extracted with methanol-water (9:1) and purified by solid-phase extraction (SPE, C8-columns). The final extract was analysed using high-pressure liquid chromatography (HPLC) with fluorescence detection. A Phenyl Hexyl column was used to separate the toxins. The detection limits obtained were 0.1 and 1.0 microg kg(-1) dry weight (dw) for OTA and ZON, respectively. The developed method has been used for analysis of different soils in connection with growth chamber experiments. The soil types used in the growth chamber experiments were a sandy soil, a sandy clay soil, and a soil with high content of organic matter. The recovery was determined as 85.8 and 93.4% and the repeatability to 5.1 and 12.8% for OTA and ZON, respectively. The reproducibility obtained was 8.5 and 15.0% for soil samples, representing concentration levels from 0.2-30 microg kg(-1) dw (OTA) and from 1.0-100 microg kg(-1) dw (ZON). PMID- 12734625 TI - Comparison of aqua regia and HNO3-H(2)O(2) procedures for extraction of Tl and some other elements from soils. AB - The relationship between aqua regia (ISO 11466) and HNO(3)-H(2)O(2) (ISO/CD 20279) extraction procedures for atomic emission spectrometric (ICP/OES and ICP/MS) determinations of Tl, P, Mn, Fe, Mg, Ca, Sr, Al, K, As, Bi, Zn, Pb, Co, Cd, Ni, V, Be, Cu and Cr was investigated. Soil samples (155) representing areas with different contents of the elements were selected for the comparison. Tl was the element of the highest interest and therefore the sampling sites were chosen to achieve as wide range of Tl contents as possible. Both extraction procedures are comparable in results (differences lower than 10% for the most of the elements) for all the tested elements. Statistically non-significant differences between the two extraction procedures were found for P, Zn, V and K (the slope was very close to 1 and the intercept included zero). Statistically significant values of intercepts were found for Fe, Al, Ca, Cd, Sr and Ni. Significantly higher results for aqua regia were found for Cu (12%), Pb (17%), Mn (11%) and lower results by aqua regia were found for Mg (4%), As (13%), Co (20%), Be (11%), Cr (4%) and Bi (6%). The results for Tl, the element of the highest interest, after HNO(3)-H(2)O(2) extraction procedure, were approximately 6% higher than the results after aqua regia extraction. Content of Tl in the soil samples was from 0.08 to 2.8 mg kg(-1). A highly significant linear relationship was found (R(2)=0.97). PMID- 12734624 TI - Evaluation of stability of arsenic species in rice. AB - Although most edible vegetables do not accumulate As at a high rate, rice, carrots and certain others are exceptions. In addition to nutritional or toxicological considerations, the relatively high level and variety of As species present in rice make it a very suitable matrix for a candidate reference material representative of terrestrial biological samples.An analytical procedure was developed for As speciation in rice based on the use of a 1:1 methanol-water mixture for species extraction, an anion Hamilton PRPX-100 column (at pH 6, and phosphate mobile phase 10 mM), and a cation Hamilton PRP-X200 column (at pH 2.8 in pyridine formiate 4 mM) for species separation and final determination by HPLC ICP-MS. The detection limits for dry flour rice expressed as As were 2 and 3 ng g(-1) for As(III) and AsB on the cation column and 3, 6 and 5 ng g(-1) for As(V), MMA and DMA, respectively, on the anion column. The methodology developed was applied to check the stability of As species in the water-methanol extract and also under different processing steps and storage time and temperature conditions. It was demonstrated that the As species in the water-methanol extracts stored at +4 degrees C remained stable for at least one month. Once the rice grains are ground, the MMA and As(V) species are not stable under any storage conditions probably due to microbiological activity. When ground rice is gamma-irradiated species remain stable although the AsB does not appear. PMID- 12734627 TI - Applications of capillary electrophoresis with laser-induced fluorescence for analysis of dGMP-BPDE adduct. AB - DNA adducts are thought to be crucial to the initiation of mutational and carcinogenic processes. Polycyclic aromatic hydrocarbons (PAHs) have been identified as one major source of carcinogenic risk since they can bind to DNA thus forming an adduct. Quantification of this adduct is important because it may correlate to the risk for cancer development. In this study, the adduct formed between 2'-deoxyguanosine 5'-monophosphate and benzo[ a]pyrene-7,8-dihydrodiol 9,10-epoxide (BPDE) was analyzed by capillary electrophoresis. Both capillary zone electrophoresis (CZE) and micellar electrokinetic capillary chromatography (MECC) modes with laser-induced fluorescence detection were used for the separation and analysis of DNA adducts. The exploration of capillary electrophoresis in several modes provided different separation mechanisms in which the stereochemical forms of the adduct could be separated. The best result obtained was using a coated fused-silica capillary in Tris-TAPS buffer, which provided high sensitivity with a detection limit of 2.5x10(-9) mol L(-1). MECC separation of the BPDE adduct, although less sensitive, provided an efficient enantioselective separation option. PMID- 12734626 TI - Separation of cocaine stereoisomers by capillary electrophoresis using sulfated cyclodextrins. AB - A capillary electrophoretic method for the separation of cocaine and its stereoisomers was developed. In this study, the effect of organic modifier was also investigated. The separation was achieved using 1% sulfated cyclodextrin, 10 mmol L(-1) phosphate buffer, 10% methanol at pH 3. The method provides good reproducibility and easy application. PMID- 12734629 TI - Speciation of arsenic in water, sediment, and plants of the Moira watershed, Canada, using HPLC coupled to high resolution ICP-MS. AB - High-performance liquid chromatography (HPLC) coupled with high-resolution sector field ICP-MS was applied to the speciation of arsenic in environmental samples collected from the Moira watershed, Ontario, Canada. Arsenic contamination in Moira River and Moira Lake from historic gold mine operations is of increasing environmental concern to the local community. In this study, the current arsenic contamination status in water, sediment, and plants was investigated. Elevated levels of arsenic in the surface water of up to 75 ng mL(-1) in Moira River and 50 ng mL(-1) in Moira Lake were detected, 98% of which was present as arsenate. High concentrations of arsenic (>300 ng mL(-1)), mainly present as arsenite, were detected in sediment porewaters. A sediment profile of As from the West basin of Moira Lake showed lower As concentrations compared with data from the 1990s. An optimized extraction procedure using a phosphoric acid-ascorbic acid mixture demonstrated that an unknown "As-complex" which may consist of As, sulfide and organic matter is potentially responsible for the release of arsenite from the sediment to the overlying water column. Arsenic concentrations in plant samples ranged from 2.6 to 117 mg kg(-1), dry weight. Accumulation of arsenic was observed in submerged plants collected from Moira River and Moira Lake. Only a small part of the arsenic (6.3-16.1%) in the plants was extractable with methanol water (9:1), and most of this arsenic (70-93%) was inorganic arsenic. A variety of organic arsenic compounds, including simple methylated compounds (methylarsonic acid and dimethylarsinic acid), trimethylarsine oxide, and tetramethylarsonium cation were detected at trace levels. No arsenobetaine and arsenocholine was found in any plant sample. An unknown compound, most probably an arsenosugar was detected in the two submerged plants, coontail ( Ceratophyllum demersum) and long-stemmed waterwort ( Elatine triandra). These submerged plants are constantly exposed to high arsenic concentrations in the surrounding water. Apparently, they are able to grow in this environment without invoking the same biochemical defence known from marine algae to detoxify inorganic arsenic. The detoxification mechanism of these plants remains unknown. PMID- 12734628 TI - A new approach to phosphoserine and phosphothreonine analysis in peptides and proteins: chemical modification, enrichment via solid-phase reversible binding, and analysis by mass spectrometry. AB - beta-Elimination of the phosphate group on phosphoserine and phosphothreonine residues and addition of an alkyldithiol is a useful tool for analysis of the phosphorylation states of proteins and peptides. We have explored the influence of several conditions on the efficiency of this PO(4)(3-) elimination reaction upon addition of propanedithiol. In addition to the described influence of different bases, the solvent composition was also found to have a major effect on the yield of the reaction. In particular, an increase in the percentage of DMSO enhances the conversion rate, whereas a higher amount of protic polar solvents, such as water or isopropanol, induces the opposite effect. We have also developed a protocol for enrichment of the modified peptides, which is based on solid-phase covalent capture/release with a dithiopyridino-resin. The procedure for beta elimination and isolation of phosphorylated peptides by solid-phase capture/release was developed with commercially available alpha-casein. Enriched peptide fragments were characterized by MALDI-TOF mass spectrometric analysis before and after alkylation with iodoacetamide, which allowed rapid confirmation of the purposely introduced thiol moiety. Sensitivity studies, carried out in order to determine the detection limit, demonstrated that samples could be detected even in the low picomolar range by mass spectrometry. The developed solid-phase enrichment procedure based on reversible covalent binding of the modified peptides is more effective and significantly simpler than methods based on the interaction between biotin and avidin, which require additional steps such as tagging the modified peptides and work-up of the samples prior to the affinity capture step. PMID- 12734631 TI - Solid-state NMR spectroscopy and its application in analytical chemistry. PMID- 12734630 TI - Development of a new microelectrolysis system for in-situ electrodeposition of ultra-traces of gold prior to measurement by ETAAS. AB - Electrodeposition is known to be suitable for separation and preconcentration of extremely low concentrations of analyte from bulk samples and is instrumentally is very simple. In this approach a new combined system was designed for in-situ electrodeposition of ultratrace levels of gold from micro samples on to a graphite furnace prior to determination by electrothermal atomic absorption spectrometry (ETAAS). Sediment samples were digested and traces of the gold content were extracted with boiling aqua regia. To prevent the highly corrosive effect of aqua regia media, the graphite tube surface was pre-coated by electrodeposition of ppm amounts of Pd prior to sample introduction. Separation of the analyte from the matrix was achieved by electrodeposition of the analyte in situ on the Pd/C surface of the furnace tube. Vanadium was also used as a modifier to stabilize the analyte from decomposition at charring temperatures. By using the proposed microelectrolysis preconcentration technique a considerable improvement in sensitivity and detection limit was achieved compared with conventional ETAAS. Characteristic masses for ED-ETAAS techniques in both nitric acid and aqua regia were reported to be m(o)=4.1 pg, with %RSD=2.9; the calculated LOD was 0.105 ppb, and typical calibration graphs for this element in nitric acid and aqua regia for both techniques were linear up to 70 microg L(-1) with about 99% recovery. Six reference sediments samples were tested by the proposed technique and by the Conv-ETAAS method. The results were in agreement with recommended values (reported by a reference laboratory using UV-visible spectrometry), demonstrating the efficiency of extraction and preconcentration of ultratrace levels of gold. PMID- 12734633 TI - New concepts in speciation analysis. PMID- 12734632 TI - Ionophore-based ion-selective potentiometric and optical sensors. AB - This paper describes ion-selective electrodes (ISEs) and optodes with a focus on the fundamental mechanisms, response characteristics, and recognition elements (ionophores) described to date for these sensors. The topics covered review the most frequently encountered ideas about ionophore-based sensors, including the theoretical and experimental parameters that influence response, typical ionophore structures, useful membrane compositions, and applications. Since these sensors are applicable in a number of fields, such as clinical, environmental, and process monitoring, as well as more novel approaches such as microfluidic based systems and micro/nanoprobes, this article provides descriptions that are oriented toward a more general audience. Furthermore, a large portion of this review describes the ionophore component itself in hopes of inspiring ideas in readers of how novel molecular architectures with new ionic targets or improved selectivity can be developed. PMID- 12734634 TI - Relative lack of cognitive effects of methylphenidate in elderly male volunteers. AB - RATIONALE: Methylphenidate, a dopaminergic and noradrenergic reuptake inhibitor, has been shown in young, healthy adult volunteers to produce pronounced effects on working memory and sustained attention. We were interested in assessing whether similar improvements could be conferred upon elderly volunteers in order to gain a more complete understanding of the effects of age on monoaminergic manipulations of working memory and attention, as well as to explore the potential for pharmacological intervention in attention and executive dysfunction disorders in this age group. OBJECTIVES: The main aim of the study was to characterise the dose-related effects of methylphenidate on a range of neuropsychological functions in elderly healthy volunteers. METHODS: Sixty healthy elderly adult male volunteers received either a single oral dose of placebo, 20 mg or 40 mg methylphenidate prior to performing a variety of tasks designed to assess memory, attention and executive function. A randomised double blind, between-subjects design was used. RESULTS: Methylphenidate had significant cardiovascular and subjective effects. However, unlike in younger volunteers, no significant effects of drug on working memory (spatial span and spatial working memory), response inhibition (stop-signal) or sustained attention (rapid visual information processing) were seen. Subtle effects on latency similar to those in younger volunteers were identified: both doses of methylphenidate resulted in a slowing in response time during set-shifting and decision-making. CONCLUSIONS: The results of this study demonstrate that, in elderly subjects, the cognitive effects of methylphenidate are grossly attenuated and distinct from the profile previously described in younger volunteers. It is suggested that methylphenidate may not be appropriate as a pharmacological intervention in elderly patient groups, such as those reporting age-related cognitive decline. PMID- 12734637 TI - Dermal and inhalation acute toxic class methods: test procedures and biometric evaluations for the Globally Harmonized Classification System. AB - The acute toxic class (ATC) methods were developed for determining LD(50)/LC(50) estimates of chemical substances with significantly fewer animals than needed when applying conventional LD(50)/LC(50) tests. The ATC methods are sequential stepwise procedures with fixed starting doses/concentrations and a maximum of six animals used per dose/concentration. The numbers of dead/moribund animals determine whether further testing is necessary or whether the test is terminated. In recent years we have developed classification procedures for the oral, dermal and inhalation routes of administration by using biometric methods. The biometric approach assumes a probit model for the mortality probability of a single animal and assigns the chemical to that toxicity class for which the best concordance is achieved between the statistically expected and the observed numbers of dead/moribund animals at the various steps of the test procedure. In previous publications we have demonstrated the validity of the biometric ATC methods on the basis of data obtained for the oral ATC method in two-animal ring studies with 15 participants from six countries. Although the test procedures and biometric evaluations for the dermal and inhalation ATC methods have already been published, there was a need for an adaptation of the classification schemes to the starting doses/concentrations of the Globally Harmonized Classification System (GHS) recently adopted by the Organization for Economic Co-operation and Development (OECD). Here we present the biometric evaluation of the dermal and inhalation ATC methods for the starting doses/concentrations of the GHS and of some other international classification systems still in use. We have developed new test procedures and decision rules for the dermal and inhalation ATC methods, which require significantly fewer animals to provide predictions of toxicity classes, that are equally good or even better than those achieved by using the conventional LD(50)/LC(50) methods. In order to cope with rather narrow dose/concentration classes of the GHS we have, as in our previous publications, combined the outcome of all results that can be obtained during testing for the allocation to one of the defined toxicity classes of the GHS. Our results strongly recommend the deletion of the dermal LD(50) and the inhalation LC(50) test as regulatory tests and the adoption of the dermal and inhalation ATC methods as internationally accepted alternatives. PMID- 12734635 TI - Methamphetamine attenuates disruptions in performance and mood during simulated night-shift work. AB - RATIONALE: Increased sleepiness while working and sleep disruptions are common complaints among shift workers. Consequently, shift workers may be more susceptible to diminished performance and work-related accidents. OBJECTIVE: To examine the effects of the central nervous system stimulant methamphetamine on psychomotor task performance, subjective effects, and food intake during shift work under laboratory conditions. METHODS: Seven participants completed this 23 day, within-participant design, residential laboratory study. They received a single oral methamphetamine dose (0, 5, 10 mg) 1 h after waking for three consecutive days under two shift conditions: (1) during the day shift, participants performed computerized psychomotor tasks from 0830 hours to 1730 hours and went to bed at 2400 hours and (2) during the night shift, participants performed tasks from 0030 hours to 0930 hours and went to bed at 1600 hours. Shifts alternated three times during the study; shift conditions were separated by an "off" day during which participants were not on a schedule and data were not collected. RESULTS: When participants received placebo, psychomotor task performance and subjective effects were disrupted during the night shift, relative to the day shift. Changing shift conditions did not alter food intake significantly. Methamphetamine reversed performance and subjective-effects disruptions, and decreased food intake during the night shift. CONCLUSION: These data indicate that shift changes produce performance impairments and mood alterations, and that a single low to moderate dose of methamphetamine attenuates many shift change-related disruptions in performance and mood. PMID- 12734636 TI - 9-Ethyladenine derivatives as adenosine receptor antagonists: 2- and 8 substitution results in distinct selectivities. AB - 9-Ethyladenine was used as the basis for a series of non-xanthine adenosine receptor antagonists at human adenosine receptors. The adenine-based compounds were substituted in 2- or 8-position with a variety of side chains including some aryl or arylalkynyl groups previously tested as 2-substituents in adenosine and 5'-N-ethylcarboxamidoadenosine (NECA) for their effect on agonist affinity. The affinity of the novel compounds was tested in radioligand binding assays (A1, A2A and A3) and inhibition of NECA-stimulated adenylyl cyclase activity (A2B) in membranes prepared from CHO cells stably transfected with the respective human receptor subtype. High affinity antagonists were identified for A1 (9-ethyl-8 phenyl-9H-adenine, compound 2; 6-(1-butylamino)-9-ethyl-8-phenyl-9H-purine, compound 3), A2A (8-ethoxy-9-ethyladenine; compound 8) and A3 (9-ethyl-8 phenylethynyl-9H-adenine, compound 5) with selectivities versus other receptor subtypes in the range of 10 to 600. These results demonstrate that adenine is a useful template for further development of high-affinity antagonists with distinct receptor selectivity profiles. PMID- 12734638 TI - In vitro dermal penetration study of carbofuran, carbosulfan, and furathiocarb. AB - In this study, the dermal penetration rate of carbofuran, carbosulfan, and furathiocarb has been measured with rat abdominal skin using the static diffusion cell. The technical grades of three compounds were applied at different doses on skin surface mounted in static diffusion cell and incubated at 32 degrees C for 48 h with shaking. The same procedures were carried out with furathiocarb EC (emulsifiable concentrate) and WP (wettable powder). At regular intervals, the receptor fluid in cell was sampled and analyzed by HPLC. Only carbofuran was found in carbosulfan- or furathiocarb-treated samples, suggesting they converted into carbofuran while passing through the skin layer. The quantity of insecticide penetrating skin increased with time and applied dose. The skin penetration rate increased with the water solubility of insecticides. The dermal penetration rates of carbofuran, furathiocarb, and carbosulfan were determined as 1.05 micro g/cm(2) per h ( r(2)=0.991), 0.46 micro g/cm(2) per h ( r(2)=0.984) and 0.14 micro g/cm(2) per h ( r(2)=0.967), respectively. There was no significant difference in rate of skin penetration between furathiocarb EC (1.42 micro g/cm(2) per h, r(2)=0.988) and WP (1.35 micro g/cm(2) per h, r(2)=0.982), while furathiocarb technical grade showed a lower skin penetration rate. In vitro models may be used to predict percutaneous absorption and are useful in selecting safer formulations for field application of pesticide. PMID- 12734639 TI - Contribution of CYP2E1 to N-methyl-2-pyrrolidone metabolism. AB - The involvement of cytochrome P450 2E1 (CYP2E1) in the metabolism of N-methyl-2 pyrrolidone (NMP) was studied with three experimental approaches: in the rat, in vitro in human microsomes, and in human volunteers. NMP was administered dermally (40 mg/kg) to OFA rats to examine the influence of CYP2E1 inhibition (5 mg/kg diethyldithiocarbamate, DETC, 30 min before) and CYP2E1 induction (after 4 days of fasting). The main NMP metabolite 5-hydroxy- N-methylpyrrolidone (5HNMP) in the urine fractions collected during the following 48 h was analysed by gas chromatography-mass spectrometry. CYP2E1 inhibition led to a statistically significant retardation of 5HNMP excretion in urinary fractions collected during the first 12 h. In the group of fasted rats, a two-fold increase of CYP2E1 activity was observed in comparison with the control group. During the first 6 h after dermal administration of NMP to fasted rats, about 33% of the dose was excreted in urine versus 22% in controls. In vitro, NMP (15 mM) was incubated (up to120 min) with human liver microsomes and the formation of 5HNMP followed Michaelis-Menten kinetics with V(max) of 1.1 nmol/min per mg protein and K(m) of 2.4 mM. The formation of 5HNMP was inhibited by 35% in the presence of a monoclonal antibody against CYP2E1, but not by CYP1A2 antibody. In a dermal application experiment, 12 humans volunteers were exposed by means of a dermal patch to 300 mg NMP; five urine fractions were collected during the 48 h following the onset of application in order to measure the major metabolites 5HNMP and 2-hydroxymethylsuccinimide (2HMSI). Before NMP application, a blood sample was collected for the quantification of CYP2E1 mRNA in peripheral blood lymphocytes (PBLs). The mean dermal absorption of NMP was 67.9%. The highest amount of 5HNMP was excreted in urine in the fraction collected between 6-12 h (12.6% of dose), while 2HMSI peaked in fractions 12-24 h and 36-48 h (3.3 and 3.2% of dose, respectively). A significant relationship was found between CYP2E1 mRNA content in PBLs and the amount of both the metabolites excreted in urine within 24 h ( r(2)=0.54, P<0.01). It is concluded that CYP2E1 is involved in the first steps of NMP metabolism in the rat and, to a lesser extent, in humans. Since large variations in CYP2E1 activity exist in the human population (at least 5-fold range), it seems justified to take into account the activity of this enzyme in an individual for an accurate interpretation of biological monitoring of exposure to NMP when relying on 5HNMP and/or 2HMSI determination in urine. PMID- 12734640 TI - Suppression of erythropoietin gene expression by cadmium depends on inhibition of HIF-1, not stimulation of GATA-2. AB - Long-term exposure of rats to cadmium (Cd) resulted in a marked suppression of erythropoietin (Epo) mRNA expression in the kidneys and the development of severe anemia. A recent report revealed that Cd inhibited hypoxia-inducible factor 1 (HIF-1) binding activity and Epo mRNA expression and protein production. However, Epo gene expression is also regulated by transcription factor GATA-2, which binds to the GATA binding site of the Epo promoter. To elucidate the mechanism of suppression of Epo by Cd, the effect of Cd on GATA-2 function was studied. Epo promoter/enhancer luciferase constructs, one with the wild-type promoter and another with a promoter with a mutant GATA site, were transfected into Hep3B cells. No significant difference in Epo promoter activity in these two types of cells was observed in the presence of Cd. The binding activity of GATA-2 was not affected by Cd. This study showed that Cd inhibited HIF-1 binding activity and Epo promoter activity, and then suppressed Epo protein production. Inhibition of Epo gene expression by Cd depends on suppression of HIF-1 binding activity, not on alteration of GATA function. PMID- 12734641 TI - Development of a high-performance reporter plasmid for detection of chemicals with androgenic activity. AB - A number of chemicals are present in the environment, and some synthetic chemicals may disrupt endocrine function of wild animals and humans. An effective procedure to screen chemicals for endocrine modulating activity has been needed to ensure the safety of chemicals, and the reporter gene assay technique may provide a powerful tool for screening endocrine-disrupting chemicals. We have developed a high-performance reporter plasmid that can trigger high androgen dependent induction with high selectivity by using mouse mammary tumor virus (MMTV) androgen-responsive elements and a partial fragment of the rat alpha(2u) globulin promoter region. This new type plasmid can induce higher transcriptional activation than a commercial PGV-P-based construct bearing the SV40 promoter fragment, and the basal induction level of this plasmid is much lower than that of the PGV-P-based construct. Moreover, only androgen derivatives could selectively induce a high response in the reporter gene assay with the new reporter plasmid. This new type of reporter plasmid, ARE-AUG- Luc+, should be of value in endocrine research and in screening to identify endocrine-modulating chemicals. PMID- 12734642 TI - 2,3,7,8-tetrachlorodibenzo- p-dioxin (TCDD) induces oxidative stress in the epididymis and epididymal sperm of adult rats. AB - 2,3,7,8-Tetrachlorodibenzo- p-dioxin (TCDD) is one of the most potent environmental contaminants, which has been shown to induce oxidative stress in testis and epididymal sperm of rats. However, the nature and mechanism of action of TCDD on the epididymis is not clear. The aim of the present study was to investigate whether induction of oxidative stress in epididymal sperm was direct effect of TCDD on epididymis. In the present studies, TCDD (0.1, 1.0 and 10 micro g/kg body weight per day) was administered orally to rats for 4 days. Twenty-four hours after the last treatment the animals were killed using anesthetic ether. Both epididymides were dissected out and epididymal sperm were collected by cutting the epididymides into small pieces in Ham's F-12 medium at 35 degrees C. The epididymal sperm and caput, corpus and cauda epididymides were homogenized and used for biochemical studies. Epididymal sperm counts did not decrease in the rats treated with TCDD. Administration of TCDD increased the production of reactive oxygen species such as hydrogen peroxide while the activities of antioxidant enzymes superoxide dismutase, catalase, glutathione reductase and glutathione peroxidase were found to be decreased in the epididymal sperm as well as in cauda epididymides. Lipid peroxidation also increased in the epididymal sperm and in the various regions of the epididymides after exposure to TCDD. The results indicated that TCDD induces oxidative stress in the epididymis and epididymal sperm by decreasing the antioxidant enzymes through induction of reactive oxygen species. Thus, the adverse effects of TCDD on the epididymal sperm were due to direct effect of TCDD on epididymis. PMID- 12734643 TI - Protective effects of glutathione on 5-fluorouracil-induced myelosuppression in mice. AB - The protective effects of glutathione (GSH) administration on myelosuppression induced by 5-fluorouracil (5-FU) were investigated in female BALB/c mice. Animals were allocated to four groups (16 mice/group). GSH was given orally at a dose of 800 mg/kg to groups 3 and 4 for 21 consecutive days (day 0 to day 20). 5-FU was repeatedly administered at a dose of 40 mg/kg to groups 2 and 3 for 1 week (day 7 to day 13) by gavage. Group 3 served as a combined treatment group and group 1 as a non-treated control group. The total observation period was 3 weeks. Body weight was measured once a week. A decrease in body weight due to 5-FU treatment was observed in groups 2 and 3 on day 14. Although the body weight in group 2 had not increased by 1-week after cessation of 5-FU treatment, the value in group 3 markedly recovered. Hematology, total nucleated myelocyte count and histopathology of bone marrow were carried out on day 14 and day 21. In groups 2 and 3, these examinations showed thrombocytopenia, leukopenia, reticulocytopenia and myelosuppression on day 14. However, platelets and bone marrow were less affected in group 3 than in group 2. On day 21, the thrombocytopenia in groups 2 and 3 was resolved. The myelosuppression, leukopenia and reticulocytopenia resolved in group 3, but not in group 2. Although simple microcytic anemia occurred delayed on day 21, it was less severe in group 3 than in group 2. Therefore, GSH may have preventive effects against 5-FU-induced hematopoietic toxicity, and accelerate recovery after cessation of 5-FU treatment. PMID- 12734644 TI - High-dose clastogenic activity of aniline in the rat bone marrow and its relationship to the carcinogenicity in the spleen of rats. AB - To clarify the question of clastogenicity of aniline in rats two studies were performed: a bone marrow micronucleus test and a bone marrow metaphase test. In the micronucleus test aniline (as aniline hydrochloride) was administered to groups of seven male PVG rats at single oral doses of 0, 300, 400, or 500 mg/kg body weight. Bone marrow was obtained 24 and 48 h after oral treatment. Smears of bone marrow were stained with acridine orange and erythrocytes were examined for the presence of micronuclei. Animals receiving cyclophosphamide (1x7.5 mg/kg) served as positive controls. Clinical signs observed in animals dosed at 300 mg/kg and above included cyanosis, light brown coloured urine and cold to touch. Small, but statistically significant and dose-related increases in the incidence of micronulei over the vehicle control values were observed at the 24-h sampling time only. Cyclophosphamide induced a significant and comparably much higher increase in micronuclei than aniline. In the bone marrow metaphase test aniline (as aniline hydrochloride) was administered to groups of seven male PVG rats at single oral dose levels of 0, 300, 400, or 500 mg/kg body weight. Bone marrow was sampled 18 and 30 h after dosing. A group treated with cyclophosphamide (1x40 mg/kg) served as positive control. A small increase in the percentage of aberrant cells above solvent control values was recorded in one rat at 400 mg/kg and four rats at 500 mg/kg at the 18-h sampling time only. The positive control cyclophosphamide induced a much higher rate of aberrant cells in all animals. Several lines of evidences are presented against a causal relationship between the clastogenic activity in male PVG rats at 400 and 500 mg/kg and the carcinogenicity in the spleen of Fischer 344 rats starting at 30 mg/kg in males. Among these are the dose-response relationship of the tumour incidence, the close correlation between degree of spleen damage and tumour induction, the lack of carcinogenic effects in mice even at higher dose levels, or in rats at dose levels inducing only slight haematotoxicity and spleen toxicity, and the available data on the mode of action of other chemicals inducing spleen tumours. PMID- 12734645 TI - Effects of the mycotoxin ochratoxin A in a bacterial and a mammalian in vitro mutagenicity test system. AB - Ochratoxin A (OTA), a mycotoxin produced by several Aspergillus and Penicillium species, is a worldwide contaminant of food and feedstuffs. It is nephrotoxic, immunosuppressive and carcinogenic in several animal species. The mechanism by which OTA acts is not fully understood up to now. Here, OTA was evaluated for mutagenicity in the Salmonella typhimurium assay (Ames assay) and in the HPRT assay with V79 hamster fibroblasts. In the bacterial assay using the strains TA 98, TA 100, TA 1535, TA 1538, TA 102 and TA 104, OTA was not mutagenic at a concentration range from 0.01 to 500 micro M in the presence and absence of an external metabolising enzyme system (rat liver S9 enzyme mix). In V79 fibroblasts, cytotoxicity of OTA was estimated with the neutral red uptake assay. An IC(50) of 11.6 micro M was found in the absence and an IC(50) of 6.4 micro M in the presence of S9 mix. In the subsequent HPRT (hypoxanthine-guanine phosphoribosyl-transferase) assay with V79 cells the negative result of the bacterial assay was confirmed using OTA in concentrations from 0.1 to 100 micro M. In order to obtain converted OTA metabolites from viable, metabolically competent cells, a preincubation of primary cultured rat hepatocytes with 0.016 to 0.8 micro M OTA was performed. The resulting culture medium, which contained OTA metabolites, was tested in both mutagenicity assays. Again, no mutagenic effect was detected either in the bacterial or in the mammalian test assay. In accordance with several literature data, the present results imply that OTA does not act as direct mutagen. Additionally, the OTA metabolites derived from cultured rat hepatocytes or rat liver S9 mix, also, do not have a mutagenic potency in the test systems used. PMID- 12734646 TI - A single-centre experience of post-renal transplant lymphoproliferative disorder. AB - Post-transplant lymphoproliferative disorder (PTLD) complicates 1 to 10% of all transplantations. Previous clinicopathological studies of PTLD have been limited by small numbers, short follow-up times, outdated data, heterogeneity of pooled solid-organ transplant results, and selective inclusion of early-onset disease. We therefore undertake here a retrospective analysis and identify all cases of PTLD that complicated renal transplantation at the Princess Alexandra Hospital between 30 June 1969 and 31 May 2001. Tumour samples were subsequently retrieved for pathological review and for Epstein-Barr virus-encoded RNA in situ hybridisation (EBER-ISH). Of 2,030 renal transplantation patients, 29 (1.4%) developed PTLD after a median period of 0.5 years (range 0.1 to 23.3 years). PTLD patients were more likely to have received cyclosporine (76% versus 62%, P<0.05), tacrolimus (10% versus 2%, P<0.05) and OKT3 (28% versus 10%, P<0.01). As the burden of immunosuppression increased from dual, to triple, to OKT3 therapy, the risks of early onset, extensive-stage, polymorphic, Epstein-Barr virus (EBV) associated and fatal PTLD progressively increased. The majority of patients presented with an extra-nodal mass (45%), were afebrile (76%), and had stage-IV disease (60%). EBER-ISH was positive in 58%. Actuarial 5-year disease-free survival was 53.7%. The independent predictors of mortality on multivariate Cox regression were polymorphic histology (HR 7.4, 95% CI 1.5-37) and an international prognostic index (IPI) >1 (HR 2.7, 95% CI 1.1-6.8). Compared with other treatments, chemotherapy was associated with higher survival rates (100% versus 18% at 3 years, P=0.0001). In conclusion, PTLD is more likely, occurs earlier, and is more often fatal, in the setting of intensive immunosuppression. Nevertheless, excellent long-term outcomes are achievable with early recognition and institution of appropriate treatment. PMID- 12734648 TI - Acute respiratory failure after a tiger mosquito bite: a case report. PMID- 12734647 TI - Endothelin receptor blockade in canine oleic acid-induced lung injury. AB - OBJECTIVE: To investigate the effects of endogenous endothelins on pulmonary haemodynamics and gas exchange in oleic acid lung injury. DESIGN: Prospective experimental study in dogs. SETTING: Animal research laboratory in a university teaching hospital. SUBJECTS. Seventeen anaesthetised and ventilated mongrel dogs. INTERVENTIONS: Nine pretreated dogs received an infusion of the endothelin A and B receptor antagonist bosentan (10 mg/kg) started before oleic acid. Eight treated dogs received bosentan started 90 min after oleic acid. Cardiac index (CI) was manipulated by inflating an inferior vena caval balloon or by opening a femoral arterio-venous bypass. MEASUREMENTS AND RESULTS: Pulmonary vascular resistance was defined by measuring the gradient between mean pulmonary artery pressure (MPAP) and occluded PAP (PAOP) at five levels of CI. Intrapulmonary shunt was measured using the inert gas SF(6). Pretreatment with bosentan prevented the oleic acid-induced shift of (MPAP-PAOP)/CI plots to higher pressures, but did not affect the increase in intrapulmonary shunt. Treatment of established oleic acid lung injury with bosentan had no effect. CONCLUSIONS: Pretreatment, but not treatment, with bosentan, in the dose used, blunted the oleic acid-induced increase in pulmonary vascular resistance, suggesting that endothelins contribute to the increase in pulmonary vascular tone in the early stages of oleic acid lung injury. PMID- 12734650 TI - Clostridium difficile colitis causing toxic megacolon, severe sepsis and multiple organ dysfunction syndrome. PMID- 12734649 TI - Ventilator-induced heat shock protein 70 and cytokine mRNA expression in a model of lipopolysaccharide-induced lung inflammation. AB - OBJECTIVE: To investigate the effect of mechanical ventilation with no PEEP (ZEEP) and 4 cmH(2)O PEEP on heat shock protein 70 (HSP70) and pulmonary inflammatory cytokine expression in a model of lipopolysaccharide (LPS) induced lung inflammation. DESIGN AND SETTING: Prospective, randomized, experimental animal study. SUBJECTS AND INTERVENTIONS: We challenged 42 male Sprague-Dawley rats intratracheally with LPS. After 24 h the rats were randomly assigned to one of the ventilation strategies. Rats received either 4 h of mechanical ventilation with ZEEP or mechanical ventilation with 4 cmH(2)O PEEP. A nonventilated control group received LPS only. Lung pathology after LPS challenge was evaluated by histology to assess baseline lung injury. HSP70 and cytokine mRNA levels were measured in total lung homogenates. RESULTS: PaO(2) levels and lung histology revealed no deterioration after PEEP ventilation and severe deterioration after ZEEP ventilation. There was a significant higher expression of HSP70 and IL-1beta mRNA in the lungs of the ZEEP group than in the PEEP group and nonventilated controls. In the ZEEP group high HSP70 levels were correlated inversely with low IL-1beta mRNA and low IL-6 mRNA. CONCLUSIONS: We propose that HSP70 expression protects the lung against ventilator-induced lung injury by decreasing cytokine transcription in the lung. PMID- 12734651 TI - Comparison of the Glasgow Coma Scale and the Reaction Level Scale for assessment of cerebral responsiveness in the critically ill. AB - OBJECTIVE: The Glasgow Coma Scale (GCS) is a well-known source of error in outcome prediction models. We compared assessment of cerebral responsiveness with the GCS and the Reaction Level Scale (RLS) in two otherwise similar outcome prediction models. DESIGN AND SETTING: Prospective, observational study in a general intensive care unit. PATIENTS AND PARTICIPANTS: All admissions of patients with or at risk of developing impaired brain function between 1997 and 1998 ( n=534). MEASUREMENTS AND RESULTS: Admissions were scored by RLS and APACHE II (includes scoring with the GCS). The RLS scores were transformed to APACHE II central nervous system scores according to a predetermined protocol. APACHE II estimated probability of death was calculated conventionally with the GCS and the RLS. Vital status 90 days after admission was secured from a national database. Bias and precision was 0.5% and 16.6%, respectively. The area under receiver operating characteristic curves was slightly but significantly greater with the RLS-based APACHE II model than with the GCS-based model (0.92 vs. 0.90). Discrimination was improved primarily in admissions with low and intermediate probability of death. CONCLUSIONS: Scoring of cerebral responsiveness with the RLS instead of the GCS was associated with minimal bias of the APACHE II probability of death estimate. Assessment of consciousness in critically ill with the RLS deserves further evaluation PMID- 12734652 TI - A readers' guide to the interpretation of diagnostic test properties: clinical example of sepsis. AB - BACKGROUND: One of the most challenging practical and daily problems in intensive care medicine is the interpretation of the results from diagnostic tests. In neonatology and pediatric intensive care the early diagnosis of potentially life threatening infections is a particularly important issue. FOCUS: A plethora of tests have been suggested to improve diagnostic decision making in the clinical setting of infection which is a clinical example used in this article. Several criteria that are critical to evidence-based appraisal of published data are often not adhered to during the study or in reporting. To enhance the critical appraisal on articles on diagnostic tests we discuss various measures of test accuracy: sensitivity, specificity, receiver operating characteristic curves, positive and negative predictive values, likelihood ratios, pretest probability, posttest probability, and diagnostic odds ratio. CONCLUSIONS: We suggest the following minimal requirements for reporting on the diagnostic accuracy of tests: a plot of the raw data, multilevel likelihood ratios, the area under the receiver operating characteristic curve, and the cutoff yielding the highest discriminative ability. For critical appraisal it is mandatory to report confidence intervals for each of these measures. Moreover, to allow comparison to the readers' patient population authors should provide data on study population characteristics, in particular on the spectrum of diseases and illness severity. PMID- 12734654 TI - Microsatellite analysis of ancient alpine grape cultivars: pedigree reconstruction of Vitis vinifera L. 'Cornalin du Valais'. AB - Ancient and closely related grape cultivars from the Alps were analyzed with 50 microsatellite markers: 'Cornalin', 'Humagne Rouge' and 'Goron' from Valais (Switzerland); 'Cornalin', 'Petit Rouge' and 'Mayolet' from the Aosta Valley (Italy). Our results confirmed previous studies showing that the 'Cornalin' cultivars from Switzerland and Italy are distinct, and that 'Humagne Rouge' is identical to 'Cornalin' from the Aosta Valley. We propose the nomenclature 'Cornalin du Valais' and 'Cornalin d'Aoste' in order to prevent further confusion. At each locus, 'Goron', 'Petit Rouge', 'Mayolet' and 'Cornalin d'Aoste' all share at least one allele with 'Cornalin du Valais', strongly suggesting parent/offspring relationships. Alleles at 49 out of 50 microsatellite loci are consistent with 'Cornalin du Valais' being the progeny of 'Petit Rouge' and 'Mayolet'. The exception is a 10-base pair discrepancy at one locus, most likely the result of somatic mutation in one of the parents, since this parentage is supported by high likelihood ratios and historical data. We hypothesize that 'Cornalin du Valais' originated in the Aosta Valley through a natural cross and was then introduced into Valais centuries ago, probably via the Great St. Bernard Pass. Furthermore, 'Cornalin du Valais' is likely to be one of the parents of both 'Goron' and 'Cornalin d'Aoste', the respective second parents remaining unknown. This pedigree provides a convincing explanation for the allele-sharing patterns and is strongly supported by historical data. The present work is the first grapevine parentage study to deal with a multiple repeat unit discrepancy at a microsatellite locus. We suggest that the use of increasingly large numbers of loci in making parentage determinations leads to a corresponding increase in the probability of encountering a locus with intra-cultivar variability during the analysis. We therefore assume that a sole multiple repeat unit discrepancy is not sufficient to discard a parentage hypothesis. PMID- 12734653 TI - Quality of life on admission to the intensive care: can we query the relatives? AB - OBJECTIVE: To study whether the Short Form-36 questionnaire can be used to assess the patient's quality of life on admission to the ICU by use of proxies in both scheduled and emergency admissions. DESIGN AND SETTING: Prospective study involving direct interviews of patients and relatives before or during ICU stay in a 10-bed mixed intensive care unit in a 654-bed university affiliated hospital. PATIENTS AND PARTICIPANTS: Patients before major elective surgery ( n=55) or following emergency admissions ( n=57). MEASUREMENTS AND RESULTS: Patients and proxies completed a health questionnaire in the first 72 h following emergency admission or the day before a scheduled admission to the ICU. Internal consistency was evaluated by measurement of Cronbach's alpha. All dimensions of the SF-36 had adequate internal consistency. On all eight dimensions a significant correlation was found between the patient and their proxy. In general, proxies underestimated the patient's quality of life although differences were small (less than 5%). On most items a good to very good agreement was found (alpha>0.6). Quality of life assessment was not affected by the admission status of the patient (acute or elective admission and surgical or medical diagnosis). CONCLUSIONS: The SF-36 questionnaire completed by a proxy can reliable assesses the quality of life of the critically ill patient on admission to the ICU. Proxies underestimated the patient's quality of life, although the differences were small. PMID- 12734655 TI - Mapping genes affecting flowering time and frost resistance on chromosome 5B of wheat. AB - Two populations of single chromosome recombinant lines were used to map genes controlling flowering time on chromosome 5B of wheat, and one of the populations was also used to map a new frost resistance gene. Genetic maps were developed, mainly using microsatellite markers, and QTL analysis was applied to phenotypic data on the performance of each population collected from growth-room tests of flowering time and frost tolerance. Using a recombinant substitution-line mapping population derived from a cross between the substitution-line 'Chinese Spring' ('Cheyenne' 5B) and 'Chinese Spring' (CS), the gene Vrn-B1, affecting vernalization response, an earliness per se locus, Eps-5BL1, and a gene, Fr-B1, affecting frost resistance, were mapped. Using a 'Hobbit Sib' ('Chinese Spring' 5BL) x 'Hobbit Sib' recombinant substitution line mapping population, an earliness per se locus, Eps-5BL2 was mapped. The Vrn-B1 locus was mapped on the distal portion of the long arm of chromosome 5B, to a region syntenous with the segments of chromosomes 5A and 5D containing Vrn-A1 and Vrn-D1 loci, respectively. The two Eps-5BL loci were mapped close to the centromere with a 16 cM distance from each other, one in agreement with the position of a homoeologous locus previously mapped on chromosome 5H of barley, and suggested by the response of 'Chinese Spring' deletion lines. The Fr-B1 gene was mapped on the long arm of chromosome 5B, 40 cM from the centromeric marker. Previous comparative mapping data with rice chromosome 9 would suggest that this gene could be orthologous to the other Fr genes mapped previously by us on chromosomes 5A or 5D of wheat, although in a more proximal position. This study completes the mapping of these homoeoallelic series of vernalization requirement genes and frost resistance genes on the chromosomes of the homoeologous group 5 in wheat. PMID- 12734656 TI - Mutation detection in rice waxy mutants by PCR-RF-SSCP. AB - PCR-RF-SSCP (PRS), which combines cleaved amplified polymorphic sequence (CAPS) and single-strand conformation polymorphism (SSCP), is expected to be a useful technique for DNA polymorphism analysis. We evaluated the ability of PRS to detect single nucleotide polymorphism (SNP) using the Waxy gene, Wx, of rice, and subsequently were able to identify point mutations in wx mutant lines. The approximately 6-kb Wx gene was divided into five regions for PCR amplification. Two regions, in which most of the point mutations of the wx mutants have been identified, were amplified by PCR and cloned into a vector, and those clones containing SNPs produced as a result of the inherent inaccuracy of PCR were used for the evaluation of PRS. The efficiency of PRS in the detection of SNPs of these clones was over 70%. PRS analysis of the wx genes in 18 waxy mutants was carried out in the five regions using two different restriction endonucleases and two gel conditions, with and without glycerol. Of the 18 lines tested, 17 showed band patterns different from that of the wild type. Most of the mutations identified in this study were nucleotide changes in exons, which result in amino acid changes. One mutation generated an in-frame stop codon, and another was a frame shift mutation by one-base deletion. Two mutations found at a splice site were considered to inhibit normal splicing of mRNA. These results show that PRS is a useful technique for detecting point mutations in large plant genes. PMID- 12734658 TI - Analysis of TP53 and PTEN in gliomatosis cerebri. AB - Gliomatosis cerebri (GC) is a rare glial neoplasm with extensive diffuse brain infiltration but relative preservation of the underlying architecture. Previous molecular studies, mostly analyzing biopsy samples, have suggested an astrocytic origin of GC, but a larger collective of autopsy tissue has not been investigated so far. Furthermore, whether the widespread neoplastic infiltration is based on a monoclonal process is still a matter of debate. In the present study, we screened paraffin-embedded brain tissue from different areas of 18 cases (8 autopsy cases and 10 biopsies) for alterations in the TP53 and PTEN genes. Nuclear accumulation of p53 protein was detected in 9 cases (50%). Somatic TP53 mutations occurred in two autopsy cases (11% of all cases). In the first case, a C-->T transition in codon 273 (Arg-->Cys) was detected in all tumor samples. In the second case, in tumor samples from one hemisphere, nuclear accumulation of p53 was caused by a G- >A transition in codon 244 (Gly-->Asp). In the present series, no mutations within the coding region of PTEN were found. Pten expression was observed in two autopsy cases (25%) and seven biopsy samples (70%). These data suggest that TP53 is affected in some cases, but other yet-unidentified genetic alterations might contribute to tumorigenesis in GC. Furthermore, although GC might be a monoclonal process, the presence of different tumor clones cannot be ruled out. PMID- 12734659 TI - Clinical, morphological and immunological evaluation of six patients with dysferlin deficiency. AB - Limb girdle muscular dystrophy (LGMD) type 2B and distal Miyoshi myopathy (MM) are caused by mutations in a recently discovered mammalian gene coding for a skeletal muscle protein called dysferlin. The protein is normally expressed at the skeletal muscle level and absent or reduced in affected patients. We selected a clinically heterogeneous population of Italian myopathic patients with clinical evidence of myopathy and/or hyperCKemia, EMG myopathic pattern, and no alterations of the dystrophin-sarcoglycan complex. Calpain, merosin, emerin and caveolin were also tested and found normal in all patients. Dysferlin immunohistochemical and Western blot analyses allowed us to identify six patients with dysferlin deficiency: one with distal myopathy, four with limb girdle myopathy and one with hyperCKemia. No apoptosis was found in any of the six muscle specimens, although expression of the pro-apoptotic Fas antigen was mildly increased in two cases. Inflammatory reactions were present in two of the six cases, but we found no evidence of immune-mediated processes. PMID- 12734660 TI - Neuronal intranuclear inclusions distinguish familial FTD-MND type from sporadic cases. AB - Ubiquitin-immunoreactive (ub-ir) neuronal cytoplasmic inclusions are characteristically found in the extramotor cortex in patients with motor neuron disease and dementia (MND-dementia) and a subset of patients with frontotemporal dementia without motor symptoms (FTD-MND type). Recently, ub-ir neuronal intranuclear inclusions have been described in a small number of patients with familial FTD-MND type. To better define the sensitivity and specificity of this pathological change, we examined postmortem tissue from 14 patients with FTD-MND type (8 familial, 6 sporadic), 10 cases of MND-dementia (5 familial, 5 sporadic), and 19 cases of MND with no history of cognitive dysfunction (2 familial, 17 sporadic). Numerous intranuclear inclusions were found in multiple anatomic sites in 6/8 cases of familial FTD-MND. Rare intranuclear inclusions were present in the hippocampal dentate granule cells in 1 case of familial MND-dementia. No sporadic cases had intranuclear inclusions. These findings suggest that intranuclear inclusions are specific for familial FTD and may identify a subset of families with a common molecular pathogenesis. Although intranuclear inclusions are most characteristic of families in which the clinical presentation is pure FTD, they may also be found in some pedigrees with both FTD and MND; further supporting the hypothesis that FTD-MND type and MND-dementia represent a clinicopathological spectrum of disease. PMID- 12734661 TI - Neuronal expression of alphaB crystallin in cerebral infarction. AB - alphaB crystallin (alphaBC) is one of the heat shock proteins that are induced under stressful conditions. In normal brains, alphaBC is present in oligodendrocytes and astrocytes, but not in neurons. Neuronal alphaBC expression in the central nervous system under pathological conditions has been investigated in several previous studies, most of which dealt with various neurodegenerative diseases with and without ballooned neurons. Neuronal expression of alphaBC has seldom been studied in cerebral infarction (CI), and the frequency of alphaBC positive neurons in the various stages of CI is unknown. To investigate this issue, we examined 48 autopsy brains of patients with CI, and found neuronal expression of alphaBC in 68.8% of the cases. We found three types of alphaBC positive neurons: normal morphological, convex, and ballooned neurons. Although alphaBC-positive neurons were present in the every stage of CI, they were more frequent later than 10 days after the onset of CI, and the frequency of alphaBC positive ballooned neurons was particularly increased in the later stages of CI. This may indicate that morphologically normal neurons gradually swelled up through convex neurons, finally forming ballooned neurons. Previous studies indicated that alphaBC might have a cytoprotective function as a molecular chaperone, and we speculate that alphaBC is expressed in neurons subjected to ischemic stress, and exerts a cytoprotective effect on the neurons. PMID- 12734662 TI - Galanin and galanin receptors in human gliomas. AB - Galanin-like immunoreactivity (GAL-LI) and specific GAL binding sites have been shown to be widely distributed in the central nervous system (CNS) and in CNS tumors. GAL and its receptors have also been shown to be present in glial cells, but to date it is still unknown whether human gliomas produce GAL and express GAL receptors. In this study 20 brain tumors consisting of 15 glioblastomas, 4 meningiomas and 1 gliosarcoma were investigated for the presence of GAL-LI and GAL receptors. Immunofluorescence analysis revealed a dense network of GAL-LI positive cellular processes and cell bodies in 18 of the 20 tumors. In contrast, in vitro (125)I-labeled GAL receptor autoradiography showed substantial GAL binding in only 6 glioblastoma tissues. Reverse transcription-PCR analysis detected mRNA of all three known galanin receptors in the tumor tissues, with most tumors expressing multiple receptor subtypes. Pharmacological analysis of tumor membrane homogenates with GAL and the specific GAL receptor GalR2 agonist, AR-M1896, revealed that the GAL receptor GalR1 is most likely the receptor responsible for the observed GAL binding in the glioblastomas. No correlation could be found between GAL-LI, the level of GAL binding and proliferative activity as determined by immunostaining with the cell proliferation marker Ki 67. PMID- 12734663 TI - Cajal-Retzius cells, inhibitory interneuronal populations and neuropeptide Y expression in focal cortical dysplasia and microdysgenesis. AB - Focal cortical dysplasia (FCD) and microdysgenesis (MD) are likely to represent abnormalities of radial neuronal migration during cortical development. We investigated the distribution of reelin-positive Cajal-Retzius cells, known to be important in the later stages of radial neuronal migration and cortical organization, in 12 surgical cases of both MD and FCD. Quantitation revealed significantly higher numbers of these cells in MD cases compared to controls. As the majority of cortical interneurones arise via tangential rather than radial migration, we studied the distribution and morphology of inhibitory interneuronal subsets immunolabelled for calbindin, parvalbumin and calretinin within these malformations. Frequent findings were a reduction of inhibitory interneurones in the region of FCD and abnormally localised hypertrophic or multipolar calbindin positive interneurones in both FCD and MD. Neuropeptide Y immunostaining showed a striking increase in the density of the superficial plexus of fibres in both MD and FCD cases in addition to labelling of dysplastic neurones, which may represent an adaptive anti-convulsant mechanism to dampen down seizure propagation. PMID- 12734664 TI - Oxidative stress and predominant Abeta42(43) deposition in myopathies with rimmed vacuoles. AB - This study was undertaken to determine the C terminus of amyloid beta protein (Abeta), accumulated in vacuolated muscle fibers, and compare these findings to the level of oxidative stress. Eight patients with myopathies characterized by rimmed vacuoles (RVs) were analyzed. Monoclonal antibodies specific to Abeta40 or Abeta42(43) revealed that the Abeta42(43) immunoreactivity was solely distributed in the vacuolated muscle fibers, and that only a part was also immunopositive for anti-Abeta40. Quantitative analyses in four specimens, in which eight or more vacuolated muscle fibers were observed, revealed that the mean incidence of Abeta42(43)-positive muscle fibers was 79.5+/-6.2% in total vacuolated muscle fibers, whereas that of the Abeta40-positive fibers was 42.9+/-12.6%. The predominance of Abeta42(43) deposition was statistically significant ( P<0.05). Abeta deposition was then compared with the distribution of oxidative nucleic acid damage in muscle fibers using a monoclonal antibody against 8-hydroxy-2' deoxyguanosine and 8-hydroxyguanosine (8OHdG&G). The cytoplasmic staining for anti-8OHdG&G was found not only in vacuolated muscle fibers, but also in other muscle fibers including morphologically normal ones. Positive staining was completely abolished by RNase pretreatment and, thus, was suggested to reflect an increase of cellular RNA oxidation. The distribution of 8OHdG&G was much broader than the Abeta deposition. These data suggest that Abeta42(43) is predominantly involved in the pathogenesis of muscle fiber degeneration with RVs, and that oxidative damage may precede Abeta deposition in muscle fibers and play a key role in the pathomechanism of myopathies with RVs. PMID- 12734666 TI - Cytokines in sural nerve biopsies from inflammatory and non-inflammatory neuropathies. AB - Proinflammatory cytokines are supposed to play a major role in the pathophysiology of vasculitis and in the development of neuropathic pain. Here we studied the cytokine expression in sural nerve biopsy specimens from patients with vasculitic and other inflammatory and non-inflammatory neuropathies, and investigated whether an increased cytokine expression was correlated with the presence of neuropathic pain. We used immunohistochemistry including double labeling and morphometry to localize and quantify the expression of interleukin-1 beta (IL-1beta), IL-6, and tumor necrosis factor-alpha (TNF) in sural nerve biopsy samples of 41 patients with vasculitic neuropathy (VANP), chronic inflammatory demyelinating neuropathy (CIDP), non-inflammatory chronic axonal neuropathy (CANP), and 3 controls. Overall cytokine immunoreactivity was highest in VANP, less strong in CIDP and lowest in CANP. Cytokine immunoreactivity was directly correlated with the degree of axonal degeneration, endoneurial macrophages and epineurial T cells. In VANP and CANP, a higher cytokine content was associated with neuropathic pain. PMID- 12734665 TI - Localization of claudin-3 in tight junctions of the blood-brain barrier is selectively lost during experimental autoimmune encephalomyelitis and human glioblastoma multiforme. AB - In the central nervous system (CNS) complex endothelial tight junctions (TJs) form a restrictive paracellular diffusion barrier, the blood-brain barrier (BBB). During inflammation, BBB properties are frequently lost, resulting in brain edema. To investigate whether BBB leakiness correlates with molecular changes at BBB TJs, we performed immunofluorescence stainings for TJ molecules in a mouse model of experimental autoimmune encephalomyelitis (EAE) and in human tissue with glioblastoma multiforme (GBM). In TJs of healthy CNS vessels in both mouse and man we detected occludin, ZO-1, claudin-5 and claudin-3. In EAE brain and spinal cord sections we observed the selective loss of claudin-3 immunostaining from TJs of venules surrounded by inflammatory cuffs, whereas the localization of the other TJ proteins remained unchanged. In addition, selective loss of claudin-3 immunostaining was also observed in altered cerebral microvessels of human GBM. Our data demonstrate the selective loss of claudin-3 from BBB TJs under pathological conditions such as EAE or GBM when the integrity of the BBB is compromised, and therefore suggest that claudin-3 is a central component determining the integrity of BBB TJs in vivo. PMID- 12734667 TI - Primary lateral sclerosis: a rare upper-motor-predominant form of amyotrophic lateral sclerosis often accompanied by frontotemporal lobar degeneration with ubiquitinated neuronal inclusions? Report of an autopsy case and a review of the literature. AB - We report the autopsy findings of an 82-year-old woman who exhibited slowly progressive upper motor neuron signs (pseudobulbar palsy, muscle weakness and positive Babinski's sign) in the absence of lower motor neuron signs, which were followed by progressive dementia and frontotemporal atrophy, and who died 7 years and 4 months after onset of the disease. In this patient, the upper motor neuron system, including the precentral cortex and descending pyramidal tract, was severely degenerated, but the lower motor neurons and innervated skeletal muscles were well preserved. A few lower motor neurons were found to contain cytoplasmic inclusion bodies characteristic of amyotrophic lateral sclerosis (i.e., Bunina bodies and ubiquitin-positive skeins). However, fragmentation of the Golgi apparatus was not evident in the anterior horn cells examined. Therefore, it was considered that the lower motor neurons were also involved, but that the rate of degeneration of these neurons was very slow in the disease process. Marked frontotemporal lobar degeneration characterized by microvacuolation, and ubiquitin-positive neuronal inclusions and dystrophic neurites in cortical layer II were also observed, the precentral cortex being the most severely affected area. Similar ubiquitin-positive structures were also observed in the neostriatum. Finally, a survey of the literature based on this patient's clinical and pathological features led us to conclude that the rare clinical syndrome of primary lateral sclerosis is, in general, a rare upper-motor-predominant form of amyotrophic lateral sclerosis that is often accompanied by frontotemporal lobar degeneration with ubiquitinated neuronal inclusions. PMID- 12734669 TI - Comments on Vorwerk: non-traumatic vascular emergencies: management of occluded hemodialysis shunts and venous access. PMID- 12734670 TI - MR imaging-guided laser ablation of osteoid osteomas with use of optical instrument guidance at 0.23 T. AB - The purpose of this study was to determine the feasibility and features of low field MR imaging in performing interstitial laser ablation of osteoid osteomas. Between September 2001 and April 2002, five consecutive patients with clinical and imaging findings suggesting osteoid osteoma and referred for removal of osteoid osteoma were treated with interstitial laser treatment. A low-field open configuration MRI scanner (0.23 T, Outlook Proview, Philips Medical Systems, Finland) with optical instrument guidance hardware and software was used. Laser device used was of ND-Yag type (Fibertom medilas, Dornier Medizin Technik, Germany). A bare laser fiber (Dornier Medizin Technik, Germany) with a diameter of 400 microm was used. Completely balanced steady-state (CBASS; true fast imaging with steady precession) imaging was used for lesion localization, instrument guidance, and thermal monitoring. A 14-G (Cook Medical, USA) bone biopsy drill was used for initial approach. Laser treatment was conducted through the biopsy canal. All the lesions were successfully localized, targeted, and treated under MRI guidance. All the patients were symptom free 3 weeks and 3 months after the treatment. There was one recurrence reported during follow-up (6 months). The MRI-guided percutaneous interstitial laser ablation of osteoid osteomas seems to be a feasible treatment mode. PMID- 12734671 TI - High-resolution T1-weighted 3D real IR imaging of the temporal bone using triple dose contrast material. AB - The small structures in the temporal bone are surrounded by bone and air. The objectives of this study were (a) to compare contrast-enhanced T1-weighted images acquired by fast spin-echo-based three-dimensional real inversion recovery (3D rIR) against those acquired by gradient echo-based 3D SPGR in the visualization of the enhancement of small structures in the temporal bone, and (b) to determine whether either 3D rIR or 3D SPGR is useful for visualizing enhancement of the cochlear lymph fluid. Seven healthy men (age range 27-46 years) volunteered to participate in this study. All MR imaging was performed using a dedicated bilateral quadrature surface phased-array coil for temporal bone imaging at 1.5 T (Visart EX, Toshiba, Tokyo, Japan). The 3D rIR images (TR/TE/TI: 1800 ms/10 ms/500 ms) and flow-compensated 3D SPGR images (TR/TE/FA: 23 ms/10 ms/25 degrees) were obtained with a reconstructed voxel size of 0.6 x 0.7 x 0.8 mm3. Images were acquired before and 1, 90, 180, and 270 min after the administration of triple dose Gd-DTPA-BMA (0.3 mmol/kg). In post-contrast MR images, the degree of enhancement of the cochlear aqueduct, endolymphatic sac, subarcuate artery, geniculate ganglion of the facial nerve, and cochlear lymph fluid space was assessed by two radiologists. The degree of enhancement was scored as follows: 0 (no enhancement); 1 (slight enhancement); 2 (intermediate between 1 and 3); and 3 (enhancement similar to that of vessels). Enhancement scores for the endolymphatic sac, subarcuate artery, and geniculate ganglion were higher in 3D rIR than in 3D SPGR. Washout of enhancement in the endolymphatic sac appeared to be delayed compared with that in the subarcuate artery, suggesting that the enhancement in the endolymphatic sac may have been due in part to non-vascular tissue enhancement. Enhancement of the cochlear lymph space was not observed in any of the subjects in 3D rIR and 3D SPGR. The 3D rIR sequence may be more sensitive than the 3D SPGR sequence in visualizing the enhancement of small structures in the temporal bone; however, enhancement of the cochlear fluid space could not be visualized even with 3D rIR, triple-dose contrast, and dedicated coils at 1.5 T. PMID- 12734672 TI - Paraneoplastic eosinophilic fasciitis: a case report. AB - A 60-year-old white woman with polycythemia rubra vera post splenectomy in November 2001 was found to have peripheral white blood cell counts increasing over 3 months. Cytogenetics revealed trisomy of chromosomes 8 and 9, and bone marrow biopsy showed hypercellular, fibrotic bone marrow consistent with myelofibrosis of polycythemia rubra vera. Two months later, the patient developed acute swelling and pain in her lower extremities. The clinical symptoms along with confirmatory histology supported the diagnosis of eosinophilic fasciitis. This is the first reported case in the English literature of an association between polycythemia vera and eosinophilic fasciitis. PMID- 12734674 TI - An orally active Amazonian plant extract (BIRM) inhibits prostate cancer growth and metastasis. AB - PURPOSE: Poor efficacy of conventional chemotherapeutic drugs against metastatic hormone-refractory prostate cancer (CaP) drives patients to try "alternative medicine". The antitumor activity of one such agent, "BIRM" (biological immune response modulator; "Simple Ecuadorian Oral Solution: an extract of an Amazonian plant"), was characterized in vitro and in vivo using established CaP cell lines and a tumor model. METHODS: The cytotoxicity of BIRM in four human and one rat CaP cell line was evaluated using cell proliferation-inhibition and clonogenic survival assays. BIRM-induced apoptosis, alterations in cell cycle phase progression and inhibition of the extracellular matrix-degrading enzyme hyaluronidase were also investigated in these cells. The in vivo efficacy of BIRM was evaluated in rats with subcutaneous tumor implants of Dunning EGFP-MAT LyLu cells. The active species in BIRM were characterized by gel filtration chromatography. RESULTS: BIRM inhibited cell proliferation and clonogenic growth of the CaP cells (IC(50) about 8.0 microl/ml). It increased cell accumulation in the G(0)/G(1) phase by 33.8% and decreased the proportion of cells in S phase by 54.6%. Apoptotic cell death in BIRM-treated cells was associated with activation of cell death-associated caspases. BIRM inhibited the activity of hyaluronidase, a hyaluronic acid-degrading enzyme, at 1 microl/ml. Treatment of MAT LyLu tumor bearing rats with BIRM by oral gavage resulted in a significant decrease in tumor incidence (50%), tumor growth rate (18.6+/-1.3 days for 1 cc tumor growth in control rats and 25.7+/-2.6 days in BIRM-treated rats), and only one out of six BIRM-treated rats versus four out of six in the control group developed lung metastasis. Three active ingredients in BIRM with a relative molecular mass (Mr) of >or=3500 were identified by ultracentrifugation and gel filtration chromatography and were found to be resistant to proteinase and heat (100 degrees C). CONCLUSION: The plant extract BIRM contains antitumor compounds of Mr >or=3500 with potent antiproliferative activity in vitro and in vivo against prostate cancer cells. PMID- 12734675 TI - Prolonged treatment with imatinib mesylate in patients with advanced chronic myeloid leukemia causes a reduction of bcr/abl mRNA levels independent of cytogenetic response. AB - Bcr/abl mRNA levels were monitored in 13 patients with chronic myeloid leukemia receiving imatinib mesylate over a period of 78 weeks. During treatment median bcr/abl mRNA levels progressively declined from 77.2 normalized dose (nD) at baseline to 11.28 nD after 13 weeks ( P<0.05) and to 1.28 nD after 78 weeks ( P<0.05). After 13 weeks, bcr/abl mRNA levels were significantly lower in cytogenetic responders compared to nonresponders ( P<0.05), but subsequent decrease in the transcript levels caused the loss of any correlation to the cytogenetic status. These results suggest that bcr/abl mRNA levels may reflect cytogenetic response only during the early phases of imatinib therapy. PMID- 12734673 TI - Rsf1p, a protein required for respiratory growth of Saccharomyces cerevisiae. AB - A central problem in our understanding of mitochondrial (mt) function remains the question of how coordinate transcriptional control is accomplished between nucleus and mitochondria. Here, we report the initial characterization of a protein of previously unknown function, the product of the YMR030 W gene, that appears to mediate such coordinate gene expression. Expression of YMR030 W is glucose-repressible; a deletion mutant for this gene shows a severe growth defect on glycerol-, but not glucose- or ethanol-based medium. In that mutant, transcript levels from GUT1 and GUT2 are highly attenuated compared with those of the wild-type parent when both are grown on glycerol-based medium. Under the same growth conditions, transcripts from the mt OLI1 gene, which has one copy of a mt upstream activating sequence (UAS) in its 5'-flanking region, are attenuated in the DeltaYMR030 W mutant, but mRNA from the mt COX3 ( OXI2) gene, which lacks the mt UAS, are not. Some nuclear genes encoding mt-related proteins also show low transcript levels in the DeltaYMR030 W mutant in comparison with those of the wild-type parent strain during glycerol-based growth. Localization of the protein, via its expression fused to green fluorescent protein, indicates that it is present in both nucleus and mitochondria, supporting a respiration-related transcriptional role for this gene product in both cellular genetic compartments. Because of its role in both respiratory growth and mt function, we designate the YMR030 W coding sequence RSF1 (respiration factor 1). PMID- 12734676 TI - Assessment of the 20210 G to A prothrombin variant in a sample of patients from the French Basque Country with various thrombophilic conditions. AB - We investigated the distribution of the prothrombin variant G20210A (PT20210A) in a sample of 103 patients (mean age: 34.5 years) living in the French Basque Country and presenting with conditions known to be significantly associated with this peculiar mutation according to a literature review. These patients suffered from repeated personal or familial venous thromboses, stroke at young age, or repetitive maternal-fetal disorders (abortions, preeclampsia, fetal growth retardation). Five patients (4.8%) were found to be carriers: two homozygotes and three heterozygotes (one also heterozygote for factor V Leiden). Of note, two presented with mesenteric venous thrombosis. The distribution of PT20210A among our sample was comparable to data from Western European series albeit a tendency for lower mutation prevalence was observed in our subgroup with obstetrical disorders. In addition, no significant difference in PT20210A frequency was evidenced between autochthonous Basques and individuals from other origins. PMID- 12734677 TI - Idiopathic thrombocytopenic purpura in pregnancy: a single institutional experience with maternal and neonatal outcomes. AB - We observed 13 pregnant women of 70 females with idiopathic thrombocytopenic purpura (ITP) from January 1992 through September 2002. Thirteen mothers with ITP gave birth to twelve babies and two fetuses died. One of the pregnancies produced twins. Seven of the cases were diagnosed with ITP before pregnancy and six during pregnancy. One of the thirteen pregnancies was complicated by preeclampsia, one by ablatio placentae, and one by intrauterine death. Seven mothers received corticosteroid treatment, four high-dose immunoglobulin therapies, and one underwent splenectomy in the second trimester of gestation. At the time of delivery six mothers had normal platelet counts and seven had low platelet counts. Nine deliveries were by vaginal route and four were by cesarean section. Eleven infants were born with normal platelet counts and one was thrombocytopenic at the time of delivery. No infant showed any clinical signs of hemorrhage and there were no neonatal complications. Two fetuses died; one of them because of ablatio placentae and the other was intrauterine dead. In conclusion, ITP in pregnancy requires the management of two patients, the mother and her baby; hence, the close collaboration of a multidisciplinary group composed of a hematologist, obstetrician, anesthesiologist, and neonatologist is essential. PMID- 12734678 TI - The intersesamoidal ridge of the first metatarsal bone: anatomical basics and clinical considerations. AB - The first metatarsal shows a ridge at the plantar side of the head, aligned nearly longitudinally. On 100 macerated first metatarsals (53 right, 47 left) the existence and alignment of this ridge was examined. On the basis of a regression formula, the specimens were assigned to the genders; accordingly, 29 bones were female, the remaining 71 bones male. The intersesamoidal ridge existed in 95 cases (50 right, 45 left); all 29 female bones showed this ridge, male ones in 66 cases. The ridge deviated on average 7.99 degrees from the longitudinal axis of the first metatarsal towards distal-lateral (from -3 degrees to 19 degrees ). Significant differences ( p=0.001) were found for side (left: 6.04 degrees, right: 9.74 degrees ) but not for gender (female: 8.14 degrees, male: 7.92 degrees ). Noticeable correlations with the length of the first metatarsal and the size of the head could not be determined. The mean values of the alignment of the intersesamoidal ridge seem also not to correlate with the metatarsophalangeal angle. The ridge separates the joint surfaces for the sesamoid bones, which are built-in to the tendons of the two heads of the flexor hallucis brevis muscle. Thus, from the alignment of the ridge, the functional direction of this muscle can be concluded. Since the alignment deviates from the longitudinal axis of the first metatarsal, an adduction, in addition to the flexion, can be concluded. Operative techniques for the correction of hallux valgus often change the alignment of the ridge, and therefore also the biomechanical properties of the flexor hallucis brevis muscle. This could explain failures in several surgical therapies. PMID- 12734679 TI - Accuracy of laparoscopy in the diagnosis and staging of lymphoproliferative diseases. AB - Laparoscopy seems to be playing an emerging role in the management of lymphoproliferative diseases. The aim of this study was to prospectively analyze personal experience evaluating the role and limits of laparoscopy in the management of lymphomas. From July 1993 to December 2000, 131 consecutive patients were referred to our institution for primary diagnosis (group A, n = 70), suspected relapse (reassessment) (group B, n = 54), or staging/restaging of lymphoproliferative diseases (group C, n = 7). Diagnostic and/or operative laparoscopy was performed in all patients. To assess the accuracy of laparoscopy, the results were analyzed according to the indications for surgery. In all, 128 procedures were completed laparoscopically (95.5%). Conversion was required in 7 cases (5.1%). Causes of the conversions were severe obesity (body mass index 62.5), uncontrolled intraoperative bleeding (HIV+), nondiagnostic tissue sampling (2 cases), perisplenic inflammation and perisplenic abscesses (3 cases). The results of this study highlight the safety of diagnostic and staging laparoscopy and laparoscopic splenectomy in patients with lymphoproliferative diseases (major complications 2.9%, perioperative mortality 0%). In all, 96.4% of patients from group A and 100% of patients from group B were treated on the basis of laparoscopic findings. No false negative diagnosis occurred. Laparoscopy may become the "gold standard" in the management of lymphoproliferative disease in the following settings: for the differential diagnosis of hepatic and/or splenic focal lesions; when percutaneous needle biopsy fails and/or genetic analysis is needed for therapeutic decision; for the primary diagnosis and abdominal staging of patients with diffuse retroperitoneal lymphadenopathy in the absence of peripheral lymphadenopathy; for cases of abdominal restaging after concurrent chemoradiotherapy and in cases of suspected relapse when percutaneous biopsy is not technically possible; and for patients with lymphoproliferative disease when splenectomy is required. Marked splenomegaly with perisplenic inflammatory reaction and lymphadenopathy in HIV+ patients should be considered possible causes of failure of the laparoscopic approach. PMID- 12734680 TI - Doxorubicin activity is enhanced by hyperthermia in a model of ex vivo vascular perfusion of human colon carcinoma. AB - There is little information on the pharmacokinetics and pharmacodynamics of doxorubicin (DXR) administered during locoregional treatments of colon carcinoma under hyperthermic conditions. The aim of this study was to evaluate distribution and activity of DXR in healthy tissue and tumor tissues under hyperthermic conditions by using an experimental model of ex vivo isolated vascular perfusion of human colon segments bearing primary carcinoma. The influence of topoisomerase II alpha (TPI2 alpha) expression on the anti-cancer activity of DXR combined with heat was evaluated as well. Twenty seven colon segments surgically resected for primary carcinoma were perfused ex vivo under physiological conditions (group A, n = 7), with DXR (group B, n = 6), under hyperthermic conditions (group C, n = 6), and with the combination DXR-hyperthermia (group D, n = 8). Samples of perfusate and tissues (normal and pathologic) were collected during 90 minutes of perfusion. Doxorubicin concentration in perfusate and tissues was assessed with high-performance liquid chromatography. Protein expression of TPI2 alpha, the main molecular target of DXR, was measured by image analysis in normal mucosa and tumor samples. Drug uptake was increased by heat equally in healthy and diseased tissue. Under hyperthermic conditions, DXR activity was significantly higher in pathologic mucosa than in normal mucosa. Furthermore, the activity of DXR combined with heat correlated with baseline TPI2 alpha levels in tumor tissue. From these findings, it appears that heat sensitizes tumor cells-but not normal mucosa-to DXR activity. Furthermore, protein levels of TPI2 alpha in pretreatment samples could predict tumor sensitivity to DXR. PMID- 12734681 TI - Persistent primary hyperparathyroidism caused by adenomas identified in pharyngeal or adjacent structures. AB - Abnormalities in the normal migration of the parathyroid glands during embryological development of the head and neck may result in considerable variability in the location of parathyroid tissue. Ectopic parathyroid adenomas present diagnostic and technical challenges and are frequently the cause of persistent primary hyperparathyroidism (HPT) after unsuccessful initial parathyroid surgery. We report a series of eight patients with persistent primary HPT who had adenomas in rare and unusual locations associated with various pharyngeal structures. Four were located within the epineurium of the vagus nerve at or above the level of the carotid bifurcation, and four were located within the paranasopharyngeal space or oropharynx. Noninvasive and invasive preoperative imaging studies were crucial in localizing the neoplasms in these patients and permitted the use of a direct surgical approach, resulting in cure in all patients and a low complication rate. The location of parathyroid glands in high pharyngeal and cervical structures is a consequence of anomalous or arrested descent through developing pharyngeal structures and illustrates the remarkable spectrum of ectopic parathyroid adenomas that occur secondary to this phenomenon. PMID- 12734682 TI - Physiologic studies on nitric oxide in rat small bowel isografts. AB - The enteric nervous system (ENS), especially the nonadrenergic noncholinergic (NANC) inhibitory nerves, is an important factor in intestinal peristalsis. Recently, it was established that nitric oxide (NO) is released after stimulation of NANC inhibitory nerves. Inhibitory nerves such as NANC inhibitory nerves in the ENS are more easily damaged than excitatory nerves by reperfusion or ischemic injuries during small bowel transplantation (SBT). To evaluate the effects of reperfusion and ischemic injuries to the ENS in the transplanted small bowel, we examined the ENS responses, including the effects of NO in the isografted rat jejunum, using the nontransplanted jejunum as a control. To avoid potentially confounding immune phenomena, we used syngeneic Lewis (LEW) rats. Orthotopic entire SBT with portocaval drainage was performed from LEW rats to LEW rats. Isografted muscle strips were obtained from 8 LEW rats 130 days after SBT (n = 24). As controls, normal muscle strips of the jejunum were obtained from 20 nontransplanted LEW rats (n = 60). A mechanograph was used to evaluate in vitro jejunal responses to electrical field stimulation of the adrenergic and cholinergic nerves before and after treatment with various autonomic nerve blockers, N(G)-nitro-Ll-arginine ( L-NNA), and L-arginine. The results indicated that excitatory nerves, especially NANC excitatory nerves, were more dominant in the isografted jejunum than in the normal jejunum (p < 0.01). NANC inhibitory nerves were found to act on the normal jejunum and to a lesser extent on the isografted jejunum (p < 0.05). NO mediates the relaxation reaction of NANC inhibitory nerves in the normal jejunum and to a lesser extent in the isografted jejunum. These results indicated that the intrinsic intestinal innervation contains excitatory and inhibitory nerves and that the former, especially NANC excitatory nerves, are more dominant in the isografted jejunum than in the normal jejunum. In addition, reduction of the action of NANC inhibitory nerves such as that by NO may be largely related to impaired motility in the isografted jejunum. Thus over a long period of time (more than 130 days after SBT) transplanted small bowel dysmotility may be influenced by reperfusion or ischemic injury to the ENS (especially NANC inhibitory nerves) via NO in the transplanted jejunum after syngeneic SBT. PMID- 12734683 TI - Pathophysiological studies on the relationship between postgastrectomy syndrome and gastric emptying function at 5 years after pylorus-preserving distal gastrectomy for early gastric cancer. AB - Pylorus-preserving distal gastrectomy (PPG) has frequently been performed on patients with early gastric cancer in Japan to prevent the postgastrectomy syndrome seen after conventional distal gastrectomy (CDG). The long-term postoperative quality of life (QOL) and gastric emptying function in patients after PPG has not been assessed in detail. To clarify the usefulness of PPG for treating early gastric cancer we investigated the relation between postgastrectomy syndrome and gastric emptying function 5 years after PPG and then compared the results with those 5 years after CDG. Altogether, 32 patients who underwent curative gastrectomy at our clinic for early gastric cancer (submucosal cancer without lymph node metastasis) were studied. Ten subjects who underwent PPG with D2 lymphadenectomy without preserving the hepatic and pyloric branches of the vagal nerve [group A: eight men, two women; age 33-70 years (mean 60.7 years)] were interviewed and asked about appetite, weight loss, epigastric fullness, reflux esophagitis, and early dumping syndrome. They were compared with patients after CDG [group B: 36-72 years (mean 63.6 years)]. Esophagogastric endoscopy, abdominal ultrasonography, and gastric emptying function were also studied. The gastric emptying time of a semisolid diet was measured with a radioisotope method using (99m)Tc-labeled rice gruel; the gastric emptying time of a liquid diet was measured with the acetaminophen method using orange juice. The control subjects (group C) consisted of 18 healthy volunteers (10 men, 8 women) without gastrointestinal symptoms aged 38 to 68 years (mean 60.8 years). The following results were obtained: PPG (group A) alleviated postoperative gastrointestinal symptoms such as appetite loss, reflux esophagitis, early dumping syndrome, lost body weight, endoscopic reflux esophagitis, endoscopic gastritis in the remnant stomach, and postogastrectomy cholecystolithiasis better than did CDG (group B). The only weak point with the PPG procedure was that it produced a feeling of epigastric fullness. The pattern of the gastric emptying curve for the semisolid diet was almost the same among groups A, B, and C, although delayed gastric emptying was clearly more frequent in group A than in group B or C ( p < 0.05). Gastric emptying with the liquid diet in group B was significantly faster than that in groups A and C ( p < 0.01). Gastric emptying in groups A and C was similar. These results showed that PPG improved the postoperative QOL, but the delayed emptying of semisolid diet after PPG led to a feeling of epigastric fullness after meals due to retention of contents in the residual stomach. Epigastric fullness after meals continued in many patients after PPG. Thus the only disadvantage of the PPG procedure is the sensation of epigastric fullness and gastric stasis due to delayed gastric emptying of a semisolid diet. PMID- 12734684 TI - Implant-related complications in the treatment of unstable intertrochanteric fractures: meta-analysis of dynamic screw-plate versus dynamic screw intramedullary nail devices. AB - The choice between dynamic screw-intramedullary nail (DSIN) devices and dynamic screw-plate (DSP) devices for the fixation of unstable trochanteric fractures remains controversial. This study presents a meta-analysis of fixation failures in unstable trochanteric femoral fractures using DSP devices or DSIN devices. Two independent assessors selected randomised controlled trials using a range of electronic databases, as well as reference lists of selected articles. A study quality checklist was used. The occurrence of fixation failure, in particular cut out, was the primary subject of analysis using descriptive statistics and random effect meta-analyses. Seventeen trials were identified. Meta-analyses showed no significant difference in the frequency of implant-related complications between the two types of devices. Iatrogenic femoral fractures associated with the use of DSIN devices represent a rare, but persistent, risk. There was a tendency for less frequent cut-out with intramedullary devices compared with DSP devices. PMID- 12734686 TI - Gallium-67 activated charcoal: a new method for preparation of radioactive capsules for colonic transit study. AB - Indium-111 is currently the radionuclide of choice for colonic transit study. However, it is expensive and not available in many hospitals. Technetium-99m has been proposed for colonic transit study but the short half-life has limited its use. Gallium-67 citrate is inexpensive and available in most countries. Most importantly, it has a suitable half-life for colonic transit study. Attempts have been made in some studies to use (67)Ga citrate to label activated charcoal, but the results have not been good because of poor stability. In this study, we successfully labelled activated charcoal()with (67)Ga citrate by adding alcohol and 5% glucose solution. To evaluate the in vitro stability, the (67)Ga-activated charcoal was incubated in a milieu mimicking the intestinal content, containing lipase, trypsin and glycochenodeoxycholate at different pH values (6.0, 7.0, 7.4 and 8.0) and for different durations (0 h, 24 h, 48 h, 72 h and 96 h). For the in vivo study, the (67)Ga-activated charcoal was loaded into a commercial empty enteric capsule. Colonic transit scintigraphy was performed in five volunteers, including three healthy people and two constipated patients, after intake of the radioactive capsule. Images were obtained at 2 h, 4 h, 6 h, 8 h, 24h, 48 h, 72 h etc. until no radioactivity was detected in the bowel. Our data show that the in vitro stability of (67)Ga-activated charcoal was good. The labelling efficiency still exceeded 91% at 96 h at pH values of 6.0, 7.0 and 7.4. In the group with a pH value of 8.0, the labelling efficiency gradually fell during the 4-day incubation but was still higher than 88% at the end of the fourth day. In the in vivo study, most capsules disintegrated in the caecum/colon region, and the (67)Ga-activated charcoal mixed very well with bowel content. In addition, the radioactive charcoal could be detected clearly on the 72-h image, which is very important for the evaluation of colonic transit time in patients with constipation. In conclusion, activated charcoal labelled with (67)Ga citrate is a potential radioactive marker for colonic transit study. PMID- 12734687 TI - Epidemiology of Hodgkin's disease and non-Hodgkin's lymphoma. AB - This review considers in detail the descriptive and aetiological epidemiology of Hodgkin's disease and non-Hodgkin's lymphoma (NHL), with attention to histological subcategories when the literature allows. The aetiology of Hodgkin's disease remains only partially understood. There is evidence that Epstein-Barr virus (EBV) may be involved in the causation of some cases, and clinical immune deficiency is a risk factor for a few, but the evidence is not entirely consistent and other factors may also be important in causing the EBV-associated cases of Hodgkin's disease. The cause of EBV-unassociated cases remains obscure. For NHL, although it has been shown that some cases are related to immune deficiency and chronic antigenic stimulation, and especially to EBV in the context of immune deficiency, the causation of the majority of cases remains unknown. The increasing incidence of NHL, other than that related to AIDS, is also essentially unexplained. Epidemiological investigation of the aetiology of NHL and Hodgkin's disease is making steady progress, however, and there remain leads to be followed that may result in a better understanding and hence prevention. PMID- 12734688 TI - The classification of lymphomas: a new beginning or the end of an era? AB - The World Health Organisation Classification of Tumours of the Haematopoietic and Lymphoid Tissues has recently been published. This is the latest in a long line of classifications of haematological malignancies and will be the international standard. It is now possible to achieve high levels of diagnostic accuracy for the main types of lymphoma. However, many of the entities encompass a wide spectrum of clinical outcomes and this approach to classification may be insufficiently precise for the future needs of haematological oncology. Rapid progress in targeted therapies may require further developments in tumour classification based on pathogenic features rather than arbitrary morphological criteria. PMID- 12734689 TI - Functioning pulmonary metastases of thyroid cancer: does radioiodine influence the prognosis? AB - Functioning pulmonary metastases are the most common distant lesions of differentiated thyroid cancer. About 50% of patients with such metastases die within 10 years. The impact of iodine-131 therapy is controversial. In this study we examined: (1) the early diagnostic value of post-surgery (131)I ablation for lung invasion and (2) the survival of patients receiving periodic (131)I therapy. Between January 1970 and December 1995 we provided initial treatment for 509 patients with thyroid cancer. Most of them (74%) underwent total thyroidectomy and (131)I ablation. Functioning pulmonary metastases occurred in 20 patients. All these patients received periodic (131)I therapy for as long as (131)I uptake persisted. Additional therapy consisted of lung surgery in three patients and local treatment of bone lesions in four patients. Follow-up data were recorded up to December 2001. Functioning pulmonary metastases occurred late in one patient, and were visible on the post-surgery (131)I therapy scan in the other 19 patients. At diagnosis of lung invasion, 11 patients had negative chest X-ray findings, and serum thyroglobulin levels were not suggestive of metastatic disease in 56% of these cases. One of the 11 patients with negative chest X-ray findings died with a neck recurrence, two have persistent pulmonary (131)I uptake, and the other eight are in apparent remission after receiving an average cumulative (131)I activity of 338 mCi (12.51 GBq). The nine patients with positive chest X-ray findings received an average of 939 mCi (34.74 GBq); two of them died, five are continuing to receive therapy and two are in apparent remission. Overall survival at 10 years is 84%. The average follow-up of the 17 survivors is 12.7 years. These results suggest that patients with functioning pulmonary metastases, even in advanced stages, may survive for many years on (131)I therapy. Early diagnosis, during post-surgery (131)I scanning, of radiologically inapparent metastases is associated with a better prognosis. PMID- 12734690 TI - Validation of the dual-table autoradiographic method to quantify two sequential rCBFs in a single SPET session with N-isopropyl-[123I] p-iodoamphetamine. AB - We evaluated an autoradiographic (ARG) method to calculate regional cerebral blood flow (rCBF) sequentially before and after an acetazolamide (ACZ) challenge in a single session of single-photon emission tomography (SPET) with two injections of N-isopropyl-[(123)I] p-iodoamphetamine (IMP). The method uses a table look-up method with a fixed distribution volume (Vd) and a standard input function of IMP. To calculate rCBF after an ACZ challenge, two look-up tables (a dual-table) are used to reflect the effect of radioactivity in the brain from the first dose of IMP. We performed simulation studies to evaluate errors attributable to (a) a change in rCBF induced by an ACZ challenge during the scan and (b) a fixed Vd value that might be different from an individual one, along with the effect of (c) scan length. Thirty-three patients were studied by dynamic SPET with two injections of IMP and frequent arterial blood sampling, and the data were analysed using the dual-table ARG method. Twenty-four of the 33 patients received an injection of ACZ 10 min before the second dose of IMP. We generated a standard input function by averaging individual input functions. The optimal method to calibrate a standard input function was determined so that the SD of differences between rCBF calculated by using a calibrated standard input function (F(SIF)) and that calculated by using an individual input function (F(IIF)) was minimised. Reliability of the method was evaluated by comparing F(SIF) with gold standard rCBF (F(REF)) obtained by two-compartment model analysis of dynamic SPET data and an individual input function with a non-linear least squares fitting method. Errors caused by (a) were less than 4% for a first rCBF ranging between 20 and 60 ml 100 g(-1) min(-1) and an rCBF change of between -25% and 50%. Errors caused by (b) were relatively large compared with those caused by (a), and were affected by (c) with an increasing error in a longer scan. In the patient study with a proposed scan protocol of 25 min for the first and 15 min for the second measurement, the error attributable to the standard input function was smaller when calibrated with a continuously drawn arterial blood sample (random error of 3.8% for continuous 10-min arterial blood sampling after the second dose of IMP) than with a single arterial blood sample (random error of 9.0% at 5 min after the second dose of IMP). Systematic and random errors of F(SIF) compared with F(REF) were 0.0% and 6.3%, respectively. The dual table ARG method can be reliably used to quantify rCBF before and after an ACZ challenge with a 40-min scan protocol and continuous arterial blood sampling for several minutes. PMID- 12734691 TI - 99mTc-MIBI scintigraphy for early detection of locally recurrent non-small cell lung cancer treated with definitive radiation therapy. AB - After radiation therapy of lung cancer, a dense fibrotic shadow develops in the irradiated lung. Owing to this fibrosis, early detection of local recurrence after treatment is sometimes difficult even when using computed tomography (CT) and magnetic resonance imaging. We investigated the diagnostic accuracy of technetium-99m hexakis 2-methoxyisobutylisonitrile ((99m)Tc-MIBI) scintigraphy for the detection of recurrent lung cancer following definitive radiation therapy. Eighteen patients with primary non-small cell lung cancer treated with radiation therapy 1 year previously were studied with (99m)Tc-MIBI scintigraphy. They showed no evidence of local recurrence on serial chest radiographs. All single-photon emission tomography (SPET) images acquired 2 h after intravenous administration of the radiopharmaceutical were visually interpreted with knowledge of the pretreatment chest radiograph, CT and the details of radiation therapy (radiation portals and administered doses). A region of interest (ROI) analysis was also performed. In addition to the ROI ratio of tumour uptake to accumulation in contralateral normal lung (tumour/lung ratio), another semiquantitative analysis, the ratio of tumour uptake to accumulation in radiation fibrosis (tumour/fibrosis ratio), was performed to differentiate between accumulation in radiation fibrosis and the tumour uptake. The scintigraphic diagnoses were correlated with clinical outcome. The sensitivity, specificity and negative predictive value of (99m)Tc-MIBI scintigraphy for the detection of recurrent lung cancer were all 88.9% (8/9). The tumour/lung ratios (mean+/-SEM) of the nine patients with local recurrence and the other eight without local failure were 2.00+/-0.11 and 1.40+/-0.09, respectively ( P<0.01). The tumour/fibrosis ratios of the patients with and those without recurrence were 1.47+/-0.08 and 0.93+/-0.05, respectively ( P<0.01). These results suggest that (99m)Tc-MIBI scintigraphy might be of value for the detection of recurrent lung cancer, and especially of small foci in areas of radiation fibrosis that are hardly noticeable on serial chest radiographs. PMID- 12734693 TI - Microbial iron respiration: impacts on corrosion processes. AB - In this review, we focus on how biofilms comprising iron-respiring bacteria influence steel corrosion. Specifically, we discuss how biofilm growth can affect the chemistry of the environment around the steel at different stages of biofilm development, under static or dynamic fluid regimes. We suggest that a mechanistic understanding of the role of biofilm metabolic activity may facilitate corrosion control. PMID- 12734694 TI - Pilot plant for bioremediation of mercury-containing industrial wastewater. AB - Mercury is an extremely toxic pollutant that is currently being emitted mainly by low level industrial sources. It is distributed globally through the atmosphere, from where it precipitates onto the surface of the Earth, enters aquatic organisms, accumulates in fish and finally affects the health of human populations. Microbes have evolved a mechanism for mercury detoxification [mercury resistance operon ( mer)] based on intracellular reduction of Hg(2+) to non-toxic Hg(0) by the mercuric reductase enzyme and subsequent diffusional loss of Hg(0) from the cell. It was shown that Hg(0) produced by microbial detoxification can be retained quantitatively in packed bed bioreactors, in which biofilms of mercury-resistant bacteria are grown on porous carrier material. This review describes operation of this system on a technical, fully automated, scale, and its operation at a chloralkali electrolysis factory. It was shown to work with high efficiency under fluctuating mercury concentrations and to be robust against transiently toxic conditions. The gradient of mercury concentration in the technical scale system exerted a strong selective pressure on the microbial community, which resulted in a succession of mercury-resistant strains at high mercury concentrations and an increase in phylogenetic and functional diversity at low mercury concentrations. Clean-up of mercury-containing wastewater by mercury-resistant microbes is a simple, environmentally friendly and cost effective alternative to current treatment technologies. PMID- 12734695 TI - Conservation of the MHC-like region throughout evolution. AB - Identification of conserved regions between the genomes of distant species is a crucial step in the reconstruction of the genomic organization of their last common ancestor. Here we confirm for the first time with robust evidence, the existence of a region of conserved synteny between the human genome and the Drosophila genome. This evolutionarily conserved synteny involves the human MHC and paralogous regions, and we identified 19 conserved genes between these two species in a Drosophila genomic region of less than 2 Mb. The statistical analysis of the distribution of these 19 genes between the Drosophila and human genomes shows that it cannot be explained by chance. Our study constitutes a first step towards the reconstruction of the genome of Urbilateria (the ancestor of all bilaterian) and allows for a better understanding of the evolutionary history of our genome as well as other metazoan genomes. PMID- 12734696 TI - High-resolution proton NMR measures mobile lipids associated with Triton resistant membrane domains in haematopoietic K562 cells lacking or expressing caveolin-1. AB - High-resolution proton NMR spectra of intact tumour cells generally exhibit intense signals due to isotropically mobile lipids (MLs) of still uncertain nature and origin. NMR studies performed on intact wild-type and caveolin-1 infected haematopoietic K562 cells showed that, under our experimental conditions, part of the ML signals are due to lipid complexes resistant to extraction in Triton X-100 at 4 degrees C. This evidence suggests that a portion of NMR-visible lipid structures are compatible with Triton-resistant membrane rafts and therefore biophysically distinct from NMR-visible Triton-soluble lipid bodies. Similarly to lipid rafts and caveolae, the organization of the Triton insoluble ML domains could be compromised by treatment with beta-octylglucoside or methyl-beta-cyclodextrin. Exposure to exogenous sphingomyelinase caused an increase in ML NMR visibility, indicating the possible involvement of ceramides in ML formation. The mobility of these lipids was found to be temperature sensitive, suggesting a transition in cells going from 4 degrees C to 25-37 degrees C. These new results are here discussed in the light of possible contributions of plasma membrane microdomains to NMR-visible ML signals. PMID- 12734697 TI - Exclusion of maltodextrins from phosphatidylcholine multilayers during dehydration: effects on membrane phase behaviour. AB - The effect of increasing solute size on phosphatidylcholine phase behaviour at a range of hydrations was investigated using differential scanning calorimetry. Dehydration of phospholipid membranes gives rise to a compressive stress within the bilayers that promotes fluid-to-gel phase transitions. According to the Hydration Forces Explanation, sugars in the intermembrane space minimize the compressive stress and limit increases in the fluid-gel transition temperature, T(m), by acting as osmotic and volumetric spacers that hinder the close approach of membranes. However, the sugars must remain between the bilayers in order to limit the rise in T(m). Large polymers are excluded from the interlamellar space during dehydration and do not limit the dehydration-induced rise in T(m). In this study, we used maltodextrins with a range of molecular weights to investigate the size-exclusion limit for polymers between phosphatidylcholine bilayers. Solutes with sizes ranging from glucose to dextran 1000 limited the rise in lipid T(m) during dehydration, suggesting that they remain between dehydrated bilayers. At the lowest hydrations the solutions vitrified, and T(m) was further depressed to about 20 degrees C below the transition temperature for the lipid in excess water, T(o). The depression of T(m) below T(o) occurs when the interlamellar solution vitrifies between fluid phase bilayers. The larger maltodextrins, dextran 5000 and 12,000, had little effect on the T(m) of the PCs at any hydration, nor did vitrification of these larger polymers affect the lipid phase behaviour. This suggests that the larger maltodextrins are excluded from the interlamellar region during dehydration. PMID- 12734698 TI - Adenophostin A and imipramine are two activators of the olfactory inositol 1,4,5 trisphosphate-gated channel in fish olfatory cilia. AB - Binding of an odorant to its receptor activates the cAMP-dependent pathway, and also leads to inositol 1,4,5-trisphosphate (InsP(3)) production. This induces opening of a plasma membrane channel in olfactory receptor cells (ORCs). We investigated single-channel properties of this channel in the presence of a phospholipase C (PLC) activator (imipramine) and of a potent activator of the InsP(3)/Ca(2+) release channel (adenophostin A) by reconstituting carp olfactory cilia into planar lipid bilayers. In the presence of 53 mM barium as a charge carrier, the addition of 50 microM imipramine induced a current of 1.53+/-0.05 pA at 0 mV. There were two different mean open times (6.0+/-0.6 ms and 49.6+/-6.4 ms). The I/ V curve displayed a slope conductance of 50+/-2 pS. Channel activity was transient and was blocked by neomycin (50 microM). These observations suggest that imipramine may activate the olfactory InsP(3)-gated channel through PLC. Using the same ionic conditions, the application of 0.5 microM adenophostin A triggered a current of 1.47+/-0.04 pA at 0 mV. The I/ V curve displayed a slope conductance of 60+/-2 pS. This channel showed only a single mean open time (15.0+/-0.3 ms) and was strongly inhibited by ruthenium red (30 microM) and heparin (10 microg/mL). These results indicate that adenophostin A and imipramine may act on the ciliary InsP(3)-gated channel and are potentially valuable pharmacological tools for studying olfactory transduction mechanisms. PMID- 12734699 TI - Inward and outward potassium currents through the same chimeric human Kv channel. AB - Voltage-gated ion channels are among the most intensely studied membrane proteins today and a variety of techniques has led to a basic mapping of functional roles onto specific regions of their structure. The architecture of the proteins appears to be modular and segments associated with voltage sensing and the pore lining have been identified. However, the means by which movement of the sensor is transduced into channel opening is still unclear. In this communication, we report on a chimeric potassium channel construct which can function in two distinct operating voltage ranges, spanning both inward and outward currents with a non-conducting intervening regime. The observed changes in operating range could be brought about by perturbing either the direction of sensor movement or the process of transducing movements of the sensor into channel opening and closing. The construct could thus provide a means to identify the machinery underlying these processes. PMID- 12734700 TI - Time-resolved fluorescence anisotropy studies show domain-specific interactions of calmodulin with IQ target sequences of myosin V. AB - Single cysteine mutants of calmodulin, Cam(S38C) and Cam(N111C), have been specifically labelled with Alexa488 maleimide to study the effects of calcium on the structural dynamics of calmodulin complexed with IQ3, IQ4 and IQ34 target peptide motifs of mouse unconventional myosin-V. Using phase fluorometry, the time-resolved anisotropy shows well-separated global and segmental correlation times. The calcium-sensitive global motion of either calmodulin domain can be independently monitored in domain-specific interactions of either apo- or Ca(4).calmodulin with IQ3 or IQ4 peptides. C-domain interactions predominate, and apo-N-domain interactions are unexpectedly weak. The 1:1 complex of Ca(4).calmodulin with IQ34 behaves as a compact globular species. The results demonstrate novel dynamic aspects of calmodulin-IQ interactions relating to the calcium regulation of motility of unconventional myosin. PMID- 12734702 TI - Raman study of the thermal behaviour and conformational stability of basic pancreatic trypsin inhibitor. AB - We have studied the thermal denaturation of native basic pancreatic trypsin inhibitor (BPTI) by monitoring the Raman bands in the 4000-400 cm(-1) range. In agreement with results obtained by calorimetry, a cooperative melting transition is observed starting at 75 degrees C. This transition is found to involve predominantly the unfolding of helical structures accompanied by beta aggregation, loss of hydrophobic interactions between side chains and changes in CSSC dihedral angles. However, salt bridge breaking starts near 40 degrees C, as deduced from the nu(s)(COO(-)) band and from the bands close to 1320 and 1345 cm( 1) which for the first time have been shown to be due largely to vibrations of the arginine guanidyl group in BPTI. The thermal stability is, hence, attributable to cooperative contributions from hydrophobic and backbone hydrogen bond interactions as well as from disulfide bonds. PMID- 12734701 TI - The aggregation behaviour of protein-coated particles: a light scattering study. AB - The different mechanisms involved in the aggregation of spherical latex particles coated with bovine serum albumin (BSA) have been studied using static and dynamic light scattering. These techniques assess the fractal dimension of the aggregates and their mean hydrodynamic radius. Particles with different degrees of surface coverage have been prepared. The net charge of the covered particles has been modified by varying the pH of the aqueous phase. The aggregation rate was measured and used to determine the importance of the different aggregation mechanisms that are responsible for these types of flocculation processes. At low and intermediate degrees of surface coverage, bridging flocculation is the principal aggregation mechanism irrespective of the electrical state of the protein-particle complexes. At high degree of surface coverage, however, weak flocculation is important only when the BSA molecules are at their isoelectric point. PMID- 12734704 TI - Occupational issues of allergic contact dermatitis. AB - Occupational contact dermatitis is often of multifactorial origin, and it is difficult to determine the relative significance of the various contributing factors. Contact allergies are relevant in 20-50% of recognised occupational contact dermatitis cases. The reported frequency in different studies varies, depending on differences in how occupational diseases are notified and recognised, in types of occupation in a geographical area, and the "quality" of the dermatological examination, including the accuracy of the diagnostic patch test investigation. However, the clinical relevance of the reported contact allergies is often uncertain. Many occupational contact dermatitis patients with documented contact allergies develop chronic eczema, in spite of work changes and attempted allergen avoidance. Recognition/non-recognition of a notified case may be based on circumstantial evidence, because of difficulties in the establishing of a firm proof of work exposure and subsequent development of skin disease. Reliable quantitative exposure measuring techniques are needed. Methods are developed for the measurement of exposure to allergens such as nickel and acrylates, which makes it possible for exposure-effect relationships to be established with increased certainty. For prevention of allergic contact dermatitis it was a major step forward, with mandatory ingredient labelling of cosmetic products. However, improved labelling of the presence of contact allergens in household and industrial products is needed. For the identification of hazardous contact allergenic compounds, guinea pig or mice assays are still required. The local lymph node assay (LLNA), which is an objective and sensitive mouse assay has now been internationally validated and accepted. PMID- 12734703 TI - Transitions from alkaline spots to regular bands during pH pattern formation at the plasmalemma of Chara cells. AB - A scanning pH-microprobe was used to study pH patterns near the surface of Chara corallina cells at various light intensities and during light-induced transitions from homogeneous pH distribution to alternating pH bands. In the irradiance (PAR) range 4-400 micromol quanta m(-2) s(-1), the sustained pH profiles consisted of alternating acid and alkaline bands with a characteristic length of 7-10 mm and pH shifts as large as 2-3 units. At lower irradiance, the number of alkaline bands decreased while the amplitude of remaining peaks stayed high. On cyclic changes in light intensity, a hysteresis of pH banding was observed: the pH bands tolerated low irradiance in weakening light, but higher irradiance was required for their emergence after dark adaptation of the cell. The pH profiles measured for different paths of electrode scanning suggest that the pH pattern at low light level represents patches coexisting with bands. The exposure of the cell to high-intensity light led to formation of radially symmetrical bands. Transformations of the pH pattern induced by lowering the light intensity were similar to those induced by transcellular electric current (1.5-3 microA). The data suggest that band formation at the plasmalemma of Chara cells proceeds through the initial appearance of multiple patches with a localized H(+) transporting activity and subsequent spot rearrangements (fusion, deletions, widening), leading to establishment of alternating bands. PMID- 12734705 TI - Multifocal ERG with the scanning laser ophthalmoscope: query on the ideal configuration for attaining high resolution and result stability. PMID- 12734706 TI - Recovery from hepatic retinopathy after liver transplantation. AB - BACKGROUND: In chronic liver disease the neuroglial cells may be affected by neurotoxic metabolites which, in turn, could be expected to affect neuronal functions. The aim of this study was to evaluate the electroretinograms (ERG) from patients before and after liver transplantation, in order to study possible functional changes of the retina. METHODS: Twelve patients with liver cirrhosis underwent routine ophthalmological examination and ERG before and after successful liver transplantation. Laboratory parameters, including ammonia, aspartate aminotransferase (AST), bilirubin, and cholinesterase, were compared. Patients were grouped according to the Child classification: three patients were Child A, six were Child B, three were Child C. RESULTS: Most obvious ERG abnormalities were found in patients with cirrhosis Child C. Before transplantation 7 of 21 ERG parameters were out of the normal range, but in the follow-up examination after transplantation only one parameter was not within the normal range. Significant ( P<0.05) postoperative improvements were found for the latencies of scotopic, mesopic and photopic b-waves and mesopic a-waves and for the photopic implicit time. Patients in the Child B group revealed less changes in the ERG. Before the transplantation only one parameter of the ERG was out of the normal range. All postoperative parameters were within the normal range. At 40+/-9 months after the liver transplantation a significant decrease in serum ammonia levels, AST and bilirubin and a significant increase in cholinesterase levels were observed. CONCLUSION: Patients with restored liver function after liver transplantation showed significantly improved ERG parameters. Our data suggest a recovery of the cells involved in hepatic retinopathy, including the Muller (glial) cells. PMID- 12734707 TI - Concentric retinal pigment epithelium atrophy after a single photodynamic therapy. AB - PURPOSE: To report a case with concentric retinal pigment epithelium (RPE) atrophy after a single photodynamic therapy (PDT). METHODS: We report a case of a 33-year-old female patient who developed RPE atrophy after a single standard PDT for treatment of a juxtafoveal, predominantly classic choroidal neovascularization (CNV). RESULTS: After a single PDT treatment, visual acuity increased from 20/50 to 20/20. Six weeks after PDT, a concentric area of RPE atrophy was clearly visible on fluorescein angiogram. This circular area corresponded to the 3500 microm diameter of the laser spot used in the PDT treatment. The visual acuity and the RPE atrophy remained stable over the follow up period of 3 years. CONCLUSIONS: We are unable to explain the exact mechanism of the observed RPE changes; however, they did not lead to loss of visual acuity. Different reasons for the RPE atrophy such as collateral damage of the choriocapillaris with a subsequent secondary RPE atrophy, a direct photochemical effect due to the early localization of the photosensitizer in the RPE, or a depigmentation or photobleaching of the RPE, which led to a window defect in the fluorescein angiogram without loss of the major functional properties of the RPE, are possible mechanisms involved in the development of the documented lesion. PMID- 12734708 TI - Spatial visual filtering in diabetes mellitus. AB - PURPOSE: To investigate spatial visual filtering in a group of diabetic patients and compare the results with those of a group of controls. METHODS: The luminance threshold of a moving 2 degrees achromatic target, viewed against a 17 degrees achromatic background grating, was measured as a function of grating periodicity from 0.21 to 31.4 cpd in 22 diabetic patients and 12 controls, giving a response characteristic of the spatial function of a sustained-response type of visual channel. A previously published model of spatiotemporal filtering, integrating photoreceptor kinetics with difference-of-Gaussian circularly symmetric receptive fields, was used to analyse the data. METHODS: The model gave a good fit to the data in the control group, with a mean central space constant of 0.046 degrees and centre:surround ratio of 1:5.2 and mean R(2)=0.78 (SD 0.12). The mean central space constant in the diabetic group was 0.051 degrees and the centre:surround ratio 1:4.2, although best fit was significantly worse, at R(2)=0.54 (SD 0.19), P=0.001. The best fit for diabetic subjects with grade 2 maculopathy was significantly worse than for those with no maculopathy ( P=0.03). CONCLUSION: The study demonstrates a disruption of circularly symmetric centre-surround receptive field structure of the sustained-response channel in the diabetic retina to a degree that is consistent with the retinal level of anatomical change in diabetic maculopathy. PMID- 12734710 TI - Reconstruction of acute anterior cruciate ligament injuries: a prospective, randomised study of 40 patients with 7-year follow-up. No advantage of synthetic augmentation compared to a traditional patellar tendon graft. AB - BACKGROUND: In a prospective, randomised study, we compared the results of anterior cruciate ligament reconstruction using the conventional medial patellar tendon strip procedure with the Kennedy ligament augmentation device (LAD) over the-top augmentation technique. The aim of the study was to see if the addition of this device, with its specific potential complications (infection, synovitis, mechanical failure) and increased costs, gave better functional stability and less donor site morbidity. METHODS: Forty patients ( n=20+20) with high physical demands, anterior cruciate ligament rupture less than 3 weeks old, and positive Lachman and pivot shift tests were included in the study. They were randomised by instructions in a sealed envelope that was randomly chosen. Postoperatively, they were immobilised in a plaster of Paris cast for 2 weeks. Full weight-bearing was not allowed until 6 weeks after the operation. Strength training on the operated side started when movement was restored to nearly normal, usually after 8-10 weeks. Return to full sport activity was allowed 1 year after the operation. Clinical and arthrometric follow-up was performed yearly using the Lysholm functional score, Tegner activity score, KT-1000 testing, Lachman and pivot shift tests. Follow-up lasted 3-9 years (mean 7 years). RESULTS AND CONCLUSIONS: At last follow-up we found no statistical difference in stability tests, functional or activity scores; both groups showed stable improvement concerning scores and arthrometry compared with the preoperative values. No advantages were associated with the use of the Kennedy LAD, and we do not recommend it or a similar device for uncomplicated cases. New augmentation devices are simply launched onto the market, and we recommend caution and thorough evaluation in prospective, randomised studies before they are adopted into use. PMID- 12734711 TI - Evaluation of the transverse metatarsal arch of the foot with gait analysis. AB - BACKGROUND: The existence of the transverse metatarsal arch (TMA) of the foot is a point of controversy. According to Kapandji, TMA of the foot elevates the 2(nd) to 4(th) metatarsal heads. Some authors suggest the existence of TMA, while others suggest that there is no functional metatarsal arch of the foot. In this study, we evaluated the existence of TMA of the foot and weight distribution on the metatarsal heads with the EMED-SF (Novel H, Munich, Germany) plantar pressure analysis system. METHODS: The test was performed with 16 volunteers. According to the three functional columns of the foot, the metatarsal region of pressure picture obtained from the EMED-SF system was divided into three regions called 'masks'. Mean pressures in the masks were calculated at the mid-stance phase. RESULTS: The highest mean pressure recorded was located at the 2nd to 3rd metatarsal heads (7.96 N/cm(2)), and the second highest pressure was at the heel (6.55 N/cm(2)). The pressures of the 1st metatarsal and 4th-5th metatarsal heads were 4.86 and 6.26 N/cm(2), respectively. The difference between the pressure distributions under metatarsal heads was statistically significant ( p=0.000). CONCLUSION: According to our results, TMA of the foot does not exist as described by Kapandji. PMID- 12734709 TI - Sequence polymorphisms within the human mitochondrial genes MTATP6, MTATP8 and MTND4. AB - By sequencing the control region of mitochondrial DNA, the majority of human DNA samples can be differentiated. A further increase in differentiation probability may be possible, e.g. by extending the sequenced region to coding regions of the mitochondrial genome. Restriction to those positions that do not result in a change of the amino acids guarantees that the information thus obtained does not refer to phenotypically relevant information. In the present study the sequence data of the mitochondrial genes MTATP6, MTATP8 and MTND4 were collected from 109 subjects and analyzed in order to define variable positions suitable for identification purposes. There were 32 variable base positions among 850 bases studied from MTATPase genes and 1,200 bases of the MTND4 gene showed 28 variable positions. "Hot spots" for base exchanges were found in both regions and one position (position 11,719 in the MTND4 gene) seems to be suitable for SNP investigation for forensic purposes. PMID- 12734712 TI - The angle velocity reproduction test (AVRT) as sensorimotor function of the glenohumeral complex. AB - BACKGROUND: The purpose of the present study was to develop and evaluate the angle velocity reproduction test (AVRT) in the glenohumeral joint. METHODS: In 46 volunteers with healthy shoulders, a test was devised in order to record the angle velocity perception in the shoulder. One arm was moved passively by means of a Cybex 6000 unit. The subjects were asked to perform the same movement as precisely as possible with the contralateral arm without visual control. The movement was recorded with a contact-free motion analysis system with a digital infrared camera and evaluated by aid of a movement analysis program. Starting at 20 degrees, the movement was an anteversion (anterior flexion) of 110 degrees. The sector between 20 degrees and 90 degrees was analyzed in detail. Different parameters were documented which represented different aspects of the sensorimotor function. The predetermined angle velocity of the Cybex setting on the contralateral arm amounted to exactly 53 degrees /s. In order to observe validity and reliability, 10 additional measurements with visual control as well as 21 double measurements were performed. RESULTS: The comparison of the measurements with and without visual control yielded highly significantly better results for those courses with visual control with regard to all those parameters which represent the perception of velocity. The values which describe the evenness of the movements were weak and/or insignificant. CONCLUSION: The introduced AVRT appears suitable to test dynamic sensorimotor abilities. PMID- 12734713 TI - Histological findings of tendon-bone healing following anterior cruciate ligament reconstruction with hamstring grafts. AB - BACKGROUND: The purpose of the study was the histological examination of tendon bone healing of hamstring grafts after anterior cruciate ligament (ACL) reconstruction. METHODS: During five arthroscopies done 6-14 months after ACL reconstructions, biopsies of the wall of the former drilled femoral canal were obtained. Four patients were primarily operated on using a suspending device (Endobutton, Acufex Microsurgical, Mansfield, MA, USA, and Transfix, Arthrex, Naples, FL, USA) for femoral fixation, one patient was reconstructed with a biodegradable interference screw directly inserted between the tendon and the wall of the canal. Biopsies were obtained using a tube harvester during re arthroscopy. Three grafts were stable, two grafts were unstable, and revision of the ACL was performed. RESULTS: Histologically, in the four cases of reconstruction with a button or a rectangular pin, biopsies resembled granulation tissue without insertion of fibers between the tendon tissue and the bony wall. A wide area of woven bone was noted adjacent to the pre-existing lamellar bone. In contrast, the tendon-bone junction in the knee reconstructed with a biodegradable interference screw resembled a zone of metaplastic fibrous cartilage between the tendon graft and the lamellar bone. Collagen fibers connecting the tendon-bone interface occurred under polarized light microscopy. CONCLUSION: We conclude that the use of hamstring grafts for ACL reconstruction can lead to different histological pattern of tendon-bone healing. Micromotion of the hamstring graft inside the drilled canal can be play a role in tendon-bone healing. PMID- 12734714 TI - Why patients choose regional anesthesia for orthopedic and trauma surgery. AB - BACKGROUND: While both surgeons' and anesthesiologists' preference of regional over general anesthesia is increasing, the patients' preference remains limited. Little is known about why patients choose regional anesthesia. The aim of our study was to answer this question with regard to orthopedic and trauma surgery. METHODS: The study was carried out prospectively from 1999 to 2001 and included 238 patients scheduled for arthroscopy of the lower limb or other orthopedic or trauma surgery. All patients were informed about regional and general anesthesia in a pre-anesthesia interview and subsequently chose the method they preferred. RESULTS: Curiosity was the main reason why patients chose regional anesthesia for arthroscopy ( n=155, p<0.0001). For all other types of orthopedic or trauma surgery ( n=83), patients chose regional anesthesia to avoid postoperative pain ( p<0.01) and/or the side-effects of general anesthesia ( p<0.0001). Younger ( n=128) and low-risk ( n=184) patients chose regional anesthesia because they were curious ( p<0.01 and p<0.001, respectively), while older ( n=110) and unhealthier patients ( n=56) did so for safety ( p<0.01 and p<0.001, respectively). CONCLUSIONS: Patients choose different types of regional anesthesia for different reasons. While spinal anesthesia and femoral and sciatic block were chosen for curiosity reasons, brachial plexus block was chosen to avoid the side effects of general anesthesia. The patients' choice of regional anesthesia for orthopedic and trauma surgery is significantly influenced by the type of surgery, age, and health. PMID- 12734715 TI - The angiogenic peptide vascular endothelial growth factor (VEGF) is expressed during the remodeling of free tendon grafts in sheep. AB - BACKGROUND: Angiogenesis is important for the remodeling of autologous tendon grafts. A sheep model was used to examine the expression of the angiogenic peptide vascular endothelial growth factor (VEGF) in autologous tendon grafts after anterior cruciate ligament (ACL) reconstruction. METHODS: Merino sheep underwent ACL reconstruction with an autologous Achilles tendon split graft. VEGF and its receptors, the Fms-like tyrosine kinase receptor (FLT-1, VEGFR-1) and the kinase insert domain-containing receptor KDR (VEGFR-2/FLK-1) were detected immunohistochemically. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect which of the different VEGF splice forms are expressed during ACL remodeling. RESULTS: At 6 weeks, light microscopy showed zones with hypocellular necrotic graft tissue in the central part surrounded by hypercellular and hypervascularized reparative tissue invading the former graft tissue from the periphery. In contrast to the necrotic tissue, all cell types of the reparative tissue labeled strongly positive for VEGF. The VEGF receptors FLT 1 and KDR could be detected on endothelial cells of blood vessels. At 12 weeks, the complete graft diameter showed an increased vascular density (anti-factor VIII immunoreactivity), but zones of non-remodeled former graft tissue could not be found. Fusiform fibroblasts labeled strongly for VEGF. At 24 weeks, VEGF immunostaining decreased, and at 52 and 104 weeks, the grafts were largely VEGF negative. RT-PCR supported the immunohistochemical results regarding VEGF expression and showed further that the splice variants VEGF(120 )and VEGF(164) are expressed during angiogenesis during the remodeling of tendons. CONCLUSION: We conclude that the angiogenic peptide VEGF plays a role in tendon graft remodeling. PMID- 12734716 TI - Anatomical reconstruction of chronic lateral ligament injury of the ankle using pedicle tendon of the extensor digitorum longus. AB - BACKGROUND: Since 1989, we have performed ligament reconstruction using the extensor digitorum longus tendon as a pedicle graft for patients with chronic lateral instability of the ankle. METHODS: Of those patients who underwent arthroscopy during the operation, thirteen joints were evaluated at an average follow-up of 7 years and 2 months. The overall average postoperative score was 95.4 points on Karlsson's score for the ankle ligament. Instability scores improved by 8.5-24.6 points. Our anatomical reconstructive surgery showed generally excellent functional results, with cosmetic advantages because of a short incision and preservation of the entire peroneus brevis tendon, which plays an important role in extra-articular dynamic stabilization. RESULTS: At final follow-up, pain was present in one joint in the group without chondral injury at the time of reconstruction ( n=6). However, pain was still present at the final follow-up in five of the group with chondral injury ( n=7). Chondral injury incurred during ligament reconstruction may have caused the residual ankle pain even after 7 years of postoperative follow-up. There was a statistically significant relationship between chondral injury and ankle pain at final follow up. PMID- 12734717 TI - Primary ACL reconstruction with fresh-frozen patellar versus Achilles tendon allografts. AB - BACKGROUND: In recent years, there has been an interest in the use of allografts as an alternative graft for anterior cruciate ligament (ACL) reconstruction to reduce potential donor-site morbidity resulting from the harvest of autogenous tissue. Nevertheless, in the literature, the use of allografts for primary ACL reconstruction is controversial due to a higher failure rate and the potential risk of disease transmission. METHOD: In this retrospective study, we evaluated the clinical outcome of 251 fresh-frozen patellar vs Achilles tendon allografts for primary ACL reconstruction. Patients (average age 39 years) were operated on between 1993 and 1998, and the mean follow-up was 37.7 months (range 24-74 months). We were able to follow up 225 patients (89.6%). According to the different types of allograft, we divided the patients into two groups: group P with patellar bone-tendon-bone allograft (BTB; n=183) and group A with Achilles bone-tendon allograft ( n=42). Clinical evaluation consisted of a history, an examination, IKDC Score, Cincinnati Knee Score (CKS), Cincinnati Sports Activity Scale (CSAS), KT-1000 testing, and standardized X-rays. RESULTS: According to the IKDC, the outcome was normal or nearly normal in 75.3% in group P and 76.2% in group A. Overall rating according to the CKS was an average of 85 in group P and 82.9 in group A. CSAS was 79.6 in group P and 84.8 in group A. The objective stability measured with the KT-1000 showed an average side-to-side difference of 2.1 mm in group P and 2.0 mm in group A. 4.4% of group P and 2.5% of group A were considered a laxity failure, and 10.4% of group P and 4.8% of group A re-ruptured the reconstructed ACL. In summary, there was a significantly higher failure rate ( p<0.001) in group P compared with group A. CONCLUSION: Satisfactory clinical results can be achieved with the use of allografts for primary ACL reconstruction. Comparing Achilles tendon and patellar BTB allografts, the Achilles tendon-bone allograft seems to be advantageous for ACL reconstruction as the failure rate was significantly lower. Nevertheless, the total failure rate appears to be much higher compared with autogenous ACL reconstruction, indicating that the use of an allograft for routine uncomplicated primary ACL reconstruction offers few advantages. Therefore, autograft tissue remains our graft of first choice for this procedure. We advise reserving allografts for revision procedures where suitable autogenous tissues have been previously compromised, where a contraindication for autogenous tissue harvest exists, or for multiple ligament surgery. PMID- 12734718 TI - Epidemiology of posterior cruciate ligament injuries. AB - BACKGROUND: The epidemiology of posterior cruciate ligament (PCL) injuries has not been well clarified. Isolated and combined PCL injuries are a frequently missed diagnosis. A better understanding of typical injury mechanisms may help in more accurate diagnosis of these injuries. METHODS: In this study the epidemiology of PCL insufficiency in 494 patients was retrospectively analysed. Stress-radiography was used to quantify posterior tibial displacement. RESULTS: The mean age at the time of injury was 27.5+/-9.9 years. Traffic accidents (45%) and athletic injuries (40%) were the most common injury causes. Motorcycle accidents (28%) and soccer-related injuries (25%) accounted for the main specific injury causes. The most common injury mechanisms were dashboard injuries (35%) and falls on the flexed knee with the foot in plantar flexion (24%). The mean side-to-side difference of posterior tibial displacement on posterior stress radiographs in 90 degrees of flexion was 13.4+/-4.7 mm. According to the posterior displacement values, 232 (47%) patients had isolated PCL ruptures, while 262 (53%) patients with a posterior displacement of >12 mm were classified as having a combined posterior instability. There were significantly more combined PCL lesions due to vehicular trauma as compared with athletic trauma ( p<0.0001). CONCLUSIONS: In many PCL lesions, initiation of an adequate treatment regimen is delayed despite typical injury mechanisms and symptoms. In the future, a better understanding of the epidemiology of PCL injuries should enable us to diagnose the injury more reliably through a detailed history and a thorough physical and radiographic examination in the acute setting. PMID- 12734719 TI - Improved production of ( S)-ketoprofen ester hydrolase by a mutant of Trichosporon brassicae CGMCC 0574. AB - An efficient screening method following UV mutagenesis yielded a high frequency of improved mutants of Trichosporon brassicae CGMCC 0574, a wild-type esterase producer capable of enantioselectively hydrolyzing the ethyl ester of ketoprofen [2-(3-benzoylphenyl) propionic acid]. The mutant had an activity 1.8-fold higher than the wild type and was stable in its enzyme production for ten serial transfers. As the best single carbon source, isopropanol improved the specific activity of the enzyme 5-fold; and this did not result from the effect of cell permeabilization. An 18-h culture grown on a medium containing 0.5% glucose plus 0.5% isopropanol produced 3-fold as much esterase as a culture grown on 1% glucose. PMID- 12734720 TI - A method for separation and determination of cytokinin nucleotides from plant tissues. AB - Recent progress in the study of cytokinin metabolism in plants indicates that quantitative analysis of cytokinin nucleotides is essential for elucidation of early steps of the biosynthetic pathway. However, traditional procedures for purification and quantification of cytokinin cannot discriminate the various nucleotides. We describe here a method for separation and determination of cytokinin nucleotides through a series of anion-exchange column chromatography steps followed by liquid chromatography-mass spectrometry. This method enabled us to analyze the amount of each species of cytokinin nucleotide in plant tissues. PMID- 12734722 TI - Detection of several Mycoplasma species at various anatomical sites of homosexual men. AB - In order to determine the colonisation patterns of several Mycoplasma species in homosexual men, urethral, oral and rectal specimens from 10 homosexual men with acute non-gonococcal urethritis (NGU) and 18 without NGU were examined using sensitive methods. Mycoplasma hominis and Ureaplasma urealyticum existed in both groups, which is in keeping with previous studies of heterosexual men. Mycoplasma genitalium was detected in the rectum of both NGU-positive and NGU-negative men and in the urethra of one man with chlamydia-negative NGU, but not in those without urethritis. Mycoplasma fermentans was found in the throat and rectum only and Mycoplasma penetrans in all three anatomical sites. In contrast, Mycoplasma pirum was found in the rectum only, that is, in 5 of the 28 men studied. Infrequent examination of this site is a possible explanation for previous failures to detect Mycoplasma pirum at a mucosal surface. PMID- 12734723 TI - Lack of portosystemic bacterial translocation in patients with liver cirrhosis after placement of transjugular shunt. AB - The purpose of this study was to clarify whether bacteria are transferred from the portal venous system into central venous blood during the placement of a transjugular portosystemic stent shunt (TIPSS). TIPSS was created in 30 consecutive cirrhotic patients for recurrent variceal bleeding ( n=12), refractory ascites ( n=16), or hepatorenal syndrome ( n=2). Microbiological analysis was performed prospectively on central venous blood before and on portal venous blood immediately after puncture of the portal vein. Twenty minutes after the placement of TIPSS, another sample of central venous blood was obtained. None of the first two sets of blood cultures showed bacterial growth, so that no bacterial transfer was seen at the time of TIPSS placement. Four of the third sets of blood samples showed skin and mouth flora, interpreted as iatrogenic contamination. PMID- 12734724 TI - Novel mutations of neurofibromatosis type 1 gene in small cell lung cancers. AB - PURPOSE: Mutations in the GTPase-activating protein-related domain (GRD) of the von Recklinghausen neurofibromatosis type 1 (NF1) gene have been reported in several tumors that were not previously associated with NF1. We analyzed 37 cases of lung cancer, including 9 cases of small cell carcinoma, to detect mutations in this domain. METHODS: DNA was extracted from the tumors, and single-strand conformation polymorphism and direct sequencing were used to detect mutations. RESULTS: Three cases of small cell carcinoma had mutations in NF1-GRD. There were two incidences of an A --> G transition at the second base of codon 1415 resulting in Glu --> Gly substitution, and one incidence of an A --> G transition at the third base of codon 1411 (Pro) without amino acid substitution. All three patients had both hilar and mediastinal lymphatic metastasis. CONCLUSIONS: A mutation in this region could imply a poor prognosis. PMID- 12734721 TI - Management of Aspergillus osteomyelitis: report of failure of liposomal amphotericin B and response to voriconazole in an immunocompetent host and literature review. AB - Presented here is a case of Aspergillus osteomyelitis in an immunocompetent patient that progressed despite surgery and prolonged treatment with liposomal amphotericin B; the report is followed by a review of the literature. The review of this case and 41 similar cases found an overall cure rate of 69%. The importance of surgery when amphotericin B is used as first-line therapy is indicated by a 14% cure rate when amphotericin B is used alone compared to 75% when combined with surgery. When therapy is failing or surgery is contraindicated, dose escalation using a lipid formulation was not effective. On review, the addition of another agent, in particular 5-fluorocytosine, appears to be more beneficial. The patient reported here responded rapidly to voriconazole, a promising new antifungal agent for Aspergillus infections. PMID- 12734725 TI - Frequency of left atrial myxoma with concomitant coronary artery disease. AB - PURPOSE: Simultaneous coronary artery bypass grafting with a resection of left atrial myxoma has been rarely reported. The ages and the symptoms of patients who have left atrial myxomas and coronary artery disease are similar. In this report, we present our cases of left atrial myxoma with concomitant coronary artery disease who were all treated surgically. METHODS: Between September 1998 and January 2001, 11 patients were surgically treated after being diagnosed to have left atrial myxoma. Routine coronary angiography was performed on all patients preoperatively. In four patients concomitant coronary artery disease was identified. At surgery we performed coronary artery bypass grafting after a resection of left atrial myxoma in three patients. RESULTS: All patients were weaned from cardiopulmonary bypass without any difficulty. The postoperative course was uneventful. The follow-up period was 17 +/- 10 months (range 3-32 months). All patients were symptom-free and no recurrence of myxoma was detected. CONCLUSION: Based on our experience, cardiovascular surgeons should be aware of the concomitance of these diseases. It is therefore recommended that coronary angiography should be performed on all patients who present with left atrial myxomas. PMID- 12734726 TI - Management of inflammatory tracheoesophageal adhesions during transhiatal esophagectomy. AB - PURPOSE: Mediastinal adhesions, caused by the transerosal spread of disease, inflammatory diseases, or preoperative chemoradiotherapy, can result in diffi culties and major complications during transhiatal esophagectomy (THE). However, few studies have specifically addressed the incidence and management of inflammatory adhesions encountered during THE. METHODS: We retrospectively analyzed the operative details and postoperative outcome of 70 patients who underwent THE between 1998 and 2000. Patients with inflammatory tracheoesophageal adhesions were identified and their operative records were reviewed for operative findings, intraoperative management, morbidity, and mortality. Patients with upper thoracic esophageal tumors and direct tumor spread to the airways were excluded from the analysis. RESULTS: Eight (11.4%) of the 70 patients had inflammatory adhesions and esophagectomy was possible in 7 of these 8 patients. Due to poor pulmonary status, one patient required conversion to the trnsthoracic approach, but the other six were managed transhiatally. Three patients underwent subtotal esophagectomy, the esophagectomy was completed by the inversion extraction technique in two, one required extended transhiatal dissection, and esophagectomy could not be completed in one due to dense inseparable adhesions between the trachea and esophagus. There was no major airway injury or bleeding. CONCLUSIONS: Inflammatory tracheoesophageal adhesions may be encountered in patients undergoing THE in developing countries. Thus, an awareness of the possibility of these adhesions and suitable modifications of the operating procedures are necessary to prevent major complications. PMID- 12734727 TI - Impact of a clinical pathway and standardization of treatment for acute appendicitis. AB - PURPOSE: Acute appendicitis is one of the most common surgical diseases. Simple and precise guidelines for treating acute appendicitis are necessary for improving the treatment outcome of this disease. The purpose of this study was to determine the impact of a clinical pathway and standardization of treatment for acute appendicitis at our hospital. METHODS: The clinical pathway and standardization of treatment for acute appendicitis were introduced to our hospital in January 2000. We compared the length of hospitalization, postoperative stay, hospital costs, and operation time during the years before and the years after their introduction. RESULTS: There was no significant difference in the clinical characteristics of the 73 patients in the control group and the 112 patients in the pathway group. There were 6 (8.2%) and 24 (21.4%) cases of perforated appendicitis in the respective groups. The mean length of hospitalization ( P < 0.001), postoperative stay ( P < 0.001), and hospital costs ( P < 0.01) were significantly less in the patients in the pathway group who underwent surgery. CONCLUSION: Our clinical pathway and standardization of treatment for acute appendicitis proved effective for treating patients with acute appendicitis and minimizing costs without compromising patient care. PMID- 12734728 TI - Functional results after endoscopic subtotal cortical-sparing adrenalectomy. AB - PURPOSE: We examined the required amount of residual adrenal tissue and whether an intact adrenal vein are necessary to achieve sufficient function after endoscopic subtotal adrenalectomy. METHOD: Endoscopic subtotal adrenalectomy was performed in 14 patients. Two patients underwent unilateral subtotal and contralateral total adrenalectomy and another two patients underwent unilateral subtotal adrenalectomy after contralateral total adrenalectomy several years earlier. We analyzed the postoperative serum levels of cortisol and adrenocorticotropic hormone (ACTH). Patients with bilateral tumors underwent an ACTH test. RESULTS: We had to cut the main adrenal vein in ten patients, and less than one third of the adrenal gland was left in situ in four patients. Subtotal adrenalectomy was performed unilaterally in two patients with bilateral tumors. One third of the adrenal gland was preserved in these patients, and also in the two patients with unilateral subtotal adrenalectomy after previous contralateral total adrenalectomy. The postoperative ACTH test confirmed satisfactory adrenocortical function. During the follow-up period of about 24 months no recurrent tumors have been found. CONCLUSION: Subtotal cortical-sparing adrenalectomy can be successfully performed laparoscopically. The venous drainage of the main adrenal vein does not seem to be crucial for sufficient adrenocortical function. We estimate that leaving about one third of the entire adrenal gland as remnant adrenal tissue will result in sufficient function. PMID- 12734729 TI - Experimental and clinical studies investigating the efficacy of distal anastomosis with patch plasty in bypass operations with expanded polytetrafluoroethylene grafts. AB - PURPOSE: We investigated the efficiency of distal anastomosis with patch plasty (DAPP), both experimentally and clinically. METHODS: In our experimental study, dogs were divided into two groups: a control group in which anastomosis was performed without DAPP ( n = 7), and a DAPP group in which DAPP was performed at the distal anastomosis ( n = 7). In our clinical study, 169 femoropopliteal bypasses were divided into three groups and analyzed. In one group, the saphenous vein was used (SVG group, n = 65); in one group, an expanded polytetrafluoroethylene (ePTFE) graft was used without DAPP (ePTFE group, n = 64); and in one group, an ePTFE graft was used with DAPP (DAPP group, n = 40). RESULTS: In the experimental study, the ratio between the area of thrombus adherence and the entire area of the intraluminal surface of the graft, defined as the thrombus covering ratio, was 48.9% in the control group and 30.2% in the DAPP group. The ratio in the DAPP group was significantly lower than that in the control group. In the clinical study, although there were no significant differences among the three groups in cumulative patency rates of the femoral above-knee popliteal arterial bypasses, the patency in the DAPP group was excellent. The cumulative patency rates of the femoral below-knee popliteal arterial bypasses in the ePTFE group were significantly lower than those in the other two groups. CONCLUSION: There results suggest that the addition of DAPP may achieve excellent graft patency. PMID- 12734730 TI - Spontaneous retropharyngeal hematoma of a parathyroid cyst: report of a case. AB - A 41-year-old woman presented with severe and sudden anterior neck swelling, pain, and dysphagia. Computed tomography (CT) scan and ultrasound of the neck showed a giant mass in the retropharyngeal space, displacing the trachea and esophagus anteriorly. Aspiration cytology was done, following which extensive cervical and chest ecchymosis occurred and her symptoms immediately improved. A repeat CT scan demonstrated that the cervical giant mass had vanished, but there was a residual mass in the left paratracheal space. Exploratory surgery of the neck revealed a parathyroid cyst with severe adhesion to the surrounding tissues. We considered that a ruptured parathyroid cyst had induced massive hemorrhage into the cervical tissues and mediastinum, but that the hemorrhage had been absorbed. Extracapsular hemorrhage from a parathyroid adenoma or cyst is rare, especially from a parathyroid cyst. In fact, to our knowledge, this represents only the third case of symptomatic spontaneous bleeding of a parathyroid cyst. Nevertheless, this entity should still be considered in the differential diagnosis of all rapidly progressing retropharyngeal masses. PMID- 12734731 TI - Characterization of apoptosis-related molecular changes in a desmoid tumor of the chest wall: report of a case. AB - Desmoid tumors are uncommon benign tumors composed of fibrous tissue, which originate from aponeuroses. Little is known about their molecular pathogenesis and the reason that they recur. We report the case of a 20-year-old man with a recurrent desmoid tumor of the chest wall, focusing on our analysis of the apoptosis and its related molecular events. Immunohistochemical examination showed higher expression of antiapoptotic Bcl-2, Bcl-XL, survivin, and the transcription factor, NF-kappaB, in the recurrent tumor than in the adjoining normal tissue. Proapoptotic Bax was not detected in the tumor. Similar findings were obtained in the original primary tumor. Both tumors had a low apoptotic index according to the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay. These changes occurred in the absence of cell proliferation, shown by the absence of both Ki-67 staining and increased telomerase activity. This derangement of apoptosis gives the aggressive desmoid tumor cells a proliferative advantage, and presumably, forms the basis of its high recurrence rate. Therefore, inhibitors of apoptosis proteins (IAPs) may be useful for predicting recurrence. The regulation of apoptosis by antisense therapy against these inhibitors could prove beneficial for overcoming repeated recurrence, even after surgery. PMID- 12734732 TI - Simultaneous open abdominal aortic replacement and thoracic stent-graft placement: report of a case. AB - A 77-year-old man with severe chronic obstructive pulmonary disease was admitted to our hospital for surgical treatment of a proximal descending thoracic aortic aneurysm (dTAA) and an infrarenal abdominal aortic aneurysm (AAA). The patient had poor respiratory function; however, a simultaneous abdominal aortic replacement and thoracic stent-graft placement were successfully performed without any complications. This case report demonstrates that simultaneous abdominal aortic replacement and thoracic stent-graft placement for multiple aneurysms may be feasible and can safely be performed in selected high-risk patients, despite the many problems associated with the treatment of aortic aneurysms using stent grafts. PMID- 12734733 TI - Infected aneurysm of the ascending aorta in childhood: report of a case. AB - Infected aneurysms of the thoracic aorta are rare in children, and many are not diagnosed until autopsy. We report the case of a 3-year-old girl in whom an infected aneurysm of the ascending aorta was successfully repaired by Dacron patch angioplasty under cardiopulmonary bypass. PMID- 12734734 TI - Totally laparoscopic distal gastrectomy using the hand-sewn Billroth-I anastomotic technique: report of a case. AB - The development of more sophisticated instruments has enabled advanced laparoscopic surgery. We recently devised a totally laparoscopic method of performing Billroth-I hand-sewn anastomosis and established this technique in an animal training model. This report presents the case of a 50-year-old man in whom totally laparoscopic distal gastrectomy was successfully performed for gastric cancer, using the hand-sewn Billroth-I anastomotic technique. The patient was admitted with gastric cancer in the angle of the stomach and underwent laparoscopic distal gastrectomy with radical lymph node dissection. After the resected specimen was extracted through the small incision, a Billroth-I anastomosis was performed laparoscopically by the hand-sewn technique using the Albert-Leinbert method. The patient was discharged on the seventh postoperative day without any intra- or postoperative complications. Laparoscopic hand-sewn anastomosis was performed safely and allowed for quick recovery and good cosmesis in this patient. PMID- 12734735 TI - Adenocarcinoid of the appendix: report of two cases. AB - Adenocarcinoid of the appendix is a rare tumor with the histological features of both adenocarcinoma and carcinoid tumor. However, its biological behavior and malignant potential are still unclear. We treated two patients with this unusual tumor; a 60-year-old man and a 79-year-old woman. Both patients were initially diagnosed with acute appendicitis followed by an appendectomy. At surgery, the appendix was seen to be acutely inflamed without any macroscopic signs of tumor. Postoperative histological analysis revealed an adenocarcinoid tumor in the appendix, which had spread diffusely into its wall without forming a mass. Immunohistochemical staining with p53, MIB-1, bcl-2, and carcinoembryonic antigen suggested that neither of these tumors were particularly aggressive. Adenocarcinoid of the appendix is a rare tumor, which is very difficult to diagnose preoperatively and even macroscopically, making histological examination essential. PMID- 12734736 TI - Port site metastasis of sigmoid colon cancer after a laparoscopic sigmoidectomy: report of a case. AB - A 75-year-old woman with TNM stage I (T2, N0, M0) sigmoid colon adenocarcinoma underwent a laparoscopy-assisted sigmoidectomy. Five months later, the patient developed two subcutaneous metastases at two trocar sites. A second operation was performed and two abdominal wall masses were resected, but the operation revealed no evidence of peritoneal dissemination, liver metastasis, or lymph node metastasis. The histological findings confirmed the diagnosis of metastatic carcinoma of the sigmoid colon. Large-scale studies have shown the actual rate of port site metastasis to be similar to that observed in open surgery, and therefore the rate is much lower than that initially reported in small studies. However, the metastatic mechanism has not yet been clarified. These results suggest that port site metastasis can be prevented as effectively as during open surgery by utilizing accurate operating skills based on the sound principles of tumor surgery. Therefore, it is very important that appropriate operating procedures and an effective training system are established. PMID- 12734737 TI - Sigmoid colon perforation as an unusual complication of Behcet's syndrome: report of a case. AB - A 47-year-old man with long-standing Behcet's syndrome presented with an acute abdomen, and was found to have perforation of the sigmoid colon. Laparotomy revealed gangrenous changes in the sigmoid colon and perforation in the center of the affected segment. This is a very rare complication of Behcet's disease, and we report this case to stress the importance of performing careful abdominal examination while evaluating patients with Behcet's disease. PMID- 12734739 TI - Early-onset liver abscess after blunt liver trauma: report of a case. AB - A 23-year-old male patient underwent nonoperative management for his blunt liver trauma as he was hemodynamically stable without any signs of peritonitis initially after injury. A fever of 39.5 degrees C and severe right upper quadrant abdominal pain developed on the second day, and an abdominal computed tomography (CT) scan showed the formation of a gas-containing liver abscess in the traumatized liver. An emergency laparotomy revealed a foul-smelling liver abscess at the traumatized site, which was finally disclosed to be the result of a Clostridium species infection. A liver abscess is a rare complication following the nonoperative management of liver injury, and such an occurrence is even more rare within 1 day after injury. A Clostridium species infection is responsible for the fulminant progressing nature of the disease because the devitalized, ischemic liver parenchyma is ideal for such growth, and this is the first time that a such condition has been shown by CT images. Close observation with a high degree of suspicion is required for the successful treatment of such abscesses. PMID- 12734738 TI - Pseudo-Meigs' syndrome caused by ovarian metastasis from colon cancer: report of a case. AB - We report an extremely rare case of pseudo-Meigs' syndrome caused by ovarian metastases from colon cancer, and review the literature on this unusual entity. A 41-year-old woman was admitted for investigation of abdominal fullness and dyspnea. Preoperative examinations revealed a huge pelvic tumor, adenocarcinoma of the sigmoid colon, marked ascites, and bilateral pleural effusion. Laparotomy confirmed that the huge mass was comprised of bilateral ovarian tumors. Resection of the sigmoid colon and bilateral oophorectomies were performed. Although short term intrathoracic drainage was required, the hydrothorax and ascites rapidly resolved in the postoperative period. The patient died of disseminated liver and bone metastases 8 months after her operation; however, ascites and hydrothorax were not clinically noted until death. This and five other reported cases demonstrate that ovarian metastasis from colorectal cancer may occasionally cause pseudo-Meigs' syndrome, and that resection of the ovarian lesions could improve the prognosis. PMID- 12734740 TI - Solitary pancreatic metastasis from renal cell carcinoma concomitant with early gastric cancer 17 years after nephrectomy: report of a case. AB - The pancreas is an uncommon site for metastasis from renal cell carcinoma. We report the case of a 70-year-old man in whom a solitary pancreatic metastasis from renal cell carcinoma, found 17 years after nephrectomy, was successfully resected, combined with gastrectomy for early gastric cancer. We also discuss the relevant literature, including all the reports of pancreatic metastasis from renal cell carcinoma found in Medline. More than half the cases, like ours, were asymptomatic. A good prognosis can be expected once the pancreatic metastatic lesions are surgically excised, especially if it is a solitary metastasis. Therefore, surgical resection of pancreatic metastases is recommended to achieve the best chance of long-term survival. Special attention must be paid to the possibility of recurrence, even more than 10 years after nephrectomy for renal cell carcinoma, and imaging modalities should be part of the routine follow-up to detect metastases at an early stage. PMID- 12734741 TI - Pulmonary embolism in a living-related kidney transplantation donor. AB - Living-related renal transplantation is the optimal therapy for patients with end stage renal disease (ESRD). Normally, complications are rare in living-related donor nephrectomy. However, we experienced a case of pulmonary embolism (PE). The incidence of PE in living donor nephrectomy is rare, but the total incidence of PE in surgical operations has recently increased. The patient in the case reported here was diagnosed relatively early and recovered with appropriate treatment. It is very important for surgeons to realize that serious complications such as PE can develop in any case of living donor nephrectomy. PMID- 12734742 TI - Comparison of plain radiographs with CT scan to evaluate interbody fusion following the use of titanium interbody cages and transpedicular instrumentation. AB - The availability of lumbar interbody cages has fuelled renewed interest in interbody fusion. Despite this, there is no consensus regarding the best non invasive method for evaluation of interbody fusion, especially where cages have been used. The purpose of this study was to determine whether high-quality thin slice (1- to 3-mm) computed tomography (CT) scans allow proper evaluation of interbody fusion through titanium cages. Patients undergoing lumbar interbody fusion were prospectively evaluated with CT scan and plain radiographs 6 months following surgery. These images were blindly and independently evaluated by a consultant radiologist and a spine research fellow, for bridging bony trabeculation both through and surrounding the cages as well as for changes at the cage endplate interface. Fifty-three patients (156 cages) undergoing posterior lumbar interbody fusion using titanium interbody cages were evaluated. Posterior elements were used to pack the cages and no graft was packed outside the cages. The outcome data were analysed using the Kappa co-efficient and chi squared analysis. On CT scan, both observers noted bridging trabeculation in 95% of the cages (Kappa 0.85), while on radiographs this was present in only 4% (Kappa 0.74). Both observers also identified bridging trabeculation surrounding the cages on CT scan in 90% of cages (Kappa 0.82), while on the radiographs this was 8% (Kappa 0.86). Radiographs also failed to demonstrate all the loose cages. The results of the study show that high-quality CT scans show images suggesting bridging bony trabeculae following the use of titanium interbody cages. They also appear to show consistent bone outside the cages in spite of no bone graft having been used, and they appear to be better than plain radiographs in the early detection of cage loosening. PMID- 12734744 TI - The European phenology network. AB - The analysis of changes in the timing of life cycle-events of organisms (phenology) has been able to contribute significantly to the assessment of potential impacts of climate change on ecology. These phenological responses of species to changes in climate are likely to have significant relevance for socio economic issues such as agriculture, forestry and human health and have proven able to play a role in raising environmental awareness and education on climate change. This paper presents the European Phenology Network (EPN), which aims to increase the efficiency, added value and use of phenological monitoring and research, and to promote the practical use of phenological data in assessing the impact of global (climate) change and possible adaptation measures. The paper demonstrates that many disciplines have to deal with changes in the timing of life-cycle events in response to climate change and that many different user groups are involved. Furthermore, it shows how EPN addresses issues such as (1) raising public awareness and education, (2) the integration and co-operation of existing observing systems, (3) integration and access to phenological information and (4) communication. PMID- 12734743 TI - Clinical analysis of cervical radiculopathy causing deltoid paralysis. AB - In general, deltoid paralysis develops in patients with cervical disc herniation (CDH) or cervical spondylotic radiculopathy (CSR) at the level of C4/5, resulting in compression of the C5 nerve root. Therefore, little attention has been paid to CDH or CSR at other levels as the possible cause of deltoid paralysis. In addition, the surgical outcomes for deltoid paralysis have not been fully described. Fourteen patients with single-level CDH or CSR, who had undergone anterior cervical decompression and fusion for deltoid paralysis, were included in this study. The severity of deltoid paralysis was classified into five grades according to manual motor power test, and the severity of radiculopathy was recorded on a visual analog scale (zero to ten points). The degree of improvement in both the severity of deltoid paralysis and radiculopathy following surgery was evaluated. Of 14 patients, one had C3/4 CDH, four had C4/5 CDH, three had C4/5 CSR, one had C5/6 CDH, and five had C5/6 CSR. Both deltoid paralysis and radiculopathy improved significantly with surgery (2.57+/-0.51 grades vs 4.14+/ 0.66, P=0.001, and 7.64+/-1.65 points vs 3.21+/-0.58, P=0.001, respectively). In conclusion, the current study demonstrates that deltoid paralysis can develop due to CDH or CSR not only C4/5, but also at the levels of C3/4 and C5/6, and that surgical decompression significantly improves the degree of deltoid paralysis due to cervical radiculopathy. PMID- 12734745 TI - Bioavailability of a pediatric amlodipine suspension. AB - Currently, a suspension of crushed tablets of amlodidpine is widely used in children with hypertension without knowledge of its bioavailability. A comparative bioavailability study of a tablet and suspension formulation of amlodipine was completed in 20 healthy adult volunteers. Bioequivalence of the suspension was not different from the tablets. These results support the use of the suspension in children who cannot take the tablet. PMID- 12734746 TI - Body proportions before and during growth hormone therapy in children with chronic renal failure. AB - Growth retardation is a common problem in children with chronic renal failure (CRF). Few published data are available on whether the normalization of height in these children during growth hormone (GH) treatment is accompanied by proportional growth of the other parts of the body. In this study, body proportions before and during GH therapy were assessed in children with severe growth retardation due to CRF. Various body segments, such as sitting height, arm span, tibia, hand and foot length, biacromial and biiliacal diameter were measured in 15 children participating in a double-blind placebo-controlled cross over trial and in 22 children participating in a double-blind dose-response trial. Twelve children continued GH therapy after having participated in one of the two former trials and received GH therapy for 4 years. All results were adjusted for age and sex, and expressed as SD scores using reference values for healthy Dutch children. To assess body proportions, the various body segments were related to height and expressed as shape values (SV). At baseline all body segments SD scores were significantly lower than zero, indicating that the stunted growth of children with CRF included all body segments. Since height was not significantly more or less affected than the other body segments, all children had normal SV, indicating normal body proportions. The placebo controlled study showed a significant increase of the SD scores of height and several body segments during 6 months of GH [28 IU/m(2) per week (or 1.3 mg/m(2) per day)] versus placebo. The dose-response study demonstrated that height SDS as well as all other body segments SD scores increased significantly during 2 years of GH therapy with 28 IU/m(2) per week, compared with treatment with 14 IU/m(2) per week. Also during 4 years of GH therapy with 28 IU/m(2) per week, body segment SD scores increased to the same extent as height SDS, showing that GH did not significantly change SV i.e., body proportions. Both before and during GH therapy, children on dialysis had normal body proportions, comparable with children on conservative renal treatment. In conclusion, children with severe growth retardation due to CRF maintain normal body proportions in spite of their chronic disease. GH therapy with 28 IU/m(2) per week induces and maintains catch up growth of height and all body segments without signs of disproportionate growth. Thus GH therapy does not negatively influence body proportions in children with severe growth retardation secondary to CRF. PMID- 12734747 TI - Effect of lisinopril on proteinuria in children with nephrotic syndrome in Ilorin, Nigeria. PMID- 12734749 TI - Neonatal nephrocalcinosis in association with glucose-galactose malabsorption. AB - We report a case of severe nephrocalcinosis related to hypercalcaemia in a newborn with glucose-galactose malabsorption. He presented with poor growth and was noted to have polyuria, which was later recognised to be severe watery diarrhoea. We discuss the possible aetiological factors for nephrocalcinosis in this condition. PMID- 12734748 TI - Thyroxine prevents reoxygenation injury in isolated proximal tubule cells. AB - Ischemia is characterized by cessation of blood flow and oxygen delivery to tissues that results in disruption of cellular structure and organelles. However, restoration of blood flow following ischemia causes reperfusion injury, characterized by further damage in the tissues mediated by reactive oxygen species. In the kidney, reactive oxygen molecules have been implicated in ischemic, toxic and immunological glomerular damage. Thyroxine has been shown to be cytoprotective in toxic and ischemic injury. Thyroxine's cytoprotective effect is postulated to be secondary to stimulation of intracellular ATP synthesis. However, the underlying mechanism of that beneficial effect remains to be investigated. In this study we investigated the effect of thyroxine (T4) on free oxygen radical production in an in vitro model of reperfusion injury. Free oxygen radical (FOR) levels were determined by a chemiluminescence method after freshly isolated rabbit proximal tubule cells were subjected to 15 min of anoxia followed by 45 min of reoxygenation. Reoxygenation injury resulted in a significant increase in FOR levels (P<0.0001). FOR levels were significantly lower in the group treated with thyroxine (P=0.01) and cells treated with thyroxine displayed better preservation of cellular structure. We conclude that thyroxine's cytoprotective effect might be via decreased synthesis of FOR, and thyroxine treatment may confer cytoprotection in renal conditions characterized by FOR mediated injury. PMID- 12734751 TI - cAMP signaling in Aspergillus fumigatus is involved in the regulation of the virulence gene pksP and in defense against killing by macrophages. AB - Aspergillus fumigatus is an important pathogen of immunocompromised hosts, causing pneumonia and invasive disseminated disease and resulting in high mortality. In order to determine the importance of the cAMP signaling pathway for virulence, three genes encoding putative elements of the pathway have been cloned and characterized: the adenylate cyclase gene acyA, and gpaA and gpaB, both of which encode alpha subunits of heterotrimeric G proteins. The acyA and gpaB genes were each deleted in A. fumigatus. Both mutants showed reduced conidiation, with the deltaacyA mutant producing very few conidia. The growth rate of the deltaacyA mutant was also reduced, in contrast to that of the deltagpaB mutant. Addition of 10 mM dibutyryl-cAMP to the culture medium completely restored the wild-type phenotype in both mutant strains. To study the influence of GPAB on the expression of the gene pksP, which encodes a virulence factor that is involved in pathogenicity, a pksPp-lacZ gene fusion was generated and integrated as a single copy at the pyrG gene locus of both the parental strain and the deltagpaB mutant strain. The deltagpaB mutant showed reduced expression of the pksPp-lacZ reporter gene relative to that in the parental strain. In mycelia of both the parental strain and the deltagpaB mutant pksPp-lacZ expression was increased when isobutyl methyl-xanthine, an inhibitor of intracellular phosphodiesterases, was added to the medium. The survival rate of conidia after ingestion by human monocyte derived macrophages was also determined. The killing rate for conidia from deltaacyA and deltagpaB strains was significantly higher than that for wild-type conidia. Taken together, these findings suggest that cAMP triggers a system that protects A. fumigatus from the effects of immune effector cells of the host. PMID- 12734750 TI - Heterologous transposition in Ustilago maydis. AB - The phytopathogenic basidiomycete Ustilago maydis has become a model system for the analysis of plant-pathogen interactions. The genome sequence of this organism will soon be available, increasing the need for techniques to analyse gene function on a broad basis. We describe a heterologous transposition system for U. maydis that is based on the Caenorhabditis transposon Tc1, which is known to function independently of host factors and to be active in evolutionarily distant species. We have established a nitrate reductase based two-component counterselection system to screen for Tc1 transposition. The element was shown to be functional and transposed to several different locations in the genome of U. maydis. The insertion pattern observed was consistent with the proposed general mechanism of Tc1/mariner integration and constitutes a proof of principle for the first heterologous transposition system in a basidiomycete species. By mapping the insertion site context to known genomic sequences, Tc1 insertion events were shown to occur on different chromosomes, but exhibit a preference for non-coding regions. Only 20% of the insertions were found in putative open reading frames. The establishment of this system will permit efficient gene tagging in U. maydis and possibly also in other fungi. PMID- 12734752 TI - Combinatorial chemistry as a new approach in antiparasitic drug discovery. AB - Key challenges in antiparasitic drug discovery are target selection and identification of appropriate small molecules as potential ligands for these targets. Novel tools and techniques continue to be developed to address both of these problems. This report focuses on the application of a suite of technologies summarized as combinatorial chemistry. Recent success using these technologies is explored, reasons for frequent failure are given and discussed. PMID- 12734754 TI - Pleomorph poorly differentiated endocrine carcinoma of the rectum. AB - We present a case of poorly differentiated endocrine carcinoma (PDEC) of the rectum identified immunohistochemically and characterized by a high degree of cellular pleomorphism, including bizarre giant cells. This case indicates that gastrointestinal PDECs are not restricted to small cell carcinomas. Among the multiple genes investigated, loss of heterozygosity (LOH) of the p53 locus without p53 immohistochemical accumulation, overexpression of c-kit and absent expression of p16 were seen. PMID- 12734755 TI - Rearrangement of the genes for the biosynthesis of benzoxazinones in the evolution of Triticeae species. AB - Gramineous plants, including the major agricultural crops wheat (Triticum aestivum L.), rye (Secale cereale L.) and maize (Zea mays L.), accumulate benzoxazinones (Bxs) as defensive compounds. Previously, we isolated cDNAs of the Bx biosynthetic genes in wheat, TaBx2- TaBx5, that encode the enzymes catalyzing the sequential hydroxylation of indole to Bxs. In this study we isolated a cDNA of TaBx1, which encodes the first enzyme of the Bx pathway of wheat. The level of identity (80%) in deduced amino-acid sequence between TaBx1 and the corresponding maize gene Bx1 was as high as those shown between TaBx2- TaBx5 and the corresponding maize genes Bx2- Bx5, respectively. Southern blot analysis using the TaBx1- TaBx5 cDNAs as probes was conducted with aneuploid lines of hexaploid wheat in order to determine sub-chromosomal locations of the five Bx biosynthetic genes in Triticeae species. In wheat, TaBx1 and TaBx2 co-existed in specific regions of chromosomes 4A, 4B and 4D, and TaBx3- TaBx5 were localized together in the distal regions of the short arms of chromosomes 5A, 5B and 5D. TaBx3 and TaBx5 were found to have duplicated loci in the long arm and the short arm of chromosome 5B, respectively. In rye, homoeoloci of TaBx1 and TaBx2 were located on chromosome 7R and those for TaBx3- TaBx5 were located on chromosome 5R. In barley, no Southern blot band was detected with any of the probes under the highly stringent hybridization conditions, suggesting that the non-Bx phenotype of barley is attributable to the loss of Bx biosynthetic genes. PMID- 12734753 TI - Allelic imbalance regions on chromosomes 8p, 17p and 19p related to metastasis of hepatocellular carcinoma: comparison between matched primary and metastatic lesions in 22 patients by genome-wide microsatellite analysis. AB - To understand the molecular mechanisms of metastasis in hepatocellular carcinoma (HCC), it is necessary to identify the accumulating genetic alterations during its progression as well as those responsible for the acquisition of metastatic potential in cancer cells. In our previous study, using comparative genomic hybridization (CGH), we found that loss on chromosome 8p is more frequent in metastatic lesions than in matched primary tumors of HCC. Thus, 8p deletion might contribute to HCC metastasis. To narrow the location of metastasis-related alteration regions, we analyzed 22 primary and matched metastatic lesions of HCC by genome-wide microsatellite analysis. Common regions with high levels of allelic imbalance (AI) were identified on 17p, 8p11-cen, 8p21-23, 4q32-qter, 4q13 23, 16q, and 1p33. Regions with increased AI in metastatic lesions were 8p23.3, 8p11.2, 17p11.2-13.3, 4q21-22, 4q32-qter, 8q24.1, 9p11, 9q31, 11q23.1, 13q14.1 31, 13q32-qter, 16p13.3, 16q13, 16q22, and 19p13.1, and these were considered to be related to the metastasis phenotype. Among them, loss on 8p was again proved to be related to progression and metastasis of HCC, and 8p23.3 and 8p11.2 were two likely regions harboring metastasis-related genes. It was also shown for the first time in HCC that AI of 19p13.1 might also be related to metastatic potential. These results provide some candidate regions for further study to identify putative genes suppressing metastasis of HCC. PMID- 12734756 TI - Differential Al resistance and citrate secretion in barley (Hordeum vulgare L.). AB - While barley ( Hordeum vulgare L.) is the most sensitive species to Al toxicity among small-grain crops, variation in Al resistance between cultivars does exist. We examined the mechanism responsible for differential Al resistance in 21 barley varieties. Citrate was secreted from the roots in response to Al stress. A positive correlation between citrate secretion and Al resistance [(root elongation with Al)/(root elongation without Al)] and a negative correlation between citrate secretion and Al content of root apices, were obtained, suggesting that citrate secretion from the root apices plays an important role in excluding Al and thereby detoxifying Al. The Al-induced secretion of citrate was characterized using an Al-resistant variety (Sigurdkorn) and an Al-sensitive variety (Kearney). In Sigurdkorn, Al-induced secretion of citrate occurred within 20 min, and the secretion did not increase with increasing external Al concentration. The Al-induced citrate secretion ceased at low temperature (6 degrees C) and was inhibited by anion-channel inhibitors. Internal citrate content of root apices was increased by Al exposure in Sigurdkorn, but was not affected in Kearney. The activity of citrate synthase was unaffected by Al in both Al-resistant and Al-sensitive varieties. The secretion rate of organic acid anions from barley was the lowest among wheat, rye and triticale. PMID- 12734757 TI - The sodium/calcium exchanger family-SLC8. AB - The Na(+)/Ca(2+) exchanger gene family encompasses three distinct proteins, NCX1, NCX2, and NCX3, which mediate cellular Ca(2+) efflux and thus contribute to intracellular Ca(2+) homeostasis. NCX1 is expressed ubiquitously while NCX2 and NCX3 are limited to brain and skeletal muscle. NCX1 exchanges 3 extracellular Na(+) for 1 intracellular Ca(2+). In addition to transporting Na(+) and Ca(2+), NCX1 activity is also regulated by these cations. NCX1 is especially important in regulating cardiac contractility. PMID- 12734758 TI - Influence of prolonged intermittent high-intensity exercise on knee flexor strength in male and female soccer players. AB - This study investigated the effect of an acute, prolonged, intermittent, high intensity single-leg pedalling exercise task (PIHIET) on the isokinetic leg strength of the knee flexors in six male and seven female collegiate soccer players. Following determination of single-leg VO(2peak), subjects completed a PIHIET designed to simulate the energetics of soccer match play (approximately 90 min in total; approximately 70% single-leg VO(2peak)). Pre-, mid- and post-PIHIET gravity-corrected indices of knee flexion peak torque (PT) and range of motion relativised torque at 15% of knee flexion (RRT(15%); 0% = full knee extension) were assessed at a lever-arm angular velocity of 1.05 rad.s(-1)for intervention and control limbs using an isokinetic dynamometer. Repeated measures ANOVAs revealed significant condition (PIHIET, control) x time (pre-, mid-, post-PIHIET) interactions for knee flexion PT (F([2,22])=26.2; P<0.001) and RRT(15%) (F([2,22])=20.1; P<0.001). Flexion PT and RRT(15% )were observed to decrease, pre to post-intervention, from 92.8 (28.7) N.m to 72.1 (28.0) N.m and from 63.8 (17.5) N.m to 47.9 (18.4) N.m respectively, for the intervention limb alone. These data corresponded to 22.3% and 24.9% mean reductions pre-post intervention in PT and RRT(15%). Exploratory post hoc analysis of the pattern of the relative deterioration (%) of PT and RRT(15%), for the intervention limb alone, revealed a three-way interaction [group (male, female) x parameter (PT, RRT(15%)) x assessment phase (pre- to mid-PIHIET, mid- to post-PIHIET)] (F(1,11)=5.2; P<0.05). This interaction characterised a greater deterioration of strength performance during the mid- to post-PIHIET assessment phase, at the extremes of range of motion (RRT(15%)) for the female group. The greater percentage of mid post phase strength loss observed in women near the end-range extension may potentially be implicated in the higher incidence of knee injury reported in female soccer players. PMID- 12734759 TI - Muscle hypertrophy, hormonal adaptations and strength development during strength training in strength-trained and untrained men. AB - Hormonal and neuromuscular adaptations to strength training were studied in eight male strength athletes (SA) and eight non-strength athletes (NA). The experimental design comprised a 21-week strength-training period. Basal hormonal concentrations of serum total testosterone (T), free testosterone (FT) and cortisol (C) and maximal isometric strength, right leg 1 repetition maximum (RM) of the leg extensors were measured at weeks 0, 7, 14 and 21. Muscle cross sectional area (CSA) of the quadriceps femoris was measured by magnetic resonance imaging (MRI) at weeks 0 and 21. In addition, the acute heavy resistance exercises (AHRE) (bilateral leg extension, five sets of ten RM, with a 2-min rest between sets) including blood samples for the determination of serum T, FT, C, and GH concentrations were assessed before and after the 21-week training. Significant increases of 20.9% in maximal force and of 5.6% in muscle CSA in NA during the 21-week strength training period were greater than those of 3.9% and 1.8% in SA, respectively. There were no significant changes in serum basal hormone concentrations during the 21-week experiment. AHRE led to significant acute decreases in isometric force and acute increases in serum hormones both at weeks 0 and 21. Basal T concentrations (mean of 0, 7, 14 and 21 weeks) and changes in isometric force after the 21-week period correlated with each other (r=0.84, P<0.01) in SA. The individual changes in the acute T responses between weeks 0 and 21 and the changes in muscle CSA during the 21-week training correlated with each other (r=0.76, P<0.05) in NA. The correlations between T and the changes in isometric strength and in muscle CSA suggest that both serum basal testosterone concentrations and training-induced changes in acute testosterone responses may be important factors for strength development and muscle hypertrophy. PMID- 12734760 TI - Performance for short intermittent runs: active recovery vs. passive recovery. AB - The purpose of this study was to compare the effects of active vs. passive recovery on the time to exhaustion for intermittent runs (15 s) at supramaximal velocity (120% of maximal aerobic speed). Twelve male subjects performed a graded test, an intermittent run to exhaustion with active recovery (50% of maximal aerobic speed) and an intermittent run to exhaustion with passive recovery. Results showed that intermittent runs to exhaustion with passive recovery [745 (171) s] allowed subjects to run for a significantly longer (p<0.001) time than intermittent runs to exhaustion with active recovery [445 (79) s]. These results could be explained by a significantly higher (p<0.001) energy requirement for intermittent runs with active recovery [59.9 (9.6) ml.kg(-1).min(-1)] than for intermittent runs with passive recovery [48.9 (6.9) ml.kg(-1).min(-1)]. It could be also hypothesized that the energy required to run during short active recovery would result in less oxygen being available to reload myoglobin and haemoglobin, to remove lactate concentrations and to resynthesize the phosphocreatine. Consequently, for intermittent runs with short recovery periods, passive recovery will induce a longer time to exhaustion than active recovery. PMID- 12734761 TI - Long-term treatment of schizoaffective disorder: review and recommendations. AB - OBJECTIVE: To provide an overview of long-term treatment studies in schizoaffective disorder (SAD) and to draw conclusions for clinical decision making. METHOD: Literature was identified by searches in Medline, Embase, and the Cochrane Controlled Trials Register as well as a hand-search of handbook and journal articles. Studies were considered relevant if they reported on trials of at least 6 months duration and if they presented data for the SAD patients in particular. RESULTS: Thirty-nine studies met the criteria and 18 used modern diagnostic criteria, i. e., RDC, DSM-III-R, -IV, or ICD-10. The studies focused on lithium, anticonvulsants, and antipsychotics. The scientific evidence for prophylactic efficacy of the different substances is poor. Nevertheless, the data encourage the use of lithium and carbamazepine in primarily affective patients and clozapine in primarily schizophrenic patients and possibly in mainly affective patients as well. CONCLUSIONS: There is a considerable need for prospective and controlled studies on the long-term treatment of SAD. However, it seems to be useful to subtype the disorder of the patients into primarily affective vs. schizophrenic schizoaffective disorder and schizodepressive vs. schizobipolar and to treat accordingly. PMID- 12734762 TI - Current perception thresholds of patients with long-term administration of maprotiline. AB - The purpose of this study was to evaluate sensory nerve function using current perception thresholds (CPTs) in patients who were administrated maprotiline. Twelve patients with post-herpetic neuralgia and 20 control subjects were studied. The patients with post-herpetic neuralgia were given a daily dose of 60 mg of maprotiline and were maintained at the same dose for 6 months. Twenty control subjects were randomly selected from healthy volunteers. ACPT test was used for quantitative assessment of A beta, A sigma, and C fiber transmission, which are associated with pain, by three (2000, 250, and 5 Hz) different frequencies of electric stimulation. CPTs of 5, 250, and 2000 Hz in patients with post-herpetic neuralgia 2 months after administration of maprotiline were 141.7 +/- 17.3 for 5 Hz, 120.8 +/- 12.9 for 250 Hz, and 256.4 +/- 18.0 for 2000 Hz, which were significantly (P < 0.01) higher than those (67.0 +/- 9.1 for 5 Hz, 73.4 +/- 7.0 for 250 Hz, and 191.3 +/- 20.2 for 2000 Hz) before treatment and than those (35.3 +/- 15.8, 62.0 +/- 18.9, and 198.9 +/- 15.8) of control subjects. An increase in CPT for 5 Hz at 2 months after administration of maprotiline correlated (r = 0.71, p = 0.01) with a decrease in pain score. There were no correlations between an increase in CPT and changes in Hamilton Depression Scale (HAMD) values until 3 months after maprotiline treatment. However, we found that an increase in CPT for 5 Hz at 6 months after maprotiline treatment correlated (r = 0.68, p = 0.015) with a decrease in HAMD values. In conclusion, administration of 60 mg maprotiline significantly increased current perception thresholds at 2 months after the administration. PMID- 12734763 TI - Antipsychotic efficacy of the antidepressant trimipramine: a randomized, double blind comparison with the phenothiazine perazine. AB - BACKGROUND: The tricyclic antidepressant trimipramine exhibits several features (e. g., dopaminergic effect, molecular structure similar to a neuroleptic, receptor-binding profile similar to clozapine) that suggest its potential as an antipsychotic medication. The aim of the study was to investigate the antipsychotic potential of trimipramine in a controlled clinical trial comparing its antipsychotic efficacy with that of a neuroleptic. METHOD: In a German multi center, randomized, double-blind trial, the antipsychotic efficacy of trimipramine was compared with that of the phenothiazine neuroleptic perazine, using the Brief Psychiatric Rating Scale (BPRS), the Positive and Negative Syndrome Scale (PANSS), and Clinical Global Impressions (CGI). Antidepressant efficacy of both agents was measured by use of the Bech-Rafaelsen Melancholia Scale (BRMES). Ninety-five patients with acute schizophrenia (DSM-III-R) and a BPRS total score > 40 at baseline were treated with either 300-400 mg trimipramine or 450-600 mg perazine for 5 weeks. RESULTS: Therapeutic equivalence of both treatments (in the dosages used) could not be demonstrated (change in BPRS total score, per-protocol [PP] analysis, one-sided equivalence testing). However, intention-to-treat (ITT) as well as PP analysis showed a statistically significant decrease in the BPRS total scores in both treatment groups (PP: trimipramine, 56.5 +/- 9.8 to 44.1 +/- 17.9; perazine, 56.4 +/- 10.8 to 37.9 +/- 12.9). Significant decreases in all BPRS and PANSS subscores as well as CGI results and response rate support the antipsychotic efficacy of trimipramine. The BRMES total scores significantly decreased in both treatment groups without showing a significant difference between the two agents. Trimipramine was better tolerated than perazine and did not elicit extrapyramidal symptoms. CONCLUSION: Trimipramine failed to exhibit therapeutic equivalence to perazine in the dosages used. However, there was evidence of a substantial antipsychotic effect of trimipramine. It may be a useful medication if depressive symptoms in psychotic patients require antidepressant treatment or if other antipsychotics cannot be administered. PMID- 12734764 TI - Transient restless legs-like syndrome as a complication of opiate withrawal. AB - In most cases, restless legs syndrome (RLS) is an idiopathic disorder affecting patients of middle and older age. Usually, the intensity of symptoms is constant or progressive. Here, we describe two opiate addicts developing RLS during opiate withdrawal. Their RLS disappeared along with the remission of withdrawal symptoms. Both cases responded well to a treatment with Madopar (L-dopa plus benserazid). This observation encouraged us to perform a retrospective analysis of all cases of opiate addicts detoxified in our unit during a 5-month period. This analysis revealed 15 out of 120 patients who had described the symptoms of transient RLS emerging during opiate detoxification treatment. Transient RLS during opiate detoxification merits further interst in order to improve the treatment of sleep disturbances during detoxification and as a model of interaction of the dopaminergic and endorphine system in motor activity. PMID- 12734765 TI - CYP2D6 polymorphism and tardive dyskinesia in schizophrenic patients. AB - Antipsychotic drug-induced tardive dyskinesia (TD) is a serious problem during psychopharmacologic treatment of schizophrenic patients. In search of genetic factors contributing to TD, there is a lack of consensus regarding the role of the polymorphic isozyme cytochrome P450 CYP2D6, which is involved in the oxidative metabolism of antipsychotic drugs. In the present case-control study, we tested the putative influence of the CYP2D6 genotype on the development of TD. Out of 157 patients, 109 were retrospectively selected meeting DSM IV criteria for schizophrenia or schizoaffective disorder, and 50 of them persistently presenting with TD. Genotyping detected the functional allele CYP2D6 *1, the known major defective alleles CYP2D6 *3, *4, *5, *6, and gene duplication. According to their number of functional CYP2D6 alleles, subjects were divided into carriers of none, one, or at least two functional CYP2D6 alleles. The proportions of these categories did not differ between patients and an ethnically homogenous control population (n = 195, p = 0.99) or between patients with and without TD (p = 0.818). Schizophrenic patients were carriers of gene duplication more often than healthy probands, without revealing statistical significance (p = 0.10). Out of seven patients with gene duplication, three developed persistent TD. Furthermore, patients with and without TD were comparable according to age, age of onset, gender, and duration of illness, but subjects with TD had taken more lifetime chlorpromazine equivalents (CPZ) than had patients without TD (chi 2-test, Student's t-test). Forward as well as backward logistic regression analyses confirmed that the presence of TD was influenced by lifetime CPZ but not by age, age of onset, gender, duration of illness, or CYP2D6 genotype. In contrast to the relevance of lifetime CPZ, the lifetime dose of antipsychotic drugs known to be metabolized by CYP2D6 did not significantly influence the presence of TD. In conclusion, our results provide no evidence for the contribution of CYP2D6 genotype to the development of TD in schizophrenic patients receiving long-term antipsychotic medication. PMID- 12734767 TI - Uncommon side effects associated with olanzapine. A case report. PMID- 12734766 TI - Methylphenidate enhances both intracortical inhibition and facilitation in healthy adults. AB - Transcranial magnetic stimulation was used to investigate the effect of the psychostimulant drug methylphenidate (MPH) on motor cortex excitability in healthy adults (n = 12) in a placebo-controlled, crossover design study. MPH caused an enhancement of intracortical inhibition as well as intracortical facilitation. Enhancement of both of these TMS parameters was unexpected and suggests that MPH exerts its action on the motor cortex not only through the dopaminergic neurotransmitter system. PMID- 12734768 TI - Successful antimanic treatment and mood stabilization with lamotrigine, clozapine, and valproate in a bipolar patient after lithium-induced cerebellar deterioration. A case report. PMID- 12734769 TI - Cholesterol, essential fatty acids, and suicide. PMID- 12734771 TI - AIDS-related lipodystrophy/insulin resistance syndrome. AB - The recent development and clinical use of three different types of highly effective anti-HIV-1 drugs, including nucleotide and non-nucleotide reverse transcriptase inhibitors (NRTIs) and non-peptidic viral protease inhibitors (PIs) and their combinations, termed highly active antiretroviral therapy (HAART), have dramatically reduced the infection-related mortality of AIDS patients in developed countries. However, the prolongation of the life expectancy of HIV-1 infected patients and/or long-term use of the above antiviral agents have generated a score of new problems and complications. Among them is the relatively common AIDS-related lipodystrophy/insulin resistance syndrome, which is associated with severe metabolic disturbances such as carbohydrate intolerance/diabetes mellitus and severe dyslipidemia, which influence the quality of life and threaten the life expectancies of HIV-1-infected patients by increasing the risk of atherosclerotic cardiovascular disease. The etiology of this syndrome appears to be multi-factorial; the classes of anti-viral drugs listed above, hypercytokinemia in AIDS patients, and the HIV-1 infection itself could induce the pathologic changes of this syndrome or increase the vulnerability of patients to the adverse effect of the therapeutic compounds. In this article, we review our current understanding of the pathogenesis of this severe AIDS-associated metabolic disorder. PMID- 12734772 TI - Effects of estradiol and progesterone on vertebral collagen, glycosaminoglycans and phosphatases in ovariectomized adult rats. AB - Vertebral collagen, glycosaminoglycans (GAGs), tartrate-resistant acid phosphatase (TRAP) and alkaline phosphatase (ALP) were measured in ovariectomized (ovx) adult Wistar rats treated with estradiol (E 2 ) (10 micro g/kg BW for 35 days on alternate days, and progesterone (P 4 ) (140 micro g/kg BW for 35 days on alternate days) in E 2 + P 4 treated rats. P 4 given alone or in combination with E 2 significantly increased the levels of collagen in the vertebral bone. Neither ovx nor E 2 treatment altered the concentration of collagen in these rats. Administration of E 2 or P 4 significantly decreased the concentration of hyaluronic acid (HA), but remaining unaffected when a combination of these steroids was given. In contrast to their effect on HA, E 2 and P 4 each significantly increased the levels of chondroitin sulfate (CS) in the vertebral bone. The specific activity of ALP was decreased after ovx. E 2 and P 4 alone or in combination also registered a significant decrease in the activities of ALP and TRAP. The results suggest that E 2 and P 4 each exert definite effects on vertebral bone turnover in ovariectomized rats. PMID- 12734773 TI - Estrogen reduces pro-inflammatory cytokines in rodent adipose tissue: studies in vivo and in vitro. AB - Obesity is associated with an increased risk of developing insulin resistance, Type 2 diabetes, and cardiovascular disease. Reports have suggested that adipose tissue-derived cytokines such as tumor necrosis factor-alpha, interleukin-6 and interleukin-8 could be involved in the development of these health complications. Since estrogen has been suggested to attenuate the development of atherosclerosis and cardiovascular disease, we investigated whether ovariectomy affected the production and release of these three adipose tissue-derived cytokines with and without estrogen replacement in vivo and in vitro. Female Wistar rats were submitted to either a) ovariectomy, b) ovariectomy and estrogen replacement, or c) sham operation. After five months, animals were sacrificed and parametrial adipose tissue was removed and incubated for up to 24 hours with either interleukin-1beta (IL-1beta) (5 micro g/l), dexamethasone (50 nM) or estrogen (50 nM). Ovariectomy significantly increased interleukin-6 gene expression (p < 0.05) as well as interleukin-8 protein levels (p < 0.05) and gene expression (p < 0.05) in the adipose tissue, and estrogen replacement significantly reversed this increase (p < 0.05). However, no direct effects of estrogen were found in in vitro adipose tissue incubations. Neither ovariectomy nor estrogen replacement had any effects on tumor necrosis factor-alpha protein levels or gene expression. In conclusion, estrogen-deficient rats were found to have increased production of interleukin-6 and interleukin-8, which could be attenuated by estrogen replacement. Since estrogen is suggested to be anti-atherosclerotic, this effect might be caused by a reduction in cytokine production from the adipose tissue. PMID- 12734774 TI - Interleukin (IL)-6 mRNA expression is stimulated by insulin, isoproterenol, tumour necrosis factor alpha, growth hormone, and IL-6 in 3T3-L1 adipocytes. AB - Interleukin (IL)-6 has recently been shown to be an adipocyte-expressed cytokine. Its serum concentrations are elevated in insulin resistance and obesity. For further evaluation of IL-6 gene expression regulation, fully differentiated 3T3 L1 adipocytes were treated with various hormones known to induce insulin resistance. IL-6 mRNA content was measured by quantitative real-time reverse transcription-polymerase chain reaction. Interestingly, treatment of adipocytes with 100 nM insulin, 10 micro M isoproterenol, 10 ng/ml tumour necrosis factor alpha (TNFalpha), and 500 ng/ml growth hormone (GH) for 16 h stimulated IL-6 mRNA expression 2.3-fold, 47-fold, 74-fold, and 1.4-fold, respectively (p < 0.01). In contrast, treatment with 100 nM dexamethasone significantly decreased IL-6 expression to 32 % of control levels (p < 0.01), whereas triiodothyronine and angiotensin 2 did not have any effect. Furthermore, stimulation of IL-6 expression was time-dependent with maximal stimulatory effects detectable after 1 h of insulin, isoproterenol, and GH addition and 12 h of TNFalpha, respectively. Moreover, isoproterenol's effect could be almost completely reversed by pretreatment of 3T3-L1 cells with the beta-adrenergic antagonist propranolol and mimicked by stimulation of G S -proteins with cholera toxin and adenylyl cyclase with forskolin and dibutyryl cAMP, respectively. Finally, IL-6 strongly induced its own expression in a time-dependent fashion. Taken together, our results demonstrate that IL-6 expression in adipocytes is governed by an autocrine positive feedback loop and upregulated by insulin, isoproterenol, TNFalpha, and GH. In concert with this adipocytokine's upregulation in states of decreased insulin sensitivity such as obesity and diabetes, the data support a possible role of IL-6 as a selectively regulated mediator of insulin resistance. PMID- 12734775 TI - Gonadotropin-releasing hormone increases melatonin release in the pineal gland of the female rat in vitro. AB - To evaluate the effect of gonadotropin-releasing hormone (GnRH) on melatonin ( N acetyl-5-methoxytryptamine) release and its synthesizing enzyme activities in pineal glands, pineals from adult female rats during diestrus were organ-cultured in a medium containing 10 -12, 10 -10, or 10 -8 M GnRH for 6 h. Melatonin release increased significantly in pineals cultured with 10 -10 and 10 -8 M GnRH compared to controls. However, in pineal glands that were organ-cultured in a medium containing 10 -12 to 10 -8 M GnRH, the activity of arylalkylamine N acetyltransferase, which is the key regulatory enzyme in melatonin biosynthesis, showed no significant difference from controls. Likewise, GnRH at these concentrations had no significant effect on the activity of pineal hydroxyindole- O-methyltransferase, which catalyzes the final step of melatonin biosynthesis. These results show that GnRH stimulates pineal melatonin release, but suggest that GnRH does not affect its melatonin synthesis. PMID- 12734777 TI - Effect of losartan on insulin plasma concentrations and LPL activity in adipose tissue of hypertensive rats. AB - The aim of this study is to determine the effect of losartan on insulin and angiotensin II (Ang II) concentrations in plasma as well as on lipoprotein lipase activity (LPL) and angiotensin II content in the adipose tissue of hypertensive rats. Fifty male rats were divided in five groups. Group A served as controls. Group B underwent renal artery stenosis. Group C were administered losartan (10 mg/kg/day) per os, while rats in group D were submitted to renal artery stenosis and were treated with losartan as above. Group E was used as sham-operated control. The animals were sacrificed at day 21. Blood samples were collected, and perirenal adipose tissue was isolated. Furthermore, adrenal's were removed and their relative weight (adrenal weight/body weight) was used as an index of sympathetic stimulation. According to our results, renovascular hypertension resulted in lower insulin concentrations and higher Ang II content in plasma. In hypertensive rats, LPL activity was decreased, while the adrenals' relative weight was elevated. On the other hand, losartan administration resulted in normalization of insulin concentrations in plasma and adrenals' relative weight, with consequent up regulation of LPL activity in adipose tissue. In conclusion, renovascular hypertension interferes in lipid metabolism by reducing LPL activity in adipose tissue, while losartan administration reverses this effect by enhancing insulin release and reducing sympathetic nervous system (SNS) stimulation. PMID- 12734776 TI - Culture of human adipose tissue explants leads to profound alteration of adipocyte gene expression. AB - Primary culture of adipose tissue has often been used to investigate pharmacological and nutritional regulation of adipocyte gene expression. Possible alteration of adipocyte gene expression by primary culture on its own has not been explored in detail. In order to address this issue, explants were prepared from human subcutaneous adipose tissue recovered from plastic surgery and maintained for 0 to 48 h in DMEM supplemented with 10 % serum. At different time points, adipocytes were isolated from the explants by collagenase digestion, and mRNA expression and lipolysis were studied. Culture was associated with an accumulation of tumor necrosis factor-alpha (TNFalpha) in the culture medium, an increase in anaerobic glycolysis, and an increase in the basal lipolysis. In parallel, a rapid and dramatic decrease in the level of mRNA encoding for several adipocyte-specific proteins such as adipocyte lipid-binding protein, hormone sensitive lipase, lipoprotein lipase, and peroxisome proliferation activating receptor-gamma2 was observed in isolated adipocytes. These downregulations were reminiscent of a dedifferentiation process. In parallel, primary culture was associated with an increase in adipocyte beta-actin, TNFalpha, glucose transporter-1 and hypoxia-induced factor-1alpha mRNAs. Treatment of explants with agents that increase cAMP (isobutylmethylxanthine and forskolin) prevented TNFalpha production and expression and culture-induced alterations of adipocyte gene expression. These data show that primary culture of human adipose tissue explants dramatically alters adipocyte gene expression. PMID- 12734779 TI - Differences in plasminogen activator inhibitor 1 in subcutaneous versus omental adipose tissue in non-obese and obese subjects. AB - Human adipose tissue can produce plasminogen activator inhibitor-1 (PAI-1). It has been suggested that high levels of PAI-1 are of importance in enhanced cardiovascular disease observed among obese subjects, especially abdominally obese individuals. In the present study, we investigated the level of mRNA and production of PAI-1 in adipose tissue from two adipose tissue depots (omental vs. subcutaneous). Adipose tissue from both depots was obtained from obese (mean BMI, 46.9 kg/m 2) and non-obese (mean BMI, 23.9 kg/m 2) women. PAI-1 mRNA was measured both in fresh adipose tissue obtained immediately after surgery and after the adipose tissue (fragments) had been incubated for up to 72 h. In immediately frozen adipose tissue, PAI-1 mRNA expression was similar in omental and subcutaneous adipose tissue. No differences between obese and non-obese women were found. However, when adipose tissue fragments were cultured, PAI-1 mRNA and PAI-1 production were significantly higher in omental than in subcutaneous adipose tissue (p < 0.05). In the culture system, the production of PAI-1 in obese subjects was higher than in non-obese subjects in both subcutaneous (p < 0.05) and in omental adipose tissue (p = 0.19). In order to test whether these regional differences observed after incubation of the adipose tissue were due to differences in local accumulation of cytokines that may stimulate PAI-1 by a paracrine or autocrine manner, we investigated the expression of transforming growth factor beta1 (TGF-beta1) mRNA and tumor necrosis factor alpha (TNF-alpha) mRNA and protein. No differences between the two fat depots were found. In conclusion, no differences in PAI-1 expression between omental and subcutaneous adipose tissue were observed in biopsies frozen immediately after removal, but after incubation of adipose tissue (which somehow stimulates PAI-1 production), higher levels of PAI-1 were found in omental adipose tissue than in subcutaneous adipose tissue. Finally, PAI-1 production in adipose tissue from obese women was higher in non-obese women after incubation for 72 h. PMID- 12734778 TI - SOCS-3 is involved in the downregulation of the acute insulin-like effects of growth hormone in rat adipocytes by inhibition of Jak2/IRS-1 signaling. AB - One of the long-term effects of growth hormone (GH) in adipocytes is to maintain a state of refractoriness to insulin-like effects, a refractoriness which otherwise declines within a few hours of GH starvation. Here, we examined differences in GH signaling and the possible role for the recently identified family of suppressors of cytokine signaling (SOCS) proteins in the transition between the refractory and the responsive states in rat adipocytes. The ability of GH to stimulate lipogenesis and tyrosine phosphorylation of the GH receptor (GHR), Janus kinase 2 (Jak2), insulin receptor substrate-1 (IRS-1) and -2 (IRS-2) was greatly reduced in refractory as compared to responsive primary rat adipocytes. However, phosphorylation of Signal Transducer and Activator of Transcription 5 (Stat5) was not affected. SOCS-3 and CIS mRNA levels were significantly higher in refractory compared to responsive cells and could be induced by GH, whereas the level of SOCS-2 mRNA was unchanged. With overexpression of GHR, Jak2 and IRS-1 along with each of these SOCS proteins in human A293 cells, we could demonstrate that both SOCS-1 and SOCS-3 completely inhibited the GH-stimulated tyrosine phosphorylation of IRS-1, whereas SOCS-2 and CIS did not. Our data suggest that GH induces refractoriness to the insulin-like effects in a negative-feedback manner by inhibiting GH-induced GHR/Jak2/IRS-1/IRS 2 phosphorylation through upregulation of SOCS-3, which almost completely blocks Jak2 activation. PMID- 12734780 TI - Myocardial perfusion abnormalities in patients with active acromegaly. AB - Cardiomyopathy is often seen in patients with a long history of acromegaly. In order to screen for perfusion abnormalities, patients with active acromegaly without evidence for coronary heart disease were examined by single photon emission computed tomography (SPECT). The study included a group of 11 strictly selected patients with active acromegaly (7 males and 4 females; age 51 +/- 12 y [mean +/- S.D.]) with elevated age-adjusted IGF-I levels (IGF-I 569 +/- 193 micro g/l; GH 31.2 +/- 56.3 micro g/l) compared to an age- and sex-matched non acromegalic control group with comparable muscle mass index of the left ventricle (126 +/- 41 active vs. 122 +/- 33 g/m 2 control group) and body mass index (26.6 +/- 2.7 vs. 27.0 +/- 5.0 kg/m 2). To address this issue, myocardial perfusion was investigated by single photon emission computed tomography (SPECT) using a triple head gamma-camera. 70 MBq 201TlCl was injected, and post-stress (from bicycle ergometer) images were obtained. Images were interpreted quantitatively by bull's eye polary map (16 regions of the left ventricle) and were compared to the control group. In the patients with active acromegaly, the mean nuclide uptake of the 16 regions of the left ventricle after bicycle stress examination was lower than in the control group (82.99 +/- 2.85 active vs 85.48 +/- 1.29 control group, p < 0.01). Non-homogeneity of nuclide uptake was defined as the standard deviations of the 16 regions and was higher in patients with active acromegaly (11.11 +/- 2.35 active vs. 8.77 +/- 1.39 control group, p < 0.01). In conclusion, myocardial perfusion is impaired in patients with active acromegaly, thus representing an early stage of cardiac involvement in acromegaly that may be directly mediated by growth hormone excess. PMID- 12734782 TI - A molecular method for classifying the genotypes obtained in a breeding colony from testicular feminized (Tfm) rats. PMID- 12734781 TI - A one-year, randomised, multicentre trial comparing insulin glargine with NPH insulin in combination with oral agents in patients with type 2 diabetes. AB - AIMS: The aim of the trial was to compare the efficacy and safety of the new, long-acting basal insulin, insulin glargine (LANTUS(R)), with NPH human insulin, each administered in a combination regimen with oral antidiabetic drugs in patients with Type 2 diabetes. METHODS: In a multicentre, open, randomised study, 570 patients with Type 2 diabetes, aged 34 - 80 years, were treated for 52 weeks with insulin glargine or NPH insulin given once daily at bedtime. Previous oral antidiabetic therapy was continued throughout the study. RESULTS: There was a clinically relevant decrease in glycosylated haemoglobin (GHb) values from baseline to endpoint with both drugs (insulin glargine: - 0.46 %; NPH insulin: - 0.38 %; p = 0.415); also, this difference was statistically significant in the subgroup of overweight patients with BMI > 28 kg/m 2 (insulin glargine: - 0.42 %, NPH insulin: - 0.11 %; p = 0.0237). Over the entire treatment period, NPH insulin treated patients (41 %) and insulin glargine-treated patients (35 %) experienced a similar level of symptomatic hypoglycaemia. A statistically significant difference was observed in the number of patients treated with NPH insulin who reported at least one episode of nocturnal hypoglycaemia compared with those treated with insulin glargine in the overall population and in the overweight subgroup (overall: 24 % vs. 12 %, p = 0.002; overweight: 22.2 % vs. 9.5 %, p = 0.0006), using the Cochran-Mantel-Haenszel test. These differences were most pronounced in insulin-naive and overweight (BMI > 28 kg/m 2) sub-groups. The incidence of adverse events was similar for the two treatments. CONCLUSIONS: This study demonstrated that insulin glargine is as effective as NPH insulin in achieving glycaemic control in patients with Type 2 diabetes, and is associated with fewer episodes of symptomatic hypoglycaemia, particularly nocturnal episodes. PMID- 12734783 TI - Hyaluronan fragments: an information-carrying system? PMID- 12734785 TI - The effect of progesterone and synthetic progestins on serum- and estradiol stimulated proliferation of human breast cancer cells. AB - The results from the Women's Health Initiative study on enhanced breast cancer risk in postmenopausal women using an estrogen/progestin combination clearly indicate the need for a comparison of different progestins with regard to cancer risk. To shed some light on this issue, we have investigated the influence of progesterone and various synthetic C19- and C21-progestins on cell proliferation of a human breast cancer cell line in vitro. Of special interested was the comparison of two different regimens commonly used in HRT, sequential and continuous combination with estradiol. We used the human breast cancer cell line MCF-7 as a model. Progesterone (P), chlormadinone acetate (CMA), dienogest (DNG), gestodene (GSD), 3-ketodesogestrel (KDG), levonorgestrel (LNG), medroxyprogesterone acetate (MPA), and norethisterone (NET) were investigated in the range of 0.01nm to 10 micro M alone and in combination with 10 nM estradiol. Cell proliferation was measured after 7 days using the ATP chemosensitivity test. Tested alone, CMA, DNG, GSD, KDG, MPA and NET significantly stimulated cell proliferation, but only at high dosages. Sequentially combined with estradiol, only CMA inhibited cell proliferation over the whole concentration range. Slight effects were found for DNG, GSD and KDG, whereas P and MPA only showed an effect at the highest concentration. NET had no significant effect on cell proliferation. Continuously combined, all progestins exhibited an inhibitory effect over the whole concentration range. The most prominent effects were found for P, CMA, GSD, and KDG. Only slight effects were found for DNG, MPA and NET. Our in vitro results indicate that the influence on breast cancer risk using HRT in postmenopausal women might depend on the type of progestin used as well as on the regimen applied. However, the net inhibitory in vitro effect of the progestins at clinically relevant dosages is rather minimal, and whether progestins in general can reduce breast cancer risk in long-term treatment remains uncertain. Further clinical trials are urgently needed to clarify this issue. PMID- 12734784 TI - Testosterone and sexual experience alter levels of plasma membrane binding sites for progesterone in the male rat brain. AB - Physiological levels of progesterone act in conjunction with androgens to facilitate copulatory behavior in male rats, mice, and lizards. Radiolabeled progesterone conjugated to bovine serum albumin measured specific binding sites in membrane fractions from male rats that were gonadectomized and testosterone treated, or remained gonadally intact, to determine the role of gonadal steroids on mPR binding. To determine whether behavioral experience could alter binding levels, males either remained sexually naive or became sexually experienced. In sexually naive males, the highest levels of specific binding occurred in the dorsal portions of the medial preoptic area, with only moderate levels of binding in ventral portions of the medial preoptic area and the dorsal and ventral medial hypothalamus. However, conjugated progesterone binding in these brain regions did not change as a function of testosterone or behavioral manipulations. In contrast, the amygdala responded to behavioral experience with significantly (4 fold) increased binding in gonadectomized, T-treated males with sexual experience. These data indicate that the neuronal plasticity for membrane associated progesterone binding is regionally specific, being regulated by sexual experience following the reinstatement of testosterone levels, thus suggesting a functional role for plasma membrane activity of progesterone in male rat reproduction. PMID- 12734786 TI - Leptin fully suppresses acetylcholine-induced insulin secretion and is reversed by tolbutamide in isolated perfused chicken pancreas. AB - So far, there has been no evidence for any direct pancreatic effect of leptin in the chicken. The present study was aimed at detecting chicken leptin receptor (cOb-R) expression in isolated chicken islets of Langerhans and to examine the direct effect of leptin on insulin secretion after stimulation by acetylcholine (1 micro M) + glucose (14 mM) from isolated perfused chicken pancreas. We will show that i) full length cOb-R mRNA was expressed in isolated pancreatic islets of chickens, ii) recombinant chicken leptin (10 nM) or diazoxide (100 micro M) rapidly (within 2 min) and significantly suppressed insulin secretion induced by acetylcholine stimulation without any change in volume outflow rate, iii) tolbutamide (100 micro M) introduced 10 min after leptin and perfused for 10 min fully reversed the suppressive effect of leptin on pre-established acetylcholine induced insulin release. In conclusion, we found that leptin has a profound inhibitory influence upon insulin secretion in perfused chicken pancreas. The results suggest that leptin inhibits insulin secretion by acting before or at the level of K ATP channels in chicken pancreatic beta-cells. Further studies are warranted to clarify the specific inhibitory mechanism. PMID- 12734787 TI - Clinical, metabolic and endocrine parameters in response to metformin in obese women with polycystic ovary syndrome: a randomized, double-blind and placebo controlled trial. AB - There is still some controversy concerning the effects of metformin in the treatment of Polycystic Ovary Syndrome (PCOS). The aim of this study was to asses the effect of metformin on clinical, metabolic and hormone parameters in obese women with PCOS. Thirty obese, non-diabetic women with PCOS received 500 mg of metformin or placebo, TID, over 90 days. Assessed parameters included body mass index, waist-to-hip ratio, blood pressure, FSH, LH, total testosterone, SHBG, fasting insulinemia, insulin-to-glucose ratio, total, HDL and LDL cholesterol, triglycerides, and menstrual cycles before and after the use of the drugs. Before treatment, patients did not differ in the two groups. After 90 days of metformin use, PCOS women presented significantly lower levels of total testosterone (p = 0.030) and total cholesterol (p = 0.023) compared to the women that used placebo. The other parameters did not differ between the groups. In conclusion, obese women with PCOS may benefit from the use of metformin through the reduction of hyperandrogenemia, total cholesterol, and possibly by restoration of regular menstrual cycles. Further studies with longer follow-ups are necessary to determine cardiovascular and endometrial metformin benefits and insulin resistance decrease in women with polycystic ovary syndrome. PMID- 12734788 TI - Leptin levels in type 2 diabetes: associations with measures of insulin resistance and insulin secretion. AB - Interactions between leptin and insulin have been shown previously, in vitro and in vivo. In this study, we evaluate the associations of leptin levels with insulin secretion and insulin sensitivity in type 2 diabetes. Fasting leptin levels, HbA 1c, glucose, insulin, C-peptide, intact and des-31,32-proinsulin were measured in 100 non-insulin-treated type 2 diabetic patients. Glucose, insulin and C-peptide were measured 2 hours after an oral glucose load. Insulin resistance and beta-cell function were calculated using HOMA. Leptin levels were found to be associated with all measures of beta-cell secretion: with fasting and 2 hours insulin and C-peptide, with intact and des-31,32-proinsulin concentrations, and with beta-cell secretion estimated with HOMA. This association was independent of age and body fat in women, but in men, associations with insulin and C-peptide weakened after controlling for fat mass, whereas those with intact and des-31,32-proinsulin disappeared. Fasting insulin and C-peptide levels were also significant in multiple regression analyses, besides gender and fat mass. Insulin resistance, as assessed by HOMA, was strongly correlated with leptin, also after correction for age and fat mass in both genders. We conclude that, besides fat mass and gender - the main determinants for leptin levels in type 2 diabetic subjects as in healthy subjects - insulin secretion and the degree of insulin resistance also seem to contribute significantly to leptin levels. PMID- 12734789 TI - Post-exercise increase of lipid oxidation after a moderate exercise bout in untrained healthy obese men. AB - The aim of the study was to examine whether a moderate exercise increases the utilization of fatty acids during the recovery period in obese men. Six healthy obese participated in a randomized crossover investigation, one with exercise and one without exercise. At 8 a. m., the subjects had a standardized breakfast and they rested in a sitting position for 3 hours. The subjects were maintained in the sitting position for 4 additional hours in one session. In a second session, they exercised for 60 min at 50 % of their VO(2) max and then returned to the sitting position for 3 hours. Respiratory exchange ratio (RER) values were calculated by indirect calorimetry. During the resting session, plasma non esterified fatty acids (NEFA) and glycerol concentrations rose progressively, whereas RER progressively decreased. During the exercise, plasma catecholamines, NEFA, glycerol, growth hormone and cortisol levels and RER increased while insulin decreased. During the recovery, plasma NEFA increased and glycerol decreased. During the first hour of recovery, RER values were lower and fatty acid utilization higher than during the same period of the resting session. The study shows that exercise induces modifications in hormonal factors promoting lipid mobilization and suggests that exercise provide substantial amounts of NEFA for muscle oxidation during recovery from an exercise bout in obese subjects. PMID- 12734790 TI - Dehydroepiandrosterone metabolism in human plasma. AB - The possibility that dehydroepiandrosterone (DHEA) is metabolized in human plasma was studied by column and thin-layer chromatography. The results obtained indicate that a time-dependent disappearance of DHEA is matched by the appearance of newly-formed species that may represent DHEA conversion by-products. Neither disappearance of DHEA, nor formation of the alleged conversion by-products was observed when reactions were performed under conditions in which plasma enzymes were removed or inactivated. These results suggest that, in plasma, DHEA is partially transformed into different substances, and that the conversion reactions are catalyzed by enzymes present in this tissue. The observed kinetics of appearance and partial disappearance of the radiolabeled species can be interpreted as indicating that some of the by-products formed are further converted into other substances. The data shown appear to indicate that plasma can be added to the list of the already known compartments that are involved in steroid metabolism. PMID- 12734791 TI - Hormonal reactions during heavy training stress and following tapering in highly trained male rowers. AB - The purpose of this study was to determine whether fasting plasma leptin, cortisol, testosterone and growth hormone concentrations were altered with a heavy increase in training stress followed by a reduced stress in highly trained male rowers. Twelve male national standard rowers (age 20.5 +/- 3.0 years, height 187.9 +/- 6.1 cm, body mass 87.1 +/- 8.3 kg, percent body fat 10.4 +/- 3.2 %) underwent a three-week period of maximally increased training stress followed by a two-week tapering period. The fasting blood samples were obtained every week after the rest day. In addition, the maximal 2000-meter rowing ergometer performance time was assessed before and immediately after the exhaustive training period as well as after the tapering period. A 22 % increase in training stress caused a significant decrease (by 8 %) and increase (by 9 %) in leptin and testosterone, respectively. A further increase in training volume by 25 % significantly reduced leptin further by 35 %. At the same time, no changes were observed in testosterone. Growth hormone was significantly elevated only after the first week of heavy training stress compared to the pretraining level. In the first tapering week, where the physical stress was reduced by 50 %, leptin only significantly increased by 29 %. Testosterone and growth hormone were significantly reduced to almost pretraining levels by the end of the second tapering week. Leptin was further significantly increased during the second tapering week. Cortisol remained relatively constant during the whole study period. Similarly, rowing performance was not significantly changed. We conclude that leptin is more sensitive to the rapid and pronounced changes in training stress compared to measured stress hormones in athletes. In addition, fasting plasma leptin could be regarded as a key signal for metabolic adaptation to exhaustive training stress in highly trained male rowers. PMID- 12734792 TI - Impact of growth hormone on central nervous activity, vigilance, and tiredness after short-term therapy in growth hormone-deficient adults. AB - Impairment of well-being and cognitive function has been reported in growth hormone-deficient adults, as well as an improvement of these parameters after GH substitution, albeit inconsistently. The effect of growth hormone on central nervous activity, vigilance and sleepiness was studied prospectively in 16 growth hormone-deficient adults (7 females, 9 males, mean age: 36.8 yrs) with multiple pituitary hormone deficiencies before and 3 months after the start of growth hormone substitution using two objective methods of measurement, pupillographic sleepiness test and a choice reaction time test. Significant differences were found for neither pupillary unrest index nor for reaction time, false or missing reactions in 12 evaluable patients (7 females, 5 males, mean age 37.8 years). Because of the known interrelationships between growth hormone, sleep and mood, the visual analogue scale for tiredness and standardized retrospective questionnaires regarding sleep and mood (Pittsburgh sleep quality index, Epworth sleepiness scale, Depression scale) were used as additional methods. After GH substitution, there was no difference in sleep efficiency and daytime sleepiness, but some of the subjective sleep parameters (sleep quality and sleep latency) improved significantly. There was a tendency for mood improvement, too. Although results must be interpreted cautiously due to the small sample size, we conclude that the improved sleep and mood parameters might be caused by other indices of general well-being in our study. PMID- 12734793 TI - HLA-DQA1*0301-associated susceptibility for autoimmune polyglandular syndrome type II and III. AB - No significant differences were reported for the frequency of DR3-DQ2 and DR4-DQ8 haplotypes in a recent study of one of the largest cohorts worldwide of patients with isolated Addison's disease compared to patients with APS II. However, previous studies had suggested that the HLA-DQ genes, especially DQA1*0102, may be a genetic marker for resistance to autoimmune thyroid disease, which is the most frequent disease in APS II or III. Until now, HLA-DQA1 alleles have not been systematically investigated in APS. We determined the HLA-DR and HLA-DQA1 association in 112 unrelated patients with APS II (n = 29), APS III (n = 83) and 184 unrelated patients with single-component diseases without further manifestations of APS: Graves' disease (n = 70), Hashimoto's thyroiditis (n = 53), autoimmune Addison's disease (n = 15), vitiligo (n = 16) and alopecia (n = 30), and 72 healthy controls - German Caucasians - to identify possible predisposing and protective HLA class II alleles in APS. In agreement with previous studies, we detected a significantly higher frequency of DR 3 and/or DR 4 in patients with APS II and III compared to controls. In patients with APS II, we detected a significantly higher frequency of DQA1*0301 and *0501 compared to controls confirming the increased frequency of an extended HLA DRB1-*04-DQA1-*03 DQB-*03 haplotype as previously described. In contrast, only DQA1*0301 was increased in our patients with APS III compared to controls. Moreover, we detected an increased frequency of DQA1*0301 in patients with APS, whereas DQA1*0301 was only significantly elevated in alopecia in patients with single component diseases without APS. Therefore, our results indicate an association between DQA1*0301 and APS II or III since this allele was otherwise not significantly associated with any of its component diseases except alopecia. Moreover, our data imply that the allele DQA1*0301 is a marker of increased risk for further APS manifestations in patients who suffer from an organ-specific autoimmune disease. PMID- 12734794 TI - Glucose-mannose-binding lectins isolated from Brazilian beans stimulate the autophosphorylation of the insulin receptor in vitro. PMID- 12734795 TI - Occurrence, horizontal transfer and degeneration of VDE intein family in Saccharomycete yeasts. AB - VDE is a homing endonuclease gene originally discovered as an intervening element in VMA1s of Saccharomyces cerevisiae. There have been two independent subfamilies of VDE, one from S. cerevisiae strain X2180-1A and the other from Saccharomyces sp. DH1-1A in the host VMA1 gene, and they share the identity of 96.3%. In order to search the occurrence, intra/interspecies transfer and molecular degeneration of VDE, complete sequences of VMA1 in 10 strains of S. cerevisiae, eight species of saccharomycete yeasts, Candida glabrata and Kluyveromyces lactis were determined. We found that six of 10 S. cerevisiae strains contain VDEs 99.7-100% identical to that of the strain X2180-1A, one has no VDE, whereas the other three harbour VDEs 100% identical to that of the strain DH1-1A. S. carlsbergensis has two VMA1s, one being 99.8% identical to that of the strain X2180-1A with VDE 100% identical to that of the strain DH1-1A and the other containing the same VMA1 in S. pastorianus with no VDE. This and other evidence indicates that intra/interspecies transmissions of VDEs have occurred among saccharomycete yeasts. Phylogenetic analyses of VMA1 and VDE suggest that the S. cerevisiae VDEs had branched earlier than other VDEs from an ancestral VDE and had invaded into the host loci as relatively late events. The two VDEs seemed to degenerate in individual host loci, retaining their splicing capacity intact. The degeneration of the endonuclease domains was distinct and, if compared, its apparent rate was much faster than that of the protein-splicing domains. PMID- 12734796 TI - Moderately lipophilic carboxylate compounds are the selective inducers of the Saccharomyces cerevisiae Pdr12p ATP-binding cassette transporter. AB - Saccharomyces cerevisiae displays very strong induction of a single ATP-binding cassette (ABC) transporter, Pdr12p, when stressed with certain weak organic acids. This is a plasma membrane pump catalysing active efflux of the organic acid anion from the cell. Pdr12p action probably allows S. cerevisiae to maintain lower intracellular levels of several weak organic acid preservatives than would be expected on the basis of the free equilibration of the acid across the cell membrane. This in turn facilitates growth in the presence of these preservatives and therefore yeast spoilage of food materials. Pdr12p appears to confer resistance to those carboxylic acids that, to a reasonable degree, partition into both the lipid bilayer and aqueous phases. Its gene (PDR12) is strongly induced by sorbate, benzoate and certain other moderately lipophilic carboxylate compounds, but not by organic alcohols or high levels of acetate. PDR12 induction reflects the operation of a previously uncharacterized S. cerevisiae stress response, for which the induction signal is probably a high intracellular pool of the organic acid anion. PMID- 12734797 TI - A 'marker switch' approach for targeted mutagenesis of genes in Schizosaccharomyces pombe. AB - The completion of the Schizosaccharomyces pombe genome sequencing project has led to a dramatic acceleration of gene characterization in this system. Once a gene has been identified, the challenge then comes in using reverse genetics to generate a range of mutants in this gene of interest so that the powerful genetics and wealth of genetic backgrounds available in Sz. pombe can be exploited to study the function of the newly identified molecule. Beyond simple PCR-tagging approaches, the high frequency with which illegitimate recombination occurs in Sz. pombe has made the manipulation of some loci complex, time consuming and a process of trial and error. Here we describe a simple 'marker switch' approach that enables the rapid selection of integration events at the locus of interest from an excessive background of integration at heterologous sites. We use the generation of temperature-sensitive mutations in the plo1(+) gene to validate this approach. PMID- 12734798 TI - An analysis of the Candida albicans genome database for soluble secreted proteins using computer-based prediction algorithms. AB - We sought to identify all genes in the Candida albicans genome database whose deduced proteins would likely be soluble secreted proteins (the secretome). While certain C. albicans secretory proteins have been studied in detail, more data on the entire secretome is needed. One approach to rapidly predict the functions of an entire proteome is to utilize genomic database information and prediction algorithms. Thus, we used a set of prediction algorithms to computationally define a potential C. albicans secretome. We first assembled a validation set of 47 C. albicans proteins that are known to be secreted and 47 that are known not to be secreted. The presence or absence of an N-terminal signal peptide was correctly predicted by SignalP version 2.0 in 47 of 47 known secreted proteins and in 47 of 47 known non-secreted proteins. When all 6165 C. albicans ORFs from CandidaDB were analysed with SignalP, 495 ORFs were predicted to encode proteins with N-terminal signal peptides. In the set of 495 deduced proteins with N terminal signal peptides, 350 were predicted to have no transmembrane domains (or a single transmembrane domain at the extreme N-terminus) and 300 of these were predicted not to be GPI-anchored. TargetP was used to eliminate proteins with mitochondrial targeting signals, and the final computationally-predicted C. albicans secretome was estimated to consist of up to 283 ORFs. The C. albicans secretome database is available at http://info.med.yale.edu/intmed/infdis/candida/ PMID- 12734799 TI - KlROM2 encodes an essential GEF homologue in Kluyveromyces lactis. AB - Cellular integrity in yeasts is ensured by a rigid cell wall whose synthesis is controlled by a MAP kinase signal transduction cascade. In Saccharomyces cerevisiae upstream regulatory components of this MAP kinase pathway involve a single protein kinase C, which is regulated in part by interaction with the small GTPase Rho1p. This small G protein is in turn rendered inactive (GDP-bound) or is activated (GTP-bound) by the influence of GTPase activating proteins (GAPs) and the GDP/GTP exchange factors (GEFs), respectively. We report here on the isolation of a gene from Kluyveromyces lactis, KlROM2, which encodes a member of the latter protein family. The nucleotide sequence contains an open reading frame of 1227 amino acids, with an overall identity of 57% to the Rom2 protein of S. cerevisiae. Four conserved sequence motifs could be identified: a RhoGEF domain, a DEP sequence, a CNH domain and a less conserved pleckstrin homology (PH) sequence. Klrom2 null mutants show a lethal phenotype, which indicates that the gene may encode the only functional GEF regulating the cellular integrity pathway in K. lactis. Conditional genomic expression of KlROM2 resulted in sensitivity towards caffeine and Calcofluor white as typical phenotypes of mutants defective in this pathway. Overexpression of KIROM2 from multicopy plasmids under the control of the ScGAL1 promoter severely impaired growth in both S. cerevisiae and in K. lactis. The fact that the lethal phenotype was not prevented in mpk1 deletion mutants indicates that growth inhibition is not simply caused by hyperactivation of the Pkc1p signal transduction pathway. PMID- 12734800 TI - Guanidine reduces stop codon read-through caused by missense mutations in SUP35 or SUP45. AB - Sup35 and Sup45 are essential protein components of the Saccharomyces cerevisiae translation termination factor. Yeast cells harbouring the [PSI(+)] prion form of Sup35 have impaired stop codon recognition (nonsense suppression). It has long been known that the [PSI(+)] prion is not stably transmitted to daughter cells when yeast are grown in the presence of mM concentrations of guanidine hydrochloride (GuHCl). In this paper, Mendelian suppressor mutations whose phenotypes are likewise hidden during growth in the presence of millimolar GuHCl are described. Such GuHCl-remedial Mendelian suppressors were selected under conditions where [PSI(+)] appearance was limiting, and were caused by missense mutations in SUP35 or SUP45. Clearly, anti-suppression caused by growth in the presence of GuHCl is not sufficient to distinguish missense mutations in SUP35 or SUP45, from [PSI(+)]. However, the Mendelian and prion suppressors can be distinguished by subsequent growth in the absence of GuHCl, where only the nonsense suppression caused by the [PSI(+)] prion remains cured. Recent reports indicate that GuHCl blocks the inheritance of [PSI(+)] by directly inhibiting the activity of the protein remodelling factor Hsp104, which is required for the transmission of [PSI(+)] from mother to daughter cells. However, the nonsense suppressor activity caused by the GuHCl-remedial sup35 or sup45 suppressors does not require Hsp104. Thus, GuHCl must anti-suppress the sup35 and sup45 mutations via an in vivo target distinct from Hsp104. PMID- 12734801 TI - Isolation of the MNN9 gene of Yarrowia lipolytica (YlMNN9) and phenotype analysis of a mutant ylmnn9 Delta strain. AB - In this work we describe the isolation of the Yarrowia lipolytica homologue of Saccharomyces cerevisiae MNN9 gene, which we have named YlMNN9, and the phenotype analysis of a Y. lipolytica strain containing the disrupted YlMNN9 allele. YlMNN9 was cloned using degenerate consensus oligonucleotides to generate specific probes that were in turn used to screen mini-gene libraries. The gene is defined by a 1014 bp ORF predicted to encode a protein 337 amino acids long that shares significant homology with the Mnn9ps of S. cerevisiae, Candida albicans and Hansenula polymorpha, including a putative N-terminal transmembrane domain. Disruption of YlMNN9 leads to phenotypes such as resistance to sodium orthovanadate and sensitivity to hygromycin B, compatible with a glycosylation defect, and hypersensitivity to Calcofluor white, Congo red or zymolyase, characteristic of cell wall defects. Analysis of cell wall proteins present in beta-mercaptoethanol and zymolyase extracts showed significant differences between the parental and the ylmnn9 Delta strain. These results suggest that, as has been the case with the mnn9 strain of S. cerevisiae, the ylmnn9 Delta strain we present in this work, could be used to study the cell wall proteins of Y. lipolytica and how they are organized into the cell wall. PMID- 12734802 TI - Gene regulation in response to protein disulphide isomerase deficiency. AB - We have examined the activities of promoters of a number of yeast genes encoding resident endoplasmic reticulum proteins, and found increased expression in a strain with severe protein disulphide isomerase deficiency. Serial deletion in the promoter of the MPD1 gene, which encodes a PDI1-homologue, revealed a cis acting element responding to deficiency of protein disulphide isomerase activity (designated CERP). The presence of the sequence element is necessary and sufficient for the upregulation in response to disulphide isomerase deficiency, as measured by a minimal promoter containing the CERP element. The sequence (GACACG) does not resemble the unfolded protein response element. It is present in the upstream regions of the MPD1, MPD2, KAR2, PDI1 and ERO1 genes. PMID- 12734803 TI - Perspectives on blood/materials interactions. PMID- 12734804 TI - Structural degradation of acrylic bone cements due to in vivo and simulated aging. AB - Acrylic bone cement is the primary load-bearing material used for the attachment of orthopedic devices to adjoining bone. Degradation of acrylic-based cements in vivo results in a loss of structural integrity of the bone-cement-prosthesis interface and limits the longevity of cemented orthopedic implants. The purpose of this study is to investigate the effect of in vivo aging on the structure of the acrylic bone cement and to develop an in vitro artificial aging protocol that mimics the observed degradation. Three sets of retrievals are examined in this study: Palacos brand cement retrieved from hip replacements, and Simplex brand cement retrieved from both hip and knee replacement surgeries. In vitro aging is performed using oxidative and acidic environments on three acrylic-based cements: Palacos, Simplex, and CORE. Gel permeation chromatography (GPC) and Fourier transform infrared spectroscopy (FTIR) are used to examine the evolution of molecular weight and chemical species within the acrylic cements due to both in vivo and simulated aging. GPC analysis indicates that molecular weight is degraded in the hip retrievals but not in the knee retrievals. Artificial aging in an oxidative environment best reproduces this degradation mechanism. FTIR analysis indicates that there exists a chemical evolution within the cement due to in vivo and in vitro aging. These findings are consistent with scission-based degradation schemes in the cement. Based on the results of this study, a pathway for structural degradation of acrylic bone cement is proposed. The findings from this investigation have broad applicability to acrylic-based cements and may provide guidance for the development of new bone cements that resist degradation in the body. PMID- 12734805 TI - Visualization of specific interaction between biomimetic glycopolymer containing reducing glucose moiety and HepG2 cell mediated by GLUT-1. AB - Poly [3-O-(4'-vinylbenzyl)-D-glucose] (PVG) is a polystyrene derivative that contains glucose moieties, which interact with glucose transporter (GLUT-1) carrying HepG2 cells. To clarify the specific interaction between the PVG and HepG2 cells, PVG polymer labeled with fluorescent rhodamine-B isothiocyanate (RITC) was used to follow the specific interaction, which was visualized by confocal laser microscopy. We found that PVG binds strongly to HepG2 cells, probably because of a specific interaction mediated by the presence of GLUT-1 on the cell membrane. The fluorescence intensity of PVG and HepG2 cells was up to fourfold (0.11 +/- 0.04) that of any other glycopolymer and HepG2 cell interaction (0.025 +/- 0.01). Moreover, cellular fluorescence increased significantly on increasing the incubation time and the polymer concentration. To verify the specific nature of the interaction between PVG and HepG2 cells, the cells were pretreated with phloretin, an inhibitor of GLUT-1 before adding RITC labeled PVG polymer to the cell culture medium; this treatment was found to suppress the action of PVG on HepG2 cells. PMID- 12734806 TI - Morphological studies on the culture of kidney epithelial cells in a fiber-in fiber bioreactor design with hollow fiber membranes. AB - A hollow fiber-in-fiber-based bioreactor system was tested for the applicability to host kidney epithelial cells as a model system for a bioartificial kidney. Hollow fibers were prepared from polyacrylonitrile (PAN), polysulfone polyvinylpyrollidinone (PVP) blend (PSU) and poly(acrylonitrile-N vinylpyrollidinone) copolymer P(AN-NVP). Hollow fibers with smaller and larger diameters were prepared so that the smaller fitted into the larger, with a distance of 50-100 microm in between. The following material combinations as outer and inner fiber were applied: PAN-PAN; PSU-PSU, PSU-P(AN-NVP). Madin-Darby kidney epithelial cells (MDCK) were seeded in the interfiber space and cultured for a period up to 14 days. Light, scanning, and transmission electron microscopy were used to follow the adhesion and growth of cells, and to characterize their morphology. As a result, we found that MDCK cells were able to grow in the interfiber space in mono- and multilayers without signs of systemic degeneration. Comparison of the different materials showed that PAN and P(AN-NVP) provided the best growth conditions, indicated by a tight attachment of cells on hollow fiber membrane, and subsequent proliferation and development of structural elements of normal epithelia, such as tight junctions and microvilli. In conclusion, the fiber-in-fiber design seems to be an interesting system for the construction of a bioartificial kidney. PMID- 12734807 TI - Formation of hydroxyapatite in water, Hank's solution, and serum at physiological temperature. AB - The influence of de-ionized water, Hank's saline solution, and bovine calf serum on formation of stoichiometric (Ca/P = 1.67) hydroxyapatite (SHAp) at physiological temperature was studied. SHAp formed in aqueous solution by acid base reaction of particulate Ca(H(2)PO(4))(2).H(2)O and Ca(4)(PO(4))(2)O. Hydroxyapatite formation is accompanied by an initial period of surface hydration of the precursors, an induction period, and a period during which the bulk of the conversion to hydroxyapatites occurs. The formation of SHAp occurred more rapidly in Hank's solution and distilled water than in serum. The formation of SHAp from these precursors is strongly inhibited by serum. There were two primary exothermal events associated with SHAp formation: initial heat evolution peak, which was associated with reactant dissolution, and the major heat evolution peak, which was associated with SHAp formation. The presence of the constitutents in serum depresses both. This is a result of serum macromolecules adsorbing onto the available surfaces regardless of whether they are reactants or products. Variations in heat evolution behavior, pH, and the times of disappearance of the reactants and appearance of SHAp correlate with one another. PMID- 12734808 TI - Reduced adhesion of blood cells to biodegradable polymers by introducing phosphorylcholine moieties. AB - Aliphatic polyesters are believed to be good biocompatible polymers for tissue engineering because of their biodegradability and nontoxicity of the degradated products. However, it is necessary to reduce the nonspecific protein adsorption for the application of biodegradable polymers to drug delivery systems or antiadhesive membranes. We hypothesized that novel biodegradable polymers could be synthesized by introducing phosphorylcholine moieties into aliphatic polyesters. The L-lactide was polymerized in the presence of L-alpha glycelophosphorylcholine (LGPC) using stannous octate as the catalyst. The molecular weight and crystallinity of poly(L-lactide) (PLLA)-based phospholipid polymers (PLLA-PC) decreased with an increase in the composition of the LGPC unit in the PLLA-PC. The hydrolysis of the PLLA-PC was evaluated by soaking the polymer membranes in a phosphate buffer solution. The rate of weight loss was increased with increasing the LGPC units in PLLA-PC. The surface analysis of the membranes using an X-ray photoelectron microscope showed the composition of phosphorylcoline groups on the surface. The amount of adsorbed protein and adherent blood cell on the polymer surface was decreased with introducing LGPC unit. PLLA-PC is a promising biodegradable polymer having blood compatibility and antiadhesive property. PMID- 12734809 TI - Coaxial double-tubular compliant arterial graft prosthesis: time-dependent morphogenesis and compliance changes after implantation. AB - In order to reduce the compliance mismatch between the native artery and the artificial graft, we have developed a coaxial double-tubular compliant graft, using multiply micropored segmented polyurethane (SPU) thin films, which mimics the relationship between the intraluminal pressure and vessel internal diameter (P-D) of the native artery (termed "J" curve). The graft was coaxially assembled by inserting a high-compliance inner tube with a heparin-immobilized photocured gelatin coating layer into a low-compliance outer tube with a photocured hydrophilic polymer coating layer. Twenty-eight coaxial double-tubular compliant grafts were implanted into the canine common carotid arteries in an end-to-end fashion for up to 12 months. The overall patency rate was 86% (24/28), and neither rupture nor aneurysmal formation was observed. A neoarterial wall was formed via transanastomotic and transmural tissue ingrowth, resulting in neoarterial tissue formation on the luminal surface and into the intertubular space of the double-tubular graft, accompanied by mainly myofibroblasts and inflammatory cells in the early stage and endothelialization and collagen-rich extracellular matrices in the late stage of implantation. Surrounding-tissue adhesion with the outer tube was prevented by the hydrophilic polymer coating. Although the J curve of the implanted prototype model was preserved 1 month after implantation, the impaired J curves were observed because of tissue ingrowth and tissue adhesion between the outer surface of the inner tube and the surrounding tissues 3 and 6 months after implantation. At 12 months after implantation, however, the implanted coaxial double-tubular graft exhibited high compliance due to biodegradation of the SPU films. PMID- 12734810 TI - Hydrogen embrittlement of Ni-Ti superelastic alloy in fluoride solution. AB - Hydrogen embrittlement of Ni-Ti superelastic alloy in a fluoride solution (0.2% APF) has been investigated by means of a tensile test (after immersion) and hydrogen thermal desorption analysis. Upon immersion, the tensile strength of the alloy decreased to the critical stress level of martensite transformation. Hydrogen desorption of the immersed specimens appeared with a peak at around 500 degrees C. The amount of absorbed hydrogen in the alloy ranged from 100 to 1000 mass ppm when immersed in the fluoride solution for 2 to 24 h. The immersion in the fluoride solution led to the degradation of mechanical properties due to hydrogen embrittlement. The results of the present study imply that one reason that Ti and its alloys fracture in the oral cavity is the fact that hydrogen is absorbed in a fluoride solution, such as prophylactic agents. PMID- 12734811 TI - Preparation and assessment of revised simulated body fluids. AB - A simulated body fluid (SBF) with ion concentrations approximately equal to those of human blood plasma has been used widely for in vitro assessment of the bioactivity of artificial materials and for the formation of bone-like apatite on various substrates. The ion concentrations of a conventional SBF (c-SBF) are, however, not exactly equal to those of blood plasma. In the present study, a revision of c-SBF was made to prepare new SBFs (r-SBF, i-SBF, and m-SBF) with ion concentrations equal to or closer to those of blood plasma. The ion concentrations of the r-SBF and i-SBF were designed to be equal to those of blood plasma in total and dissociated amounts, respectively. The m-SBF was designed to have a total ion concentration equal to that of blood plasma, except for the concentration of HCO(-) (3), which was set to the saturated level with respect to calcite. The ion concentrations and pH of the as-prepared new SBFs were found to be equal to those of the nominal values. Upon sealed storage, the r-SBF and i-SBF showed no change in ion concentrations for up to 4 weeks at 5 degrees C, and up to 2 weeks at 36.5 degrees C, but thereafter they showed a decrease in HCO(-) (3) concentration and an increase in pH. Under the same storage conditions, the c-SBF and m-SBF showed no change in ion concentrations and pH values over a period of up to 8 weeks. These results indicate that the r-SBF and i-SBF are less stable than the c-SBF and m-SBF in terms of changes in ion concentrations relative to storage period. The m-SBF is optimal for in vitro bioactivity assessment of artificial materials and for biomimetic production of bone-like apatite. PMID- 12734812 TI - Injectable drug-delivery systems based on supramolecular hydrogels formed by poly(ethylene oxide)s and alpha-cyclodextrin. AB - Polymeric hydrogels long have attracted interest for biomaterials applications because of their generally favorable biocompatibility. High in water content, they are particularly attractive for delivery of delicate bioactive agents, such as proteins. However, because they require covalent crosslinking for gelation, many hydrogels can be applied only as implantables, and incorporation of drugs by sorption may be time-consuming and limiting with regard to the loading level. Therefore a delivery formulation where gelation and drug loading can be achieved simultaneously, taking place in an aqueous environment and without covalent crosslinking, would be attractive. Herein is described a new class of injectable and bioabsorbable supramolecular hydrogels formed from poly(ethylene oxide)s (PEOs) and alpha-cyclodextrin (alpha-CD) for controlled drug delivery. The hydrogel formation is based on physical crosslinking induced by supramolecular self-assembling with no chemical crosslinking reagent involved. The supramolecular structure of the hydrogels was confirmed with wide-angle X-ray diffraction studies. The gelation kinetics was found to be dependent on the concentrations of the polymer and alpha-CD as well as on the molecular weight of the PEO used. The rheologic studies of the hydrogels showed that the hydrogels were thixotropic and reversible and that they could be injected through fine needles. The components of the supramolecular hydrogels potentially are biocompatible and nontoxic. Drugs can be encapsulated directly into the hydrogels in situ at room temperature without any contact with organic solvents. The supramolecular hydrogels were evaluated in terms of their in vitro release kinetics. The rate-controlling mechanism of macromolecular drug release might be the erosion of the hydrogels. PMID- 12734813 TI - Antibacterial characteristics of newly developed amphiphilic lipids and DNA-lipid complexes against bacteria. AB - The purpose of this study was to investigate the antibacterial activity of newly developed amphiphilic lipids and DNA/lipid complexes against two types of oral bacteria and two types of hospital infection bacteria. Nine amphiphilic lipids were quantitatively prepared from the reaction of n-alkyl alcohol, alpha-amino acids, and p-toluenesulfonic acid. Nine DNA-lipid complexes were prepared by the simple mixing of DNA and amphiphilic lipids. The DNA-lipid complexes were insoluble in water. The antibacterial activity of lipids and DNA-lipid complexes against Porphyromonas gingivalis, Streptococcus mutans, Staphylococcus aureus, and Pseudomonas aeruginosa were evaluated by the disk-diffusion method. Seven artificial lipids showed antibacterial behavior; in particular, the lipids prepared from n-decyl alcohol and glycine and from n-decyl alcohol and L-alanine showed antibacterial activity against the four bacterial strains used in this study. On the other hand, the lipids of glutamic acid derivatives did not show any antibacterial activity against the four bacteria strains except for the lipid with an n-octyl group. Five DNA-lipid complexes also had an antibacterial effect. The complex prepared from DNA and glycine decyl ester p-toluenesulfonic acid salt exhibited antibacterial activity against the four types of bacteria strains. In this study it was found that lipids and DNA-lipid complexes with a mono-decyl group or a mono-dodecyl group have more favorable antibacterial activity. PMID- 12734814 TI - Enhanced solubility of paclitaxel using water-soluble and biocompatible 2 methacryloyloxyethyl phosphorylcholine polymers. AB - The purpose of this study was to enhance the water-solubility of paclitaxel (PTX) using an amphiphilic 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer as the solubilizer. PTX is an antineoplastic drug effective for various cancers, especially for ovarian and breast cancers. However, its solubility in aqueous medium is quite low, less than 0.1 microg/mL in water. We prepared the amphiphilic MPC polymers containing hydrophobic units to form aggregates and provide hydrophobic domains in water. The most effective polymer to dissolve the PTX was poly[MPC-co-n-butyl methacrylate(BMA) (PMB30W)] with 70 mol % of the BMA unit. The inside polarity of PMB30W aggregate was the same as that of ethanol, which is a good solvent for dissolving PTX. The diameter of PMB30W aggregate containing 1 mg/mL of PTX was 50 nm in aqueous medium. The concentration of PTX in the PMB30W aqueous solution reached 5.0 mg/mL. The solution was transparent and PTX did not precipitate even when the solution was stored at room temperature for 1 month. Animal experiments indicated that the PMB30W has no adverse effect even when the polymer solution is injected into the bloodstream. From the PMB30W/PTX solution, we prepared by the solvent evaporation method a PMB30W film containing PTX with good transparency. The PMB30W film containing PTX easily dissolved in water to give a clear solution. We conclude that the water-soluble amphiphilic MPC polymers are good solubilizers for PTX as injectable and biocompatible drug formulations. PMID- 12734815 TI - Influence of surfactant molecules as air-entraining agent for bone cement macroporosity. AB - Calcium phosphate bone cements (CPBCs) represent a potential synthetic alternative to bone-graft materials in bone surgery applications. CPBCs are biocompatible, bioresorbable, and slowly are replaced by new bone in vivo. However, CPBCs do not develop a macroporosity during setting that would allow fast bone ingrowth and good osteointegration of the implant. For this reason, recent research has approached the problem of inducing macroporosity inside the bone cement without influencing its normal setting. In this study, a new method for obtaining injectable macroporous CPBCs is proposed. It is based on the use of sodium dodecyl sulphate (SDS) as an air-entraining agent. The results have shown that the liquid-to-powder ratio and the SDS concentration, as well as the diameter and the interconnectivity of the macropores, can control the micro- and macroporosity. This new technology can be used to develop and optimize new commercial products for osteoporotic bone filling applications. Furthermore, the presented method also can be used at low temperatures before an operation to produce preformed implants to fit the particular needs of a patient. PMID- 12734816 TI - Preparation, characterization, and in vitro release of gentamicin from coralline hydroxyapatite-alginate composite microspheres. AB - In this work, composite microspheres were prepared from bioactive ceramics such as coralline hydroxyapatite [Ca(10)(PO(4))(6)(OH)(2)] granules, a biodegradable polymer, sodium alginate, and an antibiotic, gentamicin. Previously, we have shown a gentamicin release from coralline hydroxyapatite granules-chitosan composite microspheres. In the present investigation, we attempted to prepare composite microspheres containing coralline hydroxyapatite granules and sodium alginate by the dispersion polymerization technique with gentamicin incorporated by absorption method. The crystal structure of the composite microspheres was analyzed using X-ray powder diffractometer. Fourier transform infrared spectra clearly indicated the presence of per-acid of sodium alginate, phosphate, and hydroxyl groups in the composite microspheres. Scanning electron micrographs and optical micrographs showed that the composite microspheres were spherical in shape and porous in nature. The particle size of composite microspheres was analyzed, and the average size was found to be 15 microns. The thermal behavior of composite microspheres was studied using thermogravimetric analysis and differential scanning calorimetric analysis. The cumulative in vitro release profile of gentamicin from composite microspheres showed near zero order patterns. PMID- 12734817 TI - Study of the toughening mechanisms in bone and biomimetic hydroxyapatite materials using Raman microprobe spectroscopy. AB - A Raman microprobe spectroscopy characterization of microscopic fracture mechanisms is presented for a natural hydroxyapatite material (cortical bovine femur) and two synthetic hydroxyapatite-based materials with biomimetic structures-a hydroxyapatite skeleton interpenetrated with a metallic (silver) or a polymeric (nylon-6) phase. In both the natural and synthetic materials, a conspicuous amount of toughening arose from a microscopic crack-bridging mechanism operated by elasto-plastic stretching of unbroken second-phase ligaments along the crack wake. This mechanism led to a rising R-curve behavior. An additional micromechanism, responsible for stress relaxation at the crack tip, was recognized in the natural bone material and was partly mimicked in the hydroxyapatite/silver composite. This crack-tip mechanism conspicuously enhanced the cortical bone material resistance to fracture initiation. A piezo spectroscopic technique, based on a microprobe measurement of 980 cm(-1) Raman line of hydroxyapatite, enabled us to quantitatively assess in situ the microscopic stress fields developed during fracture both at the crack tip and along the crack wake. Using the Raman piezo-spectroscopy technique, toughening mechanisms were assessed quantitatively and rationally related to the macroscopic fracture characteristics of hydroxyapatite-based materials. PMID- 12734818 TI - Investigation of elasto-mechanical properties of alginate microcapsules by scanning acoustic microscopy. AB - High-frequency scanning acoustic microscopy (SAM) was used for investigation of acoustic impedance and 3D-surface topography of full alginate microspheres that act as model of artificial biological cells. Elasto-mechanical properties of the investigated specimens have been characterized by acoustic impedance. Mean surface impedance of microspheres (diameter: 300 microm) was measured with SAM at 900 MHz with a spatial resolution of 1.5 microm. The sensitivity and reproducibility of SAM had to be increased considerable to receive and quantify signals in the very low impedance region. The multilayer analysis method was used to get quantitative data of acoustic impedance with SAM at a microscopic level. 3D images show details of structure and surface topography. As a reference, bulk measurements were performed on full alginate cylinders. The acoustical impedance and the mechanical stiffness c(11) were obtained from mass density and longitudinal ultrasound velocity at 6 MHz. The impedances received with both methods are in close agreement. The results demonstrate the SAM as a powerful tool for characterizing mechano-elastical parameters as well as surface structure and topography of microspheres with high spatial resolution. PMID- 12734819 TI - Fiber-enriched double-setting calcium phosphate bone cement. AB - Calcium phosphate bone cements are useful in orthopedics and traumatology, their main advantages being their biocompatibility and bioactivity, which render bone tissue osteoconductive, providing in situ hardening and easy handling. However, their low mechanical strength, which, in the best of cases, is equal to the trabecular bone, and their very low toughness are disadvantages. Calcium phosphate cement compositions with mechanical properties more closely resembling those of human bone would broaden the range of applications, which is currently limited to sites subjected to low loads. This study investigated the influence of added polypropylene, nylon, and carbon fibers on the mechanical properties of double setting alpha-tricalcium phosphate-based cement, using calcium phosphate cement added to an in situ polymerizable acrylamide-based system recently developed by the authors. Although the addition of fibers was found to reduce the compression strength of the double-setting calcium phosphate cement because of increased porosity, it strongly increased the cement's toughness (J(IC)) and tensile strength. The composites developed in this work, therefore, have a potential application in shapes subjected to flexure. PMID- 12734820 TI - Loss of chondroitin 6-sulfate and hyaluronan from failed porcine bioprosthetic valves. AB - Explanted porcine bioprosthetic valves have a thinned spongiosa, partially because of an overall loss of glycosaminoglycans (GAGs). We measured the concentrations of specific GAG classes in explanted bioprosthetic valves (n = 14, implanted 12.0 +/- 4.7 years) compared with glutaraldehyde-fixed porcine controls. After extraction with NaOH, GAGs were analyzed using either a hexuronic acid assay or fluorophore-assisted carbohydrate electrophoresis to quantify the individual GAG classes. The total GAG concentration in explants was 198 +/- 95 pmol/mg wet weight-93% less than freshly fixed controls. Explants also contained altered proportions of the different GAG classes relative to controls. The proportions of hyaluronan and chondroitin/dermatan-6-sulfate were reduced from 39 to 7% and 34 to 18% of total GAGs, respectively. The predominant explant GAG class was chondroitin/dermatan-4-sulfate (proportion elevated from 14 to 70%). This GAG is commonly found in the collagen-associated proteoglycan decorin, which is likely well crosslinked by glutaraldehyde. Chondroitin-6-sulfate is commonly found in the water- and hyaluronan-binding proteoglycan versican, which is likely poorly crosslinked. The loss of versican and its associated water-binding capacity is consistent with the thinned spongiosa. The resultant compromise of hydration, compressive resistance, and viscoelasticity may be responsible for the deterioration of the bioprosthesis in vivo. PMID- 12734821 TI - Adhesion and migration of marrow-derived osteoblasts on injectable in situ crosslinkable poly(propylene fumarate-co-ethylene glycol)-based hydrogels with a covalently linked RGDS peptide. AB - Marrow-derived osteoblasts were cultured on poly(propylene fumarate-co-ethylene glycol) (P(PF-co-EG)) based hydrogels modified in bulk with a covalently linked RGDS model peptide. A poly(ethylene glycol) spacer arm was utilized to covalently link the peptide to the hydrogel. Three P(PF-co-EG) block copolymers were synthesized with varying poly(ethylene glycol) block lengths relative to poly(ethylene glycol) spacer arm. A poly(ethylene glycol) block length of nominal molecular weight 2000 and spacer arm of nominal molecular weight 3400 were found to reduce nonspecific cell adhesion and show RGDS concentration dependent marrow derived osteoblast adhesion. A concentration of 100 nmol/mL RGDS was sufficient to promote adhesion of 84 +/- 17% of the initial seeded marrow-derived osteoblasts compared with 9 +/- 1% for the unmodified hydrogel after 12 h. Cell spreading was quantified as a method for evaluating adhesivity of cells to the hydrogel. A megacolony migration assay was utilized to assess the migration characteristics of the marrow-derived osteoblasts on RGDS modified hydrogels. Marrow-stromal osteoblasts migration was greater on hydrogels modified with 100 nmol/mL linked RGDS when compared with hydrogels modified with 1000 nmol/mL linked RGDS, while proliferation was not affected. These P(PF-co-EG) hydrogels modified in the bulk with RGDS peptide are potential candidates as in situ forming scaffolds for bone tissue engineering applications. PMID- 12734822 TI - Genipin-crosslinked gelatin microspheres as a drug carrier for intramuscular administration: in vitro and in vivo studies. AB - Gelatin microspheres have been widely evaluated as a drug carrier. Nevertheless, gelatin dissolves rather rapidly in aqueous environments, making the use of the polymer difficult for the production of long-term delivery systems. This adverse aspect requires the use of a crosslinking agent in forming nonsoluble networks in microspheres. However, the use of crosslinking agents such as formaldehyde and glutaraldehyde can lead to toxic side effects owing to residual crosslinkers. In an attempt to overcome this problem, a naturally occurring crosslinking agent (genipin) was used to crosslink gelatin microspheres as a biodegradable drug delivery system for intramuscular administration. Glutaraldehyde was used as a control. In the in vitro study, the morphology, dynamic swelling, and antienzymatic degradation of test microspheres were evaluated. In the in vivo study, the biocompatibility and degradability of test microspheres were implanted in the skeletal muscle of a rat model via intramuscular injection. The results obtained in the study suggested that crosslinking of gelatin microspheres with glutaraldehyde or genipin may produce distinct crosslinking structures. The water transport mechanism in both the glutaraldehyde- and genipin-crosslinked gelatin microspheres exhibit anomalous behavior ranging from Fickian to Case-II extremes. The increase of the swelling diameter for the genipin-crosslinked microspheres was significantly less than that observed for the glutaraldehyde-crosslinked microspheres. In the animal study, it was found that the degree in inflammatory reaction for tissues implanted with the genipin-crosslinked microspheres was significantly less than that implanted with the glutaraldehyde-crosslinked microspheres. Additionally, the degradation rate of the genipin-crosslinked microspheres was significantly slower than their glutaraldehyde-crosslinked counterparts. These results indicated that the genipin-crosslinked gelatin microspheres may be used as a long-acting drug carrier for intramuscular administration. PMID- 12734823 TI - Deposition of bone-like apatite on silk fiber in a solution that mimics extracellular fluid. AB - The fabrication of apatite-organic polymer hybrids is one of several attractive methods for the development of biomaterials as a substitute for bone. Such materials have both bone-bonding ability and mechanical properties analogous to natural bone. The biomimetic process has focused attention on fabricating such hybrids, where bone-like apatite is deposited on an organic polymer surface in solutions that mimic physiological conditions. In this process, a bone-like apatite layer can be coated onto organic substrates either by using a simulated body fluid (SBF) with ion concentrations nearly equal to those of human extracellular fluid, or by using fluids that are supersaturated with respect to apatite at ambient conditions. In this study, we investigated the ability of natural silk and its related materials to facilitate apatite deposition under biomimetic conditions. Cloths made of raw silk or normal silk fibers were soaked in 1.5SBF, which has 1.5 times the ion concentration of SBF. Sericin film, which is made from an extract of degummed raw silk, was soaked in 1.5SBF. The cloth and the film soaked in 1.5SBF then were characterized by scanning electron microscopic (SEM) observation, energy dispersive X-ray microanalysis (EDX), and thin-film X-ray diffraction (TF-XRD). Apatite deposition was observed on the surface of cloth made from raw silk fiber after it was soaked in 1.5SBF, but it was not observed on cloth made from normal silk fibers. The apatite deposition on the raw silk fiber cloth was accelerated when the fibers were subjected to treatment with CaCl(2) solution at a concentration of at least 1 kmol/m(3) before immersion in 1.5SBF. Apatite deposition also was observed on the sericin film after the film was soaked in 1.5SBF for 7 days. These results indicate that apatite deposition on raw silk cloth is attributable to the catalytic effect of sericin because the surface of raw silk consists of sericin whereas that of normal silk contains fibroin. The deposition of the apatite and its crystal growth are accelerated by the presence of calcium ions on the sericin after treatment with CaCl(2) solution. Thus, sericin on natural silk fiber has the potential to facilitate apatite deposition and can be useful as a polymer material in the fabrication of hybrid materials analogous to bone through biomimetic processes. PMID- 12734824 TI - Human serum opsonization of orthopedic biomaterial particles: protein-binding and monocyte/macrophage activation in vitro. AB - Wear particles generated after total joint arthroplasty activate monocyte/macrophages and incite formation of a granulomatous periprosthetic tissue associated with bone loss and implant loosening. This study tested the hypothesis that selective opsonization of orthopedic implant biomaterial wear particles by human serum proteins influences monocyte/macrophage activation. Serum protein binding to metallic, polymeric, and ceramic particles was determined by one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Individual proteins bound to particles were subsequently identified using two-dimensional SDS-PAGE, microsequencing techniques, and SWISS-PROT analysis. Effects of selective protein opsonization on particle-induced monocyte/macrophage activation were assessed by quantification of interleukin-1beta, interleukin-6, and tumor necrosis factor-alpha release. Results from one-dimensional gel analyses revealed distinct serum protein-binding patterns specific for each material tested. Two-dimensional gel analysis together with amino acid sequencing of the prominent protein species confirmed the presence of albumin and alpha-1-antitrypsin bound to all particles tested. In contrast to the metallic particles, apolipoprotein was a major species associated with polymeric particles. Opsonization of PMMA particles with purified preparations of each of the identified proteins showed that albumin significantly enhanced particle-induced monocyte/macrophage activation. These data confirm orthopedic biomaterial specific binding of human serum proteins and demonstrate that albumin exacerbates particle-induced monocyte/macrophage activation. Alterations in the chemical and surface properties of orthopedic biomaterials to modulate protein interactions may improve implant longevity. PMID- 12734825 TI - Dissolution properties of calcium phosphate granules with different compositions in simulated body fluid. AB - Calcium phosphate granules were produced through the calcination of three hydroxyapatites (HAs) at 1150 degrees C: nearly-stoichiometric (NS-HA), calcium deficient (CD-HA), and carbonated (Carb-HA). The characterization using scanning electron microscope (SEM), X-ray diffraction (XRD), and Fourier-transformed infrared spectroscopy (FT-IR) showed that those granules exhibit strong differences in chemical composition, surface texture, and dissolution behavior. Sample dissolution in a simulated body fluid (SBF) and precipitation of a calcium phosphate layer on the granule surface were followed up for 7 days by chemical analysis of calcium content in SBF. After 80-min reaction, precipitation of new calcium phosphate phases predominates over the dissolution of original granules. Comparison between SEM images and XRD/FT-IR analysis revealed that the composition of the calcium phosphate layer and its precipitation rate depend on the HA composition and its heat treatment. Calcined carbonated apatite induces the precipitation of an octacalcium phosphate (OPC) layer, whereas a carbonated apatite layer is formed from calcined-deficient HA. The calcined nearly stoichiometric HA exhibits low efficiency in producing carbonated apatite layer. PMID- 12734826 TI - Preparation and culture of hepatocyte on gelatin microcarriers. AB - Porous gelatin microcarriers having a diameter of 80-100 microm were prepared by the suspension method using toluene as the oil phase. Rat hepatocytes were cultured on gelatin and cytodexIII microcarriers. The cells retained its spherical shape, which is similar in vivo, and showed no morphological changes to the flat state. Hepatocyte aggregates on microcarriers maintained higher metabolic functions than monolayer cells. Pore size of microcarrier plays an important role in the attachment and metabolic function of cells in culture. Phase-contrast micrograph and cell activity showed that hepatocytes cultures on gelatin microcarrier of <1 microm pore size is better than that of 5-10 microm. PMID- 12734827 TI - Abstracts of the Association of Surgeons of Great Britain and Ireland Annual Meeting. Manchester, United Kingdom, 7-9 May 2003. PMID- 12734853 TI - 'Lean' surgery through changes in surgical work flow. PMID- 12734854 TI - Screening for abdominal aortic aneurysm. PMID- 12734855 TI - Ovarian ablation in the adjuvant treatment of premenopausal and perimenopausal breast cancer. AB - BACKGROUND: Ovarian ablation has been employed in the treatment of breast cancer for many decades and, more recently, its value as an adjuvant treatment for premenopausal women with early-stage disease has been clearly demonstrated. This review examines the different methods of achieving ovarian ablation and assesses the relevance of ovarian suppression as a treatment aim. METHODS: Medline searches were used to identify recent key articles relating to the adenoma carcinoma sequence. Further relevant articles were obtained by manual scanning of the reference lists of selected papers. RESULTS AND CONCLUSION: Ovarian ablation has historically been achieved by surgical or radiological intervention. Although beneficial in premenopausal disease, these methods produce permanent ablation that results in a premature menopause, which may be associated with long-term adverse events. A series of trials have recently demonstrated the benefits of luteinizing hormone releasing hormone (LHRH) agonists, such as goserelin and leuprorelin, as adjuvant treatment for premenopausal hormone-sensitive disease. LHRH agonists produce a reliable suppression of ovarian oestrogen production of equivalent efficacy to adjuvant chemotherapy in hormone-sensitive disease. Effective ovarian suppression is marked by amenorrhoea, but studies have suggested that permanent amenorrhoea is not necessary to confer benefit in the treatment of early breast cancer. LHRH agonists therefore represent a beneficial therapeutic option for premenopausal patients with hormone-sensitive early disease. PMID- 12734856 TI - Systematic review of the quality of surgical mortality monitoring. AB - BACKGROUND: Mortality is the most tightly defined and used adverse event for audit and performance monitoring in surgery. However, to identify cause and therefore scope for improvement, accurate and timely data are required. The aim of this study was to perform a systematic review of the quality of measurement, reporting and monitoring of mortality as an outcome after surgery. METHODS: A systematic review of published literature was undertaken for the 7-year interval 1993-1999. Grey and unpublished literature was obtained through the Royal College of Surgeons of England, from UK national audits and routine national hospital data collections. RESULTS: Eligible monitoring systems included six UK national surgical audits, and cardiac and vascular surgery monitoring systems from North America and the UK. The definitions of 'surgical death' varied in several respects and deaths after discharge from hospital were rarely ascertained unless there was routine linkage to national death registers. There were very few published studies on validation of the completeness and accuracy of the data collection. CONCLUSION: A comprehensive data collection system is needed for improving clinical performance, with ownership, but not necessarily data collection, resting with the surgeons concerned. Recording of risk factors and deaths after discharge from hospital is essential, whatever data collection system is used. PMID- 12734857 TI - Pathophysiology of intra-abdominal adhesion and abscess formation, and the effect of hyaluronan. AB - BACKGROUND: Intra-abdominal adhesions and abscesses cause significant morbidity and mortality. The formation of fibrin in the abdominal cavity is a common pathophysiological pathway for both. The aim of this review was to investigate the pathophysiology of intra-abdominal adhesions and abscesses, and to explore the possible sites of action of hyaluronan. METHODS: Data were reviewed from the literature using the Medline database. RESULTS: Both surgery and peritonitis disturb the equilibrium between coagulation and fibrinolysis in the abdominal cavity in favour of the coagulation system. Hyaluronan-based agents reduce adhesion formation after surgery. Moreover, hyaluronan solution reduces abscess formation in experimental peritonitis. Possible mechanisms of action include mechanical separation of wound surfaces, improvement of peritoneal healing, modulation of the inflammatory response and enhanced fibrinolysis. CONCLUSION: Diminished fibrin degradation is a common pathway for the formation of adhesions and abscesses. The potential of hyaluronan-based agents to reduce intra-abdominal adhesions and abscesses in abdominal surgery and sepsis is a promising new concept. Elucidating the mechanisms involved and the clinical application of hyaluronan in peritonitis are challenges for future research. PMID- 12734859 TI - Enoxaparin effect depends on body-weight and current doses may be inadequate in obese patients. PMID- 12734858 TI - Randomized clinical study of Gastrografin administration in patients with adhesive small bowel obstruction. AB - BACKGROUND: Oral Gastrografin has been used to differentiate partial from complete small bowel obstruction (SBO). It may have a therapeutic effect and predict the need for early surgery in adhesive SBO. The aim of this study was to determine whether contrast examination in the management of SBO allows an early oral intake and reduces hospital stay. METHODS: Eighty-three patients admitted between February 2000 and November 2001 with 90 episodes of symptoms and signs suggestive of postoperative adhesive SBO were randomized into two groups, a control group and Gastrografin group. Patients in the control group were treated conservatively. If symptoms of strangulation developed or the obstruction did not resolve spontaneously after 4-5 days, a laparotomy was performed. Patients in the Gastrografin group received 100 ml Gastrografin. Those in whom the contrast medium reached the colon in 24 h were considered to have partial SBO, and were fed orally. If Gastrografin failed to reach the colon and the patient did not improve in the following 24 h a laparotomy was performed. RESULTS: Conservative treatment was successful in 77 episodes (85.6 per cent) and 13 (14.4 per cent) required operation. Among patients treated conservatively, hospital stay was shorter in the Gastrografin group (P < 0.001). All patients in whom contrast medium reached the colon tolerated an early oral diet. Gastrografin did not reduce the need for operation (P = 1.000). No patient died in either group. CONCLUSION: Oral Gastrografin helps in the management of patients with adhesive SBO and allows a shorter hospital stay. PMID- 12734861 TI - High recurrence rate after atypical resection for pancreatic metastases from renal cell carcinoma. AB - BACKGROUND: Pancreatic metastases from renal cell carcinoma are rare but highly resectable. The aim of this study was to review a series of patients with this condition. METHODS: The study involved 22 consecutive patients with histologically proven pancreatic metastases from renal cell cancer. RESULTS: Seventeen of the 22 patients had surgery. No patient died but eight of the 17 patients had a postoperative complication. Median follow-up was 33 (range 1-96) months. The 24- and 60-month survival probabilities were 0.84 and 0.53 respectively. Five patients who did not undergo surgery had 24- and 60-month survival probabilities of 0.53 and 0.26 respectively. The difference between the two groups was significant (P = 0.040). CONCLUSION: Despite the slow development of these secondaries and their well encapsulated morphology, the high rate of recurrence after limited resection suggests that radical resection should be recommended. PMID- 12734860 TI - 111In-labelled octreotide binding by the somatostatin receptor subtype 2 in neuroendocrine tumours. AB - BACKGROUND: The aim of this study was to investigate the importance of somatostatin receptor subtype 2 (SSTR2) expression for 111In-labelled diethylenetriamine-pentaacetic acid (DTPA)-D-Phe1-octreotide binding and uptake of 111In in neuroendocrine tumours. METHODS: 111In activity concentrations in surgical biopsies from neuroendocrine tumours (midgut carcinoid and medullary thyroid carcinoma), breast carcinoma and blood were determined 1-8 days after intravenous injection of 111In-labelled DTPA-D-Phe1-octreotide (140-350 MBq). The ratio of 111In activity concentrations between tumour tissue and blood (T/B value) was calculated. The expression of SSTR2 messenger RNA (mRNA) in tumour biopsies was quantitated by ribonuclease protection assay and SSTR2 protein was localized by immunocytochemistry. RESULTS: T/B values were highest for tumour biopsies from midgut carcinoids (mean 160 (range 4-1200); n = 65) followed by medullary thyroid carcinoma (mean 38 (range 2-350); n = 88) and breast carcinoma (mean 18 (range 4-41); n = 4). The expression of SSTR2 mRNA (relative to the NCI H69 cell line) was highest in tumour biopsies from midgut carcinoids (mean 2.5 (range 0.83-6.0); n = 40) followed by medullary thyroid carcinoma (mean 1.3 (range 0.20-6.0); n = 7) and breast carcinoma (mean 0.66 (range 0.29-1.0); n = 9). In tumour biopsies SSTR2 protein was localized exclusively to tumour cells. CONCLUSION: Midgut carcinoid tumours showed a much higher level of SSTR2 expression than medullary thyroid carcinoma in accordance with superior tumour imaging by octreotide scintigraphy. The high SSTR2 mRNA values and T/B values observed in midgut carcinoid tumours were positively correlated. PMID- 12734862 TI - Day-case laparoscopic Nissen fundoplication. AB - BACKGROUND: The aim was to assess the acceptability and safety of day-case laparoscopic fundoplication for gastro-oesophageal reflux disease (GORD). METHODS: This prospective study commenced in December 1999 and lasted for 18 months. All patients had proven symptomatic GORD. Inclusion criteria were American Society of Anesthesiologists grade I or II with adequate home support. A standard anaesthetic, analgesic and antiemetic protocol was used. Patients were contacted by telephone on the night of discharge and arrangements were made for a nurse to visit the following day. Postoperative pain and nausea were assessed using visual analogue scores (scale 0-10) on a self-completion questionnaire. RESULTS: Twenty patients were included. There were no postoperative complications. All patients were discharged on the day of surgery. Median time to discharge was 6 h 30 min (range 4.5 to 9 h). One patient reattended casualty the following morning but none required readmission. There was no significant difference in median pain or nausea scores the evening after surgery or the next morning. All patients were satisfied with the information given and aftercare provided. All would recommend it to a friend and 19 of 20 would undergo the procedure as a day case again. CONCLUSION: This study suggests that day-case laparoscopic fundoplication is feasible. Patients find it acceptable and it appears safe. PMID- 12734863 TI - Rapid immunohistochemical detection of lymph node micrometastasis during operation for upper gastrointestinal carcinoma. AB - BACKGROUND: The intraoperative diagnosis of lymph node micrometastasis (LNM) may help guide the area of appropriate lymph node dissection. This study aimed to evaluate the rapid immunohistochemical detection of LNMs using frozen sections during operation for gastro-oesophageal cancer. METHODS: Rapid immunostaining with anticytokeratin (AE1/AE3) antibody was compared with conventional immunostaining. A total of 210 lymph nodes obtained from 47 patients with oesophageal squamous cell carcinoma and from 32 with gastric adenocarcinoma were examined during operation. Lymph nodes were frozen, sectioned, and examined by histological and immunohistochemical methods. RESULTS: It took 30 min to complete the rapid immunostaining procedure; the expression of cytokeratin by rapid immunostaining was similar to that by conventional immunostaining. The incidence of lymph node metastasis detected by histological and immunohistochemical examination was 17 and 23 per cent respectively. LNM was solely detected in 12 lymph nodes by immunostaining: three micrometastases and nine with tumour cell microinvolvement. CONCLUSION: : Intraoperative rapid immunostaining is a simple and useful technique for detecting LNMs. Further study should investigate the role of rapid immunostaining during cancer surgery to select appropriate areas for lymphadenectomy. PMID- 12734864 TI - Extrahepatic disease does not contraindicate hepatectomy for colorectal liver metastases. AB - BACKGROUND: Extrahepatic disease has always been considered an absolute contraindication to hepatectomy for liver metastases. The present study reports the long-term outcome and prognostic factors of patients undergoing resection of extrahepatic disease simultaneously with hepatectomy for liver metastases. METHODS: From January 1987 to January 2001, 111 (30 per cent) of 376 patients who had hepatectomy for colorectal liver metastases underwent simultaneous resection of extrahepatic disease with curative intent. RESULTS: Surgery was considered R0 in 77 patients (69 per cent) and palliative (R1 or R2) in 34 patients (31 per cent). The mortality rate was 4 per cent and the morbidity rate 28 per cent. After a median follow-up of 4.9 years, the overall 3- and 5-year survival rates were 38 and 20 per cent respectively. The 5-year overall survival rate of patients with R0 resection only (n = 75) was 29 per cent. The difference in survival between patients with and without extrahepatic disease discovered incidentally at operation was significant, as was the number of liver metastases. CONCLUSION: Extrahepatic disease in patients with colorectal cancer who also have liver metastases should no longer be considered an absolute contraindication to hepatectomy. However, the presence of more than five liver metastases and the incidental intraoperative discovery of extrahepatic disease remain contraindications to hepatic resection. PMID- 12734865 TI - Use of transpelvic rectus abdominis musculocutaneous flap for anal cancer salvage surgery. AB - BACKGROUND: Perineal wounds following abdominoperineal resection (APR), for persistent or locally recurrent anal cancer, are associated with poor healing secondary to irradiation therapy. The results of APR combined with a vertical rectus abdominis musculocutaneous (VRAM) flap transposed transpelvically to cover the perineal defect are presented. METHODS: Between 1994 and 2000, 105 patients were diagnosed and treated for anal cancer. Twenty-two (21 per cent) underwent a salvage operation owing to persistent or locally recurrent disease. In eight patients, before 1996, the perineum was closed primarily with serious wound complications in five. In the final 14 patients, primary perineal reconstruction with a VRAM flap was performed. RESULTS: Median age was 65.5 (range 45-78) years. Median follow-up was 14.5 (range 3-41) months. There were no flap-related complications and primary healing was achieved in all patients. Median hospital stay was 17 (range 14-72) days. There were two major complications related to the laparotomy and abdominal closure. CONCLUSION: Combining the salvage operation with a VRAM flap facilitates primary healing after surgical treatment for persistent or locally recurrent anal cancer. A single-stage primary reconstructive procedure is feasible, with an acceptable complication rate and high level of patient satisfaction. PMID- 12734866 TI - Comparison of transanal stent with defunctioning stoma in low anterior resection for rectal cancer. PMID- 12734867 TI - Evidence of the effect of 'specialization' on the management, surgical outcome and survival from colorectal cancer in Wessex. AB - BACKGROUND: A prospective audit of the management of colorectal cancer was established to investigate factors associated with variation in survival observed within the former Wessex region (population three million). METHODS: Some 5173 patients (4562 surgically treated) with colorectal cancer diagnosed between 1991 and 1994 were followed for 5 years. Details of referral, diagnosis, surgical treatment, postoperative complications and outcomes were collected. The association between surgical outcomes and survival and both case volume and specialization (defined to include membership of the Association of Coloproctology of Great Britain and Ireland) was explored, accounting for variables with prognostic significance. RESULTS: There was a statistically significant association between high-volume operators (more than 50 operations per year) and specialization. The greatest benefit was observed with respect to specialists versus non-specialists, in terms of a lower postoperative mortality rate (odds ratio 0.67 (95 per cent confidence interval (c.i. 0.53 to 0.84)), lower anastomotic leak rates (odds ratio 0.46 (c.i. 0.31 to 0.66)), higher local recurrence-free survival (hazard ratio 0.56 (0.44 to 0.71)) and better long-term survival (hazard ratio 0.76 (c.i. 0.71 to 0.83)). CONCLUSION: There is a stronger association between surgical specialization in coloproctology and beneficial outcome than with high-volume caseloads. This is not entirely accounted for by case-mix or patient population, and is seen following colonic and rectal surgery and among patients with advanced disease. PMID- 12734868 TI - Radial artery as a conduit for diabetic crural bypass (Br J Surg 2003; 90: 57 58). PMID- 12734869 TI - The Physiological and Operative Severity Score for the enUmeration of Mortality and morbidity (POSSUM) (Br J Surg 2003; 90: 157-165). PMID- 12734870 TI - Mortality, morbidity and functional outcome after ileorectal anastomosis (Br J Surg 2003; 90: 59-65). PMID- 12734871 TI - Adjuvant therapy for rectal cancer cannot be based on the results of other surgeons (Br J Surg 2002; 89: 946-947). PMID- 12734872 TI - Randomized clinical trial comparing quality of life after straight and pouch coloanal reconstruction (Br J Surg 2003; 89: 1108-1117). PMID- 12734873 TI - Effects of the intermittent Pringle manoeuvre on hepatic gene expression and ultrastructure in a randomized clinical study (Br J Surg 2003; 90: 183-189). PMID- 12734874 TI - Effects of the intermittent Pringle manoeuvre on hepatic gene expression and ultrastructure in a randomized clinical study (Br J Surg 2003; 90: 183-189). PMID- 12734877 TI - Managing comorbidities in the rheumatic diseases: the new reality. PMID- 12734878 TI - Cases of early inflammatory polyarthritis should not be classified as having rheumatoid arthritis. PMID- 12734879 TI - Can progression of radiographic joint damage be predicted early in rheumatoid arthritis? PMID- 12734880 TI - Clinical evaluation versus ultrasonography: who is the winner? PMID- 12734881 TI - The use of biochemical markers of bone and cartilage metabolism to monitor osteoarthritis--dreams and reality. PMID- 12734882 TI - Corticotropin-releasing hormone promoter polymorphisms in patients with rheumatoid arthritis from northwest Spain. AB - OBJECTIVE: To investigate the possible implications of polymorphism in the CRH promoter in rheumatoid arthritis (RA) susceptibility, we examined a series of patients with RA from a defined area of Northwest Spain. METHODS: A total of 177 patients with RA and 147 ethnically matched controls from the Lugo region of Northwest Spain were studied. Patients and controls were genotyped for CRH polymorphisms in the 5' regulatory region of the gene at position 1273 (alleles A1 and A2) and at position 225 (alleles B1 and B2) by PCR-restriction fragment length polymorphism. Patients were stratified for age at onset of disease and rheumatoid factor status. RESULTS: When the whole group of patients was examined, no significant differences in CRH allele or genotype frequency were found compared with controls. However, the CRH allele A2 was significantly increased in patients with late onset seronegative RA compared with the seronegative group with younger age of disease onset (p = 0.03). In addition, 4 (36.4%) of the 11 patients with late onset seronegative RA carried the CRH-A2 allele versus only 2 (6.6%) of 31 patients with seronegative RA beginning before age 61 (OR 8.3, 95% CI 1.4-47.0; p = 0.015). CONCLUSION: In Northwest Spain, polymorphism in the CRH gene regulatory region may play a role as a disease susceptibility marker for late onset seronegative RA. PMID- 12734883 TI - Longterm protection of mice against collagen-induced arthritis after short-term LF 15-0195 treatment: modulation of B and T lymphocyte activation. AB - OBJECTIVES: LF 15-0195 is an immunosuppressive agent obtained by organic synthesis, currently under clinical development for the treatment of vasculitis. We define the effects of LF 15-0195 in the murine collagen-induced arthritis (CIA) model, an experimental model of human rheumatoid arthritis. METHODS: In our model, CIA was elicited in DBA/1 mice by immunization with bovine type II collagen (CII) in Freund's complete adjuvant, followed by a repeat injection 21 days later. Disease onset was observed 6 days after booster injection. In these experiments, mice were treated with 5 daily LF 15-0195 injections starting after the booster injection (days 21-25). The mice were observed for 40 days after the start of treatment, during which time arthritis was scored using clinical score and paw swelling assessment. Modulation of humoral immunity was documented by measuring the serum level of anti-CII IgG1 and IgG2a and cellular immunity by cytokines production by lymph node cells (LNC) and their proliferation in vitro. RESULTS: Short-term treatment of LF 15-0195 after booster injection prevented longterm development of CIA. LF 15-0195 inhibited B cell differentiation with a marked suppression of anti-CII IgG1 and IgG2a synthesis. Functional analyses of T lymphocytes showed that LF 15-0195 treatment reduces cytokine production by LNC after CII, anti-CD3, lipopolysaccharide stimulation. CONCLUSION: LF 15-0195 treatment during a short time period prevented development of arthritis, inhibited humoral-specific response longterm, induced a decrease in the number of LNC, and decreased cytokine production of T LNC after ex vivo stimulation. PMID- 12734884 TI - Role of Fcgamma receptors IIA, IIIA, and IIIB in susceptibility to rheumatoid arthritis. AB - OBJECTIVE: To investigate the role of Fcg receptor (FcgR) genes in susceptibility to rheumatoid arthritis (RA) using family based studies, to examine possible interactions between FcgR genotypes and the shared epitope (SE), and to assess linkage disequilibrium between FcgR loci. METHODS: Association studies were performed in 95 Caucasian, single-case, nuclear Caucasian families with both parents alive using haplotype based haplotype relative risk (HHRR) and transmission disequilibrium test (TDT) statistics. Three FcgR polymorphisms (FcgRIIA-131H/R, FcgRIIIA-158V/F, and FcgRIIIB-NA1/NA2) were genotyped using polymerase chain reaction methods. Linkage analysis was performed using 3 microsatellite markers (D1S498, D1S2844, D1S2762) flanking the FcgR region in an independent set of 90 Caucasian, multiple-case families. Potential effects of disease heterogeneity, including sex and the presence of rheumatoid factor, SE, and erosive or nodular disease, were taken into account in the analysis. Logistic regression analysis was performed to determine whether FcgR alleles are independent risk factors for the susceptibility to and/or severity of RA. Linkage disequilibrium was calculated using pairwise linkage disequilibrium statistics. RESULTS: HHRR and TDT analysis showed no evidence of preferential transmission of any FcgR alleles studied, and there were no important associations with any given disease phenotype. Moreover, neither linkage to microsatellite markers close to the FcgR genes on chromosome 1 nor linkage disequilibrium between FcgR loci was present in our population. The distribution of inherited genotypes provided evidence for an interaction between the SE and the FcgRIIIA-158V allele and between the SE and the FcgRIIIA-158V-FcgRIIA-131H 2-locus haplotype since the combined presence of these factors increased the susceptibility to RA (OR 4.13, 95% CI 1.6-10.62 and OR 2.83, 95% CI 1.25-6.38, respectively). However, regression analysis showed that neither the 158V allele nor the 158V-131H haplotype contributed as independent factors to susceptibility or severity of RA. CONCLUSION: Isolated FcgR genes do not play a major independent role in susceptibility to RA. To a limited extent, the presence of high-binding alleles at the FcgRIIIA locus or at the FcgRIIIA-FcgRIIA haplotype might predispose to RA in SE positive individuals. PMID- 12734885 TI - Therapeutic implications for interferon-alpha in arthritis: a pilot study. AB - OBJECTIVE: To evaluate the therapeutic potential of interferon-a (IFN-a) in osteoarthritis (OA) and rheumatoid arthritis (RA) by examining regulation of cytokine antagonist expression. METHODS: Expression of interleukin 1 receptor antagonist (IL-1Ra) and soluble tumor necrosis factor receptor (sTNFR) was examined by ELISA in cells from freshly isolated synovial fluids (SF) and synovial tissues (ST) from patients with OA or RA, either left untreated or treated with IFN-a. Single (7) and paired (5) SF and ST cells from OA and RA patients were examined. As well, the ability of IFN-a to regulate gene expression levels for osteoprotegerin (OPG) and osteoprotegerin ligand (OPGL) was examined in freshly isolated SF cells from patients with RA, by reverse transcriptase polymerase chain reaction. RESULTS: IL-1Ra and sTNFR were found to be constitutively expressed in OA and RA SF and ST cells. IFN-a treatment resulted in an increase in both IL 1Ra and sTNFR production. Freshly isolated RA SF cells exhibited constitutive OPGL gene expression in both the non-T and T cell fractions of the SF. In contrast, OPG gene expression levels were undetectable or low. IFN-a treatment of RA SF cells resulted in upregulation of OPG gene expression in the T cell fraction of the RA SF cells, whereas OPGL gene expression remained unaffected. CONCLUSION: These in vitro data suggest a therapeutic role for IFN-a in the treatment of arthritis through upregulation of critical cytokine antagonists. PMID- 12734886 TI - The responsiveness of generic health status measures as assessed in patients with rheumatoid arthritis receiving infliximab. AB - OBJECTIVE: We used a variety of health status measures in 2 groups of patients with rheumatoid arthritis (RA) to assess both the smallest distinguishable difference and the relative responsiveness to change of these measures, when used in clinical practice. METHODS: Two groups of patients were studied. Group 1: 24 patients with stable RA tested on 2 occasions; Group 2: 60 patients receiving methotrexate tested before and 14 weeks after treatment with infliximab. Assessments were made with self-completed questionnaires: the modified Health Assessment Questionnaire, Medical Outcomes Study Short Form-36 [SF-36 (SF-6D)], EuroQol, and, in some, the standard gamble. Group 2 also had joint counts, and measures of erythrocyte sedimentation rate, C-reactive protein, and hemoglobin. RESULTS: The limits-of-agreement (Bland-Altman) approach had greater confidence intervals (CI) than did CI based on +/- 2 standard errors of the measurement. Improvement with infliximab could be determined with all measures, however, but the standard gamble seemed least responsive to change. CONCLUSION: The various measures had different degrees of responsiveness, but with all it was possible to show improvement in Group 2 compared to Group 1. There was a closer association of the patient centered measures of improvement with changes in pain score than with joint counts. PMID- 12734887 TI - Prevalence of secondary amyloidosis in Asian North Indian patients with rheumatoid arthritis. AB - OBJECTIVE: To study the prevalence of secondary amyloidosis in Asian North Indian patients with rheumatoid arthritis (RA) and to determine its clinical significance. METHODS: RA patients with disease duration > 5 years were included in this prospective study over a 2 year period. Abdominal subcutaneous fat pad aspiration (ASFA) was performed, and smears were stained with Congo red and observed for apple-green birefringence under polarized light microscopy. The amyloid deposits were graded from 1+ to 3+. Clinical, radiological, and laboratory variables of the patients were correlated with the presence or absence of amyloidosis. RESULTS: Thirty out of 113 patients were positive for amyloid by ASFA (26.5%). Out of these, 8 patients had features suggestive of clinical amyloidosis in the form of proteinuria, organomegaly, or symptomatic gastrointestinal involvement. In another 22 patients amyloidosis was subclinical. The majority of patients with clinical amyloidosis had either 2+ or 3+ deposits. CONCLUSION: Abdominal fat amyloid deposits are not uncommon in adult Asian North Indian patients with RA. However, only one-fourth of patients had evidence of clinical amyloidosis. A longer followup and a larger multicentric collaborative study is needed to determine the significance of subclinical amyloid deposits. PMID- 12734888 TI - Health related quality of life in women with elderly onset rheumatoid arthritis. AB - OBJECTIVE: To examine the association of elderly onset rheumatoid arthritis (RA) with health related quality of life in a population based cohort of older women. METHODS: A nested case-control study of elderly onset RA within the Iowa Women's Health Study (IWHS), a prospective cohort established in 1986 of 41,000 women aged 55 to 69 years. A supplemental questionnaire was mailed to 122 RA cases and 1132 frequency matched controls from the cohort. We used unconditional logistic regression and linear regression to examine the association of elderly onset RA with self-reported measures of functional disability and quality of life. RESULTS: Elderly onset RA was associated with a 6-fold risk (OR 6.0, 95% CI 3.6 10.1) of significant functional disability (Health Assessment Questionnaire score (3) 1). Similarly, elderly onset RA was significantly associated with lower physical component scores of the Medical Outcome Study Short Form-12 (37.2 +/- 10.9 vs 43.6 +/- 11.6; p < 0.001). CONCLUSION: Among a community based cohort, elderly onset RA was strongly associated with functional disability and reduced quality of life. These associations were independent of other age associated factors including depression, recent fracture, and multiple comorbidities. PMID- 12734889 TI - National study of cause-specific mortality in rheumatoid arthritis, juvenile chronic arthritis, and other rheumatic conditions: a 20 year followup study. AB - OBJECTIVE: To quantify risks for cause-specific mortality among hospitalized patients with rheumatoid arthritis (RA), juvenile chronic arthritis (JCA), and 4 other rheumatic conditions in a nationwide, population based cohort over a 20 year period. METHODS: All subjects were identified from Scottish hospital inpatient records from 1981 to 2000 and were followed up by computer linkage to the national registry of deaths. Expected mortality was calculated from national mortality rates and was related to the observed incidence by the standardized mortality ratio (SMR) and the corresponding 95% confidence interval (95% CI). RESULTS: Overall mortality was elevated in each of the 6 rheumatic conditions examined, most notably in JCA (males: SMR 3.4, 95% CI 2.0,5.5; females: SMR 5.1, 95% CI 3.2,7.8). Among patients with RA, there was an increased risk for death in all International Classification of Disease chapters other than those relating to mental disorders. Specific causes of death with an increased risk for subjects with RA included lung cancer [males: 1.4 (1.2,1.5); females: 1.6 (1.5,1.8)], hematopoietic malignancies [M: 1.8 (1.4,2.3); F: 2.0 (1.7,2.3)], coronary artery disease (CAD) [M: 1.6 (1.5,1.7); F: 1.95 (1.9,2.0)], respiratory infections [M: 1.9 (1.7,2.2); F: 2.4 (2.3,2.6)], chronic obstructive pulmonary disease [M: 1.8 (1.6,2.0); F: 2.1 (1.9,2.3)], and renal failure [M: 3.1 (2.5,3.9); F: 3.5 (3.0,4.0)]. Conversely, RA subjects were less likely to die from gastrointestinal tract malignancies [M: 0.82 (0.7,1.0); F: 0.8 (0.7,0.9)]. CONCLUSION: Population studies for primary data collection are required to extend our knowledge about the underlying mechanisms of early mortality in patients with rheumatic conditions. PMID- 12734890 TI - Ultrasonography is superior to clinical examination in the detection and localization of knee joint effusion in rheumatoid arthritis. AB - OBJECTIVE: Musculoskeletal ultrasonography allows real-time imaging of joint structures and may be used to complement clinical examination in rheumatological practice. We compared ultrasonography (US) with clinical examination (CE) in the detection of effusion, suprapatellar bursitis, and Baker's cyst of the knee in rheumatoid arthritis (RA) in order to determine whether US provided additional clinical information. METHODS: A total of 22 patients with RA (ACR criteria) underwent independent clinical and US examination of both knees for suprapatellar bursitis, knee effusion, and presence of Baker's cyst. US was performed using an ATL HDI 3000 machine with L7-4 MHz and CL10-5 MHz probes. Clinical examination was performed using standard techniques by an experienced rheumatologist. Patients with previous knee surgery were excluded from the study. RESULTS: A total of 44 knees were examined at a total of 130 sites (one patient was unable to lie prone for US of popliteal fossae). US detected soft tissue abnormality (suprapatellar bursitis, knee effusion, or Baker's cyst) at 54/130 (42%) sites, while CE detected soft tissue abnormality at 36/130 (28%) sites. US detected 17 (39%) cases of suprapatellar bursitis in 44 knees, 7 (16%) of which were detected on CE. US detected 27 (61%) knee joint effusions in 44 knees, 16 (36.36%) of which were detected on CE. US detected 10 (23.81%) Baker's cysts in 42 knees, 2 (4.76%) of which were detected on CE. Taking US of the knee as the gold standard, CE was specific but not sensitive in the detection of soft tissue abnormality of the knee in RA. CONCLUSION: US is more sensitive than CE in the detection of suprapatellar bursitis, knee effusion, and Baker's cyst in RA. CE underestimates knee inflammation in RA. This has implications for the use of CE as a component of standardized disease activity scores and in guiding knee joint aspiration. PMID- 12734892 TI - Design, quality, and bias in randomized controlled trials of systemic lupus erythematosus. AB - OBJECTIVE: To appraise systematically the study design and quality of reporting of randomized controlled trials (RCT) on systemic lupus erythematosus (SLE) and to identify potential defects and biases. METHODS: RCT with at least 5 patients with SLE were retrieved from MEDLINE, EMBASE, and the Cochrane Library. We analyzed study design, quality of reporting, and trial results. RESULTS: Ninety four trial reports (37 on lupus nephritis) were eligible with 2,257 SLE patients (n = 795 in lupus nephritis trials). Median sample size was 28 patients. Fifty one trials (54.3%) were double blind, but only 31 (33.0%) mentioned the randomization mode, only 19 (20.2%) described allocation concealment, and only 7 (7.5%) were adequately powered. Sixty-three trials (67%) described adequately reasons for withdrawals. Nephritis trials had on average longer followup (p = 0.001) and were less likely to be double blind (p < 0.001), to describe reasons for withdrawals [both overall (p = 0.008) and per arm (p = 0.009)] and to involve a comparison against placebo or no treatment (p < 0.001). Larger trials scored higher on several quality characteristics. Significant efficacy or trend for efficacy was claimed in 72 reports (76.6%) and this was even more common in trials published in 1999-2002 (89.5%). Significant efficacy was found more frequently in trials that clearly specified withdrawals per arm (p = 0.001) and outcomes (p = 0.001) and used intention-to-treat analyses (p = 0.03). Besides outcome specification, no other quality variables seemed to improve significantly over time. CONCLUSION: Several aspects of the design and reporting of RCT on SLE can be improved. Larger, adequately powered, and accurately reported trials are needed. PMID- 12734891 TI - Cytotoxicity responses to peptide antigens in rheumatoid arthritis and ankylosing spondylitis. AB - OBJECTIVE: To measure levels of IgG antibodies against structurally related synthetic peptides of HLA-DRB1*0404, type XI collagen, and Proteus mirabilis in patients with rheumatoid arthritis (RA) and HLA-B*2705 and Klebsiella pneumoniae in patients with ankylosing spondylitis (AS), and to determine whether sera from RA and AS patients are cytotoxic for sheep red blood cells (SRBC) coated with HLA DRB1*0404, type XI collagen, or HLA-B*2705. METHODS: Sera from 51 patients with RA, 34 with AS, and 38 healthy controls were tested against synthetic EQRRAA, ESRRAL, LRREI, and IRRET peptides by ELISA. Sera from patients and controls were also tested for reactivity in complement mediated cytotoxicity with SRBC coated with EQRRAA and HLA-B*2705, LRREI peptides. RESULTS: Antibodies to synthetic peptides containing EQRRAA, ESRRAL, LRREI, and IRRET were significantly increased in RA patients compared with AS patients (p < 0.001) and controls (p < 0.001). The percentage lysis data for SRBC coated with EQRRAA and LRREI peptides were significantly higher for RA sera (p < 0.001) compared to control sera. Percentage lysis for SRBC coated with HLA-B*2705 peptide was significantly higher for AS sera (p < 0.001) compared to control sera. CONCLUSION: Our results suggest that antibodies against antigenic determinants of P. mirabilis in RA and K. pneumoniae in AS have cytotoxic properties on structurally related host proteins. These cytotoxic antibodies together with T cell interactions could be relevant in the etiopathogenesis of RA and AS. PMID- 12734893 TI - Neuropsychiatric manifestations in systemic lupus erythematosus: prevalence and association with antiphospholipid antibodies. AB - OBJECTIVE: To apply the new American College of Rheumatology nomenclature for neuropsychiatric systemic lupus erythematosus (NPSLE), determine the prevalence of the different neuropsychiatric (NP) syndromes, and evaluate which of these manifestations correlates with the presence of antiphospholipid antibodies (aPL). Methods. Clinical, serological, and imaging data of 323 consecutive patients with SLE were retrospectively reviewed. Neuropsychometric testing was applied by a neuropsychologist. Univariate and multivariate statistical analyses were applied to evaluate the association bewteen NP manifestations, magnetic resonance imaging (MRI) abnormalities, and aPL. RESULTS: In total, 185 patients (57.3%) had NP manifestations at any time during followup. Headache was the most frequent manifestation, present in 78 patients (24%). Cerebrovascular disease (CVD) was diagnosed in 47/323 patients (14.5%), with a total of 57 events. Mood disorders were found in 54 (16.7%), cognitive disorders in 35 (10.8%), and seizures in 27 patients (8.3%). Psychosis was diagnosed in 25 (7.7%), anxiety disorder in 24 (3.7%), and acute confusional state in 12 patients (3.7%). Less common manifestations were polyneuropathy, mononeuritis, myasthenia gravis, cranial neuropathy, myelopathy, chorea, demyelinating disease, and Guillain-Barre syndrome. The presence of aPL was associated with NP manifestations (p < 0.001). Multivariate analysis showed that aPL were independently associated with CVD (OR 6.17, 95% CI 2.94-12.9, p = 0.001), headache (OR 2.04, 95% CI 1.17-3.55, p = 0.01), and seizures (OR 2.89, 95% CI 1.18-7.10, p = 0.02). The presence of lupus anticoagulant (LAC) was independently associated with white matter hyperintensity lesions on MRI (OR 3.0, 95% CI 1.12-8.05, p = 0.027). CONCLUSION: The new ACR criteria for NPSLE are useful to define NP manifestations in SLE with accuracy. NP manifestations are significantly associated with aPL. CVD, headache, and seizures were independently associated with these antibodies. PMID- 12734895 TI - Clinical significance of antibodies to TS1-RNA in patients with mixed connective tissue disease. AB - OBJECTIVE: To investigate the clinical significance of anti-TS1-RNA antibodies in patients with mixed connective tissue disease (MCTD). METHODS: Anti-TS1-RNA antibodies were detected by immunoprecipitation using 32P-UTP labeled TS1-RNA as the antigen source. In total, 104 patients with MCTD, 30 with Sjogren's syndrome, 30 with systemic lupus erythematosus (SLE), 25 with systemic sclerosis, 23 with polymyositis or dermatomyositis, and 10 with rheumatoid arthritis were examined. Specificity of anti-TS1-RNA antibodies was analyzed by immunoprecipitation using HeLa cell extracts. RESULTS: The frequency of anti-TS1-RNA antibodies was 31.7% in patients with MCTD, significantly higher than in SLE (p < 0.05). In anti-TS1 RNA positive patients, the incidence of hypertension and proteinuria and the frequency of anti-Sm and anti-dsDNA antibodies associated with SLE were higher than those of anti-TS1-RNA negative patients. Clinical features of SS such as sicca complex, the serum level of IgA, and anti-SSA antibodies were also elevated. The frequency of anti-TS1-RNA antibodies was significantly higher in SLE patients with anti-U1-RNP antibodies (p < 0.01); however, anti-TS1-RNA positive sera did not precipitate the specific RNA including U1 RNA in immunoprecipitation using HeLa cell extracts. In longitudinal studies, the level of anti-TS1-RNA antibodies changed in parallel with disease activity. CONCLUSION: We found that the level of anti-TS1-RNA antibodies was possibly correlated with the disease activity of lupus-like clinical features in patients with MCTD. PMID- 12734894 TI - Antiguanosine antibodies in murine and human lupus have the internal image of G binding proteins. AB - OBJECTIVE: To examine the binding specificities of serum IgG antibodies of mouse and human origin directed against guanosine. The immunodominance of guanosine compared with the other nucleosides was established in the MRL/lpr murine model of systemic lupus erythematosus (SLE). Serum antiguanosine autoantibodies in human lupus correlate with nephritis and polyserositis in acute disease as well as in exacerbations of disease symptoms. METHODS: Antiguanosine autoantibodies obtained from humans with SLE were compared to a murine monoclonal antiguanosine antibody, 4H2. The fine specificity of the antiguanosine-binding site was determined by methylation of specific positions on the guanosine molecule and using defined analogs in competitive ELISA. RESULTS: Competitive inhibition assays revealed that serum antiguanosine antibodies bind across the 1 and 7 positions of the guanosine molecule (p < 0.01) and that an oxygen is necessary at position 6 in the molecule. 4H2 exhibited the same binding specificity for guanosine as human polyclonal antiguanosine antibodies, showing a conserved epitope across species. When the fine specificity was compared with known epitopes, the antiguanosine antibodies were found to have the internal image of a G-binding protein, identical to that of the Ha-ras oncogene product p21. CONCLUSION: The finding that antiguanosine autoantibodies vary directly with specific features of SLE, especially nephritis and polyserositis, suggests that they may contribute to the pathology of SLE. Our findings that antiguanosine antibodies have G-binding protein active site homology support the possibility that this species of antibody might interfere with cell signal transduction. PMID- 12734896 TI - How "soft" are soft neurological signs? The relationship of subjective neuropsychiatric complaints to cognitive function in systemic lupus erythematosus. AB - OBJECTIVE: As part of a longitudinal study of cognitive function in systemic lupus erythematosus (SLE), we documented the range and frequency of subjective neurologic and/or psychiatric (NP) complaints in Never-NP-SLE patients, and related these to cognitive function, using the latter as a primary indicator of nervous system involvement. METHODS: Thirty patients with SLE who did not have major neurologic and psychiatric involvement underwent baseline and followup neuropsychological testing roughly 5 years apart. Within 0-13 months prior to retesting, each patient completed a 42 item questionnaire recording NP symptoms. RESULTS: The group as a whole endorsed 26% of symptoms. Fourteen patients labelled high endorsers (> 35% of items) endorsed, on average, 42% of symptoms. There was a significant association between higher item endorsement and lower cognitive function (r = -0.46, p < 0.02) and significantly poorer cognitive performance in the high compared to low endorser groups (t = -3.07, p < 0.005). In addition, a subset of 8 items was endorsed at least twice as often by SLE patients as by patients with rheumatoid arthritis (n = 12) or healthy controls (n = 10). CONCLUSION: These results suggest that "minor" NP symptoms and, in particular, a small subset of subjective complaints may be sufficient to raise suspicion of subclinical nervous system involvement in the absence of clinically evident NP-SLE. PMID- 12734897 TI - Group psychotherapy reduces illness intrusiveness in systemic lupus erythematosus. AB - OBJECTIVE: We investigated whether brief supportive-expressive group psychotherapy might reduce illness-induced interference with valued activities and interests (i.e., illness intrusiveness) among women with systemic lupus erythematosus (SLE) in relation to 3 life domains: (1) relationships and personal development (family relationships, other social relationships, self-expression), (2) intimacy (relationship with spouse, sex life), and/or (3) instrumental life (work, finances, active recreation). METHODS: Women with SLE recruited from 9 rheumatology centers were randomly assigned to receive either usual care (n = 66) or a 12 week brief supportive-expressive group psychotherapy followed by 3 monthly booster sessions (n = 58). Standard instruments assessed disease activity and damage, illness intrusiveness, and psychological distress at 4 measurement occasions: (1) pretreatment, (2) posttreatment, (3) 6 month followup, and (4) 12 month followup. RESULTS: Analysis of covariance, controlling for disease activity and household income, indicated that women who received brief supportive expressive group psychotherapy experienced significant reductions in illness intrusiveness for 2 of 3 domains: (1) relationships and personal development and (2) intimacy. Benefits were evident at 6 and 12 month followups. CONCLUSION: Brief supportive-expressive group psychotherapy facilitates adaptation to SLE by assisting women in reducing illness-induced disruptions into important domains of life experience. PMID- 12734898 TI - Treatment of subglottic stenosis, due to Wegener's granulomatosis, with intralesional corticosteroids and dilation. AB - OBJECTIVE: To determine the longterm efficacy of intralesional long-acting corticosteroid injection plus dilatation (ILCD) for subglottic stenosis (SGS) in Wegener's granulomatosis (WG). METHODS: Since November 1994, all patients with WG who presented with SGS of more than 50% or symptoms of airway compromise were treated with intralesional injection of methylprednisolone acetate, injected directly into the stenotic segment, followed by microsurgical lysis of the stenotic ring and serial dilatation with Maloney bougies or Fogarty catheter balloon. The procedure was repeated at a later date if re-stenosis occurred. Patient outcome was evaluated over a period of 7 years. RESULTS: Twenty-one patients underwent 64 procedures. Mean followup was 40.6 months. Patients who did not have scarring from prior procedures required a mean of 2.4 procedures at mean intervals of 11.6 months to maintain subglottic patency. Patients with established laryngotracheal scarring required a mean of 4.1 procedures at mean intervals of 6.8 months to maintain patency. None of the 21 patients required a new tracheostomy. Only 2 significant complications occurred, both pneumothoraces. There were no adverse longterm sequelae. CONCLUSION: ILCD is effective therapy for SGS due to WG. Best results are obtained when these endoscopic techniques are performed prior to other forms of surgery, which may produce extensive scar formation. Based on this experience, the authors recommend ILCD as the preferred therapy in WG-SGS. PMID- 12734899 TI - Clinical and radiological changes during psoriatic arthritis disease progression. AB - OBJECTIVE: To determine the contribution of radiological findings at the initial visit with respect to classifying patients with psoriatic arthritis (PsA); and to determine the extent to which the clinical disease patterns change over time. METHODS: Patients with PsA were followed prospectively at 6-12 month intervals since 1978; 86 patients were registered within 1 year of diagnosis and were followed for at least 1 year. Based on the clinical information, including the actively inflamed joint count, damaged joint count, and the presence of back disease and arthritis mutilans, a clinical PsA pattern was assigned. A separate radiology pattern based on radiographs alone, and a combined clinical and radiological pattern, was also assigned at each visit. The initial clinical pattern was compared to the initial combined clinical-radiological pattern, and the initial clinical pattern was compared to the clinical pattern at 1 year and 5 years in 35 patients with both 1 and 5 year followup. RESULTS: In 23% of the patients, the radiological assessment in the initial visit showed evidence of patterns not detected clinically; 49% and 77% of the patients showed clinical pattern change within 1 year and 5 years, respectively. A comparison between the group that changed pattern and the one that did not change pattern in 5 years revealed no significant features. CONCLUSION: Radiological assessments add information not gained from clinical assessment alone. Clinical patterns do change over time in the majority of patients. Both elements must be taken into consideration when developing classification criteria for PsA. PMID- 12734900 TI - High ferritin and low glycosylated ferritin may also be a marker of excessive macrophage activation. AB - OBJECTIVE: A high serum ferritin concentration with a low percentage of glycosylated ferritin (< 20%) have been reported to be a specific marker of active adult Still's disease (ASD). However, high ferritin levels are found during hemophagocytosis syndrome (HS). We investigated the ferritin level and the percentage of glycosylation in a HS series of various causes. METHODS: Diagnosis of HS was confirmed by erythrophagocytosis pictures on a bone marrow cytology or biopsy in all patients. Serum ferritin concentration was determined on a heterogenous immunoassay module. Glycosylated ferritin was separated using concanavalin A (Con-A) sepharose 4B chromatography. The nonglycosylated ferritin unbound to Con-A was recovered in the supernatant and quantified with the same procedure. Percentages of glycosylated ferritin less than 20% are considered to be usual in ASD, between 20 and 40% usual in inflammatory syndrome, and between 50 and 80% normal. RESULTS: In all cases tested during the acute phase of the disease, ferritin blood level was high and the percentage of glycosylated ferritin was low, less than 20%. CONCLUSION: The combination of high ferritin level and low percentage of glycosylation may be a marker of excessive macrophage activation. PMID- 12734901 TI - Inter and intraobserver variability in DePalma's classification of shoulder calcific tendinitis. AB - OBJECTIVE: To determine the intraobserver reproducibility and interobserver reliability of DePalma and Kruper's classification of calcific tendinitis of the rotator cuff. The result of this classification influences the choice of therapeutic procedures in patients with symptomatic calcific tendinitis. METHODS: Plain anteroposterior radiographs of shoulders from 100 patients with symptomatic calcified deposits of the rotator cuff were classified according to the criteria of DePalma and Kruper by 6 independent observers at 2 different time points within 4 months. The kappa values of intraobserver reproducibility and interobserver reliability were calculated. RESULTS: Kappa values of intraobserver reproducibility had a mean of 0.487 (SD 0.094); kappa values of interobserver reliability were 0.234 for the first test and 0.273 for the second test. CONCLUSION: Determination of intraobserver reproducibility gave satisfactory to sufficient results and interobserver reliability was satisfactory for both tests indicating that studies based on the classification of DePalma and Kruper should be interpreted cautiously. PMID- 12734902 TI - The utility of alizarin red s staining in calcium pyrophosphate dihydrate crystal deposition disease. AB - OBJECTIVE: To determine the most suitable staining method for preservation and detection of calcium pyrophosphate dihydrate (CPPD) crystals in histological sections of patients with CPPD crystal deposition disease. METHODS: Paraffin sections of CPPD crystal-bearing tissues of 31 patients were stained with hematoxylin and eosin (H&E) and Alizarin red S (ARS). For H&E, the sections were treated with Mayer's hematoxylin (pH 2.3) for 5 min and with eosin alcohol (pH 4.1) for 1 min. For ARS, 1% ARS dissolved in distilled water was adjusted to pH 6.4 by adding 0.1% ammonia solution drop by drop while stirring. As controls, unstained sections were soaked in 1% citric acid monohydrate solution (CAMS, pH 2.3) for 5 or 10 min. The histological preparations were examined under a compensated polarized light using a first-order red compensator. We counted the number of weakly positive birefringent CPPD crystals in 3 high power fields (HPF, 0.272 mm2). RESULTS: CPPD crystals were seen clearly in most specimens stained with ARS, but were markedly reduced in tissue sections stained with H&E or CAMS. The number of CPPD crystals detected in sections stained by ARS (1723 +/- 683 per 3 HPF, mean +/- standard deviation) was significantly higher compared with H&E, CAMS (5 min), and CAMS (10 min) (401 +/- 374, 1022 +/- 616, and 494 +/- 636 per 3 HPF, respectively; p < 0.001, each). CONCLUSION: Standard H&E staining reduces the number of visible CPPD crystals, probably due to the strong acidity of both hematoxylin and eosin solutions, whereas the ARS stain seems to preserve a large number of CPPD crystals. The utility of ARS staining may improve the identification of CPPD crystals and contribute to a correct diagnosis of CPPD crystal deposition. PMID- 12734903 TI - Effect of exercise and beer on the plasma concentration and urinary excretion of purine bases. AB - OBJECTIVE: To investigate the effects of exercise and beer ingestion separately and combined on the plasma concentration of purine bases. METHODS: Six healthy men aged 30-39 years participated in 3 different experiments, in which they exercised for 30 min (at 70% of maximum oxygen uptake) and ingested beer (10 ml/kg body weight), or did each activity separately, with each experiment performed at 2 week intervals. RESULTS: The plasma concentration of uric acid was increased by 12% (p < 0.05), 8% (p < 0.01), and 29% (p < 0.01) with exercise, beer ingestion, and a combination of exercise and beer ingestion, respectively, which showed that it increased synergistically in the combination experiment. The fractional excretion of uric acid was decreased by 44% (p < 0.01) and 52% (p < 0.01) with exercise alone and a combination of exercise and beer ingestion, respectively, while it was increased by 15% (p < 0.05) with beer ingestion alone. Creatinine clearance was decreased by 16% (p < 0.01) with both exercise alone and a combination of exercise and beer ingestion, while it was not changed with beer ingestion alone. The increase in the plasma concentration of xanthine during the beer ingestion experiment was 2.1-fold greater than that during the combination (p < 0.05), while the increase in urinary excretion of xanthine caused by beer ingestion was 2.5-fold greater than that caused by a combination of beer and exercise (p < 0.05). Finally, exercise alone as well as a combination of beer and exercise increased the blood concentrations of lactic acid and NH3, whereas beer alone decreased concentration of pyruvic acid. CONCLUSION: These results suggest that the production of uric acid caused by both exercise and beer ingestion, as well as the inhibition of urinary uric acid excretion from a high blood lactic acid concentration, were the main contributors to the synergistic effect on the increase in plasma uric acid concentration. A decrease in creatinine clearance also contributed to the effect. We considered that pyruvic acid and NH3, produced in the muscles following exercise, relieved the beer induced increase of the plasma concentration and urinary excretion of xanthine, which may have played a minor role in the increase in plasma uric acid concentration. PMID- 12734904 TI - Biochemical markers of bone and cartilage remodeling in prediction of longterm progression of knee osteoarthritis. AB - OBJECTIVE: To investigate the relationship between biochemical markers of bone and cartilage remodeling and severity or progression (symptoms and structure) of knee osteoarthritis (OA). METHODS: Mean and minimal joint space width (JSW) of the femorotibial joint were measured from standardized radiographs taken at baseline and at the end of a 3-year longitudinal study of patients with knee OA. Pain, stiffness, and physical function subscales of the Western Ontario and McMaster Universities Osteoarthritis (WOMAC) index were assessed at the same time points. Biochemical markers [serum keratan sulfate (KS), serum hyaluronic acid (HA), urine pyridinoline (PYD) and deoxypyridinoline (DPD), serum osteocalcin (OC), cartilage oligomeric matrix protein (COMP)] were assessed at baseline and after 1 year. RESULTS: At baseline, no significant correlations were observed between values of biochemical markers and JSW or any of the WOMAC scores. Baseline markers were not correlated with 3-year percentage changes observed in mean or minimal JSW and WOMAC scores. Changes observed after 1 year in OC and HA were significantly correlated with 3-year progression in mean JSW (r = -0.24, p = 0.04 and r = 0.27, p = 0.02, respectively) and in minimal JSW (r = -0.31, p = 0.01 and r = 0.24, p = 0.04, respectively). In patients from the lowest quartile of 1-year changes in HA (< -21.22 ng/ml), mean JSW decreased after 3 years by 0.76 (1.23) mm compared to an increase of 0.11 (0.83) mm in patients in the highest quartile (> +14.34 ng/ml) (p = 0.03). CONCLUSION: The 3-year radiological progression of knee OA could be predicted by a 1-year increase in OC or a 1-year decrease in HA levels. PMID- 12734905 TI - HLA-B*39 allele confers susceptibility to osteoarticular complications in human brucellosis. AB - OBJECTIVE: To determine the contribution of HLA gene polymorphism toward susceptibility to osteoarticular focal forms of human brucellosis. METHODS: A total of 57 patients with brucellosis, of whom 23 had osteoarticular complications, and 73 healthy volunteers were genotyped for HLA class I and class II antigens by a polymerase chain reaction-sequence specific primer technique. RESULTS: The HLA-B*39 allele was present in significantly more patients with osteoarticular complications than in the other patients (35% vs 3%; p = 0.0006, OR 15.684, 95% CI 3.453-71.231), or in the controls. CONCLUSION: The increased presence of the HLA-B39 genotype in patients with brucellosis with clinical osteoarticular manifestations suggests that this genotype confers susceptibility to developing severe osteoarticular focal forms of the disease. PMID- 12734906 TI - Quality of life in women with fibromyalgia syndrome: validation of the QIF, the French version of the fibromyalgia impact questionnaire. AB - OBJECTIVE: To validate a translated and adapted version of the Fibromyalgia Impact Questionnaire (FIQ) for use in French-speaking populations. METHODS: The FIQ was translated into French by 2 independent translators and then back translated into English to assess the conceptual equivalence. The translated version was tested and adapted by an expert committee to obtain the Questionnaire de mesure d'Impact de la Fibromyalgie (QIF), the French version of the FIQ. We administered the QIF to 102 women with fibromyalgia (FM): 71 women who consulted once, and 31 women who were follow for 3 visits (D0, M1, and M3). The patients were also asked to answer 4 other questionnaires: the McGill Pain Questionnaire, the Medical Outcome Study Short Form-36 (SF-36), the short form of the Arthritis Impact Measurement Scale 2 (AIMS2), and the General Health Questionnaire (GHQ) (for psychiatric assessment). To ensure test-retest reliability, the patients were asked to complete the QIF 7 days after the first visit and to send it back to the investigators by mail. During each visit, all patients were asked about pain intensity. A tender point count was obtained by thumb palpation and the tenderness threshold of each specific point was assessed by a 4-point scale score to determine the global tender point index. RESULTS: No major cultural adaptation was needed to obtain the French version of the FIQ. Test-retest reliability coefficients (intraclass correlation coefficient) for each question ranged from 0.04 to 0.84. Two items from the QIF (number of days when the patient felt good and visual analog scale stiffness) did not reach significant levels of test retest reliability. Internal validity was good. The QIF score correlated well with the SF-36 and AIMS2 scores. The psychological aspects of the QIF were well correlated with those of GHQ-28. None of the items from the McGill Pain Questionnaire was correlated with QIF items. Similarly the clinical data concerning pain assessment were not correlated with QIF items. CONCLUSIONS: QIF is a valid instrument for measuring functional disability and health status in French women with FM. Some of the items were of a limited reliability, perhaps due to the variability of the multiple aspects of this syndrome. PMID- 12734907 TI - Aerobic fitness effects in fibromyalgia. AB - OBJECTIVE: To compare 2 exercise modalities, aerobic fitness training and stretching exercises, in patients with fibromyalgia (FM) in relation to function, pain, quality of life, depression, and anxiety, and to correlate the cardiorespiratory fitness gain with symptom improvement. METHODS: Seventy-six women with FM between 18 and 60 years old were randomized to either an aerobic program or stretching program, for 20 weeks. They were evaluated at the beginning of the program and after 10 and 20 weeks in relation to the improvement of aerobic fitness, flexibility, function, Fibromyalgia Impact Questionnaire (FIQ), Short-form Health Survey (SF-36), and depression and anxiety levels. Ventilatory anaerobic threshold (VT) and maximum oxygen uptake (VO2max) were determined by expired gas analyses. RESULTS: Aerobic exercise was superior to stretching in relation to VO2 max, VT, function, depression, pain, and the emotional aspects and mental health domains of SF-36. Patients in the stretching group showed no improvement in depression, "role emotional," and "mental health." No association was noted between improvement in aerobic fitness as measured by VT and the improvement of pain, function, or scores in FIQ and SF-36. CONCLUSION: Our results confirm that aerobic exercise is beneficial to patients with FM, but the cardiorespiratory fitness gain is not related to improvement of FM symptoms. PMID- 12734909 TI - Aerobic exercise capacity in patients with juvenile dermatomyositis. AB - OBJECTIVE: To examine the feasibility of maximum exercise testing in patients with juvenile dermatomyositis (JDM), characterize the maximum oxygen consumption (VO2peak) of these patients, and determine if exercise time could be used as an surrogate index for VO2peak. METHODS: Fifteen patients diagnosed with JDM (age 5 14) performed a graded, maximum exercise test using a motor driven treadmill and metabolic cart to volitional exhaustion conforming to the Bruce protocol. RESULTS: All patients were able to perform the exercise test. Ten of the 15 patients performed a maximal effort (heart rate > 180 beats/min or respiratory exchange ratio > 1.0). The patients who had a maximal exercise performance, the mean absolute VO2peak, relative VO2peak (related to body mass), and exercise time were respectively -1.82 (+/- 1.5), -2.83 (+/- 1.9), and -3.65 (+/- 1.9) standard deviations lower compared to age and sex matched reference values (p < 0.05). Z scores for exercise time were significantly lower compared to Z scores for absolute and relative VO2peak, an indication of reduced muscular economy. When exercise time was converted to VO2peak using the equation: VO2peak = 0.1583 (exercise time) + 0.0828, the VO2peak values were not significantly different from the measured VO2peak values. CONCLUSION: Aerobic exercise testing on a treadmill was possible in patients with JDM and revealed an impairment in their maximal aerobic exercise capacity. Exercise time can be used as an indicator of VO2peak, when converted to VO2peak using a regression equation. VO2peak, measured or estimated, has the potential to be a good indicator of muscle function in patients with JDM. PMID- 12734908 TI - The effects of sodium oxybate on clinical symptoms and sleep patterns in patients with fibromyalgia. AB - OBJECTIVE: Fibromyalgia (FM) is associated with the sleep phenomenon of alpha intrusion, and with low growth hormone secretion. Sodium oxybate has been shown to increase both slow-wave sleep and growth hormone levels. This double blind, randomized, placebo controlled crossover trial was conducted to evaluate the effects of sodium oxybate on the subjective symptoms of pain, fatigue, and sleep quality and the objective polysomnographic (PSG) sleep variables of alpha intrusion, slow-wave (stage 3/4) sleep, and sleep efficiency in patients with FM. METHODS: Patients received either 6.0 g/day sodium oxybate or placebo for 1 month, with an intervening 2 week washout period. Efficacy measures included PSG evaluations, tender point index (TPI), and subjective measurements from daily diary entries. Safety measures included clinical laboratory values, vital signs, and adverse events. RESULTS: Twenty-four female patients were included in the study; 18 completed the trial. TPI was decreased from baseline by 8.5, compared with an increase of 0.4 for placebo (p = 0.0079). Six of the 7 pain/fatigue scores (overall pain, pain at rest, pain during movement, end of day fatigue, overall fatigue, and morning fatigue) were relieved by 29% to 33% with sodium oxybate, compared with 6% to 10% relief with placebo (p < 0.005). Alpha intrusion, sleep latency, and rapid-eye-movement sleep were significantly decreased, while slow-wave (stage 3/4) sleep was significantly increased, compared with placebo (p < 0.005). Two of the 5 subjective sleep related variables were significantly different from placebo: morning alertness (improved by 18% with sodium oxybate, compared with 2% for placebo; p = 0.0033) and quality of sleep (improved by 33% and 10%, respectively; p = 0.0003). CONCLUSION: Sodium oxybate effectively reduced the symptoms of pain and fatigue in patients with FM, and dramatically reduced the sleep abnormalities (alpha intrusion and decreased slow-wave sleep) associated with the nonrestorative sleep characteristic of this disorder. PMID- 12734910 TI - Camptodactyly, arthropathy, coxa vara, and pericarditis syndrome among egyptians. AB - OBJECTIVE: To highlight the clinical, radiological, and pathological presentations of 10 Egyptian patients with camptodactyly, arthropathy, coxa vara, and pericarditis (CACP) syndrome. Methods. Ten cases underwent a full history, complete clinical examination, laboratory and radiological investigations (and magnetic resonance imaging, MRI, of knee joints); arthroscopic histopathological synovial studies were performed in 6 cases. Results. Camptodactyly and large joint arthropathies were present in all cases. The onset was at birth in 5 cases, and consanguinity was present in all cases. Laboratory investigations were normal in all cases (normal complete blood cell count, erythrocyte sedimentation rate, absent rheumatoid factor, and antinuclear antibody). Synovial fluid analyses were acellular in 3, hypocellular in 4, and moderately cellular in 2 cases. Histopathology revealed noninflammatory synovial hyperplasia in the 6 synovial biopsies obtained, with multinucleated giant cells in 4 of them. Plain hip radiology revealed short broad femoral neck and widening of joint space in all cases, with coxa vara in 9 cases. MRI of the knees showed rim-like enhancement of the lining of the fluid filled bursae in 7 cases, and homogenous enhancement pattern in 2 cases. No evident cartilage destruction existed in any case. CONCLUSION: Our cases represent a familial syndrome of noninflammatory arthropathies associated with camptodactyly and coxa vara. The complete picture of the syndrome may be related to disease duration, and MRI is a useful tool in diagnosis. Physicians should be aware of the syndrome, to avoid misdiagnosis with other pediatric connective tissue diseases. PMID- 12734911 TI - Juvenile spondyloarthritis and severe cardiac involvement in a female patient. AB - Heart involvement is a recognized complication in 10-20% of all adults with spondyloarthritis. Until now only 8 cases of cardiac involvement in juvenile spondyloarthritis (JS) have been reported, all male patients. We describe the first female patient with JS, in whom progressive cardiac involvement developed, and summarize the pediatric JS cases with cardiac involvement. PMID- 12734912 TI - Gastric antral vascular ectasia (watermelon stomach) in a patient with Sjogren's syndrome. AB - Gastric antral vascular ectasia (GAVE), a rare yet treatable cause of upper gastrointestinal bleeding, has been described in a variety of autoimmune diseases. We describe a patient who had typical Sjogren's syndrome and iron deficiency anemia requiring blood transfusion. An endoscopy showed characteristic findings of GAVE. After several fulguration therapies with argon-plasma coagulator, the mucosal lesions improved and her hemoglobin levels returned to normal. PMID- 12734913 TI - Interleukin 18 and hepatocyte growth factor in fulminant hepatic failure of adult onset Still's disease. AB - Adult onset Still's disease (AOSD) is a rheumatoid disorder characterized by polyarthritis, intermittent high fever, and salmon colored rashes. Liver dysfunction is usually mild and fulminant liver failure is rare. We describe a 20 year-old woman with AOSD and severe hepatic necrosis leading to hepatic failure requiring liver transplant. This severe liver disorder developed after decreases in fever, arthritis, and C-reactive protein. Interleukin 18 (IL-18), but not ferritin, increased in association with liver dysfunction. Hepatocyte growth factor (HGF) increased at the time of hepatic failure. IL-18 and HGF elevation may have contributed to this rare severe liver injury in AOSD. PMID- 12734915 TI - Ollier's disease with sella turcica involvement. PMID- 12734917 TI - OMERACT workshop: repair of structural damage in rheumatoid arthritis. AB - This article describes the process and results of a workshop aimed at reviewing data on repair of structural damage collected by the OMERACT Subcommittee on Healing of Erosions and at defining a priority list for the subsequent research agenda. PMID- 12734916 TI - Repair of erosions in rheumatoid arthritis does occur. Results from 2 studies by the OMERACT Subcommittee on Healing of Erosions. AB - The committee was charged with determining whether healing of erosions in rheumatoid arthritis (RA) occurs. Two exercises were performed: The first asked the committee members, as a panel of experts, to express agreement or disagreement with the presence of improvement and features of bone reaction to injury in images submitted by members as examples of healing. The second presented panel members with 28 pairs of serial images, 14 chosen to illustrate progression and 14 chosen to illustrate repair. Agreement was tested on 8 items: global judgment on which image in the pair was better, relative size of the erosion in the 2 images, judgment on which image was first, presence and extent of sclerosis, cortication, filling-in, remodeling, and reconstituting normal structure. Our results showed good agreement, among the 15 respondents, on global assessment of which image was better and which image showed the smaller erosion. Correct assignment of sequence was only slightly better than expected by chance (in 65% of the cases). Agreement was poor regarding the presence of morphologic features of bone repair. A majority of a panel of experts agreed on which 2nd images in a set of paired, serial images represented improvement and which showed progression based on global assessment of which was better and on size of erosion. Features of bone repair were not distinctive and did not enable the panel to deduce the correct sequence of the serial images. This study provides evidence that repair of bone damage in RA does occur, resulting in some degree of improvement, which was recognized by a majority of a panel of experts. PMID- 12734918 TI - MCID/Low Disease Activity State Workshop: low disease activity state in rheumatoid arthritis. AB - The MCID (minimal clinically important difference) module of OMERACT 5 developed a research agenda that led to the conclusion that a state of low disease activity for rheumatoid arthritis (RA) would need to be defined. To develop such a definition the various concepts and terminologies, the process for developing an operational definition, and the availability and design of longitudinal datasets for validation needed to be considered. This article describes the process of the MCID/Low Disease Activity State Workshop at OMERACT 6 to develop such a definition. PMID- 12734914 TI - Acute synovial rupture of the rheumatoid knee presenting as a pretibial mass: MRI appearance. PMID- 12734919 TI - Deriving an operational definition of low disease activity state in rheumatoid arthritis. AB - This article summarizes the process proposed to come to a definition of low disease activity in rheumatoid arthritis (RA). The purpose of this definition is to aid the interpretation of trial and longitudinal study results. A conceptual proposal is "a disease activity state that is deemed a useful treatment target by both physicians and patients." An operant definition can be derived by judgmental (opinion-based) or statistical (data-based) approaches, but the first seems more appropriate. Once a few candidate definitions have been selected, their usefulness and prognostic validity can be tested in longitudinal datasets. PMID- 12734920 TI - MCID/Low Disease Activity State Workshop: summary, recommendations, and research agenda. AB - The OMERACT 6 Minimal Clinically Important Difference/Low Disease Activity Workshop was organized with the aim of meeting the many challenges that exist in determining a low disease activity in rheumatoid arthritis (RA). This article presents an overview of that workshop, including results of the voting, a summary of associated discussions, recommendations, and a proposed research agenda. PMID- 12734921 TI - Sex differences in giant cell arteritis. PMID- 12734922 TI - HLA-B27 and free HLA class I heavy chains in ankylosing spondylitis. PMID- 12734923 TI - Cyclophosphamide pulse regimen in the treatment of alveolitis in systemic sclerosis. PMID- 12734925 TI - Neck and upper limb pain. PMID- 12734924 TI - Consistency for childhood arthritis. PMID- 12734926 TI - Fluvoxamine therapy for fibromyalgia. PMID- 12734927 TI - Salicin and treatment of rheumatic diseases. PMID- 12734928 TI - [Effect of HIV-1 infection on apoptosis of CD4+ T lymphocytes mediated by Fas]. AB - OBJECTIVE: To study the pathogenic role of Fas/CD95 in HIV-1 infection subjects, and to investigate the effects of HIV on plasma levels of sFas and the expression of CD95 on different CD4(+) T lymphocyte subpopulations. METHODS: Four-color flow cytometry was used to determine the expression of CD95, CD45RO, CD45RA on CD4(+ )T lymphocyte in peripheral blood from HIV-1 infection subjects and serum Fas levels were quantified by enzyme-linked immunosorbent assay (ELISA). RESULT: Compared with healthy controls, serum Fas levels were significantly increased (P<0.05) in HIV group and positively correlated with the disease progress. The expression of CD95 on naive T-lymphocyte subsets was increased whereas that on memory T-lymphocyte subsets was decreased. CONCLUSION: Fas plays an important role in the deletion of CD4(+) T-lymphocyte during HIV-1 infection. Further understanding of the relationship between Fas/CD95 and CD45RO/CD45RA may help to predict the progression of the disease and the clinical outcome. PMID- 12734929 TI - [Evaluation of serum interleukin-18 and interleukin-10 in patients with HIV-1 and hepatitis viruses co-infected subjects]. AB - OBJECTIVE: To study the significance of cytokines in patients with HIV and hepatitis viruses co-infection. METHODS: Serum levels of IL-18 and IL-10 were measured by enzyme-linked immunosorbent assay (ELISA). HIV-RNA levels were measured in EDTA plasma by quantitative reverse polymerase chain reaction (PCR). CD4(+) lymphocyte counts were determined by four-color Flow cytometry (FCM). RESULTS: The levels of IL-18 were significantly higher in HIV-infected persons compared with those in controls (P<0.05). With HIV disease progression, IL-18 levels increased while Il-10 levels decreased. HCV patients showed lower levels of IL-18 and IL-10 than those of the co-infection group. CONCLUSION: Univariate analyses shows significant co-variables IL-10 in co-infection. Up-regulating IL 18 activity and/or down-regulating IL-10 may be a potential therapy to patients with HIV and hepatitis viruses co-infection. PMID- 12734931 TI - [Immune activation in AIDS related Kaposi's sarcoma]. AB - OBJECTIVE: To study the pathogenesis role of immune system activation in AIDS related Kaposi's sarcoma(AIDS-KS). METHODS: The serum levels of sFas, beta 2 microglobin, IL-10, IL-16, IL-18, IL-6 and sIL-4R were detected by ELISA in 8 AIDS-KS patients, 28 patients with HIV infection but without Kaposi's sarcoma(HIV NKS) and 16 normal controls. The lymphocyte and their subsets, CD38(+) CD8, HLA DR(+)CD8 in the peripheral blood mononuclear cell (PBMCs) in 12 AIDS-KS and 32 HIV-NKS were detected by flow cytometer. RESULTS: Beta 2-MG and sIL-4R in HIV-NKS were significantly higher than those in normal controls(P<0.05), IL-16 in HIV-NKS was significantly lower than that in controls(P<0.05). IL-18 was higher in both AIDS-KS and HIV-NKS compared with normal controls. In AIDS-KS, CD3, CD4, CD8, NK and HLA-DR(+)CD8 were lower than those in HIV-NKS whereas CD19 and CD38(+)CD8 were higher than those in HIV-NKS. But the difference was not statistically(P<0.05). CONCLUSION: Although both AIDS-KS and HIV-NKS demonstrate some activation of immune system, there appears to be no significant difference between immune responses in KS and NKS patients. These data suggest that the activation of the immune system is unlikely to contribute significantly to the pathogenesis of AIDS-KS. PMID- 12734930 TI - [Expression of co-receptor on different T lymphocytes subpopulations after effective HAART]. AB - OBJECTIVE: To measure CCR5 and CXCR4 chemokine receptor expression on CD4 and CD8 T cells in HIV-1 infection and to relate levels to the distribution of CD45RO memory and CD45RA-naive subsets after effective HAART. METHODS: Four-color cytofluorometry with appropriate conjugated monoclonal antibodies (mAbs) was performed to define CD45RA and CD45RO subsets of CD4 and CD8 T cells and measure the expression of CCR5, CXCR4 in blood from 43 received HAART patients and 5 non treated HIV and 13 healthy controls. RESULTS: The levels of CCR5 and CXCR4 on CD4 and CD8 T cells and their CD45RO/CD45RA subsets in HIV-1-infected patients had not any statistical significance than that on control subjects and effective HAART could adjust the expression on T cells. CONCLUSION: CXCR4/CCR5 plays an important role in the progress of HIV-1 infection. The most favorable condition for treatment should be initiated before stage B. PMID- 12734932 TI - [Drug resistance of HIV-infected patients after the failure of highly active antiretroviral treatment]. AB - OBJECTIVE: To find out the mechanism of drug resistance by detecting the mutations of HIV RNA in patients who failed in the anti-HIV therapy, to direct the clinical use of anti-HIV drugs and to complement the existing drug resistant database. METHODS: HIV RNA and DNA were extracted from the plasma of 10 HIV infected patients who developed drug resistance in the Clinic of AIDS, Ruhr University, Bochum, Germany. Then HIV-RNA was amplified in the reverse transcriptase (RT) and protease regions by polymerase chain reaction (PCR). After purified, the PCR products was sequenced. The acquired sequences were compared with the international standard strain HXB2CG and the resistant database of Stanford University. RESULTS: Some mutations were found to cause the corresponding resistance to certain drugs and were consistent with the clinical results. Some mutations existed in some patients, such as V179I in RT and K20T, K20I in protease, which hadn't been reported in the resistant database of Stanford University yet. CONCLUSION: Patients who fail in HAART have different mutations in RT and protease regions. Mutations such as V179I in RT and K20T, K20I etc in protease may be related to drug resistance. PMID- 12734933 TI - [Influence of HGV super-infected with HIV or HCV on the virus replication]. AB - OBJECTIVE: To realize human immunodeficiency virus(HIV) and hepatitis C virus(HCV) super-infected with hepatitis G virus(HGV or GBV/C) and to probe into the mechanism of these virus infection in the body. METHODS: HIV and HCV load were tested by the quantitated RT-PCR in the HIV or HCV infected plasma samples respectively and the HGV RNA was detected in all of the samples. Then some of the HGV positive were sequenced. RESULTS: 123 of 317 HIV patients were positive for HGV, the positive rate was 38.8%. Among the 91 HCV patients, 19 were positive for HGV. The positive rate is 20.9% which was less than that of HIV patients. HIV load of the patients super-infected with HGV was less than that of those without HGV[(1.8+/-0.6)x10 copies/ml compared with (1.9+/-1.1)x10(2)copies/ml]; while HGV and HCV super-infection did not influence the HCV RNA load significantly [(1.5+/ 0.6)x10(4) copies/ml compared with (5.4+/-1.8)x10(4)copies/ml]. The HGV sequences from HIV or HCV patients were compared and showed no difference markedly. CONCLUSION: The rate of the HIV and HGV super-infection is higher than that of HCV. HGV may inhibit HIV reproduction in the body while superinfection. PMID- 12734934 TI - [DNAzymes in vitro inhibit the expression of hepatitis B virus genes]. AB - OBJECTIVE: To evaluate the inhibition effects of DNAzymes specific to Hepatitis B Virus(HBV) s gene and e gene on the expressions of Hepatitis B surface antigen(HBsAg) and Hepatitis B e antigen(HBeAg). METHODS: DNAzymes DrzBS and DrzBC specific to HBV s gene ORF A157UG and e gene ORF A1816UG, respectively, were designed and synthesized. The inhibition effects of DrzBS or DrzBC on the expressions of HBV s and e genes were observed in 2.2.15 cells. RESULTS: The expression of HBV s or e genes was dramatically depressed after 2.2.15 cells treated by DrzBS or DrzBC. The concentration for effective inhibition was within 0.1-2.5 micromol/L and the inhibition showed a dose dependence within that concentration range. The maximum inhibition was 94.2% and 91.8% for DrzBS and DrzBC, respectively. The inhibition was maintained for 72 hours. The efficiency of inhibiting HbsAg, HbeAg in 2.2.15 cells by DrzBS, DrzBC was higher than that by antisense oligonucleotides for the same target genes. The concentrations for effective inhibition of the DNAzymes were at least 10-fold lower compared with antisense oligonucleotides. Neither inhibition on the replication of HBV DNA nor toxicity to 2.2.15 cells was observed. CONCLUSION: DrzBS and DrzBC can highly block the expressions of HBV s gene and e gene in 2.2.15 HBV cell model and are proved a specific and effective anti-HBV gene therapeutic means. PMID- 12734935 TI - [Stereoselective determination of propranolol enantiomer in transgenic cell lines expressing human cytochrome P450]. AB - OBJECTIVE: To establish a chiro chromatography for studying the stereoselective metabolism of propranolol (PL) in S(9) incubates prepared from transgenic cell lines expressing human cytochrome P450. METHODS: The concentration of each enantiomer in S(9) incubates was determined through precolumn derivatization with GITC, followed by RP-HPLC assay using S-(+)-propafenone as internal standard. RESULTS: Baseline separations among the diastereomers of S(-)-P, internal standard and R(+)-PL were achieved on Shimpack CLC C(18)ODS column, with UV detection and methanol:water:glacial acetic acid (67/33/0.05,v/v/v) as mobile phase. The assay was simple, accurate, precise and specific. The linear range was from 5 to 500 micromol/L for each enantiomer. The limit of quantitation (LOQ) for the method was 5 micromol/L for the S(-)-and R(+)-PL, respectively (n=5, RSD<10%). The analytical method afforded average recoveries of 98.7 and 98.1% for S(-)- and R(+)-PL, respectively. The reproducibility of the assay was good (RSD<10%). The time-dependent studies showed that PL had the stereoselectivity of S-(-)-isomer in metabolism via CYP2C18 and the stereoselectivity of R-(+)-isomer in metabolism via CYP2C9. CONCLUSION: The method allows to study of stereoselective metabolism of PL in vitro. PMID- 12734936 TI - [Improved alternative electro-stimulus Y-maze for evaluating the spatial memory of rats]. AB - OBJECTIVE: To establish an effective model for evaluating the spatial memory of rats. METHODS: The adjacent left or right arm in Y-maze was defined as an initial arm or an objective one alternatively, and electro-stimulus to vola was used as a reinforcement. Meanwhile, the reliability of the model was validated by measuring the memory deficits induced by intraperitoneal (i.p.) injection of scopolamine (0.5 mg/kg) or bilaterally intrahippocampal (i.h.) microinjection of MK-801(0.2 microg/site). RESULTS: The normal, sham-operated or saline-treated rats learned the left-right alternative memory model rapidly while injection of either scopolamine or MK-801 caused spatial memory deficits in Y-maze, including deficits in working and reference memory. Moreover,there was no appreciable difference in electricity intensity between the treated group and the control group. CONCLUSION: The alternative Y-maze with electro-stimulus is a comparatively objective model which can evaluate the spatial memory in rats effectively.T PMID- 12734937 TI - [Effect of PP60c-Src on angiotensin II-induced signal transduction in rat vascular smooth muscle cells]. AB - OBJECTIVE: To further clarify the mechanism of Ang II-induced intracellular signal transduction in vascular smooth muscle cells(VSMCs) proliferation by observing the effect of c-Src on Ang II-mediated MAPK activation and c-fos protein expressions in rat VSMCs. METHODS: Aortic VSMCs from SD rats were cultured primarily and subcultured, which were transfected with anti-sense c-Src oligodeoxynucleotides(ODNs) wrapped with lipofectin to inhibit c-Src activity and protein production. Untransfected VSMCs were used as control, to observe the role of Ang II stimulation in MAPK activation and c-fos protein expression in VSMC. Protein immunoprecipitation and kinase phosphorylation were employed to measure c Src kinase activity; MAPK kinase activity was assessed by the phosphorylation rate of the substrate MBP(Myelin Basic Protein); Western blot was used to assess the protein expression of c-Src and c-fos. RESULTS: c-Src protein expressions in VSMC, which were transfected with different concentrations of anti-sense c-Src ODNs, were significantly decreased in a negative dose-effect manner (0.2 microm, 0.5 microm, 1.0 microm and 2.0 microm were 68.2%, 34.7%, 30.3% and 15.8% respectively compared with control). c-Src kinase activity was also obviously inhibited. Following stimulation of Ang II on VSMC transfected with anti-sense c Src ODNs, the increase of c-Src activity was only 8.7% of control,the activity of MAPK only 1.6% compared with control, and the increase in c-fos protein expression 30.3% as control. CONCLUSION: Ang II can induce c-Src activation and intracellular signal transduction in VSMC which depend on c-Src activation, indicating that c-Src is a pivotal signal factor in VSMC proliferation. PMID- 12734938 TI - [Developmental expression and regulation of divalent metal transporter 1 in rat heart]. AB - OBJECTIVE: To investigate the expression of divalent metal transporter 1 (DMT1) mRNA in male Sprague-Dawley rat heart of different ages and the expression of DMT1 regulated by dietary iron. METHODS: Reverse transcriptase (RT)-PCR and Western blot were used in this study. RESULTS: (1)Two isoforms of DMT1 mRNA [with and without iron-responsive element (IRE)] were both detected in rat heart, which were correlated with heart iron content. During development, both of two isoforms of DMT1 mRNA expression were the lowest at the age of PND 7, and increased at PND 21, 63 to 196. (2) After fed with a high iron diet or low iron diet for 6 weeks, the rats developed iron overload or iron deficiency respectively. No significant differences in DMT1 mRNA expression were detected among iron overload, iron deficiency and control rats. By using Western blot analysis, a 21% and 40% reduction in DMT1 protein non IRE form and IRE form respectively were found in iron overload rat (P<0.01, compared with control). Increases (26% approximate, equals 28%) in the levels of two isoforms of DMT1 protein were also observed in iron deficient rat (P<0.01, compared with control). CONCLUSION: The level of DMT1 mRNA expression in heart is age dependent;the two isoforms of DMT1 protein may be both regulated by iron on the posttranscriptional mechanism. PMID- 12734939 TI - [Isolation, culturation and adipogenisis committed differentiation of adult human mesenchymal stem cell]. AB - OBJECTIVE: To isolate MSCs from adult human bone marrow cells and to induce them into adipocytes. METHODS: MSCs were isolated from adult human bone marrow aspirated by Percoll and expanded in L-DMEM. The surface antigen of MSCs, CD14, CD34, CD45, CD44, VLA-1, HLA-DR and cell cycle were analysed on a FACScan flow cytometer. MSCs were cultured in adipogenisis inducing medium including insulin, 1-methyl-3-isobutylxanthine, indomethine and dexamethasone for 7 days and stained with Oil Red O. RESULTS: MSCs grew as adherent cells and expanded more than 10 passages. They were positive for CD44 and negative for CD14, CD34, CD45, HLA-DR. The expression of VLA-1 was weak. After 7 days of adipocyte inducing, about 85%of the cells displayed accumulation of lipid vacuoles, as detected by Red Oil O. CONCLUSION: MSCs isolated and cultured from adult human bone marrow can be induced to adipogenisis committed differentiation. PMID- 12734941 TI - [Approach of influence factors on infectious complications in patients with primary nephrotic syndrome]. AB - OBJECTIVE: To explore the influence factors on infectious complications in the patients with primary nephrotic syndrome in the course of using glucocorticoid and immune inhibitors. METHODS: 571 hospitalized and some outpatients with primary nephrotic syndrome were retrospectively analysed from January,1992 to June,2001 to explore the relationship between the infectious complication and the dose of glucocorticoid,or the curative course of glucocorticoid, or whether glucocorticoid was united with other immune inhibitors,or the course of disease. RESULTS: The infectious incidence increased gradually and the degree became more serious along with the increase of the glucocorticoid dose and the prolonging of the curative course. To the patients who failed in the treatment of prednisone(dose > or = 1 mg/(kg.d), curative course 1 month or so), the dose of the prednisone was reduced to 0.5 mg/(kg.d) or so and multiglycosides tripterygii wilfordii tablets or cyclophosphamide united. The infectious complications was decreased and the degree was alleviated obviously for the patients treated with this method for more than 2 months compared with those who only used prednisone(dose > or = 1 mg/(kg.d), curative course >3 month). There was no obvious difference in the infectious incidence among the groups of different course of disease before using glucocorticoid. CONCLUSION: Glucocorticoid and immune inhibitors must be reasonably applied in order to reduce and alleviate infectious complications for treating primary nephrotic syndromes. PMID- 12734940 TI - [Detection of thrombomodulin in both plasma and tissue extracts of cancer patients and its clinical significance]. AB - OBJECTIVE: To study the changes of thrombomodulin(TM) in both plasma and tissue extracts of cancer patients for evaluating its clinical significance. METHODS: Plasma TM levels were measured by enzyme-linked immunosorbent assay(ELISA) in plasma of 188 cancer patients and in 24 cancer tissue extracts including their adjacent normal tissue. RESULTS: The plasma TM levels both in cancer patients and in metastasis patients were significantly higher than that in controls [(33.47+/ 14.25) microg/L/ (41.68 +/-16.96) micro/L, compared with (20.40+/-7.22) microg/L, P<0.01]. The plasma TM levels in cancer patients after operation decreased obviously [(18.45+/-9.96) microg/L, compared with (28.29+/-11.74) microg/L,P<0.01]. Whereas, the plasma TM levels in patients with recurrence and metastasis after operation increased obviously [(34.50+/-12.57 micro/L]. The plasma TM levels in metastasis of lung cancers, gastric cancers and pancreatic cancers were significantly higher than that in non-metastasis (P<0.05 approximate, equals 0.01) respectively, but no significant differences were found between controls and non-metastasis cancers including gastric cancers, pancreatic cancers, nasopharyngeal cancers, large intestine cancers and laryngeal cancers (P>0.05). The TM levels in cancer tissue extracts were significantly lower than that in their adjacent normal tissue extracts [(647.71+/-317.51)) microg/L,compared with (1455.63+/-772.22) microg/L,P<0.01]. On the contrary, the plasma TM levels in these cancers were higher than that in controls. CONCLUSION: The rise of plasma TM levels in cancer patients is associated with metastasis and diffusion of cancers. The TM levels can be used as an sensitive index for judging progression and metastasis of cancers. PMID- 12734942 TI - [Postoperative hemodynamic parameter follow-up in children with severe pulmonary resistance hypertension due to ventricular septal detect]. AB - OBJECTIVE: To study the relationship between the age at operation and prognosis in children with severe pulmonary hypertension (PH) due to ventricular septal defect (VSD). METHODS: Forty children with severe PH (increased total pulmonary circulation resistance)due to VSD were divided into two groups based on age at operation(Group I aged less than 2 years and group II more than 2 years). The hemodynamic parameter follow-up was measured by cardiac catheterization at presurgery, one week after surgery and 5-7 years postoperatively. RESULTS: The ratio of pulmonary arterial pressure and systemic arterial pressure (pp/ps),pulmonary resistance and systemic resistance (R(p)/R(s)), and pulmonary vascular resistance (PVR) and small pulmonary arterial resistance (PAR) were significantly different in two groups (P<0.01). During follow-up in the group less than 2 years, all the hemodynamic parameters were at normal level, while in the group more than 2 years, only p(p)/p(s) and R(p)/R(s) were close to normal level. The pulmonary arterial resistance was still abnormal. CONCLUSION: An early operation may be the only way to gain optimal long term result of surgery and decrease the incidence of pulmonary vascular disease in children with PH due to VSD. PMID- 12734944 TI - [Vascular complications after orthotopic liver transplantation]. PMID- 12734943 TI - [Prediction of human skin permeability of drugs in vivo with artificial neural network]. AB - OBJECTIVE: To predict in vivo human skin permeability of drugs. METHODS: Appropriate BP(Back-propagation) neural network to predict human skin permeation ratios of drugs (R, absorbed/unabsorbed) was established. The octanol water partition coefficients (logP), molecular volumes (V), hydrogen bond acidities (sigma alpha 2(H)) and hydrogen bond basidities (sigma beta 2(H)) were selected as the neural units of input layer, and logR values were selected as the neural units of output layer. RESULTS: The calculated logR values of 17 drugs were in good accordance with their observed values. CONCLUSION: BP neural network can be used to predict in vivo human skin permeability of drugs. PMID- 12734945 TI - [Analysis and treatment of the recurrent retinal detachment after silicone oil injection]. PMID- 12734946 TI - [Genetic and epigenetic effects of DNA methylation in cancer]. PMID- 12734947 TI - [Anatomical principles of the coronary artery system]. PMID- 12734948 TI - [Basic structure of microcirculatory system in the human heart]. PMID- 12734949 TI - [Coronary hemodynamics]. PMID- 12734950 TI - [Basic regulatory mechanisms of coronary circulation]. PMID- 12734952 TI - [Cardiac energetics and coronary circulation]. PMID- 12734951 TI - [Coronary flow reserve]. PMID- 12734954 TI - [Development and progression of coronary atherosclerosis]. PMID- 12734953 TI - [Coronary circulation and cardiac function]. PMID- 12734955 TI - [Coronary arteriosclerosis and lipid metabolism (hyperlipemia)]. PMID- 12734956 TI - [Coronary atherosclerosis and vascular endothelial dysfunction]. PMID- 12734958 TI - [Coronary artery sclerosis and vascular remodeling]. PMID- 12734957 TI - [Vascular smooth muscle cells in coronary heart disease]. PMID- 12734959 TI - [Pathophysiology of coronary vasospasm]. PMID- 12734960 TI - [Pathomorphology of thrombus formation and its progress in coronary artery]. PMID- 12734961 TI - [Coronary thrombosis and coagulation/fibrinolytic system]. PMID- 12734962 TI - [Nonatherosclerotic coronary artery disease]. PMID- 12734964 TI - [Stunned myocardium and hibernating myocardium: conception, definition and pathophysiology]. PMID- 12734963 TI - [Mechanical property of ischemic myocardium]. PMID- 12734965 TI - [No reflow phenomenon]. PMID- 12734966 TI - [The trend of research on the mechanism of reperfusion injury]. PMID- 12734967 TI - [Anti-ischemic tolerance and preconditioning]. PMID- 12734968 TI - [Cell regeneration in infarcted myocardium]. PMID- 12734969 TI - [Neovasculation in ischemic heart disease]. PMID- 12734970 TI - [Electrocardiography and vectorcardiography]. PMID- 12734971 TI - [Holter electrocardiogram]. PMID- 12734972 TI - [Exercise testing]. PMID- 12734974 TI - [Signal averaged electrocardiogram]. PMID- 12734973 TI - [Body surface potential mapping]. PMID- 12734975 TI - [Clinical cardiac electrophysiological study]. PMID- 12734976 TI - [Quantitative coronary angiography: QCA]. PMID- 12734977 TI - [Arteriography with synchrotron radiation--principle, characteristics and problems]. PMID- 12734978 TI - [The current progress and applications of echocardiography for diagnosis of ischemic heart disease]. PMID- 12734979 TI - [Noninvasive echocardiographic examination]. PMID- 12734980 TI - [Intravascular ultrasound (IVUS)]. PMID- 12734981 TI - [Intravascular ultrasonic Doppler systems]. PMID- 12734982 TI - [Fundamental theory and recent advances of electron beam CT]. PMID- 12734983 TI - [Multislice . helical CT, 3DCT(coronary angiography)]. PMID- 12734984 TI - [MRI of ischemic heart disease]. PMID- 12734986 TI - [Basic theory of fast scan cine MRI]. PMID- 12734985 TI - [Gd-DTPA enhanced MRI]. PMID- 12734987 TI - [MRI tagging]. PMID- 12734988 TI - [MR coronary angiography: a review of fundamental theory and recent advances]. PMID- 12734989 TI - [Cardiac magnetic resonance spectroscopy and chemical shift imaging]. PMID- 12734991 TI - [Myocardial perfusion imaging]. PMID- 12734990 TI - [Advance in nuclear cardiology and radiopharmaceuticals]. PMID- 12734992 TI - [99mTc-PYP scintigraphy: its usefulness and pitfalls]. PMID- 12734993 TI - [Myocardial fatty acid imaging with I-123 BMIPP]. PMID- 12734995 TI - [Radionuclide ventriculography and gated blood-pool SPECT in coronary artery disease]. PMID- 12734994 TI - [I-123 metaiodobenzylguanidine myocardial images for assessment of coronary artery disease]. PMID- 12734996 TI - [201Tl, 99mTc-PYP dual SPECT]. PMID- 12734997 TI - [201Tl, 123I-BMIPP dual SPECT]. PMID- 12734998 TI - [Exercise stress thallium-201 myocardial scintigraphy]. PMID- 12734999 TI - [Dipyridamole stress thallium-201 myocardial scintigraphy]. PMID- 12735001 TI - [Dobutamine stress myocardial single photon emission computed tomography]. PMID- 12735000 TI - [ATP stress thallium--201 myocardial scintigraphy]. PMID- 12735002 TI - [Positron emission tomography (PET)]. PMID- 12735003 TI - [Progress in coronary angioscope]. PMID- 12735004 TI - [Angioscopic findings of coronary atherosclerosis]. PMID- 12735005 TI - [Angioscopic assessment of the characteristic of coronary-arteries thrombi]. PMID- 12735006 TI - [Electro anatomical analysis (NOGA, CARTO system)]. PMID- 12735007 TI - [Cardiopulmonary exercise testing]. PMID- 12735008 TI - [Magnetocardiograms]. PMID- 12735010 TI - [Indication and contraindication for intracoronary thrombolysis]. PMID- 12735009 TI - [Biochemical diagnosis, molecular genetics method]. PMID- 12735011 TI - [Variety and selection of thrombolytic agents]. PMID- 12735012 TI - [Thrombolytic agents, urokinase, tissue-type plasminogen activator and pro urokinase--their routes of administration and dosage: comparison between intracoronary thrombolysis and intravenous coronary thrombolysis]. PMID- 12735013 TI - [Intravenous coronary thrombolysis on evidence based reperfusion therapy for acute myocardial infarction]. PMID- 12735014 TI - [Mutant t-PA compared with native t-PA in acute myocardial infarction]. PMID- 12735015 TI - [GP IIb/IIIa receptor prevention medicine]. PMID- 12735016 TI - [Clinical markers of reperfusion: chest pain and electrocardiogram]. PMID- 12735017 TI - [Image diagnosis]. PMID- 12735018 TI - [Detection of coronary reperfusion using biochemical markers]. PMID- 12735019 TI - [Residual stenosis after intracoronary thrombolysis and usefulness of delayed angioplasty]. PMID- 12735020 TI - [Anti thrombotic therapy after coronary thrombolysis]. PMID- 12735021 TI - [PCI: results of nationwide survey in Japan]. PMID- 12735023 TI - [Elective PTCA for angina pectoris]. PMID- 12735022 TI - [Indications for percutaneous coronary intervention]. PMID- 12735024 TI - [Percutaneous transluminal coronary angioplasty (PTCA) for acute myocardial infarction]. PMID- 12735025 TI - [The incidence and mechanism of restenosis after PTCA]. PMID- 12735026 TI - [Complication of percutaneous coronary intervention--coronary dissection and acute closure]. PMID- 12735027 TI - [Prevention of restenosis after PTCA and patient's QOL]. PMID- 12735028 TI - [Percutaneous coronary intervention: Current status of the development of new devices and the perspective of their future]. PMID- 12735029 TI - [Operative transluminal coronary angioplasty (OTCA)]. PMID- 12735030 TI - [Laser coronary angioplasty]. PMID- 12735031 TI - [Transmyocardial laser revascularization (TMLR), percutaneous myocardial laser revascularization (PMLR)]. PMID- 12735032 TI - [CABG: the world trends and the Japanese realities]. PMID- 12735033 TI - [Indication for coronary artery bypass grafting]. PMID- 12735034 TI - [Choice & variations of CABG grafts]. PMID- 12735035 TI - [Long-term prognosis of coronary artery bypass surgery in Japan]. PMID- 12735036 TI - [Evaluation of coronary artery bypass graft patency]. PMID- 12735037 TI - [Investigation into the cause of re-stenosis after coronary bypass surgery]. PMID- 12735039 TI - [Significance of collateral circulation on CABG]. PMID- 12735038 TI - [Coronary artery bypass : Vein graft lesion]. PMID- 12735040 TI - [Indications and results of emergency coronary artery bypass grafting in unstable angina]. PMID- 12735041 TI - [Minimally invasive CABG MIDCAB/OPCAB]. PMID- 12735043 TI - [Cost analysis of percutaneous coronary intervention and coronary artery bypass grafting in Japan and USA]. PMID- 12735042 TI - [Prevention of postangioplasty restenosis]. PMID- 12735044 TI - [Thrombolytic (fibrinolytic) agents for treatment of ischemic heart disease]. PMID- 12735045 TI - [Anticoagulant, antiplatelet agents]. PMID- 12735046 TI - [Antianginal drugs]. PMID- 12735047 TI - [Pharmacological treatment of heart failure]. PMID- 12735048 TI - [The protection of myocardial ischemia--reperfusion injury]. PMID- 12735049 TI - [Treatment of high blood cholesterol in patients with coronary heart disease]. PMID- 12735050 TI - [Gene therapy for restenosis after angioplasty]. PMID- 12735052 TI - [Therapeutic angiogenesis for ischemic heart disease by transplantation of bone marrow cells]. PMID- 12735051 TI - [Therapeutic angiogenesis by angiogenic growth factor]. PMID- 12735053 TI - [Transmyocardial laser revascularization (TMR)]. PMID- 12735054 TI - [Cardiomyocyte regenerate therapy]. PMID- 12735055 TI - [Tissue engineering for ischemic heart disease]. PMID- 12735056 TI - [Epidemiological statistics of ischemic heart disease in Japan]. PMID- 12735058 TI - [Coronary risk factors in a rural community and an urban community]. PMID- 12735057 TI - [Trends in risk factors for ischemic heart disease among Japanese]. PMID- 12735059 TI - [Dyslipoproteinemia as a coronary risk factor]. PMID- 12735060 TI - [Effect of coronary risk factors on characteristics of coronary atherosclerosis]. PMID- 12735061 TI - [Risk factors for ischemic heart disease in young adults]. PMID- 12735062 TI - [Psychosocial factors and coronary heart disease]. PMID- 12735063 TI - [Type A behavior pattern as a coronary risk factor]. PMID- 12735064 TI - [Prevention program for ischemic heart disease]. PMID- 12735065 TI - [New biochemical and genetic risk factors for coronary artery diseases]. PMID- 12735066 TI - Nurse educator shortage. New program approach. AB - The nursing shortage is here and now. Nursing faculty are againg and retiring, enhancing the current and projected faculty shortage. A skilled clinician is not automatically a skilled educator; the different roles involve different skill sets. The Nurse Educator Certificate for advanced practice clinicians offers one answer to the looming shortages. The new web-based educator certificate provides flexibility for students preparing to meet the rapidly changing healthcare needs of our population. Students benefit from an opportunity to improve their educator skill set in a convenient mode from their hoem communities. PMID- 12735067 TI - Fast facts about the Nurse Reinvestment Act to address the nursing shortage. PMID- 12735068 TI - Kansas nursing wage survey. PMID- 12735069 TI - Hospitals for a healthy environment. Partnership brings national healthcare P2 program to Kansas. PMID- 12735071 TI - "Why did he do it?" Confronting issues of suicide and bereavement. PMID- 12735072 TI - Suicide ideation among parents bereaved by the violent deaths of their children. AB - Suicidal behaviors of parents bereaved by a child's suicide have received considerable attention by researchers, but deaths by other violent causes have not. We observed 175 bereaved parents for five years following three types of violent death: accidents, homicides, and suicides. The results showed that the incidence of suicidal ideation (SI) among the study parents was 13% (n = 34) over the 5 years and 9% (n = 24) at the initial data collection four months after the death of an adolescent or young adult child. Comparisons of study parents grouped by the presence or absence of SI showed that after corrections were made for the number of t-tests conducted, statistically significant differences on three of four outcome variables remained (mental distress, depression, and posttraumatic stress disorder [PTSD], but not on acceptance of the child's death). The groups also differed significantly on four of seven mediating variables examined. The hypothesis that parents whose children died by suicide would report the highest incidence of suicidal ideation was not supported. Regression analyses showed that SI was a significant predictor of depression one year, but not five years, after the violent death of a child. The hypothesis that SI would predict both depression and PTSD one year postdeath was not supported. Clinical and policy recommendations are offered. PMID- 12735073 TI - Suicide and schizophrenia: a review of literature for the decade (1990-1999) and implications for mental health nursing. AB - This paper presents an overview of recent evidence on general and specific risk factors for suicide in patients with schizophrenia. The authors highlight the significant factors contributing to the likelihood of suicide in patients diagnosed with schizophrenia. This information will enhance the delivery of nursing care to these patients in all health care settings. A review of literature was conducted by two methods of investigation: Medline and CINAHL search and a manual search through articles from 1990 to 1999. The lifetime risk of committing suicide is estimated at about 9-13% of persons with schizophrenia, and it is 20 to 50 times higher than that in the general population. Young white males diagnosed with schizophrenia who are depressed, unmarried, unemployed, socially isolated, and functionally impaired and who lack external support are the most vulnerable in the early stages of schizophrenic illness. Findings can be instrumental in identifying and treating patients who are most vulnerable and in making psychiatric nurses aware of the scenarios and critical stages of the disease process when suicide is most likely to occur. PMID- 12735074 TI - Nurse-assisted suicide: not an answer in end-of-life care. PMID- 12735075 TI - Killing while caring: the nurses of Hadamar. AB - This research was conducted to examine the participation of nurses in the euthanasia program as it existed in the time of National Socialism in Germany and to describe the participation of nurses at one of the six killing centers, Hadamar, located near Frankfurt, Germany. Over 10,000 mentally and physically handicapped patients were killed by nurses as a part of the euthanasia programs, with over 100,000 being killed in all. Factors influencing the nurses' willingness to kill are described and include the socialization of the German people toward euthanasia as well as ideological commitment, economic factors, and putative duress. PMID- 12735076 TI - Client narratives: a theoretical perspective. AB - The role of subjective client narratives in health care represents a clinical and therapeutic tool, useful in complementing objective, scientific data. Of particular interest to mental health practitioners is the role narratives play as a therapeutic tool to guide clinical practice. This paper lays a foundation for understanding the importance of narrative in the psychotherapeutic process. It provides a brief overview of narrative theory and methods of structural analysis in order to provide a theoretical approach that can be utilized by nurses to address clients' needs. PMID- 12735078 TI - The influence of social support and health on everyday problem solving in adult African Americans. AB - The purpose of this study was to examine the relationship between social support, health status, and everyday problem solving in African Americans. The sample included subjects recruited from Baltimore, Maryland. The sample consisted of 249 community-dwelling African American adults, 32% of whom were male, with a sample mean age of 67.8 years (SD = 8.47 years). Variables included: Everyday Problem Solving Test (EPT), social support given and received, physical limitations, counts of chronic illness, smoking, and demographic information. Using stepwise regression, age, education, physical limitations, and social support given were found to be significant predictors of performance on the EPT. Further analysis found support for a partial mediating effect of physical limitations on the relationship between social support and everyday problem solving. The results indicate that there may be differences in the cognitive abilities of those actively involved in social activities. PMID- 12735077 TI - The use of narrative data to inform the psychotherapeutic group process with suicide survivors. AB - While bereavement is considered by many to be among one of the most stressful life events, it becomes even more distressing when it is related to the suicide of a loved one. A synopsis of psychosocial outcomes of suicide survivor bereavement is presented along with an overview of group interventions designed to help these survivors cope with their grief. The structure of an ongoing eight week bereavement support group is described to lay a foundation for the application of narrative theory within the group process. Using narrative theory and structural analysis, the discourse of group members is presented and various themes are discussed in an effort to contribute to the task of developing effective psychotherapeutic group interventions for survivors of suicide. PMID- 12735079 TI - Implicit and explicit measures of memory for perceptual information in young adults, healthy older adults, and patients with Alzheimer's disease. AB - In this study, we examined how implicit and explicit memory for perceptual information (modality and voice) and lexical information varied across three subject groups: healthy young adults, healthy older adults, and age-matched older adults with dementia of the Alzheimer's type (DAT). These groups exhibited cross modality (abstract) priming of the same magnitude. However, young adults produced greater modality- and voice-specific priming than the other two groups, whose performance was equivalent, suggesting that aging, but not DAT, reduced form specific priming. Young adults demonstrated better recognition memory than healthy older adults, who in turn exhibited better recognition memory than older adults with DAT. In young adults, recognition memory was also sensitive to perceptual information. These findings indicate that aging can affect implicit memory for perceptual information, whereas DAT magnifies the effect of aging on explicit memory. PMID- 12735080 TI - Mental work demands protect against cognitive impairment: MAAS prospective cohort study. AB - Little is known about whether persons with mentally demanding jobs are protected against cognitive impairment and whether this association is independent of intellectual abilities and other confounders. Longitudinal data from the Maastricht Aging Study (MAAS) were used to examine this association. After the 1993-1995 baseline examination, there was a first 3-year follow-up examination (1996-1998) among 630 men and women, aged 50 to 80, who exhibited no cognitive impairment at baseline. Persons with mentally demanding jobs had lower risks of developing cognitive impairment during follow-up (36 cases), compared with persons without such jobs (odds ratio = 0.79; 95% confidence interval: 0.65 0.96). About 1.5% of the persons with high mental work demands developed impairment compared to 4% of the persons with few work demands. The protective effect was independent of intellectual abilities and other confounders. Our findings provide evidence that continued and potentially modifiable mental stimulation during adult life may protect men and women against cognitive impairment. PMID- 12735083 TI - Age differences and divided attention: is there a general deficit? AB - It was the goal of this study to determine whether there were age differences specifically associated with the ability to simultaneously execute two tasks, and whether cognitive costs correlated across different situations. Eighty-one young and 86 older adults underwent nine tasks, administered both in single and in dual conditions. Results showed large age differences in raw performances in all conditions. However, a larger cognitive cost in the older adults sample, as assessed by an Age x Condition interaction, was observed only for four out of the nine tasks. Furthermore, age effects were greatly diminished once performance in the single tasks was controlled for. Correlations between the dual tasks, or between the cognitive cost scores, were very low once age was partialled out. Results do not support the notion of general coordination costs and speak against a generalized increase in divided attention costs with advancing age. PMID- 12735081 TI - Percentage decline in masters superathlete track and field performance with aging. AB - Masters athletic records in track and field events, published in September 1999, were analyzed to evaluate the percentage decline in maximum physiological performance with increasing age. Records were normalized using the 30s age records as the baseline and studied through to the 90s age range. Track running records declined with age in a curvilinear fashion [y = 1-exp((T-T0)/tau)] whereas the walking and field events declined in a linear manner [y = alpha (T T'0)]. There were significant differences in the rates of percentage decline in the running events over various distances for both males and females, and significant differences between males and females. Decline with aging was greater for females, and for the longer or endurance running events. There were no differences in the rates of declining function for any of the walking events, and the only jumping event to show a significant difference was the high jump performance, which showed the slowest decline. The walking events declined more slowly than the running events, which declined more slowly than the jumping events. Because of the changes in the weights, heights, and distances at different ages, for both males and females, it was not possible to directly compare rates of decline in the various throwing and hurdling events. The strength-dependent throwing events and the pole vault showed the greatest rates of decline with age. In general terms, men's performance declined to 75% of peak performance in sprint events by the early 70s, in the longer track distances by mid to late 60s, and in field events by mid to late 50s. Women's performance declined to a similar extent by their mid 60s in track events, and by late 40s to early 50s in field events. Some of these differences may be due to differences in training effort and/or other competitive aspects, such as the numbers of athletes involved in the sport, and such differences may be reduced in the future with a more professional approach to these events. PMID- 12735082 TI - Person recognition and approach behaviors among individuals with probable Alzheimer's disease. AB - Sixteen residents at a group home for individuals with probable Alzheimer's disease took part in a person-recognition study. The stimulus was an unfamiliar individual to whom they were exposed for 8 weeks. For nine participants, approach tendency increased from Week 1 to the end of Week 4. Neither total time of exposure to a new person, nor cognitive status (by Mini-Mental State Examination [MMSE] score) predicted changes in approach behaviors over time. An innovative strategy for indexing person recognition is presented for possible use in future research studies with larger samples. PMID- 12735084 TI - Pathological determinants of dementia in Alzheimer's disease (AD). PMID- 12735085 TI - Duplex Doppler ultrasound assessment of clitoral hemodynamics after topical administration of alprostadil in women with arousal and orgasmic disorders. AB - There are limited hemodynamic data in women with arousal or orgasmic disorders and even fewer normative control hemodynamic data in women without sexual dysfunction. In addition, there is limited experience with topical vasoactive agents (used to maximize genital smooth muscle relaxation) applied to the external genitalia during hemodynamic evaluations. The aim of this study was to report duplex Doppler ultrasound clitoral cavernosal arterial changes before and after topical PGE-1 (Alprostadil) administration in control women and in patients with arousal and orgasmic sexual disorders. We found that women with sexual arousal and orgasmic disorders had significantly (p < 0.05) diminished clitoral peak systolic and end diastolic velocity responses compared to controls. Further research is needed to establish the diagnostic role of topical vasoactive agents in the hemodynamic evaluation of women with sexual dysfunction. PMID- 12735086 TI - Genital self-image as a component of sexual health: relationship between genital self-image, female sexual function, and quality of life measures. AB - Thirty-one patients completed a Genital Self-Image Scale along with the Female Sexual Function Index (FSFI; Rosen et al., 2000), Sexual Distress Scale (Derogatis et al., 2001), Perceived Stress Scale (Cohen et al., 1983), Golombok Rust Inventory of Marital State (Rust et al., 1986) Questionnaire, and Beck's Depression Inventory (Beck et al., 1988). A positive genital self-image correlated negatively with amount of sexual distress and depression but not with marital state, perceived stress, or overall sexual function. A positive genital self-image was associated with higher desire but not with arousal, lubrication, orgasm, satisfaction, or absence of pain on the FSFI. Although not correlated directly with sexual function, genital self-image appears to be associated with parameters of the overall sexual experience. PMID- 12735087 TI - Randomized, placebo-controlled, double blind, crossover design trial of the efficacy and safety of Zestra for Women in women with and without female sexual arousal disorder. AB - Zestra for Women is a botanical feminine massage oil formulated to enhance female sexual pleasure and arousal when applied to the vulva. We conducted this randomized, double-blinded, crossover study to evaluate the efficacy and safety of Zestra for Women compared to placebo oil in 10 women with and 10 women without female sexual arousal disorder (FSAD) in conditions of home use in conjunction with sexual activities. Subjects were screened by physical examination, sex therapist interviews, and questionnaires. We randomized qualified subjects to treatment paths and gave them 5 doses of test article and diaries to use at home. At Visit 2, we assessed them by questionnaires and gave them 5 doses of crossover test article and diaries to use at home. At the final visit, we assessed them with questionnaires. We assessed safety by adverse event reports and primary efficacy by responses to a diary question regarding satisfaction with arousal. Secondary efficacy instruments included remaining diary questions, recall-based questionnaires, global assessment questions, and a consumer-testing questionnaire. All 20 subjects completed the study. Three subjects reported single incidences of mild genital burning sensations lasting 5-30 min after use of Zestra for Women. Both normal and FSAD women showed statistically significant improvements, relative to placebo, in level of arousal, level of desire, satisfaction with arousal, genital sensation, ability to have orgasms, and sexual pleasure. Although FSAD women showed greater magnitude of response, the presence of FSAD had no effect on response rates. Zestra for Women was just as effective in women using selective serotonin reuptake inhibitor antidepressants as in women not using antidepressants. Zestra for Women improved sexual function in normal and FSAD women under conditions of home use. PMID- 12735089 TI - Do women gain anything from coitus apart from pregnancy? Changes in the human female genital tract activated by coitus. AB - This article reviews critically studies that describe changes occurring in the female genital tract and pelvic musculature from sexual reflexes activated by stimuli that mimic coitus or induced by sexual arousal. I conclude that apart from the normal erotic pleasure, women gain and maintain vaginal and pelvic functionality from these and, subsequently, from coitus. PMID- 12735088 TI - Characteristics of women with vulvar pain disorders: responses to a Web-based survey. AB - This article presents data contributed by 428 highly educated, internet-savvy women who frequented various vulvar pain discussion lists. The age range was in the reproductive years and older and over 90% were Caucasians. No country of origin was given. They had a number of distressing symptoms, including vulvar pain at rest and with contact, burning, itching, redness, and inflammation. Most felt that they had either vulvar vestibulitis, vulvodynia, or both, although they had other vulvar conditions as well. Many felt that yeast infections, stress, antibiotics, infections, and chemicals played a contributing role. There were a number of comorbidities, including irritable bowel syndrome, fibromyalgia, and interstitial cystitis. Sexual abuse was not a major issue. The vulvar pain destroyed or altered then sex lives, lowered their self-esteem, and affected their relationships. Often, they relied upon understanding partners, support groups, and hobbies but not the medical profession for comfort. PMID- 12735090 TI - Dynamic MR imaging of the sexual arousal response in women. AB - The purpose of our studies was to evaluate whether MR imaging could be used to noninvasively observe and measure the sexual arousal response in normal women. We tested the feasibility as well as the reproducibility of rapid, dynamic, serial high-resolution MR imaging of the genital structures during presentation of neutral and sexually stimulating video material. Results show that these MRI techniques can visualize significant changes in clitoral volume during the stimulus segment of the video presentation. Quantitative measurements made of these changes were robust and reproducible. These studies suggest that MRI techniques may be a useful tool to improve our understanding of the physiology involved with the sexual arousal response in women. These MRI techniques may also prove useful as a surrogate end point marker for testing efficacy of future new treatments for women with sexual arousal disorder. PMID- 12735091 TI - Effects of ovariectomy and estrogen replacement on basal and pelvic nerve stimulated vaginal lubrication in an animal model. AB - The goal of this study was to investigate the effects of ovariectomy and estrogen replacement on vaginal tissue integrity and vaginal lubrication in basal conditions and in response to pelvic nerve stimulation (PNS). Two weeks after ovariectomy, female New Zealand White rabbits were administered vehicle or estradiol (200 micrograms/day) for an additional 2 weeks. Ovariectomy caused significant vaginal atrophy and diminished vaginal lubrication in the basal state and after PNS, compared to intact controls. Estrogen replacement normalized lubrication values and tissue wet weight to control levels. In conclusion, vaginal tissue integrity and lubrication are diminished by ovariectomy and are normalized by estrogen replacement. PMID- 12735092 TI - A prospective duplex Doppler ultrasonographic study in women with sexual arousal disorder to objectively assess genital engorgement induced by EROS therapy. AB - The EROS therapy device is FDA-approved for the treatment of women with sexual dysfunction and has been shown to improve sexual function and satisfaction. The aim of this study was to obtain objective information regarding the ability of the EROS Therapy device to induce clitoral and corpus spongiosum volumetric and hemodynamic changes following therapeutic use in women with sexual arousal disorder. Seven patients with sexual arousal disorder formed the study population. All seven subjects met inclusion and exclusion criteria, including having normal hormonal values at the time of the study. All seven subjects were able to comfortably operate the device. All seven reported either slight-to moderate pleasure or orgasm at home with the device. We observed no adverse events. This study shows that EROS therapy is associated with significant increases in clitoral and corpus spongiosum diameter as well as with clitoral and corpus spongiosum peak systolic and end-diastolic velocity values. PMID- 12735093 TI - Serum allopregnanolone levels relate to FSFI score during the menstrual cycle. AB - The aim of this study was to measure serum allopregnanolone levels and other hormones in women who completed the Female Sexual Function Index (FSFI; Rosen et al., 2000) during the follicular or the luteal phase of the menstrual cycle. Twenty-nine women with a regular menstrual cycle completed the FSFI during days 5 7 and days 19-21 of their menstrual cycles. We collected a blood sample on the same days so that we could determine levels of allopregnanolone (AP), dehydroepiandrosterone (DHEA), free testosterone (FreeT), androstenedione (A), and estradiol (E2); we stored serum at--20 degrees C until we assayed it. We performed statistical analyses by parametric and nonparametric comparisons and correlations, as appropriate. We found that the full FSFI score was 23 +/- 11.3 (mean +/- SD; 31% under 95% confidence interval = 18.7) in our study population. We also found a significant positive correlation between serum allopregnanolone levels and each FSFI domain, including full scale score (r = 0.47, p < 0.01), with the exception of pain. Similarly, we found a positive correlation between FreeT and each FSFI domain and full scale score (r = 0.55, p < 0.002). We found no significant correlation among FSFI scores and DHEA, A, or E2 plasma levels. It is interesting to note that FSFI full score was significantly higher (p < 0.04) in women tested in the luteal phase compared to women tested in the follicular phase. Serum allopregnanolone may be relevant to female sexuality directly or by its influence on a woman's general sense of well-being during the menstrual cycle. Further studies are required to test the hypothesis that high serum allopregnanolone levels in the luteal phase play a role in women's attitudes toward self-report questionnaires on sexual function. PMID- 12735094 TI - Electrical stimulation (ES) in the management of sexual pain disorders. AB - We performed an open study to investigate the use of electrical stimulation (ES) on the vestibular area and vaginal introitus in women with sexual pain disorders. We recruited 29 women (age range 20-45 years) from among the patients at our Reproductive Psychobiology Unit to participate in the present study. They each experienced vestibular pain, inducing dyspareunia and vaginism. We performed ES with an ECL43400 apparatus (Elite, EssediEsse srl, Milan, Italy) once a week for 10 weeks. To evaluate the muscular activity of the perineal floor and sexual function, we employed the same apparatus with a vaginal probe for recording myoelectrical activity (muV), we employed a VAS scale for evaluating pain, and we administered the Female Sexual Function Index (FSFI; Rosen et al., 2000) before and after the study protocol. We analyzed data by parametric and nonparametric comparisons and correlations, as appropriate. Our major findings were as follows: (a) the contractile ability of pelvic floor muscles (p < 0.001), as well as the resting ability (p < 0.001), significantly improved following ES; (b) the current intensity tolerated significantly increased (p < 0.001) throughout the study, from 41.3 +/- 7.4 mA at the start of the study to 50 +/- 7.4 mA at the end of the stimulation protocol; (c) the Visual Analogic Scale (VAS) for pain significantly declined (p < 0.001), whereas FSFI pain scores (p < 0.001) and full scale scores (p < 0.001) significantly improved following ES, and 4 out of 9 women with vaginism went back to coital activity; (d) FSFI pain score and the current intensity tolerated, both before (R = .59; p < 0.006) and at the end (R = .53; p < 0.02) of the stimulation protocol, positively correlated. ES may be effective in the management of sexual pain disorders. Further controlled studies are necessary to standardize stimulation protocols according to the severity of pain and to better clarify the long-term clinical effects of ES. PMID- 12735095 TI - Pain measurement in vulvodynia. AB - Current approaches to the conceptualization of vulvodynia focus either on issues of sexuality or underlying pathophysiology but tend to neglect the central symptom of pain. An adequate understanding of this condition will not be achieved, however, without examining all three factors and how they interact. A multiaxial pain approach has provided data regarding the intensity, location, temporal pattern, underlying system, and sensory quality of vulvodynic pain. In addition, standardized vulvar pain measurement techniques, including a new device called a vulvalgesiometer, have been developed, making possible the collection of reliable pain ratings. To date, this approach has been promising for the differential diagnosis, classification, and understanding of vulvar pain conditions. PMID- 12735097 TI - EHP Toxicogenomics: a publication forum in the postgenome era. PMID- 12735098 TI - The new biology. PMID- 12735096 TI - Sexuality, intimacy, and gynecological cancer. AB - On a psychological level, not all changes in sexual functioning following gynecological cancer treatment automatically lead to sexual problems or dysfunctions. Whether sexual dissatisfaction occurs will also depend on personal factors, social factors, and the context in which these negative changes occur. PMID- 12735099 TI - HapMap: building a database with blocks. PMID- 12735100 TI - National Center for Toxicogenomics: an introduction. PMID- 12735101 TI - Toxicogenomics. An emerging discipline. PMID- 12735102 TI - Liver library. Creating a microarray for hepatotoxicants. PMID- 12735103 TI - Monitoring estrogenic effects. A genomics approach. PMID- 12735104 TI - The human proteome organization (HUPO) and environmental health. AB - The Human Proteome Organization, or HUPO, was formed to promote research and large-scale analysis of the human proteome. By consolidating national proteome organizations into an international body, HUPO will coordinate international initiatives, biological resources, protocols, standards and data for studying the human proteome. HUPO has identified five key areas to advance study of the human proteome, specifically in bioinformatics, new technologies, the plasma proteome, cell models, and a public antibody initiative. Consideration of three major issue areas may help develop HUPO's strategy for human proteome study. First is the need to distinguish the value of high throughput platforms from discovery platforms in proteomics. Second is the importance for international planning on integrating both transcriptome and proteome data and databases. Last is that effects of the environment from chemical, physical, and biological exposures alter the expression and structure of the proteome, which become manifest in long term adverse health effects and disease. Environmental health research stands to greatly benefit from the shared resources, data, and vision of the HUPO organization as a valuable resource in exploiting knowledge of the human proteome toward improving public health. PMID- 12735105 TI - Application of toxicogenomic analysis to risk assessment of delayed long-term effects of multiple chemicals, including endocrine disruptors in human fetuses. AB - Our previous studies analyzing umbilical cords show that human fetuses in Japan are exposed to multiple chemicals. Because of these findings, we believe it is necessary to establish a new strategy for examining the possible delayed long term effects caused by prenatal exposure to multiple chemical combinations and evaluating the health risk to human fetuses. In this commentary we describe our attempts to apply toxicogenomic analysis of umbilical cords, using DNA microarray for future risk assessment. Because the umbilical cord is part of the fetal tissue, it is possible to estimate the effects of chemicals on the fetus by analyzing alteration of the gene expression. This type of toxicogenomic analysis could be a powerful and effective tool for developing a new risk assessment strategy to help investigators understand and possibly prevent long-term effects caused by fetal exposure to multiple chemicals. Worldwide cooperation is needed to establish a new stragegy for risk assessment using toxicogenomic analysis that focuses on the human fetus. PMID- 12735106 TI - Systems toxicology and the Chemical Effects in Biological Systems (CEBS) knowledge base. AB - The National Center for Toxicogenomics is developing the first public toxicogenomics knowledge base that combines molecular expression data sets from transcriptomics, proteomics, metabonomics, and conventional toxicology with metabolic, toxicologcal pathway, and gene regulatory network information relevant to environmental toxicology and human disease. It is called the Chemical Effects in Biological Systems (CEBS) knowledge base and is designed to meet the information needs of "systems toxicology," involving the study of perturbation by chemicals and stressors, monitoring changes in molecular expression and conventional toxicological parameters, and iteratively integrating biological response data to describe the functioning organism. Based upon functional genomics approaches used successfully in analyzing yeast gene expression data sets, relational and descriptive compendia will be assembled for toxicologically important genes, groups of genes, single nucleotide polymorphisms (SNPs), and mutant and knockout phenotypes. CEBS data sets will be fully documented in the experimental protocol and therefore searchable by compound, structure, toxicity end point, pathology and point, gene, gene group, SNP, pathway, and network as a function of dose, time, and the phenotype of the target tissue. A knowledge base is being developed by assimilating toxicological, biological, and chemical information from multiple public domain databases and by progressively refining that information about gene, protein, and metabolite expression for classes of chemicals and their biological effects in various species. By analogy to the GenBank database for genome sequences, researchers will globally query (or BLAST) CEBS using a transcriptome of a tissue of interest (or a list of outliers) to have the knowledge base return information on genes, groups of genes, metabolic and toxicological pathways, and contextually associated phenotypic information for compounds that display similar response profiles. With high-quality data content, CEBS will ultimately become a resource to support hypothesis-driven and discovery research that contributes effectively to drug safety and the improvement of risk assessments for chemicals in the environment. The CEBS development effort will span a decade or more. PMID- 12735107 TI - Expression profiling of estrogenic compounds using a sheepshead minnow cDNA macroarray. AB - A variety of anthropogenic compounds are capable of binding to the estrogen receptor (ER) of vertebrate species. Binding of these chemicals to the ER can interfere with homeostasis by altering normal gene expression patterns. The purpose of this study was to characterize the expression of 30 genes using a sheepshead minnow (Cyprinodon variegatus) cDNA macroarray. Many of the genes on the array were previously identified by differential display reverse transcriptase-polymerase chain reaction to be upregulated or downregulated in sheepshead minnows treated through aqueous exposure to known or suspected estrogenic chemicals. The results of this study show that 17 beta-estradiol (E2), 17 alpha-ethinyl estradiol (EE2), diethylstilbestrol (DES), and methoxychlor (MXC) have similar genetic signatures for the 30 genes examined. The genetic signature of fish treated with p-nonylphenol was identical in pattern to that in fish treated with E2, EE2, DES, and MXC except for the additional upregulation of a cDNA clone that shares similarity to ubiquitin-conjugating enzyme 9. Endosulfan produced results that resembled the gene expression patterns of untreated control fish with exception of the upregulation of estrogen receptor alpha and the downregulation of a cDNA clone that shares similarity to 3-hydroxy-3 methylglutaryl-coenzyme A reductase. We show that our estrogen-responsive cDNA macroarray can detect dose-dependent changes in gene expression patterns in fish treated with EE2. PMID- 12735108 TI - Gene expression profiles associated with inflammation, fibrosis, and cholestasis in mouse liver after griseofulvin. AB - Erythropoietic protoporphyria patients can develop cholestasis, severe hepatic damage, fibrosis, and cirrhosis. We modeled this hepatic pathology in C57BL/6J and BALB/c mice using griseofulvin and analyzed 3,127 genes for alteration of expression in the liver before and during the onset of protoporphyria, cholestasis, inflammation, and hepatic fibrosis. The two mouse strains developed different levels of pathologic damage in response to the griseofulvin. Characteristic gene expression profiles could be associated with griseofulvin induced gene expression, disruption of lipid metabolism, and the pathologic states of inflammation, early fibrosis, and cholestasis. Additionally, some genes individually indicated an alteration of homeostasis. or pathologic state; for example, fibroblast proliferation was potentially indicated by increased calcyclin (SA100a6) expression. Changes in cytochrome P450 (Cyp) gene expression were particularly pronounced, with increased expression of the Cyp2a, Cyp2b, and Cyp3a families. Decreased Cyp4a10 and Cyp4a14 expression was observed that could be associated with early pathologic change. A potential decrease in bile acid and steroid biosynthesis was indicated by the decreased expression of Cyp7b1 and Hsd3b4, respectively. DNA damage was indicated by induction of GADD45. This study illustrates how transcriptional programs can be associated with different stimuli in the same experiment. The time course of change in the gene expression profile compared with changes in pathology and clinical chemistry shows the potential of this approach for modeling causative, predictive, and adaptive changes in gene expression during pathologic change. PMID- 12735109 TI - Intrinsic hepatic phenotype associated with the Cyp1a2 gene as shown by cDNA expression microarray analysis of the knockout mouse. AB - Several forms of cytochrome P450 (CYP) appear to metabolize principally pharmaceutical agents, as well as other dietary and plant chemicals. Other CYP forms have major roles in steroid, sterol, and bile acid metabolism. CYP1A2 expression is constitutively high in mouse liver and is well known for metabolizing several drugs and many procarcinogens to reactive intermediates that can cause toxicity or cancer. CYP1A2 is also known to carry out several endogenous functions such as uroporphyrinogen and melatonin oxidation and the 2- and 4-hydroxylations of estradiol. We have used cDNA microarray analysis of the untreated Cyp1a2(-/-) knockout mouse to search for changes in gene expression that might indicate important intrinsic roles for this enzyme. For 15 of the up- or downregulated genes, these increases or decreases were corroborated by reverse transcription real-time polymerase chain reaction. Other than upregulation of the Hprt gene (used in the selection procedure for disrupting the Cyp1a2 gene), we found several genes upregulated that are associated with cell-cycle regulation and lipid metabolism. Besides Cyp1a2, the gene exhibiting the greatest downregulation was Igfbp1 (insulin-like growth factor binding protein-1), showing only 12% expression of that in the Cyp1a2(+/+) wild-type liver. Recurrent themes between both up- and downregulated genes include cell-cycle control, insulin action, lipogenesis, and fatty acid and cholesterol biosynthetic pathways. Histologically, the Cyp1a2(-/-) mouse exhibited an approximately 50% decrease in lipid stored in hepatocytes, and 50% increase in lipid present in interstitial fat-storing cells compared with that in the Cyp1a2(+/+) wild-type. These data suggest that the CYP1A2 enzyme might perform additional hepatic endogenous functions heretofore not appreciated. PMID- 12735110 TI - Development of a DNA microarray for toxicology based on hepatotoxin-regulated sequences. AB - Toxicogenomics is an emerging field combining genomics and bioinformatics to identify and characterize mechanisms of toxicity of compounds. One of the main tools used in toxicogenomics is DNA microarrays. We have used a novel strategy to create a library highly enriched for genes expressed in the liver under hepatotoxic conditions. Using this library, we have created a new oligonucleotide microarray dedicated to the study of rat liver function. Oligonucleotide probes for these genes were designed and used in experimental hybridization studies to deduce the correct sequence orientation and to determine those sequences exhibiting differential regulation under a variety of toxicity-related treatments and conditions. The final array was benchmarked on treatments with 3 methylcholanthrene, Aroclor 1254, and cyclopropane carboxylic acid. Our results showed that the subtractive hybridization greatly enriched for genes regulated during a hepatotoxic response. Overall, our strategy for design of a new rat toxicology microarray can be applied to other systems as well and should aid greatly in the development of microarrays targeted for specific scientific areas. PMID- 12735111 TI - Exploiting genome data to understand the function, regulation, and evolutionary origins of toxicologically relevant genes. AB - The wealth of new information coming from the many genome sequencing projects is providing unprecedented opportunities for major advances in all areas of biology, including the environmental health sciences. To facilitate this discovery process, experts in the fields of functional genomics and informatics and the emerging field of toxicogenomics recently gathered at the Mount Desert Island Biological Laboratory in Salisbury Cove, Maine, site of a National Institute of Environmental Health Sciences Marine and Freshwater Biomedical Science Center, to share their ideas and latest research findings. The goal of the symposium was to highlight approaches that may be used to identify and characterize toxicologically relevant genes being discovered in the genome sequencing projects. Many of the approaches rely heavily on comparative models as a way of identifying gene homology, ontology, and physiologic function, and on the availability of databases that facilitate storage, analysis, interpretation, and widespread dissemination of relevant data. PMID- 12735112 TI - Normative aging of the respiratory system. AB - An absolute quantified normal rate of change and normal range of functions of the respiratory system applicable to all older adults as they age is elusive. Like life expectancy, which is dependent on a cohort effect, the norms of respiratory system function are related to the birth cohort to which a given individual belongs and the age at which the parameter is assessed. No single rate of change can express normal across all age ranges even for those individuals in apparently good health [29]. Analogous to defining risk factors for a disease, determining that a change in anatomy or physiology is not disease requires stringent prospective evaluation for the absence of occult disease and known risk factors for disease prior to concluding that the alteration is inevitable with the normal aging process [19,31]. Additional limitations in quantifying the norms of respiratory function with age are the lack of participation of the oldest adults in studies and the lack of precision and accuracy in these performance-based measurements. The data, although limited, do support a qualitative emphysematous change in lung histology and lung-thorax mechanics. This change plus altered lung volumes influence oxygenation and oxygen consumption. There is no evidence that the changes in the respiratory system with aging impact day-to-day function of older adults, but they may become evident under circumstances when physiologic demand reaches the limits of supply. Despite changes in cholinergic and adrenergic receptor functioning, there is no evidence to suggest altering prescribing these classes of medications for older people. Pioneer physiologists asked the original question "Is there a difference in this measurement for older people?" Researchers in pulmonary medicine, pathology, radiology, epidemiology, and public health have continued to revise the question toward the clinical implications while studying the aging process from their respective viewpoints. Clinicians who need to develop an integrated care plan should neither rely on formulas to "normalize" a measurement for age nor assume that a established predictive value of a diagnostic test done in young adults can be automatically applied to geriatric patients [4]. Rather, the clinical situation should consider that the variability in normal is greater with older age and that all diagnostic tests and care plans should be considered in the context of the patient's symptoms [5]. PMID- 12735114 TI - Preoperative pulmonary assessment of the older adult. AB - Postoperative pulmonary complications in the elderly are common and are a significant source of morbidity, mortality, and prolonged length of stay. Risk factors differ from the well-known risk factors for cardiac complications and can be divided into patient- and procedure-related factors. Patient-related factors include COPD, recent cigarette use, poor general health status as defined by Goldman or ASA class, dependent functional status, and laboratory parameters including abnormal chest radiograph, renal insufficiency, and low serum albumin. Age is a minor risk factor when adjusted for comorbidities and confers approximately a two-fold increase in risk. Elderly patients who are otherwise acceptable surgical candidates should not be denied surgery based solely on age and concern for postoperative pulmonary complications. The surgical site is the single most important predictor of pulmonary complications. High-risk surgeries include thoracic, upper abdominal, aortic, neurosurgery, and peripheral vascular. Other procedure-related risk factors include surgery lasting longer than 3 hours, the use of general anesthesia, pancuronium use, and emergency surgery. Clinicians should not recommend routine preoperative spirometry before high-risk surgery because it is no more accurate in predicting risk than clinical evaluation. Patients who might benefit from preoperative spirometry include those who have unexplained dyspnea or exercise intolerance and those who have COPD or asthma in whom uncertainty exists as to the status of airflow obstruction when compared with baseline. After identifying patients at risk for postoperative pulmonary complications, clinicians can recommend strategies to reduce risk throughout the operative period. In addition to minimizing or avoiding the above risk factors, optimization of COPD or asthma, deep breathing exercises, incentive spirometry, and epidural local anesthetics reduce the risk of postoperative pulmonary complications in elderly surgical patients. PMID- 12735113 TI - Evaluation of dyspnea in the elderly. AB - The word "dyspnea" encompasses many different features. It can be considered to be a sensation, a symptom, or an illness. From a practical perspective many physicians and nurses use dyspnea to refer to difficult or labored breathing or an uncomfortable awareness of breathing. The American Thoracic Society recently defined dyspnea as a subjective experience of breathing discomfort that consists of qualitatively distinct sensations that vary in intensity. PMID- 12735115 TI - Asthma in the elderly. AB - Asthma is common in the elderly population and the differences between younger and older asthmatics should be appreciated (Table 2). Asthma is frequently overlooked in the geriatric population. Objective measures of pulmonary function can aid in a prompt diagnosis and lead to effective treatment and improved quality of life. Because smoking is an important risk factor for asthma-like symptoms of wheezing, cough, and sputum production, asthma is frequently confused with COPD. When airflow obstruction is found, attempts to demonstrate reversibility can uncover an asthmatic component to the disease. In patients who have asthma symptoms and no airflow obstruction, methacholine testing is helpful. When a normal methacholine challenge is present, a diagnosis of asthma can be excluded and the physician can pursue other diagnostic considerations such as heart failure, chronic aspiration syndrome, pulmonary embolic disease, and carcinoma of the lung. The onset of wheezing, shortness of breath, and cough in an elderly patient is likely to cause concern. Although the adage "all that wheezes is not asthma" is true at any age, it is especially true in the elderly. Diagnosis based on objective measures is essential. PMID- 12735116 TI - Smoking cessation in the elderly. AB - Today, two generations after the first Surgeon General's report, and with abundant evidence of the catastrophic consequences of smoking, no serious person can be unaware that smokers risk their lives and health. We also know that quitting smoking--at any age--promises significant health benefits. When offered the tools they need, older smokers quit smoking at rates comparable to those of younger smokers despite their skepticism, fatalism, and self-doubt. Older smokers should be encouraged to enter programs that stress the health benefits derived and identify the risks they are avoiding by quitting smoking. These programs establish quit dates, use sound behavioral modification techniques, provide strategies for stress management and relaxation, treat withdrawal symptoms, and provide regular and continuing follow-up. The patient is asked to make an existential change, and the physician should provide encouragement and promote self-confidence by emphasizing that, despite setbacks, with repeated efforts, success can be achieved. Clinicians can influence patients to quit smoking, and they should. PMID- 12735117 TI - Community-acquired pneumonia in elderly patients. AB - CAP in elderly patients carries a significant economic and clinical burden and will be more commonly encountered in the future as the US population ages. Diagnosis may be obscured by a nonclassic presentation in an elderly patient, and the clinician needs to be especially suspicious of pneumonia whenever the clinical status of an elderly patient deteriorates. The single most important clinical decision is the site of care; this determination is not always based on clinical factors but also on social factors. Severity assessment is key to stratifying appropriate therapy and to predicting outcome. Timely and appropriate empiric therapy enhances the likelihood of a good clinical outcome, although clinical resolution may be more delayed than in younger patients. Newly emerging patterns of antibiotic resistance have altered recent guidelines for CAP treatment; DRSP is now a consideration in elderly patients because an age older than 65 years is a well-described risk factor for infection with this organism. Prevention should always be implemented, with a focus on pneumococcal and influenza vaccination. PMID- 12735118 TI - Tuberculosis in the elderly. AB - As the result of the strengthening of TB-control programs nationwide, a decline in the overall number of reported TB cases in the United States has been observed within the last 10 years. Despite these declines in absolute numbers, the elderly continue to account for a disproportionate share of the cases. The high number of cases diagnosed at autopsy among the elderly suggests that this condition often remains unrecognized, possibly due to the subtle clinical manifestations in this age group. Evidence suggests that, compared with their community-dwelling counterparts, the institutionalized elderly are at a greater risk for re activation of latent TB and for the acquisition of new TB infection. More studies are needed to make final conclusions. New guidelines for the treatment of LTBI emphasize targeted TST among persons at high risk for development of active TB and no longer use age as an exclusionary condition. All nursing home residents must therefore be regularly screened for LTBI and treated if necessary. Even though elderly persons are at greater risk for hepatic toxicity from TB treatment, the poor outcome of untreated TB in this age group warrants more aggressive treatment of this condition. PMID- 12735119 TI - Lung cancer in elderly patients. AB - Lung cancer is one of the most common causes of death in elderly patients in the United States. Treatment advances have improved survival in selected patients. The available treatments carry the risk of morbidity and mortality but the benefit in most patients far outweighs the risks, given the dismal prognosis of untreated disease. Elderly patients with lung cancer need careful attention during pretreatment assessment. Advanced age alone, however, should not contraindicate aggressive treatment. In the high-risk groups it is important to involve a team of physicians including surgeons, radiation oncologists, medical oncologists, and pulmonologists, who are familiar with current treatment options and their risks. PMID- 12735120 TI - Diagnosis and treatment of pulmonary embolism in the elderly. AB - Pulmonary embolism (PE) is a difficult diagnosis in patients of all ages, but more so in the elderly. Nonspecific symptoms and laboratory results are often misattributed to common diseases or to age itself, and can delay or even deter the diagnosis and treatment of PE. Advanced age is sometimes mistakenly seen as a contraindication to anticoagulation and thrombolysis. Together, these factors contribute to the higher morbidity and mortality associated with PE in the elderly than in younger patients. This article reviews the risk factors, diagnosis, and treatment of PE as it applies to the elderly. PMID- 12735121 TI - Sleep in the elderly. What is normal? AB - Sleep is a basic biologic function that changes with normal aging and in many pathologic states. Some of the changes with aging are so profound that it is difficult to separate normal aging from disease. The problem is made worse by the difficulty of recognizing many common sleep disorders. Complaints of poor sleep or daytime somnolence are common in all adults but are more prevalent in elderly individuals. This article addresses normal sleep and the changes expected with aging, and reviews the more common sleep diseases in the elderly population, such as insomnia, sleep-disordered breathing, periodic limb movements of sleep, and the rapid eye movement sleep-behavior disorder. PMID- 12735122 TI - Pulmonary physical medicine interventions for elderly patients with muscular dysfunction. AB - The respiratory consequences of respiratory muscle impairment, with or without bulbar muscle weakness, usually involve inadequate ventilation and airway secretion encumbrance and may progress to respiratory failure. This article focuses on muscular dysfunction of the bulbar and respiratory musculature in elderly patients with neuromuscular diseases or central nervous system disorders. In addition, the article discusses application of pulmonary physical medicine principles for patients with predominantly restrictive pulmonary disorders caused primarily by muscular dysfunction. Diagnostic evaluation, respective clinical courses and responses to therapy, and the use of noninvasive ventilatory support outside of an acute care setting are also reviewed. PMID- 12735123 TI - Respiratory failure in elderly patients. AB - Elderly individuals comprise an increasing proportion of the population and represent a progressively expanding number of patients admitted to the ICU. Because of underlying pulmonary disease, loss of muscle mass, and other comorbid conditions, older persons are at increased risk of developing respiratory failure. Recognition of this vulnerability and the adoption of proactive measures to prevent decompensation requiring intrusive support are major priorities together with clear delineation of patients' wishes regarding the extent of support desired should clinical deterioration occur. Further, the development of coordinated approaches to identify patients at risk for respiratory failure and strategies to prevent the need for intubation, such as the use of NIV in appropriate patients, are crucial. As soon as endotracheal intubation and mechanical ventilation are implemented strategies that facilitate the liberation of elderly patients from the ventilator are especially important. The emphasis on a team approach, which characterizes geriatric medicine, is essential in coordinating the skills of multiple health care professionals in this setting. Respiratory failure can neither be effectively diagnosed nor managed in isolation. Integration with all other aspects of care is essential. Patient vulnerability to nosocomial complications and the "cascade effect" of these problems such as the effects of medications and invasive supportive procedures all impact on respiratory care of elderly patients. For example, prolonged mechanical ventilation may be required long after resolution of the underlying cause of respiratory failure because of unrecognized and untreated delirium or residual effects of small doses of sedative and/or analgesic agents or other medications in elderly patients with altered drug metabolism. The deleterious impact of the foreign and sometimes threatening ICU environment and/or sleep deprivation on the patient's course are too often overlooked because the physician focuses management on physiologic measurements, mechanical ventilator settings, and other technologic nuances of care [40]. Review of the literature suggests that the development of respiratory failure in patients with certain disease processes such as COPD, IPF, and ARDS in elderly patients may lead to worsened outcome but it appears that the disease process itself, rather than the age of the patient, is the major determinant of outcome. Additional studies suggest that other comorbid factors may be more important than age. Only when comorbid processes are taken into account should decisions be made about the efficacy of instituting mechanical ventilation. In addition, because outcome prediction appears to be more accurate for groups of patients rather than for individual patients a well-structured therapeutic trial of instituting mechanical ventilation, even if comorbidities are present, may be indicated in certain patients if appropriately informed patients wish to pursue this course. This approach requires careful and realistic definition of potential outcomes, focus on optimizing treatment of the reversible components of the illness, and continuous communication with the patient and family. Although many clinicians share a nihilistic view regarding the potential usefulness of mechanical ventilation in elderly patients few data warrant this negative prognostication and more outcome studies are needed to delineate the optimum application of this element of supportive care. As with other interventions individualization of the decision must take into account the patient's premorbid status, concomitant conditions, the nature of the precipitating illness and its prospects for improvement, and most important, patient preferences. In this determination pursuing the course most consistent with the patient's wishes is essential and it must be appreciated that caregivers' impressions regarding the vigor of support desired by the patient are often erroneous. The SUPPORT investigators observed that clinicians often underestimated the degree of intervention desired by older patients assuming that less care would be desired [13]. Thus, as in other circumstances, effective communication and elicitation of patients' preferences regarding management options is crucial in the management of respiratory failure. The frequent discordance between patient preferences and the wishes of family members or other surrogate decision makers impose major clinical challenges and also mandates further investigation. PMID- 12735124 TI - Managing respiratory symptoms at the end of life. AB - Early and clear discussion and articulation of preferences about interventions with increasing burdens and diminishing benefits is helpful in identifying the goals of care and planning management for patients who have unremitting terminal illnesses. The development of respiratory symptoms such as dyspnea, cough, and hiccups is common and can often be anticipated. Aggressive evaluation and treatment should be pursued and offered to palliate symptoms at the end of life. PMID- 12735126 TI - Pathologic spectrum and prognostic significance of underlying liver disease in hepatocellular carcinoma. AB - Although controversy persists about the influence of cirrhosis on the incidence of portal vein invasion and other prognostic indicators of recurrence and survival (e.g., histologic grade, mitotic activity, multiplicity), it is clear that the degree of cirrhosis is important for the long-term survival of patients with HCC. Multicentricity is especially important because it reflects the field carcinogenesis associated with viral hepatitis and particularly chronic HCV infection [59]. A better understanding of the different mechanisms linked to tumor recurrence will help select the best candidates for curative surgery and help tailor adjuvant therapy, such as interferon therapy, to each patient [60]. Finally, the importance of vascular invasion, number of tumors, and tumor size of HCCs in addition to the effect of fibrosis has led Vauthey et al [20] to propose a simplified staging of HCC with better prognostication of survival. PMID- 12735125 TI - What surgeons should know about viral hepatitis and hepatocellular carcinoma. AB - Most patients with hepatocellular carcinoma (HCC) have viral hepatitis, either hepatitis C (HCV) or hepatitis B (HBV). HCV is an RNA virus that does not integrate into the host genome but likely induces HCC through viral protein: for example, host protein interactions or via the inflammatory response to the virus. Eradication of HCV with interferon plus riboviron therapy may help prevent cancer recurrence in selected patients. In contrast to HCV, HBV is an DNA virus that integrates into the host genome, and this integration is believed, in part, to be carcinogenic. HBV is usually classified as replicative (DNA-positive in the serum) or nonreplicative (DNA-negative in the serum). Treatment with nucleoside analogs is indicated in most patients with cirrhotic-stage replicating HBV. Nonreplicative stages of this virus do not merit therapy with these agents. PMID- 12735127 TI - Spectrum and significance of microscopic vascular invasion in hepatocellular carcinoma. AB - Microscopic vascular invasion frequently occurs in patients with hepatocellular carcinoma (HCC). It is a biologic characteristic of HCC to consider when selecting patients for surgical treatment. In addition, microscopic vascular invasion is important to consider in the development of strategies to prevent cancer recurrence after potentially curative therapy. PMID- 12735128 TI - Evaluation of the new AJCC/UICC staging system for hepatocellular carcinoma after hepatic resection in Chinese patients. AB - Hepatic resection is considered the treatment of choice for HCC; however, the prognosis of patients after resection of HCC varies widely, depending on the clinicopathologic features. The American Joint Committee on Cancer (AJCC)/International Union Against Cancer (UICC) tumor-node-metastasis (TNM) staging system is widely used evaluating patients with HCC. The current TNM staging for HCC uses pathologic characteristics of tumors, including size, number and location of tumor nodules, presence of vascular invasion, perforation of visceral peritoneum, and invasion of adjacent organs as criteria for T staging. Recently, a simplified AJCC/UICC staging for HCC has been proposed. In addition, the Liver Cancer Study Group of Japan proposed a new simplified staging system based on number of tumor nodules, size of tumors, and invasion into the portal vein, hepatic vein, or bile duct. This article evaluates the prognostic value of the new AJCC/UICC TNM staging and the new Japanese staging in a large cohort of Chinese patients who underwent hepatectomy for HCC in a single institution. PMID- 12735129 TI - Selective approach to major hepatic resection for hepatocellular carcinoma in chronic liver disease. AB - For large HCCs, partial liver resection remains the best therapeutic option for cure because neither liver transplantation nor percutaneous treatments are indicated. In specialized centers, a better selection of at-risk patients and technical procedures, including the use of intermittent inflow occlusion and the anterior approach, have contributed to improve dramatically the outcome of major liver resection for HCC in CLD. In addition, portal vein embolization has become an important tool to hypertrophy the future liver remnant before major liver resection in cirrhotic patients with apparently normal liver function. PMID- 12735130 TI - Results of hepatic resection for hepatocellular carcinoma invading major portal and/or hepatic veins. AB - Nonsurgical therapy for patients with advanced hepatocellular carcinoma (HCC) has yielded poor long-term survival. This study evaluates the effects of surgical treatments for patients with HCC invading major portal and/or hepatic veins. The surgical results of 112 patients with HCC invading major portal and/or hepatic veins who underwent hepatic resection between 1985 and 2001 were studied to evaluate the feasibility of hepatic resection as a local treatment. PMID- 12735131 TI - Indications and limitations of liver transplantation for hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is a common cause of cancer-related death worldwide, yet remains difficult to treat, with dismal overall long-term survival rates. Recent strategies using liver transplantation for carefully selected patients with stage I and II HCC and cirrhosis have shown promising results, with 5-year survival rates comparable to survival rates for transplantation patients without malignancy. Currently, however, limited resources and a severe organ shortage make liver transplantation an option for only a limited number of patients with HCC in the United States. Future studies must document the long term success of this therapy and improve methods for disease control before and after transplantation. PMID- 12735132 TI - Intraoperative ultrasonography in liver cancer. AB - IOUS has become increasingly important for surgical resection in patients with cirrhosis and healthy liver. IOUS is important in the diagnosis and staging of liver cancer and as an element of the surgical technique, and IOUS can now be considered a fundamental tool for hepatobiliary and other surgical procedures [3]. The American College of Surgeons has recently recognized the need for surgeons to have specific training in ultrasonography. Meanwhile, dedicated monographs on IOUS have been published in the United States, Chile, and Europe [39-42]. PMID- 12735133 TI - Hepatic artery embolization for hepatocellular carcinoma: technique, patient selection, and outcomes. AB - Most patients with HCC do not qualify for surgical interventions. In carefully selected patients, TACE may improve survival, reduce the rate of tumor growth, and decrease the incidence of portal vein occlusion. Since the introduction of TACE in the 1980s, the technical aspects of the procedure have significantly improved. Sophisticated angiographic equipment and techniques have made superselective arterial catheterization possible for more focused drug delivery. The use of ethiodized oil allows for more effective targeting of HCC and provides dual embolization of the hepatic artery and the portal venules supplying the tumor. Many important technical questions about TACE remain unanswered at this time: there are no reliable, standardized patient selection criteria, ideal cytotoxic agents have not yet been identified, the optimal dose of ethiodized oil has not been confirmed, and the optimal frequency and timing of repeat treatment sessions remain unknown. One major limitation of TACE--the need for repeated treatments, which can result in deterioration of liver function--may be avoided by use of a combination of interventional therapies. The combination of limited TACE with PEI or RFA may lead to improved survival and decreased risk of liver failure. More recently, two excellent randomized clinical trials have demonstrated significant survival benefit for patients treated with TACE when compared with those treated symptomatically. PMID- 12735134 TI - Are there indications for chemotherapy in hepatocellular carcinoma? AB - Single-agent Adriamycin gives a response rate of around 15% to 20%, and with combination therapy, this figure rises to around 20% to 35%; however, there is no proven survival benefit. Nonetheless, HCC is clearly not entirely chemotherapy resistant and complete pathologic remission is possible after systemic combination chemotherapy alone. Major methodologic problems remain in assessment of response and the survival benefit from systemic therapy. Until these are resolved, the answer to the question posed in this article is unknown; further appropriate studies remain to be undertaken. PMID- 12735135 TI - Biology of colorectal liver metastases. AB - The elucidation of the interaction of the tumor and its microenvironment has increased surgeons' understanding of the biologic mechanisms mediating metastases formation. The ability to develop effective biologic therapies will depend on understanding the complex interactions among endothelial cells and tumor cells and on the cytokine cross-talk among all cells within a particular tumor at a particular site. Biologic therapies must be targeted to both the site and the tumor. By the time metastases form, most steps in the metastatic cascade have been completed. Therapy to down-regulate or interrupt the last stages of metastases, proliferation, and angiogenesis and mechanisms to disrupt cell survival signals appear to be the most promising areas of investigation. PMID- 12735136 TI - Angiogenesis and hepatic colorectal metastases. AB - Angiogenesis is a critical step in the metastatic cascade of colorectal cancer. Several angiogenesis inhibitors have been evaluated in animal models and have shown efficacy, but challenges remain in using these drugs effectively in the clinical setting. Although several of these angiogenesis inhibitors are currently being evaluated in clinical trials, alone or in combination with cytotoxic chemotherapy, early results suggest that angiogenesis inhibitors alone, when used for advanced disease, have minimal activity. It is likely that this class of drugs will prove more efficacious when used either in the setting of minimal disease as agents that may promote tumor dormancy or in combination with other conventional forms of therapy. In addition, strategies such as metronomic therapy have been proposed whereby lower doses of cytotoxic chemotherapy, given more frequently, may act via an antiangiogenic mechanism [67,68]. Another challenge is identifying methods of assessing response to antiangiogenic therapy. To date, traditional methods of identifying response to treatment have not proven effective. Several investigators are working toward identifying circulating endothelial or tumor-associated factors that may be useful in following treatment. Novel imaging techniques are also being evaluated with enhanced CT and MRI, and newer modalities. Hepatic colorectal metastases provide an opportune setting in which to accomplish these challenges because the high incidence of disease and the ability to measure tumor with a variety of techniques lend themselves to evaluation of antiangiogenic therapy. PMID- 12735137 TI - A critical review of the major indicators of prognosis after resection of hepatic metastases from colorectal carcinoma. AB - Hepatic resections for metastatic colorectal cancer have dramatically increased, and there is clear evidence of the effectiveness of this type of surgery. Controversy, however, persists regarding appropriate patient selection, extent and timing of liver resection, and adjuvant or alternative therapeutic options. This article reviews the authors' experience with more than 600 hepatic resections and the relevant literature is discussed. The results underscore the importance of macroscopically and histologically complete tumor clearance, a so called "R0 resection." PMID- 12735138 TI - Arterial chemotherapy as adjuvant and palliative treatment of hepatic colorectal metastases: an update. AB - Regional hepatic chemotherapy with FUDR significantly improves local recurrence rates and may impact overall survival in patients with hepatic colorectal metastases. The results of prospective randomized trials confirm that careful patient selection, a thorough knowledge of intricate hepatic arterial anatomy, and an understanding of the pharmacokinetics and delivery of FUDR optimize treatment efficacy. A multimodality approach that includes adjuvant therapy in addition to cytoreductive surgery offers promise for the treatment of unresectable hepatic metastases. Because many tumors recur in extrahepatic sites, the addition of novel systemic agents such as CPT-11 may further reduce recurrences. Molecular analysis of the tumor may ultimately help select patients who are good candidates for regional chemotherapy. PMID- 12735140 TI - Incidence, distribution, and significance of hilar lymph node metastases in hepatic colorectal metastases. AB - For many surgeons, the presence of HLNM has been a formal contraindication to resection of LM from colorectal cancer. This attitude is based on the very poor survival seen in small subgroups of patients with HLNM who have been included in large-scale studies of patients with LM. The incidence of macroscopic HLNM in patients with LM has been reported at 1% to 12%. In the authors' experience, the rate of macroscopic HLNM is 7% and the incidence of macroscopic and microscopic HLNM is 19%. The reported 5-year survival rate of patients with resected HLNM is generally poor (12%), although this article reports a study with a 5-year overall survival rate of 27%. The authors do not recommend routine hilar lymph node biopsy and frozen section for all patients with LM undergoing resection; however, they do recommend a systematic palpation of hepatoduodenal lymph nodes with frozen section of suspicious lymphadenopathy prior to resection. In cases of proven HLNM, combined liver resection and lymphadenectomy could be considered in selected patients. This selection should be performed on an individual basis guided by the absence of important comorbid condition, the biology of the disease, and the surgeon's judgment that this is limited hilar lymphatic involvement without other sites of extrahepatic disease. Systematic routine en bloc lymphadenectomy currently has no prognostic value and no known therapeutic effects. PMID- 12735139 TI - Hepatic resection after down-staging of unresectable hepatic colorectal metastases. AB - Half of the patients with colorectal cancer present with liver metastases at some point in their illness. Surgical resection, which offers the only chance of long term survival, is an option in only 15% of patients at presentation. Novel chemotherapeutic treatments have enabled approximately 15% of those patients initially deemed inoperable to be down-staged to a point where surgery becomes an option. This aggressive approach, which requires close collaboration between surgeons and oncologists, results in a 40% 5-year survival rate, which is similar to that of patients operated without neoadjuvant chemotherapy. The hope for the future is the development of more effective chemotherapeutic regimens, which may allow more previously unresectable patients to benefit from curative surgery. This article reviews the literature and major areas of progress in treatments for unresectable colorectal cancer. PMID- 12735141 TI - Hepatic surgery for metastases from neuroendocrine tumors. AB - Cytoreductive therapy is effective in the management of metastatic neuroendocrine tumors to the liver, independent of their functioning status. In functioning tumors, clinical endocrinopathies are relieved in most patients and this response usually lasts for several months. Major morbidity and mortality are not greater than the average complication rate for resection for nonneuroendocrine metastatic tumors at major centers; therefore, surgical outcomes appear to justify operative intervention. Patients whose primary tumor can be controlled, whose metastases outside the liver are limited, and who have a reasonable performance status are candidates for resection. The authors' data support the previous statements. The current mortality rate of 1.2% and major morbidity rate of 15% clearly represent the success of the operative approach in such complex cases (54% of patients received a resection of at least one lobe) [9]. A symptomatic response in the 95% range with a median response of 45 months adds many months of symptom-free survival to the lives of most patients [9]. In the literature reviewed for this article, more than half of the patients also underwent a major hepatic resection and 40% of them had concurrent resection of the primary tumor. These data confirm that resection in selected patients is not more complicated or risky than resection for other metastatic tumors. Endocrinopathies have not increased anesthetic or operative risk in this population; however, these results are the product of managing these patients over time, becoming familiar with their clinical syndromes, and being active in the prevention of life-threatening endocrine complications (i.e., carcinoid crisis). The authors have learned over time that patients with valvular disease are not good candidates for surgery. These patients develop right-sided heart failure with an increase in the central venous pressure. This condition can result in massive hemorrhage during the liver resection because of the difficulty in controlling backbleeding from the hepatic veins [26]. Correction of valvular disease is warranted for safe liver resection. The authors' current policy is to rule out valvular disease in every patient with carcinoid tumors and repair the valves prior to hepatic resection when indicated [27]. This policy clearly has decreased the complication rate. Even though liver transplantation seems to be very attractive as a means of eradicating the disease, this has not been common in clinical practice because of the shortage of allografts, and the overall costs and complications of the procedure override its benefits, especially when compared with partial hepatectomy. Current methods to detect the spread of disease that were not readily available in the past, such as MRI and indium-111 pentetreotide (Octreoscan), may expand the applications of transplantation and allow for better selection of candidates. The option of transplantation is still open for improvement and is dependent on organ availability and better staging of the disease. Metastases from neuroendocrine tumors are hypervascular, favoring the application of MRI as the single imaging method; MRI not only evaluates the location and characteristics of the lesions but also determines the relationship with major vessels and bile ducts. Spiral CT scan has been used extensively in the past with acceptable results. Indium-111 pentetreotide functions on the base of somatostatin receptors present in these tumors, but its use has not been established definitely in the work-up of these patients. Perhaps the best use of indium-111 pentetreotide is in the evaluation of disease beyond the primary and liver locations, including bone metastases; its use therefore will likely affect the preoperative work-up of candidates for transplantation [28]. Once the patient has been deemed to have resectable disease by the preoperative work-up, several steps need to be completed prior to surgery to decrease the effect of specific endocrinopathies. For patients with symptoms related to carcinoid tumors, preoperative preparation with 150 to 500 micrograms of somatostatin decreases the chances of carcinoid crisis, which is manifested by hemodynamic instability [29]. The use of this medication intraoperatively should be kept in mind because a carcinoid crisis can occur despite anesthetic premedication. For islet cell tumors, treatment of underlying endocrinopathy has been initiated before referral for surgical treatment in most patients. Surgery is appropriate for patients with metastatic neuroendocrine tumors for the following two reasons: (1) many of them still have the primary tumor in place and resection should be undertaken to avoid acute complications and (2) the addition of adjunctive ablative therapies to surgical resection accomplishes the control of greater than or equal to 90% of the bulk of the tumor. If preoperative evaluation indicates that less than 90% of the tumor is treatable, surgical therapy is contraindicated. Last, even when complete resections are performed, the recurrence rate for these tumors is extremely high. In practical terms, patients with metastatic neuroendocrine tumors are seldom cured. The best hope physicians can offer these patients is an extended survival period with minimal endocrine symptoms and decreased requirements of somatostatin analogs. PMID- 12735142 TI - Complication, local recurrence, and survival rates after radiofrequency ablation for hepatic malignancies. AB - RFA is a technology that can be applied to unresectable hepatic malignancies without significant morbidity when patients and treatment approach are chosen carefully. The results of therapy provide patients with an improved chance for survival despite their unresectable disease. There are limitations of imaging to monitor treatment and treatment response. At this point, an intraoperative technique has greater treatment success than a percutaneous technique and this is likely associated with periprocedural imaging limitations with a percutaneous technique and the ability to more easily perform inflow vascular occlusion with an intraoperative approach. RFA is a potentially effective local tumor therapy for unresectable hepatic malignancies. Improved technology and concurrent application of RFA with other treatment modalities should continue to improve tumor response and patient survival for patients with unresectable hepatic malignancies. PMID- 12735143 TI - Chemotherapy for non-small-cell lung cancer. Part 2: Advanced disease. AB - The prognosis of patients with advanced non-small-cell lung cancer (NSCLC) remains poor. Systemic chemotherapy prolongs survival in this group of patients and palliates symptoms compared to best supportive care alone but more effective therapeutic strategies are needed. Novel agents that selectively target biological pathways of tumor growth offer hope of improving response and survival rates beyond what has been achieved with standard cytotoxic chemotherapy. Part 2 of this two-part article addresses the role of chemotherapy in locally advanced and advanced NSCLC, including the use of novel agents, considerations in elderly patients, and studies of second-line treatment. PMID- 12735144 TI - Current perspectives on anal cancer. AB - Anal cancer accounts for 1.5% of digestive system malignancies in the United States. In the past 30 years, substantial progress has been made in understanding the pathophysiology and treatment of the disease. Anal cancer was once believed to be caused by chronic local inflammation of the perianal area, and treatment was abdominoperineal resection. From epidemiologic and clinical studies, we now know that the development of anal cancer is associated with human papillomavirus infection and that the disease has a pathophysiology similar to that of cervical cancer. Less invasive treatments have also been developed, and the majority of patients with anal cancer can now be cured with preservation of the anal sphincter using concurrent external-beam radiation therapy and fluorouracil (5 FU)/mitomycin (Mutamycin) chemotherapy. Current areas under investigation include the incorporation of platinum agents into the chemotherapy regimen and the use of cytologic screening studies for high-risk populations. PMID- 12735145 TI - Clinical trials referral resource. Current clinical trials of imatinib mesylate. PMID- 12735146 TI - Vaccinations against infectious diseases in hematopoietic stem cell transplant recipients. AB - Blood and marrow transplantation, a curative treatment for a variety of serious diseases, induces a period of sustained immunosuppression predisposing recipients to opportunistic infections. Both for the protection of the individual transplant recipient and as a matter of public health policy, the US Centers for Disease Control and Prevention (CDC) has developed guidelines for the use of vaccination in the prevention of infectious disease following transplantation. This review examines the primary clinical research supporting vaccination policies in this target population. Widely accepted recommendations for transplant recipients based on scientific data are sparse, as few large studies have been conducted in this population. Anecdotal reports, expert advice, summaries, and limited series involving less than 50 patients using surrogate end points form the basis of the scientific literature, with the result being a wide variation in practice. Although based largely on inadequate scientific data, the CDC recommendations offer a pragmatic approach to the prevention of opportunistic disease in hematopoietic transplant recipients and serve as a useful starting point for standardization of practice while defining the direction of future studies in transplant recipients and other immunocompromised hosts. PMID- 12735147 TI - Opioids in cancer pain: common dosing errors. AB - Many individuals with advanced malignancy continue to suffer from pain and, consequently, impaired quality of life. The clinical scenarios in advanced cancer pain are complex, and successful management may require a more sophisticated and individualized approach than suggested by the World Health Organization guidelines. In patients referred to the Harry R. Horvitz Center for Palliative Medicine in Cleveland, numerous commonly occurring errors in opioid use have been noted. This article describes these errors and offers strategies with which to improve outcomes for patients suffering with cancer pain. PMID- 12735148 TI - High-risk head and neck cancers largely resist chemotherapy attack. PMID- 12735149 TI - Certification of added qualification in otology and neurotology. PMID- 12735150 TI - Old temporal bone fracture. PMID- 12735151 TI - Polyps arising in a concha bullosa of the middle turbinate. PMID- 12735152 TI - Nonoperative treatment of laryngeal granuloma. PMID- 12735153 TI - Electronystagmography: frequently overlooked significant findings. PMID- 12735155 TI - An endoscopic view of a wireless pH-monitoring capsule. PMID- 12735154 TI - Cranial neuropathy in neurosarcoidosis. PMID- 12735156 TI - Fibrous dysplasia of bone. PMID- 12735157 TI - Reimbursement basics. PMID- 12735158 TI - Tularemia of the head and neck: a possible sign of bioterrorism. AB - Recent bioterror attacks and other world events have focused the medical community's attention on agents that might be used in biological warfare. One of these potential biological weapons is Francisella tularensis, a gramnegative coccobacillus that is one of the most infectious bacteria known. F tularensis can cause severe, even fatal, systemic tularemia. Under normal circumstances, F tularensis is transmitted by infected ticks, insects, and other animals. As a weapon of terrorism, the bacterium would likely be disseminated as an aerosol and contracted by inhalation. Because many cases of tularemia are characterized by head and neck symptoms, otolaryngologists should be familiar with the diagnosis and management of this disease. In this article, we describe a case of zoonotic tularemia that manifested as a neck mass, and we review the pathophysiology, diagnosis, and treatment of tularemia. We also summarize what is known about its potential as a biological weapon. PMID- 12735159 TI - Temporal lobe encephalocele appearing as a lytic lesion of the skull base and pterygoid process. AB - Meningoencephalocele is an uncommon condition in which brain tissue, meninges, or both protrude through a defect in the anterior cranial fossa and into the ethmoid sinus or nasal cavity. Much less often, brain tissue, meninges, or both protrude through a defect in the middle cranial fossa and into the sphenoid sinus. We report an unusual case of a middle fossa encephalocele that appeared as a lytic lesion of the skull base. The patient was treated successfully via a unique endoscopic transpterygoid approach--that is, an endoscopic approach through the maxillary sinus and pterygopalatine fossa and into the pterygoid process. PMID- 12735160 TI - Pharmacologic manipulation of the labyrinth with novel and traditional agents delivered to the inner ear. AB - We describe the methodology and rationale behind the delivery of therapeutic medicines to the inner ear. The inner ear has long been impervious to pharmacologic manipulation. This is most likely the result of a protective mechanism called the blood-labyrinth barrier, whose function closely resembles that of the blood-brain barrier. This protective barrier impedes the clinician's ability to treat inner ear diseases with systemically administered medications. Since 1935, otolaryngologists have attempted to manipulate the inner ear with trans-tympanically injected medicines. Success has varied widely, but medicinal ablation of vestibular function can be achieved in this manner. Unfortunately, the auditory system is also at great risk from any medicine that is delivered to the inner ear via the middle ear. Over the past 10 years, significant improvements in drug delivery have allowed for more "titratable" treatment, which has reduced (but not eliminated) the risk of permanent hearing loss. In this article, we discuss both novel and time-tested methods of delivering medicines to the inner ear. We also review the classes of medications that alter inner ear function and the attendant risks of such treatments. PMID- 12735162 TI - Primary meningioma of the ethmoid sinus: a case report. AB - Meningioma is a well-recognized tumor of the central nervous system, but it rarely appears as a primary extracranial tumor of the paranasal sinuses. We report a case of a primary right anterior ethmoid meningioma that resembled a mucocele in its presentation. A primary meningioma can be differentiated from a secondary meningioma in three ways: (1) by observing an intact bony wall of the sinus on imaging or on inspection during surgery, (2) by noting the absence of a simultaneous intracranial meningioma on imaging or on inspection during surgery, and (3) by identifying a bulging of the sinus wall toward the cranium rather than in the opposite direction. PMID- 12735161 TI - The effects of a preoperative steroid/anesthetic injection on post-tonsillectomy pain. AB - We conducted a placebo-controlled, single-blind study to determine the efficacy of a local preoperative injection of a steroid/anesthetic combination in preventing post-tonsillectomy pain. We randomized 21 adults to receive either triamcinolone/bupivacaine on the left side and saline on the right or vice versa. Injections were administered in the area of the tonsillar pillars following intubation and prior to tonsillectomy. Based on the "generalized estimating equations" model of statistical analysis, we found no significant difference in the degree of postoperative pain between the active-treatment and control sides. PMID- 12735164 TI - Endoscopic management of inverted papillomas of the nose and paranasal sinuses. AB - We performed endoscopic resection on six patients who had inverted papillomas of the nasal cavity. Disease was completely cleared in four patients after a single procedure, while two others--both of whom had bilateral disease--required repeat surgery. At follow-up, which ranged from 6 months to 5 years, all patients were disease-free. We believe that endoscopic surgery is ideally suited for the treatment of appropriately selected patients who have limited disease and who are likely to adhere conscientiously to a regular follow-up program. PMID- 12735163 TI - Massive nasal polyposis in pediatric cystic fibrosis: report of an unusually severe case. AB - Nasal polyposis is common among children with cystic fibrosis. We report the case of a 14-year-old girl with cystic fibrosis who experienced severe facial disfigurement as a result of massive polyposis. Her polyps were removed surgically, and she was referred for cosmetic surgery. PMID- 12735165 TI - Extra-abdominal fibromatosis: a case report. AB - Head and neck fibromatosis is a rare disease with a variable clinical picture. We report the case of a 40-year-old woman who had a 1-week history of a painless mass overlying her right neck. Histology confirmed a diagnosis of extra-abdominal fibromatosis. Two unusual features of this case were the site of the lesion and its rapid growth. In this article, we discuss the clinical picture of head and neck fibromatosis, its etiology and pathology, and current opinion regarding its treatment. PMID- 12735167 TI - The evidence for universal care. PMID- 12735166 TI - Obstructive sleep apnea caused by bilateral vocal fold paralysis. AB - We describe the case of a woman who had been referred to us with a history of breathing difficulties and snoring and a suspicion of obstructive sleep apnea (OSA). Our investigation revealed that she did indeed have severe OSA in addition to undiagnosed bilateral vocal fold paralysis of unknown origin. Nocturnal polysomnography found that her apnea/hypopnea index was 120 and her minimum arterial oxygen saturation level was 63%. She was treated with laterofixation of the right vocal fold, and her OSA resolved immediately. During 10 years of follow up with nocturnal polysomnogrpahy, no recurrence of apnea or low oxygen saturation levels was noted. However, she did experience a recurrence of her snoring 4 years postoperatively, along with the onset of progressively worsening daytime fatigue. When these conditions persisted, we performed a repeat laterofixation of the same vocal fold. Following the repeat surgery, subjectively and objectively assessed results were good. PMID- 12735168 TI - Billing. Friendly bill, happier patient. PMID- 12735169 TI - Readiness. Just in case. PMID- 12735170 TI - Patient safety. It's an infection. Is it a lawsuit? PMID- 12735171 TI - EMRs. Information, M.D. PMID- 12735172 TI - EMRs. What's the government's role? PMID- 12735173 TI - Design-build. From the ground up. PMID- 12735174 TI - Temp jobs tempt docs. PMID- 12735175 TI - Employee satisfaction. Room with a view. PMID- 12735176 TI - Telemedicine. Dial-up care. PMID- 12735177 TI - Confidentially speaking. PMID- 12735178 TI - Consumer confusion. AB - Predictions of the rise of a take-charge health care consumer have proved premature. Opinion polls and data paint a confusing portrait of a consumer who wants quality care at discount prices, but who doesn't know how the system works, what it costs or what his or her role is. When will this super consumer emerge, and will the industry change as a result? PMID- 12735179 TI - MDs + IT. Facilitating physician adoption. AB - This month's gatefold examines how physicians view and use information technology, the reasons for physician resistance to IT and strategies that hospital leaders use to overcome that resistance and incorporate IT into their organizations. PMID- 12735180 TI - Inoculating for smallpox. AB - In California, one hospital decided the time was right to inoculate staff volunteers with the smallpox vaccine. Weighing the same pros and cons--civic responsibility, health risk, cost and reluctance of staff--other hospitals opted out of inoculations, at least for now. What led these organizations to such divergent decisions? PMID- 12735182 TI - For the benefit of the community. AB - As the head of Phoenix's John C. Lincoln Health Network, Dan Coleman has helped revitalize the organization's community services program. For its wide-ranging efforts, including real estate development, a food bank, family support programs, a children's dental clinic and a day care center, John C. Lincoln won this year's Foster G. McGaw Award. PMID- 12735183 TI - The art of listening. PMID- 12735184 TI - Renegotiating the social contract. PMID- 12735185 TI - Executive insights. John J. Gantner, treasurer, Robert Wood Johnson University Hospital, New Brunswick, New Jersey. AB - Sometimes a CFO has to make a leap of faith that he or she is making the right decisions, because those decisions can involve intangibles that reach far beyond financial data. According to John J. Gantner, treasurer of Robert Wood Johnson University Hospital in New Brunswick, N.J., if your decisions are guided by what is best for patient care, they will likely benefit the hospital over the long term. Gantner's hospital has taken a leadership role in adopting standards relating to patient safety, fiscal accountability, and ethics. Gantner spoke with HFM about what has guided some of the recent major initiatives undertaken at his hospital. PMID- 12735187 TI - Drug card setback highlights need for cost relief. AB - Under Bush's prescription card program, Medicare beneficiaries could access the mechanisms used by the privately insured population to lower prescription drug costs and use the leverage of the senior market share to negotiate deeper discounts from drug manufacturers. PMID- 12735186 TI - Containing revenue-cycle costs. AB - Healthcare organizations can achieve revenue-cycle improvement while maintaining optimum staffing levels by taking a three-step approach-developing a plan, measuring costs, and using benchmarks to measure performance. Planned cost reductions can be achieved without a negative impact on the organization. Cost reduction strategies should look at purchased services, staffing, and the cost-to collect ratio. Healthcare organizations should reach target performance levels before implementing a cost-reduction strategy. PMID- 12735188 TI - Compliance. What's around the corner? AB - Compliance officers and financial managers should forge an alliance to ensure their organization's ethics and integrity. Many financial and compliance issues stem from clinical research. Billing for experimental devices must follow specific rules, especially when payment is from Medicare. The charge description master should receive detailed attention to improve efficiency and net revenues. Violations of protected health information can have serious financial implications. PMID- 12735189 TI - What price is right? AB - Pricing is an effective strategy for increasing hospital revenue. Usually between 10 and 25 percent of a hospital's business is charge-related. Hospital executives should consider price-driven payment, such as that affected by carve-out arrangements. Critical pricing issues must be thoroughly understood to perform an effective pricing study. PMID- 12735190 TI - Best practices for preempting payment denials. AB - Limiting inpatient claims denials requires a combination of data, processes, and people, including: Systems that support collecting correct payer data; Creative processes to prevent denials in high-denial areas; Compliance with payer protocols; Enhanced front-end staffing. PMID- 12735192 TI - Coaching managers. PMID- 12735191 TI - Preparing for asset retirement. AB - Statement of Financial Accounting Standards (SFAS) No. 143 requires organizations to recognize a liability for an asset retirement obligation when it is incurred- even if that occurs far in advance of the asset's planned retirement. For example, organizations must recognize future costs associated with medical equipment disposal that carries hazardous material legal obligations. PMID- 12735193 TI - Why does an IT investment fail to deliver returns? PMID- 12735194 TI - MSA market factors determine hospitals' appropriate level of charges. PMID- 12735196 TI - Hamstring tendon harvesting. Reviewing anatomic relationships and avoiding pitfalls. AB - The gracilis and semitendinosus tendons remain popular grafts for many reconstructive procedures, including anterior cruciate ligament reconstruction. Surgeons who use other grafts for ACL reconstruction should have the hamstring tendon techniques in their arsenal in the event other grafts are unavailable or undesirable. Detailed knowledge of the anatomy of the medial side of the knee is paramount to avoid potential pitfalls and ensure a successful tendon harvest. PMID- 12735197 TI - ACL reconstruction with hamstring tendon. AB - The technique of quadrupled semitendinosus autograft for ACL reconstruction using the EndoButton for femoral fixation has been described. Dr. Rosenberg, this article's senior author, has used this for over 10 years with no known instance of fixation failure at the femur or tibia. This technique using QST reconstruction has little morbidity, low reoperation rate, and excellent clinical results. PMID- 12735198 TI - Scientific justification and technique for anterior cruciate ligament reconstruction using autogenous and allogeneic soft-tissue grafts. AB - The DLSTG is the strongest and stiffest autogenous graft source available for reconstruction of the torn anterior cruciate ligament. Harvest morbidity is low compared with other autogenous graft sources, such as the patellar bone-tendon bone graft. Soft-tissue allografts provide an excellent alternative for patients requiring revision surgery or for patients who want to avoid any morbidity associated with autogenous graft harvest. Successful use of any soft-tissue graft source, however, relies on precise placement of the tibial and femoral tunnels to prevent roof and PCL impingement and to restore tensile behavior in the graft tissue similar to the native ACL. The use of high-strength, high-stiffness fixation devices that secure the graft at the end of the tunnel promote tendon tunnel healing, restore stability without high graft tensioning, and allow safe, aggressive rehabilitation. The Bone Mulch Screw/WasherLoc screw system provides the surgeon with a consistent, reproducible technique that restores stability and function to the ACL-deficient knee using a soft-tissue graft in both males and females. PMID- 12735199 TI - Anterior cruciate ligament reconstruction utilizing central quadriceps free tendon. AB - CQFT ACL reconstruction yields a stable, highly functional knee with little associated morbidity. Patient satisfaction has been very high, and objective follow-up data collected thus far is very encouraging. PMID- 12735200 TI - Anterior cruciate ligament reconstruction with allograft tendons. AB - Allograft tissue allows reconstruction of the ACL without the donor site morbidity that can be caused by autograft harvesting. Patients who must kneel as a part of their occupation or chosen sport are particularly good candidates for allograft reconstruction. Patients over 45 years of age and those requiring revision ACL surgery can also benefit from the use and availability of allograft tendons. In some cases, patients or surgeons may opt for allograft tendons to maximize the result or morbidity ratio. Despite advances in cadaver screening and graft preparation, there remain risks of disease transmission and joint infection after allograft implantation. Detailed explanation and informed consent is vitally important in cases in which allograft tissue is used. PMID- 12735202 TI - Special considerations. Anterior cruciate ligament reconstruction in the skeletally immature. AB - ACL injury in children is being recognized with greater frequency because of improved diagnostic techniques as well as heightened awareness of the condition. Unfortunately, the diagnosis is still missed because the attitude persists that children do not suffer ligament injuries. Hemarthrosis must be considered an indication of a significant intra-articular injury. During the past decade, ACL reconstruction has evolved to a reproducible technique with low morbidity. Aggressive rehabilitation programs allow accelerated return to activity while allowing the biology of graft maturation to progress. The basic principle of diagnosis and the treatment goals in the skeletally immature patient are the same as those in the adult patient. The diagnosis approach to ACL injury in the scholastic-age patient, however, must also include evaluation of the patient's skeletal maturity because it plays a major role in treatment decisions. Maturity is evaluated on the basis of the patient's chronologic age; various physiologic factors, such as family height, patient's projected height, and estimation of sexual development; and radiographic findings in the knee, pelvis (Risser sign), or hand and wrist (bone-age study). Because of the special characteristics of the skeletally immature patient, the orthopedic surgeon must act as "knee counselor" by attempting to identify at-risk patients, particularly those who abuse their knees for any of a variety of reasons. The nonoperative treatment principles are the same as those in an adult. Consideration of surgical treatment must take into account assessment of skeletal maturity. If questions remain about the status of the femoral and tibial physes, polytomography or MRI is used to assess the extent of physeal closure. The surgical reconstruction used reflects the patient's skeletal maturity. As the skeletal maturity threshold is reached, transphyseal reconstructions may be done with diminished reservation about causing sequelae of physeal arrest. PMID- 12735201 TI - Anterior cruciate ligament reconstruction using semitendinosus and gracilis tendons, bone patellar tendon, or quadriceps tendon-graft with press-fit fixation without hardware. A new and innovative procedure. AB - BONE--PATELLAR TENDON: The "no hardware" technique for ACL reconstruction is a new method that offers many advantages and is straightforward to perform. Its main innovative feature is that it does not require bone-block harvesting from the patella. This reduces donor site morbidity and prevents patellar fractures. The bone tunnels are made using tube harvesters and compaction drilling. This minimizes trauma and obviates the risk of bone necrosis. The articular entrance of the tibial tunnel is completely occupied by the grafts. This prevents a windshield-wiper effect and synovial fluid ingress into the tunnel, and enhances graft incorporation. The fact that no hardware is used with both patellar tendon or hamstring grafts significantly reduces the overall cost of the operation and facilitates revision surgery. The quadriceps tendon is also a very good graft. It is thick and has good biomechanical properties and low donor site morbidity. Its disadvantages are: weakness of quadriceps after the operation, an unsightly scar, and some difficulty in graft harvesting [58]. Also, postoperative MRI is not fraught with the problem of metal artifacts. It is difficult to decide which of the methods currently available for ACL reconstruction is the best because most of them give satisfactory results. In the future, assessments of knee ligament reconstruction techniques should look at long-term stability combined with low complication rates. Ease of revision surgery and low cost should also be taken into consideration, given the large annual volume of knee ligament reconstructions (50,000 in the United States alone) [59]. We believe that our technique addresses most of these issues, and that it constitutes a useful alternative method for ACL reconstruction. SEMITENDINOSUS--GRACILIS: This technique, which was used with 915 patients from June 1998 to February 2002, shows a particularly low rate of postoperative morbidity. The reason is probably to be found in the "waterproofing" of the bone tunnels, which lead to less postoperative bleeding and swelling. No drains were used. Rehabilitation follows the same protocol as used for the reconstruction using patellar tendon grafts (accelerated/functional). As expected, there was no widening of the femoral tunnels and little widening of the tibial tunnels. Interestingly, tibial tunnel enlargement was significantly less in a nonaccelarated rehabilitation group than in the accelerated group [60] without affecting stability. The measured internal torque of the hamstrings, as well as their flexion force, already had returned to normal 12 months postoperatively. In a prospective randomized (unpublished) study comparing this technique with ACL reconstruction with BPT grafts with medial or lateral third with only one bone plug (from the tibial tuberosity, see technique described above), we found no significant difference between both groups in subjective scores, stability, KT-1000 values, Tegner activity score, and IKDC at 1-year follow-up. Only the results of kneeling and knee walking testing were significantly better in the hamstring group [61]. In summary, the advantages of this presented technique are: (1) the knot of the graft is close proximally to the anatomic site of the insertion of the ACL, thus avoiding the Bungee effect.; (2) the press-fit tunnel fixation prevents synovial fluid entering the bone tunnels, windshield-wiper effect, and longitudinal motion within the tunnel; the intensive contact between the bony wall of the tunnel and graft collagen over a long distance without any suture material results in quick and complete graft incorporation; and (3) no fixation material means no hardware problems, facilitates revision surgery, and lowers overall costs. PMID- 12735203 TI - Revision anterior cruciate ligament reconstruction. AB - Revision ACL surgery is indicated in patients who present with pathologic anterior laxity on clinical examination that reproduces their symptoms of instability during activities of daily living or athletic activities. The goals of the revision ACL surgery are to stabilize the knee, prevent further injury to the articular cartilage and menisci, and maximize the patient's function. Successful revision ACL surgery requires a thorough preoperative evaluation, including a detailed history, physical examination, and radiographic evaluation. Preoperative planning begins with a determination of the mechanisms of failure for the initial ACL reconstruction. Often a primary, as well as secondary cause, for failure can be identified. The determination of the cause of failure is the first step in a carefully-constructed treatment plan, which includes consideration of skin incisions to be used, method of graft removal, hardware removal, the need for a staged procedure or concomitant surgery, graft material selection, tunnel placement, graft fixation, and postoperative rehabilitation protocol. Despite the most meticulous planning, unanticipated findings may be encountered in the operating room, and the preoperative plan should have enough flexibility to accommodate these developments. Finally, it is crucial to counsel the patient preoperatively to limit his or her expectations regarding their surgical outcome. Given the complexity of revision ACL reconstruction, patient expectations must be adjusted to realistically match the potential for success. With proper planning, attention to detail, and appropriate patient expectations, revision ACL surgery can result in a beneficial and satisfying patient outcome. PMID- 12735204 TI - Complications in anterior cruciate ligament surgery. AB - There are many potential problems associated with ACL surgery; however, careful attention to detail can prevent problems from occurring or provide corrective options in the event they do occur. Multiple backup options are not only desirable but also essential to a successful outcome. By following these guidelines, ACL surgery can have good long-term results and return the recreational or higher-level athlete to sports participation. PMID- 12735205 TI - Recent advances in the rehabilitation of isolated and combined anterior cruciate ligament injuries. AB - The rehabilitation process begins immediately following ACL injury, with emphasis on reducing swelling and inflammation; improving motion; regaining quadriceps control; allowing immediate weight-bearing; and restoring full passive knee extension and, gradually, flexion. The goal of preoperative rehabilitation is to prepare the patient mentally and physically for surgery. Once the ACL surgery is performed, it is important to alter the rehab program based on the type of graft used and any concomitant procedures performed. This will aid in preventing several postoperative complications, such as loss of motion, patellofemoral pain, graft failure, and muscular weakness. The goal of this article has been to provide an overview of the application and the scientific basis for formulating a rehabilitation protocol following ACL surgery. For an athlete to return to competition, it is imperative that he or she regain muscular strength and neuromuscular control in their injured leg while maintaining static stability. In the past, rehabilitation programs attempted to prepare the athlete for return to sports by using resistance exercise alone. Current rehabilitation programs focus not only on strengthening exercises, but also on proprioceptive and neuromuscular control drills in order to provide a neurologic stimulus so that the athlete can regain the dynamic stability needed in athletic competition. We believe that it is important to use this approach not only possible causes that might predispose the individual to future injury. PMID- 12735206 TI - Approach to meniscal tears in anterior cruciate ligament reconstruction. AB - It is generally accepted that meniscus repair in a knee with persistent anterior cruciate ligament (ACL) insufficiency leads to a higher retear rate, and that meniscal repair in conjunction with ACL reconstruction is associated with a higher rate of meniscal healing compared with patients who do not undergo ACL reconstruction. In this article, we will review the background of thses concepts and present treatment strtegies for meniscal tears in ACL-insufficient knees, with particular attention to meniscal repair and meniscal allograft transplanatation. PMID- 12735207 TI - Approach to cartilage injury in the anterior cruciate ligament-deficient knee. AB - The treatment of articular cartilage lesions remains one of the great challenges facing orthopedic surgeons today. The technique of chondrocyte transplantation has opened the door for the application of biologic solutions to difficult problems. These techniques will prove the keystone of further advances into biologic joint repair and replacement. Enthusiasm, however, must be tempered by the numerous gaps in cartilage science and the overwhelming need for further long term data to demonstrate the efficacy of these techniques in thwarting the presumed eventual progression of these lesions toward osteoarthritis. The status of the articular cartilage is of paramount importance in ACL decision-making. Every effort must be made to protect the existing hyaline articular cartilage during ACL reconstruction. Though current cartilage repair techniques are in their infancy, they remain stepping-stones to future developments. It is hoped that we will one day be able to regenerate normal hyaline cartilage without great morbidity. At present, the ACL surgeon must accept techniques that diminish symptoms and do not burn bridges to future advances. The orthopedic surgeon must increase his knowledge of the basic science of articular cartilage in order to best choose from the various cartilage treatments that evolve. PMID- 12735208 TI - Arthritis and osteotomies in anterior cruciate ligament reconstruction. AB - Unfortunately, young patients with active life-styles who present with an ACL deficient knee and early evidence of arthritis remain a very difficult population to treat. Arthroscopic ACL reconstruction is a proven and effective way to treat patients with anterior knee instability [2]. In addition, numerous authors have demonstrated that knee osteotomies are effective in addressing isolated medial or lateral compartment degenerative disease, even in the presence of anterior knee instability [7, 9, 22, 43]. A comprehensive evaluation of pain, alignment, and stability are necessary in order to formulate the best treatment plan for each patient. Patients with combined chronic instability secondary to ACL insufficiency and unicompartmental arthritis will generally benefit from arthroscopic debridement, ACL reconstruction, knee osteotomy, or any combination thereof. Expectations of patients should be managed according to their underlying pathology, and this is a critical component for the success of the surgery. Most patients will get some degree of symptomatic relief and functional improvement. A return to competitive or high-level sports is an unrealistic goal, however, and should be discouraged. PMID- 12735209 TI - Expert panel on anterior cruciate ligament case studies. AB - Anterior cruciate ligament (ACL) reconstructions cannot, and should not, be considered "recipe-driven" or "cookie-cutter" operations. There are multiple variables that must be considered both pre- and intraoperatively in order to execute a successful procedure. The following case studies present a number of challenging situations, many of which do not have a straightforward, or only one, solution. The goal is to provide some ral-life clinical situations in which the reader can "observe" an expert panel of ACL surgeons working through the problems -offering solutions based on science, experience, and intuition. PMID- 12735210 TI - Immunogenicity of erythropoietin and other growth factors. AB - Erythropoietin (EPO) is a 165 amino-acid sequence glycoprotein which plays an important role in maintaining regular generation of erythrocytes, Jacobs et al. describe the cloning of the human EPO and recombinant EPO has been introduced for the treatment of anemia in patients with renal diseases. The extensive utilization of EPO can induce the production of neutralizing EPO antibodies, which have been proved in patients with the non-infectious form of pure red cell aplasia (NI-PRCA), an autoimmune disease characterized by a sudden inhibition of erythrocyte maturation and production. In this review, the literature concerning the molecular structure and the genetic profile of EPO as well as the relationship between neutralizing EPO antibodies and NI-PRCA have been analyzed. PMID- 12735211 TI - Anemia in renal insufficiency. AB - In renal failure, severe anemia and associated fatigue, cognitive and sexual dysfunction have a significant impact on the patient's quality of life. Anemia has also been identified as an important etiologic factor in the development of left ventricular hypertrophy. The major cause of anemia in presence of a reduction of glomerular filtration rate is an inadequate production of a glycoprotein hormone, the erythropoietin (EPO). EPO is the primary regulator of the growth and survival of erythroid progenitor. The introduction of recombinant human erythropoietin (rHuEPO) has revolutionized the treatment of anemia in chronic renal failure. The vast majority of patients respond very well to treatment, but 5-10% of patients show some resistance to EPO, the most common cause of which is iron deficiency. Several studies are recently commenced to investigate the effects of preventing renal anemia ever developing. The target of hemoglobin concentration in pre-dialysis and dialysis patients are object of continuous re-examinations. PMID- 12735212 TI - Erythropoietin and chronic lymphocytic leukemia. AB - Anemia is a frequent clinical feature with adverse prognostic effects in patients with chronic lymphocytic leukemia (CLL). It may complicate CLL at any time during the course of the disease. Different factors concur to the occurrence of anemia in CLL, as in other lymphoproliferative diseases: leukemic bone marrow infiltration, the myelosuppressive effect of chemotherapy and inhibiting cytokines, autoimmune phenomena, hypersplenism, a poor nutritional status that leads to folic acid, vitamin B12 and iron deficiency. In addition, a defective endogenous erythropoietin (EPO) production has also been described in patients with lymphoproliferative diseases. The severity of anemia, which may be worsened by an impaired cardiopulmonary function, may profoundly compromise the patients' quality of life and, indirectly, the outcome of cancer bearing patients. Several Authors have reported the clinical activity of recombinant human (rHu)EPO in anemic patients with lymphoproliferative diseases, including CLL. Low serum EPO levels at baseline and EPO levels inappropriately low for the degree of anemia help to identify patients who are likely to respond to EPO. A clear dose dependent response to EPO has been reported by different Authors and it has been suggested that 5,000 IU should be considered as an appropriate initial dose for the majority of patients. rHuEPO represents a potentially effective and safe therapy for the management of anemia associated with lymphoproliferative diseases. The reduction of red blood cell transfusion requirement, the improvement of quality of life through the remission of fatigue-related anemia are two important results that should be considered in the management of patients with CLL. In prospect, the availability of new rHuEPO molecules with a more prolonged half-life may open new therapeutic avenues. PMID- 12735214 TI - Upregulation of erythroblast apoptosis by malignant plasma cells: a new pathogenetic mechanism of anemia in multiple myeloma. AB - Anemia of variable severity occurs in more than two-thirds of patients with multiple myeloma (MM). Besides the altered cytokine network, chronic erythropoietn deficiency, blood loss and hemolysis, we have shown that deregulated myeloma cell apoptosis contributes to progressive destruction of the erythroid matrix by inducing erythroblast cytotoxicity. To exert this effect, highly malignant plasma cells overexpress both Fas-L and tumor necrosis factor related apoptosis-inducing ligand (TRAIL), which efficiently trigger the death of immature erythroblasts. However, this Fas-L/TRAIL-based anemia occurs in particular in patients with severely progressive MM, thus suggesting that these apoptogen receptors may characterize a peculiar cytotoxic-apoptogenic phenotype of malignancy. Immunophenotyping of myeloma cells could help to identify patients with a higher risk of erythropoiesis exhaustion. PMID- 12735213 TI - The role of recombinant human erythropoietin alpha in the treatment of chronic anemia in multiple myeloma. AB - Chronic anemia of variable severity occurs in more than two-thirds of patients with multiple myeloma (MM) as a consequence of the B cell malignancy. Its pathogenesis is multifactorial. Besides the altered inflammatory cytokine network, other events are held responsible, namely persistent defect of erythropoietin due to the kidney failure, shortening of red cell survival, accumulation of the serum monoclonal component and platelet dysfunction. Our recent studies have demonstrated that excessive erythroblast apoptosis promoted by myeloma cells drives the appearance of anemia, in particular in patients with severely progressive disease. A number of clinical trials have provided evidence for the effectiveness of recombinant human erythropoietin (rHuEPO-alpha: epoetin alpha) in improving the deregulated erythropoiesis in MM, since it acts as a major erythroid growth factor by exerting a specific anti-apoptotic effect. In the majority of these studies, the long-term treatment of MM-associated anemia with rHuEPO-alpha induced a significant improvement of erythropoiesis, as shown by a stable increase of hemoglobin values (> or = 2g/dL) and reduction of transfusion requirements. In a recent trial which included both a double-blind and an open-label phase, we have documented that rHuEPO-alpha induces a stable improvement of anemia in more than 75% of patients and a significant decrease of fatigue, with an overall recovery of the quality of life. Patients receiving a placebo also achieved similar results in the open-label phase, when they were switched to rHuEPO-alpha. PMID- 12735215 TI - NNRTI hepatotoxicity: efavirenz versus nevirapine. PMID- 12735216 TI - Preventing prematurity. PMID- 12735218 TI - Genes linked to severe lupus. PMID- 12735217 TI - Is an ephedra ban coming? PMID- 12735219 TI - Update: cervical cancer screening. PMID- 12735220 TI - Obesity in pregnancy. What every nurse needs to know. PMID- 12735221 TI - Amniotic fluid embolism. Exploring this rare but typically fatal condition. PMID- 12735222 TI - Self-administered pain medications. A practical approach in an OB/GYN setting. PMID- 12735223 TI - Deconstructing DNA. Understanding genetic implications on nursing care. PMID- 12735224 TI - Cultural diversity in the workplace. How the meaning of work can affect the way we work. PMID- 12735226 TI - Metabolic profiling and biomarker discovery. PMID- 12735225 TI - Laboring lessons. Everyday encounters with the beauty of life. PMID- 12735227 TI - Approaches in anticoagulation: rationales for target positioning. AB - Heparin and warfarin are the most widely used anticoagulants for the prophylaxis and treatment of thrombus-based diseases. These anticoagulants, however, have well-known clinical limitations, such as a slow onset of action and a narrow therapeutic window. An ideal small-molecule non-peptide inhibitor should have an immediate onset of action, oral bioavailability and an improved therapeutic action and side-effect profile, compared to established therapies. In this review, the current concepts and hypotheses of the numerous anticoagulant approaches are analyzed and evaluated, with emphasis on animal models, genetic disorders and compound profiling. Selected factors of the coagulation cascade and modulators of endogenous fibrinolysis are examined to determine if they represent promising drug targets in antithrombotic therapy. PMID- 12735228 TI - Factor Xa inhibitors: today and beyond. AB - Serine proteases play an important role in thrombogenesis, the process that leads to blood clotting and conditions such as heart attack, stroke and other cardiovascular disorders. In the coagulation network, the activation of various serine proteases facilitates the formation of the serine protease Factor Xa, which plays a central role in the process of coagulation and platelet activation. Factor Xa is an essential component of the prothrombinase complex, from which thrombin is formed, which then directly leads to fibrin clot formation. Thus, the inhibition of Factor Xa and its generation is an important strategy in the development of new antithrombotic drugs. PMID- 12735230 TI - Therapeutic implications of cholesteryl ester transfer protein inhibitors in hyperlipidemia and low high-density lipoprotein-cholesterolemia. AB - Low levels of high-density lipoprotein cholesterol (HDL-C) in plasma are an independent coronary risk factor. Therapies that lower cholesteryl ester transfer protein (CETP) have preventative effects on aortic atherosclerosis in cholesterol fed rabbits. CETP inhibitors are a new class of compounds that can increase HDL-C levels by up to 70%, according to data from phase I and II clinical trials. CETP inhibitors are therefore likely to be beneficial in patients with moderate hypercholesterolemia and HDL-C levels lower than 40 mg/dl. CETP inhibitors should, however, be viewed with caution as their effects on triglyceride metabolism are currently unknown. PMID- 12735229 TI - Targeting sources of superoxide and increasing nitric oxide bioavailability in hypertension. AB - Overproduction of oxygen free radicals, which is mainly mediated by superoxide, occurs in human hypertension and a wide variety of animal models. There are several important enzymatic sources of superoxide production, including NADPH oxidase, xanthine oxidase and uncoupled nitric oxide synthase. Superoxide levels are also controlled through endogenous antioxidant systems and superoxide dismutase is the primary antioxidant in the vascular system. Strategies have therefore focused on combating hypertension and vascular disease through the inhibition of superoxide-generating enzymes, and scavenging superoxide. While results from animal studies are promising, no consensus has been reached on identifying a drug target for the reliable and effective treatment of oxidative stress in hypertension. PMID- 12735231 TI - QT interval prolongation and cardiac risk assessment for novel drugs. AB - Fatal cardiac arrhythmias, known as torsades de pointes, can occur with a wide variety of medicinal drugs and are associated with prolongation of the QT interval. This review critically evaluates the major strategies for assessing QT prolongation risk: ion channel studies, in vitro cardiac electrophysiology, and in vivo cardiac electrophysiology and hemodynamics. Disease- or drug-induced QT prolongation is mainly associated with reduced amplitude of the repolarizing outward K+ current in myocardial cells, particularly those carried by the human ether-a-go-go-related gene (HERG) channel. Thus, measuring HERG currents using patch-clamp technology and cloned HERG channels represents a first approach for evaluating adverse effects of drugs on ion channel function, under physiological conditions. Evaluation of changes in transmembrane action potential in isolated rabbit or dog Purkinje fibers reflects mixed ion channel blocking properties of the test substance and therefore permits a greater understanding of the mechanisms underlying the genesis of arrhythmias. Both HERG channel and Purkinje fiber procedures are clinically predictive, however, no in vitro technique can fully reproduce the in vivo situation. Therefore, both in vitro and in vivo approaches should be employed to maximize the chances of an accurate assessment of risk in an area where prolonged QT can result in death. PMID- 12735232 TI - Recent advances in the diagnosis and treatment of deep vein thrombosis: a regional consensus. AB - Considerable progress has been made in the understanding of the risk factors for venous thromboembolism (VTE). The clinical applications of molecular techniques have allowed identification of important inherited, yet not uncommon, risk factors for VTE, such as mutations that cause Factor V Leiden and prothrombin G20210A. However, advances in our understanding have raised several questions regarding the need for, and duration of anticoagulation. At the end of the treatment period, low molecular weight heparins have become the drugs of choice and standard-of-care for VTE. In this review, cost effective diagnostic approaches for patients with suspected deep vein thrombosis, and recommended treatment options using evidence-based approaches, are described. PMID- 12735234 TI - SLV-306. Solvay. AB - SLV-306 is an orally active mixed neutral endopeptidase/endothelin converting enzyme inhibitor under development by Solvay SA for the potential treatment of essential hypertension and congestive heart failure. The compound is currently undergoing phase II clinical trials in Belgium. PMID- 12735233 TI - Tezosentan. Actelion/Genentech. AB - Tezosentan is an endothelin receptor antagonist in development by Actelion Ltd and Genentech Inc for the potential treatment of cardiovascular disorders, including acute heart failure (AHF) and congestive heart failure. Actelion has conducted a phase III study of tezosentan in AHF patients, however, following disappointing trial results a new phase III study in AHF was planned using lower doses of the compound. By January 2003, Actelion anticipated that the first results of a two-year phase III study evaluating mortality/morbidity would be available by early 2005. PMID- 12735235 TI - MLN-519. Millennium/PAION. AB - Millennium Pharmaceuticals Inc (formerly LeukoSite Inc) and PAION GmbH are developing MLN-519, a ubiquitin/proteasome enzyme inhibitor, for the potential treatment of inflammatory diseases and stroke. MLN-519 is currently undergoing phase I clinical trials in acute stroke and myocardial infarction, and is poised to enter phase II trials. PMID- 12735236 TI - BO-653. Chugai. AB - BO-653 is an antioxidant under development by Chugai for the potential treatment of atherosclerosis and the prevention of restenosis. By November 2001, BO-653 was in phase II trials for restenosis in post-percutaneous transluminal coronary angioplasty in the US, and by April 2002, the compound was in phase I trials for the same indication in Japan. PMID- 12735237 TI - Eflucimibe. Pierre Fabre/Eli Lilly. AB - Eflucimibe is an acyl coenzyme A:cholesterol acyltransferase inhibitor under development by Pierre Fabre SA and Eli Lilly & Co for the potential treatment of hypercholesterolemia and atherosclerosis. Phase II clinical trials commenced during 2002. PMID- 12735238 TI - RSD-1235. Cardiome. AB - RSD-1235 is a novel, mixed ion channel antagonist in development by Cardiome Pharma for the potential treatment of heart arrhythmia. By September 2002, phase II trials had been completed and phase III studies were due to commence in 2003. PMID- 12735239 TI - Ectomycorrhizas involving sebacinoid mycobionts. AB - Nuclear ribosomal DNA sequences characteristic of Sebacinaceae were detected by direct amplification of DNA from field collected ectomycorrhizal samples. A study was undertaken to confirm the formation of ectomycorrhizas by Sebacinaceae and to characterise representative samples genetically, morphologically and ultrastructurally. The investigated sebacinoid mycorrhizas were sufficiently characteristic to be identified morphologically. They are distinguished by a characteristic, clampless, hydrophilic extramatrical mycelium, which is very variable in diameter and in wall thickness, and by the presence of distinctive, y shaped, inflated multibranchings. Differences in the mantle structure clearly discriminate the two investigated sebacinoid mycorrhizal types. Comparison of the D1/D2 domains of the nuclear large subunit pseudogene and the ITS1 and ITS2 regions identifies the fungal partner of one of the mycorrhizal samples as Sebacina incrustans. Ultrastructural investigations of the ectomycorrhizas show a doliporus/parenthesome architecture consistent with that of the Sebacinaceae. Recently published sequence data obtained from sebacinoid mycorrhizas are compared to our sequences and the complex trophic relationships in the Sebacinaceae are discussed. Observations on ectomycorrhizas and basidiomes suggest that species of Sebacinaceae are fairly common mycobionts in various ectomycorrhizal plant communities. PMID- 12735240 TI - Phylogenetic analysis of Puccinia distincta and P. lagenophorae, two closely related rust fungi causing epidemics on Asteraceae in Europe. AB - Phylogenetic analyses of the ITS1-5.8S-ITS2 region of the ribosomal RNA gene cluster were carried out with two short-cycled (aecial/telial) European rusts on Asteraceae, Puccinia distincta causing the current pan-European epidemic on Bellis perennis, and P. lagenophorae causing a similar disease on Senecio spp., as well as the macrocyclic P. obscura which alternates between B. perennis (pycnial and aecial host) and Luzula spp. (main host). All three species formed a well-resolved cluster when compared with the ITS sequences of a range of other rust fungi, using both parsimony and distance methods. The sequences of P. distincta and P. lagenophorae differed from each other at three positions whereas P. obscura differed from P. distincta at 37 points. Together with consistent morphological and epidemiological differences across Europe, these data support the recognition of P. distincta as a separate species from P. lagenophorae. Both may be derived from P. obscura, although the precise evolutionary history remains obscure. PMID- 12735241 TI - Nuclear and mitochondrial rDNA variability in Crinipellis perniciosa from different geographic origins and hosts. AB - Genetic variability in Crinipellis perniciosa, the causal organism of witches' broom disease in Theobroma cacao, was determined in strains originating from T. cacao and other susceptible host species Heteropterys acutifolia and Solanum lycocarpum in Brazil, in order to clarify host specificity and geographical variability. RFLP analysis of the ribosomal DNA ITS regions (rDNA ITS), and the mitochondrial DNA small subunit ribosomal DNA gene (mtDNA SSU rDNA) did not reveal any genetic variability in 120 tested strains, possibly serving only as species level markers. Genetic variability was observed in the ribosomal DNA IGS spacer region, in terms of IGS size, RFLPs and sequence data. Phylogenetic analyses (using CLUSTAL W, PHYLIP and TREEVIEW) indicated considerable differences between C. perniciosa strains from T. cacao and those from H. acutifolia (85-86%) and S. lycocarpum (95-96%). Sequence differences also indicated that C. perniciosa from T. cacao in Bahia is less variable (98%) when compared to the pathogen on T. cacao in Amazonas (97-98%), perhaps reflecting a recent introduction to T. cacao in Bahia. PMID- 12735242 TI - Cloning of and genetic variation in protease VCP1 from the nematophagous fungus Pochonia chlamydosporia. AB - The fungus Pochonia chlamydosporia is a biocontrol agent with commercial potential for root knot and cyst nematodes. It produces an alkaline serine protease, VCP1, during infection of nematode eggs. The gene encoding VCP1 was sequenced and the sequences of cDNAs from six isolates from different nematode hosts were compared. The gene encoding VCP1 was similar to PR1 from Metarhizium anisopliae with similar regulatory elements. Comparison of translated cDNA sequences revealed two amino acid polymorphisms at positions 65 and 99, indicating a difference between isolates from cyst and root nematodes. The positions and nature of the polymorphisms indicated that the two forms of VCP1 might have different properties and this was tested with five chromogenic polypeptide substrates. Enzyme assays revealed the two forms differed in their abilities to utilise Succ-Ala-Ala-Pro-Phe-pNa and Succ-Ala-Val-Pro-Phe-pNa, suggesting different amino acid affinities at the S3 binding region. This indicates host related genetic variation in VCP1 between isolates of P. chlamydosporia isolated from different nematode hosts, which might contribute to host preference. Such differences may be important in future exploitation of P. chlamydosporia as a nematode biocontrol agent. PMID- 12735244 TI - Quantitative inoculation of willow rust Melampsora larici-epitea with the mycoparasite Sphaerellopsis filum (teleomorph Eudarluca caricis). AB - The mycoparasite Sphaerellopsis filum (teleomorph Eudarluca caricis) was applied simultaneously with Melampsora larici-epitea on to willow leaf discs using eight concentrations of conidia. Inoculum densities were quantified and the numbers of uredinia of the rust, pycnidia and conidia of S. filum and rust spores produced per leaf disc were measured 13 d after inoculation (first assessment). Higher S. filum inoculum densities resulted in more rust uredinia being infected, but did not reduce the number of uredinia produced. The ratios of infected rust pustules: S. filum conidia applied were in a range of 0.25-0.31 when less than 20 S. filum spores were inoculated on to a leaf disc (0.95 cm2). Suppressive effects of S. filum on rust spore production were more obvious in the second assessment, carried out 23 d after inoculation. Inoculum densities of S. filum were significantly (P < 0.001) correlated with the frequency of uredinia infected (% variance accounted for [VAF] = 85.8), the number of S. filum pycnidia (% VAF = 81.4), S. filum spores produced (% VAF = 72.3) and rust spore production (% VAF = 48.6). Rust spore production was significantly (P < 0.001) negatively correlated with the frequency of uredinia infected (% VAF = 51.1), the number of S. filum pycnidia (% VAF = 42.0) and the number of S. filum spores produced (% VAF = 40.6). The best correlation was found between the number of pycnidia and the number of S. filum spores produced (% VAF = 88.8). PMID- 12735243 TI - Interactions between nematophagous fungi and consequences for their potential as biological agents for the control of potato cyst nematodes. AB - The efficacies of three nematophagous fungi, Paecilomyces lilacinus, Plectosphaerella cucumerina and Pochonia chlamydosporia, for controlling potato cyst nematodes (PCN) as part of an Integrated Pest Management (IPM) regime were studied. The compatibility of the nematophagous fungi with commonly used chemical pesticides and their ability to compete with the soil fungi Rhizoctonia solani, Chaetomium globosum, Fusarium oxysporum, Penicillium bilaii and Trichoderma harzianum were tested in vitro. Paecilomyces lilacinus was the most successful competitor when the ability to grow and inhibit growth of an opposing colony at both 10 and 20 degrees C was considered. P. lilacinus also showed potential for control of the soil-borne fungal pathogen R. solani, releasing a diffusable substance in vitro which inhibited its growth and caused morphological abnormalities in its hyphae. Pochonia chlamydosporia was least susceptible to growth inhibition by other fungi at 20 degrees in vitro, but the isolate tested did not grow at 10 degrees. Plectosphaerella cucumerina was a poor saprophytic competitor. Radial growth of Paecilomyces lilacinus and Plectosphaerella cucumerina was slowed, but not prevented, when grown on potato dextrose agar incorporating the fungicides fenpiclonil and tolclofos-methyl, and was not inhibited by the addition of pencycuron or the nematicide oxamyl. Radial growth of Pochonia chlamydosporia was partially inhibited by all the chemical pesticides tested. The efficacy of Paecilomyces lilacinus as a control agent for R. solani was further investigated in situ. Treatment with P. lilacinus significantly reduced the symptoms of Rhizoctonia disease on potato stems in a pot trial. The effectiveness of P. lilacinus and P. cucumerina against PCN was also tested in situ. Three application methods were compared; incorporating the fungi into alginate pellets, Terra-Green inoculated with the fungi and applying conidia directly to the tubers. Both formulations containing P. lilacinus and formulation mixtures alone, particularly alginate pellets, significantly reduced multiplication of PCN in soil. We conclude that P. lilacinus showed the greatest potential for use in combination with selected fungicides and nematicides as part of an IPM programme for the control of PCN, but further work is required to confirm whether it is effective against PCN in soil. PMID- 12735245 TI - Effect of mycoparasitic fungi on the development of Sphaerotheca fusca in melon leaves. AB - We evaluated the possibilities of exploitation of several mycoparasitic fungi in the biocontrol of cucurbit powdery mildew (Sphaerotheca fusca) in melon greenhouses. To simplify the screening, an in vitro biological control test on detached leaves of melon has been used and a detailed microscopic analysis of the interactions between mycoparasites and S. fusca conducted. In this context, the effect of mycoparasitic fungi on the formation of infection structures of S. fusca such as haustoria, conidia and conidiospores has been quantified. On the basis of the microscopic data, effect of mycoparasites on severity and incidence of cucurbit powdery mildew has been discussed. Our results show that, under controlled environmental conditions, Acremonium alternatum, Ampelomyces quisqualis and especially Lecanicillium lecanii, when applied in early stages of infection, were able to significantly reduce cucurbit powdery mildew symptoms and S. fusca development on melon leaves. These results indicate that these mycoparasites are promising candidates for the biocontrol of cucurbit powdery mildew in melon greenhouses. PMID- 12735246 TI - A high concentration of glucose inhibits Tuber borchii mycelium growth: a biochemical investigation. AB - Tuber borchii mycelium (strain 1BO) is able to utilise glucose, fructose or mannitol in the culture medium as a carbohydrate source. Since sugars not only function as a metabolic resource and structural constituent of cells, but also act as important regulators of various processes, we investigated if high sugar concentrations could influence fungal growth and development. The studies performed in this paper revealed that fructose or mannitol used at high concentration (50 g l-1) in the culture medium do not influence the growth and the biochemical responses of fungus but the growth of T. borchii mycelium is subject to glucose repression. In experiments with a high glucose concentration (50 g l-1) and with 2-deoxyglucose, a non-metabolisable glucose analogue, the growth of T. borchii was halved with respect to the control (10 g l-1 of glucose). The morphological and biochemical analyses revealed that the hyphae were metabolically and functionally active, but the activity of mannitol dehydrogenase was reduced to one-third in the high glucose treatment. This is the first evidence of glucose repression of growth and activity in the ascomycetous ectomycorrhizal fungus T. borchii. PMID- 12735247 TI - A functional interpretation of the role of the reticuloperidium in whole-ascoma dispersal by arthropods. AB - Auxarthron conjugatum (Onygenaceae) and Myxotrichum deflexum (Myxotrichaceae) are distantly related cleistothecial (gymnothecial) ascomycetes that form ascomata with strikingly similar peridia in which rigid, branched and anastomosed, thick walled hyphae create a cage- or mesh-like enclosure (reticuloperidium). We tested the hypothesis that the reticuloperidium plays a role in dispersal mediated by arthropods by enclosing ascomata of these fungi together with flies from the family Sarcophagidae. Gymnothecia of both fungi were picked up easily when the stiff hairs of the flies impaled the ascomata by passing through the interhyphal spaces of the reticuloperidium. Ascospore release from the gymnothecia then occurred during grooming activities during which the limbs of the flies caught the ascoma appendages causing the peridium to be torn apart. This adaptation to arthropod morphology and behaviour is interpreted as the driving force behind the evolution of reticuloperidia in unrelated groups of cleistothecial ascomycetes. PMID- 12735249 TI - Perenniporiella gen. nov. segregated from Perenniporia, including a key to neotropical Perenniporia species with pileate basidiomes. AB - Perenniporiella gen. nov. is segregated from Perenniporia. The new combinations Perenniporiella neofulva and Perenniporiella micropora, and the new species Perenniporiella pendula are proposed. Perenniporia piperis and Perenniporia albida are considered as taxonomic synonyms of P. neofulva. The three species are described and their taxonomic position is discussed. A key to the neotropical Perenniporiella and Perenniporia species with pileate basidiomes is presented. PMID- 12735248 TI - Viegasella and Mintera, two new genera of Parmulariaceae (Ascomycota), with notes on the species referred to Schneepia. AB - Two species previously accommodated in the genus Schneepia, S. pulchella and S. reticulata, are shown to belong to distinct new genera, described here as Viegasella and Mintera respectively. The new taxa are compared with Parmularia (of which Schneepia is confirmed as a synonym) and Symphaeophyma. Features of internal and external stromata, haustoria and stromata are emphasized as important characters for generic delimitation in the Parmulariaceae. The disposition of the remaining species assigned to Schneepia is discussed. PMID- 12735250 TI - Additional species of Cheiromycina (lichenized hyphomycetes), with a key to the known species. AB - Cheiromycina globosa and C. ananas spp. nov. are described in the lichenized hyphomycete genus Cheiromycina, one from Germany and one from the USA. A key is provided to all four species now known in genus, and all other lichenized hyphomycetes are briefly mentioned. All are corticolous and rarely recorded, so far only from temperate regions in Europe and North America. PMID- 12735251 TI - Native Venturia inopina sp. nov., specific to Populus trichocarpa and its hybrids. AB - Venturia populina, first described on European Populus nigra, has been thought to be the only species of Venturia in Europe and North America to cause leaf and shoot blight of balsam poplars and cottonwoods in Populus sects. Tacamahaca and Aigeiros. The species of Venturia occurring on introduced P. nigra and native P. trichocarpa in the Pacific northwest were examined. Venturia populina was consistently found on P. nigra (i.e. the widespread P. nigra cv. 'italica') in the region, but V. inopina sp. nov. was present on native P. trichocarpa and its hybrids. There were neither examples of V. populina on P. trichocarpa and its hybrids nor of V. inopina on P. nigra cv. 'italica' (27 collections from 16 sites in Oregon, Washington, and Vancouver Island were made during 1995-2002). In an inoculation study, host-range separation was confirmed in that V. inopina caused sporulating leaf lesions on P. trichocarpa and its hybrids, but only non sporulating lesions on P. nigra cv. 'italica'. These two species of Venturia can readily be distinguished by conidial septation; V. populina is primarily 2 septate, whereas V. inopina is primarily 1-septate. Growth rates on PDA at 15 degrees C, and ITS sequences (2.3% divergence) were also distinct in isolates of these congeners. Conidial shape was of more value in discriminant analysis than conidial length. Venturia inopina is homothallic, given the sexual fertility of cultures of single ascospores that were overwintered under ambient conditions. Its geographic range appears to be restricted even within the Pacific northwest, leaving open the possibility that still other undescribed, native species of Venturia occur elsewhere in North America on sects. Tacamahaca and Aigeiros. PMID- 12735252 TI - Coelomycetes in Dominican and Mexican amber. AB - Three species of fossil epiphyllous coelomycetes are described from Dominican and Mexican amber in the new genera, Asteromites gen. nov., Leptostromites gen. nov. and Leptothyrites gen. nov. Characters of the pycnidia and spores most closely resemble members of their extant respective genera, Leptostroma, Asteromella, and Leptothyrium, respectively. A. mexicanus sp. nov. occurs on a petal (possibly from a caesalpinoid legume such as Peltogyne) in Mexican amber. Leptostromites ellipticus sp. nov. occurs on a dicot leaf in Dominican amber, and Leptothyrites dominicanus sp. nov. on a monocot (grass?) leaf in Dominican amber. PMID- 12735253 TI - The oldest fossil myxogastroid slime mould. AB - A piece of Baltic amber (Tertiary, Eocene) contains a sporocarp of a slime mould which is assigned to the recent genus Arcyria and described as A. sulcata sp. nov. Apart from a fossil stemonitoid myxomycete, there are no further unambiguous fossil records of slime moulds and therefore the fossil gives new insights into the evolutionary history of the Myxomycetes. PMID- 12735255 TI - Prostate cancer testing. PMID- 12735256 TI - Complementary therapies in general practice. PMID- 12735257 TI - A GP's duty. PMID- 12735258 TI - Fits, faints and funny turns. A general diagnostic approach. AB - BACKGROUND: The patient presenting with a fit, faint or 'funny turn' can present a diagnostic dilemma for the general practitioner. OBJECTIVE: This article aims to provide an overview of the diagnostic approach to these 'episodes'. DISCUSSION: The key to diagnosis is to elicit a clear history focussing on the lead-up to the episode, a description of what took place and the events that took place after the episode. The patient's feelings, symptoms, circumstances and provocative factors give vital information. PMID- 12735259 TI - Dizziness and loss of consciousness. Cardiovascular causes. AB - BACKGROUND: Dizziness and loss of consciousness are common clinical problems presenting in general practice. OBJECTIVE: This article aims to provide the practitioner with a pragmatic and logical approach to identifying the cardiovascular causes of dizziness and loss of consciousness. DISCUSSION: A range of disorders with varying pathology cause a transient loss of consciousness associated with postural collapse (syncope) by interruption of blood flow to the brain. Syncope and seizures are the only common causes of recurrent episodes of loss of consciousness. The vasovagal reaction or 'common faint' and postural hypotension are both common and benign causes of syncope. Syncope can also result from cardiac causes that include disorders of cardiac rhythm and mechanical obstruction to cardiac output. Cardiac causes of syncope are associated with much higher morbidity and mortality than postural hypotension or fainting. Specific treatment is available for the various cardiac causes of syncope and thus accurate diagnosis is imperative. PMID- 12735260 TI - Fits, faints and funny turns. Could it be a mental disorder? AB - BACKGROUND: Patients who present to primary care with symptoms of fainting and dizziness, for which there is no adequate physical explanation, are frequently suffering from an undiagnosed psychiatric disorder. OBJECTIVE: This article aims to improve the recognition of common mental disorders presenting as 'fits, faints and funny turns' (FFFTs) and to encourage general practitioners to view these disorders as a positive diagnosis in need of treatment. DISCUSSION: Psychiatric disorders, particularly panic attacks and depression, are common in the setting of FFFTs and should be a positive diagnosis rather than a diagnosis of exclusion. A detailed description of the episode, with corroborating information from a witness if possible, and psychiatric rating scales can assist in this process. Identifying a physical cause for the episode does not exclude a psychiatric diagnosis and vice versa. Specific psychological and pharmacological therapies are effective for anxiety and depression. PMID- 12735261 TI - The new patient with a first seizure. AB - BACKGROUND: First seizures are common, with one in 20 people suffering a seizure at some time in their life. OBJECTIVE: This article aims to outline the assessment of patients with a first seizure, including making an accurate diagnosis of both seizure type and an epilepsy syndrome, if present. DISCUSSION: Seizures are classified into generalised and partial (arising from a focal region in the brain) based on clinical and electroencephalogram findings. However, as a partial seizure may proceed to a tonic clonic phase, differentiation may be difficult. Inquiring directly about 'minor' epileptic symptoms before the episode such as absences, myoclonic jerks, visual or auditory hallucinations or feelings of deja vu, is needed to attempt to make a epilepsy syndrome diagnosis, as this has practical implications for treatment, prognosis and genetic counselling. Generalised epilepsies should be treated initially with valproate, while partial epilepsies should be treated with carbamazepine and switched to newer agents if intolerance occurs. PMID- 12735262 TI - Use of antidepressant medications in the general practice setting. A critical review. AB - BACKGROUND: Antidepressants are commonly prescribed in general practice for depression, but also for a wide range of other psychiatric conditions and physical problems. OBJECTIVE: This review, although concentrating on the use of antidepressants in depression, also reviews their use in other conditions commonly seen in general practice. DISCUSSION: For most major depression, all antidepressant drugs have equal efficacy. The choice of antidepressant drug needs to be tailored to the particular patient's medical condition and personal preferences. It is likely that adverse effects are the major determinant in the choice of antidepressant for a particular patient. However, in treating conditions other than depression, the efficacy of the antidepressant drug can be the primary issue of drug choice. PMID- 12735263 TI - Therapeutic approach: how to get there. AB - BACKGROUND: After determining the therapeutic goals for an individual patient, the next step in good prescribing is planning a therapeutic approach to achieve these goals. OBJECTIVE: This article aims to illustrate the process of the therapeutic approach. An example of a patient with asthma is discussed, and the evidence base for the possible lifestyle interventions is reviewed. DISCUSSION: Clinicians often treat the therapeutic approach as a two step process of identifying nonpharmacological then drug related interventions. However, the scope of therapeutic approaches is really much broader than this. It encompasses lifestyle change, education, avoidance of triggers, procedural interventions, patient self management, as well as drug treatment. The latter may actually include medication cessation. PMID- 12735264 TI - Not just another fall in the elderly. Bilateral adrenal lymphoma presenting with adrenal insufficiency causing weakness. PMID- 12735265 TI - Sickness certificates. To write or not to write. AB - Sickness certificates are legal documents. General practitioners may face disciplinary proceedings, or even civil or criminal action if they issue a false, misleading or incorrect sickness certificate. Never backdate a sickness certificate. Always record the actual date that the certificate was written and the date of the consultation--even if the certificate refers to a different period of absence from work. PMID- 12735266 TI - Depression in adolescents. Key issues in assessment and management. AB - BACKGROUND: Mental disorders affect young people disproportionately and are increasing in prevalence. Knowledge about depression in adolescents and skill in its management lags behind that for adults. Depression is the most common mental disorder and untreated depression is associated with significant adverse effects. Depression is also a major predictor of suicidal behaviour and suicide. The role of the general practitioner in the treatment of adolescent depression is of considerable public health and clinical importance. OBJECTIVE: The aim of this paper is to highlight the major differences between adolescent and adult depression in relation to detection, assessment and treatment. DISCUSSION: The new Commonwealth Government's Better Outcomes in Mental Health Initiatives and Incentives provide an opportunity for GPs to improve their knowledge and skills in the detection, assessment and treatment of mental disorders. It is hoped that this paper will encourage GPs to include the topic of adolescent depression in their learning objectives. PMID- 12735267 TI - Dry eyes. Eye series. 3. AB - During her regular check-up a 68 year old woman patient mentions, as an aside, that the recent hot weather has left her eyes with a 'dry, gritty feeling'. The feeling is more noticeable toward the end of the day and on further questioning has gradually become more prominent over the past few years. PMID- 12735268 TI - Complex partial seizures. AB - A 24 year old woman with a past history of febrile convulsions presents with a history of several episodes of 'funny turns' over a period of six months. She states she 'feels weird' before each episode. Her partner describes that she becomes verbally unresponsive, licks her lips, and makes other strange movements with her mouth. They usually last several minutes, after which she appears quite drowsy for half an hour. She recalls nothing of the event itself. Examination findings are normal. You take blood tests and arrange for an electroencephalogram. PMID- 12735269 TI - Sustaining quality general practice in a flawed market. AB - BACKGROUND: Recent years have seen an increasing corporatization of general practice. OBJECTIVE: This paper considers how divisions of general practice should respond to the corporatization of general practice. DISCUSSION: Corporate models are an economically rational outcome of structural features of Medicare funding for Australian general practice. One limitation of these structures is that they are indifferent to quality. Corporate general practice competes with divisions of general practice at some level because it provides an alternative organisational structure and service values for large numbers of general practitioners. Accepting this, the challenge for divisions becomes one of differentiating themselves from the corporate model through a focus on promoting and encouraging quality practice. PMID- 12735270 TI - High blood pressure research in Australian general practice. PMID- 12735271 TI - Patients' views of high blood pressure, its treatment and risks. AB - OBJECTIVE: To examine the understanding and beliefs of a sample of Australian patients about high blood pressure and its therapy, and to examine the accuracy of their assessment of their own risk. METHOD: A cross sectional study in two general practices in Perth, Western Australia. Interviews were conducted with 55 consecutive patients aged 40-80 years of age with uncomplicated hypertension. Qualitative and quantitative data were collected on patients' beliefs about the nature, symptoms, causes and treatment of high blood pressure as well as their adherence to antihypertensive medications during the preceding month. Patients also estimated their own risk of stroke or myocardial infarction. RESULTS: Two thirds of the patients (65%) described high blood pressure within an appropriate biomedical definition. Forty-five percent attributed a variety of symptoms to their high blood pressure while 55% believed that stress was a cause of their high blood pressure. Three-quarters (73%) were fully adherent to their medications in the preceding month. Seventy-one percent and 62% of patients were aware that stroke and heart attack respectively are possible consequences of high blood pressure. They significantly overestimated their risk of stroke and myocardial infarction. CONCLUSION: One-third of treated hypertensive patients are poorly informed about the causes and effects of hypertension, and overestimate their risk of both stroke and heart attack, as well as the benefit derived from treatment. PMID- 12735272 TI - The management of dementia in general practice. A field test of guidelines. AB - INTRODUCTION: Guidelines for the management of dementia in non-institutionalized patients living in the community were developed by a broadly representative group. We assessed their usefulness. METHOD: The draft guidelines included emphasis on psychosocial issues. They were field tested by 17 general practitioners with 119 dementia patients. RESULTS: There was a high prevalence of comorbidity in the patients and frequent psychosocial issues in their management that were often not addressed. The guidelines were rated as very helpful for at least one aspect of care for 50% of the patients. DISCUSSION: The guidelines were found to be useful to GPs. PMID- 12735273 TI - Trends and geographic variations in hospital admissions for asthma in Victoria. Opportunities for targeted interventions. AB - OBJECTIVE: To describe variations in rates of hospital admissions for asthma in Victoria as health indicators of quality of primary care services and access. DESIGN: Routine analyses of age and sex standardised admission rates of asthma in rural and metropolitan Victoria from 1993-1994 to 1999-2000. RESULTS: There were 10,079 admissions for asthma, with an average of 2.71 bed days in 1999-2000. The admission rate for asthma decreased from 3.1/1000 (95% CI: 3.1-3.2) in 1993-1994 to 2.2/1000 (2.1-2.2) in 1999-2000, with a 37% reduction in rural regions and 26% in metropolitan regions. Sixteen primary care partnerships (small areas), 13 of them rural, had significantly higher admission rates than the Victorian average. CONCLUSION: Although asthma hospital admission rates are falling faster in rural than metropolitan areas, rural areas still have higher admission rates with significant variation between small areas. PMID- 12735274 TI - Instrumental and Pavlovian incentive processes have dissociable effects on components of a heterogeneous instrumental chain. AB - Two experiments compared the effects of Pavlovian stimuli and incentive learning on the performance of a heterogeneous chain of instrumental actions. Using a Pavlovian-instrumental transfer design, the authors found that only a stimulus paired with the same outcome as that earned by performance of the chain produced positive transfer, an effect that was restricted to the action in the chain most proximal to reward delivery. In contrast, after a shift to a nondeprived state, only animals that had previously consumed the instrumental outcome when they were nondeprived decreased instrumental performance. Furthermore, this effect of the incentive learning treatment was limited to performance of the distal action. Together these data suggest that Pavlovian and instrumental incentive manipulations have dissociable effects on instrumental performance. PMID- 12735275 TI - Discrimination of structure: II. Feature binding. AB - In 3 experiments, pigeons acquired a discrimination between patterns comprising the same features. Thus vertical green bars beside horizontal red bars might have signaled food, and horizontal green bars beside vertical red bars might have signaled no food. The solution of this discrimination can be explained by assuming each pattern is represented either by a template or by structural features that are sensitive to combinations of color and line orientation. The 1st explanation predicts subjects should react to a training pattern rotated 90 degrees in the same way as the pattern on which it is based. The 2nd explanation predicts these patterns should be treated as if they signal opposing outcomes. The experiments confirmed the 2nd of these predictions. PMID- 12735276 TI - Visual search asymmetry in pigeons. AB - Pigeons received an odd-item search task that involved an array of 12 patterns containing 11 similar distractors and a single target. Pecks to the target resulted in the delivery of food. Accuracy was greater on trials when a distinctive feature was located in the target but not in the distractors, rather than when the feature was in the distractors but not in the target. This search asymmetry was influenced by the similarity of the target to the distractors. The results are similar to those obtained with humans. PMID- 12735277 TI - Temporally graded, context-specific retrograde amnesia and its alleviation by context preexposure: effects of postconditioning exposures to morphine in the rat. AB - Five experiments studied retrograde impairments in Pavlovian fear conditioning following prolonged exposure to the opioid receptor agonist morphine. Injections of morphine commencing 1-7 days but not 14 days after conditioning produced amnesia for that conditioning episode. This amnesia was (a) selective such that morphine impaired freezing to the conditioning context but not to the auditory conditioned stimulus, (b) independent of the interval between the last injection of morphine and test, and (c) accompanied by a failure of contextual discrimination. Context preexposure protected context conditioning and discrimination from the amnestic effects of morphine. These results show that retrograde deficits in contextual fear conditioning are mediated by failures to consolidate a contextual representation. PMID- 12735278 TI - Intradimensional and extradimensional shifts in spatial learning. AB - Animals trained on 2 discriminations learn the 2nd rapidly if the relevant stimuli are from the same dimension as the 1st (an intradimensional or ID shift) but slowly if the relevant stimuli for the 2 problems are from different dimensions (an extradimensional or ED shift). Four experiments examined ID and ED shifts in spatial learning. Rats trained on 2 spatial problems learned the 2nd more rapidly than rats whose 1st problem had been nonspatial. But this difference between ID and ED shifts depended on the spatial relationship between rewarded (S+) and unrewarded (S-) alternatives in the 2 spatial problems. The results imply that rats trained on a spatial discrimination do not learn to attend to all spatial landmarks but only to those that serve to differentiate S+ and S-. PMID- 12735279 TI - Facilitation of extinction by an increase or a decrease in trial duration. AB - Five experiments examined the effects of altering the duration of a conditioned stimulus (CS) for extinction. For the first 3 experiments, rats received conditioning with a 10-s CS before different groups received extinction with a CS that was either the same duration or longer than that used for conditioning. For the remaining 2 experiments, conditioning was conducted with a 60-s CS before different groups received extinction with a CS of either the same duration or a shorter duration than that used for conditioning. In all experiments, extinction progressed more readily when the CS duration was different for the 2 stages than when it was constant. The results are discussed in terms of rate expectancy theory and associative learning theory. PMID- 12735280 TI - [Pursuing the future]. PMID- 12735281 TI - [Computerized monitoring for integrated cervical screening. Rationale, methods and indicators of participation]. AB - This paper presents a computerised system for the monitoring of integrated cervical screening, i.e. the integration of spontaneous Pap smear practice into organised screening. The general characteristics of the system are described, including background and rationale (integrated cervical screening in European countries, impact of integration on monitoring, decentralised organization of screening and levels of monitoring), general methods (definitions, sections, software description, and setting of application), and indicators of participation (distribution by time interval since previous Pap smear, distribution by screening sector--organised screening centres vs public and private clinical settings--, distribution by time interval between the last two Pap smears, and movement of women between the two screening sectors). Also, the paper reports the results of the application of these indicators in the general database of the Pathology Department of Imola Health District in northern Italy. PMID- 12735282 TI - [Expression of p27 and proliferative (MIB-1), mitotic (MI) and apoptotic indices in early-phase (EGF) gastric carcinoma. Results of a study by the Italian Gastric Cancer Research Group (IRGGC)]. AB - AIMS: Since the Japanese Society for Gastroenterology and Endoscopy (JSGE) introduced the definition of Early Gastric Cancer (EGC), much more and deeper studies were done, which demonstrated that EGC was a more complex phase of the neoplastic disease with different morphologic characteristics, tightly linked to the prognosis. We evaluated the clinical impact of some prognostic factors, known being important in the advanced lesions, in a series of EGC patients with special reference to the clinicomorphological features. METHODS AND RESULTS: We analysed the mitotic (MI) and apoptotic (AI) indices and the immunohistochemical expression of p27 and MIB-1 in 83 EGC cases consecutively recruited in the hospitals of Forli, Verona, Siena and Milan (IRGGC) in the period 1994-95. The classifications of JSGE, Lauren and Kodama were used to define the macroscopic, microscopic and growth pattern types, respectively. Decreased p27 expression correlated with the macroscopic escavated lesions and diffused mixed histotypes; the increase of MIB-1 detection with tumour size larger than 2 cm, but lesser than 4 cm; MI with intestinal histologic types and AI with mucosal and penetrating lesions, according to Kodama. Statistical analysis showed significative correlations among MIB-1, MI and AI, but not with p27 and the other variables. All these factors did not influence the prognosis of our patients. CONCLUSIONS: In our series, p27, MIB-1, MI, and AI did not add any useful clinical. So, in EGC patients the morphological features have still the most important role in influencing the prognosis and treatment of patients. PMID- 12735284 TI - [Thin-layer cytology in urinary diagnosis]. AB - To evaluate the thin-layer method in urinary cytology we have compared cytospin smears, filter and thin layer. Cellularity, cellular morphology and background were evaluated. Thin layer method in urinary cytology improve cell recovery and cell preservation, reducing background artefact. Cytohistologic correlations with thin layer method were better than with filter and cytospin; an higher capacity to detect low grade transitional cell carcinomas, frequently undiscovered by cytology, was demonstrated. PMID- 12735283 TI - [Expression of steroid receptors and DNA synthesis in male breast cancer]. AB - Using a double-labeling immunocytochemical-autoradiographic assay we studied 18 male breast carcinomas to evaluate the cell kinetic and receptor status in neoplastic cells during S-Phase and to detect possible differences with respect to 21 female breast cancers, from a previously, published study. In male breast cancer, the tumor receptor content and ER/PgR expression in neoplastic cells during S-Phase was higher (p = 0.01) than that in corresponding female tumor while tumoral cell proliferation was lower, but not significantly. In the previous reported study on female breast cancer we demonstrated that proliferative activity was higher in receptor negative cell population both for ER and PgR. Conversely, in male tumor, that difference was only present in relation to the expression of PgR: in fact, the proliferative activity was higher in PgR negative than in PgR positive cells (Anova Test: p = 0.04) while no difference was evidenced between ER negative versus ER positive cells. Moreover, the arrest of DNA synthesis, expressed as percentage of cells without 3H-Tdr labeling, was not related to either the ER or PgR expression, while in female breast cancer it was higher in PgR positive than ER positive cell population. Our data confirmed differences between males and females regarding the receptor status and cell cycle S-Phase in breast cancer. The poorer prognosis of the male breast carcinoma might be related to ineffective therapies which do not consider these differences in the biological profile of the male tumor. The Authors indicate that prognostic and predictive tissutal markers, detected by immunocytochemical methods and useful for therapeutic programming in the female breast cancer have a different significance in the male breast cancer and stress the need for different therapeutic strategies specific for male breast cancer. PMID- 12735285 TI - [Ascertainment of death: a certain death and ascertained death]. PMID- 12735286 TI - [Round atelectasis of the lung: clinicopathological study of 6 cases and review of the literature]. AB - OBJECTIVE: To report about 6 new patients with round atelectasis of the lung, 1 of them professionally exposed to asbestos and another to silicates. RESULTS: The patients, 5 males and 1 female, presented with a peripheral, rounded pulmonary opacity, simulating a neoplasm. The examination of the surgical specimen revealed a pleuritis, with multiple pleural folding: the underlying lung parenchima was compressed, but otherwise unremarkable. CONCLUSIONS: Round atelectasis is relatively unusual for the pathologist. However, the correct diagnosis is potentially important, because the lesion can be the sign of a significant asbestos exposure. PMID- 12735287 TI - [Hepatic splenosis diagnosed by fine-needle cytology]. AB - A case of hepatic splenosis, diagnosed by FNA cytology, in a 56 years old cirrhotic man is reported. The cytologic material, aspirated from a 4.8 cm "suspicious for hepatocarcinoma" nodule, showed, within a hematic background, small-medium sized lymphoid cells, neutrophils and eosinophils granulocytes, histiocytes with large cytoplasm, plasma cells; mixtures of these cells, within a reticular network, were often aggregated in large structures, in which endothelial cells organized in vascular-like structures were also visible. The cell block, showed sinusoidal structures, lymphoid follicular aggregates and fibrous septa, strongly suggesting the micromorphology of the normal spleen. Splenosis is an autotransplant of splenic tissue, morphologically organized in non-capsulated, functioning, self-limited nodules, secondary to a post-traumatic splenectomy. Usual presentation of this condition is the presence of several bluish nodules on the peritoneal surface. Nevertheless, sporadic cases in other organs have been reported. To the best of our review of the Medical Literature, a hepatic splenosis is an exceptional even and the present is the first case in this location diagnosed by FNA cytology. The present one is also the case with the largest latency between the splenectomy and the splenosis detection (43 years). PMID- 12735289 TI - [Use of denatured ethyl alcohol in the pathological anatomy laboratory]. PMID- 12735288 TI - [Primary adrenal angiosarcoma]. AB - Primary angiosarcomas of the adrenal gland are extremely rare neoplasms, first described in 1988. We report the twentieth case, incidentally discovered in a 60 year-old male. The main histological features were epithelioid appearance and immunohistochemical co-expression of endothelial markers and cytokeratin by neoplastic cells. The potential diagnostic pitfalls of these morphological characteristics are discussed. The patient underwent surgery and adjuvant chemotherapy and is free of tumour recurrence three years after surgery. PMID- 12735290 TI - [From asklepieia to monasteries: the places of medical art in Antiquity]. AB - The article deals with the places of medical art in the antiquity. Author's intention is to give a description of the places where developed the assistance and welfare activity to sick people from the Asklepieia to Monastic Hospitals. In Theurgical Medicine the cure for sickness was peculiar to gods, at first to all gods, mostly to Apollo and Artemide; later healing art had an own god: Asclepius. In the temples of Asclepius, the Asklepieia, prayers, sacrifices, offerings and magical rituals began to be associated with medical practical exercise and rational therapeutic systems. Rational Hippocratic Medicine found own places in the cities: the iatreia in Greece and the tabernae medicae in Rome. The article tries to describe the evolution of social welfare assistance from the Roman Valitudinaria to the monastic xenodochia and the first forms of Religious Hospitals. PMID- 12735292 TI - Fabry disease in genetic counseling practice: recommendations of the National Society of Genetic Counselors. AB - The objective of this document is to provide health care professionals with recommendations for genetic counseling and testing of individuals with a suspected or confirmed diagnosis of Fabry disease, with a family history of Fabry disease, and those identified as female carriers of Fabry disease. These recommendations are the opinions of a multicenter working group of genetic counselors, medical geneticists, and other health professionals with expertise in Fabry disease counseling, as well as an individual with Fabry disease who is a founder of a Fabry disease patient advocacy group in the United States. The recommendations are U.S. Preventive Task Force Class III, and they are based on clinical experience, a review of pertinent English-language articles, and reports of expert committees. This document reviews the genetics of Fabry disease, the indications for genetic testing and interpretation of results, psychosocial considerations, and references for professional and patient resources. These recommendations should not be construed as dictating an exclusive course of management, nor does use of such recommendations guarantee a particular outcome. The professional judgment of a healthcare provider, familiar with the facts and circumstances of a specific case, will always supersede these recommendations. PMID- 12735293 TI - Biochemical characterization of proline racemases from the human protozoan parasite Trypanosoma cruzi and definition of putative protein signatures. AB - Proline racemase catalyzes the interconversion of L- and D-proline enantiomers and has to date been described in only two species. Originally found in the bacterium Clostridium sticklandii, it contains cysteine residues in the active site and does not require co-factors or other known coenzymes. We recently described the first eukaryotic amino acid (proline) racemase, after isolation and cloning of a gene from the pathogenic human parasite Trypanosoma cruzi. Although this enzyme is intracellularly located in replicative non-infective forms of T. cruzi, membrane-bound and secreted forms of the enzyme are present upon differentiation of the parasite into non-dividing infective forms. The secreted form of proline racemase is a potent host B-cell mitogen supporting parasite evasion of specific immune responses. Here we describe that the TcPRAC genes in T. cruzi encode functional intracellular or secreted versions of the enzyme exhibiting distinct kinetic properties that may be relevant for their relative catalytic efficiency. Although the Km of the enzyme isoforms were of a similar order of magnitude (29-75 mM), Vmax varied between 2 x 10(-4 )and 5.3 x 10(-5) mol of L-proline/s/0.125 microM of homodimeric recombinant protein. Studies with the enzyme-specific inhibitor and abrogation of enzymatic activity by site directed mutagenesis of the active site Cys330 residue reinforced the potential of proline racemase as a critical target for drug development against Chagas' disease. Finally, we propose a protein signature for proline racemases and suggest that the enzyme is present in several other pathogenic and non-pathogenic bacterial genomes of medical and agricultural interest, yet absent in mammalian host, suggesting that inhibition of proline racemases may have therapeutic potential. PMID- 12735294 TI - A survey of genetic counselors' strategies for addressing ethical and professional challenges in practice. AB - There is limited research about ethical and professional dilemmas that genetic counselors encounter in their practice and their strategies for addressing them. In this study, 454 genetic counselors rated the frequency with which they encounter each of 16 ethical/professional challenges identified and categorized previously (McCarthy Veach P., Bartels DM, LeRoy BS (2001) J Genet Couns 10(2):97 119). Over 40% indicated these issues occurred frequently: patient emotions, diversity, financial constraints, uncertainty, and colleague error. Two hundred and fifty-five respondents provided personal anecdotes describing exceptionally challenging situations and recommended strategies for addressing them. Most of their anecdotes involved informed consent, value conflicts, confidentiality, colleague error, withholding information, and resource allocation. The most frequently recommended strategies were further discussion with patients, consultation with other professionals, and referral to other health sources. Thirty-five respondents were unable to/did not offer strategies. Respondent demographics were not related to frequency of issues, type of anecdote, or recommended strategies. Practice, policy, and research implications are discussed. PMID- 12735295 TI - Models of genetic counseling and their effects on multicultural genetic counseling. AB - This theoretical paper examines challenges to multicultural genetic counseling, counseling between culturally different clients and counselors, in the context of Kessler's typology of models of genetic counseling (Kessler S (1997) J Genet Counsel 6:287-295). It is suggested that challenges such as resistance to multicultural genetic counseling education may be due to conceptions about genetic counseling as a biomedical field that transcends questions of culture as well as lack of multicultural training or prejudice. Directions for future research and recommendations for multicultural genetic counseling education are briefly explored. PMID- 12735296 TI - DNA-sequence patenting: National Society of Genetic Counselors (NSGC) position paper. AB - In November 2000, the Genetic Services Committee of the National Society of Genetic Counselors (NSGC) convened a working group to draft a position paper on patenting DNA-sequences. The mandate of the group was to produce general position statements that support the perspective and needs of consumers of DNA-based genetic tests and therapies (our patients and their families) and participants in DNA-based genetic research. After review and discussion of the literature on DNA sequence patenting issues, the working group drafted position statement points that support current United States Patent and Trademark Office (USPTO) guidelines; broad licensing of DNA-sequence patents; nonenforcement of DNA sequence patents in noncommercial research; reasonable royalty rates; an informed consent process for research participants that discloses whether they can share in any financial rewards relating to the project; the development of guidelines for licensing of DNA-sequence patents; and the establishment of oversight organizations to monitor licensing of DNA-sequence patents. These position statements were approved by the NSGC Board of Directors in the fall of 2001. PMID- 12735297 TI - Some genetics for some Jews: a Jewish look at the human genome. PMID- 12735298 TI - Bioethics, the genome, and the Jewish body. PMID- 12735299 TI - Cross-generational retribution and genetic engineering: reflections on chance and free will. PMID- 12735300 TI - The human genome and ethical issues. PMID- 12735301 TI - [Meningococcal purpura fulminans]. PMID- 12735302 TI - The neural progenitor-restricted isoform of the MARK4 gene in 19q13.2 is upregulated in human gliomas and overexpressed in a subset of glioblastoma cell lines. AB - Alterations of 19q13 are frequently observed in glial neoplasms, suggesting that this region harbors at least one gene involved in gliomagenesis. Following our previous studies on structural 19q chromosome rearrangements in gliomas, we have undertaken a detailed FISH analysis of the breakpoints and identified a 19q13.2 intrachromosomal amplification of the MAP/microtubule affinity-regulating kinase 4 (MARK4) gene in three primary glioblastoma cell lines. Recent data suggest that this gene is involved in the Wnt-signaling pathway. We observed that the expression of the alternatively spliced MARK4L isoform is upregulated in both fresh and cultured gliomas and overexpressed in all of the above three glioblastoma cell lines. Interestingly, we also found that MARK4L expression is restricted to undifferentiated neural progenitor cells or proliferating glial precursor cells, whereas its expression is downregulated during glial differentiation. Perturbation of expression using antisense oligonucleotides against MARK4 in glioblastoma cell lines, consistently induced a decreased proliferation of tumor cells. Taken together, these data show that MARK4, which is normally expressed in neural progenitors, is re-expressed in gliomas and may become a key target of intrachromosomal amplification upon 19q rearrangements. PMID- 12735303 TI - Flavopiridol in untreated or relapsed mantle-cell lymphoma: results of a phase II study of the National Cancer Institute of Canada Clinical Trials Group. AB - PURPOSE: To determine the response rate and toxicity of flavopiridol in patients with previously untreated or relapsed mantle-cell lymphoma. PATIENTS AND METHODS: Adult patients with previously untreated or in first or second relapse of previously responsive mantle-cell lymphoma were given flavopiridol 50 mg/m2/d by intravenous bolus for 3 consecutive days every 21 days with antidiarrheal prophylaxis. Flavopiridol was continued until disease progression, unacceptable toxicity, or stable disease for four cycles. Disease was reassessed every two cycles. RESULTS: From 33 registered patients, 30 were eligible after pathology review, 30 were assessable for toxicity, and 28 were assessable for response. A median of four cycles of treatment was administered; 90% of patients received at least 90% of planned dose-intensity. No complete responses were seen; three patients had a partial response (11%), 20 patients had stable disease (71%), and five patients had progressive disease (18%). The median duration of response was 3.3 months (range, 2.8 to 13.2 months). The most common toxicities were diarrhea (97%), fatigue (73%), nausea (47%), and vomiting (27%). At least one nonhematologic grade 3 or 4 toxicity was seen in 14 patients (47%). Hematologic toxicity was modest. CONCLUSIONS: Flavopiridol given as a daily bolus for 3 consecutive days every 3 weeks has modest activity as a single agent for mantle cell lymphoma. The number of stable and partial responses that was seen indicates that it is biologically active and may delay progression. Future studies in mantle-cell lymphoma should test this agent with other active agents and using different schedules. PMID- 12735304 TI - Is the brain overrated? PMID- 12735305 TI - Outreach critical care-cash for no questions? PMID- 12735306 TI - Outreach critical care-cash for no questions? PMID- 12735307 TI - Neurological deterioration after laryngeal mask insertion. PMID- 12735308 TI - Six barriers to veterinary career success. PMID- 12735309 TI - [Classification of acquired colonic strictures (according to literature reviews and the author's own observations)]. PMID- 12735310 TI - [Physico-technological conditions of radiography at the national diagnostic radiological facilities "Medics-R" and "Telemedics-R"]. PMID- 12735311 TI - Histocompatibility antigens and humoral immunity to Epstein-Barr virus. AB - To explore possible genetic determinants of immunity to Epstein-Barr (EB) virus infection, prevalence rates and geometric mean titers (GMT) of antibody against EB viral capsid antigen (VCA) were measured in 422 adults of known HLA and ABO histocompatibility types. Deviation from the overall prevalence (87%) of anti-VCA titers > 10 was limited to subjects with HLA-A10 (95%) and those with a "blank" HLA-A locus (78%). The latter deviation was largely accounted for by the 13 subjects of HLA-A1/blank constitution (46%). Deviations from the overall mean anti-VCA titer (GMT 131.5) were found in seropositive subjects with blood group 0 (GMT 153.3) and tissue antigens HLA-A3 (GMT 150.8), HLA-B7 (GMT 152.1), HA-Bw15 (GMT 97.0), and HLA-B27 (GMT 86.4). Separate reanalysis of the data for male and female subjects verified the association of blood group 0 with elevated and HLA B27 with reduced mean levels of anti-VCA antibody. PMID- 12735312 TI - HLA-D region products are expressed in endothelial cells. AB - The mixed lymphocyte endothelial cell culture was studied by the primed lymphocyte typing (PLT) technique. By comparing the HLA-D/DR specificity of the secondary response when using either peripheral blood mononuclear cells (PBM) or endothelial cells from umbilical cords for priming or restimulation of lymphocytes, it was found that PBM from newborn would induce a clear-cut specificity for HLA-D/DR when used for priming a well a for restimulation. No HLA D/DR specificity was seen, however, when endothelial cells were used for restimulation of lymphocytes primed to HLA-D/DR on PBM. On the other hand, lymphocytes primed to endothelial cells showed significant, albeit not very strong specificity for HLA-D/DR when restimulated with PBM. Our experiments suggest that HLA-D region products are present on endotheial cells, and thus confirm and extend serological studies using anti DR antisera. PMID- 12735313 TI - Data reduction in HLA-D typing with the primed lymphocyte typing (PLT) technique. the normalized median response (NMR). AB - We describe here a simple non-parametric method for the evaluation of data obtained from primed lymphocyte typing (PLT) experiments. The method is designed to compensate for both differences in the PLT responsiveness and stimulatory capacity by a double normalization procedure based on a representative negative reaction (the median) for each PLT-cell and for each secondary stimulator. The index of response is called the Normalized Median Response (NMR). The NMR-method was developed from data from one of a series of PLT experiments aimed at HLA-D typing. The NMR-method was applied to eight other PLT experiments and was compared to two other calculation methods based on the maximal response of each PLT-cell: (i) a method without compensation for stimulator variation, and (ii) a method including non-parametric compensation for stimulator variation. The eight experiments involved 37 different PLT cells tested against 74 different stimulating cells yielding a total of 1,334 secondary combinations. The NMR method descriminated significantly better than the other methods when the results were compared to the HLA-D types obtained with homozygous typing cell technique. In particular, the number of extra positive reactions was diminished by the NMR method. The reproducibility and the sources of variation were studied in the eight experiments and in five additional experiments. The NMR-method reduced the combined priming-to-priming variation and day-to-day variation to a level comparable to the variation between duplicate testings of the same PLT cell tested towards the same set of secondary stimulators in the same secondary PLT experiment. PMID- 12735314 TI - Generation of HLA-D specific primed lymphocyte typing (PLT) cells and cross reactions of PLT-cells primed with homozygous typing cells. AB - An approach for the selection of HLA-D specific primed lymphocyte typing (PLT) cells is described. The responder cells were primed with homozygous typing cells. Reproducible extra reactions were found and were analyzed in relation to HLA-D antigens defined by homozygous in cells (HTC's). The secondary response of 105 different PLT-cell combinations generated by 29 different primary responders against 19 different homozygous typing cells of the specificifies Dw1 to Dw8 and the local specificity "H" were tested in secondary PLT toward 17 different homozygous typing cells and 10 heterozygous cells. Cross-reactions were defined as reactions equal to or higher than the lowest HLA-D specific reaction observed. The entire experimental design and data analysis gave rise to a conservative definition of cross-reactivity. Two main groups of cross-reacting HLA-D determinants seem to exist: (i) Dwl, 3, 4, 7, and the local specificity "H", and (ii) Dw2, 5, 6, 8, and "H". The primary pairwise cross reactions were in group (i): Dw1-3, Dw1-"H", Dw3-4, Dw3-7, Dw7-"H", and in group (ii): Dw2-6, Dw2-8, Dw5 8, and Dw5-"H". The existence of such cross-reactions is likely to interfere with the results of PLT-typing and should be taken into account when attempts are made to develop HLA-D specific PLT-cells. PMID- 12735316 TI - Functional study and detection of HLA-D products on fractionated human bone marrow cells. AB - Bone marrow cells from nine normal human volunteers obtained from the Iliac crest, were used in this work for antigen determination and functional studies. The bone marrow aspirated cells were sequentially separated: elimination of erythrocyte, granulocytes and monocytes achieved by Ficoll-Isopaque centrifugation followed by plastic adherence. Purified bone marrow cells were finally separated by size using velocity sedimentation. The slow sedimenting small cells were shown to be mainly T lymphocytes, probably of blood origin. The medium sized bone marrow cells were shown to contain myeloid precursors (CFu-c). Large immature cells were in cycle actively synthesizing DNA molecules. HLA-D and HLA-DR detections on the fractionated cells were performed using three techniques: fluorescence with specific anti HLA-DR allo and xeno antisera; primed lymphocyte typing (PLT) with anti HLA-DR monospecific in vitro primed lymphocytes and detection of the HLA-D stimulating product using the bone marrow fractionated cells as stimulators in a mixed leukocyte culture. Concordant results were obtained with the three techniques. Lymphocytes in the bone marrow express HLA-D products a peripheral lymphocytes. Bone marrow fractions depleted of lymphocytes and monocytes also contain approximately 20% of cells expressing HLA-D products The meaning of the expression of HLA-D products on immature precursors non lymphoid cells is discussed. PMID- 12735315 TI - Use of monocytes in HLA-A, B, C and DR typings. AB - A simple method of improved serologic typing of monocytes for HLA-A, B, C and DR specificities is described. The method employs monocytes recovered from frozen samples of peripheral blood mononuclear cells; it chiefly involves pretreatment of monocytes with 0.01% iodoacetamide (IAA) prior to typing. The advantage of this method lies principally in the lowering of the background nonspecific cytotoxicities and false positive readings upon IAA addition to the monocyte preparations. Using this method monocytes can be typed for HLA-A, B, C determinants. Although the addition of IAA results in substantial typing improvements, we found the assignment of A, B, C specificities difficult due to the presence of extra positive reactions when monocytes were compared to T lymphocyte typings. probably due to the presence of DR or monocyte specific antibodies in the routinely used HLA antisera. This method proved to be most useful in DR typings where mono cytes in the presence of IAA were compared with autologous B cells in the absence of IAA. The differences in typings due to a decrease in false positive cytotoxic readings were significantly in favor of using IAA treated monocytes in DR typings (P < 0.0001). The use of IAA in the course of B cell or T cell typings bad no adverse consequences on either A, B, C or DR typings, respectively. Our results indicate a potential usefulness for the use of IAA in typing monocytes HLA determinants in general and for the DR determinants in particular. PMID- 12735317 TI - HLA-DRw3 in juvenile onset diabetes mellitus in Chinese. AB - Thirty-nine juvenile onset diabetes mellitus (JOD) patients and 57 healthy Chinese were HLA-typed in Taiwan. The incidence of HLA-DRw3 was significantly increased in patients when compared with the control group (R.R. = 5.8, P = 0.0027). The frequencies of HLA-B17, which showed linkage disequilibrium with DRw3, and Bw54 were increased in patients, but these increases were not significant because of the small number tested. The DR antigen associated with JOD was the same among Chinese and Caucasians, which suggested that at least one of the susceptibility genes to JOD would be commonly in linkage disequilibrium with DRw3 in the two populations. PMID- 12735318 TI - Genetics of cell-mediated lympholysis (CML) in man. AB - Human cytotoxic lymphocytes (CTL), developed in responder-stimulator combinations incompatible for only one HLA-B antigen, have been used to study the specificity and genetics of target cell determinants. We were able to develop specific CTLs directed against HLA-B7, B8, B27 and Bw44, as defined by standard serological reagents. CTL-anti-B5 and CTL-anti-Bw35 gave a number of false positive reactions. In addition, we have found that specific cytotoxic effector cells can be generated against B27 by sensitization of cells from a B7 donor and vice versa; B7 and B27 can in this way be recognized easily. The data support the view that HLA-B is the target antigen. In addition, population and family studies revealed that extra reactions with CTLs developed in female (parous) responder male stimulator combinations were due to HLA-restricted H-Y antigen killing. These findings suggest that by selecting responder-stimulator pairs differing for only one antigen at a locus, it should be possible to develop CTLs which will allow recognition of other specificities at the HLA-B locus and other loci. PMID- 12735319 TI - Human B-blast specific target determinants in CML: a methodological study. AB - A method for preparation of purified human PWM stimulated B lymphoblasts and their usefulness as targets in CML is described. The purified B lymphoblasts were found to carry surface membrane immunoglobulin (SmIg) and HLA-DR antigens and were non E-rosette forming. In contrast, PHA-stimulated lymphoblasts were found to be E-rosetting, SmIg negative and without serologically detectable HLA-DR antigens, and thus were characterized as T lymphoblasts. Using B and T lymphoblasts in parallel as targets in CML, it was possible to demonstrate target determinants exclusively expressed by the B lymphoblasts. PMID- 12735320 TI - Human B-blast specific target determinants in CML: a panel study. AB - The B blast specific target determinant phenotype defined by cytotoxic lymphocytes and the HLA-DR phenotype was tested for correlation in two panel studies. First, CTL reagents with B blast specificity were selected from CTLs educated between HLA-A, B, (C) identical unrelated donors giving no cytotoxicity with specific T blasts, resulting in free selection of panel B blasts used as targets. Second, CTLs educated between HLA non-identical unrelated donors giving cytotoxicity against specific T as well a B blast targets could be seen with negative reactions against T blasts but positive reactions against B blasts of selected panel members giving, in this set up, a free selection of donors cocultured for education of CTLs. These panel studies showed that HLA-DR associated B blast specific target determinants can be identified by CTLs, indicating that the HLA system codes for these determinants either through gene products of a locus closely linked to or identical with the HLA-DR locus. PMID- 12735321 TI - HLA and urethritis. PMID- 12735322 TI - MNSs antigens and graft versus host disease following bone marrow transplantation. PMID- 12735324 TI - Hemangioma and HLA-B40 antigen. PMID- 12735323 TI - HLA-A and B typing in the Sarakole population of West Africa. AB - The distribution of HLA genes and haplotype frequencies was studied in a population of 119 Sarakole West Africans. The absence of HLA-B15, the rarity of HLA-A10, B12 and B17 and the relatively high frequency of HLA-A2 and B5 were noted. The presence of a Da 6 gene in the African populations is confirmed. The main gametic associations were Aw19-Bw35 and Aw23-B5. PMID- 12735325 TI - HLA and idiopathic thrombopenic purpura (ITP). PMID- 12735326 TI - Possible mechanisms by which alloantisera inhibit in the MLC test. AB - MLC inhibition studies were performed with two human alloantisera: one specific for HLA-B7, the other for HLA-DRw7. The stimulator cell inhibitory effects of these sera were tested in primary and secondary MLC tests. Both sera inhibited in the primary MLC, whereas only the anti-DRw7 serum was capable of blocking the secondary MLC test. The difference in inhibiting properties of these sera was further analyzed in primary MLC tests using selected MLC combinations, Fc receptor negative cell populations and pepsin digests of the anti-B7 serum. Anti DRw7 antibodies could inhibit by masking the stimulatory DR antigens. The inhibition of the anit-B7 antiserum was dependent on Fc, which suggested that anti HLA-B antibodies inhibited by some other mechanism. This inhibition could have been caused by antibody dependent cellular lympholysis of the stimulator cells or by the induction of suppressor cell activity. PMID- 12735327 TI - HLA-a, B typing in Basque and other Pyrenean populations. AB - Fourteen HLA-A and 18 HLA-B antigens were studied in three samples of Pyrenean populations: 198 unrelated individuals of a "Pays Basque" group; 212 non-Basque individuals from a valley in Bearn, l'Ouzom; and 73 non-Basque individuals from the neighboring valley of Bareges. The results in the Basque and the non-Basque people from l'Ouzom were comparable: the gene frequencies of HLA-A29, Aw19.2, B17 were increased and the haplotypes HLA-Aw19.2, B18; A29, B12; A2, B5; A1, B17 were found frequently with a striking linkage disequilibrium; HLA-B18 had an increased gene frequency in all these Pyrenean populations, while Bw35 was frequent in l'Ouzom and Bareges, but not among the Basques. The characteristics of Bareges were very different: the gene frequencies of HLA-A2, A11, B7 were increased while the frequency of HLA-B5 was low; the most characteristic haplotypes were HLA-A2, B12; A2, B18; A11, Bw35; A11, B27. It is interesting to note discrepancies between ethnic and HLA classification of the Basques and the non-Basque population of l'Ouzom. The HLA characteristics are quite different in the Hareges sample, more closely resembling those of Northern Europe. PMID- 12735328 TI - Antibodies against donor antigens on endothelial cells and monocytes in eluates of rejected kidney allografts. AB - Acidic eluates from two rejected kidney allografts contained antibodies reacting only with the endothelial cells and the monocytes of the specific kidney donors. No reaction was observed with the B-cells or the T-cells of these donors. When these eluates were tested against the leukocytes of healthy blood donors that were typed for HLA-A, -B, -C and DR, positive reactions were observed with the monocyte but not with the lymphocyte fraction. The first eluate reacted with 6.4% of the donors and the second eluate with 38.3%. No correlation with any of the HLA-A, -B, -C or DR antigens was found. If a more extended study proves that the endothelial-monocyte antigens do play an important role in kidney transplantation, matching of donors and recipients for these antigens and cross matching recipient serum with donor monocytes may become a necessity in the future. PMID- 12735329 TI - Further evidence for the partition of HLA-B14. AB - The minimal sequential absorption of 12 HLA-B14 antisera with B14 positive cells provided 10 antisera that showed a high correlation (r values > 0.8) with the original B14.1 and B14.2 sera of Dewar et al. (1977). The subdivisions defined by the absorbed sera corresponded to the B14 subtype of their immunizing cell. The subtyping of 112 B14 positive individuals showed the frequency of B14.1 to be 33.04%. B14 heterozygous individuals always typed for one but never for both subdivisions. Analysis of a population of 2,044 showed B14.1 to be in linkage disequilibrium with Aw32 and B14.2 with Aw33, Aw30 and A3. It is concluded that B14.1 and B14.2 are distinct antigens which, in the population tested, comprise the entire specificity of HLA-B14. PMID- 12735331 TI - Secondary responses of alloantigen-primed dog lymphocytes. AB - The usefulness of Primed Lymphocyte Typing (PLT) for recognizing Lymphocyte Defined (LD) determinants of the Major Histocompatibility Complex of the dog (DLA) was assessed in 10 separate experiments. Application of a technique described for human cell to peripheral blood lymphocytes of dogs gave reproducible, informative results. DLA LD determinants were shown to be of major importance in secondary responses of alloantigen "primed" lymphocytes, but other, a yet undefined, factors also appeared to play a role in the assay. PMID- 12735332 TI - HC restricted dual specific inhibition of mixed leukocyte culture reactions by human HLA antibody molecules. AB - A human alloantiserum was found which selectively inhibits responding cells in mixed leukocyte culture reactions. Inhibition was achieved by pre-incubation of responder cells in the antiserum followed by washing. The serum showed dual specificity as an inhibiting agent. First, inhibition was restricted to HLA-B7 or -B40 positive stimulator cells, specificities against which the antiserum also had cytotoxic activity. Second, inhibition was almost exclusively associated with the presence of the phenotype HLA-A1, -B8 on the responder cells The HLA associated specificity for responder cells was unexpected since no alloantibody activity directed to responder alloantigens could be detected by conventional serological-methods. The antiserum donor had not been immunized with HLA-A1, -B8 antigens nor with known crossreactive antigens. Furthermore, the serum donor did not carry HLA-A1, -B8 antigens herself. The inhibiting substance in the antiserum had physicochemical properties of IgG and was specifically reactive with HLA-B7 positive platelets. Pepsin digest preparations were not inhibitory. Fc receptor positive responder cells were required for inhibition. Responder cells, preincubated with the antiserum, suppressed the response of cells not incubated with the antiserum. Three possible explanations of these results are discussed: specific binding of the Fc part of the antibody with Fc receptors of responder cells, specific activation of suppressor cells and cross-reactivity. PMID- 12735330 TI - The blocking effect of antibodies against the products of the H-2 gene complex on lymphocyte complement (C3d) receptors. complement dependence and specificity. AB - Antisera against the products of the major histocompatibility system (MHS), produced by immunization between congenic mouse strains differing only at that segment of the 17th chromosome rosettes which bears the H-2gene complex, exert a reproducible blocking effect on formation of with EACm (C3d rosettes). In many instances this effect is probably due to specific antibodies reactive with the lymphocytes, as indicated by immunochemical and adsorption experiments. Purified antibodies from the C3H anti-C3H.B10 immune ascites failed to exert this blocking effect. The blocking capacity can be restored, however, by addition of fresh normal mouse, rabbit or fetal calf serum, but not by heat inactivated serum. These experiments show that the presently defined H-2K, H-2D and H-2L antigens as well as some Ia antigens are themselves not the C3d receptors. The blocking effects observed in the presence of complement are possibly due to the proximity of H-2 or Ia molecules and C3d receptors, or to a rearrangement of membrane components after reaction with anti-MHC anti-MHC antibodies and complement. PMID- 12735333 TI - HLA antigens in geographic tongue. AB - HLA-A,B,C phenotyping was performed on 95 patients with geographic tongue to determine whether there is an increased frequency of any particular allele in this condition. An increased frequency of B15 was found in the patients when compared to normal controls. When the patients were divided into atopic and non atopic groups there was an increased frequency of B15 and a decreased frequency of B40 in the atopic group compared to the controls. B40 was as decreased in the atopic group when compared to the non-atopic group. When the type I correction factor was applied to the probability values the differences in antigen frequencies in all cases became insignificant. PMID- 12735335 TI - Lack of association between the HLA-A10 (A25), B18 and C2o haplotype and anaphylactoid purpura (AP). AB - In 41 patients with AP and 134 healthy relatives HLA haplotypes and total hemolytic complement and C2 concentrations were determined. Though complement levels were found in the lower normal range in none of the individuals studied, a genetically determined C2 deficient state could be established. No association between low C2 levels and the HLA haplotype A10(A25), B18 or an isolated A25 or B18 was observed. A comparison of phenotype frequencies of AP patients with controls showed no significant difference. Contingency table analysis of patients and control haplotypes showed a definite close association with HLA-A1, Bw22, A2, Bw16, and A29, B12. PMID- 12735334 TI - Expression of H-Y antigen on preimplantation mouse embryos. AB - The expression of H-Y antigen on preimplantation mouse embryos has been studied by complement dependent cytotoxicity. Embryos at the 8-cell stage were exposed to anti-H-Y and complement and then scored for cell death. Overall, half of the treated embryos were killed, retarded, or contained dead cells. Of those embryos which were not affected by anti-H-Y, 92% were female. In control experiments, neither normal serum plus complement nor complement alone had any specific effect on male embryos. It is concluded, therefore, that the H-Y antigen is present on preimplantation mouse embryos. PMID- 12735336 TI - The first example of an HLA-Bw45 antiserum produced in a HLA-Bw44 positive woman. AB - An HLA-Bw45 serum (Mu46), which was produced by pregnancy in a Bw44 positive woman, is described. The serum was unable to be absorbed by Bw44 positive cells. Blocking tests employing broad B12 (Bw44 + Bw45) antisera, stimulated by either Bw44 or Bw45, showed that only F(ab)'2 from Bw45-stimulated B12 sera would completely block the cytotoxicity of this serum. It was concluded that Bw44 and Bw45 are distinct specificities which share common determinants. These findings are briefly discussed in relation to a recent report on the HLA-B antigen and supertypic Bw4, Bw6 antigen relationship. PMID- 12735337 TI - HLA-B8 in Raynaud's phenomenon. AB - A study of the distribution of histocompatibility (HLA) antigens in patients with Raynaud's phenomenon (RP) is presented, based upon the analysis of RP patients with and without connective tissue disease (CTD). In all patients, the phenomenon preceded the other symptoms of CTD by several years. The RP patients with CTD showed a significantly increased frequency (59%) of HLA-B8 compared with the frequency (23%) of that same antigen in the Dutch population. The RP patients without CTD showed no significant difference in frequency of HLA-B8. At least 50% of the patients with RP and HLA-B8 develop a CTD versus 18% of the patients without HLA-B8. It is suggested that HLA testing in patients with isolated RP may help to distinguish the patient in whom RP is the first symptom of CTD from the patient with truly primary RP. PMID- 12735338 TI - HLA-linked genetic control in natural rubella infection. AB - HLA antigen frequencies were investigated in 115 patients with congenital rubella syndrome and 93 of their mothers in the Ryukyu Islands, Japan. A high frequency of HLA-B15 and a low frequency of HLA-Bw22 were found in the mothers. HLA-B15 was associated with high antibody responsiveness to rubella vaccine in our previous study, whereas HLA-Bw22 was one of the HLA antigens which showed low antibody responsiveness. There was no association between HLA antigens and viral replication in rubella vaccine recipients in the present study. These results indicate that and HLA-linked genetic background in the mothers is related to susceptibility to congenital rubella syndrome, and that the mechanism of the relation may be mediated via immune response genes. PMID- 12735339 TI - Association of HLA-B12 with multiple sclerosis in India. AB - In Indian patients with Multiple Sclerosis it was observed, upon testing for serum determinable histocompatibility antigens, that HLA-B12 antigen was present in excess (77.7%), in comparison with normal controls (13.8%). In fact, in the sub-group of 'clinically definite' patients, the B12 antigen excess was remarkable (84%). These preliminary findings seem to point to a different immunogenetic profile of Indian MS patients in comparison with Western and Japanese series. Relevant published information relating to the HLA-B12 alleles in disease processes is discussed in order to provide a basis for further work. PMID- 12735340 TI - The HLA system in the Sydney Jewish population. PMID- 12735341 TI - A study of genetic markers in a family with polyendocrine disease. PMID- 12735342 TI - Detection of bovine torovirus and other enteric pathogens in feces from diarrhea cases in cattle. AB - The objectives of this study were to determine the prevalence of bovine torovirus (BoTV) in bovine fecal samples from diarrhea cases submitted to the Ohio Animal Disease Diagnostic Laboratory (ADDL) and to assess if a relationship exists between BoTV and the other enteric pathogens detected. From November 1999 to May 2001, 259 specimens from 53 calves (< or = 6 months old), 27 young adults (52 years), 125 adults (> or = 2 years), and 54 animals of unknown age were examined by an antigen-capture enzyme-linked immunosorbent assay (ELISA) and reverse transcriptase-polymerase chain reaction (RT-PCR) assay developed to detect BoTV. Testing for other enteric pathogens was performed by ADDL, and the results were analyzed with the BoTV data. The BoTV was detected using ELISA or RT-PCR in 9.7% (25/259) of the clinical samples, 56% (14/25) of which were from calves (P < 0.001) representing 26.4% (14/53) of the calves tested. Of the BoTV-positive calves, 71% (10/14) were less than 3 weeks of age. In 11/25 positive specimens, BoTV was the only pathogen detected among those examined. Other enteric organisms detected alone or in combination with BoTV in calf samples were rotavirus, coronavirus, Salmonella spp., Cryptosporidium spp., and Giardia spp.; but no consistent association between BoTV and these organisms was observed. In summary, BoTV was detected in fecal samples from cattle with diarrhea, principally in young calves less than 3 weeks of age. Future studies of infectious diarrhea in cattle should also include assays for this etiologic agent. PMID- 12735343 TI - A canine distemper outbreak in Alaska: diagnosis and strain characterization using sequence analysis. AB - Vaccination with modified-live vaccines has been very effective in reducing the incidence of canine distemper, a disease that can be devastating in unvaccinated populations. A diagnostic submission to the Animal Health Diagnostic Laboratory at Michigan State University, East Lansing, Michigan, involved a case in which several hundred dogs in an Alaskan town died in a suspected canine distemper outbreak. Cytoplasmic and intranuclear eosinophilic inclusion bodies, consistent with canine distemper virus (CDV) infection, were found in urinary bladder, spleen, lung, and salivary gland. Direct fluorescent antibody test gave results that could be considered positive for canine distemper. Because of the condition of the tissues received, the histopathology and fluorescent antibody-staining results were suggestive but not conclusive of CDV. In this study, immunohistochemistry, reverse transcriptase-polymerase chain reaction (RT-PCR), and DNA sequencing were used to confirm the presence of canine distemper virus in these tissues and to perform molecular characterization of the virus. Immunohistochemistry showed the presence of the virus in spleen, lung, and salivary gland. Viral RNA was detected by RT-PCR in brain, spleen, liver, lung, and kidney, both with nucleoprotein and phosphoprotein (P)-gene-specific primers. Sequence alignment and phylogenetic analysis of a 540-bp P-gene fragment of the Alaskan strain with corresponding sequences of 2 vaccine and 7 wild-type CDV strains showed that the virus responsible for the outbreak was closely related to a virulent strain of distemper virus from Siberia. PMID- 12735344 TI - Distribution of viral antigen and development of lesions after experimental infection of calves with a BVDV 2 strain of low virulence. AB - To examine the virus-host interaction in subclinical bovine viral diarrhea virus (BVDV) infections, the spread of a BVDV 2 strain of low virulence to different organs and the development of lesions were investigated. Eight colostrum deprived, clinically healthy, 2-3-month-old calves were intranasally inoculated with 10(6) tissue culture infective dose of the naturally occurring BVDV 2 strain 28508-5 of low virulence, and 2 served as controls. Two calves each were euthanized at days 3, 6, 9, and 13 postinoculation (pi). Representative tissues were processed for histology and immunohistology. Signs of overt clinical disease were absent. However, a mild temperature elevation at days 7 or 8 pi and a moderate decrease of circulating lymphocytes occurred in all inoculated calves. The BVDV antigen was detected at day 3 pi in several lymphoid tissues. At day 6 pi, BVDV antigen was found widespread in lymphoid tissues and multifocally in intestinal epithelial cells but was associated with no or subtle lesions only. At day 9 pi, much less BVDV antigen was detectable, but there was severe depletion of lymphoid tissues. At day 13 pi, BVDV antigen had been cleared from most lymphoid tissues that were at variable phases of depletion and recovery. In conclusion, the BVDV strain of low virulence spread to lymphoid tissues and intestinal epithelial cells but was rapidly eliminated. Transient depletion of lymphoid tissues was followed by recovery. PMID- 12735345 TI - Pooled fecal culture sampling for Mycobacterium avium subsp. paratuberculosis at different herd sizes and prevalence. AB - A stochastic spreadsheet model was developed to obtain estimates of the costs of whole herd testing on dairy farms for Mycobacterium avium subsp. paratuberculosis (Map) with pooled fecal samples. The optimal pool size was investigated for 2 scenarios, prevalence (a low-prevalence herd [< or = 5%] and a high-prevalence herd [> 5%]) and for different herd sizes (100-, 250-, 500- and 1,000-cow herds). All adult animals in the herd were sampled, and the samples of the individuals were divided into equal sized pools. When a pool tested positive, the manure samples of the animals in the pool were tested individually. The individual samples from a negative pool were assumed negative and not tested individually. Distributions were used to model the uncertainty about the sensitivity of the fecal culture at farm level and Map prevalence. The model randomly allocated a disease status to the cows (not shedding, low Map shedder, moderate Map shedder, and heavy Map shedder) on the basis of the expected prevalence in the herd. Pooling was not efficient in 100-cow and 250-cow herds with low prevalence because the probability to detect a map infection in these herds became poor (53% and 88%) when samples were pooled. When samples were pooled in larger herds, the probability to detect at least 1 (moderate to heavy) shedder was > 90%. The cost reduction as a result of pooling varied from 43% in a 100-cow herd with a high prevalence to 71% in a 1,000-cow herd with a low prevalence. The optimal pool size increased with increasing herd size and varied from 3 for a 500-cow herd with a low prevalence to 5 for a 1,000-cow herd with a high prevalence. PMID- 12735346 TI - Isolation and association of Escherichia coli AIDA-I/STb, rather than EAST1 pathotype, with diarrhea in piglets and antibiotic sensitivity of isolates. AB - To identify emerging Escherichia coli that have the potential to cause diarrhea in pigs, the prevalence of E. coli pathotypes was determined among 170 and 120 isolates from diarrheic and nondiarrheic piglets, respectively. The isolates were tested for F4, F5, F6, F18, and F41 fimbriae, for E. coli attaching and effacing (EAE), porcine attaching and effacing-associated (Paa), and adhesin involved in diffuse adherence (AIDA-I) factors, for LT, STa, STb, and enteroaggregative heat stable (EAST1) enterotoxins, and for Shiga toxins (Stxl, Stx2, and Stx2e), using DNA hybridization and polymerase chain reaction. All isolates were O-serotyped and tested for antibiotic resistance against 10 drugs. Seventeen different pathotypes, accounting for 40.0% of the isolates, were recovered from diarrheic piglets. The main pathotypes included EAST1 (13.5%), F4/LT/STb/EAST1 (6.5%), AIDA I/STb/EAST1 (4.1%), F5/STa (2.9%), EAE/EAST1 (2.9%), and AIDA-I/F18 (2.3%). Only 3 pathotypes, EAE (11.7%), EAST1 (10.8%), and EAE/EAST1 (3.3%), were recovered from nondiarrheic piglets. Paa factor was detected in 8.8% and 7.5% of isolates from diarrheic and nondiarrheic piglets, respectively, and always was associated with other virulence determinants. Overall, 22.9% of isolates from diarrheic piglets appeared to be enteropathogens: enterotoxigenic E. coli (11.7%), enteropathogenic E. coli (3.5%), and E. coli isolates (3.0%) for which none of the above adherence factors was detected. Pathotypes AIDA-I/STb/EAST1 and AIDA I/STb were isolated only from diarrheic piglets and accounted for 4.7% of isolates. Strains of these pathotypes induced diarrhea when inoculated into newborn colostrum-deprived pigs, in contrast to an isolate positive only for EAST1, which did not induce diarrhea. Antibiotic sensitivity test showed that isolates of the AIDA-I/STb/EAST1 and AIDA-I/STb pathotypes were the only strains sensitive to enrofloxacin, gentamicin, neomycin, and trimethoprim sulfamethoxazole. This study showed that at least 20.5% of isolates from diarrheic piglets appeared to be associated with AIDA-I/STb pathotype and that EAST1 pathotype is probably not an important marker for diarrhea in piglets. PMID- 12735347 TI - Protein G binding to enriched serum immunoglobulin from nondomestic hoofstock species. AB - Quick and cost-effective serologic assays, such as those based on enzyme-linked immunosorbent assay (ELISA) technology, are useful for screening animal populations for infectious diseases. Recombinant protein G is described as an almost universal ELISA conjugate for the detection of antibodies from a wide range of animal species. However, there is limited data documenting the ability of protein G to bind immunoglobulin (Ig) from many captive and free-ranging nondomestic hoofstock (Order Artiodactyla, e.g., elk, antelope, bison). Protein G binding to Ig from 11 species within this taxonomic order (addax, antelope, bison, bontebok, elk, impala, kudu/nyala, muntjac, oryx, sheep, and white-tailed deer) and 2 control species (bovine and chicken) was assessed. A serum Ig enrichment protocol, using high-performance liquid chromatography (HPLC), was optimized in bovids (Bos taurus) and then applied to the other study species. Binding assays were performed by adding protein G to microtiter wells coated with titrated dilutions of enriched artiodactyl Ig. Optical densities were measured and binding curves generated. Differences in protein G binding were observed, both within and among species, as well as within taxonomic families. Significant intraspecies binding variation was observed for 7 species tested (antelope, oryx, sheep, muntjac, impala, bontebok, and addax). No statistically significant intraspecies differences in protein G binding were found for Ig from bison, elk, kudu/nyala, white-tailed deer, plus control species (cattle and chicken). Binding of protein G to Ig from impala, muntjac, and elk was statistically different from the positive control (cattle), with muntjac binding curves statistically comparable with the negative control (chicken). For the other 7 species tested, binding curves illustrated the ability of protein G to bind Ig as well as, or better than, the positive control. These findings expand the list of animal species whose Ig is capable of being detected using recombinant protein G, with the caveat that protein G does not bind Ig uniformly in closely related species. It is concluded that recombinant protein G conjugates may serve as useful reagents for serodiagnosis by ELISA in nondomestic hoofstock, although different assay interpretation algorithms and assay protocols may need to be developed on a per species basis for maximum diagnostic effectiveness. PMID- 12735348 TI - A novel multiplex polymerase chain reaction approach for detection of four human infective Cryptosporidium isolates: Cryptosporidium parvum, types H and C, Cryptosporidium canis, and Cryptosporidium felis in fecal and soil samples. AB - A nested multiplex polymerase chain reaction (PCR) approach was adopted for the simultaneous detection of 4 human infective genotypes of the protozoan parasite Cryptosporidium. Specific PCR primers were designed for the heat shock protein 70 gene of 2 genotypes of Cryptosporidium parvum (human and bovine types), Cryptosporidium canis, and Cryptosporidium felis. These 4 genotypes have all been found in human fecal samples. The primers amplified DNA fragments of specific sizes, each representing a unique genotype. The limit of detection of the method was found to vary between 10 and 100 oocysts per 1 ml fecal material. There appeared to be no cross-reactivity with other organisms commonly present in feces and soil, and the approach has a high specificity. The rapid identification of various human infective Cryptosporidium isolates is a part of the authors' long term aim of determining the routes of infection with oocysts and thereby increase their epidemiological understanding of Cryptosporidium infection in humans and animals. PMID- 12735349 TI - The use of a mimic to detect polymerase chain reaction-inhibitory factors in feces examined for the presence of Lawsonia intracellularis. AB - Lawsonia intracellularis is an intracellular organism that causes proliferative enteritis in pigs. This bacterium is difficult to culture, and antemortem demonstration of the microbe is therefore often performed on fecal samples by polymerase chain reaction (PCR). Polymerase chain reaction is sensitive and specific, but inhibitory factors in feces might cause false-negative results. This article describes the construction and use of an internal standard, a mimic. The mimic is amplified by the same primers as those used for L. intracellularis DNA and thus could indicate false-negative results in clinical samples. The amplicon was clearly visible when as few as 10 mimic molecules were added per amplification reaction and when no inhibitors werepresent. When fecal samples were spiked with the mimic, the detection limit was 10(2) molecules per PCR. Sixty clinical samples, 20 from wild boars, 20 from growing pigs with diarrhea, and 20 from pigs without diarrhea, were prepared by a boiling procedure and subjected to PCR together with 10(3) mimic molecules. Nine samples were positive, of which 7 originated from pigs with diarrhea and 2 from pigs without diarrhea. In 14 samples from wild boars, in 8 samples from pigs without diarrhea, and in 3 samples from pigs with diarrhea, neither the mimic nor the target DNA was visible. This indicated the presence of inhibitors in these samples. It is concluded that the mimic can be used as an internal control in the diagnosis of L. intracellularis to indicate inhibition of PCR. PMID- 12735350 TI - Survey of cattle in northeast Colorado for evidence of chronic wasting disease: geographical and high-risk targeted sample. AB - A geographically targeted survey of potentially high-risk, adult cattle in chronic wasting disease (CWD)-endemic areas in Colorado was initiated to assess the possibility of the spread of CWD from deer to cattle under natural conditions. Surveyed cattle were sympatric with free-roaming deer in geographically defined areas where CWD occurs and where CWD prevalence has been estimated. To qualify for inclusion in the survey, cattle had to be at least 4 years old and had to have spent a minimum of 4 years in surveyed areas. Brains from culled cattle were examined microscopically and immunohistochemically for tissue alterations indicative of a transmissible spongiform encephalopathy (TSE). Two hundred sixty-two brains were suitable for evaluation and were found to lack changes indicative of a TSE infection. Prion deposition was not demonstrable using a method involving formic acid and proteinase-K treatment before application of monoclonal antibody to bovine prion protein (F99/97.6.1). Some incidental neuropathologic changes unrelated to those of TSEs were detected. Findings from this study suggest that large-scale spread of CWD from deer to cattle under natural range conditions in CWD-endemic areas of northeast Colorado is unlikely. PMID- 12735351 TI - Comparison of targeting F and G protein genes to detect bovine and ovine respiratory syncytial viruses. AB - In this study, 2 reverse transcriptase-polymerase chain reaction (RT-PCR) assays were developed and compared for simultaneous detection of bovine and ovine respiratory syncytial viruses (RSVs). One assay was based on a set of primers, which amplified a 426-bp fragment of either bovine or ovine RSV F gene (RT-PCR F). The F products could be distinguished by EcoRI or BstYI restriction endonuclease cleavage. In the other assay, a set of primers amplified a 542-bp fragment of either ovine or bovine RSV G gene (RT-PCR G). EcoO1091 and RsaI restriction enzymes were used to differentiate between the ovine and bovine PCR-G products. Sequencing of the PCR products confirmed the fidelity of both assays. The 2 assays were evaluated using 18 bovine RSV isolates, 1 ovine RSV, 1 bighorn sheep RSV isolate, 1 caprine RSV isolate, 2 human RSV isolates, and several other viruses associated with bovine respiratory tract disease. RT-PCR G may be more sensitive in detecting viral RNA. Because the target sequence of the F gene is more conserved than that of the G gene, RT-PCR F followed by the appropriate restriction enzyme cleavage may be superior to RT-PCR G to discriminate between the 2 ruminant RSV subgroups. This assay should prove useful for determining the relative contribution of ovine and bovine RSV to the pathogenesis of bovine respiratory tract disease. PMID- 12735352 TI - Comparison of three enzyme-linked immunosorbent assays for serologic diagnosis of contagious agalactia in sheep. AB - Serologic diagnosis of ovine contagious agalactia (Mycoplasma agalactiae) with the enzyme-linked immunosorbent assay (ELISA) developed by Agence Francaise de Securite Sanitaire des Aliments (AFSSA) may produce a few false-positive (FP) and false-negative (FN) results. When the prevalence of disease is low, these erroneous results may generate problems for eradication schemes. To prevent this, 2 commercial ELISAs were compared with the AFSSA ELISA. Flocks of known status were selected and classified into 4 categories: true positive (TP), FP, true negative (TN), and FN; 20 sheep per flock were submitted for blood sampling. A flock was considered positive when at least 1 out of 20 sera was positive or 2 sera were doubtful. In the flock, the diagnostic sensitivity of the 3 kits was very good (100%), and the diagnostic specificity showed an improvement from 46% (AFSSA test) to 88% and 92% (commercial tests). Considering individual animals, very few positive ewes were detected within TN or FP flocks; the proportion of positive ewes varied greatly from one kit to another (48% to 82%) within TP flocks. The kinetics of antibody response in sheep experimentally infected with various field strains of M. agalactiae were quite similar with all 3 ELISAs. The agreement between the 3 tests, assessed using the kappa value, varied from moderate to good (respective values of 0.56, 0.61, and 0.86). The 2 commercial ELISAs showed better performances, probably because of a superior analytical sensitivity, and are a good alternative for the serodiagnosis of contagious agalactia in sheep. PMID- 12735353 TI - Coccygeal chordoma in a dog. AB - A 4-cm diameter spherical mass was removed from the region of the second coccygeal vertebra of a 6-year-old female Doberman Pinscher dog. The mass had been present for 7 months and was described as multilobular and smooth. Histologically, the neoplasm consisted of small spindle-shaped cells surrounding lobules of large vacuolated polygonal cells. Mucin was present within most lobules. The large polygonal cells stained positively with antibodies against vimentin, cytokeratin, and S-100 protein. Although the spindle-shaped cells stained positively with antibodies against vimentin, they only occasionally contained cytokeratin. The histology and immunohistochemistry are consistent with that reported for chordoma. Although 4 chordomas have previously been reported in dogs, this is the first case in which immunohistochemistry has been used to support the histological diagnosis. Neither recurrence nor metastasis was reported 10 months after surgical excision. PMID- 12735354 TI - Lightning injury in an outdoor swine herd. AB - Three pigs, weighing 63 kg-70 kg each, from a group of 8 pigs in an outdoor pen that was struck by lightning were necropsied. All 3 pigs presented with hind limb paralysis. The only lesions identified were multiple fractures of the last (seventh) lumbar vertebral body and first sacral vertebral segment, with dorsal displacement of the sacrum and transection of the distal spinal cord and spinal nerves. Hemorrhages extended from the fracture sites into muscles immediately surrounding the lumbosacral junction and retroperitoneally into the pelvic cavity. These hemorrhages were not clearly visible until the pelvic region was dissected. Lesions commonly found in human lightning-strike victims were not present in these pigs. Because vertebral fractures may be the only lesions and may be grossly subtle in heavily muscled pigs, careful pelvic and vertebral dissection is recommended in cases of suspected lightning strike and electrocution. PMID- 12735355 TI - Pulmonary sarcocystosis in a puppy with canine distemper in Costa Rica. AB - Canine distemper and pulmonary sarcocystosis were diagnosed in a 10-week-old Rottweiler with 4-day history of diarrhea, vomiting, and weakness. Microscopic examination of the lung revealed bronchointerstitial pneumonia typical of morbillivirus infection. Also, numerous apicomplexan parasites were scattered in the alveolar walls. This protozoan infection was first thought to be toxoplasmosis but immunoperoxidase staining revealed large numbers of Sarcocystis canis. This is the first case of canine sarcocystosis reported from Latin America that further emphasizes the importance of immunohistochemistry in the differential diagnoses of apicomplexan infections in dogs. PMID- 12735356 TI - Descriptive epidemiology of late-term abortions associated with the mare reproductive loss syndrome in central Kentucky. AB - Epidemiological and pathological findings of 433 late-term abortions associated with the mare reproductive loss syndrome (MRLS) in central Kentucky were identified by reviewing the records of the University of Kentucky Livestock Diseases Diagnostic Center. The distribution of dates of abortion was clustered during a brief period of time, presumably from a simultaneous environmental exposure. The most common pathological findings were microscopic pulmonary lesions consisting of squamous epithelial cells present in alveoli with or without concurrent infiltration of inflammatory cells (neutrophils, macrophages, or monocytes) in the interstitium or within alveoli. Isolation of a non-beta hemolytic Streptococcus (52% of fetuses) or an Actinobacillus sp. (19% of fetuses) was common. Placentitis or funisitis was identified in 44% of fetuses. No single pathological finding, however, was pathognomonic for MRLS-associated late-term abortion. This report describes the pathological findings characterizing the MRLS-associated abortion. A cause of MRLS could not be determined from necropsy findings. PMID- 12735357 TI - Citrobacter freundii septicemia in two dogs. AB - A 4-month-old Maltese puppy and a 7.5-year-old Collie were diagnosed with septicemia associated with Citrobacter freundii. The puppy died soon, after developing weakness and mucohemorragic diarrhea. The Collie had immune-mediated hemolytic anemia and thrombocytopenia and was treated with immunosuppressive drugs before being euthanized. Gross examination of the puppy revealed mucohemorrhagic intestinal contents. Focal necrotic hepatitis, fibrinous peritonitis, interstitial pneumonia, and hemorrhagic gastrointestinal contents were observed in the older dog. Histologically, there was a diffuse, moderate, histiocytic meningitis in the puppy and a focal fibrinonecrotic hepatitis in the adult dog. Lesions in both dogs contained numerous gram-negative rods. Citrobacter freudii is a potential cause of monomicrobic bacteraemia-septicemia in puppies or immunocompromized adult dogs. The gastrointestinal tract is probably the main site of entry. PMID- 12735358 TI - Unconditioned sexual incentive motivation in the male Norway rat (Rattus norvegicus). AB - Sexual incentive motivation was evaluated in a procedure consisting of a large open field where incentive animals were confined behind wire mesh openings. When sexually inexperienced male rats (Rattus norvegicus) were exposed to the receptive female-male incentives, they spent more time close to the female. If the incentives were receptive female-nonreceptive female, the receptive female was preferred. However, when the alternatives were nonreceptive female-male, no preference was obtained. Castration abolished preference for the receptive female, and treatment with testosterone propionate restored it. Estradiol plus oil is as efficient as estradiol plus progesterone for giving the ovariectomized female incentive properties. The living female can be replaced with female odor. Sexual experience did not have any long-term effects on the female's incentive value, but immediately preceding limited sexual activity enhanced it PMID- 12735359 TI - Global and local processing of hierarchical visual stimuli in tufted capuchin monkeys (Cebus apella). AB - Capuchin monkeys' (Cebas apella) relative accuracy in the processing of the global shape or the local features of hierarchical visual stimuli was assessed. Three experiments are presented featuring manipulations of the arrangement and the density of the local elements of the stimuli. The results showed a clear advantage for local level processing in this species, which is robust under manipulations of the density of the local elements of the stimuli. By contrast the density of the component elements linearly affected accuracy in global processing. These findings, which support those from other studies in which a local superiority emerged in animals, challenge the generality of early claims concerning the adaptive value of global advantage in the processing of hierarchical visual patterns. PMID- 12735360 TI - Lower begging responsiveness of host versus parasitic brown-headed cowbird (Molothrus ater) nestlings is related to species identity but not to early social experience. AB - The survival of young brood parasites depends critically on their many adaptations to exploit hosts. Parasitic survival is particularly related to competitive superiorty for foster parental care whenever host young are not destroyed in parasitized nests.Brown-headed cowbirds (Molothrus ater) are generalist obligate parasites whose early social environments are unpredictable regarding host species and numbers of nestmates. Young avian brood parasites typically beg more intensively and loudly than foster siblings, but an untested prediction is that young parasites are also more likely to respond by begging to a wider variety of stimulus types. Avian vocalizations were used in a playback experiment to stimulate begging behavior in cowbird hosts. Compared with age matched cowbird nestlings, hosts begged less frequently to acoustic stimuli, and lower begging responsiveness was irrespective of whether hosts had been reared in parasitized nests. PMID- 12735361 TI - Delayed alternation in honeybees (Apis mellifera). AB - Previous experiments with honeybees (Apis mellifera) failed to show learned control of performance by short-ten memory. In this study, honeybees were trained with an improved technique to choose 1 of 2 colors that was either the same as a recently rewarded sample (perseveration) or different (alteration). Because any increase in associative strength stemming from the sample experience would tend to promote perseveration and contravene alternation, the equal difficulty of the 2 tasks suggests that the role played by the sample was primarily discriminative. The animals remembered on each trial the immediately preceding experience with reward and learned to use that information appropriately. These new results extend the list of what may well be fundamental similarities in the learning of vertebrates and honeybees. PMID- 12735362 TI - Ontogeny of social behavior in the megapode Australian brush-turkey (Alectura lathami). AB - Megapodes meet conspecifics at an unpredictable age, and it is unknown how their social behavior develops under such conditions. The authors induced encounters between 2-day-old socially naive hatchlings and up to 49-day-old chicks of the Australian brush-turkey (Alectura lathami) in a large outdoor aviary. All social behavior patterns found in older chicks were present in hatchlings and did not change in form or the frequency in which they occurred thereafer; the frequency of calling was the only feature that changed significantly with age. Chicks stayed only 0.3-2.0 m (medians) apart and synchronized their feeding activity from the age of 20 days. These results indicate that megapode chicks show social behavior without social experience and that this requires no postnatal leaning. PMID- 12735363 TI - Classification of domestic cat (Felis catus) vocalizations by naive and experienced human listeners. AB - To test for possible functional referentiality in a common domestic cat (Felis catus) vocalization, the authors conducted 2 experiments to examine whether human participants could classify meow sounds recorded from 12 different cats in 5 behavioral contexts. In Experiment 1, participants heard singlecalls, whereas in Experiment 2, bouts of calls were presented. In both cases, classification accuracy was significantly above chance, but modestly so. Accuracy for bouts exceeded that for single calls. Overall, participants performed better in classifying individual calls if they had lived with, interacted with, and had a general affinity for cats. These results provide little evidence of referentiality suggesting instead that meows are nonspecific, somewhat negatively toned stimuli that attract attention from humans. With experience, human listeners can become more proficient at inferring positive-affect states from cat meows. PMID- 12735364 TI - Strategy planning in cats (Felis catus) in a progressive elimination task. AB - Domestic cats (Felis cans) were administered a progressive elimination task in which they had to visit and deplete 3 baited sites. Cats were brought back to the starting point after each visit to any site whether the visit represented a correct or an incorrect choice. The results revealed that cats organized search as a function of the least distance principle whether they had to assess starting point to target distances (Experiment 1, n = 12) or adjacent bowl distances (Experiment 2, n = 12). The results also revealed that the starting point to target distance factor was the most influential in determining the initial choice (Experiment 3, n = 6) and in producing errors. Errors were also linked to the antero-posterior bodily axis of the cat. Results are discussed in terms of the predatory behavior of the cat. PMID- 12735365 TI - Does zebra finch (Taeniopygia guttata) preference for the (familiar) father's song generalize to the songs of unfamiliar brothers? AB - Several studies have demonstrated that zebra finches (Taeniopygia guttata) prefer their fathers' songs over unfamiliar songs. Songs of tutors (i.e., fathers) and tutees (i.e., sons) resemble each other as a result of cultural transmission. Subjects (N = 18) with a previously established preference for the father's song could choose between the song of an unfamiliar brother or a random unfamiliar song in an operant task. Most subjects showed a significant preference for either category of song, but overall, the songs of unfamiliar brothers were not preferred, although they were more similar to the father's song than were the unfamiliar songs. This suggests that subjects did not generalize their learned preference for a song of a particular tutor to the songs of his tutees. PMID- 12735366 TI - Withholding information in semifree-ranging Tonkean macaques (Macaca tonkeana). AB - The ability of Tonkean macaques (Macaca tonkeana) to deceive partner about the location of hidden food was investigated in 4 males belonging to a group raised in a 2-acre park. Before releasing subjects in the search task, the experimenter prompted 1 of the subjects to observe where the bait was hidden. The authors found that when informed, higher ranking subjects did not significantly alter their search patterns whether tested alone or in pairs. The same held for subordinates belonging to pairs with weak dominance asymmetry. To the contrary, strongly dominated subordinates modified the number of pauses when searching in pairs. They used behavioral tactics such as avoiding being followed, stopping when being watched, or taking a wrong direction. Information withholding might be acommon event in macaques. PMID- 12735367 TI - Extinction of conditioned sexual responses in male Japanese quail (Coturnix japonica): role of species-typical cues. AB - The authors examined how a conditioned stimulus (CS) that included species typical cues affected the acquisition and extinction of conditioned sexual responses in male quail (Coturnix japonica). Some subjects were conditioned with a CS that supported sexual responses and included a taxidermic head of a female quail. Others were conditioned with a similar CS that lacked species-typical cues. Pairing the CSs with access to live females increased CS-directed behavior, with the head CS eliciting significantly more responding than the no-head CS. Responding to the head CS persisted during the 42-day, 126-trial extinction phase; responses to the no-head CS extinguished. Responding declined when the cues were removed or the subjects were sexually satiated. Possible functions and mechanisms of these effects are discussed. PMID- 12735368 TI - Hearing and vocalizations in the orange-fronted conure (Aratinga canicularis). AB - The auditory sensitivities of the orange-fronted conure (Aratinga canicularis) were examined in relation to the spectral characteristics of its vocalizations. Absolute thresholds, masked thresholds, frequency difference limens, and intensity difference limens for pure tones were obtained using psychoacoustic techniques. In general, hearing abilities are similar to those found in many avian auditory generalists. One exception is the unusually low critical ratio (masked threshold) between 2.0 and 4.0 kHz, similar to that previously found in the budgerigar (Melopsittacus undulatus). These auditory sensitivities were compared with average spectra for (a) contact calls and (b) a general sample of vocalizations recorded from wild birds. The spectral regions of both greatest vocal energy and best auditory sensitivity were between 2.0 and 5.0 kHz. PMID- 12735369 TI - Preference for novel flavors in adult Norway rats (Rattus norvegicus). AB - The authors fed rats 1 of 2 distinctively flavored, roughly equipalatable diets for 3 days then offered them an ad libitum choice between the 2 diets. For 3 days, subjects exhibited a reduced relative intake of whichever diet they had previously eaten (Experiment 1). Such reduction in relative intake was as effective as a toxicosis-induced conditioned aversion in determining subjects' food choices (Experiment 2). The strength of exposure-induced reduction in relative intake did not depend on similarity of the 2 diets offered for choice either to each other or to subjects' maintenance diet (Experiment 3) but did require continuous exposure to a diet (Experiment 4). These experiments provide the first evidence of a robust, exposure-induced decrease infood preference in rats lasting for days rather than minutes. PMID- 12735370 TI - The mother-offspring relationship as a template in social development: reconciliation in captive brown capuchins (Cebus apella). AB - Mother-offspring (MO) relationship quality was investigated to determine its influence on the development of reconciliation--affiliation between opponents shortly after a fight--because it influenceswhat distressed youngsters learn about calming down. Data were longitudinal and cross-sectional observational samples of 38 MO pairs of monkeys across 24 months. An MO relationship quality index (RQI) classified each pair as secure or insecure. Reconciliation emerged in infancy. Secure youngsters had an appeasing conciliatory style, and insecure youngsters had an agitated conciliatory style. Conclusions are that reconciliation develops from the attachment behavior system and MO RQI is related to the particular conciliatory style youngsters develop by affecting how aroused they are by conflict and the subsequent socializing they seek to calm down. PMID- 12735371 TI - Using visual reinforcement to establish stimulus control of responding of Siamese fighting fish (Betta splendens). AB - Stimulus control of ring swimming was studied with male Siamese fighting fish (Betta splendens) using 2-component multiple schedules in which the components were correlated with the presence or absence of air bubbles in the water. In Experiment 1, either response-independent mirror presentations or extinction was juxtaposed with immediate response-dependent mirror presentations. Rates of ring swimming generally were higher with immediate reinforcement than with either response-independent mirror presentations or extinction. In Experiment 2, different durations of response-dependent mirror presentations were juxtaposed. Generally, higher rates of ring swimming occurred with 15-s than with 0-, 1-, or 3-s durations. Results demonstrate that stimulus control of responding can be established with these fish under several conditions of differential reinforcement. PMID- 12735373 TI - Accuracy of arthroscopic assessment of anterior ankle cartilage lesions. AB - PURPOSE: The accuracy of arthroscopic evaluation of the size of an osteochondral lesion in the ankle joint was assessed in 10 cadaver feet. MATERIALS AND METHODS: A rectangular osteochondral defect was created in the anterior part of the talus. A 5 mm 30 degrees arthroscope was utilized for evaluation of the size of the lesion from an anterior midline portal under carbon dioxide. RESULTS: The size of the defect averaged 77.2 +/- 31 mm2 (24-10(8) mm2). The difference between area of the defect and measurement of three independent investigators averaged 52%, 49% and 49%, respectively. CONCLUSION: The assessment of the size of an osteochondral lesion in the ankle joint based on arthroscopy implicates over- and underestimation of the defect. PMID- 12735374 TI - Revision of ankle arthrodesis. AB - From 1989 to 1996, we treated 18 cases (10 males, eight females; average age 48.2 years) of failed ankle arthrodesis by revision of ankle arthrodesis and followed their progress for at least two years. The average time interval between original surgery and revision was 17.3 months. Revisions were needed due to infection in one case, nonunion in 10 cases, and malalignment in seven cases. The salvage operations included debridement in the infected case, refreshed pseudoarthrosis in nonunion cases, and corrective osteotomy in malalignment cases. Sixteen cases were fixed by crossed screws with internal compression, one infected case was fixed by an external fixator, and one case with bone loss was fixed with buttress plate. The average follow-up period was 40.4 months. There was one nonunion and two delayed unions, with an ultimate fusion rate of 94%. The average AOFAS ankle hindfoot score was 70.9 at final follow up. There was one excellent result (5.6%), five good results (27.8%), 11 fair results (61%), and one poor result (5.6%), and the overall results were poorer compared with our series of primary arthrodesis. The time to fusion also took longer in the revision cases (average 2.7 months in primary cases and 4.8 months in revision cases). Fusion techniques that ensure solid union in a functional position are essential. If an ankle arthrodesis fails, however, revision is a salvage procedure that can achieve an acceptable result. PMID- 12735375 TI - Radiographic changes in forefoot geometry with weightbearing. AB - Quality assessment of forefoot surgery depends mainly on weightbearing radiographs. A prospective study has been performed to compare the influence of weightbearing on forefoot geometry. Dorsoplantar radiographs for weightbearing and non-weightbearing conditions were performed in 99 patients. Hallux valgus angle, the intermetatarsal angles between the first and second and first and fifth metatarsals and intermetatarsal distance were measured using an interactive digitizer connected to a computer. The intermetatarsal angles showed a statistically significant increase during weightbearing. Unrelated to the severity of hallux valgus deformity, hallux valgus angles demonstrated an inverse behavior showing larger values under non-weightbearing conditions. For that reason, radiological evaluation of forefoot geometry strictly requires similar weightbearing conditions and comparable positioning of the foot. PMID- 12735376 TI - First metatarsal-phalangeal joint arthrodesis: a biomechanical assessment of stability. AB - BACKGROUND: First metatarsal phalangeal joint (MTP) arthrodesis is a commonly performed procedure for the treatment of hallux rigidus, severe and recurrent bunion deformities, rheumatoid arthritis and other less common disorders of the joint. There are different techniques of fixation of the joint to promote arthrodesis including oblique lag screw fixation, lag screw and dorsal plate fixation, crossed Kirschner wires, dorsal plate fixation alone and various types of external fixation. Ideally the fixation method should be reproducible, lead to a high rate of fusion, and have a low incidence of complications. METHODS: In the present study, we compared the strength of fixation of five commonly utilized techniques of first MTP joint arthrodesis. These were: 1. Surface excision with machined conical reaming and fixation with a 3.5 mm cortical interfragmentary lag screw. 2. Surface excision with machined conical reaming and fixation with crossed 0.062 Kirschner wires. 3. Surface excision with machined conical reaming and fixation with a 3.5 mm cortical lag screw and a four hole dorsal miniplate secured with 3.5 mm cortical screws. 4. Surface excision with machined conical reaming and fixation with a four hole dorsal miniplate secured with 3.5 mm cortical screws and no lag screw. 5. Planar surface excision and fixation with a single oblique 3.5 mm interfragmentary cortical lag screw. Testing was done on an Instron materials testing device loading the first MTP joint in dorsiflexion. Liquid metal strain gauges were placed over the joint and micromotion was detected with varying loads and cycles. RESULTS: The most stable technique was the combination of machined conical reaming and an oblique interfragmentary lag screw and dorsal plate. This was greater than two times stronger than an oblique lag screw alone. Dorsal plate alone and Kirschner wire fixation were the weakest techniques. CONCLUSIONS: First MTP fusion is a commonly performed procedure for the treatment of a variety of disorders of the first MTP joint. The most stable technique for obtaining fusion in this study was the combination of an oblique lag screw and a dorsal plate. This should lead to higher rates of arthrodesis. PMID- 12735377 TI - Malunion following trimalleolar fracture with posterolateral subluxation of the talus--reconstruction including the posterior malleolus. AB - Malunion after a malleolar fracture can include a displaced posterior malleolus with associated posterolateral subluxation of the talus. Corrective osteotomy including the posterior malleolus was performed in four patients. Joint congruity was obtained in every case. The patients were followed for 46 to 80 months postoperatively. They all experienced an improvement in pain and three of four patients were unlimited in their walking capacity. Mild to moderate residual symptoms were frequent. The symptoms were attributed to the damage of the cartilage and soft-tissues both from the initial injury and from weightbearing on the incongruous joint. Discrete, non-progressive osteophytes were seen in all patients. Delay in reconstruction did not preclude a good result, although early reoperation is felt to be preferable. PMID- 12735379 TI - The relationship of the position of the metatarsal heads and peak plantar pressure. AB - We test the premise that peak plantar pressure is located directly under the bony prominences in the forefoot region. The right foot of standing volunteers was examined in three different postures by a CT-scanner. The plantar pressure distribution was simultaneously recorded. The position of the metatarsal heads and the sesamoids could be related to the corresponding local peak plantar pressures. The metatarsal heads 1, 4, and 5 had a significantly different position than the local peak plantar pressures. The average difference in distance between the position of the metatarsal heads and the peak plantar pressure showed a significant correlation: on the medial side the head was located more distally to the local peak plantar pressure, on the lateral side more proximally. The findings suggest that normal plantar soft tissue is able to deflect a load. The observations might improve insight into the function of the normal forefoot and might direct further research on the pathological forefoot and on the design of footwear. PMID- 12735380 TI - MR morphometry of posterior tibialis muscle in adult acquired flat foot. AB - We conducted magnetic resonance imaging of the posterior tibial (PT) and flexor digitorum longus (FDL) muscle bellies in 12 patients undergoing surgical treatment for unilateral posterior tibial tendon (PTT) dysfunction. All patients had atrophy of the PT muscle compared to the normal leg (mean 10.7%, p = 0.008). In those patients with a complete rupture of PTT there was replacement of the PT muscle by fatty infiltration. Conversely, the FDL muscle showed a compensatory hypertrophy (mean 17.2%, p < 0.002). We support the use of FDL as an appropriate tendon for augmentation of PTT in stage II disease. This study also demonstrates that in the presence of a complete rupture, excision of the PTT is a reasonable surgical procedure and pure tenodesis will be more likely to fail because the PT muscle belly undergoes fatty infiltration. In patients with a diseased but intact PTT there was no fatty infiltration and the muscle volume was at least 83% of the normal side in all cases. We therefore suggest that in the presence of an intact PTT the PT muscle belly may provide some useful function if used to augment the FDL transfer when the diseased tendon is excised. PMID- 12735381 TI - A biomechanical evaluation of clinical stress tests for syndesmotic ankle instability. AB - Displacement transducers were placed across the anterior and posterior tibiofibular ligaments of 17 fresh cadaver (78.4 +/- 6.7 years old at death) lower extremities. Displacements induced by various clinical tests (squeeze, fibula translation, Cotton, external rotation, and anterior drawer) were measured with the ankle ligaments intact and after sequential sectioning of the anterior tibiofibular ligament, anterior deltoid ligament, and posterior tibiofibular ligament. None of the syndesmotic stress tests could distinguish which ligaments were sectioned. Furthermore, the small displacements measured during the stress tests (with the exception of the external rotation test) suggest it is unlikely that the displacement induced in injured syndesmoses can be clinically differentiated from normal syndesmoses. Therefore, pain, rather than increased displacement, should be considered the outcome measure of these tests. PMID- 12735378 TI - Lengthening of short great toe and correction of all lesser toe deformities by distraction-lengthening. PMID- 12735382 TI - Three-dimensional starch model for simulation of corrective osteotomy for a complex bone deformity: a case report. AB - The complex valgus deformity of the right ankle of a 24-year-old Maffucci syndrome man was corrected by three-dimensional osteotomy followed by limb lengthening. Before surgical correction of the deformity, we used computed tomography data to make a life-size three-dimensional plastic model of the deformed ankle for an accurate understanding of the anatomical deformity. We then used this model to perform a simulated osteotomy. The real osteotomy was performed immediately afterwards and valgus and recurvatum deformities were corrected accurately. We recommend simulated surgery using a three-dimensional plastic model which will improve the pre-operative planning technique and the accuracy of the end results. PMID- 12735383 TI - Primary fusion as salvage following talar neck fracture: a case report. AB - For a 29-year-old man with a three-week-old Hawkins Type IV talar neck fracture, intra-operative reduction and fixation were not possible due to soft tissue contractures and severe comminution. A primary talonavicular and subtalar arthrodesis with the use of iliac crest bone graft was performed. Postoperative follow-up at 16 months demonstrated solid fusions, no avascular necrosis of the talus and a functional range of motion at the ankle. He was not capable of returning to his job of roof maintenance. PMID- 12735384 TI - Technique tip: fixation of Mitchell's osteotomy using a PLLA screw. PMID- 12735385 TI - Spermatogenic patterns and early embryo development after intracytoplasmic sperm injection in severe oligoasthenozoospermia. AB - PURPOSE: Evaluate the influence of different baseline spermatogenic patterns [meiotic pattern (normal or abnormal), sperm concentration (> 1 x 10(6)/mL or < or = 1 x 10(6)/mL), and the combined meiosis-sperm concentration pattern] on early embryo development in severe oligoasthenozoospermia. METHODS: Embryo outcomes (fertilization rate, cleavage rate, and 4-cell stage embryo division rate on day 2) after IVF-ICSI in 75 oligoasthenozoospermia and 79 normozoospermic males. RESULTS: The embryo division rate was significantly lower in oligoasthenozoospermia compared to normozoospermia (50.43% vs. 58.72%, p < 0.01) and in the oligoasthenozoospermia group for meiotic anomalies (43.40%), sperm concentration < or = 1 x 10(6)/mL (44.35%), and the combined pattern < or = 1 x 10(6)/mL with meiotic anomalies (37.17%). Logistic regression analysis showed a synergic effect (OR = 2.00; 95% CI = 1.28-3.12) when the two spermatogenic patterns predictive of slow embryo development [meiotic anomalies (OR = 1.49; 95% CI = 1.03-2.15) and sperm concentration < or = 1 x 10(6)/mL (OR = 1.53; 95% CI = 1.09-2.13)] were present. CONCLUSIONS: The data suggest that the early embryonic developmental capacity is inversely related to the severity of spermatogenic impairment (meiotic anomalies and/or sperm concentration < or = 1 x 10(6)/mL). PMID- 12735388 TI - Cytogenetic analysis of spontaneously activated noninseminated oocytes and parthenogenetically activated failed fertilized human oocytes--implications for the use of primate parthenotes for stem cell production. AB - PURPOSE: [corrected] Spontaneous parthenogenetically activated noninseminated oocytes and failed fertilized oocytes after ART activated by puromycin were studied to assess cleavage ability and the cytogenetic constitution of the resulting embryos. METHODS: Failed fertilized oocytes were exposed to puromycin, and whenever activation occurred, they were further cultured until arrest of development. FISH was used to assess the ploidy of spontaneous (group A) and induced parthenotes (group B). RESULTS: The mean number of oocytes exposed to puromycin and the percentage and type of activation were identical in IVF and ICSI patients. The more frequent types of activation were one or two pronuclei and one polar body suggesting that retention of the second polar body is a common event after parthenogenetic activation. CONCLUSIONS: Retention of the second polar body and chromosome malsegregation were observed after parthenogenetic activation, either spontaneous or induced by puromycin. This means that using parthenogenetic embryos for stem cell research will require great care and attention. PMID- 12735389 TI - The incidence of major birth defects following in vitro fertilization. AB - PURPOSE: To evaluate the risk of congenital malformations in newborns delivered after IVF-ET in comparison with matched controls from spontaneous pregnancies. METHODS: A total of 12,920 deliveries were subjected to retrospective analysis. A total of 301 neonates were evaluated. The incidence of major birth defects was compared with controls matched with regard to age, gravidity, parity, and previous obstetric outcome after spontaneous pregnancies. RESULTS: The incidence of major congenital abnormalities was not significantly higher (p > 0.05) among the cases (1.90%) than among the controls (1.15%). CONCLUSION: The risk of major birth defects following IVF-ET is comparable with that of spontaneously conceived, matched pregnancies. PMID- 12735386 TI - Effect of ICSI on subsequent blastocyst development and pregnancy rates. AB - PURPOSE: To investigate whether ICSI (intracytoplasmic sperm injection) results in decreased blastocyst formation and pregnancy compared to IVF (in vitro fertilization). METHODS: We performed a retrospective analysis of blastocyst transfer (BT) offered routinely to patients under age 40 with > or = three 8-cell embryos on day 3 and compared IVF to ICSI cycles. Sequential media were used with P1 until day 3, then Blastocyst Medium until day 5/6. RESULTS: There were 131 IVF and 75 ICSI cycles. There was no difference in age, number of oocytes, zygotes, 8 cell embryos, blastocysts on days 5 and 6, or embryos transferred. Progression to blastocyst was similar (78% for IVF and 73% for ICSI) as was the viable pregnancy rate (51.4% for IVF and 55% for ICSI). No cycles failed to form blastocysts. CONCLUSIONS: The progression to blastocyst and the likelihood of conceiving a viable pregnancy were unaltered by ICSI. Thus it seems appropriate for programs to offer BT to patients undergoing ICSI using the same inclusion criteria applied to their IVF patients. PMID- 12735387 TI - Physiopathological aspects of corpus luteum defect in infertile patients with mild/minimal endometriosis. AB - PURPOSE: We describe a physiopathological model to the luteal insufficiency of infertile patients with mild/minimal endometriosis with normal hormone measurements in the early follicular phase. METHODS: We designed a case-control study with 24 patients, 14 fertile with in-phase endometrium (control group) and 10 infertile with mild/minimal endometriosis and luteal insufficiency (study group). The histologic dating of endometrium was performed during cycle days 23 25 and serum TSH, FSH, LH, prolactin, and estradiol levels were measured during the early follicular phase (cycle day 3). Progesterone serum levels were measured in three different occasions during the luteal phase. RESULTS: Patients with out of-phase endometrium have lower estradiol levels (P = 0.031) and decreased progesterone secretion (P = 0.012) during the late luteal phase. Serum prolactin, TSH, FSH, and LH levels were similar between the groups (P > 0.05). CONCLUSIONS: The physiopathology of luteal phase defect in infertile patients with mild/minimal endometriosis is associated with a small and large luteal cells dysfunction, characterized by abnormal follicular phase (lower estradiol serum levels) and lower progesterone LH-dependent secretion. PMID- 12735390 TI - Self-reported drinking-game participation of incoming college students. AB - Drinking games are associated with excess alcohol use and alcohol-related problems, yet it is unclear whether they are unique to the college environment or whether students come to college familiar with such games. The authors queried 1,252 students attending voluntary summer orientation programs about their experiences with drinking games. A majority (63%) indicated they had played drinking games and viewed them as a means to get drunk quickly and to socialize, control others, or get someone else drunk. Logistic regression analyses revealed that familiarity with drinking situations was associated with a greater likelihood of playing drinking games. Students who reported drinking more frequently and consuming greater quantities of alcohol than others, having lifetime marijuana use, and initiating alcohol consumption between the ages of 14 and 16 years were significantly more likely to have participated in drinking games. These findings demonstrate the importance of considering students' participation in drinking games when campus officials address alcohol use. PMID- 12735391 TI - Health behaviors, self-rated health, and quality of life: a study among first year Swedish university students. AB - The authors conducted a baseline investigation of male and female university students' health behaviors and self-rated health and quality of life (QOL). The study population consisted of all full-time, first-year students registered in a comprehensive study program offered at a Swedish university in autumn 1998. In spring 1999, the researchers sent self-administered questionnaires dealing with health status, lifestyle, and living conditions to the students at their home addresses. Male respondents used tobacco, were frequent drinkers, and engaged in binge drinking in larger proportions than expected by chance. A majority of the respondents rated their physical and psychological health as very good or good, but male students' ratings were higher than those of female students, whereas the males' average scores on self-perceived QOL were lower than those of females. Both male and female students' self-perceived QOL was more strongly associated with self-rated psychological than with physical health. PMID- 12735392 TI - Increased risk of alcohol abuse among college students living on the US-Mexico border: implications for prevention. AB - Alcohol abuse is a serious problem for students on college campuses. The authors conducted a cross-sectional study to (1) compare measures of alcohol-risk behaviors among college students attending a university on the US-Mexico border with other students and (2) identify factors associated with risk behaviors among border students. They used a self-administered survey to collect data from 286 freshman and sophomore students. Rates of binge drinking were higher among study respondents than among respondents in a national survey of freshman and sophomore college students (46% vs 42% to 45%, respectively). Drinking in high school and drinking in Mexico were significantly associated with all outcomes (binge drinking, drinking and driving, and riding with a drinker). The findings underscore the need for targeted prevention programs among this high-risk population. PMID- 12735393 TI - A note on logistic regression and odds ratios. PMID- 12735394 TI - Reaction to the September viewpoint on meningococcal vaccine. PMID- 12735395 TI - Introduction to special section on three-generation studies. AB - Interest in lifespan research and cross-generational associations in parenting practices and child behaviors has grown rapidly in recent years. The four papers presented in this journal test three key intergenerational research questions regarding intergenerational continuities for externalizing behaviors, using different 3-generational samples. PMID- 12735396 TI - Continuity of parenting practices across generations in an at-risk sample: a prospective comparison of direct and mediated associations. AB - A prospective model of parenting and externalizing behavior spanning 3 generations (G1, G2, and G3) was examined for young men from an at-risk sample of young adult men (G2) who were in approximately the youngest one third of their cohort to become fathers. It was first predicted that the young men in G2 who had children the earliest would show high levels of antisocial behavior. Second, it was predicted that G1 poor parenting practices would show both a direct association with the G2 son's subsequent parenting and a mediated effect via his development of antisocial and delinquent behavior by adolescence. The young fathers had more arrests and were less likely to have graduated from high school than the other young men in the sample. Findings were most consistent with the interpretation that there was some direct effect of parenting from G1 to G2 and some mediated effect via antisocial behavior in G2. PMID- 12735397 TI - Angry and aggressive behavior across three generations: a prospective, longitudinal study of parents and children. AB - This investigation examined intergenerational continuities in both angry, aggressive parenting and also the angry, aggressive behavior of children and adolescents. Data from 75 G2 youth (26 men, 49 women, M = 22-years old), their mothers (G1), and their G3 children (47 boys, 28 girls, M = 2.4-years old) were included in the analyses. The prospective, longitudinal design of the study, which included observational and multiinformant measures, overcame many of the methodological limitations found in much of the earlier research on intergenerational transmission. The results demonstrated a direct connection between observed G1 aggressive parenting and observed G2 aggressive parenting from 5 to 7 years later. G2 aggressive behavior as an adolescent and G3 aggressive behavior as a child were related to parenting behavior but not directly to one another. The results were consistent with a social learning perspective on intergenerational continuities in angry and aggressive behaviors. PMID- 12735398 TI - Cross-generational transmission of aggressive parent behavior: a prospective, mediational examination. AB - The intergenerational transmission of aggressive parenting behavior was examined within the context of a prospective longitudinal study of adolescent and young adult adjustment. Thirty-nine young adults (G2; 33 females, 6 males) who had participated in early phases of this study with their parents (G1) continued their involvement with their young children (G3; 17 females, 22 males, mean age = 2.6) several years later. Data included direct observation of parent-adolescent (G1-G2) and parent-child (G2-G3) interactions as well as self-reports. Analyses demonstrated directly observed cross-generational continuity in aggressive parenting from G1 to G2 some 6-7 years later. However, the results also showed that adolescent aggressive behavior served as the mediational link reducing the direct path from G1 to G2 aggressive parenting to nonsignificant levels. The results are consistent with a social interactional model of intergenerational continuity of parenting behavior. PMID- 12735399 TI - Linked lives: the intergenerational transmission of antisocial behavior. AB - There is a strong assumption of intergenerational continuity in behavior patterns, including antisocial behavior. Using a 3-generation, prospective study design, we examine the level of behavioral continuity between Generation 2 (G2) and Generation 3 (G3), and the role of economic disadvantage and parenting behaviors as mediating links. We estimate separate models for G2 fathers and G2 mothers. Data are drawn from the Rochester Youth Development Study, a longitudinal study begun in 1988 during G2's early adolescence (n = 1,000), which has collected prospective data on G2, their parents (G1), and now their G3 children. Results show that intergenerational continuity in antisocial behavior is evident, albeit somewhat modest. Parenting styles and financial stress do play a mediating role, although their effects vary by G2's gender. In general, adolescent delinquency plays a larger role in linking the generations for G2 fathers, whereas parenting behaviors and financial stress play a larger role for G2 mothers. PMID- 12735400 TI - Theoretical and methodological considerations in cross-generational research on parenting and child aggressive behavior. AB - The four studies in this special issue represent important advances in research on the intergenerational transmission of aggressive behavior. In this commentary, we review the key features and findings of these studies, as well as our own cross-generational study of aggression, the Columbia County Longitudinal Study. Next, we consider important theoretical issues (e.g., defining and operationalizing "aggression" and "parenting"; assessing reciprocal effects of parenting and child aggression; identifying the ages at which aggression should be assessed across generations; broadening the investigation of contextual and individual factors). We then discuss several methodological issues (e.g., determining the most informative measurement intervals for assessing prospective effects; sampling considerations; measuring potential moderating and mediating variables that might explain cross-generational continuities and discontinuities in parenting and aggression). Finally, we raise implications of cross generational research for designing interventions targeting the reduction and prevention of child aggression. PMID- 12735401 TI - Advancing our understanding of intergenerational continuity in antisocial behavior. AB - This commentary reviews the major findings of this set of 4 papers on intergenerational continuity in antisocial behavior; it identifies strengths and remaining challenges, and discusses potential policy implications of the research. As a group, these researchers have raised the methodological bar for future work in this area, using prospective designs with multiple informants and methods to test the influences of G2 parenting and adolescent antisocial behavior in mediating continuity between G1 parenting and G3 early disruptive behavior. The pattern of findings is discussed with respect to gender of G2 and social context. The inherent challenges of conducting intergenerational research are also highlighted, within the context of offering recommendations for improving future intergenerational investigations and their feasibility. PMID- 12735403 TI - Control beliefs as a mediator of the relation between stress and depressive symptoms among inner-city adolescents. AB - Adolescents' control beliefs were examined as a mediator of the relation between stress and depressive symptoms among a diverse sample of 445 inner-city adolescents. Results indicated that control beliefs significantly mediate the relation between stress and depressive symptoms. The specific direct effects of six individual stress domains (peer, family, school, neighborhood, economic, discrimination) on control beliefs and depressive symptoms were also examined. Results showed that (1) economic stress relates to adolescents' control beliefs, (2) family stress relates to adolescent depressive symptoms, and (3) peer stress relates to both control beliefs and depressive symptoms. Secondary analyses revealed that control beliefs significantly mediate the specific relations between peer stress and depressive symptoms. Results were not found to vary across ethnic groups. The implications of these findings for adolescent mental health and preventive interventions targeting depression are discussed. PMID- 12735405 TI - Good medicine. PMID- 12735404 TI - Reading problems and depressed mood. AB - Although reading difficulties show well-established overlaps with disruptive behavior disorders in childhood, much less is known about reading-disabled children's vulnerability to emotional difficulties. Using longitudinal data from 6 assessments of boys in the Pittsburgh Youth Study, we found robust links between severe, persistent reading problems and increased risk for depressed mood in a community sample of boys aged 7 and 10 years at initial assessment, though not in those who had already entered their teens. These associations could not be accounted for in terms of selected family risks or comorbid disruptive behaviors; instead, the pattern of the findings pointed to the existence of more direct causal processes whereby reading problems influence younger boys' risk of depressed mood. PMID- 12735406 TI - Assessment of oxygenation in the fetus and newborn. PMID- 12735407 TI - Cost analysis of Down syndrome screening in advanced maternal age. AB - OBJECTIVE: To analyze the potential cost and efficacy of Down syndrome screening in the population with advanced maternal age. METHODS: Three screening methods defining Down syndrome risk for women with advanced maternal age were analyzed: advanced maternal age; advanced maternal age and maternal serum triple screen; and advanced maternal age, maternal serum triple screen and genetic sonogram. Costs for all tests and procedures were estimated. Procedure-related loss for amniocentesis was assumed to be 1:200. Efficacy was defined as: number of amniocenteses performed, number of Down syndrome cases detected, procedure related losses, Down syndrome cases detected per fetal loss, cost per Down syndrome case detected and total cost of screening. RESULTS: In 1999 in the USA, there were 530,610 women with advanced maternal age at 16 weeks' gestation carrying an estimated 4,043 fetuses with Down syndrome. Screening by maternal age alone would result in the 100% detection of Down syndrome cases, but would require over 530,000 amniocenteses and result in 2,653 procedure-related losses. Combining age with serum screen and genetic sonogram would detect 97.6% of Down syndrome cases, but would require only 119,791 amniocenteses and result in 599 procedure-related losses. The projected cost per Down syndrome case detected using age screening is 219,109 dollars versus 155,992 dollars using serum screen and genetic sonogram. CONCLUSIONS: The combination of advanced maternal age, maternal serum screen and genetic sonogram would result in the fewest procedure related losses and lowest cost per Down syndrome case detected. PMID- 12735408 TI - Prevalence of prenatal drinking assessed at an urban public hospital and a suburban private hospital. AB - BACKGROUND: Although the prevalence of fetal alcohol syndrome (FAS) varies within the population, few data are available concerning variation in the prevalence of prenatal drinking. METHODS: Postpartum women delivering singleton infants at two Atlanta hospitals in 1993 or 1994 were interviewed. Those delivering infants who were small for gestational age (SGA) (n = 638) were over-sampled relative to those delivering infants with birth weights that were appropriate for gestational age (AGA) (n = 247). The prevalence of prenatal drinking was estimated as a weighted average of reports from mothers of SGA and AGA infants. Estimates of the prevalence of FAS come from the Metropolitan Atlanta Congenital Defects Program (MACDP) of the Centers for Disease Control and Prevention. RESULTS: The prevalence of first-trimester drinking was half that reported for the three previous months (private hospital: 72% vs. 35%; public hospital: 52% vs. 28%). Most women (85%) reported abstaining throughout the second trimester. Fewer than 10% of women delivering at the public hospital (7.5%), but one-quarter of those delivering at the private hospital, reported third-trimester drinking. Binge, moderate and heavy drinking in pregnancy were more common among women delivering at the public hospital. Eight infants born at the public hospital during this period, but none of those born at the private hospital, were identified as possibly having FAS; four of the eight were identified as probably having FAS. CONCLUSIONS: These results have implications for health education programs. For example, obstetricians in private practice may wish to reaffirm their advice to abstain from drinking in the third trimester. They also suggest that prenatal abstinence programs be targeted at populations identified as most likely to engage in risky drinking. PMID- 12735409 TI - Do antenatal classes benefit the mother and her baby? AB - OBJECTIVE: To examine the characteristics of women attending antenatal classes and evaluate the effects of these classes on mothers' and babies' health. METHODS: A population-based observational study on care during pregnancy, delivery and in the postnatal period was carried out in 1995-96. A total of 9004 women resident in 13 regions of Italy who delivered in a 4-month period were interviewed. The outcomes studied were attendance at antenatal classes, Cesarean section, bottle feeding, satisfaction with the experience of childbirth, knowledge of contraception, breast feeding and baby care. RESULTS: A total of 2065 (23.0%) women attended antenatal classes. Women without previous children, those with a higher level of education and office workers were more likely to attend classes. Women who attended antenatal classes had a much lower risk of Cesarean section and were about half as likely to bottle feed while in hospital compared with non-attenders. They received better information on contraception, breast feeding and baby care. Women who attended classes and applied the techniques learned were more satisfied with the experience of childbirth. CONCLUSION: Antenatal classes seem to improve women's knowledge and competence. This may provide a defence against the tendency to overmedicalize pregnancy and childbirth. PMID- 12735411 TI - Evaluation of non-reassuring fetal heart rate patterns with fetal pulse oximetry combined with vibratory acoustic stimulation. AB - OBJECTIVE: To determine the value of fetal pulse oximetry and vibratory acoustic stimulation in the presence of non-reassuring fetal heart rate patterns during labor. DESIGN: Prospective study in women monitored by cardiotocography and fetal pulse oximetry during labor. MATERIALS AND METHODS: During a period of 18 months, 907 consecutive parturients in labor were monitored by cardiotocography. Out of these women, 63 were selected on the basis of a non-reassuring fetal heart rate tracing during the first stage of labor. In these cases, fetal pulse oximetry was applied. Vibratory acoustic stimulation was applied in fetuses without spontaneous reactivity in order to evaluate the fetal status. RESULTS: Our cases were classified into three groups, according to the lower fetal oxygen saturation levels, from the time of oximetry application until delivery. Group A consisted of 29 cases where fetal oxygen saturation levels were > or = 41%, group B (20 cases) with fetal oxygen saturation of 31-40% and group C (14 cases) with levels of < 30%. Spontaneous reactivity was observed in 15 fetuses of group A and seven of group B, while no case of reactivity was noted in group C. Vibratory acoustic induced reactivity was associated with low fetal oxygen saturation levels. The mean umbilical artery pH levels were 7.29 +/- 0.051 in group A, 7.21 +/- 0.057 in group B and 7.04 +/- 0.05 in group C. CONCLUSION: Fetal pulse oximetry should be indicated not only in fetuses without any reactivity but also in those with induced reactivity, after the application of vibratory acoustic stimulation. PMID- 12735410 TI - The relationship between placental histopathology findings and perinatal outcome in preterm infants. AB - OBJECTIVE: To determine the correlation between placental histopathology findings and perinatal outcome in preterm infants. METHODS: Placental histopathology in 774 neonates delivered at 24-32 weeks between 1992 and 2000 was classified as follows: 254 (33%) had histological chorioamnionitis, 263 (34%) had coagulation related lesions, 228 (30%) had vasculopathy. Perinatal outcome was compared between cases positive and negative for each histopathological classification. RESULTS: Histological chorioamnionitis occurred in 46% of cases with premature rupture of membranes and 45% with preterm labor. Positivity versus negativity for histological chorioamnionitis was associated with earlier presentation (191 vs. 205 days, p = 0.0001) and delivery (199 days vs. 209 days, p = 0.0001), increased risk of intraventricular hemorrhage (71% vs. 23%, p = 0.001, odds ratio (OR) 2.2), bronchopulmonary dysplasia (26% vs. 15%, p = 0.0001, OR 2), retinopathy (36% vs. 24%, p = 0.001, OR 1.8), neonatal sepsis (28% vs. 13%, p = 0.0001, OR 2.5) and neonatal death (12% vs. 7%, p = 0.012, OR 2). Vasculopathy versus no vasculopathy was associated with decreased birth weight (1245 g vs. 1341 g, p = 0.011), decreased Apgar score at 5 min (20% vs. 13%, p = 0.011, OR 1.7) and necrotizing enterocolitis (6% vs. 2%, p = 0.001, OR 4). Cases positive for coagulation-related lesions correlated only with necrotizing enterocolitis (5% vs. 2%, p = 0.02, OR 2.6). CONCLUSIONS: The presence of histological chorioamnionitis significantly increases the risk of earlier delivery and neonatal mortality. Vascular and coagulation placental findings increase the risk of necrotizing enterocolitis. PMID- 12735413 TI - Can we prevent postnatal depression? A randomized controlled trial to assess the effect of continuity of midwifery care on rates of postnatal depression in high risk women. AB - OBJECTIVE: To evaluate the effectiveness of continuous midwifery care in reducing rates of postnatal depression in women with histories of depression. METHODS: Fifty-one women from a sample of 98 pregnant women with histories of major depressive disorder were randomly allocated, at antenatal booking, to continuous midwifery care. The remaining 47 women received standard maternity care. A total of 87 women (44 treatment, 43 control) completed baseline assessments (after randomization) and outcome assessments (at 3 months postpartum). RESULTS: Of the women allocated to continuous midwifery care, 88% complied in full with their allocated treatment protocol. Forty-nine percent of women had an episode of illness in pregnancy (DSM-III-R case of major or minor depression), 26% developed a new episode of illness after antenatal booking, and 23% were depressed in the first 3 months postpartum. There were no differences between treatment conditions in total rates, or rates within the context of social adversity, of antenatal depression, depressions which occurred post-booking, postnatal depression, and the duration of depressive episodes. CONCLUSIONS: While continuous midwifery care had no impact on psychiatric outcome, it was highly successful at engaging women in treatment and therefore has an important contribution to make in the care of child-bearing women with mental health problems. PMID- 12735412 TI - Amniotic levels of nitric oxide in women with fetal intrauterine growth restriction. AB - OBJECTIVE: Many biochemical observations have shown that nitric oxide (NO) is involved in the vascular angiogenic activity of the fetoplacental unit. The aim of this study was to determine whether NO is implicated in the pathogenesis of intrauterine growth restriction (IUGR). METHODS: We retrospectively assessed amniotic fluid NO from second-trimester amniocentesis of 20 healthy normotensive women who subsequently developed IUGR and 20 controls. The same women were re assessed at the third trimester when IUGR had developed and when the same 20 controls had shown normal pregnancy. Amniotic fluid NO was detected by discontinuous spectrophotometry and the Griess reaction. RESULTS: At the second trimester, NO levels in women with subsequent IUGR were significantly lower than in controls (4.1 +/- 0.2 microg/mg creatinine vs. 6.02 +/- 1.57 microg/mg creatinine, p < 0.001). At the third trimester, in women with IUGR, NO levels were significantly higher than in normal pregnancies (7.4 +/- 1.5 vs. 5.02 +/- 0.9 microg/mg creatinine, p < 0.001), and directly correlated with gestational age when growth restriction was diagnosed (r = 0.69, p < 0.001). CONCLUSIONS: Low levels of NO during the early second trimester may represent an impaired stimulus to vascular formation and endothelial regulation, inducing placental disease and subsequent fetal growth restriction. High levels of amniotic fluid NO during the third trimester may represent a compensation factor for maintaining adequate uteroplacental perfusion in pregnancies with IUGR. PMID- 12735415 TI - Uterine rupture associated with castor oil ingestion. AB - A woman at 39 weeks' gestation with a previous Cesarean delivery had severe abdominal pains and rupture of membranes shortly after ingesting 5 ml of castor oil. Forty-five minutes later, repetitive variable decelerations prompted a Cesarean delivery. At surgery, a portion of the umbilical cord was protruding from a 2-cm rupture of the lower transverse scar. PMID- 12735414 TI - Large fetomaternal hemorrhage: clinical presentation and outcome. AB - AIM: To evaluate the incidence and outcome of all neonates with demonstrated fetomaternal hemorrhages > or = 20 ml and to assess possible predictors of large fetomaternal hemorrhage and outcome. METHODS: Retrospective data analysis 1987 2000. Clinical data included antenatal events, method of delivery, condition at birth, hematology results, treatment and outcome. RESULTS: Sixteen infants were identified and treated for fetomaternal hemorrhage. Adverse outcome occurred in five infants (31%). Outcome was predicted by postnatal presentation and initial hemoglobin. CONCLUSION: Adverse outcome amongst neonates with large fetomaternal hemorrhage is high. Outcome is better predicted by initial hemoglobin than volume of hemorrhage as per the Kleihauer test. PMID- 12735416 TI - The Zavanelli maneuver in two cases of shoulder dystocia. AB - Several reports describe the replacement of the partially delivered fetus into the vagina followed by Cesarean section after failed vaginal delivery. Undeliverable shoulder dystocia is by far the most common cause of failed vaginal delivery. The objective of this report was to review the safety and to reconsider when to perform the Zavanelli maneuver in the management of shoulder dystocia. We report two cases of severe shoulder dystocia in which commonly recommended therapeutic options failed. Cephalic replacement was performed with ease followed by Cesarean section. Endometritis was the maternal complication in both cases. Both infants had transient Erb's palsy. Although traditionally the Zavanelli maneuver is a method of last resort in the management of shoulder dystocia, consideration should be given to its use earlier in the treatment of obstructed vaginal delivery. PMID- 12735417 TI - Persisting challenges in plasma endocrinology: reference values and endocrine tests. AB - The analysis of plasma hormone concentrations is of fundamental importance for the diagnosis and treatment of endocrine diseases. Although hormone analyses are performed in huge numbers in all hospitals on a daily basis, the interpretation of the resulting plasma hormone concentrations can be difficult. In addition to the effects of the underlying disease, biological and analytical issues affect hormone concentrations. Therefore, adequate reference values and strict standardisation of sampling and analytical procedures are very important for the final interpretation of the results of hormone analysis. PMID- 12735418 TI - Cardiovascular disease in sub-Saharan Africa: a disaster waiting to happen. AB - The looming threat posed by the emergence of cardiovascular disease (CVD) in sub Saharan Africa (sSA) is underestimated and often denied. The health services and societies struggle to cope with the direct effects of poverty, war, fragile social and economic structures and AIDS. The threat of CVD seems less direct and few reliable data are available. This has resulted in neglecting serious warning signs on the emergence of CVD in sSA. This short review deals with the strong increase in a number of risk factors for CVD in certain areas, necessitating preventive measures to lighten the 'double burden of disease' in this part of the world. PMID- 12735419 TI - Establishment of reference values for endocrine tests. III: Primary aldosteronism. AB - BACKGROUND: In our laboratory well-defined reference values for the screening test and confirmation test used in the diagnosis of primary aldosteronism were lacking. In this study we established the reference-values of the plasma aldosterone concentration (PA), plasma renin activity (PRA) and PA/PRA ratio after a two-hour upright period, and of the urinary aldosterone excretion after oral sodium loading. METHODS: Fifty healthy volunteers, equally distributed according to sex and aged between 20 and 70 years, went through the screening and confirmation test of primary aldosteronism. PA, PRA and the PA/PRA ratios were measured after a two-hour upright period (screening test). Urinary aldosterone excretion was determined in two 24-hour urine samples after an oral suppletion of 6 g NaCl a day for five days (confirmation test). RESULTS: The following reference values were established: PA (after two-hour upright position) <0.03 1.05 nmol/l (mean: 0.47), PA/PRA ratio 0.05-0.47 (mean: 0.15) and urinary aldosterone excretion after sodium loading <3.0-47.0 nmol/24h (mean: 10.5). PRA showed a significant decrease with advancing age: median values in the 3rd to 7th decade are 3.9, 3.5, 2.5, 1.6 and 2.1 ng A1/ml/h respectively (p=0.04). PA was lower in subjects > or = 50 years old. Age did not affect the PA/PRA ratio or the urinary aldosterone excretion. There were no significant differences between the sexes in any of the above-mentioned parameters. CONCLUSION: In this study we established reference values for the screening and confirmation test used in the diagnosis of primary aldosteronism. PMID- 12735420 TI - Evaluation of endocrine tests. A: the TRH test in patients with hyperprolactinaemia. AB - BACKGROUND: In a previous study, we determined reference values for basal and thyrotropin-releasing hormone (TRH)-stimulated plasma concentrations of prolactin (PRL). The aim of the present study was to determine the clinical usefulness of the PRL response to TRH in the work-up of patients with hyperprolactinaemia. METHODS: We studied 92 consecutive patients referred for evaluation of hyperprolactinaemia. Patients with confirmed hyperprolactinaemia were divided into three groups: group A (pharmacological hyperprolactinaemia; n=2), group B (pathological hyperprolactinaemia; n=6) and group C (all other patients). Patients in group C underwent MRI of the pituitary and were subdivided into C1 (normal pituitary on MRI; n=6), C2 (slightly abnormal MRI; n=21), and C3 (evident microadenoma or macroadenoma on MRI; n=25 and 12, respectively). The MRI was technically insufficient in four patients. Basal PRL as determined by fluoroimmunometric assay and the PRL response to 400 microg TRH were determined in all patients. RESULTS: Hyperprolactinaemia was confirmed in 83% of the referred patients. Non-response, defined as a <2.5-fold PRL increase after TRH, occurred in one patient (50%) in group A, in 66% of patients in group B and in 99% of patients in group C. Within group C, basal PRL was not different between group C1 and C2, but higher (p=0.06) in group C3. The absolute PRL increase after TRH did not differ between the three subgroups. The relative PRL increase was smaller (p=0.03) in group C3 but overlapped considerably with groups C1 and C2. All patients except one in group C were so-called non-responders. Basal PRL and absolute PRL increases after TRH correlated with the adenoma diameter on MRI (r=0.66, p=0.0002 and r=0.49, p=0.008, respectively). CONCLUSION: In patients referred for elevated serum PRL, hyperprolactinaemia should be confirmed under standardised conditions. The absolute or relative PRL increase after 400 microg TRH does not help to differentiate between patients with prolactinoma or idiopathic hyperprolactinaemia. Therefore, the TRH stimulation test is not useful in the work-up of hyperprolactinaemia. PMID- 12735421 TI - Photo quiz. Bilateral adrenal tumour. PMID- 12735422 TI - Bleomycin and scuba diving: to dive or not to dive? AB - Bleomycin is to treat patients with testicular cancer and lymphoma. Bleomycin can bind to DNA and chelate iron. The resulting complex can form an intermediate capable of interacting with oxygen to produce reactive oxygen species, particularly superoxide. Administrating high-inspired oxygen concentrations (e.g. during anaesthesia or acute illness) has been reported to exacerbate pulmonary injury. The duration of risk after bleomycin chemotherapy is unknown. Here we discuss our advice to a young male patient, who was successfully treated with bleomycin for testicular cancer, concerning the safety to return to scuba diving. Since scuba divers are exposed to high partial oxygen pressures (depending on the depth of the dive) we discouraged this patient from resuming scuba diving. PMID- 12735423 TI - Tension pneumopericardium caused by positive pressure ventilation complicating anaerobic pneumonia. AB - A 22-year-old man was admitted with pneumonia. He was immediately intubated and positive pressure ventilation was initiated. Blood and sputum cultures showed Bacteroides fragilis and Corynebacterium sp., which were treated with metronidazole and clindamycin. Three weeks later his blood pressure suddenly dropped with an elevation of the central venous pressure. Chest X-ray revealed a pneumopericardium. A parasternal mediastinotomy with partial pericardiectomy was immediately performed. On opening the pericardium his blood pressure normalised. The patient gradually recovered and six weeks after admission he was extubated. Two weeks later he was discharged. A pneumopericardium without previous thorax trauma is very rare and early recognition is imperative because a tension pneumopericardium with cardiac tamponade may develop, as happened in this case. A tension pneumopericardium has to be treated with immediate pericardiocentesis followed by partial pericardiectomy to avoid recurrence. PMID- 12735424 TI - Cardiac failure following group A streptococcal infection with echocardiographically proven pericarditis, still insufficient arguments for acute rheumatic fever: a case report and literature update. AB - We recently encountered a 49-year-old female who developed fever due to group A streptococcal (GAS) bacteriaemia spreading to an abscess in the iliac muscle and a bacterial monarthritis of the right knee with a sterile arthritis of her left knee. Treatment was started with a six-week course of intravenous penicillin. She developed a mitral valve insufficiency and pericarditis on the tenth day of admission. In the third week heart failure developed with, on echocardiograph, a high output left ventricular failure without signs of valvulitis or myocarditis. Using a diuretic regimen she was recompensated. Because of the pericarditis with mitral valve insufficiency corticosteroids were given, which had a rapid beneficial effect. A discussion follows on the position of acute rheumatic fever versus post-streptococcal reactive arthritis in this clinical picture and the literature is updated. PMID- 12735425 TI - A study on the rehabilitation of cognitive function and short-term memory in patients with Alzheimer's disease using transcutaneous electrical nerve stimulation. AB - In some previous studies, transcutaneous electrical nerve stimulation (TENS) to the back was shown to improve non-verbal short-term and long-term memory as well as verbal fluency in patients in the early stage of Alzheimer's disease (AD). Moreover, the patients' physical, social and affective functions were also improved. In this study, the effects of TENS applied to the face were studied in patients with mild and severe AD. Fourteen patients with probable AD were used as subjects. To evaluate the effects of treatment, the subjects underwent two kinds of neuropsychological tests as well as a pupillary light reflex test. The pupillary light reflex test was chosen because it has been proven to show poorer results in cases of senile dementia. These three tests were carried out before the treatment, immediately after the treatment and 6 months after the treatment. Improvements were seen in subjects in the TENS groups and the effects of treatment were more significant in mild AD patients than in severe AD patients. However, the improvements were not maintained 6 months after the treatment. The possible mechanisms underlying the beneficial effects of TENS in AD patients are discussed in this paper. PMID- 12735426 TI - Influence of cool-down exercise on autonomic control of heart rate during recovery from dynamic exercise. AB - The recovery of post-exercise heart rate (HR) is enhanced by a procedure of cooling down; however, the mechanism of this facilitated reduction is unknown. To determine whether more cardiac vagal reactivation is associated with a decrease in HR following cool-down exercise, we examined high-frequency R-R interval variability (HF, 0.15-0.40 Hz), an index of cardiac vagal tone, in six young healthy male subjects with a comparison between sitting rest after 6 minutes of cool-down cycling at 20% peak oxygen uptake (VO2peak) and sitting complete rest, following 5 min of upright cycle exercise at 70% VO2peak. During the last minute of exercise, there was no difference in HR between the two exercise tests before performing or not performing cool-down exercise (mean +/- SE, 148.7 +/- 6.9 versus 149.7 +/- 7.0 beats/min, respectively, by a Wilcoxon signed-ranks test). After exercise, a similar initial rapid decrease in HR and subsequent decline was observed during the first 6 min of recovery, regardless of cool-down exercise. However, the resting HR average following cool-down exercise was significantly lower than the corresponding HR without cool-down exercise (92.1 +/- 3.0 versus 100.8 +/- 3.6 beats/min, P < 0.05). The corresponding HF amplitude was slightly, but significantly greater with than without cool-down exercise (10.6 +/- 2.2 versus 9.0 +/- 1.9, P < 0.05). Thus the increase in HF amplitude corresponded to a decrease in resting HR following cool-down exercise, as compared to complete rest without cool-down exercise. Therefore, we conclude that after moderate exercise, the decrease in resting HR following cool-down exercise is associated with an increase in cardiac vagal tone. PMID- 12735427 TI - Adaptive EEG spike detection: determination of threshold values based on conditional probability. AB - Determination of the threshold value for automatic EEG spike detection was investigated adopting conditional probability. An adaptive spike detection method was constructed and evaluated. A discriminant function for detecting spikes was obtained by conditional probability calculated from the EEG spike data. The relationship among false-negatives, false-positives and threshold values for the discriminant function was investigated. An adaptive detection algorithm was developed by combining different threshold values. False-negative and false positive rates for spike detection depended on the threshold values. The adaptive spike detection algorithm achieved a high detection rate and accuracy. The advantage of the proposed method is to construct an adaptive detection algorithm by combining the threshold values according to the purpose of spike detection. Since the threshold can be easily changed in the proposed method, it is practically effective for clinical use. PMID- 12735428 TI - Roles of reactive oxygen species in monocyte activation induced by photochemical reactions during photodynamic therapy. AB - This study attempts to investigate the mechanism of the vascular shut down (VSD) effect during photodynamic therapy (PDT) with zinc coproporphyrin III tetrasodium salt as a photosensitizer. PDT is a treatment based on photochemical reactions and the resultant cytotoxic reactive oxygen species (ROS). Platelet thrombus formation leading to stasis observed in vivo during PDT is called the VSD effect. Leukocytes play an important role in the VSD effect in vivo, but the mechanism how activated monocytes generate ROS is not known in detail. To evaluate ROS generation by activated monocytes is especially important to clarify leukocyte endothelium interactions in the VSD mechanism. The dichlorofluorescein fluorescence intensity of monocytes with four types of free radical scavenger was investigated by confocal laser scanning microscopy. The fluorescence intensity of monocytes that had been incubated with superoxide dismutase and incubated and added with L-histidine was decreased by about 20 and 30%, respectively. The result affirms the predominant role of singlet oxygen and superoxide anion radicals in monocyte activation in the VSD effect during PDT. PMID- 12735429 TI - Development of a portable bruxism monitoring and analysis device equipped with a microcomputer and its practical application. AB - The purposes of this study were to develop and verify the a portable nocturnal bruxism monitoring and analysis device equipped with a microcomputer, and to clinically apply the device to know the actual conditions of bruxism patients. EEPROM was installed in the device for the data recording, and after the data collection, the recorded data was entered into a personal computer via serial port. After confirming the accuracy of the device, a total of 30 subjects were enrolled in this study to monitor their bruxism activities for 3 nights. Bruxism self-aware group consisted of 14 subjects, 7 males and 7 females, and unaware group consisted of 16 patients, 8 males and 8 females. Most of the subjects reported that the new device was easy to handle. The average bruxism time per hour and the average bruxism lasting time were 223.8 +/- 112.0 and 3.9 +/- 2.9 s in the self-aware group, and 49.3 +/- 38.3 and 0.8 +/- 0.7 s in the unaware group, respectively. The bruxism self-aware group showed statistically longer average bruxism time per hour and the average bruxism lasting time. It was confirmed that the new bruxism monitoring and analysis device is practical for clinical application to monitor and analyze the electromyographic activities. PMID- 12735430 TI - A comparison of abdominal ECG and Doppler ultrasound for fetal heart rate detection. AB - A fetal monitor has been developed for the measurement of the fetal and maternal heart rates from maternal abdominal electrocardiogram during pregnancy and labor for ambulatory monitoring. Developed algorithm of the fetal monitor is based on digital filtering, adaptive thresholding. statistical properties in the time domain and differencing of local maxima and minima. Five volunteers with low risk pregnancies, between 35 to 40 weeks of gestation and no evidence of labor, were studied for the fetal heart rate detection. A Doppler ultrasound fetal monitor was used to compare the accuracy of the measurement system. Results showed an average percent rms difference (PRD) of 5.32% in comparison with the reference ultrasound method. The fetal heart rates curve remained inside a +/- 5 beats/min limit relative to the reference ultrasound method for 84.1% of the time. PMID- 12735431 TI - Development of an EMG recording device from stimulation electrodes for functional electrical stimulation. AB - A device that records voluntary EMG from a pair of surface stimulation electrodes has been developed for functional electrical stimulation (FES). The device can apply to a small muscle on which it is difficult to locate both the stimulation electrodes and recording electrodes. The device utilizes photo-MOS relays to disconnect the stimulator when it is not active, to ground the electrodes after delivering the stimulation pulses, and to drop the gain of the EMG amplifier during the stimulus period. The device is able to detect the voluntary EMG of a small muscle from the stimulation electrodes for the EMG-controlled FES system. PMID- 12735432 TI - The high zero-flow pressure phenomenon in coronary circulation: a simulation study. AB - A dynamic simulation was performed in order to study the high zero-flow pressure phenomenon by using an electronic model of the coronary vessel. The objective of this study was to establish a relationship between the diastolic arterial pressure and diastolic arterial flow. The simulated study showed that this relationship is approximately linear with a zero flow pressure intercept of approximately 40 mmHg. This result is consistent with the results obtained in animal experimentation conducted by others. PMID- 12735433 TI - Closed-loop stripping analysis of synthetic musk compounds from fish tissues with measurement by gas chromatography-mass spectrometry with selected-ion monitoring. AB - Synthetic musk compounds have been found in surface water, fish tissues, and human breast milk. Current techniques for separating these compounds from fish tissues require tedious sample clean-up procedures. A simple method for the determination of synthetic musk compounds in fish tissues has been developed. Closed-loop stripping of saponified fish tissues in a 1-1 Wheaton purge-and-trap vessel is used to strip compounds with high vapor pressures such as synthetic musks from the matrix onto a solid sorbent (Abselut Nexus). This technique is useful for screening biological tissues that contain lipids for musk compounds. Analytes are desorbed from the sorbent trap sequentially with polar and nonpolar solvents, concentrated, and directly analyzed by high resolution gas chromatography coupled to a mass spectrometer operating in the selected ion monitoring mode. In this paper, we analyzed two homogenized samples of whole fish tissues with spiked synthetic musk compounds using closed-loop stripping analysis and pressurized liquid extraction (PLE). The analytes were not recovered quantitatively but the extraction yield was sufficiently reproducible for at least semi-quantitative purposes (screening). The method was less expensive to implement and required significantly less sample preparation than the PLE technique. PMID- 12735434 TI - Enantioselective liquid chromatography of C3-chiral 2,3-dihydro-1,2,5 benzothiadiazepin-4(5H)-one and thione 1,1-dioxides on polyacrylamide- and polysaccharide-based chiral stationary phases. AB - Optically active synthetic and semisynthetic polymers were utilized as chiral stationary phases (CSPs) for the direct chromatographic enantioseparation of a series of 8-chloro-2,3-dihydro-3-methyl-1,2,5-benzothiadiazepin-4(5H)-one and thione 1,1-dioxide. Evaluation of stereochemical integrity of chiral analytes was assessed by enantioselective temperature and flow-dependent HPLC. A stopped-flow high-performance liquid chromatography (sfHPLC) procedure was developed for the determination of the rate constants and free energy barriers of enantiomerization of enantiomers of 8-chloro-2-(3-methylbut-2-enyl)-2,3-dihydro-3-methyl-1,2,5 benzothiadiazepin-4(5H)-thione 1,1-dioxide (compound 2) in the presence of Chiraspher and Chiralcel OD CSPs. In order to study the chiroptical properties of the individual enantiomers of analytes investigated, semipreparative chromatographic resolutions were performed. The assignment of the absolute configuration was empirically established by comparing the CD spectra of the separated enantiomers with those obtained from structural analogues. PMID- 12735435 TI - Simultaneous determination of neutral and acidic pharmaceuticals in wastewater by high-performance liquid chromatography-post-column photochemically induced fluorimetry. AB - An analytical method for the simultaneous determination of acidic and neutral pharmaceutical active compound (PhACs) residues in wastewater has been developed based on the combination of high-performance liquid chromatography (HPLC) and photochemically induced fluorimetry. The photoderivatization conditions for each particular PhAC have been assessed. Off-line optimization of the HPLC separation for both neutral and acidic compounds has been utilised and evaluated. Detection limits in the low ng/ml range have been achieved without sample pretreatment. By applying the developed analytical method combined with solid-phase extraction to real wastewater samples an enrichment factor of approximately two orders of magnitude can be obtained. PMID- 12735436 TI - Determination of zearalenone from wheat and corn by pressurized liquid extraction and liquid chromatography-electrospray mass spectrometry. AB - Zearalenone (ZON) was extracted from wheat and corn by using pressurized liquid extraction (PLE) and the PLE extracts were analyzed using liquid-chromatography mass spectrometry (LC-MS) without further clean-up procedures. A statistical design approach was applied to evaluate the influence of several extraction parameters such as temperature (40 degrees C; 80 degrees C; 120 degrees C), time (5 min; 10 min) and solvent extraction mixture [acetonitrile-water (9:1, v/v); methanol-water (8:2, v/v); methanol-acetonitrile (1:1, v/v)] on fortified cereals. The results showed a strong influence of the solvent composition on recovery of ZON. Quantification of the analytes was performed by LC-MS analysis of the raw extract using matrix-matched standard curves. The method performance was tested in the selected conditions (80 degrees C; 5 min; two cycles; methanol acetonitrile) on samples which had been previously used for an international proficiency test. Compared to the assigned value, the recovered ZON was 118% [relative standard deviation (RSD)=5.2%, n=3)] and 107% (RSD=2.2%, n=3) in wheat and corn, respectively. Therefore, PLE can be used for ZON extraction, achieving good performances and allowing for an automated handling of the sample extraction step. Successively, the influence of temperature and number of cycles was investigated on naturally contaminated corn. From these results it could be concluded that fortified experiments perfectly mimicked naturally contaminated samples. PMID- 12735437 TI - Theoretical and experimental study of recycle capillary gas chromatography with carrier gas propelled by a peristaltic pump. AB - A new technique of recycle capillary gas chromatography (RCGC) characterized by a very high separation efficiency of more than 10(6) theoretical plates has been developed to solve the problem of separation of isomers with similar physico chemical properties. The technique replaces the recycle valve by a peristaltic pump that propels the carrier gas. A general model has been developed for description of RCGC characteristics and experimentally verified on the retention behaviour of methane and the separation of a test pair of 3-methyl-1-butene and 2 methylbutane. PMID- 12735438 TI - Chromatographic properties of tetramethyl-p-silphenylene-dimethyl, diphenylsiloxane copolymers as stationary phases for gas-liquid chromatography. AB - Seven tetramethyl-p-silphenylene-dimethyl, diphenylsiloxane copolymers were coated on fused-silica capillary columns to evaluate their properties as stationary phases in gas-liquid chromatography. The capillary columns were tested concerning their selectivity, separation efficiency, column bleed, inertness, elution temperatures, and working range. The following characteristic properties of the silphenylene unit were found: (i) the impact of the silphenylene group on the chromatographic selectivity is similar to that of two dimethylsiloxy groups and half of a diphenylsiloxy group; (ii) silphenylene-siloxane copolymers offer reduced column bleed and increased maximum allowable operating temperature in comparison to polysiloxanes, since the backbone stiffing phenylene group enhances thermal stability; (iii) the elution temperatures of analytes are increased by 15 30 degrees C on silphenylene-siloxane copolymers compared to polysiloxanes; (iv) the silphenylene unit increases the glass transition temperature of the polymers resulting in elevated minimum allowable operating temperatures. PMID- 12735439 TI - Trace analysis of trichlorobenzenes in fish by microwave-assisted extraction and gas chromatography-electron-capture detection. AB - An analytical method consisting of extraction, clean-up, and analysis by gas chromatography-electron-capture detection (GC-ECD) was developed for the determination of trichlorobenzenes (TCBs) in fish samples. Two extraction methods, saponification and liquid-liquid extraction (S-LLE), and microwave assisted extraction (MAE), were evaluated. In both cases, n-pentane was used as the extraction solvent. For S-LLE, the recoveries ranged from 66.6+/-9.1% for 1 bromo-4-chlorobenzene (4-BCB) to 93.5+/-4.9% for 1,2,4-trichlorobenzene (1,2,4 TCB). The recoveries were significantly lower, between 31.0+/-3.9% for 1,2,3 trichlorobenzene (1,2,3-TCB) and 52.3+/-3.0% for 1,3,5-trichlorobenzene (1,3,5 TCB), in the absence of fish. Proteins and glycerides of the fish tissue seemed to compete with TCBs for the base, and hence decreased their decomposition rate. In the case of MAE, the recoveries were highly dependent on the pressure applied during extraction. At 5 bar, much higher recoveries were obtained, from 66.7+/ 15.6% for 4-BCB to 79.9+/-13.6% for 1,2,4-TCB, than at 1 bar. Sulfur formation was, however, observed at 5 bar, and interfered with the GC-ECD analysis of TCBs. Sulfur was adequately removed by copper powder treatment, which was shown not to affect the recovery of analytes. The recoveries of target analytes by S-LLE and MAE did not differ statistically (t-test, alpha = 0.01). Both methods were appropriate for the detection of TCBs at concentration levels typically observed in marine biota, i.e. approximately 1 ng/g. S-LLE was, however, more time consuming, and required larger volumes of high-purity organic solvents than MAE. PMID- 12735440 TI - Extraction and quantitative analysis of 1-aminocyclopropane-1-carboxylic acid in plant tissue by gas chromatography coupled to mass spectrometry. AB - We developed a new method for the determination of the ethylene precursor 1 aminocyclopropane-1-carboxylic acid (ACC) using quantitative GC-negative chemical ionisation MS as a detection and quantification system, in combination with isotope dilution using [2H4]ACC and an off-line solid-phase extraction. By derivatisation with pentafluorobenzyl bromide, ACC could easily be detected with m/z 280 being the most abundant ion. Determination of this component resulted in a detection limit of 10 fmol and a linear fit in the 100 fmol-100 pmol range. The combination of a rapid, high yield purification method with a stable derivatisation procedure and a sensitive detection method allowed the detection of ACC in samples as low as 100 mg fresh mass. PMID- 12735441 TI - Comprehensive chemical derivatization for gas chromatography-mass spectrometry based multi-targeted profiling of the major phytohormones. AB - In the present investigation we report selection of the N-methyl-N-(tert. butydimethylsilyl)trifluoroacetamide (MTBSTFA) reagent as the most comprehensive derivatization protocol among 17 tested reactions covering trifluoroacetylation, pentafluorobenzylation, methylations, and trimethylsilylations. MTBSTFA allowed easy and robust tert.-butyldimethylsilyl derivatization of 1-aminocyclopropane-1 carboxylic acid, indole-3-acetic acid, (+/-)-jasmonic acid, salicylic acid, (+/-) abscisic acid, meta-topolin, and trans-zeatin. Detection limits as analysed by selected ion monitoring quadrupole GC-MS were 0.2, 0.01, 1.0, 0.02, 0.3, 0.3, and 0.9 pmol of injected substance, respectively. Analysis of gibberellic acid A3, trans-zeatin riboside and (+/-)-abscisic acid-beta-D-glucopyranosyl ester was best when coupled by splitting extracts and trimethysilylation. The MTBSTFA derivatization protocol was optimised, and validated. The preparation was insensitive to 2% residual water and to < or = 1 day storage at room temperature. The final scheme was highly reproducible and successfully applied to extracts from approximately 300 mg (fresh mass) of tobacco (Nicotiana tabacum) root and Arabidopsis thaliana seedling. PMID- 12735442 TI - Determination of organophosphate esters in air samples by dynamic sonication assisted solvent extraction coupled on-line with large-volume injection gas chromatography utilizing a programmed-temperature vaporizer. AB - An on-line method for the determination of airborne organophosphate esters based on dynamic sonication-assisted solvent extraction and large-volume injection (LVI) gas chromatography with nitrogen-phosphorous detection is introduced. The LVI is performed with a programmed-temperature vaporizer. The entire extracted fraction of 800 microl (hexane-methyl-tert.-butyl ether, 7:3, v/v) is introduced directly into the GC system without any clean-up step following extraction. The extraction and analysis step were completed in less than 15 min. The limit of detection of the investigated organophosphate esters was established to be in the range of 5-32 pg/filter. The correlation coefficients (r2) were investigated in the linear range study of the entire system and established to be approximately 0.9900 for all the investigated organophosphates esters. Applications of the method was demonstrated with the extraction of air samples collected onto glass fiber filters from different indoor environments. Six organophosphate esters were found at the levels 0.4-138 ng/m3. PMID- 12735443 TI - Intercalibration of chromatographic methods for auxino phytodrugs in Solanaceae. AB - Three chromatographic methods are considered for the determination in Solanaceae of auxino-similar phytodrugs, so called because their structure resembles an auxine plant hormone. The phytodrugs studied were: 2,4-dichlorophenoxyacetic acid, 2,4-dichlorophenoxypropionic acid, 2,4-dichlorophenoxybutyric acid, 2 methyl-4-chlorophenoxyacetic acid, 2,4,5-trichlorophenoxyacetic acid, naphthylacetic acid and 2-naphthyloxyacetic acid. Three chromatographic methods, respectively based on ion-interaction HPLC, GC-MS with intra-injector derivatisation and CC-MS with pre-injection derivatisation, were developed, optimised and validated. A comparative discussion of the advantages/disadvantages of the methods suggests a strategy for their preferential use, that is essentially a function of the matrix complexity. PMID- 12735444 TI - Rapid analytical method for the determination of pesticide residues in sunflower seeds based on focused microwave-assisted Soxhlet extraction prior to gas chromatography-tandem mass spectrometry. AB - A rapid analytical method for determination of organochlorine pesticide residues in sunflower seeds based on focused microwave-assisted Soxhlet extraction has been developed. The main factors affecting the extraction efficiency--namely microwave power, irradiation time, volume of extractant and number of cycles- were optimized by a two-level factorial fractional design. After extraction, a liquid-liquid extraction and a clean-up step including the use of Florisil macrocolumns were required prior to injection of the extracts into the chromatograph in order to isolate the pesticide residues from the lipid fraction of the original extract. The MS-MS ion preparation mode was selected due to the high sensitivity and selectivity it provides. Seed samples were used collected near a crop subjected to aerial pesticide application. Residues of hexachlorocyclohexane isomers and endosulfan were found in the seeds although they were not subjected to pesticide application, thus showing the spray-drift contamination. The validation of the proposed approach was carried out by comparison with the ISO 659-1988 reference extraction method obtaining similar, or even better efficiencies by the proposed approach. PMID- 12735445 TI - Pyrolysis-gas chromatography-mass spectrometry of cured phenolic resins. AB - A previously described pyrolysis device with a movable reaction zone was used in the present work for thermal degradation of cured novolac and resol resins. After the pyrolysis the usual major components were identified, as well as relatively low-volatile aromatic compounds which were revealed after their separation in a capillary chromatographic column. Among the latter components, for both types of cured resins, xanthene and its methyl derivatives were found. The isomers of bis(hydroxyphenyl)methane were found in considerable quantities only in the case of partially cured novolac resins which contained any linear sequences in their structure. PMID- 12735446 TI - Highly sensitive determination of a polymeric hindered amine light stabilizer in polypropylene by reactive thermal desorption-gas chromatography using nitrogen specific detection. AB - Highly sensitive and specific determination of trace amounts of a polymeric hindered amine light stabilizer (HALS) in polypropylene (PP) materials could be established by improving reactive thermal desorption-gas chromatography (RTD-GC) in the presence of an organic alkali, tetramethylammonium hydroxide. By using nitrogen-phosphorus detection, highly selective detection of the HALS-related components was attained. In addition, the use of a polar poly(ethylene glycol) separation column alleviated the adsorption of minor specific pyrolysis products. This modified RTD-GC method allowed the determination of the polymeric HALS (Mr 1900) in PP even for trace concentrations between 100 and 500 ppm, through observing selectively the characteristic products containing a tetramethylpiperidine moiety, which had been impossible to detect under the previous RTD-GC conditions using a non-polar separation column and conventional flame ionization detection. PMID- 12735447 TI - Separation of haloacetic acids in water by capillary zone electrophoresis with direct UV detection and contactless conductivity detection. AB - The separation of haloacetic acids (HAAs) in water by capillary zone electrophoresis with direct UV and contactless conductivity detection was investigated using phosphate, citrate, and borate buffers, and the experimental data were compared to simulation data predicted by a computational program known as PeakMaster. Good agreement between the experimental data and simulation data predicted by PeakMaster was found. Using the phosphate buffer or the citrate buffer and electrokinetic injection it was possible to quantitate HAAs at 0.1 ppm levels in water. PMID- 12735448 TI - Comparing micellar electrokinetic chromatography and microemulsion electrokinetic chromatography for the analysis of preservatives in pharmaceutical and cosmetic products. AB - In this study, separation and determination of nine preservatives ranging from hydrophilic to hydrophobic properties, which are commonly used as additives in various pharmaceutical and cosmetic products, by micellar electrokinetic chromatograpy (MEKC) and microemulsion electrokinetic chromatography (MEEKC) were compared. The effect of temperature, buffer pH, and concentration of surfactant on separation were examined. In MEKC, the separation resolution of preservatives improved markedly by changing the sodium dodecyl sulfate concentration. Temperature and pH of running buffers were used mainly to shorten the magnitude of separation time. However, in order to detect all preservatives in a single run in a MEEKC system, a microemulsion of higher pH was needed. The separation resolution was improved dramatically by changing temperature, and a higher concentration of SDS was necessary for maintaining a stable microemulsion solution, therefore the separation of the nine preservatives in MEEKC took longer than in MEKC. An optimum MEKC method for separation of the nine preservatives was obtained within 9.0 min with a running buffer of pH 9.0 containing 20 mM SDS at 25 degrees C. A separation with baseline resolution was also obtained within 16 min using a microemulsion of pH 9.5 which composed of SDS, 1-butanol, and octane, and a shorter capillary column at 34 degrees C. Finally, the developed MEKC and MEEKC methods determined successfully preservatives in various cosmetic and pharmaceutical products. PMID- 12735449 TI - Development of a capillary electrophoretic method for the analysis of amino acids containing tablets. AB - Ketosteril is an enteral medicinal product indicated for prevention and therapy in chronic renal insufficiency in connection with a low protein diet. Tablets of Ketosteril contain five essential amino acids like: Lys, His, Thr, Trp, Tyr and another five amino acids in the form of their hydroxy and keto analogues as calcium salts, that are: alpha-ketoleucine, alpha-ketoisoleucine, alpha ketovaline, alpha-ketophenylalanine and alpha-hydroxymethionine. The composition of Ketosteril tablets is routinely tested with three LC methods. Capillary electrophoretic method seems to be a good alternative for amino acids and their analogues determination in multicomponent pharmaceuticals because of short analysis time and the possibility to assay all components during a single run without any pretreatment. Electrophoresis was performed in 50 microm I.D. fused silica capillaries with 65 cm distance to the detector. Capillaries were installed in Waters Quanta 4000 electrophoretic equipment with a positive power supply and on-line UV detection at 214 nm. Separations were done in a buffer containing 40 mM Tris and 160 mM boric acid titrated with NaOH to pH 10. The method developed allows the separation of all investigated analytes with an efficiency of n = 230,000 and 20 min analysis time. The method was applied for determination of all components of Ketosteril in commercial tablets. PMID- 12735450 TI - Reversed-phase liquid chromatographic method for the determination of selected room-temperature ionic liquid cations. AB - The separation of selected 1-alkyl- and 1-aryl-3-methylimidazolium-based room temperature ionic liquid cations has been performed using reversed-phase high performance liquid chromatography with electrospray ionization mass detection. The RP-HPLC method development started with the selection of a column taking into account especially the resolution of low molecular congeners of the selected group. Mobile phase composition was optimized for peak resolution, sensitivity and high reproducibility of retention values. The results of the method development were applied to the determination of exemplary ionic liquid species present in the medium used in cytotoxicity studies. PMID- 12735451 TI - Determination of aluminum in environmental and biological samples by reversed phase high-performance liquid chromatography via pre-column complexation with morin. AB - Morin was used as a pre-column reagent for the determination of aluminum by RP HPLC with fluorescence detection. This method has been successfully applied to direct determination of trace Al in environmental and biological samples. The response was linear from 6 x 10(-9) to 6 x 10(-5) M with a detection limit of 2 x 10(-9) M. In addition, the different Al complexes with morin were separated by the proposed HPLC procedure and their coordination ratios were depicted by molar ratio method. The results showed that 1:1 and 2:1 Al-morin complexes formed. PMID- 12735452 TI - Retention index calculation without n-alkanes--the virtual carbon number. AB - For the fast gas chromatographic identification of separated components the retention index is still one of the most often used tools, although mass spectrometry is available in almost all analytical laboratories. For the calculation of the retention indices it is not necessary to use n-alkanes or any other homologous series. If the analyte contains some compounds, not necessarily belonging to a homologous series, with well-known retention indices those compounds can be used as index references and the index of the other compounds can be calculated as is done using n-alkanes. The only difference is that instead of the carbon number of the n-alkanes, virtual carbon numbers of the reference compounds should be used. The method of calculation, and the effect of this method of calculation on the reproducibility are discussed in this paper, and analyses of some halogenated compounds and marjoram oil are used as experimental examples. PMID- 12735453 TI - Headspace solid-phase microextraction in combination with gas chromatography-mass spectrometry for the rapid screening of organophosphorus insecticide residues in strawberries and cherries. AB - A headspace solid-phase microextraction (HS-SPME) method in combination with GC MS was used for the extraction and quantification of diazinon, fenitrothion, fenthion, parathion ethyl, bromophos methyl, bromophos ethyl and ethion. The method was developed using a 100-microm poly(dimethylsiloxane) fiber. The obtained results showed higher responses of the insecticides after addition of aliquots of water and solvent to the fruit samples. Calibration curves that were constructed for the analytes spiked into strawberry and cherry samples followed linear relationships with good correlation coefficients (R2 > 0.986). Linearity range was between 50 and 500 microg/kg and the precision was found to be lower than 15% when applying the optimized HS-SPME procedure to fruit samples. Limits of detection in both strawberry and cherry samples using GC-MS (selected ion monitoring mode) were below 13 microg/kg. Moreover, the HS-SPME method was applied to the analysis of fruit samples and compared with liquid-liquid extraction. Results obtained in this study were in good agreement with those obtained using liquid-liquid extraction demonstrating that the recommended procedure was a fast, accurate and stable sample pretreatment method obtaining good efficiency for the extraction of organophosporus insecticides from strawberries and cherries. PMID- 12735454 TI - Specific analyte-electrolyte additive interaction in transient isotachophoresis capillary electrophoresis. AB - While cationic surfactants are usually included in the separation electrolyte to reverse the electroosmotic flow, the presence of the surfactant may also offer a means of capillary electrophoresis (CE) separation selectivity control over the anionic analytes, especially those that are prone to ion-pairing interaction. For one such analyte anion, iodide, the formation of several ion association/partition products with cetyltrimethylammonium chloride (CTAC) was first discovered when optimizing (decelerating) iodide mobility (in order to achieve effective transient isotachophoretic stacking). At comparatively high concentrations of iodide (> or = 0.01 mM) and the cationic surfactant well above the critical micelle concentration (25 mM), an additional peak due to interactions with the CTAC micelle was recorded, with a UV absorption spectrum fairly different from those of both interacting partners and also the iodide monomeric surfactant ion pair. Never observed before in normal CE mode, this phenomenon is believed to have occurred due to the enrichment effect of the initial isotachophoresis state. PMID- 12735455 TI - Capillary electrophoretic determination of Ga(III) based on the formation of a heteropolyoxomolybdate complex. AB - A novel capillary electrophoretic (CE) method was developed for the determination of Ga(III). The so-called Anderson-type [GaMo6O24H6]3- complex was readily formed by the reaction of Mo(VI) with Ga(III) in 0.050 M monochloroacetate buffer (pH 2.0) and the precolumn complex-formation reaction was applied to the CE determination of Ga(III) with direct UV detection at 240 nm. The peak area was linearly dependent on the concentration of Ga(III) in the range of 5.0 x 10(-7) 5.0 x 10(-5) M. Owing to the high molar absorptivity of the Anderson anion, a detection limit of 2.0 x 10(-7) M (signal-to-noise ratio=3) was achieved. The advantage of the present method is that the presence of large excesses of Al(III) and In(III) does not cause interference. PMID- 12735456 TI - Comments on the early history of gas chromatographic methods for oil analysis. PMID- 12735457 TI - Detection of VX contamination in soil through solid-phase microextraction sampling and gas chromatography/mass spectrometry of the VX degradation product bis(diisopropylaminoethyl)disulfide. AB - A solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC MS) sampling and analysis method was developed for bis(diisopropylaminoethyl)disulfide (a degradation product of the nerve agent VX) in soil. A 30-min sampling time with a polydimethylsiloxane-coated fiber and high temperature alkaline hydrolysis allowed detection with 1.0 microg of VX spiked per g of agricultural soil. The method was successfully used in the field with portable GC-MS instrumentation. This method is relatively rapid (less than 1 h), avoids the use of complex preparation steps, and enhances analyst safety through limited use of solvents and decontamination of the soil before sampling. PMID- 12735459 TI - Modeling of the salt effects on hydrophobic adsorption equilibrium of protein. AB - A two-state protein model is proposed to describe the salt effects on protein adsorption equilibrium on hydrophobic media. This model assumes that protein molecules exist in two equilibrium states in a salt solution, that is, hydrated and dehydrated states, and only the dehydrated-state protein can bind to hydrophobic ligands. In terms of the two-state protein hypothesis and the steric mass-action theory, protein adsorption equilibrium on hydrophobic media is formulated by a five-parameter equation. The model is demonstrated with the adsorption of bovine serum albumin to Phenyl Sepharose gels as a model system. The effects of salt type (sodium chloride, sodium sulfate and ammonium sulfate) on the model parameters are discussed. Then, the model formulism is simplified in terms of the small magnitude of the protein dehydration equilibrium constant in the model. This simplification has returned the model derived on the basis of the two-state protein hypothesis to its original mechanism of salt effects on the hydrophobic adsorption of protein. This simplified model also creates satisfactory prediction of protein adsorption isotherms. PMID- 12735458 TI - Advances in methodology for the validation of methods according to the International Organization for Standardization. Application to the determination of benzoic and sorbic acids in soft drinks by high-performance liquid chromatography. AB - Robust chemometric techniques such as least median of squares regression, H15 Huber estimator and Lenth's method are fundamental tools in the validation of analytical methods since they contribute the strategies needed to estimate efficiently parameters such as robustness, linear range, selectivity, accuracy (trueness and precision) and the capability of detection. In addition, the capability of discrimination defined as a generalisation of the capability of detection for any nominal concentration is evaluated. The new strategy proposed is applied to the validation of a chromatographic method for use in systematic analysis. PMID- 12735460 TI - Reproducible preparative liquid chromatography columns. AB - The level of reproducibility for sets of preparative liquid chromatography (prep LC) columns was studied using a self-packing axial compression system. Standard deviations of less than 5% of the mean of the column efficiency and less than 2% of the mean of the retention time are reported for three sets of packed, prep-LC columns. PMID- 12735461 TI - New ligand exchange chiral stationary phase for the liquid chromatographic resolution of alpha- and beta-amino acids. AB - A new ligand exchange chiral stationary phase (CSP) has been developed by covalently bonding (R)-N,N-carboxymethyl undecyl phenylglycinol mono-sodium salt onto silica gel and applied in the resolution of alpha- and beta-amino acids. In the resolution of alpha-amino acids, the new CSP was better insome cases than the old one, which was previously developed by covalently bonding (S)-N,N carboxymethyl undecyl leucinol mono-sodium salt onto silica gel, but worse in some other cases than the old one in terms of the separation factors (alpha). However, the new CSP wasalways much better than the old one in terms of the resolution factors (Rs). In the resolution of beta-amino acids, the new CSP was always much better than the old one in terms of both the separation and resolution factors. In an effort to characterize the new CSP, the chromatographic behaviors for the resolution of selected alpha- and beta-amino acids were investigated with the variation of the content of organic modifier and Cu(II) concentration in aqueous mobile phase and the column temperature. PMID- 12735462 TI - Synthesis and evaluation of a C8 stationary phase on a silica hydride surface by hydrosilation of 1-octyne. AB - The silanization/hydrosilation method is used to bond an alkyne (1-octyne) to a silica hydride surface. The new bonded material is characterized by elemental analysis and diffuse reflectance infrared Fourier transform spectroscopy. The hydrophobic behavior of this material was determined by the retention characteristics of aromatic solutes and the shape selectivity as well as phase classification (monomeric or polymeric) was measured by the polycyclic aromatic hydrocarbon mixture standard reference material 869. The presence of residual silanols on the bonded phase was probed by several basic solutes at pH 7. Long term stability studies were conducted by measuring retention and peak symmetry of basic compounds over several thousand column volumes at pH 10. PMID- 12735463 TI - Characterisation of legumes by enzymatic hydrolysis, microdialysis sampling, and micro-high-performance anion-exchange chromatography with electrospray ionisation mass spectrometry. AB - An assay based on enzymatic hydrolysisand microdialysis sampling, micro-high performance anion-exchange chromatography (micro-HPAEC) with electrospray ionisation mass spectrometry (ESI-MS) for the characterisation of legumes is presented. Characterisation of two bean varieties; Phaseolus mungo and P. acutifilous was based upon enzymatic hydrolysis using an endo-beta-mannanase from Aspergillus niger with subsequent analysis of the hydrolysates with HPAEC-MS. The hydrolysates were detected in the positive ionisation mode after desalting the chromatographic effluent, employing a cation-exchange membrane desalting device with water as the regenerating liquid. Mass chromatograms, acquiredafter hydrolysis of both bean samples for 12 h, showed two different profiles of hydrolysates. The P. mungo bean hydrolysate showed the presence of saccharides with a degree of polymerisation (DP) in the range of 2-6, whereas that of P. acutifilous showed only DPs of 2-5. Both bean samples had one type of DP 2, but showed different types of DPs 3, 4 and 5. Only the P. mungo sample showed the presence of DP 6. The most abundant fraction for P. mungo was DP 4, whereas that for P. acutifilous was DP 5. Tandem MS of the hydrolysates showed that the DP 2 hydrolysates observed for the samples were of the same type, having a 1,6 linkage. Also tandem MS data for DPs 3, 4, and 5 showed that similar hydrolysates were present within the same sample as well as among the two samples. The data also showed the existence of 1,6 linkages for DP 3, 4, and 5 hydrolysates. The single enzymatic hydrolysis in combination with microdialysis and HPAEC with ESI MS proved to be sufficient and reproducible for profiling and showing the difference between the two bean samples. PMID- 12735464 TI - Fractionation of starch hydrolysates into dextrins with narrow molecular mass distribution and their detection by high-performance anion-exchange chromatography with pulsed amperometric detection. AB - Low levels of high-molecular-mass dextrins in starch hydrolysates can be detected by high-performance anion-exchange chromatography with pulsed amperometric detection in spite of their low responses by dialysis of the starch hydrolysate and fractionation of the resulting adialysate with ethanol (final concentration 30-80% at 6 degrees C). In doing so, dextrin fractions with a relatively narrow molecular mass distribution were obtained. PMID- 12735465 TI - Use of liquid chromatography-diode-array detection and mass spectrometry for rapid product identification in biotechnological synthesis of a hydroxyprogesterone. AB - In exploratory scale biotechnological process development, the product must be rapidly identified although a reference compound may not always be available. LC diode-array detection and MS were used for this purpose in a process producing 9alpha-hydroxyprogesterone from progesterone as substrate. The electrospray ionization mass spectrometer was combined with an ion trap mass spectrometer for the second generation MS. The preliminary identification, which could be carried out within the course of a day, confirmed that the product was a hydroxyprogesterone. The final identification step, which was much more material intensive and hence time consuming, involved a two-step preparative separation to yield quantities necessary for definitive product identification based on 1H- and 13C NMR. PMID- 12735467 TI - In vitro comparison of complementary interactions between synthetic linear/branched oligo/poly-L-lysines and tissue plasminogen activator by means of high-performance monolithic-disk affinity chromatography. AB - The recently discovered serine protease called tissue plasminogen activator (t PA) enables efficient dissolution of blood clots. t-PA works by converting plasminogen into its active form, plasmin, dissolving the major component of blood clots, fibrin. The activation of plasminogen by t-PA is enhanced by the presence of fibrin, and this is probably due to the fact that both plasminogen and t-PA possess high affinity binding sites for fibrin. Besides fibrin, fibrin monomers and some fibrin(ogen) degradation products, certain synthetic polymers (for instance, poly-L-lysines) can provide the same stimulation of plasminogen activation. The recently developed high-performance monolithic-disk chromatography, HPMDC, could become the most convenient way to study biological pairs of interest. The inherent speed of HPMDC isolation facilitates the recovery of a biologically active product, since the exposure to putative denaturing influences, such as solvents or temperature, is reduced. The better mass transfer mechanism (convection rather than diffusion) allows to consider only the biospecific reaction as time limiting. The step-by-step modeling of hypothetical affinity pairs between t-PA and different types of oligo/polymer forms of linear and branched lysine derivatives obtained both by initiated polycondensation and solid-phase peptide synthesis using HPMDC seemed to be possible and a quite useful tool. The results of quantitative evaluation of such affinity interactions were compared with those established for natural affinity counterparts to t-PA (monoclonal antibodies, plasminogen, fibrinogen). The role of steric structure of lysine ligands was observed and analyzed. The results allowing to make the practical choice of affinity systems will be used for development of fast and efficient analytical and preparative methods for the downstream processes of recombinant production of this valuable enzyme. PMID- 12735466 TI - Determination of 3-nitrobenzanthrone in surface soil by normal-phase high performance liquid chromatography with fluorescence detection. AB - A sensitive method for determining 3-nitrobenzanthrone in surface soil was developed. 3-Nitrobenzanthrone was reduced to 3-aminobenzanthrone by refluxing at 60 degrees C with hydrazine and Raney nickel for 20 min, and 3-aminobenzanthrone was determined by normal-phase high-performance liquid chromatography (HPLC) with fluorescence detection. We used a cyanopropyl stationary phase and an n-hexane ethyl acetate (3:1, v/v) mobile phase, since 3-aminobenzanthrone exhibits fluorescence in a low-polarity solvent such as n-hexane or ethyl acetate, but not in a polar solvent such as water or methanol. The calibration graph showed good linearity (r2>0.9999) in the range of 0.002-2 ng, and the detection limit was 0.002 ng (S/N=3). 3-Nitrobenzanthrone in extracts from surface soil collected in the Chubu area (central area) of Japan was determined after clean-up using silica gel chromatography and high-performance liquid chromatography on a pyrenylethyl stationary phase. The concentration of 3-nitrobenzanthrone in surface soil was determined in the range of 1.2-1020 pg/g soil. PMID- 12735468 TI - Multiresidue method for fourteen fungicides in white grapes by liquid-liquid and solid-phase extraction followed by liquid chromatography-diode array detection. AB - A quantitative, selective and sensitive HPLC method for the analysis of 14 fungicides in white grapes for vinification is described. The proposed method is based on liquid-liquid extraction (LLE) and solid-phase extraction (SPE) followed by liquid chromatography and diode array detection (HPLC-DAD). Dichloromethane acetone (75:25, v/v) was the most appropriate solvent mix for extracting fungicides in white grapes. Silica cartridges resulted the most appropriate for extract purification purposes. Quality parameters of the proposed multiresidue method presented good recovery (ca. 85% for almost all target compounds) and precision (between 1.5 and 16%), and detection limits lower than maxima residual limits set by the 76/895/ECC and 90/642/ECC Directive. Five different white grapes for vinification produced in Rias Baixas area in Galicia (NW Spain) were analyzed in order to assess the performance of the method with real samples and to determine whether the concentration of the pesticides used exceed their maxima residue levels (MRLs). Results showed that grape concentrations for those identified fungicides were lower than those established by European legislation. PMID- 12735469 TI - Determination of abamectin and azadirachtin residues in orange samples by liquid chromatography-electrospray tandem mass spectrometry. AB - A rapid and sensitive LC-ESI-MS-MS method has been developed for the determination of azadirachtin and abamectin residues in orange samples. Samples were extracted with acetonitrile, in a high-speed blender. After the addition of sodium acetate, an aliquot of extract was directly injected into the LC-ESI-MS-MS system. The highest sensitivity of the method was achieved under MS-MS conditions using [M+Na]+ adducts as precursor ions. Recoveries for both compoundsfrom spiked orange samples at 0.01 and 0.1 mg/kg were above 80%, with good repeatability (<10%). Method detection limits achieved (<0.007 mg/kg) were adequate for the determination of these pesticides in this kind of sample from the regulatory point of view. The importance of the solvent used for extraction, as well as the addition of sodium acetate to the extracts and the selection of adequate chromatographic conditions are discussed. PMID- 12735471 TI - Comparison of Chirasil-DEX CB as gas chromatographic and ULMO as liquid chromatographic chiral stationary phase for enantioseparation of aryl- and heteroarylcarbinols. AB - For a broad spectrum of simple chiral alcohols, incorporating a (substituted) (het)aryl building block, enantiomer separation characteristics are reported for both gas chromatography on a Chirasil-DEX phase, and liquid chromatography on an (S,S)-ULMO phase. On this chiral Pirkle-type phase, homochiral enantiomers (mostly R) are eluted first without exception. The elution order R before S appears conserved as a rule also for gas chromatographic separations on Chirasil DEX, though with some remarkable exceptions indicating a change in the dominant discriminative mechanism. This was shown in the homologous series 1-phenylethanol to 1-phenylhexanol having the point of reversal at C4, while the o-methoxy analogues elute from C1 to C4 already in the reversed order. PMID- 12735470 TI - Liquid chromatography-mass spectrometry of spiroketal stereoisomers of pectenotoxins and the analysis of novel pectenotoxin isomers in the toxic dinoflagellate Dinophysis acuta from New Zealand. AB - The acid-catalyzed inter-conversion of spiroketal isomers of pectenotoxins PTX1, PTX6 and PTX2 were studied by liquid chromatography-electrospray ionization tandem mass spectrometry (LC-MS-MS). Using a C8-silica reversed-phase column and a mobile phase of aqueous acetonitrile containing 2 mM ammonium formate and 50 mM formic acid, the known spiroketal stereoisomers of PTX1 eluted in order of PTX1, PTX4 and PTX8, while those of PTX6 eluted in the order PTX6, PTX7 and PTX9. Acid treatment of PTX2 yielded two novel spiroketal stereoisomers, which have been named PTX2b and PTX2c. LC-MS-MS spectra obtained for the [M+NH4]- ions of PTX2, PTX2b and PTX2c were essentially identical. As an application of the LC-MS-MS methodology, a sample of the toxic dinoflagellate Dinophysis acuta collected from the coast of New Zealand was analyzed for pectenotoxins. PTX2 and a new pectenotoxin, which has been named PTX11, were detected as the most predominant compounds. Novel PTX2 and PTX11 isomers were also found in the D. acuta although the levels of these compounds were low. PMID- 12735472 TI - Further improvements in the application of high-performance liquid chromatography, capillary electrophoresis and capillary electrochromatography to the analysis of algal toxins in the aquatic environment. AB - The presence of algal toxins in the aquatic environment represents an important socioeconomic concern in many places worldwide, due to the toxicity that these compounds can induce in seafood or freshwater organisms at very low levels. Several analytical alternatives have been proposed over the last years for the control of these contaminants, which acute or chronic toxicity requires low detection levels and demands for the search of sensitive methods for their detection and determination. HPLC has been widely used for this purpose, although several alternatives such as CE or capillary electrochromatography (CEC) are being lately developed with this aim. In this work we report on the application of improved HPLC, as well as CE and CEC, for the analysis of diarrhetic shellfish poisoning toxins, amnesic shellfish poisoning (ASP) toxins and microcystins (MCs) present in different matrices such as water, shellfish or algae. Improvements in sample preparation for increasing sensitivity and selectivity are also shown. While UV and fluorimetric detection are the detection methods generally used, mass spectrometric detection was also applied for ASP toxins and MCs, especially for confirmatory purposes. From the results obtained it can be concluded that both HPLC and CE offer a good potential for a sensitive and selective determination of these algal toxins in such complex matrices. The results obtained for CEC allow also to conclude that this technique can result in a promising technique for such application. PMID- 12735474 TI - Determination of carbohydrates as their 3-aminophthalhydrazide derivatives by capillary zone electrophoresis with on-line chemiluminescence detection. AB - A method based on pre-capillary derivatization with luminol (3 aminophthalhydrazide) for carbohydrate analysis using capillary electrophoresis with on-line chemiluminescence (CL) detection was developed. The derivatives of seven monosaccharides were separated and detected by using 200 mM borate buffer containing 100 mM hydrogen peroxide at pH 10.0 as separation electrolyte and 25 mM hexacyanoferrate in 3 M sodium hydroxide solution as post-capillary chemiluminescence reagent with separation efficiencies ranging from 160,000 to 231,000 plates per metre. The minimum amount of carbohydrate derivatized was 2 pmol (corresponding to the concentration of 2 microM). The method also provided a linear response for glucose in the concentration range of 0.1-250 microM with a mass detection limit of 420 amol or a concentration detection limit of 0.1 microM. Preliminary work using the CE-CL format to determine glucose in a rat brain microdialysis sample is presented as a typical case. PMID- 12735473 TI - Advantages and limitations of coupling isotachophoresis and comprehensive isotachophoresis-capillary electrophoresis to time-of-flight mass spectrometry. AB - Capillary isotachophoresis (ITP) and comprehensive isotachophoresis-capillary electrophoresis (ITP-CE) were successfully coupled to electrospray ionization (ESI) orthogonal acceleration time-of-flight mass spectrometry (TOF-MS) using angiotensin peptides as model analytes. The utility of ITP-TOF-MS and ITP-CE-TOF MS for the analysis of samples containing analyte amounts sufficient to form flat top ITP zones (30 microM) as well as for samples with trace analyte amounts (0.3 microM) was studied. Separations were performed in 150 microm internal diameter (I.D.) capillaries for the ITP experiments, and in 200 microm I.D. (ITP) and 50 microm I.D. (CE) capillaries for ITP-CE experiments. The fused-silica columns were coated with poly(vinyl alcohol) to suppress electroosmotic flow that can disrupt ITP zone profiles. The sample loading capacity in both ITP and comprehensive ITP-CE was greatly enhanced (up to 10 microl) compared with typical nanoliter-sized injection volumes in CE. It was concluded that ITP-TOF-MS alone was adequate for the separation and detection of high concentration samples. The outcome was different at lower analyte concentrations where mixed zones or very sharp peaks formed. With formation of mixed zones, ion suppression and discrimination could occur, complicating quantitative determination of the analytes. This problem was effectively overcome by inserting a CE capillary between the ITP and TOF-MS. In such an arrangement, samples were preconcentrated in the high load WTP capillary and then injected into a CE capillary where they were separated into non-overlapping peaks prior to their detection by TOF-MS. The advantage of this comprehensive arrangement, which we have described previously, is that there is no need to discard portions of the sample in order to avoid overloading of the CE capillary. The whole sample is analyzed by multiple injections from ITP to CE. Thus, this method can be used for the analysis of complex samples with wide ranges of component concentrations. PMID- 12735475 TI - Preparative isolation and purification of two isoflavones from Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao by high-speed counter-current chromatography. AB - Two isoflavones, calycosin-7-O-beta-D-glycoside and formononetin-7-O-beta-D glycoside, were separated from n-butanol extract of the root of Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao by high-speed counter-current chromatography in two steps using two different solvent systems composed of ethyl acetate-ethanol-n-butanol-water (30:10:6:50, v/v) and ethyl acetate-ethanol-water (5:1:5, v/v). From 200 mg of crude extract, calycosin-7-O-beta-D-glycoside (12 mg) and formononetin-7-O-beta-D-glycoside (10 mg) were isolated at over 95% purity by HPLC analyses, and their structures were identified by MS, 1H NMR and 13C NMR. PMID- 12735476 TI - Changes in the order of elution of some organic compounds in high-resolution gas chromatography on polar columns depending on the injection method and water content in the sample. AB - During the GC analysis of apple aroma essence using a capillary column with polar stationary phases, i.e., bonded/crosslinked polyethylene glycol (HP-Innowax) and acid-modified polyethylene glycol (FFAP) the phenomenon of changing the elution order of some compounds was observed ("cross-over phenomenon"). The elution order within two pairs of closely eluting compounds: 2-methylbutyl acetate/pentan-3-ol (internal standard) and trans-2-hexen-1-al/2-methylbutan-1-ol differed between samples introduced onto the column with direct injection and with static headspace method. Experiments carried out on standard solutions of investigated compounds in water-acetone mixtures of gradually reduced water ratio (99:1 to 0:100) proved that changes of elution times leading first to co-elution and subsequently to reversion of elution order of compounds within both pairs depended on the amount of water in the injected sample. The same effect was observed while the amount of water was diminished by reducing the sample volume introduced into the column. PMID- 12735477 TI - High fructose feeding of magnesium deficient rats is associated with increased plasma triglyceride concentration and increased oxidative stress. AB - The purpose of this study was to assess whether dietary carbohydrate could differentially influence the consequences of magnesium deficiency with particular emphasis on lipid metabolism and oxidative stress. Rats were fed a sucrose based or starch based diet either adequate or deficient in magnesium for two weeks. Magnesium deficient rats, as compared with rats fed magnesium adequate diets, displayed the usual decrease in plasma magnesium concentration. The classic symptoms of inflammation including hyperaemia, increased number of blood leukocytes and enlarged spleen weight were observed in these rats. Plasma TG and plasma apo B concentrations were also significantly increased. In addition, magnesium-deficient animals presented an increased susceptibility to lipid peroxidation of heart and liver tissues as shown by TBARS concentration. Regardless of magnesium status, sucrose feeding did not affect the magnesium plasma level and inflammatory parameters. Feeding rats the sucrose diets induced hypertriglyceridaemia and increased plasma apo B concentration. Heart and liver susceptibility to lipid peroxidation were significantly increased in rats fed the sucrose diets as compared with those fed the starch diets. Sucrose feeding in magnesium deficient rats was associated with higher plasma triglycerides concentration and higher tissue susceptibility to peroxidation as compared with magnesium deficient rats fed the starch diet. The results emphasised the potential detrimental and additional effect of sucrose feeding and magnesium deficiency on cardiovascular risk. Since the intake of magnesium has been reduced appreciably in industrialised countries while fructose consumption has been rapidly increased, the impact of this eating pattern should be clarified in humans. PMID- 12735478 TI - Effect of magnesium supplementation on oxidative stress in alloxanic diabetic rats. AB - Magnesium deficit and oxidative stress are common features of the diabetic state. This concept supported by another observation that magnesium deficiency is also a state of increased oxidative stress prompted us to study the effect of magnesium supplementation on magnesium status and oxidative stress in diabetic rats. For this purpose, male Wistar rats were made diabetic with a single intraperitoneal injection of Alloxan. Experimental diabetes caused a significant decrease in serum and red blood cell magnesium levels and increased urinary excretion of magnesium. Marked increase in plasma malondialdehyde and corresponding decrease in vitamins C & E, uric acid and total thiols was observed in the diabetic rats as compared to control group. In liver, MDA levels were increased significantly with concomitant decrease in vitamin C, non-protein thiols and antioxidant enzymes (SOD & GST). Magnesium supplementation for four weeks restored serum and RBC magnesium levels to near normal levels with marginal but significant decrease in blood glucose levels. Plasma and liver MDA levels were reduced significantly and vitamin C and total thiols were increased significantly with magnesium supplementation. Antioxidant enzyme activity was also increased significantly with magnesium supplementation in diabetic rats. Our data clearly demonstrates that alloxanic diabetes is associated with decreased magnesium status and increased oxidative stress and that magnesium supplementation can in part restore the antioxidant parameters and decrease the oxidative stress in experimental diabetic rats. PMID- 12735479 TI - Magnesium deficiency reveals the neurotoxicity of delta-9-tetrahydrocannabinol (THC) low doses in rats. AB - In the present study, muricide behaviour (MB) was studied in Long Evans rats in various situations. The MB pattern of each experimental group was compared, in 6 successive assays 1 hr-delayed to that of natural killer rats (NK). The percentage of NK rats was 11% in the strain used. In the 11 mg THC/kg b.w. treated naive rats, a significant additional percentage of rats (59%) became muricidal. The durations of the 3 MB phases were significantly increased as a result of an increased aggressiveness in the 1st assay but returned progressively to NK values on the 6th assay, in parallel with the physiological elimination of THC. This result indicates a true killing training in those non killer rats that became muricidal under THC. A severe magnesium deficiency induced by a 50 ppm magnesium-deficient diet induced 100% MB whereas a 150 ppm magnesium deficiency did not induce additional MB. In the severe deficiency, the MB pattern was rather similar to that of NK with the exception of the attack on the living mouse which was doubled probably because of magnesium-induced hyperexcitability responsible for a lower attack efficiency. In both 50 but also 150 ppm magnesium-deficient rats, a single injection of THC at low doses (2, 4 or 8 mg THC/kg b.w.) which is without aggressive effect in control rats, induced a 100% MB, the pattern of which was all the more severe as the magnesium deficiency was important or the THC dose higher. The pattern showed an important decrease in the two first phases and a dramatic increase in the attack on the dead mouse, suggesting that the combination of both treatments provoked severe central damage with a compulsive killing behavior. Consequently, it appears that a magnesium deficiency, even a moderate one, may aggravate the neurotoxicity of THC at low doses and, reciprocally, that low doses of THC may reveal the potential neurotoxicity of a moderate magnesium deficiency. PMID- 12735480 TI - Comparison of a short irradiation (50 sec) by different wavelengths on audiogenic seizures in magnesium-deficient mice: evidence for a preventive neuroprotective effect of yellow. AB - Audiogenic seizures triggered by an acoustic stimulus of determined frequency and amplitude have been described in many laboratory animals in many circumstances including magnesium deficiency. This model, recently validated, was used, in DBA/2 mice, to study the preventive neuroprotective effect of 6 wavelengths of the visible spectrum used in Chromatotherapia* (lambda(max) 440, 484, 528, 572, 616 and 660 nm) at low irradiance. Each short illumination lasted 50 seconds and was followed by 20 minutes of darkness. It appeared that yellow fully protected 16 out of 17 mice from seizure occurrence. Green allowed the survival of 69% of mice but did not protect them from seizure occurrence. On the contrary, the other four colors (orange, red, purple and blue) failed to protect the mice and showed a tendency to accelerate their death. White color was not protective but allowed the difficult survival of 30% of mice. Darkness had no protective effect. These results even though surprising open a great field of investigation. PMID- 12735481 TI - Physiological importance of the connective tissue in the human amnion. Role of magnesium. AB - The elemental ionic distribution in the epithelial layer (EL) and in connective tissue (CT = compact layer + fibroblast layer) of the human amniotic membrane has been studied in reference samples, after conservation in a physiological fluid (Hanks' solution) and after addition of 2 mM MgCl2 in Hanks' solution. Particle induced X-ray emission and Rutherford backscattering spectrometry techniques were used to provide quantitative measurements. In physiological fluid, with regard to reference samples, the monovalent ions (Na+, K +, Cl-) concentrations were identical on both layers. This data indicates that the connective tissue, in particular the compact layer, acts as a buffer which fix minerals. Mg2+ and Ca2+ levels were higher in EL than in CT. The addition of MgCl2 in Hanks' solution induced a decrease of the monovalent ion concentrations in both layers except Na+ level in EL which remained constant, an increase of the Mg2+ level in both layers, while the Ca2+ remained constant. These data indicate the possible role of connective tissue in pregnancies complicated by poly or oligohydramnios. PMID- 12735482 TI - Structural alterations of the vascular wall in magnesium-deficient mice. A possible role of gelatinases A (MMP-2) and B (MMP-9). AB - Vascular alterations during magnesium deficiency have long been known but the implicated mechanisms have so far been poorly documented. In this preliminary assay, we compared the thoracic aortic histology in Swiss OF1 mice fed a severe magnesium-deficient diet (50 +/- 5 ppm) for 42 days to that of controls fed a standard diet (1700 +/- 100 ppm magnesium). It appeared (eosin-haematoxylin coloration) that, in magnesium-deficient mice, the aortic wall was thinner than in controls. Specific colorations of the two of main fibers vascular tissue (collagens and elastin) showed severe structural alterations of both components. These changes were consecutive to the expression of matrix metalloproteinases (MMP) -2 and -9 which were present as zymogens (inactive forms) in controls and supposed to be present in their active and inactive forms in magnesium-deficient mice (zymography). These changes which have not been reported so far would explain, at least in part, the sensitivity of magnesium-deficient mice to various stress or xenobiotics. PMID- 12735483 TI - Effect of magnesium on granulocyte function and on the exercise induced inflammatory response. AB - Magnesium status is a well-known modulator of the immune system. In the present study we investigated the effect of magnesium on granulocyte signalling and function. Furthermore, we performed a double-blinded randomised study investigating the effect of a two-month magnesium supplementation period on the exercise-associated alterations in immune function. In vitro incubation of granulocytes in media of different magnesium composition resulted in significant changes in chemotactic peptide-induced calcium transients while basal calcium levels were not affected. Likewise, the stimulus-induced formation of free radicals was affected by extracellular magnesium while phagocytosis of granulocytes was not affected. In the second part of the study we investigated whether a two-month period of magnesium supplementation was able to diminish alterations in immune cell counts and functions after an exercise test until exhaustion. The magnesium status was similar in both human and placebo groups and did not change significantly after the supplementation period. Exhaustive exercise induced an activation of the immune system as indicated by an increase in granulocyte count and a post-exercise lymphopenia. In addition, chemotactic peptide-induced cellular calcium transients were enhanced post-exercise while oxidative burst and phagocytosis were decreased. These results suggest that magnesium is an important modulator of immune cell function under in vitro conditions. However, a magnesium supplementation seems to be unable to prevent any exercise-associated alterations in immune cell function in athletes with balanced magnesium status. PMID- 12735484 TI - Marfan syndrome, magnesium status and medical prevention of cardiovascular complications by hemodynamic treatments and antisense gene therapy. AB - The medical management of Marfan Syndrome (MFS) mainly relies on early prevention of the aortic complications. Hemodynamic treatments try to diminish the forcefulness of cardiac contractions and to reduce blood pressure: for example long term administration of propranolol may significantly reduce the rate of increase in aortic ratio (aortic diameter/expected aortic diameter). Retardation of aortic dilatation may be most often observed by early treatment started when the baseline end-diastolic aortic root diameter is < 40 mm. It seems better to use beta-blockers without intrinsic sympathomimetic activity. Successful acceptance of beta-blockers may be limited by side-effects, but the efficiency of alternative hypotensive agents (calcium channel inhibitors, ACE inhibitors) is not yet validated. Gene therapy might constitute an etiologic specific treatment of MFS. FBN1-RZ1 hammerhead antisense ribozyme is able to suppress expression of the mutant FBN1 allele. The use of ribozymes as systemic therapeutic agents will depend on efficient delivery to its target, but the various proposed vectors raise yet unsolved problems. A hydrogel angioplasty balloon might be a possible vector for delivering an antisense ribozyme in the aortic wall specifically. Ribozymes--as deoxyribonucleotides--may be taken up by tissue upon local application. Further research should study ex vivo local application of antisense ribozyme on human aortic wall, before assessing in vivo efficiency and tolerance of this aortic local vectorisation. It is always necessary to maintain a balanced magnesium intake in patients with MFS. Firstly to prevent the multiple noxious effects of magnesium deficiency on cardiovascular targets. Secondly to ensure the best efficiency and the least toxicity of the hemodynamic drugs used as long term prophylactic treatment for cardiovascular complications and of the etiologic antisense magnesium-dependent gene therapy, in the future. PMID- 12735485 TI - Intravenous magnesium sulphate in acute myocardial infarction--is the answer "MAGIC"? AB - The role of magnesium in coronary artery disease has been evaluated extensively during the last three decades. The intravenous application of magnesium in acute myocardial infarction is of major importance, the beneficial effects have been underlined in several studies. Magnesium is of significance in the pathomechanisms of reperfusion injury and reduction of malign arrhythmias in the critical acute phase of myocardial infarction, if applied intravenously. However, the promising results of LIMIT-2 could not be confirmed by the data of ISIS-4. The timing of magnesium therapy is probably the most important key factor. Similar to the guidelines of thrombolytic intervention, magnesium has to be administered as early as possible, at the latest before myocardial reperfusion has started. Nevertheless, because of conflicting results of prior trials doubts on the efficacy of intravenous magnesium in myocardial infarction still remain. The multinational, multicenter trial MAGIC has been set up to evaluate the optimal patient cohort as well as the ideal dose regimen for the application of intravenous magnesium sulphate in patients with acute myocardial infarction. The answer on the open questions on intravenous magnesium sulphate in myocardial infarction could be "MAGIC". PMID- 12735486 TI - Magnesium and connective tissue. AB - Magnesium (Mg2+) is the fourth most abundant cation and the second most abundant intracellular cation in vertebrates. Magnesium plays a critical role in cellular functions such as: cell adhesion via integrins on various macromolecular substrates, cell migration, DNA transcription and protein synthesis. PMID- 12735487 TI - Paradoxical drug response and the placebo effect: a discussion of Grunbaum's definitional scheme. AB - Grunbaum claims that the remedial failure of a treatment's characteristic factors is the generic, objective property of a placebo. He stipulates that a treatment is placebic if this remedial failure exacerbates the target disorder. This stipulation can subsume as placebic effects that might be solely pharmacological, e.g., paradoxical reactions to certain psychiatric drugs. If that exacerbation can be explained pharmacologically, then we might question whether Grunbaum's definitional scheme captures the core identity of what we usually intend by the placebo concept. I propose that this core identity is best captured by a symbolic meaning hypothesis in which psychological factors exert the determinative influence. PMID- 12735488 TI - Pragmatism as a research program--a reply to Arras. AB - This paper is a reaction to an article by John Arras published earlier in this journal. In this article Arras argues that "freestanding pragmatism" has little new to offer to bioethics. We respond to some of Arras' arguments and conclude that, although he overstates his case at certain points, his critique is, broadly speaking, correct. We then introduce and discuss an alternative approach to pragmatist ethics, one which puts to work the ideas and insights of pragmatism conceived as a broad philosophical movement, without lapsing into a canon dependent approach. The approach we propose exhibits a number of characteristics that differ from Arras's account of freestanding pragmatism and offers something new to bioethics. PMID- 12735490 TI - Metaphors in behavioral genetics. AB - Behavioral geneticists sometimes use metaphors to describe the role of genes in human behavior. In this paper, five sample texts are analyzed: a popular book, a textbook, a scientific review article, and two original scientific articles representing different approaches in behavioral genetics. Metaphors are found in all the different kinds of sample texts, not only in the popular book and the textbook. This suggests that metaphors are used not only for rhetorical or pedagogical purposes but play a more fundamental role in scientific understanding. In the sample texts, the metaphors tend to be antideterministic, i.e., they do not imply genetic determinism but stress the interaction of multiple genes and multiple environmental factors. No conclusion can be drawn, however, as to whether antideterminism is representative of present-day behavioral geneticists in general. Certain historically important metaphors that may imply genetic determinism are qualified, avoided, or explicitly rejected. There are tensions between some of the metaphors, making them difficult to combine. All the metaphors that are used appear empirically apt, however sometimes only with certain qualifications. PMID- 12735489 TI - Gene therapy: ethical issues. AB - To discern the ethical issues involved in current gene therapy research, to explore the problems inherent in possible future gene therapies, and to encourage debate within the scientific community about ethical questions relevant to both, we surveyed American Society of Human Genetics scientists who engage in human genetics research. This study of the opinions of U.S. scientific experts about the ethical issues discussed in the literature on gene therapy contributes systematic data on the attitudes of those working in the field as well as elaborative comments. Our survey finds that respondents are highly supportive of the potential use of somatic cell gene therapy to cure serious diseases in adults and children as well as prospective offspring. A clear majority, however, believe that using such genetic techniques for enhancement purposes is unacceptable. Delineating the line between disease/disorder and improvement/enhancement poses a problem not easily resolved and one conducive to the growth of slippery-slope apprehensions. The majority of respondents also advocate germ-line therapy, in theory at least, and under similar restrictions, but they recognize the roadblock that the existence of unanticipated negative consequences currently presents. Another complex matter involves trying to determine appropriate reasons for choosing target diseases for research, for which the dichotomy between rare single-gene and common multifactorial diseases reveals an ongoing dilemma. PMID- 12735492 TI - Weighted estimating equations for linear regression analysis of clustered failure time data. AB - Estimation of regression parameters in linear survival models is considered in the clustered data setting. One step updates from an initial consistent estimator are proposed. The updates are based on scores that are functions of ranks of the residuals, and that incorporate weight matrices to improve efficiency. Optimal weights are approximated as the solution to a quadratic programming problem, and asymptotic relative efficiencies to various other weights computed. Except under strong dependence, simpler methods are found to be nearly as efficient as the optimal weights. The performance of several practical estimators based on exchangeable and independence working models is explored in simulations. PMID- 12735491 TI - On medicine as a human science. AB - All the powerful influences exerted by the subjective-interpersonal dimension on the organic or technical-functional dimension of sickness and health do not make an intersubjective test concerning medical therapeutic results impossible. These influences are not arbitrary; on the contrary, they obey "laws" that are de facto sufficiently stable to allow predictions and explanations similar to those of experimental sciences. While, in this respect, the rules concerning human action are analogous to the scientific laws of nature, they can at any time be revoked by becoming aware of them. Law-like and reproducible regularities in the sciences of man are by no means separated from a patient's personal-hermeneutic mediation. This makes it possible for human beings to modify, improve or sometimes even entirely (or better almost entirely) suspend these psychological, sociological, ethnological, medical, regularities. For this reason the sciences of man including medicine are under the obligation of constantly inspecting the continuing validity of the rules on which their predictions and explanations are based, namely by indirect, statistical methods. This requires a synergistic collaboration of extra-clinical and clinical tests through which medicine can obtain a good level of intersubjective testability. PMID- 12735493 TI - Maximum penalized likelihood estimation in a gamma-frailty model. AB - The shared frailty models allow for unobserved heterogeneity or for statistical dependence between observed survival data. The most commonly used estimation procedure in frailty models is the EM algorithm, but this approach yields a discrete estimator of the distribution and consequently does not allow direct estimation of the hazard function. We show how maximum penalized likelihood estimation can be applied to nonparametric estimation of a continuous hazard function in a shared gamma-frailty model withright-censored and left-truncated data. We examine the problem of obtaining variance estimators for regression coefficients, the frailty parameter and baseline hazard functions. Some simulations for the proposed estimation procedure are presented. A prospective cohort (Paquid) with grouped survival data serves to illustrate the method which was used to analyze the relationship between environmental factors and the risk of dementia. PMID- 12735494 TI - A discrete-time multilevel mixture model for event history data with long-term survivors, with an application to an analysis of contraceptive sterilization in Bangladesh. AB - Event history models typically assume that the entire population is at risk of experiencing the event of interest throughout the observation period. However, there will often be individuals, referred to as long-term survivors, who may be considered a priori to have a zero hazard throughout the study period. In this paper, a discrete-time mixture model is proposed in which the probability of long term survivorship and the timing of event occurrence are modelled jointly. Another feature of event history data that often needs to be considered is that they may come from a population with a hierarchical structure. For example, individuals may be nested within geographical regions and individuals in the same region may have similar risks of experiencing the event of interest due to unobserved regional characteristics. Thus, the discrete-time mixture model is extended to allow for clustering in the likelihood and timing of an event within regions. The model is further extended to allow for unobserved individual heterogeneity in the hazard of event occurrence. The proposed model is applied in an analysis of contraceptive sterilization in Bangladesh. The results show that a woman's religion and education level affect her probability of choosing sterilization, but not when she gets sterilized. There is also evidence of community-level variation in sterilization timing, but not in the probability of sterilization. PMID- 12735495 TI - Survivor function estimators under group sequential monitoring based on the logrank statistic. AB - In this paper we investigate a group sequential analysis of censored survival data with staggered entry, in which the trial is monitored using the logrank test while comparisons of treatment and control Kaplan-Meier curves at various time points are performed at the end of the trial. We concentrate on two-sample tests under local alternatives. We describe the relationship of the asymptotic bias of Kaplan-Meier curves between the two groups. We show that even if the asymptotic bias of the Kaplan-Meier curve is negligible relative to the true survival, this is not the case for the difference between the curves of the two arms of the trial. A corrected estimator for the difference between the survival curves is presented and by simulations we show that the corrected estimator reduced the bias dramatically and has a smaller variance. The methods of estimation are applied to the Beta-Blocker Heart Attack Trial (1982), a well-known group sequential trial. PMID- 12735496 TI - Mixed discrete and continuous Cox regression model. AB - The Cox (1972) regression model is extended to include discrete and mixed continuous/discrete failure time data by retaining the multiplicative hazard rate form of the absolutely continuous model. Application of martingale arguments to the regression parameter estimating function show the Breslow(1974) estimator to be consistent and asymptotically Gaussian under this model. A computationally convenient estimator of the variance of the score function can be developed, again using martingale arguments. This estimator reduces to the usual hypergeometric form in the special case of testing equality of several survival curves, and it leads more generally to a convenient consistent variance estimator for the regression parameter. A small simulation study is carried out to study the regression parameter estimator and its variance estimator under the discrete Cox model special case and an application to a bladder cancer recurrence dataset is provided. PMID- 12735497 TI - Contemporary pathology of prostate cancer. AB - In less than 20 years since the introduction of serum PSA and the spring-loaded 18-gauge prostatic biopsy needle, pathologists have adjusted to the limited tissue requirements of narrow needle specimens to apply criteria for diagnosis and grading of prostate cancer, borrowing from lessons learned from radical prostatectomies. Substantial gains have been made during this period in the understanding of precancerous lesions, mimics of malignancy, the criteria for minimal cancer, variants of cancer, and treatment-induced changes. The light microscopic findings remain the criterion standard for diagnosis against which all new techniques should be measured. Numerous findings have proven to be of value, including simple quantitation of histopathologic features, cancer volume, perineural invasion, and others. PMID- 12735498 TI - The epidemiology of prostate cancer. AB - The etiology of prostate cancer remains virtually unknown. Although there are a number of new leads with regard to risk factors for prostate cancer, more research is required to confirm them. There is little purpose in conducting further case-control studies of prostate cancer-particularly since the use of PSA testing has become wide-spread. Instead, future epidemiologic studies should focus on prostate tumor subclassification, in terms of method of detection, markers of biological "aggressiveness," and genetic changes. Many of these new leads involve the possible influence of polymorphisms in key genes involved in important physiologic processes in the prostate. To fully explore the complexity of interrelationships between the several elements in these pathways will require large cohort studies in which blood is sampled prior to diagnosis. Such studies will be important for identifying which modifiable aspects of lifestyle (such as diet, alcohol, tobacco, physical activity) might be targeted for intervention, to reduce risk. The detection of early prostate cancers by PSA testing relatives of men with prostate cancer has affected the prevalence of phenocopies and, hence, the meaningfulness of risk estimation in prostate cancer families. Because multiple-case families form the substrate for gene hunting via linkage analysis, this phenocopy phenomenon is going to cause considerable confusion and wasted effort. Presently, men with a family history of prostate cancer can be provided with little advice in terms of preventive action. It is likely that one or more genetic mutations associated with a high risk for prostate cancer will be identified in the near future. Even so, the risks probably will be similar to those for mutations in the first two breast cancer genes--informative for very few families. It is difficult to foresee, as and when high-risk mutation carriers are identified, what advice should be offered to them: prophylactic prostatectomies seem to have less attraction than do prophylactic mastectomies for women at high risk of breast cancer. This issue becomes more complex when considering counseling on the basis of a genetic profile involving many low-risk polymorphisms. Hopefully, such genetic screening should occur only after its efficacy has been established; when there is a better understanding of prostate biology, tumor heterogeneity, and prognosis; and when there are proven treatment or prevention options available. Prevention is held to be better than cure, and there are some potentially interesting chemopreventive agents that require careful trial before public health initiatives can be promoted. Potential candidates include vitamin E, selenium, zinc, and lycopene as dietary supplements. There are other agents that may be appropriate for pharmaceutical development, including inhibitors of COX-2 and IGF-1 activity. It is important that chemoprevention trials are followed-up for a sufficient period of time and that other endpoints also are captured, because the supplementation of diets with superphysiologic doses of individual micronutrients sometimes has caused unexpected and unwanted results--for example, an 18% increase in lung cancers observed in the beta-carotene arm of the ATBC trial. PMID- 12735499 TI - The natural history of prostate cancer. AB - Predicting the long-term outcome of patients who choose watchful waiting as initial therapy for prostate cancer is difficult. The wide variation in disease progression, the impact of competing medical hazards, and the potential impact of early hormonal therapy that is characteristic of contemporary patients all conspire to compromise survival estimates dating from the pre-PSA era. The survival analysis figure developed by Albertsen et al (Fig. 1) estimates a 15 year survival rate based on patient age and Gleason score at diagnosis from patients diagnosed in the pre-PSA era. Although no effort was made to adjustfor competing medical hazards, patients and clinicians can adjust a patient's chronological age to match his "physiological" age. The advent of widespread PSA testing appears to have advanced the date of diagnosis by approximately 5 years and the onset of secondary treatment by at least as many years. Therefore, the figure describing the natural history of prostate cancer most likely underestimates rather than overestimates survival among men with newly diagnosed, localized prostate cancer who select watchful waiting as their treatment choice. As contemporary databases of men with localized prostate cancer mature, more data on the natural history of this disease will become available. Only time will tell how the use of PSA has altered the precision of historic case-series data. PMID- 12735501 TI - Screening for prostate cancer. AB - This article provides a review of present evidence that suggests that screening achieves a reduction in prostate cancer mortality. Aspects of quality of life and cost effectiveness are reviewed, as well as present test performance. The questions addressed in this article include the following: Is screening in its present form acceptable as a public health instrument? What changes are needed to improve the screening procedure? What are the research priorities in this field, assuming that the present evidence of prostate cancer mortality reduction is eventually confirmed by ongoing randomized controlled trials? PMID- 12735500 TI - Chemoprevention of prostate cancer. Focus on key opportunities and clinical trials. AB - By 2004-2005, the final results of the Prostate Cancer Prevention Trial should be available. Within several years thereafter, results of the SELECT should be available. The growing list of potential agents for prostate cancer prevention continues to grow and includes COX-2 inhibitors, vitamin D, dietary interventions (soy, isoflavenoids, low-fat diet), and many others. As prospective trials are completed and molecular genetic correlations are developed, it is almost certain that specific recommendations, tailored to the individual patient, will be developed. Ultimately, through these efforts, it can be anticipated that the primary focus for control of prostate cancer will shift from early detection and treatment to prevention. PMID- 12735502 TI - When to biopsy and when to stop biopsying. AB - With the widespread use of serum prostate-specific antigen (PSA) and transrectal ultrasound-guided needle biopsy of the prostate in men with suspected prostate cancer, physicians are faced with the dilemma of treating a patient with a high index of suspicion of prostate cancer but with an initial set of negative biopsies. For the initial biopsy, the optimal number of biopsy cores for detecting prostate cancer in prostate biopsy remains controversial; it is also often unclear who should undergo a repeat prostatebiopsy and when to stop biopsying. PMID- 12735504 TI - Imaging clinically localized prostate cancer. AB - At this time there is no highly sensitive and specific widespread radiographic test for local staging of prostate cancer. Future developments will likely require a combination of imaging modalities with utilization guided by risk stratification models (Table 4). Staging data for all imaging tests discussed in this article are summarized in Tables 5 and 6. Clinically, conventional gray scale TRUS remains the most frequently used tool because of its utility in guiding prostatic biopsies. Modifications of TRUS--including power and color Doppler, 3D imaging, and new ultrasound contrast agents and elastography--show promise in increasing the accuracy of ultrasound. Endorectal MRI may have some value for staging selected patients. The addition of prostatic MRS, which images the differential activity of metabolites, may increase the specificity of MRI. Newer techniques with finer voxel resolution may prove to be clinically useful. A large well-designed study evaluating the utility of MRI/MRS is currently being planned. Cross-sectional imaging of the pelvis with either MRI or CT should be used selectively as should radionuclide bone scans. Similarly, ProstaScint scans should be ordered selectively, either before or after primary therapy, rather than routinely in all patients. PMID- 12735503 TI - Biochemical staging of prostate cancer. AB - PSA continues to be one of the most effective and widely used cancer screening tools available. Its popularity in prostate cancer screening, however, has eroded its usefulness in the staging of this disease. As more men are screened every year on a routine basis with DRE and PSA, the average PSA at diagnosis has drifted down to well below 10 ng/mL in many centers, including ours. This trend is likely to accelerate, as a PSA cut off for prompting biopsy of the prostate of 2.5 ng/mL gains more widespread acceptance. The recent realization that, at these levels, serum PSA is more reflective of the presence of BPH than of the extent of cancer and, therefore, does not provide additional staging information, has renewed the search for new biochemical markers that are capable of predicting prostate cancer stage and prognosis. Because of the heterogeneity of this disease, it is unlikely that a single biochemical marker that is capable of accurately staging all prostate cancer patients will be found. For this reason, nomograms that are capable of integrating various parameters to predict stage and prognosis will remain indispensable. As new biochemical markers that provide independent predictive information about stage or prognosis are identified, they can be incorporated into currently available nomograms. Of the biochemical markers discussed in this article, IL-6sR and TGF-beta1 are the most promising. By incorporating them into a preoperative nomogram designed to predict PSA recurrence, we found that they improved the ability to predict biochemical recurrence by a statistically and clinically significant margin. The ability to stage prostate cancer and predict response to therapy has improved dramatically over the last 3 decades. Nevertheless, there is still a need for new biochemical markers that will improve the ability to predict an individual patient's stage and response to therapy. Incorporating these new markers into nomograms will enhance the ability to provide optimal care for each prostate cancer patient. PMID- 12735505 TI - Emotional and cognitive burden of prostate cancer. AB - The wide-ranging effects of prostate cancer can be an emotional burden to the patient and his family. Recognizing important periods during the diagnosis and treatment that can be particularly stressful, the symptoms indicating patients are at high risk for emotional distress, and the signs and symptoms of emotional distress can encourage improved communication, education, treatment, and referral to minimize the effects of the emotional distress. PMID- 12735506 TI - The physical burden of prostate cancer. AB - The physical burden of prostate cancer is considerable and affects quality of life in men with both localized and metastatic disease. This physical impact results as much from treatment for prostate cancer as from the disease itself. In advanced disease, although patients can experience considerable pain and discomfort from bony lesions, they also can experience bothersome fatigue and sleep disturbances from institution of hormone ablation therapy. In localized disease, although patients can have lower urinary tract symptoms from untreated prostate cancer, all aggressive treatments can result in urinary, sexual, and bowel dysfunction that can bother the patient and affect quality of life. Patients and providers must be vigilantly aware of the physical burden of prostate cancer when initiating treatment for this disease and during follow up after treatment. By being cognizant of the physical impact of prostate cancer on quality of life, providers can address patients' problems early in their course of treatment and maximize patients' HRQOL and overall satisfaction with care. PMID- 12735507 TI - Localized prostate cancer: radiation or surgery? AB - The treatment of localized prostate cancer remains controversial because of the lack of conclusive well-controlled or randomized studies comparing outcomes of radiotherapy to outcomes of radical prostatectomy. A comparison of different therapies should include issues of cancer control, morbidity, quality of life (QOL), salvage of primary treatment failures, late effects, and cost. The available data suggest that these two modalities provide similar rates of cancer control at 10 years, and that except for the youngest patients, choice of therapy should be based on toxicity and QOL issues. PMID- 12735508 TI - Nonmedical factors in treatment selection. AB - Given that no therapy for localized prostate cancer has been proven superior to any other in well-designed clinical trials, the patient and physician may be influenced toward a particular therapy by a number of nonmedical factors. These factors can be logistic, socioeconomic, or related to the patient's perception of the disease and expectations concerning the disease. These factors, along with the absence of a clearly superior therapy, account, at least in part, for the tremendous variance in patterns of care by age, geography, race and education. PMID- 12735509 TI - Mindless or mindful? Radiation oncologists' perspectives on the evolution of prostate cancer treatment. AB - The evolution of radiation therapy treatment for prostate cancer has been striking over the last 10 years. Advances in brachytherapy (BT), external beam radiotherapy (EBRT), and the combination of EBRT + BT have led to improved biochemical and clinical results. This article describes these advances in the context of the treatment decision process. Key to this process is the assignment of patient risk, which is based on the results of conventional radiation dose and techniques. Using the 1992 AJCC palpation staging system, Gleason score, and pretreatment prostate-specific antigen, two different risk assessment algorithms were compared. Both gave comparable approximations of risk, although the single factor high-risk model was superior in differentiating those patients with the highest probability of failing treatment after radiotherapy. Such criteria are the foundation for treatment selection. Objective findings support BT alone or EBRT alone for low-risk patients, high-dose EBRT or EBRT + BT for intermediate risk patients, and EBRT + androgen deprivation for high-risk patients. In summary, advances in radiation oncology have led to significant gains in prostate cancer control. Clinical prognostic factor-based patient selection is central to the optimization of outcome. PMID- 12735510 TI - Practical considerations in permanent brachytherapy for localized adenocarcinoma of the prostate. AB - Prostate brachytherapy has become an accepted treatment modality for localized prostate cancer. Long-term biochemical and biopsy data confirm the early positive impressions that brachytherapy is as valid a treatment option as radical prostatectomy or EBRT. Quality-of-life data also look promising, but more follow up data are needed. Is brachytherapy as good as or perhaps better than radical prostatectomy? This question cannot be answered yet. Well-controlled, randomized studies are needed. In the meantime, the clinician will have to rely on the available published data. PMID- 12735511 TI - Practical considerations in radical retropubic prostatectomy. AB - Overall, in the hands of an experienced surgeon, the outcomes following radical prostatectomy are excellent. Attention to patient selection, preoperative management, surgical technique, and postoperative management are essential factors contributing to favorable outcomes for men with a biologically significant cancer and 10-year life expectancy. For these men, radical prostatectomy represents the optimal management based on cure, morbidity, and quality of life. PMID- 12735512 TI - Complementary and alternative therapies. AB - Patients with prostate cancer increasingly use complementary and alternative therapies. A well-informed oncologist can guide patients to use such treatments to maximize their benefits and reduce the risk of harm. Patients should be dissuaded from using alternative therapies instead of mainstream care to treat cancer. Preliminary evidence suggests that botanical medicines such as PC-SPES may be of benefit, but comparative human trials are yet to be conducted and problems of contamination resolved. Some complementary therapies help reduce symptoms in early-stage prostate cancer, but botanical remedies must be treated with caution because of the possibility of adverse effects and interactions with conventional medicine. PMID- 12735513 TI - Markers and meaning of primary treatment failure. AB - BCR is the most clinically used endpoint for identification of treatment failure. Approximately 15% to 53% of patients undergoing primary curative therapy will develop BCR. BCR often precedes clinically detectable recurrence by years. It does not necessarily translate directly into PCa morbidity and mortality, nor does it always reflect the desired endpoint. Furthermore, it has not been validated as a surrogate endpoint, in that interventions that have been shown to alter the PSA level have not been shown to also alter survival. The utility of PSA level as a surrogate endpoint is brought into question by the knowledge that the overall survival rate of patients at 10 years is similar in patients with and without BCR, and that in a significant proportion of men, the only evidence of disease during their lifetime will be a detectable PSA level. The likelihood of developing BCR post-therapy can be predicted by using multiple clinical and pathologic variables. With the development of nomograms that incorporate several markers, the accuracy of prediction has improved. Until recently, the natural history of BCR post-RRP has not been well understood. Pound et al showed the heterogenous and prolonged natural history of BCR. In this large series of men with BCR following RRP, only 34% of men developed metastatic disease. The median time from development of BCR to identification of metastases was 8 years, and the median time from the development of metastatic disease to death was just under 5 years. These data highlight the extremely variable and potentially indolent nature of BCR. The risk of metastatic disease following BCR has been relatively well defined and relates to PSADT and time to PSA recurrence. It generally is accepted that a PSADT of less than 6 to 10 months and a time to PSA recurrence of less than 1 to 2 years relates to a higher risk of developing metastatic disease. Local recurrence, however, remains poorly understood with respect to its true incidence, clinical significance, and natural history. The significance of BCR post-RT remains unclear due to the lack of data on its natural history. Attempts have been made to identify patients at high risk for metastatic progression by looking at time to PSA recurrence and PSADT. A PSADT of less than 6 to 12 months and a time to PSA recurrence of less than 12 months reflects a higher risk of developing metastatic disease. Accurate risk stratification by means of an algorithm similar to that produced by Pound et al has not been performed on a large cohort, thus making risk assessment for an individual patient difficult. The major dilemma for clinicians in the management of BCR is the identification of the site of disease recurrence, which ultimately guides therapy decisions. Clinicopathologic features allow for risk stratification for recurrence, and multiple investigations have attempted to localize the site of recurrence. Time to biochemical progression, Gleason score, and PSADT are predictive of the probability and time to development of metastatic disease, and allow for stratification of patients into different risk groups (see Table 2). TRUS, CT, PET, and DRE all have limited utility in the identification of local recurrence. ProstaScint and MRI have demonstrated encouraging initial results: however, they require further investigation. Bone scintigraphy is of little value for the initial investigation of BCR. In patients with a PSA level of less than 10 ng/mL, the risk of having a positive bone scan is less than 1% and, until the PSA level rises above 40 ng/mL, the risk of having a positive bone scan is less than 5%. Therefore, bone scintigraphy should be reserved for patients with a PSA level greater than 10 to 20 ng/mL or patients with a rapidly rising PSA level. Using new MRI sequences, there is some evidence that MRI is better for the detection of bony metastatic disease; however, this technique requires further investigation. BCR causes anxiety for the patient and the treating doctor, because the best way to manage patients with PSA-only progression is unknown. Currently, there are no validated treatment recommendations for the management of BCR. The information in this review provides the framework for assignment of patients into clinical trials based on different risk categories. Patients at high risk for metastatic progression could be identified early and thus entered into appropriate clinical trials for systemic therapies. Similarly, patients with a low risk of progression could be placed into observation protocols, potentially sparing them from exhaustive and inappropriate investigations. PMID- 12735514 TI - Selecting a secondary treatment. AB - There is compelling evidence that early hormonal therapy prolongs life in many stages of prostate cancer. Large-scale trials to answer this question have not yet been conducted in surgically treated patients or in patients with PSA-only relapse. Thus, many physicians and patients use early hormone therapy in PSA-only relapse. Many unique new agents are being tested in this population and may offer benefits. Patients and physicians are encouraged to participate in such trials, with hormone therapy reserved for subsequent use. Following failure of primary hormone therapy, a standard algorithm of care exists: antiandrogen withdrawal, use of alternative or first-line anti-androgens. ketoconazole. and chemotherapy. At each interval, clinical trials should be offered since none of these maneuvers are proven to prolong life. PMID- 12735515 TI - Assumed policy similarity and voter preference. AB - The effects of attitude similarity on voters' preferences were examined. Using secondary analyses, the authors created measures of assumed similarity across 6 issues between voters and U.S. presidential candidates (in 1972). Greater similarity was associated with greater attraction (operationalized in terms of voters' presidential preferences). In 2 independent analyses, perceived similarity resulted in predictive accuracy of 84% to 88%. In a 3rd analysis, the predictive efficiency of each of 6 similarity measures was determined and used to develop a model that accurately predicted voters' actions in a hold-out sample. Findings demonstrate the importance of perceived attitude similarity in determining voter preferences and suggest the utility of earlier similarity attraction research for the development of models of policy choice behavior. PMID- 12735517 TI - Attitudes accentuate attributes in social judgment: the combined effects of substance use, depression, and technical incompetence on judgments of professional impairment. AB - Evaluative reactions involving attitudes have been shown to affect social judgments under certain conditions. Researchers assume that personality traits and behaviors toward which people hold strong attitudes evoke evaluative reactions. Such reactions influence the importance of these attributes as they are integrated with other information during the formation of social judgments and, in part, account for the accentuation effect. Social judgment was studied in the context of professional-impairment impressions provided by nurses. Data were modeled using hierarchical techniques to examine the interaction between within judge differences in attribute weighting and between-judges differences in attitudes. Social judgments of fictitious targets were moderated by individual differences in judges' attitudes toward negative target attributes; some, but not all, attributes were accentuated as predicted. PMID- 12735516 TI - Intrafamily aggression in Turkey: frequency, instigation, and acceptance. AB - The present study was designed (a) to determine the frequency of aggression within different pairs of family members, (b) to define actions that may be construed as instigations of intrafamily aggression when committed by different members of the family, and (c) to investigate acceptance rates for different types of aggression from and toward different family members as reactions to different instigations. Questionnaires were administered verbally to 185 Turkish men and women between the ages of 14 and 75. The reported frequency of aggression was highest in mother-child relationships. Reported frequencies were higher for milder acts of aggression than for harsher acts. Content analyses of definitions of different actions that could be construed as instigations for aggression varied, depending on the person performing the act. Variations were consistent with power differences within the family and with gender stereotypes. The acceptance rate for intrafamily aggression was higher for verbal than for physical aggression and showed variations, depending on the nature of the instigation. Discussion focused on the relationship between intrafamily aggression and control. PMID- 12735518 TI - Do attributions change over time when the actor's behavior is hedonically relevant to the perceiver? AB - The tendency for observers to overattribute others' behavior to dispositional variables and, correspondingly, to underestimate the importance of situational variables has been well documented. Yet, the strength of this so-called fundamental attribution error has been challenged by J. M. Burger (1991), who showed that this bias disappeared over time. But does time cause an attribution shift when the other's behavior is negative and hedonically relevant to the observer (e.g., the former is attacked by the latter)? In an exploratory study, the authors showed that even in this case, attributions to dispositional variables tended to diminish overtime. PMID- 12735519 TI - The roles of praise and social comparison information in the experience of pride. AB - The authors examined the roles of social comparisons, publicity of success, and praise on the experience of pride in an experiment in which college students successfully completed a timed intelligence task in private and later received 1 of 4 types of feedback from the experimenter: no feedback (private), mere public acknowledgment of completion, general praise containing both a public and an evaluative component, or praise containing explicit comparison information. Half of the participants also received written normative information suggesting they performed at a high level. Participants then completed a number of dependent measures, including a key measure of pride. Overall, results suggest that the public aspect of a performance, together with the superior standing suggested by any praise accompanying this publicity, is important in the experience of pride. PMID- 12735520 TI - Differences between Saudi and U.S. students in reaction to same- and mixed-sex intimacy shown by others. AB - U.S. and Saudi male and female students were shown 3 photos of 2 women sitting at a close, intermediate, and far distance from each other. Participants were then asked to imagine 2 sisters, 2 brothers, and a brother-sister team sitting at each of these 3 distances and to rate their comfort or discomfort with each scene. As predicted, the U.S. students felt more comfortable the farther away 2 brothers were sitting and the closer a brother and sister were sitting, whereas the Saudi students showed the opposite reaction. It was suggested that distances between the sexes in conservative Muslim countries is maintained even in intrafamily contact and that the aversion to public displays of male-male intimacy found in the United States and Europe does not generalize to other cultures. PMID- 12735521 TI - The gendered nature of family structure and group-based anti-egalitarianism: a cross-national analysis. AB - Using 4 samples of adolescents from 3 nations (Australia, Sweden, and the United States), the authors explored whether the gendered nature of the family socialization environment affected young people's level of group-based social egalitarianism. It was hypothesized that the greater the father's influence in the family, the greater the children's level of group-based social anti egalitarianism. The results were consistent with the authors' expectations. Children from father-headed households had the highest level of group-based social anti-egalitarianism; children from mother-headed households had the lowest level of group-based anti-egalitarianism; and children from dual-parent households were in between. Similarly, children from homes in which the father had the greatest decision-making power tended to exhibit the highest levels of anti-egalitarianism, whereas children from homes in which the mother had the greatest decision-making power displayed the lowest levels of social anti egalitarianism. Family structure did not interact with either the nationality or gender of the child. PMID- 12735522 TI - Attributional style predicts causes of negative life events on the Attributional Style Questionnaire. AB - The causes cited by 218 participants for the hypothetical positive and negative life events on the Attributional Style Questionnaire (ASQ) and the dimensional ratings of the causes were examined to determine the match between the dimensional and categorical definitions of attributional style. Optimists (n = 105) and pessimists (n = 113) used different types of causes to explain the negative ASQ events, but not the positive events. However, optimists' and pessimists' causal explanations shared a number of features. The findings suggest that attributional styles depend, in part, on the event being explained and demonstrate that the ASQ events elicit specific types of causes. PMID- 12735523 TI - Cultural meanings of nature: an analysis of contemporary motion pictures. AB - To evaluate current cultural meanings of nature, the authors asked 65 undergraduate students to "list 3 movies in which nature was an important aspect of the film." They also were asked to specify the natural element that stood out to them and describe "how it related to the overall theme of the movie." Two independent groups of raters skilled in interpretive analysis developed thematic meanings from these responses. Following this, in a 2nd study, a different group of participants rated the 18 most frequently mentioned natural elements on thematic scales derived from the initial interpretive analysis. Participants in the 1st study mentioned 33 different movies at least twice and 5 themes that captured the meaning of nature in these films. Correlational results derived from the 2nd study indicated that rating scales reflecting these 5 themes formed 2 distinct groups; the first group described settings in which nature is experienced as adversarial and plays a significant role in dramatic action, and the second group defined settings in which nature is viewed either as a place of refuge or simply as a locale in which ongoing narrative action occurs. The general conclusion reached in both studies concerns the often noted but not always appreciated fact that movies--similar to everyday events and actions- always take place in specific settings and that neither life events nor dramatic narratives can be understood apart from specific settings. PMID- 12735524 TI - The difference between playing games with and without the computer: a preliminary view. AB - The authors address the question of whether associations between video games and cognitive and metacognitive variables depend either on the features of the computer or on the content of the game that the computer allows one to play. An experiment to separate these two kinds of effects was carried out by using a traditional version and a computer-supported version of Pegopolis, a solitaire game. The two versions were exactly the same except that they were played by moving pieces either on a real board or on a virtual computer-presented board. The performance levels and strategies followed during the game by the 40 undergraduates who took part in the experiment were not significantly different in the real and virtual conditions. None of the participants transferred playing strategies or practice from one version of the game to the other. Scores were not affected by gender or by the studies pursued by participants, the habit of playing games in the traditional manner or playing video games, or intelligence. Retrospective reports did not support differences in the subjective experience between the two versions. Results showed that video games, when they do not make much use of the computer's special features, produce effects because of the situations they simulate rather than because of features of the computer itself. PMID- 12735525 TI - Behavioral and personality characteristics of children with reactive attachment disorder. AB - The authors compared behavioral and personality characteristics of children with reactive attachment disorder (RAD) with non-RAD children. Participants included parents of children with RAD (n = 21), parents of non-RAD children (n = 21), and some of the children (n = 20). The parents completed questionnaires regarding behavioral and personality characteristics of their children. Parents were also given the option of asking their children to participate in the study by completing self-report measures. Several significant findings were obtained. Children with RAD scored lower on empathy but higher on self-monitoring than non RAD children. These differences were especially pronounced based on parent ratings and suggest that children with RAD may systematically report their personality traits in overly positive ways. Their scores also indicated considerably more behavioral problems than scores of the control children. Previous research has been generally qualitative in nature. The current research represents some of the first quantitative, empirical work documenting specific behaviors associated with the diagnosis of RAD. The findings of this study have implications for better understanding and dealing with reactive attachment disorder. PMID- 12735526 TI - The assessment of attitudes toward individuals with disabilities in the workplace. AB - The authors conducted 2 studies to develop and test measures that assess beliefs about what constitutes a disability, affective reactions to working with individuals with disabilities, and beliefs about the reasonableness of workplace accommodations, in general and within the context of the Americans With Disabilities Act (ADA). The results of these 2 studies showed substantial differences in what was considered to be a disability. In general, more physical and sensory-motor conditions were considered disabilities than were psychological conditions. Furthermore, the conditions believed to be disabilities did not necessarily match what is covered by the ADA. Gender and experience with individuals who are disabled were also found to predict affective reactions and the reasonableness of accommodations. Implications for organizations are discussed. PMID- 12735527 TI - The role of peers and families in predicting the loneliness level of adolescents. AB - The authors investigated the relative contribution of peer relations, family structure, and demographic variables in predicting loneliness in adolescents. Ninth-grade high school students (N = 756) from 8 different schools representing various socioeconomic status in Ankara, Turkey, completed the UCLA Loneliness Scale (D. Russell, L. A. Peplau, & M. L. Ferguson, 1978), the Family Structure Assessing Instrument (A. Gulerce, 1996), and an author-constructed questionnaire involving demographic information and variables on peer relations. Results of multiple regression analyses indicated that all three sets of variables accounted for 41% of the variance in loneliness scores. Additionally, peer relations contributed 34% of the variance, family structure 14%, and demographic variables 3%. Within the limits of the study, peer relations appear to be the best predictors of adolescent loneliness. PMID- 12735528 TI - Is cognitive therapy consistent with what we know about emotions? AB - The cognitive perspective has been a dominant force in psychology and psychotherapy since the 1970s. Cognitive therapies share an emphasis on the priority of changing cognition as the key to bringing about changes in patterns of dysfunctional emotional reactions and symptomatic behaviors. In this article, the author examined the basis for the assumptions of cognitive therapy in the context of recent evidence of the long-term effects of stressful experiences. This evidence contradicts the assumption of many cognitive therapists that changing cognition is prior to and necessary to changing emotional reactions. Neurobiological evidence indicates that emotions can be experienced without cortical interpretations of stimuli, and clinical evidence indicates that experiences can be stored as isolated affective fragments that function later to distort cognition. This suggests that cognitive therapies are based on a limited model of mental functions that sometimes must be supplemented by broader psychodynamic concepts if lasting changes are to occur. PMID- 12735529 TI - Assessing qualitative research in rehabilitation: guidelines from the society for research in rehabilitation. AB - There are numerous contested issues in qualitative research, and its findings are difficult to condense. Thus, short, qualitative research abstracts are difficult both to compile and evaluate. The Society for Research in Rehabilitation has produced guidelines that aim to illuminate and inform the development and judging of such abstracts. These draw on an extensive Health Technology Assessment review of qualitative methods in health services research. PMID- 12735530 TI - The needs and experiences of caregivers of individuals with multiple sclerosis: a systematic review. AB - PRIMARY OBJECTIVE: To appraise recent studies regarding the needs and experiences of caregivers of individuals with multiple sclerosis (MS). DESIGN: The following computerized databases were searched: CINAHL, BIDS IBSS, ASSIA, MEDLINE, PSYCHINFO, British Nursing Index, ISI Web of Science, Zetoc, AMED (1990-April 2002). The computer-based search was supplemented by manual searches of the reference lists of all retrieved studies and review articles. Inclusion and exclusion criteria were formulated. RESULTS: Twenty-four studies from across the world that met the inclusion criteria were reviewed. The majority of studies were descriptive in nature. The studies covered a variety of topics, including how carers assist people with MS, the effect of providing care on a carer's physical and psychological well-being, social life, financial situation and overall quality of life, and how carers cope with the stresses of providing care. CONCLUSIONS: Providing care for a person with MS has a major impact on all areas of the caregiver's life. Perceived social support has been shown to have a beneficial impact on the caregiver. Limitations in design and variation in methodology of studies limits the generalizability of findings. There is a need for further research, in particular the development of reliable and valid disease specific caregiver assessment instruments. PMID- 12735531 TI - A study of interventions and related outcomes in a randomized controlled trial of occupational therapy and leisure therapy for community stroke patients. AB - OBJECTIVE: To undertake a detailed analysis of therapy provided in a multicentred randomized controlled trial of activities of daily living (ADL) and leisure (TOTAL), testing the hypothesis that specific interventions given in the trial affected specific aspects of outcome. SUBJECTS: Three hundred and nine stroke patients who had been randomly allocated to receive either occupational therapy aimed at ADL activities (n = 156) or leisure (n = 153). MEASURES: Number, duration and type of activity undertaken per patient. Barthel Index, Extended Activities of Daily Living Scale (EADL) and Nottingham Leisure Questionnaire (NLQ) six months after entry to the study. METHOD: Activities that had been used in treatment were coded and categorized. Frequently used activities identified. These activities were matched to items from the six-month outcome measures. Patient independence in these outcome items was compared between the leisure and ADL groups. RESULTS: Three hundred and nine therapy record forms were returned. Patients received a median of ten sessions with a median duration of 55 minutes. The ADL group received significantly more, mobility training, transfer training, cleaning, dressing, cooking and bathing training (chi-squared, p < 0.05). Sport, creative activities, games, hobbies, gardening, entertainment and shopping were used significantly more in the leisure group (chi-squared, p < 0.05) than the ADL group. Fifteen items from the outcome measures were identified as specific to these interventions. There were no statistically significant differences in outcome on these 15 items between the ADL and leisure groups (chi-squared, p > 0.05). CONCLUSIONS: We found no evidence that specific ADL or leisure interventions led to improvements in specific relevant outcomes. We believe that these findings should prompt a review of the relationship between process and outcome of occupational therapy. PMID- 12735532 TI - Use of videotape to assess mobility in a controlled randomized crossover trial of physiotherapy in chronic multiple sclerosis. AB - OBJECTIVES: To determine to what degree assessment of mobility based on comparison of videotape recordings before and after courses of physiotherapy in patients with chronic multiple sclerosis (MS) is reliable, correlates with 'live' assessments and indicates benefit. DESIGN: Prospective data collection within a randomized crossover controlled trial of physiotherapy at home, as an outpatient, or 'no therapy' in 40 patients. SETTING: Hospital outpatients: outpatient and home physiotherapy. OUTCOMES: Mobility change based on a comparison of short video recordings before and after each treatment period was scored independently by two physiotherapists blinded to therapy type and other measures of outcome. Scores were compared with changes in the Rivermead Mobility Index (RMI) and other indices assessed by a physiotherapist in the patient's home. RESULTS: The two video observers agreed substantially on patient outcome. Changes in walking based on video correlated with RMI for home treatment (r = 0.41, p = 0.008) but not for hospital or no treatment periods (r = 0.14 and 0.15): video changes correlated with the 'live' assessor's global change score inconsistently ('no therapy' r = 0.48, p = 0.002, hospital r = 0.30, p = 0.06 and home r = 0.17, p = 0.30 treatment periods). Based on video data alone, improved mobility was evident following home therapy for only one observer but not for the other or the averaged scores of both. CONCLUSION: There was substantial agreement between two observers deciding on change in mobility based on independent blinded evaluation of short video sequences. However the correlations of these with 'live' assessments were variable. Physiotherapy had a less clear benefit on mobility based on video analysis alone compared with 'live' assessments. The study highlights the need for more objective measures of habitual mobility over longer periods. PMID- 12735533 TI - Agreement between physiotherapists on quality of movement rated via videotape. AB - BACKGROUND: Although achieving quality of movement after stroke is an important aim of physiotherapy it is rarely measured objectively or described explicitly. OBJECTIVE: To test whether physiotherapists agree on a composite measure of quality of movement. SETTING; A movement analysis laboratory SUBJECTS: Ten stroke patients and 10 healthy age-matched volunteers. DESIGN: Prospective correlational. PROCEDURE: All subjects were videofilmed performing three trials of six standardized functional tasks. Two videotapes were made, each with a different randomized order of appearance of the trials. Ten senior physiotherapists independently rated the videotapes twice using a 100-mm visual analogue scale. ANALYSIS: Analysis of variance models were fitted to transformed data. Estimates of components of variance were calculated and presented as a percentage of the total variance for differences, within subjects (intra subject), between raters (inter-rater) and within raters (intra-rater). An acceptable percentage was set at less than 10%. RESULTS: The percentage of intra subject variance ranged from 1% (pick up box and walking) to 9% (step on block). The percentage of inter-rater variance ranged from 18% (pick up pencil) to 38% (sit to stand). The percentage of intra-rater variance was less than 1% for all tasks. CONCLUSIONS: Although physiotherapists disagreed with each other on quality of movement they were more consistent in their own scoring. PMID- 12735534 TI - Psychometric properties of the Rivermead Mobility Index in Italian stroke rehabilitation inpatients. AB - OBJECTIVE: To examine the internal consistency, validity, responsiveness and test scalability of the Rivermead Mobility Index (RMI) in Italian rehabilitation inpatients recovering from stroke. DESIGN: Seventy-three stroke inpatients undergoing rehabilitation were assessed at admission (T0) and five weeks later (T1), using RMI, the motor (motFIM) and cognitive (cognFIM) subscales of the Functional Independence Measure, the 'leg' section of the Motricity Index and Trunk Control Test. RESULTS: Cronbach's alpha of the RMI was 0.92. The item-to total correlation coefficients (r(rb)) ranged from 0.36 to 0.83, all p < 0.003. All correlations between RMI scores and the other instruments, both at T0 and T1, were statistically significant (r > or = 0.49, all p < 0.0001), except those with cognFIM. The difference in RMI scores over the testing period was statistically significant (sign test: z = 7.1, p < 0.0001) and the effect size was 0.89. The coefficient of reproducibility was 0.95 at T0 and 0.93 at T1, and both coefficients of scalability were 0.67. CONCLUSIONS: The study confirms the internal consistency, construct validity and responsiveness of the RMI, according to the classic psychometric indexes. However, some minor concerns arise regarding: (a) a floor effect of RMI in subacute rehabilitation stroke inpatients at admission and; (b) one item ('bathing') that seems sensitive to cultural and environmental factors. Moreover, even though RMI met the scaling criteria, the item hierarchy is not coincident with the one originally postulated. So, RMI should be considered only as a summated index with ordinal properties, and not a hierarchically ranked scale. PMID- 12735535 TI - The validation of the Subjective Index of Physical and Social Outcome (SIPSO). AB - OBJECTIVE: To validate the Subjective Index of Physical and Social Outcome (SIPSO) in its final 10-item version as a measure of reintegration to pre-stroke lifestyle. DESIGN: Postal and interview administration of questionnaires. MEASURES: SIPSO, Functional Limitations Profile (FLP), Reintegration to Normal Living Index (RNLI). SETTING: Community setting, Bath, UK. SUBJECTS: Two hundred and sixty-one survivors of stroke discharged from a district general hospital. RESULTS: Internal consistency (item-total correlations >0.6) and test-retest reliability (intraclass correlation coefficient (ICC) = 0.96) of the measure were confirmed. Construct validity was confirmed through significant correlations with the six subsections of the FLP and the RNLI. Confirmatory factor analysis of the measure found that the first five items form a robust subscale that clearly relates to physical function. The items within the second subscale (questions 6 10) measure several domains (social, leisure, self-image) and do not form a homogeneous group. The SIPSO was able to detect an improvement in integration within a sample of patients over a three-month period soon after discharge (effect size = 0.26). The hypothesis that the degree of improvement experienced by patients following discharge from hospital would be small, but should be detectable was confirmed. The agreement between proxy and patient on the SIPSO was acceptable for individual items (kappa value range = 3.1-5.8) and total score (ICC = 0.96). CONCLUSIONS: The SIPSO is a brief self-complete measure, with proven validity and reliability, which addresses both quantitative and qualitative aspects of activities and interaction. It provides a useful measure for evaluating rehabilitation programmes that aim to reintegrate patients to their pre-stroke lifestyle. The authors suggest that the SIPSO should be used as a 10-item measure. PMID- 12735536 TI - A biomechanical investigation into the validity of the modified Ashworth Scale as a measure of elbow spasticity. AB - OBJECTIVE: To investigate the criterion validity of the modified Ashworth Scale. POPULATION: Volunteers from a stroke population admitted to a district general hospital stroke unit diagnosed with a first ever stroke less than 26 weeks previously. OUTCOME MEASURES: Resistance to passive movement about the elbow was simultaneously quantified (biomechanically) and graded (modified Ashworth Scale). Passive range of movement and peak instantaneous velocity during passive movement were also measured. ANALYSIS: Criterion validity was investigated as convergent construct validity (using the Spearman's correlation coefficient) and concurrent validity (using analysis of variance). RESULTS: One hundred measurements were taken on 63 subjects. Correlation between the modified Ashworth Scale and resistance to passive movement was 0.511. Resistance to passive movement and velocity showed significant differences between the modified Ashworth score of '0' and a modified Ashworth score greater than '0' (p < 0.01). There were no significant differences between MAS '1', '1+' and '2'. Resistance to passive movement in the impaired arm was significantly higher than in the nonimpaired arm (p < 0.01). CONCLUSION: The modified Ashworth Scale does not provide a valid measure of spasticity at lower grades but it may provide a measure of resistance to passive movement. PMID- 12735537 TI - Multiple sclerosis in Stockholm County. A pilot study exploring the feasibility of assessment of impairment, disability and handicap by home visits. AB - OBJECTIVE: A pilot study performed within Stockholm County to evaluate the feasibility of collecting data using a comprehensive evaluation package administered in the home environment to assess impairment, disability and handicap in order to explore the consequences of multiple sclerosis (MS). DESIGN: Home visits to 26 purposefully selected MS patients with different levels of disability, in both ordinary and sheltered living. The comprehensive evaluation package included: biographical data, Mini-Mental State Examination, Free Recall and Recognition of 12 Random Words Test, Symbol Digit Modalities Test, Beck Depression Index, Lindmark Motor Capacity Assessment, time to walk 10 metres, Nine-hole Peg Test, Barthel ADL Index, Katz Extended ADL Index, Frenchay Activities Index, Sickness Impact Profile and frequency of falls and injurious falls. RESULTS: This pilot study demonstrates that the proposed methods can be used to evaluate MS patients differing in levels of disability and forms of living. The data collection method, based on home visits, was well accepted by the patients, their spouses and salaried personal assistants and could be performed within 2-2 1/2 hours. CONCLUSIONS: The evaluation package used in this pilot study is suitable for use in population-based studies and it should provide comprehensive information on the impact and consequences of MS on patients, and should contribute to the identification of areas in which the provision of rehabilitation and health care services needs to be improved. PMID- 12735538 TI - Shoulder pain in people with a stroke: a population-based study. AB - OBJECTIVE: To measure the occurrence of shoulder pain after stroke. To identify the factors that predict risk of shoulder pain after stroke. DESIGN: Auckland Stroke Study, population-based case-cohort study. SETTING AND SUBJECTS: All cases of stroke, including those managed outside hospital, over a 12-month period ending February 1992 were considered in Auckland. OUTCOME MEASURES: Self-reported shoulder pain at one week, one month and six months after the onset of stroke for each person. RESULTS: A total of 1,761 stroke events were identified. Self reported shoulder pain among survivors increased from 256/1474 (17%) at one week, to 261/1,336 (20%) at one month and 284/1,201 (23%) at six months. Shoulder pain was positively associated with motor deficit, side of deficit and severity of deficit. In those surviving to six months after stroke, the risk of shoulder pain was higher in those with severe upper limb motor deficit (odds ratio (OR) 4.94; 95% confidence interval (CI) 3.06-7.98) and in diabetics (OR 1.57, 95% CI 1.15 2.14). Risk of shoulder pain increased with time and was lower for those in institutional care. CONCLUSION: Shoulder pain after stroke is common, especially in patients with severe sensorimotor deficits, diabetics and those living at home. Appropriate management may reduce the rate of occurrence. PMID- 12735539 TI - Caregiver strain in spouses of stroke patients. AB - OBJECTIVE: To test the ability of a previously generated logistic regression model to predict caregiver strain from carer mood, negative affectivity and perceived patient functional ability. DESIGN: Postal prospective survey. SETTING: Spouses of community-residing patients identified from hospital stroke registers. METHOD: Spouses were assessed at three and six months after stroke. A previously derived equation was used to make predictions at three months of their level of strain at six months, which were compared with observed outcomes. MEASURES: Spouses were asked to complete the Caregiver Strain Index (CSI), the General Health Questionnaire (GHQ-12), the Positive and Negative Affectivity Schedule (PANAS) and to assess patients' independence in activities of daily living on the Extended Activities of Daily Living Scale (EADL). RESULTS: Of 409 stroke patients, 276 had an identifiable co-resident spouse and 116 (42%) completed the measures. At three months after stroke, 39 carers (34%) were under significant strain with 40 (35%) under strain at six months. The predictive model using the GHQ-12, PANAS and EADL at three months was 78% accurate in predicting levels of caregiver strain at six months. CONCLUSION: Carers at risk of later strain could be identified for further follow-up. Services to provide emotional support to carers might be effective in the reduction of carer strain. PMID- 12735540 TI - An exploratory study of the consistency of balance control and the mobility of people with Parkinson's disease (PD) between medication doses. AB - OBJECTIVE: To evaluate the consistency of balance control, mobility and perceived difficulties with functional tasks by people on carefully controlled anti parkinsonian medication programmes, between doses on a single day and across days using standard clinical measures. DESIGN: Small group prospective study. SETTING: A hospital outpatient physiotherapy department. SUBJECTS: Subjects, recruited from a specialist Parkinson's disease (PD) clinic, had to be independently mobile, diagnosed with PD at least five years previously and on a stable drug regime. METHOD: Assessments (timed up and go, functional reach and a tap test) were completed 30 minutes prior to medication and hourly for 4 hours. They were repeated on a second day within a seven-day period. The self-assessment of functional disability was completed during the worst and best periods of the drug cycle. A general linear model (repeated measures analysis of variance) was used to analyse the data. RESULTS: Nine subjects (8 males) with PD (grade III Hoehn and Yahr, average duration of symptoms eight years), average age 72 years, were recruited to the study. Four subjects had worse Hoehn and Yahr scores (grade 4) during the worst phase of the L-dopa cycle. Balance and mobility data did not change significantly over the medication cycle. Tests on day 2 were similar to day 1 with only the tap test showing a significant mean difference (p = 0.02) (2.6, 95% confidence interval (CI) 0.5-4.8). Individuals perceived significantly greater difficulty (mean difference = 7.9, 95% CI 3.6-12.2) (p < 0.001) in achieving functional tasks at the end of dose than peak dose of the medication cycle. CONCLUSION: Measures of balance and mobility remained stable over the cycle. Fluctuations in the perception of difficulties with daily functional tasks were not accompanied by observed fluctuations in performance. The improvement in the mean of the tap test over the days could reflect a learning effect. PMID- 12735541 TI - Is stair negotiation measured appropriately in functional assessment scales? AB - BACKGROUND: A decline in mobility may result in problems with the negotiation of stairs, which can potentially be hazardous. In practice, stair negotiation is an important aspect of daily living and therefore needs to be assessed carefully. METHODS: We conducted a systematic literature review to identify the way functional assessment scales assess stair negotiation. We evaluated whether stair negotiation could be assessed in a valid and reliable way with these scales. RESULTS: Forty-three of the 92 identified scales have an item on stair negotiation. In these scales, the definition of 'negotiation of stairs' varies, as does the definition of independence. Important aspects such as safety on stairs are missing from all scales. In contrast to older scales, newer scales consist of items that have been tested for validity and reliability. In none of the scales was the stair negotiation item tested separately for validity. Only two scales examined test-retest reliability and only one measured inter-observer agreement. DISCUSSION: In current functional assessment scales stair negotiation is measured with great heterogeneity and insufficient validity. In patients and in studies in which assessment of stair negotiation ability is a key part of functional assessment, an improved, well-validated scale is needed. This scale should include not only the subject's physical ability to negotiate stairs, but also safety and change in ability over time. PMID- 12735542 TI - Falls risk following discharge from a geriatric day hospital. AB - OBJECTIVE: To assess the predictive validity of the Elderly Mobility Scale (EMS), Functional Reach (FR) and the Barthel Index (BI) in identifying recurrent fallers following discharge from a geriatric day hospital (GDH). SUBJECTS: Seventy-six GDH patients with identified mobility problems. METHODS: Each patient was assessed by an independent physiotherapist before and after a programme of physiotherapy using the EMS, FR and BI and a 'falls follow-up' assessment was performed after four months. RESULTS: Fifty-four per cent (29 out of 54) of patients had reduced mobility at follow-up. Twenty-five per cent (18 out of 71) of patients had two or more falls post discharge. Using logistic regression analysis for EMS, FR and BI at discharge, each of these variables individually was significantly associated with the risk of having two or more falls (p = 0.008, 0.017 and 0.031 respectively). A prognostic tree was developed for GDH patients identifying high-risk and low-risk groups. CONCLUSION: EMS, FR and BI were all significantly associated with GDH patients with mobility problems having two or more falls. A prognostic tree identified high- and low-risk groups of GDH patients and should now be tested prospectively. PMID- 12735543 TI - Co-ordination of patient care in inpatient rehabilitation. AB - OBJECTIVE: To evaluate the effectiveness of a new patient care co-ordinator position in an inpatient rehabilitation setting. DESIGN: Pre and post test using a 42-item staff survey. SETTING: One inpatient rehabilitation service in rural New South Wales, Australia. SUBJECTS: Members of the multidisciplinary team. INTERVENTION: A new patient care co-ordinator position. OUTCOME MEASURES: Comparison of trends in staff responses to pre and post test surveys. FINDINGS: The post test survey demonstrated an overall increase in the reported frequency of desirable rehabilitation practices and increased staff satisfaction with those practices. CONCLUSIONS: A patient care co-ordinator may contribute to improving the processes and practices of inpatient rehabilitation and staff satisfaction. PMID- 12735544 TI - A study of family support, friendship, and psychological well-being among older women in Hong Kong. AB - The "dual-channel" hypothesis (Lawton, 1996), which suggests the dual-antecedent pattern for positive and negative aspects of psychological well-being, was tested by examining the differential relationships between objective and subjective measures of family support (family contact, family quality, perceived importance of family) and friendship (friends support, friends quality, perceived importance of friendship) to two facets of psychological well-being (positive and negative affect). Face-to-face interviews were conducted with a sample of 60 older women aged 60 to 85 in one district of Hong Kong. A series of hierarchical multiple regression analyses, controlling for age and marital status, demonstrated that two subjective measures (family quality and perceived importance of friendship) were significant predictors of positive affect; and one subjective measure (family quality) was a significant predictor of negative affect. The "dual channer" hypothesis was partially supported. Recommendations regarding informal support provision for older women are discussed. PMID- 12735545 TI - Cognitive representations of future gains, maintenance, and losses in the second half of life. AB - The aim of this study is to explore three different developmental dimensions in an aging population. Based on sentence completion responses, the investigation examines personal anticipations of possible gains, maintenance, and losses. Additionally, the effects of age and other personal and situational factors are examined. The study sample consists of 2,934 participants ranging from 40 to 85 years of age, who participated in the German Aging Survey of 1996. Study findings indicate that, to a large extent, the anticipated gains include positive changes in the way of life and increased leisure projects while anticipated maintenance refer to physical and behavioral resources and to life style. Anticipated losses are related to concerns about external living conditions and physical decline. There is a strong association of anticipated gains and maintenance with age, while present health conditions are related to expectations of loss. The implications of the study results for lifespan expectations in the second half of life and for lifespan theory are discussed. PMID- 12735546 TI - The effects of priming on children's attitudes toward older individuals. AB - The purpose of the present research was to examine younger (7-years-old) and older (10-years-old) children's attitudes toward older individuals following one type of five primes: positive prime, negative prime, elderly prime, grandparent prime, or neutral prime. Overall, children's attitudes on three tests- Apperception, Semantic Differential, and Attribute Salience--were affected by the type of prime children were given, with positive and grandparent primes resulting in more positive views toward older individuals than negative, elderly, or neutral (control group) primes. The present research provides evidence that priming the most accessible cognitions about an individual can affect even young children's perception of the individual. These results are discussed in terms of category-based and data-driven processing and may explain the disparate findings obtained in previous studies that have shown the children's attitudes toward older individuals are sometimes negative, whereas other studies have shown that children's attitudes are more positive or neutral. PMID- 12735547 TI - Elder respect among American college students: exploration of behavioral forms. AB - This study explored the ways in which college students conveyed elder respect. Based on data from a questionnaire survey of 521 subjects, 12 forms of elder respect, ranging from care respect to celebratory respect, were measured. Of these forms, six--acquiescent respect, care respect, linguistic respect, salutatory respect, consulting respect, and presentational respect--were frequently practiced. This study provides insights as to how younger adults in American culture exhibit elder respect. A typology of the forms signifying elder respect among the young American adults is presented. Implications of the findings and suggestions for future research are discussed. PMID- 12735548 TI - Presidential address. A partnership in courage. PMID- 12735549 TI - Simulation models to predict outcome after aortic valve replacement. PMID- 12735550 TI - Cognitive changes with coronary artery disease: a prospective study of coronary artery bypass graft patients and nonsurgical controls. AB - BACKGROUND: Cognitive impairment after coronary artery bypass grafting (CABG) is well recognized, but previous investigations have been limited by lack of an appropriate control group. We compared changes in cognitive performance at 3 and 12 months after CABG with those in a control group of patients with comparable risk factors for coronary artery disease (CAD) who had not undergone surgery. METHODS: Patients undergoing CABG (n = 140) and demographically similar nonsurgical control subjects with CAD (n = 92) completed baseline neuropsychological assessment and were followed prospectively at 3 and 12 months. Cognitive function was evaluated with a battery of neuropsychological tests assessing the cognitive domains of attention, language, verbal and visual memory, visuoconstruction, executive function, and psychomotor and motor speed. RESULTS: The CABG patients who were tested in their hospital rooms before surgery had lower scores for timed tests; however, after adjustment for demographic variables and testing location there were no statistically significant differences between the CABG and nonsurgical control subjects in baseline neuropsychological test performance. Both groups improved from baseline to 3 months; the only statistically significant group difference was a greater improvement in the CABG group with regard to verbal memory. At 12 months there were no statistically significant differences between the two groups. CONCLUSIONS: The prospective longitudinal neuropsychological performance of patients with CABG did not differ from that of comparable nonsurgical control subjects with CAD at 3 months or 1 year after base line examination. This suggests that the previously reported cognitive decline during the early postoperative period after CABG is transient and reversible. Continued follow-up will determine whether a specific "late decline" occurs in CABG patients but not in nonsurgical control subjects with similar risk factors for cardiovascular and cerebrovascular disease. PMID- 12735551 TI - Impact of single clamp versus double clamp technique on neurologic outcome. AB - BACKGROUND: Atherosclerotic disease of the aorta has been identified as a risk factor for neurologic complications following coronary artery bypass grafting (CABG) due to the use of aortic clamping and manipulation. We reviewed a change from double clamp to single clamp technique to determine its impact on neurologic outcomes. METHODS: Patients undergoing isolated CABG by a single surgeon were identified as having double clamp technique (DCT) (aortic cross clamp + sidebiting clamp) or single clamp technique (SCT) (aortic cross clamp only). Data were collected by study personnel and clinicians to determine stroke and neurologic injury (confusion, delirium, seizure, altered mental status, and agitation) outcomes for 461 patients. RESULTS: Two hundred seventy-two patients had DCT and 189 patients had SCT performed. There were no differences in mean age, previous stroke, hypertension, or diabetes. Intraoperatively, patients with SCT had shorter bypass times (115 minutes vs 128 minutes, p = 0.001), longer aortic cross clamp time (89 minutes vs 80 minutes, p = 0.001), fewer coronary grafts (2.8 vs 3.1, p = 0.001), and had higher mean arterial blood pressure on cardiopulmonary bypass (76 mm Hg vs 69 mm Hg, p = 0.001). Postoperatively, the SCT group had fewer strokes (1.1% vs 2.9%, NS), and neurologic injuries (3.2% vs 9.6%, p = 0.008). By multivariate analysis, the factors that were related to neurologic injury were DCT (p = 0.04), age (p = 0.001), and number of coronary grafts (p = 0.03). CONCLUSIONS: This experience suggests that the use of the SCT may be important in reducing neurologic injury following CABG. PMID- 12735552 TI - Outcomes and perioperative hyperglycemia in patients with or without diabetes mellitus undergoing coronary artery bypass grafting. AB - BACKGROUND: The association between perioperative hyperglycemia and outcomes in patients with and without diabetes mellitus undergoing coronary artery bypass grafting is not well defined. We measured the association between perioperative hyperglycemia and outcomes among patients undergoing coronary artery bypass grafting. METHODS: We report a historic cohort study of 1574 patients who had undergone coronary artery bypass grafting between 1998 and 1999, 545 (34.6%) with diabetes. Perioperative blood glucose level was defined as the average of all blood glucose tests obtained on the day of and the day after surgery. Outcomes were 30-day mortality, infection rates (sternum, harvest site, sepsis, pneumonia, urinary tract), and resource utilization. RESULTS: After adjusting for diabetes status and calculated preoperative mortality or mediastinitis risk scores, each 50 mg/dL (2.78 mmol/L) blood glucose increase was not statistically associated with higher mortality (odds ratio 1.37; 95% confidence interval, 0.98 to 1.92; p = 0.07), or higher infection rate (odds ratio 1.23, 95% confidence interval 0.94 to 1.60; p = 0.14). Each 50 mg/dL blood glucose increase was associated with longer postoperative days by 0.76 days (95% confidence interval 0.36 to 1.17 days; p < 0.001), increased hospitalization charges by 2824 dollars (95% confidence interval 1599 dollars to 4049 dollars; p < 0.001), and increased hospitalization cost by 1769 dollars (95% confidence interval 928 dollars to 2610 dollars; p < 0.001). In the unadjusted analysis, infections occurred more frequently in patients with diabetes (6.6% vs 4.1%, p = 0.03). CONCLUSIONS: Perioperative hyperglycemia is associated with increased resource utilization in patients undergoing coronary artery bypass grafting with and without diabetes. PMID- 12735553 TI - Emergency surgery after unsuccessful coronary angioplasty: a review of 15 years' experience. AB - BACKGROUND: Emergency coronary artery bypass grafting (CABG) is occasionally necessary for failed percutaneous transluminal coronary angioplasty (PTCA). The aim of this study was to assess the outcome of patients receiving emergency CABG after unsuccessful PTCA over a 15-year study period. METHODS: From January 1982 through December 1996, 74 patients underwent emergency CABG after unsuccessful PTCA (crash group). This group was compared with a matched group of 74 patients having primary elective CABG (control group). RESULTS: All 74 crash group patients were to have PTCA of one coronary system. After PTCA failure, 58 patients (78.3%) developed electrocardiographic changes of evolving acute myocardial infarction (AMI). The overall rate of AMI was 8.1% for the crash group and 2.7% for the control group. Two patients in the crash group died, with no deaths in the control group. There was no significant difference between mean in hospital length of stay. CONCLUSIONS: With prompt, aggressive, and complete myocardial revascularization, patients who required emergency CABG after PTCA failure had an outcome not significantly different from that of patients having elective CABG. PMID- 12735554 TI - Determinants of mid- and long-term results in patients after surgical revascularization for ischemic cardiomyopathy. AB - BACKGROUND: The revascularization of patients suffering from ischemic cardiomyopathy is possible with acceptable perioperative mortality and morbidity. Many publications have discussed the problem of predicting myocardial viability, whereas the quality of the peripheral coronary vessels has been focused on less frequently. METHODS: We studied 908 consecutive patients with ischemic cardiomyopathy revascularized between January 1, 1988 and April 30, 2000. Death, recurrent heart failure, hospitalization due to cardiac causes, ventricular assist device implantation, heart transplantation, and ventricular arrhythmias were defined as adverse events. To analyze the importance of pre- and perioperative variables (state of the coronary arteries, myocardial viability, complete vs incomplete revascularization, urgency of the operation, previous operations, gender, diabetes, preoperative New York Heart Association class, age, number of grafts, and ischemic time), a proportional hazards model was used. RESULTS: The most important predictors of short- and long-term event-free survival were the quality of the coronary arteries, followed by myocardial viability, complete revascularization, number of bypass grafts, and an elective operation. CONCLUSIONS: The coronary vascular system can be described by means of a simple scoring system. A good or at least moderate coronary artery perfusing an area of dysfunctional yet viable myocardium is the main predictor of a successful perioperative course and an event-free survival. Patients with a poor coronary vasculature regardless of myocardial viability should not be considered suitable for revascularization. PMID- 12735555 TI - Radial artery use is safe in patients with moderate to severe left ventricular dysfunction. AB - BACKGROUND: Using radial artery grafts in patients with moderate to severe left ventricular dysfunction (LVD; ejection fraction < 35%) has been discouraged for the fear that postoperative vasopressor support may cause graft spasm and lead to ischemic complications. We, therefore, examined the safety of radial grafts in aortocoronary bypass (ACB) patients with LVD. METHODS: Data were collected from 5,455 patients who underwent isolated ACB between January 1995 and September 2001. One thousand eight hundred three patients received a radial artery graft (RadACB), and 3,652 patients did not (NoRadACB). Three hundred seven RadACB, and 819 NoRadACB operations were performed in LVD patients. A matched (age, sex, urgency of operation, diabetes, and renal insufficiency) cohort analysis was performed in LVD patients. Univariate and logistic regression analyses were performed in the entire population and the unmatched RadACB and NoRadACB patient subgroups to examine the effect of radial artery use on postoperative death or myocardial infarction rate. RESULTS: The matched cohort analysis revealed a similar rate of death or myocardial infarction (RadACB, 11 of 242 patients; NoRadACB, 16 of 242 patients; p = 0.32). Left ventricular dysfunction was associated with a higher rate of death or myocardial infarction in both unmatched groups (RadACB, odds ratio, 2.36; 95% confidence interval, 1.38 to 4.58; p = 0.004; NoRadACB, odds ratio, 1.62; 95% confidence interval, 1.18 to 2.24; p < 0.001) and in the entire population (odds ratio, 1.77; 95% confidence interval, 1.32 to 2.35; p = 0.003). An interaction term for patients with LVD and a radial artery graft, which was forced into the logistic regression model for the entire population, was not predictive of death or myocardial infarction (odds ratio, 1.52; 95% confidence interval, 0.75 to 3.10; p = 0.25). CONCLUSIONS: Left ventricular dysfunction carries similar risk for postoperative death or myocardial infarction in RadACB and NoRadACB patients. The presence of LVD in isolation is not a contraindication to the use of radial grafting. PMID- 12735556 TI - Supraclavicular control of patent internal thoracic artery graft flow during aortic valve replacement. AB - BACKGROUND: The mortality and morbidity of aortic valve replacement (AVR) after prior coronary artery bypass surgery (CABG) with patent left internal thoracic artery (LITA) is significant. The risk of LITA injury and inadequate myocardial preservation during the cross-clamp period may cause myocardial pump failure. METHODS: A total of 43 patients with a patent LITA graft underwent AVR. The patients were divided into the two groups. Group 1 included 19 patients who underwent AVR with deep hypothermia (20 degrees C) without LITA clamping. Group 2 included 24 patients in whom LITA flow was controlled through supraclavicular occlusion and AVR performed with moderate hypothermia (28 degrees C). RESULTS: Average cardiopulmonary bypass time (CPB) time was 118.79 +/- 20.36 minutes in group 1 and 102.67 +/- 9.66 minutes in group 2 (p = 0.006). Average cross-clamp time was 53.79 +/- 7.26 minutes in group 1 and 49.63 +/- 6.7 minutes in group 2 (p = 0.022). Inotropic support was required in 12 patients in group 1 and 4 patients in group 2 (p = 0.002). Average intensive care unit stay was 4.68 +/- 2.24 days in group 1 and 2.29 +/- 0.46 days in group 2 (p < 0.001). Average hospital stay was 11.84 +/- 2.91 days in group 1 and 8.04 +/- 2.38 days in group 2 (p < 0.001). Mortality due to myocardial failure developed in 4 patients in group 1 but in none of the patients in group 2 (p = 0.02). CONCLUSIONS: Proximal control of LITA flow by extrathoracic supraclavicular occlusion reduces the incidence of myocardial failure due to nonhomogenous cardioplegia delivery to the anterior wall of the heart, resulting in improved myocardial protection and the elimination of the need for deep hypothermia. PMID- 12735557 TI - Does skeletonization compromise the integrity of internal thoracic artery grafts? AB - BACKGROUND: There are few reports that demonstrate the chronologic changes in the functional integrity of the internal thoracic artery (ITA) wall after skeletonization. We investigated the impact of skeletonization on ITA wall integrity by immunohistochemical analyses in acute and chronic phases. METHODS: Nine mongrel dogs underwent bilateral ITA dissection with one skeletonized vessel and the other pedicled. The following studies were performed 1 week (acute phase, n = 3) and 12 weeks (chronic phase, n = 6) after ITA harvesting. All specimens of the ITAs were stained by antibodies against von Willebrand Factor (VWF), endothelial nitric oxide synthase (eNOS), inducible nitric oxide synthase (iNOS), and proliferating cell nuclear antigen (PCNA). After observation with confocal laser scanning microscopy, quantitative analyses of the staining signal for VWF and eNOS expressed on endothelial cells were performed. RESULTS: There were significantly more microvessels positive for VWF in the adventitia of skeletonized ITAs than in the adventitia of pedicled ITAs but the expression of PCNA in both groups was minimal, as in normal vessels. iNOS was not detected in any specimen. The intensity of VWF and eNOS expressed by endothelial cells had no significant differences between groups at either phase. CONCLUSIONS: The functional integrity of skeletonized ITA was similar to that of pedicled ITA in both acute and chronic phases. Although skeletonization induced neovascularization in the adventitia it did not induce proliferation of smooth muscle cells in the media, which is supposed to be a feature of vascular remodeling. PMID- 12735558 TI - Pitfalls and key lessons with the symmetry proximal anastomotic device in coronary artery bypass surgery. AB - BACKGROUND: Increasing practice of less-invasive coronary artery bypass surgery has generated a revival of automated methods to facilitate the creation of vascular connections during coronary artery bypass grafting procedures. METHODS: We have reviewed our clinical experience with 107 patients who received at least one proximal Symmetry aortic connector from St. Jude Medical Anastomotic Technology Group (St. Paul, MN) to connect a saphenous vein graft to the ascending aorta, in whom the minimal follow-up is 6 months. Seventy-five patients were part of an observational study, and 32 patients were enrolled in a prospective randomized study to compare the anastomotic device with the conventional suture technique. Key lessons and pitfalls observed during loading and deployment as well angiographic findings are presented. RESULTS: Hospital mortality was 0.9% (1/107); a 76-year-old woman suffered from cerebral complications and died after combined coronary artery bypass grafting and aortic valve replacement. Two connectors had to be removed because of leakage and one because of incomplete deployment. The residual 104 grafts connected to the ascending aorta were patent at the end of the operation (mean flow, 72 +/- 29 mL/min). Fourteen patients from the prospective study have received 6 months of angiographic assessment so far. Forty-two grafts have been evaluated: all IMA grafts (n = 14) are patent. Four radial artery grafts have been studied: three are patent and one is occluded. Twenty-four vein grafts have been assessed: 11 were hand-sewn and 13 were connected to the aorta with the proximal connector system. Patency rate is not different between the two techniques, but there is an incidence of 38% (5/13) stenosis in the proximal vein graft segment in those patients with a proximal connector. No proximal hand-sewn anastomosis shows any significant stenosis. CONCLUSIONS: The use of proximal devices allows ultrafast connection of a bypass graft to the aorta. The handling of this device is simple, manipulations on the ascending aorta are minimized, and hemostasis is perfect. However, because stenoses have been observed in the vein graft close to the connector site, mid- and long-term patency rates have to be assessed in large prospective trials. PMID- 12735559 TI - Nitric oxide donating aspirins: novel drugs for the treatment of saphenous vein graft failure. AB - BACKGROUND: A new class of nitric oxide donating aspirin (NO-ASA) drugs may increase the therapeutic impact of aspirin in saphenous vein coronary artery bypass grafting (CABG) not only through the inhibition of thrombosis but also through a reduction of vasospasm and inhibition of vascular smooth muscle cell (VSMC) proliferation (effects that are inhibited by NO but not ASA). In order to test this proposal the effect of three NO-ASA drugs (NCX4040, NCX4050, and NCX4060) on in vitro relaxation and cyclic guanosine monophosphate (cGMP) formation in the human isolated saphenous vein and the proliferation of human VSMCs was investigated. METHODS: Saphenous vein segments were obtained from 30 patients undergoing CABG (median age, 59 years; range, 49 to 68). The effect of the NO-ASA adducts, ASA alone, and sodium nitroprusside (NO donor) were investigated on (1) relaxation of phenylephrine-stimulated contraction using an organ bath, (2) cyclic guanosine monophosphate (cGMP) formation using an enzyme linked immunosorbent assay, and (3) the proliferation of VSMCs derived from saphenous vein using bromo-deoxyuridine (BRDU) incorporation. RESULTS: All three NO-ASA adducts (at concentrations that inhibited responses by 50% [IC50s] between 1 micromol/L and 100 micromol/L) and nitroprusside (at IC50s between 0.5 and 10 micromol/L) elicited relaxation of isolated human saphenous vein, promoted cGMP formation, and inhibited VSMC proliferation whereas ASA alone (up to 100 micromol/L) had no effect on any variable. CONCLUSIONS: These data indicate that the NO-ASA adducts by virtue of their capacity to release NO and stimulate guanylyl cyclase may be useful not only in the prevention of thrombosis following CABG but also the reduction of saphenous vein graft spasm and neointima formation. PMID- 12735560 TI - Endogenous myocardial angiogenesis and revascularization using a gastric submucosal patch. AB - BACKGROUND: The gastrointestinal submucosa physiologically produces angiogenic proteins. We examined whether these properties could lead to endogenous myocardial angiogenesis in a swine model of chronic ischemia. METHODS: Fifteen Yorkshire swine underwent ameroid constrictor placement around the circumflex artery and either lateral epicardial abrasion, creation of a gastroepiploic artery (GEA) based gastric patch, mucosal avulsion, transdiaphragmatic transfer, and apposition of the patch against the circumflex myocardial territory (number = 8; test animals), or lateral epicardial abrasion alone (number = 7; controls). Seven weeks later, lateral myocardial perfusion, endothelial cell density, and expression of VEGFR-1 and VE-cadherin were determined using isotope-labeled microsphere assays, immunohistochemistry, and immunoblotting, respectively. RESULTS: Microsphere assays showed equivalent lateral/anterior myocardial perfusion indices at rest (1.10 +/- 0.49 vs 0.95 +/- 0.23, test vs control animals; p = 0.54), but higher perfusion in test animals versus controls during pacing (1.05 +/- 0.29 vs 0.69 +/- 0.09, test vs controls; p = 0.02). Increased myocardial endothelial cell density (42.6 +/- 8.5 vs 26.1 +/- 11.6 cells per 3850 microm2, test vs controls; p = 0.02) and expression of VE-cadherin (3.10 +/- 0.60 fold change, test vs controls; p = 0.001) were also observed in the lateral territory of test animals versus controls. Reconstitution of the proximally occluded circumflex artery from patch collaterals was demonstrated on gastroepiploic arteriography in a subset of test animals. CONCLUSIONS: This model results in an angiogenic process of significantly greater magnitude than that resulting from chronic myocardial ischemia alone, without the need for exogenous angiogenic agents. PMID- 12735562 TI - In vivo repopulation of xenogeneic and allogeneic acellular valve matrix conduits in the pulmonary circulation. AB - BACKGROUND: Approaches to in vivo repopulation of acellularized valve matrix constructs have been described recently. However, early calcification of acellularized matrices repopulated in vivo remains a major obstacle. We hypothesised that the matrix composition has a significant influence on the onset of early calcification. Therefore, we evaluated the calcification of acellularized allogenic ovine (AVMC) and xenogenic porcine (XVMC) valve matrix conduits in the pulmonary circulation in a sheep model. METHODS: Porcine (n = 3) and sheep (n = 3) pulmonary valve conduits were acellularized by trypsin/EDTA digestion and then implanted into healthy sheep in pulmonary valve position using extracorporeal bypass support. Transthoracic echocardiography (TTE) was performed at 12 and 24 weeks after the implantation. The animals were sacrificed at week 24 or earlier when severe calcification of the valve conduit became evident by TTE. The valves were examined histologically and biochemically. RESULTS: All AVMC revealed severe calcification after 12 weeks with focal endothelial cell clustering and no interstitial valve tissue reconstitution. In contrast, after 24 weeks XVMC indicated mild calcification on histologic examination (von Kossa staining) with histologic reconstitution of valve tissue and confluent endothelial surface coverage. Furthermore, immunohistologic analysis revealed reconstitution of surface endothelial cell monolayer (von Willebrand factor), and interstitial myofibroblasts (Vimentin/Desmin). CONCLUSIONS: Porcine acellularized XVMC are resistant to early calcification during in vivo reseeding. Furthermore, XVMC are repopulated in vivo with valve-specific cell types within 24 weeks resembling native valve tissue. PMID- 12735561 TI - Development of abnormal tissue architecture in transplanted neonatal rat myocytes. AB - BACKGROUND: Most myocardial cell transplant studies focus on demonstration of improved function; however, such improvement depends on the development of appropriate tissue structure. Thus, our aim was to assess the architectural changes that occurred after cell transplant into normal and infarcted myocardium. METHODS: Male neonatal cells (1 to 2 days old) were injected into the left ventricular free wall of adult female rats. The tissue was examined 0 to 1 days and 1 to 2, 4 to 6, and 12 weeks later in noninfarcted hearts and 6 months after transplant into infarcts. In histologic sections, we assessed the cells' retardation of polarized light (to measure development of contractile elements), two-dimensional cell orientation, cell nuclear morphology, and collagen content. RESULTS: The transplant cells' retardation of polarized light gradually increased to 81% of that of host cells after 6 months (p < 0.001). The transplant cells were disorganized and although their nuclei increased in size, they always had a rounded appearance. Collagen content in the transplant was 210% to 430% higher than in host tissue (p < 0.01). In addition, scar collagen always separated transplant and host cells. CONCLUSIONS: One architectural feature, the rounded nuclei, provided a distinctive marker to identify transplanted cells. Nevertheless, the transplants' inhibited muscle development together with disorganization, separation from the host muscle, and a substantial increase in collagen resulted in a structure unlikely to play an active role in systolic function. PMID- 12735563 TI - Right ventricular cardiomyoplasty: 10-year follow-up. AB - BACKGROUND: Chronically depressed right ventricular (RV) function presents an unsolved therapeutic challenge in cardiac surgery. Despite recent advances in medical and surgical therapies, prognosis remains poor and patient's quality of life and mortality are frequently unacceptable. The aim of this study is to present the first clinical report and long-term results of RV dynamic cardiomyoplasty applied in patients with RV failure caused by isolated RV cardiomyopathies. METHODS: Seven consecutive patients (5 males, 2 females; mean age, 40 +/- 9 years; range, 15 to 63 years) from a series of 113 cardiomyoplasty procedures performed at Broussais and Pompidou Hospitals were evaluated. The mean duration of follow-up was 10 +/- 3.5 years. All patients had predominant RV dysfunction, associated with tricuspid regurgitation in 6 patients. The cause of RV failure was arrhythmogenic cardiomyopathy (4 patients), ischemic (2 patients), and Uhl's disease (1 patient), and endomyocardial fibrosis (1 patient). Six patients were in preoperative New York Heart Association functional class III and 1 was in intermittent class III/IV. The mean preoperative ejection fraction (measured by isotopic technique) was 18% +/- 5.7% for the right ventricle and 40% +/- 13% for the left ventricle. Right ventricular dynamic cardiomyoplasty consists of wrapping the RV free walls with the left latissimus dorsi muscle flap. The distal part of the latissimus dorsi muscle is fixed to the diaphragm and then electrostimulated. Six patients required associated tricuspid valve surgery. RESULTS: There were no perioperative deaths. The mean duration of follow up was 10 +/- 3.5 years. Six patients are alive with a remarkable quality of life, 4 are in New York Heart Association functional class I and 2 are in class II. One patient who was in New York Heart Association functional class II died in postoperative year 7 caused by stroke. At last follow-up, mean RV ejection fraction was 33% +/- 11.8% and left ventricular ejection fraction was 52% +/- 12.6%. CONCLUSIONS: The results of this long-term study demonstrate hemodynamic and functional improvements after RV cardiomyoplasty without perioperative mortality, no long-term malignant arrhythmias, and RV dysfunction related deaths. We believe that RV cardiomyoplasty, associated with tricuspid valve surgery when required, could be an effective treatment for severe RV failure. PMID- 12735565 TI - Three or more median sternotomies for patients with valve disease: role of computed tomography. AB - BACKGROUND: We evaluated the effects of computed tomographic (CT) scan-guided third or fourth median sternotomies for valve operations on the incidence of resternotomy-related complications and early mortality. METHODS: Ninety patients undergoing valve replacement with third or fourth time sternotomy were divided into two groups. One group (CT group) consisted of 64 patients who had undergone routine CT scans preoperatively after 1991 to assess the possibility of sternotomy-related bleeding, and the other group (no CT group) comprised the remaining 26 patients who did not receive CT scans. RESULTS: Hospital death occurred in 4 patients (6%) in the CT group and in 6 patients (23%) in the no CT group (p = 0.0309). Multivariate analysis indicated NYHA class 4 (odds ratio [OR] = 6.99) and year of operation (OR = 1.05) to be predictors of hospital death. Preoperative CT scans revealed that 8 patients were considered to be high risk for resternotomy, they underwent femorofemoral bypass before sternal division was performed. Hemorrhage occurred upon sternal reentry in 2 of these 8 patients. The incidences of sternotomy-related injury were 19% (5/26) in the no CT group and only 3% (2/64) in the CT group (p = 0.0198). Multivariate analyses demonstrated a fourth sternotomy (OR = 4.31) to be a predictor of resternotomy-related injury. CONCLUSIONS: CT scans provide preoperative information on retrosternal adhesions. When a distended heart or expanded aorta has adhered to the sternum, femorofemoral cannulation should be performed before sternotomy. Third and fourth sternotomies, though demanding procedures, can be performed safely using the described strategy. PMID- 12735564 TI - Preliminary experience with the LionHeart left ventricular assist device in patients with end-stage heart failure. AB - BACKGROUND: The Arrow LionHeart LVD 2000 left ventricular assist device is the first fully implantable system designed for destination therapy. We report on 2 years of experience with this device, which we implanted for the first time in October 1999. METHODS: Since October 1999, 6 male patients between 55 and 69 years of age (mean 65 +/- 6 years) have received the device at our center; all were in New York Heart Association functional class IV and ineligible for heart transplantation. RESULTS: All surgical procedures were uneventful, with a timely extubation in 5 of 6 patients. Duration of support was 17 to 670 (mean 245 +/- 138) days, with a cumulative experience of 4.5 years. Three patients recovered to be discharged from hospital under support and are long-term survivors. Three patients died 17, 31, and 112 days after implantation from multiple organ failure without being discharged to their homes. The survival rate is 50% after 18 months. There were no major system-related problems or any device-related infections, which are otherwise commonly found among vertricular assist device patients. CONCLUSIONS: Our preliminary experience demonstrates the reliability and efficacy of the different parts of the system. Nevertheless, further sophistication is needed to reduce the size of its components, which so far still constitutes a limiting factor. PMID- 12735566 TI - Prognosis after aortic root replacement with cryopreserved allografts in adults. AB - BACKGROUND: Aortic root replacement with cryopreserved allografts is associated with excellent hemodynamics, little endocarditis, low thromboembolic event rates, and no need for anticoagulation. There is, however, concern regarding the long term durability of this valve substitute, especially in younger patients. Meta analysis and microsimulation were used to calculate age-specific long-term prognosis after allograft aortic root replacement based on current evidence. METHODS: Our center's experience with cryopreserved allograft aortic root replacement in 165 adult patients was combined in a meta-analysis with reported and individual results from four other hospitals. Using this information, the microsimulation model predicted age- and gender-specific total and reoperation free and event-free life expectancy. RESULTS: The pooled results comprised 629 patients with a total follow-up of 1860 patient-years (range 0 to 12.8 years). Annual risks were 0.6% for thromboembolism, 0.05% for bleeding, 0.5% for endocarditis, and 0.5% for nonstructural valve failure. Structural allograft failure requiring reoperation occurred in 15 patients, and a patient age-specific Weibull function was constructed accordingly. Calculated total life expectancy varied from 27 years in a 25-year-old to 12 years in a 65-year-old male; corresponding actual lifetime risk of reoperation was 89% and 35%, respectively. CONCLUSIONS: Cryopreserved aortic allografts have an age-related limited durability. This results in a considerable lifetime risk of reoperation, especially in young patients. The combination of meta-analysis and microsimulation provides an appropriate tool for estimating individualized long term outcome after aortic valve replacement and can be useful both for patient counseling and prognostic research purposes. PMID- 12735567 TI - Left atrial maze procedure: a useful addition to other corrective operations. AB - BACKGROUND: The left atrial maze procedure is performed to treat atrial fibrillation (AF), mainly in patients with mitral valve disease. In this study, we assessed the midterm results of this procedure and clinically analyzed predicting factors for postoperative persistent AF. METHODS: From June 1997 to May 2001, the left atrial maze procedure was performed on 31 patients (29 with mitral valve disease and 2 lone AF). For purposes of analysis, patients were divided postoperatively into those with persistent atrial fibrillation (AF) and those with sinus rhythm (SR), except 2 patients who required pacemaker implantation for sinus node dysfunction. Over a follow-up period of more than 12 months, patients were compared based on their preoperative and intraoperative variables. RESULTS: At discharge, the success rate was 89.7%. The midterm rates (total of 94.9 patient-years of follow-up) of sinus rhythm and freedom from AF were 72.4% and 79.3%, respectively. There were significant differences in duration of AF, voltage of f-wave at first precordial lead of electrocardiogram, and cardiothoracic ratio between the SR and AF groups. CONCLUSIONS: Our midterm results suggest that the left atrial maze procedure is an effective alternative adjunct procedure for elective open heart surgery to treat AF, depending upon the patient's clinical condition. PMID- 12735568 TI - Intra-atrial temperatures in radiofrequency endocardial ablation: histologic evaluation of lesions. AB - BACKGROUND: Because of the limited information on the effects of ablation in human tissues, we studied intra-atrial temperatures during endocardial radiofrequency applications. We correlated the intra-tissue temperatures with the tissue thickness and with the histologic appearance of the lesions. METHODS: Radiofrequency currents were delivered to human atrial tissue, simulating conditions in endocardial ablation during surgery at set temperature of 70 degrees and 80 degrees C, and intra-tissue temperatures were measured with thermocouples. Radiofrequency applications at 70 degrees C were performed in patients undergoing mitral valve surgery and biopsy specimens were obtained. Samples from in vitro studies and from patients were assessed histologically. RESULTS: The subepicardial temperatures were usually over 60 degrees C in applications in vitro at 70 degrees C and over 70 degrees C in applications at 80 degrees C. Values were higher when the interior of the tissue was warmer than its surface as a result of consecutive radiofrequency applications over the same area. Histologic examination of 12 in vitro samples showed that 10 had transmural lesions. Five of 10 samples from patients with mitral valve surgery had lesions confined to the endocardium, 3 had damaged variable portions of the myocardium, and 2 had transmural lesions. CONCLUSIONS: Although it is possible to obtain transmural lesions in vitro and in vivo with endocardial applications at 70 degrees C, it is significantly more difficult to achieve transmural lesions in patients with mitral valve disease than in normal atrial tissue in vitro. Consecutive applications can raise the intra-tissue temperatures to values significantly higher than those used for application. Our findings suggest that the composition of the endocardium and of the myocardium is a major determinant in lesion formation. PMID- 12735569 TI - The conduction system in hearts with pulmonary atresia and intact ventricular septum. AB - BACKGROUND: Although a regular course of the conduction system is anticipated in hearts with pulmonary atresia and intact ventricular septum (PAIVS), it has never been demonstrated anatomically. In view of one of the surgical options in repair being debulking of the right ventricular wall, it is important to establish the location of the major conduction pathways. METHODS: Four hearts belonging to fetuses aged 18, 20, 22, and 29 weeks were examined. Entire hearts were serially sectioned and step sections were stained. The sinus node and atrioventricular conduction bundles were identified and their locations described. RESULTS: The conduction system in hearts with PAIVS is similar to that in normal hearts, although there was a minor variation in one of them. This was a heart affected by Ebstein malformation of the tricuspid valve. In this case, the right bundle branch terminated abruptly soon after its origin. CONCLUSIONS: Confirmation of the regular course of the cardiac conduction system is important to the development of strategies for surgical repair of PAIVS. PMID- 12735570 TI - Blood loss in infants and children for open heart operations: albumin 5% versus fresh-frozen plasma in the prime. AB - BACKGROUND: Infants and children undergoing cardiopulmonary bypass become substantially hemodiluted secondary to the volume used to prime the oxygenator. Fresh-frozen plasma has been included in the prime to lessen dilution of clotting factors and correspondingly minimize blood loss and transfusions. METHODS: We prospectively randomized 56 patients weighing 10 kg or less who required cardiopulmonary bypass to receive either one unit of fresh-frozen plasma or 200 mL of albumin 5% in the prime. After protamine administration, samples for prothrombin time, fibrinogen, platelet count, and thromboelastogram were obtained. Mediastinal chest tube drainage and transfusion requirements were documented. RESULTS: There were no significant differences between groups regarding demographic or surgical characteristics. Blood loss during the first 24 hours was similar in both groups, but total transfusions were significantly greater in those who received fresh-frozen plasma instead of albumin 5% in the prime (8.0 +/- 4.2 versus 6.1 +/- 4.5 U, respectively; p = 0.035). Post hoc analyses suggest that for cyanotic patients and patients undergoing complex operations, fresh-frozen plasma in the prime results in less blood loss than albumin 5%. CONCLUSIONS: Substitution of albumin 5% for fresh-frozen plasma in the prime of acyanotic patients weighing 10 kg or less who undergo noncomplex operations requiring cardiopulmonary bypass significantly reduces perioperative transfusions without increasing blood loss. Further investigation is needed to determine whether increased blood loss is associated with increased transfusions when albumin 5% is substituted for fresh-frozen plasma in the prime of infants and children who are cyanotic or undergoing complex operations. PMID- 12735571 TI - Management of traumatic rupture of the thoracic aorta in pediatric patients. AB - BACKGROUND: Traumatic rupture of the thoracic aorta (TRA) in the pediatric population is uncommon. Management of TRA in general has evolved to include selective nonoperative and endovascular stent graft approaches, although operative repair remains the standard. METHODS: We conducted a retrospective chart review of patients younger than 16 years of age admitted to a single institution between March 1985 and February 2002. RESULTS: Of 160 patients admitted with TRA, 11 were younger than 16 (11.9 +/- 3.5) years of age. Concomitant injuries included closed head injury (5 patients) and acute lung injury (6 patients). All were started on beta-blockers when the diagnosis was suspected. Laparotomy was required in 3 patients and orthopedic procedures in 5 patients. Six underwent operative repair (two primary repairs), with no mortality. Cross-clamp time was 30.4 +/- 2.6 minutes. One patient (operated on without bypass) was partially paralyzed. Two patients were managed nonoperatively, 1 with an intimal arch injury, who on subsequent follow-up has demonstrated healing, and 1 who died of head injury. Three patients were managed by endovascular stent grafts, 2 who died of closed head injury and 1 who at 1 year follow-up has fully recovered. The endovascular stent grafts were placed through the femoral artery in 2 patients and through an iliac conduit in 1 patient. No patient died of rupture. CONCLUSIONS: The approach to pediatric TRA should be identical to the adult, with early institution of beta-blockers. Depending on the clinical setting, a spectrum of options should be considered, including operation, non-operation, and endovascular stent graft, although the choice of the latter must be tempered by the lack of long-term follow-up data. PMID- 12735572 TI - Valve sparing aortic root replacement for dilatation of the pulmonary autograft and aortic regurgitation after the Ross procedure. AB - BACKGROUND: Aortic insufficiency secondary to progressive dilatation of the pulmonary autograft is being recognized with increasing frequency after the Ross procedure. We reviewed our experience with valve-sparing aortic root replacement concomitant with aortic annuloplasty to assess the effectiveness of this approach. METHODS: Four patients, aged 8 to 27 years, presented with moderate to severe aortic insufficiency associated with progressive root dilatation from 1 to 8 years after a Ross procedure. All patients had 0 to 1+ aortic insufficiency early after the Ross procedure, with a mean maximal sinus diameter of 37 mm (range 30 to 45 mm). At reoperation the maximum diameter of the root ranged from 45 to 55 mm (mean 50 +/- 4 mm). A valve-sparing aortic root replacement with annular reduction was performed. The annulus was decreased from a mean of 27 mm to 23 mm. For the root replacement, 1 patient underwent a standard root remodeling procedure; in the others, a separate piece of scalloped Dacron (C.R. Bard, Haverhill, PA) graft material was used for each sinus to facilitate optimal exposure. RESULTS: All 4 patients are in New York Heart Association functional class I at a mean follow-up of 6 months. The most recent echocardiography demonstrated 0 to 1+ aortic insufficiency with good left ventricular function. Histology of the excised pulmonary autograft walls demonstrated severe elastin fragmentation. CONCLUSIONS: Aortic root remodeling with annular reduction is an effective treatment for aortic root dilatation and aortic insufficiency after the Ross operation. This procedure allows correction of aortic insufficiency and avoids the need for a prosthetic valve and anticoagulation. PMID- 12735574 TI - Secundum ASD closure using a right lateral minithoracotomy: five-year experience in 122 patients. AB - BACKGROUND: Surgical closure of secundum atrial septal defect (ASD) is a standard procedure associated with very low mortality and morbidity. We evaluated outcomes in the era of catheter-based interventional closure and minimally invasive techniques. METHODS: From May 1996, February 2002, 177 patients with a body weight of more than 30 kg underwent surgical ASD closure. A right lateral minithoracotomy (LMT) was used in 122 patients and a conventional approach, in 55. Diagnoses included secundum ASD in 106 patients in the LMT group and 40 in the conventional group, sinus venosus ASD in 13 patients in each group, and status post interventional closure in 3 and 2 patients, respectively. Mean age was 37 +/- 17 years in the LMT group and 43 +/- 20 years, in the conventional group and mean body weight was 66 +/- 17 kg and 70 +/- 16 kg, respectively. In the LMT group, femoral cannulation was performed for cardiopulmonary bypass. RESULTS: Direct ASD closure was carried out in 67.2% of patients in the LMT group and 58.2% of those in the conventional group. The remaining patients had pericardial patch closure. There was one death: A patient in the conventional group who required explantation of an Amplatzer device because of infection died postoperatively. Average stay in the intensive care unit was 1.2 +/- 0.5 days. Two patients required reoperation for residual ASD after direct closure; 1 sustained a temporary neurological deficit that resolved completely. On postoperative echocardiography, a minimal residual shunt was seen in only 3 patients. All patients were in good clinical condition with improved functional status at discharge from the hospital. CONCLUSIONS: Secundum ASD closure by LMT has become as standard and safe an operation as the conventional technique and achieves good perioperative results and satisfactory long-term outcomes. Thus LMT is an attractive option for patients who are not suitable for closure using catheter-based devices. PMID- 12735573 TI - Vascular endothelial growth factor in children with congenital heart disease. AB - BACKGROUND: Children with cyanotic congenital heart disease may experience the development of abnormal vessels that become a source of significant morbidity. Abnormal vessel proliferation in these children may take several forms, including systemic-to-pulmonary collateral arteries, systemic-to-pulmonary venous collaterals, systemic venous collateral channels after bidirectional cavopulmonary anastomosis, and pulmonary arteriovenous malformations. However, no entity responsible for these abnormalities has been identified yet. This study determined whether children with cyanotic congenital heart disease have elevated serum levels of vascular endothelial growth factor (VEGF) and whether elevated VEGF correlated with these abnormal vessels. METHODS: Mean systemic room air oxygen saturation (SpO2), blood cell counts (RBC), and serum VEGF levels were measured preoperatively. Samples were obtained from 61 children with acyanotic heart disease (group N) and 102 children with cyanotic heart disease (group C) before cardiac surgery. Postoperative catheterization was performed 1-month after the operation to evaluate the abnormal vessels in group C. RESULTS: The VEGF level was significantly elevated in group C (355.0 +/- 287.1 pg/mL) compared with group N (203.0 +/- 221.6 pg/mL; p < 0.001). VEGF levels in patients with a single ventricle associated with asplenia syndrome (n = 7) in group C were significantly elevated (711.9 +/- 443.5 pg/mL) compared with other patients. There was no significant correlation between VEGF level and SpO2 or RBC. Abnormal vessels were diagnosed in 19.6% (20/102) patients in group C. There was no difference in VEGF levels between the patients with abnormal vessels (336.8 +/- 182.5 pg/mL) and the patients without abnormal vessels (359.1 +/- 306.8 pg/mL). CONCLUSIONS: Children with cyanotic heart disease have elevated systemic levels of VEGF, especially in those patients with a single ventricle associated with asplenia syndrome. There was no significant relationship in VEGF levels between the patients with abnormal vessels and without these vessels. PMID- 12735575 TI - Safety and efficacy of minimally invasive atrial septal defect closure. AB - BACKGROUND: Atrial septal defects (ASDs) have been surgically closed with low mortality utilizing the conventional sternotomy approach (CSA). The technical ease of ASD closure has triggered interest in minimally invasive closure (MIC) to obviate the morbidity associated with sternotomy. Our study assesses the safety and efficacy of minimally invasive ASD closure. METHODS: Preoperative, intraoperative, and postoperative data were collected on 68 patients (39 CSA, 29 MIC) who underwent ASD closure from January 1997 to August 2002. Using univariate analysis of 17 preoperative risk factors there was no statistically significant difference between the two groups. RESULTS: MIC resulted in equivalent success rates in ASD closures, with similar morbidity, no mortality, and a significant difference in postoperative length of stay (3.93 +/- 1.6 days versus 5.36 +/- 2.51 days, p = 0.006). CONCLUSIONS: In experienced hands, MIC is an excellent alternative to CSA in ASD closure. PMID- 12735576 TI - Anatomic repair of anomalous left coronary artery from the pulmonary artery by aortic reimplantation: early survival, patterns of ventricular recovery and late outcome. AB - BACKGROUND: To determine the early and late outcomes of patients presenting with anomalous left coronary artery from the pulmonary artery who had repair by aortic reimplantation. METHODS: From January 1952 to July 2000, 67 patients presented with anomalous coronary artery from the pulmonary artery. Forty-seven patients who had repairs performed by aortic reimplantation are the subject of this study. The median age at repair was 7.7 months. Before repair, 10 infants (21%) presented in extremis requiring ventilatory and inotropic support, and 38 infants (80%) presented in heart failure. Autologous pericardial hood coronary arterioplasty was used in 4 patients, and concomitant mitral valve repair was used in 1 patient. RESULTS: Hospital survival was 92%. Five children required postoperative extracorporeal membrane oxygenation for a median of 4 days (range, 2 to 8 days). Patients who had extracorporeal membrane oxygenation were significantly more likely to have presented in critical condition (40% vs 3% if no extracorporeal membrane oxygenation; p = 0.006) or with ventricular arrhythmias (67% vs 7%; p = 0.027), to have presented with significantly lower preoperative repair median ejection fraction (10%, n = 5 vs 40%, n = 38; p = 0.01) or to have presented with more severe left ventricular dilatation (p = 0.03). Within a 15-year or less follow-up (mean, 4.7 years) there were no late deaths. Kaplan-Meier survival was 91% at 5 years, and freedom from reoperation was 93% at 10 years. At late follow-up, echocardiography demonstrated significant improvements in mean ejection fraction (64% +/- 9% vs 33% +/- 21% preoperatively, p < 0.0001); moderate mitral regurgitation (9% vs 38% preoperatively, p < 0.02); and wall motion abnormalities (15% vs 81% preoperatively, p < 0.002). The ratio of measured left ventricular end-diastolic dimension to the 95th percentile of normal declined from 1.4 +/- 0.3 to 1.0 +/- 0.1 (p < 0.0006). Children who had extracorporeal membrane oxygenation had normal ejection fractions and ventricular dimensions at follow-up (n = 3). Repeated measures of mixed linear regression analysis demonstrated that normalization of ejection fraction and left ventricular function occurred within 1 year of repair. Improvements in mitral regurgitation lagged behind normalization of ejection fraction and left ventricular dilatation. CONCLUSIONS: Anatomic repair of anomalous left coronary artery from the pulmonary artery by aortic reimplantation yields excellent early survival and late functional outcomes even in critically ill infants. PMID- 12735577 TI - Contrast echocardiography: potential for the in-vivo study of pediatric myocardial preservation. AB - BACKGROUND: Myocardial contrast echocardiography (MCE) has been used successfully during adult cardiac surgery to image myocardial perfusion. Recently it has been suggested this technique is capable of detecting microvascular injury and inflammation because sonicated albumin microbubbles adhere to activated neutrophils and, in the presence of denuded or inflamed endothelium, they persist within the microvasculature rather than passing unimpeded, which results in profound slowing of their transit rates. The technique has not previously been used during congenital heart surgery; however significant potential is suggested in this setting in which myocardial inflammation may contribute to postoperative myocardial dysfunction, a leading cause of morbidity and mortality. We have performed a preliminary study to assess the safety and feasibility of MCE in the pediatric intraoperative environment and to examine myocardial transit rates. METHODS: Sonicated albumin microbubbles were injected with cardioplegia during bypass in 16 children (aged 3 weeks to 8.5 years). Images were collected using transesophageal echocardiography. Complications, post-bypass electrocardiographic, echocardiographic, and outcome data were recorded. Myocardial transit rates were calculated using videointensity analysis, assessed for reproducibility and correlated with demographic and intraoperative variables and postoperative outcome. RESULTS: The technique was performed safely, with good reproducibility. Myocardial persistence of microbubbles, which occurred in 6 patients, was associated with crystalloid cardioplegia, prolonged preischemic bypass (r = 0.72, p = 0.004), or ischemic time (r = 0.69, p = 0.002). CONCLUSIONS: Intraoperative MCE shows potential as an in vivo technique for the study of pediatric myocardial preservation. PMID- 12735578 TI - Preoperative high leukocyte count: a novel risk factor for stroke after cardiac surgery. AB - BACKGROUND: Stroke after cardiac surgery is a devastating complication. The relationship between white blood cell count (WBC) and perioperative cerebrovascular accident (CVA) has not been investigated. An effort was made to identify how preoperative WBC may relate to CVA development during or after cardiac surgery. METHODS: Prospective data were collected from 7,483 patients who underwent coronary artery bypass grafting or valvular surgery or both. WBC was determined preoperatively and postoperatively. Differentiation of WBC was examined only preoperatively. RESULTS: There were a total of 125 CVAs (10 transient ischemic attacks [TIAs], 115 strokes). WBC was significantly higher preoperatively and directly postoperatively in patients with stroke. Qualitative changes in preoperative WBC were also found in these patients (chi2; p < 0.001). The predictive power of the stepwise logistic regression model for CVA was greater when preoperative WBC was included. The risk for perioperative CVA increased starting at preoperative WBC of 9 x 10(9)/L (p = 0.044) and progressed in higher WBC ranges. WBC had a significant impact on CVA outcome (analysis of variance, p = 0.001). CONCLUSIONS: Our studies have established the correlation between high preoperative WBC and stroke during or after cardiac surgery. Furthermore, elevated preoperative WBC was related to the clinical outcome of CVA. Preoperative measures aimed at preventing or treating conditions such as infections that may cause elevated WBC may be beneficial in the prevention of stroke during or after cardiac surgery. PMID- 12735579 TI - Influence of prolonged cardiopulmonary bypass times on splanchnic perfusion and markers of splanchnic organ function. AB - BACKGROUND: Cardiopulmonary bypass (CPB) is known to have considerable negative impact on perfusion and organ function. The effects of the duration of CPB on markers of splanchnic organ function was studied. METHODS: Consecutive patients undergoing elective aorto-coronary bypass grafting with CPB times (CPBT) of either less than 70 minutes (n = 15) or more than 80 minutes (n = 15) were prospectively studied. Splanchnic perfusion was assessed by measuring arterial and gastric mucosal PCO2 and calculating PCO2gap. Hepatic function was evaluated by monoethylglycinexylidide (MEGX) test and by measuring alpha-glutathione S transferase (alpha-GST). Concentration of pancreatitis-associated protein was measured to assess pancreatic integrity. Measurements were performed after induction of anesthesia, at the end of surgery, 4 hours after arrival in the intensive care unit, and on postoperative day 1. RESULTS: The mean (+/- standard deviation) CPBT were 54 +/- 12 minutes and 99 +/- 16 minutes, respectively. PCO2gap increased significantly more in the group with CPBT of more than 80 minutes than in that with CPBT of less than 70 minutes, at +15 +/- 4 mm Hg versus +8 +/- 3 mm Hg, respectively, indicating reduction in splanchnic perfusion by longer CPBTs. Postoperative MEGX concentrations were significantly lower and postoperative alpha-GST concentrations were significantly higher in the group with CPBT of more than 80 minutes than in that with CPBT of less than 70 minutes. Plasma levels of pancreatitis-associated protein remained similar in both groups throughout the study period. CONCLUSIONS: In our patients with CPBT of more than 80 minutes, splanchnic perfusion and hepatocelluar integrity were moderately affected, whereas pancreatic function remained almost unchanged. Studies including a larger patient population are necessary to assess whether protective approaches would be helpful in patients undergoing complex cardiac surgery with very long CPBT. PMID- 12735580 TI - Mediastinitis in heart and lung transplantation: 15 years experience. AB - BACKGROUND: Mediastinitis after sternotomy carries a very high mortality, especially in patients receiving immunosuppressive treatment. METHODS: A retrospective analysis of the data for patients who had undergone cardiopulmonary transplantation between May 1985 and December 2000 was undertaken. A total of 776 patients had either a median sternotomy or a transverse sternotomy through a clam shell incision. Transplantations were as follows: 591 heart (3 simultaneous heart and renal, and 1 heart and liver), 126 bilateral sequential lung, 57 heart-lung, 1 en bloc double-lung, and 1 heart and single-lung. RESULTS: In all, 21 (2.7%) recipients had mediastinitis. Of these, 14 had heart, 3 heart-lung, and 4 bilateral lung transplantation. There were 18 median and 3 transverse sternotomies. There were 6 deaths (28.6%). Treatment consisted of antibiotics alone in 2 patients and subxiphisternal drainage in another 2 patients. The sternum was reopened in 17 (80.95%) patients, with debridement and primary closure alone in 5 of these 17 patients and additional irrigation in the other 12. Those who had resternotomy, debridement, and substernal irrigation had a better outcome when compared with the outcomes of other modes of treatment (1 death among 12 patients) (p = 0.06). Age, cardiopulmonary bypass time, body mass index, time to diagnosis, and treatment did not differ between those who survived and those who did not. CONCLUSIONS: Early aggressive debridement with substernal irrigation is the best mode of treatment for patients with posttransplantation mediastinitis. PMID- 12735581 TI - Long-term evaluation of the replacement of the trachea with an autologous aortic graft. AB - BACKGROUND: Tracheal reconstruction after extensive resection remains a challenge in thoracic surgery. The goal of this experimental study was to analyze the long term evolution of tracheal replacement using an autologous aortic graft. METHODS: In 21 sheep, a 5-cm segment of the cervical trachea was replaced by a segment of the descending thoracic aorta that was reconstructed to a prosthetic graft. Because of the airway collapse reported in a previous series, a permanent (n = 13) or temporary (n = 8) stent was systematically placed in the lumen of the graft. Clinical, bronchoscopic, and histologic examinations were performed up to 3 years after implantation. RESULTS: All animals survived the operation with no paraplegia. In the group with a permanent stent, three complications occurred: one stent displacement, one laryngeal edema, and one infection. Stent removal was tolerated after 6 months in the group with a temporary stent. Histologic examination showed a progressive transformation of the arterial segment into first extensive inflammatory tissue with a squamous epithelium, and after 6 to 36 months well-differentiated tracheal tissue including a continuous mucociliary epithelium and regular rings of newly formed cartilage. CONCLUSIONS: An autologous aortic graft used as a substitute for extensive tracheal replacement in sheep remained functional for periods up to 3 years. The progressive transformation of the graft into a structure resembling tracheal tissue seems to be a key factor in long-term patency. The mechanism of this regenerative process and the possibility of using arterial homografts, which would make clinical application easier, remain to be evaluated. PMID- 12735582 TI - Tracheobronchial obstruction in children: experience with endoscopic airway stenting. AB - BACKGROUND: We reviewed our experience to determine the role of endoscopic airway stents in children with tracheobronchial obstruction. METHODS: Seventeen children (10 boys and 7 girls) aged 2 months to 16 years underwent tracheobronchial stenting. Etiology of the tracheobronchial obstruction included external vascular compression (n = 9); tracheobronchial anastomotic strictures after heart lung/lung transplantation (n = 4); airway compression by malignant mediastinal mass (n = 2), and subglottic/high tracheal stenosis after prolonged intubation with a tracheostomy in situ (n = 2). Indications for airway stenting were failure to wean from ventilator after a mean of 82.5 days (range, 2 to 210) in 8 children; and dyspnea or stridor in the remaining 9 children. RESULTS: Ten children had a total of 24 uncovered self-expanding metal stents (either Magic Wallstent or Ultraflex Microvasive) and 7 children had silicone stents (2 straight, 3 Y and 2 T tube stents). At follow-up at 1 week to 72 months (median 21), only 8 of 17 (47%) children were alive but all the deaths were secondary to the underlying pathology and not related to tracheobronchial stenting. Six of 8 ventilator-dependent children were extubated after a mean of 5.3 days (range, 2 to 11) after airway stenting. For the 9 children stented for dyspnea, mean Medical Research Council dyspnea score decreased from 3.0 to 1.6 after stenting. CONCLUSIONS: Tracheobronchial stenting in children is only rarely needed and often undertaken in dire circumstances. The procedure has led to significant symptomatic benefit in dyspneic children and has enabled ventilator-dependent children to be extubated. Medium-term outlook after stenting with self-expanding metal stents for vascular compression of the airway is encouraging. The long-term outcome remains uncertain, however, and is ultimately influenced by the underlying disease. PMID- 12735583 TI - Fibrin glue in pulmonary resection: a prospective, randomized, blinded study. AB - BACKGROUND: In contrast to the rare large-airway bronchopleural fistulas after lung resection, peripheral or alveolar air leaks (AAL) are very common, often prolong hospital stay, increase utilization of resources, and on occasion result in significant morbidity. Various adjuncts have been used in attempts to reduce AAL. One of these, the topical application of fibrin glue, has to date failed to demonstrate efficacy in small clinical trials. This study reexamines the role of fibrin glue in routine lobar and wedge pulmonary resections. METHODS: Of 113 patients enrolled, 13 became ineligible because of intraoperative findings. The remaining 100 patients were randomly assigned to one of two groups at the conclusion of lung resection, regardless of the presence or absence of identifiable air leak. The control group received no additional intervention. The experimental group underwent application of 5 mL of fibrin glue delivered by a pressurized, aerosolized spraying mechanism. Postoperatively a blinded clinical observer recorded outcomes including the incidence and duration of AAL, prolonged AAL (PAAL), the volume of pleural drainage, the time to tube removal, and the postoperative length of stay (LOS), as well as any complications related to treatment. RESULTS: Both groups were comparable with regard to demographics, diagnoses, and procedures. Statistically significant reductions were found in the experimental group in the overall incidence of AAL (34% versus 68%, p = 0.001), mean duration of AAL (1.1 versus 3.1 days, p = 0.005), mean time to chest tube removal (3.5 versus 5.0 days, p = 0.02), and the incidence of PAAL (2% versus 16%, p = 0.015). There was no significant difference in the volume of chest tube drainage or LOS (4.6 days glue and 4.9 days control, p = 0.318). There were no complications related to the use of fibrin glue. CONCLUSIONS: Aerosolized fibrin glue appears to be safe and effective in reducing AAL. The overall incidence of AAL was reduced by 50% and PAAL occurred in only 1 treated patient (2% versus the usually reported 15%). Further studies with this and other methods are required to delineate routine versus selective use, to compare methods, and clarify cost benefit. PMID- 12735584 TI - Suction versus water seal after thoracoscopy for primary spontaneous pneumothorax: prospective randomized study. AB - BACKGROUND: The objectives of the study were to review our experience of video assisted thoracoscopic apical pleurectomy and to evaluate whether suction or water seal is superior in the postoperative treatment of primary spontaneous pneumothorax. METHODS: One hundred consecutive patients undergoing thoracoscopy for primary spontaneous pneumothorax from January 1995 to December 1999 were prospectively randomly assigned after surgery to receive suction or water seal to their chest tubes after a brief period of suction. RESULTS: There were 50 patients in each group. The two groups were evenly matched for age and operation performed. The method of management was stapling of an identified bleb or apex of the upper lobe and apical pleurectomy. Postoperative prolonged air leak longer than 5 days occurred in 8 patients (8%), 7 in the suction group and 1 in the water seal group (p = 0.03). The mean duration of chest tube days was lower in the water seal group (2.7 days) than in the suction group (3.8 days; p = 0.004). The mean hospital stay in the water seal group was 3.7 days and in the suction group it was 4.8 days (p = 0.004). Mean follow-up was 48 months (range, 30 to 60) for all patients. Pneumothorax recurred in 2 patients (2%). The recurrences occurred in the first year. CONCLUSIONS: Video-assisted thoracoscopic apical pleurectomy is effective and safe for treating primary spontaneous pneumothorax. Placing chest tubes on water seal after a brief period of suction shortens the duration of chest tube placement and hence the hospital stay. PMID- 12735585 TI - Loss of heterozygosity on the long arm of chromosome 21 in non-small cell lung cancer. AB - BACKGROUND: In Down syndrome, the incidence of solid tumors including lung cancer is considerably lower than that of the general population. The low risk of lung cancer in individuals with Down syndrome may be related to the gene-dosage effect of the extra chromosome 21. It may suggest that tumor suppressor genes playing a role in the pathogenesis of lung cancer may be present on chromosome 21. METHODS: A total of 39 surgically resected non-small cell lung cancers were analyzed using nine microsatellite markers for 21q. Loss of heterozygosity was considered to be present when the signal intensity of the allele in tumor DNA was less than 50% of that in the corresponding normal DNA. RESULTS: Loss of heterozygosity for at least one locus was detected in 22 of 39 tumors (56.4%). Allelic loss was frequently detected at three distinct regions: at the locus D21S1432 on 21q21.1, the region between D21S1435 and D21S1442 on 21q21.2 to 21.3, and the region between D21S1270 and D21S1445 on 21q22.1. CONCLUSIONS: These results indicate that loss of heterozygosity on 21q may play an important role in the pathogenesis of non-small cell lung cancer. PMID- 12735586 TI - Prospective study of thoracoscopic limited resection for ground-glass opacity selected by computed tomography. AB - BACKGROUND: With recent advances in low-dose helical computed tomography (CT), detection of ground-glass opacity (GGO) has increased. The aim of this study was to correlate high-resolution CT (HRCT) findings with pathologic features and to evaluate the efficacy of thoracoscopic limited resection for focal GGO, which were selected based on HRCT findings. METHODS: Focal GGO lesions were classified into two subtypes based on HRCT findings: pure type and mixed type. Ninety-six patients with persistent GGO 2 cm or less in diameter underwent pulmonary resection from January 1997 to December 2001. Among these, thoracoscopic wedge resection was performed prospectively between June 2000 and December 2001 in 33 patients with pure GGO lesions that were 1 cm or less. RESULTS: Thoracoscopic wedge resection was completed with complete safety. The histologic diagnoses of these 33 lesions were adenocarcinoma in 1, bronchioloalveolar carcinoma (BAC) in 23, and atypical adenomatous hyperplasia (AAH) in 9. No patients have had any evidence of tumor recurrence to date. Of the total 96 GGO lesions, 93.0% (53/57) of pure GGO 1 cm or less were BAC or AAH, whereas 38.5% (15/39) of pure GGO larger than 1 cm or mixed GGO were adenocarcinoma. CONCLUSIONS: Pure GGO 1 cm or less was characteristic of noninvasive lesions. Thoracoscopic limited resection for small GGO lesions selected by HRCT was valid. PMID- 12735587 TI - Videothoracoscopic management of the solitary pulmonary nodule: a single institution study on 429 cases. AB - BACKGROUND: Solitary pulmonary nodules (SPNs) sometimes are malignant. We evaluated our 9-year experience in the videothoracoscopic (VATS) management of indeterminate SPNs in order to identify malignant nodules on the basis of clinical and radiologic characteristics. METHODS: From July 1992 to May 2001, 429 patients (276 men and 153 women) were treated by VATS because of a SPN. RESULTS: Three hundred and seventy lesions were benign (86.24%): mean age, 49 years; mean diameter, 1.8 cm; evidence of calcifications, 43 cases (11.62%); smokers, 201 cases (54.32%); history of previous cancer, 11 cases (2.97%). Fifty-nine lesions (13.75%) were malignant (52 NSCLC and seven metastases): mean age, 59 years; mean diameter, 2.31 cm; evidence of calcifications, two cases (3.38%); smokers, 40 cases (67.79%); history of previous cancer, 12 cases (20.33%). Solitary pulmonary nodules were succesfully resected by VATS in 333 (77.62%) cases (309 benign and 24 malignant). Minithoracotomy was necessary to locate the SPNs in 93 (21.67%) cases (61 benign and 32 malignant). Multivariate analysis showed that age greater than 55 years (OR 4.9), diameter of the lesion greater than 2 cm (OR 4.7), history of previous cancer (OR 17.7), and need for conversion to thoracotomy (OR 6.6) to locate the nodule were statistically significant factors in order to identify malignant SPNs. CONCLUSIONS: VATS represents the approach of choice for both diagnosis and treatment of SPNs. The probability of malignancy for SPNs increases with size of the lesion, age of the patients, history of previous cancer, and the need for thoracotomy to locate the nodule. PMID- 12735588 TI - Median sternotomy for pneumonectomy in patients with pulmonary complications of tuberculosis. AB - BACKGROUND: Traditionally, a thoractomy incision is used for pulmonary complications of tuberculosis. An attractive alternative is being presented by the use of midline sternotomy in such patients, which is the aim of this study. METHODS: Five patients (four male, one female) with pulmonary complications of tuberculosis requiring surgical therapy in 1993 and 1994 were treated through a median sternotomy. The median patient age at time of surgery was 40.2 years and the median patient follow-up was 4.0 years (range 1.0 to 5.0 years) in this retrospective review. RESULTS: All patients had uncomplicated operative courses and were discharged from the hospital. One patient's in-hospital postoperative course was complicated by prolonged ventilator dependency requiring temporary tracheostomy; he died 1 year postoperatively after hospital discharge due to recurrent multidrug-resistant tuberculosis. Sternal wound infections and bronchopleural fistulas were not observed in any patients. CONCLUSIONS: Surgical treatment of pulmonary complications of tuberculosis was traditionally performed through a thoracotomy approach. Many patients with tuberculous lungs have pulmonary adhesions or intrathoracic scarring from previous surgery, which would require extrapleural resection. Bleeding was a frequent complication of this procedure. Sternotomy offers excellent exposure of the intrapericardial vessels, and reduced postoperative disability compared to the standard thoracotomy, which may be an advantage given that the majority of patients in this population have poor pulmonary function. We recommend median sternotomy as an alternative operative approach in selected patients with pulmonary complications of tuberculosis. PMID- 12735590 TI - Successful use of Argatroban as a heparin substitute during cardiopulmonary bypass: heparin-induced thrombocytopenia in a high-risk cardiac surgical patient. AB - Whereas heparin is the most widely used intravenous anticoagulant in the US for the treatment of thromboembolic disease and is a seminal adjunct to many clinical procedures, its use can cause serious adverse events. Heparin-induced thrombocytopenia (HIT) has emerged as one of the most frequently seen complications of heparin therapy and can be a life-threatening immunohematological challenge for patients requiring cardiopulmonary bypass (CPB) with obligatory heparin exposure. Unfortunately, lack of convenient monitoring techniques and the presence of HIT and other comorbidities in the complex patient frequently limits or precludes the use of most alternatives to heparin anticoagulation during CPB. This case report describes the successful use of the celite activated clotting time and high-dose thrombin time, while using the direct thrombin inhibitor Argatroban as an alternative to heparin anticoagulation during CPB in a high-risk patient presenting with type II HIT, end-stage renal failure, and ischemic cardiomyopathy with ventricular fibrillatory arrest. PMID- 12735589 TI - Lower sternal reinforcement improves the stability of sternal closure. AB - BACKGROUND: This study uses a mechanical testing system to evaluate three methods of sternal closure. METHODS: Twelve sternal replicas composed of a polyurethane foam bone analogue were divided in the midline and reapproximated using three stainless steel wire techniques: six simple wires (6S), six figure-of-eight wires (6F8), or seven simple wires (7S), which included an extra wire at the lower sternum. The closures were subjected to increasing lateral distraction from 0 to 400 Newtons (N) (1 N = 0.224 lbs), and motion was measured using transducers stationed across the manubrium, midsternum, and lower sternum. RESULTS: With each method of closure, the manubrium was the most stable, the lower sternum the least stable, and the midsternum intermediate between the other two. There were also differences between sternal closure methods, but only at the lower sternum. Less sternal distraction was measured with the 7S than the 6S and 6F8 methods, starting at 100 N (0.20 +/- 0.06 mm vs 0.48 +/- 0.19 and 0.39 +/- 0.10, p = 0.003), and progressively increasing until the study was stopped at 400 N (1.64 +/- 0.39 mm vs 4.92 +/- 1.73 and 5.1 +/- 1.43 mm, p = 0.003). CONCLUSIONS: These data show that the lower sternum is the site of greatest instability and that reinforcement of this area with an additional wire effectively stabilizes the closure. Figure-of-eight wires are not superior to simple wires. PMID- 12735591 TI - Timing of transesophageal echocardiography in diagnosing patent foramen ovale in patients supported with left ventricular assist device. AB - Left ventricular assist devices unload the left ventricle and decrease left atrial pressure. This hemodynamic change may cause a right to left atrial shunt and hypoxemia in patients with patent foramen ovale. We prospectively studied the best time for performing diagnostic transesophageal echocardiography in left ventricular assist device patients. Intraoperative transesophageal echocardiography was performed in 14 patients before cardiopulmonary bypass was initiated and after left ventricular assist device was implanted. No patent foramen ovale was detected when transesophageal echocardiography was done before bypass, but a patent foramen ovale was found in 3 patients when transesophageal echocardiography was performed after left ventricular assist device was activated. Patent foramen ovale was confirmed by inspection in all three patients and surgically closed during the same procedure. There were no patent foramen ovale closure-related complications. PMID- 12735593 TI - Chronic expanding hematoma in the pericardial cavity after cardiac surgery. AB - We report the successful surgical treatment of a rare case of chronic expanding hematoma in the pericardial cavity that developed into a very large mass over a long period. The patient, who had a history of cardiac surgery for a double outlet right ventricle 14 years previously and for tricuspid regurgitation 8 years ago, noticed a slowly growing mass near the left atrium and ventricle 5 years ago. The mass, which confirmed a diagnosis of chronic expanding hematoma, was resected by left thoracotomy. PMID- 12735592 TI - Graft occlusion after deployment of the Symmetry Bypass System. AB - Recently the Symmetry Bypass System (SJM, St. Paul, MN) became available. Now the system is frequently applied for vein-graft to aorta anastomoses in off-pump coronary artery bypass operations. This report describes a complication associated with the use of the Symmetry Bypass System (SJM) in a patient undergoing a standard off-pump coronary artery bypass procedure. A novel imaging system (SPY, Novadaq, Toronto, Canada) was applied for intraoperative assessment of graft function, and this system immediately diagnosed the occlusion of the proximal mechanical anastomosis caused by a mobile atheromatous aortic plaque. PMID- 12735594 TI - Detection of pannus by multidetector-row computed tomography. AB - Prosthetic valve dysfunction (PVD) due to pannus formation is an infrequent but serious complication. A 72-year-old man who underwent aortic valve replacement was diagnosed with PVD and aneurysm of the Sinus of Valsalva. Multidetector-row computed tomography (MDCT) was used to examine the cause of PVD before reoperation. MDCT demonstrated that tissue regarded as pannus extended from the left ventricular septum into the pivot guard. These findings were confirmed by observations during reoperation. MDCT can be a useful diagnostic technique for the anatomical and functional evaluation of PVD. PMID- 12735595 TI - Unexpected durability of Smeloff-Cutter aortic ball valve prosthesis. AB - We report a case in which replacement of a Smeloff-Cutter aortic ball prosthesis was required 28 years after initial implantation. A 57-year-old woman underwent aortic valve replacement with a 21-mm Smeloff-Cutter ball prosthesis and open mitral commissurotomy for aortic stenosis, aortic regurgitation, and mitral stenosis in 1973. Severe aortic regurgitation occurred in April 2001, and aortic valve reoperation combined with mitral valve replacement was successfully performed. The patient's aortic ball valve was nearly intact with perivalvular leakage probably causing the aortic regurgitation. Our experience documents longer durability for the Smeloff-Cutter prosthesis than has been reported to date. PMID- 12735596 TI - Staged laparoscopic splenectomy and valve replacement in splenic abscess and infective endocarditis. AB - Splenic abscess is a rare clinical entity that is most commonly associated with infective endocarditis. Valve replacement in the setting of an unaddressed splenic abscess is associated with a high incidence of prosthetic valve infection and death. We describe 2 patients with infective endocarditis and splenic abscess treated by laparoscopic splenectomy followed by valve replacement. PMID- 12735597 TI - Traumatic arteriovenous fistula of ascending aorta and left innominate vein. AB - We report a patient with a traumatic arteriovenous fistula between the ascending aorta and innominate vein, presenting 4 years after injury. Diagnostic imaging and surgical management of this rare manifestation of thoracic great vessel injury is described. PMID- 12735598 TI - New approach to the surgical management of pulmonary arteriovenous malformations after cavopulmonary anastomosis. AB - The development of pulmonary arteriovenous malformations after cavopulmonary bypass in patients with congenital heart disease is well documented. We report successful management of pulmonary arteriovenous malformations after cavopulmonary bypass in a patient with an interrupted inferior vena cava (IVC) and multiple hepatic veins utilizing an extracardiac conduit from the hepatic veins to the hemiazygous continuation of the interrupted IVC. This technique, performed without circulatory arrest or an atriotomy, may limit morbidity associated with intracardiac procedures in patients with single ventricle morphology. Furthermore, this case suggests an alternative technique for completion Fontan in patients with an interrupted IVC and multiple hepatic venous drainage. PMID- 12735599 TI - Surgical treatment of the left circumflex coronary artery from the pulmonary artery in an adult patient. AB - We report an exceptional case of ischemic heart disease due to the origin of the left coronary circumflex artery from the pulmonary artery in a 50-year-old woman. She had undergone surgery for aortic coarctation when she was 16 years old. This abnormality was associated with other congenital defects such as tunnel subaortic stenosis, small aortic valve annulus, numerous left ventricular false tendons, and aortic bicuspid valve. Cardiac surgery verified the origin of the left circumflex from the pulmonary artery. The left internal mammary artery was positioned on the obtuse marginal coronary branch. Her clinical state was moderately improved 3 months after surgery. PMID- 12735600 TI - Intrapleural tumor dissemination after video-assisted thoracoscopic surgery metastasectomy. AB - In a 34-year-old woman extensive intrapleural disease developed within 1 year of video-assisted thoracoscopic surgery (VATS) removal of two pulmonary metastases from a previously treated synovial sarcoma. She underwent a successful complete pleurectomy through a left thoracotomy to excise the pleural tumor and remains well 6 months later. This case highlights tumor dissemination as one of the pitfalls of VATS metastasectomy and raises concerns about using VATS in this situation. PMID- 12735601 TI - Malignant "Triton" tumor of the posterior mediastinum: prolonged survival after staged resection. AB - Malignant "Triton" tumor is a rare subtype of malignant peripheral nerve sheath tumor showing rhabdomyoblastic differentiation. We report the case of a 22-year old patient who had excision of such a tumor by video-assisted thoracic surgery. He was reoperated on by thoracotomy 2 months later and received adjuvant radiation, with prolonged survival. Radical surgical excision of malignant Triton tumor followed by adjuvant radiation therapy may provide the longest survival and represent the treatment of choice. PMID- 12735602 TI - Castleman's disease associated with myasthenia gravis. AB - Castleman's disease presents as a peculiar type of lymph node hyperplasia. Traditionally, the disease has been classified on clinical grounds (solitary or multicentric) and by histologic appearance (hyaline vascular pattern, plasma cell predominance, or mixed lesions). It is now increasingly clear that there are different etiologies for each of these different subtypes. Reported associations include POEMS syndrome (polyneuropathy, organomegally, endocrinopathy, monoclonal gammopathy, and skin changes), paraneoplastic pemphigus, Hodgkin's disease, and follicular dendritic cell sarcoma. We present a case of Castleman's disease associated with myasthenia gravis, the third reported case in the literature. We discuss Castleman's disease and review the literature. PMID- 12735603 TI - Chylotamponade: an unusual presentation of Gorham's syndrome. AB - Gorham's Syndrome, also known as massive osteolysis or "vanishing bone disease" results from lymphangiomatosis with adjacent bone resorption. Chylothorax is a common complication in cases of mediastinal involvement. We report a case of Gorham's Syndrome presenting as chylotamponade successfully treated with pericardial drainage, early parenteral nutritional support, bilateral pleurodesis for chylous effusions, and adjuvant external beam radiation. PMID- 12735604 TI - Cystic pulmonary metastases from epithelioid cell sarcoma. AB - A patient with cystic pulmonary metastases, from epithelioid sarcoma of the thigh, resulting in bilateral pneumothoraces is presented. The pulmonary metastases superficially resembled pneumomatoceles and were not visible on radiography. Computed tomography examinations over a 10-month period showed no change in the size or number of the cystic metastases, though air fluid levels and pericystic nodular consolidation (probably due to pulmonary hemorrhage) did occur and regress. This case report serves to emphasize that bullous type lesions on thoracic computed tomography in patients with a known sarcoma should be interpreted with particular caution. PMID- 12735605 TI - Thoracotomy for correction of a kinked right internal mammary artery graft. PMID- 12735606 TI - Pulmonary invasion in multisystem Langerhans cell histiocytosis. PMID- 12735607 TI - A new method for reliable fenestration in extracardiac conduit Fontan operations. AB - A fenestrated extracardiac conduit Fontan operation was performed with a new method in 5 patients by means of a pericardial tube anastomosed end to end with the inferior inlet of the right atrium. PMID- 12735608 TI - A method for descending thoracic aortic replacement retaining a posterior strip bearing intercostal vessels. AB - Operations for aneurysms of the descending thoracic aorta are still fraught with danger. Spinal cord injury remains a major cause of morbidity. Many therapeutic strategies have been suggested to reduce the incidence of this devastating complication, including reimplantation of intercostal vessels. However, reimplantation of intercostal vessels, both individually or in groups, is time consuming and compounded by the absence of a reliable means of identifying which vessels actually supply the cord. We present a technique that allowed inclusion of all potentially important descending aortic branching vessels into the repair leading to a favorable outcome in a series of patients. PMID- 12735609 TI - A new method for detection of postpneumonectomy broncho-pleural fistulas. AB - Broncho-pleural fistula is a relatively rare but life-threatening complication of pneumonectomy. Early detection of this complication is crucial for optimal treatment. We describe a simple technique to detect postpneumonectomy broncho pleural fistula by measuring concentrations of O2 and N2O in the pneumonectomy cavity at baseline and after allowing patients to breath gas mixtures enriched with O2, N2O, or both. The technique was used in 22 patients. In 20 control patients the test was carried out 48 hours after pneumonectomy. Both the O2 and the N2O test were negative in all cases. However, in 2 patients with broncho pleural fistula both the O2 and the N2O tests were positive. PMID- 12735610 TI - Hand-assisted thoracoscopic surgery. AB - Hand-assisted thoracoscopic surgery is a novel minimally invasive approach for performing techniques conventionally performed by posterolateral thoracotomy. Hand-assisted thoracoscopic surgery overcomes one of the major drawbacks of minimally invasive thoracic surgery in allowing full manual palpation of the lungs through a subcostal incision under video guidance, while avoiding a thoracotomy when the indication is pulmonary metastasectomy with curative intent or resection of undiagnosed lung nodules. The technique may result in improved quality of life outcomes compared with a thoracotomy. PMID- 12735611 TI - Cardioplegia in pediatric cardiac surgery: do we believe in magic? AB - Cardioplegia has become the gold standard of myocardial protection for practically every type of heart surgery during which the ascending aorta must be clamped. Although there is little doubt about the efficacy of cardioplegia in the adult heart, there are few studies on the pediatric heart and their results are contradictory. The physiology of pediatric heart muscle differs considerably from that of the adult myocardium. The pediatric heart distinguishes itself from that of the adult most impressively in its greater tolerance for ischemia. This ischemia tolerance is enhanced by the use of hypothermia. Considering that hypothermia is a powerful tool to prolong ischemia tolerance and that most pediatric cardiac surgeons report similar results using different types of cardioplegia, some surgeons are tempted to suspect that the contribution of the cardioplegia composition to protecting the pediatric heart may be overestimated. This provocative statement is critically discussed in this article. We examine the protective potential of cardioplegia (in various compositions), or of hypothermia, or of both in pediatric cardiac surgery. We pay special attention to several key differences between the physiologies of the pediatric myocardium and the adult myocardium and attempt to relate them to the available surgical methods of myocardial protection. We conclude that the composition of cardioplegia indeed is an important component of successful operative management in pediatric heart surgery. We provide evidence that the benefit of cardioplegia over hypothermia alone is minor at low temperatures (below 15 degrees C), but becomes substantial when the temperature increases. PMID- 12735612 TI - Heart valve choices in the adult: mechanical or bioprosthetic? PMID- 12735613 TI - Pulmonary vein isolation and left atrial reduction for chronic atrial fibrillation secondary to mitral valve disease. PMID- 12735614 TI - Glove punctures in cardiac surgery. PMID- 12735615 TI - Skeletonization of arterial conduits: method of choice. PMID- 12735616 TI - Does hydroxyethyl starch increase blood loss in cardiac operations. PMID- 12735617 TI - A suggestion regarding catamenial pneumothorax. PMID- 12735618 TI - Intramedullary stabilization of distal-third femur fractures in octogenarians. AB - Management of distal-third femoral fractures is challenging, especially in the elderly population, where the associated complication rate has been reported to be as high as 40%. This study reviews the techniques and safety of intramedullary stabilization for displaced and unstable fractures involving the distal third of the femur in patients who were at least 80 years old, with the use of Ender nails. A retrospective study of 12 patients with 14 femur fractures underwent closed fixation of their fracture(s) with Ender nails performed within 24 hours of the injury. There were 10 females and 2 males, whose average age was 88 years (range, 82-103 years). The average length of follow-up was 3 years. Twelve fractures (86%) healed uneventfully. There were two nonunions (14%) with no other major or minor complications. No patients were lost to follow-up evaluation. All patients eventually returned to their preinjury level of function. This technique is particularly applicable to elderly patients with severe osteopenia and preexisting medical problems, as it minimizes blood loss and other associated complications, while providing stability for immediate mobilization. PMID- 12735619 TI - Postoperative deep vein thrombosis prophylaxis: a retrospective analysis in 1000 consecutive hip fracture patients treated in a community hospital setting. AB - The occurrence of deep vein thrombosis (DVT) following cases of major trauma, in particular pelvic and hip fracture, has ranged from 36% to 60%, depending on the study quoted and the method of detection. The frequency of fatal pulmonary embolism (PE) has been reported as 0.5%-12.9% of the cases. A retrospective study of 1000 consecutive hip fracture patients in a community hospital setting reveals that 95% received a combination of mechanical and pharmacologic prophylaxis for prevention of DVT. Sixty-one patients were excluded for insufficient data, leaving 939 for analysis. There were 724 female patients with an average age of 83 years and 215 male patients with an average age of 78 years. Fifty-one patients (18.4%) received no prophylaxis in the eligible population. Three hundred eighty-seven patients (41.2%) received only aspirin as the pharmacologic agent for anticoagulation. Four hundred twenty-nine patients (45.6%) were treated with the low-molecular-weight heparin (LMWH), enoxaparin. Ten patients (1.1%) received heparin for anticoagulation and 17 patients (1.8%) were treated with warfarin. A total of 43 patients received a combination of therapies. Four hundred ninety-five of the patients used concomitant intermittent pneumatic compression in addition to pharmacologic prophylaxis. There were 15 perioperative deaths from all causes, including five cases of DVT two distal and three proximal). One distal DVT occurred prior to surgery. A second distal DVT and one fatal PE occurred in the aspirin group. The rates of minor bleeding complications in the aspirin group, the < 12-hour postoperative dosing of the enoxaparin group, and the 12 to 24-hour postoperative dosing of the enoxaparin group were 3.1%, 5.7%, and 2.8%, respectively. There were no major bleeds in the aspirin group and 0.9% in the enoxaparin group. The LMWH group also had two proximal DVTs but no PEs. The combination of a relatively short half-life, predictable pharmacokinetics, and favorable safety profile makes enoxaparin an excellent drug for use in hip fracture patients. Additional trials will be necessary to establish an optimal duration of prophylaxis in this population. PMID- 12735620 TI - Unrecognized risks among Veterans with hip fractures: opportunities for improvements. AB - Hip fractures are associated with significant morbidity and mortality. Most of these fractures are preventable late manifestations of osteoporosis. The authors examined incidence and medical costs associated with hip fractures among 34,363 veterans by gender and race for fiscal years 1996-2000. Data were obtained from the Patient Treatment File administrative database. The frequencies of seven known risk factors were quantified. White veterans hospitalized for hip fracture were 69% more likely to be given the diagnosis of osteoporosis, when compared to black veterans. Male gender was associated with lower percentage of osteoporosis diagnoses (2.2%) than female gender (11.9%). The authors estimate that hip fractures result in 43 million dollars of excess cost to veterans and the Veterans Health Administration annually. Greater use of diagnostic and preventive measures represents an excellent opportunity for improvement of care, as well as substantial cost savings for at risk populations. PMID- 12735621 TI - Correlation of age, acromial morphology, and rotator cuff tear pathology diagnosed by ultrasound in asymptomatic patients. AB - The importance of acromial morphology in the pathogenesis of rotator cuff tears remains controversial. Some surgeons place great emphasis on acromial morphology and others feel that acromial shape is a result of cuff tear pathology rather than the cause. The purpose of this study was to determine if there was an association between acromial morphology, age, and rotator cuff tears found by ultrasound in asymptomatic volunteer subjects with no past history of shoulder symptoms. One hundred eighteen outlet x-rays and ultrasound scans were performed in 59 asymptomatic patients in various age groups. Acromial morphology and the age of the patients were then correlated with the ultrasound findings. Older patients were noted to have a high incidence of type II and type III acromions (93% of those over 70). Full- and partial-thickness tears were more commonly seen in patients with type II or type III acromions compared with type I. Patients over 50 years had a high incidence of full-thickness tears (40%), but the incidence did not increase with increasing age past 50. These findings lend credence to the multifactorial etiology of rotator cuff tears. The patient's age (degeneration) and acromial morphology (impingement) are two of the factors involved in causing rotator cuff tears. The incidence of cuff tears and type III acromions was high in this group of entirely asymptomatic volunteer subjects. Surgeons should interpret radiologically hooked or curved acromions as well as rotator cuff tears diagnosed with ultrasound or other modalities with caution. It may well be that both of these findings should be regarded as part of the natural ageing process. PMID- 12735622 TI - Concomitant nonunion of the distal humerus and olecranon. AB - Concomitant nonunion of an operatively treated fracture of the distal humerus and the olecranon osteotomy used for exposure is an unusual and complex situation which compromises ulnohumeral function on both sides of the joint. Operative treatment restored good elbow function in a series of six patients with this problem. An average of 110 degrees of ulnohumeral motion was restored, arthrosis was none (four patients) or mild (two patients) at an average follow-up of 50 months, and outcome measures documented good upper extremity specific (DASH) and general (SF-36) health status. PMID- 12735623 TI - Early acetabular protrusio following hemiresurfacing of the hip for osteonecrosis in sickle cell disease. AB - A case report is presented of a patient with acetabular protrusio 2 years following hemiresurfacing of the femoral head for osteonecrosis associated with sickle cell disease. Renewed interest in bone-preserving hip arthroplasty has led to increased use of hemiarthroplasty and femoral resurfacing in young patients with arthritis. In cases of osteonecrosis, especially that associated with sickle cell disease, awareness of this potential complication is important. PMID- 12735624 TI - Anterior inferior iliac spine apophyseal avulsion fracture. AB - A case of anterior inferior iliac spine (AIIS) apophyseal avulsion fracture caused while playing football is reported. A 16-year-old amateur football player felt severe pain in his left groin while kicking the ball during training. There was point tenderness over the anterior inferior iliac spine (AIIS). Avulsion fracture of AIIS was considered clinically. Radiographs confirmed the diagnosis. He was treated with a conservative rehabilitation program. He is still an active football player. Avulsion fractures follow violent or explosive muscular contractions against a fixed resistance, sudden deceleration, or stretching of the involved muscle or as a result of a direct trauma. This injury usually occurs with an extension moment to the hip joint, with the knee flexed, and it is commonly seen in sports that involve kicking. AIIS avulsion fractures should be a diagnostic consideration in patients with pain in the groin who are involved in activities requiring high-level forces of flexion of the hip. PMID- 12735625 TI - High ulnar nerve compression by a triceps branch schwannoma. AB - An unusual case is presented in which a major peripheral nerve was compressed extrinsically by a schwannoma derived from a minor en passant triceps nerve branch. PMID- 12735626 TI - A new-generation apparatus for studying memory-related performance in mice. AB - 1. We developed a new kind of food search test that can measure murine nocturnal memory without handling hard work for setting up. 2. This apparatus has four food stations, but only one station had accessible food at any time. The one station with accessible food was changed at 4-h intervals. 3. We compared the performance of transient forebrain global Ischemic mice, which are a hippocampal lesion model, with the performance of control C57BL/6J mice. 4. The correct visit ratio, i.e., the ratio of the number of visits to the correct food station to the number of visits to all stations, gradually increased in the control mice, but did not change in the Ischemic mice. 5. This new system was demonstrated to be an additional and useful tool for studying memory-related performance in mice. PMID- 12735627 TI - The effect of an endogenous Na+, K+-ATPase inhibitor on rat lens transparency and ultrastructure. AB - 1. The purpose of the present study was to analyze the possible effect of ouabain and an endogenous ouabain-like substance (endobain E), on lenses of 100- and 400 g body weight rats. 2. Lenses were incubated with ouabain or endobain E for 120 min, either at room temperature or in the cold; opalescence was checked by gross examination and ultrastructure by electron microscopy. 3. Lenses from 400-g rats invariably remained translucent whereas those from 100-g rats presented variable opalescence. 4. As disclosed with the electron microscope, lenses of 100-g rats incubated at room temperature, with or without ouabain or endobain E, presented variable degrees of ultrastructural changes: with ouabain, there was fiber separation and vacuole formation but with endobain E, no vacuoles were found and fibers, though disorganized, appeared attached. After incubation in an ice bath, lenses were markedly altered in all conditions assayed. 5. It is concluded that ouabain and endobain E effect on lens transparency depends on the rat age and that in young animals, it is crucial incubation temperature during experimental procedure. PMID- 12735629 TI - NR2A but not NR2B N-methyl-D-aspartate receptor subunit is altered in the visual cortex of BDNF-knock-out mice. AB - 1. Aim of the present paper is to study the expression of N-Methyl-D-Aspartate receptor (NMDAR) subunits NR2A and NR2B within mouse visual cortex. 2. To investigate the influence of neurotrophic factor of NGF family (neurotrophins) on NMDAR expression we used mutant mice carrying a deletion in the gene for brain derived neurotrophic factor (BDNF), a well-known neurotrophin expressed in visual cortex. 3. Western blot and immunohistochemistry were performed at postnatal day P12-14, P21-23, and adulthood showing that both subunits change during postnatal development. 4. Absence of BDNF induced a reduction of NR2A level. This effect was specific since the other subunit investigated, NR2B, was not affected in mutant mice. 5. We conclude that endogenous BDNF modulates NMDAR expression in the developing visual cortex. PMID- 12735628 TI - Early postnatal sound exposure induces lasting neuronal changes in the inferior colliculus of senescence accelerated mice (SAMP8): a morphometric study on GABAergic neurons and NMDA expression. AB - Senescence-acceleration-prone mice (SAMP8) provide a model to study the influence of early postnatal sound exposure upon the aging auditory midbrain. SAMP8 were exposed to a 9-kHz monotone of either 53- or 65-dB sound pressure level during the first 30 postnatal days, the neurons in the auditory midbrain responding selectively to 9 kHz were localized by c-fos immunohistochemistry and the following parameters were compared to control SAMP8 not exposed to sound: mortality after sound exposure, dendritic spine density, and quantitative neurochemical alterations in this 9-kHz isofrequency lamina. For morphometric analysis, animals were examined at 1, 4, and 8 months of age. Serial sections of the inferior colliculus were Golgi impregnated or stained immunohistochemically for the expression of epsilon1 subunit of NMDA receptor or GABA. Mortality after exposure to 53 dB was the same as in controls, but was markedly increased from 7 months of age onward after postnatal exposure to 65 dB. No gross morphological alterations were observed in the auditory midbrain after sound exposure. However, sound exposure to 53 or 65 dB significantly reduced dendritic spine density by 11% at 4 months or by 11-17% both at 1 and 4 months of age, respectively. The effect of sound exposure upon neurons expressing the NMDAepsilon1 subunit was dose-dependent. Increasing with age until 4 months in control mice and remaining essentially stable thereafter, the percentage of NMDAepsilon1-immunoreactive neurons was significantly elevated by 40-66% in 1- and 8-month-old SAMP8 exposed to 53 dB, whereas no significant effect of 65 dB was apparent. The proportion of GABAergic cells declined with age in controls. It was significantly decreased at 1 month after 53 and 65 dB sound exposure. In contrast, it was elevated at later stages, being significantly increased at 4 months after exposure to 53 dB and at 8 months after exposure to 65 dB. The total cell number in the 9-kHz isofrequency lamina of SAMP8 decreased with age, but was not affected by exposure to either 53 or 65 dB. The present results indicate that early postnatal exposure to a monotone of mild intensity has long-term effects upon the aging auditory brain stem. Some of the changes induced by sound exposure, e.g., decline in spine density, are interpreted as accelerations of the normal aging process, whereas other effects, e.g., increased NMDAepsilon1 expression after 53 dB and elevated GABA expression after both 53 and 65 dB, are not merely explicable by accelerated aging. PMID- 12735630 TI - Intrathecal adenosine A1 receptor agonist attenuates hyperalgesia without inhibiting spinal glutamate release in the rat. AB - The analgesia effects of intrathecal adenosine A1 receptor agonist, R-PIA, on the hyperalgesia and CSF-glutamate release after formalin injection into the rat paw were evaluated. R-PIA significantly and dose-dependently attenuated increases in flinching behavior, and this attenuating effect was reversed by the adenosine A1 receptor antagonist, aminophylline. Morphine blocked flinchs, however MK-801 partially abolished. The increase in CSF-glutamate release evoked by formalin stimulation was inhibited by morphine but not by either R-PIA or MK-801. These findings suggest that the intrathecal adenosine A1 receptor agonist provokes analgesic effect via the postsynaptic action independent of an effect upon spinal glutamate release. PMID- 12735633 TI - Production of retroviral vectors in primary human keratinocytes after DNA mediated gene transfer leads to prolonged gene expression. AB - Prolonged stability and controlled expression of gene constructs transferred directly to human skin improve the possibility of using this tissue in somatic gene therapy. We aim to develop a simple transfection method resulting in retroviral mediated gene transfer to keratinocyte stem cells in situ. We here show that after DNA-mediated gene transfer into primary human keratinocytes it is possible to achieve production of retroviral vectors, leading to the transduction of co-cultured keratinocytes and prolonged reporter gene expression. The method is a first step in a strategy to generate retroviral producer cells in situ in the skin furthermore the method can be used for rapid analysis of the possible effects of transgenes in cultured human keratinocytes without preparatory retroviral vector production in packaging cell lines. PMID- 12735631 TI - The mechanism of presynaptic long-term depression mediated by group I metabotropic glutamate receptors. AB - 1. Metabotropic glutamate receptors (mGluRs) are known to play a role in synaptic plasticity. In a study of rat hippocampal brain slices, we find that a brief perfusion of a group I mGluR agonist, (S)-3,5-dihydroxyphenylglycine (DHPG), induced a robust long-term depression (DHPG-LTD) in area CA1. 2. The action was accompanied by an enhancement of the paired-pulse facilitation (PPF) ratio. 3. At the same time DHPG enhanced ionophoretic responses to alpha-amino-3-hydroxy-5 methyl-4-isoxazole-propionic acid (AMPA), kainic acid (KA), and N-methyl-D aspartate (NMDA) in CA1 pyramidal neurons. This was only partially reversed by washing. 4. These observations indicate that DHPG exerts two opposing actions, suppression of the synaptic transmission and facilitation of postsynaptic responses. However, the presynaptic action dominates, since the net effect of monosynaptic activation is a reduction of response. 5. Perfusion of DHPG reduced three calcium-dependent responses in CA3 pyramidal neurons, which are presynaptic to CA1 neurons. These are calcium spike width and amplitude, after hyperpolarization (AHP), and spike frequency adaptation (SFA). 6. These results suggest that the DHPG-LTD results from modulation of the presynaptic calcium currents by group I mGluRs. PMID- 12735634 TI - The vulva skin microclimate: influence of panty liners on temperature, humidity and pH. AB - Many women use panty liners between menstrual periods. The aim of this study was to investigate whether the use of such products might influence the vulva skin. Twelve healthy women were studied on four occasions with three different product constructions and on one occasion without products. Temperature, surface wetness and surface pH were measured on vulva skin. Mean skin temperature when the women were wearing a conventional panty liner (with a non-breathable back sheet) was 35.9 degrees C, compared to 34.4 degrees C when wearing no panty liner at all (p < 0.01) and 34.5 degrees C when using a panty liner with a breathable (i.e. vapour permeable) back sheet (p < 0.01). Skin humidity was significantly higher when the conventional panty liner was used compared to no panty liner or to the breathable panty liner (both cases p < 0.01). The mean pH value at the exterior aspect of the labium majus was 5.8 with the conventional panty liner, 5.2 with no panty liner and 5.3 with the breathable panty liner (p < 0.001 and p < 0.01, respectively). The results indicate that the conventional panty liner changes the vulva skin microclimate, but that the breathable panty liner to a substantial degree keeps the microclimate at an undisturbed level. The actual effect of these differences on microbiological flora will be addressed in a subsequent study. PMID- 12735632 TI - A novel method of eliminating non-neuronal proliferating cells from cultures of mouse dorsal root ganglia. AB - 1. We hypothesized that non-neuronal cells could be eliminated from primary dorsal root ganglion (DRG) cultures by including a DNA topoisomerase inhibitor (camptothecin) during culture. 2. Exposure to 20 microM camptothecin for 48 h, beginning at 3 days in vitro, reliably eliminates proliferating non-neuronal cells. 3. Following camptothecin treatment, neurons survived and continued to extend neurites for several weeks without obvious defects in morphology or viability. 4. Transient camptothecin exposure is therefore an efficient and fast acting method to purify DRG neurons in culture. PMID- 12735636 TI - Expression of p53 in normal sun-exposed and protected skin (type IV-V) in different decades of age. AB - The checkpoint protein p53, which is activated by DNA damage, is involved in the decision whether the cells should stop replication and proceed to repair their DNA or die by apoptosis. We evaluate the expression of p53 and the number of apoptotic cells in normal sun-exposed (face) and protected (abdomen) skin in Egyptians between 6 and 77 years of age. The degree of p53 expression in facial skin significantly increases from a score of 1.5 +/- 1.5 (mean +/- SEM) in the 1st decade to 4.8 +/- 0.3 in the 8th decade (p = 0.02), while no significant changes are detected in the protected skin (p = 0.1). Overall, the level of expression is significantly higher in sun-exposed facial skin than in abdominal skin (p = 0.007). However, p53 expression versus age is significantly higher in the facial skin of older age groups in both males (p = 0.003) and females (p = 0.02). The pattern of staining was found to be dispersed (wild-type) in the majority (97.3%) of biopsies from sun-exposed skin and in all biopsies from non exposed skin. The expression of wild-type p53 in type IV-V skin therefore correlates with both site and age of the individual. In contrast, the number of apoptotic cells significantly decreases with advancing age in sun-exposed skin (p = 0.005). Increased age-associated expression of p53 in sun-exposed skin, but not in protected areas of skin, is found to reflect an accumulation of the wild-type protein, as judged by the staining pattern. The decrease in apoptotic cells with age may suggest the accumulation of senescent cells in the skin and their relative resistance to apoptosis. Such alteration in the proliferation/apoptosis balance could play a role in tumorigenesis. PMID- 12735635 TI - Effect of a moisturizer on skin susceptibility to NiCl2. AB - The aim of this study was to evaluate whether a moisturizer used on normal skin can increase skin response to allergens. Twelve nickel-allergic volunteers applied a lipid-rich moisturizer on the upper arm 3 times daily for 7 days, while the other upper arm served as a control. A control group followed the same treatment protocol. Following treatment with moisturizer, patch tests with 1% NiCl2 aqueous solution were applied on each upper arm. After 24 and 72 h, skin reactions were evaluated blinded by clinical scoring, and by bioengineering methods measuring transepidermal water loss, skin colour and skin thickness. In the nickel-allergic group the strength of patch-test reactions was increased on the moisturizer-treated arm as evaluated by clinical scoring after 24 h and by measurement of transepidermal water loss and skin thickness after 72 h. In the control group, no significant differences were found. Our findings show that threshold values for elicitation of allergic reactions in already sensitized individuals may be influenced by use of lipid-rich moisturizers. PMID- 12735637 TI - Matrix metalloproteinases-2 and -9 are secreted from human fibroblasts. AB - To investigate the pathophysiological role of fibroblasts in tissue metabolism, we analysed gelatin-degrading enzymes secreted from skin cells in culture. Using a gelatin-zymography method, matrix metalloproteinase (MMP)-2 was constitutively secreted from fibroblasts, whereas both MMP-2 and MMP-9 were secreted from keratinocytes. MMP-9 expression by fibroblasts was induced by the addition of transforming growth factor (TGF)-beta1 or tumour necrosis factor (TNF)-alpha to the cultures. Moreover, TGF-beta1, TNF-alpha, or epidermal growth factor augmented the secretion of MMP-2 from fibroblasts. These results indicate that MMP-9 together with MMP-2 secreted from fibroblasts could play important roles in tissue metabolism, including cytokine-induced inflammation. PMID- 12735638 TI - Matrix metalloproteinase-19 is expressed by keratinocytes in psoriasis. AB - Keratinocyte hyperproliferation, inflammatory infiltrates, neoangiogenesis and alterations in cytokine levels are hallmarks of psoriatic skin. Matrix metalloproteinases (MMPs) have been associated with the remodeling of the extracellular matrix during inflammation, neovascularization, and malignant transformation. We have previously shown that particularly MMP-12 is abundantly expressed by macrophages and MMP-9 in macrophages and neutrophils of psoriatic lesions. In this work the expression of two novel metalloproteinases, MMP-19 and MMP-28, was investigated in psoriatic lesional and non-lesional skin. MMP-19 protein was detected by immunohistochemistry in 28/29 samples in keratinocytes in the same regions as Ki67 (marker of proliferating keratinocytes) and p63 (marker of keratinocyte stem cells). Immunosignaling was also seen in endothelial cells and fibroblasts. Furthermore, MMP-19 mRNA was upregulated in psoriatic keratinocytes and skin as assessed by quantitative real-time polymerase chain reaction. In lichen planus and lichenoid chronic dermatitis, MMP-19 staining was found in keratinocytes in areas where the basement membrane was abnormal. MMP-28 was not detected in psoriatic or non-lesional skin. Our results suggest that keratinocytes as well as the previously reported cell types (smooth muscle, endothelial and macrophages) can express MMP-19 in psoriasis and lichen planus. Upregulation of MMP-19 in keratinocytes may be influenced by changes in the architecture of the basement membrane zone. PMID- 12735639 TI - Familial and sporadic porphyria cutanea tarda: clinical, biochemical and genetic features with emphasis on iron status. AB - The manifestation of porphyria cutanea tarda reflects genetic and environmental factors. Mutations in the uroporphyrinogen decarboxylase gene, located at chromosome 1p34, discriminate familial porphyria cutanea tarda from sporadic cases. Furthermore, mutations in the haemochromatosis gene may be involved in the aetiology. In this study 53 unrelated Danish patients with porphyria cutanea tarda were classified according to uroporphyrinogen decarboxylase and haemochromatosis gene mutations and the genotype related to the clinical and biochemical data. Thirteen patients (25%) had familial porphyria cutanea tarda. The results signify the advantage of DNA diagnostics for identification of familial cases, as anamnestic data are doubtful and erythrocyte uroporphyrinogen decarboxylase activity measurements insufficient for correct classification. Eight patients with porphyria cutanea tarda (15%) were homozygous for the haemochromatosis gene C282Y mutation and 8 patients were heterozygous. Patients homozygous for the haemochromatosis related mutation showed biochemical evidence of excessive iron storage as well as increased urine porphyrin excretion levels. This seems to confirm a relationship between porphyria cutanea tarda and haemochromatosis. No differences were found between patients with sporadic and familial porphyria cutanea tarda regarding age of onset, clinical severity, sex distribution, liver function tests and iron storage parameters. However, daily alcohol intake and use of oestrogens were reported more frequently in the group of sporadic patients. It was found that women were over-represented in our study. PMID- 12735640 TI - Microorganisms in intertriginous psoriasis: no evidence of Candida. AB - Infection can be a trigger and an aggravating factor in psoriasis. Antibacterial and/or antifungal agents are commonly used in the treatment of intertriginous psoriasis, because it is believed that flexures in psoriasis are often colonized by Candida species and Staphylococcus aureus. Bacterial and fungal cultures were studied from 32 psoriatic patients with no topical treatment in the intertriginous areas, from 13 psoriatic patients treated with topical steroids and from 19 patients with no psoriasis or other affections of the skinfolds. Untreated psoriatic patients were colonized by S. aureus significantly more often than the control group but infection seemed to be unlikely. Candida was not found in any of the groups. It is proposed that intertriginous psoriasis be treated with topical steroids alone and that the routine use of antimycotic and antibacterial combinations should be avoided. PMID- 12735641 TI - Detection of herpes virus genomes in skin lesions from patients with Behcet's disease and other related inflammatory diseases. AB - Although the aetiology of Behcet's disease is still poorly understood, viral infection has long been postulated as one of the factors. To investigate the relationship between herpes viruses and Behcet's disease, we used polymerase chain reaction to detect herpes simplex virus 1 (HSV-1) and 2 (HSV-2), and human herpes virus 6 (HHV-6) and 7 (HHV-7) DNA in samples of lesional tissues from patients with Behcet's disease and other related inflammatory disorders. Four cases were positive for HSV-1; 1 of 11 Behcet's disease cases, 2 of 3 Sweet's disease cases and 1 of 3 erythema nodosum cases. Two cases were positive for both HSV-1 and HSV-2; one Behcet's disease and one erythema nodosum. All cases were negative for HHV-6 and HHV-7. These findings indicate that there might be some relationship between Behcet's disease and the presence of HSV-1 and/or HSV-2 DNA and that HHV-6 and HHV-7 do not seem to be involved in the pathogenesis of Behcet's disease. However, HSV-1 and HSV-2 DNAs were also detected in non Behcet's disease lesions, suggesting that HSV-1/2 is not correlated to the direct pathogenesis of Behcet's disease. PMID- 12735643 TI - Lupus vulgaris in a skin graft. PMID- 12735642 TI - Human herpes virus 6 encephalitis in allopurinol-induced hypersensitivity syndrome. AB - Hypersensitivity syndrome is one of the most severe forms of drug eruption, and is characterized by a severe, potentially lethal, multiorgan involvement. Recently, reactivation of human herpesvirus 6 (HHV-6) has been suggested to be involved in this syndrome, although the exact role of HHV-6 remains elusive. In addition to exanthem subitum, neurological illnesses, such as infantile febrile illness without rash and encephalitis in immunocompromised patients have been attributed to HHV-6. A 51-year-old man developed a generalized erythematous eruption during treatment with allopurinol. Prednisolone improved his condition, but after the dose of prednisolone was reduced neurological abnormalities such as mental deterioration and positive meningeal signs developed. HHV-6 DNA in his blood by PCR analysis was positive. Furthermore, we detected HHV-6 DNA in the cerebrospinal fluid. The titers of anti-HHV-6-IgG increased during the course. His neurological symptoms gradually improved and no neurological sequelae were noted. Neurological abnormalities associated with hypersensitivity syndrome are very rare. However, the detection of HHV-6 DNA in the cerebrospinal fluid strongly indicates an involvement of reactivated HHV-6 in encephalitis. PMID- 12735644 TI - Erythema gyratum repens accompanied by essential thrombocythemia, followed by a blastic crisis. PMID- 12735645 TI - A de novo mutation in the keratin 9 gene in a family with epidermolytic palmoplantar keratoderma from northern Sweden. PMID- 12735646 TI - A novel insertion mutation in COL7A1 identified in Hallopeau-Siemens recessive dystrophic epidermolysis bullosa. PMID- 12735647 TI - A case of adult T-cell leukemia/lymphoma with an indolent clinical course has an unusual proviral DNA integration pattern. PMID- 12735648 TI - Severe mucocutaneous necrotizing vasculitis associated with the combination of chloroquine and proguanil. PMID- 12735649 TI - Lumbosacral hypertrichosis treated with the normal-mode ruby laser. PMID- 12735650 TI - Superinfection of eczema with multiple Acinetobacter species. PMID- 12735651 TI - Isolated plantar collagenoma. PMID- 12735653 TI - Long-term involution of unilateral porokeratotic eccrine ostial and dermal duct naevus. PMID- 12735652 TI - Severe generalized ulcerative lichen planus. PMID- 12735655 TI - Ulcerated necrobiosis lipoidica treated with cyclosporin A. PMID- 12735654 TI - Venous leg ulcers in a patient with Klinefelter's syndrome and increased activity of plasminogen activator inhibitor-1. PMID- 12735656 TI - Pyoderma gangrenosum associated with sarcoidosis. PMID- 12735657 TI - Divided naevus of the penis: a hypothesis on the embryological mechanism of its development. PMID- 12735658 TI - Adult multiple xanthogranulomas with spontaneous resolution. PMID- 12735660 TI - A single high dose of formoterol is as effective as the same dose administered in a cumulative manner in patients with acute exacerbation of COPD. AB - Several clinical trials have shown that the inhaled beta2-agonists with long acting properties, formoterol and salmeterol, may be effective in acute exacerbations of chronic obstructive pulmonary disease (COPD). However, there is a great deal of controversy regarding the timing and optimal dose of inhaled beta2-agonists in this pathologic condition. In this double-blind, randomised, crossover study, we have compared the bronchodilating effect and the safety of inhaled formoterol administered via Turbuhaler using either a cumulative dose regimen or the equivalent single dose in 16 patients with acute exacerbations of COPD. On the two consecutive days, the patients received, in a randomised order, each of the following active dose regimens: (A): 9 + 9 + 18 microg of formoterol via Turbuhaler (36 microg cumulative delivered dose) or (B): 36 + 0 + 0 microg of formoterol via Turbuhaler. The three doses on each treatment day were administered at 30-mm intervals, with measurements being made 5 and 30 min after each dose. Contemporaneously, we also measured oxygen saturation by pulse oximetry (SpO2) and pulse rate. Both the high dose and the cumulative one induced a significant bronchodilation expressed as change in FEV1. The difference between the two regimens was significant (P=0.0332) only 60 min after the first inhalation. The trend of FVC and IC was similar to that of FEV1. All treatment regimens were well tolerated and no adverse events were reported. Neither the administration ofthe high dose nor that of the cumulative one modified heart rate in a significant manner. Also they did not influence SpO2. This study indicates that a single high dose offormoterol is as effective as the same dose administered in a cumulative manner in patients with acute exacerbation of COPD. PMID- 12735659 TI - Bronchodilator effect of an inhaled combination therapy with salmeterol + fluticasone and formoterol + budesonide in patients with COPD. AB - In the present trial, we compared the broncholytic efficacy of the combination therapy with 50 microg salmeterol + 250 microg fluticasone and 12 microg formoterol + 400 microg budesonide, both in a single inhaler device, in 16 patients with moderate-to-severe COPD. The study was performed using a single blind crossover randomized study. Lung function, pulse oximetry (SpO2) and heart rate were monitored before and 15, 30, 60, 120, 180, 240, 300, 360, 480, 600, and 720 min after bronchodilator inhalation. Both combinations were effective in reducing airflow obstruction. FEV1 AUC(0-12 h) was 2.83 l (95% CI: 2.13-3.54) after salmeterol/fluticasone and 2.57 l (95% CI: 1.97-3.2) after formoterol/budesonide. Formoterol/budesonide elicited the mean maximum improvement in FEV1 above baseline after 120 min (0.29 l; 95% CI: 0.21-0.37) and salmeterol/fluticasone after 300 min (0.32 l; 95% CI: 0.23-0.41). At 720 min, the increase in FEV1 over baseline values was 0.10 l (95% CI: 0.07-0.12) after salmeterol/fluticasone and 0.10 l (95% CI: 0.07-0.13) after formoterol/budesonide. The mean peak increase in heart rate occurred 300 min after formoterol/budesonide (1.5 b/min; 95% CI--2.3 to 5.3) and 360 min after salmeterol/fluticasone (2.6 b/min; 95% CI--1.9 to 7.0). SpO2 did not change. All differences between salmeterol/fluticasone and formoterol/budesonide were not significant (P > 0.05) except those in FEV1 at 120 and 360 min. The results indicate that an inhaled combination therapy with a long-acting beta2-agonist and an inhaled corticosteroid appears to be effective in improving airway limitation after acute administration in patients suffering from COPD. PMID- 12735662 TI - The influence of co-morbidity on health-related quality of life in asthma and COPD patients. AB - This study examines the association between somatic co-morbidity and both general and disease-specific health-related quality of life (HRQoL) in patients with asthma and chronic obstructive pulmonary disease (COPD). A cross-sectional analysis was done among 161 COPD patients and 395 asthma patients, aged 40-75 years, recruited from general practice. In the total study population, 47% had no, 32% had one, and 21% had two or more somatic co-morbid conditions, with no significant differences between asthma and COPD patients. Co-morbidity appeared to be associated with poor disease-specific HRQoL in asthma [odds ratio (OR) = 2.08 (1.37-3.18)] and with poor general HRQoL in asthma [OR = 2.96 (1.93-4.53)] and COPD [1.81 (0.91-3.60)] patients. Poorest HRQoL was found in patients with more than one co-morbid condition. Cardiac disease and hypertension were associated with poor disease-specific HRQoL in asthma. Of all co-morbid conditions, musculoskeletal disorders were most strongly associated with poor general HRQoL. Cardiac disease was found to be associated with general and disease-specific HRQoL in asthma but not in COPD. In studies on patients with asthma or COPD aged 40-75 years, co-morbidity should be treated as a determinant of HRQoL. PMID- 12735661 TI - Fifty microg b.i.d. of inhaled fluticasone propionate (FP) are effective in stable asthmatics previously treated with a higher dose of FP. AB - Twenty-seven subjects with moderate asthma at the time of diagnosis, well controlled under regular fluticasone propionate (FP) (250 microg b.i.d.) for 6 months at least, were randomized to receive in double-blind fashion: FP 125 microg b.i.d. (Group 1) or FP 50 microg b.i.d. (Group 2) or placebo (Group 3) for 3 months or until symptom recurrence. Daily symptom score and peak expiratory flow were monitored. At the beginning and at the end of the study subjects underwent methacholine challenge and sputum induction. Recurrence of symptoms occurred shortly after randomization in all subjects receiving placebo. None from Group 1 or 2 experienced symptom recurrence during the study. No significant difference in clinical and functional data, and in sputum eosinophil percentages was observed between the beginning and the end of the study in both Groups 1 and 2. Subjects from Group 3 showed a significant increase of sputum eosinophils (P<0.05) and a significant decrease in provocative dose of methacholine (P<0.05) when asthma symptoms recurred. Therefore, very low doses of FP (50 microg b.i.d.) are effective in maintaining for 3 months a good control of the disease in asthmatics already stable under high-dose fluticasone, considering both clinical and functional outcomes and markers of airway inflammation. PMID- 12735663 TI - A comparison of the end-tidal CO2 measured by portable capnometer and the arterial PCO2 in spontaneously breathing patients. AB - An end-tidal CO2 (ETCO2) monitor (capnometer) is used most often as a noninvasive substitute for PaCO2 in anesthesia, anesthetic recovery and intensive care. However, the utility and accuracy of the portable capnometer in spontaneously breathing patients with or without chronic pulmonary diseases has received little recognition. To determine the utility of the portable capnometer in general wards and in in-home care, we examined the correlation between ETCO2 measured by a portable capnometer and simultaneous PaCO2 measured in 41 spontaneously breathing patients. TV-ETCO2 (ETCO2 measured by tidal volume maneuver) was lower than PaCO2 by an average of 9.0 mmHg and VC-ETCO2 (ETCO2 measured by vital capacity maneuver) was lower than PaCO2 by an average of 0.5 mmHg. The mean difference between PaCO2 and VC-ETCO2 was not statistically significant. Regression analysis showed a close correlation between VC-ETCO2 and PaCO2 (r = 0.91, P < 0.0001). Thus,VC-ETCO2 was highly correlated with PaCO2. Furthermore, a close correlation between VC-ETCO2 and PaCO2 was also observed in patients with compromised pulmonary function (r = 0.88, P < 0.0001 in patients with below 70% of FEV(1.0)%; r = 0.89, P < 0.0001 in patients with below 80% of %VC). Our studies show that VC ETCO2 measured by the portable capnometer gives a reliable pointestimate of PaCO2, and can be useful to evaluate the respiratory condition of spontaneously breathing patients in general wards and in in-home care. PMID- 12735664 TI - Impairments and prognostic factors for survival in patients with idiopathic pulmonary fibrosis. AB - The aim of the present study was to evaluate exercise limiting factors using cardiopulmonary exercise testing (CPET) in patients with idiopathic pulmonary fibrosis (IPF), and to investigate whether these parameters are related to survival after CPET. We evaluated 41 patients with IPF (mean 68.2 years, 27 male) who performed CPET. The exercise capacity in patients with IPF was limited more strongly by gas exchange and/or ventilatory impairments, compared with cardiac impairment. Using univariate analysis, the severity of exercise-induced hypoxemia (EIH) evaluated by deltaPaO2/deltaVO2 (PaO2-slope), oxygen uptake at maximum exercise, oxygen pulse at maximum exercise, ventilatory equivalent for carbon dioxide at maximum exercise and age were significantly related to the survival rate. Interestingly, the PaO2-slope was most closely correlated with the survival rate using multiple analysis with a stepwise evaluation. Nevertheless, PaO2 at rest and at maximum exercise were not factors influencing survival. In patients with IPF, CPET can simultaneously evaluate the ability of both the cardiovascular and respiratory systems, and should be available so that parameters can be derived to make the necessary prognostic estimations, with the most useful parameter being the degree of EIH as represented by the PaO2-slope. PMID- 12735665 TI - Gender differences in asthma prevalence may depend on how asthma is defined. AB - BACKGROUND: Asthma may be defined either as wheeze within the previous 12 months (current wheeze) doctor-diagnosed asthma (DDA), or current wheeze plus confirmed airway hyperresponsiveness (AHR). AIMS: We wanted to estimate asthma prevalence in randomly selected adolescents based on different criteria for asthma diagnosis, study gender differences in reported asthma-like symptoms vs DDA, and relate our findings to measurements of AHR, levels of exhaled nitric oxide (ENO) and total IgE. METHODS: As part of the health survey of North-Trondelag (HUNT), 8571 adolescents aged 13-19 years were investigated with an interview on allergic and respiratory symptoms (phase I study). Of these, 401 subjects who reported wheeze within the previous 12 months (current wheeze) and 213 non-symptomatic controls were randomly selected and investigated with allergy screening, methacholine bronchoprovocation test and measurements of ENO (phase II study). RESULTS: In the phase I study prevalence of current wheeze was 26% (30% in girls and 23% in boys, P < 0.01). Prevalence of DDA was 10.8% (10.5% in girls and 11% in boys). Among subjects with current wheeze, the likelihood of having DDA was reduced in girls compared to boys, odds ratio (95% CI) 0.82 (0.68-0.98) which was partly explained by a longer history of wheeze among boys. In the phase II study, although more girls than boys with current wheeze had AHR (62% versus 50%, P < 0.02) more boys than girls reported DDA (44% vs. 32%, P < 0.02). Of the objective parameters, increased levels of ENO most strongly increased the risk of having DDA. CONCLUSIONS: When asthma is defined as DDA, there is a risk of underestimating the prevalence of asthma, especially among girls. PMID- 12735666 TI - Low-dose methotrexate for advanced pulmonary disease in patients with cystic fibrosis. AB - Inflammation is a hallmark in the pathogenesis of pulmonary destruction in cystic fibrosis (CF). There is no proven effective systemic anti-inflammatory treatment for CF patients with advanced pulmonary disease. Methotrexate (MTX) is known as an effective anti-inflammatory treatment in asthma and in juvenile rheumatoid arthritis. The question was: Is an improvement in pulmonary function achievable with low-dose MTX in patients with cystic fibrosis and advanced pulmonary disease.? METHODS: We treated five CF patients with advanced pulmonary disease, who deteriorated in spite of intensive conventional therapy on an individual basis with low-dose MTX. FEV1% and immunoglobulin G (IgG) serum levels were followed from the year before to the year after starting with MTX. RESULTS: In the year before starting with MTX, FEV1% decreased (median: 10% FEV1; range 9-15% FEV1; P<0.005) after starting with MTX, FEV1% increased (median: 9% FEV1; range: 2-15% FEV1; P<0.05). IgG changed (median: -2 g/l; range: 0.2 to -7.3 g/l) in the first year with MTX. CONCLUSION: These preliminary data suggest a beneficial effect of MTX even in advanced pulmonary disease in CF patients and supports the need for a controlled prospective study. PMID- 12735667 TI - Breathing retraining for asthma. AB - Breathing retraining is used increasingly throughout the world by many patients with asthma in addition to their usual medical care. We undertook a systematic review of the literature in order to determine the effectiveness of breathing retraining in the management of asthma. Six randomised-controlled trials were identified that involved breathing retraining in asthma. Due to the variation in reported trial outcomes, limited reporting of study data and small number of included trials it was not possible to draw any firm conclusions as to its effectiveness. However, outcomes that were reported from individual trials do show that breathing retraining may have a role in the treatment and management of asthma. Further large-scale trials using breathing retraining techniques in asthma are required to address this important issue. PMID- 12735669 TI - Interleukin-18 expression in BAL cells of sarcoidosis patients is decreased in vivo but protein secretion is not impaired in vitro. AB - Recently, increased expression of interleukin-18 (IL-18) has been shown in sarcoid airway epithelium. However, IL-18 expression has not been investigated extensively in bronchoalveolar lavage (BAL) cells in sarcoidosis yet. Expression of IL-18 and tumour necrosis factor-alpha (TNFalpha) mRNA in cells of the BAL of 23 patients with sarcoidosis and nine healthy volunteers was determined using semiquantitative RT-PCR. IL-18 protein in BAL cells was investigated by immunocytochemistry (ICC). IL-18 protein levels in BAL cell culture supernatants from patients and controls with and without LPS stimulation were measured by enzyme-linked immuno-sorbent assay. BAL cells from patients were stimulated with either IL-10 or IL-13 and IL-18 protein levels were determined. IL-18 mRNA expression was significantly decreased in BAL cells of patients compared to control subjects (1.62 +/- 0.27 vs. 4.29 +/- 0.77, P < 0.05). TNFalpha mRNA expression was significantly increased in BAL cells of patients in comparison to control subjects (0.63 +/- 0.09 vs. 0.11 +/- 0.08, P < 0.05). ICC showed less positive alveolar macrophages in sarcoidosis patients than in control subjects (26 vs. 53%). IL-18 protein levels did not differ significantly between both groups. Stimulation with IL-10 significantly reduced IL-18 protein concentration in sarcoidosis patients. Our results suggest that BAL cells may not be the main source of IL-18 production in sarcoidosis in vivo. Since IL-18 production of BAL cells was not impaired in culture antiinflammatory cytokines might contribute to decreased IL-18 expression in vivo. PMID- 12735668 TI - Measuring costs and consequences in economic evaluation in asthma. AB - Formal economic evaluation is playing an increasingly important role in health care decision-making. This is shown by the requirement to present economic data to support applications for public reimbursement for new pharmaceuticals in Australia and the provinces of Canada, and by the appraisal process initiated by the National Institute for Clinical Excellence in the U.K. This growing role of economic analysis applies as much to the field of asthma as anywhere. This paper provides a detailed review of applied economic studies in asthma. The review is used to explore a range of methodological issues in the field including the choice of perspective and maximand, whether to use disease-specific or generic measures of outcome and whether decision-makers should receive disaggregated cost and consequence data or results that focus on an incremental cost-effectiveness ratio. It is concluded that, given the heterogeneity in decision-makers' objectives and constraints, economic studies should be planned and executed in such a way as to maximize flexibility in how results are presented. PMID- 12735670 TI - Potential utility of BCG test in excluding tuberculosis in adults. AB - In developing countries, many sputum-negative suspects of pulmonary tuberculosis (PTB) are given antituberculous medication just because tuberculosis (TB) could not be excluded in them. Many studies have shown that BCG test is nearly 100% sensitive, except in those with severe malnutrition, steroid therapy and other causes of immunosuppression. The aim of the present study was to find out the proportion of PTB suspects in whom BCG test may be useful in excluding diagnosis of TB. The study was carried out on adult subjects attending District Tuberculosis Clinic (DTC), Pali, Rajasthan, India. Out of 104 subjects studied, all 18 sputum-positive subjects showed a positive BCG test indicating 100% sensitivity of the test. The result of BCG test was negative in 16.28% of 86 sputum-negative suspects of TB. The BCG test did not show any adverse reactions among these cases except for adenitis of axillary or cervical lymph nodes in 6.7% subjects. The study raises the possibility that BCG test may be useful in excluding diagnosis of tuberculosis in a proportion of adult sputum negative suspects of PTB in developing countries like India. More studies are needed in this direction. PMID- 12735671 TI - Effects of oral montelukast on airway function in acute asthma. AB - Montelukast, a specific cysteinyl leukotriene receptor antagonist, has been shown to improve pulmonary function within 1 h of ingestion. This study was undertaken to compare the effects on peak expiratory flow rate (PEFR) of oral montelukast added to intravenous steroid, intravenous steroid alone and placebo during the 24 h period following administration. Seventy asthmatic patients (FEV1 40-80% predicted and > or = 15% improvement after inhaled beta agonist) were enrolled in a single blind study to receive oral montelukast (10 mg) plus intravenous prednisolone (1 mg/kg), intravenous prednisolone (1 mg/kg) or placebo in a randomised fashion. The patients received one ofthe above three groups of medication before any other treatments. This was immediately followed by the aerosol treatments of 100 mcg of terbutaline sulphate divided into three doses during 1 h as described in the consensus statement. Thereafter, patients were observed for 24 h to document the effects on PEFR, Borg dyspnoea score and need for rescue medication. The primary end point was percentage change at different time points. Secondary end points were Borg dyspnoea score and use of rescue medication. Compared with placebo, montelukast added to the prednisolone group and the prednisolone alone group had significant percentage change from baseline in PEFR in the entire 24 h period (P<0.05). The difference in PEFR between montelukast plus prednisolone group and prednisolone group favoured the montelukast plus prednisolone group but did not reach statistical significance. Furthermore, montelukast plus prednisolone group required less inhaled short acting beta agonistthan other two groups. The results of this study indicate that adding montelukast to steroid in acute asthma may have some additive improvement in lung functions. PMID- 12735672 TI - Overnight home oximetry: can it identify patients with obstructive sleep apnea hypopnea who have minimal daytime sleepiness? AB - BACKGROUND: Overnight home oximetry is being widely used as a case selection technique for patients with suspected obstructive sleep apnea-hypopnea (OSAH). In the absence of excessive daytime sleepiness (EDS), patients with snoring and/or witnessed apnea are considered to have low probability of OSAH. METHODS: Patients suspected to have OSAH, who denied EDS, and had a normal overnight home oximetry were studied by polysomnography for presence of significant OSAH (apnea/hypopnea index (AHI) > 15/h). RESULTS: Twelve (40%) of the 30 patients studied had significant OSAH. All had a 2% oxygen desaturation index of less than 10/h. The sensitivity of oximetry increased at lower desaturation indices butthis was associated with decreased specificity. Review of oximetry waveform pattern, by experienced physicians, did not improve the diagnostic accuracy. Combining oximetry with a clinical prediction rule would have reduced the need for polysomnography by 30%. CONCLUSION: Many patients, who present with snoring and/or witnessed apnea and are referred to a sleep disorder clinic for suspected OSAH, may have significant OSAH even if they deny EDS. Overnight home oximetry did not help discriminate between those patients with or without OSAH. PMID- 12735673 TI - Relationship of asthma to irritant gas exposures in pulp and paper mills. AB - The potential of chronic or acute irritant gas exposures to cause asthma or a variant condition, reactive airways dysfunction syndrome (RADS) was investigated by observing asthma incidence in a large working population, using person-years at risk (P-YR) to compute relative rates (RR). Health data came from employee examinations at 62 pulp and paper plants. The 39122 workers who denied asthma beginning before the observation period included: 19326 denying irritant exposures, with no gassing exposures; 19349 with self-reported irritant exposures, and no gassing; and 447 with documented gassings. Asthma was defined as self-reported asthma beginning after the start of observation. P-YR accrued from September 29, 1986, for the nonexposed and exposed workers, and from date of first gassing for gassed workers, and ended with disease onset in any who developed asthma. RR of asthma with 95% confidence intervals (CI95) were calculated for the exposed and gassed groups, relative to the nonexposed. Exposed (nongassed) workers had an elevated asthma rate, RR=1.48, CI95=1.17-1.86, after adjustment for effects of gender and number of examinations. The rate in gassed workers was not significantly elevated: RR=1.95, CI95=0.75-5.08. Of the five asthma cases occurring after gassings, none conformed to diagnostic criteria for RADS. Chronic exposures were associated with increased rate of asthma onset, which must be interpreted with caution because self-reported data defined both exposure category and disease. Documented gassings were not associated with significantly increased rate, and none of 447 gassed persons developed RADS. PMID- 12735674 TI - Scintigraphic diagnosis of a right to left shunt in end-stage lung disease. AB - PURPOSE: The presence of a right to left shunt influences the surgical approach to lung transplantation in patients with end-stage pulmonary disease. The purposes of this study included comparing contemporaneous lung scintigraphy with cardiac catheterization in the detection of intracardiac shunts in patients with end-stage lung disease and the point prevalence of right to left shunting was determined in patients with several different types of end-stage lung disease. METHODS: Hundred and twenty six patients with end-stage lung disease who were candidates for lung transplantation underwent perfusion images of the lungs with Tc-99m-labeled macro-aggregated albumin (MAA). Planar scans of the brain and the kidneys were performed contemporaneously. Statistical analyses included correlation ofthe clinical, laboratory and scintigraphic variables. Group means were compared with the students t-test (two-tailed P-value). RESULTS: There were 21 patients with primary pulmonary hypertension (PPH), 72 with emphysematous lung disease (COPD), 22 with pulmonary fibrotic disease (PF) and 11 with congenital heart disease (CHD) leading to pulmonary hypertension. Only 13 patients (10.3%) were found to have a right to left shunt. Of these, 4 had PPH, 2 had PF, and 7 had CHD. No shunts were found in patients with emphysema. All the positive studies had abnormally increased activity in both the brain and the kidneys. However, there were 25 cases with renal activity and none of these patients had brain activity or clinical evidence of a shunt. Increased pulmonary artery pressure was associated with scintigraphic presence of a shunt. There were no cases of a right to left shunt with a mean pulmonary artery pressure less than 50 mm Hg. In the subset of patients with a pulmonary pressure greater than 50 mm Hg, approximately 40% of the patients had a right to a left shunt. There were no measurable differences in the spirometry results, right ventricular ejection fraction (RVEF) or left ventricular ejection fraction (LVEF) in the subgroup of patients with PPH and right to left shunt in comparison with patients with PPH but without a right to left shunt. CONCLUSIONS: The findings indicate that images of the brain, but not the kidneys, are an effective way to diagnose extrapulmonary right to left shunts in patients with end-stage pulmonary disease. The problem of a right to left shunt is uncommon in patients with emphysematous lung disease and relatively common in patients with primary pulmonary hypertension. PMID- 12735675 TI - Addition of salmeterol to fluticasone propionate treatment in moderate-to-severe asthma. AB - This study was designed to determine whether the benefit of adding salmeterol was superior to doubling the dose of fluticasone propionate (FP) over 6 months, compared to a control group who remained on a lower dose of FP. The multi-centre, double-blind, parallel group study involved 496 symptomatic asthmatic patients with a history of exacerbations on 500-800 micrograms (microg) inhaled corticosteroids (ICS) twice daily (b.d.) in a broadly representative group of 100 hospitals and general practices in six countries. Two doses of FP--250 microg b.d. (FP250) or 500 microg b.d. (FP500)--were compared with the lower dose of FP plus a long-acting beta2-agonist, salmeterol 50 microg b.d. (SM/FP250). Patients symptomatic on the run-in dose of FP250 alone formed the control group in the treatment period. Over 6 months, SM/FP250 significantly improved mean morning peak expiratory flow rates (amPEF) by 42.1 l/min, more than twice the improvement achieved with either dose of FP alone. SM/FP250 also resulted in more symptom free days and nights (P < 0.002) and days and nights with no relief medication (P < 0.001). The number of severe exacerbations was low: 3, 6 and 8% in the SM/FP250, low- and high-dose FP groups, respectively. This study confirms that adding salmeterol to low-dose inhaled FP offers greater improvements than either maintaining or doubling the dose of FP. Significant benefit was gained from adding salmeterol in a group of patients who appeared to have been at the top of their steroid dose-response curve receiving FP250. There was no evidence of tolerance and a low incidence of exacerbations in all treatment groups. PMID- 12735676 TI - Expression of matrix metalloproteinases MMP-9 within the airways in asthma. AB - The matrix metalloproteinase (MMP) enzymes MMP-9, have relevance to chronic structural airway changes in asthma, which can be generated by structural and inflammatory cells, and have the ability to degrade proteoglycans and thus potentially enhance airway fibrosis and smooth muscle proliferation through their ability to release and activate latent, matrix-bound growth factors. Immunostaining for MMP-9 was undertaken in acetone-fixed and glycolmethacrylate embedded endobronchial biopsy specimens obtained by fibreoptic bronchoscopy under local anaesthesia. The findings from 30 asthmatic subjects were compared with those from 18 chronic obstructive pulmonary disease (COPD) subjects and 10 healthy controls. Meanwhile, pulmonary function test and airway responsiveness were performed. Immunoreactivity for MMP-9 was assessed by an image analysis system. The biopsy specimens from asthmatic subjects contained significantly more eosinophils (P < 0.001) than those from COPD subjects, and healthy control did not contain eosinophils. MMP-9 immunoreactivity could be identified in endobronchial biopsy specimens from all the asthmatic subjects and 40% ofthe COPD subjects, but could not be identified in healthy controls. Gelatinase B (MMP-9) immunoreactivity was located in bronchial epithelium and extracellular matrix in submucosa, prominent in denuded epithelium. The immunohistochemical score for MMP 9 was significantly correlated with eosinophilic number in bronchial mucosa. FEV1% predicted FEV1/FVC (%) (r = 0.52, 0.41, 0.37, respectively P < 0.01 did not correlate with PD20 FEV1 from asthmatic subjects. MMP-9 is expressed by bronchial epithelium and may be a important factor for eosinophil infiltraed into airway from asthma subjects. PMID- 12735678 TI - The use of inhaled corticosteroids in the United Kingdom and the Netherlands. AB - This study examined the utilisation patterns of inhaled corticosteroids in England/Wales and the Netherlands. Computerised medical records from the GPRD (U.K.) and PHARMO (the Netherlands) databases were used. It included 284733 English/Welsh and 27761 Dutch adult patients who were prescribed inhaled corticosteroids during the 10-year study period. Our results showed that, in both study populations, overall use of inhaled corticosteroids increased over the period studied, with its prevalence rising steeply with age and declining in extreme old age. Decreased use of bronchodilators and oral corticosteroids in the early treatment of asthma was noted in our findings. In addition, a trend towards the decreasing use of oral corticosteroids concomitant with inhaled corticosteroid therapy was also observed for both groups. Our study found that only 42.1% of the GPRD and 31.1% of the PHARMO patients received a repeat prescription within the expected duration ofthe preceding inhaled corticosteroid prescription. In conclusion, our study found many similarities in the prescribing and use of inhaled corticosteroids between the two study populations. The observation of irregular use of inhaled corticosteroid among a substantial number of patients highlights a need for further study into the reasons for irregular use and its consequences on the effectiveness of treatment. PMID- 12735677 TI - Reproducibility of forced inspiratory and expiratory volumes after bronchodilation in patients with COPD or asthma. AB - The aim of the present study was to assess the reproducibility of changes in forced inspiratory volumes after bronchodilator inhalation. Thirteen patients with chronic obstructive pulmonary disease (COPD) (FEV1, 32-75%pred) and 10 patients with asthma (FEV1, 43-75%pred) inhaled either 200 microg fenoterol or 200 microg oxitropium bromide or placebo, each of them on three occasions, on nine different days in a randomised, cross-over, double-blind fashion. Forced expiratory (FEV1) and inspiratory (FIV1) volumes were measured before and 30 min after inhalation. In patients with COPD, the increase in FEV1 (coefficient of variation) was 221 ml (43%) after fenoterol and 235 ml (33%) after oxitropium; changes in FIV1 were 301 ml (45%) and 360 ml (29%). In patients with asthma, FEV1 improved by 618 ml (26%) and 482 ml (25%), FIV1 by 553 ml (41%) and 475 ml (23%). In less severe COPD or asthma, the reduction in dyspnoea was associated with the improvements in both FIV1 and FEV1, but in severe COPD with the improvement in FIV1 only. The data demonstrate that, at least in terms of relative changes, the reproducibility of bronchodilator responses in terms of FIV1 is similar to that of FEV1 and they underline the assertion of FIV1 being a sensible parameter particularly in severe COPD. PMID- 12735679 TI - Apparent primary pleural melanoma: case report and literature review. PMID- 12735680 TI - Synthesis and evaluation of cytotoxicity of stilbene analogues. AB - Resveratrol analogs were newly synthesized and evaluated for cytotoxicity in cultured human lung and colon cancer cells. 3,5,4-Trimethoxy-trans-stilbene and 3,5,2',4'-tetramethoxy-trans-stilbene were found to be more potent rather than resveratrol. 3,4,5-Trimethoxy-4'-bromo-cis-stilbene was the most active among the test compounds. PMID- 12735681 TI - Synthesis and in vitro properties of prednisolone 21-sulfate sodium as a colon specific prodrug of prednisolone. AB - Colon-specific delivery of glucocorticoids is highly desirable for the efficient treatment of inflammatory bowel disease. We synthesized prednisolone 21-sulfate sodium (PDS) as a colon-specific prodrug of prednisolone (PD) and investigated its properties using rats as test animals. We expected that introduction of sulfate ester as a sodium salt might increase the hydrophilicity and restrict the absorption in the GI tract. If PDS is stable and nonabsorbable in the upper intestine, it will be delivered to the colon as an intact form, where it hydrolyze by the sulfatase to release PD. Compared with PD, the solubility of PDS increased and the apparent partition coefficient decreased greatly. PDS was stable on incubation with pH 1.2 and 6.8 buffer solutions and with the contents of the stomach and small intestine. On incubation with the cecal contents, PDS decreased to 9.6% of the dose in 10 h producing PD. The amount of PD increased to give a maximum 54% of the dose and decreased. As a control, when PD was incubated with the cecal contents, it decreased to 29% of the dose in 8 h, which implied that reduction of PD proceeded under such conditions. These results suggested that hydrolysis of PDS took place to produce and accumulate PD, which decreased by reduction as the incubation period extended. Our results suggested that PDS can be a promising colon-specific prodrug of PD, and sulfate ester group might serve as a potential colon-specific promoiety, especially for the drugs which are resistant to reduction in the colon. PMID- 12735682 TI - Synthesis and properties of 5-aminosalicyl-taurine as a colon-specific prodrug of 5-aminosalicylic acid. AB - 5-Aminosalicylic acid (5-ASA) is an active ingredient of therapeutic agents used for Crohn's disease and ulcerative colitis. Because it is absorbed rapidly and extensively in the upper intestine, delivery of the agent specifically to the colon is necessary. We selected taurine as a colon-specific promoiety and designed 5-aminosalicyltaurine (5-ASA-Tau) as a new colon-specific prodrug of 5 aminosalicylic acid (5-ASA). It was expected that introduction of taurine would restrict the absorption of the prodrug and show additive effect to the anti inflammatory action of 5-ASA after hydrolysis. 5-ASA-Tau was prepared in good yield by a simple synthetic route. The apparent partition coefficient of 5-ASA Tau in 1-octanol/pH 6.8 phosphate buffer or CHCl3/pH 6.8 phosphate buffer was 0.10 or 0.18, respectively, at 37 degrees C. To determine the chemical and biochemical stability in the upper intestinal environment, 5-ASA-Tau was incubated in pH 1.2 and 6.8 buffer solutions, and with the homogenates of tissue and contents of stomach or small intestine of rats at 37 degrees C. 5-ASA was not detected from any of the incubation medium with no change in the concentration of 5-ASA-Tau. On incubation of 5-ASA-Tau with the cecal and colonic contents of rats, the fraction of the dose released as 5-ASA was 45% and 20%, respectively, in 8 h. Considering low partition coefficient and stability in the upper intestine, 5-ASA-Tau might be nonabsorbable and stable in the upper intestine. After oral administration, it would be delivered to the colon in intact form and release 5-ASA and taurine. These results suggested 5-ASA-Tau as a promising colon specific prodrug of 5-ASA. PMID- 12735683 TI - Synthesis and cytotoxicity of new 3-alkyl-1-(1-methyl-2-phenylethyl)ureas related to ceramide. AB - A series of new 3-alkyl-1-(1-methyl-2-phenylethyl)ureas related to ceramide was synthesized and evaluated for their in vitro cytotoxic activity against five human tumor cell lines. The urea analogue (2b) of B13 showed comparable or slightly more potent cytotoxic activity as compared to B13, indicating that urea does appear to serve as a bioisostere of amide. PMID- 12735684 TI - A new acylglycosyl sterol from Quisqualis Fructus. AB - A new acylglycosyl sterol (4) was isolated from the MeOH extract of Quisqualis Fructus together with four known compounds. On the basis of spectroscopic data, their structures were elucidated as clerosterol (1), betulinic acid (2), methylursolate (3), 3-O-[6'-O-(8Z-octadecenoyl)-beta-D-glucopyranosyl] clerosterol (4) and alpha-xylofuranosyluracil (5). PMID- 12735685 TI - Antioxidant principles of Nelumbo nucifera stamens. AB - In our ongoing study to identity antioxidants from natural sources, the antioxidant activity of Nelumbo nucifera stamens was evaluated for their potential to scavenge stable 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radicals, inhibit total reactive oxygen species (ROS) generation, in kidney homogenatas using 2',7'-dichlorodihydrofluorescein diacetate (DCHF-DA), and scavenge authentic peroxynitrites (ONOO-). A methanol (MeOH) extract of the stamens of N. nucifera showed strong antioxidant activity in the ONOO- system, and marginal activity in the DPPH and total ROS systems, so were therefore fractionated with several organic solvents, such as dichloromethane (CH2Cl2), ethyl acetate (EtOAc) and n-butanol (n-BuOH). The EtOAc soluble fraction, which exhibited strong antioxidant activity in all the model systems tested, was further purified by repeated silica gel and Sephadex LH-20 column chromatographies. Seven known flavonoids [kaempferol (1), kaempferol 3-O-beta-D-glucuronopyranosyl methylester (2), kaempferol 3-O-beta-D-glucopyranoside (3), kaempferol 3-O-beta-D galactopyranoside (4), myricetin 3',5'-dimethylether 3-O-beta-D-glucopyranoside (5), kaempferol 3-O-alpha-L-rhamnopyranosyl-(1-->6)-beta-D-glucopyranoside (6) and kaempferol 3-O-beta-D-glucuronopyranoside (7)], along with beta-sitosterol glucopyranoside (8), were isolated. Compound 1 possessed good activities in all the model systems tested. Compounds 2 and 7 showed scavenging activities in the DPPH and ONOO- tests, while compounds 3 and 4 were only active in the ONOO- test. Conversely, compound 8 showed no activities in any of the model systems tested. PMID- 12735686 TI - Antioxidative properties of brown algae polyphenolics and their perspectives as chemopreventive agents against vascular risk factors. AB - Several polyphenolic compounds and complex mixtures were isolated from brown algae species. The 1,1-diphenyl-2-picryhydarzyl (DPPH) radical scavenging activity and ferric reducing antioxidant power (FRAP) of these compounds were evaluated to determine their physiological usefulness as antioxidants for vascular protection. The antioxidative protection of low-density lipoprotein (LDL) was also evaluated and compared with that of catechin, because the generation of oxidized LDL is one of the most active and specific risk factors contributing to atherogenesis. Oral administration to rats of a commercially available sample (VNP) containing 30% of these polyphenolic compounds and 70% dietary fiber revealed that the serum reducing capacity measured in terms of FRAP value was significantly elevated 30 min after the treatment, but declined rather quickly thereafter, indicating the good oral absorption of the compounds and their fast binding to the lumenal surface of the blood vessels. An eight-week, human, clinical trial (n=31) of VNP showed significant improvement in erectile function measured by IIEF (international index of erectile function) score. These results collectively demonstrated the usefulness of these polyphenolic compounds as fundamental chemopreventive agents against vascular risk factors originating from oxidative stress. PMID- 12735687 TI - The polysaccharide fraction AIP1 from Artemisia iwayomogi suppresses apoptotic death of the mouse spleen cells in culture. AB - A polysaccharide fraction, AIP1, purified from Artemisia iwayomogi was shown to have immunomodulating and anti-tumor activities in mice. In order to determine how the AIP1 fraction exhibits the immunomodulating activity, the effect of the fraction on the apoptosis of mouse spleen cells was investigated. Treatment of the mouse spleen cells with the AIP1 fraction resulted in the suppression of apoptotic death and an extension of cell survival in culture, indicating that the fraction might modulate the death of spleen cells. Treatment of the mice with the AIP1 fraction in vivo also resulted in less apoptosis of the spleen cells, which indicates the physiological relevance of the anti-apoptosis effect of the fraction in vitro. A mouse gene array was used to determine the profile of the gene expression change showing a pattern of up- and down-regulated genes by the AIP1 treatment. This study provides preliminary information regarding the immunomodulatory mechanism of the AIP1 fraction. PMID- 12735688 TI - Inhibitors of inducible nitric oxide synthase expression from Artemisia iwayomogi. AB - Nitric oxide (NO) is an important bioactive agent that mediates a wide variety of physiological and pathophysiological events. NO overproduction by inducible nitric oxide synthase (iNOS) results in severe hypotension and inflammation. This investigation is part of a study to discover new iNOS inhibitors from medicinal plants using a macrophage cell culture system. Two sesquiterpenes (1 and 2) were isolated from Artemisia iwayomogi (Compositae) and were found to inhibit NO synthesis (IC50 3.64 microg/mL and 2.81 microg/mL, respectively) in lipopolysaccharide (LPS)-activated RAW 264.7 cells. Their structures were identified as 3-O-methyl-isosecotanapartholide (1) and iso-secotanapartholide (2). Compounds 1 and 2 inhibited the LPS-induced expression of the iNOS enzyme in the RAW 264.7 cells. The inhibition of NO production via the down regulation of iNOS expression may substantially modulate the inflammatory responses. PMID- 12735690 TI - Prevention of macrophage-related inflammatory diseases by allergina. AB - The oriental herbal combination allergina has been shown to inhibit allergic inflammation. In the present study, we demonstrate that the oral administration of allergina markedly inhibits the progression of inflammatory diseases, such as graft-versus-host diseases (in the allogeneic bone marrow transplantation and the parent-into-F1 transplantation models), collagen-induced arthritis and sheep red blood cell-induced delayed type hypersensitivity. The immunosuppressive activity of allergina in vivo appears to be associated, at least in part, with the inhibition of tumor necrosis factor-alpha production. In conclusion, our results suggest that allergina could be useful as a immunosuppressive agent for the treatment of macrophage-related inflammatory disease. PMID- 12735689 TI - Anti-inflammatory mode of isoflavone glycoside sophoricoside by inhibition of interleukin-6 and cyclooxygenase-2 in inflammatory response. AB - Soy, high dietary intake for the oriental population, is a main source of isoflavonoids. Sophoricoside (SOP) an isoflavone glycoside was isolated from immature fruits of Sophora japonica (Leguminosae family) and its inhibitory effect on chemical mediators involved in inflammatory response was investigated in this study. SOP inhibited the interleukin (IL)-6 bioactivity with an IC50 value of 6.1 microM whereas it had no effects on IL-1beta and TNF-alpha bioactivities. SOP was identified as a selective inhibitor of cyclooxygenase (COX)-2 activity with an IC50 value of 4.4 microM, but did not show inhibitory effect on the synthesis of COX-2. However, SOP had no effect on the production of reactive oxygen species including superoxide anions and nitric oxide. These results revealed that in vitro anti-inflammatory action of SOP is significantly different from that of genistein known as a phytoestrogen of soy products. This experimental study has documented an importance of dietary soy isoflavonoids as multifunctional agents beneficial to human health, and will help to clarify protective mechanisms of SOP against inflammatory conditions. PMID- 12735691 TI - Lidocaine intoxication: two fatal cases. AB - We present two fatal cases, a 41-year-old male (case 1) and his 8-year-old daughter (case 2), resulting from acute lidocaine poisoning. Lidocaine was quantified by gas chromatography (GC) analysis using nitrogen-phosphorus detection. The lidocaine concentrations of cases 1 and 2 were: liver, 27.7 microg/g and 24.9 microg/g; spleen, 70.1 microg/g and 29.9 microg/g; and gastric contents, 23.6 microg/g and 42.8 microg/g, respectively. PMID- 12735692 TI - Chronic exposure of nicotine modulates the expressions of the cerebellar glial glutamate transporters in rats. AB - Rats were given nicotine (25 ppm) in their drinking water at the start of their mating period in order to study the expressions of glutamate transporter subtypes in cerebellar astrocytes following the chronic exposure of nicotine after mating. After the offspring were delivered, each group was divided into two subgroups. One group, the control group, was given distilled water only and the other group, the experimental group, was given distilled water containing nicotine. The cerebellar astrocytes were prepared from 7 day-old pups at each group. Ten days after the cells were cultured, the expression of the glutamate transporter subtypes (GLAST and GLT-1) was determined using immunochemistry and immunoblotting. During the continuous treatments, the developmental expression patterns of the GLAST and GLT-1 in the cerebellum were also determined from 2, 4 and 8 week-old rats. The expression levels of GLAST in cultured astrocytes of both the pre- or post-natally exposed groups were higher than those of the control group. However, these expression levels of the continuously exposed group were lower than those of the control group. Compared to those of the control group, the GLT-1 expression levels of all the nicotine-treated groups were higher, particularly in the continuously treated group. According to the results from the immochemistry procedure, the cerebellar GLAST and GLT-1 expression levels of all nicotine-treated groups were lower than those of the control group at each age. However, the immunoblotting procedure showed that the cerebellar GLT 1 expression levels of all the nicotine-treated groups were higher than those of the control group, except for the rats that were continuously exposed for 8 weeks using immunoblotting. These results suggest that the expression of the glial GLAST and GLT-1 are altered differently depending on the initial exposure time and the particular period of nicotine exposure. In addition, nicotine exposure during gestation has persistent effects on glial cells. PMID- 12735694 TI - Kinetic analysis about the bidirectional transport of 1-anilino-8-naphthalene sulfonate (ANS) by isolated rat hepatocytes. AB - The purpose of the present study was to investigate the bidirectional transport of 1-anilino-8-naphthalene sulfonate (ANS) using isolated rat hepatocytes. The initial uptake rate of ANS by isolated hepatocytes was determined. The uptake process of ANS was saturable, with a Km of 29.1 +/- 3.2 microM and Vmax of 2.9 +/ 0.1 mmol/min/mg protein. Subsequently, the initial efflux rate of ANS from isolated hepatocytes was determined by resuspending preloaded cells to 3.0% (w/v) BSA buffer. The efflux process for total ANS revealed a little saturability. The mean value of the efflux clearance was 2.2 +/- 0.1 microL/min/mg protein. The efflux rate of ANS from hepatocytes was markedly decreased at 4 degrees C, indicating that the apparent efflux of ANS might not be attributed to the release of ANS bound to the cell surface, but to the efflux of ANS from intracellular space. The efflux clearance was furthermore corrected for the unbound intracellular ANS concentration on the basis of its binding parameters to cytosol. The relation between efflux rate and unbound ANS concentration was fitted well to the Michaelis-Menten equation with a saturable and a nonsaturable components. The Vmax and Km values were 0.54 mmol/min/mg protein, and 10.0 microM, respectively. Based on the comparison of the ratios of Vmax to Km (Vmax/Km) corresponding to the transport clearance, the influx clearance was two times higher than the efflux clearance. Together with our preliminary studies that ATP suppression in hepatocytes substantially inhibited ANS influx rate, we concluded that the hepatic uptake of ANS is actively taken up into hepatocytes via the carrier mediated transport system. PMID- 12735693 TI - Effect of excipients on the stability and transport of recombinant human epidermal growth factor (rhEGF) across Caco-2 cell monolayers. AB - The effect of sixteen excipients on the transport of recombinant human epidermal growth factor (rhEGF) across Caco-2 cell monolayers was examined at 37 degrees C. The apparent apical to basolateral (A-B) permeability (Papp) of 30 microM rhEGF was 8.15 x 10(-7) cm/sec, indicative of a poor level of absorption in the GI tract. The Papp was 1.7- and 6.3-fold greater than the Papp in the basolateral to apical (B-A) direction and the A-B permeability of mannitol, respectively, and decreased dramatically to a negligible level at 4 degrees C, consistent with a receptor mediated transcytosis of rhEGF. The stability of rhEGF was very poor, undergoing more than 85% degradation in 2 h in the transport medium at 37 degrees C. A significant increase in the Papp could be achieved by the addition of certain excipients, as exemplified by 23, 21, 20 and 16-fold increases, in the presence of sodium taurochenodeoxycholate (NaTCDC), sodium taurodeoxycholate (NaTDC), sodium glycodeoxycholate (NaGDC) and sodium laurylsulfate (SLS) (all at a concentration of 1% w/v), respectively. A significant increase in stability could also be achieved by the addition of some of the excipients, as represented by 1% SLS, which nearly completely stabilized the rhEGF. Unfortunately, however, an increase in the Papp of rhEGF could not be achieved without a simultaneous and extensive decrease in the integrity of the cell membranes. Thus, more efficient excipients, that specifically enhance the permeation of rhEGF and do not alter the membrane integrity, should be pursued in order to safely enhance the permeation of rhEGF. PMID- 12735695 TI - Diversity of the biosynthesis of the isoprene units. AB - This review covers the biosynthesis of the starter units of terpenoids, isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) via the nonmevalonate pathway together with a new enzyme involved in the conversion of IPP and DMAPP, i.e type 2 IPP isomerase. The biosynthesis of terpenoids produced by actinomycetes is also reviewed. 117 references are cited. PMID- 12735696 TI - Current mechanistic understanding of thiamin diphosphate-dependent enzymatic reactions. AB - The mechanism of thiamin diphosphate-dependent enzymatic reactions is discussed, concentrating on two enzymes involved in decarboxylating pyruvic acid, the yeast pyruvate decarboxylase and the pyruvate dehydrogenase multienzyme complex from Escherichia coli. The availability of high-resolution X-ray structures for several thiamin diphosphate-dependent enzymes, the use of site-specifically substituted protein variants (resulting from site-directed mutagenesis), the development of model reactions for the various putative intermediates, and the application of new mechanistic tools in solution have all contributed to a much better understanding of the role of the protein component in catalysis. Perhaps the most important advance in our understanding of these mechanisms concerns the role of the 4'-aminopyrimidine component of the coenzyme, widely ignored prior to the publication of the X-ray results. The current view is that the two aromatic rings both contribute to catalysis, perhaps carrying out an intramolecular proton transfer to initiate the various reactions, an ability that makes this coenzyme virtually unique among coenzymes. PMID- 12735697 TI - Protein and peptide identification: the role of mass spectrometry in proteomics. AB - Proteomics encompasses several disciplines in the quest to unravel the science of life. Mass spectrometry plays a key role in proteomics-associated analyses and this review describes that function in terms of the instrumentation used, the results generated, data interpretation and the advances in analytical techniques being made to deal with the production of highly specific, high throughput data. The review has 151 references. PMID- 12735699 TI - Aldose reductase inhibitors from natural sources. AB - This review covers aldose reductase inhibitors (ARIs) isolated from natural sources. Compounds in the review are grouped according to the source from which they have been isolated: terrestrial, marine, or microorganism and the in vitro inhibitory activity of the compounds is also showed. The literature, both journals and patents, up to June 2002 is reviewed and 86 references are cited. PMID- 12735698 TI - Simple indole alkaloids and those with a nonrearranged monoterpenoid unit. AB - This review covers simple indole alkaloids and those with a nonrearranged monoterpenoid unit. Newly isolated alkaloids, structure determinations, total syntheses and biological activities are included. The literature from January to December 2001 is reviewed and 166 references are cited. PMID- 12735700 TI - Recent advances in antimicrobial nucleoside antibiotics targeting cell wall biosynthesis. AB - The biosynthesis of peptidoglycan is essential for all bacteria and has no counterpart in eukaryotic cells. It is one of the prime targets for antibiotic chemotherapy, especially phospho-MurNAc-pentapeptide translocase (translocase I) is a fascinating target in which there is no commercial antibiotic. In this review we will describe three nucleoside translocase I inhibitors, mureidomycin, tunicamycin and liposidomycin. PMID- 12735701 TI - Comparison of the standard AGID test and competitive ELISA for detecting bluetongue virus antibodies in camels in Gujarat, India. AB - The performance of the standard agar gel immunodiffusion (AGID) test and the competitive enzyme-linked immunosorbent assay (cELISA) for the detection of serum antibody against bluetongue virus (BTV) in clinically healthy and diseased camels in Gujarat state were compared. Out of 176 sera tested, 22 (12.5%) and 34 (19.3%) were positive for group-specific bluetongue antibodies by AGID and cELISA, respectively. Maximum seropositivities of 18.0% by AGID and 25.8% by cELISA were recorded in the Kutchhi breed, and of 6.9% and 12.6%, respectively, in the Marwari breed. The seroprevalence detected by AGID and cELISA in clinically healthy and diseased camels did not differ significantly with regard to bluetongue disease in these breeds. PMID- 12735702 TI - Dairy production and reproduction by crosses of Holstein, Jersey or Brown Swiss sires with Holstein-Friesian/Gir dams. AB - The performance was compared of cows sired by Holstein, Jersey or Brown Swiss bulls out of Holstein-Friesian x Gir dams of 1/2 to 3/4 Holstein-Friesian content. The animals were kept in a single herd under the same management. The data were analysed by least-squares techniques under a model that included the fixed effects of breed of sire. Bos taurus fraction of the dam, parity, year and season of calving, and a random cow effect. Based on 480 observations, the milk yields per lactation for the Holstein, Jersey and Brown Swiss sired groups were 2,821 +/- 163, 2,320 +/- 61 and 2,418 +/- 119 kg, respectively. The corresponding means for fat yield per lactation were 96.9 +/- 6.6, 86.8 +/- 2.5 and 92.8 +/- 4.8 kg; for protein yield per lactation were 85.3 +/- 5.1, 71.3+/-1.9 and 76.3 +/ 3.7 kg; for lactation length, 339 +/- 18, 283 +/- 7 and 313 +/- 14 days for fat percentage. 3.37 +/- 0.10, 3.73 +/- 0.04 and 3.77 +/- 0.07%; and for protein percentage, 3.02 +/- 0.05, 3.10 +/- 0.02 and 3.16 +/- 0.04%. The respective calving intervals were 487 +/- 24, 408 +/- 11 and 461 +/- 245 days. The yields of milk and protein per day of calving interval were similar in the Jersey and Holstein sired groups, while the former had higher yields of fat, implying that production economics might favour the smaller Jersey crosses in production systems in which the males were not reared. Crossing with Brown Swiss did not improve performance over the Holstein sired crosses. PMID- 12735703 TI - Reproduction and lactational performance of cattle in a smallholder dairy system in Zimbabwe. AB - A study was conducted in two adjacent locations. Nharira (communal) and Lancashire (small-scale commercial) farming areas in Zimbabwe to characterize the breeds and evaluate the reproductive and lactation performance of dairy cattle under smallholder management. The types of cows identified were Friesian, Jersey and Red Dane, and an indigenous Sanga breed called the Mashona and its crossbreds. Both sectors used more exotic and crossbred cows than indigenous cows. The mean monthly weights of the dairy cows were higher in Lancashire than in Nharira and the calving intervals were longer in Nharira than in Lancashire. The mean age at first calving was higher and the mean total lactation yields were greater in Nharira than in Lancashire, but the mean 305-day lactation yields were not significantly different. The mean lactation lengths were longer for the cows from Nharira. It was concluded that the reproductive and lactation performances were low. The calving intervals were extended, probably owing to suboptimal nutrition and heat stress, particularly during the dry season, and to poor management practices, such as delayed mating due to the poor availability of bulls. PMID- 12735704 TI - Replacement value of undecorticated sunflower meal as a supplement for milk production by crossbred cows and buffaloes in the northern plains of India. AB - The replacement value of undecorticated sunflower meal (SFM) in the diets of dairy animals was assessed on-station and on-farm. Eighteen primiparous crossbred (Bos taurus x Bos indicus) cows (350.4 +/- 8.84 kg), randomly allocated to three groups, were used in the on-station study. The animals were fed on either a conventional concentrate supplement (control) or on an experimental concentrate, in which SFM replaced 25% (SFM-25) or 50% (SFM-50) of the CP in the control supplement. Green oats (Avena sativa) were supplied ad libitum. A metabolism trial conducted following 60 days of experimental feeding revealed that the intakes of DM, DCP and TDN were similar among the groups. The digestibilities of OM, CP, EE, NDF and ADF were also without significant differences. All the groups were in positive nitrogen balance. Inclusion of SFM at either level had no effect on the intake, excretion or retention of nitrogen. The daily milk yield and its composition did not differ among the dietary treatments. Moreover, the efficiency of utilization of DOM and TDN for FCM production tended to reflect, although non significantly, increasing levels of SFM inclusion. In the on-farm study, seven multiparous milking buffaloes belonging to six farmers were used to assess the effect of replacing 20% of the CP of the conventional supplement with SFM, in a predominantly crop residue-based diet. The study continued for 4 months and revealed that the average daily feed intake and milk production was similar in the control and SFM-fed groups. It was concluded that SFM can be effectively utilized as a cheaper replacement for costly oil cakes and wheat bran for economic milk production by smallholder farmers. PMID- 12735705 TI - Milk protein polymorphism in Kangayam cattle. AB - This paper reports the milk protein polymorphism, the allele frequencies of variants and the possible linkages among various combinations of milk protein phenotypes in the Kangayam cattle of south India. Milk samples from 156 Kangayam cows were typed by starch gel and polyacrylamide gel electrophoresis for caseins and whey proteins, respectively. All the four milk protein components studied, alpha(s)1-casein. beta-casein, beta-lactoglobulin and alpha-lactalbumin, exhibited polymorphism with high allele frequencies of 0.9231 +/- 0.0151 for alpha(s)1-casein C, 0.9263 +/- 0.0148 for beta-casein A, 0.9135 +/- 0.0159 for beta-lactoglobulin B and a relatively high frequency of 0.6218 +/- 0.0275 for alpha-lactalbumin A. The mean heterozygosity estimated over all the four milk protein loci was 0.2420. Genetic equilibrium was observed among all the loci studied, except alpha-lactalbumin. Linkage analysis confirmed the non independence between alpha(s)1- and beta-caseins and between caseins and alpha lactalbumin phenotypes. PMID- 12735707 TI - Energy supply to livestock from tropical rangeland during the dry season. AB - Nine key forage species (grasses and legumes), together with two types of crop residues, usually fed by farmers to their livestock, were collected from a rainfed area in western Sudan during the dry season (May-April). The grasses investigated were Leptadena pyrotechnia, Cenchrus setigrus, Arista pallida, Eragrotis tremula, Schoenefeldia gracilis, Chloris vergata and Cenchrus biflorus. The crop residues investigated were the grasses, sorghum straw (Sorghum bichlor) and millet straw (Pennisetum typhodium) and the legumes Stylosanthes flavicans and Cajanus cajana. Estimates of organic matter (OM) degradability were done using the nylon bag technique, which was fitted into the model Y = a + b (1 - e( ct)), in which the asymptote (a + b) represented the total potential degradability. Organic cell wall constituents and hence both metabolizable energy and total digestible energy or nutrients (TDN) were determined. S. flavicans showed the best organic matter degradability, and sorghum straw was better degraded than millet straw. The rest of the grasses showed poor OM degradability. Acid detergent insoluble nitrogen was inversely related to TDN, the latter falling within a narrow range for the different forages. Fermentable metabolizable energy differed only slightly, while the legume S. flavicans had the highest effective rumen digestible protein. Undegraded proteins were high for the straws and the grasses L. pyrotechnia and C. setigerus. Metabolizable protein and microbial protein were highest in the sorghum straw, C. setigerus and S. flavicans. PMID- 12735706 TI - An in vitro evaluation of some unconventional ruminant feeds in terms of the organic matter digestibility, energy and microbial biomass. AB - In vitro organic matter apparent digestibility (IVOMAD), true digestibility (IVOMTD), metabolizable energy (ME), net energy lactation (NEL), microbial nitrogen (MN) and synthesis of microbial biomass (MBM) were estimated to predict the nutritive values of some agricultural by-products, drought-tolerant range plants and browses. The relationships between in vitro gas production (GP), and true or apparent digestibility. MN and MBM were studied utilizing an in vitro incubation technique. The values of IVOMAD, IVOMTD, ME, NEL, GP, MBM and MN varied with the studied experimental materials. The true fermentation of the outside part of Atriplex leucoclada produced a higher volume of gas than the middle or the inside parts, and this was associated with an increase in the values of IVOMAD, IVOMTD, ME and NEL. However, screening off the wood from olive cake to obtain olive cake pulp increased the IVOMAD, IVOMTD, ME, NEL and the volume of gas production from the true fermented material. One ml of gas was generated from the true degradation of 5 mg of wheat straw, Moringa oleifera, Alhagi camelorum, Eucaliptus camaldulensis and A. leucoclada, from 11 mg of Prosopsis stephaniana and olive cake pulp, and from 20 mg of olive cake or olive cake wood. The amount of MN or MBM produced from 100 mg of truly fermented organic matter depended on the kind of the fermented material and amounted to 0.7 2.9 mg or 8-34 mg, respectively. Crude fibre was negatively correlated to IVOMAD, IVOMTD, ME and NEL. Gas production was positively correlated to IVOMAD and IVOMTD but negatively correlated to MBM and MN. PMID- 12735708 TI - Ownership pattern and management practices of cattle herds in the Gambia: implications for a breeding programme. AB - A questionnaire and a census were carried out in 1998/99 on 201 cattle herds in 44 villages located in three different areas of The Gambia, in order to obtain information about the ownership pattern, management and breeding practices for optimizing and extending a pure breeding scheme for indigenous N'Dama cattle. The herds had an average of 58.5 head, of which 39.3% were cows, and most of them were multiowner herds, having on average 5.5 owners. Overall, women represented 38.3% of the owners, but they owned only 15.9% of the cattle. Some of the senior herdsmen received no payment, as they were family members, but most of them received payment in kind, i.e. milk off-take from the herd. Payment in cash only or cash as well as milk was rare. In larger herds it was quite common for additional herdsmen to be hired throughout the year, their payment shifting towards cash only or cash and milk. Many small herds had no older bulls or breeding bull and almost all breeding bulls were born within the herd. In most cases, it was the herd owner who was responsible for the selection of the breeding bull. Traits related to the growth and milk production of the dam were considered as most important for a good breeding bull. The results are discussed with respect to the extension of a pure breeding scheme for N'Dama cattle. PMID- 12735709 TI - Growth and apparent absorption of minerals in broiler chicks fed diets with low or high tannin contents. AB - Sorghum grains with low (0.28 per cent catechin equivalent (%CE) or high (1.36%/DCE)) tannin contents were used to study the effect of tannin on growth and on the apparent absorption of Ca, P, Mg, Na, K, Fe and Co by chickens. High tannin sorghum caused a highly significant (p < or = 0.01) reduction in the weight gain and feed intake of broiler chicks compared to low-tannin sorghum and increased the feed conversion ratio (p < or = 0.01). The amount of tannin did not affect mortality. The apparent absorption of Ca, P, Mg, Na, K, Fe and Co was reduced (p < or = 0.01) by feeding sorghum grains with a high tannin content. PMID- 12735710 TI - Public health and child maltreatment prevention: the role of the Centers for Disease Control and Prevention. AB - Child maltreatment is a serious and preventable public health problem. Recent studies indicate a dose-response relationship between exposure to child maltreatment and the presence of adult diseases, clearly positioning child maltreatment as a public health burden. This commentary describes the Center for Disease Control and Prevention's (CDC) public health approach to prevention, and identifies elements of the CDC role that are complementary to the efforts of criminal justice and child protective services. CDC's goal for child maltreatment prevention is to assure the widespread adoption of prevention and intervention strategies that are evidence based. Immediate and practical impact can occur by improving collection of child maltreatment data, emphasizing positive parenting skills, and promoting programs representing the best prevention practices in child maltreatment prevention. PMID- 12735711 TI - A multilevel study of neighborhoods and parent-to-child physical aggression: results from the project on human development in Chicago neighborhoods. AB - The majority of children in the United States experience parent-to-child physical aggression (PCPA), a disciplinary strategy out of favor with many experts. Several decades of research have documented a link between community characteristics and severe child maltreatment. None have taken a multilevel approach to study whether neighborhoods affect the amount of corporal punishment and/or physical abuse used by individual families. Data for this article come from the Project on Human Development in Chicago Neighborhoods and were analyzed using hierarchical linear modeling. An interval scale of PCPA was developed. Values obtained show that several neighborhood characteristics were associated with PCPA. Immigrant concentration remained significant after controlling for family composition. A cross-level interaction was found between neighborhood social networks and Hispanic race/ethnicity. The article's conclusion is that neighborhood characteristics may influence the amount of PCPA used by families. Neighborhood intervention strategies hold promise. PMID- 12735712 TI - Improving the measurement of child neglect for survey research: issues and recommendations. AB - There is a great need for developing and validating measures of child neglect that can be applied to survey samples outside of a child welfare context. A prospective assessment of child neglect would afford a better estimation of the etiology of various types of child neglect and would greatly inform the development of primary prevention strategies related to child maltreatment. This article offers guidance on the tasks involved with constructing new measures of neglect for prospective survey research. Methodological issues pertaining to child neglect measurement are discussed, and a framework is offered for developing neglect measures for survey research. A discussion is also offered on how this framework is being applied in an ongoing longitudinal study of low income families with young children. The intended result of this exercise is to encourage the development of new child neglect measures for survey research with both high-risk and general populations. PMID- 12735713 TI - Decomposing Black-White differences in child maltreatment. AB - This article examines conflicting visions of the racial composition of the maltreated populations. The National Child Abuse and Neglect Data System (NCANDS) data shows Blacks are overrepresented among reported and substantiated abuse and neglect cases, while the National Incidence Study of Child Abuse and Neglect (NIS) shows no apparent overrepresentation of children of color. To understand the conflicting evidence, the authors produce from NIS approximate measures of maltreatment rates in NCANDS. Maltreatment rates is broken down into allegation, report and substantiation components. Without disaggregating the data by welfare status, all or most of the racial gap in official maltreatment is found to arise from racial differences in allegations. Disaggregation changes the results. Among welfare cases, on average, half of the Black-White gap in maltreatment is due to racial differences in substantiation. Among nonwelfare cases, about half of the racial gap in official maltreatment is due to racial differences in allegations. PMID- 12735714 TI - Adolescent pregnancy and homicide: findings from the Maryland Office of the Chief Medical Examiner, 1994-1998. AB - Homicide has remained the third leading cause of death among girls aged 10 to 19 for more than a decade. Recent research indicates that pregnant or postpartum teens are three times more likely to be victims of homicide compared to their nonpregnant counterparts. These findings portray a compelling picture that leads the authors to investigate the relationship between homicide and pregnancy among teens in Maryland. The purpose of this study is to compare women whose deaths had been evaluated by the medical examiner and who had evidence of pregnancy to women without evidence of pregnancy, with a particular focus on adolescents; 329 (17.7%) adults and 66 (32.7%) adolescents were victims of homicide. Adolescent homicide victims were 3.7 (1.2 to 11.8) times more likely to be pregnant compared to adult homicide victims. The rate of homicide was nearly double in all women who were pregnant. Further research is necessary to evaluate factors associated with these risks so that the future generation and society are protected. PMID- 12735715 TI - The effect of childhood maltreatment on adult criminality: a tobit regression analysis. AB - This article reports on the results of an analysis of a data set containing information on 667 nonmaltreated and 908 maltreated children. The data also contain information on whether the study subjects were arrested in early adulthood. Because adult arrests are an imperfect and censored measure of antisocial behavior, tobit regression analysis was used to examine the effect of the subjects' experiences of child maltreatment on later arrests while controlling for those subjects' demographic characteristics. The analysis finds that children's age, race, and sex and experiences of child neglect all have an impact on subsequent adult arrests. However, physical abuse and sexual abuse do not emerge as statistically significant predictors of arrests in this model. The study also illustrates a method for the decomposition of tobit coefficients to extract more information from them. PMID- 12735716 TI - Medical, legal, and mental health service utilization by physically abused children and their caregivers. AB - The use of medical, legal, and mental health services was examined among 37 Medicaid-eligible, physically abused children and their caregivers. Fewer than half of the children received a medical examination related to the physical abuse, but the majority had received basic wellness care, including immunizations. Roughly half of the cases were heard in family court. Fewer than half of the children were receiving mental health services. Children were more likely to receive services if the maltreating caregiver was not in the home. More maltreating caregivers received mental health services than did their children, but this still only accounted for half of the parents. Caregivers were more likely to receive treatment if they acknowledged the abuse. Children who participated in treatment showed reductions in parent-reported problem behaviors but showed increases in anxiety in comparison to children who did not participate in treatment. PMID- 12735717 TI - Defining and classifying supervisory neglect. AB - The purpose of this study is to develop and test a classification system for different types of supervision problems. Several prominent child maltreatment typologies were reviewed to determine the level of definitional agreement between systems; identify strengths and weaknesses within systems; and derive a consensual definition of child neglect, its major subcategories (physical, supervisory, and emotional), and 10 types of supervisory neglect. Six hundred two substantiated child abuse and neglect reports were used to test the systems interrater reliability. The interrater reliability was excellent for most types of problems. Not watching a child closely enough (29.8% of the reports) made up the largest single type of supervisory neglect, followed by leaving a child alone without a caretaker (24.5%) and leaving a child with an unsuitable caretaker (19.7%). The article concludes with a discussion on how and in what context the system can be used. PMID- 12735718 TI - Spermidine metabolism in parasitic protozoa--a comparison to the situation in prokaryotes, viruses, plants and fungi. AB - Targeting polyamines of parasitic protozoa in chemotherapy has attracted attention because polyamines might reveal novel drug targets for antiparasite therapies (Muller et al. 2001). The biological function of the triamine spermidine in parasitic protozoa has not been studied in great detail although the results obtained mainly imply three different functions, i.e., cell proliferation, cell differentiation, and biosynthesis of macromolecules. Sequence information from the malaria genome project databases and inhibitor studies provide evidence that the current status of spermidine research has to be extended since enzymes of spermidine metabolism are present in the parasite (Kaiser et al. 2001). Isolation and characterisation of these enzymes, i.e., deoxyhypusine synthase (EC 1.1.1.249) (DHS) and homospermidine synthase (EC 2.5.1.44) (HSS) might lead to valuable new targets in drug therapy. Currently research on spermidine metabolism is based on the deposition of the deoxyhypusine synthase nucleic acid sequence in GenBank while the activity of homospermidine synthase was deduced from inhibitor studies. Spermidine biosynthesis is catalyzed by spermidine synthase (EC 2.5.1.16) which transfers an aminopropyl moiety from decarboxylated S-adenosylmethionine to putrescine. Spermidine is also an important precursor in the biosynthesis of the unusual amino acid hypusine (Wolff et al. 1995) and the uncommon triamine homospermidine in eukaryotes, in particular in pyrrolizidine alkaloid-producing plants (Ober and Hartmann 2000). Hypusine is formed by a two-step enzymatic mechanism starting with the transfer of an aminobutyl moiety from spermidine to the epsilon-amino group of one of the lysine residues in the precursor protein of eukaryotic initiation factor eIF5A by DHS (Lee and Park 2000). The second step of hypusinylation is completed by deoxyhypusine hydroxylase (EC 1.14.9929) (Abbruzzese et al. 1985). Homospermidine formation in eukaryotes parallels deoxyhypusine formation in the way that in an NAD(+)-dependent reaction an aminobutyl moiety is transferred from spermidine. In the case of homospermidine synthase, however the acceptor is putrescine. Thus the triamine homospermidine consists of two symmetric aminobutyl moieties while there is one aminobutyl and one aminopropyl moiety present in spermidine. Here, we review the metabolism of the triamine spermidine with particular focus on the biosynthesis of hypusine and homospermidine in parasitic protozoa, i.e., Plasmodium, Trypanosoma and Leishmania, compared to that in prokaryotes i.e., Escherichia coli, a phytopathogenic virus and pyrrolizidine alkaloid-producing plants (Asteraceae) and fungi. PMID- 12735719 TI - Long-lasting anticryptosporidial activity of nitazoxanide in an immunosuppressed rat model. AB - Cryptosporidium parvum, Tyzzer, 1912 is identified as a common cause of diarrhoea in immunocompetent individuals. In immunocompromised, especially HIV-infected subjects, cryptosporidiosis causes severe chronic diarrhoea. In this study, nitazoxanide (NTZ) was compared for curative activity with sinefungin (SNF) and paromomycin (PRM) in immunosuppressed rats, a screening model for anticryptosporidial agents. NTZ at either 50 mg/kg/day, 100 mg/kg/day or 200 mg/kg/day resulted in seven days in a dose-dependent inhibition of oocyst shedding similar to that obtained with SNF (10 mg/kg/day) and PRM (100 mg/kg/day). Further discontinuation of SNF or PRM 100 mg/kg/day therapy resulted in early relapse of oocyst shedding which reached the pre-treatment levels in 2-4 days. In contrast, seven days after discontinuation of therapy, shedding inhibition was unchanged in NTZ-treated rats. Data prompt further assessment of the activity of NTZ on sequestered C. parvum. PMID- 12735720 TI - Description of Eimeria arabukosokokensis sp. n. (Apicomplexa: eimeriidae) from Telescopus semiannulatus (Serpentes: Colubridae) with notes on eimerian coccidia from snakes of Eastern Kenya. AB - Parasitological examination of faeces of 26 snakes kept in Bio-Ken Snake Farm, Watamu, Kenya revealed new species of Eimeria Schneider, 1875 in Telescopus semiannulatus Smith, 1849. Oocysts of Eimeria arabukosokokensis sp. n. are cylindrical 26.8 (25-29) x 15.1 (14-16) microm with smooth, bilayered oocyst wall and a single polar granule. The broadly ellipsoidal sporocysts average 9.3 (8.5 10) x 7.1 (6.5-7.5) microm and possess single-layered wall composed of two plates joined by longitudinal suture. Caryospora cf. regentensis Daszak et Ball, 2001 is reported from Dendroaspis angusticeps (Smith, 1849) and two additional forms of Caryospora Leger, 1904 are reported and morphologically characterised from a single specimen of Psammophis orientalis Broadley, 1977. Systematic status of Caryospora spp. in sub-Saharan Psammophis Boie, 1827 is discusses and all species reported by various authors to date are suggested to be treated as species inquirendae until more detailed data on these parasites and their hosts are available. PMID- 12735721 TI - Infection in the fins of the goldfish Carassius auratus caused by Myxobolus diversus (Myxosporea). AB - During health surveys of 7- to 9-week-old goldfish (Carassius auratus L.) fingerlings in a fish farm near Budapest, Hungary, myxosporean plasmodia were observed on the fins. Plasmodia were most frequently found at the intersegmental joints of the finrays. Spores of Myxobolus diversus Nie et Li, 1973 known from China, were detectable in the mature plasmodia located within the lumen, and less often on the surface, of the cartilaginous finrays. The external wall of the plasmodia was constituted by a capsule formed from a collagenous material identical with the cartilaginous substance of the finrays. Matured plasmodia were filled by spores of 12-14 x 8-9.5 microm in size. The relatively small plasmodia caused only small deformities on the fins. Their importance is, however, not negligible, as in an ornamental fish such as the goldfish even a minor damage of the fins causes a loss of value. Besides a report on the first European occurrence and pathological aspects, a redescription of this parasite of Far Eastern origin is given. PMID- 12735722 TI - Fine structure of Henneguya pilosa sp. n. (Myxozoa: Myxosporea), parasite of Serrasalmus altuvei (Characidae), in Brazil. AB - Henneguya pilosa sp. n., a new species of myxosporean from the gill filaments of the white piranha, Serrasalmus altuvei Ramirez, 1965 (Characidae), a freshwater teleost fish collected in the Zoological Garden of the city of Teresina (Piaui), Brazil, is described from light and transmission electron microscope observations. This myxosporean produced small plasmodia (up to 0.2 mm in diameter), each one containing all life-cycle stages of the parasite, including numerous spores. The spores, laterally compressed, averaged 54.2 (52.3-56.0) microm in total length and consisted of two unequal valves adhering together along the suture line and two caudal processes. The spore body measured 21.1 (20.0-23.1) microm in length, 5.9 (5.5-6.3) microm in width, and 2.2 (1.9-2.6) microm in thickness. The two equal ellipsoidal polar capsules of 7.4 (7.1-7.6) microm long and 1.2 (1.0-1.3) microm wide possessed a polar filament with 11-12 (rarely 13) turns. All surfaces of the spores were covered with a tightly adherent complex network of numerous densely ramified granulo-fibrillar masses, the longest measuring 1.5 microm long, observed around the caudal processes. The prevalence of infection was 30%. The taxonomic affinities of this parasite with other of the same genus in freshwater South American fish species are discussed. PMID- 12735723 TI - Pseudoallencotyla gen. n., a new genus for Allencotyla pricei (Kritsky, Noble et Moser, 1978) (Monogenea: Heteraxinidae), with a key to genera of Heteraxininae. AB - In this study, we clarify the taxonomic status of Allencotyla pricei based on a re-examination of paratypes and voucher specimens. Four important characters are added to previous descriptions: the vagina is ventral; the cirrus is absent; the eggs are fusiform and have two polar filaments (one about 3.5 times longer than the other); and, there are two flat sclerites at each side of the dorsal trident shaped sclerite in the clamps. A comparison of A. pricei with species from all the genera of the Heteraxininae, including Allencotyla mcintoshi, indicates that A. pricei has several exclusive characters, i.e., a ventral vagina, vaginal armature, two additional dorsal sclerites in the clamps, a ventral mouth, and a question mark-shaped ovary. The combination of characters found in A. pricei is not shared with any other species within the Heteraxininae. In view of these features, A. pricei is assigned to a new genus, Pseudoallencotyla gen. n., as P. pricei comb. n. PMID- 12735724 TI - Mormyrogyrodactylus gemini gen. et sp. n. (Monogenea: Gyrodactylidae), a new gyrodactylid from Marcusenius macrolepidotus (Mormyridae) from South Africa. AB - Mormyrogyrodactylus gemini gen. et sp. n. (Gyrodactylidea: Gyrodactylidae), a viviparous monogenean is described from the skin and fins of Marcusenius macrolepidotus (Peters) from South Africa. This new genus is unique in having a large cup-shaped cirrus armed with one large needle-like spine and numerous small hair-like spinelets. A peduncular bar is present. The haptor is clearly demarcated from the body and situated on a small pedicle. The haptor has one pair of large anchors, a ventral bar complex, a small dorsal bar and 16 evenly spaced marginal hooks. The ventral bar complex consists of three parts: an inverted U shaped piece with two semi-attached bars, each bar consisting of a base and an extension. An accessory inverted T-shaped sclerite is present on the posterior rim of the haptor. Mormyrogyrodactylus is the fifth genus of the Gyrodactylidae to be described from Africa. PMID- 12735725 TI - Toxicity of cadmium and zinc to the cercarial activity of Diplostomum spathaceum (Trematoda: Diplostomidae). AB - The toxicity of cadmium and zinc at concentrations ranging from 0.1 to 100 microg/l was investigated against the activity of Diplostomum spathaceum (Rudolphi, 1819) cercariae. Over a 24 h exposure period a significant reduction in cercarial activity occurred in solutions of cadmium, zinc, and a mixture of cadmium and zinc at all concentrations. Reduced cercarial activity also occurred in all toxicant solutions compared with controls after only 6 h exposure indicating that cercariae were vulnerable during the period of maximum cercarial infectivity (0-5 h). The mechanisms of metal toxicity and their importance to parasite transmission are discussed. PMID- 12735726 TI - Review of the Rhopalothylacidae Guiart, 1935 (Cestoda: Trypanorhyncha), with a description of the adult of Pintneriella musculicola Yamaguti, 1934 and a redescription of P. gymnorhynchoides (Guiart, 1935) comb. n. AB - The family Rhopalothylacidae (Cestoda: Trypanorhyncha) is reviewed. The type species, Rhopalothylax gymnorhynchoides Guiart, 1935, is redescribed from the type specimens and belongs within the genus Pintneriella Yamaguti, 1934, previously described only from the plerocercus. Rhopalothylax therefore becomes a junior synonym of Pintneriella. The adult of Pintneriella musculicola Yamaguti, 1934 is described for the first time, from the shark Carcharias taurus Rafinesque from Australia. Pintneriella is characterised by two bothridia, a typical heteroacanthous armature, a unique, bipartite external seminal vesicle and a uterus deviated porally, terminating at a uterine pore. It belongs within the Heteracanthoidea but is distinguishable both from the Eutetrarhynchidae and the Gilquiniidae, the two families which it most closely resembles. Cladistic analyses align Pintneriella within the clade containing the families Gilquiniidae, Gymnorhynchidae and Molicolidae rather than with the Eutetrarhynchidae. The family Rhopalothylacidae is therefore retained provisionally to accommodate Pintneriella within the Heteracanthoidea. The second genus of the Rhopalothylacidae, Clujia Guiart, 1935, is unrecognisable from its description and cannot be redescribed from its holotype. It is therefore considered a genus inquirendum. PMID- 12735727 TI - Component population study of Acanthocephalus tumescens (Acanthocephala) in fishes from Lake Moreno, Argentina. AB - Seasonal samples of all fish species from Lake Moreno were taken in order to determine the presence of paratenia, to evaluate the status of the hosts and to characterise the transmission of Acanthocephalus tumescens (von Linstow, 1896) at the component population level. Prevalence, mean abundance, mean intensity, numbers of gravid females, relative abundance of the different fish species, relative output of eggs and relative flow rates for each host species were computed. Acanthocephalus tumescens showed low host specificity, successfully parasitizing six out of eight fish species present in the lake. No paratenic infection was registered. If prevalence, mean abundance, and number of gravid females are considered, host species can be placed in a continuum from the most to least suitable as follows: Galaxias platei Steindachner, Diplomystes viedmensis (Mac Donagh), Oncorhynchus mykiss (Walbaum), Salvelinus fontinalis (Mitchill), Percichthys trucha (Cuvier et Valenciennes) and Galaxias maculatus (Jenyns). However, when parasite flow rates and egg output were calculated, including relative abundance of each fish species, the continuum was rearranged as follows: P. trucha, O. mykiss, G. platei / G. maculatus, S. fontinalis and D. viedmensis. The first four species would be the main contributors to the population of A. tumescens in this lake, P. trucha being the major one. Different regulatory and non-regulatory mechanisms are suggested. PMID- 12735729 TI - Observations on Babesiosoma mariae (Apicomplexa: Dactylosomatidae) from the Okavango Delta, Botswana. PMID- 12735728 TI - Effect of fast protein liquid chromatography fractionated salivary gland extracts from different ixodid tick species on interleukin-8 binding to its cell receptors. AB - Interleukin-8 plays a critical role in inflammatory processes. Hence generation of molecules with anti-IL-8 activity is likely to be important for successful feeding and for survival of the ticks. Anti-IL-8 activity was studied in saliva of three ixodid tick species--Dermacentor reticulatus (Fabricius, 1794), Rhipicephalus appendiculatus Neumann, 1901, and Amblyomma variegatum (Fabricius, 1794). The greatest activity was shown in saliva prepared from D. reticulatus. The activity was attributed to tick salivary gland molecules that bind to IL-8, preventing binding of the chemokine to its specific receptor, rather than to occupation of the IL-8 cell receptor by the tick molecules. The distribution of anti-IL-8 activity in fast protein liquid chromatography (FPLC) fractions of salivary gland extracts (SGE) derived from adult female D. reticulatus, R. appendiculatus and A. variegatum was compared directly by both ELISA and receptor binding inhibition assays. The correspondence in results with fractions of SGE from ELISA is consistent with detection of tick molecules that inhibit IL-8 binding to its receptor. As IL-8 is an important chemoattractant and activator of neutrophils, the presence of an anti-IL-8 activity in tick saliva indicates that neutrophils play an important role in the host response to parasitism by ticks. PMID- 12735730 TI - Highly satisfactory alkynylation of alkenyl halides via Pd-catalyzed cross coupling with alkynylzincs and its critical comparison with the sonogashira alkynylation. AB - [reaction: see text] The Pd-catalyzed alkynylation of various alkenyl halides and triflates with alkynylzincs proceeds well even with alkynyl derivatives containing electron-withdrawing groups. The reaction appears to be highly general. Noteworthy is that the corresponding Sonogashira reactions under various reported conditions are significantly less satisfactory in all cases performed in this study. PMID- 12735731 TI - Efficient chiral discrimination by 77Se NMR. AB - [reaction: see text] Several (77)Se NMR experiments were performed by titrating a sample of selenides with the chiral shift reagent methylbenzylamine (MBA), followed by acquisition of (77)Se NMR spectra. Eventually, we observed the appearance of two anisochronous resonances, with a relatively large separation, from 37 to 56 Hz, corresponding to the formation of the diastereomeric complexes. This methodology avoids derivatization processes, and the studied compound can be easily recovered from the NMR tube. PMID- 12735732 TI - Unprecedented hydrothermal reaction of o-phenylaniline and related derivatives with cyclic ketones. A novel approach to the construction of phenanthridine and quinoline ring systems. AB - [reaction: see text] A new method for synthesizing phenanthridine and its related compounds was developed using the condensation of o-phenylaniline and its homologues with cyclic ketones under hydrothermal conditions. PMID- 12735733 TI - Mercuric triflate catalyzed hydroxylative carbocyclization of 1,6-enynes. AB - [reaction: see text] Hg(OTf)(2) exhibits remarkable catalytic activity for the hydroxylative cyclization of 1,6-enynes. The present procedure should involve a sequence of mercuration of a terminal alkyne, carbocyclization, hydration, and protodemercuration that regenerates the catalyst. PMID- 12735734 TI - Progress toward the total synthesis of (+)-aldosterone: synthesis of the A-D rings. AB - [reaction: see text] Synthesis of the A-D rings of the cortical hormone (+) aldosterone is described. The key step incorporates a chiral tether in a type 2 intramolecular Diels-Alder reaction that establishes the absolute configuration of four contiguous asymmetric centers. This approach provides an efficient route for either enantiomer of the steroid skeleton. PMID- 12735735 TI - Synthetic studies on borrelidin: enantioselective synthesis of the C1-C12 fragment. AB - [structure: see text] An efficient, enantioselective synthesis of the C1-C12 fragment 2 of borrelidin is presented. Construction of the "skipped" polymethylene chain of 2 was accomplished by iteration of Myers' alkylation, while formation of the C3 stereocenter was achieved by Roush's asymmetric allylboration methodology. PMID- 12735736 TI - Asymmetric reductive ring-opening of bicyclic olefins catalyzed by palladium and nickel complexes. AB - [reaction: see text] Asymmetric reductive ring opening of oxa- and azabenzonorbornadienes with organic acids and zinc powder under mild conditions catalyzed by Ni(binap)Cl(2) or Pd(binap)I(2) produces the corresponding 1,2 dihydronaphth-1-ols in good to excellent yields with high enantioselectivity. PMID- 12735737 TI - Remarkably enhanced excimer formation of naphthylacetate in cation-charged gamma cyclodextrin. AB - [reaction: see text] 6(A),6(D)-Bispyridinio-appended gamma-cyclodextrin effectively enhanced the excimer fluorescence of 2-naphthylacetate. This increase derived from the increase of formation of the 1:2 complex between the cation charged gamma-cyclodextrin and 2-naphthylacetate by the electrostatic interaction between the host and the guest. PMID- 12735738 TI - Metabolic engineering to produce sesquiterpenes in yeast. AB - [reaction: see text] Presented here is a metabolically engineered yeast strain that produces sesquiterpenes. Epi-cedrol synthase expressed in a native yeast strain converted endogenous farnesyl pyrophosphate to 90 microg/L epi-cedrol. This system was genetically modified to increase foreign terpene yields to 370 microg/L. The best yields were obtained by overexpressing a truncated 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase in a upc2-1 mating type a background. This system allows sufficient production to characterize novel sesquiterpene synthase genes. PMID- 12735739 TI - S-(2-pyridinyl)-1,1,3,3-tetramethylthiouronium hexafluorophosphate. A new reagent for the synthesis of 2-pyridinethiol esters. AB - [reaction: see text] A new thiouronium-based reagent for the synthesis of 2 pyridinethiol esters under non-nucleophilic conditions from the corresponding carboxylic acids was developed. The resulting procedure enables the preparation of previously unavailable alpha,beta-unsaturated 2-pyridinethiol esters as well as their aliphatic and aromatic counterparts. PMID- 12735740 TI - Highly diastereoselective and asymmetric Diels-Alder reactions of masked o benzoquinones with chiral racemic and homochiral furans. Synthesis of optically active tricyclic gamma-lactones. AB - [reaction: see text] The first examples of highly diastereoselective and asymmetric Diels-Alder cycloadditions of in situ generated masked o-benzoquinones (MOBs) with chiral racemic and homochiral furans bearing a chiral center in the alpha-position leading to highly functionalized diastereomeric/enantioselective tricyclic heterocycles and chiral tricyclic gamma-lactones are described. PMID- 12735741 TI - Molecular recognition of amidines in water. AB - [structure: see text] Tetraphosphonates of the general structure shown above are biomimetic hosts for bisamidinium cations in drugs such as pentamidine and DAPI. Similar to their insertion into DNA's minor groove, these drugs are often sandwiched by two tetraphosphonate hosts (2:1). The alternative binding mode (1:2) produces extremely high association constants in water of approximately 10(8) M(-)(2) ( approximately 12 kcal/mol), which can compete with the biological process. PMID- 12735742 TI - On the reduction of S-alkyl-thionocarbonates (xanthates) with phosphorus compounds. AB - [reaction: see text] Reductive cleavage of the carbon-sulfur bond present in S alkyl-thionocarbonates (xanthates) was achieved by high-yielding, tin-free radical reactions based on phosphorus reagents. The combination hypophosphorous acid/triethylamine/AIBN led to fast, efficient, and smooth formation of the alkane. Reduction with diethyl phosphite was sufficiently slow to permit sequential intermolecular addition of a 2-oxoalkyl xanthate onto an olefin followed by cleavage of the newly formed carbon-sulfur bond. PMID- 12735743 TI - Stereoselective entry to beta-linked C-disaccharides using a carbon-Ferrier reaction. AB - [structure: see text] The synthesis of unsaturated beta-linked C-disaccharides by the Lewis acid-mediated reaction of 3-O-acetylated glycals with monosaccharide derived alkenes is described. Deprotection and selective hydrogenation of an exocyclic carbon-carbon double, in the presence of an endocyclic double bond, for representative targets is also illustrated. PMID- 12735744 TI - Mild procedure for the catalytic bis(stannylation) of alkynes with hexaalkylditins. AB - [reaction: see text] Bis(stannylation) of terminal alkynes is achieved through the use of a palladium-isonitrile catalyst complex using a hexaalkylditin as a stannyl group transfer reagent in an atom-efficient and mild catalytic process. Functional group tolerance is good, allowing the presence of amine, carbamate, silyl, ester, and ether moieties. An activated internal alkyne also underwent bis(stannylation) in moderate yield, allowing access to symmetrical bis(alkenyl)stannanes. PMID- 12735745 TI - Novel synthesis of 1-aroyl-3-aryl-4-substituted imidazole-2-thiones. AB - [reaction: see text] An efficient route to 1-aroyl-3-aryl-4-substituted imidazole 2-thiones (2, 4-6) has been developed. The methodology involves the cyclization of 1-aroyl-3-arylthioureas with a variety of carbonyl compounds bearing alpha-H in the presence of bromine and triethylamine. PMID- 12735746 TI - Trimethylsilyl halide-promoted Michaelis-Arbuzov rearrangement. AB - [reaction: see text] We describe a new, straightforward, and easy-to-handle method for achieving an unprecedented trimethylsilyl halide-catalyzed Michaelis Arbuzov-like rearrangement. This rearrangement occurs at temperatures from room temperature to 80 degrees C and does not require addition of any alkyl halide. The scope and limitations of this new reaction are explored, as well as its mechanism. PMID- 12735747 TI - Synthesis of a novel conformationally locked carbocyclic nucleoside ring system. AB - [reaction: see text] A fast and efficient synthetic route to novel Northern locked carbocyclic nucleosides (as precursors of carbocyclic locked nucleic acids or cLNAs) is described. The target nucleoside with a oxabicyclo[2.2.1]heptane ring system was prepared from a simple starting material, diethyl malonate. Ring closure by intramolecular O-alkylation provided the target ring system as the major isomer over the [3.2.0] oxetane system. The adenine moiety was introduced through a reactive triflate after inversion of the stereochemistry of the corresponding alcohol by oxidation and reduction. PMID- 12735749 TI - Bridgehead enolates: substitution and asymmetric desymmetrization of small bridged carbonyl compounds by lithium amide bases. AB - [reaction: see text] Contrary to expectations, a number of bridged carbonyl compounds undergo facile bridgehead metalation with lithium amide bases. Diketone, lactone, lactam, and imide functions are all demonstrated to participate in this type of "bridgehead enolate" chemistry, leading to a range of substituted products. Meso compounds can also be desymmetrized in very high ee by asymmetric bridgehead metalation. PMID- 12735748 TI - Synthesis of soluble donor-acceptor double-cable polymers based on polythiophene and tetracyanoanthraquinodimethane (TCAQ). AB - [structure: see text] Novel suitably functionalized tetracyanoanthraquinodimethane (TCAQ) derivatives covalently linked to thiophene moieties have been synthesized. The thiophene-based monomers have been chemically polymerized and copolymerized to yield new and soluble donor-acceptor double cable polymers. The absorption and emission data reveal that the optical properties can be finely tuned by modifying the ratio of monomers in the copolymerization process. PMID- 12735750 TI - Self-assembly and regioselective photodimerization of anthracene having a dendritic substituent. AB - [reaction: see text] Thermotropic liquid crystallinity can be induced for an anthracene having a dendritic substituent. The photodimerization reaction of an anthracene moiety in the smectic phase proceeded quantitatively and regioselectively. PMID- 12735751 TI - Convergent cyclopentannelation process. AB - [reaction: see text] A triply convergent synthesis of alpha-methylene cyclopentenones has been demonstrated in which an intermediate C,O-dianion is formed and trapped on carbon by electrophiles. PMID- 12735752 TI - Organocatalytic asymmetric assembly reactions: one-pot synthesis of functionalized beta-amino alcohols from aldehydes, ketones, and azodicarboxylates. AB - [reaction: see text] l-Proline catalyzed the enzyme-like direct asymmetric assembly of aldehydes, ketones, and azodicarboxylic acid esters to provide optically active beta-amino alcohols. This assembly reaction uses both aldehydes and ketones as donors in one pot. The aldol-derived stereocenter is formed with a reduced facial selectivity in reactions involving (R)-amino aldehydes. The reactions can be performed on a multigram scale under operationally simple and safe conditions without the requirement of an inert atmosphere or dry solvents. PMID- 12735753 TI - Hetero [6+3] cycloaddition of fulvenes with N-alkylidene glycine esters: a facile synthesis of the delavayine and incarvillateine framework. AB - [reaction: see text] In contrast to the [3+2] or [4+3] cycloaddition of N metalated azomethine ylides and various alkenes, N-benzylidene glycine ethyl ester reacts with fulvenes to give the hetero [6+3] cycloaddition adducts with high stereoselectivity, constituting an efficient and novel route to [2]pyrindines. PMID- 12735754 TI - Enantio- and diastereoselective synthesis of cyclic beta-hydroxy allylsilanes via sequential aldehyde gamma-silylallylboration and ring-closing metathesis reactions. AB - [reaction: see text] Highly enantioenriched cyclic allylsilanes are prepared via stereoselective gamma-silylallylboration reactions of beta- or gamma-unsaturated aldehydes followed by ring-closing metathesis. PMID- 12735755 TI - Common-intermediate strategy for synthesis of conduritols and inositols via beta hydroxy cyclohexenylsilanes. AB - [reaction: see text] Syntheses of conduritols B-D and F and d-(+)-chiro- and neo inositols from cyclohexenylsilane intermediates are described. The key cyclohexylsilane intermediates 5 and 14 were synthesized by stereoselective olefin dihydroxylation of the corresponding cyclohexenylsilanes. Selective Peterson elimination reactions and Fleming-Tamao oxidations of 5 and 14 then delivered the targeted cyclitol derivatives. PMID- 12735756 TI - Intramolecular Diels-Alder reactions of 5-vinyl-1,3-cyclohexadienes. AB - [reaction: see text] The intramolecular Diels-Alder reaction of 5-vinyl-1,3 cyclohexadienes affording substituted tricyclo[3.2.1.0(2,7)]oct-3-enes is studied. PMID- 12735757 TI - Unprecedented palladium-catalyzed cross-coupling reaction of alpha-bromo sulfoxides with boronic acids. AB - [reaction: see text] A new Suzuki-type palladium-catalyzed reaction of boronic acids with alpha-bromo sulfoxides has been developed using a protocol similar to the well-documented reaction of boronic acids with aryl halides. Both cross coupling and homocoupling processes were observed. The best yields in cross coupling products were obtained when the presence of oxygen was carefully excluded using degassed solvents. The oxidative addition palladium complex intermediate could be isolated and characterized by X-ray single-crystal diffraction. PMID- 12735758 TI - Titanium-catalyzed stereoselective geminal heterodihalogenation of beta ketoesters. AB - [reaction: see text] beta-Ketoesters can be effectively monofluorinated with F TEDA using CpTiCl(3) as a catalyst. With the use of this catalyst, the extent of the competing difluorination does not reach 10%. [TiCl(2)(TADDOLato)] complexes catalyze the one-pot enantioselective heterodihalogenation of beta-ketoesters with F-TEDA and NCS to afford alpha-chloro-alpha-fluoro-beta-ketoesters in moderate to good yields. The sequence of addition of the halogenating agents determines the sense of chiral induction. PMID- 12735759 TI - High enantioselectivity in rhodium-catalyzed allylic alkylation of 1-substituted 2-propenyl acetates. AB - [reaction: see text] Rhodium-catalyzed asymmetric allylic alkylation of 1 substituted 2-propenyl acetates with dimethyl malonate proceeded with high enantioselectivity in the presence of cesium carbonate as a base and a rhodium catalyst generated from Rh(dpm)(C(2)H(4))(2) (dpm = dipivaloylmethanato) and a chiral phosphino-oxazoline whose basic skeleton is axially chiral binaphthyl to give branch alkylation products in greater than 90% ee. PMID- 12735760 TI - Palladium-catalyzed synthesis of novel optically active tryptophan analogues. AB - [reaction: see text] Both unsaturated proline derivatives and optically active tryptophan analogues have been obtained via Pd-catalyzed cyclization of aniline containing acetylenic amino acids. The side chain length of the cyclization precursor determines which one of the two possible products will be formed. PMID- 12735761 TI - Synthesis of some biologically relevant beta-C-glycoconjugates. AB - [reaction: see text] An esterification-RCM approach to a variety of biologically relevant beta-C-glycoconjugates is reported herein. A range of carboxylic acids were coupled with several different olefin alcohols 1 to provide esters 3. The esters were then converted to the final ring-closed product 6 in three steps in 49-60% overall yield. The formed compounds are biologically relevant and serve as stable carbohydrate mimics of the corresponding O-glycosides. PMID- 12735762 TI - W(CO)5(L)-catalyzed endo-selective cyclization of allenyl silyl enol ethers: an efficient method for the cyclopentene annulation onto alpha,beta-unsaturated ketones. AB - [reaction: see text] Indium-mediated allenylation of alpha,beta-unsaturated ketones in the presence of tert-butyldimethylsilyl triflate and dimethyl sulfide gives 6-siloxy-1,2,5-trienes, which undergo W(CO)(5)(L)-catalyzed 5-endo cyclization to give the corresponding cyclopentene derivatives in good yield. Furthermore, this novel W(CO)(5)(L)-catalyzed cyclization of allenyl silyl enol ethers proceeds in a 6-endo manner when 5-siloxy-1,2,5-trienes are employed as a substrate. In these reactions, effective electrophilic activation of allenyl compounds for attack by silyl enol ethers is achieved using a catalytic amount of W(CO)(6). PMID- 12735763 TI - Total synthesis of (-)- and (+)-membrenone C. AB - [reaction: see text] A synthesis of the polypropionate marine defense substance (+)-membrenone C and its enantiomer that starts from (S)-2-methyl-3-(tert butyldimethylsilyloxy)propanal is described. Key steps include (1) additions of chiral allenylmetal reagents to effect both chain homologation and the concomitant introduction of four stereo centers, (2) a bis-intramolecular hydrosilylation-oxidation sequence to install beta-hydroxy ketone subunits, and (3) a bis-intramolecular aldol reaction to construct the two dihydropyrone termini. PMID- 12735764 TI - Daphniglaucins A and B, novel polycyclic quaternary alkaloids from Daphniphyllum glaucescens. AB - [structure: see text] Two cytotoxic quaternary Daphniphyllum alkaloids with an unprecedented fused-polycyclic skeleton containing a 1 azoniatetracyclo[5.2.2.0.(1,6)0.(4,9)]undecane ring system, daphniglaucins A (1) and B (2), have been isolated from the leaves of Daphniphyllum glaucescens. Their structures and relative stereochemistry were elucidated on the basis of spectroscopic data. PMID- 12735765 TI - Catalytic crossed Michael cycloisomerization of thioenoates: total synthesis of (+/-)-ricciocarpin A. AB - [reaction: see text] Thioenoates are found to participate in highly chemoselective catalytic crossed Michael cycloisomerization with appendant aryl ketone and enoate partners to afford cyclopentene and cyclohexene products. This methodology has enabled a concise total synthesis of the potent molluscicide (+/ )-ricciocarpin A. PMID- 12735766 TI - Dichloroketene-chiral olefin-based approach to pyrrolizidines: highly stereocontrolled synthesis of (+)-amphorogynine A. AB - [reaction: see text] A highly stereoselective route to (+)-amphorogynine A, a novel pyrrolizidine recently isolated from the New Caledonian plant Amphorogynine spicata, has been realized. The key step in the approach is a diastereoselective [2 + 2] dichloroketene-chiral enol ether cycloaddition (dr >or= 93:7) to access a dichlorocyclobutanone intermediate, which is converted into the alkaloid natural product via a pyrrolidinone derivative. PMID- 12735767 TI - New silyl ether reagents for the absolute stereochemical determination of secondary alcohols. AB - [reaction: see text] Herein we report a new set of silyl ether reagents for determining the enantiomeric purity and absolute stereochemistry of secondary alcohols. These derivatives are easily synthesized, provide straightforward spectroscopic results, and allow for facile recovery of the original chiral alcohol. PMID- 12735769 TI - Click linker: efficient and high-yielding synthesis of a new family of SPOS resins by 1,3-dipolar cycloaddition. AB - [reaction: see text] Highly efficient methodology for the construction of functionalized resins was presented involving 1,3-dipolar cycloaddition as the key reaction step. On the basis of this concept, the first series of click backbone amide linkers were synthesized and the application of the FIMT resin 3f to the parallel synthesis of putative dopaminergic agents was demonstrated leading to the selective receptor ligands 10i and 10s revealing high affinity to the D4 subtype. PMID- 12735768 TI - Biomimetic synthesis of gramicidin s and analogues by enzymatic cyclization of linear precursors on solid support. AB - [reaction: see text] Gramicidin S is a potent decapeptide antibiotic with high hemolytic activity but is unlikely to provoke microbial resistance. Here we demonstrate that gramicidin thioesterase (GrsB TE) correctly cyclizes immobilized linear decapeptide precursors into head-to-tail products, indicating its suitability for parallel solid-phase synthesis of gramicidin analogues from linear precursors on solid support. This chemoenzymatic method will enable the optimization of the therapeutic index of the natural product to fight microbial resistance. PMID- 12735770 TI - Tandem formation and [2,3] rearrangement of methylene ammonium ylides derived from amines and the Simmons-Smith reagent. AB - [reaction: see text] Zinc-complexed methylene ammonium ylides are formed from tertiary amines and the Simmons-Smith reagent. These stable entities can be activated with n-BuLi to allow reactions typical of ammonium ylides such as [2,3] rearrangements. In the case of oxazolidine 12, ylide formation, activation, and subsequent [2,3] rearrangement was highly efficient and occurred with very high diastereoselectivity. PMID- 12735771 TI - A facile route to polysubstitutedN-benzyl pyroglutamates. AB - [reaction: see text] Base-induced coupling/cyclization reactions of alpha sulfonylacetamide with various ethyl (Z)-2-bromo-2-propenoates have been carried out. The polysubstituted pyroglutamate carbon skeleton, with three contiguous asymmetric centers, was built up in one step. A ring-closure mechanism is proposed for the reactions. PMID- 12735772 TI - Synthesis of carbocyclic and heterocyclic fused quinolines by cascade radical annulations of unsaturated N-aryl thiocarbamates, thioamides, and thioureas. AB - [reaction: see text] Tandem radical cyclizations of suitably substituted N-aryl thiocarbamates, thioamides, and thioureas are induced by exposure to tris(trimethylsilyl)silane (TTMSH) and UV light and provide furoquinolines, isofuroquinolines, cyclopentaquinolines, indoloquinolines, and related ring systems. The intermediacy of an alpha-thioalkylamino radical, which is the synthetic equivalent of an imidoyl radical, is invoked. PMID- 12735773 TI - Diastereoselective cyclization in chiral diarylethene crystals: polymorphism and selectivity. AB - [structure: see text] An optically active photochromic diarylethene, (S)-1-(2 methyl-5-phenyl-3-thienyl)-2-[2-methyl-5-(4-(3-methyl-1-penten-1-yl)phenyl)-3 thienyl]perfluorocyclopentene ((S)-1a), was synthesized. (S)-1a formed two crystalline phases, alpha- and beta-phases. The diarylethene underwent a photochromic reaction in solution and even in the single-crystalline phase. In solution, no diastereoselection was observed. On the other hand, in the beta crystalline phase, only one diastereomer (S,R,R)-1b was produced. No such diastereoselection was observed in the alpha-crystalline phase. PMID- 12735774 TI - Indium-mediated reduction of hydroxylamines to amines. AB - [reaction: see text] A novel and simple procedure for reduction of hydroxylamines to the corresponding amines by means of indium powder in aqueous media is reported. Applicability to one-pot reactions and isoxazolidine N-O bond reduction is also demonstrated. A catalytic version of the process using 2-5% In in the presence of other metals (Zn, Al) has been successfully developed. PMID- 12735775 TI - Activation of hydrogen peroxide through hydrogen-bonding interaction with acidic alcohols: epoxidation of alkenes in phenol. AB - [reaction: see text] Electrophilic activation of hydrogen peroxide can be achieved in acidic alcohol solvents without the need for a metal catalyst. This concept is illustrated by the epoxidation of alkenes with H(2)O(2) employing phenol as a solvent. It is proposed that intermolecular hydrogen bonding between H(2)O(2) and phenol activates H(2)O(2) for oxygen-atom transfer. In this interaction, the role of phenol is purely catalytic. PMID- 12735777 TI - Enantioselective total synthesis and structure revision of spirodihydrobenzofuranlactam 1. Total synthesis of stachybotrylactam. AB - [reaction: see text] The enantioselective total synthesis and structure revision of spirodihydrobenzofuranlactam 1 and of its regioisomer 25 are presented. Optically pure (+)-Wieland-Miescher ketone was utilized to construct the AB bicyclic core in 10 steps. Introduction of the resorcylate D-ring unit was achieved by use of our tert-butyl ester metalation sequence. Subsequent stereoselective spirocyclization to form the C-ring was followed by regioselective ring cyanation and lactam formation to produce the pentacyclic structure 1 and its regioisomer 25. PMID- 12735776 TI - Synthesis of isobutyl-C-galactoside (IBCG) as an isopropylthiogalactoside (IPTG) substitute for increased induction of protein expression. AB - [reaction: see text] Addition of isopropyl-beta-d-thiogalactopyranoside (IPTG) to bacterial cultures is often used to induce expression of plasmid-based genes for the production of recombinant proteins under control of the lac promoter, but a simple method to circumvent the inherent instability of this compound has not been addressed experimentally. Herein we report the first synthesis of isobutyl-C galactoside (IBCG), the C-glycoside analogue of IPTG, and show that IBCG is superior to IPTG in inducing protein expression over long induction times. PMID- 12735778 TI - The C2 selective nucleophilic substitution reactions of 2,3-epoxy alcohols mediated by trialkyl borates: the first endo-mode epoxide-opening reaction through an intramolecular metal chelate. AB - [reaction: see text] Highly efficient C2 selective substitution reactions of 2,3 epoxy alcohols with nucleophiles were developed by using NaN(3)-(CH(3)O)(3)B, NaSPh-(CH(3)O)(3)B, or NaCN-(C(2)H(5)O)(3)B system. The reaction proceeds through novel endo-mode epoxide opening of an intramolecular boron chelate, which was suggested from both experimental and quantum mechanic studies. PMID- 12735779 TI - Intermolecular enol ether-alkyne metathesis. AB - [reaction: see text] A new intermolecular metathesis between a variety of alkynes and alkyl enol ethers is described. The best results were obtained with the second generation Grubbs' precatalysts. The reaction occurred with a high degree of regioselectivity and provided electron-rich dienes, which underwent a variety of [4 + 2] cycloadditions. PMID- 12735780 TI - Initiator-dependent chemoselective addition of THF radical to aldehyde and aldimine and its application to a three-component reaction. AB - [reaction: see text] The product distribution of the three-component reaction of aldehydes, arylamines, and THF was dependent on a radical initiator, preferentially giving the corresponding THF adducts of imines with dimethylzinc and adducts of aldehyde with triethylborane. PMID- 12735781 TI - Total synthesis of debromoflustramine B via biomimetic alkylative cyclization. AB - [reaction: see text] The hexahydropyrrolo[2,3-b]indoline skeleton is readily accessed by the zinc triflate-mediated alkylation of tryptamine derivatives. The methodology was employed in a three-step total synthesis of debromoflustramine B. PMID- 12735782 TI - Tandem Diels-Alder/fragmentation approach to the synthesis of eleutherobin. AB - [reaction: see text] A synthesis of 28, the carbon framework of the eleutherobin aglycone, is reported in a 15-step sequence from readily available starting materials. The tandem Diels-Alder reaction of 6 and 7 to produce 18, in which three new rings and six new stereocenters are formed, is a key step in the reaction sequence. PMID- 12735783 TI - Medication errors caused by confusion of drug names. AB - Many drug names can look or sound like other drug names, which leads to confusion and potentially harmful medication errors. While various types of drug names exist, brand (proprietary) names are most commonly confused. Examples of the numerous drug names that have been confused because they look and/or sound similar include Celebrex (celecoxib), Cerebyx (fosphenytoin), and Celexa (citalopram). Factors such as poor handwriting and clinical similarity may exacerbate the problem. This problem can be alleviated through actions by regulatory agencies, pharmaceutical manufacturers, healthcare professionals, and patients. To address the problem, significant changes in the pharmaceutical regulatory process have occurred in the US and Europe. PMID- 12735784 TI - Cardiovascular safety of sildenafil. AB - Initial reports of myocardial infarction and sudden death in men with erectile dysfunction who had taken sildenafil (sometimes in conjunction with nitrates) raised concerns that sildenafil may increase the risk of cardiovascular events in men with erectile dysfunction and vascular disease. A significant body of evidence now indicates that sildenafil generally has a good safety profile in men with erectile dysfunction and cardiovascular disease. Sildenafil therapy does not appear to be associated with ischaemic events either at the time of introduction of therapy or during longer-term use. Rates of discontinuation from sildenafil therapy due to adverse events are similar to placebo in men with cardiovascular disease. Sildenafil does not interact in a potentially hazardous way with antihypertensive or antianginal therapy, with the exception of nitrates. Nitrates should not be administered within 24 hours of sildenafil therapy, and care should be taken to determine whether sildenafil may have been used before nitrates are administered to patients. Sildenafil appears to be generally well tolerated in most patients with chronic, stable cardiovascular disease. PMID- 12735787 TI - Important drug safety information on the internet: assessing its accuracy and reliability. AB - BACKGROUND: The Internet is becoming increasingly important as a source of health related information, but the accuracy and reliability of information presented on the world wide web is debated. OBJECTIVE: We aimed to assess whether important, recent drug safety information is accurately reflected on Internet sites. METHODS: We evaluated whether major warnings issued by the US FDA between October 1, 2000 and September 30, 2001 on severe and life-threatening drug toxicity were mentioned 4-16 months later in the top ten web pages identified for these drugs by each of seven different search engines. We examined predictors of precise mention of the FDA warnings using logistic regressions. RESULTS: Twenty major safety warnings on 21 drugs (including three withdrawals) were eligible for the study. Among 519 different pertinent web pages retrieved (16-32 for each drug), precise mention of the safety issue was made in only 165 (31.8%). Best rates of precise mention were seen in web sites sponsored by attorneys (79.4%), in physician-oriented web pages (65.5%) and for withdrawn drugs (57.9%). In addition to these factors, better coverage of the FDA warnings was independently seen when no other adverse effects from the same organ system was mentioned (p < 0.001), while coverage was worse when there was no date on the site and web page (p = 0.020), and when the site owner could not be classified or was unknown (p = 0.014). CONCLUSIONS: Important safety warnings are inadequately covered in the majority of web pages. This deficiency creates a source of potentially harmful misinformation for health consumers. PMID- 12735789 TI - Effects of a comprehensive cardiac rehabilitation program on quality of life and exercise tolerance in women: A retrospective analysis. AB - BACKGROUND: Currently, there are a lack of investigations that have examined the effect of participating in a comprehensive cardiac rehabilitation program on quality of life and physiological measures in women of different ages. The purpose of this investigation was to examine the effect of participating in a comprehensive cardiac rehabilitation program on quality of life, exercise tolerance, blood pressure and lipids in women between 33 and 82 years of age. METHODS: The 126 women participated in a 14-week cardiac rehabilitation program that consisted of 7 weeks of formal supervised exercise training and 7 weeks of unsupervised exercise and lifestyle modification. Physiologic and quality of life outcome measures obtained at the outset and after 14 weeks included: 1) exercise treadmill time; 2) resting and peak systolic and diastolic blood pressure; 3) total cholesterol, low-density lipoprotein cholesterol, high density lipoprotein cholesterol and Triglycerides; 4) Cardiac Quality of Life Index questionnaire. RESULTS: Significant improvements were found in the following quality of life measures after participating in the cardiac rehabilitation program: physical well being, psychosocial, worry, nutrition and symptoms. No significant differences were seen for any QOL variable between the different age groups. Significant improvements were seen in exercise tolerance (+21%) and high density lipoprotein (+5%). CONCLUSION: Cardiac rehabilitation may play an important role in improving quality of life, exercise tolerance and high density lipoprotein cholesterol levels in younger and older women with underlying cardiovascular disease. PMID- 12735785 TI - Benefits and risks of pharmacological treatments for essential tremor. AB - Essential tremor can cause significant functional disability in some patients. The arms are the most common body part affected and cause the most functional disability. The treatment of essential tremor includes medications, surgical options and other forms of therapy. Presently there is no cure for essential tremor nor are there any medications that can slow the progression of tremor. Treatment for essential tremor is recommended if the tremor causes functional disability. If the tremor is disabling only during periods of stress and anxiety, propranolol and benzodiazepines can be used during those periods when the tremor causes functional disability. The currently available medications can improve tremor in approximately 50% of the patients. If the tremor is disabling, treatment should be initiated with either primidone or propranolol. If either primidone or propranolol do not provide adequate control of the tremor, then the medications can be used in combination. If patients experience adverse effects with propranolol, occasionally other beta-adrenoceptor antagonists (such as atenolol or metoprolol) can be used. If primidone and propranolol do not provide adequate control of tremor, occasionally the use of benzodiazepines (such as clonazepam) can provide benefit. Other medications that may be helpful include gabapentin or topiramate. If a patient has disabling head or voice tremor, botulinum toxin injections into the muscles may provide relief from the tremor. Botulinum toxin in the hand muscles for hand tremor can result in bothersome hand weakness and is not widely used. There are other medications that have been tried in essential tremor and have questionable efficacy. These drugs include carbonic anhydrase inhibitors (e.g. methazolamide), phenobarbital, calcium channel antagonists (e.g. nimodipine), isoniazid, clonidine, clozapine and mirtazapine. If the patient still has disabling tremor after medication trials, surgical options are usually considered. Surgical options include thalamotomy and deep brain stimulation of the thalamus. These surgical options provide adequate tremor control in approximately 90% of the patients. Surgical morbidity and mortality for these procedures is low. Deep brain stimulation and thalamotomy have been shown to have comparable efficacy but fewer complications have been reported with deep brain stimulation. In patients undergoing bilateral procedures deep brain stimulation of the thalamus is the procedure of choice to avoid adverse effects seen with bilateral ablative procedures. The use of medication and/or surgery can provide adequate tremor control in the majority of the patients. PMID- 12735790 TI - A markup language for electrocardiogram data acquisition and analysis (ecgML). AB - BACKGROUND: The storage and distribution of electrocardiogram data is based on different formats. There is a need to promote the development of standards for their exchange and analysis. Such models should be platform-/ system- and application-independent, flexible and open to every member of the scientific community. METHODS: A minimum set of information for the representation and storage of electrocardiogram signals has been synthesised from existing recommendations. This specification is encoded into an XML-vocabulary. The model may aid in a flexible exchange and analysis of electrocardiogram information. RESULTS: Based on advantages of XML technologies, ecgML has the ability to present a system-, application- and format-independent solution for representation and exchange of electrocardiogram data. The distinction between the proposal developed by the U.S Food and Drug Administration and ecgML model is given. A series of tools, which aim to facilitate ecgML-based applications, are presented. CONCLUSIONS: The models proposed here can facilitate the generation of a data format, which opens ways for better and clearer interpretation by both humans and machines. Its structured and transparent organisation will allow researchers to expand and test its capabilities in different application domains. The specification and programs for this protocol are publicly available. PMID- 12735788 TI - Small glutamine-rich tetratricopeptide repeat-containing protein (SGT) interacts with the ubiquitin-dependent endocytosis (UbE) motif of the growth hormone receptor. AB - Endocytosis of the growth hormone receptor (GHR) is regulated by the ubiquitin conjugating system. A cytosolic 10 amino acid motif, referred to as the ubiquitin dependent endocytosis (UbE) motif, is involved in the ubiquitination as well as in the endocytosis of the receptor. Proteins that are implicated in one of these processes have not been identified so far. Using a glutathione S-transferase (GST)-pulldown assay with a GST fusion protein encompassing the UbE motif of the GHR, a 35 kDa protein was purified. The protein was identified by MS as small glutamine-rich tetratricopeptide repeat (TPR)-containing protein (SGT). We found that GHR interacts with SGT. In vivo, both the precursor and the mature form of the receptor interacted with SGT. Inactivation of the ubiquitin-conjugating system did not affect the GHR-SGT interaction. Binding studies showed that the first TPR motif of SGT interacts with the UbE motif of the GHR. Taken together, these data show that SGT is a GHR-interacting protein, which binds independent of the ubiquitin-conjugating system. PMID- 12735786 TI - Benefit-risk assessment of antileukotrienes in the management of asthma. AB - Antileukotrienes are a relatively new class of anti-asthma drugs that either block leukotriene synthesis (5-lipoxygenase inhibitors) like zileuton, or antagonise the most relevant of their receptors (the cysteinyl leukotriene 1 receptor [CysLT1]) like montelukast, zafirlukast or pranlukast. Hence, their major effect is an anti-inflammatory one. With the exception of pranlukast, the other antileukotrienes have been studied and marketed in the US and Europe for long enough to establish that they are useful drugs in the management of asthma. Their effects, significantly better than placebo, seem more pronounced in subjective measurements (i.e. symptoms scores or quality-of-life tests) than in objective parameters (i.e. forced expiratory volume in 1 second or peak expiratory flow rate). Also, there is some evidence that these drugs work better in some subsets of patients with certain genetic polymorphisms - probably related to their leukotriene metabolism - or patients with certain asthma characteristics. There are a small number of comparative studies only, and with regard to long-term asthma control differences between the agents have not been evaluated. Nevertheless, their overall effect appears comparable with sodium cromoglycate (cromolyn sodium) or theophylline, but significantly less than low dose inhaled corticosteroids. Antileukotrienes have been shown to have a degree of corticosteroid-sparing effect, but salmeterol appears to perform better as an add-on drug. Montelukast is probably the most useful antileukotriene for continuous treatment of exercise-induced asthma, performing as well as salmeterol without inducing any tolerance. All antileukotrienes are taken orally; their frequency of administration is quite different ranging from four times daily (zileuton) to once daily (montelukast). Antileukotrienes are well tolerated drugs, even though zileuton intake has been related to transitional liver enzyme elevations in some cases. Also Churg-Strauss syndrome (a systemic vasculitis), has been described in small numbers of patients taking CysLT1 antagonists. It is quite probable that this disease appears as a consequence of an 'unmasking' effect when corticosteroid dosages are reduced in patients with severe asthma once CysLT1 antagonists are introduced, but more data are needed to definitely establish the mechanism behind this effect. Overall, however, the benefits of antileukotrienes in the treatment of asthma greatly outweigh their risks. PMID- 12735791 TI - Oral health knowledge and behavior among male health sciences college students in Kuwait. AB - BACKGROUND: Health auxiliary personnel have an important role in oral health promotion when they graduate and start working in the health care system. This study aims to find out oral health knowledge and oral health behavior of male Health Sciences College students. METHODS: A questionnaire was distributed to all students at the male Health Sciences College in Kuwait (N = 153) during the academic year 2001/2002. The students filled the anonymous questionnaire in the class after the lecture. The response rate was 84% (n = 128). The questions consisted information on the general background, oral health behavior and oral health knowledge. RESULTS: Oral health knowledge seemed to be limited and very few background factors were associated with it. More than half of the students had visited a dentist during the previous 12 months, but only one third of students were brushing twice a day or more often. CONCLUSIONS: It may be concluded that the male Health Sciences College students seemed to have appropriate knowledge on some oral health topics, but limited knowledge on the others. Their toothbrushing practices are still far behind the international recommendation (twice a day) and also the knowledge, why it should be done so frequently also very limited. PMID- 12735792 TI - Improved immunohistochemical evaluation of micrometastases in sentinel lymph nodes of cutaneous melanoma with 'MCW melanoma cocktail'--a mixture of monoclonal antibodies to MART-1, Melan-A, and tyrosinase. AB - BACKGROUND: MART-1, Melan-A, and Tyrosinase have shown encouraging results for evaluation of melanoma micrometastases in sentinel lymph nodes, as compared to conventionally used S-100 protein and HMB-45. To achieve higher sensitivity, some studies recommend evaluation of three sections, each at intervals of 200 micron. This would mean, routine staining of three adjacent sections in each of the three clusters at intervals of 200 micron, requiring nine slides resulting in added expense. If a cocktail of these antibodies could be used, only one section would be required instead of three generating significant cost savings. METHODS: We prepared a combination of monoclonal antibodies to these three immunomarkers in optimized dilutions (MART-1, clone M2-7C10, dilution 1:500; Melan-A, clone A103, dilution 1:100; and Tyrosinase, clone T311, dilution 1:50) and designated it as 'MCW melanoma cocktail'. Formalin-fixed paraffin-embedded tissue sections of sentinel lymph nodes from patients with cutaneous melanoma, without macro metastases were evaluated with this cocktail. RESULTS: Melanoma micrometastases were easily detectable with the cocktail in 41 out of 188 slices (8/24 cases). The diagnostic accuracy amongst five pathologists did not show statistically significant difference. Out of 188 slices, 78 had adjacent sections immunostained individually with MART-1 and Melan-A during our previous study. Of these 78 slices, 21 were positive for melanoma micrometastases with MART-1 and Melan-A individually. However, the adjacent section of these slices immunostained with the cocktail detected metastases in four additional slices. Thus, MART-1 and Melan-A could not detect melanoma micrometastases individually in 16% (4/25) of slices positive with the cocktail. Benign capsular nevi were immunoreactive for the cocktail in 4.8% (9/188) slices. All 81 slices of negative test controls (sentinel lymph nodes of mammary carcinoma) were interpreted correctly as negative for melanoma micrometastases. CONCLUSIONS: The melanoma cocktail facilitated easy interpretation of melanoma micrometastases in sentinel lymph nodes with high interobserver agreement. There was improvement in detection rate with the cocktail as compared to MART-1 and Melan-A individually. Furthermore, this approach facilitates cost savings. PMID- 12735793 TI - A comparison of Child-Pugh, APACHE II and APACHE III scoring systems in predicting hospital mortality of patients with liver cirrhosis. AB - BACKGROUND: The aim of this study was to assess the prognostic accuracy of Child Pugh and APACHE II and III scoring systems in predicting short-term, hospital mortality of patients with liver cirrhosis. METHODS: 200 admissions of 147 cirrhotic patients (44% viral-associated liver cirrhosis, 33% alcoholic, 18.5% cryptogenic, 4.5% both viral and alcoholic) were studied prospectively. Clinical and laboratory data conforming to the Child-Pugh, APACHE II and III scores were recorded on day 1 for all patients. Discrimination was evaluated using receiver operating characteristic (ROC) curves and area under a ROC curve (AUC). Calibration was estimated using the Hosmer-Lemeshow goodness-of-fit test. RESULTS: Overall mortality was 11.5%. The mean Child-Pugh, APACHE II and III scores for survivors were found to be significantly lower than those of nonsurvivors. Discrimination was excellent for Child-Pugh (ROC AUC: 0.859) and APACHE III (ROC AUC: 0.816) scores, and acceptable for APACHE II score (ROC AUC: 0.759). Although the Hosmer-Lemeshow statistic revealed adequate goodness-of-fit for Child-Pugh score (P = 0.192), this was not the case for APACHE II and III scores (P = 0.004 and 0.003 respectively) CONCLUSION: Our results indicate that, of the three models, Child-Pugh score had the least statistically significant discrepancy between predicted and observed mortality across the strata of increasing predicting mortality. This supports the hypothesis that APACHE scores do not work accurately outside ICU settings. PMID- 12735794 TI - Presence and expression of hydrogenase specific C-terminal endopeptidases in cyanobacteria. AB - BACKGROUND: Hydrogenases catalyze the simplest of all chemical reactions: the reduction of protons to molecular hydrogen or vice versa. Cyanobacteria can express an uptake, a bidirectional or both NiFe-hydrogenases. Maturation of those depends on accessory proteins encoded by hyp-genes. The last maturation step involves the cleavage of a ca. 30 amino acid long peptide from the large subunit by a C-terminal endopeptidase. Until know, nothing is known about the maturation of cyanobacterial NiFe-hydrogenases. The availability of three complete cyanobacterial genome sequences from strains with either only the uptake (Nostoc punctiforme ATCC 29133/PCC 73102), only the bidirectional (Synechocystis PCC 6803) or both NiFe-hydrogenases (Anabaena PCC 7120) prompted us to mine these genomes for hydrogenase maturation related genes. In this communication we focus on the presence and the expression of the NiFe-hydrogenases and the corresponding C-terminal endopeptidases, in the three strains mentioned above. RESULTS: We identified genes encoding putative cyanobacterial hydrogenase specific C-terminal endopeptidases in all analyzed cyanobacterial genomes. The genes are not part of any known hydrogenase related gene cluster. The derived amino acid sequences show only low similarity (28-41%) to the well-analyzed hydrogenase specific C-terminal endopeptidase HybD from Escherichia coli, the crystal structure of which is known. However, computational secondary and tertiary structure modeling revealed the presence of conserved structural patterns around the highly conserved active site. Gene expression analysis shows that the endopeptidase encoding genes are expressed under both nitrogen-fixing and non-nitrogen-fixing conditions. CONCLUSION: Anabaena PCC 7120 possesses two NiFe-hydrogenases and two hydrogenase specific C-terminal endopeptidases but only one set of hyp-genes. Thus, in contrast to the Hyp-proteins, the C-terminal endopeptidases are the only known hydrogenase maturation factors that are specific. Therefore, in accordance with previous nomenclature, we propose the gene names hoxW and hupW for the bidirectional and uptake hydrogenase processing endopeptidases, respectively. Due to their constitutive expression we expect that, at least in cyanobacteria, the endopeptidases take over multiple functions. PMID- 12735796 TI - Gene structure and chromosomal localization of mouse Opa1 : its exclusion from the Bst locus. AB - BACKGROUND: Autosomal dominant optic atrophy type 1 (DOA) is the most common form of hereditary optic atrophy in human. We have previously identified the OPA1 gene and shown that it was mutated in patients with DOA. OPA1 is a novel member of the dynamin GTPase family that play a role in the distribution of the mitochondrial network. The Bst (belly spot and tail) mutant mice show atrophy of the optic nerves and previous mapping data raise the possibility that Bst and OPA1 are orthologs. In order to analyse the Bst mouse as a model for DOA, we therefore characterized mouse Opa1 and evaluated it as a candidate for the Bst mutant mouse. RESULTS: Comparison of mouse and human OPA1 sequences revealed 88% and 97% identity at the nucleotide and amino acid levels, respectively. Presence of alternatively spliced mRNAs as seen in human was conserved in the mouse. Screening of the whole mRNA coding sequence and of the 31 exons of Opa1 did not reveal any mutation in Bst. Using a radiation hybrid panel (T31), we mapped Opa1 to chromosome 16 between genetic markers D16Mit3 and D16Mit124, which is 10 cM centromeric to the Bst locus. CONCLUSION: On the basis of these results we conclude that Opa1 and Bst are distinct genes and that the Bst mouse is not the mouse model for DOA. PMID- 12735799 TI - Value of repeat ultrasonic prostatic biopsies following negative digitally directed biopsy - page 289. PMID- 12735797 TI - Molecular evolution of adenylating domain of aminoadipate reductase. AB - BACKGROUND: Aminoadipate reductase (Lys2) is a fungal-specific protein. This enzyme contains an adenylating domain. A similar primary structure can be found in some bacterial antibiotic/peptide synthetases. In this study, we aimed to determine which bacterial adenylating domain is most closely related to Lys2. In addition, we analyzed the substitution rate of the adenylating domain-encoding region. RESULTS: Some bacterial proteins contain more than two similar sequences to that of the adenylating domain of Lys2. We compared 67 amino acid sequences from 37 bacterial and 10 fungal proteins. Phylogenetic trees revealed that the lys2 genes are monophyletic; on the other hand, bacterial antibiotic/peptide synthase genes were not found to be monophyletic. Comparative phylogenetic studies among closely related fungal lys2 genes showed that the rate of insertion/deletion in these genes was lower and the nucleotide substitution rate was higher than that in the internal transcribed spacer (ITS) regions. CONCLUSIONS: The lys2 gene is one of the most useful tools for revealing the phylogenetic relationships among fungi, due to its low insertion/deletion rate and its high substitution rate. Lys2 is most closely related to certain bacterial antibiotic/peptide synthetases, but a common ancestor of Lys2 and these synthetases evolutionarily branched off in the distant past. PMID- 12735798 TI - The yeast ISN1 (YOR155c) gene encodes a new type of IMP-specific 5'-nucleotidase. AB - BACKGROUND: The purine salvage enzyme inosine 5'-monophosphate (IMP)-specific 5' nucleotidase catalyzes degradation of IMP to inosine. Although this enzymatic activity has been purified and characterized in Saccharomyces cerevisiae, the gene encoding IMP 5'-nucleotidase had not been identified. RESULTS: Mass spectrometry analysis of several peptides of this enzyme purified from yeast allowed identification of the corresponding gene as YOR155c, an open reading frame of unknown function, renamed ISN1. The deduced Isn1p sequence was clearly not homologous to 5'-nucleotidases from other species. However, significant similarities to Isn1p were found in proteins of unknown function from Neurospora crassa, Plasmodium falciparum and several yeast species. Knock-out of ISN1 resulted in the total loss of IMP-specific 5'-nucleotidase activity, thus confirming that the ISN1 gene indeed encodes the enzymatic activity purified from yeast. In vivo studies revealed that, when IMP is overproduced through constitutive activation of the IMP de novo synthesis pathway, ISN1 is required for excretion of inosine and hypoxanthine in the medium. CONCLUSION: We have identified a new yeast gene, ISN1 (YOR155c), as encoding IMP-specific 5' nucleotidase activity. The ISN1 gene defines a new type of 5'-nucleotidase which was demonstrated to be functional in vivo. PMID- 12735800 TI - Meta-analysis of four different surgical treatments for stress urinary incontinence - Page 300. PMID- 12735795 TI - The gene structure and expression of human ABHD1: overlapping polyadenylation signal sequence with Sec12. AB - BACKGROUND: Overlapping sense/antisense genes orientated in a tail-to-tail manner, often involving only the 3'UTRs, form the majority of gene pairs in mammalian genomes and can lead to the formation of double-stranded RNA that triggers the destruction of homologous mRNAs. Overlapping polyadenylation signal sequences have not been described previously. RESULTS: An instance of gene overlap has been found involving a shared single functional polyadenylation site. The genes involved are the human alpha/beta hydrolase domain containing gene 1 (ABHD1) and Sec12 genes. The nine exon human ABHD1 gene is located on chromosome 2p23.3 and encodes a 405-residue protein containing a catalytic triad analogous to that present in serine proteases. The Sec12 protein promotes efficient guanine nucleotide exchange on the Sar1 GTPase in the ER. Their sequences overlap for 42 bp in the 3'UTR in an antisense manner. Analysis by 3' RACE identified a single functional polyadenylation site, ATTAAA, within the 3'UTR of ABHD1 and a single polyadenylation signal, AATAAA, within the 3'UTR of Sec12. These polyadenylation signals overlap, sharing three bp. They are also conserved in mouse and rat. ABHD1 was expressed in all tissues and cells examined, but levels of ABHD1 varied greatly, being high in skeletal muscle and testis and low in spleen and fibroblasts. CONCLUSIONS: Mammalian ABHD1 and Sec12 genes contain a conserved 42 bp overlap in their 3'UTR, and share a conserved TTTATTAAA/TTTAATAAA sequence that serves as a polyadenylation signal for both genes. No inverse correlation between the respective levels of ABHD1 and Sec12 RNA was found to indicate that any RNA interference occurred. PMID- 12735801 TI - Value of repeat ultrasonic prostatic biopsies following negative digitally directed biopsy. AB - OBJECTIVE: The objective of this study was to determine the benefit of repeat transrectal ultrasound-guided prostatic biopsy (TRUSBx) among men with prior benign histology on digitally guided biopsy despite suspicious digital rectal examination (DRE) findings. PATIENTS AND METHODS: From January 1, 1990 to May 30, 1993, we evaluated 130 consecutive men, referred to us with benign pathology on digitally-guided biopsy and DRE suspicious of cancer. All patients underwent systematic and directed TRUSBx. RESULTS: TRUSBx detected previously undiagnosed malignancy in 67 cases (51%). CONCLUSION: It is mandatory to reevaluate by TRUSBx, all patients with a palpable abnormality on DRE and a prior benign pathology on digitally-guided biopsy, as the likelihood of finding cancer is elevated. PMID- 12735802 TI - Receiver operating characteristic (ROC) curve analysis of transrectal ultrasonographic prostate cancer detection. AB - PURPOSE: To provide an overview of receiver operating characteristics (ROC) curve theory and create an ROC curve describing transrectal ultrasound (TRUS) detection of prostate cancer (PCa). METHODS: One hundred thirty-two patients with either an abnormal digital rectal exam (DRE) or a prostate-specific antigen (PSA) above 4 ng/ml or both underwent TRUS and biopsy of the peripheral zone (PZ) of each quadrant. ROC software was used to create an ROC curve. RESULTS: One hundred seventy-nine of 528 quadrants (34%) harbored PCa. The performance of TRUS in detection of PCa as defined by the area below the ROC is 0,809. CONCLUSIONS: Future investigators are encouraged to employ ROC analysis of TRUS to permit more meaningful comparisons of performance. PMID- 12735803 TI - Meta-analysis of four different surgical treatments for stress urinary incontinence. AB - INTRODUCTION: Multiple surgical approaches for the correction of stress urinary incontinence (SUI) have been described. In an attempt to determine which procedure may be superior, we performed a meta-analysis comparing the needle suspensions (i.e. Gittes, Stamey), Raz, Burch/MMK colposuspension, and urethral sling. METHODS: The most complete 87 of 310 articles relating to the surgical therapy for SUI published between 1982 and 1995 were selected by Medline search for review. These articles incorporated patients operated on from 1965 to 1995. Fixed effects Baysian and variance weighted models were applied to derive estimates and confidence intervals. Bonferroni multiple comparison procedures wee also incorporated. RESULTS: The mean reported success rates were 81% for the needle suspensions, 84% for the Raz, 85% for the colposuspension, and 85% for the sling. There was a significant difference between the sling and needle suspension as well as between the colposuspension and needle suspension (P<0.05). There was no correlation between length of follow-up and the success rate (P>.05). CONCLUSION: There is currently a general impression among urologists that the sling has the best success rates for the treatment of SUI and that the Raz and colposuspension are clinically superior to the needle suspension. Recent literature also indicates a significant increase in the failure rate over time. Our meta-analysis of the best 25% of SUI papers does not support either view entirely. This may be due to publication bias and incomplete reporting by the authors. Prior to developing ever "improved" procedures, urologist must start to report their results in a consistent fashion to allow meaningful conclusions concerning the risks and benefits of our existing procedures. PMID- 12735804 TI - Evaluation of a non-invasive method to determine bladder volume in children. AB - Although urethral catheterization is the "gold standard" for accurate bladder urine volume measurement in all patients, the discomfort associated with its use makes it less than ideal in pediatric urologic practice. Portable ultrasound scanners have been demonstrated to be an acceptable alternative in adults in several studies, but little data exists on their use in children. A portable ultrasound scanner that automatically calculates the bladder urine volume was used for 40 separate determinations and correlated with bladder urine volume measured by urethral catheterization (coefficient of correlation = 0.759). The scanner is reasonably accurate, well-tolerated and a useful alternative to urethral catheterization for measuring bladder urine volume. PMID- 12735805 TI - Gasless laparoscopy for the urologist. AB - PURPOSE: To explore the use of gasless laparoscopy in urology. MATERIALS AND METHODS: Over 120 patients underwent various urological procedures with this technique from September 1994. For extraperitoneal procedures, balloon dissection was used to develop the extraperitoneal space. A mechanical retraction system was used with conventional laparotomy instruments to perform laparoscopic Burch bladder neck suspensions, staging pelvic lymphadenectomies, two-team abdominovaginal slings, nephrectomies, and ureterectomy. RESULTS: Early results indicate that this gasless technique may result in much quicker operative times with similar success rates to the conventional carbon dioxide laparoscopy with respect to bladder neck suspensions. Conversion to an open procedure was also lower with this technique. CONCLUSIONS: Gasless laparoscopic techniques may facilitate urological procedures, and may be easier and quicker to learn. Previous multiple surgeries and excess weight are not contraindications to performing this laparoscopic technique. PMID- 12735806 TI - EDITORIAL. PMID- 12735807 TI - Drug resistance amongst uropathogens isolated from women in a suburban population: laboratory findings over 7 years. AB - Urinary tract infections (UTI) are a common reason for patient visits to the family physician. The following study was carried out during one month in 1989 and one month in 1996, whereby urine specimens from adult females (living in suburban areas of Toronto, Canada), with presumed signs and symptoms of UTI, were processed. The pool of 20 infecting uropathogenic species was relatively unaltered over the seven years. Most isolates were Gram negative pathogens (72% in 1989; 76% in 1996), with Escherichia coli the most common (56.5% in 1989; 61.9%, in 1996), followed by Enterococcus faecalis (17.4% in 1989; 9.47% in 1996). The resistance to commonly prescribed trimethoprim-sulfamethoxazole increased during the seven years from 10% to 15%, for E. coli (Table 2) and from 35% to 100% for enterococci (Table 3). In 1996, although there was resistance to norfloxacin, the top five uropathogenic species were highly susceptible. No enterococci were resistant to nitrofurantoin and almost all E. coli (99.2%) were susceptible. Based upon this in-vitro data, nitrofurantoin, and not trimethoprim sulfamethoxazole, would be the antimicrobial agent of choice for treatment of uncomplicated UTI in adult females. PMID- 12735808 TI - What the prostate cancer patient should know: variations in urologists' opinions. AB - OBJECTIVES: To determine the questions that treating Ontario urologists think should be addressed with patients who have curable prostate cancer before treatment decisions are made. METHODS: All Ontario (Canada) urologists (179) were given a scenario describing a case of curable prostate cancer and asked to judge the importance, using one of four categories (essential/important/no opinion/avoid), of addressing each of 78 questions. RESULTS: Ninety-seven (54%) of urologists completed the survey. The questions identified most frequently as "essential" to address were: treatment-related incontinence rates (76%), cure rates (74%), treatment-related impotence rates (73%), right of refusal (71%), and mortality from watchful waiting (63%). The questions identified most frequently to "avoid" related to searching for the first medical centre (29%) and the number of prostate cancer patients cured by the physicians (16%). There was little agreement among the urologists on the importance of addressing most of the remaining questions (Average agreement 40.7%). CONCLUSIONS: Most urologists assign great importance to addressing questions related to major treatment morbidity and success rates. Beyond these few major categories, there is widespread disparity among urologists about what information the patient should know. This "information gap" presents an identifiable dilemma in our adherence to the ethical principles underpinning informed consent in patients with potentially curable prostate cancer. PMID- 12735809 TI - Hydronephrosis after simple hysterectomy for benign disease: a prospective study. AB - OBJECTIVES: To determine the prevalence and implications of transient asymptomatic hydronephrosis following simple hysterectomy for benign disease. METHODS: Over a 12-month period, 50 consecutive women undergoing total abdominal hysterectomy with or without bilateral salpingo-oophorectomy were evaluated. Renal ultrasound to detect asymptomatic hydronephrosis, renal function tests and urinalysis were performed preoperatively and at 3 days and 6 weeks after surgery. RESULTS: Hydronephrosis was not identified in the early or late postoperative period in any of the 50 evaluated patients. Concomitant bilateral salpingo oophorectomy did not have any impact on this result. CONCLUSIONS: We did not find any cases of transient asymptomatic hydronephrosis occurring after simple, uncomplicated hysterectomy for benign disease. This suggests that silent ureteral dysfunction does not occur as a result of the surgical technique itself. As such, any postoperative finding of asymptomatic hydronephrosis warrants further investigation. PMID- 12735810 TI - A horseshoe kidney harboring large symptomatic renal angiomyolipoma with a false macro-aneurysm. AB - We report a case of a large angiomyolipoma in a horseshoe kidney associated with a macro-aneurysm bleeding into the retroperitoneum. The patient was successfully treated by embolization of the macro-aneurysm, using thrombogenic cois and Ivalon particles. He remains free of symptoms at 18 months of follow-up. CT scan and renal ultrasound confirm no flow to the previous area of the macro-aneurysm and no change in size of the benign tumor. PMID- 12735811 TI - Urachal adenocarcinoma. AB - Primary adenocarcinoma of the bladder is a rare neoplasm comprising less than 2% of all bladder tumors. Approximately one-third of primary bladder adenocarcinoma arise in the urachal remnant. Differentiation from vesical adenocarcinoma preoperatively is essential because the surgical approach differs. We herein present a case of urachal adenocarcinoma whose radiographic work-up permitted the correct preoperative diagnosis. PMID- 12735812 TI - Hypertension and nephritic urine sediment following retroperitoneal lymph node dissection. AB - Retroperitonela lymph node dissection (RPLND), cisplatin-based chemotherapy, and tumor surveillance has dramatically improved survival of patients with non seminomatous germ cell tumors. Complications, including renal vascular injuries, have been encountered with post-chemotherapy RPLND. We report on a patient with delayed renovascular hypertension and nephritic sediment following RPLND. A thirty year old man presented with well-controlled hypertension following treatment of a left testicular non-seminomatous germ cell tumor 11 years earlier. Post-orchiectomy investigation revealed retroperitoneal lymphadenopathy which was treated with cispaltin-based chemotherapy. A residual mass was managed surgically and during perihilar dissection, the left renal vein was injured and repaired. Current investigation revealed hypertension and a nephritic urine sediment, both of which resolved with left simple nephrectomy. We believe that inadvertent renal artery injury was responsible for this late complication. PMID- 12735813 TI - Genital hair strangulation in an uncircumcised boy. AB - Penile strangulation injury due to a coil of hair acting as a tourniquet is an uncommon injury in boys that has severe potential complications if diagnosis is delayed. We report the first such case in an uncircumcised boy. The purpose of this report is to call attention to hair as a potential source of injury in the pediatric population. The pathophysiology and management of these injuries are presented. PMID- 12735814 TI - Total endoscopic management of the encrusted ureteral stent under a single anaesthesia. AB - OBJECTIVE: Stented patients lost to follow up may return with large stone burdens encrusting the stent. The study describes total endoscopic management under one anaesthesia to remove such stents. METHODS: A similar approach was done in all cases. Cystoscopic Holmium: YAG (Ho) or electrohydraulic lithotripsy (EHL) removed the bladder calculus. The bladder portion of stent was cut with endoscopic scissors and removed. The patient was repositioned prone for percutaneous access and nephrolithotomy (PCNY), using either ultrasound, EHL, or Ho. The remaining cephalic portion of stent was removed percutaneously. A universal stent was placed and removed one week later. RESULTS: Eleven patients with 12 ureteral stents left indwelling over one year presented with significant stone burden both in the bladder and kidney. Seven patients had failed extracorporeal shock wave lithotripsy (ESWL) prior to referral. One patient with biolateral encrusted stents had both stents treated under one anaesthesia. The average bladder and kidney stone burdens measured 4 and 6 cm, respectively. The median anaesthesia time was 3.5 hours. No patient required transfusion. Ten of eleven patients were rendered stone-free in one procedure. One patient with a complete staghorn calculus was rendered stone-free by post-PCNL ESWL for a residual calyceal stone. CONCLUSIONS: Total endoscopic management of the encrusted ureteral stent accomplishes safe, effective stone and stent treatment under one anaesthetic. Urologists may wish to consider this strategy when ESWL is unlikely to eradicate large stone burdens on encrusted stents. PMID- 12735815 TI - EDITORIAL. PMID- 12735816 TI - The International Society of Urology is holding its 24th triennial congress in Montreal "September 7-11, 1997" PMID- 12735817 TI - Can radiotherapy salvage isolated local recurrence following radical prostatectomy? AB - The optimal management of isolated local recurrence of prostate cancer following radical prostatectomy remains unknown. Salvage radiotherapy to the prostatic bed is one of management options. This article reviews the efficacy of salvage radiotherapy. The available data suggest that salvage radiotherapy offers excellent clinical local control. However, the probability of long-term disease free survival with salvage radiotherapy alone appears small as a significant proportion of patients develop distant metastases and rising prostate specific antigen (PSA) following radiotherapy. A strategy of combining systemic therapy with local salvage treatment needs to be explored for this poor prognostic group. PMID- 12735818 TI - Performance characteristics of a new monoclonal antibody test for bladder cancer: ImmunoCyt trade mark. AB - Transitional carcinoma of the bladder, an important tumor in both men and women, has traditionally been diagnosed and monitored using either or both of two different methodologies. Urinary cytology is highly specific, but has poor sensitivity. Cystoscopy with biopsy is highly accurate, but invasive. A newly developed immuno cytochemical test, ImmunoCyt trade mark (DiagnoCure Inc.), uses three monoclonal antibodies directed against transitional cell carcinoma antigens in exfoliated cells. This study involved 102 asymptomatic controls, and 198 patients with confirmed bladder cancer, in an assessment of the performance characteristics of ImmunoCyt. Combining this test with conventional urinary cytology improves accuracy. Depending on the cutoff level used in the interpretation of a positive result, the highest sensitivity of the test was 95%, with a corresponding specificity of 76.5%. This procedure offers the possibility of non-invasive diagnosis and monitoring of bladder cancer, with acceptable performance characteristics. PMID- 12735819 TI - Parameters predictive of multicentricity in renal cell carcinoma. AB - Over a 3-year period, 83 radical nephrectomies were performed for renal cell carcinoma. The preoperative radiologic investigations and pathology were reviewed. The incidence of multicentricity of the tumors was correlated with various clinical and pathologic parameters. These included polar location, histologic type, histologic grade (divided into low: Fuhrman grade 1 to 2; and high: Fuhrman grade 3 to 4), size on pathology, pathologic stage, capsular invasion, microscopic vascular invasion, renal vein or inferior vena cava (IVC) involvement, collecting system involvement, lymph node metastases and adrenal gland metastases. The incidence of multicentricity was 9/83 (11%). On univariate analysis, no factors were predictive of multicentricity. On logistic regression analysis, microscopic vascular invasion (p=0.021) and tubulo-papillary histology (p=0.049) were the only significant independent predictors of multicentricity. A model incorporating these parameters missed all multicentric tumors. PMID- 12735820 TI - Parameters predictive of multicentricity in renal cell carcinoma - page 406. PMID- 12735821 TI - When is ESWL of small calyceal stones indicated? AB - PURPOSE: To evaluate the indications for and the outcomes of extracorporeal shock wave lithotripsy (ESWL) of calyceal stones <5 mm, we prospectively studied a series of 25 patients scheduled for treatment. MATERIALS AND METHODS: Between January 1994 and June 1995, 15 males and 10 females with a mean age of 44 years (median 43; range 4 to 70 years) underwent ESWL of calyceal stones <5 mm using an unmodified HM3 Dornier lithotriptor. RESULTS: Indications for ESWL included 17 patients (68%) suffering from back and flank pain, 3 patients (12%) with gross hematuria and 1 patient (4%) with recurrent episodes of pyelonephritis. Six patients (24%) were asymptomatic. Evaluation 3 months after ESWL revealed that of the 17 patients with flank pain, 13 (76%) had complete resolution of pain and 4 (24%) had partial resolution of pain and there had been no further episodes of gross hematuria or pyelonephritis in the previously affected patients. At 3-month follow-up, of the 31 kidneys treated, plain radiography were available in 29; 20 (69%) were clear and 9 (31%) had small residual fragments. CONCLUSIONS: Calyceal stones <5 mm in maximum diameter can cause significant symptoms such as pain, hematuria and infection and should not be dismissed as inconsequential. When no other obvious cause is apparent, these small stones should be treated with a high likelihood of resolution of symptoms and clearance. ESWL should not be withheld in patients with small asymptomatic calculi who request or require treatment for fear of unexpected colic or stone growth. PMID- 12735822 TI - Fully hydrated images of Pseudomonas aeruginosa biofilm on the surface of catheter material. AB - We examined the applicability of environmental scanning electron microscopy (ESEM) and confocal laser scanning microscopy (CLSM) for non-invasive imaging of intact bacterial biofilms. Fully hydrated images of Pseudomonas aeruginosa biofilms developing on the silicon disks in an in vitro experimental model were detected simply but clearly by ESEM and CLSM. Three-dimensional spatial images of the bioflims were obtained by ESEM using specimens with neither drying nor conductive coating. Horizontal and vertical sectioning images of the biofilms were obtained by using CLSM in conjunction with positive fluorescence staining techniques; acridine orange and sufranin were successfully used for detection of bacterial cells in the biofilms, while extracellular polymeric substances were obtained selectively with FITC-concanavalin A. The new technology reported here is applicable to diagnostic study as well as fundamental study of biofilm associated infections in the urinary tract. PMID- 12735824 TI - Catheter management related to radical retropubic prostatectomy. AB - The clinical course of 108 consecutive radical retropubic prostatectomy (RRP) patients with regards to postoperative cystograms and suprapubic cystostomy was retrospectively reviewed. All patients had a urethral Foley (UF), suprapubic catheter (SP) and were discharged with both catheters in place. Two of 9 patients who had excessive drainage had biochemical evidence of urine. Cystogram on these 2 patients, deferred till days 19 and 27, was normal. Among the other 7 patients with excessive JP drainage, only 1 showed a minor anastomotic look on day 15, necessitating catheter drainage for 1 additional week, with uneventful Foley removal then. Ten patients showed some degree of extravasation on the initial cystograms, 6 patients were judged to be insignificant and the catheter was removed the same day, 4 patients had extravasation (performed days 7, 13, 15, 22) which warranted repeat cystogram 1 week later, which then turned out to be normal. Five patients experienced problems with the UF early in the postoperative period and the SP served as a backup for bladder drainage. The SP entails negligible morbidity and may serve a useful function in the most critical early postoperative period. Based on our data, routine cystogram prior to catheter removal did not appear to alter the management of the catheters significantly and appears unnecessary, especially if the catheter is left in situ for close to 3 weeks prior to removal. In selected cases where technical anastomotic problems were encountered or if delayed healing was anticipated, a c;ystogram prior to catheter removal would still be available. PMID- 12735823 TI - Proliferating cell nuclear antigen (PCNA) and MIB 1: Markers of locally advanced and biologically aggressive prostate cancer. AB - Using archival radical prostatectomy specimens from patients with prostate cancer (n=19) we studied the relationship between staining with two different markers of cellular proliferation and the local extent or eventual progression of disease. In cases where the disease had spread outside the capsule of the prostate (n=9) the average staining patterns for proliferating cell nuclear antigen (PCNA) and MIB 1 were higher than in cases where the disease was organ-confined (n=10). The difference in the staining patterns for these two groups was greater for the MIB 1 antigen and this was shown to be statistically significant (p=0.01) whereas the smaller difference seen for proliferating cell nuclear antigen was not statistically significant. There was a higher staining pattern seen for these antigens in cases where disease progression was evident by a rising prostatic specific antigen level. The difference was greater and statistically significant (p23%). No differences in marginal adaptation were detected for both application techniques if the same adhesive was used (ANOVA, Tukey, p<0.05). Lining with Revolution resulted in a significantly higher percentage of marginal continuity (MC) and less enamel fractures (EF) compared to Tetric Flow (MC: 73 vs. 45%; EF: 23 vs. 50%). Gaps at the inner interface were rarely observed (<0.05%). CONCLUSIONS: Large Class I restorations without dentin supported cavity margins showed a high amount of marginal enamel fractures directly after placement. Lining with Revolution proved to have the highest potential to initially maintain the marginal integrity. PMID- 12735917 TI - The erosive potential of some herbal teas. AB - OBJECTIVES: Many epidemiological studies show a high prevalence of tooth wear, even in young patients. One factor that may be contributing to this problem is the consumption of herbal teas that are often considered to be 'healthy' alternatives to other beverages. The aim of this study was to screen a number of these products for their potential to cause erosion. METHODS: The erosive potential of a variety of herbal teas was assessed in the laboratory by measuring their pH, neutralisable acidity and their ability to erode enamel and these were compared to a positive control, orange juice. RESULTS: The pH of the herbal teas ranged from 3.1 to 7.1 and the neutralisable acidity ranged from 3.5 to 60.3 ml of 0.1M NaOH. The amount of enamel removed following 1h immersion in the herbal teas ranged from 0.00 to 9.6 microm. In comparison, the orange juice control had a pH of 3.7 a neutralisable acidity of 21.4 ml and removed 3.3 microm of enamel. CONCLUSIONS: Many of the herbal teas tested were found to be more erosive than orange juice. This information will be of use to clinicians when counseling patients with tooth surface loss. PMID- 12735918 TI - Effect of circumferential undercut placement in the root canal wall on the retention of a tapered post in large and small root canals. AB - OBJECTIVES: This study examined the influence of root canal space and circumferential undercut placement in the dentine wall on the retention of a prefabricated Root Keeper post. METHODS: Twenty-four extracted single-rooted teeth, having narrow root canal spaces, were decoronated, treated endodontically, and then centrally embedded in resin blocks. All canal spaces were prepared with a no. 2 Peeso reamer to an 8mm depth, and half the specimens were successively treated with a no. 3 Peeso to depths of 5 and 8mm for the small canal group (SC) and large canal group (LC), respectively. An L-type keeper post (Aichi Steel Co.) was luted to each canal with resin cement. The assembly was thermocycled 2000 times (4-60 degrees C), and post retention was measured. Afterwards the dislodged post with the intact structure was sandblasted, and the canal was cleaned and slightly undercut using an inverted cone bur. Six treated posts were recemented for both grooved large canal (GLC) and grooved small canal (GSC) groups, and the repaired retentive strengths were measured. RESULTS: Keeper posts cemented in SC (84.26+/-31.85N) showed better (p<0.05) retention than those cemented in LC (41.92+/-22.36N). Interestingly, the repaired strengths of the recemented post increased in both GSC (104.29+/-11.89N) and GLC (96.65+/-6.91N). All posts, except for one in the GLC, failed at the keeper-post joint. CONCLUSIONS: Prefabricated Root Keeper posts had lower retentive strength in roots with a large canal space. Placement of a circumferential undercut in the root canal wall significantly enhanced the retention of the short tapered posts. PMID- 12735920 TI - Micro-mechanical characterisation of the properties of primary tooth dentine. AB - OBJECTIVES: Understanding the mechanical properties of dentine is of importance as adhesive restorative materials mainly achieve their bonding to the tooth structure through dentine. The current study measures the hardness and modulus of elasticity of primary molar dentine using an Ultra-Micro-Indentation System (UMIS), which allows the dentine to remain hydrated and thus is assumed to be closer to the in vivo conditions. METHODS: Eight sound primary molar teeth were axially sectioned, embedded in resin and fine polished. Two linear arrays of indentations were done on coronal dentine, from the pulp wall to dentino-enamel junction (DEJ) parallel to the tubule direction under a force load of 25mN. RESULTS: The mean hardness and elastic modulus of the dentine nearest the pulp wall was 0.52+/-0.24 and 11.59+/-3.95GPa, respectively, which was significantly lower than those of dentine in the middle area, which was 0.85+/-0.19 and 17.06+/ 3.09GPa, respectively, and the dentine nearest DEJ, which was 0.91+/-0.15 and 16.33+/-3.83GPa, respectively. There is a statistically significant linear correlation between the hardness and modulus of elasticity. CONCLUSIONS: The hardness and modulus of elasticity of dentine decreases with decreasing distance from the pulp. This is of importance to clinicians because an extension of cavity preparation towards the pulp may lead to less mechanical support for a restoration. PMID- 12735919 TI - Development of low erosive carbonated fruit drinks. 1. Evaluation of two experimental orange drinks in vitro and in situ. AB - OBJECTIVES: To determine the in vitro erosive potential and in situ erosive effect of two new formulation low calorie carbonated orange drinks with that of two conventional diet products and water. METHODS: In the in vitro study, six specimens of deciduous and permanent enamel were randomly allocated to each of the five products and a '4h' protocol employed. In the in situ study, 15 healthy volunteers participated in a single centre, single blind, 5-phase crossover study, conducted according to Good Clinical Practice, and employing a validated model. RESULTS: The in vitro erosive potential of the experimental formulations was less than that of the comparators at all time points. Conversely, the observed erosive potential of both experimental formulations was greater than that of the control. Consistent statistically significant differences were found in relation to permanent enamel only. Unfortunately, the in situ study did not produce results entirely consistent with those of the in vitro study. Notably, a generally progressive loss of enamel was observed in specimens exposed to the control. CONCLUSIONS: The data from the in vitro study show the experimental formulations to have low comparative erosivity. However, the methodologies in vitro and in situ somewhat unusually do not correlate in ranking the erosivity of drinks. The results of this study should therefore be viewed with caution, further research being clearly warranted. PMID- 12735921 TI - A new polyisoprene-based light-curing denture soft lining material. AB - OBJECTIVES: The purpose of this study was to investigate some clinically relevant properties of Clearfit LC, a newly developed polyisoprene-based light-curing lining material. Its properties were compared with those of other four commercial products. MATERIALS AND METHODS: Five soft lining materials, Clearfit LC, two plasticised acrylics (Super-soft, Soften) and two silicones (Molloplast B, Sofreliner) were used to determine water sorption, solubility, staining resistance, Shore A hardness and shear bond strength to denture base. Five specimens for each test were fabricated. The results of each test among the five materials were compared by one-way ANOVA and Scheffe's post-hoc test at alpha=0.05. RESULTS: One-way ANOVA of each test showed that the differences among five materials were significant (P<0.05). Clearfit LC showed the intermediate water sorption and solubility (10.07; 1.72 microg/mm(3)). It also showed the lowest color change in beta-carotene/olive oil solution and instant coffee solution (DeltaE=1.19, 5.48, respectively). The shear bond strength of Clearfit LC was 1.56MPa, and no adhesive failure was found. Its Shore A hardness value (56.2) was located between two acrylics and silicones. It was also found that the softer materials tend to failure cohesively. CONCLUSIONS: The polyisoprene-based lining material showed low water sorption and solubility, moderate softness, high staining resistance and satisfactory shear bond strength to denture base resin. It also provides clinicians sufficient working time due to its light-curing property. It would be an attractive alternative as a relining material. PMID- 12735922 TI - Bacterial penetration through canals of endodontically treated teeth in the presence or absence of the smear layer. AB - OBJECTIVES: The goal of this study is to determine if the smear layer affects the passage of bacteria through or around obturating material as evidenced by penetration of bacteria through and out the canal. Specifically, this study focused on determining the effect of the smear layer on the magnitude of bacterial penetration through the apical foramen. METHODS: Thirty extracted, maxillary central or lateral incisors were collected. Teeth were randomly assigned (10 teeth per group) to three groups: (1) smear layer removed, (2) smear layer present, and (3) negative control. Canal preparation and obturation using lateral condensation, gutta percha, and AH 26 sealer was performed on all of the teeth. Removal of the smear layer was accomplished by rinsing with 17% EDTA. The model systems consisted of an upper chamber attached to the cemento-enamel junction and a lower chamber at the apices of the teeth. Standardized bacterial suspensions containing Fusobacterium nucleatum, Campylobacter rectus, and Peptostreptococcus micros were inoculated into the upper chambers. Models were incubated anaerobically at 37 degrees C. At various times over a 60-day period, samples were taken from the lower chamber and spiral-plated on selective differential media to determine numbers and types of bacteria. RESULTS: Leakage results were as follows: (1) smear layer present-6/10 leaked; (2) smear layer removed-0/10 leaked; (3) negative control-0/10 leaked. Profiles of bacterial leakage were similar among the groups. F. nucleatum was the predominant microorganism. CONCLUSIONS: This study indicated that removal of the smear layer reduced the leakage of bacteria through the root canal system. PMID- 12735923 TI - Evaluation of mutans streptococci in plaque and saliva: correlation with caries development in preschool children. AB - OBJECTIVES: To test the ability of a site-specific method of evaluating mutans streptococci (MS) present in plaque in order to predict caries risk in children and to compare the results with those obtained from estimating the levels of MS present in saliva. METHODS: Caries prevalence was examined twice, 6 months apart, in 129 preschool children ranging in age from 1.5 to 5. A commercial strip method was used to evaluate the MS score (0-3) in plaque collected from four selected tooth surfaces at two interdental spaces and in saliva. RESULTS: The screening criteria of the MS score for caries incidence between scores 1 and 2, and scores 0 and 1 were the most valid for evaluating plaque (accuracy of prediction, 80%) and saliva (accuracy of prediction, 71%), respectively. Based on these screening criteria, the evaluation of saliva MS was invalid when the plaque MS score was considered. Screening plaque MS was effective, regardless of caries experience at baseline, according to the chi-square test (p<0.01), but screening saliva MS was ineffective when caries experience was present at baseline. A multivariate logistic regression analysis including six variables showed that the plaque MS score and caries experience at baseline were significantly correlated with caries incidence: caries experience at baseline (OR=5.02, 95%CI: 1.81-14.59) and high plaque MS score (2 or 3) (OR=12.59, 95%CI: 3.18-67.08). CONCLUSIONS: These results suggest that the semi-quantitative evaluation of plaque MS sampled from only two proximal sites on the molars is more effective for screening preschool children at a high risk of developing caries in their primary dentition than other important caries risk indicators. PMID- 12735924 TI - The effect of magnification on the iatrogenic damage to adjacent tooth surfaces during class II preparation. AB - OBJECTIVES: To determine the amount of damage to adjacent surfaces during cavity preparation of approximal box-cavities by using either no magnification or an individually adapted surgical telescope. MATERIALS AND METHODS: Nine dentists prepared in a mannequin head 4 approximal (class II) box-cavities without magnification using a high speed handpiece with lighting from a dental unit. At least 2 months later the test was repeated with a surgical telescope system with integrated light. The degree of damage the 72 adjacent surfaces suffered was determined. RESULTS: The average time needed to prepare one cavity was 14.9min without and 18.3min with magnification (p=0.01). Altogether, 29.7% of adjacent area were damaged when no surgical telescopes were used and 34.5% when surgical telescopes were applied (p>0.05). When the mesial-facing and distal-facing surfaces were analysed independently a statistically significant increase of damage was found on distal surfaces (p=0.03) when using surgical telescopes. Seventy surfaces (=97%) had a preparation trauma. CONCLUSIONS: Surgical telescopes do not decrease damage of adjacent tooth surfaces. PMID- 12735925 TI - Effect of glass fibre reinforcement on residual methyl methacrylate content of denture base polymers. AB - OBJECTIVES: Residual methyl methacrylate (MMA) content in unreinforced and in glass fibre reinforced, heat-polymerized (long- and short-term terminal boiled and conventionally) autopolymerized (at room temperature and in water at 60 degrees C) and microwave-polymerized (3min at 500W) denture base polymers after processing were compared. METHODS: Ten specimens were prepared for each curing cycle (five unreinforced and five reinforced) adding up to a total of 60. Residual MMA content was determined using high-performance liquid chromatography. Data were analysed with one-way ANOVA followed by Tukey HSD and Paired Samples tests. RESULTS: For unreinforced and reinforced groups; residual MMA content succesively ranked from lowest to highest in; long- and short-term terminal boiled heat-polymerized, microwave-polymerized, autopolymerized specimens processed in water at 60 degrees C and conventionally heat- and autopolymerized specimens processed at room temperature. Generally residual MMA was found more in glass fibre reinforced test groups than unreinforced groups. However, when reinforced residual MMA increased significantly in long- and short-term terminal boiled heat- (P<0.05) and microwave-polymerized test groups (P<0.01). CONCLUSIONS: Although this increase was significant, lowest residual MMA content was found succesively in reinforced long- and short-term terminal boiled heat polymerized and microwave-polymerized like in unreinforced groups. PMID- 12735927 TI - Effects of growth hormone on fluid homeostasis. Clinical and experimental aspects. PMID- 12735928 TI - Measuring IGF-I, IGFBP-2 and IGFBP-3 from dried blood spots on filter paper is not only practical but also reliable. AB - OBJECTIVE: The aim of this study is to adapt measurements of IGF-I, IGFBP-2 and IGFBP-3 to dried blood filter disk assays. METHODS: Measurements of the three analytes in serum samples and in the corresponding blood spotted onto filter paper were compared by applying standard radioimmunoassays. RESULTS: In paired experiments, the quantity of all antigens measured on dried filter spots showed an excellent correlation to that in serum (R>0.88) and in addition this correlation was independent of the whole blood hematocrit value. Recovery of IGF I, IGFBP-2 and IGFBP-3 in experiments using recombinant standards mixed with whole blood was well correlated with the recovery of the plasma fraction on the filter paper. All of the blood spot assays showed a inter- and intra-assay variation of less than 10% and the blood spots were stable over a period of more than 5 months stored at -20 degrees C. CONCLUSIONS: Taken together, these data clearly demonstrate that such a filter paper assay is a reliable procedure to monitor changes of IGF-I, IGFBP-2 and IGFBP-3 content in blood. The obvious advantages of this methods concerning storage, handling and shipping of blood probes helps to solve the logistics of centralised measurements of these three analytes. PMID- 12735929 TI - Effects of growth hormone replacement therapy on metabolic and cardiac parameters, in adult patients with childhood-onset growth hormone deficiency. AB - OBJECTIVE: We evaluated metabolic and cardiac parameter changes with GH-therapy. DESIGN: Sixteen adults with childhood-onset hypopituitarism receiving pituitary hormone replacement, except GH-replacement, were assessed at baseline and after 6 and 12 months of GH-replacement. Sixteen healthy adults matched for sex, age, weight, height, body mass index, and body surface area served as the control group to compare cardiac function in both groups. RESULTS: All patients had GH deficiency. After 12 months, serum insulin-like growth factor-1 levels normalized. Basal glucose or insulin levels had no alterations. The low/high density lipoprotein-cholesterol ratio decreased (3.18+/-1.32 x 2.17+/-0.8, p<0.001). Percent lean body mass increased (69.9+/-5.5 x 78.4+/-8.1%), and percent fat body mass decreased (30.1+/-5.5 x 21.6+/-8.1%) (both, p<0.001). Before treatment, patients had decreased left ventricular (LV) echocardiographic morphologic indexes, which were corrected (initial versus 12 months): interventricular septal thickness (0.68+/-0.06 x 0.78+/-0.06 cm), LV posterior wall thickness (0.69+/-0.07 x 0.78+/-0.05 cm), and LV mass index (58.9+/-11.0 x 71.1+/-9.4 g/m(2)) (all, p<0.001). Exercise capacity improved, as assessed by oxygen consumption (7.84+/-1.44 x 9.67+/-1.74 METS, p<0.001). CONCLUSIONS: GH replacement seems to reduce cardiovascular risks in adults with childhood-onset GH-deficiency. PMID- 12735930 TI - Type Ialpha collagen is an IGFBP-3 binding protein. AB - Insulin-like growth factor (IGF) binding protein-3 (IGFBP-3) possesses both growth-inhibitory and -potentiating effects on cells, which are independent of IGF action and mediated through specific IGFBP-3 binding proteins/receptors located at the cell membrane, cytosol, or nuclear compartments as well as in the extracellular matrix. We here characterized type Ialpha collagen as one of these IGFBP-3 binding proteins. Human serum was fractionated over an IGFBP-3 affinity column, and bands at 70-100 kDa were eluted as IGFBP-3 ligands. The 100-kDa band was extracted, subjected to N-terminal amino acid sequencing, and identified through database searching as the N-terminal chain of type Ialpha collagen protein. In a separate screening approach, using a yeast two-hybrid system, we cloned the type Ialpha collagen cDNA from a human liver cDNA library as an IGFBP 3 protein partner. Anti-IGFBP-3 antibodies co-immunoprecipitated type Ialpha collagen and IGFBP-3 from the conditioned media of human fibroblasts and vice versa. We demonstrated through ligand dot blot analysis that type Ialpha collagen binds IGFBP-3. IGFBP-3 mutants, with altered sequence at the nuclear localization sequence, bound type Ialpha collagen poorly. Western immunoblot showed that type Ialpha collagen binds only IGFBP-3 but not IGF-I, suggesting an IGF-I-independent mechanism of this interaction. Physiological effects of IGFBP-3-collagen interactions may include modulation of cell adhesion and migration. PMID- 12735931 TI - Biochemical definitions of disease activity in acromegaly. AB - In acromegaly the therapeutic outcome is difficult to assess and depends on the biochemical method. We have ascertained disease activity in 70 acromegalic patients by means of a GH profile (8 hourly samples) and a single IGF-I measurement as compared to a healthy control group. As an estimate of the "stiffness" of the GH profile we calculated the SD/nadir(GH) from the GH profile. In the control group the following upper normal limits were obtained: IGF-I (microg/l) 217; mean GH (microg/l) 2.16; nadir GH (g/l) 0.3. Based on ROC plot analysis a value of 2.0 for the SD/nadir ratio was used as cut-off. This translated into the following surgical cure rates (%): IGF-I 47; mean GH 77; nadir GH 65; SD/nadir 30. Some of the patients post-surgery had elevated IGF-I levels despite "normal" GH levels. Abnormal SD/nadir versus normal IGF-I and vice versa were recorded in many patients post-surgery. IN CONCLUSION: (1) cure rates of acromegaly depend strongly on the criteria being used and (2) estimates of GH secretion pattern may yield important information about GH status in acromegaly. PMID- 12735932 TI - Occurrence of GH deficiency in adult patients who underwent neurosurgery in the hypothalamus-pituitary area for non-functioning tumour masses. AB - Hypothalamus-pituitary tumours and their treatments (neurosurgery and/or radiotherapy) are major causes of acquired hypopituitarism. Scientific and clinical evidences show the positive effect of GH replacement therapy in severe adult GH deficiency (GHD) pointed toward the need of diagnostic screening of conditions at high risk for GHD. We screened 152 adults (82 males, 70 females; age: 52.3+/-1.2 years, age-range: 20-80 years, BMI: 26.4+/-0.8 kg/m(2)) in order to disclose the presence of GHD after neurosurgery for hypothalamus-pituitary tumours. The whole group (studied at least 3 months after neurosurgery) included: 111 non-functioning pituitary adenomas and 41 peri-pituitary tumours (24 craniopharyngiomas, 7 meningiomas, 5 cysts, 2 chondrosarcomas, 1 colesteatoma, 1 germinoma and 1 hemangiopericitoma). In 14 patients who underwent both neurosurgery and radiotherapy due to a tumour remnant, the somatotroph function was evaluated again 6 months after the end of radiotherapy. GHD was assumed to be shown by GH peak <5 microg/L (severe <3 microg/L) after Insulin Tolerance Test (ITT) or <16.5 microg/L (severe <9 microg/L) after GH-releasing hormone+arginine test (GHRH+ARG) (3rd and 1st centile limits of normality, respectively), two widely accepted provocative tests. Before neurosurgery GHD was present in 97/152 (63.8%) and resulted severe in 66/152 (43.4%) patients. After neurosurgery GHD was present in 122/152 (80.2%) and severe in 106/152 (69.7%). While 26 patients developed severe GHD (GHD) as consequence of neurosurgery, only one patient who had been classified as GHD before neurosurgery showed normal GH response after surgery. After neurosurgery, 91.0% (81/89) of the pan-hypopituitaric patients showed severe GHD. Considering the 14 patients who underwent also radiotherapy after neurosurgery, 7/14 had GHD before neurosurgery while 12/14 became severe GHD after radiotherapy in a context of pan-hypopituitarism. IGF-I levels below the 3rd age-related normal limits were present in 39.0% of patients in whom severe GHD was showed by provocative tests. In conclusion, this study shows that the occurrence of acquired severe GHD is extremely common in adult patients bearing non-functioning tumour masses in the hypothalamus-pituitary area and further increases after neurosurgery. All patients bearing non-functioning hypothalamus-pituitary tumours should undergo evaluation of their somatotroph function before and after neurosurgery that represents a condition at obvious more than high risk for hypopituitarism. PMID- 12735933 TI - Cortical auditory disorder caused by bilateral strategic cerebral bleedings. Analysis of two cases. AB - The authors present the anatomical and clinical features of cortical auditory dysfunction in two patients, in whom hypertensive bleedings destroyed the cortical auditory centres in both hemispheres. The second haemorrhage occurred four years after the first bleeding in both cases. The bleedings isolated the cortical hearing centres due to the destruction of the right and left temporal white matter. During the clinical course the symptomatology has changed in both patients: pure word deafness has transformed to cortical deafness and generalised auditory agnosia. Normal pattern of brain stem auditory evoked potentials suggested intact auditory pathways. Authors conclude that transformation of clinical forms of cortical auditory disorder can be explained by the tissue reaction to the subcortical bleeding in the cortical auditory centres. PMID- 12735934 TI - Speed of lower-level auditory and visual processing as a basic factor in dyslexia: electrophysiological evidence. AB - This study investigated speed of processing (SOP) among college-level adult dyslexic and normal readers in nonlinguistic and sublexical linguistic auditory and visual oddball tasks, and a nonlinguistic cross-modal choice reaction task. Behavioral and electrophysiological (ERP) measures were obtained. The results revealed that between both groups, reaction times (RT) were longer and the latencies of P2 and P3 components occurred later in the visual as compared to auditory oddball tasks. RT and ERP latencies were longest in the cross-modal task. RT and ERP latencies were delayed among dyslexic as compared to normal readers across tasks. On the oddball tasks, group differences in brain activity were observed only when responding to low-probability targets. These differences were largest for the P3 component, and most pronounced in the case of phonemes. The gap between ERP latencies in the visual versus the auditory modalities for each component was larger among dyslexic as compared to normal readers, and was particularly evident at the linguistic level. A hypothesis is proposed that suggests an amodal, basic SOP deficit among dyslexic readers. The slower cross modal SOP is attributed to slower information processing in general and to disproportionate "asynchrony" between SOP in the visual versus the auditory system. It is suggested that excessive asynchrony in the SOP of the two systems may be one of the underlying causes of dyslexics' impaired reading skills. PMID- 12735936 TI - Effect of bilateral stimulation of the subthalamic nucleus on parkinsonian dysarthria. AB - Never was the effect of bilateral stimulation of the subthalamic nucleus (STN) evaluated quantitatively on all the components of speech production, that is articulation, respiration and phonation, at the same time. It is the purpose of this study which uses force measurements of the articulatory organs and acoustic analysis in 16 parkinsonian patients. With STN stimulation, reaction and movement time of the articulatory organs decreased and their maximal strength, as well as their precision increased. We also noted a large beneficial effect on voice with a significant improvement in respiratory and phonatory functions. PMID- 12735935 TI - Thalamic stuttering: a distinct clinical entity? AB - A 38-year-old right-handed male with no history of speech or language problems presented with neurogenic stuttering following an ischaemic lesion of the left thalamus. He stuttered severely in propositional speech (conversation, monologue, confrontation naming, and word retrieval) but only slightly in non-propositional speech (automatic speech, sound, word and sentence repetition, and reading aloud). It is suggested that thalamic stuttering may constitute a distinct clinical entity. PMID- 12735937 TI - Facilitatory and inhibitory effects of grammatical agreement: evidence from readers' eye fixation patterns. AB - The study examined how grammatical agreement affects reading in Finnish. Readers' eye fixation patterns were recorded when they read one of three alternative versions of the same sentences, where the critical difference was the type of preceding word of the target nouns. The preceding word was (a) an agreeing modifier (mainioksi orkesteriksi='for an excellent orchestra'), (b) a non agreeing modifier that was grammatical, unambiguous and synonymous to the agreeing modifier (kelpo orkesteriksi='for an excellent orchestra'), or (c) a baseline condition without a modifier (orkesteriksi='for an orchestra'). Two different types of agreement were used, a modifier-head agreement and a possessive agreement. The results showed that the agreeing modifiers facilitate and the non-agreeing modifiers inhibit the reading of the target nouns compared to the neutral baseline condition. These effects appeared in the second-pass reading. The pattern was similar between the two agreement structures. PMID- 12735938 TI - Characterizing the time course of an implicature: an evoked potentials study. AB - This work employs Evoked Potential techniques as 19 participants are confronted with sentences that have the potential to produce scalar implicatures, like in Some elephants have trunks. Such an Underinformative utterance is of interest to pragmatists because it can be considered to have two different truth values. It can be considered true when taken at face value but false if one were to treat Some with the implicature Not All. Two accounts of implicature production are compared. The neo-Gricean approach (e.g., Levinson, 2000) assumes that implicatures intrude automatically on the semantics of a term like Some. Relevance Theory (Sperber & Wilson, 1985/1996) assumes that implicatures are effortful and not automatic. In this experiment, the participants are presented with 25 Underinformative sentences along with 25 sentences that are Patently True (e.g. Some houses have bricks) and 25 that are Patently False (e.g. Some crows have radios). As reported in an earlier study (Noveck, 2001), Underinformative sentences prompt strong individual differences. Seven participants here responded true to all (or nearly all) of the Underinformative sentences and the remaining 12 responded false to all (or nearly all) of them. The present study showed that those who responded false to the Underinformative sentences took significantly longer to do so that those who responded true. The ERP data indicate that: (a) the Patently True and Patently False sentences prompt steeper N400's--indicating greater semantic integration--than the Underinformative sentences and that (b) regardless of one's ultimate response to the Underinformative sentences, the N400's were remarkably flat, indicating no particular reaction to these sentences. Collectively, the data are taken to show that implicatures are part of a late-arriving, effort-demanding decision process. PMID- 12735939 TI - Neural basis for sentence comprehension deficits in frontotemporal dementia. AB - Many patients with frontotemporal dementia (FTD) have impaired sentence comprehension. However, the pattern of comprehension difficulty appears to vary depending on the clinical subgroup. The purpose of this study was to elucidate the neural basis for these deficits in FTD. We studied patients with two different presentations: Three patients with Progressive Non-Fluent Ahasia (PNFA), and five non-aphasic patients with a dysexecutive and social impairment (EXEC). The FTD patient subgroups were compared to a cohort of 11 healthy seniors with intact sentence comprehension. We monitored regional cerebral activity with blood oxygen level dependent (BOLD) functional magnetic resonance imaging (fMRI) while subjects read sentences featuring both a grammatically complex object relative center-embedded clause and a long linkage between the head noun phrase (NP) and the gap where the NP is interpreted in the center-embedded clause. Subjects decided whether the agent of the action is a male or a female. Healthy seniors activated both ventral portions of inferior frontal cortex (vIFC) and dorsal portions of IFC (dIFC) in the left hemisphere, often associated with grammatical and working memory components of these sentences, respectively. PNFA patients differed from healthy controls since they have reduced activation of left vIFC, while EXEC patients have less recruitment of left dIFC. We conclude that FTD subgroups have distinct patterns of sentence comprehension difficulty in part because of selective interruptions of a large-scale neural network for sentence processing. PMID- 12735940 TI - Demonstrating a wordlikeness effect on nonword repetition performance in a conduction aphasic patient. AB - The purpose of this study was to identify the nature of the deficit for a conduction aphasic patient in order to evaluate two different theories of conduction aphasia. First, a conduction aphasic patient FS was tested on auditory word-pair discrimination, word-repetition, and picture-naming. The results of these tasks indicated that her deficit was likely to be post-lexical rather than perceptual or lexical. Next, we examined her repetition performance for two types of nonwords (high-wordlike and low-wordlike nonwords) to distinguish the two theories. FS exhibited a wordlikeness effect: she produced more correct moras and more correct combinations of moras for high-wordlike nonwords than low-wordlike nonwords. We conclude that she had difficulty in maintaining stable phonological representations of verbal materials in the output buffer. PMID- 12735941 TI - Event-related brain potential markers of high-language proficiency in adults. AB - The central issue addressed is whether there are electrophysiological markers for high-language proficiency in adults. Event-related brain potentials (ERPs) were recorded in 19 young adults who demonstrated either "normal" or "high" language proficiency. ERPs were obtained during a sentence-processing task and analyzed for specific response components elicited by different word types. The ERPs in the normal and high-language proficiency groups were similar for early sensory related processing but differed in the later components thought to be more closely related to lexical access and post-lexical processing. The findings for the ERP waveforms suggest that adults with high-language proficiency show slightly faster lexical access over left anterior brain regions for closed class items, and a reduced reliance on contextual information contained in sentences. PMID- 12735943 TI - Considerations on agraphia in light of a new observation of pure motor agraphia. PMID- 12735942 TI - PET activation studies comparing two speech tasks widely used in surgical mapping. AB - "Automatic" speech, especially counting, is frequently preserved in aphasia, even when word production is severely impaired. Although brain sites and processes for automatic speech are not well understood, counting is frequently used to elicit fluent speech during preoperative and intraoperative cortical mapping for language. Obtaining both behavioral and functional brain imaging measures, this study compared counting with a word production task (generation of animal names), including non-verbal vocalizations and quiet rest as control states, in normal and aphasic subjects. Behavioral data indicated that normal and aphasic groups did not differ in counting or non-verbal vocalizations, but did differ significantly in word production ("naming" animals). Functional brain imaging results on normal subjects using partial least squares analysis of PET rCBF images revealed three significant latent variables (LVs): one for naming and vocalizing, identifying bilateral anterior areas, with left predominating over right; a second LV for naming, identifying left and right frontal and temporal areas. For the third, only marginally significant LV, which was associated with automatic speech alone (counting), right and subcortical sites predominated. For patients, two LVs emerged, identified with naming and vocalization, and corresponding to a variety of cerebral sites; the analysis failed to find a specific latent variable for counting. A comparison between group data for normal subjects and patients suggested that the naming, counting, and vocalization tasks were performed differently by the two groups. These results suggest that word generation as a verbal task is more likely to elicit activity in classical language areas than counting. Further studies are suggested to better understand differences between neurological substrates for non-propositional and automatic speech. PMID- 12735944 TI - The history of written language disorders: Reexamining Pitres' case (1884) of pure agraphia. AB - The first clinical description of pure agraphia was reported by the French neurologist Pitres in 1884. Pitres used the case study evidence to argue for modality-specific memory representations and the localization of writing. This article reviews Pitres's contribution to the study of acquired writing disorders, the components of writing models and the cerebral localization which subserve writing, in light of the views entertained by his contemporaries and current authors. Although numerous cases have been reported throughout this century, the view that writing can be impaired while other language functions and motor activities remain intact is still challenged. PMID- 12735945 TI - Differences in the perception and time course of syntactic and semantic violations. AB - A reading time and an ERP experiment conducted in Italian investigated the parser's responses to a syntactic violation (subject-verb number agreement) and to a semantic violation (subject-verb selectional restriction), examining the time course of comprehension processes until sentence end. The reading-time data showed that the syntactic violation was detected earlier than the semantic one and that the two violations differed in the time-course. The ERP data fully supported the reading time data: Syntactic anomalies elicited a left anterior negativity (LAN) and a P600. Semantic anomalies elicited a N400 centred on the parietal sites which started 90 ms later (latency 430 ms) than the LAN. Furthermore, the N400 evoked by the words that followed the target word continued and increased until sentence end. The results are discussed with respect to the hypotheses that the parser constructs distinct syntactic and semantic analyses of a sentence and that this characteristic holds cross-linguistically. The appropriateness of different methodologies to the study of sentence processing is also evaluated. PMID- 12735946 TI - Cortical operational synchrony during audio-visual speech integration. AB - Information from different sensory modalities is processed in different cortical regions. However, our daily perception is based on the overall impression resulting from the integration of information from multiple sensory modalities. At present it is not known how the human brain integrates information from different modalities into a unified percept. Using a robust phenomenon known as the McGurk effect it was shown in the present study that audio-visual synthesis takes place within a distributed and dynamic cortical networks with emergent properties. Various cortical sites within these networks interact with each other by means of so-called operational synchrony (Kaplan, Fingelkurts, Fingelkurts, & Darkhovsky, 1997). The temporal synchronization of cortical operations processing unimodal stimuli at different cortical sites reveals the importance of the temporal features of auditory and visual stimuli for audio-visual speech integration. PMID- 12735947 TI - Communicative deficits in agenesis of the corpus callosum: nonliteral language and affective prosody. AB - While some individuals with agenesis of the corpus callosum can perform normally on standardized intelligence tests, clinical observations suggest that they nevertheless have deficits in the domains of fluid and social intelligence. Particularly important for social competence is adequate understanding and use of paralinguistic information. This study examined the impact of callosal absence on the processing of pragmatic and paralinguistic information. Young adult males with agenesis of the corpus callosum (ACC) were evaluated in the areas of nonliteral language comprehension, proverb recognition and interpretation, and perception of affective prosody. Ten ACC individuals with normal Wechsler IQ were compared to 14 sex, age, and IQ matched normal controls. The Formulaic and Novel Language Comprehension Test (FANL-C), Gorham Proverbs Test, and LA Prosody Test were administered. ACC subjects exhibited significant impairment on the nonliteral items of the FANL-C, but no significant difference from controls in comprehension of literal items. ACC subjects also exhibited significant deficits in both self-generated interpretation and recognition of proverb meaning, and in recognition of affective prosody. These results demonstrate that normally intelligent individuals with ACC are impaired in the understanding of nonliteral language and emotional-prosodic cues that are important in social communication. In all three tests, the performance of individuals with ACC was similar to patients with right hemisphere brain damage. Thus, persons with ACC appear to lack interhemispheric integration of critical aspects of language processed by the right hemisphere. PMID- 12735948 TI - The contribution of EEG coherence to the investigation of language. AB - The contribution of EEG coherence analysis to the investigation of cognition and, in particular, language processing is demonstrated with examples of recent EEG studies. The concept of EEG coherence analysis is explained, and its importance emphasized in the light of recent neurobiological findings on frequency-dependent synchrony as a code of information processing between nerve cell assemblies. Furthermore, EEG coherence studies on naturally spoken and written word and sentence processing are reviewed and experimental results are presented giving new insights into the occurrence of "transient functional language centers" within the brain. PMID- 12735949 TI - Coral reef "restoration" and coral transplantation. PMID- 12735950 TI - Data use and information creation: challenges for marine scientists and for managers. AB - In the coastal waters of European countries and in the offshore waters of the north-east Atlantic, there is an increasing need for scientists to meet challenging objectives, such as to identify meaningful measures of 'quality', and to recommend 'indicators' to underpin implementation of directives, conventions, statutes and other more informal national and international initiatives. Those indicators may relate to particular species or habitats, to changes in physical and chemical characteristics, and even to the use to which the system is put. The problems to be overcome are difficult, but new and developing approaches will make a significant contribution. The approaches include: criteria to identify 'sensitivity' and 'importance', structures to organise information and electronic information resources to access data. The real challenge is to make the results of the various scientific initiatives relevant to and understandable by a wide range of customers with similar overlapping requirements, and thus make a genuine contribution to protecting the marine environment. Above and beyond that is the need for scientists to drive the agenda to enable real and lasting progress to be made towards ecosystem-based management of our seas and a proper consideration of what 'sustainability' may mean in the marine environment and how we utilise its resources. PMID- 12735952 TI - Taxonomic sufficiency: an overview of its use in the monitoring of sublittoral benthic communities after oil spills. PMID- 12735951 TI - Biological invasions as a component of global change in stressed marine ecosystems. AB - Biological invasions in marine environment are the lesser known aspect of global change. However, recent events which occurred in the Mediterranean Sea demonstrate that they represent a serious ecological and economical menace leading to biodiversity loss, ecosystem unbalancing, fishery and tourism impairment. In this paper we review marine bioinvasions using examples taken from the Mediterranean/Black Sea region. Particular attention is given to the environmental status of the receiving area as a fundamental pre-requisite for the colonisation success of alien species. The spread of the tropical algae belonging to the genus Caulerpa in the northwestern basin of the Mediterranean Sea has been facilitated by pre-existing conditions of instability of the Posidonia oceanica endemic ecosystem in relation to stress of both natural and anthropogenic origin. Human interventions caused long-term modification in the Black Sea environment, preparing a fertile ground for mass bioinvasion of aquatic nuisance species which, in some cases, altered the original equilibrium of the entire basin. Finally, the Venice lagoon is presented as the third example of an environment subjected to high propagule pressure and anthropogenic forcing and bearing the higher "diversity" of non-indigenous species compared to the other Mediterranean lagoons. Stressed environments are easily colonised by alien species; understanding the links between human and natural disturbance and massive development of non-indigenous species will help prevent marine bioinvasions, that are already favoured by global oceanic trade. PMID- 12735953 TI - Taxonomic sufficiency and the increasing insufficiency of taxonomic expertise. AB - Taxonomic sufficiency (TS) involves the identification of taxa only to a level of taxonomic resolution sufficient to permit the detection of changes in stressed assemblages. Recently, however, TS has been proposed also for conservation issues as a tool to estimate biodiversity over large areas and in poorly known environments. This paper briefly reviews the use of TS in environmental impact studies and the effects of TS on sampling procedures and data analyses. The risk of possible loss of information depending on TS and the studied environment are discussed. Concluding remarks deal with the dangers of loss of taxonomic expertise in marine biological studies and assess critically the proposal of TS as a tool to describe biodiversity at a taxonomic level higher than species. PMID- 12735954 TI - Taxonomic level for assessing oil spill effects on soft-bottom sublittoral benthic communities. AB - Infralittoral soft-bottom macrofauna abundance data collected during four-year surveys after the Aegean Sea (Galicia, Spain) and the Amoco Cadiz oil spill (Brittany, France) at a total of seven sites were analysed at three different taxonomic levels (species, genus, and family). The effects of the spills on macrofauna populations were very similar in the two areas. The post-spill time courses of Shannon-Weaver diversity values were similar regardless of whether species-, genus- or family-level data were used. Non-metric multidimensional scaling identified well-defined groups of samples corresponding to different times post-spill, and again the results obtained were very similar regardless of the taxonomic level considered. In general, the use of genus- or family-level data did not lead to relevant information loss by comparison with species-level data. In view of these findings and those of previous studies of this type, we conclude that family-level data is generally sufficient for monitoring pollution effects in infralittoral soft-bottom environments. PMID- 12735955 TI - Changes in zooxanthellae density, morphology, and mitotic index in hermatypic corals and anemones exposed to cyanide. AB - Sodium cyanide (NaCN) is widely used for the capture of reef fish throughout Southeast Asia and causes extensive fish mortality, but the effect of NaCN on reef corals remains debated. To document the impact of cyanide exposure on corals, the species Acropora millepora, Goniopora sp., Favites abdita, Trachyphyllia geoffrio, Plerogyra sp., Heliofungia actinformis, Euphyllia divisa, and Scarophyton sp., and the sea anemone Aiptasia pallida were exposed to varying concentrations of cyanide for varying time periods. Corals were exposed to 50, 100, 300, and 600 mg/l of cyanide ion (CN(-)) for 1-2 min (in seawater, the CN(-) forms hydrocyanic acid). These concentrations are much lower than those reportedly used by fish collectors. Exposed corals and anemones immediately retracted their tentacles and mesenterial filaments, and discharged copious amounts of mucus containing zooxanthellae. Gel electrophoreses techniques found changes in protein expression in both zooxanthellae and host tissue. Corals and anemones exposed to cyanide showed an immediate increase in mitotic cell division of their zooxenthellae, and a decrease in zooxanthellae density. In contrast, zooxanthellae cell division and density remained constant in controls. Histopathological changes included gastrodermal disruption, mesogleal degradation, and increased mucus in coral tissues. Zooxanthellae showed pigment loss, swelling, and deformation. Mortality occurred at all exposure levels. Exposed specimens experienced an increase in the ratio of gram-negative to gram positive bacteria on the coral surface. The results demonstrate that exposure cyanide causes mortality to corals and anemones, even when applied at lower levels than that used by fish collectors. Even brief exposure to cyanide caused slow-acting and long-term damage to corals and their zooxanthellae. PMID- 12735956 TI - A database of archived drilling records of the drill cuttings piles at the North West Hutton oil platform. AB - Drill cuttings piles are found underneath several hundred oil platforms in the North Sea, and are contaminated with hydrocarbons and chemical products. This study characterised the environmental risk posed by the cuttings pile at the North West Hutton (NWH) oil platform. Data on the drilling fluids and chemical products used over the platform's drilling history were transferred from archived well reports into a custom database, to which were added toxicological and safety data. Although the database contained many gaps, it established that only seven chemical products used at NWH were not in the lowest category of the Offshore Chemicals Notification Scheme, and were used in only small quantities. The study therefore supports the view that the main environmental risk posed by cuttings piles comes from hydrocarbon contamination. The (dated) well records could help future core sampling to be targeted at specific locations in the cuttings piles. Data from many platforms could also be pooled to determine generic 'discharge profiles.' Future study would benefit from the existence, in the public domain, of a standardised, 'legacy' database of chemical products. PMID- 12735957 TI - Organochlorine contaminant exposure and associations with hematological and humoral immune functional assays with dam age as a factor in free-ranging northern fur seal pups (Callorhinus ursinus). AB - Potential effects of organochlorine contaminant (OC) exposure on humoral immune function and health of free-ranging northern fur seals were investigated. Forty two "neonates" were captured for blood sample collection and re-sampled as "pups" 29-51 days later. OCs were extracted from whole blood to identify polychlorinated biphenyl congeners and chlorinated pesticides by high performance liquid chromatography. Humoral immune function was assessed by antibody responses to vaccination and immunoglobulin concentrations. Additional health status indicators included leukocyte counts and haptoglobin concentrations. Mean OC concentrations were higher in neonates than at recapture. Neonates of young dams had higher mean OC concentrations than neonates of older dams. A higher proportion of old dam's pups developed a twofold or greater increase in antigen specific antibodies compared to young dam's pups. Higher OC exposure and poor immune responses in first-born pups may indicate a higher risk of secondary morbidity and mortality than pups of multiparous dams. PMID- 12735959 TI - Polycyclic aromatic hydrocarbons in the sediments of the Yalujiang Estuary, North China. AB - Selected polycyclic aromatic hydrocarbons (PAHs) in surface sediments of the Yalujiang Estuary, North China, have been investigated by gas chromatography-mass spectrometry (GC-MS) for flood season (August, 1994) and dry season (May, 1996), respectively. PAHs concentrations from sediments range from 68 to 1500 ngg(-1) depending upon the sample locations. The most contaminated sediment samples are found in the turbidity maximum area for both dry and flood seasons. The source of PAHs is most likely pyrolytic, with minor contributions from petrogenic and diagenetic PAHs for some samples. Perylene is mainly derived from biogenic inputs, and occurs at almost stations. In the Yalujiang Estuary, both biogenic and anthropogenic hydrocarbons are primarily derived form riverine discharges and are accumulated in the mixing zone. The overall levels of PAHs in this study are low compared to other regions and reveal moderate PAHs pollution in the Yalujiang Estuary. PMID- 12735958 TI - Removal of ammonia toxicity in marine sediment TIEs: a comparison of Ulva lactuca, zeolite and aeration methods. AB - Toxicity Identification Evaluations (TIEs) can be used to determine the specific toxicant(s), including ammonia, causing toxicity observed in marine sediments. Two primary TIE manipulations are available for characterizing and identifying ammonia in marine sediments: Ulva lactuca addition and zeolite addition. In this study, we compared the efficacy of these methods to (1) remove NH(x) and NH(3) from overlying and interstitial waters and (2) reduce toxicity to the amphipod Ampelisca abdita and mysid Americamysis bahia using both spiked and environmentally contaminated sediments. The utility of aeration for removing NH(x) and NH(3) during a marine sediment TIE was also evaluated preliminarily. In general, the U. lactuca and zeolite addition methods performed similarly well at removing spiked NH(x) and NH(3) from overlying and interstitial waters compared to an unmanipulated sediment. Toxicity to the amphipod was reduced approximately the same by both methods. However, toxicity to the mysid was most effectively reduced by the U. lactuca addition indicating this method functions best with epibenthic species exposed to ammonia in the water column. Aeration removed NH(x) and NH(3) from seawater when the pH was adjusted to 10; however, very little ammonia was removed at ambient pHs ( approximately 8.0). This comparison demonstrates both U. lactuca and zeolite addition methods are effective TIE tools for reducing the concentrations and toxicity of ammonia in whole sediment toxicity tests. PMID- 12735960 TI - Kinetic partitioning of Co, Mn, Cs, Fe, Ag, Zn and Cd in fresh waters (Loire) mixed with brackish waters (Loire estuary): experimental and modelling approaches. AB - To simulate the behavior of radionuclides along a salinity gradient, in vitro sorption and desorption kinetics of Co, Mn, Cs, Fe, Ag, Zn and Cd were studied in Loire river water and the macrotidal Loire estuarine water over two different seasons. Partitioning between the dissolved phase and suspended solids were followed up over 100 h after adding radioactive tracers to freshly collected freshwater (sorption stage); this stage was followed by desorption in fresh and estuarine waters. A kinetic model describing the interactions between trace metals and particles under a salinity gradient was developed and calibrated. Among parameters and/or processes that control the fate and behavior of contaminated particles during their transfer in estuarine systems, this study shows that the speciation of trace metals is controlled by: (i) the chemical water composition: for all the elements except for Fe, desorption increased with salinity; however, the amplitude of such an effect strongly depended on the element and/or on the composition of the particulate phase (and consequently on the season); (ii) the possibility for a given element to form (or not) stable surface particle moieties such as oxides or inner-sphere complexes; (iii) the distribution of a given element among different types of sites characterised by different binding forces that can lead (or not) to re-adsorption processes after mixing of contaminated particles with uncontaminated water. Our model enabled the quantification of the contribution and the characteristic time of reactions that took place over short and long periods on the global partitioning between particulate and dissolved phases during sorption and desorption and to determine the extent to which these reactions were modified by the salinity. PMID- 12735961 TI - Coral recruitment onto an experimental pulverised fuel ash-concrete artificial reef. AB - An experimental artificial reef was deployed in December 1993 at Hoi Ha Wan Marine Park, Hong Kong. This is the first study documenting natural scleractinian coral recruitment onto a stabilised pulverised fuel ash (PFA)-concrete artificial reef. Visible recruits were first recorded 9-10 months after the placement of reef blocks, i.e., in the autumn of 1994. Two scleractinians, Oulastrea crispata and Culicia japonica, were recruited. The recruit density of the former was much greater than the latter. The spatial recruitment pattern of the corals was observed to be affected by the orientation of the attaching surface. O. crispata settled predominantly on the undersides of the reef blocks. There was an edge effect on O. crispata recruitment. C. japonica, however, had a preference for exposed surfaces. O. crispata did not show a preference for block composition whereas C. japonica favoured blocks with high (75% by volume) PFA levels. This shows that PFA-concrete is a potential substratum for artificial reef construction, especially when such reefs aim at rehabilitating corals. PMID- 12735962 TI - Cryostorage of hepatic microsomes from two marine mammal species: effects on cytochrome P450-monooxygenase activities and content. PMID- 12735963 TI - Seasonal variation of selected metals in sediments, water and tissues of the groovy mullet, Liza dumerelii (Mugilidae) from the Mhlathuze Estuary, South Africa. PMID- 12735964 TI - Cloning of the HSP70 gene in barnacle larvae and its expression under hypoxic conditions. PMID- 12735965 TI - Chlorinated organic contaminants in surface sediments from the Yangtze Estuary and nearby coastal areas, China. PMID- 12735968 TI - Plutonium disproportionation: the ambiguity phenomenon. AB - Plutonium oxidation-state studies may yield ambiguous results if the parameters are not carefully chosen. The effect can be related to environmental plutonium as illustrated by an example. PMID- 12735969 TI - Theoretical and experimental determination of dosimetric characteristics for brachyseed Pd-103, model Pd-1, source. AB - Dosimetric characteristics of the BrachySeed Pd-103, Model Pd-1 source have been determined using both theoretical and experimental methods. Dose rate constant, radial dose function, and anisotropy functions of the source have been obtained following the TG-43 recommendations. Derivation of the dose rate constant was based on recent NIST WAFAC calibration performed in accordance with their 1999 Standard. Measurements were performed in Solid Water using LiF TLD chips. Theoretical simulation calculations were performed in both Solid Water and water phantom materials using MCNP4C2 Monte Carlo code using DLC-200 interaction data. The results of the Monte Carlo simulation indicated a dose rate constant of 0.65 cGyh(-1)U(-1) and 0.61 cGyh (-1)U(-1) in water and Solid Water, respectively. The measured dose rate constant in Solid Water was found to be 0.63+/-7% cGyh (-1)U( 1), which is within the experimental uncertainty of the Monte-Carlo simulated results. The anisotropy functions of the source were calculated in both water and in Solid Water at the radial distances of 1 to 7 cm. Measurements were made in Solid Water at distances of 2, 3, 5, and 7 cm. The Monte-Carlo calculated anisotropy constant of the new source was found to be 0.98 in water. The tabulated data and 5th order polynomial fit coefficients for the radial dose function along with the dose rate constant and anisotropy functions are provided to support clinical use of this source. PMID- 12735970 TI - Syntheses, in vitro and in vivo characterization of a 99mTc-(I)-tricarbonyl benzylamino-dihydroxymethyl phosphine (NP(2)) chelate. AB - Studies were performed to study the complexation chemistry of 99mTc(CO)(+)(3) with a new tridentate amino-dihydroxymethyl phosphine (NP(2)) ligand with the 99mTc(CO)(3)(OH(2))(+)(3) synthon at tracer levels. A single, well-defined 99mTc(CO)(3)NP(2) complex is formed at pH 7.5 within 10 min at 60 degrees C that exhibits high in vitro and in vivo stability. PMID- 12735971 TI - Preparation of (188) Re-labeled paper for treating skin cancer. AB - For homogeneous delivery of beta radiation to skin cancer, we developed a simple method for preparing (188) Re-labeled nitrocellulose paper. The homogeneity and stability of the labeled paper were investigated. Absorbed dose estimates were calculated using the Monte-Carlo method. A 74-MBq (188) Re-labeled paper with 1 cm diameter delivered 147.2 Gy up to 1-mm depth after 2-h irradiation. Animal experiments on tumor-bearing mice showed that 50 Gy is an adequate dose for treating skin cancer. Tumors disappeared 7 days after irradiation in all the groups irradiated by 50 or 100 Gy. The (188) Re-labeled paper provided a convenient, economical, effective, and non-invasive method of treating skin cancer. PMID- 12735972 TI - Improved synthesis of pure [18F]fluoro-compounds for PET studies from bromo compounds. AB - For preparing [18F]labeled compounds free from bromo-compounds for PET (positron emission tomography) studies, we propose the following two processes in which no separation is needed between the fluoro-compound and bromo-compound : (1). the bromo-compound is first converted into O-tosylate, which is then [18F]fluorinated; and (2). after [18F]fluorination of the bromo-compound, its excess is converted into an easily separable compound, e.g., phthalimide compound. Direct tosylation with silver tosylate was effected in 65-85% yield for most commercial bromo-compounds, and with potassium phthalimide, the bromo compounds in cold run were converted easily into phthalimide-compounds in 84-90% yield. PMID- 12735974 TI - Efficient synthesis of the 18F-labelled 3-O-methyl-6-[18F]fluoro-L-DOPA. AB - The 18F-labelled amino acid derivative 3-O-methyl-6-[18F]fluoro-L-DOPA ([18F]OMFD) is a potential radiotracer for imaging tumour tissue using positron emission tomography. The precursor N-formyl-3-O-methyl-4-O-boc-6-trimethyl stannyl-L-DOPA-ethyl ester enables direct electrophilic radiofluorination by stereoselective destannylation. After partial hydrolysis, optimized HPLC purification and sterile filtration, the [18F]OMFD obtained with high radiochemical purity and ready for use. The total synthesis time is about 50 min and the radiochemical yield achieved is 20-25% (decay corrected, related to [18F]F(2)). It was demonstrated that [18F]OMFD could be produced routinely and reliably for clinical use. [18F]FDOPA-preparation devices can be used with minor modifications. PMID- 12735973 TI - 90Y-oxine-ethiodol, a potential radiopharmaceutical for the treatment of liver cancer. AB - Ethiodol (or lipiodol) is selectively retained in hepatocellular carcinoma and is used as a vehicle to deliver radioactive agents following intraarterial hepatic infusion. We prepared the lipophilic complex 90Y-oxine with a radiolabeling efficiency of 97.6+/-1.1%. After extraction into ethiodol, a stability test in serum at 37 degrees C showed that 87.8% of the 90Y remained ethiodol-bound for 7 days. Bremsstrahlung imaging of a rabbit for 48 h confirmed that the homogeneous mixture of radiolabeled 90Y-oxine and ethiodol stayed in the targeted liver lobe. This radiopharmaceutical is thus a potential candidate for the treatment of non resectable liver cancer. PMID- 12735975 TI - An anticoincidence-shielded gamma-ray spectrometer for analysis of low level environmental radionuclides. AB - We developed an ultralow-level background gamma-ray spectrometer, using active and passive shield devices at the same time. Cosmic-ray-induced background is suppressed by means of active shield devices consisting of plastic scintillating plates of 50mm thick and anti-coincidence electronic system. The observed background rate was 0.34 s(-1) (=0.12s(-1) per 100 cm(3) Ge volume) for energy regions between 50 and 3000 ke V. The detection efficiency curve for 10(3)ml Marinelli beaker samples is obtained over all the energy regions. The advantages of the method are demonstrated by measuring the activity of 137Cs in powdered milk sample prepared without taking any chemical procedure. The MDA for 137Cs is estimated to be (17+/-1.7)mBq at a confidence level of 95% and it is about a factor of 10 lower than the MDA obtained from the previous cryostat assembly with 10-cm thick lead shielding. PMID- 12735976 TI - A review of the triple-to-double coincidence ratio (TDCR) method for standardizing radionuclides. AB - This paper gives a review of the development of the triple-to-double coincidence ratio (TDCR) method for standardizing pure beta- and pure electron capture (EC) radionuclides. The initial idea of Schwerdtel and practical realization and elaboration of the TDCR method by Pochwalski is presented. The main aspects and theoretical principle of the method are summarized and progress on the development of the theoretical model is presented. The details of the activity determination of pure beta- and pure EC radionuclides are discussed, along with specific details in the application of the method in each case. The enhanced TDCR method based on the MAC3 electronic system developed in France is also discussed. Different applications of the TDCR method in metrology laboratories around the world are also pointed out. PMID- 12735977 TI - Performance of a pencil ionization chamber in various radiation beams. AB - Pencil ionization chambers were recommended for use exclusively in the computed tomography (CT) dosimetry, and, from the start, they were developed only with this application in view. In this work, we studied the behavior of a pencil ionization chamber in various radiation beams with the objective of extending its application. Stability tests were performed, and calibration coefficients were obtained for several standard radiation qualities of the therapeutical and diagnostic levels. The results show that the pencil ionization chamber can be used in several radiation beams other than those used in CT. PMID- 12735979 TI - Determination of half-life and photon emission probabilities of (65)Zn. AB - A (65) Zn radioactive solution standardized by an absolute measurement method at Laboratoire National Henri Becquerel (LNHB), was sent to Bureau International des Poids et Mesures (BIPM), in order to include the results in their database. The activity value determined at BIPM was in good agreement with that stated by LNHB (the difference was 0.3%). Additional measurements of (65) Zn activity and half life were also carried out, by using a 4pigamma ionization chamber; the half-life value proposed by this work: 244.15 (10) days, is consistent to others reported in the literature. The spectra analysis was performed by gamma-ray spectrometry with high-purity germanium detectors, with the aim to improve the accuracy of the photon emission probabilities values of this nuclide. The determined photon emission probabilities are 0.498 for the 1115 ke V gamma-ray, 0.347 and 0.0479 for the X-rays Cu-K(alpha) and Cu-K(beta); the associated uncertainties are 0.4%, 0.9% and 1.1%, respectively. PMID- 12735978 TI - An agreed statement on calculating lead concentration and uncertainty in XRF in vivo bone lead analysis. PMID- 12735980 TI - Interwell tracing by environmental isotopes at Fimkassar Oilfield, Pakistan. AB - Water flooding is an important operation to enhance oil recovery. Movement of injected water is traced to test the performance of the water flood and verify suspected geological barriers or flow channels, etc. This paper describes the application of environmental isotopes as interwell tracers at Fimkassar Oilfield in Chakwal District, Pakistan. Water flooding was started in March 1996 in the Sakessar formation (Eocene). When water appeared in the production well in June 1998, isotopes (18O, 2H and 3H) were used to determine the breakthrough/transit time of the water flood and the contribution of fresh injected water. Water samples were collected during June 1998-August 1999 from the injection well, the production well, and some other fields for reference indices of the Sakessar formation and analyzed for the 18O, 2H and 3H, and chloride contents. The isotopic results confirmed the breakthrough of injected water and indicated that the water in the production well was a mixture of the injected water and the formation water. The initial breakthrough time was 27 months. The contribution of injected water varied from 67% to 80%. This percentage did not change significantly from the time of breakthrough until the last sampling indicating good mixing in the reservoir and the absence of any high permeability channel. Chloride content did not give any information about the hydraulic connection between the injection well and the production well. The study proved the potential of environmental isotopes for interwell tracing. PMID- 12735983 TI - Benzoyl nitrogen mustard derivatives of benzoheterocyclic analogues of netropsin: synthesis and biological activity. AB - Synthesis, DNA binding properties and biological activity of a series of bis benzoheterocycle derivatives 5-11, structurally related to the natural dipyrrole antitumor agent netropsin, and tethered to a benzoyl nitrogen mustard (BAM) as alkylating moiety is reported and structure-activity relationships determined. These compounds 5-11 have been evaluated for sequence selective alkylating properties and cytotoxicity against murine L1210 and human K562 leukaemia cells. Using as target sequence a portion of the long terminal repeat of the type-1 human immunodeficiency virus, we found that these compounds induce similar patterns of DNA fragmentation. In addition, the results obtained indicate that all synthesized compounds retain a good antiproliferative activity in the submicromolar range, and generally are more active against L1210 than K562 cells. With respect to both these cell lines, compounds 6, 7, 10 and 11 showed the greatest potency, ranging from 0.3 to 1 microM, while compounds 8 and 9 exhibit the lowest activity (IC(50)=2-12 microM). Among compounds 5-11, the derivative 11 was found to be the most potent member of this class and it is 5 and 10-fold less active than the bis-pyrrole counterpart 2 against K562 and L1210 cell lines, respectively. For compound 11, the substitution of the C-terminus benzofurane with N-methylindole and indole (to give the compounds 5 and 6, respectively) led to a decrease in cytotoxicity, which is more evident against the K562 cell line. Finally, differences were found among compounds 5-11 in induction of K562 differentiation. Some of them (compounds 7, 8 and 9) are potent inducers of erythroid differentiation of K562 cells, and could be proposed for differentiation anti-cancer therapy. PMID- 12735982 TI - Pyrrolo[2,1-d][1,2,3,5]tetrazine-4(3H)-ones, a new class of azolotetrazines with potent antitumor activity. AB - Pyrrolo[2,1-d][1,2,3,5]tetrazinones 10a-o, compounds that hold the deaza skeleton of the antitumor drug temozolomide, were prepared by reaction of 2-diazopyrroles 9 and isocyanates. Such a synthetic route represents, among those leading to azolotetrazinones reported so far, the only possible one since attempts to cyclize to the title ring system 2-amino-1-carbamoylpyrroles 11 or the mono substituted 2-triazenopyrrole 12 failed. Compounds 10 were screened at the National Cancer Institute (NCI) for their activity against a panel of about 60 human tumor cell lines. Most of them possess remarkable antineoplastic activity having GI(50) values in the low micromolar or sub-micromolar range and reaching, in the case of compound 10d, nanomolar concentrations. The most sensitive cell lines were MDA-N and MDA-MB-435 of the breast sub-panel, and SR, K-562, HL60 (TB) and CCRF-CEM of the leukaemia sub-panel. SAR evaluation and COMPARE computations indicate, for compounds 10, a mechanism of action different from that of temozolomide. PMID- 12735984 TI - Synthesis of 9-oxime-11,12-carbamate ketolides through a novel N-deamination reaction of 11,12-hydrazonocarbamate ketolide. AB - A series of 9-oxime-11,12-carbamate ketolides was synthesized for the first time through a key 11,12-hydrazonocarbamate intermediate that was first oximated and further deaminated to give the corresponding carbamate. The N-N bond cleavage was achieved through an original new reaction using glycoaldehyde dimer as deaminating reagent. The new compounds synthesized were shown to display improved antibacterial activities against Streptococcus pneumoniae and S. pyogenes resistant to erythromycin. PMID- 12735985 TI - Benzothiadiazine dioxides (BTD) derivatives as non-nucleoside human cytomegalovirus (HCMV) inhibitors. study of structural requirements for biological activity. AB - Two new series of BTD derivatives have been synthesised allowing to explore the steric requirements for their biological activity. The N3-alkylBTD compounds have shown antiviral activity in the same order or lower than previously prepared compounds. However, the cytotoxicity values observed prevent this new series of BTD derivatives from its potential therapeutic application. Concerning BTD derivatives with the modified linker attached to N1 position, we have obtained new non-nucleoside anti-HCMV derivatives. The activity against HCMV is shown at concentrations that were 10-fold lower than the concentration that was toxic for the host cells, which confirm that these derivatives show a specific antiviral effect against HCMV. SAR conclusions derived from these last compounds have provided new knowledge about the structural requirements of BTD showing certain positions that could be modified for enhancing the anti-HCMV action. PMID- 12735986 TI - Syntheses and biological evaluation of novel quinuclidine derivatives as squalene synthase inhibitors. AB - Squalene synthase (E.C. 2.5.1.21) catalyses the reductive dimerization of two molecules of farnesyl diphosphate to form squalene and is involved in the first committed step in cholesterol biosynthesis. Inhibition of this enzyme is therefore an attractive target for hypocholesterolemic strategies. A series of quinuclidine derivatives incorporating a tricyclic system was synthesized and evaluated for their ability to inhibit squalene synthase in vitro. A 9H-fluorene moiety was found to be optimal as the tricyclic system for potent inhibitory activity. Improved activity can be achieved with a conformationally constrained three-atom linkage connecting the tricyclic system with the quinuclidine nucleus. Among these compounds, (Z)-3-[2-(9H-fluoren-2-yloxy)ethylidene]-quinuclidine hydrochloride 31 was found to be a potent inhibitor of squalene synthase derived from hamster liver and human hepatoma cells with IC(50) values of 76 and 48 nM, respectively. Oral dosing of compound 31 demonstrated effective reduction of plasma non-HDL cholesterol levels in hamsters. PMID- 12735987 TI - Synthesis and activities of oxidative metabolites of the anti-arthritic drug candidate S-2474. AB - We have synthesized and characterized some oxidative metabolites of S-2474. In this study, we discovered a novel skeleton, the 2,3-dihydrobenzofuran derivative, which inhibited PGE(2) production at a very low concentration and was effective in the anti-carrageenin footpad edema assay. PMID- 12735988 TI - Origin of the stereospecificity in binding hydroxamates of alpha- and beta phenylalanine methylamide to thermolysin revealed by the X-ray crystallographic study. AB - Optically active N-formyl-N-hydroxy-alpha-phenylalanine methylamide (1) and N formyl-N-hydroxy-beta-phenylalanine methylamide (2) were evaluated as inhibitors for thermolysin (TLN) to find that while the D-form is more potent than its enantiomer in the case of the hydroxamate of alpha-Phe-NHMe, in the inhibition with hydroxamate of beta-Phe-NHMe, the L-isomer (K(i)=1.66+/-0.05 microM) is more effective than its enantiomer. In order to shed light on the stereochemical preference observed in the inhibitions, X-ray crystallographic analyses of the crystalline TLN.D-1 and TLN.L-2 complexes were performed to the resolution of 2.1A. While L-2 binds TLN like substrate does with its benzyl aromatic ring occupying the S(1)' pocket, the electron density in the S(1)' pocket in the complex of TLN.D-1 is weak and could best be accounted for by the methylcarbamoyl moiety. For both inhibitors, the hydroxamate moiety coordinates the active site zinc ion in a bidentate fashion. PMID- 12735989 TI - TAK-599, a novel N-phosphono type prodrug of anti-MRSA cephalosporin T-91825: synthesis, physicochemical and pharmacological properties. AB - Crystalline 1 (TAK-599) is a novel N-phosphono prodrug of anti-methicillin resistant Staphylococcus aureus (MRSA) cephalosporin 2a (T-91825) that has high affinity for penicillin-binding protein (PBP) 2' (IC(50); 0.90 microg/mL) and shows potent in vitro anti-MRSA activity (MIC against MRSA N133; 1.56 microg/mL), comparable to that of vancomycin (1.56 microg/mL). Although 2a had insufficient water solubility (2.3 mg/mL) for parenteral administration, 1 showed excellent water solubility (>100 mg/mL, pH 7) as well as good chemical stability in the solid state and solution. In pharmacokinetic studies, when 1 was administered intravenously to rats and monkeys, it was rapidly converted into 2a in the blood. These results show that 1 (TAK-599) is a highly promising parenteral cephalosporin targeted for MRSA infection. PMID- 12735990 TI - Synthesis and antihyperglycemic activity of suitably functionalized 3H-quinazolin 4-ones. AB - A series of 2-sec-amino-3H-quinazolin-4-ones (4a-p) and 4-sec-amino-2 chloroquinazolines (5a-b) have been synthesized by nucleophilic substitution reaction of 2-chloro-4(3H)-quinazolones (3) and 2,4-dichloroquinazolines (2) with amines, respectively. Most of the synthesized compounds were evaluated for antihyperglycemic activity but only 4a,b,d,j,o displayed significant reduction in blood glucose level in streptozotocin and sucrose loaded rat models. PMID- 12735991 TI - Site-specific incorporation of the 1-hexanol-1,N6-etheno-2'-deoxyadenosine adduct into oligodeoxyribonucleotides. AB - Modified oligonucleotides that contain the hydrophobic 1-hexanol-1,N(6)-etheno-2' deoxyadenosine adduct have been synthesized using a mild solid phase phosphoramidite chemistry. The presence and the integrity of the modified nucleoside in the synthetic oligomers were confirmed by electrospray ionization and MALDI mass spectrometry measurements together with analysis of the complete enzymatic hydrolysate by high performance liquid chromatography coupled to UV and fluorescent detection techniques. PMID- 12735992 TI - Synthesis of the cyclic and acyclic acetal derivatives of 1-(3-C-ethynyl-beta-D ribo-pentofuranosyl)cytosine, a potent antitumor nucleoside. Design of prodrugs to be selectively activated in tumor tissues via the bio-reduction-hydrolysis mechanism. AB - We have designed and synthesized the acetal derivatives of 1-(3-C-ethynyl-beta-D ribo-pentofuranosyl)cytosine (ECyd, 1), the 2',3'-O-nitrobenzylidene derivatives 2 and 3 and the 5'-O-(alkoxy)(nitrophenyl)methyl derivatives 6-10 as potential prodrugs of ECyd. These prodrugs can be selectively activated in tumor tissues via a bio-reduction-hydrolysis mechanism owing to the characteristic properties of tumor tissues, such as hypoxia and lower pH. Although the 2',3'-O-(4 nitrobenzylidene) derivatives 2 and 3 were converted bio-reductively into the corresponding 4-aminobenzylidene derivatives by rat S-9 mix, the reduction products, that is, the corresponding amino congeners 4 and 5, proved to be rather stable in an aqueous solution at pH 6.5 used as a pH model for acidic tumor tissues. In contrast, the 5'-O-(alkoxy)(4-nitropheny)methyl derivatives 6-8 were also reduced by rat S-9 mix to the corresponding amino congeners 11-13, which were hydrolyzed to release ECyd more effectively at pH 6.5 than at pH 7.4. Accordingly, the acyclic acetals 6-8 may be efficient prodrugs of ECyd, that are effectively reduced under physiological conditions releasing ECyd in acidic tumor tissues. PMID- 12735993 TI - Structural studies on McN-5652-X, a high-affinity ligand for the serotonin transporter in mammalian brain. AB - McN-5652-X, (+)-1, is a potent, selective inhibitor of serotonin reuptake in mammalian brain. When radiolabeled with (11)C, it has served as a positron emission tomography (PET) radiotracer for in vivo imaging of central serotonin transporters. We have determined the solid-state structures of (+)-1.HClO(4) and (+)-1.(+)-(2R,3R)-tartrate by single-crystal X-ray diffraction, thereby confirming the trans relative configuration (Chemical Abstracts Service rules of nomenclature) and the 6S,10bR absolute configuration for (+)-1. Results from a vibrational circular dichroism (VCD) study on (+)-1 in CDCl(3) are consistent with the 6S,10bR absolute configuration. PMID- 12735994 TI - New aspects of formation of 1,2-cyclic phosphates by phospholipase C-delta1. AB - Phosphoinositide-specific phospholipase C-delta1 (PI-PLC-delta1) cleaves phosphatidylinositol 4,5-bisphosphate (PI-4,5-P(2), 1), 5-phosphate (PI-5-P, 2) and 4-phosphate (PI-4-P, 3) to form the mixture of the corresponding 4,5-, 5- and 4-phosphorylated inositol 1,2-cyclic phosphate (IcP) and 1-phosphate (IP) (4-6 and 7-9, respectively). In this work, we have studied the rates of the cleavage and the ratios of the cyclic-to-acyclic phosphate products under various pH and Ca(2+) concentration conditions using 31P NMR to monitor the reactions. In agreement with the previous report (Kim et al. Biochim. Biophys. Acta 1989, 163, 177), our results indicate that the IcP/IP ratios strongly depend on the reaction conditions, with the cyclic phosphate products formed predominantly at low pH (pH 5.0) and high calcium concentration (5 mM). Surprisingly, however, we have found that at pH 8.0 and 5 mM Ca(2+), PI-5-P rather than PI-4,5-P(2) is the most preferred substrate with the highest V(max). The cleavage of PI-5-P generated also more cyclic phosphate product than the other two substrates. In addition, we have studied the analogous reaction of phosphorothioate analogues of 1 with the sulfur placed in the nonbridging (10) or bridging (13) positions. We have found that the phosphorothioate analogue 10 produced exclusively the cyclic product 11, whereas the analogue 13 afforded exlusively the acyclic product 7. These results are discussed in terms of the mechanism of PI-PLC, where the cyclic product is formed by 'leaking' from the active site before its subsequent hydrolysis. The potential significance of the cyclic products in the signaling pathways is also discussed. PMID- 12735995 TI - Synthesis and activity of N-benzyl pseudopeptides HIV protease inhibitors. AB - A series of N-benzyl pseudopeptides was designed, synthesized and tested as HIV-1 protease inhibitors. The ability of the new compounds containing N-benzyl hydroxyalkylamino acid core structure to inhibit HIV replication in cell culture is comparable to their capacity to inhibit the isolated enzyme, a result compatible with good pharmacokinetic properties of these derivatives. The pseudotripeptide Fmoc-Leu-N(Bzl)Hse-Met-NH-tBu was the best inhibitor of the series (IC(50)=170 nM) showing promising inhibition of viral replication (ED(50)=52 nM). All new compounds exhibit high enzymatic resistance and stability against cell cultures and plasma enzymes. PMID- 12735996 TI - Microelectromechanical systems in urology. PMID- 12735997 TI - Treatment responsiveness of the Self-Esteem And Relationship questionnaire in erectile dysfunction. AB - OBJECTIVES: To determine the treatment responsiveness of the disease-specific Self-Esteem And Relationship (SEAR) questionnaire in erectile dysfunction. METHODS: The SEAR questionnaire was administered at baseline and at the end of the study in 93 patients with erectile dysfunction enrolled in a 10-week, open label, flexible-dose (50-mg sildenafil, adjustable to 25 mg or 100 mg) trial. Changes from the baseline score were analyzed using the paired t test. The correlation between the changes from baseline on the SEAR questionnaire and the Erectile Function domain of the International Index of Erectile Function was examined. RESULTS: Significant and meaningful differences (P = 0.0001) from baseline were observed in the two primary domains (Sexual Relationship and Confidence) and the two Confidence domain subscales (Self-Esteem and Overall Relationship). The magnitude of the change was quite high for most aspects (Sexual Relationship, effect size [ES] = 1.6; Confidence, ES = 1.0; Self-Esteem, ES = 1.1) and moderate for one (Overall Relationship, ES = 0.6). Changes in Erectile Function domain score correlated moderately with changes in the SEAR domain and subscale scores (Sexual Relationship, r = 0.69; Confidence, r = 0.48; Self-Esteem, r = 0.47; and Overall Relationship, r = 0.35; P Brij 35>SDS>SDSo>LSB. The presence of cosolvents, such as Methanol, Dimethylformamide (DMFA) and 1,4 Dioxan (DX) at various volume percentages, increased the CMC of both SDS and Tween 20 and at a certain concentration totally inhibited the micellization. The binding of SO to micelles decreased as the concentration of the cosolvents increased. This inhibitory effect of cosolvents on binding of SO to micelles followed the order as; Methanol>DMFA>DX. PMID- 12736074 TI - Vibrational spectra of furan, pyrrole, and thiophene from a density functional theory anharmonic force field. AB - Quartic force fields for furan, pyrrole, and thiophene have been generated using density functional theory (DFT). These were used to evaluate vibrational levels by second-order perturbation theory (PT) and also by the variational method. The results for the fundamental frequencies are in very good agreement with observation. The accuracy of overtones and combination transitions is also good, for those cases where assignments can be made. Second-order PT combines speed and quality whereas the variational approach is inherently more reliable, but converges rather slowly, requiring significant computational effort. PMID- 12736076 TI - Fluorimetric estimation of DNA content using sensitized lanthanide fluorescence. AB - Lanthanide fluorescence enhancement on complexation with calf thymus DNA was studied in aqueous solution. The DNA sensitized and enhanced fluorescence of terbium and europium by nearly two orders of magnitude. By applying this ligand sensitized lanthanide fluorescence enhancement, DNA could be estimated at 10 ppb level. Further, effect of addition of TOPO in Triton X-100 micellar medium to Tb DNA complex in solution was also studied. On addition of TOPO, no synergistic terbium fluorescence enhancement was observed. PMID- 12736075 TI - Observation of CO(2) in Fourier transform infrared spectral measurements of living Acholeplasma laidlawii cells. AB - In monitoring the time course of conformational disorder by Fourier transform infrared spectroscopy for intact Acholeplasma laidlawii cells grown at 37 degrees C on binary fatty acid mixtures containing oleic acid and for cells grown on pure palmitic acid, an absorption band at 2343 cm(-1) was observed. The band intensity was found to increase with time. This band was not observed in the spectra for isolated membranes. It is suggested that the 2343 cm(-1) band is due to CO(2) dissolved in water, most likely produced at the final point of fermentation of amino acid by this microorganism. PMID- 12736077 TI - Electron paramagnetic resonance and ultraviolet/visible study of compounds I and II in the horseradish peroxidase-H(2)O(2)-silk fiber reaction system. AB - The enzymatic oxidation of silk with H(2)O(2) in the presence of horseradish peroxidase (HRP) has been investigated. Two intermediate complexes have been observed during this reaction. Both can be attributed to Fe(4+) ions axially bonded to an oxygen atom and to a porphyrin radical (P). In the most unstable of them, indicated as compound II, the chemical bond between [Fe(IV)=O](2+) and P was weaker than in the other, indicated as compound I. The former compound disappeared within 1 h of the reaction, at difference with the latter, traces of which were observed even after 3 weeks with dried samples. However, the chemical bond between [Fe(IV)=O](2+) and P in compound I weakened during the sample ageing. All these phenomena have been enlightened by electron paramagnetic resonance (EPR) and spectrophotometric ultraviolet/visible (UV/Vis) measurements. PMID- 12736078 TI - PET after cardiac arrest: does it have any role in resuscitation medicine research? PMID- 12736079 TI - Clarifying TCD criteria for brain death--are some arteries more equal than others? PMID- 12736080 TI - Unlike myofibers, neuromuscular junctions remain stable during prolonged muscle unloading. AB - This study assessed the effect of muscle unloading on the neuromuscular system. Sixteen male Fischer 344 rats were randomly assigned to either a hindlimb suspension (unloaded) or control group (N=8/group) for 16 days. Following this intervention period, pre- and postsynaptic features of the neuromuscular junctions (NMJs) of soleus muscles were stained with cytofluorescent techniques, and myofibers were histochemically stained for ATPase activity. The data indicate that 16 days of muscle unloading resulted in significant (P<0.05) atrophy among myofibers (>50%) that was evident among all three major fiber types (I, IIA and IIX), but failed to significantly alter any aspect of NMJ morphology quantified. These results demonstrate an impressive degree of NMJ resilience despite dramatic remodeling of associated myofibers. This may be of benefit during post-unloading rehabilitative measures where effective neuromuscular communication is essential. PMID- 12736082 TI - Etanercept (Enbrel) therapy for chronic inflammatory demyelinating polyneuropathy. AB - Patients with chronic inflammatory demyelinating polyneuropathy (CIDP) and/or variants who were refractory or intolerant of standard therapies were treated with etanercept, 25 mg twice per week. Ten patients underwent treatment, and manual muscle strength, sensory thresholds and functional abilities were tested prior to and 4-6 months after initiating therapy. Three patients had significant improvement and three others had possible improvement. Based on these preliminary observations, treatment with etanercept may be considered in patients with CIDP, who cannot undergo standard therapies, although its efficacy in CIDP needs to be examined in a double-blinded, controlled clinical trial. PMID- 12736081 TI - Scavenger receptor class B type I expression in murine brain and regulation by estrogen and dietary cholesterol. AB - The scavenger receptor class B type I (SR-BI), a receptor for high-density lipoproteins (HDL), facilitates cholesterol delivery to steroidogenic tissues, and brings excess body cholesterol to liver for excretion. Scavenger receptors are also involved in the internalization of aggregates of Alzheimer's disease (AD) amyloid beta-protein, and selective uptake of HDL-associated vitamin E in the brain. Therefore, modulation of the brain SR-BI may affect these processes. The present study examined the expression of SR-BI receptors in murine brain and their regulation by estradiol administration and cholesterol feeding. Liver and brain appeared to express similar SR-BI transcripts. Expression of SR-BI was highest in the adrenals and lowest in the brain. In rats, estradiol administration decreased SR-BI in liver, but increased in adrenals. In mice, estrogen treatment decreased hepatic SR-BI, but interestingly increased the levels of brain SR-BI mRNA. Cholesterol feeding did not alter mouse hepatic SR-BI mRNA, but increased brain SR-BI levels. ATP-binding cassette transporter A1 (ABCA1), involved in cellular cholesterol transport, increased in cholesterol-fed mouse liver, but did not show changes in the brain. These studies suggest that SR B1 is expressed in the brain and regulated by hormonal and nutritional stimuli, which may influence the pathophysiology of neurological disorders like AD. PMID- 12736083 TI - Cerebral perfusion and metabolism in resuscitated patients with severe post hypoxic encephalopathy. AB - Positron emission tomography (PET) was used for the study of regional cerebral perfusion and metabolism in eight patients with severe post-hypoxic encephalopathy, caused by cardiac arrest and resulting in a coma lasting for at least 24 h. Using this method, we aimed to identify regional vulnerability, which was hypothesized to provide (i) insight in pathogenic mechanisms and (ii) early prognostic parameters. On day 1 post-resuscitation, 18-Fluor deoxyglucose ([F18] FDG) indicated a marked decrease of cerebral metabolic activity. Gray matter glucose consumption was 54% of normal values, whereas white matter uptake was 70% of normal. Regional differences followed a pattern of neuronal density rather than specific patterns of functionally or biochemically defined regions or of vascular territories. In contrast to [F18]-FDG, the distribution of 15-oxygen labeled water ([O-15]-water) showed a better demarcation between gray and white matter, whereas focal deficit was not observed. In some patients, hyperperfusion relative to regional glucose consumption was observed in the occipital poles and basal ganglia. This suggests loss of vascular tone, i.e. vascular paralysis, in the basilar artery territory. CT and MRI scanning did not show any major change with respect to the hypoxic injury. In the small group studied, all patients had a poor outcome. The comparison between survivors and nonsurvivors did not reveal obvious differences in PET data, suggesting that this technique does not provide major prognostic clues adding to the prognostic information derived from serial neurological assessment in the restricted patient group characterized by prolonged coma. PMID- 12736085 TI - A novel insertion mutation in spastin gene is the cause of spastic paraplegia in a Chinese family. AB - A total of eight loci for autosomal dominant hereditary spastic paraplegia (ADHSP) has been mapped to chromosome 14q, 2p, 15q, 8q, 10q, 12q, 19q, 2q, respectively, among which the SPG4 gene on chromosome 2p21-22 encoding spastin, an ATPase of the AAA family, accounts for 40-50% of all ADHSP families and is expressed in both adult and fetal tissues. In this work, we reveal a novel insertion mutation in exon 11 of the SPG4 gene found in a big Chinese family composed of 47 members, including 20 affected ones, using linkage analysis. The mutation was well demonstrated to be the cause of loss of production of the functional protein by pre-termination of translation in AAA cassette region. To our knowledge, this is the first report of spastin mutation in China. PMID- 12736084 TI - Persistence of isolated flow in the internal carotid artery in brain death. AB - The criteria for the confirmation of brain death (BD) using transcranial Doppler ultrasonography (TCD) state that flow should not be observed in the internal carotid artery (ICA). In 94 brain-dead patients examined by TCD, 19 (20%) had flow in the ICA despite cerebral circulatory arrest. There was no difference in the clinical characteristics of patients with or without isolated flow in the ICA. Shunting of blood from the ICA into the external carotid system or the arrest of blood flow at a higher level than the carotid siphon may explain our findings. Our results show that the current TCD criteria can result in a high rate of false negatives. We suggest that the TCD criteria for the diagnosis of brain death should be reviewed. PMID- 12736086 TI - Anti-GM1b IgG antibody is associated with acute motor axonal neuropathy and Campylobacter jejuni infection. AB - Anti-GM1 and anti-GM1b antibodies are frequently present in patients with Guillain-Barre syndrome (GBS) and accordingly, the two antibodies often coexist in the same patient. In order to study clinical and laboratory features of anti GM1b-positive GBS, we analyzed the data of patients with anti-GM1b IgG antibody but no anti-GM1 IgG antibody. Of 86 consecutive patients, 10 had anti-GM1b antibody alone and frequently had acute motor axonal neuropathy (AMAN, 80%) and Campylobacter jejuni infection (60%). Of 10 patients with anti-GM1 antibody alone, four had AMAN, and two had C. jejuni infection. These results showed that GM1b could be a target molecule of autoantibody in the AMAN form of GBS subsequent to C. jejuni infection. PMID- 12736087 TI - Architectural abnormalities in muscle nuclei. Ultrastructural differences between X-linked and autosomal dominant forms of EDMD. AB - OBJECTIVES: The aim of our study was to compare the ultrastructure of myonuclei in both forms of Emery-Dreifuss dystrophy (EDMD)-X-linked and dominantly autosomally transmitted. The muscle biopsies were taken from rectus femoris in four X-linked EDMD cases and three ADEDMD cases. METHODS: The biopsies were evaluated using immunocytochemical staining to establish emerin or A/C lamins deficiency. The muscle ultrastructure, especially that of nuclei, was analysed to find out whether there are differences between the two forms of EDMD. RESULTS: In both forms of EDMD, there was an aberrant nuclear architecture. In the X-linked form, the breakdown of fragile nuclear membrane and presence of nucleoplasm extrusion were a distinct feature. In the AD from, there was chromatin reorganization and loss of nucleoplasm volume. PMID- 12736088 TI - D-cycloserine for the treatment of ataxia in spinocerebellar degeneration. AB - We studied the effects of D-cycloserine, a partial NMDA receptor allosteric agonist, on ataxia in patients with spinocerebellar degeneration. Fifteen Japanese ataxic patients enrolled in a 14-day single-blind trial of D-cycloserine (daily oral dose of 50 mg) following a 14-day single-blind placebo phase. At the end of the D-cycloserine administration, there was a significant reduction in the posture, gait and total score of the international cooperative ataxia rating scale and in the time for walking and speech tasks. D-Cycloserine was well tolerated and no adverse effect was observed. D-Cycloserine may have therapeutic efficacy for spinocerebellar ataxia. PMID- 12736089 TI - Study of the rostral midbrain atrophy in progressive supranuclear palsy. AB - Rostral midbrain atrophy in progressive supranuclear palsy (PSP) is detected by mid-sagittal plain magnetic resonance imaging (MRI). The shape of the atrophy looks like the bill of a hummingbird (hummingbird sign). We studied this sign to elucidate the nature of midbrain atrophy in PSP. Eight patients with PSP, 12 with Parkinson's disease (PD), and 10 normal controls were studied. Using mid-sagittal plain MRI, we measured the rostral and caudal midbrain tegmentum (MT), superior and inferior colliculus, pontine base, and tegmentum. We compared the length of the interpeduncular fossa, which is posterior to the mammillary body, to the diameter of the midbrain tegmentum. The multiple comparison method was used for the statistical analysis. The hummingbird sign was demonstrated in all of the PSP patients studied, and it was not observed in PD patients nor in normal controls. The hummingbird sign in the PSP patients was due to the atrophy of the midbrain tegmentum (rostral and caudal) and to a relative increase in the length of the interpeduncular fossa over that of the anteroposterior diameter of the midbrain tegmentum. The hummingbird sign, which represents the atrophy of the rostral midbrain tegmentum, strongly suggests the involvement of the rostral interstitial nucleus of the medial longitudinal fasciculus in patients with PSP. Demonstration of a hummingbird sign on MRI is thought to be useful for a diagnosis of PSP. PMID- 12736090 TI - Screening of the early growth response 2 gene in Japanese patients with Charcot Marie-Tooth disease type 1. AB - Charcot-Marie-Tooth disease type 1 (CMT1) is a heterogeneous disorder. Most CMT1 patients are associated with a duplication of 17p11.2-p12 (CMT1A duplication), but a small number of patients have mutations of peripheral myelin protein 22 (PMP22), myelin protein zero (MPZ), connexin 32 (Cx32) and early growth response 2 (EGR2) genes. In our previous study, we identified the responsible mutations in 72 of 128 Japanese CMT1 patients as CMT1A duplication in 40, PMP22 mutation in 6, MPZ mutation in 12 and Cx32 mutation in 14 patients. A total of 56 Japanese CMT1 patients with no identified mutations were screened for EGR2 mutation by denaturing gradient gel electrophoresis (DGGE). We detected a heterozygous Asp383Tyr mutation of EGR2 in one patient with severe CMT1, Dejerine-Sottas syndrome. EGR2 mutation is rare cause of CMT1 in Japan as in other nations. We were unable to identify the responsible mutation in 55 of 128 CMT1 patients and need further analysis to identify their candidate genes. PMID- 12736091 TI - A new measure of dysconjugacy in INO: the first-pass amplitude. AB - BACKGROUND: The ratios of abducting to adducting eye movements (versional dysconjugacy index, VDI) for saccadic velocity and acceleration have been useful measures by which to objectively characterize internuclear ophthalmoparesis (INO). Amplitude measures of dysconjugacy have been less useful, given that many patients maintain the ability to ultimately reach a centrifugal fixation target and that traditional amplitude measures of VDI have focused on this 'final amplitude' (FA) position. METHODS: We utilized infrared oculography to define a new amplitude measure of dysconjugacy in 42 multiple sclerosis (MS) patients with INO. The first-pass amplitude (FPA)-VDI is the ratio of abduction/adduction eye movement amplitudes computed at the time when the abducting eye initially achieves the centrifugal horizontal fixation target. RESULTS: FPA-VDI values were significantly more sensitive and specific than FA-VDI values in demonstrating dysconjugacy in INO, and there was a 14-fold increase in dysconjugacy as measured by FPA-VDI Z-scores when compared to FA-VDI Z-scores. CONCLUSION: Consideration of velocity (pulse) and amplitude (step) components of dysconjugacy in patients with INO can provide a greater understanding of the dynamic aspects of this syndrome. We propose to characterize the relationship between the pathophysiology of INO and neuroradiologic measures of tissue injury in MS. PMID- 12736092 TI - Sexual dysfunction in multiple sclerosis: a MRI, neurophysiological and urodynamic study. AB - We studied 31 patients with relapsing-remitting (RR) multiple sclerosis (MS) in which we performed an urodynamic study, the pudendal cortical evoked potentials, the tibial cortical evoked potentials and the cranial and cervical spinal cord magnetic resonance imaging (MRI). We calculated the T(1) and T(2) lesion load (LL) and brain parenchymal fraction (BPF) of whole brain, frontal lobes, pons and cervical spinal cord. We also estimated the cross-sectional area at C(2) level. Spearman's rank correlation analysis showed a relationship between symptoms of sexual dysfunction and age (r=0.73, p<0.0001), cognitive performances (r=-0.63, p<0.0001), level of independence (r=-0.63, p<0.0001), disability (r=0.56, p<0.001), symptoms of anxiety (r=0.55, p<0.001) and depression (r=0.50, p<0.005), disease duration (r=0.42, p<0.02) and parenchymal atrophy in the pons (r=-0.38, p=0.031). Sexual dysfunction was not correlated with any other MRI measure, urodynamic patterns or cortical evoked potentials. In multiple regression analysis, sexual dysfunction was predicted only by T(1) lesion load of the pons. In conclusion, we confirmed previous correlations of sexual dysfunction with various clinical variables and demonstrated an association between sexual dysfunction and destructive lesions in the pons, as detected by MRI, in patients with relapsing-remitting multiple sclerosis. PMID- 12736093 TI - Apolipoprotein E, angiotensin-converting enzyme and alpha-1-antichymotrypsin genotypes are not associated with post-stroke dementia. AB - There is evidence that indicates the involvement of environmental and genetic factors in the pathogenesis of post-stroke dementia (PSD). In the present work, we examined different polygenic influences on the risk of PSD in a series of stroke patients. We studied 150 consecutive patients evaluated 3 months after suffering acute strokes. All patients were evaluated with a prospective standard protocol and genotyped for vascular disease-associated polymorphisms in the genes coding for apolipoprotein E (including apoE coding and apoE promoter polymorphisms), angiotensin-converting enzyme (ACE) and alpha-1-antichymotrypsin (ACT). Thirty-two cases (21.3%) resulted in dementia 3 months after the stroke. In patients with PSD, the frequency of apoE epsilon 4 (0.08), ACE-D (0.64), ACT-A (0.62) alleles and apoE gene promoter polymorphisms (-491/A, 0.88; -427/C, 0.02) was similar to that of patients without PSD (apoE epsilon 4: 0.10, p=0.79; ACE-D: 0.56, p=0.36; ACT-A: 0.51, p=0.21; -491/A: 0.86, p=1.00; -427/C: 0.08, p=0.29). Our data indicate that PSD is not associated with the genetic risk factors of vascular dementia (VD) that were studied, and that additional factors may contribute to the pathogenesis of PSD. PMID- 12736094 TI - Motor responses evoked by transcranial magnetic stimulation and peripheral nerve stimulation in the ulnar innervation in amyotrophic lateral sclerosis: the effect of upper and lower motor neuron lesion. AB - We studied the upper (UMN) and lower motor neuron (LMN) innervations of 159 hands from 81 patients with amyotrophic lateral sclerosis (ALS). Eleven patients with various chronic LMN disorders causing weakness in the abductor digiti minimi (ADM) muscle served as LMN controls. Thirty healthy subjects served as normal controls. Cortical motor threshold, central conduction time (CMCT), and motor evoked response amplitude (MEP) after transcranial magnetic stimulation (TMS) were studied, and the MEP/M wave ratio was calculated. The data was analyzed in the ALS subjects in groups defined by ADM muscle strength and by the presence or absence of clinical signs of UMN involvement. CMCT was not increased in the ALS or LMN disease groups. The threshold was higher in limbs with both weak ADM muscles and UMN signs. The MEP/M wave amplitude ratio was increased in weak muscles in the ALS patients, notably in limbs with no UMN signs, and also in weak muscles in patients with other chronic LMN disorders. It was frequently decreased in strong muscles. There was no difference between bulbar-onset and limb-onset ALS groups, and there was no correlation between threshold and disease duration. We suggest that expressing the data as an index and utilising the MEP/M wave amplitude ratio as a variable is a sensitive method for detecting UMN abnormality in ALS in particular in early affected muscles. PMID- 12736096 TI - Transient global amnesia associated with an acute infarction in the retrosplenium of the corpus callosum. AB - We present a patient with transient global amnesia (TGA) whose diffusion-weighted imaging (DWI) study showed a high-intensity signal in the left retrosplenium of the corpus callosum. In previous studies, lesions in the retrosplenium caused permanent but not transient global amnesia, called retrosplenial amnesia, by involving the thalamocortical portion of the Papez' circuit. This is the first report indicating that TGA can be associated with acute infarction only in the left retrosplenium of the corpus callosum. PMID- 12736095 TI - A novel insertion mutation of acetazolamide-responsive episodic ataxia in a Japanese family. AB - We report a Japanese family with acetazolamide-responsive episodic ataxia. The proband was a 41-year-old woman with interictal nystagmus. She experienced recurrent attacks of loss of equilibrium and loss of coordination of the extremities accompanied by dysarthria and nausea beginning at about 10 years old. These episodes usually lasted for several hours two or three times a week. Direct sequence of CACNA1A demonstrated a novel insertion mutation in the patient and her father. This mutation is estimated to cause early stop of the gene transcription, producing a truncated protein. This is the first report of episodic ataxia type 2 of which the mutation was identified in a Japanese family. PMID- 12736097 TI - Longitudinal changes of anti-ganglioside antibodies before and after Guillain Barre syndrome onset subsequent to Campylobacter jejuni enteritis. AB - Anti-ganglioside antibodies frequently are present in sera from patients with Guillain-Barre syndrome (GBS) during the acute phase, but no patients in whom anti-ganglioside antibodies were tested before the onset of the syndrome have been reported. We describe the first case of GBS subsequent to Campylobacter jejuni infection, in which longitudinal changes in anti-ganglioside antibody titers were measured before and after the onset of limb weakness. Serum antibody titers against GM1 (IgM/IgG), GM1b (IgM/IgG), GalNAc-GD1a (IgM/IgG), and GD1b (IgG) were highest on the day of onset, but negative before onset. Anti-C. jejuni IgG and IgA antibody titers paralleled those of the anti-ganglioside antibodies, indicative that C. jejuni infection triggered anti-ganglioside antibody production. Press et al. [J. Neurol. Sci. 190 (2001) 41] reported that anti ganglioside antibody titers peaked during the recovery phase, but our findings are counter to theirs. We speculate that anti-ganglioside antibodies are the primary effectors of nerve damage in GBS. PMID- 12736098 TI - Dissociation between titer of anti-ganglioside antibody and severity of symptoms in a case of Guillain-Barre syndrome with treatment-related fluctuation. AB - Since plasma exchange (PE) and intravenous immunoglobulin (i.v.Ig) have been widely used in treatment for Guillain-Barre syndrome (GBS), early relapse and treatment-related fluctuation have been a potential problem, but little is known about the mechanism of relapse and fluctuation. We describe a patient who had GBS with treatment-related fluctuation. A 37-year-old Japanese man exhibited acute distal-dominant weakness in upper limbs after upper respiratory infection. His cranial nerve system was normal and muscle weakness was limited to upper limbs. Anti-GT1a IgG was strongly positive and anti-GQ1b IgG was also detected in his serum. Muscle weakness responded well to double-filtration plasmapheresis (DFPP) followed by i.v.Ig, but relapsed 45 days after the initial treatment. Although repeated treatments were effective, the patient showed additional minor deterioration twice. Motor nerve conduction velocities (MCVs) corresponded to the muscle weakness, but elevated level of cerebrospinal fluid (CSF) protein remained and anti-ganglioside antibody titers steadily decreased throughout the clinical course. These findings indicate that the clinical fluctuation was not due to changes in the production of anti-ganglioside antibodies but presumably to the transient beneficial effects of DFPP/i.v.Ig and the outlasting inflammatory response in peripheral nerves. PMID- 12736100 TI - Editorial comment on "Reduction in the number of sentinel lymph node procedures by preoperative ultrasonography of the axilla in breast cancer" by Deurloo and colleagues. PMID- 12736101 TI - Extended neoadjuvant chemotherapy in locally advanced breast cancer combined with GM-CSF: effect on tumour-draining lymph node dendritic cells by Pinedo et al. PMID- 12736102 TI - Molecular biomarkers of colorectal carcinogenesis and their role in surveillance and early intervention. AB - Modern medicine is increasingly focused towards population surveillance for disease, coupled with the implementation of preventative measures applied to 'at risk' patients. Surveillance in colorectal cancer is limited by the cost and risk of endoscopy. Trials of putative chemopreventive agents in colorectal cancer are hampered by difficulties in following up large cohorts of patients over long periods of time to ascertain the clinical effect. Research into possible pathways of colorectal carcinogenesis has revealed a range of biological intermediates which could be used in surveillance, the identification of high risk populations and early diagnosis of cancer. The aim of this paper was to review the possible role of biomarkers in surveillance and the timing of intervention. A literature review using both Medline and Web of Science was performed from 1995 onwards using keywords: biomarkers, colorectal cancer, carcinogenesis, chemoprevention, surveillance and screening. Research has identified many potential biomarkers, such as cyclooxygenase-2 (COX-2), oxidative DNA adducts and glutathione S transferase (GST) polymorphisms, which could be applied in a clinical setting to screen for and detect colorectal cancer. Molecular biomarkers, such as COX-2, oxidative DNA adducts and GST polymorphisms offer new prospects in the detection of early colorectal cancer, surveillance of high-risk populations and prediction of the clinical effectiveness of chemopreventive drugs. Their role could be extended into surgical surveillance for potentially operable disease and post operative follow-up for disease recurrence. Research should be directed at assessing complementary biomarkers to increase clinical effectiveness in determining management options for patients. PMID- 12736103 TI - Improving cancer therapy by non-genotoxic activation of p53. AB - Inactivation of p53 function is a common event in cancer. Approximately 50% of human tumours express mutant p53 and there is evidence that in others, including many childhood tumours, p53 function is impaired in other ways. These defects in p53 function may be due to the alteration of cellular factors that modulate p53 or to the expression of viral oncoproteins. Radiotherapy and many of the chemotherapeutic drugs currently used in cancer treatment are potent activators of p53. However, most of these therapies have a serious drawback, and that is the long-term consequences of their DNA damaging effects. Here, we review the discoveries in p53 research that are most significant to the development of new therapies based on the induction of the transcriptional activity of p53 in a non genotoxic way and discuss the situations in which this type of approach may be most beneficial. PMID- 12736104 TI - Extended neoadjuvant chemotherapy in locally advanced breast cancer combined with GM-CSF: effect on tumour-draining lymph node dendritic cells. AB - The effect of long-term administration of granulocyte-macrophage colony stimulating factor (GM-CSF) on dendritic cell (DC) activation and survival in patients with locally advanced breast cancer (LABC) was studied. To this end, the number of activated DC (i.e. positive for the marker S100) in tumour-draining lymph nodes (TDLN) was determined and compared between LABC patients receiving neoadjuvant chemotherapy with GM-CSF (n=52) or without GM-CSF (n=11), and a control group of chemonaive breast cancer patients (n=10). A significantly higher mean percentage of S100+ DC in the TDLN of the GM-CSF-treated patients (9.9%) was found compared with each of the respective control groups (5.3 and 5.1%, P=0.002). Moreover, intrapatient comparison before and after treatment showed that the percentage of S100+ DC significantly increased over the course of the GM CSF treatment (P=0.018). In a univariate survival analysis with a median follow up of 64 months, relatively high percentages of S100+ DC (> or =8%) were associated with a longer disease-free survival (DFS) (P=0.078). In patients with a high tumour load, where immunosuppressed conditions generally prevail, long term administration of GM-CSF may thus contribute to survival through enhanced DC activation and consequently improved chances of effective antitumour immunity. PMID- 12736105 TI - Reduction in the number of sentinel lymph node procedures by preoperative ultrasonography of the axilla in breast cancer. AB - Currently, breast cancer patients without clinically suspicious lymph nodes are candidates for sentinel lymph node procedures (SLNPs). The aims of this study were to investigate whether preoperative axillary ultrasonography and fine-needle aspiration cytology (FNA) can reduce the number of the more time-consuming SLNPs, and to identify a subset of quantitative nodal features to predict metastatic involvement. 268 axillae were ultrasonographically examined. FNA was performed on suspicious nodes (smallest diameter > or =5 mm or atypical cortex appearance). SLNP was omitted if a tumour-positive node was found on FNA. Length, width, maximum cortex thickness and appearance of cortex and hilus were ultrasonographically established. In 93 axillae (35%), at least one node was detected with ultrasound. FNA was performed once per axilla on 66 nodes; 37 (56%) contained tumour cells. 31% of all tumour-positive axillae (macro +micrometastases) was found by ultrasound and FNA (37/121). 41% of all axillae containing macrometastases was found by ultrasound and FNA (36/87). SLNPs were reduced by 14% (37/268). Maximum cortex thickness is the main feature to predict metastatic involvement (area under Receiver Operating Characteristic (ROC) curve (A(Z))=0.87). PMID- 12736107 TI - Acute toxicity of concurrent adjuvant radiotherapy and chemotherapy (CMF or AC) in breast cancer patients. a prospective, comparative, non-randomised study. AB - The concurrent administration of adjuvant chemotherapy and radiotherapy in breast cancer treatment might lead to an increased incidence of side-effects. In this prospective, non-randomised, comparative study, the acute toxicity of radiotherapy alone (RT) and radiotherapy concurrent with doxorubicin cyclophosphamide (AC/RT) and radiotherapy concurrent with cyclophosphamide methotrexate-5-fluorouracil (CMF/RT) was compared. We used the common toxicity criteria (CTC) to score the level of acute toxicity before, during and 6 months after the completion of the period of irradiation. The number of hospital admissions, as well as the compliance of chemotherapy, were noted. We observed that patients treated with AC/RT and CMF/RT had significant higher incidences of (high-grade) skin-toxicity, oesophagitis, dyspnoea, malaise, anorexia, nausea and hospital admission compared with those treated with RT only. The target-volume of radiotherapy was the main predictor of (high-grade) acute skin toxicity and oesophagitis. AC/RT was associated with significant more (high-grade) skin toxicity than CMF/RT. The dose of chemotherapy was reduced to less than 85% of the planned dose in 11% of patients, 17% of patients treated with concurrent chemotherapy and radiotherapy needed admission to hospital. From the results of our study, we conclude that the concurrent administration of adjuvant chemotherapy and radiotherapy leads to an unacceptably high level of acute toxicity. PMID- 12736106 TI - Differential involvement of neurotransmitters through the time course of cisplatin-induced emesis as revealed by therapy with specific receptor antagonists. AB - Advances in antiemetic therapy for chemotherapy-induced emesis have resulted in improved protection against symptoms occurring within 24 h of chemotherapy. However, the vomiting which tends to occur beyond 24 h after chemotherapy (delayed-phase vomiting) is still relatively poorly controlled by the currently available drugs, suggesting that more than one mechanism may mediate these symptoms. The standard antiemetic regimen currently recommended for prevention of chemotherapy-induced emesis includes a serotonin (5-HT(3)) antagonist and a corticosteroid. The neurokinin-1 (NK(1)) antagonist aprepitant represents a new class of antiemetic currently in clinical development. Using data obtained in 2 Phase II clinical trials of aprepitant in patients receiving chemotherapy based on the highly emetogenic chemotherapeutic agent cisplatin, we compared the time course of antiemetic effect of aprepitant, a 5-HT(3) antagonist, or a combination of both. Over the entire observation period (up to 7 days post-cisplatin), patients who received the NK(1) antagonist had a superior prevention of emesis. However, in the first 24 h after cisplatin, emesis occurred in fewer patients who received the 5-HT(3) antagonist than in patients who did not receive this class of drug. Furthermore, the majority of treatment failures in patients who received the NK(1) antagonist occurred within the first 8-12 h of chemotherapy, whereas the treatment failures in patients who received a 5-HT(3) antagonist were more evenly distributed over time. Patients who received both drugs had superior control of symptoms compared with patients who received one or the other. The difference in the time course of emesis blockade observed with two different classes of receptor antagonists provides substantial evidence for involvement of separate pathophysiological mechanisms in chemotherapy-induced vomiting. Serotonin mediates the early vomiting process that occurs within 8-12 h following cisplatin-based chemotherapy, after which time substance P acting at NK(1) receptors becomes the dominant mediator of vomiting PMID- 12736109 TI - Phase I clinical and pharmacokinetic study of E7070, a novel sulfonamide given as a 5-day continuous infusion repeated every 3 weeks in patients with solid tumours. A study by the EORTC Early Clinical Study Group (ECSG). AB - A single-agent dose-escalating phase I study on the novel sulfonamide E7070 was performed to determine the toxicity profile and the recommended dose for phase II studies. The pharmacokinetic profile of E7070 was also determined. E7070 was administered as a continuous infusion over 5 days repeated every 3 weeks. 27 patients were treated at doses ranging from 6 to 200 mg/m(2)/day. As with other administration schedules, the dose-limiting toxicities were dose-dependent, reversible neutropenia and thrombocytopenia. Although no objective responses were observed, seven patients had stable disease. E7070 displayed a non-linear pharmacokinetic profile, especially at dose-levels greater than 24 mg/m(2)/day, with a reduction in clearance and an increase in the half-life at the higher dose levels. The risk of myelosuppression became significant with an AUC greater than 4000 microg h/ml. The recommended dose of E7070 for further studies is 96 mg/m(2)/day when administered on a 5-day continuous infusion schedule every 3 weeks. PMID- 12736108 TI - Prognostic factors for survival after neoadjuvant chemotherapy in operable breast cancer. the role of clinical response. AB - The aim of this retrospective study was to assess predictive factors for clinical response to preoperative chemotherapy and prognostic factors for survival. From 1981 to 1992, 936 patients with T2-T3, N0-N1 breast cancer who received 2-6 months (median 4) of preoperative chemotherapy were selected from the Institute Curie database. Preoperative treatment was followed by surgery and/or radiotherapy. Median follow-up was 8.5 years (range 7-211 months). The objective response rate before surgery and/or radiotherapy was 58.3%. In stepwise multivariate analysis (Cox model), favourable prognostic factors for survival were the absence of pathological axillary lymph node involvement (Relative Risk (RR) 1.54; P=0.0004), low histological tumour grade (RR=1.54; P=0.0017), clinical response to preoperative chemotherapy (RR=1.45, P=0.0013), positive progesterone receptor (PR) status (RR=1.56; P=0.0001), smaller tumour size (RR=1.37; P=0.005) and lack of clinical lymph node involvement (RR=1.42; P=0.007). The association of clinical tumour response with survival is independent of the baseline characteristics of the tumour. Clinical response could be used as a surrogate marker for evaluation of the efficacy of neoadjuvant chemotherapy before assessment of the pathological response. PMID- 12736110 TI - Population pharmacokinetic modelling of unbound and total plasma concentrations of paclitaxel in cancer patients. AB - The aim of this study was to validate and further develop a mechanism-based population pharmacokinetic model for paclitaxel (Taxol; Bristol-Myers Squibb Co, Princeton, NJ, USA) based on the knowledge of Cremophor EL (CrEL) micelle entrapment and to evaluate the exposure/toxicity relationships. Paclitaxel (total and unbound) and CrEL concentrations were obtained according to a sparse sampling scheme with on average only 3.5 samples per course from 45 patients with solid tumours who received 3-hour infusions of paclitaxel (final dose range 112-233 mg/m(2)). The present data were predicted well by the mechanism-based model. In addition, bilirubin and body size were found to be significant as covariates. A change in body surface area (BSA) of 0.1 m(2) typically caused a change in clearance (CL) of 22.3 l/h and an increase in bilirubin of 10 microM typically caused a decrease in CL of 41 l/h. Toxicity was best described by a threshold model. In conclusion, even with a sparse sampling scheme, the same mechanism based binding components as in the previously developed model could be identified. Once the CrEL and total paclitaxel plasma concentrations are known, the unbound concentrations, which are more closely related to the haematological toxicity, can be predicted. PMID- 12736111 TI - Bleomycin, vincristine, lomustine and dacarbazine (BOLD) in combination with recombinant interferon alpha-2b for metastatic uveal melanoma. AB - This EORTC multicentre study analysed the efficacy and tolerability in patients with metastatic uveal melanoma of BOLD chemotherapy in combination with recombinant interferon alpha-2b. The dose of bleomycin was 15 mg on days 2 and 5, of vincristine 1 mg/m(2) on days 1 and 4, of lomustine 80 mg on day 1, and of dacarbazine (DTIC) 200 mg/m(2) on days 1-5, given every 4 weeks for a minimum of two cycles. Subcutaneous (s.c.) interferon alpha-2b at a dose of 3 x 10(6) IU was initiated on day 8 of the first cycle, and continued at a dose of 6 x 10(6) IU three times per week after 6 weeks. A median of two cycles were administered to 24 patients (median age 60.5 years). None achieved an objective response (0%; 95% Confidence Interval (CI): 0-14), 2 (8.3%) remained stable, 20 showed progression, and 2 (8.3%) were invaluable. The median progression-free survival was 1.9 months (95% CI: 1.8-3.4) and overall survival 10.6 months (95% CI: 6.9-16.4). Overall survival improved with increasingly favourable pretreatment characteristics (median, 14.7 versus 6.9 versus 6.0 months for Helsinki University Central Hospital (HUCH) Working Formulation stages IVBa, IVBb and IVBc, respectively; P=0.018). Grade 3 alopecia and neurotoxicity occurred in 13% of the patients. This multicentre study did not confirm earlier reports that BOLD with human leucocyte or recombinant interferon would induce at least 15% objective responses in metastatic uveal melanoma. PMID- 12736112 TI - A randomised study of protracted venous infusion of 5-fluorouracil (5-FU) with or without bolus mitomycin C (MMC) in patients with carcinoma of unknown primary. AB - No standard regimen has been identified for patients with a carcinoma of unknown primary (CUP). This study compared protracted venous infusion 5-fluorouracil (PVI 5-FU) with or without mitomycin C (MMC) in patients with CUP in a multicentre, prospectively randomised study. 88 patients were randomised to PVI 5-FU (300 mg/m(2)/day for a maximum of 24 weeks) +/-MMC (7 mg/m(2) 6 weekly for four courses). The overall response rate was 11.6% for PVI 5-FU alone compared with 20.0% for PVI 5-FU plus MMC (P=0.29). Median failure-free survival (FFS) was 4.1 months for PVI 5-FU and 3.6 months for PVI 5-FU plus MMC (P=0.78) with an equivalent overall survival (OS) (6.6 versus 4.7 months, P=0.60). Symptomatic benefit was observed in most patients in each arm. PVI 5-FU is a well tolerated outpatient treatment regimen for patients with CUP, although the addition of MMC provides little extra benefit. PVI 5-FU may be a potential reference regimen in randomised trials with newer chemotherapy agents in patients with CUP. PMID- 12736113 TI - Lack of impact of platinum dose intensity on the outcome of ovarian cancer patients. 10-year results of a prospective randomised phase III study comparing carboplatin-cisplatin with cyclophosphamide-cisplatin. AB - This prospective multicentre phase III trial was conducted to assess whether increased platinum dose intensity (DI) by combining carboplatin with cisplatin has an impact on overall survival (OS) and progression-free interval (PFI) compared with the standard combination of cyclophosphamide and cisplatin in patients with epithelial ovarian cancer. A total of 253 patients with epithelial ovarian cancer of stages International Federation of Gynecology and Obstetrics (FIGO) IC-IV were randomised to receive either cyclophosphamide (600 mg/m(2), intravenously (i.v.), day 1) and cisplatin (100 mg/m(2), i.v., day 2) (n=125) as the standard regimen or carboplatin (300 mg/m(2), i.v., day 1) and cisplatin (100 mg/m(2), i.v., day 2) (n=128), every 28 days for six courses. The median follow up was 6.0 years. 124 patients randomised to the platinum dose-intensified arm and 123 patients randomised to the standard arm met all of the eligibility criteria. Patient characteristics were well balanced between the two treatment groups. All eligible patients randomised were included in the analysis of OS and PFI. The median OS of the standard and platinum dose-intensified arms were 41.2 (95% Confidence Interval (CI): 29.2-50.7) and 43.0 months (95% CI: 34.3-63.2), respectively (P=Non-significant (N.S.). The median PFI in the standard arm was 29.7 (95% CI: 17.4-41.7) versus 23.1 months (95% CI: 17.8-35.4) in the platinum dose-intensified arm, respectively (P=N.S.). Toxicity, comprising leucopenia, granulocytopenia, thrombocytopenia, anaemia, emesis and nausea, was statistically significantly higher in the platinum dose-intensified arm than in the standard arm. Unexpectedly, no statistically significant differences were found between the 2 arms' overall neuro- and ototoxicity. When converting carboplatin-platinum into cisplatin-platinum on the basis of an equivalence ratio of 4:1, patients in the platinum dose-intensified arm received a total platinum dose 1.58 times the platinum dose of the standard arm. With 35.0 mg/m(2)/week being administered, the total platinum DI of the dose-intensified arm was statistically significantly (P<0.0001) higher than that of the standard regimen (with 22.0 mg/m(2) being administered). Calculating the average administered relative dose intensities of the regimens yielded almost identical results with 0.56 and 0.58 for the standard and experimental arms, respectively. Thus, by conventional means, a 1.6-fold increase in the platinum DI could be reached by combining carboplatin and cisplatin without unacceptable morbidity. Nevertheless, this did not translate into any therapeutic benefit for the patient, even in the optimally debulked group of patients for whom dose-intensification would have been expected to be of benefit. PMID- 12736114 TI - Phase II study of fotemustine in patients with advanced ovarian carcinoma. A trial of the EORTC Gynecological Cancer Group. AB - 30 patients with advanced ovarian cancer, all platinum pretreated, were treated with an induction cycle of fotemustine. Maintenance therapy was given to 6 patients. No objective response was observed among the 21 evaluable patients. The main toxicities were gastrointestinal, with grade 3 nausea and vomiting reported in 40% of the patients, and haematological, with grade 4 leucopenia reported in 2 patients and grade 4 thrombocytopenia in 5 patients. Therefore, no role has been demonstrated in our cohort for the use of fotemustine, a nitrosourea, in pretreated ovarian cancer. PMID- 12736115 TI - Loss of hMLH1 expression correlates with improved survival in stage III-IV ovarian cancer patients. AB - Pre-clinical data suggest a relationship between DNA MisMatch Repair (MMR) system failure, particularly the inactivation of genes hMLH1 and hMSH2, and resistance to drugs like cisplatin and carboplatin. We studied the correlation between loss of hMLH1 expression in tumour cells and clinical outcome in 38 patients with ovarian cancer, who underwent cisplatin-based chemotherapy. 19 patients (56%) showed loss of hMLH1 expression (Group A) while 15 patients (44%) showed normal hMLH1 expression (Group B). 4 patients were not evaluable for hMLH1 expression. The 2 groups of patients were similar for clinical characteristics, response to chemotherapy and time to progression. Group A patients showed a median survival of 55 months whereas Group B patients had a median survival of 12 months (P=0.014). Loss of hMLH1 expression was the only independent predictor of survival in the multivariate analysis. Our observations suggest a relationship between loss of hMLH1 and improved survival in advanced ovarian cancer. PMID- 12736116 TI - Organisation of follow-up in paediatric oncology. PMID- 12736118 TI - Cigarette smoking and the risk of invasive epithelial ovarian cancer in a prospective cohort study. AB - Few cohort studies have examined the association between cigarette smoking and ovarian cancer risk, either overall, or by histological subtype. In relation to the latter, it has been suggested that mucinous ovarian tumours may be aetiologically unrelated to the other types of epithelial tumours and that their respective associations with cigarette smoking may differ. We examined the association between smoking and ovarian cancer risk using data from participants in a randomised controlled trial of screening for breast cancer involving 89,835 women aged 40-59 years at recruitment. Cox proportional hazards models were used to estimate rate ratios (RR) and 95% confidence intervals (CI). During an average of 16.5 years of follow-up, we observed 454 incident cases of ovarian cancer (184 serous, 67 endometrioid, 32 mucinous, 171 other or unknown). We found that women who had smoked for several decades had an approximately two-fold increased risk of epithelial ovarian cancer. Relative to never-smokers, women who had smoked for 40 years or more were at the highest risk (RR=2.50, 95% CI=1.37-4.56). The association with non-mucinous tumours was similar to that observed overall. For mucinous tumours, a two-fold increased risk was observed with smoking of shorter duration, although the number of mucinous tumours in our data-set was small. Long term cigarette smoking may be associated with an increased risk of epithelial ovarian tumours. PMID- 12736119 TI - Histone acetylation-mediated regulation of genes in leukaemic cells. AB - Histone deacetylase (HDAC) and histone acetyltransferase (HAT) functions are associated with various cancers, and the inhibition of HDAC has been found to arrest disease progression. Here, we have investigated the gene expression profiles of leukaemic cells in response to the HDAC inhibitor trichostatin A (TSA) using oligonucleotide microarrays. Nucleosomal histone acetylation was monitored in parallel and the expression profiles of selected genes were confirmed by quantitative polymerase chain reaction (PCR). A large number of genes (9% of the genome) were found to be similarly regulated in CCRF-CEM and HL 60 cells in response to TSA, and genes showing primary and secondary responses could be distinguished by temporal analysis of gene expression. A small fraction of genes were highly sensitive to histone hyper-acetylation, including XRCC1, HOXB6, CDK10, MYC, MYB, NMI and CBFA2T3 and many were trans-acting factors relevant to cancer. The most rapidly repressed gene was MKRN3, an imprinted gene involved in the Prader-Willi syndrome. PMID- 12736121 TI - Control of Rapid Arm Movements When Target Position Is Altered During Saccadic Suppression. AB - This experiment examined whether rapid arm movements can be corrected in response to a change in target position that occurs just prior to movement onset, during saccadic suppression of displacement. Because the threshold of retinal input reaches its highest magnitude at that time, displacement of the visual target of a saccade is not perceived. Subjects (N = 6) were instructed to perform very rapid arm movements toward visual targets located 16, 20, and 24 degrees from midline (on average, movement time was 208 ms). On some trials the 20 degrees target was displaced 4 degrees either to the right or to the left during saccadic suppression. For double-step trials, arm movements did not deviate from their original trajectory. Movement endpoints and movement structure (i.e., velocity and acceleration-time profiles) were similar whether or not target displacements occurred, showing the failure of proprioceptive signals or internal feedback loops to correct the arm trajectory. Following this movement, terminal spatially oriented movements corrected the direction of the initial movement (as compared with the single-step control trials) when the target eccentricity decreased by 4 degrees. Subjects were unaware of these spatial corrections. Therefore, spatial corrections of hand position were driven by the goal level of the task, which was updated by oculomotor corrective responses when a target shift occurred. PMID- 12736122 TI - Asymmetries in Coupling Dynamics of Perception and Action. AB - The influence of information-based dynamics on coordination dynamics of rhythmic movement was examined with special reference to the expression of asymmetries. In Experiment 1, right-handed subjects performed unimanual, rhythmical movements in coordination with either a discrete or continuous visual display. The right hand visual display system defined a more stable perception-action collective than the left, particularly when continuous visual information was available. In Experiment 2, the same subjects performed rhythmic bimanual movements in coordination with a continuous visual display. The action collective was inherently more stable than the perception-action collective, although similar patterns were observed at both levels. Importantly, the dynamics of the perception-action collective impinged upon the dynamics of the action collective in terms of stability. Asymmetries remained evident between limbs in the bimanual preparations, with the left hand exhibiting greater limit-cycle variability and also a tendency to more often effect transitions at the action couple. Features of dynamical models that capture characteristics of manual asymmetries are discussed. PMID- 12736120 TI - NF-kappa B activation in vivo in both host and tumour cells by the antivascular agent 5,6-dimethylxanthenone-4-acetic acid (DMXAA). AB - 5,6-Dimethylxanthenone-4-acetic acid (DMXAA), a new anticancer agent developed in this centre, has an antivascular action and causes regression of transplantable murine tumours that is mediated partially by the intratumoral production of tumour necrosis factor (TNF). DMXAA activates the nuclear factor-kappaB (NF kappaB) transcription factor, which is involved in TNF synthesis and has also been suggested to mediate resistance to TNF. We wished to determine whether tumour cell NF-kappaB activation modulated the in vitro and in vivo effects of DMXAA. We compared the response of the 70Z/3 pre-B lymphoma cell line with that of its mutant 1.3E2 sub-line, which has a defective gamma-subunit of IKK, the kinase that phosphorylates IkappaB leading to NF-kappaB activation. As shown by electrophoretic mobility shift assays (EMSAs), DMXAA induced in vitro translocation of NF-kappaB (p50 and p65 subunits) into the nucleus of 70Z/3 cells, but not of 1.3E2 cells. However, when the cell lines were then grown as subcutaneous tumours in mice and treated with DMXAA (25 mg/kg), activation of NF kappaB was found in nuclear extracts prepared from both 70/Z3 and 1.3E2 tumours, as well as from Colon 38 tumours that were used for comparison. This suggests that DMXAA induces NF-kappaB responses in host components of the tumour. Tumours grown from both 70Z/3 and 1.3E2 cells were found to regress completely following DMXAA treatment. Thus, the antitumour action of DMXAA appears to be independent of the ability of the target tumour cell population to induce NF-kappaB expression. Moreover, activation of NF-kappaB in the tumour cell did not confer resistance to DMXAA-induced therapy. PMID- 12736123 TI - An Energetic Comparison of Symmetrical and Asymmetrical Human Gait. AB - This article contrasts the mechanical energy profiles of asymmetrical galloping with those of symmetrical running in adult humans. Seven female subjects were filmed while performing overground running and galloping at their preferred velocities. A previous study (Whitall & Caldwell, 1992) showed that kinematic differences between these gait modes included higher preferred velocity for running than galloping, with distinct differences in interlimb coordination but surprisingly similar intralimb patterns. Energetically, in the present study the whole body center of mass during galloping was found to behave much as it does in walking; kinetic and potential energy profiles were out of phase, as compared with running, which exhibited in-phase fluctuations of kinetic and potential energies. The primary reason for these center of mass differences was found in the energetics of the back leg of galloping, which demonstrated alterations in timing of its energy fluctuations and less energy generation than the front leg. Analysis of the power sources underlying the segmental energies during swing phase showed that the back leg's energy changes were accomplished mainly through reduced use of the hip muscles and less interlimb energy transfer. The back leg's energetics during swing also displayed a shift toward greater reliance on nonmuscular energy sources. A pattern of energy inflow during early swing and energy outflow during late swing was common to both running and galloping, although the galloping legs both demonstrated more abrupt transitions between these phases. The possibility is raised that the 67/33 interlimb phasing ratio used in galloping is selected to reduce mechanical energy variations of the total body center of mass. These data suggest that models of asymmetric gait in humans must account for more than merely phase alteration. PMID- 12736124 TI - Human Eye Movements During Visually Guided Stepping. AB - Visually guided locomotion was studied in an experiment in which human subjects (N = 8) had to accurately negotiate a series of irregularly spaced stepping stones while infrared reflectometry and electrooculography were used to continuously record their eye movements. On average, 68% of saccades made toward the next target of footfall had been completed (visual target capture had occurred) while the foot to be positioned was still on the ground; the remainder were completed in the first 300 ms of the swing phase. The subjects' gaze remained fixed on a target, on average, until 51 ms after making contact with it, with little variation. A greater amount of variation was seen in the timing of trailing footlift relative to visual target capture. Assuming that subjects sampled the visual cues as and when they were required, visual information appeared most useful when the foot to be positioned was still on the ground. PMID- 12736125 TI - Energetic Cost and Stability During Human Walking at the Preferred Stride Velocity. AB - The possibility that preferred modes of locomotion emerge from dynamical and optimality constraints and the energetic and dynamical constraints on preferred and predicted walking frequency are explored in this article. Participants were required to walk on a treadmill at their preferred frequency, at a frequency predicted as the resonance of a hybrid pendulum-spring model of the legs, and at frequencies +/-15%, +/-25%, +/-35% of the predicted frequency. Walking at the preferred and predicted frequencies resulted in minimal metabolic costs and maximal stability of the head and joint actions. Mechanical energy conservation was constant across conditions. The head was more stable than the joints. The joints appeared to be in service of the head in maintaining a stable trajectory. The major findings of this study suggest a complementary relationship between energetic (physiological) and stability constraints in the adoption of a preferred frequency of walking. Multiple subsystems may be involved in constraining observed macroscopic behavior in intact biological systems. The approach and results of the study imply that a useful tack in understanding how dynamical control structures arise is to study the potential criteria that serve to act as constraints on skilled movement patterns in unimpaired and impaired populations. PMID- 12736126 TI - The Organization of Action Sequences: Evidence From a Relearning Task. AB - In two studies, the organization of sequential behavior in transcription typing was investigated. The design of the studies made it possible to test the hypothesis that sequential skill in typing resides only at an abstract, effector independent level. Skilled typists (N = 12) learned to type on an altered keyboard in an experimental paradigm that allowed only certain components of the motor control system to adapt to the alterations. When performance was compared on a pretest and a posttest, various decrements in the typists' speed and accuracy were observed. The forms of these decrements provided evidence against a strong form of the effector-independent hypothesis. PMID- 12736127 TI - The Topology of Limb Deceleration in Prehension Tasks. AB - Whether a subject uses a constant tau-dot strategy to decelerate a reach to grasp an object is typically determined by performing a linear regression of tau onto real time to contact. This method has the disadvantage of requiring an assumption (on the part of the experimenter) concerning the endpoint targeted by the subject. In the present contribution, an alternative, topological method is proposed that makes use of a phase-plane representation of the movement. In two experiments, experimentally obtained movement trajectories were evaluated, using this new method, and the results were found to support the constant tau-dot strategy. At the same time, however, an alternative model, associated with dynamical systems theory, was demonstrated to fit the data even better. It is argued that the topology of the phase plane suggests a softening spring description for a large part of the decelerative phase, followed by a final adjustment. Deterioration of the functionality of the adjustment made in this latter phase was observed when visual information was not available during movement execution. PMID- 12736128 TI - Effects of Pointing Rate and Availability of Visual Feedback on Visual and Proprioceptive Components of Prism Adaptation. AB - While looking through laterally displacing prisms, subjects pointed 60 times straight ahead of their nose at a rate of one complete movement every 2 or 3 s, with visual feedback available early in the pointing movement or delayed until the end of the movement. Sagittal pointing was paced such that movement speed covaried with pointing rate. Aftereffect measures (obtained after every 10 pointing trials) showed that when the limb became visible early in a pointing movement, proprioceptive adaptation was greater than visual, but when visual feedback was delayed until the end of the movement, the reverse was true. This effect occurred only with the 3-s pointing rate, however. With the 2-s pointing rate, adaptation was predominately proprioceptive in nature, regardless of feedback availability. Independent of the availability of visual feedback, visual adaptation developed more quickly with 3-s pointing, whereas proprioceptive adaptation developed more rapidly with 2-s pointing. These results are discussed in terms of a model of perceptual-motor organization in which the direction of coordinative (guidance) linkage between eye-head (visual) and hand-head (proprioceptive) systems (and consequently the locus of discordance registration and adaptive recalibration) is determined jointly by pointing rate and feedback availability. An additional effect of pointing rate is to determine the rate of discordant inputs. Maximal adaptive recalibration occurs when the input (pointing) rate matches the time constant of the adaptive encoder in the guided system. PMID- 12736129 TI - Cognitive Load and Prism Adaptation. AB - Subjects wore goggles with prisms that laterally displaced the visual field (rightward by 11.4 degree) and with full view of the limb engaged in paced (2-s rate) sagittal pointing at either an implicit ("straight ahead of the nose") target (Experiment 1) or an explicit (positioned leftward by 11.4 degree) target (in Experiment 2). In experimental conditions, subjects performed a secondary cognitive task (mental arithmetic) simultaneously during target pointing. In control conditions, no cognitive load was imposed. Aftereffect measures of adaptation to the prismatic displacement were not substantially different when problem solving was required, but terminal error of the exposure pointing task was reliably affected by cognitive load. These results are consistent with the hypothesis of separable mechanisms for adaptive coordination and adaptive alignment. Adaptive coordination may be mediated by strategically flexible coordinative linkage between sensory motor systems (eye-head and hand-head), but spatial alignment seems to be mediated by adaptive encoders within coordinatively linked subsystems. If the coordination task involves predominately automatic processing, coordinative linkage can be frequent enough under cognitive load for substantial realignment to occur even though exposure performance (adaptive coordination) may be less than optimal. PMID- 12736130 TI - Pew (1966) Revisited: Acquisition of Hierarchical Control as a Function of Observational Practice. AB - Two experiments are reported that utilized a task similar to one developed by Pew (1966) in which subjects controlled the position of a continually moving cursor by alternately pressing two response keys. Experiments I and 2 replicated Pew's findings. That is, over acquisition trials, subjects exhibited an increase in their rate of responding, an increase in response rhythmicity, and a reduction in error. After a 24-hr retention interval, it was demonstrated that the rate and rhythmicity of responding for the observation practice groups were indistinguishable from those of the actual practice groups. In general, however, as the demands of the task (cursor acceleration rate and/or target amplitude) were increased, there was a tendency for the absolute differences in error between the actual and observation practice groups to increase, These data support the notion that subjects may choose appropriate response/control schemes (i.e., open-loop and hierarchical control) without actual practice experiences. PMID- 12736131 TI - Augmented Kinematic Feedback for Motor Learning. AB - Although the study of feedback about goal achievement (knowledge of results, KR) has been important for the development principles of augmented information feedback in simple skills, there is reason to question the generalizability of these findings to many common learning situations. A more appropriate type of information for skill learning appears to be augmented kinematic (or kinetic) feedback regarding the movement pattern. The experiments presented here extend recent findings about KR to a paradigm involving kinematic feedback. In Experiment 1, we examined how several kinds of temporal and spatial kinematic information supplement KR in learning. Spatial kinematic variables were more effective than temporal variables, as indicated by performance in a retention test without kinematic feedback. In Experiment 2, we manipulated the schedule of augmented kinematic feedback in a method that paralleled previous KR work. We contrasted averaged schedules of augmented feedback, in which information was given either after every trial or as averaged information after every set of five trials. On retention tests without kinematic feedback given 1 day and 1 week after acquisition, averaged schedules led to enhanced performance over an every trial format. Together, these results begin to define the variables important in kinematic feedback, and suggest that this feedback may influence learning in ways parallel to KR. PMID- 12736132 TI - Interference Between Location and Distance Information in Motor Short-Term Memory: The Respective Roles of Direct Kinesthetic Signals and Abstract Codes. AB - Interference between location and distance information in motor short-term memory has been hypothesized on the basis of the systematic pattern of undershooting and overshooting in movement reproduction that occurs when the starting position for reproduction movements is shifted. To determine the possible contribution of limb specific kinesthetic information to this systematic undershooting-overshooting pattern, we compared the reproduction of linear arm positioning movements performed under either same-limb or switched-limb conditions. Ten subjects were assigned to either a location or distance cue condition, and each subject completed a total of 40 trials, 20 under same-limb and 20 under switched-limb conditions. Each trial consisted of criterion and reproduction movements, separated by a 10-s retention interval. The starting position for the reproduction movement was shifted by 0, 2, or 4 cm in either direction from that of the criterion movement. The systematic undershooting-overshooting pattern, which occurs when either the movement location or distance is reproduced, arose under both the same-limb and switched-limb conditions, suggesting that the primary cause of the location-distance interference is not limb-specific kinesthetic information. Rather, more abstract information in the form of a conceptual memory code appears to be the probable cause of the location distance interference phenomenon. PMID- 12736133 TI - Evidence of Common Timing Processes in the Control of Manual, Orofacial, and Speech Movements. AB - Recent investigations of timing in motor control have been interpreted as support for the concept of brain modularity. According to this concept, the brain is organized into functional modules that contain mechanisms responsible for general processes. Keele and colleagues (Keele & Hawkins, 1982; Keele & Ivry, 1987; Keele, Ivry, & Pokorny, 1987; Keele, Pokorny, Corcos, & Ivry, 1985) demonstrated that the within-subject variability in. cycle duration of repetitive movements is correlated across finger, forearm, and foot movements, providing evidence in support of a general timing module. The present study examines the notion of timing modularity of speech and nonspeech movements of the oral motor system as well as the manual motor system. Subjects produced repetitive movements with the finger, forearm, and jaw. In addition, a fourth task involved the repetition of a syllable. All tasks were to be produced with a 400-ms cycle duration; target duration was established with a pacing tone, which then was removed. For each task, the within-subject variability of the cycle duration was computed for the unpaced movements over 20 trials. Significant correlations were found between each pair of effectors and tasks. The present results provide evidence that common timing processes are involved not only in movements of the limbs, but also in speech and nonspeech movements of oral structures. PMID- 12736134 TI - Recovery of Locomotion in Monkeys With Spinal Cord Lesions. AB - This study reanalyzes kinematically (via film) the pre- and postoperative locomotor behavior of 4 of the 10 monkeys with partial spinal cord lesions (T8) briefly described by Eidelberg, Walden, and Nguyen (1981). The behavior of the remaining 6 monkeys is qualitatively described. The analysis reveals that 5 of the animals initially exhibited unilateral hind limb stepping. Hind and forelimb cycle durations often differed postoperatively; the hind limbs commonly showed increased values, whereas forelimb cycle durations were reduced: ipsilateral interlimb phase values were usually inconsistent. A review of prior studies of primate spinal cord lesions indicates that sparing of the ventrolateral quadrant may not be essential for locomotor recovery (cf. Eidelberg, Walden, & Nguyen, 1981). Furthermore, this review as well as the kinematic analysis indicates that primates with very significant spinal lesions can stilI exhibit locomotor movements. Thus, although the primate's spinal cord seems less able than other mammals' to readily organize locomotor movements (Eidelberg, Walden, & Nguyen, 1981), the total absence of stepping in primates with completely transected cords is unexpected and warrants further research. PMID- 12736135 TI - An ecdysone and tetracycline dual regulatory expression system for studies on Rac1 small GTPase-mediated signaling. AB - Regulated expression systems are invaluable for studying gene function, offer advantages of dosage-dependent and temporally defined gene expression, and limit possible clonal variation when toxic or pleiotropic genes are overexpressed. Previously, establishment of inducible expression systems, such as tetracycline- and ecdysone-inducible systems, required assessment of the inducible characteristics of individual clones by tedious luciferase assays. Taking advantage of a green fluorescent protein (GFP) reporter controlled by tetracycline- or ecdysone-responsive element and fluorescence-activated cell sorting, we propose a simple and efficient strategy to select highly inducible cell lines according to their fluorescence profiles after transiently transfecting the candidate cell pools with a surrogate GFP reporter. We have demonstrated that tetracycline- and ecdysone-inducible systems could be set up in Madin-Darby canine kidney and HEK-293 cells by employing this selection scheme. Importantly, this dual regulatory expression system is applied in studying the complex interplay between two Ras-related small GTPases, Cdc42 and Rac1, on detachment-induced apoptosis. Furthermore, establishment of two tightly regulated expression systems in one target cell line could be of great advantage for dissecting small GTPase Rac1-transduced signaling pathways by using global gene expression approaches such as proteomic assays. PMID- 12736136 TI - Identification of an apical Cl-/HCO-3 exchanger in rat kidney proximal tubule. AB - SLC26A6 (or putative anion transporter 1, PAT1) is located on the apical membrane of mouse kidney proximal tubule and mediates Cl-/HCO3- exchange in in vitro expression systems. We hypothesized that PAT1 along with a Cl-/HCO3- exchange is present in apical membranes of rat kidney proximal tubules. Northern hybridizations indicated the exclusive expression of SLC26A6 (PAT1 or CFEX) in rat kidney cortex, and immunocytochemical staining localized SLC26A6 on the apical membrane of proximal tubules, with complete prevention of the labeling with the preadsorbed serum. To examine the functional presence of apical Cl-/HCO3 exchanger, proximal tubules were isolated, microperfused, loaded with the pH sensitive dye BCPCF-AM, and examined by digital ratiometric imaging. The pH of the perfusate and bath was kept at 7.4. Buffering capacity was measured, and transport rates were calculated as equivalent base flux. The results showed that in the presence of basolateral DIDS (to inhibit Na+-HCO3- cotransporter 1) and apical EIPA (to inhibit Na+/H+ exchanger 3), the magnitude of cell acidification in response to addition of luminal Cl- was approximately 5.0-fold higher in the presence than in the absence of CO2/HCO3-. The Cl--dependent base transport was inhibited by approximately 61% in the presence of 0.5 mM luminal DIDS. The presence of physiological concentrations of oxalate in the lumen (200 microM) did not affect the Cl-/HCO3- exchange activity. These results are consistent with the presence of SLC26A6 (PAT1) and Cl-/HCO3- exchanger activity in the apical membrane of rat kidney proximal tubule. We propose that SLC26A6 is likely responsible for the apical Cl-/HCO3- (and Cl-/OH-) exchanger activities in kidney proximal tubule. PMID- 12736137 TI - G alpha 13-mediated transformation and apoptosis are permissively dependent on basal ERK activity. AB - We previously reported that the alpha-subunit of heterotrimeric G13 protein induces either mitogenesis and neoplastic transformation or apoptosis in a cell dependent manner. Here, we analyzed which signaling pathways are required for G alpha 13-induced mitogenesis or apoptosis using a novel mutant of G alpha 13. We have identified that in human cell line LoVo, the mutation encoding substitution of Arg260 to stop codon in mRNA of G alpha 13 subunit produced a mutant protein (G alpha 13-T) that lacks a COOH terminus and is endogenously expressed in LoVo cells as a polypeptide of 30 kDa. We found that G alpha 13-T lost its ability to promote proliferation and transformation but retained its ability to induce apoptosis. We found that full-length G alpha 13 could stimulate Elk1 transcription factor, whereas truncated G alpha 13 lost this ability. G alpha 13 dependent stimulation of Elk1 was inhibited by dominant-negative extracellular signal-regulated kinase (MEK) but not by dominant-negative MEKK1. Similarly, MEK inhibitor PD-98059 blocked G alpha 13-induced Elk1 stimulation, whereas JNK inhibitor SB-203580 was ineffective. In Rat-1 fibroblasts, G alpha 13-induced cell proliferation and foci formation were also inhibited by dominant-negative MEK and PD-98059 but not by dominant-negative MEKK1 and SB-203580. Whereas G alpha 13-T alone did not induce transformation, coexpression with constitutively active MEK partially restored its ability to transform Rat-1 cells. Importantly, full-length but not G alpha 13-T could stimulate Src kinase activity. Moreover, G alpha 13-dependent stimulation of Elk1, cell proliferation, and foci formation were inhibited by tyrosine kinase inhibitor, genistein, or by dominant-negative Src kinase, suggesting the involvement of a Src-dependent pathway in the G alpha 13-mediated cell proliferation and transformation. Importantly, truncated G alpha 13 retained its ability to stimulate apoptosis signal-regulated kinase ASK1 and c Jun terminal kinase, JNK. Interestingly, the apoptosis induced by G alpha 13-T was inhibited by dominant-negative ASK1 or by SB-203580. PMID- 12736138 TI - Glutamate 59 is critical for transport function of the amino acid cotransporter KAAT1. AB - KAAT1 is a neutral amino acid transporter activated by K+ or by Na+ (9). The protein shows significant homology with members of the Na+/Cl--dependent neurotransmitter transporter super family. E59G KAAT1, expressed in Xenopus oocytes, exhibited a reduced leucine uptake [20-30% of wild-type (WT)], and kinetic analysis indicated that the loss of activity was due to reduction of Vmax and apparent affinity for substrates. Electrophysiological analysis revealed that E59G KAAT1 has presteady-state and uncoupled currents larger than WT but no leucine-induced currents. Site-directed mutagenesis analysis showed the requirement of a negative charge in position 59 of KAAT1. The analysis of permeant and impermeant methanethiosulfonate reagent effects confirmed the intracellular localization of glutamate 59. Because the 2-aminoethyl methanethiosulfonate hydrobromid inhibition was not prevented by the presence of Na+ or leucine, we concluded that E59 is not directly involved in the binding of substrates. N-ethylmaleimide inhibition was qualitatively and quantitatively different in the two transporters, WT and E59G KAAT1, having the same cysteine residues. This indicates an altered accessibility of native cysteine residues due to a modified spatial organization of E59G KAAT1. The arginine modifier phenylglyoxal effect supports this hypothesis: not only cysteine but also arginine residues become more accessible to the modifying reagents in the mutant E59G. In conclusion, the results presented indicate that glutamate 59 plays a critical role in the three-dimensional organization of KAAT1. PMID- 12736139 TI - Hyperoxia induces retinal vascular endothelial cell apoptosis through formation of peroxynitrite. AB - Hyperoxia exposure induces capillary endothelial cell apoptosis in the developing retina, leading to vaso-obliteration followed by proliferative retinopathy. Previous in vivo studies have shown that endothelial nitric oxide synthase (NOS3) and peroxynitrite are important mediators of the vaso-obliteration. Now we have investigated the relationship between hyperoxia, NOS3, peroxynitrite, and endothelial cell apoptosis by in vitro experiments using bovine retinal endothelial cells (BREC). We found that BREC exposed to 40% oxygen (hyperoxia) for 48 h underwent apoptosis associated with activation of caspase-3 and cleavage of the caspase substrate poly(ADP-ribose) polymerase. Hyperoxia-induced apoptosis was associated with increased formation of nitric oxide, peroxynitrite, and superoxide anion and was blocked by treatment with uric acid, nitro-L-arginine methyl ester, or superoxide dismutase. Analyses of the phosphatidylinositol 3 kinase/Akt kinase survival pathway in cells directly treated with peroxynitrite revealed inhibition of VEGF- and basic FGF-induced activation of Akt kinase. These results suggest that hyperoxia-induced formation of peroxynitrite induces BREC apoptosis by crippling key survival pathways and that blocking peroxynitrite formation prevents apoptosis. These data may have important clinical implications for infants at risk of retinopathy of prematurity. PMID- 12736142 TI - The future of GI and liver research: editorial perspectives. III. JNK/AP-1 regulation of hepatocyte death. AB - Activation of the JNK/activator protein-1 (AP-1)-signaling pathway is a common mediator of hepatocyte death from a variety of stimuli. Although the mechanism by which JNK or AP-1 promotes death is unknown, it results when activation of this signaling pathway is unusually prolonged. Although JNK/AP-1 mediates TNF-induced cell death at or above the level of the mitochondria, the ability of JNK/AP-1 to promote death from necrosis as well as apoptosis suggests that JNK/AP-1 may induce death by several mechanisms. Recognition of JNK/AP-1 signaling as a critical promoter of hepatocyte death raises the possibility that the therapeutic manipulation of this pathway may be effective in the treatment of human liver disease. PMID- 12736140 TI - Acridine orange induces translocation of phosphatidylserine to red blood cell surface. AB - Clustering of band-3 on red blood cell (RBC) surface has been assumed to catalyze RBC phagocytosis. In studying this subject, acridine orange (AO) has commonly been employed on the assumption that it specifically induces band-3 clustering. In the present study, we show that AO strongly induces translocation of phosphatidylserine (PS) to RBC surface. Because surface PS is well known to induce RBC intercellular interaction, these findings suggest that the use of AO as a specific inducer of band-3 clustering is questionable. It is possible that band-3 clustering and PS translocation are interdependent, and this interrelationship has yet to be explored. PMID- 12736143 TI - The future of GI and liver research: editorial perspectives. IV. Visceral afferents: an update. AB - The number of articles published in American Journal of Physiology Gastrointestinal and Liver Physiology over the last 15 years on visceral afferents has increased dramatically. This reflects our growing ability to study the characteristics and function of visceral afferents and also the recognition of their importance in the maintenance of homeostasis and also in a number of pathophysiological conditions. However, there are several key unanswered questions concerning the function of visceral afferents that await further investigation. PMID- 12736144 TI - Expression and function of KCNH2 (HERG) in the human jejunum. AB - Previous studies suggest that ether-a-go-go related gene (ERG) KCNH2 potassium channels contribute to the control of motility patterns in the gastrointestinal tract of animal models. The present study examines whether these results can be translated into a role in human gastrointestinal muscles. Messages for two different variants of the KCNH2 gene were detected: KCNH2 V1 human ERG (HERG) (28) and KCNH2 V2 (HERG(USO)) (13). The amount of V2 message was greater than V1 in both human jejunum and brain. The base-pair sequence that gives rise to domains S3-S5 of the channel was identical to that previously published for human KCNH2 V1 and V2. KCNH2 protein was detected immunohistochemically in circular and longitudinal smooth muscle and enteric neurons but not in interstitial cells of Cajal. In the presence of TTX (10(-6) M), atropine (10(-6) M). and l nitroarginine (10(-4) M) human jejunal circular muscle strips contracted phasically (9 cycles/min) and generated slow waves with superimposed spikes. Low concentrations of the KCNH2 blockers E-4031 (10(-8) M) and MK-499 (3 x 10(-8) M) increased phasic contractile amplitude and the number of spikes per slow wave. The highest concentration of E-4031 (10(-6) M) produced a 10-20 mV depolarization, eliminated slow waves, and replaced phasic contractions with a small tonic contracture. E-4031 (10(-6) M) did not affect [(14)C]ACh release from enteric neurons. We conclude that KCNH2 channels play a fundamental role in the control of motility patterns in human jejunum through their ability to modulate the electrical behavior of smooth muscle cells. PMID- 12736145 TI - Glucagon-like peptide-2-enhanced barrier function reduces pathophysiology in a model of food allergy. AB - Penetration of the gut epithelial barrier by intact luminal antigen is necessary for immunologically mediated pathophysiology in the context of food allergy. We investigated if glucagon-like peptide-2 (GLP-2) could affect immediate hypersensitivity and late-phase allergic inflammation in a murine model. Mice were sensitized to horseradish peroxidase (HRP); studies were conducted 14 days later. Mice were treated with 5 microg GLP-2 subcutaneously 4 h before antigen challenge. For immediate hypersensitivity, jejunal segments in Ussing chambers were challenged by luminal HRP antigen. GLP-2 treatment reduced the uptake of HRP and the antigen-induced secretory response after luminal challenge. GLP-2 appears to reduce macromolecular uptake independent of the CD23-mediated enhanced antigen uptake pathway. For the late phase, mice were gavaged with antigen, and 48 h later the function and histology of the jejunum were examined. GLP-2 prevented the usual prolonged permeability defect and reduced the number of inflammatory cells in the mucosa. Our studies demonstrate that a single treatment of sensitized mice with GLP diminishes both immediate and late-phase hypersensitivity reactions characteristic of food allergy by inhibiting transepithelial uptake of antigen. PMID- 12736146 TI - Processing of vegetable-borne carotenoids in the human stomach and duodenum. AB - Carotenoids are thought to diminish the incidence of certain degenerative diseases, but the mechanisms involved in their intestinal absorption are poorly understood. Our aim was to obtain basic data on the fate of carotenoids in the human stomach and duodenum. Ten healthy men were intragastrically fed three liquid test meals differing only in the vegetable added 3 wk apart and in a random order. They contained 40 g sunflower oil and mashed vegetables as the sole source of carotenoids. Tomato puree provided 10 mg lycopene as the main carotenoid, chopped spinach (10 mg lutein), and carrot puree (10 mg beta carotene). Samples of stomach and duodenal contents and blood samples were collected at regular time intervals after meal intake. all-trans and cis carotenoids were assayed in stomach and duodenal contents, in the fat and aqueous phases of those contents, and in chylomicrons. The cis-trans beta-carotene and lycopene ratios did not significantly vary in the stomach during digestion. Carotenoids were recovered in the fat phase present in the stomach during digestion. The proportion of all-trans carotenoids found in the micellar phase of the duodenum was as follows (means +/- SE): lutein (5.6 +/- 0.4%), beta-carotene (4.7 +/- 0.3%), lycopene (2.0 +/- 0.2%). The proportion of 13-cis beta-carotene in the micellar phase was significantly higher (14.8 +/- 1.6%) than that of the all-trans isomer (4.7 +/- 0.3%). There was no significant variation in chylomicron lycopene after the tomato meal, whereas there was significant increase in chylomicron beta-carotene and lutein after the carrot and the spinach meals, respectively. There is no significant cis-trans isomerization of beta carotene and lycopene in the human stomach. The stomach initiates the transfer of carotenoids from the vegetable matrix to the fat phase of the meal. Lycopene is less efficiently transferred to micelles than beta-carotene and lutein. The very small transfer of carotenoids from their vegetable matrices to micelles explains the poor bioavailability of these phytomicroconstituents. PMID- 12736148 TI - The splanchnic metabolism of flavonoids highly differed according to the nature of the compound. AB - The absorption and splanchnic metabolism of different flavonoids (namely quercetin, kaempferol, luteolin, eriodictyol, genistein, and catechin) were investigated in rats after an in situ perfusion of jejunum plus ileum (14 nmol/min). Net transfer across the brush border ranged widely according to the perfused compound (from 78% for kaempferol to 35% for catechin). This variation seems linked to the lipophilicity of a given flavonoid rather than to its three dimensional structure. Except for catechin, conjugated forms of perfused flavonoids were also detected in the intestinal lumen, but the extent of this secretion depended on the nature of the perfused compounds (52% for quercetin to 11% for genistein). For some of the perfused aglycones, biliary secretion was an important excretion route: 30% of the perfused dose for genistein but only 1% for catechin. Thus the splanchnic metabolism of flavonoid is controlled by several factors: 1) the efficiency of their transfer through the brush border, 2) the intensity of the intestinal secretion of conjugates toward the mucosal and serosal sides, respectively, and 3) the biliary secretion of conjugates. These data suggested that the splanchnic metabolism of perfused flavonoids depends on the nature of the compound considered, which in turn influences their availability for peripheral tissues. PMID- 12736147 TI - S-adenosylmethionine (AdoMet) modulates endotoxin stimulated interleukin-10 production in monocytes. AB - IL-10 is produced by a large variety of cells including monocytes, macrophages, B and T lymphocytes, as well as natural killer cells and is an important suppressor for both immunoproliferative and inflammatory responses. IL-10 exerts antifibrotic effects in the liver, and decreased monocyte synthesis of IL-10 is well documented in alcoholic cirrhosis. Intracellular deficiency of S adenosylmethionine (AdoMet) is a hallmark of toxin-induced liver injury. Although the administration of exogenous AdoMet attenuates this injury, the mechanisms of its actions are not fully established. This study was performed to investigate the effect of exogenous AdoMet on IL-10 production in LPS-stimulated RAW 264.7 cells, a murine macrophage cell line. Our results demonstrated that exogenous AdoMet administration enhanced both protein production and gene expression of IL 10 in RAW 264.7 cells. Ethionine, an inhibitor for methionine adenosyltransferases, inhibited LPS-stimulated IL-10 both at the protein and mRNA levels. Exogenous AdoMet increased the intracellular cAMP concentration as early as 3 h and continued for 24 h after AdoMet treatment; however, the inhibitors for both adenylyl cyclase and PKA did not significantly affect IL-10 production. On the basis of these results, we conclude that AdoMet administration may exert its anti-inflammatory and hepatoprotective effects, at least in part, by enhancing LPS-stimulated IL-10 production. PMID- 12736149 TI - Inhibition of sustained smooth muscle contraction by PKA and PKG preferentially mediated by phosphorylation of RhoA. AB - The role of RhoA in myosin light-chain (MLC)(20) dephosphorylation and smooth muscle relaxation by PKA and PKG was examined in freshly dispersed and cultured smooth muscle cells expressing wild-type RhoA, constitutively active Rho(V14), and phosphorylation site-deficient Rho(A188). Activators of PKA (5,6-dichloro-1 beta-ribofuranosyl benzimidazole 3',5'-cyclic monophosphothionate, Sp-isomer; cBIMPS) or PKG [8-(4-chlorophenylthio)guanosine 3',5'-cyclic monophosphate (8 pCPT-cGMP), sodium nitroprusside (SNP)] or both PKA and PKG (VIP) induced phosphorylation of constitutively active Rho(V14) and agonist (ACh)- or GTPgammaS stimulated wild-type RhoA but not Rho(A188). Phosphorylation was accompanied by translocation of membrane-bound wild-type RhoA and Rho(V14) to the cytosol and complete inhibition of ACh-stimulated Rho kinase and phospholipase D activities, RhoA/Rho kinase association, MLC(20) phosphorylation, and sustained muscle contraction. Each of these events was blocked depending on the agent used, by the PKG inhibitor KT5823 or the PKA inhibitor myristoylated PKI. Inhibitors were used at a concentration (1 microM) previously shown by direct measurement of kinase activity to selectively inhibit the corresponding kinase. In muscle cells overexpressing the active phosphorylation site-deficient mutant Rho(A188), MLC(20) phosphorylation was partly inhibited by SNP, VIP, cBIMPS, and 8-pCPT cGMP, suggesting the existence of an independent inhibitory mechanism downstream of RhoA. Results demonstrate that dephosphorylation of MLC(20) and smooth muscle relaxation are preferentially mediated by PKG- and PKA-dependent phosphorylation and inactivation of RhoA. PMID- 12736150 TI - Time-dependent platelet-vessel wall interactions induced by intestinal ischemia reperfusion. AB - Platelets roll and adhere in venules exposed to ischemia-reperfusion (I/R). This platelet-endothelial adhesion may influence leukocyte trafficking because platelet depletion decreases I/R-induced leukocyte emigration. The objectives of this study were 1) to assess the time course of platelet adhesion in the small bowel after I/R and 2) to determine the roles of endothelial and/or platelet P selectin and P-selectin glycoprotein ligand-1 (PSGL-1) in this adhesion. The adhesion of fluorescently labeled platelets was monitored by intravital microscopy in postcapillary venules exposed to 45 min of ischemia and up to 8 h of reperfusion. Peak platelet adhesion was observed at 4 h of reperfusion. To assess the contributions of platelet and endothelial cell P-selectin, platelets from P-selectin-deficient and wild-type mice were infused into wild-type and P selectin-deficient mice, respectively. Platelets deficient in P-selectin exhibited low levels of adhesion comparable to that in sham-treated animals. In the absence of endothelial P-selectin, platelet adhesion was reduced by 65%. Treatment with a blocking antibody against PSGL-1 reduced adhesion by 57%. These results indicate that I/R induces a time-dependent platelet-endothelial adhesion response in postcapillary venules via a mechanism that involves PSGL-1 and both platelet and endothelial P-selectin, with platelet P-selectin playing a greater role. PMID- 12736151 TI - Enhanced capacitative calcium entry and TRPC channel gene expression in human LES smooth muscle. AB - Transient receptor potential channel (TRPC) genes encode Ca(2+)-permeable channels mediating capacitative Ca(2+) entry (CCE), which maintains intracellular Ca(2+) stores. We compared TRPC gene expression and CCE in human esophageal body (EB) and lower esophageal sphincter (LES), because these smooth muscles have distinct contractile functions that are likely associated with different Ca(2+) regulatory mechanisms. Circular layer smooth muscle cells were grown in primary culture. Transcriptional expression of TRPC genes was compared by semiquantitative RT-PCR. CCE was measured by fura 2 Ca(2+) fluorescence after blockade of sarcoplasmic reticulum Ca(2+)-ATPase with thapsigargin. mRNA for TRPC1, TRPC3, TRPC4, TRPC5, and TRPC6 was identified in EB and LES. TRPC3 and TRPC4 were more abundant in LES than EB. Basal concentration of free intracellular Ca(2+) ([Ca(2+)](i)) was similar in cells from LES (138 +/- 8 nmol/l) and EB (110 +/- 6 nmol/l) and increased with ACh (10 micromol/l; 650 +/- 28 and 590 +/- 21 nmol/l, respectively). With zero Ca(2+) in bath, thapsigargin (2 micromol/l) increased [Ca(2+)](i) more in LES (550 +/- 22 nmol/l) than EB (250 +/- 15 nmol/l, P < 0.001). Subsequent external application of 1 mmol/l Ca(2+) increased [Ca(2+)](i) more in LES (585 +/- 35 nmol/l) than EB (295 +/- 21 nmol/l, P < 0.001), indicating enhanced CCE in LES. This demonstrates CCE and TRPC transcriptional expression in human esophageal smooth muscle. In LES cells, enhanced CCE and expression of TRPC3 and TRPC4 may contribute to the physiological characteristics that distinguish LES from EB. PMID- 12736152 TI - Electrical stimulation reveals complex neuronal input and activation patterns in single myenteric guinea pig ganglia. AB - The myenteric plexus plays a key role in the control of gastrointestinal motility. We used confocal calcium imaging to study responses to electrical train stimulation (ETS) of interganglionic fiber tracts in entire myenteric ganglia of the guinea pig small intestine. ETS induced calcium transients in a subset of neurons: 52.2% responded to oral ETS, 65.4% to aboral ETS, and 71.7% to simultaneous oral and aboral ETS. A total of 41.3% of the neurons displayed convergence of oral and aboral ETS-induced responses. Responses could be reversibly blocked with TTX (10(-)6 M), demonstrating involvement of neuronal conduction, and by removal of extracellular calcium. omega-Conotoxin (5 x 10(-7) M) blocked the majority of responses and reduced the amplitude of residual responses by 45%, indicating the involvement of N-type calcium channels. Staining for calbindin and calretinin did not reveal different response patterns in these immunohistochemically identified neurons. We conclude that, at least for ETS close to a ganglion, confocal calcium imaging reveals complex oral and aboral input to individual myenteric neurons rather than a polarization in spread of activity. PMID- 12736153 TI - Identification of a basolateral Cl-/HCO3- exchanger specific to gastric parietal cells. AB - The basolateral Cl(-)/HCO(3)(-) exchanger in parietal cells plays an essential role in gastric acid secretion mediated via the apical gastric H(+)-K(+)-ATPase. Here, we report the identification of a new Cl(-)/HCO(3)(-) exchanger, which shows exclusive expression in mouse stomach and kidney, with expression in the stomach limited to the basolateral membrane of gastric parietal cells. Tissue distribution studies by RT-PCR and Northern hybridizations demonstrated the exclusive expression of this transporter, also known as SLC26A7, to stomach and kidney, with the stomach expression significantly more abundant. No expression was detected in the intestine. Cellular distribution studies by RT-PCR and Northern hybridizations demonstrated predominant localization of SLC26A7 in gastric parietal cells. Immunofluorescence labeling localized this exchanger exclusively to the basolateral membrane of gastric parietal cells, and functional studies in oocytes indicated that SLC26A7 is a DIDS-sensitive Cl(-)/HCO(3)(-) exchanger that is active in both acidic and alkaline pH(i). On the basis of its unique expression pattern and function, we propose that SLC26A7 is a basolateral Cl(-)/HCO(3)(-) exchanger in gastric parietal cells and plays a major role in gastric acid secretion. PMID- 12736154 TI - Role of nitric oxide in beta3-adrenoceptor activation on basal tone of internal anal sphincter. AB - Effects of activation of beta3-adrenoceptor (beta3-AR) have not been determined in the spontaneously tonic smooth muscle of the internal anal sphincter (IAS). The effects of disodium (R,R)-5-[2-[2-3-chlorophenyl)-2-hydroxyethyl] amino]propyl]-1,3-benzodioxole-2,2-dicarboxylate (CL 316243), a selective beta3 AR agonist, on the basal smooth muscle tone and direct release of nitric oxide (NO) by circular smooth muscle strips of the opossum IAS were determined. We also examined the presence of endothelial nitric oxide synthase (eNOS) protein by Western blot studies. CL 316243 produced a concentration-dependent relaxation of the smooth muscle that remained unmodified by different neurohumoral antagonists. The smooth muscle relaxation by CL 316243 was selectively antagonized by L 748337, a beta3-AR antagonist. Such relaxation was several times longer than by isoproterenol. The effect of CL 316243 was significantly attenuated by a nonselective NOS inhibitor N(omega)-nitro-l-arginine (l-NNA) and by putative inhibitor of eNOS l-N5-(1-iminoethyl)-ornithine dihydrochloride (l-NIO). Inhibitors of iNOS [N-(3-aminomethyl)benzyl acetamide 2HCl] and nNOS [1-[2 (trifluoromethylphenyl)imidazole]] had no effect on this relaxation. Relaxation of the IAS smooth muscle induced by CL 316243 was accompanied by an increased release of NO; this was attenuated by l-NNA and l-NIO. In addition, Western blot studies revealed the presence of eNOS in the circular smooth muscle of the IAS. These data demonstrate potent and protracted IAS smooth muscle relaxation by beta3-AR activation, which is partly transduced via NOS, possibly smooth muscle eNOS. Multiple signal-transduction pathways including NOS activation may explain the characteristic IAS relaxation by beta3-AR activation. The studies may have therapeutic implications in anorectal motility disorders. PMID- 12736155 TI - Vasodilator effects of L-arginine are stereospecific and augmented by insulin in humans. AB - The amino acid l-arginine, the precursor of nitric oxide (NO) synthesis, induces vasodilation in vivo, but the mechanism behind this effect is unclear. There is, however, some evidence to assume that the l-arginine membrane transport capacity is dependent on insulin plasma levels. We hypothesized that vasodilator effects of l-arginine may be dependent on insulin plasma levels. Accordingly, we performed two randomized, double-blind crossover studies in healthy male subjects. In protocol 1 (n = 15), subjects received an infusion of insulin (6 mU x kg(-1) x min(-1) for 120 min) or placebo and, during the last 30 min, l arginine or d-arginine (1 g/min for 30 min) x In protocol 2 (n = 8), subjects received l-arginine in stepwise increasing doses in the presence (1.5 mU x kg(-1) x min(-1)) or absence of insulin. Renal plasma flow and glomerular filtration rate were assessed by the para-aminohippurate and inulin plasma clearance methods, respectively. Pulsatile choroidal blood flow was assessed with laser interferometric measurement of fundus pulsation, and mean flow velocity in the ophthalmic artery was measured with Doppler sonography. l-arginine, but not d arginine, significantly increased renal and ocular hemodynamic parameters. Coinfusion of l-arginine with insulin caused a dose-dependent leftward shift of the vasodilator effect of l-arginine. This stereospecific renal and ocular vasodilator potency of l-arginine is enhanced by insulin, which may result from facilitated l-arginine membrane transport, enhanced intracellular NO formation, or increased NO bioavailability. PMID- 12736157 TI - Splanchnic free fatty acid kinetics. AB - These studies were conducted to assess the relationship between visceral adipose tissue free fatty acid (FFA) release and splanchnic FFA release. Steady-state splanchnic bed palmitate ([9,10-(3)H]palmitate) kinetics were determined from 14 sampling intervals from eight dogs with chronic indwelling arterial, portal vein, and hepatic vein catheters. We tested a model designed to predict the proportion of FFAs delivered to the liver from visceral fat by use of hepatic vein data. The model predicted that 15 +/- 2% of hepatic palmitate delivery originated from visceral lipolysis, which was greater (P = 0.004) than the 11 +/- 2% actually observed. There was a good relationship (r(2) = 0.63) between the predicted and observed hepatic palmitate delivery values, but the model overestimated visceral FFA release more at lower than at higher palmitate concentrations. The discrepancy could be due to differential uptake of FFAs arriving from the arterial vs. the portal vein or to release of FFAs in the hepatic circulatory bed. Splanchnic FFA release measured using hepatic vein samples was strongly related to visceral adipose tissue FFA release into the portal vein. This finding suggests that splanchnic FFA release is a good indicator of visceral adipose tissue lipolysis. PMID- 12736156 TI - Testosterone administration to men increases hepatic lipase activity and decreases HDL and LDL size in 3 wk. AB - Testosterone administration to men is known to decrease high-density lipoprotein cholesterol (HDL-C) and the subclasses HDL(2) and HDL(3). It also might increase the number of small, dense, low-density lipoprotein cholesterol (LDL-C) particles in hypogonadal men. The decrease in HDL-C and in LDL-C size is potentially mediated by hepatic lipase activity, which hydrolyzes lipoprotein phospholipids and triacylglycerol. To determine how HDL-C and LDL-C particles are affected by testosterone administration to eugonadal men, testosterone was administered as a supraphysiological dose (600 mg/wk) for 3 wk to elderly, obese, eugonadal men before elective hip or knee surgery, and lipids were measured by routine methods and by density gradient ultracentrifugation. Hepatic lipase activity increased >60% above baseline levels, and HDL-C, HDL(2), and HDL(3) significantly declined in 3 wk. In addition, the LDL-C peak particle density and the amount of LDL-C significantly increased. Testosterone is therefore a potent stimulator of hepatic lipase activity, decreasing HDL-C, HDL(2), and HDL(3) as well as increasing LDL particle density changes, all associated with increased cardiovascular risk. PMID- 12736158 TI - Effects of hyper- and hypothyroidism on thyroid hormone concentrations in regions of the rat brain. AB - The purpose of this study was to investigate the effects of hyper- and hypothyroidism on thyroid hormone concentrations and deiodinase activities in nine regions of the rat brain. Four weeks of treatment with 75 microg thyroxine (T4)/kg body wt induced a two- to threefold increase in T4 levels in all of these brain regions, whereas the 3,5,3'-triiodothyronine (T3) concentrations were reduced in five brain regions and remained unchanged in four. Even after 8 wk treatment with 300 microg T4/kg, the T3 concentrations remained normal in cortical areas, the hippocampus and amygdala, and were elevated only in areas in which inner-ring deiodinase activity was low or absent, and in the hypothalamus. At the subcellular level, nuclear concentrations of T3 were diminished in hypothyroidism but remained unaltered in hyperthyroidism in all areas except the hypothalamus, where they were enhanced. Cortical mitochondrial succinate dehydrogenase activity was reduced in both hypo- and hyperthyroidism in spite of normal T3 concentrations in hyperthyroid animals. The results show that nuclear T3 concentrations fall in hypothyroidism but do not change during severe hyperthyroidism in any brain region except the hypothalamus. Further research is thus needed to clarify the mechanisms mediating the numerous biochemical and psychological effects of hyperthyroidism. PMID- 12736159 TI - Regulation of leptin secretion from white adipocytes by free fatty acids. AB - Norepinephrine stimulates lipolysis and concurrently inhibits insulin-stimulated leptin secretion from white adipocytes. To assess whether there is a cause-effect relationship between these two metabolic events, the effects of fatty acids were investigated in isolated rat adipocytes incubated in buffer containing low (0.1%) and high (4%) albumin concentrations. Palmitic acid (1 mM) mimicked the inhibitory effects of norepinephrine (1 microM) on insulin (10 nM)-stimulated leptin secretion, but only at low albumin concentrations. Studies investigating the effects of the chain length of saturated fatty acids [from butyric (C4) to stearic (C18) acids] revealed that only fatty acids with a chain length superior or equal to eight carbons effectively inhibited insulin-stimulated leptin secretion. Long-chain mono- and polyunsaturated fatty acids constitutively present in adipocyte triglyceride stores (oleic, linoleic, gamma-linolenic, palmitoleic, eicosapentanoic, and docosahexanoic acids) also completely suppressed leptin secretion. Saturated and unsaturated fatty acids inhibited insulin-stimulated leptin secretion with the same potency and without any significant effect on basal secretion. On the other hand, inhibitors of mitochondrial fatty acid oxidation (palmoxirate, 2-bromopalmitate, 2 bromocaproate) attenuated the stimulatory effects of insulin on leptin release without reversing the effects of fatty acids or norepinephrine, suggesting that fatty acids do not need to be oxidized by the mitochondria to inhibit leptin release. These results demonstrate that long-chain fatty acids mimic the effects of norepinephrine on leptin secretion and suggest that they may play a regulatory role as messengers between stimulation of lipolysis by norepinephrine and inhibition of leptin secretion. PMID- 12736160 TI - Glucagon's actions are modified by the combination of epinephrine and gluconeogenic precursor infusion. AB - It was previously shown that glucagon and epinephrine have additive effects on both gluconeogenic and glycogenolytic flux. However, the changes in gluconeogenic substrates may have been limiting and thus may have prevented a synergistic effect on gluconeogenesis and a reciprocal inhibitory effect on glycogenolysis. Thus the aim of the present study was to determine if glucagon has a greater gluconeogenic and a smaller glycogenolytic effect in the presence of both epinephrine and clamped gluconeogenic precursors. Two groups (Epi and G + Epi + P) of 18-h-fasted conscious dogs were studied. In Epi, epinephrine was increased, and in G + Epi + P, glucagon and epinephrine were increased. Gluconeogenic precursors (lactate and alanine) were infused in G + Epi + P to match the rise that occurred in Epi. Insulin and glucose levels were also controlled and were similar in the two groups. Epinephrine and precursor administration increased glucagon's effect on gluconeogenesis (4.5-fold; P < 0.05) and decreased glucagon's effect on glycogenolysis (85%; P = 0.08). Thus, in the presence of both hormones, and when the gluconeogenic precursor supply is maintained, gluconeogenic flux is potentiated and glycogenolytic flux is inhibited. PMID- 12736161 TI - Regulation of adiponectin by adipose tissue-derived cytokines: in vivo and in vitro investigations in humans. AB - Adiponectin is an adipose tissue-specific protein that is abundantly present in the circulation and suggested to be involved in insulin sensitivity and development of atherosclerosis. Because cytokines are suggested to regulate adiponectin, the aim of the present study was to investigate the interaction between adiponectin and three adipose tissue-derived cytokines (IL-6, IL-8, and TNF-alpha). The study was divided into three substudies as follows: 1) plasma adiponectin and mRNA levels in adipose tissue biopsies from obese subjects [mean body mass index (BMI): 39.7 kg/m2, n = 6] before and after weight loss; 2) plasma adiponectin in obese men (mean BMI: 38.7 kg/m2, n = 19) compared with lean men (mean BMI: 23.4 kg/m2, n = 10) before and after weight loss; and 3) in vitro direct effects of IL-6, IL-8, and TNF-alpha on adiponectin mRNA levels in adipose tissue cultures. The results were that 1) weight loss resulted in a 51% (P < 0.05) increase in plasma adiponectin and a 45% (P < 0.05) increase in adipose tissue mRNA levels; 2) plasma adiponectin was 53% (P < 0.01) higher in lean compared with obese men, and plasma adiponectin was inversely correlated with adiposity, insulin sensitivity, and IL-6; and 3) TNF-alpha (P < 0.01) and IL-6 plus its soluble receptor (P < 0.05) decreased adiponectin mRNA levels in vitro. The inverse relationship between plasma adiponectin and cytokines in vivo and the cytokine-induced reduction in adiponectin mRNA in vitro suggests that endogenous cytokines may inhibit adiponectin. This could be of importance for the association between cytokines (e.g., IL-6) and insulin resistance and atherosclerosis. PMID- 12736162 TI - Pancreatic vasopressin V1b receptors: characterization in In-R1-G9 cells and localization in human pancreas. AB - Vasopressin (AVP) receptors present in In-R1-G9 cells, a hamster glucagon secreting alpha-pancreatic cell line, were characterized using SSR-149415, a selective nonpeptide V1b receptor antagonist, and reference AVP compounds. Binding experiments, using [3H]AVP as a ligand, identified a single population of high-affinity binding sites. SSR-149415 competitively inhibited this binding and exhibited nanomolar and stereospecific affinity for these sites. The affinity of various AVP/oxytocin ligands confirmed a V1b binding profile. In functional studies, AVP was a potent stimulant in inducing intracellular Ca2+ increase, glucagon secretion, and cell proliferation. These effects were fully antagonized by SSR-149415 with a nanomolar potency, whereas its diasteroisomer as well as two selective V1a and V2 receptor antagonists were much less potent. Additionally, the order of potency of AVP agonists and antagonists was in agreement with V1b mediated effects. By RT-PCR, we confirmed the presence of V1b receptor mRNA in both In-R1-G9 cells and in human pancreas. The distribution pattern of V1b receptors investigated in human pancreas by immunohistochemistry showed strong labeling in islets of Langerhans, and colocalization studies indicated that this receptor was expressed in alpha-glucagon, beta-insulin, and somatostatin pancreatic cells. Thus, in In-R1-G9 cells, AVP mediates intracellular Ca2+ increase, glucagon secretion, and cell proliferation by activating V1b receptors, and these effects are potently antagonized by SSR-149415. Moreover, the presence of V1b receptors also found in human Langerhans islets could suggest hormonal control of AVP in human pancreas. PMID- 12736163 TI - Bovine growth hormone-transgenic mice have major alterations in hepatic expression of metabolic genes. AB - Transgenic mice overexpressing growth hormone (GH) have been extensively used to study the chronic effects of elevated serum levels of GH. GH is known to have many acute effects in the liver, but little is known about the chronic effects of GH overexpression on hepatic gene expression. Therefore, we used DNA microarray to compare gene expression in livers from bovine GH (bGH)-transgenic mice and littermates. Hepatic expression of peroxisome proliferator-activated receptor alpha (PPARalpha) and genes involved in fatty acid activation, peroxisomal and mitochondrial beta-oxidation, and production of ketone bodies was decreased. In line with this expression profile, bGH-transgenic mice had a reduced ability to form ketone bodies in both the fed and fasted states. Although the bGH mice were hyperinsulinemic, the expression of sterol regulatory element-binding protein (SREBP)-1 and most lipogenic enzymes regulated by SREBP-1 was reduced, indicating that these mice are different from other insulin-resistant models with respect to expression of SREBP-1 and its downstream genes. This study also provides several candidate genes for the well-known association between elevated GH levels and cardiovascular disease, e.g., decreased expression of scavenger receptor class B type I, hepatic lipase, and serum paraoxonase and increased expression of serum amyloid A-3 protein. We conclude that bGH-transgenic mice display marked changes in hepatic genes coding for metabolic enzymes and suggest that GH directly or indirectly regulates many of these hepatic genes via decreased expression of PPARalpha and SREBP-1. PMID- 12736165 TI - Diabetic kidney disease in the db/db mouse. AB - Diabetic nephropathy is increasing in incidence and is now the number one cause of end-stage renal disease in the industrialized world. To gain insight into the genetic susceptibility and pathophysiology of diabetic nephropathy, an appropriate mouse model of diabetic nephropathy would be critical. A large number of mouse models of diabetes have been identified and their kidney disease characterized to various degrees. Perhaps the best characterized and most intensively investigated model is the db/db mouse. Because this model appears to exhibit the most consistent and robust increase in albuminuria and mesangial matrix expansion, it has been used as a model of progressive diabetic renal disease. In this review, we present the findings from various studies on the renal pathology of the db/db mouse model of diabetes in the context of human diabetic nephropathy. Furthermore, we discuss shortfalls of assessing functional renal disease in mouse models of diabetic kidney disease. PMID- 12736164 TI - Paradoxes of nitric oxide in the diabetic kidney. AB - As an important modulator of renal function and morphology, the nitric oxide (NO) system has been extensively studied in the diabetic kidney. However, a number of studies in different experimental and clinical settings have produced often confusing data and contradictory findings. We have reviewed a wide spectrum of findings and issues that have amassed concerning the pathophysiology of the renal NO system in diabetes, pointed out the controversies, and attempted to find some explanation for these discrepancies. Severe diabetes with profound insulinopenia can be viewed as a state of generalized NO deficiency, including in the kidney. However, we have focused our hypotheses and conclusions on the events occurring during moderate glycemic control with some degree of treatment with exogenous insulin, representing more the clinically applicable state of diabetic nephropathy. Available evidence suggests that diabetes triggers mechanisms that in parallel enhance and suppress NO bioavailability in the kidney. We hypothesize that during the early phases of nephropathy, the balance between these two opposing forces is shifted toward NO. This plays a role in the development of characteristic hemodynamic changes and may contribute to consequent structural alterations in glomeruli. Both endothelial (eNOS) and neuronal NO synthase can contribute to altered NO production. These enzymes, particularly eNOS, can be activated by Ca(2+)-independent and alternative routes of activation that may be elusive in traditional methods of investigation. As the duration of exposure to the diabetic milieu increases, factors that suppress NO bioavailability gradually prevail. Increasing accumulations of advanced glycation end products may be one of the culprits in this process. In addition, this balance is continuously modified by actual metabolic control and the degree of insulinopenia. PMID- 12736166 TI - Tubular reabsorption of myo-inositol vs. that of D-glucose in rat kidney in vivo et situ. AB - Filtered myo-inositol, an important renal intracellular organic osmolyte, is almost completely reabsorbed. To examine tubule sites and specificity and, thus possible mechanism of this reabsorption, we microinfused myo-[(3)H]inositol or D [(3)H]glucose into early proximal (EP), late proximal (LP), or early distal tubule sections of superficial nephrons and into long loops of Henle (LLH) of juxtamedullary nephrons and papillary vasa recta in rats in vivo et situ and determined urinary fractional recovery of the (3)H label compared with comicroinfused [(14)C]inulin. To determine the extent to which the proximal convoluted tubule (PCT) alone contributes to myo-inositol reabsorption, we also microperfused this tubule segment between EP and LP puncture sites. We examined specificity of reabsorptive carrier(s) by adding high concentrations of other polyols and monosaccharides to the infusate. The results show that >60% of the physiological glomerular load of myo-inositol can be reabsorbed in the PCT and >90% in the short loop of Henle (SLH) by a saturable, phloridzin-sensitive process. myo-Inositol can also be reabsorbed in the ascending limb of LLH and can move from papillary vasa recta blood into ipsilateral tubular structures. Essentially no reabsorption occurred in nephron segments beyond the SLH or in collecting ducts. Specificity studies indicate that reabsorption probably occurs via a luminal Na(+)-myo-inositol cotransporter. PMID- 12736167 TI - Nitric oxide synthesis influences the renal vascular response to heme oxygenase inhibition. AB - We studied the effects of the heme oxygenase (HO) inhibitor stannous mesoporphyrin (SnMP; 40 micromol/kg i.v.) on renal hemodynamics in anesthetized rats with and without 48-h pretreatment with N(G)-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide (NO) synthesis. SnMP decreased renal blood flow (RBF) and increased renal vascular resistance (RVR) in both groups. The SnMP induced reduction of RBF in L-NAME-pretreated rats was more prominent than in rats without pretreatment (43 +/- 7 vs. 13 +/- 3%) as was the SnMP-induced elevation of RVR (87 +/- 31 vs. 14 +/- 5%). The renal vasoconstrictor effect of SnMP is linked, in part, to amplification of prevailing neurohormonal constrictor mechanisms, since in L-NAME-pretreated rats it was prevented by concurrent administration of prazosin or losartan. However, SnMP (15 micromol/l) also elicits vasoconstriction in isolated, pressurized renal interlobular arteries and the response is more intense in vessels obtained from L-NAME-pretreated rats than from rats without pretreatment. These data indicate that the status of NO synthesis conditions the vascular response to HO inhibition in the rat kidney. PMID- 12736168 TI - Role of renal NO production in the regulation of medullary blood flow. AB - The unique role of nitric oxide (NO) in the regulation of renal medullary function is supported by the evidence summarized in this review. The impact of reduced production of NO within the renal medulla on the delivery of blood to the medulla and on the long-term regulation of sodium excretion and blood pressure is described. It is evident that medullary NO production serves as an important counterregulatory factor to buffer vasoconstrictor hormone-induced reduction of medullary blood flow and tissue oxygen levels. When NO synthase (NOS) activity is reduced within the renal medulla, either pharmacologically or genetically [Dahl salt-sensitive (S) rats], a super sensitivity to vasoconstrictors develops with ensuing hypertension. Reduced NO production may also result from reduced cellular uptake of l-arginine in the medullary tissue, resulting in hypertension. It is concluded that NO production in the renal medulla plays a very important role in sodium and water homeostasis and the long-term control of arterial pressure. PMID- 12736169 TI - Peptides that regulate food intake. PMID- 12736170 TI - The motivated hypothalamus. PMID- 12736171 TI - Inducing vasoconstriction by activating guanylyl cyclase. PMID- 12736172 TI - The spleen: another mystery about its function. PMID- 12736173 TI - Angiotensin II and control of sodium and water intake in the mouse. PMID- 12736174 TI - The long-lasting impact of postnatal neuropeptide Y. PMID- 12736175 TI - Peptides that regulate food intake: norepinephrine is not required for reduction of feeding induced by cholecystokinin. AB - CCK octapeptide (CCK-8) is released by the gut in response to a meal and acts via CCK(A) receptors on vagal afferents to induce satiety. However, the central neural pathways by which peripheral CCK-8 affects feeding are poorly understood. In the present study, we tested the hypothesis that norepinephrine (NE) is necessary for satiety induced by peripheral CCK-8 by using mice lacking dopamine beta-hydroxylase (Dbh(-/-)), the enzyme responsible for synthesizing NE and epinephrine from dopamine. We found that Dbh(-/-) mice are as responsive to the satiating effects of CCK-8 as their normal littermates. PMID- 12736176 TI - Peptides that regulate food intake: somatostatin alters intake of amino acid imbalanced diets and taste buds of tongue in rats. AB - The present studies were designed to evaluate a potential dose-dependent effect of somatostatin (SRIF) administered peripherally on intake of either a low protein basal diet or threonine-imbalanced diet (THR-IMB), on body weight gain (DeltaBW), gut motility, and on the histology of taste buds in rats. SRIF administration had a dual effect related to its concentration, increasing the intake of THR-IMB diet at low concentration and decreasing THR-IMB diet at high concentration. During the light phase, SRIF treatment increased the intake of THR IMB diet, suggesting that the usual anorectic effect induced by intake of THR-IMB diet was attenuated. High-dosage SRIF decreases gastrointestinal motility, which, in turn, can decrease food intake and DeltaBW. The combination of THR-IMB diet regimen and SRIF treatment also induced significant modifications on the taste buds of the tongue. The feeding response to an amino acid-imbalanced diet includes a learned aversion to the diet, and animals may use taste in establishing that aversion. Modifications of taste buds of SRIF-treated rats eating THR-IMB diet might explain the increase of imbalanced diet intake if treated rats perceive this food as less aversive. PMID- 12736177 TI - Peptides that regulate food intake: antagonism of opioid receptors reduces body fat in obese rats by decreasing food intake and stimulating lipid utilization. AB - Agonists to opioid receptors induce a positive energy balance, whereas antagonists at these receptors reduce food intake and body weight in rodent models of obesity. An analog of 3,4-dimethyl-4-(3-hydroxyphenyl)piperidine, LY255582, is a potent non-morphinan antagonist for mu-, kappa-, and delta receptors (K(i) of 0.4, 2.0, and 5.2 nM, respectively). In the present study, we examined the effects of oral LY255582 treatment on caloric intake, calorie expenditure, and body composition in dietary-induced obese rats. Acute oral treatment of LY255582 produced a dose-dependent decrease in energy intake and respiratory quotient (RQ), which correlated with the occupancy of central opioid receptors. Animals receiving chronic oral treatment with LY255582 for 14 days maintained a negative energy balance that was sustained by increased lipid use. Analysis of body composition revealed a reduction in fat mass accretion, with no change in lean body mass, in animals treated with LY255582. Therefore, chronic treatment with LY255582 reduces adipose tissue mass by reducing energy intake and stimulating lipid use. PMID- 12736178 TI - Peptides that regulate food intake: regional, metabolic, and circadian specificity of lateral hypothalamic orexin A feeding stimulation. AB - Orexin A (OX-A) administered in the lateral hypothalamus (LH) increases feeding in a dose-dependent manner. The LH is a relatively large neural structure with a heterogeneous profile of neural inputs, efferent projections, and orexin receptor distribution. We sought to determine the LH region most sensitive to the feeding stimulatory effect of OX-A injection. Fifty-six male Sprague-Dawley rats were fitted with cannulas 1 mm above four separate LH regions approximately 1 mm apart in the rostral-caudal direction. There were 14-16 animals/LH region. After recovery, animals received either artificial cerebrospinal fluid or OX-A (250, 500, or 1,000 pmol). To determine whether there is a circadian effect of LH OX-A on the feeding response, we performed injections at 0200, 0900, 1400, and 2100. Food intake was measured at 1, 2, and 4 h after injection. The most rostral extent of the LH was the only region in which injection of OX-A significantly stimulated feeding. Within this region, feeding was increased at all times of the day, although the most robust and only significant feeding response occurred after the afternoon injection (1400) of OX-A. To determine the extent to which the metabolic status of the rat contributed to the circadian specificity of orexin-induced feeding, animals were placed on a restricted diet and injected with OX-A in the most rostral region of the LH. Under these conditions, OX-A significantly increased feeding and more robustly when compared with animals on a nonrestricted diet. These data suggest that the rostral LH is the only region of the LH sensitive to the injection of OX-A, and the metabolic status of the animal at the time of injection may influence the feeding response to OX-A. PMID- 12736180 TI - Effects of fasting on thermoregulatory processes and the daily oscillations in rats. AB - To investigate the mechanism involved in the reduction of body core temperature (T(core)) during fasting in rats, which is selective in the light phase, we measured T(core), surface temperature, and oxygen consumption rate in fed control animals and in fasted animals on day 3 of fasting and day 4 of recovery at an ambient temperature (T(a)) of 23 degrees C by biotelemetry, infrared thermography, and indirect calorimetry, respectively. On the fasting day, 1) T(core) in the light phase decreased (P < 0.05) from the control; however, T(core) in the dark phase was unchanged, 2) tail temperature fell from the control (P < 0.05, from 30.7 +/- 0.1 to 23.9 +/- 0.1 degrees C in the dark phase and from 29.4 +/- 0.1 to 25.2 +/- 0.2 degrees C in the light phase), 3) oxygen consumption rate decreased from the control (P < 0.05, from 24.37 +/- 1.06 to 16.24 +/- 0.69 ml. min(-1). kg body wt(-0.75) in the dark phase and from 18.91 +/ 0.64 to 14.00 +/- 0.41 ml. min(-1). kg body wt(-0.75) in the light phase). All these values returned to the control levels on the recovery day. The results suggest that, in the fasting condition, T(core) in the dark phase was maintained by suppression of the heat loss mechanism, despite the reduction of metabolic heat production. In contrast, the response was weakened in the light phase, decreasing T(core) greatly. Moreover, the change in the regulation of tail blood flow was a likely mechanism to suppress heat loss. PMID- 12736179 TI - Peptides that regulate food intake: appetite-inducing accumbens manipulation activates hypothalamic orexin neurons and inhibits POMC neurons. AB - Corticolimbic circuits involving the prefrontal cortex, amygdala, and ventral striatum determine the reward value of food and might play a role in environmentally induced obesity. Chemical manipulation of the nucleus accumbens shell (AcbSh) has been shown to elicit robust feeding and Fos expression in the hypothalamus and other brain areas of satiated rats. To determine the neurochemical phenotype of hypothalamic neurons receiving input from the AcbSh, we carried out c-Fos/peptide double-labeling immunohistochemistry in various hypothalamic areas known to contain feeding peptides, from rats that exhibited a significant feeding response after AcbSh microinjection of the GABA(A) agonist muscimol. In the perifornical area, a significantly higher percentage of orexin neurons expressed Fos after muscimol compared with saline injection. In contrast, Fos expression was not induced in melanin-concentrating hormone and cocaine amphetamine-related transcript (CART) neurons. In the arcuate nucleus, Fos activation was significantly lower in neurons coexpressing CART and proopiomelanocortin, and there was a tendency for higher Fos expression in neuropeptide Y neurons. In the paraventricular nucleus, no significant activation of oxytocin and CART neurons was found. Thus AcbSh manipulation may elicit food intake through coordinated stimulation of hypothalamic neurons expressing orexigenic peptides and suppression of neurons expressing anorexigenic peptides. However, activation of many neurons not expressing these peptides suggests that additional peptides/transmitters in the lateral hypothalamus and accumbens projections to other brain areas might also be involved. PMID- 12736181 TI - Neuropeptides modulate rat chorda tympani responses. AB - The neuropeptide leptin has been shown to selectively modulate rat chorda tympani (CT) responses to sweet tastants. To explore whether other neuropeptides can modulate such responses, rat whole nerve CT responses to NaCl, HCl, quinine HCl, and sucrose were measured while administering cholecystokinin-8 (CCK-8), substance P(4-11) (SP(4-11)), or calcitonin gene-related peptide (CGRP). To avoid possible confounding effects on CT responses that take long times to develop, such as those that arise from intraperitoneal injections, we investigated the effects of the above peptides injected into the ipsilateral lingual artery (LA) on CT nerve responses during the initial seconds after a tastant was placed on the tongue. We found that CT responses to NaCl and HCl were increased by CCK-8 and decreased by CGRP. SP(4-11) had no noticeable effect. Peptide-induced CT responses to quinine HCl or sucrose were too small to accurately detect. These data suggest that at short latencies, after local infusion via the LA, neuropeptides can alter CT responses in a peptide-specific manner. PMID- 12736182 TI - Magnetic fields and the melatonin hypothesis: a study of workers chronically exposed to 50-Hz magnetic fields. AB - Because epidemiological studies report clinical disorders (mainly neurobehavioral alterations and/or cancer) that may be related to diminished melatonin secretion or to changes in its circadian rhythm in subjects living or working in environments exposed to magnetic fields, research on the effects of these fields in humans is particularly important. In this study, we examine the circadian rhythm of melatonin in 15 men exposed chronically and daily for a period of 1-20 yr, in the workplace and at home, to a 50-Hz magnetic field in search of any cumulative effect from those chronic conditions of exposure. The weekly geometric mean of individual exposures ranged from 0.1 to 2.6 microT. The results are compared with those for 15 unexposed men who served as controls (individual exposures ranged from 0.004 to 0.092 microT). Blood samples were taken hourly from 2000 to 0800. Nighttime urine was also collected and analyzed. This work shows that subjects exposed over a long period (up to 20 yr) and on a daily basis to magnetic fields experienced no changes in their plasma melatonin level, their urinary 6-sulfatoxymelatonin level, or the circadian rhythm of melatonin. Our data strongly suggest that magnetic fields do not have cumulative effects on melatonin secretion in humans and thus clearly rebut the "melatonin hypothesis" that a decrease in plasma melatonin concentration (or a disruption in its secretion) explains the occurrence of clinical disorders or cancers possibly related to magnetic fields. PMID- 12736183 TI - Expression of glycerokinase in brown adipose tissue is stimulated by the sympathetic nervous system. AB - The effect of cold exposure (4 degrees C) or prolonged norepinephrine infusion on the activity and mRNA levels of glycerokinase (GyK) was investigated in rat interscapular brown adipose tissue (BAT). Cold exposure for 12 and 24 h induced increases of 30% and 100%, respectively, in the activity of BAT GyK, which was paralleled by twofold and fourfold increase in enzyme mRNA levels. BAT hemidenervation resulted in reductions of 50% and 30% in GyK activity and in mRNA levels, respectively, in denervated pads from rats kept at 25 degrees C, and suppressed in these pads the cold-induced increases in both GyK activity and mRNA levels. The increase in GyK activity induced by cold exposure was not affected by phenoxybenzamine, but was markedly inhibited by previous administration of propranolol or actinomycin D. BAT GyK activity did not change significantly after 6 h of continuous subcutaneous infusion of norepinephrine (20 microg/h), but increased twofold and fourfold after 12 and 24 h, with no further increase after 72 h of infusion. Norepinephrine infusion also activated mRNA production, but the effect was comparatively smaller than that on enzyme activity. beta-Adrenergic agonists also stimulated GyK activity with the following relative magnitude of response: CL316243 (beta(3)) > isoproterenol (non-selective) > dobutamine (beta(1)). In vitro rates of incorporation of glycerol into glyceride-glycerol were increased in BAT from rats exposed to cold. The data suggest that in conditions of a sustained increase in BAT sympathetic flow there is a stimulation of GyK gene expression at the pretranslational level, with increased enzyme activity, mediated by beta-adrenoreceptors, mainly beta(3). PMID- 12736184 TI - Effect of portal hypertension on splenic blood flow, intrasplenic extravasation and systemic blood pressure. AB - We have previously shown that intrasplenic fluid extravasation is important in controlling blood volume. We proposed that, because the splenic vein flows in the portal vein, portal hypertension would increase splenic venous pressure and thus increase intrasplenic microvascular pressure and fluid extravasation. Given that the rat spleen has no capacity to store/release blood, intrasplenic fluid extravasation can be estimated by measuring the difference between splenic arterial inflow and venous outflow. In anesthetized rats, partial ligation of the portal vein rostral to the junction with the splenic vein caused portal venous pressure to rise from 4.5 +/- 0.5 to 12.0 +/- 0.9 mmHg (n = 6); there was no change in portal venous pressure downstream of the ligation, although blood flow in the liver fell. Splenic arterial flow did not change, but the arteriovenous flow differential increased from 0.8 +/- 0.3 to 1.2 +/- 0.1 ml/min (n = 6), and splenic venous hematocrit rose. Mean arterial pressure fell (101 +/- 5.5 to 95 +/ 4 mmHg). Splenic afferent nerve activity increased (5.6 +/- 0.9 to 16.2 +/- 0.7 spikes/s, n = 5). Contrary to our hypothesis, partial ligation of the portal vein caudal to the junction with the splenic vein (same increase in portal venous pressure but no increase in splenic venous pressure) also caused the splenic arteriovenous flow differential to increase (0.6 +/- 0.1 to 1.0 +/- 0.2 ml/min; n = 8). The increase in intrasplenic fluid efflux and the fall in mean arterial pressure after rostral portal vein ligation were abolished by splenic denervation. We propose there to be an intestinal/hepatic/splenic reflex pathway, through which is mediated the changes in intrasplenic extravasation and systemic blood pressure observed during portal hypertension. PMID- 12736185 TI - Activation of central melanocortin-4 receptor suppresses lipopolysaccharide induced fever in rats. AB - Activation of central melanocortin receptors (MCR) inhibits fever, but the identity of the MCR subtype(s) mediating this antipyretic effect is unknown. To determine whether selective central melanocortin receptor-4 (MC4R) activation produces antipyretic effects, the MC4R selective agonist MRLOB-0001 (CO-His-d-Phe Arg-Trp-Dab-NH(2)) was administered intracerebroventricularly to rats treated with Escherichia coli lipopolysaccharide (LPS, 30 microg/kg ip). Treatment with MRLOB-0001 (150 ng icv) did not lower core body temperature (T(c)) in afebrile rats but did suppress LPS-induced increases in T(c) and associated decreases in tail skin temperature (T(sk)), an indicator of vasomotor thermoeffector function. In contrast, systemic treatment with MRLOB-0001 (150 ng iv) did not produce similar antipyretic effects. Coadministration of the selective MC4R antagonist HS014 (1 microg icv) blocked the antipyretic effects of MRLOB-0001. HS014 alone (1 microg icv) had no significant effect on LPS-induced increases in T(c) or decreases in T(sk) and in afebrile rats had no significant effects on T(c) or T(sk). We conclude that pharmacological activation of central MC4R suppresses febrile increases in T(c) and that inhibition of heat conservation pathways may contribute to this effect. These findings suggest that the central MC4R may mediate the long-recognized antipyretic effects of centrally administered melanocortins. PMID- 12736186 TI - Mechanisms of bacterial lipopolysaccharide-induced endothelial apoptosis. AB - Gram-negative bacterial sepsis remains a common, life-threatening event. The prognosis for patients who develop sepsis-related complications, including the development of acute respiratory distress syndrome (ARDS), remains poor. A common finding among patients and experimental animals with sepsis and ARDS is endothelial injury and/or dysfunction. A component of the outer membrane of gram negative bacteria, lipopolysaccharide (LPS) or endotoxin, has been implicated in the pathogenesis of much of the endothelial cell injury and/or dysfunction associated with these disease states. LPS is a highly proinflammatory molecule that elicits a wide array of endothelial responses, including the upregulation of cytokines, adhesion molecules, and tissue factor. In addition to activation, LPS induces endothelial cell death that is apoptotic in nature. This review summarizes the evidence for LPS-induced vascular endothelial injury and examines the molecular signaling pathways that activate and inhibit LPS-induced endothelial apoptosis. Furthermore, the role of apoptotic signaling molecules in mediating LPS-induced activation of endothelial cells will be considered. PMID- 12736187 TI - Super-SOD: superoxide dismutase chimera fights off inflammation. PMID- 12736188 TI - Synthesis and anti-inflammatory activity of a chimeric recombinant superoxide dismutase: SOD2/3. AB - External surfaces of cells are normally protected by extracellular superoxide dismutase, SOD3, which binds to polyanions such as heparan sulfate. We constructed a fusion gene encoding a chimeric SOD consisting of the mature human mitochondrial SOD2 plus the COOH-terminal 26-amino acid heparin-binding "tail" from SOD3. This tail is responsible for the enzyme's affinity for endothelial surfaces. The fusion gene was expressed in Escherichia coli, and the fully active enzyme SOD2/3 was purified. Although native SOD2 has no affinity for heparin, SOD2/3 binds to a heparin-agarose column. In a rat model of acute lung injury induced by intratracheal instillation of IL-1, SOD2/3, SOD2, and denatured SOD2/3 showed 92%, 13.8%, and 0% reduction of lung leak, respectively. Only SOD2/3 prevented neutrophil accumulation. In the carrageenan-induced foot edema model in the rat, SOD2/3 reduced edema by 62% (P < 0.003) at a dose in which native SOD2 produced no significant effect. Thus SOD2/3 appears to have properties as a therapeutic anti-inflammatory agent that are greatly superior to other available forms of the enzyme. PMID- 12736189 TI - Glycogen availability does not affect the TCA cycle or TAN pools during prolonged, fatiguing exercise. AB - The hypothesis that fatigue during prolonged exercise arises from insufficient intramuscular glycogen, which limits tricarboxylic acid cycle (TCA) activity due to reduced TCA cycle intermediates (TCAI), was tested in this experiment. Seven endurance-trained men cycled at approximately 70% of peak O(2) uptake (Vo(2 peak)) until exhaustion with low (LG) or high (HG) preexercise intramuscular glycogen content. Muscle glycogen content was lower (P < 0.05) at fatigue than at rest in both trials. However, the increase in the sum of four measured TCAI (>70% of the total TCAI pool) from rest to 15 min of exercise was not different between trials, and TCAI content was similar after 103 +/- 15 min of exercise (2.62 +/- 0.31 and 2.59 +/- 0.28 mmol/kg dry wt for LG and HG, respectively), which was the point of volitional fatigue during LG. Subjects cycled for an additional 52 +/- 9 min during HG, and although glycogen was markedly reduced (P < 0.05) during this period, no further change in the TCAI pool was observed, thus demonstrating a clear dissociation between exercise duration and the size of the TCAI pool. Neither the total adenine nucleotide pool (TAN = ATP + ADP + AMP) nor IMP was altered compared with rest in either trial, whereas creatine phosphate levels were not different when values measured at fatigue were compared with those measured after 15 min of exercise. These data demonstrate that altered glycogen availability neither compromises TCAI pool expansion nor affects the TAN pool or creatine phosphate or IMP content during prolonged exercise to fatigue. Therefore, our data do not support the concept that a decrease in muscle TCAI during prolonged exercise in humans compromises aerobic energy provision or is the cause of fatigue. PMID- 12736190 TI - Different effects on human skeletal myosin heavy chain isoform expression: strength vs. combination training. AB - Myosin heavy chain (MHC) isoform expression changes with physical training. This may be one of the mechanisms for muscular adaptation to exercise. We aimed to investigate the effects of different strength-training protocols on MHC isoform expression, bearing in mind that alpha- MHC(slow) (newly identified MHC isoform) mRNA may be upregulated in response to training. Twelve volunteers performed a 6 wk strength training with maximum contractions (Max group), and another 12 of similar age performed combination training of maximum contractions and ballistic and stretch-shortening movements (Combi group). Muscle samples were taken from triceps brachii before and after training. MHC isoform composition was determined by SDS-PAGE silver staining, and mRNA levels of MHC isoforms were determined by RT-PCR. In Max group, there was an increase in MHC(2A) (49.4 to 66.7%, P < 0.01) and a decrease in MHC(2X) (33.4 to 19.5%, P < 0.01) after training, although there was no significant change in MHC(slow). In Combi group, there was also an increase in MHC(2A) (47.7 to 62.7%, P < 0.05) and a decrease in MHC(slow) (18.2 to 9.2%, P < 0.05) but no significant change in MHC(2X). An upregulation of alpha MHC(slow) mRNA was, therefore, found in both groups as a result of training. The strength training with maximum contractions led to a shift in MHC isoform composition from 2X to 2A, whereas the combined strength training produced an MHC isoform composition shift from slow to 2A. PMID- 12736191 TI - Altered myocardial Gs protein and adenylyl cyclase signaling in rats exposed to chronic hypoxia and normoxic recovery. AB - The present work has analyzed the consequences of chronic intermittent high altitude hypoxia for functioning of the G protein-mediated adenylyl cyclase (AC) signaling system in the right (RV) and left ventricular (LV) myocardium in rats. Adaptation to hypoxia did not appreciably affect the number of beta-adrenoceptors and the content of predominantly membrane-bound alpha-subunit (G(s)alpha) of the stimulatory G protein, but it raised the amount of cytosolic G(s)alpha in RV. The levels of myocardial inhibitory Galpha protein were not altered. Activity of AC stimulated by GTP, fluoride, forskolin, or isoprotertenol was reduced by approximately 50% in RV from chronically hypoxic rats, and a weaker depression was also found in LV. In addition, hypoxia significantly diminished a functional activity of membrane-bound G(s)alpha in both RV and LV. The RV baseline contractile function was markedly increased in chronically hypoxic animals, and its sensitivity to beta-adrenergic stimulation was decreased. Animals recovering from hypoxia for 5 wk still exhibited markedly elevated levels of cytosolic G(s)alpha and significantly lower activity of AC in RV than did age-matched controls, but contractile responsiveness to beta-agonists was normal. PMID- 12736192 TI - Genetic Models in Applied Physiology. Functional genomics in the mouse: powerful techniques for unraveling the basis of human development and disease. AB - Now that near-complete DNA sequences of both the mouse and human genomes are available, the next major challenge will be to determine how each of these genes functions, both alone and in combination with other genes in the genome. The mouse has a long and rich history in biological research, and many consider it a model organism for the study of human development and disease. Over the past few years, exciting progress has been made in developing techniques for chromosome engineering, mutagenesis, mapping and maintenance of mutations, and identification of mutant genes in the mouse. In this mini-review, many of these powerful techniques will be presented along with their application to the study of development, physiology, and disease. PMID- 12736193 TI - Genetic Models in Applied Physiology. HXB/BXH rat recombinant inbred strain platform: a newly enhanced tool for cardiovascular, behavioral, and developmental genetics and genomics. AB - This review deals with the largest set of rat recombinant inbred (RI) strains and summarizes past and recent accomplishments with this platform for genetic mapping and analyses of divergent and complex traits. This strain, derived by crossing the spontaneously hypertensive rat, SHR/Ola, with a Brown Norway congenic, BN-Lx, carrying polydactyly-luxate syndrome, is referred to as HXB/BXH. The RI strain set has been used for linkage and association studies to identify quantitative trait loci for numerous cardiovascular phenotypes, including arterial pressure, stress-elicited heart rate, and pressor response, and metabolic traits, including insulin resistance, dyslipidemia and glucose handling, and left ventricular hypertrophy. The strain's utility has been enhanced with development of a new framework marker-based map and strain distribution patterns of polymorphic markers. Quantitative trait loci for behavioral traits mapped include loci for startle motor response and habituation, anxiety and locomotion traits associated with elevated plus maze, and conditioned taste aversion. The polydactyly-luxate syndrome Lx mutation has allowed the study of alleles important to limb development and malformation phenotypes as well as teratogens. The RI strains have guided development of numerous congenic strains to test locus assignments and to study the effect of genetic background. Although these strains were originally developed to aid in studies of rat genetic hypertension and morphogenetic abnormalities, this rodent platform has been shown to be equally powerful for a wide spectrum of traits and endophenotypes. These strains provide a ready and available vehicle for many physiological and pharmacological studies. PMID- 12736195 TI - Genetic Models in Applied Physiology. Merosin deficiency leads to alterations in passive and active skeletal muscle mechanics. AB - The role of extracellular elements on the mechanical properties of skeletal muscles is unknown. Merosin is an essential extracellular matrix protein that forms a mechanical junction between the sarcolemma and collagen. Therefore, it is possible that merosin plays a role in force transmission between muscle fibers and collagen. We hypothesized that deficiency in merosin may alter passive muscle stiffness, viscoelastic properties, and contractile muscle force in skeletal muscles. We used the dy/dy mouse, a merosin-deficient mouse model, to examine changes in passive and active muscle mechanics. After mice were anesthetized and the diaphragm or the biceps femoris hindlimb muscle was excised, passive length tension relationships, stress-relaxation curves, or isometric contractile properties were determined with an in vitro biaxial mechanical testing apparatus. Compared with controls, extensibility was smaller in the muscle fiber direction and the transverse fiber direction of the mutant mice. The relaxed elastic modulus was smaller in merosin-deficient diaphragms compared with controls. Interestingly, maximal muscle tetanic stress was depressed in muscles from the mutant mice during uniaxial loading but not during biaxial loading. However, presence of transverse passive stretch increases maximal contractile stress in both the mutant and normal mice. Our data suggest that merosin contributes to muscle passive stiffness, viscoelasticity, and contractility and that the mechanism by which force is transmitted between adjacent myofibers via merosin possibly in shear. PMID- 12736196 TI - Airway surface fluid desiccation during isocapnic hyperpnea. PMID- 12736197 TI - Interpreting VO2 kinetics in heavy exercise revisited. PMID- 12736198 TI - The contribution of N-glycans and their processing in the endoplasmic reticulum to glycoprotein biosynthesis. AB - The attachment of N-glycans to nascent glycoproteins in the endoplasmic reticulum (ER) is intimately related to glycoprotein biogenesis. Processing of N-linked oligosaccharides begins in the ER and participates in glycoprotein folding and assembly. The elucidation of N-glycan processing mechanisms in the ER is uncovering their role in glycoprotein biosynthesis. PMID- 12736199 TI - Dystroglycan glycosylation and its role in matrix binding in skeletal muscle. AB - Dystroglycan is an essential component of the dystrophin-glycoprotein complex. Three glycan sequencing studies have identified O-linked mannose chains, including NeuAcalpha 2,3Galbeta 1,4GlcNAcbeta 1,2Manalpha-O, on alpha dystroglycan. Chemical deglycosylation of alpha dystroglycan, antibody blocking studies, and glycan blocking studies all suggest that the O-linked glycans on alpha dystroglycan mediate the binding of extracellular matrix proteins in skeletal muscle. Structural data on laminin G domains and agrin-binding studies also suggest this is the case. Dystroglycan, however, is able to bind proteins via mechanisms that do not involve O-linked glycans. Moreover, laminin and other matrix proteins can bind cell adhesion molecules via their glycan chains. Thus although complex and sometimes not overly convincing, these data suggest that glycosylation plays an important role in dystroglycan binding and function in skeletal muscle. PMID- 12736201 TI - Arabidopsis MSI1 is required for epigenetic maintenance of reproductive development. AB - WD40 repeat proteins similar to yeast MSI1 are conserved in animals and plants, in which they participate in complexes involved in chromatin metabolism. Although MSI1-like proteins are well characterised biochemically, their function in the development of multicellular eukaryotes is not well understood. We constructed Arabidopsis plants in which the AtMSI1 protein level was altered. Strong ectopic expression of AtMSI1 produced no visible altered phenotype, but reduction of AtMSI1 dramatically affected development. The primary shoot apical meristem was unable to develop organs after the transition to flowering. Flowers that developed on floral shoots from axillary meristems experienced a progressive loss of floral morphology, including a reduction in size of the petals and stamens and the development of carpel-like sepals. Ovule development was disrupted in all flowers, resulting in complete female sterility. Molecular analysis of the mutant plants revealed that AtMSI1 is required to maintain the correct temporal and organ-specific expression of homeotic genes, including AGAMOUS and APETALA2. In contrast, FAS1 and FAS2, which together with AtMSI1 form the chromatin assembly complex CAF-1, are not required for repression of these genes. Therefore, AtMSI1 has specific functions in addition to CAF-1-mediated chromatin assembly. Efficient formation of heterochromatin, but not methylation of centromeric DNA repeats, depends on AtMSI1 presence demonstrating a key role of AtMSI1 in maintenance of chromatin structure. PMID- 12736200 TI - Glycobiology of neuromuscular disorders. AB - There has been a recent explosion in the identification of neuromuscular diseases caused by mutations in genes that affect carbohydrate metabolism or protein glycosylation. A number of these findings relate to defects in the glycosylation of alpha dystroglycan. Alpha dystroglycan is an essential component of the dystrophin-glycoprotein complex, and aberrant glycosylation of alpha dystroglycan is associated with multiple forms of muscular dystrophy in mice and humans. We review the evidence that defects in dystroglycan glycosylation cause muscular dystrophy. In addition, we review evidence that glycobiology is important in other disorders that affect muscle, including hereditary inclusion body myopathy type II and congenital disorders of glycosylation. Finally, we discuss the long term potential of glycotherapies for muscle disorders. PMID- 12736202 TI - The C. elegans G-protein-coupled receptor SRA-13 inhibits RAS/MAPK signalling during olfaction and vulval development. AB - In C. elegans, the RAS/MAPK pathway is used in different tissues to regulate various cell fate decisions. Several positive and negative regulators tightly control the activity of the RAS/MAPK pathway at different steps. We demonstrate a link between a G-protein-coupled receptor signalling pathway and the RAS/MAPK cascade. SRA-13, a member of the SRA family of chemosensory receptors, negatively regulates RAS/MAPK signalling during vulval induction and the olfaction of volatile attractants. Epistasis analysis indicates that SRA-13 inhibits the RAS/MAPK pathway at the level or upstream of MAPK. In both tissues, the vulval precursor cells and the chemosensory neurones, SRA-13 acts through the GPA-5 Galpha protein subunit, suggesting a common mechanism of crosstalk. Moreover, we find that vulval induction is repressed by food withdrawal during larval development and that SRA-13 activity is required for the suppression of vulval induction in response to food starvation. Thus, SRA-13 may serve to adapt the activity of the RAS/MAPK pathway to environmental conditions. PMID- 12736203 TI - Clonal expansion of ovarian germline stem cells during niche formation in Drosophila. AB - Stem cell niches are specific regulatory microenvironments formed by neighboring stromal cells. Owing to difficulties in identifying stem cells and their niches in many systems, mechanisms that control niche formation and stem cell recruitment remain elusive. In the Drosophila ovary, two or three germline stem cells (GSCs) have recently been shown to reside in a niche, in which terminal filaments (TFs) and cap cells are two major components. We report that signals from newly formed niches promote clonal expansion of GSCs during niche formation in the Drosophila ovary. After the formation of TFs and cap cells, anterior primordial germ cells (PGCs) adjacent to TFs/cap cells can develop into GSCs at the early pupal stage while the rest directly differentiate. The anterior PGCs are very mitotically active and exhibit two division patterns with respect to cap cells. One of these patterns generates two daughters that both contact cap cells and potentially become GSCs. Our lineage tracing study confirms that one PGC can generate two or three GSCs to occupy a whole niche ('clonal expansion'). decapentaplegic (dpp), the Drosophila homolog of human bone morphogenetic protein 2/4, is expressed in anterior somatic cells of the gonad, including TFs/cap cells. dpp overexpression promotes PGC proliferation and causes the accumulation of more PGCs in the gonad. A single PGC mutant for thick veins, encoding an essential dpp receptor, loses the ability to clonally populate a niche. Therefore, dpp is probably one of the mitotic signals that promote the clonal expansion of GSCs in a niche. This study also suggests that signals from newly formed niche cells are important for expanding stem cells and populating niches. PMID- 12736204 TI - The C. elegans LIM homeobox gene lin-11 specifies multiple cell fates during vulval development. AB - LIM homeobox family members regulate a variety of cell fate choices during animal development. In C. elegans, mutations in the LIM homeobox gene lin-11 have previously been shown to alter the cell division pattern of a subset of the 2 degrees lineage vulval cells. We demonstrate multiple functions of lin-11 during vulval development. We examined the fate of vulval cells in lin-11 mutant animals using five cellular markers and found that lin-11 is necessary for the patterning of both 1 degrees and 2 degrees lineage cells. In the absence of lin-11 function, vulval cells fail to acquire correct identity and inappropriately fuse with each other. The expression pattern of lin-11 reveals dynamic changes during development. Using a temporally controlled overexpression system, we show that lin-11 is initially required in vulval cells for establishing the correct invagination pattern. This process involves asymmetric expression of lin-11 in the 2 degrees lineage cells. Using a conditional RNAi approach, we show that lin 11 regulates vulval morphogenesis. Finally, we show that LDB-1, a NLI/Ldb1/CLIM2 family member, interacts physically with LIN-11, and is necessary for vulval morphogenesis. Together, these findings demonstrate that temporal regulation of lin-11 is crucial for the wild-type vulval patterning. PMID- 12736205 TI - Development of the Drosophila mushroom bodies: elaboration, remodeling and spatial organization of dendrites in the calyx. AB - One Drosophila mushroom body (MB) is derived from four indistinguishable cell lineages, development of which involves sequential generation of multiple distinct types of neurons. Differential labeling of distinct MB clones reveals that MB dendrites of different clonal origins are well mixed at the larval stage but become restricted to distinct spaces in adults. Interestingly, a small dendritic domain in the adult MB calyx remains as a fourfold structure that, similar to the entire larval calyx, receives dendritic inputs from all four MB clones. Mosaic analysis of single neurons demonstrates that MB neurons, which are born around pupal formation, acquire unique dendritic branching patterns and consistently project their primary dendrites into the fourfold dendritic domain. Distinct dendrite distribution patterns are also observed for other subtypes of MB neurons. In addition, pruning of larval dendrites during metamorphosis allows for establishment of adult-specific dendrite elaboration/distribution patterns. Taken together, subregional differences exist in the adult Drosophila MB calyx, where processing and integration of distinct types of sensory information begin. PMID- 12736206 TI - Analysis of PINCH function in Drosophila demonstrates its requirement in integrin dependent cellular processes. AB - Integrins play a crucial role in cell motility, cell proliferation and cell survival. The evolutionarily conserved LIM protein PINCH is postulated to act as part of an integrin-dependent signaling complex. In order to evaluate the role of PINCH in integrin-mediated cellular events, we have tested directly the in vivo function of PINCH in Drosophila melanogaster. We demonstrate that the steamer duck (stck) alleles that were first identified in a screen for potential integrin effectors represent mutations in Drosophila pinch. stck mutants die during embryogenesis, revealing a key role for PINCH in development. Muscle cells within embryos that have compromised PINCH function display disturbed actin organization and cell-substratum adhesion. Mutation of stck also causes failure of integrin dependent epithelial cell adhesion in the wing. Consistent with the idea that PINCH could contribute to integrin function, PINCH protein colocalizes with betaPS integrin at sites of actin filament anchorage in both muscle and wing epithelial cells. Furthermore, we show that integrins are required for proper localization of PINCH at the myotendinous junction. The integrin-linked kinase, ILK, is also essential for integrin function. We demonstrate that Drosophila PINCH and ILK are complexed in vivo and are coincident at the integrin-rich muscle-attachment sites in embryonic muscle. Interestingly, ILK localizes appropriately in stck mutant embryos, therefore the phenotypes exhibited by the stck mutants are not attributable to mislocalization of ILK. Our results provide direct genetic evidence that PINCH is essential for Drosophila development and is required for integrin-dependent cell adhesion. PMID- 12736207 TI - Translational repression of a C. elegans Notch mRNA by the STAR/KH domain protein GLD-1. AB - In C. elegans, the Notch receptor GLP-1 is localized within the germline and early embryo by translational control of glp-1 mRNA. RNA elements in the glp-1 3'untranslated region (3' UTR) are necessary for repression of glp-1 translation in germ cells, and for localization of translation to anterior cells of the early embryo. The direct regulators of glp-1 mRNA are not known. Here, we show that a 34 nucleotide region of the glp-1 3' UTR contains two regulatory elements, an element that represses translation in germ cells and posterior cells of the early embryo, and an element that inhibits repressor activity to promote translation in the embryo. Furthermore, we show that the STAR/KH domain protein GLD-1 binds directly and specifically to the repressor element. Depletion of GLD-1 activity by RNA interference causes loss of endogenous glp-1 mRNA repression in early meiotic germ cells, and in posterior cells of the early embryo. Therefore, GLD-1 is a direct repressor of glp-1 translation at two developmental stages. These results suggest a new function for GLD-1 in regulating early embryonic asymmetry. Furthermore, these observations indicate that precise control of GLD-1 activity by other regulatory factors is important to localize this Notch receptor, and contributes to the spatial organization of Notch signaling. PMID- 12736208 TI - The isthmic organizer signal FGF8 is required for cell survival in the prospective midbrain and cerebellum. AB - Numerous studies have demonstrated that the midbrain and cerebellum develop from a region of the early neural tube comprising two distinct territories known as the mesencephalon (mes) and rostral metencephalon (met; rhombomere 1), respectively. Development of the mes and met is thought to be regulated by molecules produced by a signaling center, termed the isthmic organizer (IsO), which is localized at the boundary between them. FGF8 and WNT1 have been implicated as key components of IsO signaling activity, and previous studies have shown that in Wnt1(-/-) embryos, the mes/met is deleted by the 30 somite stage ( approximately E10) (McMahon, A. P. and Bradley, A. (1990) Cell 62, 1073-1085). We have studied the function of FGF8 in mouse mes/met development using a conditional gene inactivation approach. In our mutant embryos, Fgf8 expression was transiently detected, but then was eliminated in the mes/met by the 10 somite stage ( approximately E8.75). This resulted in a failure to maintain expression of Wnt1 as well as Fgf17, Fgf18, and Gbx2 in the mes/met at early somite stages, and in the absence of the midbrain and cerebellum at E17.5. We show that a major cause of the deletion of these structures is ectopic cell death in the mes/met between the 7 and 30 somite stages. Interestingly, we found that the prospective midbrain was deleted at an earlier stage than the prospective cerebellum. We observed a remarkably similar pattern of cell death in Wnt1 null homozygotes, and also detected ectopic mes/met cell death in En1 null homozygotes. Our data show that Fgf8 is part of a complex gene regulatory network that is essential for cell survival in the mes/met. PMID- 12736209 TI - Targeted ablation of CCAP neuropeptide-containing neurons of Drosophila causes specific defects in execution and circadian timing of ecdysis behavior. AB - Insect growth and metamorphosis is punctuated by molts, during which a new cuticle is produced. Every molt culminates in ecdysis, the shedding of the remains of the old cuticle. Both the timing of ecdysis relative to the molt and the actual execution of this vital insect behavior are under peptidergic neuronal control. Based on studies in the moth, Manduca sexta, it has been postulated that the neuropeptide Crustacean cardioactive peptide (CCAP) plays a key role in the initiation of the ecdysis motor program. We have used Drosophila bearing targeted ablations of CCAP neurons (CCAP KO animals) to investigate the role of CCAP in the execution and circadian regulation of ecdysis. CCAP KO animals showed specific defects at ecdysis, yet the severity and nature of the defects varied at different developmental stages. The majority of CCAP KO animals died at the pupal stage from the failure of pupal ecdysis, whereas larval ecdysis and adult eclosion behaviors showed only subtle defects. Interestingly, the most severe failure seen at eclosion appeared to be in a function required for abdominal inflation, which could be cardioactive in nature. Although CCAP KO populations exhibited circadian eclosion rhythms, the daily distribution of eclosion events (i.e., gating) was abnormal. Effects on the execution of ecdysis and its circadian regulation indicate that CCAP is a key regulator of the behavior. Nevertheless, an unexpected finding of this work is that the primary functions of CCAP as well as its importance in the control of ecdysis behaviors may change during the postembryonic development of Drosophila. PMID- 12736210 TI - Oriented cell divisions asymmetrically segregate aPKC and generate cell fate diversity in the early Xenopus embryo. AB - A key feature of early vertebrate development is the formation of superficial, epithelial cells that overlie non-epithelial deep cells. In Xenopus, deep and superficial cells show a range of differences, including a different competence for primary neurogenesis. We show that the two cell populations are generated during the blastula stages by perpendicularly oriented divisions. These take place during several cell divisions, in a variable pattern, but at a percentage that varies little between embryos and from one division to the next. The orientation of division correlates with cell shape suggesting that simple geometric rules may control the orientation of division in this system. We show that dividing cells are molecularly polarised such that aPKC is localised to the external, apical, membrane. Membrane localised aPKC can be seen as early as the one-cell stage and during the blastula divisions, it is preferentially inherited by superficial cells. Finally, we show that when 64-cell stage isolated blastomeres divide perpendicularly and the daughters are cultured separately, only the progeny of the cells that inherit the apical membrane turn on the bHLH gene, ESR6e. We conclude that oriented cell divisions generate the superficial and deep cells and establish cell fate diversity between them. PMID- 12736212 TI - Loss of Tbx4 blocks hindlimb development and affects vascularization and fusion of the allantois. AB - Tbx4 is a member of the T-box family of transcription factor genes, which have been shown to play important roles in development. We have ablated Tbx4 function using targeted mutagenesis in the mouse. Embryos homozygous for the null allele fail to undergo chorioallantoic fusion and die by 10.5 days post coitus. The allantoises of Tbx4-mutant embryos are stunted, apoptotic and display abnormal differentiation. Endothelial cells within mutant allantoises do not undergo vascular remodeling. Heterozygous embryos show a mild, transient growth defect in the allantois. Induction of a hindlimb field occurs normally in Tbx4 mutants and initial patterning of the hindlimb bud appears normal. However, hindlimb buds from Tbx4 mutants fail to develop either in vivo or in vitro and do not maintain Fgf10 expression in the mesenchyme. The expression of another, closely-linked, T box gene, Tbx2, is reduced in both the hindlimb and the allantois of Tbx4-mutant embryos prior to the development of overt morphological abnormalities, which suggests that Tbx4 regulates Tbx2 in these tissues. PMID- 12736211 TI - The zebrafish forkhead transcription factor Foxi1 specifies epibranchial placode derived sensory neurons. AB - Vertebrate epibranchial placodes give rise to visceral sensory neurons that transmit vital information such as heart rate, blood pressure and visceral distension. Despite the pivotal roles they play, the molecular program underlying their development is not well understood. Here we report that the zebrafish mutation no soul, in which epibranchial placodes are defective, disrupts the fork headrelated, winged helix domain-containing protein Foxi1. Foxi1 is expressed in lateral placodal progenitor cells. In the absence of foxi1 activity, progenitor cells fail to express the basic helix-loop-helix gene neurogenin that is essential for the formation of neuronal precursors, and the paired homeodomain containing gene phox2a that is essential for neuronal differentiation and maintenance. Consequently, increased cell death is detected indicating that the placodal progenitor cells take on an apoptotic pathway. Furthermore, ectopic expression of foxi1 is sufficient to induce phox2a-positive and neurogenin positive cells. Taken together, these findings suggest that Foxi1 is an important determination factor for epibranchial placodal progenitor cells to acquire both neuronal fate and subtype visceral sensory identity. PMID- 12736213 TI - The Xenopus LIM-homeodomain protein Xlim5 regulates the differential adhesion properties of early ectoderm cells. AB - One of the earliest lineage restriction events in embryogenesis is the specification of the primary germ layers: ectoderm, mesoderm and endoderm. In Xenopus, germ layer specification occurs prior to gastrulation and requires the transcription factor VegT both for the cell-autonomous specification of endoderm and the generation of mesoderm-inducing signals. In the absence of VegT, ectoderm is formed in all regions of the embryo. In this work, we show that VegT-depleted vegetal cells (prospective endoderm) behave like animal cells in sorting assays and ectopically express early markers of ectoderm. To gain insight into how ectoderm is specified, we looked for candidate ectoderm-specific genes that are ectopically expressed in VegT-depleted embryos, and examined the role of one of these, the LIM homeobox gene Xlim5, in ectoderm development. We show that overexpression of Xlim5 in prospective endoderm cells is sufficient to impair sorting of animal cells from vegetal cells but is not sufficient (at similar doses) to induce ectoderm-specific genes. In whole embryos, Xlim5 causes vegetal cells to segregate inappropriately to other germ layers and express late differentiation markers of that germ layer. Inhibition of Xlim5 function using an Engrailed repressor construct or a morpholino oligonucleotide causes loss of animal cell adhesion or delay in neural fold morphogenesis, respectively, without significantly affecting early ectoderm gene expression. Taken together, our results provide evidence that a primary role for Xlim5 is to specifically regulate differential cell adhesion behaviour of the ectoderm. PMID- 12736215 TI - Multiple levels of transcriptional and post-transcriptional regulation are required to define the domain of Hoxb4 expression. AB - Hox genes are key determinants of anteroposterior patterning of animal embryos, and spatially restricted expression of these genes is crucial to this function. In this study, we demonstrate that expression of Hoxb4 in the paraxial mesoderm of the mouse embryo is transcriptionally regulated in several distinct phases, and that multiple regulatory elements interact to maintain the complete expression domain throughout embryonic development. An enhancer located within the intron of the gene (region C) is sufficient for appropriate temporal activation of expression and the establishment of the correct anterior boundary in the paraxial mesoderm (somite 6/7). However, the Hoxb4 promoter is required to maintain this expression beyond 8.5 dpc. In addition, sequences within the 3' untranslated region (region B) are necessary specifically to maintain expression in somite 7 from 9.0 dpc onwards. Neither the promoter nor region B can direct somitic expression independently, indicating that the interaction of regulatory elements is crucial for the maintenance of the paraxial mesoderm domain of Hoxb4 expression. We further report that the domain of Hoxb4 expression is restricted by regulating transcript stability in the paraxial mesoderm and by selective translation and/or degradation of protein in the neural tube. Moreover, the absence of Hoxb4 3'-untranslated sequences from transgene transcripts leads to inappropriate expression of some Hoxb4 transgenes in posterior somites, indicating that there are sequences within region B that are important for both transcriptional and post-transcriptional regulation. PMID- 12736214 TI - Ogon/Secreted Frizzled functions as a negative feedback regulator of Bmp signaling. AB - The zebrafish mutant ogon (also called mercedes and short tail) displays ventralized phenotypes similar to the chordino (dino) mutant, in which the gene for the Bmp antagonist Chordin is mutated. We isolated the gene responsible for ogon by a positional cloning strategy and found that the ogon locus encodes a zebrafish homolog of Secreted Frizzled (Sizzled), which has sequence similarity to a Wnt receptor, Frizzled. Unlike other secreted Frizzled-related proteins (sFrps) and the Wnt inhibitor Dickkopf1, the misexpression of Ogon/Sizzled dorsalized, but did not anteriorize, the embryos, suggesting a role for Ogon/Sizzled in Bmp inhibition. Ogon/Sizzled did not inhibit a Wnt8-dependent transcription in the zebrafish embryo. ogon/sizzled was expressed on the ventral side from the late blastula through the gastrula stages. The ventral ogon/sizzled expression in the gastrula stage was reduced or absent in the swirl/bmp2b mutants but expanded in the chordino mutants. Misexpression of ogon/sizzled did not dorsalize the chordino mutants, suggesting that Ogon/Sizzled required Chordin protein for dorsalization and Bmp inhibition. These data indicate that Ogon/Sizzled functions as a negative regulator of Bmp signaling and reveal a novel role for a sFrp in dorsoventral patterning. PMID- 12736216 TI - Tbx5 and Tbx4 trigger limb initiation through activation of the Wnt/Fgf signaling cascade. AB - A tight loop between members of the fibroblast growth factor and the Wnt families plays a key role in the initiation of vertebrate limb development. We show for the first time that Tbx5 and Tbx4 are directly involved in this process. When dominant-negative forms of these Tbx genes were misexpressed in the chick prospective limb fields, a limbless phenotype arose with repression of both Wnt and Fgf genes By contrast, when Tbx5 and Tbx4 were misexpressed in the flank, an additional wing-like and an additional leg-like limbs were induced, respectively. This additional limb formation was accompanied by the induction of both Wnt and Fgf genes These results highlight the pivotal roles of Tbx5 and Tbx4 during limb initiation, specification of forelimb/hindlimb and evolution of tetrapod limbs, placing Tbx genes at the center of a highly conserved genetic program. PMID- 12736217 TI - Tbx5 is required for forelimb bud formation and continued outgrowth. AB - Tbx5 is a T-box transcription factor expressed exclusively in the developing forelimb but not in the developing hindlimb of vertebrates. Tbx5 is first detected in the prospective forelimb mesenchyme prior to overt limb bud outgrowth and its expression is maintained throughout later limb development stages. Direct evidence for a role of Tbx5 in forelimb development was provided by the discovery that mutations in human TBX5 cause Holt-Oram Syndrome (HOS), a dominant disorder characterised predominantly by upper(fore) limb defects and heart abnormalities. Misexpression studies in the chick have demonstrated a role for this gene in limb type specification. Using a conditional knockout strategy in the mouse to delete Tbx5 gene function in the developing forelimb, we demonstrate that this gene is also required at early limb bud stages for forelimb bud development. In addition, by misexpressing dominant-negative and dominant-activated forms of Tbx5 in the chick wing we provide evidence that this gene is also required at later stages of limb bud development for continued limb outgrowth. Our results provide a context to understand the defects observed in HOS caused by haploinsufficiency of TBX5 in human. Moreover, our results also demonstrate that limb bud outgrowth and specification of limb identity are linked by a requirement for Tbx5. PMID- 12736218 TI - Elevated SMAD1/beta-catenin molecular complexes and renal medullary cystic dysplasia in ALK3 transgenic mice. AB - Renal dysplasia, the most frequent cause of childhood renal failure in humans, arises from perturbations in a complex series of morphogenetic events during embryonic renal development. The molecular pathogenesis of renal dysplasia is largely undefined. While investigating the role of a BMP-dependent pathway that inhibits branching morphogenesis in vitro, we generated a novel model of renal dysplasia in a transgenic (Tg) model of ALK3 receptor signaling. We report the renal phenotype, and our discovery of molecular interactions between effectors in the BMP and WNT signaling pathways in dysplastic kidney tissue. Expression of the constitutively active ALK3 receptor ALK3(QD), in two independent transgenic lines caused renal aplasia/severe dysgenesis in 1.5% and 8.4% of hemizygous and homozygous Tg mice, respectively, and renal medullary cystic dysplasia in 49% and 74% of hemizygous and homozygous Tg mice, respectively. The dysplastic phenotype, which included a decreased number of medullary collecting ducts, increased medullary mesenchyme, collecting duct cysts and decreased cortical thickness, was apparent by E18.5. We investigated the pathogenesis of dysplasia in these mice, and demonstrated a 30% decrease in branching morphogenesis at E13.5 before the appearance of histopathogical features of dysplasia, and the formation of beta catenin/SMAD1/SMAD4 molecular complexes in dysplastic renal tissue. Increased transcriptional activity of a beta-catenin reporter gene in ALK3(QD);Tcf-gal mice demonstrated functional cooperativity between the ALK3 and beta-catenin-dependent signaling pathways in kidney tissue. Together with our results in the dysplastic mouse kidney, our findings that phospho-SMAD1 and beta-catenin are overexpressed in human fetal dysplastic renal tissue suggest that dysregulation of these signaling effectors is pathogenic in human renal dysplasia. Our work provides novel insights into the role that crucial developmental signaling pathways may play during the genesis of malformed renal tissue elements. PMID- 12736219 TI - Ectopic expression of kruppel like factor 4 (Klf4) accelerates formation of the epidermal permeability barrier. AB - Dysfunction of the epidermal permeability barrier can result in dehydration, electrolyte imbalance and poor thermoregulation. Immature skin is a portal of entry for infectious agents and potential toxins in topically applied substances. As the skin is one of the last organs to mature in utero, premature infants born before 34 weeks gestation are at great risk for complications. The transcription factor kruppel-like factor 4 (Klf4), has been shown by a targeted ablation, to have an essential function in barrier acquisition. We investigated whether Klf4 expression in utero is sufficient to establish the epidermal barrier. Specifically, we generated lines of mice that express Klf4 from a tetracycline inducible promoter when crossed with transgenic mice expressing the tetracycline transactivator tTA from the epidermal keratin 5 promoter. These mice exhibit acceleration in barrier acquisition as manifest by the exclusion of a dye solution one day earlier in development than controls. Underlying this dye impermeability are morphological changes, including an increased number of stratified layers, expression of terminal differentiation markers and assembly of cornified envelopes. By all criteria, Klf4 ectopic expression accelerates the normal process of terminal differentiation. Premature barrier acquisition in these mice follows the normal pattern rather than the pattern of the transgene promoter, indicating that there are fields of competence in which KLF4 acts. Although other transgenic mice have perturbed barrier acquisition, these mice are the first to accelerate the process of barrier establishment. These studies show that KLF4 regulates barrier acquisition and provides an animal model for studying how to accelerate the process of barrier acquisition for the premature infant. PMID- 12736222 TI - Transcriptional regulation of mitfa accounts for the sox10 requirement in zebrafish melanophore development. AB - The transcription factor Sox10 is required for the specification, migration and survival of all nonectomesenchymal neural crest derivatives including melanophores. sox10(-/-) zebrafish lack expression of the transcription factor mitfa, which itself is required for melanophore development. We demonstrate that the zebrafish mitfa promoter has sox10 binding sites necessary for activity in vitro, consistent with studies using mammalian cell cultures that have shown that Sox10 directly regulates Mitf expression. In addition, we demonstrate that these sites are necessary for promoter activity in vivo. We show that reintroduction of mitfa expression in neural crest cells can rescue melanophore development in sox10(-/-) embryos. This rescue of melanophores in sox10(-/-) embryos is quantitatively indistinguishable from rescue in mitfa(-/-) embryos. These findings show that the essential function of sox10 in melanophore development is limited to transcriptional regulation of mitfa. We propose that the dominant melanophore phenotype in Waardenburg syndrome IV individuals with SOX10 mutations is likely to result from failure to activate MITF in the normal number of melanoblasts. PMID- 12736221 TI - Evidence that Myc activation depletes the epidermal stem cell compartment by modulating adhesive interactions with the local microenvironment. AB - Activation of Myc (c-Myc) causes epidermal cells to exit the stem cell compartment and differentiate into sebocytes and interfollicular epidermis at the expense of the hair lineages. To investigate how Myc exerts these effects we analysed the transcription of more than 10000 genes following Myc activation in the basal layer of mouse epidermis for 1 or 4 days. The major classes of induced genes were involved in synthesis and processing of RNA and proteins, in cell proliferation and in differentiation. More than 40% of the downregulated genes encoded cell adhesion and cytoskeleton proteins. Repression of these genes resulted in profound changes in the adhesive and motile behaviour of keratinocytes. Myc activation inhibited cell motility and wound healing, correlating with decreased expression of a large number of extracellular matrix proteins. Cell adhesion and spreading were also impaired, and this correlated with decreased expression of the alpha6beta4 integrin, decreased formation of hemidesmosomes and decreased assembly of the actomyosin cytoskeleton. We propose that Myc stimulates exit from the stem cell compartment by reducing adhesive interactions with the local microenvironment or niche, and that the failure of hair differentiation reflects an inability of keratinocytes to migrate along the outer root sheath to receive hair inductive stimuli. PMID- 12736223 TI - Current role and future perspectives for ifosfamide in the treatment of malignant non-Hodgkin's lymphoma--results from an expert meeting. PMID- 12736220 TI - Connective tissue growth factor coordinates chondrogenesis and angiogenesis during skeletal development. AB - Coordinated production and remodeling of the extracellular matrix is essential during development. It is of particular importance for skeletogenesis, as the ability of cartilage and bone to provide structural support is determined by the composition and organization of the extracellular matrix. Connective tissue growth factor (CTGF, CCN2) is a secreted protein containing several domains that mediate interactions with growth factors, integrins and extracellular matrix components. A role for CTGF in extracellular matrix production is suggested by its ability to mediate collagen deposition during wound healing. CTGF also induces neovascularization in vitro, suggesting a role in angiogenesis in vivo. To test whether CTGF is required for extracellular matrix remodeling and/or angiogenesis during development, we examined the pattern of Ctgf expression and generated Ctgf-deficient mice. Ctgf is expressed in a variety of tissues in midgestation embryos, with highest levels in vascular tissues and maturing chondrocytes. We confirmed that CTGF is a crucial regulator of cartilage extracellular matrix remodeling by generating Ctgf(-/-) mice. Ctgf deficiency leads to skeletal dysmorphisms as a result of impaired chondrocyte proliferation and extracellular matrix composition within the hypertrophic zone. Decreased expression of specific extracellular matrix components and matrix metalloproteinases suggests that matrix remodeling within the hypertrophic zones in Ctgf mutants is defective. The mutant phenotype also revealed a role for Ctgf in growth plate angiogenesis. Hypertrophic zones of Ctgf mutant growth plates are expanded, and endochondral ossification is impaired. These defects are linked to decreased expression of vascular endothelial growth factor (VEGF) in the hypertrophic zones of Ctgf mutants. These results demonstrate that CTGF is important for cell proliferation and matrix remodeling during chondrogenesis, and is a key regulator coupling extracellular matrix remodeling to angiogenesis at the growth plate. PMID- 12736224 TI - Ifosfamide, carboplatin, etoposide (ICE)-based second-line chemotherapy for the management of relapsed and refractory aggressive non-Hodgkin's lymphoma. AB - Despite advances in the management of aggressive non-Hodgkin's lymphoma, the treatment of relapsed and primary refractory disease remains a major challenge. High-dose chemotherapy or radio-chemotherapy followed by autologous or allogeneic stem cell transplantation (SCT) is a potentially curative treatment approach; however, the applicability of this approach is restricted to patients responding to second-line chemotherapy. Thus, second-line therapy must be both efficacious and without stem cell or organ toxicity that would compromise the ability to proceed to SCT. The ifosfamide, carboplatin and etoposide (ICE) regimen was developed to address these challenges. In a series of prospective clinical trials, 222 patients were treated with the ICE regimen. with an overall response rate of 72%. The mobilization of stem cells with this regimen was excellent,with 86% of patients mobilizing at least 2.0 x 10(6) CD34+ cells/kg. The incidence of treatment-related toxicity precluding SCT after ICE is very low. Herein, we report the clinical results of this treatment program for 222 patients with 5 year median follow-up for surviving patients. Rituximab was subsequently added to the ICE regimen for patients with diffuse large B-cell lymphoma (DLBCL) to improve upon these favorable results. This resulted in an increased complete remission rate. Additional follow-up is necessary to determine if this improvement in the complete remission rate will confer an increase in the overall survival following SCT. PMID- 12736225 TI - Outpatient-based ifosfamide, carboplatin and etoposide (ICE) chemotherapy in transplant-eligible patients with non-Hodgkin's lymphoma and Hodgkin's disease. AB - We have treated 38 transplant-eligible patients with relapsed/refractory non Hodgkin's lymphoma and Hodgkin's disease using an outpatient-based regimen of ifosfamide, carboplatin and etoposide (ICE) for both salvage and peripheral blood stem cell mobilisation. Patients included relapsed or refractory diffuse large B cell lymphoma (n = 17), follicular lymphoma (n = II), T-cell lymphoma (n = 2), mantle cell lymphoma (n = 2) and Hodgkin's disease (n = 6). Seven patients with diffuse large B-cell lymphoma and three patients with follicular lymphoma (26%) were considered chemorefractory. Cycles of ICE therapy were administered every 21 days as an outpatient and consisted of ifosfamide 5000 mg/m2 intravenously (i.v.) fractionated into three equally divided doses over 3 days, carboplatin [mg dose = 5 x area under the curve (AUC)] i.v. on day 1 and etoposide 100 mg/m2- i.v. daily for 3 days. Subsequently. granulocyte colony-stimulating factor (G-CSF)5 microg/kg subcutaneously (s.c.) was administered daily from day +5. Of the I I follicular lymphoma patients, 10 also received rituximab with ICE therapy. Median age of patients was 52 years (range 30-65). Patients received a mean of 2.6 cycles (range 1-4) of ICE. There were no toxic deaths and no significant non haematological toxicities secondary to ICE therapy. Grade IV thrombocytopenia and grade IV neutropenia with at least one cycle of ICE were seen in 47% and 53% of patients, respectively. Median time to peripheral blood stem cell (PBSC) harvest was 14 days (range 10-20). while the median CD34+ cell yield was 5.2 x 10(6) cells/kg(range 2.3 x 10(6)-27.2 x 10(6)). Only one of the ICE-responders failed to mobilise PBSCs. The overall response rate to ICE was 87%. comprising 14 patients (37%) who achieved a complete response (CR) and 19 (50%) who achieved a partial response (PR). A total of 30 patients have undergone autologous stem cell transplantation(SCT) while two follicular lymphoma patients have received a non myeloablative allogeneic SCT. Follow-up is short: however, the Kaplan-Meier estimate of the proportion of patients alive and event-free at a median follow-up of 11 months is 80% and 59%, respectively. Event-free survival for patients who achieved a CR after ICE and transplantation is 88% versus 45% for those who achieved a PR. These data confirm the efficacy and tolerability of fractionated ICE chemotherapy as both a salvage and mobilisation regimen that can be readily delivered in an outpatient setting. PMID- 12736227 TI - A phase I-II study of rituximab, ifosfamide, mitoxantrone and etoposide (R-IME) for B cell non-Hodgkin's lymphoma prior to and after high-dose chemotherapy and autologous stem cell transplantation (HDC-ASCT). AB - This phase I-II study describes the safety of rituximab, ifosfamide, mitoxantrone and etoposide (R-IME) as an induction regimen prior to high-dose chemotherapy and autologous stem cell transplantation (HDC-ASCT), and rituximab given post-HDC ASCT for B cell non-Hodgkins's lymphoma. This study also measured the effect on disease burden and stem cell contamination. Patients with relapsed, refractory or poor risk B cell lymphomas were eligible. Patients were treated with two cycles of R-IME; all non-progressing patients under-went a third cycle and peripheral blood stem cell (PBSC) collection. Patients underwent HDC-ASCT and those patients in remission after HDC-ASCT were treated with four additional doses of rituximab. Tumor cell contamination was measured at baseline and in the PBSC. Serial immunoglobulin levels were measured. Patients were followed for time to treatment failure (TTF) and overall survival (OS). Thirty-two patients were enrolled. Thirty patients had at least stable disease after two cycles of R-IME. Twenty nine underwent stem cell collection. The response rate to R-IME induction was 77% (20/26) with 35% (9/26) complete response(CR). Stem cell mobilization was successful in 93% (27/29) of patients. The response rate to R-IME induction and HDC-ASCT was 95% with a confirmed CR of 68%. Median follow-up was 28 months; the median TTFand OS have not been reached. There was a significant decline in stem cell tumor cell contamination and a significant decline in IgG without an increase in infections. Forty-three per cent of patients had transient neutropenia after post-transplant rituximab. R-IME is an effective cytoreductive and mobilization regimen. There appears to be a reduction in the number of lymphoma cells in the stem cell product and the toxicity is manageable. PMID- 12736226 TI - Outpatient regimen rituximab plus ifosfamide, carboplatin and etoposide (R-ICE) for relapsed non-Hodgkin's lymphoma. PMID- 12736229 TI - Hodgkin's lymphoma: what is next in salvage treatment? PMID- 12736228 TI - Impact of chemotherapy regimen and hematopoietic growth factor on mobilization and collection of peripheral blood stem cells in cancer patients. AB - Various chemotherapy regimens, combined with recombinant human granulocyte colony stimulating factor(rhG-CSF) or recombinant granulocyte-macrophage CSF (rhGM-CSF) are used in cancer patients to mobilize and collect peripheral blood stem cells (PBSC). In this retrospective study, we evaluated and compared the efficacy of such regimens in 262 patients with different types of malignant diseases. The following chemotherapy regimens were applied: ifosfamide-etoposide-cisplatin or bleomycin (n = 96; mainly patients with testicular cancer); ifosfamide-etoposide plus or minus cytosine arabinoside (Ara-C) or vincristine (VCR)(n = 52; mainly patients with lymphoma); cyclophosphamide-anthracycline (n = 53; mainly patients with breast cancer); intermediate to high dose (ID-HD) cyclophosphamide (n = 37; mainly patients with breast or ovarian cancer. or multiple myeloma; and others (n = 24). rhG-CSF or rhGM-CSF, each at an average daily dose of 5 microg/kg body weight, were used in 166 and 96 patients, respectively. The study evaluated and compared the efficacy of these two cytokines. In patients receiving rhG-CSF, CD34+ cells could be collected earlier (median: day 14 versus day 16) and there was a significantly higher white blood cell count (WBC)(median 11,350 versus 5550/microl) and CD34+ cell count (median 88 versus 43/microl) at the start of apheresis, and a significantly higher CD34+ cell yield (median 7.4 x 10(6) versus 4.6 x 10(6)/kg) than in patients who receivedrhGM-CSF. Among the various chemotherapeutic regimens used, each combined with rhG-CSF, ifosfamide-etoposide plus or minus Ara-C or VCR mobilized a significantly higher number of CD34+ cells (median 119/microl) and produced a significantly higher harvest of these cells (median 13 x 10(6)/kg) than cyclophosphamide-anthracycline (median 87/microl and 7 x 10(6)/kg, respectively) or ID-HD cyclophosphamide (median 59/microl and 5 x I 0(6)/kg, respectively). Ifosfamide-etoposide plus or minus Ara-C or VCR was also superior to ifosfamide-etoposide-cisplatin or bleomycin (median 78/microl and 9 x 10(6)/kg, respectively), but at borderline significance. The outcome of PBSC mobilization and collection appeared to be negatively influenced by the number of relapses before the current salvage treatment. These data indicate that mobilization and collection of PBSCstrongly depend on the type of hematopoietic growth factor and chemotherapeutic regimen used. The data further show rhG-CSF is a more effective growth factor than rhGM-CSF and ifosfamide-etoposide-based regimens, particularly ifosfamide-etoposide plus or minus Ara-C or VCR, are highly effective regimens in mobilizing and collecting CD34+ cells. PMID- 12736230 TI - Ifosfamide salvage treatment before autologous stem cell transplantation in patients with refractory and relapsed Hodgkin's lymphoma: a GELA experience. PMID- 12736231 TI - Ifosfamide, epirubicin and etoposide (IEV) in non-Hodgkin's lymphoma and Hodgkin's disease: the Italian experience. PMID- 12736232 TI - Strategic approach to the management of Hodgkin's disease incorporating salvage therapy with high-dose ifosfamide, etoposide and epirubicin: a Northern Region Lymphoma Group study (UK). AB - The Northern Region Lymphoma Group is a population-based group covering 3.1 million people in Northern England. From 1991 total data collection for all Hodgkin's disease patients for this population has been in place and it has been possible to demonstrate that the overall survival for Hodgkin's disease for younger patients within this population has moved from 80% pre- 1988 to 87% post- 1988. This improvement has been brought about by the introduction of clinical trials for advanced stage disease and effective salvage regimens. This report describes the outcome of 51 patients treated with the ifosfamide, etoposide and epirubicin (IVE)schedule and includes 28 males and 23 females with a median age of 34 years. Overall 43 of 51 patients responded to treatment (84%) with 31 achieving a complete response, four a good partial response and eight a partial response. Thirty-one proceeded to autologous stem-cell transplantation. In total, with a median follow-up of 24 months (range 6-51), 26 patients remain alive and in continuous remission. Haematological toxicity,in particular neutropenia WHO grade 4, was observed in all cases but improved over the three courses of treatment. Non-haematological toxicity was not a major problem, with no significant cardiac, hepatic, renal or neurotoxicity. We conclude that the high dose ifosfamide-containing regimens should be prospectively evaluated in the various types of non-responsive and relapsing Hodgkin's disease. PMID- 12736233 TI - Saccade-related activity in the primate superior colliculus depends on the presence of local landmarks at the saccade endpoint. AB - Saccade-related discharge in the superior colliculus is greater for saccades made to a spot of light than for saccades in complete darkness. However, it is unclear whether this enhancement is due to the discontinuity of the spot or due to its being a new object of fixation. In these experiments, we examined the saccade related activity of intermediate-layer neurons in the primate superior colliculus during delayed saccades to the center or corner of a large, bright square, as well as for visual and memory-guided movements to small spots in isolation. The saccade-related discharge for movements made to a local visual landmark present at the time of the saccade, be it a corner of a square or a small spot, was higher than that for saccades made to the center of a square that contained no local visual landmarks within. Moreover, discharge for movements to the center of a square were very similar to that for saccades to blank, dark space. Saccade velocity was similarly dependent on the presence of such a landmark, though less dramatically. The endpoints of saccades directed toward a square's corner were slightly displaced toward the center of the square. Across all neurons, discharge and velocity for saccades to the center of a square increased as the square size was decreased, but were never greater than those for saccades to a small spot of light. These results suggest that both saccade-related discharge in the superior colliculus and saccade metrics are enhanced for movements directed to parts of the visual scene with high contrast, while shifting fixation to a new object is not itself sufficient to elevate discharge and metrics above those of saccades to blank space. PMID- 12736234 TI - Macaque frontal eye field input to saccade-related neurons in the superior colliculus. AB - Extracellular recordings were made simultaneously in the frontal eye field and superior colliculus in awake, behaving rhesus monkeys. Frontal eye field microstimulation was used to orthodromically activate the superior colliculus both to locate the depth of the strongest frontal eye field input to the superior colliculus and to identify superior colliculus neurons receiving direct frontal eye field input. The activity of orthodromically driven colliculus neurons was characterized during visuomotor tasks. The purpose of this study was to identify the types of superior colliculus neurons that receive excitatory frontal eye field input. We found that microstimulation of the frontal eye field did not activate the superficial layers of the superior colliculus but did activate the deeper layers. This pattern of activation coincided with the prevalence of visual versus saccade-related activity in the superficial and deep layers. A total of 83 orthodromically driven superior colliculus neurons were identified. Of these neurons, 93% (n = 77) exhibited a burst of activity associated with the onset of the saccade, and 25% (n = 21) exhibited prelude/build-up activity prior to the onset of a saccade. In addition, it was common to see some activity synchronized with the onset of a visual target (30%, n = 25). In single neurons, these activity profiles could be observed alone or in combination. Superior colliculus neurons that were exclusively visual, however, were not excited by frontal eye field stimulation. We compared the activity of superior colliculus neurons that received frontal eye field input to descriptions of saccade-related neurons made in earlier reports and found that the distribution of neuron types in the orthodromically driven population was similar to the distribution within the overall population. This suggests that the frontal eye field does not selectively influence a specific class of collicular neurons, but, instead has a direct influence on all preparatory, and saccade-related activity within the deep layers of the superior colliculus. PMID- 12736235 TI - From rule to response: neuronal processes in the premotor and prefrontal cortex. AB - The ability to use abstract rules or principles allows behavior to generalize from specific circumstances (e.g., rules learned in a specific restaurant can subsequently be applied to any dining experience). Neurons in the prefrontal cortex (PFC) encode such rules. However, to guide behavior, rules must be linked to motor responses. We investigated the neuronal mechanisms underlying this process by recording from the PFC and the premotor cortex (PMC) of monkeys trained to use two abstract rules: "same" or "different." The monkeys had to either hold or release a lever, depending on whether two successively presented pictures were the same or different, and depending on which rule was in effect. The abstract rules were represented in both regions, although they were more prevalent and were encoded earlier and more strongly in the PMC. There was a perceptual bias in the PFC, relative to the PMC, with more PFC neurons encoding the presented pictures. In contrast, neurons encoding the behavioral response were more prevalent in the PMC, and the selectivity was stronger and appeared earlier in the PMC than in the PFC. PMID- 12736236 TI - Salt-evoked lingual surface potential in humans. AB - Salt sensing in animals involves the epithelial sodium channel (ENaC). If ENaC were involved in human salt sensing, then the lingual surface potential (LSP) would hyperpolarize when exposed to sodium. We developed a chamber to measure the LSP while different solutions superfused the surface of the tongue and a technique to adjust for the junction potentials induced by varying salt concentrations. Changing the superfusion solution from rinse solution (30 mM KCl) to 300 mM NaCl (+30 mM KCl) caused the LSP to hyperpolarize by 10.1 +/- 0.7 mV (n = 13, P < 0.001). With repeated challenge the LSP response was reproducible. Increasing the Na concentration from 100 to 600 mM increased hyperpolarization by 35 +/- 4.8% (n = 9, P < 0.001). To examine whether amiloride affects the LSP, 0.1 mM amiloride was added to 300 mM NaCl; it reduced the hyperpolarization by 18.5 +/- 4.3% (P < 0.005, n = 11). However, the amiloride effect was not uniform: in six volunteers, amiloride inhibited the LSP by as much as 42%, while in five subjects, amiloride inhibited <5% of the LSP. In an amiloride sensitive volunteer, amiloride exerted 50% of its effect at 1 microM. In conclusion, we have demonstrated that the LSP can be measured in humans, that Na hyperpolarizes the LSP, that increasing the Na concentration increases LSP hyperpolarization, and that amiloride inhibits the Na evoked LSP in some humans. While ENaC is involved in sensing salt, its role appears to vary among individuals. PMID- 12736237 TI - Nondominant arm advantages in load compensation during rapid elbow joint movements. AB - This study was designed to examine interlimb asymmetries in responding to unpredictable changes in inertial loads, which have implications for our understanding of the neural mechanisms underlying handedness. Subjects made repetitive single joint speed constrained 20 degrees elbow flexion movements, while the arm was supported on a horizontal, frictionless, air-jet system. On random trials, a 2-kg mass was attached to the arm splint prior to the "go" signal. Subjects were not given explicit information about the mass prior to movement nor were they able to view their limb or the mass. Accordingly, muscle activity, recorded prior to peak tangential finger acceleration, was the same for loaded and baseline trials. After this point, substantial changes in muscle activity occurred. In both limbs, the load compensation response was associated with a reduction in extensor muscle activity, resulting in a prolonged flexion phase of motion. For the nondominant arm, this resulted in effective load compensation, such that no differences in final position accuracy occurred between loaded and baseline trials. However, the dominant arm response also included a considerable increase in flexor muscle activity. This substantially prolonged the flexor acceleration phase of motion, relative to that of the nondominant arm. As a result, the dominant arm overcompensated the effects of the load, producing a large and systematic overshoot of final position. These results indicate more effective load compensation responses for the nondominant arm; supporting a specialized role of the nondominant arm/hemisphere system in sensory feedback mediated error correction mechanisms. The results also suggest that specialization of the dominant arm system for controlling limb and task dynamics is specifically related to feedforward control mechanisms. PMID- 12736238 TI - Differential contribution of reticulospinal cells to the control of locomotion induced by the mesencephalic locomotor region. AB - In lampreys as in other vertebrates, the reticulospinal (RS) system relays inputs from the mesencephalic locomotor region (MLR) to the spinal locomotor networks. Semi-intact preparations of larval sea lamprey were used to determine the relative contribution of the middle (MRRN) and the posterior (PRRN) rhombencephalic reticular nuclei to swimming controlled by the MLR. Intracellular recordings were performed to examine the inputs from the MLR to RS neurons. Stimulation of the MLR elicited monosynaptic excitatory responses of a higher magnitude in the MRRN than in the PRRN. This differential effect was not attributed to intrinsic properties of RS neurons. Paired recordings showed that at threshold intensity for swimming, spiking activity was primarily elicited in RS cells of the MRRN. Interestingly, cells of the PRRN began to discharge at higher stimulation intensities only when MRRN cells had reached their maximal discharge rate. Glutamate antagonists were ejected in either nucleus to reduce their activity. Ejections over the MRRN increased the stimulation threshold for evoking locomotion and resulted in a marked decrease in the swimming frequency and the strength of the muscle contractions. Ejections over the PRRN decreased the frequency of swimming. This study provides support for the concept that RS cells show a specific recruitment pattern during MLR-induced locomotion. RS cells in the MRRN are primarily involved in initiation and maintenance of low-intensity swimming. At higher frequency locomotor rhythm, RS cells in both the MRRN and the PRRN are recruited. PMID- 12736240 TI - Visual tracking neurons in primate area MST are activated by smooth-pursuit eye movements of an "imaginary" target. AB - Because smooth-pursuit eye movements (SPEM) can be executed only in the presence of a moving target, it has been difficult to attribute the neuronal activity observed during the execution of these eye movements to either sensory processing or to motor preparation or execution. Previously, we showed that rhesus monkeys can be trained to perform SPEM directed toward an "imaginary" target defined by visual cues confined to the periphery of the visual field. The pursuit of an "imaginary" target provides the opportunity to elicit SPEM without stimulating visual receptive fields confined to the center of the visual field. Here, we report that a subset of neurons [85 "imaginary" visual tracking (iVT)-neurons] in area MST of 3 rhesus monkeys were identically activated during pursuit of a conventional, foveal dot target and the "imaginary" target. Because iVT-neurons did not respond to the presentation of a moving "imaginary" target during fixation of a stationary dot, we are able to exclude that responses to pursuit of the "imaginary" target were artifacts of stimulation of the visual field periphery. Neurons recorded from the representation of the central parts of the visual field in neighboring area MT, usually vigorously discharging during pursuit of foveal targets, in no case responded to pursuit of the "imaginary" target. This dissociation between MT and MST neurons supports the view that pursuit responses of MT neurons are the result of target image motion, whereas those of iVT-neurons in area MST reflect an eye movement-related signal that is nonretinal in origin. iVT-neurons fell into two groups, depending on the properties of the eye movement-related signal. Whereas most of them (71%) encoded eye velocity, a minority showed responses determined by eye position, irrespective of whether eye position was changed by smooth pursuit or by saccades. Only the former group exhibited responses that led the eye movement, which is a prerequisite for a causal role in the generation of SPEM. PMID- 12736239 TI - Robustness of the tuning of fly visual interneurons to rotatory optic flow. AB - The sophisticated receptive field organization of motion-sensitive tangential cells in the visual system of the blowfly Calliphora vicina matches the structure of particular optic flow fields. Hypotheses on the tuning of particular tangential cells to rotatory self-motion are based on local motion measurements. So far, tangential cells have never been tested with global optic flow stimuli. Therefore we measured the responses of an identifiable neuron, the V1 tangential cell, to wide-field motion stimuli mimicking optic flow fields similar to those the fly encounters during particular self-motions. The stimuli were generated by a "planetarium-projector," casting a pattern of moving light dots on a large spherical projection screen. We determined the tuning curves of the V1-cell to optic flow fields as induced by the animal during 1) rotation about horizontally aligned body axes, 2) upward/downward translation, and 3) a combination of both components. We found that the V1-cell does not respond as specifically to self rotations, as had been concluded from its receptive field organization. The neuron responds strongly to upward translation and its tuning to rotations is much coarser than expected. The discrepancies between the responses to global optic flow and the predictions based on the receptive field organization are likely due to nonlinear integration properties of tangential neurons. Response parameters like orientation, shape, and width of the tuning curve are largely unaffected by changes in rotation velocity or a superposition of rotational and translational optic flow. PMID- 12736241 TI - Dendrodendritic inhibition and simulated odor responses in a detailed olfactory bulb network model. AB - In the olfactory bulb, both the spatial distribution and the temporal structure of neuronal activity appear to be important for processing odor information, but it is currently impossible to measure both of these simultaneously with high resolution and in all layers of the bulb. We have developed a biologically realistic model of the mammalian olfactory bulb, incorporating the mitral and granule cells and the dendrodendritic synapses between them, which allows us to observe the network behavior in detail. The cell models were based on previously published work. The attributes of the synapses were obtained from the literature. The pattern of synaptic connections was based on the limited experimental data in the literature on the statistics of connections between neurons in the bulb. The results of simulation experiments with electrical stimulation agree closely in most details with published experimental data. This gives confidence that the model is capturing features of network interactions in the real olfactory bulb. The model predicts that the time course of dendrodendritic inhibition is dependent on the network connectivity as well as on the intrinsic parameters of the synapses. In response to simulated odor stimulation, strongly activated mitral cells tend to suppress neighboring cells, the mitral cells readily synchronize their firing, and increasing the stimulus intensity increases the degree of synchronization. Preliminary experiments suggest that slow temporal changes in the degree of synchronization are more useful in distinguishing between very similar odorants than is the spatial distribution of mean firing rate. PMID- 12736243 TI - Intrinsic properties and reflex compensation in reinnervated triceps surae muscles of the cat: effect of movement history. AB - Effects of prior motion on ramp stretch responses of reflexive and areflexive muscles were measured in decerebrate cats. Soleus and gastrocnemius muscles were rendered areflexive by reinnervation a minimum of 9 mo before the terminal experiments. The introduction of a shortening phase prior to the ramp stretch increased the normalized initial stiffness of muscles and decreased the tendency to yield of the reinnervated muscles as compared with the case in which muscles contracted isometrically prior to stretch. Yielding was compensated by reflex action for all amplitudes of prior shortening in soleus and gastrocnemius muscles. The comparison of responses of untreated and reinnervated muscles indicated that the contribution of reflex action progressively declined with the amplitude of prior shortening as the extent of yielding diminished. In soleus muscle, during a variable delay period of isometric contraction interposed between shortening and lengthening force generation, initial stiffness and yielding returned to levels seen with isometric contractile history. However, these attributes recovered at different rates, suggesting that distinct processes are responsible for initial stiffness and yielding. Yielding was compensated for by reflex action regardless of the length of the interposed delay or of the amplitude of the prior shortening. These and previous findings indicate that the stretch reflex regulates muscular stiffness for a wide range of conditions. This regulation apparently arises from complementary mechanical properties of intrafusal and extrafusal muscle. PMID- 12736242 TI - Intrinsic properties and reflex compensation in reinnervated triceps surae muscles of the cat: effect of activation level. AB - The manner in which activation levels influence intrinsic muscular properties and contributions of the stretch reflex were studied in homogeneous soleus (SOL) and heterogeneous gastrocnemius (G) muscles in the decerebrate cat. Intrinsic mechanical properties were represented by the initial stiffness of the muscle, measured prior to reflex action, and by the tendency of the muscle to yield during stretch in the absence of the stretch reflex. Stiffness regulation by the stretch reflex was evaluated by measuring the extent to which reflex action reduces yielding and the extent to which stiffness depends on background force. Intrinsic mechanical properties were measured in muscles deprived of effective autogenic reflexes using the method of muscular reinnervation. Reinnervated muscles were recruited to force levels comparable to those achieved during natural locomotion. As force declined during crossed-extension reflexes in reinnervated and intact muscles, initial stiffness declined according to similar convex trajectories. The data did not support the hypothesis that, for a given force level, initial stiffness is greatest in populations of predominantly type I motor units. Incremental stiffness (Deltaf/Deltal) of both G and SOL increased in the presence of the stretch reflex. Yielding of SOL (ratio of incremental to initial stiffness) substantially decreased in the presence of the stretch reflex over the full range of forces. In reflexive G, yielding significantly decreased for low to intermediate forces, whereas at higher forces, yielding was similar irrespective of the presence or absence of the stretch reflex. The stretch reflex regulates stiffness in both homogeneous and heterogeneous muscles. PMID- 12736245 TI - Binding of the chemotaxis response regulator CheY to the isolated, intact switch complex of the bacterial flagellar motor: lack of cooperativity. AB - In bacteria, the chemotactic signal is greatly amplified between the chemotaxis receptors and the flagellar motor. In Escherichia coli, part of this amplification occurs at the flagellar switch. However, it is not known whether the amplification results from cooperativity of CheY binding to the switch or from a post-binding step. To address this question, we purified the intact switch complex (constituting the switch proteins FliG, FliM, and FliN and the scaffolding protein FliF) in quantities sufficient for biochemical work and used it to investigate whether the binding of CheY to the switch complex is cooperative. As a negative control, we used complexes of switchless basal bodies, formed from the proteins FliF and FliG and similarly isolated. Using double labeling centrifugation assays for binding, we found that CheY binds to the isolated, intact switch complex in a phosphorylation-dependent manner. We observed no significant phosphorylation-dependent binding to the negative control of the switchless basal body. The dissociation constant for the binding between the switch complex and phosphorylated CheY (CheY approximately P) was 4.0 +/- 1.1 microm, well in line with the published range of CheY approximately P concentrations to which the flagellar motor is responsive. Furthermore, the binding was not cooperative (Hill coefficient approximately 1). This lack of CheY approximately P-switch complex binding cooperativity, taken together with earlier in vivo studies suggesting that the dependence of the rotational state of the motor on the fraction of occupied sites at the switch is sigmoidal and very steep (Bren, A., and Eisenbach, M. (2001) J. Mol. Biol. 312, 699-709), indicates that the chemotactic signal is amplified within the switch, subsequent to the CheY approximately P binding. PMID- 12736244 TI - The voltage-dependent anion channel is a receptor for plasminogen kringle 5 on human endothelial cells. AB - Human plasminogen contains structural domains that are termed kringles. Proteolytic cleavage of plasminogen yields kringles 1-3 or 4 and kringle 5 (K5), which regulate endothelial cell proliferation. The receptor for kringles 1-3 or 4 has been identified as cell surface-associated ATP synthase; however, the receptor for K5 is not known. Sequence homology exists between the plasminogen activator streptokinase and the human voltage-dependent anion channel (VDAC); however, a functional relationship between these proteins has not been reported. A streptokinase binding site for K5 is located between residues Tyr252-Lys283, which is homologous to the primary sequence of VDAC residues Tyr224-Lys255. Antibodies against these sequences react with VDAC and detect this protein on the plasma membrane of human endothelial cells. K5 binds with high affinity (Kd of 28 nm) to endothelial cells, and binding is inhibited by these antibodies. Purified VDAC binds to K5 but only when reconstituted into liposomes. K5 also interferes with mechanisms controlling the regulation of intracellular Ca2+ via its interaction with VDAC. K5 binding to endothelial cells also induces a decrease in intracellular pH and hyperpolarization of the mitochondrial membrane. These studies suggest that VDAC is a receptor for K5. PMID- 12736246 TI - Engineering of a staphylokinase-based fibrinolytic agent with antithrombotic activity and targeting capability toward thrombin-rich fibrin and plasma clots. AB - Current clinically approved thrombolytic agents have significant drawbacks including reocclusion and bleeding complications. To address these problems, a staphylokinase-based thrombolytic agent equipped with antithrombotic activity from hirudin was engineered. Because the N termini for both staphylokinase and hirudin are required for their activities, a Y-shaped molecule is generated using engineered coiled-coil sequences as the heterodimerization domain. This agent, designated HE-SAKK, was produced and assembled from Bacillus subtilis via secretion using an optimized co-cultivation approach. After a simple in vitro treatment to reshuffle the disulfide bonds of hirudin, both staphylokinase and hirudin in HE-SAKK showed biological activities comparable with their parent molecules. This agent was capable of targeting thrombin-rich fibrin clots and inhibiting clot-bound thrombin activity. The time required for lysing 50% of fibrin clot in the absence or presence of fibrinogen was shortened 21 and 30%, respectively, with HE-SAKK in comparison with staphylokinase. In plasma clot studies, the HE-SAKK concentration required to achieve a comparable 50% clot lysis time was at least 12 times less than that of staphylokinase. Therefore, HE SAKK is a promising thrombolytic agent with the capability to target thrombin rich fibrin clots and to minimize clot reformation during fibrinolysis. PMID- 12736247 TI - Model glycosulfopeptides from P-selectin glycoprotein ligand-1 require tyrosine sulfation and a core 2-branched O-glycan to bind to L-selectin. AB - L-selectin expressed on leukocytes is involved in lymphocyte homing to secondary lymphoid organs and leukocyte recruitment into inflamed tissue. L-selectin binds to the sulfated sialyl Lewis x (6-sulfo-sLex) epitope present on O-glycans of various glycoproteins in high endothelial venules. In addition, L-selectin interacts with the dimeric mucin P-selectin glycoprotein ligand-1 (PSGL-1) expressed on leukocytes. PSGL-1 lacks 6-sulfo-sLex but contains sulfated tyrosine residues (Tyr-SO3)at positions 46, 48, and 51 and sLex in a core 2-based O-glycan (C2-O-sLex) on Thr at position 57. The role of tyrosine sulfation and core 2 O glycans in binding of PSGL-1 to L-selectin is not well defined. Here, we show that L-selectin binds to a glycosulfopeptide (GSP-6) modeled after the extreme N terminus of human PSGL-1, containing three Tyr-SO3 and a nearby Thr modified with C2-O-sLex. Leukocytes roll on immobilized GSP-6 in an L-selectin-dependent manner, and rolling is dependent on Tyr-SO3 and C2-O-sLex on GSP-6. The dissociation constant for binding of L-selectin to GSP-6, as measured by equilibrium gel filtration, is approximately 5 microm. Binding is dependent on Tyr-SO3 residues as well as the sialic acid and fucose residues of C2-O-sLex. Binding to an isomeric glycosulfopeptide containing three Tyr-SO3 residues and a core 1-based O-glycan expressing sLex was reduced by approximately 90%. All three Tyr-SO3 residues of GSP-6 are required for high affinity binding to L-selectin. Low affinity binding to mono- and disulfated GSPs is largely independent of the position of the Tyr-SO3 residues, except for some binding preference for an isomer sulfated on both Tyr-48 and -51. These results demonstrate that L-selectin binds with high affinity to the N-terminal region of PSGL-1 through cooperative interactions with three sulfated tyrosine residues and an appropriately positioned C2-O-sLex O-glycan. PMID- 12736248 TI - Dexamethasone-induced gene 2 (dig2) is a novel pro-survival stress gene induced rapidly by diverse apoptotic signals. AB - Glucocorticoid hormones induce apoptosis in lymphoid cells. This process requires de novo RNA/protein synthesis. Here we report the identification and cloning of a novel dexamethasone-induced gene designated dig2. Using Affymetrix oligonucleotide microarray analysis of approximately 10,000 genes and expressed sequence tags, we found that the expression of dig2 mRNA is significantly induced not only in the murine T cell lymphoma lines S49.A2 and WEHI7.2 but also in normal mouse thymocytes following dexamethasone treatment. This result was confirmed by Northern blot analysis. The induction of dig2 mRNA by dexamethasone appears to be mediated through the glucocorticoid receptor as it is blocked in the presence of RU486, a glucocorticoid receptor antagonist. Furthermore, we demonstrated that dig2 is a novel stress response gene, as its mRNA is induced in response to a variety of cellular stressors including thapsigargin, tunicamycin, and heat shock. In addition, the levels of dig2 mRNA were up-regulated after treatment with the apoptosis-inducing chemotherapeutic drug etoposide. Though the function of dig2 is unknown, dig2 appears to have a pro-survival function, as overexpression of dig2 reduces the sensitivity of WEHI7.2 cells to dexamethasone induced apoptosis. PMID- 12736249 TI - Ouabain is a potent promoter of growth and activator of ERK1/2 in ouabain resistant rat renal epithelial cells. AB - Endogenous cardiotonic steroids (ECS) are putative ligands of the inhibitory binding site of the membrane sodium pump (Na+, K+-ATPase). There is growing evidence that cardiotonic steroids may promote the growth of cardiac and vascular myocytes, including evidence indicating growth stimulation at concentrations in the same range as circulating ECS concentrations. We investigated four parameters to determine whether ouabain, a proposed ECS, promotes growth of immortalized rat proximal tubule epithelial cells: cell count by hemocytometer; metabolic activity as reflected in the mitochondrial conversion of the tetrazolium salt, 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, to its formazan product (MA); DNA synthesis reflected as bromodeoxyuridine incorporation (DNA); and mitosis reflected as histone phosphorylation state detected using anti phosphohistone 3 antibody (HP). Maximum stimulatory responses were observed at 1 nm ouabain (MA, 20.3% increase, p < 0.01; DNA, 28.4% increase, p < 0.001; HP, maximum response at 0.5 h, 50% increase, p < 0.001). We observed that growth stimulation was associated with stimulation of ERK1/2 phosphorylation (ERK-P), and both growth and ERK-P could be blocked by the MEK inhibitor (U0126, 100 nm). Western blot analysis revealed that the only alpha isoform of Na+, K+-ATPase that could be detected in these cultures was the highly ouabain-resistant alpha1 isoform. Measurement of ouabain inhibition of ion transport in these cultures using 86Rb+ uptake revealed the predominance of the expected ouabain-resistant isoform (IC50 = 24 microm) and an additional minor ( approximately 15%) ouabain sensitive inhibition with IC50 approximately 30 pm. Similar bimodal transport inhibition curves were obtained in freshly dissected rat proximal tubules. These results indicate that renal epithelial cells may be a sensitive target of the ERK1/2-activating and growth-promoting effects of ouabain even in the presence of ouabain-resistant Na+, K+-ATPase. PMID- 12736250 TI - Presenilin-1, nicastrin, amyloid precursor protein, and gamma-secretase activity are co-localized in the lysosomal membrane. AB - Alzheimer's disease (AD) is caused by the cerebral deposition of beta-amyloid (Abeta), a 38-43-amino acid peptide derived by proteolytic cleavage of the amyloid precursor protein (APP). Initial studies indicated that final cleavage of APP by the gamma-secretase (a complex containing presenilin and nicastrin) to produce Abeta occurred in the endosomal/lysosomal system. However, other studies showing a predominant endoplasmic reticulum localization of the gamma-secretase proteins and a neutral pH optimum of in vitro gamma-secretase assays have challenged this conclusion. We have recently identified nicastrin as a major lysosomal membrane protein. In the present work, we use Western blotting and immunogold electron microscopy to demonstrate that significant amounts of mature nicastrin, presenilin-1, and APP are co-localized with lysosomal associated membrane protein-1 (cAMP-1) in the outer membranes of lysosomes. Furthermore, we demonstrate that these membranes contain an acidic gamma-secretase activity, which is immunoprecipitable with an antibody to nicastrin. These experiments establish APP, nicastrin, and presenilin-1 as resident lysosomal membrane proteins and indicate that gamma-secretase is a lysosomal protease. These data reassert the importance of the lysosomal/endosomal system in the generation of Abeta and suggest a role for lysosomes in the pathophysiology of AD. PMID- 12736251 TI - Identification of a novel integrin alphaMbeta2 binding site in CCN1 (CYR61), a matricellular protein expressed in healing wounds and atherosclerotic lesions. AB - CCN1 (cysteine-rich 61) and CCN2 (connective tissue growth factor) are growth factor-inducible immediate-early gene products found in atherosclerotic lesions, restenosed blood vessels, and healing cutaneous wounds. Both CCN proteins have been shown to support cell adhesion and induce cell migration through interaction with integrin receptors. Recently, we have identified integrin alphaMbeta2 as the major adhesion receptor mediating monocyte adhesion to CCN1 and CCN2 and have shown that the alphaMI domain binds specifically to both proteins. In the present study, we demonstrated that activated monocytes adhered to a synthetic peptide (CCN1-H2, SSVKKYRPKYCGS) derived from a conserved region within the CCN1 C terminal domain, and this process was blocked by the anti-alphaM monoclonal antibody 2LPM19c. Consistently, a glutathione S-transferase (GST) fusion protein containing the alphaMI domain (GST-alphaMI) bound to immobilized CCN1-H2 as well as to the corresponding H2 sequence in CCN2 (CCN2-H2, TSVKTYRAKFCGV). By contrast, a scrambled CCN1-H2 peptide and an 18-residue peptide derived from an adjacent sequence of CCN1-H2 failed to support monocyte adhesion or alphaMI domain binding. To confirm that the CCN1-H2 sequence within the CCN1 protein mediates alphaMbeta2 interaction, we developed an anti-peptide antibody against CCN1-H2 and showed that it specifically blocked GST-alphaMI binding to intact CCN1. Collectively, these results identify the H2 sequence in CCN1 and CCN2 as a novel integrin alphaMbeta2 binding motif that bears no apparent homology to any alphaMbeta2 binding sequence reported to date. PMID- 12736252 TI - IKKi/IKKepsilon plays a key role in integrating signals induced by pro inflammatory stimuli. AB - We report that the product of the inducible gene encoding the kinase known as IKKi/IKKepsilon (IKKi) is required for expression of a group of genes up regulated by pro-inflammatory stimuli such as bacterial endotoxin (lipopolysaccharide (LPS)). Here, using murine embryonic fibroblasts obtained from mice bearing deletions in IKK2, p65, and IKKi genes, we provide evidence to support a link between signaling through the NF-kappaB and CCAAA/enhancer-binding protein (C/EBP) pathways. This link includes an NF-kappaB-dependent regulation of C/EBPbeta and C/EBPdelta gene transcription and IKKi-mediated activation of C/EBP. Disruption of the NF-kappaB pathway results in the blockade of the inducible up-regulation of C/EBPbeta, C/EBPdelta, and IKKi genes. Cells lacking IKKi are normal in activation of the canonical NF-kappaB pathway but fail to induce C/EBPdelta activity and transcription of C/EBP and C/EBP-NF-kappaB target genes in response to LPS. In addition we show that, in response to LPS or tumor necrosis factor alpha, both beta and delta subunits of C/EBP interact with IKKi promoter, suggesting a feedback mechanism in the regulation of IKKi-dependent cellular processes. These data are among the first to provide insights into the biological function of IKKi. PMID- 12736253 TI - Kinetic modulation in carbonmonoxy derivatives of truncated hemoglobins: the role of distal heme pocket residues and extended apolar tunnel. AB - Truncated hemoglobins (trHbs), are a distinct and newly characterized class of small myoglobin-like proteins that are widely distributed in bacteria, unicellular eukaryotes, and higher plants. Notable and distinctive features associated with trHbs include a hydrogen-bonding network within the distal heme pocket and a long apolar tunnel linking the external solvent to the distal heme pocket. The present work compares the geminate and solvent phase rebinding kinetics from two trHbs, one from the ciliated protozoan Paramecium caudatum (P trHb) and the other from the green alga Chlamydomonas eugametos (C-trHb). Unusual kinetic patterns are observed including indications of ultrafast (picosecond) geminate rebinding of CO to C-trHb, very fast solvent phase rebinding of CO for both trHbs, time-dependent biphasic CO rebinding kinetics for P-trHb at low CO partial pressures, and for P-trHb, an increase in the geminate yield from a few percent to nearly 100% under high viscosity conditions. Species-specific differences in both the 8-ns photodissociation quantum yield and the rebinding kinetics, point to a pivotal functional role for the E11 residue. The response of the rebinding kinetics to temperature, ligand concentration, and viscosity (glycerol, trehalose) and the viscosity-dependent changes in the resonance Raman spectrum of the liganded photoproduct, together implicate both the apolar tunnel and the static and dynamic properties of the hydrogen-bonding network within the distal heme pocket in generating the unusual kinetic patterns observed for these trHbs. PMID- 12736254 TI - Enhancement of endoplasmic reticulum (ER) degradation of misfolded Null Hong Kong alpha1-antitrypsin by human ER mannosidase I. AB - Misfolded glycoproteins synthesized in the endoplasmic reticulum (ER) are degraded by cytoplasmic proteasomes, a mechanism known as ERAD (ER-associated degradation). In the present study, we demonstrate that ERAD of the misfolded genetic variant-null Hong Kong alpha1-antitrypsin is enhanced by overexpression of the ER processing alpha1,2-mannosidase (ER ManI) in HEK 293 cells, indicating the importance of ER ManI in glycoprotein quality control. We showed previously that EDEM, an enzymatically inactive mannosidase homolog, interacts with misfolded alpha1-antitrypsin and accelerates its degradation (Hosokawa, N., Wada, I., Hasegawa, K., Yorihuzi, T., Tremblay, L. O., Herscovics, A., and Nagata, K. (2001) EMBO Rep. 2, 415-422). Herein we demonstrate a combined effect of ER ManI and EDEM on ERAD of misfolded alpha1-antitrypsin. We also show that misfolded alpha1-antitrypsin NHK contains labeled Glc1Man9GlcNAc and Man5-9GlcNAc released by endo-beta-N-acetylglucosaminidase H in pulse-chase experiments with [2 3H]mannose. Overexpression of ER ManI greatly increases the formation of Man8GlcNAc, induces the formation of Glc1Man8GlcNAc and increases trimming to Man5-7GlcNAc. We propose a model whereby the misfolded glycoprotein interacts with ER ManI and with EDEM, before being recognized by downstream ERAD components. This detailed characterization of oligosaccharides associated with a misfolded glycoprotein raises the possibility that the carbohydrate recognition determinant triggering ERAD may not be restricted to Man8GlcNAc2 isomer B as previous studies have suggested. PMID- 12736255 TI - Mutation of Leu-536 in human estrogen receptor-alpha alters the coupling between ligand binding, transcription activation, and receptor conformation. AB - The estrogen receptor (ER), of which there are two forms, ERalpha and ERbeta, is a ligand-modulated transcription factor important in both normal biology and as a target for agents to prevent and treat breast cancer. Crystallographic studies of the ERalpha ligand-binding domain suggest that Leu-536 may be involved in hydrophobic interactions at the start of a helix, "helix 12," that is crucial in the agonist-stimulated activity of ERalpha, as well as in the ability of antagonists to block the activity of ERalpha. We found that certain mutations of Leu-536 increased the ligand-independent activity of ERalpha although greatly reducing or eliminating the agonist activity of 17beta-estradiol (E2) and 4 hydroxytamoxifen (4OHT), on an estrogen response element-driven and an AP-1 driven reporter. The mutations impaired the interaction of the ER ligand-binding domain with the SRC1 receptor-interacting domain in a mammalian two-hybrid system. When tested in the yeast two-hybrid system, mutation of Leu-536 increased the basal reactivity of ERalpha to probes that recognize the agonist-bound conformation but did not significantly alter its reactivity to these probes in the presence of E2. Most interestingly, mutation of Leu-536 reduced the interaction of the 4OHT-bound ERalpha and increased the reactivity of the raloxifene- or ICI 182,780-bound ERalpha, with probes that recognize the 4OHT bound ERalpha conformation in a yeast two-hybrid system. These results show that Leu-536 is critical in coupling the binding of ligand to the modulation of the conformation and activity of ERalpha. PMID- 12736256 TI - TRPC3 mediates T-cell receptor-dependent calcium entry in human T-lymphocytes. AB - Stimulation of the T-cell receptor (TCR) activates Ca2+ entry across the plasma membrane, which is a key triggering event for the T-cell-associated immune response. We show that TRPC3 channels are important for the TCR-dependent Ca2+ entry pathway. The TRPC3 gene was found to be damaged in human T-cell mutants defective in Ca2+ influx. Mutations of the TRPC3 gene were accompanied by changes of TRPC3 gene expression. Introduction of the complete human TRPC3 cDNA into those mutants rescued Ca2+ currents as well as TCR-dependent Ca2+ signals. Our data provide the initial step toward understanding the molecular nature of endogenous Ca2+ channels participating in T-cell activation and put forward TRPC3 as a new target for modulating the immune response. PMID- 12736257 TI - Furin-mediated processing of Pro-C-type natriuretic peptide. AB - C-type natriuretic peptide (CNP) is a member of the natriuretic peptide family that is involved in a variety of homeostatic processes. Here we characterize the processing essential for the conversion of the precursor, human pro-CNP, to the biologically active hormone. In human embryonic kidney 293 and chondrosarcoma SW 1353 cells, recombinant pro-CNP was converted into a mature peptide intracellularly as detected by Western analysis. Expression of recombinant human corin, a proatrial natriuretic peptide convertase, did not enhance the processing of pro-CNP in these cells. The processing of pro-CNP was inhibited in the presence of an inhibitor of the endoprotease furin but was not affected by inhibitors of matrix metalloproteinases and tumor necrosis factor-alpha convertase. In furin-deficient human colon adenocarcinoma LoVo cells, no conversion of recombinant pro-CNP to CNP was detected. Expression of recombinant human furin in LoVo cells restored the ability of these cells to process pro-CNP. Furthermore, incubation of purified recombinant human furin with LoVo cell lysate containing pro-CNP led to the conversion of the precursor to a mature peptide. The furin-processed CNP was shown to be biologically active in a cell-based cGMP assay. These results demonstrate that furin is a critical enzyme for the processing of human pro-CNP. PMID- 12736259 TI - Cross-talk between the cytosolic mevalonate and the plastidial methylerythritol phosphate pathways in tobacco bright yellow-2 cells. AB - In plants, two pathways are utilized for the synthesis of isopentenyl diphosphate, the universal precursor for isoprenoid biosynthesis. The key enzyme of the cytoplasmic mevalonic acid (MVA) pathway is 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR). Treatment of Tobacco Bright Yellow-2 (TBY-2) cells by the HMGR-specific inhibitor mevinolin led to growth reduction and induction of apparent HMGR activity, in parallel to an increase in protein representing two HMGR isozymes. Maximum induction was observed at 24 h. 1-Deoxy-d-xylulose (DX), the dephosphorylated first precursor of the plastidial 2-C-methyl-d-erythritol 4 phosphate (MEP) pathway, complemented growth inhibition by mevinolin in the low millimolar concentration range. Furthermore, DX partially re-established feedback repression of mevinolin-induced HMGR activity. Incorporation studies with [1,1,1,4-2H4]DX showed that sterols, normally derived from MVA, in the presence of mevinolin are synthesized via the MEP pathway. Fosmidomycin, an inhibitor of 1 deoxy-d-xylulose-5-phosphate reductoisomerase, the second enzyme of the MEP pathway, was utilized to study the reverse complementation. Growth inhibition by fosmidomycin of TBY-2 cells could be partially overcome by MVA. Chemical complementation was further substantiated by incorporation of [2-13C]MVA into plastoquinone, representative of plastidial isoprenoids. Best rates of incorporation of exogenous stably labeled precursors were observed in the presence of both inhibitors, thereby avoiding internal isotope dilution. PMID- 12736258 TI - X-ray crystal structure of the liver X receptor beta ligand binding domain: regulation by a histidine-tryptophan switch. AB - The x-ray crystal structures of the human liver X receptor beta ligand binding domain complexed to sterol and nonsterol agonists revealed a perpendicular histidinetryptophan switch that holds the receptor in its active conformation. Hydrogen bonding interactions with the ligand act to position the His-435 imidazole ring against the Trp-457 indole ring, allowing an electrostatic interaction that holds the AF2 helix in the active position. The neutral oxysterol 24(S),25-epoxycholesterol accepts a hydrogen bond from His-435 that positions the imidazole ring of the histidine above the pyrrole ring of the tryptophan. In contrast, the acidic T0901317 hydroxyl group makes a shorter hydrogen bond with His-435 that pulls the imidazole over the electron-rich benzene ring of the tryptophan, possibly strengthening the electrostatic interaction. Point mutagenesis of Trp-457 supports the observation that the ligand-histidine-tryptophan coupling is different between the two ligands. The lipophilic liver X receptor ligand-binding pocket is larger than the corresponding steroid hormone receptors, which allows T0901317 to adopt two distinct conformations. These results provide a molecular basis for liver X receptor activation by a wide range of endogenous neutral and acidic ligands. PMID- 12736260 TI - Mutagenesis suggests several roles of Snu114p in pre-mRNA splicing. AB - Snu114p, a yeast U5 small nuclear ribonucleoprotein (snRNP) homologous to the ribosomal GTPase EF-2, was recently found to play a part in the dissociation of U4 small nuclear RNA (snRNA) from U6 snRNA. Here, we show that purified Snu114p binds GTP specifically. To test the possibility that binding and hydrolysis of GTP by Snu114p are required to stimulate the unwinding of U4 from U6, we produced several mutations of Snu114p. Residues whose mutations led to lethal phenotypes were all clustered in the P loop and in the guanine-ring binding sequence (NKXD) of the G domain, which in elongation factor-G is required for the binding and hydrolysis of GTP. An arginine residue in domain II, which in EF-G forms a salt bridge with a residue of the G domain, when mutated in Snu114p (R487E), led to a temperature-sensitive phenotype. The substitution D271N in the NKXD sequence is predicted to bind XTP instead of GTP. Spliceosomes containing this mutant, isolated by affinity chromatography after heat treatment, retained U4 snRNA paired with the U6 snRNA. U4 snRNA was released efficiently only when these arrested spliceosomes were reactivated by lowering the temperature in the presence of a mixture of ATP and XTP. Because non-hydrolyzable XTP analogues did not consent the release of U4, we conclude that the release requires hydrolysis of XTP. This suggests that Snu114p needs GTP to influence, directly or indirectly, the unwinding of U4 from U6. An additional role for Snu114p is also demonstrated: after growth of the D271N and R487E strains at high temperatures, we observed decreased levels of the U5 and the U4/U6.U5 snRNPs. This indicates that, before splicing, Snu114p plays a part in the assembly of both particles. PMID- 12736261 TI - Circular permutation of 5-aminolevulinate synthase: effect on folding, conformational stability, and structure. AB - The first and regulatory step of heme biosynthesis in mammals begins with the pyridoxal 5'-phosphate-dependent condensation reaction catalyzed by 5 aminolevulinate synthase. The enzyme functions as a homodimer with the two active sites at the dimer interface. Previous studies demonstrated that circular permutation of 5-aminolevulinate synthase does not prevent folding of the polypeptide chain into a structure amenable to binding of the pyridoxal 5' phosphate cofactor and assembly of the two subunits into a functional enzyme. However, while maintaining a wild type-like three-dimensional structure, active, circularly permuted 5-aminolevulinate synthase variants possess different topologies. To assess whether the aminolevulinate synthase overall structure can be reached through alternative or multiple folding pathways, we investigated the guanidine hydrochloride-induced unfolding, conformational stability, and structure of active, circularly permuted variants in relation to those of the wild type enzyme using fluorescence, circular dichroism, activity, and size exclusion chromatography. Aminolevulinate synthase and circularly permuted variants folded reversibly; the equilibrium unfolding/refolding profiles were biphasic and, in all but one case, protein concentration-independent, indicating a unimolecular process with the presence of at least one stable intermediate. The formation of this intermediate was preceded by the disruption of the dimeric interface or dissociation of the dimer without significant change in the secondary structural content of the subunits. In contrast to the similar stabilities associated with the dimeric interface, the energy for the unfolding of the intermediate as well as the overall conformational stabilities varied among aminolevulinate synthase and variants. The unfolding of one functional permuted variant was protein concentration-dependent and had a potentially different folding mechanism. We propose that the order of the ALAS secondary structure elements does not determine the ability of the polypeptide chain to fold but does affect its folding mechanism. PMID- 12736262 TI - SINK is a p65-interacting negative regulator of NF-kappaB-dependent transcription. AB - The transcription factor NF-kappaB plays important roles in inflammation and cell survival. In this study, we identified SINK, an NF-kappaB-inducible protein. Overexpression of SINK inhibited NF-kappaB-dependent transcription induced by tumor necrosis factor (TNF) stimulation or its downstream signaling proteins but did not inhibit NF-kappaB translocation to the nucleus and binding to DNA. Co immunoprecipitation and in vitro kinase assays indicated that SINK specifically interacted with the NF-kappaB transactivator p65 and inhibited p65 phosphorylation by the catalytic subunit of protein kinase A, which has previously been shown to regulate NF-kappaB activation. Consistent with its role in inhibition of NF-kappaB-dependent transcription, SINK also sensitized cells to apoptosis induced by TNF and TRAIL (TNF-related apoptosis-inducing ligand). Taken together, these data suggest that SINK is critically involved in a novel negative feedback control pathway of NF-kappaB-induced gene expression. PMID- 12736263 TI - Elucidation of molecular events leading to neutrophil apoptosis following phagocytosis: cross-talk between caspase 8, reactive oxygen species, and MAPK/ERK activation. AB - Phagocytosis of complement-opsonized targets is a primary function of neutrophils at sites of inflammation, and the clearance of neutrophils that have phagocytosed microbes is important for the resolution of inflammation. Our previous work suggests that phagocytosis leads to rapid neutrophil apoptosis that is inhibited by antibody to the beta2 integrin, Mac-1, and requires NADPH oxidase-derived reactive oxygen species (ROS) generated during phagocytosis. Here we report that phagocytosis-induced cell death (PICD) does not occur in Mac-1-deficient murine neutrophils, suggesting that PICD proceeds through a bona fide Mac-1-dependent pathway. A sustained, intracellular oxidative burst is associated with PICD. Furthermore, PICD does not require traditional death receptors, Fas, or tumor necrosis factor (TNF) receptor. TNF but not Fas synergizes with phagocytosis to enhance significantly PICD by increasing the oxidative burst, and this is Mac-1 dependent. Phagocytosis-induced ROS promote cleavage/activation of caspases 8 and 3, key players in most extrinsic ("death receptor") mediated pathways of apoptosis, and caspases 8 and 3 but not caspase 9/mitochondria, are required for PICD. This suggests that ROS target the extrinsic versus the intrinsic ("stress stimulus") apoptotic pathway. Phagocytosis also triggers a competing MAPK/ERK dependent survival pathway that provides resistance to PICD likely by down regulating caspase 8 activation. The anti-apoptotic factor granulocyte-macrophage colony-stimulating factor (GM-CSF) significantly enhances ROS generation associated with phagocytosis. Despite this, it completely suppresses PICD by sustaining ERK activation and inhibiting caspase 8 activation in phagocytosing neutrophils. Together, these studies suggest that Mac-1-mediated phagocytosis promotes apoptosis through a caspase 8/3-dependent pathway that is modulated by NADPH oxidase-generated ROS and MAPK/ERK. Moreover, TNF and GM-CSF, likely encountered by phagocytosing neutrophils at inflammatory sites, exploit pro-(ROS) and anti-apoptotic (ERK) signals triggered by phagocytosis to promote or suppress PICD, respectively, and thus modulate the fate of phagocytosing neutrophils. PMID- 12736264 TI - CCHX zinc finger derivatives retain the ability to bind Zn(II) and mediate protein-DNA interactions. AB - Classical (CCHH) zinc fingers are among the most common protein domains found in eukaryotes. They function as molecular recognition elements that mediate specific contact with DNA, RNA, or other proteins and are composed of a betabetaalpha fold surrounding a single zinc ion that is ligated by two cysteine and two histidine residues. In a number of variant zinc fingers, the final histidine is not conserved, and in other unrelated zinc binding domains, residues such as aspartate can function as zinc ligands. To test whether the final histidine is required for normal folding and the DNA-binding function of classical zinc fingers, we focused on finger 3 of basic Kruppel-like factor. The structure of this domain was determined using NMR spectroscopy and found to constitute a typical classical zinc finger. We generated a panel of substitution mutants at the final histidine in this finger and found that several of the mutants retained some ability to fold in the presence of zinc. Consistent with this result, we showed that mutation of the final histidine had only a modest effect on DNA binding in the context of the full three-finger DNA-binding domain of basic Kruppel-like factor. Further, the zinc binding ability of one of the point mutants was tested and found to be indistinguishable from the wild-type domain. These results suggest that the final zinc chelating histidine is not an essential feature of classical zinc fingers and have implications for zinc finger evolution, regulation, and the design of experiments testing the functional roles of these domains. PMID- 12736265 TI - Natural disulfide bond-disrupted mutants of AVR4 of the tomato pathogen Cladosporium fulvum are sensitive to proteolysis, circumvent Cf-4-mediated resistance, but retain their chitin binding ability. AB - The extracellular AVR4 elicitor of the pathogenic fungus Cladosporium fulvum induces defense responses in the tomato genotype Cf-4. Here, the four disulfide bonds of AVR4 were identified as Cys-11-41, Cys-21-27, Cys-35-80, and Cys-57-72 by partial reduction with Tris-(2-carboxyethyl)-phosphine hydrochloride, subsequent cyanylation, and base-catalyzed chain cleavage. The resulting peptide fragments were analyzed by mass spectrometry. Sequence homology and the disulfide bond pattern revealed that AVR4 contains an invertebrate (inv) chitin-binding domain (ChBD). Binding of AVR4 to chitin was confirmed experimentally. The three disulfide bonds encompassing the inv ChBD motif are also required for protein stability of AVR4. Independent disruption of each of the three conserved disulfide bonds in AVR4 resulted in a protease-sensitive protein, whereas the fourth disulfide bond appeared not to be required for protein stability. Most strains of C. fulvum virulent on Cf-4 tomato contain Cys to Tyr substitutions in AVR4 involving two (Cys-11-41, Cys-35-80) of the three disulfide bonds present in the inv ChBD motif. These natural Cys to Tyr mutant AVR4 proteins did retain their chitin binding ability and when bound to chitin were less sensitive to proteases. Thus, the widely applied tomato Cf-4 resistance gene is circumvented by C. fulvum by amino acid substitutions affecting two disulfide bonds in AVR4 resulting in the absence of the corresponding AVR4 isoforms in apoplastic fluid. However, these natural isoforms of AVR4 appear to have retained their intrinsic function, i.e. binding to chitin present in the cell wall of C. fulvum, most likely to protect it against the deleterious effects of plant chitinases. PMID- 12736266 TI - Oligomerization, membrane anchoring, and cellulose-binding characteristics of AbpS, a receptor-like Streptomyces protein. AB - Streptomyces reticuli produces a 34.6-kDa surface-anchored protein (AbpS) whose surface-exposed N terminus binds strongly to Avicel, a dominantly crystalline type of cellulose. The generation of a large set of mutated abpS-genes and the subsequent analysis of the corresponding proteins in vitro as well as in vivo in a Streptomyces host allow the assignment of the following characteristics for AbpS. (i) Amino acid residues participating directly in the cellulose-interaction are located at the N terminus. (ii) As ascertained by cross-linking experiments, AbpS forms homotetramers in its soluble as well as cellulose-bound form. (iii) The intermolecular assembly of four AbpS molecules is governed by two domains (including amino acids 60-110 and 161-212). Both domains possess large portions of alpha-helical regions in which hydrophobic amino acids are located on one side as known from coiled-coil proteins. (iv) The C-terminal part of AbpS comprising 35 amino acids contains a transmembrane domain. Due to the surface-exposed N terminus of AbpS and the presence of transmembrane helix the C terminus has to be situated in the cytoplasm of the S. reticuli hyphae. Thus AbpS connects the interior of the mycelia with the extracellular space and binds cellulose using a unique cellulose-binding module. PMID- 12736267 TI - Proteome analysis reveals caspase activation in hyporesponsive CD4 T lymphocytes induced in vivo by the oral administration of antigen. AB - The oral administration of antigen can lead to systemic antigen-specific hyporesponsiveness, also known as oral tolerance. This phenomenon is a representative form of immune tolerance to exogenous antigen under physiological conditions. We have previously reported that long term feeding of dietary antigen to ovalbumin-specific T cell receptor (TCR) transgenic mice induced oral tolerance of peripheral T cells with impairment in their TCR-induced calcium signaling pathway. In this study, we utilized two-dimensional electrophoresis to compare intracellular protein expression patterns of orally tolerant and unsensitized CD4 T cells. We detected 26 increased and 16 decreased protein spots and identified 35 of these by mass spectrometry. The results indicated that the expression of caspases was up-regulated and that the protein levels of intact proteins susceptible to caspase cleavage, such as Grb2-related adaptor downstream of Shc (GADS), were decreased in orally tolerant CD4 T cells. Western blotting experiments confirmed that expression of the active form of caspase-3 and the antiapoptotic factor, X-linked inhibitor of apoptosis, were both up-regulated in orally tolerant CD4 T cells, which were found to be nonapoptotic. We further demonstrated that orally tolerant CD4 T cells could not form normal TCR signaling complexes associated with GADS and showed down-regulated phospholipase C-gamma1 activation, which is likely to contribute to the impairment of TCR-induced calcium signaling. Our findings indicate that orally tolerant CD4 T cells up regulate caspase activation and show decreased levels of caspase-targeted proteins, including TCR signaling-associated molecules, while up-regulating antiapoptotic factors, all of which appear to contribute to their unique tolerant characteristics. PMID- 12736268 TI - Structure and dynamics of the phospholipase C-delta1 pleckstrin homology domain located at the lipid bilayer surface. AB - Despite the importance of signal transduction pathways at membrane surfaces, there have been few means of investigating their molecular mechanisms based on the structural information of membrane-bound proteins. We applied solid state NMR as a novel method to obtain structural information about the phospholipase C delta1 (PLC-delta1) pleckstrin homology (PH) domain at the lipid bilayer surface. NMR spectra of the alanine residues in the vicinity of the beta5/beta6 loop in the PH domain revealed changes in local conformations due to the membrane localization of the protein. We propose that these conformational changes originate from a hydrophobic interaction between the amphipathic alpha-helix located in the beta5/beta6 loop and the hydrophobic layer of the membrane and contribute to the membrane binding affinity, interdomain interactions and intermolecular interactions of PLC-delta1. PMID- 12736269 TI - Activation of metabotropic glutamate receptor mGlu5 on nuclear membranes mediates intranuclear Ca2+ changes in heterologous cell types and neurons. AB - Nuclear Ca2+ plays a critical role in many cellular functions although its mode (s) of regulation is unclear. This study shows that the metabotropic glutamate receptor, mGlu5, mobilizes nuclear Ca2+ independent of cytosolic Ca2+ regulation. Immunocytochemical, ultrastructural, and subcellular fractionation techniques revealed that the metabotropic glutamate receptor, mGlu5, can be localized to nuclear membranes in heterologous cells as well as midbrain and cortical neurons. Nuclear mGlu5 receptors derived from HEK cells or cortical cell types bound [3H]quisqualate. When loaded with Oregon Green BAPTA, nuclei isolated from mGlu5 expressing HEK cells responded to the addition of glutamate with rapid, oscillatory [Ca2+] elevations that were blocked by antagonist or EGTA. In contrast, carbachol-activation of endogenous muscarinic receptors led to cytoplasmic but not nuclear Ca2+ responses. Similarly, activation of mGlu5 receptors expressed on neuronal nuclei led to sustained Ca2+ oscillatory responses. These results suggest mGlu5 may mediate intranuclear signaling pathways. PMID- 12736270 TI - Discrimination of ATP, ADP, and AMPPNP by chaperonin GroEL: hexokinase treatment revealed the exclusive role of ATP. AB - The double ring chaperonin GroEL binds unfolded protein, ATP, and GroES to the same ring, generating the cis ternary complex in which folding occurs within the cavity capped by GroES (cis folding). The functional role of ATP, however, remains unclear since several reports have indicated that ADP and AMPPNP (5' adenylyl-beta,gamma-imidodiphosphate) are also able to support the formation of the cis ternary complex and the cis folding. To minimize the effect of contaminated ATP and adenylate kinase, we have included hexokinase plus glucose in the reaction mixtures and obtained new results. In ADP and AMPPNP, GroES bound quickly to GroEL but bound very slowly to the GroEL loaded with unfolded rhodanese or malate dehydrogenase. ADP was unable to support the formation of cis ternary complex and cis folding. AMPPNP supported cis folding of malate dehydrogenase to some extent but not cis folding of rhodanese. In the absence of hexokinase, apparent cis folding of rhodanese and malate dehydrogenase was observed in ADP and AMPPNP. Thus, the exclusive role of ATP in generation of the cis ternary complex is now evident. PMID- 12736271 TI - Central role of the proteasome in senescence and survival of human fibroblasts: induction of a senescence-like phenotype upon its inhibition and resistance to stress upon its activation. AB - Normal human fibroblasts undergo a limited number of divisions in culture and progressively they reach a state of irreversible growth arrest, a process termed as replicative senescence. The proteasome is the major cellular proteolytic machinery, the function of which is impaired during replicative senescence. However, the exact causes of its malfunction in these conditions are unknown. Using WI38 fibroblasts as a model for cellular senescence we have observed reduced levels of proteasomal peptidase activities coupled with increased levels of both oxidized and ubiquitinated proteins in senescent cells. We have found the catalytic subunits of the 20 S complex and subunits of the 19 S regulatory complex to be down-regulated in senescent cells. This is accompanied by a decrease in the level of both 20 S and 26 S complexes. Partial inhibition of proteasomes in young cells caused by treatment with specific inhibitors induced a senescence-like phenotype, thus demonstrating the fundamental importance of the proteasome for retaining cellular maintenance and homeostasis. Stable overexpression of beta1 and beta5 subunits in WI38 established cell lines was shown to induce elevated expression levels of beta1 subunit in beta5 transfectants and vice versa. Transfectants possess increased proteasome activities and most importantly, increased capacity to cope better with various stresses. In summary these data demonstrate the central role of the proteasome during cellular senescence and survival as well as provide insights toward a better understanding of proteasome regulation. PMID- 12736272 TI - Thrombospondin-bound integrin-associated protein (CD47) physically and functionally modifies integrin alphaIIbbeta3 by its extracellular domain. AB - Integrin-associated protein (IAP/CD47) is a receptor for the C-terminal cell binding domain of thrombospondin (TS). A peptide from the C-terminal cell binding domain, KRFYVVMWKK (4N1K) binds to IAP and stimulates the integrin-dependent cell functions, including platelet aggregation. We investigated the mechanism by which TS-bound IAP modulates the affinity of platelet integrin, alphaIIbbeta3. Platelet aggregation induced by 4N1K was not completely inhibited by energy depletion with sodium azide and 2-deoxy-d-glucose, although ADP or collagen-induced platelet response was completely inhibited. The binding of ligand-mimetic antibody PAC1 to alphaIIbbeta3 was also induced in the energy-depleted platelets. In the transfected Namalwa cells, 4N1K induced activation of the alphaIIbbeta3 with mutated beta3 (Ser-752 to Pro), which is a non-responsive form to inside-out signaling, as well as wild type alphaIIbbeta3. The truncated form of IAP with only the extracellular immunoglobulin-like (Ig) domain was sufficient for the activation of alphaIIbbeta3 in Chinese hamster ovary cells, although the IAP mediated intracellular signaling was abolished, which was monitored by the absence of down-regulation of mitogen-activated protein kinase phosphorylation. Furthermore, the soluble recombinant Ig domain of IAP induced PAC1 binding to alphaIIbbeta3 on Chinese hamster ovary cells when added with 4N1K. Physical association between the soluble recombinant Ig domain of IAP and purified alphaIIbbeta3 was detected in the presence of 4N1K. These data indicate that the extracellular Ig domain of IAP, when bound to TS, interacts with alphaIIbbeta3 and can change alphaIIbbeta3 in a high affinity state without the requirement of intracellular signaling. This extracellular event would be a novel mechanism of affinity modulation of integrin. PMID- 12736273 TI - Coenzyme q10 prevents apoptosis by inhibiting mitochondrial depolarization independently of its free radical scavenging property. AB - The permeability transition pore (PTP) is a mitochondrial channel whose opening causes the mitochondrial membrane potential (deltapsi) collapse that leads to apoptosis. Some ubiquinone analogues have been demonstrated previously to modulate the PTP open-closed transition in isolated mitochondria and thought to act through a common PTP-binding site rather than through oxidation-reduction reactions. We have demonstrated recently both in vitro and in vivo that the ubiquitous free radical scavenger and respiratory chain coenzyme Q10 (CoQ10) prevents keratocyte apoptosis induced by excimer laser irradiation more efficiently than other antioxidants. On this basis, we hypothesized that the antiapoptotic property of CoQ10 could be independent of its free radical scavenging ability and related to direct inhibition of PTP opening. In this study, we have verified this hypothesis by evaluating the antiapoptotic effects of CoQ10 in response to apoptotic stimuli, serum starvation, antimycin A, and ceramide, which do not generate free radicals, in comparison to control, free radical-generating UVC irradiation. As hypothesized, CoQ10 dramatically reduced apoptotic cell death, attenuated ATP decrease, and hindered DNA fragmentation elicited by all apoptotic stimuli. This was accompanied by inhibition of mitochondrial depolarization, cytochrome c release, and caspase 9 activation. Because these events are consequent to mitochondrial PTP opening, we suggest that the antiapoptotic activity of CoQ10 could be related to its ability to prevent this phenomenon. PMID- 12736274 TI - Amino acids regulate hepatocyte proliferation through modulation of cyclin D1 expression. AB - The mechanisms by which amino acids regulate the cell cycle are not well characterized. In this study, we examined the control of hepatocyte proliferation by amino acids and protein intake. In short-term culture, hepatocytes demonstrated normal entry into S phase and cell cycle protein expression in the absence of essential amino acids. However, deprivation of a set of nonessential amino acids (NEAA) potently inhibited cell cycle progression and selectively down regulated the expression of proliferation-control proteins. Notably, NEAA withdrawal after the mitogen restriction point still inhibited entry into S phase, suggesting that these amino acids regulate a distinct checkpoint. Cyclin D1, an important mediator of hepatocyte proliferation, was markedly inhibited at the transcriptional level by NEAA deprivation, and transfection with cyclin D1 (but not cyclin E) overcame the cell cycle arrest. As previously shown, protein deprived mice demonstrated impaired hepatocyte proliferation in vivo after 70% partial hepatectomy. The expression of cyclin D1 and downstream cell cycle proteins after partial hepatectomy was inhibited in these mice. Transfection with cyclin D1 in vivo triggered hepatocyte DNA synthesis and the expression of S phase proteins in the absence of dietary protein. Cyclin D1 also induced global protein synthesis in NEAA-deprived hepatocytes and promoted liver growth in vivo in the setting of protein deprivation. These results indicate that cyclin D1 is a key target of amino acid signaling in hepatocytes. PMID- 12736275 TI - BACE1- and BACE2-expressing human cells: characterization of beta-amyloid precursor protein-derived catabolites, design of a novel fluorimetric assay, and identification of new in vitro inhibitors. AB - We have set up stably transfected HEK293 cells overexpressing the beta-secretases BACE1 and BACE2 either alone or in combination with wild-type beta-amyloid precursor protein (betaAPP). The characterization of the betaAPP-derived catabolites indicates that cells expressing BACEs produce less genuine Abeta1- 40/42 but higher amounts of secreted sAPPbeta and N-terminal-truncated Abeta species. This was accompanied by a concomitant modulation of the C-terminal counterpart products C89 and C79 for BACE1 and BACE2, respectively. These cells were used to set up a novel BACE assay based on two quenched fluorimetric substrates mimicking the wild-type (JMV2235) and Swedish-mutated (JMV2236) betaAPP sequences targeted by BACE activities. We show that BACEs activities are enhanced by the Swedish mutation and maximal at pH 4.5. The specificity of this double assay for genuine beta-secretase activity was demonstrated by means of cathepsin D, a "false positive" BACE candidate. Thus, cathepsin D was unable to cleave preferentially the JMV2236-mutated substrate. The selectivity of the assay was also emphasized by the lack of JMV cleavage triggered by other "secretases" candidates such as ADAM10 (A disintegrin and metalloprotease 10), tumor necrosis alpha-converting enzyme, and presenilins 1 and 2. Finally, the assay was used to screen for putative in vitro BACE inhibitors. We identified a series of statine derived sequences that dose-dependently inhibited BACE1 and BACE2 activities with IC50 in the micromolar range, some of which displaying selectivity for either BACE1 or BACE2. PMID- 12736276 TI - Probing lipid mobility of raft-exhibiting model membranes by fluorescence correlation spectroscopy. AB - Confocal fluorescence microscopy and fluorescence correlation spectroscopy (FCS) have been employed to investigate the lipid spatial and dynamic organization in giant unilamellar vesicles (GUVs) prepared from ternary mixtures of dioleoyl phosphatidylcholine/sphingomyelin/cholesterol. For a certain range of cholesterol concentration, formation of domains with raft-like properties was observed. Strikingly, the lipophilic probe 1,1'-dioctadecyl-3,3,3',3' tetramethylindocarbocyanine perchlorate (DiI-C18) was excluded from sphingomyelin enriched regions, where the raft marker ganglioside GM1 was localized. Cholesterol was shown to promote lipid segregation in dioleoyl phosphatidylcholine-enriched, liquid-disordered, and sphingomyelin-enriched, liquid-ordered phases. Most importantly, the lipid mobility in sphingomyelin enriched regions significantly increased by increasing the cholesterol concentration. These results pinpoint the key role, played by cholesterol in tuning lipid dynamics in membranes. At cholesterol concentrations >50 mol%, domains vanished and the lipid diffusion slowed down upon further addition of cholesterol. By taking the molecular diffusion coefficients as a fingerprint of membrane phase compositions, FCS is proven to evaluate domain lipid compositions. Moreover, FCS data from ternary and binary mixtures have been used to build a ternary phase diagram, which shows areas of phase coexistence, transition points, and, importantly, how lipid dynamics varies between and within phase regions. PMID- 12736277 TI - The long-QT syndrome--bedside to bench to bedside. PMID- 12736278 TI - Estrogen excess associated with novel gain-of-function mutations affecting the aromatase gene. AB - BACKGROUND: Gynecomastia of prepubertal onset may result from increased estrogen owing to excessive aromatase activity in extraglandular tissues. A gene in chromosome 15q21.2 encodes aromatase, the key enzyme for estrogen biosynthesis. Several physiologic tissue-specific promoters regulate the expression of aromatase, giving rise to messenger RNA (mRNA) species with an identical coding region but tissue-specific 5'-untranslated regions in placenta, gonads, brain, fat, and skin. METHODS: We studied skin, fat, and blood samples from a 36-year old man, his 7-year-old son, and an unrelated 17-year-old boy with severe gynecomastia of prepubertal onset and hypogonadotropic hypogonadism caused by elevated estrogen levels. RESULTS: Aromatase activity and mRNA levels in fat and skin and whole-body aromatization of androstenedione were severely elevated. Treatment with an aromatase inhibitor decreased serum estrogen levels and normalized gonadotropin and testosterone levels. The 5'-untranslated regions of aromatase mRNA contained the same sequence (FLJ) in the father and son and another sequence (TMOD3) in the unrelated boy; neither sequence was found in control subjects. These 5'-untranslated regions normally make up the first exons of two ubiquitously expressed genes clustered in chromosome 15q21.2-3 in the following order (from telomere to centromere): FLJ, TMOD3, and aromatase. The aromatase gene is normally transcribed in the direction opposite to that of TMOD3 and FLJ. Two distinct heterozygous inversions reversed the direction of the TMOD3 or FLJ promoter in the patients. CONCLUSIONS: Heterozygous inversions in chromosome 15q21.2-3, which caused the coding region of the aromatase gene to lie adjacent to constitutively active cryptic promoters that normally transcribe other genes, resulted in severe estrogen excess owing to the overexpression of aromatase in many tissues. PMID- 12736279 TI - Risk stratification in the long-QT syndrome. AB - BACKGROUND: Mutations in potassium-channel genes KCNQ1 (LQT1 locus) and KCNH2 (LQT2 locus) and the sodium-channel gene SCN5A (LQT3 locus) are the most common causes of the long-QT syndrome. We stratified risk according to the genotype, in conjunction with other clinical variables such as sex and the length of the QT interval. METHODS: We evaluated 647 patients (386 with a mutation at the LQT1 locus, 206 with a mutation at the LQT2 locus, and 55 with a mutation at the LQT3 locus) from 193 consecutively genotyped families with the long-QT syndrome. The cumulative probability of a first cardiac event, defined as the occurrence of syncope, cardiac arrest, or sudden death before the age of 40 years and before the initiation of therapy, was determined according to genotype, sex, and the QT interval corrected for heart rate (QTc). Within each genotype we also assessed risk in the four categories derived from the combination of sex and QTc (<500 msec or > or =500 msec). RESULTS: The incidence of a first cardiac event before the age of 40 years and before the initiation of therapy was lower among patients with a mutation at the LQT1 locus (30 percent) than among those with a mutation at the LQT2 locus (46 percent) or those with a mutation at the LQT3 locus (42 percent) (P<0.001 by Fisher's exact test). Multivariate analysis showed that the genetic locus and the QTc, but not sex, were independent predictors of risk. The QTc was an independent predictor of risk among patients with a mutation at the LQT1 locus and those with a mutation at the LQT2 locus but not among those with a mutation at the LQT3 locus, whereas sex was an independent predictor of events only among those with a mutation at the LQT3 locus. CONCLUSIONS: The locus of the causative mutation affects the clinical course of the long-QT syndrome and modulates the effects of the QTc and sex on clinical manifestations. We propose an approach to risk stratification based on these variables. PMID- 12736281 TI - Images in clinical medicine. Abdominal aortic aneurysm as an incidental finding. PMID- 12736280 TI - High-dose chemotherapy with hematopoietic stem-cell rescue for multiple myeloma. AB - BACKGROUND: High-dose therapy with supporting autologous stem-cell transplantation remains a controversial treatment for cancer. In multiple myeloma, first-line regimens incorporating high-dose therapy yield higher remission rates than do conventional-dose treatments, but evidence that this translates into improved survival is limited. METHODS: In this multicenter study, the Medical Research Council Myeloma VII Trial, we randomly assigned 407 patients with previously untreated multiple myeloma who were younger than 65 years of age to receive either standard conventional-dose combination chemotherapy or high dose therapy and an autologous stem-cell transplant. RESULTS: Among the 401 patients who could be evaluated, the rates of complete response were higher in the intensive-therapy group than in the standard-therapy group (44 percent vs. 8 percent, P<0.001). The rates of partial response were similar (42 percent and 40 percent, respectively; P=0.72), and the rates of minimal response were lower in the intensive-therapy group than in the standard-therapy group (3 percent vs. 18 percent, P<0.001). Intention-to-treat analysis showed a higher rate of overall survival (P=0.04 by the log-rank test) and progression-free survival (P<0.001) in the intensive-therapy group than in the standard-therapy group. As compared with standard therapy, intensive treatment increased median survival by almost 1 year (54.1 months [95 percent confidence interval, 44.9 to 65.2] vs. 42.3 months [95 percent confidence interval, 33.1 to 51.6]). There was a trend toward a greater survival benefit in the group of patients with a poor prognosis, as defined by a high beta2-microglobulin level (more than 8 mg per liter). CONCLUSIONS: High-dose therapy with autologous stem-cell rescue is an effective first-line treatment for patients with multiple myeloma who are younger than 65 years of age. PMID- 12736282 TI - Effect of a mental health "carve-out" program on the continuity of antipsychotic therapy. AB - BACKGROUND: On July 1, 1996, as a cost-containment strategy, Tennessee's expanded Medicaid program, TennCare, rapidly shifted the provision of mental health services to a fully capitated, specialty "carve-out" program, TennCare Partners. We studied the effect of this transition on the continuity of antipsychotic therapy among patients with severe mental illness who had previously adhered to treatment. METHODS: Study patients were 21 to 64 years of age, were enrolled throughout the study period, and had adhered to antipsychotic therapy during a 6 month base-line period that preceded the 12 months of study follow-up. The study population included 4507 patients whose follow-up began on the day the change was implemented (the post-transition cohort) and 3644 patients whose follow-up began one year earlier (the pretransition cohort). We compared the two cohorts in terms of the loss of continuity of antipsychotic therapy (missed treatment for more than 60 days during follow-up) and the mean number of days of antipsychotic therapy during follow-up. RESULTS: As compared with the pretransition cohort, the post-transition cohort had increased odds of loss of continuity (a multivariate odds ratio of 1.18 [95 percent confidence interval, 1.07 to 1.30], P=0.001) and a shorter mean duration of antipsychotic therapy (a mean reduction of 4.2 days [95 percent confidence interval, 1.7 to 6.7], P=0.001) during follow-up. This difference was most pronounced among high-risk patients (those requiring the administration of extended-release [depot] injections of antipsychotic medications or who had been hospitalized for psychosis) at base line, for whom continuity was most important (odds ratio for loss of continuity, 1.79 [95 percent confidence interval, 1.45 to 2.22]; P<0.001; mean reduction in the number of days of antipsychotic therapy, 14.4 days [95 percent confidence interval, 9.4 to 19.4]; P<0.001). These patients had decreased use of antipsychotic drugs immediately after the transition; the lower level persisted throughout the 12 months of follow-up. CONCLUSIONS: These findings underscore the need to ensure that shifts to widely used carve-out programs, which are designed primarily to contain costs, do not adversely affect clinical outcomes. PMID- 12736283 TI - Clinical practice. Small abdominal aortic aneurysms. PMID- 12736284 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 14-2003. A 73-year-old woman with pneumonia and progressive respiratory failure. PMID- 12736285 TI - Managing behavioral health in Medicaid. PMID- 12736286 TI - A growing network of cancer-susceptibility genes. PMID- 12736287 TI - Smallpox and smallpox vaccination. PMID- 12736288 TI - Smallpox vaccination. PMID- 12736289 TI - Multifactorial intervention and cardiovascular disease in type 2 diabetes. PMID- 12736290 TI - Serum retinol levels and fracture risk. PMID- 12736291 TI - Off-pump coronary bypass surgery. PMID- 12736292 TI - Widespread coronary inflammation in unstable angina. PMID- 12736293 TI - Easy to see but hard to find. PMID- 12736294 TI - Bromoderma after excessive ingestion of Ruby Red Squirt. PMID- 12736296 TI - The histone 3 lysine 36 methyltransferase, SET2, is involved in transcriptional elongation. AB - Existing evidence indicates that SET2, the histone 3 lysine 36 methyltransferase of Saccharomyces cerevisiae, is a transcriptional repressor. Here we show by five main lines of evidence that SET2 is involved in transcriptional elongation. First, most, if not all, subunits of the RNAP II holoenzyme co-purify with SET2. Second, all of the co-purifying RNAP II subunit, RPO21, was phosphorylated at serines 5 and 2 of the C-terminal domain (CTD) tail, indicating that the SET2 association is specific to either the elongating or SSN3 repressed forms (or both) of RNAP II. Third, the association of SET2 with CTD phosphorylated RPO21 remained in the absence of ssn3. Fourth, in the absence of ssn3, mRNA production from gal1 required SET2. Fifth, SET2 was detected on gal1 by in vivo crosslinking after, but not before, the induction of transcription. Similarly, SET2 physically associated with the transcribed region of pdr5 but was not detected on gal1 or pdr5 promoter regions. Since SET2 is also a histone methyltransferase, these results suggest a role for histone 3 lysine 36 methylation in transcriptional elongation. PMID- 12736298 TI - RNomics in Drosophila melanogaster: identification of 66 candidates for novel non messenger RNAs. AB - By generating a specialised cDNA library from four different developmental stages of Drosophila melanogaster, we have identified 66 candidates for small non messenger RNAs (snmRNAs) and have confirmed their expression by northern blot analysis. Thirteen of them were expressed at certain stages of D.melanogaster development, only. Thirty-five species belong to the class of small nucleolar RNAs (snoRNAs), divided into 15 members from the C/D subclass and 20 members from the H/ACA subclass, which mostly guide 2'-O-methylation and pseudouridylation, respectively, of rRNA and snRNAs. These also include two outstanding C/D snoRNAs, U3 and U14, both functioning as pre-rRNA chaperones. Surprisingly, the sequence of the Drosophila U14 snoRNA reflects a major change of function of this snoRNA in Diptera relative to yeast and vertebrates. Among the 22 snmRNAs lacking known sequence and structure motifs, five were located in intergenic regions, two in introns, five in untranslated regions of mRNAs, eight were derived from open reading frames, and two were transcribed opposite to an intron. Interestingly, detection of two RNA species from this group implies that certain snmRNA species are processed from alternatively spliced pre-mRNAs. Surprisingly, a few snmRNA sequences could not be found on the published D.melanogaster genome, which might suggest that more snmRNA genes (as well as mRNAs) are hidden in unsequenced regions of the genome. PMID- 12736295 TI - Unusual DNA duplex and hairpin motifs. AB - Single-stranded DNA or double-stranded DNA has the potential to adopt a wide variety of unusual duplex and hairpin motifs in the presence (trans) or absence (cis) of ligands. Several principles for the formation of those unusual structures have been established through the observation of a number of recurring structural motifs associated with different sequences. These include: (i) internal loops of consecutive mismatches can occur in a B-DNA duplex when sheared base pairs are adjacent to each other to confer extensive cross- and intra-strand base stacking; (ii) interdigitated (zipper-like) duplex structures form instead when sheared G*A base pairs are separated by one or two pairs of purine*purine mismatches; (iii) stacking is not restricted to base, deoxyribose also exhibits the potential to do so; (iv) canonical G*C or A.T base pairs are flexible enough to exhibit considerable changes from the regular H-bonded conformation. The paired bases become stacked when bracketed by sheared G.A base pairs, or become extruded out and perpendicular to their neighboring bases in the presence of interacting drugs; (v) the purine-rich and pyrimidine-rich loop structures are notably different in nature. The purine-rich loops form compact triloop structures closed by a sheared G*A, A*A, A*C or sheared-like G(anti)*C(syn) base pair that is stacked by a single residue. On the other hand, the pyrimidine-rich loops with a thymidine in the first position exhibit no base pairing but are characterized by the folding of the thymidine residue into the minor groove to form a compact loop structure. Identification of such diverse duplex or hairpin motifs greatly enlarges the repertoire for unusual DNA structural formation. PMID- 12736297 TI - The capacity to form H-DNA cannot substitute for GAGA factor binding to a (CT)n*(GA)n regulatory site. AB - Previous studies of the Drosophila melanogaster hsp26 gene promoter have demonstrated the importance of a homopurine*homopyrimidine segment [primarily (CT)n*(GA)n] for chromatin structure formation and gene activation. (CT)n regions are known to bind GAGA factor, a dominant enhancer of PEV thought to play a role in generating an accessible chromatin structure. The (CT)n region can also form an H-DNA structure in vitro under acidic pH and negative supercoiling; a detailed map of that structure is reported here. To test whether the (CT)n sequence can function through H-DNA in vivo, we have analyzed a series of hsp26-lacZ transgenes with altered sequences in this region. The results indicate that a 25 bp mirror repeat within the homopurine.homopyrimidine region, while adequate for H-DNA formation, is neither necessary nor sufficient for positive regulation of hsp26 when GAGA factor-binding sites have been eliminated. The ability to form H DNA cannot substitute for GAGA factor binding to the (CT)n sequence. PMID- 12736299 TI - Translational control of Scamper expression via a cell-specific internal ribosome entry site. AB - The mRNA of Scamper, a putative intracellular calcium channel activated by sphingosylphosphocholine, contains a long 5' transcript leader with several upstream AUGs. In this work we have investigated the role this sequence plays in the translational control of Scamper expression. The cytosolic transcription machinery of a T7 RNA polymerase recombinant vaccinia virus was used to avoid artifacts arising from cryptic promoters or mRNA processing. Based on transient transfection experiments of dicistronic and bi-monocistronic plasmids expressing reporter genes, we present evidence that the 5' transcript leader of Scamper contains a functional internal ribosome entry site (IRES). Our data indicate that Scamper translation in Madin-Darby canine kidney cells is driven by a cap independent mechanism supported by the IRES activity of its mRNA. Finally, the Scamper IRES appears to be the first IRES with specificity for kidney epithelial cells. PMID- 12736300 TI - Synthesis, thermal stability and resistance to enzymatic hydrolysis of the oligonucleotides containing 5-(N-aminohexyl)carbamoyl-2'-O-methyluridines. AB - The synthesis of oligonucleotides (ODNs) containing 5-(N-aminohexyl)carbamoyl-2' O-methyluridine (D) is described, and thermal stability and resistance to enzymatic hydrolysis of the ODNs are compared with ODNs containing 5-(N aminohexyl)carbamoyl-2'-deoxyuridine (H). The ODNs containing D and the complementary RNA demonstrated a duplex thermal stabilization of 0.4-3.9 degrees C per modification depending on the position and the number, while the ODNs containing H with the RNA showed slightly less effective thermal stabilization. Further more, the ODNs containing D were found to be more resistant to nucleolytic hydrolysis, not only by snake venom phosphodiesterase (SVPD; a 3' exonuclease) but also by DNase I (an endonuclease). The half-life of the 17mer containing five molecules of D against nucleolytic hydrolysis by SVPD was 240 times greater than the unmodified 17mer ODN, which is 1.8 times greater than the ODN containing 5Hs in the same sequence. Against DNase I, the same ODN containing 5Ds was 24 times greater stable than the unmodified 17mer and 15 times more stable than the ODN containing 5Hs. We also examined whether the duplexes formed by the ODNs containing D and the complementary RNAs could be a substrate of Escherichia coli RNase H. It was revealed that a minimum of five contiguous unmodified 2'-deoxyribonucleosides between Ds was required to constitute a substrate of E.coli RNase H. Thus, the ODN with Ds and at least five contiguous unmodified 2'-deoxyribonucleosides between Ds was found to be a candidate for a novel antisense molecule. PMID- 12736301 TI - RRP20, a component of the 90S preribosome, is required for pre-18S rRNA processing in Saccharomyces cerevisiae. AB - A strain of Saccharomyces cerevisiae, defective in small subunit ribosomal RNA processing, has a mutation in YOR145c ORF that converts Gly235 to Asp. Yor145c is a nucleolar protein required for cell viability and has been reported recently to be present in 90S pre-ribosomal particles. The Gly235Asp mutation in YOR145c is found in a KH-type RNA-binding domain and causes a marked deficiency in 18S rRNA production. Detailed studies by northern blotting and primer extension analyses show that the mutant strain impairs the early pre-rRNA processing cleavage essentially at sites A1 and A2, leading to accumulation of a 22S dead-end processing product that is found in only a few rRNA processing mutants. Furthermore, U3, U14, snR10 and snR30 snoRNAs, involved in early pre-rRNA cleavages, are not destabilized by the YOR145c mutation. As the protein encoded by YOR145c is found in pre-ribosomal particles and the mutant strain is defective in ribosomal RNA processing, we have renamed it as RRP20. PMID- 12736303 TI - Conservation of human alternative splice events in mouse. AB - Human and mouse genomes share similar long-range sequence organization, and have most of their genes being homologous. As alternative splicing is a frequent and important aspect of gene regulation, it is of interest to assess the level of conservation of alternative splicing. We examined mouse transcript data sets (EST and mRNA) for the presence of transcripts that both make spliced-alignment with the draft mouse genome sequence and demonstrate conservation of human transcript confirmed alternative and constitutive splice junctions. This revealed 15% of alternative and 67% of constitutive splice junctions as conserved; however, these numbers are patently dependent on the extent of transcript coverage. Transcript coverage of conserved splice patterns is found to correlate well between human and mouse. A model, which extrapolates from observed levels of conservation at increasing levels of transcript support, estimates overall conservation of 61% of alternative and 74% of constitutive splice junctions, albeit with broad confidence intervals. Observed numbers of conserved alternative splicing events agreed with those expected on the basis of the model. Thus, it is apparent that many, and probably most, alternative splicing events are conserved between human and mouse. This, combined with the preservation of alternative frame stop codons in conserved frame breaking events, indicates a high level of commonality in patterns of gene expression between these two species. PMID- 12736302 TI - Isolation and analyses of genes preferentially expressed during early cotton fiber development by subtractive PCR and cDNA array. AB - Cotton fibers are differentiated epidermal cells originating from the outer integuments of the ovule. To identify genes involved in cotton fiber elongation, we performed subtractive PCR using cDNA prepared from 10 days post anthesis (d.p.a.) wild-type cotton fiber as tester and cDNA from a fuzzless-lintless (fl) mutant as driver. We recovered 280 independent cDNA fragments including most of the previously published cotton fiber-related genes. cDNA macroarrays showed that 172 genes were significantly up-regulated in elongating cotton fibers as confirmed by in situ hybridization in representative cases. Twenty-nine cDNAs, including a putative vacuolar (H+)-ATPase catalytic subunit, a kinesin-like calmodulin binding protein, several arabinogalactan proteins and key enzymes involved in long chain fatty acid biosynthesis, accumulated to greater than 50 fold in 10 d.p.a. fiber cells when compared to that in 0 d.p.a. ovules. Various upstream pathways, such as auxin signal transduction, the MAPK pathway and profilin- and expansin-induced cell wall loosening, were also activated during the fast fiber elongation period. This report constitutes the first systematic analysis of genes involved in cotton fiber development. Our results suggest that a concerted mechanism involving multiple cellular pathways is responsible for cotton fiber elongation. PMID- 12736305 TI - Spectroscopic studies of the multiple binding modes of a trimethine-bridged cyanine dye with DNA. AB - The interaction between DNA and a benzothiazole-quinoline cyanine dye with a trimethine bridge (TO-PRO-3) results in the formation of three noncovalent complexes. Unbound TO-PRO-3 has an absorption maximum (lambda(max)) of 632 nm, while the bound dyes (with calf thymus DNA) have electronic transitions with lambda(max) = 514 nm (complex I), 584 nm (complex II) and 642 nm (complex III). The blue shifts in the electronic transitions and the bisignate shape of the circular dichroism bands indicate that TO-PRO-3 aggregates with DNA. Complex I has a high dye:base pair stoichiometry, which does not depend on base sequence or base modifications. The bound dyes exhibit strong interdye coupling, based on studies with a short oligonucleotide and on enhanced resonance scattering. From thermal dissociation studies, the complex is weakly associated with DNA. Studies with poly(dGdC)2 and poly(dIdC)2 and competitive binding with distamycin demonstrate that complex II is bound in the minor groove. This complex stabilizes the helix against dissociation. For complex III, the slightly red-shifted electronic transition and the stoichiometry are most consistent with intercalation. Using poly(dAdT)2, the complexes have the following dye mole fractions (X(dye)): X(dye) = 0.65 (complex I), 0.425 (complex II) and 0.34 (complex III). PMID- 12736304 TI - Limited microsynteny between the genomes of Pristionchus pacificus and Caenorhabditis elegans. AB - Nematodes are an attractive group of organisms for studying the evolution of developmental processes. Pristionchus pacificus was established as a satellite organism for comparing vulva development and other processes to Caenorhabditis elegans. The generation of a genetic linkage map of P.pacificus has provided a first insight into the structure and organization of the genome of this species. Pristionchus pacificus and C.elegans are separated from one another by >100 000 000 years such that the structure of the genomes of these two nematodes might differ substantially. To evaluate the amount of synteny between the two genomes, we have obtained 126 kb of continuous genomic sequence of P.pacificus, flanking the developmental patterning gene pal-1. Of the 20 predicted open reading frames in this interval, 11 have C.elegans orthologs. Ten of these 11 orthologs are located on C.elegans chromosome III, indicating the existence of synteny. However, most of these genes are distributed over a 12 Mb interval of the C.elegans genome and only three pairs of genes show microsynteny. Thus, intrachromosomal rearrange ments occur frequently in nematodes, limiting the likelihood of identifying orthologous genes of P.pacificus and C.elegans based on positional information within the two genomes. PMID- 12736306 TI - Mutagenic effects of 2-hydroxy-dATP on replication in a HeLa extract: induction of substitution and deletion mutations. AB - The mutagenicity of an oxidized form of dATP, 2-hydroxydeoxyadenosine 5' triphosphate (2-OH-dATP), was examined using an SV40 origin-dependent in vitro replication system with a HeLa extract. 2-OH-dATP induced mutations in a dose dependent manner and elicited substitution and deletion mutations. Of the substitutions, a G.C-->A.T transition including a tandem (CC-->TT) mutation was mainly observed. This result agrees with our previous observation that mammalian DNA polymerase alpha misincorporates the oxidized nucleotide opposite C, but is in contrast to the finding that 2-OH-dATP elicits G.C-->T.A transversions in Escherichia coli. This type of mutation was also elicited, but to a lesser extent. Interestingly, the mutagenicity of 2-OH-dATP was enhanced in the presence of 2-hydroxydeoxyadenosine 5'-diphosphate, an inhibitor of the MTH1 protein, suggesting that this protein functions in the hydrolysis of 2-OH-dATP in the replication reaction mixture, and probably in living cells. These results indicate that 2-OH-dATP is mutagenic and that its mutagenicity is suppressed by the MTH1 protein in mammalian cells. PMID- 12736307 TI - Multiple recombination pathways for sister chromatid exchange in Saccharomyces cerevisiae: role of RAD1 and the RAD52 epistasis group genes. AB - Sister chromatid exchange (SCE) can occur by several recombination mechanisms, including those directly initiated by double-strand breaks (DSBs), such as gap repair and break-induced replication (BIR), and those initiated when DNA polymerases stall, such as template switching. To elucidate SCE recombination mechanisms, we determined whether spontaneous and DNA damage-associated SCE requires specific genes within the RAD52 and RAD3 epistasis groups in Saccharomyces cerevisiae strains containing two his3 fragments, his3-Delta5' and his3-Delta3'::HOcs. SCE frequencies were measured after cells were exposed to UV, X-rays, 4-nitroquinoline 1-oxide (4-NQO) and methyl methanesulfonate (MMS), or when an HO endonuclease-induced DSB was introduced at his3-Delta3'::HOcs. Our data indicate that genes involved in gap repair, such as RAD55, RAD57 and RAD54, are required for DNA damage-associated SCE but not for spontaneous SCE. RAD50 and RAD59, genes required for BIR, are required for X-ray-associated SCE but not for SCE stimulated by HO-induced DSBs. In comparison with wild type, rates of spontaneous SCE are 10-fold lower in rad51 rad1 but not in either rad51 rad50 or rad51 rad59 double mutants. We propose that gap repair mechanisms are important in DNA damage-associated recombination, whereas alternative pathways, including a template switch pathway, play a role in spontaneous SCE. PMID- 12736309 TI - Zinc-dependent cleavage in the catalytic core of the hammerhead ribozyme: evidence for a pH-dependent conformational change. AB - We have characterized a novel Zn2+-catalyzed cleavage site between nucleotides C3 and U4 in the catalytic core of the hammerhead ribozyme. In contrast to previously described divalent metal-ion-dependent cleavage of RNA, U4 cleavage is only observed in the presence of Zn2+. This new cleavage site has an unusual pH dependence, in that U4 cleavage products are only observed above pH 7.9 and reach a maximum yield at about pH 8.5. These data, together with the fact that no metal ion-binding site is observed in proximity to the U4 cleavage site in either of the crystal structures, point toward a pH-dependent conformational change in the hammerhead ribozyme. We have described previously Zn2+-dependent cleavage between G8 and A9 in the hammerhead ribozyme and have discovered that U4 cleavage occurs only after A9 cleavage. To our knowledge, this is the first example of sequential cleavage events as a possible regulatory mechanism in ribozymes. PMID- 12736308 TI - Developmental co-variation of RNA editing extent of plastid editing sites exhibiting similar cis-elements. AB - In tobacco, 30 of 34 sites in chloroplast transcripts that undergo C-to-U RNA editing can be grouped into clusters of 2-5 sites based on sequence similarities immediately 5' to the edited C. According to a previous transgenic analysis, overexpression of transcripts representing one cluster member results in reduction in editing of all cluster members, suggesting that members of an individual cluster share a trans-factor that is present in limiting amounts. To compare leaves and roots, we quantified the editing extent at 34 sites in wild type tobacco and at three sites in spinach and Arabidopsis. We observed that transcripts of most NADH dehydrogenase subunits are edited inefficiently in roots. With few exceptions, members of the same editing site cluster co-varied in editing extent in chloroplasts versus non-green root plastids, with members of most clusters uniformly exhibiting either a high or low editing extent in roots. The start codon of the ndhD transcript must be created by editing, but the C target is edited inefficiently in roots, and no NDH-D protein could be detected upon immunoblotting. Our data are consistent with the hypothesis that cluster specific trans-factors exist and that some are less abundant in roots, limiting the editing extent of certain sites in root plastids. PMID- 12736311 TI - The solution structure of an essential stem-loop of human telomerase RNA. AB - The ribonucleoprotein enzyme telomerase maintains chromosome ends in most eukaryotes and is critical for a cell's genetic stability and its proliferative viability. All telomerases contain a catalytic protein component homologous to viral reverse transcriptases (TERT) and an RNA (TR) that provides the template sequence as well as a scaffold for ribonucleoprotein assembly. Vertebrate telomerase RNAs have three essential domains: the template, activation and stability domains. Here we report the NMR structure of an essential RNA element derived from the human telomerase RNA activation domain. The sequence forms a stem-loop structure stabilized by a GU wobble pair formed by two of the five unpaired residues capping a short double helical region. The remaining three loop residues are in a well-defined conformation and form phosphate-base stacking interactions reminiscent of other RNA loop structures. Mutations of these unpaired nucleotides abolish enzymatic activity. The structure rationalizes a number of biochemical observations, and allows us to propose how the loop may function in the telomerase catalytic cycle. The pre-formed structure of the loop exposes the bases of these three essential nucleotides and positions them to interact with other RNA sequences within TR, with the reverse transcriptase or with the newly synthesized telomeric DNA strand. The functional role of this stem loop appears to be conserved in even distantly related organisms such as yeast and ciliates. PMID- 12736310 TI - The high diversity of snoRNAs in plants: identification and comparative study of 120 snoRNA genes from Oryza sativa. AB - Using a powerful computer-assisted analysis strategy, a large-scale search of small nucleolar RNA (snoRNA) genes in the recently released draft sequence of the rice genome was carried out. This analysis identified 120 different box C/D snoRNA genes with a total of 346 gene variants, which were predicted to guide 135 2'-O-ribose methylation sites in rice rRNAs. Though not exhaustive, this analysis has revealed that rice has the highest number of known box C/D snoRNAs among eukaryotes. Interestingly, although many snoRNA genes are conserved between rice and Arabidopsis, almost half of the identified snoRNA genes are rice specific, which may highlight further the differences in rRNA methylation patterns between monocotyledons and dicotyledons. In addition to 76 singletons, 70 clusters involving 270 snoRNA genes were also found in rice. The large number of the novel snoRNA polycistrons found in the introns of rice protein-coding genes is in contrast to the one-snoRNA-per-intron organization of vertebrates and yeast, and of Arabidopsis in which only a few intronic snoRNA gene clusters were identified. Furthermore, due to a high degree of gene duplication, rice snoRNA genes are clearly redundant and exhibit great sequence variation among isoforms, allowing generation of new snoRNAs for selection. Thus, the large snoRNA gene family in plants can serve as an excellent model for a rapid and functional evolution. PMID- 12736312 TI - Solution structure of the ActD-5'-CCGTT3GTGG-3' complex: drug interaction with tandem G.T mismatches and hairpin loop backbone. AB - Binding of actinomycin D (ActD) to the seemingly single-stranded DNA (ssDNA) oligomer 5'-CCGTT3 GTGG-3' has been studied in solution using high-resolution nuclear magnetic resonance (NMR) techniques. A strong binding constant (8 x 10(6) M(-1)) and high quality NMR spectra have allowed us to determine the initial DNA structure using distance geometry as well as the final ActD-5'-CCGTT3 GTGG-3' complex structure using constrained molecular dynamics calculations. The DNA oligomer 5'-CCGTT3GTGG-3' in the complex forms a hairpin structure with tandem G.T mismatches at the stem region next to a loop of three stacked thymine bases pointing toward the major groove. Bipartite T2O-GH1 and T2O-G2NH2 hydrogen bonds were detected for the G.T mismatches that further stabilize this unusual DNA hairpin. The phenoxazone chromophore of ActD intercalates nicely between the tandem G.T mismatches in essentially one major orientation. Additional hydrophobic interactions between the ActD quinoid amino acid residues with the loop T5-T6-T7 backbone protons were also observed. The hydrophobic G-phenoxazone G interaction in the ActD-5'-CCGTT3GTGG-3' complex is more robust than that of the classical ActD- 5'-CCGCT3GCGG-3' complex, consistent with the roughly 2-fold stronger binding of ActD to the 5'-CCGTT3GTGG-3' sequence than to its 5'-CCG CT3GCGG-3' counterpart. Stabilization by ActD of a hairpin containing non canonical stem base pairs further strengthens the notion that ActD or other related compounds may serve as a sequence- specific ssDNA-binding agent that inhibits human immunodeficiency virus (HIV) and other retroviruses replicating through ssDNA intermediates. PMID- 12736313 TI - Incorporation of reporter molecule-labeled nucleotides by DNA polymerases. I. Chemical synthesis of various reporter group-labeled 2'-deoxyribonucleoside-5' triphosphates. AB - Fluorescent-labeled DNA is generated through enzymatic incorporation of fluorophore-linked 2'-deoxyribonucleoside-5'-triphosphates (dNTPs) by DNA polymerases. We describe the synthesis of a variety of dye-labeled dNTPs. Amino linker-modified 5'-triphosphates of all four naturally occurring nucleobases were used as precursors. Commercially available dyes were coupled to the amino function of the side chain. In addition, we attached novel fluorophore derivatives. The labeled products were obtained in at least 96% purity after HPLC purification. Enzymatic incorporation into DNA and subsequent extension of the modified DNA chain were studied. Vent(R) exo- DNA polymerase and a defined template-primer system were used to analyze each dye-labeled dNTP derivative. Our data suggest that the incorporation efficiency depends on the selected dye, the nucleobase or a combination of both. PMID- 12736314 TI - Incorporation of reporter molecule-labeled nucleotides by DNA polymerases. II. High-density labeling of natural DNA. AB - The modification of nucleic acids using nucleotides linked to detectable reporter or functional groups is an important experimental tool in modern molecular biology. This enhances DNA or RNA detection as well as expanding the catalytic repertoire of nucleic acids. Here we present the evaluation of a broad range of modified deoxyribonucleoside 5'-triphosphates (dNTPs) covering all four naturally occurring nucleobases for potential use in DNA modification. A total of 30 modified dNTPs with either fluorescent or non-fluorescent reporter group attachments were systematically evaluated individually and in combinations for high-density incorporation using different model and natural DNA templates. Furthermore, we show a side-by-side comparison of the incorporation efficiencies of a family A (Taq) and B (Vent(R) exo-) type DNA polymerase using the differently modified dNTP substrates. Our results show superior performance by a family B-type DNA polymerase, Vent(R) exo-, which is able to fully synthesize a 300 bp DNA product when all natural dNTPs are completely replaced by their biotin labeled dNTP analogs. Moreover, we present systematic testing of various combinations of fluorescent dye-modified dNTPs enabling the simultaneous labeling of DNA with up to four differently modified dNTPs. PMID- 12736315 TI - An assessment of three dinucleotide parameters to predict DNA curvature by quantitative comparison with experimental data. AB - Curved DNA fragments are often found near functionally important sites such as promoters and origins of replication, and hence sequence-dependent DNA curvature prediction is of great utility in genomics and bioinformatics. In light of this, an assessment of three different dinucleotide step parameters (based on gel retardation as well as crystal structure data) is carried out. These parameters (BMHT, LB and CS) are evaluated quantitatively for their ability to predict correctly the experimental results of a large set of nucleic acid sequences containing A-tracts as well as GC-rich motifs. This set contained around 40 synthetic as well as natural sequences whose solution properties have been well characterized experimentally. All three models could account reasonably well for curvature in the various DNA sequences. The CS model, where dinucleotide parameters are calculated from crystal structure data, consistently shows slightly better correlation with experimental data. Our simple analysis also indicates that presently available trinucleotide parameters fail to predict curvature in some of the well-characterized sequences. The study shows that the dinucleotide parameters with some further refinement can be used to predict sequence-dependent curvature correctly in genomic sequences. PMID- 12736316 TI - Transcription affects formation and processing of intermediates in oligonucleotide-mediated gene alteration. AB - The role of transcription in oligonucleotide (ODN)-directed gene modification has been investigated in mammalian cells. The importance of transcription is demonstrated using mammalian cell lines with varying degrees of transcription of the mutant LacZ reporter gene, residing in both episome and chromosome. Gene correction occurs more efficiently when the target gene is actively transcribed and antisense ODN is more active than sense ODN. Using an approach that combines biochemical studies with a cell-based assay to measure the functional activity of intermediates it is shown that a joint molecule, consisting of supercoiled DNA and homologous ODN targeted to correct the mutated base, is a functional intermediate in the gene repair process. Furthermore, this approach showed that a resected joint molecule is a downstream intermediate of the D-loop. These results indicate that the primary reason for efficient gene repair exhibited by the antisense ODN is its increased accessibility to the non-transcribed strand, and as a consequence an increased formation of intermediate during active transcription. Moreover, the processing of intermediates was also affected by transcription, suggesting that ODN-directed gene repair may be linked to transcription-coupled repair. Thus, transcription plays an important role in ODN directed gene repair by affecting the formation and processing of key intermediates. PMID- 12736317 TI - A crystallographic study of the binding of 13 metal ions to two related RNA duplexes. AB - Metal ions, and magnesium in particular, are known to be involved in RNA folding by stabilizing secondary and tertiary structures, and, as cofactors, in RNA enzymatic activity. We have conducted a systematic crystallographic analysis of cation binding to the duplex form of the HIV-1 RNA dimerization initiation site for the subtype-A and -B natural sequences. Eleven ions (K+, Pb2+, Mn2+, Ba2+, Ca2+, Cd2+, Sr2+, Zn2+, Co2+, Au3+ and Pt4+) and two hexammines [Co (NH3)6]3+ and [Ru (NH3)6]3+ were found to bind to the DIS duplex structure. Although the two sequences are very similar, strong differences were found in their cation binding properties. Divalent cations bind almost exclusively, as Mg2+, at 'Hoogsteen' sites of guanine residues, with a cation-dependent affinity for each site. Notably, a given cation can have very different affinities for a priori equivalent sites within the same molecule. Surprisingly, none of the two hexammines used were able to efficiently replace hexahydrated magnesium. Instead, [Co (NH3)4]3+ was seen bound by inner-sphere coordination to the RNA. This raises some questions about the practical use of [Co (NH3)6]3+ as a [Mg (H2O)6]2+ mimetic. Also very unexpected was the binding of the small Au3+ cation exactly between the Watson-Crick sites of a G-C base pair after an obligatory deprotonation of N1 of the guanine base. This extensive study of metal ion binding using X-ray crystallography significantly enriches our knowledge on the binding of middleweight or heavy metal ions to RNA, particularly compared with magnesium. PMID- 12736318 TI - The solution structure of a DNA*RNA duplex containing 5-propynyl U and C; comparison with 5-Me modifications. AB - The addition of the propynyl group at the 5 position of pyrimidine nucleotides is highly stabilising. We have determined the thermodynamic stability of the DNA.RNA hybrid r(GAAGAGAAGC)*d(GC(p)U(p)U(p)C(p)U(p) C(p)U(p)U(p)C) where p is the propynyl group at the 5 position and compared it with that of the unmodified duplex and the effects of methyl substitutions. The incorporation of the propyne group at the 5 position gives rise to a very large stabilisation of the hybrid duplex compared with the analogous 5-Me modification. The duplexes have been characterised by gel electrophoresis and NMR spectroscopy, which indicate that methyl substitutions have a smaller influence on local and global conformation than the propynyl groups. The increased NMR spectral dispersion of the propyne modified duplex allowed a larger number of experimental restraints to be measured. Restrained molecular dynamics in a fully solvated system showed that the propyne modification leads to substantial conformational rearrangements stabilising a more A-like structure. The propynyl groups occupy a large part of the major groove and make favourable van der Waals interactions with their nearest neighbours and the atoms of the rings. This enhanced overlap may account at least in part for the increased thermodynamic stability. Furthermore, the simulations show a spine of hydration in the major groove as well as in the minor groove involving the RNA hydroxyl groups. PMID- 12736319 TI - The (52-96) C-terminal domain of Vpr stimulates HIV-1 IN-mediated homologous strand transfer of mini-viral DNA. AB - Viral integrase (IN) and Vpr are both components of the human immunodeficiency virus type 1 (HIV-1) pre-integration complex. To investigate whether these proteins interact within this complex, we investigated the effects of Vpr and its subdomains on IN activity in vitro. When a 21mer oligonucleotide was used as a donor and acceptor, both Vpr and its C-terminal DNA-binding domain [(52-96)Vpr] inhibited the integration reaction, whereas the (1-51)Vpr domain did not affect IN activity. Steady-state fluorescence anisotropy showed that both full-length and (52-96)Vpr bind to the short oligonucleotide, thereby extending previous observations with long DNA. The concentrations of the two proteins required to inhibit IN activity were consistent with their affinities for the oligonucleotide. The use of a 492 bp mini-viral substrate confirmed that Vpr can inhibit the IN-mediated reaction. However, the activity of (52-96)Vpr differed notably since it stimulated specifically integration events involving two homologous mini-viral DNAs. Order of addition experiments indicated that the stimulation was maximal when IN, (50-96)Vpr and the mini-viral DNA were allowed to form a complex. Furthermore, in the presence of (50-96)Vpr, the binding of IN to the mini-viral DNA was dramatically enhanced. Taken together, these data suggest that (52-96)Vpr stimulates the formation of a specific complex between IN and the mini-viral DNA. PMID- 12736320 TI - Immunofluorescence assay and flow-cytometry selection of bead-bound aptamers. AB - An immunofluorescence assay was developed to identify proteins specifically binding to oligonucleoside phosphorodithioate (ODN) aptamers from a bead-bound ODN library. Accordingly, NF-kappaB p50 protein was incubated with either bead bound NF-kappaB consensus sequence or a bead-bound ODN combinatorial library and adsorption was then assessed using a specific primary antibody and a secondary antibody conjugated with Alexa 488 fluorescent dye. This assay avoids any problems related to fluorescently labeling target proteins. The method is straightforward and readily applicable to other transcription factors and proteins, and the feasibility of its application for high-throughput screening of large aptamer bead-based libraries by flow cytometry is demonstrated. PMID- 12736321 TI - A new and versatile method for the successful conversion of AFLP markers into simple single locus markers. AB - Genetic markers can efficiently be obtained by using amplified fragment length polymorphism (AFLP) fingerprinting because no prior information on DNA sequence is required. However, the conversion of AFLP markers from complex fingerprints into simple single locus assays is perceived as problematic because DNA sequence information is required for the design of new locus-specific PCR primers. In addition, single locus polymorphism (SNP) information is required to design an allele-specific assay. This paper describes a new and versatile method for the conversion of AFLP markers into simple assays. The protocol presented in this paper offers solutions for frequently occurring pitfalls and describes a procedure for the identification of the SNP responsible for the AFLP. By following this approach, a high success rate for the conversion of AFLP markers into locus-specific markers was obtained. PMID- 12736322 TI - Real-time PCR-based method for the estimation of genome sizes. AB - The fast and reliable estimation of the genome sizes of various species would allow for a systematic analysis of many organisms and could reveal insights into evolutionary processes. Many methods for the estimation of genome sizes have already been described. The classical methods are based on the determination of the phosphate content in the DNA backbone of total DNA isolated from a defined number of cells or on reassociation kinetics of high molecular weight genomic DNA (c(0)t assay). More recent techniques employ DNA-specific fluorescent dyes in flow cytometry analysis, image analysis or absorption cytometry after Feulgen staining. The method presented here is based on the absolute quantification of genetic elements in a known amount (mass) of genomic DNA by real-time quantitative PCR. The method was evaluated on three different eukaryotic species, Saccharomyces cerevisiae (12.1 Mb), Xiphophorus maculatus (550 Mb) and Homo sapiens sapiens (2.9 Gb), and found to be fast, highly accurate and reliable. PMID- 12736323 TI - An inbred 129SvEv GFPCre transgenic mouse that deletes loxP-flanked genes in all tissues. AB - A common method for generating mice with subtle genetic manipulations uses homologous recombination (HR) in embryonic stem (ES) cells to replace a wild-type gene with a slightly modified one. Generally, a drug resistance gene is inserted with the modified gene to select correctly targeted clones. Often, however, the presence of this drug resistance gene interferes with the normal locus and creates a null or hypomorphic allele. Flanking of the selectable marker by loxP sites followed by Cre-mediated deletion after drug selection can overcome this problem. The simplest method used to remove a loxP-flanked selectable marker is to breed an animal carrying a loxP-flanked drug resistance gene to an animal that expresses Cre recombinase in the germline. To date only outbred transgenic mice are available for this purpose. This can be problematic for phenotypic analysis in many organ systems, including the brain, and for the analysis of behavior. While attempting to make 129S6/SvEvTac inbred background (isogenic to our ES cells) mice that express Cre under the control of several tissue-specific promoters, we serendipitously generated a line that excises loxP-flanked drug resistance genes in all tissues, including the germline. This reagent allows deletion of loxP-flanked sequences while maintaining the mutation on an inbred background. PMID- 12736325 TI - A physiological correlate of the "Zoom Lens" of visual attention. AB - Attending a certain region in space enhances activity in visual areas retinotopically mapped to this region; stimuli presented in this region are preferentially processed. The zoom lens model of visual attention proposes that the attended region can be adjusted in size and predicts a tradeoff between its size and processing efficiency because of limited processing capacities. By means of event-related functional magnetic resonance imaging, we analyzed neural activity in multiple visual areas as a function of the size of an attended visual field region, which was defined by a spatial cue stimulus. After cueing, a target object, defined by a specific feature conjunction, had to be identified among objects within the cued region. Neural activity preceding the objects in multiple retinotopic visual areas correlated with the size of the attended region, as did subjects' performance. While the extent of activated retinotopic visual cortex increased with the size of the attended region, the level of neural activity in a given subregion decreased. These findings are consistent with the physiological predictions of the zoom lens model. Size-related modulations of neural activity were pronounced in early visual areas. We relate this finding to the small receptive field of these areas, whereby only neuronal units with receptive fields covering the attended region received a top-down bias. This preactivation of neuronal units may then have gated selective processing of the features of the object that appeared at the attended location, thus enabling feature integration and object identification. PMID- 12736326 TI - Effects of neurotrophins on synaptic protein expression in the visual cortex of dark-reared rats. AB - Total lack of visual experience [dark rearing (DR)] is known to prolong the critical period and delay development of sensory functions in mammalian visual cortex. Recent results show that neurotrophins (NTs) counteract the effects of DR on functional properties of visual cortical cells and exert a strong control on critical period duration. NTs are known to modulate the development and synaptic efficacy of neurotransmitter systems that are affected by DR. However, it is still unknown whether the actions of NTs in dark-reared animals involve interaction with neurotransmitter systems. We have studied the effects of DR on the expression of key molecules in the glutamatergic and GABAergic systems in control and NT-treated animals. We have found that DR reduced the expression of the NMDA receptor 2A subunit and its associated protein PSD-95 (postsynaptic density-95), of GRIP (AMPA glutamate receptor interacting protein), and of the biosynthetic enzyme GAD (glutamic acid decarboxylase). Returning dark-reared animals to light for 2 hr restored normal expression of the above-mentioned proteins almost completely. NT treatment specifically counteracts DR effects; NGF acts primarily on the NMDA system, whereas BDNF acts primarily on the GABAergic system. Finally, the action of NT4 seems to involve both excitatory and inhibitory systems. These data demonstrate that different NTs counteract DR effects by modulating the expression of key molecules of the excitatory and inhibitory neurotransmitter systems. PMID- 12736327 TI - Sigma 1 receptor-related neuroactive steroids modulate cocaine-induced reward. AB - The sigma1 receptor is critically involved in the rewarding effect of cocaine, as measured using the conditioned place preference (CPP) procedure in mice. Neuroactive steroids exert rapid neuromodulatory effects in the brain by interacting with GABA(A), NMDA, and sigma1 receptors. At the sigma1 receptor level, 3beta-hydroxy-5-androsten-17-one [dehydroepiandrosterone (DHEA)] and 3beta hydroxy-5-pregnen-20-one (pregnenolone) act as agonists, whereas 4-pregnene-3,20 dione (progesterone) is an efficient antagonist. The present study sought to investigate the action of neuroactive steroids in acquisition of cocaine-induced CPP in C57BL/6 mice. None of these steroids induced CPP alone. However, pretreatment with DHEA or pregnenolone (5-20 mg/kg, s.c.) during conditioning with cocaine (10 mg/kg, i.p.) increased the conditioned score. On the contrary, pretreatment with either progesterone (10 or 20 mg/kg, s.c.) or finasteride (25 mg/kg, twice a day), a 5alpha-reductase inhibitor, blocked acquisition of cocaine (20 mg/kg)-induced CPP. A crossed pharmacology was observed between steroids and sigma1 ligands. The sigma1 antagonist N-[2-(3,4-dichlorophenyl)ethyl]-N-methyl-2 (dimethylamino)ethylamine blocked cocaine-induced CPP and its potentiation by DHEA or pregnenolone. Progesterone blocked cocaine-induced CPP and its potentiation by the sigma1 agonist igmesine. These results showed that neuroactive steroids play a role in cocaine-induced appetence, through their interaction with the sigma1 receptor. Therefore, neuroendocrine control of cocaine addiction may not involve solely glucocorticoids. The importance of neuroactive steroids as factors of individual vulnerability to drug addiction should, thus, be considered. PMID- 12736328 TI - The basement membrane components nidogen and type XVIII collagen regulate organization of neuromuscular junctions in Caenorhabditis elegans. AB - Vertebrate neuromuscular junctions (NMJs) contain specialized basal laminas enriched for proteins not found at high concentrations extrasynaptically. Alterations in NMJ basement membrane components can result in loss of NMJ structural integrity and lead to muscular dystrophies. We demonstrate here that the conserved Caenorhabditis elegans basement membrane-associated molecules nidogen/entactin (NID-1) and type XVIII collagen (CLE-1) are associated with axons and particularly enriched near synaptic contacts. NID-1 is concentrated laterally, between the nerve cord and muscles, whereas CLE-1 is concentrated dorsal to the ventral nerve cord and ventral to the dorsal nerve cord, above the regions where synapses form. Mutations in these molecules cause specific and distinct defects in the organization of neuromuscular junctions. The mutant animals exhibit mild movement defects and altered responses to an inhibitor of acetylcholinesterase and a cholinergic agonist, indicating altered synaptic function. Our results provide the first demonstration that basement membrane molecules are important for NMJ formation and/or maintenance in C. elegans and that collagen XVIII and nidogen can have important roles in synapse organization. PMID- 12736329 TI - Functional hemichannels in astrocytes: a novel mechanism of glutamate release. AB - Little is known about the expression and possible functions of unopposed gap junction hemichannels in the brain. Emerging evidence suggests that gap junction hemichannels can act as stand-alone functional channels in astrocytes. With immunocytochemistry, dye uptake, and HPLC measurements, we show that astrocytes in vitro express functional hemichannels that can mediate robust efflux of glutamate and aspartate. Functional hemichannels were confirmed by passage of extracellular lucifer yellow (LY) into astrocytes in nominal divalent cation-free solution (DCFS) and the ability to block this passage with gap junction blocking agents. Glutamate/aspartate release (or LY loading) in DCFS was blocked by multivalent cations (Ca2+, Ba2+, Sr2+, Mg2+, and La3+) and by gap junction blocking agents (carbenoxolone, octanol, heptanol, flufenamic acid, and 18alpha glycyrrhetinic acid) with affinities close to those reported for blockade of gap junction intercellular communication. Glutamate efflux via hemichannels was also accompanied by greatly reduced glutamate uptake. Glutamate release in DCFS, however, was not significantly mediated by reversal of the glutamate transporter: release did not saturate and was not blocked by glutamate transporter blockers. Control experiments in DCFS precluded glutamate release by volume-sensitive anion channels, P2X7 purinergic receptor pores, or general purinergic receptor activation. Blocking intracellular Ca2+ mobilization by BAPTA-AM or thapsigargin did not inhibit glutamate release in DCFS. Divalent cation removal also induced glutamate release from intact CNS white matter (acutely isolated optic nerve) that was blocked by carbenoxolone, suggesting the existence of functional hemichannels in situ. Our results indicated that astrocyte hemichannels could influence CNS levels of extracellular glutamate with implications for normal and pathological brain function. PMID- 12736330 TI - Sp1 and Sp3 are oxidative stress-inducible, antideath transcription factors in cortical neurons. AB - Neuronal cell death in response to oxidative stress may reflect the failure of endogenous adaptive mechanisms. However, the transcriptional activators induced by oxidative stress in neurons that trigger adaptive genetic responses have yet to be fully elucidated. We report that basal DNA binding of the zinc finger transcription factors Sp1 and Sp3 is unexpectedly low in cortical neurons in vitro and is significantly induced by glutathione depletion-induced or hydrogen peroxide-induced oxidative stress in these cells. The increases in Sp1/Sp3 DNA binding reflect, in part, increased levels of Sp1 and Sp3 protein in the nuclei of cortical neurons. Similar induction of Sp1 and Sp3 protein is also observed in neurons in vivo in a chemical or a genetic model of Huntington's disease, two rodent models in which neuronal loss has been attributed to oxidative stress. Sustained high-level expression of full-length Sp1 or full-length Sp3, but not the Sp1 zinc finger DNA-binding domain alone, prevents death in response to oxidative stress, DNA damage, or both. Taken together, these results establish Sp1 and Sp3 as oxidative stress-induced transcription factors in cortical neurons that positively regulate neuronal survival. PMID- 12736331 TI - Single-cell microarray analysis in hippocampus CA1: demonstration and validation of cellular heterogeneity. AB - Laser capture microdissection in combination with microarrays allows for the expression analysis of thousands of genes in selected cells. Here we describe single-cell gene expression profiling of CA1 neurons in the rat hippocampus using a combination of laser capture, T7 RNA amplification, and cDNA microarray analysis. Subsequent cluster analysis of the microarray data identified two different cell types: pyramidal neurons and an interneuron. Cluster analysis also revealed differences among the pyramidal neurons, indicating that even a single cell type in vivo is not a homogeneous population of cells at the gene expression level. Microarray data were confirmed by quantitative RT-PCR and in situ hybridization. We also report on the reproducibility and sensitivity of this combination of methods. Single-cell gene expression profiling offers a powerful tool to tackle the complexity of the mammalian brain. PMID- 12736332 TI - Amiloride-insensitive currents of the acid-sensing ion channel-2a (ASIC2a)/ASIC2b heteromeric sour-taste receptor channel. AB - Acid-sensing ion channel-2a (ASIC2a) is an amiloride-blockable proton-gated cation channel, probably contributing to sour-taste detection in rat taste cells. To isolate another subtype of the sour-taste receptor, we screened a rat circumvallate papilla cDNA library and identified ASIC2b, an N-terminal splice variant of ASIC2a. Reverse transcription-PCR analyses confirmed the expression of ASIC2b transcripts in the circumvallate papilla and, moreover, demonstrated its expression in the foliate and fungiform papillae. Immunohistochemical analyses revealed that ASIC2b, as well as ASIC2a, was expressed in a subpopulation of taste cells in the circumvallate, foliate, and fungiform papillae, and some of the cells displayed both ASIC2a and ASIC2b immunoreactivities. Subsequent coimmunoprecipitation studies with circumvallate papillae extracts indicated that ASIC2b associated with ASIC2a to form assemblies and, together with our immunohistochemical findings, strongly suggested that both ASIC2 subunits formed heteromeric channels in taste cells in the circumvallate, foliate, and fungiform papillae. Oocyte electrophysiology demonstrated that the ASIC2a/ASIC2b channel generated maximal inward currents at a pH of < or =2.0, which is in agreement with the in vivo pH sensitivity of rat taste cells, and that the amiloride sensitivity of the heteromer decreased with decreasing pH and was almost completely abolished at a pH of 2.0. These findings provide persuasive explanations for the amiloride insensitivity of acid-induced responses of rat taste cells. PMID- 12736334 TI - Developmental increase in vesicular glutamate content does not cause saturation of AMPA receptors at the calyx of Held synapse. AB - Whether a quantal packet of transmitter saturates postsynaptic receptors is a fundamental question in central synaptic transmission. However, this question remains open with regard to saturation at mature synapses. The calyx of Held, a giant glutamatergic synapse in the auditory brainstem, becomes functionally mature during the fourth postnatal week in rats. During postnatal development, the mean amplitude of miniature (i.e., quantal) EPSCs (mEPSCs) becomes significantly larger. Experiments using the rapidly dissociating glutamate receptor antagonist kynurenate suggested that vesicular glutamate content increases with development. To test whether AMPA receptors are saturated by a packet of transmitter, we infused a high concentration of l-glutamate into mature calyceal terminals. This caused a marked increase in the mean amplitude of mEPSCs. We conclude that a single packet of transmitter glutamate does not saturate postsynaptic AMPA receptors even at the mature calyx of Held synapse with increased vesicular transmitter content. PMID- 12736333 TI - Scaffolding of Fyn kinase to the NMDA receptor determines brain region sensitivity to ethanol. AB - Alcohol (ethanol) abuse is a major societal problem. Although ethanol is a structurally simple, diffusible molecule, its sites of action are surprisingly selective, and the molecular mechanisms underlying specificity in ethanol actions are not understood. The NMDA receptor channel is one of the main targets for ethanol in the brain. We report here that the brain region-specific compartmentalization of Fyn kinase determines NMDA receptor sensitivity to ethanol. We demonstrate that, in the hippocampus but not in the cerebral cortex, Fyn is targeted to the NR2B subunit of the NMDA receptor by the scaffolding protein RACK1. During acute exposure to ethanol, RACK1 is dissociated from the complex, thereby facilitating Fyn-mediated phosphorylation of NR2B, which enhances channel activity, counteracting the inhibitory actions of ethanol. In this way, the selective scaffolding can account for the ethanol-induced acute tolerance of NMDA receptor activity that is detected in the hippocampus but not in the cerebral cortex. The phosphorylation-dependent, region-specific activities of ethanol on the NMDA receptor provide a compelling molecular explanation that accounts for the selective activities of ethanol and may have important implications for elucidating pathways leading to alcohol addiction. PMID- 12736335 TI - Calcium influx via L- and N-type calcium channels activates a transient large conductance Ca2+-activated K+ current in mouse neocortical pyramidal neurons. AB - Ca2+-activated K+ currents and their Ca2+ sources through high-threshold voltage activated Ca2+ channels were studied using whole-cell patch-clamp recordings from freshly dissociated mouse neocortical pyramidal neurons. In the presence of 4 aminopyridine, depolarizing pulses evoked transient outward currents and several components of sustained currents in a subgroup of cells. The fast transient current and a component of the sustained currents were Ca2+ dependent and sensitive to charybdotoxin and iberiotoxin but not to apamin, suggesting that they were mediated by large-conductance Ca2+-activated K+ (BK) channels. Thus, mouse neocortical neurons contain both inactivating and noninactivating populations of BK channels. Blockade of either L-type Ca2+ channels by nifedipine or N-type Ca2+ channels by omega-conotoxin GVIA reduced the fast transient BK current. These data suggest that the transient BK current is activated by Ca2+ entry through both N- and L-type Ca2+ channels. The physiological role of the fast transient BK current was also examined using current-clamp techniques. Iberiotoxin broadened action potentials (APs), indicating a role of BK current in AP repolarization. Similarly, both the extracellular Ca2+ channel blocker Cd2+ and the intracellular Ca2+ chelator BAPTA blocked the transient component of the outward current and broadened APs in a subgroup of cells. Our results indicate that the outward current in pyramidal mouse neurons is composed of multiple components. A fast transient BK current is activated by Ca2+ entry through high threshold voltage-activated Ca2+ channels (L- and N-type), and together with other voltage-gated K+ currents, this transient BK current plays a role in AP repolarization. PMID- 12736336 TI - Dynamic GABA(A) receptor subtype-specific modulation of the synchrony and duration of thalamic oscillations. AB - Networks of interconnected inhibitory neurons, such as the thalamic reticular nucleus (TRN), often regulate neural oscillations. Thalamic circuits generate sleep spindles and may contribute to some forms of generalized absence epilepsy, yet the exact role of inhibitory connections within the TRN remains controversial. Here, by using mutant mice in which the thalamic effects of the anti-absence drug clonazepam (CZP) are restricted to either relay or reticular nuclei, we show that the enhancement of intra-TRN inhibition is both necessary and sufficient for CZP to suppress evoked oscillations in thalamic slices. Extracellular and intracellular recordings show that CZP specifically suppresses spikes that occur during bursts of synchronous firing, and this suppression grows over the course of an oscillation, ultimately shortening that oscillation. These results not only identify a particular anatomical and molecular target for anti absence drug design, but also elucidate a specific dynamic mechanism by which inhibitory networks control neural oscillations. PMID- 12736337 TI - The role of the hyperpolarization-activated cationic current I(h) in the timing of interictal bursts in the neonatal hippocampus. AB - Under both pathological and experimental conditions, area CA3 of the adult or juvenile hippocampus generates periodic population discharges known as interictal bursts. Whereas the ionic and synaptic basis of individual bursts has been comprehensively studied experimentally and computationally, the pacemaker mechanisms underlying interictal rhythmicity remain conjectural. We showed previously that rhythmic population discharges resembling interictal bursts can be induced in hippocampal slices from first postnatal week mice, in Mg2+-free solution with GABA(A) receptor-mediated inhibition blocked. Here we show that these neonatal bursts occurred with high temporal precision and that their frequency and regularity were greatly reduced by the bradycardic agent ZD-7288 when applied at concentrations and durations that selectively block the hyperpolarization-activated, cationic current I(h). Augmenting I(h) by elevating intracellular cAMP dramatically increased burst frequency in a protein kinase A independent manner. Burst amplitudes were strongly correlated with the preceding, but not the following, interburst intervals. The experimentally observed distribution of interburst intervals was modeled by assuming that a burst was triggered whenever the instantaneous rate of spontaneous EPSPs (sEPSPs) exceeded a threshold and that the mean sEPSP rate was minimal immediately after a burst and then relaxed exponentially to a steady-state level. The effect of blocking I(h) in any given slice could be modeled by decreasing only the steady-state sEPSP rate, suggesting that the instantaneous rate of sEPSPs is governed by the level of I(h) activation and raising the novel possibility that interburst intervals reflected the slow activation kinetics of I(h) in the neonatal CA3. PMID- 12736338 TI - Evidence for long-lasting cholinergic control of gap junctional communication between adrenal chromaffin cells. AB - We investigated long-lasting interactions that may occur between two forms of intercellular signaling: cholinergic synaptic transmission and gap junction mediated coupling in the rat adrenal medulla. The junctional coupling between chromaffin cells was studied during reduced or blocked synaptic transmission in adrenal slices. First, cholinergic synaptic activity was reduced by pharmacological treatment. Bath-application of the nicotinic receptor antagonists hexamethonium, the oxystilbene derivative F3, or alpha-bungarotoxin, acting at distinct neuronal-like postsynaptic nicotinic acetylcholine receptors (nAChRs), significantly increased the incidence of Lucifer yellow passage (dye coupling) between chromaffin cells (p > 0.7 in treated slices vs p = 0.4 in controls). Dye coupling was associated with an elevated macroscopic conductance of the junctional current measured by dual patch-clamp. Pharmacological inhibition of protein trafficking from the trans-Golgi network to the plasma membrane by either brefeldin A or nocodazole pretreatment prevented the effects of nAChR antagonists on dye coupling. Interestingly, this upregulation of gap junction-mediated coupling in response to reduced synaptic activity is of physiological relevance, because it is found in the newborn rat, in which cholinergic synaptic transmission has not yet matured. This mechanism may also be of importance in pathological conditions, because chronic blockade of synaptic transmission after surgical denervation of the adrenal gland also resulted in increased dye coupling between chromaffin cells. In conclusion, our pharmacological, physiological, and pathological data concur to demonstrate that gap junction-mediated intercellular communication between chromaffin cells undergoes persistent adaptation in response to impairment of synaptic activity. These results strongly suggest that gap junctional communication between chromaffin cells is under tonic inhibitory control exerted by cholinergic synaptic inputs. PMID- 12736339 TI - Phosphatidylinositol 3-kinase regulates the induction of long-term potentiation through extracellular signal-related kinase-independent mechanisms. AB - Inhibitors of both phosphatidylinositol-3-kinase (PI3-kinase) and MAPK/ERK (mitogen-activate protein kinase/extracellular signal-related kinase) activation inhibit NMDA receptor-dependent long-term potentiation (LTP). PI3-kinase inhibitors also block activation of ERK by NMDA receptor stimulation, suggesting that PI3-kinase inhibitors block LTP because PI3-kinase is an essential upstream regulator of ERK activation. To examine this hypothesis, we investigated the effects of PI3-kinase inhibitors on ERK activation and LTP induction in the CA1 region of mouse hippocampal slices. Consistent with the notion that ERK activation by NMDA receptor stimulation is PI3-kinase dependent, the PI3-kinase inhibitor wortmannin partially inhibited ERK2 activation induced by bath application of NMDA and strongly suppressed ERK2 activation by high-frequency synaptic stimulation. PI3-kinase and MEK (MAP kinase kinase) inhibitors had very different effects on LTP, however. Both types of inhibitors suppressed LTP induced by theta-frequency trains of synaptic stimulation, but only PI3-kinase inhibitors suppressed the induction of LTP by high-frequency stimulation or low frequency stimulation paired with postsynaptic depolarization. Concentrations of PI3-kinase inhibitors that inhibited LTP when present during high-frequency stimulation had no effect on potentiated synapses when applied after high frequency stimulation, suggesting that PI3-kinase is specifically involved in the induction of LTP. Finally, we found that LTP induced by theta-frequency stimulation was MEK inhibitor insensitive but still PI3-kinase dependent in hippocampal slices from PSD-95 (postsynaptic density-95) mutant mice. Together, our results indicate that the role of PI3-kinase in LTP is not limited to its role as an upstream regulator of MAPK signaling but also includes signaling through ERK-independent pathways that regulate LTP induction. PMID- 12736340 TI - Heme oxygenase-2 protects against lipid peroxidation-mediated cell loss and impaired motor recovery after traumatic brain injury. AB - After traumatic brain injury (TBI), substantial extracellular heme is released from hemoproteins during hemorrhage and cell injury. Heme oxygenase (HO) isozymes are thought to detoxify the pro-oxidant heme to the potent antioxidant, bilirubin. HO-1, the inducible isozyme, is expressed in glial populations after injury and may play a protective role. However, the role of HO-2, the predominant and constitutively expressed isozyme in the brain, remains unclear after TBI. We used a controlled cortical impact injury model to determine the extent and mechanism of damage between HO-2 knock-out (KO) (-/-) and wild-type (WT) (+/+) mice. The specific cellular and temporal expressions of HO-2 and HO-1 were characterized by immunocytochemistry and Western blots. HO-2 was immunolocalized in neurons both before and after TBI, whereas HO-1 was highly upregulated in glia only after TBI. HO activity determined by gas chromatography using brain sonicates from injured HO-2 KO mice was significantly less than that of HO-2 wild types, despite the induction of HO-1 expression after TBI. Cell loss was significantly greater in KO mice in areas including the cortex, the CA3 region of hippocampus, and the lateral dorsal thalamus. Furthermore, motor recovery after injury, as measured by the rotarod assay and an inclined beam-walking task, was compromised in the KO mice. Finally, brain tissue from injured HO-2 KO mice exhibited decreased ability to reduce oxidative stress, as measured with an Fe(2+)/ascorbic acid-mediated carbon monoxide generation assay for lipid peroxidation susceptibility. These findings demonstrate that HO-2 expression protects neurons against TBI by reducing lipid peroxidation via the catabolism of free heme. PMID- 12736341 TI - Learning input correlations through nonlinear temporally asymmetric Hebbian plasticity. AB - Triggered by recent experimental results, temporally asymmetric Hebbian (TAH) plasticity is considered as a candidate model for the biological implementation of competitive synaptic learning, a key concept for the experience-based development of cortical circuitry. However, because of the well known positive feedback instability of correlation-based plasticity, the stability of the resulting learning process has remained a central problem. Plagued by either a runaway of the synaptic efficacies or a greatly reduced sensitivity to input correlations, the learning performance of current models is limited. Here we introduce a novel generalized nonlinear TAH learning rule that allows a balance between stability and sensitivity of learning. Using this rule, we study the capacity of the system to learn patterns of correlations between afferent spike trains. Specifically, we address the question of under which conditions learning induces spontaneous symmetry breaking and leads to inhomogeneous synaptic distributions that capture the structure of the input correlations. To study the efficiency of learning temporal relationships between afferent spike trains through TAH plasticity, we introduce a novel sensitivity measure that quantifies the amount of information about the correlation structure in the input, a learning rule capable of storing in the synaptic weights. We demonstrate that by adjusting the weight dependence of the synaptic changes in TAH plasticity, it is possible to enhance the synaptic representation of temporal input correlations while maintaining the system in a stable learning regime. Indeed, for a given distribution of inputs, the learning efficiency can be optimized. PMID- 12736342 TI - Cytoskeletal and morphological alterations underlying axonal sprouting after localized transection of cortical neuron axons in vitro. AB - We examined the cytoskeletal dynamics that characterize neurite sprouting after axonal injury to cortical neurons maintained in culture for several weeks and compared these with initial neurite development. Cultured neocortical neurons, derived from embryonic day 18 rats, were examined at 3 d in vitro (DIV) and at various time points after axotomy at 21 DIV. The postinjury neuritic response was highly dynamic, progressing through an initial phase of retraction, followed by substantial axonal sprouting within 4-6 hr. Postinjury sprouts were motile and slender with expanded growth cone-like end structures. Microtubule markers were localized to sprout shafts and the proximal regions of putative growth cones and filamentous actin was distributed throughout growth cones, whereas neurofilament proteins were restricted to sprout shafts. A similar distribution of cytoskeletal proteins was present in developing neurites at 3 DIV. Exposure of developing and mature, injured cultures to the microtubule stabilizing agent taxol (10 microg/ml) caused growth inhibition, process distension, the transformation of growth cones into bulbous structures, and abnormal neurite directionality. Microtubule and neurofilament segregation occurred after taxol exposure in developing neurites and postinjury sprouts. Exposure to the microtubule destabilizing agent nocodazole (100 microg/ml) resulted in substantial morphological alteration of developing neurons and inhibited neurite growth and postinjury axonal sprouting. Our results indicate that the axons of cortical neurons have an intrinsic ability to sprout after transection, and similar cytoskeletal dynamics underlie neurite development and postinjury axonal sprouting. PMID- 12736343 TI - Visuomotor behaviors in larval zebrafish after GFP-guided laser ablation of the optic tectum. AB - The optic tectum is the largest visual center in most vertebrates and the main target for retinal ganglion cells (RGCs) conveying visual information from the eye to the brain. The retinotectal projection has served as an important model in many areas of developmental neuroscience. However, knowledge of the function of the tectum is limited. We began to address this issue using laser ablations and subsequent behavioral testing in zebrafish. We used a transgenic zebrafish line that expresses green-fluorescent protein in RGCs projecting to the tectum. By aiming a laser beam at the labeled retinal fibers demarcating the tectal neuropil, the larval tectum could be selectively destroyed. We tested whether tectum-ablated zebrafish larvae, when presented with large-field movements in their surroundings, displayed optokinetic responses (OKR) or optomotor responses (OMR), two distinct visuomotor behaviors that compensate for self-motion. Neither OKR nor OMR were found to be dependent on intact retinotectal connections. Also, visual acuity remained unaffected. Tectum ablation, however, slowed down the OKR by reducing the frequency of saccades but left tracking velocity, gain, and saccade amplitude unaffected. Removal of the tectum had no effect on the processing of second-order motion, to which zebrafish show both OKR and OMR, suggesting that the tectum is not an integral part of the circuit that extracts higher-order cues in the motion pathway. PMID- 12736344 TI - Netrin-1 is a chemorepellent for oligodendrocyte precursor cells in the embryonic spinal cord. AB - Netrin-1, secreted by floor plate cells, orients axon extension in relation to the ventral midline of the embryonic spinal cord. Oligodendrocyte precursor (OP) cells are born close to the ventral midline and migrate away from the floor plate. Here we show that OP cells, identified by expression of the platelet derived growth factor alpha receptor, express the netrin receptors dcc and unc5h1 but do not express netrin-1. Using a microchemotaxis assay, we demonstrate that migrating OPs are repelled by a gradient of netrin-1 in vitro. Furthermore, application of netrin-1 to OPs in vitro triggers retraction of OP processes. In the absence of netrin-1 or Deleted in Colorectal Cancer (DCC) function in vivo, fewer OP cells migrate from the ventral to the dorsal embryonic spinal cord, consistent with netrin-1 acting as a repellent. In addition to their role regulating cell movement, DCC and UNC-5 homologs have been suggested to function as proapoptotic dependence receptors, triggering cell death in the absence of netrin-1. In contrast, we report no evidence of increased OP cell death in vivo or in vitro in the absence of either netrin-1 or DCC. These findings indicate that netrin-1 is a repellent cue for migrating OPs in the embryonic spinal cord. PMID- 12736345 TI - Intracranially administered anti-Abeta antibodies reduce beta-amyloid deposition by mechanisms both independent of and associated with microglial activation. AB - Active immunization against the beta-amyloid peptide (Alphabeta) with vaccines or passive immunization with systemic monoclonal anti-Abeta antibodies reduces amyloid deposition and improves cognition in APP transgenic mice. In this report, intracranial administration of anti-Alphabeta antibodies into frontal cortex and hippocampus of Tg2576 transgenic APP mice is described. The antibody injection resulted initially in a broad distribution of staining for the antibody, which diminished over 7 d. Although no loss of immunostaining for deposited Abeta was apparent at 4 hr, a dramatic reduction in the Alphabeta load was discernible at 24 hr and was maintained at 3 and 7 d. A reduction in the thioflavine-S-positive compact plaque load was delayed until 3 d, at which time microglial activation also became apparent. At 1 week after the injection, microglial activation returned to control levels, whereas Alphabeta and thioflavine-S staining remained reduced. The results from this study suggest a two-phase mechanism of anti Alphabeta antibody action. The first phase occurs between 4 and 24 hr, clears primarily diffuse Alphabeta deposits, and is not associated with observable microglial activation. The second phase occurs between 1 and 3 d, is responsible for clearance of compact amyloid deposits, and is associated with microglial activation. The results are discussed in the context of other studies identifying coincident microglial activation and amyloid removal in APP transgenic animals. PMID- 12736346 TI - Distinct developmental modes and lesion-induced reactions of dendrites of two classes of Drosophila sensory neurons. AB - Little has been understood about the underlying mechanisms that generate the morphological diversity of dendritic trees. Dendritic arborization neurons in Drosophila provide an excellent model system to tackle this question, and they are classified into classes I-IV in order of increasing arbor complexity. Here we have developed transgenic green fluorescent protein markers for class I or class IV cells, which allowed time-lapse recordings of dendritic birth in the embryo, its maturation processes in the larva, and lesion-induced reactions. The two classes used distinct strategies of dendritic emergence from the cell body and branching, which contributed to differences in their basic arbor patterns. In contrast to the class I cells examined, one cell of class IV, which was a focus in this study, continued to elaborate branches throughout larval stages, and it was much more capable of responding to the severing of branches. We also investigated the cellular basis of field formation between adjacent class IV cells. Our results support the fact that class-specific inhibitory interaction is necessary and sufficient for tiling and confirmed that this intercellular communication was at work at individual dendrodendritic interfaces. Finally, this inhibitory signaling appeared to play a central role when arbors of adjacent cells started meeting midway between the cells and until the body wall became partitioned into abutting, minimal-overlapping territories. PMID- 12736347 TI - Long-term depression is not induced by low-frequency stimulation in rat visual cortex in vivo: a possible preventing role of endogenous brain-derived neurotrophic factor. AB - Low-frequency stimulation (LFS) at 1 Hz for 15 min is an effective protocol to induce homosynaptic long-term depression (LTD) in visual cortical slices. It is reported that LFS becomes ineffective when brain-derived neurotrophic factor (BDNF) is applied to slices. It is not known, however, whether such a protocol induces LTD in visual cortex in vivo, and whether endogenous BDNF has the same or similar action. To address these questions, we recorded field potentials of rat visual cortex evoked by stimulation of lateral geniculate nucleus, white matter, or cortical layer IV. We found that LFS did not induce LTD of cortical responses in vivo. To test the possibility that spontaneous activity from retinas would interfere with the induction of LTD, both eyes were removed or inactivated by tetrodotoxin. LTD was not induced in these conditions either. To test whether the difference in temperature between the two preparations is a factor for the discrepancy, the temperature of slices was increased from 31 to 37 degrees C. LTD was induced in slices at either temperature. Then, we hypothesized that endogenous BNDF and its receptors, TrkB, prevent the induction of LTD. To test this, we infused the cortex with an inhibitor of Trk receptor tyrosine kinases, anti-TrkB IgG1, anti-BDNF, and anti-neurotrophin 4/5 antibodies. LTD was induced when the BDNF-TrkB system was blocked. In slices, the level of phosphorylation of Trks was found to decrease with time. These results indicate that activation of TrkB signal pathway prevents LFS from inducing synaptic depression in visual cortex in vivo. PMID- 12736348 TI - A precise retinotopic map of primate striate cortex generated from the representation of angioscotomas. AB - Shadows cast by retinal blood vessels are represented in striate cortex of the squirrel monkey. Their pattern was exploited to generate a true retinotopic map of V1. For calibration, retinal landmarks were projected onto a tangent screen to measure their visual field location. Next, the retina was warped onto striate cortex, distorting it as necessary to match each retinal vessel to its cortical representation. Maps from four hemispheres of two normal adult squirrel monkeys were created and used to derive expressions for cortical magnification factor (M). A mean map was produced by averaging the individual maps. To address the controversial issue of whether the ratio of retinal ganglion cell (RGC) density to M is constant at all eccentricities, we stained a retinal whole mount from one of the two monkeys for Nissl substance. A ganglion cell density map was compiled by sampling the concentration of cells at 171 retinal points. Allowance was made for displaced amacrine cells and for the centripetal displacement of RGCs from central photoreceptors. After these corrections the V1 surface area and RGC density were compared at each eccentricity. The cortical representation of the macula was found to be amplified, even beyond the magnification expected from its high density of RGCs. For example, the central 4 degrees of visual field were allotted 27% of the surface area of V1 but were supplied by only 12% of RGCs. We conclude that, in monkey striate cortex, more tissue is allocated per ganglion cell for the analysis of information emanating from the macula as compared with the peripheral retina. PMID- 12736349 TI - Menstrual cycle-dependent neural plasticity in the adult human brain is hormone, task, and region specific. AB - In rodents, cyclically fluctuating levels of gonadal steroid hormones modulate neural plasticity by altering synaptic transmission and synaptogenesis. Alterations of mood and cognition observed during the menstrual cycle suggest that steroid-related plasticity also occurs in humans. Cycle phase-dependent differences in cognitive performance have almost exclusively been found in tasks probing lateralized neuronal domains, i.e., cognitive domains such as language, which are predominantly executed by one hemisphere. To search for neural correlates of hormonally mediated neural plasticity in humans, we thus conducted a functional magnetic resonance imaging study measuring brain activity related to a semantic decision task in the language domain. This was contrasted with a letter-matching task in the perceptual domain, in which we expected no steroid hormone-mediated effect. We investigated 12 young healthy women in a counterbalanced repeated-measure design during low-steroid menstruation and high steroid midluteal phase. Steroid serum levels correlated with the volume and lateralization of particular brain activations related to the semantic task but not with brain activity related to the perceptual task. More specifically, bilateral superior temporal recruitment correlated positively with progesterone and medial superior frontal recruitment with both progesterone and estradiol serum levels, whereas activations in inferior and middle frontal cortex were unaffected by steroid levels. In contrast to these specific interactions, testosterone levels correlated nonselectively with overall activation levels by neural and/or vascular factor(s). In conclusion, our data demonstrate steroid hormone responsivity in the adult human brain by revealing neural plasticity in the language domain, which appears hormone, task, and region specific. PMID- 12736350 TI - Bandpass filtering at the rod to second-order cell synapse in salamander (Ambystoma tigrinum) retina. AB - The ability to see at night relies on the transduction of single photons by the rod photoreceptors and transmission of the resulting signals through the retina. Using paired patch-clamp recordings, we investigated the properties of the first stage of neural processing of the rod light responses: signal transfer from rods to bipolar and horizontal cells. Bypassing the relatively slow phototransduction process and directly modulating the rod voltage or current allowed us to characterize signal transfer over a wide range of temporal frequencies. We found that the rod to second-order cell synapse acts as a bandpass filter, preferentially transmitting signals with frequencies between 1.5 and 4 Hz while attenuating higher and lower frequency inputs. The similarity of the responses in different types of postsynaptic cell and the properties of miniature EPSCs (mEPSCs) recorded in OFF bipolar cells suggest that most of the bandpass filtering is mediated presynaptically. Modeling of the network of electrically coupled rod photoreceptors suggests that spread of the signal through the network contributed to the observed high-pass filtering but not to the low-pass filtering. Attenuation of low temporal frequencies at the first retinal synapse sharpens the temporal resolution of the light response; attenuation of high temporal frequencies removes voltage noise in the rod that threatens to swamp the light response. PMID- 12736351 TI - Gene microarrays in hippocampal aging: statistical profiling identifies novel processes correlated with cognitive impairment. AB - Gene expression microarrays provide a powerful new tool for studying complex processes such as brain aging. However, inferences from microarray data are often hindered by multiple comparisons, small sample sizes, and uncertain relationships to functional endpoints. Here we sought gene expression correlates of aging dependent cognitive decline, using statistical profiling of gene microarrays in well powered groups of young, mid-aged, and aged rats (n = 10 per group). Animals were trained on two memory tasks, and the hippocampal CA1 region of each was analyzed on an individual microarray (one chip per animal). Aging- and cognition related genes were identified by testing each gene by ANOVA (for aging effects) and then by Pearson's test (correlating expression with memory). Genes identified by this algorithm were associated with several phenomena known to be aging dependent, including inflammation, oxidative stress, altered protein processing, and decreased mitochondrial function, but also with multiple processes not previously linked to functional brain aging. These novel processes included downregulated early response signaling, biosynthesis and activity-regulated synaptogenesis, and upregulated myelin turnover, cholesterol synthesis, lipid and monoamine metabolism, iron utilization, structural reorganization, and intracellular Ca2+ release pathways. Multiple transcriptional regulators and cytokines also were identified. Although most gene expression changes began by mid-life, cognition was not clearly impaired until late life. Collectively, these results suggest a new integrative model of brain aging in which genomic alterations in early adulthood initiate interacting cascades of decreased signaling and synaptic plasticity in neurons, extracellular changes, and increased myelin turnover-fueled inflammation in glia that cumulatively induce aging-related cognitive impairment. PMID- 12736352 TI - Emotion processing in chimeric faces: hemispheric asymmetries in expression and recognition of emotions. AB - Since the discovery of facial asymmetries in emotional expressions of humans and other primates, hypotheses have related the greater left-hemiface intensity to right-hemispheric dominance in emotion processing. However, the difficulty of creating true frontal views of facial expressions in two-dimensional photographs has confounded efforts to better understand the phenomenon. We have recently described a method for obtaining three-dimensional photographs of posed and evoked emotional expressions and used these stimuli to investigate both intensity of expression and accuracy of recognizing emotion in chimeric faces constructed from only left- or right-side composites. The participant population included 38 (19 male, 19 female) African-American, Caucasian, and Asian adults. They were presented with chimeric composites generated from faces of eight actors and eight actresses showing four emotions: happiness, sadness, anger, and fear, each in posed and evoked conditions. We replicated the finding that emotions are expressed more intensely in the left hemiface for all emotions and conditions, with the exception of evoked anger, which was expressed more intensely in the right hemiface. In contrast, the results indicated that emotional expressions are recognized more efficiently in the right hemiface, indicating that the right hemiface expresses emotions more accurately. The double dissociation between the laterality of expression intensity and that of recognition efficiency supports the notion that the two kinds of processes may have distinct neural substrates. Evoked anger is uniquely expressed more intensely and accurately on the side of the face that projects to the viewer's right hemisphere, dominant in emotion recognition. PMID- 12736353 TI - Aberrant chloride transport contributes to anoxic/ischemic white matter injury. AB - Rundown of ionic gradients is a central feature of white matter anoxic injury; however, little is known about the contribution of anions such as Cl-. We used the in vitro rat optic nerve to study the role of aberrant Cl- transport in anoxia/ischemia. After 30 min of anoxia (NaN3, 2 mm), axonal membrane potential (V(m)) decreased to 42 +/- 11% of control and to 73 +/- 11% in the presence of tetrodotoxin (TTX) (1 microm). TTX + 4,4'-diisothiocyanatostilbene-2,2' disulfonic acid disodium salt (500 microm), a broad spectrum anion transport blocker, abolished anoxic depolarization (95 +/- 8%). Inhibition of the K-Cl cotransporter (KCC) (furosemide 100 microm) together with TTX was also more effective than TTX alone (84 +/- 14%). The compound action potential (CAP) area recovered to 26 +/- 6% of control after 1 hr anoxia. KCC blockade (10 microm furosemide) improved outcome (40 +/- 4%), and TTX (100 nm) was even more effective (74 +/- 12%). In contrast, the Cl- channel blocker niflumic acid (50 microm) worsened injury (6 +/- 1%). Coapplication of TTX (100 nm) + furosemide (10 microm) was more effective than either agent alone (91 +/- 9%). Furosemide was also very effective at normalizing the shape of the CAPs. The KCC3a isoform was localized to astrocytes. KCC3 and weaker KCC3a was detected in myelin of larger axons. KCC2 was seen in oligodendrocytes and within axon cylinders. Cl- gradients contribute to resting optic nerve membrane potential, and transporter and channel-mediated Cl- fluxes during anoxia contribute to injury, possibly because of cellular volume changes and disruption of axo-glial integrity, leading to propagation failure and distortion of fiber conduction velocities. PMID- 12736354 TI - Conditional expression in corticothalamic efferents reveals a developmental role for nicotinic acetylcholine receptors in modulation of passive avoidance behavior. AB - Prenatal nicotine exposure has been linked to attention deficit hyperactivity disorder and cognitive impairment, but the sites of action for these effects of nicotine are still under investigation. High-affinity nicotinic acetylcholine receptors (nAChRs) contain the beta2 subunit and modulate passive avoidance (PA) learning in mice. Using an inducible, tetracycline-regulated transgenic system, we generated lines of mice with expression of high-affinity nicotinic receptors restored in specific neuronal populations. One line of mice shows functional beta2 subunit-containing nAChRs localized exclusively in corticothalamic efferents. Functional, presynaptic nAChRs are present in the thalamus of these mice as detected by nicotine-elicited rubidium efflux assays from synaptosomes. Knock-out mice lacking high-affinity nAChRs show elevated baseline PA learning, whereas normal baseline PA behavior is restored in mice with corticothalamic expression of these nAChRs. In contrast, nicotine can enhance PA learning in adult wild-type animals but not in corticothalamic-expressing transgenic mice. When these transgenic mice are treated with doxycycline in adulthood to switch off nAChR expression, baseline PA is maintained even after transgene expression is abolished. These data suggest that high-affinity nAChRs expressed on corticothalamic neurons during development are critical for baseline PA performance and provide a potential neuroanatomical substrate for changes induced by prenatal nicotine exposure leading to long-term behavioral and cognitive deficits. PMID- 12736356 TI - Neural correlates of competing fear behaviors evoked by an innately aversive stimulus. AB - Environment and experience influence defensive behaviors, but the neural circuits mediating such effects are not well understood. We describe a new experimental model in which either flight or freezing reactions can be elicited from mice by innately aversive ultrasound. Flight and freezing are negatively correlated, suggesting a competition between fear motor systems. An unfamiliar environment or a previous aversive event, moreover, can alter the balance between these behaviors. To identify potential circuits controlling this competition, global activity patterns in the whole brain were surveyed in an unbiased manner by c-fos in situ hybridization, using novel experimental and analytical methods. Mice predominantly displaying freezing behavior had preferential neural activity in the lateral septum ventral and several medial and periventricular hypothalamic nuclei, whereas mice predominantly displaying flight had more activity in cortical, amygdalar, and striatal motor areas, the dorsolateral posterior zone of the hypothalamus, and the vertical limb of the diagonal band. These complementary patterns of c-fos induction, taken together with known connections between these structures, suggest ways in which the brain may mediate the balance between these opponent defensive behaviors. PMID- 12736355 TI - Selective enhancement of synaptic inhibition by hypocretin (orexin) in rat vagal motor neurons: implications for autonomic regulation. AB - The hypocretins (orexins) are hypothalamic neuropeptides implicated in feeding, arousal, and autonomic regulation. These studies were designed to determine the actions of hypocretin peptides on synaptic transmission in the dorsal motor nucleus of the vagus nerve (DMV). Whole-cell patch-clamp recordings were made from DMV neurons in transverse slices of rat brainstem. Some of the neurons were identified as gastric-related by retrograde labeling after inoculation of the stomach wall with pseudorabies virus 152, a viral label that reports enhanced green fluorescent protein. Consistent with previous findings, hypocretins caused an inward current (6-68 pA) in most neurons at holding potentials near rest. In addition, the frequency of spontaneous IPSCs was increased in a concentration related manner (up to 477%), with little change in EPSCs. This effect was preserved in the presence of tetrodotoxin, suggesting a presynaptic site of action. Hypocretins increased the amplitude of IPSCs evoked by electrical stimulation of the nucleus tractus solitarius (NTS) but not evoked EPSCs. Hypocretin-induced increases in the frequency of IPSCs evoked by photoactivation of caged glutamate within the NTS were also observed. Identical effects of the peptides were observed in identified gastric-related and unlabeled DMV neurons. In contrast to some previous studies, which have reported primarily excitatory actions of the hypocretins in many regions of the CNS, these data support a role for hypocretin in preferentially enhancing synaptic inhibition, including inhibitory inputs arising from neurons in the NTS. These findings indicate that the hypocretins can modulate and coordinate visceral autonomic output by acting directly on central vagal circuits. PMID- 12736357 TI - Functional dissociation among components of remembering: control, perceived oldness, and content. AB - Remembering is the ability to bring back to mind episodes from one's past and is presumably accomplished by multiple, interdependent processes. In the present functional magnetic resonance imaging study, neural correlates of three hypothesized components of remembering were explored, including those associated with control, perceived oldness, and retrieved content. Levels of each component were separately manipulated by varying study procedures and sorting trials by subject response. Results suggest that specific regions in the left prefrontal cortex, including anterior-ventral Brodmann's Area (BA) 45/47 and more dorsal BA 44, increase activity when high levels of control are required but do not necessarily modulate on the basis of perceived oldness. Parietal and frontal regions, particularly the left parietal cortex near BA 40/39, associate with the perception that information is old and generalize across levels of control and retrieved content. Activity in the parietal cortex correlated with perceived oldness even when judgments were in error. The inferior temporal cortex near BA 19/37 associated differentially with retrieval of visual object content. Within the ventral visual processing stream, content-based modulation was specific to late object-responsive regions, suggesting an efficient retrieval process that spares areas that process more primitive retinotopically mapped visual features. Taken collectively, the results identify neural correlates of distinct components of remembering and provide evidence for a functional dissociation. Frontal regions may contribute to control processes that interact with different posterior regions that contribute a signal that information is old and support the contents of retrieval. PMID- 12736358 TI - Electrophysiological imaging of functional architecture in the cortical middle temporal visual area of Cebus apella monkey. AB - We studied the spatial organization of directionally selective neurons in the cortical middle temporal visual area (area MT) of the Cebus monkey. We recorded neuronal activity from multielectrode arrays as they were stepped through area MT. The set of recording sites in each array penetration described a plane parallel to the cortical layers. At each recording site, we determined the preferred direction of motion. Responses recorded at successive locations from the same electrode in the array revealed gradual changes in preferred direction, along with occasional directional reversals. Comparisons of responses from adjacent electrodes at successive locations enabled electrophysiological imaging of the two-dimensional pattern of preferred directions across the cortex. Our results demonstrate a systematic organization for directionality in area MT of the New World Cebus monkey, which is similar to that known to exist in the Old World macaque. In addition, our results provide electrophysiological confirmation of map features that have been documented in other cortical areas and primate species by optical imaging. Specifically, the tangential organization of directional selectivity is characterized by slow continuous changes in directional preference, as well as lines (fractures) and points (singularities) that fragment continuous regions into patches. These electrophysiological methods also allowed a direct investigation of neuronal selectivities that give rise to map features. In particular, our results suggest that inhibitory mechanisms may be involved in the generation of fractures and singularities. PMID- 12736359 TI - Differential role of mitogen-activated protein kinase in three distinct phases of memory for sensitization in Aplysia. AB - The mitogen-activated protein kinase (MAPK) pathway has been implicated recently in synaptic plasticity and memory. Here we used tail shock-induced sensitization of the tail-elicited siphon withdrawal reflex in Aplysia to examine the role of MAPK in three different phases of memory. We show that a specific pattern of serotonin (5-HT) application that produces intermediate-term and long-term synaptic facilitation (ITF and LTF, respectively) of the sensory-motor (SN-MN) synapses in Aplysia leads to sustained activation of extracellular signal regulated kinase in the ventrocaudal cluster sensory neurons (SNs), which include the tail SNs. Furthermore, repeated tail shocks that induce intermediate-term and long-term memory (ITM and LTM, respectively) for sensitization also lead to sustained MAPK activation in the SNs. Given these results, we next examined the requirement of MAPK activity in (1) SN-MN synaptic facilitation and (2) memory for sensitization in Aplysia, by inhibiting MEK, the upstream kinase that phosphorylates and activates MAPK. In cellular experiments, we show that MAPK activity is required for ITF of tail SN-tail MN synapses, and, in parallel behavioral experiments, we show that ITM requires MAPK activity for its induction but not its expression. In contrast, short-term memory for sensitization does not require MAPK activity. Finally, 5-HT-induced LTF has been shown previously to require MAPK activity. Here we show that LTM for sensitization also requires MAPK activity. These results provide evidence that MAPK plays important roles specifically in long-lasting phases of synaptic plasticity and memory. PMID- 12736360 TI - Estrogen modulates the visceromotor reflex and responses of spinal dorsal horn neurons to colorectal stimulation in the rat. AB - Many gastrointestinal pain syndromes are more prevalent in women than men, suggesting a gonadal steroid influence. We characterized the effects of estrogen on two responses to colorectal distention (CRD) in the rat: the visceromotor reflex (vmr) and L6-S1 dorsal horn neuron activity (ABRUPT and SUSTAINED neurons). Ovariectomized rats were injected with estrogen, and responses to innocuous and noxious intensities of CRD were measured between 4 hr and 14 d after injection and compared with ovariectomized and intact, cycling rats. Plasma estrogen levels were determined at each time point. Ovariectomy significantly decreased the magnitude of the vmr and ABRUPT neuron response to CRD compared with cycling rats. Four and 48 hr after estrogen injection (10 microg), the magnitude of the vmr and ABRUPT neuron response returned to the level or greater than that of cycling rats. All responses were comparable with ovariectomized rats by 7 d. These results paralleled the plasma estrogen concentration. Fifty micrograms of estrogen did not further increase the magnitude of the vmr or neuronal response 48 hr after estrogen but did extend the period of the increased ABRUPT neuron response to 14 d. Estrogen did not affect the response of SUSTAINED neurons. In a separate experiment, the response to innocuous CRD was sensitized in estrogen-treated rats but not ovariectomized or cycling rats. The present data suggest that estrogen modulates the spinal cord processing and reflex responses to innocuous and noxious colorectal stimuli in female rats and may contribute to alterations in sensory processing associated with irritable bowel syndrome. PMID- 12736361 TI - Increased seizure susceptibility and proconvulsant activity of anandamide in mice lacking fatty acid amide hydrolase. AB - A number of recent in vitro studies have described a role for endogenous cannabinoids ("endocannabinoids") as transsynaptic modulators of neuronal activity in the hippocampus and other brain regions. However, the impact that endocannabinoid signals may have on activity-dependent neural events in vivo remains mostly unknown and technically challenging to address because of the short half-life of these chemical messengers in the brain. Mice lacking the enzyme fatty acid amide hydrolase [FAAH (-/-) mice] are severely impaired in their ability to degrade the endocannabinoid anandamide and therefore represent a unique animal model in which to examine the function of this signaling lipid in vivo. Here, we show that the administration of anandamide dramatically augments the severity of chemically induced seizures in FAAH (-/-) mice but not in wild type mice. Anandamide-enhanced seizures in FAAH (-/-) mice resulted in significant neuronal damage in the CA1 and CA3 regions of the hippocampus for the bicuculline and kainate models, respectively. Notably, in the absence of anandamide treatment, FAAH (-/-) mice exhibited enhanced seizure responses to high doses of kainate that correlated with greatly elevated endogenous levels of anandamide in the hippocampus of these animals. Collectively, these studies suggest that both exogenously administered and endogenously produced anandamide display FAAH-regulated proconvulsant activity and do not support a general neuroprotective role for this endocannabinoid in response to excitotoxic stimuli in vivo. More generally, these findings demonstrate that the disinhibitory actions of endocannabinoids observed in hippocampal slices in vitro may also occur in vivo. PMID- 12736362 TI - Brain hyperthermia is induced by methamphetamine and exacerbated by social interaction. AB - Hyperthermia is a symptom of methamphetamine (METH) intoxication and a factor implicated in neurotoxicity during chronic METH use. To characterize the thermic response to METH, it was injected once daily into rats at increasing doses (0, 1, 3, and 9 mg/kg, s.c.) while brain [nucleus accumbens (NAcc), hippocampus] and body (deep temporal muscle) temperatures were continuously monitored. METH produced dose-dependent hyperthermia, with brain structures (especially the NAcc) showing a more rapid and pronounced temperature increase than the muscle. At the highest dose, brain and body temperatures increased 3.5-4.0 degrees C above basal levels and remained elevated for 3-5 hr. Stressful and other high-activity situations such as interaction with a conspecific female are also known to induce a significant hyperthermic response in the rat. A combination of social interaction and METH administration was tested for additive effects. Male rats were exposed daily to a conspecific female for a total of 120 min, and METH was injected at the same doses 30 min after the initial contact with the female. An initial hyperthermic response ( approximately 1.5 degrees C) to social interaction was followed by a large and prolonged hyperthermic response (3.5-5.0 degrees C, 5-7 hr at 9 mg/kg) to METH, which was again stronger in brain structures (especially in the NAcc) than in the muscle. Although the combined effect of the hyperthermic events was not additive, METH administration during social interaction produced stronger and longer-lasting increases in brain and body temperature than that induced by drug alone, heating the brain in some animals near its biological limit (>41 degrees C). PMID- 12736363 TI - Gating of hippocampal-evoked activity in prefrontal cortical neurons by inputs from the mediodorsal thalamus and ventral tegmental area. AB - Projections from the hippocampus, the mediodorsal thalamus (MD), and the ventral tegmental area (VTA) form interconnected neural circuits that converge in the prefrontal cortex (PFC) to participate in the regulation of executive functions. The present study assessed the roles that the MD and VTA play in regulating the hippocampal-PFC pathway using extracellular single-unit recordings in urethane anesthetized rats. MD stimulation inhibited PFC neuron firing (approximately 100 msec duration) evoked by fimbria/fornix (FF) stimulation in a majority of neurons tested. However, this effect was reduced if activation of thalamocortical inputs occurred almost simultaneously (10 msec) with stimulation of the FF. In a separate population of neurons, burst stimulation of the MD produced a short-term (approximately 100 msec) inhibition or facilitation of FF-evoked firing in 66 and 33% of PFC neurons, respectively. Moreover, tetanic stimulation of the MD caused a longer-lasting (approximately 5 min) potentiation of FF-evoked firing. Burst stimulation of the VTA inhibited FF-evoked firing in a frequency-dependent manner: firing evoked by higher-frequency trains of pulses to the FF was less inhibited than firing evoked by single-pulse stimulation. The inhibitory actions of VTA stimulation were augmented by D1 receptor antagonism and attenuated by D2 and D4 antagonists. Moreover, stimulation of the MD 10 msec before stimulation of the FF attenuated the VTA-mediated inhibition of evoked firing. Thus, both the MD and VTA exert a complex gating action over PFC neural activity, either facilitating or inhibiting firing in the hippocampal-PFC pathway depending on the frequency and relative timing of the arrival of afferent input. PMID- 12736365 TI - A within-subjects, within-task demonstration of intact spatial reference memory and impaired spatial working memory in glutamate receptor-A-deficient mice. AB - Gene-targeted mice lacking the AMPA receptor subunit glutamate receptor-A (GluRA) (GluR1) and wild-type controls were compared on a radial-maze task in which the same three of six arms were always baited, but in which the rewards of milk were not replaced within a trial. This procedure allowed not only a within-subjects but also a within-trials assessment of both spatial working memory (WM) and reference memory (RM) in GluRA-/- mice, using identical spatial cues. In experiment 1, the GluRA-/- mice made more WM and RM errors during task acquisition. However, separate groups of GluRA-/- and wild-type mice (experiment 2) acquired a purely RM version of the task at a similar rate, using a paradigm with which it was not possible to make WM errors (doors prevented mice from re entering an arm that they had already visited on that trial). In contrast, mice with hippocampal lesions were dramatically impaired. These results are consistent with the possibility that the WM impairment in the GluRA-/- mice during experiment 1 produced interference that disrupted RM acquisition. A WM component was therefore introduced after RM acquisition in experiment 2 (i.e., the mice were no longer prevented from re-entering a previously visited arm). The GluRA-/- mice now made considerably more WM errors than did wild-type mice, but simultaneously, RM was only mildly and transiently impaired. These experiments provide additional evidence of a selective spatial WM deficit coexisting with intact spatial RM acquisition in GluRA-/- mice, suggesting that different neuronal mechanisms within the hippocampus may support these different kinds of information processing. PMID- 12736364 TI - Salicylate induces tinnitus through activation of cochlear NMDA receptors. AB - Salicylate, the active component of aspirin, is known to induce tinnitus. However, the site and the mechanism of generation of tinnitus induced by salicylate remains unclear. Here, we developed a behavioral procedure to measure tinnitus in rats. The behavioral model was based on an active avoidance paradigm in which rats had to display a motor task (i.e., to jump on a climbing pole when hearing a sound). Giving salicylate led to a decrease in the percentage of correct responses (score) and a drastic increase in the number of false positive responses (i.e., animals execute the motor task during a silent period). Presentation of the sound at a constant perceptive level prevents decrease of the score, leading to the proposal that score is related to hearing performance. In contrast, the increase of false positive responses remained unchanged. In fact, animals behaved as if they hear a sound, indicating that they are experiencing tinnitus. Mefenamate in place of salicylate also increased the number of false positive responses, suggesting that salicylate-induced tinnitus is related to an inhibition of cyclooxygenase. One physiological basis of salicylate ototoxicity is likely to originate from altered arachidonic acid metabolism. Because arachidonic acid potentiates NMDA receptor currents, we tested the involvement of cochlear NMDA receptors in the occurrence of tinnitus. Application of NMDA antagonists into the perilymphatic fluids of the cochlea blocked the increase in pole-jumping behavior induced by salicylate, suggesting that salicylate induces tinnitus through activation of cochlear NMDA receptors. PMID- 12736366 TI - Single amino acid substitution in the mouse IgG1 Fc region induces drastic enhancement of the affinity to protein A. AB - The purification of monoclonal antibody sometimes requires a lot of time and involves complicated steps because of the poorer ability of mouse IgG to interact with protein A, or also with protein G, than IgGs from other species such as those of human and rabbit. To resolve this problem, we exchanged one or two amino acid residues of mouse IgG Fc region with that of human IgG. Three mutants (T252M, T254S and T252M-T254S) showed significant improvement in the affinity to protein A. The exchange of the threonine 252 residue to methionine (T252M) was most efficient. This result suggests that a direct and simple modification allows the efficient purification of monoclonal antibody and of fusion protein containing mouse IgG Fc region. PMID- 12736367 TI - NMR and ICP spectroscopic analysis of the DNA-binding domain of the Drosophila GCM protein reveals a novel Zn2+ -binding motif. AB - Drosophila GCM (glial cell missing) is a novel DNA-binding protein that determines the fate of glial precursors from the neural default to glia. The GCM protein contains the functional domain that is essential for recognition of the upstream sequence of the repo gene. In the DNA-binding region of this GCM protein, there is a cysteine-rich region with which divalent metal ions such as Zn(2+) must bind and other proteins belonging to the GCM family have a corresponding region. To obtain a more detailed insight into the structural and functional features of this DNA-binding region, we have determined the minimal DNA-binding domain and obtained inductively coupled plasma atomic emission spectra and (1)H-(15)N, (1)H-(15)N-(13)C and (113)Cd(2+) NMR spectra, with or without its specific DNA molecule. Considering the results, it was concluded that the minimal DNA-binding domain includes two Zn(2+)-binding sites, one of which is adjacent to the interface for DNA binding. Systematic mutational analyses of the conserved cysteine residues in the minimal DNA-binding domain revealed that one Zn(2+)-binding site is indispensable for stabilization of the higher order structure of this DNA-binding domain, but that the other is not. PMID- 12736369 TI - A protein-protein docking algorithm dependent on the type of complexes. AB - An efficient 'soft docking' algorithm is described to assist the prediction of protein-protein association using three-dimensional structures of molecules. The basic tools are the 'simplified protein' model and the docking algorithm of Wodak and Janin. The side chain flexibility of Arg, Lys, Asp, Glu and Met residues at the protein surface is taken into account. The complex type-dependent filtering technique on the basis of the geometric matching, hydrophobicity and electrostatic complementarity is used to select candidate binding modes. Subsequently, we calculate a scoring function which includes electrostatic and desolvation energy terms. In the 44 complexes tested including enzyme-inhibitor, antibody-antigen and other complexes, native-like structures were all found, of which 30 were ranked in the top 20. Thus, our soft docking algorithm has the potential to predict protein-protein recognition. PMID- 12736368 TI - Designing proteins to crystallize through beta-strand pairing. AB - Inherent difficulties in growing protein crystals are major concerns within structural biology and particularly in structural proteomics. Here, we describe a novel approach of engineering target proteins by surface mutagenesis to increase the odds of crystallizing the molecules. To this end, we have exploited our recent triad-hypothesis using proteins with crystallographically defined beta structures as the principal models. Crystal packing analyses of 182 protein structures belonging to 21 different superfamilies implied that the propensities to crystallize could be engineered into target proteins by replacing short segments, 5-6 residues, of their beta-strands with 'cassettes' of suitable packing motifs. These packing motifs will generate specific crystal packing interactions that promote crystallization. Key features of the primary and tertiary structures of such packing motifs have been identified for immunoglobulins. Further, packing motifs have been engineered successfully into six model antibodies without disturbing their capabilities to be produced, their immunoreactivity and their overall structure. Preliminary crystallization analyses have also been performed. Taken together, the procedures outline a rational protocol for crystallizing proteins by surface mutagenesis. The importance of these findings is discussed in relation to the crystallization of proteins in general. PMID- 12736370 TI - Functional sites and evolutionary connections of acylhomoserine lactone synthases. AB - Acylhomoserine lactone (AHL) synthases act as chemical communication signals or pheromones in Gram-negative bacteria and regulate diverse physiological events in a cell density-dependent manner. The recent crystal structure determination of EsaI, a key enzyme in this pathway, shows that the AHL synthase superfamily members adopt the fold of the N-acetyltransferase superfamily. We suggest, by the identification of intermediate sequences, that the two superfamilies are evolutionarily related. Evolutionary trace analyses of aligned sequences and docking studies have been used to discuss functionally important residues of EsaI homologues. PMID- 12736371 TI - A simple electrostatic criterion for predicting the thermal stability of proteins. AB - The enhancement of protein thermostability is an important issue for both basic science and biotechnology purposes. We have developed a thermostability criterion for a protein in terms of a quasi-electric dipole moment (contributed by its charged residues) defined for an electric charge distribution whose total charge is not zero. It was found that minimization of the modulus of this dipole moment increased its thermal stability, as demonstrated by surveying these values in pairs of mesostable-thermostable homologous proteins and in mutations described in the literature. The analysis of these observations provides criteria for thermostabilization of a protein, by computing its dipole profile. This profile is obtained by direct substitution of each amino acid of the sequence by either a positive, negative or neutral amino acid, followed by a recalculation of the dipole moment. As an experimental example, these criteria were applied to a beta glucanase to enhance its thermal stability. PMID- 12736372 TI - Hyperthermostabilization of Bacillus licheniformis alpha-amylase and modulation of its stability over a 50 degrees C temperature range. AB - Bacillus licheniformis alpha-amylase (BLA) is a highly thermostable starch degrading enzyme that has been extensively studied in both academic and industrial laboratories. For over a decade, we have investigated BLA thermal properties and identified amino acid substitutions that significantly increase or decrease the thermostability. This paper describes the cumulative effect of some of the most beneficial point mutations identified in BLA. Remarkably, the Q264S N265Y double mutation led to a rather limited gain in stability but significantly improved the amylolytic function. The most hyperthermostable variants combined seven amino acid substitutions and inactivated over 100 times more slowly and at temperatures up to 23 degrees C higher than the wild-type enzyme. In addition, two highly destabilizing mutations were introduced in the metal binding site and resulted in a decrease of 25 degrees C in the half-inactivation temperature of the double mutant enzyme compared with wild-type. These mutational effects were analysed by protein modelling based on the recently determined crystal structure of a hyperthermostable BLA variant. Our engineering work on BLA shows that the thermostability of an already naturally highly thermostable enzyme can be substantially improved and modulated over a temperature range of 50 degrees C through a few point mutations. PMID- 12736373 TI - Solubility engineering of the HhaI methyltransferase. AB - DNA methylation is involved in epigenetic control of numerous cellular processes in eukaryotes, however, many mechanistic aspects of this phenomenon are not yet understood. A bacterial prototype cytosine-C5 methyltransferase, M.HhaI, serves as a paradigm system for structural and mechanistic studies of biological DNA methylation, but further analysis of the 37 kDa protein is hampered by its insufficient solubility (0.15 mM). To overcome this problem, three hydrophobic patches on the surface of M.HhaI that are not involved in substrate interactions were subjected to site-specific mutagenesis. Residues M51 or V213 were substituted by polar amino acids of a similar size, and/or the C-terminal tetrapeptide FKPY was replaced by a single glycine residue (Delta324G). Two out of six mutants, delta324G and V213S/delta324G, showed improved solubility in initial analyses and were purified to homogeneity using a newly developed procedure. Biochemical studies of the engineered methyltransferases showed that the deletion mutant delta324G retained identical DNA binding, base flipping and catalytic properties as the wild-type enzyme. In contrast, the engineered enzyme showed (i) a significantly increased solubility (>0.35 mM), (ii) high-quality 2D [(15)N,(1)H] TROSY NMR spectra, and (iii) (15)N spin relaxation times evidencing the presence of a monomeric well-folded protein in solution. PMID- 12736374 TI - Stabilization and activation of alpha-chymotrypsin in water-organic solvent systems by complex formation with oligoamines. AB - Formation of enzyme-oligoamine complexes was suggested as an approach to obtain biocatalysts with enhanced resistance towards inactivation in water-organic media. Complex formation results in broadening (by 20-40% v/v ethanol) of the range of cosolvent concentrations where the enzyme retains its catalytic activity (stabilization effect). At moderate cosolvent concentrations (20-40% v/v) complex formation activates the enzyme (by 3-6 times). The magnitude of activation and stabilization effects increases with the number of possible electrostatic contacts between the protein surface and the molecules of oligoamines (OA). Circular dichroism spectra in the far-UV region show that complex formation stabilizes protein conformation and prevents aggregation in water-organic solvent mixtures. Two populations of the complexes with different thermodynamic stabilities were found in alpha-chymotrypsin (CT)-OA systems depending on the CT/OA ratio. The average dissociation constants and stoichiometries of both low- and high-affinity populations of the complexes were estimated. It appears that it is the low-affinity sites on the CT surface that are responsible for the activation effect. PMID- 12736375 TI - The LLSGIV stretch of the N-terminal region of HIV-1 gp41 is critical for binding to a model peptide, T20. AB - A number of peptides and peptide analogs derived from the membrane proximal region of gp41 ectodomain are found to be effective inhibitors of human immunodeficiency virus type 1 (HIV-1)-mediated fusion events. One of them, T20 (aa 638-673), was found disordered and sparingly soluble in water, but became soluble upon mixing with selected, structured peptides from the amino terminal heptad repeat (HR1) region of gp41 using a simple and sensitive method of reduction in the scattering of T20 suspension. From the results on mapping the locus of interaction with T20 by employing partially overlapping peptides derived from HR1, it was concluded that the LLSGIV segment was a critical docking site for the C-terminal peptide of gp41 in its putative inhibitory action consistent with a previous fluorescence study. It was also found that peptides capable of solubilizing T20 dispersion have a high content of helix, as well as beta-strand, conformation in aqueous solution. Specificity of T20/HR1-derived peptide binding was ascertained by using a scrambled sequence of a T20-active peptide and a plateau in scattering reduction of T20 suspension with variation in the concentration of a T20-active HR1 peptide. Implications on the mechanism of T20 inhibition and the sequence of folding of the gp41 core structure are discussed. PMID- 12736376 TI - Differential expression of growth factors at the cellular level in virus-infected brain. AB - The contribution of host factors to rabies virus (RV) transcription/replication and axonal/transsynaptic spread is largely unknown. We previously identified several host genes that are up-regulated in the mouse brain during RV infection, including neuroleukin, which is involved in neuronal growth and survival, cell motility, and differentiation, and fibroblast growth factor homologous factor 4 (FHF4), which has been implicated in limb and nervous system development. In this study, we used real-time quantitative RT-PCR to assess the expression of mRNAs specific for neuroleukin, the two isoforms of FHF4 (FHF4-1a and -1b) encoded by the FHF4 gene, and N protein of RV in neurons and astrocytes isolated by laser capture microdissection from mouse brains infected with the laboratory-adapted RV strain CVS-N2c or with a street RV of silver-haired bat origin. Differences in the gene expression patterns suggest that the capacity of RV strains to infect nonneuronal cells and differentially modulate host gene expression may be important in virus replication and spread in the CNS. PMID- 12736377 TI - Adoptive transfer of apoptotic splenocytes worsens survival, whereas adoptive transfer of necrotic splenocytes improves survival in sepsis. AB - In sepsis, both necrotic and apoptotic cell death can occur. Apoptotic cells induce anergy that could impair the host response, whereas necrotic cells cause immune activation that might result in enhanced antimicrobial defenses. We determined whether adoptive transfer of apoptotic or necrotic cells impacted survival in a clinically relevant sepsis model. We also evaluated the effects of adoptive transfer of apoptotic or necrotic cells on the prototypical TH1 and TH2 cytokines IFN-gamma and IL-4, respectively. C57BL6/J mice had adoptive transfer of apoptotic (irradiated) or necrotic (freeze thaw) splenocytes. Controls received saline. Apoptotic cells greatly increased mortality, whereas necrotic splenocytes markedly improved survival, P < or = 0.05. The contrasting effects that apoptotic or necrotic cells exerted on survival were mirrored by opposite effects on splenocyte IFN-gamma production with greatly decreased and increased production, respectively. Importantly, either administration of anti-IFN-gamma antibodies or use of IFN-gamma knockout mice prevented the survival benefit occurring with necrotic cells. This study demonstrates that the type of cell death impacts survival in a clinically relevant model and identifies a mechanism for the immune suppression that is a hallmark of sepsis. Necrotic cells (and likely apoptotic cells) exert their effects via modulation of IFN-gamma PMID- 12736378 TI - Picosecond conformational transition and equilibration of a cyclic peptide. AB - Ultrafast IR spectroscopy is used to monitor the nonequilibrium backbone dynamics of a cyclic peptide in the amide I vibrational range with picosecond time resolution. A conformational change is induced by means of a photoswitch integrated into the peptide backbone. Although the main conformational change of the backbone is completed after only 20 ps, the subsequent equilibration in the new region of conformational space continues for times >16 ns. Relaxation and equilibration processes of the peptide backbone occur on a discrete hierarchy of time scales. Albeit possessing only a few conformational degrees of freedom compared with a protein, the peptide behaves highly nontrivially and provides insights into the complexity of fast protein folding. PMID- 12736379 TI - Two Arabidopsis circadian oscillators can be distinguished by differential temperature sensitivity. AB - Circadian rhythms are widespread in nature and reflect the activity of an endogenous biological clock. In metazoans, the circadian system includes a central circadian clock in the brain as well as distinct clocks in peripheral tissues such as the retina or liver. Similarly, plants have distinct clocks in different cell layers and tissues. Here, we show that two different circadian clocks, distinguishable by their sensitivity to environmental temperature signals, regulate the transcription of genes that are expressed in the Arabidopsis thaliana cotyledon. One oscillator, which regulates CAB2 expression, responds preferentially to light-dark versus temperature cycles and fails to respond to the temperature step associated with release from stratification. The second oscillator, which regulates CAT3 expression, responds preferentially to temperature versus light-dark cycles and entrains to the release from stratification. Finally, the phase response curves of these two oscillators to cold pulses are distinct. The phase response curve of the oscillator component TOC1 to cold pulses is similar to that of CAB2, indicating that CAB2 is regulated by a TOC1-containing clock. The existence of two clocks, distinguishable on the basis of their sensitivity to temperature, provides an additional means by which plants may integrate both photoperiodic and temperature signals to respond to the changing seasons. PMID- 12736381 TI - Development of microbial-human enterocyte interaction: cholera toxin. AB - Diarrhea in infants and children is a major health hazard worldwide. Certain toxigenic diarrheas occur more commonly and are manifested more severely during the neonatal period. We have previously studied the regulation of cholera toxin induced secretion in animal models during development. In those studies we have shown that cholera toxin stimulates a much greater secretion by immature compared with mature small intestine, and the mechanism appears to be an up-regulation of postreceptor signal transduction molecules (adenyl cyclase and Gsalpha) leading to an elevated cAMP level. In this study, using experimental models of human intestinal development (fetal cell lines, a micro-Ussing chamber, organ cultures, and fetal intestinal xenograft transplants), we provide preliminary evidence that cholera toxin induces an enhanced secretion mediated in part by a developmental up-regulation of the cAMP response in immature versus mature human small intestine. Additional studies are needed, however, to further define whether other developmental events (e.g. receptor expression) also regulate cholera toxin enterocyte-enhanced interaction. Nonetheless, this approach to determining the role of development in the pathophysiology of cholera in infants may help in strategies to prevent and treat this condition and other age-related intestinal infectious diseases. PMID- 12736380 TI - Endogenous surfactant metabolism in newborn infants with and without respiratory failure. AB - Studies using stable isotopically labeled glucose and palmitate as precursors of pulmonary surfactant synthesis have demonstrated slow surfactant turnover in premature infants with respiratory distress syndrome (RDS). However, only limited data about surfactant turnover are available for term infants. Because acetate is a direct precursor of de novo synthesized surfactant fatty acid, we measured [1 13C1]acetate incorporation into surfactant of term infants without respiratory dysfunction (control group), preterm infants with RDS, and term infants with primary respiratory failure to determine whether stable isotopically labeled acetate would yield similar results to previous studies of preterm infants with RDS and, furthermore, would distinguish normal from abnormal surfactant turnover. Despite similar amounts of phospholipids and acetate precursor enrichment, the control group had higher fractional synthetic rate and shorter half-life of clearance than preterm infants with RDS, (fractional synthetic rate, 15.4 +/- 2.4 versus 2.2 +/- 0.4%/d, p < 0.001; half-life of clearance, 27 +/- 3 versus 105 +/- 11 h, p < 0.001). Term infants with severe respiratory failure had a lower fractional synthetic rate than those with mild disease (2.9 +/- 0.6 versus 13.8 +/- 3.5%/d, p = 0.014) and a reduced amount of phospholipids recovered from tracheal aspirates (54 +/- 17 versus 300 +/- 28 nmol, severe versus mild disease, respectively, p < 0.001). The amount of phospholipids in tracheal aspirates correlated inversely with disease severity, (r = -0.75, p = 0.01). We conclude that normal surfactant turnover in term infants is faster than in preterm infants with RDS. Surfactant turnover in term infants with severe respiratory failure is similar to that of preterm infants with RDS, suggesting either delayed maturity of the surfactant system or disruption from the underlying disease process. PMID- 12736383 TI - Rare disorders of metabolism with elevated butyryl- and isobutyryl-carnitine detected by tandem mass spectrometry newborn screening. AB - Tandem mass spectrometry was adopted for newborn screening by North Carolina in April 1999. Since then, three infants with short-chain acyl-CoA dehydrogenase (SCAD) and one with isobutyryl-CoA dehydrogenase deficiency were detected on the basis of elevated butyrylcarnitine/isobutyrylcarnitine (C4-carnitine) concentrations in newborn blood spots analyzed by tandem mass spectrometry. For three SCAD-deficient infants, biochemical evaluation included a plasma acylcarnitine profile with markedly elevated C4-carnitine, urine organic acid analysis with markedly elevated ethylmalonic and 2-methylsuccinic acids, and markedly elevated [U-13C]butyrylcarnitine concentrations in medium from fibroblasts incubated with [U-13C]palmitic acid and excess l-carnitine, consistent with classic SCAD deficiency. Two of three infants diagnosed with classic SCAD deficiency remained asymptomatic; however, the third infant presented with seizures and a cerebral infarct at 10 wk of age. All three infants had putatively inactivating mutations in both alleles of the SCAD gene. The highly elevated plasma C4-carnitine levels in the three infants detected by newborn screening tandem mass spectrometry differentiated them from infants and children who were homozygous or compound heterozygous for one of two SCAD gene susceptibility variations; for the latter group the C4-carnitine levels were normal. Isobutyryl-CoA dehydrogenase deficiency in a fourth infant was confirmed after isolated elevation of C4-carnitine in the acylcarnitine profile. PMID- 12736382 TI - Morphine enhances HIV infection of neonatal macrophages. AB - Perinatal transmission of HIV accounts for almost all new HIV infections in children. There is an increased risk of perinatal transmission of HIV with maternal illicit substance abuse. Little is known about neonatal immune system alteration and subsequent susceptibility to HIV infection after morphine exposure. We investigated the effects of morphine on HIV infection of neonatal monocyte-derived macrophages (MDM). Morphine significantly enhanced HIV infection of neonatal MDM. Morphine-induced HIV replication in neonatal MDM was completely suppressed by naltrexone, the opioid receptor antagonist. Morphine significantly up-regulated CCR5 receptor expression and inhibited the endogenous production of macrophage inflammatory protein-1beta in neonatal MDM. Thus, morphine, most likely through alteration of beta-chemokines and CCR5 receptor expression, enhances the susceptibility of neonatal MDM to HIV infection, and may have a cofactor role in perinatal HIV transmission and infection. PMID- 12736385 TI - Predictive value of brain-specific proteins in serum for neurodevelopmental outcome after birth asphyxia. AB - Brain-specific proteins have been used to detect cerebral injury after birth asphyxia. Previous investigations suggest that serum protein S-100beta, brain specific creatine kinase (CK-BB), and neuron-specific enolase (NSE) are capable of identifying patients with a risk of developing hypoxic-ischemic encephalopathy. Whether detection of elevated serum concentrations of these proteins reflects long-term neurodevelopmental impairment remains to be investigated. We examined serum protein S-100beta, NSE, and CK-BB at 2, 6, 12, and 24 h after birth in 29 asphyxiated infants and 20 control infants. Neurodevelopmental follow-up examinations were performed at 20 mo of age using the German revision of the Griffiths scales for developmental assessment. Elevated concentrations of serum protein S-100beta, NSE, and CK-BB within 24 h after asphyxia did not correlate with long-term neurodevelopmental delay. We conclude that serum protein S-100beta, NSE, and CK-BB, sampled on the first day of life, is of limited value in predicting severe brain damage after birth asphyxia. PMID- 12736384 TI - Effects of hypoxia, hypoglycemia, and muscle shortening on cell death in the sheep ductus arteriosus. AB - After birth, constriction of the full-term ductus arteriosus produces ischemic hypoxia, caspase activation, DNA fragmentation (>70% of cell nuclei are positive by the terminal deoxynucleotidyl transferase nick-end labeling [TUNEL] technique), and permanent ductus closure. In contrast, the preterm ductus frequently fails to develop these changes. We used the TUNEL technique to examine rings of fetal ductus arteriosus (incubated for 24 h at different oxygen and glucose concentrations) to determine the roles of 1) constriction and shortening, 2) hypoxia, and 3) hypoglycemia in producing cell death. Under controlled conditions, late-gestation ductus rings had a low rate of TUNEL-positive staining (0.6 +/- 0.9%) that did not change during muscle shortening. Although hypoxia (6.9 +/- 3.5%) and hypoglycemia (2.4 +/- 1.9%) increased the incidence of TUNEL positive staining, only the combination of hypoxia-plus-hypoglycemia increased the incidence to the range found in vivo (83 +/- 9.5%). The combination of hypoxia-plus-hypoglycemia was associated with an oligonucleosomal pattern of DNA fragmentation. Under the same experimental conditions, the preterm ductus was capable of developing a similar degree of TUNEL-positive staining as found at term. Although caspase-3 and caspase-7 were activated in rings exposed to hypoxia plus-hypoglycemia, a nonselective caspase inhibitor, Z-VAD.FMK (which inhibited caspase-3 and caspase-7 cleavage in the rings), did not diminish the degree of TUNEL-positive staining. We hypothesize that the preterm ductus is capable of developing an extensive degree of cell death, if it can develop the same degree of hypoxia and hypoglycemia found in the full-term newborn ductus. We also hypothesize that cell death in the ductus wall may involve pathways that are not dependent on caspase-3 or -7 activation. PMID- 12736386 TI - Selective cortical alteration after hypoxic-ischemic injury in the very immature rat brain. AB - Distinctive cerebral lesions with disruptions to the developing white matter are found in very low birth weight (VLBW) infants. Although hypoxia-ischemia (HI) is a causal pathway, the pathogenesis of cerebral white matter injury in the VLBW infant is not fully understood. Pertinent murine models would facilitate the investigation of the processes leading to these cerebral lesions and enable the evaluation of therapeutic strategies. Postnatal d 3 (P3) rats are at a stage of cortical oligodendroglial maturation and axonal outgrowth similar to very preterm infants. Our aim was to characterize the effects of a focal hypoxic-ischemic injury at P3 on subsequent cerebral development. Three groups of P3 Wistar rats were investigated: group I underwent right carotid ligation followed by 6% hypoxia for 30 min (HI), group 2 had carotid ligation only, and group 3 had no intervention. At P21, in the HI group, the right cortical area was reduced compared with controls (p < 0.01). There were no significant alterations in the size of the dorsal hippocampus, striatum, and thalamus. The cortical myelinated area was reduced in the HI animals compared with controls (p < 0.01). There was a corresponding loss of myelinated axons extending up into the cortex, with deep cortical neuronal and axonal architecture markedly disrupted. Glial fibrillary acidic protein immunohistology showed a reactive gliosis in the deep parietal cortex (p < 0.01). Moderate HI injury in the immature rat brain compromised cortical growth and led to a selective alteration of cortical myelinated axons with persistent gliosis. These alterations induced at P3 by unilateral HI share neuropathological similarities with the diffuse white matter lesions found in VLBW infants. PMID- 12736387 TI - Inherited mitochondrial DNA depletion. AB - Mitochondrial DNA (mtDNA) depletion is associated with heterogeneous clinical phenotypes. The recent identification of the mutated genes in three groups of patients with mtDNA depletion had underscored the importance of the synthetic pathway of the mitochondrial nucleotides for mtDNA replication. Future goals include understanding how the defective proteins perturb replication, why it affects only some tissues and spares others, and which other genes should be considered in other patients with mtDNA depletion. PMID- 12736388 TI - The influence of prematurity and long chain polyunsaturate supplementation in 4 week adjusted age baboon neonate brain and related tissues. AB - Clinical studies show that docosahexaenoic acid (DHA) and arachidonic acid (ARA) supplemented formula improve visual function in preterm infants, however improved fatty acid status is known only for plasma and red blood cells (RBC) since target organs cannot be sampled from humans. Baboons were randomized to one of four groups: Term breast-fed (B); Term formula-fed (T-); Preterm formula-fed (P-); and Preterm DHA/ARA-supplemented formula-fed (P+). The P+ contained 0.61 +/- 0.03% DHA and 1.21 +/- 0.09% ARA, and breast milk had 0.68 +/- 0.22% and 0.62 +/- 0.12% as DHA and ARA, respectively. The B and P+ groups had significantly higher DHA concentration in all tissues than T- and P-. The P- group showed dramatically lower DHA content of 35%, 27%, 66%, and 75% in the brain, retina, liver, and plasma, respectively, compared with B. Supplementation prevented declines in DHA levels in the retina, and liver, and attenuated the decline in brain, plasma and RBC of preterm animals. In contrast, ARA was not significantly lower compared with B in any group in any tissue but was significantly elevated in liver and brain. RBC and plasma DHA were correlated with DHA in tissues; RBC/plasma ARA were uncorrelated with tissue ARA. We conclude that 1) DHA drops precipitously in term and preterm primates consuming formula without long chain polyunsaturates, while 22:5n-6 concentration rises; 2) tissue ARA levels are insensitive to dietary LCP supplementation or prematurity, 3) plasma and RBC levels of ARA are uncorrelated with total ARA levels; 4) DHA levels are correlated with group effects and are uncorrelated within groups. PMID- 12736389 TI - Meconium induces only localized inflammatory lung injury in piglets. AB - Neonatal meconium aspiration often produces severe respiratory distress due to an inflammatory pulmonary injury, but the extension of this damaging reaction to the noncontaminated lung regions is still uncertain. To investigate the presence of generalized pulmonary inflammatory response, 31 anesthetized and ventilated neonatal piglets (1-3 d) were studied. Meconium (n = 16) or saline (n = 15) was instilled unilaterally into the right lung, and analysis of the lung tissue or bronchoalveolar lavage (BAL) fluid from both lungs was performed after 12 h. Meconium increased the wet/dry weight ratio, histologic tissue injury score and tissue myeloperoxidase activity as well as BAL fluid total cell count in the contaminated lung. Tumor necrosis factor-alfa concentrations in BAL fluid did not however differ significantly. Furthermore, in the meconium-instilled lungs the tissue and lavage fluid catalytic activity of phospholipase A2 (PLA2) and tissue PLA2 group-I and group-II concentrations were significantly elevated. Although BAL fluid catalytic activity of PLA2 was moderately increased also in the meconium noninstilled lung, significant inflammatory injury in this lung was absent. The results thus indicate that meconium aspiration induces severe local inflammation and lung injury, but significant generalized pulmonary inflammatory damage in the pathogenesis of meconium aspiration syndrome is unlikely. PMID- 12736390 TI - Hypothermia for 24 hours after asphyxic cardiac arrest in piglets provides striatal neuroprotection that is sustained 10 days after rewarming. AB - The neuroprotective effect of hypothermia instituted after resuscitation from asphyxic cardiac arrest has not been studied in immature brain, particularly in a large animal model with recovery periods greater than 4 d. Moreover, protection from severe hypoxia seen with 3 h of hypothermia was reported to be lost when hypothermic duration was extended to 24 h in unsedated piglets, in contrast to the neuroprotection reported by 72 h of intrauterine head cooling in fetal sheep. Piglets (5-7 postnatal days) were subjected to asphyxic cardiac arrest followed by 24 h of either hypothermia (34 degrees C) or normothermia (38.5-39 degrees C). Comparisons were made with normothermic and hypothermic surgical sham animals without asphyxia. All of these groups were sedated, paralyzed, and mechanically ventilated for the first 24 h to prevent shivering and possible depletion of glucose stores. Hypothermia per se did not cause remarkable structural abnormalities. Ischemic damage was evaluated in putamen at 1 d of recovery without rewarming and at 11 d (10 d +/- SD after rewarming). Ischemic cytopathology affected 60 +/- 12% of neurons in putamen of normothermic animals compared with 9 +/- 6% in hypothermic animals at 1 d of recovery without rewarming. At 11 d of recovery from hypoxia-ischemia, the density of viable neurons (neuron profiles/mm2) in putamen was markedly reduced in normothermic animals (81 +/- 40) compared with hypothermic animals (287 +/- 22), which was the same as in sham normothermic (271 +/- 21), sham hypothermic (288 +/- 46) and naive animals (307 +/- 51). These data demonstrate that 24 h of hypothermia at 34 degrees C with sedation and muscle relaxation after asphyxic cardiac arrest prevents necrotic striatal neuronal cell death in immature brain before rewarming, and that the effect is sustained at 11 d after injury without deleterious side effects. PMID- 12736391 TI - Knee articular cartilage development in children: a longitudinal study of the effect of sex, growth, body composition, and physical activity. AB - The aim of this study was to describe the effect of sex, growth, Tanner stage, and physical activity on knee articular cartilage volume development. A total of 74 randomly selected male and female children aged 9-18 y were measured on two occasions at an average interval of 1.6 y (range 1.3-1.9). Articular cartilage volume was determined at the patella, medial tibial, and lateral tibial compartments by processing images acquired in the sagittal plane using T1 weighted fat saturation magnetic resonance. Height, weight, and BMI were measured while Tanner stage and physical activity were assessed by questionnaire. Articular cartilage volume increased at all sites peaking in Tanner stage two. Males gained articular cartilage faster than females at all sites (patella +233 microL/y, 95% CI -7, +473, medial tibial +350 microL/y, 95% CI +118, +582, lateral tibial +256 microL/y, 95% CI +22, +488). In both sexes, articular cartilage volume accrual at tibial but not patella sites correlated significantly with height change but not weight change. Overweight children did not differ significantly from normal children in articular cartilage volume either cross sectionally or longitudinally. The most consistent physical activity association was with average intensity of sport with those above the median gaining approximately twice as much as those below the median at tibial (p < 0.05) but not patella sites. In conclusion, most children gain articular cartilage during growth, but there is wide variation in the amount of articular cartilage accrual. In particular, younger children, males, and those undertaking more vigorous sports have substantially higher accrual rates. These results provide novel data on articular cartilage development in humans. The long-term significance of these results with regard to osteoarthritis of the knee in later life remains hypothetical. PMID- 12736392 TI - Gestational pattern of heme oxygenase expression in the rat. AB - Fetal growth is influenced by many intrinsic and extrinsic factors. Our objective was to determine the pattern of heme oxygenase (HO) expression in the pregnant rat and to study its association with fetal growth and growth factors. Uterine tissues were obtained from nonpregnant and from time-mated rats at 7, 13, 16, 19, and 21 d of pregnancy. Placental tissue was obtained on d 13, 16, 19 and 21 of pregnancy. Tissues were evaluated for HO activity, HO-1, HO-2, leptin and vascular endothelial growth factor protein, and HO-1 and HO-2 mRNA. HO activity in both the uterus and placenta peaked on d 21 of pregnancy. In the uterus, HO-1 and HO-2 protein and total mRNA levels peaked on d 16 of pregnancy, whereas, in the placenta, HO-1 and HO-2 protein levels peaked on d 19. Additionally, placental HO-1 mRNA peaked on d 16, but placental HO-2 mRNA declined toward the end of pregnancy. Placental leptin and vascular endothelial growth factor protein levels followed a similar pattern to placental HO-1 and peaked on d 16. We conclude that there is a clear uterine and placental gestational pattern of HO expression in the rat. This pattern is comparable to that of vascular endothelial growth factor and leptin. PMID- 12736394 TI - Sleep disturbances in children with attention deficit hyperactivity disorder. AB - The objective of the study was to compare polysomnographic patterns in two groups of preadolescent children with attention deficit hyperactivity disorder (ADHD) (a sleep clinic referral sample and a community sample) with control children. A prospective and observational study in the sleep clinic and the community was undertaken. Forty-seven ADHD children referred to the sleep clinic (ADHDcl), 53 ADHD children from a community survey (ADHDcom), and 49 control children underwent overnight polysomnography. Significant differences between the groups were observed for rapid-eye-movement (REM) sleep latency and percentage, and periodic limb movement index with associated arousals (PLMa). REM sleep latency was shorter in controls than ADHDcl and ADHDcom (p < 0.01) and REM% was highest in controls and lowest in ADHDcl (p < 0.001). PLMa was higher in ADHDcl than the other groups (p < 0.001), but there were no differences in PLMa between ADHDcom and controls. ADHD children display significant alterations in their sleep patterns, and ADHDcl are more likely to have an elevated PLMa than ADHDcom. In addition, REM sleep is affected by ADHD. We postulate that ADHDcl may represent a subset of children with ADHD at high risk for hyperactivity during sleep. PMID- 12736393 TI - Defective production of IL-18 and IL-12 by cord blood mononuclear cells influences the T helper-1 interferon gamma response to group B Streptococci. AB - Human neonates are uniquely susceptible to group B streptococcal (GBS) infections. We have shown that neonatal mixed mononuclear cells have a deficiency in the production of the T helper-1 (Th-1) cytokine, interferon gamma (IFN gamma), and that incubation of neonatal neutrophils with recombinant IFN-gamma corrects these neutrophil defects. IL-12 and the more recently described IL-18 are also Th-1 type cytokines that are able to induce the production of IFN-gamma in the presence of bacteria and bacterial products. We examine the ability of GBS to induce the production of IFN-gamma, IL-18, and IL-12 by cord blood mixed mononuclear cells and compared these results with the IFN-gamma, IL-18, and IL-12 response of mixed mononuclear cells from adult blood. We demonstrate that cord blood mixed mononuclear cells produced significantly less IFN-gamma, IL-18, and IL-12 in response to GBS compared with mixed mononuclear cells from adults. Cord blood mixed mononuclear cells' production of IFN-gamma is enhanced by added recombinant IL-18 and IL-12. The maximal cord blood cell production of IFN-gamma, in response to GBS, is achieved by priming the cells with both IL-18 and IL-12. We conclude that neonatal mixed mononuclear cells exhibit deficiencies in three main Th-1 type cytokine responses, IFN-gamma, IL-12, and IL-18. This combined Th 1 type cytokine deficiency may contribute to the enhanced susceptibility of the human neonate to GBS and other microbial infections. PMID- 12736395 TI - Neonatal hyperbilirubinemia in Japanese neonates: analysis of the heme oxygenase 1 gene and fetal hemoglobin composition in cord blood. AB - Neonatal hyperbilirubinemia is frequent and severe in Japanese infants. Although the G71R mutation of the bilirubin uridine diphosphate-glucuronosyltransferase gene is associated with severe neonatal hyperbilirubinemia in this population, it accounts for only half of the neonates with severe hyperbilirubinemia. It was suggested that increased bilirubin production would also be associated with severe neonatal hyperbilirubinemia in Japanese infants. To elucidate the genetic factors causing severe hyperbilirubinemia in these patients, we studied two notable factors associated with bilirubin production: heme oxygenase-1, a key enzyme of heme metabolism, and fetal Hb composition, a factor possibly associated with heme load in neonates. We first determined the sequences of promoter and all coding regions of the heme oxygenase-1 gene in Japanese neonates who had undergone phototherapy, but found no mutation except for the polymorphic (GT)n repeats in the promoter region. These repeats modulate the transcription of the heme oxygenase-1 gene, and the longer repeat sequences are known to reduce the transcription. We detected a significant difference in the allele frequencies of each number of (GT)n repeats between Japanese and German populations. However, we could not find a relation between those polymorphisms and neonatal hyperbilirubinemia. We next analyzed the state of Hb switching of the gamma- to beta-globin chain and the phenotype of gamma-globin chain isoforms in cord blood. We found no relation between fetal Hb composition and neonatal hyperbilirubinemia. Further studies are required to elucidate genetic or environmental factors in neonatal hyperbilirubinemia in Japanese infants. PMID- 12736396 TI - A randomized controlled trial on the effect of montelukast on sputum eosinophil cationic protein in children with corticosteroid-dependent asthma. AB - Previous adult studies demonstrated the clinical efficacy of an additional treatment with leukotriene receptor antagonists on steroid-dependent asthma, but there is little knowledge about anti-inflammatory add-on effects within the lung. In this study, we hypothesized that steroid-treated children exhibit a decrease in bronchial inflammation in induced sputum under additional treatment with montelukast. Twenty-five asthmatic children aged 6 to 14 y, who had been taking inhaled corticosteroids (400-800 microg/d budesonide) regularly for at least 12 wk, were randomized to receive additional treatment with either montelukast (5 mg orally, once daily) or placebo over a 4-wk period. As primary efficacy variable, eosinophil cationic protein (ECP) in induced sputum as direct measurement of bronchial inflammation was assessed before and after treatment. To assure a baseline level of inflammation, an ECP concentration above 100 microg/L was required. Sputum eosinophil count, concentration of exhaled nitric oxide, urinary excretion of eosinophil protein X, and quality-of-life items were considered as secondary outcome variables. After treatment with montelukast, ECP in sputum was significantly reduced (montelukast: median -975 microg/L [5 to 95% confidence interval: -4295 to 583 microg/L]; placebo: 561 microg/L [-1335 to 3320 microg/L]; p < 0.01) and the quality-of-life score had significantly improved (p < 0.05) compared with placebo. Partly explained by low baseline levels, no statistically significant change in concentration of exhaled nitric oxide (p > 0.05), urinary excretion of eosinophil protein X (p > 0.05), or eosinophil count (p > 0.05) was found. In conclusion, add-on treatment with montelukast can suppress sputum ECP in children with steroid-dependent asthma, while at the same time an improvement in quality of life items occurs. PMID- 12736397 TI - Abnormal glycosylation of red cell membrane band 3 in the congenital disorder of glycosylation Ig. AB - A description is provided of the clinical presentation in an infant of the recently described congenital disorder of glycosylation type Ig, and the changes affecting glycosylation of red cell membrane band 3, the anion exchanger. It has been shown that the condition stems from a homozygous mutation within the human ortholog of yeast ALG12 gene, which encodes a dolichol-P-mannose:Man7GlcNAc2-PP dolichol alpha,1-6 mannosyltransferase of the endoplasmic reticulum. The clinical phenotype included prominent central and peripheral manifestations in the CNS. Although the infant studied had no anemia, band 3 abnormally separated into two fractions upon electrophoresis. The chemical composition of the glycans of both fractions was analyzed in detail. The fraction with low electrophoretic mobility was moderately hypoglycosylated (by 27%) and its mannose content was normal. The fraction with high electrophoretic mobility was deeply carbohydrate deficient (by 64%) and had 1 mol mannose in excess but only three residues of N acetylglucosamine. Glycophorin A was hypoglycosylated with respect to O-linked glycans. Glycosphingolipids of red cells were normal. We suggest that the incomplete biosynthesis of the N-linked glycan of band 3 was largely caused by the persistence of the 3-linked mannose residue on the 6-mannose arm of the trimannosyl moiety of the glycoprotein. It is remarkable that the changes recorded in band 3 have no clinical consequences. Band 3 alteration might serve as an additional indicator (some serum N-glycoproteins of hepatic origin are also indicative) of the congenital disorder of glycosylation type Ig. PMID- 12736398 TI - Matrix metalloproteinases in necrotising enterocolitis. AB - Elevated cytokines, especially TNF-alpha, have been implicated in the pathogenesis of necrotising enterocolitis (NEC). We have previously shown that TNF-alpha drives the production of matrix degrading enzymes, the matrix metalloproteinases (MMPs), in the gut wall. In this study we have therefore investigated the role of MMPs in the pathogenesis of NEC in neonates. Nine newborn infant nonnecrotic resected bowels with confirmed NEC were studied and 8 newborn infants with neonatal bowel obstructions were used as controls. Immunostaining was used to identify the numbers of monocytes, macrophages, neutrophils, and T cells in the tissue. We used quantitative, competitive RT-PCR to analyze the number of TNF-alpha, IFN-gamma, MMP, and TIMP mRNA transcripts and western blotting to analyze MMP and TIMP protein production. Double labeling (immunostaining and in situ hybridization) was used to identify the phenotype of MMP mRNA expressing cells. We found increased numbers of monocytes, macrophages, and neutrophils in NEC tissue compared with controls. The number of T cells was unexpectedly low in NEC as was the number of IFN-gamma transcripts in comparison with the control samples. Increased numbers of transcripts for TNF-alpha were detected in NEC tissue, as was mRNA expression and protein production for stromelysin-1 and TIMP-1 but not collagenase, gelatinases, or TIMP-2. The cellular source of stromelysin-1 in NEC was alpha-smooth muscle actin positive cells. These results suggest that stromelysin-1, which has the ability to degrade the mucosal extra-cellular matrix, may be responsible for the extensive tissue injury in infants with NEC. PMID- 12736399 TI - Prevention and control of iron deficiency anemia amongst young children. PMID- 12736400 TI - Anemia prophylaxis in adolescent school girls by weekly or daily iron-folate supplementation. AB - OBJECTIVE: To examine the benefits of anemia prophylaxis in adolescent school.girls by weekly or daily iron-folate supplementation. DESIGN: Prospective study. SETTING: Government girl schools of northeast Delhi. SUBJECTS: 2088 subjects (with hemoglobin greater than 7.9 g/dL), including 702 on daily and 695 on weekly iron-folate administration; 691 girls served as controls. RESULTS: About 85% girls were iron deficient out of which 49.3% were anemic. Weekly administration took longer time to raise hemoglobin but was effective as well as practical. Plasma ferritin estimation in girls showed rise in level in both the treated groups. CONCLUSION: Weekly administration of iron-folate was a practical and effective strategy for anemia prophylaxis in adolescent school girls. PMID- 12736403 TI - Recommendations of Polio Eradication Committee of Indian Academy of Pediatrics. PMID- 12736401 TI - Follow up of survival and quality of life in children after intensive care. AB - OBJECTIVE: To evaluate the health related quality of life of children after intensive care and to assess their long term survival. DESIGN: Prospective. SETTING: Tertiary Care Hospital Intensive Care Unit (ICU). SUBJECT AND METHODS: All patients admitted to ICU were enrolled prospectively over a period of 1 year. Children with ICU stay of less than 24 hrs, infants, readmission to ICU were however excluded. Survival was determined at the time of ICU discharge and 1 year later. Health status assessment was done with the help of the multiattribute health status classification (MAHSC), which has 6 domains; sensation, mobility, emotion, cognition, self care and pain. Assessment was done at two points of time - within 48 hours of admission to the ICU and 1 year after discharge. RESULTS: 150 children (mean age 5.68+/- 3.6 years) with a mean duration of ICU stay (5.7 +/- 5.5 days) were included in the study. The cumulative ICU mortality was 12.9%. Fifty-five (36.7% had no overall health impairment (no affected domains) preceding the present illness. There was overall health impairment (?1 affected domain) preceding the present illness in 95 of the 150 patients (63.3%). In the domain specific health status mobility was affected in 74 (49%) followed by pain 61 (41.2%), self care 56 (38.8%), sensation 29 (20%), cognition 21 (14.8%) and emotion 14(9.5%). After 1 year, overall state of health had improved or was equal to the premorbid state in 87 (75%). In domain specific health, the proportion improving or remaining unchanged varied from 75% (emotional) to 80% (cognition), 85.3% (pain) and 88.7% (mobility). The overall state of health had worsened as compared to the premorbid state in 29 (25%) majority with neurological illnesses. CONCLUSION: Quality of life in three-quarters of the patients was preserved and one year survival was favorable. Worsening was noted primarily in-patients with neurological illnesses. PMID- 12736402 TI - Thermostability of vaccines. PMID- 12736404 TI - Which patients are at risk? Evaluation of the morbility and mortality in newborn pneumothorax. AB - This study was conducted to evaluate the morbidity and mortality among the newborns hospitalized for pneumothorax. The data of 83 cases were analysed retrospectively according to gestational age, weight, underlying primary lung pathology, age of admittance, side of pneumothorax, drainage time, need for mechanical ventilation and mortality. Male: Female ratio was 1.6:1. Mean duration of admission was 63.8 hours (2 hours-20 days). 51 patients (61.4%) weighed les than 2500g and 41 patients (49.4%) were preterms. The mean weight was 2280 g (640 5170). Fifty one patients (61.4%) needed mechanical ventilation. The pnemothorax was on the right in 44 (53%), left in 21 (25.7%) and bilateral in 18 patients (21.7%). Overall 32 babies died. Among the non-survivors, 22 (68%) were preterm and there was a defined underlying lung pathology in 24 (75%). Twenty nine (90.6%) of them needed mechanical ventilation. The difference in mortality was significant in the presence of primary lung disease, low birth weight, prematurity and use of mechanical ventilation (P <005). PMID- 12736405 TI - Prevalence of antibodies to hepatitis A and hepatitis E virus in urban school children in Chennai. AB - With similar feco-oral mode of transmission of Hepatitis A and E viruses, and improving levels of personal hygiene among higher socioeconomic population, periodic surveillance on HAV/HEV exposure pattern may be of immense public health value. One such attempt was made in Tamilnadu, India by analysing the presence of antibodies to HAV and HEV in 185 healthy children of 6 months to 12 years of age. While anti HAV positivity was 96.9% by 12 years of age, anti HEV positivity fluctuated between 5.3-16.7%. The study suggests the necessity for developing a vaccine for HEV to prevent the frequent occurrence of HEV outbreaks in India, since natural HEV exposure does not bestow significant protection as observed in HAV. PMID- 12736406 TI - Prevalence of overweight and obesity in affluent adolescent girls in Chennai in 1981 and 1998. AB - To assess the prevalence of obesity and overweight in adolescent girls between 10 15 years of age, among the affluent families of Chennai--two studies are compared using body mass index (BMI) as a parameter. The first study done in the year 1981 (Group I) was compared with the second study in 1998 (Group II). Group I had 707 and group II had 610 girls. Overweight and obesity were denoted by BMI above 85th and 95th percentile respectively. Results showed a 9.6% prevalence of overweight and 6% prevalence of obesity in both studies. It was also observed that the BMI for the same age in the two study periods showed an increase from 1981 to 1998. BMI approximated the international reference values for BMI at age 13 years in the year 1998. PMID- 12736407 TI - Clinical and nutritional profile of children with celiac disease. AB - This prospective study was aimed to evaluate the clinical and nutritional profile of children diagnosed as celiac disease (CD) as per the modified ESPGAN criteria, at the time of diagnosis and after institution of gluten free diet. Out of 65 enrolled cases of CD, 7 (10.8%) children did not follow a strict dietary compliance. Only 41 children with satisfactory dietary compliance on gluten free diet (GFD) who regularly attended the follow up for at least 6 months were evaluated for their nutritional and hematological status. Results were compared with age and sex matched controls. The mean age of diagnosis was 8.7 +/- 3.3 years. Diarrhea and failure to thrive were the most common presenting symptoms. At diagnosis, the nutritional and hematological indices were significantly lower in patients than in controls. Mean duration of follow up on GFD was 22 months (range 6-48 +/- 5.6 months). On follow up, height for age Z score was significantly lower, mean BMI was significantly higher, and weight for age Z score, weight for height Z score (%), mean triceps and biceps skin fold thickness, and mid arm circumference were comparable to controls. At diagnosis, 80% cases had microcytic hypochromic anemia and 20% had dimorphic anemia. On GFD for at least a period of more than 6 months, 19% had microcytic anemia and in 81% the hematological picture was normocytic normochromic. 60% cases had thrombocytosis at diagnosis in comparison to 2.3% after treatment. Institution of GFD leads to rapid improvement in clinical picture as well as most of the nutritional and hematological parameters. PMID- 12736408 TI - Initiation of lactation and establishing relactation in outpatients. AB - The present study was conducted to evaluate whether mothers with babies less than 6 weeks of age can be helped to initiate or establish lactation in outpatient department. One thousand mothers with babies less than 6 weeks of age who completed 10 days of follow-up in outpatient have been analyzed. Mothers who had either stopped breastfeeding or were not able to initiate breastfeeding were helped with establishing lactation at outpatient clinic. Sick babies and mothers having breast problem and systemic illness were excluded. 91.6% of mothers succeeded in establishing lactation within 10 days. 83.4% mothers achieved complete lactation and 8.2% of mothers achieved partial lactation in 10 days. However, 8.4% of mothers could not be helped in this study. They had to be hospitalized because of various reasons. It is possible to help majority of mothers with lactation difficulties at the outpatient when the baby is less than 6 weeks of age. Helping mothers with proper attachment at the breast appears to be crucial for success. PMID- 12736409 TI - Pneumatosis coli, a benign form of necrotising enterocolitis. AB - Necrotising enterocolitis (NEC) is the most common acquired gastrointestinal emergency in neonates. Presence of pneumatosis intestinalis is taken as evidence of definite NEC. A distinctive but rare form of NEC called "pneumatosis coli" has been described, presenting with gross blood in stools and minimal or absent local and systemic signs. Radio-graph characteristically reveal isolated colonic pneumatosis without small bowel involvement. Pneumatosis coli has a more benign course compared with definite NEC. Total parenteral nutrition, antibiotics, an appropriate duration off feeds and close observation remain the corner stones of therapy assuring a benign course. PMID- 12736411 TI - Diamond-Blackfan anemia: report of 6 cases. AB - Diamond-Blackfan anemia is a rare congenital hypoplastic anemia. We report 6 children diagnosed as Diamond-Blackfan anemia at our clinic. All had severe pallor at presentation, with mild hepatomegaly and just palpable spleen in one child. Thumb anomaly was present in one of them. All of them had macrocytic or normocytic anemia with reticulocytopenia, and bone marrow examination revealed marked erythroid hypoplasia. All of them were treated with oral steroids with a good response. PMID- 12736410 TI - Acute renal failure due to acute tubulointerstitial nephritis. AB - Acute interstitial nephritis (AIN) should be ruled out in children with unexplained acute renal failure. We present a 4 1/2 year old girl who presented with oliguric acute renal failure preceded by a febrile illness. Renal histopathology revealed features of drug induced AIN. She recovered with dialysis, other supportive treatment and a course of steroids. PMID- 12736412 TI - Fibular hemimelia. AB - We report three cases with congenital longitudinal deficiency of fibula. Two cases were of Type IB and one was Type II. PMID- 12736413 TI - Macrodystrophia lipomatosa. PMID- 12736414 TI - Aphallia. PMID- 12736415 TI - Post injection sciatic nerve injury. PMID- 12736416 TI - Plasmodium falciparum malaria with coexisting parvovirus B19 infection. PMID- 12736417 TI - Trisomy 22 with unusual phenotype. PMID- 12736418 TI - Prevalence of night blindness amongst pregnant women of urban slum communities in Delhi: a pilot study. PMID- 12736419 TI - Indigenous visceral leishmaniasis: two cases from Kerala. PMID- 12736420 TI - In the wake of mutant virus of measles and immunization strategy. PMID- 12736421 TI - Improving oxygenation in preterm neonates with respiratory distress. PMID- 12736423 TI - Baby friendly hospitals: how friendly are they? PMID- 12736422 TI - Arthrogryposis multiplex congenita--spinal muscular atrophy association. PMID- 12736424 TI - Age-related change and allometry of skull and canine of sea otters, Enhydra lutris. AB - Skulls and canines of 460 sea otters from Lopatka Cape, Kamchatka, were examined to assess development patterns, individual variation and sexual differences. An allometric formula was applied to morphometrical data, and the relative growth of each character to total length of skull was analyzed. In both sexes, most morphometrical characters ceased growth at about 2 years of age. Canine root length increased rapidly during the first year of life, while crown length decreased due to remarkable wear. There was large individual variation in the feeding and breathing/sniffing apparatus, while there was little variation in braincase size. There were sexual differences in most characteristics, although males and females showed similar growth patterns. The coronoid process of the mandible showed positive allometry in both sexes, and we attributed this finding to feeding habits. The fact that only male mastoids showed positive allometry may be due to the need for male otters to maintain a passing territory. PMID- 12736425 TI - Migration of erythroblastic islands toward the sinusoid as erythroid maturation proceeds in rat bone marrow. AB - It has been hitherto considered that mature erythroblasts migrate toward the sinusoid along the cytoplasmic processes of macrophages of erythroblastic islands in bone marrow. Our previous report, however, has demonstrated the morphological features of a mature erythroblastic island passing through the sinusoidal endothelium. In this study, the possibility of migration of erythroblastic islands toward the sinusoid was examined in rat bone marrow by light microscopical histoplanimetry. As a result, the more mature erythroblasts were not regularly arranged in the peripheral direction of the erythroblastic islands. Immature erythroblasts were populated more in the erythroblastic islands away from the sinusoid than in those islands neighboring the sinusoid, whereas mature erythroblasts were more in erythroblastic islands neighboring the sinusoid. These findings suggest that the formation of erythroblastic islands occurs in a region away from the sinusoid, and that erythroblastic islands migrate towards the sinusoids as erythroid maturation proceeds. PMID- 12736426 TI - Epidemiological aspects of the first outbreak of Baylisascaris procyonis larva migrans in rabbits in Japan. AB - Larva migrans caused by the common raccoon ascarid, Baylisascaris procyonis, is a zoonotic disease of critical importance in North America. Recently we encountered the first proven outbreak of this disease in Japan in domestic rabbits (Oryctolagus cuniculus) in a small wildlife park. In this park, raccoons (Procyon lotor) had been kept for 9 years, and one raccoon was donated to the park by a pet owner 8 weeks prior to the occurrence of an outbreak in rabbits. Of 12 total raccoons, three raccoons including the donated one shed B. procyonis eggs in the feces, and two of these positive raccoons were kept in metal mesh cages on wooden pedestals, 2 m distant from the rabbit enclosure. Circumstantial evidence indicates that the donated raccoon is the likely source of this outbreak. Treatment of the raccoons with an ascaricide and decontamination by extensive flaming of the cages and the contaminated dirt floor of the park achieved a transient disappearance of ascarids from all 12 enclosed raccoons. Three months after the control measures began, recurrent ascarid infection was detected in three young raccoons of less than 1.5 years of age. The potential risk of serious zoonosis by B. procyonis as well as the difficulty in a clearance of contaminated areas should be considered by pet owners and public health workers in Japan. PMID- 12736427 TI - Application of timed artificial insemination protocols to grazing Japanese black cattle with long open period. AB - To investigate the viability of timed artificial insemination (TAI) protocols in grazing Japanese Black cattle with long open period, ovarian status and progesterone and estradiol-17 beta profiles of the animals during the protocol were monitored. In 1998, prosta-glandin F(2a) (PG) was administered to 36 animals seven days after GnRH injection. Three out of the 36 animals were inseminated after detection of estrus and did not receive further treatment. The second GnRH was injected to the remaining 33 animals 48 hr after the PG injection and TAI was performed 24 hr later. In 1999, PG was injected to 25 animals six days after GnRH, the second GnRH was injected to 22 animals 48 hr after PG, and TAI was performed 16 hr later (The other three animals were inseminated before the time of TAI). The percentage of the animals with at least one functional corpus luteum and one follicle equal to or greater than 10 mm in diameter at PG injection was similar between the groups in 1998 and 1999. Likewise, the hormonal profiles were similar between the two groups. Pregnancy rates (PR) after the TAI protocols and natural mating in 1998 and 1999 were 75.0% and 88.0%, respectively. These figures were comparable to the PR obtained by conventional estrus synchronization protocols using PG (in 1995; 69.4%) or CIDR (in 1996; 59.1%). In conclusion, the TAI protocol can be applicable into grazing Japanese Black cattle with long open period. PMID- 12736428 TI - The plasma superoxide scavenging activity in canine cancer and hepatic disease. AB - To clarify the relationship between plasma antioxidant activity and diseases in dogs, plasma samples were collected from 6 healthy dogs and 16 diseased dogs (6 dogs with cancer, 5 dogs with hepatic disease, and 5 dogs with inflammation ), and measured superoxide anion scavenging activities. Antioxidant activities of canine plasma were evaluated by measuring their superoxide anion (O(2)(-.)) scavenging activities with electron spin response spectroscopy combined with spin trapping reagent, 5,5-dimethyl-1-pyrroline-N-oxide (DMPO). Total O(2)(-.) scavenging activities in the presence of plasma of diseased dogs tended to be higher than those in healthy controls, especially significant higher activities in the presence of canine plasma of hepatic disease and inflammation were observed. In diseased dogs, KCN-insensitive activities, suggesting the activity of manganese-containing superoxide dismutase (Mn-SOD), were significantly higher than those in healthy controls. Therefore, it seems that there is a possibility of utilizing of plasma O(2)(-.) scavenging activity as one of clinical indicators for oxidative-related diseases such as cancer, hepatic disease and inflammation in dogs. PMID- 12736429 TI - The relationship between cellular radiosensitivity and radiation-induced DNA damage measured by the comet assay. AB - The relationship between deoxyribonucleic acid (DNA) damage and the cell death induced by gamma-irradiation was examined in three kinds of cells, Chinese hamster ovary fibroblast CHO-K1, human melanoma HMV-II and mouse leukemia L5178Y. Cell survival was determined by a clonogenic assay. The induction and rejoining of DNA strand breaks induced by radiation were measured by the alkaline and neutral comet assays. L5178Y cells were the most radiosensitive, while CHO-K1 cells and HMV-II cells were radioresistant. There was an inverse relationship between the survival fraction at 2 Gy (SF2) and the yield of initial DNA strand breaks per unit dose under the alkaline condition for the comet assay, and also a relationship between SF2 and the residual DNA strand breaks (for 4 hr after irradiation) under the neutral condition for the comet assay, the latter being generally considered to be relative to cellular radiosensitivity. In the present analysis, it was considered that the alkaline condition for the comet assay was optimal for evaluating the initial DNA strand breaks, while the neutral condition was optimal for evaluating the residual DNA strand breaks. Since the comet assay is simpler and more rapid than other methods for detecting radiation-induced DNA damage, this assay appears to be a useful predictive assay for evaluating cellular clonogenic radiosensitivity of tumor cells. PMID- 12736430 TI - Immune responses of immunocompetent and immunocompromised mice experimentally infected with Bartonella henselae. AB - The aim of this study is to understand host immune responses in immunocompetent and immunocompromised mice against Bartonella henselae infection. BALB/c and nude (BALB/c nu/nu) mice were inoculated intraperitoneally with 10(8) colony forming units of B. henselae (Houston-1 strain). Blood, brain, liver, spleen, kidney and bone marrow samples were collected 0, 3, 7, 14, 21 and 28 days after infection and submitted to bacteriological, serological and genetical examinations. B. henselae was isolated only from the liver 3 days after infection. DNA of the inoculums was detected by polymerase chain reaction from blood, liver, and spleen samples collected from BALB/c and blood from nude mice 3 and 7 days after infection. No bacterial DNA was detected from both BALB/c and nude mice thereafter during 4 weeks observation periods. These results indicate that the T cell may not participate in the effective elimination of the organisms from mice. In addition, western blot analysis revealed that the antigens of 27.3- and 31.5 kDa reacted with IgM antibodies from the blood of BALB/c and nude mice after 3 days of infection, suggesting that these antigens were recognized by thymus independent mechanism. Furthermore the antigens were detected from the culture supernatants of B. henselae, indicating that these antigens were secreted from the organisms. PMID- 12736431 TI - Functional changes of the LH-immunoreactive cells in adenohypophysis of the common carp (Cyprinus carpio) from rivers contaminated with estrogenic chemicals. AB - The adverse effect of estrogenic chemicals on luteinizing hormone-immunoreactive (LH-ir) cells in the adenohypophysis of common carp (Cyprinus carpio) was examined using immunocytochemical and morphometric methods. Adult male fish were collected from two contaminated sites (Ishizu and Wada Rivers) and from a control pond at Agriculture, Food and Environmental Sciences, Research Center of Osaka Prefecture. The concentration of nonylphenol, bisphenol A and 17beta-estradiol in the Ishizu River was 3-4 times higher than that in the Wada River. The proportion and size of LH-ir cells were evaluated using the point-counting method by optical microscopy. In control carp, the proportion of LH-ir cells in the breeding season was significantly lower than in the pre- and post-breeding seasons. The same tendency was also found in Ishizu and Wada River carp, but without statistical significance. The proportion of LH-ir cells in Ishizu River carp was significantly lower than that of the control and the Wada River in all seasons. The LH-ir cells in control carp increased in size in the breeding season. LH-ir cells in Ishizu River carp were significantly (p<0.05) smaller than those in control fish, but not different from Wada River carp. A disturbance in the secretory function of LH-ir cells was found in carp from the Ishizu River; granulation and vacuolation were not in synchronization with those of control and Wada River fish. Our data suggest that the estrogenic chemicals in the Ishizu River interfere with functions of LH-ir cells directly or through the testis. PMID- 12736432 TI - Phenotype and function of murine discrete Peyer's patch macrophage derived - dendritic cells. AB - The phenotype and function of peritoneal cavity macrophage-derived dendritic cells (PEC-DC) was previously reported. In this study we have gone further in using our established culture system to generated discrete Peyer's patch dendritic cells (DPP-DC) from murine discrete Peyer's patch macrophages (DPP-Mo), following stimulation with granulocyte macrophage colony stimulating factor (GM CSF) plus interleukin 4 (IL-4) for 7 days. DPP-Mo from murine small intestines were obtained by mechanical disruption of discrete Peyer's patches (DPP), followed by metrizamide density gradient centrifugation to remove Peyer's patch resident DC and debri, after which an overnight adherent step in tissue culture medium was carried out for macrophage enrichment. Characterization of the generated DPP-DC was carried out using well-established criteria of morphology, expression of membrane antigens and capacity for antigen presentation. Dendritic cells expressed DEC-205, F4/80 and CD34 at high levels, but exhibited very low CD11c levels. They were shown to present soluble protein antigen to CD3(+) spleen T cells. A comparison of the surface antigen expression in the progenitor DPP-Mo population and the generated DPP-DC showed a significant decrease in MHC class II levels and a marked down regulation of the co-stimulatory molecule CD86 (B7-2). High expression of the haemopoietic progenitor marker CD34 indicates that the generated DC, possess a haemopoietic rather than myeloid origin. Taken together, these results may provide a better understanding of the complex network regulating mucosal immune responses. PMID- 12736433 TI - Identification and characterization of cytopathogenic Mycoplasma hyorhinis from swine farms with a history of abortions. AB - A virus-like cytopathic agent isolated from swine farms with a history of recurrent abortion episodes was investigated. We employed a differential display reverse transcription-polymerase chain reaction (ddRT-PCR) to obtain genetic information of the cytopathic agent. Partial nucleotide sequence (527 bp) obtained from differentially displayed PCR fragments showed 88.7% similarity with the 23S rRNA gene of Mycoplasma hyopneumoniae. Unexpectedly, the 5' portion (1 333 bp) of the sequence shared 96.1% similarity with 5' untranslated region (UTR) of human prostate tumor inducing gene 1 (PTI-1). Cytopathic effects and extranuclear DNA fluorescence were no longer observed when BM-cyclin was added in the culture medium, suggesting that BM-cyclin sensitive mycoplasma-like organisms caused the cell death. Further evidence supporting the cytopathic agent as a mycoplasma-like organism was obtained by the capability of (3)H-thymidine and (3)H-uridine incorporation, a single peak in buoyant density gradient profile (1.20-1.24 g/ml), and ultrastructural morphology. Unlike M. hyopneumoniae, the organism was not propagated in Friis medium. Nucleotide sequence of 16S rRNA obtained from the cytopathic agent showed 0.8-1.0% divergences with other M. hyorhinis strains, suggesting that the newly isolated cytopathogenic swine mycoplasma was a variant form of M. hyorhinis. Striking homology between a portion of the 23S rRNA gene of M. hyorhinis and 5' UTR of human PTI-1 implicated that M. hyorhinis might potentially be related to the evolution of human PTI-1. PMID- 12736434 TI - Interleukin-6 protects skin lesion caused by 7,12-dimethylbenz[a]anthracene. AB - Interleukin-6 (IL-6) regulates essential physiological functions such as acute phase reaction, immune response and hematopoiesis. We here studied possible protective effects of IL-6 on skin damage caused by 7,12 dimethylbenz[a]anthracene (DMBA) by using IL-6 null mice. The mice were topically applied with a single dose of DMBA (500 microg /mouse) on the dorsal skin. Osmotic pumps filled with recombinant human IL-6 (rhIL-6) were implanted subcutaneously on the ventral side of the mice. Control mice received PBS instead of rhIL-6 by the pump. Severe skin damage was observed in IL-6-null mice, whereas only epidermal hyperplasia was observed in the wild-type mice. Recombinant hIL-6 treatment to DMBA-treated IL-6-null mice suppressed the occurrence of the skin damage, indicating. PMID- 12736435 TI - Delayed-type hypersensitivity in sheep induced by synthetic peptides of bovine leukemia virus encapsulated in mannan-coated liposome. AB - Four sheep were immunized with synthesized peptides, derived from bovine leukemia virus (BLV) envelope glycoprotein, encapsulated in mannan coated liposomes as adjuvant. On the seventh day after the immunization, the sheep were intradermally challenged with BLV antigen, or synthesized peptides. The areas challenged with antigen were increased skin thickness and biopsied sequentially for immunohistological examinations. Strong delayed-type hypersensitivity was induced in sheep immunized and challenged with peptides encapsulated in mannan-coated liposomes. The major phenotype of the infiltrating lymphocytes was CD5(+). The ratio of CD4(+) to CD8(+) cells was about 1:1. PMID- 12736436 TI - cDNA cloning of a Rab5 homologue in Babesia gibsoni. AB - For studying protein trafficking in Babesia-infected erythrocyte, we describe the cloning of a Rab5, one of molecular marker for vesicle trafficking in eukaryotic cells, gene homologue in Babesia gibsoni (BgRab5). The full-length cDNA of BgRab5 is 1,020 bp long with an open reading frame encoding a protein of 220 amino acids. The deduced amino acid sequence of BgRab5 contained the highly conserved GTP-binding consensus sequence and shares about 40% homology with that of Rab5 from Plasmodium falciparum, Toxoplasma gondii, Dog, Lotus japonicusor, Oryza sativa. Northern blot analysis showed that the BgRab5 probe hybridized with a 1kb band in total RNA from parasitized erythrocytes, that was consistent with the size of the BgRab5 full-length cDNA. PMID- 12736437 TI - Steroid injection therapy in a feline solitary bone cyst. AB - A nine months old Japanese domestic cat with a solitary bone cyst, which is a benign tumor-like lesion and is particularly uncommon in feline practice was referred. Radiographic examination revealed an expansive radiolucency in the distal metaphysis of the right ulna and pathologic fracture. The histological examination demonstrated immature osteogenesis consisting of osteoblasts surrounded by connective tissue. We applied drainage and instillation of steroid solution into the cystic cavity. Clinical signs, including lameness and pain, disappeared three weeks after the therapy started. Fourteen months after the therapy, the cystic lesion of bone was remodeling successfully without re developing. We conclude that our procedure was useful treatment in this case. PMID- 12736438 TI - Equine pyoderma associated with malnutrition and unhygienic conditions due to neglect in a herd. AB - Twelve horses kept at a riding club suffered from pyoderma. All the horses displayed crusting, scaling and alopecia. The lesions were distributed in the chest, back, rump and limbs. Some of the horse patients also showed epilation with an attached crust similar to a 'paintbrush lesion' of dermatophilosis, but normal skin flora or opportunistic pathogenic bacteria were only isolated from the lesions. Some patients clearly showed weight loss, anemia and low levels of serum protein and cholesterol. General condition and skin lesions of the patients were improved gradually with improvement of feed and environment after being moved to new stables. Malnutrition under conditions of poor hygiene and poor management due to neglect might be associated with these equine cases of pyoderma in the herd. PMID- 12736439 TI - Ebstein's anomaly in a beagle dog. AB - An intact male beagle dog aged 1 year was referred because of shortness of breath, exercise intolerance and cardiac murmur. Based on the results from electrocardiography, thoracic radiography and echocardiography, the dog was diagnosed as Ebstein's anomaly. Although the orally administered digoxin, vasodilators and diuretics partially improved congestive signs, the dog became to be refractory and died 20 months after the diagnosis. Necropsy confirmed malformation and apical displacement of the basal attachment of tricuspid valve leaflets. PMID- 12736440 TI - Phagocytosis of bovine blood and milk polymorphonuclear leukocytes after ozone gas administration in vitro. AB - To determine the effects of ozone on the phagocytosis of bovine polymorphonuclear leukocytes (PMNs), ozone gas was administered in vitro on the blood and milk of healthy lactating cows, cows with acute mastitis, and cows with milk fever. In the blood of healthy dairy cattle, although there was no significant effect of ozone gas on the viability of the leukocytes, phagocytosis of PMNs significantly decreased. In contrast, ozone gas administration in vitro significantly increased phagocytosis of PMNs from the blood of cows with acute mastitis and milk fever, and from mastitic milk. These findings showed that ozone administration in vitro has positive and negative effects on bovine PMN phagocytosis, depending on the health status of the animal. PMID- 12736441 TI - Postnatal development of the mouse volatile papilla taste bud cells. AB - In the present study, we examined specific markers for taste bud cells in the mouse and the postnatal development of volatile papilla taste bud cells in ddY mice. We examined the immunoreactivity of 4 types of carbonic anhydrase isoenzymes, CA I, CA II, CA III and CA VI, as specific markers for taste bud cells, and K8.13 cytokeratin antibody as a specific marker for the lingual epithelial cells. Of the carbonic anhydrase isoenzymes, only CA III immunoreactivity was clearly detected in the spindle shaped gustatory cells. CA VI immunoreactivity was detectable in suspentacular cells. CA I and CA II antibodies did not recognize any taste bud cell specifically. K8.13 cytokeratin immunoreactivity was detected in the lingual epithelial cells, but not in taste bud cells. At 7 days after birth, the suckling phase, very small taste buds developed from the anaplastic gustatory cells. At 14 days after birth, the taste buds showed larger size than those at 7 days after birth. At 21 days birth, after the weaning phase, taste bud structure approximated the mature structure. These results demonstrate the specificity of anti-CA III and anti-CA VI for gustatory cells and suspentacular cells, respectively. These markers should be useful for an analysis of taste bud development in mice. PMID- 12736442 TI - Evaluation of feline serum amyloid A (SAA) as an inflammatory marker. AB - The concentration of feline serum amyloid A (fSAA) was determined by a direct enzyme-linked immunosorbent assay (ELISA) by using fSAA specific monoclonal antibodies, to evaluate the fSAA as an inflammatory marker in cats. The mean concentration +/- standard deviation of fSAA was found to be 0.60 +/- 1.06 microg/m l and 33.65 +/- 67.59 microg/ml in serum samples from normal cats (n=45) and cats (n=312) with various diseases and disorders, respectively. A significant difference (p<0.001) was found between the two groups. It was also found that the concentration of fSAA begins to increase rapidly at approximately 3-6 hr after spay, and increases up to significantly high levels in some disorders, like injury, renal failure, infectious diseases, etc. PMID- 12736444 TI - Dammarane triterpenes of Commiphora confusa resin. AB - Fractionation of a steam distilled residue of Commiphora confusa resin has yielded four novel dammarane triterpenes characterised as (20S)-3beta-acetoxy 12beta,16beta-trihydroxydammar-24-ene, (20S)-12beta,16beta-trihydroxydammar-24 ene-3beta-O-beta-glucopyranoside, (20S)-3beta-acetoxy-12beta,16beta,25 tetrahydroxydammar-23-ene, and (20S)-3beta,12beta,16beta,25-pentahydroxydammar-23 ene. The known compounds beta-amyrin, 3beta-amyrinacetate, 2 methoxyfuranodienone, 2-acetoxyfuranodienone, (20R)-3beta-acetoxy-16beta dihydroxydammar-24-ene, (20R)-3beta,16beta-trihydroxydammar-24-ene, 3beta-acetoxy 16beta-hydroxydammar-24-ene, 3beta-hydroxydammar-24-ene, 3beta-acetoxydammar-24 ene, and beta-sistosterol were also isolated from the same extract. The structures of the compounds were determined using spectroscopic, physical, and chemical methods. PMID- 12736443 TI - Further bisabolenes and dammarane triterpenes of Commiphora kua resin. AB - From the resins of Commiphora kua a novel bisabolene; 6-hydroxy-2-methyl-5-(5' hydroxy-1'(R),5'-dimethylhex-3'-enyl)-phenol together with two new dammarane triterpenes, 3beta,16beta,20(S),25-tetrahydroxydammar-23-ene and 3beta-acetoxy 16beta,20(S),25-trihydroxydammar-23-ene, have been isolated. In addition, being reported are known compounds identified as 2-methyl-5-(4'(S)-hydroxy-1'(R),5' dimethylhex-5'-enyl)-phenol, 2-acetoxyfuranodienone, 2-methoxyfuranodienone, 3beta,16beta,20(R)-trihydroxydammar-24-ene and its acetate derivative, 3beta acetoxy-16beta,20(R)-dihydroxydammar-24-ene, and beta-amyrin and its acetate derivative. 2-Methyl-5-(4'(S)-hydroxy-1'(R),5'-dimethylhex-5'-enyl)-phenol displayed fungicidal activity against Cladosporium cucumernum on TLC assay. PMID- 12736445 TI - Effect of crystallization behavior of polyethylene glycol 6000 on the properties of granules prepared by fluidized hot-melt granulation (FHMG). AB - The objective of this study was to investigate the effect of the crystallization behavior of Macrogol 6000 (polyethylene glycol 6000; PEG 6000), used as a binder, during the solidification process on the properties of mononucleic granules prepared by the fluidized hot-melt granulation (FHMG) technique. Crystallization of PEG 6000 from molten liquid was investigated using differential scanning calorimetry (DSC) and hot stage microscopy. The results obtained from the measurement of isothermal crystallization demonstrated that crystallization of PEG 6000 was either slow or rapid. Analysis based on solid-state decomposition showed that slow crystallization was due to the two-dimensional growth of nuclei mechanism, while rapid crystallization was due to the three-dimensional growth of nuclei mechanism. Observation of the crystallization of PEG 6000 by hot stage microscopy supported the existence of two different crystallization mechanisms. Granules containing PEG 6000 that underwent rapid crystallization during FHMG showed a significantly higher fraction powder under 150 microm in diameter. This was caused by the loss of powder particles from the surface of mononucleic granules during the solidification process, because many cracks were observed after crystallization of PEG 6000 with a short isothermal crystallization time (ICT) due to the reduced of sticking of particles. The results of this study suggested that the crystallization behavior of the binder during the solidification process of FHMG can influence the properties of the resultant granules, such as particle size distribution, content uniformity or taste masking. It was also indicated that measuring the ICT using DSC was a useful method to classify PEG 6000. PMID- 12736446 TI - Synthesis, characterization, and antibacterial activities of some rare Earth metal complexes of pipemidic acid. AB - Eight new solid complexes of pipemidic acid (PPA) with trichlorizated rare earth metals LaCl(3), CeCl(3), PrCl(3), NdCl(3), SmCl(3), TbCl(3), DyCl(3), and YCl(3) have been synthesized. The complexes were characterized by elemental analyses, IR, NMR, and molar conductance measurements. The general formulas of the complexes are [M(PPA)(4)]Cl(3) (M=Ce(III), Pr(III), Nd(III), Sm(III), Tb(III), Dy(III), Y(III)), and [La(PPA)(4)Cl]Cl(2). At the same time, the antibacterial activities of PPA and four of its complexes were tested. The results show that PPA and its complexes all have inhibitory action against bacteria of Escherichia coli, Bacillus subtilis, Streptococcus pneumoniae, and Pseudomonas aeruginosa but not Staphylococcus aureus. We compared their antibacterial activities and found that the antibacterial activity of [La(PPA)(4)Cl]Cl(2) against S. pneumoniae is much stronger than that of PPA and the other complexes. PMID- 12736448 TI - Dioxopyrrolines. LXII. Diels-Alder reaction of 1-Aryl-4- and 5-methoxycarbonyl-1H pyrrole-2,3-diones with various 1,3-dienes. AB - Two new dioxopyrrolines (1-aryl-4-methoxycarbonyl-1H-pyrrole-2,3-dione 6 and the 5-methoxycarbonyl isomer 8) behaved as good dienophiles to some kind of 1,3 dienes examined. In most cases, the products were explained by the reaction where the largest lobe of HOMO of dienes reacted to the larger LUMO of dienophiles in an expected cis-endo manner. However, in the reactions of 8 with alkylbutadienes, piperylene and isoprene, abnormality in the reaction was observed, which was well explained by taking account of steric factors. PMID- 12736447 TI - Sinodielides E-H, four new guaianolides, from the root of Sinodielsia yunnanensis. AB - Four new guaianolides, sinodielides E-H (1-4), were isolated from the root of Sinodielsia yunnanensis WOLFF. Their structures were established by spectral evidence. PMID- 12736449 TI - Biotransformation of pinoresinol diglucoside to mammalian lignans by human intestinal microflora, and isolation of Enterococcus faecalis strain PDG-1 responsible for the transformation of (+)-pinoresinol to (+)-lariciresinol. AB - By anaerobic incubation of pinoresinol diglucoside (1) from the bark of Eucommia ulmoides with a fecal suspension of humans, eleven metabolites were formed, and their structures were identified as (+)-pinoresinol (2), (+)-lariciresinol (3), 3'-demethyl-(+)-lariciresinol (4), (-)-secoisolariciresinol (5), (-)-3-(3", 4" dihydroxybenzyl)-2-(4'-hydroxy-3'-methoxybenzyl)butane-1, 4-diol (6), 2-(3', 4' dihydroxybenzyl)-3-(3", 4"-dihydroxybenzyl)butane-1, 4-diol (7), 3-(3" hydroxybenzyl)-2-(4'-hydroxy-3'-methoxybenzyl)butane-1, 4-diol (8), 2-(3', 4' dihydroxybenzyl)-3-(3"-hydroxybenzyl)butane-1, 4-diol (9), (-)-enterodiol (10), ( )-(2R, 3R)-3-(3", 4"-dihydroxybenzyl)-2-(4'-hydroxy-3' methoxybenzyl)butyrolactone (11), (-)-(2R, 3R)-2-(3', 4'-dihydroxybenzyl)-3-(3", 4"-dihydroxybenzyl)butyrolactone (12), (-)-(2R, 3R)-3-(3"-hydroxybenzyl)-2-(4' hydroxy-3'-methoxybenzyl)butyrolactone (13), 2-(3', 4'-dihydroxybenzyl)-3-(3" hydroxybenzyl)butyrolactone (14), 2-(3'-hydroxybenzyl)-3-(3", 4" dihydroxybenzyl)butyrolactone (15) and (-)-(2R, 3R)-enterolactone (16) by various spectroscopic means, including two dimensional (2D)-NMR, mass spectrometry and circular dichroism. A possible metabolic pathway was proposed on the basis of their structures and time course experiments monitored by thin-layer chromatography. Furthermore, a bacterial strain responsible for the transformation of (+)-pinoresinol to (+)-lariciresinol was isolated from a human fecal suspension and identified as Enterococcus faecalis strain PDG-1. PMID- 12736450 TI - Comparative molecular field analysis study of stilbene derivatives active against A549 lung carcinoma. AB - A series of 43 stilbene derivatives that showed cytotoxicity against human lung carcinoma (A549) was analyzed using comparative molecular field analysis (CoMFA) for defining the hypothetic pharmacophore model. The polyoxylated stilbenes were found to be active inhibitors of tubulin polymerization. Several cis-stilbenes are structurally similar to combretastatins. However, the trans-stilbenes are assumed to be close to resveratrol found in grapes and have been reported to be potential cancer chemopreventive agents by modulating the initiation, promotion, and progression of the carcinogenic process. With several synthesized compounds that were evaluated for antitumor cytotoxicity against human lung tumor cells (A549), the stilbene derivatives were subjected to CoMFA. To perform systematic molecular modeling of these compounds, a conformational search was carried out based on the precise dihedral angle analysis of the lead compound (1p). The X-ray crystallographic structure of combretastatin A-1 was also used for defining the active conformers of the compounds. After determining the energy-minimized conformers of the lead compound (1p), CoMFA was performed using five different alignments. The three dimensional (3D)-quantitative structure-activity relationship study resulted in reasonable cross-validated, conventional r(2) values equal to 0.640 and 0.958, respectively. PMID- 12736451 TI - Synthesis, conformational analysis and free radical scavenging activity of some new spiropyranoquinolinones. AB - A series of novel spiroadamantyl- and spirocyclical substituted pyranoquinolin-2 ones were synthesized and the conformation of the pyran ring was investigated. The free radical scavenging activity of the synthesized compounds was determined by their interaction with the stable free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH). All compounds tested scavenged the DPPH radical and among them derivatives possessing extended conjugation showed the highest activity. PMID- 12736452 TI - Minor withanolides from Physalis philadelphica: structures, quinone reductase induction activities, and liquid chromatography (LC)-MS-MS investigation as artifacts. AB - As a result of a bioactivity-guided search for novel, plant-derived cancer chemopreventive agents, ixocarpalactone A (5) was isolated previously as a potent quinone reductase inducer from the leaves and stems of Physalis philadelphica. In the present study, this promising lead compound was reisolated in gram quantities for in vivo biological testing. During the course of this work, four additional minor new withanolides were also obtained and characterized, namely, 2,3-dihydro 3beta-methoxyixocarpalactone A (1), 2,3-dihydro-3beta-methoxyixocarpalactone B (2), 2,3-dihydroixocarpalactone B (3), and 4beta,7beta,20R-trihydroxy-1-oxowitha 2,5-dien-22,26-olide (4). However, compounds 1 and 2 were determined using liquid chromatography (LC)-MS-MS to be artifacts generated during the extraction and isolation procedure. Ixocarpalactone A was detected in the fresh fruits (tomatillos) of P. philadelphica by LC-MS-MS analysis at a concentration of 143+/ 4.53 ppb. PMID- 12736453 TI - Diastereoselective solid-phase radical addition to oxime ether anchored to polymer support. AB - Stereocontrol in radical reactions of oxime ether anchored to polymer support was studied. Highly diastereoselective solid-phase radical reaction was achieved by using triethylborane and diethylzinc as a radical initiator at low reaction temperature, providing a novel method for the synthesis of the alpha-amino acid derivatives with excellent diastereoselectivities. PMID- 12736454 TI - Five new sesquiterpenoids and a new diterpenoid from Erigeron annuus (L.) PERS., Erigeron philadelphicus L. and Erigeron sumatrensis RETZ. AB - The aerial parts of Erigeron annuus (L.) PERS., E. philadelphicus L. and E. sumatrensis RETZ. (Compositae) have been investigated chemically. A new sesquiterpenoid, 6beta,14-epoxyeudesm-4(15)-en-1beta-ol (1), and a new diterpenoid, philadelphinone (6), have been isolated from E. philadelphicus. Four new sesquiterpenoids, (7R*)-opposit-4(15)-ene-1beta,7-diol (2), 11-methoxyopposit 4(15)-en-1beta-ol (3), 15-methoxyisodauc-3-ene-1beta,5alpha-diol (4) and 10alpha hydroxycadin-4-en-15-al (5), have been isolated from E. annuus. Compounds 2 and 4 were also isolated from E. sumatrensis. The structures of the new compounds were elucidated on the basis of their spectral data. PMID- 12736455 TI - Analysis of the peptides (Prp106-126, MSI-78A, and Oxaldie 1) with the same biological activity by discrete Fourier transform: toward a selection rule in ligand-receptor interaction. AB - Here, we first report a novel method in which the "desired cross-spectrum" of the peptides Prp106-126, MSI-78A, and oxaldie 1 with the same biological activities is obtained by the multiplication of two cross-spectra derived from the RNA sequence and from the cognate amino acid sequence by discrete Fourier transform (DFT), respectively. Based on a well-known method reported previously, we investigated the cross-spectrum by the multiplication of two of three desired cross-spectra. As a result, we found that one prominent peak occurring in the three cross-spectra showed the same frequency when a binary scale was used as a parameter of nucleotide or amino acid in the analysis. Moreover, we examined the relationship between a binary scale and other physicochemical ones. Almost the same results could be reproduced when the absolute electronegativity scale (or the absolute hardness one) was used, but not in the case of the hydrophobic or electron-ion interacting potential scale reported previously. This indicates that either the absolute electronegativity scale (or the absolute hardness one) or a binary scale, or both is very useful in extracting the information desired for various proteins by the present method from the amino acid and the RNA sequence. PMID- 12736456 TI - Four new 2-(2-phenylethyl)chromone derivatives from withered wood of Aquilaria sinensis. AB - Four new chromone derivatives, 5-hydroxy-6-methoxy-2-(2-phenylethyl)chromone (1), 6-hydroxy-2-(2-hydroxy-2-phenylethyl)chromone (2), 8-chloro-2-(2-phenylethyl) 5,6,7-trihydroxy-5,6,7,8-tetrahydrochromone (3), 6,7-dihydroxy-2-(2-phenylethyl) 5,6,7,8-tetrahydrochromone (4) were isolated from the MeOH extract of withered wood of Aquilaria sinensis, together with seven known constituents of agarwood. PMID- 12736457 TI - Synthesis and biological activity of novel 5-(omega-aryloxyalkyl)oxazole derivatives as brain-derived neurotrophic factor inducers. AB - A novel series of 5-(omega-aryloxyalkyl)oxazole derivatives was prepared and their effects on brain-derived neurotrophic factor (BDNF) production were evaluated in human neuroblastoma (SK-N-SH) cells. Syntheses were performed by construction of an oxazole ring as a key reaction. Most of the 5-(omega aryloxyalkyl)oxazole derivatives markedly increased BDNF production in SK-N-SH cells. 4-(4-Chlorophenyl)-2-(2-methyl-1H-imidazol-1-yl)-5-[3-(2 methoxyphenoxy)propyl]-1, 3-oxazole, one of the most promising compounds, showed potent activity (EC(50)=7.9 microM) and the improvement of the motor nerve conduction velocity and the tail-flick response accompanied by a recovery of the brain-derived neurotrophic factor level in the sciatic nerve of streptozotocin (STZ)-diabetic rats. PMID- 12736458 TI - New triterpenoid saponins from Stelmatocrypton khasianum. AB - Four new triterpenoid saponins, designated as stelmatotriterpenosides E-H (1-4), together with three known compounds, asterbatanoside B (5), 2alpha,3beta,19alpha,23-tetrahydroxy-olean-12-en-28-oic acid-3-O-beta-D glucopyranosyl-28-O-beta-D-glucopyranosyl ester (6) and 2alpha,3beta,19alpha,23 tetrahydroxy-urs-12-en-28-oic acid-3-O-beta-D-glucopyranosyl-28-O-beta-D glucopyranosyl ester (7), were isolated from the stems of Stelmatocrypton khasianum. On the basis of chemical and spectral evidence, the structures of 1-4 were established as 2alpha,3beta,23-trihydroxy-olean-12-en-28-oic acid-3-O-beta-D glucopyranosyl-28-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl ester (1), 2alpha,3beta,23-trihydroxy-urs-12-en-28-oic acid-3-O-beta-D-glucopyranosyl 28-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl ester (2), 2alpha,3beta,19alpha-trihydroxy-urs-12-en-28-oic acid-3-O-beta-D-glucopyranosyl 28-O-beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosyl ester (3), and 2beta,3beta,19alpha-trihydroxy-urs-12-en-24,28-dioic acid-24-O-beta-D glucopyranosyl-28-O-beta-D-glucopyranosyl diester (4). PMID- 12736459 TI - Fluorescence studies on the interactions of barbaloin with bovine serum albumin. AB - The fluorescence quenching reactions of barbaloin with bovine serum albumin (BSA) in pH 7.20 Tris-HCl buffer solution were studied. The quenching mechanism of BSA by barbaloin was interpreted using the Stern-Volmer (S-V) mechanism. The binding constant K values were 2.78 x 10(5) (293 K), 1.87 x 10(5) (310 K), 1.25 x 10(5) (318 K), and the number of binding sites (n) were 1.18, 1.14, and 1.09, respectively. In addition, the thermodynamic functions enthalpy (deltaH degrees ) and entropy (deltaS degrees ) for the reaction were also calculated according to Vant's Hoff equation were -23.7 kJ/mol and 23.6 J/mol, respectively. Plausible explanations of the quenching mechanism are discussed on the basis of a hydrophobic interaction between barbaloin and BSA. PMID- 12736460 TI - Iridoid esters from Patrinia saniculaefolia. AB - Two new iridoids, named patridoid I (1) and patridoid II (2), were isolated from the whole plant of Patrinia saniculaefolia (Valerianaceae), together with the known one, nardostachin (3). The structures of compounds 1 and 2 were established on the basis of spectroscopic methods, including two dimensional (2D NMR) and high resolution fast atom bombardment mass spectrometry (HR-FAB-MS). PMID- 12736461 TI - Six new andrographolide metabolites in rats. AB - Six new andrographolide metabolites M-5-M-10 were isolated from rat urine, feces, and the contents of the small intestine. Three (M-5-M-7) as sulfate ester compounds were identified as new compounds. The structures of these six metabolites were determined to be 14-deoxy-12(R)-sulfo andrographolide 3-sulfate (M-5), 14-deoxy-12(S)-sulfo andrographolide 3-sulfate (M-6), 14-sulfo isoandrographolide 3-sulfate (M-7), 14-deoxy-11,12-didehydroandrographolide (M 8), isoandrographolide (M-9), and 14-deoxy andrographolide (M-10), respectively, based on chemical evidence and spectroscopic analysis. PMID- 12736462 TI - A novel ent-kaurane diterpenoid from the Croton tonkinensis GAGNEP. AB - A novel ent-kaurane diterpenoid, ent-1alpha-acetoxy-7beta,14alpha-dihydroxy-kaur 16-en-15-on has been isolated from leaves of Croton tonkinensis GAGNEP. Its structure was determined by a combination of spectroscopy, X-ray crystallographic analysis and the chemical reaction acetylation. PMID- 12736463 TI - Hemsleyatine, a novel C19-diterpenoid alkaloid with 8-amino group from Aconitum hemsleyanum. AB - The new compound hemsleyatine (1) was isolated along with four known C(19) diterpenoid alkaloids: indaconitine (4), yunaconitine (5), chasmanine (6), and talatisamine (7) from the roots of Aconitum hemsleyanum PRITZ. Structures were established by spectral analysis, including tow dimeusional (2D) NMR spectroscopy and a chemical method. Hemsleyatine (1) is the first C(19)-diterpenoid alkaloid bearing the 8-amino group. In addition, the assignments of some (13)C signals for pseudaconine (3) were revised by comparison with those of hemsleyatine (1). PMID- 12736464 TI - Separation of Leucas aspera, a medicinal plant of Bangladesh, guided by prostaglandin inhibitory and antioxidant activities. AB - According to the traditional usage of the plant for antiinflammation and analgesia, Leucas aspera was tested for its prostaglandin (PG) inhibitory and antioxidant activities. The extract showed both activities, i.e., inhibition at 3 x 10(-4) g/ml against PGE(1)- and PGE(2)-induced contractions in guinea pig ileum and a 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging effect. The separation guided by the activities in these dual assay methods provided eight lignans and four flavonoids, LA-1- -12, among which LA-1- -7 and LA-10- -12 were identified as nectandrin B, meso-dihydroguaiaretic acid, macelignan, acacetin, apigenin 7-O-[6"-O-(p-coumaroyl)-beta-D-glucoside], chrysoeriol, apigenin, erythro-2-(4-allyl-2, 6-dimethoxyphenoxy)-1-(4-hydroxy-3-methoxyphenyl)propan-1 ol, myristargenol B, and machilin C, respectively. LA-8 was determined to be (-) chicanine, the new antipode of the (+) compound, by spectroscopic methods including CD and ORD. Chiral-HPLC analysis of LA-9 showed that it was a mixture of two enantiomers, (7R, 8R)- and (7S, 8S)-licarin A. All of these components were first isolated from L. aspera. PG inhibition was observed in LA-1, LA-2, and LA-5, and antioxidant activity in LA-1- -3 and LA-8- -12. PMID- 12736465 TI - Chemical constituents of Taraxacum formosanum. AB - Three new compounds, taraxacine-A (1), taraxacine-B (2) and taraxafolin (3) together with twenty-five known compounds, which include two beta-carboline alkaloids, two indole alkaloids, two chlorophylls, two flavonoids, one coumarin, two triterpenoids, one monoterpenoid, one ionone, four steroids and eight benzenoids, were isolated and characterized from the fresh aerial parts of Taraxacum formosanum. Structures of new compounds were determined by spectral analysis. PMID- 12736466 TI - Dianion of sulfinylacetone as a synthetic equivalent of beta-enolate of propionic acid: a novel synthesis of carboxylic acids from alkyl halides with three-carbon elongation. AB - The reaction of the dianion of phenylsulfinylacetone with alkyl halides afforded beta-keto sulfoxides, which were first chlorinated with hexachloroethane and then treated successively with KH and t-BuLi to give carboxylic acids in three-steps in moderate overall yields from the alkyl halides. This procedure affords a good method for a synthesis of carboxylic acids from alkyl halides with three-carbon elongation. PMID- 12736467 TI - Inhibitory effect of TNF-alpha and IL-8 secretion by pimarane-type diterpenoids from Acanthopanax koreanum. AB - A new pimarane-type diterpene compound, acanthokoreoic acid A together with three known compounds, acanthoic acid, acanthol, and sumogaside were isolated from a CH(2)Cl(2) fraction of Acanthopanax koreanum by repeated column chromatography and reversed phase preparative HPLC. Acanthoic acid was isolated in high yields and showed potent inhibitory activity on the IL-8 secretion of the TNF-alpha stimulated human colon adenocarcinoma cell line HT-29 and on the TNF-alpha secretion of the trypsin-stimulated human leukemic mast cell line HMC-1. PMID- 12736468 TI - Synthesis of 6-substituted 9-benzyl-8-hydroxypurines with potential interferon inducing activity. AB - Various 6-substituted 9-benzyl-8-hydroxypurines were synthesized in order to investigate the structure-activity relationship at the 6-position of 9-benzyl-8 hydroxyadenine (1), which is a lead compound for the screening of interferon (IFN)-inducing activity. 6-Unsubstituted, mercapto-, methylthio- and hydroxy-9 benzyl-8-hydroxypurines (2-5) were prepared from 5-amino-1-benzyl-4-cyano-2 hydroxyimidazole (9). Synthesis of a 6-methoxy analog (6) was conducted from 5 amino-4-benzylamino-6-chloropyrimidine (13). 6-Alkylamino and acylaminopurines (7 and 8) were also prepared by alkylation and acylation of 1, respectively. Since these compounds (2-8) indicated no activity, it was found that a free amino group of 1 is required for the expression of IFN-inducing activity. PMID- 12736469 TI - Bahiensol, a new glycerolipid from a cultured myxomycete Didymium bahiense var. bahiense. AB - Bahiensol (1), a new glycerolipid with antimicrobial activity has been isolated from a cultured plasmodium of myxomycete Didymium bahiense var. bahiense and its planar structure was elucidated by spectral data. PMID- 12736470 TI - Water-soluble porphyrins appending platinum(II) complexes as binders for synthetic nucleic acid polymers. AB - Water-soluble porphyrins containing four platinum(II) complexes per molecule, [5alpha,10beta,15alpha,20beta-tetrakis(2-trans-(alpha,beta,alpha,beta-trans-Pt) and cis-(alpha,beta,alpha,beta-cis-Pt) [PtCl(NH(3))(2)]N-2 aminophenylporphyrin)], were synthesized and characterized. The binding of synthetic nucleotide polymers (poly(dG)-poly(dC), poly(dA)-poly(dT)) to the porphyrins was examined spectrophotometrically in aqueous solution. UV-vis spectral data suggested that these porphyrins bind to the nucleic acids by coordinative and Coulomb interactions. PMID- 12736471 TI - Comparison of outcomes for patients undergoing balloon angioplasty vs coronary stenting for acute myocardial infarction. AB - Randomized clinical trials have demonstrated that coronary stenting is more successful than balloon angioplasty in improving short- and long-term outcomes. However, it remains unknown whether those results can be generalized to broad based practice. This study aimed to determine whether the outcome for patients with acute myocardial infarction (AMI) undergoing coronary stent placement would be better than those undergoing balloon angioplasty. The risk-adjusted mortality and subsequent revascularization rates were compared for 2185 patients from a nationwide Japanese registry during 1997. A total of 1349 patients were treated with balloon angioplasty alone and 836 had stent placement. There were no statistically significant differences in the prevalence of demographic, clinical, and angiographic variables, except that the angioplasty group had a greater proportion of female patients and those with a left circumflex lesion. Unadjusted in-hospital mortality was comparable (7.6 vs 6.3%; p=0.28), despite higher angiographic success rate for the stent group (89.7 vs 97.7%; p<0.01). Adjusted odds ratio for in-hospital mortality was 0.75 (p=0.19). The same-admission bypass surgery rate was also similar. The 1.9-year post-discharge mortality rate was similar. The need for subsequent revascularization procedures was also similar, but restenosis was significantly lower in the stent group (34 vs 45%; p<0.01). The superiority of clinical outcome for stenting rather than balloon angioplasty could not be demonstrated in broad-based registry patients, despite technically successful results. PMID- 12736472 TI - Hemodynamic and hormonal effects of beraprost sodium, an orally active prostacyclin analogue, in patients with secondary precapillary pulmonary hypertension. AB - Earlier studies have shown that administration of beraprost sodium (BPS), an orally active prostacyclin analogue, improves hemodynamics in patients with primary pulmonary hypertension (PH), but it is not known whether BPS has beneficial effects in secondary precapillary PH. The present study investigated the hemodynamic and hormonal parameters of 18 patients with secondary precapillary PH (8 patients with chronic thromboembolic PH, 7 with collagen vascular disease, and 3 with residual PH after surgery for atrial septal defect). Hemodynamics were repeatedly measured by right heart catheterization. Treatment with BPS improved New York Heart Association (NYHA) functional class in 10 of the 18 patients and significantly decreased pulmonary vascular resistance by 17% (12.9+/-1.1 to 10.7+/-1.2 Wood units, p<0.01). Circulating brain natriuretic peptide and uric acid significantly decreased from 246+/-61 to 215+/-65 pg/ml and from 6.5+/-0.6 to 5.3+/-0.3 mg/dl, respectively. In summary, BPS therapy improved NYHA functional class, hemodynamics, and hormonal parameters in patients with secondary precapillary PH. Thus, oral administration of BPS may be a new therapeutic strategy for the treatment of secondary precapillary PH. PMID- 12736473 TI - Impact of myocardial angiotensin-converting enzyme activity on coronary vascular resistance and serum brain natriuretic peptide concentration in coronary bypass surgery. AB - Angiotensin-converting enzyme (ACE) inhibitors have cardioprotective effects in animals, but whether that occurs in humans is still controversial. The effect of myocardial ACE activity on coronary vascular resistance during coronary artery bypass surgery and on serum brain natriuretic peptide (BNP) concentration after surgery was studied in myocardial tissue sampled from the right atrium of patients during cardiac surgery (n=20). Tissue enzyme activity (nmol/min per mg protein) was measured using a photometric technique, and the flow rate and pressure upon antegrade infusion of a crystalloid cardioplegic solution was measured for calculating the coronary vascular resistance (mmHg. ml(-1). min( 1)). Serum BNP concentration (pg/ml) was measured on days 0 and 5 after the surgery. Linear regression between tissue ACE activity and coronary vascular resistance (y = 0.46x + 0.56, r=0.85) as well as serum BNP concentration on days 0 (y = 129x + 30, r=0.59) and 5 (y = 347x + 180, r=0.73) after the surgery was significant (x: ACE activity; y: coronary vascular resistance/serum BNP concentration). The results indicate that inhibition of myocardial ACE activity might improve coronary circulation during surgery and hence, cardiac function after surgery. PMID- 12736475 TI - Role of fibrillation cycle length in spontaneous and drug-induced termination of human atrial fibrillation. AB - The aim of this study was to investigate the mechanism of spontaneous termination of atrial fibrillation (AF) by comparing it with drug-induced termination on the basis of changes in fibrillation cycle length (FCL). Fast Fourier transform analysis was carried out on the electrocardiogram (ECG) records of 27 patients with paroxysmal AF without organic heart disease. In 8 patients with drug-induced termination of AF, spectral analysis of the data from surface ECG lead V1 was performed before and after class I antiarrhythmic drug infusion for 10 min. In 19 patients with spontaneous termination of AF, the analysis used the Holter ECG recordings at 10 min before the spontaneous termination and at the termination. FCL was calculated from the peak frequency of each epoch and the mean FCL and the coefficient of variation (CV) of FCL were determined from the data of 20 epochs. In the 8 episodes of drug-induced AF termination, the mean FCL increased significantly with class I drugs (from 151+/-17 to 203+/-21 ms, p<0.001), whereas in the 19 episodes of spontaneous termination, the mean FCL and CV of FCL at termination did not differ from those at 10 min before the termination. Of the 19 episodes, 10 episodes terminating in the morning showed a significant increase in the FCL of the last epoch of the termination period (from 158 +/-22 to 172+/-17 ms, p<0.05). In the 9 episodes terminating in either the afternoon or the evening, the FCL of the last epoch did not change significantly. Although drug induced termination of paroxysmal AF may depend on a gradual increase in FCL, the pattern of spontaneous termination may depend on the time of day. Spontaneous termination in the morning may be caused by an abrupt increase in FCL related to vagolytic autonomic balance. PMID- 12736474 TI - Effects of torasemide on left ventricular function and neurohumoral factors in patients with chronic heart failure. AB - The effect of torasemide and furosemide therapy was compared in 50 patients who had chronic heart failure and symptoms [NYHA class II-III] despite long-term therapy with both low-dose furosemide and angiotensin-converting enzyme inhibitors. In this randomized 6-month, open-label trial, baseline and follow-up echocardiograms and neurohumoral assays were obtained in 25 group F patients (continued same dose of oral furosemide at 20-40 mg/day) and in 25 group T patients (received torasemide at 4-8 mg/day in place of furosemide). At 6 months, parameters were unchanged in group F whereas the group T patients had a lower left ventricular end-diastolic diameter (p<0.005) and left ventricular mass index (p<0.005) with improved Doppler filling parameters, decreased plasma B-type natriuretic concentration (p<0.001) and increased plasma concentrations of active renin (p<0.005) and aldosterone (p<0.001). The magnitude of these changes appeared dose dependent and it is suggested these favorable effects of switching from furosemide to torasemide may be related to aldosterone receptor blockade. PMID- 12736476 TI - Greater impairment of right ventricular systolic function in patients with anterior myocardial infarction because of the extent of proximal lesions. AB - The present study examined the influence of the extent of the ischemic area on right ventricular (RV) systolic function and the relation between the RV global and regional systolic function in patients with anteroseptal myocardial infarction (MI). Biplane right ventriculography was performed in 15 subjects as the control group, and 46 patients with anteroseptal MI as the MI group. Three dimensions of the RV (the long axis dimension [LA], the anterior-posterior dimension [AP] and the septum-free wall dimension [SF]) were examined to assess regional function The MI group had a larger right ventricular end-systolic volume index and lower right ventricular ejection fraction than the control group. The more proximal the coronary lesion, the lower was the ejection fraction of the RV in the MI group. The MI group had lower percent shortening (% shortening) of the SF than the control group, but there were no significant change in the % shortening of AP and LA between the groups. The results suggest that the degree of impairment of RV systolic function depends on the extent of the infarcted area, and that the impairment is mainly from a reduction in the %shortening of the SF. PMID- 12736478 TI - Observation of the ischemic cascade in humans using contrast echocardiography during dobutamine stress. AB - Experimental studies have postulated the ischemic cascade and the present study was designed to elucidate whether it can be observed in the clinical setting. Fifty-three patients suspected of having coronary artery disease were studied. Myocardial perfusion abnormalities (MPA) and wall motion abnormalities (WMA) were assessed simultaneously by infusion of Levovist during dobutamine stress echocardiography. Time - intensity data of myocardial opacification were fitted for Y=A (1-e(-)(beta) (t)) from which the rate of increase (beta) of intensity were derived both at rest and during stress. Wall motion was also given a score. Bright opacification was observed in 50 patients: 25 showed significant stenosis (>50%) in the left anterior descending artery (group II) on coronary angiography and 25 did not (group I). Significant differences were found in the beta ratio (stress/rest) between the 2 groups at a low-dose (2.0+/-0.3 vs 1.5+/-0.5, p<0.05) and at a high-dose of dobutamine (2.7+/-1.0 vs 1.1+/-0.5, p<0.001), whereas the wall motion score differed only at a high-dose. Of the 25 patients in group II, MPA preceded WMA in 12, both occurred at the same stage in 12, and neither MPA nor WMA was seen in 1. These data prove the ischemic cascade clinically, using contrast echocardiography, by demonstrating that MPA precede WMA during dobutamine stress in patients with coronary stenosis. PMID- 12736479 TI - Use of exercise cardiac rehabilitation after acute myocardial infarction. AB - The purpose of this study was to determine the rate of participation of patients after acute myocardial infarction (AMI) in phase II cardiac rehabilitation with exercise training (ie, exercise cardiac rehabilitation, ECR) in Japan. Forty-six hospitals treating patients with AMI were surveyed for their implementation of phase II ECR after AMI in 1996-98. Of the 46 hospitals, 19 were approved and 27 were not approved for health insurance payment for ECR. A total of 13685 patients with AMI were admitted to the 46 hospitals. There were no differences between approved and non-approved hospitals in the annual number of patients with AMI (Approved, 117+61 vs Non-approved, 86+71 patients per hospital, NS), the rate of performance of emergency coronary angioplasty (63+16 vs 65+20%, NS), or the rate of emergency coronary stenting (31+16 vs 34+22%, NS). However, ECR was performed routinely in 84.2% (16/19 hospitals) of the approved hospitals, but in only 22.2% (6/27 hospitals) of the non-approved hospitals (p<0.001). Although the participation rate of AMI patients in ECR was 21.0% (2875/13685 patients) overall, it was markedly lower in the non-approved hospitals (8.0%, 557/6999 patients) than in the approved hospitals (34.7%, 2318/6686 patients, p<0.0001). Based on the present result, the overall rate of participation of AMI patients in ECR in Japan was estimated at 4.8-11.7%. Despite similar patient volumes and acute phase interventional treatment of AMI between the hospitals approved and not approved for health insurance payment for ECR, ECR was markedly underused in the non-approved hospitals in Japan. To promote ECR for all AMI patients in Japan, the number of hospitals approved for ECR should be substantially increased. PMID- 12736477 TI - Noninvasive assessment of coronary artery disease by multislice spiral computed tomography using a new retrospectively ECG-gated image reconstruction technique. AB - The present study was designed to investigate the accuracy of multislice spiral computed tomography (MSCT) in detecting coronary artery disease, compared with coronary angiography (CAG), using a new retrospectively ECG-gated reconstruction method that reduced cardiac motion artifact. The study group comprised 54 consecutive patients undergoing MSCT and CAG. MSCT was performed using a SOMATOM Volume Zoom (4-detector-row, Siemens, Germany) with slice thickness 1.0 mm, pitch 1.5 (table feed: 1.5 mm per rotation) and gantry rotation time 500 ms. Metoprolol (20-60 mg) was administered orally prior to MSCT imaging. ECG-gated image reconstruction was performed with the reconstruction window (250 ms) positioned immediately before atrial contraction in order to reduce the cardiac motion artifact caused by the abrupt diastolic ventricular movement occurring during the rapid filling and atrial contraction periods. Following inspection of the volume rendering images, multiplanar reconstruction images and axial images of the left main coronary artery (LMCA), left anterior descending artery (LAD), left circumflex artery (LCx) and right coronary artery (RCA) were obtained and evaluated for luminal narrowing. The results were compared with those obtained by CAG. Of 216 coronary arteries, 206 (95.4%) were assessable; 10 arteries were excluded from the analysis because of severe calcification (n=4), stents (n=3) or insufficient contrast enhancement (n=3). The sensitivity to detect coronary stenoses >or=50% was 93.5% and the specificity to define luminal narrowing <50% was 97.2%. The positive predictive value and the negative predictive value were 93.5% and 97.2%, respectively. The sensitivity was still satisfactory (80.6%) even when non-assessable arteries were included in the analysis. The new retrospectively ECG-gated reconstruction method for MSCT has excellent diagnostic accuracy in detecting significant coronary artery stenoses. PMID- 12736481 TI - Impact of diabetes mellitus on angiographically silent coronary atherosclerosis. AB - Constrictive remodeling occurs in significant atherosclerotic lesions of the diabetic patient, but the impact of diabetes mellitus (DM) on the angiographically normal coronary artery is still unclear. Morphometric analysis using intravascular ultrasound (IVUS) prior to intervention evaluated 54 sites in 33 DM patients and 106 in 62 non-diabetic patients. Vessel area (VA) and lumen area (LA) were measured at angiographically normal sites in the vessel. Plaque area (PA) was calculated as VA - LA. Percentage plaque area (%PA) was calculated as PA VA. Even in the angiographically normal site, mild coronary atherosclerosis was detected by IVUS in both groups. In the patients with DM, VA and LA were significantly smaller than in the non-diabetic patient (15.5 vs 17.8 mm(2), p<0.01; and 10.1 vs 12.2 mm(2), p<0.01 respectively), whereas % PA was similar (34.5 vs 31.6%). At angiographically normal sites where mild coronary atherosclerosis is detected by IVUS, the coronary artery of diabetic patients is smaller than that of the non-diabetic. These results suggest impaired compensatory enlargement or some other constrictive mechanism has already occurred in the early stages of coronary atherosclerosis in patients with DM. PMID- 12736480 TI - Novel approach to the quantitation of regional left ventricular systolic and diastolic function using tissue Doppler imaging to create a myocardial velocity profile and gradient. AB - The myocardial velocity profile (MVP) and gradient (MVG) between the endocardium and epicardium of the left ventricular (LV) wall measured by color-coded tissue Doppler imaging (TDI) are new indices for evaluating regional LV myocardial function. However, accurate recording and measurement of the MVP is difficult using conventional methodology because of the stochastic nature of the ultrasound signal; that is, the effect of speckled noise. The aim of this study was to validate the accuracy and establish the validity of a newly developed method for measuring the MVP and MVG using 10 clinically normal controls and 10 patients with a hypertensive hypertrophied LV posterior wall. A non-isotropic, averaging algorithm was developed that was capable of obtaining a stable MVP (averaged MVP). Averaged MVP was recorded using parasternal, LV short-axis, color-coded TDI, placing regions of interest along the LV posterior wall with the reference point for angle-correction being at the center of LV contraction. The velocity from epicardium to endocardium within the region of interest was automatically angle-corrected to calculate the velocity component radially relative to the LV cavity and was spatially averaged along the circumference within the region of interest. Inter- and intraobserber variabilities of measurements were lower in the averaged MVP and MVG than in the conventional MVP and MVG. The correlation coefficients of the linear regression lines of systolic and early diastolic MVPs in the LV posterior wall were higher in all controls and hypertensive patients with the averaged method than with the conventional TDI procedures. The mean peak systolic and early diastolic MVGs were lower in the hypertensive group than in the controls. In conclusion, the newly developed averaged MVP provides a stable and reproducible index for the quantitative assessment of regional LV myocardial function. PMID- 12736482 TI - Detection of coronary artery aneurysms, stenoses and occlusions by multislice spiral computed tomography in adolescents with kawasaki disease. AB - In patients with Kawasaki disease (KD), serial evaluation of coronary artery aneurysms (CAAs) and luminal narrowing is essential for risk stratification and therapeutic management. Therefore, non-invasive assessment of the status of the coronary artery is of utmost importance in patient management. Multislice spiral computed tomography (MSCT) permits non-invasive visualization of the entire coronary artery system and was used in the evaluation of 4 patients with KD. CAAs and high-grade coronary artery stenoses were detected by MSCT and corroborated the findings of coronary angiograms performed within the previous 2 years. MSCT has the potential to be the standard diagnostic tool in adolescents with KD. PMID- 12736483 TI - Characteristics and 1-year prognosis of medically treated patients with chronic heart failure in Japan. AB - The study was designed to characterize patients with chronic heart failure (CHF) in Japan in terms of the etiologies and prognosis. CHF was defined by ejection fraction (EF >or=50%), left ventricular diastolic dimension (LVDD >or=55 mm) or a past history of congestive heart failure. Among the 721 recruited patients, the most frequent etiology for CHF was dilated cardiomyopathy (DCM) in patients aged less than 59 years, and valvular heart disease (VHD) in those aged 70 years or more. The 1-year crude mortality was 8% overall and 12% in patients with myocardial infarction (MI). Sudden death accounted for 40% of the total deaths among all patients, and 60% in patients with MI. Multivariate logistic regression analysis showed that brain natriuretic peptide (BNP) was a consistent prognostic marker in CHF patients with a variety of etiologies. Total death and hospitalization because of heart failure were significantly less frequent in patients with BNP less than 100 pg/ml. In conclusion, the etiologies of Japanese CHF appear to be more diverse than those of other Western countries, but BNP is an excellent prognostic marker despite the etiological diversity. Sudden, unexpected death in CHF patients is also a serious problem in Japan. A nation wide epidemiologic study should be done to characterize Japanese CHF. PMID- 12736484 TI - Effects of flecainide on the electrophysiological properties of atrial vulnerability in humans. AB - The aims of this study were to evaluate the changes in the electrophysiological characteristics of the right atrium after the administration of flecainide and to clarify whether flecainide has a selective effect on human atrial tissue. Electrophysiological measurements were made in 38 patients, before and after intravenous administration of flecainide (2 mg/kg per 10 min). The effective refractory period of the right atrium (ERP-A), maximum conduction delay (Max.CD), repetitive atrial firing zone (RAFZ), fragmented atrial activity zone (FAAZ), and conduction delay zone (CDZ) were studied in the patients who were divided into 2 groups based on whether repetitive atrial firing (RAF) was induced in the baseline study. Flecainide significantly prolonged the ERP-A (202+/-22 to 238+/ 33 ms, p<0.001) and shortened Max.CD (77+/-17 to 63+/-32 ms, p<0.05) in the patients with RAF, but not in those without RAF in the baseline study. After flecainide administration, there were significant reductions in the RAFZ (43+/-22 to 13+/-19 ms, p<0.0001), FAAZ (51+/-22 to 28+/-26 ms, p<0.001) and CDZ (70+/-21 to 48+/-30 ms, p<0.01) in the patients with RAF. However, atrial fibrillation (AF) was induced by stimulation after flecainide in 2 patients without RAF in the baseline study. There was a significant negative correlation between the ERP-A in the baseline study and the change in the ERP-A upon flecainide administration (r=0.45, p<0.01). Flecainide may preferentially activate the substrate for AF and RAF, but that action is mainly based on the electrophysiological characteristics found in the baseline study. PMID- 12736485 TI - Administration of atrial natriuretic peptide attenuates reperfusion phenomena and preserves left ventricular regional wall motion after direct coronary angioplasty for acute myocardial infarction. AB - To evaluate the effects of synthetic human atrial natriuretic peptide (hANP) on myocardial reperfusion injury and left ventricular remodeling, 19 patients within 12 h of a first attack of anterior myocardial infarction (AMI) underwent intracoronary injection of 25 microg of hANP immediately after coronary angioplasty, combined with intravenous infusion of 0.025 microg x kg(-1) x min( 1) of hANP initiated on admission for 1 week (hANP group); 18 similar patients had saline administered (control group). The incidences of premature ventricular contraction, ventricular tachycardia and/or fibrillation in the hANP group were significantly less than in the control group after coronary angioplasty. Left ventricular ejection fraction was significantly greater and left ventricular end diastolic volume index was significantly smaller 6 months after coronary angioplasty. Left ventricular regional wall motion of the infarcted segments significantly increased. Thus, hANP remarkably suppressed reperfusion phenomena and preserved left ventricular function through improvement of regional wall motion of the infarcted segments after coronary angioplasty. PMID- 12736487 TI - Radiofrequency catheter ablation of various kinds of arrhythmias guided by virtual electrograms using a noncontact, computerized mapping system. AB - Three-dimensional visualization of cardiac activation has become important for providing further insights into the pathophysiological mechanisms of arrhythmias and to increase the efficacy of catheter ablation. The noncontact mapping system enables a single-beat analysis of the reconstructed geometry of the cardiac chamber. In 8 patients with various kinds of arrhythmias (3 with atrial flutter, 2 with right ventricular outflow tract ventricular tachycardia, 1 with idiopathic left ventricular tachycardia, 1 with atrioventricular nodal reentrant tachycardia and 1 with concealed Wolff-Perkinson-White syndrome), non-contact mapping using an EnSite 3000 system was performed for the guidance of catheter ablation. The optimal sites for successful ablation were detected and all of these arrhythmias were successfully eliminated with the radiofrequency energy applications without any adverse effects. The computerized EnSite 3000 mapping system described here computes accurate isopotential maps that are a useful guide for catheter ablation. PMID- 12736486 TI - Transmural heterogeneity of the action potential configuration in the feline left ventricle. AB - There are M cells in the canine, rabbit, guinea pig, and human left ventricle (LV), but it is not known if they are present in the feline LV. Arterially perfused feline LV preparations were used for the recording of transmembrane action potentials from the epicardium (Epi), midmyocardium (M) and endomyocardium (Endo) under control conditions (n=12) and in the presence of I(Ks) blocker (chromanol 293B: 10 micromol/L, n=6) or I(Kr) blocker (E-4031: 2 micromol/L, n=6). The steady-state action potential duration at 90% repolarization and cycle length (APD90/CL) relation was obtained and fitted by the hyperbolic function APD(90) = CL/[(a x CL) + b]. In control, the shortest and longest action potential duration (APD) were observed in Epi and M, respectively, and the APD(90)/CL-relation curve was steeper in the M or Endo than in the Epi. Chromanol 293B prolonged APD in Epi, but not in M or Endo, resulting in no significant difference of the APD(90)/CL-relation curve among the 3 regions. E-4031 markedly, but homogeneously, prolonged APD in all regions, giving rise to decreased transmural dispersion of repolarization. In conclusion, there exists an M cell layer with a longer APD than the Epi and Endo layers and there is transmural electrical heterogeneity in the feline LV; however, the response to I(Kr) blocker is different from that of the canine LV probably because of species differences in the I(Kr) and I(Ks). PMID- 12736488 TI - Spontaneous rupture of the ascending aorta: case report and review. AB - Spontaneous nontraumatic rupture of the ascending aorta occurred in a hypertensive patient. The clinical findings suggested acute aortic dissection, and echocardiography showed a large pericardial effusion. Computed tomography scanning did not indicate aortic dissection, but aortography in 3 projections revealed an area of intimal disruption similar to the niche of an ulcer. The patient underwent replacement of the ascending aorta and proximal aortic arch, and the postoperative course was uneventful. PMID- 12736489 TI - Simultaneous coronary - subclavian and vertebral - subclavian steal syndrome. AB - Atherosclerotic disease of the coronary artery may simultaneously involve the subclavian artery, and a significant stenosis of the left subclavian artery may result in recurrent myocardial ischemia in patients with patent left internal mammary artery (LIMA) grafts because of 'coronary steal' through the LIMA. Isometric exercise of the left arm may improve myocardial perfusion through vertebral - subclavian steal by flow reversal in the ipsilateral vertebral artery because of the change in the pressure gradient between the circle of Willis and the distal subclavian artery. The present patient had coronary steal through a LIMA after coronary artery bypass surgery and a transient vertebral - subclavian steal with improved myocardial perfusion as a result of exercise of the left arm. PMID- 12736490 TI - Vasospastic angina in a 16-year-old female. AB - A 16-year-old female with a family history of coronary artery disease was referred to hospital because of recurrent chest oppression unrelated to exertion. Although a selective coronary angiogram showed no atherosclerotic lesions, coronary spasm was provoked by acetylcholine in the left coronary artery, accompanied by chest pain and depression of blood pressure. This is the youngest healthy young female to be diagnosed with coronary vasospasm by provocation test. Because there are no risk factors, the vasospasm must be related to unknown hereditary factors. PMID- 12736491 TI - Ruptured aneurysm of the sinus of valsalva coexisting with a ventricular septal defect and single coronary artery. AB - A 26-year-old man had been diagnosed with a cardiac murmur from birth. In 1998, he was admitted to hospital because of slight fatigue. A grade 5/6 continuous murmur was audible near the right sternal border at the second intercostal space. Doppler echocardiography detected an abnormal flow that suggested that an aneurysm of the right coronary sinus of Valsalva had ruptured into the right ventricular inflow tract. Blood tests showed a 19% step-up in oxygen saturation value between the right atrium and right ventricle, indicating a ventricular septal defect with left to right shunt. Coronary angiography revealed a single coronary artery. Surgical repair was carried out and the patient made an uneventful recovery. This rare combination of a ruptured aneurysm of the sinus of Valsalva coexisting with a ventricular septal defect and a single coronary artery has not been reported previously. PMID- 12736492 TI - Assessment of radioactive residues arising from radiolabel instability in a multiple dose tissue distribution study in rats. AB - Our study objectives were: To quantitatively determine the effect of radiolabel instability on terminal phase radioactive tissue residues in a multiple dose tissue distribution study. To quantitatively compare tissue residue artifacts (non drug-related radioactivity) from two chemically-distinct radiolabel locations. To conduct a definitive multiple dose tissue distribution study using the better of the two radiolabeled compounds. We compared the excretion and tissue distribution in rats of [(14)C]linezolid, radiolabeled in two different locations, after 7 consecutive once daily [(14)C] oral doses. The radiolabels were in the acetamide (two carbon) and oxazolidinone (isolated carbon) functional groups. Terminal phase tissue residue and excretion data were compared to data from rats dosed orally with [(14)C]sodium acetate. Drug-related radioactivity was excreted rapidly over 24 h. After a single dose, the acetamide and oxazolidinone radiolabel sites both gave 3% of dose as exhaled (14)CO(2). After 7 daily [(14)C] oral doses, terminal phase radioactive tissue residues were higher from the acetamide radiolabel, relative to the oxazolidinone radiolabel, and were primarily not drug-related. In the definitive tissue distribution study, low concentrations of drug-related radioactivity in skin and thyroid were observed. We conclude that although small amounts of radiolabel instability do not significantly affect single dose tissue radioactivity C(max) and AUC, artifacts arising from radiolabel instability can prolong the apparent terminal phase half life and complicate study data interpretation. When possible, it is always preferable to use a completely stable radiolabel site. PMID- 12736493 TI - Subcellular distribution of mouse mevalonate pyrophosphate decarboxylase. AB - Mevalonate pyrophosphate decarboxylase (MPD) is considered to be a cytosolic protein. Recently, other groups reported that MPD is mostly located in the peroxisomes. In this study, we examined whether the expression of MPD in mice depends on the proliferation of peroxisomes, and whether MPD is predominantly located in the peroxisomes or the cytosol of mice. No increase in the protein level of MPD was observed in the crude extract of the livers of mice administered with peroxisome proliferative drugs. The result suggests that the expression of MPD is independent of the proliferation of peroxisomes, and may be maintained via a specific regulatory mechanism, different from the regulation of the expression of peroxisome proliferator-activated receptor alpha. When the subcellular distribution of MPD in mouse melanoma (B16F10) cells was examined by cell fractionation, MPD was detected in the cytosol of B16F10 cells, but not in the peroxisomes. In permeabilized B16F10 cells treated with digitonin, which lack cytosolic enzymes, 80% and 20% of MPD, 75% and 25% of lactate dehydrogenase, or 2% and 98% of catalase, existed in the medium and in the cell, respectively. From these results, it indicated that MPD was predominantly located in the cytosol and did not exist in the peroxisomes of B16F10 cells. PMID- 12736495 TI - Dependence of hydrolysis of beta-lactams with a zinc(II)-beta-lactamase produced from Serratia marcescens (IMP-1) on pH and concentration of zinc(II) ion: dissociation of Zn(II) from IMP-1 in acidic medium. AB - The pH dependence for the hydrolysis of beta-lactam antibiotics by a metallo-beta lactamase (IMP-1) produced from Serratia marcescens was investigated varying the concentration of Zn(II). The activity of IMP-1 for imipenem was decreased at pH less than pH 5.3 without external addition of Zn(II) ions but was recovered with addition of Zn(II). Varying the concentration of external Zn(II), the molar activity of the enzyme, k(obs), that was defined by the velocity of hydrolysis of imipenem/concentration of IMP-1 was expressed by k(obs)=v(init)/[E](T)=k(max)[Zn]/(K(d)+[Zn]) in which K(d) stands for the dissociation constant between Zn(II) and IMP-1. The dissociation constants, K(d), vary with pH; K(d)=840 x 10(-6) M at pH 4.3 and K(d)=0.19 x 10(-6) M at pH 6.0. The plot of -log K(d) against pH showed a straight line having a slope of 4.0 below pH 5.0, showing the existence of four functional groups which may be protonated upon dissociation of Zn(II) ion(s). The k(cat), K(m), and k(cat)/K(m) of hydrolysis of imipenem and cephalothin in the presence of sufficient concentration of Zn(NO(3))(2) for saturation of IMP-1 with Zn(II) showed similar dependency to each other on pH between pH 6.0 and 9.0. PMID- 12736494 TI - Diagnostic significance and clinical applications of chimeric genes in Ewing's sarcoma. AB - Ewing's sarcoma (ES) is one of the most malignant bone and soft tissue tumors in childhood. Morphologically, ES belongs to the small round cell tumors (SRCT). ES, peripheral primitive neuroectodermal tumor (PNET), and Askin's tumor are classified as ES family tumors (ESFT) because they share a common chromosomal translocation. The EWS-FLI1 chimeric gene is generated by t (11; 22). Other reciprocal translocations resulting in formation of chimeric genes between EWS and ETS family genes (ERG, ETV1, E1AF, and FEV) are t (21; 22), t (7; 22), t (17; 22), and t (2; 22), respectively. Although it is generally difficult to distinguish ES from SRCT, we could easily and quickly distinguish ES from other SRCT by using reverse transcription polymerase chain reaction (RT-PCR). We looked for specific chimeric genes in 23 tumor samples, including three ES clinical samples. We detected five chimeric genes in the three ES samples. Three chimeric genes, all EWS-FLI1, were detected in one ES sample. Different chimeric genes, EWS-ERG and EWS-ETV1, were detected in the other two ES samples. Moreover, because we could not detect specific chimeric genes in samples from non-ESFT, it may be possible to use this technique to diagnose ESFT and to detect tumor cell contamination before hematopoietic stem cell transplantation. PMID- 12736496 TI - Antioxidant effects in the quinone fraction from Auxemma oncocalyx TAUB. AB - In previous studies in vitro we showed that the quinone fraction (QF) from the heartwood of Auxemma oncocalyx TAUB. presented antiplatelet and antioxidant activities. In the present work, the QF antioxidant property was evaluated in models of CCl(4)-induced hepatotoxicity in rats, and prolongation of pentobarbital-induced sleeping time in mice. Our results showed that levels of plasma glutamate-pyruvate-transaminase (GPT), as well as glutamate-oxalate transaminase (GOT), were increased by the administration of CCl(4). On the other hand, only GPT levels were reduced by the QF treatment. Pentobarbital sleeping time was prolonged by the administration of CCl(4) and reduced by the QF treatment. Moreover, QF did not alter the pentobarbital-induced sleeping time. In conclusion, we showed that QF, represented mainly by oncocalyxone A, has hepatoprotective activity, and this effect is at least in part due to the antioxidant activity of this quinone. PMID- 12736497 TI - The metabolic stability of acyl-CoA: cholesterol O-acyltransferase (ACAT) inhibitors, N-(4-benzyloxy-3, 5-dimethoxycinnamoyl)-N'-(2, 4 dimethylphenyl)piperazine (YIC-708-424) and its derivatives in rat liver and intestinal epithelium. AB - The metabolic stability of the acyl-CoA: cholesterol O-acyltransferase (ACAT) inhibitor N-(4-benzyloxy-3, 5-dimethoxycinnamoyl)-N'-(2, 4 dimethylphenyl)piperazine (YIC-708-424) and its n-alkoxy derivatives containing an alkyl chain of 3 or 7 to 10 carbons, which exhibited different hypocholesterolemic activities, was investigated in vivo and in vitro in rats. After the oral administration of YIC-708-424 to rats at a dose of 5 mg/kg/d for 7 d, the parent compound was not detected in the blood. On the other hand, when the n-alkoxy derivatives were administered to rats, an increase in the alkyl chain length produced a progressive increase in the blood concentration of the parent compound. Both in the blood of rats administered YIC-708-424 and in the reaction mixture after the incubation of YIC-708-424 with rat hepatic 9000 x g supernatants, an inactive major metabolite, N-(4-benzyloxy-3, 5 dimethoxycinnamoyl)-N'-(4-carboxyl-2-methylphenyl)piperazine, was observed. The ratio of the maximum velocity to the apparent Michaelis-Menten constant (V(max)/K(m)) for the degradation of the n-propyloxy derivative in rat hepatic and intestinal microsomes was almost equivalent to that of YIC-708-424. On the other hand, an increase in the alkyl chain length of n-alkoxy derivatives produced a progressive decrease in V(max)/K(m) for the degradation of these compounds. Additionally, the in vivo hypocholesterolemic activities of YIC-708 424 and its n-alkoxy derivatives were positively correlated with the blood concentration of the parent compound and were negatively correlated with their V(max)/K(m). These results suggest that the metabolic stability of ACAT inhibitors in the liver and intestinal epithelium, which are the major target organs of these compounds, has a strong influence on their pharmacological activities in vivo. PMID- 12736499 TI - Pharmacokinetics of rhein from Onpi-to, an Oriental herbal medicine, in rats. AB - Onpi-to, an herbal medicine composed of five crude drugs (Rhei Rhizoma, Glycyrrhizae Radix, Ginseng Radix, Zingiberis Rhizoma and Aconiti Tuber), was administered orally to rats. Onpi-to includes 1.240% of total potential rhein derived from sennoside A, sennoside B, rhein 8-O-glucopyranoside and rhein. Plasma, urinary and biliary levels of rhein were determined by an HPLC-UV method. The plasma levels displayed curves characterized by maximum peaks at 8.3+/-5.2 min, 8.3+/-5.2 min and 20.0+/-21.9 min following dosages of 125, 250 and 500 mg/kg with mean concentrations of 1302.5+/-926.4, 2973.6+/-684.3 and 3118.8+/ 1701.2 ng/ml, respectively, followed by a subsequent decline. Area under the concentration-time curve (AUC)(0-48 h) at doses of 125, 250 and 500 mg/kg were 752.3+/-321.5, 2443.3+/-554.4 and 4443.2+/-2641.3 ng.h/ml, respectively. In female rats, rhein plasma levels showed curves which had a maximum peak at 45.0+/ 16.4 min after a dosage of 250 mg/kg with mean concentration of 3058.0+/-1533.7 ng/ml, followed by a subsequent decline. AUC(0-48 h) was 5537.7+/-1876.0 ng.h/ml. The cumulative urinary excretion of rhein and of conjugated rhein was 3.14+/ 1.56% and 38.21+/-18.87% of dose, respectively, 48 h after dosing at 500 mg/kg of Onpi-to in male rats. The cumulative biliary excretion of rhein was 1.34+/-0.44% of dose 48 h after dosing at 500 mg/kg of Onpi-to in male rats. PMID- 12736498 TI - Pharmacokinetics of (-)-epicatechin-3-O-gallate, an active component of Onpi-to, in rats. AB - (-)-Epicatechin-3-O-gallate (ECG), a component of Rhei Rhizoma, is one of the active components of Onpi-to, a herbal medicine composed of five crude drugs (Rhei Rhizome, Glycyrrhizae Radix, Ginseng Radix, Zingiberis Rhizoma and Aconiti Tuber), which has been used in patients with chronic renal failure. Pharmacokinetics of ECG was investigated in male rats employing an HPLC electrochemical detection method. 1. Following oral administration of ECG, ECG plasma levels revealed curves characterized by peaks at 0.065, 0.063 and 0.085 h corresponding to dosages of 12.5, 25.0 and 50.0 mg/kg at mean concentrations of 49.62, 212.89 and 464.04 ng/ml, respectively. Plasma levels subsequently declined bi-exponentially. ECG demonstrated nonlinear pharmacokinetics in terms of C(max) and AUC(0-inf). 2. The absolute bioavailability values (F) were 1.02, 1.47 and 3.30% at doses of 12.5, 25.0, and 50.0 mg/kg, respectively. 3. Following intravenous injection of ECG, plasma levels of ECG decreased with the gamma elimination half-life (t(1/2gamma)) of 4.03 h. 4. Following oral administration of ECG, urinary levels of ECG were lower than the quantitation limit. Moreover, cumulative excretion of the metabolites, delta-(3,4-dihydroxyphenyl)-gamma valerolactone and delta-(3-methoxy-4-hydroxyphenyl)-gamma-valerolactone, was 2.45 and 0.23% of dose, respectively, up to 30 h after dosing. PMID- 12736500 TI - Effects of prednisolone on the cutaneous reaction and skin barrier function in mice treated with a hapten. AB - Glucocorticoids are effective drugs for the treatment of allergic skin diseases. In the present study, we observed the effects of prednisolone on the cutaneous reaction and skin barrier function in mice treated with a hapten, 2,4 dinitrofluorobenzene. Repeated hapten application onto the mouse ear resulted in a potent ear swelling with an elevation of specific serum IgE. The ear swelling appeared following the second application of the hapten and peaked at 24 h after each application. Specific serum IgE was detected first after the fourth hapten application. Topical treatment with prednisolone apparently suppressed the swelling, whereas it failed to affect the serum specific IgE level. The hapten application caused an increase in transepidermal water loss, which was potently inhibited by prednisolone, although the water content was not affected. Amounts of triglyceride and cholesterol in the ear skin increased after repeated hapten applications, whereas the relative amount of free fatty acid and ceramide diminished. Prednisolone exhibited an inhibitory effect on the changes in lipid content. Thus prednisolone apparently inhibits the alteration of skin barrier function caused by hapten application as well as the cutaneous reaction. PMID- 12736501 TI - The effect of chondroitin sulfate against CCl4-induced hepatotoxicity. AB - This study was conducted to develop a new biomaterial to be used for an antioxidative drug. In this study, the hepatoprotective effect of chondroitin sulfate (CS) (100 mg/kg and 200 mg/kg body weight) was investigated at the antioxidative enzyme levels of liver total homogenate and mitochondria fraction. And the carbone tetrachloride (CCl(4))-induced rats were used as hepatotoxic models. The CCl(4) induced rat has been widely used as a hepatotoxic model due to its practicality, convenience and cost effectiveness since the generation of free oxygen radicals by CCl(4) injection was proposed as an important causative agent of hepatotoxicity. Malondialdehyde (MDA) levels were determined as well as the activities of superoxide dismutase (SOD), catalase (CAT), reduced-glutathione (GSH), oxidized-glutathione (GSSG) and glutathione peroxidase (GPx) in the liver. In addition, histopathology of liver tissue was investigated. Liver antioxidative enzyme activity was elevated while MDA concentration was decreased in all CS treated animals. The results demonstrated that CS protected oxidative stress in a dose dependent manner. Moreover, inflammation and cirrhosis in liver tissue of CS treated group were significantly decreased. It gave us an impression that CS might be a radical scavenger. PMID- 12736502 TI - Flow cytometric estimation of cytotoxic activity of rhodexin A isolated from Rhodea japonica in human leukemia K562 cells. AB - We have examined the cytotoxic effect of rhodexin A isolated from the extract of Rhodea japonica on human leukemia K562 cells using a flow cytometer and compared it with that of ouabain. Rhodexin A at 30 nM started to attenuate growth without affecting viability and further increases in the concentration of rhodexin A (100 nM or more) completely inhibited growth with decreasing viability. Rhodexin A at 30-100 nM increased the G(2)M population, but decreased the G(0)G(1) population, suggesting cell cycle arrest in the G(2)M phase. Rhodexin A at 100 nM increased the number of cells with hypodiploid DNA, indicating that rhodexin A induced apoptosis. The potency of rhodexin A to inhibit growth was greater than that of ouabain. The results indicate that rhodexin A exerts a potent inhibitory action on the growth of human leukemia K562 cells by inducing cell cycle arrest and apoptosis. Rhodexin A may also be a candidate for cancer treatment because there have been clinical reports of tumor regression in patients taking cardiac glycosides. PMID- 12736503 TI - Cytotoxicity of some azines of acetophenone derived mono-Mannich bases against Jurkat cells. AB - Acetophenone derived mono-Mannich bases (Ig1-Ig4), 1-aryl-3-amino-1-propanone hydrochlorides, which are known to have cytotoxicity in Jurkat cells, were synthesized. Then, they were converted to corresponding azine derivatives (D1 D4), N, N'-bis(3-amino-1-aryl-propylidene)hydrazine dihydrochlorides, which are bifunctional agents. The aryl part was replaced by phenyl in Ig1, Ig2, Ig3, D1, D2, and D3, and by p-hydroxyphenyl in Ig4 and D4. The amine part was replaced by dimethylamine in Ig1, D1, Ig4 and D4, by piperidine in Ig2 and D2, and by morpholine in Ig3 and D3. The aim of this study was to investigate whether the modification in chemical structure, converting the mono-Mannich base to a corresponding azine derivative, improves the cytotoxicity. In addition, the effect of the representative compound, D3, N, N'-bis(3-morpholine-4-yl-1 phenylpropylidene)hydrazine dihydrochloride, on cellular glutathione level after 1 h exposure in phosphate buffer at 37 degrees C was also determined to provide information on a possible mechanism of cytotoxic action. Compounds D2-D4 are reported for the first time in this study. Except for Ig2 and D2, the cytotoxicity of mono-Mannich bases, Ig1, Ig3 and Ig4 and corresponding azine derivatives, D1, D3 and D4 were higher than the reference compound 5-FU. Azine derivatives D1 and D4 had almost equal cytotoxic potency with corresponding mono Mannich bases Ig1 and Ig4, respectively. On the other hand, azine derivatives D2 and D3, had 1.28 and 1.90-times less cytotoxicity in Jurkat cells compared with the mono-Mannich bases, Ig2 and Ig3, respectively, from which they are derived. Azine derivative D3 dose-dependently decreased the total cellular glutathione level, suggesting that azine derivatives may exert cytotoxicity by thiol alkylation. Azine derivatives with equal or less cytotoxic potency compared to the mono-Mannich bases they are derived from seemed to be less suitable derivatives for the development of new cytotoxic compounds. PMID- 12736504 TI - Inhibitory effects of caffeic acid phenethyl ester analogues on experimental lung metastasis of murine colon 26-L5 carcinoma cells. AB - We have previously examined the antiproliferative activity of caffeic acid phenethyl ester (CAPE) and its 20 analogues against six tumor cell lines, and found that CAPE analogues possess selective antiproliferative activity toward the murine colon 26-L5 carcinoma cell line. To extend our study, the effects of CAPE analogues on the metastatic development of murine colon 26-L5 carcinoma cells in the lung were examined. The oral administration of CAPE (5 mg/mice/d) for 7 d after tumor inoculation decreased the tumor weight and the number of tumor nodules in the lung by 50% and 50%, respectively, compared to the control, while CAPE (5 mg/mice/d) administered for 7 d before tumor inoculation showed no significant effect. Besides CAPE, 4-phenylbutyl caffeate, 8-phenyl-7-octenyl caffeate, 2-cyclohexylethyl caffeate and n-octyl caffeate at an oral dose of 2 mg/mice/d caused a 55%, 43%, 55% and 35% reduction of the tumor nodules in their lung metastasis formation, respectively. These results further elaborate the possibility of CAPE and its analogues to become a new class of chemopreventive agents for the treatment of colon cancer metastasis. PMID- 12736505 TI - Oxidosqualene cyclases from cell suspension cultures of Betula platyphylla var. japonica: molecular evolution of oxidosqualene cyclases in higher plants. AB - Betula platyphylla var. japonica is a rich source of triterpenoid as it contains dammarane type triterpenes in the leaves, and lupane type and oleanane type triterpenes in the bark. Four oxidosqualene cyclase cDNAs (BPX, BPX2, BPW and BPY) were cloned by homology based PCR methods from cell suspension cultures of B. platyphylla var. japonica. Open reading frames consisting of 2274, 2304, 2268 and 2340 bp were ligated into yeast expression plasmid pYES2 under the control of GAL1 promoter and introduced into lanosterol synthase deficient (erg7) Saccharomyces cerevisiae strain GIL77. Analyses of in vitro enzyme activities and/or accumulated products in the transformants demonstrated that they encode cycloartenol synthase (BPX and BPX2), lupeol synthase (BPW) and beta-amyrin synthase (BPY) proteins. Phylogenetic tree was constructed for all the known oxidosqualene cyclases (OSCs) including the clones obtained in this study, revealing that OSCs having the same enzyme function form respective branches in the tree even though they derive from different plant species. Intriguing correlation was found between reaction mechanism and molecular evolution of OSCs in higher plants. PMID- 12736506 TI - Protective effects of polygodial on gastric mucosal lesions induced by necrotizing agents in rats and the possible mechanisms of action. AB - The effects of polygodial isolated from the leaves of Tasmannia lanceolata on necrotizing agents-induced gastric lesions in rats were compared with capsaicin. Polygodial markedly inhibited the gastric mucosal lesions induced by several necrotizing agents, such as ethanol (ED(50)=0.029 mg/kg, p.o.), 0.6 M HCl (ED(50)=0.26 mg/kg, p.o.), and aspirin (ED(50)=0.38 mg/kg, p.o.), and partly inhibited the gastric mucosal lesions induced by indomethacin, but showed no significant effect on acid output in pylorus-ligated rats at doses of 0.05-0.5 mg/kg. The gastroprotection of polygodial was attenuated by pretreatment with indomethacin (10 mg/kg, s.c.), N(G)-nitro-L-arginine methyl ester (70 mg/kg, i.p.), N-ethylmaleimide (10 mg/kg, s.c.) and ruthenium red (3.5 mg/kg, s.c.). Polygodial (0.2 mg/kg, p.o.) increased the amount of reduced glutathione in gastric mucosa of ethanol-treated group. These results suggested that endogenous prostaglandins, nitric oxide, sulfhydryl compounds and vanilloid receptor mediated effects are involved in the protective effect of polygodial. PMID- 12736507 TI - Simultaneous determination of glycyrrhizin metabolites formed by the incubation of glycyrrhizin with rat feces by semi-micro high-performance liquid chromatography. AB - A method for semi-micro high-performance liquid chromatography (HPLC) has been established for the simultaneous determination of 3alpha-hydroxyglycyrrhetic acid and 3-dehydroglycyrrhetic acid together with glycyrrhizin, glycyrrhetic acid and glycyrrhetic acid mono-glucuronide formed by incubation of glycyrrhizin with rat feces. The analysis was accomplished within 25 min with a TSKgel ODS-80TsQA (150 x 2.0 mm i.d.) column by linear gradient elution using a mobile phase containing aqueous phosphoric acid and acetonitrile at a flow rate of 0.2 ml.min(-1), a thermostatic oven at 25 degrees C, and detection at 254 nm. The detection limits of these compounds were 0.2 pmol per injection (5 microl). The metabolites of glycyrrhizin, by anaerobic or aerobic incubation with rat fecal suspension over 48 h, were determined. Glycyrrhizin was almost completely converted to metabolite glycyrrhetic acid, and metabolites 3alpha-hydroxyglycyrrhetic acid and 3 dehydroglycyrrhetic acid in negligible amounts in anaerobic conditions. However, the metabolic time courses of 3-dehydroglycyrrhetic acid when incubated in aerobic conditions revealed that it apparently continued increasing during the whole incubation period. PMID- 12736508 TI - Bioadhesive delivery of metformin using prosopis gum with antidiabetic potential. AB - The antidiabetic properties of prosopis gum alone and as a bioadhesive base for the delivery of metformin are presented. The bioadhesive value of the gum was commensurate with those of Carbopol 974-P and sodium carboxymethyl cellulose (NaCMC). The release of the drug was higher from prosopis gum based bioadhesive formulations than from NaCMC and Carbopol 974-P products. This was shown by the shorter time required to reach t(50) (the time required for 50% of the drug to be released) or t(20) (time required for 20% of the drug to be released) for the release of metformin. The gum showed moderate antidiabetic properties when used alone. In combination with metformin in a bioadhesive form, the glucose lowering effect was found to be synergistic. The areas under the plasma drug concentration vs. time curves (AUCs) for the bioadhesive combinations were similar to those of the drugs alone in an aqueous system. This shows that the gum did not interfere with absorption of the incorporated drug. However, the areas under the effect vs. time curves (AUECs) were much higher when combined in a bioadhesive form than with the drug alone. The AUCs obtained with NaCMC based bioadhesive formulations were relatively smaller than those of metformin in an aqueous system and the combinations of metformin and prosopis gum. PMID- 12736510 TI - Efficacy of disinfectants and hot water against biofilm cells of Burkholderia cepacia. AB - The effects of various disinfectants and hot water on planktonic cells and biofilm cells of Burkholderia cepacia were investigated. The survival rate of viable B. cepacia cells in suspension decreased to 0.001% or lower within 15 s of exposure to 0.5% benzalkonium chloride, within 30 s of exposure to 0.5% alkyldiaminoethyl glycine, or within 1 min of exposure to 0.1% alkyldiaminoethyl glycine, and decreased to about 0.1% with 60 min of exposure to 0.1% benzalkonium chloride or 0.5% chlorhexidine gluconate, but did not decrease to 1% or less with 60 min of exposure to 0.1% chlorhexidine gluconate. There were no effects of 0.1% and 0.2% chlorhexidine gluconate and 0.1% benzalkonium chloride against biofilm cells of B. cepacia, and 0.5% chlorhexidine gluconate, 0.5% benzalkonium chloride and 0.1% alkyldiaminoethyl glycine were barely effective against biofilm cells even after 60-min exposure. On the other hand, both planktonic cells and biofilm cells of B. cepacia were eradicated within 15 s by sodium hypochlorite, povidone iodine, 80% v/v ethanol, and hot water at 65 degrees C or higher. PMID- 12736509 TI - Microbial viability in preparations packaged for single use. AB - We evaluated microbial viability in preparations packaged for single use only which mandate that residual solution be discarded such as albumin and globulin preparations as blood products, preparations containing albumin (such as urokinase and interferon), fat emulsions, and a preparation containing fat emulsions (propofol). In most preparations, Serratia marcescens and Burkholderia cepacia proliferated rapidly at 30 degrees C. However, in globulin preparations containing 1-2.25% glycine to prevent protein degradation (Gamma-Venin P, Venilon I, Globulin Injection, and Ahlbulin), no growth of S. marcescens and B. cepacia was detected over 24 h at 30 degrees C. For globulin preparations containing 1 2.25% glycine, the injunction to "Discard residual solution after the package has been used" in the package inserts can be revised to "It is possible to use residual solution within 24 h after the package has been used with storage in a cool place." PMID- 12736511 TI - Effects of aspirin and/or salicylate on hydrolysis and glucuronidation of indomethacin in rat erythrocytes and hepatocytes. AB - This study was conducted to explore the mechanism of the pharmacokinetic interaction between aspirin (ASP) and indomethacin (IND) using rat erythrocytes (RBCs) and hepatocytes. ASP was hydrolyzed to salicylic acid (SA) in both the RBCs and hepatocytes. Within RBCs, aspirin and/or salicylate (ASP/SA) increased the concentration of IND, accompanied by a constant hydrolysis of IND. In hepatocytes, a low dose of IND was subjected to glucuronidation rather than hydrolysis, and ASP/SA inhibited both the acylglucuronidation of IND and hydrolysis of IND glucuronide. A high dose of IND underwent hydrolysis with about double the glucuronidation, and ASP/SA decreased the ratio of hydrolysis to glucuronidation, accompanied by a loss of ASP, IND and their metabolites from the medium. Collectively, the results provide metabolic insight into the mechanism of drug-drug interaction between ASP/SA and IND in the hepatocytes and RBCs. PMID- 12736512 TI - Relation between intracellular accumulation and cytotoxic activity of cis-[((1R, 2R)-1, 2-cyclohexanediamine-N, N')bis(myristato)]platinum(II) suspended in Lipiodol. AB - SM-11355, cis-[((1R, 2R)-1, 2-cyclohexanediamine-N, N')bis(myristato)]platinum(II), suspended in Lipiodol (SM-11355/Lipiodol) showed cytotoxic activity in hepatic tumor models in vivo and tumor cell lines in vitro. SM-11355/Lipiodol demonstrated selective retention in tumor tissue in vivo and high accumulation in tumor cells in vitro. This study was aimed to clarify the relation between the cytotoxicity of SM-11355/Lipiodol and intracellular platinum content. The cytotoxic activities were estimated by using WST-1 reagent. Intracellular platinum content and platinum-DNA adduct were estimated following exposure with SM-11355/Lipiodol when methionine was added. Methionine clearly inhibited the cytotoxic activities of SM-11355/Lipiodol. Moreover, intracellular platinum content and platinum-DNA adduct following exposure of SM-11355/Lipiodol decreased with increases in methionine concentration. The characteristic release of SM-11355/Lipiodol was not affected by addition of methionine. The present results suggested that one of platinum compounds exposed to cells following SM 11355/Lipiodol treatment is very similar in cytotoxic mechanism to cisplatin. PMID- 12736514 TI - Antifatigue and antistress effect of the hot-water fraction from mycelia of Cordyceps sinensis. AB - This study was conducted to investigate the chemical component of the hot water (HW) fraction of mycelia of Cordyceps sinensis and its antifatigue and antistress effect against a stimulus in vivo using rats and mice. The growth of mycelia reached a maximum level of 31.6 g/l after 120 h of incubation. The main chemical composition of the HW fraction of mycelia of C. sinensis was found to be carbohydrate (78.9%) with 5% moisture. The swimming endurance capacity of mice orally administered with the HW fraction (150 and 300 mg/kg/d, respectively) was significantly prolonged from 75 to 90 min with a lessening of fatigue. When the HW fraction (150 mg/kg/d) was given to rats for 8 d including a 48 h stress period, the weight changes of the adrenal gland, spleen, thymus, and thyroid, which is an index of stress, were suppressed. The HW fraction also significantly inhibited the increase in total cholesterol and the decrease in alkaline phosphatase levels as biochemical parameters of immobilization stress in rats. PMID- 12736513 TI - Functions of a chitosan-orotic acid salt in the gastrointestinal tract. AB - A chitosan (CS)-orotic acid salt (CS-OT) was prepared, and the release of orotic acid (OT) from CS-OT as well as the adsorption of bile acids by CS-OT was investigated in vitro. The amount of OT released from CS-OT was about 2-2.7 micromol/mg CS-OT and this changed depending on the species of CS. CS-OT also adsorbed bile acids and the amount increased incrementally according to the number of amino group contained in CS. Furthermore, CS-OT was given to rats as feed in order to investigate the influence on serum cholesterol levels. A decrease in serum cholesterol levels was observed in the group, which was fed a diet containing CS-OT or CS for 1-2 weeks, but no differences in body weight changes were recognized. Therefore, CS-OT may be applied to treating hyperlipidemia. PMID- 12736515 TI - Studies on the interactions between drug and estrogen. II. On the inhibitory effect of 29 drugs reported to induce gynecomastia on the oxidation of estradiol at C-2 or C-17. AB - A study was investigated on the inhibitory effect of 29 drugs that have been reported to induce gynecomastia on the 2-hydroxylation of estradiol (E2) by recombinant P450 CYP3A4 and on the 17-oxidation of E2 by hepatic microsomal type II 17beta-hydroxysteroid dehydrogenase (17beta-HSD) of human male. The IC(50) values were determined for each drug relative to the 2-hydroxylation of E2 (catalytic activity: 1.54 nmol/nmol P450/min), and the inhibition constants (K(i)) were determined for 13 drugs of which IC(50) values were 100 microM or less. Ketoconazole exhibited the lowest inhibitory concentration, and IC(50) and K(i) values of 0.007 and 0.01 microM, respectively, were obtained. The IC(50) and K(i) values for each of the 12 remaining drugs were as follows: cyclosporin A (IC(50): 0.064, K(i): 0.30), nicardipine hydrochloride (0.55, 0.29), tacrolimus (0.64, 0.88), mandipine hydrochloride (3.9, 2.6), nisoldipine (10, 3.3), verapamil hydrochloride (10, 20), domperidone (13, 7.2), haloperidol (14, 55), nitrendipine (14, 2.5), chlormadinone acetate (16, 10), flutamide (30, 39) and omeprazole (49, 47). With the exception of cyclosporin A that exhibited a competitive inhibition, the inhibition mechanisms of these drugs were all non competitive. Next, the percentage inhibition of the above 29 drugs relative to the 17-oxidation of E2 (catalytic activity: 0.47 nmol/mg protein/min) was investigated at the approximate therapeutic concentration (1 microM) and at the non-clinical overdose concentration (100 microM). Although none of the drugs investigated exhibited inhibitory effects at a concentration of 1 microM, spironolactone and ketoconazole at 100 microM demonstrated percentage inhibitions of 96% and 77%, respectively. When the K(i) values were determined for these two drugs, the former had a K(i) value of 2.4 microM and the latter, 41 microM, and both of their inhibition mechanisms were non-competitive. On the basis of the above results, a total of 14 drugs consisting of the above 13 drugs plus spironolactone were found to inhibit the 2-hydroxylation or 17-oxidation of E2 in the liver, and this is presumed to act as a trigger that causes as increase in the estradiol pool, followed by induction of gynecomastia. PMID- 12736516 TI - Dynamic responses to acute heat stress between 34 degrees C and 38.5 degrees C, and characteristics of heat stress response in mice. AB - We focused on dynamic responses to acute heat stress between 34 degrees C and 38.5 degrees C. Physiological and neuroendocrinological changes between 34 degrees C and 38.5 degrees C were studied in mice. The influence of humid conditions, 85% relative humidity (RH), on these changes was also investigated. Rectal temperatures increased above 34 degrees C and hematocrit levels increased at 38.5 degrees C 85% RH for 60 min. Food consumption and body weight gains decreased after a daily 60 min exposure to 34, 37 and 38.5 degrees C for 2 weeks. The corticosterone and vasopressin levels in the blood, and catecholamine and serotonin metabolite levels in the hypothalamus were not changed at 34 degrees C, but increased when above 37 degrees C for 60 min. Above 37 degrees C, these physiological and neuroendocrinological changes were accelerated by humid conditions. These results indicated that food consumption and body weight gains decreased above 34 degrees C, and the neuroendocrinological changes, which were accelerated by humid conditions, were induced above 37 degrees C. In comparison with restraint and water immersion stress, heat stress at 37 degrees C 85% RH showed a slower increase in serum corticosterone levels, smaller changes in plasma dopamine and dihydroxyphenylacetic acid levels, and, after repeated exposure, larger decreases in food consumption and body weight gains. This study clarified the relationships between temperature and humidity conditions and physiological and neuroendocrinological changes, along with the characteristics of responses in acute heat stress. PMID- 12736517 TI - Studies on neurosteroids XVI. Levels of pregnenolone sulfate in rat brains determined by enzyme-linked immunosorbent assay not requiring solvolysis. AB - Pregnenolone sulfate (PREGS) is reported to be present in higher concentration in the brain (more than 5 ng/g tissue in the rat) than in blood and is considered to be a neurosteroid. However, there are some doubts on its brain levels, because they were determined by indirect methods (e.g., GC-MS or radioimmunoassay after solvolysis). In the present study, PREGS in rat brains was determined by an enzyme-linked immunosorbent assay, which did not require solvolysis, after pretreatment with an Oasis HLB cartridge. The absolute recovery rate of PREGS through the pretreatment was 60.8%, and the quantitation limit was 33 pg/g tissue for a 200-mg of brain aliquot. Intra- and inter-assay coefficients of variation were less than 15.1 and 9.2%, respectively. The brain PREGS levels in the control rats (n=10) were less than 0.15 ng/g tissue except for one sample (0.42 ng/g tissue) and were lower than the serum levels (n=5, 0.25-0.47 ng/ml). On the contrary, the brain PREGS levels were sufficiently increased after intrapertioneal injection of 2 mg/kg body of PREGS (n=5, 0.37-1.29 ng/g tissue). These results demonstrate that, in rats, the brain PREGS may be derived from peripheral sources, and its actual levels are much lower than those previously measured by indirect methods. PMID- 12736518 TI - Protective effects of quinaprilat and trandolaprilat, active metabolites of quinapril and trandolapril, on hemolysis induced by lysophosphatidylcholine in human erythrocytes. AB - We examined the effects of the angiotensin converting enzyme (ACE) inhibitors captopril, enalaprilat, quinapril, and trandolapril, and their active metabolites quinaprilat and trandolaprilat, on hemolysis induced by lysophosphatidylcholine (LPC) in human erythrocytes. LPC induced hemolysis at the concentrations above the critical micelle concentration (4 microM). Propranolol, used as a reference drug, attenuated the 50% hemolysis induced by 6 microM LPC at concentrations ranging from 100 nM to 100 microM. Similarly, quinaprilat (10 microM) and trandolaprilat (10, 100 microM) significantly attenuated the LPC-induced hemolysis, but other ACE inhibitors did not. Since propranolol possesses a membrane stabilizing action correlated with high lipophilicity, it appears that the high lipophilicity of quinaprilat or trandolaprilat is responsible for the protection from the damage induced by LPC. However, quinapril and trandolapril were not effective, although both drugs have higher lipophilicity than quinaprilat and trandolaprilat. Hence, it is suggested that the high lipophilicity alone may not contribute to the protective effects of ACE inhibitors against LPC-induced hemolysis. None of ACE inhibitors attenuated the hypotonic hemolysis (60 mM NaCl), although propranolol did. Furthermore, neither propranolol (100 microM) nor quinaprilat (50 microM) and trandolaprilat (50 microM) affected LPC micelle formation, suggesting that these drugs do not directly bind to LPC. We therefore believe that the protective effects of quinaprilat and trandolaprilat on the LPC-induced hemolysis may be related physicochemically to their highly lipophilic and ACE inhibitory structures, which probably maintain erythrocyte membrane integrity by a mechanism other than ACE inhibition, prevention of LPC micelle formation or protection against osmotic imbalance. PMID- 12736519 TI - Inhibitory effect of high molecular weight water-soluble chitosan on hypoxia induced inflammatory cytokine production. AB - Chitosan is widely used to treat patients with hypoxia-induced diseases such as ischemia, neuronal death, cerebral stroke, and cerebral infarction. Using the ELISA method, we examined the effect of high molecular weight water-soluble chitosan (WSC) on inflammatory cytokine production in the desferrioxamine (DFX, known to mimic hypoxia)-stimulated human mast cell line HMC-1. DFX significantly increased interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-alpha production compared with the control in a time-dependent manner (p<0.05), but did not affect IL-1alpha production and mRNA expression. The increase in IL-6, IL-8, and TNF-alpha levels was significantly inhibited by WSC in a dose-dependent manner with IC(50) values of 0.77, 0.88, and 2.5 microg/ml, respectively. The maximal inhibition rate of IL-6, IL-8, and TNF-alpha production by WSC was 64+/ 9.7%, 80+/-9.4% and 54+/-4.5%, respectively. In addition, WSC inhibited DFX induced activation of nuclear factor (NF)-kappaB. In conclusion, these results suggest that WSC is an inhibitor of NF-kappaB under hypoxic conditions, which might explain its beneficial effect in the treatment of hypoxia-induced inflammatory diseases. PMID- 12736520 TI - Antipruritic effects of Sophora flavescens on acute and chronic itch-related responses in mice. AB - To find new antipruritic herbal medicines for pruritus, we screened the methanol extracts of seven herbal medicines which have been used to treat dermatologic diseases, testing them on mouse models of acute and chronic itch. When administrated perorally (p.o.) at a dose of 200 mg/kg, methanol extracts of Sophora flavescens and Cnidium monnieri, but not the others, significantly inhibited a serotonin (5-HT)-induced itch-related response (scratching) and the spontaneous scratching of NC mice, a mouse model of atopic dermatitis. The inhibitory effect of Sophora flavescens was stronger than that of Cnidium monnieri. The methanol extract from Sophora flavescens (50-200 mg/kg) inhibited 5 HT-induced scratching in a dose-dependent manner, without any effects on the locomotor activity. These results suggest that Sophora flavescens and its constituents widely affect acute and chronic pruritus, and are possible as new antipruritic agents. PMID- 12736521 TI - Cytoprotective and cytotoxic effects of curcumin: dual action on H2O2-induced oxidative cell damage in NG108-15 cells. AB - The ability of curcumin, a natural antioxidant isolated from Curcuma longa, to inhibit hydrogen peroxide (H(2)O(2))-induced cell damage in NG108-15 cells was examined. When added simultaneously with 500 microM H(2)O(2), curcumin (25-100 microM) effectively protected cells from oxidative damage. However, when the cells were pretreated with curcumin (25-100 microM) for 1.5 h before H(2)O(2) exposure, curcumin was unable to inhibit H(2)O(2)-induced cell damage. Instead, it caused a significant concentration-dependent decrease in cell viability after H(2)O(2) exposure. This dual action of curcumin suggests that pretreatment with curcumin by itself did not have any significant effect on the viability of the NG108-15 cells, but it sensitized them to oxidative damage induced by H(2)O(2) under our experimental conditions. It appears that these events may not relate to the antioxidant and free radical scavenging activities of curcumin. PMID- 12736522 TI - Inhibitory activities of three annonaceous acetogenins on NADH oxidase of chicken liver mitochondria. AB - Annonaceous acetogenins (ACG) are natural products found in the plant family Annonaceae and which strongly inhibited mitochondrial complex I. The inhibition of NADH oxidase of chicken liver mitochondria by three different structural ACG was studied here, and ACG was shown to have potent inhibitory activities similar to rotenone for NADH oxidase. The IC(50) values indicated that bis-adjacent tetrahydrofuran (THF) type squamocin C was more potent than non-adjacent bis-THF type squamostatin B, and the latter was more potent than non-THF type compound 1 in the assay. The roles of structural factors of ACG such as the terminal gamma lactone, the features of other ring moieties and hydroxyl groups, as well as the alkyl chain were simply discussed in this study. PMID- 12736523 TI - Ras transgene expression in TG-AC mice treated with benzo[a]pyrene. AB - Transgene expression and skin tumorigenicity were investigated in transgenic TG AC mice carrying the v-Ha-ras after treatment with benzo[a]pyrene (BP). Animals treated with 40 microg BP (x2/week/mouse) showed 100% tumor response after 25 weeks, as did 40% of the mice treated with 20 microg BP but 10 microg BP did not produce a tumor response. In the case of animals treated with 40 microg BP for 25 weeks, most of the tumors were proven to be carcinomas (80%, 4 out of 5 mice), and all tumors were shown to be positive in terms of transgene expression detected by in situ hybridization. These data suggest that BP was tumorigenic in a dose-dependent manner in TG-AC mice and that TG-AC mice were dependent on transgene expression during BP carcinogenesis. PMID- 12736524 TI - Antioxidant evaluations of novel N-H and N-substituted indole esters. AB - The anti-lipid peroxidation (LP) activity and anti-superoxide formation (SOD) of new N-H and N-substituted indole derivatives were evaluated to determine their antioxidant activity. The results showed that compounds 1, 2, 5, 9 and 12 demonstrate considerable inhibition of lipid peroxidation of mouse liver homogenate. On the other hand, compounds 5 and 12 showed anti-superoxide formation activity at a concentration of 10(-4) M. Consequently, it can be concluded that these compounds exhibit important activity compared with reference compound alpha-tocopherol (vitamin E). PMID- 12736526 TI - The correlation between zeta potential and mucoadhesion strength on pig vesical mucosa. AB - The detachment forces of various polymers are frequently measured to determine their mucoadhesion strength. As the process of mucoadhesion is a consequence of interactions between the mucus layer on mucosa and mucoadhesive polymers, it is greatly dependent on mucus and polymer structure including their charge. It is also known that the glycosaminoglycan layer, which covers the urinary bladder mucosa surface, is highly negatively charged. Therefore, by measuring the zeta potential of polymer dispersions and mucosal homogenates an insight into electrostatic interactions during mucoadhesion can be obtained. In our experiments we chose three polymers, two anionic (polycarbophil, PC; sodium carboxymethyl cellulose, CMCNa) and one cationic (chitosan hydrochloride, CH), for which we expected different zeta potential values and different mucoadhesion strengths. The correlation between the zeta potential and the detachment force was determined. In addition to that, the zeta potential of the scraped surface layer of pig urinary bladders was measured to confirm its negative value. The mucoadhesion strength decreased in the following order: CH>CMCNa=PC. The zeta potentials for all three polymers and for porcine vesical mucosal homogenates were measured in Tyrode solution and two NaCl solutions with different ionic strengths. The lower values of the detachment force correlated well with the more negative zeta potential of the polymer, which might be a consequence of the greater repulsion between negative charges of polymers and glycosaminoglycans. PMID- 12736525 TI - Inhibitory effect of oolong tea on the oxidative state of low density lipoprotein (LDL). AB - In the present study, we investigated the anti-oxidant activity of oolong tea in an oxidation model using human low-density lipoprotein (LDL). Oolong tea suppressed the oxidation of LDL induced by 2-2'-azobis 4-methoxy-2,4 dimethyvaleronitrile (V70) in a dose-dependent manner, that is, it prolonged the lag time to 114.3%, 138% and 199.9% as compared with the control group at 0.5 microg/ml, 1.0 microg/ml, and 2.5 microg/ml, respectively. We also determined the scavenging effect of oolong tea on active oxygen radicals using the electron spin resonance (ESR) technique with 5,5-dimethyl-1-pyrroline N-oxide (DMPO) as a spin trapping agent. The intensity of the ESR signals for the DMPO-OOH adduct formed by the hypoxanthine/xanthine oxidase reaction system with DMPO decreased in the presence of oolong tea. The IC(50) of oolong tea was 19.9 microg/ml. These findings suggested that oolong tea has beneficial effects on health related to its anti-oxidative action. PMID- 12736527 TI - Degradation of flavonoid aglycones by rabbit, rat and human fecal flora. AB - The degradation of thirteen flavonoid aglycones-wogonin, diosmetin, hesperetin, baicalein, morin, genistein, daidzein, quercetin, naringenin, luteolin, kaempferol, apigenin and neophellamuretin-were investigated in rabbit, rat and human fecal flora suspensions as well as in artificial intestinal juice, using high performance liquid chromatography. Separation were performed with a Cosmosil 5C(18)-AR II column by isocratic and gradient elution with 0.1% (v/v) phosphoric acid-acetonitrile as a mobile phase, and detected at 254 nm. The flow rate was 1.0 ml/min. 5,7-Dimethoxycoumarin was used as the internal standard. The result indicated that all flavonoid aglycones except baicalein, diosmetin and quercetin were quite stable in artificial intestinal juice, whereas all were degraded in rabbit, rat and human feces suspension. In rabbit feces, wogonin, diosmetin and hesperetin were less degraded, whereas neophellamuretin, apigenin, kaempferol, luteolin, and naringenin were the most extensively degraded. In rat feces, wogonin and diosmetin were least degraded, whereas kaempferol, quercetin, genistein, luteolin, naringenin and neophellamuretin were extensively degraded. As in human feces, wogonin, daidzein and diosmetin were less degraded, whereas morin, genistein, baicalein, and quercetin were extensively degraded. In conclusion, wogonin and diosmetin were among the less degraded ones for all three feces tested. The presence of a methoxy group on the A or B ring of the flavonoid seems to protect the structure from bacterial degradation. PMID- 12736528 TI - Applications of the Saccharomyces cerevisiae Flp-FRT system in bacterial genetics. AB - The Flp-FRT site-specific recombination system from Saccharomyces cerevisiae is a powerful and efficient tool for high-throughput genetic analysis of bacteria in the postgenomic era. This review highlights the features of the Flp-FRT system, describes current bacterial genetic methods incorporating this technology and, finally, suggests potential future uses of this system. In combination with improved allele replacement methods, recyclable FRT mutagenesis cassettes, whose antibiotic resistance markers can be excised from the chromosome in vivo, are useful for the rapid construction of multiple, unmarked mutations in the same chromosome, and thus aid in the generation of live vaccine strains or food-safe bacteria. The high-specificity of the Flp-FRT system makes it also applicable for manipulation of whole genomes, including in vivo cloning of large genomic segments. Integration-proficient vectors, from which antibiotic resistance markers and replication functions can be evicted after integration of the desired sequences into the chromosome, are useful for the construction of strains destined for environmental release, e.g. strains used as biosensors or for bioremediation. Although the Flp-FRT system is extremely efficient and easy to use, its true potential in bacterial genetics has not yet been fully exploited. On the contrary, in many instances this technology is probably greatly underutilized, especially in gram-positive bacteria. PMID- 12736529 TI - Maize Activator (AC) transposase (TPase) is expressed in Saccharomyces cerevisiae from a genomic clone. detection via Elisa, and proposed use in complementation studies. AB - The maize Activator (Ac) transposase (TPase) was expressed as a Histidine (His) tagged protein in Saccharomyces cerevisiae from a full length genomic clone. Expression was demonstrated via the highly specific nickel-coated Elisa plate method, using an anti-His antibody and 2 separate anti-Ac TPase antibodies, to Ac residues 103-465 and 189-807. AC TPase expression in Saccharomyces is important for two reasons: (a) because the expression from a genomic clone herein permits the future study of RNA splicing mechanisms in common between maize and yeast systems, and (b) because a yeast system can easily be used for demonstrating complementation of function. Thus, such transformed yeast systems could be used in future, to experimentally test whether Ac TPase could complement various yeast mutations. Specifically, Ac TPase may be able to complement (i.e. provide the same function) to yeast transcription factor mutants or to genes mutated in other essential yeast functions. If confirmed, this would lend support to Barbara McClintock's hypothesis that transposable elements can serve as 'controlling elements' within the genome, by their ability to supplement other essential genes' functions, as needed. Work herein is contrasted with existing studies on Ac in yeast. PMID- 12736530 TI - Nucleotide sequence analysis of the sialyltransferase genes of meningococcal serogroups B, C, Y and W135. AB - A rapid method for serogrouping meningococci is essential for the characterization of phenotypically non-groupable meningococcal isolates and clinical samples, particularly for public health management purposes. The Scottish Meningococcus and Pneumococcus Reference Laboratory (SMPRL) provides serogrouping results of meningococcal isolates and clinical samples using a PCR assay which detects restriction fragment length polymorphisms in meningococcal serogroups B, C, Y and W135. Although this PCR system was invaluable when first introduced, it has several drawbacks and lacks the required sensitivity for detecting DNA in clinical samples. Due to the recent introduction of the meningococcal group C conjugate vaccine and an impending group B vaccine, a more robust and informative method for serogroup determination is required. A protocol was devised allowing PCR amplification of the siaD gene of serogroup B, C, Y and W135 meningococci. This system was multiplexed and allowed serogroup differentiation between serogroups B and C and also between B/C and Y/W135 by product size analysis. A nested stage was incorporated into the system for enhanced detection of meningococci in clinical samples, and finally a sequencing protocol was designed allowing detection of any nucleotide changes within the siaD gene. This system allows rapid serogrouping results for use within an agarose gel system as well as more informative results when used for sequencing within the siaD gene. PMID- 12736531 TI - Regulation of FRUA expression during vegetative growth and development of Myxococcus xanthus. AB - Expression of the fruA gene, encoding a putative transcription factor essential for fruiting body formation of Myxococcus xanthus, is specifically activated during development. In the present study, we have analyzed the mechanism of the transcriptional regulation of fruA expression. From gel retardation and footprinting assays using various fruA regulatory regions as probes and competitors, a protein designated factor X was found to specifically bind to a sequence (xbs) located downstream of the transcription-initiation site (+78 to +94) of the fruA gene. Factor X activity was present during vegetative growth and decreased during early development. Analysis of promoter activities of various segments of the fruA regulatory region using the lacZ reporter gene in vivo indicated that a DNA segment extending 45-bp upstream from the transcription initiation site was required for developmentally regulated fruA expression at a low level. In addition, cis-acting regulatory regions located upstream and downstream of the fruA promoter region and including C-box and xbs, were found to be involved in regulation of fruA expression during development. When inserted into the vegA gene, the xbs element inhibited vegA expression during vegetative growth. Together with previously reported results, our studies reveal that fruA expression is regulated by both positive and negative mechanisms during the M. xanthus life cycle. PMID- 12736532 TI - The p1 protein of the yeast transposon Ty1 can be used for the construction of bi functional virus-like particles. AB - Virus-like particles (VLPs) containing heterologous proteins are often used as vaccines. Two approaches for the construction of bi-functional VLPs using hybrid protein pl-380 of the TY1 transposon of Saccharomyces yeast are described. We have shown that both C- and N-termini of p1-380 can be used for the expression of heterologous peptides. Peptides from A. Fumigatus Asp f 2, expressed at the C- and/or N-termini of p1-380, did not interfere with VLP self-assembling, were accessible for antibodies and hence were exposed at the VLP surface. Another way to obtain bivalent VLPs is the formation of mixed particles, which co-express two hybrid pl proteins with different heterologous protein fragments at the C terminus. To do it the yeast cells were transfected with a mixture of two recombinant DNA coding Asp f 2 peptide and green fluorescent protein (Gfp). We have shown that both Asp f 2 peptide and Gfp are expressed within the same particle. To evaluate biological activity of bi-functional VLP a construction containing peptides representing dominant T- and B-cell epitopes of Asp f 2 was produced. Bi-functional particles were more potent in stimulating memory immune responses. These results demonstrate new possibilities of pl-380 based expression system to produce multifunctional VLPs. PMID- 12736533 TI - Comparison of the changes in global gene expression of Escherichia coli induced by four bactericidal agents. AB - DNA microarrays provide a global view of the physiological state of the cell by parallel analysis of the expression levels of all the genes in an organism. The effects of four bactericidal agents on the expression pattern of Escherichia coli MG1655 were assessed. Compounds were chosen on the basis of their different mechanisms of action and included inhibitors of DNA replication and recombination, translation, transcription and cell wall biosynthesis. The addition of rifampin resulted in increased expression of the target, rpoB, as well as several genes involved in nucleotide salvage and purine biosynthesis. The addition of ampicillin resulted in overall changes in gene expression that showed some similarity to changes induced by rifampin. The addition of the antibiotics kanamycin or norfloxacin resulted in the induction of unique gene expression signatures: a heat shock response to kanamycin and an SOS response to norfloxacin. Several genes of unknown function showed expression profiles similar to the genes associated with the SOS or the heat shock response. Thus, these profiles define families of genes with similar expression phenotypes that can be tested for related function. PMID- 12736534 TI - Family of shuttle vectors for ruminal Bacteroides. AB - A family of shuttle plasmids was constructed for genetic transformation of Escherichia coli and of ruminal Bacteroides strains AR20 and AR29. Plasmids were based on the replicon from Bacteroides plasmid pBI191 and were designed for studies of chromosomal integration (pBA), for the identification and study of Bacteroides gene promoters (pPPR) and for the expression of heterologous genes in Bacteroides (pBAC). Electroporation efficiency of Bacteroides was up to 10(5) transformants/microg plasmid, depending on the source of the DNA. The largest plasmid, pBA, was maintained at approximately 8 copies per cell in AR20 and did not measurably alter in vitro growth of transformed cells. In the current work, pBA did not integrate into the chromosomes of AR20 or AR29. The ability of plasmid pPPR to select promoter sequences was demonstrated by removal and replacement of promoters that activate the clindamycin resistance gene. The suitability of pBAC for expression of heterologous genes was demonstrated by expression of the Moraxella species fluoroacetate dehalogenase gene H1 to give intracellular activity of 7 nmol fluoride released/min/mg soluble protein in AR20 and 4 nmol/min/mg in AR29. Spontaneous loss of pBAC under non-selective conditions was 0.11-0.165% per generation, significantly less than loss of the native Bacteroides plasmid pBI191, which was lost at 0.53% per generation. PMID- 12736535 TI - Validation study of a CT-based weighted rating scale for subcortical ischemic vascular disease in patients with mild cognitive deterioration. AB - OBJECTIVES: Subcortical ischemic vascular disease (SIVD) is frequently associated with cognitive impairment. Rating scales to grade cerebrovascular disease are available, but their sensitivity to the clinical features of mild SIVD is unclear. The aim of the study is to devise and validate a computed tomography (CT)-based visual rating scale sensitive to SIVD in patients with mild cognitive deterioration. METHODS: Subjects were 122 consecutive outpatients of a memory clinic (mean age +/- SD 77 +/- 8 years, 71% females, mean +/- SD score of mental state exam 22 +/- 3, 61% with clinical dementia rating of 0.5 and 39% of 1). Diffuse leukoaraiosis, fuzzy and patchy lesions, and lacunes were assessed on ordinary CT films and weights were computed based on clinical indicators of SIVD. A continuous score and an ordinal class (0-3) with higher values indicating higher vascular damage were derived. RESULTS: Intraclass correlation coefficients for intra- and interrater reliability of the subcortical vascular score and class ranged from 0.84 to 0.88. Convergent validity versus the scale of Wahlund et al. was good (Spearman's correlation coefficient between 0.53 and 0.73, p < 0.0005). Known-group validity was assessed versus clinical diagnoses of degenerative (mild cognitive impairment determined by Petersen et al. and NINCDS-ADRDA probable Alzheimer's disease; n = 58), mixed (possible Alzheimer's disease with cerebrovascular disease; n = 21), and vascular cognitive impairment (criteria of Erkinjuntti et al. for subcortical vascular dementia and cognitive impairment; n = 43). Patients with degenerative cognitive impairment were more often in the lower subcortical vascular classes (43% in class 0, 43% in class 1, 14% in class 2, and 0% in class 3), while patients with mixed (0, 24, 24, and 52%) and vascular cognitive impairment were in increasingly higher subcortical vascular classes (0, 12, 35, 53%; p < 0.00005). Criterion-related validity was assessed versus clinical indicators of cerebrovascular disease. Hypertension, platelet aggregation inhibitor use, balance, gait, and bradykinesia increased linearly with increasing subcortical vascular class (p for trend <0.02). CONCLUSIONS: This rating scale is valid and sensitive to capture different degrees of SIVD associated with mild cognitive deterioration. PMID- 12736536 TI - Thymectomy in nonthymoma early-onset myasthenia gravis in correlation with disease severity and muscle autoantibodies. AB - OBJECTIVE: To study the clinical effect of thymectomy in a well-defined early onset MG subgroup and to correlate it to MG severity, the presence of circulating muscle autoantibodies, and the need for pharmacological treatment in a long-term setting. METHODS: Fifty-two consecutive AChR antibody-positive early-onset MG patients (34 thymectomized and 18 nonthymectomized) were included. Severity was assessed and the pharmacological treatment monitored on a yearly basis, starting from the year of MG onset, for 5, 10, 15, and 20 consecutive years; AChR, titin, and RyR antibodies were assayed. RESULTS: In the four follow-up groups, MG severity was significantly higher in nonthymectomized compared to thymectomized MG patients. The postthymectomy MG improvement was significant and persistent. There were 21/34 remissions in thymectomized patients and only 4/18 in the nonthymectomized group. Patients with initially high or low AChR antibody concentration had a similar thymectomy outcome. Only 6 patients had titin antibodies, and none had RyR antibodies. CONCLUSION: The present study indicates a benefit of thymectomy in early-onset MG. The muscle autoantibody concentration does not influence the outcome of thymectomy in early-onset MG. PMID- 12736537 TI - Glutathione S-transferase polymorphisms: susceptibility to migraine without aura. AB - Migraine is considered to be a polygenic multifactorial disease with various environmental and genetic etiologies. We investigated glutathione S-transferase (GST) P1 Ile(105)Val, T1 and M1 polymorphisms in 174 Japanese headache sufferers and 372 Japanese controls. The headache group consisted of 38 cases of migraine with aura, 95 migraine without aura (MWOA) and 41 tension-type headache sufferers. The M1 homozygous deletion genotype was significantly higher in MWOA (64%) compared with controls (46%; p < 0.01; odds ratio = 2.18, 95% confidence interval: 1.32-3.61, adjusted for age and gender). In a comparison of the current smokers, the M1 null frequencies in MWOA were further increased. GSTM1 may be one of the genetic risk factors for MWOA in the Japanese population. PMID- 12736539 TI - Clinical and molecular analysis of chinese patients with thyrotoxic periodic paralysis. AB - Although sporadic thyrotoxic periodic paralysis (TPP) has a much higher prevalence in Asian than in all the other populations studied so far, it is also increasingly being seen at the emergency departments of the West, hence, it is vital to stress the importance of recognizing it. TPP shares some similarities with hypokalemic periodic paralysis (HOKPP). However, the pathophysiology of TPP and the reasons for this higher incidence are not known. We hypothesized that some mutations in the CACNA1S gene, which has been implicated in familial HOKPP, might play a role in TPP. We present 5 Chinese patients who suffer from TPP and demonstrate typical clinical features. No mutation was found on the whole CACNA1S gene. Therefore other molecular mechanisms will have to be examined in order to explain the different TPP incidences. PMID- 12736538 TI - Familial idiopathic brain calcification--a new and familial alpha synucleinopathy? AB - Familial idiopathic brain calcification (FIBC) is a rare disorder characterised by autosomal dominant transmission, adult onset cerebellar and/or extrapyramidal features and idiopathic calcification of the brain. We present a family with FIBC where pathological studies showed that the proband had alpha-synuclein immunopositive glial and neuronal cytoplasmic inclusions in oligodendrocytes in the putamen, midbrain and pons. This may represent a new and familial alpha synuclein disorder causing a predominantly extrapyramidal picture similar to multisystem atrophy. PMID- 12736540 TI - Lactic dehydrogenase isoenzyme in cerebrospinal fluid of children with infantile spasms. AB - Increased levels of lactic dehydrogenase (LDH) in cerebrospinal fluid (CSF) have been reported in association with several intracranial pathologies. We studied LDH isoenzymes in the CSF of children with infantile spasms. CSF samples collected from 12 patients (aged 4-9 months) with infantile spasms were analyzed for total LDH isoenzymes activity, and were compared to samples from 15 normal children. Mean total LDH activity in the CSF was 34.62 +/- 6.52 U/l. Patients with infantile spasms had a lower LDH-1 percentage and higher LDH-3 percentage; the differences from the control group were statistically significant (p < 0.01). LDH-4 and LDH-5 had similar values in both groups. Infantile spasm is apparently associated with a distinct LDH isoenzyme pattern in the CSF. More studies are needed to confirm the rise in LDH-2, LDH-3 and to determine the optimum time of analysis. PMID- 12736541 TI - Transcranial magnetic stimulation study in hereditary spastic paraparesis. AB - The motor-evoked potentials and the cortical excitability by transcranial magnetic stimulation (TMS) were studied in a family with chromosome 2p linked (due to mutations in spastin) and in a family with chromosome 16q linked (due to mutations in paraplegin) hereditary spastic paraparesis (HSP), in order to evaluate the utility of these techniques in identifying the subgroups of the disease. Central motor conduction time and motor treshold to TMS were abnormal in some members of both families; the intracortical inhibition was reduced only in the affected members of the family with chromosome 2p linked HSP, even though the neurological symptoms were sometimes similar and also when clinical features reflecting cortical dysfunction were absent. The motor cortex is differentially involved in the often clinically indistinguishable forms of HSP, and TMS may help in the differential diagnosis. PMID- 12736542 TI - Regional cerebral blood flow and episodic memory in Parkinson's disease: a single photon emission tomography study. AB - In this paper we examined 46 nondemented Parkinson's disease (PD) patients by means of perfusion single photon emission computed tomography (SPECT) and neuropsychological testing. The aim was to detect correlations of regional cerebral blood flow with episodic memory performance, using an operator independent technique for the analysis of SPECT data. A significant positive correlation was found between prefrontal blood flow and episodic memory performances. However, age was the most important determinant of memory scores. Age also correlated significantly negatively with prefrontal perfusion. Our methodology also allowed detection of an inverse correlation of left medial temporal lobe perfusion with the memory score. This had not been found in previous studies and might indicate compensatory mechanisms in the brain of PD patients. It is concluded that episodic memory in nondemented PD patients is most dependent on the effects of aging and that the aging effects on cerebral perfusion in the PD brain parallel to a large extent the findings in normal controls. PMID- 12736543 TI - Bilateral optic ischemic neuropathy related to chronic hepatitis C-associated anticardiolipin antibodies. PMID- 12736544 TI - Treatment options in vertebrobasilar dolichoectasia--case report and review of the literature. PMID- 12736545 TI - High-dose corticosteroid treatment is associated with an increased risk of developing cerebral venous thrombosis. PMID- 12736546 TI - Nonionic contrast neurotoxicity after coronary angiography mimicking subarachnoid hemorrhage. PMID- 12736547 TI - 'Speech initiation hesitation' following subthalamic nucleus stimulation in a patient with Parkinson's disease. PMID- 12736549 TI - [Public health and politics] AB - Not available PMID- 12736550 TI - [Bacterial colangitis: therapeutic features] AB - Cholangitis results from the combination of bactibilia and biliary tract obstruction. In recent years considerable progress has been made in the diagnosis and treatment of cholangitis; advances in endoscopic techniques and antibiotic therapy have ameliorated the prognosis of cholangitis. The choice of an antimicrobial regimen for cholangitis should take into account the antibiotic sensitivities of bacteria colonizing biliary tree, the antibiotic excretion into bile and whether biliary obstruction or bacteremia is present. Successful treatment depends on relieving biliary obstruction and administering antibiotics effective against bacteria implicated. The initial therapy should be active against E. coli and Klebsiella spp., while it is controversial whether the empirical antibiotic regimen should also include coverage against Enterococcus, Pseudomonas and anaerobes. The ureidopenicillins are the preferred initial treatment; the combination piperacillin-tazobactam may be active against the resistant species. Second generation cephalosporins like cefamandole and cefoxitin are still useful, cefoperazone gives excellent coverage against gram negative bacteria, while cefepime may be suitable as treatment for acute cholangitis. In severe cholangitis an aminoglycoside can be added to the beta lactamin; once-daily aminoglycoside administration is associated with a reduced incidence of nephrotoxicity also in patients with cholestasis. Whether the fluoroquinolones are effective in treatment for cholangitis has not been fully evaluated. In patients with suppurative cholangitis prompt endoscopic drainage is mandatory, since antibiotics alone will not sterilize the biliary tract in the face of obstruction. Antibiotic prophylaxis to prevent cholangitis after ERCP should be administered particularly to patients in whom biliary drainage is expected to be difficult; antimicrobial prophylaxis with piperacillin effectively prevents ERCPinduced cholangitis. Antibiotic maintenance therapy can be highly successful in the treatment of recurrent cholangitis in patients with a compromised biliary tract. PMID- 12736548 TI - Recurrence of spontaneous subdural haematoma revealing acquired haemophilia. PMID- 12736551 TI - [New treatment perspectives for community acquired infections] AB - Respiratory and urinary tract infections, of great importance both for their high incidence and for their potential severity (particularly in the lower respiratory tract), represent one of the problems that doctors have to deal with in clinical practice. In outpatients the antibiotic prescription for such pathologies is most often empirical, and needs to consider the local epidemiology, including the incidence of antibiotic resistance, as well as the characteristics of drugs to be prescribed. From this point of view, the lack of drugs with a wide antibacterial spectrum, covering both classical and atypical bacteria, is now overcome by new fluoroquinolones, as levofloxacin. Its characteristics, such as the antibacterial activity extended to Gram positive (particularly S. pneumoniae), the pharmacokinetic profile allowing once daily administration and proper switch therapy, and safety, have been confirmed in wide international clinical studies and make levofloxacin an innovative drug with great potential PMID- 12736553 TI - [Saquinavir + Zidovudine + Lamivudine in HIV positive naive or not naive patients] AB - The aim of the study was to assess the virological/immunological outcome and safety of Saquinavir (SQV), Zidovudine (AZT) and Lamivudine (3TC) therapy in HIV patients. We retrospectively evaluated the charts of 36 HIV patients (male 87% median age 31 yrs, IVDA 72%) who underwent therapy with 5QV (1800 mg/die), AZT (500 mg/die) and 3TC (300 mg/die). 26 (72%) patients were previously on antiviral therapy for at least 16 weeks. No patient ranked in the CDC group. Patients were treated for at least 48 weeks. Hematological indices, changes in CD4+ cell count and plasma HIV-RNA levels were evaluated every 30, 60, and 90 days respectively. At baseline, CD4 cell count and viral load were 230/mcl and 4.55 log respectively for patients previously on therapy and 382/ l and 4.88 log respectively for naive patients. At week 48 the mean CD4+ cell count increase was 70/ l among previously treated and 98/mcl among untreated patients. In respect to the baseline, lower mean HIV-RNA levels were observed at week 12, 24 and 48. No patient showed AIDS defining events. No patient discontinued therapy due to suspected toxicity. Patients showed a good immunological and virological response, regardless of their previous antiviral treatment. The terapy regimen was safe and well tolerated PMID- 12736552 TI - [Prevalence of Toxocara spp in public playgrounds in a town of Central Italy] AB - In this study, the prevalence of Toxocara spp. was evaluated in the soil of 22 public playgrounds of Ancona (Italy). The public playgrounds were selected according to the different areal distribution (centre, low-lying areas centre, hilly areas suburbs) within the urban area of Ancona, central Italy. In six playgrounds there were signs restricting access for dogs; all of them were frequented by children. Parasites were found in the soil samples from 21 out of 22 playgrounds (95.5%). The most prevalent helminth found was Toxocara canis, the common roundworm of dogs. It was detected in the soil samples from 12 (54.5%) playgrounds. This study showed a low frequency of the other parasites. Our data highlight the need for improved knowledge of the relationship between humans and stray animals. Indeed several zooparasites have a role in the etiopathogenesis of common human diseases. The high environmental contamination frequency found underlines the necessity of prompt preventive public health measures, such as control of stray animals, treatment of infected pets and hygiene education of the population to increase awareness of potential zoonotic hazards. PMID- 12736554 TI - [Surveillance of antibiotic-resistant microorganisms for the rational use of antimicrobial drugs] AB - An epidemiological survey of 703 bacterial strains isolated in Pordenone was performed in the first eight months of 1998, to verify the predominant bacterial incidence, the susceptibility to antimicrobial drugs in this period, and variations in the resistance patterns. The study demonstrated some interesting variations in this period: for example, there is a considerable increase in antimicrobial resistance of Escherichia coli (especially to cotrimoxazole, ampicillin, piperacillin and ciprofloxacin) and Pseudomonas aeruginosa (to aztreonam and ciprofloxacin). The results suggest the importance of constant epidemiological surveillance in the hospital and community setting. PMID- 12736555 TI - [Necrotizing fasciitis: case reports and review of the literature] AB - Necrotizing fasciitis is a rare, rapidly progressing infection affecting the superficial fascia and the subcutaneous tissue, accompanied by severe systemic toxicity and multiorgan failure. It is caused by aerobic and anaerobic bacteria, occasionally in a synergistic polymicrobial combination (Type I Necrotizing Fasciitis); in other cases group A -haemolitic Streptoccoccus is the organism responsible for the infection (Type II Necrotizing Fasciitis). The infection often originates from small traumatic injuries or operative wounds and rapidly spreads especially in individuals with identifiable risk factors or immunocompromised patients. Sometimes necrotizing fasciitis occurs when no known portal of entry for bacteria is present. The increasing incidence of necrotizing fasciitis observed may reflect a resurgence of highly virulent mutant strains of group A beta-haemolitic Streptococcus. The pathogenesis, clinical features and treatment of the disease have been reviewed in the light of recent literature. We also report clinical data for four patients with necrotizing fasciitis. They show the importance of early diagnosis and rapid, aggressive and radical surgical intervention. High-dose broad-spectrum antibiotic therapy and intensive medical support are also required to avoid a fatal outcome. PMID- 12736556 TI - [Streptococcus bovis as an opportunistic pathogen during advanced HIV disease] AB - Three rare case reports of HIV-related Streptococcus bovis bacteremia are described, and discussed on the basis of a literature review. The association with an advanced immunodeficiency, leukopenia-neutropenia, and liver cirrhosis is underlined as well as the possibility of multiple recurrences and the lack of apparent gastrointestinal lesions as risk factors. PMID- 12736557 TI - Bacterial esophagitis in patients with HIV disease. AB - The authors carried out a retrospective study by reviewing all patients with HIV disease presenting esophageal symptoms who were evaluated by upper endoscopy. Three cases of bacterial esophagitis are reported and discussed according to literature data. PMID- 12736559 TI - 21st International Congress of Chemotherapy in Birmingham. AB - Not available PMID- 12736558 TI - The use of antidiphterial serotherapy in Bologna in 1895. A pilot experience. AB - It was the year 1894 and the international scientific community was experiencing a historic moment. It was around this time that Pasteur, Kock, L efler, Yersin and Behring discovered the microbiological causes of commonly occurring infectious diseases. In Bologna it was decided to eradicate the diphtheria epidemic which had claimed 152 victims, mainly children, the previous year by experimentally adopting specific serotherapy successfully applied in France shortly before. Supported by local government funding and with the aid of some enterprising local doctors, an efficacious serum was soon produced in Bologna. Good clinical results were obtained and production and sale costs were able to be considerably reduced. Little more than a century later, this is an example of applicative research where there is much still to be revealed. PMID- 12736560 TI - Molecular action of the estrogen receptor and hormone dependency in breast cancer. AB - The measurement of estrogen receptor (ER)alpha in breast cancer tissues is important to discriminate between the hormone dependent and independent tumors. Recently, a second ER, referred to as ERbeta, has been identified. The DNA binding domain of ERbeta is 96% conserved compared with ERalpha, and the ligand binding domain shows 53% conserved residues, suggesting that both receptors can bind estrogen responsive elements on target genes, and that they may also bind similar ligand. While both receptors bind to 17beta-estradiol with equal affinity, other compounds bind with varying affinities to the two receptors. Since the function of ERbeta in breast cancer progression is not well understood, further characterization of the function of ERbeta and its isoforms in breast cancer is warranted. Various kinds of cofactors, such as steroid receptor coactivator-1 (SRC-1), transcription intermediary factor 2 (TIF2), and amplified in breast cancer 1 (AIB1), have also been reported. These coactivators interact with nuclear receptors in a ligand-dependent manner and enhance transcriptional activation by the receptor via histone acetylation/methylation and recruitment of additional coactivator, such as CREB binding protein (CBP)/p300. Thus, action of estrogen is not as simple as thought previously, and is likely influenced by ERbeta, its variants and interaction with cofactors. Improved understanding of the ER mechanism may follow from the discovery of these proteins, although their precise roles remain to be determined. PMID- 12736561 TI - Significance of immunohistochemical assessment of steroid hormone receptor status for breast cancer patients. AB - The assessment of steroid hormone receptors in resected breast cancer tissues is essential to decide whether endocrine therapy is indicated and to select the best treatment for each patient on the basis of receptor status. Both enzyme immunoassay (EIA) and immunohistochemistry (IHC) have been generally used as methods for examination of estrogen receptor (ER) and progesterone receptor (PgR). In some patients, receptor status cannot be examined for various reasons. A questionnaire survey in Japan clarified that ER status is not examined in approximately 40% of patients receiving breast conserving surgery. To eliminate "receptor unknown" cases, IHC examination on paraffin-embedded tissue is useful to assess the in situ receptor status. The concordance rate of ER and PgR status between EIA and IHC is very high and a study of 88 cases revealed a 97.7% concordance for ER and 92.0% for PgR at a cutoff point of 10%. The cutoff point of IHC is controversial and some studies demonstrated that patients showing 1% ER positive cancer cells would benefit from endocrine therapy. On the other hand, immunohistochemical expression of receptors is heterogeneous and some patients with ER negative invasive tumors have ER positive intraductal components. A study of 65 breast cancers demonstrated that ER positive intraductal components were detected in 3.1% cases of ER negative invasive lesions. According to these results and the recommendation of the St. Gallen International Conference, IHC is thought to be more useful than EIA in the assessment of steroid hormone receptor status for breast cancer patients. PMID- 12736562 TI - Current status of endocrine therapy for breast cancer. AB - Endocrine therapy is usually indicated prior to chemotherapy as the first line therapy for metastatic breast cancer patients because of its milder toxicity. Patients who respond to a first-line endocrine therapy have a high chance of responding to a second-line endocrine therapy, and thus the responders to a first line endocrine therapy would better be treated with second or third-line endocrine therapy. In the adjuvant setting, tamoxifen (antiestrogen) has been proven to improve the prognosis of both pre- and postmenopausal estrogen receptor positive breast cancer patients, and goserelin (LH-RH agonist) has been proven to improve prognosis in premenopausal women comparable to chemotherapy (CMF). Very recently, preliminary results have indicated that anastrozole (aromatase inhibitor) is superior to tamoxifen as adjuvant treatment for estrogen receptor positive postmenopausal breast cancer patients. In addition, the recent success of tamoxifen in a chemoprevention trial seems to have ushered in a new era wherein prevention of breast cancer is much more emphasized than treatment of established breast cancer. PMID- 12736563 TI - Endocrine-resistant breast cancer: underlying mechanisms and strategies for overcoming resistance. AB - Estrogen plays important roles in the development and progression of breast cancer. However, one-third of breast cancers fail to respond to endocrine therapy and most endocrine-responsive breast cancers subsequently become resistant to endocrine therapy. A tremendous effort has been made to elucidate the mechanisms responsible for the development of endocrine-resistance in breast cancer. Since the main target molecule of estrogen in breast cancer is estrogen receptor (ER) alpha, most studies have focused on investigating quantitative and qualitative changes in ER-alpha in endocrine-resistant breast cancer. Breast cancers expressing no ER-alpha fail to respond to endocrine therapy. Some breast cancers expressing ER-alpha also fail to respond to endocrine therapy and most breast cancers with acquired endocrine resistance retain ER-alpha expression, which suggests that the disappearance of ER-alpha in breast cancer cells is not a common cause of resistance to endocrine therapy. Recent molecular biological studies have shown evidence that qualitative and functional changes, such as gene mutations and phosphorylation of ER-alpha, cause endocrine resistance in breast cancer. In addition, it has been suggested that endocrine resistance could be induced by epigenetic changes, such as hypoxia, in breast cancer tissues. Understanding the precise mechanisms that underlie endocrine resistance may enable clinicians to develop new strategies for retarding or overcoming endocrine resistance in breast cancer. PMID- 12736564 TI - Higher plasma vascular endothelial growth factor levels correlate with menopause, overexpression of p53, and recurrence of breast cancer. AB - PURPOSE: Vascular endothelial growth factor (VEGF) is an important factor involved in angiogenesis. Many studies have reported that the expression of VEGF in breast cancer is an unfavorable prognostic factor. However, there are few studies that have analyzed blood VEGF levels because most used serum VEGF, generally thought to originate from platelets. We measured plasma VEGF levels, which evaluate the level of tumor-derived VEGF, in various breast diseases. METHOD: We analyzed 15 patients with benign breast disease, 187 patients with primary breast cancer, 32 patients with no postoperative recurrence, and 56 patients with recurrence. Plasma VEGF levels were measured by ELISA. RESULTS: Plasma VEGF levels were higher in malignant than in benign breast disease, and were also high in patients with recurrence or distant metastasis. In primary cases, higher VEGF levels were clearly correlated with menopause and overexpression of p53, and postmenopausal patients with high levels had a significantly lower disease-free survival rate. CONCLUSION: These results suggest that plasma VEGF levels in breast cancer have a clinical significance in that they are associated with the extent or metastasis of malignant lesions and are involved in angiogenesis in postmenopausal patients. PMID- 12736565 TI - Late skin and subcutaneous soft tissue changes after 10-gy boost for breast conserving therapy. AB - PURPOSE: To evaluate the influence of boost irradiation for breast conserving therapy on skin and subcutaneous tissue. MATERIALS AND METHODS: Between 1989 and 1995, 468 patients were treated with breast conserving surgery (quadrantectomy or wide excision with axillary dissection) followed by 50 Gy whole breast irradiation. Among them, fifty-eight patients with positive or close margins were treated with 10 Gy external beam boost irradiation. Skin and subcutaneous soft tissue changes during 5-years of follow-up were examined by inspection and palpation and evaluated using the BCT follow-up form based on EORTC late effect toxicity scoring. RESULT: Four percent (20/468) of the patients showed grade 2 late changes in skin and soft tissue. Four of them had skin telangiectasis, which was limited to within the boost field. Boost irradiation had no definite influence on other late changes. Patients' age, extent of surgery, and pT size had no significant relation to the late changes. The cosmetic score 5 years after BCT was not significantly different between the patients with and without boost irradiation. CONCLUSION: Although 10 Gy boost irradiation after 50 Gy whole breast irradiation increased skin telangiectasis, the late skin and soft tissue changes caused by the boost irradiation were generally mild and there was no substantial deterioration of cosmetic outcome. PMID- 12736566 TI - Ten-year results of a randomized trial on adjuvant chemo-endocrine therapy with tamoxifen for stage II breast cancer. AB - BACKGROUND: A prospective randomized multi-center study was undertaken for 2 years and 3 months from November 1982, with the aim of examining the significance of using a combination of futraful (FT) and tamoxifen (TAM) for postoperative adjuvant therapy for stage II breast cancer after curative surgery. METHODS: Patients were divided into two groups and received one of the following treatment protocols: treatment A, intravenous administration of doxorubicin (DOX) 20 mg/body on the day of surgery and 10 mg/body the next day, followed by oral FT 600 mg/day for 2 years from the 14th day after surgery; treatment B, the same pattern of DOX administration followed by combined therapy with FT and TAM 20 mg/day for 2 years. The number of patients was 428 (treatment A 210 and treatment B 218), of whom 418 (97.7%) were followed for 10 years for analysis. RESULTS: Significantly higher 5- and 10-year overall survival (OS) rates were observed with treatment B compared with treatment A (p=0.0101 and 0.0219). Node-positive patients appeared to derive more benefit from TAM than node-negative patients. The difference in 10-year OS between treatment A and treatment B was more evident than that of the 5-year OS in patients with more than 4 positive nodes (p=0.0313 vs. 0.0479). No increase in adverse reactions was seen as a result of combining TAM with FT. CONCLUSION: The study results demonstrate that for stage II breast cancer concomitant administration of FT and TAM is superior to FT alone for postoperative adjuvant therapy, and administration of TAM for 2 years may contribute not only to 5-year survival rates but also to 10-year survival rates of node-positive patients. PMID- 12736567 TI - Phase I study of docetaxel and cyclophosphamide in patients with advanced or recurrent breast cancer. AB - BACKGROUND: A phase I clinical study of combination chemotherapy with docetaxel and cyclophosphamide (CPA) was performed to determine the maximum tolerated dose (MTD), the incidence and severity of toxicities and the pharmacokinetics in patients with advanced or recurrent breast cancer. METHODS: Docetaxel was administered by intravenous drip infusion over 60 minutes, followed by intravenous bolus injection of CPA every 3-4 weeks. The dosage of docetaxel/CPA was 40/200, 40/400, 50/400, or 60/400 mg/m(2)/day. RESULTS: Fifteen patients were enrolled and received a total of 33 cycles of the combined therapy. The dose limiting toxicities (DLTs) were leukopenia, neutropenia and thrombocytopenia. The MTD was estimated to be docetaxel 60 mg/m(2) in combination with CPA 400 mg/m(2) per day. Plasma clearance of both drugs was similar regardless of dose. The recommended doses of docetaxel/CPA for a phase Utrial are 50/400 mg/m(2)/day every 3-4 weeks. CONCLUSION: The MTD of this combined therapy was docetaxel 60 mg/m(2) and CPA 400 mg/m(2). Neutropenia and leukopeina were common and severe. It is important to stress the need for modification of the dosing scheme. PMID- 12736569 TI - Anastrozole-induced remission of diaphragmatic paralysis from metastatic breast cancer treated with multiple prior therapies. AB - We describe a woman with metastatic breast cancer treated with multiple rounds of prior cytotoxic and endocrine therapies, who presented with ipsilateral diaphragmatic paralysis. The use of anastrozole, a highly selective nonsteroidal aromatase inhibitor (1 mg daily), successfully induced remission of metastatic tumors in our patient, who is partially recovering from diaphragmatic paralysis. PMID- 12736568 TI - Histological classification of invasive ductal carcinoma and the biological parameters in breast cancer. AB - BACKGROUND: The histological classification of invasive ductal carcinoma proposed by the Japanese Breast Cancer Society is based on the appearance of breast cancer invasion. However, few studies have been done to clarify the relationship between the histological classification and other parameters that represent the biological characteristics of individual breast cancer cells. MATERIALS AND METHODS: We analyzed 1,025 invasive ductal carcinomas, consisting of 424 papillotubular, 330 solid-tubular and 271 scirrhous cases, with regard to the status of estrogen receptor (ER), progesterone receptor (PgR), DNA ploidy, epidermal growth factor receptor (EGFR) and p53 protein. RESULTS: Absence of ER and PgR, aneuploidy, EGFR positivity and p53 protein positivity were all observed significantly more frequently in solid-tubular tumors than in the other two types. In addition, the number of abnormal features with regard to these 5 biological parameters was also significantly higher in solid-tubular tumors than in the other two types. CONCLUSION: Abnormalities in the biological parameters listed above were frequently found in the solid-tubular type of invasive ductal carcinoma. PMID- 12736570 TI - Mucocele-like tumor of the breast caused by ductal carcinoma in situ: a case report. AB - A mucocele-like tumor (MLT) of the breast caused by clinging and micropapillary carcinoma in situ is reported. The tumor was a smooth-contoured mass located near the nipple in the lower inner quadrant of the left breast. Ultrasonography showed a cystic tumor and distended ducts. An aspiration specimen contained abundant mucinous material and a small amount of epithelial cell components, but was insufficient for cytological diagnosis. An excisional biopsy specimen disclosed multiple cysts containing mucin with extravasation into the stroma. Additional pathologic diagnosis using consecutive microscopic sections demonstrated ductal carcinoma in situ. Subcutaneous mastectomy was subsequently performed. Histopathologic findings in the resected and biopsy specimens indicated a diagnosis of MLT caused by widely spreading clinging and micropapillary ductal carcinoma in situ. The ductal epithelium of the MLT in an open biopsy specimen must be carefully examined using consecutive microscopic sections to detect occult DCIS. PMID- 12736571 TI - Synchronous bilateral noninvasive ductal carcinoma of the male breast: a case report. AB - A 45-year-old man developed a bloody discharge from his right nipple. Physical examination revealed bloody discharge from his left nipple also but no swelling, breast mass, or axillary lymph nodes. He then underwent bilateral total glandectomy without axillary dissection. Histological examination revealed low grade ductal carcinoma in situ (DCIS) with a low-papillary and cribriform pattern measuring about 4 mm in diameter in the breast bilaterally. To our knowledge, this is the first report of synchronous bilateral DCIS in a male. Since this patient's hormonal profile showed a relatively high blood level of prolactin, the causative relationship between hyperprolactinemia and male breast cancer is discussed. Including our case, 5 of 6 cases reported thus far have been bilateral, and 4 of the 6 cases have been synchronous. We emphasize that the contralateral breast should also be tested or followed in male breast cancer patients with hyperprolactinemia. PMID- 12736572 TI - Severe infusion reaction induced by trastuzumab: a case report. AB - We report a case of a severe infusion reaction caused by trastuzumab. A 59-year old woman with metastatic breast cancer was treated with trastuzumab. During the first infusion, initial symptoms such as severe headache and general fatigue developed. Blood pressure fell 90 minutes after these initial symptoms. A collapsed lung was demonstrated by chest X-ray and computed tomography. Steroid therapy was successfully used for these reactions. Careful monitoring of vital signs, examination of the respiratory system, and the use of steroids are recommended for severe infusion reaction. PMID- 12736573 TI - Metastatic breast cancer of HER2 scored 2+ by IHC and HER2 gene amplification assayed by FISH has a good response to single agent therapy with trastuzumab: a case report. AB - We report that single agent therapy with trastuzumab had a significant effect on metastatic breast cancer, which was confirmed to be HER2 positive by Herceptest showing 2+staining, and gene amplification positively detected by FISH analysis. A 48-year-old woman underwent extended radical mastectomy (T2N0M0 stage II). Three years after the operation supraclavicular lymph node metastasis was noted. Bone scintigraphy showed metastases to the left ribs 5 years after operation. She was treated with chemo-endocrine therapy, but nonetheless could not bear the back pain caused by the bone metastases. Another chemotherapy course could not be permitted because of leukopenia. Immunohistochemistry (IHC) analysis with Herceptest showed 2+staining for HER2 and FISH analysis showed gene amplification of HER2. We started single agent therapy with trastuzumab and she subsequently had remarkably improved back pain. Physical examination and ultrasonography showed disappearance of the previous palpable supraclaviclar lymph nodes. Serum tumor markers were also reduced after the first administration of trastuzumab. The patient is currently alive, with no further progression of the lymph node or bone metastases. PMID- 12736574 TI - Tuberculosis of axillary lymph nodes with primary breast cancer. AB - A rare case of tuberculosis of axillary lymph nodes occurring with primary breast cancer is presented. A 78-year-old woman with no history of pulmonary tuberculosis was admitted to our hospital to undergo examination for a lump in her right breast. The tumor was in the upper outer quadrant of the right breast. On palpation, the tumor was 1.2 cm in diameter and axillary lymph node swelling was noted. Mammography disclosed a spiculated mass and swelling and calcification of the axillary lymph nodes. Sonography showed an irregular hypoechoic mass in the right breast and lymph node swelling in the right axilla, indicating breast cancer with axillary lymph nodes metastases. Chest X-ray showed clustered calcifications in the right axilla and a granular shadow in the right upper lobe. Breast conserving therapy was carried out. Invasive papillotubular carcinoma of the right breast and granulomas with calcification of lymph nodes, compatible with tuberculosis, was diagnosed. Tubercle bacillis were detected by culture of lymph nodes. This case suggests that X-ray is useful for diagnosing lymph node tuberculosis. Lymph node tuberculosis should be suspected when lymph node swelling is noted and X-ray shows clustered calcifications in axillary lymph nodes. PMID- 12736575 TI - [Place of the surgery in the management of brainstem cavernomas. Results of a multicentric study]. AB - OBJECTIVE: To compare the outcome between operated and non-operated brainstem cavernomas. METHOD: Clinical and radiological data of 30 patients harboring 35 brainstem cavernomas were retrospectively reviewed. The mean follow-up period was 47 months. Relationship between the cavernoma and the pi maer was graded. The histology of each operative case was systematically confirmed. The Karnofsky mean score was calculated at admission and at the end of the follow-up period. Patients were classified in three groups according to their outcome grade (Group I=good; Group II=unchanged; Group III=worse). We compared the outcome between operated and non-operated patients. For operated cases, we found several factors of poor outcome. Statistical tests used to compare the groups of patients were Yates modified chi(2) with calculation of the exact probability. Chosen risk was 5%. RESULTS: Thirty-eight percent of operated patients were permanently disabled after surgery whereas the same proportion was improved. None of the patients in the control group worsened at the mean 47-month follow-up despite the occurrence of two hemorrhagic events. Statistical study showed that treatment modality affected clinical outcome in these patients. Only patients presenting with multiple deficits and progressive neurological deterioration were improved with surgery. Surgical access through the floor of the fourth ventricle was correlated with a poor outcome. Partial removal of the cavernoma increased the risk of future hemorrhage. CONCLUSION: Surgery showed no proof of its efficiency in the management of brainstem cavernomas at the end of the follow-up period. Indications of surgical treatment must be restricted to cases with a severe and progressive neurological deterioration. PMID- 12736576 TI - [Epidermoid cyst of the cerebellopontine angle. A surgical series of 10 cases and review of the literature]. AB - Epidermoid cyst or cholesteastoma is a congenital slow growing lesion. It usually arises in the paramedian cisterns of the posterior fossa. Its incidence varies between 0.2 and 1% of all intracranial tumors. The cerebello-pontine angle (CPA) is the most common localization. Our study reports 10 observations of CPA epidermoid cysts, treated in our department between 1989 and 1999. The age of our patients ranged from 20 to 45 years, with male predominance. The patients were admitted with symptoms of cerebello-pontine angle syndrome or signs of posterior fossa tumor. CT-scan was performed in all cases and MRI was performed in 5 patients. The audiometric explorations only provided an orienting contribution. Treatment was exclusively surgical. The retro-sigmoid approach was used in 6 cases, the sub-occipital in 30% and a subtemporal approach in one case. Total resection of the epidermoid cysts was accomplished in 4 cases, and subtotal resection in 4 cases. Two patients underwent partial resection and underwent a revision procedure. The histological examination showed an epidermoid cyst in all cases. The clinical course was controlled in 6 patients: 4 patients recovered and became symptom free. A persisting neurologic deficit was observed in two cases (hearing loss, dysphonia); 4 patients were lost to follow-up. Epidermoid cyst is a benign tumor. Total resection is the ideal treatment, but we have to be aware, taking into consideration the adherence of the tumor to neurovascular structures, of the risks at attempting total resection. PMID- 12736577 TI - [Traumatic spondylolisthesis of the axis: arguments in favor of surgical management after analysis of 8 patients]. AB - BACKGROUND AND PURPOSE: The purpose of this report is to assess the efficacy of primary surgical stabilization in the management of traumatic C2 spondylolisthesis. MATERIAL AND METHODS: Eight patients including 5 men and 3 women (mean age, 44 years) were treated surgically for traumatic spondylolisthesis of the axis. Three patients presented permanent neurological deficit. The indication for operative treatment included fracture instability and association with either neurological deficit or multiple trauma. External immobilization was attempted and failed in two patients. In all cases the procedure was performed by the anterior route and consisted of surgical fixation with C2/C3 discectomy, intersomatic graft placement, and bone fusion. RESULTS: Using the Effendi and Levine classification, fractures were classified as type I in 2 cases, type II in 3, type IIa in 2, and type III in 1. The two patients with severe tetraparesis presented spinal cord contusion at the C2/C3 disk level. Postoperative radiography demonstrated C2/C3 fusion in all patients. No surgical complications were observed. The average hospital stay was 4.5 days. CONCLUSION: Surgical stabilization by C2-C3 fusion via the anterior route is effective for management of all types of traumatic C2 spondylolisthesis. It achieves immediate stability. We recommend its use as a primary intervention not only for unstable lesions but also for lesions associated with neurological deficits or multiple trauma. A major benefit is to facilitate nursing care and patient rehabilitation. PMID- 12736578 TI - [Melanotic schwannoma, a tumor with a unpredictable prognosis: case report and review of the literature]. AB - A large sporadic melanotic schwannoma was found at the C2-C3 level in a 66-year old woman who presented a pyramidal syndrome of all four limbs. The diagnosis was established by histology and immunohistochemical studies and confirmed by ultrastructural examination. Thirty-six cases of melanotic schwannoma located in the spinal cord have been reported in the literature, allowing an analysis of the different histological, clinical, and therapeutic aspects. Usually considered as benign tumor, melanotic schwannoma can recur and progress to malignancy, requiring long-term follow-up, specially in patients with multiple tumors and/or Carney's syndrome. PMID- 12736579 TI - [Primitive intracranial adenoid cystic carcinoma. A case report]. AB - We report the case of a 50-year-old woman presenting a primitive adenoid cystic carcinoma of the middle part of skull base diagnosed by transphenoidal biopsy. She was treated by exclusive radiation therapy with cobalt 60 1.25 MeV and the dose received was 66 Gy in 33 fractions. Our patient remains alive 50 months after the completion of treatment. Adenoid cystic carcinoma is a slow growing malignant tumor with perineural invasion which arises more frequently from salivary glands. The occurrence of primary intracranial adenoid cystic carcinoma is exceptional. The best treatment is radical surgery followed by radiotherapy. Chemotherapy is rarely used. Prognosis is worse than for salivary glands. PMID- 12736580 TI - [Acute subdural hematoma of the posterior fossa in the child. Case report]. AB - Acute subdural hematoma (ASDH) of the posterior fossa is very rare in childhood. It represent 0.52% of all subdural hematomas. We report a case of post-traumatic acute subdural hematoma of the posterior fossa in a 4-year-old child who was drowsy, without a neurologic defect. The CT scan revealed a simple ASDH, with a good recovery after surgical treatment. The literature is reviewed, clinical and therapeutic aspects are discussed. PMID- 12736581 TI - [Bilateral chronic subdural hematoma: spontaneous intracranial hypotension?]. AB - We report a case of bilateral chronic subdural hematoma (SDH) in a 48-year-old man, who presented with postural headaches, tinnitus and progressive confusion without intoxication, head trauma or abnormal hemostasis. Magnetic resonance imaging revealed cerebellar tonsillar herniation in the foramen magnum and a deformation of the brainstem. Outcome was normal after surgery. We discuss about the rare causes of SDH in young adults. PMID- 12736582 TI - [Image-guided epidural blood patch as effective treatment of intracranial hypotension. A case report]. AB - We report the case of a patient with postural headache. A CT scan revealed bilateral subdural hygroma. Brain MRI showed diffuse pachymeningeal enhancement. A diagnosis of intracranial hypotension was therefore made. Thoracic cerebrospinal fluid leak was proved by radionuclide cisternography and contrast myelography. Conservative medical treatment was ineffective. Two thoracic epidural blood patches with radiographic control were made. We think the blood patch is the most important element for success. PMID- 12736583 TI - [A new national post-graduate diploma for neurosurgeons: DEA in Surgical Sciences]. PMID- 12736584 TI - [As always, human quality takes precedence over equipment quality. Reflections on quality assurance of screening mammography and the ineluctable evolution towards digital systems]. PMID- 12736585 TI - [Reduction of acute intestinal intussusception: when and how?]. AB - After sonographic diagnosis, non-surgical therapy for intussusception in children uses pneumatic or hydrostatic (barium or water soluble) reduction. The purpose of this paper, in the light of our experience, is to answer the questions raised by general radiologists in emergency activity about the technique, indications, sedation and risks. PMID- 12736586 TI - [Local recurrence and metastatic dissemination of renal cell carcinoma: clinical and imaging characteristics]. AB - The goal of this pictorial essay is to review the typical and less typical appearances of recurrent and metastatic renal cell carcinoma. Spiral computed tomography is the method of choice to evaluate the postsurgical nephrectomy site and usual sites of metastases. PMID- 12736588 TI - [Follow up of total hip prothesis: storage phosphor versus screen-film radiography]. AB - PURPOSE: Digital radiography may lead to interpretation difficulties in patients with hip prosthesis, especially when initial evaluation was performed using screen film radiography. The purpose of this study was to evaluate inter-observer and inter-technique reproducibility comparing digital and screen-film radiography. MATERIAL AND METHODS: Twenty-eight patients with unilateral (n=24) or bilateral (n=4) hip prosthesis (total=32 prostheses) underwent screen-film and digital (storage phosphor) radiography. The bone-cement and metal-cement interfaces were evaluated for the presence of a lucent line in 14 different sectors reclassified into 3 regions. This evaluation was performed by two independent readers. Inter-observer and inter-technique reproducibility were calculated using kappa values. RESULTS: The inter-observer reproducibility was negligible, poor or average with Kappa values ranging from 0.07 to 0.47. The degree of agreement between the two methods was poor, average or good for both observers with Kappa values ranging from 0.10 to 0.66. CONCLUSION: Although inter technique agreement is not high, it is better than inter-observer agreement. Therefore, storage phosphor technique should not be rejected for total hip prosthesis follow-up, even though previous films were screen-film radiographs. PMID- 12736587 TI - [Functional anatomy of the central nervous system]. AB - The central nervous system (CNS) has a particular regional functional anatomy. The morphological support of cognitive functions can now be depicted using functional imaging. Lesions of the central nervous system may be responsible of specific symptoms based on their location. Current neuroimaging techniques are able to show and locate precisely macroscopic lesions. Therefore, the knowledge of functional anatomy of the central nervous system is useful to link clinical disorders to symptomatic lesions. Using radio-clinical cases, we present the functional neuro-anatomy related to common cognitive impairments. PMID- 12736589 TI - [Low-dose chest CT with millimetric thin slices: myth or reality?]. AB - PURPOSE: To evaluate the diagnostic accuracy of millimetric thin slices low dose chest CT. MATERIALS AND METHODS: Forty one patients underwent a chest CT thin slices (1 mm every 10 mm) exploration using both a 170 milliamperage and a low dose acquisition using 80 mA. The examination were read by 2 senior radiologists specialized in chest imaging without knowledge of acquisition parameters and in a random order. A statistical analysis of interobserver agreement was performed using Kappa analysis. Doses of both acquisition were estimated by compagning the dose length product calculated by the CT software and be using a simulation software. RESULTS: Excellent interobserver and intermodalities agreements were found. A 53% decrease in dose was estimated with the low dose modality compare to the normal dose. CONCLUSION: Low dose thin slice chest CT using 80 mA has a similar diagnosis accuracy as standard dose thin slice chest CT and delivers half dose of irradiation. PMID- 12736590 TI - [Swallowing study with kinetic MRI using a single shot fast spin echo sequence in healthy volunteers and patients treated for head and neck cancer]. AB - PURPOSE: To evaluate the single shot fast spin echo sequence (SSFSE) rapid MRI sequence for swallowing study, to describe the swallowing phases analyzable by kinetic MR Imaging and to show the advantages and the limits of the method in patients treated for head and neck cancer. METHODS AND MATERIALS: A preliminary study was conducted in 8 healthy volunteers and 11 patients treated for a head and neck cancer between June 1999 and April 2001. Examinations were obtained on a 1.5 T machine using a multi-slice SSFSE sequence with an acquisition time of 1 second/slice. The different phases of swallowing were analyzed. Examinations were done with and without water ingestion. RESULTS: Kinetic MRI allows to clearly analyze the anatomy and the dynamic of the tongue, the soft palate, the hyoid bone, the larynx and the posterior pharyngeal wall. The hypopharyngeal and esophageal phases were suboptimally assessed. CONCLUSION: Kinetic MRI allows to clearly analyze the oral and early pharyngeal phases of swallowing, which can be correlated with normal and surgically reconstructed areas in spite of the low temporal resolution and the non-physiologic position. It is simple and rapid to perform and may be useful in treated patients with head and neck cancer. PMID- 12736591 TI - [Radiodermatitis in interventional radiology (hand dose measurement, screening and compensation)]. AB - It is to be feared that the expansion of interventional radiology could lead to the reappearance of chronic radiodermatitis. Only a few studies mention the doses received by the hands, however these radiation doses are always high. Whereas clinical examinations lead to a belated diagnosis, capillary microscopy, a quick and non invasive test, enables to detect vascular injuries at the subclinical stage. Radiological protection is currently based on leaded gloves and on the reduction of exposure duration. Compensation for chronic radiodermatitis depends on the practitioner's status. In spite of a better evaluation of the overall irradiation, hand exposure remains a cause for concern. Capillary microscopy can provide appropriate medical surveillance. PMID- 12736592 TI - [Splenic rupture following colonoscopy: report of an unusual case in the absence of underlying splenic disease]. AB - Splenic injury following colonoscopy is rare and, to our knowledge, 18 cases only have been reported so far in the literature. The presumed mechanisms for splenic injury during colonoscopy are direct trauma to the spleen or excessive traction of the splenocolic ligament this latter mechanism being due to decreased mobility between the spleen and the colon secondary to adhesions. Splenomegaly or underlying splenic disease are two conditions which are considered to be predisposing factors to splenic injury. We report one case of splenic rupture which was due to colonoscopy in the absence of any underlying splenic disease. This case was diagnosed and monitored by computed tomography, thus avoiding unnecessary splenectomy. PMID- 12736593 TI - [A peculiar form of neonatal adrenal insufficiency: the IMAGe association. Two new cases]. AB - We report two siblings with an IMAGe syndrome. IMAGe is a newly reported syndrome characterized by the association of intra-uterine growth retardation, metaphyseal dysplasia, congenital adrenal hypoplasia and genital anomalies. This clinical association has only been described in five unrelated males. These two additional patients (one brother and one sister) suggest an autosomal recessive inheritance although identification of new cases will give further insight into the pathogenesis. The radiologic signs of osteopenia and metaphyseal dysplasia can suggest this diagnosis in affected individuals. PMID- 12736595 TI - [Pericardial hydatid disease: three case reports]. AB - Hydatid disease of the heart is very rare, representing about 0.2 to 2% of all cases. Most hydatid cysts of the heart are located within the left ventricular wall. Pericardial location is also very rare and patients present with various symptoms. US and CT have a primary role in the diagnostic workup of this disease. PMID- 12736594 TI - [Enterolith ileus: a rare complication of small bowel diverticulosis]. AB - An elderly patient with subacute small bowel obstruction due to an enterolith that evolved within a small bowel diverticulum is reported. Presence of small bowel diverticulum is not rare. But small bowel obstruction secondary to an enterolith formed within a small bowel diverticulum is a rare complication. Enterolith ileus closely resembles gallstone ileus in its clinical presentation. Diagnosis can be established only by documenting the absence of aerobilia and the presence of small bowel abnormality causing stasis, like small bowel diverticulosis. PMID- 12736596 TI - [Quid? Pancreatic metastasis of a left renal neoplasm removed in 1996]. PMID- 12736597 TI - Pediarix--a combined vaccine for infants. PMID- 12736598 TI - Oxybutynin transdermal (Oxytrol) for overactive bladder. PMID- 12736599 TI - Eplerenone (Inspra). PMID- 12736601 TI - The need to address new threats. PMID- 12736602 TI - Wanna be a hero? Maybe it's time for a reappraisal... PMID- 12736603 TI - GW gets READI. PMID- 12736604 TI - The power of teamwork. PMID- 12736606 TI - MCI challenge. Crash on the 710 Freeway: 41 patients transported in 1 hour 41 minutes at one of the nation's largest multi-casualty traffic accidents. PMID- 12736607 TI - Building MCI capability: how one rural, volunteer, BLS service upgraded its system to triage & care for up to 100 victims during an MCI. PMID- 12736608 TI - Automated MCI patient tracking: managing mass casualty chaos via the Internet. PMID- 12736609 TI - MCI Resource Guide 2003. PMID- 12736610 TI - Femur fractures: complications & treatments of traumatic femoral shaft fractures. AB - Femoral shaft fractures can present many challenges to health-care providers from the time of injury to the time of discharge and beyond. Ongoing assessment and definitive interventions prove successful in limiting additional morbidity and mortality. PMID- 12736613 TI - eJEMS: electronic & data management issues: e-mail basics. PMID- 12736611 TI - Amazing terrorism tool: new foam could revolutionize decon. PMID- 12736614 TI - Aging gracefully. PMID- 12736617 TI - Don't overlook older volunteers. PMID- 12736616 TI - ALS roulette. PMID- 12736618 TI - Complications in infants undergoing surgery for congenital cataract in the first 12 weeks of life: is early surgery better? AB - PURPOSE: To determine if there is an association between the timing of surgical intervention for congenital cataract within the first 12 weeks of life and the prevalence of postoperative complications. METHODS: We performed a retrospective review of records from 1990 to 2000 of infants who underwent surgery for congenital cataract within the first 12 weeks of life. Eighty eyes in 55 children were involved with a minimum follow up of 6 months. Bilateral cataracts were present in 25 and monocular cataracts in 30 infants. A limbal approach lensectomy vitrectomy was performed in all infants. Children with aphakia were rehabilitated with contact lens or glasses. Operative and postoperative complications-including glaucoma, nystagmus, strabismus, retinal detachment, and posterior capsule opacification/secondary membranes-were recorded. Ocular and systemic associations were noted. Statistical analysis was carried out with classification and regression trees (CART). RESULTS: The mean age at the time of surgery was 31.5 +/ 23.3 days (median, 26.5; range, 2 to 84). Mean follow up from the time of surgery was 2.85 +/-1.9 years (median, 2; range, 0.5 to 8). Persistent fetal vasculature (persistent hyperplastic primary vitreous) was present in 14 eyes. One infant with bilateral persistent fetal vasculature had bilateral retinal dysplasia and was excluded from the analysis. Glaucoma developed in 12 infants (22%); nystagmus was present in 18 infants (33%); strabismus developed in 28 infants (52%); and secondary membranes developed in 7 eyes (13%). CART analysis suggests that glaucoma is more prevalent in infants when the surgery was performed between 13.5 and 43 days of life (CART = 0.370); nystagmus when surgery is performed between 48 and 84 days of life (CART = 0.500); strabismus when surgery is performed between 55.5 and 84 days of life (CART = 0.600); and secondary membranes when surgery is performed between 26.5 and 40 days of life (CART = 0.4). CONCLUSIONS: Our data suggest that the first 2 weeks of life comprise the most favorable time for decreasing postoperative complications resulting from surgical intervention for infants presenting with cataracts within the first 12 weeks of life. PMID- 12736619 TI - The association between myopic shift and visual acuity outcome in pediatric aphakia. AB - PURPOSE: The advent of intraocular lens implantation after pediatric cataract surgery necessitates an increased understanding of refractive development. The significant variation in rate and amount of refractive change among eyes, both aphakic and pseudophakic, is well recognized, although the causes of such variation remain unclear. The purpose of this study was to determine if a correlation exists between the rate of refractive growth (RRG) and visual acuity outcome in pediatric aphakia. METHODS: Multicenter, retrospective observational case series. One hundred and twenty-five eyes of 85 patients with cataract surgery before 1 year of age and a minimum of 3 years of follow-up were analyzed. RRG was calculated for each eye using the logarithmic model of ocular growth and compared with final logMAR acuity using linear regression. RESULTS: The correlation of RRG with final logMAR acuity was statistically significant (r(2) = 0.10; P <.01), ie, 10% of variance in RRG is related to acuity outcome. The correlation was higher in unilaterally aphakic patients (n = 44; r(2) = 0.19; P <.01) than in bilaterally aphakic patients (n = 81; r(2) = 0.08; P <.01). Eyes with visual acuity of 20/60 or better had a significantly lower RRG than those with poorer acuity (4.1 v 5.4 diopters (D); P <.01). CONCLUSIONS: RRG in aphakia is correlated with visual acuity outcome. Eyes with poorer acuity have a greater RRG. PMID- 12736620 TI - Visual outcome of cataract surgery in children with congenital rubella syndrome. AB - PURPOSE: To evaluate visual outcome after cataract surgery in children with congenital rubella syndrome (CRS). METHODS: A retrospective analysis was conducted on 40 eyes of 22 children with CRS who underwent cataract surgery. Thirty-six eyes underwent lensectomy with anterior vitrectomy, and 4 eyes underwent extracapsular cataract extraction with primary posterior capsulectomy. The median age at surgery was 6 months, and median duration of postoperative follow up was 68.5 months. Each follow-up visit consisted of visual acuity estimation, refraction, anterior and posterior segment examination, and intraocular pressure measurements. RESULTS: Visual acuity at final follow up was 6/24 or better in 6 (15.0%) eyes, and 22 (55.0%) eyes had visual acuity less than 3/60. Postoperative complications included transient corneal edema in 18 (45.0%) eyes, glaucoma in 5 (12.5%) eyes, after cataract in 1 (2.5%) eye, and hyphema in 1 (2.5%) eye. Ocular disorders affecting visual outcome included stimulus deprivation amblyopia, glaucoma, optic atrophy, corneal opacity, and after cataract. Associated systemic disorders included neurological problems in 15 (68.2%), hearing loss in 12 (54.6%), cardiovascular problems in 9 (40.9%), and speech abnormalities in 7 (31.8%) children. CONCLUSIONS: The less-than-optimal postoperative visual outcome suggests the need to look at primary prevention of rubella, especially in developing countries. PMID- 12736621 TI - The efficacy of superior oblique split Z-tendon lengthening for superior oblique overaction. AB - PURPOSE: To evaluate the efficacy of superior oblique muscle split Z-tendon lengthening in the treatment of superior oblique muscle overaction with respect to normalization of superior oblique ductions and versions, reduction of A pattern strabismus and correction of vertical deviations. In addition, to determine the incidence of complications of the procedure including overcorrection and the effect of the procedure on horizontal deviation. METHODS: A retrospective review was performed of 19 patients who underwent superior oblique muscle split Z-tendon lengthening for A-pattern strabismus or significant vertical deviation associated with superior oblique overaction between June 1980 and September 1999. Ten males and 9 females were included, ranging in age from 2 to 45 years (mean: 9 years). Patients underwent either a unilateral or bilateral superior oblique tendon lengthening based on the patient's clinical picture. RESULTS: Ten of 11 patients (91%) who underwent unilateral tendon lengthening and 10 of 16 eyes (63%) that underwent bilateral superior oblique tendon had complete normalization of superior oblique action. Seven of 10 patients (70%) who underwent unilateral or bilateral tendon lengthening for a clinically significant A-pattern had the A-pattern completely eliminated or brought to a clinically insignificant level. All 8 patients who underwent a unilateral tendon lengthening for a clinically significant vertical deviation had reduction of the vertical deviation. There was no statistically significant effect of either the unilateral or bilateral procedure on horizontal deviation. CONCLUSIONS: The superior oblique muscle split Z-tendon lengthening procedure is highly effective for normalization of superior oblique muscle function and reduction of A-pattern strabismus and vertical deviations associated with superior oblique muscle overaction. PMID- 12736622 TI - Measurement of ocular torsion after macular translocation: disc fovea angle and Maddox rod. AB - PURPOSE: To compare two methods of measuring ocular torsion (the subjective Maddox rod [MR] test versus the objective disc-fovea angle [DFA] test) after macular translocation surgery. METHODS: Ocular torsion was measured on consecutive patients after macular translocation at Duke University Eye Center between August 2001 and April 2002. Both MR and DFA measurements of torsion were made at the same clinic visit 4 to 8 weeks after the translocation surgery and again within 3 months after extraocular muscle surgery to decrease torsion. MR and DFA measurements were each performed by a separate examiner who was blinded to the results of the other method. RESULTS: Thirty-five patients (35 eyes) were included for evaluation. Twenty-nine of these patients had intorsion measured by both MR and DFA after macular translocation but before extraocular muscle surgery (MR mean of 40.3 + 7.2 degrees v DFA mean of 47.0 + 7.9 degrees [P <.001]). The intrapatient reproducibility of the MR test was high (using four readings per session), with a mean coefficient variation of 4.8%. Twenty-five patients had residual torsion measured by both MR and DFA after extraocular muscle surgery (MR mean of 4.2 + 4.7 degrees v DFA of mean 4.8 + 7.0 degrees). There was good correlation between MR and DFA measurements of torsion (r(2) = 0.9). CONCLUSIONS: DFA measurement correlates well with MR measurement of torsion in patients after full macular translocation. This study verifies the reproducibility of MR to measure large angles of torsion and offers DFA as a simple corroborative test for measuring ocular torsion in patients with poor vision or cooperation. PMID- 12736623 TI - Acuity, ophthalmoscopy, and visually evoked potentials in the prediction of visual outcome in infants with bilateral optic nerve hypoplasia. AB - PURPOSE: To determine whether Teller Acuity Cards, transient visually evoked potentials (VEPs), and optic disc size estimated from ophthalmoscopy were predictive of acuity outcome in infants and young children with bilateral optic nerve hypoplasia (ONH). METHODS: Twenty-eight infants (mean age, 7 months) with bilateral ONH underwent clinical assessment, including ophthalmoscopy. All but a few of these patients underwent neuroimaging studies, analysis of transient VEPs to multiple stimuli, and repeated acuity assessment. Acuity outcome was assessed, on average, 28 months later. RESULTS: Acuity outcome was significantly correlated with the estimated optic disc diameter, initial acuity, and VEP signal-to-noise ratios (r = 0.80, 0.71, 0.69, respectively; all P <.001). Multiple regression analysis showed that the initial acuity and estimated optic disc diameter accounted for 73% of the variation in acuity outcome. VEPs to white-black gratings segregated infants by 6 months of age, whose acuity outcome was better or worse than 5.6 cycles/degree (20/100). CONCLUSIONS: Acuity outcome was predicted in infants with bilateral ONH with a linear equation using initial acuity and estimated optic disc diameter. Additionally, analysis of VEPs may segregate infants with a good visual outcome from those with a poor visual outcome. Longer follow-up will be necessary to determine final Snellen acuity. PMID- 12736624 TI - Adult strabismus surgery under propofol sedation with local versus general anesthesia. AB - PURPOSE: To compare operating room and hospital discharge times between adult strabismus surgeries using intravenous propofol sedation with local anesthesia versus those using general anesthesia. METHODS: Thirty adult patients underwent uncomplicated strabismus procedures performed by one surgeon using propofol sedation and local subtenon's anesthesia. These were retrospectively matched with adult patients undergoing similar procedures by another surgeon using general inhalational anesthesia. Only one muscle had undergone previous surgery, and no adjustable sutures were used. Times from incision closure to leaving the operating room (Out OR time), and to hospital discharge (DC time) are compared. RESULTS: Propofol/local Out OR times ranged 2 to 8 minutes (mean, 4.8). General anesthesia Out OR times ranged from 3 to 28 minutes (mean, 8.8) (means differ at P<.001). At 10 minutes, 100% of propofol patients left the OR, but only 63% of general patients had done so (P<.001). Propofol/local DC times ranged from 30 to 140 minutes (mean, 64.8). General DC times ranged from 68 to 325 minutes (mean 116.5) (means differ at P<.001). At 60 minutes after completion of surgery, 53% of propofol/local patients had left the hospital, whereas none of general patients had left (P<.001). At 2.5 hours after surgery, 100% of propofol/local patients had left the hospital, but 10% of general patients remained, with two staying more than 5 hours. CONCLUSIONS: Many adult strabismus surgeries may be performed more efficiently with intravenous propofol sedation and local subtenon's anesthesia than with general anesthesia. Times from the end of surgery to leaving both the OR and the hospital are decreased compared with those of general anesthesia. Extreme delays are rare with propofol/local, but they occurred with general anesthesia. PMID- 12736625 TI - Combined use of cryotherapy and diode laser photocoagulation for the treatment of threshold retinopathy of prematurity. AB - PURPOSE: Cryotherapy and indirect laser retinal photoablation are both effective in the treatment of retinopathy of prematurity (ROP). We describe the safety, efficiency, and effectiveness of combined cryotherapy and diode laser photocoagulation to treat threshold ROP. METHODS: Records of patients developing threshold ROP from January 1, 1996 through December 31, 1998, were retrospectively reviewed to identify those treated with combined cryotherapy and photocoagulation and followed up for at least 45 days postoperatively. Diode laser was used to ablate posterior avascular retina, and cryotherapy was used for anterior retina. Data reviewed included ocular and systemic complication rates, treatment duration, number of laser burns, most recent fundus examination, visual acuity, and refraction. RESULTS: In 13 patients, 23 eyes received combined treatment. No intraoperative complications occurred. Mean duration of anesthesia and treatment was 35 +/- 8 minutes/eye. A mean of 117 +/- 84 laser burns/eye were applied. In 20 of 23 eyes (87.0%), anatomic outcome was favorable at last examination. In 13 of 16 eyes (81.3%), functional (visual acuity) outcome was favorable (visual acuity better than 20/200) at 1 year. At 6 months or later, 14 of 16 eyes (87.5%) measured were myopic, of which 5 (31.3%) were highly myopic (> 6 diopters). CONCLUSIONS: The effectiveness of treating ROP with combined cryotherapy and diode laser photocoagulation compares with that of either modality alone. By decreasing the number of laser applications, combined therapy may be faster and technically easier for eyes with very posterior ROP. This may decrease the number of complications seen when either excessive cryotherapy or laser retinal photoablation is used. PMID- 12736626 TI - Computer-automated quantification of plus disease in retinopathy of prematurity. AB - BACKGROUND: In some cases of retinopathy of prematurity (ROP), it difficult to determine with certainty whether plus disease is present or absent. We have developed a computer program that captures digital images from a video-indirect ophthalmoscope, identifies and traces the major posterior pole blood vessels, measures the dilation and tortuosity of each vessel, and calculates whether or not an eye has plus disease. Our purpose was to determine the accuracy of the computer program in comparison with two masked examiners. METHODS: A representative sample of posterior pole images from 20 premature infants, 10 normal and 10 representing various degrees of dilation and tortuosity, was extracted from our video database and analyzed by the computer program as well as by two masked examiners experienced in the diagnosis of ROP. The standard photograph from the Cryotherapy for ROP study, representing the minimum degree of dilation and tortuosity required for plus disease, was also digitized, analyzed, and used as a numeric comparison for the automated determination of plus disease. RESULTS: Of the five images determined to have plus disease by both examiners, four were calculated to have plus disease by the computer program (80% sensitivity). Of the 11 images without plus disease, 10 were calculated not to have plus disease by the computer program (91% specificity). CONCLUSIONS: Our computer program has very good sensitivity and specificity compared with masked examiners' determination of the presence or absence of plus disease. Automated analysis of dilation and tortuosity of posterior pole blood vessels has the potential to remove subjectivity from the determination of plus disease. PMID- 12736627 TI - The incidence of visually impaired children with retinopathy of prematurity and their concomitant disabilities. AB - PURPOSE: To determine, in relation to two previous studies, the incidence of visual impairment (VI) caused by retinopathy of prematurity (ROP) in children born in the Netherlands between 1994 and 2000 and their associated nonvisual disabilities. METHODS: From all institutes, schools, and ambulatory services for the partially sighted and blind, data on ROP infants born between 1994 and 2000 were retrieved. Apart from neonatal and ophthalmologic data, other data were gathered concerning the following concomitant disabilities: behavioral abnormalities, epilepsy, hearing deficit, developmental delay, and distinct neurological handicaps. Birth rate data were obtained from the Central Bureau of Statistics for the Netherlands. RESULTS: Data on 51 of 54 infants (94.4%) were studied. There was a statistically insignificant decrease in the incidence of visually impaired children caused by ROP in the period between 1994 and 2000 compared with the period between 1986 and 1994. Neonatal and ophthalmologic data of the infants were comparable for the study period and the previous period. The incidence of behavioral abnormalities in infants with ROP-related sequelae increased significantly in the study period compared with the previous one (46.9% v 21.8%, P <.05. The percentage of infants who received treatment for severe ROP did not change significantly (56.9% v 43.9% in the previous period). CONCLUSIONS: This study shows a statistically insignificant decrease in the number of infants with visual impairment caused by ROP in the Netherlands in the period between 1994 and 2000 as well as a significant increase in the incidence of behavioral abnormalities in these children. Still, 43.1% of the infants did not receive acute-phase ROP treatment. PMID- 12736628 TI - The polaroid suppression test in a pediatric population with ophthalmologic disorders. AB - PURPOSE: The Polaroid suppression test (PST) is a new method for early detection of amblyogenic factors by screening for suppression. The apparatus can elicit suppression with the use of Polaroid filters. The aim of the present study was to examine a population of children with known ophthalmologic disorders using the PST to determine the rate of false-negative results of the PST. METHODS: Six hundred four children, varying in age between 3 and 15 years (mean, 7.9) were examined using the PST. Ophthalmologic disorders ranged from strabismus and amblyopia to refractive disorders. RESULTS: Mean testing time for the PST was 43 seconds. The PST could not be administered to 34 children (5.6%); 443 children (73.3%) had abnormal results; and 127 children (22.2%) showed no suppression. The suppression in constant strabismus was detected in almost all cases. The sensitivity for accommodative forms of strabismus was lower, but amblyopia was never missed in these cases. In children with normal eye alignment, only 2.7% with an interocular acuity difference of more than 0.1 logMAR had no suppression. Of all 119 children with clinical defined amblyopia, only 1 (0.8%) did not have suppression. Overall sensitivity of the PST for strabismus and/or abnormal interocular acuity difference was 96.2% and specificity was 41.1%. CONCLUSIONS: The PST has great potential as a visual screening tool in young children. Only few children with amblyogenic factors were missed. Thus, the test can differentiate those children at risk for amblyopia from normally sighted children. Because specificity is lower, all children showing suppression with the PST in a screening situation should have further examination by the health care worker before being referred to the ophthalmologist. PMID- 12736629 TI - Poor results after recession of both medial rectus muscles in unilateral small angle Duane's syndrome, type I. AB - PURPOSE: To eliminate an abnormal face turn in unilateral Duane's syndrome, type I, the medial rectus muscle of the Duane's eye is commonly recessed. Additional recession of the normal contralateral medial rectus muscle has been advocated in selected cases, although little has been published regarding this technique. We present poor results in a small consecutive series. METHODS: Four consecutive cases of unilateral Duane's syndrome, type I, with small-angle primary position esotropia are retrospectively reviewed with attention to postoperative face turn. In all cases, the medial rectus muscle of the "normal" eye was recessed as was the medial rectus muscle of the Duane's eye. RESULTS: Two subjects showed little to no improvement in face turn; one subject developed an increased turn; and the last subject developed a consecutive exotropia. CONCLUSIONS: In small-angle Duane's syndrome, type I, recession of the normal medial rectus may decrease the positive effects of recessing the Duane's medial rectus muscle with respect to face turn as well as increase the risk of consecutive exotropia. An alternate theory of normal-eye Duane's surgery is proposed. PMID- 12736630 TI - Traumatic optic nerve injury occurring after forceps delivery of a term newborn. PMID- 12736631 TI - Encephalocraniocutaneous lipomatosis: a neurocutaneous syndrome. AB - The neurocutaneous pattern syndromes are a group of disorders characterized by congenital abnormalities involving both the skin and the nervous system for which no identifiable cause has been isolated.(1) Ophthalmologic manifestations of these syndromes are common. These rare syndromes often have overlapping ophthalmic and systemic findings. Examples include encephalocraniocutaneous lipomatosis (ECCL), oculocerebrocutaneous syndrome (OCC), and linear nevus sebaceous syndrome (LNS). Clarifying the diagnostic criteria for these syndromes is a worthy goal because it will help elucidate the phenotypic spectrum of these poorly understood diseases as well as possibly facilitate genetic counseling. In this short report, a case of ECCL is used to illustrate the clinical manifestations of neurocutaneous syndromes. PMID- 12736632 TI - Altered motivation and learning following opiate withdrawal: evidence for prolonged dysregulation of reward processing. AB - Opiate abuse has been associated with cognitive deficits in human addicts. To determine if prior opiate exposure alters the ability to learn, we trained animals in an instrumental learning task for a food reward. During a 2-week period after withdrawal, morphine-abstinent rats were significantly slower at learning an escalating fixed-ratio response for food reward compared to placebo treated animals. When these same animals were trained in a conditioned suppression paradigm (two tone-shock pairings given in the operant box), the morphine-withdrawn animals showed greater retention by taking significantly longer to resume responding for food reward when the tone was presented. In a third experiment, morphine-abstinent rats withdrawn 2 or 5 weeks were tested for their ability to associate a highly palatable food reward with a specific environment using a place-conditioning paradigm. At 2 weeks postwithdrawal, morphine-abstinent rats did not show any significant place preference for a food they readily consumed, while placebo-treated rats readily learned to prefer the food-paired environment. At 5 weeks postwithdrawal, rats developed significantly less preference for food-associated cues, but more preference for morphine associated cues, compared to placebo-treated animals. These data suggest that prior morphine exposure may have prolonged effects on the motivation for natural rewards, which in turn may compromise the ability of former addicts to overcome their addictions. PMID- 12736633 TI - The phospholipase C-IP3 pathway is involved in muscarinic antinociception. AB - The cellular events involved in muscarinic analgesia were investigated in the mouse hot-plate test. Intracerebroventricular (i.c.v.) pretreatment with antisense oligonucleotides (aODNs) against the alpha subunit of G(q) and G(11) proteins prevented the analgesia induced by physostigmine and oxotremorine. Furthermore, administration of the phospholipase C (PLC) inhibitor U-73122, as well as the injection of an aODN complementary to the sequence of PLCbeta(1), antagonized the increase of the pain threshold induced by both cholinomimetic drugs. In mice undergoing treatment with LiCl, which impairs phosphatidylinositol synthesis, or treatment with heparin, an IP(3) receptor antagonist, the antinociception induced by physostigmine and oxotremorine was dose-dependently antagonized. I.c.v. pretreatment with TMB-8, a blocker of Ca(2+) release from intracellular stores, prevented the increase of pain threshold induced by the investigated cholinomimetic drugs. Coadministration of Ca(2+) restored the muscarinic analgesia in LiCl, heparin, and TMB-8-preatreated mice. On the other hand, i.c.v. pretreatment with the selective protein kinase C (PKC) inhibitor calphostin C, resulted in a dose-dependent enhancement of physostigmine- and oxotremorine-induced antinociception. The administration of PKC activators, such as PMA and PDBu, dose dependently prevented the cholinomimetic drug-induced increase of pain threshold. Neither aODNs nor pharmacological treatments employed produced any behavioral impairment of mice as revealed by the rota-rod and hole board tests. These results indicate a role for the PLC-IP(3) pathway in central muscarinic analgesia in mice. Furthermore, activation of PKC by cholinomimetic drugs may represent a pathway of negative modulation of muscarinic antinociception. PMID- 12736637 TI - Hope for best, prepare for worst. PMID- 12736634 TI - Association of a critical CSF tryptophan threshold level with depressive relapse. AB - This work studies association between relapse during acute tryptophan depletion (ATD) and CSF level of tryptophan (TRP) in remitted depressives treated with sertraline or bupropion. Eight medication-responding depressives ingested an ATD amino acid mixture during 48-h continuous CSF sampling before and after treatment. Mood rating scores were compared with nadir levels of TRP in CSF. CSF TRP nadirs averaged 8.7% of am baselines in remitted patients. Mood relapsed whenever the CSF nadir was below 40 nmol/l TRP in remitted patients, and never when above (Fisher's exact test, P=0.029). Relapsing medication responders also showed very low preantidepressant ATD-induced nadirs. ATD-induced relapses were associated with low CSF TRP levels. Individual susceptibility to depletion may be independent of antidepressant treatment, mood state, or treatment status. Resistance to relapse may invoke an undefined, protective CNS mechanism against extremely low CSF levels of TRP during ATD. PMID- 12736638 TI - Berlin's university crisis. PMID- 12736639 TI - France purges space programme in bid to survive budget crisis. PMID- 12736640 TI - US Army joins hunt for SARS drug. PMID- 12736642 TI - Los Alamos contract open to bids. PMID- 12736641 TI - US fails to quantify threat of West Nile virus. PMID- 12736643 TI - Geneticists question fees for use of patented 'junk' DNA. PMID- 12736644 TI - Physicist takes the reins at Santa Fe complexity centre. PMID- 12736645 TI - Dying cells dragged screaming under the microscope. PMID- 12736646 TI - Homeland science chief wants quick fixes. PMID- 12736649 TI - Weapons proliferation: Asia's nuclear family. PMID- 12736647 TI - AIDS researchers seek criteria for vaccines. PMID- 12736650 TI - Spinal injuries: In search of a miracle. PMID- 12736651 TI - SARS testing: First past the post. PMID- 12736652 TI - Lack of trained security staff delays US visas. PMID- 12736653 TI - How technology can reduce our impact on the Earth. PMID- 12736654 TI - Unbalanced view of a dynamic world. PMID- 12736655 TI - Designer scientific literature. PMID- 12736656 TI - Peer review: recognition via year-end statements. PMID- 12736663 TI - Mathematics: Conjuring with conjectures. PMID- 12736662 TI - Planetary Science: Catch a falling star. PMID- 12736664 TI - Molecular physiology: Tuned for longer life. PMID- 12736665 TI - Prion diseases: Cannibals and garbage piles. PMID- 12736667 TI - Animal behaviour: Homing is a breeze for sea turtles. PMID- 12736668 TI - Cell biology: Patches for wounded muscle. PMID- 12736669 TI - Complex fluids: Spread the word about nanofluids. PMID- 12736670 TI - Computational biology: Biosensor design. PMID- 12736672 TI - Palaeontology: A fly in the biogeographic ointment. PMID- 12736673 TI - Electronic paper: Flexible active-matrix electronic ink display. PMID- 12736674 TI - Comparative genomics: Insecticide resistance in mosquito vectors. PMID- 12736675 TI - Nitrogen storage: UV-B radiation and soil microbial communities. PMID- 12736677 TI - The evolutionary origin of complex features. AB - A long-standing challenge to evolutionary theory has been whether it can explain the origin of complex organismal features. We examined this issue using digital organisms--computer programs that self-replicate, mutate, compete and evolve. Populations of digital organisms often evolved the ability to perform complex logic functions requiring the coordinated execution of many genomic instructions. Complex functions evolved by building on simpler functions that had evolved earlier, provided that these were also selectively favoured. However, no particular intermediate stage was essential for evolving complex functions. The first genotypes able to perform complex functions differed from their non performing parents by only one or two mutations, but differed from the ancestor by many mutations that were also crucial to the new functions. In some cases, mutations that were deleterious when they appeared served as stepping-stones in the evolution of complex features. These findings show how complex functions can originate by random mutation and natural selection. PMID- 12736678 TI - The structure of DNA in the nucleosome core. AB - The 1.9-A-resolution crystal structure of the nucleosome core particle containing 147 DNA base pairs reveals the conformation of nucleosomal DNA with unprecedented accuracy. The DNA structure is remarkably different from that in oligonucleotides and non-histone protein-DNA complexes. The DNA base-pair-step geometry has, overall, twice the curvature necessary to accommodate the DNA superhelical path in the nucleosome. DNA segments bent into the minor groove are either kinked or alternately shifted. The unusual DNA conformational parameters induced by the binding of histone protein have implications for sequence-dependent protein recognition and nucleosome positioning and mobility. Comparison of the 147-base pair structure with two 146-base-pair structures reveals alterations in DNA twist that are evidently common in bulk chromatin, and which are of probable importance for chromatin fibre formation and chromatin remodelling. PMID- 12736679 TI - Photographic observations of Neuschwanstein, a second meteorite from the orbit of the Pribram chondrite. AB - Photographic observations of meteoroids passing through the atmosphere provide information about the population of interplanetary bodies in the Earth's vicinity in the size range from 0.1 m to several metres. It is extremely rare that any of these meteoroids survives atmospheric entry to be recovered as a meteorite on the ground. Pribram was the first meteorite (an ordinary chondrite) with a photographically determined orbit; it fell on 7 April 1959 (ref. 1). Here we report the fourth meteorite fall to be captured by camera networks. We determined the atmospheric trajectory and pre-atmospheric orbit of the object from the photographic records. One 1.75-kg meteorite--named Neuschwanstein and classified as an enstatite chondrite--was recovered within the predicted impact area. The bolide's heliocentric orbit is exceptional as it is almost identical to the orbit of Pribram, suggesting that we have discovered a 'stream' of meteoritic objects in an Earth-crossing orbit. The chemical classifications and cosmic-ray exposure ages of the two meteorites are quite different, however, which implies a heterogeneous stream. PMID- 12736681 TI - Spreading of nanofluids on solids. AB - Suspensions of nanometre-sized particles (nanofluids) are used in a variety of technological contexts. For example, their spreading and adhesion behaviour on solid surfaces can yield materials with desirable structural and optical properties. Similarly, the spreading behaviour of nanofluids containing surfactant micelles has implications for soil remediation, oily soil removal, lubrication and enhanced oil recovery. But the well-established concepts of spreading and adhesion of simple liquids do not apply to nanofluids. Theoretical investigations have suggested that a solid-like ordering of suspended spheres will occur in the confined three-phase contact region at the edge of the spreading fluid, becoming more disordered and fluid-like towards the bulk phase. Calculations have also suggested that the pressure arising from such colloidal ordering in the confined region will enhance the spreading behaviour of nanofluids. Here we use video microscopy to demonstrate both the two-dimensional crystal-like ordering of charged nanometre-sized polystyrene spheres in water, and the enhanced spreading dynamics of a micellar fluid, at the three-phase contact region. Our findings suggest a new mechanism for oily soil removal- detergency. PMID- 12736680 TI - Controlled vesicle deformation and lysis by single oscillating bubbles. AB - The ability of collapsing (cavitating) bubbles to focus and concentrate energy, forces and stresses is at the root of phenomena such as cavitation damage, sonochemistry or sonoluminescence. In a biomedical context, ultrasound-driven microbubbles have been used to enhance contrast in ultrasonic images. The observation of bubble-enhanced sonoporation--acoustically induced rupture of membranes--has also opened up intriguing possibilities for the therapeutic application of sonoporation as an alternative to cell-wall permeation techniques such as electroporation and particle guns. However, these pioneering experiments have not been able to pinpoint the mechanism by which the violently collapsing bubble opens pores or larger holes in membranes. Here we present an experiment in which gentle (linear) bubble oscillations are sufficient to achieve rupture of lipid membranes. In this regime, the bubble dynamics and the ensuing sonoporation can be accurately controlled. The use of microbubbles as focusing agents makes acoustics on the micrometre scale (microacoustics) a viable tool, with possible applications in cell manipulation and cell-wall permeation as well as in microfluidic devices. PMID- 12736682 TI - Redistribution of energy available for ocean mixing by long-range propagation of internal waves. AB - Ocean mixing, which affects pollutant dispersal, marine productivity and global climate, largely results from the breaking of internal gravity waves- disturbances propagating along the ocean's internal stratification. A global map of internal-wave dissipation would be useful in improving climate models, but would require knowledge of the sources of internal gravity waves and their propagation. Towards this goal, I present here computations of horizontal internal-wave propagation from 60 historical moorings and relate them to the source terms of internal waves as computed previously. Analysis of the two most energetic frequency ranges--near-inertial frequencies and semidiurnal tidal frequencies--reveals that the fluxes in both frequency bands are of the order of 1 kW x m(-1) (that is, 15-50% of the energy input) and are directed away from their respective source regions. However, the energy flux due to near-inertial waves is stronger in winter, whereas the tidal fluxes are uniform throughout the year. Both varieties of internal waves can thus significantly affect the space time distribution of energy available for global mixing. PMID- 12736683 TI - Experimental evidence that potassium is a substantial radioactive heat source in planetary cores. AB - The hypothesis that (40)K may be a significant radioactive heat source in the Earth's core was proposed on theoretical grounds over three decades ago, but experiments have provided only ambiguous and contradictory evidence for the solubility of potassium in iron-rich alloys. The existence of such radioactive heat in the core would have important implications for our understanding of the thermal evolution of the Earth and global processes such as the generation of the geomagnetic field, the core-mantle boundary heat flux and the time of formation of the inner core. Here we provide experimental evidence to show that the ambiguous results obtained from earlier experiments are probably due to previously unrecognized experimental and analytical difficulties. The high pressure, high-temperature data presented here show conclusively that potassium enters iron sulphide melts in a strongly temperature-dependent fashion and that (40)K can serve as a substantial heat source in the cores of the Earth and Mars. PMID- 12736684 TI - Universal scaling relations in food webs. AB - The structure of ecological communities is usually represented by food webs. In these webs, we describe species by means of vertices connected by links representing the predations. We can therefore study different webs by considering the shape (topology) of these networks. Comparing food webs by searching for regularities is of fundamental importance, because universal patterns would reveal common principles underlying the organization of different ecosystems. However, features observed in small food webs are different from those found in large ones. Furthermore, food webs (except in isolated cases) do not share general features with other types of network (including the Internet, the World Wide Web and biological webs). These features are a small-world character and a scale-free (power-law) distribution of the degree (the number of links per vertex). Here we propose to describe food webs as transportation networks by extending to them the concept of allometric scaling (how branching properties change with network size). We then decompose food webs in spanning trees and loop forming links. We show that, whereas the number of loops varies significantly across real webs, spanning trees are characterized by universal scaling relations. PMID- 12736685 TI - Defective membrane repair in dysferlin-deficient muscular dystrophy. AB - Muscular dystrophy includes a diverse group of inherited muscle diseases characterized by wasting and weakness of skeletal muscle. Mutations in dysferlin are linked to two clinically distinct muscle diseases, limb-girdle muscular dystrophy type 2B and Miyoshi myopathy, but the mechanism that leads to muscle degeneration is unknown. Dysferlin is a homologue of the Caenorhabditis elegans fer-1 gene, which mediates vesicle fusion to the plasma membrane in spermatids. Here we show that dysferlin-null mice maintain a functional dystrophin glycoprotein complex but nevertheless develop a progressive muscular dystrophy. In normal muscle, membrane patches enriched in dysferlin can be detected in response to sarcolemma injuries. In contrast, there are sub-sarcolemmal accumulations of vesicles in dysferlin-null muscle. Membrane repair assays with a two-photon laser-scanning microscope demonstrated that wild-type muscle fibres efficiently reseal their sarcolemma in the presence of Ca2+. Interestingly, dysferlin-deficient muscle fibres are defective in Ca2+-dependent sarcolemma resealing. Membrane repair is therefore an active process in skeletal muscle fibres, and dysferlin has an essential role in this process. Our findings show that disruption of the muscle membrane repair machinery is responsible for dysferlin-deficient muscle degeneration, and highlight the importance of this basic cellular mechanism of membrane resealing in human disease. PMID- 12736686 TI - Pathogenic bacteria attach to human fibronectin through a tandem beta-zipper. AB - Staphylococcus aureus and Streptococcus pyogenes, two important human pathogens, target host fibronectin (Fn) in their adhesion to and invasion of host cells. Fibronectin-binding proteins (FnBPs), anchored in the bacterial cell wall, have multiple Fn-binding repeats in an unfolded region of the protein. The bacterium binding site in the amino-terminal domain (1-5F1) of Fn contains five sequential Fn type 1 (F1) modules. Here we show the structure of a streptococcal (S. dysgalactiae) FnBP peptide (B3) in complex with the module pair 1F12F1. This identifies 1F1- and 2F1-binding motifs in B3 that form additional antiparallel beta-strands on sequential F1 modules-the first example of a tandem beta-zipper. Sequence analyses of larger regions of FnBPs from S. pyogenes and S. aureus reveal a repeating pattern of F1-binding motifs that match the pattern of F1 modules in 1-5F1 of Fn. In the process of Fn-mediated invasion of host cells, therefore, the bacterial proteins seem to exploit the modular structure of Fn by forming extended tandem beta-zippers. This work is a vital step forward in explaining the full mechanism of the integrin-dependent FnBP-mediated invasion of host cells. PMID- 12736687 TI - Nicotinamide and PNC1 govern lifespan extension by calorie restriction in Saccharomyces cerevisiae. AB - Calorie restriction extends lifespan in a broad range of organisms, from yeasts to mammals. Numerous hypotheses have been proposed to explain this phenomenon, including decreased oxidative damage and altered energy metabolism. In Saccharomyces cerevisiae, lifespan extension by calorie restriction requires the NAD+-dependent histone deacetylase, Sir2 (ref. 1). We have recently shown that Sir2 and its closest human homologue SIRT1, a p53 deacetylase, are strongly inhibited by the vitamin B3 precursor nicotinamide. Here we show that increased expression of PNC1 (pyrazinamidase/nicotinamidase 1), which encodes an enzyme that deaminates nicotinamide, is both necessary and sufficient for lifespan extension by calorie restriction and low-intensity stress. We also identify PNC1 as a longevity gene that is responsive to all stimuli that extend lifespan. We provide evidence that nicotinamide depletion is sufficient to activate Sir2 and that this is the mechanism by which PNC1 regulates longevity. We conclude that yeast lifespan extension by calorie restriction is the consequence of an active cellular response to a low-intensity stress and speculate that nicotinamide might regulate critical cellular processes in higher organisms. PMID- 12736688 TI - Computational design of receptor and sensor proteins with novel functions. AB - The formation of complexes between proteins and ligands is fundamental to biological processes at the molecular level. Manipulation of molecular recognition between ligands and proteins is therefore important for basic biological studies and has many biotechnological applications, including the construction of enzymes, biosensors, genetic circuits, signal transduction pathways and chiral separations. The systematic manipulation of binding sites remains a major challenge. Computational design offers enormous generality for engineering protein structure and function. Here we present a structure-based computational method that can drastically redesign protein ligand-binding specificities. This method was used to construct soluble receptors that bind trinitrotoluene, l-lactate or serotonin with high selectivity and affinity. These engineered receptors can function as biosensors for their new ligands; we also incorporated them into synthetic bacterial signal transduction pathways, regulating gene expression in response to extracellular trinitrotoluene or l lactate. The use of various ligands and proteins shows that a high degree of control over biomolecular recognition has been established computationally. The biological and biosensing activities of the designed receptors illustrate potential applications of computational design. PMID- 12736689 TI - Direct observation of catch bonds involving cell-adhesion molecules. AB - Bonds between adhesion molecules are often mechanically stressed. A striking example is the tensile force applied to selectin-ligand bonds, which mediate the tethering and rolling of flowing leukocytes on vascular surfaces. It has been suggested that force could either shorten bond lifetimes, because work done by the force could lower the energy barrier between the bound and free states ('slip'), or prolong bond lifetimes by deforming the molecules such that they lock more tightly ('catch'). Whereas slip bonds have been widely observed, catch bonds have not been demonstrated experimentally. Here, using atomic force microscopy and flow-chamber experiments, we show that increasing force first prolonged and then shortened the lifetimes of P-selectin complexes with P selectin glycoprotein ligand-1, revealing both catch and slip bond behaviour. Transitions between catch and slip bonds might explain why leukocyte rolling on selectins first increases and then decreases as wall shear stress increases. This dual response to force provides a mechanism for regulating cell adhesion under conditions of variable mechanical stress. PMID- 12736695 TI - Cash crisis in the corridor. PMID- 12736690 TI - Folding at the speed limit. AB - Many small proteins seem to fold by a simple process explicable by conventional chemical kinetics and transition-state theory. This assumes an instant equilibrium between reactants and a high-energy activated state. In reality, equilibration occurs on timescales dependent on the molecules involved, below which such analyses break down. The molecular timescale, normally too short to be seen in experiments, can be of a significant length for proteins. To probe it directly, we studied very rapidly folding mutants of the five-helix bundle protein lambda(6-85), whose activated state is significantly populated during folding. A time-dependent rate coefficient below 2 micro s signals the onset of the molecular timescale, and hence the ultimate speed limit for folding. A simple model shows that the molecular timescale represents the natural pre-factor for transition state models of folding. PMID- 12736696 TI - Science has flourished in eastern Germany since reunification, bringing researchers from far and near. But the tide of investment may be turning. PMID- 12736698 TI - [Recanalizing therapies in acute stroke]. AB - Systemic thrombolysis with rt-PA within 3 hours after symptom onset has developed into a standard therapy for acute stroke patients. Recombinant t-PA is now licensed in Europe for this indication. Within 3 to 6 hours, no general therapeutic standard has been established yet. Currently, the available data are in favor of intra-arterial thrombolysis for patients with proximal occlusions of the middle cerebral artery. Systemic thrombolysis is not generally recommended in this time window, but is used in some centers for selected patients. In addition to thrombolysis and new strategies to improve thrombolysis (including patient selection with MRI), several interventional and surgical recanalization techniques are discussed. PMID- 12736699 TI - [Why to operate on the carotid artery?]. AB - AIM, METHODS, PATIENTS: In a prospective interdisciplinary study of 468 carotid artery stenoses we compared the ultrasound plaque morphology with the histological diagnosis. RESULTS: For detection of soft plaques and hard plaques by preoperative ultrasound examination we found a sensitivity of 80%. The accuracy for the estimation of the degree of the stenosis was 98%. In case of stroke in progression and completed stroke we found ulcerations by histological examination in 72%. By ultra-sound these ulcerations were detected only in 53%. We found a significant higher figure for TIA and stroke in soft plaque patients than in asymptomatic stage, in which hard plaque formation was seen more often. CONSEQUENCES: Patients with a soft plaque seem to have a higher risk of a postoperative permanent neurological deficit. However, the dangerous plaque formation with thrombosis or ulceration was not be detected with appropriate accuracy. Therefore the risk of embolisation cannot be predicted sufficiently by means of duplex scanning. This would be extremely important for the indication for stenting in this area. The number of HITS (high intensity transient signals) is 8 to 20 times higher in carotid stenting. PMID- 12736701 TI - [Stents in the iliac arteries: a minimally invasive method]. AB - AIM: We demonstrate patients to reveal the conditions for good results with respect to acute and long term out-come of interventional therapy of iliac artery occlusion. MATERIAL AND METHODS: 225 Patients with aortoiliacal occlusion who underwent surgery or intervention were evaluated. All patients were discussed in an interdisciplinary angiologic conference. 63 patients primary had an operation, 161 patients primary were treated by intervention. The two cohorts were compared concerning their structure, the primary success and the primary and secondary patency over one year. RESULTS: Both cohorts showed significant differences, the surgical treated patient had a higher clinical stage as well as more complex iliacal occlusion. The interventional treated patients had higher comorbidity. Primary technical success and patency rates did not show any difference. The complication rate was equally low in both cohorts. In percutaneous treated patients after predilatation, embolization into the periphery occurred. With primary stent PTA, there was no embolization. CONCLUSION: The percutaneous recanalization also of long iliac occlusion is a save, successful alternative to surgery. Nevertheless, there are limits. For optimal therapy management an interdisciplinary consultation is necessary. To avoid embolization, after recanalization primary stent PTA should be performed. PMID- 12736700 TI - [The unstable carotid stenosis: definition and biological processes]. AB - The occurrence of cerebral or retinal ischemic symptoms ipsilateral to high-grade internal carotid artery (ICA) stenosis indicates a status of instability with a substantial risk for future major stroke. Additionally, the detection of microembolic signals downstream of ICA stenosis is predictive for future cerebral ischemia in asymptomatic and symptomatic patients. There is substantial evidence that in unstable ICA stenosis plaque rupture and thrombus formation are the most frequent pathoanatomic findings. In contrast, in nearly the half of unstable carotid plaques the lumen surface appears to be intact. Within plaque tissue, the unstable plaque is mainly characterized by a substantial amount of inflammatory cell (i. e. macrophages, T-cells) infiltration. These cells are mainly localized in the fibrous cap near the necrotic core. Produced by macrophages, matrix degrading enzymes (e. g. MMP-9) are overexpressed in the unstable ICA stenosis. Thrombogenicity is mainly determined by the local concentration of activated tissue factor, also expressed by inflammatory cells. Furthermore, a significantly higher rate of apoptotic smooth muscle cells can be found within the fibrous cap of instable carotid stenoses. Whether infection with Chlamydia pneumoniae contribute to instability is unlikely, because a positive association to clinical instability has not been shown up to now. The exact and detailed characterization of the unstable ICA plaque and the correlation of different biological mechanisms to clinical instability may offer the possibility to use it as a human model of unstable atherosclerosis in general and to test the efficacy of new developed anti-atherosclerotic pharmaceutical agents. PMID- 12736702 TI - [Stenting of the femoral artery: the angiologist's perspective]. AB - The percutaneous transluminal angioplasty of stenosis and short occlusions of the femoral artery is a well established, safe and effective treatment option. The additional placement of stents during angioplasty is associated with frequent and rapid restenosis due to intimal hyperplasia. Therefore, stent placement of the femoral artery is commonly recommended only for special circumstances, including PTA-induced dissection of the vessel wall, elastic recoil, and eccentric stenosis. Placement of stents near joints, e. g. in the common femoral artery or the popliteal artery should be avoided as it may lead to structural damage of the stents. The current angiological statement concerning stent implantation in the femoral artery may be revised if effective measures to inhibit intimal hyperplasia such as brachytherapy or sirolimus coated stents become available. PMID- 12736703 TI - [Stents in the femoral artery: aid or problem?]. AB - Percutaneous transluminal angioplasty (PTA) is a well-established technique in the treatment of symptomatic femoropopliteal artery stenoses and obstructions. It is a method of treatment with excellent initial technical and clinical results. However, the long-term results of PTA alone must be considered as unsatisfactory as recurrences are frequently observed. In the past, femoropopliteal stenting appeared to be a promising tool to prevent recurrence and to increase patency rates, but it did not produce better results than PTA alone. For this reason, alternative therapeutic strategies such as brachytherapy, drug-eluting pharmastents and gene therapy were developed. Several investigators are beginning to explore their potential use. This article reviews the present possibilities of PTA and stenting in the femoropopliteal region and considers potential future concepts. In this context, the current clinical literature is discussed. PMID- 12736704 TI - [The femoral artery: source for a worst case scenario? Possible fatal outcome succeeding the new DRG-system?]. AB - Stenoses and occlusions of the femoral artery mainly cause claudication. Their course is often silent and patients notice it by chance or in a medical checkup. Mayor amputations are rare in isolated femoral appearance of PAOD (peripheral arterial occlusive disease). Although conservative treatment is very effective in these patients, there are still a lot of operations performed, mainly bypasses, interventional dilatations or stenting. The examples of some typical courses out of more than 64,000 vascular disease patients treated in the Hochrheinklinik during the last 24 years point out a new possibility taking a wrong way in treatment decisions. The structure of the new German DRGs stimulates to carry out interventions and operations as there is a great enhancement in earnings. These may result not only in a rise of costs but in a higher risk for patients, too. PMID- 12736705 TI - [Justification for stents in the femoral artery: a surgical point of view]. AB - Increasingly, endovascular therapy of the superficial femoral artery (SFA) is performed using stent technology. Not only short stenoses, but also longer lesions are receiving primary endovascular treatment, although several randomized studies have shown that stenting the SFA does not improve the prognosis after PTA of lesions of this size. Rather, the stent is indicated as a secondary measure to preserve the PTA-result should complications such as a dissection occur. New technical developments such as nitinol stents, sirolimus or PTFE coated stents offer the prospect of treating more complex SFA lesions (TASC Typ C). However, randomized studies reporting long term results with such stents have yet to be published and any cost-benefit analysis of stent therapy is questionable due to lack of pertinent data. Adjunctive techniques such as laser or brachytherapy have not shown convincing improvement of endovascular SFA therapy. Compound intervention carries the risk of injury to the branching profunda or the popliteal segment, which can transform an otherwise relatively harmless SFA occlusion into a lesion which may endanger the extremity. Furthermore, increasing complexity of the SFA lesion or lesions in diabetic patients result in markedly worse results when stented, in contrast to treatment employing bypass surgery. Applying evidence based criteria to treatment recommendation shows that primary stent-PTA of the SFA is, in most cases, medically and economically unjustifiable. PMID- 12736706 TI - [Chromogenic substrate as antidote against the thrombin inhibitor Melagatran?]. AB - It could be shown in vitro that a chromogenic substrate (Chromozym TH, Roche Mannheim) acts at least partially as antidote against the new thrombin inhibitor Melagatran (AstraZeneca, Molndal, Sweden). It is discussed that this antidote effect of a chromogenic substrate might be due to a substrate competition of fibrinogen, thrombin inhibitor, and chromogenic substrate for thrombin. Further animal experiments will clarify whether this in vitro observation is of practical relevance in vivo, too. PMID- 12736707 TI - CLDN23 gene, frequently down-regulated in intestinal-type gastric cancer, is a novel member of CLAUDIN gene family. AB - Microarray analyses combined with laser-capture microdissection have been applied for risk assessments of gastric cancer as well as for identification of novel genes associated with gastric cancer. EST AA393089 derived from an unknown gene has been reported to be frequently down-regulated in intestinal-type gastric cancer. Here, we identified and characterized the gene corresponding to EST AA393089 by using bioinformatics. EST AA393089 overlapped with BC016047 cDNA, and BC016047 overlapped with EST BM821052. Because the mRNA determined by assembling BM821052 and BC016047 was derived from a novel Claudin (CLDN) family gene, the gene corresponding to EST AA393089 was designated CLDN23. Human CLDN23 mRNA was expressed in germinal center B cells, placenta, stomach as well as in colon tumor. Mouse AK009330 and AK037108 cDNAs were derived from mouse Cldn23 gene. Human CLDN23 (292 aa) and mouse Cldn23 (296 aa) were four-transmembrane proteins, showing 79.5% total-amino-acid identity. WWCC motif, defined by W-X(17-22)-W-X(2) C-X(8-10)-C, was conserved among four-transmembrane proteins of CLDN family. CLDN23 gene, linked to MFHAS1 and PPP1R3B genes, was mapped to human chromosome 8p23.1. CLDN21, CLDN22, and CLDN24 genes were also identified in this study. CLDN21 and CLDN22 genes were located within human genomic contig NT_022792.13. CLDN24 gene on human chromosome 11q23 was located within human genomic contig NT_033899.3. Among 23 CLDN family genes within the human genome, CLDN1 and CLDN16 genes were clustered on human chromosome 3q28, CLDN3 and CLDN4 on 7q11, CLDN6 and CLDN9 on 16p13.3, CLDN8 and CLDN17 on 21q22.11, CLDN21 and CLDN22 on 4q35.1. This is the first report on comprehensive characterization of CLDN23 gene, a candidate tumor suppressor gene implicated in intestinal-type gastric cancer. PMID- 12736708 TI - Assessment of the biological activity of an improved naked-DNA vector for angiogenesis gene therapy on a novel non-mammalian model. AB - From the basic expression vector p/hVEGF165, containing a cDNA sequence encoding the 165-amino-acid isoform of human vascular endothelial growth factor (VEGF165), we generated an improved construct (p163/hVEGF165) by subcloning at the 5' exact end of the VEGF165 cDNA a 163-bp IRES element belonging to the 1,014-bp, 5' untranslated region of the murine VEGF gene. This IRES structure caused enhanced synthesis and increased secretion of the mature protein both in HEK-293 and in COS-7 cells, when compared to the basic construct. Both p/hVEGF165 and p163/hVEGF165 vectors were tested for in vivo angiogenic activity on a novel hirudinean model. As expected, the p/hVEGF165 vector injected as naked DNA was able to induce angiogenesis in the non-vascularized muscular tissue of Hirudo medicinalis. This model also allowed us to monitor intracellular synthesis of VEGF165 and subsequent interstitial secretion from muscle cells. Interestingly, significantly larger muscle tissue areas underwent marked angiogenesis when the improved vector p163/hVEGF165 was injected in H. medicinalis. It thus appears that the p163/hVEGF165 construct allows enhanced expression of the human VEGF165 gene, which in turn is responsible for increased secretion of biologically active growth factor by transduced cells. Since a naked-DNA vector very similar to p/hVEGF165 was recently found to be very active in humans for treatment of heart and limb ischemia, we suggest that our improved construct might be further tested in view of potential therapeutic applications. PMID- 12736709 TI - The human TruB family of pseudouridine synthase genes, including the Dyskeratosis Congenita 1 gene and the novel member TRUB1. AB - A novel human gene denominated TruB pseudouridine (psi) synthase homolog 1 (E. coli) (approved symbol, TRUB1) has been identified and characterized. Spanning approximately 40 kb on chromosome 10 and including 8 exons, TRUB1 is the first described human ortholog of bacterial TruB/psi55, a gene involved in tRNA pseudouridinilation. TRUB1 gene encodes a 349-amino acid product, with a VFAVHKPKGPTSA box in positions 71-83 corresponding to motif I of the TruB family (probably involved in conserving protein structure). The TruB domain of TRUB1 lies between W104 and I255, and contains another short motif, GGTLDS AARGVLVV, including the highly conserved D residue that characterizes motif II (involved in uridine recognition and in catalytic function of psi synthases). Northern blot analysis revealed that TRUB1 mRNA is widely expressed in various human tissues (especially heart, skeletal muscle and liver). Phylogenetic analysis of the TruB domain revealed another human gene (approved symbol TRUB2) encoding a conserved TruB domain, located on human chromosome 9. Thus, the human TruB family includes at least three members: i.e. DKC1 (previously identified), TRUB1 and TRUB2. The TRUB1 and TRUB2 products could be the hitherto unidentified human tRNA psi synthases. Although TRUB1 is not highly similar to DKC1/dyskerin (whose mutations cause X-linked dyskeratosis congenita) and putatively affects tRNA rather than rRNA modification, it is the most similar human protein to dyskerin. Study of TRUB1 (and TRUB2) should facilitate understanding of the molecular mechanisms of RNA modification and the involvement of psi synthases in human pathology, including dyskeratosis-like diseases. PMID- 12736710 TI - Regulation of CREB-mediated transcription by association of CDK4 binding protein p34SEI-1 with CBP. AB - CREB binding protein (CBP) plays a central role in cell differentiation and proliferation, interacting with a large number of nuclear factors. To find novel nuclear factors associating with CBP, we have carried out yeast two-hybrid screening of human chondrocyte cDNA library using the C/H3 region of CBP as a bait and cloned CDK4 binding protein p34SEI-1, the recently found cell cycle regulator. The association of p34SEI-1 with CBP was confirmed in vitro by GST pull-down assay and in vivo by coimmunoprecipitation. Results of the immunofluorescence assay also supported the association of p34SEI-1 and CBP. In reporter assay using CRE promoter, p34SEI-1 strongly suppressed CREB-mediated transcription, and this suppression was overcome by excess amount of CBP, but not by CBPDeltaCH3. It is suggested that the association of p34SEI-1 and CBP is not only involved in cell cycle regulation by CBP, but also have some effect on other CBP-dependent transcription. PMID- 12736711 TI - Profiling of genes differentially expressed between fetal liver and postnatal liver using high-density oligonucleotide DNA array. AB - The liver is an essential organ in humans not only for the production and storage of energy but also for detoxification of chemical compounds, but knowledge about changes in the gene expression profile in the human liver during the prenatal and postnatal periods is limited. Profiling of genes differentially expressed between the fetal liver (FL) and the postnatal liver (PNL) is one of the methods to investigate candidates affecting the difference in biological characteristics between FL and PNL. To identify genes differentially expressed between FL and PNL (childhood and adult liver), we analyzed the gene expression profiles across 9 FL and 14 PNL samples using a high-density oligonucleotide DNA array. Using Mann Whitney U test followed by k-nearest-neighbors (supervised learning method) and hierarchical clustering (unsupervised learning method) algorithms, we found 33 genes clearly discriminating between the FL group and PNL group. The functional classification of the 33 genes identified was related to several kinds of biological pathways, regulating the cell cycle (PCNA, CDC7L1, CCND3, YWHA1, PKMYT1), DNA replication and repair (RFC4, RECQ2, PCNA, NAP1L1), cell growth (IGF2, IGFBP2, PRSS11), hormonal signals (AR, SRD5A1, NR1I3), and cellular metabolism (E2-EPF, WWP1, CYP2C9, CYP2E1, CYP2A6, CYP2A7, CYP2A13, CYP4F2, CYP3A4, DDT). The results presented herein provide evidence of a differential expression profile of genes regulating the cell cycle, DNA replication and repair, cell growth, regulation of hormonal signals, and cellular metabolism, between FL and PNL in humans. The 33 genes identified in this study are suggested to be useful markers clearly discriminating between FL and PNL using the gene expression profile. PMID- 12736712 TI - Late-onset obesity in mice transgenic for the human renin gene. AB - We previously generated a strain of transgenic mice carrying the human renin gene, hRN8-12, in the background of C57BL/6j. In this study, we discovered that hRN8-12 male mice, but not females, developed obesity starting at 15 weeks of age. The body weight of 60-week-old male transgenic mice was 2 times higher than that of age-matched wild-type mice. Interestingly, male mice heterozygous for the human renin gene showed moderate weight gain compared with transgenic and wild type mice. Obese hRN8-12 mice exhibited hyperglycemia, hyperinsulinemia, hyperleptinemia, and hyperlipidemia, and increase in weight in the adipose tissue, liver, heart, and kidneys. Histological analysis demonstrated that fatty hRN8-12 mice developed hypertrophy of pancreatic islets and fatty liver. These results suggested that hRN8-12 mice are associated with obesity dependent on the transgene dosage and should be a genetic model for late-onset obesity. PMID- 12736713 TI - A pilot study of eicosapentaenoic acid therapy for ribavirin-related anemia in patients with chronic hepatitis C. AB - One of the major side effects of ribavirin/interferon alpha combination therapy for chronic hepatitis C is hemolytic anemia. One of the causes of hemolytic anemia is considered to be decreasing deformability of erythrocytes resulting from the accumulation of phosphorylated ribavirin in erythrocytes. The administration of eicosapentaenoic acid (EPA), which has a wide variety of pharmacological actions, increases the deformability of erythrocytes. We conducted an uncontrolled pilot study of EPA therapy for patients with ribavirin related anemia. Six patients with chronic hepatitis C, who had developed anemia while receiving combination therapy, were treated with an oral ethyl ester of EPA (1800 mg/day) for two months. The hemoglobin level of all six patients increased following EPA therapy. The mean hemoglobin level significantly increased from 10.8 g/dl to 11.4 g/dl one month after therapy was initiated (P<0.05), and this level was obtained again one month later (11.5 g/dl). None of the patients developed an adverse reaction. These findings suggest that EPA has a beneficial effect in patients with ribavirin-related anemia. Further study is required to confirm our results. PMID- 12736714 TI - Inhibition of cyclooxygenase-2 and activation of peroxisome proliferator activated receptor-gamma synergistically induces apoptosis and inhibits growth of human breast cancer cells. AB - Cyclooxygenase-2 (COX-2) expression and peroxisome proliferator-activated receptor-gamma (PPARgamma) inactivation are linked to increased risk of human breast cancer. This study examines the effect of simultaneous targeting of COX-2 and PPARgamma on the proliferation of human breast cancer cells and on the expression of Bcl-2, BAX, and caspases-3 and -9, modulators of apoptotic cell death. Treatment of MDA-MB-231 breast cancer cells with NS-398 (a COX-2 inhibitor) or ciglitazone (CGZ, a PPARgamma-ligand) significantly inhibited cell proliferation and markedly increased apoptotic rates. These effects were accompanied by upregulation of BAX and caspases-3 and -9 mRNA expression and downregulation of Bcl-2. Compared to the influence of separate treatments, simultaneous treatment with NS-398 and CGZ synergistically inhibited cell proliferation and induced apoptotic cell death. In conclusion, combinational targeting of COX-2 and PPARgamma can inhibit the growth of human breast cancer cells and induce apoptosis to an extent more suprior to that produced by targeting each molecule alone. COX-2 and PPARgamma can be promising molecular targets for combinational chemoprevention or treatment of breast cancer. PMID- 12736715 TI - Effect of apoptogenic stimuli on colon carcinoma cell lines with a different c myc expression level. AB - We have recently demonstrated that a high c-myc endogenous amplification level confers an apoptosis-prone phenotype to serum-deprived colon carcinoma SW613-S cells. The aim of this study was to gain new insights into the features of c-myc dependent apoptosis, by extending our analysis to different apoptogenic stimuli. The study was carried out on clones, derived from the human colon carcinoma SW613 S cell line, which harbor different levels of endogenous c-myc amplification, and on isogenic cell lines with an enforced c-myc overexpression. Our results indicate that cells with endogenous or transfected exogenous c-myc overexpression (SW613-12A1 and -2G1mycP2Tu1 cell lines, respectively), activate the apoptotic machinery in response to the treatment with etoposide, doxorubicin and vitamin D3, which induce apoptosis through the death receptor Fas. The low levels of c myc expression present in SW613-B3 and -B3mycC5, seem to be unable to activate Fas-mediated apoptosis, thus suggesting that only a high c-myc expression can bypass the lack of Fas receptor. Apoptosis induction mediated by DNA damage and long-term culture was independent of c-myc expression. A pathway of apoptosis characterized by the activation of the enzyme L-DNase II, was observed in both 12A1 and B3 cell lines. PMID- 12736716 TI - The mechanism of 3'-azido-2',3'-dideoxythymidine resistance to human lymphoid cells. AB - The human T-lymphoid cell line H9 resistant to 3'-azido-2',3'-dideoxythymidine (AZT) has a very low level of thymidine kinase (TK) expression which accounts for the failure of AZT to inhibit HIV-1 replication. In the present study DNA methylation and histone deacetylation as possible mechanisms of decreased TK gene expression in the resistant cells were investigated. The resistant cells expressed high levels of DNA methyltransferases (DNMTs) 3a and 3b. The DNA methylation inhibitor, 5-aza-cytidine (5-aza-C), increased TK gene expression and antiviral activity of AZT in the resistant cells, while histone deacetylase inhibitor trichostatin A (TSA) had no effect. The results suggest that hypermethylation of the TK gene but not histone deacetylation in AZT-resistant H9 cells accounts for decreased TK gene expression and failure of AZT to inhibit HIV 1 replication probably due to overexpression of DNMT 3a and 3b. PMID- 12736717 TI - Predictive factors for remission and death in 73 patients with autoimmune hepatitis in Japan. AB - The prognosis of patients with autoimmune hepatitis (AIH) has not been clearly defined. The aim of this study was to define the prognostic factors of AIH in a population with long-term follow-up in Japan. Seventy-three patients who were diagnosed as having type 1 AIH between January, 1972 - August, 1999 were enrolled in this study. Initial treatment included prednisolone (PSL) (n=62), other drug regimens (n=7), and none (n=4). We examined the relation between several factors obtained at diagnosis in relation to disease activity found at the final observation point (January, 2000 - April, 2000). Multivariate logistic regression and Cox regression were used for statistical analysis. During the observation period, 8 patients died of the following: hepatic failure (n=4), hepatocellular carcinoma (n=1), severe infection (n=1), and unknown causes (n=2). At the end point, the number of patients in complete remission was 13, those with a normal alanine aminotransferase (ALT) level requiring some treatment was 35, and those with an abnormal ALT level despite medication was 17. Factors related to remission were total bilirubin (TB) (Odds ratio, 0.87), and immunoglobulin G (IgG) (Odds ratio, 1.00). Factors related to death were the aspartate aminotransaminase (AST)/ALT ratio (Odds ratio, 11.67) and response to initial PSL regimen (Odds ratio, 0.03). The results of this study show an importance of achieving a good PSL response at onset, and that initial TB, the AST/ALT ratio, and IgG levels are useful for therapeutic strategy. PMID- 12736718 TI - Fas antigen (CD95) mediates cell survival signals to regulate functional cellular subpopulations in normal human endometrial stromal cells. AB - Fas antigen (CD95) is expressed on almost all types of human cells and is believed to mediate receptor-specific apoptotic signals. In human endometrial tissues, high Fas and Fas ligand expressions and Fas-mediated apoptosis in endometrial epithelial cells have been discussed in many reports but no study has examined Fas-mediated signals in endometrial stromal cells. In this study we investigated Fas expression and Fas-mediated signals of normal human endometrial stromal cells. Flow cytometric analysis revealed Fas antigen expression on the stromal cells and their Fas expression was enhanced by 8-Br-cAMP, a strong inducer of decidualization. Neither short-term nor long-term cultures with anti Fas IgM affected proliferation or viability of the stromal cells. Anti-Fas IgM alone affected neither viable cell numbers nor PRL release of unstimulated stromal cells. However, anti-Fas IgM dose-dependently stimulated viable cell numbers of stromal cells co-stimulated with 8-Br-cAMP and anti-Fas IgM, whereas PRL secretion of the co-stimulated cells was not affected. Anti-Fas IgM dose dependently stimulated viable cell numbers of 8-Br-cAMP-stimulated cells but did not affect PRL secretion of 8-Br-cAMP-induced decidualized cells. These results indicate that Fas antigen on human endometrial stromal cells cannot mediate receptor-specific apoptotic signals, and that Fas-mediated signals stimulate survival of 8-Br-cAMP-stimulated non-decidualized stromal cells. Thus, stimulation of Fas-antigen on the endometrial stromal cells enhances anti apoptotic/survival signals in certain stromal cells, autoregulating functional cellular subpopulations of human endometrial stromal cells in a paracrine manner. PMID- 12736719 TI - Effects of prolonged exendin-4 administration on entero-insular axis of normal and streptozotocin-induced diabetic rats. AB - The effects of the glucagon-like peptide 1 (GLP-1) receptor agonist exendin-4 (EX4) and antagonist EX4(9-39) EX4-A on entero-insular axis have been investigated in normoglycemic and streptozotocin (STZ)-induced diabetic rats. Rats were administered daily subcutaneous injections of 1 nmol/kg EX4 and/or EX4 A for 7 days, and were decapitated 3 h after the last injection. In STZ-untreated rats, EX4 reduced body-weight (BW) gain and raised glycemia, and the effects were prevented by EX4-A; conversely, EX4 did not alter plasma concentrations of insulin, glucagon and leptin. STZ-treated rats displayed body and hematochemical alterations typical of experimental diabetes: decrease in BW and insulin blood level, coupled with normal glucagon plasma concentration and marked hyperglycemia. In diabetic rats, both EX4 and EX4-A decreased BW gain, thereby suggesting a mechanism at least in part independent of GLP-1 receptors. EX4 did not alter glucagon blood level, but decreased glycemia and raised insulin and leptin plasma levels. These effects were annulled by EX4-A, which indicates that they occur through the activation of GLP-1 receptors. Collectively, our findings add support to the view that EX4 can be considered an important therapeutical tool to improve glucose metabolism in diabetes. PMID- 12736720 TI - Manumycin inhibits cell proliferation and the Ras signal transduction pathway in human hepatocellular carcinoma cells. AB - Manumycin was reported to have inhibitory effect on farnesyltransferase by competing with the farnesyl pyrophosphate substrate. It exhibited different antiproliferative activity in human hepatocellular carcinoma HepG2 cells, primary cultured human cardiac muscle cells and human liver cells (CLC). HepG2 cells overexpressing ras gene were more sensitive to manumycin than the other cells. The difference might be related to Ras protein levels in these cell lines. Manumycin reduced the amount of functional ras localized at the cytoplasmic membrane, resulting in blocked C-raf-1 assocation with Ras. Manumycin inhibited ERK1/2 phosphorylation in HepG2 cells without reduced expression of ERK1/2 protein. The levels of protein MKP-1 were significantly up-regulated. Our study also demonstrated that manumycin inhibited p85/PI3K and Akt phosphorylation without reduced expression of p85/PI3K and Akt, and interfered with the association of p85/PI3K and Ras. These findings indicated that manumycin interfered with Ras membrane localization, shut down the downstream pathways of Ras and inhibited cell proliferation in HepG2 cells. PMID- 12736721 TI - Genetic changes and expression of the mannose 6-phosphate/insulin-like growth factor II receptor gene in human hepatitis B virus-associated hepatocellular carcinoma. AB - It was reported that 60-70% of hepatitis B virus (HBV)-negative hepatocellular carcinoma (HCC) had loss of heterozygosity (LOH) at the mannose 6 phosphate/insulin-like growth factor II receptor (M6P/IGF2R) locus and this gene was mutated in 55% of these patients with LOH. In this study, genomic DNA from 29 pairs of HBV-positive HCC and corresponding non-tumor tissues was used to analyze LOH at the M6P/IGF2R locus and single deoxyguanosine deletion in this gene by PCR. Total RNA from 19 of the 29 patients was utilized to determine a 192 bp insert in the M6P/IGF2R mRNA and expression of this gene by RT-PCR. Twenty-eight of 29 (97%) HBV-positive HCC were found to be informative at the M6P/IGF2R locus but LOH at this region was only detected in 4/28 (14%) informative patients. Neither single deoxyguanosine deletion in this gene nor 192 bp insert in its mRNA occurred in these patients. Compared with corresponding non-tumor tissues, expression of the M6P/IGF2R mRNA was decreased in 13/19 (68%) HBV-positive HCC tissues, suggesting that M6P/IGF2R may be involved in HBV-associated hepatocarcinogenesis by the regulation of its expression level. In the development of HBV-associated HCC, M6P/IGF2R mutation may not be a major agent. PMID- 12736722 TI - Quantification of human cytomegalovirus using bronchoalveolar lavage cells in pulmonary complications associated with hematologic neoplasia. AB - Human cytomegalovirus (CMV) has been recognized as a frequent pathogen involved in interstitial pneumonia (IP), and CMV-IP is a severe and life-threatening complication in the immunocompromised patients. The use of real-time PCR in molecular diagnostics has increased to the point where it is now accepted as the gold standard for detecting a wide variety of templates including viruses. Therefore, we developed a rapid quantification system of CMV using a LightCycler in order to clarify the possible role of CMV reactivation in patients with hematologic neoplasia showing pulmonary complications. Sixty-nine bronchoalveolar lavage fluid (BALF) specimens were obtained from consecutively treated patients showing interstitial shadow including 20 patients with hematologic neoplasia. First, we determined the viral burden in BAL cells from healthy volunteers, idiopathic interstitial pneumonia (IIP) and sarcoidosis. CMV copy numbers in samples obtained from healthy volunteers, IIP and sarcoidosis, were less than 10(2) copies per 1 microg of DNA, whether or not BAL cells were composed of high percentage of lymphocytes. Among 20 patients with hematologic neoplasia analyzed, two specimens obtained from leukemia patients with pulmonary alveolar proteinosis, two from GvHD, one with CMV interstitial pneumonia and one with Hodgkin's disease had high level of CMV viral DNA. Our results suggest that measurement of CMV genomes in BAL cells using real-time PCR may be useful not only to understand the involvement of CMV in systematic respiratory tract disease but also in management of the care of respiratory complications in hematologic neoplasia. PMID- 12736723 TI - Effects of thalidomide on the expression of angiogenesis growth factors in human A549 lung adenocarcinoma cells. AB - Angiogenesis is a critical step for the growth and metastasis of solid tumors, and it is a major therapeutic target for the development of anti-angiogenics for cancer treatment. Thalidomide is reported to be an anti-angiogenic agent, which is currently in phase II clinical trials for the treatment of advanced malignancies. However, the mechanism of thalidomide in angiogenesis is not completely understood. This study was undertaken to investigate the effect of thalidomide on the expression of angiogenesis growth factors in human lung cancer cells. In this report, we show that thalidomide downregulated the mRNA and protein expression of basic fibroblast growth factor (bFGF) in the human lung adenocarcinoma cell line A549, and that the effect of thalidomide was time and concentration-dependent. In contrast, the expression of vascular endothelial growth factor (VEGF) by thalidomide in these cells was in two phases. The mRNA and protein expression of VEGF was increased in the lung cancer cells treated with 0.6-6 microg/ml thalidomide, while higher concentrations of thalidomide decreased VEGF levels significantly in these cells. These data suggest that the anti-angiogenic or antitumor activity of thalidomide may be partly mediated through the regulation of the levels of angiogenesis growth factors. PMID- 12736724 TI - FNBP2 gene on human chromosome 1q32.1 encodes ARHGAP family protein with FCH, FBH, RhoGAP and SH3 domains. AB - Parallel analyses of DNA copy number and mRNA expression level by using microarray measurements have been successfully applied for genome-wide screening of proto-oncogenes and tumor suppressor genes. The uncharacterized KIAA0456 cDNA was reported amplified and overexpressed in human breast cancer cell lines UACC812 and ZR-75-1. Here, we characterized the gene corresponding to KIAA0456 cDNA by using bioinformatics. KIAA0456 cDNA was found derived from the FNBP2 gene, consisting of 22 exons. FNBP2 gene was linked to IKBKE and NORE1 genes on human chromosome 1q32.1. FNBP2 mRNA was expressed in melanoma, germ cell tumors, chondrosarcoma and retinoblastoma. ARHGAP13/SRGAP1, ARHGAP14/SRGAP2 and ARHGAP4 cDNAs were homologous to FNBP2/KIAA0456 cDNA. KIAA1304 cDNA was a splicing variant derived from the ARHGAP13 gene, and the nucleotide sequence of representative ARHGAP13 cDNA was determined by assembling EST BU520980 and KIAA1304 cDNA. FNBP2 (1071 aa), ARHGAP13 (1062 aa), ARHGAP14 (1099 aa) and ARHGAP4 (946 aa) constitute the FNBP2 family characterized by FCH, RhoGAP and SH3 domains. The region corresponding to codon 227-345 of FNBP2 was conserved among FNBP2 family proteins as well as FNBP1 and TRIP10 proteins. Because FNBP2 and FNBP1 are formin-binding proteins, the region corresponding to codon 227-345 of FNBP2 was designated FNBP2-FNBP1 homologous (FBH) domain. FNBP2 family proteins consist of FCH, FBH, RhoGAP and SH3 domains, while FNBP1 family proteins (FNBP1 and TRIP10) consist of FCH, FBH and SH3 domains. This is the first report on comprehensive characterization of FNBP2 gene as well as on identification of the FBH domain. PMID- 12736725 TI - Effect of Bcl-2 antisense oligonucleotide on drug-sensitivity in association with apoptosis in undifferentiated thyroid carcinoma. AB - Although attempts have been made to treat undifferentiated thyroid carcinoma using multidisciplinary therapeutic procedures including surgery, radiotherapy, and chemotherapy, the prognosis of undifferentiated thyroid carcinoma remains quite poor. New approaches to increase the sensitivity of patients to anticancer drugs and radiation will be needed to improve the survival rate for undifferentiated thyroid carcinoma. We examined the effect of Bcl-2 antisense oligonucleotide on drug-sensitivity in association with apoptosis in the 8305C undifferentiated thyroid carcinoma cell line. The drug sensitivity was evaluated by MTT assay for 48 h, while apoptosis was assessed according to the formation of internucleosomal DNA ladders. The Bcl-2 antisense was introduced into 8305C cells by using a 18-mer phosphorothioate oligonucleotide by lipopolyamine-mediated transfection twice for 12 h. The expression of apoptosis genes was assessed by Western blotting. The 8305C cells were sensitive to adriamycin (ADM), mitomycin (MMC), docetaxel (TXT), and paclitaxel (TXL), showing mean IC50 values of 0.72, 1.1, 1.3, and 4.1 microM, respectively. In contrast, the 8305C cells were resistant to cisplatin (CDDP) and 5-fluorouracil (5-FU), with mean IC50 values of 42.0 and 48.0 microM, respectively. Treatment with Bcl-2 antisense suppressed the protein level of Bcl-2 in 8305C cells in a dose-dependent manner up to 1.0 microM. Drug-sensitivity was increased by pretreatment with Bcl-2 antisense as assessed by the IC50 (x-fold): 0.48 (1.5-fold) in ADM; 0.42 (2.6-fold) in MMC, 0.56 (2.3-fold) in TXT, 1.5 (2.7-fold) in TXL, 8.6 (4.9-fold) in CDDP, and 25.0 (1.9-fold) in 5-FU, respectively. The increased drug-sensitivity was associated with the induction of apoptosis-related proteins, Fas, caspase 8, cytochrome c, caspase 3, and to subsequent apoptosis, as determined by the formation of internucleosomal DNA ladders and PARP in the treated cells. Susceptibility in apoptotic cell death following treatment with anticancer drugs was associated with induction of apoptosis-related genes in undifferentiated thyroid carcinoma cells, and induction of apoptosis was enhanced by pretreatment with Bcl-2 antisense oligonucleotide. These results imply a potential new strategy targeting an antiapoptotic protein, Bcl-2, by its antisense oligonucleotide for enhancement of chemotherapeutic efficacy in undifferentiated thyroid carcinomas. PMID- 12736726 TI - The expression of platelet endothelial cell adhesion molecule-1 in mouse primordial germ cells during their migration and early gonadal formation. AB - The platelet endothelial cell adhesion molecule-1 (PECAM-1), or CD31, a member of the immunoglobulin superfamily, is located on the plasma membrane of endothelial and hematopoietic cells and involved in vascular development and inflammation. In this study, by use of immunohistochemistry at light and electron microscopic levels in combination with enzyme histochemistry for alkaline phosphatase, we demonstrated that PECAM-1/CD31 is expressed in the mouse primordial germ cell (PGC). Up to 8 days postcoitum (dpc), PGCs with alkaline phosphatase activity showed no PECAM-1/CD31 immunoreactivity. At 9 dpc, PECAM-1/CD31 immunoreactivity was first detected with low intensity in some PGCs located in the hindgut. Between 10 and 11 dpc, intense immunoreactivity was shown on the entire surface of PGCs migrating along the dorsal wall. After arrival and settlement of PGCs in the genital ridges around 11.5 dpc, the intense immunoreactivity was maintained on the entire surface of PGCs. By electron microscopy, the immunoreactivity was localized exclusively on the plasma membrane of PGCs, being as strong at the portions adjacent to neighboring PGCs as those adjacent to somatic cells. As the male and female gonads began to differentiate, PECAM-1/CD31 immunoreactivity remained strong in germ cells until 13 dpc, after which it gradually decreased in intensity and disappeared by 16 dpc. These results suggested that cell-to-cell interaction through PECAM-1/CD31 plays roles in the development of PGCs during their migration on the dorsal wall and homing in the gonads. PMID- 12736727 TI - Clinical findings in macular hole surgery with indocyanine green-assisted peeling of the internal limiting membrane. AB - PURPOSE: Indocyanine green (ICG) staining of the internal limiting membrane has facilitated ILM peeling in macular hole surgery. However, it has been reported that ICG-assisted peeling of the ILM may result in retinal damage and unfavorable functional outcome. Therefore, we analyzed our visual and anatomical results of ICG assisted macular hole surgery. METHODS: In a retrospective study the records of a consecutive series of 37 patients with full-thickness idiopathic macular holes operated with ICG-assisted ILM peeling by a single surgeon were analyzed. All patients underwent a standard three-port vitrectomy with surgically induced posterior vitreous detachment, staining of the ILM with ICG, peeling of the ILM in a circular manner around the fovea, and SF6 20% endotamponade. RESULTS: A total of 37 patients (37 eyes) were included in the study. The mean age was 69+/ 7 years (range 52-81 years), and there were 26 women and 11 men. The follow-up ranged from 6 to 30 months (mean 18+/-6 months). At baseline visual acuity ranged from 20/400 to 20/40. Anatomically, 13 eyes had stage 2 holes, 21 eyes (57%) stage 3 holes, and three eyes stage 4 holes. At the postoperative visit (8-12 weeks after surgery) anatomical closure of the macular hole was achieved in 36 eyes. Visual acuity ranged between 20/400 and 20/20. At the last follow-up after initial surgery the macular hole was closed in all eyes. Visual acuity ranged from 20/200 to 20/20. CONCLUSION: In our retrospective series anatomical and functional results of macular hole surgery with ICG-assisted peeling of the ILM are satisfactory. Primary hole closure was achieved in 97% of eyes and visual acuity increased in 62% of eyes in our series. PMID- 12736728 TI - Refractive changes in pregnancy. AB - OBJECTIVE: To determine the causes of any vision change reported during pregnancy. SETTING: An obstetrical practice in Southampton, New York. STUDY POPULATION: Two hundred forty pregnant women were asked whether they had any alteration in vision. Those who agreed to take part in the study (83) and who complained of vision changes (12) were matched with the next patient seen in the practice who was asymptomatic. OBSERVATION. All patients underwent a complete ophthalmic examination, including refraction. Those who had alterations in vision status were seen again after delivery. MAIN OUTCOME MEASURES: Changes in visual acuity and refractive error during pregnancy. RESULTS: All women who complained of visual changes were found to have experienced a myopic shift from pre pregnancy levels. (0.87+/-0.3 diopters in the right eye ( P<0.0001) and 0.98+/ 0.3 diopters in the left eye ( P<0.0001). Post partum, all subjects returned to near pre-pregnancy levels of myopia. CONCLUSIONS: This report links worsening of myopia to pregnancy. The causes of this myopic shift are not readily evident and merit further investigation. PMID- 12736729 TI - Dorsolateral prefrontal lobe activation declines significantly with age- functional NIRS study. AB - Dorsolateral prefrontal cortex (DLPF) activation in response to cognitive paradigms engaging working memory was quantitatively evaluated using functional near infrared spectroscopy (fNIRS) in cognitively normal individuals across the age spectrum (20-90 years; 8 females, 52 males). DLPF activation score (DAS) demonstrated a significant decline (r(2) = -0561, p < 0.05) as a function of age. The study indicates that blood flow response associated with DLPF activation declines as a function of age in cognitively normal individuals and that fNIRS can be used as a convenient, portable tool for assessing such activation. PMID- 12736730 TI - Autonomic dysfunctions in dementia with Lewy bodies. AB - Twenty-nine cases of both clinically and neuropathologically diagnosed dementia with Lewy bodies (DLB) were retrospectively examined for autonomic symptoms. Twenty-eight cases showed some kind of autonomic dysfunction. Urinary incontinence (97 %) and constipation (83 %) were the two most common. Although urinary retention and episodic hypotension causing syncopal attacks were less common, the frequency was still high (28 % each). There were 18 cases (62 %) with severe autonomic failure. These 28 cases showed similar tendencies, with no significant differences between the subtypes of DLB (brainstem, limbic, and neocortical types or common and pure forms). We found that DLB of all pathological subtypes exhibits some kind and level of autonomic symptoms. PMID- 12736732 TI - Misoprostol in the treatment of trigeminal neuralgia associated with multiple sclerosis. AB - Multiple sclerosis can be associated with trigeminal neuralgia which is often difficult to treat in this specific condition. We performed an open prospective trial on the efficacy and safety of the prostaglandin-E1-analogue misoprostol (600 microg per day) in the reduction of attack frequency and pain intensity in patients with refractory trigeminal neuralgia associated with multiple sclerosis. Eighteen patients completed the study period and 14 of them showed a reduction of more than 50 % in attack frequency and intensity beginning five days after treatment onset. There were only mild and transient drug related side effects in three patients. One patient stopped taking misoprostol after the study period because of severe menorrhagia. Our results suggest that misoprostol is effective and safe in the treatment of this specific type of refractory trigeminal neuralgia. PMID- 12736731 TI - Clinimetric evaluation of a Satisfaction-with-Stroke-Care questionnaire. AB - There is a lack of sound instruments for measuring patient satisfaction with stroke care. One self-report instrument comprising two subscales, satisfaction with inpatient care and satisfaction with care after discharge, has been validated, but only in the United Kingdom. In later studies, items have been added without further validation. Therefore, we tested this extended questionnaire (Satisfaction with Stroke Care questionnaire; SASC-19) for feasibility, reliability (homogeneity and test-retest agreement), and construct validity (convergent and divergent validity and factor analysis) in the Netherlands in 166 prospectively collected stroke patients living at home 6 months after discharge. To determine the test-retest reliability, 51 patients completed the SASC-19 again two weeks after they first completed it. The response rate was 90 %; the mean time needed to complete the SASC-19 was less than 10 minutes. Six items were omitted by more than 10 % of the patients. Both subscales showed good homogeneity and almost perfect test-retest reliability (Cronbach's alpha's > 0.80; Intraclass Correlation Coefficients > 0.80). The correlations with the General Satisfaction questionnaire (convergent validity) ranged between 0.33 and 0.55; those with the Barthel Index, the Hospital Anxiety and Depression Scale (HADS), and the Short Form 36 (SF-36) health survey questionnaire (divergent validity) ranged between 0.12 and 0.47. Factor analysis showed a total explained variance of 49 %, which supports the subscale structure. We conclude that the SASC-19 is a reliable and valid instrument for measuring patient satisfaction with stroke care and it is easy to complete. The feasibility in terms of missing values is moderate. PMID- 12736733 TI - Idiopathic spasmodic torticollis is not associated with abnormal kinesthetic perception from neck proprioceptive and vestibular afferences. AB - Proceeding from recent evidence for a sensory involvement in the pathophysiology of idiopathic spasmodic torticollis (ST), we asked whether the abnormal head posture of these patients is associated with distortions of their internal spatial reference frames due to abnormal processing of neck proprioceptive and/or vestibular input. Twelve ST patients were instructed to estimate, by adjusting a light pointer in the dark, their head and trunk mid-sagittal directions (as representatives of ego-centric references) and to reproduce a remembered target location in space (space centric reference). They did so before and after horizontal head and trunk rotations, which evoked isolated or combined vestibular and/or neck stimulation. In ST patients, unlike in normal controls, pre-stimulus estimates of the head and trunk mid-sagittal directions (baselines) showed a pronounced across-subjects variability, with essentially normal mean values. Their post-stimulus estimates in all tasks, after correction for the individual baseline errors, were normal with respect to both amplitude and variability, independent of stimulus direction, modality and rotation dynamics. Our findings suggest that ST patients have a rather inaccurate knowledge of their head posture, but can effectively use neck proprioceptive input and vestibular cues when estimating head and trunk displacements in ego-centric and space centric spatial orientation tasks. We propose that an offset of a non-sensory set point signal in the neck proprioceptive loop for head-on-trunk control may be responsible for the pathological head deviation in ST. PMID- 12736734 TI - Poor metabolization of n-hexane in Parkinson's disease. AB - Although genomic screening studies have identified several genes associated with Parkinson's disease (PD), there is evidence that environmental factors are also involved in the pathogenesis of the disease and that hydrocarbon-solvents may be one of them. The genetic component is less evident in late-onset PD. To assess whether age and PD may affect the catabolism of the hydrocarbon n-hexane, a two part study was performed. In the first part the urinary levels of its main metabolites, 2,5-hexanedione and 2,5-dimethylpyrroles, were measured in 108 patients and 108 healthy controls, matched by age and sex. Metabolite urinary excretion was significantly reduced in PD patients as compared with controls and was inversely related to age in both groups. In the second part the same comparison was made between 24 non-smoking and 10 smoking patients, matched to controls, after smoking of a hydrocarbon-rich cigarette. In these subjects also n hexane and 2,5-hexanedione blood levels were measured. There was no appreciable difference in n-hexane blood levels between patients and controls in non-smokers, whereas there was a significant increase in patients over controls in smokers (p < 0.01). 2,5-hexanedione blood levels were significantly lower in patients than in healthy controls, both in non-smokers and in smokers, but the reduction was more pronounced in smokers (-46.3 % versus -10.7 %). The same was true for 2,5 hexanedione and 2,5-dimethylpyrrole urinary levels. This study suggests that aging and PD may be associated with a reduction in the capacity to eliminate the hydrocarbon n-hexane. This metabolic alteration may play a role in the pathogenesis of PD. PMID- 12736735 TI - Quality-adjusted survival after tumor resection and/or radiation therapy for elderly patients with glioblastoma multiforme. AB - OBJECTIVE: Prognostic factors are poorly defined for the elderly subpopulation with glioblastoma multiforme and have been exclusively related to conventional survival analysis. In this study an additional quality adjusted survival analysis (QAS) was performed. The prognostic evaluation of both survival- and QAS data after standard treatment were checked for concordant/discordant findings. Their usefulness for estimation of treatment effects and treatment strategies was then evaluated. METHODS: 123 patients >or= 65 years of age with a supratentorial, de novo glioblastoma were included in the current retrospective report. Microsurgery plus radiation therapy (planned tumor dose: 60 Gy) was performed in 58 patients, and radiation therapy alone after stereotactic biopsy (planned tumor dose: 60 Gy) in 65 patients. The functional status of each patient was scored when joining the study and at every follow-up using 15 selected neurological signs and symptoms (NSSs). Gradation of severity of each NSS was performed with subjective weights. Survival time of each patient was adjusted according to any changes in these NSSs to become the Quality Time (Q-TIME). Time intervals spent with side effects of the treatment (TOX) were subtracted from Q-TIME to become the patient's QAS (QAS = Q-TIME-TOX). Prognostic factors for both survival and QAS were obtained from the Cox model. RESULTS: Overall survival and QAS were 24 weeks and 10.5 weeks, respectively. Perioperative morbidity and mortality were 5.2 % and 1.7 % in the surgery group and 1.5 % and 1.5 % in the biopsy group, respectively (p > 0.05). Tumor resection gained favorable prognostic importance for patients with midline shift in terms of both survival and QAS (p < 0.0001). Otherwise, radiation therapy alone was as effective as surgery plus radiation therapy (concordant finding). A pretreatment Karnofsky Score (KPS) < 70 was an unfavorable predictor for QAS (p < 0.002) but not for survival (discordant finding). Median QAS for patients with a pretreatment KPS < 70 was only 10 weeks. Age did not reach prognostic relevance. CONCLUSION: The dramatic decrease of QAS as compared with survival indicates extremely limited posttreatment improvement and/or rapid deterioration of the neurological score after standard treatment for the older subpopulation with glioblastoma multiforme. Supportive treatment should be considered for patients with a pretreatment KPS < 70. PMID- 12736736 TI - Role of eye movement examination and subjective visual vertical in clinical evaluation of multiple sclerosis. AB - BACKGROUND AND PURPOSE: Modern neuroimaging has demonstrated progression of disease in multiple sclerosis (MS) that may not be detected by standard clinical protocols, prompting a search for new, sensitive tests. METHODS: In fifty patients with MS, we examined eye movements, with particular attention to the speed and accuracy of saccades, and the vestibulo-ocular reflex during small, high-speed head rotations. We also measured the subjective visual vertical (SVV), using a modified laser-pointer. Visual function was measured, and patients were graded using the Kurtzke Functional Neurological Status (FSS), and Expanded Disability Status Scale (EDSS). RESULTS: Our main finding was that patients showing abnormalities of eye movements (20/50) were more disabled than those with a normal examination (EDSS scores significantly different, p < 0.01), although the ages and duration of disease were similar in both groups. Saccadic dysmetria and internuclear ophthalmoparesis were common. SVV was abnormally large in 36 % of patients; these showed abnormal eye movements and poorer visual acuity more commonly than those with normal SVV. In patients with the largest deviations of SVV, Kurtzke FSS cerebellar functions were significantly worse (p = 0.021). CONCLUSIONS: Clinical examination of eye movements, with attention to dynamic properties of saccades and the vestibulo-ocular reflex, takes only a few minutes to perform, but may provide better information concerning the presence of brainstem and cerebellar involvement than Kurtzke protocols. Measurement of SVV is possible in the clinic and is a sensitive sign of brainstem dysfunction; our present study suggests that SVV is also affected when cerebellar circuits are involved in MS. Prospective studies are required to determine whether the development of abnormalities with ocular motor and SVV testing are predictive of disease activity and progressive disability in MS. PMID- 12736737 TI - The degree of depression in Hamilton rating scale is correlated with the density of presynaptic serotonin transporters in 23 patients with Wilson's disease. AB - OBJECTIVE: One of the most frequent psychiatric symptoms in patients with Wilson's disease (WD) is depression. It has been suggested that depression is associated with deficits in serotonergic neurotransmission, but, hitherto, no measurements have been performed in WD. METHODS: We prospectively examined 23 adult patients (12 women, 11 men, mean age 40 years) with WD for symptoms of depression using the Hamilton rating scale for depression (HAMD). We correlated the data with the presynaptic serotonin transporter density (SERT density) in the thalamus-hypothalamus and the midbrain-pons regions measured with high resolution single-photon emission computed tomography (SPECT) 24 hours after the application of 180 MBq 2beta-carbomethoxy-3beta-(4 [(123)I]iodophenyl)tropane ( [(123)I]b CIT). The regions of interest were determined by coregistration with a standard MRI dataset. RESULTS: A significant negative correlation was found between HAMD and SERT density in the thalamus-hypothalamus region (r = -0.49, p = 0.02), but not in the midbrain-pons (r = -0.31, p = 0.15). CONCLUSIONS: We conclude that depression in patients with Wilson's disease is correlated with alterations of serotonergic neurotransmission in the thalamus-hypothalamus region. PMID- 12736738 TI - Analysis of dystrophin mRNA from skeletal muscle but not from lymphocytes led to identification of a novel nonsense mutation in a carrier of Duchenne muscular dystrophy. AB - Dystrophin mRNA expressed in peripheral lymphocytes of individuals with X-linked Duchenne muscular dystrophy (DMD) has been used as a source material for mutation analysis. Here we present the first report of failure of isolation of nonsense dystrophin mRNA in lymphocytes but success in skeletal muscle in a female carrier of DMD. The mutation responsible for dystrophin-negative muscle fibers of the carrier was analysed by direct sequencing of the reverse transcription PCR product of dystrophin mRNA. In her peripheral lymphocytes, no nucleotide change was detected in the 14 kb long mRNA. Remarkably, a novel nucleotide change of C1682T in exon 12, changing glutamine codon to stop codon (Q492X) was found to be present in her skeletal muscle. This change was heterozygous. Analysis of her genomic DNA disclosed heterozygous C and T nucleotides at nt 1682, confirming the genomic origin of the nonsense mutation. Although dystrophin cDNA prepared from lymphocytes was sequenced again after subcloning, mutation-retaining clone could not be isolated. This lymphocyte-specific disappearance of nonsense mRNA strongly suggested tissue-specific skewing of X-inactivation. However, both paternal and maternal dystrophin alleles were shown to be equally expressed in lymphocytes as well as in muscle, indicating no skewing of X-inactivation in lymphocytes. We concluded that the dystrophin mRNA of the DMD carrier was destabilized in lymphocytes. Our results indicated that analysis of mRNA in lymphocytes is not enough for exact carrier diagnosis of Duchenne muscular dystrophy. PMID- 12736739 TI - Nitric oxide as an activity marker in multiple sclerosis. AB - Nitric oxide (NO) molecules have one of the most important roles in the pathogenesis of multiple sclerosis (MS). It has been stated that a continuous and high concentration of NO metabolites in CSF and in the serum of MS patients in relapse may cause toxic damage to myelin and oligodendroglia. The aim of this study was to investigate whether NO is a marker of disease activity and is correlated with other disease activity markers such as active lesions on brain magnetic resonance imaging (MRI) and increased immunoglobulin G (IgG) index. Cerebrospinal fluid (CSF) and peripheral serum (PS) samples were taken from patients with definite MS (n = 24) during relapse and remission and from control subjects (n = 18). The Griess reaction was used to measure the NO metabolites, nitrite and nitrate in CSF and PS. Cranial MRI was carried out with triple dose (0,3 mmol/kg) gadolinium and the IgG index was determined. Nitrite and nitrate concentrations (NNCs) of CSF were 11.16 +/- 8.60 micromol/ml in relapse and 6.72 +/- 3.50 micromol/ml in remission, whereas in PS they were 12.89 +/- 7.62 micromol/ml during relapse and 12.35 +/- 6.62 micromol/ml during remission. In control subjects NNCs in CSF and PS were 7.42 +/- 2.81 micromol/ml and 4.37 +/- 1.63 micromol/ml respectively. NNCs in CSF during relapse period were significantly higher than those of both remission phase and control subjects (p = 0.000). Although serum NNCs did not differ in relapse and remission, they were still higher than normal controls. Validity analysis revealed that NNC measurement in CSF was 71 % specific and 66 % sensitive to disease activity. The most important result was the significant correlation of increased NNCs with the existence of active lesion in cranial MRI and an increase in IgG index (p < 0.05).In conclusion, these results add background data to assist in further outlining the possible role of NO in the pathogenesis of MS. Together with the other markers it may be used as an activity marker in relapses of MS. PMID- 12736740 TI - Selection of thrombogenetic antiphospholipid antibodies in cerebrovascular disease patients. AB - BACKGROUND AND PURPOSE: The association between anticardiolipin antibodies (aCL) and thrombosis is well recognized, but its role as an independent risk factor for stroke is not. The study's aim was to investigate the presence of antiphospholipid antibodies (aPL) and ischemic vascular events by using both traditional means the estimation of aCL and glycoprotein (beta(2)GP1) antibodies. Additionally both aCL/beta(2)GP1 and aPLmix/beta(2)GP1 antibodies were measured. The measurement of these two antibodies was determined by using as target antigens, either cardiolipin alone or a mixture of different phospholipids coated with human beta(2)GP1 in order to select only the autoimmune antibodies. One hundred and twenty-two consecutive patients with first-ever acute ischemic cerebrovascular event were included and compared with controls. The presence of aCl in patients (20.5 %) and controls (14.7 %) was not significantly different (p = 0.1). The presence of abeta(2)GP1 (6.5 % versus 4.9 %, p = 0.7) was also not significant, while there were associations for aCL/b2GP1 13.9 % versus 4.9 % (p = 0.02) and aPLmix/beta(2)GP1 15.6 % versus 4.9 % (p = 0.01). These latter tests seem to be useful in assessing the autoimmune and therefore the thrombogenetic antibodies. PMID- 12736741 TI - No association of inducible nitric oxide synthase gene ( NOS2A) to multiple sclerosis. AB - Nitric oxide (NO) is a nonspecific inflammatory mediator that has been involved in the pathogenesis of multiple sclerosis (MS). The influence of the promoter polymorphism of inducible NO synthase gene ( NOS2A) on susceptibility and outcome was studied in 140 MS Spanish patients and 147 healthy controls matched for sex, age, and ethnicity. No association was found between MS susceptibility, course or outcome of the disease, and NOS2A polymorphisms. PMID- 12736742 TI - Assessment of esophageal function in patients with myasthenia gravis. AB - BACKGROUND: Myasthenia gravis (MG) is an autoimmune disease in which impairment of neuromuscular transmission results in a pathological fatigability of striated muscles. Dysphagia is a common symptom in MG. It is caused by a weakness of the striated muscles in the pharynx and esophagus. The purpose of our study was to evaluate the role of esophageal scintigraphy in the assessment of esophageal function in MG. METHODS: In 15 patients with clinically proven MG (oculopharyngeal manifestation in 6/15 patients, generalized weakness in 9/15 patients) esophageal transit was investigated scintigraphically with a multiple swallow test protocol. 10/15 patients had a history of dysphagia. Patients were studied twice: under baseline conditions, and immediately after pharmacological stimulation with 10 mg of edrophonium chloride (EC), a short-acting acetylcholinesterase inhibitor. RESULTS: Under baseline conditions all patients showed an impaired esophageal function (emptying [%]= 58 % +/- 21; normal range > 85 %). In 14/15 individuals esophageal transit improved after administration of EC (emptying [%]= 75 % +/- 18; p < 0.01), reaching the normal range in 6 patients. One patient showed no effect attributable to EC. CONCLUSIONS: Esophageal transit is often compromised in MG. Functional abnormalities may be also present in patients without a history of dysphagia. Inhibition of cholinesterase positively affects striated muscles in the pharynx and upper esophagus, thus improving esophageal transit. Esophageal scintigraphy may be considered as a simple, non-invasive method for diagnosing impairment of esophageal function in MG and to monitor the changes under pharmacological stimulation. PMID- 12736743 TI - Investigation of a genetic variation of a variable number tandem repeat polymorphism of interleukin-6 gene in patients with multiple sclerosis. AB - Interleukin-6 (IL-6) plays an important role in the regulation of the inflammatory response in multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE). Previous reports indicated that the C allele of a variable number tandem repeat (vntr) polymorphism located in the 3'flanking region of the IL-6 gene ( IL-6) is associated with reduced activity of IL-6 in vivo. Since disease-modifying genes are likely to contribute to phenotypic differences in MS patients, we tested the hypothesis that the IL-6 C allele is associated with the clinical course of MS. The IL-6 C allele was equally distributed between 217 MS patients of German Caucasian origin and 111 age-mached healthy controls. Stratification of patients according to the course of disease revealed no significant difference of IL-6 C allele distribution between patients with primary progressive and those with either relapsing remitting or secondary progressive MS although IL-6 C allele was more frequent in patients with RR-MS. Since IL-6 C allele has been associated with a benign course in Sardinian MS patients, we further analysed an independent sample of 125 Sardinian MS patients revealing that IL-6 C allele was much more frequent than in German MS patients. Taken together, a disease-modifying effect of IL-6 C allele could not be demonstrated in MS patients of German Caucasian descent. PMID- 12736745 TI - Hearing improvement after tumor removal in a vestibular schwannoma patient with severe hearing loss. AB - We herein report on the hearing improvement obtained after tumor removal in a vestibular schwannoma patient with preoperative severe hearing loss and good otoacoustic emission. Hearing recovery after tumor removal is an uncommon occurrence. As a result, hearing preservation surgery is usually not performed for patients with severe hearing loss. However, patients with a good OAE seem to have a better chance to recover their hearing than those with a poor OAE, even if they have severe hearing loss. As a result, the cochlear function should be evaluated, especially regarding the OAE, before determining the indications for hearing preservation surgery. PMID- 12736746 TI - Two occult skull base malformations causing recurrent meningitis in a child: a case report. AB - Occult malformations of the skull base are rare anomalies, but can cause severe complications such as meningitis. Detailed skull base investigations for detecting cerebrospinal fluid fistulas or celes are often not initiated until after a history of recurrent meningitis. We present a child first seen at the age of 12 with recurrent episodes of bacterial meningitis since early childhood, requiring antibiotic prophylaxis for years. High-resolution computed tomography revealed a chronic sinusitis and a bony defect on the right olfactory groove, while magnetic resonance imaging and CT-cisternography indicated no cerebrospinal fluid fistula or cele at that time. Endonasal surgery for chronic sinusitis was performed with a confirmed bony defect on the right olfactory groove and an olfactory fibre without its sleeve-like dura prolongation running into an adjacent ethmoidal cell, necessitating that it be covered. In the absence of any antibiotics a new episode of meningitis occurred 5 years after surgery. CT cisternography and magnetic resonance imaging were repeated, now indicating a transclival bony defect with a meningocele in its proximal part, most probably presenting a canalis basilaris medianus. Endonasal surgery confirmed this bony defect after adenoidectomy, and closure was accomplished. No further meningitis has been observed for 2 years. Congenital skull base defects may be difficult to detect, but sufficient surgical closure after their precise delineation is mandatory to prevent infectious endocranial complications. The presence of more than one developmental skull base defect should be considered during careful radiological skull base evaluation, which has to include the clivus in order not to overlook rare basilar malformations. PMID- 12736744 TI - Molecular pathogenesis of head and neck squamous cell carcinoma. AB - Head and neck squamous cell carcinoma (HNSCC) represents 6% of all cancers. The overall 5-year survival rate for patients with this type of cancer is among the lowest of the major cancer types and has not improved dramatically during the last decade. The pathological staging, in particular the nodal stage, is the most important factor in HNSCC. The lack of progress in head and neck oncology emphasizes the importance of molecular genetic studies to define alterations that may correlate with tumor behavior. The molecular alterations observed in HNSCC are mainly due to oncogene activation and tumor suppressor gene inactivation, leading to deregulation of cell proliferation. These alterations include gene amplification and overexpression of oncogenes such as ras, myc, EGFR and cyclin D1, and mutations and deletions leading to p16 and TP53 tumor suppressor genes inactivation. This article reviews the molecular changes commonly observed in HNSCC. The biological function of these markers and the potential clinical application are discussed. Advances in the understanding of the molecular basis of HNSCC will help in the identification of new molecular markers that could be used for a more accurate diagnosis and assessment of prognosis and may open the way for novel approaches to treatment and prevention. PMID- 12736748 TI - Capillary changes in skeletal muscle of patients with Ullrich's disease with collagen VI deficiency. AB - We examined the capillaries in muscle biopsy specimens from two patients with Ullrich's disease with collagen VI deficiency by light and electron microscopy. Collagen VI plays an important role in platelet aggregation for binding von Willebrand factor. Using immunohistochemistry, collagen VI was shown to be absent on capillaries from patients with Ullrich's disease, while von Willebrand factor, collagen IV, and vascular endothelial growth factor were normally expressed. Electron microscopy revealed narrow lumens, large nuclei in endothelial cells, and fenestration of a capillary. The number of pinocytotic vesicles per unit endothelial cytoplasm was increased. The cytoplasm of endothelial cells was strongly stained with uranyl acetate and lead citrate. Replication of the capillary basement membrane was observed. On the other hand, easy bleeding and coagulation were not observed in the two patients. These findings suggested that the collagen VI deficiency might have caused the electron microscopic changes of capillaries, while the function of the capillaries is apparently retained. PMID- 12736747 TI - Autogenous fat injection for vocal fold atrophy. AB - Autogenous fat augmentation for glottic insufficiency has been used previously in patient treatment regimes. However, relatively little information is currently available regarding the effectiveness of fat injection for patients with vocal fold atrophy who have complete glottal closure (VFACGC). This paper compares, in retrospect, the efficiency of fat injection after surgery in patients with VFACGC (n=21). The perceptual acoustic and phonatory functions and videolaryngostroboscopic data were evaluated before and after fat augmentation was performed on 13 patients. Mean follow-up time was 9.5 months. Fifteen patients displayed excellent results, four experienced post-procedure failure, and two were unavailable for follow-up analysis. The majority of VFACGC patients (71%) also suffered from muscular tension dysphonia (MTD) preoperatively. The procedure also resolved the MTD in half (54%) of the patients in this study. Perceptual rating showed significant improvement in grade, roughness and breathiness (P<0.05). The videolaryngostroboscopic rating showed significant improvements in vocal fold edge linearity (P<0.01), vocal fold vibration amplitude and mucosal wave excursion (P<0.05). VFACGC is commonly misdiagnosed as MTD and is, therefore, unresponsive to speech therapy that is targeted to the latter. Fat injection is an effective autogenous implant and should be considered as an option in the treatment of patients with VFACGC. Although fat re-absorption was identified as a problem, repeating the procedure could be considered to minimize the effect of such. PMID- 12736749 TI - Extracorporeal shock-wave lithotripsy for treatment of ureteral calculi in paediatric patients. AB - Our objective was to determine the efficacy of extracorporeal shock wave lithotripsy (ESWL) in the treatment of paediatric ureteral calculous disease. We reviewed the records of 41 (23 boys, 18 girls) paediatric patients admitted to our clinic for the treatment of ureteral calculi between between 1989-2001. Patients' age varied between 4-16 years. The majority of the patients, 38 (92.7%) cases were treated initially with ESWL whereas 3 (7.3%) cases were subjected to ureterolithotomy. The mean age of the patients was found as 12.8+/-3.86 (4-16) years. Most calculi were located at either upper or lower ureter. The mean stone burden was 45.16+/-30.65 mm2 and the mean shock wave number per session and power as 2826.72+/-605.18 and 17.69+/-1.11 kV respectively. Minor complications included skin ecchymosis at the site of entry of shock waves in all cases and renal colic that responded to analgesics and emetics in 3 (7.9%) patients. The overall stone-free rate after ESWL was found to be 81.6%. Two (5.3%) cases have residual fragments that escaped to lower calices after lithotripsy for upper ureteral calculi and are still followed. There were 5 (13.2%) failures who were treated with ureterolitotomy for 1 upper and 1 lower ureteral calculi and with ureteroscopy for the rest. It appears that ESWL is still a good option for the initial treatment of most ureteral calculi in children as it is less invasive than ureteroscopy and has a high success rate as a first-line therapy. PMID- 12736750 TI - Foreign body aspiration in children: diagnosis and treatment. AB - A total of 235 children, aged between 7 months and 15 years had bronchoscopy on suspicion of foreign body aspiration. The histories of these patients were studied to examine the diagnostic value of symptoms, signs, and chest x-rays, and rate of negative bronchoscopy. The sensitivity of choking and coughing was high (82% and 80%), but the specificity was poor (37% and 34%). The sensitivity of a chest radiograph was 66%, the specificity was 51%. The sensitivity of asymmetric auscultation was 80% and specificity was 72%. The sensitivity and specificity of combination of symptoms, signs and abnormal chest radiograph was 61% and 83%, respectively. In 206 (87.7%) children a foreign body was identified and extracted. The remaining 29 patients (12.3%) had negative bronchoscopy. A wide variety of objects was recovered, the most common being seeds and peanuts. Foreign bodies were in the right and left main bronchus in 72 (35%), 50 (24.3%) cases, respectively, while in the remaining 84 cases, the foreign bodies were in other parts of the respiratory tree. In 204 (99%) patients with foreign body aspiration, the foreign bodies were removed successfully using a rigid bronchoscopy. Minor complications like subglottic edema and bronchospasm occurred in 4 children. In conclusion, rigid bronchoscopy is a safe procedure and the only tool that will give certainty about the correct diagnosis of foreign body aspiration in children. Asymmetric auscultation is more specific than history and chest radiograph. The combination of history, clinical signs and radiological signs are more specific than each one separately. PMID- 12736751 TI - Changes in cell type composition and enzymatic activities in the hepatopancreas of Marsupenaeus japonicus during the moulting cycle. AB - The goal of the present study was to evaluate the changes in the cell type composition and ATPase activities (total ATPase, ouabain-sensitive Na+/K(+) ATPase, furosemide-sensitive Na(+)-ATPase) that occur during the different stages of the moulting cycle in the hepatopancreas of the Marsupenaeus japonicus. The results clearly suggest that the number of resorptive and fibrillar cell types changes significantly during the different stages. An inverse correlation between resorptive and fibrillar cells is observed during moulting (both in normally fed and fasted animals). Fasting, but not the moulting cycle, affects the number of blister-like cells. In the resorptive cells the enzymatic activities (total ATPases and ouabain-sensitive Na+/K(+)-ATPase) also change during the moulting in a cyclical manner. All these results are in agreement with and confirm the different functions carried out by the two cell types within the hepatopancreas. PMID- 12736754 TI - Effect of contrast injection protocol with dose tailored to patient weight and fixed injection duration on aortic and hepatic enhancement at multidetector-row helical CT. AB - The aim of this study was to investigate the effect of a contrast material injection protocol with dose and injection rate of contrast material tailored to patient weight (dose tailored to patient weight and fixed injection duration). Hepatic helical CT was performed in 92 patients with chronic liver damage with a dose of 1.4 ml (518 mgI) at a rate of 0.056 ml/s per kilogram body weight of Iopamidol 370. Attenuation values of liver and aorta were measured for calculation of maximum aortic and hepatic enhancement, time to maximum hepatic enhancement, and end of hepatic arterial phase. Correlation coefficients between the injection rate and the four parameters were r=0.008, 0.057, 0.167, and 0.036, and there were no statistically significant correlations between the injection rates and the four parameters. In our injection protocol, uniform temporal scan window may be achieved and the injection rate can be reduced in lighter patients without reducing the degree of enhancement in the aorta and the liver. PMID- 12736755 TI - The role of the interventional radiologist in enteral alimentation. AB - The provision of enteral nutrition through the placement of gastrostomy/gastrojejunostomy tubes is a well-established procedure. Traditionally, these catheters have been placed either surgically or endoscopically; however, over the past two decades interventional radiologists have increasingly performed these procedures successfully. The perceived advantages of this route lie in the reported lower morbidity and mortality rates. In addition, percutaneous radiologically guided (PRG) catheters may be placed in certain subgroups of patients in whom it would be technically difficult or impossible by other routes, e.g., patients with head and neck or oesophageal tumours. The aim of this review is to describe the techniques of radiologically placed gastrostomy/gastrojejunostomy, discuss its indications and contraindications, describe any associated potential complications and compare PRG results with the more established techniques of open surgical and endoscopic placement. We also describe some recent procedural and catheter modifications. PMID- 12736756 TI - Ultrasonographic localization of occult pulmonary nodules during video-assisted thoracic surgery. AB - The aim of our study was to evaluate the role of ultrasonography in the localization of pulmonary nodules during video-assisted thoracic surgery (VATS). Ultrasonography was performed in 35 patients for the localization of pulmonary nodules during VATS. Indication for VATS was excisional biopsy of undetermined nodules in 22 patients, single or multiple metastasectomy in 12 patients and resection of primitive pulmonary cancer in 1 patient with reduced pulmonary reserve. A laparoscopic probe with flexible head and multi-frequency transducer (5-7.5 MHz) was used. Intraoperative ultrasonography localized 37 of 40 nodules preoperatively detected by CT and/or by positron emission tomography in 35 patients. Furthermore, ultrasonography localized two nodules not visualized at spiral CT. Eighteen nodules were not visible or palpable at thoracoscopic examination and were found by intraoperative sonography only. In 6 patients in whom thoracotomy was performed, manual palpation did not reveal more lesions than ultrasonography. In our experience, ultrasonography was very helpful when lesions were not visible or palpable during thoracoscopy, showing high sensitivity (92.5%) in finding pulmonary nodules. Since it is not possible to determine preoperatively whether a localization technique will be necessary during the operation or not, and ultrasonography is a non-invasive technique, we think that, at present, this technique can be considered as the first-instance localization technique during thoracoscopic resection of pulmonary nodules. PMID- 12736757 TI - The contribution of ultrasound for the differential diagnosis of congenital and infantile nephrotic syndrome. AB - The aim of this study was to determine whether high-resolution ultrasound is able to differentiate between the various diseases associated with nephrotic syndrome (NS). We reviewed the US features of 15 patients less than 1 year presenting a NS whose exact type was defined by pathology nephrotic syndrome of Finnish type (NSFT, n=2); focal and segmental hyalinosis (FSH, n=3); minimal-change glomerular disease (MCGD, n=2); neonatal glomerulonephritis (n=1), and diffuse mesangial sclerosis (DMS, n=7). The US features studied included the size of the kidneys, cortical echogenicity, cortico-medullary differentiation (CMD), and borders. The images were reviewed on hard copies by two observers unaware of the final diagnosis. In each case a diagnosis was proposed based on the reading of the US features. Six patients with DMS displayed a peculiar US pattern: mild increase of renal size; and inhomogeneous (patchwork-like) parenchymal hyperechogenicity that included areas of the cortex and medulla. The NSFT and neonatal glomerulonephritis displayed some of the same US features: increased kidney size (+2 SD) and had homogeneous cortical hyperechogenity with persistent cortico medullar differentiation. The kidneys in the 3 patients with SFH were sonographically normal (n=1) or displayed a mild cortical hyperechogenicity (n=2). Inhomogeneous parenchymal hyperechogenicity involving only segments of the cortex and medulla seems to be a specific US pattern for DMS. Ultrasound is less specific for the other types of CNS. PMID- 12736759 TI - All- trans-retinoic acid increases cytotoxicity of 1-beta-D arabinofuranosylcytosine in NB4 cells. AB - PURPOSE: Clinically, the benefits of combining all- trans-retinoic acid (ATRA) with chemotherapy have been well documented in the treatment of acute promyelocytic leukemia (APL). Changes in nucleoside transporter expression and activity have been shown to occur in NB4 cells in vitro following treatment with ATRA. In this study we investigated whether ATRA treatment increases sensitivity to ara-C in NB4 cells. Specifically, we examined the role of ATRA-associated changes in nucleoside transporter expression and activity in eliciting ara-C cytotoxicity. METHODS: Cellular uptake of [(3)H]-ara-C and nucleoside transporter abundance were determined in untreated cells and cells treated with 1 microM ATRA for 12-72 h using an inhibitor and oil stop procedure, and an equilibrium [(3)H] NBMPR binding assay, respectively. Cytotoxicity of ara-C and the apoptotic response prior to and following ATRA treatment were determined using the MTT viability assay and the TUNEL assay, respectively. RESULTS: ATRA treatment increased ara-C cytotoxicity and potency, ara-C transport, and augmented ara-C induced apoptosis. The combination effect was supraadditive under some conditions and sequence-dependent whereby the maximum effect was seen when the addition of ATRA preceded the addition of ara-C, and when ara-C administration closely followed ATRA administration. CONCLUSIONS: The ATRA-induced increase in cytotoxicity of ara-C was, in part, the result of an increase in the functional expression of nucleoside transporters, and a role for bcl-2 was also indicated. Our results would suggest that timing of ara-C therapy should be tied to maximal es transporter expression, which is likely to be 24 h after ATRA treatment begins. It remains to be seen whether the response in the clinic can be further enhanced in APL by taking advantage of ara-C transporter regulation by ATRA. PMID- 12736760 TI - In vivo antitumor activity of S16020, a topoisomerase II inhibitor, and doxorubicin against human brain tumor xenografts. AB - New active drugs are needed for the treatment of primary brain tumors in both children and adults. S16020 is a cytotoxic olivacine derivative that inhibits topoisomerase II. The aim of the study was to determine its antitumor activity in athymic mice bearing subcutaneous medulloblastoma (IGRM33, 34, 57) and glioblastoma (IGRG88, 93, 121) xenografts treated at an advanced stage of tumor growth in comparison with that of doxorubicin. Animals were randomly assigned to receive i.v. S16020 or doxorubicin weekly for three consecutive weeks. The optimal dose was 80 mg/kg per week. S16020 demonstrated a significant antitumor activity in two out of three medulloblastoma xenografts. IGRM57 xenografts were highly sensitive with 100% tumor regressions and a tumor growth delay (TGD) of 102 days, while one of eight IGRM34 xenografts showed a partial regression with a TGD of 16 days. Doxorubicin was significantly more active than S16020 in these two models. IGRM33, a model established from a tumor in relapse after chemotherapy and radiotherapy, was refractory to both drugs. S16020 demonstrated a significant antitumor activity in the three glioblastoma xenografts evaluated. The wild-type p53 IGRG93 xenograft was highly sensitive with 100% tumor regressions and a TGD of 54 days. IGRG121 (wt p53) and IGRG88 (mutant p53) were moderately sensitive with TGDs of 33 and 23 days, respectively. Doxorubicin showed greater activity in two of these models. All six xenografts exhibited low expression of mdr1 as quantitated by RT-PCR, and no correlation was found with the activity of either drug. Conversely, a low activity of the two drugs was significantly associated with a high expression of MRP1 in medulloblastomas. Finally, no relationship was observed between drug sensitivity to either drug and expression of their target, topoisomerase IIalpha. In conclusion, S16020 and doxorubicin showed significant antitumor activity in brain tumor xenografts treated at an advanced stage of tumor growth. Their activity was related to MRP1 expression in medulloblastomas. PMID- 12736758 TI - Evolution, biochemistry and genetics of protein kinase C in fungi. AB - From yeast to humans, protein kinase C is an enzyme of central importance in signal transduction processes. In baker's yeast, Saccharomyces cerevisiae, a single gene encoding Pkc1p has been identified. Mutant analyses revealed that Pkc1p function is essential to ensure cellular integrity through regulation of a specific MAP kinase cascade. Due to the involvement of different defective mammalian isozymes in various diseases and the model character of simple eukaryotes, increasing attention has been paid to the structure and regulation of the enzymes of fungal origin. This review summarizes the knowledge gathered so far. PMID- 12736761 TI - In vitro activity of dimethylarsinic acid against human leukemia and multiple myeloma cell lines. AB - PURPOSE: Arsenic trioxide (As(2)O(3)), an inorganic arsenic compound, has recently been approved for the treatment of relapsed or refractory acute promyelocytic leukemia. However, systemic toxicity associated with As(2)O(3) treatment remains a problem. Inorganic arsenic is detoxified in vivo by methylation reactions into organic arsenic compounds that are less toxic. METHODS AND RESULTS: We investigated the antiproliferative and cytotoxic activity of dimethylarsinic acid (DMAA), an organic arsenic derivative and major metabolic by product of As(2)O(3), against a panel of eight leukemia and multiple myeloma cell lines. As(2)O(3) was tested in comparison. In clonogenic assay, the average concentration of DMAA that suppressed cell colony growth by 50% was 0.5-1 m M, while for As(2)O(3) it was on average 1-2 microM. At those concentrations DMAA and As(2)O(3) had significantly less effect on colony growth of normal progenitor cells. Cytotoxic doses of DMAA and As(2)O(3) in 3-day trypan blue dye exclusion assay experiments were similar to doses effective in clonogenic assay. Assessment of apoptosis by annexin V assay revealed a high rate of apoptosis in all cell lines treated with DMAA and As(2)O(3), but significantly less effect on normal progenitor cells. DMAA, unlike As(2)O(3), had no effect on the maturation of leukemic cells. CONCLUSIONS: DMAA exerts differential antiproliferative and cytotoxic activity against leukemia and multiple myeloma cells, with no significant effect on normal progenitor cells. However, concentrations of DMAA needed to achieve such efficacy are up to 1000 times those of As(2)O(3). Evaluation of novel organic arsenic that would combine the high efficacy of As(2)O(3) and the low toxicity of DMAA is warranted. PMID- 12736763 TI - Clinical and pharmacologic study of tributyrin: an oral butyrate prodrug. AB - PURPOSE: Butyrate is a small polar compound able to produce terminal differentiation and apoptosis in a variety of in vitro models at levels above 50 100 microM. Previously our group demonstrated that daily oral administration of the prodrug, tributyrin, is able to briefly achieve levels >100 microM. Given in vitro data that differentiating activity requires continuous butyrate exposure, the short t1/2 of the drug and a desire to mimic the effects of an intravenous infusion, we evaluated a three times daily schedule. PATIENTS AND METHODS: Enrolled in this study were 20 patients with advanced solid tumors for whom no other therapy was available, had life expectancy greater than 12 weeks, and normal organ function. They were treated with tributyrin at doses from 150 to 200 mg/kg three times daily. Blood was sampled for pharmacokinetic analysis prior to dosing and at 15 and 30 min and 1, 1.5, 2, 2.5, 3, 3.5 and 4 h thereafter. RESULTS: The patients entered comprised 15 men and 5 women with a median age of 61 years (range 30-74 years). Prior therapy regimens included: chemotherapy (median two prior regimens, range none to five), radiation therapy (one), no prior therapy (one). There was no dose-limiting toxicity. Escalation was halted at the 200 mg/kg three times daily level due to the number of capsules required. A median butyrate concentration of 52 microM was obtained but there was considerable interpatient variability. No objective responses were seen. There were four patients with prolonged disease stabilization ranging from 3 to 23 months; median progression-free survival was 55 days. Two patients with chemotherapy-refractory non-small-cell lung cancer had survived for >1 year at the time of this report without evidence of progression. CONCLUSION: Tributyrin is well tolerated and levels associated with in vitro activity are achieved with three times daily dosing. PMID- 12736762 TI - A phase II and pharmacokinetic study of pegylated liposomal doxorubicin in patients with advanced hepatocellular carcinoma. AB - PURPOSE: Chemotherapy of advanced hepatocellular carcinoma (HCC) is frequently limited by unacceptable toxicity. Long-circulating polyethylene glycol-coated (PEGylated) liposomal doxorubicin (PLD) has low systemic toxicity. Its safety and efficacy in patients with advanced HCC and the relationship between hepatic function and pharmacokinetics were investigated in this phase II study. METHODS: Patients were given 30 mg/m(2) PLD every 3 weeks and the dose was escalated to 45 mg/m(2) from the third course if the toxicity was deemed tolerable. The plasma level of doxorubicin was determined with fluorometry. RESULTS: A total of 40 patients were recruited into this phase II study. The toxicities were usually mild but unexpectedly, three cirrhotic patients died of infection without neutropenia. Four had a partial response (response rate 10%, confidence interval 0-20%). The median duration of response was 5.6 months. The median time to tumor progression and the median survival of all patients was only 2 and 3 months, respectively. Patients with advanced HCC had lower initial serum concentration, larger volume of distribution and more rapid clearance than patients with other malignancies and normal liver function. However, the pharmacokinetic parameters correlated with neither toxicity nor response. CONCLUSIONS: The disposition of PLD in patients with liver dysfunction was not hampered, but it did not exhibit higher activity compared with free drug, and the risk of infection must be watched closely especially in patients with liver cirrhosis. PMID- 12736765 TI - The stability of lidocaine and epinephrine solutions exposed to electric current and comparative administration rates of the two drugs into pig bladder wall. AB - Intravesical electromotive administration of local anesthetics is clinically successful but electrochemistry, cost and effectiveness limit the choice of drugs to diluted lidocaine HCl 4% mixed with epinephrine. These studies address the stability of lidocaine and epinephrine both over time and when exposed to electric current, i.e. transport rates with passive diffusion and electromotive administration. The drug mixture used was 50 ml lidocaine 4%, 50 ml H2O and 1 ml epinephrine 1/1000. For stability, the solution was placed either in bowls for 7 days or in a two chamber cell with the donor compartment (drugs) separated from the receptor compartment (NaCl solution) by a viable pig bladder wall. This was subjected to 30 mA for 45 min. Stability was measured with mass spectrometry. The cell was also used to determine transport rates with passive diffusion and currents of 20 mA and 30 mA, over 20, 30 and 45 min. Drug measurements in both compartments and bladder were made with HPLC. Lidocaine remained stable throughout the 7 days, epinephrine on day 1 only and both drugs were stable with 30 mA for 45 min. Comparing 20 mA and 30 mA with passive diffusion, there were significant differences in 6/6 donor compartment lidocaine levels, 4/6 receptor compartment levels and 6/6 bladder tissue levels and also in 6/6 epinephrine donor levels and 6/6 tissue levels. The combination lidocaine and epinephrine remains stable for 1 day and when exposed to 30 mA for 45 min. Electric current accelerates the transport of lidocaine and epinephrine. PMID- 12736766 TI - The role of cholinesterases in rat urinary bladder contractility. AB - This study examines the effects of inhibition of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) on acetylcholine (ACh)-induced contraction in rat urinary bladder smooth muscle. Neostigmine, a non-selective ChE inhibitor, caused concentration-dependent contractions in rat urinary bladder strips, whereas tetraisopropylpyrophosphoramide (iso-OMPA; a BuChE inhibitor) failed to affect the resting tone of the preparations. Neostigmine (1 microM) markedly augmented the contractile responses to ACh. Although iso-OMPA (10 microM) also potentiated ACh-induced contraction, the effect was less than that evoked by neostigmine. The activities of AChE in rat urinary bladder strips were significantly (P<0.05) higher than those of BuChE. These results indicated that AChE, rather than BuChE, plays an important role in controlling ACh-induced contractions of rat urinary bladder. PMID- 12736767 TI - MR diffusion imaging and MR spectroscopy of maple syrup urine disease during acute metabolic decompensation. AB - Maple syrup urine disease (MSUD) is an inborn error of amino acid metabolism, which affects the brain tissue resulting in impairment or death if untreated. Imaging studies have shown reversible brain edema during acute metabolic decompensation. The purpose of this paper is to describe the diffusion-weighted imaging (DWI) and spectroscopy findings during metabolic decompensation and to assess the value of these findings in the prediction of patient outcome. Six patients with the diagnosis of MSUD underwent conventional MR imaging with DWI during acute presentation with metabolic decompensation. Spectroscopy with long TE was performed in four of the six patients. Follow-up examinations were performed after clinical and metabolic recovery. DWI demonstrated marked restriction of proton diffusion compatible with cytotoxic or intramyelinic sheath edema in the brainstem, basal ganglia, thalami, cerebellar and periventricular white matter and the cerebral cortex. This was accompanied by the presence of an abnormal branched-chain amino acids (BCAA) and branched-chain alpha-keto acids (BCKA) peak at 0.9 ppm as well as elevated lactate on proton spectroscopy in all four patients. The changes in all six patients were reversed with treatment without evidence of volume loss or persistent tissue damage. The presence of cytotoxic or intramyelinic edema as evidenced by restricted water diffusion on DWI, with the presence of lactate on spectroscopy, could imply imminent cell death. However, in the context of metabolic decompensation in MSUD, it appears that changes in cell osmolarity and metabolism can reverse completely after metabolic correction. PMID- 12736769 TI - Recent advances in the applications of forensic science to fire debris analysis. AB - The forensic discipline of ignitable liquid and fire debris analysis is rapidly changing. Refinements in existing methods as well as development of new techniques are changing the routine methods of analysis. Optimization of existing extraction techniques and research into novel methods of extracting debris have improved the recovery of ignitable liquids from debris samples. The application of highly specialized instrumentation to problems of sensitivity and matrix interference has resulted in new ways of performing chemical analyses, allowing for improved limits of detection. Preliminary research in novel approaches to ignitable liquid comparisons is being evaluated, with the hopes of providing more detailed information to the field investigators. Research into a variety of areas related to fire debris analysis is ongoing, and will continue to improve the quality of ignitable liquid residue analysis. PMID- 12736768 TI - MRI of intracerebral hematoma: value of vasogenic edema ratio for predicting the cause. AB - We evaluated a method for quantifying vasogenic edema (VE) on MRI to identify brain hematomas caused by neoplasms. We performed a blinded review of 68 acute and subacute hematomas caused by neoplasms (22), hypertension or presumed amyloid angiopathy (27), or vascular malformations (19). The extent of vasogenic edema was quantified on an axial T2-weighted image using the VE ratio: the maximum width of high signal surrounding a hematoma and the mean diameter of the hematoma. Hematomas caused by neoplasm were associated with more vasogenic edema (mean VE ratio 104%+/-15%; mean VE width 2.4+/-0.7 cm) than non-neoplastic hematomas (mean VE ratio 37%+/-5%; P =0.001). When the width of VE was equal to or more half the diameter the hematoma (VE ratio 50%), the positive predictive value for tumor was 66%; when it was equal to or more than the diameter, the positive predictive value was 71%. All six hematomas with VE ratios > or = 150% were caused by neoplasm. PMID- 12736771 TI - Novobiocin biosynthesis: inactivation of the putative regulatory gene novE and heterologous expression of genes involved in aminocoumarin ring formation. AB - The left ends of the biosynthetic gene clusters of novobiocin ( nov), clorobiocin ( clo) and coumermycin A(1) ( cou) from Streptomyces spheroides (syn. S. caeruleus) NCIMB 11891, S. roseochromogenes var. oscitans DS 12.976 and S. rishiriensis DSM 40489 were cloned and sequenced. Sequence comparison suggested that novE, cloE and couE, respectively, represent the borders of these three clusters. Inactivation of novE proved that novE does not have an essential catalytic role in novobiocin biosynthesis, but is likely to have a regulatory function. The gene products of novF and cloF show sequence similarity to prephenate dehydrogenase and may produce 4-hydroxyphenylpyruvate (4HPP) as a precursor of the substituted benzoate moiety of novobiocin and clorobiocin. Coumermycin A(1) does not contain this benzoate moiety, and correspondingly the coumermycin cluster was found not to contain a functional novF homologue. The coumermycin biosynthetic gene cluster apparently evolved from an ancestral cluster similar to those of novobiocin and clorobiocin, and parts of the ancestral novF homologue have been deleted in this process. No homologue to novC was identified in the gene clusters of clorobiocin and coumermycin, questioning the postulated involvement of novC in aminocoumarin biosynthesis. Heterologous expression of novDEFGHIJK in Streptomyces lividans resulted in the formation of 2,4-dihydroxy-alpha-oxy-phenylacetic acid, suggesting that at least one of the proteins encoded by these genes may participate in a hydroxylation reaction. PMID- 12736770 TI - BIMU 1 and RS 67333, two 5-HT4 receptor agonists, modulate spontaneous alternation deficits induced by scopolamine in the mouse. AB - The present study was conducted to determine the effects of two potent 5-HT4 receptor agonists, BIMU 1 (1 (3-ethyl-2,3-dihydro-N-[endo-8-methyl-8-azabicyclo (3.2.1)-oct-3-yl]-2-oxo-1H) benzimidazole-1-carboxamide hydrochloride; 1, 3, 10 mg/kg, i.p.) and RS 67333 (1-(4-amino-5-chloro-2-methoxyphenyl)-3-(1-n-butyl-4 piperidinyl)-1-propanone; 0.25, 0.5, 1 mg/kg, i.p.) on the learning impairment induced by the muscarinic acetylcholine receptor antagonist, scopolamine (1 mg/kg) in mice. Working memory was examined by observing spontaneous alternation behavior in the Y-maze test. Both BIMU 1 (10 mg/kg) and RS 67333 (1 mg/kg) prevented the scopolamine-induced alternation deficits, whereas no effect could be evidenced on locomotor or emotional indices. The reversal actions of BIMU 1 and RS 67333 on this cognitive dysfunction were abolished by the selective 5-HT4 receptor antagonist GR 125487 (1-[2-[(methyl sulfonyl)-amino]-ethyl]-4 piperidinyl-methyl-5-fluoro-2-methoxy-1H-indole-3-carboxylate; 10 mg/kg, i.p.). When given alone at the same doses, none of the three serotonergic agents had any measurable effect. These results demonstrate the ability of 5-HT4 receptor agonists to reverse spontaneous working memory deficits and further confirm the therapeutic potential of such ligands in the treatment of cognitive alterations that associate short-term working memory disorders and cholinergic hypofunction. PMID- 12736772 TI - HMG CoA reductase inhibitors and the skeleton: a comprehensive review. AB - Recent studies suggest that the mevalonate pathway plays an important role in skeletal metabolism. HMG CoA reductase inhibitors ("statins"), which inhibit a key enzyme in the mevalonate pathway, are widely used for the treatment of hyperlipidemia. In vitro and animal studies demonstrate that statins stimulate the production of BMP-2, a potent regulator of osteoblast differentiation and activity, suggesting that statins may have an anabolic effect on bone. Statin use in most, but not all observational studies is associated with a reduced risk of fracture, particularly hip fracture, even after adjustment for the confounding effects of age, weight and other medication use. This beneficial effect has not been observed in clinical trials designed to assess cardiovascular endpoints. The effects of statins on bone mass and bone turnover are controversial, but increased bone mass and reduced bone turnover have been observed in controlled studies. Further studies of the skeletal effects of statins are needed, particularly their effects on surrogate markers such as bone mass, bone turnover, and microarchitecture, to determine the optimal formulation, dosing and route of administration. Clinical trials with fracture endpoints are needed before statins can be recommended as therapeutic agents for osteoporosis. PMID- 12736773 TI - Failure strength of bioabsorbable interference screws: effects of in vivo degradation for 12 weeks. AB - We investigated the effect of in vivo degradation for 6-12 weeks on the fixation strength of polylactide bioabsorbable interference screws. Ten bioabsorbable interference screws were used to fix the patellar tendon autograft in ten live sheep knees, which were equally divided into two groups and killed in the 6th or 12th week. The control group consisted of four cadaveric knees. Following the killing of the animals the screws were retrieved and reused to fix patellar tendon grafts in cadaveric sheep knees. Tendon pull-out tests were performed for the partially degraded screws, for the control group, and for the reused screws of the control group. Macroscopic and microscopic examinations of the 6- and 12 week specimens were performed. Tendons pulled-out with an average force of 357+/ 30 N in the cadaveric control group on the first use and with 465+/-118 N on the second use. The partially degraded screws failed with a mean load of 399+/-119 N in the 6-week group, and 12-week screws at 447+/-72 N. No macroscopic sign of degradation was observed on the retrieved screws. Histological examination of the 6 week specimens showed necrotic changes in the tendon around screw contact areas. Healing with granulation tissue was present in the same area in the 12th week. Foreign body reaction or an excessive inflammatory reaction was not observed. In vivo degradation of poly- l-lactide interference screws for 12 weeks thus causes neither a loss in the fixation strength of the screws nor an obvious inflammatory reaction. PMID- 12736775 TI - Effect of vasopressin on sublingual microcirculation in a patient with distributive shock. AB - OBJECTIVE: To assess the sublingual microcirculation in a patient during vasopressin administration for a distributive shock after cardiopulmonary bypass. DESIGN AND SETTING: Case-report in the Department of Intensive Care of a university hospital. PATIENT: A 53 year-old man developed severe distributive shock after cardiac transplant, requiring massive doses of vasopressor agents. METHODS: Vasopressin administered twice at a dose of 0.02 U/min increased mean blood pressure and allowed partial weaning of other vasopressor drugs. Microcirculatory alterations were assessed by orthogonal polarization spectral technique: 50% and 60% of capillaries were perfused at baseline, and these proportions did not worsen when vasopressin was administered. CONCLUSIONS: Despite its strong vasopressor effects vasopressin infusion did not worsen microcirculatory alterations in this patient with distributive shock following cardiac surgery. PMID- 12736776 TI - Pulmonary capillary pressure. PMID- 12736778 TI - Molecular dissection of a dormancy QTL region near the chromosome 7 (5H) L telomere in barley. AB - Moderate seed dormancy is desirable in barley (Hordeum vulgare L.). It is difficult for breeders to manipulate seed dormancy in practical breeding programs because of complex inheritance and large environmental effects. Quantitative trait locus (QTL) mapping opens a way for breeders to manipulate quantitative trait genes. A seed dormancy QTL, SD2, was mapped previously in an 8-cM interval near the chromosome 7 (5H) L telomere from a cross of 'Steptoe' (dormant)/'Morex' (non-dormant) by the North American Barley Genome Project using an interval mapping method and a relatively low-resolution genetic map. SD2 has a moderate dormancy effect, which makes it a promising candidate gene for moderate seed dormancy in barley cultivar development. The fine mapping of SD2 is required for efficient manipulation of SD2 in breeding and would facilitate the study of dormancy in barley. Ten different Morex isolines were generated, including regenerated Morex, of which nine lines had duplicates. The isolines together with Steptoe and Morex were grown in growth room and field environments for 2 years (2000 and 2001). In the growth room, relatively low growing temperatures (25 degrees C day/15 degrees C night) were employed to promote seed dormancy development. Seed germination percentage, determined at different post-harvest after-ripening periods, was used to measure seed dormancy. Fine mapping using the substitution mapping method based on differences among isolines resolved the SD2 QTL into an 0.8-cM interval between molecular markers MWG851D and MWG851B near the chromosome 7 (5H) L telomere. Relatively low temperatures (< or =25 degrees C) during seed development promoted the expression of the SD2 dormancy QTL. The chromosome region above the MWG851D-MWG851B interval might play a role in reducing barley seed dormancy during after-ripening. PMID- 12736777 TI - Mapping quantitative trait loci for yield, yield components and morphological traits in an advanced backcross population between Oryza rufipogon and the Oryza sativa cultivar Jefferson. AB - An advanced backcross population between an accession of Oryza rufipogon (IRGC 105491) and the U.S. cultivar Jefferson (Oryza sativa ssp. japonica) was developed to identify quantitative trait loci (QTLs) for yield, yield components and morphological traits. The genetic linkage map generated for this population consisted of 153 SSR and RFLP markers with an average interval size of 10.3 cM. Thirteen traits were examined, nine of which were measured in multiple environments. Seventy-six QTLs above an experiment-wise significance threshold of P<0.01 (corresponding to an interval mapping LOD>3.6 or a composite interval mapping LOD>3.9) were identified. For the traits measured in multiple environments, 47% of the QTLs were detected in at least two environments. The O. rufipogon allele was favorable for 53% of the yield and yield component QTLs, including loci for yield, grains per panicle, panicle length, and grain weight. Morphological traits related to the domestication process and/or weedy characteristics, including plant height, shattering, tiller type and awns, were found clustered on chromosomes 1 and 4. Comparisons to previous studies involving wild x cultivated crosses revealed O. rufipogon alleles with stable effects in multiple genetic backgrounds and environments, several of which have not been detected in studies between Oryza sativa cultivars, indicating potentially novel alleles from O. rufipogon. Some O. rufipogon-derived QTLs, however, were in similar regions as previously reported QTLs from Oryza sativa cultivars, providing evidence for conservation of these QTLs across the Oryza genus. In addition, several QTLs for grain weight, plant height, and flowering time were localized to putative homeologous regions in maize where QTLs for these traits have been previously reported, supporting the hypothesis of functional conservation of QTLs across the grasses. PMID- 12736780 TI - Sugar-induced adventitious roots in Arabidopsis seedlings. AB - The effects of sugars on root growth and on development of adventitious roots were analyzed in Arabidopsis thaliana. Seeds were sown on agar plates containing 0.0-5.0% sugars and placed vertically in darkness (DD) or under long day (LD, 16 h:8 h) conditions, so that the seedlings were constantly attached to the agar medium. In the sucrose-supplemented medium, seedlings showed sustained growth in both DD and LD. However, only dark-grown seedlings developed adventitious roots from the elongated hypocotyl. The adventitious roots began to develop 5 days after imbibition and increased in number until day 11. They could, however, be initiated at any position along the hypocotyl, near the cotyledon or the primary root. They were initiated in the pericycle in the same manner as ordinary lateral roots. Sucrose, glucose and fructose greatly stimulated the induction of adventitious roots, but mannose or sorbitol did not. Sucrose at concentrations of 0.5-2.0% was most effective in inducing adventitious roots, although 5.0% sucrose suppressed induction. Direct contact of the hypocotyl with the sugar-supplemented agar medium was indispensable for the induction of adventitious roots. PMID- 12736779 TI - A novel gene, RSD-3/HSD-3.1, encodes a meiotic-related protein expressed in rat and human testis. AB - The expression of stage-specific genes during spermatogenesis was determined by isolating two segments of rat seminiferous tubule at different stages of the germinal epithelium cycle delineated by transillumination-delineated microdissection, combined with differential display polymerase chain reaction to identify the differential transcripts formed. A total of 22 cDNAs were identified and accepted by GenBank as new expressed sequence tags. One of the expressed sequence tags was radiolabeled and used as a probe to screen a rat testis cDNA library. A novel full-length cDNA composed of 2228 bp, designated as RSD-3 (rat sperm DNA no.3, GenBank accession no. AF094609) was isolated and characterized. The reading frame encodes a polypeptide consisting of 526 amino acid residues, containing a number of DNA binding motifs and phosphorylation sites for PKC, CK II, and p34cdc2. Northern blot of mRNA prepared from various tissues of adult rats showed that RSD-3 is expressed only in the testis. The initial expression of the RSD-3 gene was detected in the testis on the 30th postnatal day and attained adult level on the 60th postnatal day. Immunolocalization of RSD-3 in germ cells of rat testis showed that its expression is restricted to primary spermatocytes, undergoing meiosis division I. A human testis homologue of RSD-3 cDNA, designated as HSD-3.1 (GenBank accession no. AF144487) was isolated by screening the Human Testis Rapid-Screen arrayed cDNA library panels by RT-PCR. The exon-intron boundaries of HSD-3.1 gene were determined by aligning the cDNA sequence with the corresponding genome sequence. The cDNA consisted of 12 exons that span approximately 52.8 kb of the genome sequence and was mapped to chromosome 14q31.3. PMID- 12736781 TI - Apomixis in the interspecific triploid hybrid fern Cornopteris christenseniana (Woodsiaceae). AB - Cornopteris christenseniana is a "sterile" interspecific triploid hybrid of diploid C. crenulatoserrulata and tetraploid C. decurrenti-alata. Morphological and cytological studies show that, of 41 young plants of Cornopteris that have been propagated naturally in the Fern Garden of the Botanical Gardens, University of Tokyo, 30 plants are the sterile C. christenseniana, 10 are fertile C. decurrenti-alata and 1, fertile C. crenulatoserrulata. This proportion supports the view that the young plants of C. christenseniana are derived from spores of reproductively mature plants of the species cultivated. Cytogenetic observations and culture experiments show that C. christenseniana produces normal spores in various proportions in some sporogenetic pathways that are aberrant from the ordinary process in sexual and apomictic ferns. Under culture conditions, normal spores germinate in rough proportion to the frequency of normal spores, and sporophytes are apogamously produced in rough proportion to the frequency of spore germination. As a whole, the rates of spore germination and apogamous sporophyte development vary according to the specific plant. Taken together, these observations suggest that C. christenseniana is an incipient apomict. PMID- 12736782 TI - Distyly and pollen dimorphism in Damnacanthus (Rubiaceae). AB - We document distyly in Damnacanthus (Rubiaceae) and investigate pollen morphology in the genus. Within nine taxa in Japan (four species, four varieties and one form), five show distyly. Non-distylous taxa of Damnacanthus always have a long style and short stamens. We examined the pollen morphology of all taxa of Damnacanthus in Japan. The pollen grains of Damnacanthus are 3- to 5 orthocolporate or sometimes loxocolporate. Polymorphic pollen is sometimes observed even within a single flower. Pollen dimorphism is associated with distyly; pollen grains from short-styled (thrum) flowers are significantly larger than those from long-styled (pin) flowers, and the muri are smooth in pin flowers whereas the pollen grains of thrum flowers have minute granules on the top and/or on the sides. In non-distylous taxa, in which the style is longer than stamens, the muri are always provided with minute granules. Interestingly, the pollen grains with granules in the non-distylous taxa contrast with the pollen of the distylous taxa, because granulate pollen grains are found only in thrum flowers. Although Damnacanthus is a stenopalynous genus, the number of colpi is useful for delimiting taxa. Based on palynological characters, we conclude that Damnacanthus should be revised. PMID- 12736783 TI - Phylogenetic relationships among genera of Massonieae (Hyacinthaceae) inferred from plastid DNA and seed morphology. AB - The tribe Massonieae Baker (Hyacinthaceae-Hyacinthoideae) presently consists of about 19 genera and 230 species distributed from Africa (south of the Sahara) to Madagascar and India. Based on atpB and trnL-F DNA sequences the tribe is monophyletic only when the genus Pseudoprospero is excluded from Massonieae. In most trnL-F trees, this genus occupies a basal position within subfamily Hyacinthoideae and is sister to the rest of the subfamily. Molecular data suggest that the remaining genera of Massonieae do not share common ancestry with the Eurasian/North-African tribe Hyacintheae Dumort. ( Scilla, Hyacinthus and allies), and thus a narrow concept of the essentially Eurasian genus Scilla is supported. Members of well-supported clades in Massonieae usually show similarities in seed characteristics as determined by scanning electron microscopy. Phylogenetic position and seed morphology indicate that Massonia angustifolia and M. zeyheri do not belong to the genus Massonia but fall into a clade together with Daubenya, Androsiphon and Amphisiphon. The genus Whiteheadia appears paraphyletic in the 50% majority rule trnL-F tree and occupies a basal position next to Massonia. However, in the strict consensus tree neither monophyly nor polyphyly can be excluded for this genus. Seed appendages are documented for members of the genera Ledebouria and Lachenalia. Within the genera of Massonieae there is a tendency towards bending of the seed axis. This phenomenon is most obvious within the genus Lachenalia. Delimitation of genera based on seed morphology largely agrees with the results of molecular studies. Correlation between number, size and color of seeds, geographical distribution and phylogenetic position of the genera are discussed. PMID- 12736784 TI - Purification and properties of betaine aldehyde dehydrogenase from Avena sativa. AB - Betaine aldehyde dehydrogenase (BADH; EC 1.2.1.8) is the enzyme that catalyzes the second step in the synthesis of the osmoprotectant, glycine betaine. NAD dependent BADH was purified from Avena sativa shoots by DEAE Sephacel, hydroxyapatite, 5'-AMP Sepharose 4B, Mono Q and TSK-GEL column chromatographies to homogeneity by the criterion of native PAGE, and the properties of BADH were compared with those of aminoaldehyde dehydrogenase purified to homogeneity from A. sativa. The molecular mass estimated by both gel filtration using TSK-GEL column and Sephacryl S-200 was 120 and 115, kDa, respectively. The enzyme is a homodimer with a subunit molecular mass of 61 kDa as shown by SDS-PAGE. The pI value of the enzyme was found to be 6.3. The purified enzyme catalyzed not only the oxidation of betaine aldehyde (BAL), but also that of aminoaldehydes, 3 aminopropionaldehyde (APAL), 4-aminobutyraldehyde (ABAL), and 4 guanidinobutyraldehyde (GBAL). The K(m) values for BAL, APAL, ABAL and GBAL were 5x10(-6), 5.4x10(-7), 2.4x10(-5) and 5x10(-5) M, respectively. APAL showed substrate inhibition at a concentration of 0.1 mM. A fragment of BADH cleaved by V8 protease shared homology with other plant BADHs. PMID- 12736785 TI - Megagametophyte abnormalities of near-isogenic female partial-sterile soybean mutants ( Glycine max; Leguminosae). AB - Megagametogenesis of soybean, Glycine max (L.) Merr., cultivars Clark and Clark k2, and F1 hybrid of Clark (female parent) crossed with Clark k2 (male parent) were studied using stereo light microscopy and confocal scanning laser microscopy. Reproductive development in Clark and Clark k2 plants was compared to F1 hybrid plants. In mature pods, 6.4% of the ovules of Clark, 8.1% of the ovules of Clark k2, and 41.4% of the ovules of F1 hybrid plants were aborted. This female partial sterility was due to incomplete megagametophyte development: undeveloped polar nuclei-or developed but not in a position for fertilization; increased megagametophyte wall thickness; abnormal shape and/or premature degeneration of synergids and intact synergids throughout the life of the ovule; egg cell not well-developed or absent; and megagametophyte remaining uninucleate. Each of these abnormalities contributed to either lack of double fertilization or early megagametophyte abortion. PMID- 12736786 TI - SLEEPLESS, a gene conferring nyctinastic movement in legume. AB - A genetic approach was attempted to identify the gene responsible for nyctinastic movement in legume. Seeds of the model legume Lotus japonicus were treated with ethylmethane sulfonate and screening of 40,000 M2 seeds led to the isolation of one mutant named sleepless. sleepless is incapable of closing its leaflets towards the adaxial side at night. The pulvini at the leaflet base were found to be replaced with petiole-like structure in sleepless. Wild-type pulvini comprise many compressed cells, whereas the corresponding region in sleepless is made up of roundish cells in the cortical parenchyma and highly elongated cells in the epidermis, particularly in the leaf-length direction. Based on the results of histological examination, I propose a possible model of a developmental pathway leading to nyctinastic movement. PMID- 12736787 TI - Putative cis-elements in the promoter region of the carrot phenylalanine ammonia lyase gene induced during anthocyanin synthesis. AB - Deletion mutants of the carrot phenylalanine ammonia-lyase gene promoter were used to survey cis-elements for their effect on expression of promoter activity by transient expression. Two putative cis-elements were required to give full activity, but a third might be the most important in regulation of the promoter by 2,4-dichlorophenoxyacetic acid. PMID- 12736788 TI - Expression of novel rice gibberellin 2-oxidase gene is under homeostatic regulation by biologically active gibberellins. AB - We have cloned two genes for gibberellin (GA) 2-oxidase from rice ( Oryza sativa L.). Expression of OsGA2ox2 was not observed. The other gene, OsGA2ox3, was expressed in every tissue examined and was enhanced by the application of biologically active GA. Recombinant OsGA2ox3 protein catalyzed the metabolism of GA(1) to GA(8) and GA(20) to GA(29)-catabolite. These results indicate that OsGA2ox3 is involved in the homeostatic regulation of the endogenous level of biologically active GA in rice. PMID- 12736789 TI - Electrophoretic evidence for homoeologous chromosome pairing in the apogamous fern species Dryopteris nipponensis (Dryopteridaceae). AB - Segregation of genotypes through homoeologous chromosome pairing in the apogamous species Dryopteris nipponensis was tested by electrophoretic analysis. Of 284 progeny examined (250 gametophytes and 34 sporophytes), from the parental sporophyte with the Pgi-2 genotype abc, five showed different genotypes from that of the parent (three aac, one bbc and one bcc). This is the first evidence for genetic segregation in the progeny of apogamous fern species. PMID- 12736790 TI - Population genetics of threatened wild plants in Japan. AB - Approximately one-fourth of Japan's native plant species are threatened with extinction. To conserve these species, it is critical to evaluate genetic diversity at species-level and population-level. Some factors, including population size and geographic distribution, are known to influence the population genetic diversity of wild plant species. This article briefly reviews the population genetic studies that have been conducted on wild threatened plants in Japan. A large population size or wide geographic distribution does not always lead to large genetic diversity, suggesting that historical factors such as speciation processes and population expansion often play more important roles in determining genetic diversity than the number of remnant individuals. The mating system of a species also affects genetic diversity; predominantly selfing species tend to have smaller genetic diversity than outcrossing congeners. Another issue of concern in the conservation genetics of wild plants in Japan is the genetic diversity of insular endemics, because Japan consists of many islands, and the insular flora contains many endemic and threatened species. Previous studies on endemic plants on the Bonin and the Ryukyu Islands are reviewed. Compared to the cases of the Bonin Islands or other oceanic islands, there is much larger genetic diversity in plants endemic to the Ryukyu Islands. This difference is probably the result of the differences in the geological history of these islands. PMID- 12736791 TI - Infantile diarrhoea associated with sorbitol-fermenting, non-shiga toxin producing Escherichia coli O157:H-. PMID- 12736792 TI - Rapid detection of non-multidrug-resistant and multidrug-resistant methicillin resistant Staphylococcus aureus using cycling probe technology for the mecA gene. PMID- 12736793 TI - Shift of the tick Ixodes ricinus and tick-borne encephalitis to higher altitudes in central Europe. PMID- 12736794 TI - Triple Faeces Test: an effective tool for detection of intestinal parasites in routine clinical practice. AB - Microscopic examination of stool specimens is the cornerstone of detection of intestinal parasites in parasitology laboratories. In Europe, fresh, nonpreserved stool specimens are generally used for examination. Because intestinal parasites are shed intermittently, patients are asked to deliver multiple stool samples for examination. The limitation of this diagnostic approach is that detection of the vegetative stages of protozoa may be missed because of delays in processing and/or low compliance with the request to submit multiple stool samples. To overcome this limitation, a diagnostic test that combines multiple sampling (on 3 consecutive days), a fixative (SAF; sodium acetate acetic acid formalin), a concentration method and an easy-to-use permanent stain (chlorazol black dye) was developed for use in routine clinical practice. The results of the test, called the "Triple Faeces Test" (TFT), were compared with those of the conventional diagnostic method, i.e. ether sedimentation of a single fresh stool specimen. Stool samples from 544 patients were examined. Vials from the TFT-sets were filled by patients precisely according to instructions in 462 of 544 (85%) of the cases. Using the conventional method and the TFT, 106 and 209 patients, respectively, were diagnosed with infection by one or more parasitic species ( P<0.005). Pathogenic species were detected by the conventional method and by the TFT in 39 and 94 cases, respectively, and nonpathogenic species were detected in 124 and 288 cases, respectively ( P<0.05). Additional costs for the sampling device, laboratory reagents and handling of the TFT were acceptable. The results of this study suggest that the TFT is an effective method for detection of intestinal parasites in stool samples in routine clinical practice. PMID- 12736797 TI - Visceral leishmaniasis presenting as subcutaneous nodules in a HIV-positive patient. PMID- 12736795 TI - Human Tula virus infection or rat-bite fever? PMID- 12736798 TI - First report of clinical and microbiological failure in the eradication of glycopeptide-intermediate methicillin-resistant Staphylococcus aureus carriage by mupirocin. PMID- 12736799 TI - Identification of Staphylococcus species in blood cultures by a rapid commercial DNA probe test. PMID- 12736800 TI - Representing tools: how two non-human primate species distinguish between the functionally relevant and irrelevant features of a tool. AB - Few studies have examined whether non-human tool-users understand the properties that are relevant for a tool's function. We tested cotton-top tamarins (Saguinus oedipus) and rhesus macaques (Macaca mulatta) on an expectancy violation procedure designed to assess whether these species make distinctions between the functionally relevant and irrelevant features of a tool. Subjects watched an experimenter use a tool to push a grape down a ramp, and then were presented with different displays in which the features of the original tool (shape, color, orientation) were selectively varied. Results indicated that both species looked longer when a newly shaped stick acted on the grape than when a newly colored stick performed the same action, suggesting that both species perceive shape as a more salient transformation than color. In contrast, tamarins, but not rhesus, attended to changes in the tool's orientation. We propose that some non-human primates begin with a predisposition to attend to a tool's shape and, with sufficient experience, develop a more sophisticated understanding of the features that are functionally relevant to tools. PMID- 12736802 TI - Immunoglobulin KM genes in Guillain-Barre syndrome. AB - Guillain-Barre syndrome is associated with antecedent Campylobacter jejuni infection. Only a minority of the infected individuals, however, develops the disease, implying a role for genetic factors in conferring susceptibility. To determine the role of immunoglobulin KM genes (genetic markers of the constant region of kappa chains) in the etiology of this syndrome, we genotyped 83 patients and 196 healthy controls from Norway for KM1 and KM3 alleles by polymerase chain reaction-restriction fragment length polymorphism. The frequency of KM3 homozygotes was significantly increased in patients compared with controls (86.7% vs. 74%, P=0.01, odds ratio=2.3). Conversely, the frequency of KM1/KM3 heterozygotes was significantly decreased in patients compared with controls (13.3% vs. 26%, P=0.01, odds ratio=0.4). These results suggest that KM genes may be relevant to the etiology of Guillain-Barre syndrome. PMID- 12736801 TI - Apolipoproteins E and C1 and brain morphology in memory impaired elders. AB - Previous research has shown that polymorphisms of the apolipoproteins E ( APOE) and APOC1 represent genetic risk factors for dementia and for cognitive impairment in the elderly. The brain mechanisms by which these genetic variations affect behavior or clinical severity are poorly understood. We studied the effect of APOE and APOC1 genes on magnetic resonance imaging measures in a sample of 50 subjects with age-associated memory impairment. The APOE E4 allele was associated with reduced left hippocampal volumes and APOE*E3 status was associated with greater frontal lobe white matter volumes. However, no APOE effects were observed when analyses accounted for other potential confounding variables. The effects of APOC1 on hippocampal volumes appeared to be more robust than those of the APOE polymorphism. However, no modulatory effects on brain morphology outside the medial temporal lobe region were observed when demographic variables, clinical status, and other anatomical brain measurements were taken into consideration. Our results suggest that the role of the APOC1 polymorphism in brain morphology of the cognitively impaired elderly should be examined in further studies. PMID- 12736803 TI - Significant association of the arylalkylamine N-acetyltransferase ( AA-NAT) gene with delayed sleep phase syndrome. AB - Arylalkylamine N-acetyltransferase (AA-NAT) is a rate-limiting enzyme in melatonin hormone synthesis and participates in daily oscillations of the melatonin level. We studied the association between the AA-NAT gene and delayed sleep phase syndrome (DSPS). Results indicate that there is a significant difference in allele positivity at the single nucleotide polymorphism involved in an amino acid substitution from alanine to threonine at position 129 between patients with DSPS and healthy controls. The frequency of the 129 threonine allele is significantly higher in the patients than in the controls ( P=0.0029). The data suggest that AA-NAT could be a susceptibility gene for DSPS. PMID- 12736805 TI - Increased blood pressure in adolescents of low socioeconomic status with short stature. AB - The nutritional programming hypothesis, which has been studied since the 1970s, proposes that intrauterine undernutrition continuing during the first years of life causes permanent metabolic disorders. These alterations are amplified with time, depending on the quality of the diet and on environmental factors. The aim of this cross-sectional study was to detect blood pressure alterations in teenagers with nutritional deficit. The study sample consisted of 53 adolescents living in shantytowns of Sao Paulo City, Brazil; 27 boys and 26 girls ranging in age from 11 to 16 years were studied. Weight, height, skinfold thickness, and blood pressure were measured during a medical visit. Anthropometric data were compared with the reference values of the National Center for Health Statistics and Frisancho, and arterial pressure data were compared with the reference values of the 1996 Task Force. The adolescents studied are among the 10% poorest stratum of the Brazilian population, with a per capita income of U.S. $45/month. The anthropometric results revealed nutritional deficiencies for the entire sample. The major marker of malnutrition was height, with a mean height deficit of 7 cm among boys and of 5 cm among girls. The overall prevalence of arterial (diastolic) hypertension for the sample was 21% (95% confidence interval 10% 32%). No significant difference was observed between sexes. The prevalence of cases with a systolic or diastolic arterial pressure above the 90th percentile, adjusted for height, was 51% ( n=27) (95% confidence interval 37%-65%); 6% ( n=3) of these individuals had simultaneous systolic and diastolic arterial hypertension. In conclusion, the prevalence of arterial hypertension was elevated among malnourished adolescents. The mechanisms involved in the genesis of hypertension seem to be related to malnutrition during an early phase of life, supporting the programming hypothesis. PMID- 12736806 TI - Cerebral aspergillosis in an infant with corticosteroid-resistant nephrotic syndrome. AB - Cerebral aspergillosis is a devastating disease in patients with a compromised immune system. A unique case of a male infant with corticosteroid-resistant nephrotic syndrome complicated by pulmonary and cerebral aspergillosis is described. The patient rapidly developed coma and neurological symptoms and died soon thereafter. Central nervous system aspergillosis was diagnosed radiologically and by detection of Aspergillus DNA and antigen (galactomannan) in blood and cerebrospinal fluid. Moderate immunosuppressive therapy and antifungal phagocytic dysfunction due to nephrotic syndrome per se may have contributed to the occurrence of invasive aspergillosis in our patient. Awareness of this serious complication and early application of diagnostic procedures and antifungal therapy may improve the dismal outcome. PMID- 12736807 TI - Autoimmune lymphoproliferative syndrome presenting with glomerulonephritis. AB - Autoimmune lymphoproliferative syndrome (ALPS) is characterized clinically by chronic non-malignant lymphoproliferation and autoimmunity and is caused by a genetic defect in programmed cell death (apoptosis). Most patients with ALPS have heterozygous mutations in the Fas gene. We describe an 11-year-old Brazilian boy with hepatosplenomegaly, lymphadenopathy, hemolytic anemia, and hypergammaglobulinemia since early infancy. T cell lines from the patient were defective in Fas-mediated apoptosis. He was diagnosed as having ALPS and found to have a novel Fas gene mutation (IVS4+1G>A). In addition, he presented with glomerulonephritis in infancy. An aunt and uncle who had the same Fas mutations also had histories of glomerulonephritis. Although glomerulonephritis is common in Fas-deficient mice, it is infrequent in human ALPS. Corticosteroid therapy ameliorated the glomerulonephritis in our patient, as well as his lymphoproliferation, anemia, and hypergammaglobulinemia. This study suggests that glomerulonephritis is one of the characteristic features of ALPS. PMID- 12736808 TI - Adenovirus pyelonephritis in a pediatric renal transplant patient. AB - Gross hematuria, graft pain, and rising serum creatinine are classic signs of acute rejection, obstruction, or bacterial pyelonephritis for patients with renal transplants. This presentation often prompts percutaneous renal allograft biopsy. If subsequent evaluation fails to show evidence of acute rejection, obstruction, or bacterial infection, viral etiologies should be considered. We report a 14 year-old Hispanic female with a living-related renal transplant who had gross hematuria, graft tenderness, and increased serum creatinine, but did not have evidence of acute rejection, obstruction, or bacterial pyelonephritis. To our knowledge, this is the first report of adenovirus pyelonephritis in a transplanted kidney of a pediatric patient, with isolation of adenovirus in the urine and in the allograft using immunocytochemical techniques. PMID- 12736809 TI - Mild rhabdomyolysis in a child with fever and "hematuria". AB - Rhabdomyolysis represents a life-threatening condition, which results in release of cellular contents (myoglobin, enzymes, and electrolytes) into the plasma. We report a pediatric patient with mild rhabdomyolysis who had a favorable outcome. A 3-year-old girl had been ill for 2 days with high fever, anorexia, pain in both thighs, and passage of dark-red urine. Myoglobinuria was demonstrated by a "blood"-positive dipstick in the absence of red blood cells in the urinary sediment. Diagnosis was confirmed by the presence of a high serum creatine kinase activity. The child was treated on an outpatient basis and has shown full clinical and biochemical recovery. There has been no recurrence of myoglobinuria during the 2-year follow-up. PMID- 12736810 TI - Peripheral gangrene complicating hemolytic uremic syndrome in a child. AB - A patient with hemolytic uremic syndrome (HUS) developed peripheral gangrene involving all fingers and toes. There was no history of bloody diarrhea. Hypocomplementemia was present, with a serum C3 concentration of 41 mg/dl. Acute renal failure was treated with peritoneal dialysis for 4 months. He received daily fresh-frozen plasma infusions and plasmapheresis on alternate days for ten sessions, followed by once-weekly sessions. He was anuric for 9 weeks. All medial and distal phalanxes became necrotic and were removed surgically. The renal biopsy findings were consistent with HUS. This is the second report of peripheral gangrene during the course of HUS in childhood. PMID- 12736811 TI - Antibiotic prophylaxis by low-dose cefaclor in children with vesicoureteral reflux. AB - Sulfamethoxazole/trimethoprim (SMX/TMP) and nitrofurantoin are the most frequently used agents for prophylaxis to reduce the risk of recurrent urinary tract infections (UTIs) in children with vesicoureteral reflux (VUR). Nitrofurantoin, however, is not available in Japan and increasing resistance of organisms to SMX/TMP has recently raised doubts about its effectiveness as a prophylactic agent. This study was conducted to investigate whether antibiotic prophylaxis using low-dose cefaclor can effectively reduce the risk of recurrent UTIs. Thirty-nine children (31 male, 8 female) with primary VUR were enrolled. Ages varied from 0.5 to 111 months (mean 10.6 months). A prophylactic dose of 5 10 mg cefaclor per kg per day was given 1-3 times daily depending on the patient's age. Mean duration of prophylactic treatment was 15.5 months. Eleven children (ten male, one female) developed breakthrough UTIs during a total of 606 months treatment (or about one further infection in 55 months). Resistance to cefaclor was noted in three organisms: Enterococcus spp., Morganella spp., and Pseudomonas spp. Evidence of antibacterial activity was present in the morning urine samples from all of seven children tested. Cefaclor was well accepted and tolerated by all subjects. None withdrew from the study because of side effects. These results suggest that cefaclor can be an alternative choice for prophylactic treatment because of its safety, good compliance and low rates of resistant Escherichia coli. PMID- 12736813 TI - Reversible secondary pseudohypoaldosteronism. PMID- 12736812 TI - Renal biopsy 2-9 years after Henoch Schonlein purpura. AB - Twelve children and adolescents with classical Henoch-Schonlein purpura (HSP) underwent renal biopsy 2-9 years later. The clinical course was favorable in 10 of the patients who received only supportive treatment. Two patients with a prolonged course, characterized by marked hematuria and proteinuria, were given steroids and azathioprine first and mycophenolate mofetil later. At the time of biopsy, 4 patients did not have any urinary abnormalities, 6 had hematuria and/or mild proteinuria, and 2 had proteinuria >1 g/24 h. Renal histology showed mild lesions on light microscopy. Immunohistochemistry disclosed mesangial IgA in 8 of the 12 patients. In percentages, this is a proportion of 66.6 with 95% exact confidence limits of 34.9-90.1. In this small series of selected former HSP patients, the majority had IgA nephropathy years after the initial vasculitis episode. This indicates that some patients with apparently completely healed HSP have a chronic glomerular condition that possibly means protracted disease and certainly indicates the need for careful follow-up. PMID- 12736814 TI - Endoscopic approach for carotid artery surgery (018214). PMID- 12736815 TI - Should we forget the road that led to where we are now? PMID- 12736817 TI - Chromosome analysis of Phyllodistomum folium (Trematoda, Gorgoderidae) infecting three European populations of zebra mussels. AB - The mitotic chromosome sets of the larval stages of Phyllodistomum folium infecting three European populations of Dreissena polymorpha were studied using conventional Giemsa staining and karyometrical analysis. The karyotype, described herein for the first time, consisted of nine chromosome pairs (2 n=18). The chromosomes were comparatively small and measured from 1.19 micro m to 5.22 micro m. The first and second longest chromosome pairs were, respectively, approximately 20% and 18% of the mean total chromosome set length and were clearly differentiated from the other seven pairs, which gradually decreased in size. Most chromosomes were biarmed with medially or submedially located centromeres; only pairs 1 and 4 contained subterminally located centromeres. No significant differences in the relative length values of corresponding chromosomes were observed between the three populations studied. The main interpopulation differences were detected in the centromeric index values of some corresponding chromosome pairs. These data, when analysed in conjunction with those of other Gorgoderidae, indicate that the karyotype of P. folium and other species in this family do not show any clear affinities with "typical" plagiorchiate chromosome sets. PMID- 12736816 TI - Common 5' beta-globin RFLP haplotypes harbour a surprising level of ancestral sequence mosaicism. AB - Blocks of linkage disequilibrium (LD) in the human genome represent segments of ancestral chromosomes. To investigate the relationship between LD and genealogy, we analysed diversity associated with restriction fragment length polymorphism (RFLP) haplotypes of the 5' beta-globin gene complex. Genealogical analyses were based on sequence alleles that spanned a 12.2-kb interval, covering 3.1 kb around the psibeta gene and 6.2 kb of the delta-globin gene and its 5' flanking sequence known as the R/T region. Diversity was sampled from a Kenyan Luo population where recent malarial selection has contributed to substantial LD. A single common sequence allele spanning the 12.2-kb interval exclusively identified the ancestral chromosome bearing the "Bantu" beta(s) (sickle-cell) RFLP haplotype. Other common 5' RFLP haplotypes comprised interspersed segments from multiple ancestral chromosomes. Nucleotide diversity was similar between psibeta and R/T delta-globin but was non-uniformly distributed within the R/T-delta-globin region. High diversity associated with the 5' R/T identified two ancestral lineages that probably date back more than 2 million years. Within this genealogy, variation has been introduced into the 3' R/T by gene conversion from other ancestral chromosomes. Diversity in delta-globin was found to lead through parts of the main genealogy but to coalesce in a more recent ancestor. The well known recombination hotspot is clearly restricted to the region 3' of delta globin. Our analyses show that, whereas one common haplotype in a block of high LD represents a long segment from a single ancestral chromosome, others are mosaics of short segments from multiple ancestors related in genealogies of unsuspected complexity. PMID- 12736818 TI - Depressed interleukin-12 production by peripheral blood mononuclear cells after in vitro stimulation with the 30-kDa antigen in recurrent pulmonary tuberculosis patients. AB - Some patients develop recurrent tuberculosis (R-TB), even after successfully completing initial anti-tubercular treatment. Although R-TB may be caused by relapse or exogenous reinfection, little is known about the underlying host responses associated with R-TB. This study investigated the profile of cytokines [interferon (IFN)-gamma, interleukin (IL)-12, tumor necrosis factor (TNF)-alpha, IL-6, and IL-10] present in peripheral blood mononuclear cells (PBMCs) of 17 R-TB patients after stimulation with the 30-kDa antigen (Ag) or purified protein derivative (PPD) Ag of Mycobacterium tuberculosis. These data were compared with data obtained from 15 patients with newly diagnosed pulmonary TB (N-TB), 22 patients with treatment failure (TF-TB), and 19 healthy tuberculin reactors (HTR). N-TB and R-TB patients were enrolled in this study within 1 month of beginning anti-tubercular chemotherapy. ELISA results showed that IFN-gamma production following stimulation with the 30-kDa Ag was significantly lower in each group of TB patients than in the HTR controls. In addition, patients with R TB showed the most significant IL-12 depression among the subject groups after in vitro stimulation with either Ag. Furthermore, a significant decrease in TNF alpha and IL-10 levels was observed in R-TB patients relative to N-TB patients. However, there was no statistical difference in TNF-alpha and IL-10 production between R-TB patients, TF-TB patients, and HTR controls. Our findings suggest that the underlying mechanisms of cytokine regulation might differ between N-TB and R-TB patients, and that decreased IL-12 production in response to the 30-kDa or PPD Ag might be involved in the immunopathogenesis of human R-TB. PMID- 12736819 TI - Antifungal activity of macrophages engineered to produce IFNgamma: inducibility by picolinic acid. AB - Macrophages are important antimicrobial effectors, whose efficacy is greatly enhanced by interferon-gamma (IFNgamma). We recently engineered a mouse macrophage cell line to express the IFNgamma gene in a inducible manner. Such macrophages, Mphi10, include a construct containing the IFNgamma gene under the control of the synthetic promoter HRE3x-Tk. Picolinic acid (PA) is a catabolite of tryptophan, known to exert costimulatory activities on macrophages and expected to act on transcriptional elements within HRE3x-Tk promoter. Since evidence exists on the efficacy of engineered macrophages as carriers of therapeutic genes against tumors, we tested Mphi10, under basal conditions and following exposure to PA, as IFNgamma-producing cells in in vitro models of fungal infection. We found that Mphi10 constitutively exhibited anticryptococcal and anticandidal activity, low but detectable levels of IFNgamma mRNA and undetectable levels of nitric oxide synthase (iNOS) transcripts. Treatment with PA caused time-dependent enhancement of antifungal activity. The phenomenon was associated with the induction of both IFNgamma and iNOS gene expression and was followed by IFNgamma and NO production. The effect of the Mphi10-produced IFNgamma on other cells was also investigated by a transwell co-culture system. A major enhancement of phagocytosis and antifungal activity was observed in BV2 microglial cells that had been co-cultured with Mphi10. Such an increase was only evident when Mphi10 had been pretreated with PA and was abrogated by concomitant addition of anti-IFNgamma antibodies. In conclusion, we show that Mphi10 respond to PA with the production of IFNgamma, which retains the ability to induce antifungal activity in the producing macrophages as well as in other macrophage populations. The potential use of Mphi10 as vectors for therapeutic genes in infectious diseases is discussed. PMID- 12736821 TI - A Listeria adhesion protein-deficient Listeria monocytogenes strain shows reduced adhesion primarily to intestinal cell lines. AB - Listeria monocytogenes adheres and penetrates intestinal cell linings for systemic infection. A 104-kDa Listeria adhesion protein (LAP) from L. monocytogenes was previously demonstrated to be responsible for adhesion to intestinal enterocyte-like Caco-2 cells. We investigated the adhesion and invasion characteristics of a LAP-deficient mutant L. monocytogenes strain (A572) to various human intestinal and non-intestinal cell lines to assess the possible target host cells. Among the intestinal cell lines, A572 showed significantly reduced adhesion than the wild type (WT) strain to the cells of ileum-cecum (HCT 8) and colon (Caco-2 and HT-29), whereas A572 and WT did not show any significant differences in adhesion to other intestinal cell lines from duodenum (HuTu-80) or jejunum (Int-407). Differences in adhesion between A572 and WT were little or none in non-intestinal cell lines from liver, kidney, bladder, ovary, cervix, breast, larynx, or skin. Invasion data showed that A572 was invasive but the invasion efficiency was proportional to its adhesion characteristics to respective cell lines. In mouse bioassay, A572 was not found in liver following oral administration, suggesting that LAP mutant was possibly unable to pass through intestinal cell linings. Immuno-electron microscopy revealed that the LAP is localized in the bacterial surface as well as the cytoplasm. In summary, this study indicated that the LAP-mediated adhesion is associated with the intestinal cells originating from the lower part of small intestine and from the upper part of large intestine, and possibly plays an important role during the intestinal phase of infection. PMID- 12736820 TI - Differentiation of Borrelia burgdorferi sensu lato strains using class I lysyl tRNA synthetase-encoding genes. AB - The essential protein lysyl-tRNA synthetase (LysRS) exists in two unrelated forms, a class I and a class II-type aminoacyl-tRNA synthetase. Comparative genome sequence analysis revealed that Borrelia burgdorferi sensu lato, the etiological agent of Lyme disease, contains a class I-type LysRS, whereas its tick and mammalian hosts would be expected to contain a class II-type protein. To investigate the utility of the class I LysRS as a diagnostic target for Lyme disease, the corresponding gene ( lysK) was cloned and sequenced from B. afzelii, B. garinii, and B. hermsii. These lysK sequences were then used to design a primer set that could detect and genotype B. burgdorferisensu strictu, B. afzelii, and B. garinii in one single polymerase chain reaction, while showing no cross reactivity with examples of other Borrelia or spirochetes. PMID- 12736822 TI - Immunity against diphtheria and tetanus in German blood donors. AB - After the recent diphtheria epidemics in Eastern Europe in the early 1990s, we re evaluated the diphtheria and tetanus immunity of 321 German blood donors (192 men and 129 women). The mean antitoxin levels of all blood donors in this study, measured by commercial ELISA, revealed a questionable protection (0.1-1.0 IU/ml) against diphtheria. In 1994, 66.4% were without immunity against diphtheria (55.0% in 1997/98), 32.1% (41.5% in 1997/98) showed questionable protection and only 1.5% (3.5% in 1997/98) had protective antitoxin levels. The evaluation of tetanus immunity revealed only 0.5% (1.1% in 1997/98) of the subjects with no protection and 9.1% (8.5% in 1997/98) with questionable protection. For this reason, we conclude that the diphtheria epidemics only lead to an insufficient improvement of the immunization status in a healthy German population. PMID- 12736823 TI - Effective macrophage redox defense against Chlamydia pneumoniae depends on L-type Ca2+ channel activation. AB - Macrophage immune capability depends on their efficient redox potential expressed in the effective release of reactive oxygen species (ROS) and nitric oxide. In this study the effect of the activation of a specialized Ca(2+) channel on macrophage redox function during Chlamydia pneumoniae infection was explored. C. pneumoniae exhibited a profound and sustained Ca(2+) influx capacity, with evidence of activity attributable to their lipopolysaccharide (cLPS) content. Also the organism showed an additional Ca(2+) influx signal in macrophages exposed to thapsigargin, and there was evidence for the operation of a single ion channel of the L type as demonstrated by the effect of L-type channel antagonists (methoxyverapamil and nimodipine) despite exposure to Ca(2+)-rich medium. C. pneumoniae or cLPS induced intracellular ROS and NO generation in a manner consistent with dependence on intracellular calcium. L-type Ca(2+) channel blocking significantly prompted C. pneumoniae inclusion formation. These findings suggest that Ca(2+) influx signal and redox function in C. pneumoniae-infected macrophages depend on L-type Ca(2+) channel activation. PMID- 12736824 TI - A genomewide survey of developmentally relevant genes in Ciona intestinalis. I. Genes for bHLH transcription factors. AB - The basic helix-loop-helix (bHLH) proteins are transcription factors that play important roles in many biological processes, including the development of various animals. We identified 46 genes encoding bHLH proteins in the draft genome sequence of the basal chordate Ciona intestinalis. These 46 genes represent an almost complete set of bHLH genes in this animal. This number is comparable to 39 and 58 bHLH genes in those of Caenorhabditis elegans and Drosophila melanogaster but is much smaller than the 125 in the human genome. Genes that encode NeuroD, Beta3, Oligo, SCL, NSCL, SRC, Clock and Bmal were not found in the Ciona genome. We could also identify groucho and CBP orthologues in the Ciona genome. In addition, the comparison between the numbers of orthologous human and Ciona bHLH factors reveals the simplicity of the Ciona genome, in accordance with its phylogenetic position within chordates. The present analysis provides an insight into the basic set of bHLH genes that the last common ancestor of ascidians and vertebrates had, and will provide important background information for future studies using ascidians as a model system for vertebrate development. PMID- 12736825 TI - A genomewide survey of developmentally relevant genes in Ciona intestinalis. II. Genes for homeobox transcription factors. AB - Homeobox-containing genes play crucial roles in various developmental processes, including body-plan specification, pattern formation and cell-type specification. The present study searched the draft genome sequence and cDNA/EST database of the basal chordate Ciona intestinalis to identify 83 homeobox-containing genes in this animal. This number of homeobox genes in the Ciona genome is smaller than that in the Caenorhabditis elegans, Drosophila melanogaster, human and mouse genomes. Of the 83 genes, 76 have possible human orthologues and 7 may be unique to Ciona. The ascidian homeobox genes were classified into 11 classes, including Hox class, NK class, Paired class, POU class, LIM class, TALE class, SIX class, Prox class, Cut class, ZFH class and HNF1 class, according to the classification scheme devised for known homeobox genes. As to the Hox cluster, the Ciona genome contains single copies of each of the paralogous groups, suggesting that there is a single Hox cluster, if any, but genes orthologous to Hox7, 8, 9 and 11 were not found in the genome. In addition, loss of genes had occurred independently in the Ciona lineage and was noticed in Gbx of the EHGbox subclass, Sax, NK3, Vax and vent of the NK class, Cart, Og9, Anf and Mix of the Paired class, POU-I, III, V and VI of the POU class, Lhx6/7 of the LIM class, TGIF of the TALE class, Cux and SATB of the Cut class, and ZFH1 of the ZFH class, which might have reduced the number of Ciona homeobox genes. Interestingly, one of the newly identified Ciona intestinalis genes and its vertebrate counterparts constitute a novel subclass of HNF1 class homeobox genes. Furthermore, evidence for the gene structures and expression of 54 of the 83 homeobox genes was provided by analysis of ESTs, suggesting that cDNAs for these 54 genes are available. The present data thus reveal the repertoire of homeodomain-containing transcription factors in the Ciona genome, which will be useful for future research on the development and evolution of chordates. PMID- 12736826 TI - Large scale EST analyses in Ciona intestinalis: its application as Northern blot analyses. AB - We have conducted large-scale EST analyses of transcripts expressed in the basal chordate Ciona intestinalis. The cDNA libraries examined were from fertilized eggs, cleaving embryos, gastrulae/neurulae, tailbud embryos, larvae and whole young adults, and the gonad (ovary and testis), endostyle, neural complex, heart, and blood cells of the adult. Because the libraries were not normalized or amplified, the occurrence of cDNA clones or EST counts in each library may reflect the quantity of transcripts of the corresponding genes. Thus, comparison of the EST counts of a certain gene at the six developmental stages may reflect the temporal expression pattern of the gene, while the comparison of EST counts in different tissues of the adult may reflect the spatial expression pattern of the gene. Here we present evidence that this is the case, and that the EST count can therefore be used like "Northern blot analysis" of Ciona genes. PMID- 12736827 TI - Genomewide surveys of developmentally relevant genes in Ciona intestinalis. PMID- 12736828 TI - The expression of tissue inhibitor of metalloproteinase 2 (TIMP-2) is required for normal development of zebrafish embryos. AB - MMP activities are controlled by a combination of proteolytic pro-enzyme activation steps and inhibition by endogenous inhibitors like alpha2 macroglobulin and the tissue inhibitors of metalloproteinases (TIMPs). TIMPs are the key inhibitors in tissue. The expression of both MMPs and TIMPs is controlled during tissue remodeling to maintain a balance in the turnover of extracellular matrix. Disruption of this balance may result in a broad spectrum of diseases. Additionally, TIMP-2 has been reported to have growth factor activities. To further study the function of TIMP-2 in development, we utilized zebrafish as an experimental model system. We have successfully isolated a TIMP-2 homologue from zebrafish (zTIMP-2). This zebrafish TIMP-2 showed high similarity to human TIMP-2 with all critical features conserved. Whole-mount in situ analysis showed that zTIMP-2 was expressed as early as the one-cell stage indicating a maternal origin. This expression continued through later stages of development. RT-PCR analysis confirmed the early expression pattern from the 16-cell stage through blastula, gastrula and 24-h stages. In addition, at the protein level, immunoreactive zTIMP-2 was detected using antibody against recombinant human TIMP 2. RFP-reporter analysis indicated that TIMP-2 can be secreted into the extracellular space where ECM is forming. Functional studies showed that the balance of TIMP-2 expression is important to normal development as reflected by the fact that both blockage of TIMP-2 translation using antisense morpholino oligonculeotides or increased translation of TIMP-2 using a mRNA microinjection approach resulted in abnormal zebrafish development. This is in contrast to murine knockout studies that indicate that TIMP-2 does not have a major role in mouse embryogenesis. PMID- 12736830 TI - Complexity analysis of the temperature curve: new information from body temperature. AB - An attempt was made to develop a truly quantitative approach to temperature, based on models derived from nonlinear dynamics and chaos theory. Three different procedures for measuring the degree of complexity of the temperature curve were compared, and the possible correlations between these measurements and certain physiopathologically relevant parameters in healthy subjects were examined. Twenty-three healthy subjects (10 males, 13 females) between 18 and 85 years of age had their temperature measured every 10 min for at least 30 h. These time series were used to determine the approximate entropy (ApEn), a detrended fluctuation analysis (DFA), and the fractal dimension by the compass method (FD(c)). There was good correlation between the different methods of measuring the complexity of the curve [ r=-0.603 for ApEn vs. DFA ( p=0.002), r=0.438 for ApEn vs. FDc ( p=0.04) and r=-0.647 for DFA vs. FDc ( p=0.0008)]. Both the fractal dimension and the approximate entropy were inversely correlated with age [ r=-0.637 ( p=0.001) and r=-0.417 ( p=0.03), respectively], while the DFA increased with age ( r=0.413, p=0.04). The results thus suggest that complexity of the temperature curve decreases with age. The complexity of the temperature curve can be quantified in a consistent fashion. Age is associated with lower complexity of the temperature curve. PMID- 12736829 TI - Comparison of cardio-locomotor synchronization during running and cycling. AB - By comparing the characteristics of cardiac-locomotor synchronization (CLS) in running and cycling individuals, we tested whether the characteristics of CLS occurring during rhythmic exercise adhere to the central origin hypothesis, which postulates a direct interaction between cardiovascular centers in the brain and the pattern generator in the spinal cord. Ten healthy subjects performed both exercises at the same intensity (150 beats.min(-1)) and cadence (150 steps.min( 1) during running and 75 rpm during cycling), while electrocardiograms and electromyograms from the right vastus lateralis muscle were monitored continuously. An examination of the occurrence of heart beats with respect to the locomotor phase revealed that, in running subjects, CLS exists for relatively prolonged periods at specific phases, whereas, in cycling subjects, it occurs intermittently and is not phase-specific [maximum duration of CLS: 113.6 (66.5) and 58.0 (29.3) s ( P<0.05), respectively]. Determining the probability of CLS by chance as a function of its duration, we also found that, during running, CLS likely results from entrainment, whereas, during cycling, it results from chance, occurring when the cardiac rhythm approached the locomotor rhythm. Our result indicated that the duration of muscle contraction during cycling [317.0 (18.1) ms] was significantly longer than during running [205.6 (20.2) ms]. These results indicated that the difference in the CLS characteristics between running and cycling might be influenced by differences in peripheral inputs between exercise modes. PMID- 12736832 TI - Reduced length-dependent cross-bridge recruitment in skinned fiber preparations of human failing myocardium. AB - A depressed activity of myosin ATPase has been described in human failing myocardium. Since alterations in cross-bridge kinetics may affect both systolic and diastolic cardiac function, the present study simultaneously investigated Ca(2+)-dependent tension and actomyosin ATPase activity (MYO) in triton X-skinned fiber preparations of human non-failing (donor hearts, n=8) and failing (dilated cardiomyopathy, n=11) left ventricular myocardium at increasing sarcomeric length (1.9 and 2.1 microm, alpha-actinin staining). The MYO/tension ratio was analyzed as a parameter characterizing myofibrillar energetics. At a sarcomere length of 1.9 microm, the Ca(2+) sensitivity of tension was significantly increased in human failing compared to non-failing myocardium. In human non-failing myocardium, maximal Ca(2+)-activated tension [1.9 microm vs. 2.1 microm, 23.7 (1.9) vs. 28.3 (1.9) mN/mm(2)] and the Ca(2+) sensitivity of tension [EC(50)Ca(2+ )(pCa): 5.67 (0.06) vs. 7.07 (0.11)] were increased by increasing sarcomere length. This was accompanied by an enhancement in Ca(2+)-dependent MYO [+72 (11) vs. +101 (9) microM ADP/s] as well as an increase in the Ca(2+)-sensitivity of MYO [EC(50)Ca(2+ )(pCa): 5.84 (0.08) vs. 6.86 (0.08)]. In human failing myocardium, only Ca(2+) sensitivity of tension (but not of MYO) increased. Tension cost was increased in failing vs. non-failing tissue [1.9 microm: 4.18 (0.06) vs. 3.53 (0.06) (mN.s)/(mm(2). microM ADP); 2.1 microm: 4.28 (0.13) vs. 3.52 (0.05) (mN.s)/(mm(2). microM ADP)]. We concluded that, in human failing myocardium, the length-dependent force generation may be blunted due to an already increased Ca(2+) affinity of troponin C as well as an impairment of length-dependent cross-bridge recruitment. PMID- 12736831 TI - Growth trend of the quadriceps femoris muscle in junior Olympic weight lifters: an 18-month follow-up survey. AB - Isometric torque and the cross-sectional area (CSA) of the quadriceps femoris muscle (QF) were measured twice at an interval of 18 months in seven junior Olympic weight lifters (aged 15.5-17.1 years at the start of the study) to investigate the growth trend of this muscle group in junior athletes specializing in strength and power events. The CSAs of the QF and its four constituent muscles were determined at 30% (proximal to the knee), 50%, and 70% of femur length (Fl) using a magnetic resonance imaging method. Only at 30%Fl were significant increases found in the CSAs of the QF and all constituent muscles, except for the rectus femoris. At this level, the CSA of the vastus medialis relative to the QF decreased significantly, suggesting a predominant hypertrophy in the vastus intermedius and vastus lateralis muscles. Isometric torque during knee extension increased significantly compared, to the CSA for the QF measured at the three levels. Thus, the present results indicate that, at least at high school age, the QF for junior Olympic weight lifters is characterized by a preferential hypertrophy at the level proximal to the knee joint, with an increase in knee extension torque relative to the CSA. PMID- 12736833 TI - Effect of exercise to exhaustion on myeloperoxidase and lysozyme release from blood neutrophils. AB - Exercise sessions (swimming in rats and treadmill running in humans) resulted in stimulation of neutrophil degranulation in the experiments with animals and in the human study. Myeloperoxidase (MPO) (+67%) and lysozyme (+51%) quantities in the plasma of rats increased significantly immediately after exercise. The blood plasma lysozyme concentration was increased by 41% at the 6th min of treadmill exercise in athletes. The blood concentrations of neutrophil proteins normalized both in humans and animals at rest. The neutrophil protein concentrations in blood increased in parallel with the decrease of their level in leukocytes. The neutrophil capacity for an oxidative burst was not changed by the exercise, but decreased for 3-6 h in the post-exercise period. Such dynamics of the oxidative burst activity suggest a lack of association between this parameter and the degranulation process. The neutrophil proteins that appear in blood during degranulation can be involved in enhancing the bactericidal potency of blood, the activation of granulopoiesis, neutrophil efflux from bone marrow, and the conditioning of blood endothelium for leukocyte extravasation. PMID- 12736834 TI - Training mode does not affect orthostatic tolerance in chronically exercising subjects. AB - This study tested the hypotheses that trained swimmers would have greater orthostatic tolerance than runners and, if present, it would be due to differences in their autonomic and hemodynamic responses to graded central hypovolemia. Twenty intercollegiate male athletes [11 runners and 9 swimmers; .VO(2max) =70.0 (1.6) vs 69.5 (2.6) ml.kg(-1).min(-1), respectively] underwent graded lower body negative pressure (LBNP) to presyncope. The swimmers were heavier [80.5 (1.9) vs 70.3 (1.9) kg, P<0.05], with larger resting cardiac [4.44 (0.29) vs 3.68 (0.18) l.min(-1).m(-2)] and total peripheral conductance [0.056 (0.04) vs 0.044 (0.02) units.m(-2)] indices. Neither spontaneous cardiac baroreflex sensitivity (sequence method) nor heart rate variability (spectral analysis) differed significantly between groups at rest. LBNP tolerance did not differ between groups, with an index value of 51 (2) kPa.min for the runners and 54 (4) kPa.min for the swimmers [383 (16) vs 402 (32) mmHg.min], although the swimmers had larger declines in pulse pressure and tended ( P=0.078) to have larger declines in total peripheral conductance index in the last completed stage of LBNP. These responses did not differ between groups in the last 2 min of LBNP. Neither the heart rate, mean arterial pressure nor forearm vascular conductance responses differed between groups throughout. Changes in heart rate variability indices did not differ significantly between groups, with similar declines in the high frequency component and increases in the low frequency/high frequency ratio. These data suggest that swim training does not lead to greater orthostatic tolerance than run training, and responses to maximal LBNP do not differ between swimmers and runners. Moreover, neither heart rate nor the autonomic modulation of the heart rate response to LBNP are affected by training modality. PMID- 12736835 TI - Antagonist moment of force during maximal knee extension in pubertal boys: effects of quadriceps fatigue. AB - The examination of the moment exerted by the hamstrings during maximum isokinetic knee extensor tests is useful when comparing isokinetic strength and muscle activity patterns between children and adults. The purpose of this study was to examine the effect of antagonist moment of the hamstrings on the isokinetic moment of the knee extensors in pubertal children and to determine whether this effect is altered following a fatigue task. Eighteen healthy pubertal males [age 14.3 (0.5) years] performed 34 maximal isokinetic concentric efforts of the knee extensors at 60 degrees.s(-1). The average moment of force and electromyographic (aEMG) signal of vastus medialis (VM), vastus lateralis (VL) and biceps femoris (BF) at 11-30 degrees, 31-50 degrees, 51-70 degrees and 71-90 degrees of knee flexion were calculated for each repetition. The hamstrings antagonist moment was determined before and after the fatigue task by fitting the aEMG-moment relationship at different levels of muscle effort using second-degree polynomials. The percentage contribution of the antagonist moment to the resultant joint moment ranged from 7.1 % to 60.4 % throughout the range of motion, with the highest percentage observed close to full knee extension (11-30 degrees). The antagonist effect was significantly greater during concentric tests of the knee extensors compared to the corresponding eccentric tests ( p<0.05). Following the fatigue test, there was an overall decline of the resultant joint moment, but no changes in the predicted hamstrings moment were observed. These results indicate that when testing maximal knee extensor isokinetic strength in pubertal boys, activity of the hamstrings implies a reduction of the net extensor moment as compared to the isolated capacity of the knee extensors. However, this antagonist effect is not altered following the performance of an isokinetic fatigue knee extension task. PMID- 12736836 TI - Maximal lactate steady state, respiratory compensation threshold and critical power. AB - Critical power (CP) and the second ventilatory threshold (VT(2)) are presumed to indicate the power corresponding to maximal lactate steady state (MLSS). The aim of this study was to investigate the use of CP and VT(2) as indicators of MLSS. Eleven male trained subjects [mean (SD) age 23 (2.9) years] performed an incremental test (25 W.min(-1)) to determine maximal oxygen uptake (.VO(2max)), maximal aerobic power (MAP) and the first and second ventilatory thresholds (VT(1) and VT(2)) associated with break points in minute ventilation (.V(E)), carbon dioxide production (.VCO(2)), .V(E)/.VCO(2) and .V(E)/.VO(2) relationships. Exhaustion tests at 90%, 95%, 100% and 110% of .VO(2max), and several 30-min constant work rates were performed in order to determine CP and MLSS, respectively. MAP and .VO(2max) values were 344 (29) W and 53.4 (3.7) ml.min(-1).kg(-1), respectively. CP [278 (22) W; 85.4 (4.8)% .VO(2max)] and VT(2) power output [286 (28) W; 85.3 (5.6)% .VO(2max)] were not significantly different (p=0.96) but were higher (p<0.05) than the MLSS work rate [239 (21) W; 74.3 (4.0)% .VO(2max)] and VT(1) power output [159 (23) W; 52.9 (6.9)% .VO(2max)]. MLSS work rate was significantly correlated (p<0.05) with those noted at VT(1) and VT(2) (r=0.74 and r=0.93, respectively). VT(2) overestimated MLSS by 10.9 (6.3)% .VO(2max), which was significantly higher than VT(1) [+21.4 (5.6)% .VO(2max); p<0.01]. CP calculated from a given range of exhaustion times does not correspond to MLSS. PMID- 12736837 TI - Oxygen uptake kinetics during moderate, heavy and severe intensity "submaximal" exercise in humans: the influence of muscle fibre type and capillarisation. AB - The purpose of the present study was to test the hypothesis that muscle fibre type influences the oxygen uptake (.VO(2)) on-kinetic response (primary time constant; primary and slow component amplitudes) during moderate, heavy and severe intensity sub-maximal cycle exercise. Fourteen subjects [10 males, mean (SD) age 25 (4) years; mass 72.6 (3.9) kg; .VO(2peak) 47.9 (2.3) ml kg(-1) min( 1)] volunteered to participate in this study. The subjects underwent a muscle biopsy of the vastus lateralis for histochemical determination of muscle fibre type, and completed repeat "square-wave" transitions from unloaded cycling to power outputs corresponding to 80% of the ventilatory threshold (VT; moderate exercise), 50% (heavy exercise) and 70% (severe exercise) of the difference between the VT and .VO(2peak). Pulmonary .VO(2) was measured breath-by-breath. The percentage of type I fibres was significantly correlated with the time constant of the primary .VO(2) response for heavy exercise (r=-0.68). Furthermore, the percentage of type I muscle fibres was significantly correlated with the gain of the .VO(2) primary component for moderate (r=0.65), heavy (r=0.57) and severe (r=0.57) exercise, and with the relative amplitude of the .VO(2) slow component for heavy (r=-0.74) and severe (r=-0.64) exercise. The influence of muscle fibre type on the .VO(2) on-kinetic response persisted when differences in aerobic fitness and muscle capillarity were accounted for. This study demonstrates that muscle fibre type is significantly related to both the speed and the amplitudes of the .VO(2) response at the onset of constant-load sub maximal exercise. Differences in contraction efficiency and oxidative enzyme activity between type I and type II muscle fibres may be responsible for the differences observed. PMID- 12736838 TI - Evaluation of the walking pattern in two types of patients with anterior cruciate ligament deficiency: copers and non-copers. AB - The purpose of the present study was to investigate whether different walking patterns in healthy subjects and in coper and non-coper subjects with deficient anterior cruciate ligaments could be quantified. An inverse dynamics approach was used to calculate joint kinematics and kinetics for flexion and extension. EMG signals of the hamstrings and quadriceps muscles were recorded. The results showed that the peak knee flexion angle was greater in the copers than in the controls. There was a positive correlation between the peak knee extensor moment and peak knee flexion angle. Furthermore, at a given peak knee flexion angle, the peak knee extensor moment was significantly larger in the controls than in the non-copers. The hip extensor moment in the copers was significantly larger than that of the non-copers and the controls. In conclusion, the three groups walked according to different patterns. It is suggested that the copers stabilized their knee joint by co-contraction of the hamstrings and quadriceps muscles, while the non-copers lacked this ability. Instead, the non-copers reduced the knee extensor moment in order to decrease anterior displacement of the tibia. The walking pattern differences observed between the copers and non-copers may explain their different post-injury activity levels. PMID- 12736839 TI - The effect of training in male prepubertal and pubertal monozygotic twins. AB - Nine male pairs of monozygotic twins aged 11-14 years, height 147 (7.6) cm and body mass 39.7 (9.6) kg, participated in this study. Twin zygocity was tested using morphological, dermatoglyphic and hematologic methods, and Tanner's five stages were used for the evaluation of biological maturation. One twin from each pair undertook training for 6 months, three times a week, with running at 85-120% of the lactate anaerobic threshold (LT). Anthropometrics, determination of maximum O(2) uptake (.VO(2max)), LT and maximal blood lactate concentration ([La](max)) was carried out before, during and after training. No significant difference existed between the trained twins and their untrained brothers before training. After training, the trained twins increased their .VO(2max), (per kg body mass) by 10.6% and their LT by 18.2% (P<0.01), reaching values that differed significantly from those of their untrained brothers [57.5 (3.6) ml.kg(-1).min( 1) vs 55.4 (3.3) ml.kg(-1).min(-1) and 13.4 (1.1) km.h(-1) vs 12.7 (1.1) km.h( 1), respectively]. In addition, in the trained twins relative body fat was reduced (P<0.05) from 17.8 to 16.2% and their somatotype altered significantly (decrease of endomorphy and mesomorphy and increase of ectomorphy), while in the untrained twins there was no change in these parameters. Both groups of twins significantly increased their absolute .VO(2max) after the 6 months of training, the trained by 14,9% [from 2.08 (0.43) to 2.37 (0.45) l.min(-1)] and the untrained by 10.5% [from 2.10 (0.41) to 2.32 (0.47) l.min(-1)], but no difference was registered between them. A comparison of the intrapair changes in .VO(2max) of prepubertal and pubertal twins showed an influence of training in the prepubertal (19.3% vs 5.2%) but not in the pubertal twins (12.7% vs 13.1%). Using analysis of variance, the relative importance of training, heredity and their interaction was evaluated to be 20%, 70% and 10%, respectively, for the change in body fat, 35%, 45% and 20%, respectively, for the change in relative .VO(2max) and 25-30%, 50-60% and 15-20%, respectively, for the change in LT. In conclusion, training during pubertal growth can favour aerobic power (depending on body composition) as well as aerobic capacity, but it has no effect on absolute .VO(2max). Genetic control seems to have a strong effect on the extent of adaptations, and the genotype-training interaction explains a small, but prominent part of them. PMID- 12736840 TI - Experimental evaluation of eye-blink parameters as a drowsiness measure. AB - Drowsiness and increased tendency to fall asleep during daytime is still a generally underestimated problem. An increased tendency to fall asleep limits the efficiency at work and substantially increases the risk of accidents. Reduced alertness is difficult to assess, particularly under real life settings. Most of the available measuring procedures are laboratory-oriented and their applicability under field conditions is limited; their validity and sensitivity are often a matter of controversy. The spontaneous eye blink is considered to be a suitable ocular indicator for fatigue diagnostics. To evaluate eye blink parameters as a drowsiness indicator, a contact-free method for the measurement of spontaneous eye blinks was developed. An infrared sensor clipped to an eyeglass frame records eyelid movements continuously. In a series of sessions with 60 healthy adult participants, the validity of spontaneous blink parameters was investigated. The subjective state was determined by means of questionnaires immediately before the recording of eye blinks. The results show that several parameters of the spontaneous eye blink can be used as indicators in fatigue diagnostics. The parameters blink duration and reopening time in particular change reliably with increasing drowsiness. Furthermore, the proportion of long closure duration blinks proves to be an informative parameter. The results demonstrate that the measurement of eye blink parameters provides reliable information about drowsiness/sleepiness, which may also be applied to the continuous monitoring of the tendency to fall asleep. PMID- 12736842 TI - Non-invasive evaluation of gas exchange during a shuttle walking test vs. a 6-min walking test to assess exercise tolerance in COPD patients. AB - Walking tests, such as the "shuttle" incremental walking test (SWT) and the 6-min walking test (6'WT), are commonly utilized in evaluating exercise intolerance in patients with chronic obstructive pulmonary disease (COPD) and the distance covered is the variable usually considered. Because lung gas exchange indexes are not measured, little is known about the physiological response elicited by different walking protocols. We compared exercise adaptation during the 6'WT and SWT in 13 male stable COPD patients [mean (SE) age: 70 (1) years; forced expiratory volume in 1 s (FEV(1)): 1.2 (0.1) l; arterial O(2) tension (PaO(2)): 72 (2) mmHg; arterial CO(2) tension (PaCO(2)): 41 (1) mmHg]. Oxygen uptake (.VO(2)), CO(2) output (.VCO(2)), minute ventilation (.V(E)), and heart rate (HR) were monitored by a portable telemetric system. During the SWT a linear response in lung gas exchange indexes was observed while, during the 6'WT, the response was exponential. During the 6'WT, .VO(2), .VCO(2), .V(E), and HR values at steady state (SS) were significantly lower compared to SWT peak values. For SWT, distance covered correlated with .VO(2PEAK), (R=0.86, p<0.001), .VCO(2PEAK), (R=0.87, p<0.001) and .V(EPEAK) (R=0.74, p<0.01); moreover, distance and .VO(2PEAK) were significantly correlated with peak .VO(2) values obtained during cycle ergometer incremental exercise (R=0.72, p<0.01 and R=0.92, p<0.0001, respectively). For 6'WT, the distance covered did not correlate with any pertinent physiological index. The two walking protocols reveal substantial differences in pathophysiologic adaptations and provide evidence that SWT is more accurate than the 6'WT in the evaluation of maximal exercise tolerance in COPD patients. PMID- 12736841 TI - Changes in the somatosensory N250 and P300 by the variation of reaction time. AB - We investigated the relationship between somatosensory event-related potentials (ERP) and the variation of reaction time (RT). For this purpose, we recorded the ERPs (N250 and P300) in fast- and slow-reaction trials during a somatosensory discrimination task. Strong, standard, and weak target electrical stimuli were randomly delivered to the left median nerve at the wrist with a random interstimulus interval (900-1,100 ms). All the subjects were instructed to respond by pressing a button with their right thumb as fast as possible whenever a target stimulus was presented. We divided all the trials into fast- and slow-RT trials and averaged the data. N250 latency tended to be delayed when the RT was slow, but not significantly. P300 latency was delayed significantly when the RT was slow, but to a much lesser extent than the RT delay, so we concluded that the change of RT was not fully determined by the processes reflected by the somatosensory N250 or P300. Furthermore, the larger and earlier P300 in the fast RT trials implied that when larger amounts of attentional resources were allocated to a given task, the speed of stimulus evaluation somewhat increased and RT was shortened to a great extent. N250 amplitude did not significantly vary in the two RT clusters. In conclusion, the somatosensory N250 reflects active target detection, which is relatively independent of the modulation of the response speed, whereas the somatosensory P300 could change without manipulation of either the stimulus or the response processing demand. PMID- 12736843 TI - Optimising high-intensity treadmill training using the running speed at maximal O(2) uptake and the time for which this can be maintained. AB - The aim of this study was to compare the effects of two high-intensity, treadmill interval-training programs on 3000-m and 5000-m running performance. Maximal oxygen uptake (.VO(2max)), the running speed associated with .VO(2max) (v.VO(2max)), the time for which v.VO(2max) can be maintained (T(max)), running economy (RE), ventilatory threshold (VT) and 3000-m and 5000-m running times were determined in 27 well-trained runners. Subjects were then randomly assigned to three groups; (1) 60% T(max), (2) 70% T(max) and (3) control. Subjects in the control group continued their normal training and subjects in the two T(max) groups undertook a 4-week treadmill interval-training program with the intensity set at v.VO(2max) and the interval duration at the assigned T(max). These subjects completed two interval-training sessions per week (60% T(max)=six intervals/session, 70% T(max) group=five intervals/session). Subjects were re tested on all parameters at the completion of the training program. There was a significant improvement between pre- and post-training values in 3000-m time trial (TT) performance in the 60% T(max) group compared to the 70% T(max) and control groups [mean (SE); 60% T(max)=17.6 (3.5) s, 70% T(max) =6.3 (4.2) s, control=0.5 (7.7) s]. There was no significant effect of the training program on 5000-m TT performance [60% T(max)=25.8 (13.8) s, 70% T(max)=3.7 (11.6) s, control=9.9 (13.1) s]. Although there were no significant improvements in .VO(2max), v.VO(2max) and RE between groups, changes in .VO(2max) and RE were significantly correlated with the improvement in the 3000-m TT. Furthermore, VT and T(max) were significantly higher in the 60% T(max) group post- compared to pre-training. In conclusion, 3000-m running performance can be significantly improved in a group of well-trained runners, using a 4-week treadmill interval training program at v.VO(2max) with interval durations of 60% T(max). PMID- 12736844 TI - Impact of elevated ambient temperatures on the acute immune response to intensive endurance exercise. AB - To date, there has been little research examining how elevated ambient temperatures exert an additional effect on the acute immune response to endurance exercise. Seven endurance-trained, non-heat-acclimated men [mean (95% confidence interval): 29.7 (25.9-33.5) years, .VO(2max) 66.3 (61.3-71.3) ml.kg(-1).min(-1)] performed two 60-min treadmill runs (75% .VO(2max)) in two different environments (EX1: 18 degrees C/50% room temperature/relative humidity and EX2: 28 degrees C/50% room temperature/relative humidity) with a 7-day interval between the runs. Blood samples were drawn at rest and 0, 0.5, 3, 24, and 48 h after exercise. Compared to EX1, exercise-induced increases in core temperature, sweating rate, heart rate, plasma norepinephrine, cortisol, human growth hormone, and neutrophil and monocyte counts were significantly (5% level) more pronounced after EX2. In contrast, responses of plasma epinephrine, myeloperoxidase, interleukin (IL)-6 as well as lymphocyte counts were similar in EX1 and EX2. Plasma concentrations of IL-8 and C-reactive protein were affected by neither exercise nor by additional heat exposure. Our results suggest that the additional impact of elevated ambient temperatures on stress responses to endurance exercise in trained subjects seems to affect primarily the cardiocirculatory and hormonal systems, and resulting changes in neutrophil and monocyte cell-trafficking. In contrast, heat stress does not seem to exert large additional effects on the acute immune response to endurance exercise as performed in the present study. PMID- 12736845 TI - Muscle activity during computer-based office work in relation to self-reported job demands and gender. AB - The aim was to investigate whether quantitative job demands influence muscle activity among women, and whether there are gender differences in duration of computer, mouse, and keyboard use and muscle activity of shoulder and forearm muscles during work. The study was carried out in an occupational setting, and 24 women and 11 men from a municipal administration participated. The duration of computer, mouse, and keyboard use was measured by a commercial software package. Quantitative job demands were registered by questionnaire. Electromyography (EMG) was measured bilaterally from the upper trapezius and the extensor digitorum communis muscles. No association was found between self-reported quantitative job demands and muscle activity among the women ( n=24). The women used the keyboard more frequently ( p=0.020) and tended to perform fewer mouse clicks compared to men ( p=0.057), but no difference was seen in EMG activity between men ( n=11) and women ( n=11) from the same department. However, office assistants (six women) showed significantly higher static EMG activity levels ( p=0.042) and almost significantly shorter EMG gap times ( p=0.060) than the rest of the subjects (5 women and 11 men). This indicated that shorter muscular resting periods among female office assistants as compared to the other subjects were due to differences in job content rather than gender differences. PMID- 12736846 TI - Effects of one night's sleep deprivation on anaerobic performance the following day. AB - The purpose of this study was to determine the effect of one night's sleep deprivation on anaerobic performance in the morning and afternoon of the following day. Thirteen healthy males were studied twice in a balanced, randomized design. The experiment consisted of two conditions 1 week apart. In the sleep deprivation condition (SDN) subjects remained awake overnight and in the control condition (reference night, RN) the same subjects slept at home, retiring between 2230 and 2330 hours, as decided individually, and rising at 0500 hours. In both conditions, activity, sleep and diet were monitored by actimetry and daily activity and dietary diaries. Physical performance testing was carried out at 0600 hours and at 1800 hours after the one night of sleep and the one night of sleep deprivation. At each test occasion, subjects were measured for maximal power ( P(max)), peak power ( P(peak)) and mean power ( P(mean)). Blood lactate concentrations were measured at rest, at the end of the force-velocity ( F- V) test, just before and just after the Wingate test and again 5 min later. Oral temperatures were measured every 2 h. In both conditions, the results showed a circadian rhythm in temperature. Analysis of variance revealed a significant (sleep x time of day of test) interaction effect on P(peak), P(mean) and P(max). These variables improved significantly from morning to afternoon after RN and SDN. The reference night was followed by a greater improvement than the SDN. Up to 24 h of waking, anaerobic power variables were not affected; however, they were impaired after 36 h without sleep. Analysis of variance revealed that blood lactate concentrations were unaffected by sleep loss, by time of day of testing or by the interaction of the two. In conclusion, sleep deprivation reduced the difference between morning and afternoon in anaerobic power variables. Anaerobic performances were unaffected after 24 h of wakefulness but were impaired after 36 h without sleep. PMID- 12736847 TI - EMG power spectra of cervical muscles in lateral flexion and comparison with sagittal and oblique plane activities. AB - The purpose of this study was to calculate electromyographic (EMG) power spectra of the sternocleidomastoid (SCM), the splenius capitis (SPL) and the trapezius (TRP) muscles from both sides of the body in bilateral lateral flexion and compare them with the EMG spectral characteristics of the same muscles in the sagittal and oblique planes recorded from the same subjects in the same experimental session but reported elsewhere in the literature. Forty normal and healthy young adults (21 males, 19 females) volunteered for the study. The subjects were prepared and positioned appropriately to exert linearly ramping isometric exertion aided by visual feedback against a rigid and load cell instrumental device. Exertions were carried out in a random order. The maximal voluntary contraction was reached within a 5-s test period. The torque and EMG from the SCM, SPL and TRP muscles were sampled bilaterally at a rate of 1 kHz. The EMG data were subjected to fast Fourier transform analysis. During lateral flexion, the torque generated by females was significantly lower than males ( P<0.01) and represented at 75% of male torque. Contrary to flexion/extension and oblique plane activities, the ipsilateral SCM was most active with highest power and up to four dominant frequencies demonstrating four distinct peaks. The bandwidth of this muscle progressively increased with the grade of contraction up to 400 Hz, similar to flexion/extension with median frequency (MF) spread between 21 and 109 Hz for males and 78 and 99 Hz for females. The ipsilateral SPL was the second most active muscle, which maintained a narrow bandwidth (200 Hz) and one dominant frequency, rising in power with increasing grade of contraction. Flexion and extension caused symmetrical activity bilaterally. The results of MANOVA revealed a significant main effect of gender, activity direction, grade of contraction and individual muscles ( P<0.001). There was a significant interaction between muscle and activity direction ( P<0.001) implying significantly different MF and mean power frequency if the direction of exertion was changed. Male and female subjects responded to direction differently ( P<0.001). PMID- 12736848 TI - Skin-temperature gradients are a validated measure of fingertip perfusion. PMID- 12736852 TI - [Infectious disease as a specialty--a Swiss perspective]. PMID- 12736853 TI - [Nocardia cyriacigeorgici: First report of invasive human infection]. AB - BACKGROUND: Diagnostic laboratories increasingly offer bacterial identification to the species level. The 17 nocardia species known to date differ in their clinical presentation, antibiotic resistance patterns and geographic distribution. The discovery of a new species with pathogenicity for humans calls for the characterization of its clinical and epidemiological properties. PATIENTS AND METHODS: Nocardia isolated from multifocal brain abscesses of an immunocompromised patient were further identified by the analysis of their cellular fatty acids and sequencing of the 16S ribosomal DNA. Quantitative antibiotic resistance testing was performed with E-tests. RESULTS: The 16S ribosomal DNA analysis showed a 99 % homology to Nocardia cyriacigeorgici. This is the first report of this species as an invasive human pathogen. N. cyriacigeorgici was found susceptible for meropenem, amikacin, ceftriaxon and cotrimoxazole. The combination of surgical drainage and antibiotic treatment for 13 months was curative. CONCLUSIONS: N. cyriacigeorgici has the potential to cause invasive infections at least in immunocompromised patients. Comparing clinical and in vitro characteristics with N. asteroides, the main causative agent of nocardial infections in Europe, we found no clinically relevant differences. PMID- 12736854 TI - [Fever after a tick bite: clinical manifestations and diagnosis of acute tick bite-associated infections in northeastern Switzerland]. AB - BACKGROUND: Different tick-borne infections can cause an acute febrile illness. The study objectives were to investigate the clinical manifestations and diagnosis of infections among patients who presented with fever after a tick bite, and to detect newly described pathogens, including Ehrlichia, Babesia and Rickettsia helvetica, in North-Eastern Switzerland. PATIENTS AND METHODS: : We studied 75 patients (41 male, 34 female, median age 38 years, among them 10 children) who had fever within 3 weeks after a tick-bite. Paired sera were tested for antibodies to Borrelia burgdorferi, tick-borne encephalitis virus, Anaplasma (Ehrlichia) phagocytophila, Babesia microti, B. divergens, and Rickettsia helvetica. In addition, microscopy and polymerase chain reaction was used to detect Ehrlichia. Clinical data were obtained at baseline and at 1 and 2 year follow-up. RESULTS: Tick-borne infections were confirmed or possible in 36 (48 %) patients: 7 (9 %) Erythema migrans, 6 (8 %) other specific manifestations of Lyme borreliosis, 6 (8 %) Lyme borreliosis presenting as non-specific febrile illness, 8 (11 %) tick-borne encephalitis, 7 (10 %) granulocytic ehrlichiosis, 1 B. microti infection in a traveler from the US and 6 (8 %) dual infections. In 8 (11 %) patients serological findings were suggesting possible acute or past R. helvetica infection. CONCLUSION: Among patients with fever after a tick-bite, Lyme borreliosis was most frequently found. There was no evidence for babesiosis among the resident population. Serologic data suggest that human granulocytic ehrlichiosis and R. helvetica infections may be endemic in Switzerland. Among 50 % of the patients no tick-borne infections could be diagnosed. PMID- 12736855 TI - [Candida albicans endocarditis. A rare disease with serious complications]. AB - HISTORY AND ADMISSION FINDINGS: A 54-year-old farmer with moderate mitral valve regurgitation was admitted to hospital because of suspected infective endocarditis. EXAMINATIONS: Echocardiography revealed a large mitral valve vegetation as the source of multifocal septic emboli to the central nervous system, spleen, mesenteric and femoro-popliteal arteries, eyes, and kidneys. Eventually an embolus removed from the femoro-popliteal artery and vegetations on the replaced mitral valve grew C. albicans. THERAPY: Despite treatment with amphotericin B and valve replacement the patient died of septicemia due to E. coli. CONCLUSION: Endocarditis due to C. albicans is commonly associated with severe complications. Diagnosis of this rare disease is often delayed because of negative blood cultures. Large cardiac vegetations and embolization of major arterial vessels should raise the suspicion of fungal endocarditis. PMID- 12736856 TI - [HIV-infection, HAART (highly-active antiretroviral therapy) and hyperlipidemia]. AB - HISTORY AND ADMISSION FINDINGS: A 42-year-old man with long-standing HIV infection (CDC C3) and multiple treatment failures was seen for follow-up 6 months after starting a new HAART (zidovudine, efavirenz, indinavir, ritonavir). The medical history is remarkable for a hypersensitivity reaction to abacavir, past smoking (10 pack years) and a family history of coronary heart disease of the patient's mother. Physical examination revealed discrete signs of lipoatrophy of the face, arms and legs, but no lipoaccumulation (hip-waist ratio 0.94; body mass index 22 kg/m2). Blood pressure measured 156/92 mmHg. LABORATORY FINDINGS: A rise in CD4 cell count from 24/ microL to 60/ microL was noted. For the first time, the HIV viral load decreased to < 50 copies/mL. Concentrations of triglycerides and cholesterol were found to be increased to 26 mmol/L (2275 mg/dL) and 15 mmol/L (580 mg/dL), respectively, compared previously normal readings. Other parameters were in the normal range. DIAGNOSIS AND THERAPY: Continuation of HAART resulted in rising CD4 counts to > 200/ microL and HIV viral load suppression to < 50 copies/mL. Neither dietary measures nor treatment with pravastatin significantly changed the drug-induced mixed hyperlipidemia. Treatment with fenofibrate (200 mg qd), however, was followed by a significant reduction of triglyceride and cholesterol concentrations to 12 mmol/L (1050 mg/dL) and 10 mmol/L (387 mg/dL). Addition of pravastatin did not result in further improvement. Arterial hypertension was diagnosed by ambulatory 24-h blood pressure monitoring (systolic 157+/-11 mmHg; diastolic 97+/-10 mmHg). CONCLUSION: HAART-induced mixed hyperlipidaemia is likely to increase the cardiovascular risk. According to first estimates, the 3-year risk of myocardial infarction amounts to 1.29 - 2.09 % for our patient. The 3-year risk of HIV progression to AIDS or death is > 30 % if HAART is stopped or fails upon switching due to unknown archived drug resistance. Thus, despite unsatisfactory lipid profiles, a modification of HAART seems only acceptable, if new, equally potent, but less lipidaemic drug combinations are available. PMID- 12736857 TI - [Diagnosis and treatment of bites by cats, dogs and humans]. PMID- 12736858 TI - [Challenges in HIV long-term care]. PMID- 12736859 TI - [New quinolones for the treatment of community-acquired pneumonia--pro]. PMID- 12736861 TI - [How can factor X deficiency in nephropathy explained?]. PMID- 12736860 TI - [New quinolones for the treatment of community-acquired pneumonia--contra]. PMID- 12736862 TI - [German-Austrian guidelines for diagnosis and treatment]. PMID- 12736863 TI - [German-Austrian guidelines for antiretroviral therapy of HIV infection. (July, 2002, update)]. PMID- 12736864 TI - [German-Austrian guidelines for HIV therapy during pregnancy (May, 2001, update]. PMID- 12736865 TI - [German-Austrian guidelines for diagnosis and treatment of HIV-discordant couples who wish to have children]. PMID- 12736866 TI - [German-Austrian guidelines for postexposure prophylaxis of HIV infection. (May, 2002, update)]. PMID- 12736867 TI - Sequence analysis of the mitochondrial genomes from Dutch pedigrees with Leber hereditary optic neuropathy. AB - The complete mitochondrial DNA (mtDNA) sequences for 63 Dutch pedigrees with Leber hereditary optic neuropathy (LHON) were determined, 56 of which carried one of the classic LHON mutations at nucleotide (nt) 3460, 11778, or 14484. Analysis of these sequences indicated that there were several instances in which the mtDNAs were either identical or related by descent. The most striking example was a haplogroup J mtDNA that carried the 14484 LHON mutation. Four different but related mitochondrial genotypes were identified in seven of the Dutch pedigrees with LHON, including six of those described by van Senus. The control region of the founder sequence for these Dutch pedigrees with LHON matches the control region sequence that Macmillan and colleagues identified in the founder mtDNA of French Canadian pedigrees with LHON. In addition, we obtained a perfect match between the Dutch 14484 founder sequence and the complete mtDNA sequences of two Canadian pedigrees with LHON. Those results indicate that these Dutch and French Canadian 14484 pedigrees with LHON share a common ancestor, that the single origin of the 14484 mutation in this megalineage occurred before the year 1600, and that there is a 14484/haplogroup J founder effect. We estimate that this lineage--including the 14484 LHON mutation--arose 900-1,800 years ago. Overall, the phylogenetic analyses of these mtDNA sequences conservatively indicate that a LHON mutation has arisen at least 42 times in the Dutch population. Finally, analysis of the mtDNA sequences from those pedigrees that did not carry classic LHON mutations suggested candidate pathogenic mutations at nts 9804, 13051, and 14325. PMID- 12736868 TI - Multiple mutations of MYO1A, a cochlear-expressed gene, in sensorineural hearing loss. AB - Myosin I isozymes have been implicated in various motile processes, including organelle translocation, ion-channel gating, and cytoskeleton reorganization. Unconventional myosins were among the first family of proteins found to be associated with hearing loss in both humans and mice. Here, we report the identification of a nonsense mutation, of a trinucleotide insertion leading to an addition of an amino acid, and of six missense mutations in MYO1A cDNA sequence in a group of hearing-impaired patients from Italy. MYO1A, which is located within the DFNA48 locus, is the first myosin I family member found to be involved in causing deafness and may be a major contributor to autosomal dominant-hearing loss. PMID- 12736870 TI - Disruption of the serine/threonine kinase 9 gene causes severe X-linked infantile spasms and mental retardation. AB - X-linked West syndrome, also called "X-linked infantile spasms" (ISSX), is characterized by early-onset generalized seizures, hypsarrhythmia, and mental retardation. Recently, we have shown that the majority of the X-linked families with infantile spasms carry mutations in the aristaless-related homeobox gene (ARX), which maps to the Xp21.3-p22.1 interval, and that the clinical picture in these patients can vary from mild mental retardation to severe ISSX with additional neurological abnormalities. Here, we report a study of two severely affected female patients with apparently de novo balanced X;autosome translocations, both disrupting the serine-threonine kinase 9 (STK9) gene, which maps distal to ARX in the Xp22.3 region. We show that STK9 is subject to X inactivation in normal female somatic cells and is functionally absent in the two patients, because of preferential inactivation of the normal X. Disruption of the same gene in two unrelated patients who have identical phenotypes (consisting of early-onset severe infantile spasms, profound global developmental arrest, hypsarrhythmia, and severe mental retardation) strongly suggests that lack of functional STK9 protein causes severe ISSX and that STK9 is a second X chromosomal locus for this disorder. PMID- 12736872 TI - Complications of fluoroscopically guided interlaminar cervical epidural injections. AB - OBJECTIVES: To assess the incidence of complications of fluoroscopically guided interlaminar cervical epidural injections. DESIGN: A retrospective cohort design study. SETTING: A multidisciplinary spine care center. PARTICIPANTS: One hundred fifty-seven consecutive patients with cervical radicular pain caused by cervical spondylosis or herniated nucleus pulposus confirmed by magnetic resonance imaging or computed tomography scanning. INTERVENTIONS: Fluoroscopically guided interlaminar cervical epidural injections were performed at the C7-T1 or C6-7 level using an 18-gauge, 9-mm Tuohy needle with 2mL of 1% lidocaine (Xylocaine) and 80-mg of triamcinolone acetonide (Kenalog). All injections were performed consecutively over a 12-month period by 1 of 5 physicians. MAIN OUTCOME MEASURES: An independent observer reviewed medical charts, which included a 24-hour postprocedure telephone call by an ambulatory surgery center nurse who asked a standardized questionnaire about complications after the injections. Also reviewed were physician notes regarding office follow-up consultations 3 weeks or less after the injections and epidurograms. RESULTS: The charts of 157 patients, who received 345 injections, were reviewed. Complications per injection included 23 increased neck pain (6.7%), 16 transient nonpositional headaches that resolved within 24 hours (4.6%), 6 episodes of insomnia the night of the injection (1.7%), 6 vasovagal reactions (1.7%), 5 facial flushing (1.5%), 1 fever the night of the procedure (0.3%), and 1 dural puncture (0.3%). The incidence of all complications per injection was 16.8%. CONCLUSIONS: Because all complications resolved without morbidity and no patient required hospitalization, fluoroscopically guided interlaminar cervical epidural injections may be a safe procedure for use in patients with cervical radicular pain. PMID- 12736873 TI - Management of knee osteoarthritis: knee lavage combined with hylan versus hylan alone. AB - OBJECTIVE: To assess the difference in efficacy between knee lavage plus the standard hylan G-F 20 (a derivative of hyaluronan) protocol and the standard hylan G-F 20 as per standard usage protocol alone for the treatment of knee osteoarthritis (OA). DESIGN: Nonrandomized prospective study in which patients chose their treatment group. Follow-up averaged 1.1 years. SETTING: Faculty practice of a single physician at a major teaching hospital. PARTICIPANTS: Eighty one patients with documented knee OA on magnetic resonance imaging. INTERVENTIONS: Group 1 (n=44) received a single-needle lavage 1 week before the standard hylan G-F 20 protocol; group 2 (n=37) received the standard hylan G-F 20 protocol alone. MAIN OUTCOME MEASURES: Pre- and posttreatment scores on the Lysholm-II Questionnaire and a visual analog scale (VAS) were documented for each patient. The Wilcoxon signed-rank test was used for statistical analysis. RESULTS: A successful outcome was noted in 79.5% of group 1 patients and in 54% of group 2 patients (P<.05). CONCLUSIONS: In the management of knee OA, the use of knee lavage before viscosupplementation with hylan G-F 20 yields better results than using hylan G-F 20 alone. The presence of radiologic grade IV knee OA or moderate to severe patellofemoral arthritis are negative prognostic factors. PMID- 12736871 TI - Genomewide scan for hand osteoarthritis: a novel mutation in matrilin-3. AB - Osteoarthritis (OA) is the most common human joint disease, characterized by loss and/or remodeling of joint synovium, cartilage, and bone. Here, we describe a genomewide linkage analysis of patients with idiopathic hand OA who were carefully phenotyped for involvement of either or both the distal interphalangeal (DIP) joints and the first carpometacarpal (CMC1) joints. The best linkage peaks were on chromosomes 4q and 3p and on the short arm of chromosome 2. Genomewide significance was reached for a locus on chromosome 2 for patients with affected CMC1 joints (LOD = 4.97); this locus was also significant for patients with OA in both CMC1 and DIP joints (LOD = 4.44). The peak LOD score at this locus coincides with a gene, MATN3, encoding the noncollagenous cartilage extracellular matrix protein, matrilin-3. Subsequent screening of the genomic sequence revealed a missense mutation, of a conserved amino acid codon, changing threonine to methionine in the epidermal growth factor-like domain in matrilin-3. The missense mutation cosegregates with hand OA in several families. The mutation frequency is slightly more than 2% in patients with hand OA in the Icelandic population and has a relative risk of 2.1. PMID- 12736874 TI - Cyproheptadine for intrathecal baclofen withdrawal. AB - OBJECTIVE: To evaluate the efficacy of cyproheptadine in the management of acute intrathecal baclofen (ITB) withdrawal. DESIGN: Descriptive case series. SETTING: University hospital with a comprehensive in- and outpatient rehabilitation center. PARTICIPANTS: Four patients (3 with spinal cord injury, 1 with cerebral palsy) with implanted ITB infusion pumps for treatment of severe spasticity, who had ITB withdrawal syndrome because of interruption of ITB infusion. INTERVENTIONS: Patients were treated with 4 to 8mg of cyproheptadine by mouth every 6 to 8 hours, 5 to 10mg of diazepam by mouth every 6 to 12 hours, 10 to 20mg of baclofen by mouth every 6 hours, and ITB boluses in some cases. MAIN OUTCOME MEASURES: Clinical signs and symptoms of ITB withdrawal of varying severity were assessed by vital signs (temperature, heart rate), physical examination (reflexes, tone, clonus), and patient report of symptoms (itching, nausea, headache, malaise). RESULTS: The patients in our series improved significantly when the serotonin antagonist cyproheptadine was added to their regimens. Fever dropped at least 1.5 degrees C, and heart rate dropped from rates of 120 to 140 to less than 100bpm. Reflexes, tone, and myoclonus also decreased. Patients reported dramatic reduction in itching after cyproheptadine. These changes were associated temporally with cyproheptadine dosing. DISCUSSION: Acute ITB withdrawal syndrome occurs frequently in cases of malfunctioning intrathecal infusion pumps or catheters. The syndrome commonly presents with pruritus and increased muscle tone. It can progress rapidly to high fever, altered mental status, seizures, profound muscle rigidity, rhabdomyolysis, brain injury, and death. Current therapy with oral baclofen and benzodiazepines is useful but has variable success, particularly in severe cases. We note that ITB withdrawal is similar to serotonergic syndromes, such as in overdoses of selective serotonin reuptake inhibitors or the popular drug of abuse 3,4 methylenedioxymethamphetamine (Ecstasy). We postulate that ITB withdrawal may be a form of serotonergic syndrome that occurs from loss of gamma-aminobutyric acid B receptor-mediated presynaptic inhibition of serotonin. CONCLUSION: Cyproheptadine may be a useful adjunct to baclofen and benzodiazepines in the management of acute ITB withdrawal syndrome. PMID- 12736875 TI - Intrathecal baclofen in subjects with spastic hemiplegia: assessment of the antispastic effect during gait. AB - OBJECTIVE: To determine whether leg muscle stiffness is measurably reduced after intrathecal baclofen (ITB) in subjects with spastic hemiplegia. DESIGN: Nonrandomized trial. SETTING: Inpatient multidisciplinary rehabilitation unit in France. PARTICIPANTS: Seven consecutive subjects with spastic hemiplegia having Ashworth Scale scores for their quadriceps and triceps greater than 2. INTERVENTION: Subjects were given ITB by lumbar puncture after a dose-selecting test period. MAIN OUTCOME MEASURES: Triceps and quadriceps Ashworth scores, gait analysis at preferred and maximal speed measured by a motion analysis system with 2 forceplates, and electromyographic recording of leg muscles before and 4 hours after ITB. The slopes of the moment-angle curves were measured on the hemiplegic side at the onset of ankle and knee flexion to assess muscle stiffness during walking. Pre- and post-ITB spatiotemporal, kinetic, and kinematic data were compared by using a nonparametric test (Wilcoxon signed-rank test). RESULTS: Ashworth scores of the quadriceps and triceps of all subjects decreased significantly after ITB. Maximal walking speed increased significantly, with a significant increase in stride length, but the preferred walking speed was unchanged. Minimal knee extension and maximal ankle flexion were the only kinematic data significantly different (increased) after ITB. The slope of the ankle moment-angle curve decreased significantly after ITB at preferred gait speed; it also decreased at maximal gait speed in all but 1 subject. Of the 4 available moment-angle curves, 3 showed decreased knee extensor muscle stiffness. The duration of the bursts of spastic muscles decreased after ITB. CONCLUSION: Acute ITB improved walking and reduced muscle stiffness at both the ankles and knees on the spastic hemiplegic side of our subjects. Electromyographic findings suggest that some of the post-ITB reduction in muscle stiffness might be attributed to decreased spasticity. PMID- 12736876 TI - Influence of nonsteroidal anti-inflammatory drug treatment duration and time of onset on recovery from exercise-induced muscle damage in rats. AB - OBJECTIVE: To determine if duration and time of onset of treatment with diclofenac sodium influence force recovery after exercise-induced muscle damage in rats. DESIGN: Randomized placebo-controlled trial. SETTING: Animal laboratory. ANIMALS: A total of 217 female adult Wistar rats. INTERVENTION: Rats were submitted to a protocol consisting of 450 eccentric contractions of the ankle dorsiflexors. Treatment by gavage with diclofenac sodium (1 mg/kg, twice daily) was started at different times pre- and postprotocol or for various treatment durations. MAIN OUTCOME MEASURES: In vitro contractile properties. RESULTS: When treatment was initiated shortly postprotocol, force recovery was roughly proportional to treatment duration during the first 3 days but not at 7 and 28 days postprotocol. A 7-day treatment was no more effective than 1- or 2-day treatments when force was measured at 7 and 28 days; however, such prolonged treatment had no deleterious effect on muscle force at either time. A single-dose prophylactic treatment was as effective as a 2-day treatment initiated soon after the protocol when force was assessed 2 days postprotocol; on the other end, a treatment delayed for 3 days had no effect when force was measured at 7 days. CONCLUSIONS: Treatment with diclofenac sodium extending past the acute inflammatory phase was no more effective than short and timely treatment in this model of skeletal muscle damage. PMID- 12736877 TI - Psychometric properties of the Activities-specific Balance Confidence Scale among individuals with a lower-limb amputation. AB - OBJECTIVE: To evaluate the internal consistency, test-retest reliability, and construct validity of the Activities-specific Balance Confidence (ABC) Scale among people who have a lower-limb amputation. DESIGN: Retest design. SETTING: A university-affiliated outpatient amputee clinic in Ontario. PARTICIPANTS: Two samples of individuals who have unilateral transtibial and transfemoral amputation. Sample 1 (n=54) was a consecutive and sample 2 (n=329) a convenience sample of all members of the clinic population. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Repeated application of the ABC Scale, a 16-item questionnaire that assesses confidence in performing various mobility-related tasks. Correlation to test hypothesized relationships between the ABC Scale and the 2-minute walk (2MWT) and the timed up-and-go (TUG) tests; and assessment of the ability of the ABC Scale to discriminate among groups based on amputation cause, amputation level, mobility device use, automatic stepping ability, wearing time, stair climbing ability, and walking distance. RESULTS: Test-retest reliability (intraclass correlation coefficient) of the ABC Scale was .91 (95% confidence interval [CI], .84-.95) with individual item test-retest coefficients ranging from .53 to .87. Internal consistency, measured by Cronbach alpha, was .95. Hypothesized associations with the 2MWT and TUG test were observed with correlations of .72 (95% CI, .56-.84) and -.70 (95% CI, -.82 to -.53), respectively. The ABC Scale discriminated between all groups except those based on amputation level. CONCLUSIONS: Balance confidence, as measured by the ABC Scale, is a construct that provides unique information potentially useful to clinicians who provide amputee rehabilitation. The ABC Scale is reliable, with strong support for validity. Study of the scale's responsiveness is recommended. PMID- 12736878 TI - Mechanical effects of immobilization on the Achilles' tendon. AB - OBJECTIVE: To measure the biomechanical effects of immobilization on the Achilles' tendon. DESIGN: Experimental, controlled study. SETTING: Physiatry research laboratory. ANIMALS: Twenty adult rabbits. INTERVENTION: One hindlimb immobilized in a cast for 4 weeks (n=10) or 8 weeks (n=10). The contralateral legs (n=20) served as controls. MAIN OUTCOME MEASURES: Cross-sectional Achilles' tendon area, mode of failure, mean failure load, and tendon stiffness. RESULTS: The Achilles' tendon cross-sectional area did not change. Achilles' tendon calcaneus units failed at insertion at 4 weeks and in controls. Calcaneus fracture was the most prevalent mode of failure at 8 weeks. The mean load to failure of both immobilized groups (4wk: 187.5+/-45.7N; 8wk: 162.6+/-39.3N) was significantly smaller than that of the control group (549.2+/-93.7N, both P<.005). The mean tendon stiffness of both immobilized groups (4wk: 64.6+/ 24.8N/mm; 8wk: 53.9+/-19.9N/mm) was significantly lower than that of the control group (125.1+/-26.5N/mm, both P<.005). CONCLUSIONS: Immobility for up to 8 weeks caused decreased stiffness but no atrophy or rupture of the Achilles' tendon, suggesting that immobility does not constitute a risk factor for Achilles' tendon midsubstance rupture. Clinically, graded reloading is required after immobilization to restore the tendon insertion and to reverse calcaneal disuse osteoporosis. PMID- 12736879 TI - Reliable serial measurement of cognitive processes in rehabilitation: the Cognitive Log. AB - OBJECTIVE: To evaluate the reliability and utility of a brief quantitative measure of cognitive recovery, the Cognitive Log (Cog-Log), developed for daily use with rehabilitation inpatients to provide information about the recovery of higher neurocognitive processes including verbal recall, attention, working memory, motor sequencing, and response inhibition. DESIGN: Descriptive study of the Cog-Log's normative scores, reliability (interrater, internal consistency), and validity as shown by its relationship to standard neuropsychologic measures. SETTING: Inpatient rehabilitation hospital affiliated with a large university medical center. PARTICIPANTS: One hundred fifty neurorehabilitation inpatients with acquired brain injury; 83 young adults without acquired brain injury were included to provide normative data. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: The Cog-Log; standardized neuropsychologic measures of memory (Wechsler Memory Scale-Revised, Rey Auditory Verbal Learning Test), language, attention (Wechsler Adult Intelligence Scale-Revised), and reasoning (Trail Making Test). RESULTS: Reliability analysis showed strong interrater reliability across items (Spearman r, .749-1.00) and high internal consistency (Cronbach alpha=.778). Factor analysis of the Cog-Log using principal components extraction revealed a unitary factor (eigenvalue=3.48). Cog-Log items designed to measure working memory and immediate and delayed verbal memory were most strongly predictive of performance on similar standardized neuropsychologic measures administered on the same day. CONCLUSION: The Cog-Log appears to be a reliable and efficient tool for measuring ongoing neurocognitive recovery during inpatient rehabilitation. PMID- 12736880 TI - The Erlangen Fitness Osteoporosis Prevention Study: a controlled exercise trial in early postmenopausal women with low bone density-first-year results. AB - OBJECTIVE: To investigate the effect of a 2-year vigorous, combined high-impact, strength, and endurance training program on bone mineral density (BMD) determined by dual-energy x-ray absorptiometry (DXA), quantitative computed tomography (QCT), and ultrasound in early postmenopausal women with osteopenia. DESIGN: Nonrandomized controlled trial, reporting 1-year data. SETTING: Community. PARTICIPANTS: Early postmenopausal (1-8y postmenopausal) osteopenic women (DXA T score at lumbar spine or total hip between -1 and -2.5 standard deviations). The exercise group (n=59; mean age, 55.1+/-3.4y) and control group (n=41; mean age, 55.9+/-3.1y) were recruited from community registers. INTERVENTION: Fourteen months of exercise training, with 2 joint sessions and 2 additional home training sessions. Exercise and control groups were supplemented individually with calcium and cholecalciferol up to 1500mg of calcium and 500IU of vitamin D per day. MAIN OUTCOMES MEASURES: BMD at the lumbar spine and total hip measured by DXA, isometric maximum strength, and maximal oxygen consumption (VO2max) during a stepwise running test to exhaustion. RESULTS: Bone density increased significantly at the lumbar spine for the exercise group (1.3%, P<.001) and decreased in the control group (-1.2%, P<.01). Differences at the total hip ( 0.3%, not significant vs -0.8%, P<.05) and the femoral neck (-0.8%, P<.05 vs 1.8%, P<.001) were nonsignificant. Changes in isometric maximum strength were significant for each region (grip strength, trunk flexors and extensors, hip flexors, leg adductors and abductors, arm flexors and extensors) in the exercise group (11%-39%) compared with nonrelevant changes (-1.1% to 3.9%) in the control group. Between-group differences were significant (P<.01-.001) for all strength parameters. VO2max increased significantly by 11% (P<.001) in the exercise group but decreased in the control group by 4% (P<.05) while showing significant between-group differences. CONCLUSION: High-intensity exercise training can have a positive influence on bone density in early postmenopausal osteopenic women. PMID- 12736881 TI - Mediolateral sway in single-leg stance is the best discriminator of balance performance for Tai-Chi practitioners. AB - OBJECTIVES: To identify a balance measure that can best distinguish Tai-Chi from non-Tai-Chi practitioners and to examine whether longer Tai-Chi practice results in better balance control. DESIGN: Cross-sectional comparative study. SETTING: Community. PARTICIPANTS: Nineteen Tai-Chi practitioners (who practiced Tai Chi for 30-45min at least 3/wk for >1y) and 19 healthy subjects with regular exercise habits (who practiced Tai Chi for 30-45min at least 3/wk for >1y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Scores on 2 clinical tests (functional reach, gait) and 1 laboratory test (postural sway). RESULTS: Tai-Chi practitioners had better clinical test scores for functional reach, gait speed, stride length, and sway parameters during single-leg stance (P<.05). Sway in mediolateral direction during single-leg stance was the balance performance variable that best discriminated the Tai-Chi group from non-Tai-Chi group. More experience practicing Tai Chi was associated with better balance performance. CONCLUSIONS: Tai-Chi practitioners performed better both in clinical and laboratory tests when compared with subjects who did not practice Tai Chi. More Tai-Chi experience was associated with better postural control. PMID- 12736882 TI - Physical training during intrahepatic chemotherapy. AB - OBJECTIVE: To evaluate the role of strength and endurance training for the muscular, cardiac, respiratory, and immune systems and the quality of life (QOL) during intrahepatic chemotherapy (folinic acid, 5-fluorouracil). DESIGN: Single case. SETTING: Teaching hospital in Germany. PARTICIPANT: An elderly athlete with liver metastasis after resection of a carcinoma of the rectum (pT3, N0, M-liver, G2). INTERVENTION: Strength and endurance training during chemotherapy. MAIN OUTCOME MEASURES: During the intervals between training cycles (14d), beginning in postoperative week 6, a strength and endurance training regimen was performed twice weekly for 13 weeks, with an intensity of 40% to 60% of the maximum postoperative individual power and endurance. Before and after chemotherapy, we checked echocardiograms, resting and exercise electrocardiograms, lung function, natural killer (NK) cells, and the Gastrointestinal Quality of Life Index (GIQLI) scores. RESULTS: The increase in strength was between 0% and 144%. The improvement in endurance expressed by reduction of heart rate and lactate concentration was 10% and 21.5%, respectively. Lung function also improved with regard to forced expiratory volume in 1 second (12.9%), forced vital capacity (11.3%), and inspiratory vital capacity (11.4%). The relative count of the NK cells increased to 27.2%. An improvement in the GIQLI was observed from 109 points (pathologic) to 129 points. CONCLUSION: Strength and endurance training was associated with an increase of physical strength and endurance with positive influence on illness-related QOL. Postoperative physical exercise during regional chemotherapy is beneficial. PMID- 12736883 TI - Nondominant hand performance of the Japanese Trail Making Test and its mirror version. AB - OBJECTIVES: To investigate intermanual differences in the Japanese version of the Trail Making Test (JTMT) and to investigate whether a mirror version of the JTMT test sheet (JTMT-M) should be used when the task is performed with the nondominant hand. DESIGN: Intermanual differences were compared among the JTMT with the dominant hand, the JTMT with the nondominant hand, and the JTMT-M with the nondominant hand. SETTING: Rehabilitation center in a university hospital. PARTICIPANTS: A total of 150 persons (age range, 19-38y) who had no history of neurologic or musculoskeletal impairment. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: The time to complete the task. RESULTS: Nondominant hand performance on the JTMT and JTMT-M did not differ significantly from that of the dominant hand on the JTMT. All left hand data except 1 in each task were within the previously reported reference values obtained from the JTMT with the right hand. CONCLUSIONS: The nondominant hand can be used as an alternative in the JTMT and can be expected to perform comparably to the dominant hand. There is no need to use the JTMT-M even when the task is conducted with the nondominant hand. PMID- 12736884 TI - Cardiac risk factors in polio survivors. AB - OBJECTIVE: To assess the prevalence of dyslipidemia and other risk factors for coronary heart disease in a sample of polio survivors with and without postpoliomyelitis syndrome. DESIGN: Retrospective chart review. SETTING: A multidisciplinary outpatient postpolio clinic. PARTICIPANTS: Eighty-eight consecutive symptomatic postpolio patients, 50 women (mean age, 59.0y; range, 36 81y) and 38 men (mean age, 61.2y; range, 44-83y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Presence of risk factors for coronary heart disease: clinical atherosclerotic disease, male age >or=45 years or female age >or=55 years, history of hypertension (blood pressure >or=140/90mmHg or on antihypertensive medication), diabetes mellitus, cigarette smoking, and high density lipoprotein (HDL) less than 35mg/dL. Obesity (body mass index [BMI], >25kg/m(2)) was assessed as an intervention target. Laboratory values included fasting total cholesterol, HDL, low-density lipoprotein, triglycerides, and glucose. RESULTS: Of the total sample, 61.3% had dyslipidemia. Average HDL cholesterol ratio was 4.01 (women, 3.68; men, 4.55). Forty-four patients (50%) had a history of hypertension or had elevated blood pressure. Seven patients (8%) had a history of diabetes or had elevated fasting blood glucose (>110). Eighteen patients (20.4%) were smokers or had a history of smoking; 9 continued to smoke and 9 had quit smoking. Twenty-five patients (28.4%) were overweight (BMI, >25kg/m(2)). Forty-one patients (46.5%) had more than 1 risk factor for coronary heart disease. Nine of the total sample (10.2%) had a history of heart disease ranging from atrial fibrillation to angina. Only 19 patients had a previous diagnosis of dyslipidemia and only 12 were on a lipid-lowering medication. CONCLUSION: Polio patients have a high prevalence of dyslipidemia. The study sample supports the National Cholesterol Education Program's Adult Treatment Panel III statements that hypercholesterolemia is underdiagnosed and undertreated. The postpolio population carries a high prevalence of 2 or more coronary heart disease risk factors. Evaluation and rehabilitation of polio patients should include screening for dyslipidemia and education about elimination of controllable risk factors. PMID- 12736885 TI - Rasch analysis of the Gross Motor Function Measure: validating the assumptions of the Rasch model to create an interval-level measure. AB - OBJECTIVES: To describe the Rasch analysis of the Gross Motor Function Measure (GMFM-88) and to demonstrate how the assumptions of unidimensionality, sample free measurement, and test-free measurement were validated to create an interval level measure. DESIGN: Cross-sectional and longitudinal (12-mo) data from a prospective study of motor development in children with cerebral palsy (CP) were used for the analysis. SETTING: Motor assessments were completed at 18 children's ambulatory rehabilitation centers in Ontario, Canada, by pediatric physical therapists trained in the use of the GMFM-88. PARTICIPANTS: The first 537 of 682 children enrolled into a longitudinal study of motor development in children with CP. Children had a mean age of 6.43+/-2.75 years (range, 11mo-12y) with varying types and severity of CP. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: The GMFM-88. RESULTS: The Rasch analysis, in conjunction with clinical decisions, identified 66 items from the GMFM-88 that formed a unidimensional measure (GMFM 66). Assumptions of sample-free and test-free measurement were confirmed, and a user-friendly scoring program was developed. CONCLUSIONS: The GMFM-66 is an interval-level measure of gross motor function for children with CP; it should improve the scoring, interpretation, and overall clinical and research utility over the original GMFM. PMID- 12736887 TI - Risks of acute hospital transfer and mortality during stroke rehabilitation. AB - OBJECTIVE: To identify demographic, medical, and functional factors associated with transfer of stroke patients to acute hospital services and/or mortality during stroke rehabilitation. DESIGN: Two case-control studies in which logistic regression was used to control for clinical traits associated with differences in likelihood. SETTING: A total of 542 US inpatient and rehabilitation units. PARTICIPANTS: A total of 64,471 patients discharged during 1995. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Transfer to an acute hospital service and death. RESULTS: There were 5847 (9.1%) acute hospital transfers and 320 (0.5%) deaths. Greater disability at admission was associated with higher odds of both acute hospitalization and mortality. Cardiopulmonary arrest, chest pain, gastrointestinal problems, bleeding disorders, hypercoagulable states, and acute renal difficulties increased the relative odds of acute hospitalization from 3.1 (95% confidence interval [CI], 2.3-4.2) to 12.7 (95% CI, 9.2-17.6). The likelihood of mortality for patients 85 years of age or older was more than 2 fold (2.5; 95% CI, 1.7-3.6) that of patients 65 years of age or younger for blacks, it was nearly 2-fold (1.7; 95% CI, 1.3-2.3) compared with whites, after adjusting for clinical differences. CONCLUSION: Higher likelihoods of mortality among older patients versus younger, black patients versus white, and patients with more rather than less disability at admission suggest the need for greater vigilance in monitoring medical status. PMID- 12736886 TI - Activation of natural killer cell function in recreational athletes with paraplegia during a wheelchair half-marathon race. AB - OBJECTIVE: To investigate a part of the immune homeostasis in recreational athletes with spinal cord injury (SCI) during and after a wheelchair half marathon race. DESIGN: Case-control study in an actual race. SETTING: The half marathon division of an international wheelchair marathon race in Japan. PARTICIPANTS: Seven male wheelchair racers with SCI between T7 and L1. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Changes in the number and function of natural killer (NK) cells and the endocrine effects, including plasma catecholamines and cortisol in blood samples obtained the day before the race, immediately after it, and 1 day after the race. RESULTS: Both the percentage and absolute number of peripheral NK cells did not change significantly throughout the experiment. Mean NK cell cytotoxic activity +/- standard deviation increased significantly, from 45.5%+/-7.5% to 56.1%+/-5.1% (P<.01) immediately after the race and remained increased until the next day. Plasma adrenaline levels were increased (P<.05) immediately after the race and recovered 1 day later; the plasma concentration of cortisol did not change throughout the experiment. CONCLUSIONS: The wheelchair half-marathon race induced activation of NK cell function in recreational athletes with SCI between T7 and L1. PMID- 12736888 TI - Ipsilateral deficits of targeted movements after stroke. AB - OBJECTIVE: To test the hypotheses that targeted movements of both the ipsilateral and the contralateral extremities of stroke survivors would be prolonged compared with those from a control group without stroke, and that the ipsilateral deficit would occur in movements toward small, but not large, targets. DESIGN: Descriptive study. SETTING: Motor performance laboratory. PARTICIPANTS: Convenience sample of right-handed individuals including 10 who were more than 6 months poststroke with Fugl-Meyer Motor Assessment scores greater than 75% for the upper (UEs) and lower (LEs) extremities, and a comparison group of 20 age matched adults without stroke. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: The average time required for the stylus, held with the hand or strapped to the foot, to travel from leaving 1 target to contacting the second target (ie, movement time) and the average time the stylus rested on the target (ie, dwell time). RESULTS: Regardless of target size, movement and dwell times for both UEs of the stroke group were prolonged compared with those of the comparison group. Regardless of target size, dwell time for both LEs of the stroke group was prolonged compared with that of the comparison group. CONCLUSIONS: After stroke, the ipsilateral extremities may show subtle deficits in targeted movements. PMID- 12736889 TI - The effect of diabetes combined with stroke on disability, self-rated health, and mortality in older Mexican Americans: results from the Hispanic EPESE. AB - OBJECTIVE: To examine how diabetes in combination with stroke affects functional activities of daily living (ADLs) and instrumental activities of daily living (IADLs), self-rated health, and 5-year mortality in elderly Mexican Americans with or without other comorbid conditions. DESIGN: Longitudinal study. SETTING: Five southwestern states. PARTICIPANTS: A total of 3050 subjects of age 65 years or older, of whom 23% had diabetes and 6% had a stroke. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: ADL and IADL disabilities, self-rated health, and 5-year mortality. RESULTS: Subjects with both diabetes and stroke but without other comorbid conditions had almost 18 times higher risk of having any ADL disability (odds ratio [OR]=18.8; 95% confidence interval [CI], 3.3-105.2) and 10 times higher risk of having any IADL disability (OR=10.6; 95% CI, 1.1-101.5), compared with subjects without either of the 2 conditions. The risk of disability was further increased if the subject had a comorbid condition (hypertension, heart attack, cancer, hip fracture, arthritis). The risk of fair or poor self rated health was 3.5 (95% CI, 1.4-8.6) and the hazard ratio for 5-year mortality was 2.4 (95% CI, 1.7-3.4) in people with both diseases. CONCLUSIONS: Diabetes and stroke in combination is strongly associated with a higher risk of disabilities, poor self-rated health, and 5-year mortality in elderly Mexican Americans. The effect on outcomes appears to follow an additive model. Information on disability risk and morbidity and mortality should be useful to rehabilitation professionals in discharge planning and allocation of therapy resources. PMID- 12736890 TI - Validation of the Berg Balance Scale as a predictor of length of stay and discharge destination in stroke rehabilitation. AB - OBJECTIVE: To validate the utility of the Berg Balance Scale (BBS) in predicting length of stay (LOS) and discharge destination for patients admitted to a stroke rehabilitation unit. DESIGN: Prospective study. SETTING: Provincial tertiary inpatient stroke unit for a primarily geriatric population. PARTICIPANTS: A total of 313 of the 325 patients admitted consecutively between April 1998 and August 2000. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: LOS and discharge destination. RESULTS: Admission BBS scores correlated negatively with LOS (r= .53, controlling for age). Logistic regression confirmed that the following were independent predictors of being discharged home rather than to an institution (adjusted odds ratio, 95% confidence interval): admission BBS (1.09, 1.06-1.12) and the presence of family supports (15.0, 7.2-31.3). These results generally concur with previously published results, obtained at a different stroke rehabilitation setting. CONCLUSIONS: This study validates the use of the BBS scores in assisting to estimate approximate LOS and eventual discharge destination. Age did not correlate significantly with the outcomes measured in this study, which was conducted in a geriatric population. PMID- 12736891 TI - Static magnetic field therapy for symptomatic diabetic neuropathy: a randomized, double-blind, placebo-controlled trial. AB - OBJECTIVE: To determine if constant wearing of multipolar, static magnetic (450G) shoe insoles can reduce neuropathic pain and quality of life (QOL) scores in symptomatic diabetic peripheral neuropathy (DPN). DESIGN: Randomized, placebo control, parallel study. SETTING: Forty-eight centers in 27 states. PARTICIPANTS: Three hundred seventy-five subjects with DPN stage II or III were randomly assigned to wear constantly magnetized insoles for 4 months; the placebo group wore similar, unmagnetized device. INTERVENTION: Nerve conduction and/or quantified sensory testing were performed serially. MAIN OUTCOME MEASURES: Daily visual analog scale scores for numbness or tingling and burning and QOL issues were tabulated over 4 months. Secondary measures included nerve conduction changes, role of placebo, and safety issues. Analysis of variance (ANOVA), analysis of covariance (ANCOVA), and chi-square analysis were performed. RESULTS: There were statistically significant reductions during the third and fourth months in burning (mean change for magnet treatment, -12%; for sham, -3%; P<.05, ANCOVA), numbness and tingling (magnet, -10%; sham, +1%; P<.05, ANCOVA), and exercise-induced foot pain (magnet, -12%; sham, -4%; P<.05, ANCOVA). For a subset of patients with baseline severe pain, statistically significant reductions occurred from baseline through the fourth month in numbness and tingling (magnet, -32%; sham, -14%; P<.01, ANOVA) and foot pain (magnet, -41%; sham, -21%; P<.01, ANOVA). CONCLUSIONS: Static magnetic fields can penetrate up to 20mm and appear to target the ectopic firing nociceptors in the epidermis and dermis. Analgesic benefits were achieved over time. PMID- 12736893 TI - Gait rehabilitation in a patient affected with Charcot-Marie-Tooth disease associated with pyramidal and cerebellar features and blindness. AB - Charcot-Marie-Tooth (CMT) disease, an inherited neuropathy characterized by length-dependent degeneration of the motor and sensory nerve fibers with consequent distal muscle atrophy and sensory reduction, can be associated with symptoms and signs of involvement of the central nervous system and/or cranial nerves. We present a patient with relatively severe CMT, cerebellar ataxia, pyramidal involvement, and blindness due to Leber's hereditary optic neuropathy. The patient presented with poor standing and gait, with consequent severe disability. Factors responsible for the patient's functional impairment (plantarflexor failure, footdrop, foot rotation, knee flexor contracture, poor proprioception, cerebellar dysfunction, spastic paraparesis, blindness) were identified and addressed by a rehabilitation management, which included, as a main intervention, ankle stabilization by drop-foot boots instead of ankle-foot orthoses. Improved balance and independent ambulation resulted from rehabilitation. PMID- 12736892 TI - Epidemiology of limb loss and congenital limb deficiency: a review of the literature. AB - OBJECTIVE: To examine the state of research on population-based studies of the incidence of limb amputation and birth prevalence of limb deficiency. DATA SOURCES: A total of 18 publication databases were searched, including MEDLINE, CINAHL, and the Cochrane Library. STUDY SELECTION: The search was performed by using a hierarchical process. Articles were reviewed for inclusion by 3 reviewers. Inclusion criteria included defined catchment area, calculation of population-based incidence rates, defined etiology of limb loss, and English language. Review articles, animal studies, case reports, cohort studies, letters, and editorials were excluded. DATA EXTRACTION: Figures on the estimated incidence of amputation and birth prevalence of congenital limb deficiency were gleaned from selected reports and assembled into a table format by etiology. DATA SYNTHESIS: The studies varied in scope, quality, and methodology, making comparisons between studies difficult. Incidence rates of acquired amputation varied greatly between and within nations. Rates of all-cause acquired amputation ranged from 1.2 first major amputations per 10,000 women in Japan to 4.4 per 10,000 men in the Navajo Nation in the United States between 1992 and 1997. Consistent among all nations, the risk of amputation was greatest among persons with diabetes mellitus. CONCLUSIONS: Surveillance of congenital limb deficiency exists in much of the developed world. Existing studies of acquired amputation suffer from a host of methodologic problems. Future efforts should be directed toward the application of standardized measures and methods to enable trends to be evaluated over time and comparisons to be made within and between countries. PMID- 12736894 TI - An unusual cause of ischemic claudication: a case report. AB - A 56-year-old woman with a chief complaint of left lower-extremity numbness was referred by her gynecologist to the physical medicine clinic for workup of presumed lumbosacral radiculopathy. She had no history of low back pain, and her symptoms were elicited only with exercise. Results of her neurologic examination and lumbosacral radiographs were normal. Her medical history was significant for advanced cervical cancer, successfully treated with local surgery followed by high-dose pelvic radiation and chemotherapy 2 years before the current onset of symptoms. Subsequent workup with Doppler and arteriogram studies discovered a 3 cm area of diffuse stenosis of the left external iliac artery for which she was successfully treated with balloon angioplasty. This case presents an unusual cause of left leg claudication secondary to left iliac artery stenosis 2 years after pelvic radiation for cervical cancer and shows the necessity for a detailed evaluation of patients' medical histories. PMID- 12736895 TI - Aerophagia as a cause of ineffective phrenic nerve pacing in high tetraplegia: a case report. AB - We report an unusual case of aerophagia after traumatic spinal cord injury (SCI), which shows the profound effects of abdominal distension on respiratory ability in such individuals. In this case, abdominal distension resulting from aerophagia reduced the effectiveness of phrenic nerve pacing on diaphragm function necessitating greater use of positive-pressure ventilatory (PPV) support. Reduction of postprandial gastric air and abdominal distension with insertion of a percutaneous endoscopic gastrostomy tube ameliorated the condition and allowed for more effective phrenic nerve pacing and greater PPV-free breathing. We are unaware of a similar case involving an individual with an SCI. PMID- 12736896 TI - Bursitis, adventitious bursa, localized soft-tissue inflammation, and bone marrow edema in tibial stumps: the contribution of magnetic resonance imaging to the diagnosis and management of mechanical stress complications. AB - OBJECTIVE: To assess the contribution of magnetic resonance imaging (MRI) in the diagnosis of tibial stump bursitis, in the establishment of differential diagnosis, and in the therapeutic management prosthetic-stump interface, mainly by adaptation of the prosthetic device. DESIGN: Two-year, prospective, consecutive series. SETTING: University-affiliated prosthetic and rehabilitation center and university department of radiology. PARTICIPANTS: A group of 17 persons with stump problems identified from a total of 139 consecutive below-knee amputees with prosthesis problems. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Clinical symptoms and MRI. RESULTS: Clinical symptoms (variable stump volume, fluctuating mass at palpation with or without mechanical pain) were suggestive of bursitis in 10 patients. MRI confirmed bursitis in 9 and identified 1 in whom clinical signs suggested neuroma, giving an incidence of 10 of 139 amputees (7.2%). MRI identified 13 sites of bursitis (adventitious bursa, 11; synovial bursitis, 2) and 5 localized areas of soft tissue inflammation. MRI showed diffuse muscular edema at 1 site of clinically suspected bursitis, and bursitis at another site of suspected neuroma. Calcified bursitis was observed in 1 case. Bone abnormalities associated with bursitis (n=7) included osteophytes or fracture (n=4) or bone marrow edema (n=3). Two asymptomatic neuromas were also identified. MRI-guided modifications of the prosthetic interface led to favorable outcome in all cases. CONCLUSION: Bursitis, adventitious bursae, and areas of localized soft-tissue inflammation are different aspects of the same disorder resulting from a mechanical conflict between the stump and the prosthesis socket. Besides contributing to diagnosis, MRI provides a precise assessment necessary for correcting the prosthesis-stump interface in a way that reduces mechanical stress and subsequently cures bursitis. PMID- 12736897 TI - Arachidonic acid cytotoxicity: can arachidonic acid be a physiological mediator of cell death? AB - Arachidonic acid is a polyunsaturated fatty acid that mediates inflammation and the functioning of several organs and systems either directly or upon its conversion into eicosanoids. However, arachidonic acid is found to be cytotoxic in vitro at concentrations that overlap physiological ones. It is tempting therefore to speculate that arachidonic acid may be a physiological inducer of apoptosis and that such cytotoxic action may be another of its roles in vivo. Nevertheless its pro-inflammatory and oxidative stress-inducing features are characteristic of necrosis and pathological conditions. We hereby review the cytotoxic action of arachidonic acid, indicate the possible pathways that lead to cell death and contemplate the cytotoxic role of arachidonic acid in vivo. PMID- 12736898 TI - Effect of dexamethasone on neutrophil metabolism. AB - The effect of dexamethasone on glucose and glutamine metabolism was investigated. The consumption and oxidation of glucose and glutamine, and the production of glutamate and lactate were determined in neutrophils cultured for 3 h in the presence of dexamethasone. The activities and expression of glucose-6-phosphate dehydrogenase (G6PDH) and phosphate-dependent glutaminase were also determined under the same conditions. Addition of dexamethasone to the culture medium caused a significant increase of glucose consumption at 0.5 microm (123.9%) and 1.0 microm (78.3%) concentrations. In spite of this, however, glucose oxidation remained unchanged. The glucocorticoid did not change glutamine consumption but caused a significant increase of glutamate production and did not alter glutamine oxidation. Dexamethasone-treated neutrophils had a significant decrease of G6PDH activity and expression in particular at 1.0 microm concentration. Phosphate- dependent glutaminase activity was also decreased (about 34%) by dexamethasone treatment. A similar effect was observed on glutaminase expression as indicated by RT-PCR analysis. Thus, the effect of dexamethasone on neutrophil metabolism was particularly noticeable with respect to G6PDH and glutaminase activities where a decrease in the respective mRNA levels was demonstrated. PMID- 12736899 TI - Chemotactic activity of oligopeptides containing an EWS motif on Tetrahymena pyriformis: the effect of amidation of the C-terminal residue. AB - Chemotactic properties of 3-7-mer peptides containing an EWS motive and their peptide amides synthesized and characterized by us were investigated in Tetrahymena pyriformis GL model. Analysis of the peptide acids shows that SEWS possesses exceptionally strong (660%+/-21; 430%+/-18) chemoattractant ability at 10(-12) and 10(-11) m respectively. The shorter peptide (EWS) possesses chemorepellent activity, while longer peptides display neutral (WSEWS) or moderate chemoattractant (EWSEWS and GEWSEWS) chemotactic ability. Amidation of the C-terminus can significantly modify the character of peptides: it points to the conclusion that a free alpha-COOH group at this position is required for the high efficiency of SEWS, while in the shorter (EWS) and longer peptides (WSEWS and EWSEWS) amidation can result in chemoattractant ligands. Evaluation of the structure-function relationship of these compounds establishes the significance of Glu (E) with its high surface-exposed area and negatively-charged side chain. The high discriminative ability and good chemotactic responsiveness of Tetrahymena support the theory that a chemotactic signalling mechanism working in higher levels of phylogeny is a well conserved and inducible one even in protozoa. PMID- 12736900 TI - Effects of melatonin on oxidative-antioxidative status of tissues in streptozotocin-induced diabetic rats. AB - In view of the antioxidant properties of melatonin, the effects of melatonin on the oxidative-antioxidative status of tissues affected by diabetes, e.g. liver, heart and kidneys, were investigated in streptozotocin (STZ)-induced diabetic rats in the present study. Concentrations of malondialdehyde (MDA) and reduced glutathione (GSH), and activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the tissues were compared in three groups of 10 rats each (control non-diabetic rats (group I), untreated diabetic rats (group II) and diabetic rats treated with melatonin (group III)). In the study groups, diabetes developed 3 days after intraperitoneal (i.p.) administration of a single 60 mg kg(-1) dose of STZ. Thereafter, while the rats in group II received no treatment, the rats in group III began to receive a 10 mg kg(-1) i.p. dose of melatonin per day. After 6 weeks, the rats in groups II and III had significantly lower body weights and higher blood glucose levels than the rats in group I (p < 0.001 and p < 0.001, respectively). MDA levels in the liver, kidney and heart of group II rats were higher than that of the control group (p < 0.01, p < 0.05, p < 0.01, respectively) and diabetic rats treated with melatonin (p < 0.05). The GSH, GSH Px and SOD levels increased in diabetic rats. Treatment with melatonin changed them to near control values. Our results confirm that diabetes increases oxidative stress in many organs such as liver, kidney and heart and indicate the role of melatonin in combating the oxidative stress via its free radical scavenging and antioxidant properties. PMID- 12736901 TI - Dantrolene protects erythrocytes against oxidative stress during whole-body irradiation in rats. AB - In our study, we examined the radioprotective effects of dantrolene against gamma irradiation-induced damage of blood cells after total body irradiation of rats. Rats were divided into three groups of eight rats each. The first group was the control group receiving no dantrolene or irradiation, the second group received total body irradiation (RT) with 5 Gy of gamma irradiation only, and the third group received dantrolene at a dose of 5 mg x kg(-1) plus RT. Dantrolene was given intraperitoneally 30 min before RT. All groups were sacrificed 2 h after RT, and blood samples were taken. Leukocyte, and thrombocyte counts and hemoglobin levels were measured. Furthermore, malondialdehyde (MDA) levels in plasma and erythrocytes and superoxide dismutase (SOD) and glutathione peroxidase activities (GSH-Px) in erythrocytes were determined. It was found that pretreatment with dantrolene at a dose of 5 mg x kg(-1) significantly reduced the MDA levels and increased the antioxidant SOD and GSH-Px activities, and prevented the decrease in leukocyte and thrombocyte counts. We conclude that dantrolene has clear antioxidant properties when given prior to radiation exposure and the protective effect of dantrolene against damage inflicted by radiation, depends, at least in part, on the decrease in lipid peroxidation and increase in the activity of antioxidant enzymes SOD and GSH-Px. PMID- 12736902 TI - Investigation of antioxidant vitamins (A, E and C) and selenium levels in chickens receiving estrogen or testosterone. AB - In the present study estrogen or testosterone was administered to broiler chickens (6 weeks old) for 5 weeks and levels of antioxidant vitamins (A, E and C) and selenium (Se) were determined. In animals who received estrogen, vitamins A, E, C and Se levels were 0.70 +/- 0.19, 11.0 +/- 2.45, 20.0 +/- 5.17 and 130.0 +/- 25.0 microg l(-1), respectively. Vitamins A, E, C and Se levels in the testosterone-administered group were found to be 0.54 +/- 0.16, 9.9 +/- 1.96, 18.0 +/- 5.18 and 100.0 +/- 18.0 microg l(-1), respectively. Vitamins A, E, C and Se levels were found to be significantly increased in the estrogen-administered group compared to the controls (p < 0.01, p < 0.01, p < 0.05, p < 0.05, respectively). Although all parameters were increased in testosterone-treated animals, only increases in vitamins A and E were found to be statistically significant (p < 0.01, p < 0.01, respectively). Based on the present findings, estrogen and testosterone show direct antioxidant effects by increasing the activities of some enzymes and they also cause an increase in antioxidant vitamin levels and hence indirectly also contribute to antioxidant capacity. PMID- 12736903 TI - Protective effects of antioxidants on age-related changes in the electrophoretic patterns of cardiac LDH, hepatic ALP and serum proteins in male golden hamster. AB - Basal and antioxidant-induced changes in the isoenzyme and isoform patterns of cardiac lactate dehydrogenase (EC 1.1.1.27) and hepatic alkaline phosphatase (EC 3.1.3.1), respectively, as well as the electrophoretic patterns of serum proteins in different age groups of male golden hamster were compared. This is to test whether age-induced changes could be corrected by long-term administration of antioxidants. Data indicated that aging causes no remarkable change in the total activity of either cardiac LDH or hepatic ALP, however a significant increase in the fractional activity of some cardiac LDH isoenzymes and a significant reduction in the fractional activity of some hepatic ALP isoforms were induced by aging. On the other hand, long-term administration of antioxidants appeared to manifest a clear counteracting effect on the age-related changes in old age. This effect was indicated in the fractional activity of cardiac LDH isoenzymes and of hepatic ALP isoforms. The present study has also shown a wide-range variation in serum protein patterns due to aging and/or antioxidant administration, which indirectly reflect a parallel variation in the process of gene expression and/or proteolytic activity. PMID- 12736904 TI - Sodium selenite modulates tumour marker indices in N-nitrosodiethylamine initiated and phenobarbital-promoted rat liver carcinogenesis. AB - The effect of sodium selenite (Se) was investigated against two-stage rat liver carcinogenesis initiated by a single intraperitoneal injection of N nitrosodiethylamine (DEN, 200 mg kg(-1) i.p.) followed by promotion with phenobarbital (PB, 0.05%) in a basal diet. Se (4 p.p.m.) was administered per os daily throughout the entire experiment, before the initiation, or during the promotion stage. The plasma, liver (hepatoma and surrounding tissue) and kidney tissue were investigated biochemically for lactate dehydrogenase, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and 5' nucleotidase. These enzyme activities were increased (p < 0.001) in plasma of hepatoma-bearing rats compared with normal control rats. The elevation of these enzyme activities in plasma was indicative of the persistent deteriorating effect of DEN in cancer-bearing animals. Aminotransferase levels were decreased in hepatoma and surrounding liver tissue, whereas lactate dehydrogenase, alkaline phosphatase and 5'-nucleotidase were increased in the cancer condition. These enzyme activities were reversed to near normal control values in animals treated with Se. It is apparent that the beneficial effect of Se is primarily exerted on the initiation phase and secondarily during the promotion stage of DEN-initiated rat liver carcinogenesis. The analysis of marker enzyme activities taken together with our previous findings clearly indicates the antitumour efficacy of sodium selenite on DEN-induced hepatoma animals. PMID- 12736905 TI - Experimental researches on the role of phosphoinositide-specific phospholipase C in 4-hydroxynonenal induced exocytosis. AB - The action of 4-hydroxynonenal (HNE), a chemotactic aldehyde produced by lipid peroxidation, was analysed on exocytosis in parallel with its effects on phosphoinositide-specific phospholipase C (PLC) both in undifferentiated HL-60 cells and in cells induced to differentiate toward the granulocytic cell line by 1.25% DMSO. Exocytosis was evaluated by the secretion of beta-glucuronidase from cells incubated at 37 degrees C for 10 min in the presence of various aldehyde concentrations. HNE action was more pronounced in DMSO-differentiated cells, where concentrations between 10(-8) and 10(-6) m were able both to trigger exocytosis and to strongly activate PLC; in both processes maximal stimulation was given by 10(-7) m. HNE-induced exocytosis was completely prevented by pertussis toxin and by the PLC inhibitor U73122. The comparison between HNE and formyl-methionyl-leucyl-phenylalanine (fMLP), used as a positive control, showed that the tripeptide produced an higher stimulation of exocytosis than the aldehyde; by contrast HNE induced a stronger increase of PLC activity. Wortmannin, an inhibitor of phosphatidylinositol-3-kinase (PI3K), strongly inhibited the exocytosis induced by fMLP, while it failed to induce a statistically significant inhibition of HNE action. We conclude that both compounds trigger exocytosis through a Ptx-sensitive G protein; the present data support the hypothesis that the lower ability of the aldehyde to trigger exocytosis as compared to fMLP might depend upon a low ability to activate PI3K, while PLC activation appears to play a key role in HNE-induced exocytosis. PMID- 12736906 TI - Trypsin induces tumour necrosis factor-alpha secretion from a human leukemic mast cell line. AB - Trypsin activating both proteinase-activated receptor (PAR) 2 and PAR4 plays an important role in inflammation. We have investigated the potential of trypsin to induce TNF-alpha secretion from the human leukemic mast cell line (HMC-1). HMC-1 cells co-express both PAR2 and PAR4, and their agonist trypsin signals to HMC-1 cells. Trypsin (100 nm), SLIGKV-NH(2) (100 microm, corresponding to the PAR2 tethered ligand), or GYPGQV-NH(2) (100 microm, corresponding to the PAR4 tethered ligand) induced tumour necrosis factor (TNF)-alpha secretion from HMC-1 cells. TNF-alpha secretion by trypsin was significantly blocked by pretreatment with 50 microm PD098059, MEK-1 inhibitor. Furthermore, trypsin stimulated the activation of extracellular signal-regulated kinase (ERK) in HMC-1 cells without any detectable activation of c-Jun N-terminal kinase (JNK) and p38 MAP kinase homologue. These results show that trypsin may induce TNF-alpha secretion following activation of ERK via both PAR2 and PAR4 on HMC-1 cells. PMID- 12736907 TI - Effects of indomethacin on the divisional morphogenesis and cytoskeleton dependent processes of Tetrahymena. AB - Indomethacin (0.1 mM) causes significantly altered phospholipid synthesis in Tetrahymena and is able to influence the inositol phospholipid signalling system (9). In the present study the effects of indomethacin on the course of cell division, cyclin expression, the cortical microtubular system and on cytoskeleton dependent processes (motility, phagocytosis) were investigated. As expected from its interference with the synthesis of phospholipids, indomethacin affected Tetrahymena in a number of ways: the structure of the cortical microtubular system became irregular; in many cells the stomatogenesis (development of new oral apparatus) and the development of the fission furrow was not accompanied by elongation of the macronucleus, which is a typical phenomenon of the normal course of mitosis: apparently indomethacin uncouples these phenomena. After indomethacin treatment, the expression of both cyclin A and cyclin B(1) were reduced significantly. The cell growth rate, motility and phagocytotic activity were all considerably reduced. There are probably additional mechanisms responsible for the effect of indomethacin on the systems that control divisional morphogenesis, for microtubule-dependent processes and for the connection between nuclear and cortical alterations during the cell cycle. Effects on protein phosphorylation/dephosphorylation, on cyclin expression and on microtubular functions are probably involved. These possibilities are discussed. PMID- 12736909 TI - The inhibitory effect of BSP-A1/-A2 on protein kinase C and tyrosine protein kinase. AB - Bovine seminal plasma contains a group of similar proteins, namely BSP-A1, BSP A2, BSP-A3, and BSP-30-kDa (collectively called BSP proteins), and they are secreted by the seminal vesicles. In our study, we purified the BSP-A1/-A2 through affinity chromatography and found for the first time that BSP-A1/-A2 can inhibit the activity of protein kinase C (PKC) and tyrosine protein kinase (TPK). The inhibition was dose dependent. When the PKC and TPK activities are expressed as the logarithm of percentage activity taking the activity in the absence of the BSP-A1/-A2 as 100%, there is a linear relationship between the their activities and the dose of BSP-A1/-A2. PMID- 12736908 TI - Effects of vitamin E on microsomal Ca(2+) -ATPase activity and calcium levels in streptozotocin-induced diabetic rat kidney. AB - Vitamin E treatment has been found to be beneficial in preventing or reducing diabetic nephropathy. Increased tissue calcium and abnormal microsomal Ca(2+) ATPase activity have been suggested as contributing factors in the development of diabetic nephropathy. This study was undertaken to test the hypothesis that vitamin E reduces lipid peroxidation and can prevent the abnormalities in microsomal Ca(2+)-ATPase activity and calcium levels in kidney of streptozotocin (STZ)-induced diabetic rats. Male rats were rendered diabetic by a single STZ injection (55 mg x kg(-1) i.p.). After diabetes was verified, diabetic and age matched control rats were untreated or treated with vitamin E (400-500 IU kg(-1) x day(-1), orally) for 10 weeks. Ca(2+)-ATPase activity and lipid peroxidation (MDA) were determined spectrophotometrically. Blood glucose levels increased approximately five-fold (> 500 mg x dl(-1)) in untreated-diabetic rats but decreased to 340+/-27 mg x dl(-1) in the vitamin E treated-diabetic group. Kidney MDA levels did not significantly change in the diabetic state. However, vitamin E treatment markedly inhibited MDA levels in both control and diabetic animals. Ca(2+)-ATPase activity was 0.483+/-0.008 U l(-1) in the control group and significantly increased to 0.754+/-0.010 U l(-1) in the STZ-diabetic group (p < 0.001). Vitamin E treatment completely prevented the diabetes-induced increase in Ca(2+)-ATPase activity (0.307+/-0.025 U l(-1), p < 0.001) and also reduced the enzyme activity in normal control rats. STZ-diabetes resulted in approximately two-fold increase in total calcium content of kidney. Vitamin E treatment led to a significant reduction in kidney calcium levels of both control and diabetic animals (p < 0.001). Thus, vitamin E treatment can lower blood glucose and lipid peroxidation, which in turn prevents the abnormalities in kidney calcium metabolism of diabetic rats. This study describes a potential biochemical mechanism by which vitamin E supplementation may delay or inhibit the development of cellular damage and nephropathy in diabetes. PMID- 12736910 TI - Association between Apo B signal peptide gene polymorphism and NIDDM. PMID- 12736911 TI - Prolonged effect of a single serotonin treatment in adult age on the serotonin and histamine content of white blood cells and mast cells of rats. AB - Hormonal imprinting was provoked by serotonin treatment in adult age. Three weeks after treatment with 100 microg serotonin, the serotonin and histamine content of peritoneal cells (mast cells, lymphocytes and the monocyte-macrophage-granulocyte group), white blood cells (lymphocytes, granulocytes and monocytes) and thymic lymphocytes was studied by flow cytometry. The content of both amines was significantly higher in the mast cells of males and lower in females. Blood lymphocytes contained a higher serotonin and histamine level in males, and a lower serotonin level in females. The peritoneal monocyte-macrophage-granulocyte group contained less serotonin in both males and females. Thymocytes contained higher levels of both amines in females and higher histamine level in males. The experiments demonstrate that a single treatment at adult age can provoke imprinting, which alters-in the present case-the serotonin and histamine content of immune cells durably. PMID- 12736912 TI - Endodontic retreatment: evaluating success and dealing with failures. AB - The outcome of endodontic treatment is not always easy to predict or assess. The ability to recognize success and evaluate the causes of failure is essential to effective root canal treatment. Utilizing recent advances in technique renders retreatment of failing cases very feasible and highly successful. These advances should also reduce the incidence of failure. PMID- 12736913 TI - The effect of frequency of toothbrushing on oral health of 14-16 year olds. AB - The aim of the present study was to evaluate the impact of toothbrushing frequency on dental plaque, caries and periodontal condition in 14-16 year old students. A total of 2083 students selected from 20 schools enrolling 8th and 9th grades were investigated by a questionnaire and a clinical examination. All participants were examined for oral hygiene, dental caries and periodontal condition using Silness & Loe plaque index (Pl.l), decayed, missing and filled teeth/surfaces (DMFT/S) indices and Community Periodontal Index of Treatment Needs (CPITN), respectively. It was revealed that about 49 per cent of males and 89 per cent of the females reported to brush their teeth on a regular basis. Such differences in toothbrushing frequency between the sexes were statistically significant (P = 0.0001). The mean (Pl.l) scores were lower in those who brushed than those who did not brush their teeth, with a significantly lower scores in females than in males (P = 0.0001). There were slight but nonsignificant differences in caries experience (MFT/S) amongst males and females as related to toothbrushing frequency (P = 0.121, 0.208 respectively). While bleeding on probing (43.0 per cent) was most prevalent in students who did not brush. Calculus scores were similar in all groups. The occurrence of shallow and deep pockets in students who brushed or didn't brush their teeth were minimal (6.6-8.4 per cent). The oral health status among those who did not brush or brushed their teeth on regular or irregular basis was found to be poor and slightly varied. Therefore, more emphasis should be placed on proper oral hygiene. Also, implementation of school based oral health promotion and prevention programs is urgently needed. PMID- 12736914 TI - Public health notification: diathermy interactions with implanted leads and implanted systems with leads. PMID- 12736915 TI - A patient's own power of sucking in order to drain dental abscesses with good results. PMID- 12736916 TI - Estimates of diagnostic accuracy efficacy: how well can this test perform the classification task? PMID- 12736917 TI - Recurrent gallstone ileus: case report. PMID- 12736918 TI - Computed tomography of acute small bowel obstruction: pictorial essay. AB - In many patients, the diagnosis of SBO can often be made by a combination of clinical history, physical examination and plain radiographs. However, in many cases, the diagnosis of SBO may be difficult. Abdominal radiographs are limited in their ability to diagnose SBO and, more particularly, to identify a specific underlying cause. Although barium studies, ultrasonography and magnetic resonance imaging may all play a role in the evaluation of SBO, CT should be the examination of choice for most patients when the diagnosis and underlying cause of SBO is unclear. It may help differentiate pseudo-obstruction from true obstruction, and a specific underlying cause can often be determined. Signs of closed-loop obstruction, strangulation, perforation or infarction may be detected and allow for more timely and appropriate surgical management. We advocate the use of CT in any patient with SBO where the cause or diagnosis is unclear. We have attempted to demonstrate a range of clinical cases in our practice where CT played an invaluable role in the evaluation of our patients. PMID- 12736919 TI - Utility of a computed tomography oral imaging software program for preoperative evaluation of head and neck cancers. PMID- 12736921 TI - Primary epidural Ewing's sarcoma: case report and review of literature. PMID- 12736920 TI - On-call services provided by radiology residents in a university hospital environment. AB - OBJECTIVE: To better understand the consultative role of the radiology resident after hours. METHODS: Data were collected prospectively from Mar. 15, 1999, to Jan. 5, 2001, during on-call coverage hours at our university hospital. Urgent radiologic examinations for which the on-call resident rendered a preliminary interpretation were included in our analysis, with the following entered into a database: patient demographics, consultative time and weekday, imaging modality, consulting clinical service and indication for each study. RESULTS: A total of 1784 studies were performed on 1451 patients; most were requested by the emergency department (844 cases [47.3%]). The mean number of radiographic studies performed was 20.1 (standard error of the mean [SEM] 1.1) per weekday (n = 44) and 49.4 (SEM 1.8) per weekend day or holiday (n = 18). There were 1227 (68.8%) computed tomographic (CT), 338 (18.9%) ultrasonographic, 98 (5.5%) plain radiograph, 63 (3.5%) nuclear medicine, 21 (1.2%) interventional, 20 (1.1%) fluoroscopic and 17 (1.0%) magnetic resonance imaging examinations. The 3 most common studies were CT of the head in 692 cases, CT of the abdomen in 230 and venous Doppler ultrasonography in 158. CONCLUSIONS: Radiology residents are performing a diverse and increasing number of emergent diagnostic examinations after hours. It is therefore important that radiology departments are aware of these consultative needs to best ensure that appropriate resident skills are developed to meet these demands. PMID- 12736922 TI - Sonographic appearance of cerebral vascular air embolism in neonates: report of two cases. PMID- 12736923 TI - Ultrasound-guided thrombin injection of femoral artery pseudoaneurysms. AB - OBJECTIVE: To evaluate whether the positive initial results for ultrasound-guided percutaneous thrombin injection of pseudoaneurysms, reported predominantly in small retrospective series, would be supported in a larger prospective trial. METHODS: In April 1999, our institution adopted ultrasound-guided thrombin injection as the initial treatment for post-catheterization arterial pseudoaneurysm. Colour Doppler imaging delineates the pseudoaneurysm, its neck and the adjacent artery. A 22-gauge spinal needle is attached to a 1-mL syringe preloaded with thrombin at a concentration of 1000 U/mL. Under ultrasound guidance, the needle tip is positioned within the pseudoaneurysm, and real-time colour Doppler imaging is used to monitor the pseudoaneurysm as thrombin is slowly injected. Thrombus formation commences almost immediately, and in most cases, occlusion is complete within 5 seconds. RESULTS: We successfully treated 61 pseudoaneurysms in 61 consecutive patients. The amount of thrombin injected ranged from 20 U to 3000 U (mean 435 U); 55 pseudoaneurysms were successfully treated after a single injection, and 6 patients required a repeat injection for complete occlusion. One patient had 2 pseudoaneurysms treated on consecutive days, and 1 developed a symptomatic vasovagal reaction, which was treated conservatively. No other significant procedural complications were encountered. Fifty-nine patients had a follow-up groin Doppler sonogram between 1 and 5 days after treatment. CONCLUSION: Ultrasound-guided percutaneous thrombin injection is an effective, simple, fast and safe treatment for post-catheterization arterial pseudoaneurysm. It has replaced ultrasound-guided compression repair at our institution and is now our treatment of choice. PMID- 12736924 TI - Noninvasive imaging of carotid artery disease: critically appraised topic. PMID- 12736925 TI - Answer to case of the month #88. Spontaneous intracranial hypotension. PMID- 12736926 TI - Another opportunity for excellence. PMID- 12736928 TI - Using lidocaine for peripheral i.v. insertions: patients' preferences and pain experiences. AB - Patients' pain experiences with peripheral i.v. insertions, with and without the use of lidocaine for anesthesia, were described and compared. Patients reported their preferences related to the use of intradermal lidocaine with future i.v. insertions. PMID- 12736927 TI - Applying airline safety practices to medication administration. AB - Medication administration errors (MAE) continue as major problems for health care institutions, nurses, and patients. However, MAEs are often the result of system failures leading to patient injury, increased hospital costs, and blaming. Costs include those related to increased hospital length of stay and legal expenses. Contributing factors include distractions, lack of focus, poor communication, and failure to follow standard protocols during medication administration. PMID- 12736929 TI - Anger: the mismanaged emotion. AB - Mismanaged anger is a significant problem in health care settings. Research-based information is presented on the angry emotionality that nurses frequently encounter. Gender differences in anger are examined. Strategies are presented for dealing with angry patients, physicians, and colleagues. PMID- 12736930 TI - Laryngeal edema: perioperative nursing considerations. AB - Laryngeal edema is a relatively rare yet serious postoperative complication for those patients undergoing general anesthesia. Maintenance of airway and breathing are always the first priority of perioperative patient care. Medical-surgical nurses working in postoperative settings must be familiar with the signs, symptoms, and necessary prompt treatment for this life-threatening condition. PMID- 12736931 TI - Adult-health/medical-surgical nurses' perceptions of students' contributions to clinical agencies. AB - The nursing shortage and accompanying increase in nurses' workloads could suggest that the presence of nursing students on clinical units would further increase nurses' responsibilities. Counterbalancing this concern is the need to adequately prepare the next generation of dedicated and skilled nurses. The question arises as to whether nursing students are an asset or liability to clinical agencies. PMID- 12736933 TI - Predicting pressure ulcer sore risk. PMID- 12736932 TI - Maintaining management during disaster. PMID- 12736936 TI - Formation and technological innovation. PMID- 12736937 TI - Computer-assisted functional diagnostics: in or out? PMID- 12736938 TI - Computer-assisted instrumental functional diagnostics--state of development, possibilities, and limits. AB - The state of development, current trends, and future prospects of computer assisted instrumental functional diagnostics are discussed, including applications in articulator adjustment, determining and displaying movement function, screening where dysfunction is suspected, and more. PMID- 12736939 TI - Computer assistance in clinical functional analysis. AB - The use of computers in the dental practice has been primarily restricted to the acquisition of billing data. Additional possibilities for use of PCs exist in diagnostic data acquisition and evaluation; clinical functional analysis seems a particularly suitable application. Such software is now available: CMDfact. Dentally, it is based on a previously developed and published examination and documentation system, the graphic user interface of which is used in the newly developed software. After the examination data have been acquired by mouse click or numerical entry, these are available for evaluation. A special function, the "Diagnosis pilot" is integrated to support the user. This helps in the assignment of the appropriate "Initial diagnoses", since it brings together the individually existing principal symptoms and suitable diagnoses for the initial diagnosis in question and also states which diagnoses "would be appropriate" for this, but are not available. With 3D animation, the software also helps the dentist to explain aspects of CMD to patients. The software also assists the dentist with a detailed multimedia help system, which provides context-sensitive help for every examination step. These help functions explain the sense of the relevant examinations, their performance and evaluation in the form of short texts and explanatory photographs and videos. PMID- 12736940 TI - Cerec Scan bridgework: a two-year report. AB - This case shows a 3-unit bridge fabricated with the Cerec Scan system and a single Vita Mark II ceramic blank in service for over two years. The expanded capabilities of the Cerec 3 allow the operator to create restorations that have typically been relegated to the laboratory realm even for Cerec users. PMID- 12736942 TI - Esthetic and phonetic rehabilitation. AB - A case of crown replacement for esthetic and phonetic reasons using allceramic CAD/CAM restorations is presented. PMID- 12736941 TI - Ultimate accuracy with correlation. AB - A step-by-step guide to creating excellent occlusal surfaces using Correlation mode, C-Stat and "Anatomical Outlining" is presented. Potential pitfalls are also discussed. PMID- 12736945 TI - Asthma pathway treats and teaches patients. PMID- 12736943 TI - ED case managers keep patients from falling through the cracks. PMID- 12736944 TI - A better way to consider information technology. PMID- 12736947 TI - LOS reduced as system hones case management. PMID- 12736946 TI - Check mental status or risk missing problems. PMID- 12736948 TI - Committee seeks ways to enhance hospital revenue. PMID- 12736949 TI - Working in a personal care home. PMID- 12736950 TI - Be vigilant, nurses warned. PMID- 12736951 TI - Getting wired for interactive health communication. PMID- 12736952 TI - Childhood immunization. How knowledgeable are we? PMID- 12736955 TI - Dental education in Europe--conflicting principles? PMID- 12736953 TI - The undergraduate nurse employment project. PMID- 12736954 TI - Interview with Helene Sabourin. PMID- 12736956 TI - Current issues in dental practice management. Part 1. The importance of shared values. AB - There can be few who would argue with the notion that the nature of dental practice in the United Kingdom has changed dramatically over the last couple of decades. A variety of factors, including new clinical techniques, growing consumerism, a much greater awareness of health-related and well-being issues in the public at large, as well as a marked deregulation within the dental profession, the development of vocational training and recently mandatory lifelong learning, the growing number of females working in the profession, and an increasing reluctance of young dentists to finance dental practices have all combined to create an environment which has enabled and encouraged a move away from traditional forms of dental care delivery. Instead, there has been considerable growth in independently-funded practice and a commensurate growth in the number of practices operating under a corporate body umbrella of one form or another. Currently there are 27 corporate bodies registered with the General Dental Council (GDC) with the likelihood of more in the future given the proposed GDC review. This will no doubt take into consideration European law, under which the restriction within the Dentist's Act on the number of corporate bodies is likely to be untenable. Although they still have only a small share of the dental market--with 4% of all dentists in the UK in 1999--they have expanded rapidly from a small base. The data available at the time the paper was written indicate that the global total of fees earned from dentistry in the UK in the financial year 2001/2002 was almost 3 billion Pounds, of which 1.9 billion Pounds (64%) came from NHS fees and 1.1 billion Pounds (36%) from private fees. Of this 1.9 billion Pounds received in NHS fees in 2001/2002, 0.55 billion Pounds were paid by patients who were not exempt from charges, bringing the total amount actually paid out of patients' pockets for dental treatment to 1.65 billion Pounds. Compare these figures with 1996/1997 when NHS fees stood at 1.6 billion Pounds (71%) and private fees at 0.6 billion Pounds (29%) and it can be seen that while the size of the whole market has grown NHS fees have risen by around 18% while private sector fees have gone up by 64% during the same period. It cannot be said therefore that NHS dentistry has disappeared although it is clear that access to NHS dentistry has been and may still be difficult in some parts of the country. The Department of Health in England has responded by developing 'access centres' and in the recently published Options for Change sets out sweeping proposals to revitalise NHS dentistry and delegate decisions over funding to a local level. It is far from clear what the consequences of these changes will be. However, one possible scenario is that in many parts of the UK, although relief of pain, prevention and restoration of function will be funded by the NHS, other than in 'special circumstances' such as after trauma and inherited conditions 'cosmetic embellishment' will not be funded. In which case, ever increasing public interest in personal appearance may well lead to further growth in private dentistry. Against a background of change and uncertainty, this series of articles explores some of the issues facing dentists coming to terms with working in this brave new world. This first part examines an aspect of dental practice that is seldom discussed openly and yet is of such fundamental importance that it is the cornerstone of all successful practices and the main fault line in unsuccessful ones, namely the nature and strength of practice values. PMID- 12736958 TI - News from the dental houses. PMID- 12736957 TI - A combined approach of enamel matrix derivative gel and autogenous bone grafts in treatment of intrabony periodontal defects. A case report. AB - Enamel matrix derivative (EMD) has recently been introduced as a new modality in regenerative periodontal therapy. This case report demonstrates a combined approach in topical application of EMD gel (Emdogain) and autogenous bone grafts for treatment of intrabony defects and furcation involvement defects in a patient with chronic periodontitis. The seven-month post-surgery clinical and radiographic results were presented. The combined application of EMD gel with autogenous bone grafts in intrabony osseous defects resulted in clinically significant gain of attachment on diseased root surfaces and bone fill on radiographs. Further controlled clinical studies are required to confirm the long term effectiveness of the combination of EMD gel and autogenous bone grafts in treatment of various osseous defects in subjects with chronic periodontitis. PMID- 12736959 TI - Restorative management of the elderly patient. AB - It is predicted that geriatric patients will dramatically increase in number within our working lives. Although they are considered to demand treatment less frequently than the younger age groups, there are specific issues that need to be considered when providing dental treatment for these patients. The term gerodontics has been coined to describe this subject. The need to consider family relationships and the role of carers may play a greater part in their dental management than that of the general population. As well as oral disease, the incidence of general disease is higher in this population, as is the amount of medication prescribed to treat it; these factors often complicate dental management. Special problems that will have a high dental impact on the geriatric patient may be lack of adaptive potential, poor plaque control, root caries, periodontal breakdown and levels of toothwear. PMID- 12736960 TI - Dentists' perceptions of trends in restorative dentistry in the UK. AB - PURPOSE OF STUDY: To obtain responses to a questionnaire on likely trends in the clinical practice of restorative dentistry from a national sample of general dental practitioners (GDPs) who were principals of group practices in the UK. BASIC PROCEDURES: A panel of nine experts had been convened previously to define areas of trends in restorative dentistry in the UK and to formulate these into statements. Twenty trends were identified and framed into a 22-item questionnaire based on the trend statements. The questionnaire was sent to a national sample of 2800 principal dentists in group practices. Names of principal dentists were taken from the Yellow Pages directories, a group practice being defined as at least one principal dentist and one other practitioner. Levels of agreement for questionnaire scores of > or = 70% for all segments of a statement were taken as indicating a possible trend. MAIN FINDINGS: Of the 2800 questionnaires sent out, 1448 were returned, a return rate of 52%. Of these, 1217 questionnaires were usable, an overall response rate of 44%. Good agreement was obtained for 14 statements (64%). PRINCIPAL CONCLUSION: The 14 trend statements that scored a level of good agreement were taken as indicating possible trends in dentistry in the UK as perceived by those GDPs who responded to the questionnaire. However, in view of the fact that the 1217 usable questionnaires received reflected the opinions of 10% of the principal dentists in group practice in the UK, this will not be representative of all principal dentists in practice. Nevertheless for the purposes of gaining some insight into trends in restorative dentistry, it could be considered to be of value. PMID- 12736961 TI - Reasons for the placement and replacement of crowns in general dental practice. AB - AIMS: The purpose of the study was to apply established methods to survey reasons for the placement and replacement of crowns in general dental practice in the United Kingdom. MATERIALS AND METHODS: One hundred and twenty-eight general dental practitioners were recruited. Participants recorded the principal reason for the provision of each initial and replacement crown they provided over a 12 week period. RESULTS: Overall, data were collected from 92 practitioners in respect of 1714 patients and 2164 crowns, of which 1452 (67%) were initial placements and 712 (33%) replacements. The teeth most frequently crowned were maxillary incisors (33%), with 72% of the crowns surveyed being of the porcelain bonded to metal variety. Overall 64% of the initial placement crowns were provided because of restoration failure (26%) or tooth fracture (38%). The most common reason for crown replacement was crown failure (27%). CONCLUSION: It is concluded that surveys of the type reported may provide new insights into the reasons for and pattern of provision of initial placement and replacement crowns in clinical practice. In this study the most common reason for the provision of initial placement crowns was tooth fracture. The most common reason for the replacement of crowns, notably porcelain jacket crowns, was crown fracture. PMID- 12736962 TI - Re-inforcement of Hib immunisation required. PMID- 12736963 TI - Moving on from bureaucracy to behaviour change in food businesses. PMID- 12736964 TI - The PHLS is dead: long live the HPA. PMID- 12736965 TI - Study of cleaning standards and practices in food premises in the United Kingdom. AB - A study was undertaken to determine the microbiological status of surfaces used in the preparation of ready-to-eat foods, and to assess cleaning standards and practices in food premises in the UK. A total of 6,533 environmental samples were examined from 1,502 catering (such as restaurants, cafes, and sandwich bars) or retail premises (such as butchers, delicatessens, and bakers): 2,033 samples from chopping/cutting boards, 2,009 from worktop surfaces, 1,359 from food containers, and 1,132 from cleaning cloths. Cleaning cloths were more heavily contaminated with bacteria (Aerobic Colony Count (ACC), Enterobacteriaceae, E. coli, and Staph. aureus) compared to surfaces sampled. Campylobacter spp. were detected in two (0.2%) and Salmonella spp. from one (0.1%) of the cleaning cloths. Surfaces that were visually dirty, wet, last cleaned over 24 hours ago, and boards that were scored or damaged were found to have higher levels of bacteria. A hazard analysis system was in place in most (70%) food premises visited, and in 52% it was documented. Most managers (89%) had received some form of food hygiene training. Documented cleaning schedules and cleaning records were only present in approximately half (55% and 44%, respectively) of the premises. Most did not have separate implements for cleaning raw and ready-to-eat food areas (67%), or stored cleaning equipment for high risk (ready-to-eat food) areas away from those used in low risk (raw, non ready-to-eat food) areas (70%). Deficiencies in the correct use of cleaning products, such as the minimum contact time for disinfectants, were identified. Surface samples (chopping/cutting boards, worktops, and food containers) and cleaning cloths with ACC levels in excess of 10(3) cfu/cm2, swab or ml were associated with premises that did not have management food hygiene training, hazard analysis, cleaning schedules or cleaning records in place, and with little or no confidence in the food business management of food hygiene as indicated by Local Authority Inspectors' Confidence in Management scores. PMID- 12736966 TI - Presumed Pseudomonas folliculitis outbreak in children following an outdoor games event. AB - In early summer 2001, an outbreak of atypical rash occurred among children from 24 junior schools who attended an outdoor games event. The event comprised a series of five water games including a water slide, and within 24 hours of attending the event 151/593 (26%) children developed a papulopustular rash. The rash had a characteristic distribution, predominantly over the lower trunk and buttocks, with some involvement of the arms and legs. Pseudomonas aeruginosa was isolated from a water butt used to draw water for the games, and from the tank of a fire engine that supplied the water. Similar outbreaks, due to Pseudomonas folliculitis, have been previously described and in immunocompromised people this can be associated with serious illness. We recommend that superchlorinated water be used for water play events and that care be taken to avoid abrasive surfaces. PMID- 12736967 TI - Results of an extended tuberculosis screening programme among sixth formers in a London school--more questions than answers. AB - Following the death of an unvaccinated 16-year-old school student with isoniazid resistant pulmonary tuberculosis, extended screening of sixth formers took place to identify further cases of tuberculosis and to establish the need for BCG vaccination. Eight hundred and four 16-19 year olds in the school underwent Heaf testing and completed a demographics questionnaire. Forty-nine (5.5%) of these children had a positive Heaf test and were offered a chest radiograph. Four children were diagnosed with pulmonary or mediastinal tuberculosis, none linked to the index case. Fifty-four students (6.7%) with no prior BCG had a Heaf grade 0-1 reaction and were recommended for vaccination. Ninety-one percent of students were from ethnic minority groups and 29% had been born outside the UK. British Thoracic Society recommendations on the management of tuberculin positive cases of this is open to interpretation and we suggest that a more directed and aggressive approach to TB control should be considered in inner city schools with a high proportion of at risk ethnic minority students. PMID- 12736968 TI - Management of hepatitis A in a food handler at a London secondary school. AB - A case of hepatitis A in a food handler at a local secondary school prompted the institution of control measures including administration of human normal immunoglobulin (HNIG) to all pupils. This paper summarises the management of the incident and describes the use of HNIG to prevent secondary cases. Few adverse effects were reported and only one secondary case of hepatitis A occurred at the school. PMID- 12736969 TI - Hepatitis A virus infection control: an audit of practice in England and Wales prior to guidelines being published. AB - This survey was conducted in September 2001 to audit the practice of Consultants in Communicable Disease Control (CsCDC) and to gain a systematic picture of pre guideline approaches to the control of hepatitis A virus infection in England and Wales. An audit form was distributed to all CsCDC by email and responses were entered in a database and analysed. Response rate is estimated at 44%. CsCDC predominantly recommend vaccination for prevention. For 31% of the CsCDC, index cases were not reported within a week of the onset of illness. As a result, vaccine was often used sub-optimally. By contrast 95% of the CsCDC were informed of an index case within two weeks of onset of illness, in time for effective use of human normal immunoglobulin (HNIG). CsCDC were cautious in the use of HNIG for a number of reasons including concerns about variant Creutzfeldt-Jacob Disease (vCJD). A substantial proportion (between 5% and 28% for different age groups) of CsCDC did not recommend any active prevention among close family contacts of a case. Important ways to improve practice include facilitating use of HNIG and speeding up reporting by doctors and laboratories. PMID- 12736970 TI - Risk factors, clinical features and genotype distribution of diagnosed hepatitis C virus infections: a pilot for a sentinel laboratory-based surveillance. AB - Hepatitis C is a global public health problem. A cross-sectional survey was undertaken to determine the frequency of reported risk factors and possible transmission routes in individuals in whom HCV antibody (anti-HCV) was newly detected. Seven public health laboratories in England and Wales reported persons with positive anti-HCV tests over a three-month period (1st November 1996-31st January 1997). A questionnaire was then sent to the clinician or general practitioner (GP) who requested the test. A total of 320 laboratory reports were received from participating laboratories and 221 (69%) questionnaires were received from clinicians and GPs. Of those patients from whom a questionnaire was received (median age 36 years; males 72.9%, females 23.1%), 86% had one or more risk factors for infection reported by the clinician/GP. Injecting drug use (68%) was the main risk factor reported. Reasons for testing included being in a known risk group (65%), liver disease (19%) and blood donation (1.4%). Of the total responders, 67% were asymptomatic, and of those that had had liver function tests 50% were abnormal. The most prevalent HCV genotypes were 3a and 1a. Risk factors for HCV infection can be identified using a simple postal questionnaire to clinicians/GPs who request patient screening. PMID- 12736971 TI - Retrospective screening for hepatitis C in a tertiary paediatric referral centre. AB - The true prevalence of hepatitis C virus in children in the UK is not known and targeted screening is not standard practice despite an anticipated rise in new cases due to vertical transmission. An extension of the Department of Health's 'look-back' exercise was undertaken in order to determine the prevalence of hepatitis C virus in high-risk patient groups who were transfused with blood and/or blood products before 1991. Five hundred and ninety-five patients transfused between 1971-91 were traced and offered counselling and testing. Blood samples from 405 were analysed for the presence of HCV antibodies and/or HCV RNA by RT-PCR and eight patients were found to be positive. The HCV seroprevalence rate in this cohort was 1.97% and the HCV genome detection rate was 1.72%. In view of the long-term complications from this infection and the availability of potentially effective anti-viral agents, we feel that targeted screening is of value in this setting. PMID- 12736972 TI - Laboratory diagnosis of urinary symptoms in primary care--a qualitative study. AB - The use of the microbiology laboratory for the investigation of urinary symptoms and the test positivity rate for the urine specimens varies considerably. We used qualitative methods to explore how and why primary care staff make diagnostic and management decisions in patients presenting with urinary symptoms. Stratified random sampling was used to include general practitioner (GP) practices that were heavy and light users of the microbiology service. The data were analysed using a modified grounded theory approach utilising the constant comparative method. Frequent users of the laboratory, with low specimen positivity, did not use near patient tests (NPTs) and sent urines from most patients with urinary symptoms to confirm their clinical diagnosis. Frequent users with high positivity advise patients with negative NPTs and send all specimens with positive NPTs for antibiotic susceptibility testing. Infrequent users did not value the laboratory for diagnosis, made great use of NPTs, and tended to prescribe empirical antibiotic treatment even in patients with minimal symptoms. Guidance on the management of urinary symptoms, the use of near patient tests and laboratory culture should form part of the ongoing education of primary care staff. PMID- 12736973 TI - Survey of general practitioner satisfaction with a district communicable disease control service. AB - General practitioners (GPs) have an important role in disease surveillance and control, and are therefore a key target audience for a district communicable disease control (CDC) service. The feedback of surveillance information, and the provision of policies, guidance and specialist advice, are essential functions of a CDC service. This survey was undertaken to assess how well a CDC service was meeting the needs of GPs. Satisfaction with the service was assessed using a questionnaire survey of 166 GPs. The response rate was 42%. GPs reported a high level of overall satisfaction. The service was valued and it influenced GP practice. The service was valued more for responses to requests for information and assistance than for communicable disease control policies. The highest satisfaction was expressed for communication and for immunisation advice. Several areas of need were identified, including the need for surveillance information on antibiotic resistance and for sexually transmitted infections. The survey tool is recommended as a way of auditing standards for Good Public Health Practice, as identified by the Faculty of Public Health Medicine. PMID- 12736974 TI - Rising incidence of Haemophilus influenzae type b disease in England and Wales indicates a need for a second catch-up vaccination campaign. AB - The incidence of invasive Haemophilus influenzae type b (Hib) disease in the UK fell rapidly following the introduction of routine vaccination in 1992 and the implementation of a catch-up campaign in children under 4 years old in 1992-93. However, since 1999 the number of cases of Hib has been increasing, and in 2002 there were 134 cases in 0-4 year olds (266 in all ages). While still much less than the prevaccine burden of disease (over 800 cases a year in 0-4 year olds), this increase in incidence is worrying and has sparked a range of detailed investigations. In February 2003, the Department of Health announced a second catch-up campaign offering all children between 6 months and 4 years a further dose of Hib vaccine. The epidemiology of Hib disease in England and Wales between 1990 and 2002 is reviewed here to provide a context for this public health response. PMID- 12736975 TI - Monitoring virus strain variation following infection with VZV: is there a need and what are the implications of introducing the Oka vaccine? AB - Varicella zoster virus (VZV) is a stable virus showing relatively little variation. Nevertheless, recent data have shown there to be at least four distinct viral strains. For the most part these are geographically segregated, but in areas of the world such as the UK, where mixed populations live, there is evidence for spread of all the genotypes. Little is known about the biological differences, if any, between these strains, yet recent data have shown that even a single nucleic acid change can affect the biological behaviour of the virus. The Oka vaccine has been licensed for mass vaccination in the US and for limited use in the UK, particularly in seronegative healthcare workers. Virological surveillance is needed to support these programmes and study the effect on virus spread. Evidence for VZV superinfection of latently infected individuals with different strains, and the increasing detection of VZV in association with clinical conditions such as viral meningitis, suggest more data are needed on the transmissibility and biological properties of the virus. PMID- 12736976 TI - Importance of PCR in the diagnosis and understanding of rotavirus illness in the community. AB - Rotavirus infections pose a significant economic burden due to hospitalisation and missed workdays. Recent studies have shown that RT-PCR may detect more symptomatic cases of rotavirus infection than antigen detection methods. We compared RT-PCR, enzyme immunoassay (EIA) and an immunochromatography method (ICG) applied to 107 stool samples. The RT-PCR detected 58 positive samples whereas the EIA and ICG detected 28 and 29 positive samples respectively. The RT PCR assay detected all 17 samples found positive by both the EIA and ICG. Without sensitive diagnosis, infection control policies will be disadvantaged and the true economic burden of rotavirus underestimated. PMID- 12736977 TI - First rotavirus, now astrovirus: the evolving benefits of RT-PCR. AB - We compared RT-PCR and a newly developed amplified EIA for the detection of human astrovirus (HAstV) in 48 stool samples taken from children of less than five years of age with gastroenteritis. The RT-PCR classified more samples as positive than the EIA (18 versus 8). Without sensitive diagnosis, infection control policies will be disadvantaged and the true economic burden of HAstV underestimated. PMID- 12736979 TI - The Holy Grail of the catch-all PCR. PMID- 12736978 TI - Surveillance of meningococcal disease in South Island, New Zealand. PMID- 12736980 TI - Goodbye to all that. PMID- 12736981 TI - Role of the Health and Safety Executive in Communicable Disease and Public Health. PMID- 12736982 TI - Integration of single-agent and combination chemotherapy in the management of women with metastatic breast cancer. PMID- 12736983 TI - [Acta pediatrica de Mexico, forum to diffuse the scientific contributions in the adolescent area]. PMID- 12736984 TI - [Epidemiology of Tuberculosis in Mexico, 1981-1998. Inconsistencies between reports of the WHO and the Ministry of health]. AB - OBJECTIVE: To describe the tuberculosis morbidity and mortality trends in Mexico, by comparing the data reported by the Ministry of Health (MH) and the World Health Organization (WHO) between 1981 and 1998. MATERIAL AND METHODS: The number of cases notified in the past few years, their rates, and the trends of the disease in Mexico were analyzed. The incidence of smear-positive pulmonary tuberculosis was estimated for 1997 and 1998 with the annual tuberculosis infection risk (ATIR), to estimate the percentage of bacilliferous cases in 1997 1998. RESULTS: WHO reported more tuberculosis cases for Mexico than the MH. However, this difference has decreased throughout the years. The notification of smear-positive cases remained stable during 1993-1998. The estimated percentages of detection were 66% for 1997 and 26% for 1998 (based on ATIR of 0.5%). Tuberculosis mortality decreased gradually (6.7% per year) between 1990 and 1998, whereas the number of new cases increased, suggesting the persistence of disease transmission in the population. CONCLUSIONS: Inconsistencies between case notifications from national data and WHO were considerable, but decreased progressively during the study period. According to ATIR estimations, a considerable number of infectious tuberculosis cases are not detected. The English version of this paper is available at: http://www.insp.mx/salud/index.html. PMID- 12736987 TI - In vitro antimicrobial susceptibility in clinical isolates of Enterococcus species. AB - OBJECTIVE: To describe the antimicrobial activity of several antimicrobial agents against 97 clinical significant isolates of Enterococcus spp. MATERIAL AND METHODS: During a 2-year prospective study at Instituto Nacional de Pediatria (National Institute of Pediatrics) in Mexico City. Ninety seven strains of Enterococcus spp. (60 E faecalis and 37 E. faecium) were tested against 11 antibiotics. Susceptibility tests were performed with agar, according to the standards of the sNational Committee for Clinical Laboratory Standards (NCCLS). Isolates were screened for high-level resistance (HLR) to beta-lactams, aminoglycosides, glycopeptides and other antibiotics, as well as for vancomycin phenotypes. Differences between proportions were evaluated with chi 2 of Fisher exact fest. RESULTS: Overall resistance rates to the antibiotics tested were: 17/97 (17.5%) to penicillin, ampicillin, amoxicillin-clavulanate and imipenem. There was neither HLR nor beta-lactamase production; 74/97 (48.4%) were resistant to erythromycin; 60% to ciprofloxacin; 31/97 (32%) to gentamicin, and 55/97 (56.7%) to streptomycin. Seven strains were vancomycin-resistant enterococci (VRE), all of them identified as E. faecium; 5/7 with Van A and 2/7 with Van B phenotypes. All the isolates were susceptible to linezolid. The difference in susceptibility among species was significant. CONCLUSIONS: Mutidrug-resistant enterococci is a real problem and continuous surveillance is necessary. The microbiology laboratory is the first line of defense against the spread of multiantibiotic-resistan enterococci in the hospital environment. All the strains recovered should be tested for susceptibility to ampicillin, streptomycin, gentamicin and glycopeptides. The English version of this paper is available too at: http://www.insp.mx/salud/index.html. PMID- 12736985 TI - [Taeniasis and detection of antibodies against cysticercus among inhabitants of a rural community in Guerrero State, Mexico]. AB - OBJECTIVE: To assess the frequency of Taenia solium carriers and its relationship with human cysticercosis in a Mexican locality. MATERIAL AND METHODS: A cross sectional study was conducted in 1998, in a locality of Guerrero State, Mexico. Four hundred and three fecal samples were analyzed by enzyme-linked immunosorbent assay (ELISA) to detect Taenia sp coproantigen. Ninety two serum samples were also analyzed for antibodies against cisticerci using the immunoelectrotransfer blot assay (IET). Data were analyzed using descriptive statistics and odds ratios. RESULTS: Five of the 403 fecal samples were positive (1.2%). The adult cestode was recovered in only two people. Three (3.26%) out of the 92 serum samples that were analyzed for anticysticercus antibodies were positive. Seventeen serum samples corresponded to people living with a person positive to the coproantigen test (first group), the remaining 75 were obtained from people without a history of releasing taenia proglottids (second group). In the first group, 2 positive sera were detected (11.8%), while in the second only I was positive (1.3%) (odds ratio = 9.87, 95% confidence interval 0.64 to 295.56, p = 0.08). CONCLUSIONS: The difficulty to obtain the adult parasite in persons positive to coproantigens, may be due to difficult expulsion, a shorter permanence of the parasite in the host, insufficient dosage of treatment, or to lack of specificity of the diagnostic test. Further studies are needed to evaluate these possibilities; a better knowledge of parasite transmission dynamics will allow the implementation of prevention and control measures and a better assessment of diagnostic tests under field conditions. The English version of this paper is available at: http://www.insp.mx/salud/index.html. PMID- 12736986 TI - [Neonatal sepsis morbidity and mortality in a tertiary care hospital]. AB - OBJECTIVE: To compare the epidemiological, clinical and microbiological profiles between patients with neonatal sepsis who lived or died. MATERIAL AND METHODS: The medical records of patients with neonatal sepsis were retrospectively reviewed at Instituto Nacional de Pediatria (National Pediatric Institute) of Secretaria de Salud (Ministry of Health) in Mexico City, between 1992 and 2000. Neonatal sepsis cases were classified as surviving or not after 90 days of postnatal follow-up. The survivor and decreased groups were compared using Mann Whitney's U test for continuous variables, and the chi-squared test or the Fisher's exact test for categorical variables. Significantly associated variables were included in a Cox proportional hazards model. A p-value < 0.05 was considered statistically significant for all analyses. RESULTS: A total of 116 patients with neonatal sepsis were included (65 live and 51 dead). Multivariate analysis showed that fetal distress, respiratory distress, a delayed capillary fill up, a low platelet count, and a positive hemoculture for Klebsiella pneumoniae were significant risk factors for death. CONCLUSIONS: Epidemiological, clinical, laboratory, and microbiological variables are significant predictors of death in newborns with neonatal sepsis. The English version of this paper is available at: http://www.insp.mx/salud/index.html. PMID- 12736988 TI - [The health gap in Mexico, measured through child mortality]. AB - OBJECTIVE: To estimate the health gap in Mexico, as evidenced by the difference between the observed 1998 mortality rate and the estimated rate and the estimated rate for the same year according to social and economic indicators, with rates from other countries. MATERIAL AND METHODS: An econometric model was developed, using the 1998 child mortality rate (CMR) as the dependent variable, and macro social and economic indicators as independent variables. The model included 70 countries for which complete data were available. RESULTS: The proposed model explained over 90% of the variability in CMR among countries. The expected CMR for Mexico was 22% lower that the observed rate, which represented nearly 20,000 excess deaths. CONCLUSIONS: After adjusting for differences in productivity, distribution of wealth, and investment in human capital, the excess child mortality rate suggested efficiency problems in the Mexican health system, at least in relation to services intended to reduce child mortality. The English version of this paper is available at: http://www.insp.mx/salud/index.html. PMID- 12736990 TI - Hookworm infection and anemia in adult women in rural Chiapas, Mexico. AB - OBJECTIVE: To describe associations between anemia and hookworm (Necator americanus) infection in hospitalized women in rural Chiapas, Mexico. MATERIAL AND METHODS: We retrospectively reviewed the hospital records of 68 anemic women (defined as having a hemoglobin level < 10 mg/dl) or with intestinal parasitism, identified through hospital discharge and transfusion records for the calendar year 1999. The medical charts of 86 cases were located, 18 of which were not confirmed as anemia cases. The hospital is located in Altamirano, Chiapas. Characteristics of subjects were compared using Student's t-test (for continuous variables) and the chi 2 test (for categorical variables). A p-value < or = 0.01 was used for statistical significance. Chart review and data analysis took place during the year 2000. RESULTS: Fifty percent of women who had stool examinations were infected with N. americanus. Necator often coexisted with other potential causes of anemia, such as pregnancy and hemorrhage. Hemoglobin levels in hookworm infected women (mean 4.1 g/dl) were significantly lower than in uninfected women (mean 7.0 gm/dl), and Necator prevalence was significantly higher in the anemic women (50%) than in the overall hospital population (1.9%). CONCLUSIONS: Anemic women should be offered stool testing where Necator is present, and should be considered for antihelminthic treatment even if pregnant. Further investigation is recommended among women in Chiapas, and probably elsewhere in Mexico. The English version of this paper is available too at: http://www.insp.mx/salud/index.html. PMID- 12736989 TI - Knowledge and beliefs about malaria transmission and practices for vector control in southern Mexico. AB - OBJECTIVE: To investigate the knowledge and beliefs about malaria transmission and practices for vector control in eight villages on the coastal plain of Chiapas, Mexico. MATERIAL AND METHODS: A cross-sectional survey was conducted during May and June 1995 in Chiapas, Mexico. A questionnaire to investigate family structure, knowledge on malaria transmission, preventive measures and attitudes towards seeking treatment was applied to both family heads of a sample of households. Associations were analyzed by estimating odds ratios with confidence intervals and p values, using bivariate and multivariate logistic regression methods. RESULTS: Malaria knowledge was poor and only 48% associated malaria with mosquito bites. The perceived benefit of indoor residual spraying was associated to a reduction of mosquitoes, a reduction in the numbers of cockroaches and rats, but only 3% associated it directly with the prevention of malaria transmission. Most villagers (97.6%) agreed with the indoor residual spraying of insecticides. Ninety nine percent of villagers had mosquito bednets, 75.7% used them all year round. Other measures used by villagers to prevent mosquito bites were smoke and mosquito coils. Above 40% of villagers self medicated when any member of the family had a fever episode, but 51% attended proper health services (community dispensary, private physician, health worker). About 61% used pesticides for agricultural or livestock purposes and 55% applied themselves. Women had a greater participation as family health promoters, with 70% of the housewives being in charge of the application of self-protection preventive measures. CONCLUSIONS: Educational programs aimed at increasing awareness on the participation of mosquitoes on malaria transmission could promote community participation in malaria control in the region. The English version of this paper is available too at: http://www.insp.mx/salud/index.html. PMID- 12736991 TI - Contact with hospital syringes containing body fluids. Implications for medical waste management regulation. AB - OBJECTIVE: To determine amount of syringes used in the hospital and extent of contact with blood and body fluids of these syringes. MATERIAL AND METHODS: Syringe use was surveyed at a tertiary care center for one week; syringes were classified into the following four categories according to use: a) contained blood; b) contained other body fluids (urine, gastric secretion, cerebrospinal fluid, wound drainage); c) used exclusively for drug dilution and application in plastic intravenous (i.v.) tubes, and d) for intramuscular (i.m.), subcutaneous (s.c.), or intradermic (ID) injections. RESULTS: A total of 7,157 plastic disposable syringes was used; 1,227 (17%) contained blood during use, 346 (4.8%), other body fluids, 5,257 (73%) were used exclusively for drug dilution and application in plastic i.v. lines, and 327 (4.5%) were utilized for i.m., s.c., or ID injections. An estimated 369 140 syringes used annually, or eight syringes per patient per in-hospital day. All syringes were disposed of as regulated medical waste, in observance of the law. CONCLUSIONS: There is an urgent need to review recommendations for medical waste management by both international agencies and local governments, based on scientific data and a cost-benefit analysis, to prevent resource waste and further environmental damage. The English version of this paper is available too at: http://www.insp.mx/salud/index.html. PMID- 12736993 TI - [Basic information on National Health System services, 2001]. PMID- 12736994 TI - [Agreement on international tobacco control]. PMID- 12736992 TI - Control of Chagas disease vectors. AB - Most Latin American countries are making dramatic progress in controlling Chagas disease, through a series of national and international initiatives focusing on elimination of domestic populations of Triatominae, improved screening of blood donors, and clinical support and treatment of persons infected with Trypanosoma cruzi. Some countries, particularly Uruguay, Chile and Brazil, are sufficiently advanced in their programmes to initiate detailed planning of the subsequent phases of Chagas disease control, while others such as Peru, Ecuador, and Mexico, are currently applying only the initial phases of the control campaigns. In this review, we seek to provide a brief history of the campaigns as a basis for discussion of future interventions. Our aim is to relate operational needs to the underlying biological aspects that have made Chagas disease so serious in Latin America but have also revealed the epidemiological vulnerability of this disease. The English version of this paper is available too at: http://www.insp.mx/salud/index.html. PMID- 12736995 TI - [SARS: a new public health challenge]. PMID- 12736997 TI - Renal anatomy and overview of nephron function. AB - The kidneys are highly vascularized organs that are responsible for maintaining the body's internal environment. The nephron is the basic structural and functional unit of the kidney involved in the complex interplay between tubular and vascular components resulting in the excretion of fluid and solutes. The regulation of body fluid and solutes is governed by the processes of filtration, reabsorption, and secretion. Unwanted fluid and substances are excreted from the body as urine. PMID- 12736996 TI - Staff nurses' perceptions of the work environment in freestanding hemodialysis facilities. AB - While one suggested cause of the current nursing shortage is nurses' negative perceptions of the work environment, little is known of nephrology nurses' perceptions of the dialysis work environment. The purpose of this study was to assess the extent to which staff nurses who work in freestanding hemodialysis facilities rate the presence of organizational characteristics common to magnet hospitals in their current job. Study findings indicate that staff nurses in hemodialysis units identify several notable features of magnet hospitals in their work settings. However, a majority of nurses disagreed that many attributes of magnet hospitals are present in hemodialysis work environments. This study provides a preliminary description of some of the factors that affect nurses' perceptions of the work environment in freestanding dialysis facilities. Further work is needed in this area. PMID- 12736998 TI - Expanding the living organ donor pool: positive crossmatch and ABO incompatible renal transplantation. AB - There is a significant shortage of donor kidneys. As a result, kidney transplant candidates wait for prolonged periods of time for an organ, and over eight die every day while awaiting a kidney transplant. To improve this situation, the transplant community has actively sought creative solutions to the organ shortage. Many patients have willing live donors who are excluded from donation due to a positive crossmatch or blood group incompatibility. Plasmapheresis and intravenous immunoglobulin in combination can efficiently remove antibodies against donor tissue and blood group antigens and prevent these antibodies from returning after transplantation. Both strategies are complex but have good success rates and provide an opportunity for transplantation to those who might wait years for an organ or die waiting on the list Knowledge of these novel protocols is essential for the nephrology nurse who is often the first health care provider to discuss transplantation with end stage renal disease (ESRD) patients. PMID- 12736999 TI - Why are hemodialysis treatments shortened and skipped? Development of a taxonomy and relationship to patient subgroups. AB - Shortening and skipping hemodialysis treatments occur commonly and are associated with inadequate dialysis and increased mortality. These behaviors are also frequently equated with patient noncompliance. The prevalence and demographic correlates of shortening and skipping treatments have been studied extensively. However, less is known about the reasons for shortening and skipping treatments and whether patient characteristics correlate with specific reasons. Understanding the relationship between reasons and patient characteristics is essential for determining whether patient subgroups are at a disadvantage in achieving optimal dialysis and may help guide future interventions. Semistructured interviews were conducted with 168 selected patients who shortened or skipped treatments. A variety of reasons were responsible for shortening and skipping hemodialysis treatments. Content analysis of patient responses revealed five categories of reasons for shortening and skipping, including medical problems, technical problems, life tasks, transportation, and patient decisions. The most common reasons for shortening were medical problems (38%) and life tasks (24%), while the most common reasons for skipping were life tasks (33%) and transportation (22%). Furthermore, patient subgroups differed in the reasons for shortening and skipping. After multivariate adjustment for patient characteristics, technical problems were more common among women. Life tasks were more common among men, younger patients, and patients with hypertension. Transportation problems were more common among African Americans. Interventions to optimize hemodialysis treatment should identify and target patient-specific reasons for shortening and skipping and not assume noncompliance. PMID- 12737000 TI - Addressing patient concerns about intravenous iron therapy. AB - Hemodialysis patients may not fully understand the relationship between anemia and kidney disease and the role of intravenous (i.v.) iron therapy in anemia management. To address this, nephrology nurses have willingly undertaken the role of patient educator in addition to technical expert. This responsibility is challenging because nurses need to understand the rationale for i.v. iron therapy and the efficacy and safety characteristics of the available i.v. iron therapies, which include iron dextran, iron sucrose, and sodium ferric gluconate. Nurses must then be able to effectively communicate this knowledge to their patients in an understandable fashion. This manuscript offers suggestions to nurses for approaches to address patient concerns about i.v. iron therapies, the latest information regarding the efficacy and safety of i.v. iron treatments, and strategies to communicate this information to patients. PMID- 12737002 TI - Dry weight: to be set or not to be ... that is a good question. AB - The dialysis procedure should be individually tailored to the client at all times. Learn each client's signs and symptoms for a hypotensive occurrence as well as other signals that suggest the need to adjust fluid removal when aiming for or establishing a new dry weight. Clients may display a variety of indicators, and it is imperative to familiarize yourself as much as possible with client-specific symptoms. Some clients also exhibit specific symptoms prior to actually experiencing a blood pressure drop. Learning these "red flags" is also important. PMID- 12737001 TI - Relationships between hemoglobin and hematocrit levels and hospitalization and survival rates in dialysis patients. AB - Preserving life and reducing or limiting morbidity are the primary goals of dialysis therapy and the focus of day-to-day nursing assessments and interventions. A series of clinical analyses have shown that partial correction of anemia (to maintain hemoglobin [Hb] in the range of 11 to 12 g/dL and hematocrit [Hct] levels in the range of 33% to 36%) is associated with a 10% to 74% decrease in the risk of death and a 7% to 58% decrease in the risk of hospitalization compared with lower Hb (Hct) levels. These clinical data illustrate the importance of maintaining all dialysis patients at Hb levels > or = 11 g/dL and Hct levels > or = 33%. More recently, a series of publications have suggested that additional incremental improvements in mortality and morbidity rates may be realized when Hb and Hct levels are maintained above the target range recommended by the NKF-K/DOQI. While these results are intriguing, the safety and efficacy of this approach require further study and clarification. PMID- 12737003 TI - Peritoneal dialysis fact sheet. PMID- 12737004 TI - HIPAA: how dialysis providers will be affected. PMID- 12737005 TI - The role of the general practitioner and the dietitian in patient nutrition management. AB - There is mounting evidence that nutrition plays an important role in the aetiology and management of many diseases affecting Australians. Nutrition counselling provides a strategy for not only reducing patient suffering, but also for reducing the health care costs associated with these illnesses. At the forefront of providing nutrition counselling to Australians are General Practitioners (GPs) and Dietitians. Australian data shows that GPs encounter many patients with the chronic diseases that have nutrition in their aetiology and management. Although this presents opportunities to provide nutrition counselling, overseas literature suggests that often nutrition counselling opportunities are not taken up. At present, there is little evidence to support whether this problem exists in Australia, or the magnitude of the problem. Whilst evidence suggests there are barriers for GPs in providing nutrition counselling, there is conflicting evidence on how these influence the GP's decision to provide such counselling. Overseas studies have also identified barriers for GPs to refer to dietitians to provide nutrition counselling, however there is no evidence to identify whether these barriers exist in Australia. Whilst various strategies have been implemented to aid in the provision of effective nutrition management to the Australian public, there is little evidence on the efficacy of these. Research is needed in the above areas if effective patient nutrition management is to be implemented in Australia. PMID- 12737006 TI - Bioactive phytochemicals in Indian foods and their potential in health promotion and disease prevention. AB - Besides being a source of nutrients, foods, particularly plant foods, are a rich source of bioactive phytochemicals or bionutrients. Studies carried out during the past 2-3 decades have shown that these phytochemicals have an important role in preventing chronic diseases like cancer, diabetes, coronary heart disease and hyper-cholesterolaemia. The major classes of phytochemicals with disease preventing functions are dietary fibre, antioxidants, detoxifying agents, immunity-potentiating agents and neuropharmacological agents. Each class of these functional agents consists of a wide range of chemicals with differing potency. For example, antioxidant function is exhibited by some nutrients, such as vitamin E, vitamin C and provitamin A. Other phytochemicals that have antioxidant properties are carotenoids, phenolic compounds, flavonoids and isothiocyanates. Some of these phytochemicals have more than one function. Foods rich in these chemicals and exhibiting disease-protecting potential are called functional foods. Indian habitual diets, which are based predominantly on plant foods like cereals, pulses, oils and spices, are all good sources of these classes of phytochemicals, particularly dietary fibre, vitamin E, carotenoids and phenolic compounds. There is, however, much scope for further systematic research in screening Indian foods and diets for these phytochemicals and assessing their potential in protecting against chronic diseases. PMID- 12737007 TI - Commodities consumed in Italy, Greece and other Mediterranean countries compared with Australia in 1960s and 1990s. AB - Consumption figures for 15 major commodities (cereals, wheat, rice, maize, potato, pulses, olive oil, other vegetable oils, vegetables, fruits, wine, meats, animal fats, milk + products, and fish + seafood) were collected from FAO Food Balance Sheets during the 1960s (1961-1969) and late 1990s (1995-1999). For some nutritionists the "model Mediterranean diet" is the Italian or Greek diet of the 1960s, for others the concept of Mediterranean countries is more general. Analysis shows: (1) In the 1960s, Australia consumed more meat, milk, animal fat than Italy or Greece and less cereals, wheat, pulses, olive oil, vegetables, fruits and wine. (2) In the 1960s, Australia's olive oil, vegetables, fruits and wine consumption were within the range for all 18 Mediterranean countries (i.e. Spain, France, Italy, Malta, Croatia, Bosnia, Albania, Greece, Cyprus, Turkey, Syria, Lebanon, Israel, Egypt, Libya, Tunisia, Algeria and Morocco). (3) In the 1990s, food consumptions have evolved; Australia's wine and milk consumption is now similar to Italy and Greece; consumption of wheat, olive oil, vegetables, fruits and fish are lower; consumption of potato, pulses, other vegetable oils and meat are higher than Italy or Greece. (4) Australia's consumption of the 15 commodities is within the range of all Mediterranean countries in the late 1990s, except wheat consumption was lower. PMID- 12737008 TI - Documenting the reasons people have for choosing their food. AB - Understanding the reasons that people have for choosing their food, and why these choices vary, may affect the dietary advice and assumptions about the nutrient adequacy of future food intake. One group of respondents living in Jakarta, Indonesia completed two interviews with the same combined food frequency and qualitative technique, called Food Choice Map (FCM) over a one-month period. Another group of Indonesian respondents from a town in Java completed an FCM interview and a 24-hour recall interview. The Food Choice Map identified the same major foods as contributing to individual intakes as are identified by a 24-hr recall interview. The FCM also identified reasons for changes in food choice. The reasons for food choices varied less than the different food items chosen. The FCM links data on dietary behaviours with perceptions that respondents use to explain of those behaviours. Such data can be used to develop communication strategies for health promotion. PMID- 12737009 TI - Pacific pandanus fruit: an ethnographic approach to understanding an overlooked source of provitamin A carotenoids. AB - Commonly recommended plant sources of provitamin A, such as dark green leafy vegetables, are not acceptable in many population groups. The objective of this study was to identify other indigenous foods that may be effectively promoted to alleviate vitamin A deficiency (VAD) and to gather information relevant to identification, production, acquisition, and consumption of foods relevant to a food-based VAD prevention strategy in the Federated States of Micronesia. An ethnographic study on edible pandanus cultivars, involving key informant interviews and observation was carried out. Analyses revealed a great range in carotenoid content. Several orange-coloured pandanus cultivars, all highly acceptable, contained high levels of carotenoid, almost meeting daily requirements in usual consumption patterns, whereas light yellow-coloured cultivars contained low levels. Availability has decreased substantially in recent years due to increased consumption of imported foods and general neglect of indigenous foods. High-carotenoid pandanus should be promoted for general enjoyment and health benefits. PMID- 12737010 TI - Effect of maturity on macromineral content of selected leafy vegetables. AB - Macro mineral contents were estimated in commonly consumed green leafy vegetables in India, namely; Koyyathotakura and Peddathotakura (varieties of Amaranthus species); Erragogu and Tellagogu (variety of Hibiscus species); Gangabayalakura (Portulaca olereceo) and Palak (Spineces olerecea) at three different stages of maturity. Varietal differences were also observed. The results of the study showed that as the plant matured from stage I (15 days) to stage II (30 days) calcium and magnesium content increased. In contrast, phosphorus content decreased as the plant matured. Varietal differences were also observed at different stages of maturity. The results also indicated that the consumption of green leafy vegetables at stage I (15 days) and stage II (30 days) potentially provide the greatest amount of minerals. PMID- 12737011 TI - Fatty acid and sterol composition of frozen and freeze-dried New Zealand Green Lipped Mussel (Perna canaliculus) from three sites in New Zealand. AB - In view of previously reported anti-inflammatory bioactivity of the New Zealand Green Lipped Mussel (NZGLM), the overall lipid profile and fatty acid and sterol composition of the NZGLM from various sites in New Zealand (Hallam Cove, Port Ligar. Little Nikau) were investigated using thin layer chromatography (TLC) and gas liquid chromatography (GLC). Samples were either frozen (F) or freeze-dried (FD) soon after collection. It was also thought prior to the study, there may be differences in the dietary sources of phytoplankton between the sites, responsible for the bioactivity, however data collected in New Zealand reported no difference in the type of phytoplankton, but a difference in the quantity. There were no major significant differences in the major components of the lipid, fatty acid and sterol composition between FD or frozen samples, nor were there any significant differences in the major composition between sites. The only major difference was between total lipid composition of the freeze-dried and frozen samples due to the removal of water during freeze-drying. Total lipid content on a dry weight basis in FD samples was 8.4 g/100 g tissue and was significantly higher than frozen samples (P < 0.05) and there was no significant site variation. The lipid class content between sites was also not significantly different as judged by TLC. Triglyceride (TG) lipid fraction appeared to be the most prominent in the frozen and FD samples. The free fatty acid (FFA) band was the next most prominent band and was visually more prominent in the frozen samples. Sterol esters (SE) were detected in higher amounts in the frozen samples compared with the FD samples. Phospholipid (PL) and sterols (ST) were distributed throughout all samples. Polyunsaturated fatty acids (PUFA) were the main group of fatty acids in both FD and frozen samples (45-46%), most of which were omega-3 (n 3) fatty acids (40-41%). Saturated fatty acids (SFA) accounted for approximately one quarter of total fatty acids, with little variation between FD and frozen samples. The major fatty acids of the NZGLM were docosahexaenoic acid (DHA: 22:6 n-3) (19% in both FD and frozen samples), eicosapentaenoic acid (EPA; 20:5 n-3) and palmitic acid (16:0) (15% in both FD and frozen samples). Cholesterol was the most prominent sterol (31% of total sterols). Other major sterols included desmosterol/brassicasterol (co-eluting), 24-methylenecholesterol, trans-22 dehydrocholesterol, 24-nordehydrocholesterol and occelasterol. This study is unique as it compares the lipid composition of the NZGLM from three sites in New Zealand with the additional effect of processing. This is the second comparative study investigating the lipid, fatty acid and sterol composition of the NZGLM with added interest in the effect of freeze-drying on the lipid content of the mussel. This study showed that there were no major significant differences in lipid, sterol and fatty acid composition between the FD and frozen samples of the NZGLM for three sites in New Zealand. Food chain studies and further research is warranted to investigate the presence and role of major and minor lipid components of the NZGLM. PMID- 12737012 TI - Effect of nutrition support on immunity in paediatric patients with beta thalassaemia major. AB - Nutritional deficiencies have been variably observed in thalassaemia and the aetiology of many of the immune abnormalities in thalassaemic children are poorly defined. Therefore, we tested the hypothesis that certain immune abnormalities have a nutritional basis. Nutritional status, selective quantitative and functional indices of immunity were studied in twelve children (7 females, 5 males; mean age 28 months, SD 5 and range 19.8-35.5), with thalassaemia major before and after a one month period of intensive nutrition support (the study diet consisted of 'Enfapro' liquid formula (Mead Johnson) with added dextrose and corn oil to achieve a caloric density of 1.1 kcal/cc in addition to vitamins and minerals). Each child was provided approximately 150 kcal/day and 4 g of protein/day. Lymphocyte proliferation to Concanavalin A (Con A) (P = 0.008) and Purified Protein Derivative (PPD) (P = 0.002) was depressed upon entry into the study, however the response to Con A attained normal values by the end of the intervention. Compared to baselines, the proliferative response to Con A (P = 0.005) and Phytohemagglutinin A (PHA) (P = 0.031) both improved after the nutrition support. Although there was no general correlation of zinc status with lymphocyte proliferation, normal baseline zinc status was associated with improvement of proliferation. The %CD4 increased (P = 0.036), primarily because of a decrease in total lymphocytes and to lesser extent a decrease in CD8 lymphocytes. Serum immunoglobulin concentrations were found to be elevated on admission but were not significantly affected by the nutrition intervention. C3 concentrations were uniformly depressed on admission but increased by the end of the study protocol (P = 0.037). C4 and CH50 activity were not significantly influenced by the intervention. In conclusion, children with beta thalassaemia have abnormalities of lymphocyte function as well as key complement components that are responsive to nutrition support. In addition, zinc status appears to have an important role in lymphocyte function in these children. PMID- 12737013 TI - The prevalence of childhood obesity in primary school children in urban Khon Kaen, northeast Thailand. AB - Childhood obesity is a serious public health problem because of its strong association with adulthood obesity and the related adverse health consequences. The published literature indicates a rising prevalence of childhood obesity in both developed and developing countries. However no data exists on the prevalence in Northeast Thailand, one of the poorest regions of the country and one that has experienced a recent economic transition. The objective of this study was to estimate the prevalence of obesity in seven to nine year old children in urban Khon Kaen, Northeast Thailand. A cross-sectional school based survey was conducted to determine the prevalence of obesity in children of urban Khon Kaen, Thailand. Multi-staged cluster sampling was used to select 12 school clusters of 72 children each between the ages of 7 and 9 years, in primary school grades 1, 2 and 3 from government, private and demonstration schools. A total of 864 seven to nine year old school children were studied. Anthropometric measurements of standing height and weight were taken for all subjects to the nearest tenth of a centimetre and tenth of a kilogram respectively, Childhood obesity was defined as a weight-for-height Z-score above 2.0 standard deviations of the National Center for Health Statistics/World Health Organisation reference population median. The prevalence of childhood obesity was 10.8% (95% CI: 7.6, 13.9). Obesity was significantly more prevalent in boys than girls. The biggest difference was observed between the three school types, with the highest prevalence of obesity found at teacher training demonstration schools and the lowest at the government schools. This study provides the first data on childhood obesity prevalence in Northeast Thailand. The prevalence of 10.8 per cent is lower than that found in two other urban areas of Thailand but slightly higher than expected for this relatively poor region. If this prevalence rate increases, as observed in other countries in economic transition, the incidence of non-communicable diseases associated with obesity is also likely to increase, thus raising cause for concern and reason for intervention to both control and prevent obesity during childhood. PMID- 12737014 TI - The relationships between anthropometric measurements, serum vitamin A and E concentrations and lipid profiles in overweight and obese subjects. AB - The weight, height, body mass index (BMI), waist/hip ratio, serum retinol and alpha-tocopherol and lipid profiles of 16 overweight (BMI > or = 25.0 kg/m2) Thai males and 56 overweight females, compared with 14 males and 58 females in a control group (BMI 18.5-24.9 kg/m2), were investigated. Subjects for the study were those persons who turned up regularly for physical check-up at the Outpatient Department, General Practice Section of Rajvithi Hospital, Bangkok. The study was conducted between December 2000-March 2001. Higher levels of cholesterol, LDL-C, LDL-C/HDL-C ratio were found in the overweight compared with the control subjects. Statistically significant higher triglyceride levels were found in the overweight compared with the control subjects. The median serum retinol concentration in overweight subjects was 2.80 mumol/L (range 0.53-4.62 mumol/L) compared with 2.97 mumol/L (range 1.21-4.12 mumol/L) in control subjects (p = 0.0736). The median serum alpha-tocopherol concentration in overweight subjects was 17.30 mumol/L (range 6.29-28.65 mumol/L) compared with 18.75 mumol/L (range 5.30-30.28 mumol/L) in control subjects (P < 0.05). The median values of retinol and alpha-tocopherol serum concentrations in the overweight and obese males were lower than those of the overweight and obese females. A total of 6.3% (1 out of 16) and 12.5% (2 out of 16) of the overweight/obese males had decreased retinol and alpha-tocopherol levels, while the overweight/obese females had decreased retinol and alpha-tocopherol level of 1.8% (1 out of 56) and 10.7% (6 out of 56), respectively. A total of 12.5% and 39.3% of the overweight/obese males and females had cholesterol concentrations of > or = 6.48 mmol/l. However, the prevalence of low HDL-C (HDL-C < or = 0.91 mmol/l) was found to be 50% in the overweight and obese males and 10.7% in the overweight and obese females. Statistically significant associations were found between age, cholesterol, LDL C, and serum alpha-tocopherol in the overweight and obese male and female subjects. A negative correlation was found between weight, BMI, AC, MAMC, hip circumference and serum retinol in both the overweight and obese subjects. A negative correlation was found between weight, BMI, MAMC, waist, hip circumferences and serum alpha-tocopherol in both the overweight and obese subjects. PMID- 12737015 TI - Predictive equations for estimation of stature in Malaysian elderly people. AB - Height is an important clinical indicator to derive body mass index (BMI), creatinine height index and also to estimate basal energy expenditure, basal metabolic rate and vital capacity through lung function. However, height measurement in the elderly may impose some difficulties and the reliability is doubtful. Equations estimating height from other anthropometric measures have been developed for Caucasians, but only one study has developed an equation (based on arm span only) for an Asian population. Therefore, a cross sectional study was conducted to develop equations using several anthropometric measurements for estimating stature in Malaysian elderly. A total of 100 adults (aged 30 to 49 y) and 100 elderly subjects (aged 60 to 86 y) from three major ethnic groups of Malays (52%), Chinese (38.5%) and Indians (9.5%) participated in this study. Anthropometric measurements included body weight, height, arm span, half arm span, demi span and knee height were carried out by trained nutritionists. Inter and intra observer errors and also % Coefficient Variation (%CV) were calculated for each anthropometric measurement. Equations to estimate stature were developed from the anthropometric measurements of arm span, demi span and knee height of adults using linear regression analysis according to sex. Elderly subjects were shorter and lighter compared to their younger counterparts. The %CV of anthropometric measurements in adults and elderly subjects ranged between 5 to 6%, with standing height having the lowest %CV. When the equations derived from adults were applied to elderly subjects, it was found that percentage difference between actual height and the estimated value ranged from 1.0 to 3.3%. However, the percentage difference between estimated height from the equations developed in this study compared to those derived from the equations of other populations ranged between 0.2 to 8.7%. In conclusion, standing height is an ideal technique for estimating the stature of individuals. However, in cases where its measurement is not possible or reliable, such as in elderly subjects, height can be estimated from proxy indicators of stature. In this study arm span showed the highest correlation with standing height, which is in agreement with other studies. It should be borne in mind that equations derived from taller statured populations (e.g. Caucasians) may be less accurate when applied to shorter statured populations. PMID- 12737016 TI - Vitamin B12 and folate status of older New Zealand women. AB - The primary aim of this study was to assess the biochemical vitamin B12 and folate status of a representative group of elderly women (70-80 y) living in Dunedin, New Zealand. A second aim was to determine the prevalence of hyperhomocysteinaemia and to explore the determinants of homocysteine (hcy) concentration in this population. A cross-sectional study was carried out between June and August of 2000. Two hundred and fifty women were randomly selected from the 1998 electoral roll. Fasting blood samples were analysed for folate, vitamin B12, total hcy, creatinine, and haematological parameters. Of the women selected, 87 did not respond, 37 were not traceable, 23 were not eligible or had died, and 103 agreed to participate. The overall response rate was 46%. Based on a cut-off of 150 pmol/L for serum B12, 13% of participants would be classified as having sub-optimal vitamin B12 status. Of the women, 3 and 5%, respectively, had low serum (< 6.6 nmol/L) and erythrocyte folate (< 317 nmol/L) concentrations. No participant had megaloblastic anaemia. The prevalence of hyperhomocysteinaemia (> 15 mumol/L) in this population was 18%. Hyperhomocysteinaemia in this group may be partly explained by renal insufficiency because there was a significant association between serum creatinine and plasma hcy (P < 0.001). Blood folate levels but not serum B12 were significantly inversely associated with hcy. In conclusion, there was a moderately high prevalence of hyperhomocysteinaemia and suboptimal plasma vitamin B12 concentrations but not low blood folate concentrations in this elderly female population. PMID- 12737017 TI - Homocysteine concentrations lowered following dietary intervention in an aboriginal community. AB - Low circulating folate concentrations lead to elevations of plasma homocysteine. Even mild elevations of plasma homocysteine are associated with significantly increased risk of cardiovascular disease (CVD). Available evidence suggests that poor nutrition contributes to excessive premature CVD mortality in Australian Aboriginal people. The aim of the present study was to examine the effect of a nutrition intervention program conducted in an Aboriginal community on plasma homocysteine concentrations in a community-based cohort. From 1989, a health and nutrition project was developed, implemented and evaluated with the people of a remote Aboriginal community. Plasma homocysteine concentrations were measured in a community-based cohort of 14 men and 21 women screened at baseline, 6 months and 12 months. From baseline to 6 months there was a fall in mean plasma homocysteine of over 2|mol/L (P = 0.006) but no further change thereafter (P = 0.433). These changes were associated with a significant increase in red cell folate concentration from baseline to 6 months (P < 0.001) and a further increase from 6 to 12 months (P < 0.001). In multiple regression analysis, change in homocysteine concentration from baseline to 6 months was predicted by change in red cell folate (P = 0.002) and baseline homocysteine (P < 0.001) concentrations, but not by age, gender or baseline red cell folate concentration. We conclude that modest improvements in dietary quality among populations with poor nutrition (and limited disposable income) can lead to reductions in CVD risk. PMID- 12737018 TI - Current status of vitamin A deficiency and the National Vitamin A Control Program in Nepal: results of the 1998 National Micronutrient Status Survey. AB - The overall objective of the Nepal Micronutrient Status Survey (NMSS) was to assess the distribution and severity of micronutrient malnutrition, and to measure the progress achieved by different interventions. Data presented in this paper concern the prevalence of vitamin A deficiency (VAD) and the outreach and coverage of the National Vitamin A Supplementation activity. A multi-stage cluster sample design was employed that provided statistically representative data for each of thirteen eco-development strata (because of low population density, the West Mountains, Mid-west Mountains and Far-west Mountains were combined into a single stratum). The design allowed for aggregate estimates to be made at the national and ecological zone level. The survey showed a significant improvement in the status of clinical vitamin A deficiency in Nepal. The prevalence of both Bitot's spots and night-blindness among preschool children decreased from levels observed in surveys conducted in the previous twenty years. However, the prevalence of night-blindness was found to be 5% among women, and over 1% among school-aged children, which indicates that the entire population is vulnerable to VAD. These observations support findings from other surveys that have noted a high prevalence of maternal night-blindness in Nepal. Biochemical data collected as part of the survey indicated a high prevalence of low serum retinol (< 0.70 mumol/l), particularly among preschool children. Almost one of every three children (32.3%) and one of every six women (16.6%) had low serum retinol values. Low serum retinol among preschool children was associated with young age (6-11 months), rural location, wasting, presence of night-blindness and Bitot's spots, and residence in the Terai or Mountains. Similarly, sub-clinical VAD in women was associated with age (less than 20 years), pregnancy, the presence of night-blindness and residence in the Terai or Mountains. In the 42 districts covered by the National Vitamin A Programme (NVAP), more than 87% of preschool children were reached with vitamin A capsules. In addition to this, the National Immunisation Day (NID) provided oral polio vaccine drops to an estimated 95.7% of children 12-59 months. Awareness of the importance of vitamin A was, however, much higher in the NVAP districts than in non-programme districts. As would be expected, clinical VAD was most prevalent among children who had not received vitamin A during the most recent vitamin A capsule distribution. Indeed, the data show that vitamin A capsule receipt among children conferred a 59% protective effect for night-blindness and a 51% effect for Bitot's spots. These results point to significant progress having been achieved by the NVAP and NID capsule distribution activities. PMID- 12737019 TI - Early infant feeding practices in Jinan City, Shandong Province, China. AB - To investigate early infant feeding practices in Jinan, China, a cross-sectional study was carried out in April-May 2000. Data were collected through structured interviews among nearly all the mother-infant pairs (247) in four communities in the city with infants under seven months of age. All but one was born in a hospital and rooming-in (infant sleeping in same room as mother) was widely practiced. Breastfeeding was practiced universally, but first contact with the new-born was delayed one or more hours for 51% of subjects. Colostrum was given to 94% of the infants, yet during the first three days, all but 34% were given water, artificial baby milk, glucose or other prelacteal feeds. Among infants under 4 months of age, 35% were currently exclusively breastfeeding, but at two months only 40% were, and only half that many had received nothing but breast milk since birth. Exclusive breastfeeding has probably increased in China, but further promotion is still needed in this area. PMID- 12737021 TI - Exercise-trained but not untrained rats maintain free carnitine reserves during acute exercise. AB - Exercise training is known to induce physiological adaptations that improve exercise performance and alter patterns of energy substrate utilization to favour fatty acid oxidation. L-Carnitine is an essential cofactor for the oxidation of fatty acids under all physiological conditions, including exercise. This study evaluated the effect of acute exercise on carnitine concentrations in tissue and serum, liver carnitine palmitoyltransferase-I activity and expression, and serum lipids in both trained and untrained rats as compared to non-exercised rats. Serum acyl- and total carnitine was significantly higher in the trained animals, whether exercised or not, suggesting an exercise-induced increase in a renal threshold for carnitine. Untrained rats had significantly higher acylcarnitine in skeletal muscle and an acyl/free carnitine ratio of 0.63 +/- 0.06 compared with 0.31 +/- 0.16 in trained animals receiving an identical acute bout of exercise, demonstrating that untrained animals utilized a significantly higher percentage of free carnitine reserves during exercise. This study suggests that free carnitine reserves may be reduced during exercise in untrained rats, an effect that has the potential to impair both carbohydrate and fat metabolism during exercise. PMID- 12737022 TI - Creating new conversations. PMID- 12737020 TI - Postprandial metabolic responses to milk enriched with milk calcium are different from responses to milk enriched with calcium carbonate. AB - The purpose of this study was to assess whether there are any differences in postprandial physiological responses to skim milk powder enriched with milk calcium (SMP + milk calcium) and skim milk powder enriched with calcium carbonate (SMP + CaCO3), with each of the milks providing 1200 mg calcium. This was a randomised, controlled, crossover study involving 16 men and 29 women over 55 years of age. Measurements of calcium and bone metabolism were taken after an overnight fast before each drink, and postprandially every hour for 8 h. The impact of time and drink on the responses was analysed by repeated measures of analysis of variance. Serum calcium was significantly higher between 2 and 8 h after consumption of SMP + CaCO3 compared with SMP + milk calcium (P < 0.0001). Serum phosphate was significantly higher between 2 and 5 h after drinking the SMP + milk calcium compared with SMP + CaCO3 (P < 0.0001). The level of parathyroid hormone (PTH) was virtually unchanged after consumption of SMP + milk calcium, but decreased between 1 and 4 h after SMP + CaCO3 (P = 0.02). The serum C telopeptide level, a marker of bone resorption, was significantly lower after SMP + CaCO3, compared with SMP + milk calcium, between 4 and 8 h after drinking the milk (P < 0.05). We conclude that serum calcium levels have a higher increase after SMP + CaCO3 consumption than after SMP + milk calcium consumption, and that this is associated with lower serum PTH concentrations and a more prolonged postprandial decrease in bone resorption. PMID- 12737023 TI - The Romanow Commission: an opportunity lost. PMID- 12737024 TI - You are what you serve: healthy and environmentally friendly food service. PMID- 12737025 TI - A public preference for home care. PMID- 12737027 TI - Reflections on Romanow. PMID- 12737026 TI - Chronic complications of diabetes: cardiovascular and kidney disease. PMID- 12737028 TI - After Kirby and Romanow: where to from here? PMID- 12737029 TI - Where do we go from here? AB - In the past two and a half years since the Senate Committee began its study of the Canadian healthcare system, an abundance of reports on healthcare, at both the provincial and federal levels, have been issued that have provided the public and policy-makers with a comprehensive overview of the key issues and the main options for change. We have clearly reached a point where action of some sort can no longer be deferred on the grounds that further study is necessary. It is now a question of finding the political will to move forward with reform. PMID- 12737030 TI - Issues in the governance of Canadian hospitals, Part II: Hospital planning. AB - A hospital's board is directly accountable to the corporation of the hospital. Together they are accountable to the patients and communities served by the hospital, to the provincial government that funds the hospital on behalf of these patients and communities, and to the staff of the hospital. Thus, the board of trustees has a critical role in defining the purposes, principles and objectives of hospitals. PMID- 12737031 TI - Are longer hospital stays beneficial for the elderly? AB - Utilization review is a way to manage healthcare costs and is widespread in Canada, as managers attempt to use available acute-care beds in a best practice manner. As we reduce beds and decrease length of stay, we often wonder if the outcomes for patients are affected, particularly if the patients are elderly. PMID- 12737032 TI - Cultivating rural healthcare professionals: the Norfolk General Hospital Health Science Perspectives Program. AB - Studies around the world have shown that where healthcare professionals establish their practices is influenced by where they grow up and receive their training. In our Simcoe, Ontario-based Norfolk General Hospital (NGH), about 40% of our physicians and 80% of other medical health professionals study nearby and return here to their rural roots. PMID- 12737033 TI - House of healing, house of disrespect: a Kantian perspective on disrespectful behaviour among hospital workers. AB - Respect toward patients is one of the most fundamental and central tenets guiding both modern bioethical practice and the everyday behaviour of all healthcare professionals. However, similar courtesy and respect is often breached in day-to day interactions between hospital workers. Many examples are relatively minor, while egregious examples such as gender discrimination and physical abuse do occur. The more egregious transgressions may be handled by formal processes, even legal proceedings. However, the innumerable smaller examples of disrespectful behaviour are ubiquitous and insidious in their erosion of a productive collaborative approach to patient care and other aspects of functioning within the institution. The authors briefly summarize some of the pertinent literature on this subject and analyze the problem of disrespect in the hospital with special focus on the issue as seen from the perspective of the moral philosophy of Immanuel Kant. Some simple recommendations for improving disrespectful behaviour amongst hospital workers are offered. PMID- 12737034 TI - Healthcare, race and diversity: time to act. AB - Nearly 40 years after the enactment of civil rights legislation in the United States and following a generation of multicultural policy in Canada, the vast majority of nurses in both countries are still female and white. But while nursing remains starkly segregated by gender, it is hardly alone among the health professions in its under-representation of visible minorities. Physicians and other caregivers find themselves in similar positions--they do not always reflect the communities and patients they serve. PMID- 12737035 TI - Telephone contact centre links to primary healthcare reform. AB - A new telephone health advice service implemented in September 2000 is integral to the Capital Health (Edmonton) Region's strategy to build a comprehensive approach to primary healthcare. Capital Health Link operates 24 hours a day, seven days a week and was the first comprehensive nurse-based health advice phone line in Alberta. PMID- 12737036 TI - Aramark profile. Interview by Cynthia Martin. PMID- 12737037 TI - Adverse drug reactions. PMID- 12737039 TI - Perspectives. Competitive Medicare has merits, won't save much, analysts say. PMID- 12737038 TI - The doctor is out: physician participation in the Rationed Access Day work stoppage in British Columbia, 1998/99. AB - BACKGROUND: In 1998/99, the British Columbia Medical Association (BCMA) asked physicians to withdraw elective services on a series of 20 Rationed Access Days (RADs). This work stoppage was called to protest continued free proration triggered by total physician billings exceeding a fixed budget cap. This paper examines how physicians' practice activity changed on RADs, the rates of participation in this job action and characteristics of those physicians who participated. DESIGN: Population-based cohort study of physicians based on administrative data. POPULATION: All full-time physicians billing the provincial healthcare plan. METHODS: Participation in the RAD initiative was inferred by comparing physicians' average daily billings on RADs, Sundays, holidays and regular weekdays. Using thresholds established from these distributions, the number of RADs observed by each physician in 1998/99 was calculated and examined in relation to their demographics, location and prior years of practice activity. RESULTS: For the 4,131 physicians studied, average daily payments on RADs were similar to those on Sundays and holidays but much lower than those on non holiday, non-RAD weekdays. Using billing thresholds of $200 (for GPs) and $400 (for specialists), we found a high degree of participation with the study population observing a median of 15 of the 20 scheduled RADs. While there were some differences in participation among age groups, geography and the prior years of practice activity, the differences were small. INTERPRETATION: This study found high solidarity in the BCMA's 1998/99 RAD initiative. Most full-time fee for-service FS physicians appeared to participate in at least three-quarters of the 20 scheduled RADs. PMID- 12737040 TI - Integron content of Salmonella enterica serotype Typhimurium strains isolated in Hungary in the years 1997-1999. AB - The integron content of 52 DT104/U302 phage type strains and 53 non-DT104/U302 strains of Salmonella enterica serotype Typhimurium (S. Typhimurium) was studied in PCR experiments using a 5'-CS/3'-CS primer pair (Levesque et al., 1995). Forty three out of 44 streptomycin- and/or ampicillin-resistant DT104 and related phage type strains were found to carry a 1 kb and/or 1.2 kb long integron. The other resistance markers did not affect the number and size of integrons; no integron free multidrug-resistant (MDR) DT104 strains were found. The two large groups of DT104 strains (Felix-Callow's phage types 2 and 2c) proved to be identical in respect of integron patterns (IPs), supporting the views of those authors who consider DT104 a single clone. Strains of human and animal origin did not differ from each other in their IPs. Within the non-DT104 phage types, ampicillin- and/or streptomycin-resistant, integron-free MDR strains were also found. Based on amplicons varying between 290 and 3500 bp an IP system was suggested. The commonest amplicon sizes in non-DT104 strains were 1450 and 2050 bp. The IPs of DT104 strains and of non-DT104 strains containing an integron of 1 and 1.2 kb size were stable. In contrast, the IPs of other non-DT104 strains showed a varying degree of instability. Integron loss was frequently associated with spontaneous plasmid elimination and changes of R-type among the descendants of a given strain. PMID- 12737041 TI - Analysis of gene cassettes of streptomycin-spectinomycin resistance of Hungarian Salmonella enterica serotype Typhimurium strains. AB - By PCR using the ant(3")-Ia primer pair the aadA gene was detected in 34 streptomycin- and spectinomycin-resistant Salmonella enterica serotype Typhimurium strains. Out of them 12 belonged to DT104 and 22 to non-DT104 phage type. Using different primer combinations it was demonstrated that this gene was integron-associated in all cases: in the DT104 strains it was generally contained by a 1 kb integron while in the majority of the non-DT104 strains by a 2.05 kb (less often by a 1.9 or 1 kb) integron. In the case of integrons carrying multiple cassettes the cassette containing the aadA gene was located closer to the 3' end of the integron. The aadA genes of DT104 and non-DT104 strains were different: in the former group the aadA2 gene, while in the latter group (constituted by strains of five different phages types as well as unclassifiable and untypable strains) the aadA1 gene could be identified. The RH50/RH51 primer pair described by Collis and Hall (1992) proved to be suitable for rapid discrimination between the aadA1 and aadA2 genes on the basis that the RH51 primer bound exclusively to the aadA2 gene. PMID- 12737042 TI - Comparison of methods for the diagnosis of equine herpesvirus type 1 infection. AB - The objective of the investigations was to study the occurrence of the equine herpesvirus type 1 (EHV-1) infection in aborted equine fetuses and in newborn foals and to compare the sensitivity of virus isolation, immunohistochemistry and histology in 101 cases and of fetal serology in 68 cases in the diagnosis of the infection. Out of the 93 aborted equine fetuses and 8 weak foals, 15 (14.9%) (14 fetuses and 1 foal) proved to be EHV-1 infected by immunohistochemical and 13 (12.9%) by virological investigation. Characteristic microscopic changes were seen in several organs in all cases, while intranuclear inclusion bodies could be found only in 25 (35.2%) of the 71 virus-positive tissue samples. Four (5.9%) cases proved to be positive by fetal serological investigation, but none of these cases showed any EHV-1 specific lesions and in none of these cases could the virus be detected by virus isolation or by immunohistochemistry. According to the results, fetal serology does not seem to be a useful test in virus-positive cases, while the immunohistochemical method seems to be a reliable and a slightly more sensitive method than virus isolation in the diagnosis of EHV-1 infection. PMID- 12737043 TI - Seroprevalence of Lawsonia intracellularis in large pig production units. AB - In 11 'farrow-to-finish' outdoor or indoor production units, blood samples from late pregnant gilts were tested by indirect immunofluorescence antibody (IFA) serum assay for Lawsonia intracellularis. The offspring of positively tested gilts were tested at 2, 7, 12, 17, 22 and 27 weeks of age for seroprevalence of Lawsonia intracellularis. All offspring of IFA positive gilts were seronegative at 2 and 7 weeks of age. At 12 weeks of age 81.0% of indoor and 51.0% of outdoor pigs were tested positive. While at 17 weeks of age 82.5% of indoor-raised pigs showed seropositivity, in outdoor units the seropositivity declined to 31.3%. At weeks 22 and 27 indoor-raised pigs still showed marked seropositivity (17.7% and 11.5%) but their outdoor-raised counterparts revealed declining values (7.4% and 0%). PMID- 12737044 TI - Effect of dietary fumonisin B1 on certain immune parameters of weaned pigs. AB - Only few data are available on the effect of fumonisins on the immune response. The aim of the present study was to examine whether dietary fumonisin B1 (FB1) has any effect on the humoral and cellular immune response in weaned pigs, depending on the dose and the time of toxin exposure. Fusarium moniliforme fungal culture was added to the experimental animals' diet to ensure an FB1 intake of 1, 5 and 10 ppm (first experiment) or 100 mg per animal per day (second experiment). The control animals were fed a toxin-free diet. In order to determine the immune response, the animals were vaccinated against Aujeszky's disease with inactivated vaccine (Aujesping K, Phylaxia-Sanofi, Budapest, Hungary). Specific and nonspecific in vitro cellular immune response was measured by the lymphocyte stimulation test (LST) induced by PHA-P, Con A, LPS and inactivated suspension of the Aujeszky's disease virus. Humoral immune response, e.g. specific antibody titre, was measured by the virus neutralisation (VN) test. None of the immunological parameters examined showed significant differences between groups. It could be concluded that fumonisin B1 had no significant effect on the humoral and cellular specific and nonspecific immune response when fed in a high dose (100 mg/animal/day for 8 days) or in a low concentration even for a longer period (1, 5 and 10 ppm for 3-4 months). PMID- 12737045 TI - Prevalence of hydatidosis and fertility of hydatid cysts in sheep in Kirikkale, Turkey. AB - This study was conducted in order to determine the prevalence and fertility of hydatid cysts in sheep in Kirikkale, Turkey. A total of 3.2% of 553 lambs and 50.9% of 1320 adults were found to be infected with hydatid cysts. The most common locations of cysts were in the liver and lung. Liver cysts tended to be more fertile than lung cysts (81.53% v. 76.47%), though the difference was not significant. The mean number of viable protoscoleces in the liver and lung cysts were 12,400 and 5,800, respectively. In general, 1-10 cysts were found in liver (51.8%) and lung (64.7%). The findings of this study indicate that hydatid cysts are common in sheep and the fertility rate of cysts is quite high. Sheep play an important role in the life cycle of Echinococcus granulosus in this region. PMID- 12737046 TI - Disappearance of ethanol from isolated sheep rumen. AB - The absorption of ethanol from the rumen was studied in three British Milk sheep equipped with a rumen cannula. After removal of the rumen content and washing the forestomachs several times the reticulo-omasal orifice was closed and through the cannula 20 or 60 ml ethanol and 2 ml Cr-EDTA were infused in physiological saline. The entire fluid volume was 3000 ml. At the start of the experiment (0 min) and subsequently in the 5th, 15th, 30th, 45th, 60th and 75th minutes samples were taken from the fluid present in the forestomachs. During the 75-min experiment the amount of ethanol gradually decreased in the rumen. The rate of disappearance varied according to concentration. The graph depicting the change of ruminal ethanol concentration shows a curve typical of passive transport. The equation describing the disappearance of ethanol was y = -0.0474x2 + 5.6544x + 10.869 after the administration of 20 ml ethanol, and y = -0.1377x2 + 19.541x - 24.606 after the infusion of 60 ml ethanol. It was established that ethanol was absorbed through the rumen wall by a passive transport process. PMID- 12737047 TI - Growth hormone and insulin-like growth factor-I action on progesterone secretion by porcine corpora lutea isolated at various periods of the luteal phase. AB - This study was conducted to investigate the interactions between growth hormone (GH) and insulin-like growth factor-I (IGF-I) on progesterone (P4) secretion by porcine luteal cells cultured in vitro. Cells isolated from corpora lutea (CL) collected at three different periods of the luteal phase (CL1--early luteal phase; CL2--middle luteal phase and CL3--late luteal phase) were incubated with different doses of GH (10, 100 or 200 ng/ml). After 48 h cultures were terminated and the media were frozen until further P4 concentration analysis. GH (100 ng/ml) increased P4 secretion by CL1 and CL2 and had no effect on CL3. In separate studies these cells were treated for 48 h with IGF-I alone or with GH combined with IGF-I. IGF-I alone increased basal P4 secretion only by cells collected from CL1 while concurrent treatment with GH had no effect on P4 secretion by any type of CL. To investigate the possible mechanism of GH and IGF-I mediated induction of P4 secretion, an inhibitory study was conducted. In this experiment, luteal cells collected from CL1 were cultured in the absence or presence of cycloheximide (an inhibitor of protein synthesis) or actinomycin D (an inhibitor of DNA transcription). Cycloheximide or actinomycin D completely blocked the stimulatory effect of both GH and IGF-I on P4 production but did not reduce basal progesterone secretion suggesting involvement of gene transcription and translation in the GH and IGF-I action on luteal cells. Additionally, the activity of 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) under the influence of GH added alone or together with IGF was measured by the conversion of pregnenolone to progesterone. Stimulation of P4 secretion in P5-treated cells in GH-stimulated cultures was not observed, however, high stimulatory effect was noted in IGF-I treated cultures. In conclusion, the present studies indicate that there is direct and cycle stage dependent influence of GH and IGF-I on steroidogenesis in procine luteal cells. It is suggested that both IGF and GH may exert some regulatory action during CL development in the pig. PMID- 12737048 TI - Effect of the El Nino phenomenon on the ovarian responsiveness and embryo production of donor cows. AB - The effects of different Temperature Humidity Index (THI) values in cold, hot and El Nino (EN) climates on superovulation and embryo production were analysed on Holstein Friesian donor cows. There were significant differences in the THI among the three climates. The average temperature in the EN period was 6 degrees C higher than in the summer period of the previous 30 years. The number of corpora lutea (CL) and embryos were log- and back-transformed, Kolmogorov-Smirnoff test was used for normality and Lilliefors test was applied for significance. In the cold season THI was 70.74 +/- 1.35 and the average number of CL was 9.84 +/- 4.37. In the hot season the THI was 73.99 +/- 0.72 and the average number of CL was 9.70 +/- 4.49. When the THI, in the EN period, increased up to 79.74 +/- 4.01, the superovulation response was significantly (P < 0.01) reduced (average number of CL = 5.22 +/- 2.53). The embryo production result showed a similar tendency. In the hot period the average number of embryos obtained was 5.87 +/- 2.98. However, in the EN period it decreased to 4.21 +/- 2.05. Higher temperature reduced embryo quality. The proportion of live embryos (%) was 59.2 +/- 37.4 in the cold and 38.2 +/- 38.5 in the EN periods of the year (P < 0.01). However, ovarian sensitiveness showed adaptation to summer environment while the heat stress, which was more severe in the EN period, negatively affected the superovulation response and embryo production. PMID- 12737050 TI - A survey for BVDV antibodies in cattle farms in Slovakia and genetic typing of BVDV isolates from imported animals. AB - A serological survey for bovine viral diarrhoea virus (BVDV) antibodies on a collection of 1295 serum samples obtained from 6-12 months old cattle originating from 45 farms in Slovakia was carried out. On 13 farms more than 90% of the examined animals were seropositive, on 14 farms 71-90% seroprevalence was observed, on 13 farms only 50-70% animals were found to be positive for BVDV antibodies, while the remaining 5 farms showed fewer than 50% seropositive animals. The average incidence of BVDV antibodies (around 70%) was similar as determined 30 years ago. Of 84 serum samples from seronegative animals originating from 14 farms in which 70-98% seropositivity was observed, six were positive in Ag-BVDV ELISA indicating persistently infected (PI) cattle. On a farm to which animals were imported from abroad, a BVD outbreak was observed. Of 110 animals tested, four were positive in Ag-ELISA indicating the presence of PI cattle on this farm. Genetic typing of two isolates from imported animals performed by RT-PCR (324/326 primers from 5'-UTR), sequencing of PCR products and computer-assisted phylogenetic analysis revealed that they belong to BVDV-1 h group. PMID- 12737049 TI - Effect of ovariohysterectomy on canine postsurgical leukocyte function. AB - The effect of surgery on phagocytic activity of blood leukocytes and mitogen induced blastogenesis of lymphocytes was studied in fourteen dogs. Simple ovariohysterectomy with anaesthesia induced by ketamine and xylazine or by ketamine, xylazine and halothane caused a short nonsignificant depression of phagocytic activity that persisted for four hours after surgery. Ingestion capacity of leukocytes decreased significantly immediately after surgery. Mitogen induced blastogenesis of lymphocytes was depressed significantly in the first 48 hours and despite partial recovery this parameter did not reach the value of the control groups until the end of observation (7 days). A more conspicuous decrease of blastogenic response of blood lymphocytes to mitogens was found after the use of ketamine and xylazine in a dose maintaining anaesthesia. Anaesthesia with ketamine and xylazine in the lower dose and maintained with halothane resulted in a later improvement of the blastogenic response of lymphocytes. PMID- 12737051 TI - Epitopic characterisation of Turkish bovine viral diarrhoea virus (BVDV) isolates using monoclonal antibodies. AB - In this study, 15 bovine viral diarrhoea viruses (BVDV) isolated from the field in Turkey were characterised for their biotype, cloned and eventually analysed for their epitopic composition in terms of glycoprotein E2. Immunoplaque assay, plaque assay, limiting dilution and streptavidin-biotin-peroxidase techniques were used for biotype characterisation, cloning of cytopathic (cp) and noncytopathic (ncp) biotypes and epitope analysis, respectively. While 14 out of 15 BVDV isolates were distinguished as ncp biotype, 1 isolate was found to be containing both biotypes (cp + ncp). According to the reactivity patterns of isolates with 15 monoclonal antibodies, 4 different antigenic groups could be formed. There were no antigenic differences between the isolates derived from the same animal with various time intervals. On the other hand, biotype clones isolated from the same animal exhibited difference in one epitope. This is the first study describing antigenic characterisation of BVDV field isolates in Turkey. PMID- 12737053 TI - Effects of a cycled morphological intervention on selected suppletive BE forms. AB - The purpose of this study was to demonstrate the efficacy of a cycled morphological intervention involving copula is, by comparison to a control group, and to examine the effectiveness of this intervention in producing change in untreated auxiliary is and am. Twenty-two preschool children, who produced copula is but no other suppletive forms of BE as targets, received six sessions of naturalistic intervention cycled over 24 weeks. Production accuracy levels for treated copula and untreated auxiliary is and am were obtained from spontaneous language samples elicited after 12 and 24 weeks of intervention. The cycled intervention resulted in significant improvement in copula is in comparison to the no-treatment control group at the 12 week sampling point; similar gains were observed for the untreated auxiliary is, but not am. After 24 weeks, however, gains in untreated am were significantly greater than for copula is, providing new data to suggest that generalization may also be observed for the suppletive am. PMID- 12737052 TI - Planning of syllables in children with developmental apraxia of speech. AB - The aim of the present study was to investigate whether children with developmental apraxia of speech (DAS) show a deficit in planning syllables in speech production. Six children with DAS and six normally speaking (NS) children produced high- and low-frequency of occurrence syllable utterances, in which the syllable structure was systematically manipulated in an otherwise unchanging phoneme sequence. Anticipatory coarticulation, using second formant trajectories, and durational structure were analysed. The results showed stronger coarticulation in the children with DAS when compared to the normally speaking children. but in contrast to our expectations, in neither group was a systematic effect of syllable structure on the second format trajectory found. Effects of syllable structure did emerge for durational structure in that durational adjustments were found in the segments of the second syllable. These adjustments were less systematic in children with DAS when compared to normally speaking children. Furthermore, at the prosodic level, normally speaking children showed metrical contrasts that were not realized by the children with DAS. The latter results are interpreted as evidence for a problem in the planning of syllables in speech production of children with DAS, in particular concerning prosodic aspects, which is discussed in relation to the automation of speech production. PMID- 12737054 TI - Variation in articulatory timing of three English consonants: an electropalatographic investigation. AB - Increasingly, electropalatography (EPG) is being used in speech pathology research to identify and describe speech disorders of neurological origin. However, limited data currently exists that describes normal articulatory segment timing and the degree of variability exhibited by normal speakers when assessed with EPG. Therefore, the purpose of the current investigation was to use the Reading EPG3 system to quantify segmental timing values and examine articulatory timing variability for three English consonants. Ten normal subjects repeated ten repetitions of CV words containing the target consonants /t/, /l/, and /s/ while wearing an artificial palate. The target consonants were followed by the /i/ vowel and were contained in the carrier phrase 'I saw a _.' Mean duration of the approach, closure/constriction, and release phases of consonant articulation were calculated. In addition, inter-subject articulatory timing variability was investigated using descriptive graphs and intra-subject articulatory timing variability was investigated using a coefficient of variation. Results revealed the existence of inter-subject variability for mean segment timing values. This could be attributed to individual differences in the suprasegmental features of speech and individual differences in oral cavity size and structure. No significant differences were reported for degree of intra-subject variability between the three sounds for these same phases of articulation. However, when this data set was collapsed, results revealed that the closure/constriction phase of consonant articulation exhibited significantly less intra-subject variability than both the approach and release phases. The stabilization of the tongue against the fixed structure of the hard palate during the closure phase of articulation may have reduced the levels of intra-subject variability. PMID- 12737055 TI - High-level language difficulties in Parkinson's disease. AB - Twenty-six subjects with idiopathic Parkinson's disease (PD) and normal cognitive status (as measured by the Mini-Mental State Examination) were examined with a battery of tests selected to reveal subtle and/or high-level language impairments. The test battery included 'repetition of long sentences', 'recreating sentences', 'making inferences', 'comprehension of logico-grammatical sentences', 'comprehension of ambiguous sentences' and 'comprehension of metaphors', 'word definitions', 'word fluency', 'naming', 'sentence analysis' and 'morphological completion'. Comparisons were made between the PD subjects and 26 control subjects matched for age, gender and level of education. Significant differences in performance between the PD subjects and the control subjects were found in the ability to make inferences and to analyse sentences (state the correct number of words in a read sentence). An additional four subjects with different degrees of cognitive dysfunction were also investigated and were found to have particular problems in making inferences, recreating sentences and comprehending metaphors and ambiguities. The results suggest that processing implied information might be a specific problem in this group and that the task of making inferences could be a particularly sensitive test of high-level language dysfunction. PMID- 12737056 TI - [Give credit where credit is done]. PMID- 12737057 TI - [Norwegian program for malignant lymphomas]. PMID- 12737058 TI - [After seventy all is trouble and sorrow]. PMID- 12737059 TI - [Compulsory admissions in Norwegian psychiatry]. PMID- 12737060 TI - [Compulsory admissions for observation in emergency psychiatric departments- discharge next day?]. PMID- 12737061 TI - [Compulsory medical treatment in an emergency psychiatric department]. PMID- 12737062 TI - [Sensory-motor interaction and development of muscle pain]. PMID- 12737063 TI - [Occurrence of depression in the elderly--a systematic review of 55 prevalence studies from 1990-2001]. PMID- 12737064 TI - [Apoptosis and cardiovascular disease]. PMID- 12737065 TI - [The significance of written routines for care after crib death and child accidents]. PMID- 12737066 TI - [Natural course of breast cancer and effect of mammographic screening]. PMID- 12737067 TI - [Surgical treatment of urolithiasis]. PMID- 12737068 TI - [Prevention of respiratory syncytial virus infections with palivizumab]. PMID- 12737069 TI - [Use of MR examination in knee injuries]. PMID- 12737070 TI - [Hand allotransplantation]. PMID- 12737071 TI - [Parkinsonism and use of selective serotonin uptake inhibitors]. PMID- 12737072 TI - [The physician as advocate--an attempt at an analysis of the Perruche case]. PMID- 12737073 TI - [Discovery of DNA--the project, the effort and the prestige]. PMID- 12737074 TI - [How are prices for important drugs stipulated in poor countries?]. PMID- 12737075 TI - [Johan Bremer--an unusual scientist in Norwegian psychiatry]. PMID- 12737076 TI - [How many physicians do we need?]. PMID- 12737077 TI - [Without proportions about drugs]. PMID- 12737079 TI - [Disability and patient economy]. PMID- 12737078 TI - [Introduction and evaluation of screening]. PMID- 12737080 TI - [Cancer--the inner enemy]. PMID- 12737081 TI - [Mysteries in publishing houses and text]. PMID- 12737082 TI - [Study of the social representations of drugs and cannabis: motivation for prevention]. AB - Drugs and cannabis are two complex, multifaceted social objects. Two exploratory research projects were conducted from semi-directive interviews (n = 50) and questionnaires (n = 300) in order to investigate the social representations contained in these two objects according to both the social characteristics of individuals (gender, age) and their proximity to cannabis (consumption versus non consumption). The study of their representations allows for emphasis to be placed upon the socio-cognitive reconstruction that individuals maneuver, reconstruction largely influenced by the social insertion of individuals but also by their method of cannabis consumption. Knowledge of these representations seems to be a tool for understanding the often complex realities linked to the world of drugs and allows for reflection upon more adapted prevention methods. PMID- 12737084 TI - [Prevalence of obesity amount elementary school children in Grenoble]. AB - Facing the growing increase of childhood obesity in France, the Grenoble Municipal School Health Service wanted to evaluate the importance of this phenomenon present among elementary school children in the area. A descriptive, cross-sectional and retrospective survey was conducted during the 2000/2001 academic year among a group of 1361 children born in 1990. The Quetelet Index was utilized to measure the level of being overweight, and the breaking point levels for being overweight and obese were defined according to new international standards. The prevalence of obesity is approximately 3% (2.7% in girls and 2.9% in boys) and remains constant from nursery school to elementary school. Nearly 35% of obese children in nursery school remain obese at the end of elementary school. Overweight prevalence is at its highest and increases significantly from nursery school (8.3%) through elementary school (14%). The study further revealed that girls aged 10-12 who did not pass and were held back from the next grade are five times more at risk of obesity than those who pass. The Municipal School Health Service has a critical role to play in the prevention of childhood obesity and its early screening and detection. PMID- 12737083 TI - [Satisfaction of patients hospitalized in Ho Chi Minh City, Vietnam]. AB - This study aims to describe the level of satisfaction of patients and their families, according to the type of hospital and the sociodemographic characteristics of patients hospitalised within Ho Chi Minh City (HCMC), Vietnam. The study is supported by a sample of 538 patients and their families from all of the hospitals in HCMC to whom an evaluation questionnaire was given to be filled out on the day of their release. The average age is 39, and 64.4% are women. The scores measuring the level of satisfaction vary from 57.7 to 90.7 points (on a scale of 0 to 100). The section under the heading "treatment provided by the doctors" received the highest scores (90.7/100). Dissatisfaction was primarily associated with factors such as the amount of time spent waiting, the behaviour of the hospital staff, cleanliness, the cafeteria and parking for motorbikes. Almost 100% of the patients spoke about their experiences during their stay in the hospital, and 50.2% had some negative opinions (1.042 complaints registered). The older patients, having a lower level of education and living either in other cities or in the rural region of HCMC, tented to be more satisfied than the younger patients, possessing higher levels of education and generally residing in the sub-urban or urban regions of HCMC. The patients who judged their condition to be very serious and their problems bad enough to justify hospitalisation, and who also felt that their health and condition had improved at the time of release compared to the time of admission were more satisfied than the other patients. Patients hospitalised who had an individual private room in a specialised hospital tended to be more satisfied that those who had a room sharing many beds for several patients in either a general hospital or in the emergency area of a specialised hospital. The authors observed no correlation between the level of satisfaction and the patient's gender. The results of this study have highlighed that certain domains, notably hat of the hospital's environment, should be reviewed and examined by health care administrators and managers in order to ensure the quality of the patient's care and coverage. PMID- 12737085 TI - [Attitudes regarding the delivery of formal and informal care: comparison of French and Chilean adolescents]. AB - The representations that youth have of health professionals and young people's demands in terms of the operation and administration of services create an original and complex problematic. Clearly, this originality implies the important differences from one culture to another. For this very reason, it seemed that a comparative study relating the representations and attitudes confronted when care is sought by young people from countries with different cultural contexts would assist in comprehending why adolescents have such particular ways of using--or not using--formal and self-administered health services. An original open-ended response questionnaire was jointly designed and validated by a French and Chilean team. A mutually agreed upon sample of 957 school children, adolescents aged from 14 to 19, participated in the study in France and in Chili. The following correlations were found. In the event of a sleeping problem (or other general worry that is physically manifested), the mother is the privileged confidant, and in the specific case of a relationship or emotional problem, it is usually one of the adolescents' friends. The general practitioner is the favoured professional person in the event of a purely physical problem. When confronted with an emotional problem, one-third of adolescents say that they would not consider going to a consultation. The expectations of the French toward health professionals are more often within the "emotional" arena than those of the Chileans which generally concern the "medical/technical" field. The practice of self-administered care is qualitatively similar but the French prefer taking medication whereas the Chileans prefer the "little home remedies". The use of natural medicine is more widespread among young Chileans, but the types of medicine used are similar, namely herbal teas and other plant-based remedies and homeopathy. These results have a variety of implications, especially in terms of the need for training health professionals in the consideration of emotional and relationship problems. It is desirable that the official health care sector considers the care delivered outside of it as being complementary resources, which respond to the adolescents' need for autonomy, and then integrate those contributions into its own area of financial responsibility. PMID- 12737086 TI - [Role of patient associations in the development of therapeutic education in France]. AB - The aim of this survey conducted among patient associations is to define the role and the position that they have with regard to the development of therapeutic education in France. 124 associations were solicited (out of over 500 existing), and 68 replied. 17 indicated that the survey was not relevant for them. 51 answered the 43 questions related to the conception and implementation of educational programmes, their goals and objectives, the choice and selection of educational activities, the target audiences and pedagogical principles of reference. The results obtained demonstrate the predominance of informational activities and psychological support. The responding associations declared that sometimes they conduct educational activities which rather resemble informational activities. Only three associations declared having Implemented and managed formalised educational programmes based on pedagogical methods. The aims and objectives most frequently targeted were focused on increasing the patients' knowledge on their disease and its treatment. These educational programmes are usually delivered by members of the association's office staff. However, overall most of the responding associations indicated that it is relatively difficult to provide precise data on the pedagogical methods of the activities undertaken. In light of the results, it is therefore necessary to consider the totality of the activities conducted by the associations as a mechanism for building educational resources in which the place of formalised educational programmes remains marginal, even quasi-absent, for the moment. Associations believe that patient education is an important issue for their development. It is highly likely that the emergence of such programmes will only be possible if the associations show some degree of autonomy in relation to the health care sector and assert a point of view specific to patients, above and beyond that of health care users. PMID- 12737087 TI - [School health education: practices and representations of primary school teachers]. AB - A better understanding of primary school teachers' practices and representations toward health education (HE), and identification of individual or structural resistance as well as the partnership and training needs all constitute important goals in HE research. A quantitative study was conducted between April and December 2001 on a representative sample of the population of primary school teachers (n = 673) in the Auvergne region. The results demonstrate that the majority of teachers declare practicing and implementing HE. The approach is primarily thematic, essentially limited to a few lessons since it is only integrated into a broader project 20% of the time. 30% of teachers work in partnership, mainly with partners in school health; however, parents are rarely involved in HE activities. Parameters which influence the teachers' practices and representations are: (1) prioritizing work within an educational network and an inter-communal pedagogical regrouping, with the advantage that there are a greater number of teachers to do HE in these schools than in others, and (2) receiving initial or in-service training. These results suggest that a policy aiming to generalize the inclusion of HE in French primary schools must develop teacher training as well as support and accompany the collective dynamics within schools. PMID- 12737088 TI - [Globalization and destruction of frameworks and conditions for life]. PMID- 12737089 TI - [The five-year public health law: the SFSP's analysis and recommendations]. PMID- 12737090 TI - [Medical perspective in the case of a hunger strike. The problem of providing treatment under constraints]. PMID- 12737091 TI - Secular changes in tooth size and dental arch dimensions in the mixed dentition. AB - Secular changes in the mixed dentition were studied. Permanent tooth size and dental arch dimensions were examined in Norwegian children born in the 1960s and 1980s, Swedish children born in the 1960s and 1980s, Norwegian Sami children born in the 1980s, and a sample of Norwegian skulls dating from the 14th to the 19th century. The Norwegian Sami children were nomadic in the summertime. A sample of pigs was studied before and after a maceration process to determine what dimensional changes might occur in such a process. A shrinkage of 0.3%-1.7% was found. This information was used when the skulls were compared with the modern groups. Lateral dental arch lengths were shorter in the children born in the 1960s compared with the children born in the 1980s. This was a result of the higher prevalence of caries in the second deciduous molars in the 1960s groups. Children who had lost a deciduous canine prematurely were found to have smaller dental arch perimeters. When compared with other data, this was blamed on a pre existing crowding. Permanent tooth size was smaller in the skulls compared with the modern groups. Improved nutrition is considered to be the main reason for the difference. Relative dental arch space differed in the group born in the 1960s from that in the other groups, indicating a greater prevalence of crowding in the former. Relative dental arch space in the skulls and in the group born in the 1980s was similar. A more traditional way of living, as practised by the Sami group in this thesis, was not favorable for relative dental arch space. The transverse intermaxillary relation in boys changed from the 1960s to the 1980s, which indicated that the 1980s group ran a greater risk of developing a posterior cross-bite. Before the same conclusion could be made in the girls, the mesial drift of the first permanent molars had to be corrected for, because of a higher prevalence of caries in the 1960s group. The sex-pooled analysis of the skulls and the contemporary groups revealed that the risk for developing a posterior cross-bite in the 1980s group was greater than in the skulls. The skulls had smaller arch depths than the modern groups. PMID- 12737092 TI - Neurofeedback treatment for attention-deficit/hyperactivity disorder in children: a comparison with methylphenidate. AB - Clinical trials have suggested that neurofeedback may be efficient in treating attention-deficit/hyperactivity disorder (ADHD). We compared the effects of a 3 month electroencephalographic feedback program providing reinforcement contingent on the production of cortical sensorimotor rhythm (12-15 Hz) and betal activity (15-18 Hz) with stimulant medication. Participants were N = 34 children aged 8-12 years, 22 of which were assigned to the neurofeedback group and 12 to the methylphenidate group according to their parents' preference. Both neurofeedback and methylphenidate were associated with improvements on all subscales of the Test of Variables of Attention, and on the speed and accuracy measures of the d2 Attention Endurance Test. Furthermore, behaviors related to the disorder were rated as significantly reduced in both groups by both teachers and parents on the IOWA-Conners Behavior Rating Scale. These findings suggest that neurofeedback was efficient in improving some of the behavioral concomitants of ADHD in children whose parents favored a nonpharmacological treatment. PMID- 12737093 TI - The effects of specific respiratory rates on heart rate and heart rate variability. AB - In this study respiratory rates of 3, 4, 6, 8, 10, 12, and 14 breaths per minute were employed to investigate the effects of these rates on heart rate variability (HRV). Data were collected 16 times at each respiratory rate on 3 female volunteers, and 12 times on 2 female volunteers. Although mean heart rates did not differ among these respiratory rates, respiratory-induced trough heart rates at 4 and 6 breaths per minute were significantly lower than those at 14 breaths per minute. Slower respiratory rates usually produced higher amplitudes of HRV than did faster respiratory rates. However, the highest amplitudes were at 4 breaths per minute. HRV amplitude decreased at 3 breaths per minute. The results are interpreted as reflecting the possible effects of the slow rate of acetylcholine metabolism and the effect of negative resonance at 3 cycles per minute. PMID- 12737094 TI - Effects of daily activities on ambulatory blood pressure during menstrual cycle in normotensive women. AB - The purpose of this study was to determine whether (1) there is a menstrual phase effect on blood pressure (BP) and heart rate (HR), and (2) the effects of physical effort, posture, or moods on BP and HR is mediated by the menstrual phase. Twelve normotensive women, aged between 28 and 50, with normal menstrual cycles were studied. BP was measured at 30- to 60-min intervals during a 24-hr period using an ambulatory BP monitor on Days 1, 8, 15, and 22 of the menstrual cycle. Participants were asked to report their posture, physical effort, and mood ("annoyed," "tense," and "happy") on 5-point Likert-type scales each time the ambulatory BP monitor took measurements. Systolic BP (SBP) was lower on Day 8 of the cycle. Diastolic BP (DBP) and HR were lower on Days 1 and 8. Daytime SBP was affected by the time of the day and posture, but not by moods, whereas daytime DBP was affected by posture and levels of tenseness. The level of physical effort only affected HR, not BP. The average daytime physical and emotional variables had little influence over the average daytime BP. In 12 normotensive women with a normal menstrual cycle, SBP was lower during the follicular phase and DBP and HR were lower during the follicular phase and menstruation even after controlling the effects of other factors. Physical activity or moods had only momentary effects on BP or HR. A cross-validation statistical method used is suggested to study how individuals are affected by various factors. With the use of this method, the inclusion of menstrual phase in the model improved the prediction of SBP for 5 out of the 12 women studied. PMID- 12737095 TI - Biofeedback of R-wave-to-pulse interval normalizes blood pressure. AB - We investigated whether biofeedback of the R-wave-to-pulse interval, a measure related to the pulse wave velocity, enables participants with either high or low arterial blood pressure to modify their blood pressure. Twelve participants with high blood pressure (mean systolic blood pressure = 142.6 +/- 13.5 mmHg; mean diastolic blood pressure = 99.9 +/- 12.3 mmHg) and 10 participants with low blood pressure (mean systolic blood pressure = 104.8 +/- 6.6 mmHg; mean diastolic blood pressure = 73.2 +/- 4.2 mmHg) received 3 individual sessions of RPI biofeedback within a 2-week period. Participants with high blood pressure were rewarded for decreasing and participants with low blood pressure for increasing their blood pressure. Standard arm-cuff blood pressure measurements across the sessions served as dependent variables. Participants with high blood pressure achieved significant reductions of systolic (15.3 mmHg) and diastolic (17.8 mmHg) blood pressure levels from the beginning of the first to the end of the last training session. In contrast, participants with low blood pressure achieved significant increases in systolic (12.3 mmHg) and diastolic (8.4 mmHg) blood pressure levels. The degree of blood pressure changes in this study might be of clinical relevance. With prolonged and refined training regimens, even larger effects seem to be likely. PMID- 12737096 TI - Biofeedback therapy in the colon and rectal practice. AB - In coloproctology, biofeedback has been used for more than 20 years to treat patients with fecal incontinence, constipation, and rectal pain. It can be performed in a number of conditions with minimal risk and discomfort. However, it does require the presence of some degree of sphincter contraction and rectal sensitivity. Biofeedback can be time-consuming and demands motivation. The purpose of this paper is to review the indications, methodology, and results of anorectal biofeedback in the treatment of these disorders. Mean success rates for biofeedback range from 72.3% for fecal incontinence of diverse etiology, 68.5% for constipation attributable to paradoxical puborectalis syndrome, and 41.2% for idiopathic rectal pain. However, criteria to define success vary tremendously among researchers and there is a tendency to indicate biofeedback in a myriad of conditions when other therapeutic options, including surgery, fail or are inappropriate. These factors make comparison of the results difficult and reinforce the need for randomized controlled trials and studies assessing long term follow-up. In summary, biofeedback is a simple, cost-effective, and morbidity-free technique and remains an attractive option, especially considering the complexity of the functional disorders of the colon, rectum, anus, and pelvic floor. PMID- 12737097 TI - Nutrition in the treatment of attention-deficit hyperactivity disorder: a neglected but important aspect. AB - Attention-deficit hyperactivity disorder (ADHD) is multidetermined and complex, requiring a multifaceted treatment approach. Nutritional management is one aspect that has been relatively neglected to date. Nutritional factors such as food additives, refined sugars, food sensitivities/allergies, and fatty acid deficiencies have all been linked to ADHD. There is increasing evidence that many children with behavioral problems are sensitive to one or more food components that can negatively impact their behavior. Individual response is an important factor for determining the proper approach in treating children with ADHD. In general, diet modification plays a major role in the management of ADHD and should be considered as part of the treatment protocol. PMID- 12737098 TI - Building relationships that inspire service. PMID- 12737099 TI - Retaining staff the magnet way: fostering a culture of professional excellence. PMID- 12737100 TI - Exposing the myths of employee satisfaction. PMID- 12737101 TI - Improving medication safety. Suppliers share ways to build momentum for medication safety initiatives. PMID- 12737102 TI - Addressing bioterrorism. What ethical issues and questions surround potential responses to bioterrorist attacks? PMID- 12737103 TI - Job hunting on the Web. One key to an effective job search is knowing how and where to look. PMID- 12737104 TI - Developing a regional referral strategy. Regional strategies can build physician relationships and expand your customer base. PMID- 12737105 TI - Creating a mentoring culture. Learn steps for establishing a formal mentoring system in your organization. PMID- 12737106 TI - Competing for talent. Combating the labor shortage requires a mix of short- and long-term strategies. PMID- 12737107 TI - Healthy families, healthy communities. California Hospital Medical Center expands access for all things family. PMID- 12737108 TI - Lessons in grassroots lobbying. FY 2003 Omnibus Appropriations bill earmarks spending for healthcare providers. PMID- 12737109 TI - Improving performance with an executive coach. Executive coaching can enhance both individual and organizational results. PMID- 12737110 TI - Patient satisfaction with the outpatient experience. How does your organization measure up? PMID- 12737111 TI - Salvaging HAART. PMID- 12737112 TI - What's hot, what's not. PMID- 12737113 TI - Can OI prophylaxis be stopped in patients responding to HAART? PMID- 12737114 TI - Practice guidelines as disease management strategies. PMID- 12737115 TI - Metabolic effects of protease inhibitor therapy. AB - Fat redistribution (lipodystrophy) and metabolic anomalies are reported increasingly in HIV-infected patients being treated with protease inhibitors. The incidence of these side effects ranges from 5% to 75% in such patients, who often complain of spontaneous fat wasting in the face, arms, or legs, with or without central obesity. Hyperlipidemia and insulin resistance are almost always associated with lipodystrophy. We review the metabolic complications of antiretroviral therapies and discuss possible therapeutic interventions. PMID- 12737116 TI - AIDS education in Tanzania: focus on school children. AB - Tanzania is one of the countries most severely affected by the HIV/AIDS epidemic. A number of local AIDS education efforts have been launched over the past decade, several of which have yielded measurable, positive outcomes. Despite these successes, many barriers to effective dissemination of AIDS information exist, and large population groups still lack basic information on how to protect themselves from infection. Although effective school-based AIDS education programs are available, these have yet to be implemented consistently and persistently throughout the public school system, where the majority of adolescents can be reached. PMID- 12737117 TI - Are your medication records confidential? PMID- 12737118 TI - Antiretroviral therapy in the treatment-experienced patient. AB - Despite the growing number of new antiretroviral agents, planning therapy for treatment-experienced patients remains extremely challenging. Cross-resistance within the three currently available classes of drugs limits the number of sequential drug regimens that can be expected to suppress viral replication effectively. An understanding of resistance and cross-resistance, together with judicious use of resistance testing, can help clinicians design better treatment regimens for patients experiencing virologic failure while taking antiretroviral therapy. PMID- 12737119 TI - The breast-feeding dilemma and its impact on HIV-infected women and their children. AB - The rate of HIV transmission via breast-feeding ranges from 14% to 26%, depending on the timing of maternal infection. In settings where infant mortality rates from infectious diseases and malnutrition are low and relatively safe alternatives to breast-feeding are available, HIV-infected mothers should be advised not to breast-feed. Where breast-feeding by HIV-infected mothers and bottle-feeding both present serious risks of mortality, changing the conditions in which families live so that safe feeding alternatives become available must be a top priority. At the same time, these mothers need information about the relative risks and benefits of breast-feeding, early weaning, wet-nursing, and formula feeding. This article reviews the available research data and discusses critical gaps in current knowledge. PMID- 12737120 TI - Renewed hope in the treatment of hepatitis C virus infection. PMID- 12737121 TI - Treatment and prevention of tuberculosis in HIV-infected patients: CDC guidelines. PMID- 12737122 TI - Medication adherence in patients with HIV infection: a comparison of two measurement methods. AB - Two measurements of adherence, patient self-report and electronic measurement by the Medication Event Monitoring System (MEMS), were compared in a 3-month adherence study of 44 HIV-infected patients who had been placed on regimens that included protease inhibitors (PIs). The dose percentage and degree of clinically significant dosing time fluctuation were calculated monthly. The mean dose percentage by self-report versus MEMS was 97.5% versus 90.3% during month 1 of adherence monitoring, 96.5 versus 90.1% during month 2, and 98.4% versus 92.8% during month 3. Thirty-two percent of patients taking PIs and 21% of patients taking nucleoside analogues demonstrated clinically significant dosing time fluctuation. Our data confirm that self-reports of adherence overestimate true adherence behavior, and patients' self-reports of dosing times may not accurately reflect their deviation from those times. PMID- 12737123 TI - HIV treatment-associated hepatitis. AB - Combination antiretroviral therapy, as well as a variety of other medications prescribed, places patients with HIV disease at high risk for adverse drug reactions and interactions. Sever hepatitis has been reported with all of the currently available classes of antiretroviral agents. Physicians should be aware of the risk of hepatocellular injury in patients receiving antiretroviral therapy and monitor transaminase levels in those at risk. This article reviews the incidence, clinical features, risk factors, and etiology of drug-induced hepatitis with various combinations used in clinical practice. PMID- 12737126 TI - Open the door. PMID- 12737125 TI - Salvage II. PMID- 12737124 TI - Weight loss and disease progression in HIV infection. AB - Weight loss and malnutrition continue to be important issues that clinicians face when treating patients with HIV infection. In addition to specific clinical consequences, weight loss in these patients is linked to a greater risk of death and opportunistic complications. A loss of as little as 5% to 1-% of baseline body weight can be associated with a risk of death that is 2.5 times that seen in patients with HIV infection who do not lose weight. Furthermore, weight loss in patients with HIV infection can increase the risk of individual opportunistic infections by as much as 61% to 176%. Future studies may help define the prognostic implications of lipodystrophy and changes in body cell mass in patients with HIV who are taking antiretroviral therapy. PMID- 12737127 TI - Getting and keeping patients on HAART. PMID- 12737128 TI - Hiccups and vertigo in a man with AIDS. PMID- 12737129 TI - Changing incidence of mycobacterial diseases in German patients with HIV infection. AB - Before the introduction of potent antiretroviral therapy, disease caused by atypical mycobacteria was a frequent diagnosis in patients with fewer than 100 CD4+ cells/microL and could affect virtually every organ. The diagnosis was associated with a survival of usually less than 1 year, and antimycobacterial treatment could extend the life span by only several months.. However, the clinical face of the HIV epidemic has changed profoundly since 1996, most probably due ot the antiretroviral combination strategies. In this discussion, we assess the influence of the introduction of antiretroviral combination therapies on the incidence of mycobacterial infections from 1994 to 1998 in a cohort of German HIV-seropositive patients treated at an HIV outpatient clinic. PMID- 12737130 TI - Progression of autoimmune thyroiditis in an HIV-infected woman on HAART. AB - Changes in immunoregulation, among other factors, may initiate or exacerbate autoimmune thyroiditis. Strikingly high titers of antithyroid peroxidase antibodies have been found in HIV-infected patients, and, according to some studies, these increase further as HIV disease progresses. The pathogenesis of autoimmune thyroiditis is not totally understood, but activated CD4+ cells predominate in the infiltrate and are believed to be central to the process. Some investigators have postulated that endocrinologic autoimmunity might result from incomplete or unbalanced immune restoration with highly active antiretroviral therapy (HAART). The case presented here suggests progression from euthyroid Hashimoto's thyroiditis to hypothyroidism after initiation of HAART. PMID- 12737131 TI - Appetite stimulants and anabolic steroid therapy for AIDS wasting. PMID- 12737132 TI - Antiretroviral medication errors for patients with HIV infection. AB - Errors in prescribing antiretroviral agents for patients with HIV infection may lead to treatment failure, drug resistance, or drug toxicity. Several published reports describe such errors, which appear to be related to a lack of knowledge, inexperience, complexities of the antiretroviral regimens, and sound-alike and look-alike names. Clinicians caring for patients with HIV infection should be aware of the potential for prescribing errors and develop strategies to prevent them. PMID- 12737133 TI - Finding the truth: a guide to interpreting HIV clinical trials. AB - The complex social, epidemiologic, and pathologic aspects of HIV disease, the variation in the nature of the disease from person to person, and the constantly changing roster of available treatments create intense and often contradictory concerns. Clinical trials are rarely interested only in the study patients, but are designed to enable us to make inferences about a general HIV-infected population. To do this most effectively, we must confirm the internal and external validity of the study. Statistics enable us to generalize from a particular study to the broader population, but the statistical problems arising from AIDS clinical trials are unique and complex. PMID- 12737134 TI - Preventing mother-to-infant transmission of HIV in the developing world. PMID- 12737135 TI - More good news on reducing the incidence of maternal-fetal transmission of HIV and simplifying HAART. PMID- 12737137 TI - Editorial comment: Kaposi's sarcoma of the testicle: a rarity. PMID- 12737136 TI - Kaposi's sarcoma of the testicle. AB - The penis and scrotum are the genitourinary tract organs most frequently involved in Kaposi's sarcoma (KS). However, solid organ involvement is rare. A 32-year-old man with AIDS presented with cutaneous KS, which subsequently metastasized to the right testicle. PMID- 12737138 TI - The patient's perspective on life with antiretroviral treatment: results of an 887-person survey. AB - Soon after treatment with protease inhibitors became widespread, health care providers understood that adherence was essential to prevent emergence of drug resistant viral strains. However, little was known about factors influencing adherence among patients with HIV infection. To help clarify the patient's perspective, AIDES, a French AIDS service consortium, conducted a survey of 887 patients between June 1 and July 15, 1997, in its 110 chapters throughout France and through its quarterly newsletter, Remaides. Fatigue and gastrointestinal side effects were the most frequent complaints, but patients reported that their greatest difficulty was that of confronting lifelong treatment with no possibility of discontinuation. PMID- 12737139 TI - Of mice and men: cytotoxic T cells and AIDS pathogenesis. AB - CD8+ cytotoxic T lymphocytes (CTLs) represent a first line of defense against HIV infection, although their precise role in disease pathogenesis remains enigmatic. They play an important part in viral control but may also contribute to disease progression through destruction of CD4+ helper T cells. The role of CTLs in lymphocytic choriomeningitis virus (LCMV) infection in mice has been studied extensively, and the effects of CTL activity on host and virus are well defined. Although LCMV is not a retrovirus, it shares salient features with HIV, including a wide tropism, a capacity to persist, and genetic instability. The diseases caused by LCMV and HIV are linked by common immune effector mechanisms and, potentially, immunopathologies. Understanding the well-characterized immune responses in LCMV infection may therefore cast light on the role of CTLs in HIV disease. PMID- 12737140 TI - Hepatitis B and C in the context of HIV disease: implications for incarcerated populations. AB - HIV-infected inmates are often co-infected with hepatitis B and/or hepatitis C virus. To describe the burden of HIV in the incarcerated population today, one must consider the impact of co-infection. HIV may dramatically modify the course of viral hepatitis infection, especially chronic hepatitis C. The converse is uncertain: chronic hepatitis does not seem to accelerate HIV disease progression. In this article, we offer guidelines for selecting appropriate candidates for treatment among co-infected inmates. PMID- 12737142 TI - Ultraviolet (280-400 nm)-induced DNA damage in the eggs and larvae of Calanus finmarchicus G. (Copepoda) and Atlantic cod (Gadus morhua). AB - In previous work, we evaluated the effects of ultraviolet (UV = 280-400 nm) radiation on the early life stages of a planktonic Calanoid copepod (Calanus finmarchicus Gunnerus) and of Atlantic cod (Gadus morhua). Both are key species in North Atlantic food webs. To further describe the potential impacts of UV exposure on the early life stages of these two species, we measured the wavelength-specific DNA damage (cyclobutane pyrimidine dimer [CPD] formation per megabase of DNA) induced under controlled experimental exposure to UV radiation. UV-induced DNA damage in C. finmarchicus and cod eggs was highest in the UV-B exposure treatments. Under the same spectral exposures, CPD loads in C. finmarchicus eggs were higher than those in cod eggs, and for both C. finmarchicus and cod embryos, CPD loads were generally lower in eggs than in larvae. Biological weighting functions (BWF) and exposure response curves that explain most of the variability in CPD production were derived from these data. Comparison of the BWF revealed significant differences in sensitivity to UV-B: C. finmarchicus is more sensitive than cod, and larvae are more sensitive than eggs. This is consistent with the raw CPD values. Shapes of the BWF were similar to each other and to a quantitative action spectrum for damage to T7 bacteriophage DNA that is unshielded by cellular material. The strong similarities in the shapes of the weighting functions are not consistent with photoprotection by UV absorbing compounds, which would generate features in BWF corresponding to absorption bands. The BWF reported in this study were applied to assess the mortality that would result from accumulation of a given CPD load: for both C. finmarchicus and cod eggs, an increased load of 10 CPD Mb(-1) of DNA due to UV exposure would result in approximately 10% mortality. PMID- 12737141 TI - [Recessive hiccup of rare cause (compressive pleuro-pericardial lymphocele). Case report]. AB - The authors discuss the case of a 40 year-old woman, which was performed surgery for a stenosant duodenal ulcer by bilateral troncular vagotomy and hemigastrectomy, using the Pean-Billroyh 1 anastomosis. Immediately after surgery untractable hicongh gradually set in which lasted over a 14 year, till the moment when a compressive pleuropericardic lymphocele, of the right phrenic nerve was revealed, a very long time growing cyst. After the surgical removal of the lymphocele was performed, which had compressed the right phrenic nerve, singultation totally disappeared, the patient has been monitorized through periodical medical checkings for more than 4 years. The period of 14 years when the patient constantly complained of steady untractable singultation has comprised 30 admission or more, to various surgical, gastroenterology and psychiatry wards. The patient was finally considered an incurable psychotic and was medically pensioned. The authors discuss a number of possible sequels of the initial gastric surgery which could have triggered untractable singultation, without being able to control them or totally ruling out the psychotic causes. There is natural suspicion cast of either existence or beginning of this cyst simultaneous with surgery since clinical tests and explorations were able to reveal it only after 14 years. PMID- 12737143 TI - Molecular characterization and chromosomal localization of two families of satellite DNA in Prochilodus lineatus (Pisces, Prochilodontidae), a species with B chromosomes. AB - Prochilodus lineatus, an abundant species in the Mogi-Guacu river basin, represents a large part of the region's fishing potential. Karyotypic analyses based on classic cytogenetic techniques have revealed the presence of 54 meta submetacentric type chromosomes, together with the occurrence of small supernumerary chromosomes with intra and interindividual variations. This paper describes the genomic organization of two families of satellite DNA in the P. lineatus genome. The chromosomal localization these two repetitive DNA families through fluorescence in situ hybridization (FISH) demonstrated that the SATH1 satellite DNA family, composed of approximately 900 bp, was located in the pericentromeric region of a group of chromosomes of the standard complement, as well as on all the B chromosomes. The SATH2 satellite family has a monomeric unit of 441 bp and was located in the pericentromeric regions of some chromosomes of the standard complement, but was absent in the B chromosomes. Double FISH analyses showed that these two families participate jointly in the pericentromeric organization of several chromosomes of this species. The data obtained in this study support the hypothesis that the B chromosomes derive from chromosomes of the standard complement, which are carriers of the SATH1 satellite DNA. PMID- 12737144 TI - [Breast self-examination: knowledge of women attending the outpatient service of a university hospital]. AB - This is a descriptive study developed in the outpatient unit of a university maternity hospital in the city of Natal/RN. It aimed at identifying the knowledge about and alterations found during breast self-examination as well as at describing how women examine themselves. The population was formed by patients sent by other health services and the sample consisted of 109 women with age varying from 15 to 83 years. In order to collect data, the authors used a form containing identification information as well as about breast self-examination. Based on the collected data, they identified that among the 109 participants, the age of 36 (33%) of them varied from 15 to 39 years; of 58 (53%) of them from 40 to 58 years and of 15 (14%) of them from 60 to 83 years. 75% reported that they examined themselves and that they attended a health care service for correct treatment in case any alterations were detected during self-examination. PMID- 12737146 TI - Evidence-based practice in long-term care. PMID- 12737145 TI - [Food intake: who controls what?]. AB - Since the discovery of leptin and the characterization of the mechanisms leading to obesity in several animal models, considerable advance has been gained in the field of energy homeostasis. The hypothalamus plays a pivotal role in the short and long term regulatory loops that control food intake and body weight. Multiple peripheral signals, including leptin and insulin, convey information on the nutritional and metabolic status to the central nervous system. In the hypothalamus, these signals modulate several neuropeptides and intricate neuronal pathways that trigger appropriate responses of food intake and also of the autonomous nervous system and of the pituitary functions. Peripheral signals and hypothalamic neuropeptides, characterized in the last decade may represent potential targets for new pharmacological treatments of obesity. However, because of the complexity of weight regulation, such approaches may appear troublesome. PMID- 12737147 TI - Prevention of early-onset group B streptococcal disease in newborns. PMID- 12737148 TI - Immunization during pregnancy. PMID- 12737149 TI - The topical hyperbaric oxygen therapy debate. PMID- 12737150 TI - No-rinse bed bath and skin tears. PMID- 12737151 TI - Culture change and veterinary medicine. PMID- 12737152 TI - Give self-esteem a hand and improve learning. PMID- 12737153 TI - Introducing the theme--continuing veterinary medical education: where are we? PMID- 12737155 TI - Accreditation: standards for quality continuing professional development. AB - Accreditation is one way of assuring that continuing education providers are credible and competent. The Accreditation Council for Continuing Medical Education has recently modified its accreditation standards to meet the needs of the health care system and continuing medical education providers. The standards (Essential Areas and Elements) from the new accreditation system will be reviewed and discussed to illustrate one system of accreditation as a potential model for the continuing veterinary medical education professional. PMID- 12737156 TI - Mandatory continuing veterinary education requirements in the United States and Canada. PMID- 12737158 TI - One title, many hats. PMID- 12737157 TI - The Post Graduate Foundation in Australia: one model of continuing veterinary medical education. AB - The Post Graduate Foundation a has for 37 years been the major provider of continuing education opportunities for veterinarians in Australia. The purpose of this paper is to describe the Foundation's development, current offerings, and future, as well as to highlight the organization's philosophy of continuing professional development (CPD). PMID- 12737159 TI - Learning in clinical practice. PMID- 12737160 TI - Ingrown toenail relief drug products for over-the-counter human use. Final rule. AB - The Food and Drug Administration (FDA) is issuing a final rule establishing conditions under which over-the-counter (OTC) ingrown toenail relief drug products containing sodium sulfide 1 percent in a gel vehicle are generally recognized as safe and effective and not misbranded. This rule also amends the regulation that lists nonmonograph active ingredients in OTC drug products for ingrown toenail relief by removing sodium sulfide from that list. This final rule is part of FDA's ongoing review of OTC drug products. PMID- 12737161 TI - DNA, nurture and parenthood. PMID- 12737162 TI - Caveolin-I overexpression is a favourable prognostic factor for patients with extrahepatic bile duct carcinoma. AB - Extrahepatic bile duct carcinoma (EBDC) is a malignancy well known for its poor prognosis. Some clinicopathological prognostic markers have been proposed, but genetic factors have not been well investigated. We have examined expression patterns of caveolin-1, which has been shown to function as a tumour suppressor in vitro, in EBDC using immunohistochemistry. Normal tissues adjacent to the tumour cells did not show immunoreactivity for caveolin-1. A total of 22 of the 60-carcinoma tissue samples (36.7%) studied were positive for caveolin-1. Caveolin-1 immunostaining negatively correlated with the patient's age and pathological T factor (pT) in a statistically significant manner. Multivariate analysis using Cox's proportional hazards model identified caveolin-1 expression as an independent positive prognostic factor. Thus, our study suggests that caveolin-1 expression may be a useful prognostic marker for EBDC. PMID- 12737163 TI - The Human Genome Project and the courts: gene therapy and beyond. PMID- 12737164 TI - Hope fear and genetics: judicial responses to biotechnology. PMID- 12737165 TI - Mental health advance directives: having one's say? PMID- 12737166 TI - A child's right to know. PMID- 12737167 TI - Informed consent for Alzheimer's disease clinical trials: a survey of clinical investigators. PMID- 12737168 TI - The right of subjects to see the protocol. PMID- 12737169 TI - The Internet--friend or foe? A questionnaire study of orthopaedic out-patients. PMID- 12737171 TI - Making informed consent meaningful. PMID- 12737170 TI - Ethical issues in including suicidal individuals in clinical research. PMID- 12737172 TI - The value of difference. PMID- 12737173 TI - Composition and operation of selected research ethics review committees in Latin America. PMID- 12737175 TI - Nontherapeutic research, minimal risk, and the Kennedy Krieger lead abatement study. PMID- 12737174 TI - Unnecessary holes in the head. PMID- 12737176 TI - New rules for gene-transfer trials. PMID- 12737177 TI - More on the use of placebo controls. PMID- 12737178 TI - Legal rules and industry norms: the impact of laws restricting health insurers' use of genetic information. AB - Since 1991, twenty-eight states have enacted laws that prohibit insurers' use of genetic information in pricing, issuing, or structuring health insurance. This article evaluates whether these laws reduce the extent of genetic discrimination by health insurers. Using multiple data sources, it concludes that there are almost no well-documented cases of health insurers asking for or using pre symptomatic genetic test results in their underwriting decisions either before or after these laws, or in states with or without these laws. At present, health insurers are not thinking about or interested in using genetic information of this sort. Using this information is not cost effective and is not seen as contributing significantly to underwriting accuracy. However, if genetic testing information were easily available, some health insurers would consider using it in some fashion if that were legal. In the future, such information could become much more relevant to health insurers than it is now. Therefore, the major effect of these laws is to make it less likely that insurers will use genetic information in the future. Although insurers and agents are only vaguely aware of these laws, the laws have helped to convince the industry that it is not appropriate or socially legitimate to use this information. Thus, these laws have caused the insurance industry to embrace more socially oriented norms and attitudes. PMID- 12737179 TI - [History of vestibular-spatial orientation research]. AB - In order to advance our scientific endeavors in the field of vestibular-spatial orientation research, it is important to know how much research has already been carried out, and how much accurate information we currently possess. It is mandatory to have sufficient background information in order to establish a solid working hypothesis utilizing insightful approaches. Accordingly, this historical survey covers various research endeavors from the beginning of aviation until the end of 2001, including Skylab, Microgravity Vestibular Investigations, and Neurolab. PMID- 12737180 TI - Who lives? Who dies? The utility of Peter Singer. PMID- 12737181 TI - Community vs. enclave: the moral voice of community can be reflected in IRB composition. PMID- 12737182 TI - A matter for the heart: the 30th anniversary of Roe vs. Wade. PMID- 12737183 TI - [Mechanism of development of space motion sickness]. AB - Astronauts and scientists are scheduled to stay aboard the International Space Station (ISS) for three-month periods, and various problems relating to the human ability to live in space for such long periods of time, as well as medical problems, are being studied. For the crews that have operated and worked on the US space shuttle, the longest time spent in the gravity-free (accurately speaking, microgravity) environment of space has been about two weeks. Even such short periods have generated reports of various physical changes and symptoms caused by the absence of gravity. Particularly striking effects are circulatory system changes, muscular atrophy and decalcification of bones. In addition, serious problems arise during the time in space due to the symptoms of space motion sickness, caused by neuro-otological changes. Space motion sickness is experienced by nearly 70% of the first-time astronauts, in the form of such symptoms as nausea, and vomiting. It is said that space motion sickness develops at the time of adaptation of the body to the gravity-free environment. Unlike motion sickness on Earth, space motion sickness is characterized by a feeling of fullness of the head, a hot face and reduced sweating. The provocative factors include movement of the head in the pitch and roll directions and the unusual visual field. This paper reviews the research that has been carried out to date on space motion sickness, and its counter measures comprehensively, in terms of the changes in the otolith organ as well as asynchronization of information between the movements of the head and eyeballs, the deep sensations and body fluid shifts. Other problems, which may be encountered as a result of yet longer stays in space, are also discussed. PMID- 12737184 TI - Paternalism and egregious harm: Prader-Willi Syndrome and the importance of care. AB - Paternalism clashes with the usual liberal model. In this paper I argue that attempts to defend even a limited form of paternalism by liberal authors such as Joel Feinberg, Gerald Dworkin and H.L.A. Hart fail. I propose instead a bivalent model for paternalism that appeals to two separate principles: the no-harm principle and the care-principle. The notion of care discussed by contemporary feminist authors is a fundamental moral archetype that permeates history and culture. I go on to consider the case of patients with Prader-Willi Syndrome and argue that paternalism is not only permissible but imperative in cases in egregious harm. This view is enshrined in common law jurisprudence which dismisses consent as a justification in serious crime. PMID- 12737185 TI - Implications of genetic testing for the insurance industry: the UK example. AB - This report summarises the controversy of genetic tests and insurance, with a focus on the UK situation during the past decade. UK experience provides insight for future strategies to help people with genetic disadvantages make insurance provision for themselves and their families. Non-disclosure of genetic test results (already carried out for clinical purposes) may not benefit people at risk of genetic disorders or with positive genetic tests. The pressure of geneticists over a decade to prevent disclosure to insurers may have masked opportunities to use insurance to provide help for people with genetic disadvantages. To seize the opportunities now, there must be collaboration, not conflict. Politicians, geneticists, social scientists and all elements of the insurance industry can contribute to wise solutions. PMID- 12737186 TI - Difficult decisions: social and ethical implications of changing medical technology. AB - OBJECTIVES: A primary objective of this paper is to present data on subjective perceptions of health and quality of life in individuals living with early- or late-onset genetic conditions (cystic fibrosis, CF, and Huntington's disease, HD, respectively). The paper will also discuss the social and ethical dilemmas raised by advances in reproductive and testing technology, consider the different emphases in definitions in quality of life, and speculate on criteria used to make reproductive decisions following prenatal testing. METHODS: Adults suffering from CF and HD attending two regional centres in the UK took part in semi structured interviews between late 1999 and summer 2001. Self-report data on how a specific body image is constructed were also collected. Interviews were analysed using thematic qualitative analysis, and the body data were analysed using SPSS. RESULTS: Qualitative analysis uncovered themes relating to the question of how we value life, and exposed broad ethical dilemmas arising from advances in testing and treatment technology. These data are supported by findings from a body chart analysis that highlight a discrepancy between the quality of life experienced by this group of respondents and the way they believe the wider public perceives them, particularly in reference to pain. CONCLUSIONS: Although some of the data support previous findings, such as the disability paradox, other voices contradict this view. Due to their unique experiences, the accounts of individuals currently living with genetic conditions should be considered in the wider bio-ethical debates. PMID- 12737187 TI - Women and coronary heart disease: redressing the balance. PMID- 12737188 TI - Immediate follow-up after ICU discharge: establishment of a service and initial experiences. AB - Follow-up of patients discharged from the intensive care unit (ICU) is recommended as a means of service evaluation (Department of Health (2000). Comprehensive Critical Care: a Review of Adult Critical Care Services), in order to monitor the quality of the services provided Without patient follow-up, ICU staff have only 'death' or 'discharge alive from hospital' as clinical outcomes from which to judge practice performance, and limited evidence exists on which to base decisions about improvements to critical care practice (Audit Commission (1999). Critical to Success--the Place of Efficient and Effective Critical Care Services Within the Acute Hospital) To address these issues the Regional Intensive Care Unit (RICU) obtained information from patient assessment/interview on problems experienced by patients within 8-9 days (mean), following discharge from RICU A nurse-administered questionnaire was used to identify functional outcomes, nutrition and psychological issues such as anxiety and sleep disturbances Benefits of patient follow-up introduced and planned include: patient diaries for long-term patients, input from clinical psychologis, review of sedation used in RICU. PMID- 12737189 TI - Nurse-led discharge from high dependency unit. AB - High dependency care is a rapidly evolving area of critical care, with high patient turnover, which ultimately leads to high levels of pressure for beds. There is a growing trend emerging, recognizing the importance and value of nurse led initiatives in delivering effective nursing care in acute care settings. One specific nurse-led initiative this author has developed is that of nurse-led discharge (NLD) from the high dependency unit (HDU), in order to optimize the utilization of critical care beds within the HDU. An audit of the current practice was undertaken, which ultimately led to the implementation of NLD. Early experiences indicate that HDU beds are now being used more effectively. PMID- 12737190 TI - How do intensive care nurses assess patients' pain? AB - Identification and evaluation of pain in critical care patients may be difficult because of communication problems. Moreover, at present there are very few nursing studies that examine the attitudes of critical care nurses towards the assessment of patients' pain. This study was designed to determine the approach of critical care nurses towards assessing patients' pain levels, and to evaluate the problems in nursing diagnosis of those having difficulty in articulating their pain symptoms. We used a questionnaire to assess nurses attitudes to patients' pain. The study sample consisted of 91 critical care nurses who were recruited between January and February 2002. The results suggest that patient pain was considered undesirable by 44% of nurses. About 70-3% of the nurses reported resorting to administering analgesics to relieve their patients' pain. Some 57.1 % of nurses stated that they would have investigated whether the patients had really been experiencing pain, prior to administering the prescribed analgesics to patients. Some 85.7% of the sample indicated that the patients themselves would make the most accurate evaluation of their pain. The data suggested that 39.6% of nurses did not know how to evaluate pain symptoms in critical care patients suffering from complicated problems, and that 37.4% evaluated pain by monitoring the patients' behaviours. The study demonstrated that most of the critical care nurses did not know how to evaluate pain in patients having communication problems. The paper concludes by suggesting that there is a clear need to address nursing education and training with regard to evaluation and management of patients' pain whilst in critical care environment. PMID- 12737191 TI - The practice competency gap: challenges that impede the introduction of national core competencies. AB - This paper reports the findings from a study commissioned by the English National Board for Nursing, Midwifery and Health Visiting (ENB), entitled Evaluation of the Effectiveness of Educational Preparation for Critical Care Nursing. One aim of the study was to generate evidence that incorporated the national perspective on the priorities for core competencies and a national threshold standard of the ENB critical care clinical awards. Although these courses are no longer validated by the professional statutory body, the findings illuminate some key insights into the current challenges facing providers and consumers of critical care education as well as into implications for all post-registration education provision. The paper outlines how the study data were collected, how the core competendes were generated and the national consultation exercise and the levers and barriers that affect the introduction of core competencies. Other aspects of the study data are reported elsewhere (Scholes and Endacott, 2002). PMID- 12737192 TI - Ms B and legal competence: examining the role of nurses in difficult ethico-legal decision-making. AB - The case of Ms B throws up some interesting issues regarding the role of the nurse in assisting patients in making and implementing their decisions. The High Court transcript makes it clear who the voices with influence in legal matters were, and why the decisions they took were made. Absent from the myriad voices is that of the profession of nursing. Are nurses silenced by professional boundaries, the legal framework or lack of confidence? The concept of nursing advocacy is once again thrown into relief and critical questions need asking about the limits of professional nursing practice. PMID- 12737193 TI - Critical care staff rotation: outcomes of a survey and pilot study. AB - Staff rotation is defined as a reciprocal exchange of staff between two or more clinical areas for a predetermined period of time. The rationale for introducing a 'Critical Care Nurse Rotation Programme' includes important issues such as improving nurses' knowledge and skills, providing development opportunities, networking, the ability to recruit and retain nurses and the provision of a more versatile and flexible workforce. To gain the understanding of nurses' views and opinions on critical care rotation programmes, evidence was collected by means of questionnaires involving 153 critical care nurses and by undertaking semi structured interviews with four nurses. On the basis of the responses, a pilot of three Critical Care Nurse Rotation Programmes was introduced. An evaluation of the pilot project assessed participants, supervisors and senior nurses' experience of rotation and revealed very positive experiences being reported. The benefits highlighted included improving clinical skills and experience, improving interdepartmental relationships, heightened motivation and opportunities to network. The disadvantages focused on the operational and managerial issues, such as difficulties maintaining supervision and providing an adequate supernumerary period. Evidence from the survey and pilot study suggests that in the future, providing rotational programmes for critical care nurses would be a valuable strategy for recruitment, retention and developing the workforce. PMID- 12737194 TI - Optimized detection of respiratory viruses in nasopharyngeal secretions. AB - Nasopharyngeal secretions (NPS) from 121 (110 pediatric) patients with acute respiratory infections were examined for respiratory virus detection by: i) conventional virus isolation in cell cultures (CC) using HEp-2, LLC-MK2, and MDCK cells; ii) rapid virus isolation using shell vial cultures (SVC) of a mixture (MIX) of mink lung epithelial cells (Mv1Lu) and human lung carcinoma (A549) cells in comparison to LLC-MK2 and MDCK cells; iii) direct fluorescent antibody (DFA) assay on NPS cells. A pool of monoclonal antibodies (MAbs) to influenzavirus A and B, parainfluenzavirus types 1 to 3, adenoviruses and respiratory syncytial virus (RSV), as well as single MAbs to the same viruses, were used for virus identification in all three procedures. Results on 101 NPS examined in parallel showed a sensitivity of 89.5%, 73.7%, and 81.6% for CC, SVC, and DFA, respectively, with the relevant negative predictive values of 94.0%, 86.3%, and 90.0%. Specificity and positive predictive values were 100%. However, the combination of DFA and SVC gave best results in terms of sensitivity (94.7%) and negative predictive value (95.5%). Use of the new MIX cell culture system in the SVC procedure enhanced virus detection, while use of the MAb pool allowed prompt identification of negative samples and saving of reagents and time for all three procedures. The combination of DFA and SVC allows diagnosis of the large majority of viral respiratory infections within 48h, while conventional virus isolation on CC may be limited to laboratories involved in research and epidemiological studies. PMID- 12737195 TI - Epstein Barr viral load monitoring by quantitative PCR in renal transplant patients. AB - Post-transplant lymphoproliferative disorders (PTLD), ranging from lymphoid hyperplasia to clonal malignancy, are a severe complication arising in solid organ transplant patients. Their reported incidence ranges from 1 to 20%, according to factors such as type of transplanted organ and the age of recipients. A strong correlation between Epstein Barr virus (EBV) infection, the grade and type of immunosuppression and the development of PTLD has been recognized. The detection and quantification of EBV-DNA load in peripheral blood have been utilized as prognostic markers for the development of PTLD, showing a correlation between high levels of EBV-DNA in the blood and the development of PTLD. In this study, we monitored EBV viral load monthly in 15 renal transplant recipients for six months. The number of EBV-DNA copies was measured in peripheral blood mononuclear cells (PBMC) and serum samples by a quantitative PCR protocol developed in our laboratory that employes a previous screening of samples containing a significant number of viral DNA copies (> or =1000 copies/10(5) PBMC or 100 microl serum) by semi-quantitative PCR followed by a precise quantification of the only significant samples by quantitative competitive (QC)-PCR. Our 15 renal transplant patients neither developed PTLD nor had recurrent acute illnesses or acute graft rejections during the study. The results obtained in the monthly follow up of EB viral load in PBMC samples confirmed its fluctuation in asymptomatic patients reported in literature. In particular, 5/14 (35.7%) of EBV seropositive patients had an EBV-DNA load equal to 1000 EBV copies /10(5) PBMC (roughly corresponding to 10.000 copies/microg PBMC DNA), and 1/14 (7.1%) reached 5000 EBV copies /10(5) PBMC (roughly corresponding to 50.000 copies/microg PBMC DNA), at least once in our study. In the EBV seronegative patient, EBV-DNA in PBMC samples was always undetectable (less than 100 DNA copies/10(5) PBMC). EBV-DNA load in all serum samples was less than threshold value of our quantification protocol (<100 DNA copies/100 microl serum), supporting the literature data. With regard to immunosuppressive treatment, 66.7% of the six patients in whom EBV load reached values equal to or higher than 1000 DNA copies/10(5) PBMC, were on FK506 whereas only 33.3% of them were on CyA. In conclusion, further investigations are needed to better understand the role of EBV infection in the pathogenesis of PTLD in immunosuppressed patients. Given the high positive predictive value of EB viral load in peripheral blood for diagnosis of PTLD reported by several authors, and the described absence of correlation between the serological evidence of EBV reactivation and EB viral load, EBV viral load measurement in PBMC and serum samples using quantitative PCR techniques is a powerful diagnostic tool to monitor transplanted patients at risk to develop PTLD. PMID- 12737196 TI - Efficacy of an inactivated canine coronavirus vaccine in pups. AB - The efficacy of an inactivated CCoV vaccine (Duramune PC) was evaluated in four pups. Two dogs were maintained non-vaccinated. Ten days after the booster shot all the pups were challenged with a field CCoV strain administered by oro-nasal route. The vaccinated pups did not display clinical signs and shed the challenge virus for 11.25 days, evaluated by virus isolation, and 13.5 days, evaluated by PCR assay. The two non vaccinated pups displayed mild diarrhoea at day post challenge 4 and shed the challenge-virus for 14 and 15 days respectively, by virus isolation, and for 22 and 24 days respectively, by PCR assay. PMID- 12737197 TI - A molecular method for the recovery and identification of enteric virus in shellfish. AB - In this paper we report the results of an investigation into the presence of enteric viruses in shellfish from the waters around Sardinia. Twenty two samples of shellfish were examined using a rapid and sensitive technique to concentrate and detect viral RNA in shellfish tissues. After recovery of viral particles, RNA was extracted, transcribed into cDNA and amplified using "nested PCR". Testing with enterovirus-specific RT-PCR produced positive results in over 13% of specimens. The virus detection procedure appears to be effective. In some circumstances it could be a better test of water quality than conventional monitoring techniques. PMID- 12737198 TI - Various morphological aspects of Escherichia coli lysis by RNA bacteriophage MS2 observed by transmission and scanning electron microscopes. AB - Escherichia coli cell lysis profiles induced by the cloned lysis gene from RNA bacteriophages MS2 were presented. Transmission electron micrographs showed that ballooning structures appeared on the cell surfaces and the others were leaking materials through the cell wall. Harsh rupture of the host cell was also observed. Scanning electron micrographs revealed many extruded structures from the cells. For the scanning electron microscopy, plastic sheets coated with calf dermis collagen and SEMPORE filters were successfully used to collect the samples and for various chemical treatments. The lysing cells by bacteriophage MS2 lysis gene observed in transmission and scanning electron micrographs showed various morphological aspects of the intermediates in the lysing process. PMID- 12737199 TI - Staphylococcus aureus subcutaneous abscess complicating acupuncture: need for implementation of proper infection control guidelines. AB - We report a case of Staphylococcus aureus subcutaneous abscess centered over the Jizhong acupuncture point (DU 6) which lies along the Du (Back midline) meridian after acupuncture at the corresponding acupuncture point for low back pain. The patient recovered after surgical debridement and drainage and 5 weeks of cloxacillin therapy. Among the 16 anecdotal case reports of pyogenic bacterial infections complicating acupuncture described in the English literature (MEDLINE Search 1996-2002), S. aureus was documented to be the causative agent in 9 (56%). Three patients had septic arthritis, 2 had chronic osteomyelitis, 2 had abscess formation, 1 had chondritis, and 1 had infective endocarditis. Five patients had S. aureus bacteremia. All patients who recovered required prolonged antibiotic treatment of 5-6 weeks, and 6 required drainage and/or debridement. Overall, 3 patients (30%) died. S. aureus causes significant morbidity and mortality in patients who receive acupuncture treatment. More resources should be spent on implementation of proper infection control guidelines, as the money lost due to prolonged hospitalization and medication would far exceed that used for implementation. PMID- 12737200 TI - A methicillin-resistant Staphylococcus aureus outbreak in a new University hospital due to a strain transferred with an infected patient from another city six months previously. AB - Kocaeli University Medical School was established in 1995. The first methicillin resistant Staphylococcus aureus isolate was detected two years later in a patient transferred from a different city. Six months after this, we detected a small MRSA outbreak in the intensive care unit involving four patients, two of whom had bacteremia, and a staff nasal carrier. All isolates, including the first, appeared to be a single outbreak strain, demonstrated by pulsed field gel electrophoresis profiles which different by at most two bands, identical randomly amplified polymorphic DNA profiles, and identical coagulase gene types by PCR. Antibiogram were identical except that one isolate was additionally resistant to cotrimoxazole. These results show that MRSA isolates can spread between hospitals with infected or colonized patients and can apparently persist in the hospital for six months without causing infection. Screening of asymptomatic patients on wards affected by MRSA or transferred from other hospitals may be helpful in controlling these infections. PMID- 12737201 TI - Evaluation of susceptibility of Mycobacterium bovis to antituberculous drugs by radiometric BACTEC 460TB system. AB - Susceptibility of Mycobacterium bovis strains to antituberculous drugs (isoniazid and rifampin) was detected by radiometric BACTEC 460TB system. M.bovis strains were isolated from tissue samples showing tuberculous lesions collected at an abbattoir from cattle belonging to 47 tuberculosis outbreaks occurring in Northern Italy in 1995-1999. Forty-six out of 61 strains (75.4%) resulted susceptible to both isoniazid and rifampin. Thirteen strains (21.3%) were resistant to isoniazid only. No strains showed resistance to rifampin only. Two strains (3.3%) resulted resistant to both drugs, showing antituberculous multidrug-resistance. Given the compulsory eradication program of bovine tuberculosis by elimination of infected animals and the ban on antituberculous drug treatments in animals, detection of resistant M. bovis strains appears of great interest. PMID- 12737203 TI - Formalin fixing and paraffin embedding may lead to extra band development in PCR. AB - The molecular biological analysis of infectious agents requires the availability of a reliable source of microorganisms to be used to recover DNA. Clinical samples can be obtained directly from infected patients or can be propagated using in vitro or in vivo systems. However, repeated sampling from patients is not always possible as the procedure may be invasive or unpleasant, or it is not possible to catch the same agent at the time of second sampling. Moreover, the techniques used may also produce false-positive and false-negative results. We therefore studied the impact of formalin-fixing and paraffin embedding on tissue sampling, and the methodologies such as DNA isolation and PCR amplification of DNAs from archival materials in the diagnosis of Mycobacterium tuberculosis. PCR analyses were done according to standard methods with some modifications. Demonstration of mycobacteria was successful both in tissue sections of the formalin-fixed lymph nodes and in stained fresh materials from patients. However, the results showed the presence of two extra bands in the gel. We accounted for extra band development due to the harshness of the methodology used to isolate nucleic acids from formalin-fixed and paraffin embedded tissue samples or the nature of the fixation procedure, or because of the time passed during storage in which alteration in the chromosomal DNA would take place. Thus, if disease- and tissue specific morphological features, such as sample size, type of fixation, and intralesional heterogeneity are ignored, errors because of sampling and methodologies used may lead to false-positive and false-negative results. PMID- 12737202 TI - Streptococcus pneumoniae nasopharyngeal colonization in young healthy children: rate of carriage, serotype distribution, and antibiotic resistance. AB - The nasopharyngeal colonization rate of Streptococcus pneumoniae and its antibiotic susceptibility was determined in a given population of 317 young children (ages 1-7 years) in the area of Bari, Italy. 18.29% of the cultures were positive for S. pneumoniae. 8.62% of the strains were intermediately resistant to penicillin. Erythromycin-(65.51%) and cotrimoxazole-(17.24%) resistance was also observed whereas all the strains resulted uniformely susceptible to cefotaxime and ceftriaxone. The high rate of nasopharyngeal carriage of Streptococcus pneumoniae along with the resistance to antibiotics widely used in the community suggests the importance of epidemiological surveillance as well as the application of new vaccine strategies. PMID- 12737204 TI - Sanitary quality of marine sediments and sands from an Italian beach. AB - Microbiological surveys were carried out on marine sands and sediments collected at a sandy beach along a coastal area close to Rome, Italy. Low densities of faecal indicator bacteria were recovered, and among them enterococci outnumbered the coliforms. The group of staphylococci was in a fairly constant concentration throughout the period of sampling. Some statistically significant correlations were calculated between yeasts and moulds, Escherichia coli and enterococci and between the latter and Clostridium spores. The data obtained could be a reference point for further studies. PMID- 12737205 TI - Immunomagnetic assay, classic culture method and fermentation tube test in the recovery of Escherichia coli O157 from sewage. AB - The diffusion of E. coli O157 was studied in the effluents of a large sewage treatment plant. Three methods were used: the Culture Method, Immunomagnetic Separation (IMS) and Fermentation Tube Test. Vero cells assay and PCR were used to assess toxin production and the presence of virulence genes. E. coli O157 were only detected using IMS and the Fermentation Tubes Test: one strain in raw sewage and 2 in a sample of disinfected effluent. The latter resulted positive for the production of verocytotoxin and possessed genes coding for vt2 and type gamma intimin. The best results were obtained with SMAC-CT-BCIG medium. The use of various different recovery techniques confirmed the difficulty involved in isolating E. coli O157 from water samples. PMID- 12737206 TI - Incidence and characterisation of Aeromonas spp. in environmental and human samples in southern Italy. AB - The incidence and characterisation of Aeromonas species in human and environmental samples in southern Italy were investigated. The results emphasize that 12.3% of the 210 examined patients carried Aeromonas spp. in their faeces. These results underline the need to include Aeromonas spp. in the list of routinely analysed enteropathogens in all diarrhoeal stool samples, especially in children below 10 years of age, elderly persons and immunocompromised patients. PMID- 12737207 TI - Treatment of PBMC with killed Helicobacter pylori subverts the environment of inflammatory cytokines. AB - It is well known that inflammation induced by Helicobacter pylori is characterized by the local production of cytokines and chemokines. In the present study, we analyse the kinetics of MCP-1, IL-12 and IL-4 induction during the interaction of peripheral blood mononuclear cells with killed and/or live H. pylori. Our results demonstrate that live H. pylori does not induce IL-4 release whereas it stimulates MCP-1 and IL-12 production. In addition, the neutralization of IL-12 with monoclonal antibodies determines a lower MCP-1 release. These data demonstrate that MCP-1 production is in part supported by IL-12 induced by live H. pylori. On the contrary, killed H. pylori stimulates the IL-4 but not MCP-1 and IL-12 production. The combined treatment with killed and live H. pylori upregulates the IL-4 production and at the same time downregulates IL-12 and MCP 1 production. PMID- 12737208 TI - Three-dimensional analysis of endosseous palatal implants and bones after vertical, horizontal, and diagonal force application. AB - The effects of bite and orthodontic forces exerted on endosseous palatal implants are not completely understood. This applies especially to the biomechanical properties inherent in the different implant geometries and resulting bone remodelling reactions on the one hand, and to the influence on the direction and magnitude of the applied forces on the other. The results of this study should help in the selection of implants for clinical use. Three types of endosseous implants (all 9 mm in length and 3.3 mm in diameter, made of titanium) were used for this investigation. Type 1 was a simple, cylinder-shaped implant; type 2 a cylinder-shaped implant with a superperiosteal step; and type 3 a cylinder-shaped implant, subperiosteally threaded, with a superperiosteal step. The load on the implant was investigated under three conditions of bite and orthodontic forces from 0.01 to 100 N (vertically, horizontally, and diagonally). The study results were calculated by means of a finite element (FE) method. Vertical loading caused bone deformation of more than 600 microeps at the simple implant. The largest deformations at this load were found in the trabecular bone with all three implant geometries. However, trabecular bone deformation was reduced by a superperiosteal step. Horizontal loading of the implants shifted the deformation from the trabecular to the cortical bone. Furthermore, a large deformation was measured at the transition from cortical to trabecular bone. The smallest deformations (less than 300 microeps) were found for implants with a superperiosteal step and diagonal loading (type 2). The use of threads provided no improvement in loading capacity. All implant types investigated showed good biomechanical properties. However, endosseous implants with a superperiosteal step had the best biomechanical properties under low loads. Thus, the trend should be to optimize the design of implants by producing small implants with additional anchorage on the bone surface. PMID- 12737209 TI - Photo-elastic stress analysis of initial alignment archwires. AB - Photo-elastic models replicating a lower arch with a moderate degree of lower incisor crowding and a palatally displaced maxillary canine were used to evaluate the stresses transmitted to the roots of the teeth by initial alignment archwires. Six initial alignment archwires were compared, two multi-strand stainless steel wires, two non-super-elastic (stabilized martensitic form) nickel titanium wires, and two stress-induced super-elastic (austenitic active) nickel titanium wires. Three specimens of each archwire type were tested. Analysis of the photo-elastic fringe patterns, in the medium supporting the teeth, revealed that the non-super-elastic nickel titanium archwires produced the highest shear stresses (P = 0.001). However, the shear stresses generated by the super-elastic alignment archwires and the multi-strand stainless steel archwires were very similar (P = 1.00). These results show that even in situations where large deflections of initial alignment archwires are required, super-elastic archwires do not appear to have any marked advantage over multi-strand stainless steel alignment archwires in terms of the stresses transferred to the roots of the teeth. PMID- 12737210 TI - Gonial angle changes after bilateral unidirectional mandibular distraction in sheep. AB - It was the aim of this study to investigate radiologically the distraction site in the gonial angle area after three different protocols of unidirectional mandibular distraction in sheep. Bilateral mandibular distraction was performed in nine sheep, and one other sheep served as a sham-operated control. In the nine experimental sheep, three different distraction protocols were used. In three sheep, an oblique corticotomy line was applied and the distractors were positioned perpendicular to the corticotomy line. In three other sheep, a horizontal corticotomy line was applied and the distractors were also positioned perpendicular to this corticotomy line. In the remaining three sheep, again, a horizontal corticotomy was performed, but with an angular positioning of the distractors relative to this corticotomy line. Distraction of 20 mm was performed over 20 days at a rate of 1 mm per day. After 3 weeks of retention the sheep were sacrificed, the mandibles were dissected and hemisectioned, and standardized hemimandibular contact radiographs were made of the specimens. The distracted gaps were traced from the radiograph and the gonial angle was measured. The displacements of the distracted parts were measured, the gonial angles were determined, and the values were compared with four additional control non distracted sheep mandibles. The results show that there was a very large variability in the distracted gaps as well as in the gonial angle changes after distraction, and it was not possible to recognize any of the three different distraction protocols radiographically after distraction. Although performed under standardized conditions it is difficult to reproduce intended mandibular corticotomy lines in sheep. PMID- 12737211 TI - Stresses on the cervical column associated with vertical occlusal alteration. AB - The biomechanical effects on cervical vertebral columns (C1-C7) during mastication were calculated using a three-dimensional (3D) finite element method. To verify the biomechanical influences of vertical occlusal alteration to the cervical column, three finite element models (FEM) showing a normal (model A), a steep (model B), and a flat occlusal plane (model C) were constructed. The occlusal stress distribution showed various patterns for the three models; the stress extended to the anterior area as the occlusal plane became steeper. The plots of the stresses on the mid sagittal section of the cervical columns showed different patterns for the three models; the stress converged at the odontoid process in models A and B, whereas the stresses at C7 in model B tended to decrease compared with model A. Concentrated stress was observed at C5 in model C, supporting the hypothesis that vertical occlusal alteration could influence stress distribution in the cervical columns. PMID- 12737213 TI - Quantifying enamel demineralization from teeth with orthodontic brackets--a comparison of two methods. Part 1: repeatability and agreement. AB - The aim of this investigation was to compare the repeatability of measuring enamel demineralization surrounding an orthodontic bracket using two techniques: computerized image analysis from digitally converted photographic slides and quantitative light-induced fluorescence (QLF). Fifteen human molars were halved and shaped to look like incisors. The teeth were individually numbered and orthodontic brackets bonded to the buccal surface. The crowns were covered with acid resistant varnish, except for windows approximately 1.5 x 3 mm adjacent to the gingival, occlusal, mesial, and distal edges of the bracket. The windows were variously exposed to a demineralizing gel for 0, 3, 7, or 14 days, and the acid resistant varnish was removed. Standardized photographic slides and QLF images of the teeth were taken. These were repeated after 1 week. The slides were converted to grey scale digital format and analysed using Image-Pro Plus 3.0. The QLF images were stored, processed, and analysed using customized software. All images were recoded for blind analysis. The four surfaces of the bracket were inspected and only areas of suspected demineralization were analysed. This was repeated after 1 week. The limits of agreement and mean difference between repeat readings of the area of demineralization were similar for both techniques (-0.04 +/- 0.43 for photographs and -0.10 +/- 0.63 for QLF). Mean grey level (photographs) and mean loss of fluorescence from that area (deltaF) (QLF) showed acceptable limits of agreement. The Intra Class Correlation (ICC) was below 0.81 for the measurement of area from QLF, suggesting that random error needs to be reduced. There was evidence of systematic bias for the repeat readings of the grey levels from the photographs (P < 0.001). Enamel demineralization surrounding an orthodontic bracket can be measured reproducibly using these two techniques. PMID- 12737214 TI - Quantifying enamel demineralization from teeth with orthodontic brackets--a comparison of two methods. Part 2: validity. AB - This is Part 2 of an in vitro study investigating two techniques for recording and quantifying demineralization surrounding orthodontic brackets. In Part 1 the repeatability of computerized image analysis from digitally converted photographic slides and quantitative light-induced fluorescence (QLF) was explored. In Part 2 of the investigation the validity of each technique was examined. Thirty halved human molars, shaped to look like incisors and with an orthodontic bracket bonded to the buccal surface were used. A small area of each of the four edges of the bracket was variously exposed to a demineralizing gel for 0, 3, 7, or 14 days. Images of the teeth were taken and analysed using the two techniques. Repeat images and analysis were carried out blind. The sensitivity, specificity, positive, and negative predictive values were calculated. A negative result confirmed that there was no demineralization present in the majority of cases. A positive result was less reliable particularly for the occlusal and gingival regions. The 7- and 14-day demineralization patterns were detected every time using the photographic technique. The discrimination of the 0- and 3-day patterns was less reliable. The results of this study were extrapolated to allow for the prevalence of the condition following orthodontic treatment, and the implications of this for a putative clinical trial are discussed, together with the advantages and disadvantages of each method. Either technique would be applicable to use in a clinical trial. PMID- 12737212 TI - Molecular mechanisms in calvarial bone and suture development, and their relation to craniosynostosis. AB - The development and growth of the skull is a co-ordinated process involving many different tissues that interact with each other to form a complex end result. When normal development is disrupted, debilitating pathological conditions, such as craniosynostosis (premature calvarial suture fusion) and cleidocranial dysplasia (delayed suture closure), can result. It is known that mutations in the fibroblast growth factor receptors 1, 2, and 3(FGFR1, 2, and 3), as well as the transcription factors MSX2 and TWIST cause craniosynostosis, and that mutations in the transcription factor RUNX2 (CBFA1) cause cleidocranial dysplasia. However, relatively little is known about the development of the calvaria: where and when these genes are active during normal calvarial development, how these genes may interact in the developing calvaria, and the disturbances that may occur to cause these disorders. In this work an attempt has been made to address some of these questions from a basic biological perspective. The expression patterns of the above-mentioned genes in the developing mouse skull are detailed. The microdissection and in vitro culture techniques have begun the task of identifying Fgfrs, Msx2, and Twist interacting in intricate signalling pathways that if disrupted could lead to craniosynostosis. PMID- 12737215 TI - Restoration of mechanical strength and morphological features of the periodontal ligament following orthodontic retention in the rat mandibular first molar. AB - Biomechanical properties and morphological features of the periodontal ligament (PDL) in the rat mandibular molars were examined during orthodontic retention. Seventy-three male rats of the Wistar strain, 8 weeks of age, were used for biomechanical analysis and six rats for morphological analysis. An elastic band was inserted between the mandibular first and second molars for 4 days; after removal of the elastic band the interdental space was filled with resin for 4 and 8 days. The maximum shear stress, tangent modulus, and failure strain energy density of the PDL of the first molar in the experimental animals decreased markedly following application of an orthodontic force. They increased rapidly and were restored completely to the control levels by the 8th day after retention. Light microscopy showed severe compression and extension of the PDL in the experimental animals on the 8th day after retention. Birefringent collagen fibre bundles running across the compressed and expanded PDL were observed, although they appeared to be thinner with less insertions into the alveolar bone or cementum in the experimental animals than in the controls. This suggests that the periodontal collagen fibres were partially reorganized and rearranged during retention. The reorganization and rearrangement of periodontal collagen fibres seemed to be partly related to the restoration of mechanical strength of the rat molar PDL during the 8 days of retention. PMID- 12737216 TI - Social inequality and discontinuation of orthodontic treatment: is there a link? AB - The aims of this study were to investigate the effects of social inequality on the likelihood of patients discontinuing orthodontic treatment, and to determine which, if any, indicators of social inequality are of greater relevance. In this retrospective study of English and Welsh General Dental Services (GDS) cases, consecutive 'discontinued' cases collected at the Dental Practice Board (DPB) during 1990-91, were compared for age, treatment modality, and measures of social inequality, with a 2 per cent sample of cases contemporaneously submitted as 'complete'. Three deprivation indices, and occupation-based social class spectra of neighbourhoods, were compared between the groups. A model was sought to predict discontinuation/completion using logistic regression analysis. The discontinued sample represented lower social stratum spectra for home and practice areas under all indicators tested, and the subjects were a little older at the start of treatment. Fewer were treated by orthodontically qualified practitioners or with fixed appliances, but more with extra-oral traction. Occupation-based classification (patient's home) and the Carstairs Index (practice area) were selected by the analysis as explaining more of the variation than other measures of social inequality, but the model failed to predict the discontinued cases. Lower social class may be a risk factor for discontinuation of orthodontic treament, but is not a predictor for it. Patients should be considered for, and counselled about, orthodontic treatment on an individual basis. Occupation-based social classifications and the Carstairs Index may be a little more sensitive to orthodontic applications than other indicators of social inequality. PMID- 12737218 TI - The effect of prostaglandin E2 and calcium gluconate on orthodontic tooth movement and root resorption in rats. AB - Possible modifications in orthodontic tooth movement (OTM) and root resorption as a result of local injections of prostaglandin E2 (PGE2) alone and with calcium gluconate (Ca) formed the aim of the present study. Twenty-four 8-week-old male Wistar rats were selected and randomly divided into three groups of eight. Both quadrants of the upper jaws of the first group of animals were used; therefore this group comprised two groups: control and normal. The upper left first molars of these eight animals were not placed under orthodontic force and received no injection, to serve as the normal group, considered for root resorption comparison only. The control group had localized submucosal injections of normal saline on the buccal side of the upper right first molar. In the third group, 0.1 ml of 1 mg/ml PGE2 was injected at the same site and the fourth group received an intraperitoneal injection of 200 mg/kg Ca (10%) in addition to the PGE2. All the injections were performed on days 0 and 7. The orthodontic appliance consisted of a closed coil spring ligated to the upper right first molar and incisor, exerting a force of 60 g during the 21-day experimental period, after which the animals were sacrificed. Palatal halves were removed for histological examination and for calculation of the amount of root resorption. Statistical analysis of data showed a significant (P < 0.05) acceleration in OTM after PGE2 injection compared with the control group. The addition of Ca reduced OTM but a significant increase (P< 0.05) was still recorded. A significant difference (P < 0.05) in root resorption was only observed between the PGE2 and normal groups. The findings show the importance of calcium ions working in association with PGE2 in stabilizing root resorption while significantly increasing OTM. PMID- 12737217 TI - Longitudinal cephalometric standards for the neurocranium in Norwegians from 6 to 21 years of age. AB - The purpose of this study was to establish and describe normative cephalometric standards of the neurocranium (theca cranii and cranial base) for Norwegian males and females from 6 to 21 years of age using lateral cephalograms. The subjects included 35 males and 37 females from the Oslo University Growth Archive with lateral cephalograms taken every third year from 6 to 21 years of age. The total number of lateral cephalograms was 194 from males and 200 from females. All subjects were Caucasian, all had normal occlusion and no apparent facial disharmony, and none had undergone orthodontic therapy. Nineteen measurements and three indices of the neurocranium were analysed longitudinally. Comparisons between the various parameters in the neurocranium of males and females in each age group were performed using the Student's t-test. The size of the neurocranium of females was smaller than that of males throughout the observation period and the differences increased with age, particularly the diameter of the neurocranium (n-l), length of the neurocranium (n-opc), anterior cranial base length (n-s), and posterior cranial base length (s-ba). The cephalometric standards of the neurocranium established in this study can be used as a reference material in investigations of individuals with various craniofacial aberrations and syndromes. PMID- 12737219 TI - Prediction of outcomes of secondary alveolar bone grafting in children born with unilateral cleft lip and palate. AB - The aim of this cross-sectional clinical outcome study using retrospective data capture of treatment histories was to examine the characteristics of children born with unilateral cleft lip and palate (UCLP) in the United Kingdom (UK) who were not grafted at the appropriate age or who had an unsuccessful secondary alveolar bone graft. The subjects were born with complete non-syndromic UCLP between 1.4.82 and 31.3.84 and were aged between 12.0 years and 14.7 years at the time of data collection under the care of 48 cleft teams. The success of secondary alveolar bone grafting was assessed using a modification of the Bergland index. There were no independent predictors for unmet bone grafting need. The outcome of secondary alveolar bone grafting was assessed for 164 subjects; 90 (55%) had a successful first graft. Non-Caucasian (P = 0.037) and increasing age at grafting (P = 0.007) were risk factors for poor outcome. After adjustment for other risk factors, increased age at grafting was independently associated with having a seriously deficient or failed graft (OR = 1.03; 95% CI 1.01-1.06 P = 0.036). All the non-Caucasians in this sample had an unsatisfactory graft. Increasing age in months at grafting and ethnicity are predictors for poor outcome of secondary alveolar bone grafting in children born with UCLP in the UK. PMID- 12737220 TI - The immunologists' debt to the chicken. AB - The immune system of the chicken is an invaluable model for studying basic immunology and has made seminal contributions to fundamental immunological principles. Graft versus host responses and the key role of lymphocytes in adaptive immunity were first described in work with chicken embryos and chickens. 2. Most notably, the bursa of Fabricius provided the first substantive evidence that there are two major lineages of lymphocytes. Bursa-derived lymphocytes, or B cells, make antibodies while thymus-derived, or T cells, are involved in cell mediated immune responses. 3. Gene conversion, the mechanism used by the chicken to produce its antibody repertoire, was first described in the chicken and requires the unique environment of the bursa. Subsequently it has been shown that some mammals also use gene conversion. 4. The chicken's Major Histocompatibility Complex (MHC), the first non-mammalian MHC to be sequenced, is minimal, compact and some 20-fold smaller than that of mammals. Uniquely, the chicken MHC is strongly associated with resistance to infectious diseases. 5. The first attenuated vaccine was developed by Louis Pasteur against a chicken pathogen, fowl cholera, and the first vaccine against a natural occurring cancer agent, Marek's disease virus, was developed for the chicken. 6. Vaccination of chick embryos on the 18th d of incubation, another breakthrough using chickens, provides protection early after hatching. In ovo vaccination now is widely practised by the poultry industry. 7. Evidence that widespread and intensive vaccination can lead to increased virulence with some pathogens, such as Marek's disease virus and infectious bursal disease virus, was first described with chicken populations. It warns of the need to develop mo resustainable vaccination strategies in future and provides useful lessons for other species, including in the human population. 8. Recombinant DNA technologies now provide the opportunity for the rational design of new vaccines. Such vaccines could contain the protective immunogenic elements from several pathogens and immunomodulatory molecules to direct and enhance immune responses so providing improved protection. The important thing will be to design vaccines that are sustainable and do not drive pathogens to ever-increasing virulence. PMID- 12737221 TI - Behaviour and welfare of individual laying hens in a non-cage system. AB - 1. A leg band containing a transponder was fitted to 80 birds in a perchery containing 1,000 birds. 2. The transponder emitted a unique identification number when a bird walked on one of 8 flat antennae on the floor. The recording apparatus was used to measure the amount of time that each of the tagged birds spent on the slatted and littered areas in a 6-week period. 3. Some birds spent long periods of time on the slats, possibly as a means of avoiding repeated attacks. Duration on the slats was greatest in birds with the worst (as opposed to better) feather scores of the head, back and tail regions. 4. Birds that spent long periods on the slats were lighter than other birds at both 39 weeks of age and 72 weeks of age and had greater back, head and tail feather damage, consistent with these birds being victims of pecking. 5. Tagged birds received a social avoidance test outside the perchery at 39 weeks of age, which suggested that birds retreated to the slats in response to pecks rather than just to close proximity to other birds. 6. The failure to find that duration on the slats was related to anatomical indicators of stress (liver, spleen and bursa of Fabricius) suggests that retreating to the slats following pecking attenuates physiological stress responses. 7. We conclude that the provision of areas where birds in a large group can avoid pecking may improve the welfare of a minority of victimised birds. PMID- 12737222 TI - Purification of avian circulating primordial germ cells by nycodenz density gradient centrifugation. AB - 1. Primordial germ cells (PGCs) are the progenitor cells for gametes. In aves, PGCs show a unique migration pathway, that is, they circulate temporarily through the bloodstream during early development. 2. In this study we developed a method to purify circulating primordial germ cells (cPGCs) in quail and chicks by Nycodenz density gradient centrifugation. 3. The process consisted of primary and secondary purification. In primary purification, cPGCs were enriched at the interface of 8 and 12% Nycodenz fractions. In secondary purification, cPGCs were harvested from 8% Nycodenz fraction at a purity of 90% and from 10% Nycodenz fraction at a purity of 70%. The recovery rate of cPGCs was over 70%. 4. This method would facilitate research on cPGCs' culture and the production of transgenic birds using cPGCs. PMID- 12737223 TI - Introduction of exogenous DNA into gonads of chick embryos by lipofection and electroporation of stage X blastoderms in vivo. AB - 1. In order to introduce exogenous DNA into gonads of chick embryos, stage X blastoderms of freshly laid and unincubated eggs were transfected by lipofection and electroporation in vivo. 2. The introduced DNA, green fluorescence protein (GFP) gene, was efficiently expressed in the blastoderms incubated for 24 h (78.8%, 78/99). 3. The GFP gene was present in most of the embryonic bodies and extra-embryonic membranes died by d 10 of incubation, when analysed by polymerase chain reaction. On d 16 to 20 of incubation, the GFP gene was detected in 7.0 to 20.9% of embryos in the heart, liver, stomach and brain, but not in the sartorius muscle. For the gonads, the GFP gene was detected in 22.2% (6/27) of the testes and 6.3% (1/18) of the ovaries examined. 4. These results suggest that it is possible to introduce exogenous DNA into gonads of chick embryos by lipofection and electroporation of stage X blastoderms in vivo. PMID- 12737224 TI - Inheritance of reproductive traits of female common ducks (Anas platyrhynchos) in pure breeding and in inter-generic crossbreeding with muscovy ducks (Cairina moschata). AB - 1. Genetic parameters of reproductive traits were estimated in a population of common duck, in purebreeding and crossbreeding (with Muscovies) insemination systems. A total of 989 females were studied over three generations as well as 4025 purebred offspring and 4,125 male mule offspring. 2. Traits studied were age at first egg, total number of eggs laid until the age of 48 weeks, fertility and hatchability rates in pure and crossbreds, weight at 6 and 30 weeks of age, average egg weight and body weight of the male mule ducks at 6 weeks of age. 3. Heritability estimates were found to be medium range for reproductive traits (0.15 to 0.47). Heritability value for fertility or hatchability in crossbreds was twice as high as in purebreds (0.32 vs 0.15 for fertility; 0.36 vs 0.16 for hatchability). 4. Fertility in purebreeding and in crossbreeding were two different traits (r(g) = 0.49) while hatchability displayed a high genetic correlation between breeding systems (r(g) = 0.88). 5. Genetic correlations with number of hatched mule ducks were medium or high and favourable. Genetic correlations between reproductive traits and weights were low (< 0.36), the most related trait being the body weight of the male mule duck at 6 weeks of age. PMID- 12737225 TI - The effect of chronic feeding of diacetoxyscirpenol and T-2 toxin on performance, health, small intestinal physiology and antibody production in turkey poults. AB - 1. The effects of feeding T-2 toxin or diacetoxyscirpenol (DAS) at levels up to 1 ppm for 32 d on performance, health, small intestinal physiology and immune response to enteral and parenteral immunisation were examined in young poults. 2. Slight improvement in growth was observed in some groups of poults fed T-2 or DAS mycotoxins for 32 d, with no change in feed efficiency. Feeding both T-2 and DAS resulted in oral lesions which had maximal severity after 7-15 d. 3. Mild intestinal changes were observed at 32 d but no pathological or histopathological lesions were found. Both mycotoxins altered small intestinal morphology, especially in the jejunum where villi were shorter and thinner. In addition, both DAS and T-2 mycotoxins enhanced the proportion of proliferating cells both in the crypts and along the villi. Migration rates were reduced in the jejunum of poults fed T-2 toxin but did not change in the duodenum or in poults fed DAS. 4. No significant effects of T-2 or DAS were observed on antibody production to antigens administered by enteral or parenteral routes. 5. This study indicates that tricothecene toxins at concentrations of up to 1 ppm for more than 30 d influenced small intestinal morphology but did not affect growth or antibody production. PMID- 12737226 TI - Dispersal of micro-organisms in commercial defeathering systems. AB - 1. The extent of cross contamination between carcases and the dispersal of micro organisms to the environs during defeathering was measured in a commercial processing plant. 2. Defeathering reduced the numbers of a marker organism, a nalidixic acid-resistant strain of Escherichia coli K12, on inoculated carcases but dispersed the organism on to preceding and following carcases. 3. The pattern of microbial dispersal during defeathering was similar for naturally occurring bacteria on the carcase, for example, total aerobic counts and counts of presumptive coliforms, suggesting that the marker organism mimics the natural situation realistically. 4. The majority of feathers, together with micro organisms, were removed during the first 10 s of the defeathering process, which was completed in 45 s, indicating that control measures to minimise cross contamination would be most effective if applied in the early stages of the process. 5. The method of defeathering used by the machine influenced the pattern of microbial dispersal and the extent of cross contamination to other carcases on the same processing line. PMID- 12737227 TI - Effect of enzyme preparation containing xylanase and beta-glucanase on performance of laying hens fed wheat/barley- or maize/soybean meal-based diets. AB - 1. A commercial enzyme preparation (Quatrazyme HP) containing xylanase and beta glucanase was examined in two laying hen experiments with wheat/barley- or maize based diets. The activities of other enzymes were measured also. Starch, cell wall contents and effects of Quatrazyme HP on in vitro viscosity of wheat, barley, maize and soybean meal were determined. 2. In the first experiment, 90 ISA Brown laying hens at 28 weeks of age were given a wheat/barley basal diet with or without 20mg of Quatrazyme HP, which provided 560 and 2,800 IU of xylanase and beta-glucanase/kg diet. In the second experiment, 66 ISA Brown laying hens at 45 weeks of age were given a maize/soybean meal basal diet with or without 20 mg of Quatrazyme HP/kg diet for 9 weeks. Egg production, egg weight, egg mass, feed conversion ratio and change in body weight were recorded as response criteria. 3. There was a significant improvement in feed conversion ratio with enzyme supplementation. Birds given an enzyme-supplemented diet gained 86 g while those fed on the unsupplemented diet lost 103 g of their body weight by the end of the experiment. 4. The enzyme preparation did not affect either egg production, egg weight or egg mass of birds fed on the maize/soybean meal diet. However, a significant improvement in feed conversion ratio was detected. Birds on either the supplemented or unsupplemented diet exhibited an increase in their body weight at the end of the experiment. 5. Addition of xylanase and beta glucanase decreased in vitro viscosity of wheat, barley, maize and soybean meal. This effect was greater for wheat and barley than for maize and soybean meal. 6. It was concluded that the beneficial effect of using an enzyme preparation containing xylanase and beta-glucanase is not limited to wheat/barley-based diets but also occurs with maize/soybean meal-based diets. PMID- 12737228 TI - Effect of whole and demucilaged linseed in broiler chicken diets on digesta viscosity, nutrient utilisation and intestinal microflora. AB - 1. A study was undertaken to evaluate the effects of the inclusion of linseed or demucilaged linseed in the diet on the performance, fat and fatty acid digestibilities, metabolisability of gross energy and intestinal microflora. 2. The dietary inclusions were 80 and 160 g/kg for linseed and 160 g/kg for demucilaged linseed. Diets were given to chickens from 1 to 23 d of age. 3. Incorporation of linseed in the diet, particularly at 160 g/kg, depressed weight gain and food utilisation. Digestibility of fat and single fatty acids and dietary metabolisable energy were reduced. 4. Inclusion of linseed in the diet markedly increased the viscosity of ileal digesta. Microbial activity in the ileum and caeca was not greatly affected, but there was a significant increase in the number of lactobacilli. 5. The antinutritional effects caused by feeding linseed were partially overcome by substituting demucilaged linseed for linseed in the diet. 6. Results from the current study suggest that the viscous properties of mucilage are a major factor in the observed antinutritional effects of linseed through increasing intestinal viscosity. Such effects might also be mediated by the gut microflora. PMID- 12737230 TI - The effect of short-term fasting on performance traits and rectal temperature of broilers during the summer season. AB - 1. The present study was conducted to determine if detrimental effects of high temperature on broiler performance could be overcome by short-term fasting and to examine the effect of food withdrawal on the responses of two commercial strains (S1 and S2). 2. An experimental design was used with three feeding groups and two strains. Birds in the control (C) group were fed ad libitum during the experimental period (0 to 6 weeks), while feed was withdrawn in the other groups between 10:00 and 16:00 h during weeks 5 and 6 (F1) or week 6 (F2), respectively. After standard brooding for the first 3 weeks temperature and humidity varied according to outside climate between 4 and 6 weeks of age. 3. Feeding strategy significantly affected body weight at week 6 (BW6) and daily weight gain during week 6 (DWG5-6). F2 gave the best results in BW6 and DWG5-6. Strain effect was significant on DWG4-5 and DWG5-6; S1 had higher DWG4-5 and S2 gained more at the last stage. S1C broilers had greater BW6 than S2C but the S2 strain gave a better response to short-term fasting compared with S1 and this resulted in significant strain by feeding interaction effect on BW6 and DWG5-6. 4. It was shown that short-term fasting was an effective method for controlling rectal temperature (RT) of broilers at 5 and 6 weeks of age. 5. Total food consumption (FCO-6), food conversion ratio (FCRO-6), mortality, and carcase characteristics were not affected by short-term fasting in the experimental conditions. 6. In conclusion, it can be recommended that broiler producers may practise 6 h of fasting during the hot period of the day without any adverse effect on slaughter weight and FCR when the birds are close to the average slaughter weight and a heat wave is expected. Strain differences in RT measurements also indicate the importance of choosing strains to be used in hot and warm climate conditions with attention to the thermoregulation ability of broilers. PMID- 12737229 TI - Effects of dietary protein concentration and specific amino acids on body weight, body composition and feather growth in young turkeys. AB - 1. Two randomised block factorial experiments were conducted to investigate the relationships between the effects of dietary crude protein and specific amino acid concentrations on the relative growth of the body and feathers of young turkeys. 2. Decreasing dietary crude protein concentration from 300 to 180 g/kg in experiment 1 reduced the body and breast muscle weights of a large male line of turkeys proportionally by 0.44 and 0.52 compared with 0.19 and 0.24 in a small traditional line. 3. Decreasing dietary crude protein concentration was associated with a maximum reduction in feather weight of 0.18 and 0.24 respectively in male line and traditional turkeys. The length of the feathers in the cranial region of the breast decreased from 26 to 19mm in the traditional line compared with an increase from 14 to 25 mm in male line turkeys. 4. Decreasing dietary crude protein concentration was associated with an increase in the fat content of the feather-free carcase. Male line turkeys had a higher carcase fat and lower feather dry matter content than the traditional turkeys. 5. It was concluded that dietary crude protein was preferentially partitioned to feather rather than muscle growth in the male line in contrast to a traditional line of turkeys in which the growth of feathers and muscle were affected equally. 6. In experiment 2, the amino acids arginine, valine, methionine and tyrosine were added separately to a common basal ration (180g CP/kg) to raise their concentration to that of the control ration (260 g CP/kg). Each ration was fed ad libitum to male line turkeys from 2 to 6 weeks of age. 7. Amino acid supplementation increased body and breast muscle weights. 8. Compared with the basal ration, tyrosine was associated with a reduction in feather weight whereas valine had no effect. Supplementation with arginine and methionine resulted in increased feather weights that were similar to that of the controls. 9. It was concluded that arginine and methionine were used preferentially and are essential for feather growth. Excess amino acids that are not required for feather growth such as tyrosine and valine were used for increased body growth and resulted in relatively poor feather cover. 10. The results suggest that feather growth was maintained as much as possible at the expense of body growth when the amino acid concentration of the ration was less than that required to maximise body and muscle weight gain in large male line turkeys. PMID- 12737231 TI - Relative bio-availability and utilisation of phosphatic fertilisers as sources of phosphorus in broilers and layers. AB - 1. Different concentrations of non-phytate phosphorus (NPP, 2.5, 3.0, 3.5, 4.0 and 4.5 g/kg diet) were given to broilers (8 to 42 d of age) to establish regressions between dietary NPP concentration and body weight gain and tibia ash content. Second and third experiments were conducted to study the feasibility of utilisation of different phosphatic fertilisers [ammonium phosphate (AP), ammonium polyphosphate (APP), single super phosphate (SSP), NPK (17:17:17, NPK) and NP (28:28:0, NPK)] in commercial broilers (8 to 42 d) and White Leghorn layers (252 to 364 d). 2. Phosphatic fertilisers were incorporated both in broiler (10 g calcium and 4.5 g NPP/kg) and layer (35 g calcium and 3.5 g NPP/kg) diets by replacing dicalcium phosphate (DCP) in toto. 3. The logarithmic curves obtained for predicting the body weight gain and tibia ash content at different levels of NPP used in experiment 1 were Y = 156.27 + 2,468.8 logX (r2= 0.958) and Y = 530.82 + 144.26 log X (r2 = 0.916), respectively. 4. Body weight gain and food intake in broilers given APP- or NP-supplemented diets were comparable to these in the DCP-fed group. Feeding of NPK, AP or SSP resulted in significant depression in weight gain and food intake and high excreta moisture content. Food/gain, Ca and P contents in tibia ash and serum were not influenced by the use of phosphatic fertilisers as P sources in broiler diets. 5. Tibia ash content in broilers fed on diets containing fertilisers was either similar to or significantly higher than that in the DCP-fed group. Broilers on AP or SSP retained more P and had higher tibia ash content than those on DCP. AP, SSP or NPK caused degenerative and necrotic changes in liver, kidney and intestine of broilers. 6. Relative bio-availability of P from APP or NP was better for body weight gain than AP, SSP or NPK, while the reverse was true for bone calcification. 7. APP and NP gave hen-d egg production similar to that of DCP-fed layers. Food intake was significantly reduced in layers fed on diets containing fertilisers. However, food/egg mass, egg weight and serum Ca and inorganic P contents were not influenced by inclusion of fertilisers in layer diets. 8. Except for AP, inclusion of fertilisers in layer diets reduced shell weight and shell thickness compared with the DCP-fed group. However, no apparent eggshell defects were found which could be attributable to diet. 9. Results of these experiments suggest that APP and NP can be used as the sole source of P both in broiler and layer diets, replacing DCP in toto. However, when utilising these P sources in layers, due attention should be given to shell quality. Fertilisers containing high F (AP and SSP) or K (NPK) reduced performance in broilers and layers and caused microscopic changes in liver, kidney and intestine in broilers. PMID- 12737232 TI - Interaction between genotype and dietary concentrations of methionine for immune function in commercial broilers. AB - 1. Growth, antibody response to sheep red blood cells (SRBC) and resistance to Escherichia coli were measured in broiler female chicks received from 4 (n = 100 in each) commercial genotypes (A, B, C and D) and fed with maize-soybean-deoiled rice bran based diets containing 4 concentrations of methionine (3.91, 4.46, 5.00 and 5.54 g/kg). The diets were fed ad libitum from 1 to 49 d of age. 2. Body weight gain and weight gain/food intake at 2 week intervals, response of broilers to inoculation of 0.5 ml of SRBC (0.5 or 2.5%), 0.1 ml of E. coli (10(-4) dilutions) culture, and 100 microg phytohaemogglutinin-P (PHA-P) at 43 d of age were measured. The responses to SRBC and E. coli inoculation were recorded at 5 d post inoculation (PI), while the responses to PHA-P were recorded at 12 and 24 h PI. 3. Genotype by methionine interaction was not significant for body weight gain, but significant differences in weight gain were observed among different genotypes. Variation in methionine concentration did not influence body weight gain or weight gain/food intake at 1 to 14, or 42 d of age. At 28 d of age, chicks fed on the 3.91 g methionine/kg diet weighted significantly less than those on the other methionine concentrations. Genotype by methionine interaction was observed for food efficiency at 0 to 28 d of age but not at other ages. 4. Antibody titres against SRBC and heart and air sac lesion score to E. coli challenge were not influenced by genotype-methionine interaction. Chicks given higher concentrations of methionine had higher antibody titres and greater cutaneous basophilic hypersensitivity (CBH) response than those given low levels of methionine. Also, variation was observed in expression of CBH response to PHA P among different genotypes. 5. It may be concluded that, although the commercial broiler chicks do not require more than 3.91 g methionine/kg for optimum growth and food efficiency, the immunity in terms of CBH response and antibody production to SRBC increased with the concentration of methionine in the diet in the majority of genotypes, indicating a higher methionine requirement for immunity than for weight gain. PMID- 12737233 TI - Effects of graded levels of Fusarium toxin-contaminated wheat and of a detoxifying agent in broiler diets on performance, nutrient digestibility and blood chemical parameters. AB - 1. A growth experiment was carried out with male broilers from d 1 to d 35 of age in order to evaluate the effects of the addition of a detoxifying agent (Mycofix Plus, Biomin GmbH, Herzogenburg, Austria) at different dietary proportions of wheat (0, 16.5, 33, 49.5 and 66%) contaminated with Fusarium mycotoxins (21.2 mg of deoxynivalenol and 406 microg of zearalenone, ZON, per kg of wheat) on growth performance, nutrient and zearalenone balance and clinical-chemical parameters. 2. An increase in dietary mycotoxin concentration resulted in a linearly related decrease in feed intake, a slight decrease in weight gain and an improvement in feed to gain ratio. 3. Apparent protein digestibility and net protein utilisation were higher in diets containing exclusively Fusarium toxin-contaminated wheat than control diets. 4. The proportions of beta-zearalenol, alpha-zearalenol and ZON of total ZON metabolites in excreta of broilers fed on the diets containing the Fusarium toxin-contaminated wheat were approximately 3, 21 and 76%. 5. Serum antibody titres to Newcastle disease virus decreased in a linear fashion with increasing mycotoxin concentration in the diets, whereas other clinical-chemical serum parameters (liver cell and muscle cell necrosis indicating enzymes, haemoglobin, haematocrit, magnesium, inorganic phosphate) were not influenced by increasing Fusarium toxin concentrations. 6. Supplementation of the diets with Mycofix Plus decreased performance in a manner independent of mycotoxin concentration. Moreover, some clinical-chemical serum parameters were significantly altered due to Mycofix Plus but also independently of the dietary mycotoxin concentration. PMID- 12737234 TI - Tea polyphenols reduce glucocorticoid-induced growth inhibition and oxidative stress in broiler chickens. AB - 1. The effects of dietary polyphenols (PP) on growth and oxidative stress in the corticosterone (CTC) treated broiler chickens model were studied. 2. Chicks (Cobb strain) were divided into 3 (CTC) x 3 (PP) blocks and given diets containing CTC at concentrations of 0, 10 and 20 mg/kg. 3. The body weight gain was lower when the birds were treated with CTC. However, the high dose of PP tended to reduce the effect of CTC. 4. The abdominal fat content, plasma triglyceride concentration and liver weight were increased by CTC and reduced by PP. 5. Muscle and liver thiobarbituric acid reactive substance (TBARS) were elevated by CTC and these effects were reduced by PP. Plasma CTC concentration was increased by dietary CTC treatment and decreased by PP. 6. In conclusion, our results indicate that PP can minimise growth inhibition, hyperlipidemia and oxidative stress induced by CTC treatment in broiler chickens. PMID- 12737236 TI - Effects of Lactobacillus cultures on growth performance, abdominal fat deposition, serum lipids and weight of organs of broiler chickens. AB - 1. The effects of a mixture of 12 Lactobacillus strains (LC) on the growth performance, abdominal fat deposition, serum lipids and weight of organs of broiler chickens were studied from 1 to 42 d of age. 2. One hundred and thirty six 1-d-old male broiler chicks were assigned at random to two dietary treatments: a basal diet (control), and a basal diet with 0.1% LC. 3. The supplementation of LC in broiler diets improved the body weight gain and feed conversion rate from 1 to 42 d of age and was effective in reducing abdominal fat deposition but only after 28 d of age. 4. The LC diets reduced serum total cholesterol, low density lipoprotein (LDL) cholesterol and triglycerides in broilers from 21 to 42 d of age. However, there was no significant difference in serum high density lipoprotein (HDL) cholesterol between control and LC-fed broilers. There was also no significant difference in the weights of organs of control and LC-fed broilers. 5. The results indicated that the mixture of 12 Lactobacillus strains have a hypolipidaemic effect on broilers. PMID- 12737235 TI - Effect of long-term heat exposure on peripheral concentrations of heat shock protein 70 (Hsp70) and hormones in laying hens with different genotypes. AB - 1. The effect of long-term, moderate heat stress (30 to 32 degrees C) on heat shock protein 70 (Hsp70) concentration in mononuclear blood cells and plasma concentrations of 3,5,3'-triiodothyronine (T3) and corticosterone in laying hens was investigated. 2. Three groups of 48 hens each (Ethopian line [Angete Melata, Na], New Hampshire [NH], F1 cross [Na x NH]) were divided into an experimental group (24 each) and a control group (24 each, ambient temperature 18 to 20 degrees C), respectively. All hens were kept in individual cages up to an age of 68 weeks and performance data were recorded. 3. Blood samples were taken from the wing vein of 12 hens from each group at weeks 22, 38, 51 and 65 (12 hens x 3 lines x 2 treatments). Mononuclear blood cells were isolated and Hsp70 concentrations were determined by Western Blot analysis with a monoclonal anti Hsp70 antibody. T3 and corticosterone were measured with commercially available ELISA and RIA kits, respectively. 4. The moderate heat stress caused significantly increased Hsp70 levels compared with the control groups in weeks 51 and 65. However, the responses of the lines were not uniform at different ages. 5. In contrast, T3 levels were significantly decreased in stressed birds regardless of line and age. There was no effect of treatment and line on corticosterone levels during the experimental period. 6. Our results indicate that Hsp70 and T3 levels are affected by mild heat stress applied over a long period but are both involved in independent mechanisms of acquisition of thermotolerance. Further investigations are necessary to clarify whether the observed differences in Hsp70 response between the genotypes are indicators for differences in thermotolerance. PMID- 12737237 TI - Distribution of N-acetylneuraminic acid and sialylglycan in eggs of the Silky fowl. AB - 1. The distribution of sialic acid in the eggs of original Silky fowl was investigated. The sialic acid contents of the yolk, albumen and the chalaza of a single egg were 205.2, 11.96 and 0.83 mg, respectively. 2. The sialic acid content of the yolk of Silky eggs was 11.5-fold higher than that of a conventional domestic fowl yolk. 3. Sialic acid isolated from Silky yolk was entirely N-acetylneuraminic acid (NeuAc). No N-glycolylneuraminic acid or O acetyl containing sialic acid was observed. 4. The structure of the major sialylglycan in Silky egg yolk was determined to be a disialyl-biantennary chain in which the NeuAc residues were alpha2-6 linked to glucose. No alpha2-3 linkage was observed. 5. Thus, the Silky fowl's egg provides an excellent source of NeuAc and sialylglycan. PMID- 12737239 TI - Marginal discrepancies and leakage of all-ceramic crowns: influence of luting agents and aging conditions. AB - PURPOSE: This study evaluated the marginal discrepancies and leakage of all ceramic crowns cemented with different luting agents after fatigue tests. MATERIALS AND METHODS: Forty-eight all-ceramic crowns were cemented onto natural molars. Zinc-phosphate cement, compomer cement, and an adhesive composite resin luting system were used in 16 specimens each. Sixteen metal-ceramic crowns were cemented with zinc-phosphate cement as a control. Half of the specimens in each group were fatigued in a chewing simulator for 600,000 loading cycles with 3,500 thermocycles. The others received the 3,500 thermocycles only. An impression replica technique and SEM were used for evaluation of the marginal discrepancies. Leakage of the specimens was microscopically assessed. RESULTS: There were no significant differences between marginal discrepancies of the groups, with the exception of porcelain shoulder margins in the metal-ceramic group, which had significantly larger discrepancies (P < .01). There was no significant effect of the fatigue tests on marginal discrepancies. However, significant differences of leakage were found between the groups cemented with the different luting agents after both fatigue tests. The adhesive composite resin luting system demonstrated the least leakage. The compomer cement showed an intermediate level of leakage, and the zinc-phosphate cement showed severe leakage that extended through the dentinal tubules to the pulp chambers. No significantly different effect of the two fatigue tests on the leakage was found. CONCLUSION: The adhesive composite resin luting system showed clinically acceptable marginal discrepancies and an excellent ability to minimize leakage of all-ceramic crowns. PMID- 12737238 TI - Beta2-agonists and cAMP inhibit protein degradation in isolated chick (Gallus domesticus) skeletal muscle. AB - 1. The role of beta2-agonist and of cAMP in chick skeletal muscle proteolytic pathways and protein synthesis was investigated using an in vitro preparation that maintains tissue glycogen stores and metabolic activity for several hours. 2. In extensor digitorum longus (EDL) muscle total proteolysis decreased by 15 to 20% in the presence of equimolar concentrations of epinephrine, clenbuterol, a selective hbetaagonist, or dibutyryl-cAMP. Rates of protein synthesis were not altered by clenbuterol or dibutyryl-cAMP. 3. The decrease in the rate of total protein degradation induced by 10(-5)M clenbuterol was paralleled by a 44% reduction in Ca2+-dependent proteolysis, which was prevented by 10(-5)M ICI 118.551, a selective fbeta2antagonist. 4. No change was observed in the activity of the lysosomal, ATP-dependent, and ATP-independent proteolytic systems. Ca2+ dependent proteolytic activity was also reduced by 58% in the presence of 10(-4)M dibutyryl-cAMP or isobutylmethylxanthine. 5. The data suggest that catecholamines exert an inhibitory control of Ca2+-dependent proteolysis in chick skeletal muscle, probably mediated by fbeta2adrenoceptors, with the participation of a cAMP-dependent pathway. PMID- 12737240 TI - Comparing the efficacy of mandibular implant-retained overdentures and conventional dentures among middle-aged edentulous patients: satisfaction and functional assessment. AB - PURPOSE: The aim of this randomized clinical trial was to compare the relative efficacy of mandibular overdentures retained by only two implants and a bar attachment with conventional dentures. MATERIALS AND METHODS: Edentulous adults, aged 35 to 65 years, were randomly assigned to two groups that received either a mandibular conventional denture (n = 48) or an overdenture supported by two endosseous implants with a connecting bar (n = 54). All subjects rated their general satisfaction and other features of their original dentures and their new prostheses (comfort, stability, ability to chew, speech, esthetics, and cleaning ability) on 100-mm visual analogue scales prior to treatment and 2 months postdelivery. Oral health-related quality of life was also evaluated pre- and posttreatment. RESULTS: Multiple regression analysis revealed that the mean general satisfaction was significantly higher in the overdenture group than in the conventional denture group (P = .0001). Age, gender, marital status, and income were not significantly associated with ratings of general satisfaction. Furthermore, the implant group gave significantly higher ratings on three additional measures of the prostheses (comfort, stability, and ease of chewing; P < .05). CONCLUSION: A mandibular two-implant overdenture opposed by a maxillary conventional denture is a more satisfactory treatment than conventional dentures for edentulous middle-aged adults. PMID- 12737241 TI - Systematic review of 10 years of systematic reviews in prosthodontics. AB - PURPOSE: The objective was to make an inventory of systematic reviews in the field of prosthodontics and to assess the strength of evidence yielded by these studies. MATERIALS AND METHODS: The literature was searched using MEDLINE (keywords "dental" in subset combined with "meta-analysis" in publication type, and "dental" in subset combined with "systematic review"). Reviews related to prosthodontics were selected by hand. Analogies between the reviewing processes were assessed, and the quality was described. RESULTS: There were 138 articles qualifying as either systematic reviews or meta-analyses. Of these, 13 reported pooled data on prosthodontic subjects. Two pairs of reviews were identified as dealing with comparable items; the others described all different subjects. In one pair, the studies reviewed the survival of conventional fixed partial dentures (FPD); the other pair was on single-tooth implants. The pooled results within each pair were almost equal. For the FPD reviews, 65% of the unity of studies was included in both reviews. For the single-tooth implants, 29% of the potentially useful studies were included in both reviews. The data pooling processes showed the same pattern. One large study included in both reviews explained a large part of the similarity of the combined survivals of FPDs. For the single-tooth implant reviews, the largest common study explained 20% of the similarity. CONCLUSION: Although there were methodologic differences between the paired reviews, they produced similar results. The outcomes of the evaluated reviews may be used as prognostic data; however, they cannot be used for direct comparison of treatments. PMID- 12737242 TI - In vitro study of a mandibular implant overdenture retained with ball, magnet, or bar attachments: comparison of load transfer and denture stability. AB - PURPOSE: When implants are used to support a removable overdenture, the optimal stress distribution to minimize both forces on the implants and denture movement is desirable. This study compared the stress patterns generated around implants and denture movement among three retention systems. MATERIALS AND METHODS: Two root-form implants were anchored in a mandibular model made of resin, and a removable overdenture on which all experiments were performed was fabricated. The surface of the model was covered with a layer of impression material to simulate oral mucosa. Ball/ O-ring, bar/clip, and magnetic attachments were used. A vertical force was applied to the left first molar and gradually increased from 0 to 50 N in 5-N steps. The resultant stress distribution and denture movement were evaluated. RESULTS: The ball/O-ring attachment transferred the least stress to both implants and produced less bending moment than the bar/clip attachment. Vertical force applied to the bar/clip attachment created immediate stress patterns of greater magnitude and concentration on both implants. CONCLUSION: This in vitro study suggested that the use of the ball/O-ring attachment could be advantageous for implant-supported overdentures with regard to optimizing stress and minimizing denture movement. PMID- 12737243 TI - Fracture toughness (K(IC) of a hot-pressed core ceramic based on fractographic analysis of fractured ceramic FPDs. AB - PURPOSE: The objective of this study was to test the hypothesis that there is no significant difference between the fracture toughness (K(IC)) of an experimental hot-pressed core ceramic measured by fractographic analysis of failed ceramic prostheses and the values determined by other standard methods. MATERIALS AND METHODS: Four groups were subjected to one of four test methods: group 1 = indentation strength technique (standard numerical calculation); group 2 = indentation strength technique (fractographic analysis); group 3 = flexure test of precracked specimens (fractographic analysis); and group 4 = fractographic analysis of failed three-unit fixed partial dentures (FPD). For groups 1 to 3, 20 ceramic bar specimens were subjected to three-point flexure at a cross-head speed of 0.5 mm/min until fracture occurred. For group 4, 10 failed FPDs were collected from a previous study. Stress values at failure were calculated from either a flexure stress equation (groups 1 to 3) or from finite element analyses (group 4). K(IC) values were calculated from an equation and fractographic measurement data. RESULTS: Mean fracture toughness ranged from 3.1 MPa x m1/2 (SD 0.2) (group 1) to 3.4 MPa x m1/2 (SD 0.2) (group 4). The mean K(IC) value for group 1 was significantly different from that of group 4; however, no significant differences were found between groups 1, 2, and 3, or between groups 2, 3, and 4. CONCLUSION: Fracture toughness for an experimental hot-pressed core ceramic measured by fractographic analysis in combination with finite element analysis was comparable with the values determined with other standard fractographic methods. PMID- 12737245 TI - Bond strength between polymer resin-based cement and porcelain-dentin surfaces: influence of polymerization mode and early cyclic loading. AB - PURPOSE: The objective of this study was to verify the influence of resin cement polymerization mode and early cyclic loading on the bond strength of porcelain dentin specimens. MATERIALS AND METHODS: Two cementation systems, Choice/One-Step and RelyX ARC/Single Bond, were tested. Porcelain truncated cones were bonded into cavities prepared in sections of bovine root dentin. Each system was tested with the resin cement in dual-cure and self-cure mode (with and without photoactivation, respectively). Half of the specimens were load cycled 15 minutes after cement mixing (60 N, 20 times). The other half were not submitted to mechanical loading (control). After 24-hour storage in distilled water (37 degrees C), the extrusion shear test was performed. Data were analyzed by three way ANOVA. RESULTS: With both systems, some specimens built with the cement in self-cure mode debonded during cyclic loading (RelyX ARC/Single Bond 47%; Choice/One-Step 13%). No significant system effects were detected. Mean bond strength under the dual-cure mode (13.4 MPa, SD 3.6) was significantly higher than the corresponding mean for the self-cured specimens (5.7 MPa, SD 2.6). No reduction in bond strength was observed for the specimens that survived the early cyclic loading. CONCLUSION: Resin cements in dual-cure mode presented higher bond strengths than self-cured materials. Early load cycling caused debonding of some self-cured specimens. However, the bond strength of the survivor specimens was not affected by load cycling. PMID- 12737244 TI - Cytotoxicity of antimicrobial tissue conditioners containing silver-zeolite. AB - PURPOSE: The purpose of this study was to determine the effects of incorporating antimicrobial silver-zeolite on the in vitro cytotoxicities of five tissue conditioners against the living dermal model, which consisted of normal human dermal fibroblasts in a collagen lattice. MATERIALS AND METHODS: Samples of each tissue conditioner containing silver-zeolite (2 wt% and 5 wt%) were prepared; samples without silver-zeolite were used as a control. Cytotoxicity of each sample against the living dermal model was evaluated by cell viability with MTT assay. RESULTS: Cell viabilities for Visco-Gel, GC Soft-Liner, FITT, and SR Ivoseal decreased with increasing silver-zeolite content. Cell viability for Shofu Tissue Conditioner showed no significant difference with degree of silver zeolite incorporation and had higher values than other tissue conditioners. CONCLUSION: Individual compounds eluted from tissue conditioners and silver ions released from silver-zeolite might cause such cytotoxicity. The results suggest that Shofu Tissue Conditioner is a possible candidate for a novel antimicrobial tissue conditioner containing silver-zeolite because it showed the highest cell viability and the smallest influence of incorporating silver-zeolite. PMID- 12737247 TI - Pulmonary risk of intraoral surface conditioning using crystalline silica. AB - PURPOSE: This study evaluated the pulmonary risk caused by possible respirable dust of Al2O3 and SiO(x) resulting from chairside tribochemical sandblasting procedures in a dental office. MATERIALS AND METHODS: Dust was collected using a trap near the working field, and quantitative morphologic determination and identification were performed with SEM and EDAX. Forty blasting processes (total time 20 minutes) were aimed at a dummy to obtain maximum pollution of the workplace. Respirable dust fraction was measured using personal air samplers with an 8-microm cellulose-nitrate filter and a volume flow rate of 2 L/min. Mass of the respirable dust fraction was determined, and respirable free crystalline silica was identified with the help of infrared spectroscopy. RESULTS: Blasting of metal or ceramic surfaces with tribochemical agents produces respirable and potentially harmful SiO(x) and Al2O3 particles with a diameter of less than 5 microm, showing a total concentration in the air of less than 0.3 mg/m3. With and without dental suction, the concentration of the respirable free crystalline silica was smaller than 0.02 mg/m3. CONCLUSION: Concern regarding the risk of chairside tribochemical methods and possible impairment of health of patients and dental staff is unfounded, even under extreme conditions or without protective measures, since the concentrations of SiO(x) found in the air of the workplace were far below the current threshold value of 0.15 mg/m3. PMID- 12737246 TI - A long-term retrospective and clinical follow-up study of In-Ceram Alumina FPDs. AB - PURPOSE: The purpose of this study was to evaluate the long-term outcome of In Ceram Alumina fixed partial dentures (FPD) performed in a general dental practice from 1992 to 1996. MATERIALS AND METHODS: The study was conducted as a retrospective assessment of up to 9 years of patient records and a clinical follow-up examination of patients treated with In-Ceram Alumina FPDs. In 37 patients, 42 FPDs had been inserted during the selected period. After randomized selection, 16 patients with 18 FPDs were examined clinically. The most common restorations comprised two and three units. Cantilever extensions were present on 64% of the FPDs. Sixty-two percent of the FPDs extended into the posterior region. RESULTS: The mean time in function for the 42 FPDs was 76 months (range 2 to 110 months), with 86% being followed for > 5 years. No adverse effects to either periodontal or pulpal tissues were recorded. The technical quality was very good, and patient satisfaction very high. Five FPDs fractured during the observation period, resulting in a total failure rate of 12%. Two of these FPDs fractured as a consequence of external trauma. Excluding these, the total survival rate during the observation period was 93%. Cumulative survival rate according to life table analysis was 93% after 5 years and 83% after 10 years. CONCLUSION: The results suggest that the In-Ceram Alumina short-span FPD is a viable prosthetic alternative. PMID- 12737248 TI - Strength and mode of failure of unidirectional and bidirectional glass fiber reinforced composite materials. AB - PURPOSE: The flexural strength of two commercially available dental fiber reinforced composites (FRC) (glass fiber-reinforced composite material), one unidirectional and the other bidirectional, were investigated. MATERIALS AND METHODS: Ten uniform beams of core materials and ten beams of laminated core materials were fabricated for FibreKor and Vectris Frame. The specimens were subjected to a three-point bending test. Flexural strength for both FRC materials was determined with and without their composite laminations. The strength data were analyzed using the Weibull method. Modes of failure for both systems were determined using SEM. RESULTS: The strength of FibreKor was significantly greater than that of Vectris Frame when comparing the core materials with and without their respective composite laminations. Mode of failure of FibreKor was predominantly debonding with fiber fracture. Vectris Frame did not exhibit debonding failure. Mode of failure for Vectris Frame was fiber fracture with delamination and matrix microfracture. CONCLUSION: FibreKor, a unidirectional FRC, demonstrated higher flexural strength than Vectris Frame, a bidirectional FRC. Debonding of fibers from the matrix possibly contributed to toughening mechanisms such as crack deflection, fiber pullout, and fiber bridging in FibreKor. PMID- 12737249 TI - Fracture strength and failure mode of five different single-tooth implant abutment combinations. AB - PURPOSE: The purpose of this study was to evaluate the fracture strength and mode of failure of five different single-tooth abutment-implant combinations before and after cyclic loading in the artificial mouth. MATERIALS AND METHODS: Eighty standardized maxillary central incisor crowns were fabricated for five test groups with 16 specimens each (group 1: Steri-Oss/Novostil; group 2: Steri Oss/Anatomic abutment; group 3: Steri-Oss/straight HL; group 4: IMZ Twin +/Esthetic abutment; group 5: Osseotite/gold UCLA). Specimens were adhesively luted using Panavia 21, and half were artificially aged via chewing simulation and thermocycling. All nonaged and the surviving aged specimens were tested for fracture strength resistance using compressive load on the palatal surfaces of the crowns. RESULTS: After exposure to the artificial oral environment, survival was as follows: six in group 1; eight in groups 2, 3, and 5; and seven in group 4. Median fracture strengths before and after loading in the artificial mouth were 537 N, 817 N, 893 N, 473 N, and 743 N for groups 1 to 5, respectively. There were no statistically significant differences in fracture strength before and after exposure to the artificial mouth. There were significant differences between the implant systems in both stages of the experiment. In all five groups, abutment-screw bending and abutment fractures (except group 1) were present after static loading, and all implant necks in group 4 showed distortions. CONCLUSION: The artificial mouth is a useful tool to check the implant-abutment-screw interface stability. The physical properties of screws and screw joints of groups 1 and 4 have to be improved. Groups 2, 3, and 5 have the potential to withstand physiologic biting forces. PMID- 12737250 TI - Implant-supported fixed partial prostheses: a retrospective study. AB - PURPOSE: The purpose of this retrospective study was to present treatment outcome and patient reactions to rehabilitation with implant-supported fixed partial prostheses. MATERIALS AND METHODS: Eighty-three patients were consecutively treated with implant-supported fixed partial prostheses (Branemark system) from 1986 to 1995. Seventy-six of these 83 patients were examined (66 maxillary and 31 mandibular prostheses). The mean observation time was 53.9 months. In total, 285 implants were placed. Eleven implants were lost before loading. The first 41 prostheses were removed and the implants examined regarding the criteria for "success" and tightness of the screw joints. Only one implant had lost integration. RESULTS: The survival rate before and after loading was 96%, which included implants placed in augmented bone. All prostheses were stable at the time of examination. In prostheses with cantilevers (98 implants), 12% of the gold screws and 17% of the abutment screws showed a "not acceptable" loosening, compared to none in the prostheses without cantilevers (17 implants). The difference was not statistically significant. The mean marginal bone loss was 0.4 mm for the first year after prosthesis insertion and less than 0.1 mm per year in the following years. The most frequent prosthesis design was one pontic supported by two implants. Prostheses made in gold acrylic and titanium acrylic had more complications and showed a higher need for repair than metal-ceramic restorations. Patients reacted very positively to the esthetic results and comfort with eating, and were overall satisfied with their prostheses. CONCLUSION: Implant-supported fixed partial prostheses seem to have a very good prognosis and are well-accepted by patients. PMID- 12737252 TI - Effect of clenching level on mandibular displacement in Kennedy Class II partially edentulous patients. AB - PURPOSE: Forceful clenching in the intercuspal position has the potential to cause significant mandibular displacement. Such a displacement can be expected to be exaggerated in patients without molar support. The appropriate clenching level for intercuspal position registration or evaluation in these patients has never been clarified. The aim of this study was to clarify the effect of clenching level and absence of posterior occlusal support on mandibular displacement. MATERIALS AND METHODS: Thirteen women with a unilateral edentulous area posterior to the first premolar were enrolled in this study. They were asked to perform intercuspal clenching at various occlusal force levels, and 3-D mandibular displacements were measured by an optoelectronic method. The effects of "clenching force level" and "side of molar tooth loss" on the displacements of the condylar and second molar points were analyzed. RESULTS: The mandible was elevated without substantial horizontal displacements during each clenching task. The effect of clenching level on vertical mandibular displacement was significant, with a clenching level > or = 50% of maximal voluntary contraction causing significantly greater elevations (P < .05). Clenching > or = 50% of maximal voluntary contraction caused consistent tipping of the mandible, with greater elevation on the edentulous side (P < .05). CONCLUSION: The clenching level should not surpass 50% of maximal voluntary contraction for registration or evaluation of intercuspal position in Kennedy Class II patients. PMID- 12737251 TI - An up to 15-year longitudinal study of 515 metal-ceramic FPDs: Part 2. Modes of failure and influence of various clinical characteristics. AB - PURPOSE: This study reported on the modes of failure and the influence of various clinical characteristics on the outcome of 515 metal-ceramic fixed partial dentures (FPD) involving 1,209 abutment and 885 pontics placed by one operator in a specialist prosthodontic practice between January 1984 and December 1997. MATERIALS AND METHODS: Fifty-three percent of maxillary and 84% of mandibular FPDs involved one or more nonrigid abutment-pontic connections. Patients were recalled in 1993 (review 1) and 1998 (review 2) and clinically examined by the author. The modes of failure and influence of clinical characteristics, including abutment-pontic connection, post design in nonvital abutments, and regularity of professional maintenance, were evaluated. RESULTS: At review 2, 80% of the FPDs were still in function, while 9% required retreatment. Tooth fracture accounted for 38%, caries for 11%, loss of retention for 13%, and periodontal breakdown for 27% of retreatments. Nonrigid abutment-pontic connection in posterior FPDs and regular professional maintenance were associated with significantly reduced failure rates. Except in posterior molars, there was no difference in failure rates between parallel-sided preformed stainless steel serrated posts and cast gold alloy posts. CONCLUSION: Tooth fracture was the most common reason for retreatment of the FPDs, although the rate of periodontal breakdown and caries increased significantly with time. Nonrigid abutment-pontic connection and regular professional maintenance were associated with significantly reduced failure rates. Post design and composition were not related to outcome. PMID- 12737253 TI - Retention, marginal leakage, and cement solubility of provisional crowns cemented with temporary cement containing stannous fluoride. AB - PURPOSE: This in vitro study investigated the (1) retention and microleakage of provisional crowns cemented with temporary cements to which stannous fluoride (SnF2) was added, and (2) solubility of these cements. MATERIALS AND METHODS: Provisional crowns were constructed of acrylic resin with shoulder preparations for 12 molars. The crowns were luted with Tempbond, Tempbond NE, and Freegenol temporary cements, and also with SnF2 added to these cements. Specimens were thermocycled 100 times, stored for 6 days, and immersed in 0.5% basic fuschin. Seven days after cementation, crown removal (retention) tests were conducted. Marginal leakage was assessed using a five-level scale to score dye penetration. Solubility in water of the cements with and without SnF2 was assessed using cement disks. RESULTS: Freegenol was more retentive than the other cements. The incorporation of SnF2 significantly increased the retention capacity of Freegenol and Tempbond NE but had no effect on Tempbond. Tempbond showed significantly higher dye penetration than Freegenol. The addition of SnF2 did not alter the dye penetration of the cements. There were no significant differences in the solubility of the cements. However, the incorporation of SnF2 increased the solubility of Freegenol and Tempbond NE (P < .001) and Tempbond (P < .01). CONCLUSION: The addition of SnF2 increased the retention of temporary crowns cemented with Tempbond NE and Freegenol but did not affect the retention of those cemented with Tempbond. The marginal leakage of crowns cemented with the tested temporary cements with and without the incorporation of SnF2 was similar. However, the addition of SnF2 increased the solubility of the cements. PMID- 12737254 TI - Comparisons of precision of fit between cast and CNC-milled titanium implant frameworks for the edentulous mandible. AB - PURPOSE: The purpose of this study was to investigate and compare the precision of fabrication in repeatqdly produced computer numeric controlled (CNC)-milled frameworks with consventional castings, and to analyze the distortion from application of different veneering materials. MATERIALS AND METHODS: Twenty identical titanium frameworks were fabricated by means of a CNC milling technique for the same master model. Five conventional frameworks were cast as a control group to the same model. The frames were measured with regard to fit in a coordinate measuring machine linked to a computer. Measurements were made during different stages of handling of the titanium framework, and after veneering materials had been applied. RESULTS: The CNC frameworks showed a statistically better fit and precision of fabrication compared to conventional castings (P < .05). The application of veneering material did not statistically affect the fit of the titanium frameworks (P > .05). CONCLUSION: It is possible to fabricate implant-supported frameworks by means of the present CNC technique with a very high precision and repeatability. PMID- 12737255 TI - Complications after treatment with implant-supported fixed prostheses: a retrospective study. AB - PURPOSE: The aim was to retrospectively evaluate the frequencies of different complications, as well as the number of visits to dentists because of such complications, after treatment with implant-supported fixed prostheses. MATERIALS AND METHODS: The study group comprised 75 patients who had been treated with implant-supported fixed prostheses 3 years earlier. All case records were scrutinized, and notes of complications in association with implants and superstructures were registered. RESULTS: The most common intervention made was occlusal adjustment/selective grinding of the prostheses. Complications in association with both implants and superstructures were fairly common. The most frequent complication was fractures of the acrylic resin matrix, including artificial acrylic resin teeth. Consultations because of periimplant mucosal inflammation were much more common among women compared to men, while complications that could be attributed to heavy loading tended to be more common in men. CONCLUSION: Complications with both implants and superstructures are fairly common after treatment with implant-supported fixed prostheses. Regular follow-ups to maintain optimal function in these patients are thus mandatory. PMID- 12737257 TI - Change in acceptability of barley plants to aphids after exposure to allelochemicals from couch-grass (Elytrigia repens). AB - The response of the bird cherry-oat aphid, Rhopalosiphum padi, to barley plants was investigated following exposure of the plants to root allelochemicals from the aggressive weed couch-grass, Elytrigia (Agropyron) repens. Plants were treated either with root exudates from living couch-grass plants or with previously identified couch-grass root compounds 15-hydroxyindole-3-acetic acid, DL-5-hydroxytryptophan, L-5-hydroxytryptophan hydrate, and 6-hydroxy-1,2,3,4 tetrahydro-beta-carboline-3-carboxylic acid (carboline)] either separately or in mixtures. In choice and no-choice settling tests, aphid acceptance of barley plants was significantly reduced following treatment with root exudates, and the carboline when tested alone or in combination with the other compounds. In contrast, the other compounds without the carboline were less active in reducing aphid acceptance. In a probing bioassay, individual substances were either neutral or stimulatory to aphids, indicating that the reduced settling was probably not due to direct effects on aphids, but rather due to effects on the plant. This was confirmed in olfactometer assays, in which aphids were repelled by odors from barley plants following treatment with a mixture containing all four chemicals. PMID- 12737256 TI - Consensus statement on fiber-reinforced polymers: current status, future directions, and how they can be used to enhance dental care. AB - The Second Scientific Fibre Reinforcement Symposium was held in Nijmegen, The Netherlands, on October 13th, 2001. The participants were invited speakers with extensive scientific and clinical backgrounds in glass fiber and polyethylene fiber research. The symposium reports focused on four areas of fiber reinforcement research: materials development, laboratory testing, clinical systems development, and clinical data. The consensus reached on the current status and future directions of this technology is reported here. PMID- 12737258 TI - First isolation of natural cyanamide as a possible allelochemical from hairy vetch Vicia villosa. AB - Cyanamide was isolated from the leaves and stems of hairy vetch (Vicia villosa), guided by plant growth inhibitory activity against lettuce (Lectuca sativa) seedlings. A large proportion of the inhibitory activity in the crude extract was explained by the presence of cyanamide, suggesting it to be a possible allelochemical in this species. The amount in a 9-day-old seedling, which had been grown without nutrients, reached approx. 40 times that of a nongerminated seed, demonstrating cyanamide biosynthesis in the seedlings. This is the first report on the isolation of a possible allelochemical from hairy vetch and also of the finding of cyanamide as a natural product. PMID- 12737259 TI - Genetic variation and relationships of constitutive and herbivore-induced glucosinolates, trypsin inhibitors, and herbivore resistance in Brassica rapa. AB - We examined genetic variation in inducibility and in constitutive and herbivore induced levels of glucosinolates, trypsin inhibitors, and resistance to herbivory in families of Brassica rapa originating from a wild population. We also examined phenotypic and genetic correlations among absolute levels of these traits in control and induced plants. We grew seedlings of 10 half-sib families in pairs in pots, and exposed one plant per pair to folivory by Trichoplusia ni larvae. Two days later, we sampled all plants for total glucosinolate and trypsin inhibitor levels and examined the preference and consumption by T. ni larvae of previously damaged (induced) and undamaged (control) plants. There was no significant variation among sire families in the induction of glucosinolates or trypsin inhibitors by T. ni feeding. Total glucosinolate levels in either control or induced plants did not vary by family. In contrast, trypsin inhibitor levels in both control and induced plants varied significantly by family. Trichoplusia ni fed less on induced plants than on control plants in the bioassay, but neither the induction of resistance by prior T. ni feeding nor absolute levels of damage done to control and induced plants varied significantly by sire family. Temporal blocking strongly affected trypsin inhibitor levels and the response of some families in the bioassays. There were no significant phenotypic or genetic correlations of levels of glucosinolates or trypsin inhibitors with each other or with damage in either control or induced plants. Overall, these results suggest that in the B. rapa population that we studied, both total glucosinolate content and biological resistance to herbivory by T. ni was nonvariable and almost universally inducible by prior T. ni feeding. In contrast, control and induced levels of trypsin inhibitors varied genetically and have the capacity to respond to future selection imposed by herbivores. However, the role of these defenses in constitutive or induced resistance to T. ni in this species remains unclear. PMID- 12737260 TI - Herbivore-induced responses in alfalfa (Medicago sativa). AB - The herbivore-induced response of alfalfa (Medicago sativa) was examined through assays with Spodoptera littoralis larvae and analyses of important secondary substances. In food preference experiments, larvae preferred young undamaged alfalfa plants over plants that had been damaged by feeding larvae 5 and 7 days earlier. while no difference in feeding preferences could be detected 1. 9. and 14 days after damage. This suggests a peak in the herbivore induced resistance of alfalfa approximately one week after initial damage. The induced resistance in young plants was also shown to be systemic, while older flowering plants failed to show increased resistance after defoliation. Larvae gained weight slower and had lower pupal mass when fed damaged alfalfa than when fed undamaged alfalfa. Levels of total saponins were increased in foliage of damaged alfalfa, and detailed analyses of specific saponin components revealed doubled concentrations of 3GlcA,28AraRhaXyl medicagenate (medicagenic acid bidesmoside) and 3GlcAGalRha soyasapogenol B (soyasaponin I). Levels of the flavonoid apigenin (as free aglycone) also were increased in herbivore damaged plants. The herbivore-induced response of alfalfa was significantly weaker than that of cotton: S. littoralis larvae given a choice of undamaged cotton and undamaged alfalfa preferred to feed on cotton, whereas preferences shifted towards alfalfa when plants were damaged. PMID- 12737261 TI - Identification of host fruit volatiles from hawthorn (Crataegus spp.) attractive to hawthorn-origin Rhagoletis pomonella flies. AB - Solid-phase microextraction (SPME) and gas chromatography coupled with electroantennographic detection (GC-EAD) were used to identify volatile compounds from hawthorn fruit (Crataegus spp.) acting as behavioral attractants for hawthorn-infesting Rhagoletis pomonella flies. Consistent EAD activity was obtained for six chemicals: ethyl acetate (94.3%), 3-methylbutan-1-ol (4.0%), isoamyl acetate (1.5%), 4,8-dimethyl-1,3(E),7-nonatriene (0.07%), butyl hexanoate (0.01%), and dihydro-beta-ionone (0.10%). In a flight-tunnel bioassay, there was a dose-related increase in the percentage of flies flying upwind to the six component mixture. Hawthorn-origin flies also made equivalent levels of upwind flight with the synthetic blend and an adsorbent extract of volatiles collected from whole fruit, each containing the same amount of the 3-methylbutan-1-ol compound. Significantly lower levels of upwind flight occurred to a previously identified volatile blend of ester compounds that attracts R. pomonella flies infesting domestic apples, compared with the hawthorn volatile mix. Selected subtraction assays showed further that the four-component mixture of 3 methylbutan-1-ol, 4,8-dimethyl-1,3(E),7-nonatriene, butyl hexanoate, and dihydro beta-ionone also elicited levels of upwind flight equivalent to the six-component mix. Removal of 3-methylbutan-1-ol from the four-component blend resulted in complete loss of upwind flight behavior. Removal of dihydro-beta-ionone, 4,8 dimethyl-1,3(E),7-nonatriene, or butyl hexanoate from the four-component mixture resulted in significant decreases in the mean number of upwind flights compared to the four- or six-component mixtures. PMID- 12737262 TI - Sequestration and metabolism of protoxic pyrrolizidine alkaloids by larvae of the leaf beetle Platyphora boucardi and their transfer via pupae into defensive secretions of adults. AB - Several neotropical leaf-beetles of the genus Platyphora ingest and specifically metabolize plant acquired pyrrolizidine alkaloids (PAs) of the lycopsamine type (e.g., rinderine or intermedine) and enrich the processed alkaloids in their exocrine defensive secretions. In contrast to the related palaearctic leaf beetles of the genus Oreina, which absorb and store only the non-toxic alkaloid N oxides, Platyphora sequesters PAs exclusively as protoxic tertiary amines. In this study, the ability of P. boucardi larvae to accumulate PAs was investigated. Tracer studies with [14C]rinderine and its N-oxide revealed that P. boucardi larvae, like adult beetles, utilize the two alkaloidal forms with the same efficiency, but accumulate the alkaloid as a tertiary amine exclusively. Ingested rinderine is rapidly epimerized to intermedine, which is localized in the hemolymph and all other tissues; it is also detected on the larval surface. Like adults, larvae are able to synthesize their own alkaloid esters (beetle PAs) from orally administered [14C]retronecine and endogenous aliphatic 2-hydroxy acids. These retronecine esters show the same tissue distribution as intermedine. A long term feeding experiment lasting for almost four months revealed that retronecine esters synthesized from [14C]retronecine in the larvae are transferred from larvae via pupae into the exocrine glands of adult beetles. Pupae contain ca. 45% of the labeled retronecine originally ingested, metabolized, and stored by larvae; ca. 12% of larval radioactivity could be recovered from the defensive secretions of adults sampled successively over two and a half months. Almost all of this radioactivity is found in the insect-made retronecine esters that are highly enriched in the defensive secretions, i.e., more than 200-fold higher concentration compared to pupae. PMID- 12737263 TI - Effect of conspecific and heterospecific feces on foraging and oviposition of two predatory ladybirds: role of fecal cues in predator avoidance. AB - Growing evidence suggests a flow of chemical information from higher to lower trophic levels that affects foraging and oviposition of 'prey' in response to potential risks from predators. This was investigated in two species of ladybird predators of aphid, Harmonia axyridis and Propylea japonica. H. axyridis is known to be the stronger intraguild predator and P. japonica to be the more frequent intraguild prey in interactions of these two species. These ladybirds share aphid prey on mugworts, hibiscus, and Italian ryegrasses in fields of northern Japan but largely avoid each other on the same plant. Fecal cues of these ladybird predators were found to contribute in their assessment of predation risk from conspecific and heterospecific competitors in common habitats. Gravid females of H. axyridis reduced rates of feeding and oviposition when exposed to feces of conspecifics, but not when exposed to feces of P. japonica. In contrast, gravid females of P. japonica reduced feeding and oviposition when exposed to feces of both H. axyridis and its own species. Females of both ladybird species exhibited similar behavior in response to water extracts of feces. For P. japonica, the influence of heterospecific feces was greater than that of conspecific feces. Our results demonstrate that feces of ladybirds contain odors that have the potential to deter the feeding and oviposition activities of conspecific as well as heterospecific ladybirds. Such deterrence allows these insects to avoid predation risk. Differences in responses of the two predators are discussed. PMID- 12737265 TI - Sex pheromone of horse-chestnut leafminer Camneraria ohridella and its use in a pheromone-based monitoring system. AB - Gas chromatography combined with electroantennographic detection (GC-EAD), electroantennography (EAG), and wind-tunnel and field experiments were used to reinvestigate the composition of Cameraria ohridella (Lepidoptera, Gracillariidae, Lithocolletinae) sex pheromone. The GC-EAD experiments showed one EAD-active area corresponding to the major pheromone component. (8E,10Z) tetradeca-8,10-dienal. The EAG experiments proved that (9E)-tetracedecenal and stereoisomers of (8E,10Z)-tetradeca-8,10-dienal exhibited significant electrophysiological activity and could, therefore, be considered as possible minor pheromone components. However, wind-tunnel and field experiments demonstrated that none of these compounds affect the efficacy of the main pheromone component. A monitoring system based on (8E,10Z)tetradeca-8,10-dienal was developed and used to study the flight activity of C. ohridella. PMID- 12737266 TI - Presence of long-lasting peripheral adaptation in oblique-banded leafroller, Choristoneura rosaceana and absence of such adaptation in redbanded leafroller, Argyrotaenia velutinana. AB - Pre-exposure of male oblique-banded leafrollers, Choristoneura rosaceana (Harris), to the main component of their pheromone blend and traces of its geometric isomer (Z11-14:Ac and E11-14:Ac, respectively) at 36 +/- 12 ng/ml air for durations of 15 and 60 min in sealed Teflon chambers with continuous air exchange significantly reduced peripheral sensory responses to these compounds as measured by electroantennograms (EAGs). The EAG responses of C. rosaceana to all tested dosages of pheromonal stimuli and blank controls were lowered by 55-58% and made a linear recovery to 70-100% of the pre-exposure amplitude within 12.5 min at a rate of 3-4 %/min. Exposures of 5 min were insufficient to maximally adapt C. rosaceana; however, exposures of 15 and 60 min reduced sensory responsiveness to the same minimum. In contrast, EAG responses of redbanded leafroller, Argyrotaenia velutinana (Walker), after identical pheromone exposure for 5 and 60 min yielded no long-lasting peripheral sensory adaptation as measured by EAGs, even though this species shares the same main pheromone components with C. rosaceana. We postulate that the long-lasting peripheral adaptation observed for C. rosaceana is a mechanism that impedes central nervous system habituation in this species. In contrast, A. velutinana may be more susceptible to central nervous system habituation because it lacks the capacity for minutes-long adaptation. We propose that long-lasting adaptation may be a mechanism explaining some of the variation in efficacy of pheromone-based mating disruption across taxa. PMID- 12737264 TI - Identification of male-specific chiral compound from the sugarcane weevil Sphenophorus levis. AB - Comparative gas chromatographic analyses of airborne volatiles produced by males and females of the sugarcane weevil Sphenophorus levis, showed one male-specific compound. Gas chromatography-mass spectrometry data indicated an aliphatic alcohol that was identified as 2-methyl-4-octanol. Both optical isomers were synthesized in five steps by employing commercially available (R)- and (S)-2.2 dimethyl-1,3-dioxolane-4-methanol as starting material. Enantiomeric resolution by gas chromatography with a chiral column demonstrated that the natural alcohol possessed the S configuration. Preliminary indoor observations suggested that the alcohol elicited aggregation behavior among adults. The same compound has been previously described as an aggregation pheromone in several other curculionid species. PMID- 12737267 TI - Does pheromone-based aggregation of codling moth larvae help procure future mates? AB - In field and laboratory bioassay experiments, we show that larvae of the codling moth, Cydia pomonella, cocoon in aggregations. This aggregation behavior of fifth instar larvae prior to pupation and arrestment of eclosed adult males by mature female pupae seems to allow mating as soon as an adult female ecloses. This synchronous timing is realized because foraging fifth-instar are attracted by cocoon-spinning larvae and prepupae, but not by pupae, and because male pupae develop faster than female pupae. Eclosed males are arrested by sex pheromone that disseminates from female pupae even before adult females eclose. Communication in C. pomonella within and among developmental stages (larva-larva and pupa-adult, respectively) may be a strategy to procure mates. If so, our data add to current knowledge that attraction of mates in insects relies on communication among adults, or pupae and adults. PMID- 12737268 TI - Dose and enantiospecific responses of white pine cone beetles, Conophthorus coniperda, to alpha-Pinene in an eastern white pine seed orchard. AB - The white pine cone beetle, Conophthorus coniperda, exhibited dose and enantiospecific responses to alpha-pinene in stands of mature eastern white pine, Pinus strobus, in a seed orchard near Murphy, North Carolina, USA. (-)-alpha Pinene significantly increased catches of cone beetles to traps baited with (+/-) trans-pityol. (+)-alpha-Pinene did not increase catches of beetles to pityol baited traps and interrupted the response of beetles to traps baited with (+/-) trans-pityol and (-)-alpha-pinene. Maximal attraction of cone beetles to pityol baited traps was obtained with lures releasing (-)-alpha-pinene at a rate of 103 mg/day at 23 degrees C. Lures releasing (-)-alpha-pinene at rates lower or higher than 103 mg/day resulted in reduced catches to traps baited with (+/-)-trans pityol. The sex ratio in all catches was heavily male biased. Attraction of the clerid predator, Thanasimus dubius, to traps baited with (+/-)-trans-pityol increased significantly with the presence of alpha-pinene, irrespective of enantiomeric composition. Maximal attraction of T. dubius to pityol-baited traps occurred with devices releasing (-)-alpha-pinene at the highest rate tested, 579 mg/d at 23 degrees C, a sub optimal rate for cone beetles. PMID- 12737269 TI - Discovery of 3-methyl-2-buten-1-yl acetate, a new alarm component in the sting apparatus of Africanized honeybees. AB - We analyzed the alarm pheromone components from five colonies of Africanized honeybees and three colonies of European honeybees collected in Mexico. Analyses revealed a novel alarm pheromone component that was only present in appreciable quantities in the Africanized bee samples. Analysis of the mass spectrum and subsequent synthesis confirmed that this compound is 3-methyl-2-buten-1-yl acetate (3M2BA), an unsaturated derivative of IPA. In Africanized honeybees, sampling from stings of guards showed that 3M2BA was present at levels of 0-38% the amount of isoamyl acetate (IPA). Behavioral assays from three colonies each of Africanized and European bees showed that 3M2BA recruited worker bees from hives of both Africanized bees and European bees at least as efficiently as isopentyl acetate IPA, a compound widely reported to have the highest activity for releasing alarm and stinging behavior in honeybees. However, a mixture of of 3M2BA and IPA (1:2) recruited bees more efficiently than either of the compounds alone. None of the compounds differed in their efficacy for inducing bees to pursue the observers. PMID- 12737271 TI - Chemical attractants in horseshoe crab, Limulus polyphemus, eggs: the potential for an artificial bait. AB - Horseshoe crabs, Limulus polyphemus, are the preferred bait in the eel and conch fisheries along the east coast of the United States. However, recent management measures have restricted the availability of horseshoe crabs to commercial fisheries, creating the need for sustainable, alternative bait sources. In this study, we examined the chemistry underlying the predator-prey attraction to determine if specific, isolable attractant metabolites from the horseshoe crab could be identified and characterized for incorporation into an artifical bait. Initial assays with the mud snail, Hyanassa obsoleta, suggested that the chemoattractants were concentrated in L. polyphemus eggs. Chemical analyses and biological assays of the egg extract indicated the primary cue was a heat-stable, proteinaceous compound (>10 kDa). A carbohydrate-rich fraction of low molecular mass (< 10 kDa) also enhanced mud snail chemotaxis. Analysis of egg digests with SDS-PAGE confirmed the presence of glycoproteins or carbohydrate-binding proteins in the horseshoe crab egg extract. Because the attractant appears to be a complex protein or glycoprotein, conventional chemical synthesis is unlikely. However, the tools of modem biotechnology offer the potential to produce this attractant in a system independent of the horseshoe crab. Such an attractant could be incorporated into an artificial bait, providing an ecologically sound alternative for commercial eel and whelk fisheries. PMID- 12737270 TI - Mating behaviour and evidence for a female released courtship pheromone in the signal crayfish Pacifastacus leniusculus. AB - The mating behaviour of the signal crayfish, Pacifastacus leniusculus (Dana), has been categorised into seven distinct stages: orientation; contact; seizure; turning; mounting; spermatophore deposition; and dismounting. These categories were used in the development of a bioassay to test whether mature females release a sex pheromone during the breeding season to which males respond. Water conditioned by 1) mature females (MF water), 2) immature females (IF water) and 3) freshwater (C water), were injected through standard aquarium air-stones into tanks containing mature males. The behaviour of the male was recorded on video for 15 minutes before and after injection of each test water. The time that the males spent exhibiting the three behavioural categories of 'quiescent,' 'motile' and 'handling' of the air-stone, were recorded. 'Handling' of the air-stone included the mating behaviours of seizure, mounting, and spermatophore deposition onto the surface of the air-stone. Males exposed to MF water exhibited significantly increased levels of motile activity and handling behaviour, than males exposed to C water or IF water. These results demonstrate for the first time the presence of a sex pheromone, released during the breeding season by mature females, that stimulates courtship and mating behaviour in male P. leniusculus. PMID- 12737272 TI - Behavioral responses of sexually active mud snails: kariomones and pheromones. AB - We postulated that kariomones and pheromones function to organize egg capsule deposition and breeding aggregations of mud snails (Ilyanassa obsoleta). Sexually active male and female mud snails were exposed to water-born odors from juvenile snails, nonaggregated female snails, oysters, egg capsule depositing snails, sexually active males, sexually active females, and copulating snails and tested for a follow response using a ring assay. Neither sex responded to juvenile odor or to odor of sexually inactive females. Snails turned and followed water born odors from oysters, when tested with the odors of sexually active males, sexually active females, and copulating pairs, we found sex-specific differences in responses. Sexually active females followed the odor of sexually active males and ignored sexually active female odor. Sexually active males followed the odor of sexually active females and ignored sexually active male odor. Both sexes followed the odor of copulating pairs and egg capsule depositing snails. Snail activities in breeding aggregations are organized by at least three pheromones and one kariomone, and reflect the multifunctionality of the aggregations. PMID- 12737273 TI - Male-specific EAD active compounds produced by female European chafer Rhizotrogus majalis (Razoumowsky). AB - Chemical investigation of a female balloon-like organ of the European chafer, Rhizotrogus majalis (Razoumowsky), with GC-EAD has resulted in the identification of female-specific compounds, (R)-3- hydroxybutan-2-one, (2R,3R)-2,3-butanediol, and meso-2,3-butanediol that are specifically EAD-active with male antennae. No behavioral role for any of the EAD active compounds could be discerned with this species. PMID- 12737275 TI - Pathology and aspects of pathogenesis in pulmonary arterial hypertension. AB - Pulmonary Arterial Hypertension (PAH) is a term which has been recently defined at the WHO-meeting in Evian and includes Primary Pulmonary Hypertension (PPH), as well as PAH related to specific pathological conditions. The poor prognosis of the disease seems to be related to a peculiar pathological anatomy, consisting of characteristic lesions of small, muscular pulmonary arteries. In addition to common hypertrophy of the tunica media, other proliferative lesions such as intimal thickening or plexiform lesions are summarized under the term of plexogenic arteriopathy. Moreover, in situ thrombosis and rarely isolated pulmonary arteritis can be observed in lungs of patients displaying PAH. Different pathomechanisms explaining these morphological changes of pulmonary vasculature have been discussed in the past, including endothelial and thrombocytic dysfunction, deregulated vasoconstriction, coagulation abnormalities, or cancer-like growth. Furthermore, latest studies on the importance of inflammatory mediators, so-called chemokines, in the lungs of PAH patients have brought a possible inflammatory component of the disease into consideration. Finally, recent identification of responsible gene mutations in subgroups of patients, as well as latest developments in genetic screening have shed some light into the modus of inheritance. Nevertheless, the role and impact of these different pathomechanisms have yet to be better defined. PMID- 12737274 TI - Chronic obstructive pulmonary disease (COPD): new insights on the events leading to pulmonary inflammation. AB - The presence of a heterogeneous infiltrate of macrophages, neutrophils and CD8+ Tc1 cells is a characteristic feature in the lung of patients with chronic obstructive pulmonary disease (COPD). This paper points out the contribution of different inflammatory cells and mediators to the pathogenesis and natural history of COPD. We will comment on data suggesting that CD8 cytotoxic T cells with an activated Tc1 phenotype migrate from the secondary lymphoid tissue to pulmonary tissue damaged by smoke or infective agents. On the basis of the knowledge of the pathophysiology of immunologic events, drugs that can potentially block the inflammation leading to the disability of COPD are being investigated. Long-term study in a large number of patients with COPD will be needed to verify the impact of a number of anti-inflammatory compounds in this increasingly common disease. PMID- 12737276 TI - Analysis of IL6 and IL1A gene polymorphisms in UK and Dutch patients with sarcoidosis. AB - BACKGROUND: Proinflammatory cytokines are a major determinant in the inflammatory events leading to sarcoidosis. Genetic variations in the genes encoding these cytokines might contribute to sarcoidosis susceptibility, disease severity and outcome. METHODS: In the present study we genotyped two clinically well-defined cohorts of Caucasian sarcoidosis patients from different European countries (each with their own controls) for the following polymorphisms using SSP-PCR: IL6 174(G/C), IL6 intron 4(A/G) and IL1A-889(C/T). In total, 516 individuals were studied (147 UK + 102 Dutch patients, 101 UK + 166 Dutch controls). Disease severity data at presentation included chest radiographic stage, FVC, DL(CO), and extrapulmonary manifestations. Disease progression was evaluated on follow-up chest radiographs and sequential lung function measurements (2, 4 years). RESULTS: No differences in genotype, carriage and allele frequencies of the investigated polymorphisms were found in either of the populations. Analysis of genotype data in relation to disease severity data, however, showed a slightly increased carrier frequency of the rarer-174C allele in patients with Stage IV sarcoidosis (p = 0.03, Pc = 0.09). Pulmonary function progression analysis did not reveal significant associations. CONCLUSIONS: Although the investigated polymorphisms are unlikely to contribute to sarcoidosis susceptibility, the IL6 174C allele might have a role in the genetics underlying sarcoidosis severity or the progression towards pulmonary fibrosis in a particular subgroup. PMID- 12737277 TI - Increased proportion of CD8+ T-lymphocytes in the paratracheal lymph nodes of smokers with mild COPD. AB - Previous studies have shown an increased number of inflammatory cells and, in particular, of CD8+ T lymphocytes, in central airways, peripheral airways, lung parenchyma and pulmonary arteries of smokers with COPD. In this study we investigated whether this inflammatory process is restricted to the lung tissue or whether a similar process is also present in the lymph nodes of these subjects. We examined paratracheal lymph nodes obtained from 6 smokers with COPD (FEV1/VC < 88% predicted and FEV1/FVC < 70% both before and after 200 microg of inhaled salbutamol) and 6 smokers without COPD (FEV1/VC > 88% predicted and FEV1/FVC > 70%) undergoing lung resection for localised pulmonary lesions. By immunohistochemistry we quantified CD4+ and CD8+ T-lymphocytes in the lymph nodes. Smokers with COPD had a decreased ratio CD4/CD8 compared to smokers without COPD. When all subjects were considered together, the ratio CD4/CD8 showed a positive correlation with the values of FEV1/VC and a negative correlation with cigarette consumption. In conclusion, smokers with COPD have an increased proportion of CD8+ cells in the lymph nodes, indicating that a T lymphocyte pattern similar to that present in the lung tissue is also present in the lymph nodes of these subjects. This finding suggests that, in COPD, the polarisation of the immune response may occur in the regional lymph nodes, possibly as a consequence of the presentation of an endogenous antigen that remains unknown. PMID- 12737278 TI - Impact of pain in a Dutch sarcoidosis patient population. AB - BACKGROUND AND AIM: Although pain is prevalent in sarcoidosis, this has never been studied systematically. The aim of the present study was to evaluate the presence and impact of pain in sarcoidosis. METHODS: Members from the Dutch Sarcoidosis Society without co-morbidity (n = 821) participated in this study. The World Health Organisation Quality of Life assessment instrument (WHOQOL-100) was completed, as well as a symptom inventory questionnaire addressing the presence of various categories of pain, i.e., muscle pain, chest pain, abdominal pain, arthralgia, and/or headache. RESULTS: Pain was reported by 594 patients (72.4%). Arthralgia was experienced most frequently (53.8%), followed by muscle pain (40.2%), headache (28.0%) and chest pain (26.9%). The number of types of pain a patient was suffering from (ranging from 0-5) was related to the WHOQOL- 100 Pain and Discomfort scale (r = 0.49, p < 0.001). Patients with more types of pain had lower quality of life (QOL). In addition, the total amount of experienced pain categories was associated with the WHOQOL-100 domain Level of Independence (r = -O.43, p < 0.001), and the facet Energy and Fatigue (r = -0.38, p < 0.001). The number of types of pain was predicted by using analgesics, psychological/neurological medication, NSAIDs, being female, indicating to feel tired, more negative feelings and less energy (F(7.635) = 35.2, p < 0.001; R2 = 27.9%). CONCLUSIONS: Pain appeared to be a major problem in sarcoidosis, especially arthralgia. Although negative feelings and fatigue were related to pain, it could not fully explain pain. Future studies are needed to address mechanisms of pain, pain behaviour, and the best therapeutic approach to pain in sarcoidosis. PMID- 12737279 TI - Anergy to tuberculin in sarcoidosis is not influenced by high prevalence of tuberculin sensitivity in the population. AB - BACKGROUND AND AIM: To study the pattern of tuberculin sensitivity and its interpretation to diagnose sarcoidosis in the presence of a high prevalence of Mantoux positivity in the general population in India. METHODS: Tuberculin sensitivity was examined in two groups of subjects: Group I included 50 newly diagnosed patients with pulmonary sarcoidosis and two control subjects with pulmonary ailments other than sarcoidosis or tuberculosis for each case; Group II included 62 new patients with sarcoidosis and 130 healthy volunteers as control subjects. Mantoux test was performed using 1 TU tuberculin. The test was repeated with 5 TU tuberculin for those cases and controls who were anergic to 1 TU among the group II subjects. Sensitivity and specificity for a "negative Mantoux test" in sarcoidosis were calculated at different cutoff points. The influence of prior BCG vaccination status on the performance of Mantoux test was also studied. MEASUREMENTS AND RESULTS: Forty-six (92%) of the 50 patients with sarcoidosis in Group I and 55 of the 62 (88.7%) in Group II did not show any reaction to Mantoux test, compared to 32 (32%) and 21 (16.2%) controls, respectively in groups I and II (P < 0.001). Thirty-nine (70.9%) of the 55 patients and 6 (28.6%) of the 21 controls from the group II subjects who were tested again with 5 TU did not show any reaction. A negative Mantoux test had a high sensitivity value at all the cut off points for the diagnosis of sarcoidosis. BCG vaccination administered during childhood had no correlation with a negative Mantoux reaction. CONCLUSIONS: Tuberculin anergy in sarcoidosis is not influenced by the rate of Mantoux positivity in the general population. A positive Mantoux test (irrespective of the size of reaction) in a suspected case of sarcoidosis should arouse strong suspicion of an alternate or an additional diagnosis of tuberculosis. PMID- 12737280 TI - Sarcoidosis in Denmark 1980-1994. A registry-based incidence study comprising 5536 patients. AB - BACKGROUND AND AIM: To evaluate the incidence of sarcoidosis in Denmark 1980 1994. METHODS: Patients with a diagnosis of sarcoidosis were identified from the Danish National Patient Registry. The file contained information about the year in which the diagnosis was reported, gender, age, and residential county. RESULTS: 5536 persons (2816 men) with sarcoidosis were registered. Median age in men was 38 years, in women 45 years. The male/female incidence ratio was 1.06. The incidence (per 100,000 person years) declined gradually from 8.1 in 1980-1984 to 6.4 in 1990-1994. The overall incidence in 1980-1994 was 7.2. Incidence rates 1980-1994 increased from eastern to western parts of Denmark: Zealand 5.7, Funen 6.8, and Jutland 8.4. The peak incidence in men occurred at 30-34 years of age (14.8). Women displayed two similar peak incidences at 25-29 years of age (10.5) and at 65-69 years of age (11.0). CONCLUSION: Incidence rates in the present study are lower compared with previous mass-screening surveys showing an incidence rate of 13.8 (in persons examined). Peak incidences occurred at higher ages in both men and women. Previous surveys showed peak incidences at 20-25 years in men and at 25-30 years and 50 years in women. The overall incidence of sarcoidosis in Denmark ranges from 7-10 per 100,000 person years. With 5,500,000 residents, the expected number of new cases of sarcoidosis in Denmark is between 385 and 550 per year. PMID- 12737281 TI - Imbalance of serum cytokine network in sarcoid patients: index of sarcoidosis relapse? AB - BACKGROUND AND AIM OF THE WORK: The follow-up of sarcoidosis is usually performed by invasive tools such as thoracic biopsy, supported by Broncho-Alveolar Lavage (BAL), Gallium-scintigraphy and Gadolinium Magnetic Resonance Imaging (Gd-MRI) to evaluate the degree of disease. Its pathogenesis can be ascribed to an accumulation of cells due to the disregulation of the immune system which involves Th1/Th2 cells as well as the soluble factors they generate. We evaluated serum cytokine and membrane marker levels (cytokine network) in sarcoid patients in order to study the correlation with the disease activity, in particular with the occurrence of sarcoidosis relapses. PATIENTS AND METHODS: Seven sarcoid patients clinically stable without therapy, at different stages, were enrolled in our study. Cytokine and membrane marker serum values were measured monthly for 24 months, after a six-month period of run-in, by ELISA assay and MoAb indirect immunofluorescence, respectively. RESULTS: In some patients at the first and second stages of the disease we observed MCP-1 and inflammatory cytokine peaks and, moreover, MCP-1 values were increased before other cytokine values. After three subsequent increases of these parameters were observed, according to our personal experience, Gadolinium-MRI confirmed the presence of an increase of the lesion and therefore our hypothesis of sarcoidosis relapse. Only one patient at stage III showed constantly elevated values of fibro-angiogenic cytokines and membrane receptors of the TNF receptor family. CONCLUSIONS: Cytokine network monitoring could be a non-invasive means of following up the clinical course of sarcoidosis. PMID- 12737282 TI - Idiopathic bronchiolitis mimicking diffuse panbronchiolitis. AB - Diffuse panbronchiolitis (DPB) is a distinct sinobronchial syndrome which has been almost exclusively reported in East Asia; only a few cases have been observed in non-Asians so far. Since 1990, when one of us described the first European case, we have screened all patients presenting with a clinical, physiopathological and radiological pattern compatible with DPB, in order to assess if the paucity of proven cases in Europe could reflect a lack of awareness of this disease among both pulmonologists and pathologists. Over a 10-years period we identified four additional DPBs, but also interestingly observed five cases of "idiopathic" bronchiolitis (IB) that had both clinical symptoms and high resolution computed tomography (HRCT) findings indistinguishable from those of DPB, but did not show the typical histological changes. All these patients were Caucasians and presented with dyspnea, productive cough, an obstructive or mixed impairment on pulmonary function tests and chest HRCT features strongly suggesting a grade II to IV DPB. We suggest that diseases classified as IB in this study might be homogeneous entities which are clinically and radiologically similar to DPB. PMID- 12737283 TI - Sarcoidosis complicated with IgA nephropathy. AB - Though IgA nephropathy (IgAN) and sarcoidosis have been reported simultaneously in patients occasionally, it remains controversial whether there is any association between the two. In this report we describe our experience with one definite and two subclinical cases of IgAN diagnosed during the clinical course of sarcoidosis. Our findings suggest a possible association between these disorders, and a higher incidence of subclinical IgAN among sarcoidosis patients than among the general population. We also discuss the common immunological background shared by these two diseases. PMID- 12737284 TI - Ambivalence of CD4 lymphocytopenia in sarcoidosis. PMID- 12737285 TI - A case of nonspecific interstitial pneumonia associated with thymoma. PMID- 12737286 TI - Severe alveolar haemorrhage in legionella pneumonia. PMID- 12737287 TI - Cervical soft tissue metastases in head and neck cancer. PMID- 12737288 TI - Psychological aspects of Meniere's disease. PMID- 12737289 TI - A head and neck cancer patient dies! Why perform an autopsy: for the relatives, for the clinicians or for the pathologists? AB - It is no secret that autopsy rates at most hospitals worldwide--both teaching and community hospitals--have declined precipitously in recent decades, but is this a desirable state of affairs? This article explores this issue from three viewpoints: that of the family members who grant permission for autopsies; that of the clinicians who seek permission for the autopsy to be carried out; and that of the pathologists who actually perform the post-mortem examination. Family concerns about (the sometimes tangential) matter of organ retention following autopsy have recently been highlighted in Europe, with an accompanying negative overall impression of the autopsy being conveyed by many outlets of the popular media. Clinicians will often concede that they feel somewhat distanced from the whole process of autopsy, and so do not hold it in such high esteem as their predecessors once did decades ago. Pathologists at present often perform autopsies as "additional duties" to be fitted in around their central functions, and so do not see them as a primary task to be accomplished. However, there are reasons why this may be detrimental to patient care, including in particular the fact that clinical/radiographic diagnoses are sometimes not confirmed by the results of a complete autopsy. Suggestions for improving the autopsy rate--in particular amongst head and neck cancer patients--are discussed, and include performance of a more rapid limited autopsy and the designation of specialist pathologists in head and neck cancer to carry out autopsies of these patients as an extension of their clinical duties. One conclusion seems inescapable: to increase autopsy rates, the status of the procedure will necessarily have to be upgraded from that of "afterthought/perfunctory task" to that of "consultation", with all of the shifts in attitude such a modification would entail. PMID- 12737290 TI - Photochemically induced double lateral wall lesions in the guinea pig cochlea. AB - OBJECTIVE: Multiple patches of atrophy have been reported in the stria vascularis (SV) in elderly persons with presbycusis The aim of this study was to investigate the correlation between sensorineural hearing loss and this strial condition. MATERIALS AND METHODS: We established a new animal model comprising two small lesions in the SV in the second turn of the cochlea by means of photochemical reaction. Using this model, we investigated morphological and physiological changes in the cochlea at 3, 7 and 14 days after SV damage. RESULTS: Scanning electron microscopy studies revealed that the strial cells between the two damaged areas of the SV remained intact, although the outer hair cells (OHCs) facing the intact SV area were damaged. Furthermore, damage to the first and second rows of OHCs gradually progressed throughout the 14-day observation period. The endocochlear potential (EP) measured at a point midway between the 2 lesions at 3 and 7 days was found to be significantly lower compared with control values, but had returned to a normal level at 14 days CONCLUSION: The reversible EP change and localized OHC loss seen in the present investigation may help to understand acute idiopathic or progressive sensorineural hearing loss. PMID- 12737291 TI - Effect of uridine 5'-triphosphate on mucin and lysozyme expression in human middle ear epithelial cells. AB - OBJECTIVE: Extracellular uridine 5'-triphosphate (UTP) regulates a variety of biological functions in the airway epithelium, including chloride and fluid transport, mucociliary clearance and mucin secretion via the P2Y purinergic receptors. This study was undertaken to investigate which P2Y purinergic receptors are expressed in normal human middle ear epithelial (NHMEE) cells. We also determined the levels of mucin and lysozyme secretion and their mRNA expressions following stimulation with UTP in passage-2 cultured NHMEE cells. MATERIAL AND METHODS: An immunoblotting assay was performed for quantitation of mucin and lysozyme proteins and RT-PCR for their gene levels after treatment with UTP was done in normal human middle ear epithelial cells. RESULTS: Middle ear epithelial cells expressed P2Y1, P2Y2, P2Y6, P2Y11, and P2Y12 receptors but not P2Y4 receptor. Apically applied UTP induced increased mucin and lysozyme secretion, as measured by dot blotting. In contrast, UTP did not enhance mucin and lysozyme mRNA expression until 72 h after treatment. CONCLUSION: This study suggests that UTP acts as a secretogogue on mucin and lysozyme secretion in NHMEE cells via the P2Y2 and/or P2Y6 receptor. PMID- 12737292 TI - Detection of human papillomavirus in temporal bone inverted papilloma by polymerase chain reaction. AB - There is debate about the role of human papillomavirus in the induction of rare inverted papillomas involving the temporal bone and in the higher recurrence rates and association with squamous cell carcinoma of temporal bone inverted papillomas compared with sinonasal inverted papillomas. An exhaustive review of the literature revealed that eight cases of temporal bone inverted papilloma have been analysed for human papillomavirus. None of the cases studied with in situ hybridization proved positive. Only one case was found to be positive using the more sensitive polymerase chain reaction assay. We present the first two cases of Schneiderian-type papilloma involving the temporal bone to be analysed by type specific polymerase chain reaction methods for human papillomavirus. PMID- 12737293 TI - Chromosome 8 aneuploidy in acquired cholesteatoma. AB - OBJECTIVE: To investigate the incidence of chromosome 8 aneuploidy in acquired cholesteatoma. MATERIAL AND METHOD: Cholesteatoma tissue and postauricular skin as a control were surgically obtained from 12 patients with acquired cholesteatoma. Fluorescence in situ hybridization (FISH) analysis using a chromosome 8-specific alpha-satellite DNA probe was performed on the interphase nuclei. Two hundred cells were analyzed for each of the samples. RESULTS: Chromosome 8 aneuploidy was found in 9/12 patients whereas a normal cell structure with 2 signals was observed in the remaining 3 patients. In samples with chromosome 8 aneuploidy, the mean proportion of aneuploidy was 25.6%, including monosomy (3.2%), trisomy (16.1%), tetrasomy (4.9%) and more than tetrasomy (1.4%). The number of aneuploid cells in recurrent cases was more than that in non-recurrent cases. CONCLUSION: A numerical aberration of chromosome 8 was found in patients with acquired cholesteatoma. Our results support the hypothesis that chromosome 8 may be a prognostic factor for cholesteatoma and an indicator in the follow-up of patients with cholesteatoma. PMID- 12737294 TI - Cholesteatoma epithelium is characterized by increased expression of Ki-67, p53 and p21, with minimal apoptosis. AB - OBJECTIVE: To investigate differences in cell proliferation, cell cycle arrest and apoptosis between cholesteatoma and control skin. MATERIAL AND METHODS: Immunohistochemical sections of 15 cholesteatoma and 15 paired control retro auricular skin samples were examined for Ki-67, p53, p21 and active caspase 3, using image analysis, as well as for DNA fragmentation. RESULTS: The retro auricular skin samples contained 5.7% +/- 3.6%, Ki-67-positive cells and showed a normal expression pattern. In the cholesteatoma epithelium 11.7% +/- 9.5% of the cells were Ki-67-positive and these cells were dominantly expressed in the basal and parabasal cell layers. Retro-auricular skin contained 5.8% +/- 5.4% p53 positive cells and 1.0% +/- 0.9%, p21-positive cells. In the cholesteatoma epithelium 17.8% +/- 12.3% of the cells were p53-positive and 14.3% +/- 11.6% were p21-positive The expression of Ki-67, p53 and p21 differed significantly between the two groups (p < 0.05). In the cholesteatoma epithelium a positive correlation was found between p53 and p21 expression (p = 0.016). Active caspase 3 positivity and DNA fragmentation were rarely seen in the cholesteatoma epithelium. CONCLUSION: Our results indicate that increased cell proliferation in cholesteatoma epithelium is accompanied by an increase in p53 and p21 protein levels, whilst apoptosis is minimal. PMID- 12737295 TI - IL-5 and eotaxin levels in middle ear effusion and blood from asthmatics with otitis media with effusion. AB - OBJECTIVE: The purpose of this study was to evaluate eosinophil infiltration as well as IL-5 and eotaxin levels in middle ear effusion (MEE) and blood from otitis media with effusion (OME) patients with asthma and to compare the findings with those from OME patients without asthma (control group). MATERIAL AND METHODS: Levels of IL-5 and eotaxin in MEE and blood were measured by means of enzyme-linked immunosorbent assay. RESULTS: 5 levels in MEE were significantly higher than those in blood in both groups of patients and in OME patients with asthma than in the control group. In addition, in OME patients with asthma, there was a significant correlation between the percentage of eosinophils and IL-5 levels in MEE. Eotaxin levels in blood were significantly higher than those in MEE in both groups of patients and in OME patients with asthma than in the control group. In addition, in OME patients with asthma, the percentage of eosinophils and eotaxin levels in blood tended to correlate, but did not reach statistical significance. CONCLUSION: These data suggest that, in OME patients with asthma, eosinophilia in MEE depends more on IL-5 than on eotaxin, and that eotaxin may play an important role in the mobilization of eosinophils from the bone marrow into the blood. PMID- 12737296 TI - Prediction of progression from atypical to definite Meniere's disease using electrocochleography and glycerol and furosemide tests. AB - OBJECTIVE: To investigate whether electrocochleography (ECochG) and glycerol and furosemide tests could predict progression from atypical to definite Meniere's disease (MD). MATERIAL AND METHODS: ECochG and glycerol and furosemide tests were performed in 1569 patients with various cochleovestibular diseases, including definite MD, atypical MD, syphilitic labyrinthitis, delayed endolymphatic hydrops, sudden hearing loss, cochleovestibulopathy and sensorineural hearing loss. Patients with atypical MD were divided into five categories based on their symptoms. RESULTS: A total of 115/118 patients (97%) with definite MD who underwent all 3 tests showed a positive result in at least 1 test. Ninety-nine patients who did not satisfy the diagnostic criteria of definite MD but had vertigo and/or hearing loss at the first visit subsequently progressed to definite MD. It was retrospectively found that 92% of patients showed at least 1 positive finding in these 3 tests at the initial stage. In those patients who showed a negative test result in either ECochG or the glycerol test, the possibility of progression to definite MD was low. CONCLUSION: The combination of ECochG and the glycerol and furosemide tests was helpful in diagnosing endolymphatic hydrops (ELH). ECochG and the glycerol test were effective tools for predicting the progression to definite MD in patients with atypical MD, sudden hearing loss and other cochleovestibular diseases. Our test results also indicated that the pathological state of atypical MD included both non-ELH and ELH. PMID- 12737297 TI - Clinical characteristics of audio-vestibular impairment in Meniere's disease: does vestibular function deteriorate in accordance with cochlear function? AB - The clinical course of Meniere's disease and the outcomes of functional examinations were found to be correlated, and a difference was found between the progressions of vestibular and cochlear dysfunction. Neither the total number of definitive vertiginous spells nor the duration of illness correlated significantly with the degree of audio-vestibular dysfunction. In contrast, frequent vertiginous spells were clearly associated with preservation of audio vestibular function. Moreover, the clinical characteristics of those patients who experienced less than one definitive vertiginous spell per year were distinct from those of other patients, although both groups showed the typical symptoms of Meniere's disease. PMID- 12737298 TI - Effect of histamine on the permeability of the nasal mucosa in vivo. AB - OBJECTIVE: Histamine is one of the chemical mediators released during the acute phase of allergic rhinitis and is considered to cause the increase in epithelial permeability observed. We tried to examine the effect of histamine on nasal mucosal permeability in vivo. MATERIAL AND METHODS: Histamine at different concentrations was administered to the nostrils of healthy subjects and the nasal transepithelial potential difference (PD) was measured. We also examined nasal mucosal permeability by means of a histochemical technique using horseradish peroxidase (HRP) in guinea pigs. RESULTS: Administration of 10(-1) M histamine significantly reduced the nasal PD in healthy subjects. After administration of 5.4 x 10(-1) M histamine to the noses of guinea pigs, most ofthe intercellular spaces showed positive reactions to HRP and this effect was significantly inhibited by pretreatment with mepyramine and the antihistamine bepotastine besilate. CONCLUSION: These results indicate that histamine plays an important role in the change in mucosal permeability observed in allergic rhinitis in vivo via the histamine H1 receptor. PMID- 12737299 TI - Hyaluronan and alpha-atrial natriuretic polypeptide in human nasal polyps: contributing factors to oedema formation and polyp growth? AB - OBJECTIVES: To identify and localize hyaluronan (HYA) and alpha-atrial natriuretic polypeptide (ANP) in human nasal polyps and to measure the HYA concentrations. MATERIAL AND METHODS: Twelve nasal polyps were collected during routine polypectomies and processed histochemically and biochemically to determine the occurrence of HYA. The distribution of ANP was investigated using an immunocytochemical method. RESULTS: HYA was unevenly distributed, being found abundantly in the surface epithelium and basement membrane and around fibres and vessels in the lamina propria. It was also present around seromucinous glands and in the secretion of cysts in the stroma. The HYA concentration was 1,000-fold higher than in serum. ANP was abundant in the apical part of ciliated surface epithelial cells and extracellularly in the basement membrane. In the stroma, ANP was confined to apical acinar cells of the seromucinous glands. CONCLUSIONS: Osmotically active HYA and numerous ANP-immunoreactive cells, active in fluid and/or ion transport functions, are present in human nasal polyps. These substances may well be involved in oedema formation and the successive growth of nasal polyps. The high concentrations of HYA in nasal polyps may be of clinical significance for the future development of a local enzyme treatment for nasal polyposis. PMID- 12737300 TI - Histopathologic assessment of fungal involvement of the paranasal sinuses in Turkey. AB - OBJECTIVE: To assess paranasal sinus material histopathologically for the presence of fungus. MATERIAL AND METHODS: Paraffin-embedded archival biopsy samples of patients who underwent endonasal sinus surgery between 1992 and 2002 were retrospectively assessed for the presence of fungi. Hematoxylin-eosin stained sections of the materials were re-evaluated, and Gomori's methanamine silver stain was also applied as required. RESULTS: Fungus (Aspergillus) was detected histopathologically in only 21476 patients, both of whom were immunocompetent. One patient was considered to have chronic indolent sinusitis and the other allergic fungal sinusitis. CONCLUSIONS: Although histopathologic assessment is one of the most important diagnostic tools, on its own it may lead to underestimation of fungal involvement of the paranasal sinuses. Alternatively, fungal involvement of the paranasal sinuses may be very infrequent in Turkey. PMID- 12737301 TI - Steroid injection for Reinke's edema using fiberoptic laryngeal surgery. AB - OBJECTIVE: Since 1990, we have performed steroid injections into the vocal fold under topical anesthesia using fiberoptic laryngeal surgery (FLS) in an outpatient clinic. The aim of this study was to retrospectively assess the usefulness of this treatment method in 44 patients with mild Reinke's edema. MATERIAL AND METHODS: Using fiberoptic monitoring of the larynx, a curved injection needle was inserted via the oral cavity and triamcinolone acetonide was injected into Reinke's space of the bilateral vocal fold. RESULTS: Remission or improvement was observed in almost all patients in terms of both patients' self rating of hoarseness and endoscopic vocal fold findings The maximum phonation time was a mean of 9.0 s before operation and 11.4 safter operation, and this increase was significant (p < 0.01). Voice pitch also improved, from 168 to 181 Hz, in female patients, and this increase was also significant (p < 0.05). CONCLUSION: Steroid injection is considered to be useful for treating mild Reinke's edema. PMID- 12737302 TI - Stomal recurrence: pathophysiology, treatment and prevention. PMID- 12737303 TI - Esthesioneuroblastoma: a Danish demographic study of 40 patients registered between 1978 and 2000. AB - OBJECTIVE: A retrospective review of all diagnosed cases of esthesioneuroblastoma registered in Denmark between 1978 and 2000 was carried out in order to obtain epidemiological data and optimize national treatment guidelines. MATERIAL AND METHODS: Forty cases were verified histologically and included in the analysis Epidemiological and histopathological data were evaluated in relation to the clinical outcome. RESULTS: The 40 cases represent an incidence rate of 0.4 cases/million inhabitants per year. Eight (20%) patients were classified as Kadish stage A, 13 (32.5%) as stage B and 19 (47.5%) as stage C. The histopathological findings were classified according to the grading system of Hyams The median follow-up time was 2.3 years (range 0.3-11.1 years). The 5-year crude survival rate was 61%, with a median survival of 3.1 years (range 0.3-19.2 years). The 5-year disease-free survival rate was 50%, with a median survival of 1.7 years (range 0-19.2 years). Only 3 (7%) patients had positive cervical lymph nodes at presentation. A nationwide consensus regarding treatment was seen in patients classified as Kadish stages A and B. The longest duration before the first recurrence of esthesioneuroblastoma was 5(1/2) years. CONCLUSIONS: The following therapeutic guidelines are suggested: Kadish stage A patients, surgical tumour resection and radiotherapy; Kadish stage B, surgical tumour resection and radiotherapy; Kadish stage C, surgical tumour resection via a craniofacial resection and radiotherapy combined with chemotherapy. Long-term follow-up of esthesioneuroblastoma patients is mandatory. PMID- 12737305 TI - Molecular farming of recombinant antibodies in plants. AB - Antibodies represent a large proportion of therapeutic drugs currently in development. In most cases, they are produced in mammalian cell lines or transgenic animals because these have been shown to fold and assemble the proteins correctly and generate authentic glycosylation patterns. However, such expression systems are expensive, difficult to scale up and there are safety concerns due to potential contamination with pathogenic organisms or oncogenic DNA sequences. Plants represent an inexpensive, efficient and safe alternative for the production of recombinant antibodies. Research over the last 10 years has shown that plants can produce a variety of functional antibodies and there is now intense interest in scaling up production to commercial levels. In this review, we discuss the advantages of plants over traditional expression systems, describe how antibody expression in plants is achieved and optimized and then consider the practical issues concerning large-scale molecular farming in plants. The first plant-produced therapeutic antibodies are already in clinical trials, and, given the economic benefits of this production system, we are likely to see many more recombinant antibodies produced in this manner in the future. PMID- 12737304 TI - Angiosarcoma of the submandibular salivary gland: case report and review of the literature. AB - OBJECTIVES: The purpose of this report is to describe an example of angiosarcoma (AS) of the submandibular salivary gland. The clinical and immunohistopathologic features of these lesions are also reviewed. MATERIAL AND METHODS: A 17-year-old male high school student was admitted to our clinic with a 1-year history of a slow-growing, tender mass in the left submandibular region. Physical examination on initial presentation revealed a diffuse soft tissue mass 6 cm in diameter involving the left submandibular region. MRI analysis revealed a mass located superolateral to the submandibular salivary gland, measuring 4.0 x 2.0 cm2. The mass was excised completely together with the left submandibular salivary gland. Histopathologic analysis led to a diagnosis of AS. Immunohistochemical studies were also used to determine endothelial cell differentiation. Owing to the lateralized nature of the lesion, a left functional neck dissection was performed and postoperative radiotherapy was planned. RESULTS: AS is a malignant tumor of endothelial cell origin that may occur in any region of the body. The commonest sites include the extremities and the retroperitoneal space, with only 4% of AS tumors arising in the head and neck area. The submandibular salivary gland is an extremely rare location for this tumor. Based on a literature review, this case report represents only the second reported case of AS of the submandibular salivary gland. CONCLUSIONS: In most cases, radical surgery and postoperative radiotherapy are advocated to treat patients with AS tumors, with lymph node clearance recommended in cases of lateralized lesions In some patients, distant metastasis may occur after many years, which necessitates long-term follow-up. The prognosis is poor in most cases of AS. PMID- 12737306 TI - Structure, function and evolution of haspin and haspin-related proteins, a distinctive group of eukaryotic protein kinases. AB - The haspins constitute a newly defined protein family containing a distinctive C terminal eukaryotic protein kinase domain and divergent N termini. Haspin homologues are found in animals, plants and fungi, suggesting an origin early in eukaryotic evolution. Most species have a single haspin homologue. However, Saccharomyces cerevisiae has two such genes, while Caenorhabditis elegans has at least three haspin homologues and approximately 16 haspin-related genes. Mammalian haspin genes have features of retrogenes and are strongly expressed in male germ cells and at lower levels in some somatic tissues. They encode nuclear proteins with serine/threonine kinase activity. Murine haspin is reported to inhibit cell cycle progression in cell lines. One of the S. cerevisiae homologues, ALK1, is a member of the CLB2 gene cluster that peaks in expression at M phase and thus may function in mitosis. Therefore, the haspins are an intriguing group of kinases likely to have important roles during or following both meiosis and mitosis. PMID- 12737307 TI - Role of plasminogen activator-plasmin system in tumor angiogenesis. AB - New blood formation or angiogenesis has become a key target in therapeutic strategies aimed at inhibiting tumor growth and other diseases associated with neovascularization. Angiogenesis is associated with important extracellular remodeling involving different proteolytic systems among which the plasminogen system plays an essential role. It belongs to the large serine proteinase family and can act directly or indirectly by activating matrix metalloproteinases or by liberating growth factors and cytokines sequestered within the extracellular matrix. Migration of endothelial cells is associated with significant upregulation of proteolysis and, conversely, immunoneutralization or chemical inhibition of the system reduces angiogenesis in vitro. On the other hand, genetically altered mice developed normally without overt vascular anomalies indicating the possibility of compensation by other proteases in vivo. Nevertheless, they have in some experimental settings revealed unanticipated roles for previously characterized proteinases or their inhibitors. In this review, the complex mechanisms of action of the serine proteases in pathological angiogenesis are summarized alongside possible therapeutic applications. PMID- 12737308 TI - ATP synthases: structure, function and evolution of unique energy converters. AB - A-, F- and V-adenosine 5'-triphosphatases (ATPases) consist of a mosaic of globular structural units which serve as functional units. These ion translocating ATPases are thought to use a common mechanism to couple energy of ATP hydrolysis to ion transport and thus create an electrochemical ion gradient across the membrane. In vitro, all of these large protein complexes are able to use an ion gradient and the associated membrane potential to synthesize ATP. A-/F /V-type ATPases are composed of two distinct segments: a catalytic sector, A1/F1/V1, whose three-dimensional structural relationship will be reviewed, and the membrane-embedded sector, Ao/Fo/Vo, which functions in ion conduction. Recent studies on the molecular biology of the Ao/Fo/Vo domains revealed surprising findings about duplicated and triplicated versions of the proteolipid subunit and shed new light on the evolution of these ion pumps. PMID- 12737309 TI - Murine models of colorectal cancer: studying the role of oncogenic K-ras. AB - Cancers of the intestine are amongst the most frequent tumors in the Western countries. They arise through the stepwise, progressive disruption of cellular signalling cascades which control cell proliferation, survival and differentiation. The proto-oncogene K-ras functions as an important molecular switch linking several of these signalling pathways. Activating mutations of K ras are found in about 50% of colorectal cancers, but their contribution to tumor initiation and progression is still poorly understood. Murine models provide excellent opportunities to identify and define the roles of genes involved in cancer formation and growth in the digestive tract. In this review, I will discuss the biological properties of oncogenic K-ras, its influence on cell signalling and its role in colorectal tumorigenesis based on recently established murine models. PMID- 12737310 TI - Dyskeratosis congenita. AB - Dyskeratosis congenita is an inherited skin and bone marrow failure syndrome. There are X-linked, autosomal dominant and autosomal recessive forms of the disease. The X-linked form is due to mutations in the DKC1 gene at Xq28. The encoded protein, dyskerin, is a component of both small nucleolar ribonuclear protein particles and the telomerase complex. Mutations in DKC1 mainly lead to amino acid substitutions. The autosomal dominant form of the disease is due to mutations in hTR, the RNA component of telomerase, making it likely that the disease is due to defective telomerase activity. Mutations in hTR are predicted to either disrupt secondary structure or alter the template region. The gene or genes involved in the recessive forms of the disease remain elusive, though genes whose products are required for telomere maintenance are strong candidates. PMID- 12737311 TI - The role of interleukin-17 in inducible nitric oxide synthase-mediated nitric oxide production in endothelial cells. AB - The effect of interleukin (IL)-17 on the activation of inducible nitric oxide (NO) synthase (iNOS) and subsequent production of NO was investigated. IL-17 induced NO production in both mouse and rat endothelial cells in a dose- and time dependent manner. This was paralleled by the induction of mRNA for iNOS, which was markedly down-regulated by specific antagonists of protein tyrosine kinase, p38 MAP kinase or iNOS transcription factor NF-kappaB. The expression of iNOS transcription factor IRF-1 was also induced by IL-17 and blocked by all three inhibitors, suggesting that the induction of iNOS by IL-17 might be partly exerted through IRF-1 activation. Neutralization with the specific antibody showed that endogenous IL-17 is involved in T cell-mediated NO production in endothelial cells and NO-dependent suppression of T cell growth. These data indicate that IL-17-triggered iNOS activation in endothelial cells might participate in regulation of the T cell-dependent inflammatory response. PMID- 12737312 TI - RU49953: a non-hormonal steroid derivative that potently inhibits P-glycoprotein and reverts cellular multidrug resistance. AB - Progesterone and the antiprogestin RU38486 have been reported as non-transported modulators of P-glycoprotein-mediated drug efflux. However, their hormonal properties limit their potential for clinical trials. The present work shows that some derivatives from either progesterone/RU38486 or estradiol, displaying differential interaction with hormone receptors, bind to P-glycoprotein and chemosensitize the growth of MDR1-transfected cells to vinblastine more strongly than does RU38486. Structure comparison of the compounds indicates that the highly hydrophobic estradiol derivative RU49953, which does not interact with any hormone receptor, inhibits P-glycoprotein-mediated drug efflux very efficiently, as monitored by flow cytometry, and prevents drug site photoaffinity labeling by azidopine. It induces a much higher chemosensitization than the well-known P glycoprotein modulator verapamil, which is itself more efficient than RU38486. RU49953 therefore constitutes a promising new lead for steroid-type modulators of multidrug resistance. PMID- 12737313 TI - Phylogeny, processing and expression of the rat cathelicidin rCRAMP: a model for innate antimicrobial peptides. AB - A database search identified a rat cDNA clone which phylogenetic analysis revealed to encode a cathelicidin most similar to mouse cathelicidin CRAMP. The analysis also showed that the evolutionary pattern of the cathelicidin family is lineage specific. The rat cathelicidin is called rCRAMP. Its peptide was isolated from granulocytes, and determined to be 43 amino acids long by mass spectrometry and N-terminal sequencing. Synthetic rCRAMP had antimicrobial activity. The expression of rCRAMP was investigated by reverse-transcriptase polymerase chain reaction followed by Southern hybridization and by Western blot analysis. rCRAMP was identified in granulocytes, thymus, testis, lung, mouth mucosa, tongue, oesophagus, colon, caecum and small intestine, a distribution similar to cathelicidins of mouse and human. The rat is a small laboratory animal with additional disease models available compared to the mouse. Our results open up the possibility to use the rat as a model system to study responses connected to cathelicidin expression in health and disease. PMID- 12737314 TI - Cl- affects the function of the GABA cotransporter rGAT1 but preserves the mutal relationship between transient and transport currents. AB - The effects of reducing external Cl- on the electrophysiological properties of the Na+/Cl(-)-dependent GABA transporter rGAT1 expressed in Xenopus oocytes were investigated. In agreement with a recently proposed kinetic scheme, the effects of Cl- are complex but preserve the mutual relationship that links the transport associated current, I(tr), measured in saturating GABA concentration, and the transient current, I(pre), recorded in the absence of GABA following a voltage step from the holding potential Vh to V. In particular, I(tr) (V) - I(tr) (Vh) = r integral I(pre) (V) dt, where r is the relaxation rate of I(pre) at the same membrane potential and Cl- concentration. The model also predicts a relationship between charge relaxation rate and apparent affinity for GABA, which is also verified in the presence of lowered Na+ or Cl- concentrations. In these conditions, the binding rate of GABA to the transporter is increased. All these effects are consistent with the hypothesis that interaction of the organic substrate with rGAT1 induces a conversion from a capacitive to a conductive mode of operation without strongly altering either the amount or the rate of charge movement. PMID- 12737315 TI - U7 snRNAs induce correction of mutated dystrophin pre-mRNA by exon skipping. AB - Most cases of Duchenne muscular dystrophy are caused by dystrophin gene mutations that disrupt the mRNA reading frame. Artificial exclusion (skipping) of a single exon would often restore the reading frame, giving rise to a shorter, but still functional dystrophin protein. Here, we analyzed the ability of antisense U7 small nuclear (sn)RNA derivatives to alter dystrophin pre-mRNA splicing. As a proof of principle, we first targeted the splice sites flanking exon 23 of dystrophin pre-mRNA in the wild-type muscle cell line C2C12 and showed precise exon 23 skipping. The same strategy was then successfully adapted to dystrophic immortalized mdx muscle cells where exon-23-skipped dystrophin mRNA rescued dystrophin protein synthesis. Moreover, we observed a stimulation of antisense U7 snRNA expression by the murine muscle creatine kinase enhancer. These results demonstrate that alteration of dystrophin pre-mRNA splicing could correct dystrophin gene mutations by expression of specific U7 snRNA constructs. PMID- 12737316 TI - Selective modulation of ER-beta by estradiol and xenoestrogens in human breast cancer cell lines. AB - In the last decades, substances with estrogenic activity have been dispersed into the environment. Xenoestrogens act by binding to estrogen receptors, ligand regulated transcription factors, for which two subtypes have been described, ER alpha and ER-beta, which are often coexpressed at variable amounts in different tissues. We investigated variations in the expression of ER-alpha and ER-beta mRNAs following treatment with four xenoestrogens (bisphenol A, 4-tert octylphenol, 2-hydroxybiphenyl, 4-hydroxybiphenyl) and with 17beta-estradiol in estrogen-sensitive (T47D) and estrogen-insensitive (BT20) breast cancer cell lines. Although to a variable extent, both estradiol and the tested xenoestrogens increased the expression of ER-beta mRNA, whereas a slight effect on ER-alpha was observed only in T47D cells. Upregulation of ER-beta expression by estradiol and xenoestrogens was observed only in the presence of detectable ER-alpha protein levels. These findings indicate a regulatory role for ER-beta in ER-alpha mediated transcription and a role for ER-beta in mediating xenoestrogen toxicity. PMID- 12737317 TI - Ceramide mediates growth inhibition of the Plasmodium falciparum parasite. AB - In mammalian cells, ceramide mediates death by chemotherapeutic drugs. We analysed, for the first time, the role of ceramide in inhibiting growth of the malaria-causing parasite Plasmodium falciparum. Added exogenously, ceramide significantly decreased the number of parasites, and this effect was abolished by sphingosine-1-phosphate, a biological antagonist of ceramide action. Ceramide can induce death of cancer cells by decreasing glutathione levels, and in our work it induced dose- and time-dependent depletion of glutathione in P. falciparum parasites. N-acetylcysteine, a precursor of glutathione, abrogated the cytotoxic effect of ceramide. Thus, ceramide can mediate growth inhibition of P. falciparum parasites by decreasing glutathione levels. The antimalarial drugs artemisinin and mefloquine induced the death of P. falciparum parasites by sphingomyelinase generated ceramide and by decreasing parasite glutathione levels. Altogether, ceramide was identified as a signalling molecule capable of inducing growth inhibition of P. falciparum malarial parasites. PMID- 12737318 TI - Recombinant anti-P protein autoantibodies isolated from a human autoimmune library: reactivity, specificity and epitope recognition. AB - The ribosomal P proteins are specific and important autoantigens in patients affected by systemic lupus erythematosus. In this study, we describe for the first time the selection and characterization of recombinant human monoclonal anti-P protein (auto)-antibody fragments from an autoimmune patient-derived phage display antibody library. The selected recombinant anti-P antibodies specifically recognize the P proteins in immunofluorescence assays on HEp-2 cells and in immunoblotting assays, and they immunoprecipitate the P proteins under native conditions. Using both anti-P-positive patient sera and the selected recombinant anti-P antibodies, the immunodominant epitope was determined and shown to be located at the C-terminal end of the P proteins (amino acids 111-115). Inhibition of in vitro protein translation demonstrated that interaction of the monoclonal patient-derived anti-P antibodies with their native epitope functionally inhibits the activity of the P proteins on the ribosome, confirming the notion that patient autoantibodies are often directed to the functional centre of their autoantigenic target. PMID- 12737319 TI - Ascaris nematodes from pig and human make three antibacterial peptides: isolation of cecropin P1 and two ASABF peptides. AB - Organisms co-habiting with bacteria have developed efficient bactericidal agents to control their microbe-rich environment. The Ascaris nematode lives in its final development stages in the gut of its host and is believed to feed on bacteria. Ascaris suum survive in pig intestine while A. lumbricoides is the principal species in humans. Here we show that A. suum and A. lumbricoides both produce linear (cecropin P1) and cysteine-rich (ASABF) peptides with activity against either gram-negative or gram-positive bacteria, respectively. Thus nematodes rely in part on a peptide-based antibacterial system for digestion of bacteria, which may also confer protection against infection. Cecropin P1 was previously isolated from pig intestine but we can now conclude that was due to contaminating nematodes. PMID- 12737320 TI - Dual effect of cannabinoid CB1 receptor stimulation on a vanilloid VR1 receptor mediated response. AB - Cannabinoid CB1 receptors and vanilloid VR1 receptors are co-localized to some extent in sensory neurons of the spinal cord and dorsal root ganglia. In this study, we over-expressed both receptor types in human embryonic kidney (HEK)-293 cells and investigated the effect of the CB1 agonist HU-210 on the VR1-mediated increase in intracellular Ca2+ ([Ca2+]i), a well-known response of the prototypical VR1 agonist capsaicin. After a 5-min pre-treatment, HU-210 (0.1 microM) significantly enhanced the effect of several concentrations of capsaicin on [Ca2+]i in HEK-293 cells over-expressing both rat CB1 and human VR1 (CB1-VR1 HEK cells), but not in cells over-expressing only human VR1 (VR1-HEK cells). This effect was blocked by the CB1 receptor antagonist SR141716A (0.5 microM), and by phosphoinositide-3-kinase and phospholipase C inhibitors. The endogenous agonist of CB1 and VR1 receptors, anandamide, was more efficacious in inducing a VR1 mediated stimulation of [Ca2+]i in CB1-VR1-HEK cells than in VR1-HEK cells, and part of its effect on the former cells was blocked by SR141716A (0.5 microM). Pre treatment of CB1-VR1-HEK cells with forskolin, an adenylate cyclase activator, enhanced the capsaicin effect on [Ca2+]i. HU-210, which in the same cells inhibits forskolin-induced enhancement of cAMP levels, blocked the stimulatory effect of forskolin on capsaicin. Our data suggest that in cells co-expressing both CB1 and VR1 receptors, pre-treatment with CB1 agonists inhibits or stimulates VR1 gating by capsaicin depending on whether or not cAMP-mediated signalling has been concomitantly activated. PMID- 12737321 TI - Design and evaluation of a diabody to improve protection against a potent scorpion neurotoxin. AB - Diabodies are recombinant, dimeric, antibody-based molecules composed of two non covalently associated single-chain antibody fragments that bind to an antigen in a divalent manner. In an attempt to develop more effective therapeutic molecules against scorpion venoms, we designed a diabody derived from monoclonal antibody 9C2, which neutralizes the toxicity of scorpion neurotoxin AahI in mammals. The recombinant diabody produced in the periplasm of Escherichia coli was purified to homogeneity in a single step by protein L-agarose affinity chromatography. It was functional, and possessed a high binding affinity to AahI (8 x 10(-11) M). The bivalence of the diabody was confirmed by size-exclusion chromatography, isoelectrofocussing and electron microscopic observations. Finally, the diabody showed high thermal stability in serum and demonstrated protective activity when injected intraperitoneally in mice experimentally envenomed with toxin AahI. In conclusion, the diabody format gives the 9C2 molecule advantageous properties that are particularly important for potential clinical applications in the treatment of envenomations. PMID- 12737322 TI - CD4+ T cell-mediated immunity against prion proteins. AB - The prion protein (PrP(C)) is essential for susceptibility to transmissible spongiform encephalopathies. A specific conformer of this protein (PrP(Sc)) is, according to the 'protein only' hypothesis, the principal or only component of the infectious agent, designated prion. Transmission of prions between species is often inefficient, resulting in low attack rates and/or prolonged incubation times and is ascribed to a 'species barrier' caused by differences in the amino acid sequence of PrP between recipient and donor. In this report, we demonstrate that these differences in amino acid sequence result in presentation of distinct peptides on major histocompatibility complex class II molecules. These peptides result in activation of specific CD4+ T cells which leads to the induction of an effective immune response against foreign PrP as demonstrated by antibody production. Therefore, CD4+ T cells represent a crucial component of the immune system to distinguish between foreign and self PrP. PMID- 12737323 TI - Imaging in rheumatoid arthritis--why MRI and ultrasonography can no longer be ignored. AB - Implementing the modern treatment strategy in rheumatoid arthritis (RA), i.e. early initiation and optimal adjustments of aggressive therapies, requires methods for early diagnosis and sensitive monitoring of the disease process. In rheumatoid arthritis clinical trials and routine management, conventional radiography is the pivotal method for diagnosing and monitoring structural joint damage. However, it is insensitive to bone damage at its earliest stages and totally incapable of capturing the primary feature of rheumatoid disease, the synovitis. In comparison with radiography, magnetic resonance imaging (MRI) offers assessment of bone damage with improved sensitivities to early pathology and to change. In addition, detailed assessment of soft tissue changes, including synovitis and tenosynovitis, is possible and MRI findings are of prognostic value for the long-term radiological outcome. Ultrasonography (US) is less validated than MRI, but available data suggests that US offers comparable information on both inflammatory and destructive changes in RA finger and toe joints. Issues of reliability, standardization and documentation limit its value in clinical trials, This article reviews current knowledge on conventional radiography, computed tomography, MRI and US for assessment of peripheral joints in RA. The rationale is provided for MRI being the new gold standard for assessment of RA joints and US becoming a routine bedside tool for improved joint assessments and injections by rheumatologists. Pursuing the goal of improving patient care and disease outcome, rheumatologists can no longer afford to ignore MRI and US as means to measure disease activity and joint damage in rheumatoid arthritis. PMID- 12737324 TI - Serum and salivary neopterin and interferon-gamma in primary Sjogren's syndrome. Correlation with clinical, laboratory and histopathologic features. AB - OBJECTIVE: To investigate serum and salivary neopterin and interferon-gamma as possible markers of immune system activation in primary Sjogren's Syndrome (pSS). METHODS: Serum and salivary neopterin and interferon-gamma concentrations were determined in 30 untreated patients with pSS and matched with several other clinical and laboratory parameters. RESULTS: The mean concentration of neopterin was significantly higher in pSS patients (8.12+/- 3.36 nmol/L in serum and 9.50 +/-7.61 nmol/L in saliva) than in normal controls (p<0.05). Significant correlations were found between serum neopterin and beta2-microglobulin, serum IgG as well as lip biopsy score. Salivary neopterin concentration was inversely related to Shirmer-I test, tear break-up time and stimulated salivary flow rate. Serum and salivary levels of interferon-gamma were normal and no correlation with the other parameters was found. CONCLUSION: In pSS patients serum neopterin may represent a useful marker of cell-mediated immunity. On the other hand, salivary neopterin seems to reflect theglandular damage. PMID- 12737325 TI - Local production of Ro/SSA and La/SSB autoantibodies in the target organ coincides with high levels of circulating antibodies in sera of patients with Sjogren's syndrome. AB - OBJECTIVE: To analyze Ro and La autoantibody levels in the periphery and production in the target organ in patients with Sjogren's syndrome. METHODS: The autoantibody production against the Ro and La proteins was investigated in 12 patients with Sjogren's syndrome. ELISA with recombinant antigens was used to determine levels in sera. The distribution of the antibody-producing plasma cells in the target organ was studied by immunohistochemistry using biotinylated antigens. RESULTS: All investigated patients with Sjogren's syndrome had detectable levels of Ro and La antibodies in sera, while local antibody production in the salivary glands was restricted to patients with high antibody levels. The autoantibody-producing cells were identified at the periphery of the infiltrates and in interstitial spaces. CONCLUSIONS: The chronic inflammation of the salivary glands in Sjogren's syndrome may be a self-sustaining process promoting autoantibody production in the target organ, possibly contributing to increased serum levels of autoantibodies. PMID- 12737326 TI - Reumacon (CPH82) showed similar x-ray progression and clinical effects as methotrexate in a two year comparative study on patients with early rheumatoid arthritis. AB - OBJECTIVES: To study x-ray development and clinical effects, tolerability and safety after 2 years treatment of RA patients with Reumacon (CPH82) or methotrexate (MTX). PATIENTS AND METHODS: This study is a 74 week open continuation of a 24 week double blind comparison of 100 patients with early RA (disease duration less than 2 years) treated either with Reumacon or MTX. RESULTS: The mean Larsen scores and the mean number of erosions increased significantly from baseline to 24 weeks and from 24 weeks to endpoint in both groups with no significant difference between them. Both groups had improved significantly in all clinical variables after 24 weeks and this improvement was sustained after two years. CONCLUSIONS: Radiological progression in patients treated with CPH82 was similar to that in patients treated with MTX. The clinical effect of the two drugs was sustained over the two year trial in both treatment groups. PMID- 12737327 TI - Outcome in adults with juvenile idiopathic arthritis. Comparison of the DUTCH AIMS2 between JIA and RA. AB - OBJECTIVES: To compare outcome in adult patients with JIA in childhood with outcome in young adults with RA and to evaluate the use of the DUTCH-AIMS2 in adult JIA patients METHODS: A questionnaire containing the DUTCH-AIMS2 to assess health outcomes (answers ranging from 0-10: 0=best) was sent to 142 adults with JIA (18-40 years). These health outcomes were compared with those of 34 young RA patients (< or = 40 years) from former studies. The DUTCH-AIMS2 was tested for its reliability and validity in adult JIA patients. RESULTS: The response rate was 71%. Disease duration of JIA patients (mean 19.2+/-9.5 years) was longer than in young RA patients (mean 7.8+/-5.3 years). RA patients scored worse than JIA patients in all health status areas with the exception of mobility. The mean health outcome scores were low for all groups (< or = 4.80). The internal consistency and the validity of the DUTCH-AIMS2 were satisfactory. CONCLUSIONS: Overall, the health outcome of oligoarticular and polyarticular adult JIA and young RA patients was quite good. The DUTCH-AIMS2 can be used as an instrument in outcome studies in JIA patients. PMID- 12737328 TI - Osteoclast formation from circulating precursors in osteoporosis. AB - OBJECTIVE: An imbalance between bone formation and bone resorption is thought to underlie the pathogenesis of reduced bone mass in osteoporosis. Bone resorption is carried out by osteoclasts. which are formed from marrow-derived cells that circulate in the monocyte fraction. The aim of this study was to determine the role of osteoclast formation in the pathogenesis of bone loss in osteoporosis. METHODS: The proportion of circulating osteoclast precursors and their relative sensitivity to the osteoclastogenic effects of M-CSF. 1,25(OH)2D3 and RANKL were assessed in primary osteoporosis patients and normal controls. RESULTS: Although there was no difference in the number of circulating osteoclast precursors in osteoporosis patients and normal controls. osteoclasts formed from osteoporosis patients exhibited substantially increased resorptive activity relative to normal controls. Although no increased sensitivity to the osteoclastogenic effects of 1,25(OH)D3 or M-CSF was noted, increased bone resorption was found in osteoporosis peripheral blood mononuclear cell (PBMC) cultures to which these factors were added. CONCLUSION: Our findings suggest that osteoclast functional activity rather than formation is increased in primary involutional osteoporosis and that dexamethasone acts to increase osteoclast formation. PMID- 12737329 TI - Reliability and validity of the swedish version of child health questionnaire. AB - Child Health Questionnaire (CHQ) is a North American self-administered instrument to assess physical and psychosocial health status for children (CHQ-CF) and their parents (CHQ-PF). We have determined reliability and validity of the CHQ on 45 children with JCA, who were treated as outpatients. Reliability (Cronbach's alpha) ranged between .86 and .94 for the dimensions of the CHQ-CF and between .69 and .94 for the CHQ-PF. Significant correlations (p<.01) were obtained between 7 of the CHQ dimensions and the Child Health Assessment Questionnaire (CHAQ) categories, and were particularly strong in the physical domain. Evidence of validity was found through significant correlations (p<0.01) between the CHQ and disease variables. Correlations between parents' and children's responses ranged between .366 and .786. CHQ was found to be valid and reliable for children with JCA and their parents. It seems particularly important to recommend that the children themselves are asked about their perceptions of health status and quality of life. PMID- 12737330 TI - Physiotherapy in subtropic climate improves functional capacity and health related quality of life in Swedish patients with rheumatoid arthritis and spondylarthropathies still after 6 months. AB - OBJECTIVE: The objective was to prospectively evaluate the short as well as the long-term effects of intensive physiotherapy in a stable, sunny and warm climate on physical function and health related quality of life in patients with rheumatoid arthritis (RA) and spondylarthropathies (SpA). METHODS: Ninety-three Swedish patients with RA and SpA receiving physiotherapy for 4 weeks in Israel or Tenerife were followed for 6 months. Physical function was evaluated by the Swedish version of Stanford Health Assessment Questionnaire (HAQ) and quality of life by the Nottingham Health Profile (NHP) questionnaire. RESULTS: There were significant improvements in HAQ-scores and global NHP-scores as well as all subcategories of NHP immediately after the treatment abroad, effects that were still measurable after six months. At that time point nearly half of the patients had clinically meaningful reduction of HAQ-scores (> or = 0.25). CONCLUSION: Physiotherapy in a warm and stable climate, with many hours of daily sunshine, is a valuable treatment complement for Swedish patients with RA and SpA. PMID- 12737331 TI - Effectiveness of low-level laser therapy in temporomandibular disorder. AB - OBJECTIVE: To investigate the effectiveness of low-level laser therapy in the treatment of temporomandibular disorder and to compare treatment effects in myogenic and arthrogenic cases. METHODS: Thirty-five patients were evaluated by magnetic resonance imaging and randomly allocated to active treatment (n=20) and placebo treatment (n= 15) groups. In addition to a daily exercise program, all patients were treated with fifteen sessions of low-level laser therapy. Pain, joint motion, number of joint sounds and tender points were assessed. RESULTS: Significant reduction in pain was observed in both active and placebo treatment groups. Active and passive maximum mouth opening, lateral motion, number of tender points were significantly improved only in the active treatment group. Treatment effects in myogenic and arthrogenic cases were similar. CONCLUSION: Low level laser therapy can be considered as an alternative physical modality in the management of temporomandibular disorder. PMID- 12737332 TI - Bone marrow findings in patients with adult Still's disease. AB - OBJECTIVE: Peripheral neutrophilia is one of characteristic laboratory findings in patients with adult Still's disease (ASD). We performed this study to identify the bone marrow findings in patients ASD. METHODS: We examined 12 bone marrow biopsy specimens from patients with ASD. RESULTS: The most frequent finding was granulocyte hyperplasia (12/12, 100%) and hypercellularity was observed in 75.0% (8/12) of patients. Plasmacytosis was present in 8.3% (1/12) of specimens. Histiocytosis and reactive hemophagocytosis were found in 25.0% (4/12) and in 16.7% (2/12) respectively. CONCLUSION: Our results show that bone marrow granulocyte hyperplasia is the main possible mechanism for peripheral neutrophilia and that histiocytic activation is a not infrequent bone marrow findings in patients with ASD. PMID- 12737333 TI - Treatment of reactive arthritis with infliximab. AB - We describe two patients with acute reactive arthritis (ReA) caused by Yersinia enterocolitica who were treated with infliximab with a good response in the acute phase. In a patient with prior ankylosing spondylitis (AS) and subsequent ReA the peripheral joint disease responded well to three infliximab infusions, but later he needed retreatment with infliximab to control the spinal symptoms of AS. The other previously healthy patient with acute ReA also responded well to one infliximab infusion although he did not fully recover and needed further treatment with disease modifying antirheumatic drugs. The influence of new drugs on the short and long term prognosis of the patients with ReA remains to be evaluated. PMID- 12737334 TI - Obstructive sleep apnea syndrome in children. AB - Obstructive sleep apnea syndrome (OSAS) in children is often caused by obstruction of the upper airway due to hypertrophy of the adenoids and palatine tonsils. Between October 1988 and December 1991, 50 children (34 males, 16 females) visited our department due to attacks of sleep apnea and underwent adenotomy or adeno-tonsillectomy. Respiratory monitoring during sleep was performed before and after operation, and the usefulness of the surgery was evaluated. Before operation, 27/ 50 children (54.0%) were diagnosed as having OSAS. Their age distribution showed peaks at the ages of 4 and 5 years and the male:female ratio was 2:1. Concerning the degree of improvement in clinical symptoms after surgery, marked effects were observed in 40/50 patients (80.0%), moderate effects in 7 (14.0%) and slight effects in 3 (6.0%). Concerning the degree of improvement in the apnea index after surgery in the 27 patients with OSAS, marked effects were observed in 22 patients (81.50%), moderate effects in 2 (7.4%), slight effects in 1 (3.7%) and no change in 2 (7.4%). PMID- 12737335 TI - Long-term surgical follow-up of sleep apnea syndrome. AB - During a 10-year period, 845 patients suspected of having sleep apnea syndrome underwent overnight monitoring at our institution. We report herein the results of a study of 204 surgical patients who responded to a questionnaire survey. Of these 204 cases, 86 (42%) showed a > or = 75% postoperative improvement in the apnea-hypopnea index (AHI) (or AHI < 10) and 48 (24%) showed an improvement of > or = 50%. Based on the data obtained from the questionnaire survey we studied the long-term postoperative improvement as well as the long-term prognosis in terms of the postoperative improvements in AHI and body weight. PMID- 12737336 TI - Obesity and obstructive sleep apnea syndrome. AB - We investigated the influence of obesity on upper airway obstruction, especially the relationship between obesity and the type of obstruction. The site of obstruction was identified by means of endoscopic examination and dynamic MRI during sleep. Many obese patients have the circumferential type of obstruction. PMID- 12737337 TI - Anatomy of oral respiration: morphology of the oral cavity and pharynx. AB - The anatomical states of the oral cavity and pharynx during mouth breathing in children with adenoid hypertrophy and in adults confirmed the speculation that mouth breathing is disadvantageous compared with nose breathing. In addition, comparison of the anatomical state between wakefulness and sleep in normal adults showed slight depression of the tongue root and slight narrowing of the oropharynx and hypopharynx during sleep. Obstructive sleep-disordered breathing occurs due to a variety of factors, such as paranasal sinus disease, tonsil and adenoid hypertrophy, hypertrophy and morphological abnormalities of the soft palate and palatine uvula, low-set soft palate, micrognathia, macroglossia, obesity and tongue root depression. Narrowing or obstruction of the middle pharynx and hypopharynx is more marked in patients with obstructive sleep disordered breathing than in normal people and is especially marked during sleep. Therefore, morphological (i.e. anatomical) changes during mouth breathing may provide useful information for evaluating the pathology of snoring and sleep apnea. PMID- 12737338 TI - Dynamic MRI diagnosis of sleep-disordered breathing. AB - According to the obstructive site, the different types of obstructive-type sleep disordered breathing are classified as the soft palate type, palatine tonsil type, soft palate and depressed lingual root type, depressed lingual root type and epiglottis type. In this study, we conducted dynamic MRI examinations of the oral, pharyngeal and lingual root sites in patients with obstructive sleep disordered breathing, and studied the usefulness of MRI as a diagnostic tool for the examination of such obstructive sites. PMID- 12737339 TI - Influence of sleep respiratory disturbance on nocturnal blood pressure. AB - We subjected 65 patients to overnight monitoring and continuous nocturnal blood pressure measurement in order to assess the influence of sleep apnea on the circulatory system. Thirty-one patients were compared before and after surgery. The severity of sleep apnea was classified by means of the apnea-hypopnea index (AHI), the duration of exposure to low-level oxygen [calculated as the desaturation time (DT)] and changes in blood pressure (BP). Before surgery, a significant correlation was noted between DT and changes in BP. Therefore, DT was considered useful for assessing the influence of sleep apnea on nocturnal BP. After surgery, an improvement in AHI of > 50% was noted in 19/31 patients (61.3%), a result comparable to that described in the literature. Improvements in DT and change in BP were > 50% in 21/31 (67.7%) and 14/ 31 (45.2%) patients, respectively. With regard to the severity of sleep apnea before surgery, AHI was > or = 50 and DT > or = 40% in 10 and 18 patients, respectively and 19 patients showed changes in BP of > or = 40 mmHg. After surgery, one, five and two patients, respectively, still showed these values. Thus, a beneficial effect of surgery was demonstrated. PMID- 12737340 TI - Endoscopic examination of obstructive sleep apnea syndrome patients during drug induced sleep. AB - Sixty patients diagnosed with obstructive sleep apnea syndrome (OSAS) underwent uvulopalatopharyngoplasty (UPPP). The effects of surgery were studied based on endoscopic findings during drug-induced sleep and determination of the apnea hypopnea index (AHI) before and after the operation. Changes in the form of the airway during sleep in the recumbent position were observed, and the role of upper airway endoscopy in the diagnosis and surgical treatment of OSAS was determined. The site of airway obstruction during sleep induced by i.v. injection of 10 mg of diazepam was classified into five types, and changes in AHI and the site of airway obstruction were compared before and after surgery. Changes in airway morphology during sleep in the supine and recumbent positions were also compared before surgery. The postoperative improvement rate was 74.4% for the soft palatal type of obstruction, 76.2% for the tonsillar type, 53.3% for the circumferential palatal type and 34.0% for the mixed type. Treatment produced excellent or good effects for the soft palatal and tonsillar types of obstruction. However, many patients with the circumferential palatal and mixed types of obstruction showed only some improvement or no change. Good airway morphology was maintained in the recumbent position by patients with the soft palatal type of obstruction. With the circumferential palatal and mixed types of obstruction, improvement can be expected from operations which include surgical treatment of the posterior pharyngeal wall or lateral funiculus, or with midline laser glossectomy. A good operative outcome can be predicted in patients showing improvement of apnea in the recumbent position preoperatively. PMID- 12737341 TI - Posture of patients with sleep apnea during sleep. AB - The relationship between sleep apnea syndrome (SAS) and posture during sleep has been noted and the beneficial effect of an optimal posture on sleep apnea has been empirically indicated. We investigated this effect in a group of subjects that included obese patients and found that the apnea-hypopnea index (AHI) may be normalized in the lateral position, even among patients severely affected with apnea. Among those with intermediate or lower AHI values sleeping in a lateral position markedly improved the symptoms, with AHI even approaching the normal range in many patients. A tendency was noted for AHI to rise regardless of posture but in proportion to the increase in body mass index (BMI). In other words, the improvement due to changes in posture became increasingly insignificant with increase in BMI. PMID- 12737343 TI - Comparison of the state of sleep in patients with sleep-disordered breathing before and after surgical treatment. AB - In 19 surgically treated patients with sleep-disordered breathing for whom electroencephalograms could be recorded before and after surgery, and in 10 patients who suffered from daytime lethargy as assessed using the Epworth Sleepiness Scale (ESS) score, correlations between the stages of sleep before surgery and the ESS score and apnea-hypopnea index (AHI) were evaluated, and changes in AHI and the stages of sleep after surgery were examined. Neither the preoperative severity of AHI nor the ESS score showed an association with the stages of sleep. The improvement in the quality of sleep tended to be greater as the percentage improvement in the ESS score increased. PMID- 12737342 TI - Comparison of surgery and nasal continuous positive airway pressure treatment for obstructive sleep apnea syndrome. AB - Uvulopalatopharyngoplasty (UPPP) and nasal continuous positive airway pressure (n CPAP) are used for the treatment of obstructive sleep apnea syndrome (OSAS). Although OSAS results from an abnormality of the pharynx, very little research has been carried out regarding the selection of UPPP or n-CPAP according to the type of abnormality. We performed n-CPAP titration before and after surgery, compared the treatment methods and evaluated the effect of the medical therapy. A sleep polygraph was recorded on the first night of admission and patients diagnosed with OSAS underwent n-CPAP titration on the second night. The blocked region was identified by means of endoscopic examination. A couple of months after surgery, polysomnography was repeated to determine the effect of surgery and the pressure level during n-CPAP forpatients who showed < 50% improvement in the apnea-hypopnea index. The results of surgery were poor in cases revealed by endoscopy to have the circumferential type of obstruction, but good for the soft palate and tonsil types. When endoscopic examinations were performed in conjunction with n-CPAP, the treatment was observed to act on the pharynx and to expand the airway in all cases Combined medical treatments were effective in cases where n-CPAP alone was ineffective due to high pressure. PMID- 12737344 TI - Laser midline glossectomy and lingual tonsillectomy as treatments for sleep apnea syndrome. AB - Preservation treatments for sleep respiratory disorders, such as the use of a dental device and the technique of nasal continuous positive air pressure, cause discomfort to the patient and are not radical treatments. Therefore, we performed operative therapy instead. Laser midline glossectomy was performed to treat constriction at the root of the tongue in 16 patients diagnosed with sleep apnea syndrome. We also tried lingual tonsil excision using the Harmonic Scalpel in three patients with stenosis at the base of the tongue. PMID- 12737345 TI - Two cases of sleep apnea syndrome caused by primary hypothyroidism. AB - Two cases of sleep apnea syndrome caused by primary hypothyroidism are reported. The first patient was a 66-year-old man who complained of sleep apnea; his apnea hypopnea index (AHI) was 50.8, as assessed by all-night monitoring. Hypothyroidism was subsequently suspected when he showed delayed recovery from general anesthesia following surgery involving uvulopalatopharyngoplasty. Hypothyroidism was diagnosed on the basis of blood tests His snoring and apnea improved after 2 months of levothyroxine sodium administration and the AHI fell from 50.8 to 13.0. The second patient was a 73-year-old man with an AHI of 41.3, as assessed by all-night monitoring. Hypothyroidism was diagnosed on the basis of blood tests and was suspected because of his slow speech. He was similarly treated with levothyroxine sodium. The AHI did not decrease after 4 months of treatment. His desaturation rate (rate of O2 saturation < 90%) improved however, from 56.6% to 31.9%, and the symptoms of hypothyroidism also improved markedly. In both patients, elevated creatine phosphokinase, a dull facial expression, peripheral edema and slow speech were recognized, and these symptoms were suggestive of hypothyroidism. The type of sleep apnea was mainly obstructive in both patients. PMID- 12737346 TI - Application of lingual tonsillectomy to sleep apnea syndrome involving lingual tonsils. AB - In sleep apnea syndrome, surgical treatment is applied in obstructive-type cases and some mixed-type cases. If the obstructive part is in the root of the tongue, forward transfer of the tongue, lingual tonsillectomy and laser midline glossectomy are applied. In this study, we demonstrate the surgical technique of lingual tonsillectomy using an ultrasonic coagulating dissector (SonoSurg) with a blade tip shape developed in our department. We conclude that lingual tonsillectomy using SonoSurg, which we have frequently used, should be the first choice of treatment for snoring and sleep apnea caused by hypertrophy of the lingual tonsils from the viewpoints of effectiveness, prevention of hemorrhage, safety and handling. PMID- 12737347 TI - Pathophysiology of nerve injury. AB - The response to nerve injury is a complex and often poorly understood mechanism. An in-depth and current command of the relevant neuroanatomy, classifications systems, and responses to injury and regeneration are critical to current clinical success. Continued progress must be made in our current understanding of these varied physiologic mechanisms of neuro-regeneration if any significant progress in clinical treatments or outcome is to be expected in the future. Reconstructive surgeons have in many ways maximized the technical aspects of peripheral nerve repair. However, advances in functional recovery may be seen with improvements in sensory and motor rehabilitation after peripheral nerve surgery and with a combined understanding of the neurobiology and neurophysiology of nerve injury and regeneration. PMID- 12737348 TI - Evaluation of the nerve-injured patient. AB - The evaluation of patients with nerve injury or nerve compression requires an accurate history and subjective report to determine the tests that are the most useful in providing the essential information. Motor and sensory evaluation is necessary inglobal mixed-nerve injuries, but in cases of nerve compression, tests of provocation give more accurate information for detecting the site of nerve compression. There is no gold standard test in the evaluation of patients with nerve injury or compression; therefore, a battery of valid and reliable sensory and motor tests provides the most complete information to formulate a treatment plan. PMID- 12737349 TI - The electrodiagnostic examination with peripheral nerve injuries. AB - The EDX examination can be of considerable use to the clinician in the evaluation of focal peripheral nerve injuries. It can confirm the presence ofsuch lesions and help determine their location, severity, and prognosis. The major reason why it is capable of doing so is because it discloses the pathophysiology of focal neuropathies, at least in regard to large myelinated nerve fiber damage. PMID- 12737350 TI - Compressive neuropathies of the upper extremity. AB - Although any peripheral nerve may be compressed anywhere along its course, nerve compression syndromes typically occur at predictable sites with predictable clinical presentations. A detailed history and physical examination can establish a diagnosis, and electrodiagnostic studies and at times imaging can confirm it. Physicians should adopt a systematic approach of diagnosing and treating these types of nerve lesions. The physician and the patient can be rewarded with favorable outcomes. PMID- 12737351 TI - Thoracic outlet syndrome. AB - Success in conservative management depends upon an accurate assessment and development of a treatment plan relative to the irritability of the patient's condition. Postural correction and avoidance of irritating positions must begin early in the rehabilitation phase to retrain the patient in symptom-reducing postures. Treatment addressing only the neurovascular structures may produce temporary relief of symptoms, but postural correction cannot be maintained without correction of the associated muscle imbalance in the cervicoscapular region. Long-term success of conservative management depends on patient compliance to a home exercise program and behavior modification at home and at work. Surgical decompression should be reserved for patients who fail to improve with conservative management. PMID- 12737352 TI - Compression neuropathies of the lower extremity. AB - Compression neuropathies of the lower extremity can be just as disabling as compression neuropathies in the upper extremity. The most common compression neuropathies encountered in the lower extremity affect the lateral femoral cutaneous nerve, the peroneal nerve, the saphenous nerve, the sural nerve, and the tibial nerve (including their branches). These are discussed in terms of anatomy, etiologies, manifestations, and surgical treatment. PMID- 12737353 TI - Nerve repair, grafting, and nerve transfers. AB - Advances in the field of peripheral nerve surgery have increased our understanding of the complex cellular and molecular events involved in nerve injury and repair. Application of these important discoveries has led to important developments in the techniques of nerve repair, nerve grafting, nerve allografts, end-to-side repairs, and nerve-to-nerve transfers. As our understanding of this dynamic field increases, further improvement in functional outcomes after nerve injury and repair can be expected. PMID- 12737354 TI - Tendon transfers for failed nerve reconstruction. AB - There are few reconstructive procedures in the upper extremity that are as helpful to patients as a tendon transfer. Successful tendon transfer requires the marriage of anatomic knowledge, surgical judgment, and rehabilitative expertise. Frequently, the improvement in function can be dramatic. By the same token, an unsuccessful tendon transfer wastes a normal muscle-tendon unit in the hand and leaves the patient withless function than what was present preoperatively. PMID- 12737356 TI - Brachial plexus injuries. AB - Severe trauma to the brachial plexus most often occurs in young adult men and is a crippling injury that requires management in a timely fashion for optimal functional recovery and pain control. The surgical management of such injuries is well established, and the techniques continue to evolve. Current management options consist of primary repair in the acute setting, neurolysis, neuroma resection and nerve grafting, motor and sensory nerve transfers, and muscle and tendon transfers. Shoulder andwrist fusion can also play a role in the overall management of these patients. The best operative plan varies depending on the patient's level and extent of injury and the surgeon's preference and experience. The pre- and postoperative care of these patients is ideally managed by a team that has experience with such problems, including personnel knowledgeable in their postoperative rehabilitation. The total reconstructive process generally consists of more than one operation, and the postoperative rehabilitation is long and intensive. Nevertheless, with a highly motivated patient and a dedicated and specialized surgical team, the prognosis for functional recovery is good, and these patients can still lead productive and satisfying lives. PMID- 12737355 TI - Management of neuromas. AB - After nerve injury and regeneration, significant pain may be associated with the scar and altered sensation observed within the distribution of the injured nerve. A bulbous swelling may form at the severed nerve end, constituting a traumatic neuroma. The development of a painful neuroma may be more disabling to the patient than an area of anesthesia or even loss of motor function. Effective treatment of the painful neuroma remains a difficult problem. Diminished productivity, alterations in patient lifestyle, and possible progression to chronic pain syndromes must be considered within the scope of neuroma management, and treatment must focus on alleviating the pain and restoring the functional loss caused by the nerve injury. Careful patient selection is the cornerstone of successful outcomes. Once the patient has been selected, the surgical management of the painful neuroma throughout the body is based on basic principles that vary only slightly from region to region. Using these tenets, a neuroma management algorithm has been developed based on the pathophysio-logy of the neuroma, the results of experimental studies, review of patient outcomes, and understanding the psychology of pain in the surgical patient. PMID- 12737357 TI - Management of obstetrical brachial plexus palsy evaluation, prognosis, and primary surgical treatment. AB - Primary surgery for obstetrical brachial plexus lesions is a young field of surgical expertise that offers the possibility of improved functional ability in carefully selected patients who would otherwise be faced with lifelong impairment and secondary skeletal deformities. One major challenge in this area of peripheral nerve surgery is the selection of patients most likely to derive benefit from surgical intervention. The key to the development of selection criteria and to the resolution of other considerations (such as the determination of root avulsion) is consistency, accuracy, and careful reporting of natural history and outcome data. In particular, we strongly feel that a statistically sound technique of assessment must be consistently applied from the time of presentation through long-term follow-up. Advancement to date has resulted from the application of evidence-based recommendations from large, well-designed, meticulous studies. As the field of obstetrical brachial plexopathy management continues to evolve, we can expect that questions will continue to be answered using such scientific methodology. PMID- 12737358 TI - The surgical management of facial nerve injury. AB - Treatment of facial nerve injuries depends upon a detailed understanding of its anatomic course, accurate clinical examination, and timely and appropriate diagnostic studies. Reconstruction depends upon the extent of injury, the availability of the proximal stump. and the time since injury and duration of muscle denervation. Although no alternative is perfect, these techniques, in combination with static and ancillary procedures. can protect the eye, prevent drooling, restore the smile, and improve facial symmetry. New techniques (including single-stage free tissue transfers and bioengineered nerve grafts), further research on the characteristics of the facial musculature, and methods of preserving the neuromuscular junction will undoubtedly manifest themselves as further refinements of established surgical techniques. PMID- 12737359 TI - The ceaseless pursuit of triumph over death in life. PMID- 12737360 TI - Why refer to a psychiatrist? PMID- 12737361 TI - Dying with dignity. PMID- 12737362 TI - Nuclear medicine and the physician. PMID- 12737363 TI - Poisons. Initial assessment and management. PMID- 12737364 TI - Self-poisoning in the UK: epidemiology and toxidromes. PMID- 12737365 TI - Antidepressant poisoning. PMID- 12737366 TI - Non-opioid analgesic poisoning. PMID- 12737367 TI - Acute effects of drugs of abuse. PMID- 12737368 TI - Cardioactive drugs. PMID- 12737369 TI - Assessing the performance of specialist registrars. AB - Assessing the performance of doctors while they are engaged in clinical work is a challenging concept. The introduction of objective-based curricula provides the stimulus and opportunity for the Royal Colleges of Physicians to develop relevant and reliable methods of in-service assessment. We propose to pilot a study investigating the validity, reliability and feasibility of three assessment methods--direct observation of the clinical encounter using an adapted mini-CEX, direct observation of the performance of practical procedures (DOPS), and the doctor's ability to perform effectively as part of a team using 360 degree assessment. The methods will be studied in the setting of routine clinical care. Whilst demanding of time from both trainees and trainers, they will represent a significant advance on the current system which is characterised by a lack of evidence in the assessment process. PMID- 12737370 TI - The development of clinical governance in the UK: its implications for rehabilitation medicine. AB - Clinical governance has been introduced in the UK as a mechanism to improve the quality of health services, and the professional performance of clinicians. Specialist societies have a pivotal role to play in developing systems to support its implementation in clinical practice. Rehabilitation medicine is a specialty characterised by closely integrated interdisciplinary teamwork. The particular issues that this poses for clinical governance may also be relevant to other specialities. This paper presents an overview of the development of audit and clinical governance procedures in the UK and their specific implications for rehabilitation medicine. It also summarises the work undertaken to date by the British Society for Rehabilitation Medicine towards implementation of clinical governance, to improve the quality of care offered to patients. PMID- 12737371 TI - A pretty pass: when is there a right to die? AB - This paper discusses the present extent and limits of a persons right to choose when and how to die, in the light of the contrasting decisions in the cases of Mrs Pretty (who was not allowed the choice) and Ms B (who was), and of the European Convention of Human Rights. It also discusses whether the Convention might eventually develop a right to self determination which would include such a right. PMID- 12737372 TI - 'If only someone had told me . . .' A review of the care of patients dying in hospital. AB - Approximately half of all patients who die do so in hospital. Despite the advent of palliative care in the UK, there is evidence that the care that many patients receive in the final phase of their illness in hospital is poor. Building on a study of bereaved relatives' views of the information provided by an inner city hospital trust during an admission in which a patient died, this article explores the factors that may contribute to sub-optimal care for patients dying in hospital. In particular, a lack of open communication, difficulties in accurate prognostication and a lack of planning of end-of-life care can all result in poor care. Strategies to improve care, such as the use of integrated care pathways, advance directives and education initiatives, are discussed. PMID- 12737373 TI - The natural history and management of hepatorenal disorders: from pre-ascites to hepatorenal syndrome. AB - In cirrhosis, the natural history of hepatorenal disorders starts with a pre ascitic stage and is followed by the development of ascites; hepatorenal syndrome (HRS) begins with compensated renal sodium retention, or pre-ascites. In pre ascites, the renal sodium retaining tendency leads to 'overfilling' of total blood volume, with increased glomerular filtration rates (GFR), overcoming the renal sodium retaining tendency possibly due to renal accumulation of angiotensin II. As peripheral vasodilatation increases, the vascular capacity (in effect the arterial blood volume) becomes inadequately filled, GFR falls, compensatory vasoconstrictors rise, and the resulting renal sodium retention results in diuretic-responsive ascites formation. Increasing proximal reabsorption of sodium results in ascites refractory to diuretic therapy. Repeated abdominal paracentesis will not prevent insidious progression to HRS type II, nor to the precipitation of HRS type I. In contrast, liver transplantation, or transjugular intrahepatic hepatoportal stent shunt (TIPS) in refractory ascites, may prevent the onset of, or reverse, HRS. However, recent non-controlled studies indicate exciting possibilities of medical therapy reversing HRS. PMID- 12737374 TI - Treatment in art. PMID- 12737375 TI - Liver cells: biology to therapeutics. AB - The liver has a remarkable capacity to regenerate after injury. Recent work has demonstrated that repair may call upon either the division of pre-existing mature cells, or the expansion of intrahepatic progenitor cells. Furthermore, progenitors may migrate into the liver from the bone marrow. Understanding and exploiting the cell biology of the liver provides the basis for innovational treatment, including the use of growth factors, transplantation of isolated cells, genetic manipulation of hepatocytes and liver cell progenitors, and the development of artificial liver support systems. PMID- 12737376 TI - Recent advances in coeliac disease. AB - Coeliac disease, or gluten-sensitive enteropathy, affects 1 in 100-200 people in the UK. The condition, which is exacerbated by wheat, rye, barley and possibly oats, can be treated with a gluten-free diet in which these cereals are omitted. Serological screening, particularly of high-risk groups, with both IgA and IgG based systems can be used to identify cases. Diagnosis depends on the use of a small intestinal biopsy, which reveals the classical changes of loss of the normal villous architecture. Evidence suggests that gluten-sensitive T cells are involved in the pathogenesis of the disease. Use of in vitro systems has suggested an immunodominant epitope within wheat gliadin, which has been shown to exacerbate the condition in vivo. This information can be used to devise strategies to develop immuno-modulatory peptides and cereals with the baking and nutritional qualities of wheat, rye and barley, but which do not exacerbate the condition. PMID- 12737377 TI - The psychological care of medical patients. PMID- 12737378 TI - Recent advances in predicting the response to clinical rehabilitation. PMID- 12737379 TI - Artificial nutrition and hydration: science, ethics and law. PMID- 12737380 TI - Axel Munthe's The story of San Michele revisited. AB - This article provides a review of Axel Munthe's best-selling medical novel, The Story of San Michele, with particular reference to the light it sheds on late nineteenth and early twentieth century medicine. PMID- 12737381 TI - The use of oxygen in acute exacerbations of chronic obstructive pulmonary disease: a prospective audit of pre-hospital and hospital emergency management. PMID- 12737382 TI - The use of oxygen in acute exacerbations of chronic obstructive pulmonary disease: a prospective audit of pre-hospital and hospital emergency management. PMID- 12737383 TI - The use of oxygen in acute exacerbations of chronic obstructive pulmonary disease: a prospective audit of pre-hospital and hospital emergency management. PMID- 12737384 TI - The use of oxygen in acute exacerbations of chronic obstructive pulmonary disease: a prospective audit of pre-hospital and hospital emergency management. PMID- 12737385 TI - Cultural differences: practising medicine in an Islamic country. PMID- 12737386 TI - Conversations with Charles. PMID- 12737387 TI - Driving restrictions after stroke: doctors' awareness of DVLA guidelines and advice given to patients. PMID- 12737388 TI - Driving restrictions after stroke: doctors' awareness of DVLA guidelines and advice given to patients. PMID- 12737389 TI - Consent with understanding: a movement towards informed decisions. PMID- 12737390 TI - Healthy limb amputation: ethical and legal aspects. PMID- 12737391 TI - AIDS and CFS/ME. PMID- 12737392 TI - Diagnostic error in the hospital presentation of acute asthma. PMID- 12737393 TI - Availability of fully staffed GI endoscopy lists at the weekend for inpatients: does it make a difference? PMID- 12737394 TI - Microarray analysis in the clinical management of cancer. AB - Because of the many genetic and epigenetic alterations that define a given tumor cell, and given the heterogeneity of these changes, between, as well as within a tumor class, progress in the understanding and treatment of cancer has been slow. The sequencing of the human genome, in combination with advances in robotics, computing, and imaging technologies, has resulted in rapid advances in the development of microarray methodology. This technology now places us in a position to simultaneously consider the consequence of all of these genetic changes through measurement of a large proportion of the complement of genes expressed in a given tissue at a given time. The power of this methodology for (1) the classification and identification of tumor classes, (2) gene discovery, (3) determining mechanisms of drug action, and (4) predicting drug response has now clearly been demonstrated: however, several challenges remain. First, there is a growing need for a standardization of the methodology, such that different datasets may be compared directly and meaningfully. The complete sequencing and annotation of the human genome may be the first step toward this attainable goal. Also, the databases generated should be made publicly available to facilitate further analysis by other researchers. To this end, the results of our studies are available at http://sequence.aecom.yu.edu/bioinf/Augenlicht/default.html. Second, a number of reported findings on tumor classification require further validation in independent patient data sets. Third, extensive clinical studies with appropriate patient follow-up are required to determine the validity of this method for the prediction of patient response to chemotherapy. Finally, the possibility needs to be considered that gene expression profiling may need to be combined with other global approaches such as proteomics and mutation screening analyses, for optimization of its potential. The advent of methodologies that enable gene expression profiling provides an opportunity to gain insights into the genetic makeup of a cancer cell on a global scale. Given the heterogeneity of this disease, such a global approach is likely to enhance significantly our understanding and management of this disease. PMID- 12737395 TI - Quassinoids from Picrasma crenata. AB - From woods of Picrasma crenata, a new stereoisomer dihydronorneoquassin was obtained together with others well knowns dihydronorneoquassin, parain, alpha neoquassin, beta-neoquassin and quassin. The structures were determined by spectroscopic data and chemical evidence. PMID- 12737396 TI - A new spiroketal steroid from Gorgonella umbraculum. AB - A new spiroketal steroid (2) was isolated from the Indian Ocean gorgonian, Gorgonella umbraculum besides the known briarane diterpenoids, juncellin, umbraculides A and D. The new steroid was characterised as 22-acetoxy-3,25 dihydroxy-16-24,20-24-bisepoxy-(3beta, 16alpha, 20S, 22R, 24S)cholest-5-ene (2) by a study of its spectral data. PMID- 12737397 TI - Triterpenoidal constituents of the leaves of Carissa carandas. AB - Studies undertaken on the fresh leaves of Carissa carandas have led to the isolation of four pentacyclic triterpenoids (1-4) including one new constituent carissin (1) and two hitherto unreported compounds 2 and 3. The structure of the new triterpenoid has been elucidated as 3beta-hydroxy-27-E-feruloyloxyurs-12-en 28-oic acid. Compelete assignment of the protons of 2 has also been made based on 2D NMR studies. PMID- 12737398 TI - Chemistry and biological significance of essential oils of Cymbopogon citratus from Pakistan. AB - Steam distilled oil of Cymbopogon citratus was analyzed by Gas Chromatographic Mass Spectrometry (GC-MS) and citral was found as major constituent. The oil exhibited significant inhibition of beta-glucuronidase activity and also showed activities against some tested human, plant and animal pathogens. The minimum inhibitory concentrations could not be determined due to the lack of some chemicals. PMID- 12737399 TI - Anti-inflammatory activity of a novel flavonol glycoside from the Bauhinia variegata Linn. AB - Bauhinia variegata Linn. (Leguminosae) is commonly known as 'Kachnar' in Hindi. It is distributed almost throught India. Its powdered bark is traditionally used for tonic, astrain, ulcers. It is also useful in skin diseases. The roots are used as antidote to snake poison. The present article deals with the isolation and structural elucidation of a novel flavonol glycoside 5,7,3',4'-tetrahydroxy-3 methoxy-7-O-alpha-L-rhamnopyranosyl(1-->3)-O-beta-galactopyranoside (1) from the roots of Bauhinia Variegata and its structure was identified by spectral analysis and chemical degradations. The novel compound (1) showed anti-inflammatory activity. PMID- 12737400 TI - Isolation and spectroscopic study of pectic substances from kenaf (Hibiscus cannabinus L.). AB - Pectins were isolated from bark, wood and pith of kenaf (Hibiscus cannabinus L.) variety Cuba108. Imidazole was used as extractant. The isolated pectins were studied by 13C nuclear magnetic resonance (13C-NMR) spectroscopy and diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS). From the spectroscopic investigation, the isolated pectic substances were found similar to commercial pectins. PMID- 12737401 TI - Synthesis of degraded cyanogenic glycosides from Sambucus nigra. AB - Two natural cyanohydrins, isolated from Sambucus nigra, have been synthesised from mandelonitrile and penta-O-acetyl-beta-galactopyranose. The synthesis confirmed the stereochemistry of the compounds, which had been assigned on biogenetic grounds. PMID- 12737402 TI - Glycosides from the leaves of Ilex hylonoma. AB - Two new triterpenoid saponins, hylonosides I-II (1,2) have been isolated along with one known flavonoid glycoside (3) from the methanol extract of leaves of Ilex hylonoma. The structures were elucidated by ID and 2D NMR experiments, including 1H-1H COSY, TOCSY, NOESY, HMQC, and HMBC methods. Two new compounds were characterized as 3-O-beta-D-glucopyranosyl-(1 --> 4)-beta-D glucuronopyranosyl pomolic acid-28-O-beta-D-glucopyranoside (1), 3-O-beta-D glucopyranosyl-(1 --> 4)-beta-D-glucopyranosyl-(1 --> 2)-beta-D glucuronopyranosyl pomolic acid-28-O-beta-D-glucopyranoside (2). PMID- 12737404 TI - First 2-hydroxy-3-methylbut-3-enyl substituted xanthones isolated from plants: structure elucidation, synthesis and antifungal activity. AB - Two new 2-hydroxy-3-methylbut-3-enyl substituted xanthones, (+/-)-caledol 1 and (+/-)-dicaledol 2 were isolated from a dichloromethane extract of the leaves of Calophyllum caledonicum (Clusiaceae). Compounds 1 and 2 are the first 2-hydroxy-3 methylbut-3-enyl substituted xanthones isolated from natural source. Their structures were elucidated by means of combined analytical methods including HRFABMS, 1D and 2D NMR spectroscopies and also confirmed by total synthesis using biomimetic ortho-prenylphenols photooxygenation (1O2) as a key step. The antifungal activity against Aspergillus fumigatus is reported. PMID- 12737403 TI - GC-MS analysis and bioactivity testing of the volatile oil from the leaves of the toothbrush tree Salvadora persica L. AB - Upon GC-MS analysis of the volatile oil extracted from Salvadora persica L. leaves, we were able to identify benzyl nitrile, eugenol, thymol, isothymol, eucalyptol, isoterpinolene, and beta-caryophyllene. Toxicity of the aroma was evaluated using brine shrimp lethality test which gave an LC50 > 1,000 ppm. Using Disc Diffusion Test, it was found that the extract of the leaves has a considerable antibacterial effect on several different oral aerobic bacteria with comparable results to known antibiotics. The extract can be used effectively as a natural tool for teeth cleaning and as a natural analgesic for the disturbing toothache. PMID- 12737405 TI - New fluorenone and phenanthrene derivatives from Dendrobium chrysanthum. AB - Two new fluorenone derivatives denchrysans A (1) and B (2) and one new phenanthrenediglycoside denchryside A (3) were isolated from the herbs of Dendrobium chrysanthum (Orchidaceae). Their structures were established on the basis of spectral evidence. PMID- 12737406 TI - Synthesis and antimicrobial activities of some isocoumarin and dihydroisocoumarin derivatives. AB - A new series of the isocoumarin derivatives of ibuprofen, fluribiprofen, naproxen, valproic acid and 1-naphthoic acid have been synthesized via condensation of homophthalic acid with their respective acid chlorides. The conversion of the latter two isocoumarins into (dl)-3,4-dihydroisocoumarins has also been achieved. Most of the synthesized compounds showed antifungal and antibacterial activities. PMID- 12737407 TI - Microbial transformation of dehydroepiandrosterone. AB - Transformation of dehydroepiandrosterone (DHEA) (1) was carried out by a plant pathogen Rhizopus stolonifer, which resulted in the production of seven metabolites. These metabolites were identified as 3beta,17beta-dihydroxyanandrost 5-ene (2), 3beta,17beta-dihydroxyandrost-4ene (3), 17beta-hydroxyandrost-4-ene-3 one (4), 3beta,11-dihydroxyandrost-4-ene-17-one (5), 3beta,7alpha-dihydroandrost 5-ene-17-one (6), 3A,7alpha,17beta-trihydroxyandrost-5-ene (7) and 11beta hydroxyandrost-4,6-diene-3,17-dione (8). The structures of the transformed products were determined by the spectroscopic techniques. PMID- 12737408 TI - Preparing for Pacific prosperity or disaster. PMID- 12737409 TI - Pacific response and disaster readiness. PMID- 12737410 TI - Pilot workshops at the Palau Center for Emergency Health: a model for international collaborative operations training and planning. AB - The Emergency Public Health Planning Workshop demonstrated that national with different backgrounds, capabilities, knowledge bases, and concepts of operations can work together to develop complementary emergency public health plans. Results of the First Responder Emergency Medical Workshop demonstrated that, despite having similarly inconsistent backgrounds, emergency responders can develop complementary response protocols. Lectures presented in both workshops are now available in the public domain. They can be used to improve regional public health emergency capabilities whether the region considered is local, state, or international. PMID- 12737411 TI - Assessing physicians' continuing medical education (CME) needs in the U.S. associated Pacific jurisdictions. AB - OBJECTIVE: To assess the self-perceived continuing medical education (CME) needs of physicians in American Samoa, Commonwealth of the Northern Mariana Islands, Guam, Federated States of Micronesia, Republic of the Marshall Islands, and the Republic of Palau. METHODS: Questionnaire-based survey of all physicians. RESULTS: Responses obtained from a total of 143 physicians in the region provided information on training backgrounds, previous experiences with CME, local access to regular CME sessions, perceived priority educational needs and preferred methods of CME delivery. CONCLUSIONS: Overall 64% of respondents had attended a formal CME event in 1999 or 2000, and 71% had access to local weekly or biweekly CME. However the perceived usefulness of these events varied by region. Priority learning needs were identified by physicians including non-communicable diseases such as diabetes, hypertension, cardiac disease; communicable diseases such as tuberculosis, HIV/AIDS and tropical diseases; as well as skills such as EKG and X ray interpretation, trauma management and cardiac life support. Information on the most pressing educational needs and desired methods of delivery will be crucial in planning CME in this region. PMID- 12737412 TI - Population, public health and tubal ligation in Vanuatu. AB - The association between population growth and public health is often neglected. Between 1989 and 1999 Vanuatu's population grew by 30%. Unless this growth slows existing public health problems will worsen. To gain insight into family planning habits 111 ni-Vanuatu women undergoing tubal ligation were surveyed. 85% had 4, or more, children. Women resident on the outer islands had more children than those on the island containing the nation's capital. The implication is that ni Vanuatu women perceive 4 children as desirable. With approximately half the population entering their reproductive years in the next decade this is too many to avoid a population explosion in Vanuatu. Public health campaigns must promote the benefits of a small family. Urgent education and widespread provision of effective family planning is required to avoid a future public health crisis. Aid donors need to be made aware of this as an absolute priority. PMID- 12737413 TI - Consultation-liason psychiatry in Fiji. AB - OBJECTIVE: To examine and report the characteristics of the patients seen on the consultation-liaison (C-L) psychiatry in the main general hospital in Fiji Islands. METHOD: The socio-demographic and clinical indices of all the patients referred to the C-L psychiatric service between January 1999 and June 2000 were retrieved from the computerized C-L psychiatry register and analyzed. These were compared, in certain aspects, with those of the patients seen in psychiatric hospital located within the same city. RESULTS: The rate of referral to the C-L service was 0.4% of all hospital admission and outpatient referrals. A major reason for referral was psychiatric comorbidity (80.5%). Other reasons include social problems, neurological conditions such as epilepsy and migraine. About fifty-four percent of the patients were referred to the C-L psychiatric service by internists. The mean age of the patients seen on this service was 31.1 (SD 13.1) years, while those of psychiatric hospital patients were 32.6 (SD14.5) (inpatients) and 33.2 (SD13.9) years (outpatients). A majority of C-L patients were female and of Indian descent, while the psychiatric hospital patients were predominantly males with more indigenous Fijian inpatients but more Indian outpatients. The most common psychiatric disorders in the C-L service were depressive episode/recurrent depressive disorder (29.9%), and neurotic, stress related and somatoform disorders (25.7%). The most common psychiatric disorders encountered in the psychiatric hospital were schizophrenia/delusional disorders (58.5% for inpatients and 52.7% for outpatients) and bipolar affective disorders (30.6% for inpatients and 27.9% for outpatients). CONCLUSION: The most likely person to utilize the C-L service in this population is a young adult female Indian who has been suffering from an internal medical condition, which became complicated by depression, resulting in the attending physician seeking psychiatric opinion. The C-L and psychiatric hospital patients are different in age, gender and racial distributions, and in patterns of psychiatric morbidity. The characteristics of this C-L service place it between what obtained in centres in developing countries and advanced ones. PMID- 12737414 TI - Attitudes of Pacific parents to circumcision of boys. AB - BACKGROUND: Circumcision for cultural reasons is routine in Pacific Island countries. In New Zealand routine circumcision for which there is no medical indication is uncommon and no longer publicly funded within the public hospital system. This has caused difficulties for the Pacific people of New Zealand. AIM: This study documents the differences in the attitudes of Pacific parents and their male children to cultural circumcision, and assesses the strength of their beliefs. METHODOLOGY: Pacific boys between the ages of 8-18 and their parents resident in Christchurch were given a questionnaire to complete and then were interviewed. The participants were obtained mainly through church organisations and after broadcast on Pacifica radio. RESULTS: One hundred and sixteen of the 123 participants felt that they had strong ties to the Pacific community. The majority (89%) of the Pacific people felt that circumcision should be performed mainly for reasons of culture and hygiene. Only a small number were aware of the possible complications that might occur with circumcision. The average age that most Pacific people were circumcised and would want their children to be circumcised is between 6 and 10 years of age. Boys were less sure than their fathers that they would get their own sons circumcised. CONCLUSION: This study has shown that circumcision is expected and surprisingly well accepted by the boys of Pacific families despite the discomfort they know the procedure causes. There is a strong cultural demand from parents for circumcision. Guidance from church leaders or sexual health lessons at schools or elsewhere may alter the cultural importance of circumcision for Pacific Island people. However, the preference for circumcision is so well entrenched into their cultural beliefs and may take years to influence. PMID- 12737415 TI - Infant feeding and feeding problems experienced by mothers of a birth cohort of Pacific infants in New Zealand. AB - OBJECTIVES: To describe the rates of exclusive breastfeeding, combination feeding, formula only feeding and infant feeding problems experienced by mothers of a birth cohort of Pacific infants in New Zealand. METHODS: The data were gathered as part of the Pacific Islands Families Study, a prospective cohort study in which 1376 mothers of a cohort of 1398 Pacific infants born in Middlemore Hospital, South Auckland, New Zealand during 2000 were interviewed at home when their infants were six weeks old. Included in the interview were questions regarding method of infant feeding and any feeding problems experienced. RESULTS: At the time of discharge from hospital, 85.5% of mothers were exclusively breastfeeding. However, six weeks later this dropped to 50%. The rates of exclusive breastfeeding (p = 0.003) and combination breast and formula feeding (p < 0.001) differed amongst the main ethnic groups. Samoan mothers had the highest exclusive rate (55.7%), and Tongan mothers had the lowest rate (42.2%). Several mothers reported experiencing common feeding problems. Reasons for combination breast and formula and formula-only feeding are discussed. CONCLUSIONS: Exclusive breastfeeding rates fell below national targets and those found in recent New Zealand studies. Most of the feeding problems reported are modifiable with education and support. IMPLICATIONS: It is suggested that the benefits of breastfeeding need promoting and increased postnatal support should be provided to increase the rates of exclusive breastfeeding amongst Pacific women. Supporting Tongan women should be a priority, given the lower exclusive breastfeeding rates observed among Tongan mothers. PMID- 12737416 TI - New Zealand resident Tongan peoples' health and illness beliefs and utilisation of the health care system. AB - This study investigated the perceptions of health and illness, and the utilisation of the health care system, over the last five years, of Tongan people who reside in New Zealand. A descriptive qualitative approach was used, with the study being implemented by networking with representatives of the Tongan community and health care workers. Twenty adult Tongan people volunteered and were given the interview questions in advance. The individual face to face semi structured interviews took approximately 30 to 45 minutes. Participants verified the transcription of their interviews. Answers were analysed by identifying themes in response to each interview question. Most participants considered health to be a state of physical, mental, social and spiritual well being. Illness beliefs were predominantly either a biological malfunction of the body, or a breakdown of the holistic state of well being. All participants had used the western orthodox medical system, with the general practitioner being consulted more than other services. A high level of satisfaction was expressed with this service. Traditional Tongan healing was utilised by about half the participants, and most were happy with its outcomes. A lesser number of participants had used alternative health care, but the levels of satisfaction with it were mixed. PMID- 12737417 TI - A tool for rapid qualitative assessment of hospital-based emergency medical services among developing health sectors. PMID- 12737418 TI - The changing face of disaster management: implications for healthcare providers in the Pacific Islands. PMID- 12737419 TI - Disaster epidemiology: prudent public health practice in the Pacific Islands. AB - Natural or technological disasters may strike a community at any time, causing death, disability, illness and material destruction. Loss of human life, healthcare costs associated with disaster-related injuries and illnesses, pain and suffering, economic loss from destruction of homes, commercial enterprises and public structures are some of the myriad ways in which the impact of the disaster may be felt by the community. Disaster managers seek to prevent, mitigate and prepare for disasters during the pre-impact phase. If and when a disaster occurs, they seek to provide timely, appropriate and effective response, relief and rehabilitation services. Disaster epidemiology arose out of a need to apply objective descriptive and analytic tools to the field of disaster management in order to improve its reievance, effectiveness, and efficiency. The table illustrates the types of studies that can be undertaken during each phase of disaster management. During the pre-impact phase vulnerability and community hazards analyses can provide useful information to public health officials responsible for prevention and mitigation of potential disasters. Epidemiologic surveys of the disaster-affected area to assess the nature and the impact of the disaster are crucial during the impact phase since the information is valuable to public health, paramedical, and medical staff in tailoring their response and relief efforts. Post-impact retrospective and prospective epidemiologic studies can assistwith the establishment of appropriate rehabilitation services. They can also serve as pre-disaster exercises since results and conclusions from these studies can help disaster managers in planning the resources needed for future disasters. Finally, there is a need for surveillance of disaster-related or emergency-related illnesses, injuries and deaths during all three phases so that baseline data is readily available for comparison during or after the disaster occurs. See Table 1. PMID- 12737420 TI - Malignant otitis externa: a review. AB - Malignant otitis externa is a rare but potentially fatal disease of the external auditory canal seen mostly among elderly, diabetic or immunocompramised patients. The causative organism is mainly Pseudomonas aeruginosa. The disease spreads rapidly, invading surrounding soft tissues, cartilage and bones causing their necrosis and even spreading to the cranial nerves. The disease can be fatal if treatment is not aggressive and timely, especially if it spreads outside the auditory canal with involvement of the cranial nerves. Treatment is mainly medical with antipseudomonal drugs like the third generation cephalosporin and the fluoroquinolones and local debridement. With aggressive treatment the mortality rate from this disease, which used to be 50% in the past has now been reduced to 10-20%. The pathophysiology of the disease, clinical presentation, diagnosis, treatment and the outcome has been discussed and reviewed. PMID- 12737421 TI - Review of drug-related legislation in the Republic of Fiji Islands. AB - The development of new drug-related legislation for Fiji commenced in 1996 when two draft Bills, namely the Pharmacy Bill and the Poisons and Therapeutics Goods Bill, which had been based on World Health Organisation (WHO) drafts, were prepared and circulated for comment. During a six-week period in 1998, the consultant analysed comments on the drafts, co-ordinated stakeholder meetings, and provided workshops for the identified major stakeholders. Implementation of the outcomes from the consultancy has been delayed by first, a democratic change in Government and the resulting establishment of new priorities, secondly by a coup with subsequent interim administration, and thirdly by return to democracy with priority reassessment by the new Government. PMID- 12737422 TI - Agricultural diversity and traditional knowledge as insurance against natural disasters. 1979. PMID- 12737423 TI - Development of prehospital emergency medical services: strategies for system assessment and planning. AB - Increasing economic capacity in the Pacific Rim has led to a greater demand for integrated pre-hospital care systems. The economic and technological growth of Taiwan, South Korea, Japan, Singapore and Hong Kong has lead to health system developments and growth of advanced healthcare and prehospital medical services. Changing economies and population distribution of many of the Southwestern countries and island regions in the South Pacific, as well as the geographical constraints necessitate a systematic and individually tailored planning process, while standardizing communication and quality of service with more will developed neighbors. While EMS systems in such a broad geographical region may take a variety of forms, each system contains some system components similar to those found in the United States and Southwest Pacific regions such as Australia and New Zealand. In evaluating EMS abroad, it is useful to compare the developing system type to one of five models: hospital based, municipal, private, volunteer, and complex. In so doing, the appropriate model system may be constructed to accommodate the demands of an evolving system. PMID- 12737424 TI - Using a community-based approach for prevention and mitigation of national health emergencies. AB - Disaster management is most effective when developed at the community level. Community based planning, prevention, mitigation, and emergency response all engage the population to make choices that are commensurate with local needs and resources. Community based disaster management also offers a force multiplying effect by increasing the number of potential participants and leaders. This discussion of key community based developmental activities is meant to provide a primer for the public health practitioner beginning the study of emergency preparedness and response. PMID- 12737425 TI - Logistical considerations for emergency response resources. AB - Resource management is a critical component of disaster preparedness and response. The type and quantity of resources and supplies needed by any particular community will be determined by several factors including the disasters affecting the community, existing resources within the community, resources available from neighboring communities, and the vulnerability assessment of a community. Ideally only needed resources should be requested and delivered. Unsolicited aid can often hamper an emergency response. The needs of a community will change during a disaster. Often the immediate need focuses on the medical sector. Issues such as hygiene, water and shelter will occur later. Disaster planning and logistical management of resources should not only consider the short-term needs of the community but also the long-term consequences of a disaster on the community. PMID- 12737427 TI - The National Pharmaceutical Stockpile Program: an overview and perspective for the Pacific Islands. AB - The National Pharmaceutical Stockpile (NSP) program was created as a national resource and is an essential response component of the Centers for Disease Control and Prevention's (CDC's) larger Bioterrorism Preparedness and Response Initiative. The role of the NPS program is to maintain a national repository of life-saving pharmaceuticals and medical supplies that can be delivered to communities in the event of a biological or chemical terrorist attack or an event involving mass casualties. The NPS is to be a re-supply and backup mechanism to state and local emergency response. Before a decision is made to deploy NPS assets, CDC will collaborate with local, state, and federal officials to determine the nature and extent of the event. Once the federal decision to deploy NPS assets is made, CDC's NPS program will arrange for delivery of assets to reach the affected area within 12 hours. PMID- 12737426 TI - Key topics for emergency health education in the Pacific. AB - This discussion of key emergency health topics is meant to provide a primer for the public health practitioner beginning the study of emergency preparedness and response. The discussion focuses on the key concepts of: general principles of disaster management; emergency operations planning; incident management systems; disaster communications' mass casualty management; hazardous material response. PMID- 12737428 TI - Pacific Region Indigenous Doctors' Congress, 2002. PMID- 12737429 TI - The University of Washington Pacific Islands Continuing Clinical Education Program (PICCEP): Guam Conference on structure and content of continuing clinical education programs in the U.S.-associated jurisdictions. PMID- 12737430 TI - Developing a public health emergency operations plan: a primer. AB - EOPs are an important component of public health emergency preparedness. EOPs are developed according to an "all-hazard approach". These EOPs include a basic plan, functional annexes, hazard-specific appendices and SOPs. PMID- 12737431 TI - The Pacific Center for Emergency Health: an anatomy of collaborative development and change--the Palau perspective. AB - Many Pacific Islands Countries (PICs) by their geographic location, isolation, and lack of resources, are at risk for both environmental and man-made disasters. Disaster management (DM) and mitigation is frustrated by the general underdevelopment of DM planning and lack of adequate emergency medical services (EMS) to deal with daily emergencies let alone large-scale emergencies and disasters. To address this, the U.S. Centers for Disease Control and Prevention (CDC) developed and implemented the Pacific Emergency Health Initiative (PEHI) to review and make recommendations regarding the current level of DM/EMS development of select PICs. As a practical next step, a collaborative demonstration project- the CDC--Palau Community College Center for Emergency Health--was established in the Republic of Palau with the purpose of providing training and technical assistance in DM/EMS development for the region. In September 2001 the Center conducted two simultaneous training programs addressing Public Health Disaster Planning (one-week) and pre-hospital First Responder Care (two-weeks). Sixty participants included public health planners, physicians, and fire and police officials from eleven PIC jurisdictions and representatives from the Secretariat of the Pacific Community, South Pacific Applied Geoscience Commission, and the Fiji School of Medicine. Eleven country and state public health disaster plans were initiated. Through CDC's PEHI additional Center training programs are planned through FY 2003. PMID- 12737432 TI - Assistance for emergency health. AB - The public health agencies of Pacific island nations have the responsibility of maintaining health during national emergencies. Assistance for completion of this task is available to the Pacific islands in the form of technical, informational, educational and humanitarian aid. Assistance for Pacific island preparedness and response may originate from local, jurisdictional, regional and international levels. The Internet also now offers many useful resources for disaster education, collaboration and aid. This article discusses mechanisms and resources that Pacific island health officials may utilize to promote emergency health within their own jurisdictions. PMID- 12737433 TI - In-country and community-based postgraduate family practice training for Micronesian physicians--the Palau AHEC: a collaborative effort. AB - The U.S. Institute of Medicine in its 1998 review of the health care systems among the U.S.-Associated Pacific Islands (USAPI) identified promotion of primary health care (PHC) and training of the regional health workforce including postgraduate training for physicians as priorities. With the support of the health leadership of the USAPI and the Republic of Palau, the John A. Burns School of Medicine (JABSOM) of the University of Hawaii captured U.S. federal Area Health Education Center (AHEC) funds to implement a postgraduate program to train Family Practitioners - physician specialists in primary care for the region. The Palau AHEC has evolved into ajoint activity of JABSOM, the University of Auckland Faculty of Medicine and Health Sciences (UAFMHS), the School of Public Health & Primary Care--Fiji School of Medicine, and Palau Community College to provide Diploma-level training in Family Practice and Community Health for Micronesian physicians. PMID- 12737434 TI - History of the Pacific Emergency Health Initiative (PEHI). AB - At the February 2000 meeting of the Pacific Island Health Officers Association (PIHOA), the Centers for Disease Control and Prevention (CDC) proposed a regional strategy for promotion of emergency environmental health in the Pacific. This CDC Pacific Emergency Health Initiative (PEHI) called for a pro-active partnership of governmental institutions, international agencies, Pacific nations, the Pacific Basin Medical Association and PIHOA. The mission of PEHI is to "strengthen the capacity for emergency health preparedness and response among Pacific island nations". This article describes the history and development of PEHI and offers insight into future efforts. PMID- 12737435 TI - Jaundice. PMID- 12737436 TI - Oesophageal intraluminal nitric oxide facilitates the acid-induced oesophago salivary reflex. AB - BACKGROUND: The present study explores some aspects of the triggering of the acid induced oesophago-salivary reflex. In addition to hydrogen ions, there are two acid-dependent molecules with messenger potential in the oesophageal lumen: CO2 and NO. The aim of this study was to clarify whether oesophageal NO and CO2 participate in the regulation of salivary neutralizing capacity in response to acid exposure. METHODS: Healthy volunteers received oesophageal acidification composed of HCl, with NO3-, or HCO3- or NO3- and HCO3- in combination. In a second series of experiments, the exposure period was divided into 2 separate 10 min events. Saliva volume and titratable buffering capacity were used to calculate alkaline secretion. RESULTS: Salivary alkaline secretion increased markedly following 20 min intraluminal exposure to HCl. The initial part of this response was 22% +/- 2.2% larger (P < 0.05) if NO3- was present. When HCO3- was added, or if NO3- and HCO3- were given simultaneously, the secretory response tended to be lower. The accumulated responses over 70 min to 2 short HCl exposures (10 min each separated by a 30 min 'rest') compared to one long one lasting 20 min were similar regardless of the presence of NO3-. CONCLUSION: The data suggest that oesophageal intraluminal NO facilitates initiation of the oesophago-salivary reflex. CO2 seems to have a negligible effect on alkaline salivation, and repeated stimulation does not influence the magnitude of the response over time. PMID- 12737437 TI - Endoscopic evaluation of the upper gastrointestinal tract is worthwhile in premenopausal women with iron-deficiency anaemia irrespective of menstrual flow. AB - BACKGROUND: In premenopausal women, iron-deficiency anaemia is common and menstrual flow is often held responsible, but it is not clear whether these women should be submitted to gastrointestinal (GI) evaluation. We aim to prospectively investigate whether premenopausal women with iron-deficiency anaemia benefit from GI evaluation regardless of menstrual flow. METHODS: The study population comprised 59 consecutive premenopausal women with iron-deficiency anaemia. Excluded were women with obvious or suspected causes of anaemia and those < or = 21 years. Heavy menstrual loss was not considered an exclusion criterion. All subjects had: complete blood count, ferritin, non-invasive testing by faecal occult blood (FOB), 13C-urea breath test (13C-UBT), anti-tissue transglutaminase antibodies (tTG) and gastrin levels. Gastroscopy with antral (n = 3), corporal (n = 3) and duodenal (n = 2) biopsies was performed in women with positive 13C-UBT or tTG titre or hypergastrinaemia. RESULTS: Heavy menstrual loss was present in 50.8%. Non-invasive tests were positive in 40/59 (67.8%): 30 had positive 13C UBT, 12 had hypergastrinaemia, 7 had positive tTG and 3 had positive FOB. Women tested positive were similar to those tested negative as far as concerned age, haemoglobin and ferritin levels and heavy menstrual flow (55% versus 42.1%). All 40 women tested positive underwent gastroscopy with biopsies. Four (10%) had bleeding-associated lesions and 34 (85%) had non-bleeding-associated lesions. As regards upper GI findings, no differences were observed between women with normal and those with heavy menstrual flow. No lower GI tract lesions were detected in the three women with positive FOB. CONCLUSIONS: Our data suggest that premenopausal women with iron-deficiency anaemia benefit from endoscopic evaluation of the upper GI tract irrespective of menstrual flow. PMID- 12737438 TI - Studies of nitric oxide generation from salivary nitrite in human gastric juice. AB - BACKGROUND: Saliva contains substantial concentrations of nitrite derived from the enterosalivary recirculation of dietary nitrate. METHODS: We have investigated factors in gastric juice influencing the fate of nitrite in swallowed saliva. When nitrite (100 microM) is added to human gastric juice pH 1.5 or pH 2.5 at 37 degrees C containing physiological concentrations of thiocyanate (1 mM) and ascorbic acid (200 microM), it is converted to nitric oxide within a few seconds. RESULTS: The reduction of nitrite to nitric oxide is slower at pH 3.5 and very little is generated at pH 4.5. The rate of nitric oxide generation at acid pH increases with increasing thiocyanate concentration. The concentration of nitric oxide generated in the above way is maintained until the ascorbic acid is depleted by the recycling of nitric oxide to nitrite. In gastric juice depleted of ascorbic acid, very little nitrite is reduced to nitric oxide at any pH. CONCLUSION: These studies indicate that in the healthy acid-secreting stomach most salivary nitrite will be reduced to nitric oxide at the gastro oesophageal junction and gastric cardia where it first encounters gastric juice. PMID- 12737439 TI - Expression and cytoprotective effect of protease-activated receptor-1 in gastric epithelial cells. AB - Thrombin is a serine protease involved in many physiological functions and its receptor. the protease-activated receptor-1 (PAR-1), has a wide tissue distribution. We hypothesized that PAR-1 is expressed in gastric epithelial cells and that thrombin can modulate defence mechanisms through PAR-1. The rat gastric epithelial cell line (RMG1) and gastric biopsy specimens from gastritis patients were used in the study. Reverse transcriptase polymerase chain reaction analysis showed that the thrombin receptors PAR-1, PAR3 and PAR-4 are expressed by RGM1 gastric epithelial cell line. Immunohistochemical and electron microspcopic studies also showed PAR-1 expression in human gastric epithelial cells. Thrombin stimulated the secretion of mucin and prostaglandin E2 (PGE2) formation in RGM1 cells in a dose-dependent manner. PAR-1 agonist also stimulated PGE2 formation. In addition, thrombin significantly increases the expression of the PGE2 receptors EP2-R and EP4-R in RGM1 cells. In conclusion, the results of the present study showed for the first time that gastric epithelial cells express thrombin receptors and that these receptors may play a protective role in the gastric mucosa. PMID- 12737440 TI - Dynamics and localization of activin A expression in rat gastric ulcers. AB - BACKGROUND: Activin A, the homodimer of the activin/inhibin betaA subunit, has been shown to participate in cutaneous wound healing. In this study we intended to determine its part in gastric ulceration. METHODS: Activin A expression was studied by immunohistochemistry and in situ hybridization in acetic-acid-induced chronic gastric ulcers in rat. The dynamics of this process were also assessed by quantitative real time RT-PCR and RNase protection assays (RPA). The effects of different doses of this cytokine on epithelial and mesenchymal cell proliferation were quantitated in vitro. RESULTS: Low amounts of activin A and its mRNA were expressed by epithelia, endothelia and fibroblasts in intact gastric tissue. Granulation tissue of gastric ulcers and gastric glands adjacent to the ulcer rim expressed markedly increased amounts of activin protein as well as activin/inhibin betaA mRNA. RPA and RT-PCR studies revealed a more than 3-fold increase in the relative abundance of this mRNA. Activin A did not affect the proliferation rate of fibroblasts and epithelial cells in vitro. CONCLUSIONS: Activin A participates in gastric ulcer healing in a similar fashion as in cutaneous wounding. Its expression on protein and mRNA level is markedly increased in ulcer base and rim. PMID- 12737441 TI - Changes in food tolerance and lifestyle after eradication of Helicobacter pylori. AB - BACKGROUND: It is known that patients with peptic ulcer disease (PUD) often have an unhealthy lifestyle that results in increased mortality because of smoking related diseases. No thorough study has been done to see what changes, if any, the patient makes to lifestyle after eradication of Helicobacter pylori. METHODS: One-hundred-and-eighty-three patients were enrolled in an open-endoscopy setting; 58% had PUD and 42% gastritis and/or duodenitis (G/D). They filled out a lifestyle questionnaire before the start of anti-Helicobacter therapy and again 1 year later. RESULTS: The prevalence of food intolerance decreased from 71% to 44% among patients with PUD (P < 0.0001) and from 76% to 63% among patients with G/D (P = 0.09). Tolerance improved for coffee, orange juice, fried foods, spicy foods and fruits. There was no significant change in smoking or alcohol consumption after eradication. Coffee and tea consumption was unchanged. Milk consumption decreased from 4.2 dL/day to 3.3 (P = 0.01). The number of meals decreased from 3.5/day to 3.4 (P = 0.005) and snacking from 1.3 snacks/day to 1.1 (P = 0.02). Consumption of fruit increased from 4.0 to 4.3 times/week (P = 0.04), but the frequency of meat, fish, vegetables, spicy foods, salty foods, sweets and cakes did not change. The time spent on each meal was unchanged. There was no change in the time spent exercising. There were few significant differences between PUD and G/D patients. CONCLUSIONS: Food was better tolerated, but there were no major changes in lifestyle after eradication of H. pylori. Patients therefore do not abuse the privilege of a more tolerant digestion by indulging in a more unhealthy lifestyle. PMID- 12737442 TI - Complementary alternative medicine in patients with inflammatory bowel disease: use and attitudes. AB - BACKGROUND: A widespread increase in the use of complementary alternative medicine (CAM) by patients with inflammatory bowel disease (IBD) has been recognized. The aim of our study was to evaluate both the extent and the determinants of CAM use by outpatients with IBD. METHODS: Outpatients of the IBD centre at the University Hospital of Berne and patients of two gastroenterology private practices in Olten (Switzerland) completed a mailed self-administrated questionnaire regarding alternative medicine. The questionnaire addressed the following topics: demographic variables; disease-related data; the use of 16 types of complementary medicine; comparison between attitudes towards alternative versus conventional medicine and out-of pocket expenses. RESULTS: Alternative medicine has been used by 47% of the patients. Diagnosis, duration and activity of disease, gender, age, previous surgery were not predictive for the use of CAM. The most commonly used CAM methods were: homeopathy, acupuncture and traditional Chinese medicine. Reasons cited for the use of CAM were: lack of satisfaction with and side effects of conventional therapy and the perceived safety of CAM. Sixty-one percent of patients noted that their IBD had improved with the use of CAM. By contrast, 16% noted a flare during CAM therapies. Forty-seven percent of patients paid more than Euro400 per year for CAM. CONCLUSIONS: Complementary medicine use is common in patients with IBD. Frequently cited reasons for the use of complementary therapies were safety of CAM; dissatisfaction with conventional therapies, including their side effects; and that CAM can be used in addition to conventional therapy. PMID- 12737443 TI - Correlation between circulating soluble ICAM-1 and prednisolone-induced amelioration of ulcerative colitis. AB - BACKGROUND: A soluble form of intercellular adhesion molecule-1 (sICAM-1) shed from endothelial cells is present in the circulation. Whether the circulating molecules represent passive turnover of surface ICAM-1 or may have some active functions in the inflammatory process is unknown. Glucocorticoids (e.g. prednisolone) are cornerstones in the treatment of acute exacerbations of ulcerative colitis (UC), and influence of the leucocyte/endothelial interaction appears to be part of their mode of action. The aim of the present study was therefore to evaluate the ICAM-1-shedding through measurements of sICAM-1 concentrations during prednisolone treatment of UC patients. METHODS: Prednisolone (40 mg) was prescribed to 15 patients with severe disease activity. At inclusion, and after 2 weeks of treatment, plasma sICAM-1 levels were measured using the ELISA technique. RESULTS: The concentrations of sICAM-1 were significantly decreased during treatment from median 256.2 (ng/ml) (interquartile range 239.7-321.0 ng/ml) to 220.4 ng/ml (196.0-276.3 ng/ml) (P < 0.01). This reduction correlated with a decrease in disease activity (r(s) = 0.8; P < 0.003). CONCLUSIONS: sICAM-1 seems to be a poor diagnostic tool, but since plasma sICAM-1 concentrations decreased during the treatment period, it might prove to be applicable as an activity marker in the individual patient. PMID- 12737444 TI - Oral budesonide significantly improves water absorption in patients with ileostomy for Crohn disease. AB - BACKGROUND: In addition to their anti-inflammatory effects, steroids influence electrolyte and water transport systems in the intestinal mucosa. This study analysed the effect of the topically acting glucocorticoid budesonide on ileostomy output in patients with Crohn disease. METHODS: Oral budesonide (3 mg/three times daily for 8 days; n = 20) was compared to placebo (n = 20) in a double-blind design using matched-pair randomization according to ileal resection length in patients without detectable inflammatory activity. Under controlled hospital conditions, absolute output volumes were measured and response was defined as a reduction in intestinal output of > 25% compared to pretreatment conditions. RESULTS: In the treatment group, we observed an absolute decrease in median intestinal output from 1,240 ml to 865 ml (30.2%), compared to 0.3% under placebo (from 950 ml to 947.5 ml). Response was documented in 60% (12/20 patients) in the treatment group compared to no response under placebo (P < 0.0001). While both treatment groups showed similar absolute median reductions (400 ml with ileal resection < or = 20 cm and 405 ml with ileal resection > 20 cm), the relative reduction (response rate) was lower in the subgroup of an ileal resection > 20 cm (36%) due to the greater increase in output secondary to the loss of ileum. CONCLUSIONS: These data support the assumption that the absorptive capacity of the intestinal mucosa for water may be improved by topically acting steroids and suggest that this occurs independently of their anti-inflammatory effect. PMID- 12737445 TI - Early cancer of the stomach arising after successful treatment of gastric MALT lymphoma in patients with autoimmune disease. AB - BACKGROUND: Extranodal marginal zone B-cell lymphoma of the mucosa associated lymphoid tissue (MALT lymphoma) arises in lymphoid tissue acquired through chronic antigenic stimulation as exemplified by Helicobacter pylori. Secondary development of gastric cancer, however, is thought to be a rare event. The detection of a signet ring cell carcinoma during follow-up endoscopy after successful therapy of MALT lymphoma in a patient with Sjogren's syndrome prompted us to analyse the frequency of subsequent gastric cancer in patients with underlying autoimmune disease (AD). METHODS: Patients with early stage MALT lymphoma and an underlying AD were evaluated for the occurrence of a secondary gastric cancer during the course of follow-up. Data analysed included the type of AD, stage of MALT lymphoma, H. pylori status, treatment for MALT lymphoma and response, follow-up, the presence of a secondary cancer, and time to development of cancer. In all patients, histologic samples were reassessed for the extent of gastritis, presence of intestinal metaplasia or focal atrophy at the time of lymphoma diagnosis. RESULTS: A total of eight patients with overt AD at the time of diagnosis of MALT lymphoma were identified. All patients were women aged between 56 and 77 years; 5 had Sjogren's syndrome, 2 had autoimmune thyroiditis (1 along with psoriasis) and 1 suffered from polymyalgia rheumatica. All patients had early stage MALT lymphoma restricted to the mucosa and submucosa at the time of diagnosis, and the presence of H. pylori was found in all cases. Two of these patients achieved complete remission (CR) of the lymphoma following H. pylori eradication, while six were judged unresponsive and underwent chemotherapy, resulting in CR in all cases. One patient died from stroke while being in CR for 2 months following chemotherapy. Two patients (25%) developed early cancer limited to the gastric mucosa while being in CR from lymphoma for 9 and 27 months, respectively, and underwent partial gastrectomy. Final staging of gastric cancer revealed pT1pN0M0 in both cases. Of the remaining 5 cases, 1 patient had a local lymphoma relapse 18 months after CR and was salvaged with radiotherapy. In the remaining 4 patients, no evidence of lymphoma recurrence or a second malignancy has been found so far by regular follow-up every 3 months for a time span between 52 and 63 months after initial diagnosis. CONCLUSION: Patients with concurrent MALT lymphoma and an underlying autoimmune condition show not only an impaired response to H. pylori eradication but might also be at increased risk for the development of gastric cancer. In view of this, such patients should be followed closely by regular endoscopies after remission of MALT lymphoma. PMID- 12737446 TI - Genetic and protein markers related to in situ growth and multiplicity in small sporadic colorectal adenomas. AB - BACKGROUND: Some early genetic events in the development of colorectal adenomas are known, but their relationship to in vivo growth characteristics is uncertain. This study compared in situ size changes and other clinicopathological variables with selected genetic and protein markers. METHODS: 56 adenomas (< or = 10 mm) from 39 patients were analysed for APC, CTNNB1 and K-ras mutations, allelic imbalance on 1p and 18q, microsatellite instability and immunohistochemical expression of HLA-DR, BAX, BCL-2 and Ki-67. For 42 of the adenomas, in situ growth was measured over 3 years. The total number of polyps in each patient was recorded. RESULTS: K-ras was mutated in 8/56 adenomas. None of the regressing adenomas revealed such mutations, compared to 20% in those that maintained or increased their size. Multivariate linear regression analysis showed that tumour growth was higher in females compared to males, and was even higher in the presence of a K-ras mutation. APC mutations were found in 37/56 adenomas. CTNNB1 mutations were found in 2/19 adenomas without APC mutation. Deletions of 1p were found in 12/56 adenomas and, seemingly, most frequent in patients with few tumours. The most frequently expressed protein was BAX (33/41), but neither this nor the other proteins showed associations with an in situ growth pattern. CONCLUSION: The multivariate linear regression model showed that patient gender and the presence of K-ras mutation had significant effects on tumour growth. The lack of the proliferative stimulus resulting from a K-ras mutation may contribute to the process of adenoma regression. PMID- 12737447 TI - Hepatitis A virus VP3 may activate serum response element associated transcription. AB - BACKGROUND: Hepatitis A virus (HAV) infection is a major public health problem worldwide. The infection does not induce any visible cytopathic effects or interfere with macromolecular synthesis in host cells. However, the hepatitis B and C viruses have recently been reported to activate intracellular signals. To clarify the effects of HAV infection on intracellular signalling, we examined the influence of 9 FLAG-tagged HAV proteins (VP2, VP3, VP1-2A, 2B, 2C, 3A, 3BC, 3C and 3D) on signal transduction pathways. METHODS: Viral protein expression vectors were co-transfected into HeLa cells with reporter plasmids controlled by a synthetic promoter containing direct repeats of the cyclic AMP response element (CRE), serum response factor (SRF), activator protein 1 (AP-1), nuclear factor kappaB (NF-kappaB) or serum response element (SRE). Cells were harvested 42 h after transfection and luciferase assays were performed. Viral protein activation twice that of the control was defined as significant. RESULTS: VP3 induced an SRE associated signal 2.2 +/- 0.3 times higher than that of control. VP3 did not activate CRE-, SRF-, AP-1- or NF-kappaB- associated signalling. The other HAV proteins tested also failed to induce these pathways. CONCLUSIONS: HAV interacts with the host signalling mechanism, and HAV VP3, different from HBX and hepatitis C core protein, may activate only SRE-associated intracellular signalling, a pathway associated with cell proliferation and differentiation. PMID- 12737448 TI - Porphyria cutanea tarda as a predictor of poor response to interferon alfa therapy in chronic hepatitis C. AB - BACKGROUND: Porphyria cutanea tarda (PCT) is sometimes associated with hepatitis C virus chronic infection. The aim of this study was to describe the effect of interferon alfa (IFN-alpha) in the treatment of patients with chronic hepatitis C and PCT. METHODS: We treated a total of 66 patients with chronic hepatitis C with IFN-alpha 2b (5 MU t.i.w.) for 12 months. Twenty-two of these patients suffered from PCT as well. These patients differed from patients without PCT in that they were men, past history of alcohol abuse and HFE gene mutations were more common and the source of infection was almost always unknown. RESULTS: Sustained virologic response was obtained in 19.7% of the 66 treated patients, 27.3% in the non-PCT group and 4.5% in the PCT group (P < 0.05). This difference could not be ascribed to the difference in sex of patients, history of alcohol abuse, HCV genotype or iron status. CONCLUSION: Multivariate logistic regression analysis revealed that PCT is independently and significantly associated with non sustained response to IFNalpha therapy. In conclusion, patients with chronic hepatitis C and PCT rarely responded to IFNalpha treatment. PMID- 12737449 TI - Secondary osteoporosis in liver transplant recipients: a longitudinal study in patients with and without cholestatic liver disease. AB - BACKGROUND: Metabolic bone disease is one of the major long-term complications in liver transplant recipients, but it remains unclear which patients are at highest risk for developing severe bone disease following transplantation. METHODS: A total of 46 consecutive, adult patients with chronic liver disease accepted for a liver transplantation waiting list were prospectively included in the study. The patients were classified into two groups: group A--chronic cholestatic liver disease (n = 28), and group B--chronic non-cholestatic liver disease (n = 18). Bone mineral density (BMD) was measured at acceptance for the waiting list and at 3, 12 and 36 months following transplantation. Markers of bone turnover (serum bone specific alkaline phosphatases (bALP), s-osteocalcin, s-1-collagen-C terminal telopeptide (1-CTP) and urine N-terminal telopeptides u-Ntx) were measured at acceptance and at 3, 6, 12, 24 and 36 months following transplantation. BMD and markers of bone turnover were compared with similar values in a matched control group of 42 healthy individuals. RESULTS: BMD decreased significantly during the early post-transplantation period (median bone loss femoral neck (FN) 3 months post-transplant 8.5%). BMD levels declined slightly from 3 to 12 months following transplantation and increased thereafter. The relative bone loss was greatest among group B patients (relative bone loss FN 3 months post-transplant: group A, 8% versus group B, 13%; P = 0.04). At 36 months, 8/17 group A and 2/9 group B patients had BMD levels that exceeded the pretransplant levels (P = 0.12). The early bone loss was positively correlated with an increase in resorption markers (s-1-CTP and u-Ntx). Group B had higher levels of both s-1-CTP and u-Ntx at 3 and 6 months post-transplant than group A patients (P = 0.03). Bone formation markers increased slowly from 6 months post transplant and onwards. Relative bone loss was positively correlated to total glucocorticoid dose during the first 3 months post-transplant. There were no differences in BMD between patients receiving tacrolimus versus those receiving cyclosporin A. CONCLUSION: Bone loss following liver transplantation is considerable in patients with both cholestatic and non-cholestatic liver disease, the first group has the poorest starting-point while the latter group has the greatest bone loss following transplantation. Bone loss is closely correlated with biochemical markers of bone resorption and total dose of glucocorticoids given post-transplant. PMID- 12737450 TI - Stenting and extracorporeal shock wave lithotripsy in chronic pancreatitis. AB - BACKGROUND: Early observational studies of endoscopic treatment and extracorporeal shock wave lithotripsy (ESWL) reported considerable or complete relief of pain in 50%-80% of patients with chronic pancreatitis. There is no consensus on the measurement of pain, making comparison of observational studies difficult, and little attention has been paid to the type and amount of analgesics used by patients before and after decompressive treatment. METHODS: We performed a retrospective study of all patients with chronic pancreatitis and large-duct disease and receiving decompressing treatment between 1 November 1994 and 31 July 1999. Primary parameters were type and amount of analgesics used. RESULTS: Forty-nine patients with chronic pancreatitis and large-duct disease received stenting of the pancreatic duct (28 patients), ESWL (6 patients) or both (15 patients). After a median follow-up of 21 months, 35% of all patients were pain-free and a further 6% were not using analgesics. Seven patients no longer needed opioids during follow-up, but five other patients needed addition of opioids to treatment. The median opioid dose among opioid-dependent patients rose slightly (9%). The median increase of weight after treatment was 1 kg per year. CONCLUSION: After decompressive treatment of patients with chronic pancreatitis and large-duct disease, we observed a small increase in weight and a small reduction in the number of opioid users. The changes may not be different from the natural course of the disease. PMID- 12737451 TI - Listeria monocytogenes and inflammatory bowel disease: detection of Listeria species in intestinal mucosal biopsies by real-time PCR. PMID- 12737452 TI - Association of protein-losing enteropathy and cryoglobulinaemia. AB - We describe a case of protein-losing enteropathy in association with cryoglobulinaemia. No underlying disorder could be definitively diagnosed, but several clinical and laboratory findings suggested an immune mediated or autoimmune disorder. We propose that the mechanism of the protein-losing enteropathy in our case was immune complex formation, complement activation and endothelial damage. PMID- 12737453 TI - The congenital duplication cyst: a rare differential diagnosis of retrosternal pain and dysphagia in a young patient. AB - Congenital cysts are malformations developing from the endoderm and mesoderm of the digestive and respiratory system in the early weeks of gestation. Unilocular or multilocular dysontogenic cysts are most commonly thoracically located adjacent to the trachea and bronchus and the development of an oesophageal duplication cyst in the oesophageal wall is extremely rare. The duplication cyst in the adult is usually asymptomatic and an incidental diagnosis. Potential symptoms include dysphagia and retrosternal pain. Next to endoscopy and computer tomography, endoscopic ultrasonography is mandatory for a distinguished and accurate preoperative evaluation. Transthoracic excision is crucial for definitive diagnosis and inhibition of complications. PMID- 12737454 TI - Chemical method to increase extreme ultraviolet microchannel-plate quantum efficiency. II. Analysis and optimization. AB - Enhancement of the extreme ultraviolet quantum detection efficiency (QDE) of microchannel plate (MCP) detectors by use of a wet chemical method is examined. It is shown that the chemical process of ion exchange, in addition to physical processes that increase surface roughness and decrease surface density, augments the secondary electron emission coefficient, which in turn increases the quantum detection efficiency of the input MCP. The method has been demonstrated with nitric acid, acetic acid, or water used as the active reactant. By monitoring and optimizing the ion-exchange process, we achieved a 2.6-4.4 increase in the MCP QDE from 1216 to 304 A, respectively, with an absolute QDE of approximately 50% at 304 A. PMID- 12737455 TI - Calculation of the efficiency of polarization-insensitive surface-stabilized ferroelectric liquid-crystal diffraction gratings. AB - A rigorous analysis is presented of the diffraction efficiency of a polarization insensitive surface-stabilized ferroelectric liquid-crystal (SSFLC) phase grating, taking full account of the internal structure of the ferroelectric liquid-crystal layer. When no field is applied, the twisted director profile in the relaxed state gives an optimum diffraction efficiency for a device thickness between the half-wave-plate and the full-wave-plate conditions. The influence of the magnitude of the spontaneous polarization and applied ac fields are investigated, and it is shown that the diffraction efficiency of a binary SSFLC phase grating can be strongly enhanced with the technique of ac stabilization. PMID- 12737456 TI - Wavelength shifts of cladding-mode resonance in corrugated long-period fiber gratings under torsion. AB - A finite deformation theory of elasticity and a theory of nonlinear photoelasticity are applied to describe the wavelength shifts of cladding-mode resonance in corrugated long-period fiber gratings under torsion. The deformation of fiber is found by use of the Murnaghan model of a solid elastic body. The quadratic photoelastic effect that is proportional to the second-order displacement gradient is investigated and compared with the classical photoelastic effect. The electromagnetic field in the twisted corrugated structure is presented as a superposition of circularly polarized modes of the etched fiber section. The wavelength shift is found to be proportional to the square of the twist angle. As predicted by our theory, a wavelength shift of the same nature has been found in a conventionally photoinduced long-period fiber grating. PMID- 12737457 TI - Wideband true-time-delay beam former that employs a tunable chirped fiber grating prism. AB - A fiber grating prism that consists of four tunable chirped-grating delay lines for wideband true-time-delay beam forming is proposed and demonstrated. The chirped gratings are produced by use of the grating bending technique in which a uniform grating is surface mounted on a simply supported beam. We obtained chirped gratings with different chirp rates by bending the uniform gratings with different beam deflections. Four linear chirped fiber gratings with identical spectral width but linearly increased grating length are fabricated. The spectra and time-delay responses of the tunable chirped gratings are measured. A chirped grating prism for wideband true-time-delay beam forming by use of four chirped gratings is constructed and tested experimentally. We obtained different time delays by tuning the wavelength of the optical carrier. The proposed true-time delay beam former with a four-element phased-array steerer is suitable for continuous beam forming at microwave frequencies up to 20 GHz. PMID- 12737458 TI - Fabrication and modeling of uniform-waist single-mode tapered optical fiber sensors. AB - We report the fabrication and modeling of single-mode tapered optical fiber sensors. The fabrication technique consist of stretching a section of fiber with an oscillating flame torch. Such a process allows controllable fabrication of lossless tapered fibers with a uniform waist. The sensor transmittance is modeled with a simple ray optics approach. In the model, all the taper parameters are taken into account. Our results indicate that sensor sensitivity can be adjusted with the taper waist diameter. As an example a gold-coated tapered fiber is theoretically and experimentally analyzed. PMID- 12737459 TI - Measuring temperature profiles in high-power optical fiber components. AB - We demonstrate a new method for measuring changes in temperature distribution caused by coupling a high-power laser beam into an optical fiber and by splicing two fibers. The measurement technique is based on interrogating a fiber Bragg grating by using low-coherence spectral interferometry. A large temperature change is found owing to coupling of a high-power laser into a multimode fiber and to splicing of two multimode fibers. Measurement of the temperature profile rather than the average temperature along the grating allows study of the cause of fiber heating. The new measurement technique enables us to monitor in real time the temperature profile in a fiber without the affecting system operation, and it might be important for developing and improving the reliability of high power fiber components. PMID- 12737460 TI - Noise-equivalent change in radiance for sampling noise in a double-sided interferogram. AB - The formula for the noise-equivalent change in radiance (NEdN) for sampling noise [Appl. Opt. 38, 139 (1999)] can work well when applied to the double-sided interferograms of radiance spectra dominated by isolated emission lines, but it does not work well when applied to broad, slowly varying radiance spectra such as a Planck blackbody curve. The modified formula for the sampling-noise NEdN works well when applied to the double-sided interferograms of both types of radiance spectra. PMID- 12737461 TI - Analysis, search, and classification for reflective ring-field projection systems. AB - Extreme ultraviolet (EUV) lithography uses reflective ring-field projection systems. Geometrical obstruction limits the possible system configurations to small domains of the parameter space. We present an analysis, a search method, and a classification of these unobstructed domains. The exhaustive search method based on paraxial analysis provides an effective means for determining all possible design forms and for finding useful starting configurations for optimization. The approach is validated through comparison with finite ray tracing. PMID- 12737462 TI - Step-zoom dual-field-of-view infrared telescope. AB - The design of a dual-field-of-view telescope for an 8-12-microm imaging waveband is described. Preliminary calculations are made to determine the first-order parameters of the narrow- and the wide-field modes. To achieve a switchable dual field-of-view system, one use an optical configuration based on the axial motion of a single lens group along the optical axis. The same lens is also used for focusing at near objects and for athermalization by small axial movement. A total of six lenses with one conic surface are used in the design, making the telescope cost effective and lightweight. The final optical design is presented, along with the aberrations curves and modulation transfer function plots, showing excellent performance in both fields of view. PMID- 12737463 TI - Emissivity modeling of metals during the growth of oxide film and comparison of the model with experimental results. AB - Emissivity modeling of metals has been developed to elucidate behavior during the growth of oxide film, and the modeling results have been compared with experimental results. To express emissivities, pseudo-optical constants of a bare metal and of an oxide film obtained by an elipsometer are substituted into the model equations. Emissivity behavior during the growth of an oxide film upon the surface of a specimen is shown in terms of spectral, directional, and polarized characteristics, and it coincides with the experimental results, both quantitatively and qualitatively. The modeling is simple and provides useful guidance for the development of emissivity-compensated radiation thermometry. PMID- 12737464 TI - Wide-spectral-range laser refractometer. AB - We present refractometric measurements made in the 266-1064-nm spectral region with a pulsed Nd:YAG laser, and the second, third, and fourth harmonics of the laser's fundamental wavelength. The critical angle is determined by the disappearance of the diffraction orders from a metal grating, forming a microcuvette with the prism's reflecting wall. A fused-silica measuring prism is used in the experiments. PMID- 12737465 TI - Shape measurement by use of liquid-crystal display fringe projection with two step phase shifting. AB - The use of an optical fringe projection method with two-step phase shifting for three-dimensional (3-D) shape measurement of small objects is described. In this method, sinusoidal linear fringes are projected onto an object's surface by a programmable liquid-crystal display (LCD) projector and a long-working-distance microscope (LWDM). The image of the fringe pattern is captured by another LWDM and a CCD camera and processed by a phase-shifting technique. Usually a minimum of three phase-shifted fringe patterns is necessary for extraction of the object shape. In this method, a new algorithm based on a two-step phase-shifting technique produces the 3-D object shape. Unlike in the conventional method, phase unwrapping is performed directly by use of an arccosine function without the need for a wrapped phase map. Hence, shape measurement can be speeded up greatly with this approach. A small coin is evaluated to demonstrate the validity of the proposed measurement method, and the experimental results are compared with those of the four-step phase-shifting method and the conventional mechanical stylus method. PMID- 12737466 TI - Interferometric characterization of phase masks. AB - We demonstrate a novel interferometric technique for highly accurate characterization of phase masks used in optical fiber grating fabrication. The principle of the measurement scheme is based on the analysis of the interference pattern formed between the first- and zero-order beams transmitted through or reflected from the grating under test. For spatial resolution of a few millimeters, our methods allow the determination of local variations of the order of 1-microm grating period with an accuracy of a few picometers. These methods are applicable to a broad class of diffractive grating structures. PMID- 12737467 TI - Angstrom-range optical path-length measurement with a high-speed scanning heterodyne optical interferometer. AB - A highly accurate method of optical path-length measurement is introduced by use of a scanning heterodyne optical interferometer with no moving parts. The instrument has demonstrated the potential to measure optical path length at angstrom resolution over continuous thickness in the micrometer range. This optical path length can be used to calculate the thickness of any material if the refractive index is known or to measure the refractive index of the material if the thickness is known. The instrument uses a single acousto-optic device in an in-line ultra-stable reflective geometry to implement rapid scanning in the microsecond domain for thickness measurements of the test medium. PMID- 12737468 TI - Frequency dispersion of electro-optical properties over a wide range by means of time-response analysis. AB - We show that a Z-transform-based time-response analysis of the electro-optical response of a crystal to a step voltage with a short rise time allows one to obtain the dispersion of the electro-optical coefficients over a wide frequency range. We describe the method employed and present the results obtained for the main electroptic coefficients (r22, r61, and rc) of a standard LiNbO3 crystal. We also show that this method is able to provide even small values of the electro optic coefficient as well as the dispersion within a wide frequency range, which is limited only by the rise time of the step voltage. PMID- 12737469 TI - Fourier-transform phase-shifting interferometry. AB - Phase-shifting interferometry is a preferred technique for high-precision surface form measurements, but the difficulty in handling the intensity distortions from multiple-surface interference has limited the general use of the technique to interferometer cavities producing strict two-beam interference. I show how the capabilities of phase-shifting interferometry can be extended to address this problem using wavelength tuning techniques. The basic theory behind the technique is reviewed and applied specifically to the measurement of parallel plates, where surfaces, optical and physical thickness, and homogeneity are simultaneously obtained. Basic system requirements are derived, common error sources are discussed, and the results of the measurements are compared with theory and alternative measurement methods. PMID- 12737470 TI - Complex refractive-index spectrum of liquid crystal in the infrared. AB - The ordinary and extraordinary complex refractive indices, n(o) - j kappa(o) and n(e) - j kappa(e), of a nematic liquid crystal were measured in the infrared region at 3.0-11.5-microm wavelength. The complex refractive indices were evaluated in terms of the angular dependence of the reflectance. Semicylindrical CsI prisms and a goniometer were used for measurement of the reflectance in a wide incident-angle range and throughout the wide infrared spectral region. Refractive indices n(o) and n(e) changed notably near the absorption wavelength. Negative birefringence, i.e., n(e) < n(o), was observed in the vicinity of 6.6 microm, where n(e) changed more than did n(o). PMID- 12737471 TI - Terahertz time-domain spectroscopy studies of the optical constants of the nematic liquid crystal 5CB. AB - The optical constants of a nematic liquid crystal, 4'-n-pentyl-4-cyanobiphenyl (5CB), in the frequency range 0.3-1.4 THz were determined by terahertz (THz) time domain spectroscopy. The real parts of the extraordinary refractive index n(e) and the ordinary refractive index n(o) of 5CB varied from 1.74 to 2.04 and from 1.59 to 1.83, respectively. Liquid-crystal 5CB exhibits a relatively small absorption loss in this frequency range. The birefringence of 5CB was found to be as large as 0.21. The experimental results indicate that liquid crystal 5CB is potentially useful for device applications in the THz frequency range. PMID- 12737473 TI - Optical quality micromachining of glass with focused laser-produced metal plasma etching in the atmosphere. AB - Recent results are reported about the optical quality surface finish obtained on glass substrates with focused laser beam produced metal plasma etching in the atmosphere. The bombardment of high-speed, high-temperature electrons from an underdense plasma on the surface of glass substrates appears to play a dominant role in this process. The effective laser fluence window for this high-quality glass machining on common microscope slides is relatively narrow. With a Corning microslide 2947, and by use of carbon steel as the plasma source, we obtained parameters between 3.5 and 4.5 J/cm2. Above the upper limit, laser-induced optical breakdown occurs in the glass material and leads to the formation of microcracks. Below the lower limit, the process was found to be ineffective. In these experiments highly defined, clean, sharp-edged, 50 x 50 micropit arrays of 15.0-microm diameter with a depth of 3.2 microm and a center-to-center separation of 18.0 microm were fabricated on Corning microslide 2947 glass substrates. PMID- 12737472 TI - Design and optical characterization of a large continuous phase plate for Laser Integration Line and Laser Megajoule facilities. AB - For development of the French Laser Integration Line and the Laser Megajoule, we describe the design and the control of a first 383 mm x 398 mm continuous phase plate prototype. Extensively used in laser fusion facilities for beam smoothing, this optical component was manufactured by deep etching onto a fused-silica substrate, which led to a phase plate engraved directly onto fused silica, with which good optical performance could be achieved. We demonstrate good agreement between the desired simulated component and the manufactured component in terms of focal spot shape. This demonstration was performed by both interferometric and photometric measurements. PMID- 12737474 TI - Micromachined, silicon filament light source for spectrophotometric microsystems. AB - A miniature broadband light source is a critical element in a spectrophotometric microsystem. The design, fabrication, and characterization of a highly stable, miniature broadband light source that comprises filaments of single-crystal silicon are presented. Electrical current versus voltage and radiant emittance spectra under constant voltage bias are measured and related to filament dimensions. A maximum stable operating temperature for these filaments is estimated to be 1200 K. Resistance drift is demonstrated to be less than 0.5% over a 10-h period of continuous operation with visible incandescence. Emittance spectra of a multifilament array, measured at three different electrical biases, are presented and shown to compare well with theoretical blackbody radiation spectra. A continuous, total radiated power of 10.7 mW was achieved with a 1 mm x 1 mm filament array with peak emittance at lambda=2.7 micrometers. PMID- 12737475 TI - Scattering from infinitely sloped surfaces by use of the Kirchhoff approximation. AB - A simple reformulation of the double-scatter Kirchhoff approximation is presented to extend the use of this calculation method to infinitely sloped surfaces. Examples are presented for square grooves and lines on perfectly conducting surfaces. The results presented show the accuracy of the method and the errors produced by not including higher-order scattering in the calculations. PMID- 12737476 TI - Application of microwave technology to design an optical multilayer bandpass filter. AB - The principles of microwave bandpass filter design are applied to design optical multilayer bandpass filters. The examples include several bandpass filters for wavelength division multiplexing. PMID- 12737477 TI - W/SiC x-ray multilayers optimized for use above 100 keV. AB - We have developed a new depth-graded multilayer system comprising W and SiC layers, suitable for use as hard x-ray reflective coatings operating in the energy range 100-200 keV. Grazing-incidence x-ray reflectance at E = 8 keV was used to characterize the interface widths, as well as the temporal and thermal stability in both periodic and depth-graded W/SiC structures, whereas synchrotron radiation was used to measure the hard x-ray reflectance of a depth-graded multilayer designed specifically for use in the range E approximately 150-170 keV. We have modeled the hard x-ray reflectance using newly derived optical constants, which we determined from reflectance versus incidence angle measurements also made using synchrotron radiation, in the range E = 120-180 keV. We describe our experimental investigation in detail compare the new W/SiC multilayers with both W/Si and W/B4C films that have been studied previously, and discuss the significance of these results with regard to the eventual development of a hard x-ray nuclear line telescope. PMID- 12737478 TI - Optical metrology for analysis of lobster-eye x-ray optics. AB - A new method that uses optical microscopy to determine the physical structure of lobster-eye x-ray optics is described. This approach offers the ability to predict x-ray performance without having to take an x-ray measurement. An overlapping series of images of the entrance and exit faces of an optic are obtained and examined by purpose-built software. A 24-parameter description of each channel is obtained from which a quantitative analysis of all the major optic defects, except surface roughness, is performed. Results for a planar lobster-eye optic are used to illustrate this technique and discuss its abilities as well as directions for future enhancements. PMID- 12737479 TI - Erbium:YAG laser lithotripsy by use of a flexible hollow waveguide with an end scaling cap. AB - An Er:YAG laser light delivery system composed of a polymer-coated silver hollow waveguide and a quartz sealing cap has been developed for calculus fragmentation. Sealing caps with various distal-end geometries were fabricated, and the focusing effects of these caps for Er:YAG laser light were measured both in air and in water. Owing to the high power capability of the quartz a beam of sealing caps, Er:YAG laser light with an output energy of 200 mJ and a repetition rate of 10 Hz was successfully transmitted in saline solution by use of the system. Calculus fragmentation experiments conducted in vitro showed that the delivery system is suitable for medical applications in lithotripsy. We also found that the cap with a focusing effect is more effective in cutting calculi. The deterioration of the sealing caps after calculus fragmentation is also discussed. PMID- 12737480 TI - Quantification of fluorophore concentration in tissue-simulating media by fluorescence measurements with a single optical fiber. AB - Quantifying fluorescent compounds in turbid media such as tissue is made difficult by the effects of multiple scattering and absorption of the excitation and emission light. The approach that we used was to measure fluorescence using a single 200-microm optical fiber as both the illumination source and the detector. Fluorescence of aluminum phthalocyanine tetrasulfonate (AlPcS4) was measured over a wide range of fluorophore concentrations and optical properties in tissue simulating phantoms. A root-mean-square accuracy of 10.6% in AlPcS4 concentration was attainable when fluorescence was measured either interstitially or at the phantom surface. The individual effects of scattering, absorption, and the scattering phase function on the fluorescence signal were also studied by experiments and Monte Carlo simulations. PMID- 12737481 TI - Temperature alterations of infrared light absorption by cartilage and cornea under free-electron laser radiation. AB - Like pure water, the water incorporated into cartilage and cornea tissue shows a pronounced dependence of the absorption coefficient on temperature. Alteration of the temperature by radiation with an IR free-electron laser was studied by use of a pulsed photothermal radiometric technique. A computation algorithm was modified to take into account the real IR absorption spectra of the tissue and the spectral sensitivity of the IR detector used. The absorption coefficients for several wavelengths within the 2.9- and 6.1-microm water absorption bands have been determined for various laser pulse energies. It is shown that the absorption coefficient for cartilage decreases at temperatures higher than 50 degrees C owing to thermal alterations of water-water and water-biopolymer interactions. PMID- 12737482 TI - Problems in assessments of amalgam electrodes for standardising or certifying the corresponding ion selective electrodes. AB - The primary method for standardizing or certifying an Mz+-ISE (Ion Selective Electrode for Mz+ where Mz+ = alkali or alkaline-earth cation) implies comparing the latter with the corresponding M-Amalgam electrode in the cell Pt/M Amalgam/Mz+ Solution/Mz+ ISE/Pt, whose potential difference is obviously independent of Mz+ concentration. Assessment of the potential of the M-Amalgam electrode requires the precise determination of the mole fraction x of the M metal in amalgam, which is customarily performed by decomposing an M-Amalgam sample in excess HCl and titrating the HCl excess with standard NaOH solution. There arises the problem of choosing the correct end-point in pH-metric titrations of strong acids with carbonate-contaminated NaOH standard solutions, which is of frequent occurrence both in research and routine laboratory practice. This topic is either overlooked or insufficiently treated in textbooks: thus the interpretation of the above experimental pH-metric titration curves is often misled and the results may be affected by significant errors. Some recent misleading suggestions are here re-analyzed critically in order to focus correct, recommended methodological schemes. PMID- 12737483 TI - Radiochemical characterisation, heavy metal determination and morphologic evolution of coastal sediments in the Ravenna area (Italy). AB - The basic aim of this work was to achieve a preliminary but integrated characterisation of an area of the Ravenna littoral coastline. The use of suitable computerised procedures has allowed the reconstruction of the morphological evolution of this area by means of historical and recent cartography. Coastal sediments were characterised by sampling 3 cores at about 2 km offshore. The following aspects were studied: 1) Mineralogical characterisation of both the total and the fine fraction of sediments by X-ray Diffraction. 2) Characteristion of sedimentary horizons by radiodating. 3)Magnetic susceptibility profiling. 4) Determination of some heavy metals (Cu, Pb, Zn, Cd) by anodic stripping voltammetry. PMID- 12737484 TI - Electroanalytical and spectroscopic characterization of poly(o-phenylenediamine) grown on highly oriented pyrolytic graphite. AB - The polymerization of ortho-phenylenediamine (o-PD) on Highly Oriented Pyrolytic Graphite (HOPG) at different pH (1,3,5,7) was investigated by electroanalytical and spectroscopic methods. Cyclic voltammetry was used both to polymerize o-PD and to study the electroactivity of the resulting poly(ortho-phenylenediamine) (PPD) film. A redox couple associated to the PPD electroactivity, deeply influenced by the pH adopted during polymerization, was recorded. A correlation between this feature and the electrochemistry shown by the oligomers of o-PD, generated in solution during the polymer synthesis, was also found. A comparison between the absorption spectra, in the visible region, of the soluble oligomers and of the PPD films was also performed, suggesting that changes in both the polymer and the oligomer structure occur and are highly related to the polymerization pH. In particular, a higher degree of conjugation is exhibited by the PPD films electrosynthesised at lower pH and this likely explains the higher conductivity as well as the higher electroactivity shown by the material obtained in these conditions. PMID- 12737486 TI - Comparison of the performances of a particle beam LC-MS interface obtained by standard and modified skimmers. AB - Stainless steel skimmers were made in the technical workshop of our Department and mounted on a commercial Particle Beam interface. Their performances were compared with the standard ones, using three different compounds: benzidine, caffeine and perylene. Library searchable EI spectra were collected with a high quality match and remarkable improvements in the sensitivity were obtained (detection limit of 20 pg for perylene and benzidine), probably due to their better tightness that allowed to widen the skimmer orifices without significantly compromising the ion source vacuum. Also calibration curves showed a good linearity, in the concentration range 10-200 ng/ml. PMID- 12737485 TI - Study of organic sulphur compounds (DMS, DMSP and CS2) in lagoon ecosystems: the case of the Venice lagoon. AB - This study of the origin and fate of dimethyl sulphide (DMS) in a particular and complex lagoon ecosystem such as that of the Venice lagoon focuses on the temporal evolutions of DMS concentrations in surface water together with those of dimethylsulphoniopropionate (DMSP), carbon disulphide (CS2), nutrients (nitrate, nitrite, ammonium, phosphate, silicate), sulphate, chlorophyll a, chlorinity, water temperature and phytoplankton (composition and density). Measurements were made from 3 March 1997 to 23 July 1998 at three stations in the central part of the Venice lagoon. The temporal trends of DMS concentration showed an absolute maximum concentration in winter (65 nmol S/l, 19/2/1998, Stn. 1; 119 nmol S/l, 19/2/1998, Stn. 2; 29 nmol S/l, 17/2/1998, Stn. 3) and two relative maxima in the spring-summer period. The spring-summer secondary maxima of DMS concentration were related to the maxima of DMSP and chlorophyll a concentrations and consequently to phytoplanktonic abundance while the winter DMS maximum showed no relation to DMSP or to chlorophyll a suggesting that the production and the fate of DMS could be different for the two periods. According to previous studies the CS2 concentration increased in the spring, achieved its maximum in summer, decreased in autumn and fell to its minimum in winter. PMID- 12737487 TI - In situ polymerization of 3-glycidoxypropyl trimethoxysilane (GLYTS) as a new tool for stone conservation. AB - The effectiveness, as stone restoration materials, of a series of polymers obtained by "in situ polymerization" of the epoxy derivative 3 glycidoxypropyltrimethoxysilane (GLYTS) with the primary amine 3 aminopropyltriethoxysilane (ATS), at various molar ratios and concentrations, has been explored through selected preliminary tests. The experiments carried on a low-porosity quartzite show that, independently from concentration and molar ratio, all of the mixtures fail to significantly affect the porosimetric and hydric properties of this stone while inducing not negligible chromatic alterations. As it concerns high porosity stones, such as Comiso calcarenite, the 2:1 15% mixture only has been selected for further experiments since preliminary tests show that it acts as an excellent barrier against water penetration either by capillarity as by total immersion while showing only negligible chromatic alterations. PMID- 12737488 TI - Optimisation of the formulation of a bubble bath by a chemometric approach market segmentation and optimisation. AB - The optimisation of the formulation of a commercial bubble bath was performed by chemometric analysis of Panel Tests results. A first Panel Test was performed to choose the best essence, among four proposed to the consumers; the best essence chosen was used in the revised commercial bubble bath. Afterwards, the effect of changing the amount of four components (the amount of primary surfactant, the essence, the hydratant and the colouring agent) of the bubble bath was studied by a fractional factorial design. The segmentation of the bubble bath market was performed by a second Panel Test, in which the consumers were requested to evaluate the samples coming from the experimental design. The results were then treated by Principal Component Analysis. The market had two segments: people preferring a product with a rich formulation and people preferring a poor product. The final target, i.e. the optimisation of the formulation for each segment, was obtained by the calculation of regression models relating the subjective evaluations given by the Panel and the compositions of the samples. The regression models allowed to identify the best formulations for the two segments ofthe market. PMID- 12737489 TI - On the complex formation equilibria between dioxouranium(VI) and sulfate ions. AB - The complexation equilibria between UO2(2+) and SO4(2-) ions have been studied at 25 degrees C in the ionic medium 3 M NaClO4 by potentiometry, by spectrophotometry and by solubility measurements of UO2(IO3)2. The potentiometric investigation was carried out with the Hg-Hg2SO4(s)-SO4(2-) half-cell and glass electrode in the sulfate concentration range 0.005 to 0.07 M. The optical absorbances in the UV-visible region and the solubility data cover the ligand concentration range 0.005 to 0.3 M. The data could be explained by assuming the complexes and equilibrium constants [Table: see text]. The constants in the infinite dilution reference state, log beta1o = 3.08 +/- 0.15 and log beta2o = 4.28 +/- 0.15, estimated by assuming the validity of the specific interaction theory, are practically coincident with literature data. PMID- 12737490 TI - On the hydrolysis of the dioxouranium(VI) ion in sulfate solutions. AB - The formation of ternary UO2(2+)-(OH-)-SO4(2-) complexes has been studied at 25 degrees C in 3 M NaClO4 ionic medium by measurements with a glass electrode. The solutions had uranium concentrations between 0.3 and 30 mM, sulfate between 20 and 200 mM, and 1.66 < or = [SO4(2-)]/[U(VI)] < or = 300. The hydrogen ion concentration ranged from 10(-3) M to incipient precipitation of basic sulfates. This occurred, depending on the metal concentration, at [H+] between 10(-4) and 10(-5.3) M. In the interpretation of the data the stabilities of binary complexes were assumed from independent sources. The data could be explained with the mixed complexes and equilibria (beta(pqr)(3sigma) refers to pUO2(2+) + qH2O + rSO4(2-) <==> (UO2)p(OH)q(SO4)r(2p-q-2r) + qH+): logbeta222 = -2.94 +/- 0.03, logbeta341 = -9.82 +/- 0.06, logbeta211 = -0.30 +/- 0.09, logbeta212 = 1.09 +/- 0.09, logbeta351 = -15.04 +/- 0.09 and logbeta462 = -14.40 +/- 0.06. The fit could be improved by including UO2OH+ with logbeta110 = -5.1 +/- 0.1. The identity of the minor species remains, however, an open question. PMID- 12737491 TI - Determination of zinc (II) in lubricant oils by stripping chronopotentiometry. AB - A method for the determination of zinc (II) in lubricant oils by stripping chronopotentiometry is described. The only necessary sample pretreatment was the extraction of zinc (II) from the corresponding alkyl derivatives by hot concentrated hydrochloric acid in a suitable extractor. The metal ions were concentrated as the corresponding metals on a glassy carbon working electrode and then stripped by a suitable oxidant. Quantitative analysis was carried out by the method of standard additions; a good linearity was obtained in the range of concentrations examined. Recoveries of 94% were obtained from a lubricant oil spiked at different levels. The detection limit was 0.02 mg g(-1) and the coefficient of variation (mean of nine determinations) was 5.2%. Results obtained on commercial lubricant oils were not significantly different from those obtained by atomic absorption spectrometry. PMID- 12737492 TI - Determination of 4-nonylphenol and 4-nonylphenol ethoxylates in river sediments by microwave assisted solvent extraction. AB - The efficiency of Microwave Assisted Solvent Extraction (MASE) for the simultaneous determination of nonylphenol (NP) and nonylphenol ethoxylates (NPE) in river sediments has been evaluated. An optimisation study was carried out in order to identify the variables having the greatest influence on the extraction efficiency. The comparison between the solvents (methanol and acetone-hexane 1:1) shows that the more polar solvent (i.e. methanol) allows a more effective extraction of NP and NPE from sediments. Analytical results show that there is not a considerable improvement by doubling extraction time, while the increase of solvent volume is significant. The comparative study with Soxhlet and pressurised liquid extraction (PLE) methods demonstrates that Microwave Assisted Solvent Extraction is a suitable alternative extraction method for the 4-NP and 4-NPE determination in river sediments, showing an accuracy and precision comparable to those obtained with PLE and better than those obtained with Soxhlet. PMID- 12737493 TI - Stability constants and thermodynamic parameters of Mn2+, Co2+, Ni2+ Cu2+, Zn2+, Cd2+, UO2(2+), Th4+, Ce3+ and Pr3+-complexes with some Schiff base hydrazones containing the pyrimidine moiety. AB - Proton-ligand dissociation and metal-ligand formation constants of 2-amino-4 chloro-6-[alpha-(phenyl)ethylidenehydrazino]pyrimidine; (AHP) and its p-chloro (ClAHP) and p-methoxy (OMeAHP) derivatives (Str.I&II) with Mn2+, Co2+ Ni2+, Cu2+, Zn2+, Cd2+, UO2(2+), Th4+, Ce3+ and Pr3+ ions have been evaluated potentiometrically in 75% (v/v) dioxane-water and 0.1 mol dm(-3) KNO3. The thermodynamic functions (deltaG, deltaH and deltaS) for the complexation of OMeAHP were evaluated and discussed. The effect of the temperature, dielectric constant of the solvents, mole fraction of dioxane and ionic strength of the medium on the stability of Pr3+-complexes show that the stability of the chelates increases by increasing both the electron repelling property of the substituents and the organic solvent content, and by decreasing the temperature, the ionic strength and the dielectric constant of the medium. PMID- 12737494 TI - A new microwave-assisted approach to the chemical analysis of mortars. PMID- 12737495 TI - The efficiency of protective treatments against damage to monuments by acid particulate. PMID- 12737496 TI - Power supply for capillary electrophoresis. PMID- 12737498 TI - The lens-coating agent and the electroretinogram. AB - PURPOSE: To investigate the effects of 0.5, 1.0 and 2.5% methylcellulose coating agents on the ERG in normals. METHODS: A total of 15 healthy volunteers underwent photopic ERG recordings in three experimental protocols comparing 0.5, 1.0 and 2.5% methylcellulose solutions. The conductivity of the solutions was measured. RESULTS: The difference between 0.5 and 1.0% solutions showed significant changes (p < 0.05) in ERG amplitudes. The 0.5% solution produced approximately 15% higher ERG amplitudes than the 1.0% solution. Comparison between 1.0 and 2.5% solutions did not show significant changes in ERG amplitudes. The conductivity was essentially the same for 0.5 and 1.0% solutions, but conductivity of the 2.5% solution was roughly half of that of the other two solutions. CONCLUSIONS: Although the 0.5% solution did yield significantly higher ERG values than the 1.0% and 2.5% solutions, the exact mechanism by which this occurs is not known. The source of these differences could arise from a combination of factors such as conductivity, viscosity, or other unknown components of the solutions. ERG laboratories should be consistent in the use of coating agents, and be aware that any change in solution might alter normative values by a modest percentage. PMID- 12737497 TI - Alzheimer's peptide and serine proteinase inhibitors in glaucoma and exfoliation syndrome. AB - The occurrence of inflammation with accompanying amyloid formation in pseudoexfoliation syndrome (PEX) resembles other inflammation-associated amyloidoses such as Alzheimer's disease (AD). To test whether the same proteins can be identified in PEX as in AD, we qualitatively analysed for Alzheimer's peptide (Abeta1-42) and the proteinase inhibitors alpha1-antichymotrypsin (ACT) and alpha-antitrypsin (AAT) in the aqueous humor of patients with and without PEX material. Ninety aqueous humor samples were collected from patients in the age group between 46 and 95 during cataract surgery. Protein profiles in samples were analysed by electrophoresis followed by Western blotting. Blots were developed using specific antibodies against Abeta1-42, AAT and ACT and peroxidase conjugated IgG as a second antibody. At least one of the analysed proteins was found in 68.8% of 90 cases studied. Abeta1-42 peptide was found in 22.2% of all cases, among them in seven cases with PEX (total n =16) and in four with glaucoma (total n = 10). ACT and AAT were detected in 17.8 and 28.9% of all cases, respectively. In addition, female patients had significantly higher frequencies of detected ACT and AAT, compared to males. Abeta1-42, ACT and AAT were also found in 17.6, 14.7 and 23.5% of the control (non-XF and non-glaucoma) samples (n = 68). Alzheimer's peptide is present in the aqueous humor of patients with PEX and glaucoma suggesting that these diseases may share common features in the biochemistry and etiologies with AD. The presence of Abeta and inflammation associated proteins in aqueous from cataract cases without detectable PEX raises the possibility that these proteins may reflect early amyloid-related changes in the eye. PMID- 12737499 TI - Passive attenuation of cortical pattern evoked potentials with increasing body weight in young male rhesus macaques. AB - The purpose of our study was to determine changes in amplitudes and implicit times of retinal and cortical pattern evoked potentials with increasing body weight in young, growing rhesus macaques (Macaca mulatta). Retinal and cortical pattern evoked potentials were recorded from 29 male rhesus macaques between 3 and 7 years of age. Thirteen animals were reexamined after 11 months. Computed tomography (CT) was performed on two animals to measure the distance between the location of the skin electrode and the surface of the striate cortex. Spearman correlation coefficients were calculated to describe the relationship between body weights and either root mean square (rms) amplitudes or implicit times. For 13 animals rms amplitudes and implicit times were compared with the Wilcoxon matched pairs signed rank test for recordings taken 11 months apart. Highly significant correlations between increases in body weights and decreases in cortical rms amplitudes were noted in 29 monkeys (p < 0.0005). No significant changes were found in the cortical rms amplitudes in thirteen monkeys over 11 months. Computed tomography showed a large increase of soft tissue thickness over the skull and striate cortex with increased body weight. The decreased amplitude in cortical evoked potentials with weight gain associated with aging can be explained by the increased distance between skin electrode and striate cortex due to soft tissue thickening (passive attenuation). PMID- 12737500 TI - The effect of body temperature on the murine electroretinogram. AB - PURPOSE: To study the effect of body temperature on the murine electroretinogram (ERG). METHODS: The corneal ERG elicited by a strobe flash from dark-adapted mice was recorded using a saline wick electrode while measuring rectal temperature continuously. The mouse was placed within a cylindrical coil of tubing through which water circulated from a temperature controlled bath. The body temperature of the mouse was changed stepwise between 30 and 37 degrees C. RESULTS: ERGs of approximately normal configuration were recorded at body temperature ranging between 30 and 37 degrees C. The maximum amplitude of the a- and b-waves varied linearly with temperature. The rate of change of b-wave amplitude was about 100 microV/degree. At 30 degrees C, maximum b-wave amplitude was about 400 microV; at 37 degrees C it was about 1000 microV. A change in body temperature produced a rapid change in ERG amplitude. CONCLUSION: The murine ERG is very sensitive to changes in temperature. In order to monitor the ERG accurately over time, continuous recording of body temperature is essential. PMID- 12737501 TI - The ERG of guinea pig (Cavis porcellus): comparison with I-type monkey and E-type rat. AB - We have been in search of an alternate species for the monkey to study the effects of drugs on the I-type photopic electroretinogram (ERG) response that is typically seen in the cone-rich retina of the primate. The guinea pig has two types of cones, one of which contains a middle-wavelength sensitive pigment otherwise found only in Old World primates. We studied the Ganzfeld electroretinogram (ERG) of the guinea pig in relation to monkey and rat ERGs to learn whether the guinea pig might be a good animal model to study the 'primate like' cone ERG. The guinea pig scotopic ERG was similar to other mammal ERGs and was not electronegative when fully dark-adapted. We saw no evidence of a negative going scotopic threshold response (STR). The guinea pig photopic ERG a-wave is larger than that of the rat but much smaller than the primate a-wave, and it lacked a phasic d-wave. PDA eliminated guinea pig photopic a-wave and caused the OFF-response to long stimuli to invert polarity, as seen in monkey but not in rat. The guinea pig overall shows a weak I-type response and may be a useful substitute for primate in some studies of the photopic ERG. PMID- 12737502 TI - The multifocal electroretinogram in X-linked juvenile retinoschisis. AB - To measure and compare the multifocal electroretinography in normal control and X linked juvenile retinoschisis, 13 cases (13 right eyes) of normal control and nine cases (17 eyes) of X-linked juvenile retinoschisis were measured with VERIS Science 4.0. Four cases (eight eyes) out of the nine retinoschisis cases were tested with Ganzfeld ERG at the same day. The results showed statistically significant difference of average response densities and latencies in six ring retinal regions between the normal control and retinoschisis. The trace array and 3-D topography of multifocal ERG showed multi-area amplitude decrease with absence or reduction of central peak amplitude in patients with retinoschisis. The P1/N1 ratio of multifocal ERG average response densities in six ring retinal regions was different from the b/a ratio of Ganzfeld ERG. The multifocal ERG and Ganzfeld ERG each had its advantage in the diagnosis of retinoschisis. PMID- 12737503 TI - Modification of the Heidelberg retinal flowmeter to record pattern and flicker induced blood flow changes. AB - PURPOSE: We tested a prototype stimulator interfaced with a commercially available scanning laser ophthalmoscope designed to measure retinal capillary perfusion (Heidelberg Retina Flowmeter (HRF)). The add-on stimulator optically superimposed the image of a monitor display on to the subject's retina coaxially with the imaging optics of the HRF. The purpose of the study was to determine if flicker and pattern stimulation presented in this manner could evoke changes in retinal perfusion that could be measured by the HRF. METHODS: The prototype stimulator projected 55 degrees visual angle circular fields of homogeneous flicker, alternating checkerboard, and multi-focal m-sequence hexagonal patterns on the retina of 10 human subjects during acquisition of images by the HRF. RESULTS: Images were successfully acquired and processed. HRF perfusion values during flicker and pattern stimulation were not significantly different from control values. CONCLUSIONS: Results of the present study and a previously published study showing a flicker-induced increase in the HRF perfusion values are contradictory. Retinal perfusion measured by the HRF were not affected by flicker and pattern stimulation delivered through the prototype device. These data are not consistent with a large flicker or pattern induced increase in retinal perfusion. The instrumental modification appears promising. However, the raster scan stimulation technique or some other aspect of stimulation or image acquisition may account for the different results in the present study and previous studies in our laboratory and in the laboratories of other investigators. PMID- 12737504 TI - Effects of betaxolol and flunarizine on visual fields and intraocular pressure in patients with migraine. AB - Fifty-one patients with migraine were divided into four groups to investigate the effects of topical betaxolol and systemic calcium channel blocker flunarizine on visual fields (VF) and intraocular pressure (IOP). The first group (Group 0) was followed with no medications, topical betaxolol (bid) was precribed to the second group (Group B), oral flunarizine (10 mg daily) was prescribed to the third group (Group F), and the last group (Group BF) was assigned for combined betaxolol and flunarizine treatment. After a mean follow-up time of 4.2 +/- 1.2 months (3-6 months), IOP measurements and VF tests were repeated. Group B and Group BF were found to be statistically different from the other groups in terms of IOP reduction and VF improvement according to mean deviation and corrected pattern standard deviation indices in the second examinations. On the other hand, Group F and Group BF differed from the other two groups considering the improvement in migrainous complaints. VF findings which are probably influenced by perfusion problems due to vasospastic mechanisms in migraineurs, improved following topical betaxolol treatment. However, systemic use of flunarizine--a calcium channel blocker--did not seem to be effective on visual fields although it had beneficial effects on migraine. PMID- 12737506 TI - Comparison of conventional ERG parameters and high-intensity A-wave analysis in a clinical setting. AB - Computational analysis of high-intensity a-waves yields direct information about the rod and cone receptor potential. However, it is not clear whether such information adds materially to the diagnostic value of the standard ERG in a routine clinical setting. We recorded both conventional ISCEV standard and computational high intensity ERG parameters from 38 patients referred to a clinical laboratory for ERG testing, and also from eight normal volunteers. The patients were grouped as: (1) macular dysfunction; (2) diffuse cone dysfunction; (3) diffuse rod-cone dysfunction. The results showed moderate variation in both conventional and computational parameters, but in general a similar pattern of normality or abnormality for both among the disease groups. There were only a few outlying subjects for which one or the other approach seemed more sensitive. We conclude that a-wave analysis is an important tool for clinical research and the study of special patients, but adding it to the standard ERG protocol does not, at our present state of knowledge, add markedly to clinical evaluations in a routine clinical setting. PMID- 12737505 TI - Retinal function in the von Hippel-Lindau disease. AB - PURPOSE: To assess the retinal function in patients with von Hippel-Lindau disease (VHL). PATIENTS: Studies were undertaken in 12 patients (17 eyes) with detected VHL gene mutation and 12 normal healthy controls (17 eyes). METHODS: Pattern ERG (PERG), standard flash electroretinogram (ERG) recordings were performed in accordance with the International Society for Clinical Electrophysiology of Vision (ISCEV) standards. RESULTS: In VHL patients, electrophysiological statistically significant changes were found. In PERG examination, increased latency of P50 was found in the total VHL group (p < 0.02) and in the VHL subgroup with retinal angiomas (p < 0.04). In ERG examination, photopic b-wave latency was increased in the total VHL group (p < 0.03) and also in the VHL subgroup without retinal angiomas (p < 0.05). In OPs, latency increase of OP2, OP3 waves and reduced amplitude of OP3 wave in the total VHL group (OP2 latency, p < 0.05; OP3 latency, p < 0.01; OP3 amplitude, p < 0.03) and in the VHL subgroup with retinal angiomas (OP2 latency, p < 0.02; OP3 latency, p < .008; OP3 amplitude, p < 0.02) were obtained. CONCLUSIONS: It can be hypothesized that dysfunction of the inner retinal layer is present in individuals with VHL disease even in patients without retinal angiomas. PMID- 12737507 TI - ERGs, cone-isolating VEPs and analytical techniques in children with cone dysfunction syndromes. AB - Photoreceptor and post-receptoral function in children with congenital and acquired cone disorders was measured by full-field electroretinogram (ERG) and transient visual evoked potentials (VEPs). Subjects were five rod monochromats (RM), five with cone dystrophy (CD), and 30 controls. Patients were diagnosed by clinical findings, ERGs, and standard color vision tests. VEP stimuli were check reversals and color grating onsets that stimulated each photoreceptor type (L-, M , or S-cones) or post-receptoral pathways (L-M, white/black). VEP signal-to-noise ratios (S/N) were calculated by Fourier analysis of VEP epochs. All RM patients showed extinguished cone ERGs. A near normal S-cone VEP was recorded from a blue cone rod monochromat without any signal from the L- or M-cone stimuli. Two other RM patients were classified as incomplete RM based on a low-level VEP signal from either L- or M-cone stimuli. CD patients had mildly to severely reduced ERGs and VEPs were abnormal to all cone-isolating stimuli. The VEP S/N ratio was not significantly correlated with the amount of rod contrast in the color stimuli. Color VEPs provide an objective assessment of macular cone function in children with cone dysfunction syndromes that is more sensitive to residual central cone function than standard full-field ERGs. VEP techniques may be useful in the early detection of cone loss in children, especially in children who do not tolerate ERG testing. PMID- 12737508 TI - Normalisation of visual evoked potentials after optic neuritis. AB - Normalisation of the visual evoked potential (VEP) in patients with optic neuritis (ON) appears to be a rare phenomenon. However, although several workers have indicated that it can happen, they have not followed up with subsequent VEP tests to confirm how long the VEP latency of the affected eye remains in the normal range. To resolve this, 18 patients with a clinical diagnosis of acute unilateral ON were followed for 5 years with repeated VEP tests to determine if the latency of the P2 wave from affected eye could return to the normal range. Furthermore, in cases where the latency returned to normal, the length of time that it remained so was also assessed. The normal range for the latency of the P2 wave was determined by measuring VEPs from a group of 18 healthy control subjects with a similar age distribution to the patients. This established an upper limit of 115.9 ms. At presentation the mean P2 latency of the affected eyes of the patients was 140 ms with a standard deviation of 16 ms. In general, the VEP latencies remained constant over the period of the investigation. However two patients demonstrated a return to normal latencies but this was only temporary. Their latencies become prolonged again within 2 years. These results provide evidence that the delayed P2 latency observed in patients with ON can return to the normal range in a small percentage of cases. However, this improvement may spontaneously deteriorateonce more as a result of further episodes of subacute demyelination. PMID- 12737509 TI - Electroretinography of short-wavelength-sensitive cones with a LED built-in electrode and its normal values. AB - To record electroretinograms (ERG) produced by short-wavelength-sensitive cone mechanisms (SWS-cone ERG), the authors used three kinds (blue, green, and red) of light-emitting diode (LED) which were built into a contact lens electrode assembly. The LEDs were used as both stimulus and background light sources. ERG was recorded using blue LED after 10 min of yellow light adaptation produced by green and red LEDs. Duration of photo-stimulation was either 2 or 100 ms. ERG recorded in normal human subjects showed two positive waves with 2 ms photo stimulation. Amplitude of the former positive wave (b1-wave) was attenuated when the luminance of yellow background increased, and the latter positive wave (b2 wave) was attenuated when the color of photo-stimulation was green or red. These findings suggest that middle-wavelength-sensitive and long-wavelength-sensitive cone mechanisms generated the former positive wave (b1-wave) and SWS-cone mechanisms generated the latter positive wave (b2-wave). Ratio of b2-wave amplitude to b1-wave-amplitude with 2 ms photo-stimulation measured on 39 normal subjects ranged from 0.5 to 2.0. It was concluded that this three-colored LED built-in electrode was useful for recording SWS-cone ERG. PMID- 12737510 TI - Central and peripheral vision loss associated with nefazodone usage. AB - A 35-year-old woman who reported persistent significant vision loss for 3 years after taking the antidepressant nefazodone was referred for electrophysiological assessment of vision. The vision changes included reduced acuities, reduced colour vision and visual field constriction in both eyes and were thought to be associated with the use of nefazodone for 6 - 8 weeks, 3 years earlier. Multifocal electroretinograms and visual evoked potentials were recorded using the Visual Evoked Response Imaging System (VERIS) to investigate the nature and site of the neural deficit. The summed retinal response showed a normal a- and b wave latency and amplitude, however, the retinal topographic mfERGs showed a severe depression of the macular response in both eyes. The cortical topographic multifocal VEP mapping also showed a central depression in the right eye compared with the left. Two-frame motion and pattern custom mfVEP were also measured to assess different forms of cortical processing and especially of motion as nefazodone has previously been associated with image persistence with moving stimuli. The responses to two frame-motion showed signs of abnormality. Thus these results suggest that the primary locus of neural damage is retinal and is likely to have resulted from neurotoxicity. Other competing hypotheses such as hysterical blindness must be ruled out. PMID- 12737511 TI - Oscillatory potentials of multifocal electroretinogram retinopathy. AB - PURPOSE: To investigate the physiological function in the posterior pole of the retina in eyes with preproliferative diabetic retinopathy using the oscillatory potentials (OPs) of the multifocal electroretinogram (mfERG). METHODS: Multifocal ERGs were recorded from 61 eyes of 61 patients with preproliferative diabetic retinopathy (pre-PDR) and 16 eyes of 16 age-similar normal control subjects. The amplitudes and implicit times of the first positive component, P1, and the OPs of the mfERGs, recorded with a slow m-sequence, from diabetics were compared with those in normal control subjects. Among the 61 eyes, the mfERGs were recorded before and after panretinal photocoagulation (PRP) in 19 eyes of 19 patients. RESULTS: The amplitudes of P1 and OPs were significantly smaller in all tested areas in the diabetics as compared with those in the normal subjects, and the degree of reduction was greater for the OPs than P1. The implicit times of P1 and OPs were significantly delayed in the diabetics in the peripheral tested regions (20-40 degrees) but not in the central area After PRP, the amplitudes of P1 were markedly reduced in all areas tested; however, the changes of the P1 implicit time were not significant. The amplitudes of the OPs were significantly reduced in the peripheral regions after PRP; however, the changes in the implicit times were not significant. CONCLUSIONS: These results indicate that retinal function in the posterior pole is markedly impaired in eyes with pre-PDR. PRP altered the mfERGs significantly, however, the reductions were limited to the amplitude. PMID- 12737513 TI - On the sensitive innervation of the ostrich's foot pads. AB - The sensitive and autonomic innervation of foot pads in the ostrich was studied employing an usual histological technique as hematoxylin-eosin or different gold chloride impregnations. The autonomic innervation is represented by isolated or grouped ganglion cells located along the course of nerve bundles. The sensitive somatic innervation is composed by free and capsulated nerve endings usually distributed in the thickness of the connective arrangement of the foot pads, in the most superficial part the first one, while the latter was generally located close to the blood vessels. The capsulated nerve endings, morphologically classified as Pacini, Pacini-like and Herbst corpuscles, show the typical structure. They are not uniformly distributed throughout the considered districts and their number are always higher in the plantar pad compared with digital pads. These corpuscles could be found isolated or assembled to organize simple flower sprays and to constitute opposito-polar corpuscles. The Authors have put forward a hypothesis on the possible functional role of the above-mentioned nerve components. PMID- 12737512 TI - Angiogenesis: basic and clinical aspects. AB - The cardiovascular system is the first functional organ system to develop in the vertebrate embryo. A widely accepted view is that blood vessels arise through two mechanisms during development, vasculogenesis and angiogenesis. New vessels in the adult arise mainly through angiogenesis, although vasculogenesis also may occur. The existence of a postnatal vasculogenesis is also supported by the evidence that both endothelial cells and endothelial precursor cells co-exist in the circulation. Angiogenesis is a biological process by which new capillaries are formed and it occurs in many physiological and pathological conditions. It is controlled by the net balance between molecules that have positive and negative regulatory activity and this concept had led to the notion of the "angiogenic switch", depending on an increased production of one or more of the positive regulators of angiogenesis. Considerable benefit can be derived in the clinical setting from manipulating angiogenesis, either positively or negatively. There is a variety of important clinical situations in which it would be desiderable to promote angiogenic processes, such as situations in which it would be desiderable to promote angiogenic processes, such as for the induction of collateral vascularization in an ischemic heart or limb. Conversely, there are pathologic conditions in which preventing angiogenic processes could be useful in the treatment of a growing tumor or a chronic inflammatory process. PMID- 12737514 TI - Study of the microcirculation of oral mucosa in healthy subjects. AB - The research has the following aims: 1: to verify the applicability of capillaroscopic investigation to oral mucosa; 2: to propose oral mucosa as an alternative to the fingernail fold for capillaroscopic investigation; 3: to describe the characteristics of the microcirculation of oral mucosa in healthy subjects. 100 healthy patients were examined. The characteristics of the micro circulation in the areas of gum mucosa and the mucosa covering of the lower lip were examined using computerised videomicroscopic techniques. For each patient we evaluated the visibility, the course, the density, the tortuosity and any images characteristic of capillary loops, besides the possible presence of microhaemorrhages, the average calibre of capillary loops and the number of capillary loops visible per square millimetre. The investigation was simple, non invasive and repeatable for each patient. An investigation of gum mucosa has revealed a course of capillary loops both parallel and perpendicular to the surface: often the tops of the capillary loops appear as regularly distributed dots or commas. Microcirculatory architecture in the area of the mucosa covering is characterised by capillary loops with a variable diameter, course and length; next to typical capillary loops with the appearance of horse stirrups, there are other loops similar to hairpins, commas and cork screws; there are also rare microhaemorrhages with the aspect of reddish stains, that could be caused by microtraumas. Visibility was very good in the area of the mucosa covering of the lower lip: mediocre in the area of gum mucosa. Our research has highlighted, that today it is possible to carry out a capillaroscopic investigation of oral mucosa in a simple and reliable way. Future research could evaluate how "normal microcirculation", that we describe in this paper, is modified during pathology PMID- 12737515 TI - Fine structure and photoperiodical seasonal changes in Pars tuberalis of hibernating bats. AB - In Pars tuberalis (PT) of pituitary gland of hibernating bats, extending cranially along the ventral face of the hypothalamic median eminence, around the hypophyseal stalk, and caudally continuing in the Pars distalis (PD), pt specific cells, follicular cells and gonadotropic cells were distinguished. Pt specific cells contain peculiar secretory granules positive to lectin WGA and negative to lectins LFA and PNA, positive to S-100 protein labeling and to PD hormones antisera. During hibernation they present a low numerical density of both secretory granules and melatonin binding sites. After light exposure, on the other hand, the latter increase in density and are associated with marked secretion synthetic activity and exocytosis. These aspects result to be more balanced in animals sacrificed during summer. These changes seem to support the hypothesis of marked annual changes even in animal species with seasonal rhythmicity of metabolisms and gonads. Follicular cells, organized in closed follicles, have slightly developed RER and Golgi apparatus during hibernation, whereas they seem to show an increased secretory activity after light exposure and during summer. In perinuclear and supranuclear cytoplasm, glycogen particles clusters (peculiar of hibernation), cilia 9+2 and multivesicular bodies were identified. Concerning FSH cells, a reduced numerical density during hibernation, the secretory granules morphological characteristics and their probable involvement in photoperiod-linked reproductive functions are investigated. PMID- 12737516 TI - Function and regulation of Aurora/Ipl1p kinase family in cell division. AB - During mitosis, the parent cell distributes its genetic materials equally into two daughter cells through chromosome segregation, a complex movements orchestrated by mitotic kinases and its effector proteins. Faithful chromosome segregation and cytokinesis ensure that each daughter cell receives a full copy of genetic materials of parent cell. Defects in these processes can lead to aneuploidy or polyploidy. Aurora/Ipl1p family, a class of conserved serine/threonine kinases, plays key roles in chromosome segregation and cytokinesis. This article highlights the function and regulation of Aurora/Ipl1p family in mitosis and provides potential links between aberrant regulation of Aurora/Ipl1p kinases and pathogenesis of human cancer. PMID- 12737517 TI - Cloning and characterization of human IC53-2, a novel CDK5 activator binding protein. AB - We have identified IC53-2, a human homologue of the rat C53 gene from a human placenta cDNA library (GeneBank Accession No.AF217982). IC53-2 can bind to the CDK5 activator p35 by in vitro association assay. IC53-2 is mapped to human chromosome 17q21.31. The IC53-2 transcript is highly expressed in kidney, liver, skeletal muscle and placenta. It is abundantly expressed in SMMC-7721, C-33A, 3AO, A431 and MCF-7 cancer cell lines by RT-PCR assay. Stable transfection of IC53-2 cDNA into the hepatocellular carcinoma SMMC-7721 cell remarkably stimulates its growth in vitro. The above results indicate that IC53-2 is a novel human gene, which may be involved in the regulation of cell proliferation. PMID- 12737518 TI - The tumor-selective over-expression of the human Hsp70 gene is attributed to the aberrant controls at both initiation and elongation levels of transcription. AB - The tumor selective over-expression of the human Hsp70 gene has been well documented in human tumors, linked to the poor prognosis, being refractory to chemo- and radio-therapies as well as the advanced stage of tumorous lesions in particular. However, both the nature and details of aberrations in the control of the Hsp70 expression in tumor remain enigmatic. By comparing various upstream segments of the Hsp70 gene for each's ability to drive the luciferase reporter genes in the context of the tumor cell lines varying in their p53 status and an immortal normal liver cell line, we demonstrated in a great detail the defects in the control mechanisms at the both initiation and elongation levels of transcription being instrumental to the tumor selective profile of its expression. Our data should not only offer new insights into our understanding of the tumor specific over-expression of the human Hsp70 gene, but also paved the way for the rational utilization of the tumor selective mechanism with the Hsp70 at the central stage for targeting the therapeutic gene expression to human tumors. PMID- 12737519 TI - Interaction with general transcription factor IIF (TFIIF) is required for the suppression of activated transcription by RPB5-mediating protein (RMP). AB - RMP was reported to regulate transcription via competing with HBx to bind the general transcription factor IIB (TFIIB) and interacting with RPB5 subunit of RNA polymerase II as a corepressor of transcription regulator. However, our present research uncovered that RMP also regulates the transcription through interaction with the general transcription factors IIF (TFIIF), which assemble in the preinitiation complex and function in both transcription initiation and elongation. With in vitro pull-down assay and Far-Western analysis, we demonstrated that RMP could bind with bacterially expressed recombinant RAP30 and RAP74 of TFIIF subunits. In the immunoprecipitation assay in COS1 cells cotransfected with FLAG-tagged RMP or its mutants, GST-fused RAP30 and RAP74 were co-immunoprecipitated with RMP in approximately equal molar ratio, which suggests that RAP30 and RAP74 interact with RMP as a TFIIF complex. Interestingly both RAP30 and RAP74 interact with the same domain (D5) of the C-terminal RMP of 118 amino-acid residuals which overlaps with its TFIIB-binding domain. Internal deletion of D5 region of RMP abolished its binding ability with both subunits of TFIIF, while D5 domain alone was sufficient to interact with TFIIF subunits. The result of luciferase assay showed that overexpression of RMP, but not the mutant RMP lacking D5 region, suppressed the transcription activated by Gal-VP16, suggesting that interaction with TFIIF is required for RMP to suppress the activated transcription. The interaction between RMP and TFIIF may be an additional passway for RMP to regulate the transcription, or alternatively TFIIF may cooperate with RPB5 and TFIIB for the corepressor function of RMP. PMID- 12737520 TI - Cloning of rat sp56, the homologue of mouse sperm ZP3 receptor-sp56. AB - Mouse sp56 is considered as one of the candidates for mouse zona pellucida 3 (mZP3) receptor. Up to date, its homologue has only been cloned from guinea pig, namely AM67. Based on the cDNA sequence of mouse sp56, we designed a pair of primer to amplify its homologue from rat testis cDNA. Using RT-PCR, two fragments of 743 bp and 938 bp were amplified. The PCR products show very high homology to mouse sp56. However, the 743 bp product completely lacks one of the seven Sushi domains of mouse sp56. Using the 743 bp product as the probe to detect the expression profile of sp56 in rat tissues, Northern blot shows that a approximately 2.0 kb mRNA expresses specifically in testis. Employed the RACE method, two full cDNA sequences of rat sp56 were obtained. A Mr approximately 42 KD band was detected in denatured and non-reducing protein sample of rat testis and sperm with anti-mouse sp56 monoclonal antibody by Western blot method. Rat sp56 was localized on rat sperm head by the indirect immunofluorescence method. Rat sp56 immunoreactivity was detected from the early pachytene spermatocytes and throughout the spermatogenesis. Its cloning will further our understanding of the mechanism of the sperm-egg recognition and binding. PMID- 12737521 TI - Isolation and functional characterization of the C-terminus of rice phosphatidylinositol 4-kinase in vitro. AB - A partial rice (Oryza sativa L.) cDNA clone, OsPI4K1c, was isolated through screening of a cDNA library constructed from tillering materials. OsPI4K1c encoded a peptide of 608 amino acids with a calculated molecular mass of 68.4 kDa. The OsPI4K1c peptide shared high homology and possessed the highly conserved domains present in most isolated cloned PI4-kinases, i.e. a lipid kinase unique (LKU) domain and a catalytic (CAT) domain. A region with similarity to pleckstrin homology (PH) domain was present in OsPI4K1c as well. Further comparison with genomic sequences in databases revealed that OsPI4K1c is located at the 3'-end of a putative rice PI 4-kinase coding gene OsPI4K1, and its coding region corresponded to the C-terminal half of OsPI4K1 protein. Twelve exons (49-562 bp in size) and 11 introns (77-974 bp in size) were identified in OsPI4K1c. The recombinant protein expressed in Escherichia coli phosphorylates phosphatidylinositol at the D4 position of the inositol ring. OsPI4K1 transcript levels were detected in a low but constitutive manner in shoot, stem, leaf, spike and root tissues and did not change upon treatment with different hormones, calcium and jasmonic acid (JA). However, treatment with salicylic acid (SA) elevated the mRNA level of the OsPI4K1 gene, which suggested the involvement of OsPI4K1 in wounding responses. PMID- 12737522 TI - Use of in vivo models to study the role of cholesterol in the etiology of Alzheimer's disease. AB - Cholesterol has been implicated in the pathogenesis of Alzheimer's disease, both through intracellular effects, and through an extracellular effect due to its physical interaction with plaque associated amyloid. Epidemiology studies have implicated high cholesterol as a risk factor for AD, and have shown that the use of cholesterol reducing agents (statins) can be protective against the disease. We, and others have shown that cholesterol levels modulate the processing of the amyloid precursor protein (APP) both in vivo and in vitro, affecting the accumulation of Abeta (Abeta) peptides which may directly impact the risk of AD. This review describes the biology of sterols, and identifies how cholesterol may exacerbate the pathogenesis of AD. Data from in vivo and in vitro studies will then be presented to describe how treatments aimed at modulating lipid levels may be efficacious in treating AD. PMID- 12737524 TI - Estrogen and neurodegeneration. AB - Although estrogen is best known for its effects on the maturation and differentiation of the primary and secondary sex organs, increasing evidence suggests that its influence extends beyond this system, and its activity in the CNS may initiate, or influence our susceptibility to neurodegenerative decline. Estrogen has been proposed to act as a neuroprotectant at several levels, and it is probable that deprivation of estrogen as a result of menopause exposes the aging or diseased brain to several insults. In addition, estrogen deprivation is likely to initiate or enhance degenerative changes caused by oxidative stress, and to reduce the brain's ability to maintain synaptic connectivity and cholinergic integrity leading to the cognitive decline seen in aged and disease afflicted individuals. PMID- 12737525 TI - Usefulness of behavioral and electrophysiological studies in transgenic models of Alzheimer's disease. AB - Over the past several years researchers have engineered many transgenic models of Alzheimer's disease. Since loss of memory is one of the major hallmarks of the disorder, the phenotypic characterization of these animals has included both behavioral tests which aim to evaluate learning abilities, and electrophysiological studies to analyze synaptic transmission and long-term potentiation, a widely studied cellular model of learning and memory. These studies are fundamental for the design of novel therapies for the treatment and/or prevention of Alzheimer's disease. PMID- 12737526 TI - Amyloid-beta immunization in Alzheimer's disease transgenic mouse models and wildtype mice. AB - Alzheimer's disease is the most prevalent form of dementia worldwide. Therapies are desperately needed to prevent and cure the disease. Mouse models of amyloid beta deposition [APP and PSAPP transgenic (tg) mice] have been useful in determining the role of amyloid-beta (A beta) in both the pathogenesis and cognitive changes in AD. In addition, they have allowed scientists to investigate potential AD therapies in living animals. Active and passive A beta immunizations have been employed successfully in APP and PSAPP tg mice to lower cerebral A beta levels and improve cognition. Optimization of immunization protocols and characterization of immune responses in wildtype mice have been reported. Based on the promising results of A beta immunization studies in mice, a clinical trial was initiated for A beta vaccination in humans with AD. Although no adverse effects were reported in the Phase I safety trials, about 5% of AD patients in the phase II clinical trial developed meningoencephalitis, ending the trial prematurely in March 2002. Studies in AD mouse models and wildtype mice may help elucidate the mechanism for these unwanted side effects and will be useful for testing newer, safer vaccines for future use in human clinical trials. PMID- 12737527 TI - Learning and memory deficits in APP transgenic mouse models of amyloid deposition. AB - Several different transgenic APP mice develop learning and memory deficits. In some cases the mice have deficits very early in life, while in other instances the mice exhibit deficits only after they have aged and amyloid deposits have accumulated. In many cases, there is a correlation in individual mice of the same age and genotype between the extent of learning and memory deficits and the amounts of deposited amyloid found in the central nervous system. While superficially this might imply that the deposited material is somehow toxic to cognition, it is likely that deposited amyloid is also an index of the overall rate of amyloid production in each mouse. Rate of production would be expected to modify not only the amounts of deposited amyloid, but also other amyloid pools, including soluble, oligomeric, conjugated (e.g. ADDLs) and intracellular. Thus, the deposited material may be an integrated reflection of total A beta production, in addition to indicating the amounts in fibrillar forms. As such, it is conceivable that other A beta pools may be more directly linked to memory deficits. Thus far, the one manipulation found to mitigate the learning and memory deficits in APP transgenic mice is immunotherapy for A beta, either using active or passive immunization against the peptide. These data together with other findings are leading to a conclusion that the fibrillar A beta deposits are not directly linked to the memory deficits in mice, and that some other A beta pool, more readily diminished by immunotherapy, is more directly linked to the mechanisms leading to poor performance in learning and memory tasks. PMID- 12737523 TI - In vivo NMR studies of neurodegenerative diseases in transgenic and rodent models. AB - In vivo magnetic resonance spectroscopy (MRS) and magnetic resonance imaging (MRI) provide unique quality to attain neurochemical, physiological, anatomical, and functional information non-invasively. These techniques have been increasingly applied to biomedical research and clinical usage in diagnosis and prognosis of diseases. The ability of MRS to detect early yet subtle changes of neurochemicals in vivo permits the use of this technology for the study of cerebral metabolism in physiological and pathological conditions. Recent advances in MR technology have further extended its use to assess the etiology and progression of neurodegeneration. This review focuses on the current technical advances and the applications of MRS and MRI in the study of neurodegenerative disease animal models including amyotrophic lateral sclerosis, Alzheimer's, Huntington's, and Parkinson's diseases. Enhanced MR measurable neurochemical parameters in vivo are described in regard to their importance in neurodegenerative disorders and their investigation into the metabolic alterations accompanying the pathogenesis of neurodegeneration. PMID- 12737528 TI - Neuroprotective mechanisms of antiparkinsonian dopamine D2-receptor subfamily agonists. AB - Numerous studies have shown that endogenous and/or environmental neurotoxins and oxidative stress may participate in the pathogenesis of Parkinson's disease (PD), but the detailed mechanisms are still unclear. While dopamine (DA) replacement therapy with L-DOPA (levodopa) improves PD symptoms, it does not inhibit the degeneration of DA neurons in the substantia nigra. Recently, bromocriptine, pramipexole and several other agonists of the dopamine D2-receptor subfamily (including D2, D3 and D4-subtypes) have been shown to have neuroprotective effects in parkinsonian models in vitro and in vivo. Their neuroprotective effects may be mediated directly and/or indirectly by antioxidant effects, mitochondrial stabilization or induction of the antiapoptotic Bcl-2 family. PMID- 12737530 TI - APP processing enzymes (secretases) as therapeutic targets: insights from the use of transgenics (Tgs) and transfected cells. AB - Secretases degrade amyloid precursor protein (APP) releasing fragments (beta peptides A beta, A beta x) that assemble to form hallmark extracellular deposits in Alzheimer's disease (AD) correlating with disease severity. As such, secretases supply targets for therapeutic intervention and form the focus of this overview. Progress in elucidating secretases and their modes of catalysis come from exploiting the use of transgenics or transfected cells. In addition to A beta x, secretases also release C-terminal fragments with putative signaling properties (amyloid intracellular domain, AICD) similar in concept to those available for conversion of the Notch-r to release the nuclear transactivator NICD. The review considers lingering questions on APP fragmentation by secretase action, ancillary proteins such as presenilins (PS1/2), nicastrin, XII, or proteases (caspases), and the influence of familial mutations (mAPP, mPS) in terms of fibrillogenesis. PMID- 12737531 TI - Organic redox-initiated polymerization process for the fabrication of hydrogels for colon-specific drug delivery. AB - Organic-redox initiated polymerization technique based on the co-initiators system comprising benzoyl peroxide and N-phenyldiethanolamine was used at ambient temperature to fabricate pH-responsive hydrogels. The effects of changes in the concentration of the co-initiators system, the ratio in which the co-initiators combined, the type of the polymerization solvent, the pH of the hydrating medium, the concentration of the cross-linking agent based on azo-bond and the pH sensitive cross-linking agent on the properties of the hydrogels were investigated. Increasing the concentration of the co-initiators system, decreasing the concentration of the two types of cross-linking agents, and replacing DMSO by ethanol as the polymerization solvent resulted in hydrogels with increased equilibrium swelling ratio and increased molecular weight between cross-links at pH 7.4. Increasing the concentration of N-phenyldiethanolamine while keeping the concentration of benzoyl peroxide constant gave hydrogels with increased equilibrium swelling ratios. The equilibrium swelling ratios of the hydrogels at pH 2.0 were not affected by the factors investigated. The polymerization technique may be suitable for the design of drug delivery systems containing thermolabile bioactive agents like peptides and proteins. PMID- 12737532 TI - Comparative evaluation of granules made with different binders by a fluidized bed method. AB - Granules were prepared using three different binders, pregelatinized starch (PGS), gelatin (GEL), and polyvinylpyrrolidone (K30) by a fluidized bed method. As a quantitative measurement of mechanical strength or abrasion resistance, granules were subjected to a friability test for certain periods of time, and friability indexes (FI) as a function of time were calculated. The data obtained were analyzed by applying standard mathematical models. According to the derived parameters of the logistic and Weibull models, which fit best to the data, mechanical strength of granules made with K30 was observed to be lower than that of the granules of PGS and GEL which have similar values of model parameters. Flow properties, consolidation, and compressibility behaviors of unfriabled (UFR) and friabled (FR) granules, which were selected based upon their Weibull time parameter, were investigated as comparative. The flow rate of granules decreased due to diminishing particle size depending on binder type and friability, but the values of angle of repose were within the acceptable limits. Regarding consolidation behavior, the change of relative density vs. the number of taps, i.e., packing rate for FR granules of GEL was slower than that of its UFR form, whereas FR granules of PGS and K30 showed faster change in relative density compared to their UFR forms. According to the parameters obtained from the Heckel equation, PGS and K30 were found to produce softer, more plastic and readily deformable granules than GEL, and the compressibility of their FR forms was not influenced negatively. PMID- 12737533 TI - Disposition of the flavonoid quercetin in rats after single intravenous and oral doses. AB - The pharmacokinetic and mean time tissue distribution parameters, after a single 50-mg/kg dose of quercetin administered as intravenous bolus, oral solution, and oral suspension, were determined using rat as an animal model. Following intravenous administration, the elimination rate constant and the elimination half-life were found to be 0.0062 min(-1) and 111 min, respectively. Examining the mean time tissue distribution parameters reflected a strong binding affinity of the drug molecules to both plasma and tissue proteins. In addition, the low permeability rate of drug molecules in the peripheral system was demonstrated. Following the oral administration of the drug, the extent of absorption was greater from solution than from suspension. Moreover, the solution showed a shorter Tmax and a higher Cmax than suspension. The absolute bioavailability for the solution was 0.275 and that for suspension was 0.162. The mean residence time (MRT) and the mean absorption time (MAT) were higher for suspension, reflecting the need for dissolving the drug in order to be absorbed. The mean (in-vivo) dissolution time (MDT(in-vivo)) was 34.5 min. Thus, an oral quercetin formulation that can readily form a drug solution in the gastrointestinal tract may enhance the absorption of the drug. PMID- 12737529 TI - Defective neurofilament transport in mouse models of amyotrophic lateral sclerosis: a review. AB - Neurofilament proteins synthesized in the cell body of neurons are assembled and transported into axons, where they influence axon radial growth, axonal transport, and nerve conduction velocities. In diseased states, neurofilaments accumulate in cell bodies and proximal axons of affected neurons, and these lesions are characteristic of many neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), spinal muscular atrophy (SMA), Charcot-Marie-Tooth disease type 2 (CMT2), and hereditary sensory motor neuropathy. Although the molecular mechanisms that contribute to these accumulations are not yet identified, transgenic mouse models are beginning to provide insight into the role of neurofilament transport in disease-related dysfunction of neurons. This review addresses axonal transport in mouse models of ALS and the special significance of neurofilament transport in this disease. PMID- 12737534 TI - Transdermal drug delivery system of haloperidol to overcome self-induced extrapyramidal syndrome. AB - Haloperidol (HAL), an antipsychotic, is associated with side effects of drug induced extrapyramidal syndrome (EPS) in conventional monotherapy. Controlled released transdermal dosage form (TDDS) of the drug was designed for maintenance therapy. Matrix-diffusion type transdermal film of HAL was designed with Eudragit NE 30D copolymer without permeation enhancer in different combinations. For the feasibility studies, all standard evaluations were performed, and their results pointed toward the suitability of TDDS. The drug release and permeation studies in Franz diffusion cell in 20% PEG-normal saline followed the Higuchi equation with optimum correlation coefficient. The neuroleptic efficacy was confirmed by maximum graded response in a rotarod apparatus. The neuroleptic-induced catatonia (EPS) in albino rats was minimum with a score of zero over a 72-hr study. The pharmacokinetic parameters in rabbit model showed a very significant prolongation of action up to 72 hr with steady-state plasma concentration (cp(ss)) of 11.58 ng/mL. Thus, the HAL-loaded TDDS improved the therapeutic profile by preventing the neuroleptic-induced EPS and might be a better alternative during its long period of psychiatric treatment over conventional dosage form. PMID- 12737535 TI - Preparation and characterization of chitosan microspheres containing doxifluridine. AB - Chitosan microspheres containing 5-fluorouracil (5-FU), tegafur (FT), and doxifluridine (DFUR) were prepared by the dry-in-oil method using silicone oil with no surfactant as a dispersion medium. For DFUR-containing chitosan microspheres (DFUR-M), reacetylation with acetic anhydride or coating using chitosan and glutaraldehyde was performed. DFUR-M, reacetylated DFUR-M, and chitosan-coated DFUR-M were investigated on in vitro drug release, and the former two microspheres were examined for in vivo degradation after subcutaneous (s.c.) implantation in mice, and in vivo plasma concentration-time profiles after s.c. implantation in rats. The present method gave fairly large microspheres purely composed of chitosan and drug because of no use of surfactant, which showed the mean particle diameters of 300-900 microm and the drug contents of 4-22% (w/w). Encapsulation efficiency of DFUR was higher than that of 5-FU and FT. DFUR-M and reacetylated DFUR-M exhibited spherical shape except chitosan-coated DFUR-M. DFUR M showed high initial rapid release, which was suppressed to some extent by reacetylation or chitosan coating. DFUR-M and reacetylated DFUR-M subcutaneously implanted were gradually degraded, and approximately half or a little more of the microspheres disappeared from the implanted site at 3 weeks postimplantation. DFUR-M and reacetylated DFUR-M implanted subcutaneously gave similar plasma concentration-time profiles of DFUR, which did not indicate prolonged release in vivo. DFUR-containing chitosan microspheres with fairly large size and good drug content could be obtained by the present preparation but remained to be improved for drug release properties. PMID- 12737536 TI - Studies on modifying the tackiness and drug release of Kollicoat EMM 30 D coatings. AB - In the search for antitack additives for Kollicoat EMM 30 D (ethyl acrylate methyl methacrylate 30% dispersion, Ph. Eur.) film coatings, various possibilities were investigated. The best results were obtained using a combination of simethicone and talc. This mixture was tested on propranolol, theophylline, and verapamil HCl blank pellets in a previously developed Kollicoat EMM 30 D basic formulation. Almost any desired drug release rate can be obtained with all three pellet formulations by varying the two pore formers hypromellose 3mPas and microcrystalline cellulose type 105. A thin application of colloidal silica onto the coated pellets additionally prevents them from sticking together during storage. PMID- 12737537 TI - Bile salt/lecithin mixed micelles optimized for the solubilization of a poorly soluble steroid molecule using statistical experimental design. AB - Bile salts and lecithin combine physiologically to form mixed micelles which aid the solubilization and absorption of dietary fats and drug molecules. In this series of experiments, we have shown how experimental design procedures aid the optimization of a formulation incorporating a bile salt, lecithin, and water with fluticasone propionate (FP) as the model poorly soluble drug. The initial inclusion of a categorical variable ruled out the use of classic response surface designs; therefore the experimental design was constructed using a d-optimal selection from a candidate set of all possible experimental combinations. A separate 2-factor central composite design was used to determine the optimum lecithin and bile salt concentrations over an extended range after the categorical variable had been eliminated. It has been demonstrated that an increase in either lecithin or cholic acid concentration produces an increase in solubility of FP, while sodium taurocholate appears to depress the solubility of FP compared with the other two bile salts. The increase in solubility associated with the increase in bile salt and lecithin is further demonstrated by a linear relationship between FP solubility and the total lipid in the formulation. The influence of molar ratio of lecithin to bile salt in the formulation is also significant. The physical properties of the mixed micellar system (solution turbidity and viscosity ranking) were used to further discriminate between formulations. The optimization showed that the dominant effect was the lecithin, which improves the solubilizing characteristics of the formulation with increasing concentration. The effect of salt concentration is less marked though slightly quadratic in nature. The overall increase in solubility demonstrated was from <1 microg/mL in water to 205 microg/mL in the optimized mixed micellar system. PMID- 12737538 TI - Preparation, optimization, characterization, and stability studies of salicylic acid liposomes. AB - Salicylic acid has been used widely in the treatment of dry skin conditions and also helps reduce acne symptoms. However, it suffers from the disadvantages of being a mild to strong irritant. Hence, its control can be achieved through encapsulation in liposomes. Liposomes were prepared by the conventional thin film hydration technique as described by Bangham et al. (J. Mol. Biol., 1965). The prepared liposomal dispersions were then characterized for entrapment efficiency, particle size by transmission electron microscopy (TEM), phase transition studies by differential scanning calorimetry (DSC), and lamellarity by nuclear magnetic resonance (NMR). The results showed the formation of bilayered liposomes in the particle size range of 0.2-0.8276 microm with a maximum entrapment efficiency of 42.6%. The liposomes stored at 4-5 degrees C demonstrated maximum stability as compared to those stored at any other temperature. PMID- 12737539 TI - An alternative derivation of the distribution of the individual bioequivalence metric. AB - A method is proposed for estimating the probability density function (PDF) of the individual bioequivalence metric. We show that under the usual assumption of independence of the various random variables, the computations are quite feasible. A program to evaluate the integrals needed for the computations has been written. Experimentation with this program shows that the pass/fail decision based on the computed PDF compares favorably with that based on the method suggested in a recent FDA Guidance (Jan. 2001). PMID- 12737540 TI - Adsorption of an amine drug onto microcrystalline cellulose and silicified microcrystalline cellulose samples. AB - The adsorption of a model amine drug (tacrine hydrochloride) from aqueous solution onto 21 microcrystalline cellulose (MCC) based samples has been investigated. The MCC source (manufacturer) affected adsorption. The adsorption appeared to be fully reversible. Adsorption was reduced by the use of high density grade MCC, high-energy milling, and silicification. Adsorption of the model drug was not affected by the particle size of the MCC. Significant variations of the adsorption characteristics between batches of certain MCC products were found. The primary mode of adsorption was by ion exchange. PMID- 12737541 TI - Permeation of naproxen from saturated solutions and commercial formulations through synthetic membranes. AB - The release of naproxen through synthetic membranes, mounted in modified Franz type diffusion cells, was evaluated, either from saturated solutions or from commercially available topical formulations containing 10% naproxen. The results obtained showed that the porous type synthetic membranes chosen (cellulose acetate and polyethersulphone) can be used for assessing product performance in quality control procedures. The formulations interacted with the solid membranes (silicone and EVA) to change their diffusional characteristics. However, transfer in the membrane, and not the formulation was rate controlling. These membranes could not therefore be used in assessing product release performance for quality control. PMID- 12737542 TI - Comparison of the reservoir competence of medium-sized mammals and Peromyscus leucopus for Anaplasma phagocytophilum in Connecticut. AB - In the northeastern United States, Anaplasma phagocytophilum, the agent of human granulocytic ehrlichiosis (HGE), is transmitted by the tick vector Ixodes scapularis. The white-footed mouse Peromyscus leucopus is a competent reservoir for this agent, but the reservoir competence of non-Peromyscus hosts of I. scapularis has not been studied. Here, we report data confirming reservoir competence of medium-sized mammals for A. phagocytophilum. Raccoons, Virginia opossums, gray squirrels, and striped skunks were live-trapped in June-August of 1998-1999 at two locations in Connecticut. Captured animals were kept for several days at the laboratory in wire-mesh cages over water to allow naturally attached ticks to drop off. Samples of blood and serum were taken from each animal prior to its release at the site of capture. Engorged ticks collected from each animal were allowed to molt. Resulting I. scapularis nymphs and adults were tested for the presence of A. phagocytophilum DNA by polymerase chain reaction, as were the blood samples from the animals. A. phagocytophilum DNA was detected in the blood of >10% of the raccoons tested. Raccoons, opossums, squirrels, and skunks produced adult I. scapularis infected with the agent of HGE. Prevalence of infection was the highest in adult ticks fed as nymphs upon raccoons (23%) and the lowest in those fed upon skunks and opossums (5-7%). The agent was present in nymphal I. scapularis fed as larvae upon raccoons and squirrels, but not in ticks fed upon skunks or opossums. We also tested the ability of I. scapularis to transmit A. phagocytophilum to laboratory-reared white-footed mice after acquiring it from medium-sized mammals. Ticks that acquired the agent from raccoons and squirrels successfully transmitted it to mice. Thus, raccoons and gray squirrels are reservoir-competent for the agent of HGE-they become naturally infected, and are capable of transmitting the infection to feeding ticks. PMID- 12737543 TI - A comparison of Campylobacter jejuni enteritis incidence rates in high- and low poultry-density counties: Michigan 1992-1999. AB - To compare the incidence of Campylobacter jejuni enteritis in high- and low poultry-density counties in Michigan between the years 1992 and 1999, an ecological study was conducted in the state of Michigan. A log-linear model was used to compare yearly, seasonal, age, and gender-specific incidence rates between county groupings. Counties with a high poultry density had a higher overall incidence of C. jejuni enteritis, particularly among children and young adults, compared with counties with low poultry density. The findings suggest that living in high-poultry-density counties is associated with higher odds for C. jejuni enteritis. This may be due to occupational exposure among poultry workers in these counties, but the findings in children suggest that indirect or environmental exposures may also play a role. Future studies should be conducted to investigate these issues. PMID- 12737544 TI - Early season crow mortality as a sentinel for West Nile virus disease in humans, northeastern United States. AB - The 1999 New York epidemic of human West Nile virus (WN) encephalitis and meningitis was preceded by a crow die-off also caused by WN infection. As one component of the subsequently developed national surveillance system, crow mortality data were collected to detect WN activity before humans might become infected. However, predicting areas at risk for human WN disease likely requires assessment of multiple factors, including the intensity and timing of crow epizootics. To identify early season measures of WN activity in crows associated with subsequent WN disease in humans, county-level crow mortality data from seven northeastern states were analyzed. A predictive model was developed based on analysis of 2000 surveillance data and then assessed for 2001. To characterize the intensity of early season WN activity in crows, 15 variables were constructed from surveillance data of 52 counties that tested at least four crows during the early season (defined as June 17-July 28, 2000). County values for each variable were dichotomized at the 75th percentile into "high" and "low" activity. Multivariate analysis indicated that "high" early season activity of two variables-density of reported dead crow sightings (reported dead crows/area) and [(WN-infected crows/tested crows) x (human population)]--were associated with report of at least one human WN disease case (for each variable: adjusted odds ratio, 6.9; 95% confidence interval, 1.2-40.6). An assessment of this model using 2001 surveillance data from 61 counties yielded similar findings. With emphasis on early season WN activity, crow surveillance may allow timely targeting of interventions to protect the public health. PMID- 12737545 TI - Spatial analysis of West Nile virus: rapid risk assessment of an introduced vector-borne zoonosis. AB - The distribution of human risk for West Nile virus was determined by spatial analysis of the initial case distribution for the New York City area in 1999 using remote sensing and geographic information system technologies. Cluster analysis revealed the presence of a statistically significant grouping of cases, which also indicates the area of probable virus introduction. Within the cluster, habitat suitability for potentially infective adult mosquitoes was measured by the amount of vegetation cover using satellite imagery. Logistic regression analysis revealed satellite-derived vegetation abundance to be significantly and positively associated with the presence of human cases. The logistic model was used to estimate the spatial distribution of human risk for West Nile virus throughout New York City. Accuracy of the resulting risk map was cross-validated using virus-positive mosquito sample sites. These new epidemiological methods aid in rapid entry point identification and spatial prediction of human risk of infection for introduced vector-borne pathogens. PMID- 12737546 TI - Ultrastructural study of the infection process of Rickettsia conorii in the salivary glands of the vector tick Rhipicephalus sanguineus. AB - This work was designed to study the infection process of Rickettsia conorii in the salivary glands of experimentally infected Rhipicephalus sanguineus ticks. One hundred six uninfected engorged nymphs were intracelomically inoculated with approximately 2 x 10(3) plaque-forming units of a rickettsial suspension. After the molt, unfed and fed adults were dissected, and the salivary glands were extracted and processed for transmission electron microscopy observation. Three different uninfected control groups were used for (1) evaluating the impact of the inoculation procedure, (2) establishing the feeding period of infected ticks, and (3) ultrastructural characterization of the salivary glands. Overall, 75.5% (80 of 106) of the nymphs inoculated with rickettsiae died during the molt or soon after hatching into adult instars; 50% (12 of 24) of the remaining infected adults showed severe malformations compromising their viability. In apparently healthy specimens, time of engorgement was longer. The contrast with the negative control groups was statistically significant, suggesting that R. conorii exerts a strong negative effect on the vector ticks. The ultrastructural study showed that in the salivary glands of infected ticks, rickettsial growth occurs preferentially in central, peripheral, and interstitial acini cells. PMID- 12737547 TI - Q fever in humans and animals in the United States. AB - Coxiella burnetii, the etiologic agent of Q fever, is a worldwide zoonotic pathogen. Although Q fever is present in the United States, little is known about its current incidence or geographic distribution in either humans or animals. Published reports of national disease surveillance, individual cases, outbreak investigations, and serologic surveys were reviewed to better characterize Q fever epidemiology in the United States. In national disease surveillance reports for 1948-1986, 1,396 human cases were reported from almost every state. Among published individual case reports and outbreak investigations, occupational exposures (research facilities, farm environments, slaughterhouses) were commonly reported, and sheep were most frequently implicated as a possible source of infection. In studies conducted on specific groups, livestock handlers had a significantly higher prevalence of antibodies to C. burnetii than did persons with no known risk. Animal studies showed wide variation in seroprevalence, with goats having a significantly higher average seroprevalence (41.6%) than sheep (16.5%) or cattle (3.4%). Evidence of antibody to C. burnetii was reported among various wild-animal species, including coyotes, foxes, rodents, skunks, raccoons, rabbits, deer, and birds. This literature review suggests that C. burnetii is enzootic among ruminants and wild animals throughout much of the United States and that there is widespread human exposure to this pathogen. Sheep and goats appear to be a more important risk for human infection in the United States than cattle or wild animals, and research studies examining the natural history and transmission risk of Q fever in sheep and goats in this country should be encouraged. PMID- 12737549 TI - Moellerella wisconsensis isolated from the oral cavity of a wild raccoon (Procyon lotor). AB - This report describes the isolation of Moellerella wisconsensis from the oral secretions of a wild raccoon in Northern California. Human enteric disease has previously been associated with this organism. This represents the first isolation of this rare enterobacterial species from a non-captive animal and only the third from a non-human source. PMID- 12737548 TI - Vector competence of Culex tarsalis from Orange County, California, for West Nile virus. AB - To evaluate the vector competence of Culex tarsalis Coquillett for West Nile virus (WN), females reared from larvae collected in Huntington Beach, Orange County, CA, were fed on 2-3-day-old chickens previously inoculated with a New York strain (Crow 397-99) of WN. The Cx. tarsalis mosquitoes were efficient laboratory vectors of WN, with estimated transmission rates of 81% and 91% for mosquitoes that ingested 10(6.5) or 10(7.3) plaque-forming units of WN/mL of blood, respectively. Based on efficiency of viral transmission and the role of this species in the transmission of the closely related St. Louis encephalitis virus, Cx. tarsalis should be considered a potentially important vector of WN in the western United States. PMID- 12737550 TI - A suite of novel allenes from Australian melolonthine scarab beetles. Structure, synthesis, and stereochemistry. AB - A suite of allenic hydrocarbons, previously unknown as a molecular class from insects, has been characterized from several Australian melolonthine scarab beetles. The allenes are represented by the formula CH(3)(CH(2))(n)()CH=*=CH(CH(2))(7)CH(3) with n being 11-15, 17 and 19, and thus, all have Delta(9,10)-unsaturation. These structures have been confirmed by syntheses and comparisons of spectral and chromatographic properties with those of the natural components. The enantiomers of (+/-)-Delta(9.10)-tricosadiene and Delta(9,10)-pentacosadiene were separable on a modified beta-cyclodextrin column (gas chromatography), and the natural Delta(9,10)-tricosadiene (n = 11) and Delta(9,10)-pentacosadiene (n = 13) were shown to be of >85% ee. Syntheses of nonracemic allenes of known predominating chirality were acquired using both organotin chemistry and sulfonylhydrazine intermediates, and comparisons then demonstrated that the natural allenes were predominantly (R)-configured. PMID- 12737551 TI - Oxidatively truncated docosahexaenoate phospholipids: total synthesis, generation, and Peptide adduction chemistry. AB - The recent immunological detection of extraordinarily high levels of carboxyethylpyrrole (CEP) modifications of proteins from the retinas of individuals with age-related macular degeneration provided presumptive evidence for the involvement of docosahexaenoate-derived oxidatively truncated phospholipids in retinal pathology. To facilitate the in vivo detection and characterization of the chemistry and biological activities of these postulated naturally occurring molecules, a family of oxidatively truncated phospholipids was prepared by total syntheses. Their formation in oxidation reactions of a docosahexaenoate ester of 2-lysophosphatidylcholine (DHA-PC) was also demonstrated. Free radical-induced oxidative cleavage of DHA-PC promoted by myeloperoxidase or copper ions generates similar mixtures of these phospholipids. The most abundant products were 1-palmitoyl-2-succinoyl-sn-glycero-3 phosphatidylcholine (4.7%) and 2-(6-carboxy-4-oxohex-5-enoyl)-1-palmitoyl-sn glycero-3-phosphatidylcholine (1.7%). Both of these oxidatively truncated phospholipids are homologues of biologically active arachidonate-derived phospholipids. A minor product from DHA-PC, 2-(4-hydroxy-7-oxohept-5-enoyl)-1 palmitoyl-sn-glycero-3-phosphatidylcholine (0.4% yield), reacted with the epsilon amino group of a peptide lysyl residue to produce a CEP derivative in 0.7% yield. These observations support the previous conclusion, based on immunological evidence, that CEPs are generated by the reaction of an oxidatively truncated phospholipid with proteins in the retina and further indicate that CEP protein modifications probably represent only a tiny fraction of the products generated upon oxidative damage of DHA-PC in photoreceptor disk membranes. PMID- 12737552 TI - Thermolysis of vic-dihydroxybacteriochlorins: effect of the nature of substrates in directing the formation of chlorin-chlorin dimers with fixed and flexible orientations and their preliminary in vitro photosensitizing efficacy. AB - The thermolysis products obtained by refluxing a series of vic-dihydroxychlorins in o-dichlorobenzene are characterized. Depending on the nature of substrates, this methodology provides an access for novel carbon-carbon linked chlorin chlorin dimers and chlorin-porphyrin dimers with fixed and flexible orientations. The configuration of the linkers in the symmetrical and unsymmetrical dimers was confirmed by extensive NMR (COSY, ROESY) and molecular modeling studies. The molecular modeling studies of the energy-optimized dimers with flexible orientation confirmed that one of the chlorin units of the dimeric structure is tilted toward the opposite ring as evident by the shielding effect in the resonances of some of the protons in the (1)H NMR spectroscopy. Among the dimers with fixed orientation, compared to the free-base analogues, the related mono- and di-Zn(II) complexes produced a decreased fluorescence intensity, suggesting a possibility of the faster energy transfer via intersystem crossing (ISC) in the metalated derivatives than the corresponding free-base analogues to produce the corresponding excited triplet states. The photosensitizing efficacy of the monomers and the related dimers was also compared in radiation-induced fibrosarcoma (RIF) tumor cells at variable drug/light doses. In preliminary screening, compared to monomers, the corresponding carbon-carbon linked dimers produced enhanced photosensitizing efficacy. PMID- 12737553 TI - A first theoretical study on the origin of the metal-mediated regioselective opening of 2,3-epoxy alcohols. AB - Hybrid density functional theory (B3LYP) method allows the study of the role of metal ions (Li(+)) in the regioselective opening of 2,3-epoxy alcohols with lithium halides (Cl, Br, I) to the corresponding halohydrins. The theoretical results largely confirm the experimental results, especially with regards to the regioselectivity observed in the opening of the oxirane ring. The C3 halogen attack is always preferred to the C2 attack, with a significant difference in the TS structures with the reaction pathway under kinetic control. The central role of the lithium cation, linked to the oxygen atoms of the epoxy alcohol, is well documented in the reaction mechanisms presented. The energy differences between the two structures of the proposed TS appear to be ascribed to the different contribution of the X-H-O hydrogen bond between the hydroxyl groups of the reactant and the incoming halogen nucleophile. PMID- 12737554 TI - Increased enantioselectivity in the addition of diethylzinc to benzaldehyde by the use of chiral ligands containing the alpha-phenylethylamino group in combination with achiral ligands. AB - Chiral ligands (S,S)-1, (S,S)-2, (S,S)-3, (S)-4, (S)-5, (S,S)-6, (S,S)-7, and (S,S)-8 turned out to be effective promoters in the enantioselective addition of diethylzinc to benzaldehyde. Interestingly, diamine (S,S)-3 and amino alcohols (S)-5 and (S,S)-7 induce the preferential formation of carbinol (R)-10 (unlike stereoinduction) whereas amido analogues (S,S)-2, (S)-4, and (S,S)-6 favor (S)-10 (like stereoinduction). Molecular modeling at the semiempirical PM3 level provided a reasonable interpretation based on conformational effects in the corresponding transition structures. Combinations of chiral ligands 1-8 with an achiral, flexible ligand (9) gave rise to an activated catalytic system that resulted in faster and higher yielding reactions. Furthermore, substantial increases in the observed enantiomeric excesses of product 10 confirmed the relevant role of achiral bis(sulfonamide) 9 as activator and "chiral environment amplifier". PMID- 12737555 TI - Influence of carbocation stability in the gas phase on solvolytic reactivity: beyond bridgehead derivatives. AB - The intrinsic gas-phase stability of bicyclic secondary carbocations has been determined by Dissociative Proton Attachment of chlorides and alcohols, respectively. From these data, Gibbs free energies for hydride transfer relative to 1-adamantyl (Delta(r)G degrees (8,exp)) are derived after application of appropriate leaving group corrections, and good agreement with theoretical values, (Delta(r)G degrees (8,comp)), calculated at the G2(MP2) or MP2/6 311G(d,p) level, is reached (Table 1). The relative rate constants for solvolysis (log(k/k(0))) of the bicyclic secondary derivatives correlate with the stabilities of the respective carbocations in the same manner as tertiary bridgehead derivatives, but simple monoderivatives and acyclic derivatives solvolyze faster than predicted on the grounds of the ion stabilities. The corresponding stabilities of cyclopropyl- and benzyl-substituted carbocations have been obtained by a combination of experimental and computational data available in the literature with computational methods. Correlation of the rate constants for solvolysis vs ion stabilities for these compounds reveals a trend similar to that observed for bridgehead derivatives, but with much more scatter, which is attributed to nucleophilic solvent participation and/or nucleophilic solvation. PMID- 12737556 TI - Thermal behavior of [2.1.1]propellane: a DFT/ab initio study. AB - Density functional and ab initio molecular orbital calculations have been used to search for the low energy path of the thermal isomerization of [2.1.1]propellane 1. Three reaction modes were considered: ring opening of the bicyclo[1.1.0]butane unit in 1 to give 1,2-dimethylenecyclobutane 21, opening of the four-membered ring of 1 to afford 1,3-dimethylenecyclobutane 20, and breaking of the [2.1.1]propellane central bond and one of the bicyclo[1.1.0]butane side bonds to form carbene 17. At the CAS(12,12)PT2N/6-31G(d) level of theory, the activation barrier of the latter route was lowest in energy. Further investigation of this process at the QCISD(T)/6-311G(d,p)//QCISD/6-31G(d) and B3PW91/6-311G(d,p)// B3PW91/6-311G(d,p) level of theory indicated that the barrier of isomerization of 1 --> 17 amounts to 29 kcal/mol and that 17 is stabilized by hydrogen migration to give dienes 18 and 19. PMID- 12737557 TI - Bis(pyridine)-based bromonium ions. Molecular structures of bis(2,4,6 collidine)bromonium perchlorate and bis(pyridine)bromonium triflate and the mechanism of the reactions of 1,2-bis(2'-pyridylethynyl)benzenebrominum triflate and bis(pyridine)bromonium triflate with acceptor olefins. AB - 1,2-Bis(2'-pyridylethynyl)benzenebromonium triflate (4) and bis(pyridine)bromonium triflate (5) have been prepared and the mechanism of their reaction with various acceptors including eight alkenes of various structure, collidine, and Br(-) are reported. The reaction of 4 with neutral acceptors is second-order overall and involves a preequilibrium dissociation of the bidentate bound Br(+) to form an unstable monodentate open form (4-op), which reacts with all neutral acceptors at or near the diffusion limit. Br(-) reacts with 4 by a different mechanism involving a direct nucleophilic attack on the Br(+). The reaction of 5 with acceptors proceeds by a dissociative mechanism to reversibly form an unstable intermediate (pyr-Br(+)), which reacts with 4-penten-1-ol, 4 pentenoic acid adamantylidineadamantane and cyclohexene with nearly the same selectivity. The crystal and molecular structures of bis(2,4,6 collidine)bromonium perchlorate (2-ClO(4)) and 5 were determined by X-ray crystallography. PMID- 12737558 TI - Novel acid-catalyzed rearrangement of methanofullerenes bearing an alpha-ylidic ester to cyclopentanofullerenes: a vinyl cyclopropane-type ring expansion. AB - A novel acid-catalyzed ring expansion of methanofullerenes bearing an alpha ylidic ester has been investigated. Treatment of dialkyl acetylenedicarboxylate and tricycloalkylphosphine with C(60) led to the isolation of a methanofullerene ylide after passing the reaction mixtures through a basic alumina column. If the reaction mixture was passed through a silica gel column, a cyclopentanofullerene was isolated instead. These new cyclopentanofullerenes consisted of a fused cyclopentanone ring bearing an alpha-hydroxy ester and a phosphonium ylide and were confirmed by their NMR, mass, and X-ray diffraction data. The cyclopentanofullerenes were formed by the ring expansion of the corresponding methanofullerenes in the presence of silica gel. The ring expansion also proceeded by treating methanofullerene with acetic acid in chloroform. On the other hand, the methanofullerenes from RO(2)CCCCO(2)R, PAr(3), and C(60) were stable in silica gel. However, upon heating with acetic acid at 50 degrees C, they underwent ring expansion and dephosphination to give cyclopentenofullerenes. An implicit vinyl cyclopropane ring expansion mechanism was proposed to account for this novel acid-catalyzed rearrangement. PMID- 12737559 TI - Synthesis and structure of novel 1lambda4,2,6-thiadiazines. AB - S,S-Disubstituted sulfodiimines 9 are known to be versatile building blocks for the synthesis of various sulfur-nitrogen-containing heterocycles. Addition condensation reactions of 9 with three different activated carbonylsubstrates 14 16 lead to several new 1lambda(6),2,6-thiadiazine-3-ones 17-20. S-Debenzylation via a novel synthetic approach utilizing different electrophiles such as TMSCl, Alk(3)O(+)BF(4)(-), Tos(2)O, and Mes(2)O gives rise to a variety of so far hardly known 1lambda(4),2,6-thiadiazine-3-ones 25 and 26. Structure elucidation reveals NH/CH tautomers in solution as well as a tetrahedral asymmetric sulfur attached to the conjugated planar N/C skeleton. The structural features of the 1lambda(4),2,6-thiadiazines 25 and 26 as well as their reactivity toward certain electrophiles will be discussed in detail. Alkylation with Alk(3)O(+)BF(4)(-) preferably results in formation of the 3-alkoxy-1lambda(4)-2,6-thiadiazines 29. In the presence of acid or base, also the N-2-alkyl isomers 30 are formed. Mesylation of 26 with methanesulfonic acid anhydride followed by aminolysis furnishes a variety of the desired heterocycles 5 in moderate overall yield. PMID- 12737560 TI - Highly stereocontrolled synthesis of natural barbacenic acid, novel bisnorditerpene from Barbacenia flava. AB - Barbacenic acid, a bisnorditerpene with five contiguous asymmetric centers (four fully substituted), has been prepared for the first time through a highly stereocontrolled route in 5.2% overall yield from a known octalone. The synthesis serves to define the absolute configuration of the natural product. PMID- 12737561 TI - Synthesis of the C(21)-C(26) fragment of superstolide A: concerning the stereochemistry of (E)-crotylboration reactions of alaninal derivatives. AB - A stereoselective synthesis of the C(21)-C(26) fragment of superstolide A has been completed. The absolute and relative stereochemistry of intermediate 14 has been conclusively proven by NMR and X-ray diffraction methods. In the course of this work, it was found that the stereochemistry of 3 had been misassigned in our previously reported synthesis of the C(18)-C(26) segment. This error stems from the unexpected diastereoselectivity in the double asymmetric reaction of N-acetyl d-alaninal 1 and the tartrate ester modified (E)-crotylboronate (R,R)-2. PMID- 12737563 TI - Enhanced stereoselectivity in internucleotidic bond formation by the use of the chiral ribose moiety of thymidine. AB - This paper deals with the synthesis of new cyclic thymidine 3'-phosphoramidite building blocks having a covalent linker between the trityl type 5'-hydroxyl protecting group and the phosphorus atom attached to the 3'-hydroxyl group of thymidine. The ring structures were designed to reduce the conformational freedom around the phosphorus center so that the stereoselectivity in the internucleotide linkage formation would be improved. The linkers were also designed to be removed readily by treatment with aqueous ammonia. These building blocks were synthesized in good yield by one-pot cyclization of the diol precursors with dichloro(N,N diisopropylamino)phosphine, despite their large-membered ring. Various activators having 1H-tetrazole, imidazole, and triazole structures were investigated to find the best selectivity in the synthesis of thymidylyl(5'-3')thymidine phosphorothioate. It turned out that our cyclic phosphoramidites gave preferentially the R(p) diastereoisomer in high coupling yield applicable to the solid-phase synthesis of oligodeoxynucleotides. It should be noted that high stereoselectivity was achieved without any chiral sources other than the 2' deoxyribose moiety itself. The mechanistic studies revealed the importance of the steric bulk and the acidity of the activators. It was also found that the steric bulk of the alcoholic nucleophile was an important factor that determined the stereoselectivity in our systems. PMID- 12737562 TI - The design of novel N-4'-pyridinyl-alpha-methyl proline derivatives as potent catalysts for the kinetic resolution of alcohols. AB - A novel family of chiral acylation catalysts based on a N-4'-pyridinyl-alpha methyl proline structure has been studied. A set of 31 compounds has been easily prepared and screened in the kinetic resolution of racemic alcohol 33 resulting in high enantioselectivities in most cases. From results obtained, H-bonding interactions between the catalyst and the substrate would appear essential to afford high enantioselectivity during the catalytic acylation. Additional solvent dependence and anhydride studies have been made to better identify the mechanism. This work has been further extended to the study of a number of structurally different alcohols. Ethanolamine derivatives in particular were found to be highly effective substrates (up to S = 18.8) in the kinetic resolution. PMID- 12737564 TI - A combination chemical and enzymatic approach for the preparation of azole carboxamide nucleoside triphosphate. AB - Alternative substrates for DNA and RNA polymerases offer an important set of biochemical tools. Many of the standard methods for nucleoside triphosphate synthesis fail in the cases of nonpurine and nonpyrimidine nucleosides. An efficient preparation of the 5'-O-tosylates for both the deoxy- and ribonucleosides enabled preparation of the diphosphate esters by displacement with tris(tetra-n-butylammonium) pyrophosphate. Enzymatic synthesis of the azole carboxamide deoxyribonucleoside triphosphate was based on ATP as the phosphate donor, nucleoside diphosphate kinase as the catalyst, coupled with phosphoenol pyruvate (PEP) and pyruvate kinase as an ATP regeneration system. Ribonucleoside triphosphate synthesis required PEP as the phosphate donor and pyruvate kinase as the catalyst. An optimized purification procedure based upon boronate affinity gel was developed to yield highly purified nucleoside triphosphates. The strategy outlined here provides a new and efficient method for preparation of nucleoside 5'-triphosphate and is likely applicable to a broad variety of base and sugar modified nucleoside analogues. PMID- 12737565 TI - Psammaplins from the sponge Pseudoceratina purpurea: inhibition of both histone deacetylase and DNA methyltransferase. AB - Four novel bisulfide bromotyrosine derivatives, psammaplins E (9), F (10), G (11), and H (12), and two new bromotyrosine derivatives, psammaplins I (13) and J (14), were isolated from the sponge Pseudoceratina purpurea, along with known psammaplins A (4), B (6), C (7), and D (8) and bisaprasin (5). The structures of psammaplins E (9) and F (10), which each contain an oxalyl group rarely found in marine organisms, were determined by spectroscopic analysis. Compounds 4, 5, and 10 are potent histone deacetylase inhibitors and also show mild cytotoxicity. Furthermore, compounds 4, 5, and 11 are potent DNA methyltransferase inhibitors. The biogenetic pathway previously proposed for the psammaplins class is also revisited. PMID- 12737567 TI - Origin of the synchronicity on the transition structures of polar Diels-Alder reactions. Are these reactions [4 + 2] processes? AB - The transition structures (TSs) for a series of related Diels-Alder reactions between cyclopentadiene and mono-, di-, tri-, and tetracyanoethylene derivatives have been studied with use of DFT methods at the B3LYP/6-31G computational level. The increase of the electron-withdrawing substitution on ethylene increases the rate of these polar cycloadditions. However, the symmetric arrangement of cis and trans 1,2-di- and tetracyanoethylenes decreases the effectiveness of the substitution, which can be related to the symmetry found at the corresponding TSs. A DFT analysis of the global and local electrophilicity power of these series of cyano ethylenes provides a sound explanation about the nature of these synchronous processes. The present theoretical study is in agreement with the experimental outcomes. PMID- 12737566 TI - Stereoselective syntheses of 1,4-dideoxy-1,4-imino-octitols and novel tetrahydroxyindolizidines. AB - A new route for the preparation of four new indolizidines, (1R,2S,6S,7S,8aS)- and (1R,2S,6R,7R,8aS)-1,2,6,7-tetrahydroxyindolizidine (30 and 32) and (1S,2R,7S,8S,8aR)- and (1S,2R,7R,8R,8aR)-1,2,7,8-tetrahydroxyindolizidine (44 and 46), is reported. The synthesis is based on Knoevenagel homologation of the readily available enantiomerically pure pyrrolidin-carbaldehydes 13 and 37followed by asymmetric dihydroxylation of the subsequent alkenyl pyrrolidines and cyclization of the corresponding imino-octitols. The new indolizidines and their precursors (imino-octitols 20, 25, 26) and indolizidinones 28a and 28b have been tested for inhibitory activities toward 26 glycosidases. The enzymatic inhibition of trans-7-hydroxy-d-(-)-swainsonine (44) toward alpha-mannosidases is similar to that described for trans-7-hydroxy-l-(+)-swainsonine (11b) toward naringinase (alpha-l-rhamnosidase from Penicillium decumbens). PMID- 12737568 TI - New syntheses and chemistry of hexafluorotropone. AB - Two new routes to hexafluorotropone have been developed, one from hexachlorotropone and a superior synthesis from hexafluorobenzene. Hexafluorotropone was found to be a very weak base, with a conjugate acid pK(a) of -6.2 +/- 0.5. The tropone adds in [6 + 4] fashion to cyclopentadiene and photocyclizes to hexafluorobicyclo[3.2.0]hepta-3,6-dien-2-one. Lithium hydroxide in benzene transforms the tropone into pentafluorotropolone, which functions as a bidentate ligand. PMID- 12737569 TI - Tert-butyl-substituted tripyrranes: insights into the steric and conformational factors that influence porphyrinoid ring formation in the "3 + 1" methodology. AB - The MacDonald "3 + 1" route for porphyrinoid synthesis involves the acid catalyzed condensation of tripyrranes with monocyclic dialdehydes, followed by an oxidation step. In the present study, yields were found to be greatly diminished when tert-butyl substituents were introduced on to the tripyrrane unit. Analysis of the proton NMR spectra for the tripyrranes indicates that the preferred conformation in solution has been radically altered by the presence of these tert butyl moieties. This appears to be the first time that the NMR properties of an intermediate in porphyrin or porphyrin analogue synthesis have been correlated to its effectiveness in macrocycle formation. PMID- 12737570 TI - Study of the lithiated phenylacetonitrile monoanions and dianions formed according to the lithiated base used (LHMDS, LDA, or n-BuLi). 1. Evidence of heterodimer ("Quadac") or dianion formation by vibrational spectroscopy. AB - It is evidenced through vibrational spectroscopy that a heterodimer or "Quadac" is formed when an excess of base (LHMDS, LDA, or n-BuLi) is added to PhCH(2)CN in THF, THF-hexane, or THF-toluene solution. The amount of heterodimer increases with the pK(H)(a) of the lithiated base. A dianionic species may be formed through decomposition of this heterodimer if the pK(H)(a) of the base is sufficiently high, as in the case of n-BuLi. With LDA, only a very small amount of dianion is observed, and with LHMDS, no dianion is detected. The predominant dianionic species observed are the linear and bridged separated ion pairs of the dilithiated dianion. The presence of the amine in the medium is of paramount importance. The PhCHCNLi monomer-dimer equilibrium is entropy driven toward the dimer solvated by the amine. PMID- 12737571 TI - Fluoroalkylation of porphyrins: synthesis and reactions of beta fluoroalkyltetraarylporphyrins. AB - Treatment of 5,10,15,20-tetraarylporphyrins (1) with perfluoroalkyl iodides (2) in the presence of Na(2)S(2)O(4)/NaHCO(3) in DMSO-CH(2)Cl(2) at 30-40 degrees C for several hours gives the corresponding 2-perfluoroalkylporphyrins (3). Nucleophilic attack on 3 with dimethyl malonate, diethyl malonate, malonitrile, or cyano acetate (Nu) anion results in the formation of (E)-3-Nu-2 perfuoroalkyl(methylenyl)chlorins. Electrophilic substitution on 3 with NBS or NO(2) affords regioselectively the corresponding 12(or 13)-bromo- and 12,13 dibromo- or nitroporphyrins. PMID- 12737572 TI - On the regiochemistry of nucleophilic attack on 2-halo pi-allyl complexes. 4. The effect of silver acetate and nucleophile concentrations in competitive nucleophilic attack with malonate and phenoxide nucleophiles. AB - 2,3-Dibromo-1-propene or its allyl carbonate analogue are ionized under Pd catalysis to generate the 2-bromo Pd-pi-allyl complex (triphenylphosphine ligand), which alkylates with malonate nucleophile at the terminal position. The presence of acetate ion in the reaction mixture results in some malonate attack being redirected to the central carbon. The acetate ion can come from the ionization of 1-acetoxy-2-bromo-2-propene or by the addition of silver acetate to the reaction mixture. The addition of phenoxide ion to the reaction also causes the same regiochemical phenomena, although harder anions such as methoxide exert no such effect. PMID- 12737573 TI - Asymmetric hydrogenations for the synthesis of Boc-protected 4-alkylprolinols and prolines. AB - The utility of 4-substituted prolinols and their corresponding prolines in peptides, peptidomimetics, and natural products has motivated researchers to find new and efficient routes for their preparation. Herein, we report a general approach to the synthesis of Boc-protected 4-alkylprolinols and prolines via a divergent asymmetric hydrogenation strategy. Intermediate exocyclic olefins were prepared by Wittig-type reactions with ketone 6 and subjected to hydroxyl and sterically directed reductions. The Crabtree catalyst (Ir[COD]PyPCy(3)PF(6)) proved to be highly effective in diastereoselective hydrogenations to give trans substituted pyrrolidines (9). Good facial selectivities were also observed in heterogeneous hydrogenations with Raney-nickel to obtain cis-substituted pyrrolidines (11). Employing this strategy, we describe the synthesis of novel prolinol and proline-based building blocks for incorporation into biologically relevant peptidomimetics. PMID- 12737574 TI - Ionic reaction of halogens with terminal alkenes: the effect of electron withdrawing fluorine substituents on the bonding of halonium ions. AB - Ionic reactions of terminal alkenes with chlorine (Cl(2)), bromine (Br(2)), and iodine monochloride (ICl) are sensitive to the alkyl substituents, and the positions and number of vinyl fluorine atoms. These perturbations influence the symmetry of the halonium ion intermediates, which can be determined by the distribution of the Markovnikov to anti-Markovnikov products. A vinyl fluorine on the number-2 carbon favors an unsymmetrical intermediate with greater charge on the number-2 carbon unless the alkyl group is electron withdrawing. A vinyl fluorine on the terminal number-1 carbon favors positive charge development on that carbon unless a resonance stabilizing group is on the number-2 carbon. The symmetry of halonium ions with vinyl fluorines on both carbons-1 and -2 depends primarily on the characteristics of the alkyl substituent. Intermediates range from open-ions with the positive charge on carbon-2, to various bridged species, to open-ions on the terminal carbon. PMID- 12737575 TI - Palladium-catalyzed tetrakis(dimethylamino)ethylene-promoted reductive coupling of aryl halides. AB - Tetrakis(dimethylamino)ethylene (TDAE)/cat. PdCl(2)(PhCN)(2)-promoted reductive coupling of aryl bromides having either electron-donating or electron-withdrawing groups on their para- and/or meta-position proceeded smoothly to afford the corresponding biaryls in good to excellent yields. Notably, TDAE is such a mild reductant that easily reducible groups, such as carbonyl and nitro groups, are tolerate. A similar reductive coupling of ortho-substituted aryl bromides did not occur at all. The proper choice of palladium catalysts is essential for the reductive coupling; thus, PdCl(2)(PhCN)(2), PdCl(2)(MeCN)(2), Pd(hfacac)(2), Pd(2)(dba)(3), PdCl(2), and Pd(OAc)(2) were used successively for this reaction, but phosphine-ligated palladium catalysts such as Pd(PPh(3))(4), PdCl(2)(PPh(3))(2), and Pd(dppp) did not promote the reaction. The reductive coupling did not occur with nickel catalysts such as NiBr(2), NiCl(2)(bpy), and Ni(acac)(2). The TDAE/cat. palladium-promoted reductive coupling of aryl halides having electron-withdrawing groups took place more efficiently than that of aryl halides substituted with electron-donating groups. A plausible mechanism of TDAE/cat. palladium-promoted reaction is discussed. PMID- 12737576 TI - Omega-halonitriles: domino cyclizations to oxa- and carbocyclic nitriles. AB - t-BuOK-induced deprotonation of omega-haloalkylnitriles generates remarkably stable potassiated nitriles. In situ deprotonation and alkylation of omega chloroalkylnitriles with aldehyde electrophiles trigger sequential nucleophilic electrophilic alkylations generating substituted tetrahydrofuranyl and tetrahydropyranyl nitriles. Redirecting the cyclization manifold with 5 iodopentanenitrile and a ketone causes a complementary electrophilic-nucleophilic cyclization to the corresponding carbonitrile. Collectively these cyclizations provide rapid assembly of five- and six-membered oxa- and carbocyclic nitriles demonstrating the utility of omega-halonitriles in domino alkylations. PMID- 12737577 TI - Molecular recognition of NO/NO+ via multicenter (charge-transfer) binding to bridged diarene donors. Effect of structure on the optical transitions and complexation thermodynamics. AB - Bridged diarenes form very strong [1:1] complexes with nitrosonium/nitric oxide in which the NO moiety is optimally sandwiched in the cleft between a pair of cofacial aromatic rings which act as a molecular "Venus flytrap". The spectral features of these associates are generally similar to those for [1:1] and [2:1] nitrosonium complexes with mononuclear alkyl-substituted benzenes, and they are appropriately described within the LCAO molecular-orbital methodology and the Mulliken (charge-transfer) formulation of donor/acceptor electronic transitions. The thermodynamics study indicates that the efficient binding is determined by (i) the close matching of the donor/acceptor redox potentials and (ii) the ability of bridged diarenes for multicentered interactions with a single NO moiety. The best fit of the electronic and structural parameters is provided by a calixarene host that allows the interacting centers to be arranged in a manner similar to those extant in [2:1] nitrosonium complexes with analogous (nonbridged) aromatic donors; this results in its very strong noncovalent binding with nitrosonium/nitric oxide with the formation constant of K(B) approximately 10(8) M(-)(1) and free-energy change of -DeltaG degrees = 45 kJ mol(-)(1). Such strong, selective, and reversible bindings of nitrosonium/nitric oxide by (cofacial) aromatic centers thus provide the basis for the development of efficient NO sensors/absorbents and also suggest their potential relevance to biochemical systems. PMID- 12737578 TI - A theoretical study of reactivity and regioselectivity in the hydroxylation of adamantane by ferrate(VI). AB - The conversion of adamantane to adamantanols mediated by ferrate (FeO(4)(2)(-)), monoprotonated ferrate (HFeO(4)(-)), and diprotonated ferrate (H(2)FeO(4)) is discussed with the hybrid B3LYP density functional theory (DFT) method. Diprotonated ferrate is the best mediator for the activation of the C-H bonds of adamantane via two reaction pathways, in which 1-adamantanol is formed by the abstraction of a tertiary hydrogen atom (3 degrees ) and 2-adamantanol by the abstraction of a secondary hydrogen atom (2 degrees ). Each reaction pathway is initiated by a C-H bond cleavage via an H-atom abstraction that leads to a radical intermediate, followed by a C-O bond formation via an oxygen rebound step to lead to an adamantanol complex. The activation energies for the C-H cleavage step are 6.9 kcal/mol in the 1-adamantanol pathway and 8.4 kcal/mol in the 2 adamantanol pathway, respectively, at the B3LYP/6-311++G level of theory, whereas those of the second reaction step corresponding to the rebound step are relatively small. Thus, the rate-determining step in the two pathways is the C-H bond dissociation step, which is relevant to the regioselectivity for adamantane hydroxylation. The relative rate constant (3 degrees )/(2 degrees ) for the competing H-atom abstraction reactions is calculated to be 9.30 at 75 degrees C, which is fully consistent with an experimental value of 10.1. PMID- 12737579 TI - Reaction of alpha-oxoketene-N,S-arylaminoacetals with Vilsmeier reagents: an efficient route to highly functionalized quinolines and their benzo/hetero-fused analogues. AB - A simple, highly efficient, and regioselective synthesis of functionalized quinolines through Vilsmeier cyclization of a variety of alpha-oxoketene-N,S anilinoacetals has been reported. The cyclization is found to be facile with N,S acetals bearing strongly activating groups on aniline, whereas yields of quinolines are moderate in other cases. The reaction could also be extended for the synthesis of substituted tricyclic benzo[h]quinoline, pyrido[2,3-h]quinoline, 4,7-diphenylphenanthroline, and tetracyclic quino[8,7-h]quinoline by performing a Vilsmeier reaction on N,S-acetals derived from 1-naphthylamine, m phenylenediamine, o-phenylenediamine, and 1,5-diaminonaphthalene, respectively. A few of the newly synthesized quinolines are subjected to further transformation to afford 2-unsubstituted (Raney-Ni/Ethanol), quinoline-5,8-quinone (NBS/H(2)SO(4)), or 2-alkyl/aryl aminoquinolines through sequential m-CPBA oxidation to the corresponding (2-methylsulfonyl)quinoline followed by replacement with appropriate amines. Similarly, cycloannulation of a few 2 methylthio-3-benzoylquinolines with hydrazine hydrate under microwave irradiation afforded the corresponding substituted and fused pyrazolo[3,4-b]quinolines in excellent yields, whereas TBTH/AIBN-mediated cyclization of the corresponding 3 (2-bromobenzoyl)-2-methylthioquinolines yielded the corresponding benzothiopyrano fused quinolines through radical translocation. PMID- 12737580 TI - Enantioconvergent synthesis of (-)-(2R,5S)-1-allyl-2,5-dimethylpiperazine, an intermediate to delta-opioid receptor ligands. AB - A convenient, high-yield enantioconvergent synthesis of (-)-1-allyl-(2S,5R) dimethylpiperazine from trans-2,5-dimethylpiperazine has been developed. This compound is an important intermediate in the synthesis of delta-opioid receptor ligands. The process allows for the laboratory preparation of 100 g quantities of this enantiomerically pure diamine without chromatography. The key steps in the sequence were an efficient optical resolution using relatively inexpensive resolving agents, followed by interconversion of the unwanted (+)-enantiomer into the desired (-)-enantiomer. PMID- 12737581 TI - Intramolecular [2 + 2] photocycloaddition of alkenes incorporated in a carbohydrate template. Synthesis of enantiopure bicyclo[3.2.0]heptanes and [6.3.0]undecanes. AB - Intramolecular [2 + 2] photocycloaddition of alkenes with a furano sugar placed between them have been investigated under both copper(I)-catalyzed and sensitized conditions. The copper(I)-catalyzed photocycloaddition of the dienes 4a, 4b, and 4c led to unexpected formation of the thermodynamically less stable cis-syn-cis 4 5-5 tricyclic adducts 5a, 5b, and 5c, respectively. The sensitized photocycloaddition of the diene 14 also gave the cis-syn-cis adduct 15 showing that the copper(I) catalyst does not have any influence on the stereochemical course through coordination with the anomeric ring oxygen of the furano sugar. The identical stereochemical course observed under both catalyzed and sensitized photoaddition reactions have been attributed to be of steric origin. Bis(dienes) 25a and 25b, which gave an intractable mixture on copper(I)-catalyzed irradiation, underwent smooth photocycloaddition in the presence of benzophenone, and the resulting 1,2-divinyl cyclobutanes underwent spontaneous [3.3] rearrangement at room temperature to produce bicyclo[6.3.0]undecanes 30a and 30b, respectively. This investigation provides an approach for the construction of enantiopure bicyclo[3.2.0]heptanes and -[6.3.0]undecanes. PMID- 12737582 TI - Dications of fluorenylidenes. The effect of substituent electronegativity and position on the antiaromaticity of substituted tetrabenzo[5.5]fulvalene dications. AB - Oxidation of 3,6-disubstituted tetrabenzo[5.5]fulvalenes by SbF(5) results in the formation of dications that behave like two antiaromatic fluorenyl cations connected by a single bond. Both fluorenyl systems exhibit the paratropic shifts and nucleus independent chemical shifts (NICS) characteristic of antiaromatic species. Comparison with analogous 2,7-disubstituted tetrabenzo[5.5]fulvalenes reveals that the antiaromaticity of the substituted ring system can be altered substantially by changes in the placement of the substituents, possibly due to changes in the delocalization of charge in the system. Substituents in the 3,6 position decrease the antiaromaticity because of the increase in the benzylic resonance compared to 2,7-substituents. PMID- 12737583 TI - Synthesis of metal-chelating lipids to sensitize lanthanide ions. AB - Sensitization of lanthanide ions is important for lanthanide ion-based assays and sensing. To the best of our knowledge, there are very few reports of lanthanide ion sensitization after it is incorporated into the liposome surface. This paper describes the syntheses of several saturated and polymerizable metal-chelating lipids based on chelidamic acid. The lipids are synthesized either from (S) ornithine or racemic 2,3-diaminopropanoic acid. These lipids as well as polymerized liposomes incorporating these lipids sensitize lanthanide ions. Liposomes from the lipid 18-Eu(3+) provided a probe that relies not only on the emission wavelengths of Eu(3+) but also on a reproducible lifetime that can be used for protein identification. PMID- 12737584 TI - Synthesis of (+)-manoalide via a copper(I)-mediated 1,2-metalate rearrangement. AB - An enantiospecific synthesis of the phospholipase A(2) inhibitor (+)-(4R) manoalide is reported in which all 25 carbons of the sesterterpenoid skeleton are constructed from 3-furaldehyde, trimethylalane, oxirane, CO, beta-ionone, and propargyl bromide. The overall yield for the longest linear sequence (12 steps) is 12%. Key steps include (a) a zirconium-catalyzed carboalumination reaction to construct the C10-C11 trisubstituted alkene, (b) a Cu(I)-mediated 1,2-metalate rearrangement to construct the C6-C7 trisubstituted alkene, (c) a Sharpless kinetic resolution to secure the (4R)-stereochemistry, (d) generation of a 5 stannyl-2,3-dihydrofuran by Mo-catalyzed cycloisomerization of a homopropargylic alcohol, and (e) construction of the hydroxyfuranone ring by photooxidation of a silylfuran. PMID- 12737585 TI - Synthesis and characterization of a metallocycle-based molecular shuttle. AB - Kinetically stable metallocycle-based molecular shuttles of [2]rotaxanes 4a and 4b, along with [3]rotaxanes 5a and 5b, have been prepared using the rhenium(I) bridged metallocycle 2 and the dumbbell components containing two stations, 3a and 3b. The rotaxanes were self-assembled by hydrogen bonding interactions upon heating a Cl(2)CHCHCl(2) solution containing their components at 70 degrees C. Each rotaxane was isolated in pure form by silica gel chromatography under ordinary laboratory conditions and fully characterized by elemental analysis and various spectroscopic methods. The (1)H NMR signals for the amide NH and the methylene -(CH(2))(4)- of the station were considerably changed when occupied by the metallocycle. In [2]rotaxane 4b, which has a larger naphthyl spacer, the occupied and unoccupied stations gave widely separated signals in the (1)H NMR spectroscopy at room temperature, but averaged signals of two stations were observed in [2]rotaxane 4a, which has a smaller phenyl spacer. This is attributed to the shuttling of the metallocycle between two stations. The coalescence temperature experiment gave a shuttling rate of approximately 670 s(-)(1) at 19 degrees C in CDCl(3), corresponding to an activation free energy (DeltaG()) of 13.3 kcal/mol. With respect to the relative position of the chloride in the rhenium(I) center, two diastereomers are possible in the [2]rotaxane and three diastereomers are possible in the [3]rotaxane. In fact, the rotaxanes exist as diastereomeric mixtures in nearly equal amounts of all possible diastereomers on the basis of the amide NH signals of the station in the (1)H NMR spectroscopy. PMID- 12737586 TI - First total synthesis of a GPI-anchored peptide. AB - A GPI-anchored dipeptide of sperm CD52 antigen was prepared through a convergent synthesis. First, the dipeptide with its C-terminus free and the GPI with its nonreducing end phosphoethanolamine bearing a free amino group were synthesized separately. Then, the two building blocks were coupled with use of EDC/HOBt as the condensation reagent. Finally, the GPI-anchored peptide was deprotected to give the target molecule 1. PMID- 12737587 TI - Triethylaluminum- or triethylborane-induced free radical reaction of alkyl iodides and alpha,beta-unsaturated compounds. AB - A series of alpha,beta-unsaturated compounds, 1a-c, 9, 13, and 17, were used as reactants in free radical conjugate addition reactions with different radicals generated from alkyl iodides such as 3, 4, or 5 in the presence of triethylborane oxygen in air or via the use of triethylaluminum-benzoyl peroxide as a free radical initiator. When the reactions were carried out using triethylborane-air, the products, in most cases, were clean and were easily purified. However, higher yields of the 1,4-adducts and less side reactions occurred when less reactive substrates were used as Michael acceptors in reactions with triethylaluminum benzoyl peroxide and alkyl iodide under similar conditions. A mechanism for this is proposed in Scheme 1. PMID- 12737588 TI - Total synthesis of (+)-boronolide, (+)-deacetylboronolide, and (+) dideacetylboronolide. AB - Total synthesis of (+)-boronolide, (+)-deacetylboronolide, and (+) dideacetylboronolide has been achieved from a single intermediate 26, which was synthesized in 11 steps from a d-mannitol-derived intermediate 8 in an overall yield of 10%. The key steps in the synthesis are inversion of a chiral center by taking an advantage of the inherent mechanism involved in the ring closing to an epoxide via intramolecular S(N)2 reaction and lactonization of a diol using Fetizons reagent. The strategy is amenable to preparation of analogues of (+) boronolide in sufficient amount for further screening of biological activity. PMID- 12737589 TI - Nucleophilic displacements in supercritical carbon dioxide under phase-transfer catalysis conditions. 2. Effect of pressure and kinetics. AB - The nucleophilic displacement on n-octylmesylate (n-C(8)H(17)OSO(2)CH(3), 1) with four different anions (I(-), Br(-), N(3)(-), and SCN(-)) is investigated under liquid-supercritical phase-transfer catalysis (LSc-PTC) conditions, i.e. in a biphase system of supercritical carbon dioxide (scCO(2)) and water, in the presence of both silica supported and conventional onium salts. The CO(2) pressure greatly affects the concentration of 1 in the sc-phase and plays a major role on its conversion. For example, at 50 degrees C and with a supported PT catalyst, the conversion of 1 into n-octyl iodide drops by a factor of 5 as the CO(2) pressure is increased from 80 to 150 bar, while in the same pressure range, the solubility of n-octylmesylate in scCO(2) shows a 6-fold increase, indicating that the reagent is desorbed from the catalyst. Under LSc-PTC conditions, pseudo first-order kinetic rate constants, evaluated for the investigated reactions, show that the performance of scCO(2) as a PTC solvent and the relative nucleophilicity order of the anions (N(3)(-) > I(-) > or = Br(-) > SCN(-)) are comparable to those of toluene and n-heptane. The behavior of conventional phosphonium salts in the scCO(2)/H(2)O biphase system suggests that the reaction may take place either within small droplets of PT-catalyst containing water or in a separate third liquid phase of the PT-catalyst itself. PMID- 12737590 TI - Activity difference between alpha-COOH and beta-COOH in N-phosphorylaspartic acids. AB - N-phosphorylamino acids are chemically active species that have many biomimic activities. alpha-COOH in amino acids and peptides behaviors rather differently than beta-COOH in many biochemical processes and takes a more important role in the origin of life. Activity differences between alpha-COOH and beta-COOH in the peptide formation of phosphoryl amino acids are studied by 1D, 2D NMR techniques and by ab initio and density functional theory (DFT) calculations in this paper. Phosphoryl dipeptide is formed directly from phosphoryl aspartic acids without any coupling reagents. Only the alpha-dipeptide ester is observed by 1D (1)H, (13)C, and (31)P NMR and 2D NMR. In the ab initio and DFT calculations, the pentacoordinate phosphorane intermediates containing five-membered rings are predicted to be more favored than those with six-membered rings. Both the experimental results and the theoretical calculations suggest that only the alpha COOH group is activated by N-phosphorylation in N-phosphorylaspartic acid under mild conditions. PMID- 12737591 TI - A DFT study on the regioselectivity of the reaction of dichloropropynylborane with isoprene. AB - This theoretical study deals with the reaction of isoprene and dichloropropynylborane. We report the results of the DFT calculations applied to the two processes involved, Diels-Alder cycloaddition and 1,4-alkynylboration. The boron influences both the chemoselectivity and the regioselectivity of this reaction through secondary orbital interactions (SOI hereafter) that give rise to transition structures with strong [4 atom + 3 atom] character. The "meta" regioselectivity observed experimentally for the reaction between 2-substituted 1,3-dienes and alkynyldihaloboranes has been explained as a result of the higher stabilization of these transition structures with "meta" orientation. Intrinsic reaction coordinate calculations were performed to determine connectivities and established the remarkable result that the geometrically very similar transition structures leading to both regioisomeric 1,4-alkynylboration products correspond to different pathways. For the "meta" orientation a direct alkynylboration of the diene through a concerted transition structure was found. PMID- 12737592 TI - The first highly enantioselective homogeneously catalyzed asymmetric reductive amination: synthesis of alpha-N-benzylamino acids. AB - High-throughput screening considering a library of 96 chiral P-ligands involved in two types of Rh(I) complexes was used for the identification of homogeneous catalysts for the highly enantioselective reductive amination of alpha-keto acids with benzylamine. After optimization of the reaction conditions and scale-up with a cationic Rh-Deguphos catalyst, a range of chiral alpha-amino acids could be produced by this new reaction in good yield and by up to 98% ee. PMID- 12737593 TI - A practical one-pot synthesis of soluble hexa-peri-hexabenzocoronene and isolation of its cation-radical salt. AB - A simple and practical synthesis of soluble hexa-peri-hexabenzocoronene (HBC) from readily available hexaphenylbenzene (HPB) is described. In this simple procedure, the substitution of the free para positions of the propeller-shaped HPB with tert-butyl groups and the oxidative cyclodehydrogenation to planar HBC is achieved in a one-pot reaction using ferric chloride both as a Lewis acid catalyst and as an oxidant in excellent yields. The ready availability of HBC allows the isolation of its pure cation-radical salt using a variety of chemical oxidants such as antimony pentachloride and triethyloxonium and nitrosonium hexachloroantimonate salts. PMID- 12737595 TI - Cobalt-catalyzed one-pot three-component coupling route to beta-acetamido carbonyl compounds: a general synthetic protocol for gamma-lactams. AB - An efficient improved procedure for the synthesis of beta-acetamido carbonyl compounds is developed by a cobalt(II) chloride-catalyzed three-component coupling protocol. The procedure is also amenable to the synthesis of gamma lactams by a three-component coupling reaction with use of 2 carbomethoxybenzaldehyde. The beta-acetamido carbonyl compounds derived from 2 carbomethoxybenzaldehyde are useful intermediates as they can be transformed to the corresponding gamma-lactams on treatment with base. PMID- 12737596 TI - Copper(II)-catalyzed [2,3]-sigmatropic rearrangement of N methyltetrahydropyridinium ylids. AB - A ring-contractive and highly diastereoselective [2,3]-sigmatropic rearrangement occurs when N-methyl-1,2,3,6-tetrahydropyridine is treated with sub stoichiometric amounts of copper or rhodium salts, in the presence of ethyl diazoacetate, giving ethyl cis-N-methyl-3-ethenyl proline (4). PMID- 12737594 TI - Meso-tetraaryl cofacial bisporphyrins delivered by Suzuki cross-coupling. AB - The Suzuki cross-coupling methodology provides a facile synthetic approach for the modular preparation of meso-tetraaryl cofacial bisporphyrins anchored by xanthene and dibenzofuran. This synthetic method furnishes cofacial bisporphyrin templates with enhanced steric and electronic protection from mu-oxo formation and oxidative degradation. The ability of these platforms to support multielectron oxidation chemistry mediated by proton-coupled electron transfer (PCET) is demonstrated by their reactivity for the catalytic disproportionation of hydrogen peroxide to oxygen and water. PMID- 12737597 TI - Highly enantioselective oxidations of ketene dithioacetals leading to trans bis sulfoxides. AB - Ketene dithioacetals undergo a Sharpless-type asymmetric oxidation using (+)-DET, Ti(O(i)()Pr)(4), and cumene hydroperoxide to give the trans bis-sulfoxides 4a-f with essentially complete control of enantioselectivity and diastereoselectivity. The high enantioselectivity is a consequence of carrying out two asymmetric processes on the same substrate. However, this should lead to the formation of a small amount of the meso isomer but none was isolated. From monitoring the enantioselectivity of the monoxide over time, it was concluded that small amounts of the meso isomer must be formed. The inability to isolate this compound could be because it acted as a ligand on titanium and remained tightly bound even upon workup. PMID- 12737598 TI - Synthesis of pentacyclic 13-azadibenzo[a,de]anthracenes via anionic cascade ring closure. AB - Bromine-lithium exchange using tert-butyllithium at -78 degrees C initiates a cascade process whereby either xanthone derivatives or pentacyclic 13 azadibenzo[a,de]anthracenes are produced in high yields. The reaction proceeds via a sequential intramolecular trapping of organolithium intermediates. PMID- 12737599 TI - Efficient and regioselective synthesis of 2-alkyl-2H-indazoles. AB - An efficient and regioselective synthesis of 2-methyl-2H-indazoles and 2-ethyl-2H indazoles using trimethyloxonium tetrafluoroborate or triethyloxonium hexafluorophosphate is reported. PMID- 12737600 TI - Highly selective methodology for the direct conversion of aromatic aldehydes to glycol monoesters. AB - Al(2)O(3)/MeSO(3)H (AMA) was found to be an extremely efficient reagent for the conversion of aromatic aldehydes and diols to glycol monoesters. The remarkable selectivity achieved with this reagent is an attractive feature of the present method. PMID- 12737601 TI - Iodomethyl group as a hydroxymethyl synthetic equivalent: application to the syntheses of D-manno-hept-2-ulose and l-fructose derivatives. AB - The one-carbon elongation of aldoses to ketoses using iodomethyllithium as the key reagent in the homologation step is exemplified by the preparation of two carbohydrates of chemical and biological interests: d-manno-hept-2-ulose from d mannose and l-fructose from l-arabinose. PMID- 12737603 TI - Revised structure of a purported 1,2-dioxin: a combined experimental and theoretical study. AB - 3,6-Bis(p-tolyl)-1,2-dioxin (1g) was suggested by Shine and Zhao as a product in an electron-transfer (ET) photochemical reaction. This photoproduct is instead shown to be (E)-1,4-di-p-tolylbut-2-ene-1,4-dione ((E)-4a). Ab initio and DFT calculations indicate that ring-closed 1,2-dioxin is thermodynamically far less stable than open-chain but-2-ene-1,3-dione. These calculations indicate that (E) 4a is formed via the cation radical of 1g, which sequentially isomerizes to a novel sigma-radical with an O,O 3e bond [(Z)-4a](+)(*), undergoes ET to give (Z) 4a, and then photoisomerizes to (E)-4a. PMID- 12737602 TI - Efficient synthesis of [6-chloro-2-(4-chlorobenzoyl)-1H-indol-3-yl]-acetic acid, a novel COX-2 inhibitor. AB - The synthesis of 6-chloro-2-(4-chlorobenzoyl)-1H-indol-3-ylacetic acid (1), a selective cyclooxygenase 2 (COX-2) inhibitor, is described. The synthesis relied on a novel indole formation that involved an alkylation/1,4 addition/elimination/isomerization cascade. It was demonstrated that the entire sequence from sulfonamide 13 and bromoketone 14 to the desired indole (1) could be executed in a single pot. PMID- 12737604 TI - Preparation of bis(beta-trimethylsilylethanesulfonyl)imide and its use in the synthesis of protected amine derivatives. AB - Bis(beta-trimethylsilylethanesulfonyl)imide (SES(2)NH) can be easily prepared in 85% yield by alkylation of the trianion of bismethanesulfonimide with 2 equiv of commerically available (iodomethyl)trimethylsilane. This synthon undergoes effective Mitsunobu alkylation reactions with both primary and secondary alcohols to afford the corresponding bis-SES imides. These imides can be selectively cleaved to the mono-SES-protected amines, and in addition undergo a one-pot cleavage/N-alkylation to afford SES derivatives of secondary amines. PMID- 12737606 TI - Highly enantioselective reductive amination of simple aryl ketones catalyzed by Ir-f-Binaphane in the presence of titanium(IV) isopropoxide and iodine. AB - Using an Ir-f-Binaphane complex as the catalyst, complete conversions and high enantioselectivies (up to 96% ee) were achieved in the asymmetric reductive amination of aryl ketones in the presence of Ti(O(i)()Pr)(4) and I(2). A simple and efficient method of synthesizing chiral primary amines has been realized. PMID- 12737605 TI - Total synthesis of buergerinin F and buergerinin G. AB - Syntheses of buergerinin F (1) and buergerinin G (2) were carried out to establish the absolute stereochemistry of these natural products. A linear sequence was used to synthesize 1 in 15 steps and 9% overall yield from thymidine. Subsequent oxidation of 1 with ruthenium tetroxide afforded 2 in 77% yield. PMID- 12737607 TI - Peptidic alpha-ketocarboxylic acids and sulfonamides as inhibitors of protein tyrosine phosphatases. AB - One common approach for designing protein tyrosine phosphatase (PTPase) inhibitors is to incorporate a nonhydrolyzable phosphotyrosine (pTyr) mimic into a peptide substrate for PTPases. This report describes the synthesis of three such nonhydrolyzable pTyr mimics that contain alpha-ketoacid, alpha-hydroxyacid, and methylenesulfonamide functional groups in place of the phosphate. These pTyr mimics were incorporated into the peptide sequence Ac-Asp-Ala-Asp-Glu-X-Leu NH(2), where X is the pTyr mimic, and analyzed for activity against the Yersinia PTPase and PTP1B. PMID- 12737609 TI - Self-advocacy, "learning difficulties," and the social model of disability. AB - In this paper we draw upon a research project whose authors examined the self advocacy of people with the label of "learning difficulties." We examined self advocacy in action within groups and explored understandings of self-advocacy accounted for in personal narratives. We identified the background of our study, reflected on the use of ethnographic and narrative methods, and pinpointed three findings that highlight the significance of self-advocacy in the lives of people with the label of "learning difficulties": (a) the distributed and multilocated nature of self-advocacy, (b) self-advocacy's dialogue with the relational nature of "learning difficulties," and (c) support for the self-advocacy movement. We conclude that a variety of theoretical, policy, and practical implications surface through an engagement with the self-advocacy movement. PMID- 12737610 TI - Eyewitness identification accuracy: a comparison of adults with and those without intellectual disabilities. AB - The effect of variation in the clarity of a witnessed event on the accuracy of eyewitness identification for adults with intellectual disabilities and those without disabilities was examined. Following observation of one of three films (clear, less distinct, or ambiguous) depicting a nonviolent theft, participants were asked to identify the thief from a photo lineup. Across all film conditions, participants with intellectual disabilities made as many correct identifications as did participants without disabilities, but they also made more false identifications and were more prone to guessing. Differences between groups seemed to be attributable to the demand factors inherent in the eyewitness identification task and understanding of the nature of the task itself. PMID- 12737612 TI - Supported employment outcomes across a decade: is there evidence of improvement in the quality of implementation? AB - There is little question that the strategies used to improve supported employment outcomes, namely higher wages and higher levels of integration, have changed since the mid-1980s. Innovations of natural supports and employer leadership have helped increase the capacity of provider agencies and the business community to include people with disabilities in the workforce. This report is the sixth in a series that focuses on features of natural supports and its relationship to outcomes. Our purpose in this paper is to describe an analysis designed to investigate the features of employment, wage, and integration outcomes of jobs acquired by people with disabilities early in the development of supported employment compared to more recent years. PMID- 12737611 TI - Family support in the United States: financing trends and emerging initiatives. AB - Trends in family support spending and new programmatic initiatives across the country during the 1990s are summarized. Nationally, total spending for family support exceeded $1 billion in 2000. Between 1990 and 2000, spending for such services in the United States grew from 1.5% to 4% of total resources expended by state MR/DD agencies. However, we found considerable state variability in level of resources allocated to support families providing care to a member with developmental disabilities. Programmatic initiatives include trends toward family directed services, targeted programs for special populations, such as ethnic minorities and aging caregivers, and the slow expansion of cash subsidies. Recommendations for the future course of policy in this area are provided. PMID- 12737613 TI - Establishing mental retardation in capital cases: an update. PMID- 12737614 TI - Toward a uniform standard for mental retardation in death penalty cases. PMID- 12737615 TI - Treatment or accommodation for adults with challenging behaviors. PMID- 12737616 TI - Seeing with new eyes: metaphors of family experience. PMID- 12737619 TI - Effect of repetitive exercise testing on breathlessness in humans. AB - There are conflicting reports on the reproducibility of the visual analogue scale (VAS) and the modified Borg scale for the estimation of breathlessness during exercise. In an attempt to clarify the situation, two groups of healthy subjects undertook a progressive exercise test either daily (Group A) or weekly (Group B) on 10 separate occasions. Breathlessness was estimated every 1 min using the VAS. After 10 occasions, both Group A (P <0.05) and Group B ( P <0.01) showed a significant increase in the mean intercept of the breathlessness/ventilation (VAS/ V (I)) relationship. The increase was not progressive; using change point regression, reproducible values were found to occur after approximately the fifth occasion in both subject groups. As the slope of the VAS/ V (I) relationship was highly reproducible and did not change with repeat testing, it would appear that at least two mechanisms are involved in the generation of the sensation of breathlessness. A decrease in the exercise heart rate over the same time period was significantly correlated with changes in the VAS/ V (I) intercept in both groups (P <0.01 and P <0.005 respectively). The relationship is unlikely to be causal, but may be indicative of a common underlying mechanism. It is suggested that breathlessness scores are likely to decrease as a direct result of repetitive testing over, on average, the first five periods of assessment. On the basis of this study, it may be inferred that a physiological mechanism contributes to the modulation of breathlessness during repetitive exercise testing. PMID- 12737620 TI - MHC class I receptors on natural killer cells: on with the old and in with the new. AB - The functions of natural killer (NK) cells are controlled by an abundance of activating and inhibitory receptors. Many of these interact with MHC class I molecules. The MHC system also interacts with cytotoxic T-lymphocytes and has been shown to comprise a rapidly evolving family of molecules. This challenges the functional relationship of NK cell receptors with their ligands. Although individual receptors have become subject to species-specific expansions over evolutionary time, the main themes of the NK cell interaction with MHC class I have been preserved. This review details the interaction of NK cell receptors with MHC class I and discusses their unexpectedly rapid evolution. PMID- 12737622 TI - Correction of an enzyme trafficking defect in hereditary kidney stone disease in vitro. AB - In normal human hepatocytes, the intermediary-metabolic enzyme alanine:glyoxylate aminotransferase (AGT) is located within the peroxisomes. However, in approx. one third of patients suffering from the hereditary kidney stone disease primary hyperoxaluria type 1, AGT is mistargeted to the mitochondria. AGT mistargeting results from the synergistic interaction between a common P11L (Pro11-->Leu) polymorphism and a disease-specific G170R mutation. The polymorphism generates a functionally weak mitochondrial targeting sequence, the efficiency of which is increased by the mutation. The two substitutions together, but not in isolation, inhibit AGT dimerization, highlighting the different structural requirements of the peroxisomal and mitochondrial protein-import machineries. In the present study, we show that treatments known to increase the stability of proteins non specifically (i.e. lowering the temperature from 37 to 30 degrees C or by the addition of glycerol) completely normalize the intracellular targeting of mutant AGT expressed in transfected COS cells. On the other hand, treatments known to decrease protein stability (e.g. increasing the temperature from 37 to 42 degrees C) exacerbate the targeting defect. Neither of the treatments affects the relative efficiencies of the peroxisomal and mitochondrial protein-import pathways intrinsically. Results are discussed in the light of the known structural requirements of the two protein trafficking pathways and the formulation of possible treatment strategies for primary hyperoxaluria type 1. PMID- 12737621 TI - Increased expression of angiopoietin-2 and Tie2 receptor in a rat model of myocardial ischaemia/reperfusion. AB - The angiopoietins and Tie receptors are involved in blood vessel formation. The role of the angiopoietin/Tie receptor system in myocardial ischaemia/reperfusion is not well known. To investigate the participation of angiopoietins and Tie receptors in myocardial ischaemia/reperfusion, adult Wistar rats were studied in which the left coronary artery was ligated for 30 min, followed by reperfusion. Angiopoietin-1 (Ang1), angiopoietin-2 (Ang2), Tie1 and Tie2 were measured immediately after relief of occlusion, and 1, 6, 24 and 72 h after reperfusion, by Northern blot, Western blot and immunohistochemical staining. Ang2 mRNA was increased significantly at 24 h and 48 h after reperfusion, and returned to baseline levels at 72 h, in the jeopardized myocardium. Tie2 mRNA increased 3.4 fold immediately after the relief of occlusion, reached a maximum 8-fold increase at 24 h after reperfusion and remained elevated up to 72 h. Ang2 protein levels also increased after reperfusion, reaching a maximum 2.2-fold increase at 48 h after reperfusion. Tie2 protein increased immediately after relief of ischaemia, and showed a significant increase from 6 h to 72 h after reperfusion as compared with the sham control. Ang1 and Tie1 mRNA and protein did not show significant changes after ischaemia/reperfusion. Immunohistochemical studies also showed increased immunoreactivity of Ang2 and Tie2 in the jeopardized myocardium after ischaemia/reperfusion. In conclusion, expression of both Ang2 and Tie2 increased after ischaemia/reperfusion in the rat ventricular myocardium, while the expression of Ang1 and Tie1 did not. PMID- 12737623 TI - Subsite specificity (S3, S2, S1', S2' and S3') of oligopeptidase B from Trypanosoma cruzi and Trypanosoma brucei using fluorescent quenched peptides: comparative study and identification of specific carboxypeptidase activity. AB - We characterized the extended substrate binding site of recombinant oligopeptidase B enzymes from Trypanosoma cruzi (Tc-OP) and Trypanosoma brucei (Tb-OP), evaluating the specificity of their S3, S2, S1', S2' and S3' subsites. Five series of internally quenched fluorescent peptides based on the substrate Abz-AGGRGAQ-EDDnp [where Abz is o -aminobenzoic acid and EDDnp is N -(2,4 dinitrophenyl)ethylenediamine] were designed to contain amino acid residues with side chains of a minimum size, and each residue position of this substrate was modified. Synthetic peptides of different lengths derived from the human kininogen sequence were also examined, and peptides of up to 17 amino acids were found to be hydrolysed by Tc-OP and Tb-OP. These two oligopeptidases were essentially arginyl hydrolases, since for all peptides examined the only cleavage site was the Arg-Xaa bond. We also demonstrated that Tc-OP and Tb-OP have a very specific carboxypeptidase activity for basic amino acids, which depends on the presence of at least of a pair of basic amino acids at the C-terminal end of the substrate. The peptide with triple Arg residues (Abz-AGRRRAQ-EDDnp) was an efficient substrate for Tc-OP and Tb-OP: the Arg-Ala peptide bond was cleaved first and then two C-terminal Arg residues were successively removed. The S1' subsite seems to be an important determinant of the specificity of both enzymes, showing a preference for Tyr, Ser, Thr and Gln as hydrogen donors. The presence of these amino acids at P1' resulted in substrates that were hydrolysed with K (m) values in the sub-micromolar range. Taken together, this work supports the view that oligopeptidase B is a specialized protein-processing enzyme with a specific carboxypeptidase activity. Excellent substrates were obtained for Tb-OP and Tc-OP (Abz-AMRRTISQ-EDDnp and Abz-AHKRYSHQ-EDDnp respectively), which were hydrolysed with remarkably high k (cat) and low K (m) values. PMID- 12737624 TI - Pigment-epithelium-derived factor (PEDF) occurs at a physiologically relevant concentration in human blood: purification and characterization. AB - Pigment epithelium-derived factor (PEDF) inhibits the formation of blood vessels in the eye by inducing apotosis in actively dividing endothelial cells. The activity of PEDF equals or supersedes that of other anti-angiogenic factors, including angiostatin, endostatin and thrombospondin-1. In addition, PEDF has the potential to promote the survival of neurons and affect their differentiation. Here we show that PEDF is present in plasma at a concentration of approx. 100 nM (5 microg/ml) or twice the level required to inhibit aberrant blood-vessel growth in the eye. Thus the systemic delivery of PEDF has the potential to affect angiogenesis or neurotrophic processes throughout the body, significantly expanding the putative physiological role of the protein. A complete map of all post-translational modifications revealed that authentic plasma PEDF carries an N terminal pyroglutamate blocking group and an N-linked glycan at position Asn266. The pyroglutamate residue may regulate the activity of PEDF analogously to the manner in which it regulates thyrotropin-releasing hormone. PMID- 12737625 TI - Effect of spermine synthase on the sensitivity of cells to anti-tumour agents. AB - The role of spermine in the sensitivity of cells to various established and experimental anti-tumour agents was examined, using paired cell lines that possess or lack spermine synthase. All spermine-synthase-deficient cells had no detectable spermine, and elevated spermidine, content. Spermine content did not alter the cell growth rate. There was little or no difference in sensitivity of immortalized mouse embryonic fibroblasts to doxorubicin, etoposide, cisplatin, methylglyoxal bis(guanylhydrazone) or H(2)O(2) and only a slight increase in sensitivity to vinblastine and nocodazole. However, the absence of spermine clearly increased the sensitivity to 1,3-bis(2-chloroethyl)- N -nitrosourea, suggesting that depletion of spermine may be a useful way to increase the anti neoplastic effects of anti-tumour agents that form chloroethyl-mediated interstrand DNA cross-links. The effects of spermine on the response to polyamine analogues (which have been proposed to be useful anti-neoplastic agents) were complex, and depended on the compound examined and on the cells tested. Sensitivity to CHENSpm ( N (1)-ethyl- N (11)-[(cycloheptyl)methyl]-4,8 diazaundecane) was substantially greater in immortalized fibroblasts that lack spermine. In contrast, BE-3-4-3 [ N (1), N (12)-bis(ethyl)spermine] and BE-3-3-3 [ N (1), N (11)-bis(ethyl)norspermine] were more active against cells that contained spermine. The presence of spermine correlated with a greater induction of spermidine/spermine- N (1)-acetyltransferase by BE-3-3-3, which is consistent with suggestions that this induction is important for the response to this drug. These findings support the concepts that different polyamine analogues have different sites of action and that CHENSpm has a different site of action from BE 3-3-3. PMID- 12737626 TI - Decreased mitochondrial tRNALys steady-state levels and aminoacylation are associated with the pathogenic G8313A mitochondrial DNA mutation. AB - Mutations in human mitochondrial tRNA genes cause a number of multisystemic disorders. A G-to-A transition at position 8313 (G8313A) transition in the mitochondrial tRNALys gene has been associated with a childhood syndrome characterized by gastrointestinal-system involvement and encephaloneuropathy. We have used transmitochondrial cybrid clones harbouring patient-derived mitochondrial DNA with the G8313A mutation for the study of the molecular pathogenesis. Our results showed that mutant mitochondrial cybrids respired poorly, and had severely defective mitochondrial protein synthesis and respiratory-chain-enzyme activity. Mutant cybrids also showed a marked decrease in tRNALys steady-state levels and aminoacylation, suggesting that these molecular abnormalities may underlie the pathogenesis of the mitochondrial G8313A mutation. PMID- 12737627 TI - A novel protein phosphatase 2A (PP2A) is involved in the transformation of human protozoan parasite Trypanosoma cruzi. AB - Here we provide evidence for a critical role of PP2As (protein phosphatase 2As) in the transformation of Trypanosoma cruzi. In axenic medium at pH 5.0, trypomastigotes rapidly transform into amastigotes, a process blocked by okadaic acid, a potent PP2A inhibitor, at concentrations as low as 0.1 microM. 1 Norokadaone, an inactive okadaic acid analogue, did not affect the transformation. Electron microscopy studies indicated that okadaic acid-treated trypomastigotes had not undergone ultrastructural modifications, reinforcing the idea that PP2A inhibits transformation. Using a microcystin-Sepharose affinity column we purified the native T. cruzi PP2A. The enzyme displayed activity against 32P-labelled phosphorylase a that was inhibited in a dose-dependent manner by okadaic acid. The protein was also submitted to MS and, from the peptides obtained, degenerate primers were used to clone a novel T. cruzi PP2A enzyme by PCR. The isolated gene encodes a protein of 303 amino acids, termed TcPP2A, which displayed a high degree of homology (86%) with the catalytic subunit of Trypanosoma brucei PP2A. Northern-blot analysis revealed the presence of a major 2.1-kb mRNA hybridizing in all T. cruzi developmental stages. Southern blot analysis suggested that the TcPP2A gene is present in low copy number in the T. cruzi genome. These results are consistent with the mapping of PP2A genes in two chromosomal bands by pulsed-field gel electrophoresis and chromoblot hybridization. Our studies suggest that in T. cruzi PP2A is important for the complete transformation of trypomastigotes into amastigotes during the life cycle of this protozoan parasite. PMID- 12737628 TI - The acetyltransferase 60 kDa trans-acting regulatory protein of HIV type 1 interacting protein (Tip60) interacts with the translocation E26 transforming specific leukaemia gene (TEL) and functions as a transcriptional co-repressor. AB - The translocation E26 transforming-specific (ETS) leukaemia (TEL), alias the ETS variant (ETV6), gene is expressed in most human tissues and encodes a transcriptional repressor. The TEL gene is involved in more than 40 different chromosomal translocations associated with haematological malignancies. As little is known about the function of intact TEL, we searched for TEL-interacting proteins by yeast two-hybrid screening. Among the interacting partners, we identified the histone acetyltransferase protein Tip60 [60 kDa trans-acting regulatory protein of HIV type 1 (Tat)-interacting protein]. The interaction was reproduced in vitro, and in mammalian cells we mapped the interaction regions in TEL to the ETS domain and those in Tip60 to the MYST ('MOZ, Ybf2/Sas3, SAS2 and Tip60', where MOZ stands for male absent on the first, SAS for something about silencing and Ybf2 for identical with SAS2) region. Detailed analysis of the Tip60 MYST domain by introduction of point mutations revealed that an N-terminal C2HC zinc finger was essential for interaction with TEL. Finally, we showed that Tip60 functions in a reporter system as a co-repressor in TEL-mediated transcription repression. PMID- 12737629 TI - Dictyostelium discoideum protein phosphatase-1 catalytic subunit exhibits distinct biochemical properties. AB - Protein phosphatase-1 (PP1) is expressed ubiquitously and is involved in many eukaryotic cellular functions, although PP1 enzyme activity could not be detected in the social amoeba Dictyostelium discoideum cell extracts. In the present paper, we show that D. discoideum has a single copy gene that codes for the catalytic subunit of PP1 (DdPP1c). DdPP1c is expressed throughout the D. discoideum life cycle with constant levels of mRNA, and its protein and amino acid sequence show a mean identity of 80% with other PP1c enzymes. However, it has a distinctive difference: the substitution of a phenylalanine residue (Phe(269) in the DdPP1c) for a highly conserved cysteine residue (Cys(273) in rabbit PP1c) in a region that was shown to have a critical role in the interaction of rabbit PP1c with toxin inhibitors. Wild-type DdPP1c and an engineered mutant form in which Phe(269) was replaced by a cysteine residue were expressed in Escherichia coli. Both recombinant activities were similarly inhibited by okadaic acid, tautomycin and microcystin. However, the Phe(269)- >Cys mutation resulted in a large increase in enzyme activity towards phosphorylase a and a higher sensitivity to calyculin A. These results, together with the molecular modelling of DdPP1c structure, indicate that the Phe(269) residue, which occurs naturally in D. discoideum, confers distinct biochemical properties on this enzyme. PMID- 12737630 TI - Role of activator protein-1 in the down-regulation of the human CYP2J2 gene in hypoxia. AB - The cytochrome P450 (CYP) 2J2 arachidonic acid epoxygenase gene was down regulated at a pre-translational level in human hepatoma-derived HepG2 cells incubated in a hypoxic environment; under these conditions, the expression of c Jun and c-Fos mRNA and protein was increased. The 5'-upstream region of the CYP2J2 gene was isolated by amplification of a 2341 bp fragment and putative regulatory elements that resembled activator protein-1 (AP-1)-like sequences were identified. From transient transfection analysis, c-Jun was found to strongly activate a CYP2J2 -luciferase reporter construct, but co-transfection with plasmids encoding c-Fos or c-Fos-related antigens, Fra-1 and -2, abrogated reporter activity. Using a series of deletion-reporter constructs, a c-Jun responsive module was identified between bp -152 and -50 in CYP2J2 : this region contained an AP-1-like element between bp -56 and -63. The capacity of this element to interact directly with c-Jun, but not c-Fos, was confirmed by electromobility-shift assay analysis. Mutagenesis of the -56/-63 element abolished most, but not all, of the activation of CYP2J2 by c-Jun, thus implicating an additional site within the c-Jun-responsive region. The present results establish an important role for c-Jun in the control of CYP2J2 expression in liver cells. Activation of c-Fos expression by hypoxia promotes the formation of c-Jun/c-Fos heterodimers, which decrease the binding of c-Jun to the CYP2J2 upstream region, leading to gene down-regulation. PMID- 12737631 TI - Functional characterization of the human phosphodiesterase 7A1 promoter. AB - In this paper, the human phosphodiesterase 7A1 (h PDE7A1 ) promoter region was identified and functionally characterized. Transient transfection experiments indicated that a 2.9 kb fragment of the h PDE7A1 5'-flanking region, to position 2907, has strong promoter activity in Jurkat T-cells. Deletion analysis showed that the proximal region, up to position -988, contains major cis -regulatory elements of the h PDE7A1 promoter. This minimal promoter region contains a regulatory CpG island which is essential for promoter activity. The CpG island contains three potential cAMP-response-element-binding protein (CREB)-binding sites that, as judged by in vivo dimethyl sulphate (DMS) footprinting, are occupied in Jurkat T-cells. Moreover, over-expression of CREB results in increased promoter activity, but, on the other hand, promoter activity decreases when a dominant-negative form of CREB (KCREB) is over-expressed. In vivo DMS footprinting strongly indicates that other transcription factors, such Ets-2, nuclear factor of activated T-cells 1 (NFAT-1) and nuclear factor kappaB (NF kappaB), might also contribute to the regulation of h PDE7A1 promoter. Finally, h PDE7A1 promoter was found to be induced by treatment with PMA, but not by treatment with dibutyryl cAMP or forskolin. These results provide insights into the factors and mechanisms that regulate expression of the h PDE7A gene. PMID- 12737632 TI - A novel function of tissue-type transglutaminase: protein disulphide isomerase. AB - We have found that tissue-type transglutaminase (tTG), also called TGc, TGase2 and Galpha(h), has the activity of protein disulphide isomerase (PDI). We have shown that tTG converts completely reduced/denatured inactive RNase A molecule to the native active enzyme. The PDI activity of tTG was strongly inhibited by bacitracin, which is a frequently used inhibitor of conventional PDI activity. It was substantially inhibited by the simultaneous presence of other potential substrate proteins such as completely reduced BSA, but not by native BSA. This activity was especially high in the presence of GSSG, but not GSH. The addition of GSH to the reaction mixture in the presence of GSSG at a fixed concentration up to at least 200-fold excess did not very substantially inhibit the PDI activity. It is possible that tTG can exert PDI activity in a fairly reducing environment like cytosol, where most of tTG is found. It is quite obvious from the following observations that PDI activity of tTG is catalysed by a domain different from that used for the transglutaminase reaction. Although the alkylation of Cys residues in tTG completely abolished the transglutaminase activity, as was expected, it did not affect the PDI activity at all. This PDI activity did not require the presence of Ca(2+). It was not inhibited by nucleotides including GTP at all, unlike the other activity of tTG. PMID- 12737633 TI - Potent P2Y6 receptor mediated contractions in human cerebral arteries. AB - BACKGROUND: Extracellular nucleotides play an important role in the regulation of vascular tone and may be involved in cerebral vasospasm after subarachnoidal haemorrhage. This study was designed to characterise the contractile P2 receptors in endothelium-denuded human cerebral and omental arteries. The isometric tension of isolated vessel segments was recorded in vitro. P2 receptor mRNA expression was examined by RT-PCR. RESULTS: In human cerebral arteries, the selective P2Y6 receptor agonist, UDPbetaS was the most potent of all the agonists tested (pEC50 = 6.8 PlusMinus; 0.7). The agonist potency; UDPbetaS > alphabeta-MeATP > UTPgammaS > ATPgammaS > ADPbetaS = 0, indicated the presence of contractile P2X1 P2Y2, P2Y4 and P2Y6, but not P2Y1 receptors, in human cerebral arteries. In human omental arteries, UDPbetaS was inactive. The agonist potency; alphabeta-MeATP > ATPgammaS = UTPgammaS > ADPbetaS = UDPbetaS = 0, indicated the presence of contractile P2X1, and P2Y2 receptors, but not P2Y1 or P2Y6 receptors, in human omental arteries. RT-PCR analysis of endothelium-denuded human cerebral and omental arteries demonstrated P2X1, P2Y1, P2Y2 and P2Y6 receptor mRNA expression. There were no bands for the P2Y4 receptor mRNA in the omental arteries, while barely detectable in the cerebral arteries. CONCLUSIONS: P2Y6 receptors play a prominent role in mediating contraction of human cerebral arteries. Conversely, no such effect can be observed in human omental arteries and previous results confirm the absence of P2Y6 receptors in human coronary arteries. The P2Y6 receptor might be a suitable target for the treatment of cerebral vasospasm. PMID- 12737635 TI - What is the relationship between the minimally important difference and health state utility values? The case of the SF-6D. AB - BACKGROUND: The SF-6D is a new single summary preference-based measure of health derived from the SF-36. Empirical work is required to determine what is the smallest change in SF-6D scores that can be regarded as important and meaningful for health professionals, patients and other stakeholders. OBJECTIVES: To use anchor-based methods to determine the minimally important difference (MID) for the SF-6D for various datasets. METHODS: All responders to the original SF-36 questionnaire can be assigned an SF-6D score provided the 11 items used in the SF 6D have been completed. The SF-6D can be regarded as a continuous outcome scored on a 0.29 to 1.00 scale, with 1.00 indicating "full health". Anchor-based methods examine the relationship between an health-related quality of life (HRQoL) measure and an independent measure (or anchor) to elucidate the meaning of a particular degree of change. One anchor-based approach uses an estimate of the MID, the difference in the QoL scale corresponding to a self-reported small but important change on a global scale. Patients were followed for a period of time, then asked, using question 2 of the SF-36 as our global rating scale, (which is not part of the SF-6D), if there general health is much better (5), somewhat better (4), stayed the same (3), somewhat worse (2) or much worse (1) compared to the last time they were assessed. We considered patients whose global rating score was 4 or 2 as having experienced some change equivalent to the MID. In patients who reported a worsening of health (global change of 1 or 2) the sign of the change in the SF-6D score was reversed (i.e. multiplied by minus one). The MID was then taken as the mean change on the SF-6D scale of the patients who scored (2 or 4). RESULTS: This paper describes the MID for the SF-6D from seven longitudinal studies that had previously used the SF-36. CONCLUSIONS: From the seven reviewed studies (with nine patient groups) the MID for the SF-6D ranged from 0.010 to 0.048, with a weighted mean estimate of 0.033 (95% CI: 0.029 to 0.037). The corresponding Standardised Response Means (SRMs) ranged from 0.11 to 0.48, with a mean of 0.30 and were mainly in the "small to moderate" range using Cohen's criteria, supporting the MID results. Using the half-standard deviation (of change) approach the mean effect size was 0.051 (range 0.033 to 0.066). Further empirical work is required to see whether or not this holds true for other patient groups and populations. PMID- 12737634 TI - Isolation and characterization of gelatin-binding proteins from goat seminal plasma. AB - A family of proteins designated BSP-A1, BSP-A2, BSP-A3 and BSP-30 kDa (collectively called BSP proteins for Bovine Seminal Plasma proteins) constitute the major protein fraction in the bull seminal plasma. These proteins interact with choline phospholipids on the sperm surface and play a role in the membrane stabilization (decapacitation) and destabilization (capacitation) process. Homologous proteins have been isolated from boar and stallion seminal plasma. In the current study we report the isolation and preliminary characterization of homologous proteins from goat seminal plasma. Frozen semen (-80 degrees C) was thawed and centrifuged to remove sperm. The proteins in the supernatant were precipitated by the addition of cold ethanol. The precipitates were dissolved in ammonium bicarbonate and lyophilised. The lyophilised proteins were dissolved in phosphate buffer and loaded onto a gelatin-agarose column, which was previously equilibrated with the same buffer. The column was successively washed with phosphate buffer, with phosphate buffer saline and with 0.5 M urea in phosphate buffer saline to remove unadsorbed proteins, and the adsorbed proteins were eluted with 5 M urea in phosphate buffer saline. Analysis of pooled, dialysed and lyophilised gelatin-agarose adsorbed protein fraction by SDS-PAGE indicated the presence of four protein bands that were designated GSP-14 kDa, GSP-15 kDa, GSP 20 kDa and GSP-22 kDa (GSP, Goat Seminal Plasma proteins). Heparin-affinity chromatography was then used for the separation of GSP-20 and -22 kDa from GSP-14 and -15 kDa. Finally, HPLC separation permitted further isolation of each one from the other. Amino acid sequence analysis of these proteins indicated that they are homologous to BSP proteins. In addition, these BSP homologs bind to hen's egg-yolk low-density lipoproteins. These results together with our previous data indicate that BSP family proteins are ubiquitous in mammalian seminal plasma, exist in several forms in each species and possibly play a common biological role. PMID- 12737637 TI - HIV/AIDS salvage therapy. Interview by Vicki Glaser. PMID- 12737636 TI - Sympatric Plasmodium falciparum isolates from Venezuela have structured var gene repertoires. AB - BACKGROUND: The human malaria parasite Plasmodium falciparum expresses adhesins belonging to the erythrocyte membrane protein 1 (PfEMP1) family on the surface of the infected host erythrocyte. These antigens elicit a strain-specific antibody response that is associated with protection from disease. During clonal expansion of blood-stage parasites, the surface phenotype of the infected erythrocyte changes because of transcriptional switching among the 40 to 50 members of the highly polymorphic var multi-gene family which encode PfEMP1 variants. Studies to date have compared var repertoires of natural isolates from various geographical locations but have not addressed any within-population structure that may exist among repertoires. METHODS: Distinct parasite genotypes from a single population co-circulating among a defined group of hosts were selected. PCR products encoding the DBL-alpha domain of PfEMP-1 were cloned and sequenced from each of three isolates. Repertoire similarity was statistically evaluated using combinatorial analysis. The chromosomal location of shared sequences was inferred from similarity to dbl-alpha of known location in the 3D7 genome. RESULTS: Sympatric parasites were found to share few var gene sequences, even when alleles at other polymorphic loci were shared. A number of the sequences shared by at least two of the isolates studied were found to be related to 3D7 genomic sequences with non-telomeric chromosomal locations, or atypical domain structures, which may represent globally conserved loci. CONCLUSION: The parasite population studied is structured, with minimal overlap in PfEMP1 repertoires. The var gene family accumulates diversity more rapidly than other antigen genes examined. This may be facilitated by ectopic recombination among the sub telomeric regions of P. falciparum chromosomes. PMID- 12737638 TI - Elevated serum levels of soluble immune activation markers are associated with increased risk for death in HAART-naive HIV-1-infected patients. AB - The aim of this study was to determine the serum levels of soluble markers reflecting different aspects of immune activation in HIV-1-infected patients, and assess their prognostic significance for occurrence of AIDS-related death before the advent of the highly active antiretroviral treatment (HAART). Serum concentrations of the soluble forms of interleukin-2 receptors (sIL-2R), intercellular adhesion molecule-1 (sICAM-1), and E-selectin (sEs) have been determined in a cohort of 64 HIV-1-infected patients, between 1990-1993. The patients were followed prospectively with regular visits at the outpatient department. Follow-up time was censored at January 1, 1997, the date after which HAART was introduced. The median follow-up time was 46 months (range, 2-78 months). By the end of follow-up, 34 subjects had died. Baseline levels of all three soluble markers were significantly lower in subjects who remained alive during the follow-up compared to subjects who died. Univariate analysis showed that individual sIL-2R and sICAM-1, but not sEs measurements, were significantly associated with time to death (p = 0.008 and 0.003, respectively). Even after adjustment for age and CD4+ T-cell counts sIL-2R measurements remained significantly prognostic. Sensitivity analysis using follow-up time to year 2000 confirmed these results. Our data suggest that assessment of the immune activation status using the easily measured levels of circulating markers may provide additional information about the risk of AIDS-related death. Further studies are needed to assess the effect of HAART on the levels of immune activation markers and their prognostic value. PMID- 12737639 TI - Use of complementary and alternative medicine by HIV-infected outpatients in Ontario, Canada. AB - Little is known about complementary and alternative medicine (CAM) use in Canadian patients with HIV. We sought to determine the prevalence of CAM use by patients attending HIV outpatient clinics in Ontario, Canada, and to compare the characteristics of users and nonusers. Impact of CAM definition on reported utilization rates was also assessed, specifically in relation to the inclusion and exclusion of vitamins, minerals, and multivitamins in CAM definition. In person interviews were conducted between 1999 and 2001 with 104 HIV-positive outpatients enrolled in the HIV Ontario Observational Database project (HOOD) and attending HIV outpatient clinics in Ontario. Self-reported CAM utilization and demographic data were collected. Clinical data were obtained from medical chart review. Seventy-seven percent of participants reported current CAM use. Inclusion of vitamins and minerals (CAMVIT) increased this estimate to 89%. Nearly all patients used CAM in conjunction with antiretroviral medications. Out of pocket costs ranged from CAD$0 to more than CAD$250 per month. Most patients reported CAM use was beneficial and had improved their overall health. Female gender, HIV risk group, number of prescriptions, and overall number of drugs used were associated with CAM use. CAM use in Canadian patients with HIV is extremely common, with higher use among women. The definition of CAM has a substantial impact both on reported prevalence rates and on predictors of CAM use. PMID- 12737640 TI - Adherence to HAART regimens. AB - The problem of inadequate adherence to prescribed highly active antiretroviral therapy (HAART) drug regimens to treat HIV infection and AIDS is ubiquitous. Adherence can be inadequate despite both provider and patient understanding of the consequences of nonadherence. Successful long-term treatment of HIV/AIDS requires at least 95% adherence to HAART in order to prevent emergence of drug resistant HIV variants that lead to regimen failure and limit options for future therapy. Despite the prevalence of inadequate adherence, many patients succeed, and HAART has transformed HIV infection into a chronic illness increasingly managed in primary care. The barriers to adherence observed in HIV treatment resemble barriers to the successful treatment of other chronic diseases: regimen complexity, side effects resulting in poor tolerability, patient lifestyle factors, and patient-provider relationships. Treatment of HIV infection has shown that patient-provider collaboration can result in the selection of a lifestyle tailored regimen characterized by convenient dosing, low pill burden, and tolerable side effects that enhances adherence, effectiveness, and the patient's willingness to remain on anti-HIV therapy long term. This review focuses on the current understanding of adherence reporting, improvement of adherence, and, hence, improvement of treatment outcomes in HIV infection and AIDS. PMID- 12737641 TI - Adherence to HIV antiretrovirals among persons with serious mental illness. AB - Despite the absence of empirical evidence, serious mental illness is assumed to be a high risk factor for nonadherence to HIV antiretroviral regimens. To assess antiretroviral adherence among persons with serious mental illness, we conducted a study in which adherence was observed over a 2-week period with electronic monitoring bottle caps and self-report. Forty-seven participants enrolled, with all but two (96%) completing the study. Psychiatric diagnoses included bipolar depression (n = 24), schizophrenia (n = 12), schizoaffective disorder (n = 5), and major depression with psychotic features (n = 6). Mean adherence (proportion of prescribed doses taken) was 66% (standard deviation [SD] = 34), as measured by electronic monitoring; 40% demonstrated at least 90% adherence, but 31% had less than 50% adherence. Self-reported adherence to psychotropics was moderately correlated with self-reported (r = 0.45, p < 0.05) and electronically monitored (r = 0.39, p < 0.05) antiretroviral adherence. Viral load (log(10)) was negatively correlated with electronically monitored (r = -0.28, p < 0.10) and self-reported (r = -0.39, p < 0.05) antiretroviral adherence, after controlling for the length of time on treatment. These findings suggest that many patients with serious mental illness are able to adhere very well to antiretroviral regimens, yet a substantial proportion of our sample displayed poor adherence, indicating the need for research to further assess the factors that influence adherence to antiretrovirals in this population. PMID- 12737643 TI - NEAT results released. PMID- 12737642 TI - Functional status and overall quality of life in a multiethnic HIV-positive population. AB - The objective of this study was to examine the sociodemographic and behavioral variables associated with quality of life (QOL) among multiethnic, economically disadvantaged patients with HIV/AIDS. A cross-sectional survey was conducted in a large inner-city HIV/AIDS clinic serving medically indigent residents of Houston, Texas, and the surrounding area. On arrival at the clinic, patients were systematically approached and asked to complete a questionnaire, offered in both English and Spanish. Demographic characteristics, stage of disease, and behavioral variables were assessed in addition to work-role functioning and overall health-related QOL. Multiple regression analysis was conducted to assess the relationships between the variables of interest. Of 617 patients approached and asked to participate in the study, 385 (62.4%) consented to complete the questionnaire. Demographic composition of the sample was as follows: 78% male, 25% white, 44% black, and 29% Hispanic. Forty-five percent of the participants were infected through men having sex with men (MSM), 35% through heterosexual contact, and 11% through injection drug use. Racial/ethnic minorities reported significantly (p < 0.05) poorer physical functioning and work-role functioning. Participants with higher nadir CD4 cell counts (> 500 per cubic millimeter), MSM HIV exposure, and more education (beyond high school) reported better physical functioning and work-role functioning. Overall QOL and work-related functioning were significantly impaired in this patient population. The impaired functional status findings hold for every social, demographic, and behavioral subgroup examined. Findings suggest that the influence of social inequality persist even among a universally disadvantaged population. PMID- 12737644 TI - Resistance data also presented. PMID- 12737645 TI - Nonendoscopic Nd-YAG 1064 nm PLDN in the treatment of thoracic discogenic pain syndromes. AB - OBJECTIVE: The purpose of the present study was to discover new minimal invasive treatments of discogenic thoracic pain caused by protrusions or extrusions using the promising method of nonendoscopic Nd-YAG 1064 nm PLDN in the lumbar and cervical regions. Because early symptoms of chronic thoracic discogenic pain syndromes have not been characterized, interventional therapy is usually started late and involves a high complication rate. MATERIALS AND METHODS: A prospective controlled clinical study was undertaken by neurologists using Nd-YAG 1064 nm PLDN to treat 42 patients with thoracic disc protrusions and extrusions. Patients with discogenic pain syndromes and MRI-confirmed disc pathology with spinal canal impairment were enrolled; 68 discs were treated. Maximal Nd-YAG laser 1064 nm dose was 1,000 watts per segment. Disc puncture was performed by dorsolateral approach. Monitored parameters were VAS, McNab score, subjective condition, neurological findings and peripheral EMG. A different, independent neurologist examined each case before and after surgery. RESULTS: At 6 weeks after treatment, 41 patients had a successful outcome; only one with a clinical suspicion of spondylodiscitis was dissatisfied. In all others, clinical parameters improved. EMG leaks had disappeared. Combined spastic paresis improved in 2/4 cases. Complications were one pneumothorax, one pleuritis and one suspected spondylodiscitis. CONCLUSION: Pain relief and decompression of spinal structures is effective and immediate by disc vaporization, shrinkage, nociceptor destruction and discogenic kinius denaturation. Nonendoscopic percutaneous Nd-YAG 1064 nm PLDN is a highly effective method for the treatment of thoracic disc disorders with minimally invasive access and is recommended prior to any open surgery. PMID- 12737646 TI - Effect of NASA light-emitting diode irradiation on molecular changes for wound healing in diabetic mice. AB - OBJECTIVE: The purpose of this study was to assess the changes in gene expression of near-infrared light therapy in a model of impaired wound healing. BACKGROUND DATA: Light-Emitting Diodes (LED), originally developed for NASA plant growth experiments in space, show promise for delivering light deep into tissues of the body to promote wound healing and human tissue growth. In this paper we present the effects of LED treatment on wounds in a genetically diabetic mouse model. MATERIALS AND METHODS: Polyvinyl acetal (PVA) sponges were subcutaneously implanted in the dorsum of BKS.Cg-m +/+ Lepr(db) mice. LED treatments were given once daily, and at the sacrifice day, the sponges, incision line and skin over the sponges were harvested and used for RNA extraction. The RNA was subsequently analyzed by cDNA array. RESULTS: Our studies have revealed certain tissue regenerating genes that were significantly upregulated upon LED treatment when compared to the untreated sample. Integrins, laminin, gap junction proteins, and kinesin superfamily motor proteins are some of the genes involved during regeneration process. These are some of the genes that were identified upon gene array experiments with RNA isolated from sponges from the wound site in mouse with LED treatment. CONCLUSION: We believe that the use of NASA light-emitting diodes (LED) for light therapy will greatly enhance the natural wound healing process, and more quickly return the patient to a preinjury/illness level of activity. This work is supported and managed through the Defense Advanced Research Projects Agency (DARPA) and NASA Marshall Space Flight Center-SBIR Program. PMID- 12737647 TI - Interstitial laser hyperthermia and the biological characteristics of tumor: study in a murine model of colorectal liver metastases. AB - BACKGROUND DATA: Percutaneously applied interstitial laser hyperthermia (ILH) is a minimally invasive therapy that is currently used in the treatment of liver metastases. Despite its documented efficacy, theoretical considerations and evidence based on animal studies suggest the potential for stimulating tumor growth, especially following surgery. This study investigates the influence of ILH on tumor behaviour in an animal model of colorectal liver metastases. MATERIALS AND METHODS: A model of colorectal cancer liver metastases in male inbred CBA mice was used. Laser hyperthermia was applied to tumor tissue using a bare optical quartz from a Medilas fibertom 4100 Nd:YAG surgical laser generator. Liver injury by ILH was initially produced in three experimental groups of animals at different time points in the development of metastases. ILH was applied (i) to normal liver 10 days prior to tumor induction, (ii) immediately prior to tumor induction, and (iii) 15 days after tumor induction to achieve approximately 8% liver destruction. Animals were killed 21 days after tumor induction, and the effects of ILH on overall tumor development were compared with controls using stereological assessment of tumor volume and by histology. In a separate experimental group, the effects of ILH on fully established tumors were examined. Suitable tumors were selected 21 days after induction and partially destroyed by ILH at a standard energy setting. Animals were then killed 15 days later, and the growth rate of the residual viable tumors was compared to control tumors having undergone sham procedures. RESULTS: No significant stimulation of tumor growth was evident in any of the experimental groups following ILH, irrespective of the time of application. Incomplete tumor destruction also had no influence on subsequent tumor growth. CONCLUSION: ILH does not influence the biological characteristics of tumors during any stage of the metastatic process. PMID- 12737648 TI - Preparation of root canal orifices by Er:YAG laser irradiation: in vitro and clinical observations. AB - OBJECTIVE: The purpose of the present study was to evaluate the effectiveness of Er:YAG laser irradiation for preparation of root canal orifices in extracted human teeth and several clinical cases. BACKGROUND DATA: Few studies with sufficient data have been conducted in this area. MATERIALS AND METHODS: Orifices of 42 extracted human teeth were prepared by conventional methods using a Peeso reamer or Er:YAG laser device at 250 mJ/pulse and 8 Hz. In the clinical study, the orifices of 11 teeth from 11 patients with irreversible pulpitis were prepared by Er:YAG laser irradiation at 160 mJ/pulse and 8 Hz. Teeth were carefully irradiated using non-contact methods. Evaluation was performed by visual inspection, stereoscopy, radiography, and scanning electron microscopy (SEM). In addition, the efficiency of using the Er:YAG laser was evaluated. RESULTS: In the in vitro study, canal orifices were clearly exposed by laser irradiation in all specimens. In 31 of 36 teeth (86%), orifices were successfully prepared without ledge formation or perforation. SEM observations revealed that irradiated surfaces were slightly rough and scaly, but essentially free from debris and smear layer. In the clinical study, orifices were successfully prepared in 10 of 11 teeth (91%), and no ledge formation or perforation was observed. CONCLUSION: These results suggest that the preparation of root canal orifices by Er:YAG laser irradiation may be useful in most cases, if appropriate parameters are selected. PMID- 12737649 TI - Endoscope-assisted KTP laser sinus clear-out procedure for recurrent ethmoid polyposis. AB - BACKGROUND DATA AND OBJECTIVE: Surgery for recurrent sinusitis with diffuse polyposis is challenging since tortuous sinus anatomy and bleeding tendency can make revision surgery particularly difficult. We report the efficacy in using KTP laser under the assistance of endoscope to perform sinus clear-out procedure for patients with recurrent ethmoid polyposis. MATERIALS AND METHODS: Thirteen patients with recurrent ethmoid polyposis underwent this procedure between November 1996 and July 2000. All were available for follow-up. Patients all had a KTP sinus clear-out procedure (15-20 W continuous mode, spot size 0.4-0.8 mm, contact or near-contact approaches) assisted by endoscope. Sinus endoscopic findings, CAT scan data, operation records and perioperative courses were retrospectively reviewed. The mean follow-up period was 24.7 +/- 11 months. RESULTS: Surgeries all went smoothly with minimal bleeding incurred. All patients demonstrated significant improvement in their sinonasal symptoms. Two cases (15.4%) had localized recurrent polyps. CONCLUSION: With its excellent capabilities in hemostasis, great intra-nasal maneuverability, and flexible operation modes, KTP laser is an ideal alternative to manage recurrent ethmoid polyposis for selected cases. PMID- 12737651 TI - Influence of the frequency of Er:YAG laser on the bond strength of dental enamel. AB - OBJECTIVE: The present study had the aim of evaluating the influence of different frequencies of the Er:YAG laser on adhesive resistance of enamel and one restorative system. BACKGROUND DATA: There have been no reports of studies assessing the influence of the pulse frequency variation of the Er:YAG laser on adhesive resistance of the enamel/resin interface. MATERIALS AND METHODS: Fifty surfaces of enamel from extracted human third molars were planed and divided into five groups at random. Enamel surface treatment was realized by the Er:YAG laser at 80-mJ power and 1-, 2-, 3-, and 4-Hz pulse frequencies, followed by etching. For the control group, only acid conditioning with 37% phosphoric acid for 15 sec was used. The Single Bond/Filtek Z250 system was chosen for the fabrication of the specimens, which were stored in 100% relative humidity for 24 h, at 37 degrees C. The specimens were submitted to tensile resistance tests using a Universal Testing Machine (50 Kgf and 0.5 mm/min). RESULTS: The mean values in MPa were 1 Hz, 25.58 (+/-6.16); 2 Hz, 25.58 (+/-3.79); 3 Hz, 21.34 (+/-3.78); 4 Hz, 21.17 (+/-3.13); and phosphoric acid only, 22.44 (+/-7.0). Data were submitted to statistical analysis using ANOVA, and there was no significant difference in tensile resistance between the studied groups. CONCLUSION: The results suggest that the Er:YAG laser, with 80-mJ power associated with acid conditioning at 1-, 2-, 3-, and 4-Hz frequencies, did not present significant improvement in tensile bonding of enamel, as compared to acid conditioning only. PMID- 12737650 TI - He-Ne laser on microcrystalline arthropathies. AB - OBJECTIVE: The objective of this work is to assess the anti-inflammatory capacity of He-Ne laser therapy as determined by the plasmatic levels of inflammatory markers, fibrinogen, and TNFalpha and by histopathological study in rats with arthropathy induced by calcium pyrophosphate crystals. BACKGROUND DATA: Microcrystalline arthropathies are a group of diseases characterized by the deposit of different crystals in joints. MATERIALS AND METHODS: Two milligrams of dicalcium pyrophosphate crystals (DCPP) were injected in both joints of the lower limbs of rats during 2 days. A group was treated with laser of He-Ne (6 mW) on the injected joints during 3 consecutive days. After 96 h of the first injection, animals were sacrificed to determine TNFalpha using the ELISA method and fibrinogen was assessed using spectrophotometry. Sections from the lower limbs were used for histopathology. RESULTS: A statistically significant increase (p < 0.001) in plasma fibrinogen levels and TNFalpha was noted between the control group and the laser-treated group. The histological transversal section of a posterior limb joint of a rat injected with DCPP showed fibroadipose tissue with diffuse chronic infiltrate. The histopathology of the group of rats injected with DCPP and subsequently treated with He-Ne laser showed no inflammatory response. CONCLUSION: He-Ne laser treatment in the microcrystalline arthropathy induced in rats by DCPP injection might have an antiinflammatory effect, evaluated by fibrinogen plasma levels and TNF-alpha (inflammatory markers) and by the histopathology regressive process. PMID- 12737653 TI - Laser literature watch. PMID- 12737654 TI - Mechanisms underlying the impacts of exotic plant invasions. AB - Although the impacts of exotic plant invasions on community structure and ecosystem processes are well appreciated, the pathways or mechanisms that underlie these impacts are poorly understood. Better exploration of these processes is essential to understanding why exotic plants impact only certain systems, and why only some invaders have large impacts. Here, we review over 150 studies to evaluate the mechanisms underlying the impacts of exotic plant invasions on plant and animal community structure, nutrient cycling, hydrology and fire regimes. We find that, while numerous studies have examined the impacts of invasions on plant diversity and composition, less than 5% test whether these effects arise through competition, allelopathy, alteration of ecosystem variables or other processes. Nonetheless, competition was often hypothesized, and nearly all studies competing native and alien plants against each other found strong competitive effects of exotic species. In contrast to studies of the impacts on plant community structure and higher trophic levels, research examining impacts on nitrogen cycling, hydrology and fire regimes is generally highly mechanistic, often motivated by specific invader traits. We encourage future studies that link impacts on community structure to ecosystem processes, and relate the controls over invasibility to the controls over impact. PMID- 12737655 TI - A one-step organelle capture: gynogenetic kiwifruits with paternal chloroplasts. AB - Androgenesis, the development of a haploid embryo from a male nucleus, has been shown to result in the instantaneous uncoupling of the transmission of the organelle and nuclear genomes (with the nuclear genome originating from the male parent only and the organelle genomes from the female parent). We report, for the first time, uncoupling resulting from gynogenesis, in Actinidia deliciosa (kiwifruit), a plant species known for its paternal mode of chloroplast inheritance. After pollen irradiation, transmission of nuclear genes from the pollen parent to the progeny was inhibited, but transmission of the chloroplast genome was not. This demonstrates that plastids can be discharged from the pollen tube into the egg with little or no concomitant transmission of paternal nuclear genes. Such events of opposite inheritance of the organelle and nuclear genomes must be very rare in nature and are unlikely to endanger the long-term stability of the association between the different genomes of the cell. However, they could lead to incongruences between organelle gene trees and species trees and may constitute an alternative to the hybridization/introgression scenario commonly invoked to account for such incongruences. PMID- 12737656 TI - Initial frequency of alleles conferring resistance to Bacillus thuringiensis poplar in a field population of Chrysomela tremulae. AB - Globally, the estimated total area planted with transgenic plants producing Bacillus thuringiensis (Bt) toxins was 12 million hectares in 2001. The risk of target pests becoming resistant to these toxins has led to the implementation of resistance-management strategies. The efficiency and sustainability of these strategies, including the high-dose plus refuge strategy currently recommended for North American maize, depend on the initial frequency of resistance alleles. In this study, we estimated the initial frequencies of alleles conferring resistance to transgenic Bt poplars producing Cry3A in a natural population of the poplar pest Chrysomela tremulae (Coleoptera: Chrysomelidae). We used the F(2) screen method developed for detecting resistance alleles in natural pest populations. At least three parents of the 270 lines tested were heterozygous for a major Bt resistance allele. We estimated mean resistance-allele frequency for the period 1999-2001 at 0.0037 (95% confidence interval = 0.00045-0.0080) with a detection probability of 90%. These results demonstrate that (i) the F(2) screen method can be used to detect major alleles conferring resistance to Bt-producing plants in insects and (ii) the initial frequency of alleles conferring resistance to Bt toxin can be close to the highest theoretical values that are expected prior to the use of Bt plants if considering fitness costs and typical mutation rates. PMID- 12737657 TI - Synergistic effects of food and predators on annual reproductive success in song sparrows. AB - The behaviour literature is full of studies showing that animals in every taxon balance the probability of acquiring food with the risk of being preyed upon. While interactions between food and predators clearly operate at an individual scale, population-scale studies have tended to focus on only one factor at a time. Consequently, interactive (or 'synergistic') effects of food and predators on whole populations have only twice before been experimentally demonstrated in mammals. We conducted a 2 x 2 experiment to examine the joint effects of food supply and predator pressure on the annual reproductive success of song sparrows (Melospiza melodia). Our results show that these two factors do not operate in an additive way, but instead have a synergistic effect on reproduction. Relative to controls, sparrows reared 1.1 more young when food was added and 1.3 more when predator pressure was low. When these treatments were combined 4.0 extra young were produced, almost twice as many as expected from an additive model. These results are a cause for optimism for avian conservation because they demonstrate that remedial actions, aimed at simultaneously augmenting food and reducing predators, can produce dramatic increases in reproductive success. PMID- 12737658 TI - Genetic evidence for intra- and interspecific slavery in honey ants (genus Myrmecocystus). AB - The New World honey ant species Myrmecocystus mimicus is well known for its highly stereotyped territorial tournaments, and for the raids on conspecific nests that can lead to intraspecific slavery. Our results from mitochondrial and nuclear markers show that the raided brood emerges in the raiding colony and is subsequently incorporated into the colony's worker force. We also found enslaved conspecifics in a second honey ant species, M. depilis, the sister taxon of M. mimicus, which occurs in sympatry with M. mimicus at the study site. Colonies of this species furthermore contained raided M. mimicus workers. Both species have an effective mating frequency that is not significantly different from 1. This study provides genetic evidence for facultative intra- and interspecific slavery in the genus Myrmecocystus. Slavery in ants has evolved repeatedly and supposedly by different means. We propose that, in honey ants, secondary contact between two closely related species that both exhibit intraspecific slavery gave rise to an early form of facultative interspecific slavery. PMID- 12737659 TI - Proximate and ultimate control of sex ratios in Myrmica brevispinosa colonies. AB - The literature on sex ratio evolution in ant colonies is dominated by inclusive fitness arguments. In general, genetic theory makes good predictions about sexual investment in ant populations, but understanding colony-level variance in sex investment ratios has proven more difficult. Recently, however, more studies have addressed ecological factors that influence colony-level sex investment ratios. Food availability, in particular, has been manipulated because larval nutrition influences female caste determination, thus implying that resource availability should be of critical importance for colony sex investment ratios. However, results from food supplementation experiments are equivocal, and it is clear that ant colony response to food supplementation is dependent on the ecological background of the population. We presented field colonies of the ant Myrmica brevispinosa with two food types (proteins and carbohydrates), and assessed their relative impact on colony-level sex investment ratios. We show that colonies receiving carbohydrate enhancement invested in more female sexuals and produced more female-biased sex allocation ratios than protein-fed or control colonies. Thus, our study is the first, to our knowledge, to demonstrate that sex ratios in social insect colonies might be sensitive to resource quality. Male investment was not influenced by food treatment, but was positively correlated with colony size. Therefore, the shift in sex ratios in our study must have been mediated through nutritional influences on female caste determination rather than male brood elimination. We also used our data to evaluate evidence for sex ratio compensation by queenright colonies in response to male production by workers from orphaned colonies. PMID- 12737660 TI - Does morality have a biological basis? An empirical test of the factors governing moral sentiments relating to incest. AB - Kin-recognition systems have been hypothesized to exist in humans, and adaptively to regulate altruism and incest avoidance among close genetic kin. This latter function allows the architecture of the kin recognition system to be mapped by quantitatively matching individual variation in opposition to incest to individual variation in developmental parameters, such as family structure and co residence patterns. Methodological difficulties that appear when subjects are asked to disclose incestuous inclinations can be circumvented by measuring their opposition to incest in third parties, i.e. morality. This method allows a direct test of Westermarck's original hypothesis that childhood co-residence with an opposite-sex individual predicts the strength of moral sentiments regarding third party sibling incest. Results support Westermarck's hypothesis and the model of kin recognition that it implies. Co-residence duration objectively predicts genetic relatedness, making it a reliable cue to kinship. Co-residence duration predicts the strength of opposition to incest, even after controlling for relatedness and even when co-residing individuals are genetically unrelated. This undercuts kin-recognition models requiring matching to self (through, for example, major histocompatibility complex or phenotypic markers). Subjects' beliefs about relatedness had no effect after controlling for co-residence, indicating that systems regulating kin-relevant behaviours are non-conscious, and calibrated by co-residence, not belief. PMID- 12737661 TI - No slave to sex. AB - Fully asexual lineages cannot purge accumulating mutations from their genome through recombination. In ancient asexuals that have persisted without sex for millions of years, this should lead to high allelic divergences (the 'Meselson effect') as has been shown for bdelloid rotifers. Homogenizing mechanisms can counter this effect, resulting in low genetic diversity within and between individuals. Here, we show that the ancient asexual ostracod species Darwinula stevensoni has very low nucleotide sequence divergence in three nuclear regions. Differences in genetic diversity between embryos and adults furthermore indicate that up to half of the observed genetic changes in adults can be caused by somatic mutations. Likelihood permutation tests confirm the presence of gene conversion in the multi-copy internal transcribed spacer sequence, but reject rare or cryptic forms of sex as a general explanation for the low genetic diversity in D. stevensoni. Other special mechanisms (such as highly efficient DNA repair) might have been selected for in this ancient asexual to overcome the mutational load and Muller's ratchet. In this case, our data support these hypotheses on the prevalence of sex, even if the two extant ancient asexual groups (bdelloids and darwinulids) seem to follow opposite evolutionary strategies. PMID- 12737662 TI - Specialized cheating of the ectomycorrhizal symbiosis by an epiparasitic liverwort. AB - Many non-photosynthetic vascular plants in 10 diverse families obtain all of their carbon from fungi, but in most cases the fungi and the ultimate sources of carbon are unknown. In a few cases, such plants have been shown to be epiparasitic because they obtain carbon from neighbouring green plants through shared mycorrhizal fungi. In all such cases, the epiparasitic plants have been found to specialize upon narrow lineages of ecto- or arbuscular mycorrhizal fungi. Here we show that a non-vascular plant, the non-photosynthetic liverwort Cryptothallus mirabilis, is epiparasitic and is specialized on Tulasnella species that form ectomycorrhizae with surrounding trees at four locations in England, France and Portugal. By using microcosm experiments we show that the interaction with Tulasnella is necessary for growth of Cryptothallus, and by using labelling experiments we show that (14)CO(2) provided to birch seedlings is transferred to Cryptothallus by Tulasnella. This is one of the first documented cases of epiparasitism by a non-vascular plant and of ectomycorrhizal formation by Tulasnella. These results broaden the emerging association between epiparasitism and mycorrhizal specialization into a new class of plants and a new order of fungi. PMID- 12737663 TI - The influence of cellular physiology on the initiation of mutational pathways in Escherichia coli populations. AB - The factors affecting the direction of evolutionary pathways and the reproducibility of adaptive responses were investigated under closely related but non-identical conditions. Replicate chemostat cultures of Escherichia coli were compared when adapting to partial or severe glucose limitation. Four independent populations used a reproducible sequence of early mutational changes under both conditions, with rpoS mutations always occurring first before mgl. However, there were interesting differences in the timing of mutational sweeps: rpoS mutations appeared in a clock-like fashion under both partial and severe glucose limitation, while mgl sweeps arose under both conditions but at different times. Interestingly, malT and mlc mutations appeared only under severe limitation. Even though the ancestors were genotypically identical, the semi-differentiated properties of bacteria growing with mild or severe glucose limitation sent the populations in characteristic directions. Mutation supply and the fitness contribution of mutations were estimated and demonstrated to be potential influences in the choice of particular adaptation pathways under severe and mild glucose limitation. Predicting all the mutations fixed in adapting populations is beyond our current understanding of evolutionary processes, but the interplay between ancestor physiology and the initiation of adaptation pathways is demonstrated and definable in bacterial populations. PMID- 12737664 TI - Estimating average cellular turnover from 5-bromo-2'-deoxyuridine (BrdU) measurements. AB - Cellular turnover rates in the immune system can be determined by labelling dividing cells with 5-bromo-2'-deoxyuridine (BrdU) or deuterated glucose ((2)H glucose). To estimate the turnover rate from such measurements one has to fit a particular mathematical model to the data. The biological assumptions underlying various models developed for this purpose are controversial. Here, we fit a series of different models to BrdU data on CD4(+) T cells from SIV(-) and SIV(+) rhesus macaques. We first show that the parameter estimates obtained using these models depend strongly on the details of the model. To resolve this lack of generality we introduce a new parameter for each model, the 'average turnover rate', defined as the cellular death rate averaged over all subpopulations in the model. We show that very different models yield similar estimates of the average turnover rate, i.e. ca. 1% day(-1) in uninfected monkeys and ca. 2% day(-1) in SIV-infected monkeys. Thus, we show that one can use BrdU data from a possibly heterogeneous population of cells to estimate the average turnover rate of that population in a robust manner. PMID- 12737665 TI - The role of ultraviolet-A reflectance and ultraviolet-A induced fluorescence in the appearance of budgerigar plumage: insights from spectrofluorometry and reflectance spectrophotometry. AB - Fluorescence has so far been found in 52 parrot species when illuminated with ultraviolet-A (UVA) 'black' lamps, and two attempts have been made to determine whether such fluorescence plays any role in sexual signalling. However, the contribution of the reflectance versus fluorescence to the total radiance from feathers, even in the most studied species to date (budgerigars), is unclear. Nor has the plumage of this study species been systematically assessed to determine the distribution of fluorescent patches. We therefore used spectrofluorometry to determine which areas of budgerigars fluoresce and the excitation and emission spectra involved; this is the first time that such a technique has been applied to avian plumage. We found that both the yellow crown and (normally hidden) white downy chest feathers exhibit strong UVA-induced fluorescence, with peak emissions at 527 nm and 436 nm, respectively. Conversely, the bright-green chest and dark blue tail feathers do not fluoresce. When comparing reflectance spectra (400-700 nm) from the yellow crown using illuminants with a proportion of UVA comparable to daylight, and illuminants with all UVA removed, no measurable difference resulting from fluorescence was found. This suggests that under normal daylight the contribution of fluorescence to radiance is probably trivial. Furthermore, these spectra revealed that males had fluorescent crowns with substantially higher reflectance than those of females, in both the UV waveband and at longer wavelengths. Reflectance spectrophotometry was also performed on a number of live wild-type male budgerigars to investigate the chromatic contrast between the different plumage areas. This showed that many plumage regions are highly UV reflective. Overall our results suggest that rapid surveys using UVA black lamps may overestimate the contribution of fluorescence to plumage coloration, and that any signalling role of fluorescence emissions, at least from the yellow crown of budgerigars, may not be as important as previously thought. PMID- 12737666 TI - Diversification and cumulative evolution in New Caledonian crow tool manufacture. AB - Many animals use tools but only humans are generally considered to have the cognitive sophistication required for cumulative technological evolution. Three important characteristics of cumulative technological evolution are: (i) the diversification of tool design; (ii) cumulative change; and (iii) high-fidelity social transmission. We present evidence that crows have diversified and cumulatively changed the design of their pandanus tools. In 2000 we carried out an intensive survey in New Caledonia to establish the geographical variation in the manufacture of these tools. We documented the shapes of 5550 tools from 21 sites throughout the range of pandanus tool manufacture. We found three distinct pandanus tool designs: wide tools, narrow tools and stepped tools. The lack of ecological correlates of the three tool designs and their different, continuous and overlapping geographical distributions make it unlikely that they evolved independently. The similarities in the manufacture method of each design further suggest that pandanus tools have gone through a process of cumulative change from a common historical origin. We propose a plausible scenario for this rudimentary cumulative evolution. PMID- 12737667 TI - Plankton blooms induced by turbulent flows. AB - Plankton play an important role in the ecology of the ocean and the climate because of their participation in the global carbon cycle at the base of the food chain. However, damaging plankton blooms can sometimes occur and are initially characterized by sudden transient increases in the phytoplankton population. They are thought to be driven by several effects, such as seasonal variations in temperature and salinity, and nutrient mixing. Furthermore, phytoplankton and zooplankton have different buoyancy properties, leading to a differential response in turbulent environments. In this paper, we investigate this effect in a model of advected plankton dynamics. We find that, over a range of parameter values, flows of marine species subjected to inertial/viscous forces naturally lead to patchiness and, in turn, periodically sustained plankton blooms. PMID- 12737668 TI - Effect of habitat fragmentation on levels and patterns of genetic diversity in natural populations of the peat moss Polytrichum commune. AB - Peat bogs represent unique ecosystems that are under particular threat from fragmentation due to peat harvesting, with only 38% of the original peatland in Europe remaining intact and unaffected by peat cutting, drainage and silviculture. In this study, we have used microsatellite markers to determine levels and patterns of genetic diversity in both cut and uncut natural populations of the peat moss Polytrichum commune. Overall diversity levels suggest that there is more genetic variation present than had previously been assumed for bryophytes. Despite this, diversity values from completely cut bogs were found to be lower than those from uncut peatlands (average 0.729 versus 0.880). In addition, the genetic diversity was more highly structured in the cut populations, further suggesting that genetic drift is already affecting genetic diversity in peat bogs subjected to fragmentation. PMID- 12737669 TI - Ovarian hyperstimulation syndrome: distinction between local and systemic disease. AB - The ovarian hyperstimulation syndrome (OHSS) is an iatrogenic, unpredictable and potentially life-threatening complication in patients submitted to pharmacological ovarian stimulation. Information on risk factors, etiopathogenetic mechanisms, prevention strategies and therapeutic management is continuously updated. The present study retrospectively analyzed 123 women affected by different grades of OHSS as a result of pharmacological ovulation induction. Hospital admission was suggested in 14 patients with severe OHSS, whereas patients with moderate or mild OHSS were followed in the out-patient section of our department. The results confirmed the efficacy of the therapeutic scheme adopted. The syndrome is localized to the ovaries at the time that the condition is triggered; when organs different from the ovaries become involved, OHSS assumes systemic aspects. The different clinical signs are the basis of a proposal of a local and systemic classification. PMID- 12737670 TI - Does pre-treatment with micronized progesterone affect the ovarian response to a gonadotropin releasing hormone agonist flare-up protocol? AB - The purpose of this study was to investigate the ovarian response and the receptivity of the endometrium in women pre-treated with micronized progesterone. Eighty-two normogonodotropic women undergoing in vitro fertilization were studied. Thirty received micronized progesterone 1500 mg/day from day 21 of the cycle for a minimum of 2 weeks, and 52 did not receive micronized progesterone (control group). A gonadotropin releasing hormone agonist (GnRH-a) was administered to all the patients in the follicular phase (flare-up). Twenty-five cycles were cancelled for fertilization failure due to male factor, 12 (40%) in the progesterone group and 13 (25%) in the control group (p = 0.271). There was no difference in the number of oocytes retrieved (7.3 +/- 5 vs. 8.2 +/- 4), fertilization rate (50.8% vs. 65%), clinical pregnancy rate (16.6% vs. 25%) or implantation rate (8% vs. 14%). In the progesterone group cases without fertilization, we performed two biopsies to evaluate the receptivity of the endometrium. Pinopode expression was noted 7 days after oocyte retrieval. It seems that the administration of micronized progesterone in the previous cycle does not affect the ovarian response to the combination of follicular phase GnRH a and gonadotropins, nor the receptivity of the endometrium. PMID- 12737671 TI - A meta-analysis on the correlation between ovarian activity and the incidence of intermenstrual bleeding during low-dose oral contraceptive use. AB - We aimed to evaluate potential correlation between ovarian activity during use of combined oral contraceptives and the incidence of intermenstrual bleeding. Data from seven prospective clinical studies with five different combined low-dose oral contraceptives were retrospectively analyzed to determine ovarian activity measured by the Hoogland Score (follicle diameter and endogenous hormone levels) and cycle control. A total of 227 young fertile women were evaluated over three treatment cycles each. Women with intermenstrual bleeding had statistically significantly higher estradiol levels than those without intermenstrual bleeding. Also, women with intermenstrual bleeding had significantly larger follicle-like structures than those without intermenstrual bleeding. For example, in the second treatment cycle the difference of the mean follicle diameters between women without intermenstrual bleeding (12.5 mm) and women with spotting (16.9 mm) or breakthrough bleeding (16.1 mm) was statistically significant (p = 0.0179). Less than 17% of women with Hoogland Score 1, 2 or 3 (low ovarian activity) reported intermenstrual bleeding. On the other hand, 35.2% of women with Hoogland Score 4 (active follicle-like structures) reported intermenstrual bleeding. The association between bleeding and Hoogland Score was statistically significant (p < 0.0011). The findings of this retrospective analysis provide evidence that high ovarian suppression is positively correlated with improved cycle control in terms of less frequent intermenstrual bleeding - slight and heavy. PMID- 12737673 TI - Inhibin B and ovarian function after total abdominal hysterectomy in women of reproductive age. AB - The aim of this prospective study was to assess ovarian function using clinical and endocrine parameters in women of reproductive age who underwent total abdominal hysterectomy. Sixty-one women, aged 40 mIU/ml, estradiol of < 20 pg/ml and inhibin B of < 5 ng/ml, compatible with ovarian failure. In the control group, all the parameters studied remained unchanged. These results suggest that total abdominal hysterectomy accelerates the decline in ovarian function in women of reproductive age. PMID- 12737672 TI - Pinealectomy changes rat ovarian interstitial cell morphology and decreases progesterone receptor expression. AB - The aim of this study was to evaluate the rat ovarian morphological and function changes after pinealectomy (px). Two months after px, young female Wistar rats were sacrificed and the right ovaries were analysed morphologically and the left ovaries were used for steroid receptor binding experiments. Blood was collected and steroid hormone and melatonin levels were measured using radioimmunoassay kits. Results revealed that in the px group the rat ovaries had an increase in the number of atretic follicles and interstitial cells. These cells showed hyperactivity features on transmission electron microscopy and morphometric analysis (p < 0.05 compared with control and sham groups). Px-group serum showed an increase in estradiol (p < 0.05) and a decrease in progesterone levels (p < 0.05) compared with other groups. Moreover, progesterone receptor expression was lower than control and sham groups (p < 0.05). We postulate that pinealectomy leads to many morphological alterations of rat ovaries that are associated with functional changes in steroidogenesis and a decrease in progesterone receptor expression. PMID- 12737674 TI - Maternal low birth weight and gestational hyperglycemia. AB - We aimed to investigate whether birth weight could predict the subsequent risk of gestational diabetes and impaired glucose tolerance. Consecutive women with a singleton pregnancy and gestational diabetes (n = 50), impaired glucose tolerance (n = 50) and normoglycemia (n = 200) were included in the study. Birth data were collected from original hospital records of the women. Women with gestational hyperglycemia were significantly older and heavier than those with normoglycemia. Maternal birth weights significantly declined for each class of glucose tolerance (3389 +/- 644; 3184 +/- 583 and 3077 +/- 661, respectively for women with normoglycemia, impaired glucose tolerance and gestational diabetes). After adjustment for age, gestational age and weight gain, maternal diabetes, and pre pregnancy body mass index, maternal birth weight was negatively related to impaired glucose tolerance (OR 0.88, 95% CI 0.81-0.97) and to gestational diabetes (OR 0.82, 95% CI 0.74-0.91) in a multiple logistic regression model. These findings suggest that women with low birth weight constitute a group at increased risk for both gestational impaired glucose tolerance and diabetes. PMID- 12737675 TI - Insulin secretion and insulin sensitivity in normal pregnancy and gestational diabetes mellitus. AB - The purpose of this study was to evaluate insulin sensitivity, beta-cell function and islet-cell-directed autoimmunity in pregnant women with normal glucose tolerance and gestational diabetes mellitus (GDM). A total of 21 women with normal glucose tolerance and 21 women with GDM were evaluated at 24-36 weeks' gestation. Insulin resistance and beta-cell function were evaluated using the continuous infusion of glucose with model assessment (CIGMA) method, which aims to give a near-physiological stimulus and to evaluate the endogenous insulin and glucose response. Islet-cell autoantibody was positive in one woman with GDM, and glutamic acid decarboxylase autoantibodies were negative in both groups. The calculated CIGMA insulin resistance (CIGMA IR) was 2.04 +/- 1.74 and 1.08 +/- 1.22 in patients with GDM and in control subjects, respectively (p < 0.05). CIGMA percentage beta-cell values were 64.04 +/- 44.55% and 87.07 +/- 52.77% in patients with GDM and control subjects, respectively (p > 0.05). Decreased insulin sensitivity in late pregnancy was more evident in lean GDM subjects with mild hyperglycemia who did not require insulin therapy, and beta-cell function was partially preserved in this group of patients. PMID- 12737676 TI - Increased vascular surface density in ovarian endometriosis. AB - Our goal in this study was to investigate the presence of angiogenesis-related factors in endometriomas by evaluating their vascular surface densities. Thirty ovarian samples were included in the study. Of these, ten were histologically confirmed endometriomas, ten were ovarian specimens in the follicular phase and ten were ovarian specimens in the luteal phase, serving as controls. Histological specimens were immunostained for von Willebrand factor (vWF: factor VIII-related antigen) and CD34. The area with the highest microvessel density in endometriosis and in the normal ovary was evaluated by using an intercept grid. All microvessels in a specific field (x 100 magnification) were counted and vascular surface density was measured, as 164.01 +/- 21.26 vs. 125.15 +/- 11.28 and 117.44 +/- 9.27 by using vWF, and as 172.97 +/- 25.64 vs. 138.65 +/- 32.21 and 120.34 +/ 18.40 by using CD34 in endometriotic, follicular and luteal ovarian samples, respectively (p < 0.001). The mean vascular surface density was significantly higher in endometriosis than in the ovarian samples of the follicular phase or the luteal phase. No significant difference was seen between normal ovarian samples. Endometriosis was associated with angiogenic properties. Having demonstrated elevated angiogenic factors in endometriotic samples, we concluded that activation of angiogenesis might be a key factor in the pathogenesis of endometriosis. PMID- 12737677 TI - Adrenal function under long-term raloxifene administration. AB - The aim of the present study was to evaluate the effect of long-term (12 months) administration of raloxifene hydrochloride (60 mg/day) on the steroid production of the adrenal cortex and on the hypothalamic-pituitary-adrenal axis in postmenopausal women. We performed a basal evaluation, a corticotropin releasing factor (CRF) (100 microg i.v. bolus) test and a dexamethasone (DXM) (0.25 mg) suppression-adrenocorticotropic hormone (ACTH) (10 microg i.v. bolus) stimulation test in 11 postmenopausal women, before and after 3, 6 and 12 months of raloxifene treatment. Raloxifene administration significantly modified circulating levels of adrenal steroids, decreasing cortisol (-24%), dehydroepiandrosterone (DHEA) (-36%), and its sulfate (DHEAS) (-41%), and androstenedione (-29%), and increasing circulating allopregnanolone (+39%) levels. Progesterone and 17OH-progesterone levels remained unmodified, while estradiol and estrone levels showed a significant decrease (-51% for estradiol and -61% for estrone). We also observed an increase in circulating ACTH (+58%) and beta-endorphin (+120%). No modifications in the hormonal responses to CRF were observed during the treatment. DXM significantly suppressed circulating steroids at any time with a lower suppression of cortisol from the third month and a higher suppression of DHEA at 12 months. ACTH administration was associated with a significantly blunted cortisol response from the sixth month and a significantly increased response of allopregnanolone from the third month. The present data exclude a raloxifene effect on pituitary sensitivity to CRF and demonstrate a reduced adrenal sensitivity to ACTH, sustained by the opposite changes in basal cortisol and Delta5 androgens, which were reduced, and in ACTH and beta-endorphin, which were increased, as well by the reduced response of cortisol to the direct ACTH stimulus. The reduction of circulating cortisol levels and cortisol response to the ACTH challenge suggests that raloxifene protects against the neurotoxic effects of endogenous glucocorticoids. Furthermore, the progressive increase in basal allopregnanolone and its increased response to ACTH indicate that chronic raloxifene administration exerts direct effects on the pattern of adrenal enzymes, leading to specific changes in the circulating levels of this anxiolytic progesterone metabolite. The important reduction in the circulating levels of estradiol and estrone under long-term raloxifene administration may represent a further mechanism by which this molecule may exert a protective effect against breast and endometrial malignancies. PMID- 12737678 TI - Estrogen receptor knockout mice: phenotypes in the female reproductive tract. AB - There is a broad spectrum of organ systems that respond to estrogen hormones, including the female and male reproductive tracts, mammary gland, the skeleton, cardiovascular system and central nervous system. The physiological effects of estrogens are mediated by the estrogen receptor, a member of the nuclear receptor superfamily of transcription factors. Two estrogen receptors have been identified: the originally described estrogen receptor alpha (ERalpha) and the more recently discovered estrogen receptor beta (ERbeta). Three different estrogen receptor knockout (ERKO) mouse models were generated, carrying a null mutation in the ERalpha gene (alphaERKO), the ERbeta gene (betaERKO) or both genes (alphabetaERKO). The generation of the different ERKO mice provides ideal models for studying the physiological consequences of the complete lack of estrogen receptor activity and the distinct roles of both estrogen receptors in various tissues. This review summarizes the phenotypes principally seen in the female reproductive system of the different ERKO mice. PMID- 12737679 TI - Inhaled corticosteroids in chronic obstructive pulmonary disease: new trials and old practices. AB - The practice of using inhaled steroids (ICS) in chronic obstructive pulmonary disease (COPD) is common but controversial. Four large randomized controlled trials provide an objective assessment of this practice. An examination of the design of these studies is useful to explaining some of the differences between their results but may also help to appropriately integrate the findings of these studies into clinical practice. All studies agree that the early introduction of the inhaled corticosteroids (ICS) does not appear to affect the rate of decline in FEV1. Thus, the routine prescription of these agents to asymptomatic patients with well-preserved lung function is not indicated. However, more selective use of ICS in patients with moderately severe disease (FEV1 < 50% predicted) may produce clinical benefit as measured by an increase in FEV1, reduced symptoms and fewer exacerbations. The effectiveness of ICS in patients with severe disease (FEV < 800 mL, or chronic respiratory failure) has not been studied due to practical difficulties of enrolling and maintaining such patients in clinical trials. Similarly patients with evidence of chronic bronchitis and emphysema who demonstrate high degrees of reversibility (>10% predicted FEV1) are usually excluded from both COPD and asthma trials, and the effectiveness of ICS in this population is also not known. Neither responsiveness to a single dose of beta(2) agonist nor to a 14-day trial of systemic steroids has been shown to be a reliable predictor of response to ICS. For individual patients with moderate or severe symptomatic COPD, an 8-week therapeutic trial with ICS may be needed to assess clinical benefit. PMID- 12737680 TI - A comparison of planar scintigraphy and SPECT measurement of total lung deposition of inhaled aerosol. AB - Planar gamma camera imaging of inhaled aerosol deposition is extensively used to assess the total deposition in the lung. However, validation of the measurements is not straightforward, as gold standard measurements of lung activity against which to compare are not readily available. Quantitative SPECT imaging provides an alternative method for comparison. Four different methods for planar image quantification are compared. Two attenuation correction techniques, thickness measurement and transmission measurement, have been combined with two scatter correction techniques, reduced attenuation coefficient and line-source scatter function convolution subtraction. Each technique has been applied to 10 studies of aerosol deposition of a fine aerosol (mass median aerodynamic diameter 1.8 microm) and 10 studies using a coarse aerosol (mass median aerodynamic diameter 6.5 microm). The total activity in the right lung for each measurement has been compared to the value determined from SPECT imaging on the same subjects. When the thickness measurement and transmission techniques were applied with scatter compensation using a reduced attenuation coefficient, activity was systematically overestimated by 5% in both cases. The corresponding random errors (coefficient of variation) were 8.6% and 6.6%. Separate scatter correction reduced these systemic errors significantly to -1.5% and 2.7%, respectively. The random errors were not affected. All techniques provided assessment of total lung activity with an accuracy and precision that differed by less than 10% compared to the SPECT values. Planar gamma camera imaging provides a good method of assessing total lung deposition of inhaled aerosol. PMID- 12737681 TI - The inhalation manager: a new computer-based device to assess inhalation technique and drug delivery to the patient. AB - The rational choice of an inhalation device is a cornerstone in the effective management of asthma and COPD. In this publication, we describe the development of a new system, the Inhalation Manager, which, for the first time, offers the possibility to assess the entire inhalation maneuver of patients using original devices under everyday conditions. So far the Inhalation Manager allows the measurement of inspiratory maneuvers of patients through placebo inhalation devices of the most common breath-actuated CFC-free inhalers in the market for the three main glucocorticosteroids Budesonide [Turbohaler (TH), dry powder inhaler (DPI)], Beclomethasone dipropionate [Autohaler (AH), breath-actuated pressurized metered dose inhaler (pMDI)], and Fluticasone propionate [Diskus (DI), DPI] by means of a pneumotachometer. In addition, it allows allocation of the individual maneuver to the expected drug delivery values (mass output and particle size distribution) of these three devices. In a field trial, the inhalation technique of 628 (TH), 794 (AH), and 795 (DI) patients, respectively, was tested in 72 pulmonologist practices with the Inhalation Manager. For patients in the 18-59-year-old group, the Inhalation Manager detected the following percentages needing improvement: 1.5% for the Autohaler device, 16.7% for the Diskus, and 38.9% for the Turbohaler. In the 60-99-year-old group, percentages needing improvement were 1.5%, 31.5%, and 66.1% for the Autohaler, Diskus, and Turbohaler, respectively. Therefore, the Inhalation Manager could become an essential tool in asthma management by finding the most suitable inhaler for an individual patient and by training the optimal inhalation technique. PMID- 12737682 TI - Assessment of adrenal suppression from two new dry powder inhaler formulations of budesonide delivered by Clickhaler compared with the Pulmicort Turbuhaler. AB - Assessment of adrenal suppression via systemic cortisol levels provides an indirect measure of the lung delivery of inhaled corticosteroids. This randomized, placebo-controlled, double-blind, double-dummy crossover study compared urinary and plasma cortisol levels in healthy adult volunteers following single 1,000-microg doses of budesonide from two multiple dose dry powder inhalers (DPIs). Two new formulations of budesonide (lactose and PassCal) delivered from the Clickhaler were compared with Pulmicort from the Turbuhaler. An open dose (2,000 microg) of Pulmicort Turbuhaler was included to validate the experimental model. Overnight (22:00-07:00 h) and early morning (07:00-08:00 h) urine and 08:00 h plasma samples were collected after each treatment and cortisol levels analyzed by radioimmunoassay. Combined overnight and early morning urinary cortisol values for PassCal Clickhaler and Pulmicort Turbuhaler (1,000 microg) were statistically significantly lower than placebo (p < 0.05). The lactose budesonide Clickhaler showed a non-significant urinary cortisol reduction compared with placebo. Differences between the three 1,000-microg budesonide treatments were not significant. The Pulmicort Turbuhaler 2,000 microg showed significant urinary cortisol suppression compared with placebo. Plasma cortisol showed similar effects, with significance between the two Pulmicort doses. These results suggest that adrenal suppression can be used to assess the pulmonary bioavailability of different formulation. PMID- 12737683 TI - Lung deposition in cystic fibrosis patients using an ultrasonic or a jet nebulizer. AB - This study was conducted to compare pulmonary deposition following inhalation with an ultrasonic and a jet nebulizer in CF patients under conditions relevant to practice. The marker substance used to estimate the relative lung bioavailability was sodium cromoglycate (SCG), which is poorly absorbed from the gastrointestinal tract, but is completely absorbed from the lungs. Ten CF patients (aged 9-21 years) used an ultrasonic nebulizer (Multisonic compact 2.4 MHz) and a jet nebulizer (Parimaster, LC Plus Turbo) in a crossover design to inhale a solution containing 20 mg of SCG and a beta(2)-agonist. Urine was collected in five fractions until 12 h p. a., and the excreted SCG was determined by means of HPLC. Prior to each inhalation, the patients' pulmonary function was measured employing a Pneumoscope. Using the ultrasonic nebulizer, the amount of SCG excreted in urine was significantly greater than that after inhalation with the jet nebulizer (1.43 +/- 0.47 mg vs. 1.04 +/- 0.47 mg; p = 0.002), despite the larger residual volume in the ultrasonic nebulizer. The absorption half-life for SCG following ultrasonic nebulization was significantly shorter when compared with jet nebulization (84 +/- 14 min vs. 101 +/- 19 min; p = 0.005), being suggestive of a more peripheral deposition. Furthermore, an inverse relationship was found between absorption half-life and FEV(1) (% pred.) (r = -0.655, p = 0.04) or MMEF(75/25) (% pred.) (r = -0.844, p = 0.031), but only with the ultrasonic nebulizer. In conclusion, the ultrasonic nebulizer tested when used for inhalation in CF patients was found to be at least equivalent to the jet nebulizer. PMID- 12737684 TI - Effect of propellant on the pharmacokinetics and pharmacodynamics of inhaled albuterol in asthmatic subjects. AB - Hydrofluoroalkane (HFA) propellants have largely replaced chlorofluorocarbon (CFC) propellants in metered dose inhalers (MDI). It is important to document the pharmacokinetics (PK) and pharmacodynamics (PD) of medications delivered using HFA propellants compared to CFC propellants. Six adult asthmatics with mild to moderate asthma were selected for the study. Each subject inhaled 180 microg of albuterol from an MDI with holding chamber. Venous blood was collected for measuring albuterol levels at intervals over 12 h, and spirometric measurements of airflow were measured over the same time period. Plasma samples were analyzed using a GC/MS assay developed in our laboratory. PK and PD parameters were calculated by nonlinear regression using WinNonlin. There were no statistically significant differences between PD parameters for HFA versus CFC propellants. The area under the plasma albuterol concentration versus time curve (AUC) was 72% greater for the HFA formulation, indicating a greater lung bioavailability (p = 0.015). This difference in bioavailability did not result in a statistically significant difference in FEV(1) values between the two propellants. PMID- 12737685 TI - Evaluation of the survival of bacterial contaminants in an inhalable insulin powder. AB - Survival and growth of three model test bacterial species (Pseudomonas fluorescens, Staphylococcus epidermidis and Bacillus subtilis), present in the air and/or in the human respiratory tract, were tested in inhalable insulin lactose powder under optimal relative humidity and temperature conditions (RH = 96% and optimal growth temperature for each bacterium of 26-37 degrees C) as well as representative indoor conditions (RH = 43% and T = 20 degrees C). The bacteria survived from 12 h to 7 days depending on the bacterial species and the test condition. P. fluorescens vegetative cells had the lowest and B. subtilis spores the highest survival rate. It was found that insulin-lactose powder does not support bacterial growth and that higher bacterial survival rate was found under representative indoor conditions. Selected experiments were performed with B. subtilis by adding sterile saliva into insulin-lactose powder to represent a typical condition for inhaler use. Furthermore, two other powders were tested with B. subtilis: one representing an inert powder without any nutrients (glass beads) and the other representing a powder with optimal nutrients (tryptic soy broth powder). The data indicate that the survival rate of B. subtilis did not change after the saliva was added and that the survival in insulin-lactose powder was similar to that in inert powder but lower than in powder with optimal nutrients. These results suggest that bacterial growth on residual powder in the inhaler under patient use conditions is unlikely and therefore the concern for patient safety is remote. PMID- 12737686 TI - Particle characteristics and lung deposition patterns in a human airway replica of a dry powder formulation of polylactic acid produced using supercritical fluid technology. AB - Polylactic acid (PLA) powders have been used as vector particles to carry pharmaceutical material. Drugs incorporated in the PLA powder can be retained in the lung for a longer period and may be more effective than free-form drugs. A new formulation of L-PLA dry powder, which was easy to disperse in the air, was produced by using a supercritical technology. The L-PLA powder was characterized in terms of physical particle size and aerodynamic size as generated with a Turbuhaler dry powder inhaler (DPI). Electron microscopy analysis of the particles indicated that they were individual particles in bulk form and became aggregate particles after generation by the Turbuhaler. Aerodynamic particle size analysis using both an Aerodynamic Particle Sizer (APS) aerosol spectrometer and Andersen impactor showed that the aerodynamic size decreased as the flow rate in the Turbuhaler increased from 28.3 to 90 L min(-1). Deposition patterns in the human respiratory tract were estimated using a realistic physical replica of human airways. Deposition of the L-PLA was high (80.8%) in the oral airway at 28.3 L min(-1) and an average of 73.4% at flow rates of 60 and 90 L min(-1). In the lung region, the deposition totaled 7.2% at 28.3 L min(-1), 18.3% at 60 L min(-1), and 17.6% at 90 L min(-1). These deposition patterns were consistent with aerodynamic size measurement, which showed 76 to 86% deposition in the USP/EP (US Pharmacopoeia/European Pharmacopoeia) induction port. As the flow rate increased, fewer aggregates were formed resulting in the smaller aerodynamic particles. As a result, more particles penetrated the oral airways and were available for deposition in the lung. Our results showed that L-PLA particles as manufactured by the supercritical technology could be used in a DPI that does not require the use of carrier particles to facilitate aerosol delivery. PMID- 12737690 TI - For fear of pain: British surgery, 1790-1850. PMID- 12737707 TI - Toward optimal health: the experts discuss the diet debate. Interview by Jodi Godfrey Meisler. PMID- 12737708 TI - Nutrition and Women's Health Symposium. Introduction. PMID- 12737709 TI - Dietary fat and cardiovascular disease risk: quantity or quality? AB - When considering dietary fat quantity, there are two main factors to consider, impact on body weight and plasma lipoprotein profiles. Data supporting a major role of dietary fat quantity in determining body weight are weak and may be confounded by differences in energy density, dietary fiber, and dietary protein. With respect to plasma lipoprotein profiles, relatively consistent evidence indicates that under isoweight conditions, decreasing the total fat content of the diet causes an increase in triglyceride and decrease in high-density lipoprotein (HDL) cholesterol levels. When considering dietary fat quality, current evidence suggests that saturated fatty acids tend to increase low-density lipoprotein (LDL) cholesterol levels, whereas monounsaturated and polyunsaturated fatty acids tend to decrease LDL cholesterol levels. Long-chain omega-3 fatty acids, eicosapentaenoic acid (EPA) (20:5n-3) and docosahexaenoic acid (DHA) (22:6n-3), are associated with decreased triglyceride levels in hypertriglyceridemic patients and decreased risk of developing coronary heart disease (CHD). Dietary trans-fatty acids are associated with increased LDL cholesterol levels. Hence, a diet low in saturated and trans-fatty acids, with adequate amounts of monounsaturated and polyunsaturated fatty acids, especially long-chain omega-3 fatty acids, would be recommended to reduce the risk of developing CHD. Additionally, the current data suggest it is necessary to go beyond dietary fat, regardless of whether the emphasis is on quantity or quality, and consider lifestyle. This would include encouraging abstinence from smoking, habitual physical activity, avoidance of weight gain with age, and responsible limited alcohol intake (one drink for females and two drinks for males per day). PMID- 12737710 TI - Types of carbohydrates and risk of cardiovascular disease. AB - The purpose of this review is to provide an overview of the role of dietary carbohydrates in the etiology of cardiovascular disease (CVD) among women. Many factors are thought to affect insulin resistance, and little is known about the role of diet. Through effects on postprandial glucose and insulin, dietary glycemic load may have an important role in the insulin resistance syndrome (IRS). Dietary fiber, through its influence on the glycemic load or through other pathways, may also have important effects on this syndrome. Many short-term experimental studies have supported these hypotheses. Interestingly, associations may be stronger among overweight individuals than among nonoverweight individuals. Similar to findings for fruits and vegetables, whole grain intake has been found to be consistently associated with a reduction in risk of coronary heart disease (CHD) among both men and women. Several large randomized trials of primary and secondary prevention to date have demonstrated the efficacy of diets based on an abundance of plant foods and, therefore, high carbohydrate quality. The recommendations to follow a diet including an abundance of fiber-rich foods in order to prevent CVD and diabetes are based on a wealth of consistent scientific evidence. More long-term controlled trials are needed to improve our understanding of efficacy and mechanisms. Women and a variety of racial/ethnic groups should be represented in these studies whenever possible. PMID- 12737711 TI - Normal bone accretion and effects of nutritional disorders in childhood. AB - Although osteoporosis is usually considered to be a health concern of the elderly, increased attention is being paid to children and adolescents who are at risk for developing this devastating disease. As osteoporosis is a preventable condition with no identified cure, focus has been placed on modifiable areas in a young person's life that may prevent the development of the disease. A child or adolescent's nutrition is an example of such an area. This review examines factors influencing normal bone development and emphasizes the importance of the adolescent years as a time for peak bone accretion. Current methods to evaluate skeletal status are examined, including the challenges that arise in interpreting bone densities in children who have growing bones. Children and adolescents who are at high risk for osteoporosis are discussed, with an emphasis on groups in whom poor nutrition likely mediates bone loss. Two models of malnutrition, anorexia nervosa and the female athlete triad, are discussed, with emphasis on the way in which each has deleterious effects on the adolescent skeleton. The promotion of skeletal health is ultimately the encouragement of good general health principles for all young people. Approaches for working with children and adolescents are reviewed, including literature supporting each strategy. PMID- 12737712 TI - Mechanisms by which nutritional disorders cause reduced bone mass in adults. AB - Nutritional disorders that cause bone loss in adults include disordered eating behaviors (female athlete triad and anorexia nervosa), gastrointestinal diseases (celiac sprue, inflammatory bowel disease, and other malabsorption syndromes), alcoholism, and hypervitaminosis A. These disorders exert their effects on bone through a number of mechanisms, including estrogen deficiency. Deficiencies of anabolic hormones may also be important, including insulin-like growth factor I (IGF-I), a nutritionally regulated bone trophic factor. In addition, low weight itself is a risk factor for bone loss and decreased bone formation. Reduced calcium and vitamin D availability, with resultant secondary hyperparathyroidism, is another important mechanism of bone loss in nutritional disorders. This review discusses nutritional causes of reduced bone mass in adults and how nutritional disorders exert deleterious effects on the skeleton. PMID- 12737713 TI - Vitamin D and bone health in postmenopausal women. AB - Osteoporosis, a disease of increased skeletal fragility, is becoming increasingly common as the U.S. population ages. Adequate vitamin D and calcium intake is the cornerstone of osteoporosis prevention and treatment. Age-related changes in vitamin D and calcium metabolism increase the risk of vitamin D insufficiency and secondary hyperparathyroidism. Although longitudinal data have suggested a role of vitamin D intake in modulating bone loss in perimenopausal women, studies of vitamin D and calcium supplementation have failed to support a significant effect of vitamin D and calcium during early menopause. There is a clearer benefit in vitamin D and calcium supplementation in older postmenopausal women. Vitamin D intake between 500 and 800 IU daily, with or without calcium supplementation, has been shown to increase bone mineral density (BMD) in women with a mean age of approximately 63 years. In women older than 65, there is even more benefit with vitamin D intakes of between 800 and 900 IU daily and 1200-1300 mg of calcium daily, with increased bone density, decreased bone turnover, and decreased nonvertebral fractures. The decreases in nonvertebral fractures may also be influenced by vitamin D-mediated decreases in body sway and fall risk. There are insufficient available data supporting a benefit from vitamin D supplementation alone, without calcium, to prevent osteoporotic fracture in postmenopausal women. PMID- 12737714 TI - Early life predictors of chronic disease. AB - Many chronic diseases may have their seeds early in life. Observations from the Dutch famine have taught us that the intrauterine environment is an important determinant of adult health. Birth weight has been related to cardiovascular disease (CVD), hypertension, diabetes, and cancer. Critical phases for adult obesity development include time periods between conception and adolescence. A life course approach to chronic disease prevention includes the study of maternal diet during pregnancy, intrauterine exposures, postnatal growth, and the adolescent period. PMID- 12737715 TI - Overweight and obesity in women: health risks and consequences. AB - The evidence for the adverse effects of obesity on women's health is overwhelming and indisputable. Obesity, especially abdominal obesity, is central to the metabolic syndrome and is strongly related to polycystic ovary syndrome (PCOS) in women. Obese women are particularly susceptible to diabetes, and diabetes, in turn, puts women at dramatically increased risk of cardiovascular disease (CVD). Obesity substantially increases the risk of several major cancers in women, especially postmenopausal breast cancer and endometrial cancer. Overweight and obesity are associated with elevated mortality from all causes in both men and women, and the risk of death rises with increasing weight. Curbing the twin epidemics of obesity and diabetes calls for not only changes in diet and lifestyle at individual levels but also changes in policy, physical and social environment, and cultural norms. PMID- 12737716 TI - Diet, body weight, and colorectal cancer: a summary of the epidemiologic evidence. AB - Colorectal cancer is the second leading cause of cancer death in the United States, and the number of new cases annually is approximately equal for men and women. Several nutritional factors are likely to have a major influence on risk of this cancer. Physical inactivity and excessive adiposity, especially if centrally distributed, clearly increase the risk of colon cancer. Hyperinsulinemia may be an important underlying risk factor. In conjunction with obesity and physical inactivity, which induce a state of insulin resistance, certain dietary patterns that stimulate insulin secretion, including high intakes of red and processed meats, saturated and trans-fats, and highly processed carbohydrates and sugars, may increase the risk of colon cancer. There is evidence suggesting that some component of red meat may independently increase the risk of colorectal cancer, and some micronutrients may be important as protective agents. Currently, the evidence is strongest for folate and calcium. Folate may be especially important in alcohol drinkers because alcohol appears to increase the risk, particularly when folate intake is low. This interaction may be related to the antifolate properties of alcohol. In contrast to earlier studies, more recent epidemiologic studies have generally not supported a strong influence of dietary fiber or fruits and vegetables, although these have other health benefits, and their consumption should be encouraged. The majority of colon cancers, as well as many other conditions, may be prevented by lifestyle alterations in the intake of these nutritional factors, in addition to other factors, such as smoking. PMID- 12737717 TI - Diet and breast cancer. AB - Obesity, overweight, and a sedentary lifestyle-all common conditions in breast cancer patients-are likely to be associated with poor survival and poor quality of life in women with breast cancer. Diet-related factors are thought to account for about 30% of cancers in developed countries. Most studies of diet and healthcare have focused on the role of single nutrients, foods, or food groups in disease prevention or promotion. Recent cancer guidelines on nutrition and physical activity emphasize diets that promote maintenance of a healthy body weight and a prudent dietary pattern that is low in red and processed meats and high in a variety of vegetables, fruits, and whole grains. Except for dietary fat, few nutritional factors in adult life have been associated with breast cancer. Extensive data from animal model research, international correlations linking fat intake and breast cancer rates, and case-control studies support the hypothesis that a high-fat diet is conducive to the development of breast cancer in postmenopausal women. Conflicting findings from cohort studies, however, have created uncertainty over the role of dietary fat in breast cancer growth and recurrence. Results from large-scale nutritional intervention trials are expected to resolve such issues. As new and improved data on dietary factors and patterns accumulate, dietary guidelines for cancer risk reduction will become more focused. PMID- 12737719 TI - Single-stage definitive laparoscopic management in mild acute biliary pancreatitis. AB - BACKGROUND: Population-based studies have shown that nearly one third of patients with acute biliary pancreatitis undergo endoscopic retrograde cholangiopancreatography (ERCP) before undergoing laparoscopic cholecystectomy (LC) (two-stage approach). The present study was designed to evaluate the safety of single-stage laparoscopic management to avoid preoperative ERCP. MATERIALS AND METHODS: Between June 1998 and June 2002, 35 female patients and 10 male patients (median age, 59 years) with uncomplicated mild acute biliary pancreatitis were studied prospectively and reviewed retrospectively. LC with fluoroscopic intraoperative cholangiography (IOC) or with fluoroscopic IOC and laparoscopic CBD exploration in cases of concomitant choledocholithiasis was performed as the definitive treatment (single-stage approach). Patients underwent surgery electively when symptoms had subsided and laboratory parameters had improved. RESULTS: LC alone was performed in 39 patients, and an additional laparoscopic CBD exploration was performed in the remaining six. In one patient, IOC yielded a false-positive result. CBD stones were detected in four cases, and debris in the CBD in one case, for an 11% incidence of concomitant choledocholithiasis. The conversion rate was zero, and single-stage laparoscopic treatment was successful in all cases. The overall morbidity rate was 4%. The 30-day postoperative mortality rate was zero. CONCLUSION: Although preoperative ERCP and sphincterotomy still have a role in complicated cases of mild acute biliary pancreatitis, laparoscopic single-stage definitive treatment is feasible and safe in uncomplicated cases of disease when local experience is available. PMID- 12737721 TI - Conversion rate in laparoscopic cholecystectomy: evolution from 1993 and current state. AB - The definition of difficult laparoscopic cholecystectomy (LC) is inconsistent. The aim of this study was to analyze the factors that make LC difficult to perform and determine ways to avoid conversion, based on our series. All patients who underwent LC or open cholecystectomy (OC) between January 1993 and December 2001 in our division of general surgery were the subject matter of this study. Preliminary decisions regarding LC or OC were avoided. Our experience (1993-2001) was based on 1360 consecutive elective LC procedures in 381 male and 979 female patients. The mean age of the patients at operation was 53 years (range, 17-84). The median operating time was 55 minutes (range, 35-180). The overall conversion rate was 1.8%. Indications for conversion included surgical difficulty during the laparoscopic procedure and anesthesia issues. The conversion rate has decreased to less than 1% in recent years. There were no mortalities, and the postoperative complication rates were low. The mean hospital stay of the patients was 2.6 days. In conclusion, based on our experience, we suggest limiting OC to patients with proven contraindications to LC (i.e., Mirizzi syndrome or systemic illness incompatible with pneumoperitoneum), attempting LC in all other cases, and considering cholecystostomy and delayed LC as an alternative to conversion during difficult LC. PMID- 12737720 TI - Selective application of laparoscopic intervention in the management of isolated bowel rupture in blunt abdominal trauma. AB - BACKGROUND: Recently, laparoscopy has been used as an effective diagnostic and therapeutic modality for traumatic injury and peritonitis. However, its clinical benefits are still under evaluation. The aim of this study was to assess the feasibility and safety of this technique, in comparison with traditional laparotomy, in the management of isolated bowel rupture. A further aim was to evaluate possible predictive clinical factors in the management of isolated bowel rupture. INDICATIONS: Patients with definite or suspected isolated gastrointestinal perforation who were hemodynamically stable underwent laparoscopic surgery for diagnosis and treatment. METHODS AND RESULTS: The traditional laparotomy control group (Group A, historical controls) included patients from a prior study. In this previous study, carried out between 1993 and 1997, 23 consecutive cases of traumatic gastrointestinal perforation were treated with traditional laparotomy. Retrospectively, 13 of these 23 cases would have been eligible for laparoscopic intervention. The laparoscopic group (group B) was made up of cases seen at our institution from 1998 to 2000. Of the 13 consecutive cases of traumatic gastrointestinal perforation, only one required immediate traditional laparotomy. Just one of the 12 cases of gastrointestinal perforation approached laparoscopically had to be converted to a traditional laparotomy. The following variables were evaluated in both groups: patient background, time from accident to onset of operation, operative time, blood loss, time to oral intake, incidence of peritoneal contamination, length of hospital stay, mortality, and intraoperative and postoperative complications. There were no statistical differences in age, gender ratio, and injury severity score (ISS). The mean operative times were 132.3 +/- 58.7 minutes and 143.6 +/- 27.3 minutes in group A and group B, respectively. The mean blood loss was 266.8 +/- 277.8 mL in group A and 57.6 +/- 57.1 mL in group B. The blood loss in group B was significantly lower (P =.0084) than that in group A. There were no significant differences in intraoperative and postoperative complications, hospital stay, and mortality between the two groups. CONCLUSIONS: Laparoscopic intervention offers better results in the management of patients with blunt abdominal trauma and isolated bowel rupture. PMID- 12737722 TI - Laparoscopic cholecystectomy: retrospective comparative evaluation of titanium versus absorbable clips. AB - We present a retrospective study of the use of titanium and absorbable clips during laparoscopic cholecystectomy. The aim was to determine any differences in outcome and costs. From January 1999 to February 2002, 690 patients who had successfully undergone a laparoscopic cholecystectomy were reviewed. According to the type of clip, we retrospectively identified two groups of patients: 199 in whom the surgeons had used absorbable clips (absorbable clip group, ACG) and 491 in whom the surgeons had used titanium nonabsorbable clips (titanium clip group, TCG). Data about demographics, operation, results, complications, and follow-up were collected and matched in the two groups. Demographics, concomitant surgery, and the American Society of Anesthesiologists (ASA) status were comparable between the two groups. Although the proportions of cases requiring urgent operation, intraoperative cholangiography, use of a fourth trocar, and use of drainage suction were similar, the difficulty score of the operation was lower (6.3 vs. 7.0, P =.03) and the operative time was shorter (44 vs. 61 minutes, P <.0001) in the ACG than in the TCG. Complications, hospital stay, and long-term results were satisfactory and comparable between the two groups. No correlation was found between clip type and the incidence of biliary tree injuries, bleeding, wound infection, or readmission. The cost of the two types of clips varied slightly (90 euros for each procedure). Despite the fact that absorbable clips are theoretically less likely to cause complications than metallic ones, we were not able to demonstrate any clinical advantage during laparoscopic cholecystectomy in this retrospective study. Furthermore, the results suggest that absorbable clips are preferred when the cholecystectomy presents fewer difficulties. PMID- 12737723 TI - Gasless video endoscopic transanal excision of rectal tumors incompletely removed by colonoscopic snare polypectomy. AB - BACKGROUND: This report describes an experience with gasless video transanal endoscopic microsurgery (VTEM) to excise rectal tumors previously incompletely removed with colonoscopic snare polypectomy. METHODS: Gasless VTEM involves a modification of transanal endoscopic microsurgery (TEM) that incorporates a standard laparoscopic video camera and requires no CO(2) insufflation system. Nineteen patients who had had a rectal tumor removed incompletely by colonoscopic polypectomy with a diathermy snare were enrolled in this prospective study. The patients included 14 men and 5 women whose median age was 63.5 (range, 49-83) years. The rectal tumors included 4 adenomas, 11 adenocarcinomas (Tis, 7; T1, 4), and 4 carcinoid tumors. The median distance from the tumor margin to the dentate line was 5.8 (range, 2.0-13.0) cm. RESULTS: All rectal lesions were successfully removed by gasless VTEM. No intraoperative complication occurred. The median operating time and blood loss were 40 (range, 15-145) minutes and 5 (range, 0 100) mL, respectively. The median maximal tumor diameter in 9 patients with residual tumors was 1.3 (range, 0.5-2.5) cm. There was no operative mortality. A postoperative complication (bleeding from a suture wound and transient incontinence) developed in 1 (5.3%) of the 19 patients. The median postoperative hospital stay was 5 (range, 2-10) days. Postoperative histology revealed a residual tumor in 10 (52.9%) of the 19 specimens. Complete excision of all tumors was confirmed histologically. During a median follow-up period of 59.5 (range, 12.3-94.9) months, no tumor recurred. CONCLUSIONS: Gasless VTEM is useful and minimally invasive for the local removal of rectal tumors incompletely resected by colonoscopic snare polypectomy. PMID- 12737724 TI - Isolated trans-hiatal colonic herniation. AB - Isolated herniation of the colon through congenital or traumatic diaphragmatic defects are well documented. However, trans-hiatal herniation of the colon in the absence of an intrathoracic stomach has been reported only once. A 67-year-old man presented with intragastric abdominal pain and a chest x-ray film documenting a posterior mediastinal air-fluid level. Computed tomography showed gastrointestinal contents within the thorax. The findings on an upper gastrointestinal film with small bowel follow-through were normal. Finally, a barium enema identified transverse colon within the thoracic cavity. At laparoscopy, the entire transverse colon was reduced with the hernia sac. The crural defect was repaired, and a Toupet fundoplication was performed. A gastropexy was also added. The patient was discharged on postoperative day 2 able to tolerate a regular diet, and he has been asymptomatic for 5 months. This defect most likely represents a congenital deformity of the diaphragm with intact posterior gastric attachments, including the posterior phrenoesophageal ligament. An intact gastric mesentery enabled isolated colonic herniation with retention of the stomach its normal anatomic position. An antireflux procedure was performed in addition to the crural repair because of the circumferential dissection of the esophagus. This article is the second report of an isolated trans-hiatal herniation of the colon and the first report of laparoscopic repair of this entity. PMID- 12737725 TI - Laparoscopic splenectomy for ruptured spleen: lessons learned from a case. AB - Splenic rupture is a frequent consequence of blunt abdominal trauma. When nonsurgical management is not indicated, splenic rupture is usually an emergency requiring surgery. Removal of the spleen (splenectomy) or alternative conservative procedures (splenorraphy, partial splenectomy, and hemostatic collagen application) are surgical treatment options. Recently, laparoscopic conservative techniques have been proposed. Laparoscopic splenectomy for a ruptured spleen has been reported only in a single case, in which a hand-assisted technique was used. We present the first reported case, to our knowledge, of successful removal of a ruptured spleen by means of a totally laparoscopic technique. Adequate technical instrumentation, such as the endostapler, harmonic scalpel, and red cell-saving machine, in addition to extensive experience with routine splenectomy, were essential factors to ensure a safe and effective procedure. PMID- 12737726 TI - Hand-assisted laparoscopic splenectomy for a huge splenic cyst: operative technique and case report. AB - We report the case of a huge splenic cyst that was successfully treated by hand assisted laparoscopic splenectomy. A 17-year-old girl with a chief complaint of left-sided abdominal pain was admitted to our department for investigation of a splenic tumor. Ultrasonography, computed tomography, and magnetic resonance imaging revealed a huge cystic lesion in the spleen measuring approximately 10 cm in diameter. Hand-assisted laparoscopic splenectomy was safely performed to diagnose and treat the splenic tumor. The histologic diagnosis was an epithelial cyst of the spleen with no atypical cells in the cyst wall. Hand-assisted laparoscopic splenectomy may be a good method of managing a huge splenic cyst that becomes symptomatic and potentially life-threatening through enlargement, rupture, and secondary infection. PMID- 12737727 TI - Laparoscopic resection of malignant gastric polyp-with a particular emphasis on localization by laparoscopic ultrasonography. AB - Adenomatous polyps of the stomach are rare and usually found on routine endoscopic examination. These polyps should be removed because of their potential for malignant transformation. Endoscopic polypectomy is the preferred treatment modality, but a laparotomy is required to resect large polyps. This study presents a case of a large gastric polyp containing carcinoma in situ that was successfully managed by laparoscopic gastrectomy. With the use of a novel technique, the tumor was localized by intraoperative laparoscopic ultrasonography. The patient had an excellent postoperative recovery, and follow up endoscopy at 24 months revealed no recurrent tumor. PMID- 12737728 TI - Video-assisted direct closure of bronchial fistula. AB - Video-assisted thoracoscopic surgery is a well-established method for managing persistent air leak in spontaneous pneumothorax. We describe a case of complicated spontaneous secondary pneumothorax in a patient with bullous emphysema that was treated by video-assisted manual suture of the bronchial fistula at the end of the right upper bronchus. PMID- 12737729 TI - Laparoscopic radical parametrectomy including paraaortic and pelvic lymphadenectomy in women after prior hysterectomy: three cases. AB - BACKGROUND: Radical abdominal surgery in patients who have previously undergone a hysterectomy is a surgical challenge. This type of surgery for invasive cervical cancer after a hysterectomy or for vaginal stump metastasis traditionally requires a major laparotomy; however, a minimal access approach is now being applied to this type of procedure. METHODS: A laparoscopic radical parametrectomy including a paraaortic and pelvic lymphadenectomy was performed on two patients with invasive cervical cancer diagnosed after a simple hysterectomy and one patient with recurrent endometrial cancer in the vaginal stump. RESULTS: All three patients had an excellent clinical outcome and made a rapid recovery, even though two cases involved a bladder laceration. CONCLUSION: A laparoscopic radical parametrectomy including a paraaortic and pelvic lymphadenectomy is a viable technique for women with invasive cervical cancer or recurrent endometrial vaginal cancer after a prior hysterectomy. PMID- 12737730 TI - Laparoscopic drainage of lymphocele after kidney transplant. AB - Lymphocele can develop after renal transplantation. Surgical internal drainage to the abdominal cavity through a standard laparotomy is indicated for symptomatic persistent lesions. Internal drainage can be performed laparoscopically. In this report, we describe our laparoscopic surgical technique for drainage and analyze our results in five patients with lymphoceles that developed after kidney transplantation. All the patients were male, with a mean age of 29 +/- 10 years. The volume of fluid in the lymphoceles ranged from 500 to 1000 mL. Percutaneous drainage was selected as the initial treatment without success. A laparoscopic peritoneal window was created in all patients under intraoperative ultrasonographic guidance. The mean operative time was 90 minutes. In all patients, the fluid collections resolved after laparoscopic internal drainage without complications. During a mean follow-up of 3 +/- 2 years, all patients remained asymptomatic, and no additional fluid collections were identified. PMID- 12737731 TI - Laparoscopic implantation of swan neck presternal peritoneal dialysis catheters. AB - The swan neck presternal peritoneal dialysis catheter provides an alternative location from which a catheter can exit the skin when an abdominal site is not suitable. The exit site is in the upper anterior chest. The presternal catheter was designed for patients who would not ordinarily be considered for peritoneal dialysis because of body habitus, presence of stomas, or urinary-fecal incontinence. In its original design, the catheter can be implanted only by open dissection. We present a modification of the catheter system and describe an operative technique in which the catheter can be inserted laparoscopically. Eight patients underwent laparoscopic implantation of presternal catheters by this technique. During follow-up ranging from 2.9 to 12.4 months (average, 5.3 months), no instances of catheter dysfunction, leakage around the cannula, hernia, or loss secondary to infection occurred. We foresee greater use of the swan neck presternal peritoneal dialysis catheter once surgeons become more familiar with its ease of insertion and if the device can be made available as a single-package system compatible with laparoscopic implantation. PMID- 12737732 TI - Planning against biological terrorism: lessons from outbreak investigations. AB - We examined outbreak investigations conducted around the world from 1988 to 1999 by the Centers for Disease Control and Prevention's Epidemic Intelligence Service. In 44 (4.0%) of 1,099 investigations, identified causative agents had bioterrorism potential. In six investigations, intentional use of infectious agents was considered. Healthcare providers reported 270 (24.6%) outbreaks and infection control practitioners reported 129 (11.7%); together they reported 399 (36.3%) of the outbreaks. Health departments reported 335 (30.5%) outbreaks. For six outbreaks in which bioterrorism or intentional contamination was possible, reporting was delayed for up to 26 days. We confirmed that the most critical component for bioterrorism outbreak detection and reporting is the frontline healthcare profession and the local health departments. Bioterrorism preparedness should emphasize education and support of this frontline as well as methods to shorten the time between outbreak and reporting. PMID- 12737733 TI - Endemic gastrointestinal anthrax in 1960s Lebanon: clinical manifestations and surgical findings. AB - Anthrax is an ancient disease caused by the gram-positive Bacillus anthracis; recently, it has gained much attention because of its potential use in biologic warfare. Anthrax infection occurs in three forms: cutaneous, inhalational, and gastrointestinal. The last type results from ingestion of poorly cooked contaminated meat. Intestinal anthrax was widely known in Lebanon in the 1960s, when a series of >100 cases were observed in the Bekaa Valley. We describe some of these cases, introduce the concept of the surgical management of advanced intestinal anthrax, and describe some of the approaches for treatment. PMID- 12737734 TI - Vero cytotoxin-producing Escherichia coli O157 gastroenteritis in farm visitors, North Wales. AB - An outbreak of Vero cytotoxin-producing Escherichia coli O157 (VTEC O157) gastroenteritis in visitors to an open farm in North Wales resulted in 17 primary and 7 secondary cases of illness. E. coli O157 Vero cytotoxin type 2, phage type 2 was isolated from 23 human cases and environmental animal fecal samples. A case control study of 16 primary case-patients and 36 controls (all children) showed a significant association with attendance on the 2nd day of a festival, eating ice cream or cotton candy (candy floss), and contact with cows or goats. On multivariable analysis, only the association between illness and ice cream (odds ratio [OR]=11.99, 95% confidence interval [CI] 1.04 to 137.76) and cotton candy (OR=51.90, 95% CI 2.77 to 970.67) remained significant. In addition to supervised handwashing, we recommend that foods on open farms only be eaten in dedicated clean areas and that sticky foods be discouraged. PMID- 12737735 TI - Pandemic influenza and healthcare demand in the Netherlands: scenario analysis. AB - In accordance with World Health Organization guidelines, the Dutch Ministry of Health, Welfare and Sports designed a national plan to minimize effects of pandemic influenza. Within the scope of the Dutch pandemic preparedness plan, we were asked to estimate the magnitude of the problem in terms of the number of hospitalizations and deaths during an influenza pandemic. Using scenario analysis, we also examined the potential effects of intervention options. We describe and compare the scenarios developed to understand the potential impact of a pandemic (i.e., illness, hospitalizations, deaths), various interventions, and critical model parameters. Scenario analysis is a helpful tool for making policy decisions about the design and planning of outbreak control management on a national, regional, or local level. PMID- 12737736 TI - Estimating the incidence of typhoid fever and other febrile illnesses in developing countries. AB - To measure the incidence of typhoid fever and other febrile illnesses in Bilbeis District, Egypt, we conducted a household survey to determine patterns of health seeking among persons with fever. Then we established surveillance for 4 months among a representative sample of health providers who saw febrile patients. Health providers collected epidemiologic information and blood (for culture and serologic testing) from eligible patients. After adjusting for the provider sampling scheme, test sensitivity, and seasonality, we estimated that the incidence of typhoid fever was 13/100,000 persons per year, and the incidence of brucellosis was 18/100,000 persons per year in the district. This surveillance tool could have wide applications for surveillance for febrile illness in developing countries. PMID- 12737737 TI - Human milk secretory antibodies against attaching and effacing Escherichia coli antigens. AB - Secretory immunoglobulin A (sIgA) is a primary factor responsible for preventing attachment of enteropathogens to gut epithelium in breastfeeding infants. We compared the frequency of sIgA to major surface antigens of enterohemorrhagic Escherichia coli (EHEC) in milk of 123 women from the United States and Mexico to determine whether regional differences existed in the frequency of antibodies to these surface antigens. In both groups of women, milk commonly has sIgA against various EHEC lipopolysaccharides, EspA, EspB, intimin, and less frequently against Shiga toxin. The study suggests that persons living in the United States are exposed to attaching/effacing enteropathogens more frequently than is generally assumed. The low frequency of antibodies to Stx1 (in 12% of Mexican and in 22% of U.S. samples) suggests that the rare appearance of hemolytic uremic syndrome in adults is not due to neutralization of toxin at the gut level. Only anti-EspA is found in most milk samples from both populations of women. EspA may represent a useful target for an immunization strategy to prevent EHEC disease in humans. PMID- 12737738 TI - Aeromonas spp. and traveler's diarrhea: clinical features and antimicrobial resistance. AB - Traveler's diarrhea is the most common health problem of international travelers. We determined the prevalence of Aeromonas spp. associated with traveler's diarrhea and analyzed the geographic distribution, clinical features, and antimicrobial susceptibility. Aeromonas spp. were isolated as a cause of traveler's diarrhea in 18 (2%) of 863 patients. A. veronii biotype sobria was isolated in nine patients, A. caviae in seven patients, and A. jandaei and A. hydrophila in one patient each. Aeromonas spp. were isolated with a similar prevalence in Africa, Latin America, and Asia. Watery and persistent diarrhea, fever, and abdominal cramps were common complaints. All strains were resistant to ampicillin; showed variable resistance to chloramphenicol, tetracycline, and cotrimoxazole; and were susceptible to cefotaxime, ciprofloxacin, and nalidixic acid. The persistence of symptoms made antimicrobial treatment necessary. PMID- 12737739 TI - Endemic, notifiable bioterrorism-related diseases, United States, 1992-1999. AB - Little information is available in the United States regarding the incidence and distribution of diseases caused by critical microbiologic agents with the potential for use in acts of terrorism. We describe disease-specific, demographic, geographic, and seasonal distribution of selected bioterrorism related conditions (anthrax, botulism, brucellosis, cholera, plague, tularemia, and viral encephalitides) reported to the National Notifiable Diseases Surveillance System in 1992 to 1999. Tularemia and brucellosis were the most frequently reported diseases. Anthrax, plague, western equine encephalitis, and eastern equine encephalitis were rare. Higher incidence rates for cholera and plague were noted in the western United States and for tularemia in the central United States. Overall, the incidence of conditions caused by these critical agents in the United States is low. Individual case reports should be considered sentinel events. For potential bioterrorism-related conditions that are endemic and have low incidence, the use of nontraditional surveillance methods and complementary data sources may enhance our ability to rapidly detect changes in disease incidence. PMID- 12737741 TI - Seasonal patterns of invasive pneumococcal disease. AB - Pneumococcal infections increase each winter, a phenomenon that has not been well explained. We conducted population-based active surveillance for all cases of invasive pneumococcal disease in seven states; plotted annualized weekly rates by geographic location, age, and latitude; and assessed correlations by time-series analysis. In all geographic areas, invasive pneumococcal disease exhibited a distinct winter seasonality, including an increase among children in the fall preceding that for adults and a sharp spike in incidence among adults each year between December 24 and January 7. Pneumococcal disease correlated inversely with temperature (r -0.82 with a 1-week lag; p<0.0001), but paradoxically the coldest states had the lowest rates, and no threshold temperature could be identified. The pattern of disease correlated directly with the sinusoidal variations in photoperiod (r +0.85 with a 5-week lag; p<0.0001). Seemingly unrelated seasonal phenomena were also somewhat correlated. The reproducible seasonal patterns in varied geographic locations are consistent with the hypothesis that nationwide seasonal changes such as photoperiod-dependent variation in host susceptibility may underlie pneumococcal seasonality, but caution is indicated in assigning causality as a result of such correlations. PMID- 12737740 TI - Global illness and deaths caused by rotavirus disease in children. AB - To estimate the global illness and deaths caused by rotavirus disease, we reviewed studies published from 1986 to 2000 on deaths caused by diarrhea and on rotavirus infections in children. We assessed rotavirus-associated illness in three clinical settings (mild cases requiring home care alone, moderate cases requiring a clinic visit, and severe cases requiring hospitalization) and death rates in countries in different World Bank income groups. Each year, rotavirus causes approximately 111 million episodes of gastroenteritis requiring only home care, 25 million clinic visits, 2 million hospitalizations, and 352,000-592,000 deaths (median, 440,000 deaths) in children <5 years of age. By age 5, nearly every child will have an episode of rotavirus gastroenteritis, 1 in 5 will visit a clinic, 1 in 65 will be hospitalized, and approximately 1 in 293 will die. Children in the poorest countries account for 82% of rotavirus deaths. The tremendous incidence of rotavirus disease underscores the urgent need for interventions, such as vaccines, particularly to prevent childhood deaths in developing nations. PMID- 12737742 TI - Entamoeba moshkovskii infections in children, Bangladesh. AB - Entamoeba moshkovskii cysts are morphologically indistinguishable from those of the disease-causing species E. histolytica and the nonpathogenic E. dispar. Although sporadic cases of human infection with E. moshkovskii have been reported, the organism is considered primarily a free-living amoeba. No simple molecular detection tool is available for diagnosing E. moshkovskii infections. We used polymerase chain reaction (PCR) to detect E. moshkovskii directly in stool. We tested 109 stool specimens from preschool children in Bangladesh by PCR; 17 were positive for E. histolytica (15.6%) and 39 were positive for E. dispar (35.8%). In addition, we found that 23 (21.1%) were positive for E. moshkovskii infection, and 17 (73.9%) of these also carried E. histolytica or E. dispar. The high association of E. moshkovskii with E. histolytica and E. dispar may have obscured its identification in previous studies. The high prevalence found in this study suggests that humans may be a true host for this amoeba. PMID- 12737743 TI - Variant Salmonella genomic island 1 antibiotic resistance gene cluster in Salmonella enterica serovar Albany. AB - Salmonella genomic island 1 (SGI1) contains an antibiotic resistance gene cluster and has been previously identified in multidrug-resistant Salmonella enterica serovars Typhimurium DT104, Agona, and Paratyphi B. We identified a variant SGI1 antibiotic-resistance gene cluster in a multidrug-resistant strain of S. enterica serovar Albany isolated from food fish from Thailand and imported to France. In this strain, the streptomycin resistance aadA2 gene cassette in one of the SGI1 integrons was replaced by a dfrA1 gene cassette, conferring resistance to trimethoprim and an open reading frame of unknown function. Thus, this serovar Albany strain represents the fourth S. enterica serovar in which SGI1 has been identified and the first SGI1 example where gene cassette replacement took place in one of its integron structures. The antibiotic resistance gene cluster of serovar Albany strain 7205.00 constitutes a new SGI1 variant; we propose a name of SGI1-F. PMID- 12737744 TI - Emerging rickettsioses of the Thai-Myanmar border. AB - To investigate the presence of rickettsioses in rural residents of the central Thai-Myanmar border, we tested the blood of 46 patients with fever. Four patients had murine typhus, three patients had scrub typhus, and eight patients had spotted fever group rickettsioses, including the first case of Rickettsia felis infection reported in Asia. PMID- 12737746 TI - Chronic wasting disease in free-ranging Wisconsin White-tailed Deer. AB - Three White-tailed Deer shot within 5 km during the 2001 hunting season in Wisconsin tested positive for chronic wasting disease, a prion disease of cervids. Subsequent sampling within 18 km showed a 3% prevalence (n=476). This discovery represents an important range extension for chronic wasting disease into the eastern United States. PMID- 12737747 TI - Human metapneumovirus and community-acquired respiratory illness in children. PMID- 12737745 TI - Eliminating trachoma in areas with limited disease. AB - The common wisdom is that a trachoma program cannot eliminate ocular chlamydia from a community, just reduce infection to a level where blindness would be minimal. We describe the success of multiple mass antibiotic treatments, demonstrating that complete elimination of infection may be an attainable goal in an area with modest disease. PMID- 12737748 TI - Puumala virus infection with acute disseminated encephalomyelitis and multiorgan failure. PMID- 12737749 TI - Pregnancy and asymptomatic carriage of Pneumocystis jiroveci. PMID- 12737750 TI - First evidence of Aedes albopictus (Skuse) in southern Chiapas, Mexico. PMID- 12737751 TI - Virus isolation and "acute" West Nile virus encephalitis (response to Huang et al.). PMID- 12737753 TI - Malaria prophylaxis: setting the scene. PMID- 12737754 TI - Atovaquone/proguanil: the need for family protection. AB - An increasing number of families, including children and the elderly, are seeking more adventurous travel in exotic parts of the world. Holiday destinations now include once-remote regions such as subSaharan Africa and New Guinea. This increase in visits to tropical and subtropical regions, combined with widespread chloroquine-resistant malaria, now places millions of Western travelers at risk of infection annually. At least 30,000 travelers from industrialized countries are reported to contract malaria each year and approximately 1 in 100 travelers who acquire Plasmodium falciparum malaria will die. PMID- 12737756 TI - Post-marketing surveillance: adverse events during long-term use of atovaquone/proguanil for travelers to malaria-endemic countries. AB - BACKGROUND: Atovaquone/proguanil in Europe is only licensed for 4 weeks of travel. Data on its long-term tolerability in nonimmune travelers are scarce. METHODS: We prospectively studied adverse reactions reported by long-term travelers using atovaquone/proguanil among a population intolerant to mefloquine. The average length of atovaquone/proguanil use was 9 weeks (4.5 to 34 weeks). Adverse events were recorded on a regular questionnaire. Travelers rated complaints as: (1) mild, not interfering with their daily activities; (2) moderate, causing interference, such as canceling a trip or being confined to the hotel; or (3) severe, causing a visit to a doctor or clinic. Hospital admission was classified as category 3 and specified. We then compared our data with those on adverse reactions from 2 large multicenter studies of atovaquone/proguanil in nonimmune travelers. RESULTS: One hundred and fifty-four subjects used atovaquone/proguanil for a total of 1538 weeks. Diarrhea was the most common ailment (18%). Further complaints were abdominal pain (mild 4%, moderate 5%, severe 2%), headache (mild 4%, moderate 4%, severe 1%), dizziness (mild 3%, moderate 1%, severe 1%), and insomnia (mild 6%, moderate 0%, severe 0%). Two subjects of 154 (1%) discontinued prophylaxis, both due to diarrhea. Nobody was admitted to hospital. No case of malaria was reported. Comparison with data from previous, larger atovaquone/proguanil studies shows that the adverse events reported by the long-term atovaquone/proguanil users are no different in type and frequency of occurrence to those travel-related health problems normally encountered in the Tropics. CONCLUSIONS: Atovaquone/proguanil was well tolerated by these long-term travelers. Long-term users of atovaquone/ proguanil antimalarial prophylaxis suffer no more ailments than normally occur to travelers in tropical regions. PMID- 12737755 TI - The safety of atovaquone/proguanil in long-term malaria prophylaxis of nonimmune adults. AB - BACKGROUND: Data on the long-term safety of atovaquone/proguanil in nonimmune travelers are limited. METHODS: An open-label study, involving 300 Danish soldiers stationed in Eritrea for 6 months was initiated. The subjects self reported their symptoms on a post-travel questionnaire. The study compared the symptoms of compliers and noncompliers. RESULTS: No serious adverse events occurred. Diarrhea, stomach pain, headache, cough, and loss of appetite were the most common symptoms reported. No case of Plasmodium falciparum malaria occurred. CONCLUSIONS: Atovaquone/proguanil was safe and well tolerated in this group of long-term nonimmune travelers. PMID- 12737759 TI - Breastfeeding reduces pain in neonates. PMID- 12737760 TI - Zinc nasal gel effective for the common cold. PMID- 12737761 TI - Use of sputum eosinophil count decreases asthma exacerbations. PMID- 12737763 TI - Low-dose tricyclics effective for depression. PMID- 12737762 TI - Ultrasonography helpful in diagnosing developmental hip dysplasia. PMID- 12737764 TI - What is a reasonable interval for retinopathy screening in patients with diabetes? PMID- 12737765 TI - Nebulized 3% saline effective for viral bronchiolitis. PMID- 12737766 TI - Optimal digoxin range for men is 0.5 to 0.8 ng/mL. PMID- 12737767 TI - Transdermal progesterone ineffective for menopausal symptoms. PMID- 12737768 TI - What is the best NSAID regimen for arthritis patients with bleeding ulcer? PMID- 12737769 TI - Office evaluation and treatment of hemorrhoids. PMID- 12737770 TI - A terrible itch. PMID- 12737771 TI - Evidence or bias? PMID- 12737772 TI - Asthma: resource use and costs for inhaled corticosteroid vs leukotriene modifier treatment--a meta-analysis. AB - OBJECTIVE: To compare the effects of inhaled corticosteroid treatment with leukotriene modifier treatment on medical resource use and costs for asthma patients. STUDY DESIGN: Meta-analysis combining results from published and unpublished studies. DATA SOURCES: Studies were identified from the MEDLINE and EMBASE databases and the GlaxoSmithKline internal database study registers. Two independent reviewers evaluated the identified studies; studies meeting specified inclusion criteria were abstracted and summarized by meta-analysis with a random effects model. OUTCOMES MEASURED: Hospitalization rate, emergency department visit rate, emergency department costs, drug costs, total asthma-related costs, and total medical care costs. RESULTS: Patients taking inhaled corticosteroids had: a significantly lower annual rate of hospitalization than those taking leukotriene modifiers (2.2% vs 4.3%, respectively; P<.05); a greater decline in hospitalization rate (before vs after therapy initiation) than those taking leukotriene modifiers (decline of 2.4% vs 0.55%; P<.01); a lower annual rate of emergency department visits than those taking leukotriene modifiers (6.2% vs 7.7%; P<.005); lower total asthma-related medical costs than those taking leukotriene modifiers (P<.05) and a 17% reduction in overall total medical care costs (P not significant). CONCLUSIONS: Patients with asthma treated with inhaled corticosteroids have significantly fewer asthma-related hospitalizations and emergency department visits and lower total asthma-related health care costs than patients treated with leukotriene modifiers. These meta-analysis findings are consistent with results from randomized controlled trials showing improvements in lung function for patients taking inhaled corticosteroids as opposed to leukotriene modifiers. PMID- 12737774 TI - Clinical inquiries. Does increasing methylphenidate dose aid symptom control in ADHD? PMID- 12737773 TI - Clinical inquiries. What is the most effective beta-blocker for heart failure? PMID- 12737776 TI - Clinical inquiries. How should we manage infants at risk for group B streptococcal disease? PMID- 12737775 TI - Clinical inquiries. Are tympanostomy tubes indicated for recurrent acute otitis media? PMID- 12737777 TI - The art of medicine after retirement. PMID- 12737778 TI - [Changes in myocardial function and perfusion after acute myocardial infarction]. PMID- 12737779 TI - [Smoking and coronary disease: smoking cessation is the best drug]. PMID- 12737781 TI - [Smoking cessation and risk of myocardial reinfarction in coronary patients: a nested case-control study]. AB - INTRODUCTION AND OBJECTIVES: Smoking cessation reduces mortality in coronary patients. The aim of this study was to estimate association measures between the risk of occurrence of fatal or non-fatal reinfarction in patients who either continue to smoke or stop after a first infarction and are treated with secondary prevention measures. PATIENTS AND METHOD: The study was a case-control (1:1) design nested in a cohort of 985 coronary patients under the age of 76 years who were not treated with invasive procedures and survived more than 6 months after the first acute myocardial infarction. Cases were all patients who suffered reinfarction (n = 137) between 1997 and 2000. A control patient was matched with each case by gender, age, hospital, interviewer, and the secondary prevention timeframe. RESULTS: Patients who smoke after the first acute myocardial infarction had an Odds ratio (OR) of 2.83 (95% CI, 1.47-5.47) for a new acute myocardial infarction. Adjustment for lifestyle, drug treatment, and risk factors (family history of coronary disease, high blood pressure, hypercholesterolemia, and diabetes mellitus) did not change the OR (2.80 [95% CI, 1.35-5.80]). Patients who quit smoking had an adjusted OR of 0.90 (95% CI, 0.47-1.71) compared with non smokers before the first acute myocardial infarction. Continued smoking had an adjusted OR of 2.90 (95% CI, 1.35-6.20) compared to quitting after the first acute myocardial infarction. CONCLUSION: The risk of acute myocardial infarctions is three times higher in patients who continue to smoke after an acute coronary event compared with patients who quit. The risk of reinfarction in patients who stop smoking is similar to the risk of non-smokers before the first infarction. PMID- 12737780 TI - [Evolution of systolic function and myocardial perfusion, evaluated by gated SPECT, in the first year after acute myocardial infarction]. AB - INTRODUCTION AND OBJECTIVES: Systolic function and myocardial perfusion are evaluated before hospital discharge and can change during follow-up. The purpose of this study was to evaluate these parameters by gated-SPECT in the first year after acute myocardial infarction. PATIENTS AND METHOD: We studied 74 consecutive patients with a first uncomplicated acute myocardial infarction (49 infero lateral and 25 anterior) by stress-rest 99mTc-tetrofosmin and rest-gated-SPECT before hospital discharge (6-8 days after admission) and one year after myocardial infarction. RESULTS: The ejection fraction (EF) increased > 5% in 51% of infero-lateral infarcts and 28% of non-revascularized anterior infarcts. EF increased significantly (48.4 8% to 54.6 8.7%; p < 0.0001, mean difference: 6.2; 95% IC, 2.8-9.5) and systolic volume decreased (51.3 19.2 ml to 44.3 19.4 ml; p = 0.001; mean diff.: 7.67; 95% IC, 1.5-13.8) in infero-lateral infarctions. The rest perfusion index in the necrotic region improved (2.3 0.57 to 2.17 0.58; p = 0.004; mean diff.: 0.18; 95% IC, 0.003-0.36) in infero-lateral infarcts and the ischemia index remained unchanged between the first and second studies. CONCLUSIONS: Left ventricular systolic function can change during the first year of evolution, a significant improvement being seen in infero-lateral infarctions. The ejection fraction increased > 5% in half of these patients, as opposed to only a quarter of anterior infarctions. This improvement was associated to increased myocardial perfusion at rest. PMID- 12737782 TI - [Coronary risk factors in adolescence. The FRICELA study]. AB - AIMS: In a grou of adolescents, to examine the prevalence of behaviors and risk factors that predispose to coronary heart disease and ascertain their associations. METHODS: A population of 2,599 adolescents, age 12 to 19 years, both sexes, was studied. The study sites were 30 public or private medical centers located in the Federal District and 12 provinces of Argentina. RESULTS: Teenagers with hypercholesterolemia had a greater body mass index, higher incidence of hypertension, and more frequent sedentary lifestyle. There was a positive correlation between hypertension, body mass index, and family history of hypertension, and a negative correlation with physical exercise. Obesity in the young correlated with obesity in their parents. At the age of 19 years, 42% consumed alcohol and 28% smoked. The association between smoking and alcohol consumption was very high (p < 0.0001). The smoking habits of parents strongly influenced their children. CONCLUSIONS: Early inculcation of a healthy lifestyle, consisting of regular physical exrcise, a healthy diet, and no smoking, could be the most important behavorial and risk factor interventions in the effort to curtail atherosclerosis in adults. PMID- 12737783 TI - [Late recovery of coronary flow reserve in patients successfully treated with a percutaneous procedure]. AB - INTRODUCTION AND OBJECTIVES: Coronary angiograms are of limited value for the assessment of the results of percutaneous interventions. Intracoronary Doppler studies have been used to overcome these difficulties. The achievement of a coronary flow reserve (CFR) > 2-2.5 after the procedure is considered a good result and further optimization is generally not required. However, coronary flow reserve may not recover immediately, despite optimal procedural results. The aim of this study is to assess the temporal course of the recovery of coronary flow reserve after successful revascularization. PATIENTS AND METHOD: We studied 34 patients with coronary heart disease who were successfully treated by balloon angioplasty (n = 8) or stent implantation (n = 26). In all patients, serial observations were made by quantitative angiography and intracoronary Doppler (0.014 = flow-wire). Patients were studied: a) before treatment; b) immediately after, and c) 8 3 months later. RESULTS: The baseline coronary flow reserve was 1.3 0.4 and increased to 2.4 0.8 after the procedure (p < 0.01). At 8 months follow-up there was a significant increase (3 0.8; p < 0.01). This late improvement in coronary flow reserve was associated with a decline in average peak velocity at follow-up. Patients with impaired CFR immediately after treatment had a greater increase in CFR during followup than those with CFR > 2 after treatment (1.4 0.9 vs 0.4 0.6; p < 0.01). CONCLUSIONS: After a successful coronary intervention, CFR increases immediately, but some patients may experience additional improvement during follow-up. This increase was greater in patients who showed less improvement in coronary flow reserve immediately after treatment. Our findings suggest that the use of Doppler parameters in the immediate assessment of percutaneous coronary intervention results have limitations. PMID- 12737784 TI - [Late T-lymphocyte and monocyte activation in coronary restenosis. Evidence for a persistent inflammatory/immune mechanism?]. AB - AIMS: This study was made to determine if restenosis after percutaneous coronary angioplasty is associated with acute or chronic inflammatory/immunologic activity, and explored possible relationships with latent infection. PATIENTS AND METHOD: Forty-six consecutive patients underwent elective PTCA and 6 months of angiographic follow-up. Peripheral venous blood samples were obtained at baseline, 24-48 h, and 4-6 months post-intervention. Flow-cytometric methods were used to measure early and late circulating leukocyte activation status. Il-6 and TNF-alpha cytokines, and Il-2 soluble receptor concentrations were determined in all plasma samples. Chlamydia pneumoniae and Cytomegalovirus antibody assays were performed to detect infectious disease. RESULTS: Angiographic coronary stenosis developed in 27 out of 46 patients. At 6 months of follow-up, these patients showed a significant increase in circulating cytotoxic T-lymphocytes CD3+/CD56+ (18.8 7.1 vs 6.12 2.7%; p = 0.005) and activated monocytes (CD11b: 1,383 624 vs 990 484 MFI, p = 0.025; CD64: 76.0 28.7 vs 56.7 21.8 MFI; p = 0.014), with no apparent relation to increased cytokines or latent infectious disease. CONCLUSIONS: Restenosis appears to be associated to inflammatory and immunological activity that persists 6 months after coronary intervention. No relationship was found with the infections studied. The presence of inflammatory activity 4-6 months after PTCA suggess that pharmacological therapeutic interventions to prevent restenosis should be maintained for months. PMID- 12737785 TI - [Coronary ectasia: prevalence, and clinical and angiographic characteristics]. AB - INTRODUCTION AND OBJECTIVES: Coronary ectasia is characterized by the presence of diffuse dilation of the coronary vessels and is detected in 0.3-5.3% of angiographic studies. Our objective was to evaluate the prevalence of this condition, to analyze its clinical and angiographic characteristics, and to compare patients with ectasia and patients without it. PATIENTS AND METHOD: Coronary angiography was performed in 4.332 patients from October 1998 to June 2001. This population was divided in two groups, patients with and patients without ectasia and patients without ectasia. Angiographic and clinical variables were compared in these groups. RESULTS: The prevalence of ectasia was 3.39%. Most patients with ectasia (77.6%) had coronary stenosis. Ectasia affected a single vessel in 49.7%, most frequently the right coronary artery (132 patients), which also showed the greatest dilation. Most patients with ectasia were men (91.2%), smokers (56.5%), and younger than patients without ectasia (60.8 11.7 vs. 63.3 10.7 years; p = 0.01). They also had a lower prevalence of diabetes (22.4%) and previous revascularization procedures (8.2% angioplasty and 1.4% surgical revascularization).Logistical regression analysis showed that only male sex was associated to the presence of ectasia (OR = 3.33; 95% CI, 1.81-6.13) and that only diabetes was independently associated with absence of ectasia (OR = 0.65; 95% CI, 0.43-0.98). CONCLUSIONS: The prevalence of coronary ectasia in patients who underwent angiography was 3.4%. Coronary ectasia was prevalent in males and associated to the classic cardiovascular risk factors, except diabetes, a pathology that was less frequent than usual. PMID- 12737786 TI - [Comparison of echocardiographic studies made with new portable devices to conventional studies]. AB - OBJECTIVES: The latest development in echocardiography is the hand-held ultrasound device. Previous studies have shown that portable ultrasound devices detect major cardiovascular pathology better than the physical examination, but their diagnostic accuracy is still not known. The purpose of this study was to compare the results of examinations made with portable devices to those obtained with higher-scale platforms. PATIENTS AND METHOD: 211 consecutive unselected patients were included in the study. Patients were randomly studied with a portable device and a standard platform (considered the gold standard for comparison) by cardiologists experienced in echocardiography. Parameters of cardiac morphology and function, and valvular regurgitation were compared and analyzed using the McNemar paired test. Differences of more than one grade were considered major differences. RESULTS: The subjective assessment of the studies made with the portable device was significantly worse. The correlation between estimates of left ventricular function (differences not statistically significant) was adequate, but significant differences were detected in the evaluation of left atrial enlargement, left ventricular hypertrophy, aortic root enlargement, and mitral and tricuspid regurgitation. CONCLUSIONS: Hand-held cardiac ultrasound devices do not satisfy criteria for a complete echocardiographic study. They provide accurate information about ventricular function but fail to adequately measure cardiac chambers or assess valve function. PMID- 12737787 TI - [2003 update of the Guidelines of the Spanish Society of Cardiology on High Blood Pressure]. AB - Since publication of the Spanish Society of Cardiology Clinical Practice Guidelines on High Blood Pressure in January 2000, a new body of scientific evidence has been obtained that needs to be taken into account in clinical practice. A complete clinical evaluation by assessment of the global cardiovascular risk score should be done in patients with hypertension. In this connection, ECG findings and urine albumin excretion are of particular value. Up to now, the results of most important clinical trials indicate that the aim should be to normalize blood pressure, with stricter control in patients at higher risk (diabetes, target organ damage or left ventricular hypertrophy). Antihypertensive therapy should be selected on an individual basis, taking in account that patients with certain associated pathologies will benefit more from particular groups of drugs. Those with diabetes or left ventricular hypertrophy seem to benefit from pharmacological block of the renin-angiotensin system, and patients with heart failure from combined therapy with ACE inhibitors plus beta blockers. PMID- 12737788 TI - [Imaging diagnosis in acute aortic syndromes]. AB - Acute aortic dissection is a clinical emergency. Its prognosis is related mainly with prompt and accurate diagnosis, as well as rapid treatment. In this paper we review the importance of different imaging techniques in the diagnosis of patients with acute aortic syndrome. Aortic dissection, intramural haematoma, and penetrating aortic ulcer are discussed. PMID- 12737789 TI - [Angio-MRI diagnosis of a post-surgical aortic aneurysm]. PMID- 12737790 TI - [Management by hypothermia of junctional ectopic tachycardia appearing after pediatric heart surgery]. AB - Junctional ectopic tachycardia (JET) is a complication of the repair of congenital cardiac malformations that responds poorly to conventional treatment. We report our experience with the use of moderate hypothermia in its management. Twelve infants with postoperative JET treated with hypothermia were reviewed. The mean interval between the diagnosis of JET and initiation of hypothermia was 1.5 0.5 hours. In the first 24 hours of hypothermia, central temperature and heart rate decreased significantly. Arterial pressure and diuresis tended to increase and central venous pressure tended to decrease. No direct adverse events occurred. All the patients but one survived and are alive and free of neurological deficits after 15 12 months. PMID- 12737791 TI - [Coronary endarterectomy and bypass grafting without cardiopulmonary bypass]. AB - We prospectively analyzed patients who underwent simultaneous off-pump coronary artery bypass grafting and endarterectomy between March 1, 1997 and February 28, 2002. The incidence of perioperative myocardial infarction, need for inotropic support, morbidity, long-time functional class, and mortality were evaluated. Nine endarterectomies were performed in eight patients, more frequently in the right coronary artery. Dopamine was used in four patients. One perioperative myocardial infarction (12.5%) occurred. No deaths occurred and all patients are now functional class I. Tests for ischemia have been negative in all patients.Coronary endarterectomy is an alternative procedure that has little morbidity and enables complete myocardial revascularization without cardiopulmonary bypass. PMID- 12737792 TI - [Incessant atrial tachycardia and a cerebral tumor]. AB - We report the case of an asymptomatic 78-year-old woman in whom incessant atrial tachycardia refractory to many pharmacological treatments appeared. Months after the appearance of atrial tachycardia, clinical neurological symptoms of a cerebral cystic tumor appeared. Treatment of the tumor resolved the rhythm disorder. PMID- 12737793 TI - [Protein-losing enteropathy after Fontan's operation] PMID- 12737794 TI - Nuclear receptors orchestrate detoxification pathways. AB - Two nuclear receptors of xenobiotic drugs, PXR and CAR, are central regulators of detoxification enzymes. New studies extend the role of these receptors to a natural detoxification process. They coordinate induction of proteins for storage, glucuronidation, and canalicular transport of bilirubin. PMID- 12737795 TI - Caspase activation finds fertile ground. AB - Cytochrome c is a critical regulator of apoptosome assembly, caspase activation, and programmed cell death. Recent work demonstrates that cytochrome c and caspases function in Drosophila sperm cell differentiation and indicates that caspase activity can be regulated in a subcellular manner in cells that live. PMID- 12737796 TI - Never fat or gaunt. AB - Bacteria stringently regulate the synthesis of their membrane phospholipids, but the responsible regulatory mechanisms are incompletely understood. In this issue of Developmental Cell, a study reports negative regulation of the transcription of several genes of fatty acid and phospholipid synthesis plus identification of the regulatory protein. PMID- 12737797 TI - Welcome to syndetome: a new somitic compartment. AB - Virtually nothing was known about the embryonic origin of tendons, until a recent paper by Brent and colleagues in which they track the origin of tendon progenitors of the body axis and reveal the molecular events and tissue interactions leading to their commitment. PMID- 12737798 TI - The role of laminin in embryonic cell polarization and tissue organization. AB - Genetic analyses have revealed that members of the laminin glycoprotein family are required for basement membrane assembly and cell polarization, with subsequent effects on cell survival and tissue organization during metazoan embryogenesis. These functions depend upon the cooperation between laminin polymerization and cell anchorage mediated via interactions with beta1-integrins, dystroglycan, and other cell surface receptors. PMID- 12737799 TI - The Caenorhabditis elegans hunchback-like gene lin-57/hbl-1 controls developmental time and is regulated by microRNAs. AB - Temporal control of development is an important aspect of pattern formation that awaits complete molecular analysis. We identified lin-57 as a member of the C. elegans heterochronic gene pathway, which ensures that postembryonic developmental events are appropriately timed. Loss of lin-57 function causes the hypodermis to terminally differentiate and acquire adult character prematurely. lin-57 is hbl-1, revealing a role for the worm hunchback homolog in control of developmental time. Significantly, fly hunchback (hb) temporally specifies cell fates in the nervous system. The hbl-1/lin-57 3'UTR is required for postembryonic downregulation in the hypodermis and nervous system and contains multiple putative binding sites for temporally regulated microRNAs, including let-7. Indeed, we find that hbl-1/lin-57 is regulated by let-7, at least in the nervous system. Examination of the hb 3'UTR reveals potential binding sites for known fly miRNAs. Thus, evolutionary conservation of hunchback genes may include temporal control of cell fate specification and microRNA-mediated regulation. PMID- 12737800 TI - The C elegans hunchback homolog, hbl-1, controls temporal patterning and is a probable microRNA target. AB - hunchback regulates the temporal identity of neuroblasts in Drosophila. Here we show that hbl-1, the C. elegans hunchback ortholog, also controls temporal patterning. Furthermore, hbl-1 is a probable target of microRNA regulation through its 3'UTR. hbl-1 loss-of-function causes the precocious expression of adult seam cell fates. This phenotype is similar to loss-of-function of lin-41, a known target of the let-7 microRNA. Like lin-41 mutations, hbl-1 loss-of-function partially suppresses a let-7 mutation. The hbl-1 3'UTR is both necessary and sufficient to downregulate a reporter gene during development, and the let-7 and lin-4 microRNAs are both required for HBL-1/GFP downregulation. Multiple elements in the hbl-1 3'UTR show complementarity to regulatory microRNAs, suggesting that microRNAs directly control hbl-1. MicroRNAs may likewise function to regulate Drosophila hunchback during temporal patterning of the nervous system. PMID- 12737801 TI - P58(IPK), a plant ortholog of double-stranded RNA-dependent protein kinase PKR inhibitor, functions in viral pathogenesis. AB - P58(IPK) is a cellular inhibitor of the mammalian double-stranded RNA-activated protein kinase (PKR). Here we provide evidence for the existence of its homolog in plants and its role in viral infection at the organism level. Viral infection of P58(IPK)-silenced Nicotiana benthamiana and Arabidopsis knockouts leads to host death. This host cell death is associated with phosphorylation of the alpha subunit of eukaryotic translation initiation factor (eIF-2alpha). Loss of P58(IPK) leads to reduced virus titer, suggesting that wild-type P58(IPK) protein plays an important role in viral pathogenesis. Although our complementation results using mammalian P58(IPK) suggest conservation of the P58(IPK) pathway in plants and animals, its biological significance seems to be different in these two systems. In animals, P58(IPK) is recruited by the influenza virus to limit PKR-mediated innate antiviral response. In plants, P58(IPK) is required by viruses for virulence and therefore functions as a susceptibility factor. PMID- 12737802 TI - FapR, a bacterial transcription factor involved in global regulation of membrane lipid biosynthesis. AB - Bacterial cells exert exquisite control over the biosynthesis of their membrane lipids, but the mechanisms are obscure. We describe the identification and purification from Bacillus subtilis of a transcription factor, FapR, that controls the expression of many genes involved in fatty acid and phospholipid metabolism (the fap regulon). Expression of this fap regulon is influenced by antibiotics that specifically inhibit the fatty acid biosynthetic pathway. We show that FapR negatively regulates fap expression and that the effects of antibiotics on fap expression are mediated by FapR. We further show that decreasing the cellular levels of malonyl-CoA, an essential molecule for fatty acid elongation, inhibits expression of the fap regulon and that this effect is FapR dependent. Our results indicate that control of FapR by the cellular pools of malonyl-CoA provides a mechanism for sensing the status of fatty acid biosynthesis and to adjust the expression of the fap regulon accordingly. PMID- 12737803 TI - Beta1 integrins regulate myoblast fusion and sarcomere assembly. AB - The mechanisms that regulate the formation of multinucleated muscle fibers from mononucleated myoblasts are not well understood. We show here that extracellular matrix (ECM) receptors of the beta1 integrin family regulate myoblast fusion. beta1-deficient myoblasts adhere to each other, but plasma membrane breakdown is defective. The integrin-associated tetraspanin CD9 that regulates cell fusion is no longer expressed at the cell surface of beta1-deficient myoblasts, suggesting that beta1 integrins regulate the formation of a protein complex important for fusion. Subsequent to fusion, beta1 integrins are required for the assembly of sarcomeres. Other ECM receptors such as the dystrophin glycoprotein complex are still expressed but cannot compensate for the loss of beta1 integrins, providing evidence that different ECM receptors have nonredundant functions in skeletal muscle fibers. PMID- 12737804 TI - Caspase activity and a specific cytochrome C are required for sperm differentiation in Drosophila. AB - The final stage of spermatid terminal differentiation involves the removal of their bulk cytoplasm in a process known as spermatid individualization. Here we show that apoptotic proteins play an essential role during spermatid individualization in Drosophila melanogaster. Several aspects of sperm terminal differentiation, including the activation of caspases, are reminiscent of apoptosis. Notably, caspase inhibitors prevent the removal of bulk cytoplasm in spermatids and block sperm maturation in vivo, causing male sterility. We further identified loss-of-function mutations in one of the two Drosophila cyt-c genes, cyt-c-d, which block caspase activation and subsequent spermatid terminal differentiation. Finally, a giant ubiquitin-conjugating enzyme, dBruce, is required to protect the sperm nucleus against hypercondensation and degeneration. These observations suggest that an apoptosis-like mechanism is required for spermatid differentiation in Drosophila. PMID- 12737805 TI - The COP9 signalosome promotes degradation of Cyclin E during early Drosophila oogenesis. AB - The COP9 signalosome (CSN) is an eight-subunit complex that regulates multiple signaling and cell cycle pathways. Here we link the CSN to the degradation of Cyclin E, which promotes the G1-S transition in the cell cycle and then is rapidly degraded by the ubiquitin-proteasome pathway. Using CSN4 and CSN5/Jab1 mutants, we show that the CSN acts during Drosophila oogenesis to remove Nedd8 from Cullin1, a subunit of the SCF ubiquitin ligase. Overexpression of Cyclin E causes similar defects as mutations in CSN or SCF(Ago) subunits: extra divisions or, in contrast, cell cycle arrest and polyploidy. Because the phenotypes are so similar and because CSN and Cyclin E mutations reciprocally suppress each other, Cyclin E appears to be the major target of the CSN during early oogenesis. Genetic interactions among CSN, SCF, and proteasome subunits further confirm CSN involvement in ubiquitin-mediated Cyclin E degradation. PMID- 12737806 TI - The Cdc14 phosphatase and the FEAR network control meiotic spindle disassembly and chromosome segregation. AB - During meiosis, DNA replication is followed by two consecutive rounds of chromosome segregation. Cells lacking the protein phosphatase CDC14 or its regulators, SPO12 and SLK19, undergo only a single meiotic division, with some chromosomes segregating reductionally and others equationally. We find that this abnormal chromosome behavior is due to an uncoupling of meiotic events. Anaphase I spindle disassembly is delayed in cdc14-1, slk19Delta, or spo12Delta mutants, but the chromosome segregation cycle continues, so that both meiotic chromosome segregation phases take place on the persisting meiosis I spindle. Our results show that Cdc14, Slk19, and Spo12 are not only required for meiosis I spindle disassembly but also play a pivotal role in establishing two consecutive chromosome segregation phases, a key feature of the meiotic cell cycle. PMID- 12737807 TI - Division of the nucleolus and its release of CDC14 during anaphase of meiosis I depends on separase, SPO12, and SLK19. AB - Disjunction of maternal and paternal centromeres during meiosis I requires crossing over between homologous chromatids, which creates chiasmata that hold homologs together. It also depends on a mechanism ensuring that maternal and paternal sister kinetochore pairs attach to oppositely oriented microtubules. Proteolytic cleavage of cohesin's Rec8 subunit by separase destroys cohesion between sister chromatid arms at anaphase I and thereby resolves chiasmata. The Spo12 and Slk19 proteins have been implicated in regulating meiosis I kinetochore orientation and/or in preventing cleavage of Rec8 at centromeres. We show here that the role of these proteins is instead to promote nucleolar segregation, including release of the Cdc14 phosphatase required for Cdk1 inactivation and disassembly of the anaphase I spindle. Separase is also required but surprisingly not its protease activity. It has two mechanistically different roles during meiosis I. Loss of the protease-independent function alone results in a second meiotic division occurring on anaphase I spindles in spo12delta and slk19delta mutants. PMID- 12737808 TI - Two redundant oscillatory mechanisms in the yeast cell cycle. AB - The cell division cycle requires oscillations in activity of B-type cyclin (Clb) Cdk1 kinases. Oscillations are due to periodic cyclin degradation by the anaphase promoting complex (APC) activated by Cdc20 or Cdh1, and to cyclical accumulation of the Sic1 inhibitor. The results presented here suggest that the regulatory machinery controlling Clb kinase levels embeds two distinct oscillatory mechanisms. One, a "relaxation oscillator," involves alternation between two meta stable states: Clb high/inhibitors (Sic1/APC-Cdh1) low, and Clb low/inhibitors high. The other, a "negative feedback oscillator," involves Clb kinase activation of APC-Cdc20, leading to Clb degradation. Genetic analysis suggests that these two mechanisms can function independently, and inactivation of both mechanisms is required to prevent mitosis. Computational modeling confirms that two such mechanisms can be linked to yield a robust cell cycle control system. PMID- 12737810 TI - The hexapeptide and linker regions of the AbdA Hox protein regulate its activating and repressive functions. AB - The Hox family transcription factors control diversified morphogenesis during development and evolution. They function in concert with Pbc cofactor proteins. Pbc proteins bind the Hox hexapeptide (HX) motif and are thereby thought to confer DNA binding specificity. Here we report that mutation of the AbdA HX motif does not alter its binding site selection but does modify its transregulatory properties in a gene-specific manner in vivo. We also show that a short, evolutionarily conserved motif, PFER, in the homeodomain-HX linker region acts together with the HX to control an AbdA activation/repression switch. Our in vivo data thus reveal functions not previously anticipated from in vitro analyses for the hexapeptide motif in the regulation of Hox activity. PMID- 12737809 TI - Syntaxin 2 and endobrevin are required for the terminal step of cytokinesis in mammalian cells. AB - The terminal step of cytokinesis in animal cells is the abscission of the midbody, a cytoplasmic bridge that connects the two prospective daughter cells. Here we show that two members of the SNARE membrane fusion machinery, syntaxin 2 and endobrevin/VAMP-8, specifically localize to the midbody during cytokinesis in mammalian cells. Inhibition of their function by overexpression of nonmembrane anchored mutants causes failure of cytokinesis leading to the formation of binucleated cells. Time-lapse microscopy shows that only midbody abscission but not further upstream events, such as furrowing, are affected. These results indicate that successful completion of cytokinesis requires a SNARE-mediated membrane fusion event and that this requirement is distinct from exocytic events that may be involved in prior ingression of the plasma membrane. PMID- 12737811 TI - Splicing factor 1 in the pocket. PMID- 12737812 TI - Examining both sides of a Janus PTB domain. PMID- 12737813 TI - Pushing induced fit to its limits: tRNA-dependent active site assembly in class I aminoacyl-tRNA synthetases. PMID- 12737814 TI - Folding by consensus. PMID- 12737815 TI - Structural insights into the origins of DNA polymerase fidelity. AB - DNA polymerases discriminate from a pool of structurally similar molecules to insert the correct nucleotide to preserve Watson-Crick base pairing rules. The ability to choose between "right and wrong" is highly dependent on the identity of the polymerase. Because naturally occurring polymerases with divergent fidelities insert incorrect nucleotides with comparable efficiencies, fidelity is primarily governed by the ability to insert the correct nucleotide. DNA polymerases generally bind the correct nucleotide with similar affinities, but low-fidelity polymerases insert correct nucleotides more slowly than higher fidelity enzymes. A comparison of crystallographic ternary substrate complexes of DNA polymerases from five families exhibiting a range of nucleotide insertion rates reveals possible structural features that lead to rapid, efficient, and faithful DNA synthesis. PMID- 12737816 TI - Design of stable alpha-helical arrays from an idealized TPR motif. AB - The tetratricopeptide repeat (TPR) is a 34-amino acid alpha-helical motif that occurs in over 300 different proteins. In the different proteins, three to sixteen or more TPR motifs occur in tandem arrays and function to mediate protein protein interactions. The binding specificity of each TPR protein is different, although the underlying structural motif is the same. Here we describe a statistical approach to the design of an idealized TPR motif. We present the high resolution X-ray crystal structures (to 1.55 and 1.6 A) of designed TPR proteins and describe their solution properties and stability. A detailed analysis of these structures provides an understanding of the TPR motif, how it is repeated to give helical arrays with different superhelical twists, and how a very stable framework may be constructed for future functional designs. PMID- 12737817 TI - Structure of the predominant protein arginine methyltransferase PRMT1 and analysis of its binding to substrate peptides. AB - PRMT1 is the predominant type I protein arginine methyltransferase in mammals and highly conserved among all eukaryotes. It is essential for early postimplantation development in mouse. Here we describe the crystal structure of rat PRMT1 in complex with the reaction product AdoHcy and a 19 residue substrate peptide containing three arginines. The results reveal a two-domain structure-an AdoMet binding domain and a barrel-like domain-with the active site pocket located between the two domains. Mutagenesis studies confirmed that two active site glutamates are essential for enzymatic activity, and that dimerization of PRMT1 is essential for AdoMet binding. Three peptide binding channels are identified: two are between the two domains, and the third is on the surface perpendicular to the strands forming the beta barrel. PMID- 12737818 TI - Crystal structures of oligomeric forms of the IP-10/CXCL10 chemokine. AB - We have determined the structure of wild-type IP-10 from three crystal forms. The crystals provide eight separate models of the IP-10 chain, all differing substantially from a monomeric IP-10 variant examined previously by NMR spectroscopy. In each crystal form, IP-10 chains form conventional beta sheet dimers, which, in turn, form a distinct tetrameric assembly. The M form tetramer is reminiscent of platelet factor 4, whereas the T and H forms feature a novel twelve-stranded beta sheet. Analytical ultracentrifugation indicates that, in free solution, IP-10 exists in a monomer-dimer equilibrium with a dissociation constant of 9 microM. We propose that the tetrameric structures may represent species promoted by the binding of glycosaminoglycans. The binding sites for several IP-10-neutralizing mAbs have also been mapped. PMID- 12737819 TI - Structural basis of the highly efficient trapping of the HIV Tat protein by an RNA aptamer. AB - An RNA aptamer containing two binding sites exhibits extremely high affinity to the HIV Tat protein. We have determined the structure of the aptamer complexed with two argininamide molecules. Two adjacent U:A:U base triples were formed, which widens the major groove to make space for the two argininamide molecules. The argininamide molecules bind to the G bases through hydrogen bonds. The binding is stabilized through stacking interactions. The structure of the aptamer complexed with a Tat-derived arginine-rich peptide was also characterized. It was suggested that the aptamer structure is similar for both complexes and that the aptamer interacts with two different arginine residues of the peptide simultaneously at the two binding sites, which could explain the high affinity to Tat. PMID- 12737820 TI - The structure of Acidithiobacillus ferrooxidans c(4)-cytochrome: a model for complex-induced electron transfer tuning. AB - The study of electron transfer between the copper protein rusticyanin (RCy) and the c(4)-cytochrome CYC(41) of the acidophilic bacterium Acidithiobacillus ferrooxidans has evidenced a remarkable decrease of RCy's redox potential upon complex formation. The structure of the CYC(41) obtained at 2.2 A resolution highlighted a specific glutamate residue (E121) involved in zinc binding as potentially playing a central role in this effect, required for the electron transfer to occur. EPR and stopped-flow experiments confirmed the strong inhibitory effect of divalent cations on CYC(41):RCy complex formation. A docking analysis of the CYC(41) and RCy structure allows us to propose a detailed model for the complex-induced tuning of electron transfer in agreement with our experimental data, which could be representative of other copper proteins involved in electron transfer. PMID- 12737821 TI - Structure-guided design of sialic acid-based Siglec inhibitors and crystallographic analysis in complex with sialoadhesin. AB - The Siglec family of receptors mediates cell surface interactions through recognition of sialylated glycoconjugates. The crystal structure of the N terminal immunoglobulin-like domain of the Siglec sialoadhesin (SnD1) in complex with 2,3-sialyllactose has informed the design of sialic acid analogs (sialosides) that bind Siglecs with significantly enhanced affinities and specificities. Binding assays against sialoadhesin (Sn; Siglec-1), CD22 (Siglec 2), and MAG (Siglec-4) show a 10- to 300-fold reduction in IC(50) values (relative to methyl-alpha-Neu5Ac) for three sialosides bearing aromatic group modifications of the glycerol side chain: Me-alpha-9-N-benzoyl-amino-9-deoxy Neu5Ac (BENZ), Me-alpha-9-N-(naphthyl-2-carbonyl)-amino-9-deoxy-Neu5Ac (NAP), and Me-alpha-9-N-(biphenyl-4-carbonyl)-amino-9-deoxy-Neu5Ac (BIP). Crystal structures of these sialosides in complex with SnD1 suggest explanations for the differences in specificity and affinity, providing further ideas for compound design of physiological and potentially therapeutic relevance. PMID- 12737822 TI - Origins of peptide selectivity and phosphoinositide binding revealed by structures of disabled-1 PTB domain complexes. AB - Formation of the mammalian six-layered neocortex depends on a signaling pathway that involves Reelin, the very low-density lipoprotein receptor, the apolipoprotein E receptor-2 (ApoER2), and the adaptor protein Disabled-1 (Dab1). The 1.5 A crystal structure of a complex between the Dab1 phosphotyrosine binding (PTB) domain and a 14-residue peptide from the ApoER2 tail explains the unusual preference of Dab1 for unphosphorylated tyrosine within the NPxY motif of the peptide. Crystals of the complex soaked with the phosphoinositide PI-4,5P(2) (PI) show that PI binds to conserved basic residues on the PTB domain opposite the peptide binding groove. This finding explains how the Dab1 PTB domain can simultaneously bind PI and the ApoER2 tail. Recruitment of the Dab1 PTB domain to PI-rich regions of the plasma membrane may facilitate association with the Reelin receptor cytoplasmic tails to transduce a critical positional cue to migrating neurons. PMID- 12737823 TI - De novo design of foldable proteins with smooth folding funnel: automated negative design and experimental verification. AB - De novo sequence design of foldable proteins provides a way of investigating principles of protein architecture. We performed fully automated sequence design for a target structure having a three-helix bundle topology and synthesized the designed sequences. Our design principle is different from the conventional approach, in that instead of optimizing interactions within the target structure, we design the global shape of the protein folding funnel. This includes automated implementation of negative design by explicitly requiring higher free energy of the denatured state. The designed sequences do not have significant similarity to those of any natural proteins. The NMR and CD spectroscopic data indicated that one designed sequence has a well-defined three-dimensional structure as well as alpha-helical content consistent with the target. PMID- 12737824 TI - tRNA-dependent active site assembly in a class I aminoacyl-tRNA synthetase. AB - The crystal structure of ligand-free E. coli glutaminyl-tRNA synthetase (GlnRS) at 2.4 A resolution shows that substrate binding is essential to construction of a catalytically proficient active site. tRNA binding generates structural changes throughout the enzyme, repositioning key active site peptides that bind glutamine and ATP. The structure gives insight into longstanding questions regarding the tRNA dependence of glutaminyl adenylate formation, the coupling of amino acid and tRNA selectivities, and the roles of specific pathways for transmission of tRNA binding signals to the active site. Comparative analysis of the unliganded and tRNA-bound structures shows, in detail, how flexibility is built into the enzyme architecture and suggests that the induced-fit transitions are a key underlying determinant of both amino acid and tRNA specificity. PMID- 12737825 TI - Epilepsy and the Internet. PMID- 12737826 TI - An Approach to Classifying Neuropsychiatric Disorders in Epilepsy. PMID- 12737828 TI - The Frontal Lobes, Epilepsy, and Behavior. AB - The frontal lobes have been overshadowed by the temporal lobes in the vast literature addressing the neurobehavioral and psychological perspectives of epilepsy. The purpose of this review is to summarize contemporary anatomicobehavioral correlations and to highlight the frontal lobe contributions to the neurology, neuropsychology, and neuropsychiatry of epilepsy, in general, and to temporal lobe epilepsy (TLE) and frontal lobe epilepsy (FLE), in particular. Much evidence has accumulated suggesting that focal epileptogenic tissue may have effects on distant neural systems. Data supporting the case that the frontal regions are preferentially affected in TLE are presented. Emphasis is placed on the results of numerous functional imaging studies demonstrating correlations between frontal hypoperfusion and cognitive or mood impairments in patients with TLE. PMID- 12737827 TI - Effects of Anticonvulsant Medication on EEG Sleep Architecture. PMID- 12737829 TI - Oxcarbazepine for Treatment of Partial Epilepsy: A Review and Recommendations for Clinical Use. AB - Recent trials and extensive postmarketing use confirm the efficacy and safety of oxcarbazepine (OXC) as a first-line treatment for adults and children with simple partial seizures, complex partial seizures, and partial seizures evolving to secondarily generalized seizures. OXC undergoes reductive metabolism at its keto moiety to form 10-hydroxy-10,11-dihydro-10-oxo-5H-dibenz[b,f]azepine-5 carboxamide (MHD), which is glucuronidated and excreted in the urine, with minimal involvement of the hepatic cytochrome P450-dependent enzymes. OXC has some drug interactions, and does not require titration, allowing for better tolerability. Titration for monotherapy and adjunctive therapy of OXC could begin at 150 mg/day and be increased by 150 mg every 2-3 days until the target dose of 900-1200 mg/day is reached. If necessary, one can go faster and start with up to 600 mg/day and titrate with weekly increments up to 600 mg/day if necessary for seizure control. Conversion to monotherapy can be done overnight or gradually. For gradual conversion, use the recommended titration of OXC and withdraw the baseline antiepileptic drugs gradually by 25%, starting at Day 14 or earlier in case of baseline tolerability issues. Consider reducing the dose of the primary antiepileptic drug during adjunctive therapy in case of adverse events or increase the dose of OXC in case of incomplete seizure control. In children OXC should be started at 8-10 mg/kg/day in two or three divided doses. If clinically indicated the dose can then be increased by 10 mg/kg/day in weekly intervals with final doses up to 30-46 mg/kg/day. Dose adjustment may be necessary in very young children (age 2-5 years) and in patients with renal dysfunction, based on renal clearance. However no adjustment of OXC dose is needed in patients with mild to moderate hepatic dysfunction. OXC has a number of advantages which include rapid titration, no need for safety monitoring (except for uncommon and mostly asymptomatic hyponatremia), a low potential for drug-drug interactions (except for those possibly impairing the effectiveness of oral contraceptives and increasing the serum concentration of phenytoin), a rash rate of less than 5%, similar efficacy and similar or better tolerability and safety compared with first-generation antiepileptic drugs. OXC is a valuable alternative to current treatment options. PMID- 12737830 TI - Adolescent and Maternal Perspectives of Quality of Life and Neuropsychological Status Following Epilepsy Surgery. AB - A convenience sample of 13 adolescents who had undergone epilepsy surgery and their mothers reported their perceptions of change in areas affecting quality of life (physical, affect/behavior, social, cognition/academics, and family). A mixed method approach was used, with self-report and maternal-report narratives (qualitative measures), as well as neuropsychological profiles (quantitative measures). The results demonstrate that change after epilepsy surgery is multifaceted, and that adolescent and maternal perceptions are notably different. Whereas most adolescents experienced positive changes, mothers simultaneously reported many more negative changes along with positive changes. Furthermore, mothers frequently described aspects of their adolescent's life that continued to be of concern, whereas adolescents did not. The findings strongly suggest that the processes involved in postsurgical adjustment are complex. The development of a theoretical model is essential to help build an understanding of factors that contribute to positive and negative outcomes. PMID- 12737831 TI - Nonconvulsive Kainic Acid-Induced Seizures Elicit Age-Dependent Impairment of Memory for the Elevated Plus-Maze. AB - The goal of this study was to evaluate changes in spatial learning in adult and immature rats during and after nonconvulsive seizures. An elevated plus-maze was used in 18- and 25-day-old and adult rats. Kainic acid (KA 6 mg/kg) was administered 60 minutes before the first exposure (Experiment 1) or after a 3-day pretraining (Experiment 2, only adult rats). Animals were retested three times with 24-hour intervals. EEG activity was monitored in 18-day-old rats. KA prolonged the transfer latency (TL) in all age groups. In the youngest group the TL was prolonged 24 hours after KA when epileptic EEG graphoelements were still registered. In both older groups, prolonged TL was measured only 60 minutes after KA. In the pretrained adults, significantly prolonged TLs persisted for 24 hours after KA. KA changed the performance of adult and immature rats in the elevated plus maze not only during nonconvulsive seizures but also 24 hours later. PMID- 12737832 TI - Informant Agreement in Behavior Ratings for Children with Epilepsy. AB - The purpose of this study was to determine the degree of informant agreement for behavioral ratings of children with epilepsy. Informants completed Achenbach's 1991 scales: parents completed the Child Behavior Checklist (CBCL), teachers completed the Teacher's Report Form (TRF), and youth completed the Youth Self Report Form of the CBCL (YSR). Analyses included degree of concordance of ratings as a function of informant, child gender, and condition severity (active vs inactive epilepsy). Results indicated that across all four types of raters (mothers, fathers, teachers, and adolescents) there was a similar pattern. Mothers' ratings tended to be the highest and youths' ratings tended to be the lowest across scales. In general, agreement among adult raters was greater than between youth and adults. Mothers and teachers reported more internalizing symptoms than did youth; mother, father, and teacher ratings on externalizing symptoms were not significantly different from each other. There were no significant effects of gender and condition severity on concordance among ratings although there were some interesting trends. PMID- 12737833 TI - De Novo Nonepileptic Seizures after Cranial Surgery for Epilepsy: Incidence and Risk Factors. AB - We evaluated the incidence of de novo nonepileptic seizures (NES), confirmed by EEG monitoring, after cranial surgery for intractable epilepsy in 228 surgery patients. Eight patients (3.5%) developed de novo NES at 6 weeks to 6 years (mean, 23 months) after surgery. Six had undergone a resection and two complete callosotomy. They did not differ from a larger surgical group with respect to sex, side of surgery, age at onset, or duration of epilepsy, Full Scale Intelligence Quotient, seizure outcome, or preoperative interictal dysphoric disorder (IDD), but there was a significant excess of postoperative IDD and operative complications (bone flap infections); the callosotomy patients had marked hemisphere disconnection syndromes. Repeat EEG videotelemetry monitoring is important to detect postoperative NES so that inappropriate therapeutic measures may be avoided. Risk factors may be exacerbation or persistence of IDD and surgical complications. The etiology of NES is discussed. PMID- 12737834 TI - Treatment of Interictal Depression with Citalopram in Patients with Epilepsy. AB - The purpose of this study is to assess the efficacy and safety of the selective serotonin-reuptake inhibitor (SSRI) citalopram in depressed epileptic patients. We evaluated 43 epileptic patients who suffered from depression and whose total score on the 21 items of the Hamilton Scale for Depression (HAMD 21) exceeded 15 points. These patients were examined by the psychiatrist and scaled before treatment and after 4 and 8 weeks of treatment with citalopram. The dose of citalopram was flexible, related to the actual condition of the patient. In each patient and in the whole group of patients we compared the monthly seizure frequency (total, partial seizures, generalized tonic-clonic seizures) recorded during treatment with citalopram with that recorded during the 2 months preceding the start of citalopram. During treatment we observed a decrease in the total score on the HAMD 21 from a mean initial value of 21.5 +/- 2.9 (range, 17-26) prior to therapy 14.5 +/- 2.9 (range, 10-19) (P < 0.001) after 4 weeks of treatment and to 9.9 +/- 3.1 (range, 4-19) (P < 0.001) after 8 weeks of treatment. There were 9 (20.9%) responders after 4 weeks of treatment and 28 responders (65.1%) after 8 weeks, all of them with decrease on the HAMD 21 greater than 50%. Nausea was the most common adverse event in 7 patients (16.3%) during the first month of treatment and in 3 patients (6.9%) during the second month of treatment. Sexual dysfunction (decrease of libido) was reported in 2 (4.7%) male patients during the entire course of treatment. No seizure worsening was observed in our patients. Monthly seizure frequency did not change significantly: 2.24 (+/-0.76) seizures before treatment with citalopram, 2.29 (+/ 0.81) seizures in the first month of treatment, 2.21 (+/-1.00) seizures in the second month of treatment. No occurrence of de novo generalized tonic-clonic seizures was recorded in individual patients. Citalopram is a safe and effective antidepressant in the treatment of depressed epileptic patients. PMID- 12737835 TI - Fenfluramine-Phentermine (Fen-Phen) and Seizures: Evidence for an Association. AB - Fenfluramine-phentermine combination therapy ("fen-phen") became a popular treatment for obesity in the 1990s. Although this treatment causes cardiac toxicity, use of these medications has not previously been associated with seizures. We report five cases with apparent association between use of fenfluramine-phentermine and occurrence of seizures. Three patients with a history of childhood-onset idiopathic generalized epilepsy in remission experienced a recrudescence of seizures following treatment with fenfluramine phentermine. Two patients presented with new-onset seizures in midlife following use of fenfluramine-phentermine, and seizures persisted following discontinuation of this therapy. One of these patients restarted fenfluramine-phentermine months later, and experienced recurrent seizures. The nature of the association between fenfluramine-phentermine and seizures is uncertain from this preliminary report. There may be a specific association with idiopathic generalized epilepsies, which appeared to be overrepresented in this case series. An effect of fen-phen on seizure threshold appears most likely; however, an epileptogenic effect cannot be excluded. PMID- 12737836 TI - Attention Deficit-Hyperactivity Disorder and the Electroencephalogram. To the Editor. PMID- 12737837 TI - Lying: A Metaphorical Memoir. By Lauren Slater. PMID- 12737838 TI - Oxcarbazepine in Treatment of Painful Peripheral Neuropathies. PMID- 12737839 TI - Design and synthesis of pH-responsive polymeric carriers that target uptake and enhance the intracellular delivery of oligonucleotides. AB - The delivery of biomolecular therapeutics that function intracellularly remains a significant challenge in the field of biotechnology. In this report, a new family of polymeric drug carriers that combine cell targeting, a pH-responsive membrane disruptive component, and serum-stabilizing polyethylene glycol (PEG) grafts, is shown to direct the uptake and endosomal release of oligonucleotides in a primary hepatocyte cell line. These polymers are called encrypted polymers and are graft terpolymers that consist of a hydrophobic, membrane-disruptive backbone onto which hydrophilic PEG chains have been grafted through acid-degradable linker acetal linkages. In this report, the ability of the encrypted polymers to deliver rhodamine-labeled oligonucleotides or PEG-FITC (a model macromolecular drug) (5 kDa) into the cytoplasm of hepatocytes was investigated by fluorescence microscopy. Two new encrypted polymer derivatives (polymers E2 and E3) were synthesized that contained lactose for targeting to hepatocytes. Polymer E2 also has PEG-FITC conjugated to it, as a model macromolecular drug, and polymer E3 contains a pendant hexalysine moiety for complexing oligonucleotides. The results of the fluorescence microscopy experiments show that the encrypted polymers direct vesicular escape and efficiently deliver oligonucleotides and macromolecules into the cytoplasm of hepatocytes. PMID- 12737840 TI - Correlation of characteristics of gel extrusion module (GEM) tablet formulation and drug dissolution rate. AB - The purpose of the study was to characterize the swelling properties of controlled release tablet formulations with compositional and processing differences. A correlation was also established between the drug dissolution rate from the controlled release gel extrusion module (GEM) tablet and the swelling properties of the core tablet. The GEM tablet consisted of a core tablet of water swellable Carbopol polymer, a neutralizing agent, drug, and excipients. The tablet was subsequently coated with a rigid, water impermeable membrane. A number of holes were then drilled by laser through the impermeable membrane. Dynamic mechanical analysis (DMA), Perkin Elmer DMA7, was used to characterize the swelling properties. During the swelling measurements, the measuring probe and sample were completely submerged in the surrounding medium. The results showed that the formulation containing potassium phosphate dibasic as a neutralizing agent had the highest swelling rate. Correspondingly, this formulation had the highest drug dissolution rate over the same time period. The processing difference included wet granulation and dry roller compaction. The compositional differences included different neutralizing agents, binders or Carbopol polymers. The results showed a linear relationship (r(2)=0.954) between the swelling rates of the core tablets and the drug dissolution rates of GEM tablets. No correlation was found between the drug dissolution rates and either the maximum extent of tablet swelling or the time needed to reach the maximum extent of swelling. The results demonstrated that DMA could be used to support both formulation and process development to determine the effects of different compositions and manufacturing processes on drug dissolution rates for swelling controlled release devices. PMID- 12737841 TI - Press-coating of immediate release powders onto coated controlled release tablets with adhesives. AB - A novel adhesive coating was developed that allows even small quantities of immediate-release (IR) powders to be press-coated onto controlled-release (CR), coated dosage forms without damaging the CR coating. The process was exemplified using a pseudoephedrine osmotic tablet (asymmetric membrane technology, AMT) where a powder weighing less than 25% of the core was pressed onto the osmotic tablet providing a final combination tablet with low friability. The dosage form with the adhesive plus the press-coated powder showed comparable sustained drug release rates to the untreated dosage form after an initial 2-h lag. The adhesive layer consisted of an approximately 100- microm coating of Eudragit RL, polyethylene glycol (PEG) and triethyl citrate (TEC) at a ratio of 5:3:1.2. This coating provides a practical balance between handleability before press-coating and good adhesion. PMID- 12737842 TI - Cisplatin incorporated in microspheres: development and fundamental studies for its clinical application. AB - A new drug delivery formulation, biodegradable glycolic acid-lactic acid copolymer (PGLA) microspheres incorporating cisplatin (CDDP-MS) has been developed for the treatment of peritoneal carcinomatosis. Scanning electron microscopy showed that CDDP-MS has a smooth surface and few cisplatin crystals in the hollow. An electron probe micro analyzer revealed that cisplatin was located mainly in the matrix in the state of a molecule. Release profile in vitro of CDDP from microspheres showed that the initial burst was 21.2% and the remaining CDDP was released slowly thenceforth over 14 days. Hydrolysis of CDDP-MS progresses very slowly during the 14 days, but there was no morphological change in the SEM views. The dimethylformamide content entrapped within CDDP-MS, determined by a gas chromatography, was 136 ppm at the evaporation temperature of 47 degrees C. The 50% lethal dose value of CDDP-MS, calculated by the Litchfield-Wilcoxon method, was reduced to 57% of the cisplatin solution. Therapeutic experiment on mice with peritoneal carcinomatosis showed that CDDP-MS did not enhance therapeutic effect as compared with the same dose dosage of a cisplatin aqueous solution but large quantities of cisplatin could be given in case of CDDP-MS owing to less toxicity. PMID- 12737843 TI - Transscleral permeability and intraocular concentrations of cisplatin from a collagen matrix. AB - This study determined the in vitro permeability of cisplatin through isolated human sclera as delivered by a collagen matrix vehicle. Short-term and long-term intraocular levels of cisplatin were also measured in the rabbit eye after a subconjunctival injection. Cisplatin in either a collagen matrix vehicle or a control balanced salt solution (BSS) vehicle was applied to human sclera mounted in a specially designed in vitro perfusion chamber. The amount of cisplatin that diffused across the sclera was measured in hourly samples for 24 h using atomic absorption spectrometry. In vivo studies were also performed in Dutch Belted rabbits given subconjunctival injections of cisplatin in collagen matrix or in BSS. Eyes were enucleated at 1.5 h and 2 weeks after injection, frozen, and dissected to determine the intraocular cisplatin concentrations. Cisplatin had a peak in vitro scleral permeability constant of 8.3+/-1.2 x 10(-6) and 20.1+/-1.8 x 10(-6) cm/s, delivered in collagen matrix and in BSS, respectively (mean+/ S.D.). At the end of the in vitro experiments, 35.9+/-4.6% of the cisplatin remained in the collagen matrix, while 0.8+/-0.2% remained in the BSS vehicle. Subconjunctival injection of cisplatin in the collagen matrix vehicle achieved 3.3+/-0.1 microg/ml in the vitreous humor at 1.5 h and 0.1+/-0.1 microg/ml at 2 weeks. This vehicle also achieved a cisplatin concentration of 73.5+/-23.9 microg/mg in the choroid and retina at 1.5 h and 3.2+/-1.3 microg/mg at 2 weeks. Compared to BSS, the collagen matrix vehicle provided a more controlled release of cisplatin, and after subconjunctival injection into rabbits, attained higher drug levels in several ocular tissues. PMID- 12737844 TI - Mucoadhesive ocular insert based on thiolated poly(acrylic acid): development and in vivo evaluation in humans. AB - The aim of the study was to develop a mucoadhesive ocular insert for the controlled delivery of ophthalmic drugs and to evaluate its efficacy in vivo. The inserts tested were based either on unmodified or thiolated poly(acrylic acid). Water uptake and swelling behavior of the inserts as well as the drug release rates of the model drugs fluorescein and two diclofenac salts with different solubility properties were evaluated in vitro. Fluorescein was used as fluorescent tracer to study the drug release from the insert in humans. The mean fluorescein concentration in the cornea/tearfilm compartment as a function of time was determined after application of aqueous eye drops and inserts composed of unmodified and of thiolated poly(acrylic acid). The acceptability of the inserts by the volunteers was also evaluated. Inserts based on thiolated poly(acrylic acid) were not soluble and had good cohesive properties. A controlled release was achieved for the incorporated model drugs. The in vivo study showed that inserts based on thiolated poly(acrylic acid) provide a fluorescein concentration on the eye surface for more than 8 h, whereas the fluorescein concentration rapidly decreased after application of aqueous eye drops or inserts based on unmodified poly(acrylic acid). Moreover, these inserts were well accepted by the volunteers. The present study indicates that ocular inserts based on thiolated poly(acrylic acid) are promising new solid devices for ocular drug delivery. PMID- 12737845 TI - Potentiality of double liposomes containing salmon calcitonin as an oral dosage form. AB - The hypocalcemic effects of salmon calcitonin (SCT) after oral administration in rats by means of SCT-loading double liposomes (DL) which consist of liposomes containing small liposomes were investigated. SCT-loading DL consisted of four types of the inner liposomes such as neutral liposomes (NL) and cationic charged liposomes (CL) prepared using Coatsome, and neutral (VET) and cationic charged (c VET) liposomes prepared using a mechanochemical method and sizing to 100 nm by the extrusion procedure were prepared. DL could be prepared by a combination of mechanochemical and glass-beads methods at a high efficiency. DL produced the increase in bioavailability in all groups treated with SCT-loading liposomes except for c-VET. The bioavailability of VET-DL was not significantly different but the greatest among the samples used in this study regardless of the similar size of NL-DL and CL-DL, and was approx. 6.8-fold higher than that of SCT solution when taken orally. The group treated with c-VET showed the strongest hypocalcemic effects among the inner liposomes examined (P>0.05). Therefore, it is speculated that not only the size of liposomes but also the cationic charge plays an important role in the intestinal absorption of DL. These findings suggested the utility of DL as an oral dosage form of SCT. PMID- 12737846 TI - Polymeric micelles of poly(2-ethyl-2-oxazoline)-block-poly(epsilon-caprolactone) copolymer as a carrier for paclitaxel. AB - Polymeric micelles based on amphiphilic block copolymers of poly(2-ethyl-2 oxazoline) (PEtOz) and poly(epsilon -caprolactone) (PCL) were prepared in an aqueous phase. The loading of paclitaxel into PEtOz-PCL micelles was confirmed by 1H-NMR spectra. Paclitaxel was efficiently loaded into PEtOz-PCL micelles using dialysis method, and the loading content of paclitaxel in micelles was in the range 0.5-7.6 wt.% depending on the block composition of block copolymers, organic solvent used in the dialysis, and feed weight ratio of paclitaxel to block copolymer. The higher the content of hydrophobic block in the block copolymers, the higher the loading efficiency of micelles for paclitaxel. When acetonitrile was used as solvent, a higher drug loading efficiency was obtained than with THF. The loading efficiency decreased with increasing feed weight ratio of paclitaxel to block copolymer from 0.1:1 to 0.2:1. The hydrodynamic diameters of paclitaxel-loaded micelles were in the range 18.3-23.4 nm with narrow size distribution. The hemolysis test of PEtOz-PCL performed in vitro indicated that the toxicity of PEtOz-PCLs to lipid membrane was not significant compared with Tween 80, and was comparable to that observed with Cremophore EL. The proliferation inhibition activity of paclitaxel-loaded micelles for KB human epidermoid carcinoma cells was also evaluated in vitro. Paclitaxel-entrapped polymeric micelles exhibited comparable activity to that observed with Cremophore EL-based paclitaxel formulations in inhibiting the growth of KB cells. PMID- 12737847 TI - Effect of gamma-irradiation on cladribine and cladribine-containing biodegradable copolymers. AB - The aims of this study were to assess the effects of sterilization with gamma irradiation on (i). bulk cladribine and (ii). cladribine-containing biodegradable copolymers. The stability of cladribine upon irradiation was confirmed by TLC, HPLC, UV, IR, DSC, rentgenography and electron microscopy. The stability of copolymers containing cladribine upon irradiation was assessed by IR, DSC and EPR. In vitro kinetics of nucleoside release from the copolymers before and after irradiation were compared, and only slight changes were found. Results of our study indicate that gamma-irradiation can be safely applied for the sterilization of cladribine or cladribine-containing copolymers for medical purposes. PMID- 12737848 TI - Blends of enteric and GIT-insoluble polymers used for film coating: physicochemical characterization and drug release patterns. AB - THE OBJECTIVES OF THIS STUDY WERE: (i). to use blends of gastrointestinal tract (GIT)-insoluble and enteric polymers (ethyl cellulose and Eudragit L) as coating materials for multiparticulate controlled release dosage forms; (ii). to investigate the effects of the polymer blend ratio and coating level on the resulting drug release patterns; and (iii). to explain the observed phenomena based on the physicochemical properties of the systems. Propranolol HCl-loaded pellets were coated in a fluidized bed coater with organic polymer solutions; thin, drug-containing and drug-free, polymeric films were prepared using a casting knife. In vitro drug release, water uptake and dry weight loss studies were performed in 0.1 M HCl and phosphate buffer pH 7.4, respectively. The apparent drug diffusion coefficients within the polymeric systems were determined using different experimental and theoretical techniques (side-by-side diffusion cells, in vitro drug release from thin films; exact and approximate solutions of Fick's second law of diffusion). A broad range of drug release patterns from coated pellets could be achieved by varying the GIT-insoluble:enteric polymer blend ratio. With increasing relative amounts of Eudragit L, the release rates in both media significantly increased. The increase at low pH could be attributed to an increase in water uptake, as observed with thin films. Interestingly, only partial Eudragit L leaching occurred in phosphate buffer pH 7.4 even at high enteric polymer contents, indicating that the GIT-insoluble polymer effectively hindered the dissolution of the entrapped Eudragit L. At high pH, both polymer leaching and polymer swelling contributed to the control of drug release. The determined apparent drug diffusion coefficients take the two effects adequately into account. PMID- 12737849 TI - Cationic emulsions improves the delivery of oligonucleotides to leukemic P388/ADR cells in ascite. AB - The aim of this study was to investigate the in vivo ability of O/W cationic emulsions to deliver oligonucleotides (ON) in leukemic P388/ADR cells in ascite, after intraperitoneal (IP) administration in mice. Cationic emulsions were prepared by microfluidization as previously described by Teixeira et al. [Pharm. Res 16 (1999) 30]. The formulations consisted mainly of medium chain triglycerides, phosphatidylcholine (PC), poloxamer, and either a monocationic lipid stearylamine (PC/SA-emulsion) or a polycationic lipid RPRC(18) (PC/RPRC(18) emulsion). A model ON (33P-pdT(16)) was associated with cationic emulsions by single addition at the end of the manufacturing process. Seven days after P388/ADR inoculation IP to mice, ON free or associated with PC/SA or PC/RPRC(18) emulsions was injected IP at a dose of 0.5 mg/kg. At different interval times, ascite including cells, blood and the main organs were collected and the radioactivity counted by liquid scintillation. The overall results showed significantly high amounts of ON in the leukemic cell pellet, 24 h after administration of ON associated to either PC/SA (AUC(0-24 h)=13634, %injected dose/min) or PC/RPRC(18) (AUC(0-24 h)=22592, % injected dose/min), contrary to the free ON solution (AUC(0-24 h)=3095, %injected dose/min), which displayed only reduced capture by cancer cells. In conclusion, complexation of ON with cationic emulsions had a beneficial effect in increasing tumor cells uptake in vivo (up to sevenfold for PC/RPRC(18)-emulsion) after IP administration. This could open interesting prospects for the treatment of ovarian cancers. PMID- 12737850 TI - Water-soluble biodegradable cationic polyphosphazenes for gene delivery. AB - Polyphosphazenes bearing cationic moieties were synthesized from poly(dichloro)phosphazene, which in turn was obtained by thermal polymerization of hexachlorocyclotriphosphazene in 1,2,4-trichlorobenzene. Next, either 2 dimethylaminoethanol (DMAE) or 2-dimethylaminoethylamine (DMAEA) side groups were introduced by a substitution reaction. The polymers were purified by dialysis against water and tetrahydrofuran, lyophilized and evaluated as polymeric transfectants. The polyphosphazenes were able to bind plasmid DNA yielding positively charged particles (polyplexes) with a size around 80 nm at a polymer/DNA ratio of 3:1 (w/w). The polyphosphazene-based polyplexes were able to transfect COS-7 cells in vitro with an efficiency comparable to a well-known polymeric transfectant [poly(2-dimethylaminoethyl methacrylate), pDMAEMA]. The toxicity of both polyphosphazenes was lower than pDMAEMA. The transfection efficiency for the poly(DMAE)phosphazene-based polyplexes was about threefold higher in the absence of serum than in the presence of 5.0% fetal bovine serum. This is probably caused by unfavorable interactions of the polyplexes with serum proteins. In contrast, the poly(DMAEA)phosphazene-based polyplexes showed a threefold lower transfection activity in the absence of serum. For this system, serum proteins likely masked the toxicity of the polyplexes, as shown by the XTT cell viability assay and confocal laser scanning microscopy studies. Preliminary degradation studies indicate that the polymers were indeed degradable. The half life at pH 7.5 and 37 degrees C was around 7 days for poly(DMAE)phosphazenes and 24 days for poly(DMAEA)phosphazenes. This study shows that polyphosphazenes are a suitable and promising new class of biodegradable polymeric carriers for gene delivery. PMID- 12737852 TI - Will SARS hurt the world's poor? PMID- 12737851 TI - Humanized docking system for assembly of targeting drug delivery complexes. AB - Targeted drug delivery requires 'loading' drugs onto targeting proteins. Traditional technologies for loading drugs rely on chemical conjugation of drugs or drug carriers to targeting proteins. An alternative approach might rely on assembly of targeting complexes using a docking system that includes two components: a 'docking' tag fused to a targeting protein, and a 'payload' module containing an adapter protein for non-covalent binding to the docking tag. We describe here a fully humanized adapter/docking tag system based on non-covalent interaction between two fragments of human pancreatic RNase I. A 15 amino acid long N-terminal fragment of RNase I designed to serve as a docking tag, was fused to the N-terminus of human vascular endothelial growth factor that served as a targeting protein. An 18-125 and an 18-127 amino acid long fragments of RNase I were engineered, expressed and refolded into active conformations to serve as adapter proteins. Interactions between the targeting and adapter proteins were characterized using enzymatic analysis and surface plasmon resonance. Targeting DNA delivery complexes were assembled, characterized by dynamic light scattering, and found to be very effective in receptor-mediated DNA delivery. PMID- 12737853 TI - SARS: experience at Prince of Wales Hospital, Hong Kong. PMID- 12737855 TI - Towards clinical transplantation tolerance. PMID- 12737854 TI - The dietary fibre debate: more food for thought. PMID- 12737856 TI - Death in anorexia nervosa. PMID- 12737857 TI - Dietary fibre and colorectal adenoma in a colorectal cancer early detection programme. AB - BACKGROUND: Although dietary fibre has been reported to have no association with colorectal adenoma and cancer, in some studies this topic remains controversial. METHODS: We used a 137-item food frequency questionnaire to assess the relation of fibre intake and frequency of colorectal adenoma. The study was done within the Prostate, Lung, Colorectal, and Ovarian (PLCO) Cancer Screening Trial, a randomised controlled trial designed to investigate methods for early detection of cancer. In our analysis, we compared fibre intake of 33971 participants who were sigmoidoscopy-negative for polyps, with 3591 cases with at least one histologically verified adenoma in the distal large bowel (ie, descending colon, sigmoid colon, or rectum). Odds ratios were estimated by logistic regression analysis. FINDINGS: High intakes of dietary fibre were associated with a lower risk of colorectal adenoma, after adjustment for potential dietary and non dietary risk factors. Participants in the highest quintile of dietary fibre intake had a 27% (95% CI 14-38, p(trend)=0.002) lower risk of adenoma than those in the lowest quintile. The inverse association was strongest for fibre from grains and cereals and from fruits. Risks were similar for advanced and non advanced adenoma. Risk of rectal adenoma was not significantly associated with fibre intake. INTERPRETATION: Dietary fibre, particularly from grains, cereals, and fruits, was associated with decreased risk of distal colon adenoma. PMID- 12737858 TI - Dietary fibre in food and protection against colorectal cancer in the European Prospective Investigation into Cancer and Nutrition (EPIC): an observational study. AB - BACKGROUND: Dietary fibre is thought to protect against colorectal cancer but this view has been challenged by recent prospective and intervention studies that showed no protective effect. METHODS: We prospectively examined the association between dietary fibre intake and incidence of colorectal cancer in 519978 individuals aged 25-70 years taking part in the EPIC study, recruited from ten European countries. Participants completed a dietary questionnaire in 1992-98 and were followed up for cancer incidence. Relative risk estimates were obtained from fibre intake, categorised by sex-specific, cohort-wide quintiles, and from linear models relating the hazard ratio to fibre intake expressed as a continuous variable. FINDINGS: Follow-up consisted of 1939011 person-years, and data for 1065 reported cases of colorectal cancer were included in the analysis. Dietary fibre in foods was inversely related to incidence of large bowel cancer (adjusted relative risk 0.75 [95% CI 0.59-0.95] for the highest versus lowest quintile of intake), the protective effect being greatest for the left side of the colon, and least for the rectum. After calibration with more detailed dietary data, the adjusted relative risk for the highest versus lowest quintile of fibre from food intake was 0.58 (0.41-0.85). No food source of fibre was significantly more protective than others, and non-food supplement sources of fibre were not investigated. INTERPRETATION: In populations with low average intake of dietary fibre, an approximate doubling of total fibre intake from foods could reduce the risk of colorectal cancer by 40%. PMID- 12737860 TI - The aggressive treatment of hypertension. PMID- 12737859 TI - Tolerogenic immunosuppression for organ transplantation. AB - BACKGROUND: Insight into the mechanisms of organ engraftment and acquired tolerance has made it possible to facilitate these mechanisms, by tailoring the timing and dosage of immunosuppression in accordance with two therapeutic principles: recipient pretreatment, and minimum use of post-transplant immunosuppression. We aimed to apply these principles in recipients of renal and extrarenal organ transplants. METHODS: 82 patients awaiting kidney, liver, pancreas, or intestinal transplantation were pretreated with about 5 mg/kg of a broadly reacting rabbit antithymocyte globulin during several hours. Post transplant immunosuppression was restricted to tacrolimus unless additional drugs were needed to treat breakthrough rejection. After 4 months, patients on tacrolimus monotherapy were considered for dose-spacing to every other day or longer intervals. FINDINGS: We frequently saw evidence of immune activation in graft biopsy samples, but unless this was associated with graft dysfunction or serious immune destruction, treatment usually was not intensified. Immunosuppression-related morbidity was virtually eliminated. 78 (95%) of 82 patients survived at 1 year and at 13-18 months. Graft survival was 73 (89%) of 82 at 1 year and 72 (88%) of 82 at 13-18 months. Of the 72 recipients with surviving grafts, 43 are on spaced doses of tacrolimus monotherapy: every other day (n=6), three times per week (11), twice per week (15), or once per week (11). INTERPRETATION: The striking ability to wean immunosuppression in these recipients indicates variable induction of tolerance. The simple therapeutic principles are neither drug-specific nor organ-specific. Systematic application of these principles should allow improvements in quality of life and long-term survival after organ transplantation. PMID- 12737861 TI - Endothelial dysfunction and raised plasma concentrations of asymmetric dimethylarginine in pregnant women who subsequently develop pre-eclampsia. AB - BACKGROUND: Maternal endothelial dysfunction is a feature of established pre eclampsia but whether this is a cause or consequence of the disorder is not clear. We tested the hypothesis that endothelial dysfunction and raised plasma concentrations of asymmetric dimethylarginine (ADMA), the endogenous inhibitor of endothelial nitric oxide synthase, precede and contribute to the development of pre-eclampsia. METHODS: We assessed uterine artery doppler waveforms in 86 women at 23-25 weeks' gestation. We tested endothelial function in all women using flow mediated dilation of the brachial artery at 23-25 weeks' gestation. Plasma concentrations of ADMA were also measured. FINDINGS: 43 women had normal uterine artery doppler waveforms and subsequently had a normal outcome. The second group of 43 had evidence of impaired placental perfusion and of these, 19 (44%) had normal outcome, 14 (33%) developed intrauterine growth restriction of the fetus (IUGR), and pre-eclampsia arose in ten (23%). Women who developed pre-eclampsia had significantly lower flow-mediated dilation than did women who had normal outcome (3.58% [SD 2.76] vs 8.59% [2.76]; p<0.0001). Irrespective of pregnancy outcome, women with evidence of impaired placental perfusion had significantly higher levels of ADMA than did women with normal doppler waveforms (2.4 micromol/L [IQR 1.97-3.14] vs 0.81 micromol/L [0.49-1.08]; p<0.0001). There was a strong inverse correlation between ADMA and flow-mediated dilation but only in the group of women who eventually developed pre-eclampsia (r=-0.8, p=0.005). INTERPRETATION: Maternal endothelial function is impaired in women who eventually develop pre-eclampsia and it occurs before the development of the clinical syndrome. Furthermore, women with high resistance placental circulation at risk of pre-eclampsia, IUGR, or both have raised concentrations of ADMA, which is a potential contributory factor for pre-eclampsia, and is associated with endothelial dysfunction in some women. PMID- 12737863 TI - A black necrotic ulcer. PMID- 12737864 TI - Effectiveness of precautions against droplets and contact in prevention of nosocomial transmission of severe acute respiratory syndrome (SARS). AB - We did a case-control study in five Hong Kong hospitals, with 241 non-infected and 13 infected staff with documented exposures to 11 index patients with severe acute respiratory syndrome (SARS) during patient care. All participants were surveyed about use of mask, gloves, gowns, and hand-washing, as recommended under droplets and contact precautions when caring for index patients with SARS. 69 staff who reported use of all four measures were not infected, whereas all infected staff had omitted at least one measure (p=0.0224). Fewer staff who wore masks (p=0.0001), gowns (p=0.006), and washed their hands (p=0.047) became infected compared with those who didn't, but stepwise logistic regression was significant only for masks (p=0.011). Practice of droplets precaution and contact precaution is adequate in significantly reducing the risk of infection after exposures to patients with SARS. The protective role of the mask suggests that in hospitals, infection is transmitted by droplets. PMID- 12737865 TI - Haemorrhagic-fever-like changes and normal chest radiograph in a doctor with SARS. AB - A 33-year-old doctor contracted severe acute respiratory syndrome presenting with features of disseminated intravascular coagulopathy without changes in the chest radiograph initially. A CT scan of his chest showed marked lung changes. His condition improved with intravenous methylprednisolone 500 mg daily and ribavirin 1.2 g orally thrice daily. The case illustrates the importance of a break in fever between the viraemic and lung inflammatory phases of the illness that occurs before radiographic changes and which may obscure diagnosis. Careful quarantine and follow-up of these patients are necessary. Coagulopathy is usually uncomplicated and early CT of the chest may elucidate hidden lung changes and facilitate a rapid diagnosis. PMID- 12737866 TI - Risk of vaccine failure after Haemophilus influenzae type b (Hib) combination vaccines with acellular pertussis. AB - An increase in invasive Hib disease incidence in the UK has coincided with the distribution of combination vaccines that contain acellular pertussis (DTaP-Hib). These vaccines have been associated with reduced immunogenicity of the Hib component, although there is little agreement on the clinical relevance of this finding. We retrospectively compared vaccine formulations given to fully vaccinated Hib cases with those administered to fully immunised age-matched controls using conditional logistic regression. More cases than controls received all three doses of their infant primary course as DTaP-Hib, compared with two or three doses of another Hib vaccine (conditional odds ratio 6.77 [95% CI 3.26 14.07]). PMID- 12737868 TI - Canada fury at WHO ban. PMID- 12737869 TI - ASEAN leaders agree regional SARS plan. PMID- 12737867 TI - Antibody to Haemophilus influenzae type b after routine and catch-up vaccination. AB - Since 1999, the number of cases of Haemophilus influenzae type b (Hib) disease in the UK has risen. We investigated the role of population immunity in this change by testing more than 2600 serum samples from children aged 1-15 years. After the introduction of the routine Hib conjugate vaccination programme for infants, median antibody titres rose significantly in 1-year-olds. Individuals who received their first dose of vaccine at age 1-4 years in the original catch-up campaign initially had much higher concentrations of antibody than those who had been immunised in infancy. A second catch-up campaign in children aged 6 months to 4 years should be highly effective in boosting immunity and reducing disease in the short term. PMID- 12737870 TI - Spreading the word about HIV/AIDS in India. PMID- 12737871 TI - Wnt helps blood stem cells tick over. PMID- 12737872 TI - US researcher broke federal rules in aiding Chinese HIV study. PMID- 12737873 TI - US Veterans Administration research under spotlight. PMID- 12737875 TI - Money being wasted in fight against malaria, says MSF. PMID- 12737877 TI - Panel advises against egg and sperm donations from siblings. PMID- 12737878 TI - Primary pulmonary hypertension. AB - Primary pulmonary hypertension (PPH) is a rare disorder characterised by raised pulmonary-artery pressure in the absence of secondary causes. Precapillary pulmonary arteries are affected by medial hypertrophy, intimal fibrosis, microthrombosis, and plexiform lesions. Most individuals present with dyspnoea or evidence of right heart failure. Echocardiography is the best non-invasive test to screen for suspected pulmonary hypertension. The discovery of mutations in the coding region of the gene for bone morphogenetic protein receptor 2 in patients with familial and sporadic PPH may help not only to elucidate pathogenesis but also to direct future treatment options. The pathogenesis is not completely understood, but recent investigations have revealed many possible candidate modifier genes. Without treatment, the disorder progresses in most cases to right heart failure and death. With current therapies such as epoprostenol, progression of disease is slowed, but not halted. Many promising new therapeutic options, including prostacyclin analogues, endothelin-1-receptor antagonists, and phosphodiesterase inhibitors, improve clinical function and haemodynamic measures and may prolong survival. PMID- 12737879 TI - Diabetic foot ulcers. AB - Ulceration of the foot in diabetes is common and disabling and frequently leads to amputation of the leg. Mortality is high and healed ulcers often recur. The pathogenesis of foot ulceration is complex, clinical presentation variable, and management requires early expert assessment. Interventions should be directed at infection, peripheral ischaemia, and abnormal pressure loading caused by peripheral neuropathy and limited joint mobility. Despite treatment, ulcers readily become chronic wounds. Diabetic foot ulcers have been neglected in health care research and planning, and clinical practice is based more on opinion than scientific fact. Furthermore, the pathological processes are poorly understood and poorly taught and communication between the many specialties involved is disjointed and insensitive to the needs of patients. PMID- 12737881 TI - Overcoming obstacles in confronting torture. PMID- 12737880 TI - Von Recklinghausen's neurofibromatosis: neurofibromatosis type 1. PMID- 12737882 TI - The medical community's response to torture. PMID- 12737883 TI - Visiting detainees to prevent torture. PMID- 12737884 TI - Aminopeptidase N inhibitors and SARS. PMID- 12737885 TI - Effect of ibuprofen on cardioprotective effect of aspirin. PMID- 12737886 TI - Effect of ibuprofen on cardioprotective effect of aspirin. PMID- 12737887 TI - Effect of ibuprofen on cardioprotective effect of aspirin. PMID- 12737889 TI - Effect of ibuprofen on cardioprotective effect of aspirin. PMID- 12737890 TI - Effect of ibuprofen on cardioprotective effect of aspirin. PMID- 12737891 TI - Effect of ibuprofen on cardioprotective effect of aspirin. PMID- 12737893 TI - Women's health and human rights in HIV prevention research. PMID- 12737895 TI - Women's health and human rights in HIV prevention research. PMID- 12737894 TI - Infections related to central venous catheters in US intensive-care units. PMID- 12737896 TI - Benign prostatic hyperplasia and sexual dysfunction. PMID- 12737897 TI - Benign prostatic hyperplasia and sexual dysfunction. PMID- 12737898 TI - Return of Haemophilus influenzae type b infections. PMID- 12737899 TI - Return of Haemophilus influenzae type b infections. PMID- 12737900 TI - Return of Haemophilus influenzae type b infections. PMID- 12737901 TI - Bacterial meningitis in Angola. PMID- 12737902 TI - Simian virus 40 infection in lymphoproliferative disorders. PMID- 12737904 TI - 15 or 33 years of cord-blood transplantation? PMID- 12737905 TI - Tighter regulation needed for dietary supplements in USA. PMID- 12737906 TI - Brain natriuretic peptide as a predictor of cardiomyopathy in Chagas' disease. PMID- 12737907 TI - Trial and error. PMID- 12737908 TI - The death of academic clinical trials. PMID- 12737909 TI - Home-based model for HIV voluntary counselling and testing. PMID- 12737910 TI - More money for AIDS. PMID- 12737912 TI - Too much talk in theatre. PMID- 12737913 TI - Screening for diabetic retinopathy. PMID- 12737914 TI - Screening for diabetic retinopathy. PMID- 12737915 TI - Danielle Ofri. PMID- 12737920 TI - Interface pressure data and the prediction of driver discomfort in road trials. AB - An objective measure that will predict discomfort reliably, and which can be used at an early stage in the development of a vehicle and its seating, would have the potential to reduce the prevalence of musculoskeletal problems associated with driving. This paper reports on an extended road trial study to further investigate the potential value of pressure distribution data in the prediction of reported discomfort. Road trial data were collected from three cars and then interface pressure data were recorded for each of the three seats. Clear differences were identified between the cars with respect to reports of discomfort. However, no clear relationship was found between interface pressure data and reported discomfort. PMID- 12737921 TI - Maximum acceptable weights for asymmetric lifting of Chinese females. AB - This study used the psychophysical approach to evaluate the effects of asymmetric lifting on the maximum acceptable weight of lift (MAWL) and the resulting heart rate, oxygen uptake and rating of perceived exertion (RPE). A randomized complete block factorial design was employed. Twelve female college students lifted weights at three different lifting frequencies (one-time maximum, 1 and 4 lifts/min) in the sagittal plane and at three different asymmetric angles (30 degrees, 60 degrees, and 90 degrees ) from the floor to a 68-cm height pallet. This lifting experiment was conducted for a 1-h work period using a free-style lifting technique. The MAWLs for asymmetric lifting were significantly lower than those for symmetric lifting in the sagittal plane. The MAWL decreased with the increase in the angle of asymmetry. However, the heart rate, oxygen uptake and RPE remained unchanged. Though the MAWL decreased significantly with lifting frequency, both the physiological costs (heart rate and oxygen uptake) and rating of perceived exertion increased with the increase in lift frequency. The most stressed body part was the arm. Lifting frequency had no significant effect on the percentage decrease in MAWL from the sagittal plane values. On average, decreases of 5%, 9% and 14% for MAWL at 30 degrees, 60 degrees and 90 degrees asymmetric lifting, respectively, were revealed. This result was in agreement with the findings of Chinese males studied by Wu [Int. J. Ind. Ergonom. 25 (2000) 675]. The percentage decrease in MAWL with twisting angle for the Chinese participants was somewhat lower than those for Occidental participants. In addition, even though there was an increase in heart rate and RPE with the increase in the symmetrical lift angle for Occidental participants, it was different from the Chinese participants. Lastly, the 1991 NIOSH equation asymmetry multiplier is more conservative in comparison with the results of the present study. PMID- 12737922 TI - The effect of technique and shaft configuration in snow shoveling on physiologic, kinematic, kinetic and productivity variables. AB - A repeated measures 2 x 2 factorial design using a psychophysical experimental methodology was performed to quantify the effect of shaft design (straight and bent shaft) and shoveling technique (forward and backward progression) on heart rate, perceived exertion, productivity, trunk kinematics and load kinetics. Ten male subjects performed four 8-min trials of snow shoveling on a paved asphalt surface. The most notable finding was significantly less trunk flexion with the bent shaft (41.4 degrees ) than with the straight shaft design (49.2 degrees ). The study results led to a recommendation of a bent-shaft shovel for the purpose of reducing trunk flexion. In the absence of any strong positive evidence and due to poor subjective response to backward progression while shoveling, this technique was not recommended. PMID- 12737923 TI - Failure to adapt or adaptations that fail: contrasting models on procedures and safety. AB - This paper introduces two models on procedures and safety and assesses the practical consequences these have for organizations trying to make progress on safety through procedures. The application of procedures is contrasted as rote rule following versus substantive cognitive activity. It reveals a fundamental double bind: operators can fail to adapt procedures when adapting proved necessary, or attempt procedural adaptations that may fail. Rather than simply increasing pressure to comply, organizations should invest in their understanding of the gap between procedures and practice, and help develop operators' skill at adapting. PMID- 12737924 TI - Computer supported visualisation of quality systems developed by network teams. AB - This paper reports a pilot study and three case studies to examine aimed at studying if a quality system, according to the ISO 9000 quality standard, can be visualised, exploring how the visualisation should be designed in order to support continuous improvements and evaluating the design process for development of prototypes. By discussing with the research team, three companies in southern Sweden set up design teams to establish principles for visualisations of their quality systems on their intranets. Together with one of the researchers, the design teams created and evaluated computer supported prototypes and exchanged ideas between the teams via the Internet. The results show that quality systems can be adequately visualised and that the companies preferred descriptions of the physical plant layout containing symbols that connect to further information by hyperlinks. A quality system based on computer supported visualisation will make the quality system more understandable and have better possibilities to engage personnel in the quality work; it will be faster, easier, and more interesting to use than systems with only paper documents. The use of design teams with access to each other's prototypes via the Internet supported the design process by stimulating generation of ideas and solutions to visualise a quality system. PMID- 12737925 TI - Evaluation of driver discomfort during long-duration car driving. AB - The evolution of indices of fatigue, discomfort, and performance of subjects seated for long duration (150 min) in car seats were studied (n=11). Four experimental configurations were used: with and without vibration for two seats (U, uncomfortable; C, comfortable). Surface electromyography (SEMG) data were recorded bilaterally from cervical erector spinae and external oblique muscles. Discomfort increased significantly during the trial, regardless of the experimental condition (p<0.05). Performance was significantly worse for seat U with vibration (p<0.05). The median frequency of SEMG signals did not change between experimental conditions or across time. It would appear that, either the level of discomfort experienced was insufficient to change either performance or SEMG measures, or that the large parameter estimation variance of the SEMG signals might have masked any underlying spectral change. Further refinement of the SEMG signal processing methodology may be necessary to be able to detect fatigue of postural muscles. PMID- 12737926 TI - Comparison of four different backpacks intended for school use. AB - Four backpacks were evaluated for their desireability for use as school bags. Three of the four backpacks were specifically designed for school use based on previous research and ergonomic principles while the fourth (standard) backpack was chosen from two backpacks that their manufacturer considered to be the most likely to be used as a school bag. Twelve school students evaluated each of the backpacks firstly by examining them, again after donning them and again after walking with them on a treadmill by completing a questionnaire asking about the appearance, function and comfort of each backpack. On initial examination, the standard backpack was the most favoured but as functionality became increasingly important during the treadmill walk, the backpack which was designed specifically for school use and had two major compartments, substantial back padding and side compression straps became the most favoured. This particular design of backpack was reported as having the greatest practicality, being the least physically demanding and allowing the greatest balance and ease of walking. The results of this study suggest that school student's preference of backpack may change from when they first examine a prospective backpack to when they have used it. The study also shows that school students' preferred attributes in a backpack may shift over this time from 'style and image' to 'function and fit'. PMID- 12737927 TI - Aspects to improve cabin comfort of wheel loaders and excavators according to operators. AB - Comfort plays an increasingly important role in interior design of earth moving equipment. Although research has been conducted on vehicle interiors of wheel loaders and excavators, hardly any information is known about the operator's opinion. In this study a questionnaire was completed by machine operators to get their opinion about aspects which need to be improved in order to design a more comfortable vehicle interior. The results show that almost half of the operators rate the comfort of their cabin "average" or "poor". According to the operators, cab comfort of wheel loaders can be increased by improving seat comfort. Besides improving seat comfort, cabin comfort of excavators can be improved by changing the cab design (including dimensions, ingress/egress), view, reliability, and climate control. PMID- 12737928 TI - The 3D scanner for measuring body surface area: a simplified calculation in the Chinese adult. AB - Three-dimensional (3D) surface anthropometry enables us to extend the study to 3D geometry and morphology of mainly external human body tissues. A model is presented for estimation of human body surface area (BSA), which is identical in form to the one proposed in 1916 by DuBois and DuBois is presented. The purpose of this study is to measure BSA, using 3D scanner, and to derive a simple BSA estimation formula for the Chinese adults. In as little as 12s, the Chang Gung Whole-Body Scanner (CGWBS) allows you to capture the shape of the entire human body. The total error in BSA measurement due to scanning measurement and software computational error is less than 1%. The 3D anthropometric measures in a healthy population (n=3951) were investigated, and the results were used to derive a BSA estimation formula. The results seem to be comparable to previous data that measured BSA using traditional methods. The BSA estimation model of this study also validated using 300 new measurements along with the formulae proposed in previous researches. The result suggests that our formula better fits our adults. PMID- 12737929 TI - Human vestibular memory studied via measurement of the subjective horizontal during gondola centrifugation. AB - Measurements of the subjective visual horizontal (SVH) were made in a large swing out gondola centrifuge. Rotation of the centrifuge was anti-clockwise, as seen from above. Test subjects were seated upright in the gondola, facing forwards. In front of the subject, at a straight-ahead eye-level position, there was a narrow luminous line, which could be rotated, by remote control, about the visual axis. At gravitoinertial force levels of 1.1-1.3G the subjects were asked to indicate, by repeatedly setting the line in darkness, what they perceived as horizontal (the SVH). During gondola centrifugation, the head and body length axis is always parallel with the resultant gravitoinertial force vector (vectorial sum of earth gravity force and the centrifugal force) i.e., the horizontal plane of the head or body does not change with respect to the gravitoinertial horizontal. Hence, the otolith organs, as well as the somatosensory system, continually signal upright position. However, the swing-out of the gondola during acceleration of the centrifuge (25 degrees at 1.1G) is a roll (frontal plane) change-in-position stimulus to the vertical semicircular canals, thus creating an otolith semicircular canal conflict. After acceleration of the centrifuge, the SVH was initially tilted up to 20 degrees to the right relative to the gravitoinertial horizontal. Since there was no roll-tilt stimulus to gravity receptors, this SVH tilt must be related to stimulation of the semicircular canals. However, it decayed much more slowly than any known effects of angular-velocity stimulation of the semicircular canals. The decay was bi-phasic with two time constants, the smaller in the region of 1-2 min, the other being too large to be reliably estimated on the basis of data collected during only 10 min. This persistence of the SVH tilt suggests a memory for angular changes in roll head position detected by the semicircular canals-a position-storage mechanism. Further, the SVH seems to be dependent on two different mechanisms related to semicircular canal stimulation. PMID- 12737930 TI - Chronic administration of corticosterone impairs spatial reference memory before spatial working memory in rats. AB - Corticosterone (CORT), the predominant glucocorticoid in rodents, elevated for 21 days damages hippocampal subregion CA3. We tested the hypothesis that CORT would impair spatial memory, a hippocampal function. In each of the three experiments, rats received daily, subcutaneous injections of either CORT (26.8 mg/kg body weight in sesame oil) or sesame oil vehicle alone (VEH). CORT given for 21 or 56 days effectively attenuated body weight gain and reduced selective organ and muscle weights. All behavioral testing was done on tasks that are minimally stressful and avoid deprivation. For each experiment, testing commenced 24h after the last injection. CORT given for 21 days did not impair spatial working memory in the Y-maze (Experiments 1 and 2). After 56-day administration of CORT, spatial working memory was impaired in the Y-maze (Experiment 2). CORT given for 21 days also failed to impair spatial working memory in the Barnes maze (Experiment 3). However, in trials that depended solely on reference memory, the VEH group improved in performance, whereas the CORT group did not. In conclusion, CORT elevated over a period of 21 days did not impair spatial working memory, but impaired the formation of a longer-term form of memory, most likely reference memory. Impairments in spatial working memory are seen only after longer durations of CORT administration. PMID- 12737931 TI - Bilateral lesions of the insular cortex or of the prefrontal cortex block the association between taste and odor in the rat. AB - The neural basis for the association between taste and odor was investigated in the rat. First, behavioral procedures to study the mechanisms underlying the association between qualitative aspects of the odor and taste in rats were developed. Rats were presented with several pairings of the 0.3 mol/L NaCl solution and a flavor, and pairings of distilled water and another flavor. Then the rats received an IP injection of a furosemide to develop sodium deficiency. On the next day, the rats were presented with either of two types of odor flavored water: water in which the flavor had been paired with NaCl, or water in which the flavor (grape or coffee) had been paired with distilled water. Normal rats avoided ingesting the water flavored with the odor previously paired with NaCl. Sodium-deprived rats, however, ingested the water flavored with that odor. Rats with lesions in either the insular cortex or in the prefrontal cortex neither preferred nor avoided the water flavored with the odor paired with NaCl. It was concluded that rats acquire association between taste and odor, and that the insular and the prefrontal cortices of the rats were involved in this association. PMID- 12737932 TI - Involvement of the hippocampal CA3-region in acquisition and in memory consolidation of spatial but not in object information in mice. AB - This study investigates the implication of the hippocampal CA3-region in the different phases of learning and memory in spatial and non-spatial tasks. For that purpose, we performed focal injections of diethyldithiocarbamate (DDC) into the CA3-region of the dorsal hippocampus. The DDC chelates most of the heavy metals in the brain which blocks selectively and reversibly the synapses containing heavy metals, i.e., the mossy fibres synaptic buttons and synapses of the dendrites of pyramidal cells. The effects of temporal inactivation of the CA3 region was examined in a non-associative task, the spatial open-field, designed to estimate the ability of mice to react to spatial changes, and in the object recognition task, designed to estimate the ability of mice to identify a familiar object. The results show that DDC induced a specific impairment on learning and memory consolidation in the spatial open-field but had no effect on recall in this task. In the object recognition task, DDC did not induce any impairment in the different phases of learning and memory. These data demonstrate that the hippocampal CA3-region is specifically implicated in spatial information processing and seems to be involved not only in acquisition but also in consolidation of spatial information. PMID- 12737933 TI - Central galanin administration blocks consolidation of spatial learning. AB - Galanin is a neuropeptide that inhibits the evoked release of several neurotransmitters, inhibits the activation of intracellular second messengers, and produces deficits in a variety of rodent learning and memory tasks. To evaluate the actions of galanin on encoding, consolidation, and storage/retrieval, galanin was acutely administered to Sprague-Dawley rats at time points before and after training trials in the Morris water maze. Intraventricular administration of galanin up to 3h after subjects had completed daily training trials in the Morris water task impaired performance on the probe trial, indicating that galanin-blocked consolidation. Pretreatment with an adenylate cyclase activator, forskolin, prevented the deficits in distal cue learning produced by galanin. Di-deoxyforskolin, an inactive analog of forskolin, had no effect. These results provide the first evidence that galanin interferes with long-term memory consolidation processes. A potential mechanism by which galanin produces this impairment may involve the inhibition of adenylate cyclase activity, leading to inhibition of downstream molecular events that are necessary for consolidation of long-term memory. PMID- 12737934 TI - Perinatal exposure to diethylstilbestrol improves olfactory discrimination learning in male and female Swiss-Webster mice. AB - During late prenatal and early postnatal brain development, estrogen induces structural sex differences that correspond to behavioral differences in certain domains such as learning and memory. The typically superior performance of males is attributed to the action of elevated concentrations of estrogen, derived inside neurons from the aromatization of testosterone. In contrast, female performance appears dependent on minimal estrogenic activity. Rat models of the relationship between hormones and cognitive behavior predominate the field, but the advent of genetically modified mice as research tools necessitates development of analogous mouse models. This study examined how early postnatal exposure to the synthetic estrogen diethylstilbestrol (DES) affected the ability of male and female Swiss-Webster mice to learn a two-choice olfactory discrimination and three repeated reversals. Mice treated with subcutaneous injections of DES from postnatal days 1-10 learned reversals more readily than oil-treated controls, a difference that became evident after repeated testing. DES-exposed males and females learned reversals at a comparable rate, suggesting that early postnatal estrogen exposure does not influence this mode of learning through a sexually differentiated mechanism in mice. An analysis of response patterns during qualitatively different phases of reversal learning revealed that DES-induced improvements probably were not due to greater inhibitory control. Instead, DES appeared to enhance associative ability. Early postnatal estrogen exposure may have the potential to preserve certain cognitive skills in adulthood. PMID- 12737935 TI - Combined uridine and choline administration improves cognitive deficits in spontaneously hypertensive rats. AB - Rationale. Hypertension is considered a risk factor for the development of cognitive disorders, because of its negative effects on cerebral vasculature and blood flow. Genetically induced hypertension in rats has been associated with a range of cognitive impairments. Therefore, spontaneously hypertensive rats (SHR) can potentially be used as a model for cognitive deficits in human subjects. Consecutively, it can be determined whether certain food components can improve cognition in these rats. Objective. The present study aimed to determine whether SHR display specific deficits in attention, learning, and memory function. Additionally, effects of chronic uridine and choline administration were studied. Methods. 5-7 months old SHR were compared with normotensive Wistar-Kyoto (WKY) and Sprague-Dawley (SD) rats. (a) The operant delayed non-matching-to-position (DNMTP) test was used to study short-term memory function. (b) The five-choice serial reaction time (5-CSRT) task was used to assess selective visual attention processes. (c) Finally, the Morris water maze (MWM) acquisition was used as a measure for spatial learning and mnemonic capabilities. Results. (1) SHR exhibited significantly impaired performance in the 5-CSRT test in comparison with the two other rat strains. Both the SHR and WKY showed deficits in spatial learning when compared with the SD rats. (2) Uridine and choline supplementation normalized performance of SHR in the 5-CSRT test. (3) In addition, uridine and choline treatment improved MWM acquisition in both WKY and SHR rats. Conclusion. The present results show that the SHR have a deficiency in visual selective attention and spatial learning. Therefore, the SHR may provide an interesting model in the screening of substances with therapeutic potential for treatment of cognitive disorders. A combination of uridine and choline administration improved selective attention and spatial learning in SHR. PMID- 12737936 TI - Hippocampal gene expression profiling in spatial discrimination learning. AB - Learning and long-term memory are thought to involve temporally defined changes in gene expression that lead to the strengthening of synaptic connections in selected brain regions. We used cDNA microarrays to study hippocampal gene expression in animals trained in a spatial discrimination-learning paradigm. Our analysis identified 19 genes that showed statistically significant changes in expression when comparing Nai;ve versus Trained animals. We confirmed the changes in expression for the genes encoding the nuclear protein prothymosin(alpha) and the delta-1 opioid receptor (DOR1) by Northern blotting or in situ hybridization. In additional studies, laser-capture microdissection (LCM) allowed us to obtain enriched neuronal populations from the dentate gyrus, CA1, and CA3 subregions of the hippocampus from Nai;ve, Pseudotrained, and spatially Trained animals. Real time PCR examined the spatial learning specificity of hippocampal modulation of the genes encoding protein kinase B (PKB, also known as Akt), protein kinase C(delta) (PKC(delta)), cell adhesion kinase(beta) (CAK(beta), also known as Pyk2), and receptor protein tyrosine phosphatase(zeta/beta) (RPTP(zeta/beta)). These studies showed subregion specificity of spatial learning-induced changes in gene expression within the hippocampus, a feature that was particular to each gene studied. We suggest that statistically valid gene expression profiles generated with cDNA microarrays may provide important insights as to the cellular and molecular events subserving learning and memory processes in the brain. PMID- 12737937 TI - Phenotypic expression of the C282Y/Q283P compound heterozygosity in HFE and molecular modeling of the Q283P mutation effect. AB - In Caucasians, from 4 to 35% of hereditary hemochromatosis (HH) patients carry a least one chromosome without a common assigned HFE mutation (i.e., C282Y, H63D, and S65C). We have undertaken a D-HPLC scanning of the HFE coding region in such patients in order to identify uncommon mutations liable to explain their high transferrin saturation level. Twenty HH patients from Brittany carrying at least one chromosome without an assigned mutation were selected on the basis of a transferrin saturation level with the following threshold: > or = 60% in men and > or = 50% in women, in the absence of other known causes of iron disorders. This strategy allowed us to detect a heterozygous sequence variant in exon 4 of the HFE gene from one individual who was also heterozygous for C282Y. Subsequent DNA sequencing analysis identified an adenine to cytosine transversion at position 848 which changes amino acid 283 from glutamine to proline (Q283P). Family study revealed a clear association between the C282Y/Q283P compound heterozygote genotype and the development of HH. Molecular modeling studies are in favor of a destabilizing effect of the Q283P mutation on the tertiary structure of the HFE protein. This is the first report of a natural protein variant describing the introduction of a proline in a central beta-strand position. Our approach may have practical implications in screening strategies for hereditary hemochromatosis, molecular diagnosis, and HFE structure-function relationships. PMID- 12737938 TI - Mild hemolysis in a girl with G6PD Sumare (class I variant) associated with G6PD A-. AB - In the present study we describe the clinical and laboratory features of a female child, a compound heterozygote for glucose-6-phosphate dehydrogenase (G6PD) Sumare (1292T-->G) and African variants (202G-->A). G6PD Sumare is a variant causing chronic nonspherocytic hemolytic anemia. The child had neonatal jaundice 2 days after birth and needed phototherapy for 8 days. Since then, she has not had episodes of dark urine or new episodes of jaundice. She has not had hemolytic crises in spite of five respiratory infections and antibiotics administration. Laboratory data showed a reticulocytosis (5.6%) without anemia and serum unconjugated bilirubin at the upper limit of the normalcy. No hemoglobin and hemosiderin in the urine were detected. G6PD activity at 37 degrees C was 1.15 UI/g Hb and G6PD cellulose acetate electrophoresis at pH 9.0 revealed two bands, in equal amounts, with normal and faster migration, respectively. She was homozygous for the normal (TA)6(TA)6 repeat in the UGT1A1 promoter. We conclude that the association of G6PD Sumare and G6PD A- gave rise to a very mild chronic hemolysis, and the red cell population containing G6PD A- is probably enough to protect against severe chronic hemolysis. PMID- 12737939 TI - Glycoprotein IIb/IIIa complex is the target in mirtazapine-induced immune thrombocytopenia. AB - Mirtazapine (MW 265.36), a tetracyclic antidepressant of the piperazine-azapine group which augments central noradrenergic and serotonergic activity, is currently used as an oral antidepressant. We report a case of severe thrombocytopenia in a 66-year-old patient occurring after mirtazapine administration, suggesting an immune mechanism. This report documents the first case of mirtazapine-induced immune thrombocytopenia. The patient's serum was screened for drug-induced anti-platelet antibody with the chromium(51) (Cr(51)) platelet lysis technique. The drug-dependent antibody was characterized using flow cytometry, the monoclonal antibody immobilization of platelet antigens assay (MAIPA assay), and immunoprecipitation. By the Cr(51) platelet lysis technique, we obtained an equivocal result for the detection of mirtazapine-induced antibody. However, the patient's serum tested positive for mirtazapine-induced antibody by flow cytometry. The results showed that the binding ratio of 5.7 (mean fluorescence intensity) in the presence of the patient's serum and mirtazapine in a final concentration of 1.0 mmol/L was strongly positive. The antibody was found to bind the glycoprotein (GP) IIb/IIIa complex by MAIPA assay by using five different monoclonal antibodies against GP complexes Ib/IX, GPIIb/IIIa, or GPIa/IIa. Immunoprecipitation studies showed that the GPIIb/IIIa complex was precipitated by antibody in the presence, but not in the absence, of mirtazapine. These findings provide evidence that immune thrombocytopenia can be caused by sensitivity to the antidepressant mirtazapine. This is the first well documented case of mirtazapine-induced immune thrombocytopenia. PMID- 12737940 TI - Molecular characterization of G6PD Insuli--a novel 989 CGC --> CAC (330 Arg --> His) mutation in the Indian population. AB - Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common red cell enzymopathy among humans. G6PD deficiency was reported in India more than 30 years ago and about 13 biochemically characterized variants have been reported. We now describe the molecular characterization of a distinct biochemical variant from India which was previously characterized by us as a unique Class IV variant with fast electrophoretic mobility. The sequence of the G6PD gene from this variant has revealed the presence of a novel 989 G --> A mutation in exon 9 with a predicted amino acid change of Arg330His. PMID- 12737941 TI - Interaction of erythroid spectrin with hemoglobin variants: implications in beta thalassemia. AB - Among the few studies, producing contradictory results, done on the interaction of erythroid membrane skeletal spectrin with hemoglobin (Hb), none has been able to provide a quantitative estimate of the association of spectrin with Hb. In this work, studies on the interactions of erythroid spectrin with Hb have been elaborated upon using a novel fluorescence technique. The concentration-dependent change in the fluorescence intensity of fluorescein-conjugated spectrin (F spectrin) in presence of oxy-Hb indicated binding with a dissociation constant of approximately 20 microM that has been directly evaluated from the increase in the extent of quenching of the fluorescein fluorescence of F-spectrin by reverse titration with the increasing concentrations of different Hb samples isolated from both normal and beta-thalassemic patients. The Hb compositions, with major components of the normal HbA, the fetal HbF, and the variant HbA2, of each individual were estimated using the Variant HPLC device of Bio-Rad. Results of the present study indicated that the dissociation constant, K(d), of spectrin binding to Hb decreased from 19.5 +/- 2 microM in normal individuals to of 6.5 +/ 0.5 microM in the presence of 73% HbA2 along with coeluted variants in the blood samples of patients suffering from beta-thalassemia, indicating differential interactions of the Hb variants with spectrin. PMID- 12737942 TI - The histone deacetylase inhibitor, trichostatin A, reactivates the developmentally silenced gamma globin expression in somatic cell hybrids and induces gamma gene expression in adult BFUe cultures. AB - Somatic cell hybrids that have undergone globin gene switching and developmental silencing of gamma globin expression were treated with the histone deacetylase inhibitor trichostatin A (TSA). Culture of the post-switch hybrids in the presence of TSA reactivated gamma globin expression and concommitantly downregulated beta globin expression, as determined by both mRNA quantitation and immunofluorescent quantitation of gamma globin expressing cells. In contrast, similar treatment of pre-switch hybrids, which were expressing predominantly gamma globin and only small levels of beta globin, had no effect on the relative gamma or beta globin gene expression. In addition, trichostatin A induced gamma gene expression in adult BFUe cultures in a maturation-independent fashion. The results provide direct evidence that inhibition of HDAC activity can alter expression from the human beta globin locus in the adult stage of development. PMID- 12737943 TI - Combined glucose-6-phosphate dehydrogenase and glucosephosphate isomerase deficiency can alter clinical outcome. AB - Glucosephosphate isomerase (GPI) deficiency in humans is an autosomal recessive disorder, which results in nonspherocytic hemolytic anemia of variable clinical expression. A 4-year-old female with severe congenital hemolytic anemia had low red cell GPI activity of 15.5 IU/g Hb (50% of normal mean) indicating GPI deficiency. Subsequent DNA sequence analysis revealed a novel homozygous 921C to G mutation in the GPI gene sequence, predicting a Phe307 to Leu replacement. Strikingly, the red cell GPI activity in this patient was higher than that found in a second patient expressing the same GPI variant, with a more severe clinical phenotype. We propose that the hemolysis in the first patient may be modified by an accompanying deficiency of glucose-6-phosphate dehydrogenase (G6PD). The proband's red cell G6PD activity was reduced at 4.5 IU/g Hb (50% of normal mean) and molecular studies revealed heterozygosity for the G6PD Viangchan mutation and a skewed pattern of X-chromosome inactivation, producing almost exclusive expression of the mutated allele. The G6PD Viangchan variant is characterised by severe enzyme deficiency, but not chronic hemolysis. This study suggests that the metabolic consequences of a combined deficiency of GPI and G6PD might be responsible for a different clinical outcome than predicted for either defect in isolation. PMID- 12737944 TI - Molecular defects in type 3 von Willebrand disease: updated results from 40 multiethnic patients. AB - Type 3 von Willebrand disease (VWD) is characterized by unmeasurable von Willebrand factor (VWF) levels in plasma and platelets and severe hemorrhagic symptoms. We have characterized at the molecular level a group of 40 patients (12 Italians, 14 Iranians, and 14 Indians) to evaluate genetic heterogeneity among these populations. Some of these patients have been previously investigated by us (mutations shown in italics); they are included in this study to provide a more comprehensive pattern of gene defects in type 3 VWD. Patients' DNA were first tested for more frequently reported mutations, then screened by single-strand conformation polymorphism and direct sequence analysis. Fifty gene defects were identified, of which 45 are novel. As expected most of these defects caused null alleles, 17 being nonsense mutations (Q218X, W222X, R365X, R373X, Y610X, W642X, E644X, Q706X, Q1311X, S1338X, Q1346X, Y1542X, R1659X, E1981X, E2129X, R2434X, and Q2544X), 12 small deletions (191delG, 276delT, 788del24, 2016del4, 2157delA, 2269delCT, 2435delC, 4092delAC, 6182delT, 7294delGT, 7683delT, and 8241del9), 4 small insertions (4414insC, 7130insC, 7137insT, and 7674insC), 8 possible splice site mutations (1110(-1)G-->A, 1946(-4)C-->T, 3108(+5)G-->A, 3379(+1)G-->A, 5053(+1)G-->A, 5170(+10)C-->T, 6977(-1)G-->C, and 7729(+7)C-->T), 8 candidate missense mutations (D47H, S85P, D141N, D141Y, C275S, C1071F, C2174G, and C2804Y), and 1 large gene deletion (exons 23-52). Only 2 of these patients have developed alloantibodies against VWF. This study extend our previous finding that mutations responsible for type 3 VWD are scattered throughout the entire VWF gene and that there is no founder effect in these three populations studied. PMID- 12737945 TI - Recombinant murine-activated protein C is neuroprotective in a murine ischemic stroke model. AB - Recombinant mouse protein C was cloned, expressed, purified, and activated by Protac or thrombin. The anticoagulant activities of mouse and human activated protein C (APC) were compared using mouse and human plasma and the neuroprotective properties of murine APC were studied in an ischemic stroke model. Both human APC and mouse APC prolonged the activated partial thromboplastin time in a dose-dependent manner, but mouse APC was sixfold more effective than human APC as an anticoagulant in mouse plasma. Human protein S enhanced prolongation of the APTT clotting time of human plasma by human APC, but not by mouse APC. Hydrolysis of the S-2366 chromogenic substrate by murine APC was essentially identical to human APC. Mouse plasma contains 75 nM protein C. In a murine ischemic stroke model based on middle cerebral artery occlusion, murine APC was highly neuroprotective. The results show that recombinant murine APC is functionally similar to human APC both in vitro and in vivo and that it displays significant species specificity. The results imply that murine APC is notably superior to human APC for studies of murine disease models, including thrombosis and ischemic brain injury. PMID- 12737946 TI - Early gene activation in chronic leukemic B lymphocytes induced toward a plasma cell phenotype. AB - Chronic lymphocytic leukemia (CLL) is characterized by the accumulation of lymphocytes that are arrested at an intermediate stage of B lymphocyte development. CLL B lymphocytes transform (mature) to a plasmacytic phenotype with loss of CD19 and CD20 and the appearance of cytoplasmic immunoglobulin when treated in vitro with phorbol esters. We have used array hybridization technology to describe gene expression patterns for untreated and tetradecanoyl phorbol acetate (TPA)-treated CLL B cells at 5, 10, and 20 min following initial TPA exposure. Three genes, early growth response factor 1 (EGR-1), dual specificity phosphatase 2, and CD69 (early T-cell activation antigen), showed a 2.0-fold or greater increase in mRNA transcription at four or more of six time points in two studies. Upregulation of expression of these genes was confirmed by real-time polymerase chain reaction in the TPA-treated cells of four CLL patients. A progressive increase in gene expression was observed during the 20-min time course for all three genes. In addition, protein expression of EGR-1 and CD69 was increased as measured by immunofluorescence cell analysis. Several genes (PKC, n myc, jun D, and BCL-2) previously reported as overexpressed in CLL lymphocytes were overexpressed in these studies also, but were not altered by TPA treatment. Genes for proteins whose upregulation requires hours of TPA exposure (the 4F2hc component of the L-system amino acid transporter, prohibition, and hsp60) were assessed, and their later expression contrasted with the early expression of EGR 1, dual specificity phosphatase 2, and CD69. EGR-1 encodes a zinc-finger transcription factor that is induced by pokeweed mitogen and TPA and promotes B lymphocyte maturation. The dual specificity phosphatase 2 encodes an enzyme that reverses mitogen activated protein kinase cell activation by dephosphorylation. The CD69 protein is induced by TPA in thymocytes and is a type II transmembrane signaling molecule in hematopoietic cells. These findings suggest that the products of these three genes may be central to early steps in the TPA-induced evolution of CLL B cells to a plasmacytic phenotype. PMID- 12737947 TI - The orchestration of body iron intake: how and where do enterocytes receive their cues? AB - Our understanding of how iron transverses the intestinal epithelium has improved greatly in recent years, although the mechanism by which body iron demands regulate this process remains poorly understood. By critically examining the earlier literature in this field and considering it in combination with recent advances we have formulated a model explaining how iron absorption could be regulated by body iron requirements. In particular, this analysis suggests that signals to alter absorption exert a direct effect on mature enterocytes rather than influencing the intestinal crypt cells. We propose that the liver plays a central role in the maintenance of iron homeostasis by regulating the expression of hepcidin in response to changes in the ratio of diferric transferrin in the circulation to the level of transferrin receptor 1. Such changes are detected by transferrin receptor 2 and the HFE/transferrin receptor 1 complex. Circulating hepcidin then directly influences the expression of Ireg1 in the mature villus enterocytes of the duodenum, thereby regulating iron absorption in response to body iron requirements. In this manner, the body can rapidly and appropriately respond to changes in iron demands by adjusting the release of iron from the duodenal enterocytes and, possibly, the macrophages of the reticuloendothelial system. This model can explain the regulation of iron absorption under normal conditions and also the inappropriate absorption seen in pathological states such as hemochromatosis and thalassemia. PMID- 12737948 TI - A "null allele" mutation is responsible for erythropoietic protoporphyria in an Israeli patient who underwent liver transplantation: relationships among biochemical, clinical, and genetic parameters. AB - Mutations in the human ferrochelatase gene (FECH) are the primary cause of the inborn disorder erythropoietic protoporphyria (EPP). While the majority of the EPP patients exhibit only photosensitivity, a small percentage of patients (approximately 2%) develop liver complications in addition to the cutaneous symptoms. In this study, the FECH gene of an Israeli EPP patient who suffered from EPP-related liver complications was sequenced. A splicing defect IVS10+1, g- >t, which is known to cause the deletion of exon 10, was identified in the index patient as well as in his symptomatic older sister and his asymptomatic mother. Like the other 12 known FECH mutations associated with liver complications, IVS10+1, g-->t is a "null-allele" mutation. Although the two siblings with overt EPP share an identical genotype with respect to both the mutation on one FECH allele and three intragenic single nucleotide polymorphisms, -251G, IVS1-23T, and IVS3-48C on the other allele, the sister of the index patient has so far shown no signs of liver involvement, suggesting that additional factors might account for the liver disease in EPP. PMID- 12737949 TI - Hemochromatosis gene (HFE) mutations in South East Asia: a potential for iron overload. AB - Hereditary hemochromatosis (HH) is an autosomal recessive disease caused by mutations in the HFE gene that mainly affects populations of European descent. Recently a novel mutation (IVS5+1 G-->A) has been described in a Vietnamese patient with HH that was not detected in a European control population. We have developed a novel method to screen for this mutation based on restriction enzyme digestion of a PCR product using a modified forward primer. We have screened 314 Vietnamese people from several ethnic groups and 154 people from Thailand for this mutation and have detected two heterozygotes in the Vietnamese subjects (allele frequency 0.003). Analysis of these heterozygotes indicates that the mutation is on the same haplotype as that found in the original proband. Screening for the widely distributed HFE mutation, H63D, gave an allele frequency of 0.049 in the Vietnamese subjects and 0.032 in the subjects from Thailand. This is the first report of H63D allele frequencies in these populations. We suggest that the presence of the IVS5+1 G-->A and H63D mutations should be considered when investigating iron overload in Vietnamese patients and those of mixed origin as co-inheritance of both mutations is likely to be a risk factor for iron overload. PMID- 12737950 TI - Genetic and transcriptional analysis of spindle checkpoint genes in bone marrow failure patients. AB - The evolution of bone marrow failure syndromes such as aplastic anemia (AA) to clonal hematologic diseases such as myelodysplastic syndrome is well recognized. Cytogenetic abnormalities are commonly seen late events, particularly aneuploidy of chromosomes 7 and 8. A proportion of bone marrow failure patients may also develop aneuploidy that is detectable by fluorescence in situ hybridization but not by standard cytogenetic analysis. The molecular basis for aneuploidy in this setting is currently unknown but may include abnormalities in the mitotic spindle checkpoint. For this reason, we searched for mutations in the mitotic spindle checkpoint genes hBUB1 and hMAD2, and also examined the expression of hBUB1 in cells of bone marrow failure patients. No pathogenic mutations were found in 59 patients. Of 170 bone marrow failure patients, less than one-third expressed hBUB1 transcript. Gene expression profiling confirmed a significant down regulation of hBUB1 message in patients. We conclude that mutations in mitotic spindle checkpoint genes do not account for aneuploidy in marrow failure states. However, we cannot exclude epigenetic inactivation of hBUB1 as a potential mechanism in some patients. PMID- 12737951 TI - Do the designated drivers of college students stay sober? AB - PROBLEM: By numerous accounts, alcohol abuse is considered the number one drug problem facing young people today. Alcohol consumption and its negative consequences, especially those due to drinking and driving, continue to have devastating effects on the college student population. METHOD: This field study examined the blood alcohol concentration (BAC) levels of male and female designated drivers (DD), non-DD, and their respective passengers as they were leaving drinking establishments in a university town. Also investigated were the effects of group size and gender on DD use. RESULTS: A 2 Gender x 2 Driver type (DD vs. non-DD) analysis of variance (ANOVA) for BAC indicated significant main effects for Gender and Driver type, with higher BAC for men and non-DD (p's<.001). A significant Gender x Driver type interaction (p<.05) was primarily due to female DD having lower BAC than male DD. In addition, larger groups were more likely to have a DD. IMPACT ON INDUSTRY: Results indicate that the success of DD programs may be influenced by group size and a DD's gender. While larger groups are more likely to have a DD, students riding home with a male DD may still be at risk for the negative consequences of drunk driving. PMID- 12737952 TI - Is there a link between children's motor abilities and unintentional injuries? AB - PROBLEM: The common view is that clumsy children experience unintentional injury more frequently. Empirical evidence supporting this position is mixed. METHOD: One hundred 6- and 8-year-olds completed a battery of nine tasks designed to assess motor ability. Mothers completed a lifetime injury history measure about their children and families completed a 2-week injury diary assessing frequency and severity of daily injuries. RESULTS: Internal reliability for the motor ability battery was good. Correlations between motor ability measures and injury risk were nonsignificant and near zero. DISCUSSION: Motor ability does not appear to be directly related to injury risk. Possible explanations include: (a) coordinated and clumsy children engage in hazardous activities with differing frequency; or (b) other individual difference factors may interact with motor ability to explain children's injury risk. IMPACT ON INDUSTRY: Children's motor abilities do not appear to be directly linked to rate of unintentional injury, but instead may influence risk for injury in conjunction with other factors. Results could have implications to the engineering of children's toys and playground equipment and to the design of appropriate supervision strategies for children engaging in potentially dangerous activities. PMID- 12737953 TI - Mental models of safety: do managers and employees see eye to eye? AB - PROBLEM: Disagreements between managers and employees about the causes of accidents and unsafe work behaviors can lead to serious workplace conflicts and distract organizations from the important work of establishing positive safety climate and reducing the incidence of accidents. METHOD AND RESULTS: In this study, the authors examine a model for predicting safe work behaviors and establish the model's consistency across managers and employees in a steel plant setting. Using the model previously described by Brown, Willis, and Prussia (2000), the authors found that when variables influencing safety are considered within a framework of safe work behaviors, managers and employees share a similar mental model. The study then contrasts employees' and managers' specific attributional perceptions. Findings from these more fine-grained analyses suggest the two groups differ in several respects about individual constructs. Most notable were contrasts in attributions based on their perceptions of safety climate. When perceived climate is poor, managers believe employees are responsible and employees believe managers are responsible for workplace safety. However, as perceived safety climate improves, managers and employees converge in their perceptions of who is responsible for safety. IMPACT ON INDUSTRY: It can be concluded from this study that in a highly interdependent work environment, such as a steel mill, where high system reliability is essential and members possess substantial experience working together, managers and employees will share general mental models about the factors that contribute to unsafe behaviors, and, ultimately, to workplace accidents. It is possible that organizations not as tightly coupled as steel mills can use such organizations as benchmarks, seeking ways to create a shared understanding of factors that contribute to a safe work environment. Part of this improvement effort should focus on advancing organizational safety climate. As climate improves, managers and employees are likely to agree more about the causes of safe/unsafe behaviors and workplace accidents, ultimately increasing their ability to work in unison to prevent accidents and to respond appropriately when they do occur. Finally, the survey items included in this study may be useful to organizations wishing to conduct self-assessments. PMID- 12737954 TI - A survey-based system for safety measurement and improvement. AB - PROBLEM: A task force sought to develop a method for safety measurement that is reliable and valid and provides a framework for improvement efforts. METHOD: Over a 10-year period, through working in a chemical company with about 6,000 employees and over 50 plants, the authors researched the use of employee surveys to measure safety and as a diagnostic tool for improvement efforts. RESULTS: The statistical studies indicate that this survey, which evolved from the Minnesota Safety Perception Survey, is both reliable and valid as a measurement tool. The survey measures important components of the management system including (a) management's demonstration of commitment to safety, (b) education and knowledge of the workforce, (c) effectiveness of the supervisory process, and (d) employee involvement and commitment. This study also describes anecdotal evidence that the diagnostic element of the survey enables the development of effective action plans to improve safety performance. This evidence includes ratings of the process by plant managers who have used it. IMPACT ON INDUSTRY: The survey and related methods have helped to improve safety performance in several companies. PMID- 12737955 TI - Technology use and psychosocial factors in the self-reporting of musculoskeletal disorder symptoms in call center workers. AB - PROBLEM: Evidence exists to suggest that working with computer terminals and keyboards is associated with the development and exacerbation of a range of musculoskeletal disorders (MSDs). Research has suggested that psychosocial factors may be of more importance than computer use factors in the development of MSDs in the computing workplace. METHOD: A questionnaire-based study was undertaken to test this hypothesis in a sample of 67 call center workers. A Spearman's rho was performed on the data, as well as chi(2) tests on the individual variables. RESULTS: It was found that overall, computer use factors were significantly associated with self-reporting of MSD symptoms, whereas psychosocial factors had no such association. However, certain individual psychosocial factors emerged as having a significant association with MSD symptoms. DISCUSSION: The findings suggest that specific areas of call center work have associations with poor worker health and possible long-term MSD problems. These areas appear to be workload and particular management-worker relations, rather than computer use. SUMMARY: This suggests that the relationship among computer use, psychosocial factors, and MSD is a complex area that would benefit from more research. IMPACT ON INDUSTRY: This study raises issues about the importance of psychosocial and organizational factors and employee welfare in the call center working environment. PMID- 12737956 TI - Preventing collisions involving surface mining equipment: a GPS-based approach. AB - PROBLEM: An average of three workers a year are killed in surface mining operations when a piece of haulage equipment collides with another smaller vehicle or a worker on foot. Another three workers are killed each year when haulage equipment backs over the edge of a dump point or stockpile. Devices to monitor the blind areas of mining equipment are needed to provide a warning to operators when a vehicle, person, or change in terrain is near the equipment. METHOD: A proximity warning system (PWS) based on the global positioning system (GPS) and peer-to-peer communication has been developed to prevent collisions between mining equipment, small vehicles, and stationary structures. RESULTS: A final system was demonstrated using one off-highway haul truck, three smaller vehicles, and various stationary structures at a surface mining operation. The system successfully displayed the location of nearby vehicles and stationary structures and provided visual and audible warnings to the equipment operator when they were within a preset distance. SUMMARY: Many surface mining operations already use GPS technology on their mobile equipment for tracking and dispatch. Our tests have shown that it is feasible to add proximity warning to these existing systems as a safety feature. Larger scale and long-term tests are needed to prove the technology adequately. IMPACT ON INDUSTRY: A PWSs that incorporates a combination of technologies could significantly reduce accidents that involve collisions or driving over an edge at surface mining operations. PMID- 12737957 TI - Risk-mitigating beliefs, risk estimates, and self-reported speeding in a sample of Australian drivers. AB - PROBLEM: Research suggests that people who engage in risk-taking behaviors often hold specific beliefs that can mitigate or reduce their perceptions of risk associated with those behaviors. METHOD: A scale was developed (Speeding Risk Belief Scale (SRBS)) to assess beliefs about speeding-related risk and predict self-reported speeding in a random-digit telephone survey of 800 South Australian drivers between the ages of 16 and 50. RESULTS: The scale was internally consistent, and path analyses showed it to be associated with self-reported speeding, both directly and indirectly through participants' estimates of speeding-related risk. DISCUSSION: Origins of risk-mitigating beliefs and the extent to which they may be causally linked with speeding are discussed, and recommendations are made for future research. IMPACT ON INDUSTRY: This research has strong implications for the conduct of countermeasure campaigns that disseminate information on speeding-related risk. PMID- 12737958 TI - Management practices as antecedents of safety culture within the trucking industry: similarities and differences by hierarchical level. AB - PROBLEM: A homogeneous perception of safety is important for the achievement of a strong safety culture; however, employees may differ in their safety perceptions, depending on their position and/or hierarchical level within the organization. Moreover, there is limited information on the antecedents of safety culture. This study examines how safety training, driver scheduling autonomy, opportunity for safety input, and management commitment to safety influence individuals' perceptions of safety culture. METHOD: Data for this study were drawn from 116 trucking firms, stratified by three safety performance levels. The data were collected from drivers (lowest hierarchical level), dispatchers (medium hierarchical level), and safety directors (highest hierarchical level), regarding their perceptions of their respective corporate safety cultures. Perceptions of safety culture were analyzed through a linear regression using dummy variables to differentiate among the three hierarchical groups. The resulting model allowed for examination of the specific antecedents of safety culture for the three employee groups and the extent to which the hierarchical groups were in agreement with each other. RESULTS: Driver fatigue training, driver opportunity for safety input, and top management commitment to safety were perceived to be integral determinants of safety culture in all three groups. IMPACT ON INDUSTRY: Trucking firms seeking to strengthen employees' perceptions of safety culture might begin by improving these safety management practices while appreciating that they may have a different impact depending on the employee's hierarchical position (e.g., drivers' perceptions of safety culture are more influenced by top management commitment and driver fatigue training). A fourth safety practice examined, driver scheduling autonomy, was not found to be instrumental in shaping safety culture for any of the three hierarchical levels. Consistent with previous research, implementation of stronger safety cultures should result in fewer accidents. PMID- 12737959 TI - Age differences in safety attitudes and safety performance in Hong Kong construction workers. AB - PROBLEM: Safety in the construction industry is a major issue in Hong Kong, representing about 46% of all occupational injuries in 1998. This study explored linear and curvilinear relations between age and safety performance (accident rates and occupational injuries), as well as safety attitudes, in construction workers in Hong Kong. METHOD: A Chinese version of the Safety Attitude Questionnaire (SAQ by Donald & Canter) was developed and administered to a sample of Chinese construction workers (N=374, 366 males, 8 females) from 27 construction sites. RESULTS: Accident rates were not related to age. Occupational injuries were related to age in a curvilinear manner, with injuries at first increasing with age, then decreasing. Two safety attitude scales were related to age with older workers exhibiting more positive attitudes to safety. If age and tenure are controlled, some attitude scales are predictors of safety performance. IMPACT ON INDUSTRY: Management/supervisors, team leaders, and workers are all responsible for safety, and any negative bias toward older construction workers is unfounded. PMID- 12737960 TI - An empirical investigation of European drivers' self-assessment. AB - PROBLEM: Evaluating motorists through self-assessment has attracted much interest in recent literature, which is mainly due to the profound impact various parameters of self-assessment can have on the way motorists deal with hazardous traffic situations. Much of the previous work in this area has been hampered both by the lack of adequate sample sizes and, because of the small samples, the evaluation methodologies used. METHOD: This paper extends previous research in two significant directions: (a) it uses the SARTRE 2 database, which provides more than 17,000 questionnaires from most European countries; and (b) it employs the ordered probit modeling approach, which recognizes the latent nature of self assessment and explicitly links its dimensions to a set of relevant explanatory variables such as age, gender, region, and income. RESULTS: The results indicate that drivers who rate themselves as both more dangerous and faster than others are, generally, younger men, with higher incomes, break the speed limit more frequently, avoid wearing seat belts, and have been involved in more accidents in the past than other drivers. Interestingly, more experienced and more highly educated drivers assess their driving as less dangerous, but admit to driving faster than other drivers. IMPACT ON INDUSTRY: The methodology used and the results obtained can be a significant help in identifying drivers with high and low self-assessment ratings, which can be useful in planning and implementing road safety information campaigns. PMID- 12737961 TI - The "Goldilocks model" of overtime in construction: not too much, not too little, but just right. AB - PROBLEM: Little research exists on the relationship between working overtime and possible adverse health and safety outcomes for construction workers. METHOD: Five focus-group discussions were conducted with construction workers from around the United States. From the analyzed transcripts, a model of overtime was developed. RESULTS: The model includes three dominant themes: (1) work organization issues [(a) definitions of overtime, (b) scheduling, and (c) economic conditions], (2) why workers choose to work overtime [(a) management expectations, (b) career, and (c) money], and (3) the effects of working overtime [(a) health and safety, including sleep deprivation, injury, fatigue, and stress, and (b) productivity]. DISCUSSION: Health and safety is only one of the adverse outcomes related to working too much overtime. A list of worker-inspired recommendations for addressing overtime issues is provided. IMPACT ON THE INDUSTRY: Both employers and workers need to better understand the potential adverse effects of working too much overtime. PMID- 12737962 TI - Potential of scrap tire rubber as lightweight aggregate in flowable fill. AB - Flowable fill is a self-leveling and self-compacting material that is rapidly gaining acceptance and application in construction, particularly in transportation and utility earthworks. When mixed with concrete sand, standard flowable fill produces a mass density ranging from 1.8 to 2.3 g/cm(3) (115-145 pcf). Scrap tires can be granulated to produce crumb rubber, which has a granular texture and ranges in size from very fine powder to coarse sand-sized particles. Due to its low specific gravity, crumb rubber can be considered a lightweight aggregate. This paper describes an experimental study on replacing sand with crumb rubber in flowable fill to produce a lightweight material. To assess the technical feasibility of using crumb rubber, the fluid- and hardened-state properties of nine flowable fill mixtures were measured. Mixture proportions were varied to investigate the effects of water-to-cement ratio and crumb rubber content on fill properties. Experimental results indicate that crumb rubber can be successfully used to produce a lightweight flowable fill (1.2-1.6 g/cm(3) [73 98 pcf]) with excavatable 28-day compressive strengths ranging from 269 to 1194 kPa (39-173 psi). Using a lightweight fill reduces the applied stress on underlying soils, thereby reducing the potential for bearing capacity failure and minimizing soil settlement. Based on these results, a crumb rubber-based flowable fill can be used in a substantial number of construction applications, such as bridge abutment fills, trench fills, and foundation support fills. PMID- 12737963 TI - Mechanochemical treatment to recycling asbestos-containing waste. AB - Numerous industrial and experimental facilities have been set up, particularly in the last ten years, as a result of studies and researches on treating asbestos containing waste (ACW) to stabilise it and to enable its reuse. Some of the stabilisation processes reduce the hazards of ACW by imprisoning in a cement or resonoid matrix. Other processes modify the fibrous structure of asbestos and transform it into an inert substance. One such inactivation process is mechanochemical transformation. This new technology is extremely interesting both economically and industrially, especially in view of the European Directive 1999/3/CE of 24/4/99, which provides for the obligatory treatment of all types of waste material before its disposal. PMID- 12737964 TI - Multi-regime transport model for leaching behavior of heterogeneous porous materials. AB - Utilization of secondary materials in civil engineering applications (e.g. as substitutes for natural aggregates or binder constituents) requires assessment of the physical and environment properties of the product. Environmental assessment often necessitates evaluation of the potential for constituent release through leaching. Currently most leaching models used to estimate long-term field performance assume that the species of concern is uniformly dispersed in a homogeneous porous material. However, waste materials are often comprised of distinct components such as coarse or fine aggregates in a cement concrete or waste encapsulated in a stabilized matrix. The specific objectives of the research presented here were to (1) develop a one-dimensional, multi-regime transport model (i.e. MRT model) to describe the release of species from heterogeneous porous materials and, (2) evaluate simple limit cases using the model for species when release is not dependent on pH. Two different idealized model systems were considered: (1) a porous material contaminated with the species of interest and containing inert aggregates and, (2) a porous material containing the contaminant of interest only in the aggregates. The effect of three factors on constituent release were examined: (1) volume fraction of material occupied by the aggregates compared to a homogeneous porous material, (2) aggregate size and, (3) differences in mass transfer rates between the binder and the aggregates. Simulation results confirmed that assuming homogeneous materials to evaluate the release of contaminants from porous waste materials may result in erroneous long-term field performance assessment. PMID- 12737965 TI - Monitoring the stabilization of municipal solid waste incineration fly ash by phosphation: mineralogical and balance approach. AB - The application of a micro-characterization protocol coupled with a balance approach has allowed the relevant monitoring of a phosphation process for fly ash produced by municipal solid waste incineration. The three main steps of this process consist in removing the salts (chlorides, sulfates) by dissolution at basic pH, phosphation of the residue to trap metals, and its calcination to destroy dioxin-like compounds. The chemical and mineralogical balances compiled on the samples after each step of the process validate these main objectives and highlight the wide phosphorus distribution throughout the sample during the phosphation process, as well as the formation of apatite-type crystallized phosphates. During calcination, the increase in the proportion of crystallized phosphates apatite and whitlockite is largely attributable to the presence of an available calcium source, corresponding to the calcite formed during washing. The metals Pb and Zn, initially distributed in the silicate and carbonate phases, are broadly redistributed in the phosphate neoformations after carbonate dissolution, thus guaranteeing a more permanent stabilization. PMID- 12737966 TI - Recovery of valuable metals from electronic and galvanic industrial wastes by leaching and electrowinning. AB - In the present paper, a study on laboratory scale to perform a treatment for valuable metals recovery from electronic and galvanic industrial wastes, is reported. The characterisation of the waste, performed by XRD, SEM, EDX and chemical analysis, showed a high metals content in the sludge, such as Cu, Ni, Mn, Pb, Sn, W. A leaching process, coupled by electrowinning, is then proposed in order to reduce the volume of the waste material and to recover selectively valuable metals, such as Cu and Ni. During the leaching step, carried out by using H(2)SO(4), several factors were investigated (acid concentration, temperature and time of treatment). The leached liquor has been successfully treated with an electrowinning process, to recover copper and nickel. The copper and nickel depositions, were performed in acid and alkaline conditions, respectively. The Faraday yield was of about 95%. The energy consumption was 2.13 and 4.43 kWh per kg of copper and nickel recovered, respectively. At the end of the process, about 94-99% of the initial content of Cu and Ni was recovered at the cathode. The experimental results obtained, showed the technical feasibility of the process. PMID- 12737967 TI - Destruction of PAHS from soil by using pressurized hot water extraction coupled with supercritical water oxidation. AB - Chemical processes utilizing water both as extraction solvent and reaction medium are promising "Green Chemistry" alternatives to conventional techniques. Equipment for on-line coupled hot water extraction and supercritical water oxidation was constructed to extract polyaromatic hydrocarbons and toluene from sea sand followed by oxidation using hydrogen peroxide. The effectiveness of the technique is based on the physico-chemical properties of heated and pressurized water. Extraction efficiency increased with temperature and time; the best results were obtained at 300 degrees C with 40 min extraction time. In the oxidation stage, conversion of the PAHs increased with reaction time and oxidant concentration and the best conversion (97.0-99.9%, depending on the compound) was obtained at 425 degrees C with 43 s reaction time. Benzaldehyde and benzoic acid were the most abundant reaction intermediates in the oxidation process. In addition, phenol, p-cresol, and benzyl alcohol were found as intermediates. The intermediates originated mainly from toluene, which was present in much greater concentration than PAHs in the reaction medium. PMID- 12737968 TI - Air stripping of ammonia from pig slurry: characterisation and feasibility as a pre- or post-treatment to mesophilic anaerobic digestion. AB - The objective of the present paper has been to study the effect of pig slurry waste type, fresh or anaerobically digested, and the effect of initial pH on ammonia air stripping from pig slurry waste at high temperature (80 degrees C). Stripping process as pre- or post-treatment to anaerobic digestion has been also evaluated. Treatment performances differ according to pig slurry type. When fresh pig slurry is used, despite working at 80 degrees C, a high initial pH (11.5) is required for complete ammonia removal. On the other hand, for digested pig slurry, complete ammonia removal without pH modification is possible and organic matter significantly less contaminates recovered ammonia salt. Batch anaerobic tests showed that ammonia air stripping is not an advisable pre-treatment to pig slurry anaerobic digestion. PMID- 12737969 TI - Gaseous emissions from ceramics manufactured with urban sewage sludge during firing processes. AB - The re-use of sewage sludge without any treatment as primary material-mixed with clays-in order to obtain structural ceramics for buildings has been successfully improved. In the Ecobrick project, the firing of a mixture of specific percentages of three components (clays, sludges and forest debris) resulted in a lighter and more thermal and acoustic insulating brick, compared with conventional clay-bricks. Volatile organic compounds (VOC) emission from the manufacturing of ceramics is the most important aspect to control. In the Ecobrick project VOC emissions were monitored by using a bench-scale furnace. The study was conducted using an EPA recommended sampling train and portable sampling tubes that were thermally desorbed and analyzed by gas chromatography/mass spectrometry. Drying of raw sewage-sludge and firing processes were considered separately. In this paper, we present VOC emissions coming from the firing step of the Ecobrick production. PMID- 12737973 TI - Global phytochemistry: the Egyptian experience. PMID- 12737970 TI - Determination of Cu and Ni incorporation ratios in Portland cement clinker. AB - Cu and Ni are metals found in galvanic sludges; these sludges are considered hazardous due to their heavy metal content. The main objective of this work is to determine the incorporation amount of Cu and Ni in Portland clinker when a galvanic sludge containing these metals is added to the clinker raw-material. The influence of this addition on the clinkering reactions is evaluated as well as the possibility of co-incinerating galvanic sludges containing Cu and Ni in rotary cement kilns. This study also characterizes the galvanic sludge. Samples were prepared by additions from 0.25 to 5 wt.% of a galvanic sludge to an industrial clinker raw-material. The clinkering process was simulated in a laboratory device. The following techniques were applied to characterize the raw materials and the products of the tests: chemical analysis, differential thermal analysis (DTA) and thermogravimetric analysis (TG). Leaching tests were performed in the produced clinker samples in order to verify the incorporation of the studied metals in the clinker structure. The results led to the conclusion that additions of up to 2 wt.% of a galvanic sludge containing 2.4 wt.%Cu and 1.2 wt.% Ni to clinker raw-material do not affect the clinkering reactions and that these metals are totally incorporated into the clinker. PMID- 12737974 TI - Pectate lyase activity during ripening of banana fruit. AB - Pectate lyase (PEL) activity was demonstrated in ripe banana fruits on supplementing the homogenizing medium with cysteine and Triton X-100. The enzyme was characterized on the basis of alkaline pH optimum, elimination of the activity by EDTA and activation by Ca(2+). PEL activity was not detected in preclimacteric banana fruits. PEL activity increased progressively from early climacteric and reached maximum level at climacteric peak and declined in post climacteric and over ripened fruits. Replacing pectate with pectin in PEL assay manifested enzyme activity even in preclimacteric fruits. In contrast to PEL, polygalacturonase activity progressively increased during fruit ripening even in postclimacteric fruits. PMID- 12737975 TI - Cys/Gly-rich proteins with a putative single chitin-binding domain from oat (Avena sativa) seeds. AB - Through a reliable and repeatable procedure based on solid-phase extraction techniques, a protein fraction (P fraction) rich in Cys/Gly residues was extracted and captured from oat (Avena sativa L.) seeds. Quantitative amino acid analysis and MS of the P fraction indicated that it contains a series of heterogeneous Cys/Gly-rich proteins with molecular masses of 3.6-4.0 kDa. Preliminary results from bioassays showed that these proteins possess weak to moderate antifungal properties to some fungal strains. From this fraction, a new polypeptide, designated avesin A, was purified and sequenced by Edman degradation. Avesin A consists of 37 amino-acid residues, with 10 glycine residues and eight cysteine residues forming disulfide bridges, and contains a single chitin-binding domain, which indicates that avesin A is a new member of the putative chitin-binding proteins. Avesin A is the first identified hevein like small protein from cereal grains. PMID- 12737976 TI - An esterase is involved in geraniol production during palmarosa inflorescence development. AB - Total incorporation of exogenously administered [2-14C]acetate into essential oil of palmarosa (Cymbopogon martinii) was found to be relatively higher than that of either [U-14C]sucrose or [U-14C]glucose during inflorescence development. Among the major essential oil constituents, biogenesis of geranyl acetate was much higher than that of geraniol. Alkaline hydrolysis of [14C]labeled geranyl acetate revealed that the majority of the label incorporated into geranyl acetate was present in the geraniol moiety, indicating that only newly synthesized geraniol gets acetylated to form geranyl acetate. Geranyl acetate cleaving esterase (GAE) activity followed a similar pattern during both in vivo and in vitro inflorescence development, with maximum activity at immature inflorescence stages, suggesting the involvement of GAE in geraniol production during inflorescence development. Five esterase isozymes (Est-A to E) were detected in the enzymic fraction of palmarosa inflorescence and all showed GAE activity, with Est-B being significantly increased during inflorescence development. The role of GAE in geraniol production and improving the palmarosa oil quality is discussed. PMID- 12737977 TI - Fragrance chemistry, nocturnal rhythms and pollination "syndromes" in Nicotiana. AB - GC-MS analyses of nocturnal and diurnal floral volatiles from nine tobacco species (Nicotiana; Solanaceae) resulted in the identification of 125 volatiles, including mono- and sesquiterpenoids, benzenoid and aliphatic alcohols, aldehydes and esters. Fragrance chemistry was species-specific during nocturnal emissions, whereas odors emitted diurnally were less distinct. All species emitted greater amounts of fragrance at night, regardless of pollinator affinity. However, these species differed markedly in odor complexity and emission rates, even among close relatives. Species-specific differences in emission rates per flower and per unit fresh or dry flower mass were significantly correlated; fragrance differences between species were not greatly affected by different forms of standardization. Flowers of hawkmoth-pollinated species emitted nitrogenous aldoximes and benzenoid esters on nocturnal rhythms. Four Nicotiana species in section Alatae sensu strictu have flowers that emit large amounts of 1,8 cineole, with smaller amounts of monoterpene hydrocarbons and alpha-terpineol on a nocturnal rhythm. This pattern suggests the activity of a single biosynthetic enzyme (1,8 cineole synthase) with major and minor products; however, several terpene synthase enzymes could contribute to total monoterpene emissions. Our analyses, combined with other studies of tobacco volatiles, suggest that phenotypic fragrance variation in Nicotiana is shaped by pollinator- and herbivore-mediated selection, biosynthetic pathway dynamics and shared evolutionary history. PMID- 12737978 TI - Actinidia arguta: volatile compounds in fruit and flowers. AB - More than 240 compounds were detected when the volatile components of the flowers and the fruit from several Actinidia arguta genotypes were investigated. Around 60-70 different compounds were extracted from individual tissues of each genotype. Two different methods of volatile sampling (headspace and solvent) favoured different classes of compounds, dependent upon their volatilities and solubilities in the flower or fruit matrices. The compounds extracted from flowers largely comprised linalool derivatives including the lilac aldehydes (12a d) and alcohols (13a-d), 2,6-dimethyl-6-hydroxyocta-2,7-dienal (8), 8 hydroxylinalool (9), sesquiterpenes, and benzene compounds that are presumed metabolites of phenylalanine and tyrosine. Extracts of fruit samples contained some monoterpenes, but were dominated by esters such as ethyl butanoate, hexanoate, 2-methylbutanoate and 2-methylpropanoate, and by the aldehydes hexanal and hex-E2-enal. A number of unidentified compounds were also detected, including 8 from flowers that are so closely related that they are either isomers of one compound or two or more closely related compounds. This is the first report of the presence of a range of linalool derivatives in Actinidia. PMID- 12737979 TI - Development of an ELISA approach for the determination of flavodoxin and ferredoxin as markers of iron deficiency in phytoplankton. AB - Quantification of the iron-nutritional status of phytoplankton is of great interest not only for the study of oceans but also for fresh waters. Flavodoxin is a small flavoprotein proposed as a marker for iron deficiency, since it is induced as a consequence of iron deprivation, replacing the iron-sulphur protein ferredoxin. Flavodoxin and ferredoxin have been frequently used as markers for determination of iron deficiency in phytoplankton. Using purified flavodoxin and ferredoxin from Scenedesmus vacuolatus and polyclonal antibodies against both proteins, individual ELISA tests have been developed. The assays have a linear response in the range of 30-600 ng/ml of protein. PMID- 12737980 TI - Regional and habitat differences in 7-methyljuglone content of Finnish Drosera rotundifolia. AB - The concentration of 7-methyljuglone was studied in the round-leaved sundew Drosera rotundifolia L. collected from different regions in Northern Finland. Samples for analysis were collected from peat bogs and sandpit habitats. The mean concentration of 7-methyljuglone varied from 1.0 to 2.3% of dry weight. Variation between years in the amount of 7-methyljuglone was significant in plants growing on sand, and in the northernmost region studied. Overall, the variation in the production of 7-methyljuglone among different populations of round-leaved sundew in Northern Finland was rather low. The variation between years in the production of 7-methyljuglone was more significant. PMID- 12737981 TI - Preparation and biological assessment of hydroxycinnamic acid amides of polyamines. AB - Many plants contain hydroxycinnamic acid conjugates of polyamines that are remarkably similar in general structure to the acylated polyamines found in spider and wasp toxins. In an effort to determine whether these compounds might play a role in the chemical defense of plants against arthropod pests we synthesized a variety of analogues of the coumaric (4-hydroxycinnamic) acid conjugates of di-, tri-, and tetraamines using common protection and acylation strategies. N(1)- and N(8)-coumaroyl spermidine were tested in feeding trials with insect larvae including the European corn borer (Ostrinia nubilalis), the tobacco budworm (Heliothis verescens) and the oblique banded leaf roller (Choristoneura rosaceana). Antifeedant assays with the rice weevil Sitophilus oryzae were also performed. Neither the naturally occurring coumaric acid conjugates of polyamines nor their analogues showed notable toxicity towards insects, despite precautions to maintain these easily oxidized materials in the wet diet. However, more direct bioassays of these compounds on glutamate dependent neuroreceptors including the deep abdominal extensor muscles of crayfish, or mammalian NMDA, delta2, and AMPA receptors, clearly showed that these compounds were inhibitory. N(1)-Coumaoryl spermine, a dodecyl and a cyclohexyl analogue were especially active at NMDA NR1/NR2B receptors. The latter had an IC(50) of 300 microM in the crayfish. N(1)-Coumaroyl spermine had an IC(50) in the crayfish preparation of 70-300 microM and against the mammalian NR1/NR2B receptor of 38 nM. Structure-activity variations show similar trends of length and hydrophobicity as has been seen previously with analogues of spider toxins. We conclude from this work that while the coumaric acid polyamine conjugates are active when directly applied to neuroreceptors, they show no overt toxicity when ingested by insect larvae. PMID- 12737982 TI - Activity-guided isolation of the chemical constituents of Muntingia calabura using a quinone reductase induction assay. AB - Activity-guided fractionation of an EtOAc-soluble extract of the leaves of Muntingia calabura collected in Peru, using an in vitro quinone reductase induction assay with cultured Hepa 1c1c7 (mouse hepatoma) cells, resulted in the isolation of a flavanone with an unsubstituted B-ring, (2R,3R)-7-methoxy-3,5,8 trihydroxyflavanone (5), as well as 24 known compounds, which were mainly flavanones and flavones. The structure including absolute stereochemistry of compound 5 was determined by spectroscopic (HRMS, 1D and 2D NMR, and CD spectra) methods. Of the isolates obtained, in addition to 5, (2S)-5-hydroxy-7 methoxyflavanone, 2',4'-dihydroxychalcone, 4,2',4'-trihydroxychalcone, 7 hydroxyisoflavone and 7,3',4'-trimethoxyisoflavone were found to induce quinone reductase activity. PMID- 12737983 TI - Effects of black-eyed pea trypsin/chymotrypsin inhibitor on proteolytic activity and on development of Anthonomus grandis. AB - The cotton boll weevil Anthonomus grandis (Boheman) is one of the major pests of cotton (Gossypium hirsutum L.) in tropical and sub-tropical areas of the New World. This feeds on cotton floral fruits and buds causing severe crop losses. Digestion in the boll weevil is facilitated by high levels of serine proteinases, which are responsible for the almost all proteolytic activity. Aiming to reduce the proteolytic activity, the inhibitory effects of black-eyed pea trypsin/chymotrypsin inhibitor (BTCI), towards trypsin and chymotrypsin from bovine pancreas and from midguts of A. grandis larvae and adult insects were analyzed. BTCI, purified from Vigna unguiculata (L.) seeds, was highly active against different trypsin-like proteinases studied and moderately active against the digestive chymotrypsin of adult insects. Nevertheless, no inhibitory activity was observed against chymotrypsin from A. grandis larval guts. To test the BTCI efficiency in vivo, neonate larvae were reared on artificial diet containing BTCI at 10, 50 and 100 microM. A reduction of larval weight of up to approximately 54% at the highest BTCI concentration was observed. At this concentration, the insect mortality was 65%. This work constitutes the first observation of a Bowman-Birk type inhibitor active in vitro and in vivo toward the cotton boll weevil A. grandis. The results of bioassays strongly suggest that BTCI may have potential as a transgene protein for use in engineered crop plants modified for heightened resistance to the cotton boll weevil. PMID- 12737984 TI - Structural diversity of fructans from members of the order Asparagales in New Zealand. AB - The water-soluble carbohydrates (WSCs) extracted from the underground parts of Arthropodium cirratum, Astelia banksii, Bulbinella hookeri, Dianella nigra and Xeronema callistemon, and the flower stem of Phormium tenax have been investigated. Extracts of A. cirratum, B. hookeri, D. nigra and P. tenax contained 108.4, 28.1, 41.9 and 29.7 mg gFW(-1) WSCs, respectively. Thin-layer chromatography (TLC) showed that these species contained fructans. Extracts of A. banksii and X. callistemon each contained 11 mg gFW(-1) WSCs or less, and TLC detected only monosaccharides and sucrose. Reverse-phase (RP) HPLC and glycosyl linkage analysis showed that extracts of B. hookeri contained predominantly a linear series of fructans, with 1-linked and terminal fructofuranosyl (Fruf) residues and terminal glucopyranose (Glcp). RP-HPLC of extracts of A. cirratum, D. nigra and P. tenax showed a more complex pattern of oligosaccharides. Linkage analysis showed that these extracts contained fructans with 1-linked Fruf, 6 linked Fruf and 1,6-branched Fruf. Extracts of D. nigra and P. tenax contained 6 Glcp with only trace amounts of terminal Glcp, while A. cirratum contained mostly terminal Glcp with a trace of 6-Glcp. Differences in fructan and fructo oligosaccharide structure between species shown in this study, together with published data, suggests that there are differences in the activities of the various fructosyltransferases responsible for biosynthesis of fructans in species within the Asparagales. PMID- 12737985 TI - Compound-specific deltaD-delta13C analyses of n-alkanes extracted from terrestrial and aquatic plants. AB - Stable hydrogen and carbon isotopic compositions of individual n-alkanes were determined for various terrestrial plants (33 samples including 27 species) and aquatic plants (six species) in natural environments from Japan and Thailand. In C3 plants, n-alkanes extracted from angiosperms have a deltaD value of -152+/-26 per thousand (relative to Standard Mean Ocean Water [SMOW]) and delta13C value of -36.1+/-2.7 per thousand (relative to Peedde Belemnite [PDB]), and those from gymnosperms have a deltaD value of -149+/-16 per thousand and delta13C value of 31.6+/-1.7 per thousand. Angiosperms have n-alkanes depleted in 13C relative to gymnosperms. n-Alkanes from C4 plants have a deltaD value of -171+/-12 per thousand and delta13C value of -20.5+/-2.1 per thousand, being a little depleted in D and much enriched in 13C compared to C3 plants. n-Alkanes of CAM plants are a little depleted in D and vary widely in delta13C relative to those of C3 and C4 plants. In aquatic plants, n-alkanes from freshwater plants have a deltaD value of -187+/-16 per thousand and delta13C value of -25.3+/-1.9 per thousand, and those from seaweeds have a deltaD value of -155+/-34 per thousand and delta13C value of -22.8+/-1.0 per thousand. All n-alkanes from various plant classes are more depleted in D and 13C relative to environmental water and bulk tissue, respectively. In addition, the hydrogen and carbon isotopic fractionations during n-alkane synthesis are distinctive for these various plant classes. While C3 plants have smaller isotopic fractionations in both D and 13C, seaweed has larger isotopic fractionations. PMID- 12737989 TI - Identification of HLA-A*0201-restricted cytotoxic T-cell epitopes of Trypanosoma cruzi TcP2beta protein in HLA-transgenic mice and patients. AB - Trypanosoma cruzi-specific cytotoxic T-lymphocyte (CTL) responses are critical in the control of parasite growth and will play an important part in therapeutic and prophylactic T. cruzi vaccines. The identification of parasite-specific epitopes that are efficiently recognized by CTLs is the first step in the development of future vaccines. HLA-A2 transgenic mice (HHD) were shown to provide a powerful model for studying the induction of HLA-A*0201-restricted immune responses in vivo, since these mice are endowed with a CTL repertoire representative of HLA A2.1 individuals. Here, we describe the immunological characterization of T-cell epitopes of the T. cruzi ribosomal P2 protein (TcP2beta) that are recognized by HLA-A*0201-restricted CTLs in HLA-transgenic mice and humans. Epitopes identified in the present study do not share sequence homology with the homologous human or murine counterparts and so they should not induce any autoreactive response. Moreover, HHD mice vaccinated with these peptide epitopes have reduced parasitemia after challenge with a lethal T. cruzi infection. Hence, these epitopes represent potential subunit components of multi-protein vaccines to prevent Chagas' disease. PMID- 12737990 TI - Leishmania tropica in the black rat (Rattus rattus): persistence and transmission from asymptomatic host to sand fly vector Phlebotomus sergenti. AB - Black rats (Rattus rattus) receiving Leishmania tropica injected intradermally into the ear were studied for the persistence of parasites and infectivity to natural sand fly vector. The mammalian host, the parasite, and the vector all originated from the endemic focus of Urfa, Turkey. Rats did not develop lesions or any apparent signs of disease, although at the site of inoculation they harboured live parasites capable of infecting sand flies. The number of L. tropica amastigotes detected in the inoculated ear by quantitative real-time PCR ranged from 5 x 10(3) to 10(6). Parasite DNA was also present in the tail and contralateral ear, sites distant from inoculation. After feeding on the ears of asymptomatic rats, Phlebotomus sergenti became infected with L. tropica. The average infection rate was 2.9%, and rats were infective for sand flies even 24 months post infection. The infectivity of the vertebrate host for insect vector was therefore not linked to the symptomatic stage of the infection. Such lack of correlation between clinical symptoms and infectivity to sand flies was reported previously for Leishmania infantum, the agent of visceral leishmaniasis; for species causing cutaneous leishmaniasis, however, this is the first evidence of transmission from a host without any visible cutaneous changes. If confirmed in the field, transmission from the asymptomatic host would be of great epidemiological significance. PMID- 12737991 TI - Characterisation of erythrocyte invasion by Babesia bovis merozoites efficiently released from their host cell after high-voltage pulsing. AB - Apicomplexa are a phylum of obligate intracellular parasites critically dependent on invasion of a host cell. An in vitro assay for erythrocyte invasion by Babesia bovis was established, employing free merozoites obtained after the application of high-voltage to the parasitised erythrocytes. The invasion proceeds efficiently in phosphate-buffered saline solution without the requirement for any serum or medium components. The kinetics of invasion can be measured over a time span of 5-60 min after which invasion is completed at an average efficiency of 41%. The fast kinetics and high efficiency exceed those of most previously established apicomplexan invasion assays. The manipulation of intracellular calcium concentration inhibits invasion. Preincubation of merozoites at 37 degrees C also reduces invasion, possibly by the premature secretion of protein. Proteins that are shed into the environment during invasion were directly detectable by protein staining after 2-D gel electrophoresis. The limitations posed by the immunological detection of proteins released during in vitro invasion by other apicomplexan parasites can, therefore, be avoided by this method. A unique feature of the assay is the reversible uncoupling of invasion and intracellular development, the latter taking place only under serum-rich medium conditions. In addition, host cell attachment is uncoupled from invasion by cytochalasin B. PMID- 12737992 TI - Virus multiplication and induction of apoptosis by Sendai virus: role of the C proteins. AB - Sendai virus (SeV) P gene encodes a nested set of carboxyl-coterminal proteins (C', C, Y1 and Y2), which are referred to collectively as the C proteins. Characterization of the virus multiplication and cellular responses in HEp-2 cells infected with the recombinant SeV which lacks two (C' and C), three (C', C and Y1) or all the four C proteins revealed that all the recombinant viruses can grow in the cells to various extents, depending, apparently, on the number of species expressing C protein. In reverse proportion to the viral growth ability, these viruses induced apoptosis in the infected cells. These results indicate that Y2 protein has an antiapoptotic activity, and suggest that this activity works in an additive manner with the longer C protein(s) (C' and/or C) of SeV in order to suppress virus-induced apoptosis in the SeV-infected cells. Apparently, the antiapoptotic activity of the C proteins supports virus multiplication in the infected cells. PMID- 12737993 TI - Serological reactivity of baculovirus-expressed Ebola virus VP35 and nucleoproteins. AB - Ebola virus (EBOV) is a member of the family Filoviridae and is classified as a biosafety level 4 virus. This classification makes the preparation of antigen and performance of diagnostic assays time-consuming and complicated. The objective of this study was to evaluate the value of EBOV immunoassays based on recombinant nucleoprotein (r-NP) and recombinant VP35 (r-VP35) using large serum panels of African origin and from primates. Furthermore, we investigated whether the results obtained with EBOV r-VP35 enzyme-linked immunosorbent assay (ELISA) could improve on the findings obtained with the EBOV r-NP ELISA. The full-length EBOV NP and VP35 of the EBOV subtype Zaire were expressed as histidine-tagged recombinant proteins in the baculovirus expression system. The antigenic reactivity and specificity of these recombinant proteins were determined by Western blotting and ELISA using EBOV specific monoclonal antibodies. The results obtained with the r-NP and r-VP35 ELISAs were compared with the results obtained in an indirect immunofluorescence assay based on native EBOV subtype Zaire. EBOV specific monoclonal antibodies reacted specifically with the respective proteins in both Western blot and ELISA. Five hundred and twenty six samples from humans and primates were tested with r-NP and r-VP35 ELISAs. Monkey serum samples positive for EBOV subtype Reston and Zaire were both positive in the EBOV r-NP ELISA, whereas only the EBOV Zaire infected monkeys were positive in the r-VP35 ELISA. The sensitivity and specificity values of the EBOV recombinants' ELISAs compared to those of the immunofluorescence assay were 92% and 99% for r-NP and 44% and 100% for r-VP35. r-NP ELISA proved to be a sensitive and specific assay for EBOV diagnosis and for epidemiological studies for both EBOV subtypes Reston and Zaire. The use of r-VP35 in an ELISA format has no additional value for EBOV serodiagnosis. PMID- 12737994 TI - Vpx and Vpr proteins of HIV-2 up-regulate the viral infectivity by a distinct mechanism in lymphocytic cells. AB - Mutants of human immunodeficiency virus type 2 (HIV-2) carrying a frame-shift mutation in vpx, vpr, and in both genes were monitored for their growth potentials in a newly established lymphocytic cell line, HSC-F. Worthy of note, the replication of a vpx single mutant, but not vpr, was severely impaired in these cells, and that of a vpx-vpr double mutant was more damaged. Defective replication sites of the vpx single and vpx-vpr double mutants were demonstrated to be mapped, respectively, to the nuclear import of viral genome, and to both, this process and the virus assembly/release stage. While the mutational effect of vpr was small, the replication efficiency in one cycle of the vpx mutant relative to that of wild-type virus was estimated to be 10%. The growth phenotypes of the vpx, vpr, and vpx-vpr mutant viruses in HSC-F cells were essentially repeated in primary human lymphocytes. In primary human macrophages, whereas the vpx and vpx vpr mutants did not grow at all, the vpr mutant grew equally as well as the wild type virus. These results strongly suggested that Vpx is critical for up regulation of HIV-2 replication in natural target cells by enhancing the genome nuclear import, and that Vpr promotes HIV-2 replication somewhat, at least in lymphocytic cells, at a very late replication phase. PMID- 12737996 TI - Vitamin A deficiency leads to severe functional disturbance of the intestinal epithelium enzymes associated with diarrhoea and increased bacterial translocation in gnotobiotic rats. AB - A disturbance of the integrity of the intestinal epithelium with an increased risk for bacterial translocation is one of the suggested factors underlying the increased incidence of infections and septicaemia during vitamin A deficiency. In the present study the effects of vitamin A deficiency on the enzymic activity of enterocytes in response to bacterial colonization with a non-pathogenic Escherichia coli strain were studied in monocolonized and conventional Wistar rats. The monocolonized, but not the conventional, vitamin A-deficient rats had markedly reduced weight compared to their pair-fed controls and presented neurological symptoms, such as hind leg weakness, tremor and slow gait. Moreover, only in the monocolonized vitamin A-deficient rats were severe diarrhoea and bacterial translocation to extraintestinal sites-mainly kidneys-detected. Measurements of enterocyte brush-border enzyme activities revealed that lactase, sucrase, gamma-glutamyltranspeptidase (GGT) and dipeptidyl peptidase IV (DPP IV) were significantly reduced in the monocolonized vitamin A-deficient rats compared to the pair-fed controls, indicating a severe functional disturbance of the enterocytes. In conventional vitamin A-deficient rats only sucrase activity was markedly lower than in the respective controls. Our observation, that the deficient vitamin A status led to a strong reduction of enterocyte enzymic activities, associated with diarrhoea and increased bacterial translocation, mainly in the gnotobiotic rats, suggests that the composition of the bacterial flora, i.e. the colonization state, has a strong influence on triggering the severity of the functional disturbances of the intestinal epithelium, and adds to the clinical manifestations of vitamin A deficiency. PMID- 12737995 TI - Francisella novicida LPS has greater immunobiological activity in mice than F. tularensis LPS, and contributes to F. novicida murine pathogenesis. AB - To further understand the role of LPS in the pathogenesis of Francisella infection, we characterized murine infection with F. novicida, and compared immunobiological activities of F. novicida LPS and the LPS from F. tularensis live vaccine strain (LVS). F. novicida had a lower intradermal LD(50) in BALB/cByJ mice than F. tularensis LVS, and mice given a lethal F. novicida dose intraperitoneally died faster than those given the same lethal F. tularensis LVS dose. However, the pattern of in vivo dissemination was similar, and in vitro growth of both bacteria in bone marrow-derived macrophages was comparable. F. novicida LPS stimulated very modest in vitro proliferation of mouse splenocytes at high doses, but F. tularensis LVS LPS did not. Murine bone marrow macrophages treated in vitro with F. novicida LPS produced IL12 and TNF-alpha, but did not produce detectable interferon-gamma, IL10, or nitric oxide; in contrast, murine macrophages treated with F. tularensis LVS LPS produced none of these mediators. In contrast to clear differences in stimulation of proliferation and especially cytokines, both types of purified LPS stimulated early protection against lethal challenge of mice with F. tularensis LVS, but not against lethal challenge with F. novicida. Thus, although LPS recognition may not be a major factor in engendering protection, the ability of F. novicida LPS to stimulate the production of proinflammatory cytokines including TNF-alpha likely contributes to the increased virulence for mice of F. novicida compared to F. tularensis LVS. PMID- 12737997 TI - Production of pro- and anti-inflammatory cytokines by monocytes from patients with paracoccidioidomycosis. AB - Monocytes and macrophages can produce a large repertoire of cytokines and participate in the pathogenesis of granulomatous diseases. We investigated the production of pro- and anti-inflammatory cytokines by monocytes from patients with active paracoccidioidomycosis. Peripheral blood monocytes from 37 patients and 29 healthy controls were cultivated with or without 10 microg/ml of lipopolysaccharide (LPS) for 18 h at 37 degrees C, and the cytokine levels were determined in the culture supernatants by enzyme immunoassay. The results showed that the endogenous levels of tumor necrosis factor alpha (TNF-alpha), interleukin-1beta (IL-1beta), IL-6, IL-8, IL-10 and transforming growth factor beta detected in the supernatant of patient monocytes cultivated without stimulus were significantly higher than those produced by healthy controls. These data demonstrated that monocytes from patients with active paracoccidioidomycosis produce high levels of cytokines with both inflammatory and anti-inflammatory activities. However, patient monocytes produced significantly lower TNF-alpha and IL-6 levels in response to LPS when compared to normal subjects, suggesting an impairment in their capacity to produce these cytokines after LPS stimulation. Concentrations of IL-1beta, IL-8 and IL-10 in cultures stimulated with LPS were higher in patients than in controls. These results suggest that an imbalance in the production of pro- and anti-inflammatory cytokines might be associated with the pathogenesis of paracoccidioidomycosis. PMID- 12737998 TI - Trypanosoma cruzi clonal diversity and the epidemiology of Chagas' disease. AB - Chagas' disease is caused by the protozoan Trypanosoma cruzi and it has a variable clinical outcome. The basis for this variability relies in part on the complexity of the parasite population consisting of multiple clones displaying distinct biological properties. A major current challenge is to correlate parasite genetic variability with pathogenesis. PMID- 12737999 TI - Corruption of host seven-transmembrane proteins by pathogenic microbes: a common theme in animals and plants? AB - Human diseases like AIDS, malaria, and pneumonia are caused by pathogens that corrupt host chemokine G-protein coupled receptors for molecular docking. Comparatively, little is known about plant host factors that are required for pathogenesis and that may serve as receptors for the entry of pathogenic microbes. Here, we review potential analogies between human chemokine receptors and the plant seven-transmembrane MLO protein, a candidate serving a dual role as docking molecule and defence modulator for the phytopathogenic powdery mildew fungus. PMID- 12738000 TI - Immune-mediated clearance of virus from the central nervous system. AB - Immune-mediated clearance of virus from the central nervous system (CNS) differs from that of the other organs. Mechanisms of virus control are largely dependent upon the target cell type. Although cytolytic T lymphocytes may mediate clearance of virus from glial cells, non-cytolytic mechanisms mediated by antibody and cytokines dominate clearance from neurons. PMID- 12738001 TI - Medical and economic impact of extraintestinal infections due to Escherichia coli: focus on an increasingly important endemic problem. AB - Escherichia coli is probably the best-known bacterial species and one of the most frequently isolated organisms from clinical specimens. Despite this, underappreciation and misunderstandings exist among medical professionals and the lay public alike regarding E. coli as an extraintestinal pathogen. Underappreciated features include (i) the wide variety of extraintestinal infections E. coli can cause, (ii) the high incidence and associated morbidity, mortality, and costs of these diverse clinical syndromes, (iii) the pathogenic potential of different groups of E. coli strains for causing intestinal versus extraintestinal disease, and (iv) increasing antimicrobial resistance. In this era in which health news often sensationalizes uncommon infection syndromes or pathogens, the strains of E. coli that cause extraintestinal infection are an increasingly important endemic problem and underappreciated "killers". Billions of health care dollars, millions of work days, and hundreds of thousands of lives are lost each year to extraintestinal infections due to E. coli. New treatments and prevention measures will be needed for improved outcomes and a diminished disease burden. PMID- 12738002 TI - Development of a vaccine for toxoplasmosis: current status. AB - The need for a vaccine for human toxoplasmosis is briefly discussed. Recent progress in knowledge of the protective immune response generated by Toxoplasma gondii and the current status of development of a vaccine for toxoplasmosis are highlighted. PMID- 12738004 TI - Optical recordings of Ca2+ signaling activities from identified inner ear cells in cochlear slices and hemicochleae. AB - One of the major obstacles hindering the progress of studies on mammalian cochlear physiology is the inaccessibility of inner ear cells located in a complex structure of the bony labyrinth. We describe here a technique to record cellular fluorescent signals from any identified inner ear cells in cochlear slices and hemicochleae. Cochlear slices were obtained from postnatal rats (P0 P7) before the cochlea completely ossify, and hemicochleae were cut from older animals (P7-adult). Individual inner ear cells were visually identified using infrared differential interference contrast or oblique illumination optics. Techniques were developed for either bulk-loading cells or loading selected single cells with Ca(2+) indicator dyes, and for maintaining functional viability of cochlear slices/hemicochleae for recordings. Robust and reliable responses of ligand-gated receptors were recorded from individual inner ear cells (e.g. hair cells, spiral ganglion neurons etc.) for at least 24 h after slices/hemicochleae were cut by an oscillating tissue slicer. The technique described here allowed direct observations of [Ca(2+)](i) activities from multiple cells simultaneously in situ, thus providing a feasible way to study the intercellular communication or networking activities from identified cells in the inner ear. PMID- 12738003 TI - Audition of laughing and crying leads to right amygdala activation in a low-noise fMRI setting. AB - Adequate behavioral responses to socially relevant stimuli are often impaired after lesions of the amygdala. These impaired behavioral responses in particular concern the recognition of facial, and sometimes vocal, expressions of fear. Using low-noise functional magnetic resonance imaging (fMRI) in combination with controlled sound delivery, we investigated how the amygdala, insula and auditory cortex are involved in the processing of affective non-verbal vocalizations (laughing, crying) in healthy humans. The same samples of male and female laughing and crying were presented in two different experimental conditions: self induction of the corresponding emotions while listening, and detection of artificial pitch shifts in the same stimuli. Both conditions led to bilateral activation of the amygdala, insula and auditory cortex with a right-hemisphere advantage in the amygdala, and larger activation during laughing than crying in the auditory cortex with a slight right-hemisphere advantage for laughing, both likely due to acoustic stimulus features. The results show that amygdala activation by emotionally meaningful sounds like laughing and crying is independent of the emotional involvement, suggesting the pattern recognition aspect of these sounds is crucial for this activation. This aspect was revealed by a low-noise fMRI protocol which presumably minimized confounding effects of stressful high-noise fMRI. PMID- 12738005 TI - Organotypic slices in vitro: repeated, same-cell, high-resolution tracking of nuclear and cytoplasmic fluorescent signals in live, transfected cerebellar neurons by confocal microscopy. AB - The culture of organotypic slices for the purposes of tracking dynamic cellular events within the same live cell at high resolution, as a function of development in vitro has not been previously reported. The present study was undertaken to define the conditions most suitable for both the in vitro organotypic development of Purkinje neurons in cerebellar slices of neonatal mice, and the repeated visualization of nuclear signals within such cells. Slices of cerebella were maintained on 25 mm diameter, collagen-coated Anodisc membranes, placed in six well plates and raised to the air-medium interface by use of glass fibre filter supports. This system permits cultures to be repeatedly observed both by phase contrast microscopy and, upon biolistic transfection, by laser confocal microscopy using 40x, 60x, and 100x water-immersion objectives, at high resolution. Upon co-transfection with two plasmids, differentiation of the same transfected Purkinje neurons was followed across in vitro development for periods of up to 10 days. Despite the relative thickness of the slice culture, even small, punctate, nuclear signals, were detectable. The results show that Purkinje neurons in cerebellar slices explanted from postnatal day 2 mice, developed cytotypically, although some were ectopically located. In contrast, Purkinje neurons in slices from postnatal day 6 cerebella developed in an organotypic manner. It is concluded that this culture system serves as an ideal tool for applications in experimental biology where high resolution tracking of cellular signals, over extended time periods, is of interest. PMID- 12738006 TI - In vitro phosphorylation of cytoskeletal proteins from cerebral cortex of rats. AB - Procedures for the preparation of high- and low-salt Triton insoluble cytoskeletal fractions from rat brain suitable for studying in vitro phosphorylation by endogenous kinases and phosphatases are described. The high salt Triton insoluble cytoskeletal fraction is enriched in neurofilament subunits (NF-H, NF-M and NF-L), vimentin and glial fibrillary acidic protein (GFAP), while the low-salt Triton insoluble cytoskeletal fraction contains detergent insoluble cytoskeletal elements such as intermediate filament subunits and tubulins. One of our approaches is to incubate cerebral cortex slices with [32P]orthophosphate before the cytoskeletal fraction extraction, which allows the in vitro phosphorylation of cytoskeletal constituents in an intact intracellular environment. On the other hand, we also incubate low- or high-salt cytoskeletal fractions previously prepared with [gamma(32)P]ATP. By doing so, we are able to study the direct effects of substances on the kinase and phosphatase activities associated with the cytoskeletal fraction. Moreover by using specific activators or inhibitors of protein kinases and phosphatases we can obtain more detailed information on the alterations provoked by these substances. These approaches are useful for the investigation of the neurotoxic effects of various drugs and metabolites affecting the cytoskeletal-associated phosphorylation system in the brain. PMID- 12738007 TI - A rapid assay for glial cell line-derived neurotrophic factor and neurturin based on transfection of cells with tyrosine hydroxylase promoter-luciferase construct. AB - Glial cell line-derived neurotrophic factor (GDNF), a potent survival and trophic factor for various neuronal cells, has been measured by assaying its bioactivity based on neurite outgrowth or cell proliferation. We describe herein a sensitive and simple non-radioactive quantitative bioassay for GDNF family proteins based on their ability to induce tyrosine hydroxylase (TH) gene expression. Human neuroblastoma SK-N-MC cells were stably transfected with expression constructs of c-ret and with a luciferase reporter gene driven by 2 kb of the rat TH gene promoter region. In the presence of GDNF, luciferase activity increased with 20 h of incubation. A dose-dependent increase in luciferase activity was observed in the presence of GDNF between 1 and 300 ng/ml. This assay was also applicable for the quantification of the bioactivity of neurturin, another member of the GDNF family showing an even more sensitive profile of dose dependency than GDNF. Typical culture media were applicable in this assay. This method can be easily applied to numerous samples of conditioned medium in a short time. PMID- 12738008 TI - Long-term stimulation of mouse hippocampal slice culture on microelectrode array. AB - To understand mechanisms of information processing, development and degeneration of the central nervous system, simultaneous multisite recording and stimulation have become extremely helpful. We have further developed the innovative approach to record from intact neural networks using planar microelectrode arrays (MEAs) with 60 substrate-integrated nano-columnar electrodes. To allow for long-term stimulation, mouse hippocampal tissue slices were immobilized onto MEAs and permanently moved between the gas and medium phase in a specifically designed tilting incubator that made it possible to electrically contact up to 90 MEAs with 5400 electrodes. After 2-3 weeks in vitro, histochemical staining, the intracellular microinjection of the fluorescent dye Alexa and the recording of spontaneous activity revealed in vivo-like characteristics of the organotypically cultured tissue. The feasibility of long-term stimulation during culturing was demonstrated with a low frequency paradigm. 0.003 Hz stimulation over a 16 h period resulted in a significant decline of field potentials and population spikes in two identified hippocampal subregions. Control experiments revealed that this effect was not due to tissue detachment or to induced cell death. In summary, the novel technology promises to open a new avenue for analyzing regulatory interactions of neuronal activity, cell differentiation and gene expression during development and in diseases. PMID- 12738011 TI - [Which future for medicine during catastrophe? Lessons from AZF factory catastrophe in Toulouse]. PMID- 12738009 TI - Two approaches to double post-embedding immunogold labeling of freeze-substituted tissue embedded in low temperature Lowicryl HM20 resin. AB - Double labeling is used for localizing two antigens simultaneously in the same tissue. We have used two approaches to post-embedding immunogold labeling to investigate whether nerve terminals in the guinea-pig anteroventral cochlear nucleus (AVCN) that contain gamma-aminobutyric acid (GABA) or glycine are capable of retrieving the other amino acid as part of an investigation of colocalization of these putative neurotransmitters. For this, vibroslices of perfusion-fixed brain stem were freeze-substituted and embedded in the low temperature resin, Lowicryl HM20. Simultaneous labeling of ultrathin sections was then performed with a mixture of a rabbit primary antibody to GABA and a guinea-pig primary antibody to the glycine transporter, GLYT2, followed by labeling with a mixture of secondary antibodies (goat anti-rabbit IgG-30 nm gold, goat anti-guinea pig IgG-15 nm gold). This approach indicated that GLYT2 occurs in the plasma membrane of some terminals that contain GABA. The other approach involved sequential labeling of ultrathin sections with a rabbit primary antibody to the GABA transporter, GAT1, followed by an anti-rabbit secondary antibody conjugated to 15 nm gold particles. Sections were then treated with paraformaldehyde vapor to denature any free anti-IgG binding sites on the first antibody, and labeled with a primary antibody to glycine also raised in rabbit followed by an anti-rabbit secondary antibody conjugated to 30-nm gold particles. This approach indicated that GAT1 occurs in the plasma membrane of some terminals that contain glycine. Thus, these techniques can be used to localize heat-labile multiple antigens in the same tissue. PMID- 12738012 TI - [Do we have to modify the "Plan Rouge" for management of casualties during disasters with limited amplitude?]. PMID- 12738013 TI - [Organization of medical rescue during catastrophes with limited effects occurring in urban areas]. AB - OBJECTIVE: We conducted a survey regarding the organization of medical rescue during major events (catastrophes with limited effects) occurring in urban areas (Paris and immediate suburbs). The goal of this work was to study the availability of on site medical help and the real needs.Study design - Retrospective survey. METHODS: Thirty-eight major events were analysed between 1988 and 2000. The median number (25th-75th percentiles) of victims per event was 42 (21-68) (range 8 to 424). RESULTS: The median percentage of true emergencies (TE) was 5% with regard to the total number of victims per event. Thirty minutes after the event, 92% of the sites had a number of physician-manned ambulances greater than the number of severe victims. The median time to first evacuation was 79 (62-102) min. CONCLUSION: Disasters with limited effect occurrence in Paris and its immediate suburbs are characterized by a small percentage of TE and by a constant oversupply of medical means onsite. These observations led us to propose a new organization of medical rescue during this type of catastrophe, abandoning the classical notion of forward medical command post (FMCP) for a collection point of medical services (CPMS) consisting all means of evacuation (physician-manned and other ambulances). Also, a new type of victim identification, based on hospital base-station medical direction is discussed in this paper. PMID- 12738014 TI - [The impact of a French guideline on the clinical management of severe head injury (SHI) published in 1999]. AB - OBJECTIVE: To evaluate the impact of a French guideline on the clinical management of severe head injury (SHI) published in 1999. STUDY DESIGN: Nationwide survey. METHODS: 182 medical centres answered the inquiry. RESULTS: 111 centres declared to manage such patients suffering from SHI. 68% of doctors from these clinical departments did report to have read these guidelines. Fifty percent of them found in these guidelines useful data for their clinical practice. Forty four percent of centres could not monitor ICP lacking neurosurgical facility in their hospital. Conversely, all hospitals with neurosurgery available did monitor ICP. Seventy six percent of centres reported difficulties to find a facility, which would take these patients in charge when discharged from ICU. We analysed the main items included in the guidelines and report their impact. Seventy four percent of responders found that clinical management of SHI patients had improved during the last years. CONCLUSION: The French guidelines had a significant impact and seem to have provided a useful aid to clinical management of SHI patients. ICP monitoring and the shortage in post ICU facilities are remaining issues. PMID- 12738015 TI - [Effects of equiosmolar load of 20% mannitol, 7.5% saline and 0.9% saline on plasma osmolarity, haemodynamics and plasma concentrations of electrolytes]. AB - OBJECTIVES: Mannitol 20% (M) and NaCl 7.5% (H) are hypertonic solutions used in patient in hypovolaemic shock and in case of increased ICP. To our knowledge no study in humans compared them in an equiosmolar load. The objectives of this study were to compare the pharmacokinetics of both solutions in an equiosmolar load with an isotonic solution (NaCl 0.9% (C)) in terms of osmolarity, electrolytes and haemodynamic parameters over time. STUDY DESIGN: Prospective, randomized, controlled study. PATIENTS: Thirty ASA I and II patients undergoing non haemorrhagic surgery under general anesthesia. METHODS: Three groups were studied (groups M, H and C), each receiving an equiosmolar load (4.95 mOsm kg( 1)) of solutions of mannitol 20%, saline 7.5% and saline 0.9% over ten minutes. Vital parameters (non invasive blood pressure, heart rate, oesophageal temperature) were monitored and blood samples (hemoglobin, hematocrit, K(+), Na(+), Cl(-), urea, creatinine, glucose, mannitolemia, osmolarity) were assessed preoperatively and intraoperatively at times 0, 3, 5, 10, 15, 30, 45, 60, 90, 120, 150 min of perfusion. RESULTS: In both hypertonic groups, osmolarity was maximal at the end of infusion (group M: 329 +/- 7 mOsm l(-1); group H: 321 +/- 11 mOsm l(-1)); at the same time, Na(+) value was lowest in group M: 129 +/- 3 mmol l(-1) and highest in group H: 151 +/- 5 mmol l(-1), P < 0.001) with normalisation at 60 min. These results were also statistically significant when compared to the isotonic group (group C: osmolarity: 296 +/- 3 mOsm l(-1), P < 0.001; Na(+): 140 +/- 2 mmol l(-1), P < 0.001). Plasma volume expansion was statistically significantly larger in-group C between 10 and 15 min compared to both hypertonic groups (haematocrit: group C: 35 +/- 4%, group M: 39 +/- 5%; group H: 41 +/- 3%, P < 0.004). Haemodynamic parameters were similar in the 3 groups with however a trend towards lowers HR in the mannitol group, particularly at T10. Temperature was lower in the control group because of the volume infused. Cl(-) was higher in the H group. CONCLUSION: A single infusion of 4.95 mOsm kg( 1) of mannitol 20% or NaCl 7.5% induces a similar osmolar variation over time with a maximal effect after 10 min. PMID- 12738017 TI - [Hepatorenal syndrome: from physiopathology to treatment]. AB - OBJECTIVES: Data synthesis on physiopathology and treatment of hepatorenal syndrome (HRS). DATA SOURCES: Data were searched in the Medline database from 1975 to 2002 using the following key-words: hepatorenal syndrome, ascite, cirrhosis and portal hypertension. DATA EXTRACTION: Publications from 1986 to 2002 were selected depending on the quality of their methodology and their pertinence. One publication from 1975 was kept. DATA SYNTHESIS: Hepatorenal syndrome is a common and severe complication of patients with advanced liver cirrhosis with ascites. It is a functional renal failure due to intense vasoconstriction of the renal circulation secondary to an intense splanchnic vasodilatation. Two types of HRS are differentiated mainly by the speed and the magnitude of the renal failure. Liver transplantation remains the best treatment but is rarely applicable because of the short survival after diagnosis. In the last few years, new therapy have been developed, vasoconstrictor drugs which mainly elicit their effects on the splanchnic circulation as vasopressin and principally its analogues ornipressine and terlipressine are effective in improving renal function and could act as bridge for liver transplantation. The place of the transjugular intrahepatic portosystemic shunt remain to be evaluated. CONCLUSION: Prognosis of patients with HRS remains poor but the pharmacologic treatment by terlipressine has improved the prognosis particularly in order to wait liver transplantation. PMID- 12738016 TI - [Intensive care management of 28 patients with severe eclampsia in a tropical African setting]. AB - OBJECTIVE: To study the specific management problems of severe eclampsia under tropical latitudes. STUDY DESIGN: A two years retrospective study in a University hospital in the tropics. PATIENTS AND METHODS: In all patients admitted for eclampsia between January 1997 et December 1999, the following parameters were studied: age, parity, interval between disease et admission, post-eclampsia Glasgow Coma Scale (GCS), time of occurrence of eclampsia during pregnancy, delivery route, blood pressure data at admission, the occurrence of complications at admission or during hospital stay. RESULTS: Twenty-eight mainly primiparous patients (mean age: 26 +/- 6) were admitted with an average delay of 8.5 +/- 10.2 hours after the first symptoms. The time of occurrence was prepartum in 6, perpartum in 14 and postpartum in 8 cases. All patients were hypertensive and comatose with an average GCS of 8 +/- 2.2. Twenty patients had been previously intubated and ventilated. Delivery was natural in 22 and by caesarean section in 6 patients. The following complications were found: acute oliguric renal failure (9), HELLP-syndrome (4), cerebral haemorrhage (4), acute lung oedema (3) and acute respiratory distress syndrome (1). Maternal and child mortality were 35 and 42.8% respectively. CONCLUSION: Eclampsia is a major cause of both maternal and infantile mortality in developing countries. The authors insist that prevention and management require speedy transfers to adapted specialized obstetrical intensive care structures. PMID- 12738018 TI - [Pneumomediastinum and bilateral tension pneumothorax as a complication of oxygen therapy using a nasal cannula]. AB - We report the case of a patient who developed a subcutaneous and submucosal emphysema in association with a pneumomediastinum, a bilateral pneumothorax and a pneumoperitoneum. This complication was secondary to oxygen supply via a nasal cannula, which allowed a wrong submucosal pathway previously, created by the traumatic placement of a nasogastric tube. The evolution was uneventful. We comment the pathophysiological mechanism of such a complication and propose simple actions to prevent the reproduction. PMID- 12738019 TI - [Internal carotid artery dissection after Heimlich maneuver]. AB - We report a case of cervical artery dissection following a Heimlich maneuver. Cervical artery dissections are at the present time well known and are sometimes associated with trivial traumas. However, to our knowledge, this complication of such maneuver was never reported in the literature. Pathophysiological mechanisms are discussed. PMID- 12738020 TI - [Convulsions due to a postpartum cerebral angiopathy associated with the administration of bromocriptine]. AB - We report a case of postpartum convulsions due to a benign cerebral angiopathy occurring after bromocriptine therapy to suppress lactation. This case analyses the current knowledge concerning this rare entity. It allows pointing out special measures that must be taken with the administration of bromocriptine. PMID- 12738021 TI - [Single ventricle and obstetric anaesthesia: two cases report]. AB - Single ventricle is a relatively rare abnormality found only in 0.5-1.5% of patients with congenital heart disease. Pregnancy results in an increased maternal morbidity and mortality in patients with right to left shunting, especially in cases associated with pulmonary hypertension and polycythemia and decreased arterial oxygen saturation. Most complications arise from decreases in systemic vascular resistance, causing an increase in right to left shunting and further hypoxemia and acidosis. For this reason, the major regional anaesthetic techniques are often avoided in these patients. The authors reported two successful management of regional anaesthesia for both caesarean and vaginal delivery. PMID- 12738022 TI - [Allergy investigations after two cases of adverse reactions to a neuromuscular blocking agent and management for subsequent general anaesthesia]. AB - We report two cases of severe anaphylactic reactions to rocuronium. Diagnosis was confirmed by skin tests and specific IgE assay. Cross-reactivity to all neuromuscular blocking agents was investigated by intradermal tests and leucocyte histamine release test. Intradermal tests and leukocyte histamine release were negative for cisatracurium. The two patients had undergone a subsequent general anaesthesia using cisatracurium and did not present any adverse reaction. PMID- 12738024 TI - [Is symptom of post-dural puncture syndrome reproductible in the same patient?]. PMID- 12738023 TI - [Iodine intoxication after subcutaneous irrigations of povidone iodine]. AB - Irrigation of povidone iodine considered as a safe and effective procedure, is frequently used for deep infections. We report a case of intoxication by iodine in a man of 68-year-old after subcutaneous irrigations of Betadine at a concentration of 20% for a subcutaneous infection of the thigh. Abnormalities of cardiac conduction, lactic acidosis, acute renal failure, hypocalcaemia and thyroid dysfunction were the manifestations of the intoxication confirmed by a very high level of total blood iodine and urine iodine. PMID- 12738025 TI - [Normality of ventilation-perfusion scan in pulmonary embolism due to a technical error]. PMID- 12738026 TI - [The anaesthesia of child seen by paediatrician]. PMID- 12738027 TI - [Antibioprophylaxis in abdominal surgery: opinion of the surgeon?]. PMID- 12738028 TI - [Ambulatory anaesthesia: questions about the preanaesthetic examination]. PMID- 12738029 TI - Heart disease, methamphetamine and AIDS. AB - Methamphetamine (MA) not only affects the nervous system but also has cardiac toxicity and immunosuppressive properties. This manuscript will provide support that there is a relationship between MA use and heart disease as well as immune dysfunction. The cardiovascular manifestations of acute MA use include tachycardia, atrioventricular arrhythmias, myocardial ischemia, myocardial ischemia and hypertension, resulting in cardiac lesions. Chronic use of MA causes cardiomyopathy including cellular infiltration, myocardial hypertrophy, myocardium rupture and fibrosis. The increased catecholamine levels are responsible for the cardiac lesions induced by MA. The additional problem with MA use is its potential to disrupt the immune system function leading to suppression of mitogen-stimulated lymphocyte, a reduction in circulating lymphocyte numbers and alternation T-lymphocyte cytokine secretion as well as B cell proinflammatory cytokine secretion. Concomitant MA use and Human Immunodeficiency Virus (HIV) infection not only enhances immunosuppression associated with HIV but also increases the heart disease occurrence with a coincidentally complication of AIDS or AIDS medications. PMID- 12738030 TI - Induction of CYP3A4 by 1alpha,25-dyhydroxyvitamin D3 in HepG2 cells. AB - CYP3A4, the predominant cytochrome P450 (CYP) expressed in human liver, contributes to the metabolism of approximately half the drugs in use today. In general, human-derived cell lines fail to express CYPs. It was previously shown that CYP3A4 mRNA and CYP3A immunoreactive protein are induced by 1alpha,25 dyhydroxyvitamin D(3) (1alpha,25-(OH)(2)D(3)) in the human colon carcinoma cell line Caco-2. The aim of the present study was to examine whether 1alpha,25 (OH)(2)D(3) regulates CYP3A4 gene expression in HepG2 cells, a human hepatocarcinoma cell line. Treatment with 1alpha,25-(OH)(2)D(3) resulted in an induction of CYP3A4 mRNA and CYP3A4 immunoreactive protein, 1.5-fold and 4.0-fold respectively, when compared to control cultures, in a time-dependent fashion. These observations are in agreement with previous reports suggesting a role of 1alpha,25-(OH)(2)D(3) on CYP3A4 transcription regulation, and demonstrate that this hormone, as in Caco-2 cells, increase CYP3A4 levels in HepG2 cells. In conclusion, HepG2 cell cultures treated with 1alpha,25-(OH)(2)D(3), provides a useful model to study the function of CYP3A4 and its role in drug liver metabolism. PMID- 12738031 TI - Dipeptidyl peptidase IV (DPPIV) enzyme activity on immature T-cell line R1.1 is down-regulated by dynorphin-A(1-17) as a non-substrate inhibitor. AB - In this study we examined surface expression of CD26 and the corresponding enzyme activity of dipeptidyl peptidase IV (DPPIV) on the cells of immature murine T cell line, R1.1. The data obtained have shown that R1.1 cells express high density of surface CD26 as compared to normal thymus cells. This was associated with strong enzyme activity, which, based on substrates and inhibitor specificity, corresponded to DPPIV. The DPPIV enzyme activity of R1.1 cells was 10 times stronger than that found on normal murine thymus cells (V(max) = 39 micromol/min/10(6) cells, vs 3.7 micromol/min/10(6) cells, respectively). Upon activation with anti-CD3, up-regulation of both membrane CD26, as well as of DPPIV enzyme activity on R1.1 cells were observed. The finding of strong DPPIV on R1.1 cells makes them suitable model for testing putative substrates/inhibitors of the enzyme in its natural microenvironment. Since in addition to strong DPPIV, R1.1 cells also express kappa opioid receptors (KOR) [European Journal of Pharmacology 227 (1992) 257], we tested the effect of dynorphin-A(1-17), an endogenous opioid peptide with KOR selectivity, on DPPIV of R1.1 cells. Dynorphin A(1-17) down-regulated DPPIV in a dose-dependent manner, with the potency similar to that of substance P, a known natural DPPIV substrate [Journal of Pharmacology and Experimental Therapeutics 260 (1992) 1257]. DPPIV down-regulation was resistant to bestatin and thiorphan, the inhibitors of two cell surface peptidases (APN and NEP, respectively) with potential of dynorphin-A(1-17) degradation, suggesting that the mechanism underlying the observed effect does not involve degradative products of dynorphin-A(1-17). DPPIV down-regulation was also resistent to KOR antagonist, NBI, suggesting that the mechanism underlying the observed phenomenon involves neither cointernalization of KOR and DPPIV. Collectively, cells of immature T cell line, R1.1 exert strong DPPIV enzyme activity, which could be down-regulated in the presence of dynorphin-A(1-17) by mechanism that presumably includes non-substrate inhibition. By down-regulating DPPIV, dynorphin-A(1-17) may indirectly affect activity and/or specificity of natural substrates of DPPIV, such as substance P, RANTES, and endomorphins. PMID- 12738032 TI - Screening of medicinal plant extracts for antioxidant activity. AB - The methanol extracts of nine medicinal plants traditionally used in Chinese medicine were screened for antioxidant activity versus resveratrol, which has been shown to protect cells from oxidative damage [Toxicol. Lett. 102 (1998) 5]. Most of the plant extracts used in this study inhibited the H(2)O(2)-induced apoptosis of Chinese hamster lung fibroblast (V79-4) cells. The extracts of Areca catechu var. dulcissima, Paeonia suffruticosa, Alpinia officinarum, Glycyrrhiza uralensis and Cinnamomun cassia strongly enhanced viability against H(2)O(2) induced oxidative damage in V79-4 cells. Relatively high levels of 1,1-diphenyl-2 picrylhydrazyl (DPPH) radical scavenging activity were detected in extracts of Areca catechu var. dulcissima, Paeonia suffruticosa and Cinnamomun cassia (IC(50) < 6.0 microg/ml). The activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) were dose-dependently enhanced in V79-4 cells treated with most of the plant extracts. The extracts of Areca catechu var. dulcissima showed higher antioxidant activity than resveratrol in all experiments. These results suggest that the plant extracts prevent oxidative damage in normal cells probably because of their antioxidant characteristics. PMID- 12738034 TI - Identification of the binding sites and selectivity of sarpogrelate, a novel 5 HT2 antagonist, to human 5-HT2A, 5-HT2B and 5-HT2C receptor subtypes by molecular modeling. AB - The aim of the present study was to investigate the binding sites interactions and the selectivity of sarpogrelate to human 5-HT(2) receptor family (5-HT(2A), 5 HT(2B) and 5-HT(2C) receptor subtypes) using molecular modeling. Rhodopsin (RH) crystal structures were used as template to build structural models of the human serotonin-2A and -2C receptors (5-HT(2A)R, 5-HT(2C)R), whereas for 5-HT(2B)R, we used our previously published three-dimensional (3D) models based on bacteriorhodopsin (BR). Sarpogrelate, a novel 5-HT(2)R antagonist, was docked to the receptors. Molecular dynamics (MD) simulations produced the strongest interaction for 5-HT(2A)R/sarpogrelate complex. Upon binding, sarpogrelate constraints aromatic residues network (Trp(3.28), Phe(5.47), Trp(6.48), Phe(6.51), Phe(6.52) in 5-HT(2A)R; Phe(3.35), Phe(6.51), Trp(7.40) in 5-HT(2B)R; Trp(3.28), Phe(3.35), Phe(5.47), Trp(6.48), Phe(6.51), Phe(6.52) in 5-HT(2C)R) in a stacked configuration, preventing activation of the receptor. The models suggest that the structural origin of the selectivity of sarpogrelate to 5 HT(2A)R vs both 5-HT(2B)R and 5-HT(2C)R comes from the following results: (1) The tight interaction between the antagonist and the transmembrane domain (TMD) 3. Asp(3.32) neutralizes the cationic head and interacts simultaneously with carboxylic group hydrogen of the antagonist molecule. (2) Due to steric hindrance, Ser(5.46) (vs Ala(5.46) in 5HT(2B) and 5HT(2C)) prevents sarpogrelate to enter deeply inside the hydrophobic core of the helix bundle and to interact with Pro(5.50). (3) The side chain of Ile(4.56) (vs Ile(4.56) in 5HT(2B)R and Val(4.56) in 5HT(2C)R) constraints sarpogrelate to adjust its position by translating toward the strongly attractive Asp(3.32). These results are in good agreement with binding affinities (pKi) of sarpogrelate for 5-HT(2) receptor family expressed in transfected cell. PMID- 12738033 TI - Amomum xanthoides extract prevents cytokine-induced cell death of RINm5F cells through the inhibition of nitric oxide formation. AB - We previously showed that Amomum xanthoides extract prevented alloxan-induced diabetes through the suppression of NF-kappaB activation. In this study, the preventive effects of A. xanthoides extract on cytokine-induced beta-cell destruction were examined. Cytokines produced by immune cells infiltrating pancreatic islets are important mediators of beta-cell destruction in insulin dependent diabetes mellitus. A. xanthoides extract completely protected interleukin-1beta (IL-1beta) and interferon-gamma (IFN-gamma)-mediated cytotoxicity in rat insulinoma cell line (RINm5F). Incubation with A. xanthoides extract resulted in a significant reduction in IL-1beta and IFN-gamma-induced nitric oxide (NO) production, a finding that correlated well with reduced levels of the inducible form of NO synthase (iNOS) mRNA and protein. The molecular mechanism by which A. xanthoides extract inhibited iNOS gene expression appeared to involve the inhibition of NF-kappaB activation. Our results revealed the possible therapeutic value of A. xanthoides extract for the prevention of diabetes mellitus progression. PMID- 12738035 TI - Alpha-2-adrenoceptor hyporesponsiveness in isolated tissues of cholestatic animals: involvement of opioid and nitric oxide systems. AB - In the present study, the status of alpha(2)-adrenoceptors during cholestasis was investigated by the inhibitory effect of clonidine on the electrically stimulated contractions of mice vas deferens (MVD) and guinea pig ileum (GPI). Clonidine inhibited the contractions in both tissues in a dose-dependent manner. Compared to unoperated animals, there was a significant right-shift in the clonidine concentration-curves of both tissues obtained from 5-day bile-duct ligated (BDL) animals (p < 0.01), implying the hyporesponsiveness of alpha(2)-adrenoceptors during cholestasis. Chronic treatment with naltrexone (3 mg/kg/day) reversed the right-shift induced by cholestasis in both tissues. Administration of N(omega) nitro-L-arginine methyl ester (20 mg/kg/day) also partially reversed cholestasis induced effect on IC(50) of clonidine. These two treatments had no effect on IC(50) of tissues from controls. Chronic yohimbine treatment (5 mg/kg/day) recovered the effect of cholestasis on MVD, but sensitized the ileum of unoperated and BDL guinea pigs to clonidine to a similar extent, providing evidence for the role of the augmented adrenergic state of cholestasis in the hyporesponsiveness of norepinephrine-releasing neurons of MVD. We concluded that cholestasis is associated with the decreased responsiveness of alpha(2) adrenoceptors and the cholestasis-associated augmented opioidergic tone and increased NO production contribute to this process. PMID- 12738036 TI - Effects of 6-formylpterin as an internal source of hydrogen peroxide on cell death of human peripheral blood leukocytes. AB - The intracellular generation of hydrogen peroxide (H(2)O(2)) by 6-formylpterin and its effects on the cell surface exposure of phosphatidylserine (PS) as a marker of cell death were examined in human peripheral blood leukocytes, and the effects were compared with those of exogenously administered H(2)O(2). Neutrophils, monocytes and lymphocytes were isolated from fresh blood, and cultured for 24 h in vitro. In neutrophils, the intracellular H(2)O(2) generation was observed when the cells were incubated with 100-500 microM 6-formylpterin, and the PS exposure due to spontaneous apoptosis was inhibited. The underlying mechanism of the inhibition was attributed to the suppression of both activation and activity of caspase-3. On the other hand, exogenously administered 100 microM H(2)O(2) did not affect the PS exposure. The intracellular H(2)O(2) generation was also observed in monocytes and lymphocytes. In monocytes, 500 microM 6 formylpterin induced more PS exposure than 100 microM H(2)O(2) did. In lymphocytes, up to 500 microM 6-formylpterin did not induce conspicuous PS exposure, while 100 microM H(2)O(2) induced severe PS exposure. These findings indicated that the resistance against an internal and external source of H(2)O(2) are different among leukocytes, for example, lymphocytes are poorly resistant against external H(2)O(2) but highly resistant against internal one. PMID- 12738037 TI - Opioid activity of C8813, a novel and potent opioid analgesic. AB - Compound trans-4-(p-bromophenyl)-4-(dimethylamino)-1-(2-thiophen-2-yl-ethyl) cyclohexanol (C8813), structurally unrelated to morphine, is a novel analgesic. The present study examined the antinociception, opioid receptor selectivity and in vitro activity of C8813. The antinociceptive activity was evaluated using mouse hot plate and acetic acid writhing tests. In mouse hot plate test, the antinociceptive ED(50) of C8813 was 11.5 microg/kg, being 591 times and 3.4 times more potent than morphine and fentanyl respectively. In mouse writhing test, the antinociceptive ED(50) of C8813 was 16.9 microg/kg, being 55 times and 2.3 times more active than morphine and fentanyl respectively. In the opioid receptor binding assay, C8813 showed high affinity for mu-opioid receptor (K(i) = 1.37 nM) and delta-opioid receptor (K(i) = 3.24 nM) but almost no affinity for kappa opioid receptor (at 1 microM). In the bioassay, the inhibitory effect of C8813 in the guinea-pig ileum (GPI) was 16.5 times more potent than in the mouse vas deferens (MVD). The inhibitory effects of C8813 in the GPI and MVD could be antagonized by mu-opioid receptor antagonist naloxone and delta-opioid receptor antagonist ICI174,864 respectively. However, the inhibitory effect of C8813 in the rabbit vas deferens was very weak. These results indicated that C8813 was a potent analgesic and a high affinity agonist for the mu- and delta-opioid receptors. PMID- 12738038 TI - Tetrandrine-induced cell cycle arrest and apoptosis in Hep G2 cells. AB - The effects of tetrandrine in the human hepatoblastoma G2 (Hep G2) cell line were investigated in this study. The results showed that tetrandrine not only inhibited Hep G2 growth but also induced apoptosis and blocked cell cycle progression in the G1 phase. ELISA assay demonstrated that tetrandrine significantly increased the expression of p53 and p21/WAF1 protein, which caused cell cycle arrest. An enhancement in Fas/APO-1 and its two form ligands, membrane bound Fas ligand (mFasL) and soluble Fas ligand (sFasL), might be responsible for the apoptotic effect induced by tetrandrine. Taken together, p53 and Fas/FasL apoptotic system possibly participated in the antiproliferative activity of tetrandrine in Hep G2 cells. PMID- 12738039 TI - A systematic review of manic and depressive prodromes. AB - BACKGROUND: This paper explores whether individuals with a mood disorder can identify the nature and duration of depressive and manic prodromes. METHODS: Seventy-three publications of prodromal symptoms in bipolar and unipolar disorders were identified by computer searches of seven databases (including MEDLINE and PsycLIT) supplemented by hand searches of journals. Seventeen studies (total sample=1191 subjects) met criteria for inclusion in a systematic review. RESULTS: At least 80% of individuals with a mood disorder can identify one or more prodromal symptoms. There are limited data about unipolar disorders. In bipolar disorders, early symptoms of mania are identified more frequently than early symptoms of depression. The most robust early symptom of mania is sleep disturbance (median prevalence 77%). Early symptoms of depression are inconsistent. The mean length of manic prodromes (>20 days) was consistently reported to be longer than depressive prodromes (<19 days). However, depressive prodromes showed greater inter-individual variation (ranging from 2 to 365 days) in duration than manic prodromes (1-120 days). LIMITATIONS: Few prospective studies of bipolar, and particularly unipolar disorders have been reported. CONCLUSIONS: Early symptoms of relapse in affective disorders can be identified. Explanations of the apparent differences in the recognition and length of prodromes between mania and bipolar depression are explored. Further research on duration, sequence of symptom appearance and characteristics of prodromes is warranted to clarify the clinical usefulness of early symptom monitoring. PMID- 12738040 TI - Anxiety and impulsivity levels identify relevant subtypes in adolescents with at risk behavior. AB - BACKGROUND: Impulsivity (I) and anxiety (A) were hypothesized to be crucial clinical features in adolescents with at-risk behavior. We therefore classified them into sub-groups according to these two major dimensions. The study examined the relevance of these groups by describing their major diagnoses and behavioral characteristics. METHODS: During a 1-year period, all in-patients consecutively admitted for at-risk behavior, except those with a previous psychotropic treatment and/or schizophrenic disorders, were rated for anxiety and impulsivity, and categorized into four groups: impulsive and anxious (IA), impulsive and non anxious (Ia), non-impulsive and anxious (iA), non-impulsive and non-anxious (ia). We assessed the main behavioral disturbances (suicide attempt, carving, violence, delinquency, substance abuse, and eating disorder) and the main current axis I disorder in each sub-group. RESULTS: A total of 69 patients were included. In the IA group 62% exhibited hypomanic episodes and 87% recurrent suicide attempts. In the Ia group all exhibited conduct disorders, 93% were males, 80% delinquents, and 100% violent with others. Both groups reported a high percentage of cannabis use (67%). The iA group exhibited anorexia nervosa (73%) with a major depressive episode. The ia patients were mainly non-violent, first suicide attempts with low risk. LIMITATIONS: Long-term data are needed to assess the stability of these groups. CONCLUSIONS: We found that sub-typing adolescents with at-risk behavior into four groups according to their level of anxiety and impulsivity was highly predictive of being suicidal with mood disorders (AI), delinquent with conduct disorder (Ia), anorectic or depressed (Ai), and with substance abuse associated only to impulsivity. It is likely that this sub-typing of patients may be useful for prevention and therapeutics. The impulsive-anxious group (IA) appears closely related to the soft bipolar spectrum. A replication and follow-up data are now needed. PMID- 12738042 TI - Psychosocial and vascular risk factors of depression in later life. AB - BACKGROUND: Research on the aetiology of late-life depression has typically focused on either risk factors from the psychosocial stress-vulnerability domain or degenerative biological changes (for instance, vascular disease). We examined whether vascular risk factors could be interpreted within the stress vulnerability model of depression. METHODS: The data came from a case-control design, nested in a community survey of elderly people. We compared 83 persons with a recently started episode of major or minor depression with 83 controls, with respect to the occurrence of stressful life events, long-term difficulties, neuroticism, and vascular risk factors (hypertension, heart disease, stroke). RESULTS: The (non-significant) association of vascular risk factors and onset of depressive episodes was not modified by neuroticism or the presence of long-term difficulties. Quite unexpectedly, vascular risk factors seemed to neutralize the depressogenic effect of stressful life events. The effect of vascular risk was significantly stronger in depressive episodes not preceded by a life event than in onsets following an event. CONCLUSIONS: Vascular risk factors cannot be interpreted within the stress-vulnerability model, but represent another pathway to depression. Why vascular risk factors reduce the depressogenic effect of stressful life events is not clear. Replication is critical to exclude the possibility of a chance finding. PMID- 12738041 TI - Suicidal depressed vs. non-suicidal depressed adolescents: differences in recent psychopathology. AB - BACKGROUND: Few studies have focused on the differences between two depressed groups of patients in child psychiatry: the suicidal and the non-suicidal adolescent population. As in other countries, depression is one of the most prevalent diagnoses in adolescents in Hungary. AIMS: The present study was designed to determine (a) whether there are specific symptoms to differentiate between two clinical samples of depressed children: patients expressing suicidal behaviour and their peers with no suicide attempts, and (b) if there are significant differences between parents' and adolescents' reports of the same symptoms. METHODS: Using a recently developed semi-structured interview (Diagnostic Evaluation Schedule for Children and Adolescents-Hungarian version, ), 132 symptoms were assessed for two clinical groups of depressed adolescents: a suicidal group (N=54) [corrected], and a non-suicidal group (N=78) [corrected]. The suicidal group had all made an unsuccessful suicide attempt and/or had had frequent suicidal thoughts during the 6 months prior to the study. The non suicidal group had neither attempted suicide, nor had had suicidal thoughts during the previous 6 months. All cases were selected from a larger sample of 490 consecutively referred new outpatient children over a 1 year period in five psychiatric facilities in Hungary. Only 13-17-year-old adolescents participated in the study. Both samples were identified using operationalised computer algorithm criteria of DSM-IV major depressive disorder episode (MDD) irrespective of the current clinical diagnosis of the patients. The Pearson Chi-square test with Monte Carlo correction was used to evaluate the differences between the suicidal and the non-suicidal depressed samples. RESULTS: Hopelessness, negative self-esteem and violent behaviour were the only significant discriminators between the two study groups according to the parent interviews, with increased problem scores in the suicidal sample compared to the non-suicidal sample. Suicidal depressed adolescents view themselves as more depressed and violent than do non-suicidal depressed individuals and were less anxious about their parents. CONCLUSIONS: The two depressed samples (suicidal vs. non-suicidal individuals) have only very few dissimilarities. There are, however, some essential differences between the parental and adolescent perceptions of the suicidal and depressive symptoms of the adolescent. The findings of the study underscore the necessity of collecting data from both the parent and the adolescent. LIMITATIONS: Cross-sectional, no lifetime psychopathology, referred samples, no blind estimation of the suicidal status of patients. PMID- 12738043 TI - Cognitive, interpersonal, and behavioral predictors of patients' and spouses' depression. AB - BACKGROUND: This study examined the relationship between five theory-based variables and the severity of depression. The variables were self-concept, self and spouse ascribed hostility, spouses' level of depression, and involvement in home activities. METHODS: Based on DSM-IV criteria and Beck Depression Inventory (BDI) scores, 75 wife-depressed couples and 59 husband-depressed couples were included in the study. In addition to the BDI, both spouses completed questionnaires relating to self-concept, self- and spouse ascribed hostility, and involvement in home activities. RESULTS: Multiple regression analyses for male and female patients and spouses revealed that in the four groups self-concept was significantly related to the severity of depression. Irrespective of clinical status, involvement in home activities was significantly associated with the severity of depression of women. The relationship between spouse ascribed hostility and the level of depression of both spouses in husband-depressed couples approached significance so did the relationship between patient's and spouse's level of depression in wife-depressed couples. In husband-depressed couples the association between spouses' level of depression reached significance. CONCLUSIONS: The shared and gender-related predictors of severity of depression support an integrative theoretical approach to depression and offer practical implications. PMID- 12738044 TI - Suicide and general elections in Austria: do preceding regional suicide rate differentials foreshadow subsequent voting behavior swings? AB - BACKGROUND: Suicide-epidemiological research on short-term effects of elections on national/regional suicide and parasuicide incidence has yielded contradictory evidence. Reversing the cause-effect relationship of this line of research we investigated whether preceding regional suicide rates are related to subsequent election results. METHODS: For Austria's 121 districts, we regressed averaged standardized suicide rates for the preceding period (1988-1994) on political parties' subsequent electoral gains/losses (1999-to-1995) while controlling for a set of 12 domain-relevant psychosocial/economic indices. RESULTS: Stepwise weighted multiple regression led to a significant model. The 1999-to-1995 electoral gains/losses of two opposition parties, together with the population variation caused by migration balance and by births/deaths balance, accounted for a substantial part (30%) of the variability in preceding district-level suicide rates. Various other social indices failed to contribute further substantial increments to this model. CONCLUSIONS: This finding suggests that variations in preceding regional suicide incidence might be mirrored in subsequent changes in voting behavior. A speculative post hoc explanation for the finding is offered: on a community level, suicide's aftermath might produce socially and politically alienated survivors of suicide who co-shape swings towards opposition parties in subsequent general elections. The finding calls for more research on suicide's long-term aftermath. LIMITATIONS: Within-country replicability and cross-national generalizability await further investigation. At present, the factor/mechanism accounting for this finding is neither well-established nor has been directly tested. PMID- 12738045 TI - Population attributable risk of major depression for suicidal ideation in a random and representative community sample. AB - BACKGROUND: The importance of depression in suicidal behaviour and suicidal ideation is usually determined on clinical samples. However, public health planning also requires population data. This study utilised the population attributable risk statistic in determining the importance of major depression as a contributing factor to suicidal ideation in a random and representative sample of the population. METHOD: Major depression and suicidal ideation as well as demographic and clinical data were delineated in a random and representative population sample of 3010 subjects. The population attributable risk statistic was used to determine the contribution of major depression to suicidal ideation. RESULTS: Multivariate analysis demonstrated that major depression was the major contributor to the risk for suicidal ideation with a population attributable risk of 56.6%. CONCLUSIONS: These results, utilising different measures of depression and suicidal ideation to those few previous population attributable risk studies examining this issue, confirm the overwhelming importance of major depression as a contributing factor to suicidal ideation in the community. PMID- 12738046 TI - Criminality in the offspring of antenatally depressed mothers: a 33-year follow up of the Northern Finland 1966 Birth Cohort. AB - BACKGROUND: No epidemiological studies have been reported on the association between mothers' antenatal depression and criminality in their offspring. METHODS: The material consists of a general population cohort of 12059 children born in 1966 in Northern Finland and followed to the end of 1998. Mothers were asked at midgestation by a nurse at the antenatal clinic if they felt themselves to be depressed. The Finnish Ministry of Justice provided information for all descendants on criminal offences. The associations between maternal depression and subgroups of violent and non-violent, violent recidive and non-recidive criminality in male and female offspring were analysed. RESULTS: Of the mothers 14% had depressed mood during pregnancy. A total of 607 (10.9%) male and 72 (1.3%) female criminal offenders were included in the cohort. When adjusted for mother's marital status, smoking, wantedness of the pregnancy, place of residence, socioeconomic status and perinatal complications, the odds ratio (OR) for males was 1.4 (95% CI 1.0-1.9) in the association between maternal depression and non-violent offenders, 1.6 (1.1-2.4) between maternal depression and violent offenders and 1.7 (1.0-3.0) between maternal depression and violent recidivists. The adjusted OR for females involved in non-violent crimes was 1.7 (0.9-3.3) and 0.6 (0.1-6.0) for violent crimes. LIMITATIONS: Maternal depression was not checked using a standardized assessment. CONCLUSIONS: For male offspring of antenatally depressed mothers there was a significant but slight increase in criminality. The emotional state of a pregnant mother may have some, but limited, influence on later criminality in the offspring. PMID- 12738047 TI - Relevance of the catatonic syndrome to the mixed manic episode. AB - BACKGROUND: Catatonic symptoms have been associated with mixed mania in the older psychiatric literature, however, to date no systematic studies have been performed to assess their frequency in these patients. METHOD: Ninety-nine patients with bipolar disorder manic or mixed episode were assessed for the presence of catatonia. RESULTS: Thirty-nine patients fulfilled criteria for mixed mania of whom 24 were catatonic. Among the patients with pure mania, only three were catatonic. Eighteen catatonic patients with mixed mania required admission to the acute care unit (ACU). LIMITATIONS: Our findings only apply to severely ill patients with mixed mania who require ACU admission. Nevertheless, it is important to know, that the likelihood of overlooking catatonia in less severely ill patients with mixed mania is low and that it does not need to be routinely assessed on a general ward. CONCLUSIONS: Catatonia is frequent in mania and linked to the mixed episode. Catatonia in mixed mania is likely to be found among the severely ill group of patients with mixed mania, who require emergency treatment. PMID- 12738048 TI - Do clinicians understand why individuals stop taking lithium? AB - BACKGROUND: Previous research highlights that, over a 2-year period, up to 50% of individuals receiving lithium prophylaxis are non-adherent with medication against medical advice. The main reasons identified by clinicians only partially reflect the reasons given by patients. METHODS: Seventy-two subjects who were identified from biochemistry laboratory data as receiving lithium for an affective disorder and who gave written informed consent, completed the 'Reasons for Stopping Medication' questionnaire. Forty-one clinicians involved in treating these 72 patients also completed the same questionnaire. Ratings on the 'Reasons for Stopping Medication' questionnaire were compared between adherent and non adherent patients and between patients and their clinicians. RESULTS: Adherent (n=39) and previously non-adherent patients (n=33) showed a high level of concordance in their rank ordering of reasons for considering stopping taking lithium (Kendall's W=0.059; P=0.001), with 'bothered by the idea that moods were controlled by medication', 'bothered by the idea of a chronic illness' and 'felt depressed' being the most commonly endorsed items. Clinicians and patients views showed low levels of concordance (Kendall's W=0.019; P=0.32). Clinicians identified 'missing highs', and 'felt well, saw no need to take medication' as the most likely reasons for non-adherence. The differences in the primary reasons for stopping lithium identified by clinicians and patients were statistically significant (chi(2)=61.1; df 40, P=0.005). CONCLUSIONS: Patients' reasons for stopping lithium appear to be influenced by concerns about what having an affective disorder and taking medication says about them. Clinicians are not fully aware of the main reasons patients would stop prophylactic treatment. PMID- 12738049 TI - Auditory event-related potentials in 50 melancholic patients: increased N100, N200 and P300 latencies and diminished P300 amplitude. AB - BACKGROUND: To investigate whether there are some differences in Event-Related Potentials (ERP) between melancholic patients and healthy controls. To establish whether there is a relationship between abnormalities of ERP and severity of depression and psychomotor retardation. METHOD: Melancholic depressed patients (N=50) and normal comparison subjects (N=31) were assessed for latencies and interlatencies of N100, N200, N400, latency and amplitude of P300. The ERPs were studied with an 'oddball paradigm' in the auditory modality. Severity of depression was measured by the Hamilton Depression Rating Scale (HDRS) and psychomotor retardation with the Depressive Retardation Rating Scale (DRRS). RESULTS: The melancholic group showed a significantly higher latency in N100 (P<0.001), N200 (P<0.001) and P300 (P<0.001) and a significantly lower P300 amplitude (P<0.001) than healthy controls. No other differences were found either in the latencies of the N400 or in their interlatencies. HDRS and DRRS do not have any significant correlations with amplitude or latency measures. LIMITATIONS: The subjects of this study are inpatients, with a severe subcategory of depression and high average age. It is difficult to generalize these findings. CONCLUSIONS: The principal finding of this study is the increase in three of the four latencies measured (N100, N200 and P300) and in the decreased P300 amplitude in melancholic patients compared to normal controls. There is no association between these abnormalities and clinical variables. PMID- 12738050 TI - Depression in women with polycystic ovary syndrome: clinical and biochemical correlates. AB - BACKGROUND: We assessed the prevalence of mood disturbance among women with prospectively documented polycystic ovary syndrome (PCOS). METHODS: Thirty-two women with PCOS completed the Center for Epidemiological Studies-Depression Rating Scale (CES-D). Clinical and biochemical characteristics were assessed. RESULTS: Sixteen women had CES-D scores indicative of depression. Depression was associated with greater insulin resistance (P=0.02) and higher body mass index (P=0.05). Women receiving oral contraceptives for the treatment of PCOS were less depressed than patients not receiving treatment (P=0.03). LIMITATIONS: Possible selection bias, use of a screening tool alone without further diagnostic evaluation of depression, small samples size and lack of direct comparison with an age matched control group, should be considered in interpretation of these results. CONCLUSION: Findings suggest a high prevalence of depression among women with PCOS, and an association between depression and PCOS markers. PMID- 12738052 TI - Symptoms of seasonal affective disorder and of obsessive-compulsive disorder reduced by light therapy. PMID- 12738051 TI - The effects of tryptophan depletion on mood and psychiatric symptoms. PMID- 12738053 TI - T-cell immune responses in the brain and their relevance for cerebral malignancies. AB - In order that cellular immune responses afford protection without risk to sensitive normal tissue, they must be adapted to individual tissues of the body. Nowhere is this more critical than for the brain, where various passive and active mechanisms maintain a state of immune privilege that can limit high magnitude immune responses. Nevertheless, it is now clear that immune responses are induced to antigens in the brain, including those expressed by cerebral malignancies. We discuss hypotheses of how this can occur, although details such as which antigen presenting cells are involved remain to be clarified. Antitumor responses induced spontaneously are insufficient to eradicate malignant astrocytomas; many studies suggest that this can be explained by a combination of low level immune response induction and tumor mediated immunosuppression. A clinical objective currently pursued is to use immunotherapy to ameliorate antitumour immunity. This will necessitate a high level immune response to ensure sufficient effector cells reach the tumor bed, focused cytotoxicity to eradicate malignant cells with little collateral damage to critical normal cells, and minimal inflammation. To achieve these aims, priority should be given to identifying more target antigens in astrocytoma and defining those cells present in the brain parenchyma that are essential to maintain antitumour effector function without exacerbating inflammation. If we are armed with better understanding of immune interactions with brain tumor cells, we can realistically envisage that immunotherapy will one day offer hope to patients with currently untreatable neoplastic diseases of the CNS. PMID- 12738054 TI - Ten years of antisense inhibition of brain G-protein-coupled receptor function. AB - Antisense oligonucleotides (AOs) are widely used as tools for inhibiting gene expression in the mammalian central nervous system. Successful gene suppression has been reported for different targets such as neurotransmitter receptors, neuropeptides, ion channels, trophic factors, cytokines, transporters, and others. This illustrates their potential for studying the expression and function of a wide range of proteins. AOs may even find therapeutic applications and provide an attractive strategy for intervention in diseases of the central nervous system (CNS). However, a lack of effectiveness and/or specificity could be a major drawback for research or clinical applications. Here we provide a critical overview of the literature from the past decade on AOs for the study of G-protein-coupled receptors (GPCRs). The following aspects will be considered: mechanisms by which AOs exert their effects, types of animal model system used, detection of antisense action, effects of AO design and delivery characteristics, non-antisense effects and toxicological properties, controls used in antisense studies to assess specificity, and our results (failures and successes). Although the start codon of the mRNA is the most popular region (46%) to target by AOs, targeting the coding region of GPCRs is almost as common (41%). Moreover, AOs directed to the coding region of the GPCR mRNA induce the highest reductions in receptor levels. To resist degradation by nucleases, the modified phosphorothioate AO (S-AO) is the most widely used and effective oligonucleotide. However, the end-capped phosphorothioate AOs (ECS-AOs) are increasingly used due to possible toxic and non-specific effects of the S-AO. Other parameters affecting the activity of a GPCR-targeting AO are the length (mostly an 18-, 20- or 21-mer) and the GC-content (mostly varying from 30 to 80%). Interestingly, one third of the AOs successfully targeting GPCRs possess a GC/AT ratio of 61-70%. AO induced reductions in GPCR expression levels and function range typically from 21 to 40% and 41 to 50%, respectively. In contrast to many antisense reviews, we therefore conclude that the functional activity of a GPCR after AO treatment correlates mostly with the density of the target receptors (maximum factor 2). However, AOs are no simple tools for experimental use in vivo. Despite successful results in GPCR research, no general guidelines exist for designing a GPCR targeting AO or, in general, for setting up a GPCR antisense experiment. It seems that the correct choice of a GPCR targeting AO can only be ascertained empirically. This disadvantage of antisense approaches results mostly from incomplete knowledge about the internalisation and mechanism of action of AOs. Together with non-specific effects of AOs and the difficulties of assessing target specificity, this makes the use of AOs a complex approach from which conclusions must be drawn with caution. Further antisense research has to be carried out to ensure the adequate use of AOs for studying GPCR function and to develop antisense as a valuable therapeutic modality. PMID- 12738055 TI - The insula (Island of Reil) and its role in auditory processing. Literature review. AB - The insular cortex is a complex structure which contains areas that subserve visceral sensory, motor, vestibular, and somatosensory functions. The role of the insular cortex in auditory processing was poorly understood until recently. However, recent case studies indicate that bilateral damage to the insulae may result in total auditory agnosia. Functional imaging studies demonstrate that the insulae participate in several key auditory processes, such as allocating auditory attention and tuning in to novel auditory stimuli, temporal processing, phonological processing and visual-auditory integration. These studies do not clarify the issue of further specialisation within the insular cortex, e.g. whether the posterior insulae are primarily sensory areas, while the anterior insulae serve mainly as integration/association auditory areas, two hypotheses that would be compatible with the cytoarchitectonic structure and connectivity of the insulae. The functional characterisation of the insulae remains incomplete, underlining the need for further studies. PMID- 12738056 TI - Methylenedioxymethamphetamine (MDMA, Ecstasy) neurotoxicity: cellular and molecular mechanisms. AB - Methylenedioxymethamphetamine (MDMA, Ecstasy) is a very popular drug of abuse. This has led to new intense concerns relevant to its nefarious neuropsychiatric effects. These adverse events might be related to the neurotoxic effects of the drug. Although the mechanisms of MDMA-induced neurotoxicity remain to be fully characterized, exposure to the drug can cause acute and long-term neurotoxic effects in animals and nonhuman primates. Recent studies have also documented possible toxic effects in the developing fetus. Nevertheless, there is still much debate concerning the effects of the drug in humans and how to best extrapolate animal and nonhuman primate data to the human condition. Herein, we review the evidence documenting the adverse effects of the drug in some animal models. We also discuss possible mechanisms for the development of MDMA neurotoxicity. Data supporting deleterious effects of this drug on the developing fetus are also described. Much remains to be done in order to clarify the molecular and biochemical pathways involved in the long-term neuroplastic changes associated with MDMA abuse. PMID- 12738057 TI - Calpain in the pathophysiology of spinal cord injury: neuroprotection with calpain inhibitors. AB - Spinal cord injury (SCI) evokes an increase in intracellular free Ca(2+) level resulting in activation of calpain, a Ca(2+)-dependent cysteine protease, which cleaves many cytoskeletal and myelin proteins. Calpain is widely expressed in the central nervous system (CNS) and regulated by calpastatin, an endogenous calpain specific inhibitor. Calpastatin degraded by overactivation of calpain after SCI may lose its regulatory efficiency. Evidence accumulated over the years indicates that uncontrolled calpain activity mediates the degradation of many cytoskeletal and membrane proteins in the course of neuronal death and contributes to the pathophysiology of SCI. Cleavage of the key cytoskeletal and membrane proteins by calpain is an irreversible process that perturbs the integrity and stability of CNS cells leading to cell death. Calpain in conjunction with caspases, most notably caspase-3, can cause apoptosis of the CNS cells following trauma. Aberrant Ca(2+) homeostasis following SCI inevitably activates calpain, which has been shown to play a crucial role in the pathophysiology of SCI. Therefore, calpain appears to be a potential therapeutic target in SCI. Substantial research effort has been focused upon the development of highly specific inhibitors of calpain and caspase-3 for therapeutic applications. Administration of cell permeable and specific inhibitors of calpain and caspase-3 in experimental animal models of SCI has provided significant neuroprotection, raising the hope that humans suffering from SCI may be treated with these inhibitors in the near future. PMID- 12738058 TI - Differential acute effects of fluoxetine on frontal and auditory cortex networks in vitro. AB - Primary cultures of neuronal networks grown on microelectrode arrays were used to quantify acute effects of fluoxetine (Prozac) on spontaneous spike and burst activity. For frontal cortex cultures, fluoxetine showed consistent inhibitory effects and terminated activity at 10-16 microM. IC(50) mean+/-S.E. for spike rates was 5.4+/-0.7 microM (n=15). For auditory cortex cultures, fluoxetine caused excitation at 1-10 microM, initial inhibition at 15 microM, and activity cessation at 20-25 microM. The spike rate IC(50) was 15.9+/-1.0 microM (n=11). Fluoxetine did not change the action potential waveform shape. However, at high concentrations, it caused total cessation of spike activity on all channels. The inhibition caused by fluoxetine was reversible for both tissues. Based on the results, we conclude that cultures showed repeatable, concentration-dependent sensitivities to fluoxetine but demonstrated tissue-specific responses for frontal and auditory cortex networks. These responses may not be due to the interference with serotonin reuptake, but may be due to a secondary effect on ionic channels. PMID- 12738060 TI - Neonatal cerebral hypoxia-ischemia causes lateralized memory impairments in the adult rat. AB - Neonatal hypoxia-ischemia (HI) has been extensively studied in a rat model characterized by unilateral brain damage (Rice-Vannucci Model). However, as well as in humans, each rat brain hemisphere is distinctly involved in cognitive functions, as for example retrieval of emotionally based memory, and neurochemical asymmetries have been described. In this paper we investigated whether hypoxia-ischemia could cause distinct cognitive deficits depending on which hemisphere is damaged. Seven-day-old male Wistar rats were submitted to permanent occlusion of left or right common carotid artery and were exposed to a mixture of 8% oxygen-92% nitrogen for 2.5 h. On adulthood, these rats were trained in step-down inhibitory avoidance and in two tasks in the Morris water maze. Both experimental groups (right and left lesioned) showed a deficit of retrieval in the inhibitory avoidance task compared to controls, although rats with right hemisphere lesion showed a significantly greater deficit than the left damaged group (P<0.05). In the Morris maze, both damaged groups presented cognitive deficits in the reference memory task (P<0.05), however only the right damaged group had an impairment in the working memory task. Brain coronal areas, at levels +1.20 and -3.30 mm from bregma of both HI groups were smaller than those of control, with no differences between the right and left damaged groups (P<0.05). These results show that cerebral hypoxia-ischemia in neonatal rats causes asymmetric behavioral outcomes depending on which of the hemispheres is lesioned and support the hypothesis of lateralization of cognitive functions in the rodent brain. PMID- 12738059 TI - Estrogen-induced autonomic effects are mediated by NMDA and GABAA receptors in the parabrachial nucleus. AB - The present study was done to determine if estrogen interacts with excitatory and/or inhibitory amino acid neurotransmitters to alter neuronal excitability within the parabrachial nucleus (PBN) and modulate autonomic tone. First, the role of estrogen in modulating autonomic tone was investigated in male rats anesthetized with Inactin (100 mg/kg). Animals were instrumented to record blood pressure, heart rate, vagal parasympathetic and renal sympathetic nerve activities as well as baroreflex sensitivity. Direct, bilateral injection of 17beta-estradiol (0.5 microM; 200 nl/side) into the PBN resulted in a significant decrease in blood pressure (17+/-4 mmHg), sympathetic tone (20+/-5%) and heart rate (22+/-5 beats/min) while increasing parasympathetic tone (34+/-4%) 30 min post-injection. These estrogen-induced effects were completely blocked by the co injection of estrogen with the estrogen receptor antagonist, ICI 182,780 (20 microM; 200 nl/side). Co-injection of the NMDA receptor antagonist, (+/-)-3-(2 carboxypiperazine-4-yl) propyl-1-phosphonic acid (CPP; 10 microM; 200 nl/side), with estradiol resulted in complete blockade of the estrogen-induced decrease in heart rate and increase in parasympathetic tone only. Co-injection of estradiol with the GABA(A) receptor antagonist, (+)-bicuculline (0.1 microM; 200 nl/side), resulted in complete blockade of the estrogen-induced decrease in blood pressure and sympathetic nerve activity only. These results suggest that estrogen acts on estrogen receptors on neurons in the PBN to modulate GABA(A)-receptor mediated inhibitory neurotransmission to alter sympathetic tone and blood pressure and on neurons in a separate, parallel pathway to modulate NMDA-receptor mediated neurotransmission to alter parasympathetic tone and heart rate. PMID- 12738061 TI - Identification of cerebral neurons and glial cell types inducing heat shock protein Hsp70 following heat stress in the rat. AB - Heat shock proteins were recently recognized as molecular chaperones that besides their chaperoning function were also involved in processes of cell death and survival. Many types of neural cells were reportedly capable of expressing heat shock protein Hsp70 following heat stress in vitro. However, identification of cell types inducing Hsp70 protein in the hyperthermic brain is not clear. In this study, cerebral Hsp70 distribution was evaluated in anesthetized adult rats (urethane, 1.5 g/kg, i.p.) subjected to short-term hyperthermia (41.5 degrees C for 30 min). Detection of Hsp70 was achieved by an ABC technique in vibratome or paraffin sections combined with specific markers of glial cell types. Hsp70 appeared by 90 min, mainly in glial and vascular cells, with enhanced immunostaining by 4 h following hyperthermia. Higher numbers of Hsp70-positive cells were detected in the white matter and diencephalic region than in the cerebral cortex, especially over the shorter interval. Hsp70 was localized in many oligodendrocytes, double-labeled with lectin GSII, and some vessels. Microglia showed apparently less Hsp70/OX-42 double-labeled cells than the previous two cell types. In contrast, only a few Hsp70-stained cells were positive for astrocyte marker GFAP. In addition to glial/vascular Hsp70 staining, neuronal Hsp70 induction was observed only in discrete regions including the paraventricular, supraoptic, suprachiasmatic and other hypothalamic nuclei, and in amygdala. Prevailing heat-stress expression of Hsp70 in oligodendrocytes and vascular cells might render them less susceptible to the consequences of other types of cell stress and could be exploited to increase selectively their survival in pathological situations. PMID- 12738062 TI - Dopamine effect on the stimulus pattern related changes in response characteristics of R/G horizontal cells in carp retina. AB - Repetitive red flashes increased the R/G horizontal cells' red response amplitude and induced a hyperpolarization of the cells' dark membrane potential. These phenomena were eliminated in 6-OHDA pretreated retinas and restored by exogenous dopamine, which suggests the involvement of dopamine receptor activity changes instead of dopamine release changes. Furthermore, the phenomena persisted on D(1) receptor antagonist (SKF-83566) application, whereas they diminished on D(2) receptor antagonist (eticlopride) application, indicating that the mechanism is related to a D(2) receptor, possibly located on photoreceptors. PMID- 12738063 TI - Analgesia and c-Fos expression in the periaqueductal gray induced by electroacupuncture at the Zusanli point in rats. AB - The need to use anaesthetised or restrained animals in acupuncture research in laboratory animals may represent a confounding variable, since both anaesthesia and stress alter the pain threshold and the activity of pain-related brain areas. In the current study we assessed the participation of the periaqueductal gray (PAG) in electroacupuncture's (EA) analgesic effects applied to the Zusanli point (36S) under carefully controlled stress conditions. Repeated immobilisation protocols (6 days, 1 h/day and 13 days, 2 h/day) were used to diminish the influence of acute immobilisation stress on c-Fos expression and analgesia (tail flick test) induced by electroacupuncture on the 36S point (EA36S). Animals submitted to immobilisation alone (IMMO) or to electroacupuncture (100 Hz, 2-4 V, faradic wave) on a non-point region (EANP) were compared with animals submitted to electroacupuncture on the 36S point. In animals not previously submitted to repeated immobilisation, electroacupuncture on the 36S point induced analgesia and c-Fos expression in the PAG was not different from that induced by electroacupuncture at a non-acupuncture point. In animals submitted to repeated immobilisation (repeated immobilisation for 6 days or repeated immobilisation for 13 days), however, electroacupuncture on point 36S led to higher levels of analgesia and c-Fos expression, specifically in the ventrolateral PAG (vlPAG), as compared with animal groups subjected only to immobilisation or to electroacupuncture on a non-point. Our findings endorse previous results, and point to a specific part of the PAG involved in the effects of electroacupuncture at the Zusanli point. PMID- 12738064 TI - Genetic differences in NMDA and D1 receptor levels, and operant responding for food and morphine in Lewis and Fischer 344 rats. AB - Previously, we have shown that Lewis (LEW) rats acquire faster than Fischer 344 (F344) rats operant food- and morphine-reinforced tasks under fixed-ratio schedules of reinforcement. The first purpose of the present work has been to study if differences in operant responding behavior may participate in the reported differences in morphine self-administration behavior between both inbred rat strains. To this end, we have analyzed the microstructure of responding obtained under a variable-interval (VI) of food reinforcement by calculating the inter-response time (IRT) for each rat strain. LEW rats exhibited shorter IRTs than F344 rats, suggesting that LEW rats may have an inherent high or compulsive operant responding activity. When subjects of both inbred rat strains were submitted to a schedule of morphine reinforcement of high responding requirements such as progressive ratio schedules, LEW rats also reached significantly higher breaking points and final response ratio than F344 rats for i.v. morphine self administration. Given that there are neurochemical differences between both rat strains and that glutamatergic N-methyl-D-aspartate (NMDA) and dopaminergic D(1) receptors have been involved in operant responding behavior, a second purpose of this work has been to measure basal NMDA and D(1) receptor levels in these rat strains by quantitative receptor autoradiography. Compared to F344 rats, LEW rats showed higher basal NMDA receptor levels in frontal and cingulate cortex, caudate putamen, central amygdaloid nuclei, and intermediate white layer of superior colliculus, and higher basal D(1) receptor levels in several areas of hippocampus and thalamus, and substantia nigra pars reticulata. Taken together, these results suggest that an inherent high operant responding activity of LEW rats may have a role in the previous reported faster acquisition of opiate-reinforced behavior in operant self-administration paradigms under fixed-ratio schedules of reinforcement. In addition, a basal higher NMDA and D(1) receptor levels of LEW rats compared to F344 rats may participate in the neurochemical background that mediates the behavioral differences between both inbred rat strains. PMID- 12738065 TI - Spontaneous sleep in mice with targeted disruptions of neuronal or inducible nitric oxide synthase genes. AB - Nitric oxide (NO) affects almost every physiological process, including the regulation of sleep. There is strong evidence that NO plays an important role in rapid eye movement sleep (REMS) regulation. To further investigate the role of NO in sleep, we characterized spontaneous sleep in mice with targeted disruptions (knockout; KO) in the neuronal nitric oxide synthase (nNOS) or inducible (i)NOS genes. REMS in nNOS KO mice was substantially lower than that of their control mice. In contrast, the iNOS KO mice had significantly more REMS than their controls. Inducible NOS KO mice also had less non-REMS (NREMS) during the dark period. Results suggest that nNOS and iNOS play opposite roles in REMS regulation. PMID- 12738066 TI - Distribution of corticotropin releasing hormone receptor immunoreactivity in the rat hypothalamus: coexpression in neuropeptide Y and dopamine neurons in the arcuate nucleus. AB - An abundance of physiological data suggests an interaction between neuropeptide Y (NPY) and corticotropin-releasing hormone (CRH) in the regulation of endocrine and autonomic functions. Previously, studies in our laboratory have indicated that NPY neurons in the arcuate nucleus of the hypothalamus (ARH) project to and come in close contact with CRH neurons in the paraventricular nucleus of the hypothalamus (PVH). Conversely, it has been demonstrated that the ventromedial portion of the ARH, an area containing NPY neurons, displays CRH receptor binding and CRH receptor mRNA. These data suggest a possible reciprocal feedback regulation between NPY and CRH neurons. The ARH also contains several other populations of neurons that may be targets of the CRH system and express CRH receptors; most notable are tuberoinfundibular dopaminergic neurons (TIDA). The PVH is an important component in the regulation of prolactin secretion and may play a role in the suppression of TIDA activity, which is a critical step in the prolactin stress response. The purpose of the present study was to characterize the distribution and cellular localization of CRH R(1) receptor-like immunoreactivity (CRH R(1)-ir) in the rat hypothalamus and to determine the phenotype of neurons in the ARH that contain CRH R(1)-ir. CRH R(1)-ir was present throughout the rat brain. Hypothalamic regions with the highest levels of immunostaining were the supraoptic nucleus, magnocellular PVH, ARH, and suprachiasmatic nucleus. Double label immunofluorescence was used to demonstrate that CRH R(1)-ir in the ARH was localized to NPY cell bodies. Furthermore, TIDA neurons in the ARH also displayed CRH R(1)-ir. However, despite an abundance of CRH R(1)-ir cells in the ARH, CRH-ir fiber innervation to the ARH was extremely sparse. Therefore, although this study provides neuroanatomical evidence for direct CRH R(1) regulation of ARH NPY and TIDA neurons in the rat, it is not consistent with the idea of a reciprocal feedback loop and suggests the involvement of other CRH-like ligands, such as urocortin. PMID- 12738067 TI - Effects of thyroid hormone on food intake, hypothalamic Na/K ATPase activity and ATP content. AB - The effects of thyroid hormone on whole body energy metabolism and compensatory effects on food intake are well established. However, the hypothalamic mechanisms that translate perceived whole body energy demands into subsequent appetitive behavior are incompletely understood. In order to address this question, we tested the effects of T3 on food intake and body weight in rats and measured neuronal Na/K ATPase activity and ATP content in the hypothalamus. Intraperitoneal T3 (100 microg/kg BW) administered for 6 consecutive days increased 24-h rat food intake from control, 26.6+/-1.2, to T3-treated 33.2+/-1.6 g (P<0.01). In T3-treated rats, rubidium-86 (86Rb) uptake (measured as a marker of Na/K ATPase activity) in ex vivo hypothalamic tissue increased (P<0.01) while the content of ATP in the ventral hypothalamus declined following T3 treatment (P<0.01). In another model of energy deficit, which was induced by a very low calorie diet, ATP content was also reduced in the hypothalamus compared to rats fed ad libitum. In summary, increased food intake in response to T3 may be secondary to decreased hypothalamic ATP content, perhaps resulting from both increased Na/K ATPase activity in the hypothalamus and metabolic signaling induced by whole body caloric deficit. PMID- 12738068 TI - Effects of isoflurane on auditory middle latency (MLRs) and steady-state (SSRs) responses recorded from the temporal cortex of the rat. AB - Auditory steady-state responses (SSRs) are believed to result from superimposition of middle latency responses (MLRs) evoked by individual stimuli during repetitive stimulation. Our previous studies showed that besides linear addition of MLRs, other phenomena, mainly related to the adaptive properties of neural sources, interact in a complex way to generate the SSRs recorded from the temporal cortex of awake rats. The aim of this study was to evaluate the effects of the inhalational general anesthetic, isoflurane, on MLRs and SSRs at several repetition rates (30-60 Hz) recorded from the temporal cortex of rats. Auditory evoked potentials were obtained by means of epidural electrodes in the awake condition and during anesthesia at three isoflurane concentrations (0.38, 0.76 and 1.13 vol.% in oxygen). MLR latency significantly increased during anesthesia in a concentration-dependent manner, while MLR amplitude, even when significantly attenuated with respect to the mean awake baseline value, failed to correlate with isoflurane concentration. SSRs decreased in amplitude and increased in phase during anesthesia in a concentration-dependent manner and the anesthetic-induced decrease of SSR amplitude appeared to be higher than the corresponding MLR attenuation. SSR prediction curves synthesized by linear addition of MLRs failed to predict SSRs in both amplitude and phase. Moreover, phase discrepancies proved to be higher during anesthesia. Our results suggest that MLRs and SSRs recorded from the temporal cortex of the rat exhibit differential sensitivity to isoflurane and that isoflurane could enhance the role of rate-dependent effects in SSR generation. PMID- 12738070 TI - Trigeminal modulation of gustatory neurons in the nucleus of the solitary tract. AB - Electrophysiological methods were used to investigate the effects of trigeminal nerve stimulation or transection on responses of single gustatory neurons in the nucleus of the solitary tract (NTS) to tastants (NaCl, sucrose, citric acid, monosodium glutamate) in pentobarbital-anesthetized rats. Unilateral transection of the lingual nerve, or the mandibular branch of the trigeminal nerve, resulted in significant reductions (by 21 and 29%, respectively; P<0.01) in tastant-evoked responses, with no further effect following bilateral transection. Electrical stimulation of the central cut end of the mandibular nerve directly excited nine of 14 gustatory NTS units. For these units, central mandibular stimulation facilitated the tastant-evoked responses in six, depressed responses in three, and had no effect in five. Facilitation of tastant-evoked responses peaked 4 min after mandibular stimulation and recovered within 8 min. Electrical stimulation of the peripheral cut end of the mandibular nerve significantly reduced tastant evoked responses in nine other NTS units, with a maximal reduction at 4 min post stimulation followed by recovery. Stimulation of the superior cervical sympathetic ganglion did not affect NTS tastant-evoked responses. These results suggest the presence of complex central modulation of NTS neurons by trigeminal afferents, as well as a peripheral depressant effect on gustatory processing possibly mediated via neuropeptide release from trigeminal nerve endings in the tongue. PMID- 12738069 TI - SYM 2081, an agonist that desensitizes kainate receptors, attenuates capsaicin and inflammatory hyperalgesia. AB - Excitatory amino acids acting at non-NMDA receptors contribute to transmission of nociceptive information. SYM 2081 ((2S,4R)-4-methyl glutamic acid) desensitizes kainate receptors, one subtype of non-NMDA receptors, to subsequent release of excitatory amino acids and thus may attenuate transmission of nociceptive information. To determine if SYM 2081 can prevent development of hyperalgesia, SYM 2081 (10, 50 or 100 mg/kg, i.p.) was administered prior to injection of capsaicin into the hindpaw of rats, which produces mechanical and heat hyperalgesia. To determine if SYM 2081 can reduce ongoing inflammatory hyperalgesia, SYM 2081 (10 or 100 mg/kg, i.p.) was administered after development of carrageenan-evoked hyperalgesia. Intraplantar injection of capsaicin produced an increase in hindpaw withdrawal frequency to mechanical stimuli (from 4+/-2 to 41+/-7%; mean+/-S.E.M.) and a decrease in withdrawal latency to heat (from 12.3+/ 0.3 to 5.9+/-0.4 s) in rats that received vehicle. In contrast, rats that received SYM 2081 (100 mg/kg) prior to injection of capsaicin exhibited a lower hindpaw withdrawal frequency (18+/-4%) and a longer withdrawal latency (7.7+/-0.5 s). Intrathecal (1-100 microg/5 microl), but not intraplantar (10 or 100 microg/50 microl), injection of SYM 2081 attenuated the development of capsaicin evoked heat hyperalgesia suggesting that SYM 2081's antihyperalgesic effects were due to its central effects. Furthermore, SYM 2081 completely reversed ongoing carrageenan-evoked mechanical hyperalgesia and partially (approximately 50%) reversed ongoing heat hyperalgesia. The present study demonstrates that administration of a high-potency ligand that selectively desensitizes kainate receptors attenuates the development of mechanical and heat hyperalgesia and attenuates ongoing inflammatory hyperalgesia. PMID- 12738071 TI - Sardinian alcohol-preferring and non-preferring rats show different reactivity to aversive stimuli and a similar response to a natural reward. AB - Sardinian alcohol-preferring (sP) and non-preferring (sNP) rats were studied to ascertain whether some behavioral and/or neurochemical traits, beyond ethanol preference, differentiated the two lines. Spontaneous reactivity of Wistar, sP and sNP rats to aversive or pleasurable stimuli was examined in an avoidance test, an elevated plus maze test, and in response to palatable food presentation. As the mesocorticolimbic dopaminergic system plays a relevant role in the response to rewarding or aversive stimuli, extraneuronal dopamine levels and cocaine-induced dopamine accumulation in the nucleus accumbens shell (NAcS) and medial prefrontal cortex (mPFC) were studied by microdialysis in the three groups of rats. Moreover, rats were exposed to repeated unavoidable stress and their avoidance response and NAcS dopamine output were determined. Finally, the capacity of sP, sNP, and Wistar rats to learn a palatable food-sustained appetitive behavior was studied. The present study shows that, beyond ethanol preference, there are several behavioral and neurochemical distinctions between sP and sNP rats. The sP rats displayed an increased level of anxiety-like behavior and sNP rats showed a reduced avoidance of noxious stimuli, compared to Wistar rats. Moreover, in the NAcS and PFC, extraneuronal dopamine levels were higher in sP rats and lower in sNP rats compared to Wistar rats; cocaine-induced dopamine accumulation in the NAcS was higher in sP rats than in sNP and Wistar rats. However, sP and sNP rats showed a similar behavioral and neurochemical response to chronic unavoidable stress. Interestingly, they also showed similar behavioral and neurochemical responses to a natural rewarding stimulus and a similar ability to learn an appetitive behavior. PMID- 12738072 TI - Comparative behavioral changes between male and female postpubertal rats following neonatal excitotoxic lesions of the ventral hippocampus. AB - Neonatal ventral hippocampal (nVH) lesioned male rat has been used as a model to test the hypothesis that early neurodevelopmental abnormalities lead to behavioral changes putatively linked to schizophrenia. There are significant gender differences in schizophrenia with male and female individuals differing in the age of onset, course and outcome of the disorder. In order to assess whether the behavioral effects of nVH lesions extend to or are different in female rats, we investigated spontaneous locomotion, grooming, social interactions and spatial memory in male and female rats post-pubertally at postnatal day (P) 56 following bilateral ibotenic acid of the ventral hippocampus at P7. The spontaneous locomotor activity in a novel environment of both male and female nVH lesioned rats was significantly enhanced compared to their respective sham-operated controls. In tests of social interactions, the number of encounters was significantly decreased in female lesioned rats, whereas the male nVH lesioned rats showed a significantly reduced duration of active social interactions. Furthermore, Morris water maze test showed a deficit of spatial learning/memory in only male lesioned rats with significant decrease in the latency to find hidden platform. These results suggest that while nVH lesions affect post pubertal behavior in both sexes of rats, the males appear to be affected to a greater extent than the females underscoring the influence of sex differences in the development of behaviors in the nVH lesioned animals. PMID- 12738073 TI - REM sleep deprivation-induced deficits in the latency-to-peak induction and maintenance of long-term potentiation within the CA1 region of the hippocampus. AB - Sleep loss adversely affects certain types of cognitive processing, particularly associative memory. Given that long-term potentiation (LTP) represents a putative cellular basis of learning and memory consolidation, the influence of sleep deprivation on LTP was examined. Rats were REM sleep deprived for 24, 48, or 72 h using the inverted flowerpot method in temperature-regulated chambers. Hippocampal slices taken from sleep-deprived rats were compared with home cage and pedestal control animals at 5, 15 and 60 min post-tetanization. The results indicated that at 5 min post-tetanization there were no differences in field potentials in any of the sleep-deprived or control groups, suggesting comparable levels of induction. However, analysis of latency-to-peak slope indicated that members of the 48 and 72 h sleep-deprived groups required approximately twice as long to achieve maximum slope as the 24 h group, home cage or 24, 48, 72 h pedestal controls (means 8.17, 7.50, 2.67, 4.67 and 3.17 min, respectively). At 15 min post-tetanization there were no group differences, however at 60 min post tetanization the slopes of the field excitatory postsynaptic potentials were significantly diminished for the 24, 48 and 72 h sleep-deprived groups (means 30.44, -1.89, 1.47, respectively) as compared with home cage and pedestal controls (means 59.54, 58.42, respectively). This delay in maximal induction, and the degradation of the maintenance phase of LTP, may represent sleep deprivation induced impairment of the underlying neurochemical mechanisms normally responsible for memory acquisition. PMID- 12738074 TI - Dextromethorphan prevents the diethyldithiocarbamate enhancement of 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine toxicity in mice. AB - In this report we show that dextromethorphan, a non-opioid cough suppressant, prevents the neurodegeneration of dopaminergic neurons in the substantia nigra of mice treated with diethyldithiocarbamate (DDC) and 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP). This effect is further substantiated by the assessment of dopamine (DA) content in the striatum of these animals. Dextromethorphan does not attenuate the striatal DA fall induced by MPTP alone but completely prevents DDC-induced enhancement after the combined treatment. Moreover, a study of DA metabolites has confirmed this neuroprotective property. The striatal levels of serotonin, which were studied as a control neuronal marker, did not change with any of the treatments administered. Furthermore, we show that dextromethorphan reduces the toxicity of glutamate against dopamine neurons in mesencephalic cell cultures. In line with previous data suggesting that dextromethorphan can prevent neuronal damage, our observations supply new evidence regarding the possibility of this compound being of therapeutic use in neurodegenerative diseases. PMID- 12738075 TI - The GABAA receptor agonist THIP is neuroprotective in organotypic hippocampal slice cultures. AB - The potential neuroprotective effects of the GABA(A) receptor agonists THIP (4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol) and muscimol, and the selective GluR5 kainate receptor agonist ATPA ((RS)-2-amino-3-(3-hydroxy-5-tert butylisoxazol-4-yl)propanoic acid), which activates GABAergic interneurons, were examined in hippocampal slice cultures exposed to N-methyl-D-aspartate (NMDA). The NMDA-induced excitotoxicity was quantified by densitometric measurements of propidium iodide (PI) uptake. THIP (100-1000 microM) was neuroprotective in slice cultures co-exposed to NMDA (10 microM) for 48 h, while muscimol (100-1000 microM) and ATPA (1-3 microM) were without effect. The results demonstrate that direct GABA(A) agonism can mediate neuroprotection in the hippocampus in vitro as previously suggested in vivo. PMID- 12738076 TI - Effects of Solanum melongena (garden egg) on some visual functions of visually active Igbos of Nigeria. AB - Solanum melongena LINN (garden egg) or bitter tomato is an economic flowering plant belonging to the family Solanaceae and widely distributed throughout the temperate and tropical regions. Members are mostly herbaceous plants, and the fruit is berry and the seeds have large endosperm and are grown mainly for food and medicinal purposes. The effects of bolus consumption of 10 g of S. melongena were undertaken on visually active male volunteers so as to determine its ocular complications. Results showed that the pupil size was reduced (23%), NPC was decreased (9%) and the AA was increased (22%) and the intraocular pressure dropped by 25% while there was no effect on VA, and the habitual phoria. The miotic effect lowered the intraocular pressure appreciably and the reduced NPC which was still within normal range did not produce any vision discomfort. The increased AA and convergence excess positively correlated provide an efficient visual mechanism. It is suggested that S. melongena would be of benefit to patients suffering from raised intraocular pressure (glaucoma) and convergence insufficiency. PMID- 12738077 TI - Ocular dynamics of systemic aqueous extracts of Xylopia aethiopica (African guinea pepper) seeds on visually active volunteers. AB - Xylopia aethiopica, African guinea pepper, is an angiosperm belonging to the family Annonecae, and used mainly as spice and in traditional medicine. The ocular dynamics of bolus consumption of 300 mg total dose was undertaken on visually active volunteers with a view to finding its ocular effects or complications. Results showed that the aqueous extract of X. aethiopica was neither a miotic nor a mydriatic, but lowered the intraocular pressure (17.48%), reduced the near point of convergence (31.1%) and increased the amplitude of accommodation (8.98%) which are positively correlated (r=0.95). On the other hand, the systemic extract had no effect on the visual acuity at far and near as well as the phoria status at the appropriate distances. The convergence excess resulted in esophoria and the increased amplitude of accommodation placed greater demand on accommodation mechanism without any discomfort. The nonspecific mechanism of action makes it a safer spice which can be exploited in the management of exophoria and raised intraocular pressure (glaucoma) in instances where the efficacy of the older conventional drugs is insufficient. PMID- 12738078 TI - Ethnobotanical survey and in vitro antiplasmodial activity of plants used in traditional medicine in Burkina Faso. AB - In Burkina Faso, most people in particular, in rural areas, use traditional medicine and medicinal plants to treat usual diseases. In the course of new antimalarial compounds, an ethnobotanical survey has been conducted in different regions. Seven plants, often cited by traditional practitioners and not chemically investigated, have been selected for an antiplasmodial screening: Pavetta crassipes (K. Schum), Acanthospermum hispidum (DC), Terminalia macroptera (Guill. et Perr), Cassia siamea (Lam), Ficus sycomorus (L), Fadogia agrestis (Schweinf. Ex Hiern) and Crossopteryx febrifuga (AFZ. Ex G. Don) Benth. Basic, chloroform, methanol, water-methanol and aqueous crude extracts have been prepared and tested on Plasmodium falciparum chloroquine-resistant W2 strain. A significant activity has been observed with alkaloid extract of P. crassipes (IC(50)<4 microg/ml), of A. hispidum, C. febrifuga, and F. agrestis (4140 mmHg) received HS (250 mg/kg/day) in drinking water (2K-1C+HS). The second group (2K 1C) and the sham-operated (Sh-Op) controls, received drinking water. BP was monitored weekly using rat-tail plethysmography. After 8 weeks, 2K-1C+HS had a reduction in systolic BP (139.6+/-1.6 mmHg) compared to 2K-1C (174+/-2.4 mmHg, n=5; P<0.001). No significant difference was found in BP of 2K-1C+HS and Sh-Op (139.6+/-1.6 mmHg versus 132+/-3.4 mmHg). A reduction in heart rate in 2K-1C+HS was observed (388+/-3.7 bpm versus 444+/-6.8 bpm in 2K-1C and 416+/-9.3 in Sh-Op, n=5; P<0.001). The hearts of 2K-1C were heavier than those of 2K-1C+HS (0.74+/ 0.03 g versus 0.66+/-0.03 g, n=5; P<0.05). Cardiac weight of 2K-1C+HS was comparable to those of Sh-Op (0.57+/-0.04 g). Serum creatinine and plasma electrolytes were not different from controls. This study suggests that HS exhibits antihypertensive and cardioprotective effects in vivo and supports the public belief that HS may be a useful antihypertensive agent. PMID- 12738085 TI - Antispasmodic effect of three fractions of hydroalcoholic extract of Pycnocycla spinosa. AB - Hydroalcoholic extract of Pycnocycla spinosa was shown to have spasmolytic action in vitro and antidiarroheal effect in vivo. The hydroalcoholic extract of P. spinosa is composed of alkaloid-, flavonoid- and saponin-rich components. These components were separated by fractional separation technique. The pharmacological objective of this study was to look for relaxant effect of these components of hydroalcoholic extract of P. spinosa on rat isolated ileum contractions. The alkaloid-, flavonoid- and saponin-rich fractions of P. spinosa inhibited the response to 80 mM KCl in a concentration-dependent manner. The alkaloid-rich fraction was the most and saponin-rich fraction the least effective relaxant of ileum contractions. The alkaloid-rich fraction also inhibited the response to acetylcholine (ACh) and 5-hydroxytryptamine (5-HT). From this study, it can be concluded that the antispasmodic action of hydroalcoholic extract of P. spinosa could be due to components in the alkaloid-rich fraction and to a lesser extent, to its flavonoid-rich fraction. PMID- 12738086 TI - Effect of fruits of Moringa oleifera on the lipid profile of normal and hypercholesterolaemic rabbits. AB - Rabbits were fed Moringa oleifera (200mg/kg/day, p.o.) or lovastatin (6mg/kg/day, p.o.) in banana pulp along with standard laboratory diet and hypercholesterolaemic diet for 120 days. Moringa oleifera and lovastatin were found to lower the serum cholesterol, phospholipid, triglyceride, VLDL, LDL, cholesterol to phospholipid ratio and atherogenic index, but were found to increase the HDL ratio (HDL/HDL-total cholesterol) as compared to the corresponding control groups. Treatment with M. oleifera or lovastatin in normal rabbits decreased the HDL levels. However, HDL levels were significantly increased or decreased in M. oleifera- or lovastatin-treated hypercholesterolaemic rabbits, respectively. Lovastatin- or M. oleifera-treated hypercholesterolaemic rabbits showed decrease in lipid profile of liver, heart and aorta while similar treatment of normal animals did not produce significant reduction in heart. Moringa oleifera was found to increase the excretion of faecal cholesterol. Thus, the study demonstrates that M. oleifera possesses a hypolipidaemic effect. PMID- 12738087 TI - Hepatoprotective activity of leaves of Kalanchoe pinnata Pers. AB - Kalanchoe pinnata Pers. is naturalized throughout the hot and moist parts of India. Juice of the fresh leaves is used very effectively for the treatment of jaundice in folk medicines of Bundelkhand region of India. The juice of the leaves and the ethanolic extract of the marc left after expressing the juice were studied in rats against CCl(4)-induced hepatotoxicity. The test material was found effective as hepatoprotective as evidenced by in vitro, in vivo and histopathological studies. The juice was found more effective than ethanolic extract. PMID- 12738088 TI - Efficacy of different Cynara scolymus preparations on liver complaints. AB - Cynara scolymus leaves extracts have long been used in folk medicine for their choleretic and hepatoprotective activities, that are often related to the cynarin content. These therapeutic properties are also attributed to mono- and di caffeoylquinic acids and since commercial C. scolymus preparations can differ for their activities, we studied four extracts to evaluate, if present, a relationship between the hepatobiliary properties of the different preparations and their content in phenolics. The antioxidant activity of the commercial preparations examined was also considered in an in vitro system. The results showed that the extract with the highest content in phenolic derivatives (GAE) exerted the major effect on bile flow and liver protection. Also the results of the antioxidant capacity (BR) of the different preparations are in good agreement with the results obtained in vivo. On the contrary, administering rats with doses of chlorogenic acid, equivalent to those present in this extract, we did not observe any choleretic or protective action. An histopathological analysis of liver sections confirmed the biochemical results. Perhaps caffeoyl derivatives have a role in the therapeutic properties of C. scolymus extracts, as reported in literature for "in vitro" studies, but when administered alone, they are not so effective in exerting this action. PMID- 12738090 TI - Investigation of antihypertensive mechanism of garlic in 2K1C hypertensive rat. AB - This study sought to examine the antihypertensive mechanism of garlic in two kidney-one-clip (2K1C) hypertensive rat. In this study, the effect of garlic on serum and tissue including: aorta, heart, kidney, lung as well as circulatory (serum) ACE activity in 2K1C rats were examined. Four groups of rats were selected: control "CTL", sham-operated "SHAM", hypertensive "H" and garlic treated hypertensive "GT" group. Hypertension was induced by surgery. Four weeks post-clipping, single daily dose of 50mg of aqueous extract of garlic was given orally to "GT" rats for 4 weeks. Blood pressure was measured by tail-cuff method. ACE activity was determined using HPLC. The systolic blood pressure (SBP) was significantly increased in "H" compared to "CTL" group. In "GT" group, blood pressure was significantly decreased compared to "H" group. The ACE activity in all tissues of "H" group was significantly increased compared to controls which was significantly decreased in garlic-treated compared to non-treated hypertensive rats. These results indicated a negative correlation between consumption of garlic, blood pressure and ACE activity in serum and different tissues in 2K1C rats, suggesting that garlic has a significant blood pressure lowering effect, which could partly be mediated by reduction in ACE activity. PMID- 12738089 TI - Endothelium-dependent effects of the ethanolic extract of the mistletoe Psittacanthus calyculatus on the vasomotor responses of rat aortic rings. AB - The mistletoe Psittacanthus calyculatus (Loranthaceae) is used in Mexican traditional medicine for the treatment of hypertension. In the present study the effects of a crude ethanolic extract of this mistletoe, on the vasomotor reactivity of superfused rat aortic rings (with or without a functional endothelium) were analyzed. Either in the absence or in the presence of L-NAME or indomethacin, the extract (12.5-800 microg/ml) had no effect on the basal tone of both types of rings. In phenylephrine-precontracted rings, low concentrations of the extract (up to 300 microg/ml) induced a small additional tension development in both types of rings; however, the tension increase was slightly larger in rings having an intact endothelium. At higher concentrations (400-800 microg/ml), the extract relaxed, concentration-dependently, phenylephrine-precontracted rings with an intact endothelium. This relaxation was completely reverted by the addition of L-NAME. When the extract was applied in the continuous presence of L NAME to phenylephrine-precontracted rings, instead of a relaxation a marked additional tension development occurred. Indomethacin did not modify the relaxation induced by the extract. The results indicate that the ethanolic extract of this mistletoe induces, predominantly, an endothelium-dependent relaxation which seems to be mediated by the synthesis/release of nitric oxide. PMID- 12738091 TI - The effect of khat chewing on gallbladder motility in a group of volunteers. AB - An increase in the prevalence of gallstones has been reported from the Middle East and the Republic of Yemen. Changing dietary habits and obesity are thought to be responsible but other local factors may contribute such as chewing the leaves of the khat plant (Catha edulis Forsk.) which is a widespread social custom in Yemen. We have studied the effects of khat chewing on gallbladder motility in a group of 10 healthy volunteers. All subjects underwent ultrasound measurements of gallbladder volume after chewing khat leaves or lettuce, which was used as the control. Results were compared after chewing for up to 2h in the fasting state and in response to a fatty meal. There was no significant change in gallbladder volume after chewing khat compared with lettuce in the fasting state (P=0.7) or in gallbladder emptying after a fatty meal (P=0.4) and we conclude that khat chewing has no clinically significant effect on gallbladder motility. PMID- 12738092 TI - Analgesic, antipyretic, anti-inflammatory effects of methanol, chloroform and ether extracts of Vernonia cinerea less leaf. AB - The chloroform, methanolic and ether extracts of Vernonia cinerea (Asteraceae; Less) leaf (100, 200 and 400mg/kg intraperitoneally) were tested in: acetic acid induced writhing in mice, carrageenin-induced oedema and brewer's yeast-induced pyrexia in rats to assess their analgesic, anti-inflammatory, antipyretic and behavioral activities, respectively. The changes in writhings and behavioural activities in mice, the pyrexia and paw volumes in rats were reduced significantly (P<0.05) compared to the control. There was an increase in pain threshold on the oedematous right hind limb paw of the rats. These results indicate that the extracts could possess analgesic, antipyretic and anti inflammatory properties. All these effects and the changes in the behavioural activities could be suggested as contributory effects to the use of V. cinerea leaf in the treatment of malaria. PMID- 12738093 TI - Screening of traditionally used South African plants for antifungal activity against Candida albicans. AB - Twenty-four South African medicinal plants were screened against Candida albicans standard strain ATCC 10231 and two clinical isolates from a 5-month-old baby and an adult, in an attempt to find a traditional remedy to treat oral candidiasis, which is prevalent in HIV-patients. Allium sativum L. and Tulbaghia violacea L. aqueous bulb extracts had MIC values of 0.56 and 3.25mg/ml respectively, whilst Polygala myrtifolia L. leaves and Glycyrrhiza glabra L. rhizome extracts had MIC values of 1.56 mg/ml when tested against the 5-month-old isolate. Fresh water extracts stored at 4, 23 and 33 degrees C over a period of a week, were used to determine the stability of these extracts. Allium sativum and Tulbaghia violacea maintained activity at 4 degrees C, but not at higher temperatures, whereas Polygala myrtifolia and Glycyrrhiza glabra lost activity within a day even at 4 degrees C. The unpleasant taste of the two species with a garlic smell, could however not be masked, and as the smell following the eating of the two species would lead to HIV-patients being recognised, these two plants where not considered for further investigation. Therefore, Polygala myrtifolia and Glycyrrhiza glabra are being further investigated for use as an oral mouthwash in clinics and homes. PMID- 12738094 TI - Variation of leishmanicidal activity in four populations of Urechites andrieuxii. AB - Urechites andrieuxii Muell.-Arg. (Apocynaceae) is widely used in the Yucatan Peninsula for the treatment of cutaneous leishmaniasis. The influence of the environment in the variability of the leishmanicidal activity of the plant was evaluated using crude methanol extracts of roots from individuals belonging to four natural populations growing in the Yucatan Peninsula. The results of the growth inhibition test using three Leishmania spp. promastigotes showed a stronger leishmanicidal activity in populations of U. andrieuxii growing in more humid environments. Further evaluation against four human cancer cell lines and in the brine shrimp bioassay of both extracts from various parts of the plant and from the most active methanol root extracts, suggested that while the leaf extract appears to have selective toxicity against Leishmania parasites, the strong leishmanicidal activity detected in the root extracts of the plant might be due to its cytotoxicity. PMID- 12738095 TI - In vitro antiplasmodial activity of antimalarial medicinal plants used in Vietnamese traditional medicine. AB - Among 42 extracts, prepared from 14 medicinal plants used in Vietnamese traditional medicine to treat malaria, 24 were found to have antiplasmodial activity by inhibiting the growth of the chloroquine-resistant Plasmodium falciparum strain FCR-3 with EC(50) values less than 10 microg/ml. Each medicinal plant possessed at least one active extract. The methanol extract of Coscinium fenestratum had the strongest antiplasmodial activity with EC(50) value of 0.5 microg/ml. Activity-guided fractionation led to identification of berberine as the major active constituent. PMID- 12738096 TI - Anti-metastatic effects of aqueous extract of Helixanthera parasitica. AB - Metastasis, the spread of cancer in body, is a major cause of death. We have screened anti-metastatic activity of aqueous and dichloromethane extracts of several not previously studied Thai herbs, using an in vitro invasion test. This involves the in vitro invasion of HCC-S102, a hepatocellular carcinoma cell line derived from a Thai patient, through a reconstituted-basement membrane (Matrigel). The aqueous extract of a plant (Helixanthera parasitica) revealed a significant inhibitory effect on the cancer cell invasion, and showed antioxidant activity. The aqueous extract was partially purified by silica gel column chromatography, and the highest anti-metastatic activity fraction showed 83% inhibition of invasion with low cytotoxic effect. However, anti-metastatic activity was not associated with the antioxidant activity of the aqueous extract. PMID- 12738097 TI - Female genital cutting: a need for reform. PMID- 12738098 TI - Neuroma of the clitoris after female genital cutting. AB - BACKGROUND: Nerve tumors of the clitoris and particularly neuromas are extremely rare. CASE: A 27-year-old infibulated African woman suffering from chronic vulvar pain increasing with sexual intercourse presented for gynecologic care. Examination revealed a painful clitoral tumor. The tumor was surgically excised. The diagnosis of amputation neuroma of the clitoris was made by microscopic examination. CONCLUSION: This is the first well-documented case of clitoral amputation neuroma occurring after female genital cutting. Considering the high number of genital cuttings practiced, these tumors are probably under-reported in the literature. PMID- 12738099 TI - Urethral erosion of a tension-free vaginal tape. AB - BACKGROUND: Urethral dilation has been recommended to treat voiding dysfunction that may occur after placement of tension-free vaginal tape (TVT) for the treatment of stress urinary incontinence. We report on a case of urethral erosion by the tape secondary to repetitive urethral dilations after surgery. CASE: A urethral erosion by the tape was diagnosed by cystoscopy after three urethral dilations failed to resolve postoperative voiding dysfunction 8 weeks after the initial procedure. A partial tape revision with repair of the urethrotomy was performed, with resolution of the subject's voiding dysfunction. CONCLUSION: This report describes a potential complication of a recommended treatment for voiding dysfunction after placement of TVT. PMID- 12738100 TI - Vaginal evisceration long after vaginal hysterectomy. AB - BACKGROUND: Vaginal evisceration can take place many years after vaginal surgery. CASE: An 87-year-old woman presented with evisceration of small bowel through the vagina, 15 years after she underwent a vaginal hysterectomy. On physical examination, her vital signs were normal. Forty centimeters of small bowel was visible emerging from the vagina, appearing viable and nonedematous. Because of the high surgical risk, the bowel was replaced and the defect in the vaginal wall was repaired transvaginally. CONCLUSION: Vaginal evisceration can be treated by a transvaginal surgical approach. Factors such as the medical condition of the patient and the viability of the herniated viscus should dictate the optimal approach in each case. PMID- 12738101 TI - Metastatic unknown primary tumor presenting in pregnancy as multiple cerebral infarcts. AB - BACKGROUND: Cancer presenting during pregnancy is a rare event. There are no reports of an unknown primary tumor presenting during pregnancy. CASE: A 35-year old primigravida presented at 16 weeks' gestation with multiple cerebral infarcts. After a negative workup she was discharged on anticoagulation therapy, only to return at 29 weeks with diffuse, metastatic cancer of unknown primary origin. After an elective 32-week delivery she received aggressive chemotherapy but ultimately died 5 months later. CONCLUSION: Malignancy should be included in the differential diagnosis of unusual, unexplained cases of thromboembolism in pregnancy. PMID- 12738102 TI - Evidence of fetal C-reactive protein urinary excretion in early gestation. AB - BACKGROUND: C-reactive protein has been measured in amniotic fluid in the second and third trimesters of gestation, and its elevated concentration has been found to be associated with adverse pregnancy outcome. It remains unexplained whether amniotic fluid C-reactive protein is of fetal origin. CASE: We report the measurement of C-reactive protein in fetal urine obtained by transabdominal vescicocentesis in a fetus at 15 weeks' gestation affected by obstructive uropathy. Using an enzyme-linked immunosorbent assay, C-reactive protein was detected at a concentration of 234 ng/mL. CONCLUSION: The fetal kidneys excrete C reactive protein as early as 15 weeks' gestation. PMID- 12738103 TI - Weighted speculum buttock burn during gynecologic surgery. AB - Genital tract and perineal electrical burns during resectoscopic surgery are occasionally encountered. However, perineal or buttock burns from other causes have not been reported.A 44-year-old overweight woman with a history of menometrorrhagia underwent endometrial ablation with a thermal balloon device uneventfully under general anesthesia. Upon removal of the weighted speculum it was noted that the buttocks had sustained two oval burns under the speculum ball. Experiments demonstrated that the weighted speculum cools off differentially, with the ball temperature remaining at higher than 45C in room air for at least 30 minutes after autoclaving.The use of the weighted speculum within 30 minutes of autoclaving may result in buttock burns. The speculum cools to lower than 40C within 1 minute when rinsed or bathed with at least 1 L of saline solution. PMID- 12738104 TI - Vaginal hyperpigmentation due to ochronosis. AB - BACKGROUND: Ochronosis is a manifestation of alkaptonuria, a rare metabolic disorder. It results in the accumulation of pigment in connective tissues. After several years, ochronosis may produce a distinctive form of degenerative arthritis. CASE: Vaginal ochronosis was diagnosed in an asymptomatic elderly woman with vaginal hyperpigmentation and severe degenerative arthritis. CONCLUSION: Vaginal hyperpigmentation is a rare clinical finding that necessitates biopsy. PMID- 12738105 TI - Prioritizing posterior arm delivery during severe shoulder dystocia. AB - BACKGROUND: Delivery of the posterior arm, or the Barnum maneuver, is at times used late in shoulder dystocia management algorithms, and is not often a first- or second-line management protocol. CASE: A multiparous, diabetic patient, who was morbidly obese and had a residual obstetric brachial plexus injury, experienced a precipitous second stage of labor and severe shoulder dystocia. Attempts at the McRoberts maneuver with traction failed to deliver the fetus. In lieu of alternative maneuvers or continued attempts at traction, the posterior arm was delivered and the fetal trunk followed easily. CONCLUSION: A geometric analysis reveals that using posterior arm delivery reduces the obstruction by more than a factor of two, relative to the McRoberts maneuver. We recommend earlier use of this maneuver during shoulder dystocia management. PMID- 12738106 TI - Reversible azoospermia: anabolic steroids may profoundly affect human immunodeficiency virus-seropositive men undergoing assisted reproduction. AB - BACKGROUND: In vitro fertilization (IVF) with intracytoplasmic sperm injection (ICSI) has recently been offered to human immunodeficiency virus (HIV) serodiscordant couples where the man is seropositive and the woman seronegative to achieve pregnancy while minimizing the risk of HIV transmission. Anabolic steroids are commonly prescribed medications for adjunctive treatment of HIV disease to prevent muscle wasting. CASE: An HIV-serodiscordant couple presented for fertility care and evaluation. The man was found to be azoospermic. Further evaluation attributed his azoospermia to his treatment with testosterone and oxandrolone. After these agents were discontinued, his azoospermia resolved within 3 months. Normal sperm were then cryopreserved for future use, and his medications were resumed. Later the couple conceived by IVF-ICSI using the cryopreserved sperm. CONCLUSION: The popular use of anabolic steroids in HIV infected men may predispose them to abnormal sperm production. PMID- 12738107 TI - Short delay of delivery to allow corticosteroid administration in a case of preterm antepartum eclampsia. AB - BACKGROUND: The current recommendation for management of antepartum eclampsia is to take steps to deliver the fetus after stabilization of the maternal condition. We delayed delivery for 60 hours in a case of antepartum preterm eclampsia to allow administration of corticosteroids for fetal lung maturity enhancement. CASE: A primigravida presented with eclamptic seizure at 29 weeks' gestation without focal neurological deficits. Clinical and laboratory assessment ruled out the presence of the syndrome of hemolysis, elevated liver enzymes, low platelets; abruption; disseminated intravascular coagulation; or acute renal failure. The fetal biometry was appropriate for gestational age, and there was a normal amount of amniotic fluid. Fetal testing was reassuring. Expectancy with intravenous magnesium sulfate infusion was continued for 60 hours, allowing administration of a course of corticosteroids for enhancement of fetal lung maturity. Maternal and neonatal outcomes were satisfactory. CONCLUSION: A 2-day delay in delivery in selected patients with preterm antepartum eclampsia allows administration of steroids for fetal lung maturity enhancement. PMID- 12738108 TI - An unusual case of ovarian resistance syndrome. AB - BACKGROUND: Women who present with elevated gonadotropins, primary amenorrhea, and sexual infantilism usually have an abnormal karyotype or premature ovarian failure. We describe a rare case of ovarian resistance secondary to a gonadotropin post-receptor defect. CASE: An 18-year-old presented with amenorrhea and sexual infantilism. Original workup led physicians to believe the patient had uterine agenesis with premature ovarian failure. Ovarian biopsy proved the presence of follicles. After 48 months on hormone replacement, the patient went through the stages of puberty, including menarche. Re-evaluation proved the presence of a uterus. CONCLUSION: Ovarian resistance is a rare cause of hypergonadotropic hypogonadism. With this type of ovarian dysfunction, women present early in life with amenorrhea, elevated gonadotropins, and normal karyotypes. PMID- 12738109 TI - Increased intracranial blood flow volume in a preeclamptic woman with postpartum photophobia. AB - BACKGROUND: Cerebral circulatory changes in preeclampsia are unclear. We studied the changes in intracranial blood flow volume using a new color Doppler ultrasonographic assessment in a preeclamptic woman with photophobia. CASE: A 39 year-old preeclamptic primigravida was admitted and delivered by cesarean at 36 weeks' gestation. She developed bilateral photophobia with blood pressure elevation at 2 days postpartum. Blood flow volume index [mean velocity x pid(2)/4] (d = luminal diameter at systolic phase) was established. The sum of blood flow volume indexes of the bilateral internal carotid arteries and vertebral arteries increased at the onset of photophobia and blood pressure elevation. The blood flow volume index increased above 120 mm Hg of mean arterial blood pressure. CONCLUSION: These data represent the increased cerebral hemodynamic changes in preeclampsia with photophobia. PMID- 12738110 TI - Prenatal sonographic findings of congenital duplication of the cecum. AB - BACKGROUND: Alimentary tract duplications are rare congenital lesions that may arise at any location throughout the gastrointestinal tract. The most common site of enteric duplications is the ileum, with only 13% of cases occurring in the colon. CASE: Prenatal sonography at 39 weeks' gestation showed a cystic structure with thick walls exhibiting clear peristaltic movements in the right lower fetal abdomen. Inner walls of the cystic structure had small folds suggestive of colonic haustra. The sonographic findings and location of the finding were consistent with duplication of the colon and, possibly, the cecum. Congenital duplication of the cecum was proven at neonatal laparotomy. Ileocecal resection with ileocolonic anatomosis was performed and congenital duplication of the cecum confirmed by pathology examination of the resected specimen. CONCLUSION: Unusual sites of alimentary tract duplications detected by prenatal ultrasound include the cecum. PMID- 12738111 TI - Giant uterine tumors: two cases with different clinical presentations. AB - BACKGROUND: Giant uterine tumors are uncommon. However, they may be life threatening because of pressure effects on the lungs and other adjacent organs. Proper surgical management and careful perioperative care are essential to assure a good outcome after excision. CASE: Two women with giant uterine leiomyomata (weighing more then 40 kg [88 lb]) are discussed. In one case the leiomyoma led to severe pulmonary hypertension and respiratory failure necessitating an emergency operation. Abdominal hysterectomy and bilateral salpingo-oophorectomy were successfully carried out in both cases, which are among the largest tumors ever removed with survival of the patient. CONCLUSION: Different clinical manifestations can be expected in cases of giant uterine tumors according to which other organs are secondarily affected. Numerous difficulties may be encountered in the evaluation and removal of these tumors. A combined team consisting of gynecologic, general, and plastic surgeons is necessary for a successful outcome. PMID- 12738112 TI - Hypodysfibrinogenemia during pregnancy, labor, and delivery. AB - BACKGROUND: Hypodysfibrinogenemia is an autosomally dominant disorder that can result in excessive bleeding as well as specific pregnancy complications. Increased risks of spontaneous abortion, postpartum hemorrhage, poor wound healing, and placental abruption have been reported. CASE: A woman with hypodysfibrinogenemia presented for care in the first trimester. Her antepartum course was uncomplicated, and she was administered intermediate-purity factor VIII during labor and did not have excessive bleeding postpartum. The infant's cord fibrinogen was low, at 43 mg/dL (normal 215 +/- 30), showing it was similarly affected. CONCLUSION: Pregnancy in patients with hypodysfibrinogenemia can be associated with various complications; however, coordination of care and anticipation of specific problems can result in a successful outcome for both mother and infant. PMID- 12738113 TI - Drug rash with eosinophilia and systemic symptoms syndrome and renal toxicity with a nevirapine-containing regimen in a pregnant patient with human immunodeficiency virus. AB - BACKGROUND: Antiretroviral therapy is associated with serious adverse events. We report the case of a pregnant human immunodeficiency virus type 1-infected woman who developed drug rash with eosinophilia and systemic symptoms syndrome and renal failure shortly after initiation of a nevirapine-containing antiretroviral regimen at 27 weeks' gestation. CASE: A 26-year-old primigravida presented with a fever of 40.2C, urticarial rash, and icteric sclera 6 weeks after starting a nevirapine-containing antiretroviral regimen. Eosinophils, serum creatinine, bilirubin, and liver enzymes were markedly elevated, and abnormal coagulation studies were noted on admission. Serology testing was negative for viral hepatitis and microbiologic cultures were negative for growth. Abnormal laboratory findings at discharge resolved within 4 months after discontinuation of antiretroviral agents and systemic corticosteroid therapy. CONCLUSION: Our case suggests the need for close monitoring of liver and renal function after initiation of nevirapine-containing antiretroviral regimens. PMID- 12738114 TI - Familial Mediterranean fever presenting as recurrent acute pelvic inflammatory disease. AB - BACKGROUND: Recurrent acute episodes of pelvic inflammatory disease (PID) often present a diagnostic dilemma. The differential diagnosis should include reinfection, appendicitis, endometriosis, irritable bowel syndrome, colitis, persistent ovarian cyst, and antibiotic-resistant bacterial strains. CASE: NA young Palestinian woman presented with recurrent episodes of pelvic pain with rebound tenderness, fever, and elevated white blood cell count, erythrocyte sedimentation rate, and C-reactive protein. The patient underwent extensive workup, multiple courses of intravenous and oral antibiotics, and diagnostic laparoscopies, with continued recurrent episodes. Treatment with colchicine for suspected familial Mediterranean fever resulted in resolution of symptoms. CONCLUSION: In patients of Mediterranean ancestry who have symptoms of recurrent PID that are refractory to conventional treatment, familial Mediterranean fever should be included in the differential diagnosis. PMID- 12738115 TI - Wandering spleen presenting as acute pancreatitis in pregnancy. AB - BACKGROUND: Wandering spleen most frequently presents clinically with gastrointestinal bleeding and thrombocytopenia. CASE: A 24-year-old multiparous patient at 36 weeks' gestation presented with pancreatitis. Incidental thrombocytopenia was discovered. A 2722-g female neonate was delivered by repeat cesarean from a breech presentation. The spleen was observed at the left margin of the incision. An evaluation of the pancreatitis included a computed tomography study demonstrating an enlarged spleen twisted on its pedicle and displaced inferiorly. Esophagogastric duodenoscopy performed in evaluation of gastrointestinal bleeding showed persistent gastric varices. Multiple transfusions were required. A splenectomy was performed. The recovery was uncomplicated. CONCLUSION: In this case of a wandering spleen, pancreatitis resulted from torsion and obstruction of the tail of the pancreas within the splenic pedicle and was effectively treated with splenectomy. Thrombocytopenia occurred as a result of sequestration of platelets in the enlarged spleen. Persistent gastric varices and subsequent gastrointestinal bleeding occurred secondary to splenic vein obstruction. PMID- 12738116 TI - Wandering spleen as an asymptomatic pelvic mass. AB - BACKGROUND: Wandering spleen is caused by laxity of the ligaments surrounding the spleen and may present as an asymptomatic pelvic mass. CASE: A 20-year-old woman with a neurogenic bladder was diagnosed with a pelvic mass on a routine screening ultrasound. The mass was described as solid and multilobulated, with the lobules measuring up to 5 cm in diameter. She was completely asymptomatic. Exploratory laparotomy revealed a wandering spleen. CONCLUSION: Wandering spleen, though unusual, should be included on the differential diagnosis of patients with an asymptomatic solid pelvic mass. PMID- 12738117 TI - Prenatal diagnosis of methotrexate embryopathy. AB - BACKGROUND: Methotrexate is an antineoplastic agent used by obstetrician gynecologists for termination of early pregnancy. The drug is not always successful and is associated with a known array of malformations. CASE: We present a case of a failed pregnancy termination with methotrexate, which resulted in fetal anomalies. Ultrasound revealed absent or markedly shortened long bones, abnormal positioning of the hands, micrognathia, echogenic bowel, and a two-vessel umbilical cord. The patient elected to undergo pregnancy termination, and the ultrasound findings were confirmed at autopsy. CONCLUSION: Because of methotrexate's teratogenic potential, follow-up to confirm successful termination is necessary. Ultrasound evaluation of the fetus is indicated if pregnancy termination is unsuccessful. PMID- 12738119 TI - Trial of labor in women with transverse vaginal septa. AB - BACKGROUND: Transverse vaginal septa are rare anomalies that may be first diagnosed during pregnancy. Management options including elective cesarean delivery, incision before labor, and a trial of labor have been proposed. CASES: Two patients with transverse vaginal septa were allowed a trial of labor. The septa were incised in active labor, resulting in vaginal delivery with no related complications. CONCLUSION: Allowing a trial of labor despite a transverse vaginal septum is a reasonable management option in selected cases. PMID- 12738118 TI - Successful outcome after serial amnioreductions in triplet fetofetal transfusion syndrome. AB - BACKGROUND: Triplet fetofetal transfusion is an extremely rare complication with high perinatal mortality. Its rarity does not allow any prospective randomized study on various interventional methods to be conducted. CASE: We report one case of triplet fetofetal transfusion syndrome with survival of all three fetuses. Two were donor fetuses, and one was the recipient fetus. Serial amnioreductions were performed at 22, 24, and 26 weeks' gestation to relieve symptomatic polyhydramnios. Premature rupture of membranes occurred at 27 weeks and cesarean delivery was performed. All three babies were discharged home by 4 months of age, and all had normal neurological development when assessed at 6 months of age. CONCLUSION: The option of serial amnioreduction, with the anticipation and preparation for delivery at around 28 weeks, should be seriously considered when triplet fetofetal transfusion syndrome is encountered. PMID- 12738120 TI - Newly diagnosed adult-onset Still disease in pregnancy. AB - BACKGROUND: Adult-onset Still disease is a rare febrile disorder that can present for the first time during pregnancy. It is a diagnosis of exclusion with nonspecific laboratory results. Good maternal and fetal outcomes can be expected after initial diagnosis and treatment. CASE: A 21-year-old (gravida 2, para 1) at 20 weeks' gestation presented with fever, malaise, arthralgias, and cough. She had an extensive evaluation that led to the diagnosis of adult-onset Still disease. She was treated with prednisone and had immediate improvement. She delivered a viable infant at 34 weeks after suspected intrauterine growth restriction and continued to have recurrent symptoms postpartum. The second woman, 38 years old and gravida 3, para 1, aborta 1, presented at 22 weeks' gestation with similar symptoms and also had a similar diagnostic evaluation. She was diagnosed with adult-onset Still disease and started on prednisone, with immediate improvement, delivering a viable infant at 41 weeks' gestation. CONCLUSION: Adult-onset Still disease in pregnancy can be confused with many other diseases, but its diagnosis, after exclusion of other infectious, malignant, and rheumatic conditions, can portend good maternal and fetal outcomes. PMID- 12738121 TI - Fetal tachyarrhythmia associated with vibroacoustic stimulation. AB - BACKGROUND: Vibroacoustic stimulation is commonly used in antepartum fetal testing and has known benefits. Although the procedure has been deemed safe, questions have been raised as to potentially adverse effects, and the adult cardiology literature identifies a link between abrupt sound stimulation and severe tachyarrhythmias. CASE: A fetus with premature atrial contractions converted to supraventricular tachycardia immediately after vibroacoustic stimulation. The tachyarrhythmia spontaneously reverted to baseline after 4 minutes. The infant was treated for junctional reentry tachycardia at birth. CONCLUSION: Although the cardiac effects of vibroacoustic stimulation have previously been characterized as benign, the use of this modality may warrant restriction in the setting of known fetal arrhythmias. PMID- 12738122 TI - Autoimmune hepatitis: diagnosis after preeclampsia-induced elevated liver enzymes failed to normalize postpartum. AB - BACKGROUND: Pregnancy can unmask underlying medical conditions, as evidenced by laboratory abnormalities that do not resolve postpartum. CASE: A woman developed preeclampsia associated with elevated hepatic transaminases and serum creatinine. When her liver enzymes failed to normalize 8 weeks postpartum an evaluation led to the diagnosis of type III autoimmune hepatitis. CONCLUSION: Obstetricians should consider confirming resolution of laboratory abnormalities attributed to preeclampsia by the 6-week postpartum visit. Abnormalities that persist warrant an evaluation to exclude chronic medical conditions. PMID- 12738124 TI - Risks of rigid dilation for a radiated vaginal cuff: two related rectovaginal fistulas. AB - BACKGROUND: Dyspareunia secondary to significant radiation fibrosis of the vagina is a difficult clinical problem. The benefit of vaginal dilator therapy in such patients is unclear. CASE: We report two patients who had attempted dilation and elongation of a small, fibrotic, heavily radiated vagina and subsequently developed a rectovaginal fistula. CONCLUSION: Use of a rigid dilator in an attempt to lengthen a heavily radiated vaginal cuff may be hazardous. PMID- 12738123 TI - Erosive lichen planus of the vulva and vagina. AB - BACKGROUND: Erosive lichen planus causes erosion of the vulva and vagina and characteristic oral lesions. Dyspareunia is usual, and vaginal stenosis may occur. This report highlights the clinical features and the response to medical therapy. CASES: We report the case histories of three women who presented to the Vulvovaginal Disorders Clinic of the University of Iowa with long histories of dyspareunia and advanced vaginal scarring. In each case, the clinical diagnosis of erosive lichen planus was obvious but had not been made previously. All three women have responded well to topical treatment with tacrolimus 0.1% ointment. CONCLUSION: Erosive lichen planus should be suspected in a case of vaginal erosion or narrowing. Surgical management is inappropriate when the mucosa is eroded. Inspection of the mouth may confirm the diagnosis. PMID- 12738125 TI - Late recurrence of squamous cell cervical cancer in an episiotomy site after vaginal delivery. AB - BACKGROUND: Rarely is late recurrence from cervical cancer diagnosed at the episiotomy site. We report a case of a patient diagnosed with squamous cell cervical cancer at the time of delivery. CASE: Recurrence at the episiotomy site was noted more than 5 years after initial diagnosis of cervical cancer and was treated with surgery and radiotherapy. The patient has had no evidence of disease 54 months after treatment. CONCLUSION: After review of the literature, it appears that surgery combined with radiotherapy is recommended over single modality treatments. PMID- 12738126 TI - Eosinophilia and uterine leiomyosarcoma. AB - BACKGROUND: Tumor-associated blood eosinophilia and tumor-associated tissue eosinophilia have been described in association with a spectrum of neoplasms. A case of uterine leiomyosarcoma exhibiting both blood and tissue eosinophilia is presented. CASE: A 67-year-old postmenopausal woman was evaluated for a 6-month history of postmenopausal bleeding. An office-based endometrial biopsy yielded atypical mesenchymal tissue with marked eosinophilic infiltrate suspicious for a uterine sarcoma, and the patient was referred to the gynecologic oncology service in a tertiary care hospital. There was evidence of leukocytosis and eosinophilia in preoperative laboratory assessment. The patient underwent an exploratory laparotomy, a total abdominal hysterectomy, bilateral salpingo-oophorectomy, and pelvic lymph node dissection. Histopathology confirmed a low-grade uterine leiomyosarcoma confined to the uterus. There was extensive infiltration of the tumor with eosinophils. The systemic leukocytosis and the absolute eosinophilia responded to surgical removal of the tumor, with normalization of values in the immediate postoperative period. CONCLUSION: A differential diagnosis of malignancy should be entertained during an evaluation of systemic eosinophilia. PMID- 12738127 TI - Giant pelvic retroperitoneal leiomyoma arising from the rectal wall. AB - BACKGROUND: Pelvic retroperitoneal leiomyomas arising from the rectal wall are rare. We present a case of a giant retroperitoneal leiomyoma mimicking bilateral solid adnexal masses in a postmenopausal woman. CASE: A 54-year-old postmenopausal woman presented with a large abdominopelvic mass. At surgery, the uterus was displaced anteriorly by a large retroperitoneal mass. The rectosigmoid colon was noted to course through the retroperitoneal mass. The patient underwent complete excision of the retroperitoneal mass along with a rectosigmoid resection of the involved colon with primary reanastomosis. Histopathology showed a leiomyoma arising from the muscularis propria of the rectum wall. CONCLUSION: Retroperitoneal masses that extend into the pelvis may mimic adnexal masses and, therefore, represent a rare finding at gynecologic surgery. PMID- 12738128 TI - Fetal sex determination from maternal blood at 6 weeks of gestation when at risk for 21-hydroxylase deficiency. AB - BACKGROUND: 21-hydroxylase deficiency can lead to masculinization of female fetuses. Corticosteroid therapy may reduce these effects. When the fetus is male, this approach means that unnecessary treatment, with theoretic side effects, is given until the result of chorionic villus sampling (CVS), a procedure with known risks, is available. CASE: A woman was referred for prenatal assessment at 6 weeks' gestation because her first daughter had been born virilized from 21 hydroxylase deficiency. A real-time polymerase chain reaction assay was performed on maternal blood to detect the fetal Y chromosome-associated SRY gene. A positive signal for the SRY gene was observed. The assay was repeated a few days later, and the result was again consistent with a male fetus. CONCLUSION: Analysis of cell-free fetal deoxyribonucleic acid in maternal plasma for fetal sex determination might reduce the need for corticosteroid administration and CVS in women with fetuses at risk for 21-hydroxylase deficiency. PMID- 12738129 TI - Gestational trophoblastic tumor after medical abortion. AB - BACKGROUND: Nonmetastatic gestational trophoblastic tumor occurring after early elective medical abortion using mifepristone and misoprostol is unusual. CASE: A young woman at 6 to 7 weeks' gestation presented with brown spotting requesting medical abortion. Pretreatment ultrasound was consistent with an abnormal pregnancy. Passage of tissue ensued after mifepristone-misoprostol administration. Recovery was normal, with a postabortion ultrasound on day 16 showing a reduction in intracavitary contents. The patient declined surveillance with serial beta-human chorionic gonadotropin (beta-hCG) levels and was lost to follow-up. Sixty days after initial treatment, she presented to a hospital with a history of intermittent bleeding and underwent curettage, revealing a complete hydatidiform mole. Chemotherapy was instituted when levels of hCG plateaued. Complete hCG regression occurred after three weekly injections of methotrexate, and postmolar surveillance is uneventful to date. CONCLUSION: Gestational trophoblastic tumor may occur after early medical abortion. PMID- 12738130 TI - Gigantomastia complicating mirror syndrome in pregnancy. AB - BACKGROUND: Gigantomastia is a rare and dangerous condition in pregnancy. Although improvement after delivery is likely, postpartum aggravation is possible. To date, various pharmacological approaches have been tried, with only marginal effectiveness. Surgical intervention is often necessary. CASE: A young woman presented at 32 weeks' gestation with mirror syndrome and gigantomastia. Two years earlier she had had reduction mammoplasty by free nipple transplant. She delivered by cesarean. Rapid postpartum progression of gigantomastia led to breast necrosis and sepsis. The clinical course was complicated by acute respiratory distress syndrome and renal failure. Emergent bilateral simple mastectomy was performed, with subsequent clinical improvement. CONCLUSION: When this devastating condition occurs in pregnancy or postpartum, urgent surgical intervention may prevent potentially fatal complications. PMID- 12738131 TI - Maternal total parenteral nutrition and fetal subdural hematoma. AB - BACKGROUND: Fetal subdural hematoma is rare, and no case resulting from vitamin K deficiency secondary to maternal total parenteral nutrition has been reported. CASE: A 28-year-old woman was managed with total parenteral nutrition from 28 weeks' gestation because of continuous vomiting due to esophageal hiatal hernia. A sinusoidal pattern by cardiotocogram was observed at 31 weeks' gestation. Serial sonograms showed a fetal subdural hematoma, and cesarean delivery was performed. Although the maternal hepaplastin test result was normal and the maternal PIVKA-II concentration was only slightly elevated, the neonate was severely anemic and had severe vitamin K deficiency. CONCLUSION: Severe fetal vitamin K deficiency can develop even when the maternal deficiency is mild. When maternal total parenteral nutrition is necessary, supplemental vitamin K should be administered. PMID- 12738132 TI - Heterophile antibody blocking agent to confirm false positive serum human chorionic gonadotropin assay. AB - BACKGROUND: Persistently false positive human chorionic gonadotropin (hCG) test results in the absence of an intrauterine pregnancy may lead to a diagnosis of ectopic pregnancy or gestational trophoblastic neoplasia and unnecessary testing and therapy. CASE: A 38-year-old woman on oral contraceptives and a 53-year-old perimenopausal woman presented with persistently elevated serum beta-hCG levels from 30 to 225 mIU/mL. Both women had a history of working with mice. Urine hCG testing was negative. Serum beta-hCG levels for both patients were negative after pretreatment of their serum with a heterophile antibody blocking agent. CONCLUSION: These cases illustrate that serum heterophile antibodies can interfere with the hCG enzyme immunoassay and result in false positive values. The addition of heterophile blocking agent to the serum can exclude false positives, thereby preventing unnecessary evaluation and treatment. PMID- 12738133 TI - King syndrome in pregnancy. AB - BACKGROUND: King syndrome is characterized by the presence of a nonspecific myopathy, a susceptibility to malignant hyperthermia, and dysmorphic features similar to the phenotype seen in Noonan syndrome. CASE: A young primigravida with King syndrome presented at 19 weeks' gestation. The patient required nocturnal mechanical ventilation via tracheostomy, and there was concern that the patient would become increasingly dependent on mechanical ventilation during pregnancy. The patient underwent a forceps-assisted delivery at 35 weeks' gestation, and both the patient and her infant did well and were discharged on postpartum day 3. CONCLUSION: In the gravida with King syndrome, agents that may trigger malignant hyperthermia should be avoided. Significant myopathy may result in respiratory compromise, and a multidisciplinary approach is necessary to optimize maternal and fetal outcome. PMID- 12738135 TI - 2002 Roy M Pitkin award. PMID- 12738134 TI - Conservative treatment of a second trimester cervicoisthmic pregnancy diagnosed by magnetic resonance imaging. AB - BACKGROUND: A cervicoisthmic pregnancy, which may be carried to term, is potentially dangerous for the pregnant woman. With ultrasonographic evaluation alone the diagnosis of a cervical pregnancy in the second trimester is difficult. CASE: A nulliparous 33-year-old woman at 21 weeks' gestation was diagnosed by ultrafast T2-weighted magnetic resonance imaging (MRI) to have a cervicoisthmic pregnancy. After the infant was delivered live by cesarean with a vertical fundal incision, methotrexate was infused into the placenta via the umbilical vein. The next day she received uterine artery methotrexate infusion and embolization with platinum coils. Eight months later there was no trace of the placenta on ultrasonography or MRI. She subsequently resumed normal menstrual cycles, conceived, and delivered a healthy infant at term by cesarean after 2 years. CONCLUSION: This report describes MRI and successful preservation of fertility with such an advanced cervicoisthmic pregnancy. PMID- 12738136 TI - Aneuploidy screening: the changing scene. PMID- 12738137 TI - Medical misspellings. PMID- 12738138 TI - Commentary on "pelvic scoring for elective induction", 1964. PMID- 12738140 TI - Can amnio-polymerase chain reaction alone replace conventional cytogenetic study for women with positive biochemical screening for fetal Down syndrome? AB - To determine whether amnio-polymerase chain reaction (amnio-PCR) can replace conventional cytogenetic study for confirming the karyotype of fetuses in women with positive biochemical screening for fetal Down syndrome. To check the accuracy of this technique in our laboratory, we first compared the amnio-PCR results with those of conventional cytogenetic study in 235 patients referred from June 1999 to December 2001 for prenatal diagnosis in a referral center in Hong Kong. We then reviewed the results of 1526 amniotic fluid cultures performed for positive fetal Down syndrome screening between January 1997 and December 2001 and classified them as detectable or not detectable by amnio-PCR, using the assumption that we had replaced conventional cytogenetic study with amnio-PCR. The 235 amnio-PCR results were all informative, without a false-positive or false negative result. Of the 1526 cases with positive fetal Down syndrome screening and no ultrasound abnormalities, only two cases of sex chromosome abnormalities and two cases of marker chromosomes would have been missed if conventional cytogenetic study had been replaced by amnio-PCR.Amnio-PCR can be an alternative to conventional cytogenetic study for women with positive biochemical screening for fetal Down syndrome and no demonstrable fetal structural abnormality. PMID- 12738139 TI - Randomized clinical trial of metronidazole plus erythromycin to prevent spontaneous preterm delivery in fetal fibronectin-positive women. AB - OBJECTIVE: To estimate whether antibiotic treatment of asymptomatic women with a positive cervical or vaginal fetal fibronectin test in the second trimester would reduce the risk of spontaneous preterm delivery. METHODS: Women were screened between 21 weeks 0 days and 25 weeks 6 days of gestation with cervical or vaginal swabs for fetal fibronectin. Women with a positive test (50 ng/mL or more) were randomized to receive metronidazole (250 mg orally three times per day) and erythromycin (250 mg orally four times per day) or identical placebo pills for 10 days. The primary outcome was spontaneous delivery before 37 weeks' gestation after preterm labor or premature membrane rupture. RESULTS: A total of 16,317 women were screened for fetal fibronectin, and 6.6% had a positive test; 715 fetal fibronectin test-positive women consented to randomization. Outcome data were available for 703 women: 347 in the antibiotic group and 356 in the placebo group. The antibiotic and placebo groups were not significantly different for maternal age (P =.051), ethnicity (P =.849), marital status (P =.127), education (P =.244), and bacterial vaginosis (P =.236). No difference was observed in spontaneous preterm birth before 37 weeks' (odds ratio [OR] 1.17, 95% confidence interval [CI] 0.80, 1.70), less than 35 weeks' (OR 0.92, 95% CI 0.54, 1.56), or less than 32 weeks' (OR 1.94, 95% CI 0.83, 4.52) gestation in antibiotic- compared with placebo-treated women. Among women with a prior spontaneous preterm delivery, the rate of repeat spontaneous preterm delivery at less than 37 weeks' gestation was significantly higher in the active drug compared with the placebo group (46.7% versus 23.9%, P =.039). CONCLUSION: Treatment with metronidazole plus erythromycin of asymptomatic women with a positive cervical or vaginal fetal fibronectin test in the late second trimester does not decrease the incidence of spontaneous preterm delivery. PMID- 12738141 TI - Bacterial vaginosis, vaginal fluid neutrophil defensins, and preterm birth. AB - OBJECTIVE: To examine the association between bacterial vaginosis, vaginal fluid neutrophil defensins, and preterm birth. METHODS: Vaginal fluid specimens were obtained at 24-29 weeks' gestation from 242 cases with preterm birth and 507 noncases sampled using a case-cohort study design. We tested for bacterial vaginosis by Gram staining and Nugent scores and assayed for neutrophil defensins by enzyme-linked immunosorbent assay. Bacterial vaginosis was studied as a categoric variable (negative, intermediate, and positive), whereas defensins were studied as a continuous, categoric (based on percentiles), and dichotomous measure (presence versus absence). Three gestational age cut points were used to define preterm birth. Modified Cox proportional hazard models were used to evaluate the associations between bacterial vaginosis, defensins, and degree (less than 32, less than 34, and less than 37 weeks) and type (premature rupture of membranes, preterm labor) of preterm birth. RESULTS: Elevated vaginal fluid neutrophil defensins were not associated with birth before 37 weeks. Compared with women who did not have measurable vaginal fluid defensins, women with higher defensin levels (0-2.8 micro g/mL, 2.8-8.2 micro g/mL, and greater than 8.2 micro g/mL) had a greater risk of delivering before 32 weeks. Hazard ratios adjusted for maternal race and vaginal bleeding during pregnancy and 95% confidence intervals for these defensin levels were 1.7 (0.4, 6.9), 2.4 (0.7, 7.9), and 3.1 (1.0, 9.8), respectively. Bacterial vaginosis status did not influence the association between defensins and preterm birth. CONCLUSION: Elevated concentrations of vaginal fluid neutrophil defensins at 24-29 weeks' gestation might predict preterm birth before 32 weeks. PMID- 12738143 TI - Observer agreement with laparoscopic diagnosis of pelvic inflammatory disease using photographs. AB - OBJECTIVE: To test the intraobserver (are observers likely to agree between themselves?) and interobserver (are observers likely to agree with other observers?) reproducibility and the overall diagnostic accuracy of laparoscopic diagnosis of pelvic inflammatory disease (PID). METHODS: Three senior consultants and three residents in training in obstetrics and gynecology scored the four laparoscopic images (adnexa, cul-de-sac, and pelvic panoramic view) from each of 40 patients and repeated the process 2 days later after the order of presentation had been randomized. A standardized predesigned scoring form was used. Histopathologically proven PID was used as the gold standard. RESULTS: The overall accuracy of the laparoscopic diagnosis of PID was 78%, the sensitivity was 27%, and the specificity was 92%. The overall intraobserver reproducibility of the diagnosis of PID was only fair (kappa = 0.58), and it was clearly better among the consultants than among the residents (kappa = 0.76 and 0.39, respectively). The overall interobserver reproducibility was poor to fair (kappa = 0.43), and it was again better among the consultants than among the residents (kappa = 0.48 and 0.38, respectively). When specific diagnostic features (including tubal erythema, edema, adhesions, cul-de-sac fluid) were separately analyzed, the results were no different suggesting only poor-to-fair reproducibility. CONCLUSION: Based on photographic images, the observer reproducibility and the overall diagnostic accuracy of the laparoscopic diagnosis of PID are unsatisfactory when histopathologically proven PID is used as the gold standard. PMID- 12738142 TI - Risk factors associated with pelvic floor disorders in women undergoing surgical repair. AB - OBJECTIVE: To identify demographic, obstetric, and gynecologic risk factors associated with the development of pelvic floor disorders in women who undergo surgical correction. METHODS: We conducted a case-control study, with cases selected from all women who had surgery by our urogynecology service from July 1, 1999 to July 1, 2000 and who had a first obstetric delivery at Magee Womens Hospital (n = 80). Controls were patients seen in the general gynecology office over the same time period who had no complaints associated with pelvic floor disorders in the previous 3 years, less than stage I prolapse on pelvic examination, and first obstetric delivery at Magee Womens Hospital (n = 176). Demographic, obstetric, and gynecologic variables were compared between cases and controls. RESULTS: There were no significant differences in race, current age, gravidity, or parity. Cases were more likely than controls to have a higher body mass index (BMI) (28.6 +/- 6.3 versus 26.4 +/- 6.1 kg/m(2), P =.01), to be younger at first delivery (25.8 +/- 5.3 versus 28.4 +/- 4.9 years, P <.001), to have undergone a forceps delivery (64% versus 44%, P < orr =.001), and to have had previous gynecologic surgery (34% versus 16%, P =.003). Using logistic regression modeling, all of these factors were found to be independently associated with pelvic floor disorders. After menopause, use of hormone replacement therapy 5 or more years was protective (P =.001). CONCLUSION: In our surgical patients, younger age at first delivery, higher BMI, forceps delivery, and history of gynecologic surgery were significantly associated with subsequent development of pelvic floor disorders. PMID- 12738144 TI - Effects of race and clinical factors on short-term outcomes of abdominal myomectomy. AB - OBJECTIVE: To estimate the effects of race and preoperative uterine anatomy on complication rates after myomectomy. METHODS: A total of 239 abdominal myomectomies were performed at Duke University Medical Center from July 1992 through June 1998. Charts were abstracted using standardized forms. We assessed patient characteristics, surgical indications, preoperative hematocrit, and operative findings. Outcomes were defined as any complication, including transfusion. RESULTS: The population (n = 225) was 53% black and 47% white. The mean body mass index was 26. Fourteen percent had comorbidities. Twenty percent required transfusion. Black women were found to be more likely to have uteri with more than four leiomyomata and less likely to have only one leiomyoma (P =.001). Black women were 2.48 times more likely to have a complication (P <.006). Race was no longer a significant predictor for complications (odds ratio [OR] 1.36, 95% confidence interval [CI] 0.56, 3.15) after adjustment for uterine size (OR 1.86, 95% CI 1.3, 2.67), number of leiomyomata (OR 1.83, 95% CI 1.1, 3.14), and comorbidities (OR 2.77, 95% CI 1.1, 7.69). A similar pattern was seen for blood transfusion. CONCLUSION: Black women undergoing myomectomy are more than twice as likely to have in-hospital complication or blood transfusion than white women. This is largely attributable to differences in uterine size and leiomyoma number. Research is needed to explore why black women are more likely to have larger and more numerous leiomyomata at the time of presentation for surgery. PMID- 12738145 TI - Survival and prognostic factors in patients with ovarian cancer. AB - OBJECTIVE: To assess incidence during a 10-year study period and to identify and discuss clinical relevance for prognostic factors of survival within a cohort of Norwegian ovarian cancer patients. METHODS: Incidence and prognostic factors of survival within a population-based cohort of ovarian cancer patients from one health region in Norway were examined over the 10-year period 1987 through 1996. A total of 571 histologically verified cases of primary ovarian cancer originally registered either in the Cancer Registry of Norway or in the hospital's discharge registers were included in the study. Pearson chi(2) test was used in univariate analyses of cofactors by 5-year survival, and Kaplan-Meier survival curves were computed and tested statistically by the log rank test. A multivariable proportional hazard model (Cox) was applied to assess the prognostic significance of the different covariates. RESULTS: The incidence and crude 5-year survival remained stable over the 10-year study period. The standardized incidence rate for the time periods 1987-1991 and 1992-1996 was 11.9/100,000 and 12.5/100,000, respectively. The crude 5-year survival rate for the cohort was 39%, whereas median survival was 32 months. Cox multivariable regression analysis showed that the only independent significant prognostic factors were International Federation of Gynecology and Obstetrics stage (P <.001), size of residual tumor at the end of primary surgery (P <.001), and age at diagnosis (P <.01). Variables such as time period, histologic type and grade, treating hospital, comorbidity, or CA 125 were insignificant in predicting 5-year survival. CONCLUSION: The results underline the importance of improved surgical management of ovarian cancer, as residual tumor is the only prognostic factor achievable. PMID- 12738146 TI - Depression screening attitudes and practices among obstetrician-gynecologists. AB - OBJECTIVE: To assess obstetrician-gynecologists' attitudes and practices related to depression screening. METHOD: A total of 282 obstetrician-gynecologists completed a 36-question mail survey that assessed attitudes regarding depression screening, training to treat depression, psychosocial concern, professional influence, and ease of screening. RESULT: Depression screening (employed regardless of signs or symptoms) was reported by 44% of physicians. Positive attitudes toward depression screening, high psychosocial concern, high ease of screening, and adequate training to treat depression were significant independent predictors of depression screening practices. CONCLUSION: The majority of obstetrician-gynecologists are concerned about depression, believe depression screening is effective, and perform some degree of depression screening with their patients. However, they perceive depression screening as difficult to carry out in everyday practice, and some question whether screening improves outcomes. PMID- 12738148 TI - Absent nasal bone in the prenatal detection of fetuses with trisomy 21 in a high risk population. AB - OBJECTIVE: To estimate the usefulness of absent nasal bone by ultrasound in the prenatal detection of second-trimester fetuses with trisomy 21. METHODS: This was a matched case-control study of sonograms from January 1, 1997 to April 30, 2002. Genetic sonograms and facial profile pictures of all fetuses that were subsequently proven to have trisomy 21 were reviewed (study group). A control group was identified during the same study period by using a 4-to-1 ratio matching for gestational age at the time of the ultrasound examination. The sensitivity and specificity of absent fetal nasal bone for trisomy 21 were determined, and overlap with other ultrasound aneuploidy markers was assessed. RESULTS: Forty fetuses were identified with trisomy 21; in 29 (72.5%) a facial profile had been obtained. Of the 160 controls, 102 (64%) had facial profiles documented. Of the 29 fetuses with trisomy 21 with facial profile available, 12 had absent nasal bone (sensitivity 41%). None of the 102 control fetuses with facial profiles available had absent nasal bone (specificity 100%). The sensitivity of genetic ultrasound was increased from 83% (24 of 29) to 90% (26 of 29) by adding absent nasal bone to the other ultrasound aneuploidy markers. CONCLUSION: In the second trimester of pregnancy, absent nasal bone has a sensitivity of 41% and a specificity of 100% in detecting fetal trisomy 21. Absent fetal nasal bone may be added to the list of ultrasound aneuploidy markers evaluated during a genetic sonogram. PMID- 12738147 TI - Association between increased nuchal translucency and second trimester cardiac echogenic foci. AB - OBJECTIVE: To test the hypothesis that increased first trimester nuchal translucency is associated with isolated cardiac foci in the second trimester. METHODS: We identified all pregnancies delivered between January 1997 and June 2000. We included 7686 normal singleton fetuses who had a nuchal translucency scan and either a subsequent normal anomaly scan at 18-23 weeks' gestation (n = 7447) or isolated cardiac foci (n = 239). Fetuses were divided into two groups: normal (95th percentile or less) and increased (greater than 95th percentile) nuchal translucency. RESULTS: The prevalence of cardiac echogenic foci in fetuses with normal nuchal translucency was 218 of 7427 (2.9%; 95% confidence interval [CI] 2.6, 3.3%), whereas 21 of 259 fetuses (8.1%; 95% CI 5.1, 12.1%) with increased nuchal translucency were subsequently found to have cardiac foci. The adjusted odds ratio for cardiac echogenic foci in cases of increased nuchal translucency was 2.92 (95% CI 1.83, 4.66). CONCLUSION: An association exists between first trimester nuchal translucency and second trimester cardiac echogenic foci. Risk calculation algorithms for trisomy 21 based on nuchal translucency thickness should not use cardiac foci as an independent marker. PMID- 12738149 TI - Birth weight discordance and adverse fetal and neonatal outcomes among triplets in the United States. AB - OBJECTIVE: To examine the association between intratriplet birth weight discordance, fetal and neonatal mortality, and smallness for gestational age. METHODS: The 1995-1997 Centers for Disease Control and Prevention's Matched Multiple Birth file was used for this analysis. Birth weight discordance was calculated as the difference in birth weight between the largest and the smallest triplet's weight and expressed as percentage of the largest triplet's weight. For the middle-weight triplet, we also used the largest triplet's weight as a reference in calculating percentage birth weight discordance, which was then grouped into quintiles. RESULTS: Among 15,511 triplet live births and fetal deaths (at least 20 weeks' gestation), 35% had less than 10% birth weight discordance, 19.3% had 10-15%, 16.4% had 15-21%, 15.2% had 21-29%, and 14.1% had 29% or more. After controlling for confounders, the risk of fetal death associated with quintile V was significantly higher than that associated with quintile I for smallest (odds ratio [OR] 10.88; 95% confidence interval [CI] 4.87, 26.56), middle (OR 22.6; 95% CI 11.05, 46.3), and largest (OR 2.41; 95% CI 1.01, 5.89) triplets. Smallest and middle triplets in quintile V were more likely than quintile I triplets to be born small for gestational age (OR 26.0; 95% CI 17.1, 39.9 for smallest, and OR 13.4; 95% CI 8.01, 22.3 for middle). Birth weight discordance quintile was not associated with smallness for geatational age among largest triplets nor consistently with neonatal mortality among smallest, middle, or largest triplets. CONCLUSION: Increasing birth weight discordance was associated with increased risk of fetal death and smallness for gestational age. A birth weight discordance threshold of at least 29% should alert obstetricians for appropriate decision making. PMID- 12738150 TI - Persistent fetal occiput posterior position: obstetric outcomes. AB - OBJECTIVE: To evaluate the obstetric outcomes associated with persistent occiput posterior position of the fetal head in term laboring patients. METHODS: We performed a cohort study of 6434 consecutive, term, vertex, laboring nulliparous and multiparous patients, comparing those who delivered infants in the occiput posterior position with those who delivered in the occiput anterior position. We examined maternal demographics, labor and delivery characteristics, and maternal and neonatal outcomes. RESULTS: The prevalence of persistent occiput posterior position at delivery was 5.5% overall, 7.2% in nulliparas, and 4.0% in multiparas (P <.001). Persistent occiput posterior position was associated with shorter maternal stature and prior cesarean delivery. During labor and delivery, the occiput posterior position was associated with prolonged first and second stages of labor, oxytocin augmentation, use of epidural analgesia, chorioamnionitis, assisted vaginal delivery, third and fourth degree perineal lacerations, cesarean delivery, excessive blood loss, and postpartum infection. Newborns had lower 1 minute Apgar scores, but showed no differences in 5-minute Apgar scores, gestational age, or birth weight. CONCLUSION: Persistent occiput posterior position is associated with a higher rate of complications during labor and delivery. In our population, the chances that a laboring woman with persistent occiput posterior position will have a spontaneous vaginal delivery are only 26% for nulliparas and 57% for multiparas. PMID- 12738151 TI - Oral misoprostol for the third stage of labor: a randomized controlled trial. AB - OBJECTIVE: To compare oral misoprostol with conventional oxytocics in the management of the third stage of labor. In a controlled trial, 1574 women were randomized into four groups, as follows: Group 1 received intravenous infusion of oxytocin 10 IU plus oral misoprostol 400 micro g, followed by two doses of oral misoprostol 100 micro g 4 hours apart; group 2 received oral misoprostol 400 micro g, followed by two doses of oral misoprostol 100 micro g 4 hours apart; group 3 received intravenous infusion of oxytocin 10 IU; and group 4 received intravenous infusion of oxytocin 10 IU plus intramuscular administration of methylergonovine maleate (Methergine) 0.2 mg. The incidence of postpartum hemorrhage and decrease in hemoglobin concentration from before delivery to 24 hours postpartum were the main outcome measures. RESULTS: The primary outcome measures were similar in groups 2 and 3. The incidence of postpartum hemorrhage was 9% in group 2, compared with 3.2% in group 1 and 3.5% in group 4 (P <.01, and P =.01, respectively). There were no significant differences among the four groups regarding hemoglobin concentrations. Significantly more women needed additional oxytocin in group 2, when compared with group 4 (5.9% versus 2.2%; P =.01). The proportion of women requiring additional methylergonovine maleate was 4.8% in group 2, compared with 0.7% in group 1 and 1% in group 4 (P <.01 and P =.01, respectively). CONCLUSION: Oral misoprostol alone is as effective as oxytocin alone for the prevention of postpartum hemorrhage; it is less effective than oxytocin plus methylergonovine maleate and oral misoprostol plus oxytocin. PMID- 12738152 TI - Complications and untoward effects of the tension-free vaginal tape procedure. AB - OBJECTIVE: To report our experience with our first 350 cases of tension-free vaginal tape (TVT), specifically assessing intraoperative complications, postoperative morbidity, and untoward effects of the procedure. METHODS: Although increased numbers of reports have documented the efficacy of the TVT procedure, there are minimal data about the incidence of complications and how they are managed. We performed a retrospective review of all patients undergoing the TVT procedure over a 4-year period to report intraoperative complications (bladder perforation and excessive bleeding), postoperative complications (de novo urge incontinence, voiding dysfunction, erosion, nerve injury, urinary retention, hematoma formation), and incidence of reoperation either for voiding dysfunction or for recurrent incontinence. RESULTS: A total of 350 patients were included in the study. Fifty-five percent (194) of women underwent the TVT procedure in conjunction with other vaginal surgery, and 45% (156) underwent the TVT alone. Seventy women (20%) had previous antiincontinence surgery. Intraoperative complications included 19 bladder perforations in 17 patients (4.9%) and three cases of significant bleeding (0.9%). Postoperatively, 17 women (4.9%) had voiding dysfunction and 42 (12%) required anticholinergic therapy beyond 6 weeks. Recurrent urinary tract infections developed in 38 (10.9%), erosion or poor healing in three (0.9%), hematoma in six (1.7%), and nerve injury in three (0.9%). Twenty-eight (8%) underwent urethral dilation in the postoperative period for varied amounts of voiding dysfunction. Of these, 82% were either improved or were cured. Six women (1.7%) underwent a takedown of the TVT procedure for continued voiding dysfunction, and two of these (33%) developed recurrent stress incontinence. To date, two patients (0.5%) have undergone another procedure for recurrent or persistent stress incontinence. CONCLUSION: The TVT procedure is efficacious for the correction of stress incontinence. Our data show that it is a safe procedure with an acceptable complication rate when performed by surgeons who have experience with retropubic and transvaginal antiincontinence procedures. PMID- 12738153 TI - The relationship of tension-free vaginal tape insertion and the vascular anatomy. AB - OBJECTIVE: To describe the proximity of the major vessels in the retropubic space and anterior abdominal wall to the tension-free vaginal tape needle. METHODS: Tension-free vaginal tape needles were inserted bilaterally in ten cadavers. Dissection of the superficial epigastric, inferior epigastric, external iliac, and obturator vessels was performed. Measurements from the lateral aspect of the needle to the medial edge of the vessels were recorded. In an additional cadaver, three planes were created by placing a string from the midlabia to the shoulder, mid-biceps brachii muscle, and 6 cm lateral to the mid-biceps brachii muscle of the cadaver's extended, ipsilateral arm. An operator, blinded to the retropubic space anatomy, passed the needle in these planes bilaterally. The distances from the needle to the external iliac and obturator vessels were measured. RESULTS: All vessels measured were lateral to the tension-free vaginal tape needle. The mean distance from the tension-free vaginal tape needle to the obturator vessels was the closest: 3.2 cm (range 1.6-4.3 cm). The mean distance from the tension free vaginal tape needle to the superficial epigastric vessels was 3.9 cm (range 0.9-6.7); to the inferior epigastric vessels, 3.9 cm (range 1.9-6.6 cm); and to the external iliac vessels, 4.9 cm (range 2.9-6.2 cm). When the needle was directed 6 cm lateral to the mid-biceps brachii muscle, the external iliac vein was punctured. CONCLUSION: The major vessels in the retropubic space and anterior abdominal wall lie 0.9-6.7 cm lateral to the tension-free vaginal tape needles. If the tension-free vaginal tape needle is laterally aimed or rotated, major vascular injury can occur. PMID- 12738154 TI - Bladder tumor found by tension-free vaginal tape procedure. AB - BACKGROUND: Tension-free vaginal tape (TVT) is thought to be a most effective therapy against stress urinary incontinence. It is rare to find a bladder tumor by the TVT procedure. CASE: A 76-year-old woman with stress urinary incontinence underwent a TVT procedure. Cystoscopy performed in conjunction with the TVT procedure detected a papillary tumor. Transurethral resection of the bladder tumor was performed after the TVT procedure. CONCLUSION: Taking the possible presence of bladder tumors into consideration, TVT-related cystoscopy procedures should be performed carefully. Moreover, the case reported here suggests, at least in the short term, not only that the TVT device does not adversely affect the operability of transurethral resection of the bladder tumor, but also that the transurethral resection of the bladder tumor procedure does not lead to increased levels of urinary incontinence. PMID- 12738155 TI - Development of endometrial cancer after radiation treatment for cervical carcinoma. AB - OBJECTIVE: To detail the Memorial Sloan-Kettering Cancer Center and MD Anderson Cancer Center experience with 23 patients treated with radiation therapy for invasive cervical carcinoma who subsequently developed endometrial carcinoma. METHODS: We conducted a retrospective chart and pathology review on patients diagnosed with endometrial cancer between 1976 and 2000 who had previously received definitive radiation treatment for cervical cancer. Abstracted data included patient demographics, type of radiation therapy, histological grade, histological subtype, and stage of endometrial cancer. RESULTS: The mean age at endometrial cancer diagnosis was 64.4 years (range 53-80), and the average latency period from initial therapy to development of endometrial carcinoma was 14 years (range 6-27). Distribution by stage, grade, and histology was as follows: stage I, five (22%); stage II, one (4%); stage III, nine (39%); stage IV, seven (30%); unknown stage, one (4%); grade 1, one (4%); grade 2, three (13%); grade 3, 17 (74%); unknown grade, two (9%); carcinosarcoma, eight (35%); endometrioid, four (17%); papillary serous, six (26%); clear cell, one (4%); mucinous, one (4%); undifferentiated, one (4%); and unknown histology, two (9%). The median survival was 24 months, and the 2- and 5-year survival rates were 50% (95% confidence interval [CI] 31.4%, 78.9%) and 21% (95% CI 8.1%, 56.3%), respectively. CONCLUSION: Patients treated with definitive radiation therapy for invasive cervical cancer may still have viable endometrium at risk for neoplasia. Endometrial cancers that develop after radiation treatment have a preponderance of high-risk histological subtypes and, consequently, a poor prognosis. PMID- 12738156 TI - Lymphangioma circumscriptum of the vulva: a review of the literature. AB - OBJECTIVE: To promote proper identification and management of this rare entity by presenting a review of the literature, a case report, and illustrations of its presentation and pathology. DATA SOURCES: Sources searched included the literature contained in the National Library of Medicine's PubMed database, using the term "lymphangioma circumscriptum" combined with "vulva." Other search terms used were "capillary lymphangioma," "lymphangiectasia," and "dermal lymphangioma." Sources also included articles predating the PubMed database that were cited by other writers. METHODS OF STUDY SELECTION: Selections were restricted to the English-language medical literature published since 1960, and the search resulted in retrieval of 3272 published papers about lymphangioma. TABULATION, INTEGRATION, AND RESULTS: Congenital lymphangioma circumscriptum of the vulva has been reported in 11 patients, including one case reported and illustrated in this paper, and acquired lymphangioma circumscriptum has been reported in 20. It affects females 9-76 years old (mean 42.5 years). Clinically, it is characterized by persistent clusters of thin-walled vesicles filled with clear fluid. The diagnosis is usually made by biopsy, as these lesions often mimic such infectious diseases as molluscum contagiosum. Management options have included surgical excision of the skin and subcutaneous tissue, surface abrasion by laser or sclerosing therapy, and observation. CONCLUSION: From this comprehensive review, which includes the clinicopathologic features of both the congenital and acquired forms, illustrations from the rarest form, and a summary of treatment approaches, we conclude that lymphangioma circumscriptum poses a diagnostic challenge the risks of which are misdiagnosis and mistreatment. These risks would likely be reduced were a database of cases accessible that permitted long-term follow-up and better assessment of presenting characteristics and treatment options. PMID- 12738157 TI - A demographic analysis of the origin of papers published in Obstetrics and Gynecology. AB - OBJECTIVE: To ascertain temporal trends in the number and share of papers originating from work sites in the ten districts of The American College of Obstetricians and Gynecologists (ACOG) and to identify demographic predictors of variance among the districts. METHODS: The work sites of the first authors of papers published in Obstetrics & Gynecology were determined for selected years since 1985 and sorted by ACOG district. Three related journals (Fertility and Sterility, Gynecologic Oncology, and the American Journal of Obstetrics and Gynecology) were similarly analyzed for the year 2000. Demographic variables, including numbers of ACOG Fellows, residencies, subspecialty fellowships, and medical schools, were analyzed with multivariable regression for the most predictive variables for number of papers among ACOG districts. RESULTS: The number and share of papers published in Obstetrics & Gynecology written by authors working in ACOG districts have been declining steadily since 1985, in contrast to the number of papers arising from locations outside of ACOG districts. Analysis of demographic factors for number of papers from four specialty journals in the year 2000 revealed that the number of medical schools in the district (R(2) = 0.79) was the most predictive (P <.001). CONCLUSION: Efforts to identify and correct factors associated with a decline in the number of published papers should focus on conditions in medical schools. PMID- 12738158 TI - Spontaneous abortion among pregnancies conceived using assisted reproductive technology in the United States. AB - OBJECTIVE: To examine rates and risk factors for spontaneous abortion among pregnancies conceived using assisted reproductive technology (ART). METHODS: Subjects were 62,228 clinical pregnancies resulting from ART procedures initiated in 1996-1998 in US clinics. Spontaneous abortion was based on ART clinic report and was defined as loss of the entire pregnancy. Spontaneous abortion rates for ART pregnancies were compared with spontaneous abortion rates from the National Survey of Family Growth, a population-based survey of US women 15-44 years. RESULTS: The spontaneous abortion rate among ART pregnancies was 14.7%. This was similar to rates among pregnancies reported in the National Survey of Family Growth. Among pregnancies conceived with the patient's oocytes and freshly fertilized embryos, the spontaneous abortion risk ranged from 10.1% among women 20-29 years to 39.3% among women older than 43. Spontaneous abortion risk among pregnancies conceived with donor eggs was 13.1% with little variation by patient age. Spontaneous abortion risk was increased for pregnancies conceived with frozen and thawed embryos and decreased among multiple-gestation pregnancies. Spontaneous abortion risk was increased among women reporting previous spontaneous abortions and ART attempts, and among women who used clomiphene or zygote intrafallopian transfer. Pregnancies conceived by young women, but gestated by a surrogate, were at increased risk for spontaneous abortion in comparison with young women who gestated their own pregnancies. CONCLUSION: These findings suggest that ART does not pose a risk for spontaneous abortion. Factors related to oocyte or embryo quality are of primary importance in assessing spontaneous abortion risk. PMID- 12738159 TI - Pharmacokinetics and adverse-effect profile of rectally administered misoprostol in the third stage of labor. AB - OBJECTIVE: To characterize the pharmacokinetics and adverse-effect profile of rectally administered misoprostol. METHODS: To assess absorption of rectally administered misoprostol, 20 women were randomized to receive misoprostol 600 microg by either oral or rectal administration after delivery. Blood samples were obtained at 0, 7.5, 15, 30, 45, 60, 90, 120, and 240 minutes and analyzed for serum concentrations of misoprostol free acid by enzyme-linked immunosorbent assay. Additionally, 275 women were randomized to receive rectal 400 microg, rectal 600 microg, or oral 600 microg misoprostol after delivery. Self-assessment questionnaires were used to ascertain the adverse-effect profiles. RESULTS: Misoprostol tablets are absorbed rectally even though they are formulated for oral use. The area under the curve (integral of concentration and time graph) for rectal misoprostol was higher by 121 pg.h/mL (95% confidence interval [CI] -4.2, 246.2) than for oral misoprostol. The rectal route group had a mean maximum serum concentration 144 pg/mL lower than that for the oral route (95% CI 63, 225). This maximum was achieved on average 23 minutes later than in the oral group (95% CI 10, 35). Shivering was reported by 76% of the patients in the oral 600-microg arm, 56% of the patients in the rectal 400-microg arm, and 54% of the patients in the rectal 600-microg arm. The relative risk of shivering in the combined rectal groups is 73% that of the oral group (95% CI 61%, 86%). Severe shivering reported by patients was significantly reduced, by 72%, in rectal groups compared with the oral group (95% CI 60%, 81%). CONCLUSION: Misoprostol administered rectally is associated with lower peak levels and a reduction in adverse effects compared with the oral route. Increasing rectal doses may achieve higher efficacy without reducing the acceptability of the treatment. PMID- 12738160 TI - Placental insufficiency is characterized by platelet activation in the fetus. AB - OBJECTIVE: To investigate whether activation of circulating platelets was present in the fetal and maternal circulation in cases with vascular disease in the fetal umbilical-placental circulation as identified by umbilical artery Doppler study. METHODS: We studied 20 mother-fetus pairs with an abnormal umbilical artery Doppler study indicating umbilical-placental pathology and 9 normal pregnancy pairs. All pregnancies in these two groups had elective cesarean delivery. We also studied 15 healthy nonpregnant women. Blood was collected at delivery, and flow cytometry was used to measure platelet activation. The platelet population was specified by the antiglycoprotein IIIa (CD61) antibody and activated platelets by the anti-P selectin (CD62) antibody. Platelet activation in response to thrombin (0.03 to 0.25 U/mL) was also assessed. RESULTS: In the normal, healthy, nonpregnant women, there was no evidence of platelet activation in the fetal circulation (median, 0.63% of platelet population). Platelet activation was present in the fetal circulation in pregnancies with placental insufficiency (median, 4.57%) compared with normal pregnancies (median, 1.19%) (P =.034). The fetal platelets from pregnancies complicated by placental insufficiency also showed resistance to challenge with increasing thrombin concentration compared with normal fetal platelets (at 0.25 U/mL thrombin concentration, placental insufficiency pregnancy 69.82% and normal pregnancy 81.49%, P =.003). In the maternal circulation there were no differences in platelet activation (normal 4.89%, placental insufficiency 5.16%, P =.33) and sensitivity to thrombin challenge. CONCLUSION: In the fetal circulation, the presence of Doppler-detected umbilical-placental vascular disease was associated with significantly enhanced fetal platelet activation and resistance to thrombin challenge. These changes were not noted in the maternal circulation. This provides further evidence of a primary vascular pathology in the fetal-placental circulation independent of disease in the uteroplacental circulation when the umbilical Doppler flow velocity waveform reveals a high resistance pattern. PMID- 12738161 TI - Implementation of expedited human immunodeficiency virus testing of women delivering infants in a large New York city hospital. AB - OBJECTIVE: Since August 1999, New York has required expedited human immunodeficiency virus (HIV) testing of pregnant women in labor or their newborns, with results available within 48 hours if no intrapregnancy test result was available. We documented the frequency and circumstances of expedited HIV testing, the time required for a result to be available, and hospital factors associated with different intervals. METHODS: We conducted chart reviews for women listed in the expedited HIV testing logbook between October 1, 2000 and December 31, 2000, abstracting prenatal care history and the dates and times of hospital admission, blood specimen collection, expedited HIV testing result availability, and the infant's birth. RESULTS: Of 1115 women admitted for labor and delivery during this period, 13.6% were tested under the expedited HIV testing procedure, and none were found to be HIV positive. Twenty-seven percent of women having expedited HIV testing had documentation of testing during prenatal care that was unavailable or overlooked during admission. Expedited HIV testing results were available at 48 hours or less time for 96% of the women, although results for women admitted Friday to Sunday took longer than weekday results (mean +/- standard deviation, 30.4 +/- 11.7 hours versus 21.3 +/- 9.3 hours, P <.001). Expedited HIV testing results were available before delivery for 3.3% of women and less than 12 hours after birth for 31.7% of infants. CONCLUSION: We found excellent compliance with the 48-hour time limit for expedited HIV testing but report lapses in access to prenatal HIV testing documentation, resulting in frequent duplicative testing. Further, the potential for optimal neonatal prophylaxis within 12 hours of birth was limited, as the turnaround time for HIV results exceeded 12 hours for two thirds of the infants in our sample. PMID- 12738162 TI - Understanding the motivations, concerns, and desires of human immunodeficiency virus 1-serodiscordant couples wishing to have children through assisted reproduction. AB - OBJECTIVE: To survey the attitudes of human immunodeficiency virus (HIV) serodiscordant couples interested in assisted reproduction and better characterize their motivations for reproducing. METHODS: A prospectively designed questionnaire and open-ended interview of 50 consecutive HIV-serodiscordant couples interested in undergoing assisted reproduction to avoid transmission of virus were studied. Demographic characteristics and attitudes regarding beginning a family were obtained. By design, males were HIV seropositive (age, 38.0 +/- 5.4 years, range 26-51 years) and healthy. Women were HIV seronegative (age, 34.5 +/- 5.1 years, range 24-45 years). Most couples were married (44 of 50) and in long term relationships (duration of relationship, 8.9 +/- 4.9 years, range 1-20 years). RESULTS: Before presentation, nine of 50 couples had conceived and delivered a child (three of nine instances with knowledge of paternal HIV status). Previous timed intercourse occurred in 8% of couples (four of 50). Six individuals stated they would proceed with timed intercourse if no other alternatives existed. Forty-eight percent said they would prefer artificial insemination with donor sperm in lieu, if assisted reproduction failed or were unavailable. Forty-three percent of respondents would pursue "posthumous conception" if cryopreserved sperm or embryos were available in the event of the partner's death. Most couples discussed the possibility of single parenting (45 of 50; 90%) or the possibility for adoptive parenting (29 of 50; 58%). Couples were aware of risk, and 92% (46 of 50) understood that their child might contract HIV. CONCLUSION: Human immunodeficiency virus-serodiscordant couples are actively seeking reproductive assistance and often consider or practice unsafe measures to achieve pregnancy. Reproductive issues and concerns unique to these couples need to be addressed before treatment. PMID- 12738164 TI - D-O-G years. PMID- 12738163 TI - The evaluation and management of hirsutism. AB - Hirsutism is the presence of terminal (coarse) hairs in females in a male-like pattern, affecting between 5% and 15% of women, depending on definition. Hirsutism has a significant negative impact on psychosocial development and is usually a sign of an underlying endocrine abnormality-namely, androgen excess. The most common cause of androgen excess is the polycystic ovary syndrome (PCOS), with 21-hydroxylase-deficient nonclassic adrenal hyperplasia, the hyperandrogenic insulin-resistant acanthosis nigricans syndrome, androgen-secreting tumors, and androgenic drug intake occurring less frequently. However, although 70-80% of patients with androgen excess demonstrate hirsutism, this sign may be less prevalent among women of Asian extraction. Conversely, not all hirsute patients have evidence of detectable androgen excess, as 5-15% of these women have "idiopathic hirsutism," with normal ovulatory function and androgen levels. There is a strong familial predilection for hirsutism, primarily because the underlying endocrine disorders (eg, PCOS) and the factors regulating the development of hair growth (eg, androgen receptor activity, 5alpha-reductase activity) have a strong genetic component. The diagnostic evaluation of the potentially hirsute patient first involves confirming the presence of hirsutism and then excluding associated or etiological abnormalities and disorders (eg, ovulatory dysfunction, adrenal hyperplasia, diabetes, thyroid hormone abnormalities). Treatment should be undertaken using combination therapy, to possibly include 1) hormonal suppression (oral contraceptives, long-acting gonadotropin-releasing hormone analogues, and insulin sensitizers), 2) peripheral androgen blockade (spironolactone, flutamide, cyproterone acetate, or finasteride), and 3) mechanical/cosmetic amelioration and destruction of the unwanted hairs (electrology and, potentially, laser hair removal). The application of eflornithine hydrochloride 13.9% topical cream may also be useful to ameliorate unwanted facial hair growth. Overall, although hirsutism is a frequent and distressing abnormality often signaling an underlying endocrine disorder, a systematic approach to evaluation will uncover the etiology, and combination therapy will provide satisfactory treatment for most patients. PMID- 12738165 TI - Diagnostic performance of intracardiac echogenic foci for Down syndrome: a meta analysis. AB - OBJECTIVE: To synthesize the accumulated data on the diagnostic performance of intracardiac echogenic foci for Down syndrome, a meta-analysis was performed. DATA SOURCES: We conducted MEDLINE and EMBASE searches (1985 to August 2002) using the key words "intracardiac (echogenic) focus/foci," "golfballs," "trisomy 21," and "Down syndrome." Bibliographies of retrieved articles were also screened, and experts were contacted. Both single and multiple intracardiac echogenic foci qualified, regardless of cardiac location. Eligible studies included and described both chromosomally normal and abnormal fetuses; the fetal karyotype was unknown at the time of sonographic examination; and chromosomal status was confirmed by karyotype or postnatal clinical examination. TABULATION, INTEGRATION AND RESULTS: Sensitivity and specificity were recorded for intracardiac echogenic foci in a "combined" setting (regardless of the presence of other sonographic markers) and "isolated" setting (in the absence of other markers). Weighted estimates and summary receiver operating characteristic curves were calculated. Across 11 studies (51,831 pregnancies, 333 Down syndrome cases), random effects sensitivity and specificity were 26% (95% confidence interval 19, 34) and 95.8% (95% confidence interval 92.2, 97.8), respectively, with a positive likelihood ratio of 6.2 ("combined" setting, likelihood ratio 7.0; "isolated" setting, likelihood ratio 5.4). Summary receiver operating characteristic curves were also consistent with these values. With a 0.8% risk of amniocentesis-induced fetal loss, one fetus is lost per Down case detected when the background Down risk is 1:770. CONCLUSION: Intracardiac echogenic foci increase the risk of Down syndrome five- to seven-fold. This information should be considered in the decision making for amniocentesis in conjunction with the woman's background risk. PMID- 12738166 TI - Bleeding patterns of the hormone replacement therapies in the postmenopausal estrogen and progestin interventions trial. PMID- 12738169 TI - Screening interval and risk of invasive squamous cell cervical cancer. PMID- 12738170 TI - Local relapse in patients treated for squamous cell vulvar carcinoma: incidence and prognostic value. PMID- 12738172 TI - Prospective risk of stillbirth in multiple gestation pregnancies: a population based analysis. PMID- 12738173 TI - Effect of adding biofeedback to pelvic floor muscle training to treat urodynamic stress incontinence. PMID- 12738175 TI - Reflection. PMID- 12738178 TI - ACOG Committee Opinion. Number 283, May 2003. New U.S. Food and Drug Administration labeling on Cytotec (misoprostol) use and pregnancy. AB - On April 17, 2002, the U.S. Food and Drug Administration approved a new label for the use of Cytotec (misoprostol) during pregnancy. The new labeling does not contain claims regarding the efficacy and/or safety of Cytotec when it is used for cervical ripening for the induction of labor nor does it stipulate doses or dosing intervals. Therefore, the Committee on Obstetric Practice reminds Fellows that this agent should be used as previously recommended. PMID- 12738177 TI - ACOG Practice Bulletin. Clinical management guidelines for obstetrician gynecologist. Number 43, May 2003. Management of preterm labor. PMID- 12738179 TI - Indole-3-carbinol and 3,3'-diindolylmethane induce apoptosis in human prostate cancer cells. AB - Cruciferous vegetables contain glucobrassicin which, during metabolism, yields indole-3-carbinol (I3C). In a low pH environment I3C is converted into polymeric products, among which 3,3'-diindolylmethane (DIM) is the main one. The apoptotic effects of I3C and DIM were exhibited in human breast cancer cells. The objectives of this study were: (a) examination of the potential effects of I3C and DIM on the proliferation and induction of apoptosis in human prostate cancer cell lines with different p53 status; (b) to try to characterise the mechanism(s) involved in these effects. Our results indicate that both indole derivatives suppress the growth of these cells in a dose- and time-dependent manner, by inducing apoptosis. It appears that these indolic compounds may offer effective means against prostate cancer. Induction of apoptosis was p53-independent. Moreover, the indole derivatives employed did not affect the levels of bcl-2, bax and fasL. PMID- 12738180 TI - Hydrogen peroxide scavenging, antioxidant and anti-radical activity of some phenolic acids. AB - Some water-soluble phenolic acids were investigated as antioxidants, scavengers of hydrogen peroxide (H(2)O(2)) and scavengers of 1,1-diphenyl-2-picrylhydrazyl radical (DPPH(*)). The strongest antioxidant, scavenging of H(2)O(2) and DPPH(*) radical activity was exhibited by 3,4,5-trihydroxybenzoic (gallic) acid and 1,2,3 trihydroxybenzene (pyrogallol) with three hydroxyl groups bonded to the aromatic ring in an ortho position in relation to each other. Phenolic acids with two hydroxyl groups bonded to aromatic ring in the ortho position, such as 3,4 dihydroxycinnamic (caffeic), 3,4-dihydroxybenzoic (protocatechuic) and 2,3 dihydroxybenzoic (o-pyrocatechuic) acids, showed strong antioxidant and anti radical activity; however, it was lower than that of 3,4,5-trihydroxybenzoic acid or 1,2,3-trihydroxybenzene. 3,5-Dihydroxybenzoic (alpha-resorcylic) and 2,4 dihydroxybenzoic (beta-resorcylic) acids with two hydroxyls bonded in the meta position in relation to each other showed moderate antioxidant and low DPPH(*) and hydrogen peroxide scavenging activity. Compounds with one hydroxyl group such as 3-hydroxybenzoic, 4-hydroxyphenylacetic and 2-hydroxybenzoic (salicylic) acids, exhibited the lowest anti-radical and antioxidant activity. The results obtained show that the antioxidant and anti-radical activity of phenolic acids correlated positively with the number of hydroxyl groups bonded to the aromatic ring. The model of an ortho substitution of hydroxyl groups to the aromatic ring seems to be adequate for antioxidant and H(2)O(2) or DPPH(*) scavenging activity of phenolic acids. PMID- 12738181 TI - Inactivation of creatine kinase induced by quercetin with horseradish peroxidase and hydrogen peroxide. pro-oxidative and anti-oxidative actions of quercetin. AB - Pro-oxidative and anti-oxidative actions of quercetin were examined through inactivation of CK and inhibition of lipid peroxidation. Quercetin induced inactivation of creatine kinase (CK) during the interaction with horseradish peroxidase and hydrogen peroxide (HRP-H(2)O(2)). CK activity in heart homogenate was also reduced by quercetin with HRP-H(2)O(2). Flavonoids that have a catechol structure in the B ring, such as taxifolin, catechin and luteolin, also induced CK inactivation. These flavonoids strongly inhibited NADPH and ADP-Fe(3+) dependent microsomal lipid peroxidation. These results suggest a close relationship between pro-oxidative and anti-oxidative actions of quercetin. Electron spin resonance (ESR) signals of the quercetin radical was emitted during the interaction of quercetin with HRP-H(2)O(2) in the presence of Zn(2+) as a stabilizer. Adding CK diminished the ESR signals of quercetin radicals, suggesting CK efficiently scavenged quercetin radicals. Sulfhydryl groups and tryptophan residues in CK decreased during the interaction of quercetin with HRP H(2)O(2). The kinetic parameters of K(m) and V(max) for ADP and creatine phosphate changed rapidly, suggesting that the inactivation of CK was induced through conformational change of the enzyme. Glyceraldehyde-3-phosphate dehydrogenase had a higher sensitivity to quercetin with HRP-H(2)O(2) than CK. Quercetin radicals may mediate between pro-oxidative and anti-oxidative action. PMID- 12738182 TI - Fate of 14C-chlorpyrifos in stored soybeans and its toxicological potential to mice. AB - Grains of soybeans were treated with [ethyl-1-4C]chlorpyrifos at a dose of 15 ppm and its triplet. During the 30-week storage period, the penetration and distribution of insecticide residues were studied. The amount of surface residues on stored soybeans, internal extractables and bound residues were determined. Surface residues were found to decrease with the increase in time of storage, whereas internal residues showed a gradual increase with time to reach 40-52% of applied dose after 30 weeks. Non-extractable residues showed a relatively slow but definite increase with time. The degradation products include, in addition to the parent insecticide which constitutes 50% of the total residues, the oxygen analogue of chlorpyrifos, desethyl chlorpyrifos and 3,5,6-trichloropyridinol. Feeding mice for 90 days with a diet mixed with total internal chlorpyrifos residues in stored soybeans led to considerably inhibited plasma and red blood cells-cholinesterase activity by 78 and 46%, respectively, during the experimental period. Blood picture, liver and kidney function of treated mice were significantly affected during the feeding period. PMID- 12738183 TI - Developmental (embryo-fetal toxicity/teratogenicity) toxicity studies of synthetic crystalline lycopene in rats and rabbits. AB - Synthetic crystalline lycopene is a nutritional supplement to increase dietary intake of lycopene, an antioxidant carotenoid. Its potential oral developmental toxicity was studied in rats and rabbits. Each study included 3 control groups (water and matrix for Lycopene 10 CWD or LycoVit 10%), 3 Lycopene 10 CWD groups [500, 1500 and 3000 (rats)/2000 (rabbits) mg/kg/day] and 1 LycoVit 10% group [3000 mg/kg/day (rats)/2000 (rabbits)]. The high dosages were at maximum achievable concentrations and dosage volumes (15 and 10 ml/kg for rats and rabbits, respectively) of the highly viscous test material suspensions. Dosages were administered on gestation days (GDs) 6 through 19 (rats) or GDs 6 through 28 (rabbits). Endpoints evaluated included viability, body weight, feed consumption, necropsy observations [GD 20 (rats)/GD 29 (rabbits)], uterine contents and fetal viability, gender, body weight and morphology (skeletons double-stained). Feed consumption and weight gain were essentially unaffected in rats and rabbits, despite intubation problems in both species and reduced gastrointestinal motility and mortality in rabbits attributable to the physical properties of the gels. Neither Lycopene 10 CWD nor LycoVit 10% caused direct maternal or developmental toxicity in rats or rabbits at dosages as high as 3000 or 2000 mg/kg/day, respectively. PMID- 12738184 TI - Contrasting effects of non-starch polysaccharide and resistant starch-based diets on the disposition and excretion of the food carcinogen, 2-amino-3 methylimidazo[4,5-f]quinoline (IQ), in a rat model. AB - It has commonly been believed that increasing fibre in the diet should reduce the incidence of cancers, especially those of the colon and rectum. The earliest definitions of dietary fibre restricted the term to plant cell walls in which non starch polysaccharides are key chemical components. However, new definitions encompass a wider range of materials, including starches resistant to digestion in the colon (resistant starches). Nevertheless, most definitions require that "dietary fibres" show physiological effects considered beneficial against cancer, including enhanced laxation and faecal bulking. On theoretical grounds, such properties might be expected to dilute the concentration of any carcinogen present and move it more rapidly through the colon, thereby reducing bioavailability. We have compared the properties of two dietary fibre preparations that are primarily non-starch polysaccharides with two resistant starch preparations for effects on carcinogen disposition in a rodent model. Although both preparations enhanced laxation and faecal bulking, only the non starch polysaccharide preparation reduced carcinogen biovailability. Indeed, carcinogen biovailability was significantly enhanced by resistant starch. We suggest that there may be fundamental differences in the manner by which non starch polysaccharides or resistant starches affect carcinogen disposition, and express concern that the events seen with the resistant starches [RS] are unlikely to be beneficial with respect to protection against cancer by exogenous carcinogens. Furthermore, the data reveal that the observation of enhanced laxation and faecal bulking does not necessarily imply a reduction in carcinogen bioavailability. PMID- 12738186 TI - IARC carcinogens reported in cigarette mainstream smoke and their calculated log P values. AB - Cigarette smoke is a complex aerosol of minute liquid droplets (termed the particulate phase) suspended within a mixture of gases (CO(2), CO, NO(x), etc.) and semi-volatile compounds. The International Agency for Research on Cancer (IARC) has classified a number of the chemical constituents reported in cigarette mainstream smoke (MS) as carcinogens. Previously, we published a series of historical reviews reporting that 11 IARC Group 1 (known human), nine Group 2A (probable human) and 48 Group 2B (possible human) carcinogens have been observed in MS. Here, we expand the list of IARC classified carcinogens from 68 to 81 compounds (11 Group 1, 14 Group 2A and 56 Group 2B) reported in MS. A number of the IARC compounds reported in MS are found in the vapor phase including three Group 1, eight Group 2A and 18 Group 2B constituents. Several IARC MS compounds are found in both the vapor and particulate phases including two in Group 1, one in Group 2A and one in Group 2B. Forty-eight IARC MS carcinogens are found in the particulate phase only. Lipophilicity, as determined by the base 10 logarithm of the calculated octanol-water partition coefficient and denoted as Clog P, is reported for each of the 71 non-metallic MS IARC carcinogens. Clog P correlates with a number of biological activities including in vitro mutagenicity and carcinogenicity in rodents, and in the absence of any additional toxicological or epidemiological data, a high log P compound is more likely to be carcinogenic than a low log P compound. PMID- 12738185 TI - Safety evaluation of functional ingredients. AB - Functional ingredients are a diverse group of compounds that are intended to produce a positive effect on the health of the consumer. The term "functional" is not meant to differentiate these ingredients from other ingredients historically consumed as part of the food supply that are indeed biologically active constituents, for example, nutrients. Indeed, all foods should be considered "functional". The term functional ingredient is meant to convey the function of these new ingredients, which is to produce a positive health outcome via physiological activity in the body. Functional ingredients encompass elements of drugs, nutrients and food additives. A framework for evaluation of the safety of functional ingredients utilizes an understanding of both the current regulatory frameworks in place as well as the characteristics that define these particular ingredients. The types of studies conducted and the data generated to support safety of functional ingredients is product-specific and can include compositional analysis, structure/toxicity analysis, evaluation of historical and intended exposure, animal studies, clinical/epidemiologic studies, and evaluation of special considerations such as potential for adverse food or drug interactions. PMID- 12738187 TI - Effects of flaxseed and defatted flaxseed meal on reproduction and development in rats. AB - Flaxseed, a rich source of reportedly beneficial n-3 fatty acid and phytoestrogens, has not been thoroughly tested for reproductive effects. High levels of flaxseed (FS, 20 or 40%) or defatted flaxseed meal (FLM, 13 or 26%) added to AIN-93 diet were evaluated in a two-phase study: dosed during gestation only or during gestation and maturation in a lifetime study. At cesarean section on gestation day 20, neither FS nor FLM affected fertility, body weight gain, litter size, or fetal development. FLM, but not FS, decreased gestation length. The offspring of dams allowed to litter were observed to postnatal day (PND) 21 or 90. Neither FS nor FLM affected PND 21 survival indices of F1 pups. FS (20 and 40%), but not FLM, increased the anogenital index (AGI) of F1 females at PND 21. The AGI of F1 males was not affected by either FS or FLM. FLM (13 and 26%), but not FS, delayed puberty in F1 males. Age and weight at the onset of puberty in females were not affected by FS or FLM. FS and FLM caused dose-related increases in the number of F1 females with irregular estrous cycles. During PND 21-90, F1 females fed 20% FS, 13% FLM, or 26% FLM gained more weight than the controls. FS and FLM decreased thymus/body weight and thymus/brain weight ratios in weanling F1 males and females. FS and FLM decreased liver/body weight and liver/brain weight ratios in weanling F1 females, and 26% FLM decreased the same two ratios in F1 males. In conclusion, FS did not affect fetal development but did affect indices of postnatal development such as the estrous cycle. PMID- 12738188 TI - Developmental effects of serum from flaxseed-fed rats on cultured rat embryos. AB - Gestation day 9.5 rat embryos were cultured for 45 h in serum obtained from pregnant rats that had been fed throughout gestation with either a control diet (based on the AIN-93 formulation), a diet supplemented with flaxseed (20% or 40%, w/w), or a diet supplemented with de-fatted flaxseed ("flaxseed meal", 13 or 26%, w/w). The embryos were fixed in neutral formalin at the end of culture. Overall growth and development was assessed, and the presence of abnormalities was noted. A significant inhibition of growth (as determined by crown-rump length) relative to control was observed in embryos cultured in serum from rats fed the 20% flaxseed diet. The incidence of spontaneous heart inversions was increased significantly in the embryos cultured in serum from the 20% flaxseed and 26% flaxseed meal fed rats. The incidence of flexion defects was increased significantly in embryos cultured in serum from 20% flaxseed-fed rats. The lack of an apparent dose response in any of the statistically significant effects suggests that the observed anomalies were chance occurrences unrelated to the treatment group from which serum was obtained. It is therefore concluded that diets high in flaxseed or flaxseed meal do not result in serum factors that are directly embryotoxic to organogenesis-staged rat embryos. This finding is consistent with the findings of a parallel in vivo rat teratology study where no significant embryotoxicity attributable to flaxseed exposure was observed. PMID- 12738189 TI - Flaxseed increased alpha-linolenic and eicosapentaenoic acid and decreased arachidonic acid in serum and tissues of rat dams and offspring. AB - The effects of dietary flaxseed (FS), and defatted flaxseed meal (FLM) on serum and tissue fatty acid profiles were investigated. Pregnant Sprague-Dawley rats were fed AIN-93 based diets balanced in calories, fat, nitrogen, and fiber. Diets contained 0, 20%, 40% FS or 13% or 26% FLM by weight. The control, FS and FLM diets differed in linoleic acid to alpha-linolenic acid (ALA) fatty acid ratio. These diets were fed continuously during gestation, suckling period and 8 weeks post-weaning (F(1)). FS fatty acids were bioavailable and metabolized by pregnant and F(1) rats. ALA and eicosapentaenoic acid increased; linoleic and arachidonic acid decreased; and docosahexaeonic acid was unchanged in serum, 'gastric milk' and liver of FS and FLM-fed pregnant and F(1) rats. FS more than FLM, changed fatty acids profiles, but FLM and 40% FS significantly reduced serum cholesterol. Dietary 40% FS may have increased oxidative stress as evidenced by a reduction in liver vitamin E. PMID- 12738190 TI - Decreasing allergic contact dermatitis frequency through dermatotoxicologic and epidemiologic based intervention? AB - Allergic contact dermatitis (ACD) affects millions of people worldwide. In an attempt to decrease the incidence of contact dermatitis, several countries have implemented legislations to reduce the population's exposure to known sensitizers. To determine whether these legislations have been effective, we reviewed several studies examining trends in patch test reactivity before and after implementation of these legislations. Common sensitizers studied include nickel, gluceryl monothioglycolate fragrances, chromate, and thiuram. Overall, the evidence suggests a decreasing trend of ACD with appropriate formulation changes, however some of the data may be confounded by biases, and additionally, exposure to the sensitizer may still exist via non-compliance to regulations by small manufacturers. In the future population based studies should be conducted to more accurately assess trends in ACD. PMID- 12738191 TI - Induction of the procarcinogen-activating CYP1A2 by a herbal dietary supplement in rats and humans. AB - Herbal dietary supplements to promote health may be double-edge swords. A herbal dietary supplement, FastOne, which contains extracts of kola nut, grape, green tea and Ginkgo biloba, and is used as an agent for weight management, was administered to rats to test whether it induced CYP1A2, a procarcinogen activating enzyme. Western blot analysis indicated that treatments with 0.15, 0.3, 0.5, 1 and 2 g/kg of the supplement for 3 days increased CYP1A2 expression in rat liver microsomes in a dose-dependent manner. The 0.3, 0.5, 1 and 2 g/kg treatments increased rat liver microsomal CYP1A2 activity measures as the conversion of caffeine to paraxanthine to 166, 212, 331 and 473% of normal, respectively. In humans, the intake of 2 and 4 capsules of the supplement for 3 days increased CYP1A2 activity to 194 and 203%, respectively, as assessed by the change in the urinary ratio of 1,7-dimethylxanthine plus paraxanthine to unmetabolized caffeine. Intake of the herbal supplement increased CYP1A2 activity to levels higher than that observed from smoking (179%). This study suggests that the long-term intake of the dietary supplement inducing CYP1A2 may increase the incidence of colorectal cancers caused by procarcinogens activated by CYP1A2 in rapid N-acetyltransferase-2 acetylators and of lung adenocarcinoma in slow acetylators. PMID- 12738192 TI - Safety evaluation of an extract from Salacia oblonga. AB - Plant extracts from the Salacia genus have been found to have intestinal alpha glucosidase inhibitor activity, which may have application to the development of medical foods for people with diabetes. We evaluated the safety of a hot water extract of S. oblonga (salacinol extract) supplemented to or processed into a medical food. Thirty male Sprague-Dawley rats were assigned among one of three treatments: (1) EN-0178 (control, liquid diet), (2) EN-0178+salacinol (as 1 plus 500 mg of salacinol extract per 253 g diet, which was added to product immediately prior to feeding), (3) EN-0195 (as 1 plus 500 mg of salacinol extract per 253 g diet, which was added during product manufacture). After 14 days of free access to dietary treatments, rats were sacrificed, blood collected and organs weighed. Rats consuming salacinol extract had reduced (P <0.05) weight gain and feed intake. The relative (% of body weight) testicular weight was higher (P<0.05) for rats consuming salacinol extract, whereas, the relative liver and spleen weight was lower (P<0.05) for rats consuming salacinol extract. Of the serum chemistries analyzed, blood urea nitrogen and alkaline phosphatase was lower (P<0.05) for rats consuming salacinol extract. No differences in blood hematology were found. We conclude that salacinol extract, in a medical food consumed for 2 weeks in amounts estimated at 10-fold greater than proposed for human intake, did not result in clinical chemistry or histopathologic indications of toxic effects in male Sprague-Dawley rats. PMID- 12738193 TI - Absorption and metabolism of glycosidic sweeteners of stevia mixture and their aglycone, steviol, in rats and humans. AB - Stevia mixture, sweeteners extracted from the leaves of Stevia rebaudiana Bertoni, consists mainly of the glycosides of the diterpene derivative steviol. The aims of this study were to investigate the absorption (in rats) and the hepatic metabolism (in rats and humans) of both stevia mixture and steviol. Absorption was investigated both in vivo and ex vivo. In ex vivo experiments using the rat everted sac method, no absorption of stevia mixture was observed, but significant absorption of steviol was noted (equivalent to approximately 70% of the absorption reference- salicylic acid- value). In the in vivo experiment, rats received a single oral administration of either steviol or stevia mixture; a peak steviol concentration in plasma was observed 15 min after its oral administration, demonstrating rapid absorption. However, after oral administration of stevia mixture, the steviol concentration in plasma increased steadily over 8 h, suggesting that stevia mixture components are first degraded and then absorbed as steviol in the rat intestine. Steviol metabolism in humans and rats was examined by incubating steviol with liver microsomes from the two species. Oxidative (monohydroxy and dihydroxy) metabolites of steviol were observed by LC-ESI/MS after incubation with both human and rat liver microsomes. The intrinsic clearance of steviol in human liver microsomes was 4-times lower than that found in rat liver microsomes. In conclusion, this study suggests that there are no major species differences in steviol hepatic metabolism between rats and humans. Absorption from the human intestine can be predicted to occur in an analogous manner to that from the rat intestine. PMID- 12738194 TI - In vitro and in vivo percutaneous absorption of catechol. AB - The Cosmetic Ingredient Review Expert Panel found insufficient data to conclude that catechol could be used safely in permanent hair dye products. Information was lacking on the extent of oxidation and skin absorption of remaining catechol. In vitro percutaneous absorption studies were conducted in human and rat skin using a consumer permanent hair dye spiked with 0.6% catechol. A 30-min application demonstrated 0.4% of the applied dose was absorbed through human skin and 0.2% through rat skin. The minimal absorption observed was due to the short exposure time and to partial oxidation of catechol by the dye developer. The fate of catechol remaining in rat skin after exposure in vitro and in vivo was investigated with additional absorption studies using catechol in ethanol. At 72 h, 24-h application of 4% catechol resulted in skin absorption of 81% of the applied dose in vitro and 53% in vivo. Skin levels measured at 24 h remained unchanged after 72 h. Therefore the skin reservoir did not contribute to the estimated systemic absorption. A deconvolution technique employed to predict skin absorption using plasma levels from intravenous and dermal administration overestimated in vivo skin absorption due to volatility of catechol in an ethanolic vehicle. PMID- 12738195 TI - Safety evaluation of sources of docosahexaenoic acid and arachidonic acid for use in infant formulas in newborn piglets. AB - Human milk provides small quantities of preformed docosahexaenoic acid (DHA) and arachidonic acid (ARA), usually less than 1% of total fatty acids. Vegetable oil blends commonly used in infant formulas have, until recently, provided the essential fatty acid precursors for these long-chain polyunsaturated fatty acids (LCPUFA), but no preformed DHA and ARA. This study evaluated the safety of ingredient sources of DHA and ARA for use in infant formulas in a neonatal piglet model. Newborn piglets were allowed to suckle for 3 days and then divided into 4 feeding groups of 6 males and 6 females. Piglets were bottle-fed at frequent feeding intervals until 19 days of age. The composition of the piglet formulas was modeled after standard milk-based formulas for human infants while meeting nutritional requirements for piglets. Formulas were a control formula (no added DHA or ARA), a DHA formula providing 55 mg DHA/100 Cal, an ARA formula providing 96 mg/100 Cal ARA, and a DHA+ARA formula providing 34 mg DHA and 62 mg ARA/100 Cal. All formulas were equal in fat content and provided approximately 1000 Cal/l. The ARA-rich oil was from a fermentation product of Mortierella alpina (40 wt.% fatty acids as ARA) and DHA was from high DHA tuna oil (25 wt.% fatty acids as DHA). There were no test article related effects of DHA and/or ARA indicative of an adverse health consequence to the animals seen in the clinical signs, body weights, food consumption, clinical chemistry, hematology, organ weights or gross or histopathology. The findings in this neonatal animal study support the safety of these ingredient oil sources of DHA and ARA for use in infant formulas. PMID- 12738196 TI - Impact of high flaxseed diet on mitogen-induced proliferation, IL-2 production, cell subsets and fatty acid composition of spleen cells from pregnant and F1 generation Sprague-Dawley rats. AB - Flaxseed (FS) being rich in alpha-linolenic acid may alter the immune parameters. Therefore, we assessed the impact of FS and defatted flaxseed meal (FLM) on fatty acid composition, cell subsets, proliferation and IL-2 production by splenic lymphocytes. Pregnant female Sprague-Dawley rats were fed diets containing 0% FS and FLM, 20 or 40% FS, 13 or 26% FLM during gestation or gestation, lactation and 8 week post-weaning period. FS and FLM resulted in up to 8.3 fold and 4.6 fold increase in splenic ALA among pregnant rats, 4.5 fold and 1.2 fold increase in splenic ALA among F(1) generation rats. Splenic linoleic acid (LA) and arachidonic acid (AA) were 18 and 40% lower in 40% FS fed pregnant rats, and AA was 15% lower in all the other groups. Among F(1) rats, splenic LA and AA were 16 and 48% lower in 40% FS group, and AA was 18% lower in 20% FS and 26% FLM groups. Concanavalin A and phytohemagglutinin mediated proliferation of spleen cells were 60 and 52% lower in 40% FS fed pregnant and F(1) generation rats, respectively. No significant changes were observed in the cell subsets or IL-2 production by splenic cells from different groups. PMID- 12738197 TI - Mineral content in legumes and nuts: contribution to the Spanish dietary intake. AB - We evaluated the levels of essential elements as Cu, Cr, Fe and Zn, and toxic elements as Al, Ni, Pb and Cd in a total of 40 samples of different legumes and 56 samples of different nuts, that are widely consumed in Spain. These elements were determined in the samples mineralized with HNO(3) and V(2)O(5), using electrothermal atomic absorption spectrometry (ETAAS) as the analytical technique. Reliability of the procedure was checked by analysis of a certified reference material. No matrix effects were observed and aqueous standard solutions were used for calibration. In legumes, the levels ranged from 1.5-5.0 microg Cu/g, 0.05-0.60 microg Cr/g, 18.8-82.4 microg Fe/g, 32.6-70.2 microg Zn/g, 2.7-45.8 microg Al/g, 0.02-0.35 microg Ni/g, 0.32-0.70 microg Pb/g and not detectable-0.018 microg Cd/g. In nuts, the levels ranged from 4.0-25.6 microg Cu/g, 0.25-1.05 microg Cr/g, 7.3-75.6 microg Fe/g, 25.6-69.0 microg Zn/g, 1.2 20.1 microg Al/g, 0.10-0.64 microg Ni/g, 0.14-0.39 microg Pb/g, and not detectable-0.018 microg Cd/g. A direct statistical correlation between Cu-Cr, Zn Al and Cr-Ni (P<0.05), and Al-Pb (P<0.001), has been found. The growing popularity of these products in recent years on the basis of its nutritional properties and beneficial effects, requires additional data and a periodical control. In addition, the present findings are of potential use to food composition tables. PMID- 12738198 TI - Survey of arsenic and other heavy metals in food composites and drinking water and estimation of dietary intake by the villagers from an arsenic-affected area of West Bengal, India. AB - An investigation of arsenic, copper, nickel, manganese, zinc and selenium concentration in foodstuffs and drinking water, collected from 34 families and estimation of the average daily dietary intake were carried out in the arsenic affected areas of the Jalangi and Domkal blocks, Murshidabad district, West Bengal where arsenic-contaminated groundwater (mean: 0.11 mg/l, n=34) is the main source for drinking. The shallow large diameter tubewells, installed for agricultural irrigation contain an appreciable amount of arsenic (mean: 0.094 mg/l, n=10). So some arsenic can be expected in the food chain and food cultivated in this area. Most of the individual food composites contain a considerable amount of arsenic. The mean arsenic levels in food categories are vegetables (20.9 and 21.2 microg/kg), cereals and bakery goods (130 and 179 microg/kg) and spices (133 and 202 microg/kg) for the Jalangi and Domkal blocks, respectively. For all other heavy metals, the observed mean concentration values are mostly in good agreement with the reported values around the world (except higher zinc in cereals). The provisional tolerable daily intake value of inorganic arsenic microg/kg body wt./day) is: for adult males (11.8 and 9.4); adult females (13.9 and 11); and children (15.3 and 12) in the Jalangi and Domkal blocks, respectively (according to FAO/WHO report, the value is 2.1 microg/kg body wt./day). According to WHO, intake of 1.0 mg of inorganic arsenic per day may give rise to skin lesions within a few years. The average daily dietary intake of copper, nickel and manganese is high, whereas for zinc, the value is low (for adult males: 8.34 and 10.2 mg/day; adult females: 8.26 and 10.3 mg/day; and children: 4.59 and 5.66 mg/day) in the Jalangi and Domkal blocks, respectively, compared to the recommended dietary allowance of zinc for adult males, adult females and children (15, 12 and 10 mg/day, respectively). The average daily dietary intake of selenium microg/kg body wt./day) is on the lower side for the children (1.07 and 1.22), comparable for the adult males (0.81 and 0.95) and slightly on the higher side for the adult females (1.08 and 1.26), compared to the recommended value (1.7 and 0.9 microg/kg body wt./day for infants and adults, respectively). PMID- 12738199 TI - Accumulation of urinary 2-thiothiazolidine-4-carboxylic acid (TTCA) among workers occupationally exposed to carbon disulfide for 1 week. AB - OBJECTIVES: To investigate if carbon disulfide (CS(2)) accumulates after a 1-week exposure period, and how the work-shift duration and exposure magnitude affects this accumulation for the workers in viscose rayon industry. METHODS: Six 8-h and seven 12-h workers in the spinning department historically known to be exposed to high air CS(2) were recruited as the exposed groups. Seven workers from other non CS(2)-exposed departments were recruited as non-exposure controls. Exposure monitoring covered a full work shift with personal breathing zone monitoring. Urine was collected pre- and post-shift every day throughout the 5 consecutive days. 2-Thiothiazolidine-4-carboxylic acid levels in the urine (U-TTCA) were determined. RESULTS: No detectable values were found for airborne (<0.6 ppm) and urinary (<35 ng/ml) monitoring for the control groups. The exposure levels for a 12-h shift (11.3+/-1.47) (AM+/-S.D.) were significantly greater than for an 8-h shift (6.3+/-0.64). The linear accumulation trend for daily U-TTCA across the workdays was only significant for the 12-h shift at pre-shift. Statistical significance was found in the regression of the ratios for pre-shift U-TTCA to airborne CS(2) levels on the preceding day to the day of the exposure at pre shift for a 12-h shift (r=0.98, P=0.02). CONCLUSIONS: The U-TTCA accumulation for occupational exposure to CS(2) was exposure-magnitude-dependent. The linear equations derived in this study indicated that the U-TTCA increment at pre-shift for each additional daily 12-h exposure, after an adjustment for the CS(2) exposure level, was 0.02 mg/g creatinine/ppm of CS(2). The long-term exposure response under such repeated and intermittent conditions should be noteworthy. PMID- 12738200 TI - Effects of low-dose cadmium exposure on biological examinations. AB - We conducted an epidemiological study to investigate the effects of low-dose cadmium (Cd) exposure on human health in a specific area of a town in Japan where low Cd concentration was detected in rice. We compared clinical findings, urinary and whole blood Cd concentrations, and indicators of renal dysfunction between the polluted area and the control area. The study employed 44 men and 54 women from the polluted area and 21 men and 29 women from the control area. In urine analysis, as indicators of Cd exposure and possible related renal dysfunction, Cd, beta(2)-microglobulin (beta(2)-MG), alpha(1)-microglobulin (alpha(1)-MG), N acetyl-beta-D-glucosaminidase (NAG), total protein, inorganic phosphorus, lysozyme and creatinine were quantitatively measured. In blood analysis, serum IP and creatinine and whole blood Cd were measured. No case of renal dysfunction due to Cd exposure was confirmed. However, both the urinary and whole blood Cd of the polluted area were significantly higher than those of the control area for both sexes. Urinary beta(2)-MG did not differ between the two areas. For women, urinary alpha(1)-MG was significantly higher in the polluted area than in the control area. In correlation analysis, beta(2)-MG, alpha(1)-MG and NAG, were positively correlated with both of urinary and whole blood Cd for men and women in the polluted area except for between urinary beta(2)-MG and urinary Cd for men. In the control area, the sole positive correlation observed was between urinary beta(2)-MG and whole blood Cd for men. We then examined the determinants of variations of parameters in urinary and blood tests. Potential determinants were age, sex, body mass index, an indicator of smoking habits (cigarette index) and the index of estimated Cd intake from rice (Cd-rice-index). Cd-rice-index was expressed as the product of Cd concentrations in homegrown rice multiplied by daily frequency multiplied by duration (years) of residence in the polluted area. In multiple regression analysis, whole blood Cd was independently associated with Cd-rice-index, age and gender. Variations in whole blood Cd accounted for a substantial portion of the variations in urinary Cd, although they were less influential in older individuals. Whole blood Cd was the sole independent variable related to variations in urinary beta(2)-MG. Cd-rice-index accounted for a portion of the variance in urinary NAG, while age was a more powerful determinant. It was thus revealed that the consumption of homegrown rice polluted with Cd in low concentration resulted in an elevation of whole blood Cd level and consequent increase in urinary Cd level. However, it was not clearly elucidated that the excretion of urinary low-molecular microglobulins could increase significantly in response to slight elevation of Cd body load. PMID- 12738201 TI - Blood mercury following DMPS administration to subjects with and without dental amalgam. AB - The use of DMPS as a diagnostic tool in patients with symptoms allegedly caused by mercury from dental amalgam fillings is disputed. We have previously shown that the mercury concentrations in urine cannot be used in such a way. In the present study, we wished to evaluate the effect on blood mercury levels (B-Hg) following intravenously injected DMPS in four groups of subjects: 19 controls without amalgam experience; 21 healthy controls with amalgam fillings; 20 patients with self-reported symptoms from existing dental amalgams; and 20 patients who had removed amalgam fillings. A single dose of DMPS (2 mg/kg) was injected. Blood samples were collected prior to the injection and after 15, 30, 120 min, and after 24 h, and mercury was analyzed by cold vapor atomic absorption spectrophotometry. All groups showed an initial drop of 24 to 30% in the blood levels, approaching baseline values (2.5-5.5 microg/l) after 2 h. The subjects with no amalgam experience had the lowest mercury values. There was no significant difference between the three groups with such experience. There were no significant differences between the two groups with amalgam fillings present. Patients with symptoms allegedly caused by amalgam were not different from the control groups. There were indications that part of the urinary mercury excreted during the first 30 min originated from blood. PMID- 12738202 TI - Environmental benzene exposure assessment for parent-child pairs in Rouen, France. AB - There is a lack of data on environmental benzene exposure in children. In this study, we compared personal benzene exposure and inhalation uptake in a group of children to those of their parents. We also compared levels of urinary benzene metabolites, trans,trans-muconic acid (MA) and hydroquinone (HQ), for those two groups, and assessed the correlation between personal benzene exposure and urinary MA and HQ concentrations. The study was performed on 21, 2-3-year-old children and their parents recruited on a voluntary basis among non-smokers from the three largest day-care centers of the town of Rouen in France. Average benzene concentrations were measured over 5 consecutive days with diffusive samplers. The following simultaneous measurements were carried out: personal exposure of the parents, concentrations inside and outside the day care centers, and inside the volunteer's bedrooms. Morning and evening urine samples were collected during the same period. Benzene personal exposure levels were 14.4+/ 7.7 microg/m(3) and 11.09+/-6.15 microg/m(3) in parents and children, respectively. Benzene inhalation uptake estimates were 2.51+/-1.23 microg/kg/day in the group of parents and 5.68+/-3.17 microg/kg/day in the group of children. Detectable levels of MA and HQ were found in 85% and 100% of the samples, respectively. Intra-individual variation of urinary MA and HQ concentrations expressed as a coefficient of variation (CV) ranged from 63 to 232% and from 13 to 144%, respectively. Mean values of MA and HQ (in mg/g creatinine) were 1.6- and 1.8-fold higher in the group of children than in the group of parents (P=0.008 and P<0.0001, respectively). Significant correlations between metabolites levels and benzene were not found. PMID- 12738203 TI - Arsenic and heavy metal contamination of vegetables grown in Samta village, Bangladesh. AB - Drinking of arsenic (As) contaminated well water has become a serious threat to the health of many millions in Bangladesh. However, the implications of contamination of agricultural soils from long-term irrigation with As contaminated groundwater for phyto-accumulation in food crops, and thence dietary exposure to As, and other metals, has not been assessed previously in Bangladesh. Various vegetables were sampled in Samta village in the Jessore district of Bangladesh, and screened for As, Cd, Pb, Cu and Zn by inductively coupled plasma emission spectrometry (ICP-AES) and inductively coupled plasma mass spectrometry (ICP-MS). These local food products are the basis of human nutrition in this region and of great relevance to human health. The results revealed that the individual vegetables containing the highest mean As concentrations microg x g( 1)) are snake gourd (0.489), ghotkol (0.446), taro (0.440), green papaya (0.389), elephant foot (0.338) and Bottle ground leaf (0.306), respectively. The As concentration in fleshy vegetable material is low. In general, the data show the potential for some vegetables to accumulate heavy metals with concentrations of Pb greater than Cd. Some vegetables such as bottle ground leaf, ghotkol, taro, eddoe and elephant foot had much higher concentrations of Pb. Other leafy and root vegetables, contained higher concentrations of Zn and Cu. Bioconcentration factors (BCF) values, based on dry weight, were below 1 for all metals. In most cases, BCF values decreased with increasing metal concentrations in the soil. From the heavily As-contaminated village in Samta, BCF values for As in ladies finger, potato, ash gourd, brinjal, green papaya, ghotkol and snake gourd were 0.001, 0.006, 0.006, 0.014, 0.030, 0.034 and 0.038, respectively. Considering the average daily intake of fresh vegetables per person per day is only 130 g, all the vegetables grown at Samta had Pb concentrations that would be a health hazard for human consumption. Although the total As in the vegetables was less than the recommended maximum intake of As, it still provides a significant additional source of As in the diet. PMID- 12738204 TI - Iodine-129 and caesium-137 in Chernobyl contaminated soil and their chemical fractionation. AB - Soil samples from areas in Belarus, Russia and Sweden contaminated by the Chernobyl accident were analysed for (129)I by radiochemical neutron activation analysis, as well as for 137Cs by gamma-spectrometry. The atomic ratio of (129)I/137Cs in the upper layer of the examined soil cores ranged from 0.10 to 0.30, with an average of 0.18, and no correlation between (129)I/137Cs ratio and the distance from Chernobyl reactor to sampling location was observed. It seems feasible to use the (129)I/137Cs ratio to reconstruct the deposition pattern of 131I in these areas. The association of (129)I and 137Cs in the Chernobyl soil and Irish Sea sediment was investigated by a sequential extraction method. Similar speciation of (129)I in the Chernobyl soil and Irish Sea sediment was found. Approximately 70% of (129)I is bound to oxides and organic matter, and 10 20% is in the readily available phase, while most of the 137Cs (73%) in Chernobyl soil remains in the extraction residue. PMID- 12738205 TI - Micromorphology, composition and origin of the orange patina on the marble surfaces of Propylaea (Acropolis, Athens). AB - This paper is an attempt to contribute to the discussion started in the 1980s and continuing until today, on the origin of orange-brown patinas observed on marble surfaces. SEM and XRD were used in the study of the orange-brown patina growing at the east stoa of Propylaea, at the Acropolis of Athens. The patina consists of an extremely compact, homogenous, 500-microm-thick layer, mainly of hydroxyapatite. The outer part (50 microm) of the layer presents an orange colour attributed to iron in clay particles, which were deposited on the surface of the patina and have gradually been diffused in the bulk. The interface between the phosphatic layer and the marble is very uneven and full of cavities colonised by microorganisms. The phosphatic layer penetrates into the grain boundaries and fills the empty space between the grains. The presence of biological attack at the interface causes progressive erosion and detachment of both the patina and the marble. The minimum presence of oxalates, the absence of microorganisms or remnants of them in the phosphatic layer, the homogeneity and compactness of the patina, give evidence against the theory of biogenic origin. It is suggested that the layer originates from an artificial coating deliberately applied onto the marble surface for protective or aesthetic reasons, when the marble had already been weathered. Possible manmade pastes are suggested. PMID- 12738206 TI - Palladium and platinum concentration in soils from the Napoli metropolitan area, Italy: possible effects of catalytic exhausts. AB - Soils from the Napoli metropolitan area (Italy) were evaluated for Pt and Pd concentrations. One hundred and ninety-five (195) samples were collected from residual soils and non-residual soils from flower-beds in a 120 km(2) area on a 0.5 km grid in the downtown-urbanized area and on 1 km grid in suburban zones. The soils <100 mesh size fraction (150 microm) was analyzed for Pt, Pd and 37 other elements by ICP-ES and ICP-MS after aqua regia digestion. Pt and Pd contents range between <2 and 52 microg/kg and between <10 and 110 microg/kg, respectively. A large number of samples from the metropolitan area were characterized by anomalous values for Pt (>6 microg/kg) and Pd (>17 microg/kg). Non-residual soils from flower-beds are located mainly in the urbanized downtown areas subject to heavy traffic and have higher Pt and Pd contents than residual soils from suburban areas. Geochemical maps show a strong correlation between roads with major traffic flow and high Pt and Pd concentrations. In addition, data from most of the downtown flower-beds fall within the three-way catalytic converters (TWC) field identified by [Ely JC, et al. Environ Sci Technol; 35:3816 3822]. This suggests that emissions of abraded fragments from vehicle exhausts may be the source of the high values and geographic distributions of Pd and especially Pt in urban soils of Napoli. Catalytic converters (Pt/Pd/Rh) have been mandatory for gasoline-powered vehicles in Europe since 1993. Italy permitted the use of non-catalytic motor until January, 2002. This is responsible for the high values for both Pt and Pd in the non-residual soils of the urban areas of Napoli. PMID- 12738207 TI - Integrated ambient ozone evaluation by passive samplers and clover biomonitoring mini-stations. AB - An ozone monitoring network was set up using passive samplers and biological mini stations of two clones of white clover (Trifolium repens L. cv. Regal), NC-R (O(3)-resistant) and NC-S (O(3)-sensitive). This paper reports on a pilot study performed in the period June-October 1999 in the Rome municipal area by using five biomonitoring mini-stations and ozone passive samplers with a new nitrite based design. This combined methodology can be used to obtain information on the biological implications of the injury due to tropospheric ozone. The two techniques can integrate data for the short-medium period and can be placed in different urban and rural sites, proving to be a very useful tool for ozone concentration mapping. PMID- 12738208 TI - Commuter exposure to aromatic VOCs in public transportation modes in Hong Kong. AB - This study investigated commuter exposure to aromatic volatile organic compounds (VOCs) in different commuting microenvironments. In Hong Kong, more than 90% of the local citizens rely on public transport facilities in their daily commutes. During five winter months in late 2001 and early 2002, in-vehicle monitoring was performed in nine popular public transportation modes: tram, public light bus, air-conditioned bus, non-air-conditioned bus, taxi, ferry and three railway systems (Mass Transit Railway-MTR, Kowloon-Canton Railway-KCR and Light Rail Transit-LRT). These transports were grouped into three categories: railway transport, roadway transport and marine transport. Air samples of benzene, toluene, ethylbenzene and m/p/o-xylene were collected by canisters and analysed by gas chromatography and mass selective detector technique. Results indicated that the in-vehicle VOC exposure levels were greatly influenced by the mode of transport. For benzene, mean concentration ranged from 4.8 to 6.1 microg x m(-3) in roadway transports, 3.0-3.8 microg x m(-3) in railway transports and it was 2.1 microg x m(-3) in ferry. Regardless of the results in MTR and air-conditioned buses, the VOC levels in roadway transport were the highest and was followed by railway transport. The exposure levels in marine transport were the lowest. The TEX concentrations were found to be substantially higher in air-conditioned buses and MTR trains than in other transports, suggesting the existence of additional solvent-related sources in their vehicle interiors. Measurements in non-air conditioned double deck vehicles indicate that there was slightly higher VOC levels in the lower deck than in the upper deck microenvironment. The average upper to lower deck exposure ratio ranged from 0.79 to 0.87 in trams and 0.78 0.83 in non-air-conditioned buses, depending on the compound of concern. The VOC exposure levels of public transport commuters in Hong Kong are far lower than those in most oversea cities. The experimental concentrations obtained in this study are within the relevant health-protecting limits as stated in the Hong Kong Indoor Air Quality Objective. Influences of recent VOC pollution control measures and local traffic characteristics on in-vehicle level are discussed. PMID- 12738210 TI - PCBs in the atmosphere of King George Island, Antarctica. AB - Polychlorinated biphenyls (PCBs) are ubiquitous anthropogenic compounds found even in the polar regions, mainly due to long-range atmospheric transport. In order to assess the occurrence of atmospheric PCBs in the vicinity of the Brazilian Antarctic Research Station (62 degrees 05'S, 58 degrees 23'W), a sampling survey was undertaken in the austral summer of 1995-1996. The concentrations of the PCB congeners in the air ranged from not detected to 33.2 pgm(-3). The lower chlorinated congeners (PCB-101 and below), which are more volatile and subject to long-range transport, predominated in the air samples and represented 66.7% of the total PCB concentrations. Atmospheric levels of PCBs were correlated with meteorological conditions and the highest levels were associated with the passage of the frontal systems coming from South America. PMID- 12738209 TI - Characterization of particulate, metallic elements of TSP, PM(2.5) and PM(2.5-10) aerosols at a farm sampling site in Taiwan, Taichung. AB - Atmospheric aerosol particles and metallic concentrations were monitored at the Experimental Farm of Tunghai University (EFTU) sampling site in this study. Total suspended particulate matter (TSP) was collected by using a PS-1 sampler at the farm-sampling site, in central Taiwan, from July 2001 to April 2002. At the same time, PM(2.5) and PM(2.5-10) were also measured with a Universal sampler from January 2002 to April 2002. Only subjects with the most complete data records on TSP sampling (N=43) and PM(10) sampling (N=23) were used in this analysis. Taichung Industrial Park, Taichung Kang Road (traffic) and a Hospital Incinerator surround the Experimental Farm of Tunghai University. Atmospheric concentrations of metallic elements were analyzed by a flame atomic absorption spectrophotometer (AA-680/G). The results indicated that the metallic elements Mg, Cu and Mn were the largest components in the TSP fraction; the metallic elements Fe and Cd were the largest composition in the PM(2.5-10) fraction; however, the metallic elements Pb, Zn, Cr and Ni were the largest abundance in the PM(2.5) fraction. The atmospheric metallic elements in the TSP, PM(2.5) and PM(2.5-10) fractions came different emission sources, such as soil, traffic, industry and resuspended particles. PMID- 12738211 TI - Experimental studies about the impact of traction sand on urban road dust composition. AB - Traffic causes enhanced PM(10) resuspension especially during spring in the US, Japan, Norway, Sweden and Finland, among other countries. The springtime PM(10) consists primarily of mineral matter from tyre-induced paved road surface wear and traction sand. In some countries, the majority of vehicles are equipped with studded tyres to enhance traction, which additionally increases road surface wear. Because the traction sand and the mineral matter from the pavement aggregate can have a similar mineralogical composition, it has been difficult to determine the source of the mineral fraction in the PM(10). In this study, homogenous traction sand and pavement aggregate with different mineralogical compositions were chosen to determine the sources of PM(10) particles by single particle analysis (SEM/EDX). This study was conducted in a test facility, which made it possible to rule out dust contributions from other sources. The ambient PM(10) concentrations were higher when traction sand was used, regardless of whether the tyres were studded or not. Surprisingly, the use of traction sand greatly increased the number of the particles originating from the pavement. It was concluded that sand must contribute to pavement wear. This phenomenon is called the sandpaper effect. An understanding of this is important to reduce harmful effects of springtime road dust in practical winter maintenance of urban roads PMID- 12738212 TI - Total suspended particulate manganese in ambient air in Montreal 1981-2000. AB - Our research group previously established the time variation of atmospheric Mn in Montreal for the period 1981-1992. The results indicated stable Mn concentrations between 1981 and 1990 followed by a decrease of almost 50% attributed to the closing of a ferromanganese plant located approximately 25 km from Montreal. The aim of this study is to assess the atmospheric concentrations of Mn in Montreal between 1993 and 2000, to compare these data to those for the period 1981-1992, and to evaluate the presence of any trend. Three sampling stations belonging to the Montreal Urban Community (MUC) air quality surveillance network were selected. Filters from 1993 to 2000 were obtained from the MUC archives. The first sample of each month was selected and chemical analysis was performed by neutron activation analysis. The mean annual atmospheric Mn concentrations in Montreal from 1993 to 2000 were found to be very stable. The mean Mn concentration calculated for this time period was 0.01 microg/m(3) for station 99 (characterized by a low traffic density), which was significantly different (P<0.05) from the mean of 0.03 microg/m(3) for stations 13 and 68 which had higher traffic densities. Even though Mn represented a small percentage of the TSP (varying between 0.02% and 0.14%), the comparison between Mn and TSP is interesting since the stable temporal profile of Mn since 1993 contrasts with the continuously decreasing atmospheric TSP concentrations. This observation suggests that the combustion of MMT used in gasoline could be contributing to maintaining stable atmospheric Mn levels. PMID- 12738213 TI - Role of quantitative mineralogical analysis in the investigation of sites contaminated by chromite ore processing residue. AB - A range of techniques, normally associated with mineralogical studies of soils and sediments, has been used to characterise the solid materials found on sites contaminated with chromite ore processing residue (COPR). The results show that a wide range of minerals are present, many of which are found extensively in high temperature synthetic systems such as cements and clinkers and their low temperature hydration products. Thus, the minerals in COPR can be divided into three main categories: unreacted feedstock ore (chromite); high temperature phases produced during chromium extraction (brownmillerite, periclase and larnite); and finally, minerals formed under ambient weathering conditions on the disposal sites (brucite, calcite, aragonite, ettringite, hydrocalumite, hydrogarnet). Apart from chromite, chromium occurs in brownmillerite, ettringite, hydrocalumite and hydrogarnet. Detailed study of the chemistry and stoichiometry of chromium-bearing phases in conjunction with phase abundance provides a quantitative description of the solid state speciation of Cr(III) and Cr(VI) in and amongst these minerals and in the COPR as a whole. Of the total chromium present in the samples most, approximately 60-70% is present as Cr(III) in chromite, whilst brownmillerite also represents a significant reservoir of Cr(III) which is approximately 15% of the total. The remaining chromium, between 20 and 25%, is present as Cr(VI) and resides mainly in hydrogarnet, and to a slightly lesser extent in hydrocalumite. In the latter, it is present principally in an exchangeable anionic form. Chromium (VI) is also present in ettringite, but quantitatively ettringite is a much less important reservoir of Cr(VI), accounting for approximately 3% of total chromium in one sample, but less than 1% in the other two. This description provides insight into the processes likely to control the retention and release of Cr(VI) from COPR-contaminated sites. Such information is of particular value in chemical modelling of the system, in risk assessment and in the development of methods of informed remediation. PMID- 12738214 TI - Incorporation of (15)N-TNT transformation products into humifying plant organic matter as revealed by one- and two-dimensional solid state NMR spectroscopy. AB - Solid-state double cross polarization magic angle spinning (DCPMAS) 15N 13C nuclear magnetic resonance (NMR) spectroscopy was applied to study the incorporation of TNT transformation products into humifying plant organic matter. For this approach, 13C-enriched plant material (Lolium perenne) was mixed with quartz sand and aerobically incubated for 11 months after addition of 15N(3) 2,4,6-trinitrotoluene (TNT). After successive extraction of the incubate with water, methanol and ethyl acetate, approximately 60% of the 15N added as 15N(3) TNT (15N(add)) remained in the solid organic residue (SOR-fraction). The acid insoluble fraction (AI) obtained after NaOH and HCl extractions contained approximately 20% of 15N(add). For both fractions, 15N NMR spectroscopy revealed an almost complete reduction of the TNT after 11 months of aerobic incubation. Most of the reduced nitrogen groups underwent further condensation. The corresponding DCPMAS NMR spectra allowed the identification of amides that are further substituted by alkyl groups that resist even acid hydrolysis. This assigns them to relatively stable compounds rather than to newly synthesized microbial peptides. The results of this study suggest further that the covalent binding of TNT transformation products to plant derived organic matter is mediated by alkylation and acetylation reactions, rather than by 1,4 addition of TNT-derived nitrogenous groups to quinones of the humic material. PMID- 12738215 TI - Biomarker responses and PAH uptake in Mya truncata following exposure to oil contaminated sediment in an Arctic fjord (Svalbard). AB - Expanding industrial activity (notably oil and gas exploration) in the Arctic requires assessment of the potential impact of chemicals on marine organisms living in seawater at low temperature. The bivalve Mya truncata is common in Svalbard fjord (Norway) where it experiences low temperature throughout the year. To measure the impact of polycyclic aromatic hydrocarbons (PAH) on M. truncata, the responses of three biomarkers [total oxyradical scavenging capacity-assay (TOSC), plasma membrane stability of haemocytes and respiration rates] were investigated from bivalves exposed to sediment contaminated with a PAH mixture (crude oil). After two weeks of exposure to the contaminated sediment, TOSC showed no change. The high TOSC value (4010+/-1339 unit mg(-1) protein) of Mya truncata (control group) is thought to protect biomolecules with a low turnover rate efficiently in a low food availability environment. In the exposed bivalves, the haemocyte cellular membranes were significantly destabilised compared with controls (P<0.05). Respiration rate of control and PAH-exposed individuals (0.055+/-0.020 mg O(2) dw(-1) h(-1)) was similar and relatively low as is typical for polar bivalves, reflecting a strategy to minimise energy expenditure to cope with 9 months of starvation. Bioaccumulation of PAH by M. truncata was also low, due probably to a combination of low metabolic rate and reduced solubility of the oil compounds at low temperature. Data indicated an uptake of mainly low molecular weight compounds (two and three ring molecules). A good correlation of logBAF(lipid) (bioaccumulation factor) and logK(ow) (octanol/water partitioning coefficient) was shown (r(2)=0.87). Tissue sensitivity and/or functional differences (digestive gland vs. haemocytes), PAH uptake route (dietary vs. gills), the low metabolic rate of M. truncata and the low environmental temperature (reducing the bioavailability of PAH) are factors that help explain these findings. PMID- 12738217 TI - Trace metal determination in total atmospheric deposition in rural and urban areas. AB - The wet, dry and total atmospheric depositions of some metals (Al, Cd, Cr, Cu, Fe, Na, Pb and Zn) were sampled at two sites and atmospheric fallout fluxes were determined for these locations. This work, led by two different research groups, allowed to reach two main goals: to define a simple analytical procedure to secure accurate shipboard sampling and analysis of atmospheric deposition, and to assess anthropogenic impacts of heavy metals to the environment. The first step about the validation step showed that the prevalent deposition type was dry deposition which represents 40, 60 and 80% for Cd, Cu and Pb, respectively. This prevalence of dry deposition in total atmospheric fallout supported the necessity of funnel wall rinsing which contains 30, 50 and 40% of collected Cd, Cu and Pb, respectively. Moreover, the reproducibility of atmospheric deposition collection was determined. The second step was performed by comparing two sampling sites. A rural sampling site, situated in Morvan's regional park (250 km south-east of Paris), was chosen for its isolation from any local and regional contamination sources. Fluxes obtained in this area were compared with those obtained at an urban site (Creteil, suburb of Paris) allowing comparison between urban and rural areas and demonstrating the impact of anthropogenic activities on atmospheric deposition of Cr, Cu and Pb. PMID- 12738216 TI - Distribution of platinum group elements and other traffic related elements among different plants along some highways in Germany. AB - Using ICP-MS and ICP-AES platinum group elements (Pt, Pd, Rh, Ru and Ir) and Ce, La, Nd, Pb and Zr have been determined in street dust, Taraxacum officinale (dandelion), Plantago lanceolata (plantain), Lolium multiflorum (annual ryegrass), Rhytidiadelphus squarrosus (moss) and Vascellum pratense (mushrooms) collected along highways and streets in Germany during 1999. Among the plants Taraxacum officinale (dandelion) reflects most adequately the pollution with the investigated elements matching the results from street dust. A strong positive correlation between all elements determined in the plants is established. Transfer factor for Pt between soil and plants has been determined in an agricultural experiment ranging between 0.004 and 0.008 for two types of soils. PMID- 12738218 TI - PAH emission from the open burning of agricultural debris. AB - The procedure for and results of a test study of polycyclic aromatic hydrocarbon (PAH) emission from the open burning of agricultural debris are discussed. For the investigation of open burning its simulation with emission sampling and analysis were applied. Further investigations also included observations of real instances of the open burning of wastes and collection of data on the structure and volume of combustible material. It was concluded that the open burning of agricultural wastes resulted in significant amounts of PAHs, but mainly the low molecular weight species (naphthalene, acenaphthylene, phenanthrene, fluoranthene, etc.). PMID- 12738219 TI - Marine mammals from northeast Atlantic: relationship between their trophic status as determined by delta13C and delta15N measurements and their trace metal concentrations. AB - The relationship between trophic position through delta13C and delta15N and trace metal concentrations (Zn, Cd, Cu and Hg) was investigated in the tissues of six marine mammal species from the Northeast Atlantic: striped dolphin Stenella coeruleoalba, common dolphin, Delphinus delphis, Atlantic white-sided dolphin Lagenorhynchus acutus, harbour porpoise Phocoena phocoena, white beaked-dolphin Lagenorhynchus albirostris, grey seal Halichoerus grypus stranded on French Channel and Irish coasts. White-beaked dolphins, harbour porpoises, white-sided dolphins, common and striped dolphins display the same relative and decreasing trophic position, as measured by delta15N values, along both the Irish and French channel coasts, reflecting conservative trophic habits between these two places. Hepatic and renal Cd concentrations were significantly correlated to muscle delta13C and delta15N values while Hg, Zn and Cu did not. These results suggest that Cd accumulation is partly linked to the diet while other factors such as age or body condition might explain Hg, Zn or Cu variability in marine mammals. Combined stable isotope and trace metal analyses appear to be useful tools for the study of marine mammal ecology. PMID- 12738220 TI - Vegetable oil spills on salt marsh sediments; comparison between sunflower and linseed oils. AB - The effects of a simulated spill of sunflower oil in salt marsh sediments were compared with an experiment with linseed oil. Sunflower and linseed oil penetrated the sediments at the same rates but different adsorption of the oils onto sediment particles resulted in the establishment of anaerobic conditions at shallower depths in sediments contaminated with linseed oil than with sunflower oil. The total lipid content of sunflower oil contaminated sediments remained almost stable for 6 months, whilst only 40% of linseed oil remained in the sediment after 2 months. Numbers of culturable heterotrophic bacteria and aerobic oil degrading bacteria in muddy sediment increased rapidly in response to the presence of the oils but bacterial numbers in sandy sediments increased more slowly for sunflower oil. Changes in fatty acid composition indicate similar degradation pathways for both oils but sunflower oil degraded more slowly than linseed oil and thus has the potential for longer lasting effects in marine environments. PMID- 12738221 TI - Temporal variations in dimethylsulphoniopropionate and dimethyl sulphide in the Zuari estuary, Goa (India). AB - Despite tropical estuarine systems representing important sites for active biogeochemical processes, studies on dimethyl sulphide (DMS) in these systems are sparse. Here we report on DMS and dimethylsulphoniopropionate (DMSP) variability in relation to physicochemical and biological parameters for a period of 14 months in a tropical estuarine environment. DMS and DMSP showed high temporal variations with maximal concentrations during the southwest monsoon coinciding with a dinoflagellate bloom. Dinoflagellates appear to be the major contributors to the DMSP pool. Average DMS and DMSP concentrations (surface and bottom) suggested that much of the DMSP produced is converted to forms other than DMS. Surface DMS varied between 0.3 and 15.4 nmol dm(-3) while DMSP ranged from 0.8 to 419.5 nmol dm(-3). The DMS flux was 0.03-1.9 microM m(-2) d(-1) (average=0.6 microM m(-2) d(-1)) during the study period, that concurs well with the values reported for temperate estuaries. PMID- 12738222 TI - Land inputs, behaviour and contamination levels of copper in a ria estuary (NW Spain). AB - Copper concentrations were measured in the sediments and dissolved phase of the water column in the Lerez Estuary (Pontevedra Ria, Galicia), NW Spain, and in the freshwater-sewage inputs to the estuary. Dissolved copper fluxes in the estuary were quantified. Results show that the freshwater end-member in the Lerez Estuary has minimal copper contamination ([Cu](Dissolved)=7+/-4 nM, Sediment Enrichment Factor=1) and the source of dissolved copper inputs is localized in the saline end-member ([Cu](Dissolved)=20-55 nM, Sediment Enrichment Factor=6-7). A non conservative behaviour of dissolved copper showing a net addition during estuarine mixing was observed. The budgetary calculations and the copper sediment distribution suggest that the major copper input to the estuary-ria system is located at the Marin-Placeres transect, playing an important role in the net addition of copper within the estuarine mixing. The importance of upwelling in the renewal of the Galician coastal waters and dilution of continental inputs is discussed. PMID- 12738223 TI - Growth of estuarine fish is associated with the combined concentration of sediment contaminants and shows no adaptation or acclimation to past conditions. AB - We tested whether the growth rates of small benthic fish (Gillichthys mirabilis) in three southern California estuaries corresponded with the local concentrations of contaminants. Fish originating from each estuary were transplanted to cages in each estuary in two reciprocal transplant experiments. The growth rates of caged fish, and the size-distribution of natural populations, showed the same pattern of difference among estuaries. Twelve metals and organic contaminants occurred in bulk sediments at concentrations close to their individual ERL values, and a simple index of their combined concentration (the mean ERL quotient) was inversely correlated to the growth of caged fish. Metals in the water column occurred at lower concentrations, relative to toxicity thresholds, than those in sediments and were unrelated to fish growth. Fish used in the field caging experiments, and other fish held in the laboratory under constant conditions, showed no difference in growth according to their estuary of origin. Fish originating from different estuaries also showed no consistent differences in their tissue burden of organic contaminants. Our results thus suggested no genetic adaptation or physiological acclimation to the past contaminant regime, but revealed a possible association between fish growth rates and the combined concentration of multiple sediment contaminants. PMID- 12738224 TI - L-type Ca2+ channels in atrial fibrillation: wallflowers or a vanishing act. PMID- 12738225 TI - Identity and function of cardiac K(ATP) channels. PMID- 12738226 TI - The L-type Ca2+-channel subunits alpha1C and beta2 are not downregulated in atrial myocardium of patients with chronic atrial fibrillation. AB - OBJECTIVE: Electrical remodeling as well as atrial contractile dysfunction after the conversion of atrial fibrillation (AF) to sinus rhythm (SR) are mainly caused by a reduction of the inward L-type Ca(2+) current (I(CaL)). We investigated whether the expression of L-type Ca2+-channel subunits was reduced in atrial myocardium of AF patients. METHODS: Right atrial appendages were obtained from patients undergoing coronary artery bypass graft surgery (CAD, n = 35) or mitral valve surgery (MVD, n = 37). Seventeen of the CAD patients and 18 of the MVD patients were in chronic (>3 months) AF, whereas the others were in SR. The protein expression of the L-type Ca2+-channel subunits alpha1C and beta2 was quantified by western blot analysis. Furthermore, we measured the density of dihydropyridine (DHP)-binding sites of the L-type Ca2+ channel using 3H-PN220-100 as radioligand. RESULTS: Surprisingly, the alpha1C and the beta2-subunit expression was not altered in atrial myocardium of AF patients. Also, the DHP binding site density was unchanged. CONCLUSION: The protein expression of the L type Ca2+-channel subunits alpha1C or beta2 is not reduced in atrial myocardium of AF patients. Therefore, the reduced I(CaL) might be due to downregulation of other accessory subunits (alpha2delta), expression of aberrant subunits, changes in channel trafficking or alterations in channel function. PMID- 12738227 TI - Distribution of Kir6.0 and SUR2 ATP-sensitive potassium channel subunits in isolated ventricular myocytes. AB - The subcellular distribution of ATP-sensitive potassium (K(ATP)) channel subunits in rat-isolated ventricular myocytes was investigated using a panel of subunit specific antisera. Kir6.1 subunits were associated predominantly with myofibril structures and were co-localized with the mitochondrial marker MitoFluor red (correlation coefficient (cc) = 0.63 +/- 0.05). Anti-Kir6.1 antibodies specifically recognized a polypeptide of 48 kDa in mitochondrial membrane fractions consistent with the presence of Kir6.1 subunits in this organelle. Both Kir6.2 and SUR2A subunits were distributed universally over the sarcolemma. Lower intensity antibody-associated immunofluorescence was detected intracellularly, which was correlated with the distribution of MitoFluor red in both cases (cc, Kir6.2, 0.56 +/- 0.05; SUR2A, 0.61 +/- 0.06). A polypeptide of 40 kDa was recognized by anti-Kir6.2-subunit antibodies in western blots of both microsomal and mitochondrial membrane fractions consistent with the presence of this subunit in the sarcolemma and mitochondria. Similarly, SUR2A and SUR2B subunits were detected in western blots of microsomal membrane proteins consistent with a sarcolemmal localization for these polypeptides. SUR2B subunits were shown in confocal microscopy to co-localize strongly with t-tubules (cc, 0.81 +/- 0.05). Together, the results indicate that Kir6.2 and SUR2A subunits predominate in the sarcolemma of ventricular myocytes consistent with a Kir6.2/SUR2A-subunit combination in the sarcolemmal K(ATP)channel. Kir6.1, Kir6.2 and SUR2A subunits were demonstrated in mitochondria. Combinations of these subunits would not explain the reported pharmacology of the mitochondrial K(ATP) channel (Mol Pharmacol 59 (2001) 225) suggesting the possibility of further unidentified components of this channel. PMID- 12738228 TI - Basal and alpha1-adrenergic-induced activity of minimal rat betaMHC promoters in cardiac myocytes requires multiple TEF-1 but not NFAT binding sites. AB - A culture model for cardiac hypertrophy, stimulation of neonatal rat cardiac myocytes by alpha(1)-adrenergic agonists, has been used extensively to identify transcription factors that activate genes during cardiac hypertrophy, such as skeletal alpha-actin, beta-myosin heavy chain (betaMHC), and B-natriuretic peptide. We used this culture model to further investigate transcription factors regulating the betaMHC promoter in cardiac myocytes under basal conditions and during hypertrophy. We found that the rat betaMHC promoter contains two other MCAT sites, in addition to the two MCATs reported previously. The four MCAT sites are conserved in some but not all of the mammalian betaMHC promoters examined, and all bind TEF-1 but with varying affinity. As assayed by transient transfection into cardiac myocytes, the four MCATs within 348 bp of the transcription start site are required for full activity of the rat betaMHC promoter in the absence and presence of the alpha(1)-adrenergic agonist phenylephrine (PE). We found that the betaMHC promoter also contains a binding site for the NFAT family of transcription factors, which are activated by calcineurin and are implicated in the hypertrophic process. Although this site bound NFAT3 in vitro and has been reported to be required for betaMHC promoter activity in slow skeletal muscle, mutation of the site had no effect on basal or on PE-induced activity of the promoter in cardiac myocytes. Our results show that full activity of minimal betaMHC promoters in the presence and absence of hypertrophic agents requires multiple MCAT sites but not NFAT-binding sites. PMID- 12738229 TI - Regulation of the PAI-1 promoter by circadian clock components: differential activation by BMAL1 and BMAL2. AB - Circadian variation in plasminogen activator inhibitor-1 (PAI-1) production likely contributes to increased risk of myocardial infarction and decreased efficacy of thrombolytic therapy during the morning. In this study, we characterize the abilities of fundamental molecular components of intrinsic circadian clocks to regulate the human PAI-1 promoter in transfected endothelial cells. Both CLOCK:BMAL1 and CLOCK:BMAL2 heterodimers activate the PAI-1 promoter through requisite proximal (-565 to -560 bp) and distal (-680 to -675 bp) E-box enhancers. Although the distal E-box overlaps the 4G/5G polymorphism of the PAI-1 promoter, allelic variation at this site does not influence CLOCK:BMAL1-and CLOCK:BMAL2-mediated transactivation. Together, CLOCK:BMAL1 and CLOCK:BMAL2 make additive contributions to PAI-1 gene transcription. While the abilities of these heterodimers to activate gene expression differ by twofold, the susceptibilities of these circadian activators to inhibition by period and cryptochrome proteins are equivalent and redox independent. Given that BMAL1 and BMAL2 differ in their spatiotemporal distributions, such distinctions may allow intrinsic circadian clocks to modulate the amplitudes of their oscillators, while maintaining circadian periodicity. In this way, fundamental circadian clock components may drive circadian variation in PAI-1, which in turn influences the pathogenesis, timing, and treatment of acute atherothrombotic events. PMID- 12738231 TI - The role of arginine vasopressin and its receptors in the normal and failing rat heart. AB - OBJECTIVE: To evaluate the role of arginine vasopressin (AVP) in the normal and post-myocardial infarction (post-MI) hearts, and to investigate whether chronic AVP receptor antagonism can attenuate post-infarct ventricular remodeling. BACKGROUND: A number of neurohormones, like norepinephrine and angiotensin II, have detrimental effects in heart failure (HF) and inhibiting them is beneficial. AVP shares some important properties with these hormones and is activated in HF. However, its role in the syndrome of HF, especially the effect of AVP inhibition, is largely undefined. METHODS AND RESULTS: Effects of AVP-V1a and AVP-V2 receptor stimulation on normal rat cardiomyocyte contractile function, intracellular calcium [Ca2+]i, inositol 3 phosphate (IP3) generation and cell survival were studied. In post-MI rats, AVP receptor function was assessed in cardiomyocytes as well as isolated working hearts. AVP receptor total number (ligand binding) and mRNA levels (RT-PCR) were measured in myocytes. Finally, the effects of chronic AVP-V1a blockade were assessed in post-MI rats using echo and morphometry to measure ventricular remodeling. Normal cardiomyocytes showed a dose-dependent increase in myocyte contractile function, [Ca2+]i, and IP3 generation in response to AVP-V1a receptor stimulation. AVP-V2 agonists had no effect. Cells from MI hearts showed reduced inotropic response to AVP-V1a stimulation. Myocyte AVP-V1a receptor number and receptor mRNA were decreased. Prolonged exposure to AVP reduced cellular viability. However, chronic AVP-V1a blockade did not attenuate structural remodeling or improve function in post-MI hearts. CONCLUSIONS: Isolated adult rat cardiomyocytes bear AVP-V1a receptors that mediate inotropic effects through the IP3 pathway. These receptors are functionally and numerically downregulated in the post-MI remodeled hearts. Despite evidence that AVP is cytotoxic to cells, chronic AVP-V1a receptor blockade does not attenuate post-MI ventricular remodeling in this model. Thus, AVP may not play a major role in the structural progression of HF. PMID- 12738232 TI - Differential accumulation of non-transferrin-bound iron by cardiac myocytes and fibroblasts. AB - Cardiac myocytes accumulate iron preferentially over fibroblast-like non myocytes, both in clinical iron overload and when the cells are grown together in culture. In order to determine whether this reflects the tissue context or is an inherent property of the cells, we studied iron transporters, transport kinetics, and iron efflux in homogeneous cultures of rat cardiac myocytes and fibroblasts. In both cells, the rate of uptake of 59Fe from transferrin was insignificant, compared to the rate of uptake from non-transferrin-bound iron (NTBI). Expression of transferrin receptor mRNA and protein, and divalent metal transporter 1 (DMT1) mRNA, could not account for any difference in iron accumulation, and proportional efflux after iron loading was similar in both cells. Nevertheless, iron accumulation from NTBI over 72 h was greater in myocytes as determined by histological staining and quantitative iron measurement. NTBI uptake was greater for Fe2+ than Fe3+ in both cells, was increased by iron loading in both cells to a similar extent, and was characterized bysimilar Michaelis constants (K(m)) in all cases (redox state and presence or absence of iron loading). However, V(max) values were about 10-fold higher in myocytes. We conclude that preferential iron accumulation in cardiac myocytes, compared to fibroblasts, is due to a higher capacity of the NTBI-transporter system, and reflects an inherent difference in NTBI acquisition by the individual cell types. PMID- 12738230 TI - Beta-adrenergic receptor blockade modulates Bcl-X(S) expression and reduces apoptosis in failing myocardium. AB - The mechanisms by which beta-adrenergic receptor (beta-AR) blockade modulates apoptosis in heart failure (HF) are unclear. We examined the impact of beta-AR blockade with metoprolol on myocardial remodeling, apoptosis, pro-apoptotic (Fas, Fas ligand, Bax, and Bcl-X(S)) and anti-apoptotic (Bcl-X(L)and Bcl-2) gene expression, and Bcl-X(L) and Bcl-X(S) protein in post-infarction HF in rats. In untreated rats, there was significant (P < 0.001) LV dilatation and systolic dysfunction compared to sham. Myocardial apoptosis was significantly increased (P < 0.005). Fas, Bax, and Bcl-2 mRNA expression was unchanged. However, Fas ligand mRNA and Bcl-X(S) mRNA and protein, all undetectable in sham, were markedly elevated (P < 0.001), whereas Bcl-X(L) mRNA and protein was unchanged. Immunohistochemistry confirmed increased Bcl-X(S) staining in failing myocardium, with unchanged Bcl-X(L). Metoprolol treatment resulted in: (1) improved LV remodeling (P < 0.025), (2) reduced myocardial apoptosis (P < 0.005), and (3) selective reduction in myocardial Bcl-X(S) expression (P < 0.001) without change in Fas, Fas ligand, Bax, Bcl-2, or Bcl-X(L). Studies in isolated rat myocytes revealed that prolonged isoproterenol (ISO) stimulation significantly increased Bcl-X(S) protein, reducing the Bcl-X(L)/X(S) ratio and myocyte survival (P < 0.005). ISO-induced Bcl-X(S) expression was significantly attenuated (P < 0.001) by both metoprolol and CGP20712A, a beta1-AR selective antagonist, but not by ICI118,551, a beta2-AR selective antagonist. We conclude that adrenergic activation, such as occurs in HF, increases pro-apoptotic Bcl-X(S) expression via the beta1-AR. beta-AR blockade in HF reduces myocardial apoptosis; attenuation of Bcl-X(S) expression may be one mechanism underlying this effect. PMID- 12738233 TI - T-type Ca2+ current contribution to Ca2+-induced Ca2+ release in developing myocardium. AB - In normal adult-ventricular myocardium, Ca2+-induced Ca2+ release (CICR) from the sarcoplasmic reticulum (SR) is activated via Ca2+ entry through L-type Ca2+ channels. However, embryonic-ventricular myocytes have a prominent T-type Ca2+ current (ICa,T). In this study, the contribution of ICa,T to CICR was determined in chick-ventricular development. Electrically stimulated Ca2+ transients were examined in myocytes loaded with fura-2 and Ca2+ currents with perforated patch clamp. The results show that the magnitudes of the Ca2+ transient, L-type Ca2+ current (ICa,L) and ICa,T, decline with development with the majority of the decline of transients and ICa,L occurring between embryonic day (ED) 5 and 11. Compared to controls, the magnitude of the Ca2+ transient in the presence of nifedipine was reduced by 41% at ED5, 77% at ED11, and 78% at ED15. These results demonstrated that the overall contribution of ICa,T to the transient was greatest at ED5, while ICa,L was predominate at ED11 and 15. This indicated a decline in the contribution of ICa,T to the Ca2+ transient with development. Nifedipine plus caffeine was added to deplete the SR of Ca2+ and eliminate the occurrence of CICR due to ICa,T. Under these conditions, the transients were further reduced at all three developmental ages, which indicated that a portion of the Ca2+ transients present after just nifedipine addition was due to CICR stimulated by ICa,T. These results indicate that Ca2+ entry via T-type channels plays a significant role in excitation-contraction coupling in the developing heart that includes stimulation of CICR. PMID- 12738235 TI - Matrix metalloproteinase abundance in human myocardial fibroblasts: effects of sustained pharmacologic matrix metalloproteinase inhibition. AB - BACKGROUND: A cause-effect relationship has been established between matrix metalloproteinases (MMPs) and left ventricular (LV) myocardial remodeling through the use of pharmacologic MMP inhibitors. However, the direct effects of MMP inhibition on MMPs and endogenous tissue inhibitors of metalloproteinases (TIMPs) in LV human myocardial fibroblasts (LVHMFs) remain unknown. This study measured MMP-2, MMP-9, MMP-13, MT1-MMP, and TIMP-1 release in LVHMFs. METHODS AND RESULTS: LVHMF cultures were established from six individual patients (passages 2-5) and incubated with and without the broad-spectrum MMP inhibitor PD166793 (100 microM) for 12-36 h. While PD166793 did not influence MMP-2 release, MMP-9 levels based on substrate zymography increased at 36 h by over 30% (P < 0.05). TIMP-1 levels increased in a time-dependent manner with no effect from PD166793 incubation. However, the MMP-9/TIMP-1 ratio was increased by over 20% from time-matched values following 12-36 h of exposure to PD166793 (P < 0.05). Similar results obtained after incubation of LVHMF cultures with the broad-spectrum MMP inhibitor Batimastat (BB-94) suggest that these observations are due to a general class effect of broad-spectrum MMP inhibitors. CONCLUSIONS: This study is the first to demonstrate that a selective induction and release of an MMP species occurs with sustained exposure to pharmacologic MMP inhibition in LVHMFs. These observations may have particular importance with respect to controlling this proteolytic system in the context of LV myocardial remodeling. PMID- 12738234 TI - Mechanism of cyclooxygenase-2 upregulation in late preconditioning. AB - Although the cardioprotection of late preconditioning (PC) is known to be mediated by both inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2), the signaling mechanism responsible for COX-2 upregulation and the interaction between iNOS and COX-2 remain unknown. A total of 122 mice were used to address this issue. In wild-type mice preconditioned with six cycles of 4-min coronary occlusion-4-min reperfusion, ischemic PC resulted in rapid activation of nuclear STAT1/3 through tyrosine phosphorylation (STAT1: 339 +/- 48% of control; STAT3: 389 +/- 46% of control) and increased STAT1/3-DNA binding activity (687 +/- 58% of control) at 30 min after PC, with subsequent upregulation of COX-2 protein (373 +/- 60% of control) and activity(increased myocardial levels of PGE2, PGF(2alpha), and 6-keto-PGF(1alpha)) at 24 h. However, COX-1 protein was not changed 24 h after ischemic PC. Pretreatment with the Janus tyrosine kinase (JAK) inhibitor AG-490 before the six occlusion-reperfusion cycles blocked both the tyrosine phosphorylation of STAT1/3 and the subsequent upregulation of COX-2 protein, demonstrating a necessary role of the JAK-STAT pathway in the induction of COX-2. Targeted disruption of the iNOS gene (iNOS-/-) did not block the increased expression of COX-2 protein 24 h after ischemic PC but completely blocked the increase in COX-2 activity, whereas targeted disruption of the COX-2 gene (COX-2-/-) did not alter ischemic PC-induced iNOS induction. Immunoprecipitation of preconditioned heart tissues with anti-COX-2 antibodies followed by immunoblotting with anti-iNOS antibodies revealed that the increased iNOS protein co-precipitated with COX-2. We conclude that (i) the upregulation of COX-2 protein expression after ischemic PC is mediated by a JAK1/2-STAT1/3 signaling cascade; (ii) COX-2 activity requires upregulated iNOS and iNOS-derived NO; and (iii) COX-2 forms complexes with iNOS, supporting a direct interaction between these two proteins. To our knowledge, this is the first evidence that myocardial COX-2 is upregulated via a JAK1/2-STAT1/3 pathway. PMID- 12738236 TI - Na+ channel mutation leading to loss of function and non-progressive cardiac conduction defects. AB - BACKGROUND: We previously described a Dutch family in which congenital cardiac conduction disorder has clinically been identified. The ECG of the index patient showed a first-degree AV block associated with extensive ventricular conduction delay. Sequencing of the SCN5A locus coding for the human cardiac Na+ channel revealed a single nucleotide deletion at position 5280, resulting in a frame shift in the sequence coding for the pore region of domain IV and a premature stop codon at the C-terminus. METHODS AND RESULTS: Wild type and mutant Na+ channel proteins were expressed in Xenopus laevis oocytes and in mammalian cells. Voltage clamp experiments demonstrated the presence of fast activating and inactivating inward currents in cells expressing the wild type channel alone or in combination with the beta1 subinut (SCN1B). In contrast, cells expressing the mutant channels did not show any activation of inward current with or without the beta1 subunit. Culturing transfected cells at 25 degrees C did not restore the Na+ channel activity of the mutant protein. Transient expression of WT and mutant Na+ channels in the form of GFP fusion proteins in COS-7 cells indicated protein expression in the cytosol. But in contrast to WT channels were not associated with the plasma membrane. CONCLUSIONS: The SCN5A/5280delG mutation results in the translation into non-function channel proteins that do not reach the plasma membrane. This could explain the cardiac conduction defects in patients carrying the mutation. PMID- 12738238 TI - The proteasome: structure, function, and role in the cell. AB - The proteasome is a multisubunit enzyme complex that plays a central role in the regulation of proteins that control cell-cycle progression and apoptosis, and has therefore become an important target for anticancer therapy. Before a protein is degraded, it is first flagged for destruction by the ubiquitin conjugation system, which ultimately results in the attachment of a polyubiquitin chain on the target protein. The proteasome's 19S regulatory cap binds the polyubiquitin chain, denatures the protein, and feeds the protein into the proteasome's proteolytic core. The proteolytic core is composed of 2 inner beta rings and 2 outer alpha rings. The 2 beta rings each contain 3 proteolytic sites named for their trypsin-like, post-glutamyl peptide hydrolase-like (PGPH) (i.e., caspase like), or chymotrypsin-like activity. Inhibition of the proteasome results in cell-cycle arrest and apoptosis. In in vitro and in vivo animal studies, inhibition of the proteasome via bortezomib (VELCADE; formerly, PS-341, LDP-341, and MLN341) had antitumor activity against numerous tumor types either alone or in combination with conventional chemotherapeutic agents; these results provided the rationale for a broad clinical trial program. Bortezomib is currently in phase III trials for myeloma and is in early clinical development for numerous other tumor types. PMID- 12738239 TI - The tumor microenvironment: focus on myeloma. AB - A wide variety of cellular responses that may afford tumor cells drug-tolerance characteristics. Overexpression of plasma membrane efflux pumps, up-regulation of anti-apoptosis factors, down-regulation of proapoptosis factors, subcellular redistribution of drug targets, and up-regulation of detoxifying enzymes are just a few known mechanisms of cancer cell resistance. In addition to these individual cell adaptations, cellular drug resistance also appears to be mediated by the binding of tumor cells to extracellular matrix (ECM) proteins. Cell adhesion mediated drug resistance (CAM-DR) is particularly relevant in hematologic malignancies such as multiple myeloma, where myeloma cells localize in the bone marrow and interact with stroma and stromal cells, initiating the production of proteins that stimulate or support tumor survival. Thus, CAM-DR provides a plausible explanation for the protective mechanisms associated with myeloma cell adhesion and demonstrates that the tumor microenvironment may hold the key to elucidating how tumor cells resist chemotherapy. PMID- 12738240 TI - Rationale for the treatment of solid tumors with the proteasome inhibitor bortezomib. AB - Given its role in cellular metabolism, the proteasome could prove to be a critical target that can be exploited in treating cancer. In preclinical studies, several mechanisms for bortezomib's activity in multiple myeloma cells have been identified (e.g., NF-kappaB inhibition); antitumor activity with bortezomib has been seen in myeloma patients, thereby supporting the validity of the preclinical work. Similar mechanisms may be in play in solid tumors, and cell culture and xenograft data suggest bortezomib may be active in a wide range of tumor types. One promising possibility is the use of bortezomib for the treatment of chemoresistant tumors. Chemoresistance can be caused by a number of cellular factors; NF-kappaB is a prominent instigator of chemoresistance, and proteasome inhibition was an effective means of preventing NF-kappaB activation in myeloma and several solid tumor laboratory studies. However, the inhibition of NF-kappaB may not be the only mechanism for antitumor activity. This review explores the use of proteasome inhibitors to subvert intrinsic resistance mechanisms, disrupt inducible chemoresistance, or augment the mechanisms of action of standard chemotherapeutics. Thus, in addition to providing another target for anticancer treatment, proteasome inhibition may also provide a means to treat refractory tumors. PMID- 12738241 TI - Clinical update: proteasome inhibitors in hematologic malignancies. AB - The proteasome inhibitor bortezomib (VELCADE; formerly PS-341, LDP-341, MLN341) is a novel dipeptide boronic acid. In cell culture and xenograft models, bortezomib showed potent activity, enhanced the sensitivity of cancer cells to traditional chemotherapeutics, and appeared to overcome drug resistance. In vitro, bortzomib downregulated the NF-kappaB pathway. NF-kappaB is a transcription factor that enhances the production of growth factors (e.g., IL-6), cell-adhesion molecules, and anti-apoptotic factors, all of which contribute to the growth of the tumor cell and/or protection from apoptosis. Phase II trials have been conducted in patients with relapsed and refractory multiple myeloma (SUMMIT trial, 202 patients) or relapsed myeloma (CREST trial, n=54) using a 1.3mg/m(2) dose given twice weekly for 2 weeks (days 1, 4, 8, 11; rest days 12 21). Both trials showed responses (including complete responses) with manageable toxicities, forming the basis for an ongoing phase III trial comparing response to bortezomib versus high-dose dexamethasone. PMID- 12738242 TI - Clinical update: proteasome inhibitors in solid tumors. AB - The proteasome plays a critical role in regulating the cell cycle, neoplastic growth, and metastasis. Bortezomib (VELCADE; formerly PS-341, LDP-341, MLN341) is a novel dipeptide boronic acid that is the first proteasome inhibitor to have progressed to clinical trials. Preclinical research has shown that through the prevention of IkappaB degradation, bortezomib may block chemotherapy-induced NF kappaB activation and augment the apoptotic response to chemotherapeutic agents. Bortezomib also appeared to increase the stabilization of p21 and p27, as well as transcription factor p53. In preclinical models of breast, lung, pancreatic, and ovarian tumor types, bortezomib inhibited tumor growth and demonstrated anti angiogenic properties. Bortezomib exhibited the greatest activity when combined with standard chemotherapeutic agents, such as irinotecan, gemcitabine, and docetaxel, suggesting its potential additive/syngeristic role in overcoming resistance to conventional chemotherapy. Preliminary data from early clinical trials suggest that bortezomib can be given at pharmacologically active doses in combination with standard doses of chemotherapy with manageable toxicities. Responses have been seen and no evidence of additive toxicity has been exhibited in combination agent trials. PMID- 12738243 TI - Therapeutic anti-cancer targets upstream of the proteasome. AB - Polyubiquitination of a protein is generally the first step in its degradation. This article discusses how altered protein destruction pathways impact the cell cycle and allow for abnormal cell proliferation, and explores how this process can be utilized in anticancer therapy. There are several levels of possible therapeutic intervention in ubiquitin-dependent proteolysis pathways upstream of the proteasome. In principle, targeting specific components of the ubiquitin system may offer an opportunity to develop selective drugs. However, the fact that general proteasome inhibitors have been demonstrated to be effective in cancer therapy suggests that other ubiquitin components that are common to many destruction pathways may also be clinically useful. We will, therefore, evaluate both the specific, rate-limiting enzymes and a number of general, nonselective enzymes as targets for anticancer therapy. Potential nonselective therapeutic strategies that are under investigation in a variety of human cancers include the identification and inhibition of individual F-box proteins, such as Skp2, and the inhibition of the ubiquitin ligases such as the SCF family, Mdm2, and Efp. A general pathway under investigation is the cullin neddylation and deneddylation system, with promising enzymatic targets such as csn5 and Rpn11. PMID- 12738244 TI - Teasing apart the complex genetics of human autoimmunity: lessons from rheumatoid arthritis. PMID- 12738245 TI - Environmental polycyclic aromatic hydrocarbons, benzo(a) pyrene (BaP) and BaP quinones, enhance IgE-mediated histamine release and IL-4 production in human basophils. AB - Polycyclic aromatic hydrocarbons (PAHs) are major components of diesel exhaust particles found in pollutant respirable particles. There is growing evidence that these fossil fuel combustion products exacerbate allergic inflammation. Basophils contribute to allergic inflammation through the release of preformed and granule derived mediators. To determine whether allergens and PAHs interact, we incubated human basophils with PAHs and measured the release of histamine and IL-4 with and without added antigen. None of the PAHs induced mediator release by itself and none affected total cellular histamine levels. However, several PAHs enhanced histamine release and IL-4 production in response to crosslinking the high affinity IgE receptor, Fc epsilon RI. The enhancement seen with 1,6-BaP-quinone involved an increase in tyrosine phosphorylation in several different substrates, including the Fc epsilon RI-associated tyrosine kinase, Lyn, and elevated reactive oxygen species (ROS) levels detected by dichlorofluorescein fluorescence and flow cytometry. The PAH-induced enhancement of mediator release and ROS production could be inhibited with the antioxidant N-acetylcysteine. These data provide further evidence that environmental pollutants can influence allergic inflammation through enhanced Fc epsilon RI-coupled mediator release from human basophils. PMID- 12738246 TI - Effects of multidose combination chemotherapy on the humoral immune system. AB - Patients receiving multidose combination chemotherapy are at risk for severe, life-threatening infections, caused by among others encapsulated bacteria like Streptococcus pneumoniae. The splenic marginal zone is essential in the initiation of immune responses to S. pneumoniae. We analyzed effects of multidose combination chemotherapy on B-cell subpopulations. Immune response capacity was evaluated by using Pneumovax (PPS) or Tetavax (TT) as antigenic challenge. Three days after finishing therapy, all B-cell subpopulations in bone marrow and spleen were severely reduced, including the mature marginal zone B-cell population. When analyzing the anti-PPS immune response capacity at 3 days after finishing therapy, we found that the IgM antibody levels did not differ significantly from control immunized rats. The IgG antibody levels were significantly lower compared to control immunized rats but still significantly higher compared to unimmunized rats. The depletion of marginal zone B cells by multidose combination chemotherapy most likely contributes to the prolonged period that patients are at risk for developing severe infections after chemotherapy, despite the capacity to generate sufficient antibody levels. It is conceivable that the local (temporary) loss of immunological memory, together with the supposed inability to generate a humoral response in a short time frame, plays an important role in this vulnerability. PMID- 12738247 TI - The immune modulator FYT720 prevents autoimmune diabetes in nonobese diabetic mice. AB - FTY720 is a novel immune regulatory drug derived from the fungal sphingosine analog ISP-1 (myriocin). FTY720 causes a redistribution of lymphocytes from circulation to secondary lymphoid tissues. Type 1 diabetes is an autoimmune disorder caused by cellular-mediated destruction of insulin-producing pancreatic beta cells in the islets of Langerhans. Indeed, local infiltration of islets by mononuclear cells is the hallmark of Type 1 diabetes. Based on both FTY720's action and the involvement of cellular infiltration in the disease progression, we tested FTY720 for its ability to prevent autoimmune diabetes in diabetes prone, nonobese diabetic (NOD) mice. We found that treatment with FTY720 completely prevented NOD mice from developing autoimmune diabetes. The FTY720 treated animals showed both reduced numbers of circulating lymphocytes and sharply diminished cellular infiltration of pancreatic islets. These results suggest that FTY720 may be effective in prevention of autoimmune diabetes or in slowing its progression. PMID- 12738248 TI - Determinants of HIV-specific CD8 T-cell responses in HIV-infected pediatric patients and enhancement of HIV-gag-specific responses with exogenous IL-15. AB - Cellular immune responses play a central role in controlling HIV-1 infection. HIV specific IFN-gamma production by CD8 T cells was evaluated in 17 HLA-A2+ HIV infected pediatric patients (age range 1 month to 16 years) in an ELISPOT assay. Most patients (15/17) exhibited responses to HIV-gag, followed by responses to envelope gp120, gp41, and V3 loop. Only 7 patients responded to all four antigenic peptides. Treatment-related immune reconstitution of CD4 T cells was associated with increase in gag-specific responses, but these declined with prolonged viral suppression. Exogenous IL-15 resulted in augmentation of HIV-gag specific response in 71% of patients, while IL-2 and IL-7 had variable effects, augmenting responses in 25% patients. Thus, HIV-specific CD8 T-cell responses are dependent on both CD4 T-cell help and antigenic stimulation. The cytokine IL-15 may be a useful modality as adjunctive therapy to augment HIV-specific memory CD8 T cells. PMID- 12738249 TI - The calcium-binding protein S100A12 induces neutrophil adhesion, migration, and release from bone marrow in mouse at concentrations similar to those found in human inflammatory arthritis. AB - We investigated the proinflammatory activities of S100A12 in the context of synovial inflammation. S100A12 levels were increased in the synovial fluids and plasma of patients with gout, rheumatoid arthritis, psoriatic arthritis, and undetectable in osteoarthritis, a noninflammatory disorder. S100A12 proved to induce neutrophil adhesion to fibrinogen via Mac-1 at concentrations similar to those found in the synovial fluids. Similar concentrations induced the recruitment of large numbers of neutrophils and monocytes in the murine air pouch model. To characterize the effect of increased S100A12 plasma levels, mice were injected intravenously with S100A12. This led to the mobilization of neutrophils from the bone marrow to the peripheral blood. These results suggest that S100A12 stimulates the accumulation of neutrophil by inducing their release from the bone marrow, as well as by activating their adhesion and migration toward inflammatory sites. PMID- 12738250 TI - The tandem-repeat polymorphism of the DC-SIGNR gene does not affect the susceptibility to HIV infection and the progression to AIDS. AB - DC-SIGNR is a C-type lectin that functions as a transreceptor for HIV-1. The exon 4 of the DC-SIGNR gene comprises a variable number of 69-bp tandem repeats, encoding for parts of the extracellular protein domain. Here, we analyzed the relevance of this gene polymorphism for the interindividual transmission of HIV-1 and the progression to AIDS. A cross-sectional comparison between HIV-1-infected patients (n = 391) and healthy volunteers (n = 134) did not reveal significant differences with regard to the DC-SIGNR allele distribution. Moreover, DC-SIGNR allele frequencies were similar in slowly progressing HIV patients (n = 31) and patients who rapidly progressed to AIDS (n = 46). Additionally, in a cohort of 149 newly HIV-infected patients, no relationship was found between HIV set point viremia and DC-SIGNR genotypes. Thus, the DC-SIGNR tandem-repeat polymorphism in exon 4 does not have a significant impact on the host's susceptibility to HIV and the clinical progression to AIDS. PMID- 12738251 TI - Blood and intragraft CD27 gene expression in cardiac transplant recipients. AB - The present study investigated gene expression of costimulatory molecule CD27 in relation to the occurrence of acute cardiac rejection. CD27 transcripts were measured by means of quantitative competitive reverse transcriptase-polymerase chain reaction in 120 endomyocardial biopsies and in 89 samples of blood mononuclear cells from 31 recipients. Higher levels of CD27 transcripts were observed in biopsies with rejection than in samples without rejection (medians, 7.1 and 1.9; P = 0.06). In contrast, blood mononuclear cells collected during rejection showed lower levels than blood mononuclear cells from rejection-free periods (medians, 3.3 vs. 7.9; P = 0.03). Considering only endomyocardial biopsies without rejection, the values were lower in samples from recipients who did not present any rejection during the first 6 months after transplantation than in those from recipients who had at least one rejection during the same period (medians, 0 vs. 3.5, P < 0.001; percentage of biopsies expressing CD27, 44% vs. 77%). In conclusion, the presence of intragraft CD27 mRNA may identify recipients at risk for developing acute rejection. PMID- 12738252 TI - Naturally produced organohalogens. Proceedings of the Second International Conference on Naturally Produced Organohalogens. Heidelberg, Germany. September 30-October 3, 2001. PMID- 12738253 TI - The diversity of naturally produced organohalogens. AB - More than 3800 organohalogen compounds, mainly containing chlorine or bromine but a few with iodine and fluorine, are produced by living organisms or are formed during natural abiogenic processes, such as volcanoes, forest fires, and other geothermal processes. The oceans are the single largest source of biogenic organohalogens, which are biosynthesized by myriad seaweeds, sponges, corals, tunicates, bacteria, and other marine life. Terrestrial plants, fungi, lichen, bacteria, insects, some higher animals, and even humans also account for a diverse collection of organohalogens. PMID- 12738254 TI - Biological dehalogenation and halogenation reactions. AB - A large number of halogenated compounds is produced by chemical synthesis. Some of these compounds are very toxic and cause enormous problems to human health and to the environment. Investigations on the degradation of halocompounds by microorganisms have led to the detection of various dehalogenating enzymes catalyzing the removal of halogen atoms under aerobic and anaerobic conditions involving different mechanisms. On the other hand, more than 3500 halocompounds are known to be produced biologically, some of them in great amounts. Until 1997, only haloperoxidases were thought to be responsible for incorporation of halogen atoms into organic compounds. However, recent investigations into the biosynthesis of halogenated metabolites by bacteria have shown that a novel type of halogenating enzymes, FADH(2)-dependent halogenases, are involved in biosyntheses of halogenated metabolites. In every gene cluster coding for the biosynthesis of a halogenated metabolite, isolated so far, one or several genes for FADH(2)-dependent halogenases have been identified. PMID- 12738255 TI - Pattern and sources of naturally produced organohalogens in the marine environment: biogenic formation of organohalogens. AB - The pattern of organohalogens found in the marine environment is complex and includes compounds, only assignable to natural (chloromethane) or anthropogenic (hexachlorobenzene, PCBs) sources as well as compounds of a mixed origin (trichloromethane, halogenated methyl phenyl ether).The chemistry of the formation of natural organohalogens is summarized. The focus is put on volatile compounds carrying the halogens Cl, Br, and I, respectively. Though marine natural organohalogens are quite numerous as defined components, they are mostly not produced as major compounds. The most relevant in terms of global annual production is chloromethane (methyl chloride). The global atmospheric mixing ratio requires an annual production of 3.5-5 million tons per year. The chemistry of the group of haloperoxidases is discussed. Incubation experiments reveal that a wide spectrum of unknown compounds is formed in side reactions by haloperoxidases in pathways not yet understood. PMID- 12738256 TI - The role of halogen species in the troposphere. AB - While the role of reactive halogen species (e.g. Cl, Br) in the destruction of the stratospheric ozone layer is well known, their role in the troposphere was investigated only since their destructive effect on boundary layer ozone after polar sunrise became obvious. During these 'Polar Tropospheric Ozone Hole' events O(3) is completely destroyed in the lowest approximately 1000 m of the atmosphere on areas of several million square kilometres. Up to now it was assumed that these events were confined to the polar regions during springtime. However, during the last few years significant amounts of BrO and Cl-atoms were also found outside the Arctic and Antarctic boundary layer. Recently even higher BrO mixing ratios (up to 176 ppt) were detected by optical absorption spectroscopy (DOAS) in the Dead Sea basin during summer. In addition, evidence is accumulating that BrO (at levels around 1-2 ppt) is also occurring in the free troposphere at all latitudes. In contrast to the stratosphere, where halogens are released from species, which are very long lived in the troposphere, likely sources of boundary layer Br and Cl are autocatalytic oxidation of sea salt halides (the 'Bromine Explosion'), while precursors of free tropospheric BrO and coastal IO probably are short-lived organo-halogen species. At the levels suggested by the available measurements reactive halogen species have a profound effect on tropospheric chemistry: In the polar boundary layer during 'halogen events' ozone is usually completely lost within hours or days. In the free troposphere the effective O(3) losses due to halogens could be comparable to the known photochemical O(3) destruction. Further interesting consequences include the increase of OH levels and (at low NO(X)) the decrease of the HO(2)/OH ratio in the free troposphere. PMID- 12738257 TI - Fluxes of trichloroacetic acid between atmosphere, biota, soil, and groundwater. AB - Trichloroacetic acid (TCA), in former times used as a herbicide in agriculture, is now ubiquitous and almost evenly distributed in precipitations of the Northern and Southern Hemisphere, despite larger emissions of the possible precursors tetrachloroethene and 1,1,1-trichloroethane in the Northern Hemisphere. The permanent input of a herbicidal compound into most vulnerable ecosystems might lead to adverse effects to biota (plants, microorganisms, etc.). TCA soil levels of coniferous forests in mountainous regions of Central Europe are significantly elevated. Mass balance calculations show that precipitation as sole source of TCA in soil seems to be of minor importance and provide evidence for a natural formation of TCA within soil itself. In addition, the isolation of a chlorinating enzyme in soil and laboratory experiments with humic acid, iron and halide point to an omnipresent chlorinating capability of nature producing polyhalogenated organic compounds such as TCA. In this paper we present an overview of TCA levels in the environment and provide a new estimate about the extent of a natural TCA formation, especially in soil. PMID- 12738258 TI - Review of concentrations and chemistry of trichloroacetate in the environment. AB - This paper reviews the concentrations of trichloroacetate (TCA) in the atmosphere plant-soil system. Data originate mainly from Europe. The median TCA concentration in rainwater and canopy drip decreased until 1995. From then the median TCA concentration in rainwater remains rather constant while for canopy drip later data are not available. The same seems to hold for concentrations in air although a very limited data set is available. The median concentrations in coniferous needles and groundwater are constant for the period observed. The median TCA concentrations in soil decreased until 1992 and then remained constant.The TCA formation from chlorinated solvents in the atmosphere may explain a substantial percentage of the TCA amount in the atmosphere. The TCA concentrations in rainwater and canopy drip indicate that there will be other sources contributing to 10-50%. Waste incineration, biomass burning and natural formation in the marine boundary layer are potential candidate sources of TCA, but nothing can be said as yet on their TCA emission rates. Anthropogenic emissions of chlorine could also be a source.TCA can be formed from chlorinated solvents by biota. However, for coniferous trees the uptake of TCA from soil may be the predominant route. Biotic and abiotic reactions can cause to formation of TCA in soil, but also formation of TCA from chlorinated solvents by biota that excrete TCA, may contribute. Mass balance calculations of the bioactive soil top layer show that the production rate of TCA in certain soil types could be substantial. The mass balance calculations could not distinguish between natural and anthropogenic sources in soil. PMID- 12738259 TI - Sense or no-sense of the sum parameter for water soluble "adsorbable organic halogens" (AOX) and "absorbed organic halogens" (AOX-S18) for the assessment of organohalogens in sludges and sediments. AB - "AOX" is the abbreviation of the sum parameter for water soluble "adsorbable organic halogens" in which 'A' stands for adsorbable, 'O' for organic and 'X' for the halogens chlorine, bromine and iodine. After the introduction of the AOX in 1976, this parameter has been correctly used for "real" AOX constituents (DDT and its metabolites, PCBs, etc.) but also misused for non-adsorbable adsorbed OX compounds, mostly high molecular organohalogens in plants and even to inorganic compounds being neither organic nor adsorbable. The question of natural "Adsorbable Organic Halogens" (AOX) formed by living organisms and/or during natural abiogenic processes has been definitively solved by the known existence of already more than 3650 organohalogen compounds, amongst them the highly reactive, cancerogenic vinyl chloride (VC). The extension of the AOX to AOX-S18 for Sludges and Sediments, in which A stands for adsorbed (not for adsorbable) is questionable. It includes the most important water insoluble technical organochlorine product: polyvinyl chloride, PVC. In addition to organic halogens it also includes inorganic, mineralogenic halides, incorporated mainly in the crystal lattice of fine grained phyllosilicates, the typical clay minerals (kaolinite, montmorillonite, illite and chlorite) which are main constituents of sediments and sedimentary rocks representing the major part of the sedimentary cover of the earth. Other phyllosilicates, biotite and muscovite, major constituents of granites and many metamorphic rocks (gneiss and mica schist) will also contribute to the AOX-S18 especially in soils as result of weathering processes. Since chlorine is incorporated into the mineral structure and, as a consequence, not soluble by the nitric acid analytical step (pH 0.5) of the S18 determination, it will account to the AOX-S18 in the final charcoal combustion step at temperatures >950 degrees C. After heavy rainfalls sewage sludge composition is strongly influenced by mineralogenic components derived from the erosion of fine grained sediments or soils. Assuming 50% geogenic particles with a mean Cl concentration of 103 mg/kg (as in shales and clays) the mineralogenic Cl-content could add about 50 mg/kg to the organic AOX in sewage sludge. The occurrence of insoluble and non-adsorbable PVC in sewage sludge exhibits the same problems as the mineralogenic constituents: a detection as AOX-S18 is possible when the final high temperature analytical step is applied. Plants as major sources of organohalogens have never been doubted. Only recently [Science 295 (2002) 985] based on the determination of the form of Cl with near-edge X-ray adsorption fine structure (NEXAFS) spectroscopy and extended X-ray adsorption showed the variations in the inorganic Cl(-) and organo-Cl compounds with increasing humification of plant leaves from "fresh leaves--senescent leaves on plants--senescent leaves on soil--powdered top soil--isolated soil humus". His finding of exclusively inorganic Cl(-) in the starting material (fresh leaves) is controverse to our earlier results indicating the presence of ionic inorganic Cl together with water insoluble absorbed organohalogens (AOX-S18) in eight different macrophytes of both terrestrial and marine environments. Our research on AOX in interstitial water of anaerobic limnic sediments has led to the role of bromine playing in the diagenesis of the organic matter of sediments. In sediments of Lake Constance Br(-) concentrations in lake water at the sediment water interface increased from <0.01 to 0.25 mg/l in the pore water at 77 cm sediment depth. In the Neckar River a Br concentrations of 0.02 mg/l at the water/sediment interface increasing to 0.74 mg/l in pore water in 85 cm depth was found. Here a parallel development could be found with ammonium concentration and alkalinity. The very high positive correlation ammonium:bromide and bromide:alkalinity leads to the conclusion, that bromine, originally a high molecular constituent of the organic matter, is released as bromide during an early dehalogenation stage of diagenesis. The mlusion, that bromine, originally a high molecular constituent of the organic matter, is released as bromide during an early dehalogenation stage of diagenesis. The main general reason to discard the AOX sum parameter as a whole lies in the fact, that adsorbable halogenated substances cannot a priori be categorized as natural/anthropogenic, biotic/abiotic, harmful/harmless. If applied to sludges and sediments, adsorbed organohalogens are not water soluble and therefore not adsorbable, and mineralogenic halogens (X) are neither organic nor adsorbable, and therefore by definition no AOX. PMID- 12738260 TI - Chloride imbalances in soil lysimeters. AB - The assumption that soil neither acts as a source or a sink of chloride is evaluated by incubating soil cores in lysimeters in a climate chamber under controlled conditions. Some of the lysimeters acted as a sink while others acted as a source of chloride. Considerable amounts of organic chlorine were lost by leaching. The loss by leaching of organic chlorine could only explain part of the discrepancy in the lysimeters where the soil acted as a sink and it could certainly not explain the cases where the soil acted as a source. The storage of organic chlorine was four times larger than the storage of chloride and comparably small changes in the organic chlorine storage will thus have a considerable influence on the chloride budget. However, the soil was too heterogeneous to determine whether a change in the storage had taken place or not. It is concluded that the observed chloride surplus and also, at least to some extent, the observed chloride deficit, most likely was caused by net-changes in the storage of organic chlorine in soil. An inverse correlation was found between the initial chloride content of the soil and the imbalance in the chloride budget.Dry deposition of chloride is generally assumed to equal the run off minus the wet deposition. Extrapolation to the field situation suggests that the output of organic chlorine by soil leachate is at risk to cause an underestimation of the dry deposition by about 25%. PMID- 12738261 TI - Spatial patterns of organic chlorine and chloride in Swedish forest soil. AB - The concentration of organic carbon, organic chlorine and chloride was determined in Swedish forest soil in the southern part of Sweden and the spatial distribution of the variables were studied. The concentration of organically bound chlorine was positively correlated to the organic carbon content, which is in line with previous studies. However, the spatial distribution patterns strongly indicate that some other variable adds structure to the spatial distribution of organic chlorine. The distribution patterns for chloride strongly resembled the distribution of organic chlorine. The spatial distribution of chloride in soil depends on the deposition pattern which in turn depends on prevailing wind-direction, amount of precipitation and the distance from the sea. This suggests that the occurrence of organic chlorine in soil is influenced by the deposition of chloride or some variable that co-varies with chloride. Two clearly confined strata were found in the area: the concentrations of organic chlorine and chloride in the western area were significantly higher than in the eastern area. No such difference among the two areas was seen regarding the carbon content. PMID- 12738262 TI - Occurrence and source of brominated organic compounds in surface waters. AB - Monitoring of organic halogen compounds, measured as adsorbable organically bound bromine (AOBr), in an eutrophic lake, which is influenced by treated waste water, revealed repeatedly high concentrations of organobromine compounds in late summer, whereas five times lower values were measured during the rest of the year. It was possible to reproduce the in situ observed AOBr increase in the laboratory. Batch experiments were performed with lake water from two different lakes and an algae culture. It could be shown that the AOBr production is not limited to strong waste water influenced lakes. Furthermore, the AOBr formation requires light and the presence of algae, and thus is most probably biotic in nature. A low content of nutrients favours the formation of organic bromine compounds. To our knowledge this is the first report about the seasonally occurrence of naturally produced organic bromine compounds in lakes/surface waters. PMID- 12738263 TI - Di- and tribromoindoles in the common oyster (Crassostrea virginica). AB - The sources/origins, fate and impacts of naturally occurring organobromine compounds in the marine environment are largely unknown. Soft tissue composites of the common oyster (Crassostrea virginica) collected from coastal Georgia (USA) were analyzed for organobromines by gas chromatography. Three simple bromoindoles (BIs)--two dibromo- and one tribromo congener--were detected and their molecular formulas elucidated by electron and negative chemical ionization GC-MS. Semi quantitative estimates of BI concentrations in these samples using GC-ECD indicated that oysters sampled in November 1999 contained 5-10 times more BIs than those sampled in August 2000 and March 2001. Although their bromine substitution patterns are presently unknown, this first ever report of bromoindoles in C. virginica, a prolific and important commercial and ecological species, underscores its potential utility as a bioindicator of organobromines in the coastal marine environment. PMID- 12738264 TI - Non-polar halogenated natural products bioaccumulated in marine samples. I. 2,3,3',4,4',5,5'-Heptachloro-1'-methyl-1,2'-bipyrrole (Q1). AB - This presentation adds new spectroscopic and analytical data on the natural product Q1 that was recently identified by synthesis as 2,3,3('),4,4('),5,5(') heptachloro-1(')-methyl-1,2(')-bipyrrole. Solid state magic angle spinning 13C NMR data of Q1 is presented as an option for structural proof. Furthermore, the UV spectrum of neat Q1 (absorption maximum at 223 nm) was recorded and, with NMR spectroscopic data, confirmed a twisted bipyrrole ring system. A quantitative standard of Q1 was prepared which allowed to correct previous concentration estimates relative to the electron capture detector response factor of trans nonachlor. As a result, the actual Q1 response was only 0.65+/-15% of the response factor of trans-nonachlor. Therefore, actual Q1 levels are about 50% higher than the previous estimates. With this result the highest (corrected) Q1 concentration determined to date in the blubber of marine mammals from Australia is 14 mg/kg lipid. Analysis of Q1 and trans-nonachlor in specimens from the German North Sea coast suggests that harbor seals are more able to metabolize Q1 than harbor porpoises. Finally, we calculated that 79 congeners of Q1 (i.e. lower chlorinated 1(')-methyl-1,2(')-bipyrroles) are theoretically possible and present their structures. PMID- 12738265 TI - Non-polar halogenated natural products bioaccumulated in marine samples. II. Brominated and mixed halogenated compounds. AB - Several identified and potential natural brominated bioaccumulative compounds were studied in this work. 4,6-dibromo-2-(2('),4(')-dibromo)phenoxyanisole (BC-2) previously detected in Australian marine mammals and isolated from sponges, was synthesized. Two byproducts (a tetrabromo isomer and a tribromo congener) were investigated as well. The byproducts of the synthesis were not identified in the environmental samples investigated. Previously described natural brominated compounds (BC-1, BC-2, BC-3, BC-10, BC-11, MHC-1) and anthropogenic brominated diphenyl ethers (BDE-47, BDE-99, BDE-100, BDE-154) were detected in a sample of human milk. The sample was from a woman from the Faeroe Islands who frequently consumed fish as well as whale blubber and meat. The most abundant compound originated from the natural tetrabromo phenoxyanisole BC-3 which may have a 3:1 distribution of bromine on the two phenyl units. This sample also accumulated a dibromochloroanisole, as well as a previously unknown mixed halogenated compound (MHC-X) and an unknown, most likely aromatic brominated compound. Co-elutions on a DB-5 column were found for BDE-99 and BC-11 as well as BDE-154 and the unknown brominated compound. This suggests that quantification of these two compounds has to be carried out carefully.Two samples of lower trophic level, namely Baltic cod liver and Mexican mussel tissue, were investigated as well. The cod liver samples contained BDE congeners but also abundant signals for the natural 2,3,3('),4,4('),5,5(')-heptachloro-1(')-methyl-1,2(')-bipyrrole Q1 and tribromoanisole (TBA). The mussel sample contained Q1, TBA, another halogenated anisole, BC-1, BC-2, and BC-3, as well as additional, potential natural brominated compounds in the elution range of tribromophenoxyanisoles. PMID- 12738266 TI - The distinctive isotopic signature of plant-derived chloromethane: possible application in constraining the atmospheric chloromethane budget. AB - Chloromethane (CH(3)Cl) is the most abundant halocarbon in the atmosphere. Although largely of natural origin it is responsible for around 17% of chlorine catalysed ozone destruction. Sources identified to date include biomass burning, oceanic emissions, wood-rotting fungi, higher plants and most recently tropical ferns. Current estimates reveal a shortfall of around 2 million ty(-1) in sources versus sinks for the halocarbon. It is possible that emissions from green plants have been substantially underestimated. A potentially valuable tool for validating emission flux estimates is comparison of the delta13C value of atmospheric CH(3)Cl with those of CH(3)Cl from the various sources. Here we report delta13C values for CH(3)Cl released by two species of tropical ferns and show that the isotopic signature of CH(3)Cl from pteridophytes like that of CH(3)Cl from higher plants is quite different from that of CH(3)Cl produced by biomass burning, fungi and industry. delta13C values for CH(3)Cl produced by Cyathea smithii and Angiopteris evecta were respectively -72.7 per thousand and 69.3 per thousand representing depletions relative to plant biomass of 42.3 per thousand and 43.4 per thousand. The characteristic isotopic signature of CH(3)Cl released by green plants should help constrain their contribution to the atmospheric burden when reliable delta13C values for all other major sources of CH(3)Cl are obtained and a globally averaged delta13C value for atmospheric CH(3)Cl is available. PMID- 12738267 TI - Uptake, translocation and fate of trichloroacetic acid in a Norway spruce/soil system. AB - Trichloroacetic acid (TCA) is a secondary atmospheric pollutant formed by photooxidation of chlorinated solvents in the troposphere--it has, however, recently been ranked among natural organohalogens. Its herbicidal properties might be one of the factors adversely affecting forest health. TCA accumulates rapidly in conifer needles and influences the detoxification capacity in the trees. The aim of the investigations--a survey of which is briefly given here- was to elucidate the uptake, distribution and fate of TCA in Norway spruce. For this purpose young nursery-grown plants of Norway spruce (Picea abies (L.) Karst.) were exposed to [1,2-14C]TCA and the fate of the compound was followed in needles, wood, roots, soil and air with appropriate radio-indicator methods. As shown by radioactivity monitoring, the uptake of TCA from soil by roots proceeded most rapidly into current needles at the beginning of the TCA treatment and was redistributed at later dates so that TCA content in older needles increased. The only product of TCA metabolism/biodegradation found in the plant/soil-system was CO(2) (and corresponding assimilates). TCA biodegradation in soil depends on TCA concentration, soil humidity and other factors. PMID- 12738269 TI - Peatlands: a major sink of naturally formed organic chlorine. AB - It is a little known fact that many chlorinated organic compounds occur naturally and that some are also indispensable to life on earth. Here, we show that chlorination of organic compounds during humification processes in peat is widespread in nature. Globally this process has led to the accumulation of approximately 280-1000 million tons of organically bound chlorine in peatlands during the postglacial period. PMID- 12738268 TI - Input of trichloroacetic acid into the vegetation of various climate zones- measurements on several continents. AB - Trichloroacetic acid (TCA, CCl(3)COOH) is a phytotoxic chemical. Although TCA salts and derivates were once used as herbicides to combat perennial grasses and weeds, they have since been banned because of their indiscriminate herbicidal effects on woody plant species. However, TCA can also be formed in the atmosphere. For instance, the high-volatile C(2)-chlorohydrocarbons tetrachloroethene (TECE, C(2)Cl(4)) and 1,1,1-trichloroethane (TCE, CCl(3)CH(3)) can react under oxidative conditions in the atmosphere to form TCA and other substances. The ongoing industrialisation of Southeast Asia, South Africa and South America means that use of TECE as solvents in the metal and textile industries of these regions in the southern hemisphere can be expected to rise. The increasing emissions of this substance--together with the rise in the atmospheric oxidation potential caused by urban activities, slash and burn agriculture and forest fires in the southern hemisphere--could lead to a greater input/formation of TCA in the vegetation located in the lee of these emission sources. By means of biomonitoring studies, the input/formation of TCA in vegetation was detected at various locations in South America, North America, Africa, and Europe. PMID- 12738270 TI - Fluorinated natural products: the biosynthesis of fluoroacetate and 4 fluorothreonine in Streptomyces cattleya. AB - Organofluorine compounds are rare in Nature, with only a handful known to be produced by some species of plant and two microorganisms. Consequently, the mechanism of enzymatic carbon-fluorine bond formation is poorly understood. The bacterium Streptomyces cattleya biosynthesises fluoroacetate and 4 fluorothreonine as secondary metabolites and is a convenient system to study the biosynthesis and enzymology of fluorometabolite production. Using stable-isotope labelled precursors it has been shown that there is a common intermediate in the biosynthesis of the fluorometabolites, which has recently been identified as fluoroacetaldehyde. Studies with cell-free extracts of S. cattleya have identified two enzymes, an aldehyde dehydrogenase and a threonine transaldolase, that are involved in the biotransformation of fluoroacetaldehyde to fluoroacetate and 4-fluorothreonine. PMID- 12738271 TI - UV-light induced mineralization of organic matter bound chlorine in Lake Bjan, Sweden--a laboratory study. AB - Surface water and aqueous solutions of isolated organic matter from a humic rich lake in southern Sweden were exposed to artificial UV radiation to investigate the UV light induced influence on organic matter bound chlorine in natural systems. It was found that the photodegradation of organic matter bound chlorine was more pronounced than the photodegradation of organic carbon. After 120 h of irradiation of the isolated organic matter, only 35% of the initial organochlorine was still in the solution compared to about 70% of the dissolved organic carbon (DOC). A similar result was obtained for unfractionated surface water. Furthermore, our results indicate that the loss of organic chlorine was mainly due to a mineralization of organic chlorine into chloride ions. The total decrease of organic chlorine after 120 h was 32 microg Cl(org) l(-1), of which the major part disappeared in the initial irradiation phase. A similar increase was observed in the chloride concentration (34 microg Cl(-) l(-1)). PMID- 12738272 TI - Do vesicle cells of the red alga Asparagopsis (Falkenbergia stage) play a role in bromocarbon production? AB - The Rhodophyceae (red algae) are an established source of volatile halocarbons in the marine environment. Some species in the Bonnemaisoniaceae have been reported to contain large amounts of halogens in structures referred to as vesicle cells, suggesting involvement of these specialised cells in the production of halocarbons. We have investigated the role of vesicle cells in the accumulation and metabolism of bromide in an isolate of the red macroalga Asparagopsis (Falkenbergia stage), a species known to release bromocarbons. Studies of laboratory-cultivated alga, using light microscopy, revealed a requirement of bromide for both the maintenance and formation of vesicle cells. Incubation of the alga in culture media with bromide concentrations below 64 mgl(-1) (the concentration of Br(-) in seawater) resulted in a decrease in the proportion of vesicle cells to pericentral cells. The abundance of vesicle cells was correlated with bromide concentration below this level. Induction of vesicle cell formation in cultures of Falkenbergia occurred at concentrations as low as 8 mgl(-1), with the abundance of vesicle cells increasing with bromide concentration up to around 100 mgl(-1). Further studies revealed a positive correlation between the abundance of vesicle cells and dibromomethane and bromoform production. Interestingly, however, whilst dibromomethane production was stimulated by the presence of bromide in the culture media, bromoform release remained unaffected suggesting that the two compounds are formed by different mechanisms. PMID- 12738273 TI - Formation of volatile iodinated alkanes in soil: results from laboratory studies. AB - Volatile iodinated organic compounds play an important role in the tropospheric photochemical system, but the current knowledge of the known sources and sinks of these alkyl iodides is still incomplete. This paper describes a new source of alkyl iodides from the pedosphere. Different organic-rich soils and humic acid were investigated for their release of volatile organoiodides. Six volatile organoiodides, iodomethane, iodoethane, 1-iodopropane, 2-iodopropane, 1 iodobutane and 2-iodobutane were identified and their release rates were determined. We assume an abiotic reaction mechanism induced by the oxidation of organic matter by iron(III). The influence of iron(III), iodide and pH on the formation of alkyl iodides was investigated. Additionally, different organic substances regarded as monomeric constituents of humus were examined for the production of alkyl iodides. Two possible reaction pathways for the chemical formation of alkyl iodides are discussed. As humic acids and iron(III) are widespread in the terrestrial environment, and the concentration of iodide in soil is strongly enriched (compared to seawater), this soil source of naturally occurring organoiodides is suggested to contribute significantly to the input of iodine into the troposphere. PMID- 12738274 TI - Halogen production from aqueous tropospheric particles. AB - Box model studies have been performed to study the role of aqueous phase chemistry with regard to halogen activation for marine and urban clouds and the marine aerosol as well. Different chemical pathways leading to halogen activation in diluted cloud droplets and highly concentrated sea salt aerosol particles are investigated. The concentration of halides in cloud droplets is significantly smaller than in sea-salt particles, and hence different reaction sequences control the overall chemical conversions. In diluted droplets radical chemistry involving OH, NO(3), Cl/Cl(2)(-)/ClOH(-), and Br/Br(2)(-)/BrOH(-) gains in importance and pH independent pathways lead to the release of halogens from the particle phase whereas the chemistry in aerosol particles with high electrolyte concentrations is controlled by non-radical reactions at high ionic strengths and relatively low pH values. For the simulation of halogen activation in tropospheric clouds and aqueous aerosol particles in different environments a halogen module was developed including both gas and aqueous phase processes of halogen containing species. This module is coupled to a base mechanism consisting of RACM (Regional Atmospheric Chemistry Mechanism) and the Chemical Aqueous Phase Radical Mechanism CAPRAM 2.4 (MODAC-mechanism). Phase exchange is described by the resistance model by Chemistry of Multiphase Atmospheric Systems, NATO ASI Series, 1986. It can be shown that under cloud conditions the bromine atom is mainly produced by OH initiated reactions, i.e. its concentration maximum is reached at noon. In contrast, the concentration level of chlorine atoms is linked to NO(3) radical chemistry leading to a smaller amplitude between day and night time concentrations. The contribution of radical processes to halogen atom formation in the particle phase is evident, e.g. by halogen atoms which undergo direct phase transfer. Furthermore, the application of the multiphase model for initial concentrations for sea-salt aerosols shows that the particle phase can act as a main source of halogen containing molecules (Cl(2), BrCl, Br(2)) which are photolysed in the gas phase to yield halogen atoms (about 70% of all Cl sources and more than 99% for Br). PMID- 12738275 TI - Stereoselective biosynthesis of chloroarylpropane diols by the basidiomycete Bjerkandera adusta. AB - Previously we have shown that 1-arylpropane-1,2-diols are catabolic products of L phenylalanine during idiophasic metabolism of B. adusta that are stereoselectively biosynthesized from a C(7)-unit (ring+benzylic carbon) and a C(2)-unit as predominantly erythro 1R, 2S enantiomers.In order to probe the mechanism of 1-arylpropane-1,2-diol formation, the products of the incubation of isotopically labelled aromatic aldehydes as substrates with Bjerkandera adusta (DAOM 215869) have been characterized. The aromatic aldehydes were benzaldehyde (ring D(5)) and 4-methoxy- and 4-hydroxybenzaldehydes (ring 13C(6)). These aldehydes were all stereoselectively incorporated into the corresponding 1 arylpropane-1,2-diols, including the chloro analogues, as well as into the corresponding alpha-ketols (phenyl acetyl carbinols (PAC's) and 2-hydroxy propiophenones (2-HPP's)) the presumed precursors of the diols. Benzoic acid (ring D(5)) was likewise incorporated into the diols, chlorodiols and alpha ketols. These results lead us to conclude that the aromatic aldehydes benzaldehyde, 4-hydroxybenzaldehyde and 4-methoxybenzaldehyde are likely C(7) unit precursors in the carboligation reaction(s) that leads to 1-arylpropane-1,2 diol biosynthesis. The metabolic role of the diols remains to be elucidated but they may be important intermediates in CAM (chlorinated anisyl metabolite) aldehyde-alcohol cycling and also act as substrates for the chlorination/hydroxylation enzymes yet to be identified in white rot fungi. PMID- 12738277 TI - Scientific activities of Euro Chlor in monitoring and assessing naturally and man made organohalogens. AB - In this paper a review of the scientific activities and research programmes carried out by Euro Chlor, the European Federation of chlor-alkali producers is presented according to two main axes: marine risk assessments with statistical analysis of monitoring data, temporal trends of emission levels and environmental concentrations. The methodology applied in each field is briefly presented and then illustrated by several practical examples. As a large part of the uncertainties in assessing the risk of a chemical to a given species or ecosystem often comes from the difficulty in evaluating the exposure level, Euro Chlor has chosen to use a monitoring approach, the exposure level being estimated from a statistical analysis of measured concentrations levels in water and sediment from rivers, estuaries and coastal areas. As the modelling approach often used by the authorities to estimate the predicted environmental concentration value is starting from roughly estimated emission levels, Euro Chlor collated emissions data from about 80 production plants in order to reduce the uncertainty associated with the default values introduced in the modelling approach.A brief review of the European emission levels for chlorinated organic substances is given as well as the temporal trends of both emission and environmental levels. A methodology to quantify the trends in measured concentrations at local and regional scales is briefly described. The observed decreasing trends demonstrate the continuous progress made by the Euro Chlor member companies in protecting the environment.Finally, the problems linked to the simultaneous presence in the environment of naturally and man-made chlorinated substances are briefly reviewed. To stimulate further research in the field, two key questions are raised which have not yet found a satisfactory answer: how to quantify natural background levels and how to quantify global persistence in the environment? PMID- 12738276 TI - Formation of chloroacetic acids from soil, humic acid and phenolic moieties. AB - The mechanism of formation of chloroacetates, which are important toxic environmental substances, has been controversial. Whereas the anthropogenic production has been well established, a natural formation has also been suggested. In this study the natural formation of chloroacetic acids from soil, as well as from humic material which is present in soil and from phenolic model substances has been investigated. It is shown that chloroacetates are formed from humic material with a linear relationship between the amount of humic acid used and chloroacetates found. More dichloroacetate (DCA) than trichloroacetate (TCA) is produced. The addition of Fe(2+), Fe(3+) and H(2)O(2) leads to an increased yield. NaCl was added as a source of chloride. We further examined the relationship between the structure and reactivity of phenolic substances, which can be considered as monomeric units of humic acids. Ethoxyphenol with built-in ethyl groups forms large amounts of DCA and TCA. The experiments with phenoxyacetic acid yielded large amounts of monochloroacetate (MCA). With other phenolic substances a ring cleavage was observed. Our investigations indicate that chloroacetates are formed abiotically from humic material and soils in addition to their known biotic mode of formation. PMID- 12738278 TI - Environmental risk assessment of airborne trichloroacetic acid--a contribution to the discussion on the significance of anthropogenic and natural sources. AB - In environmental risk assessments the question has to be answered, whether risk reduction measures are necessary in order to protect the environment. If the combination of natural and anthropogenic sources of a chemical substance leads to an unacceptable risk, the man-made emissions have to be reduced. In this case the proportions of the anthropogenic and natural emissions have to be quantified. Difficulties and possible solutions are discussed in the scope of the OECD- and EU-risk assessments of trichloroacetic acid (TCA) and tetrachloroethylene. In the atmosphere, TCA is formed by photo-oxidative degradation of tetrachloroethylene (PER) and 1,1,1-trichloroethane. The available data on atmospheric chemistry indicate that tetrachloroethylene is the more important pre-cursor. With its high water solubility and low volatility, TCA is adsorbed onto aerosol particles and precipitated during rainfalls. Extended monitoring in rainwater confirmed the global distribution of airborne TCA. TCA reaches soils by dry and wet deposition. In addition formation of TCA from tetrachloroethylene in plants was observed. Consequently, high concentrations were detected in needles, leaves and in forest soil especially in mountain regions. The effect assessment revealed that plants exposed via soil are the most sensitive species compared to other terrestrial organisms. A PNECsoil of 2.4 microg/kg dw was derived from a long-term study with pine and spruce seedlings. When this PNEC is compared with the measured concentrations of TCA in soil, in certain regions a PEC/PNEC ratio >1 is obtained. This clearly indicates a risk to the terrestrial ecosystem, with the consequence that risk reduction measures are deemed necessary. To quantify the causes of the high levels of TCA in certain soils, and to investigate the geographical extent of the problem, intensive and widespread monitoring of soil, air and rainwater for TCA and tetrachloroethylene would be necessary to be able to perform a full mass balance study at an appropriate number of sites. In addition, measurements of the 14C content in TCA isolated from soil could clarify whether a significant proportion of the TCA occurs from natural sources. The possible formation of TCA in soil can also be tested by incubation of isotope enriched inorganic chloride with subsequent mass spectrometry of TCA. PMID- 12738279 TI - Functional activation of arylhydrocarbon receptor (AhR) in primary T cells by 2,3,7,8-tetrachlorodibenzo-p-dioxin. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) exerts diverse adverse health effects by activating the transcription factor arylhydrocarbon receptor (AhR). The activated AhR induces the expression of various genes having xenobiotic responsive elements (XREs) in their enhancer regions, such as the gene for cytochrome P-450 1A1 (CYP1A1). The immune system is sensitively affected by TCDD, while the precise mechanism of how TCDD acts in each immune cell type remains to be determined. The results of previous studies on AhR activation in B cell lines, T cell clones, and thymocytes, which mainly consist of immature T cells, suggested that AhR in mature T cells is inactive, whereas that in B cells and immature T cells act functionally. In the present study, we investigated whether or not TCDD induces the CYP1A1 gene by functionally activating AhR in primary mature T cells in mice. When the splenocytes that contain mature T and B lymphocytes as their predominant cell types or the thymocytes were cultured in the presence of TCDD, each of them showed a similar magnitude of CYP1A1 induction with a peak induction at 4 h. Both mature T cells and B cells that had been separated from total splenocytes also showed CYP1A1 induction at the same magnitude with a peak induction at 4 h. Gene expression of CYP1A1 was observed at 0.1 nM or greater concentrations of TCDD in splenocytes and separated T cells. The induction of CYP1A1 in T cells was confirmed in mice exposed to TCDD. These results indicate that TCDD induces the functional activation of AhR in primary mature T cells in mice. PMID- 12738280 TI - Biodegradation capability of Absidia fusca Linnemann towards environmental pollutants. AB - The purpose of this work was to study the bioremediation capability of Absidia fusca Linnemann (Zygomycete) towards different classes of xenobiotics (lignin derived compounds, chloroaromatic compounds, polycyclic aromatic hydrocarbons) the presence of which in contaminated soils, water and sediments poses a significant risk to the environment and human health. Two strains from different origins were compared. One was from an official collection and grown in non inducing conditions, while the other was isolated during the course of the survey of fungal flora in a polluted soil from Annaba (Algeria). All data were analyzed and results validated via a statistical treatment. We showed the effect of the factors studied (origin of the strain, xenobiotic) but also the interactions between these factors. The strain of A. fusca isolated from a polluted soil was able to efficiently degrade most of the xenobiotics tested, particularly: pentachlorophenol, phenol, catechol, guaiacol and ferulic acid. This property also existed in the other strain but at a very low level. PMID- 12738281 TI - Biodegradation of phthalate esters during the mesophilic anaerobic digestion of sludge. AB - Phthalic acid esters (PAE) are commonly found in the sludge generated in the wastewater treatment plants. Anaerobic digestion followed by land application is a common treatment and disposal practice of sludge. To date, many studies exist on the anaerobic biodegradation rates of PAE, especially of the easily biodegradable ones, whereas the higher molecular weight PAE have reported to be non-biodegradable under methanogenic conditions. Furthermore, there is no information on the effect of the PAE on the performance of the anaerobic digesters treating sludge. In this study, the anaerobic biodegradation of di-n butyl phthalate (DBP), di-ethyl phthalate (DEP) and di-ethylhexyl phthalate (DEHP) was investigated and their relative rates of anaerobic degradation were calculated. Also, the biological removal of PAE during the anaerobic digestion of sludge in bench-scale digesters was investigated using DBP and DEHP as model compounds of one biodegradable and one recalcitrant PAE respectively. The degradation of all the PAE tested in this study (DEP, DBP and DEHP) is adequately described by first-order kinetics. Batch and continuous experiments showed that DEP and DBP present in sludge are rapidly degraded under mesophilic anaerobic conditions (a first-order kinetic constant of 8.04 x 10(-2) and 13.69 x 10(-2) 4.35 day(-1) respectively) while DEHP is degraded at a rate between one to two orders of magnitude lower (0.35 x 10(-2)-3.59 x 10(-2) day(-1)). It is of high significance that experiments with anaerobic sludge of different origin (US and Europe) showed that degradation of DEHP occurs under methanogenic conditions. Accumulation of high levels of DEHP (more than 60 mg/l) in the anaerobic digester has a negative effect on DBP and DEHP removal rates as well as on the biogas production. PMID- 12738283 TI - Development of expanded and core kinetic models for the gas phase formation of dioxins from chlorinated phenols. AB - Expanded, 45 reaction, and core, 12 reaction, kinetic models have been developed that account for the major features in the homogeneous formation of polychlorinated dibenzo-p-dioxins (PCDD) from the oxidation of 2,4,6 trichlorophenol (P). The expanded and core schemes provide good agreement between experimental and calculated yields of PCDDs using the CHEMKIN combustion package or the React kinetic program, respectively. Steady-state approximations of the reaction kinetic models including radical-molecule and radical-radical formation pathways of PCDD, as well as oxidative destruction pathways of chlorinated phenoxyl radicals, reveal a competition between reactions of chlorinated phenoxyl radicals with chlorinated phenols, recombination reactions of chlorinated phenoxyl mesomers, and destruction/decomposition of phenoxyl radicals. PMID- 12738282 TI - Seasonal change of persistent organic pollutant concentrations in air at Niigata area, Japan. AB - The concentrations of persistent organic pollutants (POPs), such as HCB, alpha-, beta-, gamma- and delta-HCH, trans- and cis-chlordane (t-CHL, c-CHL), DDE, DDD and DDT, in ambient air have been measured at five sampling points in Niigata area, Japan (Niigata, Maki, Tsubame, Jouzo and Yahiko) during the period from September 1999 to November 2001. HCB, alpha-HCH, t-CHL and c-CHL showed higher concentrations than the other chemicals in all locations. All the POPs except t CHL and c-CHL collected at urban sites of the Niigata Plain was almost the same in their concentration levels. Higher concentrations of t-CHL and c-CHL in residential areas should be attributed to the past usage of the chemical as a termiticide. At Yahiko (remote site), most of the POPs showed lower concentrations than those measured at the other sampling sites, although alpha HCH and gamma-HCH were comparable with the concentrations found at the other sampling sites. All POPs except alpha-HCH and gamma-HCH tend to decrease 41-80% in their concentrations from 2000 to 2001. The lower POPs concentrations in winter and the higher POPs concentrations in summer at every sampling point can be partly explained by temperature differences. Applying the equation of the logarithm of the POP partial pressure in air versus reciprocal temperature (lnPa=m/T+b) to our data, linear relations were observed. HCB gave a poor linearity and the smallest slope, while beta-HCH, t-CHL and c-CHL gave good linearities and large slopes in the equation. The results suggest that HCB level is influenced by not only the emission from terrestrial sources but the global scale background pollution. A peculiar observation is that beta-HCH concentration measured in our study showed large temperature dependence, indicating there could be a source of contamination in the surrounding areas. PMID- 12738284 TI - Formation of chlorinated hydrocarbons from the reaction of chlorine atoms and activated carbon. AB - The reactions of chlorine atoms and activated carbon have been studied over the temperature range of 200-400 degrees C using an isothermal flow reactor in conjunction with 337 nm laser photolysis of Cl2. These studies have shown that carbon tetrachloride is the major product, with chloroform, methylene chloride, and methyl chloride being formed in progressively decreasing yields. Trace quantities of methane, ethane, and dichloroethylenes were also observed. Mechanisms of carbon fragmentation by successive addition of chlorine atoms are proposed. The formation of small chlorinated hydrocarbons by the direct reaction of chlorine with carbon may be a key link in both the de novo and precursor pathways of formation of PCDD/F. PMID- 12738285 TI - DDT is still a problem in developed countries: the heavy pollution of Lake Maggiore. AB - The Zebra mussel (Dreissena polymorpha), one of the most widely used bioindicators of persistent organic pollutants, trace metals and radionuclides in several worldwide freshwater ecosystems, has been used to monitor DDT contamination trends in Lake Maggiore since 1996, caused by industrial effluents on a tributary of the River Toce, one of the major affluents of the lake. Dreissena specimens were collected at two sampling sites, one within the Baveno Bay, where the River Toce flows, and the other outside (Villa Taranto). Total DDT levels (3119.6 ng/g lipids at Baveno and 1351.2 ng/g lipids at Villa Taranto) in the soft tissues of the Zebra mussel decreased at both stations by about 30-50% in the first year after the closure of the chemical plant reaching an almost steady-state condition. The high concentrations measured in Zebra mussel specimens of Baveno Bay in 2000 (1947 ng/g lipids) and the percentage of pp'DDE in comparison with total DDT concentration, which showed a slight increase in the last years, clearly indicate that a contamination source is still present, deriving probably from the lacustrine sediments and the River Toce. Data show that the environmental risk is very high within the Baveno Bay and the recovery time should be longer than in the other parts of the lake, where DDT levels in Dreissena are presently two times higher than those measured in the other Italian subalpine lakes. PMID- 12738286 TI - Survey of commercial pasteurised cows' milk in Wallonia (Belgium) for the occurrence of polychlorinated dibenzo-p-dioxins, dibenzofurans and coplanar polychlorinated biphenyls. AB - Congener-specific analyses of 7 polychlorinated dibenzo-p-dioxins (PCDDs), 10 polychlorinated dibenzofurans (PCDFs) and 4 non-ortho (coplanar) polychlorinated biphenyls (cPCBs) were performed on 35 samples of commercial long-life pasteurised cows' milk issued from eight different brands available in Walloon supermarkets (Belgium). The observed congener profile was characteristic of milk samples issued from industrialised countries with good inter and intra-brand reproducibility's. The PCDDs to PCDFs ratio was equal to 1.8 in concentration. The toxic equivalent (TEQ based on WHO-TEF) value for PCDD/Fs in all analysed milks was 1.09+/-0.30 pg TEQ/g fat (range 0.86-1.59), which is below the recommended EU non-commercialisation threshold value of 3 pg TEQ PCDD/Fs/g of milk fat. The mean TEQ value including cPCBs was 2.23+/-0.55 pg TEQ/g fat. These PCBs actually contributed for 49+/-8.6% of the total TEQ. Among PCDD/Fs and cPCBs, tetrachloro dibenzo-p-dioxin (TCDD), pentachloro dibenzo-p-dioxin (PeCDD), pentachloro dibenzofurans (PeCDFs) and 3,3',4,4',5-pentachloro biphenyl (PCB-126) were the most important contributors to the TEQ. Estimated daily intake (EDI) due to consumption of such milks was 0.34 pg TEQ/kg of body weight/day for PCDD/Fs and 0.69 pg TEQ/kg of body weight/day when cPCBs were included. PMID- 12738287 TI - Chlorobenzenes removal from municipal solid waste incineration fly ash by surfactant-assisted column flotation. AB - The organic contaminants in municipal solid waste incineration (MSWI) fly ash, including chlorinated aromatic compounds and polycyclic aromatic hydrocarbons, have high toxicity and a potential negative impact on the environment. An effective and low energy consumption technique to remove the organic contaminants from MSWI fly ash is required urgently. Organic contaminants, such as chlorobenzenes (CBzs), in MSWI fly ash are known to become enriched in the unburnt carbon (UC) fraction. It is proposed that removal of UC from fly ash will result in the effective removal of most organic micropollutants. In this research, we use a technique of surfactant-assisted column flotation to decontaminate MSWI fly ash by removal of the CBzs-enriched UC from MSWI fly ash. We find that 39.8% of CBzs can be removed from fresh MSWI fly ash with 61.7% UC removal efficiency, whereas only 33.2% of CBzs can be removed from weathered MSWI fly ash with a low UC removal efficiency of 33.7%. By adding a mixture of two kinds of surfactants: sorbitan mono-oleate and polyoxyethylene (20) sorbitan mono oleate to the weathered fly ash, 47.0% of CBzs were removed at the hydrophile lipophile balance value of 13.5, while the UC removal efficiency increased to 49.0%. The results show that surfactants can enhance CBzs and UC removal efficiencies during the column flotation process. Higher CBzs and UC removal efficiencies can be expected by further optimizing the conditions of surfactant assisted column flotation. PMID- 12738288 TI - Polychlorinated dibenzofurans/dibenzo-p-dioxins (PCDF/PCDDs) and other dioxin like substances in marine organisms from the Grenland fjords, S. Norway, 1975 2001: present contamination levels, trends and species specific accumulation of PCDF/PCDD congeners. AB - Discharge of WHO toxicity equivalents (TEQs) of PCDF/PCDDs to Frierfjorden, Norway has been reduced from an estimated sum of 50-100 kg in the period 1951 1975 to about 6-7 kg in 1976-1990, and further to about 20 g for 1991-2000. In accordance with this, the yearly monitoring since 1987 has shown considerably decreasing contamination in organisms, first highly significant in all indicator species from 1990 to 1991, then levelling off. Present concentrations thus are still high. Compared with estimated "high background" (reference) concentrations of 10 ng TEQ(PCDF/PCDD)/kg w.w. in liver of cod and in hepatopancreas ("brown meat") of crabs, Frierfjord samples in 2001 were about 60 and 70 times higher, respectively. With considerable uncertainty due to large fluctuations, the rate of yearly decrease for TEQ(PCDF/PCDD) in cod liver 1991-2001 has been calculated to 10-12%. A hypothetic target value of 50 ng TEQ/kg w.w. will not be reached until 2015-2020, possibly even later. Including contributions from dioxin-like PCBs and PCNs, the weekly maximum tolerable amount of cod liver and crab hepatopancreas from Frierfjorden in 2001 were about 2-3 g. Multivariate analysis of PCDF/PCDD congener profiles in four fish species, mussels and crabs resulted in five distinct groups, separating four of the species and grouping the remaining two together, hence demonstrating examples of species specific accumulation characteristics. PMID- 12738289 TI - Analyses of known and new types of polyhalogenated aromatic substances in oven ash from recycled aluminium production. AB - Persistent aromatic bromine, chlorine and mixed chlorine-bromine compounds were analysed from recycled aluminium smelter (ALS) ashes to explore the impact of brominated flame retardants (BFR) on their formation. Polybrominated diphenyl ethers (PBDE) were the most abundant original BFRs found. Induction furnace ash contained tetra- to octa-BDEs about 2000ng g(-1) in similar congener ratios as the original scrap, but contents of nona- and deca-BDEs were only 25 and 5ng g( 1) indicating their significant degradation in ALS process. In the most non-polar fraction, PCB levels and profiles were similar as earlier ALS ash samples in 1990s. The highest PCB level measured was that of deca-CB (450ng g(-1)) in the induction furnace ash. In this fraction, bromo compounds were non-detectable (<5ng g(-1)). Fraction of the most polar compounds (from reversed toluene elution of carbon column ("dioxin fraction") contained PCDDs, PCDFs and polychlorinated dibenzothiophenes (PCDTs) in similar amounts and congener profiles as earlier investigated ALS ash samples. Bromine-containing dioxin and furan congeners were not detected. From individual PCDDs and PCDFs, octaCDF was the most abundant (205ng g(-1)) in induction furnace ash. In this fraction, the original BFR, tetrabromo-bisphenol-A, was identified. Its level in the induction furnace ash was approximated to be 388ng g(-1). In addition, 12 novel brominated and chlorinated compounds were found as abundant (8-441ng g(-1) in the induction furnace ash) contaminants from the fraction. Four of them were bisphenol derivatives, five biphenylols, then octachlorofluorenone (OCFL) and octachlorobiphenylene (OCBP). Their structures or structure types were deduced from total low-resolution EI mass spectra by theoretical isotope cluster simulation (ICLU) and through known fragmentation rules. PMID- 12738290 TI - PCB congener distributions in muscle, liver and gonad of Fundulus heteroclitus from the lower Hudson River Estuary and Newark Bay. AB - Gradients in sediment polychlorinated biphenyl (PCB) concentrations and PCB congener profiles exist along the Hudson River (NY, USA). We evaluated site and tissue differences in PCB concentration and congener profiles in resident mummichog (Fundulus heteroclitus) collected from PCB-contaminated sites in the lower Hudson River and the New York/New Jersey Harbor Estuary. Fish were collected from three PCB-contaminated sites Piermont Marsh (P), Iona Marsh (I), and Newark Bay (NB), and from two reference sites (Flax Pond, NY; Succotash Salt Marsh, RI). Congener profiles were statistically analyzed using principal component analysis (PCA) and general linear model (GLM) profile analysis. Contaminated fish had PCB tissue concentrations approximately 10-fold higher than those of reference fish. There were no site differences in PCB body burden (all tissues combined) among the contaminated site fish. However, relative PCB concentration did differ between organs: NB fish (gonad=liver>muscle); I and P fish (gonad>liver>muscle). In contrast to PCB content, PCB congener profiles did show site differences; NB mummichog being depleted in the less chlorinated congeners relative to I and P fish, likely reflecting different PCB sources to these populations. Within a site, however, PCB congener patterns were similar between liver, gonad and muscle. In conclusion, PCA and GLM analyses gave complementary results, both analyses indicating differences in site, but not tissue, distributions of PCB congeners. This study also demonstrates that unlike congener profiles, total PCB content does differ dramatically amongst tissues and further, that PCB differences among tissues (gonad vs. liver vs. muscle) can vary with site. PMID- 12738291 TI - Oxalate production by wood-rotting fungi growing in toxic metal-amended medium. AB - In this report, we have identified oxalic acid as an important metabolite elaborated in the response of wood-rotting fungi to toxic metal stress. The formation of oxalate crystals by white rot fungi (Bjerkandera fumosa, Phlebia radiata and Trametes versicolor) and the brown rot fungus Fomitopsis pinicola, grown on media containing high levels of toxic metal ions has been visualized using scanning electron microscopy (SEM) with energy-dispersive X-ray micro analysis (EDXA) and HPLC. There were no significant differences between the growth of controls (metal-free) and on the 0.5% CaCO(3), Co(3)(PO(4))(2) or Zn(3)(PO(4))(2)-amended plates. ZnO inhibited the growth of all strains. Crystals were not detected in Zn(3)(PO(4))(2)-amended plates. The four examined strains displayed the formation of crystals on ZnO, Co(3)(PO(4))(2) and CaCO(3)-amended plates. PMID- 12738292 TI - Release of substituents from phenolic compounds during oxidative coupling reactions. AB - Phenolic compounds originating from plant residue decomposition or microbial metabolism form humic-like polymers during oxidative coupling reactions mediated by various phenoloxidases or metal oxides. Xenobiotic phenols participating in these reactions undergo either polymerization or binding to soil organic matter. Another effect of oxidative coupling is dehalogenation, decarboxylation or demethoxylation of the substrates. To investigate these phenomena, several naturally occurring and xenobiotic phenols were incubated with various phenoloxidases (peroxidase, laccase, tyrosinase) or with birnessite (delta MnO(2)), and monitored for chloride release, CO(2) evolution, and methanol or methane production. The release of chloride ions during polymerization and binding ranged between 0.2% and 41.4%. Using the test compounds labeled with 14C in three different locations (carboxyl group, aromatic ring, or aliphatic chain), it was demonstrated that 14CO(2) evolution was mainly associated with the release of carboxyl groups (17.8-54.8% of the initial radioactivity). Little mineralization of 14C-labeled aromatic rings or aliphatic carbons occurred in catechol, ferulic or p-coumaric acids (0.1-0.7%). Demethoxylation ranged from 0.5% to 13.9% for 2,6-dimethoxyphenol and syringic acid, respectively. Methylphenols showed no demethylation. In conclusion, dehalogenation, decarboxylation and demethoxylation of phenolic substrates appear to be controlled by a common mechanism, in which various substituents are released if they are attached to carbon atoms involved in coupling. Electron-withdrawing substituents, such as -COOH and -Cl, are more susceptible to release than electron-donating ones, such as -OCH(3) and -CH(3). The release of organic substituents during polymerization and binding of phenols may add to CO(2) production in soil. PMID- 12738293 TI - Variation of stabilised, microbial and biologically active carbon and nitrogen in soil under contrasting land use and agricultural management practices. AB - Land use and agricultural practices modify both the amounts and properties of C and N in soil organic matter. In order to evaluate land use and management dependent modifications of stable and labile C and N soil pools, (i). organic C and total N content, (ii). microbial (C(mic)) and N (N(mic)) content and (iii). C and N mineralisation rates, termed biologically active C and N, were estimated in arable, grassland and forest soils from northern and southern Germany. The C/N ratios were calculated for the three levels (i)-(iii) and linked to the eco physiological quotients of biotic-fixed C and N (C(mic)/C(org), N(mic)/N(t)) and biomass-specific C and N mineralisation rate (qCO(2), qN(min)). Correlations could mainly be determined between organic C, total N, C(mic), N(mic) and C mineralisation for the broader data set of the land use systems. Generally, the mineralisation activity rate at 22 degrees C was highly variable and ranged between 0.11 and 17.67 microg CO(2)-C g(-1) soil h(-1) and -0.12 and 3.81 microg (deltaNH(4)(+)+deltaNO(3)(-))-N g(-1) soil h(-1). Negative N data may be derived from both N immobilisation and N volatilisation during the experiments. The ratio between C and N mineralisation rate differed significantly between the soils ranging from 5 to 37, and was not correlated to the soil C/N ratio and C(mic)/N(mic) ratio. The C/N ratio in the 'biologically active' pool was significantly smaller in soils under conventional farming than those under organic farming systems. In a beech forest, it increased from the L, Of to the Ah horizon. The biologically active C and N pools refer to the current microbial eco physiology and are related to the need for being C and N use efficient as indicated by metabolic qCO(2) and qN(min) quotients. PMID- 12738294 TI - Effects of several low-molecular weight organic acids and phosphate on the adsorption of acid phosphatase by soil colloids and minerals. AB - Adsorption of acid phosphatase on goethite, kaolinite and two colloids from the soils in central and south China in the presence of organic acids and phosphate was studied. With the increase of anion concentration, the ability in decreasing enzyme adsorption followed the sequence: phosphate>tartrate>oxalate>acetate. Acetate showed promotive effect on enzyme adsorption at lower anion concentrations whereas oxalate, tartrate and phosphate compete effectively with enzyme in a broad range of anion concentration. The adsorption isotherms of enzyme in most of the anionic systems studied conformed to the Langmuir equation. Phosphate reduced the affinity of enzyme on goethite more significantly than the other anions. However, tartrate decreased the affinity of enzyme on soil colloids and kaolinite to a greater extent than phosphate, oxalate and acetate. This observation suggested that the impact of anions on enzyme adsorption varies with anionic type and the surface characteristics of soil components. The influence of the addition order of ligand on enzyme adsorption was found greater in tartrate and phosphate systems. In general, simultaneous introduction of ligand and enzyme into the system had the lowest enzyme adsorption, showing more competition between ligand and enzyme molecules in this system. Data from this work indicated that the status and activity of enzyme in certain soil microenvironments especially the rhizosphere where various organic and inorganic ligands are active can be altered and may be completely different from the bulk soil. PMID- 12738295 TI - The dynamics of exchangeable cations in the environment of soils at Kampinoski National Park. AB - A set of physico-chemical properties of soils: soil pH, hydrolytic acidity, alkaline exchangeable cations, cation exchangeable capacity (CEC), and base saturation were studied in six-year long investigations of ecto-humus (organic layer) and endo-humus (Ah horizon) horizons of forest soils at the Kampinoski National Park in Poland. The soil properties determined in the present study showed differentiated values, depending on the actual horizon, the type and degree of soil development advancement, the genesis of the soil parent material (bedrock) as well as on the development of plant community prevailing in given site. PMID- 12738297 TI - Accumulation of nutrients in soils affected by perennial colonies of piscivorous birds with reference to biogeochemical cycles of elements. AB - The accumulation of selected N, K, and P forms in soils within three perennial colonies of black cormorant (Phalacrocorax carbo sinensis) and grey heron (Ardea cinerea) located in northern and eastern Poland were investigated. Soil samples were collected beneath the nests from the most representative for each colony plots. Control samples were taken outside the colonies within sites adjacent to the nesting areas but not affected by bird excrement. From each genetic horizon (20 horizons) in soil profiles, a cumulative sample of about 25-30 kg of soil was taken for analysis. Nitrogen by Kjeldahl, ammonium ions (N(NH(4))), nitrates (N(NO(3))), exchangeable potassium (K(ex)), available potassium (K(av)), and available phosphorus (P(av)) were determined. The soils affected by birds demonstrated a very strong enrichment with N, K, and P in comparison to the control sites, especially in the topsoil horizons. The content of N(NH(4)) in individual soil horizons from the colonies was from 1.7 to 10.1 times higher than the respective control, N(NO(3)) from 2.9 to 215.7, K(ex) from 2.0 to 35.1, K(av) from 1.1 to 48.1, and P(av) in the range from 2.4 to 53.0 times. The highest increment of chemical elements was noticeable in the soils of territories inhabited by cormorants and the least in forest occupied by herons. Some relationships between soil texture and accumulation of biogenic nutrients were determined. Clay loam soil showed the greatest enrichment with analysed forms of elements with the exception of N(NH(4)). PMID- 12738296 TI - Forms and distribution of selenium at different depths and among particle size fractions of three Taiwan soils. AB - The bioavailability of selenium in soils for plants depends more on its forms than on its total concentration. The purpose of the present study was to examine the solid-phase forms of selenium at different depths of three soil series representing major farming soil groups in Taiwan as well as the amounts of selenium in sand, silt and clay fractions of the soils. The study was conducted by means of sequential extraction to obtain the amounts of selenium and the distribution of various solid-phase forms of selenium at different depths of Pinchen (121 degrees 11(')E, 24 degrees 55(')N), Toulun-Sheto (120 degrees 55(')E, 24 degrees 50(')N), and Chunliao (120 degrees 25(')E, 23 degrees 57(')N) soil series. The amounts of metal oxide-bound form of selenium in the three soil series were the largest, with those of Pinchen and Toulun-Sheto soil series exceeding 50% of the total amounts of selenium and that of Chunliao soil series maintained at 30-40%. In the Pinchen and Toulun-Sheto soil series, the amounts of selenium in clay fractions were the largest, with a significant difference between the clays with and without metal oxides and organic matter removed. The amounts of selenium remained high in silt and/or sand fractions of the Chunliao soil series with metal oxides and organic matter removed. Metal oxide and organic matter contents of the three soil series mainly affect the amounts of various solid-phase forms of selenium and their distribution in different depths and particle size fractions of the soils. This observation of selenium associated with soil constituents was in good agreement with the results of the adsorption of selenite and selenate by the three soil series. PMID- 12738298 TI - Seasonal changes of CO(2), CH(4) and N(2)O fluxes in relation to land-use change in tropical peatlands located in coastal area of South Kalimantan. AB - Tropical peatland could be a source of greenhouse gases emission because it contains large amounts of soil carbon and nitrogen. However these emissions are strongly influenced by soil moisture conditions. Tropical climate is characterized typically by wet and dry seasons. Seasonal changes in the emission of carbon dioxide (CO(2)), methane (CH(4)) and nitrous oxide (N(2)O) were investigated over a year at three sites (secondary forest, paddy field and upland field) in the tropical peatland in South Kalimantan, Indonesia. The amount of these gases emitted from the fields varied widely according to the seasonal pattern of precipitation, especially methane emission rates were positively correlated with precipitation. Converting from secondary forest peatland to paddy field tended to increase annual emissions of CO(2) and CH(4) to the atmosphere (from 1.2 to 1.5 kg CO(2)-C m(-2)y(-1) and from 1.2 to 1.9 g CH(4)-C m(-2)y(-1)), while changing land-use from secondary forest to upland tended to decrease these gases emissions (from 1.2 to 1.0 kg CO(2)-C m(-2)y(-1) and from 1.2 to 0.6 g CH(4)-C m(-2)y(-1)), but no clear trend was observed for N(2)O which kept negative value as annual rates at three sites. PMID- 12738299 TI - Fluxes of methane, carbon dioxide and nitrous oxide in boreal lakes and potential anthropogenic effects on the aquatic greenhouse gas emissions. AB - We have examined how some major catchment disturbances may affect the aquatic greenhouse gas fluxes in the boreal zone, using gas flux data from studies made in 1994-1999 in the pelagic regions of seven lakes and two reservoirs in Finland. The highest pelagic seasonal average methane (CH(4)) emissions were up to 12 mmol x m(-2) x d(-1) from eutrophied lakes with agricultural catchments. Nutrient loading increases autochthonous primary production in lakes, promoting oxygen consumption and anaerobic decomposition in the sediments and this can lead to increased CH(4) release from lakes to the atmosphere. The carbon dioxide (CO(2)) fluxes were higher from reservoirs and lakes whose catchment areas were rich in peatlands or managed forests, and from eutrophied lakes in comparison to oligotrophic and mesotrophic sites. However, all these sites were net sources of CO(2) to the atmosphere. The pelagic CH(4) emissions were generally lower than those from the littoral zone. The fluxes of nitrous oxide (N(2)O) were negligible in the pelagic regions, apparently due to low nitrate inputs and/or low nitrification activity. However, the littoral zone, acting as a buffer for leached nitrogen, did release N(2)O. Anthropogenic disturbances of boreal lakes, such as increasing eutrophication, can change the aquatic greenhouse gas balance, but also the gas exchange in the littoral zone should be included in any assessment of the overall effect. It seems that autochthonous and allochthonous carbon sources, which contribute to the CH(4) and CO(2) production in lakes, also have importance in the greenhouse gas emissions from reservoirs. PMID- 12738300 TI - Arsenic behavior in newly drilled wells. AB - In the present paper, inorganic arsenic species and chemical parameters in groundwater were determined to investigate the factors related to the distribution of arsenic species and their dissolution from rock into groundwater. For the study, groundwater and core samples were taken at different depths of two newly drilled wells in Huron and Lapeer Counties, Michigan. Results show that total arsenic concentrations in the core samples varied, ranging from 0.8 to 70.7 mg/kg. Iron concentration in rock was about 1800 times higher than that of arsenic, and there was no correlation between arsenic and iron occurrences in the rock samples. Arsenic concentrations in groundwater ranged from <1 to 171 microg/l. The arsenic concentration in groundwater depended on the amount of arsenic in aquifer rocks, and as well decreased with increasing depth. Over 90% of arsenic existed in the form of As(III), implying that the groundwater systems were in the reduced condition. The results such as high ferrous ion, low redox potential and low dissolved oxygen supported the observed arsenic species distribution. There was no noticeable difference in the total arsenic concentration and arsenic species ratio between unfiltered and filtered (0.45 microm) waters, indicating that the particulate form of arsenic was negligible in the groundwater samples. There were correlations between water sampling depth and chemical parameters, and between arsenic concentration and chemical parameters, however, the trends were not always consistent in both wells. PMID- 12738301 TI - Total mercury concentrations in an industrialized catchment, the Thur River basin (north-eastern France): geochemical background level and contamination factors. AB - River bottom sediments and soils were collected from the industrialized Thur River basin (north-eastern France) to assess mercury contamination. The regional geochemical background level of total mercury was evaluated to calculate mercury contamination factors (Fc) in soils and river bottom sediments. Our estimate of the mean background mercury levels in river sediments and soils, not affected by human activities, was 232 ng x g(-1) (range: 27-406 ng x g(-1)). Sediments contaminated by the effluent from a chlor-alkali plant yielded the highest contamination factors (Fc=1784). Contamination factors of surficial soils within 1 km of the industrial site range from 6.3 to 43.6. This contamination is attributed to diffuse atmospheric deposition from this local plant. However, even upstream from this industrial area elevated contamination factors were recorded for river bottom sediments (Fc=3.2 to 26.4) and for one alluvial soil profile (Fc=10). This is possibly due to past pollution resulting from waste water discharges. Mercury contamination in the different horizons of alluvial soils is not correlated with soil organic carbon content, but may be the result of occasional accidental pollution arising from the introduction of contaminated suspended particulate matter by the Thur River during periods of flooding. PMID- 12738302 TI - Horizontal and vertical variabilities of mercury concentration and speciation in sediments of the Gdansk Basin, Southern Baltic Sea. AB - The mercury compounds introduced to the environment because of anthropogenic activity are accumulated, mainly, in marine sediments. Both distribution of mercury in the Baltic Sea and factors affecting it are remain largely unknown. Due to its complex chemistry and variable conditions in the Baltic Sea bottom sediments, mercury may be reemitted to the overlaying water, and thus to the environment. The aim of this study was twofold. Total mercury contents were measured in sediments of the Gdansk Basin along a Vistula mouth (main source)- Gdansk Deep (deposition area) transect in order to assess spatial distribution of the element. Soft and sandy bottom sediment cores were collected and cut into slices. The mercury measurements consisted of acid digestion followed by CV-AAS determination. A five-step sequential extraction procedure involving measurements of mercury species water soluble, bound to humic substances and insoluble, was employed in order to investigate mercury speciation. On the basis of speciation results stability and remobilisation potential was assessed. The second aim was demonstrating that mercury gradients in vertical profiles of the investigated sediment cores exist. Total mercury contents varied in range from 28 ng/g x dw to 844 ng/g. Mercury associated with organic matter and sulphides, (respectively 39% and 49%) were the dominant species of the element. The results indicated that the river run-off is a source of mercury in the area. Vertical gradients of both total content and speciation of mercury in the soft sediments were attributed to both remobilisation from sediments, and decreasing trend in the anthropogenic load of the element deposited to sediments. PMID- 12738303 TI - 2002 Radiation Oncology Workforce Study: American Society for Therapeutic Radiology and Oncology. PMID- 12738304 TI - Infrastructure of radiotherapy in Spain: a minimal standard of radiotherapy resources. AB - PURPOSE: To assess the state of our specialty, the Spanish Society of Radiotherapy and Oncology ordered a survey of all Spanish services of radiation oncology. METHODS AND MATERIALS: In June 1999, the Society ordered an analysis of the state of radiation oncology. It created a survey that was sent to all radiotherapy units in Spain. A database was created in which 230 variables were analyzed. RESULTS: Eighty-four centers were analyzed, and 157 external beam irradiation, megavoltage units were counted, of which 67 were cobalt units and 90 were linear accelerators. The cobalt units worked an average of 11.4 h daily and the linear accelerators 11.6 h. The number of patients/unit/y was 472 for the cobalt units and 442 for the linear accelerators. The number of patients by physician and year was 179. Each center received a mean of 958 new patients annually. The average between the reception and start of treatment was 25.52 days (maximum 60), and it was estimated that only 38.1% of cancers were irradiated. The number of radiation oncologists working was 392. Spain has a deficit of 297 radiation oncologists. CONCLUSION: There is a need for 44 MV units and for the replacement of 67 cobalt units. The present lack of units has had an impact on palliative treatment, which has resulted in pharmacy costs. As long as these instrumental deficiencies are not solved, waiting lists will continue to be inherent to the system. There are also important staff deficiencies, in that about 297 radiation oncologists would be needed to cover the needs. PMID- 12738306 TI - Stereotactic single-dose radiotherapy of stage I non-small-cell lung cancer (NSCLC). AB - PURPOSE: The treatment of early-stage lung cancers is a primary domain of thoracic surgery, leading to persuasive results. In patients with medical contraindications, radiotherapy is an alternative, although with considerably worse outcome. Radiotherapy is associated with the risk of severe acute side effects and a permanent decrease of lung function. By the introduction of an extracranial stereotactic treatment technique, the amount of normal tissue in the high-dose region can be reduced, allowing the performance of single-dose treatment with high, biologically effective doses. METHODS AND MATERIALS: Between October 1998 and May 2001, 10 patients with histologically confirmed Stage I non small-cell lung cancer were treated with stereotactic single-dose radiotherapy. A self-developed stereotactic frame was used for patient positioning and navigation. Total doses applied ranged from 19 to 26 Gy. After treatment, regular CT-based follow-up was performed. RESULTS: During a median follow-up period of 14.9 months, the tumors in 8 of 10 patients were locally controlled. The actuarial overall survival was 80% and 64%, respectively, 12 and 24 months after therapy. Actuarial local recurrence-free survival reached 88.9% and 71.1%, respectively. Therapy-related perifocal normal-tissue reaction occurred in 70% of all treated patients, although no major clinical symptoms were seen. In 5 patients, systemic metastases were found during follow-up; 1 patient developed suspect mediastinal lymph nodes. CONCLUSION: Stereotactic single-fraction radiotherapy is a feasible, safe, and effective procedure for the treatment of Stage I non-small-cell lung cancer. It promises high local control with a reduced overall treatment time. However, further investigation in a larger patient collective with extended follow-up is necessary. PMID- 12738305 TI - Mature survival results with preoperative cisplatin, protracted infusion 5 fluorouracil, and 44-Gy radiotherapy for esophageal cancer. AB - PURPOSE: To assess the long-term survival results after cisplatin, protracted infusion 5-fluorouracil, and concurrent radiotherapy (RT) followed by surgical resection of esophageal cancer. METHODS AND MATERIALS: Ninety-two patients with esophageal cancer (65 with adenocarcinoma and 27 with squamous cell carcinoma) were treated in two sequential protocols of preoperative chemoradiotherapy. The patients had tumor confined to the esophagus and regional nodes, including celiac nodes for middle and distal lesions. In trial A (1989-1994), 50 patients were treated with 44 Gy RT (2 Gy/d) along with concurrent 5-fluorouracil 300 mg/m(2)/d given by protracted venous infusion on Days 1-30 and cisplatin 26 mg/m(2) on Days 1-5 and 26-30. In trial B (1995-1997, 42 patients), the chemotherapy dosages during RT were reduced to 5-fluorouracil 225 mg/m(2)/d protracted venous infusion and cisplatin 20 mg/m(2)/d on Days 1-5 and 16-30; three cycles of paclitaxel 135 mg/m(2)and cisplatin 75 mg/m(2) were given postoperatively. Surgery generally occurred 4-6 weeks after completion of the planned preoperative therapy. Transhiatal resection was performed whenever possible. RESULTS: Of the 92 patients, 86 (93%) underwent surgery (1 refused, 2 died preoperatively, and 3 developed evidence of metastatic disease). Of the 92 patients, 80 (87%) had complete resections with negative margins (3 had positive margins and 3 had distant metastases discovered at surgery). The pathologic complete response rate was 33% (30 of 92). The median follow-up was 63.5 months. The median survival and disease-specific survival for all enrolled patients was 35 and 59 months, respectively. The 5-year survival and disease-specific survival rate was 40% and 49%, respectively. Patients with a pathologic complete response had a 67% survival rate at 5 years (median not reached), and the remainder of patients had a 5-year survival rate of 27% (median 21 months; p <0.001). For 21 patients alive after 5 years (60-121 months), 2 died of their disease and all others were disease free. Eight patients with pathologic Stage I tumor at the time of surgery had survival similar to those with a complete response to preoperative therapy. The median survival for patients with pathologic Stage IIA, IIB, III, and IV disease at the time of surgery was 22, 13.5, 18, and 4.9 months, respectively. The pattern of initial failure was local/regional alone in 6% (5 of 90), local/regional plus distant in 3% (3 of 90), and distant alone in 47% (42 of 90). No differences were noted in survival or response rate between those with adenocarcinoma or squamous cell carcinoma. CONCLUSION: The promising 5-year survival results and low rate of late cancer-related deaths suggest that these regimens of intensive neoadjuvant therapy may improve the overall cure rate. The pathologic stage after neoadjuvant therapy is an important predictor of survival and may be useful in selecting patients for novel adjuvant therapies. Isolated local failure is uncommon, indicating that efforts to improve the therapeutic outcome should focus on optimizing systemic therapy rather than intensifying the RT. Additional randomized data are needed to assess the benefits of this therapeutic approach fully. PMID- 12738307 TI - Elective nodal failures are uncommon in medically inoperable patients with Stage I non-small-cell lung carcinoma treated with limited radiotherapy fields. AB - PURPOSE: To review the outcome for 56 Stage I non-small-cell lung cancer treated definitively with three-dimensional conformal radiotherapy (3D-CRT) and to investigate the value of elective nodal irradiation in this patient population. METHODS AND MATERIALS: Between 1992 and 2001, 56 patients were treated with 3D CRT for inoperable Stage I histologically confirmed non-small-cell lung cancer; 31 with T1N0 and 25 with T2N0 disease. All patients were treated with 3D-CRT to a median isocenter dose of 70 Gy (range 59.94-83.85) given in daily doses of 1.8 or 2 Gy. Prognostic factors were analyzed with respect to their impact on overall survival. Twenty-two patients received radiotherapy (RT) directed to elective regional lymphatics to doses of 45-50 Gy. The remaining 33 patients were treated to limited fields confined to the primary lung cancer with a margin. The patterns of failure were reviewed. RESULTS: The median follow-up was 20 months (range 6 months to 6 years). The actuarial local control rate was 88%, 69%, and 63%, at 1, 2, and 3 years, respectively. The actuarial cause-specific survival rate was 82%, 67%, and 51% at 1, 2, and 3 years, respectively. The actuarial overall survival rate was 73%, 51%, and 34% at 1, 2, and 3 years, respectively. The actuarial metastasis-free survival rate was 90%, 85%, and 81% at 1, 2, and 3 years, respectively. The RT dose was the only factor predictive of overall survival in our analysis. No statistically significant difference was noted in cause-specific or overall survival according to whether patients received elective nodal irradiation. Two of 33 patients treated with limited fields had regional nodal failure. CONCLUSION: Many patients with medically inoperable Stage I lung cancer die of intercurrent causes. The omission of the elective nodal regions from the RT portals did not compromise either the cause-specific or overall survival rate. Elective nodal failures were uncommon in the group treated with limited RT fields. A radiation dose 70 Gy was predictive of better survival in our population. We await the results of prospective trials evaluating high-dose RT in patients treated with RT alone for Stage I lung cancer. PMID- 12738308 TI - Tumor location cannot predict the mobility of lung tumors: a 3D analysis of data generated from multiple CT scans. AB - PURPOSE: There is limited information available on the three-dimensional (3D) motion of lung tumors. Data derived from multiple planning computed tomographic (CT) scans were used to characterize the 3D movement of small peripheral lung tumors. METHODS AND MATERIALS: A total of 29 data sets from patients with Stage I non-small-cell lung cancer (NSCLC), each of which consisted of three "rapid" and three "slow" planning CT scans, were analyzed. All six scans were coregistered, and contoured gross tumor volumes (GTVs) were expanded by 5 mm to derive clinical target volumes (CTVs). Two-dimensional and 3D displacement vectors of the individual CTVs, relative to an "optimal" CTV derived from all six scans, were generated. Tumor mobility was correlated with location. Three-dimensional margins, which had to be added to individual CTVs to ensure coverage of "optimal" CTVs, were determined. RESULTS: No significant correlation was observed between the anatomic location of tumors and the extent of mobility in the x, y, and z axes. However, supradiaphragmatic lesions exhibited more mobility, particularly in the craniocaudal direction. The addition of a 3D margin of 5 mm to a single slow CTV ensured full coverage of the "optimal CTV". CONCLUSIONS: Lung tumors demonstrate significant mobility in all directions, and this did not closely correlate with anatomic location. Individualized assessment of tumor mobility remains necessary, and is possible when the CTV derived from a single slow scan is used for radiotherapy planning. PMID- 12738309 TI - Routine use of approximately 60 Gy once-daily thoracic irradiation for patients with limited-stage small-cell lung cancer. AB - PURPOSE: To review the outcome of patients with limited-stage small-cell lung cancer receiving daily thoracic irradiation (RT) to approximately 60 Gy. METHODS AND MATERIALS: The records of patients treated with RT for limited-stage small cell lung cancer between 1991 and 1999 at Duke University were retrospectively reviewed. Sixty-five patients were identified who had received continuous course once-daily 1.8-2 Gy fractions to approximately 60 Gy (range 58-66). All patients received chemotherapy (CHT); 32 received concurrent RT/CHT and 33 sequential CHT and then RT. Prophylactic cranial RT was administered to 17 patients. The time from diagnosis to local failure, tumor progression, and death was assessed using actuarial methods. The median follow-up for all patients was 16.7 months and for surviving patients was 29.6 months. The median age was 64 years (range 36-83), and the median Karnofsky performance status was 80 (range 50-100). RESULTS: The 3 year actuarial rate of local failure, progression-free survival, and overall survival was 40%, 25%, and 23%, respectively. One case of acute Grade 3 esophagitis developed. Ten late complications occurred: four pulmonary, two esophageal, two infectious, one leukemia, and one retinal toxicity with prophylactic cranial RT. Six were mild and resolved with treatment. CONCLUSION: CHT plus approximately 60 Gy of once-daily RT for limited-stage small-cell lung cancer was generally well tolerated. The survival rates were less than have been reported using 45 Gy in 1.5-Gy twice-daily fractions (2-year overall survival rate 47% compared with 30% in this study), but may be comparable because fewer than one-half our patients received concurrent CHT/RT and only 26% received prophylactic cranial RT. The relatively low rate of normal tissue morbidity in our patients supports the use of conventional once-daily fractionation to > or = 60 Gy. A randomized trial would be required to compare the outcomes after maximally tolerated dose twice-daily RT vs. maximally tolerated dose daily RT. PMID- 12738310 TI - Radiation pneumonitis following treatment of non-small-cell lung cancer with continuous hyperfractionated accelerated radiotherapy (CHART). AB - PURPOSE: To determine whether partial volume lung irradiation influences the risk of developing acute radiation pneumonitis after the treatment of non-small-cell lung cancer with continuous hyperfractionated accelerated radiotherapy (CHART). METHODS AND MATERIALS: We conducted an analysis of 32 patients treated with CHART at the Gloucestershire Oncology Center. Twelve patients were treated using conventional two-dimensional treatment techniques and received elective nodal irradiation (ENI). Their treatment plans were subsequently recapitulated using a three-dimensional treatment planning system. Twenty patients were planned using this system from the outset. For these patients, elective nodal irradiation was omitted. Dose-volume histograms (DVH) were constructed and several parameters analyzed for their ability to predict for the development of pneumonitis. RESULTS: Univariate analysis revealed that the percentage lung volume receiving more than 20 Gy (V20) and the mean lung dose are of predictive value for the development of pneumonitis after CHART. There is a strong correlation between these two parameters. Importantly, partial volume lung irradiation using CHART appears to be better tolerated than conventionally fractionated radiotherapy. The omission of ENI considerably reduces V20. Using a commonly employed 3-beam technique it was also noted that the shape of the planning target volume (PTV) in the transverse plane (expressed as an elliptical index) affects the conformity of the V20 isodose to the PTV. This influences the scope for dose escalation with irregularly shaped tumors. CONCLUSIONS: In relation to acute radiation pneumonitis, CHART appears to have a superior therapeutic index than conventionally fractionated radiotherapy. V20 and mean lung dose are useful factors for predicting the risk of this complication. The use of these parameters will aid the selection of optimal treatment plans and provides a basis for future dose escalation studies. PMID- 12738311 TI - Can 99mTc-DTPA aerosol scanning predict radiation pneumonitis? AB - PURPOSE: To investigate whether changes in diethylenetriamine pentaacetic acid (DTPA) aerosol clearance lung scans predict the development of radiation pneumonitis. METHODS AND MATERIALS: Thirty-three patients with advanced non-small cell lung cancer were treated with palliative radiation therapy to thorax. All patients were subjected to pretreatment and post-treatment DTPA aerosol clearance lung scans. The clearance t1/2 values were compared using the paired t test. Changes in percentage aerosol deposition were also compared. The patients were serially assessed clinically and radiologically for development of radiation pneumonitis. RESULTS: For the whole group, the mean DTPA clearance t1/2 for the diseased lung fell from 36.33 to 28.85 min (p = 0.17). Twelve patients developed radiation pneumonitis, 8 Grade 1, 2 Grade 2, 1 Grade 3, and 1 Grade 4. In patients who developed radiation pneumonitis, the clearance t1/2 for the diseased lung decreased from 37.50 min to 29.00 min after treatment (p = 0.50). Other subgroups analyzed, including smokers, nonsmokers, those with and without endobronchial disease, as well as those free from radiation pneumonitis, also showed no significant change in the clearance of the aerosol. The difference between the percentage aerosol deposition of the diseased lung (37.92%) and that of the opposite lung (62.08%) for the entire sample was significantly different both before and after treatment (p < 0.01). Delivery of radiation did not significantly change the aerosol deposition in either lung. CONCLUSIONS: We conclude that, in patients with advanced non-small-cell lung cancer, significant change in the pulmonary clearance of 99mTc-DTPA aerosol could not be demonstrated after radiation therapy. In such patients, clearance of the aerosol does not predict the development of radiation pneumonitis. PMID- 12738313 TI - Intrafraction motions of the larynx during radiotherapy. AB - PURPOSE: To quantify the intrafraction motions of the larynx during radiotherapy of laryngeal cancer. Depending on the magnitude, duration, and incidence, these motions may have clinical consequences for the choice of margins around the clinical target volume. METHODS AND MATERIALS: The intrafraction motions were analyzed for 10 patients, treated in 33 or 35 fractions. The intrafraction motions of the larynx were visualized using an a-Si flat panel imager. Images were obtained every 200 ms, resulting in a movie of images for each beam. In addition to visual analysis of all movies, the tip of the epiglottis was delineated and used as a landmark, the coordinates of which were followed in time. RESULTS: Movies were obtained during 79% of the total number of radiotherapy fractions. The total duration of swallowing was on average 0.45% (range 0.0-1.5) of the total irradiation time. Deviations of motions other than swallowing ranged between 0.3 and 11.5 mm. Some of these motions were more sudden, and others were probably related to breathing, because the frequency of these motions was 8-20/min. CONCLUSION: The incidence and total duration of swallowing is low. Therefore, it is not necessary to apply an internal margin to take into account these displacements. Other motions, however, occur more often. In 95% of the irradiation time, the tip of the epiglottis moves within a range of 7.1 mm. A margin should be applied to the clinical target volume to take into account these motions. PMID- 12738312 TI - Comparison of the comet assay and the oxygen microelectrode for measuring tumor oxygenation in head-and-neck cancer patients. AB - PURPOSE: To compare the Eppendorf PO2 histograph and the alkaline comet assay as methods of measuring tumor hypoxia in patients with head-and-neck squamous cell carcinomas. MATERIALS AND METHODS: As part of a larger clinical trial, 65 patients with head-and-neck squamous cell carcinoma nodal metastasis underwent tumor oxygenation measurements with Eppendorf PO2 histographs and comet assays, performed on fine-needle aspirates at 1 and 2 min after 5 Gy. Fifty-four patients had sufficient tumor cells for comet analysis at 1 min and 26 at both 1 and 2 min. Individual cells were examined for DNA single-strand breaks by alkaline gel electrophoresis, and the distribution of values was quantified using median tail moment (MTM). Nonirradiated tumor cells from pretreatment fine-needle aspirates received 5 Gy in vitro to establish the oxygenated response. RESULTS: There was a significant correlation between the 1- and 2-min MTM (slope = 0.77 +/- 0.03). There was no relationship between DNA damage in tumor cells irradiated in vitro and in vivo. No correlation was found between Eppendorf PO2 measurements and comet MTM. There was a statistically significant correlation between the treatment response in the node studied and comet MTMs, whereas no correlation was observed between treatment response and Eppendorf measurements. CONCLUSIONS: Comet assays are reproducible, as shown by biopsies at 1 and 2 min. Intertumor variation in the MTM is not a result of intrinsic radiosensitivity but of tumor hypoxia. There was no correlation between Eppendorf PO2 measurements and comet MTM. Comet assays were better than Eppendorf in predicting treatment response as an end point for short-term outcome. Longer follow-up is needed to determine the role of the comet assay as a predictor for locoregional tumor control and survivals. PMID- 12738314 TI - Feasibility and toxicity of combined photon and carbon ion radiotherapy for locally advanced adenoid cystic carcinomas. AB - PURPOSE: To investigate clinical feasibility and toxicity of combined photon and carbon ion radiotherapy in locally advanced adenoid cystic carcinomas (ACC) within a prospective Phase I/II trial. METHODS AND MATERIALS: Between September 1998 and April 2002, 16 patients with histopathologically proven ACC and residual macroscopic tumor were treated with combined photon RT and a carbon ion boost to the macroscopic tumor. Median total tumor dose within the gross tumor volume (GTV) was 72 GyE. Photon radiation therapy (RT) consisted of fractionated stereotactic RT in 7 patients; 9 patients received stereotactic intensity modulated RT. Carbon ion boost was delivered by intensity-controlled raster scanning at the heavy ion synchrotron (SIS) at the Heavy Ion Research Center (GSI) in Darmstadt. RESULTS: Median follow-up was 12 months. Three patients developed locoregional recurrences 9, 11, and 24 months after RT, respectively. Actuarial local control rates were 80.8% and 64.6% at 1 and 3 years, respectively. Overall survival rates were 100% and 83.3% at 1 and 3 years, respectively. Acute side effects greater than Common Toxicity Criteria (CTC) Grade 2 were observed in 2 patients; no patient developed late effects > CTC Grade 2. CONCLUSIONS: Combined photon and carbon ion RT is feasible and effective in patients with locally advanced ACC. Acute and late toxicity is moderate with respect to the delivered tumor doses and in accordance with the radiobiologic modeling. A Phase III trial is designed. PMID- 12738315 TI - Time factor in postoperative radiotherapy: a multivariate locoregional control analysis in 868 patients. AB - PURPOSE: To study locoregional tumor control in postoperative radiotherapy (PRT) for head-and-neck cancer in relation to the position and duration of treatment gaps, duration of the interval surgery-radiotherapy, and to the other potentially prognostic variables. METHODS AND MATERIALS: The retrospective study included 868 patients with cancer of the larynx, oral cavity, oropharynx, and hypopharynx treated in Gliwice between 1980 and 1997. Mean total radiation dose, dose per fraction, overall radiation treatment time (OTT), and the interval surgery-PRT were 62.8 Gy, 2.1 Gy, 45 days, and 63 days, respectively. No interruptions during PRT (except for weekend breaks) appeared in 30% of patients, whereas 19% had more than 5 days of gap. The actuarial locoregional recurrence-free survival (RFS) has been examined using multivariate Cox regression model, and the ultimate locoregional tumor control probability using a logistic model. RESULTS: Increased duration of treatment gaps, positive resection margins, pathologically proven metastases to the neck nodes, and extralaryngeal site of cancer were significantly related to a decrease in RFS. The duration of time interval surgery PRT appeared, by contrast, only borderline significant for RFS. The relative risk of locoregional recurrence was approximately the same for the gaps in days 1-21 as for the "late" gaps (days >21), except in a subgroup of patients with positive clinical margins. The logistic analysis revealed a significant time-related displacement of the dose-response curves for PRT. The detriment from the protraction of OTT appeared to be larger than the benefit from the equivalent shortening of OTT. CONCLUSIONS: Although the conclusions from this study must be regarded as only hypothesis-generating, we assume that a highly significant adverse influence of radiation treatment gaps on the rate of tumor control is consistent with rapid repopulation of cancer clonogenes during PRT. Lack of significant effect of the position of gaps on locoregional tumor control after radical surgery may suggest that a lag time for the onset of repopulation in PRT is short. A less likely explanation is that the total amount of regeneration during OTT is the same, regardless of the timing of the gap, even if all the repopulation occurred late. The magnitude of the detriment in tumor control from prolonged interval surgery-PRT indicates that repopulation of cancer cells between surgery and radiotherapy is not as fast as between the fractions of radiotherapy. PMID- 12738316 TI - In-depth evaluation of the AJCC/UICC 1997 staging system of nasopharyngeal carcinoma: prognostic homogeneity and proposed refinements. AB - PURPOSE: To critically evaluate the American Joint Commission on Cancer (AJCC)/International Union Against Cancer (UICC) 1997 staging system and look back on its achievements by comparing it with the AJCC/UICC 1992 and Ho 1978 staging systems. To identify areas for additional refinement, we analyzed the prognostic heterogeneity within each stage in depth, which provided important clues for the addition or better categorization of the different defining criteria. METHODS AND MATERIALS: We performed a retrospective review of the data from 1294 consecutive biopsy-proven nonmetastatic nasopharyngeal carcinoma patients and staged the extent of disease according to the defining criteria of the three staging systems. All patients had undergone detailed pretreatment assessment by fiberoptic endoscopy and CT. Radical-intent radiotherapy was given using the Ho technique according to our standard protocol. RESULTS: The AJCC/UICC 1997 staging system was superior to the other two staging systems, because it assigned patients to more uniform-size stage groupings and correlated better with prognosis. Parapharyngeal space involvement was not an independent predictor for survival, local control, or metastasis. On the other hand, carotid space involvement correlated with a greater likelihood of metastasis. Prognostic heterogeneity was found. Those with orbit, cranial nerve, or intracranial involvement fared worse within Stage T4; those with a maximal lymph node size >3 cm fared worse within Stage N2; and those with bilateral lymph node metastasis fared worse within Stage N3. CONCLUSION: The prognostic accuracy of the AJCC/UICC 1997 staging system can be improved further by recategorization of the T, N, and group stage criteria. PMID- 12738317 TI - Second primary cancers in patients with laryngeal cancer: a population-based study. AB - BACKGROUND: Literature regarding incidence of site-specific second cancers after laryngeal cancer is limited. Risk factors associated with second primaries are unknown. METHODS: Second primaries after laryngeal cancer in the SEER database (1973-1996) were analyzed for incidence, relative risk compared with the general population, and potential risk factors, including radiotherapy. Information on chemotherapy and tobacco smoking was not available in the SEER database. RESULTS: Of 20,074 laryngeal cancer patients surviving at least 3 months, 3533 (17.6%) developed second cancers. The cumulative risk of developing a second cancer was 26% at 10 years and 47% at 20 years. Compared with age-adjusted, gender, and tumor-specific rates in the general population, laryngeal cancer patients had higher risks of second cancers overall (observed-to-expected ratio [O/E] = 1.68, 95% confidence interval [CI] = 1.58-1.79), head-and-neck (4.81 [4.31-5.58]), esophageal (3.99 [3.29-4.83]), and lung (3.56 [3.34-3.79]) cancer. Advanced age at initial diagnosis was associated with increased risks of second cancers (p = 0.0001). Radiotherapy was associated with increased risk of second cancers overall (relative risk [RR] = 1.10 [1.02-1.18], p = 0.012), especially second cancers of the lung (RR = 1.18, [1.05-1.33], p = 0.006) and possibly second cancers of the head and neck (RR = 1.26, [0.99-1.60], p = 0.061). Radiotherapy was associated with a 68% excess risk (RR = 1.68, [1.16-2.43], p = 0.007) of developing a second head-and-neck cancer in patients who survived more than 5 years. Second primary was associated with a poor survival (p = 0.0001). CONCLUSIONS: Second cancers after laryngeal cancer are common, especially for long-term survivors. Radiotherapy was associated with a small increased risk of developing second cancers overall and long-term risk of head-and-neck cancers. This data should be interpreted with caution in light of the lack of information on chemotherapy and tobacco smoking in the SEER database. Prevention and early detection are indicated. PMID- 12738318 TI - Experience of ultrasound-based daily prostate localization. AB - PURPOSE: The NOMOS (Sewickley, PA) B-mode Acquisition and Targeting System (BAT) ultrasound system provides a rapid means of correcting for interfraction prostate positional variation. In this investigation, we report our experience on the clinical issues relevant to the daily use of the BAT system and the analysis of combined setup error and organ motion for 3509 BAT alignment procedures in 147 consecutive patients treated with IMRT for prostate cancer. METHODS AND MATERIALS: After setup to external skin marks, therapists performed the BAT ultrasound alignment procedure before each IMRT treatment. In this study, a single physician (A.C.) reviewed all BAT images and classified image quality and accuracy of image alignment by the therapist. On a scale of 1-3, near-perfect image quality or alignment was given a 1, fair image quality or misalignment > or = 5 mm (likely within the PTV) was given a 2, and unacceptable image quality or misalignment >5 mm (potential to violate the PTV) was given a value of 3. The distribution of shifts made was analyzed in each dimension and for all patients. The time required to perform the BAT alignment was also assessed in 17 patients. RESULTS: Among the 3509 attempted BAT procedures, the image quality was judged to be poor or unacceptable in 5.1% (181). Of the remaining 3328 BAT images, with quality scores of 1-2, alignments were unacceptable (>5 mm misalignment as judged by the reviewing physician) in 3% (100). The mean shift in each direction, averaged over all patients, was 0.5-0.7 mm. Interfraction standard deviation (1 SD) of prostate position based on combined setup error and internal organ motion is 4.9 mm, 4.4 mm, and 2.8 mm in the anteroposterior (AP), superior-inferior (SI), and lateral (RL) dimensions, respectively. The distribution of the shifts was a near-random Gaussian-type in all three major axes, with greater variations in AP and SI directions. The percent of BAT procedures in which the shift was >5 mm was 28.6% in AP, 23% in SI, and 9% in RL directions. The average BAT procedure took extra 5 min out of a 20-min time slot in a typical eight-field IMRT treatment. CONCLUSIONS: The quality of the daily ultrasound images was deemed acceptable in 95%. Major alignment errors by therapists were only 3%. The BAT system is clinically effective and feasible in a matter of 5 min. Although the accuracy of the BAT was not addressed in this investigation, we found a significant percentage of large shifts being made from the initial alignment position. PMID- 12738319 TI - Prostate-specific antigen bounce after prostate seed implantation for localized prostate cancer: descriptions and implications. AB - PURPOSE: To calculate the actuarial risk of developing a prostate-specific antigen (PSA) bounce after prostate brachytherapy alone, using three definitions of bounce mentioned in the literature, and to explore the relationship between disease and treatment variables and the risk of developing a bounce. The impact of PSA bounce on PSA failure was also explored. METHODS AND MATERIALS: A total of 373 patients with T1-T2 prostate cancer underwent radioactive seed implant using 125I (n = 337) or 103Pd (n = 36) without hormonal therapy or external beam RT. All patients had a minimum of 1 year (median 4, maximum 11) of follow-up and at least three follow-up PSA values. PSA bounce was defined by a rise of one or two PSA values with a subsequent fall. Three definitions of bounce were used: definition 1, rise > or = 0.1 ng/mL; definition 2, rise > or = 0.4 ng/mL; and definition 3, rise >35% of previous value. RESULTS: The actuarial likelihood of experiencing a PSA bounce at 5 years was 31% for definition 1, 17% for definition 2, and 20% for definition 3. The median time to develop a bounce was 19.5 months for definitions 1 and 2 and 20.5 months for definition 3. Gleason score, initial PSA level, and clinical stage did not predict for bounce using any definition. Using definition 1, younger patients (< or = 65 years) had a bounce rate at 5 years of 38% vs. 24% for older patients (p = 0.009). 125I patients receiving an implant dose of < or = 160 Gy had a bounce rate (definition 1) at 5 years of 24% vs. 38% for those receiving a dose delivered to 90% of the gland on the 1 month postimplant dose-volume histogram (D90) >160 Gy (p = 0.04). Using definition 2, prostate volume significantly affected the incidence of bounce. Patients with larger glands (>35 cm(3)) were more likely to experience a bounce (23% at 5 years) than those with smaller glands (< or = 35 cm(3)) who had a bounce rate of 11% at 5 years (p = 0.01). In a multivariate analysis of factors predicting for PSA failure, PSA bounce was not found to be significant. CONCLUSION: PSA bounce is a common phenomenon after prostate brachytherapy and occurs at a rate of 17 31%, depending on the definition used. It is more common in younger patients, those receiving higher implant doses, and those with larger glands. PSA bounce does not predict for future PSA failure. PMID- 12738320 TI - Long-term urinary quality of life after permanent prostate brachytherapy. AB - PURPOSE: To evaluate late urinary function after permanent prostate brachytherapy using a validated, patient-administered quality-of-life instrument. METHODS AND MATERIALS: A total of 225 consecutive patients underwent prostate brachytherapy between April 1995 and March 1998. Of the 225 patients, 17 had died and 3 had been institutionalized secondary to Alzheimer's disease. Of the remaining 205 patients, each was mailed a self-administered questionnaire (the urinary function component of the Expanded Prostate Cancer Index [EPIC] and the International Prostate Symptom Score [IPSS]). Of the 205 surveys mailed, 195 (95.1%) were returned. The mean and median follow-up was 66.3 and 64.0 months, respectively. The clinical parameters evaluated included age, pretreatment prostate-specific antigen level, Gleason score, stage, risk group, prostate volume, presence of diabetes and hypertension, and tobacco consumption. The treatment parameters included the ultrasound planning volume, hormonal status, use of supplemental external beam radiotherapy, isotope, and follow-up. The dosimetric parameters included values of the minimal dose received by 90% of the prostate gland and the percentage of prostate volume receiving 100%, 150%, and 200% of the prescribed minimal peripheral dose. Because detailed baseline urinary function was not available, a cross-sectional survey was performed in which 51 newly diagnosed prostate cancer patients of comparable demographics served as controls. RESULTS: When the survey scores for the implant patients were compared with the control group, no significant differences in either the IPSS or function, bother, incontinence, or irritation/obstruction subscales of the urinary EPIC were discernible. In addition, no significant difference was observed between the implant and control groups when the EPIC and IPSS surveys were evaluated by each individual question. Of all the evaluated parameters, the use of tobacco was the best predictive variable for diminished quality of life. CONCLUSION: No significant difference was noted in the overall long-term urinary quality of life when brachytherapy patients were compared with a group of newly diagnosed prostate cancer patients of comparable demographics. Of all parameters evaluated, tobacco consumption was the single strongest predictor of late urinary function. PMID- 12738321 TI - Is intraoperative nomogram-based overplanning of prostate implants necessary? AB - PURPOSE: Several investigators have described intraoperative planning of prostate implants based on a nomogram. The aim of this work was to investigate the adequacy of the nomogram in predicting the total activity necessary for optimal dosimetry. METHODS AND MATERIALS: Eighty CT-based postimplant treatment plans were performed for patients who underwent ultrasound guided I-125 permanent implants alone between April 2000 and March 2001. The cohort of 40 patients had early stage (T1-T2) prostatic carcinoma and pre-treatment prostate volumes of 19 50 cc. I-125 seeds (0.391 mCi/seed) were implanted to achieve a distribution of 75% of the activity peripherally and 25% centrally. The CT studies were obtained on the day of (CT1) and at 1 month (CT2) after implant. All patients were catheterized at CT1, and 28 patients were catheterized at CT2 to visualize the urethra. For each patient, the percentage difference (dA) between the total implanted and nomogram predicted activity for a known prostate volume was calculated. The V200 (volume receiving 200% of the prescribed dose), V150, V100, V90, D100 (maximum dose received by 100% of the volume), D90, and D80 were measured for the prostate at CT1 and CT2. For the urethra, V275, V250, V200, and V150 were evaluated, and V100 and V70 were evaluated for the rectum. The Pearson test was used to correlate the dosimetric parameters with dA. Linear regression was used to fit the correlation of the volume and dose parameters with dA. RESULTS: The median V100 at CT1 and CT2 was 91.8% and 94.2%, respectively. The Pearson test was significant for the prostate V100 and dA measured at CT1 (p = 0.005) but not at CT2 (p = 0.106). A similar correlation was found for the prostate D90 at CT1 (p = 0.002), but not at CT2 (p = 0.076). D100 (maximum dose received by 100% of volume) for prostate did not correlate with dA at CT1 (p = 0.094) and CT2 (p = 0.148). The volume of the prostate receiving higher doses (greater than 150% and 200% of the prescribed dose) correlated with dA. There were no significant correlations between V275, V250, V200, and V150 at CT1 and CT2 as a function of dA for the urethra. V100 and V70 for the rectum correlated significantly with dA; for V100, p = 0.041 at CT1 and p = 0.014 at CT2 and for V70, p = 0.041 at CT1 and p = 0.026 at CT2. A linear regression model fitted to the prostate data obtained from CT1 with the goal of achieving a V100 of 90% and D90 of 145 Gy suggests that no increase in the number of seeds may be warranted using intraoperative planning. The implants examined showed no concomitant increase of urethral doses with increase in activity relative to the nomogram, but showed an increase in the rectal doses for the same increase in activity. CONCLUSION: The doses evaluated at CT1 represent an underestimate, whereas those obtained at CT2 represent an overestimate of the actual delivered protracted permanent implant dose. Based on these results and consideration of the dynamic nature of the dose distribution, target coverage obtained with intraoperative planning using the nomogram predicted activity is consistent with published guidelines for a quality implant and critical structure doses are within tolerance. PMID- 12738322 TI - Incomplete glandular ablation after salvage cryotherapy for recurrent prostate cancer after radiotherapy. AB - PURPOSE: To determine the clinical variables associated with inadequate salvage cryotherapy and to correlate the disease-free survival rates associated with biopsy results in prostate cancer (CaP). METHODS AND MATERIALS: Between July 1992 and January 1995, 150 patients underwent salvage cryotherapy for locally recurrent CaP. Biopsy specimens were examined for the presence of cancer cells and normal or atypical glands, all of which were considered evidence of inadequate cryotherapy. Clinical variables, as predictors of biopsy results, were evaluated with univariate and multivariate analyses. The impact of the biopsy results on disease-free survival was also determined. RESULTS: The number of cryoprobes and freeze-thaw cycles correlated with inadequate cryotherapy (p = 0.037 and p = 0.0022, respectively). The number of freeze-thaw cycles was an independent predictor of inadequate cryotherapy (p = 0.003). The finding of cancer cells in the biopsy specimens was the only histopathologic variable that affected disease-free survival (p = 0.016). CONCLUSION: Complete ablation of the prostate gland and tumor is difficult to achieve with salvage cryotherapy. To optimize for complete ablation, salvage cryotherapy should include at least two freeze-thaw cycles and a minimum of five cryoprobes. The finding of atypical or normal epithelial tissue in biopsy specimens after salvage cryotherapy is not predictive of biochemical failure. PMID- 12738323 TI - Significance of dose in postoperative radiotherapy for soft tissue sarcoma. AB - PURPOSE: For soft tissue sarcoma, adjuvant postoperative radiotherapy improves the local control rate over surgery alone. However, the issue of a dose-control relationship is controversial and was addressed in this study. METHODS AND MATERIALS: We retrospectively reviewed the records of 775 consecutive patients who received postoperative external beam radiotherapy and used univariate and multivariate analysis to determine whether a dose-control relationship exists for the whole group and for specific high-risk subgroups. RESULTS: With a median follow-up of 12.3 years, overall local control rates were 82%, 80%, and 79% at 5, 10, and 15 years. Factors adverse for local control were positive resection margins; tumor location in the head and neck and deep trunk; presentation with locally recurrent disease; patient age >64 years; histopathologic subtype of malignant fibrous histiocytoma, neurogenic sarcoma or epithelioid sarcoma; and tumor size >10 cm. In multivariate analysis, radiation dose > or = 64 Gy vs. <64 Gy independently correlated with improved local control. Significant interactions were noted between increased effectiveness of a higher dose and presentation with locally recurrent disease, with head and neck and deep trunk tumor sites, and with positive or uncertain resection margins such that local control was specifically improved under these circumstances with doses on the order of 64-68 Gy compared with doses on the order of 60 Gy. Higher dose, however, did not completely abrogate the adverse effect of positive margins. CONCLUSION: After gross total resection, soft tissue sarcoma with features predictive of a higher than average local recurrence rate benefited from doses of 64-68 Gy compared with 60 Gy. PMID- 12738324 TI - Preoperative vs. postoperative radiation therapy for soft tissue sarcoma: a retrospective comparative evaluation of disease outcome. AB - PURPOSE: Radiation (XRT) is a proven component in the treatment of soft tissue sarcoma. However, there is little evidence regarding the relative effectiveness of preoperative vs. postoperative XRT. This retrospective study addresses the relative effectiveness of disease control by these two treatment sequences. METHODS AND MATERIALS: A total of 517 patients (246 treated with postoperative XRT, and 271 treated with preoperative XRT) with nonmetastatic sarcoma were evaluated for disease outcome and late complications using univariate and multivariate techniques. RESULTS: With a median follow-up of 6 years, overall local control was 81% and 78% at 5 and 10 years, respectively. Although local control appeared superior with preoperative XRT (83% at 10 years) compared with postoperative XRT (72%), multivariate analysis revealed that this difference could be entirely explained by the unequal distribution of prognostic factors between the two groups, and there was no evidence that treatment sequence independently determined local control. There were no differences in nodal or metastatic relapse between the two treatments, and disease-specific survival was not significantly different. There was a slightly higher incidence of late XRT related complications among those treated with postoperative XRT (10-year incidence of 9% vs. 5%, p = 0.03). CONCLUSIONS: This study found no evidence for differences in disease outcome attributable to the use of either pre- or postoperative XRT. There was a slight increase in long-term complications with postoperative XRT, likely due to the higher doses used in this sequence. PMID- 12738325 TI - FDG-PET lymph node staging and survival of patients with FIGO stage IIIb cervical carcinoma. AB - PURPOSE: To evaluate the outcome of patients with International Federation of Gynecology and Obstetrics (FIGO) clinical Stage IIIb cervical carcinoma as a function of site of initial regional lymph node metastasis as detected by 2[18F]fluoro-2-deoxy-D-glucose (FDG)-positron emission tomography (PET). METHODS AND MATERIALS: Forty-seven patients with FIGO Stage IIIb cervical cancer were evaluated before therapy with whole-body FDG-PET. Most patients were treated with external beam irradiation, intracavitary brachytherapy, and weekly cisplatin for six cycles. Overall and cause-specific survival rates were calculated by the Kaplan-Meier method. RESULTS: The pretreatment whole-body FDG-PET demonstrated that all patients had FDG uptake in the cervix. Of 47 patients, 13 (28%) had no evidence of lymph node metastasis, 20 (43%) had metastasis to pelvic lymph nodes only, 7 (15%) had pelvic and para-aortic lymph node metastases, and 7 (15%) had metastases to pelvic, para-aortic, and supraclavicular lymph nodes. The 3-year estimate of cause-specific survival was 73% for those with no lymph node metastasis, 58% for those with only pelvic lymph node metastasis, 29% for those with pelvic and para-aortic lymph node metastases, and 0% for those with pelvic, para-aortic, and supraclavicular lymph node metastasis (p = 0.0005). CONCLUSION: The cause-specific survival for patients with FIGO Stage IIIb carcinoma is highly dependent on the extent of lymph node metastasis as demonstrated by whole-body FDG-PET. PMID- 12738326 TI - Significant correlation of hypoxia-inducible factor-1alpha with treatment outcome in cervical cancer treated with radical radiotherapy. AB - PURPOSE: In the early stages of cervical cancer treated with surgery alone, hypoxia-inducible factor-1alpha (hif-1alpha) expression is prognostic for overall survival. Because hypoxia plays an important role in radiation resistance, we investigated hif-1alpha expression in cervical cancer treated with local radical radiotherapy (RT). METHODS AND MATERIALS: Between 1990 and 1998, 91 patients with squamous cell or adenocarcinoma of the uterine cervix were treated with external beam RT with and without brachytherapy. Biopsies from 78 patients were available for immunohistochemistry. The impact of the immunoreactivity of hif-1alpha in regard to survival end points was determined by univariate and multivariate analyses. Correlations with clinicopathologic characteristics were determined by cross-tabulations. RESULTS: Hif-1alpha was expressed in 73 (94%) of 78 patients. It was closely linked to the pretreatment hemoglobin level (p = 0.04, r = -0.22, Spearman correlation test). The Kaplan-Meier curves showed a significantly shorter local progression-free survival (p = 0.04, log-rank) and overall survival (p = 0.01, log-rank) and a trend for shorter disease-free survival (p = 0.15) for patients with increased hif-1alpha expression. The multivariate analyses revealed hif-1alpha expression to be an independent factor for overall survival (p = 0.02). CONCLUSION: Hif-1alpha is expressed in the vast majority of patients with advanced cervical cancer and had a prognostic significance. A weak but significant correlation was noted with pretreatment hemoglobin level. PMID- 12738327 TI - Evaluation of the LENT-SOMA scales for the prospective assessment of treatment morbidity in cervical carcinoma. AB - PURPOSE: To examine the Late Effects Normal Tissue Task Force (LENT)-Subjective, Objective, Management, Analytic (SOMA) scales prospectively in carcinoma of the cervix treated curatively with radiotherapy (RT) using interviews and postal questionnaires and to test the sensitivity of the scales in assessing the radiation effects. METHODS AND MATERIALS: A consecutive series of 100 patients completed questionnaires to score the subjective part of the published LENT-SOMA scales. Assessments were made before RT and at approximately 21, 70, 200, 400, 600, and 800 days after the start of treatment. The acceptability and feasibility of using the scales was examined using compliance in completion of the questionnaires. The scales were validated by evaluating the concordance of data obtained by two independent scorers and by examining the ability of the scales to measure radiation-related symptoms. RESULTS: Questionnaires were completed for 89 patients before RT. The level of noncompliance was 11%. The concordance between scores when two people completed the questionnaires independently was excellent. Subjective subsite scores were highest 21 days after treatment but generally fell by 70 days. The average baseline overall LENT-SOMA subjective scores increased with advancing stage (p = 0.008) and were higher for patients treated with RT alone (p = 0.044). CONCLUSION: In cervical carcinoma, the LENT-SOMA scales were acceptable and feasible to administer in the clinic and appropriate in the measurement of early subjective morbidity from RT. PMID- 12738328 TI - Radiation therapy for intracranial germ cell tumors. AB - PURPOSE: To review the combined experiences of University of California, San Francisco, and Stanford University Medical Center in the treatment of intracranial germ cell tumors (GCT) and to assess the impact of craniospinal radiation (CSI) on patterns of relapse, progression-free survival (PFS), and overall survival (OS). PATIENTS AND METHODS: Ninety-three patients received radiation for newly diagnosed intracranial GCTs, including 49 germinomas, 16 nongerminomatous GCTs (NGGCT), and 28 with no biopsy. Median follow-up for surviving patients was 4.5 years (range 0.25-34). Tests for variables correlating with OS and PFS were conducted using Cox proportional hazards model. RESULTS: Five-year PFS and OS rates were 60% +/- 15% and 68% +/- 14% for patients with NGGCT and 88% +/- 5% and 93% +/- 4% for those with germinoma. Of 6 patients with localized NGGCT who did not receive CSI, 1 experienced an isolated spinal recurrence but was salvaged. Of 41 patients with localized germinoma, 6 who received CSI and 35 who did not, no isolated spinal cord relapses occurred. Twenty-one patients with localized germinoma received neither CSI nor whole brain radiation. Of these, none of 18 with ventricular radiation relapsed. One of 3 patients with primary tumor radiation relapsed intracranially but had only received 11 Gy at initial treatment. On multivariate analysis, germinoma histology but not CSI correlated with improved PFS and OS. CONCLUSION: CSI is not indicated in the treatment of localized germinomas. For patients with localized germinomas treated with radiation alone, we recommend ventricular irradiation followed by primary tumor boost to a total of 45-50 Gy. PMID- 12738329 TI - Hypofractionated radiotherapy for elderly or younger low-performance status glioblastoma patients: outcome and prognostic factors. AB - PURPOSE: To evaluate the outcome for elderly or younger poor prognosis glioblastoma patients treated with hypofractionated radiotherapy (HypoRT). METHODS AND MATERIALS: Retrospective review at The University of Texas M. D. Anderson Cancer Center identified 59 glioblastoma patients (median age 65 years) treated with HypoRT between 1988 and 2001 with 50 Gy given at 2.5 Gy/fraction/day in 20 fractions within 4 weeks. Classification was according to the Radiation Therapy Oncology Group recursive partitioning analysis and was Class IV in 11, V in 29, and VI in 19. Surgery consisted of gross total resection (n = 16), subtotal resection (n = 28), and biopsy only (n = 13). Two patients were treated presumptively on the basis of radiographic findings. Chemotherapy was given either as part of the initial treatment (n = 15) or for progression (n = 9). RESULTS: The median survival time for the entire study population was 7 months, and the median progression-free survival was 3.9 months. The median survival time for patients with Class IV, V, and VI was 11, 7, and 5 months, respectively. Concordance was found with Radiation Therapy Oncology Group-established recursive partitioning analysis class survival. Steroid requirements were not increased during RT compared with preoperatively and immediately postoperatively. Late complications of HypoRT were limited to 3 cases of radiation necrosis suggested by MRI, 2 of which were pathologically confirmed. CONCLUSION: HypoRT consisting of 50 Gy in 4 weeks can be used for selected GBM patients to reduce the overall treatment time of conventional RT by 33-39% without apparent increased toxicity or decrement in survival. PMID- 12738330 TI - Repeat linear accelerator radiosurgery for cerebral arteriovenous malformations. AB - PURPOSE: To evaluate repeat radiosurgery (RS2) for cerebral arteriovenous malformations (AVMs) after failure of initial radiosurgery (RS1). METHODS AND MATERIALS: Between 1986 and 2000, 41 patients underwent RS2. Nine patients were not assessable: seven had insufficient follow-up (RS2 in 1999 and 2000) and two had no recent control angiography data. Thus, 32 (78%) of 41 patients were assessed. Most lesions (29 [90%] of 32) were supratentorial: 22 (69%) on the left, 8 (25%) on the right, and 2 on the midline (6%). The patients had Spetzler Martin Grade 1-5 (median Grade 3). The symptoms before RS1 included hemorrhage in 20 (63%), epilepsy in 10 (31%), progressive neurologic deficits in 2 (6%), and headaches in 6 (19%). Five patients had two or more symptoms. Twenty-two patients (69%) had received other treatment before RS1, including neurosurgery in 3 patients (9%) and one to six embolizations in 19 patients. At RS1, the median largest nidus dimension was 2.7 cm (range 0.8-5). The median volume was 2.7 cm(3) (range 1.2-9.9). The median time from RS1 to RS2 was 52 months (range 12-126). Between RS1 and RS2, 7 (22%) of 32 patients experienced bleeding. The same irradiation technique was used for RS1 and RS2, except for 2 patients who underwent RS2 at another institution. Circular 15-MV X-ray minibeams (range 6-20 mm) and coronal arcs were used. RS1 was monoisocentric in 75% of cases and multiisocentric in 25%. At RS2, the median largest nidus dimension was 3 cm (range 1.4-5). The median volume was 4.2 cm(3) (range 0.8-13.4). RS2 was monocentric in 72% of cases and multiisocentric in 28%. After RS2, the median follow-up was 19.5 months (range 0-79; mean 25.3). RESULTS: After RS2, the obliteration rate was 59.3% (19 of 32). The median time to arteriographic obliteration was 21 months (range 12-96). The survival rate was 97% (31 of 32). Five of the 13 patients with a nonobliterated nidus experienced complications; 3 had bleeding (9%) and 2 without prior neurologic deficits developed partially regressive neurologic deficits. One patient with a previously existing deficit developed an additional new partially regressive neurologic deficit after an episode of bleeding. Thus, 3 (9%) of 32 patients had neurologic complications. Moderate-grade parenchymal changes at MRI increased after RS2 (88.2% vs. 57.7% after RS1; p = 0.10, not significant). However, necrosis-like changes did not significantly increase. After RS1 failure, salvage may be attempted by embolization, neurosurgery, or RS2. RS2 should be considered after the second successive annual angiogram if reduction of the nidus is <25%. CONCLUSION: The results after RS2 are encouraging. A multidisciplinary approach is mandatory to reduce the initial failure rate and to choose the modality and timing of salvage treatment. PMID- 12738331 TI - Preliminary visual outcomes after three-dimensional conformal radiation therapy for optic nerve sheath meningioma. AB - PURPOSE: We assessed visual outcomes, local control, and toxicity associated with three-dimensional conformal radiation therapy (3D-CRT) for primary optic nerve sheath meningiomas (ONSM). METHODS: Twenty-three patients diagnosed with ONSM were evaluated at the University of Michigan between 1986 and 2001. Fourteen patients were treated with 3D-CRT. Detailed pre- and postradiation treatment ophthalmologic examinations and MRIs were performed on all patients. Clinically significant visual acuity change was defined as a >or=three line change on the Snellen chart. Mean deviation change of >or=three decibels was defined as a clinically significant visual field change. Radiographic progression was defined as any increase in size on MRI. Acute and late toxicity was scored according to RTOG criteria. RESULTS: Median follow-up was 51.3 months. Five patients had a clinically significant improvement in visual acuity. Seven had stable acuity, and only 2 worsened. Nine patients had clinically significant visual field improvement. One patient developed early radiation retinopathy, 1 experienced orbital pain, 1 developed dry eye, and 2 developed iritis. No patient has required additional treatment, and none have demonstrated radiographic progression. CONCLUSION: 3D-CRT is effective in controlling tumor growth while improving or preserving vision in most patients with optic nerve sheath meningiomas. PMID- 12738332 TI - The American Brachytherapy Society recommendations for brachytherapy of uveal melanomas. AB - PURPOSE: This article presents the American Brachytherapy Society (ABS) guidelines for the use of brachytherapy for patients with choroidal melanomas. METHODS: Members of the ABS with expertise in choroidal melanoma formulated brachytherapy guidelines based upon their clinical experience and a review of the literature. The Board of Directors of the ABS approved the final report. RESULTS: Episcleral plaque brachytherapy is a complex procedure and should only be undertaken in specialized medical centers with expertise in this sophisticated treatment program. Recommendations were made for patient selection, techniques, dose rates, and dosages. Most patients with very small uveal melanomas (<2.5 mm height and <10 mm in largest basal dimension) should be observed for tumor growth before treatment. Patients with a clinical diagnosis of medium-sized choroidal melanoma (between 2.5 and 10 mm in height and <16 mm basal diameter) are candidates for episcleral plaques if the patient is otherwise healthy and without metastatic disease. A histopathologic verification is not required. Small melanomas may be candidates if there is documented growth; some patients with large melanomas (>10 mm height or >16 mm basal diameter) may also be candidates. Patients with large tumors or with tumors at peripapillary and macular locations have a poorer visual outcome and lower local control that must be taken into account in the patient decision-making process. Patients with gross extrascleral extension, ring melanoma, and tumor involvement of more than half of the ciliary body are not suitable for plaque therapy. For plaque fabrication, the ophthalmologist must provide the tumor size (including basal diameters and tumor height) and a detailed fundus diagram. The ABS recommends a minimum tumor (125)I dose of 85 Gy at a dose rate of 0.60-1.05 Gy/h using AAPM TG-43 formalism for the calculation of dose. NRC or state licensing guidelines regarding procedures for handling of radioisotopes must be followed. CONCLUSIONS: Brachytherapy represents an effective means of treating patients with choroidal melanomas. Guidelines are established for the use of brachytherapy in the treatment of choroidal melanomas. Practitioners and cooperative groups are encouraged to use these guidelines to formulate their treatment and dose reporting policies. These guidelines will be modified as further clinical results become available. PMID- 12738333 TI - Hodgkin's disease in elderly patients (> or =60): clinical outcome and treatment strategies. AB - PURPOSE: Older age is an adverse prognostic factor for survival for patients with Hodgkin's disease. This study assessed the outcome of elderly patients (>or=60 years) with Hodgkin's disease treated with curative intent in an attempt to identify the optimal treatment strategies for this group of patients. METHODS AND MATERIALS: Eighty-six patients, 60-93 years old at the time of diagnosis, were treated for Hodgkin's disease with radical intent between 1969 and 1995. All patients underwent radiographic staging of the chest, abdomen, and pelvis, and 17 patients underwent staging laparotomy. Fifty-two patients had early-stage disease (Stage IA-IIA) and 34 had Stage IIB-IV Hodgkin's disease. The median follow-up time was 75 months (range 24-267) for surviving patients. RESULTS: The 10-year actuarial freedom from treatment failure (FFTF) rate for all patients was 62%. The 10-year FFTF rate for patients with Stage IA-IIA and Stage IIB-IV disease was 71% and 49%, respectively (p = 0.03). Patients with early-stage disease treated with chemoradiotherapy had a lower crude rate of treatment failure (20%) than patients treated with either chemotherapy alone (33%) or radiotherapy alone (46%). However, no statistically significant difference was found between the treatment groups in terms of actuarial FFTF or overall survival. The 5- and 10 year overall survival rate (all causes) for all patients was 48% and 30%, respectively. The 10-year survival rate for patients with Stage IA-IIA and Stage IIB-IV disease was 31% and 26%, respectively (p = 0.07). On multiple regression analysis, including age, treatment, and stage in the Cox regression model with respect to overall survival, age was a marginally significant factor (p = 0.08). For FFTF, age was not a significant factor in the model (p = 0.11). We analyzed the subsequent outcome of patients who developed a first recurrence after initial treatment; the 5-year survival rate was only 20% after recurrence of Hodgkin's disease. Initial treatment was reasonably well tolerated. CONCLUSION: Although more patients died of other causes than Hodgkin's disease, the recurrence of Hodgkin's disease had a significant impact on survival. Thus, we favor the use of chemoradiotherapy in early-stage patients >60 years to minimize the risk of relapse. PMID- 12738334 TI - Fibrogenic signals in patients with radiation enteritis are associated with increased connective tissue growth factor expression. AB - PURPOSE: To investigate the expression of a new fibrogenic cytokine the connective tissue growth factor (CTGF) in intestinal radiation fibrosis and to characterize the mesenchymal cell subtypes involved in CTGF synthesis and collagen deposition. METHODS AND MATERIALS: Sixteen patients with radiation enteritis that occurred after radiotherapy for pelvic malignancies and 6 with histologically normal bowel entered the study. Immunohistochemistry, Western blot analysis, and real-time reverse transcriptase-polymerase chain reaction were performed to study CTGF expression, along with other known markers of radiation fibrosis: the pro-fibrogenic cytokine transforming growth factor (TGF)-beta1 and phenotypic markers of the fibroblast differentiation the alpha-sm actin (A), vimentin (V), and desmin (D). Finally, the collagen accumulation was measured by Sirius red staining and colorimetric assay. RESULTS: Radiation enteritis was characterized by increased collagen content within the intestinal wall. CTGF immunoreactivity, protein, and mRNA level were increased in radiation enteritis compared with the healthy bowel. On the contrary, no increase of the TGF-beta1 mRNA level was observed in radiation enteritis compared with healthy bowel, and the level of TGF-beta protein was slightly increased in radiation enteritis. A co localization of CTGF immunoreactivity and collagen deposition was found in the extracellular matrix and subtypes of activated mesenchymal cells with a fibroblast phenotype (V(+)/D(-)/A(-)) and myofibroblast phenotype (V(+)/D( /+)/A(+)). CONCLUSION: The increased level of CTGF protein and mRNA associated with the accumulation of fibroblasts/myofibroblasts and collagen deposition were parts of the fibrogenic signals involved in the persistence of late intestinal radiation fibrosis. PMID- 12738336 TI - Analysis of interaction between number of implant catheters and dose-volume histograms in prostate high- dose-rate brachytherapy using a computer model. AB - PURPOSE: In prostate high-dose-rate brachytherapy, to determine before implant, using the standard geometric optimization algorithm, whether there is an optimal number of catheters. MATERIALS AND METHODS: Transrectal ultrasound images of the prostate from 24 patients were transferred into the brachytherapy planning system. Urethra and prostate contours were digitized onto each axial slice of a CT scan, as well as hypothetical locations of the catheters (2/3 of the catheters along the prostate contour, 1/3 around the urethra). Each prostate was implanted with 9, 12, 15, 18, and 21 catheters. Dosimetry was optimized using a geometric optimization algorithm; prescription isodose was chosen so that 95% of planning target volume was covered by the 100% isodose. RESULTS: A significant increase in mean volume of prostate receiving 150% of the dose (V150) when the number of catheters decreased (p < 0.0001). The 9-catheter group significantly differed from each of the other groups; no difference was seen in V150 among the 21-, 18-, and 15-catheter groups. Parallel results were observed for urethra V150 and homogeneity index; there was no difference in conformity index by catheter group. CONCLUSION: V150 increased when fewer catheters were used. There was no significant difference among the 21-, 18-, and 15-catheter groups: the geometric optimization routine probably compensated for the larger distance between dwell positions. Based on the technique described in our study, we conclude that 15 to 21 catheters seem to cover the prostate adequately without creating excess hot spots. PMID- 12738335 TI - Simultaneous integrated boost intensity-modulated radiotherapy for locally advanced head-and-neck squamous cell carcinomas. I: dosimetric results. AB - PURPOSE: This report describes the dosimetric analyses of a Phase I/II protocol, designed to examine the capabilities of an institutionally developed intensity modulated radiotherapy (IMRT) system with respect to dose escalation. The protocol employed stringent dosimetric guidelines in the treatment of locally advanced head-and-neck squamous cell carcinomas (HNSCC) with radiotherapy alone using IMRT and the simultaneous integrated boost (SIB) technique. METHODS AND MATERIALS: The first 14 patients enrolled on the protocol were included in this analysis. Escalating doses of 68.1 Gy (6 patients), 70.8 Gy (6 patients), and 73.8 Gy (2 patients) were delivered to the gross tumor volume (GTV) in 30 fractions. Simultaneously, constant dose coverage was given to the subclinical disease and the electively treated nodal regions, which received 60 Gy and 54 Gy, respectively, in all three cohorts. Parotid glands were spared to the degree possible without compromising target coverage. The following indices are reported for the GTV: (1) dose to specified percent volumes (e.g., D(98) and D(2)); (2) homogeneity index defined as the ratio (D(2) - D(98))/D(prescription); (3) biologically equivalent uniform dose (EUD); and (4) an index of conformality, PITV, defined as the ratio of volume enclosed within the prescribed isodose surface to the target volume. Treatments were planned and delivered with nine 6 MV photon beams using the multileaf collimator (MLC) "sliding window" technique. RESULTS: Mean doses to 98% of GTV were 68.4 Gy, 70.5 Gy, and 70.8 Gy, and average GTV dose homogeneity was 6.7%, 7.6%, and 8.8% for the three cohorts. The average doses to the parotid gland proximal to and distant from GTV were 41.3 Gy and 25.7 Gy, respectively. Dose distributions measured in phantom showed good agreement with calculations. CONCLUSIONS: Treatment of locally advanced HNSCC using SIB IMRT as described is feasible. Treatment planning and delivery are safer and more efficient than with conventional three-dimensional processes. Predicted dose distributions can be accurately delivered with excellent conformality using dynamic MLC. At least one of the parotid glands can be adequately spared. Patient follow-up continues and will allow eventual quantitative correlation of delivered dose distributions with clinical outcomes. PMID- 12738337 TI - Commissioning and clinical results utilizing the Gildenberg-Laitinen Adapter Device for X-ray in fractionated stereotactic radiotherapy. AB - BACKGROUND: The Gildenberg-Laitinen Adapter Device for X-Ray (GLAD-X/LS) frame is a positioning device that allows the use of the same fiducial points as the Brown Robert-Wells (BRW) system. Thus it permits treatment planning to be accomplished by the Radionics X-knife Radiosurgery Program. We investigated the commissioning and clinical benefits of the GLAD-X/LS for fractionated stereotactic radiotherapy (FSRT) in patients who were unable to tolerate the Gill-Thomas-Cosman (GTC) frame. METHODS AND MATERIALS: Commissioning of the GLAD-X/LS system was done via use of a Rando Phantom. A target volume of 2 x 2 x 2 cm was drilled into the phantom head. An ion chamber and thermoluminescence dosimetric chips (TLDs) were implanted in the target. A simulated treatment course consisting of 5 stereotactic radiotherapy fractions (300 cGy, 30 mm collimator) was delivered to the phantom head. A total of 27 patients who could not tolerate the GTC frame were treated using the GLAD-X/LS system. A total of 35 isocenters were used; the median number of treatment fractions was eight. Reproducibility of the x, y, and z coordinates was examined and correlated to the same determined using orthogonal port films. Relocation accuracy and reproducibility were further assessed comparing the x, y, and z coordinates of the target center with multiplanar reconstructed coronal and sagittal images. Patient tolerance of the device was also evaluated daily throughout the treatment. RESULTS: The measured TLD and ion chamber doses were within 3% of the prescribed dose at the isocenter. The same dose accuracy was also found at incremental distances of 5 mm, 10 mm, and 15 mm from the isocenter. All patients tolerated the treatment and the device well. Six patients experienced mild ear canal pain, and softer or smaller earpieces were substituted. The mean relocation accuracy was 1.5 mm +/- 0.8. CONCLUSIONS: The GLAD-X/LS system has excellent accuracy and reproducibility with the mean relocation accuracy of 1.5 mm +/- 0.8. The device is well-tolerated by patients, with no significant complications. Larger scale studies are necessary before routine use can be recommended for the administration of FSRT. PMID- 12738338 TI - Evaluating radiation therapy in prostate cancer. PMID- 12738340 TI - Brain activation patterns during a selective attention test-a functional MRI study in healthy volunteers and patients with schizophrenia. AB - Functional magnetic resonance imaging was used to compare cortical activation patterns in healthy volunteers with those in patients with schizophrenia during a modified verbal Stroop task. Healthy subjects (n=13) and patients with schizophrenia (n=13) on stable antipsychotic treatment, matched on demographic variables, were included. Patients were preselected on the basis of good performance on a selective attention test. Patients with schizophrenia showed a significantly increased pattern of activation in the left and right inferior frontal cortex and the anterior cingulate cortex. A significant negative correlation between activation of the left prefrontal cortex and accuracy in the modified Stroop test was observed for healthy controls but not schizophrenia patients. Although both groups recruited the prefrontal cortex during the modified Stroop task, for the schizophrenia patients this activation was bilateral, whereas for the controls this activation was primarily in the left hemisphere, suggesting that patients with schizophrenia recruited more prefrontal regions to perform the task with the same accuracy as healthy controls. Our findings of increased activity across multiple areas of the brain, including dorsolateral frontal cortex and anterior cingulate, in patients with schizophrenia who perform relatively well on a task of selective attention give further evidence that task performance may be a confounding factor in the interpretation of neuroimaging results. PMID- 12738341 TI - Why are smiles contagious? An fMRI study of the interaction between perception of facial affect and facial movements. AB - In human communication there is often a close relationship between the perception of an emotionally expressive face and the facial response of the viewer himself. Whereas perception and generation of facial expressions have been studied separately with functional imaging methods, no studies exist on their interaction. We combined the presentation of emotionally expressive faces with the instruction to react with facial movements predetermined and assigned. fMRI was used in an event related design to examine healthy subjects while they regarded happy, sad, or neutral faces and were instructed to simultaneously move the corners of their mouths either (a). upwards or (b). downwards, or (c). to refrain from movement. The subjects' facial movements were recorded with an MR compatible video camera. Movement latencies were shortened in congruent situations (e.g. the presentation of a happy face and combined with upward movements) and delayed in non-congruent situations. Dissonant more than congruent stimuli activated the inferior prefrontal cortex and the somatomotor cortex bilaterally. The congruent condition, in particular when seeing a happy face, activated the medial basotemporal lobes (hippocampus, amygdala, parahippocampal region). We hypothesize that this region facilitates congruent facial movements when an emotionally expressive face is perceived and that it is part of a system for non-volitional emotional facial movements. PMID- 12738342 TI - Association between regional cerebral blood flow and eye-tracking performance and the Wisconsin Card Sorting Test in schizophrenics: a single photon emission computed tomography study. AB - The objectives of this study were (1). to examine the changes in regional cerebral blood flow (rCBF) during Wisconsin Card Sorting Test (WCST) performance in two different eye-tracking groups; (2). to explore the relationship between eye-tracking movement and rCBF at rest; and (3). to estimate the association between WCST performance and rCBF in patients with schizophrenia. A total of 17 patients with schizophrenia were recruited. SPECT with Tc-99m HMPAO (Tc-99m hexamethylpropyleneamine oxime) was carried out while patients were performing the WCST and resting. Brodmann area 9 of the prefrontal cortex, a part of the dorsal lateral prefrontal cortex (DLPFC), was less activated during performance of the WCST in poor trackers (relative to good trackers). The eye pursuit tracking error measure in schizophrenic patients was negatively associated with decreases in rCBF in the middle temporal area, superior parietal lobule, thalami, and caudate nuclei. The rCBF increased significantly in the superior temporal gyri, inferior parietal lobe, and some frontal regions during WCST performance; however, this was not the case in the DLPFC. Additionally, significant correlations were found between WCST scores and rCBF during WCST performance in the prefrontal lobes, and in thalamic and cerebellar regions. Our findings suggest that the rCBF changes during WCST performance may be distinctive in different eye-tracking groups. Our results confirm the hypothesis that the middle temporal area, superior parietal lobule, thalami, and caudate nuclei-mainly parts of the oculomotor circuit-are involved in eye pursuit tracking. Surprisingly, no significant association was found in the frontal eye field. Although the frontal lobe plays a significant role in WCST performance, our findings demonstrate that WCST performance is widely involved with other regions in patients with schizophrenia. PMID- 12738343 TI - The Danish PET/depression project: poor verbal fluency performance despite normal prefrontal activation in patients with major depression. AB - The verbal fluency (VF) test is associated with prefrontal and cingulate functioning. We used positron emission tomography (PET) to test the hypothesis that inactivity in these regions can explain why patients with depression often perform poorly on this test. Forty-one patients with major depression and 46 controls were scanned during rest and during activation with the VF test. The differences between the two conditions were mapped for each of the two groups, and between-group differences in the activation pattern were calculated on a voxel-by-voxel basis. As predicted, the patients performed significantly more poorly on the test. In both groups activations were seen in the left anterior cingulate region, the left dorsolateral prefrontal cortex, the left medial prefrontal cortex, and the right cerebellum. Performance was correlated to the cerebral perfusion in to the left occipitotemporal gyrus and the left cerebellum. However, no difference in activation between the two groups was significant. The present study had sufficient power to detect potential differences between the two groups, and the subtraction tasks were appropriate. Therefore, the result supports the notion that abnormalities in the prefrontal cortex in depression are qualitative in nature rather than quantitative; this suggests depression involves dys-coordination of neural activity in the frontal lobes rather than a simple reduction in activity. PMID- 12738344 TI - Rightward cerebral asymmetry in subtypes of schizophrenia according to Leonhard's classification and to DSM-IV: a structural MRI study. AB - Although well documented, brain structural abnormalities in schizophrenia are non specific, and morphometric parameters show significant overlap between patients and healthy controls. Such inconsistencies in neuroimaging findings could represent different levels of severity along a single pathogenic process or distinct clinical and etiopathological psychoses within a schizophrenic spectrum. The aim of the present study was the investigation of distinct brain abnormalities in different subtypes of schizophrenia. Forty patients were classified according to DSM-IV and Leonhard's classifications. Psychopathology was assessed by the Positive and Negative Syndrome Scale (PANSS) and the Negative Symptom Rating Scale (NSRS). Patients were compared to 20 healthy volunteers on volumetric measures of cerebral structures (hemisphere, hippocampus and planum temporale) and ventricular-brain ratio (VBR) obtained by magnetic resonance imaging. Patients showed rightward asymmetry of cerebral hemispheres and increased VBR. Rightward asymmetry correlated with severity of negative symptoms and prevailed in the systematic forms of Leonhard, suggesting a distinct pattern of left hemisphere abnormality in this subgroup of psychoses. Increased VBR values showed a single normal distribution in the subgroups, indicating that ventricular enlargement is not restricted to a subgroup but is present to a certain degree in all cases. PMID- 12738345 TI - Foreword to: buprenorphine and buprenorphine/naloxone: a guide for clinicians. PMID- 12738347 TI - The clinical pharmacology of buprenorphine: extrapolating from the laboratory to the clinic. AB - This paper will review clinical pharmacology studies on buprenorphine, a mixed opioid agonist-antagonist currently approved as a treatment for opioid dependence. The focus is on studies characterizing buprenorphine's pharmacodynamic actions, including its safety, abuse liability, withdrawal suppression and withdrawal precipitation capacity, physical dependence potential, cross-tolerance and duration of action as well as a review of the pharmacological profile of buprenorphine/naloxone combinations. The findings from these clinical pharmacology studies are synthesized and presented in a framework designed to (1) inform clinicians about the advantages and disadvantages of buprenorphine as an opioid maintenance agent, and (2) provide information about dosing procedures that may optimize the use of buprenorphine in the clinic. PMID- 12738346 TI - From morphine clinics to buprenorphine: regulating opioid agonist treatment of addiction in the United States. AB - The practice of prescribing opioid drugs for opioid dependent patients in the U.S. has been subjected to special government scrutiny for almost 100 years. From 1920 until 1964, doctors who used opioids to treat addicts risked federal and/or state criminal prosecution. Although that period ended when oral methadone maintenance was established as legitimate medical practice, public concern about methadone diversion and accidental overdose fatalities, combined with political pressure from both hostile bureaucracies and groups committed to drug-free treatments, led to the development of unprecedented and detailed Food and Drug Administration (FDA) regulations that specified the manner in which methadone (and later, levo-alpha-acetyl methadol, or levomethadyl acetate, (LAAM)) could be provided. In 1974, Congress gave the Drug Enforcement Administration (DEA) additional oversight of methadone treatment programs. Efforts to liberalize the FDA regulations over the past 30 years have been resisted by both the DEA and existing treatment providers. Additional flexibility for clinicians may evolve from the most recent effort to create an accreditation system to replace some of the FDA regulations. The development of buprenorphine, a partial opioid agonist, as an effective treatment for opioid addiction reopened the possibility for having a less burdensome oversight process, especially because of its reduced toxicity if ingested by non-tolerant individuals. New legislation, the Drug Addiction Treatment Act (DATA) of 2000, created an opportunity for clinicians with special training to be exempted from both federal methadone regulations and the requirement to obtain a special DEA license when using buprenorphine to treat addicts. Some details of how the DATA was developed, moved through Congress, and signed into law are described. PMID- 12738348 TI - Clinical and pharmacological evaluation of buprenorphine and naloxone combinations: why the 4:1 ratio for treatment? AB - Although only a partial mu-opiate agonist, buprenorphine can be abused and diverted from medical therapy to the illicit drug market. A combination of buprenorphine and naloxone for sublingual administration may discourage diversion and abuse by precipitating opiate withdrawal when taken parenterally. Because opiate-abusing populations are not homogeneous and have varying levels of opiate dependence, the efficacy of buprenorphine and naloxone in precipitating opiate withdrawal or in attenuating the pleasurable effects of buprenorphine may vary. This chapter describes the effects of sublingual and parenteral buprenorphine and naloxone combinations in several populations of opiate-dependent people. We conclude that buprenorphine and naloxone combinations should not diminish the efficacy of sublingual buprenorphine, but should have lower abuse liability than buprenorphine alone. PMID- 12738349 TI - Pharmacokinetics of the combination tablet of buprenorphine and naloxone. AB - The sublingual combination tablet formulation of buprenorphine and naloxone at a fixed dose ratio of 4:1 has been shown to be as effective as the tablet formulation containing only buprenorphine in treating opiate addiction. The addition of naloxone does not affect the efficacy of buprenorphine for two reasons: (1) naloxone is poorly absorbed sublingually relative to buprenorphine and (2) the half-life for buprenorphine is much longer than for naloxone (32 vs. 1 h for naloxone). The sublingual absorption of buprenorphine is rapid and the peak plasma concentration occurs 1 h after dosing. The plasma levels for naloxone are much lower and decline much more rapidly than those for buprenorphine. Increasing dose results in increasing plasma levels of buprenorphine, although this increase is not directly dose-proportional. There is a large inter-subject variability in plasma buprenorphine levels. Due to the large individual variability in opiate dependence level and the large variability in the pharmacokinetics (PK) of buprenorphine, the effective dose or effective plasma concentration is also quite variable. Doses must be titrated to a clinically effective level for individual patients. PMID- 12738350 TI - Clinical efficacy of buprenorphine: comparisons to methadone and placebo. AB - Buprenorphine has been studied extensively since 1978 when it was initially proposed as an alternative to methadone for treatment of opioid dependence. Early work by Jasinski et al., 1978; Mello and Mendelson, 1980; Mello et al., 1982; Mello et al., 1983 and Mendelson et al., 1984 and their colleagues demonstrated buprenorphine's low physical abuse potential and its ability to substitute for heroin and reduce heroin self-administration in opiate-dependent humans. The subsequent early clinical studies suggested that, in clinical settings, buprenorphine was a safe and efficacious opiate dependence pharmacotherapy. Formal approval for general clinical use, however, required that systematic data be gathered on buprenorphine's safety and efficacy in larger groups and a series of controlled clinical trials was designed to evaluate its utility from a medication development perspective. In general, these trials adhered to one of three basic protocol designs: comparison of buprenorphine to methadone; dose comparisons using dose response as an indicator of efficacy; and comparison of buprenorphine to placebo. Retention in treatment, reduction in illicit drug use and craving, and patient and staff ratings of improvements were the most frequently used outcome indicators in these trials. Additional data collected included optimum dosing and dosage schedules, adverse reactions and common side effects, and other information intended to clarify buprenorphine's benefit-risk relationship and to help prepare guidelines for its safe marketing and utilization by physicians in general clinical practice. This paper presents a review of the buprenorphine/methadone comparison trials conducted in the United States and two such trials conducted in Europe. Also reviewed are three placebo controlled trials and a buprenorphine/methadone detoxification study. Overall, this series of studies did firmly establish the efficacy of buprenorphine alone and in comparison to methadone. PMID- 12738351 TI - Buprenorphine: how to use it right. AB - The unique pharmacology of buprenorphine at the mu-opioid receptor (i.e. high affinity, low intrinsic activity and slow dissociation) results in buprenorphine having: (1) a good safety profile, (2) low physical dependence, and (3) flexibility in dose scheduling. Early studies assessed the effectiveness of buprenorphine for the treatment of opioid dependence using a sublingual solution formulation. More recently, a combination tablet (buprenorphine/naloxone in a 4:1 ratio) has been assessed with the goal of decreasing diversion and abuse. Controlled studies with buprenorphine solution, buprenorphine mono-tablet, and buprenorphine/naloxone combination tablet have uniformly demonstrated the effectiveness of buprenorphine for opioid dependence treatment and the combination tablet appears to decrease (but not eliminate) abuse potential. There is general agreement across studies regarding buprenorphine induction and maintenance dose schedules. The clinical effects of buprenorphine and buprenorphine/naloxone are similar and most patients can be treated initially with and maintained on a daily buprenorphine/naloxone dose of 4:1-24:6 mg. Dosing is possible on a less-than-daily schedule; however, multiples of the daily-dose should be administered to cover the increased interval between doses. If buprenorphine withdrawal is indicated, gradual dose reduction is recommended over a rapid dose reduction or abrupt cessation. Both tablet formulations are approved by the US FDA for opioid dependence treatment as Schedule III narcotics and are, therefore, available for use in office-based practice. The buprenorphine plus naloxone combination product should provide additional safeguards for use in office-based practice by decreasing risk of diversion, and office-based treatment should expand the availability of services to opioid dependent patients. PMID- 12738352 TI - Safety and health policy considerations related to the use of buprenorphine/naloxone as an office-based treatment for opiate dependence. AB - Opiate dependence remains a fundamental challenge confronting health delivery systems and is often characterized as a social and moral issue. The impact of this disorder on healthcare policy is changing with the increased incidence of HIV, hepatitis C, and tuberculosis infections in opiate-dependent patients. These medical illnesses have substantial effect on escalating healthcare costs, and, therefore, also affect healthcare policy priorities, which are responsive to these costs. Pharmacological treatments for opiate dependence have had limited success; often the consequence of limited access to care. Hence, there is a need to develop new pharmacotherapies for opiate dependence that extend the range of clinical options, including new first-line treatment approaches. This paper will focus on the safety and health policy considerations related to the use of buprenorphine and buprenorphine/naloxone based on data derived from clinical trials and post-marketing surveillance that provide evidence for the use of the medications as first-line treatments in an office-based environment. The evaluation of this evidence formed the basis by the National Institute on Drug Abuse to support and pursue the evaluation and registration of buprenorphine/naloxone and buprenorphine in a public/private sector cooperative effort to become an office-based, first-line treatment for opiate dependence. PMID- 12738353 TI - Use of buprenorphine in pregnancy: patient management and effects on the neonate. AB - It is estimated that 55-94% of infants born to opioid-dependent mothers in US will show signs of opioid withdrawal. Buprenorphine has been reported to produce little or no autonomic signs or symptoms of opioid withdrawal following abrupt termination in adults. To date, there have been 21 published reports representing approximately 15 evaluable cohorts of infants exposed to buprenorphine in utero. Of approximately 309 infants exposed, a neonatal abstinence syndrome (NAS) has been reported in 62% infants with 48% requiring treatment; apparently greater than 40% of these cases are confounded by illicit drug use. The NAS associated with buprenorphine generally appears within 12-48 h, peaks at approximately 72-96 h, and lasts for 120-168 h. These results appear similar to or less than that observed following in utero exposure to methadone. From a review of the literature, buprenorphine appears to be safe and effective in both mother and infant with an NAS that may differ from methadone both qualitatively and quantitatively. PMID- 12738354 TI - Accessing opiate dependence treatment medications: buprenorphine products in an office-based setting. PMID- 12738355 TI - Multiparameter precursor analysis of T-cell responses to antigen. AB - Triggering of the T-cell receptor by cognate antigen induces a variety of cellular events leading to cell proliferation and differentiation. While the plasticity and diversity of T-cell responses have been recognized for a long time, few quantitative studies have been conducted to measure what proportion of specific T cells will enter a given differentiation program after antigen stimulation. In the present study, we analyzed human T cells cultured with influenza-peptide-loaded dendritic cells. We compared three individual methods for assaying the frequency of antigen-specific T cells: ELISPOT, tetramer binding, and proliferation. The three methods yielded similar but not identical results. In order to study these differences at the single cell level, we developed a multiparameter flow cytometric method, which allows simultaneous analysis of antigen-specific tetramer binding, T-cell proliferation, and cytokine production. Based on these data, we used flow precursor frequency analysis to calculate the proportion of eight different precursor subsets in the original, resting population. We conclude that approximately half of the cells that bound specific tetramers actually proliferated and synthesized IFNgamma in response to antigen. In addition, similar numbers of cells that did not bind tetramer proliferated (but did not synthesize IFNgamma). The method allows for an estimate of the precursor frequency of each functional subset within the initial population. It could be applied to additional markers of function and differentiation, combining all parameters into a description of the complex response potential of a T-cell pool. PMID- 12738357 TI - Estimation of the relative affinity of B cell receptor by flow cytometry. AB - We have developed a simple method using flow cytometry to estimate the relative affinity of B cell receptor (BCR) possessing the hapten-binding activity. Bovine serum albumin (BSA) was conjugated with a hapten, (4-hydroxy-3-nitrophenyl)acetyl (NP) and biotin (NP-BSA-bio). The interaction between NP-BSA-bio and anti-NP monoclonal antibodies (mAb) was studied as a model of the BCR reaction by surface plasmon resonance (SPR) using a biosensor chip immobilized with mAbs through anti Fc antibody (Ab). The relative affinity of these mAbs was estimated on the basis of resonance units for the binding of NP(0.5)-BSA-bio(21) relative to that of NP(7.4)-BSA-bio(21) expressed as a ratio (NP(0.5)-BSA-bio(21)/NP(7.4)-BSA bio(21)). In combination with streptavidin (SA)-R-phycoerythrin (PE), we measured the binding of NP-BSA-bio to BCR by flow cytometry and found that a high number of biotin molecules was necessary to improve the sensitivity of detection of the bound NP-BSA-bio without steric hindrance in the NP-BCR interaction. We demonstrated that the ratio of the mean fluorescence intensity (MFI) of NP(0.5) BSA-bio(21)/NP(7.4)-BSA-bio(21) at a concentration of 10(-8) M could be used as a practical measure of the affinity. This method is expected to be useful for the study of affinity maturation on the cellular level. PMID- 12738356 TI - Comparison of analytical methods for the evaluation of antibody responses against epitopes of polymorphic protein antigens. AB - Surface exposed protein antigens of the malaria parasite Plasmodium falciparum frequently harbor multiple dimorphic amino acid positions. These are associated with parasite immune evasion and represent a major obstacle for subunit vaccine design. Here, we have analyzed the flexibility of the humoral immune response against a semiconserved sequence (YX(44)LFX(47)KEKMX(52)L) of the key malaria blood stage vaccine candidate merozoite surface protein-1 (MSP-1). Monoclonal antibodies (mAbs) raised against one of the six described natural sequence variants of MSP-1(43-53) were analyzed for cross-reactivity with the other allelic forms, which differ in one to three positions from the immunizing sequence. Enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR) spectroscopy demonstrated marked differences in mAb binding avidity to the variant sequences and isothermal titration calorimetry (ITC) provided evidence for a very low affinity of some of the interactions. In immunofluorescence analysis (IFA) and Western blotting analysis, the mAbs nevertheless stained all analyzed parasite clones expressing MSP-1(43-53) variant sequences. When used for the evaluation of humoral immune responses in clinical malaria vaccine trials, these two commonly used methods may thus not be suitable to distinguish biologically functional high affinity antibody responses from irrelevant low affinity cross-reactivities. PMID- 12738358 TI - Vaccinia assay for the rapid detection of functional HIV-specific CD8+ cytotoxic T lymphocytes. AB - Human immunodeficiency virus (HIV)-specific T cells play a critical role in anti virus immunity. Therefore, during the clinical development of immune-based therapies, it is important to perform a diagnostic test that rapidly quantifies and characterizes cellular immune responses. For detection of functional HIV specific CD8(+) cytotoxic T cells (CTL), we used a rapid vaccinia assay that employs recombinant vaccinia virus-infected peripheral blood mononuclear cells (PBMC). This assay was compared with the traditional 51Cr-release CTL assay and with the peptide pool-based enzyme-linked immunospot assay (ELISPOT). We demonstrated a close correlation between these assays by using identical antigens in parallel assays. However, the vaccinia assay was the least expensive and time- and labor-consuming test. Regarding sensitivity, the vaccinia assay was similar to both the peptide pool-based ELISPOT and CTL assays. We showed that human histocompatibility leukocyte antigen (HLA)-Dr(+) cells are responsible for presenting recombinant vaccinia antigens to T cells. The recombinant vaccinia based assay is a rapid, sensitive and quantitative test and is thus suitable for the detection of functional antigen-specific CD8(+) CTL in large-scale clinical studies. PMID- 12738359 TI - Purification of natural killer cell cytotoxic granules for assaying target cell apoptosis. AB - We compared two methods originally devised to purify cytoplasmic granules from granulocytes for their capacity to produce cytotoxic granules from natural killer cell lines, suitable for use in target cell apoptosis assays. Both methods utilised nitrogen cavitation to efficiently lyse cells, followed by density gradient fractionation on Percoll to separate the granules from other organelles and granule debris. The first method, originally described by Millard and colleagues, employed DNase I to reduce the viscosity of the initial cell lysate, but the resulting granule fractions were found to contain residual nuclease activity that made them unsuitable for use in apoptosis assays that measure DNA fragmentation. An alternative method described by Borregaard and colleagues utilised a cell relaxation buffer without DNase I. Cytotoxic granules isolated from the NK tumor cell line YT by this protocol were localised predominantly to the densest Percoll fractions, with a density of approximately 1.13 g/ml. These granule fractions were rich in perforin and enzymatically active granzyme B, and induced potent Ca(2+)-dependent lysis and DNA fragmentation of Jurkat cells. Corresponding fractions from non-cytolytic cells, or YT granule extracts incubated with EGTA were unable to mediate significant target cell damage. Cytotoxic granule extracts purified through the Borregaard method were therefore free of nonspecific nuclease activity, and most suitable for studying the mechanism of target cell death induced through the perforin/Ca(2+)-dependent granule pathway. PMID- 12738360 TI - Immune monitoring in whole blood using real-time PCR. AB - There is a need for simple and sensitive assays to assess innate and adaptive immune responses to microbial agents and vaccines. Herein, we describe a whole blood method allowing to measure the induction of cytokine synthesis at the mRNA level. The originality of this method consists in the combination of PAXgene tubes containing an mRNA stabilizer for blood collection, the MagNA Pure instrument as an automated system for mRNA extraction and RT-PCR reagent mix preparation, and the real-time PCR methodology on the Lightcycler for accurate and reproducible quantification of transcript levels. We first demonstrated that this method is adequate to measure the induction of interleukin (IL)-1beta and IL 1 receptor antagonist (IL-1 RA) mRNA upon the addition of bacterial lipopolysaccharide (LPS) to whole blood. We then showed that this approach is also suitable to detect the production of mRNA encoding T cell-derived cytokines in whole blood incubated with tetanus toxoid as a model of in vitro immune response to a recall antigen. Finally, we demonstrated that this methodology can be used successfully to assess inflammatory as well as T cell responses in vivo, as it allowed to detect the induction of IL-1beta and IL-1 RA after injection of LPS in healthy volunteers, and also the induction of IL-2 upon recall immunisation with tetanus vaccine. PMID- 12738361 TI - Characterisation of functional biotinylated TNF-alpha targeted to the membrane of apoptotic melanoma cells. AB - The physiologic clearance of apoptotic cells prevents inflammation at the site of cell death and limits the immunogenicity of tumors. In this study we report the functional characterisation of biotinylated tumor necrosis factor-alpha (TNF alpha) after anchorage to apoptotic melanoma cells via a biotin-avidin-biotin bridge. Flow cytometric and morphological analysis showed that biotinylated TNF alpha efficiently bound to apoptotic membrane blebs of dying cells. Membrane bound TNF-alpha (12 fg/cell) killed sensitive WEHI164 cells 250-fold more effectively than equivalent amounts of the soluble cytokine. Furthermore, macrophages engulfing apoptotic cells with membrane-bound TNF-alpha secreted significantly higher amounts of soluble TNF-alpha and lower amounts of interleukin-10 (IL-10). Therefore the bridging of TNF-alpha potentiates its biological function and influences the outcome of the phagocytic clearance of apoptotic tumor cells. PMID- 12738362 TI - A novel cytotoxicity assay to evaluate antigen-specific CTL responses using a colorimetric substrate for Granzyme B. AB - We have utilized the unique enzymatic properties of a key cytotoxic mediator in target cell destruction, Granzyme B (GrB), to establish an attractive alternative to 51Cr-release assays for the assessment of antigen-specific CTL responses. A number of potential colorimetric peptide substrates were compared to evaluate levels of GrB activity in cytolytic cells. The most specific and sensitive substrate for GrB was Ac-IEPD-pNA, as shown by the minimal enzymatic hydrolysis in apoptotic Jurkat cells and strong hydrolysis in human NK cells. When human peripheral blood lymphocytes were stimulated in vitro, elevated GrB levels were detected by both Ac-IEPD-pNA and a GrB ELISA. Analysis of allo-antigen-specific murine CTLs revealed that GrB exocytosis was only detectable upon challenge with appropriate allogeneic target cells and strongly correlated to 51Cr-release data. The validity of using Ac-IEPD-pNA in vaccine trials was demonstrated in mice immunized with allogeneic P815 cells, where GrB enzymatic activity was measurable in ex vivo splenocytes cell cultures only upon co-incubation with P815 targets. Additionally, influenza-infected mice were also assessed for GrB activity following in vitro peptide-stimulation of splenocytes and strongly reflected both peptide-specific tetramer staining and 51Cr-release results. The novel cytotoxic assay presented here should give investigators a sensitive, cross-species, nonradioactive alternative to 51Cr-release assays as a means to assess antigen specific CTL responses in vaccine trials. PMID- 12738363 TI - Stimulation of tumor-reactive T lymphocytes using mixtures of synthetic peptides derived from tumor-associated antigens with diverse MHC binding affinities. AB - The use of reverse immunology may be necessary to identify new tumor-associated antigens, particularly for cancers, against which tumor-reactive T cell populations have been difficult to establish. One approach has been to screen peptides derived from a candidate antigen with high major histocompatibility complex (MHC) binding affinities for the induction of tumor-reactive T lymphocytes in vitro. However, many candidate antigens that are overexpressed in tumors are nonmutated self-proteins, and unlike foreign or mutated proteins, immunodominant epitopes may not be expressed at high density on the surface of tumor cells. Therefore, to identify tumor-associated epitopes, it may be necessary to screen large panels of peptides with wide ranges of MHC binding affinities. The current methodology of stimulating peripheral blood lymphocytes (PBL) from donors expressing the MHC molecule of interest with individual peptides is impractical for screening such large panels. Therefore, we evaluated the use of mixtures of peptides with variable MHC binding affinities for the induction of tumor-reactive T lymphocytes with the melanoma antigens gp100 and an alternate isoform of tyrosinase-related protein 2 (TRP2-6b) as models. A mixture of 10 known human leukocyte antigen (HLA)-A*0201-restricted peptides from gp100 induced melanoma-reactive cytotoxic T lymphoycte (CTL) from multiple patients with metastatic melanoma. The majority of these T cell populations recognized the known immunodominant epitopes gp100:209-217 and gp100:280-288, even though the HLA-A*0201 binding affinities of these peptides were much lower than other peptides in the mixture. Similarly, melanoma-reactive CTL were generated with a mixture of HLA-A*0201-restricted peptides from TRP2-6b, and these responses were directed against the previously identified tumor-associated epitopes TRP2-6b:180 188, TRP2-6b:288-296 and TRP2-6b:403-411. These results suggest that the use of peptide mixtures may facilitate the identification of new tumor-associated antigens through the application of reverse immunology. PMID- 12738364 TI - Semiquantitative and qualitative assessment of B-lymphocyte V H repertoire by a fluorescent multiplex PCR. AB - We established a new tool to perform semiquantitative and qualitative screening for V(H) gene usage frequency during IgH rearrangements in human B-lymphocytes. In two separate multiplex PCRs, the rearranged VDJ regions were amplified with V(H) family-specific primers labeled with different fluorescent dyes (FAM, HEX, NED, or ROX). The relative amount of each of the particular V(H) family products and their ratios were determined by fragment analysis on a ABI PRISM 377 sequencer. We verified that the fluorescent multiplex PCR (FMPCR) shows high specificity and sensitivity, acceptable reproducibility and reliability. Data obtained were well in agreement with results revealed by sequencing following single-cell PCR. Ten healthy volunteers showed a comparable semiquantitative V(H) family distribution. The FMPCR also correctly detected a monoclonal peak in a CLL patient. Thus, labeling primers with various fluorescent dyes allows for an assessment of V(H) family usage and an immediate determination of the involved V(H) gene family if any clonal peaks are present. This method provides a quick, easy, and reliable tool for V(H) repertoire screening of larger populations of patients suffering from diseases with changes in the V(H) repertoire allowing for selection of cases worth a more detailed and cumbersome sequence analysis later on. PMID- 12738365 TI - Unexpected observation of concentration-dependent dissociation rates for antibody antigen complexes and other macromolecular complexes in competition experiments. AB - The dissociation rate constant of bimolecular complexes between macromolecules (k(off)) is often measured in solution by competition experiments and is generally expected to follow first-order kinetics.When measuring k(off) constants by competition for three complexes of high-affinity recombinant antibody fragments with the cognate antigen and for one calmodulin/peptide complex, a surprising dependence between apparent dissociation rate and concentration of competitor (antigen or calmodulin-binding peptide) was observed. Our results may be characteristic for macromolecules consisting of two domains (such as single chain Fv fragments) and may reflect a transient opening of the two domains which are involved in the binding reaction, and which are connected by a polypeptide linker. PMID- 12738366 TI - Peptabodies as tools to test ligands isolated from phage-displayed peptide libraries. AB - We have previously isolated filamentous bacteriophages, expressing linear hexa peptides and homing to bone marrow endothelium (BME), by panning in vivo of a phage-displayed peptide library in mice. Here, we used peptabody fusion proteins to test the binding capacity of the hexa-peptide SSLTTG to BME cells in vitro. To display this motif in a multimeric form, as originally presented on the bacteriophage, we expressed it N-terminally as a fusion with the peptabody cartilage oligomeric matrix assembly protein (COMP) pentamerization domain, either alone or followed by the N1 domain of the pIII phage coat protein. Binding of the peptabody constructs to the mouse BME cell line STR-10 was investigated by immunofluorescence using anti-COMP antibodies. Only peptabody fusion proteins co expressing pIII-N1 exhibited binding to STR-10, regardless of the presence or absence of SSLTTG. These results indicate that the phage coat protein pIII-N1 domain is the principle determinant responsible for the binding of filamentous bacteriophages to cells of the reticulo-endothelial system (RES). Peptabodies expressing pIII-N1 did not bind to the osteoblast-like cell line MC3T3-E1, indicating that binding is mediated by receptors specifically expressed by BME cells in vivo. Polyinosinic acid (poly-I) was able to inhibit binding of bacteriophages and pIII-N1 expressing peptabodies to STR-10, confirming our previous studies showing that bacteriophages bind to scavenger receptors (SR) expressed by BME cells. In summary, the present study shows the usefulness of peptabodies as a general tool to test the binding capacity of peptide ligands identified by phage display. PMID- 12738367 TI - Mannose-binding lectin: comparison of two assays for the quantification of MBL in the serum of pediatric patients. AB - Individuals with mannose-binding lectin (MBL)-deficiency are at an increased risk from infections with mannose-bearing microorganisms. We have investigated two quantitative research assays for measuring MBL protein in serum for routine diagnosis. The evaluation of 817 serum samples with a nephelometric assay revealed two deficiencies, a number far below the postulated 5-10% of the population. Reevaluation of 102 serum samples with an MBL-ELISA detected low levels in 27 cases (26.4%) and clear deficiencies in 21 samples (20.4%). In our hands, the MBL-ELISA permitted the detection of decreased levels of MBL in serum, as occurs in individuals with homozygous or heterozygous MBL gene mutations; in contrast, the nephelometric assay appeared to be unsuitable for the detection of MBL deficiencies. We support the routine measurement of MBL in serum, especially in children with frequent infections. PMID- 12738368 TI - Assays for human mannose-binding lectin. PMID- 12738369 TI - Production of a recombinant anti-morphine-3-glucuronide single-chain variable fragment (scFv) antibody for the development of a "real-time" biosensor-based immunoassay. AB - A recombinant single-chain variable fragment (scFv) antibody to morphine-3 glucuronide (M3G) was produced using genetic material obtained from the spleen cells of mice immunised with a morphine-3-glucuronide-bovine serum albumin (M3G BSA) conjugate. Immunoglobulin light (V(L)) and heavy (V(H)) chain genes were amplified and cloned into pAK vectors for generation of recombinant antibody fragments in Escherichia coli. A competition ELISA assay was developed in PBS to characterise the ability of the antibody fragments to recognise free drug and the detection limits were found to be as low as 3 ng ml(-1). Surface plasmon resonance-based inhibition immunoassays were developed. The recombinant antibody was pre-incubated with various concentrations of free drug followed by injection over a morphine-3-glucuronide-thyroglobulin (M3G-THY) immobilised surface. The response of antibody binding to the surface of the chip was inversely proportional to the amount of free drug in solution. Regeneration conditions for antibody binding to the surface were optimised resulting in a binding regeneration capacity of at least 30 cycles. The inhibition assay for M3G was tested with assay ranges between 3 and 195 ng ml(-1) and 3 and 97 ng ml(-1) in PBS and urine, respectively. PMID- 12738370 TI - Identification of the antigenic determinants of the American cockroach allergen Per a 1 by error-prone PCR. AB - The group I allergen of cockroach is found in both American and German cockroaches, designated as Per a 1 and Bla g 1, respectively. Members of these allergens so far identified are composed of tandem repeats that may cause the high allergenicity of Per a 1 allergen. In this study, we used monoclonal antibodies HW-8 and HW-19, which can inhibit the binding of patient IgE to Per a 1 allergen, to define the structure of the antigenic determinants in Per a 1.0103 (designated C3), an isoallergen of Per a 1 allergen. Two recognition sites are present, one in the N-terminus (aa 1-208) and the other in the C-terminus (aa 208 395). The N-terminal epitope is not accessible to antibody molecules on the pET expressed C3 protein. The C-terminal epitope was further localized to the aa 267 354 region (C3E) by colony immunoscreening of the cDNA epitope library. By negative screening of the mutated C3E expression library generated by error-prone PCR (ER-PCR), an approach which has rarely been applied in epitope mapping, the functional epitope was identified to lie in aa 318-337 with aa 323-331 being the core motif. The minimal region of the functional epitope was further delineated, by sequence alignment, to be D-x-[I, L]-A-[I, L]-L-P-V-D-E-[L, I]-x-A-[L, I], where x represents any amino acids. This motif is found in all Per a 1 allergens and may serve as a basis for designing a peptide vaccine for allergen-specific immunotherapy. To our knowledge, this is the first report for (1) detailed mapping of the cockroach allergens and (2) use of error-prone PCR random mutagenesis and negative selection in molecular allergology. PMID- 12738371 TI - Plaque reduction test: an alternative method to assess specific antibody response to pIII-displayed peptide of filamentous phage M13. AB - Phage-displayed peptide systems have been used to identify the immunogenic epitopes and to develop the design of peptide-based or peptide-displaying phages themselves as vaccine candidates. To estimate the humoral immunity of phage-based vaccine, it is necessary to evaluate the antibody response specifically directed at the displayed peptide. Enzyme-linked immunosorbent assays (ELISAs) and Western blot analysis are commonly used for this purpose. However, using these methods, it is not easy to distinguish the antibody response against phage coat protein or the antibody response specific to the displayed peptide. The purified anti Mycoplasma hyopneumoniae IgG was used to screen heptapeptides displaying on the pIII coat protein of M13 phage. Four selected phage clones were chosen to immunize mice. In order to evaluate the specific antibody response that is directed against heptapeptides, advantage was taken of the natural property of M13 phage to infect Escherichia coli, which is mediated by the pIII coat protein binding with the F pili of E. coli, and plaque reduction tests were performed to assess the specificity of antibody response. By comparing the number of plaques produced by the different phages (which are the same except for the displayed peptides) neutralized by the antiserum, we could demonstrate that the specificity of antibody response is directed against the peptide displayed on pIII coat protein. The results described here indicate that plaque reduction test is a convenient and more precise method to detect the antibody against the phage displayed peptide. PMID- 12738372 TI - Isolation and expression of recombinant antibody fragments to the biological warfare pathogen Brucella melitensis. AB - Brucella melitensis is a highly infectious animal pathogen able to cause a recurring debilitating disease in humans and is therefore high on the list of biological warfare agents. Immunoglobulin genes from mice immunized with gamma irradiated B. melitensis strain 16M were used to construct a library that was screened by phage display against similarly prepared bacteria. The selected phage particles afforded a strong enzyme-linked immunosorbent assay (ELISA) signal against gamma-irradiated B. melitensis cells. However, extensive efforts to express the respective single chain antibody variable region fragment (scFv) in soluble form failed due to: (i) poor solubility and (ii) in vivo degradation of the c-myc tag used for the detection of the recombinant antibodies. Both problems could be addressed by: (i) fusing a human kappa light chain constant domain (Ck) chain to the scFv to generate single chain antibody fragment (scAb) antibody fragments and (ii) by co-expression of the periplasmic chaperone Skp. While soluble, functional antibodies could be produced in this manner, phage-displaying scFvs or scAbs were still found to be superior ELISA reagents for immunoassays, due to the large signal amplification afforded by anti-phage antibodies. The isolated phage antibodies were shown to be highly specific to B. melitensis and did not recognize Yersinia pseudotuberculosis in contrast to the existing diagnostic monoclonal YST 9.2.1. PMID- 12738373 TI - Detection of H-2 alleles by PCR-RFLP-an alternative to flow cytometry in the screening of transgenic mice. AB - An increasing number of experimental models is based on well-defined transgenic mice in medical and biological research. Particularly in settings in which transgenic recombinants are used, a fast and reliable method is needed to screen for a defined H-2 background. For this purpose, flow cytometry with specific monoclonal antibodies is the standard procedure. However, epitopes of closely related rodent strains show only minor variations affecting the production of specific discriminating antibodies. Therefore, cross-reactivity of antibodies against specific major histocompatibility complex (MHC) leads to unreliable results in settings with closely related strains. In need of a method with high reliability, we have designed a screening assay based on polymerase chain reaction (PCR) followed by restriction fragment length polymorphisms (RFLP) to discriminate the MHC class I antigens H-2K(d), -K(b), -K(k), which are sequence variants of the H-2K gene. A part of the mus musculus MHC gene coding for H-2K covering exons 4 and 5 with MHC-differentiating restriction sites-was amplified. Subsequent restriction digest of the PCR products allows to discriminate the three aforementioned alleles and to identify homozygous as well as heterozygous haplotypes. To distinguish transgenic mice defined by certain MHC backgrounds, the PCR-RFLP method is simple, cost-effective, specific, and reliable and can be used independently or in addition to other methods in any laboratory. PMID- 12738374 TI - Differential expression of IFN-alpha subtypes in human PBMC: evaluation of novel real-time PCR assays. AB - Studies of the human IFN-alpha subtype system have been hampered by the lack of efficient procedures to quantify and differentiate the expression of the highly homologous IFN-alpha subtypes. Here we evaluate four novel real-time PCR assays for the specific detection and quantification of IFN-alpha mRNA for the subtypes alpha(2), alpha(6), alpha(8) and alpha(1/13) in a combined assay in human peripheral blood mononuclear cells (PBMC). This included (a) the selection of beta-glucuronidase (GUS) as a suitable housekeeping gene for relative quantification; (b) verification of the specificity by using human DNA of different IFN-alpha subtypes; and (c) comparison of the amplification efficiencies among the different assays. This highly sensitive method allows the detection of low-level, constitutive IFN-alpha mRNA and shows differences in the composition of constitutive IFN-alpha subtypes compared to other cell types (HeLa and HEp-2). The in vitro stimulation of PBMC with Newcastle disease virus (NDV), Respiratory syncytial virus (RSV) or an inactivated Herpes simplex (HSV) preparation leads to the transcriptional induction of all IFN-alpha subtypes investigated but to different expression levels. Among the subtypes detected, IFN alpha(13/1) and alpha(2) are the major transcripts followed by alpha(8), and finally alpha(6) as a minor transcribed subtype. Time-kinetics of IFN-alpha transcriptional activation also revealed variations in the course of IFN-alpha transcription between NDV, RSV or HSV. The data obtained from the real-time PCR assays correlated well with IFN-alpha(2) protein release. In conclusion, we have demonstrated the suitability and reliability of new real-time PCR assays for the rapid and efficient analysis of IFN-alpha subtype expression. PMID- 12738375 TI - Double-blotting: a solution to the problem of nonspecific binding of secondary antibodies in immunoblotting procedures. AB - "Double-blotting" (DB) has been developed to overcome the problem of nonspecific binding of secondary antibodies in immunoblotting (IB). After it has been probed by the primary antibody, the membrane with the blotted proteins is assembled with a second blank membrane and submitted to a second blotting under acidic conditions. The primary antibody molecules are thus desorbed from their corresponding antigen and transferred onto the second membrane, whereas the antigen and the interfering proteins remain bound to the first one. The second membrane can then be probed by the secondary antibodies without the risk of nonspecific binding. This method has been developed for the study of erythropoietin (EPO) in concentrated urine since a strong nonspecific binding of biotinylated secondary antibodies to some urinary proteins is observed using classical IB protocols. However, its concept makes it usable in other applications that come up against this kind of problem. PMID- 12738377 TI - A simple and rapid vortex method for preparing antigen/adjuvant emulsions for immunization. AB - The sustained release of antigen over a period of time is necessary to stimulate the immune system and generate a high titer of antibody. The use of adjuvants in water-in-oil emulsions has long been the favored method of prolonging the exposure time of antigens to enhance the immune response. Routine methods for preparation of this emulsion are repeated mixing of the materials in a "two syringe" method that is labor-intensive and unreliable. The method we describe here is to use a common vortex (i.e. Vortex Genie 2, VWR) to prepare water-in-oil antigen/adjuvant emulsions. This method has been routinely used in our laboratory for many years and demonstrates its consistency with reliability for immunization of mice, hamsters, and rats and subsequently generation of hybridomas. PMID- 12738376 TI - EliCell assay for the detection of released cytokines from eosinophils. AB - Eosinophils contain several preformed cytokines within their specific granules. Therefore, without requiring them in de novo synthesis of cytokines, eosinophils can release quantities of granule-derived cytokines by highly regulated mechanisms. However, eosinophil "degranulation" is poorly understood, in part, because available methodologies did not appear appropriate for analyzing vesicular mobilization and transport of eosinophil granular contents. The EliCell assay is a microscopic methodology substantially modified from other techniques employed to detect cytokine release (i.e., ELISPOT). The method is a dual antibody capture/detection system in which viable eosinophils are incubated in a solid streptavidin-conjugated agarose matrix, which contains a biotinylated capture antibody against the cytokine of interest. Released cytokine is detected around non-permeabilized eosinophils with a separate fluorochrome-labeled detection antibody. Thus, the EliCell system captures and detects extracellular cytokines at the site of their release from eosinophils. As examples, we have used EliCell essays to detect the selective release of either IL-4 or IL-12 cytokines found preformed in eosinophils-from eotaxin- or anti-CD9-stimulated eosinophils, respectively. With appropriate pairs of antibodies, any preformed cytokine found into eosinophil granules could be studied and the mechanisms of their secretion evaluated by using the EliCell assay. PMID- 12738379 TI - Helicobacter pathogenesis and immunology. Papers from the 5th International Workshop on Pathogenesis and Host Response in Helicobacter Infections. Helsingor, Denmark. July 2002. PMID- 12738380 TI - Helicobacter pylori: an invading microorganism? A review. AB - In this review we evaluate the pros and cons of Helicobacter pylori invasion of epithelial cells as part of the natural history of H. pylori infection. H. pylori is generally considered an extracellular microorganism. However, a growing body of evidence supports the controversial hypothesis that at least a subset of H. pylori microorganisms has an intracellular (intraepithelial) location. Most significant is the fact that H. pylori invades cultured epithelial cells with invasion frequencies similar to Yersinia enterocolitica and better than Shigella flexneri; furthermore, studies of invasion mechanisms suggest that H. pylori invasion of and survival within epithelial cells is not merely a passive event, but requires active participation of the microorganism. Although many studies of human gastric biopsy specimens have failed to demonstrate any intracellular H. pylori, some studies have revealed a minor fraction of H. pylori inside gastric epithelial cells, with possible linkage to peptic ulceration and epithelial cell damage. In conclusion, these data encourage further research to establish whether intracellular H. pylori does play a role in H. pylori colonization of the human stomach and in peptic ulcer pathogenesis. PMID- 12738381 TI - Interactions of the gastrotropic bacterium Helicobacter pylori with the leukocyte endothelium adhesion molecules, the selectins--a preliminary report. AB - The deleterious effects of Helicobacter pylori infection of the stomach are largely the result of a vigorous chronic inflammatory response, and include chronic gastritis, peptic ulceration and gastric cancer. We are exploring the possibility that carbohydrate components on H. pylori contribute to the persistent inflammation through interactions with leukocyte-endothelial adhesion molecules of the host. Lipopolysaccharides of most H. pylori strains contain sequences related to the Lewis (Le(x) or Le(a)) antigens. Carbohydrate sequences of this family encompass ligands for the leukocyte-endothelium adhesion molecules of the host, namely, the E- and P-selectins, which are expressed on inflamed endothelia, and L-selectin, which is constitutively expressed on leukocytes. Here we investigate H. pylori isolates from patients with chronic gastritis, duodenal ulcer and gastric cancer for their interactions with the selectins. Our results provide unequivocal evidence of interactions of isolates from each of the diagnostic groups with E- and L-selectins. PMID- 12738382 TI - Investigation of the biological relevance of Helicobacter pylori cagE locus diversity, presence of CagA tyrosine phosphorylation motifs and vacuolating cytotoxin genotype on IL-8 induction in gastric epithelial cells. AB - Isolates of Helicobacter pylori from dyspeptic patients in England and South Africa were tested for ability to induce interleukin-8 (IL-8) in gastric cells. All isolates were cagA-positive, which was used as a marker for the presence of the cag pathogenicity island. The aims were to determine if activities were related to diversity within cagE (HP0544), a locus encoding a key component in the Type IV secretion system, and if disease severity might be linked to a combination of strain features. We found that isolates were heterogeneous in ability to induce IL-8 activity with the 23 positive isolates (59%) showing activities ranging from 260 to 3200 pg ml(-1). The cagE locus was detected in most isolates and RFLP analysis of a 1.52-kb internal fragment showed interstrain diversity with 12 combined (MboI/NlaIII) types. Most cagE genotypes were not associated with IL-8 induction, however two genotypes were found only in IL-8 inducing strains and one genotype was associated with lack of IL-8 induction. IL 8 activity was not associated with either the number or composition of cagA tyrosine phosphorylation motifs and vacA m-type. Although we found a weak association between cagE type and the ability to induce IL-8, our results imply that gastric cell factors or bacterial factors other than vacA, cagA and cagE are involved in the induction of IL-8 and the development of severe gastric disease. PMID- 12738383 TI - Helicobacter pylori lipopolysaccharide in the IL-2 milieu activates lymphocytes from dyspeptic children. AB - In this study, we assessed the proliferative response of peripheral blood mononuclear leukocytes (PBML) from 33 children/young adolescents with chronic dyspepsia, to H. pylori LPS in the presence and absence of IL-2 as a T cell growth factor. A rapid urease test (RUT) and a presence of Helicobacter-like organisms (HLO) in the biopsy specimens allowed us to distinguish RUT/HLO positive (17/33) and -negative (16/33) patients. H. pylori LPS alone induced a proliferation of PBML from 4 out of 33 dyspeptic patients. IL-2 increased the prevalence of the response to LPS to 59% and 74% of RUT/HLO-positive and negative patients, respectively. PBML from RUT/HLO-positive patients responded significantly less intensively to H. pylori LPS in the presence of IL-2, to IL-2 alone and to H. pylori LPS+IL-2. However, there was no difference in PHA-driven proliferation of PBML from the patients of those two groups. A negative correlation between the responsiveness to H. pylori LPS of PBML and occurrence of type B inflammation in gastric mucosa was demonstrated. The results suggest a contribution of H. pylori LPS to an outcome of H. pylori infection. It is speculated that H. pylori LPS by an activation of immunocompetent cells may reduce gastric inflammation, decrease bacterial load and prolong H. pylori infection. PMID- 12738384 TI - The cagA status of Helicobacter pylori isolates from dyspeptic children in Turkey. AB - There are inconsistent reports regarding cytotoxin-associated gene A (cagA) status of Helicobacter pylori isolates and the severity of the mucosal lesions in children. The aim of this study was to determine the prevalence of cagA(+) strains and to evaluate its correlation with clinic and endoscopic findings. We examined 45 H. pylori strains that were grown on brain-heart infusion agar supplemented with 7% horse blood. Following 72 h of incubation colonies were harvested and bacterial DNA was extracted. Polymerase chain reaction primers F1 and B1 were used to amplify a 348-bp internal fragment of cagA. The prevalence of cagA in Turkish pediatric patients was 55.6%. No association was found between cagA status and the severity of gastro-duodenal lesions. PMID- 12738385 TI - Potential for proteomic profiling of Helicobacter pylori and other Helicobacter spp. using a ProteinChip array. AB - The Helicobacter genus is associated with a wide spectrum of pathologies in the gastrointestinal tract. However, in contrast to Helicobacter pylori, few data are available regarding proteomic characteristics of enterohepatic helicobacters. Proteomic analysis of this genus has predominantly utilised two-dimensional gel electrophoresis methodology. In the present study we applied an innovative technique using ProteinChip arrays coupled with surface-enhanced laser desorption/ionisation time of flight mass spectroscopy to accurately assess the M(r) of proteins for comparative proteomic profiling. We analysed binding of outer membrane fractions to a weak cation exchange array for strains of H. pylori from culture collections and compared these profiles to fresh clinical isolates. In addition, we analysed profiles from Helicobacter pullorum, Helicobacter bilis and 'Helicobacter sp. flexispira'. The system proved rapid, accurate and reproducible. Distinct specific profiles for all the strains studied were identified. However, strains from culture collections that have undergone numerous subcultures had almost identical profiles. In contrast, profiles from fresh clinical isolates were markedly different. Moreover, certain features of the profiles from the enterohepatic species were conserved. PMID- 12738386 TI - Increased frequency of activated T-cells in the Helicobacter pylori-infected antrum and duodenum. AB - Helicobacter pylori colonize the human stomach and duodenum. The infection has been shown to induce a strong T-cell response in the stomach, whereas the response within the duodenum has been poorly characterized. Furthermore, it remains to be elucidated whether the T-cell response may contribute to ulcer formation in the host. In this study, the frequency of different T-cell subsets, their degree of activation and expression of co-stimulatory receptors in biopsies from the duodenum as well as the antrum were studied by immunohistochemistry and flow cytometry. It was also evaluated whether there are differences in the T-cell responses between duodenal ulcer patients and asymptomatic carriers that might explain why only 10-15% of the infected subjects develop duodenal ulcers. The frequencies of CD4+, CD8+ and CD45RO+, i.e. memory T-cells, were significantly increased in the antrum, and the number of CD25+ cells was considerably higher in both the antrum and duodenum of duodenal ulcer patients and asymptomatic carriers as compared to uninfected individuals. Interestingly, the levels of immunosuppressive CTLA-4+ cells were significantly higher in the duodenum of duodenal ulcer patients, as compared to the asymptomatic carriers. H. pylori cause activation of T-cells in the duodenum as well as in the stomach. Our observation of higher levels of CTLA-4+ cells in the duodenum of duodenal ulcer patients than in the asymptomatic carriers suggests that a suppressive T-cell response may be related to the development of duodenal ulcers. PMID- 12738387 TI - Orogastric vaccination of guinea pigs with Helicobacter pylori sonicate and a high dose of cholera toxin lowers the burden of infection. AB - Guinea pigs were vaccinated orogastrically with Helicobacter pylori cell sonicate (CS) and 10 microg or 100 microg cholera toxin (CT) or CT only. Nai;ve animals were used as a control. In both experiments, vaccination primed the local IgG and IgA response, irrespective of the CT dose. After challenge, only the group of animals immunised with CS and 100 microg CT had a significantly lower number of H. pylori in the antral region of the stomach, but vaccination did not prevent H. pylori infection. This protective effect was not associated with a switch in IgG subclass, which remained predominantly IgG2. The levels of specific antibodies in serum and the gastric mucosa which were similar to naive unprotected animals. In conclusion, the ability of mucosal adjuvants such as CT to induce a protective immune response may be host dependent and findings in the Helicobacter-mouse model should be interpreted with caution. PMID- 12738388 TI - Gastric mucosal cytokine responses in Helicobacter pylori-infected patients with gastritis and peptic ulcers. Association with inflammatory parameters and bacteria load. AB - Helicobacter pylori is an important pathogen in gastroduodenal inflammation and ulceration. Several mechanisms have been proposed to explain its role. We studied the cytokine production patterns in situ in gastric mucosal biopsies from H. pylori-positive and H. pylori-negative patients with dyspepsia. Immunohistochemistry with monoclonal antibodies was used. The study showed enhanced expression of interleukin (IL) -8, IL-10 and interferon-gamma (IFN gamma) in H. pylori infection and a significant association was found between these cytokines and the following parameters: bacteria load, chronic inflammation and activity. These parameters were significantly correlated with the cell markers CD19 and CD56. The study indicates a dual effect of H. pylori on the Th1 response, i.e. a stimulation of the response verified by increased IFN-gamma and a feed-back verified by an increase of the counterinflammatory IL-10, which may dampen the inflammatory and cytotoxic effect of the Th1 response. Furthermore, the study confirms the connection between increase of IL-8 and inflammatory activity in gastric mucosa in H. pylori infection. PMID- 12738389 TI - Clinical relevance of the cagA, vacA and iceA genotypes of Helicobacter pylori in Brazilian clinical isolates. AB - Infection with Helicobacter pylori strains harboring determinants of pathogenicity may lead to a strong inflammatory response in gastric mucosa. In this work, we examined the frequency of the cagA, vacA and iceA genotypes in H. pylori strains isolated from Brazilian patients and correlated these with the clinical manifestations. H. pylori was isolated from 165 patients [30 with non ulcer dyspepsia cases (NUD); 93 peptic ulcer disease (PUD): 31 gastric ulcers (GU) and 62 duodenal ulcer disease (DU); 18 with erosive gastritis (EG); and 24 gastroesophageal reflux disease (GERD)]. Allelic variants of cagA, vacA and iceA were identified using the polymerase chain reaction. More than one H. pylori strain was detected in 28 cases (17%), and these were excluded from the statistical analysis. We were unable to confirm an association between iceA status and clinical outcome. There was a strong association between the genotype cagA-positive vacA s1 and PUD. However, logistic regression analysis showed that vacA s1 was the only predictive factor for PUD (OR=4.19; 95% CI 1.95-8.98). The presence of the less virulent strain vacA s2 was related to GERD (OR=8.59; 95% CI 2.85-25.91). Our results support the hypothesis that virulent strains may protect against the development of GERD. PMID- 12738391 TI - Analysis of the rdxA gene in high-level metronidazole-resistant clinical isolates confirms a limited use of rdxA mutations as a marker for prediction of metronidazole resistance in Helicobacter pylori. AB - Metronidazole (Mtz) resistance in the gastric pathogen Helicobacter pylori is closely associated with inactivation of the nitroreductase gene rdxA. In order to identify respective mutations for diagnostic purposes we analyzed the rdxA gene in a collection of high-level Mtz-resistant clinical H. pylori isolates. Size alterations in the rdxA gene region were found in only two out of 45 and one out of 40 isolates showing lower-level (minimal inhibitory concentrations (MICs) 32 192 microg ml(-1)) and high-level (MIC>/=256 microg ml(-1)) Mtz resistance, respectively. Point mutations that interrupt the rdxA reading frame were detected in two out of eight high-level resistant isolates (MICs>/=256 microg ml(-1)). Most remarkably, the rdxA gene sequence was found to be identical in four out of five high-level Mtz-resistant and -susceptible paired H. pylori isolates from the same patients each. Taken together, these results demonstrate that although some isolates carry classical resistance-associated rdxA mutations, as described earlier, the use of rdxA mutations as a marker for prediction of Mtz resistance is limited. PMID- 12738390 TI - A link between Helicobacter pylori and/or Chlamydia spp. infections and atherosclerosis. AB - Antibodies to Helicobacter pylori, Chlamydia spp. and Mycobacterium bovis were determined in patients with coronary heart disease, H. pylori-related dyspepsia, and tuberculosis, and healthy controls. Enzyme-linked immunosorbent assay was conducted with a glycine extract and CagA protein of H. pylori, chlamydial lipopolysaccharide and mycobacterial heat shock protein Hsp65. The prevalence of anti-glycine extract IgG in coronary heart disease patients was higher than in the tuberculosis group and controls, and the same as in dyspeptic patients. Anti chlamydial IgG were more prevalent in the coronary heart disease group than in healthy subjects. There was no difference in the prevalence of anti-CagA IgG in the coronary heart disease group and controls or anti-Hsp65 IgG in the patients with coronary heart disease, dyspepsia, tuberculosis, and controls. Anti-glycine extract IgA (like anti-glycine extract IgG) were more prevalent in the coronary heart disease group than in the healthy group. The highest anti-glycine extract IgG/IgA and anti-chlamydial IgG titers were more frequent in coronary heart disease patients as compared with controls. Infections with H. pylori and Chlamydia spp. and enhanced production of antibodies to these pathogens may predispose to human atherosclerosis. PMID- 12738393 TI - Infection of Mongolian gerbils with Chinese Helicobacter pylori strains. AB - To date only a few Helicobacter pylori strains have been demonstrated to colonise Mongolian gerbils successfully. The aim of this study was to establish stable colonisation of Chinese strains of H. pylori in gerbils. Fresh clinical isolates from Chinese patients were inoculated into gerbils. At 4-6 weeks post inoculation, infection status was evaluated by culture, biopsy urease test and pathology. Sequencing of glmM and random amplified polymorphic DNA (RAPD) fingerprinting of DNA from cultured H. pylori were used to evaluate the genetic identity of pre-inoculated and post-inoculated strains. The ability of pre- and post-inoculated strains to stimulate interleukin-8 transcription in L5F11 gastric epithelial cells was analysed. Three of five clinical isolates colonised gerbils. The three pre- and post-inoculation strains had identical glmM sequences and RAPD profiles, and stimulated luciferase secretion from L5F11 epithelial cells. The strain that caused severe pathological changes was selected for repeat infection to prove reproducible and stable colonisation. The cagA+ strain 42GX gave stable colonisation in the gerbil and induced severe gastritis. PMID- 12738392 TI - Differences in immunogenicity and protection in mice and guinea pigs following intranasal immunization with Helicobacter pylori outer membrane antigens. AB - Mice and guinea pigs were intranasally immunized with either recombinant lipoprotein 20 or Helicobacter pylori outer membrane vesicles (OMV). Cholera toxin was used as mucosal adjuvant. In mice, both vaccines elicited systemic and local IgG responses, which correlated with significantly lower levels of H. pylori colonization. In contrast, only OMV proved immunogenic in guinea pigs, with the development of both systemic and local immune responses. These antibodies did not, however, correlate with protection in these animals, which suggests that vaccine formulation is as important as choice of antigen in the development of an H. pylori vaccine. PMID- 12738394 TI - Diagnostic criteria for gestational diabetes in relation to pregnancy outcome. AB - OBJECTIVES: To determine which of the American Diabetes Association (ADA) or World Health Organization (WHO) plasma glucose criteria for gestational diabetes mellitus (GDM) best predicts poor fetal outcome. To determine whether an alternative cut-off point would result in increased predictive value and greater diagnostic effectiveness in pregnancies at high risk for GDM. RESEARCH DESIGN AND METHODS: A sample of 473 successive apparently normal pregnant women attending the Obstetric Department were screened for GDM using both the ADA and the WHO criteria. Between 26 and 30 weeks of gestation, they underwent, on subsequent days, a screening test with a 50-g oral glucose load and two oral glucose tolerance tests (OGTTs) with 75 and 100 g of glucose according to the WHO and the ADA recommendations, respectively. From this group, we identified 99 women at high risk for GDM, who did not attend their pregnancy follow-up and whose delivery records were recovered at our hospital or in neighbouring hospitals. This unusual situation enabled us to study the natural history and outcome of their pregnancy in spite of not receiving special management usually provided to such women. As macrosomia was expected to be the most frequent undesirable foetal outcome, sensitivity and specificity calculations have been based on this outcome. RESULTS: The study population (n=99) had a median parity of two and 14% had abnormal results in the 2-h, 75-g load test (WHO) vs. 6% in the 100-g test (ADA). Optimal cut-off points for each test were lower than those recommended for diagnosis by the ADA and the WHO. The optimal sensitivity for the 1-h, 50-g test was 66.7% (cut-off 137 mg/dl), and for the 2-h, 75-g test (cut-off 119 mg/dl). The best specificity and positive predictive value was for this last test with a cut-off point of 140 mg/dl in the second hour. CONCLUSIONS: The standard 2-h cut off value of 140 mg/dl for the 75-g test, as now recommended by WHO, was optimal for predicting macrosomia. Based on the sensitivity and specificity for macrosomia, the 1-h, 50-g screening test had an optimal cut-off point of 137 mg/dl (vs. 140 mg/dl recommended by ADA). The 2-h, 75-g OGTT value using a cut off point of 119 mg/dl had equivalent sensitivity, specificity, and positive predictive value. In contrast, the 100-g OGTT had much lower levels of sensitivity, but higher specificity and higher positive predictive value. PMID- 12738395 TI - Associations between inflammatory markers, traditional risk factors, and complications in patients with type 2 diabetes mellitus. AB - OBJECTIVE: Inflammatory markers predict cardiovascular events in a wide range of patients. Two factors, fibrinogen (FIB) and high-sensitivity C reactive protein (CRP), are currently entering clinical practice as cardiovascular risk predictors. In patients with type 2 diabetes mellitus, we sought to examine the relationship between macrovascular disease, urinary albumin/creatinine ratio (ACR), and FIB or CRP, as well as the relationship of FIB and CRP with traditional risk predictors of these complications of diabetes. METHODS: In 202 consecutive patients with type 2 diabetes mellitus from a diabetes clinic, clinical and biochemical data were obtained and a cross-sectional analysis was performed. RESULTS: Patients with macrovascular disease had higher FIB (P=.02) but not higher CRP. They were older, more likely to have retinopathy or elevated serum creatinine, had higher ACR and lower HDL cholesterol. They were more likely to be treated with statins, beta-blockers, and ASA. Adjustment for statin therapy did not result in significant differences in CRP levels according to macrovascular disease status. Both FIB (P=.01) and CRP (P=.02) were significantly higher in patients with ACR whose values were in the proteinuria range. In multivariate analysis, both FIB (P=.001) and CRP (P=.03) were positively correlated with ACR, but no association was seen between CRP and ACR when FIB was entered in the model. Other factors positively associated with ACR were age, diastolic blood pressure, retinopathy, and hemoglobin A1c (HbA1c). FIB and CRP were strongly correlated (R=.49, P< or =.001) and this effect was independent of statin therapy. CRP was positively associated with body mass index (BMI), serum triglycerides, and sulfonylurea therapy and negatively associated with metformin therapy. Patients on statin therapy had significantly higher FIB and lower CRP. Women on hormone replacement therapy (HRT) had significantly lower FIB and higher CRP. CONCLUSIONS: In patients with diabetes: (1) the two markers, FIB and CRP, are interrelated; (2) FIB is significantly associated with presence of microvascular disease, independent of CRP; (3) CRP is strongly associated with metabolic factors but not with complications of diabetes, independently of FIB; (4) statins and HRT were divergently associated with CRP and FIB as HRT was associated with lower FIB and higher CRP, while statins showed the reverse association; and (5) CRP and FIB provide different information about the characteristics and consequences of diabetes mellitus because of divergent associations with biological indicators and therapeutic agents. PMID- 12738397 TI - Lipoprotein(a), apolipoprotein(a) polymorphism and restenosis after intracoronary stent placement in Type 2 diabetic patients. AB - The relationship between lipoprotein(a) [Lp(a)] and restenosis after intracoronary stent implantation has never been analysed in diabetic patients. The aim of the present prospective study was to evaluate whether Lp(a) levels and apolipoprotein(a) [apo(a)] phenotypes are predictors of restenosis after elective stent implantation in Type 2 diabetic patients with de novo lesions of coronary arteries. We recruited 102 Type 2 diabetic patients with a new lesion successfully treated with elective placement of one or two Palmaz-Schatz stents. Follow-up angiography was scheduled at 6 months or earlier if clinically indicated. Seven patients were lost to the follow up. Among 95 patients enrolled, restenosis was present in 37 (38.9%) and absent in 58 (61.1%). The restenosis group showed Lp(a) levels higher than the nonrestenosis group (25.1+/-14.4 vs. 21.3+/-14.6 mg/dl), but the difference was not significant. The restenosis group had a percentage of subjects with at least one apo(a) isoform of low molecular weight (MW) significantly greater than the nonrestenosis group (75.7% vs. 55.1%; P<.05). A multiple logistic regression analysis showed that presence of multivessel disease (risk relative [RR]: 5.83; 95% confidence interval [CI]: 1.21 28.15; P<.05) was the only predictor of restenosis after stent placement in diabetic patients. Lp(a) and apo(a) polymorphisms did not enter the model as predictive variables. Our study shows that the presence of multivessel disease is a predictor of restenosis after intracoronary stent implantation in diabetic patients. On the contrary, Lp(a) and apo(a) polymorphisms do not appear to be reliable markers of restenosis in patients with Type 2 diabetes mellitus. PMID- 12738396 TI - The relationship between self-monitoring of blood glucose control and glycosylated haemoglobin in patients with type 2 diabetes with and without diabetic retinopathy. AB - BACKGROUND: Diabetes mellitus (DM) is a major health problem with long-term microvascular and macrovascular complications responsible for the majority of its mortality and morbidity. The development and progression of diabetic complications are strongly related to the degree of glycemic control. To decrease the occurrence of these complications, instruments for self-monitoring of blood glucose (SMBG) have been developed and have become widely used among diabetic patients. In this study, we determined the relationship between SMBG control and glycosylated haemoglobin (HbA(lc)) levels in patients with type 2 diabetes, with and without diabetic retinopathy. METHODS: Two hundred and sixty-seven type 2 diabetic patients (mean age [mean+/-S.D.]: 58.07+/-9.13 years, duration of diabetes: 8.63+/-6.8 years) participated in this study. Following an educational program on SMBG, glucometers and usage of oral antidiabetic agents or insulin, optic fundi were examined and HbA(lc) levels were measured at baseline and after 6 and 12 months. The patients were classified in three groups according to their funduscopic findings: without retinopathy (n=140, 52.4%), background retinopathy (n=75, 28.1%) and proliferative retinopathy (n=52, 19.5%). RESULTS: HbA(lc) levels at baseline, after 6 and 12 months were 9.09+/-2.69%, 7.47+/-1.78% and 7.12+/-1.4%, respectively, mean+/-S.D. The values decreased significantly after the education program (P<.001 for both values compared with baseline). The prevalence of retinopathy (both background and proliferative) was 0.8% in the group of diabetics with a mean HbA(lc) level <6%, 7.1% in those between 6.1% and 6.9%; 9.4% in those between 7% and 7.9%; 11.8% between 8% and 8.9%; and 70.9% in those exceeding a mean HbA(lc) level of 9%. There was a statically significant relationship between proliferative diabetic retinopathy and body mass index (BMI; P<.001). The same relationship was observed between duration of diabetes and diabetic retinopathy (P<.001), but not between sex and diabetic retinopathy (P=.46). CONCLUSIONS: Implementing a program of SMBG control in type 2 diabetic patients results in lower levels of HbA(lc) at 6 and 12 months. In the group without diabetic retinopathy at 6- and 12-month controls, the mean HbA(lc) concentration is less than 7%, but in the group with diabetic (background and proliferative) retinopathy, this value could not be reduced below 7%. These results imply that SMBG would allow us to maintain better metabolic control by improving HbA(lc) levels and we have always kept in mind that SMBG was a part of an educational program. On the other hand, improving glycemic control prevents the onset or progression of diabetic microvascular complications, such as diabetic retinopathy, nephropathy and neuropathy. Long-term clinical studies should be performed to determine cost-effectiveness and the effects of SMBG on diabetic complications, morbidity and mortality. PMID- 12738399 TI - Intractable vomiting in diabetic patients. AB - Intractable nausea and vomiting have been described in individuals without any underlying physical etiology explaining these complaints. Physical or emotional abuse has been described in individuals suffering from these symptoms and associated with somatoform disorders manifesting primarily as gastrointestinal complaints. We present five patients with long-standing Type 1 diabetes who suffered from intractable vomiting. Personality disorders, profound depression and emotional abuse dramatically influenced the course of these patients' illness. In most of the patients, physical symptoms remarkably improved after identification and removal of the triggering factors. Therefore, psychogenic vomiting must be considered among the differential diagnoses of intractable nausea and vomiting, especially in individuals with chronic illnesses. A careful search for a physical etiology and medical treatment that does not cause relief of symptoms should suggest that there is almost certainly a psychological issue at the root of the problem. PMID- 12738398 TI - Improvement of esophageal and gastric motility after 2-week treatment of oral erythromycin in patients with non-insulin-dependent diabetes mellitus. AB - Forty-five patients with non-insulin-dependent diabetes mellitus (NIDDM) were enrolled in the study. Before and after the 2-week treatment course of oral erythromycin, esophageal (the mean transit time, MTT) and gastric motility (the halftime of gastric emptying, GETt1/2) were evaluated by radionuclide labeled liquid and solid meals. Meanwhile, the fasting blood sugar (FBS) levels were monitored. After a 2-week erythromycin treatment, MTT shortened from 9.32+/-1.12 to 6.28+/-0.91 s and GETt1/2 shortened from 210.2+/-24.3 to 128.3+/-30.1 min. Meanwhile, FBS decreased from 198.2+/-30.1 to 121.5+/-21.3 mg/dl (P value <.05). We conclude that a 2-week treatment course of oral erythromycin can significantly improve esophageal and gastric motility which results in a better control of blood sugar in patients with NIDDM. PMID- 12738400 TI - Effects of eicosapentaenoic acid on platelet activation markers and cell adhesion molecules in hyperlipidemic patients with Type 2 diabetes mellitus. AB - We compared the levels of microparticles, platelet activation markers, soluble cell adhesion molecules, soluble selectins, and antioxidized low-density lipoprotein (anti-Ox LDL) antibody between patients with hyperlipidemia and control subjects. Binding of anti-glycoprotein (GP) IIb/IIIa and anti-GPIb monoclonal antibodies to platelets did not differ significantly between the hyperlipidemic patients and controls. However, expression of activation markers (CD62P, CD63, PAC-1, and annexin V) by platelets was higher in the hyperlipidemic patients with Type 2 diabetes. The levels of platelet-derived microparticles (PDMPs) and monocyte-derived microparticles (MDMPs) were significantly different in hyperlipidemic patients with Type 2 diabetes and controls. Soluble P-selectin (sP-selectin), soluble E-selectin (sE-selectin), and anti-Ox LDL antibody also showed higher levels in the hyperlipidemic patients with Type 2 diabetes. After treatment with eicosapentaenoic acid (EPA), the levels of CD62P, CD63, annexin V, PDMPs, and MDMPs, sE-selectin, and oxidized LDL antibody were reduced significantly. Triglyceride (TG) and total cholesterol levels were also decreased. Anti-Ox LDL antibodies and MDMPs were correlated positively with platelet CD62P (plt-CD62P) levels. These findings suggest that in hyperlipidemic patients with Type 2 diabetes, EPA may prevent complications caused by oxidized LDL, E-selectin, and activated platelets or monocytes. PMID- 12738401 TI - Evaluation of cystatin C and beta-2 microglobulin as markers of renal function in patients with type 2 diabetes mellitus. AB - BACKGROUND: Despite recent studies showing that serum cystatin C (CC) is a better marker for GFR than the ubiquitously used serum creatinine, its clinical utility remains under evaluation. METHODS: To evaluate their usefulness in patients with type 2 diabetes mellitus (DM), serum concentrations of CC, beta-2 microglobulin (B2M) and creatinine were measured in 105 (38 males, 67 females) Kuwaiti patients with type 2 DM. The results were compared with creatinine clearance (Ccr), which was measured (mCcr) and estimated (eCcr) with the Cockroft-Gault formula, and correlated with 24-h urine protein and early morning urine albumin/creatinine excretion ratio. RESULTS: In patients with eCcr and mCcr results (n=51), eCcr and mCcr showed significant correlation with each other (r's=.86, P<.0001) with no significant difference between the two. In all patients (n=105), CC and B2M showed significant correlation with each other (r's=.82, P<.0001) and with serum creatinine concentration (r's=.77 and.84, respectively, P<.0001). Serum CC, B2M and creatinine showed significant (P<.001) inverse correlation with eCcr (r's= .63, -.61 and -.76, respectively). Partial correlations after correcting for age and sex improved the correlation of serum creatinine with eCcr (r=-.81, P<.0001), but there was no significant change in the correlations of CC and B2M with eCcr (r=-.65, P<.0001 and r=-.62, P<.0001, respectively). Receiver operating characteristic (ROC) plots for serum CC, B2M and creatinine for detection of changes in the eCcr showed that the area under the ROC curve+/-S.E. is 0.897+/ 0.119 for CC, 0.871+/-0.091 for B2M and 0.785+/-0.087 for serum creatinine. There was no statistically significant difference between the areas under the curve (AUC) for serum creatinine and CC (P=.07) and B2M (P=.12). CC had the highest sensitivity for detection of eCcr (<60 ml/min/1.73 m(2)) at routinely used cutoff values. CC was also the best discriminator when patients with normoalbuminuria were compared with patients with microalbuminuria. CONCLUSION: Although there is no significant difference in the overall diagnostic accuracies of CC, B2M and creatinine for the detection of changes in the GFR, CC is the most sensitive marker at routinely used cutoff values and would be more clinically useful than B2M or serum creatinine in diabetic patients. PMID- 12738402 TI - Natural killer cell and proinflammatory cytokine responses to mental stress: associations with heart rate and heart rate variability. AB - Associations between natural killer (NK) cell, proinflammatory cytokine stress responsivity, and cardiac autonomic responses (indexed by heart rate and heart rate variability) were assessed in 211 middle-aged men and women. Blood was drawn at baseline, immediately following color-word interference and mirror tracing tasks for the assessment of NK cell numbers, and 45 min post-stress for assessing plasma interleukin 6 (IL-6) and tumor necrosis factor alpha (TNFalpha) responses. Heart rate variability was measured as the root mean square of successive differences (RMSSD) in R-R intervals. Increases in NK cell counts following stress were positively associated with heart rate responses independently of age, sex, socioeconomic status, smoking, and change in hematocrit. Heart rate 45 min post-stress was positively associated with plasma IL-6 post-stress, and with TNFalpha changes from baseline, independently of covariates. No relationship between immune responses and heart rate variability was observed. We conclude that individual differences in sympathetically-driven cardiac stress responses are associated with NK and proinflammatory cytokine responses to psychological stress. PMID- 12738403 TI - Blind Braille readers mislocate tactile stimuli. AB - In a previous experiment, we observed that blind Braille readers produce errors when asked to identify on which finger of one hand a light tactile stimulus had occurred. With the present study, we aimed to specify the characteristics of this perceptual error in blind and sighted participants. The experiment confirmed that blind Braille readers mislocalised tactile stimuli more often than sighted controls, and that the localisation errors occurred significantly more often at the right reading hand than at the non-reading hand. Most importantly, we discovered that the reading fingers showed the smallest error frequency, but the highest rate of stimulus attribution. The dissociation of perceiving and locating tactile stimuli in the blind suggests altered tactile information processing. Neuroplasticity, changes in tactile attention mechanisms as well as the idea that blind persons may employ different strategies for tactile exploration and object localisation are discussed as possible explanations for the results obtained. PMID- 12738404 TI - Gender differences in response to pictures of nudes: a magnetoencephalographic study. AB - The magnetic equivalent of the contingent negative variation (CMV) and visual evoked magnetic fields (VEF) in anticipation of pictures of opposite-sex nude, same-sex nude, and neutral photographs has been recorded with whole-head MEG in 12 males and 12 females. Subjective ratings of valence indicated a strong gender effect. While females rated both male and female nudes as neutral, males rated male nudes similar to neutrals but female nudes received higher scores of pleasantness. Gender differences were also found for ratings of picture-induced arousal. While females rated male and female nudes as equally arousing, males attributed more arousal to opposite-sex nudes. The CMV instead revealed, for both male and female participants, higher amplitudes for opposite-sex nudes. VEF in response to nudes revealed two components with mean latencies of 126 and 203 ms. The amplitude of the first component was stronger in males than in females, and only in males the magnetic activity was increased in response to male and female nudes compared to neutral pictures. For the second component the mean magnetic activity was higher in response to nudes than to neutral contents for both male and female participants. The results are discussed in terms of an evolutionary view of sexual selection, which predicts a greater response in male subjects to stimuli relevant for mate choice. PMID- 12738406 TI - Attentional focus influences the walk-run transition in human locomotion. AB - This study tested the hypothesis that cognitive perceptual processes are involved in determining the walk-run transition in human locomotion. In a dual-task paradigm, 12 healthy male participants (aged 21.8+/-2.4 years) walked and ran on a treadmill while solving mental arithmetic (MA) organized in two levels of difficulty (easy and hard). Speed was increased over seven increments of 0.1 ms( 1) while the walk-run transition speed (TS) and central and peripheral ratings of perceived exertion (RPE) were recorded. MA performance was maintained between control (no locomotion) and treadmill conditions and participants rated hard MA as more difficult and more mentally engaging than easy MA. The TS increased during both levels of MA, although RPE values did not reflect psychological attenuation across the transition. Together, these results support the hypothesis that cognitive load distracts attentional focus from physiological cues that contribute to triggering human gait transitions. PMID- 12738405 TI - Heart rate variability during sleep as a function of the sleep cycle. AB - In this work, in order to evaluate whether autonomic differences distinguish REM sleep and NREM sleep through the whole sleeping period, statistical analysis on spectral power associated with low frequency and high frequency bands were performed on the whole polysomnographic recording, considering the sleep cycle as a unit of sleep. Our results from nine subjects show that power associated with low frequency is higher in REM sleep than in NREM sleep, while power associated with high frequency is significantly higher in NREM sleep than in REM sleep. Differences between REM sleep and NREM sleep are not of the same magnitude within the whole sleep episode and, independent of sleep stages, specific trends are observable in the autonomic control of heart rate during the night. PMID- 12738407 TI - Sensation seeking and startle modulation by physically threatening images. AB - The potential moderating effect of sensation seeking on anxious reactivity to threatening experiences was assessed using the affective modulation of startle blink paradigm. Startle blinks, as measured by electromyographic (EMG) activity in response to loud (100 dB) white-noise stimuli, were elicited during the presentation of positive, neutral, and threatening visual images. Unlike participants low in sensation seeking who showed blink potentiation during threatening versus neutral images, participants high in sensation seeking showed equal magnitudes of startle to neutral and threatening images. The results suggest that individuals high compared with low on sensation seeking are less anxiously reactive to physically threatening visual stimuli. No attenuation in startle magnitude was elicited by positive images among low or high sensation seekers suggesting that the positive images employed in the current study were not arousing enough to activate the appetitive arousal system. PMID- 12738408 TI - VIP: a very important peptide in T helper differentiation. PMID- 12738409 TI - Bioinformatics for characterisation of allergens, allergenicity and allergic crossreactivity. PMID- 12738410 TI - Beta2-microglobulin as a potential initiator of inflammatory responses. PMID- 12738412 TI - How to "spot" a real killer. PMID- 12738414 TI - Melanoma immunotherapy: a battle against radicals? PMID- 12738416 TI - Linear "2-0-1" lymphocyte development: hypotheses on cellular bases for immunity. PMID- 12738417 TI - Tolerance, DCs and tryptophan: much ado about IDO. PMID- 12738418 TI - Altering T-cell activation by targeting the multidomain tyrosine kinase Itk. PMID- 12738419 TI - Proinflammatory cytokines and sepsis syndrome: not enough, or too much of a good thing? PMID- 12738420 TI - Rewiring the T-cell: signaling defects and novel prospects for the treatment of SLE. PMID- 12738421 TI - Invariant chain, a chain of command. PMID- 12738422 TI - The immune response to parasitic helminths: insights from murine models. PMID- 12738423 TI - Viral inhibition of MHC class II antigen presentation. PMID- 12738424 TI - Is somatosensory function abnormal in amyotrophic lateral sclerosis/parkinsonism dementia complex in Kii Peninsula? PMID- 12738425 TI - Transcranial magnetic stimulation and epilepsy. AB - Epileptic conditions are characterized by an altered balance between excitatory and inhibitory influences at the cortical level. Transcranial magnetic stimulation (TMS) provides a noninvasive evaluation of separate excitatory and inhibitory functions of the cerebral cortex. In addition, repetitive TMS (rTMS) can modulate the excitability of cortical networks. We review the different ways that TMS has been used to investigate pathophysiological mechanisms and effects of antiepileptic drugs in patients with epilepsy and epileptic myoclonus. The safety of different TMS techniques is discussed too. Finally, we discuss the therapeutic prospects of rTMS in this field. PMID- 12738426 TI - Effects of auditory stimulus intensity and hearing threshold on the relationship among P300, age, and cognitive function. AB - OBJECTIVE: Although P300 is regarded as cognitive or endogenous, studies have demonstrated that stimulus intensity influences the component. To isolate effects of hearing from cognition, two experiments were designed to compare the effects of variation in stimulus intensities with naturally occurring differences in hearing thresholds. METHODS: In experiment 1, 18 participants were tested with 5 auditory oddball event-related potential (ERP) paradigms with different intensities. In experiment 2, an auditory oddball ERP task was completed by 3 groups of participants with different hearing thresholds (n=57). P300 was then correlated with block design and matrices from Wechsler's abbreviated scale of intelligence. RESULTS: At Cz and Pz, manipulation of intensity had less effect on P300 than the observed differences between groups with different hearing thresholds. At Fz, however, the effect of manipulations of stimulus intensity was greater than the effect of naturally occurring differences in hearing thresholds. P300 still correlated in predicted directions with cognitive tests after correcting for the estimated effect of differences in hearing. CONCLUSIONS: The results indicate that P300 is an index of cognitive function even when the relationship is corrected for perceptual differences, at least at posterior scalp areas. PMID- 12738428 TI - The electrophysiological net response ('F-complex') to spatial fusion of speech elements forming an auditory object. AB - OBJECTIVE: The purpose of this study was to define and analyze the brain activity associated with fusion of speech elements to form an auditory object and to study the effects of presenting the elements at different spatial locations (duplex stimulus). METHODS: Stimuli were formant transitions (presented to the front, left or right of the subject) and base (presented to the front), that fused to result in V-C-V sequences /aga/ and /ada/. Ten right-handed, adult, native Hebrew speakers discriminated each fused stimulus, and the brain potentials associated with performance of the task were recorded from 21 electrodes. The net-fusion response, the 'F(fusion)-complex', was extracted by subtracting the sum of potentials to the base and formant transitions from the potentials to the fused sound. Low resolution electromagnetic tomography analysis (LORETA) was performed to assess the timing and brain location of the fusion process. RESULTS: The 'F complex', comprising of the difference N(1), P(2), N(2b) (FN(1), FP(2), FN(2b)) components could be identified for each of the stimuli and reflected a process indicating inhibition, occlusion or both, with right ear advantage in fusion. LORETA analyses indicate sequential processing of speech fusion in the temporal lobes, beginning with right prominence in FN(1) and FP(2) shifting to a more symmetrical pattern in FN(2). CONCLUSIONS: The electrophysiological correlates of speech fusion highlight the uniqueness of speech perception and the brain areas involved in its analysis. PMID- 12738427 TI - Development of the automatic mismatch response: from frontal positivity in kindergarten children to the mismatch negativity. AB - OBJECTIVE: The automatic event-related potential (ERP) response to auditory deviance typically consists of a frontocentral mismatch negativity (MMN), which has been shown to be quite stable during development. Whereas in some infant studies, positive frontal mismatch responses have been reported instead of a MMN; to date, such positivities have not been reported for older children. METHODS: Oddball sequences with small frequency and phoneme deviance (standard: 1000 Hz, 'ba'; larger deviance: 1060 Hz, 'ta'; smaller deviance: 1030 Hz, 'da') and short intervals (every 0.38 s) were presented to 6-7-year-old children and adults during 43-channel ERP recordings. RESULTS: Children showed a consistent frontal positive mismatch response with posterior negativity (179-207 ms), and adults a frontocentral MMN with mastoid positivity (129-199 ms). This map polarity reversal was reflected by significantly different 3D centroid distributions. Low resolution electromagnetic tomography (LORETA) revealed temporal mismatch response sources for both age groups and conditions. CONCLUSIONS: Major developmental changes characterise the automatic mismatch response for the small deviances and short intervals used. Source localisation suggests that children's and adults' mismatch responses originated from superior temporal plane generators with similar localisation but opposite polarity. This indicates qualitatively different neurophysiological functioning of the automatic bi-temporal auditory change detectors in children and adults. PMID- 12738429 TI - Predicting EEG responses using MEG sources in superior temporal gyrus reveals source asynchrony in patients with schizophrenia. AB - OBJECTIVE: An integrated analysis using Electroencephalography (EEG) and magnetoencephalography (MEG) is introduced to study abnormalities in early cortical responses to auditory stimuli in schizophrenia. METHODS: Auditory responses were recorded simultaneously using EEG and MEG from 20 patients with schizophrenia and 19 control subjects. Bilateral superior temporal gyrus (STG) sources and their time courses were obtained using MEG for the 30-100 ms post stimulus interval. The MEG STG source time courses were used to predict the EEG signal at electrode Cz. RESULTS: In control subjects, the STG sources predicted the EEG Cz recording very well (97% variance explained). In schizophrenia patients, the STG sources accounted for substantially (86%) and significantly (P<0.0002) less variance. After MEG-derived STG activity was removed from the EEG Cz signal, the residual signal was dominated by 40 Hz activity, an indication that the remaining variance in EEG is probably contributed by other brain generators, rather than by random noise. CONCLUSIONS: Integrated MEG and EEG analysis can differentiate patients and controls, and suggests a basis for a well established abnormality in the cortical auditory response in schizophrenia, implicating a disorder of functional connectivity in the relationship between STG sources and other brain generators. PMID- 12738430 TI - Enhanced activation of the auditory cortex in patients with inner-ear hearing impairment: a magnetoencephalographic study. AB - OBJECTIVE: Injury of peripheral auditory organ often induces abnormality of loudness sensation such as loudness recruitment. However, objective evaluation of this phenomenon has rarely been performed. To elucidate this abnormal loudness sensation, cortical mechanisms were investigated by recording auditory evoked magnetic fields (AEFs). METHODS: We recorded AEFs in 8 patients suffering from inner-ear hearing impairment with loudness recruitment and in 14 healthy hearing controls using a 122-channel whole-head neuromagnetometer. Tone bursts of 1 kHz were presented monaurally at 4 different intensities (40, 50, 60, 70 dB HL) with a constant interstimulus interval of 1 s. RESULTS: In both groups, the 100 ms response (N100m) increased in amplitude and decreased in latency as a function of stimulus intensity in both hemispheres. Concerning the source strength, increment of dipole moment of N100m was more rapid according to the stimulus intensity in patients compared with that in healthy subjects. Source strength of N100m was enhanced at high stimulus intensity in patients, and its ratio to healthy subjects was 1.08 at 50 dB, 1.69 at 60 dB and 2.04 at 70 dB. CONCLUSIONS: In patients with inner-ear hearing impairment, enhanced activation of the auditory cortex was observed, and may help explain loudness recruitment. PMID- 12738431 TI - Transcranial magnetic stimulation (TMS) of the sensorimotor cortex and medial frontal cortex modifies human pain perception. AB - OBJECTIVE: Although recent neuroimaging studies have shown that painful stimuli can produce activity in multiple cortical areas, the question remains as to the role of each area in particular aspects of human pain perception. To solve this problem we used transcranial magnetic stimulation (TMS) as an 'interference approach' tool to test the consequence on pain perception of disrupting activity in several areas of cortex known to be activated by painful input. METHODS: Weak CO(2) laser stimuli at an intensity around the threshold for pain were given to the dorsum of the left hand in 9 normal subjects. At variable delays (50, 150, 250, 350 ms) after the onset of the laser stimulus, pairs of TMS pulses (dTMS: interpulse interval of 50 ms, and stimulus intensity of 120% resting motor threshold) were applied in separate blocks of trials over either the right sensorimotor cortex (SMI), midline occipital cortex (OCC), second somatosensory cortex (SII), or medial frontal cortex (MFC). Subjects were instructed to judge whether or not the stimulus was painful and to point to the stimulated spot on a drawing of subject's hand. RESULTS: Subjects judged that the stimulus was painful on more trials than control when dTMS was delivered over SMI at 150-200 ms after the laser stimulus; the opposite occurred when dTMS was delivered over MFC at 50 100 ms. dTMS over the SII or OCC failed to alter the pain threshold. CONCLUSIONS: These results suggest that TMS to SMI can facilitate whereas stimulation over MFC suppresses central processing of pain perception. Since there was no effect of dTMS at any of the scalp sites on the localization task, the cortical locus for point localization of pain may be different from that for perception of pain intensity or may involve a more complex mechanism than the latter. SIGNIFICANCE: This is the first report that TMS of SMI facilitates while that of MFC suppresses the central processing of pain perception. This raises the possibility of using TMS as a therapeutic device to control pain. PMID- 12738432 TI - Sleep architecture, slow wave activity and sleep spindles in mild sleep disordered breathing. AB - OBJECTIVE: A high degree of sleep fragmentation by arousals related to respiratory events would result in an abnormal distribution of slow wave activity (SWA) and a decrease in sleep spindle density in sleep disordered breathing (SDB) patients when compared to controls. METHODS: Eighteen mild SDB subjects (6 females and 12 males), aged 18-56 years with (536 h. The 95th percentile for the 2-day recordings in control subjects was 5.3%, slightly higher than observed with conventional systems. PMID- 12738451 TI - Pharmacologic therapy for the irritable bowel syndrome. AB - The management of the irritable bowel syndrome (IBS) remains unsatisfactory. For abdominal pain, antispasmodics are, at best, of only modest efficacy. Tricyclic antidepressants in low dose are useful (with the number needed to treat being three), but side effects and patient concerns regarding use of a centrally acting agent for depression remain limitations. Selective serotonin reuptake inhibitors are of uncertain efficacy in IBS. Opioid agonists, especially loperamide, are useful for diarrhea but not for pain in IBS; rebound constipation also remains a problem. Bile salt sequestering agents are not of established value in IBS but seem to be useful clinically in a small group of IBS patients with diarrhea. Aloestron, a 5HT(3) antagonist, should be reserved, if available, for women with severe diarrhea predominant IBS who have failed to respond to conventional therapy, and started at a low dose. Fiber and bulking agents may help constipation in some trials, but the evidence that they are efficacious in IBS is equivocal; they are frequently prescribed as first-line drugs for IBS regardless of the primary bowel disturbance but often increase bloating, gas, and pain. Laxatives are not of established value in IBS but are often taken by patients with constipation predominant IBS. Tegaserod, a partial 5HT(4) agonist, is now available in the United States and other countries for use in women with IBS whose primary bowel symptom is constipation; its efficacy in men and in those with alternating bowel habits is unknown. Probiotics are of uncertain efficacy. Chinese herbal medicine data are insufficient. Other new drugs in development include the cholecystokinin antagonists and novel visceral analgesics. Both current and potential therapies for IBS are reviewed in this article. PMID- 12738452 TI - Intestinal metaplasia at the gastroesophageal junction: Barrett's, bacteria, and biomarkers. AB - For patients found to have intestinal metaplasia at the gastroesophageal junction, technical problems can make it difficult to distinguish short-segment Barrett's esophagus from intestinal metaplasia of the gastric cardia. Whereas the risk of malignancy for the former condition seems to be higher than that for the latter, the distinction between these conditions can have practical clinical implications. Immunostaining for cytokeratins has been proposed as a means to distinguish intestinal metaplasia of esophageal and gastric origins. We review recent data on this issue, and conclude that immunostaining for cytokeratins has no clear advantages over other biomarkers that have been proposed for identifying Barrett's esophagus (e.g., mucin histochemistry, mAb Das-1 immunoreactivity). Presently, the importance of intestinal metaplasia at the gastroesophageal junction remains unclear, and the clinical utility of biomarkers in distinguishing short-segment Barrett's esophagus from intestinal metaplasia of the gastric cardia has not yet been established. PMID- 12738453 TI - Costs and clinical outcomes of primary prophylaxis of variceal bleeding in patients with hepatic cirrhosis: a decision analytic model. AB - OBJECTIVE: Current guidelines recommend upper endoscopic screening for patients with hepatic cirrhosis and primary prophylaxis with a nonselective beta-blocker for those with large varices. METHODS: However, only 25% of cirrhotics develop large varices. Thus, the aim of this study is to evaluate the most cost-effective approach for primary prophylaxis of variceal hemorrhage. RESULTS: Using a Markov model, we compared the costs and clinical outcomes of three strategies for primary prophylaxis of variceal bleeding. In the first strategy, patients were given a beta-blocker without undergoing upper endoscopy. In the second strategy, patients underwent upper endoscopic screening; those found to have large varices were treated with a beta-blocker. In the third strategy, no prophylaxis was used. Selected sensitivity analyses were performed to validate outcomes. Our results show screening prophylaxis was associated with a cost of $37,300 and 5.72 quality adjusted life yr (QALYs). Universal prophylaxis was associated with a cost of $34,100 and 6.65 QALYs. The no prophylaxis strategy was associated with a cost of $36,600 and 4.84 QALYs. The incremental cost-effectiveness ratio was $800/QALY for the endoscopic strategy relative to the no prophylaxis strategy. Screening endoscopy was cost saving when the compliance, bleed risk without beta-blocker, and variceal bleed costs were increased, and when the discount rate, bleed risk on beta-blockers, and cost of upper endoscopy were decreased. In contrast, the universal prophylaxis strategy was persistently cost saving relative to the no prophylaxis strategy. In comparing the strategies, sensitivity analysis on the death rates from variceal hemorrhage did not alter outcomes. CONCLUSIONS: Our results provide economic and clinical support for primary prophylaxis of esophageal variceal bleeding in patients with hepatic cirrhosis. Universal prophylaxis with beta-blocker is preferred because it is consistently associated with the lowest costs and highest QALYs. PMID- 12738454 TI - Ineffective motility is not a marker for gastroesophageal reflux disease. AB - OBJECTIVE: Previous studies have suggested that ineffective esophageal motility (IEM) may be a marker for gastroesophageal reflux disease (GERD), particularly supraesophageal reflux disease. We evaluated the relationship between esophageal acid exposure and esophageal body motility in patients undergoing both esophageal manometry and 24-h pH metry in the absence of antisecretory therapy. METHODS: We conducted a retrospective database review of 84 patients (mean age 47 yr, 46% male) evaluated in our GI physiology laboratory. The indication for testing was recorded and characterized as esophageal or supraesophageal. Abnormal esophageal acid exposure was defined as a distal esophageal pH <4 for more than 4.2% of the total monitoring time (>6.3% upright, >1.2% supine) or a proximal esophageal acid exposure time of greater than 1.1% total (>1.3% upright, 0% supine). IEM was defined as more than two of 10 ineffective peristaltic waves. RESULTS: Seventy two patients had esophageal-presenting symptoms, and 12 had supraesophageal symptoms. The prevalence of abnormal esophageal acid exposure was similar in patients with esophageal and supraesophageal symptoms (69% vs 92%, p = 0.17). Abnormal motility was identified in 26 patients (31%). IEM was the most common motility disturbance (77%, 20 patients). The frequency of motility disorders was similar in patients with and without abnormal esophageal acid exposure (30% vs 35%, p = 0.79), in patients with esophageal or supraesophageal symptoms (32% vs 25%, p = 0.75, for all patients; 30% vs 27%, p = 1.00, for patients with abnormal esophageal acid exposure), and among upright, supine, and combined refluxers (33%, 9%, and 35%, p = 0.26). CONCLUSIONS: IEM does not stand alone as a significant marker for the presence of GERD in general or supraesophageal reflux disease in particular. PMID- 12738455 TI - Elemental diet is an effective treatment for eosinophilic esophagitis in children and adolescents. AB - OBJECTIVE: Eosinophilic esophagitis (EoE), a disorder characterized by eosinophilic infiltration of the esophageal mucosa, has been defined in large part through published case reports and series leading to ambiguity in both diagnostic and treatment options. Corticosteroids, cromolyn, and elemental diet have all been reported as successful treatments for EoE. In this study, we sought to accurately define a population of patients with EoE and then assess their response to elemental diet. METHODS: A series of patients with chronic symptoms of gastroesophageal reflux disease and an isolated esophageal eosinophilia on esophagogastroduodenoscopy (EGD) were identified. Therapy with a proton pump inhibitor was instituted for 3 months, followed by repeat EGD when symptoms persisted. A 24-h pH probe study was performed, and those with significantly abnormal studies were excluded. The remaining patients were diagnosed with EoE and placed on an elemental diet for 1 month, followed by a repeat EGD. RESULTS: Of 346 patients with chronic gastroesophageal reflux disease symptoms and eosinophils on esophageal biopsy, 51 (14.7%) were ultimately diagnosed with EoE. There was significant improvement in vomiting, abdominal pain, and dysphagia after the elemental diet. The median number of esophageal eosinophils per high powered field (HPF) decreased from 33.7 before the diet to 1.0 after the diet (p <0.01). The average time to clinical improvement was 8.5 days. CONCLUSIONS: Elemental diet resulted in striking improvement in both symptoms and histologic evidence of disease in children and adolescents with EoE, as identified by strict diagnostic criteria. PMID- 12738456 TI - Symptoms associated with impaired gastric emptying of solids and liquids in functional dyspepsia. AB - OBJECTIVE: The relationship between functional dyspepsia and delayed gastric emptying of solids or liquids is still unclear. The aim of the present study was to investigate in dyspeptic patients the prevalence of delayed gastric emptying for solids or for liquids and to investigate the relationship to the dyspepsia symptom pattern. METHODS: In 392 and 330 patients with functional dyspepsia, the solid and liquid gastric emptying, respectively, was measured using breath tests, and the severity of eight dyspeptic symptoms was scored. RESULTS: Gastric emptying of solids and liquids were delayed in 23% and 35% of the patients. Multivariate analysis showed that the presence of vomiting and postprandial fullness was associated with delayed solid emptying (OR 2.65, 95% CI = 1.62-4.35 and OR 3.08, 95% CI = 1.28-9.16, respectively). Postprandial fullness was also associated with the risk of delayed liquid emptying when symptom was present (OR 3.5, 95% CI = 1.57-8.68), relevant or severe (OR 2.504, 95% CI = 1.41-4.65), and severe (OR 2.214, 95% CI = 1.34-3.67). Severe early satiety was associated with the risk of delayed liquid emptying (OR 1.902, 95% CI = 1.90-3.30). CONCLUSIONS: A subset of dyspeptic patients has delayed gastric emptying of solids or of liquids. Delayed gastric emptying of solids was constantly associated with postprandial fullness and with vomiting. Delayed emptying for liquids was also associated with postprandial fullness and with severe early satiety. PMID- 12738457 TI - Does psychological distress modulate functional gastrointestinal symptoms and health care seeking? A prospective, community Cohort study. AB - OBJECTIVE: Little is known about the natural history of functional GI symptoms, including what factors influence GI symptom patterns and health care seeking for them over the long term. We aimed to determine whether psychological factors play a role in the development and long-term course of these symptoms. METHODS: A random sample of community subjects (n = 361) who reported having unexplained abdominal pain for > or =1 month in a previous population survey were included in the study. Controls (n = 120) were defined as not having abdominal pain for > or =1 month from this initial survey. Subjects were prospectively followed up via a questionnaire every 4 months over a 12-month period. The questionnaire asked about the presence of GI symptoms over the past week and psychological distress over the past 3 wk (psychological caseness being defined as a score of > or=2 of 12 on the General Health Questionnaire). The number of visits made to a physician or medical specialist over the past 4 months was evaluated. RESULTS: GI symptoms were common among community controls and subjects with abdominal pain. Similar onset and disappearance rates were observed for the majority of GI symptom categories, accounting for the stability of the prevalence rates over a 1-yr period. Changes in a state measure of psychological distress were not significantly associated with changes in GI symptom status between the 4- and 8 month (r = 0.14, p = 0.08) and 8- and 12-month (r = 0.02, p = 0.77) follow-ups. Baseline psychological distress, however, was an independent predictor of having persistent GI symptoms, including abdominal pain, bloating, and constipation, and frequently seeking health care for GI symptoms over 1 yr. CONCLUSIONS: Psychological distress levels do not seem to be important in explaining GI symptom change over a 1-yr period. Psychological distress, however, is linked to having persistent GI symptoms and frequently seeking health care for them over time. Clinicians should consider psychological factors in the treatment of this subset of irritable bowel syndrome patients. PMID- 12738458 TI - Lack of accuracy of the noninvasive Helicobacter pylori stool antigen test in patients with gastroduodenal ulcer bleeding. AB - OBJECTIVE: Oral presentation at Digestive Diseases Week, San Francisco, California, May 2002. The antigen-based stool assay has proven to be accurate in diagnosing Helicobacter pylori infection in dyspeptic patients. We evaluated the H. pylori antigen-based stool assay (HpSA) in patients with peptic ulcer bleeding (PUB). METHODS: Thirty-six patients with PUB were endoscoped, and antral and corpus biopsy specimens were taken for rapid urease test (RUT), histology, and culture. The first stool sample after admission was collected for the HpSA test. The gold standard was defined as either positive culture or positive RUT and histology. If only RUT or histology was positive, this was defined as indeterminate. To evaluate cross-reaction with blood constituents, citrated blood samples from 10 healthy volunteers (nine H. pylori serology negative and one H. pylori serology positive) were assessed by the HpSA test. RESULTS: A total of 36 consecutive patients with PUB (21 male) with a mean age of 69.5 yr were included in the study. Using the gold standard, the sensitivity and specificity of the HpSA test were 100% and 52%, respectively. Citrated blood samples of three H. pylori negative and one H. pylori positive volunteer gave a positive result in the HpSA test, suggesting cross-reaction with blood con stituents. CONCLUSIONS: The HpSA test gave a high number of false- positive results in patients with PUB, probably because of blood constituents cross-reacting in the enzyme immunoassay. The HpSA test is not accurate for testing H. pylori infection in patients with PUB. PMID- 12738459 TI - Endoscopic findings of diverticular inflammation in colonoscopy patients without clinical acute diverticulitis: prevalence and endoscopic spectrum. AB - OBJECTIVE: Experienced colonoscopists sometimes encounter endoscopic findings of diverticular inflammation in patients without clinical evidence of acute diverticulitis. Our aim was to describe the spectrum and prevalence of such endoscopic findings in a consecutive series of patients undergoing colonoscopy. METHODS: During elective colonoscopy performed by a single endoscopist on 2566 consecutive outpatients, 21 patients were identified with endoscopic evidence of diverticular inflammation. RESULTS: Endoscopic findings included erythema and edema of a diverticular opening (n = 8), pus emanating from a diverticular orifice (n = 8), and a polypoid mass of granulation tissue in a diverticular orifice (n = 15). Follow-up was obtained by telephone in 17 patients at a mean of 11.9 months after colonoscopy. Only one patient had symptoms of diverticulitis at the time of colonoscopy. This patient improved with antibiotic therapy. Six of the 21 patients had experienced symptoms of abdominal pain or fever, but only one had a diagnosed episode of acute diverticulitis before colonoscopy. None of the patients had acute diverticulitis during the follow-up interval. CONCLUSIONS: Endoscopic findings of diverticular inflammation were identified in about 0.8% of patients undergoing colonoscopy without clinical evidence of diverticulitis. Most patients are asymptomatic at the time of colonoscopy, and antibiotic therapy is generally unnecessary. PMID- 12738460 TI - Prospective evaluation of a 3.1-mm battery-powered esophagoscope in screening for esophageal varices in cirrhotic patients. AB - OBJECTIVE: Standard esophagogastroduodenoscopy (EGD) is costly and uses conscious sedation that cirrhotic patients may tolerate poorly. This study aimed to determine the feasibility and acceptance of unsedated esophagoscopy with an ultrathin battery-powered endoscope (BPE) in cirrhotic patients for diagnosing esophageal varices (EV). METHODS: We first studied the prevalence of significant gastroduodenal pathology that could be missed if only esophagoscopy were performed in cirrhotic patients undergoing liver transplant evaluation. A prospective study was then done to evaluate a BPE in EV screening. Unsedated per oral endoscopy was first done by a single endoscopist using a BPE, followed by EGD by a second endoscopist who was masked to the BPE result. A visual analog score was used to determine patient tolerance. Patients were asked about their preference for endoscopy in the future. A paired Student t test and the kappa statistic were used in the statistical analysis. RESULTS: In the retrospective study, 199 patients were reviewed; three patients (1.5%) had gastric ulcers, and two patients (1%) had duodenal ulcers. In the prospective study, 28 cirrhotic patients (16 women) were evaluated. EV were diagnosed in 14 patients with a BPE, and 13 were confirmed by standard EGD (sensitivity and negative predictive value 100%, specificity and positive predictive value 93%, kappa = 0.93). EV were graded as large in one and small in 13 patients with a BPE, but small varices diagnosed in one patient were not confirmed on EGD. Both procedures were well tolerated by all patients. Twenty-seven of 28 patients preferred unsedated endoscopy with a BPE over EGD. CONCLUSIONS: Unsedated endoscopy with a BPE is safe and well tolerated. The diagnostic accuracy of a BPE for diagnosing EV is the same as by EGD. Esophagoscopy with a BPE is a potential alternative to EGD for EV screening. PMID- 12738461 TI - Incidence of gallstones in chronic renal failure patients undergoing hemodialysis: experience of a center in Turkey. AB - OBJECTIVE: In this case-control study, we sought to determine whether the incidence of gallbladder stones (GBS) was increased in chronic renal failure (CRF) patients on a hemodialysis (HD) program. We also evaluated factors, such as lipid profiles and gallbladder motility, that could affect the formation of GBS. In addition, we reviewed other available studies on this subject and compared the factors that might have some influence on the development of GBS. METHODS: A total of 182 CRF patients (135 male, 47 female, mean age 32.1 yr) undergoing chronic HD and who were referred to our transplantation center in the last 10 yr and 194 healthy controls (137 male, 57 female, mean age 33.3 yr) were included in the study. Abdominal ultrasound was performed on all patients, and ALT, AST, and lipid profiles were determined. In addition, 19 patients with CRF (12 male, 7 female, mean age 33.5 yr) and 22 controls (14 male, 8 female, mean age 33.2 yr) who were age and sex matched were randomly chosen for gallbladder emptying, monitored by ultrasound at 30-min intervals for 2 h after a mixed meal. Fasting volume, minimal residual volume, and ejection fraction of the gallbladder were assessed. For statistical analysis, chi(2), t test, and logistic regression analysis were used. RESULTS: GBS were detected in seven patients with CRF (3.85%, 5 male, 2 female) and three controls (1.55%, one male, two female) (p > 0.05). The mean follow-up time of CRF patients after diagnosis was 39.3 months (range: 2 168), the mean duration of HD was 21.8 months (range: 1-120). The analysis of seven stones in the CRF group revealed that five were cholesterol-rich stones, and two were mixed (cholesterol and bilirubin) stones. Cholesterol levels were higher in the control group, and triglycerides were higher in the CRF group, but these findings were nonsignificant (p > 0.05). Other biochemical values were not significantly different between the groups. CRF patients with and without GBS were similar in their duration of CRF and HD, age, and other biochemical parameters (p > 0.05). When gallbladder emptying was considered, there was no difference between the two groups in fasting volume, residual volume, and ejection fraction (CRF: 89.7%; controls: 92.3%) of the gallbladders (p > 0.05). CONCLUSIONS: We detected similar incidences of GBS in CRF patients undergoing HD and healthy controls, and this was comparable to the results of most of the previous studies. Young male CRF patients had a nonsignificantly higher incidence of GBS than control males. Although cholesterol-rich GBS were predominant, we could not find any significant difference between the groups when factors that could affect GBS formation, such as lipid profiles and gallbladder motility, were taken into account. PMID- 12738462 TI - Endoscopic stent therapy in advanced chronic pancreatitis: relationships between ductal changes, clinical response, and stent patency. AB - OBJECTIVE: Pancreatic duct stenting is now recognized as a treatment option for a number of pancreatic disorders. Although stent-induced ductal changes may result, there is little information regarding the frequency of these stent-induced changes in chronic pancreatitis and their relationship to stent occlusion and clinical response. Our objectives were to evaluate pancreatic ductal changes after endoscopic stenting in patients with preexisting radiographic evidence of chronic pancreatitis and to evaluate the relationships between ductal changes, pain response, and stent patency. METHODS: Twenty-five consecutive patients had 40 stent placement episodes. Main pancreatic duct diameter, pancreatitis grade, preexisting obstructive lesions, and stent-induced strictures were recorded. Pain response and stent patency were correlated with main pancreatic duct caliber change using chi(2) analysis. RESULTS: In 28 (70%) of 40 episodes, main pancreatic duct caliber increased or was unchanged after stenting; pain improved in 20 (71%) of 28. Pain improved in six (50%) of 12 patients with smaller ducts after stenting. Stent patency was documented upon retrieval in 34 episodes; most stents were occluded. Stent-induced strictures developed in 18% of 40 stent episodes. CONCLUSIONS: Main pancreatic duct caliber after endoscopic stenting was not a good indicator of pain response or stent patency; main pancreatic duct was often larger, and even with stent occlusion, patients' symptoms were frequently improved. Stent-induced strictures were infrequent, compared with values previously reported in the literature. PMID- 12738463 TI - Low volume bowel preparation for colonoscopy: randomized, endoscopist-blinded trial of liquid sodium phosphate versus tablet sodium phosphate. AB - OBJECTIVE: The aim of this study was to compare the colon-cleansing effectiveness, ease of consumption, and side effect profiles of two commercially available preparations of sodium phosphate: liquid Fleet Phospho-soda and Visicol tablets. METHODS: Outpatients undergoing elective colonoscopy were sequentially randomized to one of two preparation groups: liquid: 45 ml at 7:00 PM, 45 ml 3 h before colonoscopy; or tablet: 20 tablets at 7:00 PM, 20 tablets 3-5 h before colonoscopy. Subjects rated preparation tolerability on a 5-point Likert scale. A Residual Stool Score was calculated for each subject based on the amount of stool, consistency of residual stool, and percent of bowel visualized (range 0 11, 0 = best). The endoscopists were blinded to the preparation used. RESULTS: A total of 101 subjects were enrolled (43 male, 58 female, mean age 58.2 yr). The groups were similar in age and indications for colonoscopy. Overall, bowel cleansing was rated "Excellent" or "Good" in 92% of liquid preparation subjects, compared with 74% of tablet preparation subjects (p = 0.03). Subjects in the liquid group demonstrated significantly less residual stool than did tablet subjects (Residual Stool Scores: liquid 1.3 +/- 1.2 vs tablet 1.9 +/- 1.5, p < 0.05). Subjects rated the liquid preparation easier to swallow (p < 0.005) and more convenient to take (p < 0.005) than tablets. Among liquid subjects, 45 of 50 reported a willingness to take their preparation for future colonoscopies, compared with 36 of 49 who took tablet sodium phosphate (p < 0.04). CONCLUSIONS: Liquid sodium phosphate is better tolerated and more effective at colon cleansing when compared with sodium phosphate in tablet form. PMID- 12738464 TI - Infliximab (REMICADE) therapy in the treatment of pediatric Crohn's disease. AB - OBJECTIVE: The aim of this study was to assess the efficacy and safety of a single infusion of infliximab in the treatment of pediatric Crohn's disease (CD). METHODS: A total of 21 pediatric CD patients were enrolled at seven study centers and randomized to receive a single infusion of infliximab 1 mg/kg (n = 6), 5 mg/kg (n = 7), or 10 mg/kg (n = 8) over at least 2 hrs at week 0 in this multicenter, open-label, dose-blinded trial. Efficacy assessments, including the Pediatric Crohn's Disease Activity Index (PCDAI), modified CDAI, C-reactive protein concentration (CRP), and erythrocyte sedimentation rate (ESR) determinations, were made at screening and at weeks 1, 2, 4, 8, and 12. Adverse events were assessed throughout study participation. RESULTS: Improvements in the PCDAI, modified CDAI, ESR, and CRP were observed with all infliximab doses, beginning at week 1. On average, all treated patients experienced approximately 50% improvement in the PCDAI by week 2. By week 12, the PCDAI remained approximately 30% improved from baseline. During the study, all 21 patients (100%) achieved a clinical response, and 10 patients (48%) achieved clinical remission. There were no infusion reactions in any of the treatment arms. CONCLUSIONS: The results of this trial suggest that infliximab may be safe and effective as short-term therapy of medically refractory moderate to severe CD in a pediatric population. PMID- 12738465 TI - High prevalence of small intestinal bacterial overgrowth in celiac patients with persistence of gastrointestinal symptoms after gluten withdrawal. AB - OBJECTIVE: Celiac disease is a gluten-sensitive enteropathy with a broad spectrum of clinical manifestation, and most celiac patients respond to a gluten-free diet (GFD). However, in some rare cases celiacs continue to experience GI symptoms after GFD, despite optimal adherence to diet. The aim of our study was to evaluate the causes of persistence of GI symptoms in a series of consecutive celiac patients fully compliant to GFD. METHODS: We studied 15 celiac patients (five men, 10 women, mean age 36.5 yr, range 24-59 yr) who continued to experience GI symptoms after at least 6-8 months of GFD (even if of less severity). Antigliadin antibody (AGA) test, antiendomysial antibody (EMA) test, and sorbitol H2-breath test (H2-BT), as well as esophagogastroduodenoscopy (EGD) with histological evaluation, were performed before starting GFD. Bioptic samples were obtained from the second duodenal portion during EGD, and histopathology was expressed according to the Marsh classification. To investigate the causes of persistence of GI symptoms in these patients, we performed AGA and EMA tests, stool examination, EGD with histological examination of small bowel mucosa, and sorbitol-, lactose-, and lactulose H2-breath tests. RESULTS: Histology improved in all patients after 6-8 months of GFD; therefore, refractory celiac disease could be excluded. One patient with Marsh II lesions was fully compliant to his diet but had mistakenly taken an antibiotic containing gluten. Two patients showed lactose malabsorption, one patient showed Giardia lamblia and one patient Ascaris lumbricoides infestation, and 10 patients showed small intestinal bacterial overgrowth (SIBO) by lactulose H2-BT. We prescribed a diet without milk or fresh milk-derived foods to the patient with lactose malabsorption; we treated the patients with parasite infestation with mebendazole 500 mg/day for 3 days for 2 consecutive wk; and we treated the patients with SIBO with rifaximin 800 mg/day for 1 wk. The patients were re-evaluated 1 month after the end of drug treatment (or after starting lactose-free diet); at this visit all patients were symptom free. CONCLUSIONS: This study showed that SIBO affects most celiacs with persistence of GI symptoms after gluten withdrawal. PMID- 12738466 TI - Work losses related to inflammatory bowel disease in Canada: results from a National Population Health Survey. AB - OBJECTIVE: Few studies have assessed the influence of inflammatory bowel disease (IBD) on work loss or estimated the costs related to work loss. Our analysis reports the employment effects related to IBD as based on the 1998 sample of the Canadian National Population Health Survey (NPHS). METHODS: Our predictive analysis adapts the theory of labor supply to a health context. Respondents between the ages of 20 and 64 who reported that they had been diagnosed by a health professional to have "a bowel disorder such as Crohn's disease or colitis" were distinguished from the other respondents. A logistic regression model was used to estimate the OR for labor force nonparticipation and variables predictive of it in the case of IBD. For those people in the labor force, the Cox proportional hazard model was used to determine whether having IBD and similar bowel disorders had an effect on the number of months of continuous employment. RESULTS: Of the IBD patients 28.9% reported labor force nonparticipation, which was a greater proportion than the non-IBD respondents (18.5% nonparticipation). The OR was 1.20 (95% CI = 1.19-1.21) for nonparticipation of IBD patients versus non-IBD patients controlling for potentially confounding factors. We estimated the excess nonparticipation attributable to IBD and similar bowel disorders in Canada to be 2.9%. Based on this, the indirect cost of nonparticipation attributable to IBD in 1998 was >$104.2 million Canadian dollars. According to the second regression using the Cox proportional hazard model, IBD and similar bowel disorders were not significantly related to the number of months worked until a break in employment was reported. Thus, there was no excess work loss among those who were employed that was associated with IBD. CONCLUSIONS: By using directly observed data in our analysis, this method of estimation can predict the overall burden of IBD and similar bowel disorders, controlling for the effect of other potentially influential characteristics. PMID- 12738467 TI - Cytokine genotyping (TNF and IL-10) in patients with celiac disease and selective IgA deficiency. AB - OBJECTIVE: Selective IgA deficiency (IgAD) and celiac disease (CD) are frequently associated and share the ancestral haplotype human leukocyte antigen (HLA)-8.1, which is characterized by a peculiar cytokine profile. The aim of this study was to evaluate the role of tumor necrosis factor (TNF) and interleukin (IL)-10 alleles in CD and CD-IgAD. METHODS: The distribution of some biallelic polymorphisms of both cytokine promoters (-308G-->A and -863C-->A at TNF promoter sequence and -1082G-->A, -819C-->A, and -592C-->T at IL-10 promoter) were typed using biotilinated specific probes in 32 celiac patients, in 34 CD-IgAD patients, and in 96 healthy controls. RESULTS: In CD and CD-IgAD, the -308A allele was significantly more frequent than in controls, whereas no significant differences were observed for the biallelic polymorphisms at the -863 and for the three IL-10 promoter polymorphisms. The evaluation of combined TNF and IL-10 genotypes showed in CD-IgAD a significant reduction of -308G/-1082G homozygous subjects and both in CD and CD-IgAD groups an increase of 308AA/1082GG. Accordingly, CD-IgAD patients positive both for -308A TNF and -1082A IL-10 showed an increase of TNF alpha and a reduction of IL-10 serum levels. CONCLUSIONS: Genetically determined increased production of TNF-alpha and reduction of IL-10 may be relevant for susceptibility to CD, mainly in IgAD, as the different allele expression at TNF and IL-10 loci seems to influence cytokine production profile. PMID- 12738468 TI - Constipation, laxative use, and colon cancer in a North Carolina population. AB - OBJECTIVE: The aim of this study was to determine whether bowel movement frequency and laxative use and type were associated with risk of colon cancer in white and black men and women. METHODS: We conducted a population-based, case control study with equal representation by blacks. Eligible subjects between ages 40 and 80 yr residing in urban and rural communities in North Carolina were asked about bowel habits and laxatives during face-to-face interviews. There were 643 cases (349 white, 294 black) and 1048 controls (611 white, 437 black). RESULTS: Constipation, defined as fewer than three reported bowel movements per wk, was associated with a greater than two-fold risk of colon cancer (OR 2.36; 95% CI = 1.41-3.93) adjusted for age, race, sex, and relevant confounders. The association was greater for women (OR 2.69; 95% CI = 1.46-4.94) than for men (OR 1.73; 95% CI = 0.61-4.88) and stronger in blacks than whites. Black women had the highest risk (OR 3.42; 95% CI = 1.60-7.34), which remained significant (OR 3.21; 95% CI = 1.46 7.04) even after excluding subjects with late stage (distant) disease. The OR for constipation was slightly higher for distal than for proximal colon cancers. There was no association with laxative use (OR 0.88; 95% CI = 0.69-1.11). Fiber commercial laxatives appeared to exert a protective effect in a small subgroup. CONCLUSIONS: This study provides support for a positive association between constipation and increased risk for colon cancer. Women, especially black women with constipation, seem to be at the highest risk. PMID- 12738469 TI - Effects of nonpathogenic bacteria on cytokine secretion by human intestinal mucosa. AB - OBJECTIVE: The human intestine harbors a complex microbial ecosystem, and the mucosa is the interface between the immune system and the luminal environment. The aim of this study was to elucidate whether host-bacteria interactions influence mucosal cytokine production. METHODS: Macroscopically normal colonic specimens were obtained at surgery from eight patients with neoplasm, and inflamed ileal specimens were obtained from two patients with Crohn's disease. Mucosal explants were cultured for 24 h with either nonpathogenic Escherichia coli ECOR-26, Lactobacillus casei DN-114 001, L. casei DN-114 056, L. casei ATCC 334, or Lactobacillus bulgaricus LB-10. Each study included blank wells with no bacteria. Tissue and bacteria viability were confirmed by LDH release and culture. Concentration of tumor necrosis factor (TNF)alpha, transforming growth factor beta1, interleukin (IL)-8, and IL-10 was measured in supernatants. In parallel experiments, neutralizing anti-TNFalpha antibody was added to the culture. RESULTS: Co-culture of mucosa with bacteria did not modify LDH release. Co-culture with L. casei strains significantly reduced TNFalpha release, whereas E. coli increased it. These effects were observed both in normal and inflamed mucosa. In combination studies, L. casei DN-114 001 prevented TNFalpha stimulation by E. coli. L. casei DN-114 001 also reduced IL-8 release via a TNFalpha-independent pathway. L. casei DN-114 056 or E. coli increased IL-10 release in the presence of neutralizing anti-TNFalpha. CONCLUSIONS: Nonpathogenic bacteria interact with human intestinal mucosa and can induce changes in cytokine production that are strain specific. PMID- 12738470 TI - Abnormalities in gastrointestinal motility are associated with diseases of oxidative phosphorylation in children. AB - OBJECTIVE: Disorders of the mitochondrial electron transport chain enzymes of oxidative phosphorylation (OXPHOS) have neurologic, musculoskeletal, ophthalmologic, cardiac, and GI manifestations. Many adult and pediatric patients with disorders of OXPHOS have abnormalities in intestinal motility. The purpose of this study was to describe pediatric patients who initially presented with signs of GI dysmotility and were later evaluated and found to have a disorder of OXPHOS. METHODS: Data were collected on six patients, including initial GI and neurologic symptoms, histology of skeletal muscle biopsies, mitochondrial DNA mutational analysis, OXPHOS enzyme assay, upper GI barium imaging, technetium-99M liquid gastric emptying scan, upper GI endoscopy, esophageal manometry, and antroduodenal manometry. RESULTS: All six children presented with symptoms of GI dysmotility within 2 wk of life. Patients later developed symptoms of neurologic disorders. All patients had abnormalities in OXPHOS enzyme analysis. Muscle histology showed nonspecific changes with no ragged red fibers. Sequencing of the mitochondrial DNA showed no recognized mutations. No patient had any evidence of intestinal obstruction or malrotation by upper GI barium imaging. Four patients had delayed gastric emptying. Three patients had endoscopic and histologic evidence of esophagitis. All six had demonstrable neuropathic abnormalities by antroduodenal manometry, including the following: nonpropagated antral bursts, absent migrating motor complexes, postprandial antral hypomotility, retrograde migrating motor complexes, and tonic contractions with the migrating motor complex. CONCLUSIONS: Abnormalities in GI motility may be an early presenting sign of disorders of OXPHOS in children. PMID- 12738471 TI - Intrafamilial transmission of hepatitis C virus in patients with hepatitis C and human immunodeficiency virus coinfection. AB - OBJECTIVE: The aim of this study was to determine whether hepatitis C virus (HCV)/HIV coinfection of index cases increases intrafamilial transmission (sexual and nonsexual contacts) of HCV. METHODS: We prospectively enrolled 347 subjects, including 87 family members of 53 HCV/HIV-coinfected index cases and 134 family members of 73 HCV-monoinfected index cases, which served as a control group. All index cases and family members were interviewed, and a screening for HCV and HIV using enzyme-linked immunosorbent assays was performed. Positive samples were confirmed by polymerase chain reaction and tested for genotype and HCV RNA viral load. A meta-analysis designed to assess the pooled risk of sexual transmission of HCV among HCV/HIV-coinfected patients was performed. RESULTS: Anti-HCV was detected in 2.2% of family members of HCV-monoinfected index cases and 2.3% of family members of HCV/HIV-coinfected index cases. Viral load was higher in coinfected index cases (7.2 x 10(6) mEq/ml) compared with HCV alone (1.9 x 10(6) mEq/ml), p = 0.01. HCV genotype concordance was observed in three family members of HCV-monoinfected index cases and in two family members of HCV/HIV-coinfected index cases. The pooled OR of the meta-analysis evaluating HIV as a cofactor of sexual transmission of HCV was 1.54 (95% CI = 0.76-3.12). CONCLUSIONS: Our data demonstrate a low prevalence of intrafamilial transmission of HCV, independent of the presence of HCV/HIV coinfection. This finding is supported by meta-analysis, which failed to identify HIV as an important cofactor of sexual transmission in HCV/HIV-coinfected patients. PMID- 12738472 TI - Prevalence and incidence of cryoglobulins in hepatitis C virus-related chronic hepatitis patients: a prospective study. AB - OBJECTIVE: A high prevalence of cryoglobulins has been reported in patients with hepatitis C virus (HCV)-related liver disease. The aim of this study was to evaluate the prevalence and the incidence of cryoglobulins and their association with clinical symptoms in chronic hepatitis and cirrhosis patients. METHODS: The prevalence of cryoglobulins and cryoglobulinemic syndrome was investigated at enrollment in 237 patients (213 with chronic hepatitis and 24 with cirrhosis). A 7-yr follow-up was conducted evaluating the occurrence of cryoglobulins and/or cryoglobulinemic syndrome every 6 months. Rheumatoid factor was also tested in all patients. RESULTS: Prevalence of rheumatoid factor, cryoglobulins, and cryoglobulinemic syndrome in chronic hepatitis patients were 2%, 0.8%, and 0%, respectively. In cirrhosis patients the prevalence was 4%, 8%, and 0%, respectively. No statistically significant differences were found between the two groups. During the follow-up only one patient for each group abruptly developed cryoglobulinemic syndrome, and none of the patients who showed signs of cryoglobulinemia developed the syndrome or showed signs of evolution of the disease. CONCLUSIONS: Our data demonstrate that the presence of cryoglobulins and/or cryoglobulinemic syndrome in HCV-related liver disease is unusual, as is the occurrence of cryoglobulinemia over time in these patients. This leads us to think that HCV-related cryoglobulinemic syndrome and HCV-related liver disease are independent diseases. This supports new and indirect evidence for an independent and direct role of HCV in liver and blood disorders. PMID- 12738473 TI - Safety and efficacy of estrogen therapy in preventing bone loss in primary biliary cirrhosis. AB - OBJECTIVE: Estrogen therapy has been found to be useful in the treatment of postmenopausal osteoporosis. However, concern about its potential for worsening cholestasis has limited its use in postmenopausal women with primary biliary cirrhosis. The aim of the present study was to determine the safety as well as the efficacy of estrogen replacement therapy with respect to bone mass in postmenopausal women with primary biliary cirrhosis. METHODS: Bone mineral density of the lumbar spine (measured using dual energy x-ray absorptiometry) of 46 unselected postmenopausal women with primary biliary cirrhosis receiving estrogen replacement therapy was compared with 46 age-matched women with primary biliary cirrhosis not receiving estrogen replacement therapy, and with the expected normal bone mineral density adjusted for age and ethnic group. RESULTS: The mean duration of follow-up was 4.8 +/- 0.4 yr. Treatment with estrogens resulted in a significantly lower rate of bone loss (0.002 g/cm(2)/yr +/- 0.028 vs 0.009 g/cm(2)/yr +/- 0.020, p = 0.05). Worsening cholestasis attributable to estrogen replacement therapy did not occur in any patient. One patient (2%) discontinued therapy because of side effects. CONCLUSIONS: Estrogen replacement therapy in postmenopausal patients with primary biliary cirrhosis is safe and may be effective in decreasing the rate of bone loss. PMID- 12738474 TI - Liver assessment and biopsy in patients with marked coagulopathy: value of mini laparoscopy and control of bleeding. AB - OBJECTIVE: Evaluation of liver disease in patients with a high risk of postbiopsy bleeding presents a diagnostic challenge. Mini-laparoscopy offers the possibility of coagulation of biopsy site and the additional advantage of macroscopic liver assessment. We wished to assess the value and safety of mini-laparoscopy with guided biopsy as a diagnostic approach in patients in whom percutaneous liver biopsy is considered contraindicated because of a marked coagulopathy. METHODS: We investigated 61 consecutive patients with marked coagulopathy (prolonged international normalized ratio > 1.5, thrombocytopenia < 50/nl, or both; von Willebrand's disease/hemophilia). Diagnostic mini-laparoscopy with visually guided liver biopsy was undertaken for the evaluation of liver disease. Biopsy sites were coagulated prophylactically (n = 4) or therapeutically (n = 52). Safety, diagnostic yield, and therapeutic consequences were assessed. RESULTS: Macroscopic assessment of the liver was possible in 60/61 high-risk patients and was considered diagnostic in 1/61. In 58 of the remaining 60 patients, liver biopsy was technically feasible. There was no persistent postbiopsy bleeding. One patient with fulminant hepatic failure had self-limiting bleeding from the abdominal wall. Ninety-seven percent of the biopsies were of adequate size for diagnostic histological evaluation. CONCLUSIONS: Mini-laparoscopy with guided liver biopsy allows reliable and safe evaluation of liver disease in patients with severe coagulopathy. PMID- 12738475 TI - Mortality and follow-up colonoscopy after colorectal cancer. AB - OBJECTIVE: There have been no studies that demonstrate surveillance colonoscopy decreases mortality in patients with a history of colorectal cancer. The purpose of this study was to compare the mortality of patients with colorectal cancer who received at least one colonoscopy after their diagnosis with patients who had no further procedures after adjusting for age, race, chemotherapy, radiation therapy, and comorbidity using the national Veterans Affairs (VA) databases. METHODS: We studied a cohort of 3546 patients within the VA national databases with a new diagnosis of colorectal cancer during fiscal year 1995-1996. Patients with inflammatory bowel disease, metastatic disease at presentation, or who died within 1 yr of initial diagnosis were excluded. We collected data for demographics, comorbidities, colonoscopies, chemotherapy, and radiation therapy. The primary outcome was adjusted 5-yr mortality. RESULTS: In the adjusted analysis, the risk of death was decreased by 43% (hazard ratio = 0.57, 95% CI = 0.51-0.64) in the group who had at least one follow-up colonoscopy compared with patients who had no follow-up colonoscopies. CONCLUSIONS: This study strongly supports a mortality benefit for follow-up colonoscopy in patients with a history of nonmetastatic colorectal cancer. PMID- 12738476 TI - Etiology of hepatocellular carcinoma influences clinical and pathologic features but not patient survival. AB - OBJECTIVE: We investigated the relation between hepatocellular carcinoma (HCC) etiology and biological and clinical parameters indicative of severity of liver disease and/or tumor characteristics and patient survival. METHODS: We prospectively recruited 384 patients (82.3% male) with first diagnosis of HCC from 1995 to 1998 in Brescia, Italy. Etiology was assessed by interviewing patients regarding their history of alcohol intake and by testing sera for hepatitis B surface antigen and anti-hepatitis C virus (HCV) antibodies and HCV RNA. RESULTS: Heavy alcohol intake (>60 g of ethanol per day for at least 1 decade) was found in 33.1% of cases, hepatitis B virus (HBV) infection in 9.4%, HCV in 19.8%, hemochromatosis in 1.3%, alcohol and HBV in 12.0%, alcohol and HCV in 16.1%, HBV and HCV in 3.1%, and no factor in 5.2%. Patients with HBV infection with or without heavy alcohol intake were significantly younger than the others (61.7 vs 64.7 yr, p < 0.001). The proportion of males was significantly higher in patients with heavy alcohol intake alone than in the other patient groups (93.7% vs 77.3%, p < 0.001). Among patients with HCV infection with or without heavy alcohol intake, fewer patients had maximum tumor diameter > 5 cm than the others (12% vs 29.1%, p < 0.001). Eighty patients (20.8%) were alive at the end of follow-up (median survival, 17.7 months), and no differences were observed in survival rates by HCC risk factor. CONCLUSIONS: Although some differences were observed in severity of liver disease or tumor characteristics according to etiology, patient survival was not influenced by HCC etiology. PMID- 12738477 TI - Screening for colorectal cancer on the front line. AB - OBJECTIVE: The aim of this study was to assess knowledge, beliefs, and practices of primary care clinicians regarding colorectal cancer screening. METHODS: We surveyed 77 primary care providers in six clinics in central Massachusetts to evaluate several factors related to colorectal cancer screening. RESULTS: Most agreed with guidelines for fecal occult blood test (97%) and sigmoidoscopy (87%), which were reported commonly as usual practice. Although the majority (86%) recommended colonoscopy as a colorectal cancer screening test, it was infrequently reported as usual practice. Also, 36% considered barium enema a colorectal cancer screening option, and it was rarely reported as usual practice. Despite lack of evidence supporting effectiveness, digital rectal examinations and in-office fecal occult blood test were commonly reported as usual practice. However, these were usually reported in combination with a guideline-endorsed testing option. Although only 10% reported that fecal occult blood test/home was frequently refused, 60% reported sigmoidoscopy was. Frequently cited patient barriers to sigmoidoscopy compliance included fear the procedure would hurt and that patients assume symptoms occur if there is a problem. Perceptions of health systems barriers to sigmoidoscopy were less strong. CONCLUSIONS: Most providers recommended guideline-endorsed colorectal cancer screening. However, patient refusal for sigmoidoscopy was common. Results indicate that multiple levels of intervention, including patient and provider education and systems strategies, may help increase prevalence. PMID- 12738478 TI - Orlistat in the treatment of NASH: a case series. AB - Nonalcoholic steatohepatitis is now recognized as a common chronic liver disorder. Up to 16% of affected patients may progress to cirrhosis. The incidence and prevalence of this disease are noted to be increasing, in parallel with the nationwide increase in obesity and diabetes. Treatment options for these patients remain quite limited, however. Weight reduction has been advocated, but there are little data to support this practice, as most patients are unable to comply with the proper dietary modifications. We report three obese patients with biopsy proven nonalcoholic steatohepatitis treated for 6-12 months with a weight reduction medication, orlistat, who lost between 22-42 lb, and had significant clinical and histopathological improvement on follow-up. PMID- 12738483 TI - Re: Dutta et al.-gallbladder cancer. PMID- 12738484 TI - Occult HBV infection in patients with serological markers of past HBV infection. PMID- 12738485 TI - Globus pharyngis: was it a stroke of lightning? PMID- 12738486 TI - Retinal vein thrombosis after infliximab (Remicade) treatment for Crohn's disease. PMID- 12738487 TI - Acute reversible rhabdomyolysis during interferon alpha2B therapy for hepatitis C. PMID- 12738488 TI - Celiac disease in patients with HCV genotype 1A. PMID- 12738489 TI - Necrobiotic nodules: a rare pulmonary manifestion of Crohn's disease. PMID- 12738490 TI - Medication decisions in giant Zenker's and esophageal diverticula. PMID- 12738491 TI - Use of a computerized GI sound analysis system. PMID- 12738492 TI - Relationship between diabetes mellitus and the site of colorectal cancer. PMID- 12738493 TI - Acariasis and disrupted sphincters. PMID- 12738494 TI - Use of recombinant human chorionic gonadotropin in ovulation induction. AB - To review the use of hCG and to describe the clinical benefit of recombinant hCG (r-hCG) based on the published results of prospective, randomized studies. Review of published articles. Tertiary infertility care center.None.None. Oocyte number and quality, luteal phase progesterone, pregnancy and OHSS rate, and local tolerability. The published data consistently show that single doses of 250 microg r-hCG and 5,000 IU urinary (u)-hCG produce similar clinical outcomes when used in infertility treatment cycles for timed intercourse, IUI, and IVF in terms of the number of oocytes retrieved, number of mature oocytes harvested, and fertilization and pregnancy rates attained. Single doses of 10,000 IU u-hCG also gave results comparable to single doses of 250 microg r-hCG. P levels in the midluteal phase were significantly higher with the use of r-hCG compared with u hCG, and local injection site adverse effects were significantly less frequent, demonstrating the higher purity of the recombinant product. A single 500-microg dose of r-hCG led to a higher rate of ovarian hyperstimulation syndrome compared with a 250-microg dose, with no significant improvement in pregnancy rates.A single dose of 250 microg r-hCG was at least as effective as single doses of 5,000 or 10,000 IU u-hCG but offered the advantages associated with use of a recombinant product: local injection site adverse effects were significantly less frequent with r-hCG than with u-hCG. PMID- 12738495 TI - A view from the field on food and drug administration regulation: report of a 2002 survey of U.S. fertility clinics. PMID- 12738496 TI - Cryopreserved embryos in the United States and their availability for research. AB - OBJECTIVE: To determine the number of embryos stored at assisted reproductive technology (ART) clinics in the United States and their current disposition. DESIGN: A targeted survey instrument sent by the SART-RAND team to all medical practices providing in vitro fertilization services in the United States. RESULTS: The SART-RAND team surveyed all 430 ART practices in the United States. Of these practices, 340 returned surveys for analysis. The data from these surveys were merged with data taken from the 1999 SART dataset, which contains information about practice size and success rates. Responding clinics reported a total of 396,526 embryos in storage as of April 11, 2002. The vast majority of the embryos (88.2%) were targeted for patient use. Small numbers of embryos were available for research, donation, destruction, quality assurance, or other uses. CONCLUSIONS: Nearly 400,000 embryos are stored in the United States, the majority of which (88.2%) are targeted for patient use. Few are available for research (2.8%), limiting possible conversion into embryonic stem cell lines. PMID- 12738497 TI - Oral contraceptives improve lung mechanics. AB - OBJECTIVE: To determine whether oral contraceptives affect lung mechanics. DESIGN: Open-label study. SETTING: Academic medical center. PATIENT(S): Thirty six healthy nonsmoking women. INTERVENTION(S): Administration of an oral contraceptive containing ethinyl estradiol, 35 microg, and norgestimat, 250 microg for 6 months. MAIN OUTCOME MEASURE(S): Forced vital capacity; forced expiratory volume in 1 second; peak expiratory flow; and flow at large, medium, and small lung volumes. RESULT(S): At 6 months, all forced expiratory flow and volume had increased significantly (from 6.5% to 15%). Flows at small lung volumes especially increased. CONCLUSION(S): Combination oral contraceptives have a measurable effect on lung mechanics. PMID- 12738498 TI - Transvaginal ultrasonography in women receiving emergency contraception. AB - OBJECTIVE: To determine whether transvaginal ultrasonography improves evaluation of conception time in women seeking emergency contraception. DESIGN: Prospective study. SETTING: Obstetrics and Gynecology Department, Siena University, Siena, Italy. PATIENT(S): One hundred sixty-three women seeking postcoital contraception. MAIN OUTCOME MEASURE(S): Data on menstrual history and time of unprotected intercourse were recorded. Ultrasonographic variables evaluated were ovarian follicle or corpus luteum diameter, endometrial echopattern and thickness, and peritoneal fluid volume. Expected pregnancy rates were calculated according to the probability of conception as estimated from Dixon's table of data, based solely on anamnestic data, or from endometrial or ovarian findings on transvaginal ultrasonography. RESULT(S): According to the menstrual history (cut off level < 0.03) we expected to find 7.6 pregnancies (7.9 in the high-risk group and 0.31 in the low-risk group). According to transvaginal ultrasonography (at the same cut-off), we expected 11.2 pregnancies (0.3 in the low-risk group and 10.9 in the high-risk group). No more than 7 pregnancies were observed, all of which occurred in the high-risk group as determined by transvaginal ultrasonography. In contrast, on the basis of anamnestic data, 4 of 7 pregnancies were in the high-risk group and 3 of 7 were in the low-risk group. CONCLUSION(S): Transvaginal ultrasonography allows timely definition of the fertile period and is a reliable method of computing the day of ovulation. It improves therapeutic options by allowing treatment of only women at high risk of conception. PMID- 12738499 TI - Diagnostic accuracy of laparoscopy, magnetic resonance imaging, and histopathologic examination for the detection of endometriosis. AB - OBJECTIVE: To evaluate the utility of fat-suppressed magnetic resonance imaging (MRI) in the diagnosis of endometriosis. DESIGN: A prospective clinical trial. SETTING: A government research hospital. PATIENT(S): Forty-eight women with pelvic pain. INTERVENTION(S): Magnetic resonance imaging followed by surgical excision and pathologic diagnosis of endometriosis. MAIN OUTCOME MEASURE(S): Presence and extent of endometriosis suggested by preoperative MRIs compared with surgical inspection and biopsy. RESULT(S): A preoperative MRI in 46 women detected fewer endometriosis lesions than histopathology or laparoscopy (78 vs. 101 vs. 150). Few MRI lesions correlated with those identified by laparoscopy (50 of 150) or pathology (38 of 101). Of 42 women with surgically diagnosed endometriosis, 28 had at least one corresponding abnormality on MRI, 5 had abnormalities that didn't correlate with surgical findings, and 9 had normal MRIs. The sensitivity of MRI in detecting biopsy-proven endometriosis for any woman was 69% (25 of 36), and the specificity was 75%. CONCLUSION(S): Although MRI identifies fewer areas of endometriosis than seen at surgery, it suggested endometriosis in 75% of those with at least mild disease. Only 67% of lesions identified at surgery contained histologic evidence of endometriosis. PMID- 12738500 TI - Patient satisfaction and changes in pain scores after ablative laparoscopic surgery for stage III-IV endometriosis and endometriotic cysts. AB - OBJECTIVE: To document the changes in pain scores 3-12 months following ablative laparoscopic surgery. Secondary outcome measures included patient satisfaction scores. DESIGN: A prospective, cohort study. SETTING: A tertiary referral center for the treatment of endometriosis. PATIENT(S): Seventy-three consecutive women with stage III-IV endometriosis and an endometrioma >2 cm. INTERVENTION(S): A laparoscopy was performed. The extraovarian endometriosis was ablated with a CO(2) laser, and the endometrioma capsule was fenestrated then ablated with the potassium-titanic-phosphate (KTP) laser or the Bicap bipolar diathermy. MAIN OUTCOME MEASURE(S): Pre- and postoperative visual analogue scores for pelvic pain were completed. Patient satisfaction was scored from 1 to 10, with a score of 10 being "most satisfied." RESULT(S): A total of 73 women with stage III-IV endometriosis and 96 cysts (23 cysts were bilateral). The mean revised American Fertility Society (AFS) score was 65.5 (range 22-128). At 12 months, the mean temporal decrease in the pain score for dyspareunia was 2.14 +/- 0.41; for dysmenorrhea, 1.52 +/- 0.38; and for chronic nonmenstrual pain, 2.37 +/- 0.43. Sixty-four (87.7%) patients were satisfied or very satisfied with the treatment. No surgical complications occurred. CONCLUSION(S): Laparoscopic ablative surgery for endometriomas in the presence of stage III-IV endometriosis is an effective treatment for relieving pelvic pain. PMID- 12738501 TI - Performance of basal follicle-stimulating hormone in the prediction of poor ovarian response and failure to become pregnant after in vitro fertilization: a meta-analysis. AB - OBJECTIVE: To assess the predictive performance and clinical value of basal FSH as a test for ovarian reserve in in vitro fertilization (IVF) patients. DESIGN: Meta-analysis. SETTING: Tertiary fertility center. PATIENT(S): Patients undergoing IVF. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Poor ovarian response, nonpregnancy. RESULT(S): We located 21 studies that had reported on basal FSH and IVF outcome. No single study met high standards of methodological rigor; most studies are of moderate methodological quality only. The summary receiver operating characteristic curve indicated a moderate predictive performance for poor response, and a low predictive performance for nonpregnancy. Predictions with a substantial shift from pre-FSH-test probability to post-FSH test probability are only achieved at extreme cut-off levels for basal FSH. Sensitivity of such cut-off levels, for both the prediction of poor response and nonpregnancy, is limited. CONCLUSION(S): Clinical value of testing for basal FSH is restricted to a small minority of patients. Basal FSH should not be regarded as a useful routine test for the prediction of IVF outcome. The development of better tests to assess ovarian reserve remains of importance. PMID- 12738503 TI - Effect of blastomere loss on the outcome of frozen embryo replacement cycles. AB - OBJECTIVE: To assess the impact of survival of cryopreservation and thawing with all blastomeres intact on the outcome of frozen embryo replacement (FER) cycles. DESIGN: Prospective observational study. SETTING: University-affiliated tertiary referral assisted conception unit. PATIENT(S): The number of intact blastomeres before cryopreservation and after thawing was prospectively recorded in 1,687 cleavage-stage embryos thawed in 377 FER cycles. The cycles were categorized into two groups: group A (n = 184) included cycles in which all embryos transferred survived the cryopreservation and thawing process with all their original blastomeres intact; group B (n = 193) included cycles in which embryos transferred included at least one partially damaged embryo that has lost up to 50% of its original blastomere number. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Pregnancy and embryo implantation rates. RESULT(S): Groups A and B were comparable with respect to mean age at cryopreservation, mean number of oocytes retrieved and fertilized normally in the fresh cycle, and mean age at frozen transfer. No significant difference was found between the two groups with regard to mean number of frozen and thawed embryos per cycle and mean endometrial thickness reached before P supplementation. More embryos were transferred per cycle in group B than group A (2.4 +/- 0.6 vs. 2.1 +/- 0.6, respectively). However, the pregnancy and clinical pregnancy rates per cycle were significantly higher in group A than in group B (39.1% and 28.3% vs. 22.8% and 13.5%, respectively). The implantation rate was also higher in group A than in group B (17.3% vs. 8.1%, respectively). CONCLUSION(S): FER cycles in which all embryos transferred remained fully intact at thawing achieve a better outcome than those with at least one partially damaged embryo. PMID- 12738502 TI - Comparison of changes in uterine contraction frequency after ovulation in the menstrual cycle and in in vitro fertilization cycles. AB - OBJECTIVE: To compare changes in uterine contraction (UC) frequency occurring after ovulation in the menstrual cycle and in IVF and to clarify UC anomalies encountered in IVF. DESIGN: Prospective crossover trial. SETTING: University infertility clinic. PATIENT(S): Six women seeking IVF for male factor infertility having regular menstrual cycles were studied during the menstrual cycle and IVF. INTERVENTION(S): Identification of LH surge in the menstrual cycle. Measurement of plasma E(2) and P levels and assessment of UC frequency by direct ultrasound visualization on the day of LH surge in the menstrual cycle and the day of hCG administration in IVF and every 2 days thereafter for 6 days. MAIN OUTCOME MEASURE(S): Uterine contraction (UC) frequency and plasma E(2) and P levels. RESULT(S): UC frequency was similar on the day of LH surge (5/minute) and hCG administration (5.3/minute). Establishment of uteroquiescence was more prompt in the menstrual cycle than in IVF, with lower UC frequency 4 days after LH surge compared with 4 days after hCG, while E(2) and P levels were higher in IVF. Six days after LH surge/hCG administration, UC frequency was low in the menstrual cycle and in IVF. CONCLUSION(S): High UC frequency in IVF at the time of ET results from delayed establishment of uteroquiescence after ovulation in IVF as compared with the menstrual cycle. In IVF, low UC frequency 6 days after hCG may contribute to the higher pregnancy rates observed with blastocyst transfers. PMID- 12738504 TI - Effect of soy-derived isoflavones on hot flushes, endometrial thickness, and the pulsatility index of the uterine and cerebral arteries. AB - OBJECTIVE: To determine the effect of soy-derived isoflavones on hot flushes, endometrial thickness, and the vascular reactivity of uterine and cerebral arteries. DESIGN: Double-blind, randomized, placebo-controlled trial. SETTING: Healthy volunteers in an academic research environment. PATIENT(S): Sixty-two postmenopausal women aged 45-60 years attending the Outpatient Menopause Clinic of our gynecological departments. INTERVENTION(S): The patients were administered 72 mg of soy-derived isoflavones or placebo under double-blind conditions. The daily number of hot flushes was recorded in a diary. Endometrial thickness was measured by means of transvaginal ultrasound; the uterine, internal carotid, and middle cerebral arteries were evaluated using Doppler ultrasound. MAIN OUTCOME MEASURE(S): The daily number of hot flushes, endometrial thickness, and arterial pulsatility index (PI). RESULT(S): Both treatments led to a 40% reduction in the number of hot flushes. Soy-derived isoflavones had no effect on endometrial thickness or the PI of the uterine and cerebral arteries. CONCLUSION(S): The daily administration of 72 mg of soy-derived isoflavones is no more effective than placebo in reducing hot flushes in postmenopausal women. It also has no effect on endometrial thickness or the PI of the uterine and cerebral arteries. PMID- 12738505 TI - Effect of short-term hormone therapy on oxidative stress and endothelial function in African American and Caucasian postmenopausal women. AB - In postmenopausal women (PMW), the effect of a short-term course of estrogen/progestin HT on free radical oxidative stress was evaluated. In addition, HT's effect on plasma nitric oxide (NO) activity was determined as a measure of vascular endothelial function. We investigated the relationship of these markers and HT across race and the cardiovascular risk factors of smoking, diabetes and hypertension.A prospective, observational study comparing preintervention and postintervention. Academic research center.Twenty-seven (14 African American and 13 Caucasian) PMW volunteers. Six weeks of continuous, combined estrogen/progestin HT. Plasma concentrations of free 8-epi-prostaglandin F(2alpha) (8-isoprostane) before and after HT were compared as a measure of oxidative stress. Nitrite, the stable oxidation metabolite of NO, was measured by the Greiss reaction after nitrate reduction to nitrite with cadmium. Plasma levels of free 8-isoprostane decreased significantly after 6 weeks of HT. Although almost all subjects benefited from the reduction in free 8-isoprostane, PMW with at least one cardiovascular risk factor (n = 19) demonstrated higher free 8-isoprostane than did subjects with no risk factors. Plasma levels of nitrite increased after 6 weeks of HT, but the difference was not statistically significant. Caucasian PMW demonstrated a greater increase in plasma levels of nitrite after 6 weeks of HT as compared with African American subjects, who exhibited almost no change.Short-term administration of HT significantly reduces oxidative stress in PMW and is consistent across race. However, there was an observed racial difference in endothelial NO response to HT between African American and Caucasian PMW. PMID- 12738506 TI - Lower levels of inhibin A and pro-alphaC during the luteal phase after triggering oocyte maturation with a gonadotropin-releasing hormone agonist versus human chorionic gonadotropin. AB - OBJECTIVE: To investigate the effect of triggering oocyte maturation with GnRH agonist on corpus luteum function by measuring luteal phase levels of inhibin A and pro-alphaC. DESIGN: Prospective randomized trial. SETTING: In vitro fertilization (IVF) program at a university hospital. PATIENT(S): Infertile women undergoing IVF-ET treatment. INTERVENTION(S): Controlled ovarian hyperstimulation with FSH and GnRH antagonist, triggering of final oocyte maturation with either hCG (n = 8) or GnRH agonist (n = 8), IVF-ET, and collection of blood samples every 2-3 days during the luteal phase. MEASUREMENTS AND MAIN RESULTS: Luteal phase serum levels of inhibin A and pro-alphaC, P, and E(2). RESULT(S): Levels of inhibin A, pro-alphaC, estrogen, and P were significantly lower from day 4 to day 14 after triggering final oocyte maturation by GnRH agonist compared with hCG. Maximal luteal serum inhibin A and pro-alphaC levels were 91.5 +/- 23.6 and 184.1 +/- 23.5 pg/mL in the GnRH agonist-treated women compared with 464.7 +/- 209.1 and 7,351.6 +/- 934.3 pg/mL in women treated with hCG. CONCLUSION(S): Triggering final oocyte maturation with GnRH agonist instead of hCG in IVF cycles dramatically decreases luteal levels of inhibins, reflecting significant inhibition of the corpus luteum function. This effect may explain, at least in part, the mechanism of ovarian hyperstimulation syndrome prevention by the use of GnRH agonist. PMID- 12738507 TI - Effect of pituitary down-regulation on the ovary before in vitro fertilization as measured using three-dimensional power Doppler ultrasound. AB - OBJECTIVE: To evaluate the changes taking place in the ovaries during pituitary down-regulation. DESIGN: Prospective observational study of women undergoing IVF treatment. SETTING: A tertiary referral center for assisted reproduction. PATIENT(S): Forty women who received the long buserelin acetate treatment protocol. Transvaginal three-dimensional power Doppler ultrasound examinations before and after pituitary down-regulation. INTERVENTION(S): Ovarian volume, number of follicles, vascularization index (VI), flow index (FI), vascularization flow index (VFI), and mean gray value (MG). RESULT(S): Before the pituitary down regulation, the dominant ovary was larger in volume and had a lower MG than the nondominant ovary. After the down-regulation, there was a significant decrease in the volume and number of follicles and an increase in MG. After pituitary down regulation, the dominant and nondominant ovaries did not differ from each other in any of the parameters. Polycystic ovaries were larger than normal ones before and after the down-regulation, without any differences in MG, VI, FI, or VFI. Right and left ovaries did not differ from each other after the down-regulation. CONCLUSION(S): The differences observed between dominant and nondominant ovaries seem to disappear after pituitary down-regulation. In addition, polycystic ovaries were always larger than the normal ones, but no differences could be detected in the stromal brightness or vascularity either before or after the administration of GnRH agonist therapy. PMID- 12738508 TI - Obesity and the risk of spontaneous abortion after oocyte donation. AB - OBJECTIVE: To determine whether obesity increases the risk of spontaneous abortion. DESIGN: Retrospective study. SETTING: Oocyte donation program at the Instituto Valenciano de Infertilidad in Spain. PATIENT(S): Seven hundred twelve cycles of recipients of ovum donation with known body mass index (BMI), good quality embryo transfer, and absence of uterine pathology or clinical history of antiphospholipid antibodies or recurrent abortion. INTERVENTION(S): Recipients were divided in four BMI (kg/m(2)) groups: lean, with BMI <20 (n = 92; 12.9%); normal, with BMI = 20-24.9 (n = 398; 55.9%); overweight, with BMI = 25-29.9 (n = 172; 24.2%); and obese, with BMI >/=30 (n = 50; 7%). Clinical parameters were compared among the groups. MAIN OUTCOME MEASURE(S): Spontaneous abortion rates according to BMI. RESULT(S): No difference was found among the four BMI groups in any of the parameters of the cycle analyzed. The overall abortion rate was 15.8% (57 of 360). There were significant differences in abortion rates between the obese (38.1%), and the normal (13.3%) and overweight (15.5%) groups. When several cutoff BMI values were established (20, 25, and 30), only the obese women demonstrated a greater risk of abortion. Compared with the normal population, the obese group showed a significant fourfold increase in the risk of spontaneous abortion. CONCLUSION(S): Our findings confirm that obesity (BMI >/=30) is an independent risk factor for spontaneous abortion. Therefore, it would be advisable for obese patients to reduce weight before becoming pregnant. PMID- 12738509 TI - Combined thrombophilic polymorphisms in women with idiopathic recurrent miscarriage. AB - OBJECTIVE: To identify associations or interrelations between carriage of the methylenetetrahydrofolate reductase (MTHFR) C677T, the MTHFR A1298C, the factor V Leiden G1691A, the factor II prothrombin G20210A, the human platelet antigen (HPA) 1 C12548T, and the apolipoprotein (APO) B R3500Q polymorphisms and idiopathic recurrent miscarriage (IRM). DESIGN: Prospective case control study. SETTING: Academic research institution. PATIENT(S): One hundred forty-five women with a history of three or more consecutive pregnancy losses before 20 weeks gestation and 101 healthy postmenopausal women with at least two live births and no history of pregnancy loss. INTERVENTION(S): Peripheral venous punctures. MAIN OUTCOME MEASURE(S): Multiplex polymerase chain reaction was performed to identify the different alleles of six candidate genetic risk factors for IRM (MTHFR C677T, MTHFR A1298C, factor V Leiden G1691A, factor II prothrombin G20210A, HPA 1 C12548T, and the APO B R3500Q). RESULT(S): Allele and genotype frequencies of all polymorphisms were not significantly different between the study and the control groups. Also, no significant associations occurred between combinations of polymorphisms and the occurrence of IRM. CONCLUSION(S): Our data fall short of showing any significant association between single polymorphisms of the MTHFR, the Factor V Leiden, the Factor II Prothrombin, the HPA 1 and APO B genes or combinations of these polymorphisms and the occurrence of IRM. PMID- 12738511 TI - Aging women with polycystic ovary syndrome who achieve regular menstrual cycles have a smaller follicle cohort than those who continue to have irregular cycles. AB - OBJECTIVE: To examine whether follicle loss due to ovarian aging is responsible for the occurrence of regular menstrual cycles in aging women with polycystic ovary syndrome (PCOS), the size of the FSH-sensitive follicle cohort was estimated by the exogenous follicle-stimulating hormone ovarian reserve test (EFORT) and related to the follicle count as measured by ultrasound. DESIGN: Prospective study. SETTING: Reproductive endocrinology unit of an academic medical center. PATIENT(S): Twenty-seven aging women with PCOS (35.8-49.4 years): 20 with regular menstrual cycles and 7 with oligomenorrhea or amenorrhea. INTERVENTION(S): EFORT and transvaginal ultrasound. MAIN OUTCOME MEASURE(S): Baseline (cycle day 2, 3, or 4) FSH, androstenedione (A), T, E(2), and inhibin B levels, the E(2) and inhibin B increment after the EFORT, and the follicle count. RESULT(S): After correction for the body mass index (BMI), the inhibin B increment was higher in the irregular menstrual group, but the E(2) increment did not differ significantly between the two groups. Ultrasound showed a median follicle count of 8.5 (4.0-18.0) in women with regular menstrual cycles (n = 16), compared with 18.0 (8.0-35.0) in irregularly menstruating women (n = 7). The follicle count was significantly correlated to the FSH-induced E(2) increment (r = 0.656) as well as to the inhibin B increment (r = 0.654). The regularly menstruating group was significantly older, had a higher basal FSH concentration, and had lower androgens than the irregularly menstruating group. CONCLUSION(S): The smaller follicle count, the older age, the higher FSH concentration, and the lower FSH-induced inhibin B increment found in women with PCOS and a regular menstrual cycle confirm that a decrease in the size of the follicle cohort due to ovarian aging is largely responsible for the regular menstrual cycles in aging PCOS women. PMID- 12738510 TI - Localization and cellular distribution of pregnancy-associated plasma protein-a and major basic protein in human ovary and corpora lutea throughout the menstrual cycle. AB - OBJECTIVE: To assess the expression and cellular distribution of pregnancy associated plasma protein-A (PAPP-A) and major basic protein (MBP) in human ovarian tissue during the menstrual cycle. DESIGN: Ovarian tissues (n = 50) and corpora lutea (n = 18) were obtained from patients undergoing hysterectomy/oophorectomy for benign conditions and tissue sections were immunostained for MBP and PAPP-A. SETTING: University medical center. INTERVENTION(S): Immunostaining of tissue sections using antibodies to PAPP-A and MBP. MAIN OUTCOME MEASURE(S): Microscopic evaluation to assess the presence, distribution, and cellular co-localization of MBP and PAPP-A and to describe any variations in their expression during the menstrual cycle. RESULT(S): Major basic protein (MBP) is found in several ovarian cell types throughout the menstrual cycle. The MBP immunostaining of ovarian follicles varied depending on the size, with primordial follicles staining in the ooplasm with a lack of staining in the granulosa and theca cells. In the intermediate/mature follicles, MBP was immunolocalized in theca, but not in granulosa cells except in the mature follicles. Pregnancy-associated plasma protein-A (PAPP-A) was immunolocalized in primordial follicle ooplasm, theca externa of intermediate/mature follicles, and in granulosa cells with increased intensity as luteinization progressed. The luteal tissue is the major site of MBP and PAPP-A with highest intensity found during the midluteal phase associated with both small and large luteal cells. CONCLUSION(S): The expression and distinct pattern of MBP and PAPP-A cellular localization in human ovarian tissue during folliculogenesis and in luteal tissue suggest that their individual and combined actions in a cell specific fashion may play a role in growth and differentiation of theca, granulosa, and luteal cells. PMID- 12738512 TI - Regulation of transforming growth factor-beta, type III collagen, and fibronectin by dichloroacetic acid in human fibroblasts from normal peritoneum and adhesions. AB - OBJECTIVE: To examine the role of aerobic metabolism in fibroblasts from normal peritoneum and adhesions in the differential expression of extracellular matrix (ECM) and transforming growth factor-beta (TGF-beta), an inflammatory cytokine that regulates ECM expression. DESIGN: Cell culture under normoxic and hypoxic conditions. SETTING: University research laboratory. PATIENT(S): Human fibroblasts cultures from normal peritoneum and adhesions. INTERVENTION(S): Exposure to dichloroacetic acid (DCA), which activates pyruvate dehydrogenase, for 24 hours under normal and hypoxic (2% O(2)) conditions. MAIN OUTCOME MEASURE(S): Multiplex reverse transcriptase polymerase chain reaction (RT/PCR) of type III collagen, fibronectin, TGF-beta1, and beta-actin was performed, with analysis of PCR-amplified products performed by densimetric analysis of gel bands using the National Institutes of Health Image analysis program. RESULT(S): DCA inhibited human peritoneal fibroblast and adhesion fibroblast TGF-beta1 mRNA expression under normoxic and hypoxic conditions. DCA also markedly reduced fibronectin and type III collagen expression under hypoxic conditions in fibroblasts from normal peritoneum and adhesions. CONCLUSION(S): These observations provide further support for the suggestion that regulation of metabolic activity of peritoneal cells may provide a target for interventions designed to reduce postoperative adhesions. PMID- 12738513 TI - Peritoneal fluid concentrations of matrix metalloproteinase-9, tissue inhibitor of metalloproteinase-1, and transforming growth factor-beta in women with pelvic adhesions. AB - OBJECTIVE: To establish whether the concentration of matrix metalloproteinase-9 (MMP-9), tissue inhibitor of metalloproteinase-1 (TIMP-1), and transforming growth factor-beta (TGF-beta) is influenced by the presence or absence of adhesions, and whether the concentration of these mediators vary throughout the menstrual cycle. DESIGN: Prospective case-control study. SETTING: Women undergoing laparoscopy in a university hospital in the United Kingdom. PATIENT(S): Women undergoing laparoscopy for benign gynecological conditions. INTERVENTION(S): Samples of peritoneal fluid were collected at diagnostic laparoscopy in one group, and at laparoscopy and serially during the first 48 hours after laparoscopic adhesiolysis in a second group. We correlated the concentrations of mediators in serially sampled peritoneal fluid during the 48 hours following laparoscopic adhesiolysis to the adhesion formation and reformation found during second-look laparoscopy. MAIN OUTCOME MEASURE(S): The concentrations of MMP-9, TIMP-1, and TGF-beta in peritoneal fluid. RESULT(S): MMP 9 concentration was lower in the follicular phase than the luteal phase of the menstrual cycle. MMP-9 concentration was significantly lower in women with pelvic adhesions than in women with a normal pelvis. The MMP-9/TIMP-1 ratio is significantly higher in women with significant adhesions at second-look laparoscopy compared to women with minimal or no adhesions. CONCLUSION(S): The components of extracellular matrix remodeling may play an important part in the adhesion formation/reformation process. PMID- 12738514 TI - Cytokine microenvironments in human first trimester decidua are dependent on trophoblast cells. AB - OBJECTIVE: To compare cytokine expression profiles of decidua basalis (containing trophoblast cells) and decidua parietalis (without trophoblast cells) for determination of microenvironments in human first trimester decidua. DESIGN: Retrospective study. SETTING: School of Medicine, RWTH University of Aachen, Aachen Germany, and Bourgognekliniek Maastricht, Maastricht, The Netherlands. PATIENT(S): Forty-six women who had undergone elective first-trimester termination of viable pregnancy at 5 to 12 weeks. MAIN OUTCOME MEASURE(S): Quantitative cytokine protein analysis in decidual tissues by enzyme-linked immunosorbent assay, qualitative cytokine messenger (m)RNA analysis in isolated decidual cell samples, and comparative mRNA and protein analysis in tissues of decidua basalis compared with decidua parietalis. RESULT(S): Interleukin-2, interferon-gamma (Th-1), interleukin-4 (Th-2), and interleukin-1beta proteins are expressed in the human first-trimester decidua. Interleukin-2, interferon-gamma, and interleukin-4 mRNA mainly derive from the decidual tissue leukocytes. Interleukin-1beta mRNA is expressed by all decidual cell types. Interferon-gamma mRNA and protein is detected predominantly in the decidua basalis, which contains trophoblast cells. CONCLUSION(S): Microenvironments are established topographically by different expression of cytokines in decidua basalis and decidua parietalis. These locally specific patterns are indicative of fetomaternal cross-talk. Higher interferon-gamma concentrations in decidua basalis may influence leukocyte differentiation (e.g., macrophage activation) and trophoblast invasion (e.g., by induction of expression of major histocompatibility complex). PMID- 12738515 TI - Messenger RNA expression for the follicle-stimulating hormone receptor and luteinizing hormone receptor in human oocytes and preimplantation-stage embryos. AB - OBJECTIVE: To study the expression of the FSH and LH receptors in human oocytes and preimplantation embryos and their potential roles in early human development. DESIGN: Clinical and molecular studies. SETTING: University hospital IVF center. PATIENT(S): Female volunteers undergoing intracytoplasmic sperm injection at the IVF unit of the Athens University Hospital. All patients gave written informed consent. INTERVENTION(S): Ovarian stimulation was performed with exogenous gonadotropin administration. Intracytoplasmic sperm injection was performed on mature oocytes. MAIN OUTCOME MEASURE(S): Unfertilized oocytes and zygotes and embryos at the 2-cell, 4-cell, morula, and blastocyst stage were selected for study. A polymerase chain reaction methodology was used to analyze human oocytes and embryos. Messenger RNA was reverse transcribed and amplified with FSH and LH receptor specific primers. RESULT(S): Transcripts for the FSH receptor were detected in oocytes and zygotes and embryos at the 2-cell, morula, and blastocyst stage, while no message was detected in embryos at the 4-cell stage. Transcripts for the LH receptor were observed in oocytes and zygotes and morula- and blastocyst-stage embryos, whereas no message was detected in embryos at the 2 cell and 4-cell stage. CONCLUSION(S): Messenger RNA for the FSH and LH receptors was observed in oocytes and preimplantation embryos at different stages, indicating a physiological role in the oocyte maturation process and early embryonic development in the human. PMID- 12738516 TI - Use of a levonorgestrel-releasing intrauterine system to treat bleeding related to uterine leiomyomas. AB - OBJECTIVE: This study was designed to evaluate the potential usefulness of the levonorgestrel-releasing intrauterine system (LNG IUS) in treating women with uterine leiomyomas. DESIGN: Prospective before-and-after study. SETTING: Family planning unit in an academic research institute. PATIENT(S): Sixty-seven women with uterine leiomyomas who chose the LNG IUS as their method of contraception. INTERVENTION(S): Clinical and ultrasound examinations were performed prior to and 3, 6, and 12 months after the LNG IUS insertion. MAIN OUTCOME MEASURE(S): Menstrual blood loss assessed with pictorial blood loss assessment charts, ferritin and hemoglobin concentrations, and uterine and leiomyoma volume. RESULT(S): Use of the LNG IUS was associated with a marked reduction in menstrual blood loss. After 12 months of use, the mean pictorial blood loss assessment chart score declined from 97 to 16 (P<.001). Hemoglobin and ferritin levels increased significantly over 1 year of use. Eighteen of 19 women (95%) who were anemic at the beginning of the study were no longer anemic at 12 months, as judged by hemoglobin levels. No pregnancies occurred during the study. CONCLUSION(S): The LNG IUS was associated with a profound reduction in menstrual blood loss. For women with leiomyomas of this size, the LNG IUS provides effective medical treatment of bleeding. PMID- 12738517 TI - Microsurgical resection of nonocclusive salpingitis isthmica nodosa is beneficial. AB - OBJECTIVE: To determine the effect of microsurgical resection and tubocornual anastomosis (TCA) of nonocclusive salpingitis isthmica nodosa (SIN) on fertility and risk for ectopic pregnancy (EP). DESIGN: Prospective cohort. SETTING: University-affiliated tertiary fertility clinic. PATIENT(S): Infertile women with hysterosalpingography evidence of SIN in patent fallopian tubes. INTERVENTION(S): Microsurgical resection and TCA for nonocclusive SIN. MAIN OUTCOME MEASURE(S): Occurrence of IUP and EP after TCA; comparison of duration of infertility preceding TCA with time to intrauterine pregnancy (IUP) after TCA; and comparison of numbers of women who conceived an EP before and after TCA. RESULT(S): Twelve (46%) of the women had IUPs with a mean time to pregnancy of 10.5 months, which is significantly shorter than the preceding period of infertility. Three women experienced EPs after TCA, which is reduced compared with the number of women with an EP preceding the TCA. CONCLUSION(S): The significant decrease in time to conceive an IUP after surgery as compared with the duration of infertility before surgery and the apparent reduction in risk for EP after surgery demonstrate the benefit of TCA for resection of nonocclusive SIN. PMID- 12738519 TI - Use of a single preoperative dose of misoprostol is efficacious for patients who undergo abdominal myomectomy. AB - OBJECTIVE: To investigate the effectiveness of a single preoperative dose of misoprostol in abdominal myomectomies. DESIGN: Placebo-controlled randomized prospective study. SETTING: Department of obstetrics and gynecology in a university hospital. PATIENT(S): Twenty-five women with symptomatic uterine leiomyomas. INTERVENTION(S): Among patients undergoing abdominal myomectomies, an hour before the operation women in the study group (n = 13) were given a single dose of vaginal misoprostol (400 microg); those in the control group (n = 12) were given placebo. MAIN OUTCOME MEASURE(S): Intraoperative blood loss, duration of operation, duration of postoperative hospitalization, and the need for blood transfusion were compared between the control and study groups. RESULT(S): Blood loss, operation time, and need for postoperative blood transfusion were significantly reduced in the group given vaginal misoprostol. No difference was observed among patients in terms of the time of hospitalization. CONCLUSION(S): A single preoperative dose of vaginal misoprostol is a simple, reliable method for reducing intraoperative blood loss and need for postoperative blood transfusion after abdominal myomectomies. PMID- 12738518 TI - A new technique of laparoscopic ovariopexy before irradiation. AB - OBJECTIVE: To report a new technique of laparoscopic ovarian transposition to preserve ovarian function in women who require pelvic irradiation for musculoaponeurotic fibromatosis (extra abdominal desmoid). DESIGN: Case report. SETTING: University teaching hospital. PATIENT(S): Two nulliparous women who required adjunctive radiotherapy for musculoaponeurotic fibromatosis where radiotherapy planning indicated that the right ovary could be removed from the field of radiation by anterior transposition. INTERVENTION(S): Laparoscopic suturing of the right ovary to the right round ligament with intracorporeal polypropylene sutures. MAIN OUTCOME MEASURE(S): Technical feasibility, recovery, postoperative adhesions, ease of ovarian repositioning, and evidence of ovulation after completion of radiotherapy. RESULT(S): The technique was easily performed without needing to divide the ligament of the ovary. Recovery was rapid, and there were no postoperative adhesions. The ovary showed evidence of continued function and was easily repositioned by dividing the sutures. CONCLUSION(S): In selected cases, this method of ovarian transposition has the advantages not only of being technically easy but also of allowing for repositioning of the ovary with minimal disruption of its anatomical relationship to the fallopian tube, thereby favoring fertility. PMID- 12738520 TI - Repeated laparoscopic ovarian diathermy is effective in women with anovulatory infertility due to polycystic ovary syndrome. AB - OBJECTIVE: To assess the effectiveness of a second laparoscopic ovarian diathermy (LOD) in women with polycystic ovary syndrome (PCOS). DESIGN: Retrospective study. SETTING: Tertiary infertility clinic. PATIENT(S): Twenty anovulatory infertile women with PCOS who underwent LOD 1-6 years previously. Twelve subjects had previously responded positively to the first LOD, but the anovulatory status recurred (Group I), whereas eight subjects had not responded at all (Group II). INTERVENTION(S): Laparoscopic ovarian diathermy. MAIN OUTCOME MEASURE(S): Rates of ovulation, pregnancy and resumption of menstrual regularity, and biochemical changes. RESULT(S): An overall ovulation rate of 12 out of 20 women (60%) and a pregnancy rate of 10 out of 19 (53%) were achieved after the second LOD. In women (n = 12) who previously responded positively to the first LOD (LOD-sensitive), the ovulation and pregnancy rates were 10 out of 12 (83%) and 8 out of 12 women (67%), respectively, which were significantly (P<.05) higher than 2 out of 8 (25%) and 2 out of 7 (29%) of the previous nonresponders (LOD-resistant). Statistically significant hormonal changes including reduction of luteinizing hormone (LH), testosterone (T), and free androgen index (FAI) after the repeat LOD were only observed in the LOD-sensitive group. CONCLUSION(S): Repeat LOD is highly effective in women who previously responded to the first procedure. PMID- 12738521 TI - Safety of conservative management and fertility outcome in women with borderline tumors of the ovary. AB - OBJECTIVE: To assess the safety of fertility-sparing treatment and the remaining chance of childbearing after surgery. DESIGN: Retrospective clinical study. DESIGN: Gynecology department of a university teaching hospital. PATIENT(S): Seventy-five women underwent surgical management in our institution between 1986 and 2001 for borderline tumors of the ovary. INTERVENTION(S): Fifty-nine patients were treated by radical, fertility-compromising surgery. The remaining 16 patients underwent conservative surgery, preserving the uterus and at least some functional ovarian tissue. Seven unilateral adnexectomies, one simple cystectomy, and two adnexectomies associated with contralateral cystectomy were performed. MAIN OUTCOME MEASURE(S): Recurrence, survival, and pregnancy rates. RESULT(S): The observed recurrence rates after radical and conservative surgery were 0.0% and 18.7%, respectively. No disease-related deaths occurred in any group; there is no significant difference in survival rates. We can report 12 pregnancies in 7 of 11 women who underwent fertility-sparing management and who wished to become pregnant. CONCLUSION(S): In certain circumstances, conservative management offers a safe solution for borderline tumors of the ovary. Recurrence is noted significantly more often after this type of treatment, but all cases of recurrent disease can be detected with close follow-up and can be treated accordingly. No significant change in survival rates was found. Moreover, the pregnancy rate in women desiring pregnancy, those treated conservatively, was as high as 63.6%. PMID- 12738522 TI - A prospective study to evaluate the efficacy of two- and three-dimensional sonohysterography in women with intrauterine lesions. AB - OBJECTIVE: To measure the accuracy of three-dimensional (3D) sonohysterography in detecting intrauterine lesions. DESIGN: Prospective study. SETTING: Teaching hospital. PATIENT(S): Two hundred nine infertile patients suspected to have an intrauterine lesion on 2D ultrasound or hysterosalpingography. INTERVENTION(S): Three-dimensional ultrasound, 2D and 3D sonohysterography (SHG). Ninety-two of the patients had a lesion distorting the endometrium on the 3D SHG, and those were referred for hysteroscopy. MAIN OUTCOME MEASURE(S): Sensitivity and specificity of 2D, 3D ultrasound, and 2D SHG compared to 3D SHG. RESULT(S): Of the 92 patients with a lesion, 48 had polyps, 35 submucous/intramural myomas, 3 both polyps and myomas, 4 mullerian anomalies, 1 thick endometrium, and 1 synechiae. Compared with the 3D SHG results, the sensitivity and specificity were 97% and 11% for the 2D transvaginal ultrasound, 87% and 45% for the 3D ultrasound, and 98% and 100% for the 2D SHG. In the group of 59 patients who had hysteroscopy, the sensitivity of the 2D SHG and 3D SHG were 98% and 100%, with a positive predictive value of 95% and 92%, respectively. CONCLUSION(S): Three dimensional sonohysterography allows precise recognition and localization of lesions. If 2D and 3D SHG are normal, invasive diagnostic procedures such as hysteroscopy could be avoided. PMID- 12738523 TI - Hysteroscopic appearance of the endometrial cavity following thermal balloon endometrial ablation. AB - OBJECTIVE: To assess the appearance of the endometrial cavity after thermal balloon endometrial ablation. DESIGN: Observational study. SETTING: University teaching hospital. PATIENT(S): Twenty-two women who had undergone thermal balloon endometrial ablation and who were followed up for at least 6 months. INTERVENTION(S): Outpatient diagnostic hysteroscopy. MAIN OUTCOME MEASURE(S): Appearance of the endometrial cavity and presence of intrauterine adhesions on hysteroscopy. RESULT(S): Postablation intrauterine adhesions were found in eight women (36.4%); six had focal adhesions in the fundal area and two had complete obliteration of the cavity. Of these eight women, three had spotting during menstruation, three had hypomenorrhea, one had eumenorrhea, and one had amenorrhea. The uterine cavity was fibrotic in four (18%) women; all reported spotting during menstruation. Ten women had a normal uterine cavity; eight had hypomenorrhea, one had spotting, and one had eumenorrhea. CONCLUSION(S): The hysteroscopic appearance of the uterine cavity after thermal balloon endometrial ablation varies considerably. Menstrual outcome is associated with postablation appearance. PMID- 12738524 TI - Surrogate pregnancy in a patient who underwent radical hysterectomy and bilateral transposition of ovaries. AB - OBJECTIVE: To evaluate IVF-surrogate pregnancy in a patient with ovarian transposition after radical hysterectomy for carcinoma of the cervix. DESIGN: Case report. SETTING: A maternity hospital in Tel Aviv that is a major tertiary care and referral center. PATIENT(S): A 29-year-old woman who underwent Wertheim's hysterectomy for carcinoma of the uterine cervix and ovarian transposition before total pelvic irradiation. INTERVENTION(S): Standard IVF treatment, transabdominal oocyte retrieval, and transfer to surrogate mother. MAIN OUTCOME MEASURE(S): Outcome of IVF cycle. RESULT(S): A twin pregnancy in the first cycle. CONCLUSION(S): This is the second reported case of controlled ovarian stimulation and oocyte retrieval performed on a transposed ovary. PMID- 12738525 TI - Successful triplet pregnancy and delivery after oocyte donation in an infertile female with chromosome mosaicism for monosomy X, partial trisomy X, and terminal Xp deletion. AB - OBJECTIVE: To present successful triplet pregnancy and delivery after oocyte donation and IVF in a female with chromosome mosaicism for monosomy X, partial trisomy X, and terminal Xp deletion. DESIGN: Descriptive case study. SETTING: Large tertiary care hospital. PATIENT(S): A 23-year-old infertile woman with primary amenorrhea, short stature, and a mosaic karyotype of 45,X[10]/46,X,idic(X)(qter-->p22.3::p22.3-->qter)[40]. Fluorescence in situ hybridization investigation of the isodicentric X chromosome revealed terminal Xp deletion. INTERVENTION(S): Ultrasound and laparoscopic examinations revealed streak ovaries and a hypoplastic uterus. Oocyte donation and IVF resulted in a triplet pregnancy without major maternal complications. MAIN OUTCOME MEASURE(S): Ultrasound and laparoscopy. RESULT(S): A cesarean section was performed at 32 weeks gestation because of preterm labor and malpresentation. Three babies were delivered with birth weights of 1,514 g, 1,686 g, and 1,968 g. All infants survived and were healthy at 3 years of age. CONCLUSION(S): With careful evaluation and counseling, assisted reproductive technology can be safely used in females with mosaic X chromosome abnormalities and gonadal dysgenesis. PMID- 12738526 TI - Endometrial cyst of the liver: case report and review. AB - OBJECTIVE: To describe a patient with a large endometrial cyst of the liver and review the literature concerning these rare lesions. DESIGN: Case report. SETTING: University hospital. PATIENT(S): A 42-year-old nulligravid woman with a 22 x 24 x 30 cm cyst in the right lobe of the liver. INTERVENTION(S): Thorough exploration of the abdominal cavity revealed no evidence of endometrial implants elsewhere. A complete excision of the cyst was performed. MAIN OUTCOME MEASURE(S): Surgical specimen. RESULT(S): Histologic examination of the surgical specimen confirmed the diagnosis of endometriosis. CONCLUSION(S): Endometriosis should be included in the differential diagnosis list of a peripherally located cystic liver mass in women. PMID- 12738527 TI - Oocyte collection during cryopreservation of the ovarian cortex. PMID- 12738528 TI - Hemodynamic effect of danazol therapy in women with uterine leiomyomata. PMID- 12738530 TI - Expression and presence of the platelet-activating factor receptor in human embryos. PMID- 12738529 TI - Cigarette smoking and the male-female sex ratio. PMID- 12738531 TI - Betadine (povidone-iodine) is toxic to murine embryogenesis. PMID- 12738533 TI - Predictive values of transvaginal sonography in specifying bicornuate and septate uterus? PMID- 12738532 TI - Effect of cotreatment with growth hormone on ovarian stimulation in poor responders to in vitro fertilization. PMID- 12738534 TI - Vasa previa, multiple pregnancies, and in vitro fertilization clarification. PMID- 12738536 TI - Cryopreservation techniques. PMID- 12738538 TI - Multiple confounders--measured with error? PMID- 12738540 TI - Heroic years. PMID- 12738541 TI - Heroic years. PMID- 12738543 TI - Wistar rat palpebral conjunctiva contains more slow-cycling stem cells that have larger proliferative capacity: implication for conjunctival epithelial homeostasis. AB - PURPOSE: To determine the location of conjunctival epithelial stem cells. METHODS: Wistar rats received daily injection of 5-bromo-2-deoxyuridine (BrdU) at a dose of 5 mg/100 g for 2 weeks followed by a 1-month BrdU-free period before death. After the rats were sacrificed, the orbital contents and eyelids were exenterated en bloc, fixed in buffer formalin, and embedded in paraffin. To compare the proliferative capacity of ocular epithelial cells, 1.0% phorbol myristate (TPA) in petrolatum was topically applied once daily to both eyes of Wistar rats for 12 days. After 6, 12, 18, and 24 hours and 2, 4, 6, 8, 10, and 12 days of TPA treatment, rats were administered BrdU intraperitoneally 7 hours before they were sacrificed. The ocular epithelium was fixed and processed for immunochemistry, and the labeling index (LI) of every epithelial zone was determined. RESULTS: Slow-cycling cells, detected as label-retaining cells (LRCs), were found in bulbar, fornical, and palpebral epithelia and mucocutaneous junctions, as well as in limbal epithelia. The greatest numbers of LRCs were identified in palpebral epithelium. Under normal situations, in conjunctiva the LI was lowest in palpebral epithelium (2.1 +/- 0.5) compared with bulbar (2.2 +/- 0.5), fornical (2.3 +/- 0.4) epithelia and mucocutaneous junction (3.4 +/- 0.9), respectively. In cornea, the LI was lowest in limbal epithelium (1.8 +/- 0.7) compared with central corneal epithelium (3.5 +/- 0.6). Twenty-four hours after TPA treatment, an 8.2-fold increase in the palpebral epithelial basal cell labeling index was noted compared with 4.7-fold, 5.7-fold, and 3.8-fold increases in bulbar, fornical, and mucocutaneous junction epithelial basal cell labeling indices, and a sevenfold increase in the limbal basal cell labeling indices compared with a 2.1-fold increase in the corneal basal cell labeling index, respectively. Limbal and palpebral epithelia maintained a significantly greater proliferative response (5.5-to 6.3-fold increase, respectively) during chronic stimulation than corneal, bulbar, fornical epithelia, and mucocutaneous junction (0.6- to 2.3-fold increase, respectively). CONCLUSIONS: In Wistar rat conjunctiva, slow-cycling cells are primarily located in palpebral epithelium, which has greater proliferative capacity than other conjunctival epithelia. This finding means that, in the Wistar rat, the conjunctival epithelial stem cells are mainly located in palpebral epithelium. These data open new perspectives in ocular epithelial development and are relevant in conjunctival wound repair. PMID- 12738544 TI - Influence of dorzolamide on corneal endothelium. AB - PURPOSE: To evaluate the influence of topical 1% dorzolamide treatment for 3 months on the endothelial morphology and the thickness of the cornea. METHODS: The corneal endothelium was investigated in 21 glaucoma patients (29 eyes) who received topical 1% dorzolamide treatment for 3 months. Intraocular pressure, corneal endothelium, and central corneal thickness (CCT) were measured before and 3 months after topical dorzolamide treatment. The relation between the endothelial cell density loss or the CCT increase, and factors such as the history of previous intraocular surgery, the presence of type II diabetes mellitus, type of glaucoma, intraocular pressure before treatment (above or below 20 mm Hg), or the kinds of eye drops used before dorzolamide treatment were evaluated. RESULTS: The endothelial cell density, coefficient of variation of cell area, and percentage of hexagonal cells did not change significantly. The CCT after treatment (537 +/- 34 microm) was significantly increased over that before treatment (530 +/- 33 microm). There was no significant relation of endothelial cell loss or CCT increase to any of the above factors. CONCLUSIONS: Topical 1% dorzolamide treatment increases CCT and does not affect corneal endothelial morphology. PMID- 12738545 TI - Expression of functional ICAM-1 on cultured human keratocytes induced by tumor necrosis factor-alpha. AB - PURPOSE: Leukocytes such as neutrophils contribute to the pathogenesis of corneal ulcer. The effect of the proinflammatory cytokine tumor necrosis factor (TNF) alpha on the expression of intercellular adhesion molecule (ICAM)-1 by cultured human keratocytes was investigated because the interaction of leukocytes with ICAM-1 expressed on the surface of structural cells mediates leukocyte infiltration into tissue at sites of inflammation. METHODS: Cultured human keratocytes were incubated with various concentrations of TNF-alpha. The surface expression of ICAM-1 was evaluated by whole-cell enzyme-linked immunosorbent assay, flow cytometry, and immunohistochemistry. The abundance of ICAM-1 mRNA in cell lysate was determined by quantitative reverse transcription and polymerase chain reaction analysis. Adhesion of neutrophils to corneal fibroblasts was assayed by measuring the fluorescence of Calcein-AM-labeled neutrophils. RESULTS: Incubation of keratocytes with TNF-alpha induced increased expression of ICAM-1 on the surface of keratocytes in a dose- and time-dependent manner. The abundance of ICAM-1 mRNA in keratocytes was increased by the incubation of cells with TNF alpha. Exposure of keratocytes to TNF-alpha increased the adherence of human neutrophils to these cells. CONCLUSIONS: Stimulation of keratocytes with TNF alpha resulted in an increase in the abundance of ICAM-1 mRNA, the cell surface expression of ICAM-1 protein, and enhanced adhesion of neutrophils to these cells. The expression of ICAM-1 on human keratocytes may thus contribute to leukocyte infiltration into the corneal stroma of individuals with inflammatory ocular diseases. PMID- 12738546 TI - Falsely elevated intraocular pressure due to an abnormally thick cornea in a patient with nevus of Ota. AB - BACKGROUND: Several ocular complications, including glaucoma and ocular hypertension, have been reported in patients with nevus of Ota. CASE: A 12-year old boy with nevus of Ota on the left side of his face was referred for further examination of elevated intraocular pressure in his left eye. OBSERVATIONS: Intraocular pressure measured with Goldmann tonometry was 19 mm Hg OD and 25 mm Hg OS. No visual field defects were detected by Goldmann or Humphrey perimetry. With ultrasonic pachymetry, the mean +/- SD of central corneal thickness was 560 +/- 4 microm in the right eye and 652 +/- 9 microm in the left. CONCLUSION: Ophthalmologists should be aware that the abnormally thick cornea of patients with nevus of Ota may produce falsely elevated intraocular pressure readings. PMID- 12738547 TI - Passive length-tensile properties of extraocular muscles under botulinum toxin type C. AB - PURPOSE: Passive length-tension of the extraocular muscle was measured after injection of botulinum neurotoxin type C (BoNT-C) to evaluate its chemo denervation effect. METHODS: BoNT-C was injected unilaterally at 5.0 or 2.5 units in the superior rectus muscle of the albino rabbit. The muscle was separated several times at its original insertion between 3 days to 8 weeks after injection, and the passive length-tension produced by stretching the muscle from its physiologic length was measured. The length-tension curve was analyzed, and the passive load was determined from early components. The compliance was determined by approximating the entire ascending curve by an exponential function. RESULTS: In the 5.0-unit group, the passive load increased significantly 2 weeks after injection but decreased to the level of the control group after 4 weeks and remained at that level until after 8 weeks after injection. In the 2.5-unit group, also, there were changes similar to those in the 5.0-unit group, but the changes were not significant compared to the control group. No significant change was observed in the compliance in either group. CONCLUSIONS: Persistent chemo-denervation effect was not observed in passive length-tension at the doses of BoNT-C used in this study. The increase in the passive load soon after injection was suggested to have been caused by the same mechanism as BoNT-A. PMID- 12738548 TI - Histopathological study of a case with glaucoma due to Sturge-Weber syndrome. AB - PURPOSE: To investigate the cause of the secondary glaucoma in a case of Sturge Weber syndrome by histopathology. CASE: A 10-year-old boy with Sturge-Weber syndrome and glaucoma in the right eye was studied. Trabeculectomy was performed because of uncontrolled intraocular pressure, and the trabeculectomy specimen was examined histologically by both light and electron microscopy. RESULTS: Histological examination of the trabeculectomy specimen showed that the ciliary muscle was dislocated anteriorly, and the Schlemm canal was not present. The spaces in the juxtacanalicular connective tissue (JCT) were replaced by vascular structures and connective tissue. There were two kinds of vascular structures: in one, the endothelium was surrounded by pericytes; and in the other, the endothelium was not surrounded by pericytes. CONCLUSIONS: Developmental abnormalities of the Schlemm canal and the JCT may have caused the glaucoma. These observations suggest that the developmental abnormalities of both the mesoderm and the neural crest might be involved in the pathogenesis of the glaucoma in cases of Sturge-Weber syndrome. PMID- 12738549 TI - Platelet-derived growth factor receptor kinase inhibitor AG1295 and inhibition of experimental proliferative vitreoretinopathy. AB - PURPOSE: Receptor tyrosine kinase (RTK) activation is critical for growth factor mediated cell proliferation. The present study was designed to determine the effect of tyrphostin AG1295, a selective blocker of platelet-derived growth factor (PDGF) RTK, on proliferative vitreoretinopathy (PVR) development. METHODS: Rabbit conjunctival fibroblasts cells (1 x 10(4)) were seeded into 96-well plates and maintained in Dulbecco's modified essentialmedium (DMEM) with 0.5% fetal bovine serum. The cells were exposed to 50 ng/mL PDGF-AAor PDGF-BBor phosphate buffered saline with or without AG1295 (1 microM, 10 microM, and 100 microM). After 3 days, the viable cells in each well were measured by 3,(4,5-dimethyl-2 thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Homologous rabbit conjunctival fibroblasts were injected intravitreally, followed by injection of 100 microM of AG1295. The development of tractional retinal detachment (TRD) was assessed to evaluate the effect of AG1295 in vivo. Electroretinography and histologic studies were performed after intravitreal injection of AG1295 into untreated eyes to evaluate toxicity. RESULTS: Two concentrations of AG1295 (10 and 100 microM) significantly inhibited rabbit conjunctival fibroblast cell growth stimulated by PDGF-AA or PDGF-BB in vitro. Development of TRD was significantly attenuated (P <.01) with 100 microM of AG1295 until day 21. No significant histologic or retinal functional damage was found in the AG1295 treated group. CONCLUSIONS: PDGF receptor specific inhibitor AG1295 attenuated PVR without significant side effects in rabbits. This reagent could be a useful treatment to prevent PVR. PMID- 12738550 TI - Analysis of cytokine mRNAs in murine herpes simplex virus type 1 retinitis. AB - PURPOSE: To investigate cytokine mRNA expression during the inflammatory process induced in the contralateral eyes by uniocular inoculation of herpes simplex virus type 1 (HSV-1) via the anterior chamber. METHODS: BALB/c mice were inoculated in the anterior chamber with 5 x 10(4) plaque-forming units of HSV-1 (KOS). mRNA was extracted from the inflamed posterior segments of the uninoculated eyes at 0 (control), 9, 11, 14, and 21 days postinoculation (p.i.). Reverse transcription-polymerase chain reaction was performed for semiquantitative analysis of mRNA expression of interleukin (IL)-1beta, IL-2, IL 4, IL-10, IL-12p35, IL-12p40, interferon (IFN)gamma, tumor necrosis factor (TNF)alpha, transforming growth factor (TGF)beta2 and induced nitric oxide synthase (iNOS). RESULTS: Peak mRNA expression of iNOS was observed at day 14 p.i. The time profiles of mRNA expression for IL-1beta, IL-2, IL-4, IL-10, IFN gamma, TNFalpha were similar to that of iNOS, while TGFbeta2, IL-12p35, and IL 12p40 demonstrated a reverse pattern. CONCLUSIONS: The kinetics of the analyzed cytokines synchronized with the clinicopathological activity of the experimental murine HSV-1 retinitis. The immunosuppressive cytokines TGFbeta2 and IL-10 demonstrated different peaks of mRNA expressions suggesting that the down regulation phase of the inflammatory process was controlled by several factors working at different phases. PMID- 12738551 TI - Reproducibility of the measurements of the optic nerve head topographic variables with a confocal scanning laser ophthalmoscope. AB - PURPOSE: To determine the optic nerve head topographic parameters with the least variability in repeat measurements. METHODS: We randomly selected and evaluated 1 eye each of 20 healthy subjects, a total of 20 eyes. We used the confocal scanning laser ophthalmoscope (CSLO) for optic nerve head analysis. The disc area and a total of 13 parameters were determined by Top SS. Each subject was examined five times, each time on a different day, during a 2-month period. We obtained a series of five 10 x 10 degrees images for each eye per visit. Three of the five images were randomly selected to create a mean image. Coefficients of variation of each of the 13 variables studied were calculated separately by using those five different optic nerve head topographic measurements. RESULTS: The subjects were 11 women and 9 men. The mean age of the subjects was 30.5 +/- 6.9. The mean optic disc areas were 2.26 +/- 0.39 mm(2) and 1.96 +/- 0.37 mm(2) for the men and the women, respectively (P >.05). The mean coefficient of variation for measurement of the variables was found to range between 1.2% and 9.8%. The variables, cup shape, volume above, average depth, and volume below, were found to yield the best reproducible measurements. CONCLUSION: The CSLO with its highly reproducible measurements (<10% error) offers a highly objective, safe, and effective method for clinical use in measurements of the topography of the optic nerve head. The subtle differences in the variables, cup shape, volume above, average depth, and volume below, may be of vital importance in the follow-up of those diseases requiring longitudinal monitoring of the optic nerve head, namely ocular hypertension and glaucoma. PMID- 12738552 TI - Alternative method of evaluating visual field deterioration in very advanced glaucomatous eye by microperimetry. AB - PURPOSE: To report an alternative method of evaluating visual field deterioration in very advanced glaucomatous eyes. METHODS: This prospective, comparative study included 24 eyes of 24 consecutive patients with very advanced glaucoma. The eyes were divided into two groups based on their visual field measurements by Goldmann perimetry: Group 1 had a central island with or without a detached temporal island (n = 13), and Group 2 had a central island with a temporal island connected by only a thin isthmus (n = 11). The size of the central island was determined by Goldmann perimetry, and microperimetry plots were determined around the disc margin. The angle formed between a line passing from the disc center to the "seen" points and a second line passing to the "not seen" points at the superior temporal or the inferior temporal sites was used for the analyses. RESULTS: The mean size of the island was 6.3 +/- 1.9 in Group 1 and was 7.0 +/- 2.0 in Group 2 (P =.38). The mean angle between the "seen" and "not seen" points in Groups 1 and 2 was 56.1 +/- 6.9 and 76.1 +/- 11.8, respectively (P =.0002). The reduction in the angle correlated with the visual field loss (P =.018), but the reduction in the size of the central island was not significantly associated with the visual field loss (P =.29). CONCLUSION: Microperimetry may be an alternative method of evaluating advanced glaucoma. PMID- 12738553 TI - Adenosine A1 and benzodiazepine receptors and glucose metabolism in the visual structures of rats monocularly deprived by enucleation or eyelid suture at a sensitive period. AB - PURPOSE: To determine the changes in the adenosine A(1) and benzodiazepine receptor density and in glucose metabolism in the visual centers of the rat brain following monocular enucleation or eyelid suture on postnatal day 10 (PN10). METHODS: Following monocular enucleation or eyelid suture on PN10 rats, the alterations in adenosine A(1) and benzodiazepine receptor density, and in glucose metabolism were evaluated in the superior colliculus (SC), the dorsal lateral geniculate body (DLG), and the visual cortex (VC) by ex vivo autoradiography with [11C]MPDX, [11C]flumazenil and [14C]2-deoxyglucose, respectively. RESULTS: Enucleation reduced the [11C]MPDX binding in the SC and DLG, and enhanced the [11C]flumazenil binding in the SC. Eyelid suture reduced the [11C]flumazenil binding in the VC at day 20. [14C]2-deoxyglucose uptake was not decreased by enucleation in any region except in the SC and DLG at day 1, but was decreased by eyelid suture in the SC at days 20 and 55 and in the VC at day 55. CONCLUSIONS: The decrease in the presynaptic adenosine A(1) receptors in the SC following enucleation is coupled with an upregulation of postsynaptic benzodiazepine receptors. These neural reactions are completely different from those following eyelid suture. The development of neural architecture for visual functions is not completed at PN10 in rats. PMID- 12738554 TI - Association of life-style with intraocular pressure in middle-aged and older Japanese residents. AB - PURPOSE: To evaluate the associations of several life-style-related factors with intraocular pressure (IOP). METHODS: Of a total of 649 men and women who had been examined in an annual health check-up being conducted by a general hospital in Ibaraki prefecture, Japan, 569 individuals (age range, 29-79 years) who had not undergone any medical treatment for hypertension, ocular hypertension, or glaucoma, were selected as the subjects of this study. The associations of several life-style-related factors with the IOP were evaluated by multiple regression analyses and analyses of covariance. RESULTS: Body mass index (BMI), alcohol consumption score, and cigarette consumption were found to have a significantly positive association with the IOP in men (P for trend =.002, <.001, and <.001, respectively). In women also, the BMI was positively related to the IOP (P for trend =.071). In respect to the effects of coffee consumption, it was shown that in men the mean IOP adjusted for age, the BMI, alcohol intake score, cigarette consumption, and blood pressure were significantly lower in habitual coffee drinkers than in coffee abstainers (P =.016). CONCLUSION: The results of this study suggest that the IOP level may be substantially affected by daily life style among Japanese. PMID- 12738556 TI - Clinical features of comitant strabismus related to family history of strabismus or abnormalities in pregnancy and delivery. AB - PURPOSE: To elucidate the genetic or environmental background for clinical features in the three major types of comitant strabismus. METHODS: Interview based on a questionnaire asking background factors such as family history of strabismus and abnormalities in pregnancy and delivery was conducted with 101 consecutive patients with infantile esotropia (5-180 months of age), 83 with accommodative or partially accommodative esotropia (6-201 months of age), and 143 with intermittent exotropia (3-216 months of age) seen during 7 months from May to November 1998. The clinical features of strabismus obtained from medical records were analyzed statistically by logistic regression to determine their relation with these background factors. RESULTS: In infantile esotropia, patients with family history of strabismus had a significantly higher chance of showing latent nystagmus (odds ratio, 3.553; 95% confidence interval [CI], 1.077-11.717; P =.0373, logistic regression analysis). In a subgroup of 40 patients with infantile esotropia whose birth followed no abnormalities in pregnancy or delivery, patients with family history of strabismus had a significantly higher chance of showing inferior oblique muscle overaction (odds ratio, 7.714; 95% CI, 1.246-47.761; P =.0280), dissociated vertical deviation (odds ratio, 6.667; 95% CI, 1.176-37.787; P =.0321), and latent nystagmus (odds ratio, 7.333; 95% CI, 1.168-46.060; P =.0336). In accommodative or partially accommodative esotropia and intermittent exotropia, no relation was found between the clinical features and the background factors. CONCLUSIONS: Inferior oblique muscle overaction, dissociated vertical deviation, and latent nystagmus in infantile esotropia might have a genetic background. PMID- 12738555 TI - Influence of topical betaxolol and timolol on visual field in Japanese open-angle glaucoma patients. AB - PURPOSE: To assess the effects of topical betaxolol and timolol on the visual field in Japanese open-angle glaucoma (OAG) patients. METHODS: This study was a multicenter, 2-year, prospective, randomized and double-masked study. Tests using the Humphrey 30-2 perimeter program were conducted every 6 months and the data of 95 patients were analyzed using regression analysis. Estimated regression coefficients for mean deviation (MD), corrected pattern standard deviation (CPSD), and total deviation (TD) values clustered into 15 sectors were obtained for each treatment group. RESULTS: Estimated slopes (dB/year) for MD and CPSD showed no significant difference from zero in either group. However, in the betaxolol group, estimated slopes (dB/year) for two adjacent sectors in the inferior arcuate area were significantly positive (P =.0135,.0116) while in the timolol group, no significant difference from zero was seen in any of the sectors. IOP changes from baseline in the timolol group were greater than in the betaxolol group, although no statistical significance was seen at any of the examination times. CONCLUSION: MD and CPSD showed no significant change in either group. In the betaxolol group, however, a significant trend in improvement of visual field performance was seen in the inferior arcuate subfield. Timolol reduced IOP more effectively than betaxolol in OAG patients. PMID- 12738557 TI - Comparison of optic disc topography measured by Retinal Thickness Analyzer with measurement by Heidelberg Retina Tomograph II. AB - PURPOSE: To compare the optic disc topography measurements from a digitized laser slit-lamp and a confocal scanning laser ophthalmoscope (SLO). METHODS: Ten normal subjects (10 eyes) were recruited. Topographic measurements of the optic disc were performed three times in all eyes using the Retinal Thickness Analyzer (RTA) and the Heidelberg Retina Tomograph II (HRT II), and the mean values determined. The mean values of 11 optic disc parameters were compared between the two instruments by the Wilcoxon signed rank test. To test the reproducibility of the topographic measurements, the coefficients of variation (CV) of the topographic parameters among the three measurements from each of the two instruments were compared using the Wilcoxon signed rank test. RESULTS: The mean cup depth, mean retinal nerve fiber layer (RNFL) thickness, and RNFL cross-sectional area were significantly smaller when measured by RTA compared with HRT II (P =.0067, P =.0364, P =.0467, respectively). The mean CVs from the RTA were larger than those from the HRT II for all parameters; however, the differences in all parameters did not reach significance. CONCLUSION: The RTA measured smaller z-axis values compared with the HRT II. The reproducibility of the topographic data was not significantly different between the instruments. PMID- 12738558 TI - Japan's share of published research in ophthalmology. AB - PURPOSE: This study was conducted to determine Japan's share of published research in ophthalmology during the last decade. METHODS: Ophthalmology journals with higher impact factors were accessed through the Medline database to elicit the number of articles published in 1991-2000 that originated in various countries, including Japan. The proportion of articles with a higher grade of evidence (randomized controlled trials/clinical trials, cohort studies, and case control studies) was determined for Japan and compared with the average values of the total articles published in these journals. In addition, the percentage of published research from the 20 top-ranking countries was calculated, showing the trend over time. RESULTS: Of the total articles (21,327), Japan's share in the selected ophthalmology journals was 6.5% (1,387 articles), ranking third in the world, following the USA (51.5%) and the UK (11.3%). The recent increase in the share was statistically significant for Japan (P =.01). However, the proportionate value of clinical research evidence was lower for Japan-originated articles than the average value for the total clinical research articles in these journals. CONCLUSIONS: Appropriate measures should be taken in the ophthalmology field in Japan to increase the number of clinical research papers with a higher grade of evidence. PMID- 12738572 TI - Ciliochoroidal effusion syndrome secondary to Sturge-Weber syndrome. PMID- 12738574 TI - The experiences of migrant health nurses employed in seasonal satellite nurse managed centers: a qualitative study. AB - This research study describes the unique experiences of nurses who are employed in migrant health seasonal satellite nurse-managed centers in the upper Midwest. Data were generated through semistructured interviews with 10 seasonal nurses. Phenomenology served as the research method. Four themes were identified including seeking seasonal employment, establishing migrant seasonal satellite nurse-managed centers, learning the culture of Hispanic migrant farmworkers, and referring Hispanic migrant farmworkers for medical care. During their seasonal employment, nurses learned to establish and operate satellite nurse-managed centers. Due to the migrant health nurses' daily contact with their clients, they were able to establish rapport that led to a trusting relationship. This enabled them to provide culturally sensitive and lifestyle appropriate care to the migrant farmworker population. PMID- 12738575 TI - Health concerns and learning styles of underserved and uninsured clients at a nurse managed center. AB - The medically underserved and uninsured often seek care for immediate health concerns rather than use health promotion activities. To facilitate health promotion activities, it is important to understand the health concerns and learning styles of these underserved clients. The purpose of this descriptive study was to assess specific health concerns and identify learning styles of clients seeking care at a nurse managed center. Eighty-two participants completed the Hilltop Health Questionnaire, a self-report instrument developed by the research team. Top reasons for seeking care included upper respiratory infections (17%), routine physical (17%), medication refill (12%), and low back pain (11%). Preference for learning style included hands-on instruction (82%), reading (63%), and listening (59%). These data were used to obtain or customize teaching modules based on the clients' health concerns and preferred style of learning. Continued emphasis should be placed on health education that engages the underserved and uninsured in their care and health promotion activities. PMID- 12738576 TI - Promoting participation: evaluation of a health promotion program for low income seniors. AB - This article describes a qualitative evaluation of the Seniors Active Living in Vulnerable Elders (ALIVE) program, a 10-month health promotion program for low income seniors. Program interventions delivered in seniors' apartment buildings included exercise classes, health information sessions (i.e., health corners), and newsletters. The evaluation examined program participation, program impacts, and how the program worked. The most frequent reason for joining the program was recognizing the benefits of exercise, and the most frequent reason for not attending the program was having other priorities. The main participant impact was "feeling better." Specific impacts were also noted in physical, mental, and social domains. Fun, program delivery adaptations, autonomy, social interactions, and staff-participant relationships were discovered to be important program processes. These processes all contributed to participant's "comfort" in the program. How and why the program worked is examined in relation to Pender's (1996) revised health promotion model and implications for nursing are indicated. PMID- 12738577 TI - Measurement of cardiac functioning in older adults with heart failure after exercise. AB - Although the length of a hospital stay is very limited for patients with heart failure, many patients attempt to resume some activities of daily living soon after arrival at home. Home care nurses routinely assess symptoms indicative of recurrent heart failure and the need for hospital readmission. The purpose of this comparative study was to determine whether there was a significant difference in diastolic blood pressure and oxygen saturation after walking for 6 min. Twenty adults with heart failure in an outpatient cardiology clinic were evaluated after the 6-min walk test. Although diastolic blood pressure and oxygen saturation were not significantly different after walking for 6 min, functional status was significantly related to oxygen saturation. Further study with home care clients is needed to evaluate the relation between activity and oxygen saturation. PMID- 12738578 TI - Multipurpose senior centers: opportunities for community health nursing. AB - Nationally, almost 10 million older Americans are served by approximately 12,000 multipurpose senior centers (MSCs). Among those over age 65, 15% attended an MSC in the previous year. We can expect that the number of older Americans attending MSCs will increase as our population ages. We conducted face-to-face interviews with 59 older adults at an MSC that provides programming to approximately 350 individuals aged 60 and older during the course of 1 year. We identified several areas in which senior center participants could benefit from the presence of community health nursing: loneliness and social support; diet and nutrition, oral health, and self-rated health and exercise. Community health nurses should play an integral role in MSCs by providing direct healthcare in the form of interventions and referrals and by providing preventative care and advice so that senior center participants may maintain and improve the prospects for their independence and quality of life. PMID- 12738579 TI - Effects of environmental enrichment for mice: variation in experimental results. AB - This study focused on the effects of different enriched environments for mice in a number of behavioral and physiological parameters in 2 routine laboratory testing procedures: potency testing for tetanus vaccine and stress-induced hyperthermia. The variability in the results was studied by calculating and analyzing mean absolute devi-ations. Mice from enriched conditions weighed more and consumed more food than mice from standard housing conditions. However, mice from enriched conditions lost more body weight after being housed individually. Other physiological parameters showed no differences. Mice from standard conditions were more active in an open field, suggesting a tendency to overrespond to various stimuli in a testing environ-ment. Mice from enriched environments were more tranquil and easier to handle. The enrichment did not influence the variability in any of the parameters measured, al-though earlier results and results of other studies suggest that the effects on the vari-ability in results are parameter dependent. When enrichment does not influence vari ability, there is no reason for not introducing cage enrichment and by doing so contributing to the animals' welfare. PMID- 12738580 TI - Enclosure design and space utilization by Indian leopards (Panthera pardus) in four zoos in Southern India. AB - Enclosure design and the use of enclosure space influence the activity budget of cap-tive leopards. The study laid out in grids all enclosures on the base map and segregated these grids into 4 zones. Every 5 min, the study recorded the proportion of time spent in these zones with the leopards' behavior. Captive leopards most frequently used the "edge" zone. Almost all leopards used the edge zone for stereotypic pacing, the "back" zone for resting, and the "other" zone for activity. The study positively corre-lated the proportion of time spent in the "enriched" zone with activity levels exhibited by leopards housed in some enclosures and with resting in others. Thus, the study seg-regated structural objects in the enriched zone into activity-related features (e.g., logs) and rest related features (e.g., trees and sleeping platforms). Compared with individu-als housed in barren enclosures, leopards housed in structurally enriched on-exhibit enclosures exhibited higher levels of activity. Enclosure design was found to be an important factor influencing the welfare of leopards in captivity. PMID- 12738582 TI - Cage use and feeding height preferences of captive common marmosets (Callithrix j. jacchus) in two-tier cages. AB - Determining appropriate feeding regimes has important welfare implications for cap-tive primates. This study examined the preference of food bowl heights in 6 pairs of common marmosets (Callithrix jacchus) housed in a 2-tier cage system. Given that marmosets are arboreal and spend most of their time in the upper half of their cages, we predicted that the marmosets would prefer a food bowl positioned at the top of the cage over one positioned at the bottom. We further predicted that this would be more apparent for the marmosets housed in lower tier than upper tier cages. Given a choice regarding where to feed, marmosets did prefer the top bowl to the bottom bowl; how-ever, when only 1 food bowl was presented, its position had no significant effect on the marmosets' feeding behavior. In addition, contrary to the prediction, there were few differences in the marmosets' feeding behavior in the upper and lower tier cages. Feeding the marmosets in a bowl at the bottom of their cage did not result in greater cage use. On the basis of this study, we recommend positioning captive marmosets' food bowls high in the cage. PMID- 12738581 TI - Retreat space and human visitor density moderate undesirable behavior in petting zoo animals. AB - This study focused on the relationship between nonhuman animal behavior and envi ronment- specifically, between the undesirable behavior exhibited by domestic pet ting zoo animals in the presence of humans and the spatial design of the petting zoo environment. A spatial feature of a petting zoo referred to as a retreat space was ma-nipulated so that it affected the animals' opportunity for individual control over inter-action with humans. Three conditions were tested: no retreat space, semi-retreat space, and a full-retreat space. The subjects of this study were 5 African pygmy goats (Capra hircus) and 2 Romanov sheep (Ovis aries). Investigators used a focal sam-pling technique to analyze approximately 27 hr of behavioral data collected. The data were analyzed using multiple linear regression methods. The findings suggest that the full-retreat design beneficially moderated both sheep and goat behavior: Undesirable behaviors were lowest in the full-retreat condition. This study provides information that may improve human-animal interactions in a petting zoo setting and may in-crease animal well-being through exhibit design and management techniques. PMID- 12738583 TI - Artificial weaning of Old World monkeys: benefits and costs. AB - Permanent mother-infant separation prior to natural weaning is a common hus bandry practice in monkey breeding colonies. In the United States, all eight Re gional Primate Research Centers have such colonies. Under undisturbed conditions, Old World monkey mothers wean their infants at the age of about 1 year (Hall & DeVore, 1965; Poirier, 1970; Roonwal & Mohnot, 1977; Southwick, Beg, & Siddiqi, 1965). Natural weaning is a gradual process. It implies that the mother, over a period of several weeks or months, more and more consistently discourages her infant to suck on her breasts. Once the mother stops nursing the infant for good, the affectionate bond between the two is not broken (Altmann, Altmann, Hausfater, & McCuskey, 1977; Lindburg, 1971; Poirier, 1970; Roonwal & Mohnot, 1977). The young usually remains in the ma-ternal group at least until prepuberty. Under confinement conditions, artificial weaning is an abrupt occurrence that takes place several months prior to the biologically normal age of weaning. It im-plies that the still-nursed infant is taken away from the mother and subsequently reared alone or with other artificially weaned infants. PMID- 12738584 TI - Animal dissection and evidence-based life-science and health-professions education. AB - Cambridge mathematician and philosopher W. K. Clifford (1879/1999) con-cluded his famous essay, "The Ethics of Belief" with the bold claim that "it is wrong always, everywhere, and for anyone to believe anything upon insufficient evidence" (p.77). Clifford's enthusiasm for evidentialism-the principle that one should proportion one's belief to the strength of the evidence-may have been overzealous, but a plausible interpretation of his view is this: Because beliefs of-ten have serious moral consequences, one should base one's beliefs on the evi dence, and it is intellectually and morally irresponsible not to do so. This per spective motivates recent so-called "evidence-based" methods in the fields of medicine and education. Balcombe's (2000, 2001) case for replacing learning methods that require pain, suffering, and death for animals with methods that do not (computer-assisted learning, three-dimensional models, videotapes, and other alternatives) can be seen as motivated by this evidentialist perspective. Balcombe provided a wealth of empirical evidence from educational studies to show that in most contexts animal dissection is not necessary-and even counterproductive-to achieve valid educa-tional goals, especially higher order goals (concept learning and problem solving). He demonstrated that no sound defense of dissection has been given. Can we learn as effectively without hurting or killing another being? If so, why do we not try? Many of the studies Balcombe cites have supported sufficiently the adequacy and, often, superiority of learning methods that do not harm animals or students. The first of the aforementioned questions is being answered; we can learn effectively with these non-detrimental methods. Those who seek to educate (and accept the prin-ciple of "do no harm") must seize the second question because they see, in the big pic ture, the benefit for themselves, their students, their society, and other sentient beings. (p. 132) PMID- 12738585 TI - Ethics and animal welfare evaluations in South East Asian zoos: a case study of Thailand. AB - Concern for zoo animals is palpable throughout society in many countries in South East Asia. It is important to understand problems of animal welfare in order for zoos to make significant improvement in maintaining high standards. With a case study of 3 zoos in Thailand, this article presents results for the first time on how ethics and welfare evaluations are conducted in South East Asian zoos. The study identified several major and minor welfare problems and provided constructive suggestions to zoo authorities, which in turn significantly improved the standards of animal welfare. Thus, the data presented in this article could serve as a model for other zoos to follow animal welfare evaluations locally, regionally, and globally. PMID- 12738586 TI - Implementation of a feral cat management program on a university campus. AB - In August 1998, Texas AM University implemented on campus a trap-test-vaccinate alter-return-monitor (TTVARM) program to manage the feral cat population. TTVARM is an internationally recognized term for trapping and neutering programs aimed at management of feral cat populations. In this article we summarize results of the program for the period August 1998 to July 2000. In surgery laboratories, senior veterinary students examined cats that were humanely trapped once a month and tested them for feline leukemia and feline immunodeficiency virus infections, vaccinated, and surgically neutered them. They euthanized cats testing positive for either infectious disease. Volunteers provided food and observed the cats that were returned to their capture sites on campus and maintained in managed colonies. The program placed kittens and tame cats for adoption; cats totaled 158. Of the majority of 158 captured cats, there were less kittens caught in Year 2 than in Year 1. The proportion of tame cats trapped was significantly greater in Year 2 than in Year 1. The prevalence found for feline leukemia and feline immunodeficiency virus ELISA test positives was 5.8% and 6.5%, respectively. Following surgery, 101 cats returned to campus. The project recaptured, retested, and revaccinated more than one-fourth of the cats due for their annual vaccinations. The program placed 32 kittens, juveniles, and tame adults for adoption. The number of cat complaints received by the university's pest control service decreased from Year 1 to Year 2. PMID- 12738587 TI - Prediction of adoption versus euthanasia among dogs and cats in a California animal shelter. AB - The purpose of this retrospective cohort study was to investigate determinants of adoption of cats and dogs from a large municipal animal shelter. The subjects were 4,813 cats and 3,301 dogs impounded by the Sacramento County Department of Animal Care and Regulation and offered for adoption September 9, 1994 to May 26, 1995. The study constructed models predicting the conditional probability of adoption using logistic regression and a final multiple logistic regression model from variables found to be important predictors of adoption. Age, sex, coat color, and reason for relinquishment were major determinants of adoption in cats. Age, sex, coat color, reason for relinquishment, breed, purebred status, and injury status were major determinants of adoption in dogs. Shelter personnel could utilize this information to increase the adoption of frequently overlooked animals. Alternatively, shelters could use this to focus their resources on animals with characteristics the public prefers. PMID- 12738588 TI - Selecting shelter dogs for service dog training. AB - Service dogs are an essential aid to persons with disabilities, providing independence, mobility, and improved self-esteem. Because of these proven benefits, the growing se of service dogs is creating a demand and supply crisis. One major cause is the 50% verage dropout rate for dogs selected for training. Weiss and Greenber (1997) re-cently found that a dog, successful on the most commonly used selection test items, was as likely to be either a poor or a good candidate for service work. The experiment presented here evaluated test items developed by the author in 15 years of experience with dogs. The test items were administered to 75 dogs from the Kansas Humane So-ciety. Once tested, the dogs received obedience and retrieval training. The experiment assessed each dog on behavior over 5 weeks of training versus performance on each selection test item. A subset of the selection items, combined in a regression analysis, accounted for 36.4% of the variance with R = 0.603. This research also revealed a reli-able test for dog aggression without risking injury to dog or tester. Items for testing in-cluded fear, motivation, and submission. Another set of selection items reliably pre-dicted the trait of "high energy" commonly described as "high strung." Future research should involve investigating the effectiveness of both cortisol levels and blood pressure in predicting traits to help strengthen the predictive value of the tool and then testing on dogs trained to be full service dogs. PMID- 12738589 TI - Cage size preference in rats in the laboratory. AB - The size of an enclosure is an integral part of how well it accommodates a nonhuman animal's welfare; however, most enrichment studies concentrate on modifying the area inside the enclosure rather than enlarging it. It has been suggested that rats have little need for more cage space, but there is no empirical evidence about rats' need for space. This experiment provides preliminary evidence for the preferences of 5 male and 5 female albino rats using T-maze choices followed by 5 min dwelling times. The rats showed a moderate but significant preference for the larger of 2 cages [540 cm2 vs. 1,620 cm2; binomial z, p <.05]. When the rats shared the chosen cage with 4 familiar cage mates, their preference for the larger cage did not become any stronger [paired t[9] = .820, p >.05]. The results suggest that rats should be given a somewhat larger space allowance but could share it with up to 4 other rats. PMID- 12738590 TI - The attending veterinarian's contract of employment: curse or blessing for animals in research institutions? AB - When attending veterinarians are not provided adequate job security by research in-stitutions, there is no guarantee that they are reliable allies of nonhuman animals and implement the provisions set forth in the federal animal welfare regulations. PMID- 12738591 TI - A study of three IACUCs and their views of scientific merit and alternatives. AB - Two ethical issues facing Institutional Animal Care and Use Committees [IACUCs] today are assessing scientific merit and the use of alternatives in research proposals. This study evaluated 3 IACUCs using a 19-question survey, with a 77.8% response rate. Although 76% of members answered that scientific merit should be more dili-gently assessed if more than slight pain is caused, 14% believed that assessing scien-tific merit is not the role of the IACUC. Nearly 86% agreed that the search for alterna-tives should be more diligent for protocols that incur more than slight pain to the animals involved. Some members believed that alternatives were not actively enough sought after, while others believed no viable alternatives existed. Additional guide-lines are needed to clarify these issues. PMID- 12738592 TI - Are we really reducing tidal volume--and should we? PMID- 12738593 TI - Role of isoniazid in once-weekly rifapentine treatment of pulmonary tuberculosis. PMID- 12738594 TI - The hunt for the elusive surrogate marker of sterilizing activity in tuberculosis treatment. PMID- 12738595 TI - Further discussion on breath condensate analysis. PMID- 12738596 TI - Perceptions, confessions: a systems response is needed. PMID- 12738597 TI - Discrepancies between perceptions by physicians and nursing staff of intensive care unit end-of-life decisions. AB - Several studies have pointed out ethical shortcomings in the decision-making process for withholding or withdrawing life-supporting treatments. We conducted a study to evaluate the perceptions of all caregivers involved in this process in the intensive care unit. A closed-ended questionnaire was completed by 3,156 nursing staff members and 521 physicians from 133 French intensive care units (participation rate, 42%). Decision-making processes were perceived as satisfactory by 73% of physicians and by only 33% of the nursing staff. More than 90% of caregivers believed that decision-making should be collaborative, but 50% of physicians and only 27% of nursing staff members believed that the nursing staff was actually involved (p < 0.001). Fear of litigation was a reason given by physicians for modifying information given to competent patients, families, and nursing staff. Perceptions by nursing staff may be a reliable indicator of the quality of medical decision-making processes and may serve as a simple and effective tool for evaluating everyday practice. Recommendations and legislation may help to build consensus and avoid conflicts among caregivers at each step of the decision-making process. PMID- 12738599 TI - Activated protein C inhibits the expression of platelet-derived growth factor in the lung. AB - The natural anticoagulant-activated protein C may inhibit inflammation and fibrosis in the lung. Platelet-derived growth factor is involved in the pathogenesis of lung fibrosis. This study assessed the effect of activated protein C on platelet-derived growth factor expression in human cell lines and in an in vivo model of lung fibrosis. Activated protein C significantly inhibited the secretion and expression of platelet-derived growth factor in human lung cell lines, primary bronchial epithelial cells, and macrophages. In vitro studies also showed that the endothelial activated protein C receptor is expressed by lung epithelial cells and macrophages, and that this receptor and the proteolytic activity of activated protein are implicated in the inhibition of platelet derived growth factor expression. In the in vivo model of lung fibrosis, intratracheal administration of activated protein C decreased the expression of platelet-derived growth factor and suppressed the development of lung fibrosis. Concomitant intratracheal administration of activated protein C and anti endothelial activated protein C receptor or anti-platelet-derived growth factor suppressed the inhibitory activity of activated protein C in vivo. In brief, this study describes a novel biological function of activated protein C that may further explain its inhibitory activity on lung inflammation and fibrosis. PMID- 12738598 TI - Wheezing, allergy, and parasite infection in children in urban and rural Ethiopia. AB - Epidemiological studies in developing countries suggest that intestinal parasite infection may reduce the risk of asthma. Because this evidence is all derived from adults and older children, we have investigated the relation between parasite infection, wheezing, and allergen skin sensitization in nested case control studies drawn from a survey of 7,155 children aged 1 to 4 years living in urban and rural areas of Jimma, Ethiopia. Infection with parasites was common, predominantly with Trichuris (54%), Ascaris (38%), and hookworm (10%). Wheezing in the past year was significantly more prevalent in urban (4.4%) than rural children (2.0%), and was less prevalent in those infected with Ascaris (age, sex, and urban/rural adjusted odds ratio, 0.5; 95% confidence interval, 0.3 to 0.9), particularly in relation to high-intensity infection. Similar, although nonsignificant, associations were found for hookworm (adjusted odds ratio, 0.6; 95% confidence interval, 0.2 to 1.8), but there was no suggestion of any relation to Trichuris infection. Dermatophagoides pteronyssinus and cockroach (Blattella germanica) skin sensitization was more prevalent in rural than urban children, and was unrelated to wheeze. We conclude that Ascaris and possibly hookworm infection protects against wheeze in young Ethiopian children, and that this effect is not mediated by inhibition of allergen sensitization. PMID- 12738600 TI - A decision rule for diagnostic testing in obstructive sleep apnea. AB - Obstructive sleep apnea (OSA) is traditionally diagnosed using overnight polysomnography. Decision rules may provide an alternative to polysomnography. A consecutive series of patients referred to a tertiary sleep center underwent prospective evaluation with the upper airway physical examination protocol, followed by determination of the respiratory disturbance index using a portable monitor. Seventy-five patients were evaluated with the upper airway physical examination protocol. Historic predictors included age, snoring, witnessed apneas, and hypertension. Physical examination-based predictors included body mass index, neck circumference, mandibular protrusion, thyro-rami distance, sterno-mental distance, sterno-mental displacement, thyro-mental displacement, cricomental space, pharyngeal grade, Sampsoon-Young classification, and over bite. A decision rule was developed using three predictors: a cricomental space of 1.5 cm or less, a pharyngeal grade of more than II, and the presence of overbite. In patients with all three predictors (17%), the decision rule had a positive predictive value of 95% (95% confidence interval [CI], 75-100%) and a negative predictive value of 49% (95% CI, 35-63%). A cricomental space of more than 1.5 cm (27% of patients) excluded OSA (negative predictive value of 100%, 95% CI, 75-100%). Comparable performance was obtained in a validation sample of 50 patients referred for diagnostic testing. This decision rule provides a simple, reliable, and accurate method of identifying a subset patients with, and perhaps more importantly, without OSA. PMID- 12738601 TI - Alveolar liquid clearance and sodium channel expression are decreased in transplanted canine lungs. AB - To determine the impact of transplantation-associated injury on the clearance mechanisms of pulmonary edema, we created a canine single lung transplant model. After 3 hours of preservation and 4 hours of reperfusion, right native lungs and left transplanted lungs were used to measure alveolar liquid clearance (ALC) in ex vivo liquid-filled lung preparations. We also examined the role of the pulmonary circulation in edema clearance in in vivo liquid-filled lungs between 4 and 8 hours of reperfusion. To study molecular modifications in ALC, we also measured expression levels of the epithelial sodium channel (ENaC) and sodium potassium-adenosine triphosphatase (ATPase). We found that ALC was significantly lower in transplanted than in right native lungs ex vivo (p < 0.05) and that transplanted lungs did not respond to the beta-adrenergic agonist terbutaline. Our in vivo study confirmed the ex vivo results. Molecular analyses revealed that ENaC messenger RNA but not sodium-potassium-ATPase was significantly decreased in transplanted lungs (p < 0.01). Furthermore, there was a significant decrease in ENaC protein expression. Therefore, we conclude that the current investigation indicates that the lung injury caused by lung preservation and transplantation significantly reduces the edema clearance ability of transplanted lungs. PMID- 12738602 TI - ATS/ACCP statement on cardiopulmonary exercise testing. PMID- 12738604 TI - Toll-like receptor-mediated tumor necrosis factor and interleukin-10 production differ during systemic inflammation. AB - Major trauma is associated with a decreased capacity of patients' leukocytes to produce proinflammatory cytokines on in vitro stimulation. We studied leukocytes from 48 patients with trauma and showed that this hyporeactivity was restricted to gram-negative bacteria, Escherichia coli endotoxin, and unmethylated bacterial DNA, whereas Leptospira interrogans endotoxin-induced tumor necrosis factor production was similar to that observed with healthy donors. As well, tumor necrosis factor and interleukin-6 production in response to gram-positive bacteria was not altered. The expression of toll-like receptor (TLR) 2 was not reduced on patients' monocytes as compared with healthy control subjects, whereas that of TLR4 was reduced. However, the hyporeactivity to gram-negative bacteria and E. coli endotoxin cannot be fully explained by the downregulation of TLR4. Indeed, unlike proinflammatory cytokines, after stimulation with these microbial products the release of antiinflammatory cytokines was increased as compared with healthy control subjects. The increased interleukin-10 production was analyzed in terms of intracellular signaling in peripheral blood mononuclear cells from patients with trauma: our results suggest the involvement of p38 mitogen activated protein kinase, Sp-1 transcription factor, heterotrimeric Gi protein, and phosphatidylinositol-3'-kinase. In conclusion, the immunodysregulation described for patients with trauma is not a generalized phenomenon but depends on the stimulus and the signaling pathway. PMID- 12738605 TI - Pulmonary dysfunction in transfusion-dependent patients with thalassemia major. AB - Pulmonary function tests were performed on 62 transfusion-dependent patients with thalassemia major, ranging in age from 8 to 33 years, and receiving chelation therapy with desferrioxamine or deferiprone. Percent predicted values for FVC, FEV1, and PEF were significantly reduced, whereas FEV1/FVC and maximal expiratory flow at 25% FVC were within normal limits, indicating a restrictive disease. Both FVC and FEV1 were negatively correlated with transfusional iron burden as indexed by age. Single-breath carbon monoxide transfer factor was reduced, even after correction for low hemoglobin concentration, and was negatively correlated with iron burden and iron overload, as indexed by serum ferritin levels. Owing to low hemoglobin concentration, blood-diffusing capacity was reduced, in spite of increased lung capillary blood volume, which was, however, adequate to normalize blood diffusing capacity when hemoglobin concentration was only partially restored by transfusion. The diffusing capacity of the alveolar-capillary membrane was substantially decreased and negatively correlated with age and serum ferritin, the fall being primarily attributed to increased membrane thickness. These findings suggest that lung fibrosis and/or interstitial edema related to iron overload are the main cause of pulmonary dysfunction observed in patients with thalassemia major. PMID- 12738606 TI - The Sarcoidosis Health Questionnaire: a new measure of health-related quality of life. AB - The Sarcoidosis Health Questionnaire (SHQ) is a health-related quality-of-life (HRQL) instrument we designed in a two-part study to allow patients to describe their satisfaction with life as it is affected by sarcoidosis. In the Development Study, we created the SHQ from a 151-item pool generated from interviews with 107 patients, a search of the relevant literature, and discussion with sarcoidosis experts. Using clinical impact methodology and the questionnaire responses of a separate group of 149 patients, we reduced the total number of items to 29. The final SHQ has three domains: Daily Functioning, Physical Functioning, and Emotional Functioning. In the Validation Study, performed with a different group of 111 patients, we found that the SHQ had good internal consistency reliability, as well as evidence of content, criterion, and construct validity based on its comparison with other measures of HRQL (Medical Outcomes Study 36-Item short form and the St. George's Respiratory Questionnaire) and of mental health (the Center for Epidemiologic Study-Depression Scale), as well as with clinical variables including spirometry. The SHQ alone was sensitive to differences in HRQL based on the number of involved organ systems. In conclusion, we have developed a self completed HRQL questionnaire for U.S. patients with sarcoidosis. PMID- 12738607 TI - Diagnosing pneumonia during mechanical ventilation: the clinical pulmonary infection score revisited. AB - The clinical pulmonary infection score-original or modified-has been proposed for the diagnosis and management of ventilator-associated pneumonia. In 79 episodes of suspected pneumonia, we prospectively assessed the diagnostic accuracy of the physicians' clinical assessment of probability and of the modified clinical pulmonary infection score, both measured before (pretest) and after (post-test) incorporating gram stains results, using bronchoalveolar lavage fluid culture as the reference test. The pretest clinical estimate was inaccurate (sensitivity 50%, specificity 58%); the mean clinical pulmonary infection score at baseline was 6.5 +/- 1.3 (range, 3-9) and 5.9 +/- 1.7 (range, 3-9), respectively, for the 40 confirmed and the 39 nonconfirmed episodes (p = 0.07), and only slightly more accurate (sensitivity 60%, specificity 59%) than the clinical prediction. Incorporating the gram stain results of either directed or blind protected sampling increased the diagnostic accuracy (sensitivity and specificity of 85% and 49% and 78% and 56%, respectively) of the clinical score and increased the likelihood ratio for pneumonia of a score of more than six from 1.46 to 1.67 and 1.77. The clinical pulmonary infection score has low diagnostic accuracy; however, incorporating gram stains results into the score may help clinical decision making in patients with clinically suspected pneumonia. PMID- 12738608 TI - Association of a missense mutation in the NOS3 gene with exhaled nitric oxide levels. AB - There is evidence that genetic factors affect nitric oxide formation and that sequence variants in the nitric oxide synthase genes contribute to the observed variance of nitric oxide levels in exhaled air (fraction of expired nitric oxide, FENO) in subjects with asthma. We identified a strong association between a known functional NOS3 missense sequence variant in the endothelial nitric oxide gene (G894T) and FENO level in a cohort of subjects with asthma. Age- and sex-adjusted FENO levels were lowest in asthmatic subjects with the TT genotype (geometric mean FENO [95% CI] = 7.17 [4.48 to 11.48] ppb) and were significantly higher in those with either the GT genotype (geometric mean FENO [95% CI] = 17.11 [13.80 to 21.23] ppb) or the GG genotype (geometric mean FENO [95% CI] = 12.06 [9.91 to 14.67] ppb) (F2,59 = 5.97, p = 0.004). The G894T DNA variant explained 16.3% of the residual variance in FENO levels. Our results demonstrate that the endothelial nitric oxide synthase, a nitric oxide synthase constitutively expressed in epithelial cells, plays an important role in determining measured levels of exhaled nitric oxide, a marker of the asthmatic condition. PMID- 12738609 TI - Glucose regulates lipid metabolism in fasting king penguins. AB - This study aims to determine whether glucose intervenes in the regulation of lipid metabolism in long-term fasting birds, using the king penguin as an animal model. Changes in the plasma concentration of various metabolites and hormones, and in lipolytic fluxes as determined by continuous infusion of [2-3H]glycerol and [1-14C]palmitate, were examined in vivo before, during, and after a 2-h glucose infusion under field conditions. All the birds were in the phase II fasting status (large fat stores, protein sparing) but differed by their metabolic and hormonal statuses, being either nonstressed (NSB; n = 5) or stressed (SB; n = 5). In both groups, glucose infusion at 5 mg.kg-1.min-1 induced a twofold increase in glycemia. In NSB, glucose had no effect on lipolysis (maintenance of plasma concentrations and rates of appearance of glycerol and nonesterified fatty acids) and no effect on the plasma concentrations of triacylglycerols (TAG), glucagon, insulin, or corticosterone. However, it limited fatty acid (FA) oxidation, as indicated by a 25% decrease in the plasma level of beta-hydroxybutyrate (beta-OHB). In SB, glucose infusion induced an approximately 2.5-fold decrease in lipolytic fluxes and a large decrease in FA oxidation, as reflected by a 64% decrease in the plasma concentration of beta-OHB. There were also a 35% decrease in plasma TAG, a 6.5- and 2.8-fold decrease in plasma glucagon and corticosterone, respectively, and a threefold increase in insulinemia. These data show that in fasting king penguins, glucose regulates lipid metabolism (inhibition of lipolysis and/or of FA oxidation) and affects hormonal status differently in stressed vs. nonstressed individuals. The results also suggest that in birds, as in humans, the availability of glucose, not of FA, is an important determinant of the substrate mix (glucose vs. FA) that is oxidized for energy production. PMID- 12738611 TI - Effects of peripheral CCK receptor blockade on food intake in rats. AB - Type A cholecystokinin receptor (CCKAR) antagonists differing in blood-brain barrier permeability were used to test the hypothesis that satiety is mediated, in part, by CCK action at CCKARs located peripheral to the blood-brain barrier. At dark onset, non-food-deprived rats received a bolus injection of devazepide (2.5 micromol/kg iv), a 3-h infusion of A-70104 (1 or 3 micromol x kg-1 x h-1 iv), or vehicle either alone or coadministered with a 3-h infusion of CCK-8 (10 nmol x kg-1 x h-1 iv) or a 2-h intragastric infusion of peptone (1 g/h). Food intake was determined from continuous computer recordings of changes in food bowl weight. Devazepide penetrates the blood-brain barrier; A-70104, the dicyclohexylammonium salt of Nalpha-3-quinolinoyl-d-Glu-N,N-dipentylamide (A 65186), does not. CCK-8 inhibited 3-h food intake by more than 50% and both A 70104 and devazepide abolished this response. A-70104 and devazepide stimulated food intake and similarly attenuated the anorexic response to intragastric infusion of peptone. Thus endogenous CCK appears to act, in part, at CCKARs peripheral to the blood-brain barrier to inhibit food intake. PMID- 12738610 TI - Drinking behavior elicited by central injection of angiotensin II: roles for protein kinase C and Ca2+/calmodulin-dependent protein kinase II. AB - Prior studies utilizing neurons cultured from the hypothalamus and brain stem of newborn rats have demonstrated that ANG II-induced modulation of neuronal firing involves activation of both protein kinase C (PKC) and Ca2+/calmodulin-dependent protein kinase II (CaMKII). The present studies were performed to determine whether these signaling molecules are also involved in physiological responses elicited by ANG II in the brain in vivo. Central injection of ANG II (10 ng/2 microl) into the lateral cerebroventricle (icv) of Sprague-Dawley rats increased water intake in a time-dependent manner. This ANG II-mediated dipsogenic response was attenuated by central injection of the PKC inhibitors chelerythrine chloride (0.5-50 microM, 2 microl) and Go-6976 (2.3 nM, 2 microl) and by the CaMKII inhibitor KN-93 (10 microM, 2 microl). Conversely, icv injection of chelerythrine chloride (50 microM, 2 microl) and KN-93 (10 microM, 2 microl) had no effect on the dipsogenic response elicited by central injection of carbachol (200 ng/2 microl). Furthermore, injection of ANG II (10 ng/2 microl) icv increases the activity of both PKC-alpha and CaMKII in rat septum and hypothalamus. These data suggest that signaling molecules involved in ANG II-induced responses in vitro are also relevant in physiological responses elicited by ANG II in the whole animal model. PMID- 12738612 TI - Physiological control of pituitary hormone secretory-burst mass, frequency, and waveform: a statistical formulation and analysis. AB - The present study investigates the time-varying control of pituitary hormone secretion over the day and night (D/N). To this end, we implemented an analytical platform designed to reconstruct simultaneously 1) basal (nonpulsatile) secretion, 2) single or dual secretory-burst waveforms, 3) random effects on burst amplitude, 4) stochastic pulse-renewal properties, 5) biexponential elimination kinetics, and 6) experimental uncertainty. The statistical solution is conditioned on a priori pulse-onset times, which are estimated in the first stage. Primary data composed of thyrotropin (TSH) concentrations were monitored over 24 h in 27 healthy adults. According to statistical criteria, 21/27 profiles favored a dual compared with single secretory-burst waveform. An objectively defined waveform change point (D/N boundary) emerged at 2046 (+/-23 min), after which 1) the mass of TSH released per burst increases by 2.1-fold (P < 0.001), 2) TSH secretory-burst frequency rises by 1.2-fold (P < 0.001), 3) the latency to maximal TSH secretion within a burst decreases by 67% (P < 0.001), 4) variability in secretory-burst shape diminishes by 50% (P < 0.001), and 5) basal TSH secretion declines by 17% (P < 0.002). In contrast, the regularity of successive burst times and the slow-phase half-life are stable. In conclusion, nycthemeral mechanisms govern TSH secretory-burst mass, frequency, waveform, and variability but not evidently TSH elimination kinetics or the pulse-timing process. Further studies will be required to assess the generality of the foregoing distinctive control mechanisms in other hypothalamo-pituitary axes. PMID- 12738613 TI - Single-fiber myosin heavy chain polymorphism: how many patterns and what proportions? AB - Previous studies have reported the existence of skeletal muscle fibers that coexpress multiple myosin heavy chain isoforms. These surveys have usually been limited to studying the polymorphic profiles of skeletal muscle fibers from a limited number of muscles (i.e., usually <4). Additionally, few studies have considered the functional implications of polymorphism. Hence, the primary objective of this study was to survey a relatively large number of rat skeletal muscle/muscle regions and muscle fibers (n approximately 5,000) to test the hypothesis that polymorphic fibers represent a larger fraction of the total pool of fibers than do so-called monomorphic fibers, which express only one myosin heavy chain isoform. Additionally, we used Hill's statistical model of the force velocity relationship to differentiate the functional consequences of single fiber myosin heavy chain isoform distributions found in these muscles. The results demonstrate that most muscles and regions of rodent skeletal muscles contain large proportions of polymorphic fibers, with the exception of muscles such as the slow soleus muscle and white regions of fast muscles. Several muscles were also found to have polymorphic profiles that are not consistent with the I<- >IIA<-->IIX<-->IIB scheme of muscle plasticity. For instance, it was found that the diaphragm muscle normally contains I/IIX fibers. Functionally, the high degree of polymorphism may 1) represent a strategy for producing a spectrum of contractile properties that far exceeds that simply defined by the presence of four myosin heavy chain isoforms and 2) result in relatively small differences in function as defined by the force-velocity relationship. PMID- 12738615 TI - Neuronal control of heart rate in isolated mouse atria. AB - A novel mouse isolated atrial preparation with intact postganglionic autonomic innervation was used to investigate the neuronal control of heart rate. To establish whether autonomic activation was likely to alter heart rate by modulating the hyperpolarization-activated current (If), the L-type Ca2+ current (ICa,L), or the ACh-activated K+ current (IK,ACh), the effects of nerve stimulation (right stellate ganglion or right vagus, 1-30 Hz) and autonomic agonists (0.1 microM norepinephrine or 0.3 microM carbachol) on heart rate were investigated in the presence of inhibitors of these currents, cesium chloride (Cs+, 1 mM), nifedipine (200 nM), and barium chloride (Ba2+, 0.1 mM), respectively. The positive chronotropic response to stellate ganglion stimulation was reduced by approximately 20% with Cs+ and nifedipine (P < 0.05), whereas the heart rate response to norepinephrine was only reduced with Cs+ (P < 0.05). Ba2+ attenuated the decrease in heart rate with vagal stimulation and carbachol by approximately 60% (P < 0.05). These results are consistent with the idea that sympathetic nerve stimulation modulates If to increase heart rate in the mouse. Activation of ICa,L also appears to contribute to the sympathetic heart rate response. However, the decrease in heart rate with vagal stimulation or carbachol is likely to result primarily from the activation of IK,ACh. PMID- 12738614 TI - Cardiac myofibroblasts isolated from the site of myocardial infarction express endothelin de novo. AB - Recently it was demonstrated that treatment with a nonselective endothelin (ET) receptor antagonist significantly reduces myocardial infarct size, which suggests a major role for ET in tissue repair following myocardial infarction (MI). Tissue repair and remodeling found at the site of MI are mainly attributed to myofibroblasts (myoFbs), which are phenotypically transformed fibroblasts that express alpha-smooth muscle actin. It is unclear whether myoFbs generate ET peptides and consequentially regulate pathophysiological functions de novo through expression of the ET-1 precursor (prepro-ET-1), ET-converting enzyme-1 (ECE-1), a metalloprotease that is required to convert Big ET-1 to ET-1 and ET receptors. To address these intriguing questions, we used cultured myoFbs isolated from 4-wk-old MI scar tissue. In cultured cells, we found: 1) expression of mRNA for ET precursor gene (ppET1), ECE-1, and ETA and ETB receptors by semiquantitative RT-PCR; 2) phosphoramidon-sensitive ECE-1 activity, which converts Big ET-1 to biologically active peptide ET-1; 3) expression of ETA and ETB receptors; 4) elaboration of Big ET-1 and ET-1 peptides in myoFb culture media; and 5) upregulation of type I collagen gene expression and synthesis by ET, which was blocked by bosentan (a nonselective ETA- and ETB receptor blocker). These studies clearly indicated that myoFbs express and generate ET-1 and receptor-mediated modulation of type I collagen expression by ET-1. Locally generated ET-1 may contribute to tissue repair of the infarcted heart in an autocrine/paracrine manner. PMID- 12738616 TI - Cardiovascular responses to substance P in the nucleus tractus solitarii: microinjection study in conscious rats. AB - The cardiovascular effects of substance P (SP) microinjections in the nucleus tractus solitarii (NTS) were evaluated in conscious rats. We chose this model because it is an effective way to access some of the cardiovascular effects of neurotransmitters in the NTS without the inconvenience of blunting pathways with anesthetic agents or removing forebrain projections by decerebration. The cardiovascular responses to SP injections were also evaluated after chronic nodose ganglionectomy. We found that, in conscious rats, SP microinjections into the NTS induced hypertension and tachycardia. Unilateral and bilateral SP injections into the NTS caused a slow increase in blood pressure and heart rate that peaked 1.5-5 min after injection and lasted for 20-30 min. Nodose ganglionectomy increased the duration of the pressor and tachycardic effects of SP and enhanced the pressor response. These data show that SP in the NTS is involved in pressor pathways. The supersensitivity to SP seen after nodose ganglionectomy suggests that vagal afferent projections are involved in those pressor pathways activated by SP in the NTS. PMID- 12738618 TI - Low arterial compliance in young African-American males. AB - Hypertension remains a common public health challenge because of its prevalence and increase in co-morbid cardiovascular diseases. Black males have disproportionate pathophysiological consequences of hypertension compared with any other group in the United States. Alterations in arterial wall compliance and autonomic function often precede the onset of disease. Accordingly, our purpose was to investigate whether differences exist in arterial compliance and autonomic function between young, healthy African-American males without evidence of hypertension and age- and gender-matched non-African-American males. All procedures were carried out noninvasively following rest. Arterial compliance was calculated as the integrated area starting at the well-defined nadir of the incisura of the dicrotic notch to the end of diastole of the radial artery pulse wave. Power spectral analysis of heart rate and blood pressure variability provided distributions representative of parasympathetic and sympathetic modulations and sympathovagal balance. Baroreflex sensitivity (BRS) was calculated using the sequence method. Thirty-two African-American and twenty-nine non-African-American males were comparable in anthropometrics and negative family history of hypertension. t-Tests revealed lower arterial compliance (5.8 +/- 2.4 vs. 8.6 +/- 4.0 mmHg. s; P = 0.0017), parasympathetic modulation (8.9 +/- 1.1 vs. 9.7 +/- 1.1 ln ms2; P = 0.0063), and BRS (13.7 +/- 7.3 vs. 21.1 +/- 8.5 ms/mmHg; P = 0.0007) and higher sympathovagal balance (2.9 +/- 3.2 vs. 1.5 +/- 1.1; P = 0.03) in the African-American group. In summary, differences exist in arterial compliance and autonomic balance in African-American males. These alterations may be antecedent markers of disease and valuable in the detection of degenerative cardiovascular processes in individuals at risk. PMID- 12738617 TI - Transient transcapillary exchange of water driven by osmotic forces in the heart. AB - Osmotic transient responses in organ weight after changes in perfusate osmolarity have implied steric hindrance to small-molecule transcapillary exchange, but tracer methods do not. We obtained osmotic weight transient data in isolated, Ringer-perfused rabbit hearts with NaCl, urea, glucose, sucrose, raffinose, inulin, and albumin and analyzed the data with a new anatomically and physicochemically based model accounting for 1) transendothelial water flux, 2) two sizes of porous passages across the capillary wall, 3) axial intracapillary concentration gradients, and 4) water fluxes between myocytes and interstitium. During steady-state conditions approximately 28% of the transcapillary water flux going to form lymph was through the endothelial cell membranes [capillary hydraulic conductivity (Lp) = 1.8 +/- 0.6 x 10-8 cm. s-1. mmHg-1], presumably mainly through aquaporin channels. The interendothelial clefts (with Lp = 4.4 +/- 1.3 x 10-8 cm. s-1. mmHg-1) account for 67% of the water flux; clefts are so wide (equivalent pore radius was 7 +/- 0.2 nm, covering approximately 0.02% of the capillary surface area) that there is no apparent hindrance for molecules as large as raffinose. Infrequent large pores account for the remaining 5% of the flux. During osmotic transients due to 30 mM increases in concentrations of small solutes, the transendothelial water flux was in the opposite direction and almost 800 times as large and was entirely transendothelial because no solute gradient forms across the pores. During albumin transients, gradients persisted for long times because albumin does not permeate small pores; the water fluxes per milliosmolar osmolarity change were 200 times larger than steady-state water flux. The analysis completely reconciles data from osmotic transient, tracer dilution, and lymph sampling techniques. PMID- 12738619 TI - Pattern formation of vascular smooth muscle cells subject to nonuniform fluid shear stress: role of PDGF-beta receptor and Src. AB - Blood vessels are subject to fluid shear stress, a hemodynamic factor that inhibits the mitogenic activities of vascular cells. The presence of nonuniform shear stress has been shown to exert graded suppression of cell proliferation and induces the formation of cell density gradients, which in turn regulate the direction of smooth muscle cell (SMC) migration and alignment. Here, we investigated the role of platelet-derived growth factor (PDGF)-beta receptor and Src in the regulation of such processes. In experimental models with vascular polymer implants, SMCs migrated from the vessel media into the neointima of the implant under defined fluid shear stress. In a nonuniform shear model, blood shear stress suppressed the expression of PDGF-beta receptor and the phosphorylation of Src in a shear level-dependent manner, resulting in the formation of mitogen gradients, which were consistent with the gradient of cell density as well as the alignment of SMCs. In contrast, uniform shear stress in a control model elicited an even influence on the activity of mitogenic molecules without modulating the uniformity of cell density and did not significantly influence the direction of SMC alignment. The suppression of the PDGF-beta receptor tyrosine kinase and Src with pharmacological substances diminished the gradients of mitogens and cell density and reduced the influence of nonuniform shear stress on SMC alignment. These observations suggest that PDGF-beta receptor and Src possibly serve as mediating factors in nonuniform shear-induced formation of cell density gradients and alignment of SMCs in the neointima of vascular polymer implants. PMID- 12738620 TI - Improved oxygenation in ischemic hamster flap tissue is correlated with increasing hemodilution with Hb vesicles and their O2 affinity. AB - The aim of this study was to test the influence of oxygen affinity of Hb vesicles (HbVs) and level of blood exchange on the oxygenation in collateralized, ischemic, and hypoxic hamster flap tissue during normovolemic hemodilution. Microhemodynamics were investigated with intravital microscopy. Tissue Po2 was measured with Clark-type microprobes. HbVs with a P50 of 15 mmHg (HbV15) and 30 mmHg (HbV30) were suspended in 6% Dextran 70 (Dx70). The Hb concentration of the solutions was 7.5 g/dl. A stepwise replacement of 15%, 30%, and 50% of total blood volume was performed, which resulted in a gradual decrease in total Hb concentration. In the ischemic tissue, hemodilution led to an increase in microvascular blood flow to maximally 141-166% of baseline in all groups (median; P < 0.01 vs. baseline, not significant between groups). Oxygen tension was transiently raised to 121 +/- 17% after the 30% blood exchange with Dx70 (P < 0.05), whereas it was increased after each step of hemodilution with HbV15-Dx70 and HbV30-Dx70, reaching 217 +/- 67% (P < 0.01) and 164 +/- 33% (P < 0.01 vs. baseline and other groups), respectively, after the 50% blood exchange. We conclude that despite a decrease in total Hb concentration, the oxygenation in the ischemic, hypoxic tissue could be improved with increasing blood exchange with HbV solutions. Furthermore, better oxygenation was obtained with the left shifted HbVs. PMID- 12738621 TI - Oxygen delivery to skeletal muscle fibers: effects of microvascular unit structure and control mechanisms. AB - The number of perfused capillaries in skeletal muscle varies with muscle activation. With increasing activation, muscle fibers are recruited as motor units consisting of widely dispersed fibers, whereas capillaries are recruited as groups called microvascular units (MVUs) that supply several adjacent fibers. In this study, a theoretical model was used to examine the consequences of this spatial mismatch between the functional units of muscle activation and capillary perfusion. Diffusive oxygen transport was simulated in cross sections of skeletal muscle, including several MVUs and fibers from several motor units. Four alternative hypothetical mechanisms controlling capillary perfusion were considered. First, all capillaries adjacent to active fibers are perfused. Second, all MVUs containing capillaries adjacent to active fibers are perfused. Third, each MVU is perfused whenever oxygen levels at its feed arteriole fall below a threshold value. Fourth, each MVU is perfused whenever the average oxygen level at its capillaries falls below a threshold value. For each mechanism, the dependence of the fraction of perfused capillaries on the level of muscle activation was predicted. Comparison of the results led to the following conclusions. Control of perfusion by MVUs increases the fraction of perfused capillaries relative to control by individual capillaries. Control by arteriolar oxygen sensing leads to poor control of tissue oxygenation at high levels of muscle activation. Control of MVU perfusion by capillary oxygen sensing permits adequate tissue oxygenation over the full range of activation without resulting in perfusion of all MVUs containing capillaries adjacent to active fibers. PMID- 12738622 TI - Vascular and metabolic response to isolated small muscle mass exercise: effect of age. AB - To determine the effect of age on quadriceps muscle blood flow (QMBF), leg vascular resistance (LVR), and maximum oxygen uptake (QVO2 max), a thermal dilution technique was used in conjunction with arterial and venous femoral blood sampling in six sedentary young (19.8 +/- 1.3 yr) and six sedentary old (66.5 +/- 2.1 yr) males during incremental knee extensor exercise (KE). Young and old attained a similar maximal KE work rate (WRmax) (young: 25.2 +/- 2.1 and old: 24.1 +/- 4 W) and QVO2 max (young: 0.52 +/- 0.03 and old: 0.42 +/- 0.05 l/min). QMBF during KE was lower in old subjects by approximately 500 ml/min across all work rates, with old subjects demonstrating a significantly lower QMBF/W (old: 174 +/- 20 and young: 239 +/- 46 ml. min-1. W-1). Although the vasodilatory response to incremental KE was approximately 142% greater in the old (young: 0.0019 and old: 0.0046 mmHg. min. ml-1. W-1), consistently elevated leg vascular resistance (LVR) in the old, approximately 80% higher LVR in the old at 50% WR and approximately 40% higher LVR in the old at WRmax (young: 44.1 +/- 3.6 and old: 31.0 +/- 1.7 mmHg. min. ml-1), dictated that during incremental KE the LVR of the old subjects was never less than that of the young subjects. Pulse pressures, indicative of arterial vessel compliance, were approximately 36% higher in the old subjects across all work rates. In conclusion, well-matched sedentary young and old subjects with similar quadriceps muscle mass achieved a similar WRmax and QVO2 max during incremental KE. The old subjects, despite a reduced QMBF, had a greater vasodilatory response to incremental KE. Given that small muscle mass exercise, such as KE, utilizes only a fraction of maximal cardiac output, peripheral mechanisms such as consistently elevated leg vascular resistance and greater pulse pressures appear to be responsible for reduced blood flow persisting throughout graded KE in the old subjects. PMID- 12738623 TI - Minimally invasive aortic banding in mice: effects of altered cardiomyocyte insulin signaling during pressure overload. AB - We developed a minimally invasive method for producing left ventricular (LV) pressure overload in mice. With the use of this technique, we quickly and reproducibly banded the transverse aorta with low surgical morbidity and mortality. Minimally invasive transverse aortic banding (MTAB) acutely and chronically increased LV systolic pressure, increased heart weight-to-body weight ratio, and induced myocardial fibrosis. We used this technique to determine whether reduced insulin signaling in the heart altered the cardiac response to pressure overload. Mice with cardiac myocyte-restricted knockout of the insulin receptor (CIRKO) have smaller hearts than wild-type (WT) controls. Four weeks after MTAB, WT and CIRKO mice had comparably increased LV systolic pressure, increased cardiac mass, and induction of mRNA for beta-myosin heavy chain and atrial natriuretic factor. However, CIRKO hearts were more dilated, had depressed LV systolic function by echocardiography, and had greater interstitial fibrosis than WT mice. Expression of connective tissue growth factor was increased in banded CIRKO hearts compared with WT hearts. Thus lack of insulin signaling in the heart accelerates the transition to a more decompensated state during cardiac pressure overload. The use of the MTAB approach should facilitate the study of the pathophysiology and treatment of pressure-overload hypertrophy. PMID- 12738624 TI - Hemodynamic and renal effects of low-dose brain natriuretic peptide infusion in humans: a randomized, placebo-controlled crossover study. AB - Brain natriuretic peptide (BNP) is a cardiac hormone with natriuretic activity. The aim of this study was to investigate the cardiovascular effects of pathophysiological levels of BNP on central hemodynamics, cardiac function, renal hemodynamics and function, and microvascular hemodynamics in healthy subjects. In this double-blind, placebo-controlled crossover study, we intravenously infused BNP (4 pmol. kg-1. min-1) or placebo for 1 h on two separate days in 12 healthy subjects (mean age, 60 +/- 5 yr). Nailfold and conjunctival capillary density, finger-skin (thermoregulatory) microvascular blood flow, and cardiac output were studied before and after infusion using intravital videomicroscopy, laser-Doppler fluxmetry, and echocardiography, respectively. Furthermore, during infusion, we measured the effective renal plasma flow and glomerular filtration rate using p aminohippurate and inulin clearances. Blood pressure and heart rate were monitored for all measurements. Compared with placebo, BNP significantly decreased stroke volume with a tendency to decrease cardiac output. With subjects in the sitting position, mean arterial pressure decreased and heart rate increased after BNP infusion, whereas with subjects in the supine position, these variables remained unchanged. BNP increased natriuresis, diuresis, glomerular filtration rate, filtration fraction, and filtered load of Na+ compared with placebo, whereas effective renal plasma flow did not change. BNP did not affect the microvascular capillary density of conjunctiva and skin, microvascular blood flow, total skin oxygen capacity, and postocclusive recruitment. These results suggest that BNP has predominantly central and renal hemodynamic effects; however, it does not influence peripheral microcirculation in skin and conjunctiva. PMID- 12738625 TI - Molecular and pharmacological approaches to inhibiting nitric oxide after burn trauma. AB - Whereas controversial, several studies have suggested that nitric oxide (NO) alters cardiac contractility via cGMP, peroxynitrite, or poly(ADP ribose) synthetase (PARS) activation. This study determined whether burn-related upregulation of myocardial inducible NO synthase (iNOS) and NO generation contributes to burn-mediated cardiac contractile dysfunction. Mice homozygous null for the iNOS gene (iNOS knockouts) were obtained from Jackson Laboratory. iNOS knockouts (KO) as well as wild-type mice were given a cutaneous burn over 40% of the total body surface area by the application of brass probes (1 x 2 x 0.3 cm) heated to 100 degrees C to the animals' sides and back for 5 s (iNOS/KO burn and wild-type burn). Additional groups of iNOS KO and wild-type mice served as appropriate sham burn groups (iNOS/KO sham and wild-type sham). Cardiac function was assessed 24 h postburn by perfusing hearts (n = 7-10 mice/group). Burn trauma in wild-type mice impaired cardiac function as indicated by the lower left ventricular pressure (LVP, 67 +/- 2 mmHg) compared with that measured in wild-type shams (94 +/- 2 mmHg, P < 0.001), a lower rate of LVP rise (+dP/dtmax, 1,620 +/- 94 vs. 2,240 +/- 58 mmHg/s, P < 0.001), and a lower rate of LVP fall ( dP/dtmax, 1,200 +/- 84 vs. 1,800 +/- 42 mmHg/s, P < 0.001). Ventricular function curves confirmed significant contractile dysfunction after burn trauma in wild type mice. Burn trauma in iNOS KO mice produced fewer cardiac derangements compared with those observed in wild-type burns (LVP: 78 +/- 5 mmHg; +dP/dt: 1,889 +/- 160 mmHg/s; -dP/dt: 1,480 +/- 154 mmHg/s). The use of a pharmacological approach to inhibit iNOS (aminoguanidine, given ip) in additional wild-type shams and burns confirmed the iNOS KO data. Whereas the absence of iNOS attenuated burn mediated cardiac contractile dysfunction, these experiments did not determine the contribution of cardiac-derived NO versus NO generated by immune cells. However, our data indicate a role for NO in cardiac dysfunction after major trauma. PMID- 12738626 TI - Endothelium-dependent responses in coronary arteries are changed with puberty in male pigs. AB - In humans, cardiovascular disease begins in young adulthood and is more prevalent in males than females. However, little is known about vascular function during transition to adulthood in males. The aim of this study was to define changes in production of endothelium-derived nitric oxide (NO) and coronary arterial responses during puberty. Plasma was collected from juvenile (2-3 mo of age) and adult (5-6 mo of age) male pigs (n = 8/group) for measurement of NO, and aortic endothelial cells were collected for measurement of mRNA and protein for endothelial NO synthase (eNOS). Although plasma NO was higher in juvenile (67.0 +/- 25.6 microM) than in adult (15.0 +/- 7.1 microM) male pigs, eNOS protein was similar in both groups. However, levels of mRNA for eNOS were lower in aortic endothelial cells from juvenile pigs. In rings of coronary arteries suspended in organ chambers for measurement of isometric force and contracted with PGF2alpha, relaxations to an alpha2-adrenergic agonist were significantly inhibited by indomethacin only in juvenile pigs [EC50 (-log M), 6.7 +/- 0.3 with indomethacin and 7.7 +/- 0.3 under control conditions]. NG-monomethyl-l-arginine (l-NMMA) inhibited relaxations in both groups. On the contrary, in the presence of indomethacin, relaxations to bradykinin were inhibited by l-NMMA only in arteries from adult pigs [EC50 (-log M), 8.9 +/- 0.3 with indomethacin and 8.6 +/- 0.3 with addition of l-NMMA]. These results suggest that hormonal changes associated with sexual maturity may affect posttranscriptional and/or translational regulation of eNOS protein and result in lower plasma NO in adult male pigs. In addition, endothelium-derived inhibitory cyclooxygenase products seem to predominate in juveniles. PMID- 12738627 TI - Clinical immunology and traditional herbal medicines. PMID- 12738628 TI - Production of genetically engineered biotinylated interleukin-2 and its application in a rapid nonradioactive assay for T-cell activation. AB - The development of reliable assay systems that can measure lymphocyte activation in vitro has been a major goal of immunodiagnostics. Traditionally, tritiated thymidine incorporation has been used to monitor T-cell activation. Other methods include enzyme-linked immunosorbent assay (ELISA), enzyme-linked immunospot assay, and colorimetric assays. We have established a lymphocyte activation assay that utilizes fluorescein isothiocyanate (FITC)-streptavidin bound to recombinant biotinylated human interleukin-2 (IL-2). Utilizing recombinant DNA technology, a unique monobiotinylated human IL-2 has been created and isolated using the Promega PinPoint vector system. ELISA has been used to demonstrate streptavidin binding and recognition by a human IL-2-specific antibody. IL-2 function has been demonstrated using a murine IL-2-dependent T-cell line, CTLL-2, responsive to human IL-2. Recombinant biotinylated human IL-2 conjugated to streptavidin-FITC or streptavidin-horseradish peroxidase has been used to monitor T-cell activation in the presence of antigen as well as mitogen. The sensitivity and convenience of this method make this lymphocyte activation assay an attractive alternative to tritiated thymidine incorporation as a method for monitoring T-cell activation. In addition, the availability of a recombinant biotinylated human IL-2 will permit the production of a uniform product suitable for diagnostic and clinical application. PMID- 12738629 TI - Immunoglobulin G subisotype responses of pneumonic and healthy, exposed foals and adult horses to Rhodococcus equi virulence-associated proteins. AB - Rhodococcus equi causes severe pyogranulomatous pneumonia in foals and in immunocompromised humans. Replication of virulent isolates within macrophages correlates with the presence of a large plasmid which encodes a family of seven virulence-associated proteins (VapA and VapC to VapH), whose functions are unknown. Although cell-mediated immunity is thought to be crucial in eliminating R. equi infection, antibody partially protects foals. The antibody response to both VapA and VapC was similar in six adult horses and six naturally exposed but healthy foals, as well as in eight foals with R. equi pneumonia. The immunoglobulin G (IgG) subisotype response of pneumonic foals to Vap proteins was significantly IgGb biased and also had a trend toward higher IgGT association compared to the isotype association of antibody in adult horses and healthy exposed foals. This suggests that in horses, IgGb and IgGT are Th2 isotypes and IgGa is a Th1 isotype. Furthermore, it suggests that foals which develop R. equi pneumonia have a Th2-biased, ineffective immune response whereas foals which become immune develop a Th1-biased immune response. Pneumonic foals had significantly more antibody to VapD and VapE than did healthy exposed foals. This may indicate a difference in the expression of these two Vap proteins during persistent infection. Alternatively, in pneumonic foals the deviation of the immune response toward VapD and VapE may reflect a bias unfavorable to R. equi resistance. These data indicate possible age-related differences in the equine immune response affecting Th1-Th2 bias as well as antibody specificity bias, which together favor the susceptibility of foals to R. equi pneumonia. PMID- 12738630 TI - Respiratory disease in kennelled dogs: serological responses to Bordetella bronchiseptica lipopolysaccharide do not correlate with bacterial isolation or clinical respiratory symptoms. AB - The role of Bordetella bronchiseptica in a natural outbreak of canine infectious respiratory disease was investigated both by culture and serological analysis. B. bronchiseptica was found in the lungs of a large proportion of clinically healthy dogs and in a greater proportion of dogs with respiratory disease. Using a lipopolysaccharide (LPS) antigen-based enzyme-linked immunosorbent assay, we analyzed the serological responses of a large number of dogs. Dogs with high antibody levels showed no protection from disease, and there was no correlation between the development of disease and rising antibody titer. Similarly, there was no difference in antibody levels in dogs with and without B. bronchiseptica in the lungs. Antibodies to LPS have no predictive value in determining which animals will contract respiratory disease, how severe the disease will be, or which dogs will have B. bronchiseptica colonizing the lungs. PMID- 12738631 TI - Effects of sample collection and storage methods on antipneumococcal immunoglobulin A in saliva. AB - Saliva contains components of both the mucosal and systemic immune systems. Variable flow rates, immunoglobulin proteases, and variation in collection and storage methods all introduce differences in the estimated concentrations of antibodies. We evaluated the effect of four collection methods and three storage protocols on the concentrations of immunoglobulin A (IgA) antibodies to pneumococcal capsular antigens 1, 5, 6B, and 14 and to pneumococcal surface adhesin A (PsaA) in saliva. Specimens were collected from 30 healthy Kenyan adults by collecting drool, by pipette suction, and with two commercial kits, OraSure and Oracol. Aliquots from each specimen were snap-frozen with glycerol in liquid nitrogen or stored for 4 to 8 h at +4 degrees C either with or without the addition of protease enzyme inhibitors prior to storage at -70 degrees C. Anticapsular IgA concentrations were not significantly different with different collection methods, but snap-freezing the specimens in liquid nitrogen led to concentrations 41 to 47% higher than those of specimens stored by the other methods (P < 0.0005). PMID- 12738632 TI - Relationship between plasma Interleukin-12 (IL-12) and IL-18 levels and severe malarial anemia in an area of holoendemicity in western Kenya. AB - In this study, we investigated whether levels of interleukin-12 (IL-12) and IL-18 in plasma are associated with severe malarial anemia outcomes in an area of holoendemicity in western Kenya. We compared plasma IL-12 and IL-18 levels in six groups of children grouped into the categories aparasitemic, asymptomatic, mild malaria, high-density uncomplicated malaria (UC), moderate malarial anemia (MMA), or severe malarial anemia (SMA). IL-12 levels were significantly reduced in children with SMA (P < 0.05) but not in other groups compared to children in the aparasitemic control group. IL-18, a cytokine known to be critical for the induction of gamma interferon along with IL-12, was produced more frequently (70%) in children with UC (P = 0.06) than in children in the aparasitemic control group (32%). However, in the SMA group the IL-18 response rate declined to 30%, which was similar to that in the aparasitemic control group, which showed a 32% response rate. This finding suggests that the IL-18 response may be impaired in children with SMA. In summary, the results from this study support the hypothesis that impairment of IL-12 and/or IL-18 response may contribute to the development of severe malarial anemia in areas of holoendemicity for malaria. PMID- 12738633 TI - Production of Chlamydia pneumoniae proteins in Bacillus subtilis and their use in characterizing immune responses in the experimental infection model. AB - Due to intracellular growth requirements, large-scale cultures of chlamydiae and purification of its proteins are difficult and laborious. To overcome these problems we produced chlamydial proteins in a heterologous host, Bacillus subtilis, a gram-positive nonpathogenic bacterium. The genes of Chlamydia pneumoniae major outer membrane protein (MOMP), the cysteine-rich outer membrane protein (Omp2), and the heat shock protein (Hsp60) were amplified by PCR, and the PCR products were cloned into expression vectors containing a promoter, a ribosome binding site, and a truncated signal sequence of the alpha-amylase gene from Bacillus amyloliquefaciens. C. pneumoniae genes were readily expressed in B. subtilis under the control of the alpha-amylase promoter. The recombinant proteins MOMP and Hsp60 were purified from the bacterial lysate with the aid of the carboxy-terminal histidine hexamer tag by affinity chromatography. The Omp2 was separated as an insoluble fraction after 8 M urea treatment. The purified proteins were successfully used as immunogens and as antigens in serological assays and in a lymphoproliferation test. The Omp2 and Hsp60 antigens were readily recognized by the antibodies appearing after pulmonary infection following intranasal inoculation of C. pneumoniae in mice. Also, splenocytes collected from mice immunized with MOMP or Hsp60 proteins proliferated in response to in vitro stimulation with the corresponding proteins. PMID- 12738634 TI - NF-kappaB is involved in regulation of CD40 ligand expression on Mycobacterium bovis bacillus Calmette-Guerin-activated human T cells. AB - Interaction between CD40L (CD154) on activated T cells and its receptor CD40 on antigen-presenting cells has been reported to be important in the resolution of infection by mycobacteria. However, the mechanism(s) by which Mycobacterium bovis bacillus Calmette-Guerin (BCG) up-regulates membrane expression of CD40L molecules is poorly understood. This study was done to investigate the role of the nuclear factor kappaB (NF-kappaB) signaling pathway in the regulation of CD40L expression in human CD4(+) T cells stimulated with BCG. Specific pharmacologic inhibition of the NF-kappaB pathway revealed that this signaling cascade was required in the regulation of CD40L expression on the surface of BCG activated CD4(+) T cells. These results were further supported by the fact that treatment of BCG-activated CD4(+) T cells with these pharmacological inhibitors significantly down-regulated CD40L mRNA. In this study, inhibitor kappaBalpha (IkappaBalpha) and IkappaBbeta protein production was not affected by the chemical protease inhibitors and, more importantly, BCG led to the rapid but transient induction of NF-kappaB activity. Our results also indicated that CD40L expression on BCG-activated CD4(+) T cells resulted from transcriptional up regulation of the CD40L gene by a mechanism which is independent of de novo protein synthesis. Interestingly, BCG-induced activation of NF-kappaB and the increased CD40L cell surface expression were blocked by the protein kinase C (PKC) inhibitors 1-[5-isoquinolinesulfonyl]-2-methylpiperazine and salicylate, both of which block phosphorylation of IkappaB. Moreover, rottlerin a Ca(2+) independent PKC isoform inhibitor, significantly down-regulated CD40L mRNA in BCG activated CD4(+) T cells. These data strongly suggest that CD40L expression by BCG-activated CD4(+) T cells is regulated via the PKC pathway and by NF-kappaB DNA binding activity. PMID- 12738635 TI - Humoral immune responses to S-layer-like proteins of Bacteroides forsythus. AB - Bacteroides forsythus is one of the important periodontopathic bacteria, and this microorganism is known to have an S-layer outside the outer membrane. The S-layer like antigens were recently isolated from B. forsythus, and they were found to be 270- and 230-kDa proteins in the envelope fraction. In this study, these proteins were confirmed to be specific to B. forsythus by sodium dodecyl sulfate polyacrylamide gel electrophoresis, and they were clearly recognized by sera from patients with adult and early-onset periodontitis in Western immmunoblot analysis. We compared the immunoglobulin G (IgG) responses against the purified S layer-like antigen by enzyme-linked immunosorbent assay. IgG responses against this antigen were low in healthy control subjects, but they were significantly higher in subjects with adult and early-onset periodontitis. Together with the fact that the IgG responses against the crude extract of B. forsythus did not rise significantly in patients with periodontitis, S-layer-like proteins are considered to be specific antigens of B. forsythus and may play an important role in the progression of periodontitis. PMID- 12738636 TI - Cervicovaginal neutralizing antibodies to herpes simplex virus (HSV) in women seropositive for HSV Types 1 and 2. AB - Antibodies to herpes simplex virus type 1 (HSV-1) and HSV-2 of the immunoglobulin G (IgG) and IgA isotypes were detected in the cervicovaginal secretions (CVS) of 77 HSV-1- and HSV-2-seropositive but clinically asymptomatic African women by type-specific enhanced chemiluminescence Western blotting (ECL-WB). Of the 77 subjects, 34 were HIV negative, shedding HSV-2 DNA in their genital secretions; 20 were HIV positive, shedding HSV-2 DNA; and 23 were HIV negative, not shedding HSV-2 DNA. HSV-specific IgG was detected in CVS of nearly 70% of the women studied. HSV-specific IgA was found in CVS of 50% of the women studied. The distribution of CVS HSV-specific antibodies to each HSV type was highly heterogeneous, with a slight predominance of detectable IgG to HSV-1 (59%) over IgG to HSV-2 (41%), whereas the frequency of detectable IgA to HSV-1 (39%) was similar to that of IgA to HSV-2 (36%). The presence of detectable HSV-specific antibodies was inversely associated with HSV-2 DNA genital asymptomatic shedding but was not affected by HIV seropositivity. In addition, 13 of 77 (17%) CVS samples showed neutralizing activity against HSV-2, as assessed by an HSV-2 in vitro infectivity reduction assay. Neutralizing activity in CVS was associated with the presence of IgG and/or IgA antibodies to HSV-1 and/or to HSV-2 by ECL WB. Among women whose CVS showed HSV-2-neutralizing activity, the specific activity of HSV-specific neutralizing antibodies was substantially (fivefold) higher in HSV-2 DNA shedders than in nonshedders. In conclusion, HSV-specific antibodies are frequently detected in CVS of asymptomatic African women seropositive for HSV-1 and HSV-2. A subset of these women had functional neutralizing activity against HSV-2 in their CVS. The origin of these antibodies and their role in HSV-2 disease of the female genital tract remain to be determined. PMID- 12738637 TI - Evaluation of an immunoglobulin M capture enzyme-linked immunosorbent assay for diagnosis of Orientia tsutsugamushi infection. AB - To differentiate scrub typhus from other acute febrile diseases, a rapid and reliable serological diagnosis is important. We developed an immunoglobulin M (IgM) capture enzyme-linked immunosorbent assay (ELISA) for diagnosis of recent Orientia tsutsugamushi infections in humans. The 56-kDa major outer membrane protein of O. tsutsugamushi is well known as the most immunodominant antigen in scrub typhus. The test is based on the use of the biotinylated recombinant 56-kDa protein of O. tsutsugamushi Boryong, Bor56, which was expressed as a fusion protein with a maltose-binding protein in Escherichia coli. In the test, the serum IgM antibodies were captured by anti-human IgM antibodies coated onto a microtiter plate. The captured IgM antibodies were revealed through sequential addition of biotinylated Bor56 antigen and peroxidase-conjugated streptavidin to the plate. The IgM capture ELISA was compared with the immunofluorescence antibody assay (IFA) by testing 176 serum samples from patients with diagnosed cases of rickettsial disease and patients with other acute febrile diseases. Of the 81 IgG IFA-positive samples, 78 tested positive (sensitivity, 96.3%) and all 31 IgM IFA-positive samples tested positive (sensitivity, 100%) by the IgM capture ELISA. The specificity of the IgM capture ELISA was 99%, and 1 of the 95 IFA-negative samples was positive in the assay. These results strongly suggest that IgM capture ELISA using the recombinant Bor56 antigen is a reliable and detailed method for the detection of early O. tsutsugamushi infection. PMID- 12738638 TI - TH1 and TH2 cytokine mRNA and protein levels in human immunodeficiency virus (HIV)-seropositive and HIV-seronegative youths. AB - The roles of cytokines in the progression of human immunodeficiency virus (HIV) associated disease are controversial. The patterns of innate cytokine production have been postulated to shift from TH1- to TH2-type cytokines with the progression of HIV-associated disease. Although there have been studies of cytokines in children and adults, no data are available on cytokine production in healthy or HIV-infected adolescents. We analyzed and characterized cytokine mRNA and protein levels for gamma interferon, interleukin 2 (IL-2), IL-4, and tumor necrosis factor alpha and protein levels of IL-6 in both stimulated and unstimulated peripheral blood mononuclear cells obtained from a large longitudinal, observational cohort study of HIV-seropositive and -seronegative adolescents. We correlated cytokine results with viral load and CD4(+)-T-cell counts as critical markers of disease progression in HIV-infected adolescents. These data were used to examine hypotheses related to the TH1-to-TH2 cytokine shift in a sample of HIV-infected adolescents. Five hundred twenty subjects participating in the REACH (Reaching for Excellence in Adolescent Care and Health) Project of the Adolescent Medicine HIV/AIDS Research Network contributed blood samples. Samples selected for the cross-sectional data set analyzed had to meet selection criteria developed to minimize the potential confounding effects of acute intercurrent illnesses or infections, recent vaccination for hepatitis, and altered hormone status and to optimize congruence of cytokine measurements with assays of viral load and CD4(+)-T-cell counts. Group differences in the proportions of subjects with detectable levels of each cytokine marker were compared. In the subset of subjects with detectable cytokine values, differences in detected values were compared across subgroups defined by HIV serostatus and among HIV-seropositive subjects by three viral load classifications. The study sample was 65% HIV seropositive, 71% African-American, and 75% female with a mean age of 17.4 years. HIV-seropositive subjects were relatively healthy with mean and median CD4(+)-T-cell counts of 534 and 499 cells/mm(3), respectively. Only 8.1% of subjects had CD4(+)-T-cell counts below 200 cells/mm(3), and 25% had viral loads that were below the threshold of detection (<400 copies/ml). Detailed analyses of these data indicate that there were no differences in cytokines detected in HIV-seropositive and HIV-seronegative adolescents, and there was no apparent relationship between the cytokine measurements and the viral load or CD4(+)-T-cell categorization, the parameters selected as markers of HIV associated disease status. These adolescents, including the HIV-seropositive subjects, were relatively healthy, and the HIV-infected subjects were at an early stage in the course of their HIV-associated disease. On the basis of our data, we conclude that, early in the course of HIV-associated disease in adolescents, there are no detectable shifts from TH1 to TH2 cytokine production. PMID- 12738639 TI - MD-2 is necessary for the toll-like receptor 4 protein to undergo glycosylation essential for its translocation to the cell surface. AB - MD-2 has been reported to be required for the translocation of the Toll-like receptor 4 (TLR4) to the cell surface. However, the mechanism by which MD-2 promotes TLR4 translocation is unknown. We identified the presence of two forms of TLR4 with different molecular masses (approximately 110 and 130 kDa) when TLR4 was expressed together with MD-2. Expressing TLR4 alone produced only the 110-kDa form. Using a membrane-impermeable biotinylation reagent, we found that only the 130-kDa form of TLR4 was expressed on the cell surface. When a cellular extract prepared from cells expressing TLR4 and MD-2 was treated with N-glycosidase, the two forms of TLR4 converged into a single band whose size was smaller than the 110-kDa form of TLR4. Mutation of TLR4 at Asn(526) or Asn(575) resulted in the disappearance of the 130-kDa form and prevented TLR4 from being expressed on the cell surface without affecting the ability of TLR4 to associate with MD-2. These results indicate that TLR4 is able to undergo multiple glycosylations without MD 2 but that the specific glycosylation essential for cell surface expression requires the presence of MD-2. PMID- 12738641 TI - Role of macrophage migration inhibitory factor in otitis media with effusion in adults. AB - Otitis media with effusion (OME) is one of the most common ear diseases. Bacterial endotoxins and several inflammatory cytokines appear to be involved in the pathogenesis of OME in children; however, little is known of the immunological aspects of the onset of OME in adults. We sought to determine the presence of macrophage migration inhibitory factor (MIF) as well as interleukin 1beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha), RANTES (regulated upon activation, normal T-cell expressed and presumably secreted), and endotoxin in middle ear effusions (MEEs) from adult patients with OME. In addition, the levels of MIF in MEEs from adults and children were compared. MEE was obtained from 95 adults and 11 children. The levels of MIF, IL-1beta, TNF-alpha, and RANTES were determined by enzyme-linked immunosorbent assay, and the concentrations of endotoxin and total protein were determined by the Endospec assay and bicinchoninic acid assay, respectively. MIF was detected in 97.9% of the MEEs from adults, while endotoxin, IL-1beta, TNF-alpha, and RANTES were detected in 96.8, 12.6, 5.3, and 43.9%, respectively. In addition, the level of MIF was significantly higher than those of endotoxin, IL-1beta, and TNF-alpha. A positive correlation between the levels of MIF and endotoxin was observed. MIF and endotoxin were detected in 81.8 and 72.7%, respectively, of the MEEs from the children. The level of MIF was significantly higher in the children, and conversely that of endotoxin was significantly higher in the adults. These results suggest that the interaction between MIF and endotoxin may promote fluid collection in the middle ear, particularly in adults. PMID- 12738642 TI - Recombinant truncated flagellin of Burkholderia pseudomallei as a molecular probe for diagnosis of melioidosis. AB - Current serological tests for melioidosis, using impure or uncharacterized cell antigens from Burkholderia pseudomallei, have problems in detection sensitivity and specificity. Therefore, we designed and expressed the recombinant flagellin (truncated at both the N- and C-terminal ends), and used the antigen to develop an indirect enzyme-linked immunosorbent assay (ELISA) to diagnose melioidosis. Comparison of the immunoreactivities of the full-length and truncated flagellins reveals that the truncated flagellin performed much better in detection specificity and sensitivity. Only the full-length flagellin was recognized by other bacterial causing septicemia and gave a false-positive result in Western analysis, indicating that the cross-reactive epitopes were located on the more highly conserved N- and C-terminal regions of flagellin. The indirect ELISA using recombinant truncated flagellin as the antigen achieved 93.8% sensitivity and 96.3% specificity and offered a more efficient serodiagnosis of melioidosis. PMID- 12738640 TI - Frequency of measles virus-specific CD4+ and CD8+ T cells in subjects seronegative or highly seropositive for measles vaccine. AB - The protective effect of measles immunization is due to humoral and cell-mediated immune responses. Little is known about cell-mediated immunity (CMI) to measles vaccine virus, the relative contribution of CD4(+) and CD8(+) T cells to variability in such immune responses, and the immunologic longevity of the CMI after measles vaccination in humans. Our study characterizes cellular immune response in subjects seronegative or highly seropositive for measles vaccine immunoglobulin G-specific antibody, aged 15 to 25 years, previously immunized with two doses of measles-mumps-rubella II vaccine. We evaluated the ability of subjects to respond to measles vaccine virus by measuring measles virus-specific T-cell proliferation. We examined the frequencies of measles virus-specific memory Th1 and Th2 cells by an ELISPOT assay. Our results demonstrated that proliferation of T cells in seronegative subjects was significantly lower than that for highly seropositive subjects (P = 0.003). Gamma interferon (IFN-gamma) secretion predominated over interleukin 4 (IL-4) secretion in response to measles virus in both groups. The median frequency of measles virus-reactive CD8(+) T cells secreting IFN-gamma was 0.09% in seronegative subjects and 0.43% in highly seropositive subjects (P = 0.04). The median frequency of CD4(+) T cells secreting IL-4 in response to measles virus was 0.03% in seronegative subjects and 0.09% in highly seropositive subjects (P = 0.005). These data confirm the presence of measles virus-specific cellular immune responses post-measles vaccine immunization in humans. The detection of measles virus-induced IFN-gamma and IL-4 production by ELISPOT can be used to identify measles virus-specific low frequency memory T cells in subjects immunized with measles vaccine. These differences agree in directionality with the observed antibody response phenotype. PMID- 12738643 TI - T- and B-cell immune responses of patients who had undergone colectomies to oral administration of Salmonella enterica serovar Typhi Ty21a vaccine. AB - The capacity of an oral live attenuated Salmonella enterica serovar Typhi Ty21a vaccine to induce immune responses in patients who had undergone colectomies because of ulcerative colitis was evaluated, and these responses were compared with those of healthy volunteers. Purified CD4(+) and CD8(+) T cells from peripheral blood were stimulated in vitro by using the heat-killed Ty21a vaccine strain, and the proliferation and gamma interferon (IFN-gamma) production were measured before and 7 or 8 days after vaccination. Salmonella-specific immunoglobulin A (IgA) and IgG antibody responses in serum along with IgA antibody responses in ileostomy fluids from the patients who had undergone colectomies were also evaluated. Three doses of vaccine given 2 days apart failed to induce proliferative T-cell responses in all the six patients who had undergone colectomies, and increases in IFN-gamma production were found only among the CD8(+) cells from three of the patients. In contrast, both proliferative responses and increased IFN-gamma production were observed among CD4(+) and CD8(+) T cells from 3 and 6 of 10 healthy volunteers, respectively. Salmonella-specific IgA and/or IgG antibody responses in serum were observed for five (56%) of nine patients who had undergone colectomies and in 15 (88%) of 17 healthy volunteers. In ileostomy fluids, significant anti-Salmonella IgA antibody titer increases were detected in six (67%) of nine patients who had undergone colectomies. The impaired T- and B-cell immune responses found after vaccination in the circulation of patients who have undergone colectomies may be explained by a diminished colonization of the Ty21a vaccine strain due to the lack of a terminal ileum and colon. PMID- 12738644 TI - Human papillomavirus-specific antibody status in oral fluids modestly reflects serum status in human immunodeficiency virus-positive individuals. AB - Serological assays are valuable tools for studies of the epidemiology of human papillomaviruses (HPVs). The efficacy of a less invasive oral-fluid assay for detection of HPV antibodies was examined. Matched serum, saliva, and oral mucosal transudate (OMT) specimens collected from 150 human immunodeficiency virus seropositive patients were tested for immunoglobulin G antibodies against HPV-6 and HPV-11 combined (HPV-6/11) and HPV-16 capsids. Antibodies to HPV were detected in both types of oral specimens. Seroprevalence rates were 55% for HPV 6/11 and 37% for HPV-16, whereas oral prevalence rates were significantly lower (for HPV-6/11 in saliva, 31%, and in OMT, 19%; for HPV-16 in saliva, 19%, and in OMT, 17%). HPV antibody detection in OMT more accurately reflected the presence of antibodies in serum than did HPV antibody detection in saliva. More stringent saliva assay cutpoints yielded stronger associations between oropositivity and seropositivity; less stringent OMT cutpoints yielded stronger associations between oropositivity and seropositivity. Although HPV antibodies were detected in oral fluids, further optimization of the assay is necessary before oral-fluid testing can be implemented as a reliable alternative to serum testing for HPV. PMID- 12738645 TI - Development of an enzyme-linked immunosorbent assay for immunoglobulin M antibodies against measles virus. AB - Measles is a highly contagious respiratory virus infection, with typical clinical symptoms including maculopapular rash, fever, cough, coryza, and conjunctivitis. Despite implementation of widespread vaccination programs throughout the world, the rates of global morbidity and mortality are still considerable. This study was performed to design a reliable indirect enzyme-linked immunosorbent assay (ELISA) to measure measles-specific immunoglobulin M (IgM). First, human IgM was purified, and then an anti-IgM antibody was produced in rabbits and purified in a multistep process. The rabbit IgG against human IgM was conjugated with peroxidase. Measles virus-infected Vero cells produced viral antigen. One hundred serum samples from infants of 9 to 18 months of age, mostly vaccinated, were evaluated for determining the presence of specific IgM antibodies against measles virus. The samples were also evaluated for neutralizing antibodies against measles virus by a microneutralization test (MNT). By comparing the results of the ELISA with those of MNT, it was demonstrated that ELISA had a sensitivity and specificity of 100 and 92%, respectively. On the other hand, when the results obtained by our ELISA system were compared with those of an imported measles virus IgM ELISA kit (EIAgen; Adaltis Italia SPa, Bologna, Italy), a high level of agreement was shown (k = 0.926). PMID- 12738646 TI - Immunohematological reference ranges for adults from the Central African Republic. AB - A survey was carried out on 150 healthy adults to establish hematological reference ranges for human immunodeficiency virus (HIV)-negative adults from the Central African Republic (CAR). Immunohematological mean values, medians, and 95th-percentile reference ranges were established. Mean values were as follows: leukocyte (WBC) counts, 5.28 x 10(9)/liter (males) and 5.11 x 10(9)/liter (females); erythrocyte counts, 5.20 x 10(12)/liter (males) and 4.50 x 10(12)/liter (females); hemoglobin, 15.1 g/dl (males) and 12.5 g/dl (females); hematocrit, 45% (males) and 37% (females); lymphocytes, 2,587/ micro l (males) and 2,466/ micro l (females); CD4 T cells, 927/ micro l (males) and 940/ micro l (females); CD8 T cells, 898/ micro l (males) and 716/ micro l (females); and CD4/CD8 T-cell ratio, 1.13 (males) and 1.41 (females). We concluded that (i) the WBC and hemoglobin values of healthy HIV-negative adults from the CAR are lower than the reference values currently used in the CAR and (ii) the absolute CD4 T cell counts of healthy HIV-negative adults from the CAR are similar to values for Europeans but the absolute CD8 T-cell counts are much higher. Thus, the CD4/CD8 T cell ratios for healthy adults from the CAR are significantly reduced compared to the ratios for healthy Europeans. PMID- 12738647 TI - Cross-reactivity between Chlamydia trachomatis heat shock protein 10 and early pregnancy factor. AB - Chlamydia trachomatis heat shock protein 10 (Chsp10) is associated with chronic genital tract infection with C. trachomatis. Chsp10 is homologous to human chaperonin 10 (Cpn10) and early pregnancy factor (EPF), a form of human Cpn10 that is specifically secreted at the start of pregnancy. We investigated cross reactions between serum anti-Chsp10 antibodies and anti-EPF antibodies in pregnant and nonpregnant patients. Pregnancy was found to be associated with the presence of anti-EPF antibodies, which are specifically induced in pregnant women with a history of C. trachomatis infection, and with the presence of serum anti Chsp10 antibodies. We also found that infertility was associated with the presence of anti-Chsp10 and anti-EPF antibodies. The HLA class II haplotype DR8 DQ4 was associated with the presence of anti-Chsp10 antibodies but not of anti EPF antibodies. PMID- 12738648 TI - Development of a recombinant protein-based enzyme-linked immunosorbent assay and its applications in field surveillance of rodent mice for presence of immunoglobulin G against Orientia tsutsugamushi. AB - A recombinant protein containing the immunodominant conserved epitope region of the 56-kDa outer membrane protein of the Karp strain of Orientia tsutsugamushi was purified to near homogeneity using recombinant DNA techniques. The purified protein was used to immunize rabbits and produced an antibody that could recognize different strains of O. tsutsugamushi, as demonstrated both by Western blotting and immunofluorescence assay. An enzyme-linked immunosorbent assay (ELISA) based on this recombinant protein was developed to detect antibody (immunoglobulin G [IgG]) against O. tsutsugamushi in mice captured in different districts of Taiwan during 2000 to 2001. A significant difference was found in the antibody seroprevalence rates of Suncus murinus mice captured in different districts of Taiwan (chi(2)(4, 0.95) = 26.64; P < 0.05). Furthermore, a significant difference of IgG seropositivity rates was observed among different kinds of mice (chi(2)(5, 0.95) = 93.85; P < 0.05). Antibody seropositivity rates were higher in Bandicota indica (100%), Rattus flavipectus (96.17%), and Rattus losea (95.83%) than in Rattus norvegicus (86.05%) and Rattus mindanensis (83.67%) (chi(2)(diff, 5, 0.95) = 12.59, P < 0.05). The lowest antibody seropositivity rate (54.4%) was observed in Suncus murinus. Antibody seropositivity rates of mice from different districts differed significantly because of the significant difference in antibody seroprevalence rates for S. murinus. The results of this study indicated that the recombinant protein ELISA developed in this study could be used to conduct large-scale surveillance of rodent mice for the presence of antibody against O. tsutsugamushi. The high seroprevalence rates in rodent mice (except S. murinus) suggest that people residing in these districts are at increased risk of developing O. tsutsugamushi infection. PMID- 12738649 TI - Monoclonal antibodies to Escherichia coli-expressed P46 and P65 membranous proteins for specific immunodetection of Mycoplasma hyopneumoniae in lungs of infected pigs. AB - The P46 and P65 proteins of Mycoplasma hyopneumoniae are two membranous proteins carrying species-specific antigenic determinants. Based on the genomic sequence of the reference strain ATCC 25934, primers were designed for PCR amplification of the genes encoding entire P46 (1,260 bp) and P65 (1,803 bp) and N-terminally truncated P65(c) (1,200 bp). These primers were shown to be specific to M. hyopneumoniae since no DNA amplicons could be obtained with other mycoplasma species that commonly colonize the porcine respiratory tract. Both amplified genes were then cloned into the pGEX-4T-1 vector to be expressed in Escherichia coli cells as recombinant fusion proteins with glutathione S-transferase (GST). Prior to generation of expression constructs, TGA nonsense codons, exceptionally used for tryptophan residues by M. hyopneumoniae, had been converted to TGG codons by PCR-directed mutagenesis. Following induction by IPTG (isopropyl-beta-D thiogalactopyranoside), both GST-P46 and GST-P65(c) recombinant fusion proteins were recovered by disrupting transformed cells by sonication, purified by affinity chromatography, and then cut with thrombin to release the P46 and P65(c) moieties. The enriched E. coli-expressed P46 and P65c proteins were used to immunize female BALB/c mice for the generation of anti-P46 and anti-P65(c) monoclonal antibodies (MAbs). The polypeptide specificities of MAbs obtained was confirmed by Western blotting with cell lysates prepared from the homologous strain. Cross-reactivity study of the anti-P46 and anti-P65(c) MAbs towards two other M. hyopneumoniae reference strains (ATCC 25095 and J strains) and Quebec field strains that had been isolated in culture, suggested that the MAbs obtained against both membranous proteins were directed against highly conserved species specific epitopes. No reactivity to other mycoplasma species tested was demonstrated. Clinical signs and lesions suggestive of enzootic pneumonia were reproduced in specific-pathogen-free pigs that had been inoculated intratracheally with a virulent Quebec field strain (IAF-DM9827) of M. hyopneumoniae. Both anti-P46 and anti-P65(c) MAbs permitted effective detection by indirect immunofluorescence and indirect immunoperoxidase assay of M. hyopneumoniae in, respectively, frozen and formalin-fixed, paraffin-embedded lung sections from pigs that were killed after the 6- to 7-week observation period. PMID- 12738650 TI - Effect of glycosphingolipids purified from Leishmania (Leishmania) amazonensis amastigotes on human peripheral lymphocytes. AB - The effect of purified glycosphingolipids from Leishmania (Leishmania) amazonensis on human lymphoproliferation, on expression of human lymphocyte and monocyte markers (CD3, CD4, CD8, CD14, CD19, and CD45), and on lymphocyte protein kinase C activity was analyzed. PMID- 12738651 TI - Bactericidal/permeability-increasing protein is expressed by human dermal fibroblasts and upregulated by interleukin 4. AB - The bactericidal/permeability-increasing protein (BPI) is an antibiotic- and endotoxin-neutralizing protein of granulocytes and epithelial cells. Constitutive expression of BPI, which increases upon interleukin 4 stimulation, by human dermal fibroblast was demonstrated, suggesting an important role of BPI in gram negative bacterial clearance and a dampened response to endotoxin in the skin. PMID- 12738652 TI - One-step immunochromatographic dipstick tests for rapid detection of Vibrio cholerae O1 and O139 in stool samples. AB - We describe the development and evaluation of a rapid diagnostic test for Vibrio cholerae O1 and O139 based on lipopolysaccharide detection using gold particles. The specificity ranged between 84 and 100%. The sensitivity of the dipsticks ranged from 94.2 to 100% when evaluated with stool samples obtained in Madagascar and Bangladesh. The dipstick can provide a simple tool for epidemiological surveys. PMID- 12738653 TI - Lymphocyte transformation test for medicinal herbs yields false-positive results for first-visit patients. AB - We performed lymphocyte transformation tests (LTTs) for toki (angelicae radix) and ogon (scutellariae radix) on first-visit patients who had never taken Kampo medicines. LTTs for both herbs were positive in 12 of 14 patients, suggesting that LTTs for these herbs are unreliable for the diagnosis of Kampo medicine induced liver injury. PMID- 12738654 TI - Comparison of complement fixation and hemagglutination inhibition assays for detecting antibody responses following influenza virus vaccination. AB - Complement fixation (CF) was compared to hemagglutination inhibition (HI) as a method for identifying antibody responses to influenza virus vaccination. CF assays were performed at two different laboratories using paired (pre- and postvaccination) sera from 38 vaccinated laboratory employees; HI assays were performed at a third laboratory. As expected, most vaccinees (31/38 = 82%) responded to at least one of three influenza virus antigens as measured by HI. In contrast, only 21% (8/38) of vaccinees showed a response by CF at laboratory 1, and only 29% (11/38) showed a response by CF at laboratory 2. These findings indicate that due to low sensitivity, CF assays should not be used to assess the antibody response to influenza virus vaccination. PMID- 12738655 TI - Serological survey of Toscana virus infections in a high-risk population in Italy. AB - Toscana virus is the most important agent responsible for meningitis in central Italy. We report a serosurveillance study, using an immunoenzymatic assay, of 360 serum samples harvested from a high-risk population occupationally exposed to Toscana virus in two regions of Italy, Tuscany and Piedmont. The results indicates a seroprevalence of Toscana virus of 77.2% in the forestry workers, particularly in the Tuscany region. This fact is strictly correlated with the ecological niches specific for the survival of Toscana virus arthropod vector. PMID- 12738656 TI - Peptidoglycan induces mobilization of the surface marker for activation marker CD66b in human neutrophils but not in eosinophils. AB - Peptidoglycan from Staphylococcus aureus mobilized CD66b in human neutrophils but did not upregulate surface activation markers in eosinophils. In addition, Toll like receptor 2, implicated in the recognition of peptidoglycan, was detected on the surface of resting neutrophils but not on eosinophils. These findings suggest roles for neutrophils but not eosinophils in innate recognition of peptidoglycan. PMID- 12738657 TI - A patient with Crimean-Congo hemorrhagic fever serologically diagnosed by recombinant nucleoprotein-based antibody detection systems. AB - We treated a male patient with Crimean-Congo hemorrhagic fever (CCHF). The diagnosis of CCHF was confirmed by reverse transcription-PCR and recombinant nucleoprotein (rNP)-based immunoglobulin G (IgG) and IgM capture enzyme-linked immunosorbent assays of serially collected serum samples. The patient was treated with intravenous ribavirin and recovered with no consequences. The study indicates that rNP-based CCHF virus antibody detection systems are useful for confirming CCHF virus infections. This case also suggests that intravenous ribavirin therapy may be promising for the treatment of CCHF patients. PMID- 12738658 TI - Anti-hepatitis A virus immunoglobulin M antibodies in urine samples for rapid diagnosis of outbreaks. AB - The main goal of this study was to test the feasibility of using urine for diagnosing hepatitis A virus (HAV) infections. A correlation of 90.78% between the test results of urine and serum samples was obtained. Four outbreaks of hepatitis A were confirmed by testing only urine samples. The levels of anti-HAV immunoglobulin M (IgM) antibodies in urine samples remained stable during 6 months of storage at -70 degrees C but decreased when the samples were stored at 4 degrees C. The results of tests of samples obtained 2 and 6 months after infection suggested that IgM levels decline more rapidly in urine than in serum. PMID- 12738659 TI - Activation of coagulation by administration of recombinant factor VIIa elicits interleukin 6 (IL-6) and IL-8 release in healthy human subjects. AB - The activation of coagulation has been shown to contribute to proinflammatory responses in animal and in vitro experiments. Here we report that the activation of coagulation in healthy human subjects by the administration of recombinant factor VIIa also elicits a small but significant increase in the concentrations of interleukin 6 (IL-6) and IL-8 in plasma. This increase was absent when the subjects were pretreated with recombinant nematode anticoagulant protein c2, the inhibitor of tissue factor-factor VIIa. PMID- 12738660 TI - Identification of a gene expression signature associated with pediatric AML prognosis. AB - Most patients with acute myeloid leukemia (AML) enter complete remission (CR) after treatment with chemotherapy, but a large number of them experience relapse with resistant disease. To identify genes that are associated with their prognoses, we analyzed gene expression in 54 pediatric patients with AML using an oligonucleotide microarray that contained 12 566 probe sets. A supervised approach using the Student t test selected a prognostic set of 35 genes, some of which are associated with the regulation of cell cycle and apoptosis. Most of these genes had not previously been reported to be associated with prognosis and were not correlated with morphologically classified French-American-British (FAB) subtypes or with karyotypes. These results indicate the existence of prognosis associated genes that are independent of cell lineage and cytogenetic abnormalities, and they can provide therapeutic direction for individual risk adapted therapy for pediatric AML patients. PMID- 12738661 TI - Hair follicles serve as local reservoirs of skin mast cell precursors. AB - Several leukocyte populations normally reside in mouse skin, including Langerhans cells and gammadelta T cells in the epidermis and macrophage and mast cells in the dermis. Interestingly, these skin resident leukocytes are frequently identified within or around hair follicles (HFs), which are known to contain stem cell populations that can generate the epidermal architecture or give rise to the melanocyte lineage. Thus, we reasoned that HFs might serve as a local reservoir of the resident leukocyte populations in the skin. When vibrissal follicles of adult mice were cultured in the presence of stem cell factor (SCF), interleukin 3 (IL-3), IL-7, granulocyte-macrophage colony-stimulating factor, and Flt3 ligand, CD45+/lineage-/c-kit+/FcepsilonRI+ cells became detectable on the outgrowing fibroblasts in 10 days and expanded progressively thereafter. These HF-derived leukocytes showed characteristic features of connective tissue-type mast cells, including proliferative responsiveness to SCF, metachromatic granules, mRNA expression for mast cell proteases-1, -4, -5, and -6, and histamine release on ligation of surface IgE or stimulation with substance P or compound 48/80. These results, together with our findings that HFs contain c-kit+ cells and produce SCF mRNA and protein, suggest that HFs provide a unique microenvironment for local development of mast cells. PMID- 12738662 TI - Granule stores from cellubrevin/VAMP-3 null mouse platelets exhibit normal stimulus-induced release. AB - It is widely accepted that the platelet release reaction is mediated by heterotrimeric complexes of integral membrane proteins known as SNAREs (SNAP receptors). In an effort to define the precise molecular machinery required for platelet exocytosis, we have analyzed platelets from cellubrevin/VAMP-3 knockout mice. Cellubrevin/VAMP-3 has been proposed to be a critical v-SNARE for human platelet exocytosis; however, data reported here suggest that it is not required for platelet function. Upon stimulation with increasing concentrations of thrombin, collagen, or with thrombin for increasing time there were no differences in secretion of [3H]-5HT (dense core granules), platelet factor IV (alpha granules), or hexosaminidase (lysosomes) between null and wild-type platelets. There were no gross differences in bleeding times nor in agonist induced aggregation measured in platelet-rich plasma or with washed platelets. Western blotting of wild-type, heterozygous, and null platelets confirmed the lack of cellubrevin/VAMP-3 in nulls and showed that most elements of the secretion machinery are expressed at similar levels. While the secretory machinery in mice was similar to humans, mice did express apparently higher levels of synaptobrevin/VAMP-2. These data show that the v-SNARE, cellubrevin/VAMP-3 is not a requirement for the platelet release reaction in mice. PMID- 12738663 TI - Prior gemtuzumab ozogamicin exposure significantly increases the risk of veno occlusive disease in patients who undergo myeloablative allogeneic stem cell transplantation. AB - Gemtuzumab ozogamicin (GO), a monoclonal antibody used in the treatment of acute myelogenous leukemia (AML) has been linked to the development of venoocclusive disease (VOD). We conducted a retrospective study of 62 patients with previously treated AML/MDS (myelodysplastic syndrome) who underwent allogeneic stem cell (SC) transplantation at our institution from December 2000 to October 2002 to determine whether GO exposure prior to allogeneic SC transplantation increases the risk of developing VOD. Fourteen patients received GO prior to SC transplantation. Of 62 patients, 13 (21%) developed VOD; 9 (64%) of 14 with prior GO exposure developed VOD compared with 4 (8%) of 48 without prior GO exposure (P <.0001). Logistic regression controlling for sex, disease status, donor type, and graft-versus-host disease prophylaxis identified prior treatment with GO as a significant risk factor for VOD (odds ratio [OR], 21.6; 95% confidence interval [CI], 4.2-112.2]. Nine of 10 patients who underwent SC transplantation 3.5 months or less following GO developed VOD compared with none of 4 patients who underwent SC transplantation more than 3.5 months from GO administration. Three of 14 patients who received GO prior to SC transplantation died of VOD. We conclude that patients undergoing SC transplantation within a short interval from GO administration are at increased risk of developing VOD. PMID- 12738664 TI - In vitro generation of T lymphocytes from embryonic stem cell-derived prehematopoietic progenitors. AB - Embryonic stem (ES) cells can differentiate into most blood cells in vitro, providing a powerful model system to study hematopoiesis. However, ES cell derived T lymphocytes have not been generated in vitro, and it was unresolved whether such potential is absent or merely difficult to isolate. Because the latter case might result from rapid commitment to non-T-cell fates, we isolated ES cell-derived prehematopoietic precursors for reconstitution of fetal thymic organ cultures. We found a transient Flk1+CD45- subset of these precursors generated T lymphocytes in vitro, and the use of reaggregate thymic organ cultures greatly enhanced reconstitution frequency. These findings reveal that ES cells can exhibit in vitro T-cell potential, but this is restricted to early stages of ES cell differentiation. Moreover, the results support the notion that the thymic microenvironment can induce T-cell differentiation from a subset of prehematopoietic progenitors and suggest deficient migration into intact thymi hindered previous attempts to generate T cells in vitro from ES cell-derived progenitors. These findings demonstrate that a defined subset of ES cells has the potential to generate T cells in vitro and could contribute to greater understanding of the molecular events of hematopoietic induction and T-cell lineage commitment. PMID- 12738665 TI - Hematopoietic stimulation by a dipeptidyl peptidase inhibitor reveals a novel regulatory mechanism and therapeutic treatment for blood cell deficiencies. AB - In hematopoiesis, cytokine levels modulate blood cell replacement, self-renewal of stem cells, and responses to disease. Feedback pathways regulating cytokine levels and targets for therapeutic intervention remain to be determined. Amino boronic dipeptides are orally bioavailable inhibitors of dipeptidyl peptidases. Here we show that the high-affinity inhibitor Val-boro-Pro (PT-100) can stimulate the growth of hematopoietic progenitor cells in vivo and can accelerate neutrophil and erythrocyte regeneration in mouse models of neutropenia and acute anemia. Hematopoietic stimulation by PT-100 correlated with increased cytokine levels in vivo. In vitro, PT-100 promoted the growth of primitive hematopoietic progenitor cells by increasing granulocyte-colony-stimulating factor (G-CSF), interleukin-6 (IL-6), and IL-11 production by bone marrow stromal cells. Two molecular targets of PT-100 are expressed by stromal cells- CD26/DPP-IV and the closely related fibroblast activation protein (FAP). Because PT-100 was active in the absence of CD26, FAP appears to be the hematopoietic target for PT-100. Interaction of PT-100 with the catalytic site seems to be required because amino terminal acetylation of PT-100 abrogated enzyme inhibition and hematopoietic stimulation. PT-100 is a therapeutic candidate for the treatment of neutropenia and anemia. The data support increasing evidence that dipeptidyl peptidases can regulate complex biologic systems by the proteolysis of signaling peptides. PMID- 12738666 TI - Molecular remission after myeloablative allogeneic stem cell transplantation predicts a better relapse-free survival in patients with multiple myeloma. AB - Patients in complete clinical remission after myeloablative allogeneic stem cell transplantation (allo-SCT) were enrolled in a longitudinal study to assess the predictive value of molecular monitoring. Using polymerase chain reaction (PCR) for immunoglobulin gene rearrangements it was possible to generate a clone specific molecular marker in 48 of 70 patients. Of these 48 patients, 16 (33%) attained durable PCR-negativity after transplantation, whereas 13 (27%) remained persistently PCR-positive and 19 (40%) showed a mixed pattern. The cumulative risk of relapse at 5 years was 0% for PCR-negative patients, 33% for PCR-mixed patients, and 100% for PCR-positive patients. Within the group studied it was not possible to identify any clinical feature predictive of durable PCR-negativity. We believe that these findings could prompt the design of prospective studies to evaluate if the treatment of molecular disease can extend remission duration and survival. PMID- 12738667 TI - Heat shock protein 70 binds caspase-activated DNase and enhances its activity in TCR-stimulated T cells. AB - DNA fragmentation is a hallmark of cells undergoing apoptosis and is mediated mainly by the caspase-activated DNase (CAD or DNA-fragmentation factor 40 [DFF40]), which is activated when released from its inhibitor protein (ICAD or DFF45) upon apoptosis signals. Here we analyzed the effect of heat shock protein 70 (Hsp70) on CAD activity in T-cell receptor (TCR)-induced apoptosis using a T cell line (TAg-Jurkat). Overexpression of Hsp70 significantly augmented the apoptotic cell death as well as DNA fragmentation in CD3/CD28- or staurosporine stimulated cells. Following stimulation of cells with CD3/CD28 or staurosporine, Hsp70 was coprecipitated with free CAD, but not with CAD associated with ICAD. Furthermore, the purified Hsp70 dose-dependently augmented DNA-fragmentation activity of caspase-3-activated CAD in a cell-free system. Peptide-binding domain deleted Hsp70 could neither bind nor augment its activity, while adenosine triphosphate (ATP)-binding domain-deleted Hsp70 or the peptide-binding domain itself bound CAD and augmented its activity. These results indicate that the the binding of Hsp70 to the activated CAD via the peptide-binding domain augments its activity. Although CAD lost its activity in an hour after being released from ICAD in vitro, its activity was retained after an hour of incubation in the presence of Hsp70, suggesting that Hsp70 may be involved in stabilization of CAD activity. Finally, CAD that had been coprecipitated with Hsp70 from the cell lysate of staurosporine-activated 293T cells induced chromatin DNA fragmentation and its activity was not inhibited by ICAD. These results suggest that Hsp70 binds free CAD in TCR-stimulated T cells to stabilize and augment its activity. PMID- 12738668 TI - Drug-induced thrombocytopenia: localization of the binding site of GPIX-specific quinine-dependent antibodies. AB - Immune thrombocytopenia is a common complication of therapy with a large number of drugs. The most widely studied drug-induced immune thrombocytopenia (DIT) is caused by quinine. In most cases of DIT, antibodies bind to the platelet membrane glycoprotein (GP) Ib-IX complex in a drug-dependent fashion and bring about increased platelet clearance by the reticuloendothelial system resulting in thrombocytopenia. Here, we report the characterization of the quinine-dependent antibody activity of sera from 13 patients with quinine-induced thrombocytopenia. In our series of patients, GPIX was the most prevalent target of quinine dependent antibodies. To identify the structural determinants of GPIX recognized by quinine-dependent antibodies, 4 chimeric mouse/human GPIX constructs and stable Chinese hamster ovary (CHO) cell lines that expressed the chimeras in association with GPIbalpha and GPIbbeta were produced. The analysis of 6 patient sera with the chimeric cell lines provided evidence for localization of the anti GPIX quinine-dependent antibody binding site to the C-ext region (amino acid [aa] 64-135) of human GPIX. Further characterization of the C-ext region of the GPIX indicated that replacement of the Arg110 and Gln115 of the human GPIX with the corresponding residues from mouse (Gln and Glu, respectively) resulted in a significant reduction in the binding of GPIX antibodies in our series of patients, with Arg110Gln, giving a more pronounced effect than Gln115Glu. Hence, these 2 residues, particularly Arg110, play an important role in the structure of the antigenic site on GPIX recognized by anti-GPIX antibodies. PMID- 12738669 TI - The contribution of glycoprotein VI to stable platelet adhesion and thrombus formation illustrated by targeted gene deletion. AB - Platelet interaction with exposed adhesive ligands at sites of vascular injury is required to initiate a normal hemostatic response and may become a pathogenic factor in arterial diseases leading to thrombosis. We report a targeted disruption in a key receptor for collagen-induced platelet activation, glycoprotein (GP) VI. The breeding of mice with heterozygous GP VI alleles produced the expected frequency of wild-type, heterozygous, and homozygous genotypes, indicating that these animals had no reproductive problems and normal viability. GP VInull platelets failed to aggregate in response to type I fibrillar collagen or convulxin, a snake venom protein and known platelet agonist of GP VI. Nevertheless, tail bleeding time measurements revealed no severe bleeding tendency as a consequence of GP VI deficiency. Ex vivo platelet thrombus formation on type I collagen fibrils was abolished using blood from either GP VInull or FcR-gammanull animals. Reflection interference contrast microscopy revealed that the lack of thrombus formation by GP VInull platelets could be linked to a defective platelet activation following normal initial tethering to the surface, visualized as lack of spreading and less stable adhesion. These results illustrate the role of GP VI in postadhesion events leading to the development of platelet thrombi on collagen fibrils. PMID- 12738670 TI - Sustained phenotypic correction of canine hemophilia A using an adeno-associated viral vector. AB - Gene therapy for hemophilia A requires efficient delivery of the factor VIII gene and sustained protein expression at circulating levels of at least 1% to 2% of normal. Adeno-associated viral type 2 (AAV2) vectors have a number of advantages over other viral vectors, including an excellent safety profile and persistent gene expression. However, a major disadvantage is their small packaging capacity, which has hampered their use in treating diseases such as hemophilia A, cystic fibrosis, and muscular dystrophy, which are caused by mutations in large genes. Here we demonstrate that this can be overcome by using small regulatory elements to drive expression of a B-domain-deleted form of FVIII. The use of this vector for hepatic gene transfer in a canine model of hemophilia A resulted in the sustained (> 14 months) expression of biologically active FVIII. FVIII activity levels of 2% to 4% were achieved. These levels correlated with a partial correction in the whole-blood clotting time and cuticle bleeding time. In addition, immunoprecipitation analysis demonstrated the expression of canine FVIII of the predicted size in the plasma of injected animals. These data support the use of AAV2 vectors in human clinical trials to treat hemophilia A patients. PMID- 12738672 TI - Antiapoptotic effect of coagulation factor VIIa. AB - Binding of factor VIIa (FVIIa) to its cellular receptor tissue factor (TF) was previously shown to induce various intracellular signaling events, which were thought to be responsible for TF-mediated biologic effects, including angiogenesis, tumor metastasis, and restenosis. To understand the mechanisms behind these processes, we have examined the effect of FVIIa on apoptosis. Serum deprivation-induced apoptosis of BHK(+TF) cells was characterized by apoptotic blebs, nuclei with chromatin-condensed bodies, DNA degradation, and activation of caspase 3. FVIIa markedly decreased the number of cells with apoptotic morphology and prevented the DNA degradation as measured by means of TdT-mediated dUTP nick end labeling (TUNEL). The antiapoptotic effect of FVIIa was confirmed by the observation that FVIIa attenuated caspase 3 activation. FVIIa-induced antiapoptotic effect was dependent on its proteolytic activity and TF but independent of factor Xa and thrombin. FVIIa-induced cell survival correlated with the activation of Akt and was inhibited markedly by the specific PI3-kinase inhibitor, LY294002. Blocking the activation of p44/42 mitogen-activated protein kinase (MAPK) by the specific mitogen-induced extracellular kinase (MEK) inhibitor, U0126, impaired modestly the ability of FVIIa to promote cell survival. In conclusion, FVIIa binding to TF provided protection against apoptosis induced by growth factor deprivation, primarily through activation of PI3-kinase/Akt pathway, and to a lesser extent, p44/42 MAPK pathway. PMID- 12738671 TI - A phase 2 trial of CHOP chemotherapy followed by tositumomab/iodine I 131 tositumomab for previously untreated follicular non-Hodgkin lymphoma: Southwest Oncology Group Protocol S9911. AB - Advanced follicular lymphoma is incurable with conventional chemotherapy and radiotherapy. The Southwest Oncology Group (SWOG) conducted a phase 2 trial (S9911) of a novel regimen consisting of 6 cycles of CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone) chemotherapy followed 4 to 8 weeks later by tositumomab/iodine I 131 tositumomab (anti-CD20 antibody) in 90 eligible patients with previously untreated, advanced stage follicular lymphoma. Treatment was well tolerated. Reversible myelosuppression was the main adverse event and was more severe during CHOP chemotherapy than following radioimmunotherapy. The overall response rate to the entire treatment regimen was 90%, including 67% complete remissions (CRs plus unconfirmed CRs [CRu's]) and 23% partial remissions (PRs). Twenty-seven (57%) of the 47 fully evaluable patients who achieved less than a CR with CHOP improved their remission status after tositumomab/iodine I 131 tositumomab. With a median follow-up of 2.3 years, the 2-year progression free survival (PFS) was estimated to be 81%, with a 2-year overall survival of 97%. This study has established the feasibility, tolerability, and efficacy of this regimen for patients with advanced follicular lymphoma. This novel treatment appears promising compared with the SWOG's historical experience using CHOP alone and is currently being compared with CHOP plus rituximab in a randomized phase 3 trial (S0016). PMID- 12738673 TI - Functional comparison of DCs generated in vivo with Flt3 ligand or in vitro from blood monocytes: differential regulation of function by specific classes of physiologic stimuli. AB - Dendritic cells (DCs) are a family of leukocytes that initiate T- and B-cell immunity against pathogens. Migration of antigen-loaded DCs from sites of infection into draining lymphoid tissues is fundamental to the priming of T-cell immune responses. In humans, the major peripheral blood DC (PBDC) types, CD1c+ DCs and interleukin 3 receptor-positive (IL-3R+) plasmacytoid DCs, are significantly expanded in vivo with the use of Flt3 ligand (FL). DC-like cells can also be generated from monocyte precursors (MoDCs). A detailed comparison of the functional potential of these types of DCs (in an autologous setting) has yet to be reported. Here, we compared the functional capacity of FL-expanded CD1c+ PBDCs with autologous MoDCs in response to 3 different classes of stimuli: (1) proinflammatory mediators, (2) soluble CD40 ligand trimer (CD40L), and (3) intact bacteria (Escherichia coli). Significant differences in functional capacities were found with respect to changes in phenotype, migratory capacity, cytokine secretion, and T-cell stimulation. MoDCs required specific stimuli for the expression of functions. They responded vigorously to CD40L or E coli, expressing cytokines known to regulate interferon-gamma (IFN-gamma) in T cells (IL-12p70, IL 18, and IL-23), but required prostaglandin E2 (PGE2) during stimulation to migrate to chemokines. In contrast, PBDCs matured in response to minimal stimulation, rapidly acquired migratory function in the absence of PGE2 containing stimuli, and were low cytokine producers. Interestingly, both types of DCs were equivalent with respect to stimulation of allogeneic T-cell proliferation and presentation of peptides to cytotoxic T lymphocyte (CTL) lines. These distinct differences are of particular importance when considering the choice of DC types for clinical applications. PMID- 12738674 TI - Synergistic antileukemic interactions between 17-AAG and UCN-01 involve interruption of RAF/MEK- and AKT-related pathways. AB - Interactions between the protein kinase C (PKC) and Chk1 inhibitor UCN-01 and the heat shock protein 90 (Hsp90) antagonist 17-AAG have been examined in human leukemia cells in relation to effects on signal transduction pathways and apoptosis. Simultaneous exposure (30 hours) of U937 monocytic leukemia cells to minimally toxic concentrations of 17-AAG (eg, 400 nM) and UCN-01 (eg, 75 nM) triggered a pronounced increase in mitochondrial injury (ie, loss of mitochondrial membrane potential [Deltapsim]; cytosolic release of cytochrome c), caspase activation, and apoptosis. Synergistic induction of apoptosis was also observed in other human leukemia cell types (eg, Jurkat, NB4). Coexposure of human leukemia cells to 17-AAG and the PKC inhibitor bisindolylmaleimide (GFX) did not result in enhanced lethality, arguing against the possibility that the PKC inhibitory actions of UCN-01 are responsible for synergistic interactions. The enhanced cytotoxicity of this combination was associated with diminished Akt activation and marked down-regulation of Raf-1, MEK1/2, and mitogen-activated protein kinase (MAPK). Coadministration of 17-AAG and UCN-01 did not modify expression of Hsp90, Hsp27, phospho-JNK, or phospho-p38 MAPK, but was associated with further p34cdc2 dephosphorylation and diminished expression of Bcl-2, Mcl-1, and XIAP. In addition, inducible expression of both a constitutively active MEK1/2 or myristolated Akt construct, which overcame inhibition of ERK and Akt activation, respectively, significantly attenuated 17-AAG/UCN-01-mediated lethality. Together, these findings indicate that the Hsp90 antagonist 17-AAG potentiates UCN-01 cytotoxicity in a variety of human leukemia cell types and suggest that interference with both the Akt and Raf-1/MEK/MAP kinase cytoprotective signaling pathways contribute to this phenomenon. PMID- 12738675 TI - Induction of FucT-VII by the Ras/MAP kinase cascade in Jurkat T cells. AB - Induction of the alpha1,3-fucosyltransferase FucT-VII in T lymphocytes is crucial for selectin ligand formation, but the signaling and transcriptional pathways that govern FucT-VII expression are unknown. Here, using a novel, highly phorbol myristate acetate (PMA)-responsive variant of the Jurkat T-cell line, we identify Ras and downstream mitogen-activated protein (MAP) kinase pathways as essential mediators of FucT-VII gene expression. PMA induced FucT-VII in only a subset of treated cells, similar to expression of FucT-VII in normal activated CD4 T cells. Introduction of constitutively active Ras or Raf by recombinant retroviruses induced FucT-VII expression only in that subset of cells expressing the highest levels of Ras, suggesting that induction of FucT-VII required a critical threshhold of Ras signaling. Both PMA treatment and introduction of active Ras led to rolling on E-selectin. Pharmacologic inhibition studies confirmed the involvement of the classic Ras-Raf-MEK-extracellular signal-regulated kinase (Ras Raf-MEK-ERK) pathway in FucT-VII induction by PMA, Ras, and Raf. These studies also revealed a second, Ras-induced, Raf-1-independent pathway that participated in induction of FucT-VII. Strong activation of Ras represents a major pathway for induction of FucT-VII gene expression in T cells. PMID- 12738676 TI - Rapid and complete donor chimerism in adult recipients of unrelated donor umbilical cord blood transplantation after reduced-intensity conditioning. AB - Reduced-intensity conditioning may reduce transplantation-related mortality in high-risk adults undergoing hematopoietic transplantation. We investigated unrelated donor umbilical cord blood (UCB) transplantation after such conditioning in 43 patients (median age, 49.5 years; range, 22-65 years) with a primary end point of donor engraftment. The first 21 patients received busulfan 8 mg/kg, fludarabine 200 mg/m2, and 200 cGy of total body irradiation (Bu/Flu/TBI). Subsequent patients (n = 22) received cyclophosphamide 50 mg/kg, fludarabine 200 mg/m2, and 200 cGy TBI (Cy/Flu/TBI). UCB grafts (93%) were 1-2 HLA antigen mismatched with the recipient and contained a median cryopreserved cell dose of 3.7 x 107 (range, 1.6 x 107-6.0 x 107) nucleated cells per kilogram of recipient body weight (NC/kg). Graft versus host disease (GVHD) prophylaxis was cyclosporin A to day 180 plus mycophenolate mofetil to day 30. The cumulative incidence of sustained donor engraftment was 76% (95% confidence interval [CI], 56%-96%) for Bu/Flu/TBI recipients and 94% (95% CI, 84%-100%) for Cy/Flu/TBI recipients. The median day of neutrophil recovery (at least 0.5 x 109/L) for engrafting Bu/Flu/TBI recipients was 26 days (range, 12-30 days) and for Cy/Flu/TBI recipients was 9.5 days (range, 5-28 days). Incidence of grades III-IV acute GVHD was 9% (95% CI, 1%-17%), and survival at 1 year was 39% (95% CI, 23%-56%). These data demonstrate that 0-2 antigen mismatched UCB is sufficient to engraft most adults after reduced-intensity conditioning and is associated with a low incidence of severe acute GVHD. PMID- 12738677 TI - Contribution of beta-2 microglobulin levels to the prognostic stratification of survival in patients with myelodysplastic syndrome (MDS). AB - Prospective analysis of the importance of the plasma levels of beta-2 microglobulin (B2M) in 553 patients with myelodysplastic syndrome (MDS) found that B2M is an independent prognostic variable for survival with weighted significance second only to the karyotype. The incorporation of the B2M covariate into risk assessment of MDS patients added significantly to the power of the IPSS to stratify MDS patients into risk categories. Our results further document that the 2 objectively measured covariates that display the highest power to predict survival, that is, karyotype and B2M, can alone be used for risk stratification. While the results must be verified in an independent and comparable population, our data strongly recommend routine measurement of B2M in patients with MDS. PMID- 12738678 TI - Mithramycin induces fetal hemoglobin production in normal and thalassemic human erythroid precursor cells. AB - We report in this paper that the DNA-binding drug mithramycin is a potent inducer of gamma-globin mRNA accumulation and fetal hemoglobin (HbF) production in erythroid cells from healthy human subjects and beta-thalassemia patients. Erythroid precursors derived from peripheral blood were grown in 2-phase liquid culture. In this procedure, early erythroid progenitors proliferate and differentiate during phase 1 (in the absence of erythropoietin) into late progenitors. In phase 2, in the presence of erythropoietin, the latter cells continue their proliferation and mature into Hb-containing orthochromatic normoblasts. Compounds were added on days 4 to 5 of phase 2 (when cells started to synthesize Hb), and cells were harvested on day 12. Accumulation of mRNAs for gamma-globin, beta-globin, alpha-globin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and beta-actin were measured by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR); induction of HbF was analyzed by high-performance liquid chromatography (HPLC) and, at cellular level, by flow cytometry. We demonstrated that mithramycin was able to up-regulate preferentially gamma-globin mRNA production and to increase HbF accumulation, the percentage of HbF-containing cells, and their HbF content. Mithramycin was more effective than hydroxyurea, being, in addition, not cytotoxic. This was shown by the lack of cytotoxicity on erythroid and myeloid in vitro primary cell cultures treated with mithramycin at concentrations effective for HbF induction. These results are of potential clinical significance because an increase of HbF alleviates the symptoms underlying beta-thalassemia and sickle cell anemia. The results of this report suggest that mithramycin and its analogs warrant further evaluation as potential therapeutic drugs. PMID- 12738679 TI - Platelet alpha2beta1 integrin activation: contribution of ligand internalization and the alpha2-cytoplasmic domain. AB - The alpha2beta1 integrin is a major collagen receptor on platelets. Although it has been proposed that alpha2beta1, like alphaIIbbeta3, undergoes agonist-induced activation, neither the potential contributions of alpha2beta1 receptor/ligand internalization to the increase in ligand binding nor the roles of the alpha2 and beta1 cytoplasmic domains in activation of this integrin have been previously explored. Activation of alpha2beta1 was assessed with fluorescein isothiocyanate labeled soluble type I collagen binding to platelets by flow cytometry. Although collagen internalization in response to agonist activation of platelets was significant, agonist-induced collagen binding still occurred under conditions that block internalization, with minimal changes in cell surface alpha2beta1 expression. Introduction of cell-permeable peptides containing the alpha2 cytoplasmic tail, and especially the membrane proximal KLGFFKR domain, induced alpha2beta1 activation in resting platelets, whereas a cell-permeable peptide containing the beta1 cytoplasmic tail was without effect. Thus, collagen binding to stimulated platelets is increased due to alpha2beta1 activation, in addition to internalization, and the GFFKR motif appears to play an important role in the activation process. PMID- 12738680 TI - BCL6 gene translocation in follicular lymphoma: a harbinger of eventual transformation to diffuse aggressive lymphoma. AB - Follicular lymphoma (FL) is characterized by a relatively indolent clinical course, but the disease often transforms into a more aggressive large cell lymphoma with a rapidly progressive clinical course. In the present study, we analyzed 41 cases of FL known to have subsequently transformed to aggressive lymphoma and an additional 64 FL samples from patients not subsequently transformed. We studied BCL6 gene rearrangement by the methodology of long distance inverse polymerase chain reaction (LDI-PCR). Of the 41 cases known to transform, 16 (39.0%) harbored BCL6 translocation or deletion at the time of FL diagnosis. Among 64 cases not known to transform, BCL6 translocation was detected in 9 (14.1%). The prevalence of BCL6 translocation in the group known to transform was significantly higher (P =.0048). Among the transformation cases, the partners of the BCL6 translocation were identified in 13 cases and included IGH, CIITA, U50HG, MBNL, GRHPR, LRMP, EIF4A2, RhoH/TTF, and LOC92656 (similar to NAPA), whereas in the control group the BCL6 partners were IGH, CIITA, SIAT1, and MBNL. In 13 cases paired specimens before and after transformation were available. Among these paired specimens, a loss (3 cases) or a gain (1 case) of BCL6 translocation was observed after the transformation. Analysis of clonality showed that all of these cases represented the evolution of a subclone of the original tumor population. Our study demonstrated that BCL6 translocation is not necessary for transformation but that BCL6 translocation in FL may constitute a subgroup with a higher risk to transform into aggressive lymphoma. PMID- 12738681 TI - Regulation and mechanisms of ATP-binding cassette transporter A1-mediated cellular cholesterol efflux. AB - ATP-binding cassette transporter A1 (ABCA1) plays a major role in cholesterol homeostasis and HDL metabolism. ABCA1 mediates cellular cholesterol and phospholipid efflux to lipid-poor apolipoproteins, and upregulation of ABCA1 activity is antiatherogenic. ApoA-I, the major apolipoprotein component of HDL, promotes ABCA1-mediated cholesterol and phospholipid efflux, probably by directly binding to ABCA1. ABCA1 gene expression is markedly increased in cholesterol loaded cells as a result of activation of LXR/RXR. ABCA1 protein turnover is rapid. ABCA1 contains a PEST--proline (P), glutamate (E), serine (S), and threonine (T)--sequence in the intracellular segment that mediates ABCA1 degradation by a thiol protease, calpain. ApoA-I and apoE stabilize ABCA1 in a novel mode of regulation by decreasing PEST sequence-mediated calpain proteolysis. ABCA1-mediated cholesterol and phospholipid efflux are distinctly regulated and affected by the activity of other gene products. Stearyol CoA desaturase decreases ABCA1-mediated cholesterol efflux but not phospholipid efflux, likely by decreasing the cholesterol pool available to ABCA1. This and other evidence suggest that ABCA1 promotes cholesterol and phospholipid efflux, probably by directly transporting both lipids as substrates. PMID- 12738682 TI - Identification and quantification of diadenosine polyphosphate concentrations in human plasma. AB - OBJECTIVE: Diadenosine polyphosphates have been demonstrated to be involved in the control of vascular tone as well as the growth of vascular smooth muscle cells and hence, possibly, in atherogenesis. In this study we investigated the question of whether diadenosine polyphosphates are present in human plasma and whether a potential source can be identified that may release diadenosine polyphosphates into the circulation. METHODS AND RESULTS: Plasma diadenosine polyphosphates (ApnA with n=3 to 6) were purified to homogeneity by affinity-, anion exchange-, and reversed phase-chromatography from deproteinized human plasma. Analysis of the homogeneous fractions with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) revealed molecular masses ([M+H]+) of 757, 837, 917, and 997 d. Comparison of the postsource decay matrix assisted laser desorption/ionization mass spectrometry mass spectra of these fractions with those of authentic diadenosine polyphosphates revealed that these isolated substances were identical to Ap3A, Ap4A, Ap5A, and Ap6A. Enzymatic analysis showed an interconnection of the phosphate groups with the adenosines in the 5'-positions of the ribose moieties. The mean total plasma diadenosine polyphosphate concentrations (micromol/L; mean +/- SEM) in cubital veins of normotensive subjects amounted to 0.89+/-0.59 for Ap3A, 0.72+/-0.72 for Ap4A, 0.33+/-0.24 for Ap5A, and 0.18+/-0.18 for Ap6A. Cubital venous plasma diadenosine polyphosphate concentrations from normotensives did not differ significantly from those in the hypertensive patients studied. There was no significant difference between arterial and venous diadenosine polyphosphate plasma concentrations in 5 hemodialysis patients, making a significant degradation by capillary endothelial cells unlikely. Free plasma diadenosine polyphosphate concentrations are considerably lower than total plasma concentrations because approximately 95% of the plasma diadenosine polyphosphates were bound to proteins. There were no significant differences in the diadenosine polyphosphate plasma concentrations depending on the method of blood sampling and anticoagulation, suggesting that platelet aggregation does not artificially contribute to plasma diadenosine polyphosphate levels in significant amounts. The ApnA (with n=3 to 6) total plasma concentrations in adrenal veins were significantly higher than the plasma concentrations in both infrarenal and suprarenal vena cava: adrenal veins: Ap3A, 4.05+/-1.63; Ap4A, 6.18+/-2.08; Ap5A, 0.53+/-0.28; Ap6A, 0.59+/-0.31; infrarenal vena cava: Ap3A, 1.25+/-0.66; Ap4A, 0.91+/-0.54; Ap5A, 0.25+/-0.12; Ap6A, 0.11+/ 0.06; suprarenal vena cava: Ap3A, 1.40+/-0.91; Ap4A, 1.84+/-1.20; Ap5A, 0.33+/ 0.13; Ap6A, 0.11+/-0.07 (micromol/L; mean +/- SEM; each P<0.05 (concentration of adrenal veins versus infrarenal or suprarenal veins, respectively). CONCLUSIONS: The presence of diadenosine polyphosphates in physiologically relevant concentrations in human plasma was demonstrated. Because in adrenal venous plasma significantly higher diadenosine polyphosphate concentrations were measured than in plasma from the infrarenal and suprarenal vena cava, it can be assumed that, beside platelets, the adrenal medulla may be a source of plasma diadenosine polyphosphates in humans. PMID- 12738683 TI - Development of an animal model for spontaneous myocardial infarction (WHHLMI rabbit). AB - OBJECTIVE: Coronary heart disease is the most common cause of death in developed countries. However, there are no suitable animal models that mimic spontaneous myocardial infarction in humans. In this study, we attempted to obtain a rabbit strain with spontaneous myocardial infarction by selective breeding of coronary atherosclerosis-prone Watanabe heritable hyperlipidemic (WHHL) rabbits, designated as WHHLMI rabbits. METHODS AND RESULTS: WHHLMI rabbits were characterized by the high incidence of fatal myocardial infarction at ages 11 to 35 months, being increased from 23% to 97% after the selective breeding. The ECG on WHHLMI rabbits showed a typical feature of myocardial infarction. Histological examination of hearts from suddenly deceased WHHLMI rabbits revealed old myocardial infarction accompanied by fresh myocardial lesions. The culprit coronary arteries exhibited severe atheromatous plaques (>90% lumen area stenosis), suggesting that coronary atherosclerosis is responsible for myocardial infarction observed in WHHLMI rabbits. In addition, the coronary plaques showed vulnerable features including macrophage-rich thin cap and large necrotic core. CONCLUSIONS: To the best of our knowledge, this is the first report of spontaneous myocardial infarction in rabbits, and it is suggested that this WHHLMI rabbit strain will be a useful animal model to study human myocardial infarction. PMID- 12738685 TI - Lipopolysaccharide induces preprotachykinin gene expression. AB - This study was performed to test whether biosynthesis of tachykinins plays a pivotal role in lipopolysaccharide (LPS)-induced airway alteration by analyzing preprotachykinin-I (PPT-I, a precursor of tachykinins) gene expression. Brown Norway rats (11-12 wk old) were divided into four groups: control; LPS; dimethylthiourea (DMTU, an effective hydroxyl radical scavenger); and DMTU+LPS. Each animal in the control group received saline treatment. Forty-nine animals in the LPS group were further divided into seven subgroups to test effects of doses and length of the LPS treatment. Total RNA extracted from nodose ganglia and lungs was used to assay relative amount of PPT-I mRNA using the real-time quantitative reverse transcriptase-polymerase chain reaction. In addition, LPS induced alterations in airway responses to bronchial constrictors, neutral endopeptidase (NEP) gene expression, leukocyte counts, and SP and calcitonin gene related peptide (CGRP) levels were determined. LPS (4 mg/kg, intraperitoneal) raised significantly PPT-I mRNA level after 4 h in nodose ganglia and 12 h in the lung, and this elevation sustained for 5 d. Also, LPS caused significant increases in NEP mRNA, SP and CGRP levels, airway reactivity to capsaicin and SP, and neutrophil counts, but a significant decrease in macrophage count. Our data support that LPS-induced bronchial hyperreactivity to capsaicin is related closely to the upregulation of tachykinin gene expression, but not the upregulation of NEP. PMID- 12738684 TI - Molecular basis of platelet granule secretion. AB - The energy-dependent release of granule contents from activated platelets is a well-established component of normal hemostasis and thrombosis. A role for membrane fusion in this process has been presumed for decades, but only recently have the mechanisms of platelet membrane fusion been investigated at the molecular level. Such studies have demonstrated that platelet membrane fusion is controlled by lipid components of the membrane bilayer, by transmembrane proteins termed SNARE proteins, and by chaperone proteins that interact with SNARE proteins. This core membrane fusion machinery is controlled by activation dependent changes in cytoskeletal organization, intracellular calcium levels, kinase activity, and intracellular protease activity. Through these mechanisms, interactions of ligands with their cognate cell-surface receptors are transmitted to the membrane fusion machinery to facilitate membrane fusion and secretion of granule contents from platelets. PMID- 12738686 TI - Low molecular weight hyaluronan from stretched lung enhances interleukin-8 expression. AB - Mechanical ventilation has been shown to cause ventilator-induced lung injury (VILI), probably by overdistending or stretching the lung. Hyaluronan (HA), a component of the extracellular matrix, in low molecular weight (LMW) forms has been shown to induce cytokine production. LMW HA is produced by hyaluronan synthase 3 (HAS 3). We found that HAS 3 mRNA expression was upregulated and that LMW HA accumulated in an animal model of VILI. We hypothesized that stretch induced LMW HA production that causes cytokine release in VILI was dependent on HAS 3 mRNA expression. We explored this hypothesis with in vitro lung cell stretch. Cell stretch induced HAS 3 mRNA expression and LMW HA in fibroblasts. Nonspecific inhibitors of HAS 3 (cyclohexamide and dexamethasone), a nonspecific inhibitor of protein tyrosine kinases (genistein), and a janus kinase 2 inhibitor (AG490) blocked stretch-induced HAS 3 expression and synthesis of LMW HA. Stretch induced LMW HA from fibroblasts caused a significant dose-dependent increase in interleukin-8 production both in static and stretched epithelial cells. These results indicated that de novo synthesis of LMW HA was induced in lung fibroblasts by stretch via tyrosine kinase signaling pathways, and may play a role in augmenting induction of proinflammatory cytokines in VILI. PMID- 12738687 TI - Prostaglandin E2 inhibits fibroblast to myofibroblast transition via E. prostanoid receptor 2 signaling and cyclic adenosine monophosphate elevation. AB - Myofibroblasts, the hallmark of fibrotic disease, contribute to the pathology of fibrosis by secreting large amounts of extracellular matrix and contributing to alveolar contraction. Myofibroblasts are characterized by the expression of alpha smooth muscle actin (alpha-SMA), a contractile protein normally associated with smooth muscle cells. Transforming growth factor-beta1 (TGF-beta1) is a well characterized profibrotic cytokine that induces myofibroblast transformation both in vitro and in vivo. We report here that the lipid mediator prostaglandin E2 (PGE2) inhibits TGF-beta1-induced expression of alpha-SMA in primary fetal and adult lung fibroblasts. This inhibition of alpha-SMA expression is associated with a reduction in the expression of collagen I. Inhibitory actions of PGE2 are mediated via E prostanoid receptor 2 (EP2) signaling, but not by EP3 signaling, and increases in cyclic adenosine monophosphate production. The inhibitory effects of PGE2 on TGF-beta1-induced alpha-SMA expression are mimicked by an EP2 selective agonist, butaprost, and by forskolin-induced direct activation of adenyl cyclase. An EP2 antagonist blocks the inhibitory effects of PGE2, and an EP3 agonist does not inhibit TGF-beta1-mediated increases in alpha-SMA expression. Our results demonstrate that PGE2 inhibits transition of fibroblasts to myofibroblasts by an EP2 receptor-activated pathway. Augmenting this pathway may serve as a potent antifibrotic therapeutic strategy. PMID- 12738688 TI - Serine protease inhibitors modulate smoke-induced chemokine release from human lung fibroblasts. AB - Smoking is associated with lung inflammation and a protease-antiprotease imbalance. We previously reported that cigarette smoke extract (CSE) stimulates human lung fibroblasts to release chemotactic cytokines. We hypothesized that serine protease inhibitors might modulate lung fibroblast release of chemotactic cytokines in response to CSE. To test this hypothesis, serine protease inhibitors (FK706, alpha1-antitrypsin, methoxysuccinyl-Ala-Ala-Pro-Val chloromethyl ketone, or Nalpha-p-tosyl-L-lysine chloromethyl ketone) were evaluated for their capacity to attenuate the release of neutrophil chemotactic activity (NCA) and monocyte chemotactic activity (MCA) from human fetal lung fibroblasts by the blind-well chemotactic chamber. Metalloproteinases and cysteine proteinases were not examined in this study. Similarly, the release and gene expression of chemokines and nuclear factor-kappaB (NF-kappaB) activation were measured by means of enzyme linked immunosorbent assay and reverse transcriptase-polymerase chain reaction. Release of NCA, MCA, chemotactic chemokines including interleukin-8, granulocyte colony-stimulating factor, monocyte chemoattractant protein-1, and granulocyte macrophage colony-stimulating factor, and the expression of interleukin-8 and monocyte chemoattractant protein-1 mRNA were attenuated by FK706. Furthermore, FK706 suppressed NF-kappaB activation. These data suggest that serine protease inhibitors attenuate the CSE-induced release of NCA and MCA from human fetal lung fibroblasts and that the inhibitory action of antiproteases might depend on NF kappaB signaling pathway. PMID- 12738689 TI - The frequency and nature of medical error in primary care: understanding the diversity across studies. AB - BACKGROUND: The identification and reduction of medical error has become a major priority for all health care providers, including primary care. Understanding the frequency and nature of medical error in primary care is a first step in developing a policy to reduce harm and improve patient safety. There has been scant research into this area. OBJECTIVES: This review had two objectives; first, to identify the frequency and nature of error in primary care, and, secondly, to consider the possible causes for the diversity in the stated rates and nature of error in primary care. METHODS: Literature searches of English language studies identified in the National Patient Safety Foundation bibliography database, in Medline and in Embase were carried out. Studies that were relevant to the purpose of the study were included. Additional information was obtained from a specialist medico-legal database. RESULTS: Studies identified that medical error occurs between five and 80 times per 100000 consultations, mainly related to the processes involved in diagnosis and treatment. Prescribing and prescription errors have been identified to occur in up to 11% of all prescriptions, mainly related to errors in dose. There are a wide variety of definitions and methods used to identify the frequency and nature of medical error. Incident reporting, systematic identification and medico-legal databases reveal differing aspects, and there are additional perspectives obtained from GPs, primary health care workers and patients. CONCLUSION: An understanding of the true frequency and nature of medical error is complicated by the different definitions and methods used in the studies. Further research is warranted to understand the complex nature and causes of such errors that occur in primary care so that appropriate policy decisions to improve patient safety can be made. PMID- 12738690 TI - Error and safety in primary care: no clear boundaries. AB - This paper examines the notions of adverse events, error, critical incidents and safety from the specific viewpoint of primary care. We conclude that each term can be defined, but existing work which we reviewed uses many of the terms interchangeably. We recognise that trying to access medical error objectively within primary care can be problematic. Regardless of definitions, reflection on critical incidents, adverse events or other notable events is important, but requires time and resources to be conducted effectively. PMID- 12738691 TI - "Fire away": the opening sequence in general practice consultations. AB - BACKGROUND: Proponents of recent models of the doctor-patient relationship, such as concordance and shared decision making, have emphasized mutuality rather than paternalism or consumerism. However, little attention has been paid so far to the ways in which this might actually be achieved. OBJECTIVES: The aims of this study were to establish whether there are any rules governing the opening sequence in general practice consultations, and to analyse the ways in which the observing or breaking of such rules contributes to the development of mutuality between patients and GPs. METHODS: The paper is based on a qualitative study of 62 patients consulting 20 GPs in 20 practices in the Midlands and Southeast of England. Consultations were audio recorded and transcribed; patients were interviewed before and after each consultation, and doctors were interviewed afterwards. Data were analysed using the sociological method of Conversation Analysis. The outcomes were participants' own understandings as demonstrated in their speech. RESULTS: A selection rule was identified whereby doctors choose between the questions "How are you?" and "What can I do for you?" to elicit patients' concerns. Deviations from this selection rule may be either repairable or strategic. Repairable deviance is based on misunderstanding between participants, and is resolved interactionally, usually by patients. Strategic deviance is the attempt by doctors to emphasize or de-emphasize certain aspects of their relationships with particular patients. Deviations from the rule which are not repaired lead to misalignment between participants. CONCLUSION: In relation to concordance, or shared decision making more generally, this analysis demonstrates that alignment or misalignment between participants will occur before any discussion about treatment options occurs. In cases of misalignment, concordance will be much harder to achieve. Mutuality is an achievement of both patients and doctors, and requires the active participation of patients. PMID- 12738692 TI - What's in a name? An experimental study of patients' views of the impact and function of a diagnosis. AB - OBJECTIVE: The aim of the present study was to examine patients' views about the relative impact and function of lay and medical diagnoses for stomach and throat problems. METHODS: A questionnaire survey was carried out among 900 consecutive patients attending nine general practices across England. A total of 740 questionnaires were completed (response rate: 82.2%). Each participant rated a series of statements describing the impact upon the patient and the function for the doctor following both a stomach and a throat problem case scenario involving either a lay (stomach upset/sore throat) or medical (gastroenteritis/tonsillitis) label. RESULTS: The results showed consistent differences between the lay and medical labels for both stomach and throat problems in terms of their impact upon the patient and their function for the doctor. In particular, the medical labels were rated as beneficial for the patient in terms of validating the sick role and improving their confidence in the doctor. In contrast, the lay labels resulted in a greater sense of ownership of the problem which could be associated with unwanted responsibility and blame. In addition, the medical labels were seen to provide the doctor with a greater sense of professionalism, as giving them a clearer role in the consultation and to imply less blame on the part of the patient. 'Stomach upset' was also seen as a more pragmatic label than 'gastroenteritis'. CONCLUSION: Although much current prescriptive literature in general practice advocates the use of lay language in the consultation as a means to promote better doctor-patient partnerships, the issue of diagnosis is more complex than this. Patients attribute greater benefits to the use of medical labels for themselves and state that such medical labels are of greater benefit to the doctor. PMID- 12738693 TI - GPs facing reluctant and demanding patients: analysing ethical justifications. AB - BACKGROUND: Several studies have explored the physicians' preferred actions when facing a reluctant or a demanding patient, but only a few studies have explored the physicians' justifying reasons. OBJECTIVE: The aim of this study was to assess how GPs would act and how they would justify their choice. METHOD: A postal questionnaire with questions about preferred actions and justifying reasons was sent to a random sample of GPs in Slovenia (n = 160) and Sweden (n = 200) using four vignettes: (i). a healthy patient reluctant to quit smoking; (ii). a healthy patient demanding an X-ray; (iii). a pulmonary cancer patient reluctant to quit smoking; and (iv). a pulmonary cancer patient demanding immunotherapy. RESULTS: The majority of GPs would bring up the question about smoking with the patients reluctant to quit. They justified their choice by referring to promotion of medical benefit and to protection from harm. Swedish GPs were less inclined to bring up smoking than were their Slovenian colleagues. Those who would not bring up the question referred to respect for self determination and an enhanced relationship as their justifying reasons. With reference to the demanding patients, a minority of GPs would grant the healthy patient's request for an X-ray that was not medically motivated. The answers were similar with respect to the seriously ill patient requesting non-medically motivated immunotherapy. Slovenian GPs were much more inclined to grant the request than were their Swedish colleagues. Enhancing the relationship and respect for self-determination were the most important reasons for granting the demands. When the demands were denied, the GPs mostly referred to promotion of fair distribution of resources. CONCLUSION: Many of the GPs considered their patients' right to self-determination less important than other values, e.g. the obligation to promote medical benefit, to protect from harm, to distribute public resources fairly and to enhance the patient-physician relationship. PMID- 12738694 TI - GPs' perspectives on managing time in consultations with patients suffering from depression: a qualitative study. AB - BACKGROUND: Although there is widespread concern that general practice consultations are too short for doctors to provide a high quality of care for patients, the relationship between the length and outcome of these consultations remains unclear. Research to date has neglected the subjective experience of consultation time of both patients and GPs. OBJECTIVES: Our aim was to investigate GP perspectives on consultation time and the management of depression in general practice. METHOD: A qualitative interview-based study was carried out of 19 GPs from eight West Midlands general practices. RESULTS: The GPs in this study acknowledged the pressure of work and resource constraints in general practice. However, they did not feel these prevented them from providing good support and treatment for depression. They were confident in the effectiveness of antidepressants and their own skills in providing counselling support, and were able to utilize time flexibly in responding to patients' variable needs. Depression was viewed as a relatively straightforward problem that usually could be managed within the resources available to general practice. CONCLUSION: The doctors generally did not experience time to be a limiting factor in providing care for patients with depression. This is in contrast to the more acute sense of time pressure commonly reported by patients which they felt undermined their capacity to benefit from the consultation. GPs need to be more aware of patient anxieties about time, and to devise effective means of raising patients' sense of time entitlement in general practice consultations. PMID- 12738695 TI - Factors influencing inquiry about patients' alcohol consumption by primary health care physicians: qualitative semi-structured interview study. AB - BACKGROUND: Early recognition of and intervention in risky alcohol consumption has been shown to be an effective way to reduce the harm. However, primary care physicians are still not screening for and intervening sufficiently in their patients' alcohol misuse. OBJECTIVE: The purpose of this study was to explore factors having an effect on primary health care physicians inquiring about patients' alcohol consumption. METHOD: A qualitative study of primary care physicians' experiences and views based on tape recorded semi-structured interviews was carried out on all physicians (n = 35) working at four health centres in Eastern Finland. RESULTS: Seven main categories were identified that either prevent or promote discussion about alcohol consumption: the sensitive nature of alcohol drinking; the reason for consultation; awareness of a patient's alcohol problem; patient factors; availability of intervention tools; expectations of effectiveness of interventions; and lack of time. CONCLUSIONS: There still exist many barriers to initiating discussions about alcohol in the consultation room. Changing the frame of reference of the concept of alcohol drinking from an addictive disease to a general lifestyle risk factor could overcome many of these barriers. PMID- 12738696 TI - Doctor-patient communication: a comparison of the USA and Japan. AB - BACKGROUND: Little is known about the differences and similarities between doctor patient communication patterns in different cultures. OBJECTIVES: The aim of this study was to examine communication patterns of doctor-patient consultations in two different cultures, namely the USA and Japan, and to elucidate linguistic differences and similarities in communication. METHODS: This cross-sectional study used quantitative discourse analysis from linguistics to compare 40 doctor patient consultations: 20 out-patient consultations of five physicians in the USA and 20 out-patient consultations of four physicians in Japan. The main outcomes measured were time spent in each phase of the encounter, number of categorized speech acts, distribution of question types and frequencies of back-channel responses and interruptions. RESULTS: The average length of doctor-patient encounters was 668.7 s in the USA and 505 s in Japan. US physicians spent relatively more time on treatment and follow-up talk (31%) and social talk (12%), whereas the Japanese had longer physical examinations (28%) and diagnosis or consideration talk (15%). Japanese doctor-patient conversations included more silence (30%) than those in the USA (8.2%). The doctor-patient ratios of total speech acts were similar (USA 55% versus 45%; Japan 59% versus 41%). Physicians in both countries controlled communication during encounters by asking more questions than the patients (75% in the USA; 78% in Japan). The Japanese physicians and patients used back-channel responses and interruptions more often than those in the USA. CONCLUSIONS: While doctor-patient communication differed between the USA and Japan in the proportion of time spent in each phase of the encounter, length of pauses and the use of back-channel responses and interruptions, physician versus patient ratios of questions and other speech acts were similar. The variations may reflect cultural differences, whereas the similarities may reflect professional specificity stemming from the shared needs to fill the information gap between physician and patient. Adequate awareness of these differences and similarities could be used to educate clinicians about the best approaches to patients from particular cultural backgrounds. PMID- 12738697 TI - GPs' satisfaction with the doctor-patient encounter: findings from a community based survey. AB - OBJECTIVE: The aim of this study was to explore the characteristics of the doctor patient relationship from the GP's point of view. METHODS: We performed a cross sectional 1-day study in family practice. Thirty-three GPs volunteered to fill in a questionnaire at the end of each of 20 consecutive consultations on an index day. Six hundred and sixty-one patients (out of 665) participated in the study. Descriptive frequencies of GPs' judgements about personal experiences during the consultations, and predictors of GP's global satisfaction score on patient encounters were analysed. RESULTS: The mean age of the 33 GPs was 44.7 +/- 3.6 years. Professional skills (62% of the GPs had no doubts on diagnosis, therapy or prognosis) and the quality of the human/interpersonal interaction were major determinants of GPs' satisfaction in the patient-doctor relationship. Doctors felt professionally esteemed by 90% of their patients, and the median value of their global satisfaction score (matching the expectations from an 'ideal patient' to that experienced when meeting the real one) was very high (median 8, range 1-10). Nevertheless, GPs did not know if they were satisfied with the actual encounter with the patient in about one-third of the consultations. CONCLUSIONS: Professional skills and quality of the human/interpersonal interactions are major determinants of GPs' satisfaction in their professional activities. PMID- 12738698 TI - Osteoporosis: what are the implications of DEXA scanning 'high risk' women in primary care? AB - BACKGROUND: Current recommended practice for the use of dual X-ray absorptiometry (DEXA) scans in screening for osteoporosis is to concentrate on women at 'high risk'. OBJECTIVE: We have applied such a screening strategy, in a general practice setting, to estimate the number of women requiring scans. METHODS: A two phase survey was carried out: (i). postal screen of clinical indicators for low bone mineral density (BMD) to define women at 'high risk'; and (ii). DEXA scanning of the sample at 'high risk' set in two general practices in North Staffordshire. Computerized general practice records were used to define a purposive sample of 1001 women, to receive the screening tool, consisting of three equal size groups (i). those with an early hysterectomy; (ii). those receiving oral corticosteroids on repeat prescription; and (iii). those on the practice cervical smear register. A random sample of women defined at 'high risk' by the screening tool were invited to have a DEXA scan. The main outcome of interest was the presence of low BMD as measured by a DEXA scanner. RESULTS: Sixty-five out of 95 women invited (68%) agreed to undergo a DEXA scan: median age = 52 years (interquartile range 44-64 years). Twenty-nine of these 65 women (45%) were classified with low BMD (WHO criteria): 90% had densities below their age-matched mean. Extrapolating from the observed findings to the main study practice (n = 9000 total population), we estimate that 162 women would be defined at 'high risk', and, if all were offered a scan, 105 would comply and 56 would be defined with low BMD. CONCLUSIONS: Using this approach, we estimate the unmet need, in women, for DEXA scans to be 180 per 10000 total practice population. Allowing for scan uptake, this would define approximately 60 women per 10000 total practice population with low BMD. The application of this screening strategy has identified a group of women who might benefit from treatment or prophylaxis for osteoporosis. PMID- 12738699 TI - GP and patient predictors of PSA screening in Australian general practice. AB - OBJECTIVE: We determined GP and patient variables associated first with men's prior uptake of prostate-specific antigen (PSA) screening and, subsequently, its initiation during an 'index consultation' in Australian general practice. METHODS: From the practices of 60 GPs, we recruited a sample of 423 male patients aged 40-70 years. In a waiting room questionnaire completed before their 'index consultation' (retrospective component), men reported their previous PSA screening status. We obtained demographic and clinical data, including the presence of lower urinary tract symptoms (LUTS). Men also were mailed a questionnaire 2 days after their 'index consultation' to ascertain whether the GP had discussed PSA screening (prospective component) for prostate cancer and other behaviours. GPs themselves completed questionnaires eliciting demographic and practice characteristics as well as their propensity to screen and understanding of the evidence about PSA testing. GP and patient study variables were modelled simultaneously in analyses. RESULTS: Of those 348 men consulting with their regular GP, 80 (23.0%) reported previously having had a PSA screening test. Men were significantly and independently more likely ever to have had PSA screening if their regular GP reported a propensity to initiate screening [adjusted odds ratio (AOR) = 2.27, 95% confidence interval (CI) 1.23-4.20; P = 0.009]. GP age also was independently associated with men's PSA screening status [chi-squared (3) P < 0.0001] as was men's age and severity of LUTS (AOR = 2.38, 95% CI 1.58 3.57, P < 0.0001 and AOR = 1.79, 95% CI 1.00-3.19, P = 0.004, respectively). Current smokers were less likely ever to have had a PSA screening test (AOR = 0.34, 95% CI 0.16-0.69; P = 0.003). Discussion of PSA screening in their 'index consultation' was recalled independently more often by older men (AOR = 1.46, 95% CI 1.00-2.13; P = 0.04), those with moderate/severe LUTS (AOR = 1.94, 1.07-3.49; P = 0.04), those whose GP had performed or discussed a cholesterol test (AOR = 2.26, 95% CI 1.03-4.92; P = 0.04) and those whose GP had postgraduate training in family medicine (AOR = 3.13, 95% CI 1.23-8.00; P = 0.02). CONCLUSION: In the absence as yet of compelling evidence that PSA screening will prolong life or enhance its quality, our findings identify GP and patient factors that could be targeted to modify PSA screening. PMID- 12738700 TI - Organizing palliative care for rural populations: a systematic review of the evidence. AB - BACKGROUND: Palliative care services have developed mostly in urban areas. Rural areas typically are characterized by the lack of well-organized services, with primary care professionals, specifically GPs and community nurses, having to undertake most of the palliative care. Little is known, however, either of their views or of how best to organize palliative care in rural areas. OBJECTIVE: The aim of this study was to conduct a systematic literature review of studies that have examined the organization of rural palliative care and the views of professionals in rural areas. METHOD: Six electronic databases were searched for published studies between 1991 and 2001. Articles had to match against (i). MeSH or keyword terms relating to palliative, terminal or end of life care; and (ii). MeSH or keyword terms relating to rural. A data extraction framework was designed and used systematically by two reviewers to consider research question and method, sample characteristics, selection and size, study quality, summary results and implications. RESULTS: Twenty-six studies were identified. These were mostly questionnaire surveys and reports, and three qualitative studies. No randomized controlled studies or cohort studies were identified. Education and strategic issues were dominant research questions. Both the sample numbers and response rates in the surveys were variable. The qualitative studies had methodological strengths and elicited important views from nurses, carers and families. GPs were, however, unrepresented. Whilst the role of primary care emerged as an important theme, primary care professionals reported difficulties in obtaining education and training. There were also reported problems in symptom control and in the management of emotional issues such as bereavement counselling. Difficulties were also described in accessing specialist services such as hospices, and families were reported as having problems in accessing information. Developments in information technology such as telemedicine were seen as possible solutions to some of the problems. CONCLUSIONS: There is little published work on this topic. Most of the work identifies problems in the delivery of palliative care in rural areas. Whilst primary care professionals are seen as having a key role, there is a need to discover both their views and their needs in this field. PMID- 12738701 TI - A systematic review of the effect of different models of after-hours primary medical care services on clinical outcome, medical workload, and patient and GP satisfaction. AB - BACKGROUND: The organization of after-hours primary medical care services is changing in many countries. Increasing demand, economic considerations and changes in doctors' attitudes are fueling these changes. Information for policy makers in this field is needed. However, a comprehensive review of the international literature that compares the effects of one model of after-hours care with another is lacking. OBJECTIVE: The aim of this study was to carry out a systematic review of the international literature to determine what evidence exists about the effect of different models of out-of-hours primary medical care service on outcome. METHODS: Original studies and systematic reviews written since 1976 on the subject of 'after-hours primary medical care services' were identified. Databases searched were Medline/Premedline, CINAHL, HealthSTAR, Current Contents, Cochrane Reviews, DARE, EBM Reviews and EconLit. For each paper where the optimal design would have been an interventional study, the 'level' of evidence was assessed as described in the National Health and Medical Research Council Handbook. 'Comparative' studies (levels I, II, III and IV pre-/post-test studies) were included in this review. RESULTS: Six main models of after-hours primary care services (not mutually exclusive) were identified: practice-based services, deputizing services, emergency departments, co-operatives, primary care centres, and telephone triage and advice services. Outcomes were divided into the following categories: clinical outcomes, medical workload, and patient and GP satisfaction. The results indicate that the introduction of a telephone triage and advice service for after-hours primary medical care may reduce the immediate medical workload. Deputizing services increase immediate medical workload because of the low use of telephone advice and the high home visiting rate. Co operatives, which use telephone triage and primary care centres and have a low home visiting rate, reduce immediate medical workload. There is little evidence on the effect of different service models on subsequent medical workload apart from the finding that GPs working in emergency departments may reduce the subsequent medical workload. There was very little evidence about the advantages of one service model compared with another in relation to clinical outcome. Studies consistently showed patient dissatisfaction with telephone consultations. CONCLUSIONS: The rapid growth in telephone triage and advice services appears to have the advantage of reducing immediate medical workload through the substitution of telephone consultations for in-person consultations, and this has the potential to reduce costs. However, this has to be balanced with the finding of reduced patient satisfaction when in-person consultations are replaced by telephone consultations. These findings should be borne in mind by policy makers deciding on the shape of future services. PMID- 12738702 TI - A framework for clinical general practice and for research and teaching in the discipline. AB - This paper uses three typical case stories from general practice to demonstrate that a GP simultaneously considers four dimensions when making a diagnosis and planning subsequent treatment of a patient in the consultation: (i). a biomedical dimension; (ii). a culture and context dimension; (iii). a medico-psychological dimension; and (iv). a network and social dimension. By taking this diagnostic and therapeutic approach, the GP adds value to the total performance of the health care system. It is demonstrated that a GP needs theoretical, research based knowledge and skills within all four dimensions, and that it is necessary for a GP to work together with both medical and non-medical disciplines when defining the research and teaching agenda. It is stressed that consultation and communication skills are important tools for any doctor, and the value of continuity of care is discussed. Finally, the implications of the diagnostic approach with respect to planning research and teaching programmes are discussed, and the need for a better balance is stressed. PMID- 12738703 TI - An approach for integrating complementary-alternative medicine into primary care. AB - BACKGROUND: Despite family practitioners frequently being requested to assist their patients with advice on or referrals to complementary-alternative medicine (CAM), there is an absence both of evidence about the efficacy of nearly all specific treatments or modalities and of guidelines to assist with the integration of conventional and CAM therapies. OBJECTIVE: The aim of this article is to suggest a comprehensive and rational, best-evidence strategy for integrating CAM by primary care practitioners into primary care, within the context of the limitations of the current knowledge base and the local milieu. METHODS: The suggested approach was developed by a combination of literature review, key informant interviews, focus groups, educational presentations for family practice residents and practitioners, and field testing. An iterative model was utilized whereby more refined drafts of the suggested approach were subjected to later discussants and groups, as well as further field testing. Drafts of the strategy were utilized in consultations of patients requesting advice on alternative medicine in a primary care setting and in a CAM clinic. RESULTS: Both family physicians and CAM practitioners provided useful comments and recommendations throughout the process. These can be categorized in terms of knowledge, attitudes and skills. Our strategy suggests that patients requesting advice on the use and integration of CAM modalities as part of their health care should be evaluated initially by their primary care physician. The physician's responsibilities are to evaluate the appropriateness of that use, and to maintain contact, monitoring outcomes. Advice on referrals should be based on the safety of the method in question, current knowledge on indications and contraindications of that modality, and familiarity and an open dialogue with the specific therapist. CONCLUSIONS: Given patients' demands and utilization of CAM therapies, despite the lack of evidence, there is an increasing need to address how CAM therapies can be integrated into conventional medical systems. These suggestions should respond to patient's expectations and needs, but at the same time maintain accepted standards of medical and scientific principles of practice. PMID- 12738704 TI - Process evaluation of a cluster randomized trial of tailored interventions to implement guidelines in primary care--why is it so hard to change practice? AB - BACKGROUND: A cluster randomized trial of tailored interventions to support the implementation of guidelines for sore throat and urinary tract infection found little or no change in the main outcomes, which were antibiotic prescriptions, use of laboratory tests and use of telephone consultations. There was great variation between the practices in the change in these outcomes. OBJECTIVES: Our aim was to evaluate how the interventions were received and to understand why practices did or did not change. METHODS: The trial was conducted in general practices in Norway. Data for this process evaluation were collected from the 120 practices that completed the trial. Multiple methods were used: observations, semi-structured telephone interviews, a postal survey and data extracted from electronic medical records. We investigated factors that might explain a lack of change, including: agreement with the guidelines; communication within each practice; degree of participation in the project; taking time to discuss the guidelines and their implementation; use of the components of the interventions; and routines for telephone consultations. Possible explanatory factors were explored in relation to variation in change and the overall extent of change in rates of use of antibiotics, laboratory tests and telephone consultations. RESULTS: Sixty-three per cent of practices agreed with the guidelines. Only 35% reported having regular meetings, and 33% discussed the project before its start, although 75% reported agreement about participating within the practice. Only 33% reported meeting to discuss the guidelines. Use of the components of the interventions ranged from 11% for the increased fee for telephone consultations to 48% for the computerized decision support. Forty-four per cent reported problems with telephone routines. No single factor explained the observed variation in the extent of change across practices. CONCLUSIONS: Inadequate time, resources and support were the most salient factors that might explain a lack of change. Problems with internal communication and telephone routines were important contributing factors in many practices. PMID- 12738705 TI - Developing clinically relevant and reproducible symptom-defined populations for cancer diagnostic research in general practice using a community survey. AB - BACKGROUND: In developing cancer diagnostic studies, there is a particular need for the population of patients with symptoms of possible oncological significance who consult the GP in 'real life' to be comparable with the group of individuals with symptoms constructed as part of a research project. OBJECTIVES: The objective of this study was to seek to assess whether a community-based symptom survey can be harnessed in order to produce clinically relevant and reproducible populations within which studies of more detailed indicants could be undertaken. METHOD: A total of 3629 patients registered with a general practice at Winterton, UK, were sent a questionnaire enquiring about 10 symptoms of possible oncological significance together with their consultation intention in relation to these symptoms. Up to 1 month later, an identical questionnaire was applied to all patients reporting at least one symptom, and more detailed information was obtained by research nurses on each symptom. RESULTS: The overall response rate was 64.4%, and 850 patients reported one or more symptoms. For the majority of symptom reports, there was moderate to substantial agreement between the two applications of the questionnaire. The question on blood in the motions/toilet pan or on the toilet paper demonstrated almost perfect agreement. Slight agreement was found for abdominal pain for longer than 4 weeks and for black/tarry motions. In relation to the reliability of the patient consultation intention, there was substantial/moderate agreement for actions related to the majority of symptoms. For all symptoms, there was also a greater level of agreement for past activity than future intent. CONCLUSION: The results of the study provide some support for a community survey as a mechanism to develop 'clinically relevant' populations for the iatrotropic symptoms rectal bleeding or indigestion/heartburn within which studies of more detailed indicants could be undertaken. There is also consistency with the work of others in relation to the numbers and characteristics of patients within the 'clinically relevant' population. PMID- 12738706 TI - Selections from current literature: benzodiazepines revisited. PMID- 12738707 TI - Coronary heart disease: acceptance of guideline recommendations irrelevant in Germany? PMID- 12738708 TI - Evaluation of the whole prostaglandin biosynthetic pathway in lung cancer. PMID- 12738709 TI - Clinical trial design and end points for epidermal growth factor receptor targeted therapies: implications for drug development and practice. PMID- 12738710 TI - Tumors and their microenvironments: tilling the soil. Commentary re: A. M. Scott et al., A Phase I dose-escalation study of sibrotuzumab in patients with advanced or metastatic fibroblast activation protein-positive cancer. Clin. Cancer Res., 9: 1639-1647, 2003. PMID- 12738711 TI - Reactivation of tumor suppressor genes by the cardiovascular drugs hydralazine and procainamide and their potential use in cancer therapy. AB - PURPOSE: The purpose of this study is to evaluate the demethylating and tumor suppressor-reactivating activity of hydralazine and procainamide. EXPERIMENTAL DESIGN: MDA-231, MCF-7, and T24 cell lines were treated for 5 days with 10 micro M hydralazine or 10 micro M procainamide. 5-aza-deoxycytidine at 0.75 micro M was used as positive control. BALB/c nu/nu mice xenografted with MDA-231 cells were treated with these drugs for 7 days by i.p. route. Methylation was assessed by PCR after digestion with methylation-sensitive enzymes for the ER gene and with methylation-specific PCR for retinoic acid receptor (RAR)beta and p16 genes. Gene expression was evaluated by reverse transcription-PCR and Western blot. The duration of the gene re-expressing effect of hydralazine was analyzed on T24 cells. Functionality of the re-expressed proteins was evaluated by the induction of the estrogen-responsive gene PS2 on MDA-231 cells and by the induction of G(1) arrest on T24 cells. The gene demethylating and re-expressing ability of hydralazine was tested in two patients with cervical and head and neck carcinomas, respectively. RESULTS: Hydralazine and procainamide induced de methylation and re-expression of the ER, RARbeta, and p16 genes in cultured cells. Both drugs also demethylated and re-expressed the ER gene in mice. Hydralazine re-expressed the p16 gene longer as compared with 5-aza deoxycytidine. The re-expressed genes were functional. In addition, the treatment with oral hydralazine demethylated and re-expressed the RARbeta and p16 genes in the cervical and head and cancer patients. CONCLUSIONS: These cardiovascular drugs have a promising tumor suppressor-reactivating action and could potentially be used in clinic as an anticancer treatment, most likely to increase the efficacy of current biological or chemotherapeutic treatments. PMID- 12738713 TI - Methylenetetrahydrofolate reductase polymorphism in advanced colorectal cancer: a novel genomic predictor of clinical response to fluoropyrimidine-based chemotherapy. AB - PURPOSE: Fluorouracil (5-FU) is widely used in the treatment of colorectal cancer. Methylenetetrahydrofolate reductase (MTHFR) could play an important role in the action of 5-FU, an inhibitor of thymidylate synthetase, by converting 5,10 methylenetetrahydrofolate, a substrate of thymidylate synthetase, to 5 methyltetrahydrofolate. A polymorphism in MTHFR (677 C-->T; A222V) reduces enzyme activity and presumably increases the level of 5,10-methylenetetrahydrofolate. This increase would be expected to correlate with an improved response to 5-FU. The aim of the present study was to investigate the association between the MTHFR polymorphism and response to 5-FU and other fluoropyrimidines in patients with metastatic colorectal cancer. EXPERIMENTAL DESIGN: Forty-three patients with metastatic colorectal adenocarcinoma were analyzed. All patients were treated with p.o. or i.v. fluoropyrimidine-based chemotherapy. A comprehensive chart examination was performed to determine tumor response rates. Genomic DNA was extracted from blood, and MTHFR genotypes were determined. RESULTS: At least one copy of the mutant valine allele was present in 26 patients (21 heterozygotes and 5 homozygotes). The remaining 17 patients carried only the alanine allele. Exploration of the relationship between MTHFR alleles and response rates revealed a statistically significant difference in the frequency of the valine allele among responders versus nonresponders (P = 0.0351). This observation was associated with an odds ratio of 2.86 (95% confidence interval 1.06-7.73) for a response in individuals with a valine allele. CONCLUSIONS: Our results show a link between the MTHFR polymorphism and tumor response to fluoropyrimidine-based chemotherapy and suggest that MTHFR genotyping may be of predictive benefit in selecting treatment regimens. PMID- 12738712 TI - Immunohistochemical expression of cyclooxygenase isoenzymes and downstream enzymes in human lung tumors. AB - PURPOSE: Prostanoids are important mediators of pulmonary vaso- and bronchotone regulation and strongly influence inflammatory reactivity. The product of cyclooxygenase (Cox), prostaglandin H(2), is further metabolized via downstream enzymes into the different effective metabolites. The specific cellular equipment with certain downstream enzymes crucially determines the cellular reactivity by generation of functionally different prostanoid metabolites. EXPERIMENTAL DESIGN: To elucidate the role of arachidonic acid metabolism via the cyclooxygenase pathway in different human lung tumors, expression of cyclooxygenase isoenzymes (Cox-1 and Cox-2) and downstream enzymes of prostanoid metabolism was investigated in human non-small cell lung cancer and normal human lung tissue by immunohistochemical techniques. RESULTS: In comparison to strong Cox-1 reactivity in airways and endothelial cells of normal lung specimens, only 4 of 15 adenocarcinomas showed infrequent Cox-1 expression. All lung cancer specimens displayed an increased Cox-2 immunostaining pattern with strong reactivity in adenocarcinomas and lower reactivity in squamous cell carcinomas. Adenocarcinomas and squamous cell carcinomas were also positive for thromboxane A(2) synthase, prostaglandin D(2) synthase, and prostaglandin E(2) synthase, but not for prostacyclin synthase. Endothelial cells of vessels found within or near the tumor show extensive immunostaining of Cox-2 and thromboxane A(2) synthase, whereas endothelial cells of normal lung specimens, in contrast, expressed Cox-1 and prostacyclin synthase. CONCLUSIONS: We conclude that non-small cell lung cancer shows a specific Cox-/downstream-enzyme expression pattern, which is specifically altered in lung tumor cells and tumor supplying vessels in contrast to normal lung tissue. This may have major impact on tumor progression and tumor associated inflammation via an altered prostanoid metabolism with consecutive tumor-associated blood flow distribution. PMID- 12738714 TI - Differential gene expression profiles in a human T-cell line stimulated with a tumor-associated self-peptide versus an enhancer agonist peptide. AB - PURPOSE: Previous studies have shown that a specific 9-mer amino acid epitope (designated CAP-1) of the human "self" tumor-associated carcinoembryonic antigen can be used to stimulate CD8+ T cells from peripheral blood mononuclear cells of carcinoma patients vaccinated with pox vector-based carcinoembryonic antigen vaccines. A T-cell receptor agonist epitope of CAP-1 (designated CAP1-6D) has been shown to enhance the stimulation of T cells over levels obtained using CAP 1. The purpose of this study was to analyze gene expression profiles in T cells stimulated with the native CAP-1 versus the agonist CAP1-6D peptide. EXPERIMENTAL DESIGN: Microarray analyses were conducted to analyze differential gene expression profiles of a T-cell line stimulated with native versus agonist peptides. RESULTS: Numerous genes and gene clusters are identified as differentially expressed as a consequence of stimulation with the agonist peptide versus the native peptide; two genes, however, stand out in magnitude: the chemokine lymphotactin and granzyme B. In particular, lymphotactin expression is >12 times more pronounced in agonist-stimulated T cells. An ELISA assay was developed that confirmed marked lymphotactin secretion in T cells when stimulated with the agonist versus the native peptide. A chemotaxis assay also demonstrated the biological activity of the lymphotactin produced. CONCLUSIONS: To our knowledge, these are the first studies of gene expression profiles of a defined T cell line in response to stimulation with a defined antigen. They are also the first to compare, via cDNA microarray, responses of a T-cell line to (a) a tumor associated self-antigen and (b) a native epitope versus an agonist epitope. PMID- 12738715 TI - Phase I study of TLK286 (glutathione S-transferase P1-1 activated glutathione analogue) in advanced refractory solid malignancies. AB - PURPOSE: The purpose of this study was to determine the dose-limiting toxicities (DLTs), the maximum tolerated dose, and the pharmacokinetics of the novel glutathione analog TLK286 administered by i.v. infusion. EXPERIMENTAL DESIGN: Patients with advanced malignancies received i.v. TLK286 administered as a 30-min constant rate infusion once every 3 weeks in escalating doses from 60 to 1280 mg/m(2). Patients underwent tumor assessment on day 43 and continued on treatment until disease progression or unacceptable toxicity. RESULTS: A total of 35 patients were treated with 109 cycles of TLK286. At 1280 mg/m(2), 3 of 5 patients developed one of two observed dose limiting toxicities (DLTs). The DLTs were: mild pancreatitis (1 of 5) and bladder symptoms (2 of 5) consisting of hematuria, dysuria, and urinary frequency. All of the patients with DLTs continued on TLK286 treatment at 960 mg/m(2) (one dose below maximum tolerated dose) without recurrence of DLTs. DLTs were transient, resolved without sequelae, and noncumulative. TLK286-related toxicities included grade 1-2 nausea, vomiting, fatigue, transient microscopic hematuria, and anemia. Of 31 evaluable patients, 10 patients continued therapy (median six cycles; range, four to nine cycles). Pharmacokinetic studies of TLK286 on cycle 1 revealed a mean elimination half life of 18 min (95% confidence interval, 16.1-19.9). Dose-proportional increases in both maximum blood concentrations and area under the blood-concentration-time curve were observed over the dose range of 60-960 mg/m(2). CONCLUSION: TLK286 was well tolerated in this study. TLK286 safety and pharmacokinetics support disease specific evaluations of TLK286 at doses <1280 mg/m(2) administered once every three weeks in the treatment of patients with advanced malignancies. PMID- 12738716 TI - A Phase I dose-escalation study of sibrotuzumab in patients with advanced or metastatic fibroblast activation protein-positive cancer. AB - PURPOSE: The purpose of this research was to determine the safety, immunogenicity, pharmacokinetics, biodistribution, and tumor uptake of repeat infusions of a complementarity-determining region grafted humanized antibody (sibrotuzumab) directed against human fibroblast activation protein (FAP). EXPERIMENTAL DESIGN: A Phase I open-label dose escalation study was conducted in patients with cancers epidemiologically known to be FAP positive. Patients were entered into one of four dosage tiers of 5, 10, 25, or 50 mg/m(2) sibrotuzumab, administered weekly for 12 weeks, with trace labeling with 8-10 mCi of (131)I in weeks 1, 5, and 9. RESULTS: A total of 26 patients were entered into the trial (15 males and 11 females; mean age, 59.9 years; age range, 41-81 years). Twenty patients had colorectal carcinoma, and 6 patients had non-small cell lung cancer. A total of 218 infusions of sibrotuzumab were administered during the first 12 weeks of the study, with 24 patients being evaluable. One patient received an additional 96 infusions on continued-use phase for a total of 108 infusions over a 2-year period, and 1 patient received an additional 6 infusions on continued use. There were no objective tumor responses. Only one episode of dose-limiting toxicity was observed. Therefore, a maximum tolerated dose was not reached. Treatment-related adverse events were observed in 6 patients during the infusional monitoring period. Four of the 6 patients, 3 of whom had associated positive serum human antihuman antibody, were removed from the study because of clinical immune responses. Gamma camera images of [(131)I]sibrotuzumab demonstrated no normal organ uptake of sibrotuzumab, with tumor uptake evident within 24-48 h after infusion. Analysis of pharmacokinetics demonstrated a similar mean terminal t(1/2) of 1.4-2.6 days at the 5, 10, and 25 mg/m(2) dose levels, and with a longer mean t(1/2) of 4.9 days at the 50 mg/m(2) dose level. CONCLUSION: Repeat infusions of the humanized anti-FAP antibody sibrotuzumab can be administered safely to patients with advanced FAP-positive cancer. PMID- 12738717 TI - Efficacy and toxicity of the angiogenesis inhibitor SU5416 as a single agent in patients with advanced renal cell carcinoma, melanoma, and soft tissue sarcoma. AB - INTRODUCTION: Vascular endothelial growth factor (VEGF) is a key regulator in angiogenesis. Preclinical and clinical data support the role of VEGF and angiogenesis in renal cell carcinoma (RCC), melanoma (M), and soft tissue sarcoma (STS). The tyrosine kinase inhibitor SU5416 is a potent inhibitor of the VEGF receptors 1 and 2. EXPERIMENTAL DESIGN: We investigated 145 mg/m(2) SU5416 twice weekly in patients with advanced or metastatic RCC, M, and STS. The primary objectives were efficacy and safety. Disease assessments were performed after 4 and 8 weeks of treatment and every 2 months thereafter. Documented stable disease (SD) lasting for > or =3 months was considered an antitumor response. RESULTS: A group of 29 patients was entered in the RCC trial, 20 patients in the M trial, and 31 patients in the STS trial. Response was observed in 6 (1 minor response and 5 SDs) of 24 evaluable patients (25%) in the RCC group, 6 (1 minor response and 5 SDs) of 26 patients (23%) in the STS group, and none of the patients in the M group. Progression-free survival ranged from 7 to 252 days (median 59 days) in the RCC group, from 7 to 260 days (median 60 days) in the STS group, and from 14 to 139 days (median 41 days) in the M group. Toxicities observed were those reported previously for SU5416. CONCLUSION: SU5416 single agent is well tolerated. The antitumor response was low in patients with RCC and STS, whereas no responses were seen in patients with M. PMID- 12738718 TI - Chronic lymphocytic leukemia-reactive T cells during disease progression and after autologous tumor cell vaccines. AB - PURPOSE: Tumor-reactive T cells were measured in patients with chronic lymphocytic leukemia (CLL) because vaccines that increase the activity of these cells might lead to better disease control. EXPERIMENTAL DESIGN: Proliferation and ELISPOT assays (for T cells producing IFN-gamma after stimulation by CD40 activated CLL cells) were used to determine the prevalence of tumor-reactive T cells in 25 CLL patients at various stages of disease progression. The effects of vaccines, composed of autologous-oxidized tumor cells, on both the clinical course and tumor-reactive T-cell numbers were then determined in 2 patients. RESULTS: CLL-reactive T cells were found at frequencies of > or =10(-3) in 6 of 11 patients. Significant proliferation was found in 15 of 25 patients and correlated with clinical stage. The inability to measure CLL-reactive T cells in the remaining patients was not uniformly a result of generalized T-cell dysfunction or defective antigen presentation by CD40-activated CLL cells. CLL reactive T-cell frequencies increased in response to vaccination with oxidized autologous tumor cells in a patient with preexisting CLL-reactive T cells but not in a patient where tumor-reactive T cells were undetectable in the ELISPOT assay. CONCLUSIONS: Tumor-reactive T cells exist in some CLL patients (mainly during earlier stages of disease) and may potentially mediate therapeutic responses if their numbers and activation states can be sufficiently increased by tumor vaccines. PMID- 12738719 TI - Phase II trial of tetrathiomolybdate in patients with advanced kidney cancer. AB - PURPOSE: Tetrathiomolybdate (TM), a copper-lowering agent, has been shown in preclinical murine tumor models to be antiangiogenic. We evaluated the antitumor activity of TM in patients with advanced kidney cancer in a Phase II trial. EXPERIMENTAL DESIGN: Fifteen patients with advanced kidney cancer were eligible to participate in this trial. TM was initiated p.o. at 40 mg three times a day with meals and 60 mg at bedtime to deplete copper. A target serum ceruloplasmin (CP) level of 5-15 mg/dl was defined as copper depletion. Doses of TM were reduced for grade 3-4 toxicity and to maintain a CP level in the target range. Once copper depletion was attained, patients underwent baseline tumor measurements and then again every 12 weeks for response assessment. Patients not exhibiting progressive disease at 12 weeks after copper depletion continued on treatment. Serum levels of Interleukin (IL)-6, IL-8, vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) were assayed pretreatment and at various time points on treatment. Dynamic contrast enhanced-magnetic resonance imaging (DCE-MRI) was performed on selected patients in an attempt to assess changes in tumor vascularity. RESULTS: All of the patients rapidly became copper depleted. Thirteen patients were evaluable for response. No patient had a complete response or PR. Four patients (31%) had stable disease for at least 6 months during copper depletion (median, 34.5 weeks). TM was well tolerated, with dose reductions most commonly occurring for grade 3-4 granulocytopenia of short duration not associated with febrile episodes. Serum levels of IL-6, IL-8, VEGF, and bFGF did not correlate with clinical activity. Serial DCE-MRI was performed only in four patients, and a decrease in vascularity seemed to correlate with necrosis of a tumor mass associated with tumor growth. CONCLUSIONS: TM is well tolerated and consistently depletes copper as measured by the serum CP level. Clinical activity was limited to stable disease for a median of 34.5 weeks in this Phase II trial in patients with advanced kidney cancer. Serum levels of proangiogenic factors IL-6, IL-8, VEGF, and bFGF may correlate with copper depletion but not with disease stability in this small cohort. TM may have a role in the treatment of kidney cancer in combination with other antiangiogenic therapies. PMID- 12738720 TI - Phase I evaluation of sequential topoisomerase targeting with irinotecan/cisplatin followed by etoposide in patients with advanced malignancy. AB - PURPOSE: To investigate pharmacologically guided addition of etoposide to a weekly irinotecan/cisplatin chemotherapy. PATIENTS AND METHODS: Patients with advanced nonhematologic malignancies were eligible. Treatment consisted of i.v. administration of 50 mg/m(2) irinotecan and 20 mg/m(2) cisplatin on days 1, 8, 15, and 22 of a 42-day cycle or on days 1 and 8 of a 21-day cycle. Etoposide was administered in a dose-escalating fashion 2 days after each dose of irinotecan/cisplatin, either i.v. as a single dose or p.o. as two doses administered 12 h apart. Pharmacologic analyses included measurement of plasma concentrations of irinotecan, SN-38, and SN-38 glucuronide, as well as quantitation of topoisomerase protein levels in peripheral blood mononuclear cells (PBMNCs). RESULTS: A total of 40 patients with a variety of malignancies received 122 cycles of therapy. Dose-limiting toxicities included neutropenia and diarrhea, with the 21-day cycle tolerated better than the 42-day cycle. For the 21-day cycle, the maximum tolerated dose was 75 mg/m(2) for i.v. etoposide and 85 mg/m(2) for oral etoposide. Objective responses were observed in four patients with previously treated mesothelioma, gastric, breast, and ovarian cancer, respectively. PBMNC levels of topoisomerase IIalpha were increased at the time of etoposide administration in two patients, with these patients having the highest SN-38 glucuronide peak-plasma-concentration and area-under-the-curve values among 15 patients with available pharmacokinetic data. One of these patients had a partial response to therapy. CONCLUSIONS: Pharmacologically guided administration of etoposide in combination with irinotecan/cisplatin using a 21-day cycle is associated with acceptable toxicity and significant antitumor activity. The finding that PBMNC topoisomerase IIalpha protein levels increased after irinotecan/cisplatin treatment in two of six patients supports the continued development of sequential topoisomerase targeting in the treatment of malignancy. PMID- 12738721 TI - Thalidomide metabolites in mice and patients with multiple myeloma. AB - PURPOSE: This research examines the profile of metabolites of thalidomide that are formed in refractory multiple myeloma patients undergoing thalidomide therapy in comparison with those that are detected in healthy mice. EXPERIMENTAL DESIGN: Urine or plasma samples from patients during thalidomide therapy (100-400 mg daily), or from mice treated i.p. (100 mg/kg) or p.o. with thalidomide (50 mg/kg) were analyzed using liquid chromatography-mass spectrometry. Metabolites in each of the peaks observed in the UV- and mass spectrometry-detected high-performance liquid chromatography traces were identified by comparison of retention times and spectra with those of authentic standards. RESULTS: Plasma and urine samples from mice 4 h after treatment with thalidomide contained eight major metabolites formed by hydroxylation and/or hydrolysis of thalidomide. In contrast, urine samples from seven multiple myeloma patients at steady state levels of thalidomide therapy showed the presence of only three hydrolysis breakdown products and no hydroxylated metabolites. CONCLUSIONS: Our results show that thalidomide metabolite profiles in multiple myeloma patients differ considerably from those in mice. The lack of measurable hydroxylated metabolites in urine and in 1 case plasma of these patients suggests that such metabolites are not responsible for the therapeutic effects of thalidomide in multiple myeloma. We suggest that thalidomide may act directly, down-regulating growth factors essential for multiple myeloma growth. PMID- 12738722 TI - Multicenter randomized Phase II study of paclitaxel (1-hour infusion), fluorouracil, hydroxyurea, and concomitant twice daily radiation with or without erythropoietin for advanced head and neck cancer. AB - PURPOSE: To expand on our experience with the combination of paclitaxel, fluorouracil, hydroxyurea, and twice daily irradiation (T-FHX) and to assess the impact of weekly administration of erythropoietin (r-HuEpo) on transfusion requirements, we conducted a Phase II multi-institutional trial with a simplified 1-h paclitaxel infusion schedule and randomized patients to receive weekly doses of r-HuEpo. PATIENTS AND METHODS: A total of 90 patients with locally advanced head and neck cancers (stage IV, 96%; N(2)/N(3), 66%) were treated on a regimen of 1-h infusion of paclitaxel (100 mg/m(2)/day, day 1), 120-h infusion of 5 fluorouracil (600 mg/m(2)/day, days 0-5); hydroxyurea 500 mg p.o. every 12 h for 11 doses; and radiation 150cGy bid, days 1-5 of each 14-day cycle repeated for five cycles over 10 weeks (7200-7500 cGy). Before initiating therapy, patients were randomized to receive r-HuEpo 40,000 IU s.c. once weekly. RESULTS: At median follow-up of 40 months, 3-year progression-free survival is 62%, locoregional control is 84%, and systemic control is 79%. Overall survival is 59%. Anemia, leucopenia, dermatitis, and mucositis were the most frequent grade 3 or 4 toxicities. Patients randomized to erythropoietin experienced less grade 2/3 anemia (52 versus 77%; P = 0.02), but transfusion requirements were not significantly different. CONCLUSIONS: T-FHX is an active and tolerable regimen inducing local tumor control and promising survival with organ preservation in high-risk patients. One h infusion of paclitaxel simplified the regimen without compromising efficacy. Addition of erythropoietin does not reduce the need for transfusion with this nonplatinum-containing regimen. T-FHX should be advanced to a randomized trial and compared with a cisplatin-based concomitant regimen. PMID- 12738723 TI - Induction docetaxel and carboplatin followed by weekly docetaxel and carboplatin with concurrent radiotherapy, then surgery in stage III non-small cell lung cancer: a Phase I study. AB - PURPOSE: To determine the maximum-tolerated dose of docetaxel (DOC) in combination with carboplatin (CAR) and thoracic radiotherapy (RT), in the setting of trimodality treatment of patients with stage III non-small cell lung cancer (NSCLC). EXPERIMENTAL DESIGN: Thirty-two patients with biopsy-proven stage IIIA (n = 20) or IIIB (n = 12) NSCLC were given two initial cycles of CAR (area under the curve = 6) and DOC (75 mg/m(2)), subsequent RT (54 Gy) with concurrent weekly CAR (area under the curve = 2), and DOC at six dose levels from 10 to 40 mg/m(2), then surgery if the patient's disease was resectable. RESULTS: Three patients did not complete induction computed tomography (CT). Twenty-nine patients received concurrent CT/RT. Fifteen patients were eligible for surgery. Dose-limiting toxicities occurred in 2 patients, at dose levels two (atrial fibrillation) and three (transaminitis). The maximum-tolerated dose, as defined by the protocol, was not reached, although grade 3 and 4 toxicities were encountered at all dose levels. The most common more than or equal to grades 3 toxicities were neutropenia, nausea, vomiting, and fatigue. Four patients (13.3%) responded to induction CT. Ten patients (38.5%) responded to CT/RT. Eight surgical patients (57.1%) were downstaged, including 3 pathologic complete responses. Median relapse free and overall survivals are 8.5 and 12 months. One-year and estimated 2-year survival rates are 56.3 and 34.3%. CONCLUSION: This new regimen for stage III NSCLC of induction CAR/DOC, then weekly CAR/DOC with concurrent RT followed by surgery, can be safely administered and offers encouraging results. DOC at 30 mg/m(2) in combination with CAR and RT is recommended for Phase II study. PMID- 12738725 TI - Differential splicing of KLK5 and KLK7 in epithelial ovarian cancer produces novel variants with potential as cancer biomarkers. AB - PURPOSE: The wild-type or variant mRNAs of several kallikrein (KLK) genes, such as KLK4, are highly expressed in ovarian carcinomas and may have potential as tumor markers. Two of these KLK genes (KLK5 and KLK7) and their proteins (hK5 and hK7) were first identified in the skin epidermis, where hK5 may be the physiological activator of hK7. The purpose of this study was to reexamine the expression of KLK5/hK5 and KLK7/hK7 and their association and to determine whether cancer-related variant transcripts were expressed. EXPERIMENTAL DESIGN: The expression of KLK5/hK5 and KLK7/hK7 was analyzed in the same cohort (n = 37) of benign (n = 4) and malignant ovarian tissue (n = 23) samples and primary cultured cells (n = 21) and in 8 ovarian cancer cell lines using semiquantitative RT-PCR; Southern, Northern, and Western blot analyses; and immunohistochemistry techniques. RESULTS: We showed the concordant higher expression of both KLK5/hK5 and KLK7/hK7 in ovarian carcinomas, especially late-stage serous carcinomas, compared with normal ovaries and benign adenomas. We also found that one novel KLK5 transcript with a short 5'-untranslated region and a novel KLK7 transcript with a long 3'-untranslated region were highly expressed in the ovarian cancer cell lines OVCAR-3 and PEO1, respectively, but were expressed at very low levels in normal ovarian epithelial cells. Both Western blot and immunohistochemistry analyses showed that these two enzymes are secreted from ovarian carcinoma cells. CONCLUSIONS: Our study demonstrated that hK5 and hK7, or more specifically, the short KLK5 and long KLK7 transcripts, may be useful as tumor markers for epithelial-derived serous carcinomas. However, additional clinical studies assessing serum levels of these putative biomarkers are required to confirm their usefulness in the diagnosis and/or monitoring of these tumors. PMID- 12738724 TI - Cytochrome p450 and glutathione transferase expression in human breast cancer. AB - PURPOSE: The cytochrome P-450 (CYP) and glutathione S-transferase (GST) enzyme systems may influence the biological effects of carcinogens, including estrogens. As such, these enzymes may predict the developmental risk of breast cancer, as well as be potential targets for chemoprevention. The purpose of this study was to compare the expression of GST-Pi and CYPs 1A1, 2B6, 2E1, and 3A4 in paired samples of normal and malignant breast tissue from patients with breast cancer and women undergoing reduction mammoplasty. EXPERIMENTAL DESIGN: Expression of CYPs 1A1, 2B6, 2E1, 3A4, and GST-Pi was quantified in breast tissue from 33 patients with breast cancer and in 17 women without history of cancer who underwent reduction mammoplasty. The expression of CYP 1A1, 2B6, 2E1, 3A4, and GST-Pi was quantified by immunoblotting. RESULTS: CYP 1A1, 2E1, and 3A4 expression was significantly lower (P < 0.05) in malignant tissue as compared with morphologically normal adjacent tissue. Conversely, GST-Pi expression was marginally lower in the normal tissue (P = 0.08). No significant difference in enzyme expression was seen between the tissue from reduction mammoplasty and normal tissue from breast cancer patients. There was a trend for higher expression of CYP 2B6 and GST-Pi in the estrogen receptor expressing tumors than those tumors without expression (P > 0.28). CONCLUSION: The expression of these enzymes was similar in morphologically normal breast tissue from patients with or without breast cancer. The expression of CYPs was down-regulated in the tumor tissue. The clinical significance of CYP alterations in breast cancer will need further characterization. PMID- 12738727 TI - Serum CYFRA 21-1 in advanced stage non-small cell lung cancer: an early measure of response. AB - OBJECTIVES: Our objective was to test the prognostic importance of both the pretreatment level and change in serum CYFRA 21-1 after one cycle of chemotherapy in patients with advanced non-small cell lung cancer (NSCLC) and to compare these two CYFRA variables to routine clinical stage and response as measured by imaging. PATIENTS AND METHODS: Our patients consisted of 58 with advanced NSCLC who were treated with chemotherapy. Fourteen were stage IIIa, 8 stage IIIb, and 36 stage IV, and none had received previous treatment. The choice of chemotherapy was left to the discretion of the treating physicians. We collected two serum samples, one before the first cycle of chemotherapy and the second before the second cycle, and analyzed these for serum CYFRA 21-1 using an electrochemiluminescence immunoassay and the ElecSys 2010 system (Roche Diagnostics Corp., Indianapolis, IN). We expressed changes in CYFRA in terms of the natural ratio logarithm of post-treatment to pretreatment CYFRA, and we used the Cox proportional hazards model to analyze survival time. RESULTS: Patients experienced an average drop of 27% in serum CYFRA after the first cycle of chemotherapy. Furthermore, the Cox model demonstrated that both the initial natural logarithm of serum CYFRA and presence of >27% drop in CYFRA were significantly related to subsequent survival (model P < 0.0006), but neither clinical stage nor clinical response related to survival (P > 0.1). CONCLUSION: In advanced stage NSCLC, the initial level of serum CYFRA appears to provide more prognostic information than clinical stage. Furthermore, a drop of >27% in CYFRA after one cycle of therapy adds prognostic information, so that this threshold appears to be an early measure of response to chemotherapy. PMID- 12738726 TI - Characterization of human lymphocyte antigen class I antigen-processing machinery defects in renal cell carcinoma lesions with special emphasis on transporter associated with antigen-processing down-regulation. AB - The HLA class I antigen-processing machinery (APM) plays a crucial role in the generation of peptides from endogenously synthesized proteins and in their presentation to cytotoxic T lymphocytes. The potential role of defects of APM components in immune escape mechanisms used by malignant cells has prompted us to analyze their expression in renal cell carcinoma (RCC) lesions with special emphasis on TAP because of its critical role in the loading of HLA class I antigens with peptides. Immunohistochemical staining of 51 formalin-fixed RCC lesions and autologous normal renal epithelium detected transporter associated with antigen processing (TAP)1 and tapasin deficiencies in 63 and 80% of the tumor lesions. Impaired low molecular weight protein (LMP)2 and LMP7 expression was found in 73 and 33% of the RCC lesions analyzed, respectively. In contrast to the high frequency of APM component down-regulation, HLA class I heavy chain and beta(2)-microglobulin defects were detected in only 12 and 10% of the lesions, respectively. Concomitant TAP1 and LMP2 deficiencies were found in approximately 57% of RCC lesions, whereas a coordinated down-regulation of all APM components occurred only in 5% of the tumor specimens analyzed. The presence of APM defects was independent of tumor stage and grade but varied significantly among the RCC subtypes. TAP abnormalities do not appear to be attributable to structural alterations because no mutations in TAP1 were detected in TAP1-deficient RCC lesions. These data suggest that TAP defects in RCC lesions are caused by regulatory abnormalities. Therefore, T-cell-based immunotherapy may benefit from the administration of cytokines that up-regulate TAP expression. PMID- 12738728 TI - Expression of thrombospondin-derived 4N1K peptide-containing proteins in renal cell carcinoma tissues is associated with a decrease in tumor growth and angiogenesis. AB - PURPOSE: We have reported that the 4N1K peptide (KRFYVVMWKK) from thrombospondin (TSP) 1 has antiangiogenic activities. The goal of this study was to examine whether the expression of 4N1K-containing proteins correlates with reduced growth of human renal cell carcinoma (RCC). EXPERIMENTAL DESIGN: We examined the expression of 4N1K-containing proteins and TSP1, microvessel density, proliferation index, and apoptotic index in 119 surgically excised RCC tissues by immunohistochemical techniques. The correlation between the above variables and clinicopathological features was analyzed statistically. RESULTS: The antibody raised against the 4N1K peptide recognized protein fragments of matrix metalloproteinase 3-digested TSP1 and positively stained the sections of renal cancer tissues. These reactions disappeared when the antibody was preincubated with immobilized 4N1K peptide, suggesting that the reactions were 4N1K peptide specific. Although TSP1 expression did not correlate with various clinicopathological features and tumor size, all 4N1K-positive tumors were locally confined and of significantly smaller size (median, 3.3 cm; range, 2.0 4.4 cm) than 4N1K-negative tumors (median, 5.2 cm; range, 2.8-8.8 cm; P < 0.001). 4N1K-positive tumors exhibited significantly lower microvessel density and higher apoptotic index of tumor cells than 4N1K-negative tumors (P < 0.001 and P = 0.042, respectively). CONCLUSIONS: Our data suggest that expression of 4N1K containing proteins in tumor tissues is associated with reduced angiogenesis and tumor growth; thus, it would be a potentially predictive marker for progression of RCC. PMID- 12738729 TI - Expression of cyclooxygenase-2 in renal cell carcinoma: correlation with tumor cell proliferation, apoptosis, angiogenesis, expression of matrix metalloproteinase-2, and survival. AB - PURPOSE: Cyclooxygenase (COX)-2 plays an important role in tumor cell proliferation, resistance to apoptosis, angiogenesis, and invasion in various malignant tumors. However, the relationships between COX-2 expression and these biological processes, clinicopathological features, and survival rate in patients with renal cell carcinoma are not clear. EXPERIMENTAL DESIGN: Tumor sections surgically removed from 131 patients were examined for COX-2 expression by immunohistochemistry. We also examined Ki-67 labeling index, apoptotic index, microvessel density, and matrix metalloproteinase (MMP)-2 expression, and correlated COX-2 expression with various clinicopathological features and survival. RESULTS: Of 131 sections, 70 (53.4%) were positive for COX-2 expression. COX-2 expression was associated significantly with various clinicopathological features, and correlated with the Ki-67 labeling index, microvessel density, and MMP-2 expression (P < 0.01), but not with the apoptotic index (P = 0.054). COX-2 expression was also identified as an independent risk factor for large tumor size (>7 cm) in multivariate logistic regression model. COX proportional hazards analysis showed that distant metastasis and high T stage were independent prognostic factors [odds ratio (OR), 9.41; 95% confidence interval (CI), 2.16-41.11; P < 0.01 and OR, 5.19; 95% CI, 1.02-26.54; P = 0.048, respectively), whereas COX-2 expression was not (OR, 1.46; 95% CI, 0.24-9.00; P = 0.68). CONCLUSION: COX-2 expression in patients with renal cell carcinoma is associated with several clinicopathological factors, and appeared to play an important role in tumor cell proliferation and MMP-2 expression, but is not a significant prognostic factor. PMID- 12738730 TI - The gene ratios c-MYC:cyclin-dependent kinase (CDK)N2A and CCND1:CDKN2A correlate with poor prognosis in squamous cell carcinoma of the head and neck. AB - PURPOSE: Tumor-Node-Metastasis classification does not fully predict outcome of treatment and prognosis in patients with squamous cell carcinoma of the head and neck. Different biomarkers have been suggested to yield additional prognostic information, but no single marker has thus far been introduced in the clinic. The objective of the present study was to analyze the copy number of the frequently amplified oncogenes CCND1 and c-MYC in relation to the commonly deleted tumor suppressor gene cyclin-dependent kinase (CDK)N2A (p16) to enhance the clinical significance. EXPERIMENTAL DESIGN: Extracted DNA from diagnostic biopsies of 78 untreated patients were analyzed by real-time PCR with specific primers for c MYC, CCND1, and CDKN2A. Gene copy number ratios were calculated by dividing the copy number of c-MYC or CCND1 with CDKN2A. Ratios > 2 were defined as enhanced. These data were related to disease-free interval and disease-specific survival. RESULTS: Enhanced gene ratio of c-MYC:CDKN2A was detected in 35 of 78 (45%) and enhanced ratio of CCND1:CDKN2A in 36 of 78 (46%) of the cases. The c-MYC:CDKN2A and CCND1:CDKN2A ratios correlated with disease-specific survival with respect to death (P = 0.042 and 0.049, respectively; Log-rank test). Furthermore, enhancement of c-MYC:CDKN2A was associated with a shorter disease-free interval as marked by the development of recurrences or metastases (P = 0.014; Log-rank test). CONCLUSIONS: We conclude that CCND1 and/or c-MYC amplification, when combined with CDKN2A deletion, yield additional prognostic information as compared with analysis of single genetic aberrations. These gene ratios, as analyzed by a sensitive method like real-time PCR on diagnostic biopsies, might help clinicians to individualize the treatment of squamous cell carcinoma of the head and neck as they reflect the biological properties of the tumors. This could be used as an adjunct to the Tumor-Node-Metastasis classification system. PMID- 12738731 TI - Skp2 overexpression is a prognostic factor in patients with ovarian adenocarcinoma. AB - PURPOSE: The purpose of this study was to examine Skp2 expression in epithelial ovarian tumors and to identify the association of Skp2 expression levels with patient survival. EXPERIMENTAL DESIGN: Skp2 protein expression was examined by immunohistochemistry in 134 epithelial ovarian tumors [20 adenomas, 23 low malignant potential (LMP) tumors, and 91 adenocarcinomas]. Results of immunostaining were correlated with clinicopathological variables and overall survival. Skp2 mRNA expression was examined by semiquantitative PCR in 32 ovarian adenocarcinomas and in 3 ovarian cancer cell lines. RESULTS: Skp2 expression was detected in neither ovarian adenomas nor LMP tumors. In contrast, Skp2 expression was detected in 47.3% (43 of 91) of adenocarcinomas. Positive Skp2 expression was detected significantly more often in adenocarcinomas than in LMP tumors (P < 0.0001) or in adenomas (P < 0.0001). A significantly higher detection rate of Skp2 expression was observed in advanced-stage diseases compared with early-stage diseases (P = 0.010). Log-rank testing showed that Skp2 overexpression was significantly correlated with poor patient survival (P = 0.0035). Older age (P = 0.0026), advanced clinical stage (P < 0.0001), and high histological grades of the tumors (P = 0.0018) were also significantly associated with poor prognoses. In multivariate analysis, Skp2 overexpression (P = 0.0069) and clinical stage (P < 0.0001) remained significantly associated with overall survival, whereas age and histological grade lost their significance. Considerable levels of Skp2 mRNA expression were detected in all ovarian adenocarcinomas examined by semiquantitative PCR. CONCLUSIONS: Skp2 expression might play an important role in the development and progression of ovarian adenocarcinomas, and Skp2 overexpression is an independent prognostic marker of ovarian adenocarcinoma patients. PMID- 12738732 TI - Overexpression of CDC25A phosphatase is associated with hypergrowth activity and poor prognosis of human hepatocellular carcinomas. AB - PURPOSE AND EXPERIMENTAL DESIGN: CDC25 genes are cell cycle-activating phosphatases that positively regulate the activity of cyclin-dependent kinase. CDC25A and CDC25B, being oncogenes, are overexpressed in a variety of human malignancies. To investigate the potential roles of CDC25s in hepatocellular carcinoma (HCC), expression of CDC25A and CDC25B was examined in human HCC samples. RESULTS: Reverse transcription-PCR showed that overexpression of CDC25A and CDC25B mRNAs was found in 9 of 13 (69%) and 4 of 13 (31%) HCCs, respectively. Immunohistochemistry of 59 HCCs showed marked increase in CDC25A expression, but not CDC25B, in HCC compared with noncancer tissues, and high expression of CDC25A in 33 of 59 (56%) HCCs. Overexpression of CDC25A in HCC was confirmed by Western blot analysis. High expression of CDC25A was associated with dedifferentiated phenotype and portal vein invasion (P = 0.001 and 0.031, respectively), and expression of CDC25A correlated well with proliferating cell nuclear antigen labeling index (P = 0.005). Univariate analysis indicated that high expression of CDC25A and proliferating cell nuclear antigen were both significant predictive factors for shorter disease-free survival (P = 0.004 and 0.039, respectively). Multivariate analysis indicated that CDC25A was an independent prognostic marker for disease-free survival (risk ratio for cancer relapse, 2.98; P = 0.029), even when analyzed with several clinicopathologic factors. On the other hand, expression of CDC25B did not correlate with any clinicopathological features. CONCLUSION: Our findings suggest that CDC25A, but not CDC25B, could be used as an independent prognostic marker for HCC. Our data would also contribute to forward understanding of tumor biology of HCC that is associated with cell cycle regulation. PMID- 12738733 TI - Promoter methylation of INK4a/ARF as detected in bile-significance for the differential diagnosis in biliary disease. AB - PURPOSE: There is a need to enhance endobiliary cytotechniques by molecular marker lesions. This is of special significance for patients with primary sclerosing cholangitis, a disease predisposing for the development of cholangiocarcinoma. The INK4a/ADP ribosylation factor (ARF) locus encodes two tumor suppressor genes: p16INK4a and p14ARF. p16INK4a has been shown to be of major significance in cholangiocarcinoma. EXPERIMENTAL DESIGN: In an effort to evaluate the potential diagnostic role of p16INK4a and p14ARF promoter methylation in biliary disease, endoscopical obtained bile specimens of 71 patients were analyzed (26 choledocholithiasis, 6 with normal results, 23 bile duct carcinoma, 5 gall bladder carcinoma). Eleven patients with primary sclerosing cholangitis were enrolled. RESULTS: Merely 6% of specimens (2 of 32) obtained from patients without evidence for malignant biliary disease but 53.5% of malignancies (15 of 28) showed p16 promoter methylation (p14: 3 and 46.2%, respectively). The concordance of methylation rates detected in either bile or tissue specimens was high. In primary sclerosing cholangitis, a similar prevalence of methylation was detected as in malignant disease. CONCLUSIONS: This study demonstrates: (a) a high frequency and specificity of INK4a/ARF methylation in malignant biliary disease compared with mere cholangitis; and (b) the capability to detect these alterations reliably in endoscopically obtained bile. Thus, INK4a/ARF's promoter methylation status represents a candidate marker for the endoscopic diagnosis of biliary disease. PMID- 12738735 TI - Lack of interleukin-10 expression could predict poor outcome in patients with stage I non-small cell lung cancer. AB - PURPOSE: Interleukin-10 (IL-10) may play an important role in controlling tumor growth and metastasis. Some reports have shown that IL-10 can be a potent inhibitor of tumor growth, but others suggest that IL-10 expression by the tumor is an adverse prognostic factor. Because normal bronchial epithelial cells constitutively produce IL-10, we decided to test the prognostic value of IL-10 in a well-defined population of patients with stage I non-small cell lung cancer (NSCLC) treated in a single institution. PATIENTS AND METHODS: Using immunohistochemical analysis, we retrospectively analyzed IL-10 expression in specimens from 138 patients with completely resected clinical/radiographic stage I NSCLC for whom clinical follow-up data were available. RESULTS: IL-10 expression was retained (IL-10 labeling index > or = 10%) in 94 patients (68.1%) and lost in 44 patients (31.9%). The duration of overall, disease-specific, and disease-free survival in the 44 patients lacking IL-10 expression was worse than in the 94 patients with IL-10 expression (P = 0.08, 0.02, and 0.05, respectively; Log-rank test). Interestingly, IL-10 expression was observed more frequently in tumors with squamous cell histology than in tumors of other histological subtypes (P = 0.04; chi(2) test). Multivariate analysis confirmed the independent prognostic value of IL-10 expression for disease-specific survival (P = 0.04). CONCLUSION: Lack of IL-10 expression by the tumor was associated with a significantly worse outcome of early stage NSCLC. The mechanisms underlying this clinically and biologically important finding need to be further explored. PMID- 12738734 TI - RECK expression in pancreatic cancer: its correlation with lower invasiveness and better prognosis. AB - BACKGROUND: The reversion-inducing cysteine-rich protein with Kazal motifs (RECK) gene was initially isolated as a transformation suppressor gene. The RECK gene is expressed widely in normal organs but is undetectable in many tumor-derived cell lines. When artificially expressed in such cell lines, RECK negatively regulates at least matrix metalloprotease (MMP)-9, MMP-2, and MT1-MMP activation and suppresses the invasive and metastatic potentials of these cells. Clinical relevance of these observations, however, is yet to be established. The aim of this study was to examine RECK expression in pancreatic cancer, where intensive invasiveness and metastasis are frequently observed, and investigate its clinical significance. We also analyzed the correlation between RECK expression and MMP activation. METHODS: (a) RECK expression in surgically resected tissue samples of invasive ductal carcinomas of the pancreas (n = 50) was examined immunohistochemically, and its correlation with clinicopathological factors was analyzed; and (b) gelatin zymography was used for the detection of latent and activated forms of MMP-2 and MMP-9 in some of the tissue samples (n = 33). The gelatinase activity was quantified by densitometory, and the ratio of intensity of the active MMP-2 band to the total intensity of the pro- and active MMP-2 bands was evaluated as an indicator of MMP-2 activation. The MMP-9 activation was also studied. RESULTS: Among the 50 ductal carcinoma samples, 26 (52%) were stained positive for RECK. In the normal pancreas, both acinar and beta cells were stained positive, but ductal cells did not. Tumors with positive RECK staining were significantly less invasive as compared with RECK-negative tumors (P = 0.0438). Importantly, patients who had tumors with high RECK expression showed significantly better prognosis than those who had RECK-negative tumors (P = 0.0463, by Log-rank test). Zymographic analysis indicated significant inverse correlation between the level of RECK expression and extent of MMP-2 activation (P = 0.0374). CONCLUSIONS: Our findings support the hypothesis that the RECK protein has negative effects on the invasiveness of pancreatic cancer by inhibiting MMP-2 activation and suggest the potential value of RECK as a prognostic molecular marker for pancreatic cancer. PMID- 12738736 TI - Classification of follicular thyroid tumors by molecular signature: results of gene profiling. AB - PURPOSE: Thyroid nodules are common, with a lifetime risk of developing a clinically significant thyroid nodule of 10% or higher. Preoperative diagnosis was greatly enhanced by the introduction of fine needle aspiration in the 1970s, but there has been little advancement since that time. Discrimination between benign and malignant follicular neoplasms is currently not possible by fine needle aspiration and can even be difficult after full pathologic review. The purpose of these studies is to identify genes expressed in follicular adenomas and carcinomas of the thyroid that will permit molecular differentiation of these neoplasms. EXPERIMENTAL DESIGN: Gene expression patterns of 17 thyroid follicular tumors were analyzed by oligonucleotide array analysis. Gene profiles for follicular adenomas and carcinomas were identified, and the two groups were compared for differences in expression levels. The differentially expressed genes were used to perform a hierarchical clustering analysis training set. Five follicular tumors with diagnosis undisclosed to the investigators and 2 minimally invasive carcinomas were entered into the cluster analysis as a test set to determine whether diagnosis by gene profile correlated with that obtained by pathologic evaluation. RESULTS: Thyroid follicular adenomas and carcinomas showed strikingly distinct gene expression patterns. The expression patterns of 105 genes were found to be significantly different between follicular adenoma and carcinoma. Many uncharacterized genes contributed to the distinction between tumor types. For five follicular tumors for which the final diagnosis was undisclosed, the clustering algorithm gave the correct diagnosis in all 5 cases. CONCLUSIONS: Gene profiling is a useful tool to predict the molecular diagnosis of follicular thyroid tumors. Genes were identified that reliably differentiate follicular thyroid carcinoma from adenoma. This study provides insight into genes that may be important in the molecular pathogenesis of follicular thyroid tumors, as well candidates for preoperative diagnosis of follicular thyroid carcinoma. PMID- 12738737 TI - Beta-catenin-related anomalies in apoptosis-resistant and hormone-refractory prostate cancer cells. AB - PURPOSE: beta-Catenin is a critical end component of the wnt signaling pathway that regulates cell growth, apoptosis, and migratory behavior in response to intercellular adhesion molecules. The aim of this study was to evaluate abnormalities of beta-catenin protein expression, subcellular localization, and activity in an in vitro model of acquired apoptosis-resistance in cultured PC cells and in primary human prostate cancers (PrCa). EXPERIMENTAL DESIGN: Apoptosis-resistant human prostate cancer cell line variants were derived from parental LNCaP cells by repeated brief exposure to apoptotic stimuli. The derivative and parental cells were analyzed for beta-catenin expression and intracellular localization using cell fractionation and Western blotting procedures. Endogenous transcriptional activity from the TCF/LEF-1 response element was also studied in these variants after transfection with a beta-catenin sensitive reporter plasmid. Finally, beta-catenin protein expression and intracellular localization were evaluated on 212 patients [122 localized PrCa and 90 hormone-refractory (HRPC) PrCa specimens by immunohistochemistry]. RESULTS: Western blot analysis showed that the intracellular partitioning of beta-catenin was shifted from the membrane fraction in parental cells to the cytoplasmic/nuclear fractions of the apoptosis-resistant cell lines. Coordinately, transcriptional activity from a TCF/LEF-promoted reporter plasmid was increased significantly in the apoptosis-resistant lines. In the primary prostate tumors analyzed, cytoplasmic and/or nuclear beta-catenin expression was correlated statistically with the HRPC status and Gleason score. In the group of localized PrCa, abnormal beta-catenin expression tended to be associated with a higher Gleason score and with pT3 disease. No mutation was found in patients with HRPC and abnormal beta-catenin expression. CONCLUSION: These data suggest that anomalies of beta-catenin expression occur in PrCa and that these anomalies are associated with disease progression, especially to the therapeutic-resistant state. PMID- 12738738 TI - Gain of chromosome 8q23-24 is a predictive marker for lymph node positivity in colorectal cancer. AB - PURPOSE: The prognosis of patients with colorectal cancer is largely determined by tumor stage. In this respect, colorectal cancers with lymph node metastases indicate a worse prognosis versus lymph node-negative tumors. Accordingly, there is considerable clinical interest in understanding the genetic mechanisms underlying metastasis formation. Furthermore, sensitive and specific biomarkers are needed to predict the metastatic phenotype at the time of diagnosis. EXPERIMENTAL DESIGN: Fifty colorectal cancers with or without lymph node metastases were assessed for genomic imbalances by comparative genomic hybridization. Particular interest was focused on whether specific chromosomal alterations exist in primary tumors that might be indicative and specific for the metastatic phenotype. RESULTS: The analysis revealed that lymph node-positive colorectal cancers show a higher degree of chromosomal instability than lymph node-negative cancers (average number of chromosomal copy alterations, 9.8 versus 7.5). Chromosomal alterations commonly described in colorectal cancers such as gain of 20q or loss of 18q21 were not different. However, the gain of chromosomal region 8q23-24 was seen in the vast majority of lymph node-positive cancers, whereas it was rather rare in lymph node-negative carcinomas (P = 0.0016). CONCLUSIONS: These data suggest that genes located at 8q23-24 might favor the development of lymphatic metastases in colorectal cancers. Additionally, the gain of this region could be used to predict the metastatic potential of primary colorectal cancers. PMID- 12738739 TI - The loss of 5alpha-reductase type I and type II mRNA expression in metastatic prostate cancer to bone and lymph node metastasis. AB - PURPOSE: Considerable evidence has accumulated demonstrating that the 5alpha reduction of testosterone to dihydrotestosterone occurs more efficiently in the normal and benign hyperplastic prostate than in prostate cancer tissues. Efforts have also been channeled into investigating the distribution of 5alpha-reductase isoenzymes in primary prostate tissues and in "in vitro" cell models of the human prostate. However, no one has, thus far, examined the expression of these isoenzymes in prostate cancer metastasis, although such studies might shed some light on the mechanism(s) responsible for the loss of hormone sensitivity in those tumors. The present report addresses this issue in the hope that this might help to identify the steps leading to the development of prostate cancer metastasis. EXPERIMENTAL DESIGN: In the present study we used in situ mRNA hybridization of sections from archival paraffin-embedded material to investigate the expression of 5alpha-reductase type I (5alphaR-I) and type II (5alphaR-II) mRNAs in prostate cancer bony (n = 9) and lymph node (n = 13) metastasis, and compared the mRNA distributions with those observed in sections from primary prostate tumors (n = 12). In parallel, sections were investigated for androgen receptor (AR) mRNA expression, and immunostained for AR and prostate-specific antigen. RESULTS: Neither 5alphaR-I nor 5alphaR-II mRNA expression was detected in any of the prostate metastatic lesions, although the same metastatic sites expressed AR mRNA, and stained for cytoplasmic prostate-specific antigen and nuclear AR. In contrast, primary prostate tumors displayed intense staining for 5alphaR-I and 5alphaR-II. CONCLUSION: These findings suggest that the loss of 5alpha-reductase mRNA expression in bone and lymph node metastasis may be associated, in part, with the progression of these tumors to androgen insensitivity. PMID- 12738740 TI - Stroma cells: a novel target of herceptin activity. AB - PURPOSE: Stroma cells play a relevant role in tumor development and progression. We investigated the activity of herceptin (HER), a humanized monoclonal antibody widely used for the treatment of HER2-overexpressing epithelial cancer, toward stroma cell lines L87/4 and L88/5. EXPERIMENTAL DESIGN: We studied the antiproliferative potential of HER and role of human serum in HER activity. We also investigated the ability of HER to alter ancillary functions of L87/4 and L88/5, such as support to long-term hematopoiesis, growth factor production, breast cancer cell adhesion, and proliferation. RESULTS: Flow cytometry showed that HER2 membrane expression in L87/4 and L88/5 stroma cells was intermediate between the expression in HER2-negative/dim MCF-7 breast cancer cells and HER2 bright SK-BC3 breast cancer cells. HER2 gene amplification was not detected by fluorescence in situ hybridization in either stromal cell lines. HER significantly inhibited L87/4 and L88/5 proliferation. Mean ID(50)s were found to be 2000 and 1700 micro g/ml for L87/4 and L88/5, respectively, after 3-day exposure and 800 micro g/ml for both cell lines after 9-day exposure. The presence of 10% human serum in the culture increased HER inhibitory activity. IC(50) of stroma cells was found to be intermediate between HER2-bright breast cancer cells (SK-BC3) and HER2-negative/dim breast cancer cells (MCF-7). The drug did not significantly affect the ability of stroma cells to support long-term hematopoiesis in the cobblestone area forming cell assay. In contrast, in coculture assay, MCF7 cells demonstrated a worse adhesion and growth capability on HER-treated stroma layers when compared with untreated stroma. Moreover, HER significantly reduced vascular endothelial growth factor production by L88/5 cells. CONCLUSIONS: Our data support the novel finding that HER may have a relevant activity against stroma cells. PMID- 12738741 TI - Effect of interleukin (IL)-4 cytotoxin on breast tumor growth after in vivo gene transfer of IL-4 receptor alpha chain. AB - Although human breast cancer cells express interleukin-4 receptors (IL-4Rs), a recombinant fusion protein, IL-4 cytotoxin, did not mediate desirable antitumor activity in tumor models of breast cancer. Recent studies have identified that a primary IL-4 binding protein, IL-4Ralpha chain, is internalized after binding to IL-4 in cancer cells. The consequent expression of high-level IL-4Ralpha in tumor cells sensitizes them to the cytotoxic effect of IL-4 cytotoxin in vitro. To assess whether overexpression of IL-4Ralpha chain in vivo by plasmid-mediated gene transfer can enhance antitumor activity of IL-4 cytotoxin in mouse models of breast tumor, we injected MDA-MB-231 human breast cancer cells in both flanks of athymic nude mice. Animals then received three intratumoral (i.t.) injections of either IL-4Ralpha encoding vector (left flank) or vector only (right flank) mixed with liposome followed by IL-4 cytotoxin administration. Both i.p. and i.t. administration of IL-4 cytotoxin profoundly reduced the growth of IL-4Ralpha plasmid-injected MDA-MB-231 tumors, compared with control. Innate immune cells, including macrophages and neutrophils, were found to infiltrate at the regressing tumor site. This study provides proof of principle that i.t. IL-4Ralpha plasmid injection followed by systemic or i.t. IL-4 cytotoxin administration may be a useful strategy for the treatment of breast cancer. PMID- 12738742 TI - Vaccine therapy of established tumors in the absence of autoimmunity. AB - PURPOSE: Many current clinical trials involve vaccination of patients with vaccines directed against tumor-associated antigens, which are, in actuality, "self-antigens" overexpressed in tumors as compared with normal tissues. As tumor vaccines become more potent through the addition of costimulatory molecules and cytokines and the use of diversified prime and boost regimes, the level of concern rises regarding the balance between antitumor immunity and pathological autoimmunity. Studies were conducted using mice bearing a transgenic self-antigen [human carcinoembryonic antigen (CEA)], which is expressed in some normal adult tissues, and tumor expressing the same self-antigen. These mice were vaccinated with recombinant poxviral vectors [recombinant vaccinia, recombinant fowlpox (rF)] encoding the CEA transgene as well as a triad of costimulatory molecules [B7-1, ICAM-1, and LFA-3 (TRICOM)]. Here we investigate the mechanism of tumor therapy and evaluate the safety of such a regimen in a self-antigen system. To our knowledge, the study reported here is the first description of a vaccine to a defined antigen where the regimen is potent enough to induce tumor therapy in the absence of autoimmunity. EXPERIMENTAL DESIGN: CEA transgenic mice were transplanted with CEA-expressing tumors. Fourteen days later, mice were vaccinated with recombinant vaccinia-CEA/TRICOM admixed with recombinant murine granulocyte macrophage colony-stimulating factor and then given low-dose interleukin 2. Mice were boosted on days 21, 28, and 35 with rF-CEA/TRICOM admixed with rF-granulocyte macrophage colony-stimulating factor and then given low-dose interleukin 2. Mice were monitored for survival and compared with groups of mice vaccinated in a similar manner with poxviral vectors containing CEA/B7-1 or CEA transgenes. To determine the mechanism of antitumor therapy, mice were depleted of T-cell subpopulations before vaccination with the CEA/TRICOM regimen. Mice successfully cured of tumor and age-matched control mice were monitored for 1 year. At 1 year, several clinical assays were carried out involving analysis of 9 serological parameters, 11 urinalysis parameters, and 14 immunological parameters. In addition, histopathology was performed on 42 tissues/mouse. RESULTS: The CEA/TRICOM vaccination regimen induced a therapeutic antitumor response as measured by increased survival, which was due largely to induced T cell responses (both CD4(+) and CD8(+)) as determined by selective T-cell subset depletion. The CEA/TRICOM vaccination regimen induced a significant increase in proliferation of CD4(+) T cells to CEA protein and a significant increase in secretion of IFN-gamma from CD8(+) T cells in response to a defined CEA epitope. Despite CEA expression in normal adult gastrointestinal tissues, no toxicity was observed in the CEA/TRICOM-vaccinated group when an array of clinical serum and urine chemistry assays was conducted 1 year after vaccination. Moreover, a comprehensive histopathological evaluation of all tissues from these groups also showed no evidence of toxicity. CONCLUSIONS: Activation of T cells directed against a tumor-associated self-antigen, sufficient to mediate therapeutic antitumor immunity, was observed in vivo without the development of autoimmunity as analyzed by a comprehensive evaluation of biochemical, immunological, and histopathological criteria. These studies demonstrate that the use of vectors containing as many as three costimulatory molecules does not induce autoimmunity or other pathology. These studies thus demonstrate that a balance can indeed be achieved between the induction of an immune response to a self-antigen, which is capable of antitumor therapy, and the absence of autoimmunity. PMID- 12738743 TI - Identification of HLA-A*0201-restricted cytotoxic T lymphocyte epitope from TRAG 3 antigen. AB - PURPOSE: Identification of tumor antigen and subsequent identification of T-cell epitope from these antigens make specific immunotherapy for malignant tumor applicable. Because TRAG-3 antigen is expressed in most melanomas and 54% of non small cell lung carcinomas and HLA-A2.1-expressing individuals cover >50% in the population of China, we aim at identifying TRAG-3-encoded peptide presented by HLA-A2.1. EXPERIMENTAL DESIGN: In our study, a HLA-A2.1-restricted CTL epitope was identified by using the following four-step procedure: (a) computer-based epitope prediction from the amino acid sequence of TRAG-3 antigen; (b) peptide binding assay to determine the affinity of the predicted peptide with HLA-A2.1 molecule; (c) stimulation of primary T-cell response against the predicted peptides in vitro; and (d) testing of the induced CTLs toward LB373-MEL cells expressing TRAG-3 antigen and HLA-A2.1. RESULTS: Of the four tested peptides, effectors induced by a peptide of TRAG-3 at residue position 58-66 lysed LB373 MEL cells expressing both TRAG-3 and HLA-A2.1. Our results indicate that peptide TRAG-3(58 approximately 66) (ILLRDAGLV) is a new HLA-A2.1-restricted CTL epitope capable of inducing TRAG-3 specific CTLs in vitro. CONCLUSIONS: Because TRAG-3 is a cancer/testis antigen expressed in most melanomas and half of non-small cell lung carcinomas, identification of the TRAG-3/HLA-A2.1 peptide ILLRDAGLV may facilitate peptide-based specific immunotherapy for various histological tumors. PMID- 12738744 TI - Administration of optimal biological dose and schedule of interferon alpha combined with gemcitabine induces apoptosis in tumor-associated endothelial cells and reduces growth of human pancreatic carcinoma implanted orthotopically in nude mice. AB - PURPOSE: We determined whether chronic administration of IFN-alpha at optimal biological dose inhibits angiogenesis of human pancreatic carcinoma growing in the pancreas of nude mice. EXPERIMENTAL DESIGN: Cells of the human pancreatic cancer cell line L3.6pl were implanted into the pancreas of nude mice. Seven days later, groups of mice received s.c. injection with IFN-alpha alone (50,000 units biweekly or 10,000 units daily), i.p. injection with gemcitabine alone (125 mg/kg biweekly), or injection with both daily IFN-alpha and biweekly gemcitabine for 35 days. In a survival study, the mice were treated until they became moribund. RESULTS: Biweekly treatments with 50,000 units of IFN-alpha alone were ineffective. In contrast, daily injections of IFN-alpha (10,000 units/day) alone, biweekly injections of gemcitabine alone, or the combination of IFN-alpha and gemcitabine reduced tumor volume by 53%, 70%, and 87%, respectively. Immunohistochemical analysis revealed that treatment with IFN-alpha alone or with IFN-alpha plus gemcitabine inhibited expression of the proangiogenic molecules basic fibroblast growth factor and matrix metalloproteinase 9 more than did treatment with gemcitabine alone. These treatments also decreased the staining of proliferating cell nuclear antigen within the tumor and induced apoptosis in tumor-associated mouse endothelial cells (staining with CD31/terminal deoxynucleotidyl transferase-mediated nick end labeling), leading to a decrease in microvessel density. CONCLUSIONS: These data show that administration of IFN alpha at optimal biological dose and schedule in combination with gemcitabine induced apoptosis in tumor-associated endothelial cells and decreased growth of human pancreatic cancer cells in the pancreas, leading to a significant increase in survival. PMID- 12738746 TI - Cyclin d1 overexpression sensitizes breast cancer cells to fenretinide. AB - PURPOSE: Fenretinide has shown promise in the chemoprevention of breast cancer, a tumor type in which the oncogene cyclin D1 is overexpressed frequently. We aimed at determining the effect of cyclin D1 level on the response to fenretinide treatment. EXPERIMENTAL DESIGN: Stable clones of T47-D cells were created to overexpress cyclin D1 or a mutant of cyclin D1, injected in nude mice for xenograft formation, and the rate of tumor growth and tumor regression determined. RESULTS: We show here that cells overexpressing cyclin D1 are significantly more sensitive to fenretinide than genetically matched cells that express low levels of cyclin D1, and that fenretinide prevents tumor formation arising from cyclin D1-overexpressing cells. Furthermore, we show that fenretinide is also able to promote the regression of cyclin D1-positive tumors. We also show that cells expressing a mutant of cyclin D1 that cannot bind to cdk4 are also more sensitive to fenretinide. CONCLUSIONS: These results suggest that fenretinide may be particularly useful in the treatment of cyclin D1-positive breast cancers, and that the interaction between cyclin D1 and fenretinide is independent of cyclin D1 binding to cdk4. PMID- 12738745 TI - Specific and high-level targeting of radiolabeled octreotide analogues to human medulloblastoma xenografts. AB - PURPOSE: The objective of this study was to determine the feasibility of exploiting the overexpression of somatostatin subtype-2 receptors (sstr(2)) on human medulloblastoma cells to develop targeted radiodiagnostics and radiotherapeutics for this disease. EXPERIMENTAL DESIGN: The following radioiodinated peptides were prepared using chloramine-T and evaluated: [(131)I Tyr(3)]octreotide ([(131)I]TOC), [(131)I-Tyr(3)]octreotate ([(131)I]TOCA), involving substitution of Thr(ol)(8) in TOC with Thr(8), and glucose-[(131)I Tyr(3)]octreotide ([(131)I]Gluc-TOC) and glucose-[(131)I-Tyr(3)]octreotate ([(131)I]Gluc-TOCA), prepared by conjugation of glucose to the peptide NH(2) terminus. Specific internalization of the peptides by sstr(2)-expressing AR42J rat pancreatic carcinoma cells in vitro was evaluated in paired-label assays. The tissue distribution of i.v. administered [(131)I]TOC, [(131)I]TOCA, [(131)I]Gluc TOC, and [(131)I]Gluc-TOCA was evaluated in athymic mice bearing s.c. D341 Med human medulloblastoma xenografts. RESULTS: Compared with [(125)I]TOC, internalized radioiodine levels were higher for the other three peptides. For example, internalized counts were 1.9 +/- 0.2, 2.0 +/- 0.3, and 5.7 +/- 1.9 times higher for [(131)I]Gluc-TOC, [(131)I]TOCA, and [(131)I]Gluc-TOCA after a 3-h incubation, respectively, demonstrating that carbohydration and COOH-terminus modification significantly improved the retention of radioiodine activity in sstr(2)-expressing tumor cells. COOH-terminus modification significantly increased (131)I localization in D341 Med medulloblastoma xenografts [[(131)I]TOCA, 8.1 +/- 2.2% of injected dose/g (% ID/g); [(131)I]TOC, 3.9 +/- 0.5% ID/g at 1 h], whereas carbohydration of the NH(2) terminus resulted in even higher gains in tumor accumulation ([(131)I]Gluc-TOC, 11.1 +/- 1.8% ID/g; [(131)I]Gluc-TOCA, 21.4 +/- 7.3% ID/g). In addition, the three modified peptides exhibited liver activity levels that were less than half those of [(131)I]TOC. Uptake of the two glucose-peptide conjugates in this human medulloblastoma xenograft was blocked by coinjection of 100 micro g of octreotide, demonstrating that it was receptor-specific. Tumor:normal tissue uptake ratios for [(131)I]Gluc TOCA generally were higher that those for [(131)I]Gluc-TOC. At 1 h, tumor:normal tissue ratios for [(131)I]Gluc-TOCA were 29:1, 15:1, 8:1, 8:1, 240:1, and 82:1 for blood, liver, kidney, spleen, brain, and muscle, respectively. CONCLUSIONS: Our findings suggest that additional investigation of radiolabeled Gluc-TOCA analogues for the imaging and targeted radiotherapy of medulloblastoma is warranted. PMID- 12738747 TI - The cyclooxygenase 2-selective inhibitor NS398 inhibits proliferation of oral carcinoma cell lines by mechanisms dependent and independent of reduced prostaglandin E2 synthesis. AB - PURPOSE: We investigated the potential of cyclooxygenase (COX)-2 as anappropriate chemopreventive and/or therapeutic target for oral cancer. EXPERIMENTAL DESIGN: Immunohistochemical analysis of COX-2 expression was carried out on 37 oral squamous cell carcinomas (OSCCs) and 23 normal oral epithelium samples. We investigated whether the COX-2-selective inhibitor NS398 induced growth inhibition in four human OSCC cell lines and whether this was COX-2 dependent. RESULTS: COX-2 staining was more intense in the carcinomas compared with normal epithelium (P < 0.001). Early-stage tumors (stages I and II) had significantly higher epithelial COX-2 staining than late-stage tumors (stages III and IV; P = 0.034), and overexpression of COX-2 was detected in hyperplastic and dysplastic epithelium. Treatment of OSCC cells with NS398 for 72 h at concentrations of 50 micro M and above resulted in growth inhibition accompanied by a reversible G(0) G(1) arrest, but no apoptosis or terminal differentiation. However, a concentration of 10 micro M was sufficient to abolish secreted prostaglandin E(2) (PGE(2)) production. Over a longer treatment time, lower concentrations of NS398 were growth inhibitory. Growth inhibition of the OSCC cell line H357 was detected after treatment with 5 micro M NS398 as well as 100 micro M NS398 for 6-12 days. In cultures treated with 5 micro M NS398, but not in those treated with 100 micro M NS398, restoration of PGE(2) to control levels abrogated growth inhibition. CONCLUSIONS: NS398 inhibits the growth of OSCC cells by mechanisms that are dependent and independent of suppression of PGE(2) synthesis. Molecular targeting of COX-2, PGE(2) synthase, or PGE(2) receptors may be useful as a chemopreventive or therapeutic strategy for oral cancer. PMID- 12738748 TI - Dual Action of NAMI-A in inhibition of solid tumor metastasis: selective targeting of metastatic cells and binding to collagen. AB - NAMI-A is a ruthenium complex endowed with a selective effect on lung metastases of solid metastasizing tumors. The aim of this study is to provide evidence that NAMI-A's effect is based on the selective sensitivity of the metastasis cell, as compared with other tumor cells, and to show that lungs represent a privileged site for the antimetastatic effects. The transplantation of Lewis lung carcinoma cells, harvested from the primary tumor of mice treated with 35 mg/kg/day NAMI-A for six consecutive days, a dose active on metastases, shows no change in primary tumor take and growth but a significant reduction in formation of spontaneous lung metastases. Transmission electron microscopy examination of lungs and kidney shows NAMI-A to selectively bind collagen of the lung extracellular matrix and also type IV collagen of the basement membrane of kidney glomeruli. The half lifetime of NAMI-A elimination from the lungs is longer than for liver, kidney, and primary tumor. NAMI-A bound to collagen is active on tumor cells as shown in vitro by an invasion test, using a modified Boyden chamber and Matrigel, and it inhibits the matrix metallo-proteinases MMP-2 and MMP-9 at micromolar concentrations, as shown in vitro by a zimography test. These data show NAMI-A to significantly affect tumor cells with metastatic ability. Binding to collagen allows NAMI-A to exert its selective activity on metastatic cells during dissemination and particularly in the lungs. These data also stress the wide spectrum of daily doses and treatment schedules at which NAMI-A is active against metastases. PMID- 12738749 TI - Novel kidney cancer immunotherapy based on the granulocyte-macrophage colony stimulating factor and carbonic anhydrase IX fusion gene. AB - PURPOSE: We investigated the ability of the fusion protein granulocyte-macrophage colony-stimulating factor and carbonic anhydrase IX (GMCA-9)(1) to induce an immune response in vitro and in vivo for the development of a GMCA-9-based kidney cancer vaccine. EXPERIMENTAL DESIGN: Human dendritic cells (DCs) were transduced with a recombinant adenovirus containing the GMCA-9 gene and tested for their capacity to induce CA9-specific cytotoxic T lymphocytes in vitro. Tumor growth was studied in severe compromised immunodeficiency disease (SCID) mice s.c. injected with R11-GMCA-9, a human renal cell carcinoma cell line stably transfected with the GMCA-9 gene. Involvement of natural killer (NK) cells in the antitumor activity of GMCA-9 was determined in SCID mice treated with the NK blocking agent anti-asialoGM-1. RESULTS: DC and R11 cells transduced with GMCA-9 produced a GMCA-9 protein that is targeted to the cell membrane and partially processed to granulocyte macrophage colony-stimulating factor- and CA9-like products. Furthermore, GMCA-9 was capable of inducing DC maturation, as well as CA9-specific cytotoxic lymphocytes in vitro. Tumor growth of R11 cells in SCID mice was significantly inhibited after transfection with the GMCA-9 fusion gene (P < 0.01). In mice treated with anti-asialoGM-1, R11-GMCA-9 tumors grew significantly faster than those of control mice (P < 0.05), suggesting an involvement of NK cells. CONCLUSIONS: Our results suggest that the fusion protein GMCA-9 is capable of generating an immune response both in vitro and in vivo. Additional studies will confirm the utility of ex vivo GMCA-9-transduced DCs as a kidney cancer vaccine. PMID- 12738750 TI - In vitro and in vivo antitumor activity of methotrexate conjugated to human serum albumin in human cancer cells. AB - To avoid systemic toxicity of the cytotoxic drug methotrexate (MTX) and to improve tumor selectivity, MTX was bound to human serum albumin (HSA) as a drug carrier. To understand more about the mechanism of action of MTX conjugated to HSA (MTX-HSA), the uptake of MTX-HSA into the cell was determined as well as the effect of MTX-HSA on thymidylate synthase (TS), cell cycle distribution, and cell proliferation. Different uptake kinetics were observed for [(3)H]MTX and [(3)H]MTX-HSA. However, similar uptake kinetics were measured for (125)I-HSA and (125)I-MTX-HSA (2.1 and 1.8 pmol/10(7) cells/h when cells were treated with 10 micro M (125)I-HSA and (125)I-MTX-HSA, respectively), suggesting that MTX-HSA enters the cells by albumin-mediated endocytosis. We observed no effect of MTX HSA on TS when folate receptor-expressing KB cells were treated for 4 h (IC(50), >50 micro M). However, 24 h after incubation, MTX-HSA inhibited TS with an IC(50) of 6.9 micro M. In addition, we found that MTX-HSA had a delayed effect on the cell cycle compared with MTX and that this effect could be inhibited with the lysosomal inhibitor methylamine, suggesting that MTX-HSA activity is dependent on lysosomal processes. The proliferation of different wild-type and MTX-resistant tumor cell lines was inhibited at IC(50) concentrations between 2 and 78 micro M, respectively. MTX-HSA accumulates in vivo in the tumor tissue. Local concentrations of 18-29 micro M were measured, which are effective antiproliferative concentrations as determined in vitro. We also investigated the antitumor activity of MTX-HSA in vivo in different human tumor xenografts grown s.c. in nude mice. Fourteen tumors from eight different tissues were tested. Nine of 14 tumors (64%) showed a clear response with tumor inhibition, stasis, or regression; 5 of 14 (36%) gave a moderate response with tumor growth delay or no response. In conclusion, MTX-HSA is effectively taken up by the cells via albumin receptor- or folate receptor-mediated endocytosis and time-dependently released as an active compound into the cytosol to exert an inhibiting effect on TS and to induce cell cycle alterations. In vivo, effective concentrations of MTX-HSA were reached in tumor tissue to exhibit antitumor activity. PMID- 12738751 TI - Chemoprevention of benzo(a)pyrene-induced lung tumors in mice by the farnesyltransferase inhibitor R115777. AB - PURPOSE: Inhibitors of farnesyltransferase (e.g., R115777) are being developed for therapy and prevention of various cancers. The efficacy of R115777 [Zarnestra; (B)-6-[amino(4-chlorophenyl)(1-methyl-1H-imidazol-5-yl)-methyl]-4-(3 chlorophenyl)-1-methyl-2(1H)-quinolinone] to prevent the development of lung tumors in mice was determined. EXPERIMENTAL DESIGN: Female strain A mice (7-8 weeks of age) were given 100 mg/kg benzo(a)pyrene [B(a)P] by i.p. injection, and 4 or 14 weeks later, they were given 50 or 100 mg/kg R115777 by oral gavage 5 days/week. The mice were sacrificed 22 weeks after they received the B(a)P. RESULTS: Tumor multiplicity was 5.0 +/- 0.85, 4.5 +/- 0.52, 2.1 +/- 0.31, and 1.5 +/- 0.31 tumors/mouse in mice that received 0, 50, 100 (weeks 4-22), or 100 (weeks 14-22) mg/kg R115777. Thus, 100 mg/kg R115777 was similarly effective in preventing lung tumors when administered during the promotional phase of carcinogenesis [that is, either 4 or 14 weeks after B(a)P], whereas the lower dose of 50 mg/kg R115777 was ineffective. The proliferating cell nuclear antigen labeling index was also significantly reduced in lung tumors from mice treated with 100 mg/kg R115777 starting at 4 or 14 weeks. CONCLUSIONS: These results demonstrated that R115777 can prevent the development of lung tumors in the A/J mouse model, where tumors routinely have mutations in the Ki-Rasoncogene. PMID- 12738752 TI - Bcl-2 antisense (G3139, Genasense) enhances the in vitro and in vivo response of Epstein-Barr virus-associated lymphoproliferative disease to rituximab. AB - Bcl-2 is up-regulated by EBV in immortalized lymphoblastoid B cells and is expressed in the majority of EBV-associated lymphoproliferative diseases, including posttransplant lymphoproliferative disorder (PTLD) and AIDS-related lymphoma (ARL). Given the antiapoptotic and chemoprotective effect of Bcl-2, it represents a logical target for modulation using antisense strategies in PTLD and ARL. We previously examined the antitumor effects of a fully phosphorothioated Bcl-2 antisense oligonucleotide, G3139, in EBV(+) lymphoproliferative disease in vitro and in vivo using the human/severe combined immunodeficient (SCID) chimeric model of PTLD. These studies showed that G3139 treatment decreased Bcl-2 protein levels in association with antiproliferative and proapoptotic effects in lymphoblastoid cell lines (LCLs) in vitro. In vivo, although G3139 treatment completely abrogated EBV(+) lymphoid tumor engraftment in the human/SCID model of PTLD, antisense treatment alone was not curative in animals with established tumors. Because the humanized anti-CD20 antibody, rituximab, has antitumor activity in patients with PTLD and stimulates apoptosis in some lymphoid cell lines, we sought to determine whether Bcl-2 antisense treatment potentiates the antitumor effects of rituximab in EBV-associated lymphoproliferative disease in vitro and in vivo. Proliferation assays by thymidine uptake in LCLs showed that G3139 but not control oligonucleotides augmented the antiproliferative effect of rituximab. Flow cytometric terminal deoxynucleotidyltransferase-mediated nick end labeling assays confirmed that G3139 treatment enhanced the apoptotic response of LCLs to rituximab, and this interaction was oligonucleotide sequence dependent. To test the in vivo efficacy of G3139 and rituximab in the human/SCID model of PTLD, we used a delayed treatment schedule that permitted detection of enhanced antitumor activity of combination therapy. Although G3139 or rituximab treatment significantly prolonged survival compared with untreated controls, 89% of animals in the monotherapy arms died with disseminated tumors. In contrast, 79% of animals in the combined G3139 and rituximab arm remained tumor free for the duration of follow-up (>160 days) with no evidence of tumors at the time of sacrifice, indicating that G3139 in combination with rituximab was curative therapy in the majority of tumor-bearing animals. These studies demonstrated that G3139 potentiates the antitumor response of PTLD to rituximab in vivo and augments the antiproliferative and apoptotic effects of rituximab in vitro in LCLs. This is the first report of G3139 potentiating the antitumor activity of an antibody-based therapy both in vitro and in vivo. Bcl-2 antisense oligonucleotide therapy in combination with rituximab may represent a promising nontoxic and effective targeted therapy for EBV-associated lymphoproliferative diseases such as PTLD and ARL. Furthermore, this approach may have broader applications to other Bcl-2- and CD20-expressing lymphoid malignancies. PMID- 12738755 TI - Foreword. PMID- 12738753 TI - Biochemical and genetic association of plasma apolipoprotein A-II levels with familial combined hyperlipidemia. AB - Apolipoprotein A-II (apoA-II) is a major protein on high-density lipoprotein (HDL) particles, and in mice, its levels are associated with triglyceride and glucose metabolism. In particular, transgenic mice overexpressing apoA-II exhibit hypertriglyceridemia, increased body fat, and insulin resistance, whereas apoA-II null mice have decreased triglycerides and increased insulin sensitivity. Given the phenotypic overlap between familial combined hyperlipidemia (FCH) and apoA-II transgenic mice, we investigated the relationship of apoA-II to this disorder. Despite having lower HDL-cholesterol (HDL-C), FCH subjects had higher apoA-II levels compared with unaffected relatives (P<0.00016). Triglyceride and HDL-C levels were significant predictors of apoA-II, demonstrating that apoA-II variation is associated with several FCH-related traits. After adjustment for multiple covariates, there was evidence for the heritability of apoA-II levels (h2=0.15; P<0.02) in this sample. A genome scan for apoA-II levels identified significant evidence (LOD=3.1) for linkage to a locus on chromosome 1q41, coincident with a suggestive linkage for triglycerides (LOD score=1.4). Thus, this locus may have pleiotropic effects on apoA-II and FCH traits. Our results demonstrate that apoA-II is biochemically and genetically associated with FCH and may serve as a useful marker for understanding the mechanism by which FCH develops. PMID- 12738754 TI - Hypercapnic acidosis activates KATP channels in vascular smooth muscles. AB - ATP-sensitive K+ channels (KATP) couple intermediary metabolism to cellular activity, and may play a role in the autoregulation of vascular tones. Such a regulation requires cellular mechanisms for sensing O2, CO2, and pH. Our recent studies have shown that the pancreatic KATP isoform (Kir6.2/SUR1) is regulated by CO2/pH. To identify the vascular KATP isoform(s) and elucidate its response to hypercapnic acidosis, we performed these studies on vascular smooth myocytes (VSMs). Whole-cell and single-channel currents were studied on VSMs acutely dissociated from mesenteric arteries and HEK293 cells expressing Kir6.1/SUR2B. Hypercapnic acidosis activated an inward rectifier current that was K+-selective and sensitive to levcromakalim and glibenclamide with unitary conductance of approximately 35pS. The maximal activation occurred at pH 6.5 to 6.8, and the current was inhibited at pH 6.2 to 5.9. The cloned Kir6.1/SUR2B channel responded to hypercapnia and intracellular acidification in an almost identical pattern to the VSM current. In situ hybridization histochemistry revealed expression of Kir6.1/SUR2B mRNAs in mesenteric arteries. Hypercapnia produced vasodilation of the isolated and perfused mesenteric arteries. Pharmacological interference of the KATP channels greatly eliminated the hypercapnic vasodilation. These results thus indicate that the Kir6.1/SUR2B channel is a critical player in the regulation of vascular tones during hypercapnic acidosis. PMID- 12738756 TI - Renal function as a predictor of long-term graft survival in renal transplant patients. AB - Acute rejection is a major risk factor for kidney graft failure. However, as acute rejection has been progressively reduced by recent immunosuppressive regimens, other risk factors are becoming increasingly important. Evidence is accumulating that early renal function predicts long-term outcome. A recent registry survey of more than 100 000 kidney transplants found that 6- and 12 month serum creatinine levels, as well as the change between 6 and 12 months, are strongly associated with long-term graft survival. A survey of paediatric renal transplant recipients showed that poor creatinine clearance (<50 ml/min) as early as 30 days post-transplant predicted an annual rate of graft loss of 13% compared with <3% in patients with 30-day clearance >50 ml/min. This association between early renal function and long-term outcome was confirmed in multicentre studies. Renal transplant recipients (n=572) with 6-month serum creatinine levels >1.5 mg/dl suffered 3-year graft loss of 19.3% compared with only 8.5% in patients with levels <1.6 mg/dl (P<0.001). Significantly fewer patients receiving tacrolimus had 12-month serum creatinine levels >1.5 mg/dl compared with cyclosporin (42 versus 54%, P<0.05). Interestingly, a single-centre study (n=436) found that while glomerular filtration rate (GFR) at 6 months post-transplant had remained stable over the last decade, the rate of loss of renal function had decreased. A lower rate of GFR loss was associated with absence of rejection, use of mycophenolate mofetil rather than azathioprine and use of tacrolimus rather than cyclosporin (P<0.01). In conclusion, early measures of renal function allow identification of those patients at highest risk of graft failure and provide an invaluable tool for improving outcomes by tailored immunosuppression. The choice of such immunosuppression should be guided not only by its ability to prevent rejection, but also by its impact on renal function. PMID- 12738757 TI - Tacrolimus versus cyclosporin immunosuppression: long-term outcome in renal transplantation. AB - Despite reduced risk of acute rejection and increased 1-year graft survival with modern immunosuppressive regimens, chronic allograft nephropathy and death with a functioning graft remain major causes of allograft loss beyond the first year post-transplant. Anti-rejection agents may influence renal transplant outcome not solely through their immunosuppressive activity but also through their effects on other prognostic risk factors. We have analysed 6-year follow-up data from 232 renal transplant recipients randomized to treatment with tacrolimus or cyclosporin microemulsion at the University Hospital of Wales, Cardiff. Tacrolimus-based therapy was associated with a more favourable cardiovascular risk profile than therapy with cyclosporin microemulsion, with an improved lipid profile, lower arterial blood pressure and lower homocysteine levels. Renal function at 1-year post-transplant is probably the most significant factor influencing long-term graft survival. In our analyses, renal function determined by the glomerular filtration rate was significantly better in tacrolimus-treated patients from month 3 post-transplant. Moreover, normal renal function was maintained throughout a 5-year follow-up in a significantly higher proportion of non-rejecting patients treated with tacrolimus than with cyclosporin microemulsion (58 versus 10%, respectively, at 5 years; P=0.002). Morphometric analysis of protocol biopsies revealed that the degree of interstitial fibrosis, similar in both treatment groups at baseline, was significantly greater in the cyclosporin microemulsion group over 12 months. Importantly, patients receiving tacrolimus had significantly greater 6-year graft survival (81 versus 60%, P=0.0496) and a higher projected graft half-life (15 versus 10 years) than those receiving cyclosporin microemulsion. Tacrolimus treatment is associated with a significantly better cardiovascular risk profile and superior renal function compared with cyclosporin microemulsion treatment, which appears to translate into improved long-term graft survival. PMID- 12738758 TI - Pharmacokinetics of tacrolimus-based combination therapies. AB - This paper reviews the pharmacokinetics of tacrolimus, with special reference to its combination with adjunctive immunosuppressants. Oral bioavailability of tacrolimus, which is variable between patients, averages approximately 25%. This is largely due to extrahepatic metabolism of tacrolimus in the gastrointestinal epithelium. Nevertheless, intra-patient variability is low, as evidenced by the small number of dose changes required to maintain patients within the recommended tacrolimus target levels. Tacrolimus is distributed extensively in the body with most partitioned outside the blood compartment. Concentrations of tacrolimus in blood are used as a surrogate marker of clinically relevant concentration of the drug at the site(s) of action. Convenient whole-blood sampling within a +/-2-h window around 12 h post-dose (C(min)) is highly predictive of systemic exposure to tacrolimus and is thus used to optimise therapy. Sampling at other time-points offers no advantage over C(min) monitoring. The interactions of tacrolimus with other immunosuppressive agents are well characterized. After cessation of concomitant corticosteroid treatment, exposure to tacrolimus increases by approximately 25%. In contrast, there is no pharmacokinetic interaction between mycophenolate mofetil (MMF) and tacrolimus. Therefore, systemic exposure to the active metabolite of MMF, mycophenolic acid, is higher with MMF-tacrolimus combination than with MMF-cyclosporin combination. Therefore, 1 g/day MMF may be an adequate maintenance dose in tacrolimus-based regimens. Co-administration of tacrolimus and sirolimus, while having no effect on exposure to sirolimus, results in reduced exposure to tacrolimus at sirolimus doses of 2 mg/day and above. In conclusion, tacrolimus levels should be monitored when sirolimus is co administered at doses >2 mg/day and after cessation of corticosteroid treatment. PMID- 12738759 TI - Tailoring tacrolimus-based immunotherapy in renal transplantation. AB - Tacrolimus is a cornerstone immunosuppressive agent in renal transplantation and compared with cyclosporin, its use is associated with a reduced incidence of acute rejection. Optimizing immunosuppressive management in the early post transplant period is important for achieving long-term graft function and survival. In attempts to improve the long-term outcomes of renal transplantation further, tacrolimus has been combined with two novel immunosuppressive agents, mycophenolate mofetil (MMF) and sirolimus, with encouraging results in terms of patient and graft survival, acute rejection rates and renal graft function. Tacrolimus in combination with MMF adjunctive therapy showed significantly better graft survival in patients with delayed graft function, fewer episodes of corticosteroid-resistant rejection and better renal function at the 3-year follow up compared with cyclosporin microemulsion plus MMF immunosuppression. A tacrolimus plus MMF regimen was also effective for renal transplant recipients at our centre in Pennsylvania, resulting in excellent survival and rejection rates at 1 year post-transplantation. The 3-month results of a US multicentre study comparing tacrolimus in combination with either MMF or sirolimus showed these two treatment regimens to be equivalent in terms of patient and graft survival, delayed graft function, the incidence of biopsy-confirmed acute rejection and renal graft function, although differences were apparent in terms of acute tubular necrosis and hyperlipidaemia. In conclusion, the development of a new immunosuppressive regimen in renal transplantation should take account of factors that influence graft function, both in the short and long term, as a way of optimizing individual maintenance therapy. PMID- 12738760 TI - Role of pleckstrin homology domain in regulating membrane targeting and metabolic function of insulin receptor substrate 3. AB - The insulin receptor phosphorylates insulin receptor substrate (IRS) proteins on multiple tyrosine residues that act as docking sites to recruit a number of downstream signaling molecules. Here we show that IRS3 is localized both at the plasma membrane and in the nucleus. Interestingly, the nuclear localization of the protein is restricted to specific regions involved in mRNA processing and known as speckles. By using different truncated versions of the protein, we demonstrate that the pleckstrin homology (PH) domain is involved in IRS3 localization at the level of both plasma membrane and nucleus. To our knowledge this is the first report of a PH domain responsible for a nuclear targeting of the host protein. By site-directed mutagenesis, we identify residues within the PH domain critical for proper localization of IRS3. Mutations within the PH domain preventing IRS3 intracellular localization result in an inhibition of IRS3 induced glucose uptake. We conclude that the PH domain is required for IRS3 intracellular localization and, furthermore, that it has a key role in metabolic functions of IRS3. In particular, our data suggest that IRS3 intracellular localization at the plasma membrane and in the nucleus is the result of two different cooperative mechanisms both involving the PH domain. PMID- 12738761 TI - Transactivation of the epidermal growth factor receptor mediates parathyroid hormone and prostaglandin F2 alpha-stimulated mitogen-activated protein kinase activation in cultured transgenic murine osteoblasts. AB - Recent data suggest that G protein-coupled receptors (GPCRs), including those for PTH and prostaglandins (PGs), contribute to the proliferation and differentiation of osteoblasts in vivo. To understand how these signals are transduced, we studied activation of the ERK1/2 MAPK cascade in cultures of differentiating TMOb murine osteoblasts. In TMOb cells, stimulation of endogenous Gs/Gq-coupled PTH receptors, Gq-coupled PGF2 alpha receptors, and Gi/Gq-coupled lysophosphatidic acid receptors, but not Gs-coupled PGE2 receptors, caused a rapid 5- to 10-fold increase in ERK1/2 phosphorylation. GPCR-stimulated ERK1/2 activation coincided with increased tyrosine phosphorylation of epidermal growth factor (EGF) receptors and was blocked by the EGF receptor inhibitor, tyrphostin AG1478, and the metalloprotease inhibitor, batimastat, suggesting that the response involved transactivation of EGF receptors through the proteolytic release of an EGF receptor ligand. To further examine the mechanism of PTH-stimulated EGF receptor transactivation, we employed COS-7 cells expressing the rat PTH receptor. Here, stimulation with PTH(1-34) caused proteolysis of hemagglutinin epitope-tagged heparin binding-EGF, increased tyrosine autophosphorylation of EGF receptors, and AG1478-sensitive ERK1/2 activation. When PTH receptor-expressing COS-7 cells were placed in a mixed culture with cells lacking the PTH receptor but expressing a green fluorescent protein-tagged ERK2, stimulation with PTH(1-34) induced phosphorylation of green fluorescent protein-ERK2 that was abolished by either batimastat or tyrphostin AG1478. These data suggest that autocrine/paracrine cross-talk between EGF receptors and Gi- or Gq/11-coupled GPCRs represents the predominant mechanism of GPCR-mediated activation of ERK1/2 in cultured TMOb osteoblasts. PMID- 12738762 TI - Fer is a downstream effector of insulin and mediates the activation of signal transducer and activator of transcription 3 in myogenic cells. AB - Fer is an intracellular tyrosine kinase that associates with signal transducer and activator of transcription 3 (Stat3) in mammalian cells. However, the signaling pathways in which this interaction plays a functional role have not been revealed. Herein, we show that insulin up-regulates the levels of the fer mRNA and Fer protein in myoblasts that undergo insulin-induced myogenic differentiation. Moreover, insulin directs the interaction of Fer with members of the Janus family of tyrosine kinases (Jak)-Stat3 signaling pathway. Although in untreated cells Fer binds Jak1 and its tyrosine phosphorylation level is low, insulin treatment induced the phosphorylation of Fer and its dissociation from Jak1. The up-regulation of Fer and its dissociation from Jak1 were accompanied by an augmented association of activated Fer with Stat3 and by a concomitant increase in the tyrosine phosphorylation of Stat3. Dissociation of Fer and Jak1, as well as elevation in the level of Fer and in the tyrosine phosphorylation of Stat3, depended on the activity of phosphatidylinositol 3-kinase (PI3K) and was abolished by a PI3K inhibitor. However, Fer and Stat3 were only mildly affected by low concentrations of IGF-I, another activator of the PI3K pathway that can also induce myogenic differentiation. RNA interference directed toward the fer mRNA did not affect the cellular levels of Stat3 but led to a dramatic reduction in the tyrosine phosphorylation level of this transcription factor. Thus, Fer is a downstream effector of insulin and mediates the activation of Stat3 in myogenic cells. PMID- 12738764 TI - Computer-assisted generation of a protein-interaction database for nuclear receptors. AB - With the increasing amount of biological data available, automated methods for information retrieval become necessary. We employed computer-assisted text mining to retrieve all protein-protein interactions for nuclear receptors from MEDLINE in a systematic way. A dictionary of protein names and of terms denoting interactions was generated, and trioccurrences of two protein names and one interaction term in one sentence were retrieved. Abstracts containing at least one such trioccurrence were manually checked by biologists to select the relevant interactions out of the automatically extracted data. In total, 4360 abstracts were retrieved containing data on protein interactions for nuclear receptors. The resulting database contains all reported protein interactions involving nuclear receptors from 1966 to September 2001. Remarkably, the annual increase in number of reported interactors for nuclear receptors has been following an exponential growth curve in the years 1991 to 2001. Apparent in the data set is the high complexity of protein interactions for nuclear receptors. The number of interactions correlates with the number of published papers for a given receptor, suggesting that the number of reported interactors is a reflection of the intensity of research dedicated to a given receptor. Indeed, comparison of the retrieved data to a systematic yeast two-hybrid-based interaction analysis suggests that most NRs are similar with respect to the number of interacting proteins. The data set obtained serves as a source for information on NR interactions, as well as a reference data set for the improvement of advanced text-mining methods. PMID- 12738763 TI - RET/PTC (rearranged in transformation/papillary thyroid carcinomas) tyrosine kinase phosphorylates and activates phosphoinositide-dependent kinase 1 (PDK1): an alternative phosphatidylinositol 3-kinase-independent pathway to activate PDK1. AB - Thyroid cancers are a leading cause of death due to endocrine malignancies. RET/PTC (rearranged in transformation/papillary thyroid carcinomas) gene rearrangements are the most frequent genetic alterations identified in papillary thyroid carcinoma. Although the oncogenic potential of RET/PTC is related to intrinsic tyrosine kinase activity, the substrates for this enzyme are yet to be identified. In this report, we show that phosphoinositide-dependent kinase 1 (PDK1), a pivotal serine/threonine kinase in growth factor-signaling pathways, is a target of RET/PTC. RET/PTC and PDK1 colocalize in the cytoplasm. RET/PTC phosphorylates a specific tyrosine (Y9) residue located in the N-terminal region of PDK1. Y9 phosphorylation of PDK1 by RET/PTC requires an intact catalytic kinase domain. The short (iso 9) and long forms (iso 51) of the RET/PTC kinases (RET/PTC1 and RET/PTC3) induce Y9 phosphorylation of PDK1. Moreover, Y9 phosphorylation of PDK1 by RET/PTC does not require phosphatidylinositol 3-kinase or Src activity. RET/PTC-induced phosphorylation of the Y9 residue results in increased PDK1 activity, decrease of cellular p53 levels, and repression of p53 dependent transactivation. In conclusion, RET/PTC-induced tyrosine phosphorylation of PDK1 may be one of the mechanisms by which it acts as an oncogenic tyrosine kinase in thyroid carcinogenesis. PMID- 12738765 TI - Crystal structures of RbsD leading to the identification of cytoplasmic sugar binding proteins with a novel folding architecture. AB - RbsD is the only protein whose biochemical function is unknown among the six gene products of the rbs operon involved in the active transport of ribose. FucU, a paralogue of RbsD conserved from bacteria to human, is also the only protein whose function is unknown among the seven gene products of the l-fucose regulon. Here we report the crystal structures of Bacillus subtilis RbsD, which reveals a novel decameric toroidal assembly of the protein. Nuclear magnetic resonance and other studies on RbsD reveal that the intersubunit cleft of the protein binds specific forms of d-ribose, but it does not have an enzyme activity toward the sugar. Likewise, FucU binds l-fucose but lacks an enzyme activity toward this sugar. We conclude that RbsD and FucU are cytoplasmic sugar-binding proteins, a novel class of proteins whose functional role may lie in helping influx of the sugar substrates. PMID- 12738766 TI - Effect of protein kinase A-mediated phosphorylation on the structure and association properties of the enteropathogenic Escherichia coli Tir virulence protein. AB - Enteropathogenic Escherichia coli virulence is dependent on delivery of the translocated intimin receptor protein (Tir) into host cells. Tir phosphorylation on a single tyrosine (Tyr-474) is essential in mediating cytoskeletal rearrangement correlated with disease. Tir is also phosphorylated on other residues, with cAMP-dependent kinase (PKA) modification shown to play a role in Tir function. However, the mechanism by which nontyrosine phosphorylation affects Tir function remains unclear. In this study, analytical ultracentrifugation, SDS and native gel electrophoresis revealed that both Tir and its C-terminal domain (residues 385-550 of Tir that include the PKA substrate sites) exist in an equilibrium of monomers, dimers, and in the case of Tir, higher oligomers. PKA phosphorylation (1:300, PKA/C-Tir, mol/mol) shifted the equilibrium of C-Tir, but not Tir, predominantly to the dimeric state, whereas, at 100-fold higher concentrations of PKA the phosphorylated C-Tir was largely monomeric. This monomeric state was also produced at the lower PKA concentration and physiological ionic strength. Phosphorylation-mediated effects were achieved without significant changes in secondary structure as determined by circular dichroism spectroscopy. The data presented indicate that PKA-mediated phosphorylation induces changes in the association properties of the C-terminal domain of Tir that may facilitate Tir-Tir interactions and subsequently C-Tir host protein interactions in vivo. PMID- 12738767 TI - Interferon regulatory factor-2 regulates cell growth through its acetylation. AB - We have previously shown that interferon regulatory factor-2 (IRF-2) is acetylated by p300 and PCAF in vivo and in vitro. In this study we identified, by mass spectrometry, two lysine residues in the DNA binding domain (DBD), Lys-75 and Lys-78, to be the major acetylation sites in IRF-2. Although acetylation of IRF-2 did not alter DNA binding activity in vitro, mutation of Lys-75 diminished the IRF-2-dependent activation of histone H4 promoter activity. Acetylation of IRF-2 and IRF-2-stimulated H4 promoter activity were inhibited by the adenovirus E1A, indicating the involvement of p300/CBP. Mutation of Lys-78, a residue conserved throughout the IRF family members, led to the abrogation of DNA binding activity independently of acetylation. H4 is transcribed only in rapidly growing cells and its promoter activity is dependent on cell growth. Consistent with a role for acetylated IRF-2 in cell growth control, IRF-2 was acetylated only in growing NIH 3T3 cells, but not in growth-arrested counterparts. Chromatin immunoprecipitation assays showed that IRF-2 interacted with p300 and bound to the endogenous H4 promoter only in growing cells, although the levels of total IRF-2 were comparable in both growing and growth-arrested cells. These results indicate that IRF-2 is acetylated in a cell growth-dependent manner, which enables it to contribute to transcription of cell growth-regulated promoters. PMID- 12738768 TI - Mcm7, a subunit of the presumptive MCM helicase, modulates its own expression in conjunction with Mcm1. AB - The Saccharomyces cerevisiae Mcm7 protein is a subunit of the presumed heteromeric MCM helicase that melts origin DNA and unwinds replication forks. Previous work showed that Mcm1 binds constitutively to the MCM7 promoter and regulates MCM7 expression. Here, we identify Mcm7 as a novel cofactor of Mcm1 in the regulation of MCM7 expression. Transcription of MCM7 is increased in the mcm7 1 mutant and decreased in the mcm1-1 mutant, suggesting that Mcm7 modulates its own expression in conjunction with Mcm1. Indeed, Mcm7 stimulates Mcm1 binding to the early cell cycle box upstream of the promoters of MCM7 as well as CDC6 and MCM5. Whereas Mcm1 binds these promoters constitutively, Mcm7 is recruited during late M phase, consistent with Mcm7 playing a direct role in modulating the periodic expression of early cell cycle genes. The multiple roles of Mcm7 in replication initiation, replication elongation, and autoregulation parallel those of the oncoprotein, the large T-antigen of the SV40 virus. PMID- 12738769 TI - Activation of the c-Jun N-terminal kinase signaling cascade mediates the effect of amyloid-beta on long term potentiation and cell death in hippocampus: a role for interleukin-1beta? AB - Amyloid-beta (Abeta) is a major constituent of the neuritic plaque found in the brain of Alzheimer's disease patients, and a great deal of evidence suggests that the neuronal loss that is associated with the disease is a consequence of the actions of Abeta. In the past few years, it has become apparent that activation of c-Jun N-terminal kinase (JNK) mediates some of the effects of Abeta on cultured cells; in particular, the evidence suggests that Abeta-triggered JNK activation leads to cell death. In this study, we investigated the effect of intracerebroventricular injection of Abeta(1-40) on signaling events in the hippocampus and on long term potentiation in Schaffer collateral CA1 pyramidal cell synapses in vivo. We report that Abeta(1-40) induced activation of JNK in CA1 and that this was coupled with expression of the proapoptotic protein, Bax, cytosolic cytochrome c, poly-(ADP-ribose) polymerase cleavage, and Fas ligand expression in the hippocampus. These data indicate that Abeta(1-40) inhibited expression of long term potentiation, and this effect was abrogated by administration of the JNK inhibitor peptide, D-JNKI1. In parallel with these findings, we observed that Abeta-induced changes in caspase-3 activation and TdT mediated dUTP nick-end labeling staining in neuronal cultured cells were inhibited by D-JNKI1. We present evidence suggesting that interleukin (IL)-1beta plays a significant role in mediating the effects of Abeta(1-40) because Abeta(1 40) increased hippocampal IL-1beta and because several effects of Abeta(1-40) were inhibited by the caspase-1 inhibitor Ac-YVAD-CMK. On the basis of our findings, we propose that Abeta-induced changes in hippocampal plasticity are likely to be dependent upon IL-1beta-triggered activation of JNK. PMID- 12738770 TI - E3 ubiquitin ligase Smurf1 mediates core-binding factor alpha1/Runx2 degradation and plays a specific role in osteoblast differentiation. AB - Osteoblast differentiation and bone formation is stimulated by bone morphogenetic protein (BMP)-2 and its downstream signaling molecules Smad1 and -5 and the osteoblast-specific transcription factor core-binding factor alpha1 (Cbfa1). Proteolytic degradation of Smad1 and Cbfa1 is proteasome-dependent, and intracellular concentrations of Smad1 and Cbfa1 are enhanced by inhibition of the 26 S proteasome. Smad1 degradation is mediated by the E3 ubiquitin ligase Smurf1 (Smad ubiquitin regulatory factor 1), but the specific E3 ligase responsible for Cbfa1 degradation has not been identified. Because Cbfa1 interacts with Smad1, whose degradation is mediated by Smurf1, we examined the effect of Smurf1 on Cbfa1 degradation in osteoblast precursor cells. Smurf1 interacts directly with Cbfa1 and mediates Cbfa1 degradation in a ubiquitin- and proteasome-dependent manner. Because Smurf1 controls the intracellular concentrations of several key molecules in the bone formation cascade, we examined the effect of a mutant form of Smurf1 in osteoblasts and found that expression of mutant Smurf1 markedly enhanced osteoblast differentiation. Smurf1 therefore appears to be an important regulatory factor in osteoblast differentiation and a potential molecular target for identification of bone anabolic agents. PMID- 12738771 TI - Activation of the ATR-mediated DNA damage response by the HIV-1 viral protein R. AB - DNA damage is a universal inducer of cell cycle arrest at the G2 phase. Infection by the human immunodeficiency virus type 1 (HIV-1) also blocks cellular proliferation at the G2 phase. The HIV-1 accessory gene vpr encodes a conserved 96-amino acid protein (Vpr) that is necessary and sufficient for the HIV-1 induced block of cellular proliferation. In the present study, we examined a recently identified DNA damage-signaling protein, the ATM- and Rad3-related protein, ATR, for its potential role in the induction of G2 arrest by Vpr. We show that inhibition of ATR by pharmacological inhibitors, by expression of the dominant-negative form of ATR, or by RNA interference inhibits Vpr-induced cell cycle arrest. As with DNA damage, activation of ATR by Vpr results in phosphorylation of Chk1. This study provides conclusive evidence of activation of the ATR-initiated DNA damage-signaling pathway by a viral gene product. These observations are important toward understanding how HIV infection promotes cell cycle disruption, cell death, and ultimately, CD4+ lymphocyte depletion. PMID- 12738772 TI - Differential terminal fucosylation of N-linked glycans versus protein O fucosylation in leukocyte adhesion deficiency type II (CDG IIc). AB - LAD II/CDG IIc is a rare autosomal recessive disease characterized by a decreased expression of fucosylated antigens on cell surfaces that results in leukocyte adhesion deficiency and severe neurological and developmental abnormalities. Its molecular basis has been identified as a defect in the transporter of GDP-l fucose into the Golgi lumen, which reduces the availability of the substrate for fucosyltransferases. During metabolic radiolabeling experiments using [3H]fucose, LAD II fibroblasts incorporated significantly less radiolabel compared with control cells. However, fractionation and analysis of the different classes of glycans indicated that the decrease in [3H]fucose incorporation is not generalized and is mainly confined to terminal fucosylation of N-linked oligosaccharides. In contrast, the total levels of protein O-fucosylation, including that observed in Notch protein, were unaffected. This finding demonstrates that the decrease in GDP-l-fucose levels in the fibroblast Golgi caused by the LAD II defect does not impair bulk protein O-fucosylation, but severely affects the bulk addition of fucose as a terminal modification of N linked glycans. These data suggest that the severe clinical abnormalities including neurological and developmental ones observed in at least some of the LAD II patients may be related to alteration in recognition systems involving terminal fucose modifications of N-glycans and not be due to a defective O fucosylation of proteins such as Notch. PMID- 12738773 TI - Determinants of non-toxicity in the gastric pathogen Helicobacter pylori. AB - The Helicobacter pylori vacuolating cytotoxin gene, vacA, is naturally polymorphic, the two most diverse regions being the signal region (which can be type s1 or s2) and the mid region (m1 or m2). Previous work has shown which features of vacA make peptic ulcer and gastric cancer-associated type s1/m1 and s1/m2 strains toxic. vacA s2/m2 strains are associated with lower peptic ulcer and gastric cancer risk and are non-toxic. We now define the features of vacA that determine the non-toxicity of these strains. To do this, we deleted parts of vacA and constructed isogenic hybrid strains in which regions of vacA were exchanged between toxigenic and non-toxigenic strains. We showed that a naturally occurring 12-amino acid hydrophilic N-terminal extension found on s2 VacA blocks vacuolating activity as its removal (to make the strain s1-like) confers activity. The mid region of s2/m2 vacA does not cause the non-vacuolating phenotype, but if VacA is unblocked, it confers cell line specificity of vacuolation as in natural s1/m2 strains. Chromosomal replacement of vacA in a non toxigenic strain with vacA from a toxigenic strain confers full vacuolating activity proving that this activity is entirely controlled by elements within vacA. This work defines why H. pylori strains with different vacA allelic structures have differing toxicity and provides a rational basis for vacA typing schemes. PMID- 12738774 TI - Identification of the catalytic residues in family 52 glycoside hydrolase, a beta xylosidase from Geobacillus stearothermophilus T-6. AB - beta-d-Xylosidases (EC 3.2.1.37) are exo-type glycoside hydrolases that hydrolyze short xylooligosaccharides to xylose units. The enzymatic hydrolysis of the glycosidic bond involves two carboxylic acid residues, and their identification, together with the stereochemistry of the reaction, provides crucial information on the catalytic mechanism. Two catalytic mutants of a beta-xylosidase from Geobacillus stearothermophilus T-6 were subjected to detailed kinetic analysis to verify their role in catalysis. The activity of the E335G mutant decreased approximately 106-fold, and this activity was enhanced 103-fold in the presence of external nucleophiles such as formate and azide, resulting in a xylosyl-azide product with an opposite anomeric configuration. These results are consistent with Glu335 as the nucleophile in this retaining enzyme. The D495G mutant was subjected to detailed kinetic analysis using substrates bearing different leaving groups (pKa). The mutant exhibited 103-fold reduction in activity, and the Bronsted plot of log(kcat) versus pKa revealed that deglycosylation is the rate limiting step, indicating that this step was reduced by 103-fold. The rates of the glycosylation step, as reflected by the specificity constant (kcat/Km), were similar to those of the wild type enzyme for hydrolysis of substrates requiring little protonic assistance (low pKa) but decreased 102-fold for those that require strong acid catalysis (high pKa). Furthermore, the pH dependence profile of the mutant enzyme revealed that acid catalysis is absent. Finally, the presence of azide significantly enhanced the mutant activity accompanied with the generation of a xylosyl-azide product with retained anomeric configuration. These results are consistent with Asp495 acting as the acid-base in XynB2. PMID- 12738775 TI - Spinophilin stabilizes cell surface expression of alpha 2B-adrenergic receptors. AB - The third intracellular (3i) loops of the alpha 2A- and alpha 2B-adrenergic receptor (AR) subtypes are critical for retention of these receptors at the basolateral surface of polarized Madin-Darby canine kidney (MDCKII) cells at steady state. The third intracellular loops of the alpha 2A, alpha 2B, and alpha 2C-AR subtypes interact with spinophilin, a multidomain protein that, like the three alpha 2-AR subtypes, is enriched at the basolateral surface of MDCKII cells. The present studies provide evidence that alpha 2-AR interaction with spinophilin contributes to cell surface stabilization of the receptor. We exploited the unique targeting profile of the alpha 2B-AR subtype in MDCKII cells: random delivery to apical and basolateral surfaces with rapid (t(1/2) < or = 60 min) apical versus slower (t(1/2) = 10-12 h) basolateral turnover. Apical delivery of a spinophilin subdomain containing the alpha 2-AR-interacting region (Sp151-483) by fusion with apically targeted p75NTR extended the half-life of alpha 2B-AR at the apical surface to approximately 3.6 h and eliminated the rapid phase (0-60 min) of alpha 2B-AR turnover on that surface. Furthermore, we examined alpha 2B-AR turnover at the surface of mouse embryo fibroblasts derived from wild type (Sp+/+) or spinophilin knock-out (Sp-/-) mice. Two independent experimental approaches demonstrated that agonist-evoked internalization of HA alpha 2B-AR was accelerated in mouse embryo fibroblasts derived from Sp-/- mice. These findings are consistent with the interpretation that endogenous spinophilin contributes to the stabilization of alpha 2B-AR and presumably all three alpha2 AR subtypes at the surface of target cells and may act as a scaffold that could link alpha 2-ARs to proteins interacting with spinophilin via other domains. PMID- 12738776 TI - MAPK-dependent degradation of G protein-coupled receptor kinase 2. AB - G protein-coupled receptor kinase 2 (GRK2) is a key modulator of G protein coupled receptors (GPCR). Altered expression of GRK2 has been described to occur during pathological conditions characterized by impaired GPCR signaling. We have reported recently that GRK2 is rapidly degraded by the proteasome pathway and that beta-arrestin function and Src-mediated phosphorylation are involved in targeting GRK2 for proteolysis. In this report, we show that phosphorylation of GRK2 by MAPK also triggers GRK2 turnover by the proteasome pathway. Modulation of MAPK activation alters the degradation of transfected or endogenous GRK2, and a GRK2 mutant that mimics phosphorylation by MAPK shows an enhanced degradation rate, thus indicating a direct effect of MAPK on GRK2 turnover. Interestingly, MAPK-mediated modulation of wild-type GRK2 stability requires beta-arrestin function and is facilitated by previous phosphorylation of GRK2 on tyrosine residues by c-Src. Consistent with an important physiological role, interfering with this GRK2 degradation process results in altered GPCR responsiveness. Our data suggest that both c-Src and MAPK-mediated phosphorylation would contribute to modulate GRK2 degradation, and put forward the existence of new feedback mechanisms connecting MAPK cascades and GPCR signaling. PMID- 12738777 TI - TDAG51 is induced by homocysteine, promotes detachment-mediated programmed cell death, and contributes to the cevelopment of atherosclerosis in hyperhomocysteinemia. AB - Hyperhomocysteinemia is an independent risk factor for cardiovascular disease and accelerates atherosclerosis in apoE-/- mice. Despite the observations that homocysteine causes endoplasmic reticulum (ER) stress and programmed cell death (PCD) in cultured human vascular endothelial cells, the cellular factors responsible for this effect and their relevance to atherogenesis have not been completely elucidated. We report here that homocysteine induces the expression of T-cell death-associated gene 51 (TDAG51), a member of the pleckstrin homology related domain family, in cultured human vascular endothelial cells. This effect was observed for other ER stress-inducing agents, including dithiothreitol and tunicamycin. TDAG51 expression was attenuated in homozygous A/A mutant eukaryotic translation initiation factor 2 alpha mouse embryonic fibroblasts treated with homocysteine or tunicamycin, suggesting that ER stress-induced phosphorylation of eukaryotic translation initiation factor 2 alpha is required for TDAG51 transcriptional activation. Transient overexpression of TDAG51 elicited significant changes in cell morphology, decreased cell adhesion, and promoted detachment-mediated PCD. In support of these in vitro findings, TDAG51 expression was increased and correlated with PCD in the atherosclerotic lesions from apoE-/- mice fed hyperhomocysteinemic diets, compared with mice fed a control diet. Collectively, these findings provide evidence that TDAG51 is induced by homocysteine, promotes detachment-mediated PCD, and contributes to the development of atherosclerosis observed in hyperhomocysteinemia. PMID- 12738779 TI - Identification of peptide substrates for human MMP-11 (stromelysin-3) using phage display. AB - The MMP-11 proteinase, also known as stromelysin-3, probably plays an important role in human cancer because MMP-11 is frequently overexpressed in human tumors and MMP-11 levels affect tumorogenesis in mice. Unlike other MMPs, however, human MMP-11 does not cleave extracellular matrix proteins, such as collagen, laminin, fibronectin, and elastin. To help identify physiologic MMP-11 substrates, a phage display library was used to find peptide substrates for MMP-11. One class of peptides containing 26 members had the consensus sequence A(A/Q)(N/A) downward arrow (L/Y)(T/V/M/R)(R/K), where downward arrow denotes the cleavage site. This consensus sequence was similar to that for other MMPs, which also cleave peptides containing Ala in position 3, Ala in position 1, and Leu/Tyr in position 1', but differed from most other MMP substrates in that proline was rarely found in position 3 and Asn was frequently found in position 1. A second class of peptides containing four members had the consensus sequence G(G/A)E downward arrow LR. Although other MMPs also cleave peptides with these residues, other MMPs prefer proline at position 3 in this sequence. In vitro assays with MMP-11 and representative peptides from both classes yielded modest kcat/Km values relative to values found for other MMPs with their preferred peptide substrates. These reactions also showed that peptides with proline in position 3 were poor substrates for MMP-11. A structural basis for the lower kcat/Km values of human MMP-11, relative to other MMPs, and poor cleavage of position 3 proline substrates by MMP-11 is provided. Taken together, these findings explain why MMP 11 does not cleave most other MMP substrates and predict that MMP-11 has unique substrates that may contribute to human cancer. PMID- 12738778 TI - The activation of matriptase requires its noncatalytic domains, serine protease domain, and its cognate inhibitor. AB - The activation of matriptase requires proteolytic cleavage at a canonical activation motif that converts the enzyme from a one-chain zymogen to an active, two-chain protease. In this study, matriptase bearing a mutation in its catalytic triad was unable to undergo this activational cleavage, suggesting that the activating cleavage occurs via a transactivation mechanism where interaction between matriptase zymogen molecules leads to activation of the protease. Using additional point and deletion mutants, we showed that activation of matriptase requires proteolytic processing at Gly-149 in the SEA domain of the protease, glycosylation of the first CUB domain and the serine protease domain, and intact low density lipoprotein receptor class A domains. Its cognate inhibitor, hepatocyte growth factor activator inhibitor-1, may also participate in the activation of matriptase, based on the observation that matriptase activation did not occur when the protease was co-expressed with hepatocyte growth factor activator inhibitor-1 mutated in its low density lipoprotein receptor class A domain. These results suggest that besides matriptase catalytic activity, matriptase activation requires post-translational modification of the protease, intact noncatalytic domains, and its cognate inhibitor. PMID- 12738780 TI - Gain of glutaminase function in mutants of the ammonia-specific frog carbamoyl phosphate synthetase. AB - Depending on their physiological role, carbamoyl phosphate synthetases (CPSs) use either glutamine or free ammonia as the nitrogen donor for carbamoyl phosphate synthesis. Sequence analysis of known CPSs indicates that, regardless of whether they are ammonia- or glutamine-specific, all CPSs contain the structural equivalent of a triad-type glutamine amidotransferase (GAT) domain. In ammonia specific CPSs, such as those of rat or human, the catalytic inactivity of the GAT domain can be rationalized by the substitution of the Triad cysteine residue by serine (1). The ammonia-specific CPS of Rana catesbeiana (fCPS) presents an interesting anomaly in that, despite its retention of the entire catalytic triad (2) and almost all other residues conserved in Triad GATs, it is unable to utilize glutamine as a nitrogen-donating substrate (3). Based on our earlier work with the glutamine-utilizing E. coli CPS (eCPS), we have targeted residues Lys258 and Glu261 in the fCPS GAT domain as critical for preventing GAT function. Previously we have shown that substitution of the corresponding residues in eCPS by their fCPS counterparts (Leu --> Lys and Gln --> Glu) resulted in complete loss of GAT function in eCPS (3). To examine the role of these residues in the fCPS GAT component, we have cloned the full-length fCPS gene from R. catesbeiana liver. Here we report the first heterologous expression of an ammonia-specific CPS and show that a single mutation of the frog enzyme, K258L, yields a gain of glutaminase function. PMID- 12738781 TI - Identification of a consensus motif for Plk (Polo-like kinase) phosphorylation reveals Myt1 as a Plk1 substrate. AB - Plk1 (Polo-like kinase 1), an evolutionarily conserved serine/threonine kinase, is crucially involved in multiple events during the M phase. Here we have identified a consensus phosphorylation sequence for Plk1, by testing the ability of systematically mutated peptides derived from human Cdc25C to serve as a substrate for Plk1. The obtained results show that a hydrophobic amino acid at position +1 carboxyl-terminal of phosphorylated Ser/Thr and an acidic amino acid at position -2 are important for optimal phosphorylation by Plk1. We have then found that Myt1, an inhibitory kinase for MPF, has a number of putative phosphorylation sites for Plk1 in its COOH-terminal portion. While wild-type Myt1 (Myt1-WT) served as a good substrate for Plk1 in vitro, a mutant Myt1 (Myt1-4A), in which the four putative phosphorylation sites are replaced by alanines, did not. In nocodazole-treated cells, Myt1-WT, but not Myt1-4A, displayed its mobility shift in gel electrophoresis, due to phosphorylation. These results suggest that Plk1 phosphorylates Myt1 during M phase. Thus, this study identifies a novel substrate for Plk1 by determining a consensus phosphorylation sequence by Plk1. PMID- 12738782 TI - Intracellular coupling via limiting calmodulin. AB - Measurements of cellular Ca2+-calmodulin concentrations have suggested that competition for limiting calmodulin may couple calmodulin-dependent activities. Here we have directly tested this hypothesis. We have found that in endothelial cells the amount of calmodulin bound to nitric-oxide synthase and the catalytic activity of the enzyme both are increased approximately 3-fold upon changes in the phosphorylation status of the enzyme. Quantitative immunoblotting indicates that the synthase can bind up to 25% of the total cellular calmodulin. Consistent with this, simultaneous determinations of the free Ca2+ and Ca2+-calmodulin concentrations in these cells performed using indo-1 and a fluorescent calmodulin biosensor (Kd = 2 nm) indicate that increased binding of calmodulin to the synthase is associated with substantial reductions in the Ca2+-calmodulin concentrations produced and an increase in the [Ca2+]50 for formation of the calmodulin-biosensor complex. The physiological significance of these effects is confirmed by a corresponding 40% reduction in calmodulin-dependent plasma membrane Ca2+ pump activity. An identical reduction in pump activity is produced by expression of a high affinity (Kd = 0.3 nm) calmodulin biosensor, and treatment to increase calmodulin binding to the synthase then has no further effect. This suggests that the observed reduction in pump activity is due specifically to reduced calmodulin availability. Increases in synthase activity thus appear to be coupled to decreases in the activities of other calmodulin targets through reductions in the size of a limiting pool of available calmodulin. This exemplifies what is likely to be a ubiquitous mechanism for coupling among diverse calmodulin-dependent activities. PMID- 12738783 TI - Regulation of multidrug resistance in cancer cells by hyaluronan. AB - Multidrug resistance in cancer cells is often due to ATP-dependent efflux pumps, but is also linked to alterations in cell survival and apoptotic signaling pathways. We have found previously that perturbation of hyaluronan-tumor cell interaction by treatment with hyaluronan oligosaccharides suppresses the phosphoinositide 3-kinase/Akt cell survival signaling pathway in cancer cells and reduces tumor growth in vivo. Here we find that these oligomers suppress both the MAP kinase and phosphoinositide 3-kinase pathways in multidrug resistant tumor cells and sensitize these cells to a variety of chemotherapeutic drugs. On the other hand, increased hyaluronan production induces resistance in drug-sensitive tumor cells. Likewise, increased expression of emmprin, which is a glycoprotein that is present on the surface of most malignant cancer cells and that stimulates hyaluronan production, also induces increased resistance. Thus, perturbation of hyaluronan signaling may provide a dual therapeutic role, since it has intrinsic suppressive effects on tumor growth as well as sensitizing cancer cells to chemotherapeutic agents. PMID- 12738784 TI - Transcriptional activation of the human brain-derived neurotrophic factor gene promoter III by dopamine signaling in NT2/N neurons. AB - We have identified a functional cAMP-response element (CRE) in the human brain derived neurotrophic factor (BDNF) gene promoter III and established that it participated in the modulation of BDNF expression in NT2/N neurons via downstream signaling from the D1 class of dopamine (DA) receptors. The up-regulation of BDNF expression, in turn, produced neuroprotective signals through receptor tyrosine kinase B (TrkB) and promoted cell survival under the conditions of oxygen and glucose deprivation. To our knowledge this is the first evidence showing the presence of a functional CRE in the human BDNF gene and the role of DA signaling in establishing transcriptional competence of CRE in post-mitotic NT2/N neurons. This ability of DA to regulate the expression of the BDNF survival factor has a profound significance for the nigrostriatal pathway, because it indicates the existence of a feedback loop between the neutrophin, which promotes both the maturation and survival of dopaminergic neurons, and the neurotransmitter, which the mature neurons ultimately produce and release. PMID- 12738786 TI - Serine-arginine-rich protein p30 directs alternative splicing of glucocorticoid receptor pre-mRNA to glucocorticoid receptor beta in neutrophils. AB - Glucocorticoid (GC) insensitivity is a major clinical challenge in the treatment of many inflammatory diseases. It has been shown previously that GC insensitivity, in several inflammatory cell types, is due to an overabundance of the beta isoform of the glucocorticoid receptor (GCRbeta) relative to the ligand binding isoform, GCRalpha. GCRbeta functions as a dominant inhibitor of GCRalpha action. A number of GCR isoforms are created from the same pre-mRNA transcript via alternative splicing, and the factor or factors that control alternative splicing of GCR pre-mRNA are of great importance. In the current study, we have identified the predominant alternative splicing factor present in human neutrophils, which is known to be exceptionally GC-insensitive. The predominant alternative splicing factor in neutrophils is SRp30c, which is one of several highly conserved serine-arginine-rich (SR) proteins that are involved in both constitutive and alternative splicing in eukaryotic cells. Inhibition of SRp30c expression with antisense oligonucleotide strongly inhibited expression of GCRbeta and stimulated expression of GCRalpha. Antisense molecules targeted to other SR proteins had no effect. Our data indicate that SRp30c is necessary for alternative splicing of the GCR pre-mRNA to create mRNA encoding GCRbeta. PMID- 12738785 TI - Amino acids Glu323, Tyr324, Glu330, and Val331 of factor Va heavy chain are essential for expression of cofactor activity. AB - We have recently demonstrated that amino acid region 323-331 of factor Va heavy chain (9 amino acids, AP4') contains a binding site for factor Xa (Kalafatis, M., and Beck, D. O. (2002) Biochemistry 41, 12715-12728). To ascertain which amino acids within this region are important for the effector and receptor properties of the cofactor with respect to factor Xa, we have synthesized three overlapping peptides (5 amino acids each) spanning the amino acid region 323-331 and tested them for their effect on prothrombinase complex assembly and function. Peptide containing amino acids 323EYFIA327 alone was found to increase the catalytic efficiency of factor Xa but had no effect on the fluorescent anisotropy of active site-labeled factor Xa (human factor Xa labeled in the active site with Oregon Green 488; [OG488]-EGR-hXa). In contrast, peptide containing the sequence 327AAEEV331 was found to interact with [OG488]-EGR-hXa with half-maximal saturation reached at approximately 150 microm, but it was unable to produce a cofactor effect on factor Xa. Peptide 325FIAAE329 inhibited prothrombinase activity and was able to partially decrease the fluorescent anisotropy of [OG488] EGR-hXa but could not increase the catalytic efficiency of factor Xa with respect to prothrombin. A control peptide with the sequence FFFIA did not increase the catalytic efficiency of factor Xa, whereas a peptide with the sequence AAEMI was impaired in its capability to interact with [OG488]-EGR-hXa. Two mutant recombinant factor Va molecules (Glu323 --> Phe/Tyr324 --> Phe, factor VaFF; Glu330 --> Met/Val331 --> Ile, factor VaMI) showed impaired cofactor activity when used at limiting cofactor concentration, whereas the quadruple mutant (Glu323 --> Phe/Tyr324 --> Phe and Glu330 --> Met/Val331 --> Ile, factor VaFF/MI) had no cofactor activity under similar experimental conditions. Our data demonstrate that amino acid residues Glu323, Tyr324, Glu330, and Val331 of factor Va heavy chain are critical for expression of factor Va cofactor activity. PMID- 12738787 TI - Nuclear factor of activated T cells 2 transactivation in mast cells: a novel isoform-specific transactivation domain confers unique FcepsilonRI responsiveness. AB - Murine nuclear factor of activated T cells (NFAT)2.alpha/beta differ by 42 and 28 unique amino-terminal amino acids and are differentially expressed. Both isoforms share conserved domains that regulate DNA-binding and subcellular localization. A genetic "one-hybrid" assay was used to define two distinct transactivation (TA) domains: in addition to a conserved TAD present in both isoforms, a second, novel TAD exists within the beta-specific amino terminus. Pharmacologic inhibitors Go6976 and rottlerin demonstrate that both conventional and novel protein kinase C (PKC) family members regulate endogenous mast cell NFAT activity, and NFAT2 TA. Overexpression of dominant active PKC (which has been implicated in immune receptor signaling) induces NFAT2.alpha/beta TA. Mutations within the smallest PKC-responsive transactivation domain demonstrate that the PKC effect is at least partially indirect. Significantly, the beta-specific domain confers greater ability to TA in response to treatment with phorbol 12-myristate 13 acetate/ionomycin or lipopolysaccharide, and unique sensitivity to FcepsilonRI signaling. Accordingly, overexpression of NFAT2.beta results in significantly greater NFAT- and interleukin-4 reporter activity than NFAT2.alpha. These results suggest that whereas NFAT2 isoforms may share redundant DNA-binding preferences, there are specialized functional consequences of their isoform-specific domains. PMID- 12738788 TI - Full activation of estrogen receptor alpha activation function-1 induces proliferation of breast cancer cells. AB - The effects of estrogen and anti-estrogen are mediated through the estrogen receptors (ER) alpha and beta, which function as ligand-induced transcriptional factors. Recently, one of the phthalate esters, n-butylbenzyl phthalate (BBP), has been shown to induce estrogen receptor-mediated responses. By using the truncated types of ER mutants, we revealed that activation function-1 (AF-1) activity was necessary for the BBP-dependent transactivation function of ERalpha. AF-1 is also known to be responsible for the partial agonistic activity of tamoxifen. Whereas tamoxifen exhibits an anti-estrogenic effect on proliferation of the MCF-7 breast cancer cell line, BBP showed an estrogenic effect on MCF-7 to stimulate proliferation. In vivo and in vitro binding assays revealed that whereas 4-hydroxytamoxifen (OHT) induced binding of ERalpha to both an AF-1 coactivator complex (p68/p72 and p300) and corepressor complexes (N-CoR/SMRT), BBP selectively enhanced the binding to the AF-1 coactivators. We also showed that the transcriptional activity of OHT-bound ERalpha was modulated by the ratio between the AF-1 coactivator and corepressor complexes. Expression of a dominant negative type of N-CoR inhibited the interaction between OHT-bound ERalpha and N CoR/SMRT and enhanced the transcriptional activity of OHT-bound ERalpha. Furthermore, the cell growth of MCF-7 stably expressing the dominant-negative type of N-CoR was enhanced by the addition of OHT. These results indicated that fully activated AF-1 induces the stimulation of breast cancer growth and that the ratio between AF-1 coactivators and corepressors plays a key role to prevent proliferation of tumor by tamoxifen. PMID- 12738789 TI - Bcl-xL mediates a survival mechanism independent of the phosphoinositide 3 kinase/Akt pathway in prostate cancer cells. AB - Among various molecular strategies by which prostate cancer cells evade apoptosis, phosphoinositide 3-kinase (PI3K)/Akt signaling represents a dominant survival pathway. However, different prostate cancer cell lines such as LNCaP and PC-3 display differential sensitivity to the apoptotic effect of PI3K inhibition in serum-free media, reflecting the heterogeneous nature of prostate cancer in apoptosis regulation. Whereas both cell lines are equally susceptible to LY294002 mediated Akt dephosphorylation, only LNCaP cells default to apoptosis, as evidenced by DNA fragmentation and cytochrome c release. In PC-3 cells, Akt deactivation does not lead to cytochrome c release, suggesting that the intermediary signaling pathway is short-circuited by an antiapoptotic factor. This study presents evidence that Bcl-xL overexpression provides a distinct survival mechanism that protects PC-3 cells from apoptotic signals emanating from PI3K inhibition. First, the Bcl-xL/BAD ratio in PC-3 cells is at least an order of magnitude greater than that of LNCaP cells. Second, ectopic expression of Bcl xL protects LNCaP cells against LY294002-induced apoptosis. Third, antisense down regulation of Bcl-xL sensitizes PC-3 cells to the apoptotic effect of LY294002. The physiological relevance of this Bcl-xL-mediated survival mechanism is further underscored by the protective effect of serum on LY294002-induced cell death in LNCaP cells, which is correlated with a multifold increase in Bcl-xL expression. In contrast to Bcl-xL, Bcl-2 expression levels are similar in both cells lines, and do not respond to serum stimulation, suggesting that Bcl-2 may not play a physiological role in antagonizing apoptosis signals pertinent to BAD activation in prostate cancer cells. PMID- 12738790 TI - Protection of renal epithelial cells against oxidative injury by endoplasmic reticulum stress preconditioning is mediated by ERK1/2 activation. AB - We investigated the role of the endoplasmic reticulum (ER) stress response in intracellular Ca2+ regulation, MAPK activation, and cytoprotection in LLC-PK1 renal epithelial cells in an attempt to identify the mechanisms of protection afforded by ER stress. Cells preconditioned with trans-4,5-dihydroxy-1,2 dithiane, tunicamycin, thapsigargin, or A23187 expressed ER stress proteins and were resistant to subsequent H2O2-induced cell injury. In addition, ER stress preconditioning prevented the increase in intracellular Ca2+ concentration that normally follows H2O2 exposure. Stable transfection of cells with antisense RNA targeted against GRP78 (pkASgrp78 cells) prevented GRP78 induction, disabled the ER stress response, sensitized cells to H2O2-induced injury, and prevented the development of tolerance to H2O2 that normally occurs with preconditioning. ERK and JNK were transiently (30-60 min) phosphorylated in response to H2O2. ER stress-preconditioned cells had more ERK and less JNK phosphorylation than control cells in response to H2O2 exposure. Preincubation with a specific inhibitor of JNK activation or adenoviral infection with a construct that encodes constitutively active MEK1, the upstream activator of ERKs, also protected cells against H2O2 toxicity. In contrast, the pkASgrp78 cells had less ERK and more JNK phosphorylation upon H2O2 exposure. Expression of constitutively active ERK also conferred protection on native as well as pkAS-grp78 cells. These results indicate that GRP78 plays an important role in the ER stress response and cytoprotection. ER stress preconditioning attenuates H2O2-induced cell injury in LLC-PK1 cells by preventing an increase in intracellular Ca2+ concentration, potentiating ERK activation, and decreasing JNK activation. Thus, the ER stress response modulates the balance between ERK and JNK signaling pathways to prevent cell death after oxidative injury. Furthermore, ERK activation is an important downstream effector mechanism for cellular protection by ER stress. PMID- 12738791 TI - Temperature-modulated diversity of TRPV4 channel gating: activation by physical stresses and phorbol ester derivatives through protein kinase C-dependent and independent pathways. AB - The TRPV4 calcium-permeable channel was cloned from mouse kidney M-1 cells, and the effect of temperature modulation on channel gating/activation by physical and chemical signals was evaluated. A TRPV4 cDNA construct with a C-terminal V5 epitope was stably transfected into human embryonic kidney (HEK) 293 and Chinese hamster ovary cells resulting in high levels of expression at the plasma membrane. Channel activation was assessed from changes in calcium influx (fura-2 fluorescence measurements) or whole cell currents (patch clamp analysis). At room temperature (22-24 degrees C), exposure of TRPV4-transfected cells to hypotonic medium (225 mOsm/liter) or a non-protein kinase C (PKC)-activating phorbol ester derivative, 4alpha-phorbol 12,13-decanoate (100 nm), induces modest channel activation, whereas phorbol 12-myristate 13-acetate (100 nm), a PKC-activating phorbol ester, and shear stress (3-20 dyne/cm2) had minimal or no effect on channel activation. In contrast, at elevated temperatures (37 degrees C) the channel was rapidly activated by all stimuli. Inhibition of PKC by calphostin C (50 nm) or staurosporine (500 nm) abolished phorbol 12-myristate 13-acetate induced activation of the channel without affecting the response to other stimuli. Ruthenium red (1 microm) effectively blocked the channel activity by all stimuli. It is concluded that temperature is a critical modulator of TRPV4 channel gating, leading to activation of the channel by a diverse range of microenvironmental chemical and physical signals utilizing a least two transduction pathways, one PKC-dependent and one PKC-independent. The convergence of multiple signals and transduction pathways on the same channel indicate that the channel functions as a molecular integrator of microenvironmental chemical and physical signals. PMID- 12738793 TI - Requirement of the p130CAS-Crk coupling for metastasis suppressor KAI1/CD82 mediated inhibition of cell migration. AB - KAI1/CD82 protein is a member of the tetraspanin superfamily and has been rediscovered as a cancer metastasis suppressor. The mechanism of KAI1/CD82 mediated suppression of cancer metastasis remains to be established. In this study, we found that migration of the metastatic prostate cancer cell line Du145 was substantially inhibited when KAI1/CD82 was expressed. The expression of focal adhesion kinase (FAK) and Lyn, a Src family tyrosine kinase and substrate of FAK, was up-regulated at both RNA and protein levels upon KAI1/CD82 expression. The activation of FAK and Lyn, however, remained unchanged in Du145-KAI1/CD82 cells. As a downstream target of FAK-Lyn signaling, the p130CAS (Crk-associated substrate) protein was decreased upon the expression of KAI1/CD82. Consequently, less p130CAS-CrkII complex, which functions as a "molecular switch" in cell motility, was formed in Du145-KAI1/CD82 cells. To confirm that the p130CAS-CrkII complex is indeed important for the motility inhibition by KAI1/CD82, overexpression of p130CAS in Du145-KAI1/CD82 cells increased the formation of p130CAS-CrkII complex and largely reversed the KAI1/CD82-mediated inhibition of cell motility. Taken together, our studies indicate the following: 1) signaling of FAK-Lyn-p130CAS-CrkII pathway is altered in KAI1/CD82-expressing cells, and 2) p130CAS-CrkII coupling is required for KAI1/CD82-mediated suppression of cell motility. PMID- 12738792 TI - RNA-structural mimicry in Escherichia coli ribosomal protein L4-dependent regulation of the S10 operon. AB - Ribosomal protein L4 regulates the 11-gene S10 operon in Escherichia coli by acting, in concert with transcription factor NusA, to cause premature transcription termination at a Rho-independent termination site in the leader sequence. This process presumably involves L4 interaction with the leader mRNA. Here, we report direct, specific, and independent binding of ribosomal protein L4 to the S10 mRNA leader in vitro. Most of the binding energy is contributed by a small hairpin structure within the leader region, but a 64-nucleotide sequence is required for the bona fide interaction. Binding to the S10 leader mRNA is competed by the 23 S rRNA L4 binding site. Although the secondary structures of the mRNA and rRNA binding sites appear different, phosphorothioate footprinting of the L4-RNA complexes reveals close structural similarity in three dimensions. Mutational analysis of the mRNA binding site is compatible with the structural model. In vitro binding of L4 induces structural changes of the S10 leader RNA, providing a first clue for how protein L4 may provoke transcription termination. PMID- 12738794 TI - Chronic stimulation of D1 dopamine receptors in human SK-N-MC neuroblastoma cells induces nitric-oxide synthase activation and cytotoxicity. AB - Elevated synaptic levels of dopamine may induce striatal neurodegeneration in l DOPA-unresponsive parkinsonism subtype of multiple system atrophy (MSA-P subtype), multiple system atrophy, and methamphetamine addiction. We examined the participation of dopamine and D1 dopamine receptors in the genesis of postsynaptic neurodegeneration. Chronic treatment of human SK-N-MC neuroblastoma cells with dopamine or H2O2 increased NO production and accelerated cytotoxicity, as indexed by enhanced nitrite levels and cell death. The antioxidant sodium metabisulfite or SCH 23390, a D1 dopamine receptor-selective antagonist, partially blocked dopamine effects but together ablated dopamine-mediated cytotoxicity, indicating the participation of both autoxidation and D1 receptor stimulation. Direct activation of D1 dopamine receptors with SKF R-38393 caused cytotoxicity, which was refractory to sodium metabisulfite. Dopamine and SKF R 38393 induced overexpression of the nitric-oxide synthase (NOS) isoforms neuronal NOS, inducible NOS (iNOS), and endothelial NOS in a protein kinase A-dependent manner. Functional studies showed that approximately 60% of total NOS activity was due to activation of iNOS. The NOS inhibitor N(G)-nitro-l-arginine methyl ester and genistein, wortmannin, or NF-kappaB SN50, inhibitors of protein tyrosine kinases phosphatidylinositol 3-kinase and NF-kappaB, respectively, reduced nitrite production by dopamine and SKF R-38393 but were less effective in attenuating H2O2-mediated effects. In rat striatal neurons, dopamine and SKF R 38393, but not H2O2, accelerated cell death through increased expression of neuronal NOS and iNOS but not endothelial NOS. These data demonstrate a novel pathway of dopamine-mediated postsynaptic oxidative stress and cell death through direct activation of NOS enzymes by D1 dopamine receptors and its associated signaling pathways. PMID- 12738795 TI - Membrane translocation of protein kinase Ctheta during T lymphocyte activation requires phospholipase C-gamma-generated diacylglycerol. AB - Protein kinase C (PKC) is the only PKC isoform recruited to the immunological synapse after T cell receptor stimulation, suggesting that its activation mechanism differs from that of the other isoforms. Previous studies have suggested that this selective PKC recruitment may operate via a Vav-regulated, cytoskeletal-dependent mechanism, independent of the classical phospholipase C/diacylglycerol pathway. Here, we demonstrate that, together with tyrosine phosphorylation of PKC in the regulatory domain, binding of phospholipase C dependent diacylglycerol is required for PKC recruitment to the T cell synapse. In addition, we demonstrate that diacylglycerol kinase alpha-dependent diacylglycerol phosphorylation provides the negative signal required for PKC inactivation, ensuring fine control of the T cell activation response. PMID- 12738796 TI - Cross-talk between JNK/SAPK and ERK/MAPK pathways: sustained activation of JNK blocks ERK activation by mitogenic factors. AB - Mixed lineage kinases (MLKs) are a family of serine/threonine kinases that function in the SAPK signaling cascade. MLKs activate JNK/SAPK in vivo by directly phosphorylating and activating the JNK kinase SEK-1 (MKK4 and -7). Importantly, the MLK member MLK3/SPRK has been shown recently to be a direct target of ceramide and tumor necrosis factor-alpha (TNF-alpha) and to mediate the TNF-alpha and ceramide-induced JNK activation in Jurkat cells. Here we report that MLK3 can phosphorylate and activate MEK-1 directly in vitro and also can induce MEK phosphorylation on its activation sites in vivo in COS-7 cells. Surprisingly, this induction of MEK phosphorylation does not result in ERK activation in vivo. Rather, in cells expressing active MLK3, ERK becomes resistant to activation by growth factors and mitogens. This restriction in ERK activation requires MLK3 kinase activity, is independent of Raf activation, and is reversed by JNK pathway inhibition either at the level of SEK-1, JNK, or Jun. These results demonstrate that sustained JNK activation uncouples ERK activation from MEK in a manner requiring Jun-mediated gene transcription. This in turn points to the existence of a negative cross-talk relationship between the stress activated JNK pathway and the mitogen-activated ERK pathway. Thus, our findings imply that some of the biological functions of JNK activators, such as TNF-alpha and ceramide, may be attributed to their ability to block cell responses to growth and survival factors acting through the ERK/MAPK pathway. PMID- 12738797 TI - Serotonin is a novel survival factor of cardiomyocytes: mitochondria as a target of 5-HT2B receptor signaling. AB - Identification of factors regulating cardiomyocyte survival and growth is important to understand the pathogenesis of congenital heart diseases. Little is known about the molecular mechanism of cardiac functions triggered by serotonin. The link between signaling circuitry of external stimuli and the mitochondrial apoptotic machinery is of wide interest in cardiac diseases. Using cultured cardiomyocytes and 5-hydroxytryptamine (5-HT)2B-receptor knockout mice as an animal model of dilated cardiomyopathy, for the first time we show that serotonin via the Gq-coupled 5-HT2B-receptor protect cardiomyocytes against serum deprivation-induced apoptosis as manifested by DNA fragmentation, nuclear chromatin condensation, and TUNEL labeling. Serotonin prevents cytochrome c release and caspase-9 and -3 activation after serum deprivation via cross-talks between phosphatidylinositol-3 kinase/Akt and extracellular signal-regulated kinase (ERK) 1/2 signaling pathways. Serotonin binding to 5-HT2B-receptor activates ERK kinases to inhibit Bax expression induced by serum deprivation. Serotonin via phosphatidylinositol-3 kinase/Akt can activate NF-kappaB that is required for the regulation of the mitochondrial adenine nucleotide translocator (ANT-1). Parallel to these observations, ultrastructural analysis in the 5-HT2B receptor knockout mice heart revealed pronounced mitochondrial defects in addition to altered mitochondrial enzyme activities (cytochrome oxidase and succinate dehydrogenase) and ANT-1 and Bax expressions. These findings identify 5 HT as a novel survival factor targeting mitochondria in cardiomyocytes. PMID- 12738799 TI - Cyclooxygenase-2-deficient mice are resistant to endotoxin-induced inflammation and death. AB - Sepsis is a systemic inflammatory response to a blood-borne infection that is associated with an extremely high rate of morbidity and mortality. The present study investigates the role of cyclooxygenase (COX)-2 in host responses to bacterial endotoxemia. After administration of Escherichia coli lipopolysaccharide, 50% of wild-type mice die within 96 h. COX-2 deficient mice displayed a dramatic improvement in survival with reduced leukocyte infiltration into critical organs (kidneys and lungs) and a blunted and delayed induction of the cytokine inducible genes nitric oxide synthase 2 and heme oxygenase-1. Translocation and activation of transcription factors important for signaling events during an inflammatory response, such as nuclear factor (NF)-kappaB, were also markedly reduced. While the absence of COX-2 did not alter the induction of several pro-inflammatory cytokines in tissue macrophages, induction of the anti inflammatory cytokine IL-10 was exaggerated. Administration of IL-10 to wild-type mice reduced NF-kappaB activation. Taken together, our data suggest that COX-2 deficient mice are resistant to many of the detrimental consequences of endotoxemia. These beneficial effects occur, in part, by a compensatory increase in IL-10 that counterbalances the pro-inflammatory host response to endotoxemia. PMID- 12738801 TI - Oxygen-regulated expression of the Wilms' tumor suppressor Wt1 involves hypoxia inducible factor-1 (HIF-1). AB - The Wilms' tumor gene Wt1 is unique among tumor suppressors because of its requirement for the development of certain organs. We recently described de novo expression of Wt1 in myocardial blood vessels of ischemic rat hearts. The purpose of this study was to analyze the mechanism(s) of hypoxic/ischemic induction of Wt1. We show here that Wt1 mRNA and protein is up-regulated in the heart and kidneys of rats exposed to normobaric hypoxia (8% O2). Ectopic Wt1 immunoreactivity was detected in renal tubules of hypoxic rats, which also expressed the antiapoptotic protein Bcl-2 and contained significantly fewer TUNEL positive cells than in normoxic kidneys. Wt1 expression was enhanced in the osteosarcoma line U-2OS and in Reh lymphoblast cells that were grown either at 1% O2 or in the presence of CoCl2 and desferrioxamine, respectively. The promoter of the Wt1 gene was capable of mediating expression of a luciferase reporter in response to hypoxia. We identified a hypoxia-responsive element in the Wt1 sequence that bound to hypoxia-inducible factor-1 (HIF-1) and was required for activation of the Wt1 promoter by CoCl2 and HIF-1. These findings demonstrate that Wt1 expression can be stimulated by hypoxia, which involves activation of the Wt1 promoter by HIF-1. PMID- 12738800 TI - Caspase-8 expression and proteolysis in human brain after severe head injury. AB - Programmed cell death involves a complex and interrelated cascade of cysteine proteases termed caspases that are synthesized as inactive zymogens, which are proteolytically processed to active enzymes. Caspase-8 is an initiator caspase that becomes activated when Fas death receptor-Fas ligand (FasL) coupling on the cell surface leads to coalescence of a "death complex" perpetuating the programmed cell death cascade. In this study, brain tissue samples removed from adult patients during the surgical management of severe intracranial hypertension after traumatic brain injury (TBI; n=17) were compared with postmortem control brain tissue samples (n=6). Caspase-8 mRNA was measured by semiquantitative reverse transcription and polymerase chain reaction, and caspase-8 protein was examined by Western blot and immunocytochemistry. Fas and FasL were also examined using Western blot. Caspase-8 mRNA and protein were increased in TBI patients vs. controls, and caspase-8 protein was predominately expressed in neurons. Proteolysis of caspase-8 to 20-kDa fragments was seen only in TBI patients. Fas was also increased after TBI vs. control and was associated with relative levels of caspase-8, supporting formation of a death complex. These data identify additional steps in the programmed cell death cascade involving Fas death receptors and caspase-8 after TBI in humans. PMID- 12738802 TI - Beating is necessary for transdifferentiation of skeletal muscle-derived cells into cardiomyocytes. AB - Cell transplantation could be a potential therapy for heart damage. Skeletal myoblasts have been expected to be a good cell source for autologous transplantation; however, the safety and efficacy of their transplantation are still controversial. Recent studies have revealed that skeletal muscle possesses the stem cell population that is distinct from myoblasts. To elucidate whether skeletal muscle stem cells can transdifferentiate into cardiomyocytes, we cocultured skeletal muscle cells isolated from transgenic mice expressing green fluorescent protein with cardiomyocytes of neonatal rats. Skeletal muscle-derived cells expressed cardiac-specific proteins such as cardiac troponin T and atrial natriuretic peptide as well as cardiac-enriched transcription factors such as Nkx2E (formerly called Csx/Nkx2.5) and GATA4 by coculture with cardiomyocytes. Skeletal muscle-derived cells also expressed cadherin and connexin 43 at the junctions with neighboring cardiomyocytes. Cardiomyocyte-like action potentials were recorded from beating skeletal muscle-derived cells. Treatment of nifedipine or culture in Ca2+-free media suppressed contraction of cardiomyocytes and inhibited skeletal muscle cells to express cardiac-specific proteins. Cyclic stretch completely restored this inhibitory effect. These results suggest that some part of skeletal muscle cells can transdifferentiate into cardiomyocytes and that direct cell-to-cell contact and contraction of neighboring cardiomyocytes are important for the transdifferentiation. PMID- 12738803 TI - Transduction of biologically active motifs of the small heat shock-related protein HSP20 leads to relaxation of vascular smooth muscle. AB - Activation of cyclic nucleotide-dependent signaling pathways leads to phosphorylation of the small heat shock-related protein, HSP20, on serine 16, and relaxation of vascular smooth muscle. In this study, we used an enhanced protein transduction domain (PTD) sequence to deliver HSP20 phosphopeptide analogs into porcine coronary artery. The transduction of phosphoHSP20 analogs led to dose dependent relaxation of coronary artery smooth muscle. Peptides containing the protein transduction domain coupled to a random orientation of the same amino acids did not. Direct fluorescence microscopy of arterial rings incubated with fluorescein isothiocyanate (FITC)-PTD or FITC-PTD-HSP20 peptides showed a diffuse peptide uptake. Mass spectrometric immunoassays (MSIAs) of smooth muscle homogenates were used to determine whether the phosphopeptide analogs affected the phosphorylation of endogenous HSP20. Treatment with the phosphodiesterase inhibitor papaverine led to a mass shift of 80 Da. However, there was no mass shift of HSP20 in muscles treated with phosphoHSP20 analogs. This suggests that the PTD-phosphoHSP20 peptide alone is sufficient to inhibit force maintenance and likely has a direct effect on the target of phosphorylated HSP20. These results suggest that transduction of phosphopeptide analogs of HSP20 directly alters physiological responses of intact muscles. The data also support a direct role for phosphorylated HSP20 in mediating vasorelaxation. PMID- 12738804 TI - Microtubular interactions of presenilin direct kinesis of Abeta peptide and its precursors. AB - In our previous study we demonstrated that presenilin 1 (PS1) interacts with cytoplasmic linker protein 170/Restin (CLIP-170/Restin). Herein we show that disruption of the interaction of these proteins within neuronal cell-lines (SY5Y and N2a) can be accomplished by the transfection of vectors that drive the expression of peptide fragments corresponding to their binding domains (BDPs). Interestingly, the disruption of the PS1/CLIP-170 complex is associated with both decreased secretion of endogenous Abeta and decreased uptake of exogenous Abeta from the medium. BDP-expressing cells were also more resistant to surges of Abeta secretion induced by thapsigargin and ionomycin (that elevate intracellular calcium concentrations) and mutations in PS1 linked to familial Alzheimer's disease. Uptake of Abeta by SY5Y cells was amplified when preincubated with ApoE and was mediated through lipoprotein receptor-related protein (LRP). BDP expressing cells or cells treated with PS1 anti-sense oligonucleotides were less capable of taking up Abeta from the medium compared with controls, indicating that the PS1/CLIP-170 interaction is involved and that PS1 cannot be substituted. In this study, we also mapped the minimum binding domains (mBDPs) of PS1 and CLIP 70 to regions corresponding to the N-terminal end of the large cytoplasmic loop of PS1 and the metal binding motif-containing C-terminal end of CLIP-170. Further, our data obtained from experiments involving in vitro taxol polymerization of tubulin and confocal immunofluorescence suggest that PS1, via CLIP-170, may serve as an anchor to the microtubules for specific subcellular fractions containing amyloidogenic fragments. Interestingly, Notch is absent from this population of microtubule binding subcellular fractions and its cleavage was unaffected in cells transfected with the PS1-based BDP. This raises the possibility that the interaction of PS1 with CLIP-170 could provide the conceptual basis for anti-amyloidogenic therapeutic strategies with improved specificity. However, this approach may be hampered by a low efficiency, because it may also block Abeta clearance from the interstitial space of the CNS. PMID- 12738806 TI - Identification of two additional members of the membrane-bound dipeptidase family. AB - We have cloned two mouse cDNAs encoding previously unidentified membrane-bound dipeptidases [membrane-bound dipeptidase-2 (MBD-2) and membrane-bound dipeptidase 3 (MBD-3)] from membrane-bound dipeptidase-1 (MBD-1) deficient mice (Habib, G.M., Shi, Z-Z., Cuevas, A.A., Guo, Q., Matzuk, M.M., and Lieberman, M.W. (1998) Proc. Natl. Acad. Sci. USA 95, 4859-4863). These enzymes are closely related to MBD-1 (EC 3.4.13.19), which is known to cleave leukotriene D4 (LTD4) and cystinyl-bis glycine. MBD-2 cDNA is 56% identical to MBD-1 with a predicted amino acid identity of 33%. The MBD-3 and MBD-1 cDNAs share a 55% nucleotide identity and a 39% predicted amino acid sequence identity. All three genes are tightly linked on the same chromosome. Expression of MBD-2 and MBD-3 in Cos cells indicated that both are membrane-bound through a glycosylphosphatidyl-inositol linkage. MBD-2 cleaves leukotriene D4 (LTD4) but not cystinyl-bis-glycine, while MBD-3 cleaves cystinyl-bis-glycine but not LTD4. MBD-1 is expressed at highest levels in kidney, lung, and heart and is absent in spleen, while MBD-2 is expressed at highest levels in lung, heart, and testis and at somewhat lower levels in spleen. Of the tissues examined, MBD-3 expression was detected only in testis. Our identification of a second enzyme capable of cleaving LTD4 raises the possibility that clearance of LTD4 during asthma and in related inflammatory conditions may be mediated by more than one enzyme. PMID- 12738807 TI - Hydrogen sulfide induces serum-independent cell cycle entry in nontransformed rat intestinal epithelial cells. AB - Hydrogen sulfide (H2S), produced by commensal sulfate-reducing bacteria, is an environmental insult that potentially contributes to chronic intestinal epithelial disorders. We tested the hypothesis that exposure of nontransformed intestinal epithelial cells (IEC-18) to the reducing agent sodium hydrogen sulfide (NaHS) activates molecular pathways that underlie epithelial hyperplasia, a phenotype common to both ulcerative colitis (UC) and colorectal cancer. Exposure of IEC-18 cells to NaHS rapidly increased the NADPH/NADP ratio, reduced the intracellular redox environment, and inhibited mitochondrial respiratory activity. The addition of 0.2-5 mM NaHS for 4 h increased the IEC-18 proliferative cell fraction (P<0.05), as evidenced by analysis of the cell cycle and proliferating cell nuclear antigen expression, while apoptosis occurred only at the highest concentration of NaHS. Thirty minutes of NaHS exposure increased (P<0.05) c-Jun mRNA concentrations, consistent with the observed activation of mitogen activated protein kinases (MAPK). Microarray analysis confirmed an increase (P<0.05) in MAPK-mediated proliferative activity, likely reflecting the reduced redox environment of NaHS-treated cells. These data identify functional pathways by which H2S may initiate epithelial dysregulation and thereby contribute to UC or colorectal cancer. Thus, it becomes crucial to understand how genetic background may affect epithelial responsiveness to this bacterial-derived environmental insult. PMID- 12738808 TI - Caudal dysgenesis in Islet-1 transgenic mice. AB - Maternal diabetes during pregnancy is responsible for the occurrence of diabetic embryopathy, a spectrum of birth defects that includes heart abnormalities, neural tube defects, and caudal dysgenesis syndromes. Here, we report that mice transgenic for the homeodomain transcription factor Isl-1 develop profound caudal growth defects that resemble human sacral/caudal agenesis. Isl-1 is normally expressed in the pancreas and is required for pancreas development and endocrine cell differentiation. Aberrant regulation of this pancreatic transcription factor causes increased mesodermal cell death, and the severity of defects is dependent on transgene dosage. Together with the finding that mutation of the pancreatic transcription factor HLXB9 causes sacral agenesis, our results implicate pancreatic transcription factors in the pathogenesis of birth defects associated with diabetes. PMID- 12738809 TI - Pericellular plasmin induces smooth muscle cell anoikis. AB - Smooth muscle cell (SMC) rarefaction is involved in the development of several vascular pathologies. We suggest that the plasminogen activation system is a potential extracellular signal that can induce pericellular proteolysis and apoptosis of vascular SMCs. Using primary cultures of arterial SMCs, we show that plasmin generated from plasminogen on the cell surface induces cell retraction and fibronectin fragmentation, leading to detachment and morphological/biochemical changes characteristic of apoptosis (also called anoikis). The generation of cell-bound plasmin mediated by tissue-type plasminogen activator (t-PA), constitutively expressed by VSMCs, requires binding of plasminogen to the cell surface and is inhibited by epsilon-aminocaproic acid (IC50=0.9+/-0.2 mM), a competitor of plasminogen binding to membrane glycoproteins. Conversely, addition of alpha2-antiplasmin, which blocks free plasmin in the cell supernatant, could not fully prevent anoikis. Finally, an MMP inhibitor failed to prevent VSMC anoikis, arguing for a direct involvement of plasmin in this phenomenon. Indeed, similar changes are induced by plasmin directly added to VSMCs or to arterial rings, ex-vivo. We show for the first time that pathological anoikis can be triggered by a process that requires functional assembly of the plasminogen activation system on the surface of VSMCs. PMID- 12738810 TI - Chronic hyperglycemia impairs insulin secretion by affecting insulin receptor expression, splicing, and signaling in RIN beta cell line and human islets of Langerhans. AB - Recent evidence suggests that insulin signaling through the insulin receptor A type (Ex11-), regulates insulin gene transcription. Because chronic hyperglycemia negatively affects insulin receptor function and regulates alternative splicing of the insulin receptor, we inquired whether chronic exposure of pancreatic beta cells to high glucose results in alterations in insulin signaling due to changes in insulin receptor expression and relative abundance of its spliced isoforms. Our results demonstrate that the insulin receptor is localized in insulin secretory vescicles in human pancreatic beta-cells. Furthermore, we find that alterations in insulin expression and secretion caused by chronic exposure to high glucose are paralleled by decreased insulin receptor expression and increased relative abundance of the Ex11+ isoform in both human islets and RIN beta-cells. PDX-1 and HMGI(Y) transcription factors are down-regulated by high glucose. These changes are associated with defects in insulin signaling involving insulin receptor-associated PI 3-kinase/Akt/PHAS-I pathway in RIN beta-cells. Re expression in RIN beta-cells chronically exposed to high glucose of the Ex11-, but not the Ex11+, isoform restored insulin mRNA expression. These data suggest that changes in early steps of insulin receptor signaling may play a role in determining beta-cell dysfunction caused by chronic hyperglycemia. PMID- 12738811 TI - A selective inhibitor of inducible nitric oxide synthase inhibits exhaled breath nitric oxide in healthy volunteers and asthmatics. AB - The inducible isoenzyme of nitric oxide synthase (iNOS) generates nitric oxide (NO) in inflammatory diseases such as asthma. The prodrug L-N6-(1 iminoethyl)lysine 5-tetrazole amide (SC-51) is rapidly converted in vivo to the active metabolite L-N6-(1-iminoethyl)lysine (L-NIL). Initially, we performed in vitro experiments in human primary airway epithelial cells to demonstrate that L NIL causes inhibition of iNOS. In a randomized double-blind placebo-controlled crossover trial, SC-51 was administered as a single oral dose (20 or 200 mg) in separate cohorts of healthy volunteers (two groups of n=12) and mild asthmatic patients (two groups of n=12). SC-51 (200 mg) reduced exhaled breath NO levels to <2 ppb in both healthy volunteers (P<0.001) and mild asthmatics (P<0.001) within 15 min, representing >90% inhibition of baseline levels of NO in asthmatic patients, with the effects lasting at least 72 h. There were no significant effects on blood pressure, pulse rate, or respiratory function (FEV1). This study demonstrates that an inhibitor of iNOS produces marked inhibition of exhaled breath NO in normal and asthmatic subjects without producing the side effects observed following the systemic administration of non-selective NOS inhibitors, and thus provides support for the potential use of iNOS inhibitors to treat a range of inflammatory clinical disorders. PMID- 12738812 TI - Interleukin-8 secreted by endothelial cells induces chemotaxis of melanoma cells through the chemokine receptor CXCR1. AB - There is increasing evidence that both cell adhesion molecules and soluble factors are involved in tumor metastasis. We have found that endothelial cells secrete chemoattractants that can induce melanoma cell chemotaxis. Protein separation on an ion-exchange column shows the association of IL-8 with fractions that contain the chemoattractant activity. This activity is completely lost from the conditioned medium after immunoprecipitation with anti-IL-8 antibodies, indicating that IL-8 is the major melanoma chemoattractant secreted by endothelial cells. IL-877, the predominant endothelial IL-8 isoform that contains 77 amino acids, is found to be twice as potent as the more common 72-amino acid isoform IL-872. Antibody inhibition studies indicate that the chemotactic response of melanoma cells is mediated by the CXC-chemokine receptor CXCR1 and not by the more promiscuous CXCR2. When stimulated by tumor necrosis factor alpha, the nonresponsive WM35 melanoma cells synthesize a higher level of CXCR1 and become chemotactic toward interleukin (IL)-8. Pretreatment of cells with pertussis toxin nullifies their chemotactic response, suggesting the involvement of G proteins. Antibodies against either IL-8 or CXCR1 inhibit melanoma transendothelial migration in a coculture assay by 30%. These results are consistent with a role for IL-8-induced chemotaxis in the transendothelial migration of melanoma cells. PMID- 12738814 TI - Interaction of discoidin domain receptor 1 isoform b (DDR1b) with collagen activates p38 mitogen-activated protein kinase and promotes differentiation of macrophages. AB - Discoidin domain receptor 1 (DDR1) is a receptor tyrosine kinase activated by collagen. DDR1 is constitutively expressed in a variety of normal and transformed epithelial cells and plays a role in cell migration and differentiation through as yet unidentified signaling pathways. We previously reported inducible expression of DDR1 in human leukocytes and suggested a role for the DDR1a isoform in leukocyte migration through extracellular matrix. Here, we evaluated the contribution of DDR1 in the differentiation of the human monocytic THP-1 cells overexpressing these isoforms and of primary macrophages. Interestingly, collagen activation of DDR1b, but not DDR1a, further promoted phorbol ester-induced differentiation of THP-1 cells as determined by reduced cell proliferation and up regulated expression of HLA-DR, CD11c, CD14, and CD40. Collagen activation of DDR1b also induced the recruitment and phosphorylation of Shc and subsequent phosphorylation of p38 mitogen-activated protein (MAP) kinase and its substrate ATF2. A p38 MAP kinase inhibitor, SB203580, completely inhibited DDR1b-mediated HLA-DR expression. Activation of DDR1 endogenously expressed on macrophages also up-regulated their HLA-DR expression in a p38 MAP kinase-dependent manner. Thus, DDR1b in response to collagen transduces signals that promote maturation/differentiation of HLA-DR-positive antigen-presenting cells and contributes to the development of adaptive immunity in a tissue microenvironment. PMID- 12738816 TI - Weapons laboratories. DOE breaks with tradition, puts Los Alamos up for bids. PMID- 12738815 TI - Stem cells: still here, still waiting. PMID- 12738813 TI - Impairment of vascular endothelial nitric oxide synthase activity by advanced glycation end products. AB - Endothelial damage is believed to play a key role in the development of both micro- and macrovascular disease in diabetes, and advanced glycation end products (AGEs) may contribute importantly to this. To determine whether glucose-derived AGEs can cause endothelial dysfunction, we examined the effects of albumin AGE modified by glucose (AGE-Glu) both in vivo, after injection into rabbit femoral artery, and in vitro on rabbit aortic rings and cultured human umbilical vein endothelial cells (HUVEC). Exposure of blood vessels to AGE-Glu, in vivo and in vitro, inhibited endothelium-dependent vasorelaxation, whereas unmodified albumin did not. In isolated rabbit aorta, this effect was reversible after AGE-Glu washout, and the response to the endothelium-independent vasodilator sodium nitroprusside was unaffected by AGE-Glu. In HUVEC, AGE-Glu inhibited endothelial nitric oxide synthase activity, and this was associated with a decrease in serine phosphorylation of this enzyme. Longer term (72 h) incubation decreased HUVEC viability. Use of specific antibodies demonstrated that these effects were mediated by N(epsilon)-(carboxymethyl)lysine (CML), an important AGE found in vivo, and by the AGE-R1 receptor. Furthermore, these effects all occurred at CML concentrations similar to those found in the plasma of diabetic patients. These results suggest an important role of AGE in the pathogenesis of diabetic vasculopathy. PMID- 12738817 TI - Anthrax vaccine. NIAID's nasty surprise. PMID- 12738818 TI - National Institutes of Health. A call from downtown to shed staff, authority. PMID- 12738819 TI - Biomedical research. Rubin to head Hughes Institute's new 'farm'. PMID- 12738820 TI - Infectious diseases. Smallpox vaccination campaign in the doldrums. PMID- 12738821 TI - Space physics. NASA orders make-or-break tests for gravity probe. PMID- 12738822 TI - Longevity research. Single signal unites treatments that prolong life. PMID- 12738823 TI - National Academy of Sciences. Twenty-one women in 2003 class. PMID- 12738824 TI - Quantum physics. In clone wars, quantum computers need not apply. PMID- 12738825 TI - Paleobiology. Megafauna died from big kill, not big chill. PMID- 12738827 TI - Military research. Mach 12 by 2012? PMID- 12738826 TI - Infectious diseases. Up close and personal with SARS. PMID- 12738828 TI - Iraqi archaeology. Antiquity experts endorse plans to restore shattered collection. PMID- 12738829 TI - Romania. Unexpected riches from a fabled gold mine. PMID- 12738831 TI - Physical anthropology and paleoanthropology meeting. A shrunken head for African Homo erectus. PMID- 12738830 TI - Physical anthropology and paleoanthropology meeting. A miss for moderns and Neandertals. PMID- 12738832 TI - Physical anthropology and paleoanthropology meeting. First modern remains in Europe. PMID- 12738833 TI - Conservation biology. Rapid evolution can foil even the best-laid plans. PMID- 12738834 TI - Communication from space: radio and optical. PMID- 12738836 TI - Remembering great mentors. PMID- 12738835 TI - Persistent organic pollutants and policy. PMID- 12738838 TI - A random samples clarification. PMID- 12738837 TI - Psychiatric drugs and children. PMID- 12738839 TI - Another random samples clarification. PMID- 12738840 TI - Embryonic stem cells. Stem cell programs. AB - The availability of human embryonic stem cell lines provides an important tool for scientists to explore the fundamental mechanisms that regulate differentiation into specific cell types. When more is known about the mechanisms that govern these processes, human embryonic stem cells may be clinically useful in generating cell types that have been damaged or depleted by a variety of human diseases. The NIH is actively pursuing a variety of initiatives to promote this developing research field, while continuing and expanding its long-standing investment in adult stem cells and research. PMID- 12738841 TI - Stem cells. Setting standards for human embryonic stem cells. PMID- 12738842 TI - Ecology. Cryptic herbivores of the rainforest canopy. PMID- 12738844 TI - Geology. Deep origin of hotspots--the mantle plume model. PMID- 12738843 TI - Biomedicine. A view from the top--prion diseases from 10,000 feet. PMID- 12738845 TI - Geology. Is "hotspot" volcanism a consequence of plate tectonics? PMID- 12738847 TI - Manganese: a high-octane dispute. PMID- 12738848 TI - State Court to rule on manganese fume claims. PMID- 12738849 TI - The interface between the biological and inorganic worlds: iron-sulfur metalloclusters. AB - Complex iron-sulfur metalloclusters form the active sites of the enzymes that catalyze redox transformations of N2, CO, and H2, which are likely components of Earth's primordial atmosphere. Although these centers reflect the organizational principles of simpler iron-sulfur clusters, they exhibit extensive elaborations that confer specific ligand-binding and catalytic properties. These changes were probably achieved through evolutionary processes, including the fusion of small clusters, the addition of new metals, and the development of cluster assembly pathways, driven by selective pressures resulting from changes in the chemical composition of the biosphere. PMID- 12738850 TI - Transition metal speciation in the cell: insights from the chemistry of metal ion receptors. AB - The essential transition metal ions are avidly accumulated by cells, yet they have two faces: They are put to use as required cofactors, but they also can catalyze cytotoxic reactions. Several families of proteins are emerging that control the activity of intracellular metal ions and help confine them to vital roles. These include integral transmembrane transporters, metalloregulatory sensors, and diffusible cytoplasmic metallochaperone proteins that protect and guide metal ions to targets. It is becoming clear that many of these proteins use atypical coordination chemistry to accomplish their unique goals. The different coordination numbers, types of coordinating residues, and solvent accessibilities of these sites are providing insight into the inorganic chemistry of the cytoplasm. PMID- 12738851 TI - Boon and bane of metal ions in medicine. AB - In biological systems metal ions promote responses that range from deficiency to toxicity. Some, such as iron and zinc, have a known optimal intake range for normal, healthy individuals. Metal ions contained within well-designed molecules already constitute a great boon for the medicinal pharmacopoeia. However, whether essential or not, the threshold for toxicity can be very low. One of the challenges of designing metal-based drugs is to balance the potential toxicity of an active formulation with the substantial positive impact of these increasingly common therapeutic and diagnostic aids. PMID- 12738852 TI - The ecology of arsenic. AB - Arsenic is a metalloid whose name conjures up images of murder. Nonetheless, certain prokaryotes use arsenic oxyanions for energy generation, either by oxidizing arsenite or by respiring arsenate. These microbes are phylogenetically diverse and occur in a wide range of habitats. Arsenic cycling may take place in the absence of oxygen and can contribute to organic matter oxidation. In aquifers, these microbial reactions may mobilize arsenic from the solid to the aqueous phase, resulting in contaminated drinking water. Here we review what is known about arsenic-metabolizing bacteria and their potential impact on speciation and mobilization of arsenic in nature. PMID- 12738853 TI - The biogeochemical cycles of trace metals in the oceans. AB - Planktonic uptake of some essential metals results in extraordinarily low concentrations in surface seawater. To sequester or take up these micronutrients, various microorganisms apparently release strong complexing agents and catalyze redox reactions that modify the bioavailability of trace metals and promote their rapid cycling in the upper water column. In turn, the low availability of some metals controls the rate of photosynthesis in parts of the oceans and the transformation and uptake of major nutrients such as nitrogen. The extremely low concentrations of several essential metals are both the cause and the result of ultraefficient uptake systems in the plankton and of widespread replacement of metals by one another for various biochemical functions. PMID- 12738854 TI - Mutational analysis of the tyrosine kinome in colorectal cancers. PMID- 12738855 TI - Salt-pump mechanism for contaminant intrusion into coastal aquifers. PMID- 12738856 TI - Calculated phonon spectra of plutonium at high temperatures. AB - We constructed computer-based simulations of the lattice dynamical properties of plutonium using an electronic structure method, which incorporates correlation effects among the f-shell electrons and calculates phonon spectra at arbitrary wavelengths. Our predicted spectrum for the face-centered cubic delta phase agrees well with experiments in the elastic limit and explains unusually large shear anisotropy of this material. The spectrum of the body-centered cubic phase shows an instability at zero temperature over a broad region of the wave vectors, indicating that this phase is highly anharmonic and can be stabilized at high temperatures by its phonon entropy. PMID- 12738857 TI - Microfluidic memory and control devices. AB - We demonstrate microscopic fluidic control and memory elements through the use of an aqueous viscoelastic polymer solution as a working fluid. By exploiting the fluid's non-Newtonian rheological properties, we were able to demonstrate both a flux stabilizer and a bistable flip-flop memory. These circuit elements are analogous to their solid-state electronic counterparts and could be used as components of control systems for integrated microfluidic devices. Such miniaturized fluidic circuits are insensitive to electromagnetic interference and may also find medical applications for implanted drug-delivery devices. PMID- 12738859 TI - Sediment-dispersed extraterrestrial chromite traces a major asteroid disruption event. AB - Abundant extraterrestrial chromite grains from decomposed meteorites occur in middle Ordovician (480 million years ago) marine limestone over an area of approximately 250,000 square kilometers in southern Sweden. The chromite anomaly gives support for an increase of two orders of magnitude in the influx of meteorites to Earth during the mid-Ordovician, as previously indicated by fossil meteorites. Extraterrestrial chromite grains in mid-Ordovician limestone can be used to constrain in detail the temporal variations in flux of extraterrestrial matter after one of the largest asteroid disruption events in the asteroid belt in late solar-system history. PMID- 12738858 TI - A mesoscale iron enrichment in the western subarctic Pacific induces a large centric diatom bloom. AB - We have performed an in situ test of the iron limitation hypothesis in the subarctic North Pacific Ocean. A single enrichment of dissolved iron caused a large increase in phytoplankton standing stock and decreases in macronutrients and dissolved carbon dioxide. The dominant phytoplankton species shifted after the iron addition from pennate diatoms to a centric diatom, Chaetoceros debilis, that showed a very high growth rate, 2.6 doublings per day. We conclude that the bioavailability of iron regulates the magnitude of the phytoplankton biomass and the key phytoplankton species that determine the biogeochemical sensitivity to iron supply of high-nitrate, low-chlorophyll waters. PMID- 12738860 TI - Efficient epoxidation of olefins with >/=99% selectivity and use of hydrogen peroxide. AB - Epoxides are an important class of industrial chemicals that have been used as chemical intermediates. Catalytic epoxidation of olefins affords an interesting production technology. We found a widely usable green route to the production of epoxides: A silicotungstate compound, [gamma-SiW10O34(H2O)2]4-, is synthesized by protonation of a divacant, lacunary, Keggin-type polyoxometalate of [gamma SiW10O36]8- and exhibits high catalytic performance for the epoxidation of various olefins, including propylene, with a hydrogen peroxide (H2O2) oxidant at 305 kelvin. The effectiveness of this catalyst is evidenced by >/=99% selectivity to epoxide, >/=99% efficiency of H2O2 utilization, high stereospecificity, and easy recovery of the catalyst from the homogeneous reaction mixture. PMID- 12738861 TI - State-specific correlation of coincident product pairs in the F + CD4 reaction. AB - When a chemical reaction forms two molecular products, even if the state-resolved differential cross section (DCS) for each product is obtained individually, the coincident attributes of the coproducts are still lacking. We exploit a method that provides coincidence information by measuring the state-resolved, pair correlated DCS. Exemplified by the reaction F + CD4 --> DF + CD3, a time-sliced ion velocity imaging technique was used to measure the velocity distribution of a state-selected CD3 product and to reveal the information of the coincident DF in a state-correlated manner. The correlation of different product state pairs shows a striking difference, which opens up a new way to unravel the complexity of a polyatomic reaction. PMID- 12738862 TI - Explaining the abundance of ants in lowland tropical rainforest canopies. AB - The extraordinary abundance of ants in tropical rainforest canopies has led to speculation that numerous arboreal ant taxa feed principally as "herbivores" of plant and insect exudates. Based on nitrogen (N) isotope ratios of plants, known herbivores, arthropod predators, and ants from Amazonia and Borneo, we find that many arboreal ant species obtain little N through predation and scavenging. Microsymbionts of ants and their hemipteran trophobionts might play key roles in the nutrition of taxa specializing on N-poor exudates. For plants, the combined costs of biotic defenses and herbivory by ants and tended Hemiptera are substantial, and forest losses to insect herbivores vastly exceed current estimates. PMID- 12738863 TI - Long-term effects of wildfire on ecosystem properties across an island area gradient. AB - Boreal forest soils play an important role in the global carbon cycle by functioning as a large terrestrial carbon sink or source, and the alteration of fire regime through global change phenomena may influence this role. We studied a system of forested lake islands in the boreal zone of Sweden for which fire frequency increases with increasing island size. Large islands supported higher plant productivity and litter decomposition rates than did smaller ones, and, with increasing time since fire, litter decomposition rates were suppressed sooner than was ecosystem productivity. This contributes to greater carbon storage with increasing time since fire; for every century without a major fire, an additional 0.5 kilograms per square meter of carbon becomes stored in the humus. PMID- 12738864 TI - Structural basis of multiple drug-binding capacity of the AcrB multidrug efflux pump. AB - Multidrug efflux pumps cause serious problems in cancer chemotherapy and treatment of bacterial infections. Yet high-resolution structures of ligand transporter complexes have previously been unavailable. We obtained x-ray crystallographic structures of the trimeric AcrB pump from Escherichia coli with four structurally diverse ligands. The structures show that three molecules of ligands bind simultaneously to the extremely large central cavity of 5000 cubic angstroms, primarily by hydrophobic, aromatic stacking and van der Waals interactions. Each ligand uses a slightly different subset of AcrB residues for binding. The bound ligand molecules often interact with each other, stabilizing the binding. PMID- 12738865 TI - Molecular architecture of the multiprotein splicing factor SF3b. AB - The splicing factor SF3b is a multiprotein complex essential for the accurate excision of introns from pre-messenger RNA. As an integral component of the U2 small nuclear ribonucleoprotein (snRNP) and the U11/U12 di-snRNP, SF3b is involved in the recognition of the pre-messenger RNA's branch site within the major and minor spliceosomes. We have determined the three-dimensional structure of the human SF3b complex by single-particle electron cryomicroscopy at a resolution of less than 10 angstroms, allowing identification of protein domains with known structural folds. The best fit of a modeled RNA-recognition motif indicates that the protein p14 is located in the central cavity of the complex. The 22 tandem helical repeats of the protein SF3b155 are located in the outer shell of the complex enclosing p14. PMID- 12738866 TI - Recombinant antibodies to the small GTPase Rab6 as conformation sensors. AB - Here we report an approach, based on antibody phage display, to generate molecular conformation sensors. Recombinant antibodies specific to the guanosine triphosphate (GTP)-bound conformation of the small guanosine triphosphatase (GTPase) Rab6, a regulator of membrane traffic, were generated and used to locate Rab6.GTP in fixed cells, and, after green fluorescent protein (GFP) tagging and intracellular expression, to follow Rab6.GTP in vivo. Rab6 was in its GTP-bound conformation on the Golgi apparatus and transport intermediates, and the geometry of transport intermediates was modulated by Rab6 activity. More generally, the same approach could be applied to other molecules that can be locked in a particular conformation in vitro. PMID- 12738867 TI - Unmasking the S-phase-promoting potential of cyclin B1. AB - In higher eukaryotes, the S phase and M phase of the cell cycle are triggered by different cyclin-dependent kinases (CDKs). For example, in frog egg extracts, Cdk1-cyclin B catalyzes entry into mitosis but cannot trigger DNA replication. Two hypotheses can explain this observation: Either Cdk1-cyclin B fails to recognize the key substrates of its S-phase-promoting counterparts, or its activity is somehow regulated to prevent it from activating DNA synthesis. Here, we show that Cdk1-cyclin B1 has cryptic S-phase-promoting abilities that can be unmasked by relocating it from the cytoplasm to the nucleus and moderately stimulating its activity. Subcellular localization of vertebrate CDKs and the control of their activity are thus critical factors for determining their specificity. PMID- 12738868 TI - Asymmetrical allocation of NMDA receptor epsilon2 subunits in hippocampal circuitry. AB - Despite its implications for higher order functions of the brain, little is currently known about the molecular basis of left-right asymmetry of the brain. Here we report that synaptic distribution of N-methyl-D-aspartate (NMDA) receptor GluRepsilon2 (NR2B) subunits in the adult mouse hippocampus is asymmetrical between the left and right and between the apical and basal dendrites of single neurons. These asymmetrical allocations of epsilon2 subunits differentiate the properties of NMDA receptors and synaptic plasticity between the left and right hippocampus. These results provide a molecular basis for the structural and functional asymmetry of the mature brain. PMID- 12738869 TI - Eye-specific retinogeniculate segregation independent of normal neuronal activity. AB - The segregation of initially intermingled left and right eye inputs to the dorsal lateral geniculate nucleus (DLGN) during development is thought to be in response to precise spatial and temporal patterns of spontaneous ganglion cell activity. To test this hypothesis, we disrupted the correlated activity of neighboring ganglion cells in the developing ferret retina through immunotoxin depletion of starburst amacrine cells. Despite the absence of this type of correlated activity, left and right eye inputs segregated normally in the DLGN. By contrast, when all spontaneous activity was blocked, the projections from the two eyes remained intermingled. Thus, certain features of normal neural activity patterns are not required for the formation of eye-specific projections to the DLGN. PMID- 12738870 TI - Episodic tremor and slip on the Cascadia subduction zone: the chatter of silent slip. AB - We found that repeated slow slip events observed on the deeper interface of the northern Cascadia subduction zone, which were at first thought to be silent, have unique nonearthquake seismic signatures. Tremorlike seismic signals were found to correlate temporally and spatially with slip events identified from crustal motion data spanning the past 6 years. During the period between slips, tremor activity is minor or nonexistent. We call this associated tremor and slip phenomenon episodic tremor and slip (ETS) and propose that ETS activity can be used as a real-time indicator of stress loading of the Cascadia megathrust earthquake zone. PMID- 12738872 TI - Radio-frequency spectroscopy of ultracold fermions. AB - Radio-frequency techniques were used to study ultracold fermions. We observed the absence of mean-field "clock" shifts, the dominant source of systematic error in current atomic clocks based on bosonic atoms. This absence is a direct consequence of fermionic antisymmetry. Resonance shifts proportional to interaction strengths were observed in a three-level system. However, in the strongly interacting regime, these shifts became very small, reflecting the quantum unitarity limit and many-body effects. This insight into an interacting Fermi gas is relevant for the quest to observe superfluidity in this system. PMID- 12738871 TI - Crystal structure of the potassium channel KirBac1.1 in the closed state. AB - The KirBac1.1 channel belongs to the inward-rectifier family of potassium channels. Here we report the structure of the entire prokaryotic Kir channel assembly, in the closed state, refined to a resolution of 3.65 angstroms. We identify the main activation gate and structural elements involved in gating. On the basis of structural evidence presented here, we suggest that gating involves coupling between the intracellular and membrane domains. This further suggests that initiation of gating by membrane or intracellular signals represents different entry points to a common mechanistic pathway. PMID- 12738873 TI - Imperforate anus: US determination of the type with infracoccygeal approach. AB - PURPOSE: To assess the usefulness of infracoccygeal transperineal ultrasonography (US) in differentiation between high- and low-type imperforate anus. MATERIALS AND METHODS: Infracoccygeal US was prospectively performed with a 7-10-MHz linear array transducer prior to corrective surgery in 14 neonates with imperforate anus. The approach site was just inferior to the coccyx and posterior to the anus. Transverse images of the anorectal area were obtained. The puborectalis muscle was identified, and the relationship between the puborectalis muscle and the distal rectal pouch was evaluated. US findings were compared with surgical findings. RESULTS: In 10 neonates, a low-type imperforate anus was correctly diagnosed at infracoccygeal US. In those with low-type imperforate anus, the puborectalis muscle was seen as a hypoechoic U-shaped band (n = 10), and the distal rectal pouch passed through the puborectalis muscle (n = 10). In four neonates with high-type imperforate anus, the puborectalis muscle was not identified (n = 4). CONCLUSION: Infracoccygeal transperineal US enables the determination of the type of imperforate anus. PMID- 12738874 TI - Imaging of vascular gene therapy. AB - Gene therapy is an exciting frontier in medicine today. Many genes have been shown to be useful for treatment of various vascular diseases, including chronic cardiac and limb ischemia syndromes, vasculoproliferative disorder, hypercholesterolemia, atherosclerosis, thrombosis, and hypertension. Precise delivery of genes into target vessels, efficient transfer of genes into vascular cells of the target, and prompt assessment of gene expression over time are three challenging tasks for successful vascular gene therapy. Thus, in vivo imaging methods that can be used to monitor gene delivery and localize gene expression are needed. Modern imaging techniques provide an opportunity to monitor and direct vascular gene therapy. Radiologists play a key role not only in developing and mastering endovascular genetic interventions but also in assessing the success of vascular gene therapy and directing further refinement of vascular gene therapy technology. This article provides an overview of the current status of imaging of vascular gene therapy. PMID- 12738875 TI - When is a diagnosis of sclerosing adenosis acceptable at core biopsy? AB - PURPOSE: To determine concordance of imaging findings and diagnosis of sclerosing adenosis at histopathologic core biopsy and to establish the accuracy of core biopsy when cancer was coexistent. MATERIALS AND METHODS: From a database of 1,166 percutaneous biopsies in which sclerosing adenosis was reported, 88 (7.5%) lesions were identified, and imaging and histopathologic findings were reviewed for concordance. Sclerosing adenosis proved to be a minor component at core biopsy for 44 lesions, including one invasive ductal carcinoma, one ductal carcinoma in situ (DCIS), one focus of atypical ductal hyperplasia (ADH), and one atypical lobular hyperplasia. Sclerosing adenosis was a major (> or =50%) component for 44 lesions, including four malignancies, all DCIS manifested as clustered calcifications (pleomorphic [n = 2] or amorphous [n = 2]), and seven foci of ADH manifested as amorphous calcifications. In 30 patients with 33 lesions without atypia or malignancy, sclerosing adenosis was the major finding at core biopsy (21 lesions at 14-gauge core biopsy and 12 at 11-gauge vacuum assisted biopsy); these patients formed the study population. Mammographic (33 lesions) and sonographic (18 lesions) features were recorded. Twenty-seven lesions had at least 20-month follow-up (n = 25) or excision (n = 2). RESULTS: One spiculated mass was considered discordant and was excised, showing a prospectively unrecognized radial sclerosing lesion with several 2-5-mm foci of invasive tubular and lobular carcinoma. Seventeen (53%) of 32 lesions manifested as masses; 10 (59%) were circumscribed, five (29%) were indistinctly marginated (one with punctate calcifications), and two (12%) were partially circumscribed and partially obscured (one with amorphous calcifications). Fifteen (47%) lesions manifested as clustered calcifications; nine (60%) were amorphous and indistinct, four (27%) were pleomorphic, and two (13%) were punctate. Of 27 lesions with acceptable follow-up, 26 (96%) were believed to have been accurately sampled at core biopsy. Of six radial sclerosing lesions associated with the original 88 lesions, only three (50%) were prospectively recognized. CONCLUSION: Sclerosing adenosis is an acceptable result at core biopsy of circumscribed masses and nonpalpable indistinctly marginated masses and for clustered amorphous, pleomorphic, and punctate calcifications. Recognition and reporting of coexistent radial sclerosing lesions is encouraged and may prompt excision. Malignancy can be seen with sclerosing adenosis; core biopsy was accurate in six (86%) of seven coexistent malignancies in this series. PMID- 12738876 TI - Relationship between left biliary duct system and left portal vein: evaluation with three-dimensional portocholangiography. AB - PURPOSE: To investigate the relationship between the left biliary duct system and the left portal vein by using in vivo analysis of the human liver with three dimensional portocholangiography. MATERIALS AND METHODS: Twenty-seven patients underwent helical computed tomography during both arterial portography and cholangiography. Three-dimensional portocholangiograms were reconstructed to evaluate the relationship between the left biliary duct system and the left portal vein, in particular the umbilical portion. RESULTS: In 16 (59%) patients, segment II and III ducts united just above the umbilical portion or laterally to it, and the segment IV duct joined medially to the umbilical portion. In eight (30%) patients, segment III and IV ducts united medially to the umbilical portion and the segment II duct joined at a point close to the hepatic hilum. In three (11%) patients, three ducts from segments II-IV united at a position immediately medial to the umbilical portion. CONCLUSION: Three types of left bile duct anatomy were seen in relation to the left portal vein. PMID- 12738877 TI - Thin-section CT of severe acute respiratory syndrome: evaluation of 73 patients exposed to or with the disease. AB - PURPOSE: To retrospectively analyze the thin-section computed tomographic (CT) features in patients with severe acute respiratory syndrome (SARS) at the authors' institution. MATERIALS AND METHODS: From March 11, 2003, to April 2, 2003, 74 patients with symptoms and signs suggestive of SARS underwent CT of the thorax; all underwent thin-section CT except for one patient who underwent conventional CT. Group 1 (n = 23) patients had symptoms of SARS in keeping with criteria from the Centers for Disease Control and Prevention and a positive chest radiograph. Group 2 (n = 17) patients had a high clinical suspicion of SARS but a normal radiograph. Group 3 (n = 34) patients had minor symptoms and a normal chest radiograph. The thin-section CT images were analyzed for ground-glass opacification or consolidation, lesion size in each lung segment, peripheral or central location, interstitial thickening, and other abnormalities. RESULTS: Thin section CT scans were abnormal only for patients in groups 1 and 2. The patient with only conventional CT scans was in group 3; scans for group 3 patients were normal. Affected segments were predominantly in the lower lobes (91 of 149 affected segments). Common findings included ground-glass opacification, sometimes with consolidation, and interlobular septal and intralobular interstitial thickening. The size of each lesion and the total number of segments involved were smaller in group 2 patients. A majority of patients in group 1 (14 of 23) had mixed central and peripheral lesions. In group 2, however, peripheral lesions were more common (10 of 17). In both groups, a purely central lesion was uncommon (one of 23 in group 1 and two of 17 in group 2). CONCLUSION: Common thin section CT features of SARS are ground-glass opacification and lower lobe and peripheral distribution. PMID- 12738878 TI - Fatty acid synthesis is essential in embryonic development: fatty acid synthase null mutants and most of the heterozygotes die in utero. AB - In animals, including humans, the source of long-chain saturated fatty acids is de novo synthesis, which is mediated by fatty acid synthase (FAS), ingested food, or both. To understand the importance of de novo fatty acid synthesis, we generated FAS knockout mice. The heterozygous FAS mutants (Fasn+/-) are ostensibly normal. In Fasn+/- mice the levels of FAS mRNA and the FAS activity are approximately 50% and 35% lower, respectively, than those of WT mice; hence, FAS levels are affected by gene dosage. When the Fasn+/- mutant mice were interbred, Fasn-/- mice were not produced; thus, FAS is essential during embryonic development. Furthermore, the number of Fasn+/- progeny obtained was 70% less than predicted by Mendelian inheritance, indicating partial haploid insufficiency. Even when one of the parents was WT, the estimated loss of heterozygous progeny was 60%. This loss of Fasn+/- pups appeared to be strain specific and became more pronounced as the heterozygous females produced more litters. Most of the Fasn-/- mutant embryos died before implantation and the Fasn+/- embryos died at various stages of their development. Feeding the breeders a diet rich in saturated fatty acids did not prevent the loss of homoor heterozygotes. These observations are very important in considering teratogenic consequences of drugs aimed at inhibiting FAS activity, to reduce either obesity or the growth of cancerous tissues. PMID- 12738879 TI - Interaction of antimicrobial peptide protegrin with biomembranes. AB - The antimicrobial peptide protegrin-1 (PG-1) interacts with membranes in a manner that strongly depends on membrane lipid composition. In this research we use an approach representing the outer layers of bacterial and red blood cell membranes with lipid monolayers and using a combination of insertion assay, epifluorescence microscopy, and surface x-ray scattering to gain a better understanding of antimicrobial peptide's mechanism of action. We find that PG-1 inserts readily into anionic dipalmitoyl-phosphatidylglycerol, palmitoyl-oleoyl phosphatidylglycerol, and lipid A films, but significantly less so into zwitterionic dipalmitoyl-phosphatidylcholine, palmitoyl-oleoyl phosphatidylcholine, and dipalmitoyl-phosphatidylethanolamine monolayers under similar experimental conditions. Epifluorescence microscopy shows that the insertion of PG-1 into the lipid layer results in the disordering of lipid packing; this disordering effect is corroborated by grazing incidence x-ray diffraction data. X-ray reflectivity measurements further point to the location of the peptide in the lipid matrix. In a pathologically relevant example we show that PG-1 completely destabilizes monolayer composed of lipid A, the major component in the outer membrane of Gram-negative bacteria, which is likely to be the mechanism by which PG-1 disrupts the outer membrane, thus allowing it to reach the target inner membrane. PMID- 12738880 TI - TRAP230/ARC240 and TRAP240/ARC250 Mediator subunits are functionally conserved through evolution. AB - In Saccharomyces cerevisiae Mediator, a subgroup of proteins (Srb8, Srb9, Srb10, and Srb11) form a module, which is involved in negative regulation of transcription. Homologues of Srb10 and Srb11 are found in some mammalian Mediator preparations, whereas no clear homologues have been reported for Srb8 and Srb9. Here, we identify a TRAP240/ARC250 homologue in Schizosaccharomyces pombe and demonstrate that this protein, spTrap240, is stably associated with a larger form of Mediator, which also contains conserved homologues of Srb8, Srb10, and Srb11. We find that spTrap240 and Sch. pombe Srb8 (spSrb8) regulate the same distinct subset of genes and have indistinguishable phenotypic characteristics. Importantly, Mediator containing the spSrb8/spTrap240/spSrb10/spSrb11 subunits is isolated only in free form, devoid of RNA polymerase II. In contrast, Mediator lacking this module associates with the polymerase. Our findings provide experimental evidence for recent suggestions that TRAP230/ARC240 and TRAP240/ARC250 may indeed be the Srb8 and Srb9 homologues of mammalian Mediator. Apparently Srb8/TRAP230/ARC240, Srb9/TRAP240/ARC250, Srb10, and Srb11 constitute a conserved Mediator submodule, which is involved in negative regulation of transcription in all eukaryotes. PMID- 12738881 TI - The putative glutamate receptor 1.1 (AtGLR1.1) functions as a regulator of carbon and nitrogen metabolism in Arabidopsis thaliana. AB - The ability to coordinate carbon (C) and nitrogen (N) metabolism enables plants to regulate development and metabolic responses to different environmental conditions. The regulator(s) or sensor(s) that monitor crosstalk between biosynthetic pathways and ultimately control the flow of C or N through them have remained elusive. We used an antisense strategy to demonstrate that the putative glutamate receptor 1.1 (AtGLR1.1) functions as a regulator of C and N metabolism in Arabidopsis. Seeds from AtGLR1.1-deficient Arabidopsis (antiAtGLR1.1) lines did not germinate in the presence of an animal ionotropic glutamate receptor (iGLR) antagonist, but germination was restored upon coincubation with an iGLR agonist or the putative ligand glutamate. In antiAtGLR1.1 lines, endogenous abscisic acid (ABA) concentrations increased with iGLR antagonist treatments and decreased with coincubation with an iGLR agonist, suggesting that germination was controlled by ABA. antiAtGLR1.1 seedlings also exhibited sensitivity to increased levels of Ca2+ compared with wild type, and they exhibited a conditional phenotype that was sensitive to the C:N ratio. In the presence of C, specifically sucrose, but not glucose, mannitol, or sorbitol, antiAtGLR1.1 seeds did not germinate, but germination was restored upon coincubation with NO3-, but not NH4+. Immunoblot, isoenzyme, and RT-PCR analyses indicate that AtGLR1.1 regulates the accumulation of distinct C- and N-metabolic enzymes, hexokinase 1 (HXK1) and zeaxanthin epoxidase (ABA1), by transcriptional control. We provide a model to describe the role of AtGLR1.1 in C/N metabolism and ABA biosynthesis, which in turn controls seed germination. PMID- 12738882 TI - T lymphoid differentiation in human bone marrow. AB - The unique role of the thymus in the development of T cells was established >4 decades ago. To elucidate how uncommitted lymphoid progenitor cells are instructed to migrate from bone marrow to the thymus to undergo T lymphoid differentiation, we generated and analyzed a genome-wide gene expression profile of CD7+ CD10+ human bone marrow T cell lineage precursors (TLPs) by using the serial analysis of gene expression technique. Unexpectedly, the serial analysis of gene expression profile identified a high number of (pre-) T cell receptor antigen (TCR)-related transcripts in bone marrow TLPs. To determine the configuration of the TCRbeta locus in these cells at a quantitative level, we sorted and analyzed bone marrow TLPs from five donors by single-cell PCR. Similar proportions of TLPs harbored TCRbeta germ-line alleles, D-J, or V-DJ gene rearrangements. Thus, bone marrow TLPs are heterogenous with respect to TCRbeta rearrangement status, suggesting an active recombination machinery that is consistent with the expression of RAG1, RAG2, and TdT in this population. As a hallmark of ongoing TCRbeta V-DJ rearrangement, we could amplify broken-ended recombination-signal sequence DNA intermediates from bone marrow TLPs, but not from mature T cells by ligation-mediated PCR. Approximately half of the TCRbeta rearrangements were compatible with the expression of a functional pre-TCR, which is in agreement with surface expression of pre-Talpha on bone marrow TLPs as shown by confocal laser microscopy and flow cytometry. At a frequency <0.5% of mononucleated cells in human bone marrow, this population is rare, yet it exemplifies T lymphoid differentiation in the human already before immigration into the thymus. PMID- 12738883 TI - Compression forces generated by actin comet tails on lipid vesicles. AB - Polymerizing networks of actin filaments generate force for a variety of movements in living cells, including protrusion of filopodia and lamellipodia, intra- and intercellular motility of certain bacterial and viral pathogens, and motility of endocytic vesicles and other membrane-bound organelles. During actin based motility, coexisting populations of actin filaments exert both pushing and retarding forces on the moving cargo. To examine the distribution and magnitude of forces generated by actin, we have developed a model system where large artificial lipid vesicles coated with the protein ActA from the bacterial pathogen Listeria monocytogenes are propelled by actin polymerization in cytoplasmic extract. We find that motile vesicles associated with actin comet tails are significantly deformed due to an inward compression force exerted by actin polymerization orthogonal to the direction of motion, which is >10-fold greater in magnitude than the component of the force exerted in the direction of motion. Furthermore, there is a spatial segregation of the pushing and retarding forces, such that pushing predominates along the sides of the vesicle, although retarding forces predominate at the rear. We estimate that the total net (pushing minus retarding) force generated by the actin comet tail is approximately 0.4-4 nN. In addition, actin comet tail formation is associated with polarization of the ActA protein on the fluid vesicle surface, which may reinforce the persistence of unidirectional motion by helping to maintain a persistent asymmetry of actin filament density. PMID- 12738884 TI - Coincidence of synteny breakpoints with malignancy-related deletions on human chromosome 3. AB - We have found previously that during tumor growth intact human chromosome 3 transferred into tumor cells regularly looses certain 3p regions, among them the approximately 1.4-Mb common eliminated region 1 (CER1) at 3p21.3. Fluorescence in situ hybridization analysis of 12 mouse orthologous loci revealed that CER1 splits into two segments in mouse and therefore contains a murine/human conservation breakpoint region (CBR). Several breaks occurred in tumors within the region surrounding the CBR, and this sequence has features that characterize unstable chromosomal regions: deletions in yeast artificial chromosome clones, late replication, gene and segment duplications, and pseudogene insertions. Sequence analysis of the entire 3p12-22 revealed that other cancer-associated deletions (regions eliminated from monochromosomal hybrids carrying an intact chromosome 3 during tumor growth and homozygous deletions found in human tumors) colocalized nonrandomly with murine/human CBRs and were characterized by an increased number of local gene duplications and murine/human conservation mismatches (single genes that do not match into the conserved chromosomal segment). The CBR within CER1 contains a simple tandem TATAGA repeat capable of forming a 40-bp-long secondary hairpin-like structure. This repeat is nonrandomly localized within the other tumor-associated deletions and in the vicinity of 3p12 22 CBRs. PMID- 12738885 TI - Molecular basis for the immunostimulatory activity of guanine nucleoside analogs: activation of Toll-like receptor 7. AB - Certain C8-substituted and N7, C8-disubstituted guanine ribonucleosides comprise a class of small molecules with immunostimulatory activity. In a variety of animal models, these agents stimulate both humoral and cellular immune responses. The antiviral actions of these guanosine analogs have been attributed to their ability to induce type I IFNs. However, the molecular mechanisms by which the guanosine analogs potentiate immune responses are not known. Here, we report that several guanosine analogs activate Toll-like receptor 7 (TLR7). 7-Thia-8 oxoguanosine, 7-deazaguanosine, and related guanosine analogs activated mouse immune cells in a manner analogous to known TLR ligands, inducing cytokine production in mouse splenocytes (IL-6 and IL-12, type I and II IFNs), bone marrow derived macrophages (IL-6 and IL-12), and in human peripheral blood leukocytes (type I IFNs, tumor necrosis factor alpha and IL-12). The guanosine congeners also up-regulated costimulatory molecules and MHC I/II in dendritic cells. Genetic complementation studies in human embryonic kidney 293 cells confirmed that the guanosine analogs activate cells exclusively via TLR7. The stimulation of TLR7 by the guanosine analogs in human cells appears to require endosomal maturation because inhibition of this process with chloroquine significantly reduced the downstream activation of NF-kappaB. However, TLR8 activation by R-848 and TLR2 activation by [S-[2,3-bis(palmitoyloxy)-(2-RS)-propyl]-N-palmitoyl-R-Cys S-Ser-Lys4-OH, trihydrochloride)] were not inhibited by chloroquine, whereas TLR9 activation by CpG oligodeoxynucleotides was abolished. In summary, we present evidence that guanosine analogs activate immune cells via TLR7 by a pathway that requires endosomal maturation. Thus, the B cell-stimulating and antiviral activities of the guanosine analogs may be explained by their TLR7-activating capacity. PMID- 12738886 TI - Exo1: a new chemical inhibitor of the exocytic pathway. AB - A phenotypic screen was used to search for drug-like molecules that can interfere with specific steps in membrane traffic. 2-(4-Fluorobenzoylamino)-benzoic acid methyl ester (Exo1), identified in this screen, induces a rapid collapse of the Golgi to the endoplasmic reticulum, thus acutely inhibiting the traffic emanating from the endoplasmic reticulum. Like Brefeldin A (BFA), Exo1 induces the rapid release of ADP-ribosylation factor (ARF) 1 from Golgi membranes but has less effect on the organization of the trans-Golgi network. Our data indicate that Exo1 acts by a different mechanism from BFA. Unlike BFA, Exo1 does not induce the ADP-ribosylation of CtBP/Bars50 and does not interfere with the activity of guanine nucleotide exchange factors specific for Golgi-based ARFs. Thus, Exo1 allows the fatty acid exchange activity of Bars50 to be distinguished from ARF1 activity in the control of Golgi tubulation. PMID- 12738887 TI - Spermatogonial stem cells share some, but not all, phenotypic and functional characteristics with other stem cells. AB - Spermatogonial stem cells (SSCs) are responsible for maintaining spermatogenesis throughout life in the male by continuous production of daughter cells that differentiate into spermatozoa. However, no unique phenotypic markers to identify SSCs have been described. In this study, the SSC surface phenotype was characterized by using flow cytometric cell sorting in conjunction with a transplantation functional assay for SSCs. Highly enriched stem cell activity was found in the MHC class I (MHC-I)-Thy-1+c-kit- cell fraction of the mouse cryptorchid testis. There was little or no stem cell activity in any other fraction. The antigenic phenotype of the MHC-I-Thy-1+c-kit- SSCs was alpha6 integrin+CD24+alphavintegrin-Sca-1-CD34-. Subsequently, testis side population (SP) cells, which are defined by a Hoechst dye efflux assay, were identified. Their surface phenotype was found to be MHC-I+Thy-1-Sca-1+, and the transplantation assay demonstrated that the testis SP and SSCs are distinct populations. In several other tissues, the SP has been shown to contain stem cells, but we found that this characteristic does not define SSCs. The identification of a surface phenotype that allows production of a highly enriched SSC population will facilitate functional and genomic studies and enable further comparison with other stem cells. PMID- 12738889 TI - Trends in stroke and coronary heart disease in the WHO MONICA Project. AB - BACKGROUND AND PURPOSE: Coronary heart disease (CHD) and stroke are leading causes of death and disability. Because they share major common risk factors, it would be expected that trends in mortality and incidence of these 2 major cardiovascular diseases would be similar. METHODS: Data from the World Health Organization (WHO) Multinational Monitoring of Trends and Determinants in Cardiovascular Disease (MONICA) Project were used to compare 10-year trends in mortality, event rates, and case fatality from both CHD and stroke. Fifteen populations in the WHO MONICA Project provided data on both CHD (60 763 events) and stroke (10 442 events) in men and women aged 35 to 64 years (23.4 million person-years of observation in total). RESULTS: Trends for the 2 cardiovascular diseases varied within and between populations, and when data from all populations were combined, trends in CHD and stroke mortality differed in men (P=0.001) but not in women, whereas trends in event rates differed significantly in both men and women (P<0.001 and P=0.011, respectively). The differences in trends for CHD and stroke case fatality were not statistically significant in either men or women. In sensitivity analyses, differences in trends in event rates remained statistically significant in men (P<0.001) but not in women. CONCLUSIONS: Trends for CHD and stroke mortality rates, event rates, and case fatality differ substantially between and within the study populations. PMID- 12738888 TI - Instability of GGL domain-containing RGS proteins in mice lacking the G protein beta-subunit Gbeta5. AB - RGS (regulator of G protein signaling) proteins containing the G protein gamma like (GGL) domain (RGS6, RGS7, RGS9, and RGS11) interact with the fifth member of the G protein beta-subunit family, Gbeta5. This interaction is necessary for the stability of both the RGS protein and for Gbeta5. Consistent with this notion, we have found that elevation of RGS9-1 mRNA levels by transgene expression does not increase RGS9-1 protein level in the retina, suggesting that Gbeta5 levels may be limiting. To examine further the interactions of Gbeta5 and the GGL domain containing RGS proteins, we inactivated the Gbeta5 gene. We found that the levels of GGL domain-containing RGS proteins in retinas and in striatum are eliminated or reduced drastically, whereas the levels of Ggamma2 and RGS4 proteins remain normal in the absence of Gbeta5. The homozygous Gbeta5 knockout (Gbeta5-/-) mice derived from heterozygous knockout mating are runty and exhibit a high preweaning mortality rate. We concluded that complex formation between GGL domain-containing RGS proteins and the Gbeta5 protein is necessary to maintain their mutual stability in vivo. Furthermore, in the absence of Gbeta5 and all four RGS proteins that form protein complexes with Gbeta5, the animals that survive into adulthood are viable and have no gross defects in brain or retinal morphology. PMID- 12738890 TI - Low vitamin B6 but not homocyst(e)ine is associated with increased risk of stroke and transient ischemic attack in the era of folic acid grain fortification. AB - BACKGROUND AND PURPOSE: The introduction of cereal grain folic acid fortification in 1998 has reduced homocyst(e)ine (tHcy) concentrations in the US population. We performed a case-control study to determine the risk of stroke and transient ischemic attack (TIA) associated with tHcy and low vitamin status in a postfortification US sample. METHODS: Consecutive cases with new ischemic stroke/TIA were compared with matched controls. Fasting tHcy, folate, pyridoxal 5'-phosphate (PLP), B12, and MTHFR 677C-->T genotype were measured. RESULTS: Mean PLP was significantly lower in cases than controls (39.97 versus 84.1 nmol/L, P<0.0001). After stroke risk factors were controlled for, a strong independent association was present between stroke/TIA and low PLP (adjusted odds ratio [OR], 4.6; 95% CI, 1.4 to 15.1; P<0.001) but not elevated tHcy (OR, 0.92; 95% CI, 0.4 to 2.1). CONCLUSIONS: Low B6 but not tHcy was strongly associated with cerebrovascular disease in this postfortification, folate-replete sample. PMID- 12738891 TI - Community-based thrombolytic therapy of acute ischemic stroke in Helsinki. AB - BACKGROUND AND PURPOSE: Thrombolysis with alteplase is used in acute ischemic stroke within 3 hours after symptom onset in many stroke centers, but experience remains limited in Europe. METHODS: Using eligibility and management criteria similar to those published by the American Heart Association, we treated 75 consecutive patients aged 21 to 83 years (mean age, 63.6 years; median Scandinavian Stroke Scale score, 32/58) with hemispheric infarction with alteplase in 1998-2001. Their neuroradiological findings (ischemic and hemorrhagic changes) and functional outcome at 3 months were evaluated. RESULTS: Sixty-one percent of the patients had recovered functional independence (Barthel Index 95 to 100) at the 3-month follow-up. On the modified Rankin Scale (mRS), 37% (28/75) of patients had no or minimal symptoms (mRS 0 to 1), while 17% (13/75) remained dependent (mRS 4 to 5) and 5% (4/75) died. Cerebral parenchymal hematomas occurred in 8% (6/75) and hemorrhagic transformation in 8% (6/75) of the patients. Low initial diastolic blood pressure and administration of intravenous antihypertensive medication were associated with unfavorable outcome (mRS 3 to 6). CONCLUSIONS: We conclude that our management protocol for thrombolytic therapy is safe. These rates of functional outcome, case fatality, and hemorrhagic cerebral events compare favorably with those of other published series of stroke thrombolysis with similar time windows and management guidelines. Associations between blood pressure and its treatment during thrombolysis with functional outcome deserve further analysis. PMID- 12738892 TI - Fibroblast growth factor-18 reduced infarct volumes and behavioral deficits after transient occlusion of the middle cerebral artery in rats. AB - BACKGROUND AND PURPOSE: Fibroblast growth factor 18 (FGF18) is expressed in rodent brain and is a trophic factor for neuron-derived cells in culture. The purpose of the present study was to evaluate whether FGF18 was neuroprotective in a rat model of cerebral ischemia and to compare the results with those obtained with FGF2. METHODS: Cerebral ischemia was produced in rats by a transient 2-hour occlusion of the middle cerebral artery (MCAo) with an intraluminal filament followed by 22-hour reperfusion. Starting 15 minutes after MCAo, FGF18 or FGF2 was administered by a 3-hour intravenous infusion. Infarct volumes and behavioral deficits were measured 24 hours after MCAo. RESULTS: Infusion of FGF18 produced dose-dependent reductions in infarct volumes and improvements in tests of reference and working memory, motor ability, and exploratory behavior. FGF18 was more efficacious than FGF2 on virtually all measures examined. The reductions in infarct volume and behavioral deficit were associated with FGF-mediated increases in regional cerebral blood flow. CONCLUSIONS: These results demonstrate that FGF18 is an effective neuroprotective agent in a rat model of transient MCAo. PMID- 12738893 TI - Functional neuroimaging studies of motor recovery after stroke in adults: a review. AB - BACKGROUND: The precise mechanisms of and biological basis for motor recovery after stroke in adults are still largely unknown. Reorganization of the motor system after stroke as assessed by functional neuroimaging is an intriguing but challenging new field of research. Provocative but equivocal findings have been reported to date. SUMMARY OF REVIEW: We present an overview of functional neuroimaging studies (positron emission tomography or functional MRI) of motor tasks in patients recovered or still recovering from motor deficit after stroke. After a brief account of the connectivity of motor systems and the imaging findings in normal subjects, the literature concerning stroke patients is reviewed and discussed, and a general model is proposed. CONCLUSIONS: Both cross sectional and longitudinal studies have demonstrated that the damaged adult brain is able to reorganize to compensate for motor deficits. Rather than a complete substitution of function, the main mechanism underlying recovery of motor abilities involves enhanced activity in preexisting networks, including the disconnected motor cortex in subcortical stroke and the infarct rim after cortical stroke. Involvement of nonmotor and contralesional motor areas has been consistently reported, with the emerging notion that the greater the involvement of the ipsilesional motor network, the better is the recovery. This hypothesis is supported by the enhanced activity of the ipsilesional primary motor cortex induced by motor training and acute pharmacological interventions, in parallel with improved motor function. Further longitudinal studies assessing the relationships between such changes and actual recovery, as well as manipulating such changes by rehabilitation or pharmacological maneuvers, should provide further information on these fundamental questions. This review closes with some perspectives for future research. PMID- 12738894 TI - Cerebral ischemic events after diagnosis of mitral valve prolapse: a community based study of incidence and predictive factors. AB - BACKGROUND AND PURPOSE: Association of mitral valve prolapse (MVP) with ischemic neurological events (INEs) is uncertain. METHODS: In the community of Olmsted County (Minn), we identified all MVP diagnosed (1989 to 1998) in patients in sinus rhythm with no prior history of INE. We measured INE rates and compared them with expected rates in our community to define the excess risk of INE. RESULTS: Among 777 eligible subjects (age, 49+/-20 years; 66% female; follow-up, 5.5+/-3.0 years), 30 patients had at least 1 INE during follow-up (at 10 years, 7+/-1%). Compared with expected INEs in the same community, subjects with MVP showed excess risk of lifetime INE (relative risk [RR], 2.2; 95% CI, 1.5 to 3.2; P<0.001) and during follow-up under purely medical management (RR, 1.8; 95% CI, 1.1 to 2.8; P=0.009). Independent determinants of INE were older age (RR, 1.08 per year; 95% CI, 1.04 to 1.11; P<0.001), mitral thickening (RR, 3.2; 95% CI, 1.4 to 7.4; P=0.008), atrial fibrillation (AFib) during follow-up (RR, 4.3; 95% CI, 1.9 to 10.0; P<0.001), and need for cardiac surgery (RR, 2.5; 95% CI, 1.1 to 5.8; P=0.03). INE 10-year rates were low in patients <50 years of age (0.4+/-0.4%, P=0.60 versus expected) but were excessive in patients >50 years of age (16+/-3%, P<0.001 versus expected) or with thickened leaflets (7+/-2%, P<0.001 versus expected). Predictors of follow-up AFib were age, mitral regurgitation, and left atrium diameter (all P<0.01). CONCLUSIONS: In the community, subjects with MVP display a lifetime excess rate of INE compared with expected. Clinical (older age) and echocardiographic (leaflets thickening) characteristics define patients with MVP at high risk for INE, and subsequent AFib or need for cardiac surgery, both related to the degree of mitral regurgitation, increase the risk of INE. PMID- 12738895 TI - Neuroprotective kappa-opioid receptor agonist BRL 52537 attenuates ischemia evoked nitric oxide production in vivo in rats. AB - BACKGROUND AND PURPOSE: Kappa-opioid receptors (KOR) have been implicated in neuroprotection from ischemic neuronal injury. We tested the effects of a selective and specific KOR agonist, BRL 52537 hydrochloride [(+/-)-1-(3,4 dichlorophenyl)acetyl-2-(1-pyrrolidinyl) methylpiperidine], on infarct volume and nitric oxide production after transient focal ischemia in the rat. METHODS: With the use of the intraluminal filament technique, halothane-anesthetized male Wistar rats (weight, 250 to 300 g) were subjected to 2 hours of focal cerebral ischemia confirmed by Doppler flowmetry. In a blinded randomized fashion, rats were treated with intravenous saline or 1 mg/kg per hour BRL 52537 infusion, initiated 15 minutes before occlusion and maintained until 2 hours of reperfusion. In a second experiment, rats were treated during reperfusion with saline or 1 mg/kg per hour BRL 52537, initiated at onset of reperfusion and continued for 22 hours. In a final experiment, in vivo striatal nitric oxide production was estimated via microdialysis by quantification of citrulline recovery after labeled arginine infusion in striatum of intravenous BRL 52537- or saline-treated rats. RESULTS: In rats treated with BRL 52537 during ischemia and early reperfusion, infarct volume was significantly attenuated in cortex (16+/-6% versus 40+/-7% of ipsilateral cortex in saline group) and in caudoputamen (30+/ 8% versus 66+/-6% of ipsilateral caudoputamen in saline group). Infarct volume was also reduced by treatment administered only during reperfusion in cortex (19+/-8% in BRL 52537 group [n=10] versus 38+/-6% in saline group) and in caudoputamen (35+/-9% versus 66+/-4% in saline group). BRL 52537 treatment markedly attenuated NO production in ischemic striatum compared with saline treated controls. CONCLUSIONS: These data demonstrate that (1) the selective KOR agonist BRL 52537 provides significant neuroprotection from focal cerebral ischemia when given as a pretreatment or as a posttreatment and (2) attenuation of ischemia-evoked nitric oxide production in vivo may represent one mechanism of ischemic neuroprotection. PMID- 12738896 TI - Revisiting the appropriateness of carotid endarterectomy. AB - BACKGROUND AND PURPOSE: In the 1980s, carotid endarterectomy was controversial because proof of efficacy was lacking, complication rates were high, and one third of cases were reported to be inappropriate. Since publication of several randomized controlled trials (RCTs), rates of carotid endarterectomy have doubled nationwide. This study assesses the appropriateness and use of carotid endarterectomy since publication of the RCTs. METHODS: Using the literature, we developed a list of 1557 mutually exclusive indications for carotid endarterectomy and asked a panel of national experts to rate the appropriateness of each indication using the RAND methodology. We used these ratings to assess appropriateness in a sample of 2124 patients who underwent the procedure in 1997 to 1998 in 6 hospitals. We also analyzed the reasons for the procedure and rates of death, stroke, and myocardial infarction within 30 days of surgery. RESULTS: Overall, 84.9% of operations were done for appropriate reasons, 4.5% for uncertain reasons, and 10.6% for inappropriate reasons. Among procedures considered inappropriate, the most common reasons were high comorbidity (46.6%) and minimal stenosis (27.1%). Overall, 72.5% were asymptomatic, 17.4% had a carotid transient ischemic attack, and 10.1% had a stroke. The 30-day rate of death or stroke was 5.47% for symptomatic patients and 2.26% for asymptomatic patients. Among patients having combined carotid and coronary artery bypass graft surgery, the rate was 10.32%. The complication rate in asymptomatic patients with high comorbidity was 5.56%. CONCLUSIONS: Since the RCTs, rates of overuse appear to have fallen considerably, although they are still significant. A major shift has occurred toward operating on asymptomatic patients. Although overall complication rates were low, rates among asymptomatic patients with high comorbidity exceeded recommended thresholds. PMID- 12738897 TI - Does a relationship exist between carotid stenosis and lacunar infarction? AB - BACKGROUND AND PURPOSE: The presence of carotid stenosis (CS) in a patient with lacunar stroke is usually considered an indication of atherosclerosis and not directly related to the development of this infarction subtype. This study was designed to determine the relationship between CS and lacunar infarction (LI) and to assess the differences between single and multiple LIs. METHODS: We classified 330 patients with a first-ever cerebral infarction in the carotid territory into LI and non-LI (NLI) groups. In the LI group, patients with a single LI and those with multiple LIs were identified. In this last subgroup, 2 patterns were identified: 1 subtype with lacunar lesions distributed in both cerebral hemispheres, and another with lesions predominantly in 1 hemisphere. RESULTS: In the LI group, isolated CS was significantly more frequent on the homolateral side than on the contralateral side (odds ratio [OR], 5.5; 95% CI, 1.2 to 23; P=0.03). A significant relationship between the pattern of distribution of the infarctions in only 1 hemisphere and homolateral CS >70% was observed (OR, 4.4; 95% CI, 0.9 to 19; P=0.03). In a multivariate analysis, the following variables were found to predict unilateral multiple LI: left ventricular hypertrophy (OR, 9.1; 95% CI, 2.5 to 33.6) and homolateral CS >75% (OR, 14.4; 95% CI, 2.0 to 99.6). CONCLUSIONS: The significant incidence of isolated ipsilateral CS in patients with LI located in the carotid territory and the relationship of CS to ipsilateral multiple LI suggest that CS has a very important role in the development of LI. PMID- 12738898 TI - Copper-zinc superoxide dismutase affects Akt activation after transient focal cerebral ischemia in mice. AB - BACKGROUND AND PURPOSE: The serine-threonine kinase Akt is activated by phosphorylation at serine-473. After phosphorylation, activated Akt inactivates BAD or caspase-9 or other apoptogenic components, thereby inhibiting cell death. In this study we examined the relationship between Akt phosphorylation and oxidative stress after transient focal cerebral ischemia (FCI) using copper-zinc superoxide dismutase (SOD1) transgenic (Tg) mice. METHODS: The mice were subjected to 60 minutes of middle cerebral artery occlusion by intraluminal suture blockade followed by 1, 4, and 24 hours of reperfusion. Phospho-Akt expression was examined by immunohistochemistry and Western blot analysis. Production of superoxide anion was assessed by the hydroethidine method in both wild-type mice and SOD1 Tg mice. DNA fragmentation was evaluated by terminal deoxynucleotidyl transferase-mediated uridine 5'-triphosphate-biotin nick end labeling (TUNEL). RESULTS: Immunohistochemistry demonstrated that phospho-Akt was constitutively expressed and was decreased in the ischemic core as early as 1 hour after reperfusion, whereas it was temporally increased in the cortex at 4 hours. Phospho-Akt expression was enhanced in the SOD1 Tg mice. Western blot analysis showed that phospho-Akt was maximized 4 hours after reperfusion in the wild-type mice, whereas phospho-Akt was increased as early as 1 hour after ischemia in the SOD1 Tg mice. There was a significant decrease in TUNEL-positive cells in the SOD1 Tg mice compared with the wild-type mice. CONCLUSIONS: The present study suggests that SOD1 may contribute to the early activation of the Akt cell survival signaling pathway and may attenuate subsequent DNA damage after transient FCI. PMID- 12738900 TI - Editorial comment: Salting the brain to improve CBF in SAH patients. PMID- 12738899 TI - Perfusion-weighted magnetic resonance imaging thresholds identifying core, irreversibly infarcted tissue. AB - BACKGROUND AND PURPOSE: Identifying core, irreversibly infarcted tissue and salvageable penumbral tissue is crucial to informed, physiologically guided decision making regarding thrombolytic and other interventional therapies in acute ischemic stroke. Pretreatment perfusion MRI offers promise as a means to differentiate core from penumbral tissues. METHODS: Diffusion-perfusion MRIs were performed before treatment and on day 7 in patients undergoing successful vessel recanalization with intra-arterial thrombolytic therapy. Perfusion maps of the time to peak of the residue function (Tmax) were generated after deconvolution of an arterial input function. Initial perfusion abnormalities and final infarct regions were outlined by hand. Posttreatment images were coregistered to the pretreatment study. Voxel-by-voxel and volume analyses were performed to identify thresholds of perfusion abnormalities that best predict core, irreversibly infarcted tissue. RESULTS: Fourteen patients (4 men, 10 women) with vessel recanalization were studied. Mean age was 73 years, and median entry National Institutes of Health Stroke Scale score was 12. Mean time from symptom onset to start of intra-arterial infusion was 245 minutes and to recanalization was 338 minutes. With a voxel-by-voxel analysis, Tmax > or =6 and > or =8 seconds (sensitivity, 71% and 53%; specificity, 63% and 80%) correlated most highly with day 7 final infarct. With a volume analysis, Tmax > or =6 and > or =8 seconds (r2=0.704 and r2=0.705) correlated most highly with day 7 final infarct. CONCLUSIONS: Perfusion-weighted imaging measures of ischemia severity accurately differentiate irreversibly injured core from penumbral, salvageable tissue. The best threshold for identifying core infarcted tissue is adjusted Tmax of > or =6 to 8 seconds. PMID- 12738901 TI - [Role of nitric oxide and other endothelium-derived factors]. AB - The endothelial cell layer displays the features of a distributed organ and has a variety of biological functions such as keeping the balance between coagulation and fibrinolysis, expression of adhesion molecules for cells in the immune system, metabolism of noradrenaline and 5-hydroxytryptamine, and conversion of angiotensin I and bradykinin. The endothelium also regulates the underlying smooth muscle layer and vascular tone by release of endothelium-derived relaxing factors such as nitric oxide (NO), prostaglandins, and endothelium-derived hyperpolarizing factor (EDHF) as well as vasoconstricting factors such as endothelin, superoxide (O(2)(-)), and thromboxane. We have reviewed the nature, mechanisms of action, and role of these factors in regulation of vascular tone, with special emphasis on NO. By a process catalyzed by NO synthase, NO and citrulline is formed from the substrates molecular O(2) and L-arginine. The main receptor for NO is guanylyl cyclase leading to formation of smooth muscle cyclic guanosinmonophosphate and relaxation. EDHF is an endothelium-derived factor causing vasorelaxation of the underlying smooth muscle layer by hyperpolarization. The nature of EDHF is still unknown, but several candidates for EDHF have been proposed such as potassium ions, hydrogen peroxide, and epoxyeicosatrienoic acids. Prostaglandins such as prostacyclin and prostaglandin E2 binds to specific receptors followed by increases in cyclic adenosinmonophosphate and vasorelaxation, while contractile prostaglandins constrict vessels by activation of thromboxane and endoperoxidase receptors. Superoxide anions induce contraction of vascular smooth muscles cells by scavenging NO. Endothelin is a potent endothelium-derived contractile factor. The synthesis of endothelin-1 is induced by hypoxia, thrombin, interleukin-1, transforming growth factor-beta1, vasopressin, and catecholamines. Cardiovascular risk factors like age, hypertension, and hyperlipidemia are associated with impaired endothelium-dependent vasodilation either as a consequence of increased inactivation of endothelium-derived vasodilators or increased formation of endothelium-derived contracting factors. This imbalance of endothelium-derived factors plays a role for development of atheroslerosis and ischemic vascular diseases. PMID- 12738902 TI - [Vulvar intraepithelial neoplasia]. AB - In order to improve the timely diagnostics and the efficacy of treatment of vulvar intraepithelial neoplasia, vulvar intraepithelial neoplasia diagnostics criteria and the modern treatment strategies are discussed in this review. We propose the present problems of differentiation of vulvar intraepithelial neoplasia from non-neoplastic epithelial disorders. General concepts of the etiology, pathogenesis, factors of risk, such as: human papilloma virus (HPV), sexually transmitted diseases, human immunodeficiency virus (HIV), smoking are summarized. PMID- 12738903 TI - [Treatment and prophylaxis of deep venous thrombosis with low molecular weight heparins (meta-analysis of clinical trials)]. AB - Deep-vein thrombosis is a relevant problem of today's medicine, because the risk to fall ill with this pathology is 2-5%; it increases in senior age. Direct and indirect acting thrombin inhibitors are used for treatment and prevention of vein thrombosis. Though great efficiency and safety of new anticoagulants (especially factor Xa inhibitors) were proven in clinical studies, unfractionated heparin and low molecular weight heparins are still most widely used in clinical practice. Low molecular weight heparins are obtained by depolymerizing heparin: its molecular weight is being decreased to 3-7 kDa, or 18-20 monosaccharides. Low molecular weight heparins strongly inhibit Xa coagulation factor and faintly - IIa, that's why their anti-Xa/anti-IIa proportion is >1 (unfractionated heparin = 1); enoxaparine and nadroparine show up the highest proportion. The low weight of these heparins determines good pharmacodynamic characteristics: excellent assimilation from subcutaneous layer, long circulation in plasma, infrequent side effects. Due to these characteristics they are convenient, safe and economically worth using (used by subcutaneous injections, prescribed only 1-2 times per day, coagulation control not required, possibility for patient to be treated at home); therefore low molecular weight heparins are more and more often used in treatment of deep-vein thrombosis and also in primary and secondary prevention. They are one of the most efficacious contemporary anticoagulants, which allow to decrease the deep-vein thrombosis treatment and prevention costs. This article presents literature review about low molecular weight heparins, their appliance in treatment and prophylaxis of deep-vein thrombosis. PMID- 12738904 TI - [Influence of preoperative biliary drainage and obstructive jaundice on the early outcome of pancreaticoduodenectomy]. AB - OBJECTIVE: To assess the influence of jaundice and preoperative biliary drainage (PBD) on postoperative outcome following pancreaticoduodenectomy (PD). MATERIAL AND METHODS: Data of 122 patients who underwent PD for periampular tumors and pancreatic adenocarcinoma in Kaunas University of Medicine Hospital was analyzed. Patients were divided into groups: those who have not underwent PBD, but had preoperative bilirubin level <50 micromol/l (n=42), patients without PBD, but preoperative bilirubin exceeding 50 micromol/l (n=61) and patients who have undergone preoperative PBD (n=19). RESULTS: Overall morbidity following PD was 55.6%. There were 10.7% pancreatic fistulas, 13.1% septic complications (either intraabdominal sepsis or wound infection) and 9% of postoperative intraabdominal bleeding. Postoperative mortality was 9.8%. There was no statistically significant difference in either overall morbidity, pancreatic fistula rates, septic complications, bleeding or postoperative mortality between jaundiced and nonjaundiced patients. Preoperative bilirubinemia in patients without PBD and without postoperative complications did not significantly differ from those without PBD who developed complications. Overall morbidity, pancreatic fistula rate, septic complications, bleeding, postoperative mortality were similar in PBD and nonPBD groups as well as in stented and nonstented patients. CONCLUSIONS: As obstructive jaundice has no impact on postoperative results of PD. PBD or stenting of bile ducts should not be used routinely and is justified in cases when surgery with intent for cure has to be postponed. PMID- 12738906 TI - [The assessment of chemotherapy and radiation therapy effect in malignant lymphomas by computed tomography and magnetic resonance imaging]. AB - Aim of this study was to estimate the diagnostic possibilities of computed tomography (CT) and magnetic resonance imaging (MRI) in the assessment of treatment effect in malignant lymphomas. In the period of 1998 to 2002, 196 patients with histologically proven malignant lymphoma were examined in the Department of Tomography of Kaunas University of Medicine Hospital. The data were processed with SPSS 10.1, including application of standardized t-test and classification of Fisher's statistics. In this group the density of lymphomatous masses prior and post treatment has no difference on CT images, thus the differentiation of active tissue and relapse is not possible. On MR images the difference of signal intensity of the active component (not treated) and fibrous remnant is significant. MRI is a radiological method which provides information about activity of lymphomatous tissue and is able to delineate the active component (recidive or partial remission) and inactive remnant (total remission) of the tumor mass and thus facilitating the diagnosis of possible relapse of malignant lymphomas for hematology, chemotherapy and radiotherapy specialists. PMID- 12738905 TI - [The comparative role of computed tomography and magnetic resonance imaging in the diagnosis of extracapsular spread of malignant lymphomatous masses invading blood vessels]. AB - Aim of the study was to search for an optimal method an of investigation in diagnosis of extracapsular spread of the malignant lymphomas and invading the blood vessels. In the period of 1998 to 2002, 81 patients with malignant lymphomas with coverage of neck and body areas were examined in the Department of Tomography of Kaunas University of Medicine Hospital. It was performed by computed tomography (CT) and magnetic resonance imaging (MRI), with or without iv. application of contrast media. The data were processed with SPSS 10.1 (Statistical package for Social Sciences 10.1 for Windows), including application of chi(2), t-test. Specificity, sensitivity and diagnostic accuracy of CT and MRI methods were calculated and compared according to recommendations by Gefland D. W. and Ott D. J., 1985. Diagnosis of extracapsular spread of the lymphomatous tissue and invading the blood vessels was best performed by MR method (specificity, sensitivity, accuracy in this case 91-95%). Bolus CT angiography because of low resolution in the range of soft tissues, insufficient opacification of blood vessels with contrast medium and differences in blood flow was not informative enough (specificity, sensitivity, accuracy in this case 80 85%). PMID- 12738907 TI - [Treatment of supracondylar humerus fractures in children, according to Gartland classification]. AB - Supracondylar fractures are the most common fractures in the area of elbow in children. The purpose of this study was to review and analyze the treatment patterns of supracondylar humerus fractures in childhood. There were 93 children with supracondylar humerus fractures treated in the Department of Pediatric Orthopedics within the period from March 2000 till November 2002. Ninety fractures were extension-type injuries, 3 were flexion injuries, these were not included in our study. Supracondylar humerus fractures were classified according to Gartland classification. Type III fractures were found in 63 patients, type II fractures were seen in 23 patients and 4 patients had type I fractures. Indications for treatment were determined according to fracture type. Four children were treated with external immobilization alone. Twenty-three patients underwent closed reduction and percutaneous pinning by K-wires. Sixty-two patients were treated with closed reduction and external immobilization such as a plaster cast or according to Blount. Eleven of these 62 children (type III) underwent close or open reduction and internal fixation in follow-up. In 1 case of comminutive fracture a patient underwent immediate operation. There were no early or late complications. All operated patients healed without sequela and had excellent or good results. We had noticed that all manipulations should be performed immediately in order to avoid severe swelling and neurological or vascular complications. In type I fractures we performed the external immobilization only. Type II displaced fractures can be treated satisfactorily with closed reduction and external immobilization. Type III displaced fractures should be treated with closed reduction and percutaneous pinning with K-wires under the fluoroscope guidance. It is believed to be a safe, reliable and efficient method for treatment of this difficult fracture. Indications for open reduction and internal fixation includes open fractures, fractures complicated by vascular injury, unsatisfactory closed reduction due to unstable fracture. PMID- 12738908 TI - [The treatment and complications of intraarticular fractures of the distal radius]. AB - The purpose of this trial was to determine the rate of complications and the cause of complications after intraarticular fractures of the distal radius. We examined those patients, who had intraarticular fractures of the distal radius and who were treated in Kaunas University of Medicine Hospital and Kaunas Red Cross hospital in 1998-2000. The evaluation of results was made by movement of the joint and pain in the wrist, which was evaluated by pain scale and McGill pain questionnaire. Palmar slope, radial inclination and radial length were evaluated rentgenologically. Distal radioulnar joint was evaluated as well (with evaluation of scapholunate ligament condition). Patients were investigated 1.5 and 2 years after fracture. RESULTS: There are more secondary dislocations and the shortening of radius in group that was treated by gypsum bandage. Good distraction of fracture ends is seen using external fixation, but strain of soft tissue gives complications: contraction of fingers, and instability of wrist. The incongruence of joint surface causes early arthrosis and pain. The injury of ligaments in the wrist, which can be seen by X-ray examination, causes instability of wrist and chronic pain. PMID- 12738909 TI - [Combined coronary artery bypass grafting and carotid endarterectomy: early postoperative results and perspectives]. AB - OBJECTIVE: The aim of this study was to review early postoperative results of the combined coronary artery bypass grafting and carotid endarterectomy (CABG/CEA) performed at the Clinic of Cardiac Surgery Heart Center of Kaunas University of Medicine. MATERIAL AND METHODS: A series of consecutive 19 patients (64.5+/-8.2 years old, 17 (89.5%) male patients) with coexisting severe ischemic cardiac and carotid artery disease were operated on during 8-year period ending December 2001. Seven (36.8%) patients with carotid artery stenosis were asymptomatic. Previous neurological events had occurred in 12 (63.2%) patients: five (26.4%) patients presented with a previous stroke, seven (36.8%) - with a transient ischemic attack. Demographics and perioperative variables of these patients were compared with variables of 42 patients, that underwent isolated CABG during this period (control group). RESULTS: After introducing of the routine preoperative screening carotid ultra-sonography in the year 2000, the rate of asymptomatic carotid artery stenosis among all diagnosed hemodynamically significant carotid stenoses has raised up to 43.8%. The in-hospital results were similar between the two groups with regard to 30-day perioperative mortality, stroke and major postoperative complications. CONCLUSIONS: Based on our results, combined interventions of CEA and CABG can be performed with an acceptable morbidity and mortality when severe carotid stenosis is associated with advanced, symptomatic ischemic heart disease. Patients with total occlusion of the carotid artery compose a high-risk subgroup for perioperative stroke. The introduction of routine preoperative carotid duplex scanning resulted in higher rate of asymptomatic carotid artery disease among the patients scheduled for combined CABG/CEA procedure. PMID- 12738910 TI - [Course of pregnancy, delivery and newborn status in case of maternal tuberculosis]. AB - Course of pregnancy, delivery and newborns status were analyzed for 77 women with tuberculosis. The data were compared with analogical data of 72 healthy women. Almost fifty percent (49.3%) of patients were treated with antituberculous drugs in pregnancy. Eighty percent of women with tuberculosis gave birth of mature healthy newborns. Some cases of preterm rupture of membranes, preterm or postterm delivery and another complications were observed, but there was no significant difference comparing to healthy women. Anemia and congenital anomalies were significantly more common in tuberculosis group. Anemia was explained by suppressive influence on the appetite and liver function of pregnant women by specific baciliar toxicosis, antituberculous drugs and insufficient resorption of nutrition ingredients (especially iron). We are very concerned about growing number of developmental anomalies of newborns in women with tuberculosis, such as spina bifida, inherited luxation of hip joint and other abnormalities. There is a need for additional data to make a general conclusion. But it is clear that in medical institutions all women with tuberculosis must get the information how to plan pregnancy. PMID- 12738911 TI - [Spiroveloergometry parameters and parameters of blood gases and acid-base equilibrium at rest and during exercise in athletes]. AB - The aim of the study was to evaluate spiroveloergometric and blood gases data and compare them among swimmers and basketball players. Examinations were performed for swimmers and basketball players (n=85). Spirometry at rest and spiroveloergometry, analysis of blood gases and acid-base equilibrium at rest and during exercise were performed. Minute ventiliation of swimmers at rest was statistically significantly (p<0.001) lower than of basketball players: 11.0 l/min and 22.1 l/min correspondingly. Partial pressure of blood carbon dioxide for swimmers was larger, than of basketball players (41.5 and 35.9 mm Hg correspondingly, p<0.001), plasma bicarbonates as well (25.2 and 22.5 mmol/l, p<0.01) and there was no bases excess (BE -0.22 and -2.95, p<0.001). Spiroveloergometry showed, that at the same minute ventiliation and the same work level oxygen metabolic units (MET) were significantly (p<0.001) larger for swimmers: 21.2 and 14.9 correspondingly. Decompensated metabolic acidosis during maximal exercise for swimmers was not determined. CONCLUSIONS: swimmers' ventilation was more economic than of basketball players, oxygen consumption and aerobic capacity were statistically significantly better. PMID- 12738912 TI - The cumulation of Wild pansy (Viola tricolor L.) accessions: the possibility of species preservation and usage in medicine. AB - Wild pansy (Viola tricolor L.) has a history in folk medicine of helping respiratory problems such as bronchitis, asthma, and cold symptoms. The drugs and extracts are prepared from raw material of pansy; it is a component of some prepared antitussives, cholagogues, dermatological medicines, roborants and tonics, alternatives, and anti-phlebitis remedies. Wild pansy is indigenous to or naturalized in large parts of Europe and the Middle East as far as Central Asia, also found through the United States. In the Lithuanian flora wild pansy habitats areas have been fast reducing; this not only limits the availability of the reserves of medicinal raw materials for pharmacy and therapy needs but also causes a menace to survival of species. The reasons of reduction of natural habitats and areas of wild pansy are not only unfavorable meteorological conditions (including summer droughts) but also the competition of different herbs and irrational human activities. The opportunities of preservation of the species wild pansy need to be cultivated and the most exhaustive adaptation research should be performed. PMID- 12738913 TI - [Long-term risk of myocardial infarction after a first-ever stroke]. AB - PURPOSE: To examine the incidence of myocardial infarction during the first three years after a first-ever stroke. MATERIAL AND METHODS: Both Kaunas community based stroke register and ischemic heart disease register served at the primary source of data. The methods used for data collection were those applied by the World Healt Organisation (WHO) for the WHO MONICA project. During 1986 to 1994, 2531 patients aged 25 to 64 years with a first-ever stroke were registered and followed-up for three years for the information on the first-ever acute myocardial infarction or death from ischemic heart disease. Actuarial life tables were used to analyze risk of myocardial infarction. RESULTS: During the study period, 58 patients with the first-ever acute myocardial infarction that occurred among the first-ever stroke survivors were identified. Fifty-three first-ever ischemic stroke survivors (39 (73.6%) men and 14 (26.4%) women, (p=0.07)) experienced their first-ever myocardial infarction. The cumulative risk of suffering an acute myocardial infarction was 3.5% (95% confidence interval 2.6 to 4.4%) by 3 years: in male ischemic stroke survivors this risk accounted for 4.3% (95% confidence interval 3.0 to 5.6%) and in female - 2.3% (95% confidence interval 1.1 to 3.5%). The risk was the highest one early after ictus in the ischemic stroke survivors: 0.5% (95% confidence interval 0.2 to 0.8%) by 3 months (men - 0.7% (95% confidence interval 0.2 to 1.2%), women - 0.3% (95% confidence interval 0.0 to 0.7%)). CONCLUSION: The risk of myocardial infarction is the highest early after the first stroke, although due to relatively small number of cases confidence intervals were too wide to reach a statistical significance. PMID- 12738914 TI - [Venous air embolism]. AB - Venous air embolism is the entry of air into the venous system as a consequence of trauma or iatrogenic complications (especially central venous cannulation or pressurized intravenous infusion systems). It also can occur following the surgical procedures. Venous air embolism results in right ventricular dysfunction and pulmonary injury. In this review article various causes, frequency, pathophysiology, clinical features, diagnosis, treatment, outcome and prevention of venous air embolism are discussed. PMID- 12738916 TI - [The use of masking level differences in evaluating central auditory processing: the norms for the normal hearing subjects]. AB - OBJECTIVES: This study sought to determine whether scores of masking level differences (MLD) were influenced by age and sex and to derive norm values from normal hearing subjects for the Turkish population. STUDY DESIGN: A total of 100 normal hearing subjects were tested in two age groups. Each group consisted of 25 females and 25 males. The mean ages were 21 (range 17 to 24 years) and 31 years (range 25 to 40 years) in group 1 and group 2, respectively. Following pure-tone threshold and speech discrimination tests, the MLD test was performed by using 500 Hz pure-tone and narrow-band noise centered around 500 Hz. The results were analyzed by the ANOVA test. RESULTS: The mean MLD score was 10.92 dB. The upper and lower limits of MLD scores between two standard deviations were 6 and 14 dB, respectively. No significant differences were found between MLD scores with regard to age and sex. CONCLUSION: Since MLD scores are not affected by age and sex, the norm values obtained can be utilized in the investigation of some pathologic conditions. PMID- 12738915 TI - [The effect of vitamin E on histopathologic healing and lipid peroxidation levels in experimentally induced traumatic tympanic membrane perforations]. AB - OBJECTIVES: To examine the favorable effects of vitamin E on tympanic membrane perforations induced mechanically in guinea pigs. STUDY DESIGN: Bilateral tympanic membrane perforations of 1.8 mm were induced in 40 guinea pigs. The animals were randomly divided into two groups equal in number. One group remained untreated, while the other was administered vitamin E (100 mg/kg/day) through intramuscular injections. On days 1, 3, 5, and 7, five animals in each group were randomly sacrificed. Histopathologic changes in the tympanic membranes were evaluated and malondialdehyde levels were determined. RESULTS: Significant increases were observed in epithelial thickness, fibroblastic proliferation, and neovascularization in the study group (p<0.05). Epithelial thickness was found to be increased in both groups beginning from the first day; however, this increase was more rapid in the study group. Although malondialdehyde levels showed significant increases on days 3 and 5 in both groups (p<0.05), they returned to the first day values in vitamin E-treated animals on day 7, whereas controls still maintained high malondialdehyde levels. CONCLUSION: Vitamin E hastens the healing process of traumatic tympanic membrane perforations. PMID- 12738917 TI - [A comparison between findings of high resolution computed tomography and magnetic resonance imaging in the preoperative assessment of cochlear implant patients]. AB - OBJECTIVES: We compared the preoperative findings of high resolution computed tomography (HRCT) and magnetic resonance imaging (MRI) with those observed during surgery to determine their value for cochlear implant patients. PATIENTS AND METHODS: A retrospective assessment was made on preoperative HRCT and MRI scans of 124 cochlear implant patients (71 males, 53 females; mean age 15 years; range 2 to 63 years). Congenital cochlear anomalies, cochlear ossification, new bone formation in the middle ear, and structures in the internal auditory canal were evaluated and compared with intraoperative findings. RESULTS: Cochlear anomalies detected in five patients by HRCT and MRI were confirmed intraoperatively. Cochlear ossification encountered in six patients during surgery was only demonstrated in four patients; HRCT and MRI scans only showed a narrow basal turn in the remaining two patients. During surgery, eight patients were found to have cochlear fibrosis: this finding was documented in five patients (62.5%) by MRI, and in none by HRCT scans. CONCLUSION: Although HRCT provides valuable information for surgical planning, its limitations may justify the additional use of MRI before cochlear implantation. PMID- 12738918 TI - [Comparison of adenoid and tonsil core cultures in chronic adenotonsillitis]. AB - OBJECTIVES: Tonsil and adenoid core cultures were compared and beta-lactamase producing bacteria were determined in patients with chronic adenotonsillitis. PATIENTS AND METHODS: Thirty-two patients (21 boys, 11 girls; mean age 5 years) with chronic adenotonsillitis underwent elective adenotonsillectomy. The core swaps of tonsil and adenoid tissues were obtained under sterile conditions and were inoculated in 5% sheep blood agar, eosin methylene blue agar, and chocolate agar plates. The frequency of beta-lactamase producing isolates were assessed. RESULTS: Staphylococcus aureus was the most common pathogen both in tonsil and adenoid core cultures. Among pathogenic bacteria isolated from tonsil (n=27) and adeonid (n=22) cultures, the same strains were isolated in 20 cultures (75%). The frequencies of beta-lactamase producing bacteria were 44% and 41% in tonsil and adenoid core cultures, respectively. S. aureus was associated with beta-lactamase production in 88% and 100% in tonsil and adenoid tissues, respectively. CONCLUSION: Differences between tonsil surface and core bacterial flora may have implications in the etiopathogenesis and treatment of chronic tonsillitis. The range of species isolated seems to be similar for both chronic tonsillitis and adenoiditis. PMID- 12738919 TI - Spontaneous cerebrospinal fluid rhinorrhea associated with empty sella: a transnasal-transsphenoidal repair of the fistula. AB - Spontaneous cerebrospinal rhinorrhea is a rare clinical condition. More commonly it develops due to cerebral tumors or hydrocephalus; it is seldom seen in association with empty sella syndrome. A sixty-three-year-old woman who presented with left nasal discharge was hospitalized on the suspicion of CSF rhinorrhea. Analysis of the nasal fluid revealed a clear color, a positive Pandy's test, and a positive result for glucose (72%). On magnetic resonance scans, herniation of the suprasellar cistern was observed into the sellar cavity, and the left sphenoid sinus was filled with contrast medium. The repair of the fistula was performed via a transnasal and transsphenoidal approach. The patient was followed up for five years without any symptoms or recurrences. PMID- 12738920 TI - [Rhinophyma treated by excision and full-thickness skin grafting under local analgesia]. AB - A seventy-three-year-old man presented with a complaint of enlarged nose. The diagnosis was made as rhinophyma and the mass was excised under local anesthesia. The involved tissues including the columellar, alar, and dorsal nasal skin were removed by preserving the perichondrium and periosteum. A full-thickness skin graft was obtained from the supraclavicular region and transferred to the defect area with a tie-over dressing. An acceptable texture and color adaptation of the skin graft was obtained at the end of one-year follow-up period. PMID- 12738921 TI - [Ethmoid sinus osteoma with orbital extension]. AB - Osteoma is the most common neoplasm of the paranasal sinuses. A fifty-year-old woman who had presented with complaints of protrusion of the eyeball and double vision was referred to our department with a diagnosis of proptosis. Computed tomography of the paranasal sinuses and the orbits showed a high-density mass of 2x3 cm, involving the left ethmoidal cells laterally, the left maxillary sinus superomedially, the left orbit inferomedially, and the nasal cavity superiorly. The mass was removed through the external ethmoidectomy approach. Postoperatively, all visual functions improved. The pathologic diagnosis was osteoma. PMID- 12738922 TI - [A review on experimental sinusitis models in rabbits]. AB - Experimental animal models are instrumental to study the pathogenesis of diseases and to examine the results of treatment. The great size and ease of accessibility have made rabbit sinuses popular in the investigation of experimental sinusitis. Microbiologic, histopathologic, and radiologic aspects of this model have been well-described and the effects of medical and surgical treatments on sinusitis have been examined. Ongoing research may provide further data to find more simple and non-invasive methods to induce sinusitis in rabbits that may more closely simulate sinusitis pathogenesis in humans. This article reviews the most recent data on technical, surgical, microbiologic, histopathologic, and radiologic aspects, and complications of sinusitis models in rabbits. PMID- 12738923 TI - [Day Surgery. An opportunity for postgraduate surgeons' training]. AB - BACKGROUND: It is widely acknowledged that day surgical procedures must be performed by expert personnel or carried out under their supervision. Recent regulations introduced by Schools of Postgraduate Surgery oblige postgraduate surgeons to carry out a number of minor and medium-sized operations. The characteristics of day surgery make it an appropriate organisational model for complying with the requirements of these directives. Interventional research has shown that adequate training permits education to be coupled with good results. METHODS: The postgraduate surgeons are trained to perform hernioplasty using Trabucco's method through surgical training based on an initial phase of at least 20 operated patients during which they act as the second surgeon, and a phase of 10 patients during which they are the first surgeon working alongside a tutor with proven experience. Five doctors are completing their training, having acted as first surgeon assisted by a tutor during the period 01/11/1999 - 31/10/00 in 60 hernioplasty operations using Trabucco's method out of a total of 143 carried out in the same period, with an average of 12 operations each. Specific informed consent was obtained from all patients. All operations were performed under local anesthesia. RESULTS: There was no need to convert to general anesthesia. Complications were comparable to those occurring during surgery performed by experienced colleagues. No recurrences were reported. The mean follow-up was 10 months. CONCLUSIONS: An evaluation of these data confirms the real possibility of dedicating a number of ambulatory surgical procedures to training postgraduate surgeons. This would not lead to a reduced efficacy of treatment and does not increase the percentage of complications. PMID- 12738924 TI - [Day Surgery. Five years of experience and activity]. AB - BACKGROUND: The aim of this paper is to evaluate the management of an integrated unit of Day Surgery in a General Surgery Department. METHODS: An outline of the first 5 years of activity is presented. Setting of investigation: little provincial hospital. Patients have been selected on a basis of Day Surgery specific features: a total of 1,294 patients. Pathologies treated: inguinal hernias, phlebectasias, phimosis, borsitis, arthritic cysts, tendon cysts, carpal tunnel, Dupuytren's disease ("crispatura tendinum"), lymphadenitis, mammary nodules, hemorrhoids, hydroceles, varicoceles, adipomas. Patients' selection parameters, surgical operation type and modalities, postoperative course, protected discharges from hospital, follow-up and complications have all been carefully recorded. RESULTS: The most numerous operations were related to hernial pathology (54.32%) and to adipomas (10.81%). The form of anesthesia was mainly loco-regional. Only 4 cases (0.3%) had to be hospitalized for the night after operation. COMPLICATIONS: 1 serious wound infection, needing removal of the hernial prosthesis; 1 painful "tumefaction" on the inguinal wound for hernioplastic operation; 3 "seromas" in inguinal hernioplastics. CONCLUSIONS: Advantages of Day-Surgery: cut in health costs due to the reduction of admissions to hospital; reduction in hospital infections and in surgical wounds; reduction in drug use; thinning of waiting lists; increase of available beds for other pathologies; reduction of disability days; high appreciation index. PMID- 12738925 TI - [Gastric stromal tumors. Our experience in 21 cases surgically managed]. AB - BACKGROUND: A retrospective review on 21 patients with gastric stromal tumors who underwent surgical treatment in the period 1974-2001, is presented. METHODS: The patients were 8 males and 13 females, with an average age of 62.6 years. Histological examination showed 15 smooth muscle tumors (9 benign and 6 malignant), 5 neural tumours (3 benign and 2 malignant) and 1 undifferentiated tumor. Main symptoms were abdominal pain and palpable abdominal mass and the most sensible diagnostic techniques were endoscopy and abdominal CT-scan. Surgical procedures performed were: local resection (15 cases), partial gastric resection (3 cases), subtotal gastrectomy (2 cases) and total gastrectomy (1 case). RESULTS: There were no operative mortality and morbidity. Among 12 patients with benign GSTs, 1 died for causes not related to the disease, while 11 patients are still alive and in good health after a mean follow-up of 148.5 months (range 6 262). Among patients with malignant muscular GSTs, 3 were lost to follow-up and 3 are alive and free from disease after a median follow-up of 58 months (range 3 108). The 2 patients with malignant neural GSTs are still alive and in good health 140 and 24 months after surgical procedure, while the patient with undifferentiated GST died for liver metastases 43 months after total gastrectomy. CONCLUSIONS: The authors conclude that the most frequent symptoms are abdominal pain and a palpable mass, but no specific signs have been detected. Endoscopy plays a very important diagnostic role and CT-scan is the most sensible technique in the evaluation of location, size, invasion of adjacent organs and metastasis. The aim of treatment must be the complete resection of the tumor and the prognostic prediction on the basis of histologic findings is quite difficult. PMID- 12738926 TI - [Anastomotic leakage after traditional surgery of the colon and rectum]. AB - BACKGROUND: Anastomotic leakage after traditional surgery of the colon and rectum represents one of the more frightening complications for its high incidence (5 10%) leading to a mortality of 10-33%. The aim of the study is to analyze the main etiopathogenetic factors of this complication in the literature and, retrospectively, in personal experience. METHODS: At the General Surgery Department of the Hospital of Imperia, between September 1997 and August 2001, 278 laparotomic procedures with large bowel anastomosis were completed; 37 patients (13.3%) were treated in emergency. There were 153 males (55%) and 125 females (45%), the average age being 70.5 years (range 41-94). The mean hospital stay was 12 days (range 7-45). RESULTS: The incidence of perioperative mortality was 1.1% (3 cases). Anastomotic leakage occurred in 4 patients (1.4%); one of these died. Analysis of the presented series confirms that factors contributing to leakage of colonic anastomoses are: distal rectum anastomoses, rectal cancer, experience of the surgeon and preoperative radiotherapy. Anastomotic leakage didn't occur in emergency, probably for a prudent attitude that considers multiple-stages surgical procedures in high-risk patients; besides a radical subtotal or total colectomy was performed in the treatment of obstructive cancer of the left colon and rectum. CONCLUSIONS: The incidence of anastomotic leakage in colo-rectal surgery was lower when anastomoses were performed in ideal circumstances; it is necessary to know the main risk factors to reduce this complication. PMID- 12738927 TI - [Endoscopic palliative treatment of the common bile duct at the hepatic hilum. Results in 583 patients treated in a single center over a 10-year period]. AB - BACKGROUND: The outcomes of endoscopic biliary drainage for malignant stenoses at the hepatic hilum were retrospectively evaluated. METHODS: From January 1990 to June 2001, 583 patients, 368 males, average age 69+/-18.5 years, were recruited. Endoscopic procedure consisted of insertion of 1 ore multiple stents, plastic or metallic, across the stricture, under mild sedation. RESULTS: Successful stent insertion was achieved in 518/583 (88.8%) patients and successful drainage in 474 (81.3%) patients. Early complications were observed in 101 (17.3%) patients with related-mortality of 17 (2.9%) patients. Late complications occurred in 39.9% of patients. Survival was of 189 days, on average. CONCLUSIONS: Endoscopic palliation should be the initial management of choice for malignant biliary stenoses at the hepatic hilum. PMID- 12738928 TI - [Surgical treatment of pilonidal sinus with open surgical technique]. AB - BACKGROUND: Personal experience and results about the surgical treatment of pilonidal sinus using the open technique operation are reported. METHODS: Between January 1984 and June 2001, 577 patients (457 males and 120 females) underwent this surgical technique. Eighty-five patients presented recurrences of the under lying disease previously treated with closed technique in other centres. In order to reduce the risks of recurrence, pilonidal sinus was widely resected in order to remove every fistulous track laterally or towards the anal edge. The surgical wound has to be wide, carefully drained, without vertical walls: the outer part of the wound should be wider than its bottom. The persistence of a hairless scar is a suitable result to avoid relapses. RESULTS: This surgical technique allowed very good RESULTS: Postsurgical morbidity was not observed in these patients; the median time to achieve healing was 8+/-2 weeks. Furthermore, after a 3 year follow-up, only 4 patients (0.69%) relapsed. Recurrent dressings are needed; anyway they neither represent a limit of this technique nor lead to a long term absence from working. The postsurgical period of resting is quite similar to that usually prescribed after other surgical techniques and mainly depends on the patient nature. The treatment with connective tissue repairing drugs may be useful to obtain an effective healing. CONCLUSIONS: In the authors' experience, the open surgical technique may obtain the definitive removal of pilonidal sinus, even if particularly wide, with a safe postsurgical course and a very hight percentage (99.31%) of complete healing without any relapse. PMID- 12738929 TI - [Diabetes and bleeding in thoracic surgery]. AB - BACKGROUND: The aim of this study is to assess the influence of diabetes over early postoperative bleeding in thoracic surgical patients. In fact, diabetes leads to hypercoagulation as well as to an alteration of microvessels that could have a negative effect on the retraction and vasoconstriction of the damaged microvessel before hemostasis coagulation phase. METHODS: Data referring to 193 typical pneumonectomies associated with extensive removal of mediastinic nodes, 19 performed in diabetic patients have been retrospectively analysed. RESULTS: Any statistically significant difference between the two groups was found. CONCLUSIONS: More studies would be necessary to confirm our conclusions, on more extensive series of patients with more severe diabetic disease, as well as on non thoracic surgical patients, in order to avoid the consequences of the early and sudden negative pressure on wounds, that in thoracic patients could hide the effects of less evident factors. PMID- 12738930 TI - Colorectal tumor screening. From occult blood testing to genetic evaluation. AB - In Italy 250,000 subjects every year are affected by malignant tumors, causing 150,000 deaths/year. The global index (statistic prevalence) of oncological patients, either at first diagnosis, or restored to health or under treatment, aggregates 1 million and 500 thousand individuals. During his whole lifetime, every male can be exposed to the risk of tumor developing, up to 1/3 times against 1/5 of a female. At the same time it is true that just only in a decade, 5-year survival expectance for this kind of patients has significantly grown: in Italy, during the late '70s only 33% of them reached this survival, becoming 39% in the late '80s. This aim was achieved also through the recruitment of the sensibilization to the prevention and the early diagnosis, properly known as "screenings", which, from the original experiences regarding gynecology, (female breast and uterus), are involving more and more anatomic districts. The purpose of this brief is illustrating the procedures, either usually validated ones, or affirming ones, to identify colorectal neoplasms at early stady or precancerous intestinal lesions. PMID- 12738931 TI - [The role of surgery in the treatment of hepatic metastases from colorectal cancer]. AB - Colorectal neoplasms are a common and frequently fatal illness. Presence of distance metastases from colorectal neoplasm does not preclude therapeutic treatments. Surgical resection is the standard treatment for hepatic colorectal metastasis. Some good results in hepatic metastasectomy are due to the progress of radiology which allows not only early findings of metastatic pathology but defines a surgical technique planning. Patients who are candidates for surgical resection are those with no extrahepatic pathology, resectable hepatic metastases with 1 cm of disease free margin and adequate residual parenchyma. Recurren-ces are shown in two thirds of surgical patients; this suggests that microscopic pathology persists commonly even after resection and that adjuvant therapy is critical. In recent years, many palliative techniques for hepatic cancers have been developed. Potentially useful role of these techniques is ablation of small lesions in patients with contraindications to hepatic resection, small recurrences not resectable, and not resectable neoplasms diffuse in both lobes. Accurate follow-up is essential after hepatic metastasectomy. In conclusion, patients with hepatic potentially resectable colorectal metastases should be evaluated by an expert surgeon, because better long-term outcome is derived from surgical resection. PMID- 12738932 TI - [Unusual localizations of hydatid cyst. Epidemiological aspects and diagnostic problems. Description of a clinical case]. AB - A rare case of primary hydatid cyst of the mesentery is described. Though cystic echinococcosis infestation can occur in any part of the body primary peritoneal involvement is rare. The mechanism of primitive peritoneal infection by the parasite is still unclear. The hypotheses which can explain this rare primary localization, the sensitivity and the specificity of the serological and instrumental tests and the therapeutic management of this atypical lesion are discussed. PMID- 12738933 TI - [Four cases of mesenteric cystic neoformations]. AB - Four cases of mesenteric cystic neoformations personally observed in the last years are reported. This pathology should not be underestimated, since not only does it present several problems regarding diagnosis and treatment, but a nosologic classification is also difficult to make. Since such cysts present with aspecific symptoms, a definite diagnosis cannot always be reached by a preliminary X-ray investigation, which may lead to various interpretations, but often requires a subsequent histologic examination of operative specimen. PMID- 12738934 TI - [Adenocarcinoma of pancreas with situs viscerum inversus totalis]. AB - A case of adenocarcinoma of the head of the pancreas in a patient with situs viscerum inversus totalis, an association described for the third time in literature, is reported. The possible coexistence of malformations of transposed organs and the specular anatomosurgical situation requires particular attention in the diagnosis and preoperative evaluation as well as a careful reorientation of the surgical perspective and a correct surgical conduct. PMID- 12738935 TI - Management of true splenic artery aneurysms. Two case reports and review of the literature. AB - Few series of splenic artery aneurysms (SAA) have been reported, but today asymptomatic SAA are detected with increasing frequency. Their importance lies from their potentially fatal consequences as life-threatening hemorrhage. SAA management still remains controversial as reported in this review. Our 2 patients treated with resection of the aneurysms, both located in the middle third of the splenic artery. Some authors demonstrated that when splenic artery has been ligated (or embolized) and the patients remain anatomically splenic, they may not retain any splenic function. Laparoscopic SAA ligation repair appears to be optimal and useful for aneurysms protruding from the pancreas and it is gaining interest because clinical recovery is rapid with a poor morbidity and economic and cosmetic advantages. Transcatheter embolization too offers a temporary control in urgency to stop hemorrhage and go back at later date to make much better elective operation. Endovascular interventions as percutaneous embolization has recently gained popularity: it offers a safe alternative or adjunctive therapy to traditional surgery. We hope in the future instrumentation will likely improve so that this procedure can be done percutaneously by development of prosthetic devices in the 21th century. PMID- 12738936 TI - Supra infection of amoebic liver abscess consequent to acute appendicitis. Clinical case. AB - Amoebic liver abscess is the most common extra-intestinal manifestation of amebiasis with approximately 10% of the world's population infected by this parasite. Actually, incidence of this infection is also increasing in industrialized countries, as a consequence of the more frequent immigration or travelling. Only 3-10% of patients with intestinal amebiasis develop liver abscess. A clinical case of suprainfection of amoebic liver abscess consequent on acute appendicitis is presented. PMID- 12738937 TI - [Giant ovarian cyst. A case weighing 23 kg (50.6 lb). Literature review]. AB - The ovarian cysts are one of the most common affections for females. Besides non neoplastic functional cysts (follicular or luteal) with relatively small sizes, the neoplastic types, generally benign and that can be ascribed to serous tumors, of 10-15 cm or even bigger should be included. Nowdays, these neoformations are diagnosed relatively early, before they become of big dimensions, even if often they present few symptoms. Then it seems to be interesting to report a case recently observed of an ovarian cyst of 35x45x50 cm and weighing 23 kg in 29-year old woman. PMID- 12738938 TI - Cerebral palsy diagnosis and management: the state of the art. PMID- 12738940 TI - Awakenings. PMID- 12738941 TI - The psychological impact of maxillofacial trauma: an overview of reactions to trauma. AB - We provide an overview of the current understanding of posttraumatic psychological problems, such as posttraumatic stress disorder, and the implications for maxillofacial surgeons and allied professionals. Posttraumatic stress disorder is not confined to combat veterans; it is common after all traumatic events-including maxillofacial injury-and can become chronic unless recognized and treated. The neurobiologic underpinnings of and the known vulnerability factors for posttraumatic stress disorder are increasingly understood, assisting the nursing and surgical staff in their assessments. Both psychological and pharmacologic treatments have been shown to have efficacy; nevertheless, intervention by mental health professionals will not be required by all. PMID- 12738942 TI - Fibrin sealants and platelet preparations in bone and periodontal healing. AB - Fibrin sealants and platelet concentrates have been used alone or in association with bone substitutes to promote bone healing in orthopedic and oral surgery. Commercial fibrin sealants are homologous plasma-derived products that mimic the last step of a coagulation cascade, leading to a fibrin clot. They are used for topical hemostasis and tissue sealing and as melting agents for particulate bone substitutes. Infectious risk led to the development of autologous fibrin sealants from the patient's own plasma. However, their fabrication results in less reproducible or less satisfactory rheologic properties. The use of autologous products with high platelet concentrations, such as platelet-rich plasma, platelet concentrate, and platelet gels, has recently been suggested for combining the fibrin sealant properties with the growth factor effects of platelets. We describe the characteristics and limitations of fibrin sealants (commercial/autologous) and platelet preparations and review their effects on bone and periodontal healing as reported in experimental studies and clinical trials. PMID- 12738943 TI - Asymptomatic submandibular mass. PMID- 12738944 TI - Patient satisfaction after trigeminal nerve repair. AB - OBJECTIVE: The purpose of this study was to measure patient satisfaction and to evaluate the factors influencing patients' perceptions of the outcome of inferior alveolar nerve or lingual nerve repair. STUDY DESIGN: We used a retrospective cohort study design and a sample of patients who underwent repair of inferior alveolar nerve or lingual nerve injuries. The major outcome variable was the patient's overall satisfaction with treatment. The patient's satisfaction was rated as either good to excellent (group A) or fair to poor (group B). RESULTS: The study sample was composed of 46 patients with a mean age of 28 +/- 12 years; 76% were female. Fifty-five percent of the sample reported their overall satisfaction to be good to excellent. No individual predictor factors were statistically associated with patient satisfaction. Among the outcome variables, the measures of taste, pronunciation, self-consciousness, and function were statistically significantly different (P <.05) between the 2 groups. CONCLUSIONS: After nerve repair, more than half of the patients rated their overall satisfaction with the operative results to be good to excellent. PMID- 12738945 TI - Visually guided irrigation for patients with symptomatic internal derangement of the temporomandibular joint: a preliminary report. AB - OBJECTIVE: Visually guided irrigation (VGIR) is simple joint irrigation with minimally invasive intracapsular inspection through the use of a thin arthroscope. The aims of the present study were to develop VGIR for the superior joint compartment of the temporomandibular joint (TMJ) by using a 1.2-mm-diameter rod-lens arthroscope and to examine the short-term clinical outcomes. The correlation between the clinical outcome of VGIR of the TMJ and intracapsular findings was also evaluated. STUDY DESIGN: Twenty patients who underwent VGIR for symptomatic internal derangement of the TMJ (n = 20) were enrolled in this study. The clinical outcome of VGIR was evaluated in terms of the improvement in the painless range of mandibular motion (ROM: interincisal distance in millimeters), and functional jaw pain was evaluated by using a visual analog scale (VAS) ranging from 0 to 100. Painless ROM and VAS scores 6 months postoperatively were compared with the preoperative data. A good clinical outcome was defined as an increased ROM (ROM > 38 mm) and a decreased VAS score (VAS score < 20%, and <60% of the preoperative level). With respect to these criteria, all joints were classified into either good clinical outcome (GO) or poor clinical outcome (PO) groups. The difference in the distribution of the arthroscopic findings obtained during VGIR, with respect to severity, was analyzed between the GO and PO groups. RESULTS: None of the 20 patients experienced any serious local or systemic complications, including the breakage of instruments. We were able to perform a reliable intracapsular diagnosis during VGIR. Sixteen of 20 patients (80%) had significant improvements in postoperative painless ROM and the VAS score of functional pain, thereby being classified into the GO group. The distribution of the severity of intracapsular findings did not differ significantly between the GO group and the PO group. CONCLUSIONS: VGIR is useful for the treatment and diagnosis of internally deranged TMJs. These data suggest that the clinical outcome of TMJ irrigation is not related to the intracapsular condition of the superior joint compartment. PMID- 12738946 TI - Visually guided temporomandibular joint irrigation in patients with chronic closed lock: clinical outcome and its relationship to intra-articular morphologic changes. AB - OBJECTIVE: The aim of this study was to evaluate the clinical outcome after visually guided irrigation (VGIR) of the temporomandibular joint (TMJ) and its relationship with postoperative arthroscopic changes. STUDY DESIGN: Of the original 69 patients, thirty patients (30 TMJs) underwent VGIR of the TMJ a second time. After the first VGIR, the clinical outcome was assessed, and 18 patients were assigned to the good outcome group. The remaining 12 patients were assigned to the poor outcome group. The arthroscopic findings related to the articular surface, synovial lining, and fibrous adhesion scores were recorded. Then, the arthroscopic findings in the first and second VGIR were compared. RESULTS: The intra-articular tissue status between the first and second VGIR was unchanged in approximately 40% of all joints. No significant differences with respect to an improvement in tissue status were found when the good outcome and poor outcome groups were compared. CONCLUSIONS: In patients with chronic closed lock of the TMJ, a clinical improvement after VGIR does not seem to be accompanied by improved intra-articular tissue status. PMID- 12738947 TI - Meta-analysis of periodontal disease and risk of coronary heart disease and stroke. AB - OBJECTIVE: The purpose of this study was to analyze published studies and abstracts in order to provide a quantitative summary of periodontal disease as a risk factor for cardiovascular disease and to explore the possible causes for conflicting results in the literature. STUDY DESIGN: We searched all published literature on the Medline literature search engine since 1980. An additional search was performed with bibliographic citations from each article. Nine cohort studies (8 prospective and 1 retrospective), in which relative risks (RRs), CIs, and P values were reported or could be calculated were included. Four researchers independently extracted RRs, CIs, and P values from each study and evaluated the degree of confounding adjustment. The combined result was calculated with weighted average, and sources of disparity were tested with regression analyses. RESULTS: The summary RR was 1.19 (95% CI, 1.08 1.32), indicating a higher risk of future cardiovascular events in individuals with periodontal disease compared with those without. In an analysis stratified to individuals of /= 50% reduction in seizure frequency from baseline) and number of seizure- free patients versus placebo. In addition, efficacy appears to be maintained over the long term and no evidence for the development of tolerance to the effects of levetiracetam has been seen. (c) 2001 Prous Science. All rights reserved. PMID- 12738961 TI - Syntaxin specificity of cytokinesis in Arabidopsis. AB - Syntaxins interact with other SNAREs (soluble NSF-attachment protein receptors) to form structurally related complexes that mediate membrane fusion in diverse intracellular trafficking pathways. The original SNARE hypothesis postulated that each type of transport vesicle has its own distinct vesicle-SNARE that pairs up with a unique target-SNARE, or syntaxin, on the target membrane. However, recent evidence suggests that small G-proteins of the Rab family and their effectors mediate the initial contact between donor and acceptor membranes, providing complementary specificity to SNARE pairing at a later step towards membrane fusion. To assess the role of syntaxin specificity in membrane recognition requires a biological assay in which one syntaxin is replaced by other family members that do not normally function in that trafficking pathway. Here, we examine whether membrane fusion in Arabidopsis thaliana cytokinesis, which involves a plant-specific syntaxin, the cell-cycle-regulated KNOLLE (KN) protein, can be mediated by other syntaxins if expressed under the control of KN cis regulatory sequences. Only a non-essential syntaxin was targeted to the plane of cell division and sufficiently related to KN to perform its function, thus revealing syntaxin specificity of cytokinesis. PMID- 12738965 TI - Future trends in the treatment of cognitive disorders. AB - This review provides an overview of the different types of cognitive disorders, with special emphasis on the description on Alzheimer's disease, as statistical data indicate that it is the major cause of dementia or cognitive decline. The factors responsible for biochemical changes at the cellular level and the different therapies available for the treatment of cognitive disorders are described. Recent advances, including the elucidation of biochemical changes at the molecular level and different approaches to treat the disease, are also briefly reviewed. (c) 2001 Prous Science. All rights reserved. PMID- 12738966 TI - Short dementia questionnaire for assessing the severity of cognitive impairment in patients with dementia. AB - This paper describes the Short Dementia Questionnaire (SDQ), a psychometric test that provides an initial assessment of the severity of cognitive impairment in patients with dementia. This test requires minimal effort on the part of the assessor and places little strain on the patient. For several years now, a preliminary form of the test has been used for large-scale screening of dementia patients in nursing homes within clinical drug trials. This test is short, inexpensive and largely standardized in regard to implementation, even without the usual intensive training. This paper focuses in particular on the structure and implementation of the test. The item and scale parameters relevant for the evaluation of its field of application are presented, and finally, the results of an initial validation study are reported. These results show that the test procedure is a good predictor of severity scores in frequently used, more extensive tests for grading the severity of dementia. (c) 2001 Prous Science. All rights reserved. PMID- 12738967 TI - The influence of acquired intelligence on cerebral impairment in the elderly. AB - During a psychopharmacological trial, the inpatients of three geriatric nursing homes underwent a psychometric examination in which the Multiple Choice Intelligence Test (MWT-B) was used to determine the level of premorbid intelligence. The results seem to indicate that older people with mild to moderate organic brain syndrome and healthy elderly individuals show little difference with regard to their cognitive plasticity. Shaping conditions of the social sphere, as well as a performance-stimulating environment which continuously encourages independent behavior, appear to be of decisive importance for the protection of specific cognitive abilities and mobilization of reserve capacities. (c) 2001 Prous Science. All rights reserved. PMID- 12738968 TI - Osteoporosis. PMID- 12738969 TI - Lipid lowering therapy as an important adjunct to stroke prevention in coronary heart disease patients. AB - Lipid altering pharmacotherapy has been shown to reduce stroke in coronary heart disease (CHD) patients. Several mechanisms of cerebrovascular protection attributed to these agents include reduction in embolic stroke from cardiac, aortic and carotid sites, delayed progression of carotid stenoses, stabilization of vulnerable carotid atherosclerotic plaque and improvement in cerebral blood flow. (c) 2001 Prous Science. All rights reserved. PMID- 12738970 TI - Targets to trials in diabetic complications. AB - Diabetes is now the leading cause of end-stage renal disease, blindness, lower extremity amputations and impotence. In addition, the risk of death from cardiovascular disease such as myocardial infarction and stroke is more than doubled in diabetics. In September 2001, scientists from around the world met in Melbourne to discuss the mechanisms underlying neuronal and vascular changes in diabetic complications. This report summarizes the meeting and attempts to identify potential targets for drug intervention in diabetic complications. (c) 2001 Prous Science. All rights reserved. PMID- 12738971 TI - Tenecteplase (TNK tissue plasminogen activator): A new fibrinolytic for the acute treatment of myocardial infarction. AB - Tenecteplase, a new fibrinolytic with longer plasma half-life and greater specificity for fibrin than alteplase, achieves rapid thrombolysis in patients with acute myocardial infarction. Similar TIMI grade 3 flow rates at 90 minutes have been observed with tenecteplase and alteplase. The efficacy and safety of tenecteplase have been confirmed in a large trial, ASSENT-2, demonstrating similar mortality rates at 30 days and a lower rate of noncerebral bleeding complications when compared with alteplase. The convenience of a single bolus treatment may facilitate the initiation of early and efficient reperfusion. (c) 2001 Prous Science. All rights reserved. PMID- 12738972 TI - Irritable bowel syndrome: A drug development minefield. PMID- 12738973 TI - Esomeprazole: A significant advance beyond omeprazole in the treatment of acid related disease. AB - The second-generation proton pump inhibitor (PPI) esomeprazole is a new chemical entity consisting of an optical isomer of omeprazole, which for many years has been acknowledged as the gold standard therapy in gastric acid-related disorders. Esomeprazole has demonstrated a unique pharmacokinetic profile and enhanced efficacy over omeprazole, with improved inter-patient pharmacokinetic consistency and a similar safety profile. Esomeprazole has been tested as a therapeutic agent in the management of erosive esophagitis, symptomatic gastroesophageal reflux disease (GERD) and, in combination with appropriate antibiotic therapy, for the eradication of the Helicobacter pylori bacterium and healing of H. pylori associated duodenal ulcers. In clinical studies, esomeprazole has shown greater efficacy than omeprazole with a comparable low incidence of adverse events. (c) 2001 Prous Science. All rights reserved. PMID- 12738974 TI - Faslodex(TM) for the treatment of breast cancer. AB - Faslodex(TM) (fulvestrant), also known as ICI 182780, is the first in a new class of selective estrogen receptor down-regulators (SERDs), which target and degrade the estrogen receptor (ER), and has been developed for the treatment of advanced breast cancer. Up to now application of tamoxifen, a partial estrogen antagonist, has been the "gold standard" in breast cancer therapy in postmenopausal women. However, breast tumors become resistant to tamoxifen after a while, leading to progression of the cancer. Also, the risk of the development of endometrial carcinoma is one of the disadvantages in treatment with tamoxifen. Therefore "pure" antiestrogens with high affinity to the estrogen receptor, but without agonistic activity, have been developed in the last few years. Summarizing all data from in vitro and in vivo studies and clinical trials, the antiestrogen Faslodex(TM) (AstraZeneca, Cheshire, U.K.) appears to be a very promising new agent for the treatment of advanced and early breast cancer. (c) 2001 Prous Science. All rights reserved. PMID- 12738975 TI - Recent research in gastroesophageal reflux disease. PMID- 12738976 TI - Hypertensive disorders of pregnancy: Canadian renaissance. PMID- 12738977 TI - Imaging the endometrium: are there predictors of uterine receptivity? AB - Ultrasound imaging technology brings new insight to cyclic changes in the endometrium and offers the potential to assess the probability of embryo implantation in natural and assisted reproduction cycles. However, the data reported are diverse and frequently conflict in their analysis and conclusions. This review examines imaging techniques used for endometrial evaluation, including grey-scale ultrasonography and colour-flow, power-flow, and spectral Doppler interrogation of the uterine vasculature. New techniques being used for uterine assessment, such as computer-assisted image analysis, three-dimensional ultrasonography, and magnetic resonance imaging, are also discussed. Each of these techniques is under active investigation and has the potential to provide rapid, noninvasive endometrial evaluation. Currently, there is reasonable certainty that women with heterogeneous endometrial linings of less than 6 mm will rarely conceive, and that endometrial contractions, or lack thereof, play some role in the successful establishment of pregnancy, even though this role has not yet been identified. Given the set of new imaging technologies available, it is feasible to expect that visual assessments will someday be used as predictors of uterine receptivity. PMID- 12738978 TI - Serious perinatal complications of non-proteinuric hypertension: an international, multicentre, retrospective cohort study. AB - OBJECTIVE: To determine the proportion of births complicated by either a pre existing or a gestational non-proteinuric hypertension, presenting at <34 weeks' gestation, and the associated incidence with 1 or more serious perinatal complications or birth weight <3rd centile for gestational age. METHODS: A retrospective chart review was conducted in 5 international centres, from 1998 to 2002, where "tight" control (normalization) of blood pressure (BP) is the norm. International Classification of Diseases (ICD) codes were used to identify women who delivered at > or =20 weeks' gestation, with any hypertensive disorder of pregnancy. Women were included if they had a diastolic blood pressure (dBP) of 90 to 109 mm Hg, due to either a pre-existing or a gestational non-proteinuric hypertension, presenting at <34 weeks' gestation. Women were excluded if they had ongoing severe hypertension, or if at presentation with dBP of 90 to 109 mm Hg, they had 1 or more of the following: proteinuria, an indication for "tight" control of BP or imminent delivery, or a known intrauterine fetal death or lethal fetal anomaly. Data were collected on paper forms, scanned into an electronic database, and summarized descriptively by type of hypertension. RESULTS: There were 305 eligible women (0.7% deliveries, 12.8% hypertensive deliveries) identified with non-proteinuric hypertension that was either pre-existing (133 [43.6%]) or gestational (172 [56.4%]). Regardless of hypertension type, 16.4% (n = 50) of pregnancies were complicated by birth weight <3rd centile or 1 or more serious perinatal complications, 34.3% (n = 100) by preterm birth, 30.8% (n = 94) by preeclampsia, and 2.0% (n = 6) by serious maternal complications. CONCLUSION: Non-proteinuric pre-existing or gestational hypertension, presenting before 34 weeks' gestation, identifies a subpopulation of hypertensive pregnant women at both substantial perinatal risk and maternal risk. The CHIPS (Control of Hypertension In Pregnancy Study) trial is designed to determine how best to manage the hypertension of such women in order to optimize perinatal outcome. PMID- 12738979 TI - Eclampsia in Southern Alberta: is there a role for seizure prophylaxis in all women with gestational hypertension? AB - OBJECTIVE: To evaluate the predictability of eclampsia and explore the role for seizure prophylaxis in a population with a low frequency of seizure prophylaxis. METHODS: A retrospective review was conducted of all women with eclampsia registered at the Foothills Hospital in Calgary, Alberta, between 1991 and 2000. The data collected included timing of seizure in relation to diagnosis of gestational hypertension (GHTN) and delivery, method of seizure prophylaxis (if any), and maternal characteristics. RESULTS: During the study period, 3075 of 38,577 women (8.0%) were diagnosed with GHTN, with or without proteinuria or adverse conditions. Three percent had received magnesium sulfate for seizure prophylaxis. Of these 3075 women, 17 (0.6%) developed eclampsia, none of whom was receiving magnesium sulfate for seizure prophylaxis at the time. Of these, 10 women (59%) exhibited GHTN prior to their first seizure, including 6 women with GHTN with adverse conditions, 3 with GHTN with proteinuria but without adverse conditions, and 1 with GHTN without proteinuria or adverse conditions. Five of the 17 women had seizures that occurred prior to labour, 6 were intrapartum, and 6 were postpartum. Nine (53%) of the 17 women with eclampsia had their initial seizure after the diagnosis of GHTN and before 24 hours postpartum. CONCLUSION: Seizure prophylaxis for all the women with GHTN, from the time of diagnosis through 24 hours postpartum, may have been able to prevent as many as 53% of eclamptic episodes. Three hundred and seven women with GTHN would have to receive seizure prophylaxis to prevent one seizure. PMID- 12738980 TI - Exposure to emergency contraception in an undergraduate medical curriculum. AB - OBJECTIVES: To determine senior medical students' exposure to lectures and clinical experiences on emergency contraception (EC). METHODS: Ninety third-year medical students at the University of Western Ontario (UWO) were surveyed at the end of a year of clinical clerkship. The survey assessed exposure to contraception, EC, and other reproductive health experiences over the preceding 3 years of medical training, as well as personal opinions about the availability of family planning services. Survey data were analyzed using Statistical Package for the Social Sciences (SPSS 10.0) software. RESULTS: Seventy-three medical students (47% female) completed the questionnaire, giving a response rate of 81.1%. Classroom teaching of EC was reported by 71.2% of the cohort. Only 16.9% had counselled women about EC themselves; 33.8% had not been exposed to it at all in a clinical setting. A majority of students (92.5%) stated they would provide services for contraception, but only 7.5% felt such issues were well addressed in the undergraduate medical program. CONCLUSION: Undergraduate medical trainees at UWO perceive that the subject of contraception in general and EC in particular are not well addressed. PMID- 12738982 TI - Mechanisms of action of phenanthroperylenequinones in photodynamic therapy (review). AB - Despite the age-old belief that most anti-cancer agents kill tumor cells by necrosis, recent findings have demonstrated that photosensitizers could also kill tumor cells by triggering genetically programmed series of events termed apoptosis. Cell death by apoptosis is a very neat way to eliminate unwanted cells: no traces are left and the cell contents are never released or accessible to the immune system. Hence there is no inflammation. This is in contrast to death by necrosis. Under these conditions, normally the cell swells and then, when membrane integrity comes under attack, the cell collapses like a balloon and the contents spill out into the extracellular milieu. This may result in an inflammatory response. Because of the relatively clean nature of the apoptotic process, it is desirable to identify compounds that effectively activate the apoptotic pathway. Photodynamic therapy (PDT), a new mode of treatment, is based on the combined use of light-absorbing compounds and light irradiation. Recent developments in understanding the mechanisms of the PDT effect of photosensitizers indicate that a critical factor in the success of the agent is the ability to induce apoptosis in the malignant cell population. Hypericin and Hypocrellins are perylquinones, which are novel natural photosensitizers characterized by high absorption around 470 nm and high singlet oxygen yield. To study the signaling mechanism in vitro we have investigated uptake kinetics, intracellular localization, mode of cell death and mechanisms involved in the photodynamic action following PDT in human cell lines of poorly differentiated (CNE2) and moderately differentiated (TW0-1) nasopharyngeal carcinoma (NPC) and also poorly differentiated colon (CCL-220.1) and bladder (SD) cells. PMID- 12738981 TI - The management of uterine leiomyomas. AB - OBJECTIVE: The objective of this document is to serve as a guideline to the investigation and management of uterine leiomyomas. OPTIONS: The areas of clinical practice considered in formulating this guideline are assessment, medical treatments, conservative treatments of myolysis, selective artery occlusion, and surgical alternatives including myomectomy and hysterectomy. The risk-to-benefit ratio must be examined individually by the woman and her health care provider. OUTCOMES: Implementation of this guideline should optimize the decision-making process of women and their health-care providers in proceeding with further investigation or therapy for uterine leiomyomas, having considered the disease process and available treatment options, and reviewed the risks and anticipated benefits. EVIDENCE: English-language articles from MEDLINE, PubMed, and the Cochrane Database were reviewed from 1992 to 2002, using the key words "leiomyoma," "fibroid," "uterine artery embolization," "uterine artery occlusion," "uterine leiomyosarcoma," and "myomectomy." The level of evidence has been determined using the criteria described by the Canadian Task Force on the Periodic Health Examination. BENEFITS, HARMS, AND COSTS: The majority of fibroids are asymptomatic and will not require intervention or further investigations. For the symptomatic fibroid, hysterectomy offers a definitive solution. However, it is not the preferred solution for women who wish to preserve their uterus. The predicted benefits of alternative therapies must be carefully weighed against the possible risks of these therapies. In the properly selected woman with symptomatic fibroids, the result from the selected treatment should be an improvement in the quality of life. The cost of the therapy to the health-care system and to women with fibroids must be interpreted in the context of the cost of untreated disease conditions and the cost of ongoing or repeat treatment modalities. RECOMMENDATIONS: 1. Medical management should be tailored to the needs of the woman presenting with uterine fibroids and geared to alleviating the symptoms. Cost and side effects of medical therapies may limit their long-term use. (III-C) 2. In women who do not wish to preserve fertility and who have been counselled regarding the alternatives and risks, hysterectomy may be offered as the definitive treatment for symptomatic uterine fibroids and is associated with a high level of satisfaction. (II-A) 3. Myomectomy is an option for women who wish to preserve their uterus, but women should be counselled regarding the risk of requiring further intervention. (II-B) 4. Hysteroscopic myomectomy should be considered as first-line conservative surgical therapy for the management of symptomatic intracavitary fibroids. (I-B) 5. It is important to monitor ongoing fluid balance carefully during hysteroscopic removal of fibroids. (I-B) 6. Laparoscopic myolysis may present an alternative to myomectomy or hysterectomy for selected women with symptomatic intramural or subserous fibroids who wish to preserve their uterus but do not desire future fertility. (II-B) 7. Uterine artery occlusion may be offered as an alternative to selected women with symptomatic uterine fibroids who wish to preserve their uterus. (I-C) 8. Women choosing uterine artery occlusion for the treatment of fibroids should be counselled regarding possible risks, and that long-term data regarding efficacy, fecundity, pregnancy outcomes, and patient satisfaction are lacking. (III-C) 9. Removal of fibroids that distort the uterine cavity may be indicated in infertile women, where no other factors have been identified, and in women about to undergo in vitro fertilization treatment. (III-C) 10. Concern of possible complications related to fibroids in pregnancy is not an indication for myomectomy, except in women who have experienced a previous pregnancy with complications related to these fibroids. (III-C) 11. Women who have fibroids detected in pregnancy may require additional fetal surveillance when the placenta is implanted over or in close proximity to hen the placenta is implanted over or in close proximity to a fibroid. (III-C) 12. In women who present with acute hemorrhage related to uterine fibroids, conservative management consisting of estrogens, hysteroscopy, or dilatation and curettage may be considered, but hysterectomy may become necessary in some cases. (III-C) 13. Hormone replacement therapy may cause myoma growth in postmenopausal women, but it does not appear to cause clinical symptoms. Postmenopausal bleeding and pain in women with fibroids should be investigated in the same way as in women without fibroids. (II-B) 14. There is currently no evidence to substantiate performing a hysterectomy for an asymptomatic leiomyoma for the sole purpose of alleviating the concern that it may be malignant. (III-C) VALIDATION: This guideline was reviewed and accepted by the Clinical Practice Gynaecology Committee, and by Executive and Council of the Society of Obstetricians and Gynaecologists of Canada. SPONSOR: The Society for Obstetricians and Gynaecologists of Canada. PMID- 12738983 TI - RB-resistant Abl kinase induces delayed cell cycle progression and increases susceptibility to apoptosis upon cellular stresses through interaction with p53. AB - c-Abl, a non-receptor tyrosine kinase, is found in both nucleus and cytoplasm of proliferating fibroblasts. RB negatively regulates the kinase activity of c-Abl. Overexpression of kinase active c-Abl can overcome RB-induced growth arrest in Saos-2 cells. However, we previously reported that disruption of the RB matchmaker function leads to delayed cell cycle progression in the presence of p53. In this study, we investigated whether overexpression of mutant c-Abl (AS2, RB-resistant Abl kinase) not only lead to delayed cell cycle progression but also make cells susceptible to apoptosis under coexpression with a fragment of RB C pocket in human skin fibroblast. AS2 expressing cells showed delayed cell growth rate in normal growth condition. After genotoxic stress such as etoposide treatment, AS2 expressing cells readily progressed into apoptosis through p53 and caspase-3 activations. Our results suggest that expression of AS2 not only induces delayed cell cycle progression but also results in increased sensitivity to apoptosis in the presence of p53. PMID- 12738984 TI - Over-expression of DNA methyltransferases and CDKN2A gene methylation status in squamous cell carcinoma of the oral cavity. AB - The present study was designed to investigate the potential relationship between CDKN2A (p16) gene hypermethylation, which has reported to be frequently observed in oral squamous cell carcinomas (OSCCs), and expression of human DNA methyltransferases (DNMTs: DNMT1, DNMT3A and DNMT3B). Twenty-five pairs of primary OSCCs and matched normal oral mucosa tissues were examined. The p16 gene was hypermethylated (48%) in the tumors showing significant down-regulation of both mRNA and protein expressions. A demethylation assay on 8 OSCC-derived cell lines was also performed by means of treatment with the demethylating agent, 5 aza-2'-deoxycytidine. Four of 5 cell lines showing down-regulation of the p16 gene, revealed re-activation of gene expression after the treatment. In contrast, frequent over-expression of DNMT mRNA expression, also found in the expression of the proteins, was detected: DNMT1 at 72% and DNMT3A at 56%, and DNMT3B at 64%, respectively. However, we could not identify any statistical significance between p16-hypermethylation status in individual tumors and the expression of any of the three DNMTs. These data suggest that hypermethylation of the p16 gene and up regulation of DNMTs are involved in oral carcinogenesis, but they may be through different mechanisms. PMID- 12738985 TI - Early diagnosis of malignant melanoma: Proposal of a working formulation for the management of cutaneous pigmented lesions from the Melanoma Cooperative Group. AB - Epiluminescence microscopy (ELM) strongly improves the separation of different types of cutaneous pigmented lesions (CPL) and facilitates the early diagnosis of cutaneous melanoma (CM). ELM alone is not 100% accurate in routine diagnosis, and should not be considered the only criterion in the diagnosis of high-risk skin lesions. We have however, demonstrated close agreement between ELM classification criteria and histology in 2,731 cutaneous lesions. In the past five years, our Melanoma Cooperative Group has evaluated 61,000 skin lesions from 30,000 individuals and identified 478 cutaneous melanomas. Most newly diagnosed patients had very early stage melanoma [299 (62%) were Stage I (203 Stage IA and 96 Stage IB), by the American Joint Committee on Cancer (AJCC) criteria]. We have compared data from the patient histories and clinical evaluations with ELM-based morphological patterns to better characterize skin lesions and minimize interpretative problems. From these comparisons, we propose new guidelines for the management of CPL to provide a standard diagnostic and therapeutic approaches and to foster the early identification of lesions at risk for malignant transformation. PMID- 12738986 TI - Identification of a novel human gene, ZFP91, involved in acute myelogenous leukemia. AB - To elucidate mechanisms leading to acute myelogenous leukemia (AML), to find sensitive markers and novel targets for drug therapy, and to allow choice of suitable chemotherapy for each affected individual, we previously compared expression of mRNA from mononuclear cells of AML patients with that of normal controls using a cDNA microarray. Data from that study identified many genes that were commonly up- or down-regulated in AML cells. Of these, we report here the identification of a novel gene whose expression was increased in 27 (93%) of the 29 AML cases whose PBMC preparations include >70% leukemia cells. The gene product, localized in nuclei, showed several characteristics of transcription factors: five zinc-finger domains, a leucine zipper, and several nuclear localization signals. Its 92.5% identity in amino-acid sequence to the murine penta zinc finger protein (mPZf; gene symbol Zfp91), led us to term it ZFP91. Anti-sense oligonucleotides inhibited expression of ZFP91, suppressed cell growth, and induced apoptosis. Our results suggest that ZFP91 is likely to play an important role in cell proliferation and/or anti-apoptosis, and may serve as a molecular marker for AML. PMID- 12738987 TI - Heterogeneous gene alterations in primary breast cancer contribute to discordance between primary and asynchronous metastatic/recurrent sites: HER2 gene amplification and p53 mutation. AB - The aim of the present study was to clarify differences in genetic events between primary breast cancers and asynchronous metastatic/recurrent lesions, by examining HER2 gene amplification and p53 mutation. The subjects were 44 breast cancer patients with asynchronous metastasis or recurrence. Synchronous metastases were excluded. HER2 overexpression and gene amplification were examined using immunohistochemistry and fluorescent in situ hybridization (FISH). P53 point mutation was examined by immunohistochemistry, laser-captured microdissection, PCR-single-strand conformation polymorphism, and a direct sequencing method. Immunohistochemistry showed that, for HER2, p53, ER and PgR, discordance rates between primary and recurrent tumor were 2 (4.5%), 1 (2.3%), 7 (15.9%) and 10 (22.7%), respectively. Two primary tumors with discordant HER2 overexpression were composed of at least two populations of carcinoma cells, with and without HER2 gene amplification. Distribution of HER2 gene amplification was consistent with protein overexpression. Corresponding recurrent tumors consisted of carcinoma cells without HER2 gene amplification. Of 6 recurrent tumors in which the primary carcinoma had a p53 point mutation, 3 tumors had identical mutations, 1 tumor had a different point mutation, and 2 tumors had no mutation. It was suspected that the latter 3 recurrent tumors comprised a minor component of the primary tumor. In the present study, we examined a large series of asynchronous recurrent tumors. A limited number of these tumors showed discordance between primary and recurrent tumors. Detailed observations revealed that cell populations present in recurrent tumors were also present in the primary tumors, although they comprised a minor component of the primary tumor. Heterogeneity of the primary tumor apparently contributed to discordance. PMID- 12738988 TI - In vivo detection of resistance to anthracycline based neoadjuvant chemotherapy in locally advanced and inflammatory breast cancer with technetium-99m sestamibi scintimammography. AB - Neoadjuvant chemotherapy (NACT) is the treatment of choice in patients (pts) with locally advanced (LABC) and inflammatory breast cancer (IBC). To evaluate the role of Tc-99m sestamibi imaging in the prediction of response to NACT and in the in vivo functional detection of intrinsic or acquired chemoresistance, 24 female pts with LABC (n=21) or IBC (n=3) were prospectively studied. Tc-99m scintimammography was performed 1-3 days before treatment (first) and 2-5 days after the completion (second) of NACT (epirubicin and cyclophosphamide for LABC and doxorubicin and vinorelbine for IBC). Three planar images (2 lateral prone and one anterior supine, 10 min/each) were obtained 10 min postinjection and a lateral prone image (10 min) of the affected breast (B) was obtained at 4 h. To calculate the tumor to normal B ratio (TBR), 2 identical irregular regions of interest (ROIs) were drawn around the tumor (T) and in adjacent ipsilateral normal B on both early (E) and delayed (D) prone lateral images. The TBR was obtained as the ratio of the mean counts per pixels in the 2 ROIs. Then Tc-99m sestamibi retention index (RI) in the T was determined by dividing the D-TBR by the E-TBR. Afterwards, NACT response was assessed pathologically or clinically in inoperable disease. Scintigraphic sensitivity for correct prediction of T presence after NACT was 81% (17/21), whereas specificity for correct prediction of T absence was 100% (3/3). In LABC, 3 patients had a pathological complete response: first RI was high (>0.56) in all 3, while no T uptake was visible on the second scintigraphy. Eighteen patients did not show a pathological complete response: in 5, both first and second RI were low (60 years (p<0.05), high grade histology (p<0.001), presence of B symptoms (p<0.005), nodal presentation (p<0.005) and advanced stage at diagnosis (p<0.001). Histological sub-type did not significantly correlate to outcome. In conclusion whilst a number of prognostic indicators can assist in determining the outcome in PTCL, these lymphomas are complex and often follow an unpredictable course. In order to make the best clinical decisions in individual cases, more clinical study is required. PMID- 12739007 TI - MGC9753 gene, located within PPP1R1B-STARD3-ERBB2-GRB7 amplicon on human chromosome 17q12, encodes the seven-transmembrane receptor with extracellular six cystein domain. AB - MYC, ERBB2, MET, FGFR2, CCNE1, MYCN, WNT2, CD44, MDM2, NCOA3, IQGAP1 and STK6 loci are amplified in human gastric cancer. It has been reported that the gene corresponding to EST H16094 is co-amplified with ERBB2 gene in human gastric cancer. Here, we identified and characterized the gene corresponding to EST H16094 by using bioinformatics. BLAST programs revealed that EST H16094 was derived from the uncharacterized MGC9753 gene. Two ORFs were predicted within human MGC9753 mRNA, and ORF1 (nucleotide position 18-980 of NM_033419.1) was predicted as the coding region of human MGC9753 mRNA based on comparative genomics. Nucleotide sequence of mouse Mgc9753 mRNA was next determined in silico by modification of AK052486 cDNA (deleting C at the nucleotide position 37). Human MGC9753 and mouse Mgc9753 proteins were 320-amino-acid seven-transmembrane receptors with the N-terminal six-cysteine domain and an N-glycosylation site (85.0% total-amino-acid identity). Human MGC9753 protein showed 90.6% total-amino acid identity with human CAB2 aberrant protein, which lacked the third transmembrane domain of MGC9753 due to frame shifts within ORF. Human MGC9753 gene, consisting of eight exons, were clustered with PPP1R1B, STARD3, TCAP, PNMT, ERBB2, MGC14832 and GRB7 genes within the 120-kb region. PPP1R1B, STARD3, MGC9753, ERBB2 and GRB7 genes are co-amplified in several cases of gastric cancer. This is the first report on comprehensive characterization of the amplicon around the PPP1R1B-STARD3-TCAP-PNMT-MGC9753-ERBB2-MGC14832-GRB7 locus on human chromosome 17q12. PMID- 12739008 TI - FLJ10261 gene, located within the CCND1-EMS1 locus on human chromosome 11q13, encodes the eight-transmembrane protein homologous to C12orf3, C11orf25 and FLJ34272 gene products. AB - The CCND1-EMS1 locus on human chromosome 11q13 is amplified in esophageal cancer, bladder tumors, and breast cancer. During analyses of FGF gene cluster within the CCND1-EMS1 locus, we identified a 5'-truncated partial cDNA (NM_018043.1) derived from the uncharacterized FLJ10261 gene. Here, we characterized the FLJ10261 gene by using bioinformatics. NM_018043.1 cDNA corresponded to the nucleotide position 1129-4258 of 4558-bp DKFZp686O1156 cDNA, and the nucleotide position 50-3010 of DKFZ-p686O1156 was the coding region of the FLJ10261 gene. FLJ10261 gene, consisting of 26 exons, was located between FGF3 and FADD genes within the CCND1 EMS1 locus. Two FLJ10261 isoforms with or without exon 15 were transcribed due to alternative splicing. FLJ10261 mRNA was expressed in head and neck tumors, parathyroid tumors, breast, pancreatic, and gastric cancer. Mouse Flj10261 gene (AK052589) was located between Fgf3 and Fadd genes on mouse chromosome 7. Human FLJ10261 gene was homologous to C12orf3 gene on human chromosome 12p13, C11orf25 gene on 11p14, and FLJ34272 gene on 12q23. Human FLJ10261 protein showed 89.8% total-amino-acid identity with mouse Flj10261 protein, and also 58.4%, 38.3%, and 38.6% identity with human C12orf3, C11orf25, and FLJ34272/BAC03704 proteins, respectively. FLJ10261, C12orf3, C11orf25 and FLJ34272 proteins were eight transmembrane proteins with N- and C-terminal tails facing the cytoplasm, which might function as transporters for unidentified substrates. This is the first report on comprehensive characterization of the FLJ10261 gene located within the CCND1-ORAOV1-FGF19-FGF4-FGF3-FLJ10261-FADD-PPFIA1-EMS1 locus on human chromosome 11q13. PMID- 12739009 TI - An expression study of hormone receptors in spontaneously developed, carcinogen induced and hormone-induced mammary tumors in female Noble rats. AB - The Noble (Nb) rat model has been used in the study of hormonal carcinogenesis of mammary and prostate glands, as this rat strain is susceptible to tumor induction in these glands by hormonal treatments. Recently, we demonstrated that this rat strain can develop spontaneously mammary tumors at high incidence in aged animals and also show high sensitivity to chemical carcinogens (DMBA and MNU) and combined treatments with sex hormones in mammary tumor induction. In the present study, we examined and compared the expression of hormone receptors [including estrogen receptors (ERalpha and ERbeta), androgen receptor (AR), progesterone receptor (PR), prolactin receptor (PRLR)] and prolactin (PRL) by immunohistochemistry, Western blotting and RT-PCR in spontaneous mammary tumors, and mammary tumors induced by sex hormones (T+E2 and T+DES for 8-10 months) and DMBA in Nb rat model. Immunohistochemistry and Western blotting showed that both the spontaneously developed and hormone-induced carcinomas exhibited strong immunoreactivity of ERalpha, ERbeta, AR, PR and PRLR, while the spontaneous fibroadenomas showed weak to moderate immunoreactivity of ERalpha and PRLR, whereas the DMBA-induced carcinomas exhibited weak to moderate immunoreactivity of ERalpha, AR, PR and PRLR, and sporadic weak ERbeta immunoreactivity. RT-PCR analyses showed that mRNA expression pattern of these markers resembled that of proteins. In addition, weak mRNA expression of PRL was detected in spontaneous carcinomas and carcinomas induced by DMBA and hormones, suggesting that PRL could be produced locally within the tumors. The results showed that the expression status of hormone receptors and PRL was different in spontaneous mammary tumors and tumors induced by carcinogen or hormones, suggesting that the extent of involvement of steroid hormones and their receptors in the spontaneous, carcinogen- or hormone-induced mammary carcinogenesis might be different. PMID- 12739010 TI - Hyperthermia improves cellular immune response to human hepatocellular carcinoma subsequent to co-culture with tumor lysate pulsed dendritic cells. AB - Dendritic cells play a major role in cellular immunity. The crucial steps of antigen presentation and processing by DCs may be limiting factors for adoptive cellular immunotherapy. Here, we investigated whether hyperthermia of human hepatocellular carcinoma (HCC) cells induces enhanced cytotoxic cellular immune response. Peripheral blood mononuclear cell (PBMC)-derived DCs were pulsed with tumor cell lysate of the human HCC cell line HepG2, which had been heat shocked prior to incubation for 5 h. Subsequent to TNFalpha-induced maturation DCs were co-cultured with autologous CD4+ and/or CD8+ cells, and T cell mediated cytolysis of HepG2 cells was assessed. We observed enhanced CD4+/8+ cellular cytotoxicity against HepG2 cells subsequent to co-culture with the heat shocked tumor lysate pulsed DCs as compared to pulsing DCs with lysate of non-heat shocked tumor cells. The improved cellular immune response can be related to enhanced expression of HSP 70 and 90 in HepG2 cells upon hyperthermia. PMID- 12739011 TI - Arthroscopically assisted meniscal allograft transplantation with and without combined anterior cruciate ligament reconstruction. AB - The menisci provide a vital role in load transmission across the knee joint as well as contribute to knee stability, particularly in the ACL-deficient knee. Loss of the meniscus, in part or in total, significantly alters joint function and predisposes the articular cartilage to degenerative changes, which has been well documented both clinically and radiographically. This study examined clinical and patient-reported outcomes following meniscal allograft transplantation with and without combined ACL reconstruction in a select group of 31 patients with complaints of pain and/or instability (34 meniscal allografts); 11 underwent isolated meniscal transplantation and 20 meniscal transplantation combined with ACL reconstruction. Bony fixation was performed with bone plugs for medial transplants and using a bone bridge for lateral transplants. All patients completed several knee-specific and general measures of health-related quality of life and underwent a comprehensive physical examination. Flexion weightbearing PA radiographs at latest follow-up were compared to those obtained preoperatively. Mean follow-up was 2.9 years (range 2-5.5 years). The Activities of Daily Living and Sports Activities Scale scores were 86+/-11 and 78+/-16, respectively, and the average Lysholm score was 84+/-14. There were no significant differences in these scores based upon which meniscus (medial or lateral) was transplanted, concurrent ACL reconstruction, or the degree of chondrosis at arthroscopy. SF-36 scores indicated that patients were functioning at a level similar to the age- and sex-matched population. Twenty-two patients stated they were greatly improved, 8 were somewhat improved, 1 was without change. All but one patient reported that knee function and level of activity were normal or nearly normal. The average loss of motion compared to the noninvolved side was 3 degrees for extension and 9 degrees for flexion. All but one patient had a negative or 1+ Lachman's test. The remaining patient had a 2+ Lachman's test. Assessment with the KT-1000 arthrometer revealed a side-to-side difference of 2 mm (range -2 mm to 7 mm). Average hop and vertical jump indices were both 85% of the contralateral extremity. No statistically significant joint space narrowing was observed by radiography over time. Meniscal allograft transplantation with and without combined ACL reconstruction in carefully selected patients with complaints of compartmental joint line pain and/or instability appears able to provide relief of symptoms and restore relatively high levels of function, particularly during activities of daily living. PMID- 12739012 TI - Effect of combining nicotinamide as a PARS-inhibitor with selective iNOS blockade during porcine endotoxemia. AB - OBJECTIVE: To investigate the effects of combined selective inducible nitric oxide synthase (iNOS) inhibition using 1400 W with nicotinamide (NAD) as a PARS inhibitor on hepato-splanchnic hemodynamics, O(2) kinetics, and energy metabolism during hyperdynamic porcine endotoxemia. DESIGN: Prospective, randomized, controlled, interventional experiment. SETTING: Animal research laboratory. SUBJECTS: Seventeen domestic pigs. INTERVENTIONS: After 12 h of continuous i.v. endotoxin (LPS) infusion 17 pigs received either no drug (CON, n=9) or 1400 W, titrated to maintain mean arterial pressure (MAP) at pre-endotoxin level, plus 10 mg.kg.h NAD ( n=8;). Measurements were obtained before, 12 h, 18 h, and 24 h after starting LPS infusion. MEASUREMENTS AND RESULTS: In addition to systemic and pulmonary hemodynamics and gas exchange, we measured hepatic arterial and portal venous blood flow, liver and portal venous drained viscera O(2) exchange, ileal mucosal-arterial PCO(2) gap, and portal as well as hepatic venous lactate/pyruvate ratios. Expired NO and plasma nitrate levels were assessed as a parameter of NO production. Without affecting cardiac output, therapy maintained MAP and blunted the LPS-induced rise in expired NO levels, attenuated the progressive fall in liver lactate clearance, and blunted the impairment of hepato splanchnic redox state. The rise of ileal mucosal-arterial PCO(2) gap was not influenced. CONCLUSIONS: Combining selective iNOS inhibition with NAD as a PARS blocker may prevent circulatory failure and attenuate the detrimental consequences of LPS in intestinal and hepatocellular energy metabolism. Given the potential hepatotoxicity of high-dose NAD treatment, more potent PARS blockers with higher selectivity might further enhance the benefit of this therapeutic approach. PMID- 12739013 TI - Coronary artery disease in HIV infected patients. AB - OBJECTIVE: To assess the incidence, clinical features, treatment, and follow-up of coronary events in HIV-infected patients over a period of 5 years. PATIENTS AND PARTICIPANTS: A cohort of 840 patients. MEASUREMENTS AND RESULTS: A coronary event occurred in 17 patients (5.9/1000 persons-years). Sixteen of them were exposed to highly active antiretroviral therapy (HAART). Patients with coronary events differed in age (48.3 vs. 43 years), CD4 T-cell count (284 vs. 486/mm(3)), total cholesterol (6.2 vs. 5.3 mmol/l), HDL cholesterol (0.72 vs. 1.16 mmol/l), and LDL cholesterol (4.95 vs. 3.391.61 mmol/l). No difference was observed regarding duration of HAART, weight, glucose level, or smoking status between the two groups. Acute coronary syndrome was the first manifestation in 14 patients. Coronary angiography showed 2.56 stenosis per patient, with a single vessel involvement in one-half. Percutaneous angioplasty was performed in all cases, with stenting in 11. After a mean follow-up of 36 months, 14 patients remain alive. Restenosis ( n=4) occurred in 3 patients (PTCA 3; stenting 4). All 14 patients are free of heart failure symptoms. Their mean left ventricular ejection fraction is 61%. CONCLUSIONS: A higher coronary-event rate is observed among HIV infected patients, associated with drug-induced metabolic disturbances and a high prevalence of tobacco smoking. However, treatment and prognosis of acute myocardial infarction has no specificity. PMID- 12739014 TI - ICU management of severe acute respiratory syndrome. AB - BACKGROUND: Severe acute respiratory syndrome (SARS) is a contagious viral illness first recognized in late 2002. It has now been documented in 26 countries worldwide, with significant outbreaks in China, Hong Kong, Singapore, and Toronto. Research into identifying the etiological agent, evaluating modes of disease transmission, and treatment options is currently ongoing. DISCUSSION: The disease can produce a severe bilateral pneumonia, with progressive hypoxemia. Up to 20% of patients require mechanical ventilatory support, with a fatal outcome occurring in about 5% of cases. CONCLUSIONS: We review the current knowledge about this disease, with particular emphasis on ICU management and infection control precautions to prevent disease transmission. PMID- 12739016 TI - Corneal confocal microscopy: a non-invasive surrogate of nerve fibre damage and repair in diabetic patients. AB - AIMS/HYPOTHESIS: The accurate detection, characterization and quantification of human diabetic neuropathy are important to define at risk patients, anticipate deterioration, and assess new therapies. Corneal confocal microscopy is a reiterative, rapid, non-invasive in vivo clinical examination technique capable of imaging corneal nerve fibres. The aim of this study was to define the ability of this technique to quantify the extent of degeneration and regeneration of corneal nerve fibres in diabetic patients with increasing neuropathic severity. METHODS: We scanned the cornea and collected images of Bowman's layer (containing a rich nerve plexus) from 18 diabetic patients and 18 age-matched control subjects. RESULTS: Corneal nerve fibre density (F(3)=9.6, p<0.0001), length (F(3)=23.8, p<0.0001), and branch density (F(3)=13.9, p<0.0001) were reduced in diabetic patients compared with control subjects, with a tendency for greater reduction in these measures with increasing severity of neuropathy. CONCLUSION/INTERPRETATION: Corneal confocal microscopy is a rapid, non-invasive in vivo clinical examination technique which accurately defines the extent of corneal nerve damage and repair and acts as a surrogate measure of somatic neuropathy in diabetic patients. It could represent an advance to define the severity of neuropathy and expedite assessment of therapeutic efficacy in clinical trials of human diabetic neuropathy. PMID- 12739015 TI - Mutations in the glucokinase regulatory protein gene in 2p23 in obese French caucasians. AB - AIMS/HYPOTHESIS: Glucokinase regulatory protein (GKRP) controls the activity of glucokinase in liver but possibly also in some areas of the central nervous system, suggesting that it could play a role in body mass control. Its gene is located in a region (2p21-23) linked to serum leptin levels. Our goal was to investigate whether mutations in the GKRP gene were associated with obesity. METHODS: Mutations were sought in the GKRP gene of 57 patients from the families of the French genome-wide scan for obesity that contributed most to the positive LOD score with 2p21-23. The identified mutations were further sought in 720 unrelated obese individuals and 384 individuals of normal weight and their effect on the properties of recombinant GKRP were investigated. RESULTS: The most frequent mutation (Pro446Leu) had a similar allele frequency in the obese (0.63) and normal weight (0.64) subjects and did not affect the properties of GKRP. Similarly, no effect on the properties of GKRP was observed with Arg590Tyr, found in 10 out of 720 obese subjects and in 2 out of 384 control subjects (p=0.18). Mutation Arg227Stop was found in one obese family and in 1 out of 384 control subjects and led to an insoluble protein. Mutation Arg518Gln, replacing a conserved residue, led to a marked decrease in the affinity of GKRP for both fructose 6-phosphate and fructose 1-phosphate and to a destabilization of GKRP. However, this mutation did not co-segregate with obesity in the single family in which it was found. CONCLUSIONS/INTERPRETATION: Mutations that affect the properties of GKRP are found in the French population, but they do not seem to account for the linkage between the 2p23 locus and quantitative markers of obesity. PMID- 12739017 TI - Obesity, and not insulin resistance, is the major determinant of serum inflammatory cardiovascular risk markers in pre-menopausal women. AB - AIMS/HYPOTHESIS: Increased serum inflammatory markers have been found in obesity and insulin-resistant states, and could play a causative role in insulin resistance, atherosclerosis and cardiovascular disease. The polycystic ovary syndrome represents a human model of insulin resistance because both lean and obese polycystic ovary syndrome patients are insulin-resistant compared with non hyperandrogenic women. We evaluated whether obesity, insulin resistance, or both, are related to the increased concentrations of inflammatory markers in pre menopausal women. METHODS: We compared 35 patients with polycystic ovary syndrome and 28 healthy women, paired for BMI, prevalence of obesity and smoking. Measurements included serum inflammatory markers, BMI, waist-to-hip ratio, blood pressure, serum glucose, insulin, lipid and hormone concentrations, and insulin sensitivity index. RESULTS: The insulin sensitivity index was reduced in polycystic ovary syndrome patients compared with controls. However, no differences were observed between both groups in C-reactive protein, interleukin 6, tumour necrosis factor-alpha, soluble type 2 tumour necrosis factor receptor, and soluble intercellular cell adhesion molecule-1. When considering patients and controls as a whole, C-reactive protein and interleukin 6, were increased in obese subjects compared with lean women. Inverse correlations existed between insulin sensitivity index and C-reactive protein, interleukin 6, tumour necrosis factor-alpha, soluble type 2 tumour necrosis factor receptor, and soluble intercellular cell adhesion molecule-1. Only the weak correlation with C-reactive protein persisted after controlling for BMI. CONCLUSION/INTERPRETATION: Obesity, and not insulin resistance, is the major determinant of serum inflammatory cardiovascular risk markers in pre-menopausal women. PMID- 12739018 TI - No interactions between polymorphisms in the beta3-adrenergic receptor gene and the PPAR-gamma gene on the risk of the insulin resistance syndrome in the Danish MONICA cohort. PMID- 12739019 TI - Renal disease in relatives of Indo-Asian Type 2 diabetic patients with end-stage diabetic nephropathy. AB - AIMS/HYPOTHESIS: Indo-Asian immigrants in The Hague, The Netherlands, have a nearly 40-fold higher risk of end-stage diabetic nephropathy compared to the Caucasian population. To detect a genetic susceptibility for nephropathy within the Indo-Asian population, we assessed whether familial clustering of nephropathy occurs in families of Indo-Asian Type 2 diabetic patients. METHODS: We compared nephropathy prevalence between two groups of first-degree relatives of Indo-Asian patients with Type 2 diabetes; the first group (case relatives) consisted of 169 relatives of patients with end-stage diabetic nephropathy; the second group (control relatives) consisted of 161 relatives of diabetic patients who had no nephropathy. The case and control relatives were examined for diabetes, blood pressure, renal function, microalbuminuria and urine dipstick measurements. RESULTS: The mean age was 41 years and similar in the case and control relatives. Diabetes was distributed equally in both family groups. We did not find more nephropathy in first-degree relatives of Indo-Asian Type 2 diabetic patients with end-stage diabetic nephropathy in comparison with the control-relatives. CONCLUSION/INTERPRETATION: We could not detect a genetic susceptibility for diabetic nephropathy within the Indo-Asian population. The lack of familial clustering of renal disease in Indo-Asian diabetic patients points to a general genetic or environmental susceptibility for diabetic nephropathy in this population. PMID- 12739020 TI - Ghrelin Arg51Gln mutation is a risk factor for Type 2 diabetes and hypertension in a random sample of middle-aged subjects. AB - AIMS/HYPOTHESIS: Experimental studies have suggested that ghrelin, a novel gastrointestinal peptide hormone, could play a role in glucose homeostasis. In addition, ghrelin has been associated with beneficial haemodynamic effects in experimental settings. Since the Arg51Gln mutation changes the carboxyterminal amino acid of the mature hormone and is associated with low ghrelin concentrations, we assessed the hypothesis that Arg51Gln mutation is a risk factor for Type 2 diabetes, impaired glucose tolerance, and hypertension. METHODS: Blood pressure recordings and oral glucose tolerance test were carried out in the hypertensive ( n=519) and control cohorts ( n=526) of our well-defined OPERA study. The genotypes and plasma IGF-I and IGFBP-1 concentrations of 1031 subjects were analysed. RESULTS: The ghrelin 51Gln allele was a risk factor for Type 2 diabetes, and the effect remained significant after adjustment for age, BMI, and study group (OR=2.53, CI: 1.11-5.75, p=0.027). In addition, the 51Gln allele was a risk factor for hypertension (OR=2.63, CI: 1.37-5.08, p=0.003). 51Gln carriers had lower concentrations of IGF-I and higher concentrations of IGFBP-1 compared to non-carriers. CONCLUSION/INTERPRETATION: The ghrelin 51Gln allele could increase the risk for Type 2 diabetes and hypertension. The low IGF I concentrations in 51Gln carriers suggest that the mechanism might be associated with low GH concentrations. PMID- 12739021 TI - Demonstration of increased concentrations of circulating glycated insulin in human Type 2 diabetes using a novel and specific radioimmunoassay. AB - AIMS/HYPOTHESIS: Glycation of insulin, resulting in impaired bioactivity, has been shown within pancreatic beta cells. We have used a novel and specific radioimmunoassay to detect glycated insulin in plasma of Type 2 diabetic subjects. METHODS: Blood samples were collected from 102 Type 2 diabetic patients in three main categories: those with good glycaemic control with a HbA(1c) less than 7%, moderate glycaemic control (HbA(1c) 7-9%) and poor glycaemic control (HBA(1c) greater than 9%). We used 75 age- and sex-matched non-diabetic subjects as controls. Samples were analysed for HbA(1c), glucose and plasma concentrations of glycated insulin and insulin. RESULTS: Glycated insulin was readily detected in control and Type 2 diabetic subjects. The mean circulating concentration of glycated insulin in control subjects was 12.6+/-0.9 pmol/l ( n=75). Glycated insulin in the good, moderate and poorly controlled diabetic groups was increased 2.4-fold ( p<0.001, n=44), 2.2-fold ( p<0.001, n=41) and 1.1-fold ( n=17) corresponding to 29.8+/-5.4, 27.3+/-5.7 and 13.5+/-2.9 pmol/l, respectively. CONCLUSION/INTERPRETATION: Glycated insulin circulates at noticeably increased concentrations in Type 2 diabetic subjects. PMID- 12739022 TI - Destruction of conditional insulinoma cell lines in NOD mice: a role for autoimmunity. AB - AIMS/HYPOTHESIS: betaTC-tet (H2(k)) is a conditional insulinoma cell line derived from transgenic mice expressing a tetracycline-regulated oncogene. Transgenic expression of several proteins implicated in the apoptotic pathways increase the resistance of betaTC-tet cells in vitro. We tested in vivo the sensitivity of the cells to rejection and the protective effect of genetic alterations in NOD mice. METHODS: betaTC-tet cells and genetically engineered lines expressing Bcl-2 (CDM3D), a dominant negative mutant of MyD88 or SOCS-1 were transplanted in diabetic female NOD mice or in male NOD mice with diabetes induced by high-dose streptozotocin. Survival of functional cell grafts in NOD-scid mice was also analyzed after transfer of splenocytes from diabetic NOD mice. Autoreactive T cell hybridomas and splenocytes from diabetic NOD mice were stimulated by betaTC tet cells. RESULTS: betaTC-tet cells and genetically engineered cell lines were all similarly rejected in diabetic NOD mice and in NOD-scid mice after splenocyte transfer. In 3- to 6-week-old male NOD mice treated with high-dose streptozotocin, the cells temporarily survived, in contrast with C57BL/6 mice treated with high-dose streptozotocin (indefinite survival) and untreated 3- to 6 week-old male NOD mice (rejection). The protective effect of high-dose streptozotocin was lost in older male NOD mice. betaTC-tet cells did not stimulate autoreactive T-cell hybridomas, but induced IL-2 secretion by splenocytes from diabetic NOD mice. CONCLUSION/INTERPRETATION: The autoimmune process seems to play an important role in the destruction of betaTC-tet cells in NOD mice. Genetic manipulations intended at increasing the resistance of beta cells were inefficient. Similar approaches should be tested in vivo as well as in vitro. High dose streptozotocin influences immune rejection and should be used with caution. PMID- 12739023 TI - Circulating anti-pericyte autoantibodies are present in Type 2 diabetic patients and are associated with non-proliferative retinopathy. AB - AIMS/HYPOTHESIS: This study aims to determine the prevalence of anti-pericyte autoantibodies in Type 2 diabetes and to characterize these autoantibodies as new markers of disease activity in diabetic retinopathy. METHODS: A total of 299 patients with Type 2 diabetes participated in this study. Retinopathy was assessed by 7-field stereo fundus photography and was graded according to the ETDRS scale. Serum anti-pericyte autoantibodies were detected by immunofluorescence on tissue cultured bovine retinal pericytes. RESULTS: The prevalence of anti-pericyte autoantibodies in Type 2 diabetic patients was 54% and was approximately equal in men and women. The prevalence was approximately 55% with retinopathy at grades from 10 to 53. At grades above 53 the prevalence declined to 23% ( p<0.0001). The highest prevalence by duration of diabetes, 70%, was found at 0 to 5 years and the lowest, 25% at more than 25 years duration ( p<0.0001). CONCLUSION/INTERPRETATION: Anti-pericyte autoantibodies are present at high prevalence in Type 2 diabetes. Their presence during earlier stages of retinopathy could be due to a reaction with antigens expressed by "activated" pericytes. The decline in antibody prevalence in advanced retinopathy could mark pericyte loss and progression to an angiogenic retinal milieu. PMID- 12739025 TI - Norepinephrine release is reduced in cardiac tissue of Type 2 diabetic patients. AB - AIMS/HYPOTHESIS: The aim of this study was to assess whether cardiac catecholamine release is affected in patients with Type 2 diabetes mellitus. METHODS: A trial tissue was obtained from 19 diabetic (Type 2) and 43 non diabetic patients undergoing coronary surgery. Endogenous norepinephrine release was examined under baseline conditions as well as during electrical field stimulation (effective voltage 5 V, stimulation frequency 4 Hz, pulse width 2 msec) by high performance liquid chromatography and electrochemical detection. Cardiac function and arterial blood pressure was assessed from coronary angiography. RESULTS: In atrial tissue from diabetic patients, stimulation induced norepinephrine release was reduced by 25% compared with non-diabetic patients, while baseline norepinephrine release did not differ between both groups. Preoperative plasma glucose and haemoglobin A(1C) concentrations were increased in patients with diabetes, however, no relation was found to catecholamine release. Diabetic and non-diabetic patients did not differ regarding left ventricular ejection fraction and arterial blood pressure. CONCLUSION/INTERPRETATION: Cardiac norepinephrine release is suppressed in patients with Type 2 diabetes which could contribute to sympathetic neuropathy. The difference of norepinephrine release in diabetic and non-diabetic patients was independent of cardiac function and arterial blood pressure. PMID- 12739024 TI - Apolipoprotein e4 allele and endothelium-dependent arterial dilation in Type 2 diabetes mellitus without angiopathy. AB - AIMS/HYPOTHESIS: Several studies have suggested a predisposing role of the e4 allele of apolipoprotein E (ApoE) in the development of atherosclerosis and cardiovascular disease in Type 2 diabetes. Therefore, we hypothesized that the e4 allele is also a risk factor for endothelial dysfunction. We attempted to assess whether Apo e4 allele is associated with endothelial dysfunction in the early stage of Type 2 diabetes. METHODS: We selected 255 Chinese Han Type 2 diabtetic men without angiopathy. PCR or allele-specific oligonucleotide probes were used to analyse ApoE genotypes, and high resolution ultrasound was used to measure brachial artery diameter at rest, after reactive hyperaemia and after sublingual glyceryltrinitrate. RESULTS: The flow-mediated arterial dilation among the subjects with e4/3 or e4/4 was 3.14+/-0.32%, which was lower than that in subjects with e2/2 or e3/2 (4.04+/-0.30%) ( p=0.038). The baseline vessel size, glyceryltrinitrate-induced arterial dilation and baseline flow were not different among different ApoE genotypes. On univariate analysis, reduced flow-mediated arterial dilation was related to total cholesterol, LDL, lipoprotein(a) [Lp(a)], high blood pressure, older age, family history of premature vascular disease, larger vessel size, cigarette smoking, duration of diabetes and e4 allele ( p<0.05). By multiple stepwise regression analysis, reduced flow-mediated arterial dilation was associated with cigarette smoking, LDL, Lp(a), and e4 allele ( p<0.01). CONCLUSION/INTERPRETATION: Apo e4 allele is associated with impairment of endothelium-dependent arterial dilation in the early stage of Type 2 diabetes. PMID- 12739026 TI - In situ protein Kinase C activity is increased in cultured fibroblasts from Type 1 diabetic patients with nephropathy. AB - AIMS/HYPOTHESIS: To verify whether individual susceptibility to diabetic nephropathy resides in an intrinsic difference in Protein Kinase C (PKC) activity. METHODS: We compared the effect of different glucose concentrations on PKC activity, PKC isoform expression and diacylglycerol (DAG) content in cultured fibroblasts from 14 Type 1 diabetic patients who developed nephropathy with those in cells from 14 patients without nephropathy. We recruited 14 normal subjects as control patients. Forearm skin fibroblasts were cultured in either normal (5 mmol/l) or high (20 mmol/l) glucose concentrations. RESULTS: In normal glucose, in situ PKC activity was higher in Type 1 patients with nephropathy (10.1+/-1.4 pmol/min/mg protein; p<0.01) than in those without (6.8+/-0.8) and the normal control subjects (6.3+/-0.5). This difference was due to increased concentrations of PKCalpha isoform in the membrane fraction of fibroblasts from patients with nephropathy. DAG content was also higher in cells from Type 1 patients with nephropathy. Incubation in high glucose concentration caused a further increase in PKC activity and DAG content in quiescent fibroblasts from patients with diabetic nephropathy, with no significant changes in cells from diabetic patients without nephropathy and normal control subjects. CONCLUSION/INTERPRETATION: Differences in PKC activation could contribute to the individual susceptibility to renal damage in Type 1 diabetic patients. PMID- 12739027 TI - Oxidant regulation of gene expression and neural tube development: Insights gained from diabetic pregnancy on molecular causes of neural tube defects. AB - AIMS/HYPOTHESIS: Maternal diabetes increases oxidative stress in embryos. Maternal diabetes also inhibits expression of embryonic genes, most notably, Pax 3, which is required for neural tube closure. Here we tested the hypothesis that oxidative stress inhibits expression of Pax-3, thereby providing a molecular basis for neural tube defects induced by diabetic pregnancy. METHODS: Maternal diabetes-induced oxidative stress was blocked with alpha-tocopherol (vitamin E), and oxidative stress was induced with the complex III electron transport inhibitor, antimycin A, using pregnant diabetic or non-diabetic mice, primary cultures of neurulating mouse embryo tissues, or differentiating P19 embryonal carcinoma cells. Pax-3 expression was assayed by quantitative RT-PCR, and neural tube defects were scored by visual inspection. Oxidation-induced DNA fragmentation in P19 cells was assayed by electrophoretic analysis. RESULTS: Maternal diabetes inhibited Pax-3 expression and increased neural tube defects, and alpha-tocopherol blocked these effects. In addition, induction of oxidative stress with antimycin A inhibited Pax-3 expression and increased neural tube defects. In cultured embryo tissues, high glucose-inhibited Pax-3 expression, and this effect was blocked by alpha-tocopherol and GSH-ethyl ester, and Pax-3 expression was inhibited by culture with antimycin A. In differentiating P19 cells, antimycin A inhibited Pax-3 induction but did not induce DNA strand breaks. CONCLUSION/INTERPRETATION: Oxidative stress inhibits expression of Pax-3, a gene that is essential for neural tube closure. Impaired expression of essential developmental control genes could be the central mechanism by which neural tube defects occur during diabetic pregnancy, as well as other sources of oxidative stress. PMID- 12739030 TI - Aspirin use is associated with higher serum concentrations of the anti-oxidant enzyme, paraoxonase-1. PMID- 12739029 TI - A genome scan for loci linked to quantitative insulin traits in persons without diabetes: the Framingham Offspring Study. AB - AIMS/HYPOTHESIS: Hyperinsulinaemia and insulin resistance usually precede clinical hyperglycaemia and Type 2 diabetes. Thus, plasma insulin concentrations and insulin resistance are important quantitative traits associated with risk of Type 2 diabetes, and represent key measures for genetic analysis of the syndrome. METHODS: We carried out a genome-wide search for loci related to plasma insulin concentrations and insulin resistance in 330 extended, community-based pedigrees from the Framingham Heart Study. Normalized deviates of the standardized residuals of plasma insulin concentrations in the fasting state, 2 h after oral glucose challenge and as a measure of insulin resistance were used in linkage analysis with the variance components model implemented in the computer program SOLAR. RESULTS: The results suggest susceptibility loci influencing plasma concentrations of fasting insulin and insulin resistance on chromosomes 11 (LOD 2.43 at 85 cM close to D11S2002) and 17 (LOD 1.8 at 60 cM, close to D17S784); and susceptibility loci influencing 2-h plasma insulin concentrations on chromosomes 9 (LOD 2.8 at 80 cM, close to D9S922) and 19 (LOD 1.8 at 66 cM, close to D19S245). The results of the analysis of 1000 simulations of the trait and an unlinked marker suggest that in a genome scan of 401 markers fewer than one LOD score over 1 would be due to Type 1 error, and be a false positive. CONCLUSION/INTERPRETATION: We conclude that these suggestive regions for quantitative pre-diabetic insulin traits could contain major loci in the pathogenesis of Type 2 diabetes. PMID- 12739028 TI - Early and intermediate Amadori glycosylation adducts, oxidative stress, and endothelial dysfunction in the streptozotocin-induced diabetic rats vasculature. AB - AIMS/HYPOTHESIS: In a model of streptozotocin-induced Type 1 diabetes mellitus in rats of 9 weeks duration, we analysed time associations between the development of hyperglycaemia, early and intermediate glycosylation Amadori adducts, or AGE compared with enhancement of oxidative stress and endothelial dysfunction. METHODS: Endothelial function was tested at several stages of streptozotocin induced diabetes and after treatment with insulin, resulting in different concentrations of blood glucose, glycosylated haemoglobin (an Amadori adduct), and AGE. Other animals were studied antagonising the formation of AGE with aminoguanidine. RESULTS: Relaxation in response to acetylcholine (1 nmol/l to 10 micro mol/l) was tested in isolated segments from aorta or mesenteric microvessels. Impairment of endothelium-dependent relaxations occurred after 2 weeks of untreated diabetes. Preincubation of vessels affected with 100 U/ml superoxide dismutase improved the relaxations to acetylcholine, along the time course of the endothelial impairment. This indicates the participation of reactive oxygen species on diabetic endothelial dysfunction. The impairment of endothelium-dependent relaxations was recovered after 3 more weeks of insulin treatment. Aminoguanidine treatment did not modify this pattern of development. The time course of the rise and disappearance of endothelial dysfunction showed a higher correlation with glycosylated haemoglobin concentrations than with blood glucose or serum AGE. CONCLUSION/INTERPRETATION: Enhancement of early and intermediate Amadori adducts of protein glycosylation was the factor showing a better relation with the development of endothelium impairment. These results are consistent with a role for these products in the development of diabetic vasculopathy. PMID- 12739031 TI - Bilateral synchronous tonsillar carcinoma in cervical cancer of unknown primary site (CUPS). AB - In addition to panendoscopy and imaging for staging, ipsilateral tonsillectomy is a standard procedure in the search for the primary tumor in cervical cancer of unknown primary site (CUPS). It is not clear from the literature, whether bilateral tonsillectomy has been established as the standard procedure in cancer of unknown primary origin. A bilateral synchronous tonsillar carcinoma has thus far only been described three times in the literature. We report on a case of CUPS in which a bilateral tonsillar carcinoma was detected after bilateral tonsillectomy. We also discuss the inclusion of bilateral tonsillectomy as a standard procedure in the search for primary malignancies. To diagnose and adequately treat a bilateral synchronous tonsillar carcinoma without losing time, we recommend bilateral tonsillectomy as a standard procedure in cervical CUPS. PMID- 12739032 TI - Expression of collagenase-1 (MMP-1), collagenase-3 (MMP-13) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in laryngeal squamous cell carcinomas. AB - Matrix metalloproteinases (MMPs) that have a proteolytic activity against the components of extracellular matrix (ECM) play an important role in the invasive and metastatic spread of tumors. The role of MMPs and tissue inhibitors of MMPs (TIMPs) in laryngeal squamous cell carcinoma (LSCC) has not been elucidated sufficiently. The aim of the present study was to evaluate the correlation between the expression of collagenase-1 (MMP-1), collagenase-3 (MMP-13) and TIMP 1, as well as the clinicopathological features of LSCCs. The expression of collagenases and TIMP-1 was examined immunohistochemically in 50 cases of surgically obtained specimens of primary LSCCs. Analyses indicated that LSCC cells as well as stromal cells expressed MMP-1, MMP-13 and TIMP-1 immunostaining. Overexpression of TIMP-1 occurred more frequently in non-metastasizing cases (P=0.009). TIMP-1 and MMP-1 staining correlated significantly with the histologic type of LSCC. The keratinizing type of carcinomas exhibited higher TIMP-1 protein expression than the nonkeratinizing variety (P=0.01). TIMP-1 staining was associated with the grade of differentiation, since it was found predominantly in well and moderately differentiated carcinomas (P=0.04). The findings confirm that expression of analyzed MMPs and TIMP-1 is characteristic of LSCC and that these enzymes contribute to the progression of tumors. TIMP-1 upregulation might exhibit lower metastatic potential in LSCCs and is linked rather with an early stage of tumor progression. It seems also that TIMP-1 expression is dependent on the grade of differentiation. PMID- 12739033 TI - Transient marrow edema syndrome of the hip: results after core decompression. A prospective MRI-controlled study in 22 patients. AB - BACKGROUND: Transient marrow edema syndrome (TMES) of the hip is a disease of acute onset and severe functional disability. There is histological evidence for an ischemic etiology of TMES of the hip. Core decompression as applied for avascular necrosis (AVN) of the hip is therefore a therapeutic alternative to conservative therapy, the latter leading only to a reduction of symptoms but never a shortening of the course of the disease. METHODS: Between January 1998 and June 2000, 22 hips with TMES were treated with core decompression in our department. TMES was diagnosed by exclusion. MRI was done preoperatively and at 6 months postoperatively. The postoperative MRI result was classified into three categories. RESULTS: After an average of 7.2 (range 1-30) days, all patients were pain-free after core decompression. In 2 patients, TMES progressed to AVN despite core decompression. All others had no signal alterations of the head of the femur on MRI after 6 months. The postoperative Harris Hip Score (HHS) in patients with TMES was on average 93.7 (range 77-95); in patients with AVN, the postoperative HHS was 47 (range 45-49). CONCLUSION: Our results demonstrate that core decompression of the hip significantly shortens the natural course of disease of TMES of the hip. PMID- 12739034 TI - Distal foregut atresias in consecutive siblings and twins in the same family. AB - The distal foregut comprises the antrum of the stomach and supra-ampullary region of the duodenum. In this part of the gut, intraluminal mucosal diaphragms, webs, membranes (type I), and rarely solid cord (type II) atresias are occasionally seen in clinical practice. Due to increased awareness, the number of reported cases has increased in recent years. These foregut lesions are known to have a strong genetic association. Both antral (AA) and duodenal atresias (DA) may occur as potentially curable sporadic cases or in combination with lethal malformations such as hereditary multiple gastrointestinal atresias affecting the fore-, mid-, and hindgut. The association of AA and junctional epidermolysis bullosa lethalis is well-known. We describe an unusual occurrence of foregut atresias in two consecutive siblings and a pair of monozygous twins. The involvement of four siblings in a single family is extraordinary and as yet unreported. This cluster of cases not only lends a strong support to a genetic origin, but also the mode of transmission, despite the absence of a history of consanguinity. It indicates that atresia of the gastric antrum and supra-ampullary DA are etiologically closely related. It is also possible that the discordant expression in the twins could be a pleiotropic effect of the same genetic factor. Attempts are made to explain the possible pathogenesis of this unusual atresia and relate it to other associated anomalies. PMID- 12739035 TI - Frontolateral keyhole craniotomy through a superciliary skin incision in children. AB - INTRODUCTION: In our experience, frontolateral keyhole craniotomy in children is a safe approach for an experienced neurosurgeon to use in the treatment of the tumors or arachnoid cysts of the anterior fossa and sellar regions. TECHNICAL NOTE: The authors describe the frontolateral keyhole craniotomy, which is a modification of the generally used pterional approach. The operations were carried out through an approximately 2.5 x 3-cm frontolateral miniaturized craniotomy after a skin incision just above the eyebrow. We adopted this technique in pediatric neurosurgery. PMID- 12739036 TI - Differential diagnosis of high serum creatine kinase levels in systemic lupus erythematosus. AB - We report the clinical and bioptic findings for a 57-year-old woman with severe chloroquine-induced myopathy. Since 1989, she had been suffering from systemic lupus erythematosus (SLE) with renal involvement and undergone periods of treatment with azathioprine and cyclophosphamide. Additional therapy with chloroquine (CQ) was started because of arthralgia. At the same time, slightly increased creatine kinase (CK) levels were noted. Myositis was suspected, and the patient was treated with steroids. The CK increase persisted, however, and she developed progressive muscular weakness and muscular atrophy. Routine controls revealed markedly elevated CK levels of 1,700 U/l. The neurological and electrophysiological findings were not typical of myositis. Thus, muscle biopsy of the deltoid muscle was performed in order to exclude polymyositis or toxic myopathy. As it revealed chloroquine-induced myopathy, medication was stopped. Discriminating between primary SLE-induced affection of the musculoskeletal system and drug-induced side effects is important for appropriate treatment of SLE patients. PMID- 12739038 TI - Significance of catechol-O-methyltransferase gene polymorphism in fibromyalgia syndrome. AB - Fibromyalgia syndrome (FS) is associated with a neuroendocrinal disorder characterized by abnormal function of the hypothalamic-pituitary-adrenal (HPA) axis, including hyperactive adrenocorticotropic hormone (ACTH) release and adrenal hyporesponsiveness. Catechol-O-methyltransferase (COMT) enzyme inactivates catecholamines and catecholamine-containing drugs. Polymorphism in the gene encodes for the COMT enzyme. For this study, the significance of COMT polymorphism was assessed in FS. There were three polymorphisms of the COMT gene: LL, LH, and HH. The analysis of COMT polymorphism was performed using polymerase chain reaction (PCR). Sixty-one patients with FS and 61 healthy volunteers were included in the study. Although no significant difference was found between LL and LH separately, the LL and LH genotypes together were more highly represented in patients than controls ( P=0.024). In addition, HH genotypes in patients were significantly lower than in the control groups ( P=0.04). There was no significant difference between COMT polymorphism and psychiatric status of the patients as assessed by several psychiatric tests ( P>0.05). In conclusion, COMT polymorphism is of potential pharmacological importance regarding individual differences in the metabolism of catechol drugs and may also be involved in the pathogenesis and treatment of FS through adrenergic mechanisms as well as genetic predisposition to FS. PMID- 12739039 TI - The biallelic variable number of tandem repeats of the tumor necrosis factor receptor 2 promoter in systemic lupus erythematosus. AB - The objective of this study was to investigate whether the variable number of tandem repeats (VNTR) of the tumor necrosis factor receptor 2 (TNFR2) promoter is associated with susceptibility to systemic lupus erythematosus (SLE) and its clinical phenotypes. A biallelic VNTR within a 42-bp region in the TNFR2 gene promoter was determined by polymerase chain reaction in 88 SLE patients and 95 healthy control subjects. Clinical manifestations were analyzed in each patient and correlated with the genotypes. When the TNFR2 promoter VNTR was compared between Korean and Caucasian healthy controls with respect to allele frequencies, there was a significant difference (alleles 1 and 2: 39, 151 in Koreans vs 60, 138 in Caucasians, respectively; chi-squared test 4.38; 2 df; P=0.036). The genotype distribution of the TNFR2 promoter VNTR did not differ between SLE patients and control subjects (1.1, 1.2, and 2.2 genotypes 7, 14, 67 vs 5, 29, and 61 controls, respectively; chi-squared test 5.19; 2 df; P=0.061). According to the TNFR2 promoter genotypes in the lupus patients, clinically there was no significant difference in age at onset, anti-dsDNA titer, C4 level, systemic lupus erythematosus disease activity index (SLEDAI), Systemic Lupus International Collaborating Clinics/American College of Rheumatology (SLICC/ACR) damage index, or autoantibodies. However, the 1.1 genotype group showed the lowest C3 level and more frequent renal involvement than the 1.2 and 2.2 genotype groups. In conclusion, an ethnic difference in the TNFR2 promoter VNTR has been found and the biallelic VNTR of the TNFR2 promoter may be associated with clinical phenotypes in SLE. PMID- 12739040 TI - Ex vivo apoptosis, CD95 and CD28 expression in T cells of children with juvenile idiopathic arthritis. AB - We hypothesise that T-cell apoptosis and the percentage of T cells expressing molecules involved in apoptosis modulation (CD95, CD28) are altered at the inflammation site and in peripheral blood (PB) of children with juvenile idiopathic arthritis (JIA). Paired JIA samples of synovial fluid (SF) and PB ( n=7) and PB samples from age-matched normal children ( n=23) were analysed immediately ex vivo. Apoptosis was measured by detection of phophatidylserine (PS) externalisation on T cells. CD95 or CD28 was detected by FACS, and soluble CD95 and CD95 ligand levels were detected by enzyme-linked immunosorbent assay (ELISA). In SF, the mean percentage of apoptotic T cells was somewhat higher than in PB. However, the percentages of T cells expressing CD95 and soluble CD95 levels were markedly higher in SF (CD4 cells 96+/-2%, CD8 91+/-6%, soluble CD95 6,420+/-2,571 pg/ml) than in PB (CD4 32+/-10%, CD8 36+/-9%, soluble CD95 4,296+/ 2,142 pg/ml). Peripheral blood T-cell apoptosis in JIA (CD4 20+/-8%, CD8 42+/ 19%) was higher than in the control group (CD4 5+/-2%, CD8 9+/-6%). Interestingly, the percentage of PB CD4 cells expressing CD28 was lower in JIA than controls. In conclusion, systemic T-cell apoptosis was higher in JIA while a substantial number of SF T cells survived locally, despite the fact that almost all cells express CD95. PMID- 12739037 TI - Polymorphism of the beta3-adrenergic receptor and lipid profile in patients with rheumatoid arthritis and systemic lupus erythematosus treated with chloroquine. AB - We investigated the effect of beta 3-adrenergic receptor (beta(3)AR) polymorphism on lipid profiles in patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) treated with chloroquine. One hundred sixty-eight subjects were classified into three groups: 61 RA patients, 57 SLE patients, and 50 healthy subjects. All patients fulfilled the 1987 and 1982 classification criteria for RA and SLE, respectively, of the American College of Rheumatology. Demographic data and clinical characteristics of the patients were registered. Fasting lipid profile determination and leukocyte genomic DNA isolation from peripheral blood was performed in all the participants. Screening of the beta(3) AR gene polymorphic region (exon 1) was done by the polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) technique. Quantitative and qualitative variables were analyzed using analysis of variance (ANOVA) with the LSD and chi(2) tests, respectively. An association between the arg64/arg64 beta(3)-AR genotype and high levels of triglycerides (TG) and very low-density lipoprotein cholesterol (VLDL-c) was found in three RA patients ( P=0.01), two of them taking chloroquine. Arg64/arg64 beta(3)-AR polymorphism may contribute to increased TG and VLDL-c in RA patients, independently of chloroquine treatment. PMID- 12739041 TI - Polymorphism of the Duffy erythrocyte chemokine receptor in Italian patients with Behcet's disease. AB - This study examined the hypothesis that the polymorphism of Duffy antigen receptor for chemokines (DARC) predisposes to and/or influences the clinical manifestations of Behcet's disease. The serum levels of IL-8 and monocyte chemotactic peptide (MCP)-1, two DARC-binding chemokines, were investigated and related to this polymorphism. Twenty-eight patients with Behcet's disease and 30 healthy blood donors were included in the study. No null phenotypes were found among the patients studied, and the frequencies of the other phenotypes (Fy((a+b )), Fy((a+b+)), and Fy((a-b+))) did not significantly differ from those found in the blood donor group or reported in the general Caucasian population. No difference was found between the single phenotypes in terms of IL-8 and MCP-1 serum levels, and no relevant association between the clinical characteristics, Behcet's disease-associated human leukocyte antigen (HLA)-B51, and single phenotypes was observed. This investigation indicates that DARC is not a genetic trait significantly associated with or predisposing to Behcet's disease, at least in Caucasian Italians. However, the role of this polymorphism in the development and in the clinical course of the disease awaits further investigation. PMID- 12739042 TI - Functional disability and quality of life in patients with ankylosing spondylitis. AB - The aim of this study was to evaluate functional disability and quality of life (QOL) in patients with ankylosing spondylitis (AS) and determine the relationship between functional status and measures of clinical condition including QOL. Fifty one AS patients (45 male, six female) with a mean age of 37.2+/-10.8 years were included. The demographic data of the patients were recorded. Their clinical status was assessed using the Bath Ankylosing Spondylitis Metrology Index (BASMI), Bath Ankylosing Spondylitis Radiology Index (BASRI), and Bath Ankylosing Spondylitis Disease Activity Index (BASDAI). Global pain of the patients was determined with a visual analog scale (VAS), and QOL status was evaluated with the Nottingham Health Profile (NHP). Twenty-seven patients (52.9%) had peripheral articular involvement. Sixty percent had mild-to-moderate and 25.4% of the patients had severe functional disability, while 5.8% did not report any functional loss. A significant change in the mean scores of all clinical measures except BASRI was observed between patients with and without peripheral arthritis. The clinical measures of disease (BASRI, BASMI, and BASDAI) were all correlated with each other and with laboratory variables. The strongest factors correlating with functional loss were BASMI and BASDAI. The scores of all sections of the NHP were significantly higher, indicating a poor quality of life in AS patients. Peripheral joint involvement had a significant role in the deterioration of QOL. Physical domains of NHP such as pain and physical activity had highest correlations with functional disability, whereas psychosocial domains of NHP were found to correlate more highly with BASDAI and VAS pain scores. These results show the effect of AS, especially when the disease is active and associated with peripheral involvement. In conclusion, current management strategies should focus on decreasing pain, maintaining physical activity, and efforts to improve psychosocial health aspects for increasing QOL in patients suffering from AS. PMID- 12739044 TI - Plasma thrombomodulin levels in patients with Behcet's disease. AB - The plasma levels of thrombomodulin in 54 patients with Behcet's disease (BD) and 20 healthy control subjects were studied. The mean thrombomodulin (TM) level was significantly higher in active BD patients than in inactive patients and healthy controls ( P<0.001 and <0.01, respectively). Plasma TM levels did not show a significant relation with clinical manifestations. Increased plasma TM levels are associated with active disease and may reflect the presence of endothelial cell activation and/or injury. PMID- 12739043 TI - Factors playing a role in the development of decreased bone mineral density in juvenile chronic arthritis. AB - OBJECTIVE: The aims of this study were to evaluate bone mineral density (BMD) in patients with juvenile chronic arthritis (JCA), compare them with healthy controls, and assess the effects of disease activity and corticosteroid treatment on BMD. METHODS: Twenty-eight patients diagnosed with JCA and 45 healthy controls were included in this study. Disease activity was determined by clinical and laboratory evaluation, Articular Disease Severity Score (ADSS), and the Juvenile Arthritis Functional Assessment Report (JAFAR). Bone mineral density of the lumbar spine was measured by dual energy X-ray absorptiometry (DEXA). RESULTS: Patients with JCA showed significant decreases in BMD compared with healthy controls. The JCA patients treated with corticosteroids showed significantly lower BMDs than the healthy control group. Age of the patients and age of onset were found to correlate with BMD. CONCLUSION: Our study showed that glucocorticoids were involved in the development of osteoporosis in JCA, with many other factors affecting bone mineralization. We could not demonstrate any relationship between BMD and disease activity, but the study data suggest that early onset disease is also an important factor in the development of osteoporosis in JCA. PMID- 12739046 TI - Peripheral T cell lymphoma-associated hemophagocytic syndrome in a patient with human adjuvant disease. AB - Hemophagocytic syndrome (HPS) is a rare complication of lymphoma. We report a 70 year-old woman with human adjuvant disease who developed lymphoma-associated HPS (LAHPS) and elevated serum soluble interleukin (IL)-2 receptor. Despite intensive therapy, she died of pneumonia. Necropsy revealed a prominent spleen containing atypical T cells and many erythrophagocytizing histiocytes. PMID- 12739045 TI - Methotrexate inhibits interleukin-6 production in patients with juvenile rheumatoid arthritis. AB - OBJECTIVES: Methotrexate (MTX) is one of the most widely used disease-modifying antirheumatoid drugs in the treatment of juvenile rheumatoid arthritis (JRA). We studied its effect on the production of two proinflammatory cytokines, interleukin-6 (IL-6) and tumor necrosis factor alpha (TNFalpha), by peripheral blood cells in patients with JRA. METHODS: Interleukin-6 and TNFalpha levels were measured at 0 and 4 weeks in whole blood cultures with and without lipopolysaccharide (LPS) stimulation in 19 children treated with MTX (10 mg/m(2 )per week) or placebo. Ten healthy individuals were included as healthy controls. RESULTS: Spontaneous production of IL-6 and TNFalpha by peripheral blood cells of patients with JRA was higher than in healthy controls ( P<0.01). However, IL-6 and TNFalpha production after LPS stimulation was similar in healthy controls and patients. The two groups of patients, i.e., those treated with placebo and those treated with MTX, had similar spontaneous and induced IL-6 and TNFalpha production. At 4 weeks, the drop in spontaneous IL-6 and TNFalpha production was no different in the two groups, but LPS-stimulated IL-6 production was significantly lower in the MTX-treated group than the placebo group ( P<0.05). CONCLUSION: Methotrexate reduces the production of IL-6 by activated cells, and this may be responsible for its anti-inflammatory property. PMID- 12739047 TI - A challenging case of rheumatoid arthritis in an acromegalic patient. AB - A 63-year-old man with complaints of joint pain and ankle swelling was evaluated. The arthralgias he described were mainly in the knees, elbows, and shoulders. Accompanying swelling and erythema in his left ankle and left second metacarpophalangeal (MCP) joint had recently ensued. His past history revealed acromegaly, somatotropinectomy, and radiotherapy. His neck, bilateral wrist, elbow, and shoulder joints were involved; there was pain and limited range of motion. The MCP joints, being worse than the interphalangeal joints, were likewise involved. His left ankle and MCP joints additionally were swollen and erythematous. Laboratory and radiological evaluations were carried out. Radiological and clinical findings confirmed a diagnosis of rheumatoid arthritis and concurrent acromegalic arthropathy. The patient was treated accordingly. Interestingly, he later developed colon cancer. PMID- 12739048 TI - Juvenile chronic arthritis in a monozygotic twin couple. PMID- 12739050 TI - [Vasculitides: classification, clinical aspects and pathology. A review]. AB - The Consensus Conference of Chapel Hill (1994) laid a common base for a standardized classification of vasculitides which, after a pre-existing historical diversity, enabled a comparison of different studies. The criteria for this classification are foremost anatomico-pathological and are based on the caliber of affected blood vessels. In addition the quality of the inflammatory reaction, and the entirety of clinical symptoms, as well as the results of immunological tests are included. Moreover a unified pathogenetic concept is proposed for all types of vasculitis. The diagnosis of vasculitis of large, medium or small vessels is made primarily by biopsies and examination of HE stained sections. However, overlap of calibers of vessels affected and lack of specificity of inflammatory reaction with individual vasculitis types have to be paid attention to. Immunohistochemical methods are only rarely routinely applied. In contrast the complete clinical picture and the results of immunological laboratory tests (especially AntiNeutrophilCytoplasmicAntibodies, ANCA) play an important role in the differential diagnosis of various vasculitis types. Definitions, clinical findings and pathology of the various types of vasculitis are described. Also mentioned are secondary vasculitides, e.g concurrent to collagen diseases or infections. Not only the diagnosis of vasculitis but also the recognition of the specific type and, last but not least monitoring by histological controls, may be decisive for an optimal therapy. PMID- 12739049 TI - Genetic conservation versus variability in mitochondria: the architecture of the mitochondrial genome in the petite-negative yeast Schizosaccharomyces pombe. AB - The great amount of molecular information and the many molecular genetic techniques available make Schizosaccharomyces pombe an ideal model eukaryote, complementary to the budding yeast Saccharomyces cerevisiae. In particular, mechanisms involved in mitochiondrial (mt) biogenesis in fission yeast are more similar to higher eukaryotes than to budding yeast. In this review, recent findings on mt morphogenesis, DNA replication and gene expression in this model organism are summarised. A second aspect is the organisation of the mt genome in fission yeast. On the one hand, fission yeast has a strong tendency to maintain mtDNA intact; and, on the other hand, the mt genomes of naturally occurring strains show a great variability. Therefore, the molecular mechanisms behind the susceptibility to mutations in the mtDNA and the mechanisms that promote sequence variations during the evolution of the genome in fission yeast mitochondria are discussed. PMID- 12739051 TI - [Gastrointestinal stromal tumors. A morphologic and molecular genetic independent tumor entity with new therapeutic perspectives]. AB - Recent morphological and molecular genetic findings have greatly expanded our understanding of gastrointestinal stromal tumors (GISTs). GISTs are now defined by their overexpression of CD117 (KIT), the receptor for the stem cell factor, and can thus be discriminated from smooth muscle tumors. Cytogenetically, GISTs are characterized even in early lesions by frequent entire or partial loss of the chromosomes 14 and 22 and terminal deletions of the chromosomal arm 1p. During tumor progression further chromosomal imbalances accumulate. Following the first report on activating KIT mutations in GISTs, several studies have addressed the role of wild-type and mutant KIT in GISTs and demonstrated activating KIT mutations in the majority of cases. Moreover, KIT tyrosine phosphorylation is even present in KIT mutation-negative GISTs, implicating KIT activation as a central event in the pathogenesis of GISTs. Imatinib (STI571/Glivec) is a selective inhibitor of BCR/ABL, PDGFR and KIT receptor-tyrosine kinases. First therapeutic applications of imatinib in patients with progressive GISTs have yielded promising results. This review focuses on the morphological and molecular findings in GISTs which have opened up a new therapeutic perspective. PMID- 12739052 TI - [Human intestinal spirochetosis]. AB - Whenever biopsy material obtained from endoscopically normal colorectal mucosa reveals the blue haematoxyphilic line between the microvilli of the covering epithelium, the rare condition of intestinal spirochetosis is diagnosed. The classification of the bacteria detected with the aid of special stains (e.g. the Warthin Starry silver stain) and in the electron microscope, continues to be something of a problem. A further point of contention is the question whether this spirochetal infection is of pathological significance or not. A point mitigating against pathogenicity is the fact that no histological signs of an inflammatory reaction are to be seen. Also, the symptoms of patients with intestinal spirochetosis are such that they provide no basis for a pronouncement on whether the infection is of a pathological or a pathological nature. On the other hand, however, a number of studies do seem to indicate that the spirochetes might be the cause of such symptoms as diarrhoea, constipation and abdominal pain. A point that would appear to support this view is the fact that such symptoms may disappear after successful treatment with metronidazole. The histological diagnosis is easily established when, faced by an apparently normal histological appearance of the colorectal mucosa, the pathologist considers the possibility of spirochetosis, and undertakes a specific search for the blue haematoxyphilic line in the covering epithelium of the colorectal mucosa. PMID- 12739054 TI - [The autopsy law of Hamburg from February 9, 2000--changing autopsy rates in departments of pathology in Hamburg]. AB - The decline of the autopsy rate in institutes for pathology in Germany is a reason for growing concern. One reason for the restrained activity in performing autopsies might be uncertainty about the legal position concerning the admissibility of so-called clinical or scientific autopsies. Whereas a legal basis for different kinds of autopsies does exist for quite some time (i.e. code of criminal procedure, epidemic law etc.), such statutory rulings have been lacking for clinical or scientific autopsies. Following Berlin and Bremen, in 2000 Hamburg introduced a law for the regulation of scientific autopsies. This autopsy law differentiates between clinical, medicolegal (not on public prosecutor's order), and anatomical autopsies. It arranges aspects of legality of, consent in, order of, and implementation of the autopsy. A survey among clinical pathologists in Hamburg clarifies the drop in the autopsy rate since the last decades until today. The colleagues were also asked to point out their opinion about the reasons for the dropping autopsy rate and to state their experience with the new autopsy law. PMID- 12739053 TI - [Pathogenesis and histomorphology of ampullary carcinomas and their precursor lesions. Review and individual findings]. AB - Most adenomas and carcinomas of small intestine and extrahepatic bile ducts arise in the region of Vater's papilla. In FAP it is the main location for carcinomas after proctocolectomy. In many cases symptoms due to stenosis lead to diagnosis in an early tumor stage. In about 80% curative intended resection is possible. Operability is the most relevant prognostic factor. Inflammatory changes, fibrosis, regeneratory changes after endoscopic manipulation, hyperplasia, preneoplastic lesions close to carcinoma, deeply sited carcinomas under protruded, non-neoplastic duodenal mucosa make the diagnosis difficult on biopsy material. Histologically, intestinal type adenocarcinoma, pancreatobiliary type adenocarcinoma, undifferentiated carcinomas and unusual types can be differentiated. In our own series comprising 45 resected ampullary carcinomas 6 from 10 intestinal type adenocarcinomas, and 4 carcinomas of unusual types expressed the immunohistochemical marker profile of intestinal mucosa (keratin 7 , keratin 20+, MUC2+). 17 from 21 pancreatobiliary type adenocarcinomas, 4 undifferentiated carcinomas, as well as 3 papillary carcinomas showed the immunohistochemical profile of pancreaticobiliary duct mucosa (keratin 7+, keratin 20-, MUC2-). 3 invasive carcinomas which were negative for these markers, showed one of these characteristic marker-combinations in non-invasive adenomatous parts. These findings support the concept of histogenetically different ampullary carcinomas which are developing from the intestinal or from the pancreaticobiliary type mucosa of Vater's papilla. Molecular alterations in ampullary carcinomas are similar to those of colorectal as well as pancreatic carcinomas, although they appear in different frequencies. In future studies molecular alterations in ampullary carcinomas should be correlated closely with the different histologic tumor types. The histologic classification should reflect consequently the histogenesis of ampullary tumors from the two different types of papillary mucosa. PMID- 12739055 TI - [Cutaneous perineurioma. Clinical and histological findings and differential diagnosis]. AB - Perineuriomas, composed almost exclusively of EMA (epithelial membrane antigen) positive cells, represent a rare and distinct entity in the spectrum of nerve sheath tumours. At present three subtypes, including intraneural and extraneural perineurioma as well as sclerosing perineurioma, can be distinguished; atypical and malignant perineuriomas are extremely uncommon. We analysed the clinicopathological and immunohistochemcial features of 13 cases of cutaneous perineurioma. The neoplasms arose in adult patients (age range from 18 to 71 years) and were seen on the hand (six cases), the thigh (three cases), the lower leg (two cases), the forearm (one case) and in perinasal location (one case). Morphologically, seven neoplasms showed features of dermatofibroma-like perineurioma, four lesions were consistent with sclerosing perineurioma, one cellular lesion resembled a solitary fibrous tumour, and one case was diagnosed as atypical/malignant cutaneous perineurioma; no local recurrences or tumour progression has been reported so far. The differential diagnosis of various variants of cutaneous perineurioma from other mesenchymal lesions as well as melanocytic and epithelial neoplasms is discussed. PMID- 12739056 TI - [Apoptosis and tumor regression in locally advanced non-small cell lung cancer with neoadjuvant therapy]. AB - Dysregulation of apoptosis is closely associated with malignant cell transformation. On the other hand, apoptosis is induced by chemotherapy or irradiation. Therefore, in 54 patients with locally advanced non-small cell lung cancer (NSCLC, 36 squamous cell carcinomas, 18 adenocarcinomas, stage IIIA/IIIB), apoptotic indices were comparatively analysed before onset and after termination of neoadjuvant therapy. The results were compared with the response to neoadjuvant therapy (extent of therapy-induced tumour regression) as well as the survival times. A statistically significant difference could not be established between pre-therapeutically and post-surgically established apoptotic indices (mean values: 0.93% vs. 1.1%). Neither before therapy nor after surgery did the apoptotic indices show a significant predictive value concerning different overall survival times. These results suggest that neoadjuvant therapy does not modify the extent of apoptosis in lung cancer in the long term. Only a few weeks after the completion of the neoadjuvant chemoradiotherapy this contributes to a net proliferation of the residual tumour tissue which is largely equivalent to that of the untreated tumour. PMID- 12739057 TI - [Rare mesenchymal lesions in siblings. Two case reports]. AB - We report on a 15-month-old boy presenting a juvenile active ossifying fibroma in the right nasal cavity and the sibling, a 9-month-old girl with a mesenchymal hamartoma of the chest wall. The two lesions showed many similarities. Both lesions are present at the time of birth or in early life with local obstructive or compressive effects. The lesions have a similar mixture of mature and immature mesenchymal tissue with areas of ossification. The entities present a tumor-like development with an abnormal mixture of tissue indigenous to the specific area of the body without notable atypical cytologic features. These features are typical criteria for hamartoma lesions. PMID- 12739059 TI - [Medicine in the land of stones]. PMID- 12739058 TI - [Primary malignant melanoma of the uterine cervix. Report of two cases and review of the literature]. AB - The manifestation of a malignant melanoma in the uterus is very rare, more often it is the result of metastasis rather than a primary tumor. A malignant melanoma at this site can originate either from melanocytic elements within the cervical epithelium or from the cervical stroma. We report on two cases of primary malignant melanoma of the uterine cervix and compare them with other cases from the literature. PMID- 12739060 TI - Pharmacokinetics of S-1, an oral formulation of ftorafur, oxonic acid and 5 chloro-2,4-dihydroxypyridine (molar ratio 1:0.4:1) in patients with solid tumors. AB - S-1 is an oral formulation of ftorafur (FT), oxonic acid and 5-chloro-2,4 dihydroxypyridine (CDHP) at a molar ratio of 1:0.4:1. FT is a 5-fluorouracil (5 FU) prodrug, CDHP is a dihydropyrimidine dehydrogenase (DPD) inhibitor and oxonic acid is an inhibitor of 5-FU phosphoribosylation in the gastrointestinal mucosa and was included to prevent gastrointestinal toxicity. We determined the pharmacokinetics of S-1 in 28 patients at doses of 25, 35, 40 and 45 mg/m(2). The plasma C(max) values of FT, 5-FU, oxonic acid and CDHP increased dose-dependently and after 1-2 h were in the ranges 5.8-13 microM, 0.4-2.4 microM, 0.026-1.337 microM, and 1.1-3.6 microM, respectively. Uracil levels, indicative of DPD inhibition, also increased dose-dependently from basal levels of 0.03-0.25 microM to 3.6-9.4 microM after 2-4 h, and 0.09-0.9 microM was still present after 24 h. The pharmacokinetics of CDHP and uracil were linear over the dose range. The areas under the plasma concentration curves (AUC) for CDHP and uracil were in the ranges 418-1735 and 2281-8627 micromol x min/l, respectively. The t(1/2) values were in the ranges 213-692 and 216-354 min, respectively. Cumulative urinary excretion of FT was predominantly as 5-FU and was 2.2-11.9%; the urinary excretion of both fluoro-beta-alanine and uracil was generally maximal between 6 and 18 h. During 28-day courses with twice-daily S-1 administration, 5-FU and uracil generally increased. Before each intake of S-1, 5-FU varied between 0.5 and 1 microM and uracil was in the micromolar range (up to 7 microM), indicating that effective DPD inhibition was maintained during the course. In a biopsy of an esophageal adenocarcinoma metastasis that had regressed, thymidylate synthase, the target of 5-FU, was inhibited 50%, but increased four- to tenfold after relapse in subsequent biopsies. In conclusion, oral S-1 administration resulted in prolonged exposure to micromolar 5-FU concentrations due to DPD inhibition, and the decrease in uracil levels after 6 h followed the pattern of CDHP and indicates reversible DPD inhibition. PMID- 12739061 TI - Pharmacokinetics and tissue distribution of cryptophycin 52 (C-52) epoxide and cryptophycin 55 (C-55) chlorohydrin in mice with subcutaneous tumors. AB - PURPOSE: To compare the pharmacokinetics and tissue distribution (both normal and tumor) of cryptophycin 52 (C-52) and its putative chlorohydrin prodrug cryptophycin 55 (C-55) in a murine model and to investigate a possible mechanism behind the superior activity of C-55. METHODS: Mammary adenocarcinoma 16/c tumor bearing mice were treated with an i.v. bolus of 11 mg/kg C-52 or 38 mg/kg C-55 in Cremophor-alcohol. At predetermined time intervals, C-52 and C-55 concentrations in plasma, liver, kidney, small intestine and tumors were measured using a previously described HPLC method. Pharmacokinetic parameters were computed using noncompartmental methods. Tissue (both normal and tumor) to plasma ratios as a function of time were also calculated for comparison. RESULTS: Both C-52 and C-55 were rapidly distributed into different tissues including tumors following i.v. administration. However, the affinities of these compounds towards different tissues were different. Thus, the half-lives (minutes) of C-55 were in the decreasing order liver (725), intestine (494), tumor (206), kidney (62) and plasma (44), whereas the AUC values (microg x min/ml) were in the order tumor (9077), liver (7734), kidney (6790), plasma (2372) and intestine (2234). For C 52, the half-lives (minutes) were in the decreasing order liver (1333), kidney (718), intestine (389), tumor (181) and plasma (35), and the AUC values (microg x min/ml) were in the order kidney (1164), liver (609), intestine (487), plasma (457) and tumor (442). The relative exposures to C-52 after i.v. injection of C 55 were plasma 3.9%, tumor 80.8%, kidney 3.4%, liver 1.1% and intestine 2.8%. Although plasma exposure to C-52 following C-55 administration was relatively small, the use of C-55 to deliver C-52 increased the retention of C-52 and its AUC in tumor compared to direct injection of C-52. Simultaneously, this approach shortened C-52 retention in all normal tissues studied. CONCLUSIONS: The distribution of C-55 and its bioconversion to C-52 in different organs and tumor tissue observed in this study suggest the ability of C-55 to target tumor tissue, creating a depot of C-52 in tumor. Increased C-52 exposure of tumor, with concomitant decreased exposure of normal tissue, is a contributing factor to the superior activity of C-55 versus C-52. However, except in the case of tumor tissue in which 81% of C-55 converts to C-52, only a minor amount of C-55 may serve as a prodrug for C-52, whereas the majority is handled by the biosystem through a different route of elimination. Tissue distribution combined with rate of conversion may be an important determinant of the relative effectiveness of other epoxide-chlorohydrin pairs of cryptophycins. PMID- 12739062 TI - Cellular and humoral immune reconstitution after autologous peripheral blood stem cell transplantation (PBSCT). AB - Immune reconstitution after autologous peripheral blood stem cell transplantation (PBSCT) is of particular interest because of its importance for clinical outcome. Despite prolonged immunosuppression, especially of CD4(+) cells, few infections after neutrophil recovery occur. Only reactivation of varicella zoster virus (VZV) is more frequent in the first year after transplantation. From August 1997 to May 2001, we prospectively evaluated 38 patients prior to conditioning and during follow-up of 12 months post-transplant for virus antibodies [measles, mumps, rubella, polio, herpes simplex, varicella zoster, mononucleosis, cytomegalovirus (CMV)] and lymphocyte subpopulations by flow cytometry. CD3(+) T lymphocytes, CD8(+) T cells, and B-lymphocyte reconstitution in our study confirms previous reports. Complete CD4(+) lymphocyte reconstitution was not achieved in the 12 months post-transplant leading to a suppressed CD4/CD8 ratio. IgG antibody titers against measles, mumps, rubella, and polio were present in almost all patients pretransplant and during 12 months post-transplant, indicating persistent humoral immunity. CD3(+) and CD8(+) counts of patients with clinical VZV reactivation ( n=5) post-transplant were significantly higher (median: 1201/microl and 938/microl, respectively) than in patients without VZV reactivation (median: 594/microl and 482/microl, respectively) 6-12 months post transplant. Positive CMV titers pretransplant ( n=19) were also correlated with higher CD3(+) and CD8(+) counts 3-6 months post-transplant (median: 1050/microl and 1056/microl, respectively) compared to CMV-negative patients (738/microl and 584/microl, respectively), although none of the patients suffered from CMV disease. Therefore, we conclude that persistent viral infections can contribute to the CD8(+) T-cell reconstitution after PBSCT by oligoclonal expansion of antigen-specific memory CD8(+) T cells. PMID- 12739063 TI - Circulating myeloid colony-forming cells predict survival in myelodysplastic syndromes. AB - The growth characteristics and the prognostic value of cytokine-stimulated myeloid colony formation from peripheral blood mononuclear cells (PBMC) of patients with myelodysplastic syndromes (MDS) are largely unknown. In this study we have determined the number of myeloid colony-forming units (mCFUs) in semisolid medium from 112 MDS patients and correlated them with French-American British (FAB) type, the international prognostic scoring system (IPSS), karyotype, peripheral blood (PB) and bone marrow (BM) blast cells, cytopenias, lactate dehydrogenase (LDH), and survival data. Concerning the FAB classification, lower median mCFUs were found in patients with refractory anemia (RA) and refractory anemia with ringed sideroblasts (RARS) compared to refractory anemia with excess of blast cells (RAEB) and refractory anemia with excess of blasts cells in transformation (RAEB-T). In vitro growth in MDS clearly correlated with the cytogenetic risk groups defined by the IPSS (30.5/10(5) PBMCs with favorable karyotypes, 191 in the intermediate prognostic group, 677 with unfavorable cytogenetics, p=0.015 favorable vs unfavorable). BM blast cells >5% (60.5 vs 255 colonies, p=0.032) as well as LDH levels above the normal limit (64.5 vs 425 colonies, p=0.045) were also associated with higher colony formation. Patients were stratified according to the number of circulating mCFUs into a low growth, intermediate growth and high growth group. Median survival was 343 days in the high growth, 1119 days in the low growth, and 2341 days in the intermediate growth group ( p=0.0002). Multivariate analyses revealed colony growth ( p=0.0056), PB blast cells ( p=0.0069), cytogenetic risk group ( p=0.024), and platelet count ( p=0.018) to predict survival in our patients. After inclusion of the IPSS risk categories, mCFU levels remained a highly predictive parameter for survival ( p=0.0056) and acute myeloblastic leukemia (AML) transformation ( p=0.0003). PMID- 12739064 TI - Efficacy and safety of splenectomy in adult chronic immune thrombocytopenia. AB - For patients with adult chronic immune thrombocytopenia (ITP) splenectomy (SE) is a highly effective treatment, but there are still uncertainties regarding the long-term efficacy and safety. We evaluated the long-term efficacy and safety of SE in 48 consecutive adult patients with chronic ITP (26 women, 22 men) who underwent SE between 1990 and 2001 at the General Hospital in Vienna, Austria. All patients had no remission after steroid treatment and were steroid dependent. The median age at the time of SE was 44 years (range: 16-77 years). Of 48 patients, 37 achieved a complete remission (CR, platelet count >100 x 10(9)/l), 8 a partial remission (PR) (platelet count 30-100 x 10(9)/l), and 2 had no response (NR). The probability of the overall survival was 98% at a median postsplenectomy observation time of 3.5 years. Seven patients with CR and four patients with PR relapsed. There were no relapses after 1 year. The probability of continuous complete remission (CCR) at 10 years was 79%. The probability of having a platelet count of >100 x 10(9)/l or >30 x 10(9)/l was 61% and 67%, respectively, at 5 and 10 years after splenectomy. Of the 11 relapsed patients, 5 had a second CR ( n=3) or PR ( n=2). The postoperative platelet count was the best predictor for a long-term remission. All patients with postoperative platelet counts >250 x 10(9)/l remained in CR. Patients aged >45 years had a similar success rate as compared with younger patients. Three patients had infections (one pneumonia and two fever of unknown origin) requiring hospitalization, but none had overwhelming septicemia. PMID- 12739065 TI - Leukemoid reaction response to chemotherapy and radiotherapy in a patient with cervical carcinoma. AB - A white blood cell count more than 50 x 10(9)/l associated with a cause outside the bone marrow is termed a leukemoid reaction. Although it simulates leukemia, most of its causes are benign. Malignancy as a cause of a leukemoid reaction is still a medical dilemma. It is thought to be attributed to granulocyte colony stimulating factor (G-CSF) secreted by the tumor cells. To our knowledge this is the first time a leukemoid reaction has been reported in association with cervical cancer. We even managed to monitor the leukemoid reaction response to chemotherapy and radiotherapy. PMID- 12739067 TI - Optimization of dendritic cell maturation and gene transfer by recombinant adenovirus. AB - Dendritic cells (DC) have vast potential for immunotherapy. Transferring therapeutic genes to DC may enhance their inherent T cell-stimulatory capacity. Recombinant adenovirus is the most efficient vehicle for DC gene transfer and can alone mature DC. We sought to define the parameters of adenovirus infection of murine bone marrow-derived DC (BMDC) and the concomitant impact on BMDC maturation. The efficiency of adenoviral gene transfer to DC depended on the mouse strain, the organ source of DC, and the level of DC maturation. C57BL/6 BMDC consistently had higher transgene expression than BALB/c DC. While BMDC had considerable GFP expression after AdGFP infection, adenovirus was relatively ineffective in accomplishing transgene expression in freshly isolated hepatic or splenic DC. BMDC that were relatively immature because of a shorter duration of culture had higher transgene expression after infection. Nevertheless, pretreatment of DC with exogenous stimulants such as LPS or TNF-alpha resulted in higher transgene expression. Maturation of BMDC depended only on virus entry but not viral gene or transgene expression. Therefore, DC maturation was disproportionately high compared to the percentage of DC that actually expressed the adenoviral transgene. Maturation by adenovirus was only seen in BMDC, but not in liver or splenic DC, and was more pronounced in DC from later in culture (day 12 versus day 6). There was a dose-response relationship, up to a threshold dose, between adenovirus infection and both DC maturation and enhancement of DC activation of antigen-specific T cells. Our findings underscore the importance of optimizing gene transfer to DC in designing strategies for immunotherapy. PMID- 12739068 TI - Interferon-alpha and interleukin-12 gene therapy of cancer: interferon-alpha induces tumor-specific immune responses while interleukin-12 stimulates non specific killing. AB - Cytokine gene therapy is applied in clinical studies of tumors, and IFN-alpha and IL-12 are widely used for cancer immunotherapy. Using a poorly immunogenic murine colorectal cancer cell line, MC38, we compared antitumor effects of IFN-alpha and IL-12. Transduced MC38 cell lines expressing IFN-alpha or IL-12 (MC38-IFNalpha or MC38-IL12, respectively) were established using retroviral vectors. Transduction of IFN-alpha or IL-12 gene to MC38 cells significantly reduced tumorigenicity in immunocompetent mice. When tumor-free mice initially injected with MC38-IFNalpha or MC38-IL12 cells were reinjected contralaterally with wild-type MC38 cells (MC38-WT) after 35 days, 7 of 12 or 2 of 12 mice rejected MC38-WT cells, respectively. In therapy-model mice with established tumor derived from MC38-WT cells, inoculation of gene-transduced cells significantly suppressed growth of the tumor in MC38-IFNalpha-inoculated groups, but not in the IL-12-inoculated group. Immunohistologic and flow cytometric analyses showed marked infiltration of CD8(+) cells in wild-type tumors of mice inoculated with IFN-alpha-expressing cells. Leukocyte-depletion experiments implicated CD8(+) T cells in tumor rejection induced by IFN-alpha-transduction; both CD8(+) T cells and natural killer cells were implicated in the more modest antitumor effect from IL-12 expression. To investigate induction of tumor-specific immune responses, we stimulated splenocytes from tumor-free mice twice in vitro with genetically modified MC38 cells. In vitro stimulations with MC38-IFNalpha cells induced definite MC38-specific lysis, but not stimulations with MC38-IL-12 cells. Injecting combination of MC38-IFNalpha and MC38-IL-12 cells caused an additive antitumor effect in the therapy model. These data suggested that IFN-alpha induces cytotoxic T lymphocytes and elicits long-lasting tumor-specific immunity, whereas IL-12 seems to stimulate non-specific killing. With additional refinements, combined IFN-alpha and IL-12 gene therapy might warrant clinical trials. PMID- 12739069 TI - High levels of Fas ligand and MHC class II in the absence of CD80 or CD86 expression and a decreased CD4+ T cell Infiltration, enables murine skin tumours to progress. AB - It is still not clear why some tumours will be recognized and destroyed by the immune system, and others will persist, grow, and eventually kill the host. It has been hypothesized that tumour cells might evade immunological destruction by expressing Fas ligand (FasL), a molecule which induces apoptosis in Fas(+) target cells. However, the role of FasL in creating an immune privileged status within a tumour remains controversial. To determine whether FasL is associated with skin tumour progression, we developed a tumour model enabling us to compare two squamous cell carcinomas (SCC). One is a regressor SCC which spontaneously regresses after injection into syngeneic mice. The other is a progressor SCC which evades immunological destruction. Detailed flow cytometric analysis was used to study tumour cell expression of FasL, Fas, CD80, CD86 and MHC class II. We also analysed the percentage of apoptotic tumour cells in vivo using annexin V and correlated skin tumour progression with CD4 and CD8 T cell infiltration. Progressor tumours expressed high levels of FasL in vivo, which was virtually absent from regressor tumours. The percentage of progressor tumours expressing MHC II was significantly greater than regressor tumours, while neither tumour expressed CD80 or CD86 costimulatory molecules. Consistent with a regressor phenotype, the percentage of viable tumour cells was significantly lower for regressor compared to progressor tumours which coincided with a significantly larger CD4(+) T cell infiltrate into the tumour mass. The results suggest that progression of skin tumours occurs if tumour cells express high levels of MHC II but not costimulatory molecules such as CD80 or CD86. This implies that tumours may induce anergy in CD4(+) T cells via MHC II antigen presentation in the absence of costimulation. To ensure escape from the immune system, tumours may then kill these T cells via a FasL-dependent mechanism. PMID- 12739070 TI - [18F]FDG imaging of head and neck tumours: comparison of hybrid PET and morphological methods. AB - The aim of this study was to evaluate fluorine-18 fluorodeoxyglucose ([(18)F]FDG) imaging of head and neck tumours using a second- or third-generation hybrid PET device. Results were compared with the findings of spiral computed tomography (CT) and magnetic resonance imaging (MRI), and, as regards lymph node metastasis, the ultrasound findings. A total of 116 patients with head and neck tumours (83 males and 33 females aged 27-88 years) were examined using a hybrid PET scanner after injection of 185-350 MBq of [(18)F]FDG (Picker Prism 2000 XP-PCD, Marconi Axis gamma-PET(2) AZ). Hybrid PET examinations were performed in list mode using an axial filter. Reconstruction of data was performed iteratively. Ninety-six patients underwent CT using a multislice technique (Siemens Somatom Plus 4, Marconi MX 8000), 18 patients underwent MRI and 100 patients were examined by ultrasound. All findings were verified by histology, which was considered the gold standard, or, in the event of negative histology, by follow-up. Correct diagnosis of the primary or recurrent lesion was made in 73 of 85 patients using the hybrid PET scanner, in 50 of 76 patients on CT and in 7 of 10 patients on MRI. Hybrid PET successfully visualised metastatic disease in cervical lymph nodes in 28 of 34 patients, while 23 of 31 were correctly diagnosed with CT, 3 of 4 with MRI and 30 of 33 with ultrasound. False positive results regarding lymph node metastasis were seen in three patients with hybrid PET, in 14 patients with CT and in 13 patients with ultrasound. MRI yielded no false positive results concerning lymph node metastasis. In one patient, unrecognised metastatic lesions were seen on hybrid PET elsewhere in the body (lung: n=1; bone: n=1). Additional malignant lesions at sites other than the head and neck tumour were found in three patients (one patient with lung cancer, one patient with pelvic metastasis due to a carcinoma of the prostate and one patient with pulmonary metastasis due to breast cancer). It is concluded that [(18)F]FDG PET with hybrid PET scanners is superior to CT and MRI in the diagnosis of primary or recurrent lesions as well as in the assessment of lymph node involvement, whereas it is inferior to ultrasound in the detection of cervical lymph node metastasis. PMID- 12739071 TI - Potential impact of [18F]3'-deoxy-3'-fluorothymidine versus [18F]fluoro-2-deoxy-D glucose in positron emission tomography for colorectal cancer. AB - Fluorine-18 labelled fluoro-2-deoxy- d-glucose ((18)FDG) positron emission tomography (PET) imaging demonstrates the increased glucose consumption of malignant cells, but problems with specificity have led to the development of new PET tracers. [(18)F]3'-deoxy-3'-fluorothymidine ((18)FLT) is a new tracer which images cellular proliferation by entering the salvage pathway of DNA synthesis. In this study we compared the cellular uptake of (18)FLT and (18)FDG in patients with colorectal cancer (CRC). Seventeen patients with 50 primary or metastatic CRC lesions were prospectively recruited. Lesions were initially identified using computed tomography. Patients underwent both (18)FDG and (18)FLT scanning. Semi quantitative analysis of tracer uptake was carried out using standardised uptake values. All the primary tumours ( n=6) were visualised by both tracers, with (18)FDG showing on average twice the uptake of (18)FLT. Similar uptake of both tracers was seen in lung and peritoneal lesions, with (18)FLT imaging five of the six lung lesions and all of the peritoneal lesions. Of the 32 colorectal liver metastases, 11 (34%) were seen as avid for (18)FLT, compared with 31 (97%) for (18)FDG. No correlation was seen between the uptake of the two tracers ( R(2)=0.03). (18)FLT shows a high sensitivity in the detection of extrahepatic disease but poor sensitivity for the imaging of colorectal liver metastases, making it unlikely to have a role as a diagnostic tracer in CRC. We have demonstrated that (18)FDG and (18)FLT image two distinct processes. The prognostic implications of the uptake of (18)FLT need to be assessed in terms of response to chemoradiotherapy and survival. PMID- 12739072 TI - Analysis of actin dynamics at the leading edge of crawling cells: implications for the shape of keratocyte lamellipodia. AB - Leading edge protrusion is one of the critical events in the cell motility cycle and it is believed to be driven by the assembly of the actin network. The concept of dendritic nucleation of actin filaments provides a basis for understanding the organization and dynamics of the actin network at the molecular level. At a larger scale, the dynamic geometry of the cell edge has been described in terms of the graded radial extension model, but this level of description has not yet been linked to the molecular dynamics. Here, we measure the graded distribution of actin filament density along the leading edge of fish epidermal keratocytes. We develop a mathematical model relating dendritic nucleation to the long-range actin distribution and the shape of the leading edge. In this model, a steady state graded actin distribution evolves as a result of branching, growth and capping of actin filaments in a finite area of the leading edge. We model the shape of the leading edge as a product of the extension of the actin network, which depends on actin filament density. The feedback between the actin density and edge shape in the model results in a cell shape and an actin distribution similar to those experimentally observed. Thus, we explain the stability of the keratocyte shape in terms of the self-organization of the branching actin network. PMID- 12739073 TI - Transformation of Azospirillum brasilense Cd with an ACC deaminase gene from enterobacter cloacae UW4 fused to the Tet r gene promoter improves its fitness and plant growth promoting ability. AB - It has been reported that PGPB, containing ACC deaminase, can cleave the plant ethylene precursor ACC and thereby lower ethylene concentration in a developing or stressed plant, protecting it against the deleterious effects of stress ethylene and facilitating the formation of longer roots. In a previous work we have demonstrated expression of the ACC deaminase gene ( acdS) from Enterobacter cloacae UW4 under the control of the lac promoter in Azospirillum brasilense Cd. With the inference that a construct including the ACC deaminase gene under the control of a constitutive promoter weaker than the lac promoter might impose less metabolic load on Azospirillum and improve its fitness, it was decided to clone acdS under the control of a tetracycline resistance gene promoter. The ACC deaminase structural gene was fused to the Tet(r) gene promoter by overlap extension using PCR, cloned in pRK415, and transferred into A. brasilense Cd. The resulting transformants showed lower ACC deaminase activity than those with the lac promoter controlled acdS gene. However, acdS under the control of the Tet(r) gene promoter imposed lesser metabolic load on Azospirillum brasilense Cd. The result was significantly increased IAA synthesis and greater bacterial growth rate, as well as increased ability to survive on the surface of tomato leaves and to promote the growth of tomato seedlings. PMID- 12739074 TI - Bacterial diversity in a marine methanol-fed denitrification reactor at the montreal biodome, Canada. AB - The bacterial biota of a methanol-fed denitrification reactor used to treat seawater at the Montreal Biodome were investigated using culture-dependent and molecular biology methods. The microbiota extracted from the reactor carriers were cultivated on three media. Three isolate types were recovered and their 16S ribosomal DNA (rDNA) genes were determined. The analysis showed that the isolate types were related to alpha-Proteobacteria. They are members of the Hyphomicrobium and Paracoccus genera and the Phyllobacteriaceae family. Uncultured bacteria were identified through a 16S rDNA library generated from total DNA extracted from the microbiota. Clones were screened for different restriction profiles and for different DGGE (denaturing gradient gel electrophoresis) migration profiles. More than 70% of clones have the same restriction profile, and the sequence of representative clones showed a relation with the Methylophaga members of the Piscirickettsia family (gamma Proteobacteria). Sequences from other profiles were related to bacterial species involved in denitrification. The number of species in the denitrification reactor was estimated at 15. Bacterial colonization on newly added carriers in the denitrification reactor was monitored by PCR-DGGE. The DGGE migration profiles evolved during the first 5 weeks and then remained essentially unchanged. PCR DGGE was also used to monitor the microbial profiles in various aquarium locations. As expected, bacterial populations differed from one location to another, except for the sand and trickling filters which presented similar DGGE migration profiles. PMID- 12739075 TI - Simultaneous measurements of organic carbon mineralization and bacterial production in oxic and anoxic lake sediments. AB - Based on work in marine sediments it can be hypothesized that (i) overall OM mineralization depends on the enzymatic capacity and is largely independent from the energy yield, (ii) similar oxic and anoxic rates are expected for fresh OM, while oxic rates should be faster for old OM that is partially degraded or adsorbed to particles, and (iii) that the thermodynamic energy yield does not regulate mineralization, but primarily determines the energy fraction allocated to bacterial production (BP). We addressed these hypotheses by simultaneous measurements of mineralization rates (MR) and BP in sediments from a eutrophic lake, along with MR measurements in sediments of a dystrophic lake. Anoxic MR were 44 and 78% of oxic MR in the eutrophic and dystrophic lake, respectively, which was always higher than expected given the theoretical energy yields. The BP:MR ratio was 0.94 and 0.24 in the oxic and anoxic treatments, respectively, in accordance with the expected energy yields. Thus, the results support all three hypotheses above. We also critically discuss BP measurements in sediments and suggest that bacterial growth efficiency values from simultaneous MR and BP measurements can be used to evaluate the reliability of BP estimates. PMID- 12739076 TI - Root-parasitic nematodes enhance soil microbial activities and nitrogen mineralization. AB - Obligate root-parasitic nematodes can affect soil microbes positively by enhancing C and nutrient leakage from roots but negatively by restricting total root growth. However, it is unclear how the resulting changes in C availability affect soil microbial activities and N cycling. In a microplot experiment, effects of root-parasitic reniform nematodes ( Rotylenchulus reniformis) on soil microbial biomass and activities were examined in six different soils planted with cotton. Rotylenchulus reniformis was introduced at 900 nematodes kg(-1) soil in May 2000 prior to seeding cotton. In 2001, soil samples were collected in May before cotton was seeded and in November at the final harvest. Extractable C and N were consistently higher in the R. reniformis treatments than in the non nematode controls across the six different soils. Nematode inoculation significantly reduced microbial biomass C, but increased microbial biomass N, leading to marked decreases in microbial biomass C:N ratios. Soil microbial respiration and net N mineralization rates were also consistently higher in the nematode treatments than in the controls. However, soil types did not have a significant impact on the effects of nematodes on these microbial parameters. These findings indicate that nematode infection of plant roots may enhance microbial activities and the turnover of soil microbial biomass, facilitating soil N cycling. The present study provides the first evidence about the direct role of root-feeding nematodes in enhancing soil N mineralization. PMID- 12739077 TI - Sediment microbes of deep-sea bioherms on the northwest shelf of Australia. AB - The northwest shelf of Australia is a region with known petroleum reserves. Recent geological investigations have revealed carbonate knolls postulated to be of biological origin. Sediment microbial populations at three sites on the northwest shelf, Pee Shoal, Mermaid Reef, and Scott Reef, were investigated for the presence of hydrocarbon-degrading bacteria. From two sampling trips, 246 bacterial strains were purified, and 182 of these strains displayed preferential growth on hydrocarbon-selective media. Strains were tested for nutrient specificity using single hydrocarbon fermentations. Metabolic profiles were constructed using biochemical testing, fatty acid analysis, and metabolic rate experiments. 16S rRNA gene sequence analysis of four strains identified these strains as members of the genera Sphingomonas, Bacillus, and Microbacterium, and one strain as a member of the alpha-Proteobacteria. The potential of these strains as bioindicators of hydrocarbon pollutants or for bioremediation in marine environments is discussed. PMID- 12739078 TI - Bacterioplankton community structure in a maritime antarctic oligotrophic lake during a period of holomixis, as determined by denaturing gradient gel electrophoresis (DGGE) and fluorescence in situ hybridization (FISH). AB - The bacterioplankton community structure in Moss Lake, a maritime Antarctic oligotrophic lake, was determined with vertical depth in the water column, during the ice-free period on Signy Island in the South Orkney Islands. Bacterioplankton community structure was determined using a combination of direct counting of 4',6 diamidino-2-phenylindole (DAPI) stained cells, PCR amplification of 16S rRNA gene fragments, denaturing gradient gel electrophoresis (DGGE) and in situ hybridization with group-specific, fluorescently labeled oligonucleotide probes. Using PCR amplification of 16S rRNA gene fragments and DGGE, the bacterioplankton community composition was shown to be constant with vertical depth in the water column. Specific bacterioplankton species identified through cloning and sequencing the DGGE products obtained were Flavobacterium xinjiangensis (a Flavobacterium), Leptothrix discophora (a beta-Proteobacterium), and a number of uncultured groups: two beta-Proteobacteria, an unclassified Proteobacterium, three sequences from Actinobacteria, and a Cyanobacterium. Fluorescence in situ hybridization (FISH), however, demonstrated that there were minor but significant fluctuations in different groups of bacteria with vertical depth in the water column. It showed that the beta-Proteobacteria accounted for between 26.4 and 71.5%, the alpha-Proteobacteria 2.3-10.6%, the gamma-Proteobacteria 0-29.6%, and the Cytophaga-Flavobacterium group 1.8-23.5% of cells hybridizing to a universal probe. This study reports the first description of the community structure of an oligotrophic Antarctic freshwater lake as determined by PCR-dependent and PCR independent molecular techniques. It also suggests that the bacterioplankton community of Moss Lake contains classes of bacteria known to be important in freshwater systems elsewhere in the world. PMID- 12739079 TI - Benthic bacterial production and protozoan predation in a silty freshwater environment. AB - The interrelation of heterotrophic bacteria with bacterivorous protists has been widely studied in pelagic environments, but data on benthic habitats, especially in freshwater systems, are still scarce. We present a seasonal study focusing on bacterivory by heterotrophic nanoflagellates (HNF) and ciliates in the silty sediment of a temperate macrophyte-dominated oxbow lake. From January 2001 to February 2002 we monitored the standing stock of bacteria and protozoa, bacterial secondary production (BSP, (3)H-thymidine, and (14)C-leucine incorporation), and grazing rates of HNF and ciliates on bacteria (FLB uptake) in the oxic sediment of the investigated system. BSP ranged from 470 to 4050 micro g C L(-1) wet sediment h(-1). The bacterial compartment turned out to be highly dynamic, indicated by population doubling times (0.6-10.0 d), which were comparable to those in the water column of the investigated system. Yet, the control mechanisms acting upon the bacterial population led to a relative constancy of bacterial standing stock during a year. Ingestion rates of protozoan grazers were 0-20.0 bacteria HNF(-1) h(-1) and 0-97.6 bacteria ciliate(-1) h(-1). HNF and ciliates together cropped 0-14 (mean 4)% of BSP, indicating that they did not significantly contribute to benthic bacterial mortality during any period of the year. The low impact of protozoan grazing was due to the low numbers of HNF and ciliates in relation to bacteria (1.8-3.5 x 10(4) bacteria HNF(-1), 0.9-3.1 x 10(6) bacteria ciliate(-1)). Thus, grazing by HNF and ciliates could be ruled out as a parameter regulating bacterial standing stock or production in the sediment of the investigated system, but the factors responsible for the limitation of benthic protistan densities and the fate of benthic BSP remained unclear. PMID- 12739080 TI - Factors controlling extremely productive heterotrophic bacterial communities in shallow soda pools. AB - Dilute soda lakes are among the world's most productive environments and are usually dominated by dense blooms of cyanobacteria. Up to now, there has been little information available on heterotrophic bacterial abundance, production, and their controlling factors in these ecosystems. In the present study the main environmental factors responsible for the control of the heterotrophic bacterial community in five shallow soda pools in Eastern Austria were investigated during an annual cycle. Extremely high cyanobacterial numbers and heterotrophic bacterial numbers up to 307 x 10(9) L(-1) and 268 x 10(9) L(-1) were found, respectively. Bacterial secondary production rates up to 738 micro g C L(-1) h( 1) and specific growth rates up to 1.65 h(-1) were recorded in summer and represent the highest reported values for natural aquatic ecosystems. The combination of dense phytoplankton blooms, high temperature, high turbidity, and nutrient concentration due to evaporation is supposed to enable the development of such extremely productive microbial populations. By principal component analysis containing the data set of all five investigated pools, two factors were extracted which explained 62.5% of the total variation of the systems. The first factor could be interpreted as a turbidity factor; the second was assigned to as concentration factor. From this it was deduced that bacterial and cyanobacterial abundance were mainly controlled by wind-induced sediment resuspension and turbidity stabilized by the high pH and salinity and less by evaporative concentration of salinity and dissolved organic carbon. Bacterial production was clustered with temperature in factor 3, showing that bacterial growth was mainly controlled by temperature. The concept of describing the turbid water columns of the shallow soda pools as "fluid sediment" is discussed. PMID- 12739081 TI - Genetic characterization of the nitrate reducing community based on narG nucleotide sequence analysis. AB - The ability of facultative anerobes to respire nitrate has been ascribed mainly to the activity of a membrane-bound nitrate reductase encoded by the narGHJI operon. Respiratory nitrate reduction is the first step of the denitrification pathway, which is considered as an important soil process since it contributes to the global cycling of nitrogen. In this study, we employed direct PCR, cloning, and sequencing of narG gene fragments to determine the diversity of nitrate reducing bacteria occurring in soil and in the maize rhizosphere. Libraries containing 727 clones in total were screened by restriction fragment analysis. Phylogenetic analysis of 128 narG sequences separated the clone families into two main groups that represent the Gram-positive and Gram-negative nitrate-reducing bacteria. Novel narG lineages that branch distinctly from all currently known membrane bound nitrate-reductase encoding genes were detected within the Gram negative branch. All together, our results revealed a more complex nitrate reducing community than did previous culture-based studies. A significant and consistent shift in the relative abundance of the nitrate-reducing groups within this functional community was detected in the maize rhizosphere. Thus a substantially higher abundance of the dominant clone family and a lower diversity index were observed in the rhizosphere compared to the unplanted soil, suggesting that a bacterial group has been specifically selected within the nitrate-reducing community. Furthermore, restriction fragment length polymorphism analysis of cloned narG gene fragments proved to be a powerful tool in evaluating the structure and the diversity of the nitrate-reducing community and community shifts therein. PMID- 12739082 TI - The radiological spectrum of invasive aspergillosis in children: a 10-year review. AB - BACKGROUND: Invasive aspergillosis is an uncommon but life-threatening event in the immunocompromised child. Attempts at fungal isolation are often unrewarding and a high index of radiological suspicion is essential in the early diagnosis of infected children. OBJECTIVE: To document the radiological spectrum of disease in invasive aspergillosis in the paediatric population. MATERIALS AND METHODS: A retrospective review of the imaging performed in 27 consecutive patients (age 7 months to 18 years) with documented invasive Aspergillosis encountered over a 10 year period at a single institution. RESULTS: Radiographic findings of pulmonary disease (20 patients) included segmental and multilobar consolidation, perihilar infiltrates, multiple small nodules, peripheral nodular masses and pleural effusions. No cavitating lesions were seen on CXR. Small cavitating nodules were present on CT in two of eight children. Chest wall disease was particularly associated with underlying chronic granulomatous disease. Disseminated disease manifested as osteomyelitis (n=5), cerebral (n=3), oesophageal (n=1), hepatic (n=2), renal (n=2) and cutaneous (n=5) involvement. Imaging findings are discussed. Twelve patients (44%) subsequently died from Aspergillus-related complications. CONCLUSIONS: Invasive aspergillosis presents with a wide variety of radiographic findings involving multiple organ systems. Respiratory findings are varied but often non-specific, and a high index of suspicion is necessary in immunocompromised patients. In contrast to adult disease, the incidence of cavitation of pulmonary lesions appears low. PMID- 12739083 TI - Influence of movement speed on accuracy and coordination of reaching movements to memorized targets in three-dimensional space in a deafferented subject. AB - Multiarticular reaching movements at different speeds produce differential demands for the on-line control of ongoing movements and for the predictive control of intersegmental dynamics. The aim of this study was to assess the ability of a proprioceptively deafferented patient and aged-matched control subjects to make precise and coordinated three-dimensional reaching movements at different speeds without vision during the movement. A patient with a complete loss of proprioception below the neck (C.F.) and five control subjects made reaching movements to four remembered visual targets at slow, natural, and fast speeds. All movements were performed without vision of the arm during the movements. The spatial accuracy, the movement kinematics and the interjoint coordination of these movements were analyzed. Results showed that control subjects made larger spatial errors at both slow and fast speeds than at natural speed. However, they synchronized motions at the shoulder and elbow joints and kept most movement kinematic features invariant across speed conditions. In contrast, C.F. failed to produce smooth and simultaneous motions at the shoulder and elbow joints at all speeds. Surprisingly, however, he made much larger errors than control subjects at slow and natural speeds, but not at fast speed. Analysis of patterns of interjoint coordination revealed that, when instructed to move fast, C.F. initiated arm movements by fixing the elbow while moving the shoulder joint to damp interaction torques exerted on the elbow joint from motion of the upper arm. The results demonstrated that, although proprioceptive loss disrupted normal control of multijoint movements at all speeds, when performing relatively fast three-dimensional movements, C.F. could control intersegmental dynamics by reducing the number of active joints. More importantly, the results highlight the dual role of proprioception in controlling multijoint movements; that is, to provide important cues both for the predictive control of interaction torques and for the synchronization of adjacent joints even when interactive torques are very small. These findings support the idea that proprioceptive input is used by the CNS to update an internal model of limb dynamics that adapts the motor plan according to biomechanical contexts. PMID- 12739084 TI - Frequency-dependent effects of muscle tendon vibration on corticospinal excitability: a TMS study. AB - The aim of the present study was to investigate the effects of muscle tendon vibration at different frequencies on corticospinal excitability by means of transcranial magnetic stimulation (TMS). A second objective was to describe whether the observed modulations in motor evoked potentials (MEPs), as a function of vibration frequency, reflect the behavior of Ia afferents during and after vibration. In ten subjects, muscle tendon vibration (duration 30 s) was applied to the flexor carpi radialis (FCR) muscle at three different frequencies (20, 75 and 120 Hz). MEPs following single-pulse TMS were recorded from the targeted muscle during a previbration, vibration, and postvibration period. Muscle tendon vibration at 75 Hz increased the MEP amplitude significantly during vibration, whereas a smaller but still significant effect was observed during 120 Hz vibration. No significant MEP changes could be observed during 20 Hz vibration and during the postvibration period for each frequency. Our findings indicate that muscle tendon vibration exerts a frequency-dependent effect on corticospinal excitability. Furthermore, evidence is provided for the notion that the excitatory effect of muscle tendon vibration on the primary motor cortex is mediated by Ia afferent input. PMID- 12739086 TI - Differential modulation of tremor and pulsatile control of human jaw and finger by experimental muscle pain. AB - Resting tremor is seen in both the limbs and in the trigeminal motor system. These rhythmical perturbations are the result of alternating activation of antagonistic muscles, and these increase in amplitude during slow, voluntary movements. In the present study, we examined the effect of experimental muscle pain on finger and jaw tremor. The tremor in the mandible and in the middle finger was measured on separate occasions, at rest and during two constant velocity movements. Pain was then induced by the infusion of hypertonic saline into a jaw-closing muscle (masseter) or into a finger extensor muscle (extensor digitorum longus, EDL). During masseter pain, the power at the peak tremor frequency of the mandible decreased significantly both when the jaw was at rest and during voluntary jaw movements at two velocities. In contrast, pain in EDL resulted in a significant increase in the power of finger tremor only during the two speeds of voluntary movement. No change in the peak tremor frequency was seen in either the finger or the jaw during pain. The most likely explanation for these data is that muscle pain tonically modulates the amplitude of the outputs from the central "pulsatile control" generators that drive the alternating activation of antagonistic muscles which produce tremor at rest and during movements. This modulation is in the opposite direction for systems controlling jaw and hand, suggesting a specific interaction of the nociceptive afferents with separate central oscillators. PMID- 12739085 TI - Evidence for a unimodal somatosensory attention system. AB - Extinction is generally viewed as a disorder of selective attention for spatial locations. Recent physiologic, behavioral and lesion studies view spatial locations as a complex construct in which multiple modalities and motor systems are integrated. Accordingly, cross-modal and sensory-motor conditions often modify extinction. In a patient with tactile extinction, we tested the hypothesis that attentional deficits can also be confined to a specific sensory modality. Using objectively and subjectively balanced tactile stimuli and signal detection analysis, we found that our patient's contralesional tactile discrimination was not modulated by proprioceptive or visual input or by movement. By contrast, increasing the salience of the contralesional tactile stimuli did improve her contralesional tactile discrimination, consistent with our hypothesis that she suffered from a modality-specific attentional deficit. Additionally, she did not have any evidence of visual extinction, again bolstering our claim that her extinction was confined to touch. These data suggest that in addition to polymodal and sensory-motor attentional systems, spatial attention also operates on specific sensations. We also advocate the use of signal detection analysis, a method that has been surprisingly neglected in extinction research. PMID- 12739087 TI - Postural control in children. Coupling to dynamic somatosensory information. AB - The purpose of this investigation was to determine whether the coupling between dynamic somatosensory information and body sway is similar in children and adults. Thirty children (4-, 6-, and 8-year-olds) and 10 adults stood upright, with feet parallel, and lightly contacting the fingertip to a rigid metal plate that moved rhythmically at 0.2, 0.5, and 0.8 Hz. Light touch to the moving contact surface induced postural sway in all participants. The somatosensory stimulus produced a broadband frequency response in children, while the adult response was primarily at the driving frequency. Gain, as a function of frequency, was qualitatively the same in children and adults. Phase decreased less in 4-year-olds than other age groups, suggesting a weaker coupling to position information in the sensory stimulus. Postural sway variability was larger in children than adults. These findings suggest that, even as young as age 6, children show well-developed coupling to the sensory stimulus. However, unlike adults, this coupling is not well focused at the frequency specified by the somatosensory signal. Children may be unable to uncouple from sensory information that is less relevant to the task, resulting in a broadband response in their frequency spectrum. Moreover, higher sway variability may not result from the sensory feedback process, but rather from the children's underdeveloped ability to estimate an internal model of body orientation. PMID- 12739088 TI - Ia Afferent input alters the recruitment thresholds and firing rates of single human motor units. AB - Vibration of the patellar tendon recruits motor units in the knee extensors via excitation of muscle spindles and subsequent Ia afferent input to the alpha motoneuron pool. Our first purpose was to determine if the recruitment threshold and firing rate of the same motor unit differed when recruited involuntarily via reflex or voluntarily via descending spinal pathways. Although Ia input is excitatory to the alpha-motoneuron pool, it has also been shown paradoxically to inhibit itself. Our second purpose was to determine if vibration of the patellar tendon during a voluntary knee extension causes a change in the firing rate of already recruited motor units. In the first protocol, 10 subjects voluntarily reproduced the same isometric force profile of the knee extensors that was elicited by vibration of the patellar tendon. Single motor unit recordings from the vastus lateralis (VL) were obtained with tungsten microelectrodes and unitary behaviour was examined during both reflex and voluntary knee extensions. Recordings from 135 single motor units showed that both recruitment thresholds and firing rates were lower during reflex contractions. In the second protocol, 7 subjects maintained a voluntary knee extension at 30 N for approximately 40-45 s. Three bursts of patellar tendon vibration were superimposed at regular intervals throughout the contraction and changes in the firing rate of already recruited motor units were examined. A total of 35 motor units were recorded and each burst of superimposed vibration caused a momentary reduction in the firing rates and recruitment of additional units. Our data provide evidence that Ia input modulates the recruitment thresholds and firing rates of motor units providing more flexibility within the neuromuscular system to grade force at low levels of force production. PMID- 12739089 TI - Attenuation of human neck muscle activity following repeated imposed trunk forward linear acceleration. AB - It has been suggested that, after a passive linear acceleration of a seated subject which resembles a small, rear-end car impact, sensory information from proprioceptive, vestibular, and visual systems elicit stabilizing neck muscular responses. These neck muscular responses are presumably reflex based and are modified with the magnitude of the perturbation. A key issue that remains is to determine whether the neck and head postural responses can be modulated by a previous experience of the acceleration and not only by the magnitude of the acceleration. This question is of interest because, contrary to cadaver studies, one could expect that humans apprehending a rapid trunk acceleration would adopt a bracing behavior to minimize head movements. The aim of the present experiment was to verify whether neck-muscle activities can be modulated when prior knowledge about whole-body acceleration onset, direction, and magnitude are unknown compared with when only acceleration onset is unknown. Nine seated subjects were submitted to 11 imposed, forward linear accelerations (1.1 g). For the first trial, subjects were completely unaware of the platform acceleration characteristics (onset, direction, amplitude, and acceleration magnitude). For the subsequent ten trials, subjects knew they would be submitted to a forward linear acceleration, but the onset of the acceleration was unknown. Head kinematics and EMG responses of the neck muscles to the first perturbation were similar for all subjects (6.2 degrees head extension, EMG activity starting from 55 to 72 ms after platform onset). Following the first trial, however, all subjects showed a decreased neck EMG activity. Moreover, subjects responded in one of two ways across trials: one group of subjects ( n=5) maintained a constant head angular position and velocity, whereas the other group ( n=4) showed an increased head angular position (up to 12.6 degrees ) and velocity. This suggests that the first perturbation trial revealed a completely reactive response. After this initial trial, the responses observed may present a mixture of feedforward and feedback control. It is likely that whiplash injuries occur under conditions resembling those observed for the first trial only. If this is the case, the behavior for the following trials cannot be representative of injury mechanisms occurring in whiplash-like motion. Altogether, our results strongly suggest that, following repeated trunk linear accelerations of a constant magnitude, the nervous system prefers to minimize muscle stress instead of adopting a bracing strategy. PMID- 12739090 TI - Preservation of electrocortical brain activity during hypoxemia in preterm lambs. AB - Adequate cerebral perfusion is necessary to preserve cerebral O(2) supply in order to maintain brain cell function. Our aim was to assess the influence of gestational age on the response of cerebral hemodynamics to hypoxemia and to determine thresholds of cerebral O(2) supply for preservation of brain cell function in preterm born lambs. Lambs were delivered by hysterotomy at 141 (n=5), 134 (n=5) or 127 (n=7) days of gestation. Decreases in arterial oxygen content (CaO(2)) were induced by stepwise reduction of the fraction of O(2) in inspired air (FiO(2)). Mean arterial blood pressure (MABP), carotid artery blood flow (Qcar), and electrocortical brain activity as a measure of brain cell function, were continuously recorded. Cerebral arterial blood gases were analyzed at the end of each hypoxemic level to calculate CaO(2) and cerebral O(2) supply. In contrast to 141-day lambs, MABP could not be maintained in 134-day and 127-day lambs at levels of severe hypoxemia. Increases in Qcar were observed at levels of moderate hypoxemia in all gestational age-groups. Albeit Qcar increased further at levels of severe hypoxemia in the 141-day lambs, Qcar declined under these conditions in the 134-day and 127-day lambs. The threshold of cerebral O(2) supply for the preservation of brain cell function was however similar in all gestational age-groups (1.7 ml/min). It is concluded that the ability to maintain cerebral function during hypoxemia depends upon the ability to preserve cerebral O(2) supply by means of cerebral hemodynamic compensatory mechanisms, which are not fully matured until 96% of gestation. PMID- 12739091 TI - Discrete and cyclical units of action in a mixed target pair aiming task. AB - Two experiments addressed the issue of discrete and cyclical units as possible basic units of action that might be used to construct complex actions based on task constraints. The experiments examined the influence of low and high accuracy constraints on the end-effector's motion in rhythmical aiming movements. Both experiments utilized a Fitts-type task under three accuracy constraints: (1) big target pairing-low index of movement difficulty (ID), (2) small target pairing high ID, and (3) mixed target pairing-one target high ID and the other target low ID. Experiment I was a 1-degree-of-freedom ( df) task that required subjects to crossover the inside edge of targets in a target pair using elbow flexion extension motions. Experiment II used a 2- df task that required subjects to tap back and forth between targets in a target pair using a hand-held stylus. In both experiments, end-effector motion in the low ID condition was cyclical with the end-effector's motion consistent with a limit-cycle attractor description, while in the high ID condition end-effector motion was discrete and consistent with a fixed-point attractor description. The mixed target pairing produced both discrete and cyclical features in the end-effector's dynamics that suggested a functional linking of discrete and cyclical units of action as the optimal movement solution. Evidence supporting the above statements was found in the kinematic measures of movement time (MT), dwell time, proportion of MT accelerating and decelerating, and in a measure of harmonicity (Guiard 1993, Acta Psychol 82:139-159; Guiard 1997, Hum Mov Sci 16:97-131). Extended practice in the mixed target condition revealed a bias towards cyclical motion with practice. The results demonstrate that discrete and cyclical motion, represented as limit-cycle and fixed-point attractors, are basic units of action that the motor system uses in constructing more complex action sequences. The results are discussed with reference to coordinative structures and the generalized motor program as basic units of action. Issues pertaining to visual feedback processing and movement braking in rapid aiming tasks are also discussed. PMID- 12739092 TI - Delayed flanker effects on lateralized readiness potentials. AB - When participants were required to respond to a relevant central target and ignore irrelevant flanking stimuli, the flankers produced a response compatibility effect. Electrophysiological studies have shown that irrelevant flanker stimuli can affect the motor system. The present experiments further examined the characteristics of flanker effects on the motor system. Sixty participants responded in the flanker task to arrows (experiment 1) or letters (experiment 2). To examine time and extent of flanker effects on the motor system, target onset was delayed with blocked or random stimulus-onset asynchronies (SOA). With SOA of 0 and 100 ms, flanker effects on behavioral measures were reduced in random as compared to blocked conditions, but enhanced with SOA of 400 ms. With SOA of 400 ms, flanker effects on the early lateralized readiness potentials (LRP) were reduced in blocked as compared to random conditions, indicating that the onset of flanker effects on the LRP was delayed. Response-locked LRPs suggest that flanker and target stimuli activate the motor system successively. Findings challenge current theories of the flanker compatibility effect. PMID- 12739094 TI - Anodal vestibular stimulation does not suppress vestibular reflexes in human subjects. AB - Anodal current applied to the vestibular apparatus has previously been found to inhibit discharge from irregular vestibular afferents in squirrel monkeys. We wished to investigate whether anodal currents applied over the mastoid processes of human subjects would significantly reduce ongoing vestibular activity and thereby the size of galvanic-evoked vestibulospinal reflexes, measured by soleus electromyogram. Nine subjects were tested, of whom six subjects (five females, one male) provided data for the final analysis. Tonic anodal current was applied over one mastoid at 0 (baseline), 2, 4, 6 and 8 mA. The cathode was placed at C7. Superimposed on each intensity of tonic current were separate, short anodal currents of 4 mA, duration 20 ms, presented as 128 stimuli to the same side, and used to test vestibular responsiveness. These trials were then repeated with the anode overlying the contralateral mastoid. Short latency (SL) and medium latency (ML) reflexes were measured from the right soleus muscle. All six subjects used in the final analysis had readily identifiable reflexes to all stimuli. One-way ANOVA revealed no significant difference in the magnitude of the SL ( P=0.99) or ML ( P=0.96) components of the vestibulospinal reflexes across the group. Despite surface anodal currents of up to 8 mA, there was no consistent effect on the size of galvanic-evoked vestibulospinal reflexes. As 8 mA is close to the maximum intensity tolerated by volunteer subjects, our results indicate that anodal current applied over the mastoids is unlikely to be a useful means of suppressing vestibular function in human subjects. PMID- 12739093 TI - Brainstem projections of different branches of the vestibular nerve: an experimental study by transganglionic transport of horseradish peroxidase in the cat. III. The saccular nerve. AB - The brainstem projection of the saccular nerve was investigated by transganglionic tracing with horseradish peroxidase in the cat. The labeled fibers were located most caudally and laterally (dorsolaterally) in the vestibular nerve root. This was unique when compared with the locations of the fibers from the nerves of the other vestibular-end organs that were studied by the present authors by the same approach previously. In addition, such a comparison revealed a specific location, from lateral to medial, for the fibers from each of the five divisions of the vestibular nerve. In the present study, labeled fibers from the sacculus, of fine caliber, were found close to the restiform body, both medially and laterally, some even penetrating through this structure. Labeled terminals were present in cell group "y". This was unique, compared with the nerves from the other end organs. Such terminals were also found in the four main vestibular nuclei, except for the medial vestibular nucleus, where no labeled terminals could be detected. No labeled terminals were found in the interstitial nucleus of the vestibular nerve. Together with the findings from our previous studies, this suggests that, in contrast to the ampullar nerves, the nerves from the maculae do not project to this structure. This study confirms, but also extends, findings reported from a previous investigation in the cat, using an experimental degeneration technique. PMID- 12739095 TI - Liquid chromatography-fluorescence detection for simultaneous analysis of sulfonamide residues in honey. AB - A rapid, accurate LC analytical method has been developed for determination of eight sulfonamides (sulfacetamide, sulfapyridine, sulfamerazine, sulfamethoxypyridazine, sulfameter, sulfachloropyridazine, sulfamethoxazole and sulfadimethoxine) in honey. The sample was dissolved in phosphoric acid solution (pH 2). After filtration, the sample solution was cleaned by use of two solid phase extraction (SPE) cartridges-an aromatic sulfonic cation-exchange cartridge and an Oasis HLB cartridge. The eight sulfonamides were then derivatized with fluorescamine and the derivatives were determined by LC with fluorescence detection at excitation and emission wavelengths of 405 and 495 nm, respectively. Average recoveries at three fortification levels in the range 0.02-0.50 mg kg(-1) in twelve different kinds of honey were 73.5-94.1% with coefficients of variation of 4.35-16.60%. The limit of detection (LOD) was 0.002 mg kg(-1) for sulfacetamide, sulfapyridine, sulfamerazine, and sulfamethoxypyridazine; that for sulfameter, sulfachloropyridazine, sulfamethoxazole and sulfadimethoxine was 0.005 mg kg(-1). The limit of quantitation (LOQ) was 0.005 mg kg(-1) for sulfacetamide, sulfapyridine, sulfamerazine, and sulfamethoxypyridazine; that for sulfameter, sulfachloropyridazine, sulfamethoxazole, and sulfadimethoxine was 0.010 mg kg(-1). The method is suitable for determination of multiresidue sulfonamides in the various kinds of honey. PMID- 12739096 TI - Clean-up of triazines in vegetable extracts by molecularly-imprinted solid-phase extraction using a propazine-imprinted polymer. AB - An analytical methodology based on a molecularly imprinted solid-phase extraction (MISPE) procedure was developed for the determination of several triazines (atrazine, simazine, desethylatrazine (DEA), desisopropylatrazine (DIA), and propazine) in vegetable samples. A methacrylic acid-based imprinted polymer was prepared by precipitation polymerisation using propazine as template and toluene as porogen. After removal of the template by Soxhlet extraction, the optimum loading, washing, and elution conditions for MISPE of the selected triazines were established. The optimised MISPE procedure was applied to the extraction of the selected triazines in pea, potato, and corn sample extracts and a high degree of clean-up was obtained. However, some remaining interferences, non-specifically and strongly bound to the polymeric matrix, appeared in the chromatogram, preventing quantification of DIA in potatoes and DIA, DEA, and propazine in corn samples. Thus, a new clean-up protocol based on the use of a non-imprinted polymer for removal of these interferences prior to the MISPE step was developed. By following the new two-step MISPE procedure, the matrix compounds were almost completely removed, allowing the determination of all the triazines selected at concentration levels below the established maximum residue limits, making the developed procedure suitable for monitoring these analytes in vegetable samples. PMID- 12739097 TI - Trace elemental analysis of automotive paints by laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS). AB - Paints and coatings are frequently encountered as types of materials that are submitted to forensic science laboratories as a result of trace evidence transfers. The aim of this study was to develop a method to complement the commonly used techniques in a forensic laboratory in order to better characterize these samples for forensic purposes. A laser ablation method has been used to simultaneously sample several layers directly prior to introduction into an inductively coupled plasma-mass spectrometer for the detection and quantification of the trace metals present in the layer(s). Time-resolved analysis plots displaying the elemental response and quantification of selected metals are compared to associate/discriminate paint samples. Matrix-matched standards were successfully incorporated into the analysis scheme for quantification of lead in the solid paint samples. Preparation of new matrix-matched standards for quantification of additional elements developed for this study are also presented. A sample set of eighteen (18) survey automotive paint samples have been analyzed with the developed method in order to determine the utility of LA ICP-MS for trace element analysis of paints. PMID- 12739098 TI - Quantification of DNA in forensic samples. AB - Quantification of DNA in a forensic sample is of major importance for proper DNA amplification and STR profiling. Several methods have been developed to quantify DNA, from basic UV spectrometry, through gel-based techniques, to dye staining, blotting techniques, and, very recently, DNA amplification methods (polymerase chain reaction, PCR). Early techniques simply measured total DNA, but newer techniques can specifically measure human DNA while excluding non-human DNA (foodstuff, animal, or bacterial contamination). These newer assays can be faster and less expensive than traditional methods, making them ideal for the busy forensic laboratory. This paper reviews classic and newer quantification techniques and presents methods recently developed by the authors on the basis of PCR of Alu sequences. PMID- 12739099 TI - Improved selectivity and detection limit of the carbonate-selective electrode. AB - The properties of the carbonate neutral carrier 4-( n-hexadecyl)-3-nitro-1 trifluoroacetylbenzene were compared with those of a similar carrier, without a nitro group, studied previously. In spite of differences in the Hammett constant of the carbonyl group responsible for interaction with the analyte, the analytical characteristics of both carriers, measured under the same conditions, were comparable. Special care was taken to avoid the presence of an excessive carbon dioxide level in the diffusion layer at the membrane-solution interface. The internal reference solution was prepared without carbonate components; the external solution was protected from contact with atmospheric carbon dioxide. Under such conditions the detection limit of both electrodes was extended to 10( 11 )mol L(-1), and the selectivity towards salicylate, chloride, and acetate was significantly improved. PMID- 12739100 TI - Analytical procedure for the simultaneous voltammetric determination of toxic metals in dialysis fluids. AB - A new analytical voltammetric procedure for the simultaneous determination of copper(II), lead(II), cadmium(II), zinc(II), chromium(VI), and manganese(II) in two kinds of dialysis fluid (peritoneal and haemodialysis fluids) is described. The voltammetric measurements were performed using, as working electrode, a stationary mercury electrode, and a platinum electrode and a Agmid R:AgClmid R:KCl ((sat.)) electrode as auxiliary and reference electrodes, respectively, employing 0.1 mol L(-1) dibasic ammonium citrate solution pH 6.9 as supporting electrolyte. For all the elements, the accuracy, expressed as relative recovery R%, was very satisfactory being in the range 94-105%, the precision, expressed as relative standard deviation s(r)%, was lower than 6%, while the limits of detection were of the order of a few units of microg L(-1). The analytical voltammetric procedure has been validated by comparison with spectroscopic (graphite furnace atomic absorption spectroscopy, GFAAS) measurements. PMID- 12739101 TI - Morin applied in speciation of aluminium in natural waters and biological samples by reversed-phase high-performance liquid chromatography with fluorescence detection. AB - A reversed-phase high-performance liquid chromatographic method with fluorescence detection for the determination of labile monomeric aluminium has been developed through pre-column complexation using morin as the analytical reagent. The highly fluorescent aluminium-morin complex (excitation wavelength 418 nm, emission wavelength 490 nm) was separated on a Spherisorb ODS 2 column with an eluent consisting of 30% methanol and 70% water (pH 1.0 with perchloric acid). The most remarkable point of this protocol was that only the most toxic aluminium species, that is, free aqua-aluminium ion and its monomeric hydroxo complex ions, selectively respond among various aluminium complexes. This strategy has been successfully applied to direct fractionation of the toxic aluminium in natural waters and biological samples without any pretreatment. PMID- 12739102 TI - XRCC1, CYP2E1 and ALDH2 genetic polymorphisms and sister chromatid exchange frequency alterations amongst vinyl chloride monomer-exposed polyvinyl chloride workers. AB - Vinyl chloride monomer (VCM) is a known human carcinogen, which may be metabolized by cytochrome P450 2E1 (CYP2E1), aldehyde dehydrogenase 2 (ALDH2), and glutathione S-transferase T1 (GSTT1). A DNA-repair gene, X-ray repair cross complementing group 1 ( XRCC1, exon 10), may also be implicated in the process of VCM-related carcinogenesis. Thus, VCM-exposed workers with inherited susceptible metabolic and DNA-repair genotypes may experience an increased risk of genotoxiciy. This study was designed to investigate whether metabolic and DNA repair genotypes affected sister chromatid exchange (SCE) frequency in occupationally VCM-exposed workers from polyvinyl chloride (PVC) manufacturing plants. Study subjects comprised 61 male workers having experienced VCM exposure, and 29 male controls. Questionnaires were administered to obtain detailed histories of cigarette-smoking habits, alcohol consumption behavior, and occupation. The frequency of SCE in peripheral lymphocytes was determined using a standardized method, and genotypes of CYP2E1, ALDH2, GSTT1 and XRCC1 were identified by the polymerase chain reaction (PCR) procedure. Our results demonstrated that smoking, age and VCM exposure and XRCC1 ( P=0.03), CYP2E1 ( P=0.04), and ALDH2 ( P=0.08) were significantly associated with an increased SCE frequency. Further analysis of gene combinations, including CYP2E1, ALDH2 and XRCC1, revealed an increased trend for these genotypes to influence SCE frequencies for the low VCM-exposure group ( P<0.01), but not so for the high VCM exposure group ( P=0.29) or for controls ( P=0.49). These results suggest that workers with susceptible metabolic and DNA-repair genotypes, may experience an increased risk of DNA damage elicited by VCM exposure. PMID- 12739103 TI - Mercury pretreatment selects an enhanced cadmium-accumulating phenotype in Euglena gracilis. AB - Pre-treatment of heterotrophic cultures of Euglena gracilis with 1.5 microM HgCl(2) for at least 60 generations resulted in a cell population that showed both increased resistance to Cd(2+) and ability to accumulate it, when compared to non-Hg(2+)-pretreated Euglena. These Hg(2+)-enhanced capacities were evident in cells cultured in the dark in a medium with lactate, but not in cells cultured with glutamate plus malate. After culturing with 0.1 mM CdCl(2) through three consecutive transfers, the mercury-pretreated cells still grew and maintained high levels of glutathione-related metabolites, while the non-Hg(2+)-pretreated cells died. Cultures of Hg(2+)-pretreated cells, after transfer to media with or without cadmium, did not alter either their enhanced Cd(2+) accumulation or their increased production of glutathione-related metabolites. These observations suggested that the Hg(2+)-pretreated population underwent a permanent change that improved its Cd(2+) resistance. Several factors that contributed to the improved capacities included: (a) higher cellular malate, cysteine and glutathione levels induced by Hg(2+) before and after Cd(2+) exposure; and (b) increased storage of Cd(2+) in mitochondria along with increased intramitochondrial citrate, cysteine, and glutathione levels. These characteristics suggested that this Cd(2+) hyper accumulating strain of E. gracilis might be a suitable candidate for Cd(2+) bioremediation of polluted water systems. PMID- 12739105 TI - Culture collections and biochemistry. AB - This review describes the relationships and links between culture collections, which act as sources of genomes, transcriptomes, proteome, and metabolomes, and fields of research biochemistry that demand their support and help. In addition, the invaluable but not always rewarded efforts of these organizations as a source and conservator of organism diversity is discussed. Biological waste-water treatment, ethanol as a non-finite source of energy, Rhodococcus fascians as the source of a citrus-juice debittering agent, the sporulation of filamentous fungi in liquid medium, and biotransformation with growing and resting cells are processes developed by the authors that demonstrate some of the applications of organisms from culture collections in the general field of biotechnology and related areas, including industrial biochemistry and biocatalytic synthesis. PMID- 12739106 TI - Failure to detect Chlamydia pneumoniae by cell culture and polymerase chain reaction in major arteries of 93 patients with atherosclerosis. AB - To detect Chlamydia pneumoniae in punch specimens of the aortic wall of 61 patients undergoing coronary-aortic bypass graft, and carotid atheromas of 32 patients undergoing carotid endarterectomy, cell culture (HEp-2 cells) and two polymerase chain reaction assays in two different laboratories were used. All cultures and polymerase chain reaction tests for Chlamydia pneumoniae were negative. Further studies are required to explore the complex relationship between Chlamydia pneumoniae and atherosclerosis. PMID- 12739104 TI - Genetic variation of circulating leptin is involved in genetic variation of hand bone size and geometry. AB - Leptin is secreted primarily by the adipocytes and plays an important role in the regulation of food intake and energy expenditure. In addition to its adipostatic function, it has been demonstrated that leptin directly enhances stromal cell differentiation to osteoblasts, and since such precursor cells are potential targets for leptin, the latter could possibly mediate the relationship between obesity and bone mass and size. To address this question, we studied phenotypic and genetic correlations between the circulating levels of leptin and hand bone size (BS) and geometry (BG) of the radiographic hand in a healthy and ethnically homogeneous sample of pedigrees. We also attempted to evaluate to what extent potential leptin/BS/BG correlations are modified by an individual's obesity traits, specifically his/her BMI. Our research has shown that leptin, BMI and the corresponding bone measures are clearly inherited traits (0.46+/-0.11, 0.35+/ 0.16, 0.62+/-0.12 and 0.51+/-0.09, respectively). The bivariate variance component analysis revealed very strong and significant genetic and environmental correlations between circulating leptin and BMI ( r(G)=0.86+/-0.09, r(E)=0.75+/ 0.05, P<0.001). Furthermore, genetic correlations between leptin and hand bone characteristics proved inverse and statistically significant ( r(G)=-0.35+/-0.01 and -0.45+/-0.10 for BS and BG, respectively), while corresponding environmental correlations were low ( r(E)=-0.14+/-0.15 and -0.07+/-0.14) and they could be constrained to zero without significant deterioration of the model fit to the data ( P>0.10). However, despite the extremely strong relationship between leptin and BMI, we failed to detect phenotypic or genetic correlations between BMI and our two hand bone measures. Thus our study provided evidence that plasma leptin levels may be statistically significant predictor of hand bone size and geometry, and may play a physiological role in maintaining bone mass as well as in regulation of hand bone proportions. PMID- 12739107 TI - Non-fatal acute liver injury possibly related to high-dose ciprofloxacin. AB - Fulminant hepatic failure is an uncommon but life-threatening adverse reaction to antibiotic therapy. Reported here is a case of severe liver injury associated with ciprofloxacin administered for treatment of a gram-negative infection in a 79-year-old female. The patient presented with metabolic acidosis 48 hours after the first ciprofloxacin intake. Symptoms resolved after cessation of ciprofloxacin therapy. This report highlights an uncommon side effect of a commonly used antimicrobial agent. PMID- 12739108 TI - DNA cleavage studies of mononuclear and dinuclear copper(II) complexes with benzothiazolesulfonamide ligands. AB - Copper-based transition metal complexes performing single- and double-strand scission of DNA have been studied. The dinuclear complexes [Cu(2)(L)(2)(OCH(3))(2)(NH(3))(2)] and [Cu(2)(L)(2)(OCH(3))(2)(DMSO)(2)] are more active than the corresponding mononuclear [Cu(L)(2)(py)(2)] (where HL= N-(4 methylbenzothiazol-2-yl)benzenesulfonamide), suggesting that the dinuclearity is an important factor in the oxidative cleavage of DNA. The cleavage efficiency of the complexes depends on the reducing agent used in the process, the tandem ascorbate/H(2)O(2) being the most efficient. PAGE analyses have shown that these complexes cleave DNA without sequence selectivity. The DNA degradation process takes place mainly by C1' oxidation, but C4' and C5' oxidations cannot be ruled out as minor pathways. These copper complexes preferably oxidize guanine under mild conditions, but under more drastic conditions the oxidation reactivity appears to be T>G>C>A, suggesting the intervention of hydroxyl radicals as active species. PMID- 12739109 TI - Tourism climatology: evaluating environmental information for decision making and business planning in the recreation and tourism sector. AB - This work grew from initiatives of the International Society of Biometeorology's Commission on Climate, Tourism and Recreation (ISBCCTR). The ISBCCTR was formed during the 15th Congress of the ISB held in November 1999 in Sydney, Australia. The aim of ISBCCTR is to promote research in tourism climatology. The first formal meeting of the Commission took place at the Meliton Resort, Halkidiki, Greece, 5-10 October 2001. The aims were to (1) bring together a selection of scientists and tourism experts to review the current state of knowledge of tourism climatology and (2) explore areas and priorities for future work and the role of the Commission in this. The Workshop highlighted the fact that, although climate is widely recognised as vitally important to tourism, relatively little is known about its effects. Even less is known about the economic impact or significance of climate on commercial prospects for tourism. Important research themes that warrant attention were identified. Among these was the need for a tourism climate index (or indices) that integrates all facets of climate, uses standard data and is objectively tested and verified. Work is also required on developing a better understanding of what climate-related information is required by both tourists and the tourism industry, exploring the distinction between the impact of climate on tourists and the impact on the tourism industry, setting a standard approach to tourism climate assessment, assessing the role of weather forecasts and long-term expectations of climate on choices made by tourists, the risks to tourism caused by extreme atmospheric events, what climate-related criteria people use to make decisions about tourism and recreational choices, how products giving information about weather and climate are currently used by the recreation and tourism industry and what are the existing and future requirements for this climate information. PMID- 12739111 TI - Percutaneous fetoscopic patch coverage of experimental lumbosacral full-thickness skin lesions in sheep. AB - BACKGROUND: In order to minimize maternal trauma from open fetal surgery for prenatal coverage of fetal myelomeningoceles in humans, we assessed the feasibility of a percutaneous fetoscopic approach in sheep. METHODS: In seven ewes between 90 and 100 days of gestation, the amniotic cavity was entered percutaneously. Each fetus was postured and a full-thickness skin lesion was created in the lumbosacral region. Then, the feasibility of covering this lesion with a patch and fetal skin by standard endoscopic suturing techniques (n = 3) or robot assistance (n = 4) was assessed. RESULTS: Percutaneous fetoscopic patch and skin coverage of the full-thickness skin lesion was achieved in six of the seven fetal sheep. Five fetuses survived gestation and were delivered healthy. CONCLUSIONS: Percutaneous fetoscopic posturing and patch coverage of lumbosacral full-thickness skin lesions can effectively and safely be achieved in sheep. This approach promises to provide a substantial reduction of maternal trauma from fetal surgery for myelomeningoceles. PMID- 12739112 TI - Distribution of intrasplenically injected colon cancer cells following pneumoperitoneum in mice. AB - BACKGROUND: Few studies have examined tumor cell distribution following laparoscopic surgery for colorectal cancer. We examined the effect of carbon dioxide pneumoperitoneum on the distribution of intrasplenically injected colon cancer cells in mice. METHODS: Mice were intrasplenically injected with 2 x 10(4) colon 26 cells labeled with 111In-oxine and were randomized to undergo pneumoperitoneum at 10 mmHg for 30 min or to receive no treatment other than anesthesia. Radioactivity of the liver, lungs, and spleen was measured 30, 60, 90, or 150 min following tumor inoculation. RESULTS: The dynamic changes in the hepatic radioactivity were not similar between groups. However, the values were not significantly different at any time point. The radioactivity of lungs was extremely low in both groups throughout the experimental period. CONCLUSIONS: Pneumoperitoneum does not appear to cause the accumulation of intraportally spreading tumor cells in the liver, but it may affect the dynamic changes of tumor cells. Also, tumor cell localization in the lungs is negligible in both pneumoperitoneum and control groups. PMID- 12739113 TI - A nonsurgical technique to create an esophagostomy for difficult cases of percutaneous endoscopic gastrostomy. AB - BACKGROUND: We developed a minimally invasive technique of esophagostomy called percutaneous transesophageal gastrotubing (PTEG) using a rupture-free balloon (RFB) for enteral nutrition and drainage as well as percutaneous endoscopic gastrostomy (PEG). PTEG using RFB allows surgeons to create a nonsurgical esophagostomy even in difficult cases of PEG (i.e., total gastrectomized patients and massive ascites). METHODS: To create the PTEG, a RFB is inserted into the esophagus through the nose and inflated. The RFB is punctured with a needle at the left neck under ultrasonographic vision. A guidewire is inserted through the needle, followed by dilatation of the punctured site using a dilator with sheath. Finally, the tube is inserted into the gastrointestinal tract and the sheath is peeled off. RESULTS: From January 1998 to June 2002, we treated 115 patients using PTEG with a RFB and there were no major complications. Therapeutic results are as good as those for PEG and it took approximately 15 min to perform. CONCLUSIONS: PTEG with RFB is as safe as PEG and is simple and less invasive. It can be used in some cases for which PEG is contraindicated. PMID- 12739115 TI - Transanal endoscopic excision of rectal adenomas. AB - Transanal endoscopic microsurgery (TEM) is a minimally invasive surgical technique for performing local excision of rectal lesions in the mid and upper rectum that would otherwise be inaccessible for local excision by the direct transanal approach. In the absence of this approach, low anterior resection would be required, which is major abdominal surgery. The justification for excising adenomas of the colon and rectum is their malignant potential, which correlates with the size of the lesion. This retrospective review examines our experience using TEM for excision of adenomas of the rectum from February 1991 to the present. The decision for using TEM is based on a precise localization of the lesion with particular attention to the upper margin of the lesion and its diameter. A total of 56 adenomas were removed. The average diameter was 4.9 cm (range, 3-8 cm). The average distance from the anal verge was 7.92 cm (range, 5 12 cm). Carcinoma in situ was seen in 7 lesions, and the remaining lesions were benign. Morbidity was minimal, with one conversion to an open procedure for an intraperitoneal perforation that required a low anterior resection. No patient required transfusion and there was no mortality. The hospital stay was short, with half of the patients being discharged the same day. The average cost from July 1996 to December 1999 was 7775 dollars for TEM versus 34,018 dollars for LAR. Subsequent follow-up average was 38.8 months (range, 1-100 months), during which time two patients had recurrence of their adenomas. This was successfully treated with reexcision. In conclusion, TEM is an accurate, safe, and relatively inexpensive technique when compared to low anterior resection. This technique significantly reduces the proportion of adenomas requiring abdominal surgery. PMID- 12739114 TI - Long-term results from laparoscopic common bile duct exploration. AB - BACKGROUND: The aim of this study was to evaluate the long-term results after laparoscopic common bile duct exploration (LCBDE). METHODS: A retrospective review of 175 consecutive patients who underwent attempted LCBDE between 1992 and 1999 was conducted. Laparoscopic transcystic exploration was accomplished in 110 patients and laparoscopic choledochotomy in 52 patients. Conversion to an open common bile duct exploration was required for 13 patients (7.4%). Retained common bile duct stones occurred in eight patients (4.6%). The 30-day postoperative morbidity was 6.9%, and there was no 30-day mortality. All the patients (alive and localized) received a questionnaire evaluating long-term results. RESULTS: Of the 175 patients, 169 (4 unrelated deaths and 2 patients lost to follow-up evaluation) received and 152 (90%) returned the questionnaire. The follow-up period ranged from 6 to 72 months (median, 36 months). One patient developed recurrent common bile duct stones. There were no signs or evidence of common bile duct stricture in any patient. CONCLUSION: The LCBDE procedure can be performed without increased risk of late bile duct complications. PMID- 12739117 TI - Effectiveness of laparoscopic fundoplication in relieving the symptoms of gastroesophageal reflux disease (GERD) and eliminating antireflux medical therapy. AB - BACKGROUND: Recent reports have suggested that antireflux surgery should not be advised with the expectation of elimination of medical treatment. We reviewed our results with laparoscopic fundoplication as a means of eliminating the symptoms of gastroesophageal reflux disease (GERD), improving quality of life, and freeing patients from chronic medical treatment for GERD. METHODS: A total of 297 patients who underwent laparoscopic fundoplication (Nissen, n = 252; Toupet, n = 45) were followed for an average of 31.4 months. Preoperative evaluation included endoscopy, barium esophagram, esophageal manometry, and 24-h pH analysis. A preoperative and postoperative visual analogue scoring scale (0-10 severity) was used to evaluate symptoms of heartburn, regurgitation, and dysphagia. A GERD score (2-32) as described by Jamieson was also utilized. The need for GERD medications before and after surgery was assessed. RESULTS: At 2-year follow-up, the average symptom scores decreased significantly in comparison to the preoperative values: heartburn from 8.4 to 1.7, regurgitation from 7.2 to 0.7, and dysphagia from 3.7 to 1.0. The Jamieson GERD score also decreased from 25.7 preoperatively to 4.1 postoperatively. Only 10% of patients were on proton pump inhibitors (PPI) at 2 years after surgery for typical GERD symptoms. A similar percentage of patients (8.7%) were on PPI treatment for questionable reasons, such as Barrett's esophagus, "sensitive" stomach, and irritable bowel syndrome. Seventeen patients (5.7%) required repeat fundoplication for heartburn ( n = 9), dysphagia ( n = 5), and gas/bloating ( n = 3). CONCLUSIONS: Laparoscopic fundoplication can successfully eliminate GERD symptoms and improve quality of life. Significant reduction in the need for chronic GERD medical treatment 2 years after antireflux surgery can be anticipated. PMID- 12739116 TI - Predicting conversion in laparoscopic colorectal surgery. Fellowship training may be an advantage. AB - BACKGROUND: Laparoscopic colorectal surgery has clear advantages over open surgery; however, the effectiveness of the approach depends on the conversion rate. The objective of this work was to prospectively validate a model that would predict conversion in laparoscopic colorectal surgery. METHODS: A simple clinical model for predicting conversion in laparoscopic colorectal surgery was previously developed based on a multivariable logistic regression analysis of 367 procedures. This model was applied prospectively to a follow-up group of 248 procedures by the same team, including 54 procedures performed by one new fellowship-trained surgeon. RESULTS: Patients in the follow-up group were more likely to have cancer (56% vs 44%, p = 0.007) and were more obese (median, 71.0 vs 66.0 kg; p < 0.001). The rate of conversion in the follow-up group was unchanged (8.9% vs 9.0%, p > 0.05). Despite expected trends toward increasing risk of conversion with weight level (<60 kg, 6.8%; 60-<90 kg, 9.0%; >90 kg, 12.1%; p > 0.05) and malignancy (10.1% vs 7.3%, p > 0.05), the model did not distinguish well between groups at risk for conversion. Contrary to the model, however, the fellowship-trained surgeon had a conversion rate that was not higher than that of the other, more experienced surgeons (7.3% vs 9.3%, p > 0.05) even though he was less experienced, and operating on patients who were more obese (median, 75.0 vs 70 kg; p = 0.02) and more likely to have cancer (59% vs 55%, p > 0.05). Recalculated conversion scores that excluded the inexperience point for the fellowship-trained surgeon showed a good fit for the model. Considering the original and follow-up experience together (615 cases), the model clearly stratifies patients into low (0 points), medium (1-2 points), and high risk (3-4 points) for conversion, with respective rates of 2.9%, 8.1%, and 20% ( p = 0.001). CONCLUSION: This model appears to be a valid predictor of conversion to open surgery. Fellowship training may provide sufficient experience so that learning curve issues are redundant in early practice. This model now requires validation by other centers. PMID- 12739119 TI - Transanal endoscopic microsurgery and radical surgery for T1 and T2 rectal cancer. AB - BACKGROUND: Transanal endoscopic microsurgery (TEM) has gained increasing acceptance as a local treatment of early rectal cancer. The purpose of this study was to compare the results of TEM and radical surgery in patients with T1 and T2 rectal cancer. METHODS: From October 1994 to December 2000, 74 patients with T1 and T2 rectal adenocarcinoma treated with TEM were compared with 100 patients with T1N0M0 and T2N0M0 rectal adenocarcinoma treated with radical surgery. Retrospective analysis was performed regarding to recurrence and survival rate. Neither group received adjuvant chemoradiation. There was no significant difference in age, gender, tumor location, or follow-up period between the two groups. The only difference was in tumor size. RESULTS: Of the 74 patients in TEM group, 52 were T1 (70.3%) and 22 were T2 (29.7%). Of the 100 patients in radical surgery group, 17 were T1 (17%) and 83 patients were T2 (83%). The 5-year local recurrence rates were 4.1% for T1, 19.5% for T2 after TEM, 0% for T1, and 9.4% for T2 after radical surgery. There was no statistical difference between the TEM and radical surgery groups for T1 rectal cancer ( p = 0.95), but for T2 rectal cancer, the 5-year local recurrence rate was higher after TEM than after radical surgery ( p = 0.04). There were no significant statistical difference between the two groups in terms of the 5-year disease-free survival rate and the survival rate. CONCLUSIONS: For T1 rectal cancer, there was no difference in recurrence or 5-year survival rate between the TEM and the radical surgery groups. For T2 rectal cancer, there was no statistical difference in the 5-year survival rate between the two groups, but TEM carried higher risk of local recurrence. Therefore, careful selection of the patients is required for TEM, and when proper muscle invasion is proven, the TEM procedure should be supplemented by further treatment, or radical surgery should be performed. PMID- 12739120 TI - Pitfalls and complications with laparoscopic intraperitoneal expanded polytetrafluoroethylene patch of postoperative ventral hernia. PMID- 12739118 TI - Incidence and management of Mirizzi syndrome during laparoscopic cholecystectomy. AB - BACKGROUND: Benign extrinsic obstruction of the hepatic duct, known as "Mirizzi syndrome" (MS), is an uncommon complication of longstanding cholelithiasis. Since laparoscopic cholecystectomy (LC) replaced the open approach, Mirizzi syndrome has regained the interest of biliary surgeons. METHODS: The Swiss Association for Laparoscopic and Thoracoscopic Surgery (SALTS) prospectively collected the data on 13,023 patients undergoing LC between 1995 and 1999. This database was investigated with special regard to patients with Mirizzi syndrome. RESULTS: There were 39 patients (14 men and 25 women; mean age, 61 years) with MS (incidence, 0.3%). Thirty-four patients had type 1 MS and five had type 2. A gallbladder carcinoma was found in four patients (incidence, 11%). In the type 1 group, 23 patients underwent cholecystectomy only, 10 patients had a bile duct exploration and T-tube insertion, and one patient had a Roux-en-Y reconstruction. In three patients with type 2, a hepaticojejunostomy was performed; two others underwent simple closure and drainage (via T-tube) of the biliary fistula. The conversion rate was 74% (24 of 34 patients) in the type 1 group and 100% (five of five patients) for type 2. The overall complication rate was 18%. There were no deaths. CONCLUSIONS: Although MS is rarely encountered during LC, it must be recognized intraoperatively. Conversion to an open approach is often needed, and prior to any surgical intervention, gallbladder cancer must be excluded. PMID- 12739121 TI - Optimization of the projection screen in a display system for minimal access surgery. AB - BACKGROUND: The operative image for minimal access surgery currently is displayed on a monitor located outside the sterile field. It is ergonomically advantageous to locate the image adjacent to the surgeon's hands by projection onto a sterile screen, but there has been no research into the optimal screen material. METHODS: Several screen materials were compared for image resolution, brightness, variation of brightness with viewing angle, and image artifact. RESULTS: Glossy materials perform poorly, whereas finely grained surfaces improve image clarity. Excessive roughness and incomplete opacity limit the image resolution. Conventional screen fabrics are unsuitable in this application. Ambient lighting and projector brightness affect image contrast, but a correct choice of material can address this. Practical issues such as moisture absorbency and ease of sterilization are considered. CONCLUSIONS: Potential screen materials were rejected because of excessive glare, poor resolution, and image artifact. Finely textured surfaces (e.g., polystyrene sheeting) provide an acceptable screen. PMID- 12739122 TI - Combining ultrasonic dissection and the Storz operation rectoscope. AB - BACKGROUND: Transanal endoscopic microsurgery (TEM) allows a precise, full thickness resection of rectal tumors anywhere within the rectum. Unfortunately, the standard TEM technique needs complex and rather expensive equipment, demands high skill, and is attended by bleeding and oozing that may be challenging. A modified TEM procedure combining the new Storz operation rectoscope and ultrasonic dissection has been developed to overcome the limitations of the original technique. METHODS: The Storz operation rectoscope features a 5-mm telescope combined with a single-monitor display. Standard laparoscopic instruments and the LCSC5 Ultracision Maniple are used for dissection and coagulation. Full-thickness resection is performed most often. Closure of the defect is accomplished by interrupted 3-0 polydoxanone sutures secured by extracorporeal slipknots. RESULTS: Altogether, 18 TEMs have been performed according to the modified technique: 9 for malignant and 9 for benign lesions. The median operating time was 92.5 min for resection of malignant lesions and 40 min for resection of benign lesions. Two postoperative complications occurred: a bleeding and a partial dehiscence. The median follow-up periods were 35 months for malignant disease and 19.5 months for benign disease. No recurrence was observed. CONCLUSION: For tumors located up to 15 cm from the anal verge, TEM with the Storz rectoscope and ultrasonic dissection is indicated. Despite the complication described, coagulation is optimal and ultrasonic scissors allow working in a fairly bloodless field. The overall costs of the equipment are significantly lower. PMID- 12739123 TI - The utility of laparoscopy in the diagnosis of acute appendicitis in women of reproductive age. AB - AIM: To see whether laparoscopy improves the accuracy of a clinical diagnosis of acute appendicitis in women of reproductive age, and to determine what the long term sequelae are of not removing an appendix deemed at laparoscopy to be normal. METHOD: The initial part of the study was undertaken during 1991-1992. Female patients between 16 and 45 years were eligible for inclusion once a clinical decision had been made to perform an appendicectomy for suspected acute appendicitis. Following consent, patients were randomized into two groups. One group had open appendicectomy, as planned. The other group had laparoscopy, followed by open appendicectomy only if the appendix was seen to be inflamed or was not visualized. The end points for the study were the clinical outcomes of all patients, and the results of histology, where appropriate. An attempt was made to contact all patients at 10 years to determine whether they had had a subsequent appendicectomy, or had been diagnosed with another abdominal condition that might be relevant to the initial presentation in 1991-1992. RESULTS: Laparoscopic assessment was correct in all cases in which the appendix was visualized. Diagnostic accuracy was improved from 75% to 97%. Laparoscopy was associated with no added complications, no increase in hospital stay in patients who went on to appendicectomy, and a reduction in hospital stay for those who underwent laparoscopy alone. No patients developed a problem over the 10-year follow-up period from having a normal-looking appendix not removed at laparoscopy. CONCLUSION: Laparoscopic assessment of the appendix is reliable, and to leave a normal-looking appendix at laparoscopy does not appear to cause any long-term problems. PMID- 12739124 TI - Histologic characteristics of laparoscopic saline-enhanced electrosurgery of liver and splenic injuries. AB - BACKGROUND: Hemostasis is crucial during laparoscopic surgery. Unlike conventional electrosurgery, saline-enhanced electrosurgery (SEE) improves the electrode-to-tissue interface. This study investigated SEE as a means of achieving hemostasis in liver and splenic injuries and assessed the associated histologic changes. METHODS: Nine anesthetized pigs underwent laparoscopic liver tip and splenic wedge amputations. Injuries were treated with SEE (25-35 and 35 45 W, respectively). Animals were sacrificed postprocedure and at 1 and 4 weeks. Depth of necrosis was analyzed using a mixed model analysis of variance. RESULTS: Liver mean depth of necrosis was 5.3 mm (acute), 6.0 mm (1 week), and 5.3 mm (4 weeks) ( p = not significant). Splenic mean depth of necrosis was 7.0 mm (acute), 7.0 mm (1 week), and 2.7 mm (4 weeks). ( p < 0.01) Acute injuries demonstrated thermal coagulation necrosis that healed with well-defined bands of collagen at 1 and 4 weeks. CONCLUSIONS: SEE provides effective hemostasis and results in an acceptable depth of necrosis with satisfactory wound healing in a porcine model of solid organ injury. PMID- 12739126 TI - Gasless, video endoscopic transanal excision for carcinoid and laterally spreading tumors of the rectum. AB - BACKGROUND: The aim of this study is to determine whether gasless, video endoscopic transanal-rectal tumor excision (gasless VTEM) is a valid treatment for rectal carcinoid and laterally spreading tumors (LST). METHODS: Eighty-four patients with an adenoma, adenocarcinoma (Tis/T1), or carcinoid tumor of the rectum were divided into three groups: (i) LST ( n = 17 patients), (ii) carcinoid ( n = 11), and (iii) control with other types of tumors ( n = 56). RESULTS: The LST group had a longer median operating time than in the control group, whereas the carcinoid group had a shorter operating time. Two patients (11.7%) in LST group developed peritoneal entry during the operation, while 2 patients (3.6%) in the control group experienced postoperative complications. During a median follow up length of 55.2 months, one patient in the LST group developed a recurrence. CONCLUSIONS: Gasless VTEM is a simple, minimally invasive procedure used to treat LST and carcinoid tumors of the rectum. However, resection for the LST group had a high risk of peritoneal entry during operation. PMID- 12739125 TI - Nissen fundoplication has a vagolytic effect on the lower esophageal sphincter. AB - BACKGROUND: Laparoscopic Nissen fundoplication is an increasingly utilized option for the treatment of gastroesophageal reflux disease (GERD). However, many questions remain as to the mechanism by which this operation prevents GERD in those without hiatal hernias or incompetent lower esophageal sphincters (LES). It is known that these patients experience reflux due to excess transient lower esophageal sphincter relaxations (TLESR), inappropriate and short-lived relaxation of the LES and crural diaphragm. The purpose of this study was to determine if Nissen fundoplication affects the neural pathways involved in the TLESR reflex. METHODS: Five dogs were anesthetized and intubated. Both vagal nerves and the right phrenic nerve were isolated. A continuous water perfusion manometric catheter was situated at the LES. The nerves were then electrically stimulated and the resultant pressure at the LES measured at baseline, and during and after an open Nissen fundoplication. RESULTS: The mean LES pressures before dissection, after esophago-gastric mobilization, and after fundoplication were 47 +/- 13 mmHg, 21 +/- 9 mmHg, and 14 +/- 4 mmHg, respectively. All differences were significant. There was no change noted in LES pressure with stimulation of either or both of the phrenic nerves without concomitant vagal stimulation. CONCLUSION: Nissen fundoplication may prevent GERD in those without a hiatal hernia or incompetent LES by disrupting the efferent vagal fibers to the LES. Such fibers mediate TLESR which are responsible for GERD in these patients. PMID- 12739127 TI - Laparoscopic treatment of Bochdalek hernia without the use of a mesh. AB - Bochdalek hernia is a rare pathology. The preoperative diagnosis is difficult, and few reports are available regarding its treatment. Herein we report the case of a 25-year-old woman referred for symptoms of dyspepsia, dysphagia, and thoracic pain exacerbated by pregnancy. Preoperative radiography, EGD, and CT scan revealed a paraesophageal hiatal hernia. Laparoscopic exploration showed the complete thoracic migration of the stomach through a left posterolateral diaphragmatic foramen. The diagnosis of a Bochdalek hernia was then made. The diaphragmatic defect was repaired without inserting a prosthesis, using five separate non-reabsorbable stitches (Rieder technique). The procedure was completed with a Nissen-Rossetti fundoplication. The duration of the procedure was 150 min. Hospital stay was 12 days. There were no complications. Postoperative Gastrografin radiography of the esophagus and stomach showed a normal-shaped fundoplication and confirmed the subdiaphragmatic location of the stomach. We conclude that the laparoscopic approach represents the gold standard for the diagnosis and treatment of Bochdalek hernia and any associated complications. PMID- 12739128 TI - Impact of laparoscopy on the management of an unusual case of nonpalpable testis in an adult patient. AB - BACKGROUND: In the past decade, cryptorchidism has generally been treated in the first 36 months of life using an inguinal approach, in the case of palpable testis, or using laparoscopy, in the case of nonpalpable testis. Nevertheless, before this period some children were managed incorrectly. This case report shows how laparoscopy and the collaboration between pediatric surgeons and urologists may lead to optimal results. CASE REPORT: A 19-year-old male was referred to our unit following routine medical examination for enrollment in the military service with a diagnosis of right cryptorchidism. The clinical history showed the anamnesis of right cryptorchidism diagnosed at birth as a right nonpalpable testis, confirmed at 2 years of age with clinical examination and computer tomography. The patient came to our attention to again undergo a clinical examination, ultrasonography, computed tomography, and magnetic resonance imaging. We found a left, well-positioned testis of 20 ml in volume and an empty right scrotum; all the instrumental examinations were negative. The patient was thus scheduled for a diagnostic laparoscopy. A pediatric surgeon with extensive experience in this pathology performed the procedure. The 10-mm 30 degrees optics introduced through the umbilicus showed a closed right inguinal ring, with no evidence of either vas deferens or inner spermatic vessels at the level of the inguinal region or pelvis. Two more trocars were introduced and a testis of 15 ml in volume was found under the ascending colon 10 cm away from the cecum. An orchiectomy was performed via laparoscopy. CONCLUSION: This case clearly shows that some males with nonpalpable testis may have been treated incorrectly in the prelaparoscopic era and may now have an intraabdominal testis. In addition, our experience shows that intraabdominal testis may sometime be in an unusual location, and a laparoscopic surgeon with experience in this pathology is fundamental to finding the testis. In the case of adults with nonpalpable testis, when echography, computed tomography, and magnetic resonance imaging are not useful, the only effective diagnostic procedure is laparoscopy. PMID- 12739129 TI - Use of multislice helical computed tomography cholangiography in the diagnosis of biliary disease. AB - A recent advance in computed tomography (CT) technology, multislice helical CT, has enabled production of clearer three-dimensional (3D) images and has drawn interest. We report the usefulness of CT cholangiography using a multislice helical CT scanner for the diagnosis and preoperative imaging of the biliary duct in a case of peculiarly shaped gallbladder with cholecystitis. A 34-year-old woman admitted to our hospital presented with chronic hypochondralgia. A CT scan showed that the gallbladder was normal without wall thickening or stones. However, there appeared to be a tumor, containing a stone approximately 1 cm in diameter, attached under the gallbladder in front of the right kidney and extending up to its lower level. Magnetic resonance cholangiography also depicted a normal gallbladder without wall thickening or stones. Hence, gallbladder stones were not diagnosed by previously the mentioned investigations. In contrast, a 3D image produced by multislice helical CT cholangiography was very clear. From the bottom of the gallbladder, a narrow canal continued to a stone. We diagnosed that the wall of the lower part of the long gallbladder had become thick and elongated because of chronic cholecystitis caused by a gallbladder stone, and laparoscopic cholecystectomy was performed. Macroscopically, the resected gallbladder showed an extremely thickened wall from the lower body to the fundus, in which a stone was located in the center. Multislice helical CT cholangiography has the potential to become one of the most significant examinations for diagnosis and anatomical analysis of biliary disease prior to laparoscopic cholecystectomy. PMID- 12739130 TI - Phenotype-dependent synthesis of transferrin receptor in rat alveolar epithelial cell monolayers. AB - The iron carrier protein transferrin plays a prominent antioxidant and anti bacterial role in the lower respiratory tract and is present at elevated concentrations in lung epithelial lining fluid relative to plasma. The level of transferrin receptor synthesis in primary cultures of rat alveolar epithelial cells (AECs) was investigated. Transferrin receptor was found to be synthesized early in AEC cultures with the alveolar type II cell-like phenotype. Cell-surface receptor localization was attenuated upon apparent transdifferentiation to the alveolar type I cell-like phenotype later in culture. Binding of (125)I-labeled transferrin to the receptor indicated that surface and total cellular transferrin receptor levels were decreased in the type I-like cells. Inclusion of keratinocyte growth factor (KGF) in culture media (10 ng/ml) resulted in retention of transferrin receptor localized to the basolateral surface. Transferrin-receptor-specific internalization of (59)Fe-transferrin was also limited to the basolateral surface of KGF-treated monolayers. These data suggest that alveolar type II (but not type I) cells express functional transferrin receptor in adult rat alveolar epithelium. PMID- 12739131 TI - First detection of microsporidia in dairy calves in North America. AB - Fecal specimens were obtained from a total of 413 dairy calves from farms in Vermont, New York, Pennsylvania, Maryland, Virginia, North Carolina, and Florida. After removal of fecal debris by sieving and density gradient centrifugation, specimens were examined by fluorescence microscopy, polymerase chain reaction (PCR), and DNA sequencing analysis for the presence of microsporidia. Microscopic examination revealed no spores. PCR using generic primers for microsporidia revealed 70 positive calves. PCR was then conducted using specific primers for Enterocytozoon bieneusi, the most frequently found microsporidian in human infections. These primers revealed 13 positive calves from six farms in five states. DNA sequencing analysis of the 13 E. bieneusi-positive specimens confirmed the PCR results and indicated 96.8-99.8% similarity with E. bieneusi sequences in GenBank. This is the first report of E. bieneusi in cattle in North America. PMID- 12739132 TI - Effect of ambient salinity on immobilization of Caenorhabditis elegans by nematocidal agents. AB - The ability of three antinematodal agents to induce paralysis of Caenorhabditis elegans was examined in an aqueous medium with and without the addition of salts. The basic medium was deionized water supplemented, for control purposes, with the phosphate mixture used as a buffering agent in M9 solution. This medium was further supplemented with magnesium sulfate or sodium chloride, or both salts, at the concentrations used in M9. We report that the paralyzing property of ivermectin was enhanced by the presence of salt, while the efficacy of levamisole and chlorpromazine was reduced. PMID- 12739133 TI - Host specificity and foraging efficiency in blood-sucking parasite: feeding patterns of the flea Parapulex chephrenis on two species of desert rodents. AB - Parasite species can adapt to ecological, behavioral, physiological and biochemical traits of a particular host species. The flea Parapulex chephrenis occurs on the spiny mouse Acomys cahirinus, but does not occur on a co-existing gerbil, Gerbillus dasyurus. To test the hypothesis that the host species affects feeding parameters of a host-specific flea, we studied the feeding rate, rate of blood digestion and resistance to starvation of P. chephrenis when feeding on A. cahirinus and G. dasyurus. We predicted that P. chephrenis would: (1) fill its gut with blood faster, (2) digest blood for a shorter time, and (3) survive longer when starved while feeding on its specific host, A. cahirinus, than on a non-specific host, G. dasyurus. These three responses were observed when P. chephrenis fed on the different hosts and, consequently, our predictions were supported. Twenty percent of fleas filled their midgut after feeding for 10 min on A. cahirinus but this occurred only after 25 min on G. dasyurus. The middle stage of blood digestion was significantly shorter in all fleas feeding on A. cahirinus than in fleas feeding on G. dasyurus. Flea survival was shorter when feeding on G. dasyurus than when feeding on A. cahirinus at 25 degrees C, but no difference in survival time was found at 15 or 20 degrees C. Both A. cahirinus, the specific host, and G. dasyurus, the non-specific host, co-exist in rocky habitats, yet P. chephrenis occurs on one rodent and not the other. The absence of P. chephrenis on G. dasyurus in nature and the decreased foraging efficiency when feeding on this species in the laboratory suggests that some physiological and biochemical differences between hosts can lead to sharp ecological differences in host-parasite relationships. PMID- 12739134 TI - The histone H4 gene of Plasmodium falciparum is developmentally transcribed in asexual parasites. AB - Histones are abundant nuclear core proteins that are present in all eukararyotes and are responsible for linking chromosomes and packaging them into tight chromatin aggregates. The histone H2A, H2B, and H3 genes and a partial sequence of the histone H4 gene from Plasmodium falciparum have been previously identified and share a high level of nucleotide sequence identity. In this study, we compare the histone H4 sequence of the human malaria P. falciparum with the sequences of two mouse malarias, Plasmodium berghei and Plasmodium yoelii, revealing at least 91% identity at the nucleotide level and 100% conservation at the amino acid level. Furthermore, we show the P. falciparum histone H4 is developmentally transcribed in late stage asexual parasites, completing the transcription profile for the genes comprising the histone octamer of P. falciparum and adding support to suggestions that a novel histone mRNA control mechanism exists in this parasite. PMID- 12739135 TI - Paraneoplastic syndromes in ganglioneuroblastoma: contrasting symptoms of constipation and diarrhoea. AB - A paraneoplastic syndrome is occasionally the first clinical symptom seen with tumours. We report on two children who initially presented with paraneoplastic syndromes due to ganglioneuroblastomas: the first with severe watery diarrhoea caused by a ganglioneuroma producing vasoactive intestinal peptide, the second with non-treatable constipation, caused by ganglioneuroma-produced anti-neuronal nuclear antibodies. CONCLUSION: Either severe diarrhoea or chronic constipation may represent rare paraneoplastic syndromes in ganglioneuroblastomas. PMID- 12739136 TI - Tidal off-line exhaled nitric oxide measurements in a pre-school population. AB - Exhaled nitric oxide (ENO) is used as a non-invasive marker of airway inflammation. The aim of this study was to measure ENO in a pre-school population using a relatively novel method, the off-line tidal breathing method, and to investigate differences in ENO between subjects with different presentations of wheezing. ENO was measured in 129 children (median age 4.4 years, quartiles 4.0 4.8 years) through a mouth mask attached to a two-way valve with an expiratory resistance of 5 cm H(2)0. Mean tidal ENO concentration (tENO) was calculated from triplicate measurements. Mean +/- SEM tENO for 89 control subjects was 13+/-0.4 ppb (95%CI 11.8-13.7 ppb); this level was significantly different from tENO in 15 children with a history of recurrent wheezing (18.6+/-1.9 ppb; 95%CI 14.5-22.7 ppb; t-test P<0.0001). Mean tENO in 16 children with a single wheezing episode was 11.4+/-1.0 ppb (95%CI 9.2-13.6 ppb) and thus significantly different from the recurrent wheezing group (t-test P=0.0024). CONCLUSION: The off-line tidal breathing method is a feasible and appealing method for measuring exhaled nitric oxide in pre-school children. With this method, higher tidal exhaled nitric oxide levels were found in children with recurrent wheezing. PMID- 12739137 TI - Three cases of Kingella kingae infection in young children. PMID- 12739138 TI - Effectiveness of rescue antiretroviral therapy including intravenously administered zidovudine and foscarnet in a child with HIV-1 enteropathy. PMID- 12739139 TI - Noonan syndrome with leukaemoid reaction and overproduction of catecholamines: a case report. PMID- 12739140 TI - Functional conservation of the sex-lethal sex determining promoter, Sxl-Pe, in Drosophila virilis. AB - The primary sex determination signal in Drosophila melanogaster, the ratio of X chromosomes to autosomes, sets the activity state of the switch gene, Sex-lethal ( Sxl), by regulating the establishment promoter, m-Sxl-Pe. We have identified and characterized the establishment promoter, v-Sxl-Pe, of the distantly related species Drosophila virilis. Like melanogaster, the virilis Sxl-Pe is organized into four sub-domains: the Sxl-Pe mRNA leader and exon E1 of Sxl protein, the core promoter, the sex-specific element and the augmentation element. The core promoter and sex-specific element of v-Sxl-Pe show considerable sequence similarity to m-Sxl-Pe and contain target sites for components of the X/A signaling system. While the augmentation element of v-Sxl-Pe also has sequence motifs that could function as target sites for the X/A signaling system, it shows little similarity to the melanogaster augmentation element. Functional studies reveal that v-Sxl-Pe drives sex-specific expression in D. melanogaster embryos and that the activity of the virilis promoter is controlled by known components of the melanogaster X/A counting system. Although v-Sxl-Pe responds appropriately to the melanogaster sex determination signal, it is less active than Sxl-Pe from melanogaster. Unexpectedly, the reduced activity is due to differences in the activity of the conserved core promoter, while the non-conserved augmentation element functions effectively. These findings suggest that low-affinity target sites for the X/A counting system are critical for the functioning of Sxl-Pe. PMID- 12739141 TI - A genomewide survey of developmentally relevant genes in Ciona intestinalis. V. Genes for receptor tyrosine kinase pathway and Notch signaling pathway. AB - In the present survey, we identified most of the genes involved in the receptor tyrosine kinase (RTK), mitogen activated protein kinase (MAPK) and Notch signaling pathways in the draft genome sequence of Ciona intestinalis, a basal chordate. Compared to vertebrates, most of the genes found in the Ciona genome had fewer paralogues, although several genes including ephrin, Eph and fringe appeared to have multiplied or duplicated independently in the ascidian genome. In contrast, some genes including kit/flt, PDGF and Trk receptor tyrosine kinases were not found in the present survey, suggesting that these genes are innovations in the vertebrate lineage or lost in the ascidian lineage. The gene set identified in the present analysis provides an insight into genes for the RTK, MAPK and Notch signaling pathways in the ancient chordate genome and thereby how chordates evolved these signaling pathway. PMID- 12739142 TI - A genomewide survey of developmentally relevant genes in Ciona intestinalis. VI. Genes for Wnt, TGFbeta, Hedgehog and JAK/STAT signaling pathways. AB - Cell-cell interactions play important roles in a variety of developmental processes, and therefore molecules involved in the signaling pathways have been studied extensively. Recently, the draft genome sequence of the basal chordate, Ciona intestinalis, was determined. Here we annotated genes for the signaling pathways of Wnt, transforming growth factor beta (TGFbeta), Hedgehog, and JAK/STAT in the genome of Ciona intestinalis. The Ciona genome contains ten wnt genes, six frizzled genes, four sFRP genes, ten TGFbeta family member genes, five TGFbeta-receptor genes, and five Smad genes; most of the genes were found with less redundancy than in vertebrate genomes. The other genes in the signaling pathways are present as a single copy in the Ciona genome. In addition, all of the identified genes for the signaling pathway, except for a few genes, have EST evidence, and their cDNAs are available from the Ciona intestinalis gene collection. Therefore, Ciona intestinalis may provide an experimental system for exploring the basic genetic cascade associated with the signaling pathways in chordates. PMID- 12739144 TI - What is primed in priming from imagery? AB - Priming from imagery is typically weaker than that from perception. This has been interpreted as resulting from weaker activation of perceptual processes. However, for imagery and perception, commonality is only half the story: Each is also characterized by specific processes. If priming can be due to both unshared and shared components of imagery and perception, then it should be possible to observe greater priming from imagery than from perception. Two new priming experiments were designed to test this hypothesis, while controlling incidental task differences. In both experiments, participants studied objects by counting their parts (from a mental image or a picture). Experiment 1 used a word-picture matching test task, which was hypothesized to depend on stimulus processing specific to perception, and Experiment 2 a size judgment test task, which was hypothesized to depend on retrieval and generation processes specific to imagery. As predicted, priming for perceived objects was greater than priming for imagined objects in the word-picture matching task. Conversely, in the size judgment task, more priming from imagery than from perception was observed. These results support the conclusions that (a) imagery and perception have substantial unshared processes, and (b) these processes contribute to priming. PMID- 12739143 TI - Cloning and expression patterns of dystroglycan during the early development of Xenopus laevis. AB - Dystroglycan is a cell surface receptor involved in the pathogenesis of muscular dystrophy, and plays a critical role in the assembly and homeostasis of basement membranes. Since data about the amphibian homologue are limited, we have cloned the full-length dystroglycan cDNAs from the frog Xenopus laevis. Using in situ hybridization, we show that mRNA expression is dynamic, particularly in the notochord at the end of gastrulation and during neurulation, suggesting that the protein might play unexplored roles in the specification and/or formation of this tissue. Subsequently, the transcripts are detected in the otic vesicle, the developing brain, and in mesenchymal cells of the visceral arches, as well as in pharyngeal endoderm, the pronephros, pronephric ducts, proctodaeum and the somites. PMID- 12739145 TI - Determining subprocesses of visual feature search with reaction time models. AB - After the classic serial/parallel dichotomy of visual search mechanisms has been increasingly doubted, we investigated what search mechanisms are used between the two poles termed "pop-out" and "strictly serial search" in an overt feature search paradigm. Since reaction time slopes do not contain sufficient information for this purpose, we developed a novel technique for analyzing reaction times. Individual reaction times are modeled as sums of the durations of successive search steps. Model parameters are task characteristics (similarity, number and arrangement of target and distractors) and processing characteristics of the participant (e.g., attention dwell and shift durations). In Experiment 1, several model variants were fitted numerically to empirical reaction times. The best fitting model suggested that more than one item can be processed in a single fixation, movement of attention is abrupt and not continuous, and even in pop out search, attention is often explicitly moved to the target. In Experiment 2, we measured the central model parameter, the so-called range of attention, more directly and thereby validated the model. The model provides an explanation for the strong variation in the slope of reaction time functions, which is not based on an explicit distinction between parallel and serial search processes. PMID- 12739146 TI - Processing modes and parallel processors in producing familiar keying sequences. AB - Recent theorizing indicates that the acquisition of movement sequence skill involves the development of several independent sequence representations at the same time. To examine this for the discrete sequence production task, participants in Experiment 1 produced a highly practiced sequence of six key presses in two conditions that allowed little preparation so that interkey intervals were slowed. Analyses of the distributions of moderately slowed interkey intervals indicated that this slowing was caused by the occasional use of two slower processing modes, that probably rely on independent sequence representations, and by reduced parallel processing in the fastest processing mode. Experiment 2 addressed the role of intention for the fast production of familiar keying sequences. It showed that the participants, who were not aware they were executing familiar sequences in a somewhat different task, had no benefits of prior practice. This suggests that the mechanisms underlying sequencing skills are not automatically activated by mere execution of familiar sequences, and that some form of top-down, intentional control remains necessary. PMID- 12739147 TI - Lateralized effects of orthographical irregularity and auditory memory load on the kinematics of transcription typewriting. AB - This study investigated the combined effects of orthographical irregularity and auditory memory load on the kinematics of finger movements in a transcription typewriting task. Eight right-handed touch-typists were asked to type 80 strings of ten seven-letter words. In half the trials an irregularly spelt target word elicited a specific key press sequence of either the left or right index finger. In the other trials regularly spelt target words elicited the same key press sequence. An auditory memory load was added in half the trials by asking participants to remember the pitch of a tone during task performance. Orthographical irregularity was expected to slow down performance. Auditory memory load, viewed as a low level stressor, was expected to affect performance only when orthographically irregular words needed to be typed. The hypotheses were confirmed. Additional analysis showed differential effects on the left and right hand, possibly related to verbal-manual interference and hand dominance. The results are discussed in relation to relevant findings of recent neuroimaging studies. PMID- 12739148 TI - Naive impetus and Michotte's "tool effect": evidence from representational momentum. AB - Displacement in the remembered position of targets in displays based on Michotte's (1951/1991) tool effect paradigm was examined. Targets in tool effect displays exhibited less forward displacement than did otherwise identical targets presented in isolation; the decrease in forward displacement was not dependent upon the motion of a visible intermediary, but was dependent upon a visible intermediary contacting both the launcher and the target. The data were consistent with naive impetus theory and the hypothesis that decreases in forward displacement of targets in tool effect displays resulted from the intermediary transferring perceived impetus of the launcher to the target and a dissipation of that impetus with subsequent target motion. Possible connections between displacement, impetus, and the perception of causality are discussed. PMID- 12739149 TI - Evidence for the involvement of cell wall peroxidase in the generation of hydroxyl radicals mediating extension growth. AB - Hydroxyl radicals (*OH), produced in the cell wall, are capable of cleaving wall polymers and can thus mediate cell wall loosening and extension growth. It has recently been proposed that the biochemical mechanism responsible for *OH generation in the cell walls of growing plant organs represents an enzymatic reaction catalyzed by apoplastic peroxidase (POD). This hypothesis was investigated by supplying cell walls of maize ( Zea mays L.) coleoptiles and sunflower ( Helianthus annuus L.) hypocotyls with external NADH, an artificial substrate known to cause *OH generation by POD in vitro. The effects of NADH on wall loosening, growth, and *OH production in vivo were determined. NADH mediates cell wall extension in vitro and in vivo in an H2O2-dependent reaction that shows the characteristic features of POD. NADH-mediated production of *OH in vivo was demonstrated in maize coleoptiles using electron paramagnetic resonance spectroscopy in combination with a specific spin-trapping reaction. Kinetic properties and inhibitor/activator sensitivities of the *OH-producing reaction in the cell walls of coleoptiles resembled the properties of horseradish POD. Apoplastic consumption of external NADH by living coleoptiles can be traced back to the superimposed action of two enzymatic reactions, a KCN-sensitive reaction mediated by POD operating in the *OH-forming mode, and a KCN-insensitive reaction with the kinetic properties of a superoxide-producing plasma-membrane NADH oxidase the activity of which can be promoted by auxin. Under natural conditions, i.e. in the absence of external NADH, this enzyme may provide superoxide (O2*-) (and H2O2 utilized by POD for) *OH production in the cell wall. PMID- 12739150 TI - Molecular and biochemical characterization of an aminoalcoholphosphotransferase (AAPT1) from Brassica napus: effects of low temperature and abscisic acid treatments on AAPT expression in Arabidopsis plants and effects of over expression of BnAAPT1 in transgenic Arabidopsis. AB - Aminoalcoholphosphotransferases (AAPT, EC 2.7.8.1 and EC 2.7.8.2) catalyze the transfer of CDP-aminoalcohols to sn-1, 2 diacylglycerol (DAG) to form phosphatidylaminoalcohols with the release of CMP. The Brassica napus L. AAPT1 gene (designated BnAAPT1) was identified from cDNA libraries of seedlings and developing seeds. Functional characterization was accomplished by heterologous expression of BnAAPT1 in a yeast strain deficient in AAPT activities. BnAAPT1 exhibited a greater preference for utilizing CDP-choline as a substrate with Vmax of 35 [14C]phosphatidylcholine nmol h(-1) mg(-1) protein and apparent Km of 32 microM while CDP-ethanolamine had a Vmax of 13 [14C]phosphatidylethanolamine nmol h(-1) mg(-1)protein and an apparent Km of 127 microM. The enzyme was activated by Mg2+, Mn2+ and phospholipid mixtures, and inhibited by Ca2+. A CDP-alcohol phosphotransferase motif, Asp99-Gly100-(X2)-Ala103-Arg104-(X8)-Gly113-(X3)-Asp117 (X3)-Asp121, was completely conserved in BnAAPT1 and its catalytic role was confirmed by scanning alanine mutagenesis. Over-expression of BnAAPT1 under the control of the double 35S promoter in transgenic Arabidopsis thaliana (L.) Heynh. plants led to elevated levels of the corresponding transcript and enzyme activity. In four of the high over-expression transgenic lines, phospholipid and fatty acid composition analyses revealed that chloroplastidic and extrachloroplastidic membranes isolated from transgenic leaves had about a 25% increase in phosphatidylcholine and in the proportions of polyunsaturated fatty acids [18:2+18:3], relative to the control. There were also consistent, but small differences observed in the proportions of 18:3 in transgenic green siliques and in 20:1 in mature transgenic seeds of these lines. Induction of Arabidopsis AAPT transcription in response to (+)-abscisic acid and low-temperature treatments, and the cold tolerance in BnAAPT1 transgenic seedlings implies that AAPT may play a role in resistance to damage at low growth temperatures. PMID- 12739151 TI - PMR1/SPCA Ca2+ pumps and the role of the Golgi apparatus as a Ca2+ store. AB - Besides the well-known sarco/endoplasmic-reticulum Ca(2+)-transport ATPases (SERCA), animal cells contain a much less characterized P-type Ca(2+)-transport ATPase: the PMR1/SPCA Ca(2+)/Mn(2+)-transport ATPase. SPCA is mainly targeted to the Golgi apparatus. Phylogenetic analysis indicates that it might be more closely related to a putative ancestral Ca(2+) pump than SERCA. SPCA supplies the Golgi apparatus, and possibly other more distal compartments of the secretory pathway, with the Ca(2+) and Mn(2+) necessary for the production and processing of secretory proteins. In the lactating mammary gland, SPCA appears to be the primary pump responsible for supplementing the milk with high (60-100 mM) Ca(2+). It could also play a role in detoxification of cells overloaded with Mn(2+). Mutations in the human gene encoding the SPCA pump ( ATP2C1) result in Hailey Hailey disease, a keratinocyte disorder characterized by incomplete cell adhesion. Recent observations show that the Golgi apparatus can function as a Ca(2+) store, which can be involved in setting up cytosolic Ca(2+) oscillations. PMID- 12739152 TI - Beat-to-beat blood pressure variability and patent ductus arteriosus in ventilated, premature infants. AB - The aim of the present study was to test the hypothesis that a relationship exists between respiratory-induced blood pressure variability (BPV) and transductal shunting in premature infants with respiratory distress. Ten premature infants (27-32 weeks gestation) with respiratory distress ventilated in the synchronised, positive-pressure mode were examined. The interrelations between blood pressure and transthoracic impedance were described using time and frequency domain analysis. Haemodynamic effects of left-to-right transductal shunting were assessed using Doppler echocardiography (ratio of diastolic flow to systolic flow in the subdiaphragmatic aorta). The dependence of blood pressure fluctuations on the respiratory cycle was seen consistently in both time-domain and cross-spectral analysis. The amplitude of these fluctuations varied between infants. In the time domain, the beat-to-beat pressure difference was 0.69-4.1 mmHg for diastolic and 0.99-5.24 mmHg for systolic blood pressure. There was a positive linear relationship between the respiratory-related BPV indicators and the extent of left-to-right transductal shunting ( r=0.86 for diastolic and 0.80 for systolic pressure, P<0.01). Respiratory-related BPV was not correlated to the indicators of left ventricle preload. It is concluded that respiratory related BPV involves both diastolic and systolic blood pressure and is correlated to the magnitude of left-to-right transductal shunting in the population studied. PMID- 12739153 TI - Ca2+ regulatory systems in rat myocardium are altered by 24 weeks treadmill training. AB - The present study was conducted to investigate the effects of long-term exercise training on the main components involved in excitation-contraction coupling and relaxation in rat myocardium. Twenty male Wistar rats were divided into sedentary (S) and treadmill-trained (T) groups. Group T was trained for 24 weeks, 5 days/week (25 m/min, 45-60 min, 0% slope). 48 h after the last exercise session, animals were killed and ventricular and soleus muscle homogenates were obtained. The citrate synthase activity in soleus muscle was significantly increased (163%) in T compared with S rats ( P<0.01), confirming the exercise training efficacy. Although heart weight and cardiac oxidative capacity were not modified by exercise training, the binding of [(3)H] ryanodine and the dihydropyridine [(3)H]PN200-110 to cardiac homogenates, and sarcoplasmic reticulum Ca(2+)-ATPase activity were increased significantly in the ventricular homogenates from T compared with S animals ( P<0.01). Western blot analysis of ventricular homogenates failed to show significant alterations in dihydropyridine receptor and Ca(2+)-ATPase levels in T animals, but revealed an increase of ryanodine receptor density in this group ( P<0.01). The activity of the ectoenzymes 5' nucleotidase and Mg(2+)-ATPase was not affected by training ( P>0.05). In conclusion, long-term treadmill training induces adaptive changes in some of the components of myocardial rat excitation-contraction coupling and relaxation systems that could contribute to the improvement of cardiac function. PMID- 12739154 TI - Mechanically induced potentials in atrial fibroblasts from rat hearts are sensitive to hypoxia/reoxygenation. AB - Membrane potential changes of atrial fibroblasts in response to mechanical stress have been considered to modulate the rhythmic electrical activity of healthy hearts. Our recent findings suggest that cardiac arrhythmia after infarction is related to enhanced susceptibility of the fibroblasts to physical stretch. In this study, we analysed the effect of hypoxia/reoxygenation, which are major components of tissue ischemia/reperfusion, on the membrane potential of atrial fibroblasts. Intracellular microelectrode recordings were performed together with isometric force measurements on isometrically contracting right atrial tissue preparations from adult rats. Lowering the oxygen tension in the perfusate from 80 kPa to 3.5 kPa reduced active force development and decreased the resting membrane potential of the cardiac fibroblasts from -23+/-5 mV to -5+/-2 mV ( n=35). Application of gadolinium (40 microM) to inhibit non-selective cation channels prevented hypoxia-induced membrane depolarization of the fibroblasts. Reoxygenation of the myocardial tissue resulted in a transient increase of the resting membrane potential to maximally -60+/-8 mV. These findings indicate that transmembrane currents in atrial fibroblasts are sensitive to changes in tissue oxygenation. In conclusion, altered electro-mechanical function of the ischemic heart may possibly involve changes of the membrane potential of the cardiac fibroblasts. PMID- 12739155 TI - Effect of acute hyperglycaemia and diabetes mellitus with and without short-term insulin treatment on myocardial ischaemic late preconditioning in the rabbit heart in vivo. AB - Diabetes mellitus (DM) and the resulting hyperglycaemia may interfere with the cardioprotective effect of ischaemic late preconditioning (LPC). Therefore, we investigated the effect of acute hyperglycaemia (part 1) and the effect of alloxan-induced DM with or without short-term insulin treatment (part 2) on LPC. Rabbits, chronically instrumented with a coronary artery occluder, were subjected to 30 min coronary artery occlusion and 2 h reperfusion (I/R) and infarct size (IS) was assessed. In part 1, four groups were studied. Controls were not treated further. LPC induced by a 5-min period of myocardial ischaemia 24 h before I/R reduced IS from 42+/-14 (controls) to 22+/-8% of the area at risk. Hyperglycaemia (600 mg dl(-1) by dextrose infusion, H(600)) before and during the 30 min ischaemia tended to increase IS (57+/-16%, P=0.14 vs. controls) and blocked cardioprotection by LPC (H(600)+LPC, 59+/-19%, P=1.0 vs. H(600), P=0.0003 vs. LPC). In part 2, LPC reduced infarct size from 43+/-13% (control) to 23+/-10% ( P=0.003). In diabetic animals, IS was 39+/-11%, and cardioprotection by LPC could not be elicited (DM+LPC, 41+/-16%, P=0.02 vs. LPC). Short-term insulin treatment (I, 90 min before I/R, blood glucose <150 mg dl(-1)) did not restore the cardioprotective effects of LPC (DM+I, 42+/-15%; DM+LPC+I, 40+/-10%, P=0.03 vs. LPC). It is concluded that acute hyperglycaemia and DM block the cardioprotection induced by LPC in rabbits and that the cardioprotection is not restored by short term insulin treatment. PMID- 12739157 TI - Role of basolateral membrane conductance in the regulation of transepithelial sodium transport across frog skin. AB - Circuit analyses of the principal cell compartment of frog skin ( Rana temporaria and R. esculenta) were made using microelectrode measurements under short-circuit conditions and with the aid of the Na(+) channel blocker amiloride. Under control conditions, intracellular potential ranged between -65 and -5 mV, and the conductances of the apical and basolateral membranes were related directly to the short-circuit current and inversely to the cellular potential. Blockade of apical Na(+) uptake by amiloride hyperpolarized the cells to nearly the same value, irrespective of the potential under transporting conditions. Under these conditions, basolateral membrane conductance increased greatly, which led to paradoxical reactions of the transepithelial Na(+) transport at lower concentrations of amiloride. The half-maximal inhibitory concentration of amiloride estimated from the response of the apical membrane conductance (99+/-10 nM) was about 5 times lower than the value derived from transepithelial current or conductance in the same tissues. The results are discussed in the context of the importance of the membrane potential for acute control of membrane conductance and transepithelial transport. PMID- 12739156 TI - Slow inhibition of N-type calcium channels with GTP gamma S reflects the basal G protein-GDP turnover rate. AB - The inhibition of N-type Ca channels via a G protein pathway is a rapid mechanism for modulating Ca influx. It has been noted, however, that when G proteins are activated by guanosine 5'- O-(3-thiotriphosphate) (GTPgammaS), the speed of inhibition is greatly reduced, despite the pathway having fewer molecular steps. We explored this anomaly in chick dorsal root ganglion neurons by comparing Ca current inhibition using GTPgammaS with application of the G protein receptor agonist noradrenaline. Noradrenaline caused rapid Ca channel inhibition (tau~5 s), contrasting greatly with the ~70-fold slower rate observed with GTPgammaS. Additionally, the slow rate with GTPgammaS could be accelerated to near agonist induced rates by application of noradrenaline, demonstrating that the inhibition with GTPgammaS was not perfusion limited and that the rate-limiting step was upstream from GTPgammaS binding. Our results suggest that in the absence of noradrenaline, G protein activation by GTPgammaS is impeded by the slow resting turnover of GDP/GTP. The rate at which inhibition develops with GTPgammaS (tau~350 s) is thus a direct and sensitive measure of resting GDP turnover. PMID- 12739158 TI - Essential cysteine residues of the type IIa Na+/Pi cotransporter. AB - The rat renal Na(+)/P(i) cotransporter (NaP(i)-IIa) contains 12 native cysteines. When individually replaced by a serine, none appears essential for proper expression and function. Nevertheless, the formation of one essential cysteine bridge (C5/C6), together with a postulated second bridge, is necessary. To determine the minimum cysteine residues required for functional NaP(i)-IIa, with the goal of generating a Cys-less backbone for structure-function studies, mutants were constructed in which multiple endogenous cysteines were replaced by serines in different combinations. In Xenopus oocytes, most mutants were functional, except those where cysteine pairs C4/C9, C4/C12 or C9/C12 were simultaneously deleted. This suggested that one of these pairs could form the second cysteine bridge essential for expression and/or protein function. Up to eight cysteines could therefore be removed to give a functional Cys-reduced NaP(i)-IIa with activity and kinetics comparable to the wild-type (WT). This construct, like all intermediate mutants and the WT, was insensitive to cysteine modifying methanethiosulfonate (MTS) reagents. Moreover, by introducing a novel cysteine into the Cys-reduced NaP(i)-IIa at a site functionally important in the WT (Ser-460), the loss of transport function reported for mutant S460C, after exposure to MTS reagents, was recapitulated. This confirmed that the MTS reagent site of action was Cys-460 and that modification of native cysteines does not contribute to S460C behavior. PMID- 12739159 TI - Carbohydrate supplementation improves moderate and high-intensity exercise in the heat. AB - The aim of the present study was to clarify the effect of carbohydrate (CHO) supplementation on moderate and high-intensity endurance exercise in the heat. Eight endurance-trained men [maximal oxygen uptake ( VO(2max)) 59.5+/-1.6 ml kg( 1) bw(-1), mean+/-SE] cycled to exhaustion twice at 60% VO(2max) and twice at 73% VO(2max) at an ambient temperature of 35 degrees C. Subjects ingested either a 6.4% maltodextrin solution (CHO) or an artificially flavoured and coloured placebo (PLA). Time to fatigue was significantly greater with CHO in both the 60% and 73% VO(2max) trials (14.5% and 13.5% improvement, respectively). Heart rate and oxygen uptake ( VO(2)) did not differ at any point between PLA and CHO. Hypoglycaemia was not seen in any condition but plasma glucose concentrations tended to be higher at both intensities when CHO was fed. CHO oxidation rates were similar at 60% VO(2max) between CHO and PLA. There were no differences between PLA and CHO in the rate of rise of rectal temperatures ( T(rec)) at either intensity but there was a trend for subjects to fatigue at a high temperature when taking CHO. Ratings of perceived exertion (RPE) tended to be lower throughout both CHO trials; this was significant at 80 min and at fatigue at 60% VO(2max). It is concluded that supplementation with CHO improves exercise performance in the heat at both moderate and high endurance intensities. In the absence of a clear metabolic explanation, a central effect involving an increased tolerance of rising deep body temperature merits further investigation. PMID- 12739160 TI - Ion selectivity of stretch-activated cation currents in mouse ventricular myocytes. AB - Stretch-activated non-selective cation currents ( I(SAC)) constitute a mechanism that can induce cardiac arrhythmias. We studied I(SAC) in mouse ventricular myocytes by stretching part of the cell surface between the patch-pipette and a motor-driven glass stylus. In non-clamped cells, local stretch depolarised and induced after-depolarisations and extrasystoles. In voltage-clamped cells (K(+) currents suppressed) I(SAC) activated by local stretch had a nearly linear voltage dependence and reversed polarity between -12 and 0 mV. Conductance G(SAC) increased with the extent of local stretch. I(SAC) was not a Cl(-) current (insensitivity to replacement of Cl(-) by aspartate(-)). I(SAC) was not a Ca(2+) activated current (insensitivity to 5 mM intracellular BAPTA). G(SAC) was blocked by 5 micro M GdCl(3) or by 75 mM extracellular (e.c.) CaCl(2). Removal of e.c. CaCl(2) increased G(SAC) 2.5-fold, as if G(SAC) were sensitive to Ca(2+) and Gd(3+). Replacement of 150 mM e.c. Na(+) by 150 mM Cs(+), Li(+), tetraethylammonium (TEA(+)) or N-methyl d-glucosamine (NMDG(+)) yielded currents that suggested for the conductance a selectivity G(Cs)> G(Na)> G(Li)> G(TEA)> G(NMDG). I(SAC) was suppressed by cytochalasin D, as if an intact F-actin cytoskeleton were necessary for activation of I(SAC). PMID- 12739161 TI - Regulation of K-Cl cotransport by Syk and Src protein tyrosine kinases in deoxygenated sickle cells. AB - Protein tyrosine kinases (PTK) of the Src family are thought to suppress K-Cl cotransport (KCC) activity via negative regulation of protein phosphatases. However, some PTK inhibitors reduce KCC activity, suggesting opposite regulation by different PTK families. We have reported previously that deoxygenation of sickle cells stimulates KCC and activates Syk (a Syk family PTK), but not Lyn (an Src family PTK). In this study the same results were obtained when PTK activities were measured under the conditions used to measure KCC activity and which prevent any change in intracellular [Mg(2+)]. Methyl-2,5-dihydroxycinnamate (DHC), a PTK inhibitor, was more selective for Syk than Lyn, while staurosporine (ST), a broad specificity protein kinase inhibitor, inhibited Lyn more than Syk. Deoxygenation or 4-amino-5-(4-chlorophenyl)-7-( t-butyl)pyrazolo[3,4- d] pyrimidine (pp2, a specific Src inhibitor) stimulated KCC independently. These effects were not additive and were inhibited by DHC. In contrast, ST-induced KCC activation was resistant to DHC, suggesting a different pathway of activation. Overall, these data indicate that Syk activity is required for KCC activation, either induced by deoxygenation of sickle cells, or mediated by Src inhibition in oxygenated cells, and that Syk and Src PTKs exert opposing and interconnected regulatory effects on the activity of the transporter. PMID- 12739162 TI - Isoprenaline-stimulated differential adrenergic response of K+ channels in skeletal muscle under hypokalaemic conditions. AB - The mechanism underlying the hyperpolarization induced by isoprenaline in mouse lumbrical muscle fibres was studied using cell-attached patch and intracellular membrane potential ( V(m)) recordings. Sarcolemmal inwardly rectifying K(+) channels (K(IR): 45 pS) and Ca(2+)-activated K(+) channels (BK: 181 pS) were identified. Exposure to isoprenaline closed K(IR) channels and increased BK channel activity. This increase was observed as a shift from 50 to -40 mV in the voltage dependence of channel activation. Isoprenaline prevented hysteresis of V(m) when the extracellular [K(+)] fell below 3.8 mM. This hysteresis was due to the properties of the K(IR). The effects of chloride transport and isoprenaline on V(m) did not interact purely competitively, but isoprenaline could prevent the depolarization induced by hyperosmotic media equally as well as bumetanide, which inhibits the Na(+)/K(+)/2Cl(-) cotransporter. In lumbrical muscle this leads to hyperpolarization, but this might vary among muscles. The switch from K(IR) to BK as the component of total K(+) conductance was due to isoprenaline. PMID- 12739163 TI - BK channel openers inhibit migration of human glioma cells. AB - Large-conductance Ca(2+)-activated K(+) channels (BK channels) are highly expressed in human glioma cells. However, less is known about their biological function in these cells. We used the patch-clamp technique to investigate activation properties of BK channels and time-lapse microscopy to evaluate the role of BK channel activation in migration of 1321N1 human glioma cells. In whole cells, internal perfusion with a solution containing 500 nM free Ca(2+) and external application of the BK channel opener phloretin (100 micro M) shifted the activation threshold of BK channel currents toward more negative voltages of about -30 mV, which is close to the resting potential of the cells. The concentration of intracellular Ca(2+) in fura-2-loaded 1321N1 cells was measured to be 235+/-19 nM and was increased to 472+/-25 nM after treatment with phloretin. Phloretin and another BK channel opener NS1619 (100 micro M) reduced the migration velocity by about 50%. A similar reduction was observed following muscarinic stimulation of glioma cells with acetylcholine (100 micro M). The effects of phloretin, NS1619 and acetylcholine on cell migration were completely abolished by co-application of the specific BK channel blockers paxilline (5 micro M) and iberiotoxin (100 nM). The phloretin-induced increase in intracellular Ca(2+) was unaffected by the removal of extracellular Ca(2+) and co application of paxilline. These findings indicate that glioma cell migration was inhibited through BK channel activation, independent of intracellular Ca(2+). PMID- 12739164 TI - Pulmonary 15NO uptake in man. AB - Nitric oxide (NO) is commonly thought to reveal more precise values of pulmonary gas uptake through alveolar-capillary membranes (DL) than the normally used carbon monoxide (CO). Since such measurements are influenced by a significant endogenous NO delivery within human airways, we propose the use of the naturally occurring (15)N-labelled stable nitric oxide isotope (15)NO. It occurs with a relative abundance of 0.37% of the dominating isotope (14)NO. Therefore, the endogenous (15)NO production can be neglected. In the present pilot study we demonstrate the workability of (15)NO in determining DL in healthy individuals. In seven female and 15 male volunteers, averaged values of DL increase with increasing mean alveolar volume as well as individual body height ( P=0.000001). Due to the very high significance level obtained from the multiple regression analysis, we conclude that the application of (15)NO establishes a novel approach to calculate standard values of DL. Such calculations can be employed to predict a reference for patients who suffer from pulmonary diffusion limitation. PMID- 12739165 TI - Human skeletal muscle mitochondrial metabolism in youth and senescence: no signs of functional changes in ATP formation and mitochondrial oxidative capacity. AB - The mitochondrial theory of ageing was tested. Isolated mitochondria from the quadriceps muscle from normal, healthy, young (age 20+ years, n=12) and elderly (70+ years, n=11) humans were studied in respiratory experiments and the data expressed as activities of the muscle. In each group, the subjects exhibited a variation of physical activity but, on average, the groups were representative for their age with maximum O(2) consumption rate of 50+/-9 and 34+/-13 ml min(-1) kg(-1) (mean+/-SD), respectively. Thirteen different activities were assayed. alpha-Glycerophosphate oxidation was lower in the 70+ group (38%, P~0.001), as was the respiratory capacity for fatty acids (19%, P~0.03). The remaining eleven activities, including those of the central bioenergetic reactions, were not lower in the 70+ group. Pyruvate and alpha-ketoglutarate dehydrogenase activities (i.e. the tricarboxylic acid cycle turnover) and the respiratory chain activity could all account for ~14 mmol O(2) min(-1) kg(-1) muscle (37 degrees C). The capacity for aerobic ATP synthesis was ~35 mmol ATP min(-1) kg(-1). The mitochondrial capacities were far in excess of whole-body performance. They were related to physical activity, but not to age. The mitochondrial theory of ageing, which attributes the age-related decline of muscle performance to decreased mitochondrial function, is incompatible with these results. PMID- 12739167 TI - FEPS in Nice: a date with the best of European physiology. PMID- 12739166 TI - Tympanic temperature reflects intracranial temperature changes in humans. AB - The purpose of the study was to identify extracranial locations in which temperature changes in humans reflect those of intracranial temperature in a reliable and repeatable way. This was achieved by subjecting 14 non-anaesthetized patients after neurosurgery to face fanning while intracranial and extracranial temperatures were continuously measured. In all patients the cranium was closed and the group included both febrile and non-febrile as well as hyperthermic and normothermic patients. The patients' faces were fanned for 20-30 min, with a small fan at an air speed of 3.25 m s(-1). This gave intracranial temperature changes measured in the subdural space ( T(sd)) that were highly and significantly correlated ( r=0.91, P<0.05, n=14) with changes in tympanic temperatures ( T(ty)). A low, statistically insignificant correlation ( r=0.40, P>0.05, n=12) was found between T(sd) and oesophageal temperatures. In conclusion, intracranial temperature changes, induced by face fanning, were reliably reflected by the changes in T(ty). PMID- 12739170 TI - The acetyl-CoA transporter family SLC33. AB - The acetyl-CoA (Ac-CoA) transporter (AT-1) is a multiple transmembrane protein in the endoplasmic reticulum. Ac-CoA is transported to the lumen of the Golgi apparatus, where it serves as the substrate of acetyltransferases that modify the sialyl residues of gangliosides and glycoproteins. The AT-1 gene, originally named ACATN (acetyl-CoA transporter), was cloned from human melanoma cells. Although homologs of this family of proteins have been identified in lower organisms, such as Escherichia coli, Drosophila melanogaster, and Caenorhabditis. elegans, currently only one member of this SLC33A1 family has been identified in humans. Thus, SLC33A1 proteins should be re-named ACATN1 or AT-1. Although acetylated gangliosides show a highly tissue-specific distribution, AT-1 is ubiquitously expressed. Phylogenetically, the AT-1 gene is highly conserved, suggesting that it is particularly significant. The precise physiological roles of this transporter protein, however, remain to be elucidated. PMID- 12739169 TI - The SLC16 gene family-from monocarboxylate transporters (MCTs) to aromatic amino acid transporters and beyond. AB - The monocarboxylate cotransporter (MCT) family now comprises 14 members, of which only the first four (MCT1-MCT4) have been demonstrated experimentally to catalyse the proton-linked transport of metabolically important monocarboxylates such as lactate, pyruvate and ketone bodies. SLC16A10 (T-type amino-acid transporter-1, TAT1) is an aromatic amino acid transporter whilst the other members await characterization. MCTs have 12 transmembrane domains (TMDs) with intracellular N- and C-termini and a large intracellular loop between TMDs 6 and 7. MCT1 and MCT4 require a monotopic ancillary protein, CD147, for expression of functional protein at the plasma membrane. Lactic acid transport across the plasma membrane is fundamental for the metabolism of and pH regulation of all cells, removing lactic acid produced by glycolysis and allowing uptake by those cells utilizing it for gluconeogenesis (liver and kidney) or as a respiratory fuel (heart and red muscle). The properties of the different MCT isoforms and their tissue distribution and regulation reflect these roles. PMID- 12739168 TI - Molecular physiology of cation-coupled Cl- cotransport: the SLC12 family. AB - The electroneutral cation-chloride-coupled cotransporter gene family ( SLC12) was identified initially at the molecular level in fish and then in mammals. This nine-member gene family encompasses two major branches, one including two bumetanide-sensitive Na(+)-K(+)-2Cl(-) cotransporters and the thiazide-sensitive Na(+):Cl(-) cotransporter. Two of the genes in this branch ( SLC12A1 and SLC12A3), exhibit kidney-specific expression and function in renal salt reabsorption, whereas the third gene ( SLC12A2) is expressed ubiquitously and plays a key role in epithelial salt secretion and cell volume regulation. The functional characterization of both alternatively-spliced mammalian Na(+)-K(+) 2Cl(-) cotransporter isoforms and orthologs from distantly related species has generated important structure-function data. The second branch includes four genes ( SLC12A4- 7) encoding electroneutral K(+)-Cl(-) cotransporters. The relative expression level of the neuron-specific SLC12A5 and the Na(+)-K(+)-2Cl( ) cotransporter SLC12A2 appears to determine whether neurons respond to GABA with a depolarizing, excitatory response or with a hyperpolarizing, inhibitory response. The four K(+)-Cl(-) cotransporter genes are co-expressed to varying degrees in most tissues, with further roles in cell volume regulation, transepithelial salt transport, hearing, and function of the peripheral nervous system. The transported substrates of the remaining two SLC12 family members, SLC12A8 and SLC12A9, are as yet unknown. Inactivating mutations in three members of the SLC12 gene family result in Mendelian disease; Bartter syndrome type I in the case of SLC12A1, Gitelman syndrome for SLC12A3, and peripheral neuropathy in the case of SLC12A6. In addition, knockout mice for many members of this family have generated important new information regarding their respective physiological roles. PMID- 12739171 TI - In vitro evaluation of the efficacy of skin barrier creams and protective gloves on percutaneous absorption of industrial solvents. AB - OBJECTIVES: The aim of the experiments was to evaluate the efficacy of skin barrier creams (SBCs) and protective gloves and its potential for reduction of percutaneous absorption of industrial solvents. METHODS: We assessed percutaneous absorption of ethylene glycol (EG), isopropyl alcohol (IA) and 1,2,4 trimethylbenzene (TMB), using static diffusion cells. These solvents were applied neat (EG, TMB) as well as in 10% and 50% aqueous solution (EG, IA) or in 10% and 50% ethanol-diluted solution (TMB). Furthermore, we tested the percutaneous absorption of IA mixed in one cleaning agent (CA), used in newspaper printing shops to clean the rollers of printing machines. Additionally, the penetration behaviour of 10% and 50% solutions of EG, IA and TMB was tested. The experiments were carried out on untreated and on SBC-treated excised human skin from one donor, and on protective gloves. Saline was used as receptor fluid for EG and IA, and neat ethanol for TMB. RESULTS: The penetration of 50% EG, IA and TMB solutions through SBC-treated skin was higher than in untreated skin (factor 3.9 for EG, 0.32 for IA and 0.06 for TMB). The penetration of IA in the IA-CA mixture was five-times higher through untreated skin as for the single compound in 10% aqueous solution. In skin, treated with SBC, we found a 17-fold penetration enhancement of IA in the IA-CA mixture. No appreciable penetration of EG and IA was observed through nitrile rubber gloves. CONCLUSIONS: Our in vitro experiments could not demonstrate an efficacy of SBC to protect skin penetration for the tested solvents. The percutaneous absorption of all solvents in 50% solution was increased through skin treated with SBCs. Furthermore, SBCs enhance the penetration rates of solvents from complex mixtures compared with the single solvents. The tested gloves showed sufficient protection for the hydrophilic solvents, but not for TMB. PMID- 12739172 TI - Risk factors for musculoskeletal disorders among nursing personnel in Greek hospitals. AB - OBJECTIVES: To investigate the relationships between physical, psychosocial, and individual characteristics and different endpoints of musculoskeletal complaints of the lower back, neck and shoulders. METHODS: In this cross-sectional study a questionnaire survey was carried out among 351 nursing personnel (response 84%) in six general hospitals in Athens, Greece. A questionnaire was used on physical and psychosocial workload, need for recovery, perceived general health and (1) the occurrence of musculoskeletal complaints in the past 12 months, (2) chronic complaints during at least 3 months, and (3) complaints which led to sickness absence. In logistic regression analysis odds ratios (ORs) were estimated for all relevant risk factors. RESULTS: Self-reported factors of physical load were associated with the occurrence of back pain (OR=1.85), neck pain (OR=1.88), and shoulder pain (OR=1.87) but these factors were not associated with chronic complaints and musculoskeletal sickness absence. Physical load showed a trend with the number of musculoskeletal complaints with ORs of 2.47 and 4.13 for two and three musculoskeletal complaints, respectively. No consistent influence of psychosocial factors on complaints, chronicity, or sickness absence was observed. A perceived moderate general health was also a risk factor, and strongest associations were observed for sickness absence due to back pain (OR=2.03), neck pain (OR=8.31), and shoulder pain (OR=6.84). CONCLUSIONS: The handling of physical loads among nurses seems to put them at risk for the occurrence of musculoskeletal disorders. The development of these complaints into chronic complaints and associated sickness absence is strongly determined by perceived general health and almost not associated with work-related physical and psychosocial risk factors. When the influence of work-related risk factors on musculoskeletal health is being investigated, the general health status of individual workers should be taken into account. PMID- 12739174 TI - Comparison of three different technologies for pupil diameter measurement. AB - BACKGROUND: The pupil diameter plays an important role in the occurrence of photopic phenomena after refractive surgery. Standardized estimation can be performed using a lens system with a built-in millimeter scale (Colvard, Oasis Medical, California). A new computerized technique allows dynamic and binocular measurement of the pupil diameter by use of infrared light (P2000SA, Procyon Instruments, London, UK). An additional approach is a wavefront aberrometer based on the Hartmann-Shack principle (WASCA; Asclepion-Meditec-Zeiss, Jena, Germany). These strategies were compared. DESIGN: Non-randomized comparative trial. PARTICIPANTS AND METHODS: The pupil diameter of 56 eyes of 28 probands (18 female, mean age 23 years) was measured under scotopic conditions by three independent examiners with each measurement device. The measurement devices were compared intraindividually by pairwise sign tests. Description was based on the intraindividual differences' medians and quantiles. RESULTS: Median pupil diameters were 6.67 mm for the scale pupillometer (interquartile range 6.07-6.94 mm), 6.60 mm for the dynamic pupillometer (6.0-7.02 mm), and 6.37 mm for the wavefront-based aberrometer (5.9-6.7 mm). Pairwise comparison revealed statistically significant ( P<0.05), although not clinically relevant median deviations. Although no clinically relevant median differences were observed (when based on intraobserver means), deviations for single pupil diameter assessments ranged up to 1 mm. CONCLUSION: No clinically relevant median deviations were observed in the underlying repeated measurement scenario. The scale pupillometer showed greater interobserver variation than the objective tests. PMID- 12739177 TI - The Center for Human Appearance: a multi-specialty concept. AB - At the time of its founding in 1988, The Edwin and Fannie Gray Hall Center for Human Appearance at the University of Pennsylvania was the first such center in a major academic setting dedicated to the multispecialty serious study of problems of appearance. The concept was for five key specialties, originally, and now six, with primary interests in appearance, to meet regularly to present ideas and problems, and to approach the study and treatment of appearance in a comprehensive, scholarly, and multidisciplinary manner. The group has been cohesive, which has depended on frequent and regular personal interactions, as well as an adequate and supportive financial structure. With these factors in place, and a dedication to making it work, there have been publications, new ideas, and teamwork that has made it possible for competing specialties to effectively function together. PMID- 12739178 TI - Body dysmorphic disorder in cosmetic surgery patients. AB - This article discusses body dysmorphic disorder (BDD) among cosmetic surgery patients. BDD is characterized as a preoccupation with a slight or imagined defect with some aspect of physical appearance that leads to significant disruption in daily functioning. Although the prevalence of BDD within the general population is unknown, recent evidence suggests that the disorder may be overrepresented among persons who seek cosmetic medical treatments. Preliminary evidence suggests that persons with BDD do not benefit from cosmetic treatments and frequently experience a worsening of their BDD symptoms. Thus, identification of BDD symptomotology has become an important part of determining patients' appropriateness for cosmetic procedures. PMID- 12739175 TI - Sub-retinal hemorrhage during internal limiting membrane peeling for a macular hole. AB - PURPOSE: To report the occurrence of sub-retinal hemorrhage during peeling of the internal limiting membrane (ILM). CASE REPORTS: In case 1, a three-port pars plana vitrectomy was performed on a 57-year-old woman with a macular hole. Following the staining of the ILM with indocyanine green, the ILM was peeled with forceps, and a sub-retinal and vitreous hemorrhage resulted from a vein during the peeling. The bleeding was stopped by elevating the intraocular pressure. The final visual acuity was 20/30. In case 2, similar procedures were performed on a 68-year-old woman with a macular hole. A sub-retinal hemorrhage occurred while peeling the ILM. The pre-operative visual acuity was 20/300, and final visual acuity was 20/200. CONCLUSIONS: Surgeons should be aware that sub-retinal and vitreous hemorrhage can be a complication of ILM peeling. PMID- 12739179 TI - Nasal reconstruction: aesthetic and functional considerations for alar defects. AB - The nose is the most common site for cutaneous malignancy, and nasal defects requiring reconstruction frequently result from tumor resection. These defects present both a functional and an aesthetic challenge to the surgeon. The ala is a unique structure that plays a key role in the airway as well as in the harmony of nasal geometry. Because it is a paired structure, the restoration of the premorbid aesthetic and anatomic state of the ala is crucial in achieving an optimal result. This article focuses on alar reconstruction, with discussion of the anatomy and the aesthetic and functional considerations that lead to successful restoration of the ala. An algorithm outlining options for nasal defects involving the ala is be presented. PMID- 12739180 TI - Repair of the unilateral cleft lip/nose deformity. AB - Successful surgical repair of the unilateral cleft lip and nose deformity, defined as normal orbicularis oris function and near-perfect symmetry of the repaired lip and nose, demands that the surgeon possess complete understanding of the embryology and anatomy of the midfacial defects. The surgical approach to repair of the unilateral cleft lip/nose should place great emphasis on achieving symmetry, not only with the lip segments but also perhaps even more importantly with the nasal tip. The reconstruction should recreate an intact fully functional orbicularis oris muscle across the cleft and camouflage the scar optimally. We have found that modification of the Millard rotation-advancement flap technique, with particular attention to the primary nasal repair, provides the best outcomes. In patients who have undergone primary repair of the lip and/or nose deformity, secondary rhinoplasty is generally required, regardless of the technique used at the primary repair. The degree of nasal deformity, however, is less severe following primary repair of the asymmetric nasal tip. We have found that the sliding flap cheliorhinoplasty, Wang's modification of the Vissarionov technique, provides excellent results for most secondary cleft rhinoplasties. PMID- 12739181 TI - Auricular cartilage in revision rhinoplasty. AB - Revision rhinoplasty is a challenge in reconstruction to the rhinoplasty surgeon, both in the techniques of repair and the choice of implant material for augmentation grafting. Often, patients seeking revision or reconstructive rhinoplasty have previously undergone septoplasty with sacrifice of major amounts of septal cartilage. These situations confront the surgeon with the need for a decision about the material that will be used for structural grafting. The senior author follows the time-tested approach of generations of surgeons who have used exclusively autogenous material for nasal reconstruction because of its superior long-term survival characteristics, its ready availability in the head and neck region, its resistance to infection and resorption, and its bendability and flexibility when implanted in the nose. With this in mind, the subject of this article is the use of auricular cartilage in revision rhinoplasty. Careful strategic planning must be undertaken to get the maximal and ideal benefit from the auricular cartilage. The revision rhinoplasty surgeon must understand the anatomy of the external ear and must be able to manage the precious cartilage supply to get the maximum use of it in reconstructive rhinoplasty. PMID- 12739182 TI - Surgery of the auricle. AB - Auricular reconstruction is a unique area of facial plastic surgery where a wide array of reconstructive options often must be considered. The external ear is unique in its aesthetic role where the normal auricle often goes unnoticed; yet even a small irregularity can stand out and become conspicuous. The reconstruction of large or total auricular defects is a combination of science and art. Two forms of auricular reconstruction are discussed: (1) those for a congenitally abnormal shape but no acquired tissue deficiency, that is, otoplasty, and (2) repairs requiring a reconstruction of discrete loss of tissue. A general algorithm is presented that can assist with flap selection and covers the techniques for grafts, framework repair, local and pedicled flaps, temporoparietal facial flaps, and auricular prostheses. PMID- 12739183 TI - Genioplasty. AB - Genioplasty has been a useful and frequently employed technique in the aesthetic facial surgeon's armamentarium. However, as the prevalence of obstructive sleep apnea becomes more apparent and its diagnosis more frequent, genioplasty has also evolved to a commonly performed procedure for reconstruction of the upper airway. This article discusses patient evaluation and surgical techniques used in aesthetic and functional surgery of the chin. Because these topics have been discussed extensively in the literature, this will serve as a synopsis of current techniques. PMID- 12739184 TI - Enhancement of the fibula free flap by alveolar distraction for dental implant restoration: report of a case. AB - We describe the utilization of distraction osteogenesis in the free fibular microvascular bone graft to the mandible for increasing bone height for future osseointegrated dental implants. Successful reconstruction of a resected mandible requires restoration of both function and esthetic form. Although current reconstructive techniques restore anterior-posterior and lateral projection, often the graft's vertical height is not sufficient for the placement of osseointegrated dental implants and subsequent oral prosthesis. The patient was a sixteen-year-old male who was found to have a large desmoplastic fibroma of the left mandible, which was resected. The defect was successfully restored with a free fibular microvascular bone graft and reconstruction plate. Nevertheless, the patient had persistent problems with mastication and it was decided to perform a segmental osteotomy of the neomandible. Two internal vertical distraction devices were then placed in the mandible. The appliances were then activated five days postoperative, twice a day, for a total of 14 days. At that time 1.5 cm of distraction had occurred and the patient was placed in a consolidation phase for four months. The patient then had sufficient bone height and was restored with 8 osseointegrated dental implants. PMID- 12739185 TI - The deep plane facelift. AB - The goal of any cosmetic facial plastic surgery procedure is to obtain a natural appearing result that is not overdone or obviously operated upon. This is the case whether the background of the operating surgeon is in facial plastic and reconstructive surgery, general plastic surgery, dermatology, or oculoplastic surgery. To satisfy this goal, the surgeon must apply those techniques that consistently achieve effective but understated results rather than experimenting with every new innovation or fad. More and more surgeons are moving away from some of the so-called advances of the last decade, including the use of lasers for skin resurfacing and the application of endoscopic approaches to the face, in the search for the best results possible. Sometimes this means utilizing older techniques that have proven the test of time, such as the coronal browlift and chemical peels, whereas other times this means embracing those newer techniques that clearly represent major advances in our specialty, such as the transconjunctival approach to lower lid blepharoplasty. The deep plane facelift is one such advance. In contrast to traditional, skin or short-SMAS flap techniques, the deep plane facelift allows the surgeon to consistently obtain a natural-appearing result that not only addresses the key areas of concern in the aging face but also proves to be longer lasting as well. The deep plane technique also results in fewer complications, an improved ability to address the melolabial region and jowls, and increased anatomic appeal for the operating surgeon. PMID- 12739186 TI - A systematic comprehensive approach to management of irreversible facial paralysis. AB - Irreversible facial palsy (IFP) presents a multitude of problems arising from a paretic periorbital and facial complex, the solutions to which cross the spectrum of multiple specialties. The process of facial rehabilitation can be simplified by subdividing the face into functional units. These units consist of the brow complex, the periorbital complex, the midface complex, and the lower face/oral complex. Although all of these units are interrelated and influence each other, careful study of the deformity and symptoms of each unit yields a coherent approach and customized surgical plan. The following provides a complete evaluation method for the surgeon to review and customize an approach to the individual patient's needs and desires. Facial rehabilitation must be tailored to each individual, addressing both functional as well as aesthetic concerns for each facial unit. PMID- 12739187 TI - Management of adult facial vascular anomalies. AB - Vascular anomalies are best classified by biologic behavior-active hemangiomas, and inactive vascular malformations. Hemangioma growth is restricted to infancy, but residual deformity may persist to adult life in the form of redundant skin, bulky fibrofatty tissue, and ectatic cutaneous vessels. Treatment largely consists of scar revision and laser photocoagulation of residual vascular pigmentation. Vascular malformations are present throughout life and can cause increasing facial distortion and disfigurement as capillaries, veins, lymphatics, and/or arteries further dilate and the facial features hypertrophy. A variety of treatment options include laser photocoagulation, sclerotherapy, embolization, and direct surgical excision, and often a combination of techniques yields improved results. In many cases, however, therapy is only palliative, and patients should be advised of the limitations of each treatment option to avoid unrealistic expectations and disappointing results. PMID- 12739188 TI - Laser hair removal. AB - Laser-assisted hair removal has rapidly emerged as a noninvasive technique for long-term reduction in unwanted hair. This article reviews the pertinent aspects of hair follicle biology, the theoretical aspects of the physics and mechanisms of laser epilation, the different types of lasers available for hair removal, and the practical clinical aspects of laser hair removal. PMID- 12739189 TI - Facial fat grafting: the search for predictable results. AB - Since the majority of volume loss to the face is due to fat atrophy, autologous fat grafting with living fat cells represents the ideal tissue replacement. Recent technical improvements have led to excellent results utilizing fat grafting for cosmetic and reconstructive indications. The purpose of this article is to describe our clinical and laboratory experience with successful facial fat grafting for volume augmentation. Our laboratory experience with facial fat grafting centers around the results of multiple studies utilizing a nude mouse model of facial fat transplantation. Armed with the knowledge gained from our laboratory experience we present modified techniques to optimize fat grafting in the clinical setting. Furthermore, we present the results of several clinical studies examining a variety of recipient sites including nasolabial folds, glabella, lips, and lower eyelids. Lastly we describe our experience utilizing fat grafting to treat patients with hemifacial atrophy. When utilized in the appropriate areas, facial fat grafting can provide long-lasting aesthetically superior replacement for the soft tissues lost through aging or disease. PMID- 12739192 TI - Becoming the Employer of Choice. PMID- 12739193 TI - Of hubris and hope: transforming nursing for a new age. PMID- 12739194 TI - Minimum staffing ratios: the California workforce initiative survey. PMID- 12739195 TI - Magnet and Baldrige: SSM Health Care's journey for excellence (Part I of II). PMID- 12739196 TI - Pieces of the shortage puzzle: aging and shift work. AB - Addressing issues important to the aging nurse and night-shift worker is imperative for retention success. It is important for nurse leaders to create an environment that will keep older nurses and night-shift workers working. The good news is that health care leaders are talking. Research is being done and will hopefully continue. Now it is time for health care leaders to welcome nurses, educators, and policymakers into their circle of discussion to preserve quality health care and retain our nurses at the bedside. PMID- 12739197 TI - Analyzing and promoting issues in health policy: nurse manager's perspective. PMID- 12739198 TI - The advantages of keeping the staff well. PMID- 12739199 TI - Connected: communication skills for nurses using the electronic medical record. PMID- 12739200 TI - Implementing clinical IT in critical care: keys to success. AB - Ensuring the success of a clinical information system in critical care requires careful selection of the right system to address the unique needs in this area. In addition, the methodology used for implementation must include key stakeholders, ensure nursing and physician leadership, understand and improve clinical processes, and provide ongoing training and support. These guidelines can be applied to the implementation of any clinical information system. They provide the opportunity to demonstrate value and benefits from CIS in critical care and beyond. PMID- 12739201 TI - On leadership organizational intelligence/organizational stupidity: the leader's challenge. AB - Creating organizations with a high IQ or creating organizations without the necessary intelligence guarantees success or failure of the organization. Without structures such as shared leadership and other forms of participative management, the organization or unit cannot access and use the available information and wisdom in the organization. When nurses and other health care professionals do not feel like they have a shared stake and do not feel like citizens of the organization, they lack passion for the organization's work. When nurses feel a sense of share ownnership and autonomy for the clinical practice, terrific outcomes are achieved. Leaders must accept the challenge to build the infrastructure that leads to excellence in organizational IQ. PMID- 12739202 TI - Untangling the lines: using a transfer center to assist with interfacility transfers. PMID- 12739203 TI - Vision, structure, and resolve. PMID- 12739204 TI - Rocket science. PMID- 12739205 TI - Transformations of 21st century health care. Part 2. That giant sucking sound. AB - Technology is a powerful force that is drawing health care away from secondary care hospitals. Those who adapt best to new technology will thrive in the coming decades. PMID- 12739206 TI - Share the knowledge. AB - Consumers are demanding trust, integrity and honesty from health care leaders and providers. CEOs and physicians need new ways to work with their colleagues and patients. PMID- 12739207 TI - IDEOlogy. AB - A group of iconoclastic designers has taught the art of innovation to some major hospital systems. Their five-step formula elicits breathtaking changes for patients and staff. PMID- 12739209 TI - Small steps toward integrating CAM. PMID- 12739208 TI - Time for reconciliation. PMID- 12739210 TI - Building better performance. PMID- 12739211 TI - The end of the community hospital. PMID- 12739212 TI - Financing the future. PMID- 12739213 TI - Look up. PMID- 12739214 TI - Capital crunch. PMID- 12739215 TI - Need for cash. PMID- 12739216 TI - Cancer advice online. PMID- 12739217 TI - Planet health. PMID- 12739218 TI - A flawed plan. PMID- 12739219 TI - Rights of passage. PMID- 12739220 TI - Into the fold. PMID- 12739221 TI - Spotting the difference. PMID- 12739222 TI - Safe at work. PMID- 12739223 TI - Watch out for measles. PMID- 12739224 TI - Attitudes towards a high security hospital. AB - AIM: To study attitudes towards Rampton Hospital by reviewing the attitudes of external students and professionals in training following an arranged guided tour of the hospital; and examining media coverage of the hospital over one year. METHOD: All visitors had a guided day tour of Rampton. Everyone was given a questionnaire to complete at the end of the tour, and responses were reviewed. Rampton public relations department collects all print media coverage and monitors broadcasts in the UK. The department then assimilates this information into an annual report, which was examined for the relevant year. RESULTS: Questionnaires were given to 995 visitors and completed by 416 (42 per cent). Of those that returned the questionnaires, 95 per cent (n = 395) rated their satisfaction with the tour as either 'satisfied' or 'very satisfied'. A positive change in attitude was indicated by approximately one quarter of respondents (n = 105) following their tour. There were no negative changes in attitude expressed. Of the 203 press cuttings relating specifically to the hospital, 59 (29 per cent) were classed as positively enhancing and building the reputation of the hospital, 86 (42 per cent) were classed as negative and 58 (29 per cent) were classed as neutral. CONCLUSION: The tour of Rampton was valuable in bringing about a positive attitude change in the visitors. Although some media coverage assessed was negative, it had been expected to be more so. The authors wonder whether people's negative impressions are formed from the wider media, such as television dramas and movies. PMID- 12739225 TI - Promoting excellence: a draft RCN position statement. AB - The purpose of this draft position statement on nursing research and development is to serve as a consultation document, in the profession and with other key stakeholders, throughout April and May 2003. Thereafter, a formal position statement will be published. PMID- 12739226 TI - Management of autonomic dysreflexia. AB - Autonomic dysreflexia is a syndrome that affects those with a spinal cord lesion above the mid-thoracic level. The symptoms of the syndrome, its causes, how to recognise it, and how to alleviate it are described. The author explains how patients can be educated to recognise the symptoms of the condition, and take action to alleviate them. PMID- 12739228 TI - A different interpretation. PMID- 12739227 TI - Insulin resistance. AB - Insulin resistance is one of the most important causes of premature death in developed countries (Turtle 2000). In this article, Alison Jeffery examines the basic pathology of insulin resistance and its effect on metabolic health. The role of the nurse is discussed in relation to prevention, health promotion and drug treatments for the management of this condition. PMID- 12739229 TI - Stereolithography: neurosurgical and medical implications. AB - We present material to define and understand the concept of Stereolithography (STL) and its potential benefits to the field of neurosurgery and other medical specialties. A historical and scientific review of the literature on stereolithography, its evolution and uses in neurosurgery, forensic medicine, and other medical specialties are described. Considerations regarding different techniques used to obtain STL are discussed. The reproduction of cranial and vascular structures using this technique is evaluated. Data acquisition and model fabrication are the two basic steps required for stereolithography to create custom models for multiple applications in cranio-facial surgery, vascular studies, orthopedic surgery, urology and forensic medicine, among others. Stereolithography is a relatively new technique which continues to grow in many medical fields. Pre-operative education of patients, better understanding of patient anatomy, and the creation of custom-made prostheses are proven benefits of this technique. PMID- 12739230 TI - Endolymphatic sac tumor: staged endovascular-neurosurgical approach. AB - Endolymphatic sac tumors are rare, low-grade adenocarcinomas of the petrous bone. These tumors are highly vascularized and surgical removal is often challenging. We present a staged endovascular-neurosurgical approach consisting of stent assisted angioplasty of the petrous carotid, selective endovascular embolization and surgical removal. PMID- 12739231 TI - Relationship between expression of O6-methylguanine-DNA methyltransferase, glutathione-S-transferase pi in glioblastoma and the survival of the patients treated with nimustine hydrochloride: an immunohistochemical analysis. AB - Drug resistance is one of the important factors that determine tumor response to chemotherapy. Several candidates for resistance to various chemotherapeutic agents have been elucidated. O6-methylguanine-DNA methyltransferase (MGMT) removes methylation damage induced by nitrosourea from the O6 position of DNA guanines before cell injury. Glutathione-S-transferase (GST) pi is also involved in nitrosourea resistance. We examined the expression of MGMT and GST pi in 18 glioblastomas (GBM) using immunohistochemistry and compared the results with patients' survival after administration of 1-(4-amino-2-methyl-5-pyrimidinyl) methyl-3-(2-chloroethyl)-3-nitrosourea hydrochloride (ACNU)-based chemotherapy. According to the Kaplan-Meier's method, although median progression free survival (PFS) of eight patients whose tumors retained high MGMT (3+ approximately 2+), and 10 patients whose tumors showed low MGMT expression (1+ approximately 0) were nine and 15 months, respectively (p = 0.09), median overall survival (OS) of the two groups were 12 and 22 months, respectively, which were significantly different (p = 0.01). GST pi expression in GBM was not a prognostic factor. It is suggested that GBM with strong staining of MGMT activity may show more resistance to ACNU-based chemotherapy compared to that with low MGMT. The simple immunohistochemical analysis of MGMT in GBM can be a useful method to determine whether ACNU or another treatment regimen should be recommended. PMID- 12739232 TI - Protection against ischemic brain damage by GDNF affecting cell survival and death signals. AB - Neuroprotective effects of glial cell line-derived neurotrophic factor (GDNF) on cell survival and death signals were investigated after 90 min of transient middle cerebral artery occlusion (MCAO) in rats. Immunoreactivities of phosphorylated Akt (p-Akt), cleaved caspase-9 (c-cas9), and -3 (c-cas3) increased after the reperfusion in the penumbra in vehicle group with peaks at 3 h, 8 h, and 1 day, respectively. Topical application of GDNF (6.8 micrograms/9 microliters) on brain surface potentiated and prolonged p-Akt activation, but suppressed activation of the caspases, and reduced the number of terminal deoxynucleotidyl transferase-mediated dUDP-biotin in situ nick labeling (TUNEL) positive cells. These results suggest that GDNF plays a protective role against ischemic injury by controlling the balance between Akt pathway and caspase cascades. PMID- 12739233 TI - The long-term effect of recombinant tissue-plasminogen-activator (rt-PA) on edema formation in a large-animal model of intracerebral hemorrhage. AB - Hematoma puncture, fibrinolysis, and aspiration of the liquefied clot is a promising new treatment strategy for large intracerebral hemorrhages (ICH). Characteristics of the cellular injury and neuronal and glial cell death associated with ICH and the administration of fibrinolytic agents still need to be defined. We developed a porcine model to study the histopathological effects of recombinant tissue-Plasminogen-Activator (rt-PA) on perihematomatous cell integrity. In 20 pigs, lobar hematomas were induced by intracranial pressure (ICP)-controlled injections of 7.6 +/- 1.6 ml of autologous blood into the white matter of the right frontal hemisphere. In nine animals, the clots were lysed with rt-PA, thereby facilitating aspiration 2 h after hematoma induction. In 11 control pigs, the hematoma resorption followed its natural course. The rate of hematoma reduction and edema formation over 10 days was evaluated on planimetry of the MRI data and correlated to the histopathological changes found at autopsy. Although rt-PA significantly accelerated clot resolution compared to controls (p < 0.02), the increase of perihematomatous edema volume within 10 days was not significantly ameliorated in rt-PA-treated animals compared to controls on MRI. The extent of inflammatory infiltrates on histology was more pronounced in animals treated with rt-PA. In conclusion, despite significant reduction in the size of the hematoma clot liquefication with rt-PA and aspiration invokes a substantial inflammatory response when studied after 10 days and does not result in a reduction of the perihematomatous edema. PMID- 12739234 TI - Protective action of recombinant neurturin on dopaminergic neurons in substantia nigra in a rhesus monkey model of Parkinson's disease. AB - Parkinson's disease (PD) is a neurodegenerative disease characterized by muscular trembling palsy due to lack of dopamine (DA) in the substantia nigra-striatum (nigrostriatal) system resulting from the degeneration and necrosis of dopaminergic neurons. No effective cure has been found. Neurturin (NTN) has been demonstrated to act specifically on midbrain (mesencephalic) dopaminergic neurons with protective actions specifically. In the present study, we induced rhesus monkey model of Parkinson's disease by injection of 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP). Rhesus monkeys were randomly divided into a PD model group, NTN treatment group and normal control groups. In the NTN treatment group, 1 mg of E. coli-derived recombinant human NTN was injected into the cerebral ventricles 48 h before the injection of MPTP. Results indicated that Rhesus monkeys in the PD model group acquired PD symptoms that increasingly aggravated over time, while monkeys treated with NTN had less apparent or no symptoms. Using fluorospectrophotometry, the dopamine (DA), 5, 5-hydroxytrytamine (5-HT) and the 5-hydroxyindoleacetic acid (5-HIAA) contents of DA, 5-HT and 5-HIAA in substantia nigra, putamen and caudate nucleus in monkeys from the model group was found to be significantly lower than in the normal control group. While no significant differences were found between monkeys treated with NTN and normal control groups, the contents of DA, 5-HT and 5-HIAA in the NTN treatment group were higher than those observed in the PD model group. A dramatic loss of neurons in the substantia nigra in monkeys in the PD model group was observed by light microscopy, while no obvious loss was observed in the NTN treatment group in which the numbers of neurons were similar to those in normal controls. These results indicate that recombinant human NTN can prevent PD symptoms as well as protect dopaminergic neurons and preserve DA content in midbrain substantia nigra in rhesus monkeys exposed to MPTP. PMID- 12739235 TI - Vasospasm following subarachnoid hemorrhage: evidence against functional upregulation of protein kinase C constrictor pathway. AB - This study tested the hypothesis that vasospasm due to subarachnoid hemorrhage involves the functional upregulation of protein kinase C. Spasm of the rabbit basilar artery was achieved using a double hemorrhage model, which we previously demonstrated was endothelin-1 dependent. In situ effects of agents were determined by direct measurement of vessel diameter following their suffusion in a cranial window. Chelerythrine, a protein kinase C inhibitor, relaxed the spasm. However, relaxations to chelerythrine were not significantly greater in endothelin-1 constricted spastic vessels initially relaxed with the endothelin converting enzyme inhibitor, phosphoramidon, as compared to endothelin-1 constricted control vessels. These results suggest that subarachnoid hemorrhage induced vasospasm does not involve functional upregulation of protein kinase C. PMID- 12739236 TI - Glioma cell-associated sustained activation of the transcription factor, nuclear factor-kappa B, was inhibited by neomycin. AB - Experimental evidence suggests that the transcription factor nuclear factor-Kappa B (NF-kappa B) plays an important role in tumor cell invasion, apoptosis suppression and growth. Malignant glioma is one of the most intractable tumors because of its invasiveness to surrounding brain tissue. Our study investigated the role of neomycin on NF-kappa B activity in glioma cell cultures. We performed immunocytochemical analysis of cells with the antibody NF-kappa Bp65 which results show that neomycin decreases significantly the activation of NF-kappa B when added to glioma cultures for 30 min. This finding supports an important role for neomycin in glioma invasion, apoptosis and growth. Collectively, these data suggest a rationale for clinical trials with neomycin in the treatment of gliomas. PMID- 12739237 TI - Targeting of post-ischemic cerebral endothelium in rat by liposomes bearing polyethylene glycol-coupled transferrin. AB - To achieve an efficient delivery targeting to post-ischemic cerebral vascular endothelium, PEG-liposome conjugated with transferrin (Tf) (Tf-PEG-liposome) was intravenously administered to the rats after 90 min of transient middle cerebral occlusion. The expression of Tf receptor (TfR) in the cerebral endothelium increased with a peak at 1 day after the reperfusion and returned to the control level by 6 days. The Tf-PEG fluorescence was marginally detectable in sham control brain, but remarkably increased with a peak at 2 days, showing about 70% of TfR positive vascular endothelium double-labeled with Tf-PEG. These results indicate that the Tf-PEG-liposome could be utilized as an efficient drug delivery tool to the brain after stroke. PMID- 12739239 TI - MRI and MRS of Coffin-Lowry syndrome: a case report. AB - MRI and MRS were used to examine the brain and the spine of a Coffin-Lowry syndrome (CLS) patient. There were moderately enlarged lateral and third ventricles and subarachnoid space with prominent Virchow-Robin spaces. MRS of basal ganglia and periventricular white matter was normal. PMID- 12739238 TI - An improved functional neurological examination for use in nonhuman primate studies of focal reperfused cerebral ischemia. AB - There is renewed interest in primate models of acute stroke for the evaluation of potential therapeutic agents prior to clinical trials. The development of more precise functional outcome measures would improve the pre-clinical assessment of neuroprotective strategies. We have constructed a grading scale that utilizes an increased number of goal-oriented tasks to assess both behavior and motor function. The new scoring system is designed to enhance precision and accuracy when compared to existing scales. Twenty-seven male baboons were subjected to 1 h of middle cerebral artery territory occlusion followed by reperfusion. Outcome was evaluated using both a standard neurological function scale and a new task oriented scale. Each scoring system was assessed for reproducibility (inter observer reliability) and for association with radiographic infarct volume. The task-oriented grading system was significantly less variable than the standard outcome measure (p < 0.0001). The task-oriented neurological scale demonstrated stronger correlation with radiographic infarct volume (p < 0.0001) than the standard scale (p < 0.01) and more accurately reflected infarct size in animals with small strokes. Compared to the accepted system for grading neurological function, the task-oriented scale demonstrates improved inter-observer variability and a better association with radiographic outcome measures. Incorporating this refined neurological evaluation into a baboon model of stroke may serve to increase the functional predictive value of pre-clinical studies. PMID- 12739240 TI - Steroid effects on vestibular compensation in human. AB - Vestibular neuritis (VN) rapidly damages unilateral vestibular periphery, inducing severe balance disorders. In most cases, such vestibular imbalance is gradually restored to within the normal level after clinical therapies. This successive clinical recovery occurs due to regeneration of vestibular periphery and/or accomplishment of central vestibular compensation. We experienced 36 patients with VN treated at our hospital, including cases in our previous preliminary report. To elucidate effects of steroid therapy both on the recovery of peripheral function and on the adaptation of central vestibular compensation, we examined caloric test and several questionnaires with two randomly divided groups, 18 steroid-treated and 18 nonsteroid-treated patients, over two years after the onset. These examinations revealed that steroid-treated patients had a tendency of better canal improvements (13/18, 72%) than nonsteroid-treated ones (10/18, 55.6%). However, there was no significant difference between these two groups. In cases with persistent canal paresis, steroid-treated patients (n = 5) reduced handicaps in their everyday life due to the dizziness induced by head and/or body movements and the disturbance of their mood, more effectively than those with nonsteroid therapy (n = 8). These findings suggest that steroid therapy with VN could be effective on not only vestibular periphery but central vestibular system, to restore the balance. PMID- 12739241 TI - Combined MR measurements of magnetization transfer, tissue diffusion and proton spectroscopy. A feasibility study with neurological cases. AB - Magnetic resonance imaging (MRI) of diffusion and magnetization transfer was combined with 1H-spectroscopic imaging (CSI) to evaluate the clinical potential of in-vivo profiles of various brain pathologies. Ten patients (multiple sclerosis, cerebrovascular disease, leukodystrophy, Alzheimer dementia) and five healthy volunteers were investigated with diffusion-weighted MRI, magnetization transfer imaging, and CSI. Proton spectra were analyzed as ratios of NAA/Cr and Cho/Cr calculated from the peak areas of N-acetylaspartate (NAA), (phospho) creatine (Cr) and choline (Cho). The apparent diffusion coefficient (ADC) and the magnetization transfer ratio (MTR) were determined in identical voxels to ensure identical partial volume effects compared to CSI. Compared to MTR and ADC assessments, the lower spatial resolution of CSI clearly indicates a hindrance at 1.5 T. In most demyelinating lesions, NAA/Cr reduction paralleled attenuated MTRs and elevated ADCs. By contrast, in acute stroke and some acute MS lesions the ADC was reduced, while MTR and NAA/Cr were also decreased. In Alzheimer's dementia, ADC was increased, MTR unchanged and Cho/Cr increased. In a case of leukodystrophy, ADC was pronouncedly increased, MTR and NAA/Cr both reduced, and Cho/Cr normal. Combined measurements of ADC, MTR and CSI are feasible and provide differential in-vivo information on various brain pathologies. PMID- 12739242 TI - Temocapril prevents motor neuron damage and upregulation of cyclooxygenase-II in glutamate-induced neurotoxicity. AB - To examine the possible neuroprotective effect of temocapril, one kind of angiotensin-converting enzyme inhibitor, against glutamate-induced neurotoxicity, we analyzed the pharmacologic utility of temocapril in a post-natal organotypic culture model of motor neuron degeneration. Treatment with 10(-5) M of glutamate resulted in a motor neuron loss and decreased activity of choline acetyltransferase (ChAT). Cotreatment of 10(-5) M of glutamate and temocapril revealed protective effect on motor neuron death and decreased activity of ChAT. Next we performed reverse transcription-PCR analysis for cyclooxygenase-II (COX II). COD-II mRNA was upregulated in glutamate-treated culture. Cotreatment with temocapril and glutamate inhibited upregulation of COX-II. Taken together, temocapril may have therapeutic potential for diseases which associate with upregulation of COX-II, in addition to its role in glutamate excitotoxicity. PMID- 12739243 TI - Motor evoked potentials in rats with congenital hydrocephalus. AB - Motor evoked potential (MEP) by focal transcranial magnetic stimulation was used to test the functional integrity of the motor cortex in congenital hydrocephalic rats. Magnetic MEPs, using a figure-eight coil above the head, were recorded in the tibialis anterior muscle. The latency of transcranial magnetic MEP was 3.4 msec in nonhydrocephalic rats. In the hydrocephalic rats, the MEP had a lower threshold than in nonhydrocephalic rats, and showed two peaks. Latencies of early and late peaks were 3.9 msec and from 5.4 msec to 10.0 msec, respectively. Our findings suggest that hydrocephalus in rats is associated with changes in pyramidal cell excitability in the motor cortical area, probably induced by the fluctuations in cortical excitability and synaptic interaction in hydrocephalic rats. PMID- 12739244 TI - Assessment of ophthalmic artery collateral pathway in the hemispheric cerebral hemodynamics in patients with severe unilateral carotid stenosis. AB - The role of ophthalmic artery collateral pathway in hemispheric hemodynamics in patients with severe carotid stenosis is controversial. The aim of the present study was to address this question comparing the asymmetry of the velocity in middle and anterior cerebral arteries (MCAs and ACAs) and cerebrovascular reactivity (CVR) in MCA on stenotic side in the patients with unilateral severe stenosis of internal carotid artery (ICA) in patients with and without ophthalmic artery collateral pathway. The cohort of 118 patients with carotid stenosis was prospectively assembled. Fifty patients who had severe unilateral ICA stenosis (71%-99%) by Duplex Ultrasound (DUS) were observed by transcranial Doppler (TCD). Cerebral blood flow velocity in MCA and ACA in both sides, direction of blood flow in ophthalmic artery (OA) and CVR on the side of stenosis were determined. There were 14 patients with retrograde blood flow in OA (Group I). The remaining 36 patients with anterograde flow in OA composed Group II. The degree of interarterial asymmetry of peak and mean velocity (Vpeak and Vmean) in MCA and ACA and CVR in MCA were compared in both groups. The degree of ACA asymmetry by Vpeak was 44.0% +/- 6.9% in Group I and 38.3% +/- 3.9% in Group II (p = 0.49), by Vmean 40.3% +/- 6.7% and 36.6% +/- 3.8% (p = 0.63) respectively. The degree of MCA asymmetry by Vpeak was 24.2% +/- 2.8% in Group I and 19.5% +/- 5.0% in Group II (p = 0.42), by Vmean 23.5% +/- 2.9% and 20.6% +/- 5.1% (p = 0.63) respectively. CVR in Group I was 26.1% +/- 6.1%, in Group II 29.0% +/- 6.7% (p = 0.65). The ophthalmic collateral pathway has no influence on hemispheric cerebral hemodynamics in patients with severe unilateral carotid stenosis. PMID- 12739245 TI - Glutamate enhances caspase-3 immunoreactivity in cultured spinal cord neurons of newborn rats. AB - The role of glutamate in the mechanism of spinal neuron death is not fully understood. With addition of glutamate to primary culture of 11-day-old rat spinal cord, the number of caspase-3 positive small neurons of the dorsal horn greatly increased at 6-24 h in contrast to the case with vehicle. The addition of glutamate made caspase-3 immunoreactivity stronger in the cytoplasm of large motor neurons in the ventral horn. The present results show that excessive amount of glutamate enhances apoptotic pathway through caspase-3 in cultured spinal neurons of newborn rat. PMID- 12739247 TI - The role of the workplace in your transition from student to nurse. PMID- 12739248 TI - From student to nurse. Six tips for a successful transition. PMID- 12739249 TI - The importance of mentoring in the transition from student to nurse. PMID- 12739250 TI - What the Army has taught me about leadership. PMID- 12739252 TI - Mammography screening in the county of Fyn. November 1993-December 1999. AB - This report covers the outcome of the first three invitation rounds of the organised mammography screening programme in the county of Fyn. The programme started in November 1993, and the third invitation round ended on 31 December 1999. The screening takes place either at a special clinic located at University Hospital Odense or in a mobile unit. Women living in and around the city of Odense are examined at the clinic (about 55%), while the rest are examined in the mobile unit. Two-view mammography is used at the first screening. Women with dense breast tissue will continue to have two-view mammography (about 60%), whereas the rest will have singleview mammography at the subsequent screens. All screening images are exposed at the mammography-screening clinic and evaluated with double reading in the clinic. The programme targets women aged 50-69, except those undergoing treatment for breast cancer or going for regular check-ups following breast cancer. Based on the updated population register, the IT-Centre of the county of Fyn issues the invitations. Invited are all women aged 50-69 and living in the county of Fyn when their general practitioners' patients are invited. During the first 3 invitation rounds, 136,079 screening tests were made. Of these, 129,375 tests were made in the women aged 50-69 targeted by the programme. In addition, 6682 screening tests were made in women aged 70 and above, and 22 screening tests were made in women below the age of 50. As a consequence of the mammography screening 2657 assessments were made, 1145 women had surgery, 782 women were diagnosed with invasive breast cancer, and 109 women were diagnosed with ductal carcinoma in situ. A participation rate for the first invitation round was calculated immediately after the end of the round based on the number of participants divided by the number of women invited. This percentage was 88%. Invitation data are, however, not stored. It is therefore not possible now to calculate the participation rates in previous invitation rounds based on the same method. We have therefore chosen to calculate the participation rate as the coverage, i.e. the number of participants divided by the average number of women in the county of Fyn during a given invitation round. Calculated in this way, 84% participated in the first round, 84% in the second round, and 82% in the third round. It should be remembered that these figures do not take into account that some women are not invited because they 1) were undergoing current treatment for breast cancer or going for regular check-ups following breast cancer, or 2) did not participate in the previous round (and never actively informed the programme that they wanted an invitation to the next invitation round), relevant only for the second and third invitation round. For the second and third invitation rounds, the programme only invited women who participated in the previous invitation round, asked the clinic for an invitation, or entered the target population since the last invitation round. Therefore the participation rate in the second invitation round among actually invited women will be close to 94%, as 94% of those participating in the first round came for the second round. For the third invitation round, the participation rate among actually invited women will be close to 96%, as 96% of those participating in the first and second rounds came for the third round. One per cent of the participants in the first invitation round were diagnosed with invasive breast cancer or ductal carcinoma in situ. The detection rate was 0.5% in both the second and third invitation rounds. Ductal carcinoma in situ cases constituted 14% of the detected cases in the first and second rounds, and 10% in the third round. The percentage of invasive breast cancer 10 mm of less was 38%, 31%, and 32%, respectively, and 68%, 74%, and 73%, respectively, were node negative. The screening programme of the county of Fyn fulfilled all the quality assessment parameters specified by the European guidelines on breast cancer screening, except two. The proportionate interval cancer rate was higher than specified in the guidelines, probably mainly due to the fact that the Fyn programme operates without early recalls. The proportion of stage II+ cancers was higher than specified in the guidelines, which seems, however, to be due to inconsistency between some of the performance indicators in the European guidelines. This analysis of the outcome from the first three invitation rounds of the mammography screening programme in the county of Fyn thus showed that it is a programme of high quality with a favourable profile of the prognostic indicators. The screening programme is hopefully well on its way to reducing breast cancer mortality in the county of Fyn. PMID- 12739251 TI - Immune selection in murine tumors. Ph.d thesis. AB - It must be assumed that all tumor cells produce proteins which do not belong to a normal cell. These are called tumor-associated or tumor-specific antigens. In the classic immune surveillance theory it is believed that the cellular immune defense (the T-cell system) continuously discovers and eliminates newly arisen tumor cells which express such tumor-specific antigens. Since then it has been shown that one of the preconditions for the T-cell system to be able to recognize antigens is that they are presented by MHC class I histocompatibility antigens. There is a continual processing and presentation of all intracellular proteins in a cell. Thus, a tumor cell which produces an abnormal protein will also present this and thereby expose itself to being killed by cytotoxic T cells. The antigens are presented in the form of short peptides (8-9 aminoacids), which arise as a result of controlled degradation of the original proteins. The peptides thus formed are transported by specialised molecules in the so-called endogenous antigen processing and presentation pathway, and are eventually bound to and presented by MHC class I molecules. It has been shown that many tumors express less MHC class I on their surface compared to the normal tissue from which they have arisen, and also that patients with reduced immune function have an increased incidence of certain forms of cancer. It is therefore widely believed that a low MHC class I level contributes to the ability of tumor cells to avoid the T-cell-mediated immune defense. The aim of the present research project was to confirm the existence of a T-cell-mediated immune selection in primary tumors. Another of its goals was to elucidate the extent to which tumor cells with low MHC class I expression showed poor ability to present antigen, and whether the reason for this could be found in one or more of the molecular systems which participate in antigen processing and presentation. By using the chemical carcinogen 3-methylcholanthrene a total of 144 tumors were induced in immunologically normal and T-cell defective mice, respectively. It was assumed that tumors induced in normal mice would be immune selected, whilst this would not be the case for tumors from T-cell defective mice. This enabled us to work with a tumor-material where the two populations only differed in that the one part had undergone selection by a T-cell system and the other had not. Tumor induction time turned out to be shorter in immune defective than in normal mice, and the tumor frequency was higher, which might be due to the fact that in normal mice tumor growth was inhibited and in certain cases stopped by the T cells. On transplantation of the uncloned cell lines which were established from the primary tumors to immunologically normal congenic recipients, we were able to show that most of the tumors which originated from mice with a functional T-cell system, and which must therefore be assumed to have undergone selection in the primary tumor host, were not immunogenic and were therefore accepted. On the other hand, most tumors which originated from T-cell-defective mice were rejected as a sign that immunogenic tumor cells, assumed to have expressed tumor antigen, had not been eliminated in the primary tumor host. Still, we found that the ability of tumor cells to induce an immune response on transplantation was not reflected in their MHC class I expression. Both tumor lines from immunodeficient and normal mice had highly varying MHC class I levels, and contrary to expectations the highest levels were seen in tumor lines from immunologically normal mice. At the same time we found that the expression levels for the three different MHC class I molecules were the same in the individual tumor lines, which might indicate that the three genes are syn-regulated. The MHC class I mRNA content in tumors from normal mice was generally concordant with the surface level of MHC protein. Among the tumor lines from immunodeficient mice, on the other hand, we found several where there was no such agreement, which was taken to indicate that tumor cells with deviant MHC class I gene transcription had not been eliminated, in contrast to in the immunocompetent tumor hosts. The ability of tumor cells to present antigen was investigated by infecting cells with virus and thereafter assessing their ability to function as target cells for virus specific T cells in a cytotoxic test. Their ability to do this varied considerably, but showed a correlation with their MHC class I expression. Among the transplanted tumor lines that were not able to present viral antigen, the majority were accepted, while most of the tumor lines which were rejected on transplantation possessed the ability to present virus. Closer analysis of the composition of proteasomes, heat shock protein content and TAP molecule function, which are all involved in the antigen processing system, did not immediately reveal any defects. Treatment with interferon gamma, which is known to upregulate the transcription of MHC class I and a number of other proteins which are involved in antigen presentation, showed that by far the majority of the tumor lines were able to respond normally. This was also true for the tumor lines which had deviant MHC class I gene transcription and the cells which showed poor ability to present viral antigen. We did find, however, three cell lines which did not respond to interferon gamma, and they all had defective interferon gamma signaling, not because they did not express the interferon-receptor on the surface, but possibly on account of their lacking phosphorylation of an intracellular signal molecule, Stat1. PMID- 12739253 TI - Antibody neutralization of human immunodeficiency virus type 1 (HIV-1). PMID- 12739254 TI - Multisource feedback in the assessment of physician competencies. AB - Multisource feedback (MSF), or 360-degree employee evaluation, is a questionnaire based assessment method in which rates are evaluated by peers, patients, and coworkers on key performance behaviors. Although widely used in industrial settings to assess performance, the method is gaining acceptance as a quality improvement method in health systems. This article describes MSF, identifies the key aspects of MSF program design, summarizes some of the salient empirical research in medicine, and discusses possible limitations for MSF as an assessment tool in health care. In industry and in health care, experience suggests that MSF is most likely to succeed and result in changes in performance when attention is paid to structural and psychometric aspects of program design and implementation. A carefully selected steering committee ensures that the behaviors examined are appropriate, the communication package is clear, and the threats posed to individuals are minimized. The instruments that are developed must be tested to ensure that they are reliable, achieve a generalizability coefficient of Ep2 = .70, have face and content validity, and examine variance in performance ratings to understand whether ratings are attributable to how the physician performs and not to factors beyond the physician's control (e.g., gender, age, or setting). Research shows that reliable data can be generated with a reasonable number of respondents, and physicians will use the feedback to contemplate and initiate changes in practice. Performance may be affected by familiarity between rater and ratee and sociodemographic and continuing medical education characteristics; however, little of the variance in performance is explained by factors outside the physician's control. MSF is not a replacement for audit when clinical outcomes need to be assessed. However, when interpersonal, communication, professionalism, or teamwork behaviors need to be assessed and guidance given, it is one of the better tools that may be adopted and implemented to provide feedback and guide performance. PMID- 12739255 TI - Physicians' and patients' attitudes toward manual medicine: implications for continuing medical education. AB - INTRODUCTION: Manual medicine (MM) is a physical modality infrequently used in primary care clinics. This study examines primary care physicians' experience with and attitudes toward the use of MM in the primary care setting, as well as patients' experience with and attitudes toward MM. METHODS: Surveys were distributed to a convenience sample of physicians (54.3% response rate) attending a 1-week primary care continuing medical education (CME) conference in Kentucky. Similar surveys were also mailed to a random sample of primary care patients (35.3% response rate) living in a service region in which most conference attendees practiced. RESULTS: Similar responses were obtained from physicians and patients. A majority (81% and 76%, respectively) of physicians and patients felt that MM was safe, and over half (56% of physicians and 59% of patients) felt that MM should be available in the primary care setting. Although less than half (40%) of the physicians reported any educational exposure to MM and less than one quarter (20%) have administered MM in their practice, most (71%) respondents endorsed desiring more instruction in MM. The majority of those seeking additional educational exposure (56%) were willing to pay for MM training that included CME credit. DISCUSSION: This survey suggests that primary care physicians feel that there is currently insufficient education in MM. The majority of physicians and patients feel that MM is beneficial, safe, and appropriate for use in a primary care setting. Thus, there may be a rising demand for quality instruction in MM from physical medicine doctors and other licensed therapists who currently practice MM. PMID- 12739256 TI - A needs assessment of medical school faculty: caring for the caretakers. AB - INTRODUCTION: We conducted an assessment of need for faculty development and mentoring in a medical school to guide program planning and use of scarce resources. METHODS: A multifaceted approach included semi-structured interviews, nominal group process, and a 36-item questionnaire to reach all faculty in the school, including senior administrators. RESULTS: With a 72% response rate, we validated the questionnaire and, using principal components analysis, identified and prioritized the eight interpretable subdimensions: personal growth, achieving balance, teaching, professional networking, research, administrative skills, career progression, and diversity/ethics. All groups of faculty prioritized learning for sustaining their vitality, balancing their personal and professional lives, finding meaning in their work, relationships, and personal growth. Senior administrators prioritized the following for faculty: time management, an institutional outlook, teamwork, and improved performance in teaching, research, and clinical practice. Junior faculty expressed the need for mentoring, scholarship, research, and career planning. DISCUSSION: Attention to faculty humanistic needs and the disparity between the perceived needs by faculty and senior administrators may help explain the attrition of faculty in academic medicine. PMID- 12739257 TI - Peer coaching as a technique to foster professional development in clinical ambulatory settings. AB - INTRODUCTION: Few studies have examined how peer coaching is an effective educational and development technique in contexts outside the classroom. This research focused on peer coaching as a platform to study the process of professional development for physicians. The purpose was to identify perceived benefits coaches received from a coaching encounter and how this relates to their own process of professional development. METHODS: Critical incident interviews with 13 physician coaches were conducted and tape recorded. Themes were identified using a thematic analysis technique. RESULTS: Themes emerged clustering around two distinct benefit orientations. Group 1, reflection and teaching coaches, tended to focus on others and discuss how positively they experienced the encounter. Group 2, personal learning and change coaches, expressed benefits along more personal lines. DISCUSSION: Peer coaching contributes to physicians' professional development by encouraging reflection time and learning. Peer coaching affords positive impact to those who coach in addition to those who receive the coaching. The two clusters of benefits support the performance, learning, and development theory in that there are multiple modes to describe adult growth and development. Programs of this type should be considered in medical faculty development activities associated with medical education. PMID- 12739259 TI - Performance in reading radiographs: does level of education predict skill? AB - INTRODUCTION: Previous studies demonstrated lack of progress in electrocardiographic analysis skills with increasing levels of medical education. This study examined radiograph analysis skills, a similar cognitive task, across a range of educational experience: senior medical students (n = 23), family practice residents (n = 16), general practitioners (n = 41), and their family practice educators (n = 7). METHODS: Written records of diagnosis or description of abnormalities were used to detect skill in interpreting radiographs. The instrument was 12 sets of radiographs: 2 normal and 6 abnormal chest radiographs and 2 normal and 2 greenstick radial fractures. RESULTS: The mean score for correctly diagnosing all 12 sets of radiographs was 5.59 +/- 1.68, and the mean score for correctly diagnosing the 4 normal radiographs as normal and the abnormal radiographs as abnormal was 8.76 +/- 1.55. There was no statistically significant difference between the four groups of participants. DISCUSSION: Skill level in interpreting radiographs did not appear to improve with additional experiential training and may require a more formal educational approach to address this issue. PMID- 12739260 TI - Spouse/partner violence education as a predictor of screening practices among physicians. AB - INTRODUCTION: Spouse/partner violence is a major public health problem that affects 3 to 6 million women per year. Many studies show that the majority of health care practitioners do not detect or respond to cases of spouse/partner violence in their practice. Research suggests that there are potential barriers to reporting or detecting this problem. A barrier often cited is lack of proper education or training regarding spouse/partner violence. The objective of this study was to determine if physicians who received spouse/partner violence education at various stages of their careers were more likely to screen patients for spouse/partner violence. METHODS: A survey was developed and administered to family physicians and obstetricians/gynecologists in Virginia. The data were analyzed to determine screening practice and spouse/partner violence education among respondents. Four different educational opportunities were analyzed to determine potential determinants of screening. RESULTS: All respondents who had spouse/partner violence education were more likely to screen every patient than those who were lacking this education. Receiving lectures during residency training was found to be a significant predictor of screening every patient for spouse/partner violence among respondents. DISCUSSION: Screening every patient for exposure to spouse/partner violence is the ideal situation. This study indicates that education about spouse/partner violence has a significant impact on screening tendencies if provided during a physician's residency program. PMID- 12739258 TI - Videoconferencing for practice-based small-group continuing medical education: feasibility, acceptability, effectiveness, and cost. AB - INTRODUCTION: Small-group, practice-based learning is an effective and well accepted method of continuing medical education (CME). However, one limitation is that many physicians work in communities with fewer than the minimum number recommended for an effective learning group. Videoconferencing has the potential to remove this limitation. The purpose of this study was to evaluate the feasibility, acceptability, effectiveness, and cost of conducting practice-based, small-group CME learning by videoconference. METHODS: Through a videoconferencing link, 10 learners in three communities were guided through four practice-based learning modules by a trained facilitator at a fourth site. Data were collected through evaluation questionnaires, direct observation by the research team, pre- and post-knowledge tests, a focus group, and an interview. RESULTS: A total of 31 learners participated in the four modules. Videoconferencing was generally well accepted by learners. The facilitator and research team observers noted that muting microphones, video quality, audio quality, and audio lag all somewhat hindered discussion. Overall, the facilitator found moderating by videoconference only slightly more difficult than a face-to-face session. There was evidence of knowledge gain, with post-test scores being 20% higher than pretest scores (p = .006). Learners reported nine practice changes from taking the modules. At commercial rates, telecommunications costs per videoconferenced module were approximately CAN$1,200. DISCUSSION: Videoconferencing has the potential to bring the benefits of small-group, practice-based learning to many physicians; however, strict attention to videoconferencing techniques is required. Cost is also an important consideration. PMID- 12739261 TI - [Clinical methods in evolutional physiology]. PMID- 12739262 TI - [Paradoxal sides of human thinking]. PMID- 12739263 TI - [Functional asymmetry of the brain and neglect of the environment]. PMID- 12739264 TI - [L.Ya Balonov's and V.L. Deglin's input in studying of speech and thinking activity from the position of functional brain asymmetry]. PMID- 12739265 TI - [About disturbance of the interaction between brain hemispheres in psychopathological conditions (the development of L. Ya. Balonov's and V. L. Deglin's ideas)]. PMID- 12739266 TI - [The development of vision and visual perception in infants]. PMID- 12739268 TI - [The changes in visuo-spatial perception in emotional disorders]. PMID- 12739267 TI - [The study of metaphoric and associative thinking in children of different age groups and in patients with childhood autism]. PMID- 12739269 TI - [Functional asymmetry in estimation of movement trajectory of whole sound by man]. PMID- 12739270 TI - [The role of movement asymmetry and sex in lateralization of autonomous effects]. PMID- 12739271 TI - [Age-related changes in semantic content of associative reactions and the influence on it of the emotional sign of verbal stimulus]. PMID- 12739272 TI - [Speech peculiarities in right brain hemisphere dysfunction]. PMID- 12739273 TI - [The role of neuropeptides in brain asymmetry]. PMID- 12739274 TI - [The changes in the interaction of the brain hemispheres in the course of drug therapy of psychosis]. PMID- 12739275 TI - The war against an unknown pathogen: rising to the SARS challenge. PMID- 12739276 TI - Rotavirus vaccines, an update. PMID- 12739277 TI - When patients present a challenge. PMID- 12739279 TI - What you need to know about ... panic attacks. PMID- 12739278 TI - Assessing CHD and hypertension in minority ethnic communities. AB - Hypertension, coronary heart disease and stroke rates are higher in minority ethnic communities than their European counterparts. A health-needs assessment was undertaken to determine the prevalence of undiagnosed/asymptomatic hypertension among these communities in a primary care setting. Preliminary results indicate the prevalence to be 32.49 per cent. A number of strategies are discussed for improving the health care of minority ethnic communities and reducing health inequalities. PMID- 12739280 TI - Health assessment. PMID- 12739281 TI - Non-steroidal anti-inflammatory drugs: indications for use. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) are among the most widely used of all therapeutic agents. Although they are effective analgesics they do have significant adverse side-effects. Before treatment is started the prescriber should always weigh efficacy against possible side-effects. NSAIDs vary in their selectivity for inhibiting the two types of cyclo-oxygenase. An NSAID should therefore be chosen on the basis of the incidence of associated side-effects. PMID- 12739282 TI - Education to achieve symptom control for patients with cancer. AB - This project evaluated whether shared interprofessional education across primary and secondary care could improve symptom control for patients with cancer. An action learning group of 16 health care professionals and patients from the community and hospital took part in an initial three-month learning and change process and an evaluation at the end of one year. Both participants and facilitators developed skills in working with other health care settings, as well as experience of using a continuous quality improvement tool. Participants developed a greater understanding of how the NHS worked and what was required to enable patients to receive improved care. The project enabled professionals to work more effectively with both primary and secondary services, agencies, and helped patients to enhance symptom control. PMID- 12739283 TI - Building hope, breaking taboos. Interview by David Crouch. PMID- 12739284 TI - When the shooting stops. PMID- 12739285 TI - One-to-one care in a children's hospice. PMID- 12739286 TI - Shining examples. PMID- 12739287 TI - Slipping through the net. PMID- 12739288 TI - What is nursing? PMID- 12739289 TI - Lay it on the line. PMID- 12739290 TI - Practical issues of student assessment. AB - Student nurses are required to undergo assessment in their practice placements as part of their pre-registration programme. This process is ongoing throughout the programme and will require students to be flexible in their many interactions with these areas. This involves the need to provide evidence of their learning, together with an ability to apply this in practice. This article identifies the need for all those involved in this process to take account of the human variables that inevitably arise and incorporate strategies to ensure that the outcome is rigorous, while remaining a positive experience. PMID- 12739291 TI - Tuberculosis: protecting healthcare workers. AB - The global incidence of tuberculosis (TB) is rising. Many more healthcare workers are coming into contact with the disease now that it has reached its highest infection rate for 17 years (Sadler 2001). Modern screening methods allow detection of infection, and a strict antibiotic regimen can prevent and cure disease. A clinical placement at a chest clinic in New South Wales (NSW), Australia, provided a useful insight into methods used for the screening and control of TB, and allowed a comparison and analysis of how infection is monitored and controlled among healthcare workers. Although approaches and procedures differ, this information provides scope for analysis and discussion, which can be shared to achieve the best outcome locally, nationally and internationally to protect citizens against this increasing global communicable disease. PMID- 12739292 TI - Travel advice for clients with pre-existing medical conditions. AB - Overseas travel, especially long-haul, can have risks that are not only associated with infection but also the mode of transport, climate, lifestyle and conditions at the destination. These risks may be magnified if the individual has a pre-existing medical condition. PMID- 12739293 TI - Occupational irritant contact dermatitis. AB - This article raises awareness of the risks of irritant contact dermatitis among different occupational groups, especially healthcare professionals, and suggests strategies to aid prevention. PMID- 12739294 TI - From task to talking in wound care. AB - The author outlines a model that can be used to separate nursing tasks from talking to patients. It is hoped that the model will promote effective use of time and improve the quality of nurse-patient interactions in wound care and other clinical settings. PMID- 12739295 TI - The complexities of heel ulcers. AB - This article examines the complexities of heel ulcers. Treatments now considered outdated and potentially hazardous are discussed as are modern treatments that may reduce the development of heel ulcers. PMID- 12739296 TI - Health issues of migrant and seasonal farmworkers. AB - This paper describes the socioeconomic conditions under which the 3 to 5 million migrant and seasonal farmworkers in the United States live. Health consequences resulting from occupational hazards and from poverty, substandard living conditions, migrancy, language and cultural barriers, and impaired access to health care are described. Specific problems include infectious diseases, chemical- and pesticide-related illnesses, dermatitis, heat stress, respiratory conditions, musculoskeletal disorders and traumatic injuries, reproductive health problems, dental diseases, cancer, poor child health, inadequate preventive care, and social and mental health problems. By increasing awareness among health care professionals of the plight of migrant and seasonal farmworkers, the authors hope to encourage development of a stronger public health infrastructure and to improve the health status of these individuals. PMID- 12739297 TI - Free clinics and the uninsured: the increasing demands of chronic illness. PMID- 12739298 TI - The prevalence of physicians who emphasize wellness and provide educational materials for their patients. PMID- 12739299 TI - Organ donation among Asians and Pacific Islanders: preliminary research findings. PMID- 12739300 TI - Community care for people who are homeless and mentally ill. AB - This qualitative longitudinal study documents the experiences of 60 people who are homeless and mentally ill from their state mental hospital discharge through their first two years in community housing. The study explores the personal, cultural, and environmental contexts of life for adults who are homeless and mentally ill and examines the interaction between an individual's needs and community resources. The research identifies forces that perpetuate homelessness and traces the struggles that people who are homeless and mentally ill encounter during the transition from the streets to stable housing. The findings describe a culturally based pattern of mutual avoidance between homeless mentally ill clients and caregivers, which limits delivery of services to the population. Recommendations include development of alternative systems of care delivery, expansion of educational experiences with underserved populations, and increased funding for service or research with people who are homeless and mentally ill. PMID- 12739301 TI - Adult health care access and use under Medicaid: does it vary by state? AB - States are given considerable discretion in designing their Medicaid programs and, as a result, Medicaid is made up of 51 different programs. Using data from the 1997 National Survey of America's Families, the authors examine variation in health care access and use by adults in 13 states and then compare those differences with national estimates of access and use. The authors find significant differences in access and use among the 13 study states. This variation persisted after differences in state Medicaid caseload characteristics, including demographic, socioeconomic, and health status, were controlled for. State variation in beneficiaries' access and use also persisted after differences in health care market characteristics, such as supply of health care providers, level of employer-sponsored coverage, and level of HMO penetration, and characteristics of the state Medicaid programs were controlled for. PMID- 12739302 TI - SSI enrollees' health care in TennCare. AB - How well does TennCare, Tennessee's Medicaid managed care program, meet the needs of blind/disabled Supplemental Security Income (SSI) enrollees? People with disabilities have extensive health care needs and greater barriers to accessing care, so efforts to reduce service use may decrease their health and independence. On the other hand, managed care plans may better coordinate care. Computer-assisted telephone surveys of urban SSI and other urban TennCare enrollees were conducted to assess these issues. SSI enrollees in TennCare had mixed experiences, and they faced problems in areas particularly important to people with disabilities. Relative to other TennCare enrollees, SSI enrollees had similar or slightly worse access to care and satisfaction. A significant minority of SSI enrollees reported unmet needs for care, such as not getting referrals to specialists, prescription drugs, and special medical equipment. Lack of care coordination was a problem for some SSI enrollees. PMID- 12739303 TI - Discontinuation from HIV medical care: squandering treatment opportunities. AB - The purpose of this study was to assess HIV-infected patients' discontinuation of their primary care. One hundred ninety-eight consecutive outpatients were interviewed on initial HIV primary care presentation, assessed after six months about their discontinuation from primary care, and had characteristics associated with discontinuation determined. Primary care was not continued in 20 percent (40/198) of the cases. Cohort characteristics included 25 percent women; 44 percent black, 28 percent white, 25 percent Hispanic; 69 percent with highest yearly income < or = $16,000; 47 percent injection drug users; and median CD4 count 285/microL. Characteristics significantly associated (p < or = 0.05) with discontinuation were higher CD4 count, less education, no history of victimization, previous jail time, and site of medical care. One-fifth of HIV infected patients did not remain engaged in primary care after establishing this essential link to treatment. PMID- 12739305 TI - Prenatal care and infant birth outcomes among Medicaid recipients. AB - Infant morbidity due to low birth weight and preterm births results in emotional suffering and significant direct and indirect costs. African American infants continue to have worse birth outcomes than white infants. This study examines relationships between newborn hospital costs, maternal risk factors, and prenatal care in Medicaid recipients in an impoverished rural county in South Carolina. Medicaid African American mothers gave birth to fewer preterm infants than did non-Medicaid African American mothers. No differences in the rates of preterm infants were noted between white and African American mothers in the Medicaid group. Access to Medicaid services may have contributed to this reduction in disparities due to race. Early initiation of prenatal care compared with later initiation did not improve birth outcomes. Infants born to mothers who initiated prenatal care early had increased morbidity with increased utilization of hospital services, suggesting that high-risk mothers are entering prenatal care earlier. PMID- 12739306 TI - Definition, epidemiology, course, and prognosis of COPD. AB - Chronic obstructive pulmonary disease (COPD) is now recognized as our nation's most rapidly growing health problem. It ranks as the 4th most common killer and is the only disease in the top 10 whose rank is rising. In 2000, more women than men (59,936 vs 59,118) died of COPD (1). The National Heart, Lung, and Blood Institute has calculated that in 2001, COPD was a $34.4 billion burden on society (both direct and indirect costs) (2). Two new initiatives, the National Lung Health Education Program (NLHEP) (3,4) and the Global Initiative for Chronic Obstructive Lung Disease (5), promote the early diagnosis and intervention of COPD. Both initiatives offer guidelines for the care of patients with all stages of COPD. The NLHEP recommends spirometry in all current or former smokers age > or = 45 years and anyone with symptoms of chronic cough, excessive dyspnea on exertion, or wheezing (6). "Test your lungs, know your numbers" is the motto of the NLHEP. Most patients with COPD are first seen by their primary care practitioner well before symptoms or signs of moderate-to-advanced stages of the disease are present. Thus, the primary care practitioner, working on the front line, is in the position to make a difference in the treatment and outcome of this devastating disorder. PMID- 12739304 TI - Caring for the underserved: current practice of alumni of the National Health Service Corps. AB - The objective of this study was to examine the number and characteristics of National Health Service Corps (NHSC) alumni who currently practice with an underserved population. The study design was a cross-sectional survey mailed in 1998 to a sample of 2,160 alumni. The response rate was 58.9 percent; overall, 52.5 percent of respondents reported currently working with the underserved. In bivariate analyses, retention in practice with the underserved was associated with several clinician and service experience variables. In a logistic regression model, higher initial desire to work with the underserved, older age, job satisfaction, and higher salary were associated with current service. The authors conclude that more than half of the NHSC alumni responding to the survey reported working with an underserved population in 1998. Associations between clinician and service experience characteristics and retention in practice with the underserved can inform policy and research to improve retention. PMID- 12739307 TI - Pathogenesis of COPD. AB - Chronic obstructive pulmonary disease (COPD) is not a single disorder. Rather, it is a collection of disorders characterized by the progressive development of airflow limitation. Patients afflicted with COPD experience systemic effects that are often responsible for major clinical problems. Lesions with differing pathogenetic bases can contribute to airflow limitation, and these differing and overlapping pathogenetic mechanisms account for the heterogeneous clinical features of COPD. PMID- 12739308 TI - Management of the symptomatic patient. AB - Chronic obstructive pulmonary disease (COPD) is both preventable and treatable. There has been a significant reduction in premature morbidity and mortality from heart disease and stroke, the number 1 and 3 killers in the United States, respectively, largely because of efforts to increase both clinician and public awareness of these diseases. Early and aggressive management of COPD has not yet reached the same level. Primary care professionals are in the forefront in the early detection, prevention, and treatment of this serious national health problem. PMID- 12739309 TI - Treatment of acute exacerbations of COPD. AB - Chronic obstructive pulmonary disease (COPD), the fourth leading cause of death in the United States, is increasing worldwide and is projected to be the third leading cause of death in the United States by the year 2020 (1). It affects nearly 16 million Americans, and more than $18 billion is spent annually on medications, physician visits, and hospitalizations. COPD is characterized by chronic airflow obstruction with episodic acute exacerbations, which result in increased morbidity and mortality. Patients hospitalized with exacerbations have an overall mortality rate of 3% to 4%, and up to 24% of patients requiring care in the intensive care unit die (2). Since forced expiratory volume in 1 second correlates closely with life expectancy and exacerbation rate, early diagnosis (through spirometric testing) and prevention may reduce acute exacerbations and health care costs. PMID- 12739310 TI - COPD in VA hospitals. AB - The Department of Veterans Affairs (VA) Health Care System is the largest integrated single-payer system in the United States. Its primary mission is to provide primary care, specialized care, and related medical and social support services to veterans. Much time and resources are expended on chronic obstructive pulmonary disease, known as COPD, at VA hospitals and clinics, thereby justifying the development of multifaceted strategies to address this problem. This article discusses the special problems of COPD in veterans who use VA facilities. The article also highlights the contributions of the VA to the research, training, and development of clinical practice guidelines for the management of this pervasive disease and presents the challenges that threaten its role in this area. PMID- 12739311 TI - Future treatment strategies for COPD. AB - As the fourth leading cause of death in the United States, chronic obstructive pulmonary disease (COPD) is receiving increased attention from medical researchers. The Global Initiative for Chronic Obstructive Lung Disease now defines COPD as a progressive disease state characterized by chronic respiratory tract inflammation as well as airflow limitation that is partially reversible with appropriate treatment. The inflammatory pathway associated with COPD, although not well studied in the past, is also receiving more attention as medical researchers attempt to elucidate the cellular and molecular pathogenetic mechanisms that result in the development of COPD. Understanding the pathogenetic mechanisms will lead to new therapeutic strategies. The purpose of this review is to identify the known major therapeutic targets and the agents available to treat this devastating disease and some of the new agents that are being developed. PMID- 12739312 TI - Pathophysiology, epidemiology, and impact of migraine. AB - Despite a decade of progress, migraine headache remains prevalent, disabling, underdiagnosed, and undertreated in the United States. Migraine affects approximately 12% of the population, and the economic burden in terms of annual cost of labor lost to migraine disability is between $5.6 and $17.2 billion. The threshold for migraine may be genetically determined, although recent genetic and neurophysiologic studies point to migraine as possibly a channelopathy. Cerebral cortical and brain stem changes occur in migraine. Head pain and associated symptoms of migraine can be explained by activation of the trigeminal vascular system. Evidence has also been accumulated that suggests the release of nitric oxide is an important trigger mechanism. Introduction of the triptans has dramatically advanced acute migraine pharmacotherapy, and preventive therapy has greatly improved; however, public health initiatives may be needed to further advance diagnosis and treatment of this common and disabling disorder. PMID- 12739313 TI - Clinical manifestations of migraine. AB - Migraine is a disorder with many clinical manifestations that vary from person to person and often from headache to headache in the same person. Several body organ systems are involved in a migraine attack. It is a condition that is underdiagnosed by physicians and misidentified by persons suffering with headaches. Many individuals with migraine self-diagnose as having sinus headaches or tension headaches. This article discusses the many symptoms of migraine and will help the busy practitioner to better recognize and diagnose this common disorder. PMID- 12739314 TI - Pitfalls in migraine diagnosis and management. AB - The decade of the 90s brought about a revolution in our understanding of migraine and in medications to treat this disorder. Indeed, our understanding of the pathophysiology of migraine and the pharmacology involved in its treatment has exploded. Nonetheless, there are still many pitfalls that may occur in the clinical assessment and management of migraine that may prevent the patient from receiving the full benefits of the advances of the past 10 years. This article explores the reasons for the unsuccessful diagnosis and treatment of migraine and presents suggestions for improving its management. PMID- 12739315 TI - Acute and prophylactic management of migraine. AB - Although considerable advances have been made in the understanding and treatment of migraine, uncertainty about the management of this disorder remains. Once a diagnosis is established, an effective treatment plan should be developed that includes education for the patient about migraine and reassurance that the headaches are not caused by structural pathology, stress, or psychopathology. Rapidly effective acute treatment, nonpharmacologic treatment such as behavioral and lifestyle changes, preventive measures, and patient follow-up are other essential elements of migraine management. PMID- 12739316 TI - Comparative aspects of triptans in treating migraine. AB - Triptans, beginning with sumatriptan, have revolutionized the treatment of migraine. New triptans in several formulations will soon become available in the United States. Although the similarities of these 5-hydroxytryptamine (5-HT) 1B/1D receptor agonists outweigh their differences, important differences in pharmacokinetics and clinical responses do exist. Subcutaneous sumatriptan has the most rapid onset of action and greatest efficacy but the most adverse effects. Intranasal sumatriptan also has rapid onset of action, but at 2 hours its efficacy is comparable to that of oral zolmitriptan. Of the oral triptans, rizatriptan seems to have the greatest early efficacy. Both rizatriptan and zolmitriptan are now available as rapidly dissolving wafers. Almotriptan, the newest of the triptans, has a response rate similar to that of oral sumatriptan and may produce fewer adverse effects. Naratriptan and frovatriptan, with their slow onset, high tolerability, and long half-lives, may have a role in aborting prolonged migraine attacks and in headache prevention. Eletriptan at higher doses (80 mg) has a response rate approaching that of rizatriptan but may be limited by potential side effects. The many triptans available offer the opportunity to individualize migraine treatment, depending on the patient's attack characteristics, tolerance, and preferences. PMID- 12739317 TI - An approach to clinical dermatologic diagnosis based on morphologic reaction patterns. AB - How do dermatologists examine a patient and instantly generate a differential diagnosis? Of course, as in any other field, experience plays a large role. After a condition with a distinctive appearance is seen hundreds of times, recognition becomes automatic. Nevertheless, dermatologists need to employ a disciplined mental process to generate a differential diagnosis from clinical findings. From the perspective of clinical diagnosis, all dermatology falls into 2 separate categories--inflammatory and neoplastic. The reaction pattern technique discussed in this article relates only to inflammatory conditions such as psoriasis, eczema, and urticaria. This article describes an approach to differential dermatologic diagnosis based on clinical reaction patterns. PMID- 12739318 TI - Acne vulgaris and rosacea: evaluation and management. AB - Acne vulgaris, commonly termed acne, is an extremely common disease. It can be found in nearly all teenagers to some degree as well as in women in their 30s. Regardless of severity, acne often has a greater psychologic effect than cutaneous effect. Indeed, most patients overestimate the severity of their disease, while most physicians underestimate its impact on their patients. Studies have shown that people with severe acne as teens are less employable as adults and that self-esteem is low. When combined with other adolescent tensions, acne can be a difficult disease to treat. Rosacea, which usually starts in the late 20s, may affect the eyes as well as the skin. This article describes the pathogenesis of acne and rosacea and treatment approaches the primary care physician can use. PMID- 12739319 TI - Skin cancer: recognition and management. AB - Skin cancer is the most common malignancy in humans and accounts for one third of newly diagnosed cancers--it will be diagnosed in approximately 1 in 5 Americans in their lifetime and > 1 million cases are diagnosed each year. Skin cancer can cause local tissue destruction, disfigurement, and even death if left untreated; therefore, timely recognition, treatment, and appropriate referral are critical to reducing morbidity. As the incidence of skin cancer rises each year, the primary care physician needs to be familiar with the clinical presentation, treatment options, and means of prevention of the most common skin cancers: basal cell carcinoma, squamous cell carcinoma, and malignant melanoma. PMID- 12739320 TI - Topical medications: a focus on antifungals and topical steroids. AB - Because skin disease is accessible, it can be treated with locally applied medication, which offers great advantages--exposure to a drug is limited to the affected skin and systemic effects of potentially toxic drugs are minimized. Ointments, creams, antifungals, and antibiotics all have their place in treating various skin diseases. Topical steroids, the largest group of topical medications, are effective but present the potential for side effects. This article discusses current and new topical medications that can be used to treat a range of skin diseases. PMID- 12739321 TI - Common skin disorders in the elderly. AB - Skin diseases commonly seen in the elderly are more often than not the effects of sun damage or vascular disease. The effects of a lifetime of even casual sun exposure can be dramatic. Chronically sun-exposed skin becomes thin, loses collagen, and has disrupted elastin and decreased glycosaminoglycans. The result is skin that breaks easily, bruises, sags, irritates easily, and itches. The spots and bumps that patients associate with age are all sun-induced. Consider how lesionless a 60-year-old's buttock is compared to the extensor forearm. The reason that bruising attributed to anticoagulation seems to occur exclusively on the extensor forearm and not the volar aspect of the arm is that sun-induced elastin degradation is greatest on the extensor forearm. Even trivial trauma will cause unsupported capillaries to shear and bleed whether the patient is anticoagulated or not. This article reviews the primary skin disorders associated with the elderly and some of the management approaches that the primary care physician can use. PMID- 12739322 TI - Screening for colorectal cancer. AB - Colorectal cancer screening can prevent the development of colorectal cancer and reduce the risk for death. Screening recommendations include fecal occult blood testing, sigmoidoscopy, radiographic imaging of the colon, and colonoscopy. This article focuses on recommendations for average-risk individuals and discusses the potential benefits and limitations of each. High-risk individuals should be screened with colonoscopy. Cost-effectiveness analyses of colorectal cancer screening are summarized. PMID- 12739323 TI - A clinical approach to constipation. AB - The economic impact of constipation is large. The condition prompts an estimated 2.5 million physician visits per year, with 100,000 referrals to gastroenterologists. Almost all (85%) of these physician visits result in a prescription for a laxative. Each year, American spend approximately $800 million on laxatives. For patients referred for diagnostic evaluation, the average cost is approximately $3000, mostly due to the cost of colonoscopy. This article discusses the pathophysiology of constipation and presents a practical approach to evaluating and treating this disorder. PMID- 12739324 TI - Irritable bowel syndrome. AB - Irritable bowel syndrome (IBS) is one of the most common diagnoses made in the primary care setting and is responsible for up to 40% of referrals to gastroenterologists. Approximately 70% of persons who meet the diagnostic criteria for IBS do not seek health care, and the remaining account for 12% of visits to primary care providers. IBS is a functional bowel disordered comprising abdominal pain associated with defecation or a change in bowel habit with features of disordered defecation and distension in the absence of any demonstrable abnormality. The diagnosis is based on clinical findings and the exclusion of other disorders. New pharmaceutical agents are available to treat the underlying disorder; however, the treatment of IBS still involves a comprehensive, multicomponent approach that includes medical management of dominant symptoms, dietary modifications, and, possibly, psychotherapy. PMID- 12739325 TI - Diseases of the rectum and anus: a clinical approach to common disorders. AB - Diseases of the rectum and anus are common, and the prevalence in the general population is probably much higher than that seen in clinical practice since most patients with symptoms referable to the anorectum do not seek medical attention. The examination and diagnosis of certain anorectal disorders can be challenging, and the physical examination of the anorectum is often inadequately performed in clinical practice. This article reviews the important features of the anorectal examination and the diagnosis and treatment of benign anorectal disorders such as hemorrhoids, fissures, fistulas, solitary rectal ulcer syndrome, fecal incontinence, and pruritus ani. Approaches to staging and managing malignant neoplasms of the anus and rectum are outlined. PMID- 12739326 TI - Inflammatory bowel disease. AB - Inflammatory bowel disease (IBD) comprises a group of idiopathic diseases of the intestine characterized by chronic inflammation of the bowel with periods of exacerbation and remission. The 2 major categories of IBD are ulcerative colitis and Crohn's disease, and each disease is distinct both clinically and histologically. The exact cause of IBD remains unknown, but several theories have been proposed. Primary care physicians are frequently confronted with the initial diagnosis and management of patients with IBD, and medical treatment options have rapidly expanded in recent years. This article reviews the epidemiology and pathogenesis of IBD; its clinical presentations and complications, which are essential to understanding this complex disease; and the available treatment options. PMID- 12739327 TI - Management of asthma exacerbations. AB - The 1997 Expert Panel Report 2 from the National Asthma Education and Prevention Program* details principles and goals for managing asthma exacerbations, based on scientific literature and the opinion of the panel. The panel's recommendations are summarized here, along with approaches to the evaluation and management of patients with asthma exacerbations. Methods to assess and classify the severity of asthma exacerbations are discussed, and treatment objectives for mild, moderate, and severe exacerbations are presented, along with a discussion of postinfectious acute airway hyperresponsiveness. A review of pharmacologic agents used in the treatment of asthma exacerbations is also included. Key points in the management of asthma exacerbations include the notion that early treatment is the best strategy for management. Important elements of early treatment include recognition of early signs of worsening asthma, a written action plan to guide patient self-management, appropriate intensification of therapy, and prompt communication between patient and provider about deterioration in asthma control. Other key points include the use of inhaled beta 2-adrenergic agonists to provide prompt relief of airflow obstruction, the early use of systemic corticosteroids for patients with moderate to severe exacerbations or for patients who fail to respond promptly and completely to an inhaled beta 2-adrenergic agonist, and monitoring response to therapy with serial measurements of lung function. PMID- 12739328 TI - Inherited risk factors for venous thromboembolism: implications for clinical practice. AB - Each year, thrombosis in the deep veins of the lower extremities occurs in 1 of every 1000 Americans (1). Although most cases of venous thrombosis are not fatal, death from pulmonary embolism can be expected in 1% to 2% of all patients, and as many as 25% of patients with deep venous thrombosis (DVT) will suffer the chronic effects of postthrombotic syndrome. Thus, identification of high-risk patients with genetic predispositions to thrombosis is an important clinical goal, particularly among individuals with recurrent DVT. PMID- 12739329 TI - Prevention and treatment of osteoporosis. AB - Osteoporosis develops in older adults when the normal processes of bone formation and resorption become uncoupled or unbalanced, resulting in bone loss. Fractures are the result of decreased bone mass and strength and, in the case of wrist and hip fractures, usually involve a fall. Osteoporosis prevention and treatment programs should then focus on strategies that minimize bone resorption and maximize bone formation as well as on strategies that reduce falls. Optimal treatment and prevention of osteoporosis require modification of risk factors, particularly smoking cessation, adequate physical activity, and attention to diet, in addition to pharmacologic intervention. A number of pharmacologic options are now available to health care providers. This article focuses on US Food and Drug Administration-approved medications for osteoporosis and emphasizes the importance of using these agents as part of a comprehensive program that includes nonpharmacologic measures, complete diagnostic evaluation, and adequate follow-up with bone mineral density measurement. PMID- 12739330 TI - Management of advanced heart failure. AB - Congestive heart failure (CHF) due to progressive systolic dysfunction has become a modern-day epidemic. Despite the increased incidence and prevalence, significant progress has been made in the past 10 to 15 years in the treatment of CHF at all stages. The current outlook for patients with newly diagnosed, mild heart failure is encouraging. It should be noted, however, that most of the morbidity and health care expenditure is incurred by a minority of patients diagnosed with CHF who are in the advanced stages of their disease. The thrust of this article will be to provide practical advice beyond current guidelines on the management of advanced CHF. PMID- 12739331 TI - Differential diagnosis of dementia: clinical examination and laboratory assessment. AB - Recent breakthroughs in putative disease-modifying interventions for Alzheimer's disease (AD) underscore the urgency of making the earliest possible diagnosis. In the absence of a convenient and reliable laboratory test for AD, the clinical assessment is still the cornerstone of the diagnostic approach. This article provides a basis for conducting an assessment within the realities of a busy clinical practice for patients complaining of cognitive decline. The assessment will enable the clinician to diagnose the earliest manifestation of AD. PMID- 12739332 TI - [Standards, Options and Recommendations 2002 for the use of Positron Emission Tomography with [18F]-FDG (PET-FDG in cancerology (integral connection]. AB - CONTEXT: The "Standards, Options and Recommendations" (SOR) project, started in 1993, is a collaboration between the Federation of French cancer centers (FNCLCC), the 20 French cancer centers, scientific societies, and specialists from French public universities, general hospitals and private clinics. The main objective is the development of clinical practice guidelines to improve the quality of health care and the outcome of cancer patients. The methodology is based on a literature review and critical appraisal by a multidisciplinary group of experts, with feedback from specialists in cancer care delivery. OBJECTIVES: To develop clinical practice guidelines for the use of positron emission tomography with [18F]-FDG in oncology according to the definitions of the Standards, Options and Recommendations project. METHODS: More than 600 articles were identified by searching Medline, web sites, and using the personal reference lists of members of the expert groups. Once the guidelines were defined, the document was submitted for review to independent reviewers (n = 82). RESULTS: The different steps in patient care are studied in this report: diagnosis of the primary disease, initial and secondary assessment of metastatic extension, evaluation of treatment response and examination for recurrence. The guidelines were developed for each cancer sites: bronchopulmonary and pleural cancers, melanomas, gynaecological cancers, gastrointestinal cancers, head and neck cancers, urological cancers, lymphomas, soft-tissue and bone sarcomas, cancers of the thyroid and carcinomas of unknown primary site. The recommendations were made on the basis of data published up to February 2002. Systematic monitoring of the new scientific data on FDG-PET has been set up by the working group of the FNCLCC, which will ensure dissemination of the updating on its web site: www.fnclcc.fr/sor.htm. PMID- 12739333 TI - [Standards, Options and Recommendations for the use of PET-FDG in cancerology. Technical bases and regulatory principles]. PMID- 12739334 TI - [Standards, Options and Recommendations for the use of PET-FDG in cancerology. Results in bronchopulmonary and pleural cancers]. PMID- 12739335 TI - [Standards, Options and Recommendations for the use of PET-FDG in cancerology. Results in skin and ocular melanoma]. PMID- 12739336 TI - [Standards, Options and Recommendations for the use of PET-FDG in cancerology. Results in gynecological neoplasms]. PMID- 12739337 TI - [Standards, Options and Recommendations for the use of PET-FDG in cancerology. Results in digestive system neoplasms]. PMID- 12739338 TI - [Standards, Options and Recommendations for the use of PET-FDG in cancerology. Results in cancer of the upper aerodigestive tract]. PMID- 12739339 TI - [Standards, Options and Recommendations for the use of PET-FDG in cancerology. Results in urological cancers]. PMID- 12739340 TI - [Standards, Options and Recommendations for the use of PET-FDG in cancerology. Results in the treatment of Hodgkin's disease and malignant non-Hodgkin's lymphoma]. PMID- 12739341 TI - [Standards, Options and Recommendations for the use of PET-FDG in cancerology. Results in sarcomas]. PMID- 12739342 TI - [Standards, Options and Recommendations for the use of PET-FDG in cancerology. Results in unknown primary neoplasms]. PMID- 12739344 TI - The future belongs to the committed, the dedicated, & the caring. PMID- 12739343 TI - [Standards, Options and Recommendations for the use of PET-FDG in cancerology. Results in cancer of the thyroid]. PMID- 12739345 TI - Living longer, living better: the promise of aging research. AB - With the increasing numbers of the population aging, we are paying close attention to the opportunities and to the challenges of growing older. The needs of an aging populace are being met with a dramatic increase in research into the biology of aging, reducing disease and disability, and behavioral and cognitive aspects of aging. PMID- 12739346 TI - Alzheimer's disease. Hope for the future. AB - It has been only two decades since researchers discovered that most "senile dementia" is the result of a specific disease process, Alzheimer's disease (AD), and not an inevitable consequence of aging. In these 20 years, scientists have accumulated an extraordinary body of research. Until recently, preventing or curing AD was considered, at best, a distant possibility. Today, the picture is considerably brighter. PMID- 12739347 TI - Better care for our aging. AB - All physicians need to receive basic education and training in the care of older people during their medical studies to ensure adequate and appropriate health care of our aging population. The benefits of this will be felt across the entire health care spectrum, including home care and hospice. PMID- 12739348 TI - The three-legged stool of home care technology. Breakthroughs today, tomorrow, & in the future. AB - For a home care technology--or any technology for that matter--to achieve "breakthrough" status it must have three fundamental attributes. Without one of the three attributes, like a three-legged stool, the technology will not stand. PMID- 12739349 TI - Nutrition in disease prevention & treatment. AB - More focus is being placed on the role of nutrients in preventing and forestalling progression of chronic disease. Nutrition science continues to identify nutrients and patterns that are essential for healthy aging as well as for prevention and treatment of disease. PMID- 12739350 TI - Is there a doctor in the house? AB - The Washington Hospital Center initiated a geriatric medical and house call service for its local elderly population. Preliminary data shows that ER visits have been reduced by about 30 percent and admissions reduced by 10 percent, compared to the same patients in the year before enrollment. In most cases, the medical care is more effective, less costly, and more accessible for both the family and the health care system. PMID- 12739351 TI - Infection control in the home. PMID- 12739352 TI - Trends and treatment of cerebral aneurysms: review and case studies. AB - Subarachnoid hemorrhage (SAH) often results in devastating neurological deficits requiring hospitalization and loss of independence. This is often a difficult time for patients and their families who are struggling to cope with this sudden illness. Current treatment options include surgical clipping of the aneurysm or endovascular obliteration using Guglielmi Detachable Coils. Our purpose in writing this paper will be to review the factors that determine the choice of treatment, and to discuss how nurses can provide our patients and families with the support and teaching needed during this difficult time. PMID- 12739353 TI - Deciding on surgery: supporting parents of infants with craniosynostosis. AB - Families face a difficult decision when choosing whether or not to have surgery for their infant with isolated craniosynostosis. While the skull deformity may not affect the child's physical health, growth, or development, it may have psychosocial impacts on a child's social-emotional development, self-esteem, and interpersonal interactions. Parents are challenged to balance surgical risks with potentially positive effects on their child's psychosocial health. The purpose of this research study was to explore parental decision-making related to surgery for isolated craniosynostosis, and to identify strategies that facilitate their decision-making. A thematic content analysis of focus group interview data revealed four themes that encapsulated the process of parental decision-making. The decision for some parents was agonizing. However, others found the decision relatively straightforward. In all cases, parents spent time thinking and gathering information. Certain critical events helped parents make their decision. Parents identified a number of strategies that would be helpful to the decision-making process. PMID- 12739354 TI - [Resected cases with primary lung cancer after preoperative high dose irradiation]. AB - After greater than 60 gray (Gy) irradiation, we performed the pulmonary resection in the 18 primary lung cancer cases. The mean irradiation dose to the tumor was 68.2 (range 60-101) Gy, and the mean irradiation dose to the bronchial stump was 47.1 (range 0-82) Gy. Median time from end of irradiation to surgical resection was 136 (range 20-894) days. One partial resection, 9 single lobectomies, 3 double lobectomies, and 5 pneumonectomies were done. Mainly, we closed the bronchial stump by the automatic stapling device and additional hand suturing. The bronchial stump was covered in the 12 cases by the owner stalk thymus, the intercostals muscular flap, the omentum flap, and et al. The major postoperative complications due to preoperative irradiation were not seen. Bronchopleural fistulas did not occur. Pathologically, the wall of the submucosal capillary vessels were getting thick in the patients operated more than 3 months later after irradiation. In such cases with the decrease of the blood flow, the bronchial stump should be covered. The pulmonary resection after the high dose irradiation was considered to be tolerable. PMID- 12739355 TI - [Acute massive pulmonary embolism occurring during orthopedic surgery]. AB - We present an emergent surgical case of massive pulmonary embolism (MPE) that happened during orthopedic surgery. A 38-year-old man, who had bone fractures with his lumbar vertebra and ankle, underwent the internal fixation of the tibial bone with tourniquet under general anesthesia in our hospital. During this surgery, the pulse oxymeter showed a drop of arterial oxygen saturation suddenly. Immediately we installed a transesophageal echo (TEE) probe in the patient, and detected enlargement of the right ventricle and right atrial thrombus. Half an hour later, the thrombus disappeared from the right atrium and the patient showed hemodynamic shock. We performed emergent embolectomy immediately under moderate hypothermic complete cardiopulmonary bypass. The postoperative course was uneventful, and one month later, his orthopedic surgery underwent completely. We conclude that TEE was a useful devise for the diagnosis of intraoperative MPE. PMID- 12739356 TI - [Off-pump heart surgery for the removal of a ruptured, detached, and entrapped cutting balloon]. AB - A 84-year-old female with a ruptured, detached, and entrapped cutting balloon was successfully undergone the surgical removal under beating heart. But the left anterior descending coronary artery (LAD) was severely dissected by ruptured cutting balloon. So we had to anastomose left internal thoracic artery (LITA) to the distal part of LAD. Postoperative coronary angiography showed the narrowing of LAD and the effective revascularization was not obtained. We must recognize that the intima of the coronary artery is impaired by the ruptured cutting balloon more severely than we expected. PMID- 12739358 TI - [The effect of intra-aortic balloon pumping on the graft of coronary artery bypass grafting]. AB - The effect of intra-aortic balloon pumping (IABP) on the graft of coronary artery bypass grafting (CABG) was examined. These patients needed IABP when they were weaned from cardiopulmonary bypass. The flow wave pattern in the graft was acquired and analyze by the slope, peak flow rate and area of systolic phase and diastolic phase. The flow wave pattern different between graft to a left anterior branch and the graft to the right coronary artery. IABP was significantly effective for the flow to the left anterior branch. However, the effect of IABP on the graft flow to the right coronary artery was not significant. In the natural heart, the flow rate of the left coronary artery predominates during the diastolic phase. However this predominance is not clear in the right coronary artery. Our results suggested that this phenomenon in the natural state caused a difference the flow wave pattern between graft of left coronary artery and that of the right coronary artery. PMID- 12739357 TI - [Superior results of ketone body ratio in pulsatile normothermic cardiopulmonary bypass; comparison with non-pulsatile cardiopulmonary bypass]. AB - Sixteen patients undergoing aortocoronary bypass surgery under normothermic cardiopulmonary bypass were divided into 2 groups according to the either addition or none of pulsatility induced by intra-aortic balloon pumping (IABP). In those patients, hepatic blood flow was measured 3 times before, during and after cardiopulmonary bypass. Additionally, arterial and hepatic ketone body ratios [(AKBR) and (HKBR)], and hepatic venous saturation (ShvO2) were measured throughout and after the surgery. RESULTS: The hepatic blood flows measured at 3 different times at the surgery were much more in the pulsatile group (p < 0.05). The values of AKBR, indicator of mitochondrial redox potential in hepatocytes, were maintained in nearly normal in the pulsatile group, but were suppressed in the non-pulsatile group. This trend was much more obvious in the values of HKBR. The significantly lower ShvO2 levels were observed in the non-pulsatile group during the cardiopulmonary bypass (p < 0.05). CONCLUSIONS: Pulsatile normothermic cardiopulmonary bypass induced by IABP provides better liver perfusion and results in a better hepatic metabolism than non-pulsatile cardiopulmonary bypass. PMID- 12739359 TI - [Aortic valve replacement in a renal transplant recipient]. AB - A 65-year-old male was admitted to our hospital for surgical treatment of congestive heart failure with aortic regurgitation. He had received renal transplantation 15 years before in the United States, and had been under immunosuppressive regimen with ciclosporin and mycophenolate mofetil. Although the renal allograft function had been gradually deteriorating, and preoperative serum creatinine level was 1.8 mg/dl, and it decreased to 1.5 mg/dl after aortic valve replacement. Cryopreserved aortic allograft was needed for the aortic valve replacement. The reasons are; the patient may need hemodialysis (HD) or retransplantation of the kidney in the future, and the immunosuppressive therapy for kidney will provide good immunologic environment for second allograft, i.e.- aortic valve. He tolerated the operation well and the immunosuppressive agents were continued in the perioperative period. He is now in New York Heart Association (NYHA) class I. PMID- 12739360 TI - [Perioperative management of patient with pulmonary operation associated with interstitial pneumonia]. AB - The prognosis of patients complicated with interstitial pneumonia (IP) who undergo pulmonary operations is very poor if acute exacerbation occurs after the operation. We have experienced 6 cases in which operations were performed on lungs complicated with IP The first patient died due to acute exacerbation, but the subsequent 5 patients had good postoperative courses due to 1) short-term administration of low-dose steroid before and after the operation, 2) avoidance of administration of high concentration of oxygen during and after the operation, and 3) administration of erythromycin before and after the operation, 4) N acetylcysteine inhalation and administration of dl-alpha-tocopherylnicotinate before and after the operation. It is thought that careful management of such patients is needed to prevent acute exacerbation even after discharge from hospital. PMID- 12739361 TI - [Experience of off-pump coronary artery bypass grafting with starfish heart positioner]. AB - Recent advances in off-pump coronary artery bypass (OPCAB) technique have demonstrated that multivessel coronary revascularization can be safely performed in selected patient without cardiopulmonary bypass. Numerous positioning technique and devices have been developed to assist access to coronary vessels during OPCAB, such as deep pericardial suture, surgical pads, and cardiac positioning device and stabilizers. We used new cardiac positioning device Starfish heart positioner (Medtronic Corporation) for 2 case of OPCAB. Starfish heart positioner was very useful to successfully perform multivessel coronary bypass on all walls of the beating heart. Starfish heart positioner will facilitate and expand the ability to easily and consistently perform OPCAB. PMID- 12739363 TI - [Congenital tracheal stenosis due to complete cartilage rings with right pulmonary agenesis]. AB - A 2-month-old male infant with severe dyspnea was diagnosed as having right pulmonary agenesis at birth and was admitted to our hospital after tracheal intubation with an endotracheal tube of 3 mm in diameter. However, the trachea was too stenotic to place the tube in the proper position. Chest X-ray on admission showed pneumonia of the left lung. Preoperative chest computed tomography (CT) scan and bronchoscopy showed that from the level of 12 mm beneath the coricoid cartilage, the trachea tapered and continuing to the tracheal carina and that the smallest tracheal level was located 18 mm distal from the coricoid cartilage, the area of which was 4 mm2. His respiratory condition rapidly deteriorated in spite of intravenous administration of antibiotics and mechanical ventilation. Percutaneous cardiopulmonary support (PCPS) was used to maintain his pulmonary function, and pericardial tracheoplasty was performed. Chest X-ray immediately after the operation did not show left lung reexpansion due to severe pulmonary edema. High-dose steroid pulse therapy was performed, but it was not effective. He died from acute respiratory failure due to infantile respiratory distress syndrome (IRDS) on postoperative day 3. The outcome in this case shows that it is very risky to repair tracheal stenosis in a patient with pneumonia using PCPS. PMID- 12739362 TI - [Strategy for reducing mortality in the procedure of distal arch aneurysm]. AB - We investigated various surgical procedure of distal arch aneurysm to propose a safer strategy with less mortality. From January 1998 to March 2001, we operated 10 cases of distal arch aneurysm. Different methods were applied over the years to reach the current improved technique, which is a combination of retro grade general perfusion, median sternotomy, deep hypothermic circulation arrest with selective cerebral perfusion, pharmacological cerebral protection, none touch aortic method, and open distal anastomosis. We have managed to improve postoperative complications, which we experienced many times in the past. We have had no post-operative cerebral accidents, except for 1 case of death by and mesenteric arterial obstruction due to leukemia. In conclusion, our method is safe and feasible and can expect the reduction of post-operative mortality. PMID- 12739364 TI - [Total arch replacement for right aortic arch with Kommerell diverticulum and aberrant left subclavian artery]. AB - A 57-year-old man suspected of having angina pectoris underwent coronary angiography and comprehensive examination, which revealed a right-side aortic arch accompanying Kommerell diverticulum and a aberrant left subclavian artery. Esophagography indicated that the esophagus was compressed on its right posterior side and the computed tomography (CT) revealed that the posterior side of the tracheal was compressed, however, the patient experienced no difficulty in breathing, hoarseness of voice or dysphasia. The size of the aortic diverticulum was less than 5 cm and the patient showed no symptom, however, if it was left untreated, there was a risk of rupture in the future. Also the esophagus and tracheal may develop complications due to prolonged compression. Therefore, we decided that the case required surgical operation. Total arch replacement was performed through mediastinotomy and right posterolateral in the 4th intercostal. The postoperative condition was good, and the patient was discharged without any complications. PMID- 12739365 TI - [An effective case of nitric oxide inhalation therapy in pulmonary hypertensive crisis in cardiopulmonary bypass following palliative operation for ventricular septal defect with pulmonary hypertension]. AB - A 4-month-old female with ventricular septal defect (VSD) and severe pulmonary hypertension (PH) underwent a patch closure for VSD. She could not be weaned from cardiopulmonary bypass (CPB) after the intracardiac repair due to PH crisis. Nitric oxide inhalation therapy during partial CPB enabled her to be weaned from CPB. This therapy could be gradually taped off and quit 7 days after the operation. Nitric oxide inhaled therapy is considered to be an excellent treatment for PH crisis during CPB in congenital cardiac surgery. The causes of PH crisis were also discussed in reference to the date of endothelin-1 (ET-1) measured during the operation. PMID- 12739367 TI - [Traumatic diaphragmatic hernia repaired by video-assisted thoracic surgery; report of two cases]. AB - Two cases of traumatic diaphragmatic hernia treated by video-assisted thoracic surgery (VATS) were experienced. Both a 67-year-old female (left side) and a 44 year-old female (right side) was admitted to our hospital in a traffic accident. These patients were diagnosed as diaphragmatic hernia by chest magnetic resonance imaging (MRI). We tried VATS to this trauma, but the combined mini-thoracotomy for 7 cm in length was necessary to replace sliding organs for both cases. The rupture of diaphragm was repaired by direct suture of 2-0 viclyl. VATS will be recommended in the hernia of the diaphragm. PMID- 12739366 TI - [Aortic valve replacement with a freestyle stentless valve using the modified subcoronary technique and hemiarch replacement for bicuspid aortic valve and ascending aortic aneurysma; report of a case]. AB - An association between bicuspid aortic valve disease and ascending aortic aneurysma has long been recognized. Root replacement with a composite valve graft for such disease is a well-established technique. But it may involve serious technical difficulties, and may be a more time-consuming procedure than separate valve replacement and graft replacement. We performed an aortic valve replacement with Freestyle stentless valve using the modified subcoronary technique and hemiarch replacement for a 72-year-old man with severe aortic stenosis and ascending aortic aneurysma. Angiographic studies after surgery showed no residual aortic regurgitation (AR) and no deformity of aorta. This technique is an acceptable option for an aortic disease and ascending aneurysma in elderly patients. PMID- 12739368 TI - [Thoracoscopic resection of localized fibrous tumor of the pleura; report of a case and review of the Japanese literature]. AB - A 61-year-old female was admitted with abnormal shadow on chest X-ray. Chest computed tomography (CT) showed a clearly localized flat tumor at the peripheral region of the left lung. We suspected localized fibrous tumor of the pleura (LFTP) and performed video-assisted thoracic surgery (VATS). Under the thoracoscopy, the tumor was connected to the visceral pleura with a short pedicle and completely resected with safety margin. Pathological findings, including immunohistochemical examination, definitely revealed benign LFTP. VATS is a good indication for the diagnosis and treatment of LFTP. This case was reported together with some reviews of the literature. PMID- 12739369 TI - [Subdiaphragmatico-mediastinal lipoma through the foramen of Bochdalek; report of a surgical case]. AB - A 53-year-old man was admitted to our hospital for evaluation of chest abnormal shadow at the left cardiophrenic angle. Computed tomography (CT) scan revealed a homogeneous mass of fatty density on the left diaphragm. Magnetic resonance imaging (MRI) demonstrated a well-demarcated tumor mass with a fatty signal intensity and, in the sagittal view, the mass showed continuity into the retroperitoneal fatty tissue. Extirpation of the tumor was performed under thoracoscopic procedure. The tumor was adhesive closely to diaphragm and extended into the retroperitoneal space through the foramen of Bochdalek. Pathologically the tumor was diagnosed as a mature lipoma. This tumor was considered to originated from the retroperitoneal fatty tissue under the diaphragm and to herniate into the mediastinum through the foramen of Bochdalek. PMID- 12739370 TI - [Intrathoracic chronic expanding hematoma; report of a case]. AB - A case of a patient with a intrathoracic chronic expanding hematoma presenting as a neoplasm is reported. A chronic encapsulated intrathoracic hematoma is rare condition and is clinically included in a category of chronic hemorrhagic empyemas as a type of chronic empyemas in Japan. The clinical picture of our case suggested a slowly growing tumor. Though needle biopsy was performed, only blood was aspirated. Intraoperative findings revealed a intrathoracic hematoma with a tough capsule. It adhered to the chest wall hardly. Histopathological investigations revealed that the capsule consisted of a collagenous outer layer and a newly vascularized inner layer with fibrosis. Hemosiderin deposits suggested recurrent bleeding from fragile vessels. The mechanisms of hematoma formation and its expanding nature are discussed. The hematoma appeared to have been enlarged by repeated exudation or bleeding from capillaries inside the capsule. PMID- 12739371 TI - [Antimicrobial susceptibility tests and resistant strain of Helicobacter pylori]. AB - The antimicrobial susceptibility test was necessary for the eradication therapy of Helicobacter pylori infections. This is because, clarithromycin resistant strains has became an increasing problem. In this study, we used the antimicrobial susceptibility test which was compare with the agar gradient method, Etest, and broth microdilution method (dry plate) with 4 antimicrobial agents. The results strongly suggested that broth microdilution method was the best method in order to test the antimicrobial susceptibility of H. pylori. On the other hand, 393 H. pylori stains isolated during 1994-1998 from clinical patients were tested for antimicrobial susceptibility test to amoxicillin, clarithromycin, metronidazole, and minomycin. There were no resistant strains to amoxicillin and minomycin. But clarithromycin and Metronidazole resistant strains were recognized in 85 (22.0%) and 36 (21.7%) strains. PMID- 12739372 TI - [Increasing drug resistance in Vibrio cholerae O1 and non-O1 strains isolated from diarrheal cases in Japan]. AB - Drug resistance trends were investigated for 271 Vibrio cholerae O1 (V.c O1) and 401 V. cholerae non-O1 (V.c non-O1) strains isolated from mainly imported diarrheal cases during 1981-2001 in Japan. The results of drug resistance test using 8 drugs (CP, TC, SM, KM, ABPC, ST, NA, and NFLX) showed that 34.7% of the V. c O1 strains and 15.7% of V.c non-O1 strains were multi-drug or mono-drug resistant. The incidence of drug resistant strains has increased since 1991, and it has been remarkable in V.c O1 strains that increased from 1.2% in 1981-1985 to 70.8% in 1996-2001. The drug resistance patterns of the resistant strains classified into 6 types in V.c O1 and 21 types in V.c non-O1. The prevalent patterns recognized were SM (75.5%), CP.TC.SM.ST (10.6%) and CP.SM.ST (8.5%) in V.c O1, and SM (25.4%) and ABPC (25.4%) in V.c non-O1. Ten V.c O1 strains (3.7%) and 10 V.c non-O1 strains (2.5%) were multi-drug resistant including TC. Among those, 13 strains were isolated from travelers who returned to Japan from Thailand. One V.c O1 strain (0.4%) and 6 V.c non-O1 strains (1.5%) were NA high resistant and fluoroquinolones low-sensitive. Among those, 4 strains were isolated from travelers who returned to Japan from India. PMID- 12739373 TI - [Analysis of tuberculin skin test in employees of Nagasaki University Hospital]. AB - The tuberculin skin test (TST) was conducted in 1,087 employees of Nagasaki University Hospital. The size of erythema (T1) was 27.3 +/- 17.0 mm, and 8.6% of all participants showed < = 9 mm, 49.9% showed 10-29 mm, 41.5% showed > = 30 mm. Laboratory technologists showed significantly larger reactions than other groups. Doctors and nurses working on the tuberculosis ward showed larger reactions than those working on the non-tuberculosis wards. Reactions tended to be larger with aging, and the younger employees in their 20s showed significantly smaller reactions than those in their 40s. The second TST was carried out in 253 employees whose T1 was below 30 mm. The size of erythema enlarged from 16.1 +/- 6.6 mm (T1) to 26.4 +/- 15.4 mm (T2), with the difference (booster phenomenon) of 10.2 +/- 14.4 mm. T2 and T2-T1 were larger in nurses working on the tuberculosis ward than those on the non-tuberculosis wards. It was suggested that laboratory technologists, nurses and doctors especially working on the tuberculosis ward are at risk of tuberculosis infection, and the two-step TST was an essential means to know the baseline reactivity, and to determine recent tuberculosis infection. PMID- 12739374 TI - [Targeted bacteremia surveillance throughout a year--comparison of community acquired and hospital-acquired infection]. AB - To keep an eye on severe nosocomial infection and to evaluate the clinical difference of blood-stream infection between community-acquired and hospital acquired infection, a survey of blood culture was performed in National Tokyo Medical Center from the period between November 2000 and October 2001. There were 252 episodes detected in 219 patients (80 community-acquired episodes in 80 patients and 172 hospital-acquired episodes in 139 patients). The three most common foci of infection/pathogens were as follows: in the community-acquired cases; urinary tract, pneumonia, infective endocarditis/Escherichia coli, viridant group of streptococci, Streptococcus pneumoniae, and in the hospital acquired cases; intra-venous catheter, urinary tract, neutropenia-related bacteremia/Staphylococcus aureus, coagulase negative Staphylococcus, Enterococcus. Fifteen patients with community-acquired bacteremia and 37 patients with hospital-acquired bacteremia had been died within a month of the episode; the mortality was not significantly different between the both. The average of peak serum concentrations of C-reactive protein during the episodes of community acquired bacteremia was higher than that of hospital-acquired bacteremia. These findings probably show that life threatening bloodstream infections seemed to be more common in the community. The rate of nosocomial bacteremia was approximately 1%, and no outbreak was observed during the period. Targeted bacteremia surveillance is maybe useful and efficient method to detect severe hospital acquired infections. PMID- 12739375 TI - [Clinical evaluation of performance of a new diagnostic method for deep mycosis by measuring beta-glucan concentration in the blood]. AB - beta-Glucan Test MARUHA for high sensitive diagnosis of deep mycosis which was developed recently detects (1-->3)-beta-D-glucan, a component of the cell wall. The performance of beta-Glucan Test MARUHA is evaluated in this report. Although existing methods to detect (1-->3)-beta-D-glucan have trouble with sulfa drugs, hemolysis, and immunoglobulin G (IgG), these problems were overcome by the beta Glucan Test MARUHA: No effect was observed for beta-Glucan Test MARUHA at lower than 200 micrograms/ml, 500 mg/dl, and 6,000 mg/dl of sulfa drugs, hemoglobin, and IgG, respectively. The effect of drugs administrated on the measurement value of beta-glucan Test MARUHA was checked at several concentrations. However, almost no effect of drugs, such as, 5 kinds of antifungals, 8 kinds of antibiotics, a kind of antibacterial, 2 kinds of infusions, and a kind of contrast media was observed at the practical concentrations. PMID- 12739376 TI - [Comparison of four diagnostic methods using clinical blood by measuring (1-->3) beta-D-glucan]. AB - Using Amebocyte lysate of horseshoe crab to measure (1-->3)-beta-D-glucan specifically, a component of the cell wall, several kinds of diagnostic methods for deep mycosis are in practical use in Japan. However, the most important problem is that the judgment of positive or negative is method dependent. To elucidate the cause of the problem, each measurement value of the identical sample by four methods, beta-Glucan Test Maruha (MARUHA), beta-Glucan Test Wako (WAKO). FUNGITEC G Test (FUNGITEC-G) and FUNGITEC G Test MK (FUNGITEC-MK) was compared with the clinical data using 119 cases and 289 tests. Each case was divided into three groups; proven fungal infection, probable fungal infection and non-fungal infection. The negative cases for all the methods tested in the groups of proven fungal infection and probable fungal infection were allergic bronchopulmonary aspergillosis and cryptococcosis, and that for all the methods tested except FUNGITEC-MK method in the group was pulmonary aspergilloma. It seems that these cases cannot be detected correctly by only measuring (1-->3) beta-D-glucan. On the other hand, the ratio of false positive, positive for non fungal infection group was high in the case of FUNGITEC-MK. About 23% against the total case was positive for FUNGITEC-MK method, but negative for MARUHA, WAKO, and FUNGITEC-G methods. Although the difference of the sensitivity among four methods was not observed, the specificity, the diagnostic efficiency, and the positive predictive value of FUNGITEC-MK method were remarkably lower than those of the other methods due to false positive measurement. In conclusion, MARUHA, WAKO and FUNGITEC-G except FUNGITEC-MK is not method dependent. It is suggested that FUNGITEC-MK detects non-specific reaction resulted in false positive. PMID- 12739377 TI - [Aeromonas septicemia complicated with liver cirrhosis in Tokyo Women's Medical University]. PMID- 12739378 TI - [Respiratory infectious complications after acute ischemic stroke]. AB - BACKGROUND AND PURPOSE: Respiratory infection is a frequent complication in acute ischemic stroke, but it seems to have been made light of in stroke care. The purpose of this study is to examine the clinical characteristics of respiratory infectious complications in patients with acute ischemic stroke. METHOD: Two hundred and fifty-eight consecutive patients (158 men, 100 women, 70.6 +/- 12.9 years old) with acute ischemic stroke were admitted to our hospitals between May and October in 1999. Age, gender, history of stroke, the severity of stroke on admission, stroke subtype (lacunar brain infarction, atherothrombotic brain infarction, cardioembolic brain infarction, and others), aspiration, naso-gastric tube feeding, vascular risk factors, the length of hospital stay and outcome of patients were noted. We compared them between patients with and without respiratory infections. RESULTS: Forty-five (17.4%) patients were developed respiratory infections. Cardioembolic stroke patients were more frequently developed respiratory infections (67%) compared with other stroke subtypes. The independent risk factors for respiratory infectious complications by multiple logistic regression model were the aspiration (OR, 5.513; 95% CI, 1.793-16.946) and the severity of stroke on admission (OR, 1.090; 95% CI, 1.034-1.150). Mortality of patients with respiratory infectious complications was as high as 24%, and all survivors discharged to another hospital. After adjustment for age and the severity of stroke, respiratory infection was one of the independent risk factors of poor stroke outcome (OR, 5.838; 95% CI, 1.792-19.018). CONCLUSION: Aspiration and the severity of stroke independently predict development of respiratory infectious complication in acute ischemic stroke. Respiratory infections may make worse their stroke outcome. A measure to infectious complications and aspiration needs to be taken for the patients suffering from severe ischemic stroke. PMID- 12739379 TI - [Non-persistent "doll phenomenon" in a patient with right thalamic infarction]. AB - An 81-year-old right-handed woman was admitted because of acute dysarthria and left hemiparesis. She had lived herself without aids until the admission. On neurological examination she was confused and disoriented. She was ambulant, but had mild dysarthria and mild left hemiparesis. Neuropsychological tests showed severe impairment of memory, mild impairment of visual cognition, decreased fluency of word recall and mild paramnesia, but no acalculia, agraphia, aphasia or apraxia. MRI of the brain showed small infarction in the right anterior thalamus. 123I-IMP SPECT demonstrated a decrease in CBF of the thalamus, basal ganglia and frontal lobe on the right. During admission, she always played with a doll as if she took it as a real baby. This peculiar symptom. "doll phenomenon" continued for approximately three months later. The "doll phenomenon" usually appears in demented patients with diffuse mental deterioration or dysfunction of the frontal lobe. The present patient had not been demented until the onset of the thalamic infarction, and disturbance of cognition caused by the right thalamic infarction probably produced the "doll phenomenon". PMID- 12739380 TI - [Compulsive repetition of movements in a case of progressive supranuclear palsy]. AB - A 51-year-old right-handed man with an 8-year history of progressive supranuclear palsy developed a peculiar behavior including compulsive repetition of movements. At the age of 47, it was noted that he continuously scratched his trunk with the right hand. He also scratched his thighs until his trousers were torn. On the scalp, posterior neck and forearms, patches of skin were scraped away and the bleeding wounds were scratched over and over again. Although he perceived pain, he could not terminate the strange scratching movement. He could not stop his eating action even after the bowl became empty. He could not leave the bathroom because he continued to wipe the anus after defecation. Yet the patient was not demented. Psychometric assessments including Wechsler adult intelligence scale revised, meaningful and meaningless syllables memory test. Raven's color matrix test, Wisconsin card sorting test (Keio version), and modified Stroop test were within the normal range. When requested to write a letter or a figure, he was always successful, however, he endlessly wrote the same letter or figure until he was told to stop. Although he repeated the behavior and could not terminate the action by himself, he immediately stopped the action on verbal command. When another task was given during the repetitive action, he responded quickly and successfully. His strange behavior was compulsive because he wanted to cease it, but it was not compulsive-obsessive behavior because he did not feel anxiety or pain when his repetitive action was stopped. It was different from stereotypy because he repeated not only meaningless movements but also meaningful complex actions. Unlike perseveration, he changed his activity immediately and successfully when a new task was given. Magnetic resonance imaging findings were remarkable only for mild atrophy of the midbrain tegmentum. Single photon emission computed tomography with ethyl cysteinate dimer demonstrated hypoperfusion in the frontal lobes and the left thalamus. A thalamofrontal disconnection or dysfunction is suggested as the mechanism underlying the compulsive repetifim of moving its based on the SPECT findings in this patient. PMID- 12739381 TI - [A case of cavernous sinus cavernous hemangioma presenting with cavernous sinus syndrome]. AB - The authors report an unusual case of a 50-year-old woman presenting with cavernous sinus syndrome, who had a cavernous sinus cavernous hemangioma (CSCH). The acute onset of her symptoms including pain of the right eye, blephaloptosis of the right eye, diplopia, and sensory disturbance of the right face was similar to those of Tolosa-Hunt syndrome. Magnetic resonance imaging and angiography showed a tumor in the right cavernous sinus. Although she showed improvement of the symptoms after receiving oral corticosteroids, follow-up neuroradiological investigations after a year from the onset revealed the mass in the right cavernous sinus which grew up in size. The histopathological findings obtained from the biopsy of the mass demonstrated a CSCH. Our findings suggest that the growth mechanism of CSCH could be progressive ectasia of vessels or their autonomous development at the edges of the lesions. PMID- 12739382 TI - [A case of pure anarithmetia associated with disability in processing of abstract spatial relationship]. AB - A 35-year-old right handed man developed pure anarithmetia after an left parieto occipital subcortical hemorrhage. His intelligence, memory, language, and construction ability were all within normal limits. No hemispatial neglect, agraphia, finger agnosia, or right-left disorientation were noted. He showed no impairments in reading numbers aloud, pointing to written numbers, writing numbers to dictation, decomposition of numbers, estimation of numbers of dots, reading and writing of arithmetic signs, comprehension of arithmetic signs, appreciation of number values, appreciation of dots' number, counting aloud, alignment numbers, comprehension of the commulative law and the distributive law, retrieval of the table value (ku-ku), immediate memory for arithmetic problems, and use of electric calculator. He showed, however, remarkable difficulty even in addition and subtraction between one figure digits, and used counting on his fingers or intuitive strategy to solve the problems even when he could solve them. He could not execute multiplication and division, if the problems required other than the table value (ku-ku). Thus, he seemed to have difficulties in both of elemental arithmetic facts and calculating procedures. In addition, his backward digit span and reading of analogue clocks were deteriorated, and he showed logico-grammatical disorder of Luria. Our case supports the notion that there is a neural system which was shared in part between processing of abstract spatial relationship and calculation. PMID- 12739383 TI - [A case of bilateral perisylvian polymicrogyria with epileptic attacks of focal inhibitory seizure followed by complex partial seizure]. AB - A 20 year-old woman began to have epileptic attacks of focal inhibitory seizure with paralysis and hypesthesia of her left or right upper limb followed by complex partial seizure several times a week since age 19. She was born by breech presentation and umbilical cord was coiling around her neck at birth. EEG showed spike foci on P 3, O1 and T5. Cerebral MRI with 4 mm-section inversion recovery image revealed bilaterally symmetrical polymicrogyria involving the posterior portion of sylvian fissure extending posteriorly to the inner cortex of the postcentral gyrus and the supramarginal gyrus, and she was diagnosed as bilateral perisylvian polymicrogyria. 99mTc-ECD SPECT showed increased cerebral blood flow over the bilateral polymicrogyric lesion. On cerebral MRA, bilateral middle cerebral arteries were narrow all way through. Epileptic attacks were controlled with zonisamide and carbamazepin. This is a rare case of bilateral perisylvian polymicrogyria because epileptic attacks were the only manifestation and the patient showed neither mental retardation nor neurological abnormality. PMID- 12739384 TI - [Primary Sjogren's syndrome presenting with choreo-athetosis]. AB - A 69-year-old man presenting with choreo-athetosis was proved to have primary Sjogren's syndrome. Choreo-athetosis suddenly appeared in the bilateral legs and the right arm; it was predominant in the right limbs. On the neurological examinations, there was no abnormal finding except for this involuntary movement. Brain MRI showed no abnormal finding. 18F-Fluorodeoxyglucose PET revealed that glucose metabolism was relatively increased in the bilateral striatum and thalamus (left side dominant). Dry eye and thirst were present for 4 years, and a diagnosis of primary Sjogren syndrome was confirmed by ophthalmological examinations, lip biopsy and sialography. Choreo-athetosis disappeared after thioridazine hydrochloride was started, while steroid was not effective. PMID- 12739385 TI - [Reversible posterior leukoencephalopathy syndrome in two patients with microscopic polyarteritis nodosa]. AB - We report here two cases (72-year-old woman, 77-year-old-woman) with perinuclear anti-neutrophil antibody (P-ANCA) positive microscopic polyarteritis nodosa (mPN). Both patients presented with generalized convulsion, consciousness disturbance and severe hypertension several days after the administration of high dose methylpredonisolone followed by oral predonisolone. CT brain scan showed hypodensity area on bilateral posterior lobes. MRI T2 weighted image and FLAIR image showed increased signal intensity on the occipital gray and white matter. Although diffusion weighted MRI disclosed slightly high signal intensity, apparent diffusion coefficient (ADC) value at the occipital gray and white matter remained normal or even elevated. These findings, which were consistent with vasogenic edema, lead us to diagnose them as suffering from reversible posterior leukoencephalopathy syndrome (RPLS). After the second administration of high dose methylpredonisolone, their symptoms and signs, together with the MRI findings, gradually improved. To our knowledge, this is the first report concerning RPLS in a patient with mPN. PMID- 12739386 TI - [A case of pseudomigraine with pleocytosis]. AB - We report a case of pseudomigraine with pleocytosis (PMP) characterized by temporary neurological deficits and elevated cell counts in cerebrospinal fluid (CSF). A 28-year-old woman was admitted to our hospital with a second episode of right side throbbing headache accompanied by hemianopsia without scintillating scotoma of left side, hand numbness and weakness of left hand. Two months before the admission, she experienced a first identical episode, which lasted several hours. On admission to our hospital, neurological examination showed left hemianopsia, mild left hemiparesis, dysesthesia of left hand, exceeded tendon reflex of left upper limb, stiff-neck and positive Kerning's sign. CSF examination showed mild elevation of mononuclear cell counts. No abnormal findings on brain CT and MRI (including diffusion weighted image) were observed. 99mTc-HMPAO single photon emission computed tomography (SPECT) demonstrated extensive hypoperfusion at right cerebral hemisphere, corresponding to her neurological deficits. Her electroencephalography (EEG) showed reduced amplitude on the right occipital area. The reduced amplitude of cortical component of somatosensory evoked potential (SEP) by left median nerve stimulation were observed. On the third day after the admission, her symptoms improved and cell count of CSF was normalized. One week after the onset her SEP, EEG and SPECT were normalized on their retrials. She has never recurred these symptoms. We established a diagnosed of psedomigraine with pleocytosis as the first Japanese case. PMID- 12739387 TI - [A case of hemodynamic brain infarction with postprandial hypotension]. AB - A 76-year-old man with left internal carotid artery occlusion developed a progressing right hemiparesis. Brain MRI presented reinfarctions in the left anterior border zone and terminal zone in the left deep white matter. Ambulatory blood pressure monitoring showed a decrease in systolic blood pressure by more than 20 mmHg one hour after starting meals, which is considered as postprandial hypotension. The recurrent stroke occurred probably by a hemodynamic mechanism with the presence of internal carotid artery occlusion and postprandial hypotension. Administration of voglibose, an alpha-glucosidase inhibitor, improved postprandial hypotension. In patients with severe carotid or intracranial artery disease, the postprandial hypotension should be carefully monitored for prevention of hemodynamic brain ischemia. PMID- 12739389 TI - [A case of herpes simplex encephalitis with noticeable findings of SPECT]. AB - A 51-year-old man was admitted to our hospital because of fever, general fatigue and disturbance of consciousness. Neurological findings included disturbed consciousness, stiff neck and positive Kernig's sign. He was diagnosed as having herpes simplex encephalitis with HSV-DNA in the cerebrospinal fluid. MRI showed a lesion with low signal intensity in T1-weighted image and high signal intensity in T2-weighted image in the right temporal lobe. The single photon emission CT (SPECT) study showed discordance of 99mTc ethyl cysteinate dimer-SPECT (ECD SPECT) and N-isopropyl-p-[123I]-iodoamphetamine-SPECT (IMP-SPECT). Decreased signal of ECD in the lesion where IMP uptake was increased could be due to decreased esterase activity. This report suggests that ECD-SPECT could fail to detect cerebral blood flow in the lesion with severe metabolic damage. PMID- 12739388 TI - [A case of motor neuron disease with dementia and apraxia of the upper limbs]. AB - We report a 61-year-old, right-handed woman with motor neuron disease, dementia, and apraxia of the upper limbs. The patient developed clumsiness of the right hand and dysarthria two years and a half prior to admission. Neurological examination showed limb-kinetic apraxia and ideomotor apraxia, predominantly on the right side, in addition to dementia and anarthria. There was mild muscle wasting in the neck and hands. A muscle biopsy from the biceps muscle of arm as well as needle EMG revealed neurogenic changes compatible with motor neuron disease. Brain MRI indicated pyramidal tract degeneration. Three-dimensional brain perfusion imaging generated from SPECT demonstrated an asymmetric decrease in cerebral blood flow in the fronto-temporo-parietal regions, predominantly on the left side. This case suggests that asymmetric limb apraxia can be associated with motor neuron disease. PMID- 12739391 TI - [Functional neural reorganization following ischemic stroke in the precentral knob: an fMRI study]. AB - We evaluated a 63 year-old, right-handed woman by functional MRI (fMRI) in the early and the recovery stages following a cerebral infarction in the right precentral knob. An activated signal in the ipsilateral sensorimotor cortex and contralateral supplementary motor cortex during deteriorated hand grasping (left) was observed in the early stages, whereas being unable to detect in the functionally recovered period. When simultaneous bilateral hand grasping was performed, the activated signal around infarct region was enlarged in recovered period. The functionally neural reorganization processes relating recovered hand movement after localized cortical infarction (precentral knob) was suggested. PMID- 12739390 TI - [A young adult case of spontaneous basilar artery dissection]. AB - We reported a case of basilar artery dissection in a 20-year-old man suffering sudden onset of consciousness disturbance. Brain CT revealed a cerebral infarction of the whole territories of vertebro-basilar artery and his 3DCT showed the occlusion at the base of basilar artery. Autopsy revealed that the subintimal dissection was found at the base of basilar artery and the dissection was spreaded to the distal of bilateral posterior cerebral arteries. The characteristics of his vertebro-basilar artery were small in diameter, thin media and thickened intima. According to these findings, we supposed this rare case of basilar dissection occurred all at once based on a functional abnormality in his small vertebro-basilar arterial wall. PMID- 12739392 TI - [A case of transient global amnesia (TGA) showing bilateral hippocampal hypoperfusion by a new SPECT analyzing system, eZIS]. AB - We here reported a case of typical transient global amnesia (TGA). A 64-year-old right-handed man was suddenly unable to keep his recent memory without any event. There were no neurological deficits except for recent memory disturbances. He showed no causative findings of the attack on examinations of laboratory data, EEG and MRI. Examination of single photon emission CT (SPECT), using 99mTc-ECD, was performed in acute and chronic stages. We were able to detect bilateral hippocampal hypoperfusion in the acute stage using a new analyzing system, easy Z score Imaging System (eZIS). It showed extensive hypoperfusion also in the vertebrobasilar artery territory. Twenty-seven days later, these findings disappeared. Using this system, we are able to evaluate cerebral blood flow objective with anatomical information by SPECT. Therefore, we concluded that this analyzing system would be helpful to reveal the pathophysiological mechanism of TGA. PMID- 12739393 TI - [Secular increase in the incidence rate of drug-induced hepatitis due to anti tuberculosis chemotherapy including isoniazid and rifampicin]. AB - To investigate the secular change in the incidence rate of drug-induced hepatitis (DIH) due to anti-tuberculosis chemotherapy including isoniazid (INH) and rifampicin (RFP), but not including pyrazinamide (PZA), we retrospectively studied the incidence rates of DIH in patients treated with chemotherapy including INH and RFP in four periods 1980-83, 87-88, 91-92, and 1998-2000. The criteria for selection of the patients were as follows. 1. The serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured before, and one month (20-40 days) and 2 months (45-75 days) after starting anti tuberculosis chemotherapy. When the serum AST and ALT were measured twice or more during period 20-40 days or 45-75 days after starting anti-tuberculosis chemotherapy, the data obtained nearest to 30 or 60 days after were chosen as those of one or two months after starting chemotherapy, respectively. 2. The serum AST and ALT were within normal range before starting anti-tuberculosis chemotherapy. The normal range of serum AST and ALT were < or = 40 K-A and < or = 35 K-A (in 1980-83) or < or = 31 IU/l and 34 IU/l (in 1987-2000), respectively. 3. Chronic active hepatitis and cirrhosis patients were excluded. 4. All alive after completion of anti-tuberculosis chemotherapy. The numbers of the subjects who fulfilled the above criteria were 113, 135, 128 and 154 in 1980-83, 1987-88, 1991-92 and 1998-2000, respectively. DIH was defined serologically by serum AST > or = 40 K-A and/or ALT > or = 35 K-A (in 1980-83), or AST > or = 40 IU/l and/or ALT > or = 40 IU/l (1987-2000). The DIH incidence rate of the subjects classified by the year of treatment and age were examined, and the contributions of the risk factors for DIH, such as age, sex, alcoholics, previous liver disease history, HBs ag positivity, anti-HCV ab positivity, and hypoalbuminenia were studied, and none except the age over 80 y.o. was found to be a risk factor to DIH, in our subjects. In patients with the age over 80 y.o., daily doses of antituberculosis drugs RFP, INH and ethambutol (EB) were significantly higher in patients with DIH than those without DIH, but body weight and serum albumin level were not significantly different between these two groups. There was no risk factor to DIH in our patients less than 80 y.o. and this could be explained by the above mentioned criteria of study patients selection. To exclude the age dependence of the incidence rate of DIH in our subjects, the incidence rates of DIH were calculated in patients less than 80 y.o. by the period of treatment, and they were 10/111 (9.0%), 23/131 (17.6%), 26/123 (21.1%) and 32/117 (27.4%) in 1980-83, 87-88, 91-92, and 1998-2000, respectively. The secular increase of the incidence rate of DIH was statistically significant (p = 0.01). It is quite clear that this secular increase was not at all attributable to the above-mentioned risk factors. It is suspected that human liver has become more easily affected with INH and RFP in recent years. It is suggested that the new chemical compounds present in our increasingly complicated human milieu give heavier burdens on human liver, weaken the liver function, and enhance the capacity of INH and RFP to cause DIH. PMID- 12739394 TI - [Predominant location of pulmonary parenchymal lesions of tuberculosis primary complex in infants aged less than one year]. AB - It is useful to know the distribution of pulmonary lesions in the diagnosis of tuberculosis on radiological examination. The aim of this study was to investigate if there is predominant lung segment or lobe for tuberculous lesions in infants aged less than one year using contrast enhanced CT. We studied 57 infants (40 boys, 17 girls) who were diagnosed as tuberculosis by isolation of Mycobacterium tuberculosis or combination of family contact, radiographic findings suggesting tuberculosis, and positive reaction of 5 mm or more induration to PPD tuberculin. All the infants had lesions in mediastinal and/or hilar lymphonodes, and 54 out of 57 infants had parenchymal lesions as well. In the study of the segmental predominance of tuberculous lesions, each infant had a share of 100 points. If an infant had a single focus, all the points were distributed to the corresponding segment. If he or she had multiple foci, the 100 points were equally divided into affected lung segments. There was no significant difference between right (3385 points/10 segments) and left (2005 points/8 segments) lungs. The points in upper lobes (2224 points/5 segments) were significantly higher than the combined points of middle and lingual (896 points/4 segments) and lower (2270 points/9 segments) lobes (p < 0.05). The points in posterior lung segments (2839 points/7 segments) were significantly higher than the combined points of middle (436 points/3 segments) and anterior (2115 points/8 segments) lung segments (p < 0.05). These results suggest that upper lobes and posterior segments are predominant parenchymal regions of tuberculosis among infants less than one year, although tuberculous lesions may locate in any lung segment. PMID- 12739395 TI - [Problems about the management of active pulmonary tuberculosis patients undergoing haemodialysis--our experiences and nation-wide questionnaire survey]. AB - End-stage renal failure patients on chronic dialysis are high risk groups of tuberculosis due to attenuated cellular immunity. Patients receiving haemodialysis stay prolonged time inside the health-care facilities, thereby increased risk of tuberculosis transmission if a patient has active disease. So management of active pulmonary tuberculosis undergoing haemodialysis is important, however, the number of hospitals which are capable of taking care of such patients is estimated to be few in Japan. METHODS: From August 1994 through July 2002, 1059 active pulmonary tuberculosis patients (mean age; 57 +/- 19, male/female = 773/286) were admitted to Nishi-Kobe Medical Center, a 500-bed teaching hospital. Out of them, patients undergoing haemodialysis were retrospectively studied to describe the clinical characteristics of such cases. Then we conducted a questionnaire survey regarding the management of active pulmonary tuberculosis patients undergoing haemodialysis for 86 self-governing bodies in Japan. RESULTS: (1) Clinical characteristics of active pulmonary tuberculosis undergoing haemodialysis. We encountered 14 cases (mean age; 65 +/- 11, male/female = 7/7) of pulmonary tuberculosis undergoing haemodialysis during 8 years. In addition to pulmonary involvement, 3 pleural, one knee joint and one lymph node involvement was detected. Primary renal disease included diabetic nephropathy (n = 3), chronic glomerulonephritis (n = 3), congenital anomaly (n = 1), and unknown (n = 7). Nine cases were referred to our hospital from health care facilities located out of city or prefecture. In five cases it took more than three months from the onset or detection of abnormal chest X-ray findings to the admission to our hospital. Five cases developed pulmonary tuberculosis within the first year after the initiation of dialysis. None of the patients had a past history of tuberculosis. Cavitary lesion on chest X-ray was observed in only one case. Triple antituberculosis therapy was used in 9 patients, and 4 antituberculosis drugs were used in 5 patients. Antituberculosis therapy was successfully done in all cases except two patients who died of apoplexy and cerebral infarction. (2) The nation-wide questionnaire survey. Of the 86 self governing bodies we mailed, 66 self-governing bodies replied. Of them, 31% reported that they have experienced difficulties in the management of active pulmonary tuberculosis patients undergoing haemodialysis, and 25% reported the lack of health-care facilities to take care of such cases in their territory. They have referred such patients to hospitals located in the nearby prefectures or they have recommended antituberculosis therapy visiting a local haemodialysis facility. CONCLUSION: There are sometimes difficulties to manage active pulmonary tuberculosis patients undergoing haemodialysis in Japan. Health-care facilities to take care of such patients should be arranged and the formation of the network is necessarily. PMID- 12739396 TI - [Comparison between relative analogy and homology of 16S rRNA partial sequences between Mycobacterium szulgai and Mycobacterium malmoense]. AB - A lot of nucleotide sequences of some genes, especially 16S rRNA gene, are registered in the public data-base. In order to identify clinical mycobacterial strains, 16S rRNA gene partial nucleotide sequences from 15 strains of Mycobacterium malmoense and 24 strains of Mycobacterium szulgai which are stored in the Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association were determined. Then homology of the sequences to the data of type strains submitted to public database were determined. Relative analogy to type strains by delta DDH was also measured. In the area of nucleotide position 51-588 corresponding accession number X52930 which is Mycobacterium malmoense type strain 16S rRNA gene sequence data, the homology of some partial sequences with Mycobacterium malmoense strains to data of accession number X52930 were lower than that with Mycobacterium szulgai type strain data, accession number X52926. In the area of nucleotide position 31-568 or nucleotide position 31-588, the homology of all nucleotide sequence data to collect species data of type strains were higher than the homology to another species data. Nucleotide position 38, 40, 47 and 49 might be differential nucleotides conserved between Mycobacterium malmoense strains and Mycobacterium szulgai one. These results suggest that homology of about 500 bp's 16S rRNA gene nucleotide sequence data may not be enough for differential identification. Nevertheless, database of RIDOM, Ribosomal differentiation of Medical Microorganisms, identified partial nucleotide sequences between nucleotide position 51-588 corresponding accession number X52930 correctly, though some were lower than 97% homology (data not shown). Therefore quality-controlled 16S rRNA gene nucleotide sequence database could be used for differential identification. PMID- 12739398 TI - [University as business investment. Money from trade and industry more and more important for Swedish medical research]. PMID- 12739397 TI - [A new method of sputum pre-treatment for PCR preparation]. AB - A rapid sputum processing method was developed for PCR. As a protocol prepared for commercial kit is too much simplified, PCR inhibitor is not completely removed from sputum. We developed a new semi-alkaline protease method to dissolve sputum. From April 2001 to August 2001, 261 sputum samples were treated with our new method. Twenty-one cases (8.0%) were PCR positive and all the results perfectly coincided with the results of the conventional culture method. PMID- 12739399 TI - [Anti-IgE therapy--possible way in severe nut allergy. The hidden allergen in candy is a danger for risk groups]. PMID- 12739400 TI - [It's difficult to introduce evidence-based medicine in everyday health care. Time pressure and insufficient education are the greatest obstacles]. PMID- 12739401 TI - [Decision support in drug prescribing. A tool which must be well-functioning in a complex clinical reality]. PMID- 12739402 TI - [Free radicals and ailing proteins--the culprits behind Parkinson disease?]. AB - Parkinson's disease is one of the most common neurodegenerative diseases, and affects approximately 1% of the population over 65 years of age. Many different insults appear to be involved in the etiology of the disease, among them environmental toxins and mitochondrial dysfunction. During the past five years, mutations in five different genes have been linked to rare, familial forms of Parkinson's disease. One of the mutated proteins, alpha-synuclein is normally implicated in synaptic plasticity and vesicle function. Dysfunction of this protein might lead to increased cytoplasmic dopamine levels. Since cytoplasmic dopamine is readily prone to autooxidation and enzymatic degradation--processes which generate reactive oxygen species--failure to properly store dopamine into vesicles might lead to oxidative stress. Indeed, nigral tissue from idiopathic Parkinson's disease patients shows signs of oxidative damage. In this article we propose that dopamine-induced oxidative stress might be a common final pathway in the pathogenesis of the disease. PMID- 12739403 TI - [Computerized decision support in drug prescribing I: Better survey of patients' medications yields better quality of care]. AB - New drugs and new discoveries--such as those concerning the use of individual genetic profiles to help customize dosages--have refined drug treatment. Drug companies' marketing efforts as well as patients' views and wishes influence physicians in their choice of treatment. Drug-drug interactions continue to be a common cause of admissions to hospitals. Instantaneous access to a patient's complete drug list and to reliable medical information in a computer-based drug prescribing system will help ensure patient safety. It will also contribute to evidence-based choice of pharmacological principle, drug, method of administration and dosage and to better agreement between patient and doctor. PMID- 12739404 TI - [Computerized decision support in drug prescribing II: A national database to provide up-to-date and unbiased information]. AB - Prescribers today encounter increasing demands for up-to-date knowledge of medical advances and drug therapies, and for a straightforward dialogue with patients. Cost-effective drug treatment calls for fast and intuitive access to information about drugs, treatment strategies and patient data. There are several computer-based drug-prescribing systems in Sweden. Information independent of the drug industry is wanting, as are uniform national standards for medical content and functionality. Decision-support systems must facilitate decisions about therapy, drug distribution and intake, as well as quality work, i.e. support the entire process which defines an effective drug therapy. A very important feature is access to a patient's drug list showing all current drugs. This joint initiative by county councils aims at drawing up a national Swedish specification of requirements for a suitable decision-support system and at creating a national entity responsible for distributing unbiased information from a unique database to a range of computerized medical records systems. PMID- 12739405 TI - [A Cochrane report compares inhaled steroids for asthma. No big differences between the most common preparations]. PMID- 12739406 TI - [Home visits can give family support to those who have recently become parents. First meeting with child health services is important for further contacts]. PMID- 12739407 TI - [Physician, bird photographer and writer Paul Rosenius. His book saved son-in law's life in Goring's Germany]. PMID- 12739408 TI - [An ethically credible intensive care shields the terminally ill patient]. PMID- 12739409 TI - [Reclaim the clinical research--proposal to an action plan]. PMID- 12739410 TI - [Considerations by the local specialty associations should be crucial in the specialty survey]. PMID- 12739411 TI - [The health services can be governed better--if the actors get all the necessary requirements]. PMID- 12739412 TI - [Subutex is the first link in the care continuity of opiate dependence]. PMID- 12739413 TI - [Did the eradication of smallpox open the door for HIV?]. PMID- 12739414 TI - Reflexivity: a personal journey during data collection. AB - In this article Mary Carolan explores the notion of reflexivity as it applies to qualitative research. A study into the experiences of older first-time mothers enabled an exploration of personal views and philosophy in relation to the data collected. PMID- 12739415 TI - The mentor's tale: a reflexive account of semi-structured interviews. AB - In this article, Helen Hand provides an account of personal experience as a novice researcher collecting data using semi-structured interviews. The data were collected as part of a study involving nurse mentors who had had the experience of mentoring and subsequently failing a student nurse. The paper emphasises the importance of reflexivity as a method of signposting to the reader 'what is going on' in the research process. In conclusion, it is suggested that creating the 'joint account' is a complex but worthwhile task, but that great skill is required by the researcher in order to participate in the construction of reality. PMID- 12739416 TI - Ethnography and reflexivity: emotions and feelings in fieldwork. PMID- 12739417 TI - Requesting access for research purposes: issues and guidelines. AB - One of the main considerations in the early stages of the research process is the request for access to research participants. This important stage can yield profitable results or be a source of frustration. Leslie Woods and Paula Roberts discuss issues involved in requesting access, together with suggested guidelines to facilitate the process. PMID- 12739418 TI - Illuminative case study design: a new approach to the evaluation of continuing professional education. AB - Drawing on illuminative evaluation methods originally developed for use in mainstream education, Lorraine Ellis describes a new approach to the evaluation of continuing professional education in nursing through an illuminative case study design. PMID- 12739419 TI - Using conjoint interviews to research the lived experience of elderly rural couples. AB - Understanding the experiences of elderly rural couples in accessing health services can assist nurses and other health professionals in their roles as advocates, service providers, educators, programme planners, and policy makers. In this paper, Frances Racher explores phenomenology as methodology, and its fit with the unstructured conjoint interview as method, in planning for a study to seek knowledge of elderly rural couples' experiences in accessing health services. It is argued that phenomenology is an appropriate approach for gaining an understanding of the experiences of elderly rural couples. In addition, conjoint interviews provide opportunity for the partners to negotiate and jointly construct their responses, when the couple is the unit of study and the couple experience is the topic of interest. As health professionals strive to situate themselves in the everyday worlds of their clients, they are integral in the development and application of new research methods such as the conjoint interview. PMID- 12739420 TI - How to choose and use a computer. PMID- 12739421 TI - Multiorgan autoimmune diseases: a single root for different leaves? PMID- 12739422 TI - [Diabetes and liver diseases]. PMID- 12739423 TI - [Impact of population aging on the social and the health care system: need for a new model of long-term care]. AB - Population aging is characterized by a marked increase in the number of subjects aged 80 years or more (the oldest old). In this group frailty is extremely common. Frailty is a recently identified condition resulting from a severely impaired homeostatic reserve, that places the elderly at the highest risk for adverse health outcomes, including dependency, institutionalization and death, following even trivial events. Geriatric medicine proposes an original methodology for the management of frail elderly subjects, the so called "comprehensive geriatric assessment", as well as a model of long-term care. These have been shown to reduce the risk of hospitalization and nursing home admission, with a parallel decrease in expenses and an improvement in the patient's quality of life. The effectiveness of the long-term care system depends on: 1) the availability of all the services that are necessary for the frail elderly, both in the hospital and in the community; 2) the presence of a coordinating team, the comprehensive geriatric assessment team, that develops and implements the individualized treatment plans, identifies the most appropriate setting for each patient and verifies the outcomes of the interventions; 3) the use of common comprehensive geriatric assessment instruments in all the settings; 4) the gerontological and geriatric education and training of all the health care and social professionals. PMID- 12739424 TI - [Urticaria and angioedema]. AB - Urticaria is a common complaint characterized by the eruption of cutaneous wheals, accompanied by redness and itching and in which mast cells are thought to play a central role. Wheals range from a few millimeters to several centimeters in diameter and are usually short lasting, except in case of urticarial vasculitis in which they last longer than 24 hours. Urticaria may occur alone or be associated with angioedema, that can be defined as a deep dermal and subcutaneous edema typically affecting the lips, face, hands, feet, penis or scrotum. Angioedema may also involve the submucosal tissue, and when the swellings occur in the oropharynx they can be alarming and occasionally life threatening. In many cases the cause of acute urticaria or angioedema may be determined, whilst the pathogenetic and diagnostic aspects of the chronic forms are more complex and variable. The clinical features of, as well as the diagnostic and therapeutic approaches to the different forms of urticaria and angioedema are here reviewed, focusing on chronic idiopathic urticaria. PMID- 12739425 TI - [Oxygen therapy. The new objectives: from survival to quality of life in correlation with state of health]. AB - The aim of this review is to give new insights on the effects, and consequently, on the prescription criteria of long-term oxygen therapy in chronic obstructive pulmonary disease (COPD), on the basis of the evidence accumulated in the recent past. Since the demonstration of the beneficial effect on survival offered by long-term oxygen therapy in severely hypoxemic COPD patients, further potential endpoints of this treatment in such subjects have been progressively evaluated. At present, knowledge of the favorable biochemical effects, especially on the skeletal muscles and brain tissue, and the widespread appreciation of the positive impact on the health-related quality of life parameters provided by long term oxygen therapy in COPD patients should prompt us to reassess its indications in these subjects. PMID- 12739426 TI - [Prevalence and significance of type-2 diabetes mellitus in chronic liver disease, correlated with hepatitis C virus]. AB - A high prevalence of type 2 diabetes mellitus in patients with hepatitis C virus (HCV)-related chronic liver diseases has been reported in numerous studies. Other studies failed to confirm this observation. We have studied the relative prevalence of type 2 diabetes mellitus in two groups of patients respectively presenting with HCV-related chronic liver disease (224 patients) and chronic liver diseases of other etiologies (30 hepatitis B virus-HBV-related chronic liver disease, 22 alcoholic liver cirrhosis), in order to confront them. Our study revealed a higher prevalence of diabetes mellitus in the group of patients with HCV-related chronic liver disease in comparison with the group of patients with chronic liver disease of other etiologies (32.5 vs 15.3%; p = 0.03). Patients with HBV-related liver disease had diabetes in 6.6% of cases, and the difference with patients with HCV-related disease was significant (p = 0.007). Our study confirms a higher prevalence of type 2 diabetes mellitus in patients with HCV-related chronic liver disease. It could be suggested that type 2 diabetes mellitus in patients with HCV-related chronic liver disease could be facilitated by hepatic iron overload and mitochondrial damage. PMID- 12739427 TI - [Osteoarticular scintigraphy in comparison with clinical evidence]. AB - Bone scintigraphy is a technique which is often resorted to in diagnostic rheumatology. There are few data on the effective relevance of bone scintigraphy in the evaluation of chronic inflammatory diseases of the joints. The aim of this study was to compare the results of bone scintigraphy with clinical evidence in patients with rheumatoid arthritis or osteoarthritis. Seventy-five patients were submitted to total body bone scintigraphy (44 rheumatoid arthritis, 31 osteoarthritis). The nuclear medicine specialist indicated the list of joints showing uptake. For the same patients a rheumatologist indicated the number of affected joints. The laboratory and clinical data were recorded. The patients were first stratified according to the prevalence of the clinical evidence and scintigraphic uptake. The distribution was found to be not significant. Only 5.3% of patients showed no uptake. Thirty-three patients had no clinical evidence of disease; among these, 30 showed joint uptake. Considering only the patients with clinical evidence, 97.6% showed joint uptake. These results were confirmed even when the data were analyzed by sex, disease and therapy. Considering the patients with clinical evidence, the uptake/clinical ratio did not show any significant correlation. The number of joints with clinical evidence correlated with the erythrocyte sedimentation rate. The number of joints showing uptake correlated only with age. In conclusion, on average, scintigraphy, performed in patients with rheumatoid arthritis and osteoarthritis, highlights a significantly higher number of joints involved as compared to what would be expected on the basis of clinical evaluation. It remains to be defined whether this is an overestimation related to the characteristics of the scan or whether it is sign of a higher sensitivity in highlighting the site of inflammation. Against the latter hypothesis is the absence of correlation with the inflammatory indexes. PMID- 12739428 TI - Heart rate variability in cluster headache. AB - Eight patients with cluster headache were studied by evaluating the heart rate variability on the basis of 24-hour ECG monitoring performed during attacks (critic periods) to demonstrate if any imbalance of the autonomic nervous system was present and to determine its temporal correlation, if any, with the attack. The same monitoring allowed us to compare the patients' data during headache-free periods (intercritic periods) with those of normal controls. Our investigation demonstrated a severe sympathovagal imbalance during spontaneous attacks: the parasympathetic drive is at once increased at the onset with a mild reduction of the sympathetic drive, and all these modifications slowly disappear at the end of each attack. Comparison of the average low-frequency and high-frequency values during intercritic periods showed a significant reduction in the low-frequency values in patients, even when the low-frequency/high-frequency ratio was normal. This is highly suggestive of the presence of an autonomic nervous system dysfunction. PMID- 12739429 TI - [Syndrome of cholesterol crystal embolism: clinical case]. AB - Cholesterol crystal embolization syndrome is a multiorgan disease that frequently occurs as a complication of invasive cardiovascular procedures and of thrombolytic or anticoagulant therapy. The symptoms are due to the displacement of atheromatous material dislodged from unstable or injured, by mechanical manipulation, plaque to arteriolar vessels. The real incidence of cholesterol embolization is not known. Often the diagnosis is missed because of the time between intervention and clinical findings and because the organs involved can be many and various. The most common clinical manifestations are acute renal failure and hypereosinophilia. The prognosis is poor and the mortality high because of the progression of renal failure. In this case report we present the clinical history of a 62-year-old male patient with a history of cigarette smoking and hypertension who was submitted to emergency surgery following the acute dissection of a type A aortic aneurysm. About 2 weeks after surgical intervention the patient developed a multiorgan disorder (gastroenteric, neuromuscular and renal involvement) associated with hypereosinophilia and with increased levels of the markers of inflammation. The symptoms were transient and probably due to embolization of cholesterol crystals; no specific therapy was administered. On the other hand, no therapeutic regimen has been codified to date. The best clinical approach is prophylactic, that means identifying those patients who are at high risk for an invasive vascular procedure. PMID- 12739431 TI - Exercise and your health. How much is enough? PMID- 12739432 TI - Health tips. Nondairy calcium. PMID- 12739430 TI - [Multiorgan autoimmune syndrome: case report]. AB - The present case report refers to a multiorgan autoimmune disease manifesting following thymectomy performed for a benign thymoma. This disease is characterized by hypothyroidism, severe myasthenia, polymyositis and alopecia which are organ-specific diseases probably with a different time of onset but which are all an expression of the same immunopathologic process occurring in individuals who have a genetic predisposition. Characteristic of the present case is not only the association of the different immunopathologic clinical pictures but also the rather difficult differential diagnosis between a hypothyroidism related myopathy and polymyositis. It was possible to formulate the diagnosis by integrating the results of clinical and laboratory evaluation with the therapeutic outcome. The onset of the syndrome was attributed to the withdrawal, following surgery, of the inhibitory effects of the thymoma on some clones of autoreactive lymphocytes. PMID- 12739433 TI - Opioid medicines for chronic low back pain. PMID- 12739434 TI - New hand-hygiene guidelines issued. PMID- 12739436 TI - Hallucinations. May signal larger problem. PMID- 12739435 TI - Hemorrhoids and anal tears. Sometimes painful. PMID- 12739437 TI - Thyroid testing. Chemical messengers. PMID- 12739438 TI - Second opinion. I know the benefits of eating fish that's high in omega-3 fatty acids. Yet, in restaurants, the fish on the menu is often deep-fried or baked with butter. What are healthy ways to order fish in restaurants? PMID- 12739439 TI - Second opinion. What tests do blood banks do on donated blood? PMID- 12739440 TI - The heart's back-up system saves lives. And a search for ways to make new cardiac vessels sprout. PMID- 12739441 TI - Wakeful patients aid surgeons during a procedure to prevent stroke. PMID- 12739442 TI - Another test, another disappointment for chelation therapy. PMID- 12739443 TI - Protecting rheumatoid arthritis sufferers from heart attacks. PMID- 12739445 TI - CVD mortality rates decline. Life expectancy rises as a result; typical patients are older. PMID- 12739444 TI - More than two kids may mean more risk for mom--and dad. PMID- 12739446 TI - Solving the puzzle of diabetes. New strategies emerge to help fight a threat to your heart. PMID- 12739447 TI - How to keep recovery on track. PHone reminders and frank talk help lock in health gains. PMID- 12739448 TI - Ask the doctors. My wife is a relatively healthy middle-aged woman, so I was surprised when she recently had a heart attack. Isn't that unusual for a woman? PMID- 12739449 TI - Ask the doctors. I have arthritis and take Motrin. Is there a problem with also taking aspirin, since my doctor has told me to do this ever since my heart attack? PMID- 12739450 TI - Ask the doctors. I am 80 years old and have always been healthy, other than some high blood pressure. During a routine check-up, my family doctor discovered that my cholesterol level was elevated, and started me on medication. What is the point of taking cholesterol medications at my age? PMID- 12739451 TI - Aspirin curbs colon polyps. But for many, bleeding risks may offset benefits. PMID- 12739453 TI - More hope for heart failure. Findings suggest expanded use of aldosterone blockers. PMID- 12739452 TI - Sugar's not-so-sweet revenge. Avoid added sugar, say nutrition experts. PMID- 12739454 TI - Hormones and mood. PMID- 12739455 TI - Exercise helps before, after hip replacement. PMID- 12739456 TI - Prolonged warfarin thwarts return of clots. PMID- 12739457 TI - How contagious is it? PMID- 12739458 TI - Megadose vitamin D foils fractures? PMID- 12739459 TI - Topical glucosamine for osteoarthritis? PMID- 12739460 TI - NSAIDs and your heart: risky or not? PMID- 12739461 TI - Opioids ease neuropathic pain, but... PMID- 12739462 TI - Medication mix-ups. PMID- 12739463 TI - DES may be forgotten, but it's not gone. PMID- 12739464 TI - I'm 56 years old and have total cholesterol of 217 milligrams per deciliter, LDL of 131 mg/dl, HDL of 67 mg/dl, and a total-to-HDL ratio of 3.24. My doctor says that ratio is good for a woman my age, so why does she want me to lower my LDL? PMID- 12739466 TI - Hope for children with peanut allergy. PMID- 12739465 TI - Is it better to cover a cut or scrape with a bandage or let it heal uncovered? And are those new waterproof or liquid bandages any better than the old ones? PMID- 12739467 TI - Changing sleep patterns in infants and children. PMID- 12739468 TI - What to consider when a child has low back pain. PMID- 12739469 TI - When one sibling is injured, another one is at risk--for a while. PMID- 12739470 TI - Can parents read instructions for installing child safety seats? PMID- 12739471 TI - Helping children through stressful times...and it's not too late for TV turnoff. PMID- 12739472 TI - Product recalls. Recalls: National animal swing roundup... Graco SnugRide car seats...and girls' hooded winter jackets. PMID- 12739473 TI - Treatment interruption; advanced HIV; new ideas: Dr. Cal Cohen interview on retrovirus conference (part 2 or 2). Interview by John S. James. AB - The conclusion of Dr. Cohen's interview includes an in-depth look at current thinking on treatment interruption, strategies for highly treatment-experienced patients, and other topics from the recent Retroviruses conference. PMID- 12739474 TI - SARS Web information. AB - Where to find current information about Severe Acute Respiratory Syndrome (SARS), a new disease that can cause severe or fatal pneumonia. PMID- 12739475 TI - Abacavir arm stopped in clinical trial. AB - A government clinical trial stopped treatment with abacavir plus AZT plus 3TC, due to more viral rebound than other treatments in the study. Many unknowns remain. PMID- 12739476 TI - Huge Medicaid cuts weighed in Washington. AB - A $92 billion Medicaid cut passed by the U.S. House of Representatives, and a plan by the Bush Administration to reduce entitlement funding of medical care for the poor and elderly and give states more power to choose whose benefits are ended, could seriously reduce HIV care in the coming decade. PMID- 12739477 TI - Africa activism: money for AIDS not for war; South Africa antiretrovirals. AB - Africa Action seeks organizational endorsements for prioritizing an epidemic that has already killed 25 million people. Separately, South Africa's Treatment Action Campaign seeks international support for government cooperation in making antiretroviral treatment available there. PMID- 12739478 TI - Coping with loneliness and HIV. PMID- 12739479 TI - Social interventions, community, and HIV prevention. PMID- 12739481 TI - Physician burnout and the social contract. PMID- 12739480 TI - Easing HIV provider burnout: an in vivo approach. PMID- 12739482 TI - HHS awards $1 billion in Ryan White grants. PMID- 12739483 TI - Politics. New American 'science' worries global researchers, activists. PMID- 12739484 TI - Disclosure rules. Advocates: HIPAA brings end to U.S. medical privacy. PMID- 12739485 TI - Groups sues to block HIPAA privacy regulations. PMID- 12739486 TI - Statute of limitations invalidates viatical agreement claim. PMID- 12739487 TI - Immigrant denied deportation relief despite HIV medical need. PMID- 12739488 TI - Emory law school conference tackles access to generic AIDS drugs. PMID- 12739489 TI - Success of needle exchanges varies within the program. PMID- 12739490 TI - Criminal exposure. Felony conviction of HIV transmission upheld. PMID- 12739491 TI - Criminal transmission. States crack down on nondisclosing partners. PMID- 12739492 TI - 10 years added to sentence for criminal exposure to HIV. PMID- 12739493 TI - Criminal transmission. HIV-positive prostitute sentenced to 10 years. PMID- 12739495 TI - As medical marijuana battle heats up, doctors take stand. PMID- 12739494 TI - GOP works to halt medical marijuana movement. PMID- 12739496 TI - Treatment legislation. Bipartisan bill for early HIV treatment introduced in Senate. PMID- 12739497 TI - Emerging drugs in the fight against HIV disease. PMID- 12739498 TI - Report from Boston: the 10th Conference on Retroviruses and Opportunistic Infections (CROI). PMID- 12739499 TI - Treatment of tuberculosis in the HIV-infected patient. PMID- 12739500 TI - Immune-based therapy in HIV. PMID- 12739501 TI - When to start HAART, and what to start. PMID- 12739502 TI - Taking HAART to heart: antiretroviral toxicities. PMID- 12739503 TI - Drug resistance and treatment of experienced patients. PMID- 12739504 TI - HIV and hepatitis. PMID- 12739505 TI - New York City budget crisis. Will disorganization and higher property taxes push people with HIV into the streets? PMID- 12739506 TI - Meeting clients "where they are". BP's David Pratt speaks with Don McVinney of the Harm Reduction Coalition. PMID- 12739507 TI - As easy as ESAP. The New York State Expanded Syringe Access Demonstration Program. PMID- 12739508 TI - My journey of transformation. Reflections on vision impairment caused by HIV. PMID- 12739509 TI - "When it hurts to ask for help". PMID- 12739510 TI - Sex, research & community. PMID- 12739512 TI - [Is noncompliance with the work schedule regulation cause for criminal apprehension?]. PMID- 12739511 TI - [Experimental examination concerning the efficacy of silver-coated Dacron prostheses in vascular graft infections following subcutaneous implantation in a standardized infection model]. AB - It was the aim of the study to examine the efficacy of silver coated prostheses in comparison to Rifampin in impregnated prostheses in the prevention of vascular graft infections. MATERIAL AND METHODS: 24 C3H/HcN mice with a bodyweight between 24 and 27 grams were assigned to four different groups. GROUP I: control gel sealed Dacron graft (Uni-Graft DV) (6), GROUP II: gel-sealed Dacron graft (Uni Graft DV) contaminated locally with 2 x 10(7) CFU/1.2 ml Staphylococcus aureus ATCC 25923 (6), GROUP III: silver prosthesis (Intergard Silver) contaminated locally with 2 x 10(7) CFU/0.2 ml Staphylococcus aureus ATCC 25923 (6), GROUP IV: Rifampin impregnated prosthesis contaminated locally with 2 x 10(7) CFU/0.2 ml Staphylococcus aureus ATCC 25923 (6). 14 days after primary operation all animals were euthanized and the grafts harvested. Specimens were examined for signs of infections by histology and microbiology. RESULTS: At termination of the trial on day 14 none of the grafts of group I were contaminated. 6 out of 6 grafts in group II, 6 out of 6 grafts in group III and 1 out of 6 grafts in group IV presented with infected grafts. The use of antimicrobial Rifampin could significantly prevent infection after bacterial challenge in group IV. CONCLUSION: The silver protected prosthesis (Intergard Silver) seems to be not effective in protecting vascular infection in vivo. However, the Rifampin group showed excellent results. In conclusion Rifampin bonded gelatin-sealed Dacron grafts are significantly more resistant to bacteremic infection than are silver/collagen-coated Dacron grafts. PMID- 12739513 TI - [Summary of the coalition agreement on health policy]. PMID- 12739514 TI - [Generally effective for unstable bladder? Tamsulosin: application not limited to benign prostatic hyperplasia]. PMID- 12739515 TI - [Renal surgery--triumph for laparoscopy?]. PMID- 12739516 TI - [Toxicologic Information and Documentation Society Research Project introduces results. New information system of chemical products and analysis for practical application]. PMID- 12739517 TI - [Surgery of the upper gastrointestinal tract (part 1)]. PMID- 12739518 TI - [Report on the 90th meeting of the "Vereinigung Mittelrheinischer Chirurgen vom 19.-21. September 2002 in Bruschal"]. PMID- 12739520 TI - Should rationing of health care be explicit? AB - The aim of this paper is to discuss whether medical professionals should inform patients that there may be drugs or procedures that would benefit them., but which the National Health Service cannot afford to provide. The main body of the paper examines the argument put forward by Professor David Hunter who suggests that rationing of health care should be implicit. I will argue that his arguments are unconvincing. Some of the reasons that Coast and Mechanic give support to implicit rationing of health care are looked at briefly and similarly found to be wanting. In our present culture openness is encouraged from doctors. It is also acknowledged by the medical profession that patients' autonomy should be respected. The importance of autonomy is emphasised in the teaching of our medical students. My submission will be that it is difficult to see how patients' autonomy can properly be exercised unless they are told that treatment is being refused to them that might help their condition. PMID- 12739519 TI - General practitioners and predictive genetic testing for late-onset diseases in Flanders: what are their opinions and do they want to be involved? AB - OBJECTIVE: Investigate the attitudes of general practitioners (GPs) concerning predictive testing for late-onset diseases, as well as the perception of their own role in this context. METHODS: 356 GPs received mail questionnaires with telephone pre-notifications and reminders. RESULTS: The questionnaire was returned by 60% (n=215). The GPs' attitudes toward predictive testing for breast cancer, thyroid cancer, Alzheimer disease and Huntington's disease were influenced by the availability and the type of preventive and therapeutic options, the age of onset of the disease as well as by ethical concerns. Regarding a possible tasks for GPs, most of the GPs focussed on gate-keeping aspects, such as providing information and making referrals. CONCLUSION: The GPs were supportive of a limited role for general practice in predictive testing. Genetic education for GPs is needed, with attention to non-directiveness and the characteristic psychosocial and ethical implications of this particular type of genetic testing. PMID- 12739521 TI - Control of patient information. PMID- 12739522 TI - [Effect of national and local guidelines on medical practice. Experience with 30 years of internal guidelines in hematology]. PMID- 12739523 TI - [77th annual meeting of the Japanese Association for Infectious Diseases. Fukuoka, Japan. April 17-18, 2003. Abstracts]. PMID- 12739524 TI - Clinical ethics committees in the UK. PMID- 12739526 TI - Current readings in nuclear medicine. PMID- 12739525 TI - [Open letter to German women physicians and men physicians]. PMID- 12739527 TI - Stroke, cognitive decline and vascular dementia: the silent epidemic of the 21st century. PMID- 12739529 TI - [The 103rd annual meeting of the Japan Surgical Society. Hokkaido, Japan. June 4 6, 2003. Programs]. PMID- 12739528 TI - [Early diagnosis: is information a value itself? Evaluation of genetic screening]. PMID- 12739530 TI - [Consensus on the bases of cooperation of gastroenterologists and visceral surgeons]. PMID- 12739531 TI - [The 107th annual meeting of the Japanese Ophthalmological Society. Fukuoka, Japan. April 17-20, 2003. Abstracts]. PMID- 12739532 TI - Diane Pretty: final acts. PMID- 12739533 TI - Jesse's intent. PMID- 12739534 TI - Integrating complementary and alternative medicine: fresh challenges for RECs. PMID- 12739535 TI - No public health without public death. PMID- 12739536 TI - Multifactorial intervention and cardiovascular disease in type 2 diabetes. PMID- 12739537 TI - Assisted reproduction in Italy. PMID- 12739539 TI - Clinical trials: the intersection between research and medicine. PMID- 12739540 TI - Human subject protections in the United States: perspectives from the Office for Protection from Research Risks. AB - The U.S. Department of Health and Human Services (DHHS) regulations require that all human subjects research supported by DHHS be reviewed and approved by a local institutional review board (IRB). With few exceptions, investigators may not involve human subjects in research without their informed consent, and additional safeguards are required when subjects are likely to be vulnerable to coercion or undue influence. Institutions that receive DHHS funding must enter into an "Assurance" of compliance with the Office for Protection from Research Risks (OPPR), which has the authority for oversight and implementation of the human subjects regulations. As discussed more fully below, Assurances are of a contract nature in that they formally commit the institutions to adherence to the regulations and the ethics standards relevant to research on human subjects. This article addresses the application of human subject protections in biomedical research. PMID- 12739541 TI - Modeling of genetic instability--looking for "Newton's binomial" of colon cancer. PMID- 12739542 TI - United States regulatory requirements for research involving human subjects. PMID- 12739543 TI - The National Bioethics Advisory Commission: bridging the tension between scientific and public policy analysis. AB - During the period between the early 1980s to the mid 1990s, the U.S. was distinguished from most other developed countries by its lack of a national-level public body to assist the government in its policy-making on topics of biomedical ethics. While Canada, Denmark, France, Spain, and other countries regularly sought advice from public commissions on issues ranging from reproductive technologies to euthanasia, the U.S. relied on myriad state commissions, court decisions, and academic bodies. The result was a pattern of policy-making that was slower and more unpredictable than that of its peers. With the 1996 appointment of the National Bioethics Advisory Commission (NBAC) by President Clinton, there has been a change in the process of U.S. public policy development. This article provides an overview of the NBAC and highlights recent areas of focus and related recommendations. PMID- 12739545 TI - Biotechnology and health. PMID- 12739544 TI - The ethics of biotechnology. PMID- 12739546 TI - No privacy for all? Serious failings in the HHS medical records regulations. AB - The Bush administration surprised many by the approval without major revisions of the Clinton administration's HHS medical records regulations, despite heavy lobbying from the health industry. Though these "privacy rules" are supposed to protect patient confidentiality, what has gone unmentioned are the regulations' major lapses that breach informed consent and confidentiality. Recently issued "clarifications" of the regulations reveal that they do not prevent unconsented access to sensitive medical information by marketers, health plans, health care clearinghouses, and law enforcement. These problems with the regulations constitute a serious breach of patient privacy, endangering the doctor-patient relationship and potentially driving up health care costs, and need to be addressed. PMID- 12739547 TI - EuropaBio's ethics committee and Advisory Group on Ethics(AGE): an overview. PMID- 12739548 TI - Ethical, social and public awareness issues in gene therapy EuropaBio. AB - EuropaBio, the European Association for Bio-industries, represents 40 corporate members operating world wide and 14 national associations (totaling up to 700 small- and medium-sized enterprises) involved in research, development, testing, manufacturing, sales, and distribution of biotechnology-derived products and services in the field of health cae, agriculture, food, and the environment. AGE is a group of researchers and university professors involved in high-level professional activities related to bioethics, and particularly interested in ethical issues related to the development and use of modern technology. It is essential that industry actively participates in, and contributes to, the social debate on emerging technologies. Therefore, EuropaBio presents herein its view on gene therapy and its responsible development and use. PMID- 12739549 TI - Initial phase III AIDSVAX results do not reach primary end point. PMID- 12739550 TI - Placement of self-expanding metallic stents in the pancreatic duct for treatment of obstructive complications of pancreatic cancer. AB - BACKGROUND: Stents have been placed through malignant pancreatic strictures, mainly to alleviate pain of presumed obstructive origin. Self-expanding metallic stents have major advantages over plastic stents when used for treatment of malignant biliary strictures. However, there are few reports of their use in patients with malignant pancreatic duct strictures, especially those with complications related to ductal obstruction. METHODS: Self-expanding metallic stents were placed in the pancreatic ducts of 3 patients with obstructive complications of pancreatic cancer: smoldering pancreatitis, a disrupted pancreatic duct with pseudocyst caused by open surgical biopsy, and a disrupted pancreatic duct with fistula and resultant liver abscess. All 3 patients had metallic stents placed concomitantly in the biliary tract; one had enteral stents placed as well. Clinical and pathology records and imaging studies were reviewed retrospectively. OBSERVATIONS: In all cases, there was resolution of the specific clinical problem and reasonable survival (1.5 years in one patient). CONCLUSIONS: The use of self-expanding metallic stents for treatment of certain obstructive complications of pancreatic tumors is feasible and effective. PMID- 12739551 TI - FDA aims to improve and streamline the drug development and review process. PMID- 12739552 TI - Endoscopic features of esophageal tuberculosis. AB - BACKGROUND: Esophageal tuberculosis is rare. Clinical and endoscopic features are variable, diverse, nonspecific, and poorly described. These findings may be confused with those of esophageal cancer and deep fungal infection. METHODS: Medical records from a 5-year period (January 1997 to December 2001) were searched for cases of esophageal tuberculosis. For identified cases, the clinical, radiologic, and endoscopic features were evaluated. OBSERVATIONS: Four cases of esophageal tuberculosis were encountered during the 5-year period studied. Three of the patients presented with dysphagia and weight loss. One patient had aspiration pneumonia and another fatal hematemesis. Endoscopic features included deep and large proximal esophageal ulcers in 2 patients, tracheo-esophageal fistula in one, and nonhealing proximal esophageal ulcer in another patient. CONCLUSION: The present case series indicates that certain endoscopic features, such as deep and large esophageal ulcers, tracheoesophageal fistula, and nonhealing ulcer, are strongly suggestive of tuberculosis-related esophageal lesions. PMID- 12739553 TI - Clearance of senescent erythrocytes: wheat germ agglutinin distribution on young and old human erythrocytes. AB - To add an additional aspect to the process of recognition and removal of senescent human erythrocytes from the circulation, the binding of wheat germ agglutinin (WGA) to separated young, old and sialidase-treated human erythrocytes is evaluated with the immune-electron microscopical method. WGA/gold conjugate binding to old erythrocytes was lower (27%) than to young erythrocytes and even lower following treatment with sialidase (82%), exhibiting a clustered, non continuous labeling pattern in all three erythrocyte populations, thus showing a possible redistribution of WGA binding sites. The decrease in bound WGA/gold particles correlates well with the previously reported decrease in surface sialic acid on old erythrocytes. The binding of WGA/gold are indicative of the changes occurring on erythrocyte membrane surfaces when interacting with different agglutinins. PMID- 12739554 TI - Intrahepatic placement of a PEG tube. PMID- 12739555 TI - NME digest. PMID- 12739556 TI - Successful endoscopic treatment of a complete transection of the bile duct complicating laparoscopic cholecystectomy. PMID- 12739557 TI - Percutaneous transhepatic choledochoscopic injection of ethanol with OK-432 mixture for palliation of malignant biliary obstruction. PMID- 12739558 TI - Treatment of primary malignant melanoma of the esophagus with endoscopic injection of interferon-beta combined with systemic chemotherapy: a case report. PMID- 12739559 TI - Endoscopic "clip and cut" diverticulotomy for a giant midesophageal diverticulum. PMID- 12739560 TI - Endoscopic management of Kock pouch dysfunction: case report of a method to establish wire-guided pouch access for catheterization. PMID- 12739561 TI - Cecal botryomycosis: case report and review. PMID- 12739562 TI - Endoscopic evidence of GI hemorrhage induced by 5-fluorouracil administration. PMID- 12739563 TI - Endoscopic drainage of a pancreatic pseudocyst in a symptomatic patient with subtotal gastrectomy and Roux-en-Y anastomosis. PMID- 12739564 TI - [Management of the patient with mucoviscidosis]. PMID- 12739565 TI - Retroperitoneal neurilemoma diagnosed by EUS-guided FNA. PMID- 12739566 TI - Line in the sand. PMID- 12739567 TI - Cap polyp of the esophagus caused by cholesterol embolization. PMID- 12739568 TI - Herpetic esophagitis and intractable hiccups (singultus) in an immunocompetent patient. PMID- 12739569 TI - Endoscopic therapy is effective for bleeding ileal ulcer. PMID- 12739570 TI - Endoscopic diagnosis of Waldenstrom's macroglobulinemia masquerading as chronic diarrhea. PMID- 12739571 TI - Pancreatic ascites treated by endoscopic pancreatic sphincterotomy alone: a case report. PMID- 12739572 TI - Highlights of selected US and international groups providing HIV care and services in developing countries. PMID- 12739573 TI - Video cholangioscopy with a new choledochoscope: a case report. PMID- 12739574 TI - Endoscopic cystenterostomy of non-bulging pancreatic fluid collections without EUS: do we really need a communication between the cyst and the duct? PMID- 12739575 TI - Primary prophylaxis against bleeding from gastric fundal varices. PMID- 12739576 TI - Endocrine-related resources from the National Institutes of Health. PMID- 12739578 TI - Dr Peter Nathan, M.D., F.R.C.P. PMID- 12739579 TI - Challenging Food and Drug Administration interpretations of the Federal Food, Drug, and Cosmetic Act. PMID- 12739580 TI - Underdeveloped: FDA's authority to take photographs during an FDA establishment inspection under section 704. PMID- 12739581 TI - CFSAN's program priorities: from food safety to food security. PMID- 12739582 TI - Direct-to-consumer marketing: the Food and Drug Administration is not alone. PMID- 12739583 TI - The legal status and regulatory context of "health foods" in the European Union. PMID- 12739584 TI - The Drug Price Competition and Patent Term Restoration Act of 1984: fine-tuning the balance between the interests of pioneer and generic drug manufacturers. PMID- 12739585 TI - Regulating the fate of pharmaceutical drugs: a new prescription for the environment. PMID- 12739586 TI - Bibliography. Current world literature. Skin and soft tissue infections. PMID- 12739587 TI - Bibliography. Current world literature. Respiratory infections. PMID- 12739588 TI - The future of human cloning: prescient lessons from medical ethics past. PMID- 12739590 TI - Organ transplantation at the millennium: regulatory framework, allocation prerogatives, and political interests. PMID- 12739591 TI - I want a girl (boy) just like the girl (boy) that married dear old dad (mom): cloning lives. PMID- 12739592 TI - How pharmacogenomics will impact the federal regulation of clinical trials and the new drug approval process. PMID- 12739593 TI - A feminist understanding of sex-selective abortion: solely a matter of choice? PMID- 12739594 TI - Pennsylvania takes the lead with new federal health care tax credit program. PMID- 12739596 TI - [Subacute de Quervain thyroiditis]. PMID- 12739597 TI - Report from the Task Force of the European Society of Cardiology for the interpretation of the neonatal electrocardiogram. PMID- 12739598 TI - [Treatment of anal fissure]. PMID- 12739599 TI - The BSAVA presents its awards. PMID- 12739600 TI - International Animal Health, disease surveillance and trade in 2002. PMID- 12739601 TI - Airborne transmission of foot-and-mouth disease virus from Burnside Farm, Heddon on-the-Wall, Northumberland, during the 2001 epidemic in the United Kingdom. AB - The results of a detailed assessment of the atmospheric conditions when foot-and mouth disease (FMD) virus was released from Burnside Farm, Heddon-on-the-Wall, Northumberland at the start of the 2001 epidemic in the UK are consistent with the hypothesis that the disease was spread to seven of the 12 farms in the immediate vicinity of the source by airborne virus, and airborne infection could not be ruled out for three other premises; the remaining two premises were unlikely to have been infected by airborne virus. The distances involved ranged from less than 1 km up to 9 km. One of the farms which was most probably infected by airborne virus from Burnside Farm was Prestwick Hall Farm, which is believed to have been key to the rapid spread of the disease throughout the country. In contrast, the results of detailed atmospheric modelling, based on a combination of clinical evidence from the field and laboratory experiments have shown that by assuming a relationship between the 24-hour average virus concentrations and subsequent infection, threshold infection levels were seldom reached at the farms close to Burnside Farm. However, significant short-term fluctuations in the concentration of virus can occur, and short-lived high concentrations may have increased the probability of infection and explain this discrepancy. PMID- 12739602 TI - Influence of detomidine and buprenorphine on motor-evoked potentials in horses. AB - Horses need to be sedated before they are investigated by transcranial magnetic stimulation because of the mild discomfort induced by the evoked muscle contraction and the noise of stimulation. This paper describes the influence of a combination of detomidine (10 microg/kg bodyweight) and a low dose of buprenorphine (2.4 microg/kg) on the onset latency and peak-to-peak amplitude of magnetic motor-evoked potentials in normal horses. There were no significant differences between measurements of these parameters made before the horses were sedated and measurements made 10 and 30 minutes after the drugs were administered. PMID- 12739603 TI - Analysis of passive surveillance data for antimicrobial resistance from cases of neonatal bovine enteritis. PMID- 12739604 TI - Congenital urethral obstruction, bilateral hydronephrosis and uroperitoneum in a lamb. PMID- 12739605 TI - Vaccine-associated feline sarcoma. PMID- 12739606 TI - Crushed tail head syndrome in cattle. PMID- 12739607 TI - Molecular detection and characterisation of Taylorella equigenitalis. PMID- 12739608 TI - FLUTD and transitional cell carcinoma of the bladder. PMID- 12739609 TI - Fees for retired RCVS members. PMID- 12739610 TI - Rabies experiences in Morocco. PMID- 12739611 TI - Septation and separation within the outflow tract of the developing heart. AB - The developmental anatomy of the ventricular outlets and intrapericardial arterial trunks is a source of considerable confusion. First, major problems exist because of the multiple names and definitions used to describe this region of the heart as it develops. Second, there is no agreement on the boundaries of the described components, nor on the number of ridges or cushions to be found dividing the outflow tract, and the pattern of their fusion. Evidence is also lacking concerning the role of the fused cushions relative to that of the so called aortopulmonary septum in separating the intrapericardial components of the great arterial trunks. In this review, we discuss the existing problems, as we see them, in the context of developmental and postnatal morphology. We concentrate, in particular, on the changes in the nature of the wall of the outflow tract, which is initially myocardial throughout its length. Key features that, thus far, do not seem to have received appropriate attention are the origin, and mode of separation, of the intrapericardial portions of the arterial trunks, and the formation of the walls of the aortic and pulmonary valvar sinuses. Also as yet undetermined is the formation of the free-standing muscular subpulmonary infundibulum, the mechanism of its separation from the aortic valvar sinuses, and its differentiation, if any, from the muscular ventricular outlet septum. PMID- 12739612 TI - Vascular adaptations for heat conservation in the tail of Florida manatees (Trichechus manatus latirostris). AB - Although Florida manatees (Trichechus manatus latirostris) have relatively low basal metabolic rates for aquatic mammals of their size, they maintain normal mammalian core temperatures. We describe vascular structures in the manatee tail that permit countercurrent heat exchange (CCHE) to conserve thermal energy. Approximately 1000 arteries juxtaposed to 2000 veins are found at the cranial end of the caudal vascular bundle (CVB); these numbers decrease caudally, but the 1:2 ratio of arteries to veins persists. Arterial walls are relatively thin when compared to those previously described in vascular countercurrent heat exchangers in cetaceans. It is assumed that CCHE in the CVB helps manatees to maintain core temperatures. Activity in warm water, however, mandates a mechanism that prevents elevated core temperatures. The tail could transfer heat to the environment if arterial blood delivered to the skin were warmer than the surrounding water; unfortunately, CCHE prevents this heat transfer. We describe deep caudal veins that provide a collateral venous return from the tail. This return, which is physically outside the CVB, reduces the venous volume within the bundle and allows arterial expansion and increased arterial supply to the skin, and thus helps prevent elevated core temperatures. PMID- 12739613 TI - The equine hind limb is actively stabilized during standing. AB - Horses spend much of their life standing, and they are believed to be able to keep their limbs straight without muscular effort. We tested the hypothesis that the stifle (knee) and hock (tarsal) joints could be stabilized merely with the help of a passive lock mechanism whereby the patella is secured behind a hook, formed by the medial femoral trochlea. In anaesthetized animals and isolated limbs the stifle and hock flex readily under compression. In isolated limbs this collapse was prevented by a small force applied to the patella, mimicking the action of the vastus medialis muscle. In vivo, when the limb was planted loosely on the ground none of the muscles with a connection to the patella was active. However, during weight-bearing the vastus medialis (but no other muscle) was active, providing the necessary traction to stabilize the stifle. The required tension was estimated to be less than 2% of the force that would be needed in absence of a lock mechanism. Diagnosis and treatment of patellar fixation should include the possibility of overactive vastus medialis muscle as a possible cause of the disorder. PMID- 12739614 TI - Muscle fibre breakdown in venom-induced muscle degeneration. AB - We studied the early stages of the degeneration of skeletal muscles using the venom of Notechis scutatus as the myotoxic agent. The venom was used at a dose equivalent to the LD50 in the mouse. There was no mortality amongst the rats. Electron microscopy was used to show the progressive hypercontraction of sarcomeres and the loss of alignment of myofibrils in individual muscle fibres. Between areas of hypercontraction sarcomeres were torn, shedding loosened myofilaments into the cytosol. Western blotting and Coomassie staining were used to compare the respective rates of loss of desmin, titin, actin, myosin and dystrophin. We showed that desmin and titin were the first proteins to be degraded with a time to 50% loss of approximately 1 h and 3 h, respectively. The loss of major contractile proteins, myosin and actin, was rather slower. The loss of dystrophin was also slower than the loss of desmin and titin. Early damage to the plasma membrane of the muscle fibre caused the cells to depolarize, probably promoting the hypercontraction of the sarcomeres, but actual loss of membrane was incomplete even at 24 h. We suggest that the early degradation of desmin and titin was responsible for the disaggregation of the sarcomeres; the liberated contractile proteins myosin and actin were shed into the cytosol, where they were degraded. Phagocytic cells that had invaded the degenerating muscle fibres were primarily involved in the clearance of damaged mitochondria. PMID- 12739615 TI - Immunocytochemical characteristics of elbow, knee and ankle muscles of the five toed jerboa (Allactaga elater). AB - Biochemical adaptations of limb myofibres to intensive bipedal hopping were investigated using the five-toed jerboa Allactaga elater as a model in comparison with the rat. Immunofluorescence methods included immunoreactivity to anti-fast and anti-slow MHC and troponin I. There is no specialization of triceps caput mediale for postural function in the minute non-locomotor forelimbs, unlike quadruped mammals. The various elbow extensor heads and the flexor muscles are alike with regard to fibre type population and cross-sectional areas of each type of fibre. The extensor muscle in the elongated hindlimbs of the five-toed jerboa, at both the knee and the ankle joints, differ from each other extensively. One head, made up of an extremely high percentage of type I, fatigue-resistant fibres, is suited to postural function. Two extensor heads at each joint contain a very high percentage of type IIB fibres (having the greatest maximal velocity of contraction) and are able to produce the powerful acceleration needed to trigger the leap. The relative cross-sectional areas of the myofibres are characteristic of hopping locomotion: predominance in number of one type of myofibre in a muscle accompanies greater cross-sectional area, which increases muscle efficiency in either postural or accelerative function of the muscle. PMID- 12739616 TI - Early GABA(A) receptor clustering during the development of the rostral nucleus of the solitary tract. AB - While there is an abundance of gamma-aminobutyric acid (GABA) in the gustatory zone of the nucleus of the solitary tract of the perinatal rat, we know that GABAergic synapse formation is not complete until well after birth. Our recent results have shown that GABA(B) receptors are present at birth in the cells of the nucleus; however, they do not redistribute and cluster at synaptic sites until after PND10. The present study examined the time course of appearance and redistribution of GABA(A) receptors in the nucleus. GABA(A) receptors were also present at birth. However, in comparison to GABA(B) receptors, GABA(A) receptors underwent an earlier translocation to synaptic sites. Extrasynaptic label, for example, of GABA(A) receptors was non-existent compared to GABA(B) receptors at PND10 and well-defined clusters of GABA(A) receptors could be seen as early as PND1. We propose that while GABA(A), receptors may play an early neurotransmitter role at the synapse, GABA(B) receptors may play a non-transmitter neurotrophic role. PMID- 12739617 TI - Pollical oblique ligament in humans and non-human primates. AB - A morphological study of the oblique ligament in the thumb is presented. The ligament was consistently described in human specimens and compared with dissections of non-human primates from different species. The oblique ligament was found in some, but not all, specimens in each of the following species examined: chimpanzee, orangutan, gibbon, anubis baboon, hamadryas baboon, squirrel monkey, lemur and marmoset. A revised identity of the oblique ligament is proposed as a reinforced distal border of a fibro-osseous annular pollical flexor sheath and whose function is not independent of the flexor sheath. The constant presence and tendinous trait of the pollical oblique ligament in humans, when compared with non-human primates, supports the notion that the oblique ligament strengthens the pollical flexor sheath in humans for restraint of the flexor pollicis longus tendon during forceful precision pinching. A derivation of the pollical oblique ligament is considered as representing a vestigial radial limb of a flexor pollicis superficialis tendon in the thumb. PMID- 12739618 TI - Differential expression of type XII collagen in developing chicken metatarsal tendons. AB - Type XII collagen is a fibril-associated collagen with multiple functional domains. The purpose of this work was to determine its role in regulating tendon matrix assembly. The temporal and spatial expression patterns of both collagen and mRNA were analysed in developing chicken metatarsal tendons using immunofluorescence microscopy, in situ hybridization and real-time quantitative PCR. Temporally, type XII collagen was present during all stages of development (day 14-hatch). However, spatially, type XII collagen expression shifted from the entire tendon at day 14, when the tendon is immature and fascicles are not well developed, to the interfacial matrix (endotendinium) associated with developing fascicles. This shift was obvious beginning at day 17, becoming prominent at day 19. Associated with this shift was a gradual decrease in type XII collagen reactivity in the tendon proper (non-sheath). By hatching, the reactivity was sequestered almost exclusively to the sheaths with some reactivity remaining at the fibroblast-matrix interface within the fascicle. In situ hybridization indicated that fibroblasts in the tendon expressed type XII collagen mRNA homogeneously at day 14. However, by hatching, when the tendon matures, type XII collagen is restricted primarily to the sheath cells. Quantitative PCR analyses, of NC3 splice variants, demonstrated highest expression levels for the short splice variant mRNA at days 14-17, followed by a significant decrease at day 19 with levels remaining constant to adult. Long variant mRNA expression was highest at day 14 then decreased and was constant from day 17 to adult. These changing patterns may be related to the spatial shift in type XII collagen expression to the sheaths. Differential temporal and spatial expression patterns indicate that type XII collagen functions to integrate the developing tendon matrices and fascicles into a functional unit. PMID- 12739619 TI - Intralaryngeal neuroanatomy of the recurrent laryngeal nerve of the rabbit. AB - We undertook this study to determine the detailed neuroanatomy of the terminal branches of the recurrent laryngeal nerve (RLN) in the rabbit to facilitate future neurophysiological recordings from identified branches of this nerve. The whole larynx was isolated post mortem in 17 adult New Zealand White rabbits and prepared using a modified Sihler's technique, which stains axons and renders other tissues transparent so that nerve branches can be seen in whole mount preparations. Of the 34 hemi-laryngeal preparations processed, 28 stained well and these were dissected and used to characterize the neuroanatomy of the RLN. In most cases (23/28) the posterior cricoarytenoid muscle (PCA) was supplied by a single branch arising from the RLN, though in five PCA specimens there were two or three separate branches to the PCA. The interarytenoid muscle (IA) was supplied by two parallel filaments arising from the main trunk of the RLN rostral to the branch(es) to the PCA. The lateral cricoarytenoid muscle (LCA) commonly received innervation from two fine twigs branching from the RLN main trunk and travelling laterally towards the LCA. The remaining fibres of the RLN innervated the thyroarytenoid muscle (TA) and comprised two distinct branches, one supplying the pars vocalis and the other branching extensively to supply the remainder of the TA. No communicating anastomosis between the RLN and superior laryngeal nerve within the larynx was found. Our results suggest it is feasible to make electrophysiological recordings from identified terminal branches of the RLN supplying laryngeal adductor muscles separate from the branch or branches to the PCA. However, the very small size of the motor nerves to the IA and LCA suggests that it would be very difficult to record selectively from the nerve supply to individual laryngeal adductor muscles. PMID- 12739620 TI - Structure of the pelvic and penile urethra--relationship with the ducts of the sex accessory glands of the Mongolian gerbil (Meriones unguiculatus). AB - The urethra is the main place of entry for sexually transmitted pathogens. However, there is little literature on the morphology of the urogenital system, principally the urethra and ducts of the sex accessory glands. The Mongolian gerbil is an insectivorous, herbivorous and monogamous rodent with nocturnal habits; it has been used successfully as a laboratory animal since the 1960s. Therefore, the objective of the present paper was to describe the structure and ultrastructure of the urethra and its relations to the ducts of the accessory sex glands of the Mongolian gerbil (Meriones unguiculatus), contributing to the understanding of the reproductive biology of the rodent and aiming to provide data for future experimental studies. Conventional techniques of light and scanning electron microscopy were utilized. The urethra and ducts of the accessory sex glands are similar to those of the albino rat and the mouse. However, there is variation in drainage type among accessory sex glands for the inner urethra. The ducts of the seminal vesicle, the ductus deferens, drain their contents independently into the ampullary duct that opens in the urethra. The ducts of the prostate, coagulating and bulbourethral glands drain their contents independently into the urethra. PMID- 12739621 TI - Effects of sex hormones on the development of giant lysosomes in the proximal tubules of DBA/2Cr mouse kidney. AB - The DBA/2Cr mouse strain is characterized by giant lysosomes that are located in the proximal convoluted tubules of males and in the proximal straight tubules of females. In the present study, we used light microscopy and electron microscopy to investigate the effects of sex hormones on the development of these giant lysosomes. In the proximal convoluted tubules of males, giant lysosomes (large vacuolar structures observed under light microscopy) disappeared after orchiectomy but reappeared after testosterone treatment. No changes were observed after ovariectomy or estradiol treatment. In the proximal straight tubules of females, giant lysosomes (periodic acid Schiff-positive giant granules observed under light microscopy) disappeared after ovariectomy but reappeared after estradiol treatment. After orchiectomy and testosterone treatment, the giant lysosomes did not appear. However, a number of small lysosomes (smaller than a nucleus), which were periodic acid Schiff-positive, increased after orchiectomy and decreased after testosterone treatment. These findings suggest that lysosomes in DBA/2Cr mice are regulated by testosterone or estradiol as follows: (1) in the proximal convoluted tubules, the development of lysosomes is stimulated by testosterone but not by estradiol; (2) in the proximal straight tubules, development of lysosomes is stimulated by estradiol and inhibited by testosterone. PMID- 12739622 TI - The number of cells expressing dopamine D2 receptor mRNA in rat brain caudate putamen is higher in oestrus. AB - Dopamine D2 receptors (D2-Rs) in the central nervous system are involved in the control of locomotion, cognition, emotion and neuroendocrine secretion. The intensity of cellular responses to specific stimuli is dependent on the concentration of dopamine or its agonist, and the availability, as well as the concentration, of all the other components of the signalling pathway in the cell, including the receptors. Many factors can influence the level of mRNA encoding the receptors. In order to study the changes in the level of expression of the D2 R mRNA in the brain of female rats at different stages of the oestrous cycle, we used a quantitative in situ hybridization technique. Four groups of animals were analysed: rats in prooestrus (POE), oestrus (OE), dioestrus 1 (DOE1) and dioestrus 2 (DOE2). A 35S-labelled riboprobe was transcribed in vitro from the 1.5-kb D2-R cDNA. The caudate putamen of the rats, which shows the highest level of expression of D2-R mRNA in the brain, was examined. The number of silver grains per cell, representing hybridization of riboprobe, and the number of cells expressing the D2-R mRNA, were counted with the computer-assisted image analysis system Lucia-M. Our results show that the expression of the D2-R mRNA in the lateral striatum varies during the oestrous cycle, with the highest expression measured during DOE2. The number of cells expressing the D2-R mRNA also changes during the different phases, with the highest number being detected in OE. This indicates that during OE more cells transcribe the D2-R mRNA. The results suggest that the variations in the concentration of the D2-R mRNA in the caudate putamen of the rat brain at different stages of the reproductive cycle are caused by the combination of variable expression of the mRNA per cell and different number of the cells that express the mRNA. PMID- 12739623 TI - Radiography of soft tissue of the foot and ankle with diffraction enhanced imaging. AB - Non-calcified tissues, including tendons, ligaments, adipose tissue and cartilage, are not visible, for any practical purposes, with conventional X-ray imaging. Therefore, any pathological changes in these tissues generally necessitate detection through magnetic resonance imaging or ultrasound technology. Until recently the development of an X-ray imaging technique that could detect both bone and soft tissues seemed unrealistic. However, the introduction of diffraction enhanced X-ray imaging (DEI) which is capable of rendering images with absorption, refraction and scatter rejection qualities has allowed detection of specific soft tissues based on small differences in tissue densities. Here we show for the first time that DEI allows high contrast imaging of soft tissues, including ligaments, tendons and adipose tissue, of the human foot and ankle. PMID- 12739624 TI - Preservation of three-dimensional capillary structure in frog muscle during aestivation. AB - In mammals, prolonged immobilization of the limbs can result in a loss of capillary tortuosity, resulting in skeletal muscle haemorrhaging if rapid remobilization is permitted. In this study, we examined the effect of 4 months' immobilization on semimembranosus capillary structure in the Green-striped burrowing frog, Cyclorana alboguttata. C. alboguttata routinely aestivates as part of a physiological strategy to avoid desiccation in semi-arid environments and, in this capacity, the hindlimbs of C. alboguttata are immobilized in a cocoon for months at a time. We found that 4 months' aestivation had no effect on three-dimensional capillary structure in the semimembranosus muscle and that capillary tortuosity is preserved in immobilized C. alboguttata. The preservation of capillary structure in the hindlimb muscles of C. alboguttata in part accounts for their remarkable ability to emerge with a fully competent locomotor system after prolonged immobilization. PMID- 12739626 TI - Bringing inorganic chemistry to life. PMID- 12739625 TI - Effects of detailed information about dissection on intentions to bequeath bodies for use in teaching and research. AB - Almost all UK medical schools teach anatomy using human bodies that have been bequeathed specifically for the purpose. Persons intending to make this most generous gift should be fully informed about how their body will be used. If detailed descriptions of dissection reduce the number of bequests, then this traditional and effective approach to anatomy teaching will have to change. To determine what effect detailed information has on intentions to bequeath, the Department of Anatomy at Bristol University sent all 139 people who asked for information, between July and December 2001, a description of dissection that included the statement 'Anatomical examination requires that bodies be dissected (taken apart) so that the fine detail of internal structures can be seen. Organs, such as the heart, lungs and brain, are often removed from the body to allow for more detailed study.' Views were sought by questionnaire (response rate 88%). Ninety-nine per cent of respondents intended to bequeath their body and 88% would allow it to be used in research or teaching with the department to make the final decision. Thus the provision of detailed information about dissection should not reduce the number of bequests and this mechanism could, subject to law, make bodies available for research as well as for teaching. PMID- 12739627 TI - Shape-persistent arylene ethynylene macrocycles: syntheses and supramolecular chemistry. AB - This article describes recent developments in the synthesis of macrocycles having rigid, monocyclic skeletons composed of arylene and ethynylene units and the studies on their self-assembling behavior. PMID- 12739628 TI - Octanuclearity and tetradecanuclearity in manganese chemistry: an octanuclear manganese(II)/(III) complex featuring the novel [Mn8(mu4-O)2(mu3-OH)2]14+ core and [Mn10(II)Mn4(III)O4(O2CMe)20[(2-py)2C(OH)O]4] (2-py = 2-pyridyl). AB - Reactions of Mn sources with di-2-pyridyl ketone, (2-py)2CO, and phenyl 2-pyridyl ketone oxime, (ph)(2-py)CNOH, give the novel clusters [Mn10(II)Mn4(III)O4(O2CMe)20[(2-py)2C(OH)O]4] 1 and [Mn4(II)Mn4(III)O2(OH)2(O2CPh)10[(ph)(2-py)CNO]4] 2, respectively, which possess low-spin ground states; the observed tetradecanuclearity in 1 is extremely rare in 3d-metal chemistry, while the core of 2 has a unique topology consisting of two linked [Mn2(II)Mn2(III)O(OH)] units. PMID- 12739629 TI - Synthesis of protein-nucleic acid conjugates by expressed protein ligation. AB - The synthesis of covalent conjugates of proteins and polyamide nucleic acids (PNA) is accomplished by expressed protein ligation of intein-fusion proteins and a PNA-cysteine conjugate. PMID- 12739631 TI - The structure of a self-assembled calixarene aqua-channel system. AB - The crystal structure of the complex 12.calix-[4]-arene dihydroxyphosphonic acid, 12.propane diammonium, 12.ethanol and 40.water molecules is based on dimeric units of the calix, assembled via trigonal units into a hexameric tube of 15 A radius and 16 A depth, further assemby via spanning propane diammonium cations and ethanol molecules forms a channel (40 A), selectively containing all the water molecules. PMID- 12739632 TI - Adsorbed water for the electro-oxidation of methanol at Pt-Ru alloy. AB - Adsorbed water molecules which promote the methanol oxidation reaction (MOR) at Pt-Ru alloy electrode are clearly detected by in-situ FTIR spectroscopy with the attenuated total reflection configuration, which directly supports the "bi functional mechanism" for the MOR. PMID- 12739630 TI - Isolation of the first ferromagnetically coupled Mn(III/IV) complex. AB - Binuclear manganese complexes Mn2(III/IV)(dtsalpn)2DCBI, 1, Mn2(III/III)(dtsalpn)2HDCBI, 2, containing the ligand dicarboxyimidazole (DCBI) have been prepared in order to address the issue of imidazole bridged and ferromagnetically coupled Mn sites in high oxidation states of the OEC in Photosystem II (PS II). Temperature dependent magnetic susceptibility studies of 1 indicates that the interaction between the two Mn(III)/Mn(IV) ions is ferromagnetic (J = +1.4 cm(-1)). Variable temperature EPR spectra of 1 shows that a g = 2 multiline is as an excited state signal corresponding to S = 1/2. PMID- 12739633 TI - A 3D metal-organic network, [Cu2(glutarate)2(4,4'-bipyridine)], that exhibits single-crystal to single-crystal dehydration and rehydration. AB - The reaction of Cu(NO3)2, glutaric acid and 4,4'-bipyridine in water affords a novel 3D coordination polymer which exhibits reversible desorption and adsorption of water molecules with retention of single crystallinity. PMID- 12739634 TI - A reductive recycle strategy for the facile synthesis of molybdenum(VI) alkylidyne catalysts for alkyne metathesis. AB - A convenient synthesis of trisamido molybdenum(VI) alkylidyne complexes has been developed, in which the key step is the addition of a geminal dichloride to a trisamido molybdenum(III) complex in the presence of magnesium to continuously recycle unwanted side product 4, selectively generating the desired alkylidyne complexes in high yield. PMID- 12739635 TI - The role of the counteranion in the cation-pi interaction. AB - Chemical double mutant cycles have been used to quantify cation-pi interactions in chloroform as a function of the nature of the counteranion. The cation-pi interaction is -2.5 +/- 0.4 kJ mol(-1) and independent of the anion, even though the overall stability of the complexes varies by an order of magnitude due to competition of the anion for alternative binding sites. PMID- 12739636 TI - ZnS-Zn nanocables and ZnS nanotubes. AB - ZnS-Zn nanocables and ZnS nanotubes have been synthesized by a thermochemical process in a simple and safe way. The as-prepared nanocables consist of a single crystal Zn core with a diameter of 20 nm and a polycrystalline ZnS sheath with a thickness of 8 nm. The evaporation of the Zn core leads to the formation of ZnS nanotubes. PMID- 12739638 TI - Unexpected coupling between an eta5-indenyl ligand and alkenyl-vinylidene fragments: synthesis of unprecedented (eta6-indene)ruthenium(II) metallacycles. AB - Vinylidene complexes [Ru[=C=C(H)CR1R2CH2C(Me)=CH2](eta5-C9H7)(PPh3)2][BF4] undergo an intramolecular coupling between the alkenyl-vinylidene fragment and the eta5-indenyl ligand to afford indene-metallacyclic compounds (6a,b) in which the resulting functionalised indene group is eta6-coordinated to the metal. PMID- 12739637 TI - Total synthesis of (+/-)-dihydrospiniferin-1 via a polyfluoro alkanosulfonyl fluoride induced tandem carbonium ion rearrangement reaction. AB - A novel polyfluoroalkanosulfonyl fluoride induced carbonium ion rearrangement reaction of gamma-hydroxymethyl cyclohexenone has been used for the total synthesis of (+/-)dihydrospiniferin 1. PMID- 12739639 TI - Characterization of sulfur exchange reaction between polysulfides and elemental sulfur using a 35S radioisotope tracer method. AB - A sulfur exchange reaction between di-tert-butylpolysulfides and elemental sulfur was examined using a 35S tracer method and the reaction mechanism was discussed. PMID- 12739640 TI - An efficient synthesis of new fluorinated uracil derivatives. AB - A series of potentially biologically active fluorinated uracil derivatives has been prepared in three steps from oxazolines and fluorinated nitriles with good chemical yields. PMID- 12739641 TI - A mono-TTF-annulated porphyrin as a fluorescence switch. AB - Annulation of one TTF unit directly to the porphyrin chromophore produces an almost nonfluorescent species which can be transformed into a fluorescent species by oxidation of the TTF unit. PMID- 12739642 TI - A superior catalyst for low-temperature NO reduction with NH3. AB - Mn-Ce mixed-oxide catalyst yields nearly 100% NO conversion at 100-150 degrees C at a high space velocity of 42,000 h(-1). SO2 and H2O (at high concentrations) have only slight effects on the activity. PMID- 12739643 TI - Why [CpW(CO)3]+ reduces H2 to dihydride. AB - The [CpW(CO)3]+ complex, with three pi acceptor ligands and a positive charge, is shown to have an unexpected reducing ability towards H2 because of a low lying triplet state energy. PMID- 12739644 TI - Palladium-catalysed dimerization of vinylarenes using indium triflate as an effective co-catalyst. AB - A palladium-indium triflate catalyst was found to be much more active for the dimerization of vinylarenes compared with generally used cationic palladium(II) catalysts. PMID- 12739645 TI - RPM-2: a recyclable porous material with unusual adsorption capability: self assembly via structural transformations. AB - Structural transformation via deliberate and partial topological changes in a grid network structure has led to a three-dimensional, fully recyclable porous material (RPM-2) with a very high sorption capability. PMID- 12739646 TI - Dilithiated phosphazenes: scaffolds for the synthesis of olefins through a new class of bicyclic 1,2-oxaphosphetanes. AB - The first examples of the PN-directed dilithiation of (N methoxycarbonyl)phosphazenes in the C(alpha) and C(ortho) to the phosphorus, and the use of these dianions in the formation of tri- and tetra-substituted olefins through stereospecific thermolysis of a new type of isolable bicyclic 1,2 oxaphosphetanes are described. PMID- 12739647 TI - Accelerated Bergman cyclization of porphyrinic-enediynes. AB - The Bergman cyclization of simple diethynylporphyrinic-enediynes exhibits a double activation barrier to the formation of Bergman cyclized product. Addition of H-atom acceptor accelerates the formation of the picenoporphyrin, indicating that the second barrier is rate limiting. PMID- 12739648 TI - Synthesis of transparent and ordered mesoporous silica monolithic films embedded with monomeric zinc phthalocyanine dye. AB - Monolithic transparent mesoporous silica films embedded with zinc phthalocyanine (ZnPc) have been synthesized and it is shown that the encapsulated ZnPc dye molecules exist predominantly in monomeric form. PMID- 12739649 TI - First example of a Sn-C bond cleaved product in the reaction of Ph3SnOSnPh3 with carboxylic acids. 3D-supramolecular network formation in the X-ray crystal structure of [Ph2Sn(OH)OC(O)(Rf)]2, Rf = 2,4,6-(CF3)3C6H2. AB - A 1:2 reaction of Ph3SnOSnPh3 1 with RfCOOH 2 leads to the formation of [Ph2Sn(OH)OC(O)(Rf)]2 3, by means of a facile Sn-C bond cleavage process. PMID- 12739650 TI - Alternating stereospecific copolymerization of cyclopentene and ethylene with constrained geometry catalysts. AB - The stereoselective copolymerization of cyclopentene (cP) and ethylene (E) to generate highly alternating polymers with isotactic cis 1,2-cyclopentene enchainment is reported. PMID- 12739651 TI - Determination of binding affinities on solid supports: influence of the loading and the nature of the solid support. AB - The scope of a solid phase binding assay for the determination of binding affinities between a solid supported substrate and a coloured host has been studied by investigating the influence of the nature and the loading of the solid support. PMID- 12739652 TI - First example of selective hydrogenation of unconstrained alpha,beta-unsaturated ketone to alpha,beta-unsaturated alcohol by molecular hydrogen. PMID- 12739653 TI - Deuterium isotope effect on the solid-state thermal isomerization of photo coloured cis-keto species of N-salicylideneaniline. AB - Deuterium isotope kinetic effect in the solid state was observed by using thermal fading reaction of photo-coloured species derived from N-salicylideneaniline deuterohydroxyl derivative; thus, the existence of a cis-keto form in the photo coloured Schiff base crystals was suggested experimentally along with 6-methyl substitution effect on the stability of the photoproduct. PMID- 12739654 TI - Microporous niobium phosphates and catalytic properties prepared by a supramolecular templating mechanism. AB - Microporous hexagonal niobium phosphate synthesized using neutral surfactants of molecular length, C6 to C10 hydrocarbons, by a supramolecular templating mechanism (S0I0) possesses strong hydrophilic character, which leads to high selectivity for catechol formation (95.3%) in the presence of protic solvent (MeOH) in the hydroxylation of phenol using aqueous H2O2. PMID- 12739655 TI - Hydrogen-bonding based multilayer assemblies by self-deposition of dendrimer. AB - We reported on hydrogen-bonding directed Layer-by-Layer assemblies by self deposition of a kind of dendrimer bearing carboxyl groups on its periphery that act as hydrogen bonding donor as well as hydrogen bonding acceptor. PMID- 12739656 TI - An unprecedented alpha-C-C agostic interaction in a cyclopropyl tris(pyrazolyl)boratoniobium complex. AB - Structural, spectroscopic and theoretical evidence indicate that an unusual alpha C-C agostic interaction is preferred over both alpha- and beta-C-H agostic alternatives in the title compound, TpMe2NbCl(c-C3H5)(MeCCMe). PMID- 12739657 TI - High performance carbon-supported catalysts for fuel cells via phosphonation. AB - Carbon-supported catalysts were phosphonated using 2-aminoethylphosphonic acid, and the resulting catalysts with largely enhanced proton conductivity performed substantially better than the untreated counterparts in proton-exchange membrane fuel cells. PMID- 12739658 TI - A novel imidazolate-bridged heterodinuclear Cu(II)Zn(II) complex derived from a unique macrocyclic ligand with two hydroxyethyl pendants. AB - [(CuimZnL-2H)(CuimZnL-H)](ClO4)3, the first imidazolate-bridged Cu(II)-Zn(II) complex of a unique single macrocyclic ligand with two flexible hydroxyethyl pendants, L (L = 3,6,9,16,19,22-hexaaza-6,19-bis(2 hydroxyethyl)tricyclo[22.2.2.2(11,14)]triaconta-1,11,13,24,27,29-hexaene) has been obtained, in which the macrocyclic ligand with two hydroxyethyl arms possesses a markedly different conformation compared to its dicopper analogue. PMID- 12739659 TI - Synthesis and structure of a new layered zinc phosphite (C5H6N2)Zn(HPO3) containing helical chains. AB - A new compound (C5H6N2)Zn(HPO3) has been prepared hydrothermally; it consists of left-handed and right-handed helical chains that are connected through oxygen atoms to form an undulated sheet structure with 4.8-net. PMID- 12739660 TI - Solid state NMR study of acid sites formed by adsorption of SO3 onto gamma-Al2O3. AB - Detailed structure of Bronsted acid sites on the surface of SO3/Al2O3 catalyst has been proposed based on 1H/27Al TRAPDOR NMR results and the acidity of the catalyst has also been characterized by NMR probe molecules. PMID- 12739661 TI - Ab initio structure study from in-house powder diffraction of a novel ZnS(EN)0.5 structure with layered wurtzite ZnS fragment. AB - The solvothermal reaction of elemental zinc with sulfur in ethylenediamine (en) as solvent yields [ZnS-0.5(NH2CH2CH2NH2)], 1, an unprecedented ethylenediamine pillared ZnS layered compound, containing two dimensional (2-D) boat-type 6 membered rings, which was characterized by ab initio structure solution from powder diffraction data (SDPD). PMID- 12739662 TI - Self-assembly of a ferrocene-substituted porphyrin capable of electrochemically sensing neutral molecules via a "tail on-tail off" process. AB - The supramolecular assembly of a ferrocene-porphyrin conjugate allowed ferrocene based electrochemical sensing of the metalloporphyrin axial coordination via a "tail on-tail off" binding process. PMID- 12739663 TI - 8-(1,4,7,10-Tetraoxa-13-azacyclopentadec-13-ylmethyl)quinolin-7-ol: synthesis and application as a highly sensitive metal cation probe. AB - A new metal ion probe 8-(1,4,7,10-tetraoxa-13-azacyclopentadec-13 ylmethyl)quinolin-7-ol (1a) was synthesized via a modified Mannich reaction, in which the mechanism of recognition incorporates excited-state proton transfer reactions. The remarkable differentiation in spectral properties upon metal complexation makes 1a a highly sensitive fluorescence probe. PMID- 12739664 TI - Novel economic and green approach to the synthesis of highly active W-MCM41 catalyst in oxidative cleavage of cyclopentene. AB - A highly active and perfectly structured W-MCM41 catalyst for the oxidative cleavage of cyclopentene to glutaraldehyde was synthesized through a novel economic and green synthetic method by using Na2SiO3 as the Si source and CH3COOC2H5 as the hydrolyzer. PMID- 12739665 TI - Perylenediimide-surfactant complexes: thermotropic liquid-crystalline materials via ionic self-assembly. AB - In this communication we present the facile preparation and characterisation of thermotropic liquid-crystalline materials from the ionic self-assembly of a charged perylenediimide derivative and oppositely charged surfactants. PMID- 12739666 TI - A rapid and effective synthesis of propylene carbonate using a supercritical CO2 ionic liquid system. AB - The synthesis of propylene carbonate from propylene oxide and carbon dioxide under supercritical conditions in the presence of 1-octyl-3-methylimidazolium tetrafluoroborate was achieved in nearly 100% yield and 100% selectivity within 5 minutes, whose TOF value is 77 times larger than those so far reported. PMID- 12739667 TI - New synthetic route for organic polyoxometallic clusters: synthetic and structural investigations on the first dumb-bell shaped polyoxozirconium hydroxide with the [Zr9(mu5-O)2(mu3-O)4(mu-O)4(mu-OH)8] core structure. AB - A new route for organic polyoxometallic clusters describes the first dumb-bell like organic polyoxozirconium hydroxide [[(Cp*Zr)4(mu5-O)(mu3-O)2(mu-OH)4]2Zr(mu O)4] x 2C7H8 (2; Cp* = C5Me5) involving the treatment of the Bronsted acidic organozirconium hydroxide [(Cp*Zr)6(mu4-O)(mu-O)4(mu-OH)8] x 2C7H8 (1) with organozirconium compounds. PMID- 12739668 TI - Chemical modification of diamond powder using photolysis of perfluoroazooctane. AB - Photolysis of perfluoroazooctane with diamond powders led to chemical modification of the surface with the introduction of perfluorooctyl ester and ether functional groups, the presence of which was confirmed by means of FT-IR, XPS and 19F NMR measurements. PMID- 12739669 TI - Large size anion binding with iron(II) complexes of a 5,5'-disubstituted-2,2' bipyridine ligand. AB - Three 5,5'-substituted 2,2'-bipyridyl ligands around a metal atom form two clefts which can encapsulate sulfate, perchlorate, or nitrate anions. PMID- 12739670 TI - A novel approach to carbon hollow spheres and vessels from CCl4 at low temperatures. AB - Carbon hollow spheres (400-600 nm) and vessels (400 nm x 3000 nm) have been synthesized from sp3-CCl4 at 190 and 230 degrees C, respectively. The HRTEM images and Raman spectra reveal the sp2 nature of the as-obtained products, indicating that the transformation from carbon sp3 to sp2 occurs in the reactions. The possible mechanism has also been proposed. PMID- 12739671 TI - Synthesis, separation, and isomer-dependent packing in two dimensions--detected by scanning tunnelling microscopy--of a TTF derivative. AB - The synthesis, isolation and STM imaging on graphite of the cis and trans isomers of a TTF reveal isomer-dependent packing, and constitutes a way to study the non covalent interactions at play in these systems. PMID- 12739672 TI - CuCl catalyst heterogenized on diamide immobilized SBA-15 for efficient oxidative carbonylation of methanol to dimethylcarbonate. AB - CuCl has been successfully immobilized on a novel diamide modified SBA-15, and proven to be an efficient heterogenized catalyst for the oxidative carbonylation of methanol to dimethylcarbonate. PMID- 12739674 TI - Covalent immobilization of oligonucleotides on p-aminophenyl-modified carbon screen-printed electrodes for viral DNA sensing. AB - DNA-sensing platforms were prepared by covalently attaching oligonucleotide capture probes onto p-aminophenyl-functionalized carbon surfaces and applied to the determination of an amplified herpes virus DNA sequence in an electrochemical hybridization assay. PMID- 12739675 TI - Reversible regulation of pyrene excimer emission by light and metal ions in the presence of photochromic spiropyran: toward creation of a new molecular logic circuit. AB - The excimer fluorescence of a bis-pyrene molecule can be reversibly regulated by ultraviolet light, metal ions and visible light in the presence of spiropyran. Based on this result, a new molecular logic circuit is proposed. PMID- 12739673 TI - Luminescence study of tetravalent uranium in aqueous solution. AB - The luminescence spectrum of U4+ in aqueous solution was observed in the UV-Vis region with the lifetime < 20 ns at room temperature by excitation light corresponding to the 5f-5f electronic transition. All the luminescence peaks were assigned to individual electronic transitions. PMID- 12739676 TI - Highly efficient Lewis acid-catalysed Pictet-Spengler reactions discovered by parallel screening. AB - High yielding Lewis acid-catalysed one-pot Pictet-Spengler reactions of tryptophan methyl ester and tryptamine with aliphatic and aromatic aldehydes were achieved in short reaction times with the aid of microwave irradiation. PMID- 12739677 TI - TMJ fractures in children and adolescents: treatment guidelines. AB - TMJ fractures must be focused not only as a cause of direct damage to osseous structures, but also of future disturbances of dentofacial development. Treatment is aimed at restoring normal joint function, occlusion and symmetry. Any disturbance of condylar cartilage will result in alteration of mandibular development. The subsequent deformity of jaw and face will depend not only on the type, intensity extent and chronology of the noxious agent, but also on the particular time of occurrence and growth activity. Thus the effect will be more decided if the disturbance occurs early in life, during childhood, when growth activity is greater and mandibular shape and size have not been assumed yet. This report will include basic information on both prevention and first aid in these types of injuries. The correct application of these precautions immediately following the trauma should improve the short and long-term outcome. Information on diagnosis and treatment of lesions of the bone and soft tissues, which may coexist with dental trauma, a critical first step in the overall management of traumatized patients, will be given. Follow-up procedures of these patients will be illustrated. The guidelines described in this paper for the treatment of traumatic TMJ fractures in children and youths are based on our clinical experience. They are intended as an aid to practioners in the management and treatment of these traumas, by professional must always use professional judgement. There are no guarantees of any positive results associated with the use of these guidelines, although it is felt that time and proper treatment will maximize the chances of success. PMID- 12739679 TI - History of water fluoridation. AB - Fluoride, the milestone element of pediatric dentistry, as well as modern dentistry, has been utilized for the last six decades in practice. Researchers all over the world have used fluoride in several forms in the prevention of tooth decay. Today, we are aware that most of this work has involved clinical research with children. It is particularly pertinent that the subject be adequately presented in the field of pediatric dentistry. In 1940's and 1950's were the years of ten-year studies, where caries reductions were first described. The industry of systemic fluoridation, public campaigns and advertisements became popular mostly in western world in 1960's and 1970's. The fluoridated dentifrices and changes of dentifrice formulations were established in 1980's. Many stated that children should receive one form of systemic fluoride and appropriate forms of topical fluoride in 1990's. Analyses showed that maximum protection against caries is obtained when teeth erupt into an environment with low concentrations of ionic fluoride. The similarity in caries reductions obtained in water fluoridation studies and long-term studies with topically administered fluoride regimens, including fluoride-containing dentifrices, indicates that the pre eruptive effect of fluoride is of borderline significance relative to the more significant post-eruptive effect. It has taken a long time to show that water fluoridation and topical fluoride programs were thus important measures for the control of caries at the community level. Today, we are aware of the fact that the trio of diet, dentifrice and supplementation should exceed the optimal levels. PMID- 12739678 TI - Class IV preparations for fractured anterior teeth restored with composite resin restorations. AB - Pediatric dentists play a major role in treating most of the anterior teeth fractures due to the fact that most patients who suffer such traumatic injuries are between the ages of seven and fourteen. Several techniques has been developed to restore the fractured incisors to the original shape and color, these include full-coverage of the traumatized tooth, or restoring the incisors with a resin. The purpose of this study is to find the ideal combination of tooth preparation and restorative material, namely, to determine if stair step chamfer preparations provide more retention in class IV restorations than the plain chamfer preparation technique when restored with a combination of a hybrid composite resin and a microfilled composite resin. This was done by comparing the shear strength values between the buccal stair-step chamfer preparation, and a modification on it (buccal and lingual stair-step chamfer preparation) and the plain chamfer preparation techniques in class IV restorations on anterior incisors using different composite resin materials. The Instron machine was used to test shear strength. One hundred and two bovine incisor teeth were freshly harvested from the slaughterhouse. The teeth were prepared and restored according to the following six groups; Plain Chamfer preparation with Tetric Ceram Composite, Plain Chamfer preparation with Renamel Composite, buccal stair-step chamfer preparation with Tetric Ceram composite, buccal stair-step chamfer preparation with Renamel composite, Buccal and lingual stair-step preparation with Tetric Ceram Composite, Buccal and lingual stair-step chamfer preparation with renamel composite. All samples were fractured using the Instron testing machine then the surface area were measured using Image-J software. Shear strength for every sample was calculated using the load numeric result from the Instron machine and the measured surface area. The Two-Factorial (AB) Analysis of Variance For Independent Samples showed that the buccal stair-step chamfer preparation showed significantly higher shear strength and fracture resistance than plain chamfer or the buccal and lingual stair-step chamfer preparation. The combination of Renamel Hybrid and Renamel Microfill composite materials showed better results than the Tetric Ceram composite when used with all three preparation techniques, but did not show a statistical significance. It was concluded that buccal stair-step preparation technique provided the ideal preparation technique with bonded composite resins in fractured anterior teeth. Only 7% of the entire sample size had an adhesion failure versus 93% that had cohesion failure. There was no significant difference in shear strength, between the restorative materials used, in conjunction with all the preparation techniques. PMID- 12739680 TI - The use of overdentures in the management of severe hypodontia associated with microdontia: a case report. AB - Dental anomalies may compromise both esthetics and function in the pediatric patient. The present paper reports a rare case associated with both microdontia and severe hypodontia in a 13 year-old girl. The treatment was accomplished through the use of overdentures, which also served as a bite plane to increase vertical dimension of occlusion and as a potential space maintainer. PMID- 12739681 TI - Permanent incisors traumatized through predecessors: sequelae and possible management. AB - This report describes the sequence of events following a traumatic dental injury to primary maxillary anterior teeth of a five-year old male during a bicycle accident. The development of the affected teeth during the subsequent ten years was studied from the radiographic records. A detailed account of the clinical and radiographic findings as well as the physio-pathologic changes that have taken place is discussed. In view of the inevitable prognosis of dentitions that sustain traumatic injury during the formative years preventive measures and feasible dental management are proposed. These precautionary measures are aimed to circumvent the sequelae that were experienced in the present case, and avoid predictable complications. PMID- 12739682 TI - Orthodontic treatment of a case of Aarskog syndrome. AB - Aarskog syndrome is a rare syndrome with a typical triad of facial, digital and genital characteristics. Orthodontic treatment of a patient with Aarskog syndrome has not been documented in literature till date. This case report shows that an individual with Aarskog syndrome can be an excellent orthodontic patient and should not be excluded from the general patient population. A stable occlusion can be achieved with acceptable profile and soft tissue contours in these patients as well. PMID- 12739683 TI - Healing of root resorption: a case report. AB - External resorption is sequelae of necrotic periodontal membrane over a large area of root following an injury to the tooth. This usually occurs after severe dental injuries such as intrusion, severe luxations or exarticulation injuries complicated by a prolonged extra oral period. This case report presents a clinical and radiographic follow up (13 months) of treatment of inflammatory external root resorption on maxillary central incisor using Vitapex. Gradual healing of resorption was observed radiographically with no tenderness or pathological mobility. PMID- 12739684 TI - Failure of eruption of first and second permanent molars. AB - Failure of eruption of permanent molars is an uncommon condition with a range of possible causative factors. This retrospective study of 35 pediatric subjects assesses the influence of these factors in the prognosis. The study aims to broaden our understanding of an abnormality, which has a considerable clinical impact, and proposes a treatment protocol for the management of these patients. PMID- 12739685 TI - Assessing the pain reaction of children receiving periodontal ligament anesthesia using a computerized device (Wand). AB - The purpose of the study was to compare the reaction of children, who received a local anesthesia to the upper incisors with a conventional buccal infiltration and a periodontal ligament injection (PDLi) with a computerized device (Wand); to assess the efficacy of the anesthesia and the reactions by children after treatment. Ninety-eight children aged 2 to 4 years participated in the study. All the children were sedated with hydroxyzine and nitrous oxide. Children were randomly assigned to receive either PDLi with the Wand or conventional infiltration. During the injection, a modified Behavioral Pain Scale was used. There were 21 girls and 23 boys between the ages of 2 to 4 years in the study group (mean age 3.9 +/- 1.3 years), and 16 girls and 29 boys (mean age 3.5 +/- 1.0 years) in the control group. No significant difference was found between boys and girls. More children reacted negatively, namely, crying, facial expressions and eyes squeezed, while receiving the conventional infiltrative injection, whereas children, who received the anesthetic solution using the Wand, reacted more positively. This difference was statistically significant. No significant difference regarding the efficacy of the anesthesia was observed in either technique. Eighty percent of the children who receive the conventional buccal infiltration scratched the nose or upper lip after the treatment, while none of the children, who received PDLi, showed signs of discomfort. Children displayed better behavior when they received local anesthesia with the Wand rather than with the conventional infiltration. The same efficacy of anesthesia was achieved with both techniques. Children did not show signs of discomfort after treatment with the Wand, whereas they did while receiving conventional injections. PMID- 12739686 TI - Antibacterial and conventional self-etching primer system: morphological evaluation of intact primary enamel. AB - The aim of this study was to analyze the etching effect of a new dental adhesive system that uses an antibacterial and self-etching primer agent. Non-carious primary human molars were divided into three groups: 35% phosphoric acid etching (3M-ESPE), self-etching primer (Clearfil SE Bond--Kuraray) and antibacterial self etching primer (ABF experimental system--Kuraray) for SEM evaluation of surface, resin replicas and interface. Both self-etching primers produced shallow grooves and no defined etching morphologies. Phosphoric acid produced definite etching patterns at the enamel surface. PMID- 12739687 TI - Estimation of the fluoride concentrations in human breast milk, cow's milk and infant formulae. AB - Fluoride has a significant effect on the prevention of dental caries. The major dietary intake of the infants constitutes the breast milk cow's milk and infant milk formulations in which the fluoride contents varies widely. Hence it is important to identify the potential milk source of high fluoride intake in an infant's diet and to evaluate the need for the fluoride supplementation. The aim of the present study was to determine the fluoride concentrations in breast milk, cow's milk, infant formulae and water samples in Mangalore city, India, using fluoride electrode (Orion Model 940900). It was noted that minimal amount of fluoride was found to be present in breast milk and cow's milk samples, while the fluoride content of water samples was found to be below the optimum level. On the contrary the infant formulae were found to have equal or more levels of fluoride. PMID- 12739688 TI - Aspects of mandibular morphology, with specific reference to the antegonial notch and the curve of Spee. AB - The purpose of this investigation was to determine whether particular metrical traits of the mandibular antegonial notches are associated with specific mandibular growth patterns, and also with the mean depth of the curve of Spee. Thirty pre-treatment lateral cephalometric radiographs belonging to a randomly selected group of patients treated in the orthodontic clinic at the SUNY at Buffalo were digitized. The surface areas of the mandibular antegonial notches, as well as some sagittal facial dimensions were measured on each radiograph. The curve of Spee was measured directly from the pre-treatment mandibular study cast of each patient included in this study. An analysis of variance showed no statistically significant difference (p < 0.01) between the measurements recorded by the two examiners who conducted this study. The results of this study showed a statistically significant positive correlation between the surface areas of the antegonial notches and the lower anterior facial heights (r = 0.87, P < 0.001). At the same time a statistically significant negative correlation was found between the surface areas of the antegonial notches and the lengths of the mandibular bodies (r = -0.9, P < 0.001). A significant negative statistical relationship was shown to exist between the lower anterior facial heights and the lengths of the corresponding mandibular bodies, and also between the depths of the curves of Spee and the surface area of the respective antegonial notches (r = -0.85, P < 0.002). The results of this study indicate that an increase in the areas of the antegonial notches is associated with a tendency for greater vertical growth of the mandible. The results further suggest that the depth of the curve of Spee and the length of the mandibular body are decreased when there is an increase in the surface areas of the antegonial notches. PMID- 12739689 TI - Variations among the primary maxillary dental arch forms using a polynominal equation model. AB - This study identifies arch symmetry and describes arch form variations in normal primary maxillary dental casts. Sixty-two percent of casts had asymmetrical arches, and 38% had symmetrical arches. Six types of arch form were identified according to the ratio of arch width to length. There were more long types than short types. The rounded and short types tended to have a more-symmetrical arch than did the long type. PMID- 12739690 TI - Leakage reduction with a surface-penetrating sealant around stainless-steel orthodontic brackets bonded with a light cured composite resin. AB - In this study a surface penetrating sealant was used around bonded stainless steel orthodontic brackets. The investigation attempted to identify the effect of surface penetrating sealants on the microleakage associated with orthodontic stainless steel brackets bonded with light cured composite resin. A total of 58 bovine teeth were used for this study, orthodontic brackets were bonded with light cured orthodontic resin. The following groups were assigned: (I) Finished but unsealed, (II) Finished and sealed, and (III) Un-finished but unsealed. The brackets were activated using orthodontic elastics, stained, sectioned, and evaluated under magnification. The following statistical analysis was done Mann Whitney U test for two independent samples, then confirmed with a Kruskal-Wallis One-Way analysis of variance by ranks. The second and third groups were statistically better in respect of marginal integrity as compared to the first. PMID- 12739691 TI - A comparative study of dentin thickness of primary human molars. AB - The aim of the present survey was to evaluate the dentin thickness of buccal, lingual, mesial and distal surfaces of first and second primary molars. These evaluations were done at the middle third of the crowns of twenty extracted non carious primary teeth separated in two groups (first molars and second molars). Comparisons were performed within-groups and between-groups. The mean values of dentin thickness observed for first molars were 2430 mm (buccal), 1.869 mm (lingual), 1.655 mm (mesial) and 1.664 mm (distal). For second molars the dentin thickness presented 3.006 mm (buccal), 2730 mm (lingual), 2130 mm (mesial) and 2192 mm (distal). The dentin thickness was higher in the buccal surface in both groups (P<0.05; Wilcoxon Test), except for the lingual surface of second molars (P>0.05). No differences were observed between mesial and distal surfaces in both groups (P>0.05). Comparisons between-groups showed that the dentin thickness was higher in all surfaces of second molars (P<0.01; Mann-Whitney Test). It was concluded that there are differences in the dentin thickness between first and second primary human molars and within the different surfaces of such teeth. PMID- 12739692 TI - Peripheral ossifying fibroma in a child: report of case. AB - A typical peripheral ossifying fibroma in the anterior maxilla of an 11-year-old boy is presented. The importance of differential diagnosis and proper treatment for prevention of recurrence is discussed. PMID- 12739693 TI - Dental caries prevalence in the primary dentition of cerebral-palsied children. AB - The aim of this study was to evaluate dental caries prevalence in the primary dentition of children with cerebral palsy, as well as to determine some possible risk factors involved in establishing this disease. From this study, it was concluded that children with cerebral palsy had greater prevalence of dental caries in the primary dentition than normal children, and that the evaluated risk factors did not influence this result. PMID- 12739694 TI - Dental caries status of children with bronchial asthma. AB - The purpose of the present study was to determine the dental caries status in children with bronchial asthma and to examine the correlation with the severity of asthma and the form of medication being taken. Two hundred and five asthmatic children of both sexes from three to eighteen years old participated in this study. Children were divided into groups: primary, mixed and permanent dentitions. They were also grouped depending on the severity of asthma into mild, moderate and severe asthma and depending on the form of medication being used into inhaler, syrup, tablet and combination group. In the primary dentition group, the asthmatic children had a mean deft of 5.02 +/- 3.05 and a caries prevalence of 75.75%, in the mixed dentition group, the mean deft was 3.45 +/- 2.89 and mean DMFT was 4.83 +/- 3.66 with a caries prevalence of 78.31%. In the permanent dentition the mean DMFT was 5.17 +/- 4.54 and a caries prevalence of 83.15%. Asthmatic children had a high caries prevalence and it increased with the severity of bronchial asthma. The caries prevalence was highest in those taking medication in the form of syrup. PMID- 12739695 TI - Who is mommy tonight? Lesbian parenting issues. AB - Developing an identity as mother and establishing what it means to mother is a task not defined by sexual orientation. Both heterosexual and homosexual women encounter role models and the dictates of society when mothering (Nelson, 1996). The present study explores how 18 lesbian adoptive parents, 49 lesbian parents who formed their families biologically, and 44 heterosexual adoptive parents experience and perceive their parenting role, how they respond when their children seek them or their partner for particular nurturing, and how the parents negotiate the cultural expectation of a primary caregiver. Lesbian couples were more equal in their division of childcare than heterosexual parents, and lesbian adoptive parents were the most egalitarian. In all types of dual parent families, parents were sought by their child for different activities. In heterosexual adoptive and lesbian biological families, the child's parental preference was rarely a source of conflict between partners. Lesbian adoptive parents were more likely to report that this preference caused occasional conflict. Reasons for this conflict are discussed in light of societal expectations of women and the role of mother. PMID- 12739696 TI - Development and validation of a scale measuring modern prejudice toward gay men and lesbian women. AB - This paper describes the psychometric properties of the Modern Homonegativity Scale (MHS), which measures contemporary negative attitudes toward gay men and lesbians (i.e., attitudes not based on traditional or moral objections to homosexuality). In Study 1 (N = 353), a preliminary version of the MHS was developed, and its psychometric properties were examined. Participants in Studies 2 and 3 (Ns = 308 and 233, respectively) completed the MHS and other attitudinal measures. The relationships among these variables were investigated to provide a more comprehensive assessment of the scale's construct validity. In Study 4 (N = 36), a behavioural expression of modern homonegativity was examined using the attributional ambiguity paradigm. The results of these studies indicate that the MHS is unidimensional, possesses a high degree of internal consistency, and is factorially distinct from a measure of old-fashioned homonegativity. As hypothesized, scores on the MHS correlated positively with political conservatism, religious behaviour, religious self-schema and modern sexism, but did not correlate significantly with social desirability bias. In addition, the MHS appears to be less susceptible to floor effects than a commonly used measure of old-fashioned homonegativity. Finally, the experimental study revealed that participants obtaining high scores on the MHS were less likely to sit beside individuals wearing T-shirts with pro-gay or pro-lesbian slogans when they could justify their seating choice on nonprejudicial grounds. PMID- 12739697 TI - Attribution of rape blame as a function of victim gender and sexuality, and perceived similarity to the victim. AB - This study examined respondents' perceived level of blame and responsibility for three victims of rape, as a function of attitudes toward homosexuals, and perceived similarity to the victim, as indicative of Shaver's (1970) Defensive Attribution Hypothesis. Victims were a homosexual and heterosexual male, and a female. A sample of 168 university students completed questionnaires, which included three rape scenarios and subsequent questions, the Index of Attitudes Toward Homosexuals (Hudson & Ricketts, 1980), and the short-form Marlowe-Crowne Social Desirability Scale (Reynolds, 1982). Results indicated that respondents higher in homophobia (regardless of gender) blamed the homosexual male rape victim and the behavior and character of the heterosexual male rape victim, more than the female rape victim. Male respondents in general also blamed the heterosexual male rape victim, more than female respondents. Shaver's defensive attribution hypothesis was not supported. Results are discussed in terms of the possible link between homophobia and male blame. PMID- 12739698 TI - Crooked men and straightened women: images of homosexuality across race in two women's magazines, 1986-1995. AB - Images of traditional sexuality, across race, prevail in mass media, especially in women's magazines. While omitting less traditional sexual practices for the most part, New Woman and Essence magazines construct the image of a bisexual male who threatens the heterosexual woman and the legitimate heterosexual couple. These women's magazines also redefine homosexual activity, suggesting women who experience homosexual inclinations are really engaging in normal heterosexual behavior. PMID- 12739699 TI - Correlates of internalized homophobia and homosexual identity formation in a sample of gay men. AB - Eighty-six behaviourally homosexual men, at various stages of homosexual identity formation (HIF), were participants in a study that examined correlates of internalized homophobia (IH) and HIF. Results indicated higher levels of internalized homophobia were correlated with lower stages of HIF. In addition, internalized homophobia was significantly related to lower levels of self-esteem, to lower levels of self-concepts of physical appearance and emotional stability, and to higher levels of sex guilt. Post hoc regression analysis revealed that perceived level of repression against homosexuals predicted internalized homophobia and sex-guilt, which were mediator variables to the resolution of HIF. This research supports theory, that suggests internalized homophobia is a major developmental difficulty in gay men. PMID- 12739700 TI - The effect of homosexuality on perceptions of persuasiveness and trustworthiness. AB - This study investigated the effect of sexual orientation on perceptions of persuasiveness and trustworthiness. Subjects were told that a university was considering adding either an International Studies Minor or a Gay, Lesbian, and Bisexual Studies minor to its curriculum. Subjects then viewed a videotape of a male professor, introduced as either homosexual or heterosexual, speaking about the benefits of the proposal. Following the video, ratings of the speaker's trustworthiness and persuasiveness were assessed. Results indicate that the speaker's sexual orientation alone did not have a significant effect on either of these ratings. However, a significant interaction occurred between the speaker's sexual orientation and the speech topic, indicating that subjects expressed especially low levels of trustworthiness when the homosexual speaker was lecturing on the GLB minor. Regression analyses revealed that subjects' attitudes toward homosexuals were a significant predictor of their judgements of the speaker's trustworthiness. PMID- 12739701 TI - Pictures of male homosexuals in the heads of Turkish college students: the effects of sex difference and social contact on stereotyping. AB - The purpose of this study was to focus on the context of stereotypes of gay men, and how sex differences and social contact influence the stereotypic and counter stereotypic attributes of them. To do that, 77 female and 106 male heterosexual undergraduate students participated in the study. The participants filled out a rating scale. The findings of the study demonstrated that male participants defined male homosexuals by using more stereotypic, less counter-stereotypic attributes, while female participants used less stereotypic and more counter stereotypic attributes to define gay men. In the same way, participants who hadn't had any social contact with any homosexuals used more stereotypic and less counter-stereotypic attributes, while others who had had social contact with at least a homosexual defined gay men by using less stereotypic and more counter stereotypic attributes. Both male participants and participants who hadn't had social contact with any homosexuals used more negative stereotypes than both females and participants who at least had had social contact with a homosexual. The findings were discussed in terms of gender-based judgments of gay men. PMID- 12739702 TI - Masculinity and desire in the works of J. R. Ackerley. AB - Although homosexuality was still identified as a sign of effeminacy in the West at the beginning of the twentieth century, the work of George Chauncey suggests that working-class men in the West were able to maintain a masculine identity by playing the active, or insertive, role in sexual encounters. The experiences of one middle-class English homosexual, J. R. Ackerley (1896-1967), reveals that maintaining one's masculinity was more difficult for men of the middle class who desired same-sex contact. After a period in which his identity vacillated between the poles of male and female, he conceived a masculine homosexual identity by consciously rejecting effeminacy in himself and others, by dressing as a "normal" middle-class male, and, most importantly, by seeking relationships with younger, working-class men. PMID- 12739703 TI - New opportunities to study real-world patient management. PMID- 12739704 TI - Effect of loperamide on gastro-oesophageal reflux. AB - BACKGROUND: We have recently shown that morphine reduces gastro-oesophageal reflux (GOR) by stimulating opioid receptors. Our aim was therefore to investigate the effect of loperamide, a peripheral opioid agonist, on GOR. METHODS: Nine patients with GOR disease underwent two ambulatory 24-h intra oesophageal pH monitoring sessions separated by a period of at least 7 days under double-blind randomized conditions during the administration of placebo or loperamide solution p.o. (8 mg + 4 mg + 4 mg, 12 h and 2 h before, and 7 h after the start of the recording). RESULTS: Twenty-four-hour oesophageal acid exposure and the number of reflux episodes/hour were similar under the two experimental conditions: a median (IQ range) of 6.2% (3.3%-11.0%) of the time at pH < 4 and 2.6 (1.6-3.5)/h during placebo versus 8.8% (3.7%-14.8%) and 2.8 (2.3-4.9)/h during loperamide (P = ns). Acid clearance time was also similar: 1.8 +/- 0.3 min versus 1.6 +/- 0.2 min (P = ns). CONCLUSIONS: Loperamide did not decrease GOR, thus suggesting that an opioid agonist may not be useful in the treatment of GOR disease. PMID- 12739705 TI - Symptom-relieving effect of esomeprazole 40 mg daily in patients with heartburn. AB - BACKGROUND: To assess symptom relief in patients with heartburn following treatment with esomeprazole 40 mg daily. METHODS: Patients with heartburn (for > or = 6 months) were assessed in this double-blind, multicenter study. After a 3 day single-blind placebo run-in, 440 patients were randomized to esomeprazole 40 mg o.d., esomeprazole 20 mg b.i.d. or placebo for 14 days. Heartburn symptoms were recorded daily; as insufficient patients had data available from days 13 and 14, analyses included data up to day 12. Gastroesophageal reflux disease (GERD) was diagnosed by upper GI endoscopy and 24-h pH-monitoring. The primary end-point was total heartburn relief defined as no heartburn symptoms during the preceding 24-h period. RESULTS: 240 patients had erosive esophagitis (EO) and 114 patients had GERD defined by pH-monitoring. Proportions of patients with total heartburn relief increased during the first days of treatment and stabilized after Day 4. Total heartburn relief occurred in 67%-73%, 62%-70%, and 21%-32% of patients in the esomeprazole 40 mg o.d., esomeprazole 20 mg b.i.d., and placebo groups, respectively, between days 6 and 12. Proportions of patients with total heartburn relief were higher in patients with EO (71%-80% of patients from Day 4 onwards) compared to those without EO (52%-67% of patients from Day 4 onwards). Figures for patients diagnosed by pH-monitoring were 65%-73% of those with a positive diagnosis and 51%-58% with a negative diagnosis. CONCLUSION: Esomeprazole 40 mg o.d. treatment produces total heartburn relief in a high proportion of patients with GERD. Once-daily esomeprazole 40 mg dosing is recommended as no advantage was gained by splitting the dose. PMID- 12739706 TI - One-week esomeprazole treatment: an effective confirmatory test in patients with suspected gastroesophageal reflux disease. AB - BACKGROUND: Symptomatic response to acid inhibition can be used as a guide in diagnosing patients with reflux symptoms. The proton-pump inhibitor (PPI) omeprazole has been used as such a diagnostic tool. Intragastric acid control with esomeprazole is more effective than other PPIs and has the potential to offer an advance in the diagnostic use of PPIs. METHODS: Patients experiencing heartburn (for > or = 6 months) were studied in this randomized, double-blind, multicenter study. Following a 3-day placebo run-in, 440 patients were randomized to 14 days' treatment with esomeprazole 40 mg once daily (o.d.), esomeprazole 20 mg twice daily (b.i.d.) or placebo. Heartburn symptoms were recorded daily. Endoscopy and 24-h esophageal pH-monitoring were performed to determine the presence of gastroesophageal reflux disease (GERD). The esomeprazole treatment test was considered positive if patients' symptoms improved during the treatment period compared with symptoms recorded on Day 0. RESULTS: 63 patients were excluded from the analysis due to lack of symptoms on Day 0 or failure to complete pH-monitoring. The sensitivity of an esomeprazole treatment test in confirming GERD increased during the first days of treatment and stabilized between 79% and 86% after 5 days (both esomeprazole arms). The corresponding figure for the placebo arm was 36%. Specificity was variable (24%-65%) for both active treatment and placebo. CONCLUSION: A treatment test with esomeprazole 40 mg has a high sensitivity in confirming GERD. Furthermore, the data indicate that 1-week treatment with a once-daily dosage is sufficient to ensure adequate diagnosis. PMID- 12739707 TI - Long-term treatment with sterigmatocystin, a fungus toxin, enhances the development of intestinal metaplasia of gastric mucosa in Helicobacter pylori infected Mongolian gerbils. AB - BACKGROUND: Helicobacter pylori is a human gastric carcinogen. Sterigmatocystin (ST), a fungus toxin, is a risk factor of gastric cancer. Cytotoxin-vacuolation toxin A (VacA) present in supernatants of H. pylori suspensions can cause gastritis and ulcer. The aim of this study was to examine the effects of H. pylori, ST and VacA in Mongolian gerbils. METHODS: Male Mongolian gerbils (n = 196) were treated with H. pylori supernatants (10 ml/1000 mg) mixed with diet or inoculated intragastrically with H. pylori alone or with ST (100 or 1000 ppb), and then killed 27 months later. Gastric tissue sections were stained with haematoxylin and eosin (H&E), periodic acid-Schiff (PAS), Alcian blue (AB, pH 2.5) and with immunostaining for PCNA and p53 expression. RESULTS: In H. pylori infected gerbils, the normal mucosa was replaced by hyperplastic epithelium. Severe gastritis, cystic dilatation of gastric glands, hyperplastic polyps and intestinal metaplasia were observed. In H. pylori + ST (1000 ppb) gerbils, intestinal metaplasia was significantly more frequent than in H. pylori alone animals. No pathological changes were observed in the H. pylori supernatant group. Osseous metaplasia was observed in the H. pylori + ST (100 ppb) group. Serum gastrin levels of the H. pylori + ST (1000 ppb) group were significantly higher than those of the other groups. PCNA labelling index and p53 index of infected gerbils were significantly higher than those of uninfected groups. CONCLUSION: H. pylori causes gastritis, ulcer and intestinal metaplasia. ST enhances the development of intestinal metaplasia and increases gastrin levels in H. pylori-infected Mongolian gerbils. PMID- 12739708 TI - Duodenogastric reflux and Helicobacter pylori infection synergistically increase gastric mucosal cell proliferative activity in Mongolian gerbils. AB - BACKGROUND: Helicobacter pylori and duodenogastric reflux (DGR) are both recognized as aetiological factors in chronic gastritis and gastric carcinogenesis. In this study, a Mongolian gerbil (MG) model was used to investigate the histopathological changes in the gastric mucosa resulting from DGR and/or H. pylori infection. METHODS: One-hundred-and-eleven 7-week-old, specific-pathogen-free, male MGs were divided into four groups: normal controls, gerbils with surgically induced DGR, and H. pylori-infected gerbils with and without DGR. Gerbils were killed 4, 12 and 26 weeks after DGR surgery, their stomachs removed and sections prepared. Sections were fixed immediately in 20% phosphate-buffered formalin and subjected to haematoxylin and eosin staining, Alcian blue at pH 2.5/periodic acid-Schiff staining, and immunostaining for smooth muscle cells, H. pylori and 5'-bromo-2'-deoxyuridine (BrdU). RESULTS: The gastric mucosa of H. pylori-infected gerbils showed chronic active gastritis irrespective of DGR throughout the experimental period. The gastric mucosa of H. pylori-infected gerbils with DGR demonstrated higher BrdU labelling than in the other groups. CONCLUSIONS: In MGs, DGR and H. pylori infection synergistically increased gastric mucosal cell proliferative activity. DGR and H. pylori infection may be involved synergistically in gastric carcinogenesis by increasing cell proliferative activity. PMID- 12739709 TI - Stimulated murine macrophages as a bioassay for H. pylori-related inhibition of nitric oxide production. AB - BACKGROUND: Interference with the L-arginine/nitric oxide pathway may be a virulence strategy for the gastric pathogen Helicobacter pylori. This study evaluates a bioassay for such inhibitory actions on nitric oxide synthase. METHODS: Cultured murine macrophages were stimulated by lipopolysaccharide and interferon-gamma. Nitric oxide synthesis and the expression of inducible nitric oxide synthase (iNOS) at increasing concentrations of L-arginine were analysed using chemiluminescence and Western blotting, respectively. RESULTS: The bioassay was evaluated against nitrite accumulation and two established NOS inhibitors. Bacterial extracts or whole cells of one H. pylori strain inhibited nitric oxide production at low L-arginine concentrations (2-20 microM). A higher concentration of L-arginine (200 microM) was not associated with such inhibition. The iNOS expression was not affected by any of the additives compared to stimulated controls. CONCLUSIONS: This bioassay is a reliable and simple method for analysing iNOS inhibition, resolving effects on enzyme activity or enzyme expression. H. pylori water extract and whole cells exert an L-arginine-dependent NOS inhibition, not influencing iNOS expression. PMID- 12739711 TI - Quantitative assessment of motility-associated changes in gastric and duodenal luminal pH in humans. AB - BACKGROUND: Interdigestive pain relieved by food is a common feature of ulcer disease. We tested the hypothesis that the duodenal bulb is intermittently acidified in association with phase III of the interdigestive motility cycle, and tried to quantify the balance between acid and duodenal bicarbonate secretion during this particular period. METHODS: The experiments were performed in Helicobacter-negative healthy volunteers. Gastric and duodenal luminal pH was measured with a triple antimon electrode before, during, and after phase III of the migrating motor complex. Gastric acid secretion rate was measured in real time with a perfusion system and duodenal bicarbonate secretion was estimated from a continuous recording of the transmucosal potential difference (PD) in the duodenal bulb. RESULTS: No significant changes in bulb pH occurred before, during, or after phase III. During the studied time window, the stomach produced 2.24 +/- 0.55 mmol of acid at a peak pH of 1.74 +/- 0.10. Basal HCO3- secretion calculated from bulb PD was 0.82 +/- 0.12 mmol x 30 min(-1) to which was added 0.47 +/- 0.07 mmol of HCO3- during duodenal phase III. The contribution of retroperistalsis-driven HCO3- reflux was small (0.08 +/- 0.02 mmol). CONCLUSIONS: Both the pH recording and the quantitative assessment of secretion rates show that in healthy subjects, fasting gastric acid production and duodenal bicarbonate secretion are of similar magnitude and dynamically coordinated. The mechanism behind the linkage may be reflex activation by motor activity, or a luminal PCO2 rise during phase III activity. PMID- 12739710 TI - Relevance of abdominal gas analysis and transit study after colorectal cancer surgery. AB - BACKGROUND: Colon gas volume analysis using abdominal radiographs is an objective and reproducible method for evaluating functional bowel disorders. The aim of this study was to clarify the relevance of colon gas distribution and transit time in rectosigmoid cancer patients after surgery. METHODS: Segmental colon gas volume score was calculated using plain abdominal radiography and evaluated in 40 patients who had undergone sphincter-saving resection. Segmental colonic transit time was analysed using radiopaque markers in the same patients. RESULTS: Transit times in the right colon (RCT) were 15.3 +/- 1.1 h and in the left colon (LCT) 11.2 +/- 1.1 h. Gas volume scores in the RCS and LCS were 1.10 +/- 0.13% and 1.06 +/- 0.14%, respectively. Neither colonic transit time nor colon gas volume score correlated with the operation methods for rectosigmoid colon cancer. A positive correlation of RCS and a negative correlation of LCS/RCS with ageing were noted in male patients but not in female patients. There was no correlation between RCT and RCS (r = 0.028); however, LCT correlated with LCS (r = 0.318, P < 0.05). The ratio of colonic transit time (LCT/RCT) was 0.84 +/- 0.10, while that of colon gas volume score (LCS/RCS) was 1.29 +/- 0.21. There was a significant correlation between LCT/RCT and LCS/RCS (r = 0.541, P < 0.001). CONCLUSIONS: Analysis of colon gas volume is useful for evaluating colonic transit time in rectosigmoid cancer patients after sphincter-saving surgery. PMID- 12739712 TI - In vivo influence of nicotine on human basal and NSAID-induced gut barrier function. AB - BACKGROUND: Smoking reduces the non-steroidal anti-inflammatory drug (NSAID) induced small intestinal permeability increase in healthy people. It also affects inflammatory bowel disease that is associated with a disturbed gut barrier function. To assess the role of nicotine on barrier function, its influence on basal and NSAID-induced intestinal permeability was studied in healthy volunteers. METHODS: Thirty-one healthy non-smoker subjects performed permeability tests with 51Cr-EDTA and sugar markers (sucrose, lactulose, mannitol, sucralose) before and during 2 weeks of nicotine patch application, and with and without indomethacin intake, respectively. Since smoking has been described as affecting motility, transit measurements were also done with the sodium[13C]-octanoate and lactose-[13C]-ureide breath tests before and during nicotine exposure. Correlations between permeability markers were checked and the influence of gastrointestinal transit was assessed. RESULTS: Nicotine did not affect barrier function in vivo, nor gastric emptying, small-bowel transit time or orocaecal transit. 51Cr-EDTA and lactulose correlated in basal 0-6 h permeability testing (r = 0.529, P < 0.0001), as did 6-24 h excretion of 51Cr EDTA and sucralose (r = 0.474, P < 0.001); 97% and 90% of the subjects had a permeability increase after indomethacin intake for 0-6 h and 6-24 h excretion of Cr-EDTA, respectively. This population proportion is 63% for lactulose/mannitol and 83% for sucralose. CONCLUSIONS: Short-term exposure to nicotine does not alter normal basal or NSAID-induced gut barrier function or transit. 51Cr-EDTA and the respective sugar markers correlate well in in vivo permeability testing in healthy humans. The radioactive test detects more NSAID-induced permeability increase than does the lactulose/mannitol ratio permeability test. PMID- 12739713 TI - Assessment of mucosal inflammation and circulation in response to probiotics in patients operated with ileal pouch anal anastomosis for ulcerative colitis. AB - BACKGROUND: Pouchitis is a common and troublesome condition, and a disturbed microbiological flora and mucosal blood flow in the pouch have been suggested as possible causes. Laser Doppler flowmetry (LDF) has been used successfully to measure gastric and colonic mucosal perfusion in humans. The aim of this study was to evaluate the effect of intervention with probiotics on ileal pouch inflammation and perfusion in the pouch, assessed by endoscopy, histology, fecal calprotectin and LDF. METHODS: A fermented milk product (Cultura; 500 ml) containing live lactobacilli (La-5) and bifidobacteria (Bb-12) was given daily for 4 weeks to 10 patients operated with ileal-pouch-anal anastomosis (IPAA) for ulcerative colitis (UC). Mucosal perfusion was measured with LDF and the degree of inflammation was examined at predefined levels of the distal bowel by endoscopy and histology. Stool samples were cultured for lactobacilli and bifidobacteria and calprotectin were measured before and after intervention. RESULTS: The LDF measurements were reproducible in the pelvic pouch at each of the predefined levels, but did not change after intervention. The mucosal perfusion was reduced in the distal compared to the proximal part of the pouch. Calprotectin levels did not change significantly after intervention. The median endoscopic score for inflammation was significantly reduced by 50% after intervention, whereas the histological score did not change significantly. CONCLUSION: The results suggest that probiotics primarily act superficially, with change of gross appearance of the mucosa at endoscopy, but without significant effect on histological picture, mucosal perfusion or faecal calprotectin, during a relatively short period of 4 weeks. PMID- 12739714 TI - Ascites fluid and plasma calprotectin concentrations in liver disease. AB - BACKGROUND: Calprotectin, a marker of neutrophil activation, has been associated with a poor prognosis in alcohol-induced cirrhosis. The aims were to study concentrations of calprotectin in patients with various liver diseases, and to further investigate the prognostic value of calprotectin in cirrhosis. METHODS: Plasma calprotectin concentrations were determined in 84 patients with alcohol induced liver disease, 32 hepatitis B or C infected patients, 33 patients with liver disease of other aetiologies, 7 patients with combined aetiologies and in 24 patients with malignant disease. Thirty healthy individuals were included as controls. Ascites calprotectin concentrations were determined in patients with ascites (n = 75). Follow-up for survival was performed after a median observation period of 10 months. RESULTS: Increased plasma and ascites calprotectin concentrations were observed in malignant disease compared to non-malignant disease (P < 0.0001). Plasma calprotectin concentrations were low in viral liver disease compared to patients with non-viral liver disease (P = 0.02) and to controls (P = 0.0002). Plasma calprotectin (>median) was a highly significant marker of poor survival in alcohol-induced cirrhosis (P = 0.001), but was of no prognostic value in non-alcohol-induced cirrhosis (P = 0.88). In decompensated cirrhosis high (>upper quartile) ascites calprotectin concentrations were associated with an increased mortality (P = 0.002), as were high (>median) plasma calprotectin levels (P = 0.009). CONCLUSION: The prognostic importance of calprotectin in alcohol-induced cirrhosis is confirmed and demonstrated as specific for alcohol-induced liver disease. Low calprotectin levels are indicated in viral liver disease, and an association between high ascites calprotectin levels and malignant ascites was observed. PMID- 12739715 TI - Reduction of CD45RA isoform expression and decrease in CD4 and CD8 receptor density in lymphocytes of patients with primary biliary cirrhosis. AB - BACKGROUND: The immunological background of primary biliary cirrhosis (PBC) remains largely obscure. METHODS: Using double colour flow cytometry, we estimated the distribution of functionally different lymphocyte subpopulations in the peripheral blood of 25 PBC patients and 18 controls. We examined: 1) the expression of CD3, CD4, CD8, CD19 and CD56 surface receptors, 2) the distribution of lymphocyte subsets bearing 'naive' (CD45RA+) and 'memory' (CD45RO+) phenotypes in both CD4+ and CD8+ cell populations, 3) the expression of an early activation marker (CD69), 4) the distribution of C1.7 mAb binding cytotoxic effectors in CD3+, CD8+ and CD56+ cells. The surface marker expression was evaluated in terms of percentage of positive cells and receptor density. RESULTS: We found: 1) a decrease in the percentage of total CD3+ and CD4+ cells, an unchanged proportion of CD8+ cells but elevated proportion of CD19+ cells and NK lymphocytes; 2) a reduction in the percentage of 'naive' CD4+ but normal proportion of 'naive' CD8+ as well as CD4+ and CD8+ 'memory' cell subsets; 3) a decrease in the density of CD4 and CD8 receptors in the subsets of 'naive' and 'memory' T cells, 4) an increase in the percentage of CD69 receptor bearing T cells but unchanged proportion of C1.7 mAb. CONCLUSIONS: It is concluded that the reduction in number of 'suppressor-inducer-like 'naive' CD4+ T-cell subsets in association with the decrease in fluorescence intensity for CD4 and CD8 may significantly contribute to the mechanisms that could account for a development of PBC. PMID- 12739716 TI - Sampling variability on percutaneous liver biopsy in patients with chronic hepatitis C virus infection. AB - BACKGROUND: Sampling variability on liver biopsy has been demonstrated in a variety of liver diseases. The objective of this study was to determine whether sampling variability exists on percutaneous liver biopsy in patients with chronic HCV infection. METHODS: Two separate tissue samples were obtained from the right lobe of the liver, through a single skin puncture, in 29 patients (22 M, mean age 43.4 +/- 8.1 years) with chronic HCV infection. The biopsies were assessed using a descriptive histological reporting system and Knodell's Histological Activity Index (HAI) and compared for differences in necroinflammatory activity (grade) and fibrosis (stage). RESULTS: Thirteen (44.8%) patients had a difference of > or = 1 grade between the 2 biopsies on the descriptive system and 13 differed by > or = 1 stage. On the HAI, 20 (69.0%) patients had a difference of > or = 2 in the necroinflammatory activity score and 10 (34.5%) had a difference of > or = 4; whereas, 11 (38.0%) patients had a difference of > or = 1 in the fibrosis score and 6 (20.7%) had a difference of > or = 2. The mean difference between the two sets of biopsies was 2.4 +/- 2.1 (range 0-7) for the necroinflammatory activity and 0.6 +/- 0.9 (range 0-3) for fibrosis. Spearman's correlation coefficient (r) was moderate for both necroinflammatory activity (r = 0.53, P < 0.01) and fibrosis (r = 0.62, P < 0.0001). CONCLUSIONS: Sampling variability exists on percutaneous liver biopsy in patients with chronic HCV infection and should be taken into consideration when decisions regarding prognosis and therapy are made based on biopsy, and when defining histological response to antiviral regimens. PMID- 12739717 TI - A simplified method for computed tomographic estimation of prognosis in acute pancreatitis. AB - BACKGROUND: Computed tomography is valuable for the diagnosis of acute pancreatitis. Although CT-based prognostic scoring systems are available, they are complex and impractical for routine clinical use. We examined the validity of a simplified CT-based scoring system in a cohort of patients with acute pancreatitis. METHOD: Observational study based on correlation of CT findings with clinical outcomes. Seventy patients admitted to the Royal Infirmary of Edinburgh from January 1991 to December 1997 with a diagnosis of acute pancreatitis and undergoing CT with intravenous contrast during the first 3 to 10 days after admission were included in the study. RESULTS: Multivariate logistic regression analysis demonstrated that the finding of mesenteric oedema and free peritoneal fluid on CT were independent early predictive factors of adverse outcome. Allocating one point each for either mesenteric oedema (MO) or peritoneal fluid (P) (giving a maximum score of 2), a simple MOP score was derived. Compared with the Glasgow and APACHE multiple-factor scoring systems and the Helsinki and Balthazar CT-based scoring systems areas under ROC curves were: admission Apache II 0.57, admission Glasgow 0.62. Balthazar score 0.79, Helsinki score 0.85 and MOP score 0.87. CONCLUSIONS: The presence of mesenteric oedema or peritoneal fluid on CT appears to be a simple and widely applicable predictor of disease severity in acute pancreatitis. PMID- 12739718 TI - Expression of mucosal addressin cell adhesion molecule-1 (MAdCAM-1) during small bowel graft rejection in rats. AB - BACKGROUND: Mucosal addressin cell adhesion molecule-1 (MAdCAM-1) is a critical endothelial adhesion molecule for lymphocyte trafficking to gut-associated lymphoid tissues (GALT) under physiological conditions. It is expressed on special postcapillary venules, that is the high endothelial venules (HEV) in GALT. In this study, we investigated changes in MAdCAM-1 expression during small bowel graft rejection. METHODS: Orthotopic small-bowel transplantation (SBT) with portocaval drainage was performed from brown Norway (BN) rats to Lewis (LEW) rats. Isografted (LEW --> LEW) and untransplanted animals served as controls. Animals were killed on days 3, 4, 5, 6 and 7 after SBT. Cryostat sections (6 microm thick) were prepared from normal small-bowel tissues and small-bowel grafts, including Peyer patches (PPs). Indirect immunoperoxidase staining was performed using monoclonal antibodies (mAbs) against rat MAdCAM-1 (OST12). RESULTS: In the PPs of controls, MAdCAM-1 antibodies specifically stained the endothelial cells of HEV, which were predominantly located in the interfollicular areas. In the allografts on day 4 after SBT, the MAdCAM-1 expression was weaker on the HEV in the PPs than in the controls. In the lamina propria of controls, a faint expression of MAdCAM-1 on vessels was observed. On day 4 after allogeneic SBT, the MAdCAM-1 was more strongly expressed on the endothelial cells of the vessels at the base of the villi. As rejection developed, the MAdCAM-1 expression on the vessels progressed toward the villus tip. CONCLUSIONS: The change of MAdCAM-1 expression may be involved in the development of small-bowel graft rejection. The vessels at the base of villi, which is associated with lymphocyte recruitment, may become a site of intense immune reactivity in the early phase of small-bowel allograft rejection. PMID- 12739719 TI - Intravascular ultrasonography: a promising imaging tool in transjugular intrahepatic portosystemic stent shunts. PMID- 12739720 TI - Macroamylasaemia, IgA hypergammaglobulinaemia and autoimmunity in a patient with Down syndrome and coeliac disease. AB - The association between macroamylasaemia and coeliac disease in Down syndrome with multiple autoimmune abnormalities has never been reported. A 40-year-old woman with a 15-year history of immunoglobulin A and immunoglobulin M hypergammaglobulinaemia, chronic diarrhoea, persistent mild aspartate aminotransferase (AST) elevation and anaemic syndrome was admitted to hospital because in the previous 3 months she had developed amenorrhoea, dizziness, alopecia, constipation, pallor and asthenia. Biochemical and immunological analyses showed macroamylasaemia. The patient presented clinical and intestinal histopathological features of coeliac disease. Immunological abnormalities included the presence of antigliadin, antiendomysium, antitransglutaminase, antinuclear, antismooth muscle and anti-SSA/Ro antibodies. Macroamylase resulted in a complex of amylase and immunoglobulin A. Later clinical follow-up of a gluten-free diet showed a transitory decrease in seric immunoglobulin A and macroamylase with persistent autoantibodies and AST elevation. An intestinal mucosal immune disorder could lead to coeliac disease and macroamylasaemia in a patient with Down syndrome presenting other immune alterations. PMID- 12739721 TI - Management of gastric submucosal polyps by UK specialists. PMID- 12739722 TI - Global solutions to waste globalisation. PMID- 12739723 TI - The use of waste materials in asphalt concrete mixtures. AB - The purpose of this study was to investigate (a) the effects of rubber and plastic concentrations and rubber particle sizes on properties of asphalt cement, (b) on properties of asphalt concrete specimens and (c) the effects of fly ash, marble powder, rubber powder and petroleum contaminated soil as filler materials instead of stone powder in the asphalt concrete specimens. One type of limestone aggregate and one penetration-graded asphalt cement (75-100) were used. Three concentrations of rubber and plastic (i.e. 5%, 10% and 20% of the total weight of asphalt cement), three rubber particle sizes (i.e. No. 4 [4.75mm] - 20 [0.85 mm], No. 20 [0.85mm] - 200 [0.075mm] and No. 4 [4.75mm] - 200 [0.075mm]) and one plastic particle size (i.e. No. 4 [4.75mm] - 10 [2.00mm]) were also used. It was found that while the addition of plastic significantly increased the strength of specimens, the addition of rubber decreased it. No. 4 [4.75mm] - 200 [0.075mm] rubber particles showed the best results with respect to the indirect tensile test. The Marshall stability and indirect tensile strength properties of plastic modified specimens increased. Marble powder and fly ash could be used as filler materials instead of stone powder in the asphalt concrete pavement specimens. PMID- 12739724 TI - Data reconciliation, structure analysis and simulation of waste flows: case study Vienna. AB - The management of complex waste flow systems requires a systematic approach for the handling of data, for obtaining a consistent picture of the system under consideration, and for simulating various policy scenarios and evaluating material control strategies. In this paper the implementation of a useful methodology is presented, which has been developed in previous works and is further enhanced for modelling, identifying, analysing and simulating material flow systems for which at most one measurement per flow is available for a single balancing period. The methodology enables the analyst to cope with missing data and uncertainty in the measurements. A data reconciliation procedure is used to minimise the uncertainty concerning flows by exploiting the redundancies created by restricting the available data to fulfil the available structural information. Statistical tests are introduced to enable the user to check the compatibility of the data with the a priori information. The origins analysis and destination analysis tools allow for a deeper insight into the system structure. Policy scenarios can be treated using the simulation tools. The waste flow system of the city of Vienna has been chosen to demonstrate step-by-step the procedure for building a reliable model and the effective application of the above mentioned methods and tools. Current and future research focuses on models balancing different interrelated quantities simultaneously and on incorporating stock accumulation and depletion behaviour. PMID- 12739725 TI - Slag from hazardous waste incineration: reduction of heavy metal leaching. AB - Hazardous waste incineration (HWI) in rotary kilns and the disposal of the residues on landfills play an important role in German waste treatment. In order to reduce costs by disposal on cheaper landfill sites still applying to landfill regulations the leaching behaviour of HWI-slag should be improved further. In a new process-integrated approach hazardous waste is mixed with limestone, which initiates chemical reactions with heavy metals in the rotary kiln yielding new compounds of different solubility. These reactions were observed after treatment at 1200 degrees C combined with fusion processes, at 930 degrees C they also occurred without fusion to the major part. For that purpose HWI-slag/limestone mixtures are thermally treated and then examined by elution tests. A minimum of overall heavy metal leaching was determined at CaO-contents between 15 and 20% after sintering at the average temperature at HWI. PMID- 12739726 TI - Modelling simplification of landfill processes by using methods of reliability theory. AB - By the year 2005 most landfills in Germany will be closed down and begin the closing phase and later the after-care phase. So far there have been only a few scientific investigations concerning the hazard potential of stored waste for the environment. For this reason, it is necessary to predict how the hazard potential of landfills will vary after closure. The prediction can be made by describing the coupled processes of biodegradation and pollutant transport using models that are currently being developed. These models deal with highly non-linear differential equations having a large number of parameters. In order to permit application of the model in practice, it is necessary to reduce these to parameters which are essential to the reliability of the landfill. They have to be identified by sensitivity investigations with the aid of methods of reliability theory so that models can be limited to these essential parameters and the calculation effort as well as measurement of parameters (monitoring) can be considerably reduced. PMID- 12739727 TI - Serial water balance method for predicting leachate generation in landfills. AB - This paper presents the "Serial Water Balance" method for predicting leachate generation in landfills. This procedure makes it possible to calculate the total leachate likely to be generated, by estimating an individual cell by cell water balance. This new development considers the interaction effects between cells, through the execution of simple field capacity tests on solid waste samples under different loading conditions. The procedure described in this paper simulates the effect induced by fluid percolating from a cell at an upper layer to cells in the immediately subjacent layer. The method described here was applied to the landfill in Nuevo Laredo, Tamaulipas (Mexico), after first ascertaining information regarding: duration of construction time, exposure time, surface area, the quantity of waste, number of confining cells, and local weather conditions. In a full-scale test case the suggested method has predicted 67% of the leachate produced in a period of 4 years. Further testing and more detailed analysis of the composition of the waste used in the calculations, may improve the accuracy in predicting leachate production. Even though this SWB methodology was applied to a landfill subject to extreme climatological conditions (high daytime temperatures), it is possible to adapt the methodology to solid waste disposal sites in regions with more humid or moderate climates. PMID- 12739728 TI - Determination of field capacity of municipal solid waste with surcharge simulation. AB - The following study was carried out as part of the environmental monitoring of a landfill in Nuevo Laredo, Tamaulipas (Mexico). The parameter of field capacity is important in predicting the amount of leachate generated by solid waste disposal sites, because of the polluting potential of leachate. This paper describes how the field capacity for municipal solid waste was determined, and the purpose of this document is to present a methodology, and to describe the devices designed for determining the field capacity of municipal solid waste. The method consists of applying a surcharge to a representative sample of rubbish, to simulate the effects of the overlying layers in a landfill. The experimental results showed that the higher the compaction of the sample, the smaller the amount of water required to satisfy the field capacity and thus to start the leaching process. Standardisation of the methodology for determining this parameter is required in order to be able to compare the results with those obtained in other parts of the world. PMID- 12739729 TI - Kinetics of anaerobic treatment of landfill leachates combined with urban wastewaters. AB - The anaerobic degradation of landfill leachates mixed with domestic wastewater has been studied in a pilot-scale Upflow Anaerobic Sludge Blanket (UASB) reactor. A previous work in laboratory-scale had shown that a fraction (5%) of the refractory organic matter could be additionally degraded when these two substrates were treated in conjunction, but this synergistic effect in the Chemical Oxygen Demand (COD) removal was not reproduced. However, the mass loading rate for which the maximum degradation was obtained was higher for the mixtures (0.5 kg COD/kg SSV x d) than for the separated components (0.18 and 0.19), allowing an increase in the treatment capacity of the leachates. The methane productivity (304 L/kg COD) was close to the theoretical maximum and independent of the proportion of the mixture components. The experimental data were fitted to a modification of Haldane's kinetic model, in which the parameters depend on the hydraulic residence time and the biomass concentration. PMID- 12739730 TI - Magnetic susceptibility variation of MSW compost-amended soils: in-situ method for monitoring heavy metal contamination. AB - Magnetic susceptibility was measured for agricultural soils in Mornag area, Tunisia, where the soils were partly amended by manure or compost obtained from municipal solid waste stabilisation ('MSW compost'). Our study indicates that natural non-treated soils and manure-amended soils are always low in magnetic susceptibility, but MSW compost-amended soils show higher values of this parameter. Actually, the increase of magnetic susceptibility shows a direct correspondence with the increasing of the amount of MSW compost added to the soil. According to the magnetic mineralogical investigation carried out by isothermal remanent magnetisation acquisition technique, higher magnetic susceptibility values are depending on an increase in ferromagnetic components such as either magnetite (beta-Fe3O4) or maghemite (gamma-Fe2O3) particles. The growth in content of these ferromagnetic components corresponds to an increase of the concentration of heavy metals in soils, which means that magnetic susceptibility indirectly indicates the concentration of heavy metals in MSW compost-amended soils. PMID- 12739731 TI - Carbon mineralisation and plant growth in soil amended with compost samples at different degrees of maturity. AB - The carbon and nitrogen mineralisation of a composting mixture of brewing yeast and lemon tree prunings was studied, at different degrees of stabilisation of this matrix, within an incubation experiment in soil. Meanwhile, a growth test in pots with ryegrass (Lolium perenne L.) was carried out using the selected soil and equal amounts of the composting mixture taken at different maturation steps, in order to evaluate the additions of these organic amendments in terms of fertilising value. Samples of the composting mixture, when poorly transformed through the biostabilisation process, showed high CO2-C releases in the soil, due to the microbial attack on easily degradable organic fractions still present in the mixture, with 24.7% mineralisation of the initial total organic carbon (TOC) after a 70 day incubation. On the other hand, mature compost was the most stable matrix, with only 5.4% of TOC mineralised after 70 days. Furthermore, amendments with the initial composting mixture led to negative net N-mineralisation during 56 days of incubation with soil. Only slight negative values of the net N mineralisation were detected with fully stabilised compost. Nevertheless, pot experiments with ryegrass revealed that mature compost may promote N mineralisation to certain extents. Moreover, mature compost did not produce any phytotoxic effect, behaving as a slow-action organic fertiliser with N made available through a progressive mineralisation. Thus, the results gained through this study are a confirmation that the fertilising quality of a compost destined for agricultural uses is heavily affected by the complete exhaustion of the maturation reactions. PMID- 12739732 TI - Don't ban PVC: incinerate and recycle it instead! AB - Plastics are making a growing contribution to sustainable development. For example, over an expected lifetime of 50 years, the use of window frames and insulating materials made of plastic in buildings save many times the energy required to manufacture them. Plastics for packaging purposes provide protection against damage and dirt contamination, thereby saving considerable amounts of material and energy. Choosing appropriate disposal strategies for plastic waste also helps to protect the environment (Mark 2000). PMID- 12739733 TI - Acromegaly: disease or syndrome? Not only a semantic question. PMID- 12739734 TI - Acromegaly: are new tests needed? PMID- 12739735 TI - Relapses of hyperthyroidism in patients treated with radioiodine for nodular toxic goiter: relation to thyroid autoimmunity. AB - Relapses of hyperthyroidism after treatment with radioiodine for uni- or multi nodular goiter may be accompanied by the appearance of TSAb. However, this phenomenon has only emerged from one retrospective study on Northern European patients, in which it was not possible to determine whether TSAb also appeared in treated patients who did not relapse. The present study aimed to assess the appearance, immunogenic nature and clinical characteristics of hyperthyroidism relapse after treatment with 131I for nodular toxic goiter in patients from the Mediterranean area. A retrospective study was performed on 76 consecutive patients, born and resident in Sicily and aged 56-80 yr at diagnosis, who were treated with radioiodine. Serum aliquots obtained from the patients before 131I treatment and during a follow-up of 36-144 months were assayed for TSAb and TPOAb. The clinical charts of the patients were also re-examined. Twenty-six out of 76 patients (36.8%) had a hyperthyroidism relapse after a first treatment with 131I. Eight of the 26 (30.7%) also relapsed after the second treatment. Three out of 26 (11.5%) relapsed after a third treatment. The 50 patients who required only one treatment and the 18 who relapsed only once were all TSAb-negative at baseline, while 3/8 (37.5%) who relapsed also after the second treatment were already TSAb-positive at baseline. TSAb became positive in 3/18 patients (16.7%) who relapsed once, and in 4/8 (50.0%) of those who relapsed after a second treatment. One of these 7 TSAb-positive relapsers was also already TPOAb-positive at baseline and another became TPOAb-positive after treatment. The presence of circulating TSAb in 3/76 (3.9%) patients before treatment for toxic goiter more probably points to a diagnosis of Marine-Lenhart's syndrome. In contrast, the de novo appearance of TSAb in the presence of hyperthyroidism relapse in 4/76 (5.3%) patients suggests the development of a Graves'-like disease after radioiodine treatment. This occurrence does not seem to have precise ethnic grounds, since the incidence we observed in Mediterranean patients was similar to that previously reported in Northern European patients. PMID- 12739736 TI - Effect of small doses of dexamethasone on plasma leptin levels in normal and obese subjects: a dose-response study. AB - To further elucidate the role of glucocorticoids in the regulation of leptin secretion, we studied the effects of overnight small doses of dexamethasone on plasma leptin levels in normal weight controls and in obese patients and correlated the results with indexes of insulin sensitivity and body fat distribution. In 114 subjects (81 obese patients, 49 women and 32 men, BMI 37.4 +/- 0.77 kg/m2 and 33 normal-weight subjects, 17 women and 16 men, BMI 22.1 +/- 0.41 kg/m2) plasma F and leptin levels were measured at 08:00 h basally and after the administration of different doses of dexamethasone (a fixed dose of 1-mg and 0.0035, 0.007, 0.015-mg/kg bw, given po at 23:00 h the night before). Tests were performed one week apart with bw remaining stable over the study period. Basal leptin levels were significantly higher in obese than in normal subjects (31.9 +/ 2.41 vs 7.7 +/- 0.93 ng/ml, p<0.0001). In obese patients, leptin levels increased significantly by 1-mg (from 31.9 +/- 2.41 to 35.0 +/- 2.59 ng/ml, p<0.005) and the 0.015-mg/kg bw dose (from 31.5 +/- 2.34 to 33.7 +/- 2.44 ng/ml, p<0.05), while they were unaffected by each dose of dexamethasone in normal subjects. However, after splitting subjects by gender, mean leptin levels rose from 39.3 +/- 2.97 to 43.3 +/- 3.12 ng/ml after the 1-mg dose, p<0.005, from 39.1 +/- 2.87 to 43.6 +/- 2.91 ng/ml after the 0.015-mg/kg bw dose, p<0.005, from 39.3 +/- 2.90 to 42.2 +/- 2.90 ng/ml after the 0.007-mg/kg bw dose, p<0.05 and from 38.8 +/- 2.66 to 41.1 +/- 2.87 ng/ml after the 0.0035-mg/kg bw dose, p=0.055, only in obese women. Conversely, no leptin changes were seen in the other groups and no differences were observed in the leptin response between groups. After the 1-mg dose, in the whole group, the absolute leptin variation was weakly but significantly related to BMI values (r=0.231, p<0.02) while in all sessions the percent leptin changes over baseline were not significantly correlated with age, BMI, waist, WHR, insulin, HOMA index, a marker of insulin sensitivity, plasma dexamethasone concentrations and to the percent cortisol variation following dexamethasone. In conclusion, in obese women but not in obese men and in normal weight subjects, small overnight increases in plasma glucocorticoid concentrations induced gender-related plasma leptin elevations that were unrelated to body fat distribution and insulin sensitivity. A greater sensitivity of female adipose tissue to glucocorticoids probably underlies this sexually dimorphic pattern of leptin response. These findings provide an additional piece of information on the regulation of leptin secretion exerted by glucocorticoids. PMID- 12739737 TI - GH release after GHRH plus arginine administration in obese and overweight women with polycystic ovary syndrome. AB - Few and unclear data are available in the literature about the relationship between impairment of GH/IGF-I axis and polycystic ovary syndrome (PCOS). This study was aimed to evaluate the basal GH and IGF- levels, and GH release after challenge test in obese and overweight women with PCOS. Thirty patients with PCOS and other 30 healthy women matched for age, body mass index (BMI) and waist-hip ratio (WHR) were studied. Serum follicle-stimulating hormone (FSH), LH, PRL, E2, P, 17OH-progesterone (17OH-P), total T, delta4, DHEA-S, SHBG, GH and IGF-I levels were evaluated in each subject. A GHRH plus arginine challenge test was performed in all subjects. After provocative test, in PCOS and control women the GH levels were significantly (p<0.05) higher in comparison to basal values from 30 min to 120 min. At the same times, a significant (p<0.05) difference was observed between women with PCOS in comparison to healthy women. The mean peak value of GH resulted significantly (p<0.05) lower in PCOS women in comparison to healthy women. The total GH response (area under curve, AUC) to GHRH plus arginine test resulted significantly (p<0.05) lower in PCOS than in healthy women. These findings were statistically significant (p<0.05) also considering the distinction in obese and overweight women. The AUC for GH secretion was significantly lower (p<0.05) in obese in comparison to overweight subjects in the control group, whereas no significant difference was detected between obese and overweight women in the PCOS group. In conclusion, in PCOS women there is a BMI-independent alteration of the GH levels. Further investigations will be necessary to establish the real cause of these data. PMID- 12739738 TI - Three-hour spontaneous GH secretion profile is as reliable as oral glucose tolerance test for the diagnosis of acromegaly. AB - The diagnosis of acromegaly, in an appropriate clinical context, usually relies on lack of GH suppression below 1 microg/l during OGTT coupled with elevated IGF I levels. On the other hand, in normal subjects glucose-induced inhibition of GH secretory bursts without any further decrease of interpulse GH levels had already been shown. Based on the foregoing, we aimed to compare the diagnostic reliability of OGTT-induced GH nadir with that recorded during 3-h spontaneous GH secretion. In 59 acromegalic patients (17 male and 42 female, age, mean +/- SE 51.5 +/- 1.9, range 21-76 yr) and in 82 normal subjects (43 male and 39 female, age, mean +/- SE 35.7 +/- 1.5, range 15-72 yr) GH secretion was evaluated every 30 min from 0 to 180 min during slow saline infusion or OGTT (75 g at 0 min). A nadir GH concentration below 1 microg/l was recorded in all normal subjects either during OGTT or saline infusion if GH secretion was evaluated over 180 min. In contrast in acromegalic patients a nadir GH concentration below 1 microg/l never occurred in both conditions. This study shows that a 3-h spontaneous GH profile is as reliable as OGTT in the diagnosis of active acromegaly. PMID- 12739739 TI - Is it useful to routinely biopsy hot nodules in iodine deficient areas? AB - Incidence of nodular thyroid disease as well as that of functioning thyroid nodules (FTN) increases dramatically in iodine deficient (ID) areas. Cancer is extremely rare in FTN; thus, some do not routinely biopsy and treat them with radioactive iodine (RAI) straight away or follow-up. The outcome of 296 patients followed or treated at our institution for solitary or multiple FTN were retrospectively evaluated. Hospital records of 224 female, 72 male patients, with a mean +/- SD age of 54.9 +/- 12.4 yr and followed for 22 (0-156) months were examined. 175 patients had solitary, 121 had multiple hot or warm nodules. 230 (77.7%) of the patients received RAI treatment. 402 fine needle aspiration biopsies (FNABs) were performed on 260 patients and on 343 FTN (381 benign and 21 suspicious diagnoses). Eleven of the patients were operated for suspicious FNAB results and 10 were followed-up. Only one nodule turned out to be malignant. Malignancy is extremely rare in functioning thyroid nodules (0.34%) and some of malignant cases could be predicted by their suspicious clinical features. Routine practice of treating FTN with RAI therapy is reasonable in clinically low-risk patients. FNAB is reserved for cases with suspicious clinical features, resulting in fewer surgeries and reduced cost. PMID- 12739740 TI - Identification of genes in thyrocytes regulated by unfolded protein response by using disulfide bond reducing agent of dithiothreitol. AB - Disulfide bonds are formed between the sulfhydryl groups in two cysteine residues of a protein. The formation of these bonds is necessary for the proper folding of a protein into its active three-dimensional form. In this study, the genes associated with disulfide bond formation of proteins from the rat thyroid cell line, FRTL-5 cell, were investigated using disulfide bond reducing agent of dithiothreitol (DTT), which prevented disulfide formation of newly synthesized proteins. The expression of six genes, they being the cAMP phosphodiesterase 7A1, neuronal cell death inducible putative kinase (NIPK), cytosolic LIM protein (Ajuba), Eker, early growth response 1 and the ferritin heavy chain, was specifically enhanced under both reductive conditions and various endoplasmic reticulum (ER) stresses inducing drugs such as Brefeldin A (BFA), calcium ionophore A23187 (A23187) and tunicamycin. These results suggest that a suitable redox environment is necessary for the correct disulfide bond conformation in thyrocytes in a complex system. PMID- 12739741 TI - Changes of autonomic cardiac profile after a 3-week integrated body weight reduction program in severely obese patients. AB - The autonomic control of the heart is abnormal in obese subjects due to a prevalence of sympathetic over parasympathetic limb of the autonomic balance. We evaluated the effects of a short-term (3 weeks) integrated body weight reduction program (consisting of energy restricted diet and high-intensity exercise training) on heart rate variability (HRV) in severely obese, normotensive patients. The HRV was evaluated both in the time and frequency domain over a 18 hour Holter recording period obtained before and at the end of the third week. Three-week body weight reduction program reduced BMI (from 41.4 +/- 4.6 to 39.5 +/- 4.3 kg/m2, -4.6%, p<0.0001) and heart rate (from 77.8 +/- 8.6 to 73.6 +/- 8.7 b/min, p=0.0003). Significant changes in the autonomic profile were observed both in the time and frequency domain (SD of RR interval, SDRR: +16.1%; mean squared successive difference: (MSSD) +16.7%; percentage of RR intervals differing more than 50 msec from the preceding one, pNN50: +31.8%; low frequency oscillation, LF: +17.1%; high frequency oscillation, HF: +/- 18.2%). In conclusion, this study demonstrates that a short-term, integrated body weight reduction program is able to favorably modify the autonomic profile in a population of normotensive, severely obese subjects. The reduction of heart rate and the increase in parasympathetic activity may consistently contribute to a reduction of the risk of cardiovascular morbidity and of sudden cardiac death, still high in this patients' group. PMID- 12739742 TI - GH-releasing peptide (GHRP-6)-induced ACTH release in patients with addison's disease: effect of glucocorticoid withdrawal. AB - GH releasing peptide (GHRP-6) is a synthetic hexapeptide with potent GH releasing activity both in man and in animals. This peptide is also able to stimulate ACTH and cortisol (F) release. It has been suggested that the ACTH responsiveness to GHRP-6 is modulated by circulating glucocorticoid levels. To further clarify this hypothesis, we studied the effect of GHRP-6 (1 ug/kg, iv) on ACTH and F release in patients with Addison's disease (no.=6) during replacement therapy and after 72 h of glucocorticoid withdrawal. Seven controls were also submitted to a single GHRP-6 test. In control subjects, ACTH values (pmol/l; mean +/- SE) increased from 2.9 +/- 0.8 to 4.7 +/- 1.4 (peak). AUC (pmol.min/l) values were 170.3 +/- 48.8. F (nmol/l) values increased from 257.0 +/- 42.9 to 367.0 +/- 50.8. In patients with Addison's disease there was an increase in ACTH levels from 38.1 +/ 17.1 to 174.9 +/- 79.4 after GHRP-6 administration. AUC values were 5490.4 +/- 2269.1. After 72 h withdrawal of glucocorticoid, there was an increase in basal ACTH values (191.2 +/- 97.3), and a trend toward an increase in ACTH levels after GHRP-6 (p=0.053). Patients with Addison's disease on therapy showed a significantly higher ACTH response to GHRP-6 when compared to controls. Our results show that in patients with Addison's disease on replacement there is an increased ACTH release after GHRP-6 administration, compared to controls. After 72 h glucocorticoid withdrawal, this enhanced responsiveness is not maintained. Our data suggest that circulating glucocorticoids modulate GHRP-6-induced ACTH release and that multiple mechanisms may be involved in this process. PMID- 12739744 TI - Transition from pediatric to adult care of children with chronic endocrine diseases: a survey on the current modalities in Italy. AB - We report on a survey carried out in 65 pediatric and adult endocrinological centers concerning: 1) the modalities of the transfer of children with chronic endocrine diseases from pediatricians to adult endocrinologists, 2) opinions and suggestions from physicians of the Centers, and 3) specific details regarding GH deficiency. The main results are: 1) The mean age of transfer is around 18 yr of age. 2) The reasons for the transfer are personal convincement of pediatricians in 47%, administrative reasons in 37% and patient's desire in 16% of cases 3) In the majority of cases a discharge summary is sent by the pediatrician to the endocrinologist often followed by a phone call, whereas 30% of endocrinologists do not send a report back to pediatricians. 4) Less than half of the Centers are satisfied with the modalities of the transfer and the remainder complain about the lack of communication, no common guidelines, and differences in the management of patients. However, all are willing to try to improve this important time for adolescents with chronic diseases. 5) As far as GH deficiency is concerned, the main differences between pediatric and adult endocrinological centers are the different tests used to re-evaluate the diagnosis and the higher doses of GH used by pediatricians to treat young adults. In conclusion, considering the interest and desire of physicians, a structural intervention of the scientific societies to help to overcome problems is highly desirable. PMID- 12739743 TI - Macroprolactinemia: predictability on clinical basis and detection by PEG precipitation with two different immunometric methods. AB - Prolactin (PRL) in human serum is present in three species: monomeric PRL (23 kDA), big PRL (50-60 kDa) and big, big PRL (bb-PRL or macroprolactinemia) of 150 170 kDa. Macroprolactin seems to be mainly composed of a molecule of monomeric PRL and an immunoglobulin G anti PRL. Its biological activity is considered low or absent, but it is measured, at various degrees, by the immunoassay method, thus causing diagnostic problems. Polyethylene Glycol (PEG) has been employed to precipitate macroprolactin, allowing its detection. This method is not applicable to all immunoassays for technical reasons. Our aim was to evaluate: 1) the predictability of macroprolactin on a clinical basis; 2) the possibility of applying PEG precipitation to Abbott AxSYM analyzer beside Roche Elecsys (already approved). We classified 34 hyperprolactinemic women, on a clinical and imaging basis, in four groups: A: functional hyperprolactinemia; B: pituitary lesions hyperprolactinemia; C: probably macroprolactinemia; D: unclassifiable hyperprolactinemia and a "control" group E of 19 healthy women. PRL was assayed, both with Elecsys and AxSYM, before and after PEG serum treatment. Eleven out of twelve group C, and 5/7 group D patients showed macroprolactinemia, against 1/7 in A and 1/8 in B. PEG was suitable for AxSYM only after the same treatment of the calibration standards, thus performing outcomes overlapping Elecsys. For clinical purposes, in the presence of macroprolactinemia, besides the recovery ratio, molar or ponderal monomeric PRL assay should be calculated. PMID- 12739746 TI - Management of post-menopausal osteoporosis: something new on the horizon? PMID- 12739745 TI - Acromegaly due to ectopic secretion of GHRH by bronchial carcinoid in a patient with empty sella. AB - GH hyperproduction due to ectopic secretion of GHRH is a rare cause of acromegaly. Since 1959, approximately 50 cases of ectopic GHRH production from extrapituitary tumors have been described. Here we report the clinical and biochemical features of a 47-yr-old Caucasian woman with ectopic GHRH syndrome sustained by a bronchial carcinoid. The criteria for the diagnosis of acromegaly due to ectopic GHRH secretion were satisfied in our patient (i.e. confirmation of active GH hypersecretion, unequivocal demonstration of GHRH production and secretion from an extrapituitary tumor and cure of acromegaly after neoplasm removal). The tumor was an atypical carcinoid and there was a familial history of lung and colorectal cancer. Acromegaly was slightly active (mean GH value: 7.4 ng/ml, IGF-I: 436 ng/ml) and after tumor removal there was a progressive decline of GH levels, consistent with remission of pituitary somatotroph hyperplasia. Pituitary radiology showed an empty sella demonstrating for the first time its association with ectopic GHRH syndrome. PMID- 12739747 TI - Antithyroid drug treatment prior to radioiodine therapy for Graves' disease: yes or no? PMID- 12739748 TI - Role for interferon-gamma inducible chemokines in endocrine autoimmunity: an expanding field. AB - The chemoattractant cytokines (chemokines) have been classified into 4 major sub families in relation to the position of the cysteine residues in their NH2 terminal portion. Interferon-gamma inducible chemokines (CXCL9/Mig, CXCL10/IP-10, CXCL11/I-TAC), strongly associated to Th1-mediated immune responses, belong to the CXC sub-family. They represent an exception among chemokines in that they specifically interact with a single type of receptor, named CXCR3. A statistically significant increase of CXCL10/IP-10 and CXCL9/Mig expression, in thyroid tissue specimens obtained from subjects affected by Hashimoto's thyroiditis and recent onset Graves' disease has been reported. Furthermore, a statistically significant increase in serum CXCL10/IP-10 levels has been found in newly diagnosed Graves' patients when compared to healthy subjects as well as patients with long standing disease and a strong statistically significant inverse correlation between circulating CXCL10/IP-10 levels and disease duration has been demonstrated. Similar findings have been obtained when Type 1 autoimmune diabetes affected patients have been taken into account. In conclusion, such experiences have demonstrated an important role played by interferon-gamma inducible CXC chemokines in the pathogenesis of glandular autoimmunity. In fact, it is reasonable to assume that glandular epithelial cells may modulate the autoimmune process at least in its initial phase, through the production of chemokines which induce migration of Th1 lymphocytes into the gland. Interferon gamma secretion by lymphocytes would, in turn, stimulate chemokines production by follicular cells, thus perpetuating the autoimmune cascade. PMID- 12739749 TI - Testosterone and coronary vascular tone: implications in coronary artery disease. AB - The greater incidence of coronary artery disease in men compared to women has often suggested possible harmful effects of male sex steroids that could promote coronary atherogenesis and vasoconstriction. However, antiatherogenic and coronary vasodilator effects of testosterone have also been suggested. The interaction of testosterone (T) with its specific receptors may trigger not only long-term genomic effects, but also acute non-genomic vasodilator responses. Testosterone may activate the endothelium and stimulate the nitric oxide-cGMP and/or the hyperpolarization-mediated vascular relaxation pathway. T may also inhibit the signaling mechanisms of smooth muscle contraction such as [Ca2+]i and protein kinases. The T-induced stimulation of endothelium-dependent mechanisms of vascular relaxation and inhibition of the mechanisms of coronary smooth muscle contraction represent potential beneficial effects of T against coronary artery disease. PMID- 12739750 TI - Neuroengineering modeling of single neuron and neural interface. AB - The single neuron has attracted widespread attention as an elementary unit for understanding the electrophysiological mechanisms of nervous systems and for exploring the functions of biological neural networks. Over the past decades, much modeling work on neural interface has been presented in support of experimental findings in neural engineering. This article reviews the recent research results on modeling electrical activities of the single neuron, electrical synapse, neuromuscular junction, and neural interfaces at cochlea. Single neuron models vary form to illustrate how neurons fire and what the firing patterns mean. Focusing on these two questions, recent modeling work on single neurons is discussed. The modeling of neural receptors at inner and outer hair cells is examined to explain the transforming procedure from sounds to electrical signals. The low-pass characteristics of electrical synapse and neuromuscular junction are also discussed in an attempt to understand the mechanism of electrical transmission across the interfaces. PMID- 12739751 TI - Uncovering the mechanisms of deep brain stimulation for Parkinson's disease through functional imaging, neural recording, and neural modeling. AB - High-frequency deep brain stimulation (DBS) of the thalamus or basal ganglia represents an effective clinical technique for the treatment of several medically refractory movement disorders, including Parkinson's disease. However, understanding of the mechanisms of action of DBS remains elusive. The goal of this review is to address our understanding of the effects of high-frequency stimulation within the central nervous system based on results from functional imaging, neural recording, and neural modeling experiments. Using these results, we address the main hypotheses on the mechanisms of action of DBS and conclude that stimulation-induced desynchronization of network oscillations represents the hypothesis that best explains the presently available data. PMID- 12739752 TI - High-resolution spatio-temporal functional neuroimaging of brain activity. AB - The past decades have shown extraordinary progress in our ability to noninvasively image the functions of the human brain. Of particular interest is the recent trend in combining information from electrophysiological and magnetic resonance imaging, which we termed eMRI, to achieve high-resolution functional neuroimaging in both space and time domains. In this article, we review the recent progress in high-resolution functional neuroimaging, in particular the multimodal integration of electroencephalography (EEG) and magnetic resonance imaging (MRI). The state-of-the-art EEG inverse solutions based on different brain electric source models and various approaches to integrate the information from MRI are reviewed. The remaining challenges, future trends, and potential applications of the high-resolution functional neuroimaging research are discussed. PMID- 12739753 TI - Biomechanics of single chondrocytes and osteoarthritis. AB - Osteoarthritis is a significant, debilitating disease that afflicts millions of Americans, yet its etiology is poorly understood. However, there is substantial evidence that biomechanical factors play a role in the development and progression of osteoarthritis. Previous work has demonstrated that biomechanical factors such as an acute insult or the cumulative effects of repetitive loads can induce degenerative changes in joints, cartilage explants, and isolated chondrocytes. Nevertheless, all of these studies suffer from the limitation that the precise nature of the mechanical loads experienced by individual cells is not well defined. Implementation of a single-cell approach, employing existing cell mechanics methodologies and molecular techniques such as single-cell reverse transcriptase-polymerase chain reaction (RT-PCR), offers an exciting new means to identify which biomechanical factors precipitate pathological changes in chondrocytes indicative of osteoarthritis. This article reviews the particular methods used in mechanical studies of single cells with emphasis on techniques that have been used to investigate chondrocytes and similar anchorage-dependent cell types.The fundamentals of RT-PCR and its application at the single-cell level are also discussed. PMID- 12739754 TI - Biomedical engineering analysis of glass impact injuries. AB - This article outlines the history, development, and safety aspects of glass and its use in motor vehicles. It traces the manufacture and describes the characteristics of laminated and tempered glass. It further compares the differences in injuries caused by impact with laminated and tempered glass. The development, use, and results of high penetration resistance (HPR) laminated glass for windshields are examined. Head and neck injuries from impact with glass and glazing structures are delineated. Results of studies with laminated and tempered glass are presented. The probability and severity of injuries occurring secondary to partial or full ejection of vehicle occupants are discussed, and the differences between the performance of laminated and tempered glass are highlighted. Current research to quantify head and neck injury parameters caused by glass impact during rollover is described. The biomechanics of head and neck injury assessment and the development of injury prediction parameters and reference values, respectively, are reviewed. PMID- 12739755 TI - Model-based approaches to biomarker discovery and evaluation: a multidisciplinary integrated review. AB - The most common use of any mathematical or statistical model in physiology, pathophysiology, or therapy evaluation is to organize all relevant components characterizing the system behavior into a rigorously testable framework. This approach is widely applied both to the study of complex homeostatic paradigms involving endogenous substances and to the evaluation of the kinetics and dynamics of xenobiotics such as toxicants and drugs. In either case, one seeks a quantitative framework of the system that is consistent with known physiology and pharmacology and is "compatible" (in some meaningful sense) with all available data. The models are then evaluated and subjected to identifiability and validity tests and can then be used to estimate unknown parameters of interest, to make predictions about system behavior, to simulate previously unobserved behavior in response to a putative perturbation, and to aid in further experimental design. In a broader context, however, the focus and ultimate goal of this set of methodologies (whether this is explicitly stated or not) lies in understanding the mechanisms of physiology and pathophysiology and measuring the effect of therapeutic interventions through the accurate quantification of biomarkers of interest. In this review, we attempt to bring together under this comprehensive framework more than four decades of investigation on modeling and simulation in the life sciences (in particular, we will concentrate on the areas of pharmacology, physiology, and bioengineering). We demonstrate that such modeling approaches, when appropriately designed and evaluated, have significant potential and can be used to understand the multiple factors of disease progression and response to therapeutic interventions, the most likely causes of variability in population and individual responses to therapy, and the most appropriate timing of treatment administration. Lastly, they may allow estimation and prediction of significant outcomes in feasibility and clinical studies. PMID- 12739756 TI - Advances in gastrointestinal electrical stimulation. AB - Electrical stimulation of the gastrointestinal (GI) tract, analogous to pacing the human heart, is an attractive idea. This is because these organs, like the heart, have their own natural pacemakers, and the electrical signals they generate can be altered by externally delivering certain types of electric currents via intraluminal or serosal electrodes to certain areas of the GI tract. A number of studies on animals have been accomplished successfully to treat a variety of disease models, including gastroparesis, dumping, and short bowel syndrome. Over the past 10 years or so, electrical stimulation of the GI tract has received increasing attention among researchers and clinicians because of new techniques, such as implantable devices, and promising results achieved in treatment of gastroparesis and morbid obesity. The objective of this article is to review the advances in electrical stimulation of the gastrointestinal tract. First the electrophysiology of the GI tract and history of GI electrical stimulation are introduced. Then various methods of electrical stimulation of the stomach and small bowel in healthy animals and models of GI diseases are reviewed. Finally clinical applications of electrical stimulation to GI disorders and their possible mechanisms are discussed. PMID- 12739757 TI - Control of multifunctional prosthetic hands by processing the electromyographic signal. AB - The human hand is a complex system, with a large number of degrees of freedom (DoFs), sensors embedded in its structure, actuators and tendons, and a complex hierarchical control. Despite this complexity, the efforts required to the user to carry out the different movements is quite small (albeit after an appropriate and lengthy training). On the contray, prosthetic hands are just a pale replication of the natural hand, with significantly reduced grasping capabilities and no sensory information delivered back to the user. Several attempts have been carried out to develop multifunctional prosthetic devices controlled by electromyographic (EMG) signals (myoelectric hands), harness (kinematic hands), dimensional changes in residual muscles, and so forth, but none ofthese methods permits the "natural" control of more than two DoFs. This article presents a review of the traditional methods used to control artificial hands by means of EMG signal, in both the clinical and research contexts, and introduces what could be the future developments in the control strategy of these devices. PMID- 12739758 TI - Scientific considerations of pharmaceutical solid polymorphism in abbreviated new drug applications. AB - PURPOSE: This commentary is intended to provide a scientific perspective on pharmaceutical solid polymorphism in Abbreviated New Drug Applications (ANDAs). METHODS: This report proposes recommendations for monitoring and controlling drug substance polymorphs and describes scientific considerations of pharmaceutical solid polymorphism in the determination of drug substance sameness. RESULTS: It presents three decision trees for solid oral dosage forms or liquids containing undissolved drug substances to provide a process for evaluating when and how polymorphs of drug substances are monitored and controlled in ANDA submissions. CONCLUSIONS: It is scientifically concluded that differences in polymorphic composition of drug substances in generic drug products and reference-listed drugs are not directly relevant in the determination of drug substance sameness in ANDAs. PMID- 12739759 TI - Fluorescent substrates of sister-P-glycoprotein (BSEP) evaluated as markers of active transport and inhibition: evidence for contingent unequal binding sites. AB - PURPOSE: Although sister-P-glycoprotein (SPGP, BSEP) is closely related to P glycoprotein, it is much more selective in distribution and substrate recognition. Moreover, because inhibition or lack of BSEP function has severe consequences including cholestasis, hepatotoxicity, exposure to toxic xenobiotics, and drug interactions, in vitro methods are necessary for quantifying and characterizing specific inhibition of BSEP. Therefore, the objective is to discern a method and quantitatively characterize several example BSEP inhibitors. METHODS: With fluorescent markers having been used successfully to evaluate and quantify inhibition of P-gp-mediated transport, this study evaluates several compounds for specific cell retention caused by BSEP inhibitors. In addition to the several compounds asserted to be BSEP inhibitors, the compounds suggested to be BSEP substrates might also inhibit BSEP competitively. Retained fluorescence of possible BSEP substrates was measured by a flow cell cytometer using transfected cells presenting the BSEP transporter specifically and abundantly. RESULTS: Several compounds were shown to inhibit BSEP active transport of the fluorescent substrates dihydrofluorescein and bodipy. The inhibition potency was quantified (i.e., cyclosporin A IC50 approximately 7 microM), revealing incongruent relative sensitivities among the substrate markers, with H2FDA generally the most sensitive of the series of substrate markers evaluated. CONCLUSIONS: The inconsistent sensitivities of the transport markers (H2FDA and bodipy) were reminiscent of the apparent multiple binding site behaviors observed for P-gp and could indicate opposing and unequal yet interacting binding sites akin to those of P-gp. Nonetheless, notable differences between P-gp and BSEP in marker substrate recognition/transport were apparent despite the observed overlap in xenobiotic recognition and transport. Thus far the most potent inhibitors seem to be cyclosporin, tamoxifen, and valinomycin. There are likely to be much more potent inhibitors, and other substrates also may be more sensitive to inhibition of transport. PMID- 12739760 TI - 16HBE14o- human bronchial epithelial cell layers express P-glycoprotein, lung resistance-related protein, and caveolin-1. AB - PURPOSE: To study the expression of P-glycoprotein (P-gp), lung resistance related protein (LRP), and caveolin-1 (cav-1) in the human bronchial epithelial cell line 16HBE14o-. METHODS: The presence of P-gp, LRP, and cav-1 in 16HBE14o- cell layers was evaluated using immunocytochemical staining and visualization with confocal laser scanning microscopy (CLSM). Functionality of P-gp was determined by bidirectional transport of rhodamine-123 with and without a P-gp inhibitor, verapamil. Caveolae were visualized using transmission electron microscopy (TEM). Flux of fluorescein-Na was also studied as a paracellular transport marker. RESULTS: Immunocytochemical staining showed expression of P-gp localized at the apical membrane of 16HBE14o- cell layers. The flux of rhodamine 123 across cell layers exhibited a greater Papp value for the secretory (i.e., basolateral-to-apical) direction. This asymmetry disappeared in the presence of verapamil. CLSM provided evidence for the expression of LRP and cav-1. TEM further showed typically shaped caveolae at the apical and basolateral membranes. CONCLUSION: Cell layers of 16HBE14o- express drug transport systems that are also present in the human bronchus in vivo, indicating that the 16HBE14o- cell line may be a suitable candidate for an in vitro model for mechanistic studies of drug transport processes involved in the smaller airways. PMID- 12739761 TI - MDR1 genotype-related duodenal absorption rate of digoxin in healthy Japanese subjects. AB - PURPOSE: Recent clinical studies suggest the importance of the MDR1 genotype at position 3435 (C3435T) in terms of pharmacokinetics, but there is still no consensus in reports on the relationship between the genotype and plasma/serum concentration-time profiles of drugs after conventional oral administration. This study was performed to elucidate the effects of C3435T on the rate of duodenal absorption of digoxin in healthy Japanese subjects. METHODS: Digoxin solution was sprinkled directly over the surface of the duodenum using an endoscope, and its absorption rate was evaluated by serial monitoring of the serum concentration and by analysis of its initial 15-min increasing phase. RESULTS: The duodenal absorption rates of digoxin were 911 +/- 91 ng/min and 506 +/- 76 ng/min for C/C and T/T, respectively (+/- SE, p = 0.007). CONCLUSION: The C3435T mutation of the MDR1 gene is associated with suppression of duodenal absorption of digoxin. PMID- 12739763 TI - Mechanistic studies on nonviral gene delivery to the intestine using in vitro differentiated cell culture models and an in vivo rat intestinal loop. AB - PURPOSE: To identify factors influencing nonviral vector transfection in differentiated CaCo-2 and mucus-secreting coculture, CaCo-2: Ht29GlucH, cell culture models and to compare these in vitro results with in vivo transfection efficiency in rat intestine. METHODS: A range of nonviral vectors including DOTAP, Lipofectin, Superfect, PEI, and polylysine were investigated. CaCo-2 and a mucus-secreting coculture were used at 21 days. Transfection efficiency was assessed using pCMVluc (firefly luciferase) plasmid, and radio-labeled plasmid was used to determine the binding and internalization of plasmid DNA. The in vivo model used was a ligated rat intestinal loop. RESULTS: Transfection levels decreased by over 1000-fold in differentiated models relative to nondifferentiated COS-7 cells and were related to reductions in luciferase production by individual cells. Active internalization of DNA by the differentiated cells decreased. Removal of mucus by the mucolytic agent N acetylcysteine, from the coculture system significantly reduced (p < 0.05) transfection efficiency. In vivo the transfection efficiency of PEI proved superior to DOTAP. CONCLUSIONS: Nonviral gene delivery to the hostile environment of the intestine is possible. Mechanistic studies using differentiated intestinal cell models aid identification of the rate-limiting steps to transfection and represent a more physiologically relevant approach to predict gene delivery to the intestine. PMID- 12739762 TI - Induction of cytochrome P450 2B6 and 3A4 expression by phenobarbital and cyclophosphamide in cultured human liver slices. AB - PURPOSE: To examine the potential of cultured human liver slices to predict cytochrome P450 (CYP) inducibility, regarding global and zonal CYP expression, together with drug-induced histologic changes. METHODS: We first assessed whether CYP2B6, 3A4, and 2C9 expression was maintained in cultured liver slices. Cultured hepatocytes were used as the reference culture system. Then we tested the effects of phenobarbital and cyclophosphamide on CYP expression in both models. RESULTS: Morphologic features are preserved in slices. Basal CYP expression declines with time in culture in both models. Slices display the same region specificity of CYP2B6, 2C9, and 3A4 expression as intact liver. CYP2B6 and 3A4 mRNA, apoprotein, and enzyme-related activities were induced by phenobarbital and cyclophosphamide, whereas CYP2C9 apoprotein was not. Their immunoreactivities were also increased, while their zonal distribution was preserved on slice tissue sections. Microsomal enzyme induction was confirmed by histology. CONCLUSIONS: Cultured human liver slices are an attractive alternative to hepatocyte culture for the prediction of human CYP isoenzyme induction by xenobiotics. PMID- 12739764 TI - Vitreous is a barrier in nonviral gene transfer by cationic lipids and polymers. AB - PURPOSE: To investigate the role of vitreous in nonviral gene delivery into retinal pigment epithelial (RPE) cells. METHODS: Human RPE cell line D407 was cultured in six-well plates. Bovine vitreous, hyaluronan, or DMEM was added on the cells. Complexes of DNA and cationic carriers (polyethyleneimine, poly-L lysine, DOTAP liposomes) were pipetted onto the vitreous, hyaluronan, or DMEM. Cellular uptake of DNA was studied with ethidium monoazide DNA and gene expression with GFP-plasmid complexes. FITC-dextrans and FITC-polylysines were used to probe the effects of the size and cationic charge on permeation in the vitreous in a similar experimental setup. Fluorescent cells were analyzed by flow cytometry. RESULTS: Vitreous decreased the cellular uptake of DNA complexes 2-30 times, and GFP expression was also impaired. In hyaluronan solutions the cellular uptake of the complexes was also decreased significantly in most cases. In vitreous, cellular uptake of all FITC-dextrans decreased slightly, and uptake of poly-L-lysines was decreased substantially, whereas in hyaluronan solutions the effects were mild or nonexistent. CONCLUSIONS: Polymeric and liposomal gene delivery is substantially limited by the vitreous. This is probably because of the size and charge of the retinal gene delivery after intravitreal injections. PMID- 12739765 TI - Influence of preparation conditions on acyclovir-loaded poly-d,l-lactic acid nanospheres and effect of PEG coating on ocular drug bioavailability. AB - PURPOSE: The evaluation of nanosphere colloidal suspensions containing acyclovir as potential ophthalmic drug delivery systems was carried out. The influence of polymer molecular weight and type and concentration of various surfactants on nanosphere properties was studied. The ocular pharmacokinetics of acyclovir loaded nanoparticles was evaluated in vivo and compared with an aqueous suspension of the free drug. METHODS: Nanospheres were made up of poly-d,l-lactic acid (PLA). The colloidal suspension was obtained by a nanoprecipitation process. The surface properties of PLA nanospheres were changed by the incorporation of pegylated 1,2-distearoyl-3-phosphatidylethanolamine. The mean size and zeta potential of the nanospheres were determined by light scattering analysis. The acyclovir loading capacity and release were also determined. In vivo experiments were carried out on male New Zealand rabbits. The ocular tolerability of PLA nanospheres was evaluated by a modified Draize test. The aqueous humor acyclovir levels were monitored for 6 h to determine the drug's ocular bioavailability for the various formulations. RESULTS: A reduction of the mean size and a decrease of the absolute zeta potential of PLA nanospheres resulted from increasing the surfactant concentration. The higher the polymer molecular weight, the smaller the nanosphere mean size. PEG-coated and uncoated PLA nanospheres showed a sustained acyclovir release and were highly tolerated by the eye. Both types of PLA nanospheres were able to increase the aqueous levels of acyclovir and to improve the pharmacokinetics profile, but the efficacy of the PEG-coated nanospheres was significantly higher than that of the simple PLA ones. CONCLUSIONS: PEG-coated PLA nanospheres can be proposed as a potential ophthalmic delivery system for the treatment of ocular viral infections. PMID- 12739766 TI - How well can a T-cell epitope replace its parent carrier protein? A dose-response study. AB - PURPOSE: This work examines the effectiveness of synthetic peptide immunogens derived from immunodominant T-cell epitopes as replacements for their intact parent protein in vaccines. METHODS: Fluorescein was conjugated to hen egg lysozyme (FL-HEL, positive control) and three synthetic peptide immunogens: (a) murine B10.A (H-2a) immunodominant T-cell epitope of HEL [FL-(T-cell epitope)]; (b) multiple antigenic peptide (MAP) multimer of this epitope ([FL-(T epitope)]n MAP, n = 2-4); and (c) negative control MAP with T-cell epitope residues replaced with glycine [(FL-Gly18)4-MAP]. The dose response of each immunogen was examined over a 300-fold range in B10.A mice. The immune response was monitored using antifluorescein ELISA assays. RESULTS: FL-(T epitope)'s immune response correlated positively with dose, with maximum response comparable to that of [FL (T epitope)]n-MAP, or FL-HEL. This trend was consistent across 1 degrees, 2 degrees, and 3 degrees responses, although interanimal variability was higher in the latter two because of an all-or-none response in mice immunized with this peptide. [FL-(T epitope)]n-MAP's immune response was consistently high and nearly dose independent, a trend observed across 1 degrees, 2 degrees, and 3 degrees responses. FL-HEL's immune response correlated negatively to dose in the 1 degrees response but was nearly dose independent in the 2 degrees and 3 degrees responses. The magnitude of these latter responses was comparable to that observed for [FL-(T epitope)]n-MAP. (FL-Gly18)4-MAP did not elicit an immune response except at the highest dose. This trend was consistent across 1 degrees, 2 degrees, and 3 degrees responses. CONCLUSIONS: The monomeric epitope was 300 fold less potent than its parent carrier protein, but increasing immunogen valency using MAP technology compensated totally for reduced potency. (FL-Gly18)4 MAP's lack of response at all but the highest dose strongly suggests that a specific immunodominant T-cell epitope sequence for HEL is necessary for successful peptide mimicry of HEL. This work also demonstrates the importance of quality assessment of commercial MAP core resins. PMID- 12739767 TI - Contribution of ion-pair complexation with bile salts to the transport of organic cations across LLC-PK1 cell monolayers. AB - PURPOSE: To examine the effect of ion-pair complexation with endogenous bile salts on the transport of organic cations (OCs) across LLC-PK1 cell monolayers. METHODS: The transport of tributylmethyl-ammonium (TBuMA) and triethylmethylammonium (TEMA) across the cell monolayer was measured in the presence of taurodeoxycholate (TDC), an endogenous organic anion that forms an ion-pair complex with TBuMA, but not with TEMA. RESULTS: Under proton gradient conditions (i.e., pH 6.0 apical/pH 7.4 basal), the above OCs exhibited similar transport charactersistics, consistent with the well-established OC/H+ antiporter, and the presence of TDC had no measurable effect on the transport of these OCs. Under pH-equilibrated conditions (i.e., pH 7.4 apical/pH 7.4 basal); however. basal to apical transport of TBuMA, not that of TEMA, was increased in the presence of TDC, probably as a result of the formation of a lipophilic ion pair complex between TBuMA and TDC. The transport and efflux of the TBuMA-TDC complex across the apical membrane of the cell was inhibited by representative substrates of the P-glycoprotein (P-gp), indicating the involvement of P-gp in this process. The increased affinity of the ion-pair complex to P-gp is consistent with a mechanism involving increased transport. CONCLUSION: In cases where there is no proton gradient between the plasma and urine, the formation of lipophilic ion-pair complexes in the kidney with endogenous bile salts might be involved in the in vivo urinary excretion of large Mw OCs, such as TBuMA. PMID- 12739768 TI - Development of a dog microdialysis model for determining synovial fluid pharmacokinetics of anti-arthritis compounds exemplified by methotrexate. AB - PURPOSE: The purpose of this study was to develop and validate an animal model of drug disposition in synovial fluid (SF) by comparing microdialysis with arthrocentesis using the anti-arthritic drug methotrexate (MTX). METHODS: Microdialysis probes were calibrated in vitro with the no net flux method using dog synovial fluid. The probes were implanted surgically into the stifle joint space of four dogs and were dialyzed overnight using a portable microinfusion pump. The membrane integrity of the probes was monitored by retrodialysis using an internal standard. After an intravenous bolus of 2.5 mg/kg of MTX, unbound concentrations in synovial fluid, as well as total plasma concentrations, were measured by liquid chromatography tandam mass spectrometer (LC/MS/MS) in samples collected from 0 to 48 h postdose. RESULTS: The probe membrane remained intact at least 48 h after implantation. The mean probe recovery and unbound fraction of MTX in SF were 46.8% and 44.8%, respectively. The unbound fraction of MTX was 44% in synovial fluid. MTX penetrated into the joint space rapidly, with maximal concentrations of 6.6 microM reached at approximately 1 h postdose. The unbound MTX area under the curve in SF was approximately 40% of the total area under the curve in plasma. These data agree well with the previous data obtained for MTX using arthrocentesis. CONCLUSION: In contrast with arthrocentesis, microdialysis enables the collection of multiple serial SF samples from individual animals with minimal trauma and potential blood contamination. This animal model should prove valuable for studying the disposition of new antiarthritis compounds or biomarkers in SF. PMID- 12739769 TI - Evaluation of inhibitory activity of casein on proteases in rat intestine. AB - PURPOSE: To investigate the possible use of casein as an adjuvant for oral delivery of peptide drugs, the inhibitory activity of casein on proteases in rat small intestine was examined. METHODS: Male Sprague-Dawley rats weighing 200-300 g were used as the animal model. The luminal contents of the small-intestinal tract and mucosal subcellular fractions of the small intestine were prepared: the enzymatic activities of trypsin, chymotrypsin, aminopeptidase-B, leucine aminopeptidase, dipeptidylaminopeptidase-IV, cathepsin B, and dipeptidylaminopeptidase-II were determined using a specific substrate for each protease; and the effect of casein on the protease activity was examined. RESULTS: Casein strongly inhibited trypsin and chymotrypsin in the concentration dependent manner. As to the proteases in the intestinal epithelial cells, casein inhibited an endopeptidase, cathepsin B, but not other exopeptidases. The inhibitory activity was independent of the type of casein. The kinetic analysis characterized the type of inhibition on trypsin and chymotrypsin to be competitive. CONCLUSIONS: Casein was shown to have strong inhibitory activity on trypsin and chymotrypsin in the intestinal lumen. Taken into consideration that trypsin and chymotrypsin are endopeptidases, it is suggested that casein has strong inhibitory activity only on endopeptidases. PMID- 12739770 TI - Morphine blood-brain barrier transport is influenced by probenecid co administration. AB - PURPOSE: The objective of this study was to investigate the possible influence of probenecid on morphine transport across the blood-brain barrier (BBB) in rats. METHODS: Microdialysis probes, calibrated using retrodialysis by drug, were placed into the striatum and jugular vein of seven Sprague-Dawley rats. Morphine was administered as a 4-h exponential infusion. The experiment was repeated the next day with the addition of probenecid, administered as a bolus dose (20 mg/kg) followed by a constant infusion (20 mg/kg/h). Models for BBB transport were built using the computer program NONMEM. RESULTS: The steady-state ratio of 0.29 +/- 0.07 of unbound morphine concentration in brain to that in blood indicates that morphine is actively effluxed at the BBB. Probenecid co-administration increased the ratio to 0.39 +/- 0.04 (p < 0.05). Models in which probenecid influenced the brain efflux clearance rather than the influx clearance, well described the data. The half-life in brain increased from 58 +/- 9 min to 115 +/- 25 min when probenecid was co-administered. Systemic clearance of morphine also decreased upon probenecid co-administration, and M3G formation was decreased. CONCLUSION: This study indicates that morphine is a substrate for the probenecid-sensitive transporters at the BBB. Co-administration of probenecid decreased the brain efflux clearance of morphine. PMID- 12739771 TI - Cell handling, membrane-binding properties, and membrane-penetration modeling approaches of pivampicillin and phthalimidomethylampicillin, two basic esters of ampicillin, in comparison with chloroquine and azithromycin. AB - PURPOSE: The purpose of this work was to examine and understand the cellular pharmacokinetics of two basic esters of ampicillin, pivaloyloxymethyl (PIVA) and phthalimidomethyl (PIMA), in comparison with lysosomotropic drugs (chloroquine, azithromycin). METHODS: Cell culture studies (J774 macrophages) were undertaken to study uptake and release kinetics and to assess the influence of concentration, pH, proton ionophore (monensin), and MRP and P-gp inhibitors (probenecid, gemfibrozil, cyclosporin A, GF 120918). Equilibrium dialysis with liposomes were performed to directly asses the extent of drug binding to bilayers. Conformational analysis modeling of the drug penetration in bilayers was conducted to rationalize the experimental observations. RESULTS: PIVA and PIMA showed properties in almost complete contrast with those of chloroquine and azithromycin, i.e., fast apparent accumulation and fast release at 4 degrees C as well as at 37 degrees C, saturation of uptake (apparent Kd 40 microM), no influence of monensin, MRP, or P-gp inhibitors; tight binding to liposomes (Kd approx. 40 microM); and sharp increase in calculated free energy when forced in the hydrophobic domain. CONCLUSIONS: Although they are weak organic bases, PIVA and PIMA show none of the properties of lysosomotropic agents. We hypothesize that they remain locked onto the pericellular membrane and may never penetrate cells as such in significant amounts. PMID- 12739773 TI - Topical delivery of 5-fluorouracil and 6-mercaptopurine by their alkylcarbonyloxymethyl prodrugs from water: vehicle effects on design of prodrugs. AB - PURPOSE: To determine whether the fluxes through hairless mouse skin for three homologous series of prodrugs of 5-fluorouracil (5-FU, 1) and 6-mercaptopurine (6 MP, 2) from saturated aqueous suspensions show dependencies on aqueous (SAQ) and isopropyl myristate (SIPM) solubilities similar to those shown by the identical compounds delivered from IPM. METHODS: Flux through hairless mouse skin from water (JMAQ) and solubility data were measured for a homologous series of six 3 alkylcarbonyloxymethyl (ACOM) prodrugs of 5-FU (3-ACOM-5-FU), and five 6-ACOM-6 MP prodrugs, then combined with literature data for five bis-6,9-ACOM-6-MP prodrugs to give a data base. Multiple linear regression using SPSS 7.5 was performed on log SIPM, log SAQ, molecular weight and log JMAQ data to determine the best fit coefficients to the transformed Potts-Guy equation: log JMAQ = x + y log SIPM + (1 - y) log SAQ + z MW. Permeability coefficients (PMAQ) were calculated from JMAQ/SAQ. RESULTS: The best fit coefficients for the flux from AQ(JMAQ) were x = -1.497, y = 0.660 and z = -0.00469 (r2 = 0.765) with an average error of prediction equal to 0.193 log units. The best fit coefficients for the flux from IPM (JMIPM) were x = -0.557, y = 0.536 and z = -0.00261 (r2 = 0.941) with an average error of prediction equal to 0.109 log units. For all three series, log PMAQ increased whereas log PMIPM decreased with increasing alkyl chain lengths in the promoiety and with decreasing solubility parameter values. CONCLUSIONS: The transformed Potts-Guy equation can be used to predict JMAQ but with less certainty than JMIPM. SIPM and SAQ have consistently been shown to have a positive influence on JMIPM, and now on JMAQ, with a balance between the two solubilities being obviously important. The previous observation that log PMAQ increased with lipophilicity is an artifact of normalizing JMAQ by SAQ. PMID- 12739772 TI - Noninvasive characterization of regional variation in drug transport into human stratum corneum in vivo. AB - PURPOSE: To investigate the mechanisms underlying the regional variations in drug transport into human stratum corneum (SC) of two model compounds of different lipophilicity and molecular size, 4-cyanophenol (CP) and cimetidine (CM), in vivo by non-invasive, quantitative attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy. METHODS: Saturated solutions of CP and CM were applied to the skin surface of eleven Chinese men, at five anatomical sites, including forearm, back, thigh, leg, and abdomen, for 10-15 min and 3-5 h, respectively. After the skin surface was cleansed of remaining chemicals, the SC was tape-stripped sequentially up to 20 times, and the drug concentration profiles in the tape-stripped SC were determined using ATR-FTIR spectroscopy. Thickness of the SC was estimated simultaneously using two-point measurements of transepidermal water loss before and after completion of tape stripping. Estimation of partition, diffusion, and permeability coefficients was achieved by analysis of the data using the unsteady-state diffusion equation. RESULTS: The rank orders of regional variation in partition and diffusion coefficients of CP and CM were different. The rank order of regional variation in permeability coefficients was similar for both drugs and decreased in the order of back > forearm > thigh > leg > or = abdomen, but the variation was more prominent for CM. CONCLUSIONS: Regional variation in SC transport of CP was mainly influenced by its intrinsic diffusivity across the SC, whereas variation in transport of CM could be attributed to both thermodynamic and kinetic differences among different anatomical skin sites. PMID- 12739774 TI - Electroosmotic pore transport in human skin. AB - PURPOSE: To determine the pathways and origin of electroosmotic flow in human skin. METHODS: Iontophoretic transport of acetaminophen in full thickness human cadaver skin was visualized and quantified by scanning electrochemical microscopy. Electroosmotic flow in the shunt pathways of full thickness skin was compared to flow in the pores of excised stratum corneum and a synthetic membrane pore. The penetration of rhodamine 6G into pore structures was investigated by laser scanning confocal microscopy. RESULTS: Electroosmotic transport is observed in shunt pathways in full thickness human skin (e.g., hair follicles and sweat glands), but not in pore openings of freestanding stratum corneum. Absolute values of the diffusive and iontophoretic pore fluxes of acetaminophen in full thickness human skin are also reported. Rhodamine 6G is observed to penetrate to significant depths (approximately 200 microm) along pore pathways. CONCLUSIONS: Iontophoresis in human cadaver skin induces localized electroosmotic flow along pore shunt paths. Electroosmotic forces arise from the passage of current through negatively charged mesoor nanoscale pores (e.g., gap functions) within cellular regions that define the pore structure beneath the stratum corneum. PMID- 12739775 TI - Pretreatment with a water-based surfactant formulation affects transdermal iontophoretic delivery of R-apomorphine in vitro. AB - PURPOSE: To further increase the transdermal transport rate of R-apomorphine, a nonocclusive pretreatment with an aqueous surfactant formulation in combination with iontophoresis was explored in vitro. METHODS: The human stratum corneum was pretreated nonocclusively with formulations composed of laureth-3 oxyethylene ether (C12EO3), laureth-7 oxyethylene ether (C12EO7), and cholesterol sulfate (CSO4) prior to iontophoresis. The effect on the flux of the following parameters was examined: the composition, the charge, and the applied amount of surfactant formulations. RESULTS: The iontophoretic flux of R-apomorphine was appreciably increased by pretreatment with surfactant formulations. A formulation containing C12EO3/C12EO7/CSO4 at a molar ratio of 70:30:5 was very stable and increased the iontophoretic flux of R-apomorphine from 92.2 +/- 13.9 nmol/cm2 x h to 181.5 +/- 22.6 nmol/cm2 x h. When further increasing the negative charge of this formulation the iontophoretic transport rate was slightly inhibited. A dose of 40 microL/cm2 of the formulation with a total surfactant concentration of 5% (w/w) was sufficient for a maximum enhancing effect. CONCLUSIONS: The results obviously show that nonocclusive pretreatment with the surfactant formulation enhances the iontophoretic transport of R-apomorphine, and is a promising approach to achieve therapeutic concentrations of R-apomorphine. PMID- 12739776 TI - Effective inhibition of mannitol crystallization in frozen solutions by sodium chloride. AB - PURPOSE: The purpose of this work was to study the possibility of preventing mannitol crystallization in frozen solutions by using pharmaceutically acceptable additives. METHODS: Differential scanning calorimetry (DSC) and low-temperature X ray diffractometry (LTXRD) were used to characterize the effect of additives on mannitol crystallization. RESULTS: DSC screening revealed that salts (sodium chloride, sodium citrate, and sodium acetate) inhibited mannitol crystallization in frozen solutions more effectively than selected surfactants, alpha cyclodextrin, polymers, and alditols. This finding prompted further studies of the crystallization in the mannitol-NaCl-water system. Isothermal DSC results indicated that mannitol crystallization in frozen solutions was significantly retarded in the presence of NaCl and that NaCl did not crystallize until mannitol crystallization completed. Low-temperature X-ray diffractometry data showed that when a 10% w/v mannitol solution without additive was cooled at 1 degrees C/min, the crystalline phases emerging after ice crystallization were those of a mannitol hydrate as well as the anhydrous polymorphs. In the presence of NaCl (5% w/v), mannitol crystallization was suppressed during both cooling and warming and occurred only after annealing and rewarming. In the latter case however, mannitol did not crystallize as the hydrate, but as the anhydrous delta polymorph. At a lower NaCl concentration of 1% w/v, the inhibitory effect of NaCl on mannitol crystallization was evident even during annealing at temperatures close to the Tg' (-40 degrees C). A preliminary lyophilization cycle with polyvinyl pyrrolidone and NaCl as additives rendered mannitol amorphous. CONCLUSION: The effectiveness of additives in inhibiting mannitol crystallization in frozen solutions follows the general order: salts > alditols > polyvinyl pyrrolidone > alpha-cyclodextrin > polysorbate 80 approximately polyethylene glycol approximately poloxamer. The judicious use of additives can retain mannitol amorphous during all the stages of the freeze-drying cycle. PMID- 12739777 TI - Stability and release performance of a series of pegylated copolymeric micelles. AB - PURPOSE: The aim of this work is to evaluate the capability of a series of biocompatible amphiphilic copolymers as a nano-sized drug carrier. METHODS: The influences of the type of lactone monomer, the feed molar ratios of lactone/PEG, and the molecular weight of PEG on the performance and release behavior of micelles are investigated. RESULTS: These pegylated amphiphilic copolymers efficiently form micelles with a low CMC value in the range of 10(6)-10(-7) M. The average particle size of micelles is approximately 100 nm. The phenomenon of increasing particle size as increasing the chain length of poly(lactone) block is observed. The different hydrophobicity, based on chemical structure of poly(lactone), accounts for different interaction strength between indomethacin and hydrophobic inner core, which further influences the drug loading in copolymeric micelles and their release character. In addition, the PCL/PEG/PCL micellar solutions maintain their sizes at 4 degrees C for 8 weeks without occurring significant aggregation or dissociation. CONCLUSIONS: A series of biocompatible pegylated amphiphilic copolymers have been elucidated possessing micellization potential to form nano-sized micelles in an aqueous environment, which enable incorporate hydrophobic drug and regulate drug release. PMID- 12739778 TI - The stenlying effect of high hydrostatic pressure on thermally and hydrolytically labile nanosized carriers. AB - PURPOSE: To investigate whether high hydrostatic pressure (HHP) treatment allows the sterilization of thermosensitive polymer nanoparticle suspensions without jeopardizing their physicochemical integrity. METHODS: Application of HHP was explored on a wide variety of thermosensitive poly(cyanoacrylate) nanoparticles, varying by their type (nanospheres or nanocapsules), by their preparation method (nanoprecipitation or emulsion/solvent evaporation), as well as by their surface characteristics. Physicochemical characterization before and after pressurization included turbidimetry, size measurement, zeta potential, scanning electron microscopy and infrared analysis. A sterility test also conducted according to pharmacopoeial requirements on an importantly contaminated nanoparticle suspension. RESULTS: Poly(cyanoacrylate) nanoparticles appeared to be extremely baroresistant. Continuous or oscillatory HHP treatment up to 500 MPa during 30 min induced generally neither physical, nor chemical damage. However, precautions should be taken when surface modifiers are adsorbed onto nanoparticles, as a layer destabilization may occur. Finally, this process allowed the successful inactivation of vegetative bacteria, yeast, and fungi. CONCLUSIONS: This work proposes HHP as a new method for polymer drug carriers sterilization, taking into account that further exploration in this area is needed to propose novel protocols for spores inactivation. PMID- 12739780 TI - Correlation of rFVIII inactivation with aggregation in solution. AB - PURPOSE: This study was designed to investigate the stability of recombinant FVIII (rFVIII) in solution at different pHs and to probe the cause(s) of rFVIII inactivation under accelerated storage conditions. METHODS: Aqueous stability samples of full-length rFVIII at different pHs were incubated at 40 degrees C for several days and analyzed by the one-stage clotting assay. SEC-HPLC, SDS-PAGE, and UV spectrophotometry. RESULTS: Incubation of liquid rFVIII at 40 degrees C inactivated the protein rapidly and linearly with time on a semi-log scale at all pHs, suggesting a first order or pseudo first order process. A U-shaped relationship was found between the rate constant for loss of rFVIII activity and the solution pH. The minimal rate of inactivation was found between pH 6.6 and 7.0 with a half-life of approximately 4 days. The SEC-HPLC results indicated pH dependent aggregation of rFVIII during incubation. It was found that the disappearance of monomeric rFVIII by SEC-HPLC correlated with the loss of rFVIII activity (r2 = 0.97). Both the SDS-PAGE and UV results confirmed the aggregation pathway of rFVIII. In addition, the SDS-PAGE results suggest involvement of three aggregation mechanisms--disulfide-bond formation/exchange, non-reducible crosslinking, and physical interactions. CONCLUSIONS: The full-length rFVIII is unstable in solution at 40 degrees C and loses activity rapidly through a first order or pseudo first order aggregation process, which consists of both physical and chemical pathways. SEC-HPLC may be used in monitoring rFVIII stability studies in lieu of the clotting assay under the incubation conditions used in this study. PMID- 12739781 TI - From the Big Bang to B27. PMID- 12739779 TI - Validation of the chloramine-T induced oxidation of human serum albumin as a model for oxidative damage in vivo. AB - PURPOSE: The validity of using chloramine-T as a model compound for mimicing oxidative stress was examined using human serum albumin (HSA) as a model. Important sites of oxidation were studied by mild treatment with chloramine-T and by mutating 34Cys for a serine (C34S). METHODS: High-performance liquid chromatography (HPLC) combined with fluorescence detection to confirm the validity of chloramine-T as an oxidizing agent was used. Oxidized amino acid residues were detected by reaction with 5,5'-dithiobis(2-nitro benzoic acid), digestion with cyanogen bromide, followed by capillary electrophoresis. Protein conformation was examined by spectroscopic techniques. RESULTS: From the HPLC analysis of human serum, the validity of using chloramine-T as an oxidizing agent was confirmed. At low chloramine-T concentrations (CT0.1-HSA, CT1-HSA), 34Cys and Met residues were oxidized, at medium concentrations (CT10-HSA), the tryptophan residue also appeared to be oxidized, and at the highest concentration (CT50 HSA), the net charge of Site II of HSA was found to be more negative. The two highest levels of oxidation of HSA (CT10-HSA, CT50-HSA) resulted in conformational changes with an increased exposure of hydrophobic regions, decreased high-affinity bindings of warfarin and ketoprofen and a reduced esterase-like activity. The latter protein also has a shorter plasma half-life and an increased liver clearance. CONCLUSIONS: We succeeded in imitating oxidative damage to HSA using chloramine-T and the findings show that Site II is more affected than Site I and 34Cys, when HSA is exposed to oxidative stress. PMID- 12739782 TI - Oral tolerance and pyruvate dehydrogenase in patients with primary biliary cirrhosis. AB - Primary biliary cirrhosis (PBC) is a chronic cholestatic liver disease characterized by the immunological destruction of intralobular bile ducts and serum anti-mitochondrial antibodies (AMA). Based upon previous work of oral tolerance and autoimmunity, we hypothesized that feeding the mitochondrial autoantigens of PBC would alter the clinical course and the level of antimitochondrial antibodies. The bovine pyruvate dehydrogenase complex (PDC) was purified and 5 mg fed in gelatin capsules to 6 patients with early stage PBC for 6 months. Antimitochondrial antibodies and liver biochemistries were measured at every 3 months for 12 months. The clinical trial was completed for all patients except for 1 who showed deterioration of pre-existing skin rash during treatment, which disappeared within 2 weeks after treatment was discontinued. However, after 1 year, neither the titers of AMAs nor liver biochemistries were significantly changed by this treatment. This is the first trial to test the efficacy of oral tolerance induction in PBC. However, the data, which limited in scope, did not demonstrate efficacy and further highlights the difficulties in showing continuing evidence of tolerance induction in autoimmunity. PMID- 12739783 TI - Different roles of a rat cortical thymic epithelial cell line in vitro on thymocytes and thymocyte hybridoma cells: phagocytosis, induction of apoptosis, nursing and growth promoting activities. AB - In this work, the interaction between a rat cortical thymic epithelial cell (TEC) line (R-TNC.1) with nursing activity and thymocytes as well as BWRT 8 thymocyte hybridoma (TH) cells has been studied. The R-TNC.1 cell line significantly bound thymocytes and TH. Binding was stronger during the first 30 min of cell incubation and was followed by a progressive deadhesion. Among adherent thymocytes the proportion of apoptotic cells increased with culture time which was a consequence of higher capacity of the line for binding of apoptotic than viable cells and induction of apoptosis in a subset of adherent thymocytes. Emperiopolesis activity of this thymic nurse cell (TNC) line was manifested by engulfment of thymocytes as well as TH cells. A subset of viable intra-TNC thymocytes has been triggered to die by apoptosis, whereas other internalized thymocytes have been stimulated to proliferate, as measured by an increase in the percentage of cells in mitosis and higher incorporation of bromodeoxyuridine (BrdU), in comparison to thymocytes cultivated alone. A significant stimulation of proliferation of engulfed TH cells was also observed. The R-TNC.1 cell line efficiently phagocytosed both apoptotic thymocytes and TH, and the process is followed by intra-TNC destruction of ingested cells. Cumulatively, these results suggest different role of the R-TNC.1 clone: phagocytosis of apoptotic cells; induction of apoptotic cell death in a subset of both bound and internalized thymocytes and stimulation of proliferation of a subset of intra-TNC thymocytes or TH cells. PMID- 12739784 TI - Activated alpha4 integrins are preferentially expressed on immature thymocytes and activated T cells. AB - We have identified a novel mAb, SG31, which recognizes the mouse integrin alpha4 subunit. Unlike the epitopes recognized by other anti-alpha4 antibodies, the SG31 epitope is expressed on subpopulations of thymocytes and peripheral T cells. After manganese ion, but not phorbol myristic acetate activation, the epitope is induced and expressed on the majority of peripheral T cells. These data suggest that the SG31 epitope is an activation epitope and that manganese ions activate alpha4 integrins by inducing a conformational change. Comparative flow cytometric analyses showed that the SG31 epitope as well as the epitope detected by other anti-alpha4 antibodies is expressed on all B lineage cells. In the T lineage, expression of the alpha4 integrins is down-regulated during thymocyte development. Although mature thymocytes still express the alpha4 integrins, they lose almost entirely the activation epitope recognized by SG31. In contrast, the most immature thymocytes express high levels of this epitope. In the periphery, SG31 epitope is expressed mostly by activated T cells, in contrast to the overall population of T cells that express the alpha4 integrins at homogenous levels. These results suggest that the activation of the alpha4 integrins is parallel to that of T cells. PMID- 12739785 TI - Differential regulation of mouse B cell development by transforming growth factor beta1. AB - Transforming growth factor beta (TGFbeta) can inhibit the in vitro proliferation, survival and differentiation of B cell progenitors, mature B lymphocytes and plasma cells. Here we demonstrate unexpected, age-dependent reductions in the bone marrow (BM) B cell progenitors and immature B cells in TGFbeta1-/- mice. To evaluate TGFbeta responsiveness during normal B lineage development, cells were cultured in interleukin 7 (IL7) +/- TGFbeta. Picomolar doses of TGFbeta1 reduced pro-B cell recoveries at every timepoint. By contrast, the pre-B cells were initially reduced in number, but subsequently increased compared to IL7 alone, resulting in a 4-fold increase in the growth rate for the pre-B cell population. Analysis of purified BM sub-populations indicated that pro-B cells and the earliest BP1- pre-B cells were sensitive to the inhibitory effects of TGFbeta1. However, the large BP1+ pre-B cells, although initially reduced, were increased in number at days 5 and 7 of culture. These results indicate that TGFbeta1 is important for normal B cell development in vivo, and that B cell progenitors are differentially affected by the cytokine according to their stage of differentiation. PMID- 12739786 TI - Developmental considerations of sperm protein 17 gene expression in rheumatoid arthritis synoviocytes. AB - Rheumatoid arthritis (RA) is an autoimmune disease characterized by proliferative synovial tissue. We used mRNA differential display and library subtraction to compare mRNA expression in RA and osteoarthritis (OA) synoviocytes. We initially compared the mRNA expression patterns in 1 female RA and 1 OA synovia and found a differentially expressed 350 bp transcript in the RA synoviocytes which was, by sequence analysis, 100% homologous to sperm protein 17 (Sp17). Moreover, the Sp17 transcript was found differentially expressed in a RA synovial library that was subtracted with an OA synovial library. Using specific primers for full length Sp17, a 1.1 kb transcript was amplified from the synoviocytes of 7 additional female RA patients, sequenced and found to 100% homologous to Sp17. Thus, we found the unexpected expression of Sp17, a thought to be gamete-specific protein, in the synoviocytes of 8/8 female RA patients in contrast to control OA synoviocytes. Interestingly, Sp17's structural relationship with cell-binding and recognition proteins, suggests that Sp17 may function in cell-cell recognition and signaling in the RA synoviocyte. Further, Sp17 could have a significant regulatory role in RA synoviocyte gene transcription and/or signal transduction. Thus, Sp17 could have an important role in RA synoviocyte proliferation or defective apoptosis. Finally, the presence of Sp17 in synoviocytes has interesting developmental considerations. PMID- 12739788 TI - Sex-related health risks and implications for interventions with hospitality women in Hainan, China. AB - This article describes the living situations of sex workers (hospitality girls) in a typical rural town in southern China; their personal, social, financial, and psychological needs and expectations; and the social and economic factors that place them at risk for STIs/HIV and unwanted pregnancy, based on the qualitative and quantitative data collected prior to and during a preventive intervention. Though some similarities to knowledge, attitudes, and behaviors of sex workers reported in other Asian countries were identified, this study identified some differences distinctive to the China context, such as how economic factors, kinship, and friendship networks function in the service industry, and how the culture of reproduction and birth control policy interact with and affect sex workers' health and decision-making options. Understanding these factors is critical for design of a culturally and contextually tailored intervention for the reduction of sex-related health risks of women in the sex service industry. PMID- 12739787 TI - Autoimmune cholangitis in the SJL/J mouse is antigen non-specific. AB - Primary biliary cirrhosis (PBC) is an autoimmune disease characterized by intrahepatic bile duct destruction and the production of anti-mitochondrial antibodies (AMA). The absence of an animal model has been a striking impedance in defining the molecular basis of disease. Previous work has suggested that SJL/J mice immunize with the pyruvate dehydrogenase complex (PDC-E2), the major mitochondrial autoantigen of PBC, leads to the development of lymphoid cell infiltration in portal tracts and a model system coined autoimmune cholangitis. We hypothesized that this pathology would be augmented if immunization occurred in the presence of IFN-gamma injections. Accordingly, SJL/J mice were immunized with PDC-E2 and, for purpose of control, alpha-casein. Subgroups of mice were also treated with exogenous IFN-gamma. As expected, mice immunized with PDC-E2, with or without IFN-gamma, developed high titer AMAs. In contrast, mice immunized with alpha-casein, develop antinuclear antibodies. More importantly, the livers from mice immunized with PDC-E2 and/or those immunized with alpha-casein all displayed lymphoid cell infiltration to the portal tracts, irrespective of bile duct size. Indeed, there was no significant difference between the experimental and the control groups by histologic analysis. Thus, autoimmune cholangitis in these mice is antigen non-specific. PMID- 12739789 TI - Technological tearoom trade: characteristics of Swedish men visiting gay Internet chat rooms. AB - This study compares differences among Swedish men who never, occasionally, and frequently use Internet sexual chat rooms. The data indicate that Internet sexual chat room users are significantly different from those who never visit chat rooms. The users were younger, more likely to live at home or with a female partner, bisexual, less open about their homosexuality, less likely to be members of gay organizations, and more likely to engage in unprotected anal intercourse with casual partners. The Internet might be a mean's of approximating homosexual contact. These data suggest that the Internet may be a useful place to reach younger and bisexual men, and those who make sexual assignations, with HIV/STD preventive messages, often before they have publicly come out. PMID- 12739790 TI - Binge use of methamphetamine among HIV-positive men who have sex with men: pilot data and HIV prevention implications. AB - This study compared the social and behavioral characteristics of binge users and nonbinge users of methamphetamine (meth) in a sample of 90 HIV-positive men who have sex with men. Forty-one participants (46%) self-identified as a binge user. Meth binges ranged from 2 to 33 days (mean = 5.6), and average consumption was 3.1 grams. Binge users were significantly more likely than nonbinge users to be ethnic minority and to have lower education. The two groups did not differ in terms of the total amount of meth used in the past 30 days; however, binge users reported significantly more social difficulties, more mental and physical health problems, and more sexual risk behaviors as compared with nonbinge users. The findings are discussed in relation to drug treatment approaches and the development of behavioral interventions. PMID- 12739792 TI - Differences in altruistic roles and HIV risk perception among staff, peer educators, and students in an adolescent peer education program. AB - This qualitative study examines how adolescent peer educators understand and communicate HIV prevention messages. Semistructured ethnographic interviews were conducted with 21 program participants, including staff, peer educators, and students. Interviews were transcribed and analyzed using concept analysis, a method for identifying shared concepts among interview subjects. We found (a) similar beliefs about HIV transmission and risk reduction across groups; (b) different, but strong, altruistic roles among staff and peer educators; and (c) differences in HIV risk perception across the three groups. Altruistic roles took two forms. Staff acted as life skills mentors, whereas peer educators acted as HIV educators. Students were more passive, receiving counseling but not passing it on to others. Staff contextualized HIV risk, whereas peer educators and students emphasized risk. Although similar HIV knowledge across groups suggests program efficacy, stronger altruistic roles or contextualization of HIV risk may affect how prevention messages are delivered. PMID- 12739791 TI - Developing a culturally appropriate video to promote dual-method use by urban teens: rationale and methodology. AB - This article describes the development and production of a video targeted to urban adolescent women who use hormonal contraception. Research has shown that this population is at high risk for nonuse of condoms. Extensive preliminary qualitative and quantitative studies were done to understand the situations in which young women do and do not ask their partners to use a condom. Combining constructs from the health belief model, social cognitive learning theory and the theory of reasoned action with insights gleaned from the preliminary studies, the researchers developed a video that features two young HIV-positive women and two young HIV-negative men. Focus groups with young women helped the researchers edit the video to its current length of 21 minutes. Responses from the focus groups suggest that the video has a strong impact on young women. The video is currently part of a randomized controlled trial of two HIV prevention interventions. PMID- 12739794 TI - Examining strategies for culturally grounded HIV prevention: a review. AB - Since the early 1990s culture has been considered an essential concept for understanding the HIV/AIDS epidemic and the prevention of HIV (Parker, 2001). Despite consensus that culture is an important issue in HIV prevention programming, the field lacks a common vision for how culture ought to inform intervention design and implementation. In this article, we review the HIV prevention literature published through 2001 to examine interventions that have explicitly sought to address cultural concepts. We describe the types of strategies used to integrate culture into HIV prevention, how culture has been evaluated as a component of preventive interventions, and to whom these culturally grounded programs are targeted. We highlight gaps in the current body of literature and provide recommendations for future research on culturally grounded HIV prevention programs. PMID- 12739793 TI - Do community planning groups (CPGs) influence HIV prevention policy? An analysis of California CPGs. AB - Since 1994, the Centers for Disease Control and Prevention has required its 65 public health department grantees to develop and implement a collaborative planning process with their state and local communities as a condition of continued HIV prevention funding. The HIV prevention community planning process offers an unprecedented opportunity for important changes in HIV prevention policy and governmental systems change through local citizen action. We examined the perceptions and experiences of members of community planning groups (CPGs) with respect to systems change and policy making in HIV prevention and identified a series of factors that either promote or inhibit systems and policy change by CPGs. Although there is reportedly substantial support from public health departments for policy making by CPGs, no official guidance supports these activities. CPGs in California have made profound changes in government systems and these experiences position them for policy making in HIV prevention. PMID- 12739795 TI - Science, values and the public health agencies. PMID- 12739796 TI - New record of cluster fly Pollenia rudis (Fabricius) with distribution of all known blow flies (Diptera: Calliphoridae) of Saudi Arabia. AB - During a survey of dipterous flies from Riyadh province, a newly recorded of the cluster fly Pollenia rudis (Fabricius) (Diptera: Calliphoridae) were collected from Saudi Arabia for the first time. A total of 2 males and 3 females were collected from Wadi Haniffa, Riyadh City during the period from February to March 1999 using an aerial net. Data on the morphology, veterinary importance, biology and geographical distribution are presented. Blow-flies lift up to 17 taxa known from the country. A summary of the zoogeographical check up to the 17 species, is presented with widespread distribution for each species. PMID- 12739797 TI - Secretory and humoral antibody responses to Blastocystis hominis in symptomatic and asymptomatic human infections. AB - The study included 3 groups of individuals, in the first 2 groups they had positive stool microscopic examinations only for B. hominis indicating blastocystosis, with and without gastrointestinal symptoms, respectively, while the last group included apparently healthy individuals with no parasites in stool. Stool and serum samples of these individuals were subjected to detection of anti-B. hominis fecal and serum IgA and serum IgG antibodies by indirect ELISA, and detection of B. hominis fecal and serum antigens by double sandwich ELISA. In symptomatic B. hominis infections with positive stool microscopy the study recorded first: specific secretory IgA and humoral IgA and IgG antibody responses at a prevalence of 100%, 83.3% and 86.6%, respectively, with an increased significant difference (P<0.001) of each from healthy controls, together with an increase in level of secretory IgA than that of humoral IgA antibody (P<0.001), and second: the presence of specific antigens in stool and serum at a prevalence of 96.6% and 90%, respectively. With an increased significant difference (P<0.001) of each from healthy controls together with the former at a higher level than the latter (P<0.05). In asymptomatic B. hominis infections with positive stool microscopy the study recorded first; absence of each of the studied specific secretory and humoral antibody responses with no significant difference (P>0.05) of each from healthy controls, and second; absence of specific antigens in stool and serum with no significant difference (P>0.05) of each from healthy controls nor from each other. The explanations and implications of these results are discussed. PMID- 12739798 TI - Ultrastructral studies on the tegument of Dicrocoelium dendriticum (Rudolphi, 1819) Looss 1899. AB - The tegument of Dicrocoelium dendriticum was studied by scanning and transmission electron microscopy. It consists of an anucleate surface syncytium and a deeper nucleated zone. The syncytial layer covers the entire surface of the fluke. It is connected to the nucleated zone through thin cytoplasmic strands. The syncytium is bounded by an apical plasma membrane which is highly folded. This layer mainly contains abundant mitochondria of varying size and shape, and two types of granules, dumbbell shaped and spherical. Although the apical surface shows active exocytosis, no evidence for endocytosis is observed. This is the first study from the State of Kuwait showing the presence of this liver fluke in sheep imported from other countries. PMID- 12739799 TI - Peptone-liver (P-L) a new culture medium for the Egyptian Leishmania major. AB - Leishmania major promastigotes isolated from an Egyptian ZCL patient, was inoculated 4 x 10(6)/ml in a new culture medium, peptone-liver (P-L), with a final concentration of 10% of FCS (fetal calf serum), and incubated at 25+/-1 degrees C. Meanwhile, the same number of promastigotes were inoculated in Schneider's Drosophila medium as control. On the 10th day, number of L. major promastigotes reached 29,25 x 10(6)/ml. The same number was obtained with Schneider's Drosophila medium within four days. The new culture medium or P-L medium is cheap, easily prepared, and gave a very good number of promastigotes as in Schneider's Drosophila medium but in a longer time. PMID- 12739800 TI - Eucalyptus globulus (camphor oil) against the zoonotic scabies, Sarcoptes scabiei. AB - In Egypt, Scarcoptes scabiei is the most prevalence mites of medical importance infesting man. This is true in overcrowded and poorly hygienic areas. Many effective acaridical regimens are available, but being compounds used as insecticides. With the success of camphor oil in treated human demodicidosis, it was applied for treating human scabies. Camphor oil with or without glycerol dilutions gave complete cure, with concentrations (100%, 75% and 50%) within five to ten days. PMID- 12739801 TI - Stigma reduction and improved knowledge and attitudes towards filariasis using a comic book for children. AB - WHO has initiated a global program for lymphatic filariasis (LF) elimination by year 2020. A comic book was designed to improve knowledge and attitudes of Egyptian school children, which included messages on the acceptability of Mass Drug Administration (MDA) and stigma reduction. Comic book administration significantly reduced the fear of the studied children from LF as a killer disease. It helped in positively changing the attitudes of the children towards Elephantiasis patients (p-value <.001). The comic book also reduced the number of children who had earlier stated that they would avoid someone with LF. Knowledge about the ability of treating and preventing LF was also significantly increased among the children after reading the comic book. Moreover, comic book reading helped in raising the awareness towards MDA as the method of choice in preventing LF. Most of the children liked the comic book and its contents. Importantly, 96.2% found this book easy to understand. Many of relatives and friends read the comic book within 2 weeks after distribution. A well-accepted comic book for children is a proven way to reduce stigma and increase knowledge about disease prevention and treatment. PMID- 12739802 TI - Evaluation of an IgG cystatin capture enzyme-linked immunosorbent assay for the detection of anti-cysteine proteinase antibodies in asymptomatic trichomoniasis patients. AB - All isolates of T. vaginalis release cysteine proteinases proteolytic enzymes that are shed into the vagina or culture medium. Cystatin has been used successfully as a capture agent in ELISA to detect cysteine proteinase antibodies without the need for purified proteinases. The ELISA was evaluated in comparison to wet mount microscopy and culture techniques. IgG cystatin capture ELISA proved to be a sensitive and highly specific (100%) assay that could rapidly detect anti cysteine proteinase antibodies in both vaginal washouts and sera of asymptomatic patients with a sensitivity of 100% and 86.7%, respectively. A defined discrimination between sero-positive and sero-negative individuals was markedly observed for ELISA-vaginal washouts providing a more conclusive diagnosis by this technique. The results demonstrated that in trichomoniasis patients (52 cases) whether symptomatic or asymptomatic. T. vaginalis infection was detected in 19 (36.5%), 49 (94.2%), 50 (96.1%) and 48 (92.3%) by wet mount, culture, cystatin capture ELISA-vaginal washouts and ELISA-sera, respectively. The assay was reliable also as a test of cure with a specificity of 94.4% in the vaginal washouts and 83.3% in sera. PMID- 12739803 TI - A preliminary study on dicrocoeliasis in Egypt, with a general review. AB - Dicrocoelium dendriticum (D. dendriticum) is a lancet-shape liver fluke that affects sheep and mammals including man. Human and animal infections have been world widely reported particularly in some of the Eastern Mediterranean sheep raising countries. Infection is acquired by eating the second intermediate host (17 species of ants) with raw fruits, vegetables, herbs or even with the drinking water. As for the first intermediate host, there is about 54 different species of land snails. In spite of the fact that both the first and second intermediate hosts of D. dendriticum are available in Egypt, data about human and animal dicrocoeliasis is lacking. This investigation is a preliminary study to report on the presence of D. dendriticum (in sheep, goats and man in North Sinai Governorate) in Egypt. PMID- 12739804 TI - Effect of itraconazole and terbinafine on Leishmania promastigotes. AB - The antiproliferative effect induced in vitro by two antifungal compounds, the azole itraconazole and the allylamine terbinafine on Leishmania major, L. donovani and L. mexicana promastigotes are reported. Treatment of promastigotes cultures with itraconazole or with terbinafine induced growth arrest with L. major but neither with L. donovani nor with L. mexicana. Concentrations of 0.75 microl/l or more of itraconazole induced cell lysis after 72 hours with L. major. However, even relatively large concentrations of terbinafine (2.0 microl/l) did not induce cell lysis. For L. major, the IC 50 for itraconazole and terbinafine were 0.31 microl/l and 3.3 microl/l respectively. PMID- 12739805 TI - Comparative study of the prophylactic and therapeutic effects of paromomycin, recombinant IL-12 alone or in combination against Cryptosporidium parvum infection in immunosuppressed mice. AB - Administration of paromomycin (100 mg/kg orally) for 10 days, rIL-12 (0.5 ug/mouse s.c.) for 3 consecutive days or combination of both was evaluated before and after infection with C. parvum using immunosuppressed mice model. A total of 110 suckling albino mice were immunosuppressed by hydrocortisone acetate and infected with 10(6) Cryptosporidium oocysts. Assesment of drug efficacy was done by estimating the oocyst count in stool using modified Ziehl Neelsen technique, the histopathological examination of terminal ileum and determination of serum level of IFN-gamma and calculation of the cure rate. The combination of paromomycin and rIL-12 was more effective than either drug alone. The cure rate was 86.7% when the regimen used prophylactically and 73.3% when the combination administered. Regression of the histopathological changes in comparison to the control group was noted. Moreover the combination regimens produced significant higher level of IFN-gamma suggesting that both rIL-12 and Paromomycin can act additively or synergistically in the prevention of C. parvum infection most probably through activation of INF-gamma production. PMID- 12739806 TI - Evaluation of excretory/secretory Fasciola (Fhes) antigen in diagnosis of human fascioliasis. AB - No doubt, human fascioliasis is an increasing worldwide zoonotic liver fluke. Clinically, human fascioliasis has to be differentially diagnosed from many hepatic diseases as acute & chronic hepatitis, schistosomiasis mansoni, visceral toxocariasis, visceral leishmaniasis, hepatic amoebiasis, biliary tract diseases and others. The parasitological diagnosis based on the demonstration of the eggs in stool, duodenal contents or bile is usually unsatisfactory due to false passage of eggs, ectopic fascioliasis, and failure of immature worm to maturation. So, ELISA-Fhes antigen (Fasciola hepatica excretory/secretory) and IHAT were evaluated in the immunodiagnosis of parasitologically proven cases of human fascioliasis compared with proven cases of human schistosomiasis mansoni and parasite-free individuals. ELISA-Fhes gave 100% sensitivity and 100% specificity. On the other hand, IHAT was less sensitive and less specific. PMID- 12739807 TI - Human infection with Dicrocoelium dendriticum in Riyadh district (Saudi Arabia). AB - Dicrocoelium dendriticum (the lancet fluke) is less common liver parasite. Adults inhibit bile duct, while characteristic eggs pass in stools. During one academic year, a total of 1196 patients referred to Parasitology Laboratory from King Abdel Aziz University Hospitals, 121 patients were positive for D. dendriticum. The majority was Saudis, and only 32 of the 121 patients (26.4%) had true dicrocoeliosis, as detected by re-examination after three days of liver free diet. The other patients have false Dicrocoeliasis. The infection was influenced by the patients' behaviours. The Saudis consume more sheep meat during month (Du Elheja) of Haj and Feast. Examination of the slaughtered sheep at the governmental slaughterhouses showed the majority of them have the liver infected (9-24%) with the lancet liver fluke. It is concluded that rate of infection proportionates with the rate of sheep infection and the consumption of infected livers. For proper estimation of human infection should be on diet free liver before examination. PMID- 12739808 TI - Effect of diazinon and/or praziquantel on selected protein aspects in healthy and Schistosoma mansoni infected mice. AB - In Egypt, schistosomiasis is still a major public health problem and praziquantel is the drug of choice for its treatment, whereas diazinon is globally used as an insecticide for controlling pests. They adversely affect the environment. Therefore, the authors studied the effect of 1/20 LD50 diazinon given orally to healthy and Schistosoma mansoni infected mice for 5 successive days up to 9 and 17 weeks coupled with a therapeutic dose (2 x 500 mg/kg Bwt) of praziquantel, 2 weeks before sacrificing. The results showed that non significant differences were obtained from total proteins, albumin, globulins, and albumin/globulin ratio. However, significant differences were revealed from alpha1-, alpha2-, beta1-, beta2-, and gamma-globubins in addition to plasma ceruloplasmin. Diazinon changed the levels of alpha2-, beta1-, and gamma-globubins, while diazinon coupled with schistosomiasis affected the levels of most studied parameters. Consequently, exposure to insecticides should be avoided specially in the rural areas where schistosomiasis is still endemic. PMID- 12739809 TI - Role of endothelial cell adhesion molecules in the development of allergy in human fascioliasis. AB - The recent understanding of the key role of adhesion molecules in the pathogenesis of chest allergy in parasitic infections may provide a pharmacological target for the associated asthmatic symptoms. Circulating levels of the endothelial cell adhesion molecules (CAMs): sICAM-1, sVCAM-1 and sE selectin in sera of 18 allergic asthmatic patients. 16 fascioliasis cases (acute & chronic), 24 fascioliasis cases with allergic chest manifestations and 10 apparently healthy control subjects were estimated by ELISA method. Also, IL-4 serum level was evaluated in all groups. Chest allergy in association with fascioliasis included mainly bronchial asthma, beside eosinophilic bronchitis, persistent wheezing and chronic cough. The study provided evidence that adhesion molecules expression is up regulated in acute and chronic fascioliasis cases with allergic chest manifestations. sVCAM-1 seemed to be an early indicator of asthma development in human fascioliasis. IL-4 cytokine was suggested to be responsible for the increased expression especially in the chronic phase of the disease, yet the role of other cytokines cannot be excluded. PMID- 12739810 TI - Nosocomial sources of cryptosporidial infection in newly admitted patients in Ain Shams University Pediatric Hospital. AB - Cryptosporidium parvum has a world wide distribution. Numerous nosocomial outbreaks of cryptosporidiosis have occurred, also numerous well documented water born outbreaks have occurred. The present study included 104 patients and 27 contacts, both were subjected to stool examination by MZN stain. ELISA for detection of Cryptosporidium Ag in stool was further carried out for the patients developing diarrhea after admission (12), patients admitted with diarrhea (17), and for contacts. Twenty four water samples collected from Pediatric Hospital, and examined by MZN and ELISA. The results showed that 11.5% of examined cases developed diarrhea after admission. 8% of them were positive for cryptosporidiosis by both MZN and ELISA. 17% (3/27) of the contacts were negative by MZN stain. ELISA was found to be 100% sensitive, 94.1% specific. Water samples were found to be negative for C. parvum. PMID- 12739811 TI - Biological and histopathological studies of some plant extracts on larvae of Culex pipiens (Diptera: Culicidae). AB - The water extract of Artemisia monosperma, Eichhornia crassipes and Zygophyllum coccineum was tested against the 2nd instar larvae of Culex pipiens and the resulting pupae and adults. The larvae were exposed to serial concentrations (0.5, 1.0 and 2.0%). All extracts showed marked effects on larval mortality, pupation and adult emergence as compared to the non-treated group. These effects were dose (concentration) dependant (P < 0.001). The results indicate that the water extract of E. crassipes gave marked and significant effects, followed by A. monosperma and Z. coccineum particularly at 2% concentration (P < 0.001). The water extract of E. crassipes at 2% conc. revealed drastic effect on larval midgut, integument, fats and muscles. These histopathological effects increased with increasing the time of exposure (treatment). PMID- 12739812 TI - A longitudinal study of schistosome intermediate host snail populations and their trematode infection in certain areas of Egypt. AB - Seasonal variation of Biomphalaria alexandrina and Bulinus truncatus populations and their infection rates with schistosome and other trematode cercariae were studied longitudinally in four water courses located in Giza and Faiyoum Governorates. Abundance of both species varied from year to year and according to the type of habitat. The mean prevalence of Schistosoma mansoni in Biomphalaria was 0.29%, that of S. haematobium in Bulinus was 1.36%. Seasonal variations of age structure of the 2 vector snails were monitored throughout the survey period. Infection rates with schistosome and other trematodes among Bulinus and Biomphalaria increased with the increase in snail size. Data suggest the occurrence of an antagonistic interaction between schistosome and non-human cercariae, especially echinostome, in infected snails. PMID- 12739813 TI - Vaccination trial against experimental trichrinellosis using autoclaved Trichinella spiralis larvae vaccine (ATSLV). AB - The autoclaved Trichinella spiralis larvae vaccine (ATSLV) was tested and showed a surprising and somewhat unpredictable effect on the immune system of mice experimentally infected with T. spiralis. The vaccine was given with Bacille Calmette Guerin (BCG) as an adjuvant at different durations and by different routes of administration. The best result was achieved by given the vaccine twice intradermally with two weeks interval, as evidenced by a significant reduction in adult and larval count, as well as reproductive capacity index. Histopathologically, there was significant reduction in number of the encysted larvae which showed degeneration and hyalinization of the cyst wall accompanied by early pericystic fibrosis. PMID- 12739814 TI - Electron miscroscopy and histochemical studies on four Egyptian helminthes eggs of medical importance. AB - By SEM the Fasciola gigantica egg is ovoid with a small knob like operculum, while the egg of Heterophyes heterophytes is broad oval with the operculum more tapering. The egg shell of fertilized Ascaris lumbricoides has interconnected ridges and peak-like projections, while the egg of Enterobius vermicularis is flattened with a thicker margin at the curved side. By TEM, Fasciola egg shell consists of fine reticulum fibrils of three layers. The outer lipoprotein of perivitelline membrane beneath which 2 membranes separated by inclusions, middle of protein globules and inner lipoprotein layer with minute electron-dense granules of melanin or polymer origin, in some parts of the shell giving the egg its brown coloration. The Heterophyes egg shell is more or less similar to that of Fasciola but lacking the minute electron-dense granules. The egg shell of Ascaris has outer ulterine layer with three consecutive layers, basal lipoprotein layer and the inner lipid or ascaroside layer which is the most resistant layer. The Enterobius egg shell consists of five layers, external uterine, internal uterine, vitelline, chitinous and lipid layer. Histochemically, Fasciola egg shell consists of nine amino-acids, and that of Heterophyes consists of ten amino acids. In Ascaris, the lipid layer characteristically consists 25% protein and 75% lipid. The histochemical examination of Enterobius as a detailed example, showed different degrees of reactions with mercuric bromophenol blue, diazotization coupling, Sakaguchi reaction, Sudan black and Mallory's triple stain. Temperature showed marked effect on eggs survival. Eggs of Fasciola and Heterophyes withstand more low temperatures but those of Ascaris and Enterobius withstand more high ones. There are marked correlations between the egg shell constitution, histochemical compositions on one hand and water permeability and egg dryness on the other hand. The results were photographed and discussed. PMID- 12739815 TI - Biochemical changes after subchronic and chronic interaction of Schistosoma mansoni infection in Swiss albino mice with two specific compounds. AB - In Egypt, infection with Schistosoma mansoni (S.m.) and residues of pesticides have been considered as major environmental pollutants that adversely affect health. Effects of diazinon (DZN) and/or praziquantel (PZQ) on the levels of plasma triiodothyronine (T3), thyroxine (T4), activities of brain acetylcholinesterase (AchE) and liver alanine aminotransferase (ALT) in addition to blood reduced glutathione (GSH) in healthy and S.m. infected mice were investigated after 9 and 17 weeks of either infection or intoxication with DZN. Triiodothyronine showed significant differences among the different treatments. The group of mice treated with PZQ showed the highest levels of T3 at both time intervals. Thyroxine level showed significant differences between the two time intervals. The lowest levels of T4 were observed in the infected-PZQ group at week 17. The maximum inhibition of brain AchE activity was noticed in DZN-PZQ treated group after 9 and 17 weeks. The different treatments significantly reduced the activities of liver ALT. The highest decrease was recorded in the infected-DZN-PZQ group at week 9. All treatments significantly lowered the levels of blood GSH after 9 weeks. PMID- 12739816 TI - Genotypic discrimination between a Leishmania isolate and two Leishmania major reference strains using PCR-RFLP technique. AB - The restriction fragment length polymorphism (PCR-RFLP) of 18S rDNA amplified fragments, was conducted to recognize between an unidentified Leishmania isolated from Egyptian patient infected in Saudi Arabia and two L. major reference strains causing cutaneous lesions. The strains were maintained both in vivo & in vitro. Additional requirement as morphological characterization on basis of the light microscope & scanning electron microscopy and behaviour in experimental Swiss albino mice regarding development of lesions were performed. The results showed that, PCR-RFLP analysis of the 18S rDNA amplified PCR fragments was highly successful to put the unidentified Leishmania strain in the category of L. major. There was no significant differences regarding the cutaneous lesions development. In spite of the significant variations of the morphometric measurements of the three strains were observed. PMID- 12739817 TI - Cystatin capture-dot-enzyme-linked immunosorbent assay for immunodiagnosis and assessment of cure of experimental trichinellosis in mice. AB - Cystatin capture dot enzyme-linked immunosorbent assay (CC-dot ELISA) was evaluated as a new version of ELISA for diagnosis of trichinellosis by the detection of anti-Trichinella spiralis cysteine proteinase (CyP) IgG, using nitrocellulose membrane sensitized with cystatin as a capture reagent for CyP from T. spiralis muscle larvae excretory secretory products (ESP) without purification, compared to the detection of anti-T. spiralis IgG by conventional (conv.) ELISA, using whole ESP. Experimentally infected mice with light (100 larvae/mouse) and heavy (300 larvae/mouse) infections by T. spiralis larvae at 7, 14, 21 and 56 days post infection, and after flubendazole treatment were examined. As early as one week post infection CC-dot ELISA gave high positive rates of 86.6% and 100% in light and heavy infections, respectively, in contrast to negative results by conv. ELISA. CC-dot ELISA showed in light and heavy trichinellosis a higher efficiency in comparison to conv. ELISA (97.7% versus 67.7% and 98.8% versus 80%, respectively) and a higher overall sensitivity (96.6% versus 55% and 98.3% versus 73.3%, respectively). No cross reactions with sera of other parasitic infected or non infected control mice were recorded by CC-dot ELISA giving 100% specificity compared to 93.3% by conv. ELISA. After treatment, CC-dot ELISA gave positive results only in uncured mice with remaining muscle larvae, while conv. ELISA was positive in mice with and without remaining muscle larvae. CC-dot ELISA used lower quantities of antigen, was performed at room temperature, and read by naked eye in less than 2.5 hours. PMID- 12739818 TI - Preliminary field trials with Culicinomyces clavosporus against some Egyptian mosquitoes in selected habitats. AB - Culicinomyces clavosporus was isolated from diseased larvae of Culex pipiens collected from brackish water habitat. Pure cultures of the fungus were grown on nutrient agar media. In the laboratory, the Egyptian isolate of the fungus was bioassayed against first instar larvae of Aedes caspius, Cx. antennatus and Cx. pipiens. The estimated LC50's for Ae. caspius, Cx. antennatus and Cx. pipiens were 4.4 x 10(2), 7.6 x 10(2) and 11.8 x 10(2) conidia/ml, respectively. Field studies were undertaken in a variety of habitats supporting populations of mosquitoes in Mahalet Marhoom, El-Gharbia Governorate, in the summer of 1999. Evaluations included artificial pool studies and field trials. The fungus was applied once to 3 different natural breeding habitats of mosquitoes at a dose rate of 10(10) conidia/m2. Cx. clavosporus introduction into unpolluted rice field that had high densities of Cx. antennatus and low densities of Anopheles tenebrosus Doenitz and Cx. perexiguus resulted in 100% control of the larvae 5 days post treatment. Introduction of the fungus into brackish water habitat supporting large brood of Ae. caspius, and few numbers of Culiseta longiareolata (Macquart), Cx. Pusillus, Cx. perexiguus and Cx. pipiens was equally effective. Dissection and examinations of larvae from both sites confirmed infection by the fungus. However, introduction of the fungus into a drain supporting high population density of Cx. pipiens was not effective. Microscopic examination of larvae removed from this site revealed that the conidia failed to germinate and penetrate the host cuticle. The presence of organic pollution in the breeding site drastically reduced the infectivity of the fungus. Thus, the fungus appears promising as a possible limiting factor for mosquito populations, and eventually its use in control of mosquitoes merits further investigations. PMID- 12739819 TI - The effect of potassium permanganate and sodium dichloroisocyanurate on metacercariae of Fasciola gigantica. AB - Batches of encysted metacercariae of F. gigantica, adhered to transparent polyethylene sheets, were treated with KMnO4, while others were treated with NaDCC at specific concentrations and exposure times. Assessment of the effects was carried out by the detached percentage and viable metacercariae and by scanning electron microscope (SEM) ultrastructure changes. In addition, their effects on leaves of green vegetables were reported. The results showed that all metacercariae were detached and were dead by exposure to KMnO4 (96%) and NaDCC (100%) were detached from the polyethylene sheets. SEM showed that the deformities in the metacercariae soaked in NaDCC were more severe than those dipped in KMnO4. However, neither KMnO4 nor NaDCC affected the consistency, color, taste or flavor of the vegetables' leaves. The two disinfectants particularly NaDCC, proved to be safe, effective against the encysted metacercariae with no side-effects. PMID- 12739821 TI - Ultrastructural and immunological features of experimental cutaneous leishmaniasis after treatment with intralesional hypertonic sodium chloride and CO2 laser rays. AB - This study investigated the mechanism of action and efficacy of CO2 laser rays and hypertonic sodium chloride (NaCl) with different concentrations in treatment of cutaneous leishmaniasis (CL) as assessed clinically, parasitologically, histopathologically by light & transmission electron microscopy (TEM) and immunologically by RT-PCR for gene expression of interleukin-13 (IL-13). Eighty mice were divided into four groups. The first was non infected control group (n=16) and the second was infected and served as non-treated control (n=16). The third (n=32) and fourth groups (n=16) were subjected to NaCl injection and CO2 laser respectively. Results showed that clinical healing by CO2 laser were nearly similar to normal appearance, but differed according to concentration of NaCl, as confirmed by ultrastructure and immunohistopathologic features of the host cells and surrounding skin tissue. IL-13 mRNA was significantly decreased after treatment denoting that Th2 cytokine (IL-13) is important for the development of strategies to prevent the induction of pathologic processes. It is concluded that CO2 laser then 7% NaCl are good modalities for CL treatment and is recommended wherever possible. PMID- 12739820 TI - Demonstration of species-specific and cross-reactive components of hydatid cyst fluid antigens. AB - ELISA-hydatid cyst fluid antigen (HCF-Ag-ELISA) and conventional echinococcosis IHAT were assessed for cystic hydatid disease (CHD), confirmed by HCF-Ag immunoblotting assay. The sensitivity of the tests was 94.4% and 83.3% for HCF-Ag ELISA and IHA respectively. HCF-Ag-ELISA showed some cross-reaction in tumor cases (5.5%), but no cross-reaction was observed in Rheumatoid arthritis patients. IHA showed 100% specificity. To confirm this data SDS-PAGE immunoblotting using HCF-Ag was applied. Echinococcus specific reactions were reported with 44Kda, 34Kda, 29Kda and 8Kda HCF-Ag resolved bands. Cross-reaction was found with 27Kda, 21Kda, 16Kda and 13Kda bands in tumor patients. Moreover, cross-reaction was found with 200Kda, 175Kda, 62Kda, 52Kda and 40Kda in serological anti-Schistosoma antibodies positive cases. So, using specific HCF-Ag components, immunoblotting provided 100% sensitivity and specificity in CHD. PMID- 12739823 TI - Effect of U-74500A, a 21-aminosteroid on renal ischemia-reperfusion injury in rats. AB - Renal ischemia-reperfusion injury constitutes the most common pathogenic factor for acute renal failure and is the main contributor to renal dysfunction in allograft recipients and revascularization surgeries. Many studies have demonstrated that reactive oxygen species play an important role in ischemic acute renal failure. The aim of the present study was to investigate the effects of the synthetic antioxidant U-74500A, a 21-aminosteroid in a rat model of renal ischemia-reperfusion injury. Renal ischemia-reperfusion was induced by clamping unilateral renal artery for 45 min followed by 24 h of reperfusion. Two doses of U-74500A (4.0 mg/kg, i.v.) were administered 45 min prior to renal artery occlusion and then 15 min prior to reperfusion. Tissue lipid peroxidation was measured as thiobarbituric acid reacting substances (TBARS) in kidney homogenates. Renal function was assessed by estimating serum creatinine, blood urea nitrogen (BUN), creatinine and urea clearance. Renal morphological alterations were assessed by histopathological examination of hematoxylin-eosin stained sections of the kidneys. Ischemia-reperfusion produced elevated levels of TBARS and deteriorated the renal function as assessed by increased serum creatinine, BUN and decreased creatinine and urea clearance as compared to sham operated rats. The ischemic kidneys of rats showed severe hyaline casts, epithelial swelling, proteinaceous debris, tubular necrosis, medullary congestion and hemorrhage. U-74500A markedly attenuated elevated levels of TBARS as well as morphological changes, but did not improve renal dysfunction in rats subjected to renal ischemia-reperfusion. These results clearly demonstrate the in vivo antioxidant effect of U-74500A, a 21-aminosteroid in attenuating renal ischemia reperfusion injury. PMID- 12739822 TI - Role of quercetin on hepatic urea production in acute renal failure. AB - Acute renal failure (ARF) is a serious damage of renal function induced by various nephrotoxic drugs, ischemia, bilateral urethral obstruction, trauma and unilateral nephrectomy. Dramatic clinical syndrome, azotemia, develops as a result of hypovolemia, oliguria, reduced glomerular filtration and acidosis. In addition to classic medications recent studies give more attention to beneficial effect of natural plant products as bioflavonoids. We have studied the influence of bioflavonoid, quercetin, on hepatic urea production in glycerol induced ARF in the rats. Male Sprague Dawley rats were used in the experiment. The value of urea production in the liver was determined by measuring of liver arginase activity, the terminal enzyme of urea cycle. Arginase activity was increased (p < 0.01) as well as urea level (p < 0.001) 48 h after glycerol administration. Pretreatment by quercetin suppressed the arginase activity in the liver (p < 0.05) and plasma levels of urea (p < 0.01). So, we have concluded that quercetin may be beneficial in glycerol induced ARF. PMID- 12739824 TI - Vesicoureteric reflux and reflux nephropathy as seen at a tertiary care adult nephrology service in India--an analysis of 86 patients. AB - Clinical features and risk factors for renal failure in patients with reflux nephropathy (RN) as seen in an adult nephrology service are likely to be different than those seen in a pediatric service. There are only a few studies on adults with vesicoureteric reflux (VUR) and RN and data on RN as seen in developing countries is still evolving. Retrospective analysis of records of patients diagnosed to have VUR by conventional micturating cystourethrogram over a 13 year period, as seen in the adult nephrology services of this tertiary care hospital in north India was carried out. Results are presented as mean +/- 2 SD. Unpaired t-test was used to compare means, chi-square test to define associations, and logistic regression analysis was done to define risk factors. Out of 86 patients diagnosed to have VUR, 69 (80.2%) were males and 22 (25.6%) were children. The mean age at presentation was 24.3 +/- 14.5 years and at onset of symptoms was 19.64 +/- 14.8 years. Sixty-nine (80.2%) patients had chronic renal failure (CRF) at presentation, including 33 (38.4%) patients who already had end stage renal failure (ESRF) at presentation in whom reflux was diagnosed during routine pretransplant evaluation and these constituted 5.5% of all ESRF patients. The clinical features at presentation were hypertension in 51 (59.3%), recurrent urinary tract infection (UTI) in 31 (36.1%), history of stones in 7 (8.1%), and gross hematuria in 4 (4.7%). Patients with history of recurrent UTI were more likely to be females (p < 0.01) and to present without renal failure (p < 0.05). Proteinuria > 1 g/day was significantly associated (p < 0.02) with hypertension at presentation. Patients who presented with renal failure were more likely to be males (p < 0.05), not to have history of recurrent UTI (p < 0.05), have proteinuria > 1 g/day (p < 0.02) and higher grades (grades IV and V) of reflux (p < 0.05). On logistic regression analysis, higher age of onset (odds ratio 4.6, p < 0.03), proteinuria > 1 g/day (odds ratio 3.8, p < 0.05), and male gender (odds ratio 3.5, p < 0.05) were significant risk factors for presentation for the first time with renal failure. The clinical features and course of VUR and RN as seen in India are different from those reported from elsewhere. The vast majority of patients in India are males and almost two thirds do not have a past history of UTI. Renal failure is present in more than three fourths of patients when a diagnosis of reflux is made and one third of all patients present with ESRD. Patients with a prior history of UTI are more commonly females and are less likely to have renal failure at presentation. Higher age of onset of symptoms, proteinuria > 1 g/day and male gender were risk factors for the development of renal failure. It is likely that these asymptomatic patients remain undetected during childhood, presenting late only after having incurred severe renal damage. PMID- 12739825 TI - Combination of continuous renal replacement therapies (CRRT) and extracorporeal membrane oxygenation (ECMO) for advanced cardiac patients. AB - BACKGROUND: The critically ill patients may require mechanical ventilation, cardiac mechanical support, and other types of critical support. Extracorporeal membrane oxygenation (ECMO) is a supportive therapy, which provides good cardiopulmonary and end-organ support. Continuous renal replacement therapies (CRRT) exhibit important advantages in terms of clinical tolerance and blood purification. This investigation aims to evaluate the acute renal failure in cardiac patients under ECMO, and assess the effect of combining these two technologies, ECMO and CRRT. METHODS: Between December 1998 and June 2001, 10 adult cardiac patients were treated on ECMO. Five of them were treated with both ECMO and CRRT. The clinical outcomes were retrospectively analyzed. RESULTS: Of the 10 patients studied, five were men and five were women. The mean age of survivors and non-survivors was 37.00 +/- 14.54 years and 46.17 +/- 7.41 years, respectively. The overall mortality rate was 60%. Survivors did not differ significantly from non-survivors in age or gender. The APACHE II scores on the first day of ECMO support between survival and non-survival were 19.00 +/- 9.38 and 24.67 +/- 3.50 (P value = 0.392) (Table 2), which demonstrates no significant differences too. The cause of death in most patients was related to organ system failure during the 24 h immediately before ECMO started. Five patients with acute renal failure treated by CRRT were eventually died. The median and mean survival in this group on CRRT was 40.50 +/- 18.07 h and 92.60 +/- 60.50 h. CONCLUSION: We conclude that mortality rate for acute renal failure in cardiac patients under ECMO continues to be high. Our data suggest that acute renal failure is generally a part of multiorgan failure. This unique form of acute renal failure, causes generalized edema and fluid overload despite still low serum creatinine and azotemia, and deteriorates rapidly to death. From this study shows, advanced cardiac failure may need more aggressive and early initiation of ECMO support before acute renal failure develops. Acute renal failure in advanced heart failure under ECMO support means a grave sign, need aggressive heart transplantation therapy as soon as possible. Combination of CRRT and ECMO might serve an alternative therapy bridging the temporary replacement treatment and heart transplantation in advanced cardiac patients. PMID- 12739827 TI - Hyperhomocysteinemia and oxidative stress during dialysis treatment. AB - BACKGROUND/AIMS: The concomitant presence of hyperhomocysteinemia and oxidative stress may represent a determinant factor for the occurrence of vascular alterations and cardiac diseases, the main cause of death among dialysis patients. The aim was to analyze the occurrence of hyperhomocysteinemia and oxidative stress and their possible relationship in dialysis patients. METHODS: Antioxidant substances, homocysteine, folate, and vitamin B12 were determined in blood from 32 patients on hemodialysis (HD), 21 patients on peritoneal dialysis (PD), and 12 healthy individuals. RESULTS: Different degrees of hyperhomocysteinemia were observed in all HD patients and in 95% of the PD patients (45.30 +/- 24.89 microM in HD and 35.50 +/- 26.53 microM in PD). Oxidative stress defined as an imbalance between oxidant and antioxidant forces was observed in all dialysis patients, but was more intense in HD individuals. In this group, lipoperoxidation and protein oxidation were associated with lower concentrations of antioxidants such as erythrocyte vitamin E and vitamin C. CONCLUSIONS: Hyperhomocysteinemia and oxidative stress occur in both types of dialysis treatment, possibly contributing to the establishment of complications in these patients. PMID- 12739826 TI - Plasma homocysteine levels in Indian patients with essential hypertension and their siblings. AB - BACKGROUND: An association between plasma homocysteine and essential hypertension and their non-hypertensive siblings is controversial. METHODS AND RESULTS: Plasma homocysteine levels were measured in subjects with essential hypertension (n = 50), their normotensive siblings (n = 50) and normotensive controls (n = 50). All the subjects were non-diabetic, had no past history of myocardial infarction, stroke or peripheral vascular disease and had normal renal functions. The mean homocysteine values were 18.96 +/- 8.08 micromol/L in patients, 14.84 +/- 5.55 micromol/L in siblings and 10.50 +/- 4.92 micromol/L in controls. Plasma homocysteine level were found to be significantly higher in patients with hypertension (p < 0.0001) and their normotensive siblings (p < 0.0001) when compared to controls. Also patients with hypertension had their higher plasma homocysteine levels compared to their siblings (p < 0.0036). CONCLUSION: Plasma homocysteine levels are significantly elevated in Indian patients with essential hypertension and their normotensive siblings. Thus, plasma homocysteine may serve as a marker for the development of essential hypertension. PMID- 12739828 TI - Factors influencing the non-recovery of renal function after the relief of urinary tract obstruction in women with cancer of cervix. AB - OBJECTIVES: To identify factors associated with non-recovery of renal function after the relief of urinary tract obstruction in women with cancer of cervix. PATIENTS AND METHODS: Thirty-seven women with cancer of cervix and obstructive renal failure were prospectively studied. Patients were evaluated in the day before nephrostomy (D0); and in the 1st (D1), 7th (D7), and 30th day (D30) after unilateral nephrostomy. The following parameters were analyzed: age serum urea, and serum creatinine (S(creat)), in D0, D7, and D30. Renal cortical thickness was measured by ultrasonography in D0. During nephrostomy procedure, urine samples for urinalysis and culture were collected and renal biopsy was attempted. Patients were divided in two groups according to S(creat) in D30: patients with S(creat) < 1, 4mg/dL (R group) and those with S(creat) > or = 1, 4mg/dL (nR group). RESULTS: Ten patients died before D30 and were not studied. R group (n = 12) was younger (43 +/- 9 vs. 52 +/- 10 years, p < 0.05) than nR group (n = 15), and presented lower serum urea in D0 (134 +/- 67 vs. 212 +/- 51 mg/dL, p < 0.01) and in D7 (94 +/- 20 vs. 155 +/- 18 mg/dL, p < 0.05). Renal cortex was thicker in R group (17 +/- 1 vs. 13 +/- 1 mm, p < 0.05). Survival was higher in R group (11/12) than in nR group (0/15, p < 0.01). All urine samples showed leukocyturia but culture was positive in only two. In three out of nine biopsy attempts enough material was obtained. Light microscopy revealed interstitial fibrosis plus tubular atrophy, and partial glomerular fibrosis was present in two biopsies. CONCLUSIONS: The non-recovery of renal function after the relief of obstruction in human obstructive renal failure is associated with older age and decreased renal cortical thickness. The complete renal recovery improves patient survival despite malignancy. PMID- 12739829 TI - Acute renal failure due to intrinsic renal diseases: review of 1122 cases. AB - In this study we have analyzed incidence, causes and clinical course of ARF due to primary intrarenal disease other than acute tubular necrosis. Thousand hundred and twenty two cases of ARF of diverse etiology were studied over a period of 16 years; July 1984 to Dec, 1999. Surgical ARF 231 (20.6%) were not included in the present study. Intrinsic renal diseases were responsible for ARF in 891 (79.4%) of cases. The most common intrinsic renal diseases 705 (79.4%) causing ARF were ischemic/toxic acute tubular necrosis, but not included in this study. Acute renal failure was related to acute glomerulonephritis (9.3%), acute interstitial nephritis (7%), and renal cortical necrosis in (4.6%) of cases. Therefore intrinsic renal diseases other than ATN were the causative factor for acute renal failure in 186 (20.8%) patients in our study. Crescentic (51.8%) and endocapillary proliferative glomerulonephritis (34.9%), were the main glomerular diseases responsible for ARF and 75.9% of GN was related to infectious etiology. Fifty three percent of acute interstitial nephritis was drug induced and in 25 (40%) patients it was related to an infectious etiology. Renal cortical necrosis due to HUS was observed in 16 (39%) children and majority (76.47%) of the cases had a diarrhoeal prodrome. Obstetrical complications were the main causes (61%) of cortical necrosis in adults with acute renal failure. Thus, intrinsic renal diseases other than ATN were responsible for ARF in 186 (20.8%) cases. Post infectious glomerulonephritis, acute interstitial nephritis and renal cortical necrosis (complicating HUS in children and obstetrical complications in adult) are the main causes of acute renal failure in our study. Both acute GN and interstitial nephritis had excellent prognosis, however renal cortical necrosis was associated with a very high mortality. PMID- 12739831 TI - Atherosclerosis in patients with end-stage renal failure prior to initiation of hemodialysis. AB - BACKGROUND: In dialysis patients cardiovascular mortality is 10 to 20 times higher than in general population. It remains uncertain whether atherosclerosis of dialysis patients is effectively accelerated because many of dialysis patients have more or less marked vascular lesions already at the start of dialysis treatment. SUBJECTS AND METHODS. Using B-mode ultrasonography (ATL HDI 3000), we compared intima-media thickness (IMT) and plaque occurrence (indicators of atherosclerosis) in the common carotid arteries (CC), in the area of bifurcation (CB) and in the proximal part of internal carotid arteries (CI) in 28 hemodialysis patients (14 men and 14 women; mean age 49.4 years; mean duration of HD treatment 66.6 months) with that in 28 age-sex matched patients prior to initiation of hemodialysis. We also investigated possible differences in atherosclerotic risk factors in both groups. RESULTS: The IMT values of CC (0.71 vs. 0.70 mm; p = 0.937), CB (0.81 vs. 0.77 mm; p = 0,423) and CI (0.72 vs. 0.71 mm; p = 0.935) were not significantly different in dialysis patients and patients starting dialysis treatment. We also found no difference in plaque occurrence (61% vs. 54%; p = 0.787) and in atherosclerotic risk factors (hypertension, smoking, lipids) between both groups. CONCLUSIONS: In our study we found no difference in atherosclerotic lesions in carotid arteries between dialysis patients and patients with end-stage renal failure starting dialysis treatment. Patients with chronic renal failure are at high risk for cardiovascular diseases so we should intervene earlier and more actively long before dialysis treatment in order to reduce the atherosclerotic risk factors. PMID- 12739830 TI - Assessment of protein nitrogen appearance in Chinese peritoneal dialysis patients which method to use? AB - OBJECTIVE: We compared the Bergstrom's and Randerson's formula for PNA determination, and compared the normalization of PNA by ideal body weight (IBW) and standard body weight (SBW) as estimated by the Watson's formula. METHODS. We studied 208 Chinese PD patients. Two 24-h dialysate and urine collections were performed six months apart. Protein nitrogen appearance was determined by the Randerson's formula (PNA-Rand) and Bergstrom's formula (PNA-Berg), the latter used as the gold standard. PNA-Berg was normalized with IBW and SBW, denoted as NPNA-IBW and NPNA-Watson respectively. The change of PNA over six months, denoted as APNA-Rand and APNA-Berg, were calculated. The results were compared by the Bland and Altman's method. RESULTS: At zero month, the average PNA-Berg was 61.8 +/- 14.8 g/day, and the average PNA-Rand was 58.1 +/- 14.5 g/day. The value of PNA-Rand was consistently lower than the corresponding PNA-Berg. The bias of PNA Rand was -3.7g/day. The limits of agreement were -9.2 to +1.8 g/day. When NPNA Watson was compared to NPNA-IBW, the bias of NPNA-Watson, using NPNA-IBW as gold standard, was 0.01 g/kg/day; the limits of agreement were -0.22 to +0.23 g/kg/day. The difference between NPNA-Watson and NPNA-IBW correlated with the body mass index (r = -0.820, p < 0.001) and body weight (r = -0.834, r < 0.001). After six month, there was a significant reduction in urine protein loss. However, total protein loss was only slightly reduced (7.3 +/- 3.0 to 6.9 +/- 2.8 g/day, p = 0.029). The correlation between APNA-Berg and APNA-Rand remained excellent (r = 0.983, p < 0.001). The bias of APNA-Rand was +0.3 g/day; the limits of agreement were -4.7 to +5.2 g/day. CONCLUSION: Ideal body weight that is validated for specific ethnic group, rather than the Watson's formula, should be used for normalization of PNA. Although the Randerson's formula under estimates PNA when compared to the Bergstrom's formula, it is a reliable method for serial PNA monitoring because dialysate protein loss is stable in most patients. PMID- 12739832 TI - Efficacy of GM-CSF as an adjuvant to hepatitis B vaccination in patients with chronic renal failure--results of a prospective, randomized trial. AB - BACKGROUND: Chronic renal failure patients on hemodialysis are at an increased risk of acquiring hepatitis B infection. Hence vaccination against hepatitis B assumes great importance in these patients. However, the response to hepatitis B vaccination is poor, even when 4 double doses (40 microg) of the vaccine are given. This study was conducted to determine the efficacy of GM-CSF as an adjuvant to hepatitis B vaccine in CRF patients. METHODS: CRF patients including both hemodialysis (HD) and non-dialysis (ND) patients were randomized to receive either placebo or a single injection of GM-CSF (in varying doses of 50 microg, 100 microg, 150 microg) a day prior to the 1st dose of recombinant hepatitis B vaccine (40 microg). Three more doses of the vaccine were given at 1, 2, and 6 months. The anti-HBs antibody titres were measured by ELISA at 3 and 7 months. Patients having antibody titres less than 10 IU/L were considered non-responders. The response rate and mean antibody titers were compared between the control (I) and GM-CSF (II) groups. RESULTS: In group I, 31 and 27 patients were available for evaluation at 3 and 7 months respectively. In group II, 33 and 28 patients could be evaluated at the same time points. Within the control group (group I), the response rate in hemodialysis patients (63.6%) was lower as compared to non dialysis patients (81.2%). The response rate in group II was higher than that in group I at both 3 months as well as 7 months (78.1% vs. 62.3% and 89.3% vs. 74.1%, p = ns). The best response rates in group II were observed when GM-CSF was used in a dose of 150 microg (90.9% at 3 months and 100% at 7 months). The mean antibody titers were also found to be higher in the group II as compared to group I (409.6 vs. 243.9 IU/L, p = 0.01). CONCLUSION: The results of this randomized, prospective study suggest that: 1. Patients with chronic renal failure should be vaccinated for hepatitis B as chronic renal insufficiency is established. 2. GM CSF is an effective adjuvant to hepatitis B vaccine in these patients especially when a priming dose of 150 microg is used prior to 1st dose of hepatitis B vaccination. PMID- 12739833 TI - The effects of acute renal failure on long-term renal function. AB - The relationship between acute renal failure (ARF) and long-term renal function remains unknown. We therefore undertook a study of patients at the Baltimore VA Hospital to examine the effects of a bout of acute renal injury on long-term renal function. We retrospectively reviewed the relationship between serum creatinine and time of observation for 6058 individuals who had values greater than 1.4 mg/dL in any two consecutive years. Individuals were stratified according to total years of observation with a minimum of two years. Severity of acute renal injury was divided into mild, moderate, and severe with elevations in baseline creatinine of < 50%, 50-300% and > 300% respectively. Sporadic elevations in creatinine were evident in 8-15% of the population. There were a total of 1328 episodes of acute renal failure in 916 patients that were suitable for analysis. Mild ARF on a substrate of normal or mildly abnormal renal function resolved without long-term sequelae. Moderate and severe ARF occurred more frequently on a background of reduced renal function but baseline function was retained in at least 60% of patients. We conclude that ARF is more frequent in patients with chronic kidney disease but it is not invariably associated with an accelerated course to end-stage renal disease or death. Overall, the majority of ARF events resolved without adverse long-term effects suggesting appropriate management in the majority of instances. PMID- 12739835 TI - LV mass index significantly impacts on patient and renal outcomes in patients with coronary artery bypass grafting and poor left-ventricular function. AB - BACKGROUND: A recent report demonstrated that the presence of left ventricular hypertrophy was an independent predictor of mortality in patients with coronary artery bypass grafting (CABG) severely depressed left ventricular function. However, the impact of left ventricle (LV) mass index on the renal and patient outcomes in such patients with CABG has previously not been addressed. The present study thus considers this group of patient and uses LV mass index to assess renal and patient outcomes for these patients. MATERIAL AND METHOD: All patients who arrived at the emergency room with severe cardiac dysfunction (EF < 60%), triple vessel disease, and required CABG and LV hypertrophy (LVH) (LV mass index gamma110 g/m2 in women, gamma134 g/m2 in men) were admitted preoperatively to the intensive care unit (ICU) for supportive intervention from January 1, 1998 to January 1, 2001. Of all LVH patients, 44 underwent CABG, and were divided into two groups according to LV mass index. RESULTS: Of all patients, 72.7% had severe echocardiographic LVH. The echocardiographic data of both dialysis and non dialysis groups showed no difference with respect to echocardiographic findings. Histories of myocardial infarction were more frequent in the severe LVH group that in the mild LVH group. As for pre-operative systolic blood pressure and diastolic blood pressure, mean systolic and diastolic blood pressure values were significantly lower in the severe LVH group. Ejection fraction was also significantly lower in the severe LVH group than in the mild LVH group. The patients in the severe LVH group were significantly more likely to have received hemodialysis following CABG surgery (62.5% vs. 33.4%, p < 0.05). Mortality was higher in the higher LV mass index group that in the lower LV mass index group (56.2% vs. 25%, p < 0.05). CONCLUSION; Patients with a significantly higher LV mass index usually manifest lower pre-operative blood pressure and poor cardiac function. Consequently, these patients will have a poor renal outcome and higher mortality. PMID- 12739834 TI - Advanced glycation end products in hemodialyzed patients with diabetes mellitus correlate with leptin and leptin/body fat ratio. AB - Advanced glycation end products (AGEs) and other carbonyl and oxidative stress compounds are supposed to play a critical role in the pathogenesis of several diseases and their complications, i.e., diabetes mellitus, diabetic retinopathy, atherosclerosis, and chronic renal failure. In the present investigation, we were interested in the relationship of AGEs in plasma to other prominent factors in the patients on chronic hemodialysis treatment-27 patients with diabetes mellitus, 35 patients without diabetes mellitus. AGE-group reactivity was estimated using a spectrofluorometric method (excitation 350 nm, emission 430 nm) and is expressed in arbitrary units (AU). We found significantly higher AGEs levels in diabetics than in non-diabetics on regular hemodialysis treatment both before (2.7 +/- 0.7 x 10(4) AU vs. 2.2 +/- 0.6 x 10(4) AU, p < 0.001) and after the dialysis session (2.3 +/- 0.5 x 10(4) AU vs. 1.8 +/- 0.7 x 10(4) AU, p < 0.005). AGEs were significantly reduced during hemodialysis in both groups of patients--by 15.4% in the diabetic go (p < 0.001) and by 17.3% in non-diabetics (p < 0.005). In the patients with diabetes mellitus, AGEs did not correlate with parameters of the glucose metabolism correction (blood glucose, HbA1c). We observed a significant correlation between AGEs and leptin (r = 0.48, p < 0.05) as well as the leptin/body fat ratio (r = 0.56, p < 0.05) only in hemodialyzed patients with diabetes mellitus. These findings suggest more detailed studies to identify the molecular links between carbonyl stress, i.e., advanced glycation end products, and leptin metabolism, sign of microinflammation and hypertension. PMID- 12739836 TI - Baclofen neurotoxicity in uremic patients: is continuous ambulatory peritoneal dialysis less effective than intermittent hemodialysis? AB - OBJECTIVES: To compare the efficacy between continuous ambulatory peritoneal dialysis (CAPD) and intermittent hemodialysis in the treatment of baclofen associated neurotoxicity. METHODS: Three uremic patients suffering baclofen associated neurotoxicity were treated by CAPD at our hospital. We obtain 12 cases with detailed clinical course of baclofen-associated neurotoxicity treated by hemodialysis from a literature review using the Medline and Science Citation Index, six of these patients were treated by early hemodialysis which was defined as hemodialysis intervention within 48 h of the onset of clinical toxicity. RESULTS: Our cases regain full consciousness within 2-3 days after the onset of neurotoxicity. Clinical characteristics of our cases including age, dialysis time, preexisting central nervous system (CNS) lesion, concomitant use of CNS depressant, total baclofen dose, onset of neurotoxicity, and duration of neurotoxicity are not significantly different from reported cases that treated by either early or routine hemodialysis. CONCLUSIONS: Baclofen should be avoided in uremic patients. When consciousness disturbance occurs in uremic patients, baclofen-associated neurotoxicity should be included in the list of differential diagnosis. According to experiences of our patients, we conclude that neither early nor routine hemodialysis is more effective than CAPD in shortening the recovery time of baclofen-associated neurotoxicity. PMID- 12739837 TI - Multiple renal cystic lesions and acute renal failure in non-Hodgkin's testicular lymphoma--a case report. PMID- 12739838 TI - P-ANCA positive renal vasculitis in association with renal cell carcinoma and prolonged hydralazine therapy. AB - Renal cell carcinoma and hydralazine drug therapy has each been reported as rare associations with pauci-immune renal vasculitis. We report a patient in whom both factors were operative simultaneously. A middle-aged man on long term hydralazine therapy presented with advanced renal failure. Serum was strongly positive for P ANCA with high anti-myeloperoxidase (MPO) titre and renal biopsy showed focal necrotizing glomerulonephritis. A renal mass was identified on scanning and at left nephrectomy a large renal cell carcinoma was removed. In spite of an apparently curative operation and discontinuation of hydralazine, P-ANCA remains strongly positive, he has had no recovery of renal function and has progressed to dialysis dependence. PMID- 12739839 TI - Donor calculi induced acute renal failure. AB - Nephrolithiasis is an infrequent complication following renal transplantation and acquisition of a stone with the donor kidney is rare. Indeed only a few cases of donor stones causing renal failure have been reported. Since the grafted kidney is denervated, stone disease may not present with the classic renal colic but rather with acute renal failure secondary to the obstruction. Identification of the precise cause is critical in order to avoid inappropriate therapy. We present our experience of two renal transplant patients who developed obstructive uropathy by stones originating from the donor kidneys. PMID- 12739840 TI - A Behcet case with renal amyloidosis. AB - Behcet's disease is a chronic systemic vasculitis with unknown etiology characterized by recurrent oral and genital ulcerations, ocular inflammation,having manifestations related to the skin and joints. Neurologic, pulmonary and gastrointestinal findings can also be observed during the course of the disease. In this report a case of Behcet's disease that had amyloidosis due to renal parenchymal involvement of the disease showing itself by nephrotic syndrome is presented. PMID- 12739841 TI - Spinal cord infarction following central-line insertion. AB - Hemothorax is a recognized complication of central line insertion into the jugular or subclavian vein. We describe a case of hemothorax consequent upon acute dialysis catheter insertion, which resulted in spinal cord infarction and quadriplegia. We postulate that the extensive mediastinal shift induced after insertion of the catheter resulted in stretching of the veins draining the cord with a resultant drop in perfusion pressure and infarction. This case highlights a hitherto unreported complication of this procedure. PMID- 12739842 TI - Early polysensorial enrichment: specific experience-induced structural changes in the parieto-occipital cortex of the rat. AB - The present research has been planned to determine the effects of early polysensorial enrichment on the plastic mechanisms that specify the complex organization of the cortical plate. The basal dendritic field, which is the input side of the neuron, developed increased geometrical complexity in enriched pups. Particularly striking was the specific territorial expansion of dendritic fields, induced by the exposition to a polysensorial enriched environment, observed in the lateral aspect of the parieto-occipital cortex. This implies an interplay involving the lateral aspect of the parieto-occipital cortex with afferents from whiskers and peribucal structures (areas for exploring the habitat). It is evident that during the early postnatal period the cerebral cortex develops a highly sophisticated network, which exhibits a high degree of substrate specificity tightly regulated by environmental cues. PMID- 12739843 TI - Expression of growth factors in fetal human olfactory mucosa during development. AB - The olfactory epithelium is one of the unique sites in the mammalian nervous system at which there is continual neurogenesis. Constant turnover of primary sensory neurons in the periphery results in remodeling of neuronal circuits and synapses in the olfactory bulb throughout life. Most of the specific mechanisms and factors that control this process are still unknown. Recent studies suggest that growth factors and their receptors may play a significant role in the development and continuous regeneration of olfactory neurons. In this study, standard immunohistochemical methods were used to detect the presence of epidermal growth factor-receptor, transforming growth factor-alpha and nerve growth factor-beta in the human olfactory epithelium of different developmental stages (7 to 30 weeks) in formalin-fixed tissue specimens. Immunoreactivity to epidermal growth factor-receptor was seen in basal cells, supporting cells, neurons, apical cytoplasmic region of olfactory epithelium, lamina propria and gland's cell cytoplasm. Similarly transforming growth factor-alpha immunoreactivity was recognized in basal cells, supporting cells, apical cytoplasmic region of olfactory epithelium, lamina propria and gland's cell cytoplasm but not in neurons. Nerve growth factor-beta detection was restricted on olfactory nerve cells, dentritic knob and basal cell layer. So, this study confirms the development of human olfactory mucosa and shows epidermal growth factor-receptor, transforming growth factor-alpha and nerve growth factor-beta proteins are reliable markers for developing olfactory epithelium. PMID- 12739844 TI - Inter-organ unevenness and catch-up growth in rats. AB - The recovery in body weight (catch-up growth) was studied in juvenile male and female rats either exposed to hypoxia during the first postnatal week (10% inspired O2, Hypoxic) or with limited caloric intake (by growing in large litter size) during the first two postnatal weeks (LowCal). Compared to controls, the experimental rats had a severe stunting in body growth. The effects on the dry and wet weights of the internal organs (kidney, lungs, brain, heart, liver, thymus, skeleton) differed between Hypoxic and LowCal, but were similar between sexes. Catch-up growth during the following weeks was more apparent in females than in males, and accompanied by increased oxygen consumption. At 1 and, especially, 3 months, the organs had almost recovered their original weights, although differences in protein and DNA content (measured in brain, lungs, heart, brown adipose tissue) were still present in some cases. In general, organ recovery was more apparent in LowCal than in Hypoxic, and in females than in males. In the experimental groups the unevenness in weight among organs (coefficient of variation of the individual weights relative to control) at 10 days, 1 month and 3 months correlated with the specific growth rate of the first, second and third month. This finding supports the possibility that inter-organ unevenness may represent an additional factor controlling the rate of catch-up growth. PMID- 12739845 TI - Dynamics of relative growth rate in Japanese quail lines divergently selected for growth and their control. AB - This study was undertaken to test the hypothesis that relative growth is the same in Japanese quail divergently selected for 4-week body weight for 30 generations, and their control. Relative growth rate is the increase in body weight per unit of body weight per unit of time, and it represents the efficiency of an animal as a producer of new body mass material. The maximum relative growth rate of the divergent and control lines were observed during the first week. Selection for high 4-week body weight has resulted in higher relative growth rate for the first two weeks compared to quail from the low line, but by week 3, quail from the low line had higher relative growth rate compared to quail from the high line. Asymptotic body weight increased by about 100 and 70%, respectively in the male and female high lines and declined by about 25% in the low lines. Absolute growth rate is thought to be related to the cumulative growth already achieved whereas relative growth rate is related to the amount of growth remaining. However, selection on absolute weight basis can elicit a response in relative growth rate in the early part of the developmental period which possibly pre-determines the asymptotic body weight. Since relative growth rate depends on the amount of growth remaining, the rate of decline in relative growth is then set accordingly. Furthermore, it is thought that different sets of genes may operate between the developmental period and maturation period. PMID- 12739846 TI - Future AAC technology needs: consumer perspectives. AB - This article suggests the need for rethinking the role that consumer perspectives will play (and ought to play) in the design and development of future augmentative and alternative communication (AAC) technologies. After carefully defining what they mean by "AAC consumers," "consumer perspectives," and "AAC technologies," the authors consider the extent to which current research has illuminated our understanding of (1) how consumers view AAC technologies and (2) how consumers themselves would define future technology needs. It's not a pretty picture. While it is clear that there already exist specific constructs and methods that could enable researchers and developers to learn more about consumer perspectives, it is equally clear that few, if any, researchers make much use of these constructs and methods. The authors focus specifically on ways that consumer perspective can helpfully infuse participatory action research, technology transfer processes, and ergonomics. In addition, the authors consider how research that investigates the ways in which assistive technologies impact the daily lives of individuals with severe communication impairment also influences public policy issues, which can in turn affect future definitions of AAC technology needs. Throughout the article, the authors advocate for a paradigm shift in the quantity and quality of the collaborations that occur between AAC consumers and AAC researchers, manufacturers, and developers. Augmented communicators and their families have a major stake in all types of research that can affect the design and development of AAC devices and accessories, and the results of this research, in turn, can affect public policy decisions about AAC technologies. This makes it all the more important that AAC consumers be involved at each step of the research and development process. PMID- 12739847 TI - Improving the design of augmentative and alternative technologies for young children. AB - If children with significant communication disabilities are to attain the power of language, literacy, and communication early in their development, they must have access to appropriate augmentative and alternative communication (AAC) systems. AAC technologies for young children should (a) be highly appealing, capturing, and sustaining of their interest; (b) be seamlessly integrated into all aspects of daily living; (c) provide access to the magical power of communication, language, and literacy; (d) be synchronous with children's diverse cultural and linguistic backgrounds; (e) be easy for children to learn and effortless for children to use; (f) grow easily and seamlessly with children as they develop; and (g) be efficient for families and professionals to learn, maintain, and develop. The design specifications to realize these requirements may vary across children given the diversity of skills, characteristics, and interests. This article summarizes what we know about young children and AAC technologies as related to each of these requirements and also discusses priorities to improve the design of AAC technologies in order to maximize outcomes for young children with significant communication disabilities. PMID- 12739848 TI - Improving AAC use for persons with acquired neurogenic disorders: understanding human and engineering factors. AB - The first section of this report focuses on those factors that influence augmentative and alternative communication (AAC) use by adults with acquired disability, including the natural course of the acquired neurogenic condition, use of multiple modes of communication, changing capabilities, communication patterns of adults, changing social roles, transitioning among environments, attitudes of AAC users and others, and instructional constraints. The second section provides recommendations about future technical and intervention innovations involving message management, alternative access strategies, and learning demands. PMID- 12739849 TI - AAC performance and usability issues: the effect of AAC technology on the communicative process. AB - In this article, Clark's theory of language use is applied to the study of conversations of augmented speakers and their addressees. Discussion focuses on how shared meaning--called common ground--is achieved, the process of grounding utterances under real-time constraints, and how the media characteristics of devices affect the grounding process. A joint action analysis of grounding will be applied to examples of word-board and Voice Output Communication Aid (VOCA) mediated conversations to show how participants organize their talk around specific media constraints to conduct their conversations and solve communication problems. The authors argue that this model of device-mediated communication performance, or some variety of it, has the potential to reconcile many of the individual research findings in this field within a single explanatory framework. If successful, this approach could be used to evaluate future research with an empirically-based model of communication performance. PMID- 12739850 TI - Enhancing participation in employment through AAC technologies. AB - Despite significant challenges, there are a growing number of documented cases of employment success for individuals who use augmentative and alternative communication (AAC). We review recent research on employment and AAC, and discuss the results within a framework that describes what is known about the worker, the workplace, technology, and society in achieving employment success for individuals who use AAC. Information on goals for future research and technology development is also provided. PMID- 12739851 TI - Enhancing literacy development through AAC technologies. AB - There is a critical need to understand teaching and technology supports that enable students who use augmentative and alternative communication (AAC) systems to engage in meaningful literacy experiences and foster conventional literacy skills. To thrive in classroom environments, they must have access to tools that can support them in active and independent literacy learning. These students need technology that allows them to move seamlessly between reading, writing, and communicating. They require technology that takes into account access needs, individual learning needs, the learning demands of technology, and literacy development across grades. Families and school teams need information that will assist them in providing the best tools and the most appropriate content within these tools throughout the school day. Teachers need information that supports them in providing exemplary literacy instruction to students who use AAC systems. This article explores and summarizes factors impacting literacy learning, including literacy capabilities of school-age students who use AAC, communication in literacy learning and use, reading and writing instruction in general education classrooms, and technology to support literacy learning. It is important that future technology tools provide a platform for levels of literacy learning: The power of technology will be reflected in its ability to provide access to and display the right content at the right time for students who use AAC. This article summarizes current factors thought to influence literacy learning and discusses priorities for future research and technology development. PMID- 12739852 TI - Connecting AAC devices to the world of information technology. AB - Modern day information technology (IT) is converging around wireless networks. It is now possible to check E-mail and view information from the World Wide Web from commercially available mobile phones. For individuals with disabilities, the ability to access multiple and different types of information not only promises convenience, but also can help to promote independence and facilitate access to public and private information systems. There are many barriers to access for people with disabilities, including technological hurdles, security, privacy, and access to these emerging wireless technologies. However, legislation, advocacy, standards, and research and development can ensure that users of augmentative and alternative communication (AAC) and assistive technology have access to these technologies. This article provides a historical context for the field of AAC and IT development, a review of the current state of these technologies, a glimpse of the potential of wireless information access for the lives of AAC users, and a description of some of the barriers and enablers to making access available to users of AAC and assistive technologies. PMID- 12739853 TI - Retinoids, LRAT and REH activities in eggs of Japanese quail following maternal and in ovo exposures to 3,3',4,4'-tetrachlorobiphenyl. AB - Environmental monitoring for the effects of organochlorine contaminants has identified changes in egg yolk retinoid concentrations, however underlying mechanisms are unknown. Breeding female Japanese quail, Coturnix coturnix japonica, received 3 bimonthly injections of a low dose (5 microg/g) of the coplanar polychlorinated biphenyl (PCB) 3,3',4,4'-tetrachlorobiphenyl. No overt maternal toxicity or embryotoxicity was detected. For the exposed females, oocytes (yolk deposition nearly completed) and freshly laid eggs contained higher concentrations of retinol. For PCB-exposed female quail, freshly laid eggs and fertile eggs incubated for 6 days contained higher concentrations of retinyl palmitate. These results suggest that the PCB congener caused greater deposition of retinoids in yolks during late oogenesis or greater net esterification in the yolk-sac membrane. Retinyl ester hydrolase (REH) activity, assayed in the microsomal fraction of the yolk-sac membrane, was significantly inhibited in the PCB-exposed group. Therefore, the higher retinyl palmitate concentrations may be explained by less retinyl palmitate conversion to retinol. In a second experiment, fertile quail eggs were injected with 2, 10 or 20 microg of the PCB congener and incubated for 6 days. In contrast to the maternal exposure, yolk retinol decreased and retinyl ester hydrolase increased in the PCB-injected eggs. The difference in the results of adult versus egg injections may be related to several factors including adult transformation of the PCB to toxic metabolites and differences in post-transcriptional regulation of REH expression. PMID- 12739854 TI - Lead poisoning in upland-foraging birds of prey in Canada. AB - We examined the degree of lead exposure, based on tissue-lead concentrations, in 184 raptors of 16 species found dead across Canada. The most prevalent species available for examination were Red-tailed hawks, Great horned owls, and Golden eagles (n = 131). The majority of individuals examined had very low lead accumulation, however 3-4% of total mortality in these 3 most commonly encountered species was attributed to lead poisoning. In addition, 1 of 9 Bald Eagles found dead far from aquatic environments was lead poisoned; and a single Turkey Vulture had a highly elevated bone-lead concentration (58 microg/g dry weight). Evidence from our study, along with other published research, indicates that upland-foraging birds of prey and scavengers that typically include game birds and mammals in their diets, are at risk for lead poisoning from the ingestion of lead projectiles from ammunition used in upland hunting. The use of non-lead ammunition for hunting upland game would effectively remove the only serious source of high lead exposure and lead poisoning for upland-foraging raptors. PMID- 12739855 TI - Using chorioallantoic membranes for non-lethal assessment of persistent organic pollutant exposure and effect in oviparous wildlife. AB - David Peakall and co-workers pioneered innovative approaches that utilized extra embryonic membranes to assess accumulation of organochlorine pesticides in eggs. This technique provided the foundation for an entire line of research to improve non-lethal methods for assessing contaminant exposure in oviparous wildlife. Currently, analysis of chorioallantoic membranes (CAMs) provides predictable estimates of chlorinated contaminant presence in eggs and in maternal tissues. Field studies have been conducted with herons, stilts, alligators, crocodiles, and sea turtles. Controlled dose-response studies have been completed in chickens. The following manuscript presents the foundations for the CAM approach and a review of research findings involving this technique. PMID- 12739856 TI - Trace elements in lesser scaup (Aythya affinis) from the Mississippi flyway. AB - Previous research reported that concentrations of selenium in the livers of 88 95% of lesser scaup from locations in Lake Erie, Lake St. Clair, and Lake Michigan, USA were either elevated (10-33 microg/g dry weight [dw]) or in the potentially harmful range (> 33 microg/g dw). In order to determine the geographic extent of these high selenium concentrations, we collected lesser scaup in Louisiana, Arkansas, Illinois, Minnesota, Wisconsin, and Manitoba and analyzed the livers for 19 trace elements. We found that all trace element concentrations, except for selenium, generally were low. Arsenic, which usually is not detected in liver samples, was detected in Louisiana and may be related to past agricultural usages. Chromium, which also is not usually detected, was only present in lesser scaup from Arkansas and may be related to fertilizer applications. Cadmium and mercury concentrations did not differ among locations and concentrations were low. Selenium concentrations in Arkansas (geometric mean = 4.2 microg/g dw) were significantly lower than those in Louisiana (10.7 microg/g dw), Illinois (10.5 microg/g dw), and Minnesota (8.0 microg/g dw); concentrations in Wisconsin and Manitoba were intermediate (6.6 and 6.5 microg/g dw). About 25% of lesser scaup livers contained elevated selenium concentrations; however, none were in the harmful range. We concluded that selenium concentrations in lesser scaup in the Mississippi Flyway are elevated in some individuals, but not to the extent that has been documented in the industrial portions of the Great Lakes. PMID- 12739857 TI - Development and validation of a herring gull embryo toxicokinetic model for PCBs. AB - A toxicokinetic model was developed to describe polychlorinated biphenyl (PCB) accumulation by herring gull (Larus argentatus) embryos during development. The model consists of two components, a bioenergetics model that predicts the lipid mass balance of embryo and yolk compartments with time and an empirical toxicokinetic model that describes PCB partitioning between lipid compartments in the egg. The model was calibrated using data on PCB and lipid partitioning between embryo and yolk + albumen at four time points during incubation in herring gull eggs injected with a PCB mixture, combined with data sets on herring gull embryo growth rates and bioenergetic demands with time. The model was validated using independent data consisting of maternally exposed, field incubated Lake Superior herring gull eggs that varied in incubation ages over the range of 8.5 d to pipping age (26-28 days). PCB concentrations in 6-9 d embryos were nearly an order of magnitude less than predicted by equilibrium lipid partitioning between the embryo and yolk + albumen compartments of the eggs. PCB concentrations in embryos were adequately predicted by equilibrium partitioning, however, for eggs incubated for 23-28 d. An empirical relationship was developed to account for the apparent nonequilibrium behaviour of PCBs during early development. The model was sensitive to the mass of yolk lipids and the mass of PCBs deposited to fresh eggs and much of the variability in embryo PCB concentrations could by explained by accounting for variability in these input parameters. Consistent with experimental data for other avian species, the model predicts that the highest PCB concentrations in the embryo/chick occur during pipping or soon after when yolk lipids have been completely resorbed by the embryo. PMID- 12739858 TI - Common loon eggs as indicators of methylmercury availability in North America. AB - Increased anthropogenic mercury (Hg) deposition since pre-industrial times, and subsequent transformation of inorganic Hg to methylmercury (MeHg) in aquatic environments, has created areas in North America where Hg poses a relatively high risk to wildlife, especially long-lived, piscivorous species. From 1995 to 2001, we opportunistically collected 577 eggs abandoned by Common Loons from eight states. Egg-Hg concentrations ranged from 0.07 to 4.42 microg/g (ww) or 0.10 to 19.40 microg/g (dw). Mercury was higher in eastern than in western North America. Female blood-Hg concentrations strongly correlated with those of eggs from the same territory even though the mean intraclutch Hg difference was 25%. In New England, egg volume declined significantly as egg-Hg concentrations increased. Fertility was not related to egg-Hg concentrations. Based on existing literature and this study's findings, egg-Hg risk levels were established and applied to our US data set and an existing Canadian data set. Regionally, we found the greatest risk levels in northeastern North America. With few exceptions, loon eggs are suitable indicators of methylmercury availability on lakes with territorial pairs. PMID- 12739859 TI - Bioaccumulation patterns and temporal trends of mercury exposure in Wisconsin common loons. AB - A long term field study was initiated by the Wisconsin Department of Natural Resources in 1992 to elucidate patterns of common loon (Gavia immer) mercury (Hg) exposure. Analysis of loon blood and feather samples collected from recaptured adult loons in Wisconsin 1992-2000 found evidence of a decline in overall body burdens of mercury in common loons for this region. The interval between sampling individual loons spanned 2-8 years, a sufficient length of time to observe a change in tissue Hg concentrations. Loon chick blood Hg levels declined by 4.9% annually for chicks sampled on 33 lakes during the period 1992-2000. This is the first evidence we are aware of showing a recent regional annual decrease in common loon Hg exposure. Repeated captures of wild loon chicks in Wisconsin shows that blood Hg concentrations can increase during the period of rapid feather growth (weeks 2-5), although the rate of increase is very slow. Mean egg Hg levels ranged from 0.19 to 0.87 microg Hg/g wet weight (ww) in samples collected 1996-2000. Egg Hg concentration was inversely and significantly related to lake pH (p < 0.0001; r2 = 0.55). Adults and chicks were often captured simultaneous during the period 1992-2000. Correlations were highest between sibling blood Hg levels (r = 0.88) and chick blood and adult blood (male r = 0.61, female r = 0.52) Hg levels, likely reflecting the influence of the Hg content of prey from the natal lake on loon blood Hg levels. The relationship between feathers and blood of adults and that of chick blood and adult feathers was weaker. PMID- 12739860 TI - Influence of food supply and chlorinated hydrocarbon contaminants on breeding success of bald eagles. AB - Food supply and contaminants were investigated as possible causes of low bald eagle productivity near a bleached kraft pulp and paper mill at Crofton on Vancouver Island, British Columbia. Over a seven year period, 1992-1998, average productivity of five eagle territories situated south of the pulp mill at Crofton was significantly lower (0.43 young/occupied territory) than six territories north of the mill (1.04 young/occupied territory). A reference population of 32 territories located in Barkley Sound on the west coast of Vancouver Island demonstrated intermediary mean productivity (0.75 young/occupied territory). Measures of prey biomass delivered to nests were lowest south of the mill, and correlated significantly with nesting success. On average, measures of energy delivered to nests and a parameter determined to be related to prey availability, adult nest attendance time, accounted for about 70% of variability in nest success. Contaminant concentrations, including pulp mill derived polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs), as well as dichlorodiphenyldichloroethane (DDE), polychlorinated biphenyls (PCBs), and calculated tetrachlorodibenzo-p-dioxin toxic equivalents (TEQs) were significantly greater in plasma samples of nestlings from south of the mill compared to the other two sites, but did not correlate significantly with individual nest success data. Nests south of the mill concentrate around Maple Bay, which appears to be a deposition area for contaminants transported by tides and currents from sources such as the pulp mill. Concentrations of DDE and PCBs in plasma of nestling eagles from south of the mill were less than the critical values estimated to affect production of young. For TEQs, there are no published critical values for plasma by which to compare our results. We conclude that less than adequate energy provisioning to nests, presumably related to low prey availability, was likely the main cause of poor nest success south of the mill site at Crofton. However, higher concentrations of both DDE and PCDD/F derived TEQs may have acted in concert with food stress to further reduce bald eagle productivity. PMID- 12739861 TI - An assessment of DDT and other chlorinated compounds and the reproductive success of American robins (Turdus migratonrius) breeding in fruit orchards. AB - Although DDT was banned in the 1970s, American robins (Turdus migratorius) breeding in fruit orchards of the Okanagan Valley, British Columbia, continue to be contaminated with DDT and its metabolites. The objectives of our study were (1) to assess organochlorine (OC) contamination in robins breeding in Okanagan orchards (1993-1995, 1997-1998) and (2) to determine if exposure affected reproductive success when compared to robins from non-orchard habitat (lower mainland, British Columbia). Robins in orchards had total DDT egg residues of 48.64 mg/kg (geometric mean; n = 92) while those in non-orchard habitat had 1.10 mg/kg (geometric mean; n = 26), wet weight. The probability of nest survival during the incubation period was 96.7% (confidence interval: 95.7-97.5%; n = 165) in orchard habitat and 96.7% (confidence interval: 94.6-98.1%; n = 28) in non orchard habitat. During the nestling period the probability of nest survival was 98.2% (confidence interval: 97.2-98.9%; n = 123) in orchard habitat and 96.2% (confidence interval: 92.8-98.0%; n = 34) in non-orchard habitat. Clutch (p < 0.0001) and brood size (p = 0.0133) were larger in orchards (n = 150 and n = 93, respectively) compared to non-orchard nests (n = 42 and n = 23, respectively) with no difference in fledge rate. DDE (r2 = 0.11, p = 0.0030, n = 68) and dieldrin (r2 = 0.29, p < 0.0001, n = 68) were negatively correlated with fledge rate in robin eggs collected from orchard habitat, however, low r2 values signify minimal biological significance. Although American robins nesting in Okanagan orchards are exposed to high OC levels, reproductive success does not appear to be negatively impacted. PMID- 12739863 TI - Assessing temporal trends in contaminants from long-term avian monitoring programs: the influence of sampling frequency. AB - The effect of sampling frequency on the detection of statistically significant temporal trends in egg contaminant levels was examined using data from the Great Lakes Herring Gull Monitoring Program. Decreased identification of statistically significant trends was apparent in the sampling regimes where samples were collected less frequently. When statistically significant declines were observed, sampling at two and four year intervals resulted in the trend being identified later than with annual monitoring. The design of monitoring programs must balance costs and data quality. Programs should ensure that the data collected are adequate to address critical questions. PMID- 12739862 TI - Gonadal feminization and halogenated environmental contaminants in common terns (Sterna hirundo): evidence that ovotestes in male embryos do not persist to the prefledgling stage. AB - Common terns (Sterna hirundo) and roseate terns (Sterna dougallii) breed on Bird Island, Massachusetts, USA, near a Superfund site highly contaminated with polychlorinated biphenyls (PCBs). Observations of skewed sex ratios and female female pairings among endangered roseate terns (Nisbet and Hatch (1999) Ibis 141, 307) suggested the possibility of contaminant-related endocrine disruption in these birds and prompted investigation of common terns as a surrogate species. In 1993 and 1994, 60-90% of pipping male common tern embryos sampled exhibited ovarian cortical tissue in their testes (ovotestes) (Nisbet et al. (1996) Bull. Environ. Contam. Toxicol. 57, 895; Hart et al. (1998) Mar. Environ. Res. 46, 174). To examine the possible impact of ovotestes on the reproductive capabilities of common terns, we examined gonadal histology in common tern prefledglings (approximately 21 days old) collected from Bird Island in 1995. As a measure of embryonic contaminant exposure, contaminants were measured in a subset of eggs collected from the same nests as the prefledglings. Concentrations of total PCBs in these eggs ranged from 14.4 to 546 microg/g lipid. No evidence of ovotesticular development was observed in any of the 19 male prefledglings examined. Some gonadal irregularities were observed, including small nodules of testicular tissue within the epithelial capsule of the testes, but these were judged not likely to affect testicular function. There was no relationship between any observed irregularities and levels of contaminants present in the matched eggs. The results suggest that the ovotestes that occur in 60-90% of pipping common tern embryos from this site become fully regressed by approximately 21 days posthatch. Our data from this and previous studies are consistent with the idea that ovotestes occur naturally in some individual common terns at hatching, although the frequency of occurrence may be enhanced by exposure to chlorinated organic contaminants such as PCBs. In either case, we suggest that the presence of ovotestes in common tern embryos from PCB contaminated sites such as Bird Island does not lead to permanent alterations in gonadal histology that would be expected to impair reproductive function. PMID- 12739864 TI - Hepatic EROD activity is not a useful biomarker of polychlorinated biphenyl exposure in the adult herring gull (Larus argentatus). AB - Liver concentrations of polychlorinated biphenyls (PCBs) and chicken embryo hepatocyte (CEH) bioassay-derived 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TCDD-EQs) were measured in livers of adult herring gulls (Larus argentatus) collected from several locations on the Great Lakes and two reference sites. Total PCB concentrations (sum of 42 congeners) and TCDD-EQ concentrations were compared with hepatic ethoxyresorufin-O-deethylase (EROD) activity, methoxyresorufin-O-deethylase (MROD) activity and immunodetectable cytochrome P4501A (CYP1A) protein concentration. EROD and MROD activity were not significantly correlated with total PCB concentration or TCDD-EQ concentration in liver tissue. CYP1A protein concentration was significantly correlated with total PCB concentration, but the linear relationship had little predictive power. We conclude that EROD is not a useful biomarker of PCB exposure in the adult herring gull. PMID- 12739865 TI - Sensitivity of bald eagle (Haliaeetus leucocephalus) hepatocyte cultures to induction of cytochrome P4501A by 2,3,7,8-tetrachlorodibenzo-p-dioxin. AB - Graded doses of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) were added to primary hepatocyte cultures of bald eagle (Haliaeetus leucocephalus) embryos to determine their sensitivity to induction of cytochrome P4501A (CYP1A) and porphyrin accumulation. No porphyrin accumulation was observed, but both CYP1A catalytic activity (using the ethoxyresorufin-O-deethylase (EROD) assay) and immunodetectable CYP1A were induced by relatively high concentrations of TCDD. Bald eagle hepatocytes were less sensitive to CYP1A induction than hepatocytes from any other avian species that we have studied to date. These in vitro results are in general agreement with recent assessments of field data, which indicate that bald eagles are relatively insensitive to some of the effects of TCDD and related compounds. Preparation of bald eagle hepatocytes was challenging because existing methods did not yield monolayers of cells. Here we describe details of a new method that was successful for bald eagle hepatocytes. This new method is used routinely in our laboratory to prepare hepatocyte cultures from birds for examination of various biochemical responses to environmental contaminants. PMID- 12739866 TI - Effects of methyl mercury exposure on the growth of juvenile common loons. AB - We conducted a dose-response laboratory study to quantify the level of mercury exposure associated with negative effects on the development of common loon chicks reared in captivity from hatch to 105 days. A dose regimen was implemented that provided exposure levels that bracketed relevant exposure levels of methyl mercury found in loon chicks across North America. We observed no overt signs of mercury toxicosis and detected no significant effect of dietary mercury exposure on growth or food consumption. However, asymptotic mass was lower in chicks that hatched from eggs collected from nests on low pH lakes relative to eggs from neutral pH lakes. Rapid excretion of methyl mercury during feather growth likely provides loon chicks protection from methyl mercury toxicity and may explain the lack of convincing toxicological findings in this study. Lake-source effects suggest that in ovo exposure to methyl mercury or other factors related to lake pH have consequences on chick development. PMID- 12739867 TI - Biological effects of marine PCB contamination on black guillemot nestlings at Saglek, Labrador: liver biomarkers. AB - Black guillemots (Cepphus grylle) in Saglek Bay, Labrador have elevated polychlorinated biphenyl (PCB) concentrations due to marine sediment contamination around a former military site. We measured liver biomarkers and sigmaPCB concentrations in 31 nestlings from three PCB-exposure groups: Reference group (range: 15-46 ng/g liver, wet wt.), moderately exposed Islands group (24 150 ng/g), and highly exposed Beach group (170-6200 ng/g). Biomarker responses were dose-dependent and in some cases sex-dependent. Livers of female Beach nestlings were enlarged 36% relative to Reference females. In both sexes, Beach nestlings had liver ethoxyresorufin-O-deethylase (EROD) activities elevated 79% and liver retinol concentrations reduced 47%. Retinyl palmitate concentrations were reduced 50% but only among female nestlings. Island nestlings also exhibited EROD induction (57%) and reductions in retinol and retinyl palmitate concentrations (28 and 58%, respectively). Liver lipid content increased with sigmaPCBs in both sexes, and correlated with liver mass in males. Malic enzyme activity and porphyrin concentrations showed little association with sigmaPCBs. Although similar associations between liver biomarkers and organochlorine exposure in fish-eating birds are well documented, typically exposures involve multiple contaminants and there is uncertainty about specific PCB effects. Our findings indicate that liver biomarkers respond to relatively low PCB exposures (approximately 73 ng/g liver) in guillemots. PMID- 12739869 TI - Chlorinated hydrocarbon contamination in osprey eggs and nestlings from the Canadian Great Lakes basin, 1991-1995. AB - Populations of osprey (Pandion haliaetus) in the Great Lakes basin declined dramatically during the 1950s-1970s due largely to adverse effects of persistent chlorinated hydrocarbons, ingested in their fish prey, on eggshell thickness and adult survival. Nevertheless, these contaminants were not measured in osprey tissues during the decades of decline on the Canadian Great Lakes. Between 1991 and 1995, we monitored recovering osprey populations on the Great Lakes, including Georgian Bay and the St. Marys River area on Lake Huron and the St. Lawrence Islands National Park, as well as at two inland sites within the basin. Current OC levels, even from the most contaminated lakes, were typically lower than those associated with reproductive effects. DDE levels in fresh eggs averaged 1.2-2.9 microg/g, well below the 4.2 microg/g level associated with significant eggshell thinning and shell breakage. Nevertheless, a proportion of eggs from all study areas did exceed this level. PCB levels in eggs seldom exceeded 5 microg/g except in one lake of high breeding density in the Kawartha Lakes inland study area, where the mean sum PCB level was 7.1 microg/g and the maximum concentration measured was 26.5 microg/g. On average, mean reproductive output (0.78-2.75 young per occupied nest) of breeding populations in Great Lakes basin study areas exceeded the threshold of 0.8 young thought necessary to maintain stable populations. We concluded that, although eggs and especially nestling plasma, are useful in reflecting local contaminant levels, ospreys are relatively insensitive, at least at the population level, to health effects of current levels of chlorinated hydrocarbons on the Canadian Great Lakes. PMID- 12739868 TI - Effects of dietary PCB exposure on adrenocortical function in captive American kestrels (Falco sparverius). AB - We experimentally examined the effects of dietary exposure to polychlorinated biphenyls (PCBs) on adrenocortical function in American kestrels (Falco sparverius). Nine captive male American kestrels previously exposed to a PCB mixture (Aroclor 1248:1254:1260; 1:1:1) in their diet were subjected to a standardized capture, handling and restraint protocol designed to produce an increase in circulating corticosterone. A similar protocol has been applied to a wide range of avian species and was used here to evaluate the response of PCB exposed and control kestrels to a defined physical stressor. Both baseline and stress-induced corticosterone levels were significantly lower in PCB-exposed birds when compared with control birds of the same age. PCB-exposed birds exhibited significantly lower corticosterone levels during the corticosterone response when compared with control birds, independent of body condition. Furthermore, baseline corticosterone concentrations exhibited a hormetic response characterized by an inverted U-shaped dose response in relation to total PCB liver burden. These results support several recent studies which report decreased levels of circulating corticosterone in PCB-exposed wild birds. The results presented here provide the first evidence that exposure to an environmentally relevant level of PCBs (approximately 10 mg/kg body weight) can impair the corticosterone stress response in kestrels, potentially increasing the susceptibility of birds to environmental stressors such as severe weather and predatory and human disturbance. PMID- 12739870 TI - Biomarker responses in nesting, common eiders in the Canadian arctic in relation to tissue cadmium, mercury and selenium concentrations. AB - Populations of many North American sea ducks are declining. Biomarkers may offer valuable insights regarding the health and fitness of sea ducks in relation to contaminant burdens. In this study we examined body condition, immune function, corticosterone stress response, liver glycogen levels and vitamin A status in relation to tissue concentrations of mercury, selenium and cadmium in female common eiders during the nesting period. The study was conducted in the eastern Canadian arctic during July, 2000. Hepatic mercury, selenium and renal cadmium concentrations ranged 1.5-9.8, 6.5-47.5 and 74-389 microg/g, dry wt, respectively. Mercury concentrations were negatively related to dissection body mass, heart mass and fat mass. Cadmium concentrations were negatively related to mass at capture and dissection mass after controlling for the mercury concentration-dissection mass relationship. Cell-mediated immunity was assessed by the skin swelling reaction to an injection of phytohemagglutinin-P, and was unrelated to metal concentrations. After adjusting the corticosterone concentration to account for the time between capture and sampling, there was a negative relationship between the residual corticosterone concentration and selenium. Liver glycogen concentrations were not significantly related to metal concentrations. Mercury concentrations were positively related to those of hepatic retinol and retinyl palmitate and the ratio of the retinol to retinyl palmitate in liver. They were negatively related to the ratio of plasma to liver retinol. Our findings do not indicate that exposure to metals may have adversely affected the health of these birds. They do, however, suggest that more research is required to elucidate mechanisms by which exposure to these metals could impact body condition. PMID- 12739871 TI - In vitro biomonitoring in polar extracts of solid phase matrices reveals the presence of unknown compounds with estrogenic activity. AB - Determination of estrogenic activity has so far mainly concentrated on the assessment of compounds in surface water and effluent. This study is one of the first to biomonitor (xeno-)estrogens in sediment, suspended particulate matter and aquatic organisms. The relatively polar acetone extracts from these solid phase matrices do not contain the well-known estrogenic compounds such as hormones, alkylphenols and phthalates. An in vitro 'estrogen receptor-mediated chemical activated luciferase gene expression' (ER-CALUX) assay was applied to samples from various locations in the Netherlands. Estrogenic activity measured in polar fractions of particulate matter and sediment extracts ranged from below detection limit to up to 4.5 pmol estradiol equivalents (EEQ)/g dry weight. Estrogenic activity in freshwater river sediments was up to five times higher compared to sediments from large lakes and coastal locations. Tissue extracts EEQs were determined in bream (Abramis brama), flounder (Platichthysflesus), freshwater mussels (Dreissena polymorpha) and marine mussels (Mytilus edulis). The highest biota EEQ levels were found in the freshwater zebra mussel (30 pmol EEQ/g lipid). One sample site showed greatly elevated EEQs in sediment and biota, which correlated with effects found in the wild populations of bream. The EEQ activity of the unknown compounds in the polar fraction mostly was much higher than the calculated EEQ levels based on known estrogens in the non-polar fraction (previously published data). PMID- 12739872 TI - Age-related differences in the sensitivity of the fish immune response to a coplanar PCB. AB - Polychlorinated biphenyls (PCBs) are widespread environmental pollutants. Because of their persistence and bioaccumulation in aquatic organisms (among other factors), the biological impact of PCB exposure on resident fish populations is of particular concern. To assess the effect(s) of an environmentally relevant coplanar PCB congener on the fish immune response, juvenile and aged Japanese medaka (Oryzias latipes) were injected i.p. with either vehicle or PCB 126 (at 0.01 or 1.0 microg/g BW) and examined after 3 and 14 days. CYP1A protein levels, examined as an indicator of PCB exposure, were significantly increased (compared to controls) in all fish treated with the highest PCB dose. Kidney phagocyte superoxide (O2*-) production was examined to indicate effects upon innate immune function. After 14 days, unstimulated O2*- production by kidney phagocytes from juvenile and aged medaka treated with the highest PCB dose was significantly increased compared to controls. Stimulated O2*- production by aged PCB-treated fish was unaffected (compared to controls) at both post-exposure timepoints. However, phagocytes from PCB-treated juvenile medaka demonstrated reduced O2*- production at 3 days post-exposure and increased levels after 14 days (compared to controls). These results demonstrate the sensitivity of medaka phagocyte function for examining PCB-induced immunotoxicity. PMID- 12739873 TI - Relative contributions of organochlorine contaminants, parasitism, and predation to reproductive success of eastern spiny softshell turtles (Apalone spiniferus spiniferus) from southern Ontario, Canada. AB - We examined hatching success, predation rates, rates of parasitism, sex ratio, and egg viability in eggs of the eastern spiny softshell (Apalone spiniferus spiniferus), a threatened species in Canada. Eggs were monitored from three populations, located at Thames River, Rondeau Provincial Park, and the Long Point National Wildlife Area, in southern Ontario, Canada in 1998. Concurrently, we measured organochlorine pesticides, PCBs, dibenzo-p-dioxins and furans from eggs from the same nests. Contaminant concentrations in eggs were similar among sites. There was no correlation between hatching success, parasitism and depredation rates, or the proportion of hatchlings that were males with total PCBs or individual pesticides, but there was a positive correlation between egg viability with concentrations in eggs of total PCBs, and with five pesticides. We found no evidence that the reproductive success of softshell turtles was compromised due to organochlorine contamination. The most important factors determining hatching success of eggs was predation, followed by egg viability and parasitism. PMID- 12739874 TI - Subtle effects of herbicide use in the context of genetically modified crops: a case study with glyphosate (Roundup). AB - Research on the effects of glyphosate and seed germination spans 30 years. Despite several studies reporting detrimental effects of the herbicide on seedling germination and growth, glyphosate is still being registered for use as a weed killer and preharvest desiccant. Its nonselective nature and low chance of species developing resistance has lead to the development of genetically modified crops tolerant to the herbicide which also raises concerns about increased reliance on herbicide use, and subtle ecological impact. This paper presents the result of a literature review on past studies mostly, on crop species, and the results of a new experiment performed with emphasis on noncrop species. The new experimental part attempted to determine whether glyphosate (Roundup) would have an effect on the germination and growth of the F1 generation of seeds produced by plants sprayed with the herbicide. It was designed to mirror spray drift which might affect noncrop plants in nontarget drift zones. Of the 11 species tested using treatments of 0% (control), 1%, 10% or 100% of a 890 g a.i./ha label rate solution sprayed near seed maturity, seven showed a significant effect of the glyphosate treatment on germination and/or growth characteristics. Results of this experiment together with several previous studies reviewed in this paper suggest that there are significant effects to keep in mind when using herbicides such as glyphosate as severe ecological changes could occur. PMID- 12739875 TI - Effects of endocrine modulators on sex differentiation in birds. AB - This mini-review focuses on sexual differentiation of the reproductive organs and the brain in birds and the effects of endocrine modulators on these processes. Sex determination in birds is genetically controlled, but the genetic events implicated are largely unknown. Female birds have one Z and one W sex chromosome, while males have two Z sex chromosomes. It is not clear whether it is the presence of the W chromosome in females, the double dose of the Z chromosome in males vis-a-vis females, or both of these characteristics that are crucial for the determination of sex in birds. Oestradiol directs sexual differentiation in birds during critical periods of development. Consequently, exogenous compounds that interfere with the endogenous oestrogen balance can disrupt sexual differentiation of the reproductive organs and the brain. Therefore, sexual differentiation in birds provides a good model for studying the effects of endocrine modulators at various biological levels from gene expression to behaviour. Some compounds known to be present in the environment can alter endocrine function and have adverse effects when administered during development, resulting in alterations in gonads, accessory sexual organs, and behaviour. Data reviewed in this paper are mostly from laboratory studies on endocrine modulators with oestrogenic activity, whereas evidence for adverse effects of pollutants on sexual differentiation in avian wildlife is scarce. PMID- 12739876 TI - Review: porphyrins as biomarkers for hazard assessment of bird populations: destructive and non-destructive use. AB - In this review the biochemical, metabolic and toxicological significance of porphyrins in birds is examined, and their use as biomarkers of exposure to xenobiotics and heavy metals is explored. Laboratory studies pinpointing the main classes of compounds that alter porphyrin profiles are described, as well as those which defined the resulting porphyrin profiles and target organs. Field studies in which the biomarker was validated on natural populations of several species of birds are then reviewed. We finally illustrate their potential as a nondestructive biomarker suitable for situations in which destructive sampling of natural populations must be avoided, suggesting the implementation of this approach. PMID- 12739877 TI - Neurotoxic pesticides and behavioural effects upon birds. AB - Organochlorine, organophosphorus, carbamate, pyrethroid and neonicotinoid insecticides and organomercury fungicides are all neurotoxic and therefore have the potential to cause behavioural disturbances in birds. A number of studies have described behavioural effects caused to captive birds by neurotoxic pesticides, but it is very difficult to measure such effects in the field, which is a serous limitation given their potential to cause adverse effects at the population level. The mode of action, and the neurotoxic and behavioural effects of these compounds are briefly reviewed before considering evidence for their effects in the laboratory and field. Behavioural effects may cause adverse changes at the population level either directly or indirectly. Direct effects upon avian populations may be due to disturbances of reproduction, feeding, or avoidance of predation. Indirect effects on predators may be the consequence of direct action upon the prey population leading to either (1) reduction of numbers of the prey population, or (2) selective predation by the predator upon the most contaminated individuals within the prey population. Attention is given to the historic evidence for neurotoxic and behavioural effects of persistent organochlorine insecticides, raising the question of retrospective analysis of existing data for this once important and intensively studied class of compounds. Less persistent pesticides currently in use may also have neurotoxic effects upon birds in the field. Sometimes, as with some OPs, their effects may outlast the persistence of their residues, and the ecotoxicity and persistence of some may be affected by interactions with other environmental chemicals. The development of new mechanistic biomarker assays could improve understanding of behavioural effects and possible associated effects at the population level caused by such compounds in the field. PMID- 12739878 TI - Behavioural effects of pesticides in bees--their potential for use in risk assessment. AB - This paper reviews a wide variety of behavioural effects that have been reported in bees following exposure to pesticides, primarily insecticides. These range from effects on odour discrimination in the individual to the loss of foraging bees due to disruption of their homing behaviour. Some of these effects have the potential to have a significant impact on the development and survival of colonies. However, there is currently little guidance available on the types of behavioural data which should be collected during laboratory, semi-field or field regulatory studies or how they should be included and interpreted in risk assessment. Further work is required to allow risk assessment to include significant behavioural effects and their longer term consequences on colony survival and development. Such an approach will require a larger base set of data to predict the longer-term consequences on colonies of short-term effects on individuals, e.g. through population modelling. PMID- 12739879 TI - A proposal for the use of biomarkers for the assessment of chronic pollution and in regulatory toxicology. AB - Despite a wealth of information on biomarkers, they are not routinely used for regulatory purposes, even though the potential benefits of biomarkers to rationalise complex exposure-response relationships are clear. Biomarkers can be inappropriately applied or misinterpreted, because the fundamental assumptions in exposure-response relations have not been considered. Factors causing temporal and spatial variability in biomarker responses are reviewed. These include numerous geochemical and biotic variables. The variation can be minimised by appropriate study site selection, experimental replication, multivariate epidemiological approaches, normalised controls, and temporal calibration of responses; so that the regulatory use of biomarkers for biomonitoring and tracking pollution events, including chronic or multiple exposures to complex mixtures is possible. We propose and define the characteristics of biomarkers of chronic exposure or effect, which must measure changes in pollution/effect against long-term changes in other general stresses (disease, nutrition, environmental quality), relate to cumulative injury, and remain responsive over months or years. Neuroendocrine, immunological, and histological biomarkers are suggested for chronic pollution. We propose a regulatory framework for biomarkers based on a weight of evidence approach that can integrate biomarkers in risk assessment and long-term monitoring programmes. PMID- 12739881 TI - Use of retrospective data to assess ecotoxicological monitoring needs for terrestrial vertebrates residing in Atlantic coast estuaries. AB - The "Contaminant Exposure and Effects-Terrestrial Vertebrates" (CEE-TV) database contains 4,336 records of ecotoxicological information for free-ranging amphibians, reptiles, birds, and mammals residing in Atlantic and Florida Gulf coast estuaries and their drainages. To identify spatial data gaps, those CEE-TV records for which the specific study location were known (n = 2,740) were combined with watershed and wildlife management unit boundaries using Geographic Information Systems software. The US Environmental Protection Agency's Index of Watershed Indicators (IWI), which classifies watersheds based on water quality and their vulnerability to pollution, was used to prioritize these data gaps. Of 136 watersheds in the study area, 15 that are classified by the IWI as having water quality problems or high vulnerability to pollution lacked terrestrial vertebrate ecotoxicological monitoring or research in the past decade. Older studies within some of these watersheds documented high levels of contaminants in wildlife tissues. Of 90 National Wildlife Refuge units, 42 without current data fall within watersheds of concern. Of 40 National Park units larger than 1 km2, 17 without current data fall within watersheds of concern. Issues encountered in this analysis highlighted the need for spatially and temporally replicated field monitoring programs that utilize random sampling. Without data from such studies, it will be difficult to perform unbiased assessments of regional trends in contaminant exposure and effects in terrestrial vertebrates. PMID- 12739880 TI - Immunotoxicity of organophosphorous pesticides. AB - This study reviews the toxic effects of organophosphate (OP) pesticides on the immune systems and immune functions of invertebrates, fish, and higher vertebrate wildlife. The fundamental features and mechanisms of OP-induced immunotoxicity are illustrated with reference to parathion, chlorpyrifos, malathion, and diazinon. Immunotoxicity may be direct via inhibition of serine hydrolases or esterases in components of the immune system, through oxidative damage to immune organs, or by modulation of signal transduction pathways controlling immune functions. Indirect effects include modulation by the nervous system, or chronic effects of altered metabolism/nutrition on immune organs. Immunotoxicities are varied and include pathology of immune organs, and decreased humoral and/or cell mediated immunity. Altered non-specific immunity, decreased host resistance, hypersensitivity and autoimmunity are also features of immunotoxicity; although not all of these have been conclusively demonstrated in terms of pollutant exposure and immunotoxic effects in wildlife within individual experiments. Immunotoxicological biomarkers and biological monitoring tools are urgently needed to assess the extent of immunotoxicity in wildlife. Selection of universal biomarkers is hampered by the physiological diversity of immune systems in animals. However, by drawing on evidence from human epidemiology and tiered approaches in mammalian immunotoxicity evaluation, a selection of generic biomarkers of immunotoxicity in animals is suggested. Priorities for future research are also identified. PMID- 12739882 TI - Functional analysis of two matrix attachment region (MAR) elements in transgenic maize plants. AB - Matrix attachment regions (MARs) are binding sites for nuclear scaffold proteins in vitro, and are proposed to mediate the attachment of chromatin to the nuclear scaffold in vivo. Previous reports suggest that MAR elements may stabilize transgene expression. Here, we tested the effects of two maize MAR elements (P MAR from the P1-rr gene, and Adh1-MAR from the adh1 gene) on the expression of a gusA reporter gene driven by three different promoters: the maize p1 gene promoter, a wheat peroxidase (WP) gene promoter, or a synthetic promoter (Rsyn7). The inclusion of P-MAR or Adh1-MAR on P::GUS transgene constructs did not reduce variation in the levels of GUS activity among independent transformation events, nor among the progeny derived from each event. The Adh1-MAR element did not affect GUS expression driven by the WP promoter, but did modify the spatial pattern of expression of the Rsyn7::GUS transgene. These results indicate that, in transgenic maize plants, the effects of MAR elements can vary significantly depending upon the promoter used to drive the transgene. PMID- 12739883 TI - Renal tubule-specific expression and urinary secretion of human growth hormone: a kidney-based transgenic bioreactor growth. AB - Tissue-specific expression of human genes and secretion of human proteins into the body fluids in transgenic animals provides an important means of manufacturing large-quantity and high-quality pharmaceuticals. The present study demonstrates using transgenic mice that a 3.0 kb promoter of the mouse Tamm Horsfall protein (THP, or uromodulin) gene directs the specific expression of human growth hormone (hGH) gene in the kidney followed by the secretion of hGH protein into the urine. hGH expression was detected in renal tubules that actively produce the THP, that is, the ascending limb of Henle's loop and distal convoluted tubules. Up to 500 ng/ml of hGH was detected in the urine, and this level remained constant throughout the 10-month observation period. hGH was also detectable in the stomach epithelium and serum in two of the transgenic lines, suggesting position-dependent effects of the transgene and leakage of hGH from the site of synthesis into the bloodstream, respectively. These results indicate that the 3.0 kb mouse THP promoter is primarily kidney-specific and can be used to convert kidney into a bioreactor in transgenic animals to produce recombinant proteins. Given the capacity of urine production independent of age, sex and lactation, the ease of urinary protein purification, and the potentially distinct machinery for post-translational modifications in the kidney epithelial cells, the kidney-based transgenic bioreactor may offer unique opportunities for producing certain complex pharmaceuticals. PMID- 12739884 TI - Expression of rotavirus capsid protein VP6 in transgenic potato and its oral immunogenicity in mice. AB - Murine rotavirus gene six encoding the 41 kDa group specific capsid structural protein VP6 was stably inserted into the Solanum tuberosum genome by Agrobacterium tumefaciens mediated transformation. The molecular mass of plant synthesized VP6 capsid protein determined by immunoblot was similar to the size of both purified virus VP6 monomeric peptides and partially assembled virus-like particles. The amount of VP6 protein synthesized in transgenic potato leaf and tuber was determined by enzyme-linked immunosorbent assay to be approximately 0.01% of total soluble protein. Oral immunization of CD-1 mice with transformed potato tuber tissues containing VP6 capsid protein generated measurable titers of both anti-VP6 serum IgG and intestinal IgA antibodies. The presence of detectable humoral and intestinal antibody responses against the rotavirus capsid protein following mucosal immunization provides an optimistic basis for the development of edible plant vaccines against enteric viral pathogens. PMID- 12739885 TI - Induction of tolerance to a recombinant human enzyme, acid alpha-glucosidase, in enzyme deficient knockout mice. AB - When knockout mice are used to test the efficacy of recombinant human proteins, the animals often develop antibodies to the enzyme, precluding long-term pre clinical studies. This has been a problem with a number of models, for example, the evaluation of gene or enzyme replacement therapies in a knockout model of glycogen storage disease type II (GSDII; Pompe syndrome). In this disease, the lack of acid alpha-glucosidase (GAA) results in lysosomal accumulation of glycogen, particularly in skeletal and cardiac muscle. Here, we report that in a GAA-deficient mouse model of GSDII, low levels of transgene-encoded human GAA expressed in skeletal muscle or liver dramatically blunt or abolish the immune response to human recombinant protein. Of two low expression transgenic lines, only the liver-expressing line exhibited a profound GAA deficiency in skeletal muscle and heart indistinguishable from that in the original knockouts. The study suggests that the induction of tolerance in animal models of protein deficiencies could be achieved by restricting the expression of a gene of interest to a particular, carefully chosen tissue. PMID- 12739886 TI - A specific real-time quantitative PCR detection system for event MON810 in maize YieldGard based on the 3'-transgene integration sequence. AB - The increasing presence of transgenic plant derivatives in a wide range of animal and human consumables has provoked in western Europe a strong demand for appropriate detection methods to evaluate the existence of transgenic elements. Among the different techniques currently used, the real-time quantitative PCR is a powerful technology well adapted to the mandatory labeling requirements in the European Union (EU). The use of transgene flanking genomic sequences has recently been suggested as a means to avoid ambiguous results both in qualitative and quantitative PCR-based technologies. In this study we report the identification of genomic sequences adjacent to the 3'-integration site of event MON810 in transgenic maize. This genetically modified crop contains transgene sequences leading to ectopic expression of a synthetic CryIA(b) endotoxin which confers resistance to lepidopteran insects especially against the European corn borer. The characterization of the genome-transgene junction sequences by means of TAIL PCR has facilitated the design of a specific, sensitive and accurate quantification method based on TaqMan chemistry. Cloning of event MON810 3' junction region has also allowed to compare the suitability of plasmid target sequences versus genomic DNA obtained from certified reference materials (CRMs), to prepare standard calibration curves for quantification. PMID- 12739887 TI - Kidney-specific activity of the bovine uromodulin promoter. AB - A 10-kilobase (kb) lambda bacteriophage bovine genomic clone containing 5.4 kb of the 5'-flanking region, exons, and introns of bovine uromodulin gene was isolated. Transgenic mice containing 3.9 kb of the bovine uromodulin promoter and a lacZ reporter gene were generated by pronuclear microinjection. RT-PCR and northern blot analyses of transgene expression in various tissues of founder and F1 mice showed that the transgene was expressed exclusively in the kidney. In situ hybridization and histochemistry for lacZ demonstrated that transgene expression was restricted to tubule epithelial cells of the loop of Henle in the kidney. Stepwise 5' deletion analysis revealed that transfection of luciferase reporter constructs fused to various proximal 5'-flanking regions of the bovine uromodulin gene markedly increased luciferase activity in mouse renal epithelial cells but not in mesenchymal cells and that the most critical cis elements of the uromodulin gene are located within the 600 bp upstream region. PMID- 12739888 TI - Improved in planta expression of the human islet autoantigen glutamic acid decarboxylase (GAD65). AB - The smaller isoform of the enzyme glutamic acid decarboxylase (GAD65) is a major islet autoantigen in autoimmune type 1 diabetes mellitus (T1DM). Transgenic plants expressing human GAD65 (hGAD65) are a potential means of direct oral administration of the islet autoantigen in order to induce tolerance and prevent clinical onset of disease. We have previously reported the successful generation of transgenic tobacco and carrot that express immunoreactive, full-length hGAD65. In the present study, we tested the hypothesis that the expression levels of recombinant hGAD65 in transgenic plants can be increased by targeting the enzyme to the plant cell cytosol and by mediating expression through the potato virus X (PVX) vector. By substituting the NH2-terminal region of hGAD65 with a homologous region of rat GAD67, a chimeric GAD67(1-87)/GAD65(88-585) molecule was expressed in transgenic tobacco plants. Immunolocalization analysis showed that immunoreactive GAD67/65 was found in the plant cell cytosol. By using a radio immuno assay with human serum from a GAD65 autoantibody-positive T1DM patient, the highest expression level of the recombinant GAD67/65 protein was estimated to be 0.19% of the total soluble protein, compared to only 0.04% of wild-type hGAD65. Transient expression of wild-type, full-length hGAD65 in N. benthamiana mediated by PVX infection was associated with expression levels of immunoreactive protein as high as 2.2% of total soluble protein. This substantial improvement of the expression of hGAD65 in plants paves the way for immunoprevention studies of oral administration of GAD65-containing transgenic plant material in animal models of spontaneous autoimmune diabetes. PMID- 12739890 TI - Tritrophic interactions between transgenic potato expressing snowdrop lectin (GNA), an aphid pest (peach-potato aphid; Myzus persicae (Sulz.) and a beneficial predator (2-spot ladybird; Adalia bipunctata L.). AB - Tritrophic interactions between transgenic potato expressing the insecticidal lectin from snowdrop (Galanthus nivalis agglutinin; GNA), an aphid pest, Myzus persicae (Sulz.), and a beneficial predator, the 2-spot ladybird (Adalia bipunctata L.) were investigated. Clonal plants expressing GNA at 0.1-0.2% total soluble protein in leaves were used. No significant effects on development and survival of ladybird larvae fed on aphids from these transgenic plants were observed, with larval survival in the experimental group being 90% compared to 89% for controls. There were also no effects on subsequent female or male longevity. Female fecundity was also investigated. Although no significant differences (p > 0.05) were observed in egg production between control and experimental groups, a 10%, reduction (p < 0.01) in egg viability (determined by % hatch) occurred in ladybirds fed aphids reared on transgenic plants. Additional studies were carried out using aphids fed on artificial diet containing GNA, to deliver quantified levels of the protein to ladybird adults. GNA had no deleterious effects upon adult longevity, but resulted in a consistent trend for improved fecundity. Egg production was increased by up to 70% and egg viability also increased significantly. The results suggest that GNA is not deleterious to ladybirds. Results from these studies highlight the need to discriminate between direct and indirect effects when studying tritrophic interactions between plants/pests/natural enemies. Furthermore, it emphasises the importance of demonstrating 'cause and effect'. PMID- 12739889 TI - Analysis of cell-specificity and variegation of transgene expression driven by salmon prolactin promoter in stable lines of transgenic rainbow trout. AB - In order to identify the specificity and functionality of salmon prolactin (sPRL) promoter, transgenic rainbow trout carrying a construct comprising the 2.4 kb fragment of the 5' flanking region of Atlantic Chinook sPRL gene fused either to the reporter genes cat (sPRL-cat) or lacZ (sPRL-lacZ) were produced. sPRL-cat in transgenic F0 fish expressed strongly CAT only in the pituitary gland. Transgenic in F1-F4 lines harbouring sPRL-lacZ expressed beta-galactosidase (beta-gal) only in the follicular PRL-producing cells of the adenohypophysis. We observed heterocellular, mosaic distribution of beta-gal within PRL cell population and enormous variation of lacZ expression level between the littermates in the same transgenic line. Regardless of the transgene copy number, age or sex of transgenic fish, beta-gal expression was lactotroph-specific but variegated in all the nine F2 hemizygous lines analysed. One line harbouring a multicopy integration was followed up to F4 generation: the transgene was transmitted without modifications. Analysis of genomic DNA from pituitaries showed that lacZ sequences were highly methylated. LacZ expression was low and its transcripts, analysed by in situ hybridisation, showed a mosaic distribution within the pituitary gland. These data suggest that variegated expression of lacZ can occur at the transcription level owing to the silencing effect of lacZ gene. After proving the tissue-specific expression of reporter genes driven by the sPRL promoter, we tried to obtain the genetic ablation of PRL-producing cells,by transferring the same construct comprising diphtheria toxin DT-A gene (tox). However, the high mortality rate of sPRL-tox transformed embryos has embedded this study and no transgenic fish expressing tox were produced. The appropriateness of using transgenic strategies to analyse gene function in Salmonids is discussed, especially the implications of the multicopy integration patterns and of the variegated transgene expression. PMID- 12739891 TI - Transfer and expression of an artificial storage protein (ASP1) gene in cassava (Manihot esculenta Crantz). AB - In order to increase the nutritional quality of cassava storage roots, which contain up to 85% starch of their dry weight, but are deficient in protein, a synthetic ASP1 gene encoding a storage protein rich in essential amino acids (80%) was introduced into embryogenic suspensions of cassava via Agrobacterium mediated gene transfer. Transgenic plants were regenerated from suspension lines derived from hygromycin-resistant friable embryogenic callus lines. Molecular analysis showed the stable integration of asp1 in cassava genome and its expression at RNA level in transformed suspension lines. PCR and Southern analyses proved the transgenic nature of the regenerated plant lines. The expression of asp1 at RNA level was demonstrated by RT-PCR. The ASP1 tetramer could be detected in leaves as well as in primary roots of cultured transgenic plants by western blots. These results indicate that the nutritional improvement of cassava storage roots may be achieved by constitutive expression of asp1 in transgenic plants. PMID- 12739892 TI - Simple databases to monitor the generation and organisation of transgenic mouse colonies. AB - The generation and analysis of transgenic mice has become an important tool to progress our understanding of human and mouse gene function and its association with human genetic diseases. Animal models, based on genetically modified mice, both standard transgenic and knock-out animals, are increasingly being used worldwide. Monitoring of transgenic mouse production and transgenic mouse colonies is required to efficiently manage the resources that are available. Here, I describe three independent FileMaker databases (transgenics, mymouse and cages) that have been developed to track the generation of transgenic mice, the organisation of transgenic mouse colonies and the distribution of mice in cages. These three databases are freely available for academic use. PMID- 12739893 TI - Purification and biochemical characterization of a highly active cysteine protease ervatamin A from the latex of Ervatamia coronaria. AB - Ervatamia coronaria, a flowering plant (family Apocynaceae) indigenous to India, has medicinally important applications. A search for biochemical constituents of the latex of the plant yielded at least three thiol proteases with distinctly different properties. One of them, a highly active protease (ervatamin A), was purified to homogeneity by ion exchange and gel filtration chromatography. The enzyme exhibited high proteolytic activity toward natural substrates and amidolytic activity toward synthetic substrates. The pH and temperature optima for proteolytic activity were 8-8.5 and 50-55 degrees C, respectively. Proteolytic activity of the enzyme was strongly inhibited by thiol-specific inhibitors. The estimated molecular mass of the enzyme was 27.6 kDa. The extinction coefficient (epsilon(1%) 280) of the enzyme was estimated as 21.9, and the protein molecule consists of 8 tryptophan, 11 tyrosine and 7 cysteine residues. Isoelectric point of the purified enzyme was 8.37. Polyclonal antibodies raised against the pure enzyme gave a single precipitin line in Ouchterlony's double immunodiffusion and a typical color in ELISA. The N-terminal sequence of the enzyme showed conserved amino acid residues to other plant cysteine proteases. Ervatamin A shows high activity in relation to the other thiol proteases isolated from the same source. PMID- 12739894 TI - Morrenain b I, a papain-like endopeptidase from the latex of Morrenia brachystephana Griseb. (Asclepiadaceae). AB - A new cysteine endopeptidase (morrenain b I) has been purified and characterized from the latex of stems and petiols of Morrenia brachystephana Griseb. (Asclepiadaceae). Morrenain b I was the minor proteolytic component in the latex but showed higher specific activity than morrenain b II, which was the main active fraction. Both enzymes showed similar pH profiles and molecular masses, but kinetic parameters and N-terminal sequences were quite distinct, demonstrating that they are different enzymes instead of different forms of the same enzyme. PMID- 12739895 TI - Electrochemical evidence for the molten globule states of cytochrome c induced by N-alkyl sulfates at low concentrations. AB - The molten globule state (MG) of cytochrome c is the major intermediate of protein folding. The formation of MG state of cytochrome c is induced by n-alkyl sulfates such as sodium octyl sulfate (SOS), sodium dodecyl sulfate (SDS), and sodium tetradecyl sulfate (STS). The folding state of cytochrome c was monitored using circular dichroism (CD), isothermal titration calorimetry (ITC) and partial specific volumes. To explore a new approach for characterizing the MG conformation, cyclic voltametric studies of n-alkyl sulfates induced transition at acidic pH of cytochrome c (unfolded state, U) was carried out. Here, we have used a cystein-modified gold electrode, which is effective for direct rapid electron transfer to cytochrome c even in acid solutions, to directly observe electrochemistry in native (N) cytochrome c. Our results show that the extent of electron transfer is increased for U --> MG, and also the easiness of electron transferring occurred from MG --> N transition. Thus we demonstrate that the MG state of cytochrome c, induced by n-alkyl sulfates as salts with hydrophobic chains (hydrophobic salts), with different compactness reaches to near identical amount of electron transferring as N state. PMID- 12739896 TI - Spontaneous aggregation and cytotoxicity of the beta-amyloid Abeta1-40: a kinetic model. AB - The time dependency of the spontaneous aggregation of the fibrillogenic beta amyloid peptide, Abeta1-40, was measured by turbidity, circular dichroism, HPLC, and fluorescence polarization. The results by all methods were comparable and they were most consistent with a kinetic model where the peptide first slowly forms an activated monomeric derivative (AM), which is the only species able to initiate, by tetramerization, the formation of linear aggregates. The anti-Abeta antibody 6E10, raised against residues 1-17, at concentrations of 200-300 nM delayed significantly the aggregation of 50 microM amyloid peptide. The anti Abeta antibody 4G8, raised against residues 17-24, was much less active in that respect, while the antibody A162, raised against the C-terminal residues 39-43 of the full-length Abeta was totally inactive at those concentrations. Concomitant with the aggregation experiments, we also measured the time dependency of the Abeta1-40-induced toxicity toward SH-EPI cells and hippocampal neurons, evaluated by SYTOX Green fluorescence, lactate dehydrogenase release, and activation of caspases. The extent of cell damage measured by all methods reached a maximum at the same time and this maximum coincided with that of the concentration of AM. According to the kinetic scheme, the latter is the only transient peptide species whose concentration passes through a maximum. Thus, it appears that the toxic species of Abeta1-40 is most likely the same transient activated monomer that is responsible for the nucleation of fibril formation. These conclusions should provide a structural basis for understanding the toxicity of Abeta1-40 in vitro and possibly in vivo. PMID- 12739898 TI - Stability studies on a lipase from Bacillus subtilis in guanidinium chloride. AB - Lipase from Bacillus subtilis is a "lidless" lipase that does not show interfacial activation. Due to exposure of the active site to solvent, the lipase tends to aggregate. We have investigated the solution properties and unfolding of the lipase in guanidinium chloride (GdmCl) to understand its aggregation behavior and stability. Dynamic light scattering (DLS), near- and far-UV circular dichroism, activity and intrinsic fluorescence of lipase suggest that the protein undergoes unfolding between 1 M and 2 M GdmCl. The polarity sensitive dye, 1,1', bis-(4anilino)naphthalene-5,5"-disulfonic acid (bis-ANS), a probe for hydrophobic pockets, binds cooperatively to the native lipase. An intermediate populated in 1.75 M GdmCl that strongly binds bis-ANS was identified. Tendency of the native protein to aggregate in solution and specific binding to bis-ANS confirms that the lipase has exposed hydrophobic pockets and this surface hydrophobicity strongly influences the unfolding pathway of the lipase in GdmCl. PMID- 12739897 TI - Chicken deoxyribonuclease: purification, characterization, gene cloning and gene expression. AB - Chicken DNase was purified to apparent homogeneity from the pancreas extract. It showed two isoforms, A and B forms, on cation-exchange chromatography. On SDS PAGE it was a 30-kDa protein. When analyzed on an electrospray-mass analyzer, form A showed a major mass peak of 30859, and form B, 30882. The enzyme was bound to concanavalin A, indicating its glycoprotein nature. The carbohydrate side chain could be removed by endoglycosidase F. Chicken DNase was activated by metal ions and for half-maximum activation, Mn2+ and Mg2+ required were 1 mM and 4 mM, respectively. The pH optimum was between 7 and 8 depending on the metal ions used. In the presence of Cu2+, it was almost completely inactivated by 0.1 M iodoacetate within 1 min. In the absence of Ca2+ at pH 8, chicken DNase resisted to the trypsin or beta-mercaptoethenol inactivation. When the purified enzyme was subjected to protein sequencing, approximately 93% of the sequence was established. Based on the amino acid sequence, the cDNA of chicken DNase was amplified, cloned and sequenced. The cDNA sequence consisted of 1079 nucleotides in which 67 were of the 5'-untranslated region and 166 of the 3' and, in the 5' untranslated region, two types of sequences occurred. The polypeptide chain of 282 amino acids, translated from the open reading frame, was composed of the mature protein of 262 amino acids and a putative signal peptide of 20 amino acids. As compared with mammalian DNases, chicken DNase had an overall 58 +/- 61% sequence identity, one less potential N-glycosylation site, and one extra disulfide. The cDNA was cloned into the pET15b expression vector. When induced, active recombinant chicken DNase was expressed in Escherichia coli strain BL21(DE3)pLysS and was present in the insoluble fraction of cell lysates. PMID- 12739899 TI - Purification, electrophoretic behavior, and kinetics of iron release of liver ferritin of Dasyatis akajei. AB - From the liver of fish Dasyatis akajei, ferritin has been isolated by thermal denaturation and ammonium sulfate fractionation and then further purified by anion exchange chromatography and gel exclusion chromatography. The molecular weight of the liver ferritin of D. akajei (DALF) was measured to be 400 kDa by PAGE. Moreover, SDS-PAGE experimentation indicates that protein shell of DALF consists of the H and L subunits with molecular weight of 18 and 13 kDa, respectively. Using isoelectric focusing with pH ranging from 5.0 to 6.0, the ferritin purified by the PAGE exhibited three bands with different pI values in the gel slab. Diameters of the protein shell and iron core were also investigated by transmission electron microscope and determined to be 10-12 nm and 5-8 nm, respectively. A kinetic study of DALF reveals that the rate of self-regulation of the protein shell rather than the complex surface of the iron core plays an important role in forming a process for iron release with mixed orders. PMID- 12739900 TI - E292 is important for the aminoacylation activity of Escherichia coli leucyl-tRNA synthetase. AB - Escherichia coli leucyl-tRNA synthetase (LeuRS) has a large connecting polypeptide (CP1) inserted into its active site. It was demonstrated that the peptide bond between E292-A293 was crucial for the aminoacylation activity of E. coli LeuRS. To investigate the effect of E292 on the function of Escherichia coli LeuRS, E292 was mutated to K, F, S, D, Q and A. These mutations at 292 did not change the specific activity of the amino acid activation reaction. Though the conformational change of these mutants was not detected in CD, their aminoacylation activities were impaired to varying extents. The mutation of E to K decreased the aminoacylation activity to the largest extent. Analysis of the Km values of these mutants for the three substrates showed that the E292 was not involved in the binding of leucine and that all mutants had stronger binding with ATP. PMID- 12739901 TI - Molecular cloning and structural characterization of the hagfish proteinase inhibitor of the alpha-2-macroglobulin family. AB - The "most primitive" living vertebrate the hagfish has a dimeric proteinase inhibitor, a protein homologous to human alpha2-macroglobulin, in its plasma at high concentration. Although the hagfish proteinase inhibitor has been isolated and its function and quaternary structure studied, its primary structure, subunit composition and fragmentation process remain unclear. In this study, hagfish proteinase inhibitor cDNA was cloned, sequenced and cDNA-deduced amino acid sequence was analyzed. A large fraction of homosubunits in the dimeric structure of the protein has undergone a cleavage at a specific arginyl residue (Arg833) while the rest retained their chain integrity without being processed. Thus random combinations of processed and nonprocessed subunits in the dimeric structure of this protein result in different molecular conformers and generate a complicated multiband pattern in SDS-PAGE. It was further demonstrated by proteolytic analysis that the hagfish inhibitor has no susceptible arginyl residues within its bait region and thus incapable of trapping arginine specific proteinases. This implies that the specific subunit cleavage at Arg833 was caused by an unknown arginine specific proteinase which escaped from the entrapment by the hagfish inhibitor. PMID- 12739902 TI - Molecular characterization of GroES and GroEL homologues from Clostridium botulinum. AB - We report novel findings of significant amounts of 60- and 10-kDa proteins on SDS PAGE in a culture supernatant of the Clostridium botulinum type D strain 4947 (D 4947). The N-terminal amino acid sequences of the purified proteins were closely related to those of other bacterial GroEL and GroES proteins, and both positively cross-reacted with Escherichia coli GroEL and GroES antibodies. Native GroEL homologue as an oligomeric complex is a weak ATPase whose activity is inhibited by the presence of GroES homologue. The 2634-bp groESL operon of D-4947 was isolated by PCR and sequenced. The sequence included two complete open reading frames (282 and 1629 bp), which were homologous to the groES and groEL gene family of bacterial proteins. Southern and Northern blot analyses indicate that the groESL operon is encoded on the genomic DNA of D-4947 as a single copy, and not on that of its specific toxin-converting phage. PMID- 12739903 TI - Cognitive rehabilitation with patients having persistent, severe psychiatric disabilities. AB - The effectiveness of cognitive rehabilitation in ameliorating the symptomatic, cognitive, and functional deficits associated with schizophrenia and schizoaffective disorders was assessed in this prospective study. Thirty-eight participants who met DSM-IV criteria were assigned to cognitive rehabilitation treatment or treatment as usual (TAU) groups, using the method of minimization to equalize groups on prognostic variables believed to affect outcome (i.e., duration and severity of illness, Clozapine). Participants were assessed at baseline, treatment end, and 3-month follow-up. Improvement across time was found for both groups in delayed visuospatial memory and visual information processing speed, and the participant's status on the prognostic variables was found to be related to level of performance on measures of delayed visuospatial memory, negative symptoms, and speech disturbance. However, the findings did not provide evidence that cognitive rehabilitation is associated with greater improvement than TAU. Nor did the findings indicate that prognosis interacted with treatment to produce differential treatment outcomes. PMID- 12739904 TI - Crime among homeless military veterans who abuse substances. AB - This was a study of crime among 188 homeless persons who were in a Veterans' Affairs Medical Center program for substance abusers. The purpose of the study was to find out: (a) what proportion committed crimes, and (b) what other problems, relational factors, and personal attributes predict crime. Data indicate 27% of these homeless veterans committed nuisance offenses, and 41% have committed crimes in the past year. Logistic regression procedures indicated that alcohol and other drug abuse, less education, lack of employment, psychiatric problems, and living with a substance abuser increased the odds of committing crimes. This study also finds that physical and sexual abuse before 18 years of age increases the odds of committing crimes, whereas self-efficacy, ego integrity, and resilience decrease these odds. These latter factors have received scant attention in the literature on homeless substance abusers, and yet they are among the strongest predictors of crime. Implications for psychiatric rehabilitation are discussed. PMID- 12739905 TI - Towards an integrated, structural model of psychiatric rehabilitation. AB - In the past two decades, the number of principles and approaches that have been touted as psychiatric rehabilitation have overwhelmed the service provider. As a result, the field seems to be bombarded by contradictory ideas. In this paper, I seek to resolve this problem by providing an overview of psychiatric rehabilitation in terms of four key structures: goals (the reasons why people with psychiatric disabilities seek rehabilitation), strategies (the tools which service providers might use to help consumers reach their goals), settings (the places in which providers and consumers use these tools), and roles (the variety of professional and personal backgrounds which influence providers in carrying out their duties). In this paper, I review the research literature that has identified relevant examples of each of these structural dimensions. This description of psychiatric rehabilitation is based on a two-factor model of the life challenges that arise from mental illness: the disabilities produced by the disease and the loss of opportunity that result from stigma and discrimination. The paper also includes a brief discussion of the two-factor model. PMID- 12739906 TI - Transforming sheltered workshops into affirmative businesses: an outcome evaluation. AB - This paper examines the outcomes associated with Voices, Opportunities and Choices Employment Club (VOCEC), which used the resources of a sheltered workshop to evolve affirmative businesses for people receiving mental health services. Quantitative and qualitative methodologies were used to understand the impact of VOCEC on employment and well-being outcomes compared to the former sheltered workshops and to competitive work. The results suggest that VOCEC has been successful in creating paid employment opportunities at levels approximating minimum wage and establishing a structure to incubate new businesses. The evaluation revealed the process by which Associates experience their participation as rewarding. It also indicated constraints on participation within the VOCEC model. PMID- 12739907 TI - Discovering recovery. AB - This paper describes the interaction between a micro-history of recovery and a personal recovery history. It begins with a narrative of the author's own recovery experience as he participates in a series of recovery initiatives in a Midwestern state. The impact of these history-graded experiences on his human life-span development is then examined in the context of recovery. Lessons learned are discussed, and some conclusions are drawn. Implications for mental health professionals and people receiving mental health services are also explored. The benefits of getting involved are highlighted, as well as some concerns. PMID- 12739908 TI - "It's like being in another world": demonstrating the benefits of therapeutic horseback riding for individuals with psychiatric disability. AB - Five adults with longstanding histories of psychiatric disabilities were recruited for a ten-week therapeutic horseback riding program. Individuals learned basic riding skills and had the opportunity to bond with a horse. In addition, the riders participated in a post-riding process group that used artistic and creative exercises to promote individual expression. By the end often weeks, the riders reported success in learning basic horsemanship and, in doing so, also reported additional psychosocial benefits, including an augmented sense of self-efficacy and self-esteem. In sum, this adjunctive therapy can facilitate the recovery process. PMID- 12739909 TI - The New Hampshire financing policy. AB - OBJECTIVE: This study sought to describe the New Hampshire incentive financing policy and report on its results over the first seven years. METHODS: The description of the policy was based on reviewing state documents and interviews with state officials. The data were obtained from state records. RESULTS: On the targeted outcomes of employment and psychiatric hospitalization the statewide improvement was dramatic. The service system concurrently became more efficient. CONCLUSION: The results suggest the incentive financing based on client outcomes may be a potent state strategy for improving service system performance. PMID- 12739910 TI - Social network correlates among people with psychiatric disabilities. AB - This study explores three social network characteristics as they relate to demographic, clinical, and quality of life factors. The sample consists of 219 participants with serious mental illness. Results indicate that clients' social networks were small, and that network size is robustly related to symptoms and other measures of clinical and cognitive functioning, as well as with quality of life and self-esteem. The expected relation between network density and either the demographic variables or quality of life was not found. Moderate levels of network density, however, were associated with fewer psychiatric symptoms and higher IQ. Network multiplexity was most highly associated with demographic variables, such as ethnic minority status, less education, never being married, and living with family. PMID- 12739911 TI - Connection and autonomy in the case management relationship. AB - This study examines the effects of connection and autonomy in the client-case manager relationship on treatment participation, satisfaction with case management, and satisfaction with social life. Three-month case manager ratings of connection were positively correlated with 9-month treatment participation. Six-month client and case manager ratings of connection were positively correlated with improvements in all three outcomes at nine months. Within this time frame, autonomy was positively correlated with treatment participation and satisfaction with case management and negatively correlated with satisfaction with social life. These findings suggest that balancing connection and autonomy in different ways is important in the case management relationship. PMID- 12739912 TI - Another way of avoiding the "single model trap". AB - Another way to end the "model wars" (Hughes & Clement, 1999; IAPSRS, 1998) is to take what works best from different models and blend them. At two different sites, practitioners being trained to deliver ACT received additional training in the psychiatric rehabilitation and case management technology that had been previously developed at Boston University. Feedback from the practitioners indicated that the blending of these two "models" was helpful and additive. A 4 year period of data collection at one site using a simple pre-post test design showed inpatient days were reduced by about 90% for 80 individuals who were considered to be high utilizers of inpatient services. It appears that the integration of separately developed model approaches bears further study. PMID- 12739913 TI - Computer experience and training interests of psychosocial rehabilitation program participants. AB - The authors report data from the first large-scale attempt (N = 262) to examine computer use among people with a psychiatric disability and interest in computer training. A large number of respondents have used a computer (71%) for a number of activities and approximately 56% were "very" or "quite" interested in computer training. Previous work experience, interest in a job involving computers, and a belief that training would be helpful in obtaining such a job were good predictors of interest in computer training. The results suggest that there is sufficient interest in computer training to justify the development of supported education in this area. PMID- 12739914 TI - Interpreting narratives of motivation and schizophrenia: a biopsychosocial understanding. AB - BACKGROUND: The concept of motivation involves a complex interplay of biopsychosocial and environmental determinants. For individuals diagnosed with schizophrenia, motivation has traditionally been approached from a neuro biological standpoint, obscuring this complexity. The findings from this study underscore the importance of broadening our understanding of motivation and schizophrenia through an exploration of individual perspectives and identification of the psychosocial factors that clarify the experience of diminished motivation. PMID- 12739915 TI - Recovery: always a work in progress. AB - Work can be an important part of recovery. It is my belief that my work as a Patient Advocate at Norristown State Hospital has contributed greatly to my stability. This article discusses some of the contributing factors to that stability. PMID- 12739916 TI - Herbal medicines for children. PMID- 12739918 TI - Considering culture, complementary medicine, and spirituality in pediatrics. AB - Whether or not it is apparent, cultural and spiritual diversity exist in many clinical encounters, both with patients and with colleagues. These issues may cloud other concerns, contribute to inadequate or misleading communication, and affect lifestyle and therapeutic choices. Given the rapid rise in complementary therapies and the growing diversity in North America, the challenges of providing sensitive, compassionate, comprehensive care are compounded. Reflecting on the ways cultural backgrounds, religious beliefs and complementary medicine affect physicians, colleagues and patients may enrich, clarify, deepen, and improve the effectiveness of our clinical practice. The cases presented here offer an opportunity for role playing and reflection to support this process. PMID- 12739919 TI - Effect of oral supplementation on catch-up growth in picky eaters. AB - Ninety-two subjects ages 36 to 60 months who had picky-eater behavior and evidence of growth faltering were randomized to receive either nutrition counseling alone, or nutrition counseling plus a nutritional supplement (Study) for 90 days. The Study group had significantly greater increases in weight and height. There were no significant differences between groups in changes in appetite or activity levels, or in gastrointestinal symptom scores. The percent of subjects who developed upper respiratory tract infections was significantly lower in the Study group. These data suggest that a nutritional supplement in addition to nutrition counseling promote catch-up growth and may contribute to lower rates of infectious disease in children with picky eater behavior. PMID- 12739917 TI - Working with challenging young children: relations between child temperament, response to novelty and sensory processing. PMID- 12739920 TI - Efficacy of penicillin vs. amoxicillin in children with group A beta hemolytic streptococcal tonsillopharyngitis. AB - The purpose of this study was to compare the bacteriologic and clinical efficacy of oral penicillin versus amoxicillin as first-line therapy for group A beta hemolytic streptococcal (GABHS) tonsillopharyngitis. The prospective observational study was conducted over 18 months (January 2000-June 2001). Children enrolled had acute onset of symptoms and signs and a laboratory documented GABHS tonsillopharyngitis illness. Follow-up examination and laboratory testing occurred 10 +/- 4 days following completion of treatment. In total, 389 patients were enrolled (intent-to-treat group): 195 received penicillin V and 194 received amoxicillin. Fifty-six of the penicillin-treated and 57 amoxicillin-treated patients refused to take the drug, or were noncompliant, or did not return for the follow-up visit, leaving 276 patients in the per-protocol group: 139 penicillin-treated and 137 amoxicillin-treated. Bacteriologic cure for amoxicillin-treated children occurred in 76% versus 64% in the penicillin-treated children (p = 0.04). The clinical cure rate for amoxicillin-treated children was 84% compared to 73% in the penicillin-treated children (p = 0.03). Since treatment allocation was not randomized, logistic regression analysis was used to adjust for treatment group differences. The odds ratio (OR) estimate for cure for patients in the amoxicillin versus penicillin V treatment group remained significant (OR = 1.84, 95% confidence interval 1.02 3.29); the same was true for dinical cure (OR = 1.99, 95% CI = 1.02-3.87). Amoxicillin may be superior to penicillin for bacteriologic and clinical cure of GABHS tonsillopharyngitis. PMID- 12739921 TI - Pediatric pain measurement using a visual analogue scale: a comparison of two teaching methods. AB - The goals of this study were to evaluate the validity of the visual analogue scale (VAS) for young children and to compare a newly developed method of teaching children to use a VAS with one used in our previous studies. It was hypothesized that the new method would increase the number of children who understand the VAS and correctly mark their responses on the VAS line. The association between child's age and ability to understand the VAS was also evaluated. One hundred-six children with a laceration requiring sutures and receiving a lidocaine injection for local anesthesia participated in the study. They ranged in age from 5 to 14 years. Two outcome measures were used to assess the baseline and lidocaine injection pain: a 5-point Likert scale and a VAS. A calibration study was used to determine whether the subjects were able to use the VAS to make proportional judgments about their perceptions. Teaching method had no effect on the number of subjects who could correctly mark their responses on the VAS line, nor did it significantly increase the number of subjects who could understand the concept of the VAS. Subjects who were able to understand the VAS were significantly older (mean = 9.8 years, SD = 2.8) than those who did not (mean = 8.2 years, SD = 2.5). Overall, only about one third of the subjects were able to correctly mark the VAS and understand the concept of the VAS. Other measures of pain that are better understood by young children may be more valid indicators of pain than the VAS. PMID- 12739922 TI - Survey of physician attitudes and practices related to pediatric obesity. AB - The purpose of this study was to survey physicians regarding their attitudes and practices related to the treatment of pediatric obesity in a primary care setting. Surveys were sent to physicians who were members of the American Academy of Pediatrics and the American Academy of Family Physicians practicing in the Southern New England area (Connecticut, Massachusetts, and Rhode Island). The 14 item survey consisted of three main areas of focus: attitudes toward obesity, treatment and referral approaches, and barriers to addressing weight concerns in children and adolescents. Physicians estimated that 27.7% of their adolescent and 23% of their child patients are overweight. The frequency with which physicians address weight issues with both child and adolescent patients appears to increase incrementally with the patient's level of overweight. When addressing obesity, one fourth of physicians think that they are not at all or only slightly competent, while 20% report feeling not at all or only slightly comfortable. These findings suggest that physicians would benefit from additional training and education regarding safe and efficacious intervention strategies for pediatric obesity, to effectively integrate the discussion of weight issues into the primary care setting. PMID- 12739923 TI - The usefulness of preoperative screening for bleeding disorders. PMID- 12739924 TI - Congenital cystic lung disease: diagnostic and therapeutic considerations. AB - Congenital lung cysts such as congenital cystic adenomatoid malformation, pulmonary sequestration, congenital lobar emphysema, and bronchogenic cysts are rare but fascinating anomalies of lung development. While there are many similarities in terms of their presenting features, there are particular differences between the diagnostic groups that are important to highlight, especially in relationship to approaches to imaging and long-term outcome. A case of each entity is presented with an emphasis on the contemporary approach to diagnostic investigations and therapeutic options. PMID- 12739925 TI - Kawasaki disease in very young infants: high prevalence of atypical presentation and coronary arteritis. PMID- 12739926 TI - Early intravenous gamma globulin retreatment for refractory Kawasaki disease. PMID- 12739928 TI - Pediatric subspecialty consultation in a university hospital: a reappraisal. PMID- 12739927 TI - Curing bedwetting on the spot: a review of one-session cures. PMID- 12739929 TI - Moyamoya disease in a four-month-old: a case study. PMID- 12739930 TI - Hydroformylation of terminal alkenes supported on solid phase: synthetic tool for combinatorial chemistry. PMID- 12739932 TI - Attachment of unreactive amines to the solid support: synthesis of phenyl substituted anilines, 2-aminopyridines, and 2-aminopyrimidines. PMID- 12739931 TI - Modular liquid-phase parallel synthesis of a highly diverse ligand library. PMID- 12739933 TI - An efficient, stereoselective solid-phase synthesis of beta-lactams using Mukaiyama's salt for the Staudinger reaction. PMID- 12739934 TI - Parallel synthesis of antimalarial rhodacyanine dyes by the combination of three components in one pot. PMID- 12739935 TI - Two-photon microscopy to spatially resolve and quantify fluorophores in single bead chemistry. PMID- 12739937 TI - High-performance thin-layer chromatography method for assessment of the quality of combinatorial libraries, and comparison with liquid chromatography-ultraviolet mass spectrometry. AB - A high-performance thin-layer chromatography (HPTLC) method was developed for fast evaluation of the purity of solid-phase synthesis products. The results obtained were in good agreement with results obtained by the LC-MS method (r(2) = 0.8404) or by the LC-UV method (r(2) = 0.8053), confirming the suitability of HPTLC for purity analysis of combinatorial syntheses. The synthesis products can be quantified and identified by measuring UV densitograms or in situ UV spectra or by ESI-MS after isolation of the zone of interest. A new, simple, and fast method for transferring the zone of the analyte from the plate to the ESI-MS equipment is described. The new HPTLC method enables rapid and efficient analysis of approximately 40 samples in parallel. As such, it offers a cheaper and easier way to analyze the purity of synthesis products than the commonly used LC-UV-MS. PMID- 12739936 TI - An improved method for rapid sequencing of support-bound peptides by partial edman degradation and mass spectrometry. PMID- 12739938 TI - Ligand-based combinatorial design of selective purinergic receptor (A2A) antagonists using self-organizing maps. AB - A virtual screening procedure based on a topological pharmacophore similarity metric and self-organizing maps (SOM) was developed and applied to optimizing combinatorial products functioning as P(1) purinergic receptor antagonists. The target was the human A(2A) receptor. A SOM was developed using a set of biologically tested molecules to establish a preliminary structure-activity relationship. A combinatorial library design was performed by projecting virtually assembled new molecules onto the SOM. A small focused library of 17 selected combinatorial products was synthesized and tested. On average, the designed structures yielded a 3-fold smaller binding constant ( approximately 33 vs approximately 100 nM) and 3.5-fold higher selectivity (50 vs 14) than the initial library. The most selective compound obtained revealed a 121-fold relative selectivity for A(2A) with K(i) (A(2A)) = 2.4 nM, and K(i) (A(1)) = 292 nM. This result demonstrates that it was possible to design a small, activity enriched focused library with an improved property profile using the SOM virtual screening approach. The strategy might be particularly useful in projects in which structure-based design cannot be applied because of a lack of receptor structure information, for example, in the many projects aiming at finding new GPCR modulators. PMID- 12739939 TI - High-throughput analysis in catalysis research using novel approaches to transmission infrared spectroscopy. AB - This study has demonstrated that high-throughput FTIR transmission measurements using a newly designed array-based support formed using silicon wells and a silicon wafer is a very useful and robust tool for the characterization of polymer composition for combinatorial materials research. The comonomer content in copolymers can be measured accurately with a fully automated throughput of >300 samples/day (8 h). The transmission measurement is more robust, reliable, and easier to automate than other spectroscopic methods. The support itself provides excellent resistance to aggressive organic solvents at elevated temperatures and allows the unattended deposition and preparation of polymer films for infrared analysis. Because of the excellent durability of the support with respect to the solvent, the support can be rinsed and reused many times. This high-throughput approach to infrared transmission spectroscopy can be used for measuring a wide array of polymer characteristics: vinyl content, geometrical isomers, crystallinity, and tacticity. As well, this IR approach can be used to predict the oxidative stability of the antioxidant packages. Because the support provides a means of containing hot polymer solutions while the solvent evaporates, the support is also suitable for high-throughput nanoindentation methods for the determination of modulus and other physical properties of the polymer. PMID- 12739941 TI - Solid-phase synthesis of 7-acylamino-1,4-benzodiazepine-2,5-diones. AB - A method for the synthesis of polymer-bound 7-acylamino-benzodiazepine-2,5-diones is described. The amino group of an alpha-amino acid is linked to polystyrene or TentaGel resin via reductive amination of polymer-bound 4-alkoxy-2,6 dimethoxybenzaldehyde. Acylation with unprotected 5-nitroanthranilic acid is followed by base-catalyzed ring closure. Reduction of the nitro group yields enantiomerically pure 7-aminobenzodiazepin-2,5-dione attached via the N-4 atom to the resin. Acylation of the amino group on the aromatic ring with acid chlorides in N-methylpyrrolidone (no DMF, no base!) followed by cleavage from the resin using TFA/Me(2)S/water (90:5:5) provides the acylated benzodiazepinones in 52-69% (PS resin) and 41-48% (TG resin) yield (based on the theoretical loading) and >70% purity (HPLC, 210 nm). Using Fmoc-protected tyrosine fluoride in NMP gives the amino acid-coupled benzodiazepinones in 24% (PS resin) and 31% (TG resin) yield. PMID- 12739940 TI - Micro-X-ray fluorescence as a general high-throughput screening method for catalyst discovery and small molecule recognition. AB - A powerful high-throughput screening technique is described for the rapid screening of bead-based libraries for catalyst discovery and molecular recognition. Micro-X-ray fluorescence (MXRF) screens materials for elemental composition with mesoscale analysis. This method is nondestructive and requires minimal sample preparation and no special tags for analysis, and the screening time is dependent on the desired sensitivity. The speed, sensitivity, and simplicity of MXRF as a high-throughput screening technique were applied to screen bead-based libraries of oligopeptides for phosphate hydrolysis catalysts and molecular recognition of selective receptors for the degradation products and analogues of chemical warfare agents. This paper demonstrates the analytical or HTS capability of MXRF for combinatorial screening. It is meant only to show the capabilities of MXRF and is not meant as an exhaustive study of the catalyst and molecular recognition systems presented. PMID- 12739942 TI - The solid-phase synthesis and use of N-monosubstituted piperazines in chemical library synthesis. AB - An efficient solid-phase synthesis of mono-N-substituted piperazines is presented. The key transformation involves a selective borane amide bond reduction in the presence of a carbamate resin linkage. This synthetic route takes advantage of the large diverse pool of commercially available carboxylic acids, acid chlorides, and sulfonyl chlorides. The solid-phase approach facilitates parallel processing by eliminating the need for column chromatography after each synthetic step. The N-monosubstituted piperazines were shown to react with polymeric activated tetrafluorophenol (TFP) reagents to generate arrays of amides and sulfonamides in good purity for biological testing. PMID- 12739943 TI - Suzuki cross-coupling of solid-supported chloropyrimidines with arylboronic acids. AB - The utility of the Suzuki cross-coupling to synthesize biaryl compounds is expanded herein to include reactions of resin-supported chloropyrimidines with boronic acids. In particular, an efficient method is described for the synthesis of a library of biaryl compounds from solid-supported chloropyrimidines. The Suzuki reaction was performed in an inert atmosphere using Pd(2)(dba)(3)/P(t Bu)(3) as catalyst, spray-dried KF as base, and THF as solvent. The reaction was allowed to proceed overnight at 50 degrees C. Upon cleavage with acid, a library of 4-(substituted amino)-6-arylpyrimidines was obtained in moderate yield and high purity. PMID- 12739944 TI - A facile solid-phase synthesis of substituted 2(5H)-furanones from polymer supported alpha-selenocarboxylic acids. AB - The synthesis of polystyrene-supported alpha-selenoacetic acid, alpha selenopropionic acid, and alpha-selenophenylacetic acid is described. The reaction of the dilithio derivatives of polymer-supported alpha-selenocarboxylic acids with racemic epoxides or optically active styrene oxide afforded polystyrene-supported gamma-substituted alpha-selenobutyrolactones. The alpha alkylation reaction of gamma-substituted polystyrene-supported alpha selenobutyrolactones provided another route for the synthesis of polystyrene supported alpha,gamma-disubstituted alpha-selenobutyrolactones. Subsequent oxidation-elimination with an excess of 30% hydrogen peroxide at room temperature afforded substituted (3- and 5-mono-; 3,4- and 3,5-di) 2(5H)-furanones in high yields and good purities. PMID- 12739946 TI - Amino acid-derived heterocycles as combinatorial library targets: spirocyclic ketal lactones. AB - The spirocyclic ketal-lactone frameworks of 3 and 4 were designed as novel structures amenable to combinatorial synthesis. The synthesis of representative analogues was developed in solution and on solid support, the scope of effective input materials was determined, and the stability and stereochemistry of the products was evaluated. The spirocycles are obtained in modest overall yields (5 36%) and excellent purities (>72%) and offer a promising motif for combinatorial prospecting libraries. PMID- 12739945 TI - Amino acid-derived heterocycles as combinatorial library targets: bicyclic aminal lactones. AB - The incorporation of alpha-amino acids into heterocyclic structures is an effective strategy for generating peptidomimetics and combinatorial library scaffolds. This report describes the synthesis of novel bicyclic aminal lactones 3 by base-catalyzed cyclization of N-(2-oxoalkyl)-dipeptide esters 8. Assembly of the acyclic precursor 8 can be carried out on solid phase, with variation at four positions; cyclative release ensures high product purity in the final step. Cyclization affords the exo isomer stereospecifically when one chiral center is present in the precursor, or when both amino acids have the same configuration. PMID- 12739947 TI - Solid-phase synthesis of nucleoside analogues. AB - The synthesis of a 25 000 member library of nucleoside analogues as discrete compounds in milligram quantities is described. The use of the Nanokan technology developed by IRORI (Discovery Partners International) together with macroporous solid support allowed us to develop a highly reliable and practical synthetic route for the high-throughput derivatization of both the pyrimidine and purine nucleoside scaffold. A 2',3'-acetal linkage of the scaffolds to the solid support proved to be stable enough for the chemical transformations employed, yet labile enough for mild cleavage conditions to yield final products in high purity. The publication represents an example for combining synthetic organic chemistry on advanced scaffolds with the latest technologies of combinatorial chemistry in order to provide both industrial and academic institutions with compounds in high number and quality, thereby accelerating the search for novel biological targets and drug development. PMID- 12739948 TI - Solution- and solid-phase parallel synthesis of 4-alkoxy-substituted pyrimidines with high molecular diversity. AB - A simple and straightforward methodology toward the synthesis of novel 2,6 disubstituted-4-alkoxypyrimidine derivatives of type 16 and 19 has been developed. This methodology, initially developed in solution, can be perfectly adapted to the solid support under analogous conditions, taking full advantage of automated parallel synthesis systems. This successful methodology benefits from the key role played by the thioether linkage placed at the 2-position in 3, 9, or 13 in a double manner: on one side, the steric effect exerted by the thioether linkage is likely to be responsible for the very high observed selectivity toward the formation of the O-alkylation products. On the other side, this sulfur linkage can serve not only as a robust point of attachment for the heterocycle, stable to a number of reaction conditions, but also as a means of introducing a new element of diversity through activation to the corresponding sulfone (safety catch linker concept) and subsequent ipso-substitution reaction with a variety of different N-nucleophiles. PMID- 12739949 TI - Development of a custom high-throughput preparative liquid chromatography/mass spectrometer platform for the preparative purification and analytical analysis of compound libraries. AB - Solution-phase parallel synthesis has had a profound impact on the speed of compound synthesis delivering relatively pure compounds (>80%) in short order. However, to develop structure activity relationships (SAR) for a compound series, each library member should preferably be >95% pure. Historically, achieving and quantifying such high-purity criteria for each library member proved to be the slow step for most lead discovery groups. To address this issue, significant modifications have been made to a commercial Agilent preparative LC/MS system to allow for the general mass-guided purification of diverse compound libraries. The custom modifications include (1) the "DMSO slug" approach for the purification of samples with poor solubility; (2) an active splitter to reduce system back pressure, reduce the delay volume, and allow for a variable split ratio; (3) a sample loading pump for the quick purification of large, dilute samples; (4) a preparative column-selection valve to quickly change column selectivity or sample loading; and (5) an analytical injector with a separate flow path for crude reaction or fraction analyses. PMID- 12739950 TI - Nitrophenol resins for facile amide and sulfonamide library synthesis. AB - Novel nitrophenol solid supports based on various resin materials (polystyrene, TentaGel, macroporous, PEGA, and silica gel) are reported for facile amide and sulfonamide library synthesis. The broad choice of resin materials available will allow the reaction to occur successfully in solvents ranging from nonpolar organic solvents to aqueous media. PMID- 12739952 TI - Structural and catalytic properties of bis(guanidine)copper(I) halides. AB - The neutral, bicyclic guanidine 1,3,4,6,7,8-hexahydro-2H-pyrimido[1,2 a]pyrimidine (hppH) coordinates to Cu(I) halides via the imine nitrogen, and the complexes are stabilized by intramolecular NH...X interactions (X = Cl, Br) or are partially dissociated (X = I) in the solid state; in all cases, fluxional behavior is observed in solution. Selected complexes have been tested as catalysts for the polymerization of methyl methacrylate. PMID- 12739951 TI - Rapid parallel synthesis of dipeptide diphenyl phosphonate esters as inhibitors of dipeptidyl peptidases. AB - In this paper, we present a parallel synthesis of several series of dipeptide diphenyl phosphonates that are known to be irreversible inhibitors of serine proteases. Polymer-assisted solution-phase synthesis (PASP) is used for the rapid and clean coupling between various alpha-aminoalkyl diphenyl phosphonate ester building blocks and commercially available or easily accessible amino acids. These compounds were used for the rapid profiling of dipeptidyl peptidase II (DPP II) and the closely related dipeptidyl peptidase IV (DPP IV). A highly selective DPP II inhibitor was identified, N-cyclopentylglycyl-NHCH(C(6)H(5))PO(OPh)(2) (9.35), that will be useful to discriminate between DPP II and DPP IV in biological systems in order to further elucidate the biological function of DPP II. PMID- 12739953 TI - Structure of a terbium(III)-quinizarine complex: the first crystallographic model for metalloanthracyclines. AB - The crystal structure of a complex of terbium(III) with quinizarine 2-sulfonate has been solved by single-crystal X-ray diffraction methods. The metal cation is coordinated by two adjacent phenolate and quinone oxygens of the anthraquinone moiety of a quinizarine sulfonate anion and by six water molecules. To our knowledge, this is the first structure of a metal complex of the 1,4-dihydroxy anthraquinone ligand. On the basis of strict similarities in the spectroscopic features of the terbium adducts with either doxorubicin or quinizarine 2 sulfonate, the present structure is proposed as a model for the metal complexes of anthracyclines. PMID- 12739954 TI - A new ligand system based on a bipyridine-functionalized calix[4]arene backbone leading to mono- and bimetallic complexes. AB - The synthesis of a new ligand system for mono- and bimetallic complexes based on a calixarene is described. Ligand BBPC (3, bis(bipyridine)-calix[4]arene) is obtained in three steps in 40% overall yield by first brominating one of the methyl groups of the 4,4'-dimethyl-2,2'-bipyridine in two steps and subsequently reacting it with p-tert-butylcalix[4]arene under basic conditions. Reaction of BBPC (3) with 2 equiv of [Rh(NBD)(2)]BF(4) or [Rh(NBD)(CH(3)CN)(2)]BF(4) (NBD = norbornadiene) produces the bimetallic compound BBPC[Rh(NBD)BF(4)](2) (4). Treatment of the ligand with PdCl(2)(CH(3)CN)(2) leads to the isolation of the bimetallic complex BBPC[PdCl(2)](2) (5). When the nickel precursor NiBr(2)(DME) (DME = dimethoxyethane) is reacted with BBPC, the bimetallic complex BBPC[NiBr(2)](2) (6) is isolated which, upon crystallization from methanol, gives the mononuclear bis(bipyridine) complex BBPC[NiBr(OMe)] (7). Full characterization includes X-ray structural studies of complexes 4, 5, and 7. The bimetallic compounds 4 and 5 show metal to metal distances of 4.334 A (for 4) and 3.224 A (for 5). For all three complexes, unique molecular packing arrangements were found, based on hydrophobic/hydrophilic interactions. PMID- 12739955 TI - Novel metal-organic frameworks with specific topology from new tripodal ligands: 1,3,5-tris(1-imidazolyl)benzene and 1,3-bis(1-imidazolyl)-5-(imidazol-1 ylmethyl)benzene. AB - Reactions of two new tripodal ligands 1,3,5-tris(1-imidazolyl)benzene (4) and 1,3 bis(1-imidazolyl)-5-(imidazol-1-ylmethyl)benzene (5) with metal [Ag(I), Cu(II), Zn(II), Ni(II)] salts lead to the formation of novel two-dimensional (2D) metal organic frameworks [Ag(2)(4)(2)][p-C(6)H(4)(COO)(2)].H(2)O (6), [Ag(4)]ClO(4) (7), [Cu(4)(2)(H(2)O)(2)](CH(3)COO)(2).2H(2)O (8), [Zn(4)(2)(H(2)O)(2)](NO(3))(2) (9), [Ni(4)(2)(N(3))(2)].2H(2)O (10), and [Ag(5)]ClO(4) (11). All the structures were established by single-crystal X-ray diffraction analysis. Crystal data for 6: monoclinic, C2/c, a = 23.766(3) A, b = 12.0475(10) A, c = 13.5160(13) A, beta = 117.827(3) degrees, Z = 4. For compound 7: orthorhombic, P2(1)2(1)2(1), a = 7.2495(4) A, b = 12.0763(7) A, c = 19.2196(13) A, Z = 4. For compound 8: monoclinic, P2(1)/n, a = 8.2969(5) A, b = 12.2834(5) A, c = 17.4667(12) A, beta = 96.5740(10) degrees, Z = 2. For compound 9: monoclinic, P2(1)/n, a =10.5699(3) A, b = 11.5037(3) A, c = 13.5194(4) A, beta = 110.2779(10) degrees, Z = 2. For compound 10: monoclinic, P2(1)/n, a = 9.8033(3) A, b = 12.1369(5) A, c = 13.5215(5) A, beta = 107.3280(10) degrees, Z = 2. For compound 11: monoclinic C2/c, a = 18.947(2) A, b = 9.7593(10) A, c = 19.761(2) A, beta = 97.967(2) degrees, Z = 8. Both complexes 6 and 7 are noninterpenetrating frameworks based on the (6, 3) nets, and 8, 9 and 10 are based on the (4, 4) nets while complex 11 has a twofold parallel interpenetrated network with 4.8(2) topology. It is interesting that, in complexes 6,7, and 11 with three-coordinated planar silver(I) atoms, each ligand 4 or 5 connects three metal atoms, while in the case of complexes 8, 9, and 10 with six-coordinated octahedral metal atoms, each ligand 4 only links two metal atoms, and another imidazole nitrogen atom of 4 did not participate in the coordination with the metal atoms in these complexes. The results show that the nature of organic ligand and geometric needs of metal atoms have great influence on the structure of metal-organic frameworks. PMID- 12739956 TI - Ansa versus spiro substitution of cyclophosphazenes: is fluorination essential for ansa to spiro transformation of cyclophosphazenes? AB - Fluorinated ansa substituted cyclophosphazenes endo FcCH(2)P(S)(CH(2)O)(2)[P(F)N](2)(F(2)PN) [Fc = ferrocenyl] (1) and exo FcCH(2)P(S)(CH(2)O)(2)[P(F)N](2)(F(2)PN) (2) readily transform to the spirocyclic compound [FcCH(2)P(S)(CH(2)O)(2)PN](F(2)PN)(2) (3) not only in the presence of CsF but also with non-fluorinated bases such as Cs(2)CO(3), K(2)CO(3), KOBu(t), Et(3)N, DABCO, DBN, and DBU. The analogous tetrachloro ansa compound exo FcCH(2)P(S)(CH(2)O)(2)[P(Cl)N](2)(Cl(2)PN) (5), however, did not transform to the chlorinated spiro compound (6) in the presence of these bases. With excess of CsF, P-Cl bonds of 5 were found to undergo fluorination leading to the formation of 2, which transformed to spirocyclic compound 3. Time dependent (31)P NMR spectroscopy was used to monitor this transformation. Crystal structure studies on the ansa substituted compounds 4 and 5 have shown weak bonding interactions involving C-H...Cl, C-H...O, and C-H...S interactions. PMID- 12739957 TI - NaCl/KCl flux single crystal growth and crystal structure of the new quaternary mixed-metal pnictide: BaCuZn3As3. AB - Synthesis and crystal structure of a new compound, BaCuZn(3)As(3), are reported. Single crystals of BaCuZn(3)As(3) are synthesized via NaCl/KCl flux reaction in a sealed fused silica ampule. Its elemental composition has been determined to be Ba/Cu/Zn/As = 1.03(4):1(0):2.91(6):2.98(3), suggesting BaCuZn(3)As(3) as the chemical formula. The structure of BaCuZn(3)As(3) has been determined by X-ray diffraction. It crystallizes in the orthorhombic Cmcm space group with a = 4.2277(3) A, b = 12.970(1) A, and c = 12.011(1) A at T = 90.7 K, and it exhibits a columnar structure along the a-axis. This structure is isotypic to beta BaCu(4)S(3) but highly distorted. beta-BaCu(4)S(3) is considered to be a layered structure whereas BaCuZn(3)As(3) is a three-dimensional network. PMID- 12739958 TI - Synthesis and properties of rhodium(III) porphyrin cyclic tetramer and cofacial dimer. AB - Rhodium(III) porphyrin complexes, [Rh(4-PyT(3)P)Cl](4) (1) and [Rh(2 PytB(3)P)Cl](2) (2) (4-PyT(3)P = 5-(4-pyridyl)-10,15,20-tritolylporphyrinato dianion, 2-PytB(3)P = 5-(2-pyridyl)-10,15,20-tri(4-tert-butyl)phenylporphyrinato dianion), were self-assembled and characterized by (1)H nuclear magnetic resonance spectroscopy, infrared spectroscopy, and electron spray ionization-mass spectroscopy methods. The spectroscopic results certified that the rhodium porphyrin complexes 1 and 2 have a cyclic tetrameric structure and a cofacial dimeric structure, respectively. The X-ray structure analysis of 1 confirmed the cyclic structure of the complex. The Soret bands of both oligomers were significantly broadened by excitonic interactions between the porphyrin units, compared to those observed for a corresponding analogue of Rh(TTP)(Py)Cl (TTP = 5,10,15,20-tetratolylporphyrinato dianion, Py = pyridine). Stepwise oxidation of the porphyrin rings in the oligomers was observed by cyclic voltammetry. The oligomers 1 and 2 are very stable in solution, and they slowly undergo reactions with pyridine to give corresponding monomer complexes only at high temperatures (approximately 80 degrees C). PMID- 12739959 TI - Synthesis, structure, electrical properties, and band structure calculations of TiAsTe. AB - The new compound TiAsTe has been synthesized by the reaction of the elements in a LiCl/KCl flux at 923 K. The compound crystallizes with four formula units in space group Immm of the orthorhombic system in a cell at 153 K of a = 3.5730(8) A, b = 5.249(1) A, c = 12.794(3) A, V = 240.0(1) A(3). The structure, which is of the NbPS structure type, is a three-dimensional extended framework built from bicapped TiAs(4)Te(4) trigonal prisms. It may be considered to comprise infinity (2) [TiTe] slabs perpendicular to [001] that are interspersed with linear infinity (1)[As] chains running along [010]. The As-As distances alternate at 2.554(2) and 2.695(2) A. Electrical and thermopower measurements indicate that TiAsTe is an n-type metallic compound. Density functional theory calculations help rationalize the chemical bonding and physical properties. PMID- 12739960 TI - Synthesis of mono- and dihalogenated derivatives of (Me2S)2B12H10 and palladium catalyzed boron-carbon cross-coupling reactions of the iodides with grignard reagents. AB - Two series of compounds, 9-X-1,7-(Me(2)S)(2)B(12)H(9) and 9,10-X(2)-1,7 (Me(2)S)(2)B(12)H(8) (X = Cl, Br, I), have been synthesized from reactions of 1,7 (Me(2)S)(2)B(12)H(10) with various halogenating reagents. In addition, reactions of 1,7-(Me(2)S)(2)B(12)H(10) with 2,4-(NO(2))(2)C(6)H(3)SCl and PhSeBr resulted in 9-(2',4'-(NO(2))(2)C(6)H(3)S)-1,7-(Me(2)S)(2)B(12)H(9) and 9,10-(PhSe)(2)-1,7 (Me(2)S)(2)B(12)H(8), respectively. X-ray studies of the dibromo, monoiodo, and aryl thioether derivatives show that electrophilic substitution in 1,7 (Me(2)S)(2)B(12)H(10) takes place at positions 9 and 10, as in the case of the meta-carborane 1,7-C(2)B(10)H(12). From 1,12-(Me(2)S)(2)B(12)H(10) the halides 2 X-1,12-(Me(2)S)(2)B(12)H(9) (X = Br, I) were prepared. For both 1,7- and 1,12 (Me(2)S)(2)B(12)H(10) the best iodination results were obtained using iodine monochloride in refluxing acetonitrile. In the presence of 5 mol % (PPh(3))(2)PdCl(2) the iodides 9-I-1,7-(Me(2)S)(2)B(12)H(9), 2-I-1,12 (Me(2)S)(2)B(12)H(9), and 9,10-I(2)-1,7-(Me(2)S)(2)B(12)H(8) react with RMgX (R = Me, Ph, Bn; X = Cl, Br) in THF to yield the corresponding B-alkyl- and B-aryl substituted products in good yields without using CuI as a cocatalyst. The bromo derivative 9-Br-1,7-(Me(2)S)(2)B(12)H(9) did not react under similar conditions. No interference from the nearby Me(2)S substituent was observed in palladium catalyzed substitution of iodide in 2-I-1,12-(Me(2)S)(2)B(12)H(9). Presumably due to the intramolecular activation of an aryl C-H bond of the benzyl substituent in the intermediate palladium complex, the yield of 9,10-Bn(2)-1,7 (Me(2)S)(2)B(12)H(8) was significantly lower than those of the dimethyl and diphenyl derivatives. The molecular structures of 9-R-1,7-(Me(2)S)(2)B(12)H(9) (R = Ph, Bn) and 2-Bn-1,12-(Me(2)S)(2)B(12)H(9) were obtained by single-crystal X ray analysis. PMID- 12739961 TI - Molecular and electronic structures of iron(II)/(III) complexes containing N,S coordinated, closed-shell o-aminothiophenolato(1-) and o-iminothiophenolato(2-) ligands. AB - The coordination chemistry of the ligands o-aminothiophenol, H(abt), 4,6-di-tert butyl-2-aminothiophenol, H[L(AP)], and 1,2-ethanediamine-N,N'-bis(2 benzenethiol), H(4)('N(2)S(2')), with FeCl(2) under strictly anaerobic and increasingly aerobic conditions has been systematically investigated. Using strictly anaerobic conditions, the neutral, air-sensitive, yellow complexes (mu S,S)[Fe(II)(abt)(2)](2) (1), (mu-S,S)[Fe(II)(L(AP))(2)](2).8CH(3)OH (2), and (mu S,S)[Fe(II)('H(2)N(2)S(2'))](2).CH(3)CN (3) containing high spin ferrous ions have been isolated where (abt)(1-), (L(AP))(1-), and ('H(2)N(2)S(2'))(2-) represent the respective N,S-coordinated, aromatic o-aminothiophenolate derivative of these ligands. When the described reaction was carried out in the presence of trace amounts of O(2) and [PPh(4)]Br, light-green crystals of [PPh(4)][Fe(II)(abt)(2)(itbs)].[PPh(4)]Br (4) were isolated. The anion [Fe(II)(abt)(2)(itbs)](-) contains a high spin ferrous ion, two N,S-coordinated o aminophenolate(1-) ligands, and an S-bound, monoanionic o iminothionebenzosemiquinonate(1-) pi radical, (itbs)(-). Complex 4 possesses an S(t) = 3/2 ground state. In the absence of [PPh(4)]Br and presence of a base NEt(3) and a little O(2), the ferric dimer (mu-NH,NH)[Fe(III)(L(AP))(L(IP))](2) (5a) and its isomer (mu-S,S)[Fe(III)(L(AP))(L(IP))](2) (5b) formed. (L(IP))(2-) represents the aromatic o-iminothiophenolate(2-) dianion of H[L(AP)]. The structures of compounds 2, 4, and 5a have been determined by X-ray crystallography at 100(2) K. Zero-field Mossbauer spectroscopy of 1, 2, 3, and 4 unambiguously shows the presence of high spin ferrous ions: The isomer shift at 80 K is in the narrow range 0.85-0.92 mm s(-1), and a large quadrupole splitting, |DeltaE(Q)|, in the range 3.24-4.10 mm s(-1), is observed. In contrast, 5a and 5b comprise both intermediate spin ferric ions (S(Fe) = 3/2) which couple antiferromagnetically in the dinuclear molecules yielding an S(t) = 0 ground state. PMID- 12739962 TI - Novel chloride-centered discrete CuI8 cubic clusters containing diselenophosphate ligands. Syntheses and structures of [Cu8(mu8-Cl)[Se2P(OR)2](6)](PF6) (R = Et, Pr, iPr)1. AB - Three clusters 1-3, Cu(8)(mu8-Cl)[Se(2)P(OR)(2)](6)(PF(6)) (R= Et, Pr, (i)Pr), were synthesized in high yield from the reaction of [Cu(CH(3)CN)(4)](PF(6)), NH(4)[Se(2)P(OR)(2)], and Bu(4)NCl in a molar ratio of 4:3:1 in diethyl ether. FAB mass spectra show m/z peaks at 2218.10 for 1, 2386.10 for 2, and 2387.34 for 3 which are due to molecular cations, [1-PF(6)]+, [2-PF(6)]+, and [3-PF(6)]+, respectively. (31)P NMR spectra of 1-3 display a singlet at delta 76.48, 76.73, and 69.32 ppm with satellites (J(PSe) = 652, 653, and 648 Hz), respectively. The (77)Se NMR spectra of 1-3 exhibit a doublet peak at -21.7, -16.42, and 2.3 ppm, respectively (J(SeP) = 652 Hz for 1, 653 Hz for 2, and 648 Hz for 3). The X-ray structure (1-3) consists of a discrete cationic cluster in which eight copper ions are linked by six diselenophosphate ligands and a central mu8-Cl ion with a noncoordinating PF(6)(-) anion. The shape of the molecule is a chloride-centered distorted Cu(8) cube in clusters 1 and 2 and a near perfect Cu(8) cube for cluster 3. The dsep ligand exhibits a tetrametallic tetraconnective (mu2, mu2)) coordination pattern, and each occupies a square face of the cube. Each copper atom of the cube is coordinated by three selenium atoms with a strong interaction with the central chloride ion. The observed Cu-Cl distances lie in the range 2.649-2.878 A. PMID- 12739963 TI - Preparation and magnetic properties of Mn(hfac)(2)-complexes of 2-(5-pyrimidinyl) and 2-(3-pyridyl)-substituted nitronyl nitroxides. AB - Mn(hfac)(2) complexes of [2-(5-pyrimidinyl)-4,4,5,5-tetramethyl-4,5-dihydro-1H- imidazoline-1-oxyl 3-oxide] (1) and its 2-(3-pyridyl) analogue (2) were prepared. Both complexes formed similar dimer structures. However, their packing patterns were considerably different. The pyrimidine dimers were aligned to form a linear chain structure, and each dimer was weakly bound by two sets of O6-C2 short contacts. In the pyridine dimer complex, two structurally similar but independent dimers were alternatively arranged, and two dimer-dimer contacts, O6-C2 (3.13 A) and O6-C3 (3.30 A), were observed. The pyrimidine complex showed strong antiferromagnetic behavior in the high temperature region (150-300 K) and weak ferromagnetic behavior below 100 K. Two models were used to analyze these magnetic properties. One is a quintet-septet thermal equilibrium model with mean field approximation, which can reproduce the round minimum observed at about 150 K in chi(p)T plots (J(1)/k(B) = -148 +/- 2 K with theta = +2.5 +/- 0.1 K). The other is a ferromagnetic S = 2 chain model to fit the chi(p)T values in the lower temperature region (J(S=2)/k(B) = +0.31 +/- 0.01 K). The pyridine complex showed antiferromagnetic interactions both in the high and low temperature regions. The magnetic behavior was similarly analyzed with the following parameters: J(1)/k(B) = -140 +/- 2 K with theta = -0.55 +/- 0.05 K, and J(S=2)/k(B) = -0.075 +/- 0.003 K. The ligand-ligand interactions for both of the complexes were theoretically analyzed. The calculated results agreed well with the experiments. The stronger antiferromagnetic behavior observed in both the complexes at high temperatures was attributed to the magnetic interaction between the Mn(II) and the coordinating nitroxide oxygen atom. The weaker ferromagnetic interaction, J(S=2)/k(B) = +0.31 +/- 0.01 K, in the pyrimidine complex was attributed to the coulombic O6-C2 contact. Antiferromagnetic interaction J(S=2)/k(B) = -0.075 +/- 0.003 K in the pyridine complex was attributed to the O6-C3 contact. PMID- 12739964 TI - A rational approach to the design and synthesis of chiral organopalladium-amine complexes. AB - A new chiral auxiliary, (+/-)-N,N-dimethyl-1-(2,5-dimethylphenyl)ethylamine, was designed and synthesized in two steps from 1-acetyl-2,5-dimethylbenzene. Its cyclopalladated dimeric complex could be efficiently resolved via the formation of (S)-prolinate derivatives. Both hand forms of the complex could be obtained in similar yields. Despite the enormous inter-chelate steric constraints, the bulky monodentate ligand 3,4-dimethyl-1-phenylphosphole (DMPP) is able to coordinate regiospecifically to the orthopalladated 2,5-dimethylbenzylamine unit trans to the NMe(2) group. Compared to its naphthylamine analogue, the orthopalladated 2,5 dimethylbenzylamine complex exhibits a significantly higher stereoselectivity in the chiral template promoted asymmetric cycloaddition reaction between DMPP and ethyl vinyl ketone. PMID- 12739965 TI - New structural motifs in the aggregation of neutral gold(I) complexes: structures and luminescence from (alkyl isocyanide)AuCN. AB - The preparation and X-ray crystal structures of (CyNC)Au(I)CN, (n-BuNC)Au(I)CN, and (i-PrNC)Au(I)CN.0.5CH(2)Cl(2) are reported and compared with those of (MeNC)Au(I)CN and (t-BuNC)Au(I)CN, which were previously described. These linear molecules are all organized through aurophilic interactions into three structural classes: simple chains ((CyNC)Au(I)CN and (t-BuNC)Au(I)CN), side-by-side chains in which two strands make Au...Au contact with each other ((n-BuNC)Au(I)CN), and nets in which multiple aurophilic interactions produce layers of gold(I) centers ((i-PrNC)Au(I)CN and (MeNC)Au(I)CN). All of these five solids dissolve to produce colorless, nonluminescent solutions with similar UV/vis spectra. However, each of the solids displays a unique luminescence with emission maxima occurring in the range 371-430 nm. PMID- 12739967 TI - Synthesis and characterization of Nb(OR)4[OC(O)OR] (R = Me, Et, Allyl) and their reaction with the parent alcohol to afford organic carbonates. AB - Nb(OR)(4)[OC(O)OR] (R = Me, Et, allyl) compounds have been synthesized by the reaction of dimeric alkoxo complexes [Nb(OR)(5)](2) with CO(2). The kinetics of the reaction has shown that the monomer in equilibrium with the dimer is the species that reacts with CO(2). The resulting compounds are monomeric with a eta(2)-O,O-bonded hemicarbonate group. The CO(2) uptake is reversible. The hemicarbonates react with the parent alcohol to afford the relevant organic carbonate (RO)(2)CO with a turnover number (TON) of >2. PMID- 12739966 TI - Rh(II) and Rh(I) two-legged piano-stool complexes: structure, reactivity, and electronic properties. AB - The ligand 1,4-bis[4-(diphenylphosphino)butyl]-2,3,5,6-tetramethylbenzene, 3, was used to synthesize a mononuclear Rh(II) complex [(eta(1):eta(6):eta(1)-1,4-bis[4 (diphenylphosphino)butyl]-2,3,5,6-tetramethylbenzene)Rh][PF(6)](2), 6+, in a two legged piano-stool geometry. The structural and electronic properties of this novel complex including a single-crystal EPR analysis are reported. The complex can be cleanly interconverted with its Rh(I) form, allowing for a comparison of the structural properties and reactivity of both oxidation states. The Rh(I) form 6 reacts with CO, tert-butyl isocyanide, and acetonitrile to form a series of 15 membered mononuclear cyclophanes [(eta(1):eta(1)-1,4-bis[4 (diphenylphosphino)butyl]-2,3,5,6-tetramethylbenzene)Rh(CO)(3)][PF(6)] (8), [(eta(1):eta(1)-1,4-bis[4-(diphenylphosphino)butyl]-2,3,5,6 tetramethylbenzene)Rh(CNC(CH(3))(3))(2)][PF(6)] (10), and [(eta(1):eta(1)-1,4 bis[4-(diphenylphosphino)butyl]-2,3,5,6 tetramethylbenzene)Rh(CO)(CH(3)CN)][PF(6)] (11). The Rh(II) complex 6+ reacts with the same small molecules, but over shorter periods of time, to form the same Rh(I) products. In addition, a model two-legged piano-stool complex [(eta(1):eta(6):eta(1)-1,4-bis[3-(diphenylphosphino)propoxy]-2,3,5,6 tetramethylbenzene)Rh][B(C(6)F(5))(4)], 5, has been synthesized and characterized for comparison purposes. The solid-state structures of complexes 5, 6, 6+, and 11 are reported. Structure data for 5: triclinic; P(-)1; a = 10.1587(7) A; b = 11.5228(8) A; c = 17.2381(12) A; alpha = 96.4379(13) degrees; beta = 91.1870(12) degrees; gamma = 106.1470(13) degrees; Z = 2. 6: triclinic; P(-)1; a = 11.1934(5) A; b = 12.4807(6) A; c = 16.1771(7) A; alpha = 81.935(7) degrees; beta = 89.943(1) degrees; gamma = 78.292(1) degrees; Z = 2. 6+: monoclinic; P2(1)/n; a = 11.9371(18) A; b = 32.401(5) A; c = 12.782(2) A; beta = 102.890(3) degrees; Z = 4. 11: triclinic; P(-)1; a = 13.5476(7) A; b = 13.8306(7) A; c = 14.9948(8) A; alpha = 74.551(1) degrees; beta = 73.895(1) degrees; gamma = 66.046(1) degrees; Z = 2. PMID- 12739968 TI - Competition between transferrin and the serum ligands citrate and phosphate for the binding of aluminum. AB - A key issue regarding the speciation of Al(3+) in serum is how well the ligands citric acid and phosphate can compete with the iron transport protein serum transferrin for the aluminum. Previous studies have attempted to measure binding constants for each ligand separately, but experimental problems make it very difficult to obtain stability constants with the accuracy required to make a meaningful comparison between these ligands. In this study, effective binding constants for Al-citrate and Al-phosphate at pH 7.4 have been determined using difference UV spectroscopy to monitor the direct competition between these ligands and transferrin. The analysis of this competition equilibrium also includes the binding of citrate and phosphate as anions to apotransferrin. The effective binding constants are 10(11.59) for the 1:1 Al-citrate complexes and 10(14.90) for the 1:2 Al-citrate complexes. The effective binding constant for the 1:2 Al-phosphate complex is 10(12.02). No 1:1 Al-phosphate complex was detected. Speciation calculations based on these effective binding constants indicate that, at serum concentrations of citrate and phosphate, citrate will be the primary low-molecular-mass ligand for aluminum. Formal stability constants for the Al-citrate system have also been determined by potentiometric methods. This equilibrium system is quite complex, and information from both electrospray mass spectrometry and difference UV experiments has been used to select the best model for fitting the potentiometric data. The mass spectra contain peaks that have been assigned to complexes having aluminum:citrate stoichiometries of 1:1, 1:2, 2:2, 2:3, and 3:3. The difference UV results were used to determine the stability constant for Al(H(-1)cta)-, which was then used in the least-squares fitting of the potentiometric data to determine stability constants for Al(Hcta)+, Al(cta), Al(cta)2(3-), Al(H(-1)cta)(cta)(4-), Al2(H(-1)cta)2(2-), and Al3(H(-1)cta)3(OH)(4-). PMID- 12739970 TI - Influence of hydrogen bonding in competition with lattice interactions on carbonyl coordination at phosphorus. Implications for phosphoryl transfer activated states. AB - A series of phosphorus compounds containing carboxyl groups that serve as mimics for amino acid residues was synthesized. The series was composed of the phosphonium salts 1A, 1B, and 2, the anionic phosphines 3A and 3B, and the anionic phosphine oxide 4. X-ray structural analysis revealed that P-O coordination occurred in the presence of extensive hydrogen bonding and led to pseudo or regular trigonal bipyramidal geometries. (31)P chemical shifts indicated retention of the basic coordination geometries in solution. The two forms observed for 1 and 3 revealed the influence of hydrogen bonding on the P-O donor interactions while 2 and 4 showed the influence of molecular packing effects in competition with hydrogen bonding interactions. The results suggest that phosphoryl transfer enzyme mechanisms should benefit by taking into account P-O donor interactions by residues at active sites that can be manipulated by hydrogen bonding and molecular packing effects in enhancing nucleophilic attack at phosphorus centers. PMID- 12739969 TI - Silylene-bridged dinuclear iron complexes [Cp(OC)2Fe]2SiX2 (X = H, F, Cl, Br, I). Synthesis, molecular structure, vibrational spectroscopy, and theoretical studies. AB - The mu2-silylene-bridged iron complexes [Cp(OC)(2)Fe](2)SiX(2) (X = F (2), Br (4), I (5)) have been prepared from the mu2-SiH(2) functional precursor [Cp(OC)(2)Fe](2)SiH(2) (1) by hydrogen/halogen exchange, using HBF(4), CBr(4), and CH(2)I(2), respectively. The fluoro- and bromo-substituted derivatives 2 and 4 are converted upon UV irradiation to the carbonyl- and dihalosilylene-bridged dinuclear complexes [Cp(OC)Fe](2) (mu2-CO)(mu2-SiX(2)) (X = F (6), Br (7)) via CO elimination. All new compounds have been characterized spectroscopically, and, in addition, the molecular structure of 2, 4, and the previously reported chloro derivative [Cp(OC)(2)Fe](2)SiCl(2) (3) has been determined by single-crystal X ray diffraction methods. For 1-5, the Fourier transform infrared and Raman spectra have been recorded and discussed, together with density functional theory calculations, which support the experimental results of the structural and vibrational analysis. The computed geometries, harmonic vibrational wavenumbers, and their corresponding Raman scattering activities are in good agreement with the experimental data. A significant dependence of the CO and Fe-Si stretching modes on the X substituents of the mu2-silylene bridge has been observed and discussed. PMID- 12739971 TI - Ruthenium(II) and ruthenium(IV) complexes containing kappa1-P-, kappa2-P,O-, and kappa3-P,N,O-iminophosphorane-phosphine ligands Ph2PCH2P[=NP(=O)(OR)2]Ph2 (R = Et, Ph): synthesis, reactivity, theoretical studies, and catalytic activity in transfer hydrogenation of cyclohexanone. AB - [(Ru(eta(6)-p-cymene)(mu-Cl)Cl)(2)] and [(Ru(eta(3):eta(3)-C(10)H(16))(mu Cl)Cl)(2)] react with Ph(2)PCH(2)P[=NP(=O)(OR)(2)]Ph(2) (R = Et (1a), Ph (1b)) affording complexes [Ru(eta(6)-p-cymene)Cl(2)(kappa(1)-P Ph(2)PCH(2)P[=NP(=O)(OR)(2)]Ph(2))] (R = Et (2a), Ph (2b)) and [Ru(eta(3):eta(3) C(10)H(16))Cl(2)(kappa(1)-P-Ph(2)PCH(2)P[=NP(=O)(OR)(2)]Ph(2))] (R = Et (6a), Ph (6b)). While treatment of 2a with 1 equiv of AgSbF(6) yields a mixture of [Ru(eta(6)-p-cymene)Cl(kappa(2)-P,O-Ph(2)PCH(2)P[=NP(=O)(OEt)(2)]Ph(2))][SbF(6)] (3a) and [Ru(eta(6)-p-cymene)Cl(kappa(2)-P,N Ph(2)PCH(2)P[=NP(=O)(OEt)(2)]Ph(2))][SbF(6)] (4a), [Ru(eta(6)-p cymene)Cl(kappa(2)-P,O-Ph(2)PCH(2)P[=NP(=O)(OPh)(2)]Ph(2))][SbF(6)] (3b) and [Ru(eta(3):eta(3)-C(10)H(16))Cl(kappa(2)-P,O Ph(2)PCH(2)P[=NP(=O)(OR)(2)]Ph(2))][SbF(6)] (R = Et (7a), Ph (7b)) are selectively formed from 2b and 6a,b. Complexes [Ru(eta(6)-p-cymene)(kappa(3) P,N,O-Ph(2)PCH(2)P[=NP(=O)(OR)(2)]Ph(2))][SbF(6)](2) (R = Et (5a), Ph (5b)) and [Ru(eta(3):eta(3)-C(10)H(16))(kappa(3)-P,N,O Ph(2)PCH(2)P[=NP(=O)(OR)(2)]Ph(2))][SbF(6)](2) (R = Et (8a), Ph (8b)) have been prepared using 2 equiv of AgSbF(6). The reactivity of 3-5a,b has been explored allowing the synthesis of [Ru(eta(6)-p-cymene)X(2)(kappa(1)-P Ph(2)PCH(2)P[=NP(=O)(OR)(2)]Ph(2))] (R = Et, Ph; X = Br, I, N(3), NCO (9-12a,b)). The catalytic activity of 2-8a,b in transfer hydrogenation of cyclohexanone, as well as theoretical calculations on the models [Ru(eta(6)-C(6)H(6))Cl(kappa(2) P,N-H(2)PCH(2)P[=NP(=O)(OH)(2)]H(2))]+ and [Ru(eta(6)-C(6)H(6))Cl(kappa(2)-P,O H(2)PCH(2)P[=NP(=O)(OH)(2)]H(2))]+, has been also studied. PMID- 12739972 TI - Synthesis of lateral macrobicyclic compartmental ligands: structural, magnetic, electrochemical, and catalytic studies of mono- and binuclear copper(II) complexes. AB - A series of putative mono- and binuclear copper(II) complexes, of general formulas [CuL](ClO(4)) and [Cu(2)L](ClO(4))(2), respectively, have been synthesized from lateral macrocyclic ligands that have different compartments, originated from their corresponding precursor compounds (PC-1, 3,4:9,10-dibenzo 1,12-[N,N'-bis[(3-formyl-2-hydroxy-5-methyl)benzyl]diaza]-5,8 dioxacyclotetradecane; and PC-2, 3,4:9,10-dibenzo-1,12-[N,N'-bis[(3-formyl-2 hydroxy-5-methyl)benzyl]diaza]-5,8-dioxacyclopentadecane). The precursor compound PC-1 crystallized in the triclinic system with space group P(-)1. The mononuclear copper(II) complex [CuL(1a)](ClO(4)) is crystallized in the monoclinic system with space group P2(1)/c. The binuclear copper(II) complex [Cu(2)L(2c)](ClO(4))(2) is crystallized in the triclinic system with space group P(-)1; the two Cu ions have two different geometries. Electrochemical studies evidenced that one quasi-reversible reduction wave (E(pc) = -0.78 to -0.87 V) for mononuclear complexes and two quasi-reversible one-electron-transfer reduction waves (E(1)(pc) = -0.83 to -0.92 V, E(2)(pc) = -1.07 to -1.38 V) for binuclear complexes are obtained in the cathodic region. Room-temperature magnetic-moment studies convey the presence of antiferromagnetic coupling in binuclear complexes [mu(eff) = (1.45-1.55)mu(B)], which is also suggested from the broad ESR spectra with g = 2.10-2.11, whereas mononuclear complexes show hyperfine splitting in ESR spectra and they have magnetic-moment values that are similar to the spin-only value [mu(eff) = (1.69-1.72)mu(B)]. Variable-temperature magnetic susceptibility study of the complex shows that the observed -2J value for the binuclear complex [Cu(2)L(1b)](ClO(4))(2) is 214 cm(-1). The observed initial rate-constant values of catechol oxidation, using complexes as catalysts, range from 4.89 x 10(-3) to 5.32 x 10(-2) min(-1) and the values are found to be higher for binuclear complexes than for the corresponding mononuclear complexes. PMID- 12739973 TI - New rhodium(III) and ruthenium(II) water-soluble complexes with 3,5-diaza-1 methyl-1-azonia-7-phosphatricyclo[3.3.1.1(3,7)]decane. AB - The new water-soluble phosphine complexes of rhodium(III), [RhI(4)(mtpa)(2)]I (1), and ruthenium(II), [RuI(4)(mtpa)(2)].2H(2)O (2) and [RuI(2)(mtpa)(3)(H(2)O)]I(3).2H(2)O (3) (mtpa = 3,5-diaza-1-methyl-1-azonia-7 phosphatricyclo[3.3.1.1(3,7)]decane cation), have been prepared in the reactions of RhCl(3).3H(2)O and RuCl(3).3H(2)O in water in the presence of phosphine and potassium iodide. Properties and reactivity of the complexes have been investigated using (1)H and (31)P NMR and IR spectroscopies. The complexes have also been structurally characterized by single crystal X-ray diffraction studies. The compounds [RhI(4)(mtpa)(2)]I and [RuI(4)(mtpa)(2)].2H(2)O are zwitterionic octahedral complexes. The compounds were tested as catalysts for two-phase hydroformylation of 1-hexene and hydrogenation of cinnamaldehyde. Complex 1 is a selective catalyst for reduction of the C=C bond while complexes 2 and 3 selectively hydrogenate the C=O bond. PMID- 12739974 TI - Synthesis and structural characterization of unprecedented bis-asymmetric heteroscorpionate U(III) complexes: [U(kappa3-H2B(pztBu,Me)(pzMe,tBu))2I] and [U(kappa3-H2B(pztBu,Me)(pzMe2))2I]. AB - [UI(3)(THF)(4)] reacts at room temperature with 2 equiv of KBp(tBu,Me), in toluene, yielding [U(kappa(3)-H(mu-H)B(pz(tBu,Me))(pz(Me,tBu)))(2)I] (1). This unprecedented complex, stabilized by two asymmetric heteroscorpionate ligands, is formed due to an isomerization process promoted in situ by the metal center. To find a general method for preparing this type of compound, we synthesized the novel asymmetric K[H(2)B(pz(tBu,Me))(pz(Me2))], and by a straightforward salt metathesis with [UI(3)(THF)(4)] the novel bis-asymmetric complex [U(kappa(3)-H(mu H)B(pz(tBu,Me))(pz(Me2)))(2)I] (2) was isolated and characterized in the solid state and in solution. As indicated by X-ray crystallographic analysis, the U(III) in 1 and 2 is seven-coordinated by two tridentate asymmetric dihydrobis(pyrazoly)borates and by an iodide. In both cases, the coordination geometry around the metal is very distorted, the pentagonal bipyramid being the one which better describes the arrangement of the atoms around the U(III). An approximate C(2) axis can be defined in the solid state, and is maintained in solution as indicated by the (1)H NMR spectrum of 1 and 2. In the course of attempting to crystallize some of the compounds, monocrystals of the dimer [U(kappa(3)-Bp(tBu,Me))(Hpz(tBu,Me))I(mu-I)](2) (3) were isolated. In this compound each U(III) atom is seven-coordinated by one kappa(3)-Bp(tBu,Me), by one terminal and by two bridging iodide ligands, and by a monodentate Hpz(tBu,Me), exhibiting a distorted 4:3 tetragonal base-trigonal geometry. PMID- 12739975 TI - Preparation and characterization of new ruthenium and osmium containing polyoxometalates, [M(DMSO)3Mo7O24](4-) (M = Ru(II), Os(II)), and their use as catalysts for the aerobic oxidation of alcohols. AB - A new heptamolybdate polyoxometalate structure containing ruthenium(II) or osmium(II) metal centers, [M(II)(DMSO)(3)Mo(7)O(24)](4-) (M = Ru, Os), was synthesized by reaction between (NH(4))(6)Mo(7)O(24) and cis-M(DMSO)(4)Cl(2). X ray structure analysis revealed the complexes to contain a ruthenium/osmium center in a trigonal antiprismatic coordination mode bound to three DMSO moieties via the sulfur atom of DMSO and three oxygen atoms of the new heptamolybdate species. The heptamolybdate consists of seven condensed edge-sharing MoO(6) octahedra with C(2v) symmetry. Three Mo atoms are in classic type II octahedra with a cis dioxo configuration. Two Mo atoms are also type-II-like, but one of the short Mo-O bonds is associated with bridging oxygen atoms rather than terminal oxygen atoms. Two molybdenum atoms are unique in that they are in a trigonally distorted octahedral configuration with three short Mo-O bonds and two intermediate-long M-O bonds and one long Mo-O bond. The [M(II)(DMSO)(3)Mo(7)O(24)](4-) polyoxometalates were effective and in some cases highly selective catalysts for the aerobic oxidation of alcohols to ketones/aldehydes. The integrity of the polyoxometalate was apparently retained at high turnover numbers and throughout the reaction, and a variation of an oxometal type mechanism was proposed to explain the results. PMID- 12739976 TI - Low-valent cobalt complexes with three different pi acceptor ligands: experimental and DFT studies of the reduced and the low-lying excited states of (R-DAB)Co(NO)(CO), R-DAB = substituted 1,4-diaza-1,4-butadiene. AB - The complexes (RN=CH-CH=NR)Co(NO)(CO) with R = isopropyl, 2,6-diisopropylphenyl, or p-tolyl are chemically and electrochemically reducible to radical anions at potentials which strongly depend on R. The DFT calculated structure for the neutral compound with R = iPr agrees with the experiment, and the computed structure of the anion radical reveals changes according to a reduction of the R DAB ligand. EPR results confirm an (R-DAB)-based singly occupied molecular orbital in [(RNCHCHNR)Co(NO)(CO)](.-), with minor but detectable contributions from NO as supported by IR spectroelectrochemistry and as quantified by DFT spin density calculations. The calculations indicate increasingly stabilized CO, NO, and RNCHCHNR pi* acceptor orbitals, in that order. On the basis of TD-DFT (time dependent density functional theory) calculations, the lowest-lying excited states are assigned to metal-to-(R-DAB) charge transfer transitions while bands due to the metal-to-nitrosyl charge transfer occur at higher energies but still in the visible region. Resonance Raman studies were used to probe these assignments. PMID- 12739977 TI - Substituent control of hydrogen bonding in palladium(II)-pyrazole complexes. AB - Inter- and intramolecular hydrogen bonding of an N-H group in pyrazole complexes was studied using ligands with two different groups at pyrazole C-3 and C-5. At C 5, groups such as methyl, i-propyl, phenyl, or tert-butyl were present. At C-3, side chains L-CH(2)- and L-CH(2)CH(2)- (L = thioether or phosphine) ensured formation of chelates to a cis-dichloropalladium(II) fragment through side-chain atom L and the pyrazole nitrogen closest to the side chain. The significance of the ligands is that by placing a ligating side chain on a ring carbon (C-3), rather than on a ring nitrogen, the ring nitrogen not bound to the metal and its attached proton are available for hydrogen bonding. As desired, seven chelate complexes examined by X-ray diffraction all showed intramolecular hydrogen bonding between the pyrazole N-H and a chloride ligand in the cis position. In addition, however, intermolecular hydrogen bonding could be controlled by the substituent at C-5: complexes with either a methyl at C-5 or no substituent there showed significant intermolecular hydrogen bonding interactions, which were completely avoided by placing a tert-butyl group at C-5. The acidity of two complexes in acetonitrile solutions was estimated to be closer to that of pyridinium ion than those of imidazolium or triethylammonium ions. PMID- 12739978 TI - New disulfido molybdenum-manganese complexes exhibit facile addition of small molecules to the sulfur atoms. AB - The reaction of Mn(2)(CO)(7)(mu-S2) (1) with [CpMo(CO)(3)](2) (Cp = C(5)H(5)) and [Cp*Mo(CO)(3)](2) (Cp* = C(5)(CH(3))(5)) yielded the new mixed-metal disulfide complexes CpMoMn(CO)(5)(mu-S2) (2) and Cp*MoMn(CO)(5)(mu-S2) (3) by a metal-metal exchange reaction. Compounds 2 and 3 both contain a bridging disulfido ligand lying perpendicular to the Mo-Mn bond. The bond distances are Mo-Mn = 2.8421(10) and 2.8914(5) A and S-S = 2.042(2) and 1.9973(10) A for 2 and 3, respectively. A tetranuclear metal side product CpMoMn(3)(CO)(13)(mu3-S)(mu4-S) (4) was also isolated from the reaction of 1 with [CpMo(CO)(3)](2). Compounds 2 and 3 react with CO to yield the dithiocarbonato complexes CpMoMn(CO)(5)[mu-SC(=O)S] (5) and Cp*MoMn(CO)(5)[mu-SC(=O)S] (6) by insertion of CO into the S-S bond. Similarly, tert-butylisocyanide was inserted into the S-S bond of 2 and 3 to yield the complexes CpMoMn(CO)(5)[mu-S(C=NBu(t))S] (7) and Cp*MoMn(CO)(5)[mu-S(C=NBu(t))S] (8), respectively. Ethylene and dimethylacetylene dicarboxylate also inserted into the S-S bond of 2 and 3 at room temperature to yield the ethanedithiolato ligand bridged complexes CpMoMn(CO)(5)(mu-SCH(2)CH(2)S) (9), Cp*MoMn(CO)(5)(mu SCH(2)CH(2)S) (10), CpMoMn(CO)(5)[mu-SC(CO(2)Me)=C(CO(2)Me)S] (11), and Cp*MoMn(CO)(5)[mu-SC(CO(2)Me)=C(CO(2)Me)S] (12). Allene was found to insert into the S-S bond of 2 by using one of its two double bonds to yield the complex CpMoMn(CO)(5)[mu-SCH(2)C(=CH(2))S] (13). The molecular structures of the new complexes 2-7 and 9-13 were established by single-crystal X-ray diffraction analyses. PMID- 12739979 TI - Oxamato-bridged trinuclear NiIICuIINiII complexes: a new (NiIICuIINiII)2 hexanuclear complex and supramolecular structures. Characterization and magnetic properties. AB - Eight oxamato-bridged heterotrinuclear Ni(II)Cu(II)Ni(II) complexes of formula ([Ni(H(2)O)(dpt)](2)(mu-Cu(H(2)O)(opba)))(ClO(4))2 (1), ([Ni(H(2)O)(dien)](2)(mu Cu(pba)))(ClO(4))(2).6H(2)O (2), ([Ni(H(2)O)(Medpt)](2)(mu Cu(OHpba)))(ClO(4))(2).4H(2)O (3), ([Ni(H(2)O)(dien)](2)(mu Cu(Me(2)pba)))(ClO(4))(2).2.5H(2)O (4), ([Ni(H(2)O)(dpt)](2)(mu Cu(Me(2)pba)))(ClO(4))(2).2H(2)O (5), ([Ni(H(2)O)(dien)](2)(mu Cu(OHpba)))(ClO(4))(2).4H(2)O (6), ([Ni(2)(dpt)(2)(mu-Cu(H(2)O)(pba))](2)(mu N(3))(2))Na(2)(ClO(4))(4).6H(2)O (7), and ([Cu(H(2)O)(2)(dpt)Ni(2)(H(2)O)(dpt)(2)](mu-H(2)Me(2)pba(2-)))(ClO(4))(4).3H(2)O (8) in which opba = o-phenylenbis(oxamato), pba = 1,3-propylenebis(oxamato), OHpba = 2-hydroxy-1,3-propylenebis(oxamato), Me(2)pba = 2,2-dimethyl-1,3 propylenbis(oxamato), dpt = 3,3'-diaminodipropylamine, dien = 2,2' diaminodiethylamine, and Medpt = 3,3'-diamino-N-methyldipropylamine were synthesized and characterized. The crystal structures of 1, 7, and 8 were solved. For complex 1, the trinuclear entities are linked by hydrogen bonds forming a one dimensional system, and for complex 8, the presence of van der Waals interactions gives a one-dimensional system, too. For complex 7, the trinuclear entities are self-assembled by azido ligands, given a hexanuclear system; each of these hexanuclear entities are self-assembled through two [Na(O)(3)(H(2)O)(3)] octahedral-sharing one-edge entities, given a one-dimensional system. The magnetic behavior of complexes 2-7 was investigated by variable-temperature magnetic susceptibility measurements. Complexes 2-6 exhibit the minimum characteristic of this kind of polymetallic species with an irregular spin state structure. The Jvalue through the oxamato bridge varied between -88 cm(-1) (for 6) and -111.2 cm(-1) (for 5). For complex 7, the values obtained were J(1) = 101.7 cm(-1) (through the oxamato ligand) and J(2) = -3.2 cm(-1) (through the azido ligand). PMID- 12739980 TI - Iron-promoted nucleophilic additions to diimine-type ligands: a synthetic and structural study. AB - We report here three examples of the reactivity of protic nucleophiles with diimine-type ligands in the presence of Fe(II) salts. In the first case, the iron promoted alcoholysis reaction of one nitrile group of the ligand 2,3-dicyano-5,6 bis(2-pyridyl)-pyrazine (L1) permitted the isolation of an stable E-imido-ester, [Fe(L1')(2)](CF(3)SO(3))(2) (1), which has been characterized by spectroscopic studies (IR, ES-MS, Mossbauer), elemental analysis, and crystallographically. Compound 1 consists of mononuclear octahedrally coordinated Fe(II) complexes where the Fe(II) ion is in its low-spin state. The iron-mediated nucleophilic attack of water to the asymmetric ligand 2,3-bis(2-pyridyl)pyrido[3,4-b]pyrazine (L2) has also been studied. In this context, the crystal structures of two hydration-oxidation Fe(III) products, [Fe(L2')(2)](ClO(4))(3).3CH(3)CN (2) and trans-[FeL2"Cl(2)] (3), are described. Compounds 2 and 3 are both mononuclear Fe(III) complexes where the metals occupy octahedral positions. In principle, L2 is expected to coordinate to metal ions through its bipyridine-type units to form a five-membered ring; however, this is not the case in compounds 2 and 3. In 2, the ligand coordinates through its pyridines and through the hydroxyl group attached to the pyrazine imino carbon after hydration, that is, in an N,O,N tridentate manner. In compound 3, the ligand has suffered further transformations leading to a very stable diamido complex. In this case, the metal ion achieves its octahedral geometry by means of two pyridines, two amido N atoms, and two axial chlorine atoms. Magnetic susceptibility measurements confirmed the spin state of these two Fe(III) species: compounds 2 and 3 are low-spin and high-spin, respectively. PMID- 12739981 TI - Pharmacokinetic changes during extracorporeal membrane oxygenation: implications for drug therapy of neonates. AB - Extracorporeal membrane oxygenation (ECMO) is a prolonged form of cardiopulmonary bypass used to support patients with life-threatening respiratory or cardiac failure. In neonates, ECMO is used for a variety of indications, including sepsis and pulmonary diseases such as meconium aspiration syndrome, persistent pulmonary hypertension or congenital diaphragmatic hernia. In recent years, ECMO has been increasingly used after surgery to correct congenital cardiac defects. Despite the need for numerous drugs to maintain the ECMO circuit and treat the patient's underlying illness, relatively little is known of the disposition of drugs in this patient population. To date, the largest number of pharmacokinetic studies have been conducted with gentamicin and vancomycin. Both drugs have been found to have an increased volume of distribution, probably as a result of the addition of a large exogenous blood volume for circuit priming. Elimination half-lives for both drugs are prolonged during ECMO, with several studies demonstrating a return to expected values after decannulation. The reason for this prolonged elimination is probably multifactorial, with a reduction in renal function as the primary determinant. This same pattern of an increased volume of distribution and prolonged elimination has been found for several other drugs, including tobramycin, bumetanide and ranitidine. Other factors that affect drug disposition during ECMO include loss of the drug from adhesion to the circuit components and loss in the circulating blood volume during changes in the equipment. The benzodiazepines and propofol are largely sequestered within the circuit. Serum concentrations of heparin, morphine, fentanyl, furosemide, phenytoin and phenobarbital are also reduced by these mechanisms. The addition of haemofiltration or dialysis in up to a quarter of ECMO patients further complicates the determination of population pharmacokinetic parameters. The literature published to date on the pharmacokinetic changes associated with ECMO provide preliminary support for dosage adjustment; however, more research is needed to identify optimal administration strategies for this patient population. PMID- 12739982 TI - Pharmacokinetics of pegylated liposomal Doxorubicin: review of animal and human studies. AB - Pegylated liposomal doxorubicin (doxorubicin HCl liposome injection; Doxil or Caelyx) is a liposomal formulation of doxorubicin, reducing uptake by the reticulo-endothelial system due to the attachment of polyethylene glycol polymers to a lipid anchor and stably retaining drug as a result of liposomal entrapment via an ammonium sulfate chemical gradient. These features result in a pharmacokinetic profile characterised by an extended circulation time and a reduced volume of distribution, thereby promoting tumour uptake. Preclinical studies demonstrated one- or two-phase plasma concentration-time profiles. Most of the drug is cleared with an elimination half-life of 20-30 hours. The volume of distribution is close to the blood volume, and the area under the concentration-time curve (AUC) is increased at least 60-fold compared with free doxorubicin. Studies of tissue distribution indicated preferential accumulation into various implanted tumours and human tumour xenografts, with an enhancement of drug concentrations in the tumour when compared with free drug. Clinical studies of pegylated liposomal doxorubicin in humans have included patients with AIDS-related Kaposi's sarcoma (ARKS) and with a variety of solid tumours, including ovarian, breast and prostate carcinomas. The pharmacokinetic profile in humans at doses between 10 and 80 mg/m(2) is similar to that in animals, with one or two distribution phases: an initial phase with a half-life of 1-3 hours and a second phase with a half-life of 30-90 hours. The AUC after a dose of 50 mg/m(2) is approximately 300-fold greater than that with free drug. Clearance and volume of distribution are drastically reduced (at least 250-fold and 60-fold, respectively). Preliminary observations indicate that utilising the distinct pharmacokinetic parameters of pegylated liposomal doxorubicin in dose scheduling is an attractive possibility. In agreement with the preclinical findings, the ability of pegylated liposomes to extravasate through the leaky vasculature of tumours, as well as their extended circulation time, results in enhanced delivery of liposomal drug and/or radiotracers to the tumour site in cancer patients. There is evidence of selective tumour uptake in malignant effusions, ARKS skin lesions and a variety of solid tumours. The toxicity profile of pegylated liposomal doxorubicin is characterised by dose-limiting mucosal and cutaneous toxicities, mild myelosuppression, decreased cardiotoxicity compared with free doxorubicin and minimal alopecia. The mucocutaneous toxicities are dose-limiting per injection; however, the reduced cardiotoxicity allows a larger cumulative dose than that acceptable for free doxorubicin. Thus, pegylated liposomal doxorubicin represents a new class of chemotherapy delivery system that may significantly improve the therapeutic index of doxorubicin. PMID- 12739984 TI - Transdermal drug delivery of insulin with ultradeformable carriers. AB - For a long time, scientists believed that macromolecules can only be introduced through the skin with a hypodermic needle or some other harsh treatment that locally damages the skin barrier. It is now clear that macromolecules can be administered epicutaneously, so that insulin, for example, can exhibit therapeutic effects in patients with type 1 diabetes mellitus. When carriers are employed for the purpose, the drugs must be associated with specifically designed vehicles in the form of highly deformable aggregates and applied on the skin non occlusively. Using such optimised carriers, so-called Transfersomes, ensures reproducible and efficient transcutaneous carrier and drug transport. Insulin loaded Transfersomes, for example, can deliver the drug through the non compromised skin barrier with a reproducible drug effect that resembles closely that of an ultralente insulin injected under the skin; the pharmacokinetic and pharmacodynamic properties of the injected and transdermal insulin are also comparable. The efficacy of transcutaneously delivered insulin in Transfersomes is not affected by the previous therapy, similar results having been measured in patients normally receiving intensified insulin therapy or a continuous subcutaneous infusion of insulin solution. Systemic normoglycaemia that lasts at least 16 hours has been achieved using a single non-invasive, epicutaneous administration of insulin in Transfersomes. Experience with other drugs suggests that the biodistribution of injected and transcutaneously delivered drugs can be very similar. This notwithstanding, Transfersomes can be designed and applied so as to mediate site-specific drug delivery into peripheral musculoskeletal tissues or into the skin, as may be desired. PMID- 12739985 TI - No influence of ethnic origin on the pharmacokinetics and pharmacodynamics of melagatran following oral administration of ximelagatran, a novel oral direct thrombin inhibitor, to healthy male volunteers. AB - OBJECTIVE: To determine the influence of ethnic origin on the pharmacokinetic and pharmacodynamic properties of melagatran after oral administration of ximelagatran, a novel oral direct thrombin inhibitor. STUDY DESIGN: This was an open-label, non-randomised study with a single study session. SUBJECTS: Thirty six young healthy male subjects living in France were divided equally according to their ethnic origin (African, Asian and Caucasian). METHODS: All subjects received a single 50mg oral dose of ximelagatran in solution. Blood and urine samples for pharmacokinetic evaluation were collected up to 12 and 24 hours after administration, respectively. Blood samples were also collected to determine the activated partial thromboplastin time (APTT), an ex vivo coagulation time measurement used to demonstrate inhibition of thrombin, up to 24 hours after administration. RESULTS: The absorption of ximelagatran, and its bioconversion to melagatran, was rapid in all three ethnic groups. The metabolite pattern in plasma and urine was similar in all groups, with melagatran being the dominant compound. For ximelagatran, the mean area under the plasma concentration-time curve (AUC) was similar in the three groups, suggesting that there was no difference in the extent to which ximelagatran was absorbed. Melagatran AUC was higher in the Asian subjects, with a mean Asian/Caucasian ratio (95% CI) of 1.23 (1.04, 1.45). This was presumably because of their lower bodyweight, which is correlated to lower renal function. Following normalisation for bodyweight, there were no statistically significant differences between the three ethnic groups. This finding suggests that renal elimination was lower for Asian subjects, whereas there were no differences in the conversion of ximelagatran to melagatran. The interindividual variability of melagatran AUC was low (coefficient of variation 19-26%), and the mean bioavailability of melagatran, estimated using a mean value for melagatran clearance obtained from Caucasian subjects in a previous study, was approximately 20% in all groups (range of mean values 19-23%). APTT increased nonlinearly with increasing melagatran plasma concentration, and no difference in the concentration-response relationship was observed between the groups. CONCLUSIONS: After oral administration of ximelagatran, the pharmacokinetic and pharmacodynamic properties of melagatran are independent of ethnic origin. The elimination of melagatran is correlated with renal function. PMID- 12739986 TI - No influence of obesity on the pharmacokinetics and pharmacodynamics of melagatran, the active form of the oral direct thrombin inhibitor ximelagatran. AB - BACKGROUND: Ximelagatran, an oral direct thrombin inhibitor, is currently in clinical development for the prevention and treatment of thromboembolic disease. Following oral administration, ximelagatran undergoes rapid bioconversion to its active form, melagatran, via two minor intermediates. Obesity, defined as body mass index (BMI) >30 kg/m(2), is a recognised risk factor for thrombosis. There is potential for differences in the pharmacokinetics and pharmacodynamics of drugs administered to obese versus non-obese patients, and some drugs may require alternative administration strategies in obese patients. OBJECTIVE: To investigate the effect of obesity on the pharmacokinetics and pharmacodynamics of melagatran after oral administration of ximelagatran. DESIGN AND PARTICIPANTS: This was an open-label, single-dose, group-matched study in which obese subjects (BMI 32-39 kg/m(2); six male and six female; age 21-40 years) were matched by sex and age (+/-2 years) with non-obese subjects (BMI 21-26 kg/m(2); six male and six female; aged 21-39 years). Each subject received a single oral dose of ximelagatran 24mg. Blood samples for determination of plasma concentrations of melagatran and activated partial thromboplastin times (APTT; a marker of melagatran pharmacodynamics) were collected up to 12 hours after administration. RESULTS: There were no statistically significant differences in the pharmacokinetic properties of melagatran between obese and non-obese subjects. Values of area under the melagatran plasma concentration-time curve, maximum plasma concentration (C(max)), time at which C(max) occurred and terminal elimination half-life were approximately 1 micromol. h/L, 0.2 micromol/L, 2 hours and 3 hours in both obese and non-obese subjects, respectively. In addition, there was no statistically significant difference between the obese and non-obese subjects in the amount of ximelagatran, melagatran or the minor intermediates ethyl-melagatran and melagatran hydroxyamidine excreted in urine. When relating the prolongation of APTT ratio to the square root of plasma concentration of melagatran and obesity status (no/yes), no statistically significant interaction between plasma concentration and obesity status was observed. Ximelagatran was well tolerated in both obese and non-obese subjects, and no bleeding events or serious adverse events occurred. CONCLUSIONS: No differences in the pharmacokinetics or pharmacodynamics of melagatran were detected between obese and non-obese subjects after oral administration of ximelagatran, suggesting that dose adjustment of ximelagatran in obesity (BMI up to 39 kg/m(2)) is not necessary. PMID- 12739987 TI - Aminoglycoside dosages and nephrotoxicity: quantitative relationships. AB - OBJECTIVE: To develop a model that relates the probability of occurrence of nephrotoxicity to the cumulative area under the curve (AUC) of amikacin serum concentration. DESIGN AND PATIENTS: This was a retrospective study of two groups of patients in whom nephrotoxicity was observed after administration of amikacin. The first group consisted of patients treated with once-daily administration (ODA) [n = 13]. The second group consisted of patients treated with twice-daily administration (TDA) [n = 22]. MAIN OUTCOME MEASURES: The probability of nephrotoxicity occurrence. RESULTS: The model is a powerful tool to represent and describe the influence of the dosage regimen on aminoglycoside nephrotoxicity. The onset of nephrotoxicity is delayed in the ODA group (p = 0.01) for the same total daily dose among the two groups. The cumulative serum AUC values at onset of nephrotoxicity were greater for the ODA group (p = 0.029). In addition, for the same probability of nephrotoxicity occurrence (50%), the cumulative AUC for the ODA dosage regimen is 2 613 mg. h/L versus only 1 521 mg. h/L for the TDA dosage regimen. The difference in nephrotoxicity between ODA and TDA is greatest for a cumulative AUC of 2 495 mg. h/L, which corresponds to standard therapy with amikacin 900 mg/day during a 7-day period, i.e. 15 mg/kg/day for a 60kg patient with normal renal function (initial creatinine clearance >80 mL/min). For an AUC above 2 495 mg. h/L, the difference in nephrotoxicity decreases slowly to zero. This result means that ODA is especially justified when the treatment is administered over a short duration, i.e. less than 7 days. CONCLUSIONS: The utility of selecting ODA in order to obtain less nephrotoxicity in comparison with TDA is therefore not established when the treatment is prolonged. In clinical use, the choice of the dosage regimen is not clear-cut, and both expected efficacy and expected toxicity must be taken into account in order to obtain an overall optimisation of each patient's therapy. PMID- 12739988 TI - Current best treatment for non-alcoholic fatty liver disease. AB - Treatment of patients with non-alcoholic fatty liver disease (NAFLD) has typically been focused on the management of associated conditions such as obesity, diabetes mellitus and hyperlipidaemia. NAFLD associated with obesity may be resolved by weight reduction, although the benefits of weight loss have been inconsistent. Improving insulin sensitivity with lifestyle modifications or medications usually improves glucose and lipid levels in patients with diabetes and hyperlipidaemia. Improving insulin sensitivity is expected to improve the liver disease but in many diabetic/hyperlipidaemic patients with NAFLD, the appropriate control of glucose and lipid levels is not always accompanied by improvement of the liver condition. Results of pilot studies evaluating ursodeoxycholic acid, gemfibrozil, betaine, N-acetylcysteine, alphatocopherol, metformin and thiazolidinedione derivatives suggest that these medications may be of potential benefit. This article reviews the treatment modalities currently available for patients with NAFLD, including emerging data from clinical trials evaluating promising medications as well as possibilities for the future. PMID- 12739983 TI - Clinical pharmacokinetics of antioxidants and their impact on systemic oxidative stress. AB - Dietary antioxidants play a major role in maintaining the homeostasis of the oxidative balance. They are believed to protect humans from disease and aging. Vitamin C (ascorbic acid), vitamin E (tocopherol), beta-carotene and other micronutrients such as carotenoids, polyphenols and selenium have been evaluated as antioxidant constituents in the human diet. This article addresses data provided from clinical trials, highlighting the clinical pharmacokinetics of vitamin C, vitamin E, beta-carotene, lycopene, lutein, quercetin, rutin, catechins and selenium. The bioavailability of vitamin C is dose-dependent. Saturation of transport occurs with dosages of 200-400 mg/day. Vitamin C is not protein-bound and is eliminated with an elimination half-life (t((1/2))) of 10 hours. In Western populations plasma vitamin C concentrations range from 54-91 micro mol/L. Serum alpha- and gamma-tocopherol range from 21 micro mol/L (North America) to 27 micro mol/L (Europe) and from 3.1 micro mol/L to 1.5 micro mol/L, respectively. alpha-Tocopherol is the most abundant tocopherol in human tissue. The bioavailability of all-rac-alpha-tocopherol is estimated to be 50% of R,R,R alpha-tocopherol. The hepatic alpha-tocopherol transfer protein (alpha-TTP) together with the tocopherol-associated proteins (TAP) are responsbile for the endogenous accumulation of natural alpha-tocopherol. Elimination of alpha tocopherol takes several days with a t((1/2)) of 81 and 73 hours for R,R,R-alpha tocopherol and all-rac-alpha-tocopherol, respectively. The t((1/2)) of tocotrienols is short, ranging from 3.8-4.4 hours for gamma- and alpha tocotrienol, respectively. gamma-Tocopherol is degraded to 2, 7, 8-trimethyl-2 (beta-carboxyl)-6-hyrdoxychroman by the liver prior to renal elimination. Blood serum carotenoids in Western populations range from 0.28-0.52 micro mol/L for beta-carotene, from 0.2-0.28 for lutein, and from 0.29-0.60 for lycopene. All trans-carotenoids have a better bioavailability than the 9-cis-forms. Elimination of carotenoids takes several days with a t((1/2)) of 5-7 and 2-3 days for beta carotene and lycopene, respectively. The bioconversion of beta-carotene to retinal is dose-dependent, and ranges between 27% and 2% for a 6 and 126mg dose, respectively. Several oxidised metabolites of carotenoids are known. Flavonols such as quercetin glycosides and rutin are predominantly absorbed as aglycones, bound to plasma proteins and subsequently conjugated to glucuronide, sulfate, and methyl moieties. The t((1/2)) ranges from 12-19 hours. The bioavailabillity of catechins is low and they are eliminated with a t((1/2)) of 2-4 hours. Catechins are degraded to several gamma-valerolactone derivatives and phase II conjugates have also been identified. Only limited clinical pharmacokinetic data for other polyphenols such as resveratrol have been reported to date. PMID- 12739989 TI - Current pharmacotherapy in the management of cirrhosis: focus on the hyperdynamic circulation. AB - Many major complications of hepatic cirrhosis relate to the development of a characteristic hyperdynamic circulatory state in these patients, irrespective of the underlying disease aetiology. Vasodilatation of the systemic and splanchnic circulations leads to a reduced total systemic vascular resistance, increased cardiac output and intense activation of neurohumoral vasoconstrictor systems including the sympathetic nervous system, renin-angiotensin system and vasopressin. Vasoconstriction of the renal and hepatic circulations contributes to the development of renal failure and portal hypertension, respectively. Current treatments that focus on amelioration of these circulatory derangements offer much promise, however, they are often limited by side effects in these patients. PMID- 12739990 TI - Radiocontrast-induced nephropathy and percutaneous coronary intervention: a review of preventive measures. AB - Injectable and absorbable contrast media for the use in radiology, all of which contains iodine as an essential component, has been, and continues to be, one of the main sources of agents which cause hospital-acquired renal failure. Although numerous methods have been explored to prevent renal contrast damage, radiocontrast-induced nephropathy continues to be a concern in patients with existing renal insufficiency, who undergo contrast-enhanced radiographic examinations. Patients who develop contrast-induced nephropathy (CIN) have a worse prognosis and an increased risk of complications and mortality. Prevention of CIN during radiocontrast procedures continues to elude clinicians and is a chief concern during percutaneous coronary intervention, as these patients often have multiple comorbidities. A wide variety of animal and clinical investigations, and substances have been tried in order to prevent this complication, including: dialysis, contrast volume and type; adenosine antagonists; acetylcysteine; fenoldopam; and various others. The purpose of this review is to appraise all the past and current strategies employed to prevent CIN, especially during percutaneous coronary intervention. PMID- 12739992 TI - Effect of hormone replacement therapy on cardiovascular disease: current opinion. AB - Cardiovascular disease (CVD) is increasingly being recognised as having a profound effect on women, especially after menopause. Lack of oestrogen has been targeted as one of the reasons for increased incidence of CVD in postmenopausal women. Oestrogen has been found to have favourable effects on lipid profile, tone of vascular smooth muscle cells and fibrinogen levels. Several observational studies have supported these experimental findings, consistently demonstrating reduced cardiovascular risks in users of hormone replacement therapy (HRT). However, evidence from recent clinical trials has challenged this widespread belief. Heart and Estrogen/Progesterone Replacement Study II, Estrogen Replacement and Atherosclerosis trial and more recently, Women's Health Initiative, have shown that HRT has no role in primary and secondary prevention of CVD and most authorities currently do not advocate HRT for the prevention of CVD. PMID- 12739991 TI - Direct thrombin inhibitors. AB - This review deals with a newly-developed category of antithrombotic drugs - the direct thrombin inhibitors. These agents interact with thrombin and block its catalytic activity on fibrinogen, platelets and other substrates. Heparin and its derivatives (low molecular weight heparins and the active pentasaccharide) inhibit thrombin and/or other coagulation serine proteases indirectly via antithrombin, and the warfarin-type drugs interfere with the synthesis of the precursors of the coagulation serine proteases. The direct thrombin inhibitors approved for clinical use at present (lepirudin, desirudin, bivalirudin, argatroban) and another in the advanced clinical testing stage (melagatran/ximelagatran), are the subject of this review. The chemical structure; kinetics of thrombin inhibition; pharmacokinetics and clinical use of each of these is discussed. PMID- 12739993 TI - Sex differences in heart disease incidence and prevalence: implications for intervention. AB - Women present clinically with coronary disease later in life than men do, and given their increased life expectancy compared to men, represent an increasingly larger proportion of patients with coronary disease. Coronary disease in women results in a large human and financial burden to the healthcare system. Effective prevention measures should integrate the clinical and social features of coronary disease which are specific to women and should be anchored within a comprehensive understanding of the burden of coronary disease in women. While attention has been recently directed at coronary disease in women, there is relatively limited information on the incidence and prevalence of coronary disease in women and how it may have changed over time; few studies include sufficient numbers of women to draw appropriate inference. This paper reviews the epidemiology of coronary disease in women and discusses the implications for intervention. PMID- 12739994 TI - Pegylated interferon with ribavirin therapy for chronic infection with the hepatitis C virus. AB - Chronic infection with the hepatitis C virus is common. In the past, therapy involved a combination of thrice-weekly interferon (IFN) injections combined with oral ribavirin. This therapy was expensive, poorly tolerated and poorly effective, only curing approximately 40% of treated patients. Long-acting IFNs have recently been developed by linking IFN to polyethylene glycol and these 'pegylated' IFNs are now the standard of care for patients with chronic hepatitis C (CHC). Two pegylated (PEG) IFNs are available; 40 kDa PEG-IFNalpha(2a) (Pegasys, Hoffmann-La Roche) and the 12 kDa PEG-IFN-alpha(2b) (Peg-Intron, Schering-Plough). They have different physicochemical, pharmacokinetic and pharmacodynamic properties. The 40 kDa PEG-IFN-alpha(2a) is dispensed as a solution and used at a fixed dose whereas the 12 kDa PEG-IFN-alpha(2b) is a dry powder, which is reconstituted prior to administration, and the dose is dependent upon body weight. Both PEG-IFNs are given by a once-weekly injection and as monotherapy, they are more effective than standard IFN-alpha. The 40 kDa PEGIFN alpha(2a) cures 36 - 39% of patients and the 12 kDa pegylated-IFN-alpha(2b) cures 23 - 25%. When combined with ribavirin, the two PEG-IFNs have acceptable safety profiles and cure > 50% of treated patients (56 and 54% for the 40 kDa PEG-IFN alpha(2a) and 12 kDa PEG-IFN-alpha(2b), respectively). For the 40 kDa PEG-IFN alpha(2a) it is possible to predict the outcome of therapy after 12 weeks of treatment. The new PEG-IFNs are a significant advance in the therapy of CHC infection. Their ease of administration, coupled with their improved efficacy, is likely to lead to an increase in the proportion of infected patients who wish to receive treatment. PMID- 12739995 TI - Best pharmacological practice: urinary tract infections. AB - Urinary tract infection is the most frequent bacterial infection. Acute uncomplicated urinary infection and acute non-obstructive pyelonephritis occur in young women with normal genitourinary tracts. Empirical short-course therapy is preferred for the management of acute cystitis, but evolving resistance requires continuing reassessment of optimal antimicrobial selection. Empirical trimethoprim or trimethoprim/sulfamethoxazole has been recommended, but increasing resistance to these agents suggests that pivmecillinam, nitrofurantoin and perhaps fosfomycin trometamol should be considered. Although flouroquinolones are effective as short-course therapy, widespread empirical use of these agents should be discouraged because of potential promotion of resistance. For acute non obstructive pyelonephritis, flouroquinolones are the empirical oral treatment of choice, although urine culture results should direct continuing therapy. Complicated urinary tract infection occurs in men or women of all ages with underlying abnormalities of the genitourinary tract. Treatment of complicated urinary infection is individualised, taking into consideration the underlying abnormality and susceptibilities of the infecting organism. Asymptomatic bacteriuria should not be treated except in pregnant women, in patients prior to undergoing an invasive surgical procedure, or renal transplant recipients in the early postrenal transplant period. PMID- 12739997 TI - Pharmacotherapy of first-episode psychosis. AB - Early intervention in psychosis has attracted more attention in the last few years. The treatment of this phase of the disorders requires a specific and adapted approach. The issue of engaging the patient is so critical that it influences not only the choice of medication, but also the context and the way in which it is administered. In the case of a first admission, patients should be observed for 24-48 h without any antipsychotic treatment, in order to clarify the diagnosis and exclude the possibility that symptoms are caused by acute intoxication with illicit substances, for example. The diagnosis is often difficult and unstable. A dimensional, rather than a categorical approach, is usually more likely to be adopted. In recent years, atypical antipsychotics have become the most frequently used first-line treatment. They are less likely to cause secondary negative symptoms, cognitive impairments and dysphoria. They also appear to influence the course of depression and hostility/aggression better than conventional neuroleptics, have possibly mood-stabilising properties and, subjectively, are often better accepted by patients. On the risk side, prevalence of acute extrapyramidal side effects and possibly tardive dyskinesia are lower, compared to the older neuroleptics. Although, the risk for short-term weight gain, cardiovascular, and especially hyperglycaemic complications are somewhat higher for some of these antipsychotics. Finally, the dose should be adapted as it has been shown that patients presenting a first psychotic episode respond to a lower dose of antipsychotic. This article focuses on the pharmacotherapy of first episode psychosis, on the basis of a computerised and a manual search for articles dealing with antipsychotic treatment of these patients. Findings are discussed and combined in clinical guidelines for first-episode affective and non affective psychosis, for patients with incomplete recovery or treatment resistance, for cases of emergency and for side effects associated with antipsychotic treatment. PMID- 12739996 TI - Therapeutic benefit of bisphosphonates in the management of prostate cancer related bone disease. AB - A diversity of bone pathology is present in men with prostate cancer. Androgen deprivation therapy (ADT) can cause significant and progressive osteopoenia and osteoporosis. Bone is also the primary site for metastases leading to associated pain, skeletal fractures and hypercalcaemia. Bisphosphonate therapy decreases bone resorption, which may prevent or reverse loss of bone mineral density. Both pamidronate and zoledronic acid have proven efficacy in preventing ADT-induced bone loss. In a randomised, placebo-controlled trial, in men with hormone refractory prostate cancer, there was a decreased incidence of skeletal- related adverse events in men receiving zoledronic acid. So far, randomised trials have failed to show improved pain control. Formalised guidelines are needed to help clinicians decide which patients should be treated with bisphosphonates, when to initiate therapy and for what duration. PMID- 12739998 TI - Atypical antipsychotics in the treatment of bipolar disorder. AB - Atypical antipsychotic medications are widely used for the treatment of bipolar disorder. Most empirical support suggests that these medications are efficacious in the treatment of acute mania, but there is considerably less support for the utility of these drugs in other phases of bipolar disorder. However, it is likely that several of these drugs will demonstrate efficacy in relapse prevention, and perhaps antidepressant efficacy in bipolar disorder as more studies are conducted. Atypical antipsychotics offer different side effect profiles than older antipsychotics, which may be of benefit for some patients. Consequently, atypical antipsychotics provide an important treatment option for bipolar patients. PMID- 12739999 TI - Paediatric community-acquired pneumonia: current concept in pharmacological control. AB - Community-acquired pneumonia (CAP) is one of the most frequent infections in childhood but it is not easy to establish a rational therapeutic approach for a number of reasons, including difficulties in identifying the aetiology, the fact that the most frequent bacterial pathogens become resistant to commonly used antibiotics and the lack of certain information concerning the possible preventive role of conjugate vaccines. This leads paediatricians to treat almost all cases of CAP with antibiotics, often using a combination of different antimicrobial classes. In order to avoid unnecessary antibiotic use and limit the spread of antibiotic resistance, consensus guidelines for the management of CAP in childhood should be developed and used by practitioners in their offices and hospitals. PMID- 12740000 TI - Colesevelam HCl: a non-systemic lipid-altering drug. AB - Colesevelam HCl (WelChol, Sankyo Pharmaceuticals Inc.) is a bile acid sequestrant polymer, which has been shown to significantly lower low density lipoprotein cholesterol and favourably affect high-density lipoprotein cholesterol blood levels in monotherapy and in combination with statins (HMG-CoA reductase inhibitors). Although it is similar to other bile acid sequestrants in that it binds bile acids and is non-systemic, colesevelam HCl differs in that it has a unique polymer structure that allows for greater tolerability with less potential drug interactions than with resins. Currently, statins are the most commonly prescribed lipid-altering drugs. However, it is not uncommon that patients demonstrate true or perceived intolerances to statin therapy, that are often dose related and may include elevations in liver or muscle enzyme blood levels, or myalgias or muscle weakness without muscle enzyme elevation. In rare circumstances, myopathy and even rhabdomyolysis can occur with statins. In addition, many statins also have important potential drug interactions. Finally, statin monotherapy is often not sufficient in achieving lipid treatment goals in many severely dyslipidaemic patients and the availability of colesevelam HCl provides a lipid-altering treatment addition to other lipid-altering drugs. From a clinical perspective, such combination therapy is often required to achieve treatment goals [1] in patients with more complicated or severe dyslipidaemia. Colesevelam HCl may also be an alternative in monotherapy for many patients with mild-to-moderate hypercholesterolaemia, as well as in some patients at potential risk from systemic exposure to alternative lipid-altering drugs (such as young children and fertile women). PMID- 12740001 TI - Safety and efficacy of tenecteplase in acute myocardial infarction. AB - The use of intravenous thrombolytic agents has revolutionised the treatment of acute myocardial infarction. However, the improved mortality achieved with these drugs is tempered by the risk of serious bleeding complications, especially intracranial haemorrhage (ICH). Tenecteplase (TNKase, Genetech Inc.) is an engineered variant of alteplase (Activase, Genentech Inc.) designed to have increased fibrin specificity, greater efficacy and a longer half-life. The longer half-life of tenecteplase compared to alteplase allows for convenient single bolus administration of the drug. In addition, tenecteplase dosing is based on actual or estimated patient weight, which enhances both the safety and efficacy outcomes. Large clinical trials have demonstrated equivalence in mortality and ICH between tenecteplase and alteplase. Compared to alteplase, tenecteplase use leads to lower rates of bleeding complications and a decreased risk of ICH among low weight, elderly women. PMID- 12740002 TI - Zanamivir in the treatment of influenza. AB - Influenza is a common illness, affecting many people every winter, with a considerable impact on mortality, hospital admissions, healthcare utilisation and sickness absence from work and school. Influenza management is currently focused on annual vaccinations for those in certain risk groups. Risk is determined by age and chronic illness, particularly diabetes, chronic respiratory and cardiac disease, and persons immunocompromised from disease or concomitant therapy. Amantadine (and in some countries, rimantadine is available but has not been widely used, because it is only effective against influenza A infections. The use of amantadines for treatment has been associated with the rapid emergence of resistant viruses capable of transmission, compromising its potential as a prophylactic, as well as a treatment. Side effects are well recognised and are a particular problem in the most vulnerable elderly populations, where dose restriction is necessary and prior knowledge of creatinine clearance desirable. The potential market for a new influenza treatment is large and the potential role of neuraminidase inhibitors in addressing this market has been covered in several review articles [1-4]. This review reports on the introduction of zanamivir (Relenza) to the market with particular reference to experience in the UK. PMID- 12740003 TI - Caspofungin: first approved agent in a new class of antifungals. AB - Caspofungin (Cancidas, Merck & Co. Inc.) is the first echinocandin antifungal agent to gain FDA-approval for use in the US. It has excellent clinical activity against Candida spp. and Aspergillus spp. but lacks significant activity against Cryptococcus neoformans. Caspofungin may have some activity against dimorphic fungi such as Histoplasma capsulatum and Coccidioides immitis, but no clinical data is available for treatment of these infections. Caspofungin has demonstrated poor activity against most filamentous fungi in vitro. Several clinical trials have demonstrated its efficacy in the treatment of oropharyngeal, oesophageal and invasive candidiasis, as well as invasive aspergillosis. As a result of caspofungin's unique mechanism of action, and the high morbidity and mortality of invasive fungal infections, there is considerable interest in using this new antifungal agent as part of a combination antifungal therapy. In vitro studies and small case series indicate that caspofungin does not appear to be antagonistic when combined with other antifungals, such as itraconazole, voriconazole or amphotericin B against Aspergillus spp. Caspofungin exerts concentration-dependent killing effects in many different in vitro and animal models of disseminated fungal infection. The usual daily dose is 50 mg/day i.v. following a 70 mg i.v. loading dose. However, higher caspofungin doses have been safely administered and up to 70 mg/day can be administered for patients who fail to respond to lower doses. Caspofungin has an excellent safety profile with reduced toxicities, compared to other licensed antifungal agents. Fever, thrombophlebitis, headache and liver enzyme elevations were the most common drug related side effects reported in clinical trials so far. Additional data are needed to document its safety in long-term use, and with higher doses in patients with invasive fungal infections. Caspofungin is a promising agent as first-line therapy for invasive candidiasis, and as salvage therapy for invasive aspergillosis. However, more clinical data are needed to define its role as primary therapy for invasive aspergillosis, and its role in combination antifungal therapy. PMID- 12740004 TI - ACE inhibitors versus diuretics: ALLHAT versus ANBP2. AB - The ALLHAT (The Antihypertensive and Lipid-Lowering treatment to prevent Heart Attack Trial) trial enrolled hypertensive patients with at least one additional risk factor for coronary heart disease (CHD) to a comparison of the diuretic chlorthalidone, the calcium channel blocker, amlodipine, and the angiotensin converting enzyme (ACE) inhibitor, lisinopril. Throughout the study, chlorthalidone decreased the systolic blood pressure to a slightly, but significantly greater extent (0.8-3.1 mmHg) than amlodipine or lisinopril. No significance differences were reported for amlodipine versus chlorthalidone or lisinopril versus chlorthalidone on the primary outcome of combined incidence of fatal CHD and nonfatal myocardial infarction. The findings of ALLHAT support the use of thiazide-type diuretics as first choice pharmacological therapy in at risk patients with hypertension. ANBP2 (The Second Australian National Blood Pressure Study) was also a comparison between diuretics (hydrochlorothiazide) and ACE inhibitors (enalapril) but was performed in older hypertensives that had few previous cardiovascular events. Diastolic blood pressure reduction was similar in both groups at all times. The risk of the primary outcome of all cardiovascular events or death from any cause was 11% lower in the ACE group than the diuretic group and the benefit was predominantly in men. Thus, ANBP2 suggests that in relatively healthy elderly hypertensive patients, ACE inhibitors should be preferred to diuretics. PMID- 12740005 TI - Evaluation of inducible promoters on the secretion of a ZZ-proinsulin fusion protein in Escherichia coli. AB - Four inducible promoters, uspA, uspB, lacUV5 and malK, were evaluated in the expression of the fusion protein ZZ-proinsulin by Escherichia coli. The aim was to select for their effects on the most appropriate expression system (promoter and culture medium) for secretion of ZZ-proinsulin to the periplasmic space and culture medium. All the expression vectors contained the RNase III cleavage site to ensure that the mRNA translation rate remained independent of 5'-untranslated regions thus making promoter strength comparisons more accurate. The highest ZZ proinsulin secretion yields were 6.2 mg/g of dry cell weight in the periplasmic space and 2.6 mg/g of dry cell weight in the culture medium using the malK promoter. It was also demonstrated that the use of M9 minimal medium favours secretion. PMID- 12740006 TI - Molecular studies of identification of genes for osteoporosis: the 2002 update. AB - We aim to give a comprehensive review, updated to 2002, of the most important and representative molecular genetic studies, performed mainly within the past decade, that aimed to identify the gene(s) involved in osteoporosis. Early reviews were largely confined to association studies in humans, but we review here, separately, the results of both association and linkage studies in humans, and quantitative trait locus (QTL) mapping in animal models. The main results of all the studies are tabulated for comparison and ease of reference, and to provide a comprehensive retrospective view of molecular genetics studies of osteoporosis. The most striking findings and the most representative studies are singled out for comment regarding the immediacy of their influence on present understanding of the genetics of osteoporosis and on the current status of genetic research in osteoporosis. This is particularly relevant for studies on the association of the vitamin D receptor (VDR) gene, for which there has been a large body of studies and reviews published. The format adopted by this review should be ideal for accommodating future new advances and studies in a fairly young field that is still developing rapidly. PMID- 12740007 TI - Acute (24 h) activation of peroxisome proliferator-activated receptor-alpha (PPARalpha) reverses high-fat feeding-induced insulin hypersecretion in vivo and in perifused pancreatic islets. AB - Abnormal depletion or accumulation of islet lipid may be important for the development of pancreatic beta cell failure. Long-term lipid sensing by beta cells may be co-ordinated via peroxisome proliferator-activated receptors (PPARs). We investigated whether PPARalpha activation in vivo for 24 h affects basal and glucose-stimulated insulin secretion in vivo after intravenous glucose administration and ex vivo in isolated perifused islets. Insulin secretion after intravenous glucose challenge was greatly increased by high-fat feeding (4 weeks) but glucose tolerance was minimally perturbed, demonstrating insulin hypersecretion compensated for insulin resistance. The effect of high-fat feeding to enhance glucose-stimulated insulin secretion was retained in perifused islets demonstrating a stable, long-term effect of high-fat feeding to potentiate islet glucose stimulus-secretion coupling. Treatment of high-fat-fed rats with WY14,643 for 24 h reversed insulin hypersecretion in vivo without impairing glucose tolerance, suggesting improved insulin action, and ex vivo in perfused islets. PPARalpha activation only affected hypersecretion of insulin since glucose stimulated insulin secretion was unaffected by WY14,643 treatment in vivo in control rats or in perifused islets from control rats. Our data demonstrate that activation of PPARalpha for 24 h can oppose insulin hypersecretion elicited by high-fat feeding via stable long-term effects exerted on islet function. PPARalpha could, therefore, participate in ameliorating abnormal glucose homeostasis and hyperinsulinaemia in dietary insulin resistance via modulation of islet function, extending the established requirement for PPARalpha for normal islet lipid homeostasis. PMID- 12740008 TI - Differential recruitment of the coactivator proteins CREB-binding protein and steroid receptor coactivator-1 to peroxisome proliferator-activated receptor gamma/9-cis-retinoic acid receptor heterodimers by ligands present in oxidized low-density lipoprotein. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) colocalizes with oxidized low-density lipoprotein (LDL) in foam cells in atherosclerotic lesions. We have explored a potential role of oxidized fatty acids in LDL as PPARgamma activators. LDL from patients suffering from intermittent claudication due to atherosclerosis was analyzed using HPLC and gas chromatography/mass spectrophotometry and found to contain 9-hydroxy and 13-hydroxyoctadecadienoic acid (9- and 13-HODE), as well as 5-hydroxy-, 12-hydroxy- and 15 hydroxyeicosatetraenoic acid (5-, 12- and 15-HETE respectively). PPARgamma was potently activated by 13(S)-HODE and 15(S)-HETE, as judged by transient transfection assays in macrophages or CV-1 cells. 5(S)- and 12(S)-HETE as well as 15-deoxy-Delta(12,14)-prostaglandin J(2) also activated PPARgamma but were less potent. Interestingly, the effect of the lipoxygenase products 13(S)-HODE and 15(S)-HETE as well as of the drug rosiglitazone were preferentially enhanced by the coactivator CREB-binding protein, whereas the effect of the cyclooxygenase product 15-deoxy-Delta(12,14)-prostaglandin J(2) was preferentially enhanced by steroid receptor coactivator-1. We interpret these results, which may have relevance to the pathogenesis of atherosclerosis, to indicate that the lipoxygenase products on the one hand and the cyclooxygenase product on the other exert specific effects on the transcription of target genes through differential coactivator recruitment by PPARgamma/9-cis retinoic acid receptor heterodimer complexes. PMID- 12740009 TI - Exogenous leptin controls the development of the small intestine in neonatal piglets. AB - Leptin, a hormone produced and secreted by adipose tIssue, muscles and stomach, is involved in the regulation of adipose tIssue mass, food intake and body weight in neonatal animals. It is also produced in the mammary glands and secreted into the colostrum and milk. Since leptin receptors are widely distributed in the small intestine mucosa, the aim of the present study was to investigate the effect of exogenous leptin on the development of the small intestine in neonatal piglets. Male neonatal piglets were fed with sow's milk or artificial milk formula. Every 8 h the latter received either vehicle or leptin (2 or 10 microg/kg body weight). The animals were either killed after 6 days of treatment and the small intestine sampled for histology and brush border enzyme activities or were tested for marker molecule (Na-fluorescein and BSA) absorption in vivo. Feeding milk formula slowed the maturation of small intestinal mucosa compared with feeding sow's milk. However, after leptin treatment the length of the small intestine was increased, and intestinal villi length, but not crypt size, was reduced compared with controls. The mitotic index was increased and the percentage of vacuolated enterocytes was reduced in the entire small intestine. Enterocyte brush border protease and lactase activities were reduced in the jejunum. Na-fluorescein marker molecule absorption did not change but that of BSA was reduced 3.8-fold. In conclusion, exogenous leptin administered in physiological doses reversed the maturation of the small intestinal mucosa to the range found in sow-reared piglets. PMID- 12740010 TI - Extrapituitary GH in the chicken: underestimation of immunohistochemical staining by Carnoy's fixation. AB - GH has previously been shown to be present in peripheral extrapituitary tIssues of chick embryos, but the cellular distribution of GH immunoreactivity is still uncertain because of differing immunohistochemical findings. The possibility that this uncertainty reflects differences in fixation of the embryonic tIssues was assessed by comparing GH immunoreactivity in tIssues fixed in 4% (w/v) paraformaldehyde or Carnoy's fluid (60% ethanol (v/v); 30% chloroform (v/v); 10% acetic acid (v/v)). A widespread distribution of GH immunoreactivity was seen in paraformaldehyde-fixed tIssues, although it was particularly intense in the spinal cord, dorsal and ventral root ganglia, notochord, myotome, epidermis, crop, heart, lung and humerus. In marked contrast, GH immunoreactivity in embryonic tIssues fixed with Carnoy's was more discrete and mainly restricted to marginal and mantle layers of the spinal cord, spinal nerves, the ventral root ganglia and the extensor nerve of the anterior limb bud. Since these are neural derivatives, Carnoy's fixation appears to preferentially result in neural GH staining, whereas GH staining in neural and non-neural tIssues is seen after paraformaldehyde fixation. Carnoy's, because it is a precipitive fixative, may only fix large GH moieties, whereas GH in peripheral tIssues includes numerous molecular variants, many of which are of relatively small size. Paraformaldehyde, because it is a cross-linking fixative, preferentially fixes peptides and small proteins, and it may therefore fix more GH moieties than Carnoy's fluid. Carnoy's fixation appears to underestimate GH immunoreactivity in immunohistochemical studies on the cellular distribution of GH-like proteins in embryonic chicks. PMID- 12740011 TI - Early growth restriction leads to down regulation of protein kinase C zeta and insulin resistance in skeletal muscle. AB - Epidemiological studies have revealed a relationship between early growth restriction and the subsequent development of type 2 diabetes. A rat model of maternal protein restriction has been used to investigate the mechanistic basis of this relationship. This model causes insulin resistance and diabetes in adult male offspring. The aim of the present study was to determine the effect of early growth restriction on muscle insulin action in late adult life. Rats were fed either a 20% or an isocaloric 8% protein diet during pregnancy and lactation. Offspring were weaned onto a 20% protein diet and studied at 15 Months of age. Soleus muscle from growth restricted offspring (LP) (of dams fed 8% protein diet) had similar basal glucose uptakes compared with the control group (mothers fed 20% protein diet). Insulin stimulated glucose uptake into control muscle but had no effect on LP muscle. This impaired insulin action was not related to changes in expression of either the insulin receptor or glucose transporter 4 (GLUT 4). However, LP muscle expressed significantly less (P<0.001) of the zeta isoform of protein kinase C (PKC zeta) compared with controls. This PKC isoform has been shown to be positively involved in GLUT 4-mediated glucose transport. Expression levels of other isoforms (betaI, betaII, epsilon, theta) of PKC were similar in both groups. These results suggest that maternal protein restriction leads to muscle insulin resistance. Reduced expression of PKC zeta may contribute to the mechanistic basis of this resistance. PMID- 12740012 TI - Expression of islet neogenesis-associated protein in islets of normal hamsters. AB - The aim of the present study was to test the possible presence and expression of islet neogenesis-associated protein (INGAP) in islet cells of normal adult hamsters. Pancreata from normal male Syrian hamsters were removed to perform the following studies. (i) Western blot analysis using the cytosolic fraction from homogenates of isolated islets, exocrine tIssue and whole pancreas, and rabbit INGAP-specific antibody. (ii) Immunohistochemical identification of INGAP positive cells in fixed sections of intact pancreata, fresh and 72 h cultured islets (isolated by collagenase digestion), and smears of exocrine pancreatic cells, using the same INGAP-specific antibody and streptavidin-biotin complex. (iii) RT-PCR using total RNA extracted from isolated islets and from exocrine tIssue as template, and a specific pair of primers. (iv) Control of the sequence of the PCR products. INGAP protein was identified by Western blot in the cytosolic fraction of homogenates from fresh isolated islets, exocrine cells and whole fresh pancreas. INGAP-immunopositive cells were observed in duct, exocrine and islet cells in either fixed intact or digested pancreatic tIssue. INGAP mRNA was identified in samples of total RNA from fresh and cultured isolated islets and from exocrine cells. Our data demonstrate that INGAP is present and expressed in islets and in exocrine pancreatic cells of normal hamsters. The ubiquitous localization of INGAP suggests its possible role in the physiological process of islet growth and its protective effect upon streptozotocin-induced diabetes. PMID- 12740013 TI - Pancreatic duodenal homeobox-1 and islet neogenesis-associated protein: a possible combined marker of activateable pancreatic cell precursors. AB - The aim of this work was to study the possible relationship between pancreatic duodenal homeobox-1 (Pdx-1) and islet neogenesis-associated protein (INGAP) during induced islet neogenesis. Pregnant hamsters were fed with (S) and without (C) sucrose, and glycemia, insulin secretion in vitro, and pancreas immunomorphometric parameters were measured in their 7-day-old offspring. S offspring had significantly lower glycemic levels than C animals. Insulin release in response to increasing glucose concentrations in the incubation medium (2-16 mM glucose) did not increase in pancreata from either C or S offspring. However, pancreata from S offspring released more insulin than those from C animals. In S offspring, beta-cell mass, beta-cell replication rate and islet neogenesis increased significantly, with a simultaneous decrease in beta-cell apoptotic rate. INGAP- and Pdx-1-positive cell mass also increased in the islets and among acinar and duct cells. We found two subpopulations of Pdx-1 cells: INGAP-positive and INGAP-negative. Pdx-1/INGAP-positive cells did not stain with insulin, glucagon, somatostatin, pancreatic polypeptide, or neurogenin 3 antibodies. The increment of Pdx-1/INGAP-positive cells represented the major contribution to the Pdx-1 cell mass increase. Such increments varied among pancreas subsectors: ductal>insular>extrainsular. Our results suggested that INGAP participates in the regulation of islet neogenesis, and Pdx-1/INGAP-positive cells represent a new stem cell subpopulation at an early stage of development, highly activateable in neogenesis. PMID- 12740014 TI - Increased 5'-iodothyronine deiodinase activity is a maternal adaptive mechanism in response to protein restriction during lactation. AB - We have shown that protein restriction during lactation is associated with higher levels of serum and milk tri-iodothyronine (T(3)) with lower serum thyroxine (T(4)), suggesting an increased T(4) to T(3) conversion. To investigate this hypothesis, the activity of type 1 (D1) and/or type 2 (D2) iodothyronine deiodinases was evaluated on days 4, 12 and 21 of lactation in several tIssues of dams fed an 8% protein-restricted (PR) diet and controls fed a 23% protein diet. Serum TSH, T(3) and T(4) were measured by radioimmunoassay. Deiodinase activity was determined by the release of (125)I from (125)I-reverse T(3), under specific conditions for D1 or D2. PR dams had a transitory reduction in liver D1 activity (P<0.05) on day 12, and a small increase in thyroid D1 on day 12 followed by a small decrease on day 21. However, thyroid D2 activity was higher than controls (P<0.05) during the whole of the lactation period. Mammary gland D1 and D2 activities were lower on day 4 of lactation in PR dams (P<0.05), and D2 was higher on day 21 (P<0.05). Potentially, a lower conversion of T(3) to di iodothyronine in the mammary glands of PR dams at the beginning of lactation may serve to provide more T(3) through the milk. Brown adipose tIssue (BAT) D2 activity was higher (P<0.05) in PR dams during all periods of lactation. PR dams showed higher skeletal muscle D1 activity only at the end of lactation, but no changes in D2 activity. Higher pituitary D1 and D2 activities in the PR group (P<0.05) at the end of lactation could have contributed to the lower serum TSH. These data suggest that the higher thyroid and BAT D2 activity during the whole of lactation and skeletal muscle D1 activity at the end of lactation may contribute to the higher serum T(3) in PR dams. PMID- 12740015 TI - Relationships between cell division, expression of growth factors and microcirculation in the thyroids of Tg-A2aR transgenic mice and patients with Graves' disease. AB - Tissue heterogeneity and nodule formation are hallmarks of thyroid growth. This is accounted for by the clonality theory that acknowledges different individual cellular abilities to respond to trophic stimuli. In order to test the hypothesis that functional and mitotic properties of thyrocytes could be influenced by paracrine interactions with neighbour endothelial cells, studies were conducted in both mouse and human goitre models. In the first part of the study, homogenous goitres in C57 black mice were compared with heterogeneous goitres in transgenic hyperthyroid mice expressing the A2 adenosine receptor (Tg-A2aR). The second part of the study concentrated on comparing human thyroid tIssue of control individuals and of patients with Graves' disease. The rate of cell division was evaluated by immunohistochemical detection of cells positive for proliferating cell nuclear antigen (PCNA). Their spatial distribution was then correlated with immunohistochemical cellular expression of growth- and vasoactive-related factors (fibroblast growth factor-2, transforming growth factor-beta, endothelin-1, vascular endothelial growth factor, nitric oxide synthase III), and with microcirculation expansion. Observations were made on digitalised images of histological serial sections. The nearest-neighbour method was used to distinguish between random or clustered distribution. PCNA-positive cells were both randomly and uniformly distributed in homogenous goitres from C57 black mice, and were clustered in tIssue areas identified as papillary and hyperplastic zones in heterogeneous goitres from Tg-A2aR mice. However, they were absent in the so-called compact cellular zones featuring resting cells. Moreover, whereas papillary and hyperplastic zones were highly vascularised, compact zones were nearly free of microvessels. Spatial distribution of dividing cells was positively correlated with the expression of growth-related factors. A similar pattern was observed in the thyroids of patients with Graves' disease. In accordance with the recent demonstration of the presence of angiofollicular units in the thyroid, these data strongly support the hypothesis that functional and mitotic properties of each single thyrocyte, likely to be responsible for growth heterogeneity of hyperplastic glands, may be adjusted at tIssue level by specific interactions with neighbour endothelial cells that, in turn, could alter the mitotic rate of thyrocytes through paracrine signals. PMID- 12740016 TI - Identification of somatostatin receptors controlling growth hormone and thyrotropin secretion in the chicken using receptor subtype-specific agonists. AB - Somatostatin (SRIH) functions as an endocrine mediator in processes such as growth, immune resistance and reproduction. Five SRIH receptors (sstr1-5) have been identified in mammals, where they are expressed in both the brain and peripheral tIssues. To study the specific function of each receptor subtype, specific agonists (ag1-5) have been synthesized. The high degree of homology between mammalian and avian SRIH receptors suggests that these agonists might also be used in chickens. In this paper we describe two in vitro protocols (static incubation and perifusion system) to identify the SRIH receptors controlling the secretion of GH and TSH from the chicken pituitary. We found that basal GH or TSH secretion were never affected when SRIH or an agonist (1 microM) were added. SRIH diminished the GH as well as the TSH response to TSH-releasing hormone (TRH; 100 nM) in both systems. Our results have indicated that the SRIH actions at the level of the pituitary are regulated through specific receptor subtypes. In both the static and flow incubations, ag2 lowered the GH response to TRH, whereas stimulated TSH release was diminished by both ag2 and ag5. Ag3 and ag4 tended to increase rather than decrease the responsiveness of both pituitary cell types to TRH in perifusion studies. Our data have indicated that SRIH inhibits chicken pituitary function through sstr2 and sstr5. Only sstr2 seems to be involved in the control of chicken GH release, whereas both sstr2 and sstr5 inhibit induced GH secretion in mammals. The possible stimulatory action of ag3 and ag4 may point towards a species-specific function of sstr3 and sstr4. PMID- 12740017 TI - Ovarian and hormonal response of female goats to active immunization against inhibin. AB - This study was conducted to evaluate the effect of active immunization against inhibin on hormonal levels and the ovulation rate in goats. Ten adult Shiba goats (Capra hircus) in two groups were used in this study. The first group was injected with inhibin vaccine (immunized, n=5) and the second group was injected with Freund's adjuvant (control, n=5) followed by three booster injections at 4 week intervals. After the third booster injection, three consecutive periods of oestrus were induced using prostaglandin F(2alpha) at intervals of 11 days. Blood samples were collected at 2-6 h intervals and the ovaries were monitored using B mode ultrasonography. All inhibin-immunized goats generated antibodies that bound (125)I-labelled bovine inhibin and their FSH concentrations were significantly higher than corresponding values in the control group. Also, inhibin-immunized goats had significantly higher preovulatory oestradiol-17beta (P<0.01) and higher concentrations of progesterone in the luteal phase (P<0.05). Immunization of goats against inhibin resulted in a significant (P<0.01) increase in ovulation rate (control: 1.7+/-0.3 vs immunized: 7.6+/-1.1). These results demonstrate that active immunization against inhibin enhances ovarian follicular development and ovulation rate by promoting an increase in pituitary FSH secretion. Therefore, immunization against inhibin may be a useful alternative to the conventional approach of superovulation in goats. PMID- 12740018 TI - Interactions between the ovary and the local IGF-I axis modulate mammary development in prepubertal heifers. AB - The objective was to determine the effects of ovariectomy and epithelial-stromal interactions on mammary development and local expression of IGF-I and IGF-binding protein (IGFBP) mRNA in prepubertal heifers. An epithelium-free ('cleared') fat pad (CFP) was prepared in two glands in each of 14 Holstein heifers, aged 1-3 Months. Eight of the calves were also ovariectomized. Serum concentrations of GH, IGF-I and prolactin were not affected by ovariectomy. At 6 Months of age, calves were killed to provide mammary samples of parenchyma, CFP and intact fat pad (MFP). Total mammary mass was reduced in ovariectomized calves (130+/-21 g vs 304+/- 25 g; P<0.001), and in several cases parenchymal tIssue was essentially absent. Uterus weight was also reduced by ovariectomy (14.5+/-3.8 g vs 30.4+/-4.5 g; P<0.05). In support of our hypothesis that local IGF-I mediates prepubertal mammary development, mRNA expression of IGF-I was lower in ovariectomized than in control calves (62.1+/-7.8 vs 91.6+/-7.8 arbitrary units; P<0.05). Specific binding of IGF-I to mammary parenchymal microsomes was also reduced by ovariectomy (377+/-142 vs 868+/-82 c.p.m.; P<0.01), suggesting decreased sensitivity to IGF-I. Expression of IGFBP-3 and IGFBP-5 mRNA were not influenced by ovariectomy. Expression of IGF-I, IGFBP-3 and IGFBP-5 mRNA did not differ between CFP and MFP, suggesting that expression of these factors was not influenced by interactions between stroma and developing epithelium. Overall, the data suggested that interactions between the ovary and the local IGF-I axis act to optimize the availability and effectiveness of IGF-I within the gland to stimulate mammary growth. PMID- 12740020 TI - Localization of estrogen receptors-alpha and -beta and androgen receptor in the human growth plate at different pubertal stages. AB - Sex steroids are required for a normal pubertal growth spurt and fusion of the human epiphyseal growth plate. However, the localization of sex steroid receptors in the human pubertal growth plate remains controversial. We have investigated the expression of estrogen receptor (ER) alpha, ERbeta and androgen receptor (AR) in biopsies of proximal tibial growth plates obtained during epiphyseal surgery in 16 boys and eight girls. All pubertal stages were represented (Tanner stages 1 5). ERalpha, ERbeta and AR were visualized with immunohistochemistry and the number of receptor-positive cells was counted using an image analysis system. Percent receptor-positive chondrocytes were assessed in the resting, proliferative and hypertrophic zones and evaluated for sex differences and pubertal trends. Both ERalpha- and ERbeta-positive cells were detected at a greater frequency in the resting and proliferative zones than in the hypertrophic zone (64+/-2%, 64+/-2% compared with 38+/-3% for ERalpha, and 63+/-3%, 66+/-3% compared with 53+/-3% for ERbeta), whereas AR was more abundant in the resting (65+/-3%) and hypertrophic zones (58+/-3%) than in the proliferative zone (41+/ 3%). No sex difference in the patterns of expression was detected. For ERalpha and AR, the percentage of receptor-positive cells was similar at all Tanner pubertal stages, whereas ERbeta showed a slight decrease in the proliferative zone during pubertal development (P<0.05). In summary, our findings suggest that ERalpha, ERbeta and AR are expressed in the human growth plate throughout pubertal development, with no difference between the sexes. PMID- 12740019 TI - Expression and localisation of oestrogen and progesterone receptors in the bovine mammary gland during development, function and involution. AB - It is now well established that oestrogen and progesterone are absolutely essential for mammary gland development. Lactation can be induced in non-pregnant animals by sex steroid hormone treatment. Most of the genomic actions of oestrogens are mediated by two oestrogen receptors (ER)-alpha and ERbeta, and for gestagens in ruminants by the progesterone receptor (PR). Our aim was the evaluation of mRNA expression and protein (localisation and Western blotting) during mammogenesis, lactogenesis, galactopoiesis (early, middle and late) and involution (8, 24, 28, 96-108 h and 14-28 days after the end of milking) in the bovine mammary gland (total no. 53). During these stages, the mRNA was assessed by means of real-time RT-PCR (LightCycler). The protein for ERalpha, ERbeta and PR was localised by immunohistochemistry and Western blotting. The mRNA expression results indicated the existence of ERalpha, ERbeta and PR in bovine mammary gland. Both ERalpha and PR are expressed in fg/ micro g total RNA range. The highest mRNA expression was found for ERalpha and PR in the tIssue of non pregnant heifers, followed by a significant decrease to a lower level at the time of lactogenesis with low concentrations remaining during lactation and the first 4 weeks of involution. In contrast, the expression of ERbeta was about 1000-fold lower (ag/ micro g total RNA) and showed no clear difference during the stages examined, with a significant increase only 2-4 weeks after the end of milking. Immunolocalisation for ERalpha revealed a strong positive staining in nuclei of lactocytes in non-pregnant heifers, became undetectable during pregnancy, lactogenesis and lactation, and was again detectable 14-28 days after the end of milking. In contrast, PR was localised in the nuclei of epithelial cells in the mammary tIssue of non-pregnant heifers, in primigravid animals, and during late lactation and involution. During lactogenesis, peak and mid lactation, fewer nuclei of epithelial cells were positive, but increased staining of the cytoplasm of epithelial cells was obvious. ERalpha and ERbeta protein was found in all mammary gland stages examined by Western blotting. In contrast to mRNA expression, the protein signal for ERalpha was weaker in the tIssue of non pregnant heifers and during involution (4 weeks). ERbeta protein showed a stronger signal (two isoform bands) in non-pregnant heifers and 4 weeks after the end of milking. This correlated with the mRNA expression data. Three isoforms of PR (A, B and C) were found by Western blotting in the tIssue of non-pregnant heifers, but only isoform B remained during the following stages (lactogenesis, galactopoiesis and involution). In conclusion, the mRNA expression and protein data for ER and PR showed clear regulatory changes, suggesting involvement of these receptors in bovine mammary gland development and involution. PMID- 12740021 TI - Modulation of gap junction mediated intercellular communication in TM3 Leydig cells. AB - Long-term modulation of intercellular communication via gap junctions was investigated in TM3 Leydig cells, under low and high confluence states, and upon treatment of the cells for different times with activators of protein kinase A (PKA) and protein kinase C (PKC). Cells in low confluence were readily coupled, as determined by transfer of the dye Lucifer Yellow; on reaching confluence, the cells uncoupled. Western blots and RT-PCR revealed that connexin 43 (Cx43) was abundantly expressed in TM3 Leydig cells and its expression was decreased after the cells achieved confluence. Stimulation of PKA or PKC induced a decrease in cell-cell communication. Staurosporin, an inhibitor of protein kinases, increased coupling and was able to prevent and reverse the uncoupling actions of dibutyryl cAMP and 12-O-tetradecanoyl-phorbol-13-acetate (TPA). Under modulation by confluence, Cx43 was localized to the appositional membranes when cells were coupled and was mainly in the cytoplasm when they were uncoupled. In addition, cAMP and TPA reduced the surface membrane labeling for Cx43, whereas staurosporin increased it. These data show a strong correlation between functional coupling and the membrane distribution of Cx43, implying that this connexin has an important role in intercellular communication between TM3 cells. Furthermore, increased testosterone secretion in response to luteinizing hormone was accompanied by a decrease in intercellular communication, suggesting that gap junction mediated coupling may be a modulator of hormone secretion in TM3 cells. PMID- 12740022 TI - A comparison between three commercial kits for chromogranin A measurements. AB - Chromogranin (CgA) has been shown to be an excellent marker for neuroendocrine tumours. There are now three commercial assays on the market. We wanted to compare the usefulness of the different kits in a clinical situation. We have thus measured CgA in 77 patients and compared the results from the different methods. CgA was measured with three different commercial kits according to the recommendations from the manufacturers (CGA-RIA CT; CIS bio international, Gif sur-Yvette cedex, France, DAKO Chromogranin A ELISA kit; DAKO A/S, Glostrup, Denmark and CgA; EuroDiagnostica, Malmo, Sweden). The sensitivity and specificity differed between the different kits. The CIS kit showed a sensitivity of 67% and a specificity of 96%. The sensitivity and specificity were both 85% for the DAKO kit and 93% and 88% respectively for the EuroDiagnostica assay. We have concluded that CgA is an important tumour marker for all neuroendocrine tumours. However, different analytical properties of the respective kits give different performances, a fact that must be taken into consideration when comparing results from different clinical studies. A recognised international standard for CgA would be a step on the way to harmonisation, but antibody selection and construction of the assays will probably still influence the results. PMID- 12740023 TI - Risk factors of thyroid abnormalities in bipolar patients receiving lithium: a case control study. AB - BACKGROUND: Lithium-induced thyroid abnormalities have been documented in many studies. They may occur despite normal plasma lithium levels. The objectives of this study were: 1) to determine possible relationship between lithium ratio, defined as erythrocyte lithium concentrations divided by plasma lithium concentrations, and thyroid abnormalities in bipolar patients receiving lithium and 2) to find other possible risk factors for developing thyroid abnormalities in the subjects. METHODS: Sixty-eight bipolar patients receiving lithium therapy were enrolled in a cross-sectional evaluation of thyroid function test and thyroid size. Patients were divided into two groups based on their thyroid function tests and thyroid sizes. Erythrocyte and plasma lithium concentrations were determined by atomic absorption spectrometry for each patient. Lithium ratio was then calculated. RESULTS: No significant differences were found between age, positive family history of affective disorder, plasma lithium concentration, erythrocyte lithium concentration, and lithium ratio comparing the two groups. Thyroid abnormalities was significantly higher in women than in men (p < 0.05). CONCLUSIONS: Lithium ratio does not appear to have a predictive role for thyroidal side effects of lithium therapy. Female gender was the main risk factor. We suggest more frequent thyroid evaluation of bipolar women who are treated with lithium. PMID- 12740024 TI - The Canadian celiac health survey--the Ottawa chapter pilot. AB - BACKGROUND: Celiac disease may manifest with a variety of symptoms which can result in delays in diagnosis. Celiac disease is associated with a number of other medical conditions. The last national survey of members of the Canadian Celiac Association (CCA) was in 1989. Our objective was to determine the feasibility of surveying over 5,000 members of the CCA, in addition to obtaining more health related information about celiac disease. METHODS: The Professional Advisory Board of the CCA in collaboration with the University of Ottawa developed a comprehensive questionnaire on celiac disease. The questionnaire was pre-tested and then a pilot survey was conducted on members of the Ottawa Chapter of the CCA using a Modified Dillmans' Total Design method for mail surveys. RESULTS: We had a 76% response to the first mailout of the questionnaire. The mean age of participants was 55.5 years and the mean age at diagnosis was 45 years. The majority of respondents presented with abdominal pain, diarrhea, fatigue or weight loss. Prior to diagnosis, 30% of respondents consulted four or more family doctors. Thirty seven percent of individuals were told they had either osteoporosis or osteopenia. Regarding the impact of the gluten-free diet (GFD), 45% of individuals reported that they found following a GFD was very or moderately difficult. The quality of life of individuals with celiac disease was comparable to the mean quality of life of Canadians. CONCLUSION: On the basis of our results, we concluded that a nationwide survey is feasible and this is in progress. Important concerns included delays in the diagnosis of celiac disease and the awareness of associated medical conditions. Other issues include awareness of celiac disease by health professionals and the impact of the GFD on quality of life. These issues will be addressed further in the national survey. PMID- 12740025 TI - Evidence-based medicine in primary care: qualitative study of family physicians. AB - BACKGROUND: The objectives of this study were: a) to examine physician attitudes to and experience of the practice of evidence-based medicine (EBM) in primary care; b) to investigate the influence of patient preferences on clinical decision making; and c) to explore the role of intuition in family practice. METHOD: Qualitative analysis of semi-structured interviews of 15 family physicians purposively selected from respondents to a national survey on EBM mailed to a random sample of Canadian family physicians. RESULTS: Participants mainly welcomed the promotion of EBM in the primary care setting. A significant number of barriers and limitations to the implementation of EBM were identified. EBM is perceived by some physicians as a devaluation of the 'art of medicine' and a threat to their professional/clinical autonomy. Issues regarding the trustworthiness and credibility of evidence were of great concern, especially with respect to the influence of the pharmaceutical industry. Attempts to become more evidence-based often result in the experience of conflicts. Patient factors exert a powerful influence on clinical decision-making and can serve as trumps to research evidence. A widespread belief that intuition plays a vital role in primary care reinforced views that research evidence must be considered alongside other factors such as patient preferences and the clinical judgement and experience of the physician. DISCUSSION: Primary care physicians are increasingly keen to consider research evidence in clinical decision-making, but there are significant concerns about the current model of EBM. Our findings support the proposed revisions to EBM wherein greater emphasis is placed on clinical expertise and patient preferences, both of which remain powerful influences on physician behaviour. PMID- 12740027 TI - Genomic homogeneity between Mycobacterium avium subsp. avium and Mycobacterium avium subsp. paratuberculosis belies their divergent growth rates. AB - BACKGROUND: Mycobacterium avium subspecies avium (M. avium) is frequently encountered in the environment, but also causes infections in animals and immunocompromised patients. In contrast, Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) is a slow-growing organism that is the causative agent of Johne's disease in cattle and chronic granulomatous infections in a variety of other ruminant hosts. Yet we show that despite their divergent phenotypes and the diseases they present, the genomes of M. avium and M. paratuberculosis share greater than 97% nucleotide identity over large (25 kb) genomic regions analyzed in this study. RESULTS: To characterize genome similarity between these two subspecies as well as attempt to understand their different growth rates, we designed oligonucleotide primers from M. avium sequence to amplify 15 minimally overlapping fragments of M. paratuberculosis genomic DNA encompassing the chromosomal origin of replication. These strategies resulted in the successful amplification and sequencing of a contiguous 11-kb fragment containing the putative Mycobacterium paratuberculosis origin of replication (oriC). This fragment contained 11 predicted open reading frames that showed a conserved gene order in the oriC locus when compared with several other Gram-positive bacteria. In addition, a GC skew analysis identified the origin of chromosomal replication which lies between the genes dnaA and dnaN. The presence of multiple DnaA boxes and the ATP-binding site in dnaA were also found in M. paratuberculosis. The strong nucleotide identity of M. avium and M. paratuberculosis in the region surrounding the origin of chromosomal replication led us to compare other areas of these genomes. A DNA homology matrix of 2 million nucleotides from each genome revealed strong synteny with only a few sequences present in one genome but absent in the other. Finally, the 16s rRNA gene from these two subspecies is 100% identical. CONCLUSIONS: We present for the first time, a description of the oriC region in M. paratuberculosis. In addition, genomic comparisons between these two mycobacterial subspecies suggest that differences in the oriC region may not be significant enough to account for the diverse bacterial replication rates. Finally, the few genetic differences present outside the origin of chromosomal replication in each genome may be responsible for the diverse growth rates or phenotypes observed between the avium and paratuberculosis subspecies. PMID- 12740026 TI - Profilin is required for viral morphogenesis, syncytium formation, and cell specific stress fiber induction by respiratory syncytial virus. AB - BACKGROUND: Actin is required for the gene expression and morphogenesis of respiratory syncytial virus (RSV), a clinically important Pneumovirus of the Paramyxoviridae family. In HEp-2 cells, RSV infection also induces actin stress fibers, which may be important in the immunopathology of the RSV disease. Profilin, a major regulator of actin polymerization, stimulates viral transcription in vitro. Thus, we tested the role of profilin in RSV growth and RSV-actin interactions in cultured cells (ex vivo). RESULTS: We tested three cell lines: HEp-2 (human), A549 (human), and L2 (rat). In all three, RSV grew well and produced fused cells (syncytium), and two RSV proteins, namely, the phosphoprotein P and the nucleocapsid protein N, associated with profilin. In contrast, induction of actin stress fibers by RSV occurred in HEp-2 and L2 cells, but not in A549. Knockdown of profilin by RNA interference had a small effect on viral macromolecule synthesis but strongly inhibited maturation of progeny virions, cell fusion, and induction of stress fibers. CONCLUSIONS: Profilin plays a cardinal role in RSV-mediated cell fusion and viral maturation. In contrast, interaction of profilin with the viral transcriptional proteins P and N may only nominally activate viral RNA-dependent RNA polymerase. Stress fiber formation is a cell-specific response to infection, requiring profilin and perhaps other signaling molecules that are absent in certain cell lines. Stress fibers per se play no role in RSV replication in cell culture. Clearly, the cellular architecture controls multiple steps of host-RSV interaction, some of which are regulated by profilin. PMID- 12740028 TI - Development and validation of a T7 based linear amplification for genomic DNA. AB - BACKGROUND: Genomic maps of transcription factor binding sites and histone modification patterns provide unique insight into the nature of gene regulatory networks and chromatin structure. These systematic studies use microarrays to analyze the composition of DNA isolated by chromatin immunoprecipitation. To obtain quantities sufficient for microarray analysis, the isolated DNA must be amplified. Current protocols use PCR-based approaches to amplify in exponential fashion. However, exponential amplification protocols are highly susceptible to bias. Linear amplification strategies minimize amplification bias and have had a profound impact on mRNA expression analysis. These protocols have yet to be applied to the analysis of genomic DNA due to the lack of a suitable tag such as the polyA tail. RESULTS: We have developed a novel linear amplification protocol for genomic DNA. Terminal transferase is used to add polyT tails to the ends of DNA fragments. Tail length uniformity is ensured by including a limiting concentration of the terminating nucleotide ddCTP. Second strand synthesis using a T7-polyA primer adapter yields double stranded templates suitable for in vitro transcription (IVT). Using this approach, we are able to amplify as little as 2.5 ng of genomic DNA, while retaining the size distribution of the starting material. In contrast, we find that PCR amplification is biased towards species of greater size. Furthermore, extensive microarray-based analyses reveal that our linear amplification protocol preserves dynamic range and species representation more effectively than a commonly used PCR-based approach. CONCLUSION: We present a T7-based linear amplification protocol for genomic DNA. Validation studies and comparisons with existing methods suggest that incorporation of this protocol will reduce amplification bias in genome mapping experiments. PMID- 12740029 TI - A comparison of Pfam and MEROPS: two databases, one comprehensive, and one specialised. AB - BACKGROUND: We wished to compare two databases based on sequence similarity: one that aims to be comprehensive in its coverage of known sequences, and one that specialises in a relatively small subset of known sequences. One of the motivations behind this study was quality control. Pfam is a comprehensive collection of alignments and hidden Markov models representing families of proteins and domains. MEROPS is a catalogue and classification of enzymes with proteolytic activity (peptidases or proteases). These secondary databases are used by researchers worldwide, yet their contents are not peer reviewed. Therefore, we hoped that a systematic comparison of the contents of Pfam and MEROPS would highlight missing members and false-positives leading to improvements in quality of both databases. An additional reason for carrying out this study was to explore the extent of consensus in the definition of a protein family. RESULTS: About half (89 out of 174) of the peptidase families in MEROPS overlapped single Pfam families. A further 32 MEROPS families overlapped multiple Pfam families. Where possible, new Pfam families were built to represent most of the MEROPS families that did not overlap Pfam. When comparing the numbers of sequences found in the overlap between a MEROPS family and its corresponding Pfam family, in most cases the overlap was substantial (52 pairs of MEROPS and Pfam families had an intersection size of greater than 75% of the union) but there were some differences in the sets of sequences included in the MEROPS families versus the overlapping Pfam families. CONCLUSIONS: A number of the discrepancies between MEROPS families and their corresponding Pfam families arose from differences in the aims and philosophies of the two databases. Examination of some of the discrepancies highlighted additional members of families, which have subsequently been added in both Pfam and MEROPS. This has led to improvements in the quality of both databases. Overall there was a great deal of consensus between the databases in definitions of a protein family. PMID- 12740030 TI - Lysophosphatidic acid, LPA: a bad boy becomes good. PMID- 12740031 TI - Placental expression of estrogen receptor beta and its hormone binding variant- comparison with estrogen receptor alpha and a role for estrogen receptors in asymmetric division and differentiation of estrogen-dependent cells. AB - During human pregnancy, the production of 17-beta-estradiol (E2) rises steadily to eighty fold at term, and placenta has been found to specifically bind estrogens. We have recently demonstrated the expression of estrogen receptor alpha (ER-alpha) protein in human placenta and its localization in villous cytotrophoblast (CT), vascular pericytes, and amniotic fibroblasts. In vitro, E2 stimulated development of large syncytiotrophoblast (ST) aggregates. In the present study we utilized ER-beta affinity purified polyclonal (N19:sc6820) and ER-alpha monoclonal (clone h-151) antibodies. Western blot analysis revealed a single approximately 52 kDa ER-beta band in chorionic villi (CV) protein extracts. In CV, strong cytoplasmic ER-beta immunoreactivity was confined to ST. Dual color immunohistochemistry revealed asymmetric segregation of ER-alpha in dividing villous CT cells. Prior to separation, the cell nuclei more distant from ST exhibited high ER-alpha, while cell nuclei associated with ST showed diminution of ER-alpha and appearance of ER-beta. In trophoblast cultures, development of ST aggregates was associated with diminution of ER-alpha and appearance of ER-beta immunoreactivity. ER-beta was also detected in endothelial cells, amniotic epithelial cells and fibroblasts, extravillous trophoblast (nuclear and cytoplasmic) and decidual cells (cytoplasmic only). In addition, CFK E12 (E12) and CWK-F12 (F12) monoclonal antibodies, which recognize approximately 64 kDa ER-beta with hormone binding domain, showed nuclear-specific reactivity with villous ST, extravillous trophoblast, and amniotic epithelium and fibroblasts. Western blot analysis indicated abundant expression of a approximately 64 kDa ER-beta variant in trophoblast cultures, significantly higher when compared to the chorionic villi and freshly isolated trophoblast cell protein extracts. This is the first report on ER-beta expression in human placenta and cultured trophoblast. Our data indicate that during trophoblast differentiation, the ER-alpha is associated with a less, and ER-beta with the more differentiated state. Enhanced expression of approximately 64 kDa ER-beta variant in trophoblast cultures suggests a unique role of ER-beta hormone binding domain in the regulation of trophoblast differentiation. Our data also indicate that asymmetric segregation of ER-alpha may play a role in asymmetric division of estrogen-dependent cells. PMID- 12740032 TI - IL-11 and IL-11Ralpha immunolocalisation at primate implantation sites supports a role for IL-11 in placentation and fetal development. AB - Embryo implantation, endometrial stromal cell decidualization and formation of a functional placenta are critical processes in the establishment and maintenance of pregnancy. Interleukin (IL)-11 signalling is essential for adequate decidualization in the mouse uterus and IL-11 promotes decidualization in the human. IL-11 action is mediated via binding to the specific IL-11 receptor alpha (IL-11Ralpha). The present study examined immunoreactive IL-11 and IL-11Ralpha in cycling rhesus monkey endometrium, at implantation sites in cynomolgus and rhesus monkeys and in human first trimester decidua and defined distinct spatial and temporal patterns. In cycling rhesus monkey endometrium, IL-11 and IL-11Ralpha increased in both basalis and functionalis regions during the secretory compared with the proliferative phase, with changing cellular locations in luminal and glandular epithelium and stroma. The patterns were similar overall to those previously described in human endometrium. Differences were seen in immunostaining during implantation in cynomologus and rhesus monkey. In the cynomolgus, very little staining for IL-11 or IL-11Ralpha was seen in syncytio- and cyto-trophoblast cells in the villi between days 12 and 150 of pregnancy although there was moderate staining in cytotrophoblast in the shell between days 12 and 17 and in subpopulations of cytotrophoblast cells invading the arteries at day 17. By contrast in the rhesus monkey between days 24 and 35 of pregnancy and in human first trimester placenta, cyto- and syncytio-trophoblast in the villi but not cytotrophoblast in the shell were positively stained. The most intense staining for both IL-11 and IL-11Ralpha was present within the decidua in the maternal component of implantation sites in all three primates but moderate staining was also present in maternal vascular smooth muscle and glands perivascular cells and epithelial plaques. These results are consistent with a role for IL-11 both during decidualization and placentation in primates. PMID- 12740034 TI - Dyspnea and quality of life indicators in hospice patients and their caregivers. AB - This study describe the assessment of dyspnea, symptom distress, and quality of life measures in 163 hospice patients with cancer who reported dyspnea. Mean age of the hospice patient sample was 70.22 years and 61.86 for caregivers (65% were spouses). The majority of patients and caregivers were white: 87%, 63% of the patients were male while 78% of caregivers were female. Mean dyspnea intensity as reported by patients was 4.52 (SD 2.29) and caregivers, 4.39 (SD 2.93). Patients' and caregivers' ratings of the patient's dyspnea intensity revealed no significant differences in ratings thus verifying that caregivers can assess dyspnea severity accurately. Patients' perceived quality of life ratings were not significantly correlated with ratings of their caregivers' perceived quality of life. For patients, symptom distress and education were significant predictors of variance in quality of life (R2 =.35, p =.04). However, mastery, symptom distress, age, and education were found to be significant predictors of variance in quality of life of caregivers (R2 =.40, p =.02). PMID- 12740035 TI - Measuring health-related quality of life in patients with chronic obstructive pulmonary disease in a routine hospital setting: feasibility and perceived value. AB - BACKGROUND: Assessment of health-related quality of life is so far mainly used in specific research settings and not widely accepted in the routine care of patients. Lack of trust in accuracy and reliability and lack of knowledge concerning the questionnaires used, methods, terminology, are just some of the perceived barriers for a more widespread dissemination of these instruments into routine health care. The present study was undertaken in order to test the feasibility of a computerised system for collecting and analysing health-related quality of life in a routine clinical setting and to examine the thoughts and attitudes among physicians concerning the value of these measurements. METHODS: Seventy-four patients with chronic pulmonary lung disease were asked to assess their health-related quality of life with a computerised version of the SF-36 questionnaire before a regular the visit to a physician. The results were immediately available for the physician during the consultation for comparison of information given by the patients and the physician's evaluation of the patients overall health status. A focus group interview with the physicians was performed before and after the implementation of routine measurements of health-related quality of life. RESULTS: The systematic assessment concept worked satisfactorily. All patients approached agreed to participate and completed the assessment on the touch screen computer. A weak correlation was found between patients' self-rated health and pulmonary function and between physicians' evaluation and pulmonary function. The physicians appreciated the SF-36 assessments and the value of the patients' perspective although only a few could pinpoint new clinical decisions based upon this new information. CONCLUSION: Physicians' clinical evaluation and patients' self-rating of health status offer unique and important information that are complementary. PMID- 12740033 TI - 4-aminopyridine decreases progesterone production by porcine granulosa cells. AB - BACKGROUND: Ion channels occur as large families of related genes with cell specific expression patterns. Granulosa cells have been shown to express voltage gated potassium channels from more than one family. The purpose of this study was to determine the effects of 4-aminopyridine (4-AP), an antagonist of KCNA but not KCNQ channels. METHODS: Granulosa cells were isolated from pig follicles and cultured with 4-AP, alone or in combination with FSH, 8-CPT-cAMP, estradiol 17beta, and DIDS. Complimentary experiments determined the effects of 4-AP on the spontaneously established pig granulosa cell line PGC-2. Granulosa cell or PGC-2 function was assessed by radio-immunoassay of media progesterone accumulation. Cell viability was assessed by trypan blue exclusion. Drug-induced changes in cell membrane potential and intracellular potassium concentration were documented by spectrophotometric determination of DiBAC4(3) and PBFI fluorescence, respectively. Expression of proliferating cell nuclear antigen (PCNA) and steroidogenic acute regulatory protein (StAR) was assessed by immunoblotting. Flow cytometry was also used to examine granulosa cell viability and size. RESULTS: 4-AP (2 mM) decreased progesterone accumulation in the media of serum supplemented and serum-free granulosa cultures, but inhibited cell proliferation only under serum-free conditions. 4-AP decreased the expression of StAR, the production of cAMP and the synthesis of estradiol by PGC-2. Addition of either 8 CPT-cAMP or estradiol 17beta to serum-supplemented primary cultures reduced the inhibitory effects of 4-AP. 4-AP treatment was also associated with increased cell size, increased intracellular potassium concentration, and hyperpolarization of resting membrane potential. The drug-induced hyperpolarization of resting membrane potential was prevented either by decreasing extracellular chloride or by adding DIDS to the media. DIDS also prevented 4-AP inhibition of progesterone production. CONCLUSION: 4-AP inhibits basal and FSH-stimulated progesterone production by pig granulosa cells via drug action at multiple interacting steps in the steroidogenic pathway. These inhibitory effects of 4-AP on steroidogenesis may reflect drug-induced changes in intracellular concentrations of K+and Cl- as well as granulosa cell resting membrane potential. PMID- 12740036 TI - Quality of Life measures for dementia. AB - Over the past 10 years, several instruments developed specifically for the assessment of Quality of Life (QOL) in dementia have been introduced. The goal of the current review is to present, compare, and critique existing QOL measures for dementia populations to assist investigators and clinicians in selecting the optimal inventory for their specific needs. Nine measures are reviewed with a focus on conceptualizations of QOL, psychometric data, targeted patient population, and administration and scoring procedures. Critical discussion and comparison of the instruments is presented after the scales are described individually. Differences in definitions of QOL, assessment procedures, and methods that were used to establish the validity of instruments are highlighted. An important direction for future research on QOL scales for dementia is to establish their responsiveness to change over time. It will also be important to identify factors that affect reports of QOL, determine the how perceived QOL affects decisions regarding the care of dementia patients, and evaluate interventions to increase patient QOL. PMID- 12740037 TI - Impact of stress reduction on negative school behavior in adolescents. AB - BACKGROUND: The purpose of this study was to determine the effect of stress reduction via the Transcendental Meditation program on school rule infractions in adolescents. METHODS: Forty-five African American adolescents (ages 15-18 years) with high normal systolic blood pressure were randomly assigned to either Transcendental Meditation (n = 25) or health education control (n = 20) groups. The meditation group engaged in 15-min sessions at home and at school each day for 4 months. The control group was presented 15-min sessions of health education at school each day for 4 months. Primary outcome measures were changes in absenteeism, school rule infractions and suspension days during the four-month pretest period prior to randomization compared with the four-month intervention period. RESULTS: Comparing the pretest and intervention periods, the meditation group exhibited a mean decrease of 6.4 absentee periods compared to an increase of 4.8 in the control group (p <.05). The meditation group exhibited a mean decrease of 0.1 infractions over the four months compared to an increase of 0.3 in the control group (p <.03). There was a mean reduction of 0.3 suspension days due to behavior-related problems in the meditation group compared to an increase of 1.2 in the control group (p <.04). CONCLUSION: These findings demonstrate that the Transcendental Meditation program conducted in the school setting has a beneficial impact upon absenteeism, rule infractions, and suspension rates in African American adolescents. PMID- 12740038 TI - Transthoracic Doppler echocardiography - noninvasive diagnostic window for coronary flow reserve assessment. AB - This review focuses on transthoracic Doppler echocardiography as noninvasive method used to assess coronary flow reserve (CFR) in a wide spectrum of clinical settings. Transthoracic Doppler echocardiography is rapidly gaining appreciation as popular tool to measure CFR both in stenosed and normal epicardial coronary arteries (predominantly in left anterior descending coronary artery). Post stenotic CFR measurement is helpful in: functional assessment of moderate stenosis, detection of significant or critical stenosis, monitoring of restenosis after revascularization. In the absence of stenosis in the epicardial coronary artery, decreased CFR enable to detect impaired microvascular vasodilatation in: reperfused myocardial infarct, arterial hypertension with or without left ventricular hypertrophy, diabetes mellitus, hypercholesterolemia, syndrome X, hypertrophic cardiomyopathy. In these diseases, noninvasive transthoracic Doppler echocardiography allows for serial CFR evaluations to explore the effect of various pharmacological therapies. PMID- 12740040 TI - SLC/CCL21-mediated anti-tumor responses require IFNgamma, MIG/CXCL9 and IP 10/CXCL10. AB - BACKGROUND: SLC/CCL21, normally expressed in high endothelial venules and in T cell zones of spleen and lymph nodes, strongly attracts T cells and dendritic cells (DC). We have previously shown that SLC/CCL21-mediated anti-tumor responses are accompanied by significant induction of IFNgamma and the CXC chemokines, monokine induced by IFNgamma (MIG/CXCL9) and IFNgamma-inducible protein-10 (IP 10/CXCL10). RESULTS: We assessed the importance of IFNgamma, IP-10/CXCL10 and MIG/CXCL9 in SLC/CCL21 therapy. In vivo depletion of IP-10/CXCL10, MIG/CXCL9 or IFNgamma significantly reduced the anti-tumor efficacy of SLC/CCL21. Assessment of cytokine production at the tumor site showed an interdependence of IFNgamma, MIG/CXCL9 and IP-10/CXCL10; neutralization of any one of these cytokines caused a concomitant decrease in all three cytokines. Similarly, neutralization of any one of these cytokines led to a decrease in the frequency of CXCR3+ve T cells and CD11c+ve DC at the tumor site. CONCLUSION: These findings indicate that the full potency of SLC/CCL21-mediated anti-tumor responses require in part the induction of IFNgamma, MIG/CXCL9 and IP-10/CXCL10. PMID- 12740039 TI - Molecular responses to hypoxia in tumor cells. AB - Highly aggressive, rapidly growing tumors are exposed to hypoxia or even anoxia which occurs as a consequence of inadequate blood supply. Both hypoxia and consecutive hypoxia/reoxygenation exert a variety of influences on tumor cell biology. Among these are activation of certain signal transduction pathways and gene regulatory mechanisms, induction of selection processes for gene mutations, tumor cell apoptosis and tumor angiogenesis. Most of these mechanisms contribute to tumor progression. Therefore, tissue hypoxia has been regarded as a central factor for tumor aggressiveness and metastasis. In this review, we summarize the current knowledge about the molecular mechanisms induced by tumor cell hypoxia with a special emphasis on intracellular signal transduction, gene regulation, angiogenesis and apoptosis. Interfering with these pathways might open perspectives for future innovative treatment of highly aggressive metastasizing tumors. PMID- 12740041 TI - The public health benefits of insulation retrofits in existing housing in the United States. AB - BACKGROUND: Methodological limitations make it difficult to quantify the public health benefits of energy efficiency programs. To address this issue, we developed a risk-based model to estimate the health benefits associated with marginal energy usage reductions and applied the model to a hypothetical case study of insulation retrofits in single-family homes in the United States. METHODS: We modeled energy savings with a regression model that extrapolated findings from an energy simulation program. Reductions of fine particulate matter (PM2.5) emissions and particle precursors (SO2 and NOx) were quantified using fuel-specific emission factors and marginal electricity analyses. Estimates of population exposure per unit emissions, varying by location and source type, were extrapolated from past dispersion model runs. Concentration-response functions for morbidity and mortality from PM2.5 were derived from the epidemiological literature, and economic values were assigned to health outcomes based on willingness to pay studies. RESULTS: In total, the insulation retrofits would save 800 TBTU (8 x 10(14) British Thermal Units) per year across 46 million homes, resulting in 3,100 fewer tons of PM2.5, 100,000 fewer tons of NOx, and 190,000 fewer tons of SO2 per year. These emission reductions are associated with outcomes including 240 fewer deaths, 6,500 fewer asthma attacks, and 110,000 fewer restricted activity days per year. At a state level, the health benefits per unit energy savings vary by an order of magnitude, illustrating that multiple factors (including population patterns and energy sources) influence health benefit estimates. The health benefits correspond to 1.3 billion dollars per year in externalities averted, compared with 5.9 billion dollars per year in economic savings. CONCLUSION: In spite of significant uncertainties related to the interpretation of PM2.5 health effects and other dimensions of the model, our analysis demonstrates that a risk-based methodology is viable for national-level energy efficiency programs. PMID- 12740042 TI - A method of drusen measurement based on the geometry of fundus reflectance. AB - BACKGROUND: The hallmarks of age-related macular degeneration, the leading cause of blindness in the developed world, are the subretinal deposits known as drusen. Drusen identification and measurement play a key role in clinical studies of this disease. Current manual methods of drusen measurement are laborious and subjective. Our purpose was to expedite clinical research with an accurate, reliable digital method. METHODS: An interactive semi-automated procedure was developed to level the macular background reflectance for the purpose of morphometric analysis of drusen. 12 color fundus photographs of patients with age related macular degeneration and drusen were analyzed. After digitizing the photographs, the underlying background pattern in the green channel was leveled by an algorithm based on the elliptically concentric geometry of the reflectance in the normal macula: the gray scale values of all structures within defined elliptical boundaries were raised sequentially until a uniform background was obtained. Segmentation of drusen and area measurements in the central and middle subfields (1000 microm and 3000 microm diameters) were performed by uniform thresholds. Two observers using this interactive semi-automated software measured each image digitally. The mean digital measurements were compared to independent stereo fundus gradings by two expert graders (stereo Grader 1 estimated the drusen percentage in each of the 24 regions as falling into one of four standard broad ranges; stereo Grader 2 estimated drusen percentages in 1% to 5% intervals). RESULTS: The mean digital area measurements had a median standard deviation of 1.9%. The mean digital area measurements agreed with stereo Grader 1 in 22/24 cases. The 95% limits of agreement between the mean digital area measurements and the more precise stereo gradings of Grader 2 were -6.4 % to +6.8 % in the central subfield and -6.0 % to +4.5 % in the middle subfield. The mean absolute differences between the digital and stereo gradings 2 were 2.8 +/- 3.4% in the central subfield and 2.2 +/- 2.7% in the middle subfield. CONCLUSIONS: Semi-automated, supervised drusen measurements may be done reproducibly and accurately with adaptations of commercial software. This technique for macular image analysis has potential for use in clinical research. PMID- 12740043 TI - The adoption of Mediterranean diet attenuates the development of acute coronary syndromes in people with the metabolic syndrome. AB - OBJECTIVES: In this work we investigated the effect of the consumption of the Mediterranean diet on coronary risk, in subjects with the metabolic syndrome. METHODS: During 2000-2002, we randomly selected, from all Greek regions, 848 hospitalised patients (695 males, 58 +/- 10 & 153 females, 65 +/- 9 years old) with a first event of acute coronary syndrome and 1078 frequency matched, by sex, age, region controls, without any suspicious for cardiovascular disease. Nutritional habits were evaluated through a validated questionnaire, while the metabolic syndrome was defined according to the NCEP ATP III criteria. Mediterranean diet was defined according to the guidelines of the Division of Nutrition/Epidemiology, of Athens Medical School. RESULTS: Of the 1926 participants, 307 (36.2%) of the patients and 198 (18.4%) of the controls (P < 0.001) met the ATP III criteria. This was related with 2fold adjusted coronary risk (odds ratio = 2.35, 95% 1.87-2.84) in subjects with the metabolic syndrome as compared with the rest of them. No differences were observed concerning the prevalence of the metabolic syndrome and sex of subjects, after adjustment for group of study (P > 0.1). Eighty (26%) of the patients and 70 (35%) of the controls (P < 0.01) with the metabolic syndrome were "closer" to the Mediterranean diet. Multivariate analysis revealed that the adoption of this diet is associated with a 35% (odds ratio = 0.65, 95% 0.44-0.95) reduction of the coronary risk in subjects with the metabolic syndrome, after adjusting for age, sex, educational and financial level and the conventional cardiovascular risk factors. CONCLUSION: Consequently, the adoption of Mediterranean diet seems to attenuate the coronary risk in subjects with the metabolic syndrome. PMID- 12740044 TI - Metabolic networks of microbial systems. AB - In contrast to bioreactors the metabolites within the microbial cells are converted in an impure atmosphere, yet the productivity seems to be well regulated and not affected by changes in operation variables. These features are attributed to integral metabolic network within the microorganism. With the advent of neo-integrative proteomic approaches the understanding of integration of metabolic and protein-protein interaction networks have began. In this article we review the methods employed to determine the protein-protein interaction and their integration to define metabolite networks. We further present a review of current understanding of network properties, and benefit of studying the networks. The predictions using network structure, for example, in silico experiments help illustrate the importance of studying the network properties. The cells are regarded as complex system but their elements unlike complex systems interact selectively and nonlinearly to produce coherent rather than complex behaviors. PMID- 12740045 TI - Cheese whey-induced high-cell-density production of recombinant proteins in Escherichia coli. AB - BACKGROUND: Use of lactose-rich concentrates from dairy processes for the induction of recombinant gene's expression has not received much attention although they are interesting low cost substrates for production of recombinant enzymes. Applicability of dairy waste for induction of recombinant genes in Escherichia coli was studied. Clones expressing Lactobacillus phage muramidase and Lactobacillus alcohol dehydrogenase were used for the experiments. RESULTS: Shake flask cultivations in mineral salt medium showed that cheese whey or deproteinised whey induced gene expression as efficiently as IPTG (isopropyl-beta D-thiogalactopyranoside) or pure lactose. Addition of yeast extract or proteolytically degraded whey proteins did not improve the recombinant protein yield. In contrast, addition of yeast extract to the well-balanced mineral salt medium decreased the product yield. Feeding with glycerol provided sufficient amount of easily assimilable carbon source during the induction period without preventing lactose intake and induction by lactose. High-cell-density fed-batch cultivations showed that product yields comparable to IPTG-induction can be achieved by feeding bacteria with a mixture of glycerol and concentrated whey permeate during the induction. CONCLUSION: Whey and concentrated whey permeate can be applied as an alternative inducer in recombinant high-cell-density fed batch fermentations. The yield of the recombinant product was comparable to fermentations induced by IPTG. In low-cell-density shake flask experiments the yield was higher with whey or whey permeate than with IPTG. PMID- 12740046 TI - Structural basis of regulation and substrate specificity of protein kinase CK2 deduced from the modeling of protein-protein interactions. AB - BACKGROUND: Protein Kinase Casein Kinase 2 (PKCK2) is an ubiquitous Ser/Thr kinase expressed in all eukaryotes. It phosphorylates a number of proteins involved in various cellular processes. PKCK2 holoenzyme is catalytically active tetramer, composed of two homologous or identical and constitutively active catalytic (alpha) and two identical regulatory (beta) subunits. The tetramer cannot phosphorylate some substrates that can be phosphorylated by PKCK2alpha in isolation. The present work explores the structural basis of this feature using computational analysis and modeling. RESULTS: We have initially built a model of PKCK2alpha bound to a substrate peptide with a conformation identical to that of the substrates in the available crystal structures of other kinases complexed with the substrates/ pseudosubstrates. In this model however, the fourth acidic residue in the consensus pattern of the substrate, S/T-X-X-D/E where S/T is the phosphorylation site, did not result in interaction with the active form of PKCK2alpha and is highly solvent exposed. Interaction of the acidic residue is observed if the substrate peptide adopts conformations as seen in beta turn, alpha helix, or 3(10) helices. This type of conformation is observed and accommodated well by PKCK2alpha in calmodulin where the phosphorylation site is at the central helix. PP2A carries sequence patterns for PKCK2alpha phosphorylation. While the possibility of PP2A being phosphorylated by PKCK2 has been raised in the literature we use the model of PP2A to generate a model of PP2A-PKCK2alpha complex. PKCK2beta undergoes phosphorylation by holoenzyme at the N-terminal region, and is accommodated very well in the limited space available at the substrate-binding site of the holoenzyme while the space is insufficient to accommodate the binding of PP2A or calmodulin in the holoenzyme. CONCLUSION: Charge and shape complimentarity seems to play a role in substrate recognition and binding to PKCK2alpha, along with the consensus pattern. The detailed conformation of the substrate peptide binding to PKCK2 differs from the conformation of the substrate/pseudo substrate peptide that is bound to other kinases in the crystal structures reported. The ability of holoenzyme to phosphorylate substrate proteins seems to depend on the accessibility of the P site in limited space available in holoenzyme. PMID- 12740048 TI - Motor effects of short-chain fatty acids and lactate in the gastrointestinal tract. AB - Short-chain fatty acids (SCFA) affect local and remote motility of the gastrointestinal tract by mechanisms that are not completely understood. In the large intestine where they are produced, they inhibit peristaltic activity and may stimulate tonic activity. When present in the terminal ileum as a result of reflux of colon contents, they elicit propulsive contractions. These local motor effects could involve a neuro-hormonal sensory mechanism located in the mucosa of the terminal ileum and proximal colon. Finally, through a humoral pathway probably involving polypeptide YY release, ileal and colonic SCFA modify upper motility by inducing relaxation of the proximal stomach and lower oesophageal sphincter and reducing gastric emptying. One characteristic feature of the SCFA effects is the dose-dependency of the gastrointestinal motor responses. Indeed, the effects occur only below or above a threshold of SCFA concentration in lumen contents. One putative physiological role of the motor effects of SCFA might be to maintain the physico-chemical balance of the lumen environment in the terminal ileum and proximal colon. Another role might be to co-regulate motility of the upper intestine. The clinical relevance of these effects is unclear. However, some recent findings suggest that excessive SCFA concentrations might induce adverse effects on gastrointestinal and colonic motility and sensitivity in certain diseases such as inflammatory bowel disease and gastro-oesophageal reflux disease. PMID- 12740049 TI - Proceedings of the 7th International Vahouny Fibre Symposium. Royal College of Physicians, Edinburgh, United Kingdom, 27-30 May 2002. PMID- 12740047 TI - In situ transduction of target cells on solid surfaces by immobilized viral vectors. AB - BACKGROUND: For both in vitro and in vivo gene transfer applications, recombinant viral vectors have almost always been used free in solution. Some site specificity of the delivery of viral vectors can be achieved by applying a solution containing viral particles specifically to the site of interest. However, such site-specificity is seriously limited since viral vectors can diffuse freely in solution after application. RESULTS: We have developed a novel strategy for in situ transduction of target cells on solid surfaces by viral vectors. In this strategy, adenoviral vectors are attached stably to solid surfaces by using the extremely tight interaction between (strept)avidin and biotin, while maintaining the infectivity of the viral vectors. Target cells are cultured directly on such virus-coated solid surfaces, resulting in the transduction of the cells, in situ, on the solid surface. When compared using an equal number of viral particles present in each well (either immobilized or free), the efficiencies of such in situ transduction on solid surfaces were equivalent to those seen with the adenoviral vectors used free in solution. Since viral particles can be attached at desired locations on solid surfaces in any sizes, shapes, and patterns, the ultimate spatial arrangements of transduced cells on solid surfaces can be predetermined at the time of the preparation of the virus-coated solid surfaces. CONCLUSIONS: We have devised a method of immobilizing adenoviral vectors, tightly and stably, on solid surfaces, while maintaining their ability to infect cells. Such immobilized viral vectors can infect target cells, in situ, on solid surfaces. This strategy should be very useful for the development of a variety of both in vitro and in vivo applications, including the creation of cell-based expression arrays for proteomics and drug discovery and highly site-specific delivery of transgenes for gene therapy and tissue engineering. PMID- 12740050 TI - Dietary fibre analysis. AB - The 'gold standard' method for the measurement of total dietary fibre is that of the Association of Official Analytical Chemists (2000; method 985.29). This procedure has been modified to allow measurement of soluble and insoluble dietary fibre, and buffers employed have been improved. However, the recognition of the fact that non-digestible oligosaccharides and resistant starch also behave physiologically as dietary fibre has necessitated a re-examination of the definition of dietary fibre, and in turn, a re-evaluation of the dietary fibre methods of the Association of Official Analytical Chemists. With this realisation, the American Association of Cereal Chemists appointed a scientific review committee and charged it with the task of reviewing and, if necessary, updating the definition of dietary fibre. It organised various workshops and accepted comments from interested parties worldwide through an interactive website. More recently, the (US) Food and Nutrition Board of the Institute of Health, National Academy of Sciences, under the oversight of the Standing Committee on the Scientific Evaluation of Dietary Reference Intakes, assembled a panel to develop a proposed definition(s) of dietary fibre. Various elements of these definitions were in agreement, but not all. What was clear from both reviews is that there is an immediate need to re-evaluate the methods that are used for dietary fibre measurement and to make appropriate changes where required, and to find new methods to fill gaps. In this presentation, the 'state of the art' in measurement of total dietary fibre and dietary fibre components will be described and discussed, together with suggestions for future research. PMID- 12740051 TI - Dietary fibre intake and clinical indices in the French Supplementation en Vitamines et Mineraux AntioXydants (SU.VI.MAX) adult cohort. AB - The aim of our study was to investigate the relationship between dietary fibre intake and some clinical indices, blood biochemical variables and the incidence of cardiovascular disease and cancers in France, taking advantage of an ongoing cohort, the Supplementation en Vitamines et Mineraux AntioXydants (SU.VI.MAX) intervention study. This preliminary report provides data on dietary fibre intake in this French adult population group of 4080 subjects (2168 men and 1912 women) aged 45-65 years at inclusion. The data obtained for fibre intake indicate that most men and women have low to moderate intakes of total dietary fibre (mean 21.0 and 17.1 g/d respectively), with only 21 % of the men and 7 % of the women having total dietary fibre intakes at the recommended level (i.e. > 25 g/d) and soluble fibre accounting for 19 % of the total dietary fibre intake for both genders. The main food sources of dietary fibre are cereals (30-35 % total), vegetables (20-24 % total) and fruit (19-22 % total). No marked regional differences were observed within France. The highest dietary fibre intakes have been found to be associated with a lower BMI, blood systolic pressure, plasma triacylglycerols and plasma glucose in men and lower BMI in women. Overall, these data support the concept of a beneficial effect of a high dietary fibre intake on cardiovascular disease risk. PMID- 12740052 TI - Dietary fibre in infancy and childhood. AB - There is very little evidence for the effects of dietary fibre in young children and current dietary guidelines are based on assumptions and data extrapolated from studies in adults. The first years of life may be critical for the establishment of a healthy colonic microflora, as well as good eating habits. The lack of clear and well-founded guidelines for the intake of dietary fibre in childhood may hinder both factors. The fears that a high-fibre diet in children < 5 years of age will lead to growth faltering and mineral imbalance are not well supported in the literature, especially for children in the developed world. Indeed, with the rising levels of obesity, fibre intake may be of benefit in reducing energy intake. A low-fibre diet may also be implicated in the aetiology of childhood constipation and appendicitis. The latest proposals for the definition of dietary fibre include oligosaccharides, which may act as prebiotics. There are potential health benefits of including oligosaccharides in the diets of infants and children, but more research is needed to consider the long-term effects. The immature intestine of the infant may also result in a greater amount of starch entering the colon during weaning, and this starch would now be considered dietary fibre under the new definitions. Much new research is needed to allow adequate recommendations for the intake of dietary fibre in childhood based on data collected in appropriate age-groups. PMID- 12740053 TI - Whole grain consumption and weight gain: a review of the epidemiological evidence, potential mechanisms and opportunities for future research. AB - The epidemiological data that directly examine whole grain v. refined grain intake in relation to weight gain are sparse. However, recently reported studies offer insight into the potential role that whole grains may play in body-weight regulation due to the effects that the components of whole grains have on hormonal factors, satiety and satiation. In both clinical trials and observational studies the intake of whole-grain foods was inversely associated with plasma biomarkers of obesity, including insulin, C-peptide and leptin concentrations. Whole-grain foods tend to have low glycaemic index values, resulting in lower postprandial glucose responses and insulin demand. High insulin levels may promote obesity by altering adipose tissue physiology and by enhancing appetite. The fibre content of whole grains may also affect the secretion of gut hormones, independent of glycaemic response, that may act as satiety factors. Future studies may examine whether whole grain intake is directly related to body weight, and whether the associations are primarily driven by components of the grain, including dietary fibre, bran or germ. PMID- 12740054 TI - Colonic diverticula. AB - Diverticulosis is a condition that is associated with ageing. The older the individual the more likely they are to have diverticulosis, but this process is not necessarily inevitable. Diverticula are a herniation through the wall of the sigmoid colon and are likely to be a consequence of a weakness in the colonic wall or prolonged exposure to increased intracolonic pressure consequent on a low dietary fibre intake. The tensile strength of the colon declines with age and becomes least in the distal colon. Adult Africans living in Africa eat a high fibre diet and are free from diverticulosis. Their colons are stronger, wider and thinner than those Scottish adults of the same age. The strength of the colon is in part dependent on collagen fibres and their diameter. The collagen fibrils on the left side of the colon are smaller and more tightly packed than those of the right side of the colon. Ageing and the presence of colonic diverticulosis are associated with smaller more-tightly-packed collagen fibrils. An animal model was established to investigate the effect of a lifelong high- and low-fibre diet on the development of colonic diverticulosis. The findings show that a high-fibre diet from birth, and preferably a maternal high-fibre diet, lessen the risk of diverticulosis with age. PMID- 12740055 TI - On defining dietary fibre. AB - Establishing a definition for dietary fibre has historically been a balance between nutrition knowledge and analytical method capabilities. While the most widely accepted physiologically-based definitions have generally been accurate in defining the dietary fibre in foods, scientists and regulators have tended, in practice, to rely on analytical procedures as the definitional basis in fact. As a result, incongruities between theory and practice have resulted in confusion regarding the components that make up dietary fibre. In November 1998 the president of the American Association of Cereal Chemists (AACC) appointed an expert scientific review committee and charged it with the task of reviewing and, if necessary, updating the definition of dietary fibre. The committee was further charged with assessing the state of analytical methodology and making recommendations relevant to the updated definition. After due deliberation, an updated definition of dietary fibre was delivered to the AACC Board of Directors for consideration and adoption (Anon, 2000; Jones 2000b). The updated definition includes the same food components as the historical working definition used for approximately 30 years (a very important point, considering that the majority of the research of the past 30 years delineating the positive health effects of dietary fibre is based on that working definition). However, the updated definition more clearly delineates the make-up of dietary fibre and its physiological functionality. As a result, relatively few changes will be necessary in analytical methodology. Current methodologies, in particular AACC approved method of analysis 32-05 (Grami, 2000), Association of Official Analytical Chemists' official method of analysis 985.29 (Horwitz, 2000a) or AACC 32-07 (Grami, 2000) Association of Official Analytical Chemists 991.43 (Horwitz, 2000a) will continue to be sufficient and used for most foods. A small number of additional methods will be necessary to quantify the dietary fibre levels in foods containing fibres such as fructans (polymers and oligomers of fructose, inulin), modified dextrins, and/or synthetic dietary fibre analogues. PMID- 12740056 TI - Session: whole cereal grains, fibre and human cancer wholegrain cereals and cancer in Italy. AB - The relationship between frequency of consumption of whole-grain foods and cancer risk has been analysed using data from an updated series of case-control studies conducted in Northern Italy between 1983 and 1997. The overall dataset included the following incident histologically-confirmed neoplasms: oral cavity and pharynx 524, oesophagus 410, stomach 745, colon 955, rectum 625, liver 435, gallbladder 65, pancreas 402, larynx 388, soft tissue sarcomas 217, breast 3412, endometrium 750, ovary 971, prostate 127, bladder 431, kidney 190, thyroid 428, Hodgkin's disease 201, non-Hodgkin's lymphomas 529, multiple myelomas 185. Controls were 10 058 patients admitted to hospital for acute non-neoplastic conditions unrelated to long-term modifications in diet, tobacco or alcohol use. The multivariate odds ratios for the highest category of wholegrain cereal consumption were 0.3-0.5 for upper digestive tract and respiratory neoplasms and colon, 0.6 for rectum and liver, 0.4 for gallbladder, 0.8 for pancreas, 0.2 for soft tissue sarcomas, 0.9 for breast and endometrium, 0.7 for ovary, 0.7 for prostate, 0.4 for bladder and kidney, 1.1 for thyroid and about 0.5 for lymphomas and 0.6 for myelomas. In this population whole-grain food consumption is an indicator of reduced risk of several neoplasms. PMID- 12740057 TI - Can resistant starch and/or aspirin prevent the development of colonic neoplasia? The Concerted Action Polyp Prevention (CAPP) 1 Study. AB - Loss of function of the adenomatous polyposis coli (APC) tumour suppressor gene through truncating mutations or other means is an early event in most colo-rectal cancer (CRC). The APC gene encodes a large multifunctional protein that plays key roles in several cellular processes, including the wnt signalling pathway where an intact APC protein is essential for down regulation of beta-catenin. The APC protein also plays a role in regulation of cell proliferation, differentiation, apoptosis, cell-cell adhesion, cell migration and chromosomal stability during mitosis. Acquisition of a non-functional APC gene can occur by inheritance (in the disease familial adenomatous polyposis (FAP)) or by a sporadic event in a somatic cell. Whilst there is strong epidemiological evidence that variation in diet is a major determinant of variation in CRC incidence, conventional adenoma recurrence trials in sporadic cases of the disease have been relatively unsuccessful in identifying potentially protective food components. Since the genetic basis of CRC in FAP and in sporadic CRC is similar, intervention trials in FAP gene carriers provide an attractive strategy for investigation of potential chemo-preventive agents, since smaller numbers of subjects and shorter time frames are needed. The Concerted Action Polyp Prevention (CAPP) 1 Study is using a 2 x 2 factorial design to test the efficacy of resistant starch (30 g raw potato starch-Hylon VII (1:1, w/w)/d) and aspirin (600 mg/d) in suppressing colo rectal adenoma formation in young subjects with FAP. Biopsies of macroscopically normal rectal mucosa are also being collected for assay of putative biomarkers of CRC risk. PMID- 12740058 TI - Diet and cancer: where are we and where are we going? AB - Many studies have shown that overall dietary patterns, dietary components consumed, or mode of food preparation are all possibly relevant in either reducing or increasing the risk of cancer in animals or man. Yet, dietary intervention studies, stemming from laboratory and epidemiological observational studies have often failed to demonstrate the anticipated protection against cancer. One possible explanation for the discrepancy between the results of various observational and experimental chemo-prevention studies is the lack of control for biological diversity of the participants of these studies. It is suggested that future epidemiological studies provide evidence stratified by status of major metabolic polymorphisms pertinent to the study subject, and that future intervention studies take these differences into account in the design and analysis phases. PMID- 12740059 TI - Dietary fibre and colon cancer: where do we go from here? AB - The relationship between intake of dietary fibre and risk of colon cancer has been studied for 30 years and still the data are inconclusive. There are many possible reasons for this outcome, and they include a failure to consider exposure to dietary fibre separately by source, or colon cancers separately by subsite. These potential confounders have been known for at least 20 years. However, the disease is normally considered by epidemiologists as a single entity. More recently, it has become clear that colon cancer can arise via various histological pathways, and by various genetic pathways. There is no reason at all for assuming that risk factors for these possible pathways are the same. There is a need, therefore, for a more detailed approach to the study of diet and colon cancer, with fibre source and cancer subsite, genetic pathway and histological pathway taken into account. PMID- 12740060 TI - Regulation of short-chain fatty acid production. AB - Short-chain fatty acid (SCFA) formation by intestinal bacteria is regulated by many different host, environmental, dietary and microbiological factors. In broad terms, however, substrate availability, bacterial species composition of the microbiota and intestinal transit time largely determine the amounts and types of SCFA that are produced in healthy individuals. The majority of SCFA in the gut are derived from bacterial breakdown of complex carbohydrates, especially in the proximal bowel, but digestion of proteins and peptides makes an increasing contribution to SCFA production as food residues pass through the bowel. Bacterial hydrogen metabolism also affects the way in which SCFA are made. This outcome can be seen through the effects of inorganic electron acceptors (nitrate, sulfate) on fermentation processes, where they facilitate the formation of more oxidised SCFA such as acetate, at the expense of more reduced fatty acids, such as butyrate. Chemostat studies using pure cultures of saccharolytic gut micro organisms demonstrate that C availability and growth rate strongly affect the outcome of fermentation. For example, acetate and formate are the major bifidobacterial fermentation products formed during growth under C limitation, whereas acetate and lactate are produced when carbohydrate is in excess. Lactate is also used as an electron sink in Clostridium perfringens and, to a lesser extent, in Bacteroides fragilis. In the latter organism acetate and succinate are the major fermentation products when substrate is abundant, whereas succinate is decarboxylated to produce propionate when C and energy sources are limiting. PMID- 12740061 TI - Influences of probiotic bacteria on organic acid production by pig caecal bacteria in vitro. AB - The mechanism of action of probiotics is largely unknown. A potential mechanism should be to increase the production of short-chain fatty acids (SCFA), known modulators of gut functions, by the bacterial ecosystem in the large intestine. The present paper reviews our recent studies in which the capacity of probiotic bacteria to increase the production of SCFA by pig caecal bacteria was investigated using batch-culture and continuous-culture techniques. All four commercial probiotic preparations and three strains of probiotic bacteria dose dependently accelerated the net production of SCFA, succinic acid and lactic acid without changing the acid profile, and slowed the net production of NH4. Effects on organic acid production did not vary among different probiotic species. Neither probiotic preparations nor probiotic bacteria affected the organic acid production from glucose, gastric mucin, starch or lactose, or organic acids produced:added saccharide. Glucose abolished these effects of probiotic preparations. However, the capacity of probiotics to increase SCFA production was not modified by gastric mucin, starch or lactose. These results indicate that probiotic bacteria increase SCFA production by accelerating the breakdown of carbohydrates that are resistant to indigenous bacteria, and suggest that the concept of prebiotics in terms of SCFA production as a measure of probiotic function is arguable. PMID- 12740062 TI - New insight into butyrate metabolism. AB - Butyrate is a C4 acid produced by microbial fermentation of carbohydrates and protein in the large intestine of all animal species. The factor of prime importance for the production rate of butyrate in the lower gut is type and levels of non-digestible carbohydrates entering the large intestine. It was previously believed that 85-90 % of the butyrate produced in the gut was cleared when passing the gut epithelium, but recent studies with catheterised pigs have shown that the concentration of butyrate in the portal vein is strongly influenced by the production rate in the large intestine. Increased gut production of butyrate further raises the circulating level of butyrate. For good reason it is not possible with current technologies to perform direct measurements of the variation in the butyrate concentration in the portal vein of human subjects, but short-chain fatty acid levels in portal blood from sudden death victims, subjects undergoing emergency surgery or planned surgery have indicated a higher gut production and absolute and relative concentration of butyrate in non-fasted as compared with fasted human subjects. However, despite an expected higher gut production of butyrate when feeding a high-fibre rye-bread based diet as compared with a low-fibre wheat-bread-based diet, there was no difference in absolute or relative levels of butyrate in the peripheral blood of human subjects. PMID- 12740063 TI - Production rates and metabolism of short-chain fatty acids in the colon and whole body using stable isotopes. AB - Short-chain fatty acids (SCFA; mainly acetate, propionate and butyrate) are largely produced in non-ruminants during the colonic bacterial fermentation of non-digestible carbohydrates. These intestinal exogenous SCFA pass in part through the splanchnic bed and reach the peripheral bloodstream, mixing with the endogenous circulating SCFA. The whole-body turnover of SCFA is thus composed of an endogenous peripheral turnover and an exogenous production that depends on dietary intake of non-digestible carbohydrates. In the present work methods were developed for determining the SCFA turnover in animals and in human subjects using stable isotopes. The original studies performed to determine endogenous and exogenous metabolism of SCFA in animals and in human subjects are summarised. Using intravenous infusion of 13C-labelled SCFA the whole-body turnover of acetate, propionate and butyrate was assessed in rats in a fasted state. The endogenous turnover of acetate and its oxidation were determined in healthy human subjects in the post-absorptive state, using intravenous infusion of [1 13C]acetate. Intragastric tracer infusions were performed to evaluate the splanchnic first-pass retention of acetate in adults. Finally, an original model was developed in healthy human subjects using intravenous infusion of [1 13C]acetate to determine in vivo the true colonic acetate production after ingestion of a non-digestible disaccharide. These present studies using stable isotopes provide the basis for a novel strategy to evaluate in vivo, in human subjects, the production of SCFA in the large intestine. PMID- 12740064 TI - Molecular analysis of the effect of short-chain fatty acids on intestinal cell proliferation. AB - Short-chain fatty acids (SCFA), particularly butyrate, were shown to regulate cell proliferation in vitro and in vivo. Indeed, butyrate is the major fuel for colonic epithelial cells, and it can influence cell proliferation through the release of growth factors or gastrointestinal peptides such as gastrin, or through modulation of mucosal blood flow. Lastly, SCFA can act directly on genes regulating cell proliferation, and butyrate is the main SCFA to display such an effect. Butyrate inhibits histone deacetylase, which will allow histone hyperacetylation. Such hyperacetylation leads to transcription of several genes, including p21/Cip1. Moreover, it will allow cyclin D3 hyper-expression by inhibiting its degradation. The induction of the cyclin-dependent kinase inhibitory protein p21/Cip1 accounts for cell arrest in the G1 phase of the cell cycle. However, in the absence of p21 other mechanisms are initiated, leading to inhibition of cell proliferation. PMID- 12740065 TI - Anti-cancer effects of butyrate: use of micro-array technology to investigate mechanisms. AB - Epidemiological evidence suggests that a high intake of resistant starch and NSP protects against colo-rectal cancer. The mechanisms underlying this protection are thought to be mediated by the short-chain fatty acid butyrate, which is present in the colonic lumen in millimolar concentrations as a result of bacterial fermentation of carbohydrates that have resisted digestion in the small intestine. In vitro studies have shown that butyrate displays a host of chemo preventative properties including increased apoptosis, reduced proliferation, down regulation of angiogenesis, enhanced immunosurveillance and anti inflammatory effects in colo-rectal cancer cell lines. However, the molecular mechanisms underlying the apparent chemo-preventative actions of butyrate are largely unknown. The evidence supporting the role of butyrate as an anti-cancer agent is reviewed, with particular emphasis on those studies that have attempted to elucidate the mechanism of action of butyrate. Our understanding of the mechanistic action of butyrate and its role in cancer prevention is likely to advance considerably in this post-genomic era with the application of genomic and proteomic technologies. Studies are described that have used gene array and proteomic techniques to investigate the response of colo-rectal cancer cells to butyrate. These pioneering studies illustrate the potential of these technologies to help characterise the molecular responses of the cancer cell to butyrate, and to define the role of butyrate (and other nutrients) in the prevention of colo rectal cancer. PMID- 12740066 TI - Process-induced changes on bioactive compounds in whole grain rye. AB - Manufacturing of healthy wholegrain foods demands knowledge of process-induced changes in macro-, micro- and non-nutrients. The high content of dietary fibre is a challenge in relation to good product texture and sensory quality. The stability and bioavailability of bioactive compounds have a marked influence on the health effects of cereal foods. It was confirmed that sterols, folates, tocopherols and tocotrienols, alkylresorcinols, lignans, phenolic acids and total phenolics are concentrated in the bran layers of the rye grain, and are only present at low levels in the flour endosperm. The levels of folate and easily extractable phenolic compounds increase in germination and sourdough baking, but there are negligible changes in the levels of sterols, lignans and alk(en)ylresorcinols. The levels of tocopherols and tocotrienols are reduced during the sourdough fermentation. In conclusion, many of the bioactive compounds in wholegrain rye are stable during food processing, and their levels can even be increased with suitable processing. PMID- 12740067 TI - Why whole grains are protective: biological mechanisms. AB - Epidemiological studies find that whole-grain intake is protective against cancer, cardiovascular disease, diabetes and obesity. Potential mechanisms for this protection are diverse since whole grains are rich in nutrients and phytochemicals. First, whole grains are concentrated sources of dietary fibre, resistant starch and oligosaccharides, carbohydrates that escape digestion in the small intestine and are fermented in the gut, producing short-chain fatty acids (SCFA). SCFA lower colonic pH, serve as an energy source for the colonocytes and may alter blood lipids. These improvements in the gut environment may provide immune protection beyond the gut. Second, whole grains are rich in antioxidants, including trace minerals and phenolic compounds, and these compounds have been linked to disease prevention. Additionally, whole grains mediate insulin and glucose responses. Although lower glycaemic load and glycaemic index have been linked to diabetes and obesity, risk of cancers such as colon and breast cancer have also been linked to high intake of readily-available carbohydrate. Finally, whole grains contain many other compounds that may protect against chronic disease. These compounds include phytate, phyto-oestrogens such as lignan, plant stanols and sterols, and vitamins and minerals. As a consequence of the traditional models of conducting nutrition studies on isolated nutrients, few studies exist on the biological effects of increased whole-grain intake. The few whole-grain feeding studies that are available show improvements in biomarkers with whole-grain consumption, such as weight loss, blood lipid improvement and antioxidant protection. PMID- 12740068 TI - Whole grains protect against atherosclerotic cardiovascular disease. AB - Atherosclerotic cardiovascular disease (ASCVD) is the most common cause of death in most Western countries. Nutrition factors contribute importantly to this high risk for ASCVD. Favourable alterations in diet can reduce six of the nine major risk factors for ASCVD, i.e. high serum LDL-cholesterol levels, high fasting serum triacylglycerol levels, low HDL-cholesterol levels, hypertension, diabetes and obesity. Wholegrain foods may be one the healthiest choices individuals can make to lower the risk for ASCVD. Epidemiological studies indicate that individuals with higher levels (in the highest quintile) of whole-grain intake have a 29 % lower risk for ASCVD than individuals with lower levels (lowest quintile) of whole-grain intake. It is of interest that neither the highest levels of cereal fibre nor the highest levels of refined cereals provide appreciable protection against ASCVD. Generous intake of whole grains also provides protection from development of diabetes and obesity. Diets rich in wholegrain foods tend to decrease serum LDL-cholesterol and triacylglycerol levels as well as blood pressure while increasing serum HDL-cholesterol levels. Whole-grain intake may also favourably alter antioxidant status, serum homocysteine levels, vascular reactivity and the inflammatory state. Whole-grain components that appear to make major contributions to these protective effects are: dietary fibre; vitamins; minerals; antioxidants; phytosterols; other phytochemicals. Three servings of whole grains daily are recommended to provide these health benefits. PMID- 12740069 TI - Epidemiological support for the protection of whole grains against diabetes. AB - The epidemic of type 2 diabetes among children, adolescents and adults is increasing along with the increasing prevalence of overweight and obesity. Overweight is the most powerful modifiable risk factor for type 2 diabetes. Intake of wholegrain foods may reduce diabetes risk. Three prospective studies in 160000 men and women examined the relationship of whole-grain or cereal-fibre intake with the risk of type 2 diabetes. Each study used a mailed Willett food frequency questionnaire and similar methods of quantifying wholegrain foods and cereal fibre. The self-reported incident diabetes outcome was more reliably determined in the two studies of health-care professionals than in the study of Iowa women. Risk for incident type 2 diabetes was 21-27% lower for those in the highest quintile of whole-grain intake, and 30-36% lower in the highest quintile of cereal-fibre intake, each compared with the lowest quintile. Risk reduction persisted after adjustment for the healthier lifestyle found among habitual whole grain consumers. Observations in non-diabetic individuals support an inverse relationship between whole-grain consumption and fasting insulin levels. In feeding studies in non-diabetic individuals insulin resistance was reduced using whole grains or diets rich in whole grains. Glucose control improved with diets rich in whole grains in feeding studies of subjects with type 2 diabetes. There is accumulating evidence to support the hypothesis that whole-grain consumption is associated with a reduced risk of incident type 2 diabetes; it may also improve glucose control in diabetic individuals. PMID- 12740070 TI - What are the responsibilities of Public Health Nutritionists in the context of war? PMID- 12740071 TI - Out of the box. PMID- 12740072 TI - National immunisation days and vitamin A distribution in Mali: has the vitamin A status of pre-school children improved? AB - OBJECTIVES: The impact on vitamin A status of the distribution of vitamin A during national immunisation days (NIDs) has not been well established despite strong promotion by international agencies and donors. Using a pre-post design, the change in prevalence of vitamin A deficiency was examined in pre-school children in Mali. DESIGN: Two cross-sectional surveys were conducted in Mopti region, the first in March 1997 before this strategy was adopted and the second in March 1999, four-and-a-half months after a mass distribution of vitamin A during NIDs. SUBJECTS AND SETTING: We compared the vitamin A status of children aged 12 to 66 months targeted in 1999 by NIDs with the status of children in the same age group in 1997. Infectious events of the previous two weeks were concurrently recorded. Within the 1999 sample, the status of recipient and non recipient children was also compared. RESULTS: In 1997, the prevalence of xerophthalmia (defined by the presence of night blindness and/or Bitot spots) was 6.9% (95% confidence interval (CI) 5.1-9.2) and the modified retinol dose response (MRDR) test proved abnormal in 77.8% of 12-66-month-old children (95% CI 68.27-85.17). In 1999 this picture had improved significantly, both for xerophthalmia prevalence, 3.3% (95% CI 2.1-5.2), and abnormal MRDR test response, 63.1% (95% CI 54.25-71.23). The infectious morbidity rates between 1997 and 1999 tended to decrease. No significant improvement was found among children older than those targeted by NIDs. In 1999, children who received vitamin A had a lower risk for xerophthalmia (3.0% for recipients vs. 8.7% for non-recipients) and experienced fewer infectious events. CONCLUSIONS: The clinical and biological vitamin A status of pre-school children improved between 1997 and 1999. Mass distribution of vitamin A appears to reduce the occurrence of xerophthalmia and would seem to be associated with a decrease in other related illnesses. Vitamin A supplementation during NIDs should be given a high priority when vitamin A deficiency remains a public health problem. PMID- 12740073 TI - Human milk fatty acids and growth of infants in Brazzaville (The Congo) and Ouagadougou (Burkina Faso). AB - OBJECTIVE: To estimate the role of human milk n-6 and n-3 polyunsaturated fatty acids (PUFA) in term infant growth in two African urban populations. DESIGN: Observational study. Weight gains at 5 months of age and dietary habits were compared between Congolese infants (n=102) and Burkinabe infants (n=101). Socio economic status and anthropometry of the mothers were also recorded. SETTING: One suburban district in Brazzaville (capital of The Congo) and one in Ouagadougou (capital of Burkina Faso). SUBJECTS: Two random samples of nursing mothers and their 5-month-old infants. RESULTS: All infants were born at term and there was no difference in birth weights. At 5 months of age, infants in Ouagadougou were thinner but not shorter than their counterparts in Brazzaville (average weight gain (standard deviation): 614 (168) g month-1 vs. 720 (176) g month-1; P= 0.0001). Drastic differences were found in infant diets with regard to extra fluid intake and n-6 and n-3 PUFA concentrations in breast milk. In Ouagadougou, all infants were given fluids other than milk from birth. Breast milk had highly unbalanced 18:2n-6/18:3n-3 and n-6/n-3 long-chain PUFA ratios (53:1 and 5:1, respectively). In Brazzaville, half of the infants received fluids other than milk, and breast milk showed balanced 18:2n-6/18:3n-3 and n-6/n-3 long-chain PUFA ratios (12:1 and 1:1, respectively). A non-linear relationship between 18:2n 6/18:3n-3 ratio and growth was established in Brazzaville (P= 0.0027). The 18:2n 6/18:3n-3 ratio adjusted with covariates had an even more significant effect on weight gain (P= 0.0011). Applying the same model in Ouagadougou did not show such a relation. CONCLUSION: : Data strongly suggest that a balanced ratio of 18:2n 6/18:3n-3 (between 5:1 and 15:1) in breast milk leads to higher weight gain of infants during the first 5 months of life. PMID- 12740074 TI - Does the consumption of amylase-containing gruels impact on the energy intake and growth of Congolese infants? AB - OBJECTIVE: To assess the effect of the incorporation of amylase in maize-based flours prepared as gruels on the energy intake and growth of Congolese infants. DESIGN: A randomised controlled trial. At 18 weeks of age, infants were randomised into either an intervention group, where they were provided with a maize/soya-based flour that contained amylase, or a control group, where they were provided with a similar flour that did not contain amylase. SETTING: Urban borough of Poto-Poto in Brazzaville, The Congo. SUBJECTS: Eighty infants (40 in each group) were randomised into intervention and control groups. Three infants in the intervention group and two controls subsequently dropped out. RESULTS: At 24 weeks, the addition of amylase resulted in a significant increase in energy intake (in kJ kg-1 day-1) from gruels without affecting breast milk consumption. In contrast, total energy intake (in kJ kg-1 day-1) did not differ significantly between groups (P= 0.08). After adjustment for morbidity and previous growth, infants in the intervention group showed better growth in length during the trial (+0.22 cm month-1; P= 0.04), especially between 24 and 31 weeks of age (+0.51 cm month-1; P< 0.01). There were no differences in weight velocity between groups. CONCLUSIONS: The findings of this study suggest that the consumption of amylase treated gruels allows an increase in energy intake from these gruels without affecting breast milk consumption but has no impact on total daily energy intake. However, if started after 6 months, it could be effective in preventing faltering of infant linear growth. PMID- 12740075 TI - EPIC-Oxford: lifestyle characteristics and nutrient intakes in a cohort of 33 883 meat-eaters and 31 546 non meat-eaters in the UK. AB - OBJECTIVE: To describe the lifestyle characteristics and nutrient intakes of the Oxford cohort of the European Prospective Investigation into Cancer and Nutrition (EPIC). DESIGN: Cohort of men and women recruited through general practices or by post to include a high proportion of non meat-eaters. Dietary, anthropometric and lifestyle data were collected at baseline and four diet groups were defined. SETTING: United Kingdom. PARTICIPANTS: In total, 65 429 men and women aged 20 to 97 years, comprising 33 883 meat-eaters, 10 110 fish-eaters, 18 840 lacto-ovo vegetarians and 2596 vegans. RESULTS: Nutrient intakes and lifestyle factors differed across the diet groups, with striking differences between meat-eaters and vegans, and fish-eaters and vegetarians usually having intermediate values. Mean fat intake in each diet group was below the UK dietary reference value of 33% of total energy intake. The mean intake of saturated fatty acids in vegans was approximately 5% of energy, less than half the mean intake among meat-eaters (10-11%). Vegans had the highest intakes of fibre, vitamin B1, folate, vitamin C, vitamin E, magnesium and iron, and the lowest intakes of retinol, vitamin B12, vitamin D, calcium and zinc. CONCLUSIONS: The EPIC-Oxford cohort includes 31 546 non meat-eaters and is one of the largest studies of vegetarians in the world. The average nutrient intakes in the whole cohort are close to those currently recommended for good health. Comparisons of the diet groups show wide ranges in the intakes of major nutrients such as saturated fat and dietary fibre. Such variation should increase the ability of the study to detect associations of diet with major cancers and causes of death. PMID- 12740076 TI - Patterns and demographic predictors of 5-year weight change in a multi-ethnic cohort of men and women in Australia. AB - OBJECTIVE: This study investigated 5-year trends in body weight, overweight and obesity and their association with sociodemographic variables in a large, multi ethnic community sample of Australian adults. DESIGN: This prospective population study used baseline and 5-year follow-up data from participants in the Melbourne Collaborative Cohort Study (MCCS). SETTING: Population study in Melbourne, Australia. SUBJECTS: In total, 12 125 men and 17 674 women aged 35-69 years at baseline. RESULTS: Mean 5-year weight change in this sample was +1.58 (standard deviation (SD) 4.82) kg for men and +2.42 (SD 5.17) kg for women. Younger (35-44 years) men and, in particular, women gained more weight than older adults and were at highest risk of major weight gain (> or =5 kg) and becoming overweight. Risk of major weight gain and associations between demographic variables and weight change did not vary greatly by ethnicity. Education level showed complex associations with weight outcomes that differed by sex and ethnicity. Multivariate analyses showed that, among men, higher initial body weight was associated with decreased likelihood of major weight gain, whereas among women, those initially overweight or obese were about 20% more likely to experience major weight gain than underweight or healthy weight women. CONCLUSIONS: Findings of widespread weight gain across this entire population sample, and particularly among younger women and women who were already overweight, are a cause for alarm. The prevention of weight gain and obesity across the entire population should be an urgent public health priority. Young-to-mid adulthood appears to be a critical time to intervene to prevent future weight gain. PMID- 12740077 TI - Risk analysis applied to food fortification. AB - OBJECTIVE: To describe how a risk analysis can be applied to food fortification, with emphasis on voluntary fortification and intake levels that might exceed usual dietary levels. DESIGN: Use of the risk analysis model as a frame to classify nutrients according to the risk of exceeding upper safe intake levels. Furthermore, to apply the model when discussing possible consequences of liberal fortification practices on eating behaviour and disease patterns. SETTING: The discussion on food fortification presently going on internationally. RESULTS: Micronutrients can be classified according to their safety margin, i.e. the size of the interval between the recommended intake and the upper safe level of intake. We suggest that nutrients with a small safety margin, i.e. for which the upper safe level is less than five times the recommended intake, be placed in a category A and should be handled with care (retinol, vitamin D, niacin, folate and all minerals). Category B comprises nutrients with an intermediate safety margin (vitamins E, B6, B12 and C), while nutrients that according to present knowledge are harmless even at 100 times the recommendation (vitamin K, thiamin, riboflavin, pantothenic acid and biotin) are categorised as C. DISCUSSION: The risk analysis model is a useful tool when assessing the risk of both too low and excess intakes of single micronutrients, but can also be applied to analyse the consequences of fortification practices on eating behaviour and disease patterns. Liberal fortification regulations may, for example, distort the conception of what is healthy food, and drive consumption towards a more unhealthy diet, contributing to the plague of overweight and concomitant increased risk of degenerative diseases. CONCLUSION: The impact of fortification practices on the total eating pattern of a population should become an integrated part of the discussions and regulations connected to the issue. PMID- 12740078 TI - Sampling procedure, participation rates and representativeness in the Swedish part of the European Youth Heart Study (EYHS). AB - OBJECTIVE: The European Youth Heart Study (EYHS) is a cross-sectional, school based population study on risk factors for future cardiovascular disease in children, with an overall participation rate in Sweden of about 50%. To study the representativeness of the participants in the Swedish part of EYHS, a comprehensive non-participant follow-up study was carried out. DESIGN: A structured multilevel analysis model was developed, addressing each level in the sampling procedure. The income, educational and occupational categories of the geographical regions of the study (level I), school catchment areas (level II) and parents (level III) were compared with official data. Participating and non participating pupils (level IV) were compared through a questionnaire. SETTING: Thirty-seven state schools in two regions of Central Sweden (Orebro and southern Stockholm) were visited during the school year 1998/1999. SUBJECTS: Boys and girls aged 9 and 15 years were randomly sampled through a multiphase sampling procedure. RESULTS: Data for socio-economic status for levels I and II corresponded well to national and regional official data. At level III, non manually working parents were slightly over-represented among parents of participating children. At level IV, non-participating subjects corresponded in most respects to participants with a few exceptions--mainly more interest in physical exercise among participants. CONCLUSIONS: Based on the knowledge from the non-participant study, we do not foresee problems regarding interpretation of the outcomes in the EYHS, despite the low participation rate. PMID- 12740079 TI - Addressing overreporting on the International Physical Activity Questionnaire (IPAQ) telephone survey with a population sample. AB - OBJECTIVE: To examine a possible problem of overreporting and to describe the degree of error with the International Physical Activity Questionnaire (IPAQ) short telephone protocol. DESIGN: Cross-sectional study, using two different physical activity (PA) self-report protocols. SETTING: Telephone interviews about PA in Belgium. SUBJECTS: Fifty adults who had previously been interviewed with IPAQ in a national survey. RESULTS: Seventy-five per cent reported less PA with the modified procedure than with the IPAQ. Twenty-three of the 50 individuals were found to have reported some amounts of PA with the IPAQ (either walking, or vigorous or moderate PA) when they should have reported none. In total, based on their revised reports of PA, 50% fewer persons met PA recommendations than was the case with IPAQ. The overreporting could not be related to types of error prone individuals. CONCLUSIONS: Overreporting of PA in population samples is a serious problem that could be reduced by implementing procedure changes without changing the IPAQ items themselves. PMID- 12740080 TI - Comparison of body mass index values proposed by Cole et al. (2000) and Must et al. (1991) for identifying obese children with weight-for-height index recommended by the World Health Organization. AB - OBJECTIVES: To calculate the sensitivity, specificity and agreement of body mass index (BMI) values proposed by Cole et al. (Br. Med. J. 2000; 320: 1) and Must et al. (Am. J. Clin. Nutr. 1991; 53: 839 & 54: 773) with weight-for-height index in the nutritional evaluation of children. DESIGN: Criterion standards for diagnostic tests. SETTING: : North-east and south-east Brazil. SUBJECTS: Two thousand nine hundred and twenty children studied in Life Pattern Research performed by the Brazilian Institute of Geography and Statistics in 1997. Main outcome measures are the sensitivity, specificity and agreement of BMI values proposed by Must et al. (1991) and Cole et al. (2000). RESULTS: Sensitivity of values proposed by both authors was around 90%. Specificity was almost 100% considering weight-for-height index as the gold standard. The agreement of both values with weight-for-height index, based on kappa results, was good and in pre school children it was excellent. CONCLUSIONS: Values proposed by Cole et al. (2000) and Must et al. (1991) should be used carefully to screen obesity in childhood but can be used to "diagnose" overweight children with a very low chance of having false-positive results. Although the values proposed by both authors performed similarly, use of Cole et al.'s values should be encouraged. The latter cover children from 2 to 6 years old; their values are presented for six-month age intervals; they are based on a larger sample from six different countries; and they are related to the definition of adult obesity. PMID- 12740081 TI - Statistical approaches for assessing the relative validity of a food-frequency questionnaire: use of correlation coefficients and the kappa statistic. AB - OBJECTIVE: To compare different statistical methods for assessing the relative validity of a self-administered, 150-item, semi-quantitative food-frequency questionnaire (FFQ) with 4-day weighed diet records (WR). DESIGN: Subjects completed the Scottish Collaborative Group FFQ and carried out a 4-day WR. Relative agreement between the FFQ and WR for energy-adjusted nutrient intakes was assessed by Pearson and Spearman rank correlation coefficients, the percentages of subjects classified into the same and opposite thirds of intake, and Cohen's weighted kappa. SUBJECTS: Forty-one men, mean age 36 (range 21-56) years, and 40 women, mean age 33 (range 19-58) years, recruited from different locations in Aberdeen, Scotland. RESULTS: Spearman correlation coefficients tended to be lower than Pearson correlation coefficients, and were above 0.5 for 10 of the 27 nutrients in men and 17 of the 27 nutrients in women. For nutrients with Spearman correlation coefficients above 0.5, the percentage of subjects correctly classified into thirds ranged from 39 to 78%, and weighted kappa values ranged from 0.23 to 0.66. CONCLUSIONS: Both Spearman correlation coefficients and weighted kappa values are useful in assessing the relative validity of estimates of nutrient intake by FFQs. Spearman correlation coefficients above 0.5, more than 50% of subjects correctly classified and less than 10% of subjects grossly misclassified into thirds, and weighted kappa values above 0.4 are recommended for nutrients of interest in epidemiological studies. PMID- 12740082 TI - Tools and techniques for craniofacial tissue engineering. AB - Craniofacial surgery is an important conduit for tissue-engineering applications. As interdisciplinary collaborations improve, we can expect to see remarkable progress in de novo tissue synthesis, replacement, and repair. Ultimately, we may one day find that gene-modified cell-based tissue-engineering strategies will succeed today's reconstructive strategies. In this review, we highlight the major gene- and cell-based preclinical tools and techniques that are currently being developed to solve common craniofacial problems. PMID- 12740083 TI - Spatial confinement of neurite regrowth from dorsal root ganglia within nonporous microconduits. AB - Tissue engineering is founded on the concept of controlling the behavior of individual cells to stimulate tissue formation. This control is achieved by mimicking signals that manage natural tissue development or repair. These interdependent signals include cytokine delivery, extracellular matrix interactions, and cell-cell communication. Here, we report on the effect of spatial guidance as a signal for nerve tissue regeneration, using a simple in vitro model. We observe the acceleration of neurite extension from rat dorsal root ganglia within micron-scale tubes. Within these hydrogel-filled conduits, neurites were observed to extend more rapidly than when cultured within the hydrogel alone. The spatial cue also induced a change in tissue architecture, with the cabling of cells within the microconduit. The acceleration of neurite extension was found to be independent of conduit diameter within the range of 200 to 635 microm. Finally, our in vitro model enabled quantification of the effect of combining spatial control and localized nerve growth factor delivery. PMID- 12740084 TI - Addition of fibronectin to alginate matrix improves peripheral nerve regeneration in tissue-engineered conduits. AB - Schwann cell (SC) transplantation has been proposed to encourage peripheral nerve regeneration, but an optimal SC-carrying matrix would be needed. The aim of this study was to characterize how the addition of fibronectin to alginate would affect the outcome of nerve regeneration promoted by Schwann cells embedded in this matrix. Genetically labeled rat SCs were obtained by lacZ gene transduction. SCs were suspended in alginate hydrogel matrix with/without addition of liquid fibronectin, and their viability and growth in the different types of matrices were assessed in vitro by AlamarBlue assay. In vivo assessment of SC transplantation in the matrix was carried out with poly-3-hydroxybutyrate (PHB) conduits to bridge a sciatic nerve gap. The grafted conduits were harvested at 2, 3, and 6 weeks and assessed for the presence of labeled SCs in relation to regrowing axons. The amount and rate of axonal regeneration were assessed by quantitative immunohistochemistry. Addition of fibronectin to alginate hydrogel improved SC viability and growth profile in vitro. X-Gal staining confirmed that SCs transplanted in PHB conduits were viable throughout the time course, and that the labeled SCs were clearly associated with regenerating axons. The regeneration rate was enhanced when liquid fibronectin was added to the alginate matrix. Furthermore, the presence of SCs also enhanced regeneration and there was an additive effect when both SCs and fibronectin were combined with alginate. In conclusion, the addition of fibronectin to alginate hydrogel matrix contributed to improve nerve regeneration, supporting SC viability and augmenting their effect on axonal growth when transplanted in a bioengineered nerve conduit. PMID- 12740085 TI - Nerve guide material made from fibronectin: assessment of in vitro properties. AB - We have previously used orientated mats of fibronectin as conduits to repair short gaps in peripheral nerves. Here we describe the in vitro properties of a new material in the form of large cables produced from a fibronectin-enriched solution with potential as a conduit for longer nerve defects. Large cables of fibronectin were made up to 14 cm long x 1.5 cm in diameter. When freeze dried, scanning electron microscopy revealed a predominant fiber orientation. Dried cables hydrated rapidly to 1.6 and 4.8 times their original length and diameter, respectively. Once hydrated these cables had pores that ranged from 10 to 100 microm through which Schwann cells and fibroblasts were able to grow in vitro and align with the axis of the fibrils by contact guidance. Furthermore, the porosity of the cable was enhanced by the natural dissolution of protein over a 3-week duration in culture with cells, such that 50- to 200-microm pores were observed. This study suggests that large fibronectin cables are a suitable alternative to the original fibronectin mats to guide components of the peripheral nerves and so to act as conduits with potential use in guiding regeneration across long nerve defects. PMID- 12740086 TI - Optimization of Schwann cell adhesion in response to shear stress in an in vitro model for peripheral nerve tissue engineering. AB - The design of nerve guidance channels (NGCs) is evolving to produce a favorable environment for neural regeneration. We created an in vitro model to evaluate the interactions between three centrally important components of this altered host environment: (1). Schwann cells, (2). substrate, and (3). sustained mechanical stimulus in the form of shear stress with laminar fluid flow. Preconfluent Schwann cells were plated on slides coated either with laminin, poly-D-lysine, type IV collagen, or fibronectin. These slides were placed into custom-designed, parallel-plate, flow chambers and were administered laminar fluid flow at a rate of 15 mL/min for 2 h. Schwann cell adhesion assays demonstrated that laminin (mean, 86.1%; SEM, 4.47%) and fibronectin (mean, 81.7%; SEM, 3.24%) were statistically superior to collagen type IV (mean, 57.7%; SEM, 3.96%) and poly-D lysine (mean, 58.0%; SEM, 4.97%) (p < 0.001). Fibronectin (mean, 12.20%; SEM, 0.374%) induced statistically greater Schwann cell proliferation than did laminin (mean, 8.14%; SEM, 0.682%) (p < 0.001). Therefore, we recommend that fibronectin should be used as an important component of NGCs with further in vivo studies. As mechanical stress is an integral part of the host environment, our study is the first to incorporate this factor into an in vitro model for peripheral nerve tissue engineering. PMID- 12740087 TI - Crosslinking characteristics of and cell adhesion to an injectable poly(propylene fumarate-co-ethylene glycol) hydrogel using a water-soluble crosslinking system. AB - The crosslinking characteristics of an injectable poly(propylene fumarate-co ethylene glycol) [P(PF-co-EG)]-based hydrogel were investigated. A water-soluble crosslinking system was used, consisting of poly(ethylene glycol) diacrylate (PEG DA), ammonium persulfate (APS), and ascorbic acid (AA). The effects of PEG block length of the P(PF-co-EG), APS concentration, AA concentration, and PEG-DA concentration on equilibrium water content, sol fraction, onset of gelation, mechanical properties, and endothelial cell adhesion were studied. The equilibrium water content of the hydrogels ranged from 57.1 +/- 0.3 to 79.7 +/- 0.2% whereas the sol fraction ranged from 2.5 +/- 0.0 to 3.33 +/- 5.4%. The onset of gelation times varied from 1.1 +/- 0.1 to 4.3 +/- 0.2 min. For all hydrogel formulations, the tensile strength fell between 61.7 +/- 18.2 and 401.3 +/- 67.5 kPa and tensile moduli ranged from 0.4 +/- 0.0 to 3.3 +/- 0.3 MPa. Endothelial cells attached to the hydrogels in a range of 3.9 +/- 1.4 to 31.1 +/- 14.1% of cells seeded. These findings suggest that injectable poly(propylene fumarate-co ethylene glycol) hydrogel formulations in conjunction with a novel water-soluble crosslinking system may be useful for in situ crosslinkable tissue-engineering applications. PMID- 12740088 TI - Microfluidic patterning of cells in extracellular matrix biopolymers: effects of channel size, cell type, and matrix composition on pattern integrity. AB - The organization of cells within an extracellular matrix is critical to promote appropriate cellular interactions and tissue function in vivo. The ability to design and create biologically relevant cellular arrangements via microfluidic patterning on surfaces provides new capabilities for tissue engineering and biomimetics. The purpose of this article is to describe techniques using microfluidic patterning of three-dimensional biopolymer matrices to improve cellular pattern integrity and to provide microscale control over cellular microenvironments. Results demonstrated that the incorporation of extracellular matrix biopolymers in cell microfluidic patterning results in a more stable pattern of adherent human endothelial cells than patterning without matrix components after several days in vitro. This may be important for carrying out long-term biological experiments and tissue engineering in vitro. Moreover, chemical components in the patterned biopolymer matrices, such as collagen, chitosan, and fibronectin, influenced the ability of the matrices to control cell migration and pattern stability over time. Thus, microfluidic patterning of cells in extracellular matrix biopolymers was shown to be useful in patterning multiple cell types in well-defined three-dimensional geometries. PMID- 12740089 TI - Monitoring local cell viability in engineered tissues: a fast, quantitative, and nondestructive approach. AB - Assessment of cell viability is a key issue in monitoring in vitro engineered tissue constructs. In this study we describe a fully automated, quantitative, and nondestructive approach, which is particularly suitable for tissue engineering. The approach offers several advantages above existing methods. Living and dead cell numbers can be separately determined for both isolated cells and cells that form networks during tissue formation. Moreover, viability can be locally monitored in time throughout the three-dimensional tissue. The viability assay is based on a dual fluorescent staining technique using CellTracker Green (CTG) for detection of living cells and propidium iodide (PI) for dead cells. CTG and PI images are created with a confocal laser scanning microscope. To determine the number of living cells, CTG fluorescence intensity is determined from the CTG image. Thereby, novel image-processing techniques have been developed, normalizing for various undesired influences that alter measurements of absolute CTG fluorescence intensities. Dead cell numbers are determined from the PI image, using an improved computerized counting method. The approach was first evaluated on C2C12 monolayers, of which images were taken directly after probe addition and 24 h later. Results show that at both times, computed living and dead cell numbers highly correlate with manually counted cell numbers (r > 0.996). Next, the approach was applied for monitoring viability in three-dimensional engineered skeletal muscle tissue constructs, which were subjected to unfavorable environmental conditions. This example illustrated that local viability can be quantitatively, nondestructively, and locally monitored in three-dimensional tissue constructs, making it a promising tool in the field of tissue engineering. PMID- 12740090 TI - Analytical model for predicting mechanotransduction effects in engineered cardiac tissue. AB - Mechanochemical and mechanoelectrical signaling is imperative for cardiac organogenesis and underlies pathophysiological events. New techniques for engineering cardiac tissue allow unprecedented means of modeling these phenomena in vitro. However, experimental design is often hampered by a lack of models that can be adapted to the ideal conditions these methods allow. To address these deficiencies, we developed a mathematical model to calculate the distribution of stress and strain in fibrous cardiac tissue. The fluid-fiber-collagen model characterizes the mechanical behavior of cardiac tissue and is solved analytically for the distributions of stress and strain along the myocardial fibers. An example application of the model is presented: modeling the distribution of strains in the vicinity of an ischemic region. The ischemic region is stretched during systole, as has been shown in previous one-dimensional models. Our model predicts a complex distribution of stretch in the border zone surrounding the ischemic region and in nonischemic regions surrounding the border zone. These strain patterns may predict patterns of mechanochemical coupling that results in localized fibrosis, altered gene expression, or the mechanoelectrical signaling events that potentiate cardiac arrhythmias. PMID- 12740091 TI - Dynamic rotational seeding and cell culture system for vascular tube formation. AB - Optimization of cell seeding and culturing is an important step for the successful tissue engineering of vascular conduits. We evaluated the effectiveness of using a hybridization oven for rotational seeding and culturing of ovine vascular myofibroblasts onto biodegradable polymer scaffolds suitable for replacement of small- and large-diameter blood vessels. Large tubes (12 mm internal diameter and 60 mm length, n = 4) and small tubes (5 mm internal diameter and 20 mm length, n = 4) were made from a combination of polyglycolic acid/poly-4-hydroxybutyrate and coated with collagen solution. Tubes were then placed in culture vessels containing a vascular myofibroblast suspension (10(6) cells/cm(2)) and rotated at 5 rpm in a hybridization oven at 37 degrees C. Light and scanning electron microscopy analyses were performed after 5, 7, and 10 days. Myofibroblasts had formed confluent layers over the outer and inner surfaces of both large and small tubular scaffolds by day 5. Cells had aligned in the direction of flow by day 7. Multiple spindle-shaped cells were observed infiltrating the polymer mesh. Cell density increased between day 5 and day 10. All conduits maintained their tubular shape throughout the experiment. We conclude that dynamic rotational seeding and culturing in a hybridization oven is an easy, effective, and reliable method to deliver and culture vascular myofibroblasts onto tubular polymer scaffolds. PMID- 12740092 TI - Engineering of vaginal tissue in vivo. AB - Congenital vaginal anomalies and cloacal malformations may require extensive surgical reconstruction. Surgical challenges are often encountered because of the limited amounts of native tissue available. We investigated the feasibility of using vaginal epithelial and smooth muscle cells for the engineering of vaginal tissues in vivo. Vaginal epithelial and smooth muscle cells of female rabbits were grown, expanded in culture, and characterized immunocytochemically. Vaginal epithelial and smooth muscle cells were seeded on polyglycolic acid (PGA) scaffolds at 10 x 10(6) and 20 x 10(6) cells/cm(3), respectively. The cell-seeded scaffolds were subcutaneously implanted into nude mice. The animals were killed 1, 4, and 6 weeks after implantation. Immunocytochemical and histochemical analyses were performed with pancytokeratins AE1/AE3 and with smooth muscle specific alpha-actin antibodies to confirm the reconstituted tissue phenotype. Western blot analyses and electrical field stimulation studies were also performed to further characterize the tissue-engineered constructs. Vaginal epithelial cells were serially identified with anti-pancytokeratins AE1/AE3 at all culture stages. Smooth muscle cells in culture stained positively with alpha smooth muscle actin antibodies. One week after implantation in vivo, the retrieved polymer scaffolds demonstrated multilayered tissue strips of both cell types, and penetrating native vasculature was also noted. Increased organization of the smooth muscle and epithelial tissue was evident by 4 weeks. There was no evidence of tissue formation in the controls. Immunocytochemical analyses using anti-pancytokeratins confirmed the presence of vaginal epithelial cells in each of the constructs. Anti-alpha-actin smooth muscle antibodies also confirmed the presence of multilayered smooth muscle fibers and tissue at each time point. Western blot analyses of the scaffolds confirmed the expression of cytokeratin and smooth muscle actin proteins when compared with controls. The contractile properties of the tissue-engineered vaginal constructs in response to electrical field stimulation were similar to those of normal vaginal tissue. Vaginal epithelial and smooth muscle cells can be easily cultured and expanded in vitro. Cell-seeded polymer scaffolds are able to form vascularized vaginal tissue in vivo that have phenotypic and functional properties similar to those of normal vaginal tissues. This is the first demonstration in tissue engineering wherein vaginal epithelial and smooth muscle cells are reconstituted in vivo into vaginal tissue. This technology may be pursued further experimentally in order to achieve the engineering of vaginal tissues for clinical applications. PMID- 12740093 TI - Small intestinal submucosa induces loss of mitochondrial integrity and caspase dependent apoptosis in human T cells. AB - Porcine small intestinal submucosa (SIS) is a cell-free biomaterial used in humans for wound healing and as scaffold material for constructive remodeling of damaged or missing tissue. We have previously shown that SIS contains a factor that suppresses human helper T cell subset differentiation and expansion by inducing programmed cell death. Our aims here were to identify in detail the processes involved in SIS-induced T cell apoptosis and to perform the first characterization of the apoptosis-inducing factor present in SIS. In in vitro experiments, we utilized human T cell lines, Jurkat and CEM, to identify the processes involved in SIS-induced T cell apoptosis. Two types of sterile SIS material were used: hydrated sheets and rehydrated clinical-grade sheets. We found that SIS-mediated apoptosis as detected by induction of membrane annexin V staining involved the loss of mitochondrial membrane potential and was dependent on caspase activation. We eliminated transforming growth factor beta (TGF-beta), Fas ligand (FasL), and galectin family members as factors in SIS-mediated T cell apoptosis. We further established that processes required to prepare SIS for clinical use, freeze-drying, and gas sterilization destroyed the apoptosis inducing factor. SIS contains a factor that induces loss of mitochondrial integrity and caspase-dependent apoptosis in human T cells. This factor is destroyed by freeze-drying and gas sterilization and is not TGF-beta, FasL, or a galectin family member. Normal T cell homeostasis in gut-associated tissues may be regulated in part by this unknown factor. PMID- 12740094 TI - Evaluation of various seeding techniques for culturing osteogenic cells on titanium fiber mesh. AB - The objective of the present study was to learn more about the effect of seeding and loading techniques on the osteogenic differentiation in vitro of rat bone marrow cells into titanium fiber mesh. This material was used as received or subjected to glow discharge treatment (RFGD). The seeding methods that were used included a so-called droplet, cell suspension (high and low cell density), and rotating plate method. Osteogenic cells were cultured for 4, 8, and 16 days into titanium fiber mesh. DNA, osteocalcin, scanning electron microscopy (SEM) analysis, and calcium measurements were used to determine cellular proliferation and differentiation. DNA analysis of the differently seeded specimens showed that proliferation proceeded faster in the first versus second run for droplet and cell suspension samples. No clear and distinct additional effect was found when RFGD treatment was used. Statistical analyses revealed that high cell density and low rotational speed resulted always in a significantly higher DNA content. Calcium measurements and osteocalcin analysis showed that using high cell densities during inoculation of the scaffolds prevented the occurrence of differences between experimental runs. SEM examination showed that for droplet and cell suspension samples cells were present at only one side of the mesh. The mesh side where the cell sheet was observed depended on the additional use of glow discharge treatment. On these materials, the cells had penetrated through the meshes and formed a cell sheet at the bottom side. When rotation was used, no cell sheet was formed and cells had invaded the meshes and were growing around the titanium fibers. On the basis of our results, we conclude that (1). titanium fiber mesh is indeed suitable to support the osteogenic expression of bone marrow cells, and (2). changing the initial cell density as well as the use of dynamic seeding methods can influence the osteogenic capacity of the scaffold. PMID- 12740095 TI - Viable osteogenic cells are obligatory for tissue-engineered ectopic bone formation in goats. AB - In this study we investigated the bone-forming capacity of tissue-engineered (TE) constructs implanted ectopically in goats. As cell survival is questionable in large animal models, we investigated the significance of vitality, and thus whether living cells instead of only the potentially osteoinductive extracellular matrix are required to achieve bone formation. Vital TE constructs of porous hydroxyapatite (HA) covered with differentiated bone marrow stromal cells (BMSCs) within an extracellular matrix (ECM) were compared with identical constructs that were devitalized before implantation. The devitalized implants did contain the potentially osteoinductive ECM. Furthermore, we evaluated HA impregnated with fresh bone marrow and HA only. Two different types of HA granules with a volume of approximately 40 microm were investigated: HA70/800, a microporous HA with 70% interconnected macroporosity and an average pore size of 800 microm, and HA60/400, a smooth HA with 60% interconnected macropores and an average size of 400 microm. Two granules of each type were combined and then treated as a single unit for cell seeding, implantation, and histology. The tissue-engineered samples were obtained by seeding culture-expanded goat BMSCs on the HA and subsequently culturing these constructs for 6 days to allow cell differentiation and ECM formation. To devitalize, TE constructs were frozen in liquid nitrogen according to a validated protocol. Fresh bone marrow impregnation was performed perioperatively (4 mL per implant unit). All study groups were implanted in bilateral paraspinal muscles. Fluorochromes were administered at three time points to monitor bone mineralization. After 12 weeks the units were explanted and analyzed by histology of nondecalcified sections. Bone formation was present in all vital tissue-engineered implants. None of the other groups showed any bone formation. Histomorphometry indicated that microporous HA70/800 yielded more bone than did HA60/400. Within the newly formed bone, the fluorescent labels showed that mineralization had occurred before 5 weeks of implantation and was directed from the HA surface toward the center of the pores. In conclusion, tissue engineered bone formation in goats can be achieved only with viable constructs of an appropriate scaffold and sufficient BMSCs. PMID- 12740096 TI - Calcium phosphate-chitosan composite scaffolds for bone tissue engineering. AB - Macroporous calcium phosphate-chitosan composite scaffolds were fabricated and evaluated for use in bone tissue engineering. Human osteoblast-like MG63 cells were cultured on the composite scaffolds, and their response to the materials was studied. Cell morphology, total protein content, and expression of classic markers for osteoblast differentiation were characterized. MG63 cells on the hydroxyapatite scaffolds nesting chitosan sponges (HC1) showed significantly higher alkaline phosphatase (ALP) level and osteocalcin (OC) production during the 11-day culture period, compared with the control culture on tissue culture plates. Cells on the chitosan scaffolds incorporated with hydroxyapatite powders (HC2) exhibited lower ALP activity during the 11-day culture period and OC secretion during the first 7 days, in comparison with that on HC1. The addition of calcium phosphate glass as in HC3 scaffolds increased the ALP and OC levels of MG63 cells. Our study indicated that the hydroxyapatite-matrix composite scaffolds might enhance the phenotype expression of MG63 cells, in comparison with chitosan-matrix scaffolds. Soluble calcium phosphate glasses should be added to the scaffolds to prevent chitosan from fast degradation that may affect the differentiation of osteoblast cells. PMID- 12740097 TI - Human mesenchymal stem cells transduced with recombinant bone morphogenetic protein-9 adenovirus promote osteogenesis in rodents. AB - The present study was undertaken to determine whether ex vivo bone morphogenetic protein-9 (BMP-9) gene therapy using human mesenchymal stem cells (hMSCs) can induce endochondral bone formation in athymic nude rats. An in vitro study was initially performed on hMSCs to evaluate morphological changes and osteoblastic differentiation induced by replication-defective adenovirus type 5 with the cytomegalovirus promoter and either the BMP-9 (Ad-BMP-9) or beta-galactosidase (Ad-beta-gal) gene. In vivo, athymic nude rats received an injection (10(6) hMSCs transduced with recombinant adenovirus at 50 PFU/cell) into the anterior thigh musculature: Ad-BMP-9 on the left and Ad-beta-gal (control) on the right. Computed tomography scans and histological analysis were obtained 7, 14, 28, 42, 56, and 84 days postinjection. In vitro, human mesenchymal stem cells treated with Ad-BMP-9 (50 PFU/cell) showed signs of differentiation, whereas hMSCs treated with 250 and 1250 PFU/cell showed cytotoxicity. In vivo, computed tomography and histological analysis clearly demonstrated ectopic bone at hMSC/Ad BMP-9 treatment sites, whereas the hMSC/Ad-beta-gal treatment sites showed no evidence of osteogenesis. None of the animals showed clinical evidence of toxicity. Ex vivo gene therapy with hMSC/BMP-9 may be efficacious for promoting bone formation for a variety of bone pathologies and certainly warrants further investigations. PMID- 12740098 TI - Formation of hydroxyapatite-mediated three-dimensional structures by mouse fibroblasts in response to physical stimulation. AB - Hydroxyapatite (HAP) ceramics are widely used as implant materials for periodontal bone defects because of their excellent biocompatibility. We demonstrated that physical stimulation, that is, (1). mechanical stimuli or (2). laser irradiation, causes HAP-mediated C3H10T1/2 mouse fibroblasts to form three dimensional tissue-like structures. Trypsinized 10T1/2 cells were cultured simultaneously with 200 HAP granules on a rotator for 7 days in mechanical stimulation experiments. The cells were later transferred to a regular incubator. Cell reactions were observed by phase-contrast microscopy. The formation of three dimensional structures around the HAP granules was observed in the third week of cultivation after stimulation. In laser irradiation experiments, trypsinized cells were irradiated with 1, 5, and 16 J/cm(2) at a wavelength of 1000 nm and cultured with 200 HAP granules for 10 weeks. The formation of three-dimensional structures, like those observed in the mechanical stimulation experiments, was observed in the third week after irradiation. The formation of these structures was most frequent at 1 J/cm(2), and the frequency of formation of these structures gradually decreased as the irradiation dose was increased. These results indicate that physical stimuli may stimulate cell proliferation, leading to the repair of damaged tissue. These results also indicate that mouse fibroblasts do not form these three-dimensional structures without HAP and that HAP alone is not sufficient to stimulate the formation of three-dimensional structures. PMID- 12740099 TI - Tissue engineering by cocultivating human elastic chondrocytes and keratinocytes. AB - To date, there is no optimal way to reconstruct an external ear in cases of microtia or after trauma or burns damaging the external ear. However, success in the area of tissue engineering has indicated that autologous elastic cartilage produced in vitro might be of great importance in the future treatment of these patients. In the present study we have engineered human, elastic cartilage in vitro by culturing chondrocytes in fibrin glue. Furthermore, the engineered elastic cartilage was seeded with human keratinocytes to investigate the possibility of combining these two tissues into one integrated structure. Histological analysis and immunohistochemistry were done every second week for 10 weeks. The elastic chondrocytes were shown to grow well in the matrix and proliferated in a dense pattern. After 10 weeks a matrix containing elastin was shown by staining with orcein, indicating that an elastic cartilage had been formed. The seeded keratinocytes adhered to the cartilage, proliferated, and formed a stratified epidermal layer, which was shown by routine histological staining and immunohistochemistry. This study shows that human elastic chondrocytes can be cultured in fibrin glue and that human keratinocytes can be cocultured with this engineered cartilage, which might be of great importance in future reconstruction of ears. PMID- 12740100 TI - Tissue-engineered cartilage using an injectable and in situ gelable thermoresponsive gelatin: fabrication and in vitro performance. AB - An injectable and in situ gelable scaffold can fully fill the space of cartilaginous defects of complex shapes. The authors attempted to develop a novel injection-driven technique for cartilage repair using a thermoresponsive gelatin, poly(N-isopropylacrylamide)-grafted gelatin (PNIPAAm-gelatin). A mixed solution of chondrocytes was isolated from a Japanese white rabbit and PNIPAAm-gelatin was spontaneously solidified at 37 degrees C and cultured. The number of cells in the gel with a poly(N-isopropylacrylamide) (PNIPAAm) chain of high molecular weight (1.3 x 10(5) g/mol) and at low concentration (5 w/v%) remained unchanged irrespective of culture time, and minimal cell death and little cell proliferation were observed. A round-shaped morphology was dominantly restored even at 1 week of incubation. The cell population in the G(0)/G(1) phase was high (more than 90%), and this gradually increased with culture time. Type II collagen and sulfated glycosaminoglycan (s-GAG) were detected in the tissue-engineered cartilage, but a small amount of type I collagen was also detected. Total collagen and s-GAG increased in level close to those of native hyaline cartilage over 12 weeks of culture. Mechanical properties of the tissue-engineered cartilage responding to loading and unloading of compression force tend to approach those of native hyaline cartilage with culture time. These results suggest that PNIPAAm-gelatin may be a suitable in situ formable scaffold for cartilage repair. PMID- 12740101 TI - Federal regulation in the absence of prescription drug legislation: the Bush administration's proposal to speed generics to market. PMID- 12740102 TI - Introduction. Long-term potentiation and structure of the issue. PMID- 12740103 TI - A prelude to long-term potentiation. AB - Searching for premonitory studies of hippocampal long-term potentiation (LTP), there is a paucity of data. While synaptic enhancement during repetitive activation was studied in several reports from many groups between 1955 and 1967, the reported after-effects were short, at the most lasting a few minutes. Responses lasting for more than 1 hour were not reported until 1973. PMID- 12740104 TI - The discovery of long-term potentiation. AB - This paper describes circumstances around the discovery of long-term potentiation (LTP). In 1966, I had just begun independent work for the degree of Dr medicinae (PhD) in Per Andersen's laboratory in Oslo after an eighteen-month apprenticeship with him. Studying the effects of activating the perforant path to dentate granule cells in the hippocampus of anaesthetized rabbits, I observed that brief trains of stimuli resulted in increased efficiency of transmission at the perforant path-granule cell synapses that could last for hours. In 1968, Tim Bliss came to Per Andersen's laboratory to learn about the hippocampus and field potential recording for studies of possible memory mechanisms. The two of us then followed up my preliminary results from 1966 and did the experiments that resulted in a paper that is now properly considered to be the basic reference for the discovery of LTP. PMID- 12740105 TI - A journey from neocortex to hippocampus. AB - In the mid-1960s, it was generally agreed that the engram, the neural trace of previously experienced events, must be encoded by Hebb-like neurons in which synaptic efficacy could be modified by activity. Here, I describe my attempts as a PhD student at McGill University, Montreal, to find rules governing cortical plasticity in the neocortex, and having failed, why the hippocampus seemed to offer a far better prospect. PMID- 12740106 TI - Long-term potentiation in the Eocene. AB - The first ten years of long-term potentiation (LTP) research are reviewed. Surprisingly, given the intensity of current interest, the discovery paper did not trigger a wave of follow-on experiments. Despite this, the initial work laid out what ultimately became standard questions and paradigms. The application of the then still novel hippocampal slice technique oriented LTP towards basic neuroscience, perhaps somewhat at the cost of lesser attention to its functional significance. The use of slices led to the discovery of the events that trigger the formation of LTP and provided some first clues about its extraordinary persistence. Signs of the intense controversy over the nature of LTP expression (release vs receptors) emerged towards the end of the first decade of work. What appears to be lacking in the literature of that time is a widespread concern about LTP and memory. This may reflect a somewhat different attitude that neurobiologists then had towards memory research and a perceived need to integrate the new potentiation phenomenon into the web of established science before advancing extended arguments about its contributions to behaviour. PMID- 12740107 TI - Long-term potentiation, cooperativity and Hebb's cell assemblies: a personal history. AB - The early history of the experimental work leading to the discovery that long term potentiation (LTP) embodies Hebb's principle of association is described. In addition, the fallacy underlying the sometimes presumed distinction between 'cooperativity' and 'associativity' in the induction of LTP is pointed out. PMID- 12740108 TI - The induction of N-methyl-D-aspartate receptor-dependent long-term potentiation. AB - The role of N-methyl-D-aspartate (NMDA) receptors in the induction of long-term potentiation (LTP) was established during the 1980s. In this article I present a personal reflection upon the role that my colleagues and I played in the discovery of the mechanism of induction of NMDA receptor-dependent LTP. PMID- 12740109 TI - Long-term potentiation and memory. AB - The discovery of long-term potentiation (LTP) transformed research on the neurobiology of learning and memory. This did not happen overnight, but the discovery of an experimentally demonstrable phenomenon reflecting activity-driven neuronal and synaptic plasticity changed discussions about what might underlie learning from speculation into something much more concrete. Equally, however, the relationship between the discovery of LTP and research on the neurobiology of learning and memory has been reciprocal; for it is also true that studies of the psychological, anatomical and neurochemical basis of memory provided a developing and critical intellectual context for the physiological discovery. The emerging concept of multiple memory systems, from 1970 onwards, paved the way for the development of new behavioural and cognitive tasks, including the watermaze described in this paper. The use of this task in turn provided key evidence that pharmacological interference with an LTP induction mechanism would also interfere with learning, a finding that was by no means a foregone conclusion. This reciprocal relationship between studies of LTP and the neurobiology of memory helped the physiological phenomenon to be recognized as a major discovery. PMID- 12740110 TI - Bidirectional synaptic plasticity: from theory to reality. AB - Theories of receptive field plasticity and information storage make specific assumptions for how synapses are modified. I give a personal account of how testing the validity of these assumptions eventually led to a detailed understanding of long-term depression and metaplasticity in hippocampal area CA1 and the visual cortex. The knowledge of these molecular mechanisms now promises to reveal when and how sensory experience modifies synapses in the cerebral cortex. PMID- 12740111 TI - Kainate receptors and the induction of mossy fibre long-term potentiation. AB - There is intense interest in understanding the molecular mechanisms involved in long-term potentiation (LTP) in the hippocampus. Significant progress in our understanding of LTP has followed from studies of glutamate receptors, of which there are four main subtypes (alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA), N-methyl-D-aspartate (NMDA), mGlu and kainate). This article summarizes the evidence that the kainate subtype of glutamate receptor is an important trigger for the induction of LTP at mossy fibre synapses in the CA3 region of the hippocampus. The pharmacology of the first selective kainate receptor antagonists, in particular the GLU(K5) subunit selective antagonist LY382884, is described. LY382884 selectively blocks the induction of mossy fibre LTP, in response to a variety of different high-frequency stimulation protocols. This antagonist also inhibits the pronounced synaptic facilitation of mossy fibre transmission that occurs during high-frequency stimulation. These effects are attributed to the presence of presynaptic GLU(K5)-subunit-containing kainate receptors at mossy fibre synapses. Differences in kainate receptor-dependent synaptic facilitation of AMPA and NMDA receptor-mediated synaptic transmission are described. These data are discussed in the context of earlier reports that glutamate receptors are not involved in mossy fibre LTP and more recent experiments using kainate receptor knockout mice, that argue for the involvement of GLU(K6) but not GLU(K5) kainate receptor subunits. We conclude that activation of presynaptic GLU(K5)-containing kainate receptors is an important trigger for the induction of mossy fibre LTP in the hippocampus. PMID- 12740113 TI - Long-term potentiation in the dentate gyrus of the anaesthetized rat is accompanied by an increase in extracellular glutamate: real-time measurements using a novel dialysis electrode. AB - We have used a glutamate-specific dialysis electrode to obtain real-time measurements of changes in the concentration of glutamate in the extracellular space of the hippocampus during low-frequency stimulation and following the induction of long-term potentiation (LTP). In the dentate gyrus, stimulation of the perforant path at 2 Hz for 2 min produced a transient increase in glutamate current relative to the basal value at control rates of stimulation (0.033 Hz). This activity-dependent glutamate current was significantly enhanced 35 and 90 min after the induction of LTP. The maximal 2 Hz signal was obtained during post tetanic potentiation (PTP). There was also a more gradual increase in the basal level of extracellular glutamate following the induction of LTP. Both the basal and activity-dependent increases in glutamate current induced by tetanic stimulation were blocked by local infusion of the N-methyl-D-aspartate receptor antagonist D-APV. In areas CA1 and CA3 we were unable to detect a 2 Hz glutamate signal either before or after the induction of LTP, possibly owing to a more avid uptake of glutamate in the pyramidal cell fields. These results demonstrate that LTP in the dentate gyrus is associated with a greater concentration of extracellular glutamate following activation of potentiated synapses, either because potentiated synapses release more transmitter per impulse, or because of reduced uptake by glutamate transporters. We present arguments favouring increased release rather than decreased uptake. PMID- 12740112 TI - Active dendrites, potassium channels and synaptic plasticity. AB - The dendrites of CA1 pyramidal neurons in the hippocampus express numerous types of voltage-gated ion channel, but the distributions or densities of many of these channels are very non-uniform. Sodium channels in the dendrites are responsible for action potential (AP) propagation from the axon into the dendrites (back propagation); calcium channels are responsible for local changes in dendritic calcium concentrations following back-propagating APs and synaptic potentials; and potassium channels help regulate overall dendritic excitability. Several lines of evidence are presented here to suggest that back-propagating APs, when coincident with excitatory synaptic input, can lead to the induction of either long-term depression (LTD) or long-term potentiation (LTP). The induction of LTD or LTP is correlated with the magnitude of the rise in intracellular calcium. When brief bursts of synaptic potentials are paired with postsynaptic APs in a theta-burst pairing paradigm, the induction of LTP is dependent on the invasion of the AP into the dendritic tree. The amplitude of the AP in the dendrites is dependent, in part, on the activity of a transient, A-type potassium channel that is expressed at high density in the dendrites and correlates with the induction of the LTP. Furthermore, during the expression phase of the LTP, there are local changes in dendritic excitability that may result from modulation of the functioning of this transient potassium channel. The results support the view that the active properties of dendrites play important roles in synaptic integration and synaptic plasticity of these neurons. PMID- 12740114 TI - Imaging spatio-temporal patterns of long-term potentiation in mouse hippocampus. AB - Spatio-temporal patterns of neuronal activity before and after the induction of long-term potentiation in mouse hippocampal slices were studied using a real-time high-resolution optical recording system. After staining the slices with voltage sensitive dye, transmitted light images and extracellular field potentials were recorded in response to stimuli applied to CA1 stratum radiatum. Optical and electrical signals in response to single test pulses were enhanced for at least 30 minutes after brief high-frequency stimulation at the same site. In two pathway experiments, potentiation was restricted to the tetanized pathway. The optical signals demonstrated that both the amplitude and area of the synaptic response were increased, in patterns not predictable from the initial, pretetanus, pattern of activation. Optical signals will be useful for investigating spatio-temporal patterns of synaptic enhancement underlying information storage in the brain. PMID- 12740115 TI - Fusion pore modulation as a presynaptic mechanism contributing to expression of long-term potentiation. AB - Working on the idea that postsynaptic and presynaptic mechanisms of long-term potentiation (LTP) expression are not inherently mutually exclusive, we have looked for the existence and functionality of presynaptic mechanisms for augmenting transmitter release in hippocampal slices. Specifically, we asked if changes in glutamate release might contribute to the conversion of 'silent synapses' that show N-methyl-D-aspartate (NMDA) responses but no detectable alpha amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) responses, to ones that exhibit both. Here, we review experiments where NMDA receptor responses provided a bioassay of cleft glutamate concentration, using opposition between peak [glu](cleft )and a rapidly reversible antagonist, L-AP5. We discuss findings of a dramatic increase in peak [glu](cleft) upon expression of pairing-induced LTP (Choi). We present simulations with a quantitative model of glutamatergic synaptic transmission that includes modulation of the presynaptic fusion pore, realistic cleft geometry and a distributed array of postsynaptic receptors and glutamate transporters. The modelling supports the idea that changes in the dynamics of glutamate release can contribute to synaptic unsilencing. We review direct evidence from Renger et al., in accord with the modelling, that trading off the strength and duration of the glutamate transient can markedly alter AMPA receptor responses with little effect on NMDA receptor responses. An array of additional findings relevant to fusion pore modulation and its proposed contribution to LTP expression are considered. PMID- 12740117 TI - GluR2 protein-protein interactions and the regulation of AMPA receptors during synaptic plasticity. AB - AMPA-type glutamate receptors mediate most fast excitatory synaptic transmissions in the mammalian brain. They are critically involved in the expression of long term potentiation and long-term depression, forms of synaptic plasticity that are thought to underlie learning and memory. A number of synaptic proteins have been identified that interact with the intracellular C-termini of AMPA receptor subunits. Here, we review recent studies and present new experimental data on the roles of these interacting proteins in regulating the AMPA receptor function during basal synaptic transmission and plasticity. PMID- 12740116 TI - AMPA receptor trafficking and long-term potentiation. AB - Activity-dependent changes in synaptic function are believed to underlie the formation of memories. A prominent example is long-term potentiation (LTP), whose mechanisms have been the subject of considerable scrutiny over the past few decades. I review studies from our laboratory that support a critical role for AMPA receptor trafficking in LTP and experience-dependent plasticity. PMID- 12740118 TI - Expression mechanisms underlying long-term potentiation: a postsynaptic view. AB - This review summarizes the various experiments that have been carried out to determine if the expression of long-term potentiation (LTP), in particular N methyl-D-aspartate (NMDA) receptor-dependent LTP, is presynaptic or postsynaptic. Evidence for a presynaptic expression mechanism comes primarily from experiments reporting that glutamate overflow is increased during LTP and from experiments showing that the failure rate decreases during LTP. However, other experimental approaches, such as monitoring synaptic glutamate release by recording astrocytic glutamate transporter currents, have failed to detect any change in glutamate release during LTP. In addition, the discovery of silent synapses, in which LTP rapidly switches on alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor function at NMDA-receptor-only synapses, provides a postsynaptic mechanism for the decrease in failures during LTP. It is argued that the preponderance of evidence favours a postsynaptic expression mechanism, whereby NMDA receptor activation results in the rapid recruitment of AMPA receptors as well as a covalent modification of synaptic AMPA receptors. PMID- 12740119 TI - Silent synapses: what are they telling us about long-term potentiation? AB - At several cortical synapses glutamate release events can be mediated exclusively by NMDA receptors, with no detectable contribution from AMPA receptors. This observation was originally made by comparing the trial-to-trial variability of the two components of synaptic signals evoked in hippocampal neurons, and was subsequently confirmed by recording apparently pure NMDA receptor-mediated EPSCs with stimulation of small numbers of axons. It has come to be known as the 'silent synapse' phenomenon, and is widely assumed to be caused by the absence of functional AMPA receptors, which can, however, be recruited into the postsynaptic density by long-term potentiation (LTP) induction. Thus, it provides an important impetus for relating AMPA receptor trafficking mechanisms to the expression of LTP, a theme that is taken up elsewhere in this issue. This article draws attention to several findings that call for caution in identifying silent synapses exclusively with synapses without AMPA receptors. In addition, it attempts to identify several missing pieces of evidence that are required to show that unsilencing of such synapses is entirely accounted for by insertion of AMPA receptors into the postsynaptic density. Some aspects of the early stages of LTP expression remain open to alternative explanations. PMID- 12740120 TI - How long will long-term potentiation last? AB - The paramount feature of long-term potentiation (LTP) as a memory mechanism is its characteristic persistence over time. Although the basic phenomenology of LTP persistence was established 30 years ago, new insights have emerged recently about the extent of LTP persistence and its regulation by activity and experience. Thus, it is now evident that LTP, at least in the dentate gyrus, can either be decremental, lasting from hours to weeks, or stable, lasting months or longer. Although mechanisms engaged during the induction of LTP regulate its subsequent persistence, the maintenance of LTP is also governed by activity patterns post-induction, whether induced experimentally or generated by experience. These new findings establish dentate gyrus LTP as a useful model system for studying the mechanisms governing the induction, maintenance and interference with long-term memory, including very long-term memory lasting months or longer. The challenge is to study LTP persistence in other brain areas, and to relate, if possible, the properties and regulation of LTP maintenance to these same properties of the information that is actually stored in those regions. PMID- 12740121 TI - Structural changes at dendritic spine synapses during long-term potentiation. AB - Two key hypotheses about the structural basis of long-term potentiation (LTP) are evaluated in light of new findings from immature rat hippocampal slices. First, it is shown why dendritic spines do not split during LTP. Instead a small number of spine-like dendritic protrusions may emerge to enhance connectivity with potentiated axons. These 'same dendrite multiple synapse boutons' provide less than a 3% increase in connectivity and do not account for all of LTP or memory, as they do not accumulate during maturation. Second, polyribosomes in dendritic spines served to identify which of the existing synapses enlarged to sustain more than a 30% increase in synaptic strength. Thus, both enhanced connectivity and enlarged synapses result during LTP, with synapse enlargement being the greater effect. PMID- 12740122 TI - Cadherins and synaptic plasticity: activity-dependent cyclin-dependent kinase 5 regulation of synaptic beta-catenin-cadherin interactions. AB - Cyclin-dependent kinase 5 (Cdk5)/p35 kinase activity is known to decrease the affinity of beta-catenin for cadherin in developing cortical neurons. Our recent work demonstrated that depolarization causes an increased affinity between beta catenin and cadherin. Here, we examine whether Cdk5/p35 regulates beta-catenin cadherin affinity in response to neural activity. In hippocampal neurons depolarization caused a significant decrease in Cdk5 kinase activity, without changing the protein levels of either Cdk5 or p35, suggesting that the proteasome pathway is not involved. Decreasing Cdk5 kinase activity with the inhibitor roscovitine increased the amount of beta-catenin that was co-immunoprecipitated with cadherin. Inhibiting Cdk5 activity also resulted in a redistribution of EGFP beta-catenin from the dendritic shaft to the spines, where cadherins are highly concentrated. The redistribution of beta-catenin induced by roscovitine is similar to that induced by depolarization. Interestingly, the redistribution induced by the Cdk5 inhibitor was completely blocked by either a tyrosine phosphatase inhibitor, orthovanadate or by point mutations of beta-catenin Tyr 654 to Glu or Phe. Immunoprecipitation studies further revealed that roscovitine increases the affinity of the wild-type, but not mutated, EGFP-beta-catenin for cadherin. These results suggest that Cdk5 activity regulates the affinity of beta catenin for cadherin by changing the phosphorylation level of beta-catenin Tyr 654. PMID- 12740123 TI - In search of general mechanisms for long-lasting plasticity: Aplysia and the hippocampus. AB - Long-term synaptic plasticity is thought to underlie many forms of long-lasting memory. Long-lasting plasticity has been most extensively studied in the marine snail Aplysia and in the mammalian hippocampus, where Bliss and Lomo first described long-term potentiation 30 years ago. The molecular mechanisms of plasticity in these two systems have proven to have many similarities. Here, we briefly describe some of these areas of overlap. We then summarize recent advances in our understanding of the mechanisms of long-lasting synaptic facilitation in Aplysia and suggest that these may prove fruitful areas for future investigation in the mammalian hippocampus and at other synapses in the mammalian brain. PMID- 12740124 TI - Long-term potentiation and the ageing brain. AB - Ageing is associated with learning and memory impairments. Data are reviewed that suggest that age-related impairments of hippocampal-dependent forms of memory, may be caused, in part, by altered synaptic plasticity mechanisms in the hippocampus, including long-term potentiation (LTP). To the extent that the mechanisms responsible for LTP can be understood, it may be possible to develop therapeutic approaches to alleviate memory decline in normal ageing. PMID- 12740125 TI - Genetic neuroscience of mammalian learning and memory. AB - Our primary research interest is to understand the molecular and cellular mechanisms on neuronal circuitry underlying the acquisition, consolidation and retrieval of hippocampus-dependent memory in rodents. We study these problems by producing genetically engineered (i.e. spatially targeted and/or temporally restricted) mice and analysing these mice by multifaceted methods including molecular and cellular biology, in vitro and in vivo physiology and behavioural studies. We attempt to identify deficits at each of the multiple levels of complexity in specific brain areas or cell types and deduce those deficits that underlie specific learning or memory. We will review our recent studies on the acquisition, consolidation and recall of memories that have been conducted with mouse strains in which genetic manipulations were targeted to specific types of cells in the hippocampus or forebrain of young adult mice. PMID- 12740126 TI - Inducible molecular switches for the study of long-term potentiation. AB - This article reviews technical and conceptual advances in unravelling the molecular bases of long-term potentiation (LTP), learning and memory using genetic approaches. We focus on studies aimed at testing a model suggesting that protein kinases and protein phosphatases balance each other to control synaptic strength and plasticity. We describe how gene 'knock-out' technology was initially exploited to disrupt the Ca(2+)/calmodulin-dependent protein kinase IIalpha (CaMKIIalpha) gene and how refined knock-in techniques later allowed an analysis of the role of distinct phosphorylation sites in CaMKII. Further to gene recombination, regulated gene expression using the tetracycline-controlled transactivator and reverse tetracycline-controlled transactivator systems, a powerful new means for modulating the activity of specific molecules, has been applied to CaMKIIalpha and the opposing protein phosphatase calcineurin. Together with electro-physiological and behavioural evaluation of the engineered mutant animals, these genetic methodologies have helped gain insight into the molecular mechanisms of plasticity and memory. Further technical developments are, however, awaited for an even higher level of finesse. PMID- 12740128 TI - Synaptic plasticity in the mesolimbic dopamine system. AB - Long-term potentiation (LTP) and long-term depression (LTD) are thought to be critical mechanisms that contribute to the neural circuit modifications that mediate all forms of experience-dependent plasticity. It has, however, been difficult to demonstrate directly that experience causes long-lasting changes in synaptic strength and that these mediate changes in behaviour. To address these potential functional roles of LTP and LTD, we have taken advantage of the powerful in vivo effects of drugs of abuse that exert their behavioural effects in large part by acting in the nucleus accumbens (NAc) and ventral tegmental area (VTA); the two major components of the mesolimbic dopamine system. Our studies suggest that in vivo drugs of abuse such as cocaine cause long-lasting changes at excitatory synapses in the NAc and VTA owing to activation of the mechanisms that underlie LTP and LTD in these structures. Thus, administration of drugs of abuse provides a distinctive model for further investigating the mechanisms and functions of synaptic plasticity in brain regions that play important roles in the control of motivated behaviour, and one with considerable practical implications. PMID- 12740127 TI - MAPK, CREB and zif268 are all required for the consolidation of recognition memory. AB - There has been nearly a century of interest in the idea that encoding and storage of information in the brain requires changes in the efficacy of synaptic connections between neurons that are activated during learning. Recent research into the molecular mechanisms of long-term potentiation (LTP) has brought about new knowledge that has provided valuable insights into the neural mechanisms of memory storage. The evidence indicates that rapid activation of the genetic machinery can be a key mechanism underlying the enduring modification of neural networks required for the stability of memories. In recent years, a wealth of experimental data has highlighted the importance of mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) signalling in the regulation of gene transcription in neurons. Here, we briefly review experiments that have shown MAPK/ERK, cAMP response element-binding protein (CREB) and the immediate early gene (IEG) zif268 are essential components of a signalling cascade required for the expression of late phase LTP and of certain forms of long-term memory. We also present experiments in which we have assessed the role of these three molecules in recognition memory. We show that pharmacological blockade of MAPK/ERK phosphorylation, functional inactivation of CREB in an inducible transgenic mouse and inactivation of zif268 in a mutant mouse result in a similar deficit in long-term recognition memory. In the continuing debate about the role of LTP mechanisms in memory, these findings provide an important complement to the suggestion that synaptic changes brought about by LTP and memory consolidation and storage share, at least in part, common underlying molecular mechanisms. PMID- 12740129 TI - Synaptic plasticity in animal models of early Alzheimer's disease. AB - Amyloid beta-protein (Abeta) is believed to be a primary cause of Alzheimer's disease (AD). Recent research has examined the potential importance of soluble species of Abeta in synaptic dysfunction, long before fibrillary Abeta is deposited and neurodegenerative changes occur. Hippocampal excitatory synaptic transmission and plasticity are disrupted in transgenic mice overexpressing human amyloid precursor protein with early onset familial AD mutations, and in rats after exogenous application of synthetic Abeta both in vitro and in vivo. Recently, naturally produced soluble Abeta was shown to block the persistence of long-term potentiation (LTP) in the intact hippocampus. Sub-nanomolar concentrations of oligomeric Abeta were sufficient to inhibit late LTP, pointing to a possible reason for the sensitivity of hippocampus-dependent memory to impairment in the early preclinical stages of AD. Having identified the active species of Abeta that can play havoc with synaptic plasticity, it is hoped that new ways of targeting early AD can be developed. PMID- 12740131 TI - Leptin and insulin action in severely obese women. AB - BACKGROUND: The authors investigated the interrelationships between the components of the metabolic syndrome in severe obesity. METHODS: In non-diabetic, severely obese women, the degree of obesity (BMI), the insulin sensitivity (from the Homeostatic Model of Assessment, HOMA), the serum leptin concentration and the presence of dyslipidemia and arterial hypertension were evaluated. RESULTS: In insulin-resistant patients, an overall impaired metabolic status and a greater cardiovascular risk were observed, while serum leptin concentration was higher than in the insulin-sensitive ones. Leptin levels and HOMA data correlated independent of BMI findings, while the presence of dyslipidemia and hypertension was unrelated to the other metabolic syndrome factors. CONCLUSION: In severely obese women, although other factors independently intervene, serum leptin has a role in developing the metabolic syndrome. PMID- 12740132 TI - The impact of bariatric surgery on the Veterans Administration healthcare system: a cost analysis. AB - BACKGROUND: The economic burden of caring for veterans with clinically severe obesity and its comorbidities is straining the Veterans Administration (VA) healthcare system. The authors determined the cost of Roux-en-Y Gastric Bypass (RYGBP) in the VA's single-payor healthcare system. METHODS: The records of all 25 patients who underwent RYGBP from May 1999 to October 2001 were reviewed. All obesity-related health-care costs including hospitalizations as well as outpatient visits, medications and home health devices were calculated for 12 months before and after the RYGBP. RESULTS: Age was 52+/-2 yr and preoperative BMI was 52+/-2 kg/m(2); ASA score was III (21 patients) and II (4 patients). Mean follow-up was 18 months. Total cost of care for these patients preoperatively was $10,778+/-2,460/patient (outpatient visits=$5,476+/-682, hospital admissions=$12,221+/-6,062, and home health devices=$1,383+/-349). Postoperative length of stay was 8+/-0.5 days. Cost of the gastric bypass was $8,976+/-497/pt (OR fixed cost=$1,900/patient + ICU and ward=$7,076+/-497/patient). For the first postoperative year, 6 patients had 12 admissions, but routine outpatient visits were significantly reduced from 55+/-6 to 18+/-2 postoperatively (P<0.001). The cost of all care excluding peri-operative charges for 1 year after gastric bypass was $2,840+/-622/patient (P=0.005 vs preop). CONCLUSIONS: Operative treatment of clinically severe obesity reduces obesity-related expenditures and utilization of healthcare resources. The cost of undertaking RYGBP at the VA is offset by reduction of health-care costs within the first year after surgery. These data support allocation of resources to support existing bariatric surgery programs throughout the VA system. PMID- 12740133 TI - Heparin thromboprophylaxis in gastric bypass surgery. AB - BACKGROUND: Patients undergoing gastric bypass surgery are at risk for postoperative venous thromboembolism. Thromboprophylaxis often includes fixed doses of some type of heparin. However, it is unlikely that the same dose of subcutaneous heparin will be optimal for all patients, because heparin pharmacokinetics depend on a number of patient variables, including thickness of the adipose layer. METHODS: An adjusted-dose, unfractionated heparin protocol was developed using pharmacokinetic data from 245 medical and surgical patients. Heparin doses were adjusted to achieve subtherapeutic peak anti-factor Xa heparin activity levels of 0.11-0.25 units/mL. This protocol was then applied to a prospective series of 700 patients undergoing laparoscopic Roux-en-Y gastric bypass who had no history of thromboembolism. Heparin prophylaxis was begun the evening of the day of surgery. RESULTS: No patients were diagnosed with a deep venous thrombosis, but 3 (0.4%) were diagnosed with a non-fatal pulmonary embolism. Heparin therapy was halted because of bleeding in 2.3% of patients but only half of these required blood transfusions (1% of total). No patient required reoperation. Minor wound hematomas occurred in 0.6%. There were no deaths from any cause in this series. CONCLUSION: Use of a monitored, adjusted-dose unfractionated heparin prophylactic protocol in a laparoscopic gastric bypass patient population resulted in doses greater than those used in traditional fixed dose protocols. However, bleeding and thromboembolism rates were very low and no patients died. PMID- 12740134 TI - Treatment of dilated gastrojejunostomy with sclerotherapy. AB - BACKGROUND: Dilation of the gastrojejunostomy after gastric bypass may result in weight gain. Many surgical and medical treatments have met with poor results. A feasibility study of endoscopic sclerotherapy (ST) of the gastrojejunostomy was performed, based on the known risk of esophageal stricture in the treatment of esophageal varices. METHODS: From 1991 to 2001, proximal Roux-en-Y gastric bypass (RYGBP) was performed on 685 patients, with a follow-up rate of 60% at 5 years. 20 patients were identified with dilated gastrojejunostomy (DGJ) by gastroscopy (EGD) performed for complaints of weight gain and marked increase in volume tolerance. Sclerotherapy of the gastrojejunostomy was performed during EGD. EGD was repeated 2 months after the sclerotherapy to measure the diameter of the anastomosis and observe any complications. Weight and sense of satiety were also measured, 2 and 6 months after ST. If necessary, the procedure was repeated to achieve a diameter of 10 mm. RESULTS: Reducing the diameter of the gastrojejunostomy to 9-10 mm was achieved in all patients, with an average of 1.3 treatments per patient. 15 patients (75%) lost weight. The average weight loss was 5.8 kg (ranging from 0.5 to 17.3 kg) in 2 months. CONCLUSION: Sclerotherapy successfully restores the desired anatomy of gastric bypass, but exercise and dietary discretion remain critical elements of sustained weight loss. PMID- 12740130 TI - Long-term potentiation: outstanding questions and attempted synthesis. AB - This article attempts an overview of the mechanism of NMDAR-dependent long-term potentiation (LTP) and its role in hippocampal networks. Efforts are made to integrate information, often in speculative ways, and to identify unresolved issues about the induction, expression and molecular storage processes. The pre/post debate about LTP expression has been particularly difficult to resolve. The following hypothesis attempts to reconcile the available physiological evidence as well as anatomical evidence that LTP increases synapse size. It is proposed that synapses are composed of a variable number of trans-synaptic modules, each having presynaptic release sites and a postsynaptic structure that can be AMPAfied by the addition of a hyperslot assembly that anchors 10-20 AMPA channels. According to a newly developed view of transmission, the quantal response is generated by AMPA channels near the site of vesicle release and so will depend on whether the module where release occurs has been AMPAfied. LTP expression may involve two structurally mediated processes: (i) the AMPAfication of existing modules by addition of hyperslot assemblies: this is a purely postsynaptic process and produces an increase in the probability of an AMPA response, with no change in the NMDA component; and (ii) the addition of new modules: this is a structurally coordinated pre/post process that leads to LTP induced synapse enlargement and potentiation of the NMDA component owing to an increase in the number of release sites (the number of NMDA channels is assumed to be fixed). The protocol used for LTP induction appears to affect the proportion of these two processes; pairing protocols that involve low-frequency presynaptic stimulation induce only AMPAfication, making LTP purely postsynaptic, whereas high-frequency stimulation evokes both processes, giving rise to a presynaptic component. This model is capable of reconciling much of the seemingly contradictory evidence in the pre/post debate. The structural nature of the postulated changes is relevant to a second debate: whether a CaMKII switch or protein-dependent structural change is the molecular memory mechanism. A possible reconciliation is that a reversible CaMKII switch controls the construction of modules and hyperslot assemblies from newly synthesized proteins. PMID- 12740135 TI - Taking posterior rectus sheath laparoscopically to reinforce the gastrojejunostomy in laparoscopic Roux-en-Y gastric bypass. AB - BACKGROUND: The Roux-en-Y gastric bypass (RYGBP) is now performed laparoscopically widely with low morbidity and mortality. However, in some cases long-term adequate weight loss is not satisfied because of dilatation of the gastrojejunostomy. Therefore, a prosthetic material and bio-membranes have been used to prevent dilatation. In this study, we used posterior rectus sheath by laparoscopy, to evaluate feasibility and safety of the procedure. METHODS: 20 Yorkshire pigs, under general anesthesia, had a standard laparoscopic RYGBP. In addition, 10 had their gastrojejunostomy anastomosis wrapped with 2x10 cm posterior rectus sheath. Clinical and operative outcome after operation were compared with the control group of laparoscopic RYGBP cases. RESULTS: The median weight of the pigs was 46.1 kg (range 42-51) in the posterior rectus sheath applied group and 45.2 kg (range 42-49) in the control group. All gastrojejunostomies in the posterior rectus sheath-applied group were successfully reinforced laparoscopically. Both groups loss weight compared with their normal growth weight, but there was no significant difference in the median weight loss between the two groups. Two pigs in the posterior rectus sheath applied group developed a stenosis at the gastrojejunostomy anastomosis following RYGBP. All pigs in the posterior rectus sheath-applied group were found to develop hypertrophic smooth muscle and connective tissue scarring at the gastrojejunostomy on histologic examination. CONCLUSION: Laparoscopic application of posterior rectus sheath around the gastrojejunostomy in laparoscopic RYGBP is feasible and safe. The sheath-applied group developed stenosis and connective tissue scarring. Additional research is needed to evaluate effectiveness in preventing dilation of the anastomosis. PMID- 12740136 TI - Laparoscopic technique for performing duodenal switch with gastric reduction. AB - BACKGROUND: The duodenal switch procedure with gastric reduction (DS) is a hybrid procedure for morbid obesity that combines moderate intake restriction with moderate malabsorption. This report describes the laparoscopic hand-assisted technique for the duodenal switch procedure (LapDS). METHODS: Restriction is achieved via a greater curvature gastrectomy, reducing gastric capacity to 120 ml. The malabsorptive component is constructed by dividing the duodenum 4 cm distal to the pylorus and anastomosing the proximal duodenum to the distal 250 cm of ileum. The biliopancreatic limb is anastomosed to create a 100 cm common channel. Laparoscopic cholecystectomy, cholangiogram, liver biopsy and appendectomy are performed in conjunction with DS. RESULTS: 345 LapDS procedures (27 lap-assisted; 318 hand-assisted) were performed between September 1999 and February 2002. There were 299 women and 46 men with a mean age of 43 years (range 19-67 years). Mean BMI was 50 (range 36-118 kg/m(2)). Mean operating time was 201 minutes (range 105-480). The median length of hospital stay was 3.0 days (range 2 22 days, excluding one outlier). There were 7 conversions to open laparotomy, 14 reoperations, and 21 readmissions. There were 3 pulmonary emboli, 2 deep venous thromboses, and 4 perioperative proximal anastomotic strictures. There were no deaths. Mean percent excess weight loss at 6, 18, and 24 months was 51%, 89%, and 91%, respectively. CONCLUSION: Laparoscopic assisted duodenal switch procedure can be performed safely with acceptable operative times and without excess morbidity or mortality. PMID- 12740137 TI - Determinants of long-term satisfaction after vertical banded gastroplasty. AB - BACKGROUND: The long-term usefulness of vertical banded gastroplasty (VBG) in achieving weight loss is controversial, and adverse effects related to the procedure may attenuate patient satisfaction. Our objective was to evaluate patient satisfaction, and to identify parameters that are related to such satisfaction, 3 to10 years after VBG. METHODS: All consecutive patients who underwent VBG in one surgical ward were invited for a follow-up study 3 to 10 years after surgery. Questions relating to symptoms and quality of life were evaluated in a personal interview using a structured questionnaire. RESULTS: Of the 122 patients who underwent VBG from 1986 to 1992, 75 patients were located and agreed to participate in the follow-up study. The average time since surgery was 5.4 +/- 1.8 years. The average weight loss was 24.9 +/- 12.4%, representing an excess body-weight loss of 58.6 +/- 30.4%. Overall, 65% of the patients were satisfied with the results of surgery while 19% expressed dissatisfaction. Significant improvement was seen in respiratory difficulties, ability to perform physical exercise, and mental status. Successful weight loss and the frequency of respiratory difficulties were the only independent parameters associated with patient satisfaction. Although vomiting, gastroesophageal reflux and difficulty in swallowing occurred in over two-thirds of the patients, their presence was not correlated with patient dissatisfaction. CONCLUSION: Despite the presence of a multitude of adverse effects, the majority of our patients were satisfied with the long-term results of VBG. Successful weight loss and improvement in respiratory difficulties were the major determinants of patient satisfaction. PMID- 12740138 TI - The Lap-Band system in a North American population. AB - BACKGROUND: European and Australian results with laparoscopic adjustable gastric banding (LAGB) using the Lap-Band (Inamed Health, Santa Barbara, CA) have been impressive, with over 100,000 procedures completed at this writing. However, prior to U.S. FDA approval in June 2001, U.S. patients had to travel out of the U.S. for this procedure. This study reports on a series of U.S. patients who requested off-shore referral for LAGB placement. METHODS: 105 U.S. patients were implanted with the Lap-Band System in Mexico by one surgeon in a private practice. 70% were implanted with the perigastric approach, while the final 30% were implanted using the pars flaccida approach. Routine postoperative visits, including band adjustments, were completed in a private U.S. clinic where medical staff performed frequent small adjustments as necessary to optimize results. Data were collected from concurrent and retrospective chart reviews and from telephone interviews. Summary statistics provided for baseline measures included mean +/- standard deviation. Postoperative measures of weight loss included mean +/- standard error. RESULTS: Weight loss results were comparable to international results: 61% EWL at 12 months (n=50), 75% EWL at 24 months (n=37), 72% EWL at 36 months (n=24), and 60% EWL at 48 months (n=7). There were few major complications. CONCLUSION: Attention to patient management is essential to success, and this study found that appropriately-managed U.S. LAGB patients can be as successful as their international counterparts. Frequent follow-up delivered by a bariatric team with easy access to band adjustments is essential. PMID- 12740139 TI - Gastroscopic removal of an adjustable gastric band after partial intragastric migration. AB - BACKGROUND: Laparoscopic implantation of an adjustable gastric band is being performed widely. One potential complication is the transgastric migration of the band, that should be extracted. METHODS: The authors report a series of 182 patients, followed prospectively, October 1996 - April 2002, who had undergone insertion of the Swedish adjustable gastric band. All implantations had been completed by laparoscopy alone. RESULTS: There were no deaths. 15 complications were detected, of which 7 were intragastric migrations of the band (3.8%) at an average follow-up of 40 months. 6 were treated successfully by gastroscopy only, with a new cutter device and without complications. CONCLUSION: The endoscopic technique is beneficial even when the intraluminal migration is partial. PMID- 12740140 TI - Repositioning the Lap-Band for proximal pouch dilatation. AB - BACKGROUND: Laparoscopic adjustable gastric banding (LAGB) procedures have proved their efficiency and reproducibility in several studies. The most frequent late complication is proximal pouch dilatation, with possible progression to total food intolerance. MATERIALS AND METHODS: In a series of 727 laparoscopic bandings using the Lap-Band System, 54 patients presented proximal pouch dilatation and required laparoscopic reposition of the band. 2 patients who had had LAGB placed in another hospital received the same treatment. RESULTS: No particular intra- or postoperative complications occurred during laparoscopic repositioning of the band. 2 conversions were necessary in the beginning of the experience to safely unlock the band. After a median follow-up of 74 months, there has been no recurrence of proximal dilatation. CONCLUSIONS: Laparoscopic repositioning of the Lap-Band System for proximal pouch dilatation is a safe and reproducible procedure which can be proposed as an interesting alternative to its replacement by a new one. An initial perigastric placement of the band allows, during the redo, safe dissection in a virgin pars flaccida tunnel. The calibration of the tiny proximal pouch, the presence of postoperative adhesions, and maintainance of strict control of dietary behavior are probably the reasons for the absence of recurrence of pouch dilatation. PMID- 12740141 TI - Codification of techniques for reoperation after Lap-Band. AB - BACKGROUND: The Lap-Band is a safe, reversible, minimally invasive, and effective bariatric technique, whose main late complications are herniation/dilation of the pouch, erosion/migration of the band, and problems with the adjustment system (subcutaneous access port and connection tubing). We describe the surgical timing and techniques for dealing with the principal complications. METHODS: Between October 1995 and September 2001, we treated 540 patients (mean age 43 years, mean BMI 42.6 kg/m(2)). All operations were completed laparoscopically, with no intra operative complications or mortality. Late complications were: problems with the band adjustment system (n=35); herniation/dilations of the pouch (n=14); erosions/migration of the band into the stomach (n=3). RESULTS: We dealt successfully with all complications using a minimally invasive technique. Herniation/dilations of the pouch necessitated 15 successful repositionings of the band, and 2 removals of the band at the patients' request. Erosion/migration was treated by endo-laparoscopic band removal. Problems with the band adjustment system had to be corrected laparoscopically, under general anesthesia in 17 patients. CONCLUSION: While emphasizing the rarity of complications, we have codified re-operation techniques to help Lap-Band users solve them using a minimally invasive procedure. In particular, problems with the port and connection tubing system can be completely avoided with a transverse subcutaneous positioning in a straight line. PMID- 12740142 TI - Adjustable gastric banding in a multicenter study in Turkey. AB - BACKGROUND: Adjustable gastric banding (AGB) is a minimally-invasive approach which allows adjustment of gastric restriction. METHODS: The AGB was evaluated retrospectively in a consecutive series at 3 centers. From October 1998 to October 2001, 70 patients (49 women), mean age 34.3 years (18-59) with morbid obesity (preoperative mean BMI 45.2 kg/m(2)) underwent AGB The open approach was employed in the first 35 patients. Laparoscopic placement was used in the second 35 patients. Complete follow-up has been obtained in all patients. RESULTS: Mean postoperative follow-up has been 18 months (12-39). Mean operative time was 120 minutes in the open approach and 150 minutes in the laparoscopic AGB. Mean hospital stay was 5 days after the open approach and 1.7 days after the laparoscopic surgery. The excess weight loss after 18 months was 59%. Incidence of early postoperative complications was 27.1%, including nausea and vomiting in 8 patients (5 in open approach, 3 in laparoscopic placement), wound infection in 10 patients (all 10 in open approach), and Wernicke's encephalopathy in 1 patient (open approach). Incidence of late complications was 28.5%, and included band migration in 2 patients (both by laparoscopic placement), pouch dilatation in 10 patients (6 in open approach, 4 in laparoscopic placement), incisional hernias in 4 patients (all by open approach), and port infections in 4 patients (all 4 in open approach). CONCLUSION: AGB has been effective in achieving good weight loss to 3 years follow-up. The ability to adjust the degree of gastric restriction has enabled progressive weight loss. PMID- 12740143 TI - Pressure-induced rhabdomyolysis after bariatric surgery. AB - Rhabdomyolisis most commonly occurs after muscle injury, alcohol ingestion, drug intake and exhaustive exercise. Prolonged muscle compression at the time of surgery may produce this complication. Obesity has been reported as a risk factor for pressure-induced rhabdomyolysis, but no reports associated with bariatric surgery could be found in the literature. We report 3 superobese patients who developed rhabdomyolysis after bariatric surgery. This complication was attributed to direct and prolonged pressure of the bed against the dorsal and gluteal muscles. PMID- 12740144 TI - Rhabdomyolysis of gluteal muscles leading to renal failure: a potentially fatal complication of surgery in the morbidly obese. AB - BACKGROUND: Rhabdomyolysis is a well-known cause of renal failure and is most commonly caused by ischemia/reperfusion or crush injury. We describe a new cause of this syndrome in a series of 6 patients who underwent necrosis of the gluteal muscles after bariatric surgery, 3 of whom eventually died of renal failure. METHODS: Potential etiologic factors were studied by comparing these patients with a consecutive series of 100 patients undergoing primary uncomplicated bariatric surgery during a 1-year period. Demographics, preoperative BMI, co morbidities, duration of operation, and postoperative creatinine phosphokinase (CPK) levels. RESULTS: All patients presented with an area of buttock skin breakdown initially diagnosed as a simple decubitus ulcer. All had extensive myonecrosis of the medial gluteal muscles requiring extensive debridement. 5 of the 6 patients were male, with median BMI 67 compared with a median BMI 55 in the control group (P=0.0022). The patients were on the operating-room table for a median of 5.7 hours compared with 4.0 in the control group (P=0.01). 3 of the 6 developed renal failure requiring dialysis, which was fatal in all. One other patient developed a transient elevation of BUN and creatinine which did not require dialysis. Since recognition of this pattern, we now routinely perform serial CPK measurements. Median CPK rise in uncomplicated patients was to 1,200 mg/dl (SD 450-9,000), while CPK in affected patients ranged from 26,000 to 29,000 IU/l. We now routinely add additional buttock padding in very obese patients and institute aggressive hydration and mannitol diuresis if CPK rises above 5,000. No cases have occurred in the past 18 months in 220 patients. CONCLUSIONS: This is an important and potentially fatal complication of bariatric surgery. Very obese male patients with prolonged surgery are at risk of gluteal muscle necrosis with consequent renal failure, which we hypothesize is due to pressure by the operating-table leading to rhabdomyolysis and the creation of a compartment syndrome. Prevention may be aided by attention to intraoperative padding and positioning, and by limiting the duration of the operation. PMID- 12740145 TI - Lumbar muscle rhabdomyolysis as a cause of acute renal failure after Roux-en-Y gastric bypass. AB - BACKGROUND: Rhabdomyolysis occurs when injury to skeletal muscle disrupts the integrity of the sarcolemmal membrane, allowing release of intracellular proteins into the circulation. Serious complications, such as hyperkalemia, hypocalcemia, hyperphosphatemia, compartment syndrome, cardiac dysrhythmias, disseminated intravascular coagulation, and acute renal failure can develop if diagnosis and treatment are delayed. METHODS: A morbidly obese patient is presented who developed this rare complication after Roux-en-Y gastric bypass. Etiology, pathophysiology, complications, diagnosis and treatment are reviewed, to enable prompt treatment. RESULTS: The patient was treated with crystalloid resuscitation, mannitol, and sodium bicarbonate, and underwent 3 courses of hemodialysis. Normal renal function returned by postoperative day 5. CONCLUSIONS: Morbidly obese patients are at higher risk for developing postoperative rhabdomyolysis, likely because of increased compressive pressure due to the patient's weight. Surgeons should consider rhabdomyolysis in morbidly obese patients who experience postoperative oliguria. Frequent position changes during operations lasting more than 2 hours can protect muscle tissue from compressive injury. PMID- 12740146 TI - Elevated liver enzymes as an operative complication of gastric bypass surgery. AB - BACKGROUND: A rise in liver function tests (LFTs) had been noted in a patient following gastric bypass. This was studied in a subsequent gastric bypass patient, to determine if this rise in LFTs occurred sometime between placement of the upper abdominal retractors but before the retractors were removed. METHODS: LFTs were drawn preoperatively, immediately before retractor placement, immediately before retractor removal, 1 and 3 hours postoperatively, and postoperatively for 5 days until discharge. RESULTS: LFTs rose after liver retractor placement and remained elevated for 3 days before returning to normal. CONCLUSIONS: The authors postulate that surgical retractor placement may have caused the patient's acute LFT elevation. Specifically, direct mechanical pressure may result in a transient decrease in hepatic blood-flow. PMID- 12740148 TI - Similarity of Magenstrasse-and-Mill and Mini-Gastric bypass. PMID- 12740147 TI - Reasonable solutions to weight regain after gastric bypass. PMID- 12740149 TI - A method for treatment of gastric slippage after adjustable gastric banding. PMID- 12740150 TI - Rivastigmine: an update on therapeutic efficacy in Alzheimer's disease and other conditions. AB - The results of recent clinical trials with rivastigmine show that in the approved indication of mild to moderate Alzheimer's disease (AD), the drug is effective in the long term. Rivastigmine produces a significant delay in the decline of the three components of AD that have been identified by European guidelines as essential parameters for the assessment of therapeutic efficacy of medicinal products with this indication. These are cognitive function, the ability to perform the usual activities of daily living, and global judgement of the patient's condition by the patient himself, his caregiver and his doctor. Moreover, rivastigmine produces significant control of AD behavioural disorders. This further reduces caregiver burden, reduces the probability of institutionalisation, and enables the reduction or discontinuation of expensive and poorly tolerated antipsychotics. Recent trials also suggest that rivastigmine is effective in moderate to severe Alzheimer's disease, 'mixed' dementia (AD associated with vascular disorders) and Lewy body dementia. Preliminary investigations have also indicated that the drug may provide important benefits in patients with vascular dementia or dementia associated with Parkinson's disease. Pharmacoeconomic studies show that the therapeutic properties of rivastigmine result in economic savings for the care of demented patients living in the community. PMID- 12740151 TI - Comparison of recombinant human luteinising hormone (r-hLH) and human menopausal gonadotropin (hMG) in assisted reproductive technology. AB - Follicle-stimulating hormone (FSH) and luteinising hormone (LH) act in concert in the stimulation of folliculogenesis and ovulation. However, high levels of LH promote follicular atresia and early miscarriage, and this has led to the concept of a 'therapeutic window' of LH for successful conception in assisted reproductive technology (ART) and ovulation induction. Until now, urinary-derived human menopausal gonadotropin (hMG) has been the only available source of exogenous LH activity. hMG preparations contain highly variable levels of LH, and are often augmented with human chorionic gonadotropin (hCG), which mimics LH activity. Accumulation of hCG bioactivity, however, may have detrimental effects on follicular development and oocyte quality. Recombinant human LH (r-hLH) (Luveris) is the only pure source of LH activity. r-hLH is well characterised and production is tightly controlled, resulting in a highly consistent product. Clinical studies in hypogonadotropic hypogonadal women have demonstrated the efficacy of r-hLH, 75 IU/day, together with r-hFSH, 150 IU/day, in promoting optimal follicular development, oestrogen secretion and endometrial thickness. r hLH therefore provides the clinician with the opportunity for precise and consistent dosing within the therapeutic window for patients requiring exogenous LH, without the risk of LH overexposure that is associated with hCG. PMID- 12740152 TI - Experience with COSOPT, the fixed combination of timolol and dorzolamide, gained in Swiss ophthalmologists' offices. AB - PURPOSE: In managing glaucoma through the lowering of intraocular pressure (IOP), beta-blockers (e.g. timolol) are commonly the drug of first choice, with other drugs added to control IOP such as carbonic anhydrase inhibitors (e.g. dorzolamide). The present survey was instigated to assess the COSOPT (fixed combination of topical timolol and topical dorzolamide) prescription behaviour of Swiss office-based ophthalmologists and to evaluate the IOP-lowering efficacy of COSOPT under real-life private practice conditions. METHODS: A survey was conducted in Swiss ophthalmologist practices. Ophthalmologists were asked to report their experience with COSOPT therapy prescribed to patients with glaucoma or ocular hypertension. Parameters recorded were: patient demographics, IOP, reason for choosing COSOPT, continuation of COSOPT therapy after completion of the survey and, if stopped, reason for discontinuation, and spontaneously occurring side-effects. All entries were optional. Since the purpose of this survey was to obtain a realistic picture of daily practice in Swiss ophthalmologists' offices, no guidelines with regard to measurement of IOP, time between visits and actual treatment were given. RESULTS: Data of 733 patients were included in this analysis. COSOPT as sole therapy was prescribed to 538 patients (74%), whereas 120 (16%) were put on a combination regimen of COSOPT + latanoprost. The remaining 75 patients (10%) received combinations of COSOPT with other ocular hypotensives. When assessing those patients put on COSOPT alone, IOP reduction in new, previously untreated patients was 10.8 mmHg (average), 5.4 mmHg (average) in upgrades from previous monotherapy and 2.7 mmHg after switch from free combination therapy. Patients treated with a combination of COSOPT + latanoprost showed an IOP reduction of 27 mmHg (new, previously untreated patients), 7.6 mmHg (upgrades from monotherapy) and 3.8 mmHg (switches from previous free combination regimen). Therapy was continued in 89% of all patients overall and in 91% of new patients treated with COSOPT alone (first line). 6.8% stopped therapy due to side-effects. CONCLUSIONS: COSOPT, the fixed combination of 0.5% timolol and 2% dorzolamide, is broadly prescribed to new, upgrade and switch patients in Switzerland. COSOPT therapy is well-tolerated and highly efficacious for patients requiring strong IOP lowering. Nine out ten patients stay on therapy after 1-2 months of treatment. PMID- 12740153 TI - Higher glutathione transferase GSTM1 0/0 genotype frequency in young thyroid carcinoma patients. AB - OBJECTIVE: Glutathione S-transferases (GSTs) comprise a large supergene family and detoxify a variety of endogenous and exogenous electrophilic compounds. Since many GSTs are polymorphic, there has been considerable interest in determining whether particular allelic variants are associated with altered risk for various disorders. RESEARCH DESIGN AND METHODS: In this study the association between the variant GSTM1 0/0 genotype and thyroid carcinoma was investigated. A hospital based, case-controlled study was carried out. Polymorphisms of GSTM1 0/0 (i.e. the null allele of GSTM1) in samples from 32 cases and 44 controls were detected by polymerase chain reaction (PCR) methodology. The proportions of GSTM1 deleted genotype in cases and controls were 59.4% and 54.5%, respectively. RESULTS: There were significant increments of GSTM 0/0 genotype frequency in a group of patients aged under 40 (p = 0.033, odds ratio (OR) = 4.78, 95% confidence interval (CI) = 1.30-7.13) and in former smokers compared with controls (p = 0.039, OR 2.45, 95% CI 0.216-4.72). CONCLUSION: GSTM1 deleted genotype may be a useful genetic biomarker for thyroid carcinoma susceptibility in young subjects. The absence of this enzyme seems to have a role in the development of thyroid carcinoma; however, the mechanism still needs further study. PMID- 12740154 TI - Tolerability of percutaneous coronary interventions in patients receiving nadroparin calcium for unstable angina or non-Q-wave myocardial infarction: the Angiofrax study. AB - BACKGROUND: Nadroparin, a low-molecular-weight heparin (LMWH), is an alternative to unfractionated heparin for the acute management of patients with non-ST elevation acute coronary syndrome (ACS): unstable angina or non-Q-wave myocardial infarction. However, unfractionated heparin can be substituted for LMWH in patients requiring percutaneous coronary interventions (PCIs) for the duration of the procedure. The tolerability of this anti-thrombotic regimen (i.e. unfractionated heparin for the duration of PCIs, preceded and followed by subcutaneous injection of nadroparin) is not yet documented. DESIGN AND METHODS: This open-label 6-day study was carried out in 302 patients to test the tolerability of this anti-thrombotic regimen in patients requiring PCIs. The primary end-point of the study was the incidence of major haemorrhage over the whole study duration (6 days). The secondary end-point was the need for transfusion and vascular repair after PCI. RESULTS: The incidence of major haemorrhage in patients undergoing coronary angiography (CA) without or with PCIs was 1.4% and 1.3%, respectively, and the incidence of minor haemorrhage was 10.7% and 23.5%, respectively. These results are consistent with published data. CONCLUSIONS: These results suggest that CA and PCIs can be performed safely in patients being treated for unstable angina or non-Q-wave myocardial infarction receiving nadroparin pre- and post-coronary procedure and/or intervention, substituted by unfractionated heparin for the duration of the intervention. PMID- 12740155 TI - The ELIXIR study: evaluation of sexual dysfunction in 4557 depressed patients in France. AB - OBJECTIVES: This survey was conducted in order to determine the extent and nature of disorders of sexual function in depressed patients treated in the community in France. METHODS: Patients with DSM-IV major depressive episodes were included. The inclusion criteria stipulated that only patients with no antecedents of sexual dysfunction could be included. Information on sexual function was collected with a questionnaire which included physician observations as well as the Arizona Sexual Experience Scale. RESULTS: Overall, 4557 patients were included in the study. The prevalence of disorders of sexual function observed was 35% for spontaneously reported problems and 69% for problems identified by physician questioning. Impaired sexual function in depressed patients is also revealed by a high score on the Arizona Sexual Experience Scale (mean overall score of 21.4). Frequency of sexual dysfunction was somewhat higher in patients treated with antidepressants than in untreated patients (71% and 65% respectively). Treatment with tianeptine was associated with a lower incidence of sexual dysfunction than was treatment with tricyclic antidepressants or with selective serotonin reuptake inhibitors. Although in 39% of cases, physicians managed the sexual problems encountered by changing the antidepressant treatment, the most frequently adopted approach (42% of cases) was to await spontaneous remission. Drug holidays or adjunctive therapy were very rarely proposed. CONCLUSIONS: The prevalence of sexual dysfunction in patients with major depression is high. Antidepressant drugs appear to aggravate such problems, with certain classes of drug better tolerated than others. Sexual dysfunction in depressed patients is often not optimally treated. PMID- 12740156 TI - Repetitive transcranial magnetic stimulation (rTMS): new tool, new therapy and new hope for ADHD. AB - Attention-deficit hyperactivity disorder (ADHD) is the most common developmental disorder that is associated with environmental and genetic factors. Neurobiological evidence suggests that fronto-striatum-cerebellum circuit abnormalities, mainly in the right hemisphere, are responsible for most of the disturbed sensorimotor integration; dopamine seems to be the main neurochemical alteration underlying these morphological abnormalities. Different conventional treatments have been employed on ADHD; however, repetitive transcranial magnetic stimulation (rTMS), a new and useful option for the clinical/research investigation of several neuropsychiatric disorders involving dopamine circuits, has yet to be considered as a therapeutic tool and possible drug-free option for ADHD. Here the authors explore the available evidence that makes this tool a rational therapeutic possibility for patients with ADHD, calling attention to safety issues, while highlighting the potentials of such an approach and the new hope it may bring for patients, parents, researchers and clinicians. The authors advocate carefully conducted clinical trials to investigate efficacy, safety, cost-effectiveness and clinical utility of rTMS for ADHD patients - in comparison to both placebo and standard treatments. PMID- 12740157 TI - Cyclo-oxygenase-2 (COX-2) mRNA expression correlates with progesterone receptor positivity in human breast cancer. AB - BACKGROUND: We previously presented evidence showing that cyclo-oxygenase 2 (COX 2) plays an important role in mammary carcinogenesis and angiogenesis in human breast cancer. The present study aims to compare COX-2 mRNA expression with hormone receptor status, S-phase fraction, telomerase activity, and DNA ploidy in human breast cancer. METHODS: Total cellular RNA was extracted from frozen breast tissue samples according to standard methodology. The mRNA copy numbers for COX-2 were determined in 18 infiltrating carcinomas using quantitative RT-PCR and TaqMan methodology. The oestrogen receptor (ER) and progesterone receptor (PgR) status was determined using the ligand-binding technique (ER+ = > 3 fmol/mg, PgR+ = > 5 fmol/mg). We also determined DNA ploidy status (diploid or aneuploid), S phase fraction (< 6% = low, 6-10% = intermediate, > 10% = high), and telomerase activity (total protein generated by TRAP assay). RESULTS: The median COX-2 mRNA copy number per micro g of RNA was 126 713 (range = 15 717-2 022 050). COX-2 expression was significantly associated with PgR positivity (p = 0.013). The association between COX-2 and DNA diploidy failed to reach a statistical significance (p = 0.085). No significant association was detected between COX-2 and S-phase fraction, ER status, or telomerase activity. CONCLUSIONS: COX-2 mRNA expression is associated with PgR positivity in human breast cancer. This observation is consistent with the hypothesis that COX-2 upregulates aromatase activity. PMID- 12740158 TI - Efficacy and safety of stabilised hydrogen peroxide cream (Crystacide) in mild-to moderate acne vulgaris: a randomised, controlled trial versus benzoyl peroxide gel. AB - BACKGROUND: Benzoyl peroxide (BP) is a first-line topical treatment in acne vulgaris (AV). However, its use can cause mild skin irritation and dryness. A new formulation of hydrogen peroxide stabilised (HPS) in monoglycerides cream (Crystacide 1%), indicated in the topical treatment of superficial skin infections, is now available as an alternative treatment. STUDY AIM: To evaluate efficacy and local tolerability of HPS in mild-to-moderate AV in comparison with BP gel. METHODS AND PATIENTS: In a randomised, prospective, investigator-masked parallel-group, 8-week trial, 60 patients (24 men, 36 women, mean age 25 +/- 6 years) with mild-to-moderate AV, affecting mainly the face, were enrolled in the study, after their informed consent. HPS or BP (PanOxyl gel 4%) was applied topically twice daily for 8 weeks. STUDY OUTCOMES: The study endpoints were: (1) Reduction in mean inflammatory (IL), noninflammatory (NIL) and total (TL) acneic lesions in comparison with baseline; (2) Local tolerability assessed evaluating erythema, dryness and burning sensation, using a 0-3 qualitative score (score 0 = poor tolerability; score 3 = very good tolerability). RESULTS: TL, NIL, and IL were assessed by an investigator unaware of treatment allocation at baseline, and week 8. The tolerability score (TS) was assessed at week 4 and 8. At baseline, the two groups were well matched for the main clinical and demographic characteristics. All patients concluded the trial. At week 0, in the HPS group TL, NIL and IL (mean +/- SD) were: 35 +/- 8, 20 +/- 6 and 16 +/- 7. At week 8, HPS reduced TL to 16 +/- 7; NIL to 9 +/- 3 and IL to 7 +/- 3 (p < 0.001). At baseline, TL, NIL and IL, in the BP group, were 32 +/- 9, 24 +/- 8 and 18 +/- 7, respectively. At week 8, BP reduced TL, NIL and IL to 14 +/- 9; 7 +/- 5 and 7 +/- 3 (p < 0.001). In comparison with baseline values, the percentage reductions of IL were 58% and 61% for HPS and BP,respectively (p = n.s.). At the end of the study the TS was 2.9 +/- 0.2 in HPS group and 2.4 +/- 0.8 in BP group (p < 0.025). Two patients in HPS group (6%) and seven patients (23%) in BP group suffered from mild-to-moderate local erythema. CONCLUSIONS: HPS has shown to be as effective as BP in reducing both inflammatory and noninflammatory AV lesions in patients with mild-to-moderate disease. In comparison with BP 4% gel, HPS cream shows a better local tolerability profile. PMID- 12740159 TI - Clinical pharmacokinetics of fenofibrate. PMID- 12740161 TI - The home office and the dangerous trades. Regulating occupational disease in Victorian and Edwardian Britain. PMID- 12740170 TI - High-concentration tramadol-induced vasodilation in rabbit aorta is mediated by both endothelium-dependent and -independent mechanisms. AB - AIM: The mechanism of tramadol-induced vasodilation was investigated using isolated rabbit thoracic aortic rings. METHODS: Aortic rings from 8 rabbits were placed in organ bath and precontracted with phenylephrine (10(-5) mol/L) before addition of tramadol. Relaxation responses by tramadol were evaluated in the presence and absence of endothelium, indomethacin (an inhibitor of cyclooxygenase), NG-nitro-L-arginine methyl ester (L-NAME, a specific inhibitor of nitric oxide synthase), glibenclamide (an inhibitor of ATP-sensitive potassium channels), tetraethylammonium chloride (TEA, an inhibitor of calcium-sensitive potassium channels), and naloxone (an antagonist of opioid receptors). RESULTS: Tramadol (10(-4) mol/L and 3 x 10(-4) mol/L) caused significant vasodilation in endothelium-intact and endothelium-denuded aortic rings (P<0.05). The relaxation response to tramadol was significantly greater in endothelium-intact rings than in endothelium-denuded rings. Pretreatment of aortic rings with indomethacin (10( 5) mol/L), glibenclamide (10(-5) mol/L), TEA (10(-3) mol/L), and naloxone (10(-4) mol/L) had no effect on the tramadol-induced relaxation. In endothelium-intact rings, L-NAME (10(-4) mol/L) pretreatment caused marked inhibition of the relaxation induced by tramadol, but not endothelium-denuded rings. CONCLUSION: In the rabbit aorta, vascular relaxation induced by tramadol is due to both nitric oxide production from endothelium and a direct effect on smooth muscle. PMID- 12740171 TI - EDT, a tetrahydroacridine derivative inhibits cerebral ischemia and protects rat cortical neurons against glutamate-induced cytotoxicity. AB - AIM: To study the effects of 9-(4-ethoxycarbonylyphenoxy)-6,7-dimethoxy-1,2,3,4 tetrahydroacridine (EDT) on cerebral ischemia and glutamic acid (Glu) and sodium nitroprusside (SNP)-induced neurocytotoxicity in primary cortical culture. METHODS: Focal cerebral ischemia was produced by permanent occlusion of left middle cerebral artery (MCA) in mice. The infarct tissue was measured by 2,3,5 triphenyltetrazolium chloride (TTC) staining technique. The extent of neurological deficits was evaluated. In primary cortical culture, colorimetric MTT assay was used to determine cell survival rate, and leakage of LDH and NO release assay were measured. RESULTS: In focal cerebral ischemia, pretreatment with EDT 2.5, 5, and 10 mg/kg and nimodipine 2 mg/kg for 5 d effectively improved the abnormal neurological symptoms and reduced the infarct rate. In primary cortical culture, EDT 0.01-3 micromol/L concentration-dependently attenuated NO release induced by Glu 500 micromol/L and increased the cell survival. It also remarkably reduced the LDH excessive efflux. CONCLUSION: EDT possessed protective effects against cerebral ischemia, which may be related to blocking Glu receptor and inhibiting NO formation. PMID- 12740172 TI - Effect of antisense oligonucleotide of noggin on spatial learning and memory of rats. AB - AIM: To investigate the effect of antisense oligonucleotide (ASODN) of noggin on rat spatial learning and memory. METHODS: Expression of noggin mRNA was measured by in situ hybridization method and the ability to spatial learning and memory was tested with Morris water maze. RESULTS: Compared with control rats, noggin mRNA positive neurons in dentate gyrus (DG) and CA3 region of hippocampus were markedly increased after the Morris water maze training (P<0.01). The increase of noggin mRNA positive neurons in hippocampus following maze training could be significantly blocked by icv injection of antisense noggin ODN, and the injection also impaired the learning and memory formation as compared to that in control rats. But the sense oligonucleotide (SODN) had no effect. CONCLUSION: Noggin, as an embryonic gene expressed in adult hippocampus, plays an important role in the process of learning and memory formation. PMID- 12740173 TI - Different synaptic mechanisms of long-term potentiation induced by nicotine and tetanic stimulation in hippocampal CA1 region of rats. AB - AIM: To investigate whether long-term potentiation (LTP) induced by nicotine and tetanic stimulation in the hippocampal CA1 region shares different mechanisms. METHODS: Extracellular population spikes of the pyramidal cell layer in the hippocampal CA1 region were recorded in vitro. RESULTS: LTP induced by the tetanic stimulation could be facilitated by nicotine 10 micromol/L, meanwhile, the tetanic stimulation did the same effect on LTP induced by nicotine 10 micromol/L. MK-801 10 micromol/L or NG-nitro-L-arginine methyl ester (L-NAME) 20 micromol/L blocked LTP induced by tetanic stimulation, but did not inhibit LTP induced by nicotine. CONCLUSION: LTP induced by nicotine shares different synaptic mechanisms with LTP induced by tetanic stimulation. PMID- 12740174 TI - Effects of phenytoin on morphology and structure of hippocampal CA3 pyramidal neurons of rats in chronic stress. AB - AIM: To investigate the effects of phenytoin (DPH) on morphological and structural changes of pyramidal neurons in hippocampal CA3 of rats induced by chronic stress. METHODS: Using Nissl staining, Golgi staining, and electron microscope, the morphology and structure of pyramidal neurons in hippocampal CA3 of rats were observed. RESULTS: Chronic stress resulted in loss of hippocampal CA3 pyramidal neuron from 39+/-4 to 35+/-4, shortening of total length of apical dendrite (from 196 microm+/-35 microm to 156 microm+/-33 microm, P<0.05), and ultrastructural degenerations of neurons. DPH markedly inhibited the decreases in number of hippocampal CA3 pyramidal neuron (38.4+/-2.2) and total length of apical dendrite (198 microm+/-36 microm, P<0.05), meanwhile, improved neuron ultrastructural degenerations caused by chronic stress. CONCLUSION: Chronic stress does damage to hippocampal CA3 pyramidal neurons and DPH protects hippocampus from damage induced by chronic stress. PMID- 12740175 TI - Isoprenaline and aminophylline relax bronchial smooth muscle by cAMP-induced stimulation of large-conductance Ca2+-activated K+ channels. AB - AIM: To investigate whether the relaxation of bronchial smooth muscle induced by isoprenaline and aminophylline is mediated by large conductance Ca2+-activated K+ channels (BK(Ca)) via cAMP-dependent mechanism. METHODS: With isometric tension recording, the role of BK(Ca) in relaxations of rat bronchial strips induced by isoprenaline and aminophylline was determined. With perforated patch-clamp technique, BK(Ca) currents were observed in freshly isolated rat bronchial myocytes. RESULTS: Tetraethylammonium 5 mmol/L, a BK(Ca) blocker, caused a significant rightward shift in the concentration-response curves of isoprenaline and aminophylline (about 4.26-fold and 3.78-fold, respectively) in methacholine precontracted rat bronchial strips. Isoprenaline 1 micromol/L caused a significant increase in BK(Ca) current from (94+/-15) pA/pF to (186+/-30) pA/pF (voltage steps from -60 mV to +50 mV, n=10, P<0.01), which was partly abolished by Rp-cAMP 100 micromol/L, a protein kinase A inhibitor. Furthermore, current voltage relationship(I-V) curve exhibited an upward shift, and the peak current density was significantly raised (n=10, P<0.01) by ramp depolarization from -100 mV to +100 mV. Aminophylline 1 mmol/L caused a significant increase in BK(Ca) current from (90+/-10) pA/pF to (166+/-25) pA/pF (voltage steps from -60 mV to +50 mV, n=11, P<0.01), which was partly abolished by Rp-cAMP 100 micromol/L. Furthermore, the I-V curve exhibited an upward shift, and the peak current density was significantly raised (n=11, P<0.01) by ramp depolarization from -100 mV to +100 mV. CONCLUSION: The relaxations induced by isoprenaline and aminophylline were, at least partly, mediated by cAMP-stimulation of BK(Ca) in rat bronchial smooth muscle. PMID- 12740176 TI - Actinomycin D inhibiting K562 cell apoptosis elicited by salvicine but not decreasing its cytotoxicity. AB - AIM: To study the effects of actinomycin D (Act D) on the cytotoxicity and apoptosis elicited by salvicine in human leukemia K562 cells. METHODS: Growth inhibition of K562 cells was measured by the microculture tetrozolium (MTT) assay. Cell apoptosis was evaluated by fluorescence microscopy, DNA agarose gel electrophoresis, and flow cytometry. RESULTS: Following exposure of K-562 cells to salvicine plus Act D for 24 h, Act D at the concentrations of 0.04, 0.4, and 4 micromol/L potentiated the cytotoxicity of salvicine 6.25 micromol/L to some degree. The mean growth inhibitory rates went from 8 % up to 69 %, 71 %, and 70 %, respectively. However, the same enhancement of Act D did not continue to emerge at the higher concentrations than salvicine 6.25 micromol/L. Act D enhanced, or at least, did not decrease the cytotoxicity of salvicine against K562 cells. Fluorescence microscopy, DNA agarose gel electrophoresis, and flow cytometry revealed that Act D concentration-dependently inhibited the induction of apoptosis by salvicine in the same cell line. CONCLUSION: The combination of salvicine and Act D in a proper range of concentrations is able to enhance the cytotoxicity of salvicine against K562 cells though inhibiting apoptosis. The other mechanisms of cell death except apoptosis may be implicated in the process. PMID- 12740177 TI - Mechanism underlying enhanced endothelium-dependent vasodilatation in thoracic aorta of early stage streptozotocin-induced diabetic mice. AB - AIM: To investigate the mechanism of the enhanced endothelium-dependent vasodilatation in thoracic aorta of the early stage streptozotocin (STZ)-induced diabetic C57BL/6J mice. METHODS: Radioimmunity was used to detect the metabolite of prostaglandin I2 (PGI2), 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha), in the blood serum. Vascular muscle tension and phenylephrine (PE)-induced rhythmic activity in the isolated thoracic aorta of mice were also compared. RESULTS: 6 Keto-PGF1 alpha in the serum was significantly higher in STZ-induced diabetic mice than age-matched controls [(1.8+/-1.0) microg./L vs (0.5+/-0.3) microg/L, P<0.01]. PE induced rhythmic activity in both diabetic and control mouse aorta but the amplitude was markedly higher in diabetic mice than in controls [(4.9+/ 1.7) % vs (12+/-5) %, P<0.01]. PE, high K+ solution-induced contraction, and acetylcholine (ACh)-induced relaxation [(56+/-10) % vs (81+/-8) %, P<0.01] were notably enhanced in diabetic mice than those in controls. Alone NG-nitro-L arginine methyl ester (L-NAME) or 6-(phenylamino)-5,8-quinolinedione (LY-83583) abolished the rhythmic activity and ACh-induced relaxation in controls but only partially inhibited them in diabetic mice. Indomethacin did not affect rhythmic activity but depressed ACh-induced relaxation. L-NAME plus indomethacin significantly depressed the rhythmic activity and ACh-induced relaxation than L NAME alone (P<0.01). Furthermore tetraethylammonium plus L-NAME abolished them in diabetic mice. CONCLUSION: The mechanism that enhanced endothelium-dependent vasodilatation in STZ-induced diabetic mice is due to enhanced production of PGI2 and endothelium-derived hyperpolarizing factor (EDHF). The phenomena maybe only take place in early stage of diabetic mice. PMID- 12740178 TI - Rosiglitazone reverses insulin secretion altered by chronic exposure to free fatty acid via IRS-2-associated phosphatidylinositol 3-kinase pathway. AB - AIM: To study the effect of rosiglitazone (RSG) on insulin secretion in isolated pancreatic islets under chronic exposure to free fatty acid (FFA) and to investigate the potential signaling mechanism of RSG action. METHODS: Rat pancreatic islets were cultured with or without FFA (2 mmol/L, oleate:palmitate, 2:1) in the presence or absence of RSG (0.05-10 micromol/L). The insulin release was measured by radioimmuoassay, the expression level of insulin receptor substrate-2 (IRS-2) protein and the association of IRS-2 with p85 subunit of phosphatidylinositol 3-kinase (PI 3-kinase) were determined by immunoprecipitation and Western blot. RESULTS: The islets exposed to high FFA concentration showed an increased basal and a decreased glucose-induced insulin release as compared with control islets (P<0.01). IRS-2 protein level was decreased by 65 % (P<0.01) and the association of IRS-2 with p85 subunit of PI 3 kinase and was decreased by 73 % (P<0.01). When islets were cultured with FFA in the presence of RSG 5 micromol/L, both basal and glucose-induced insulin secretion were reversed to a pattern of control islets (P<0.01, P<0.05). The addition of RSG in the cultured medium increased significantly the expression of IRS-2 protein by 2.6 fold (P<0.01) and the association of IRS-2 with p85 by 2.7 fold (P<0.01) as compared with islets incubated with FFA alone. The effects of RSG on insulin secretion were blocked by a PI 3-kinase inhibitor, wortmannin. CONCLUSION: The effects of RSG on insulin secretion could be mediated through an IRS-2-associated PI 3-kinase signaling pathway. PMID- 12740179 TI - Regulatory effect of IFN-gamma on expression of TGF-beta 1, T beta R-II, and StAR in corpus luteum of pregnant rhesus monkey. AB - AIM: To examine expression of transforming growth factor-beta 1 (TGF-beta 1) and TGF beta receptor II (T beta R-II) and steroidogenic acute regulatory protein (StAR) in the corpus luteum (CL) of pregnant monkeys at various stages and to study possible effect of IFN-gamma on their production. METHODS: In situ hybridization and immunohistochemistry were applied to detect mRNA and protein. RESULTS: The expression of StAR, TGF-beta 1, and T beta R-II in the pregnant monkey CL was progressively decreased from d 15 to d 35 of gestation. IFN-gamma down-regulated the expression of TGF-beta 1, T beta R-II, as well as StAR. CONCLUSION: TGF-beta 1 may play an important role in the CL formation and functional maintaining; IFN-gamma down-regulates the expression of TGF-beta 1, T beta R-II, and StAR. PMID- 12740180 TI - Microbial transformation of naproxen by Cunninghamella species. AB - AIM: The metabolites of naproxen produced by Cunninghamella species were isolated and identified, and further to compare the similarities between microbial transformation and mammalian metabolism. METHODS: Naproxen was transformed by three strains of Cunninghammella species (Cunninghamella blakeslesna AS 3.153, Cunninghamella echinulata AS 3.2004, and Cunninghamella elegans AS 3.156). The metabolites of naproxen were separated and assayed by liquid chromatography-mass spectrometry method. Semi-preparative HPLC was used to isolate the major metabolite, and the structure was identified by nuclear magnetic resonance (NMR) and mass spectrometry. RESULTS: Naproxen was transformed into 2 metabolites, desmethylnaproxen and desmethylnaproxen-6-O-sulfate, both were the known mammalian metabolites. The conjugated metabolite was newly detected in microbial transformation samples. CONCLUSION: The microbial transformation of naproxen has some similarities with the metabolism of naproxen in mammals. The fungi belonging to Cunninghamella species could be used as complementary in vitro models for drug metabolism to predict and produce the metabolites of drugs in mammals. PMID- 12740181 TI - Evaluation on drug dependence of buprenorphine. AB - AIM: To survey and assess the drug dependence and abuse potential liability of buprenorphine among opiate abusers. METHODS: Subjects of opiate dependence with history of buprenorphine use for 3 d at least were surveyed by interview. Physical dependence of buprenorphine was assessed using 30 items opiate withdrawal scale (OWS), which composed of 30 symptoms/signs. A 4-point scale was used to rate each symptoms/signs: zero (0), mild (1), moderate (2), and severe (3). Subjects were asked to rate their symptoms according to severity of previous experienced buprenorphine withdrawal. The estimate of the degree of subjective euphoria for buprenorphine was assessed using visual analogue scale (VAS). RESULTS: Subjects 1235 who met the research criteria cases completed this survey in multi-detoxification treatment centers. The main initial purposes of buprenorphine use were detoxification (77.4 %) and protracted abstinence treated (26.6 %) respectively. The scores of OWS of buprenorphine were between 0.2 to 1.3; The mean scores of OWS in 3 different categories of frequency of buprenorphine use on "continuous use", "un-continuous use", and "sometimes continuous, sometimes un-continuous" were 0.9+/-0.9, 0.4+/-0.5, and 0.7+/-0.4, respectively (F=70.846, P<0.05). The degree of subjective euphoria for buprenorphine was slight to sub-moderate (mean score of VAS was 27 mm+/-24 mm). The mean scores of VAS in different routes of buprenorphine administration of sublingual and injection were (24+/-23) mm and (27+/-24) mm, respectively. No significant difference was found between sublingual and injection use of buprenorphine (u=1.516, P>0.05). CONCLUSION: Both physical and psychic dependence of buprenorphine were low. PMID- 12740182 TI - Reliability of phototoxic tests of fluoroquinolones in vitro. AB - AIM: To make sure the reliability of phototoxic tests in vitro by comparing the phototoxic potential of 4 fluoroquinolones (FQ). METHODS: Lomefloxacin (LFLX), sparfloxacin (SPFX), ciprofloxacin (CPFX), and norfloxacin (NFLX) were tested by Wistar rat phototoxic test and Balb/c mouse phototoxic test in vivo, and Chinese hamster V79 cell micronucleus test and NIH 3T3 MTT test in vitro under the different condition of UVA irradiation. RESULTS: In all experiments, LFLX and SPFX showed higher phototoxic potential compared with the control (P<0.01 vs 1 % CMC), CPFX was mild (P<0.05 vs physiological saline), NFLX did not show phototoxicity in vivo, however at a higher concentration (10 micromol/L) in vitro, it also induced phototoxicity as other FQ. CONCLUSION: There are good correlations between phototoxic tests in vivo and in vitro. These results ensure the validation of phototoxic tests in vitro. PMID- 12740183 TI - Molecular simulation of a single-chain antibody against AChE to explore molecular basis of inhibitory effect of 3F3 McAb on enzyme activity. AB - AIM: To explore the molecular basis of the inhibitory effect of 3F3, a monoclonal antibody against acetylcholinesterase (AChE), by computer-aided molecular simulation. METHODS: The single-chain 3F3 antibody (Sc3F3) was designed by joining VH and VL via a flexible linker (Gly4Ser)3. The amino acid sequence of the recombinant Sc3F3 was then subjected to computer-aided molecular modeling, and docking with the antigen molecule AChE to mimic the immunoactive interaction in a three-dimensional fashion. RESULTS: The modeled structure of Sc3F3 manifested the common features of a classical antibody. Both VH and VL were composed of two ?-sheets and connecting loops. The docking profile of the action between Sc3F3 with AChE demonstrated the formation of a stable structure. The van der Waals force played an important role suggesting that the complex was formed mainly via hydrophobic interactions between Sc3F3 and AChE molecules. CONCLUSION: The spatial structure of the complex of Sc3F3 and AChE showed that Sc3F3 overlaid the entrance of the active center gorge of AChE blocking the access of substrate. PMID- 12740184 TI - Effects of Ginkgo biloba extract on acute cerebral ischemia in rats analyzed by magnetic resonance spectroscopy. AB - AIM: To study the effect of Ginkgo biloba extract (GbE) on acute cerebral ischemia in rats. METHODS: The rats were randomly divided into four groups: sham operated group (group I as control), ischemic group (group II), the prophylactic (GbE premedication) group (group III) and GbE-treatment group (group IV). Magnetic resonance spectroscopy (MRS) was carried out to dynamically monitor the changes in biochemical metabolic variations 48 h after cerebral ischemia and effects of GbE (100 mg/kg, ip, qd). RESULTS: (1) Lactate (Lac) peak could be detectable at the infarction area 90 min after acute cerebral ischemia and increased with time. Lac peak in the prophylactic group was elevated slightly (P<0.01, n=6), whereas in the treatment group the elevation of Lac was more remarkable than that in the prophylactic group (P<0.05, n=6). (2) In the ischemic group, the level of N-acetyl aspartate (NAA) was decreased within 4 h after ischemia (P<0.05, n=6), and the decline persisted (P<0.01, n=6). In the treatment group and prophylactic group, NAA was decreased slightly after 24 h (P<0.05, n=6). (3)Twenty-four hours after ischemia, in both ischemic group and treatment group, choline (Cho) was elevated slightly (P<0.05, n=6) and creatine (Cr) was decreased slightly (P<0.05, n=6), but in the prophylactic group these changes occurred only after 48 h. CONCLUSION: GbE could prevent and treat acute cerebral ischemia. The effectiveness was more satisfactory when GbE was used preventively. PMID- 12740185 TI - Hyper-polyhedron model applied to molecular screening of guanidines as Na/H exchange inhibitors. AB - AIM: To investigate structure-activity relationships of N-(3-Oxo-3,4-dihydro-2H benzo[1,4]oxazine-6-carbonyl) guanidines in Na/H exchange inhibitory activities and probe into a new method of the computer-aided molecular screening. METHODS: The hyper-polyhedron model (HPM) was proposed in our lab. RESULTS: The samples with probably higher activities could be determined in such a way that their representing points should be in the hyper-polyhedron region where all known samples with high activities were distributed. And the predictive ability of different methods available was tested by the cross-validation experiment. CONCLUSION: The accurate rate of molecular screening of N-(3-Oxo-3,4-dihydro-2H benzo[1,4]oxazine-6-carbonyl) guanidines by HPM was much higher than that obtained by PCA (principal component analysis) and Fisher methods for the data set available here. Therefore, HPM could be used as a powerful tool for screening new compounds with probably higher activities. PMID- 12740186 TI - Evidence-based information and experience, the perfect match. PMID- 12740187 TI - Practice guidelines for evaluation of Fever in returning travelers and migrants. AB - BACKGROUND: Fever upon return from tropical or subtropical regions can be caused by diseases that are rapidly fatal if left untreated. The differential diagnosis is wide. Physicians often lack the necessary knowledge to appropriately take care of such patients. OBJECTIVE: To develop practice guidelines for the initial evaluation of patients presenting with fever upon return from a tropical or subtropical country in order to reduce delays and potential fatal outcomes and to improve knowledge of physicians. TARGET AUDIENCE: Medical personnel, usually physicians, who see the returning patients, primarily in an ambulatory setting or in an emergency department of a hospital and specialists in internal medicine, infectious diseases, and travel medicine. METHOD: A systematic review of the literature--mainly extracted from the National Library of Medicine database--was performed between May 2000 and April 2001, using the keywords fever and/or travel and/or migrant and/or guidelines. Eventually, 250 articles were reviewed. The relevant elements of evidence were used in combination with expert knowledge to construct an algorithm with arborescence flagging the level of specialization required to deal with each situation. The proposed diagnoses and treatment plans are restricted to tropical or subtropical diseases (nonautochthonous diseases). The decision chart is accompanied with a detailed document that provides for each level of the tree the degree of evidence and the grade of recommendation as well as the key points of debate. PARTICIPANTS AND CONSENSUS PROCESS: Besides the 4 authors (2 specialists in travel/tropical medicine, 1 clinical epidemiologist, and 1 resident physician), a panel of 11 European physicians with different levels of expertise on travel medicine reviewed the guidelines. Thereafter, each point of the proposed recommendations was discussed with 15 experts in travel/tropical medicine from various continents. A final version was produced and submitted for evaluation to all participants. CONCLUSION: Although the quality of evidence was limited by the paucity of clinical studies, these guidelines established with the support of a large and highly experienced panel should help physicians to deal with patients coming back from the Tropics with fever. PMID- 12740188 TI - The effect of education on cognitive performances and its implication for the constitution of the cognitive reserve. AB - Some studies have suggested that people with a high educational level have a lower risk of developing dementia compared to people with a low educational level. This protective effect of education has been explained by the constitution of a cognitive reserve which might delay the cognitive and functional expression of neurodegenerative illnesses. The aim of this study is, on the one hand, to evaluate the impact of education on cognitive functioning, which is thought to support the cognitive reserve capacity, and on the other, to determine the extent to which cognitive functioning is affected by other explanatory variables. The analysis was conducted on 1022 individuals without physical or neurological disorders in the Personnes Ages Quid study. These participants were aged 66 and over and had completed a neuropsychological battery. The effect of some demographic and socioeconomic variables on cognitive performance was also analyzed. Multivariate analysis showed a significant effect of education on most neuropsychological performances, independently of the other variables, and more particularly, in the high-attention-demanding tests. A principal component analysis demonstrated that education specifically increases 2 cognitive components: controlled processes and conceptualization ability. More-over, mental stimulation occurring after the education years, such as high-complex-activity occupations, seems to increase the controlled component. All these results suggest that the effect of education on cognitive reserve may be explained by an in-crease in controlled processes and conceptualization abilities. These 2 cognitive components might delay the clinical expression of neurodegenerative illnesses by maintaining global cognitive efficiency. Of these 2 components, controlled processes were also influenced by high attention-demanding occupations. PMID- 12740189 TI - Learning disability subtypes and the role of attention during the naming of pictures and words: an event-related potential analysis. AB - The role of attention in the processing of pictures and words was investigated for a group of normally achieving children and for groups of learning disability sub-types that were defined by deficient performance on tests of reading and spelling (Group RS) and of arithmetic (Group A). An event-related potential (ERP) recording paradigm was employed in which the children were required to attend to and name either pictures or words that were presented individually or in superimposed picture-word arrays that varied in degree of semantic relation. For Group RS, the ERP waves to words, both presented individually or attended in the superimposed array, exhibited reduced N450 amplitude relative to controls, whereas their ERP waves to pictures were normal. This suggests that the word naming deficiency for Group RS is not a selective attention deficit but rather a specific linguistic deficit that develops at a later stage of processing. In contrast to Group RS and controls, Group A did not exhibit reliable early frontal negative waves (N280) to the super-imposed pictures and words, an effect that may reflect a selective attention deficit for these children that develops at an early stage of visuo-spatial processing. These early processing differences were also evident in smaller amplitude N450 waves for Group A when naming either pictures or words in the superimposed arrays. PMID- 12740190 TI - Children's attentional skills 2 years post-traumatic brain injury. AB - This study examined sustained attention abilities 24 months postinjury, in 3 groups of children who had suffered a traumatic brain injury (TBI): (a). mild TBI (n = 24), (b). moderate TBI (n = 31), and (c). severe TBI (n = 14). Three manipulations of the traditional Continuous Performance Test paradigm (CPT) were employed in the study, to delineate factors that might influence CPT performance, including speed of processing, attentional lapses, and stimulus complexity. No significant differences were evident among the TBI groups on a measure of simple reaction time, nor on a CPT version where the interstimulus interval was lengthened. However, there was a significant difference between the mild and severe TBI groups on the most complex task, which required speed, accuracy, and decision making, suggesting that these factors underlie impaired performances previously identified on the CPT in children with severe TBI. These findings have implications for the development of intervention programs for these children. PMID- 12740191 TI - Salivary testosterone and EEG spectra of 9- to 11-year-old male children. AB - Although most investigators agree that there are electroencephalogram (EEG) changes with chronological age around puberty, no one has remarked on why this is so. As the increase in testosterone at the end of prepubertal childhood is a biomarker for the on-set of a period of accelerated growth and development, we searched for an association between testosterone level and the spectral power and topography of EEG. We tested 60 children between 9- and 11-years-old, each demonstrating normal neurological examination and Wechsler intelligence scale over 90. Salivary testosterone was measured using immunoenzymatic chemiluminescent assay. Children were divided into 3 groups according to measured testosterone level. EEG was processed with a Fast Fourier Transform; average of relative power spectral analyses were calculated and data divided into delta, theta, alpha, and beta bands. The principal finding was the highly significant interaction between EEG band and testosterone group, F(6, 171) = 4.54, Huynh Feldt epsilon = 0.853, p =.001. Delta relative power decreased significantly and Alpha relative power increased significantly in the 2 groups with higher salivary testosterone concentration when compared to the lowest testosterone group. This work suggests that increased testosterone in prepubertal children can predict some of the same EEG changes that are usually associated with increased chronological age. PMID- 12740192 TI - Speech perception of infants with high familial risk for dyslexia differ at the age of 6 months. AB - As yet relatively little is known of the earliest signs of dyslexia. We present evidence showing that the speech perception of 6-month-old infants from dyslexic families differs significantly from that of infants from control families with normal reading parents; the former group needed a significantly longer duration to categorize speech sounds as long. The same difference appeared in their dyslexic parents. This study shows that differences in categorizing speech sounds according to duration, which is crucial to intelligibility in Finnish, are a factor associated with familial risk for dyslexia already at infancy, which persists until adulthood in many of those suffering from dyslexia. PMID- 12740193 TI - Normal development of bimanual coordination: visuomotor and interhemispheric contributions. AB - The corpus callosum is one of the last cortical pathways to develop, continuing to myelinate through the end of the first decade of life. However, the functional consequences of this late development are not entirely known. The importance of callosal development for bimanual motor coordination is suggested by the fact that bimanual coordination in younger children is similar to that of persons with commissurotomy or callosal agenesis. This study focused on the development of bimanual coordination in 67 normally developing children between 6 and 15 years of age using the computerized Bimanual Coordination Test (cBCT). Results indicated that right- and left-hand unimanual motor speed was significantly correlated with age (r = -.26 and -.44, respectively). Age was also significantly associated with accuracy of performance on trials demanding both symmetric (r = .46) and asymmetric (r = -.50) bi-manual responding. The correlation with asymmetric bimanual responding (requiring greater interhand coordination) remained significant when covarying performance on symmetric response trials. Accuracy on asymmetric bimanual trials requiring greater left- than right-hand speed accounted for the largest portion of this unique, age-related variance. Thus, cBCT performance reveals child development in motor speed and visuomotor processing, as well as the unique contributions of interhemispheric interactions to bimanually coordinated motor activity. PMID- 12740213 TI - Light and water-use efficiencies of pine shoots exposed to elevated carbon dioxide and temperature. AB - An automatic gas exchange system was used to continuously measure water and carbon fluxes of attached shoots of Scots pine trees (Pinus sylvestris L.) grown in environment-controlled chambers for a 3-year period (1998-2000) and exposed to either normal ambient conditions (CON), elevated CO2 (+350 micro mol mol-1; EC), elevated temperature (+2-6 degrees C; ET) or a combination of EC and ET (ECT). EC treatment enhanced the mean daily total carbon flux per unit projected needle area (Fc.d) by 17-21 %, depending on the year. This corresponds to a 16-24 % increase in light-use efficiency (LUE) based on incident photosynthetically active radiation. The EC treatment reduced the mean daily total water flux (Fw.d) by 1-12 %, corresponding to a 13-35 % increase in water-use efficiency (WUE). The ET treatment increased Fc.d by 10-18 %, resulting in an 8-19 % increase in LUE, and Fw.d by 48-74 %, resulting in a reduction of WUE by 19-34 %. There was no interaction between CO2 and temperature elevation in connection with either carbon or water fluxes, as the carbon flux responded similarly in both ECT and EC, while the water flux in the ECT treatment was similar to that in ET. Regressions indicated that the increase in maximum LUE was greater with increasing air temperature, whereas changes in WUE were related only to high vapour pressure deficit. Furthermore, changes in LUE and WUE caused by ECT treatment displayed strong diurnal and seasonal variation. PMID- 12740214 TI - Identification and isolation of mouse type II cells on the basis of intrinsic expression of enhanced green fluorescent protein. AB - The unique morphology and cell-specific expression of surfactant genes have been used to identify and isolate alveolar type II epithelial cells. Because these attributes can change during lung injury, a novel method was developed for detecting and isolating mouse type II cells on the basis of transgenic expression of enhanced green fluorescence protein (EGFP). A line of transgenic mice was created in which EGFP was targeted to type II cells under control of the human surfactant protein (SP)-C promoter. Green fluorescent cells that colocalized by immunostaining with endogenous pro-SP-C were scattered throughout the parenchyma. EGFP was not detected in Clara cell secretory protein-expressing airway epithelial cells or other nonlung tissues. Pro-SP-C immunostaining diminished in lungs exposed to hyperoxia, consistent with decreased expression and secretion of intracellular precursor protein. In contrast, type II cells could still be identified by their intrinsic green fluorescence, because EGFP is not secreted. Type II cells could also be purified from single-cell suspensions of lung homogenates using fluorescence-activated cell sorting. Less than 1% of presorted cells exhibited green fluorescence compared with >95% of the sorted population. As expected for type II cells, ultrastructural analysis revealed that the sorted cells contained numerous lamellar bodies. SP-A, SP-B, and SP-C mRNAs were detected in the sorted population, but T1alpha and CD31 (platelet endothelial cell adhesion molecule) were not, indicating enrichment of type II epithelial cells. This method will be invaluable for detecting and isolating mouse type II cells under a variety of experimental conditions. PMID- 12740212 TI - Aspects of plant intelligence. AB - Intelligence is not a term commonly used when plants are discussed. However, I believe that this is an omission based not on a true assessment of the ability of plants to compute complex aspects of their environment, but solely a reflection of a sessile lifestyle. This article, which is admittedly controversial, attempts to raise many issues that surround this area. To commence use of the term intelligence with regard to plant behaviour will lead to a better understanding of the complexity of plant signal transduction and the discrimination and sensitivity with which plants construct images of their environment, and raises critical questions concerning how plants compute responses at the whole-plant level. Approaches to investigating learning and memory in plants will also be considered. PMID- 12740215 TI - ET-1 activates Ca2+ sparks in PASMC: local Ca2+ signaling between inositol trisphosphate and ryanodine receptors. AB - Ca+ sparks originating from ryanodine receptors (RyRs) are known to cause membrane hyperpolarization and vasorelaxation in systemic arterial myocytes. By contrast, we have found that Ca2+ sparks of pulmonary arterial smooth muscle cells (PASMCs) are associated with membrane depolarization and activated by endothelin-1 (ET-1), a potent vasoconstrictor that mediates/modulates acute and chronic hypoxic pulmonary vasoconstriction. In this study, we characterized the effects of ET-1 on the physical properties of Ca2+ sparks and probed the signal transduction mechanism for spark activation in rat intralobar PASMCs. Application of ET-1 at 0.1-10 nM caused concentration-dependent increases in frequency, duration, and amplitude of Ca2+ sparks. The ET-1-induced increase in spark frequency was inhibited by BQ-123, an ETA-receptor antagonist; by U-73122, a PLC inhibitor; and by xestospongin C and 2-aminoethyl diphenylborate, antagonists of inositol trisphosphate (IP3) receptors (IP3Rs). However, it was unrelated to sarcoplasmic reticulum Ca2+ content, activation of L-type Ca2+ channels, PKC, or cADP ribose. Photorelease of caged-IP3 indicated that Ca2+ release from IP3R could cross-activate RyRs to generate Ca2+ sparks. Immunocytochemistry showed that the distributions of IP3Rs and RyRs were similar in PASMCs. Moreover, inhibition of Ca2+ sparks with ryanodine caused a significant rightward shift in the ET-1 concentration-tension relationship in pulmonary arteries. These results suggest that ET-1 activation of Ca2+ sparks is mediated via the ETA receptor-PLC IP3 pathway and local Ca2+ cross-signaling between IP3Rs and RyRs; in addition, this novel signaling mechanism contributes significantly to the ET-1-induced vasoconstriction in pulmonary arteries. PMID- 12740216 TI - Protein kinase C modifications of VE-cadherin, p120, and beta-catenin contribute to endothelial barrier dysregulation induced by thrombin. AB - The adherens junction is a multiprotein complex consisting of the transmembrane vascular endothelial cadherin (VEC) and cytoplasmic catenins (p120, beta-catenin, plakoglobin, alpha-catenin) responsible for the maintenance of endothelial barrier function. Junctional disassembly and modifications in cadherin/catenin complex lead to increased paracellular permeability of the endothelial barrier. However, the mechanisms of junctional disassembly remain unclear. In this study, we used the proinflammatory mediator thrombin to compromise the barrier function and test the hypothesis that phosphorylation-induced alterations of VEC, beta catenin, and p120 regulate junction disassembly and mediate the increased endothelial permeability response. The study showed that thrombin induced dephosphorylation of VEC, which is coupled to disassembly of cell-cell contacts, but VEC remained in aggregates at the plasma membrane. The cytoplasmic catenins dissociated from the VEC cytoplasmic domain in thin membrane projections formed in interendothelial gaps. We also showed that thrombin induced dephosphorylation of beta-catenin and phosphorylation of p120. Thrombin-induced interendothelial gap formation and increased endothelial permeability were blocked by protein kinase C inhibition using chelerythrine and Go-6976 but not by LY-379196. Chelerythrine also prevented thrombin-induced phosphorylation changes of the cadherin/catenin complex. Thus the present study links posttranslational modifications of VEC, beta-catenin, and p120 to the mechanism of thrombin-induced increase in endothelial permeability. PMID- 12740217 TI - Surfactant protein A is a required mediator of keratinocyte growth factor after experimental marrow transplantation. AB - We reported an association between the ability of recombinant human keratinocyte growth factor (rHuKGF) to upregulate the expression of surfactant protein A (SP A) and to downregulate pulmonary inflammation that occurs after allogeneic bone marrow transplantation (BMT). To establish a causal relationship, rHuKGF (5 mg/kg) was administered subcutaneously for three consecutive days before irradiation to SP-A-sufficient and -deficient [SP-A(+/+) and SP-A(-/-), respectively] mice given inflammation-inducing allogeneic spleen T cells at the time of BMT. In contrast with SP-A(+/+) mice, rHuKGF failed to suppress the high levels of TNF-alpha, IFN-gamma, and nitric oxide contained in bronchoalveolar lavage fluids collected on day 7 after BMT from SP-A(-/-) mice. Early post-BMT weight loss was attenuated by rHuKGF in both SP-A(+/+) and SP-A(-/-) recipients. In the absence of supportive respiratory care, however, SP-A deficiency eventually abolished the ability of rHuKGF to prevent weight loss and to improve survival monitored for 1 mo after allogeneic BMT. In further experiments, the addition of cyclophosphamide (which is known to cause severe injury to the alveolar epithelium in donor T cell-recipient mice) to the conditioning regimen prevented rHuKGF-induced upregulation of SP-A and suppression of lung inflammation in both SP-A(+/+) and SP-A(-/-) mice. We conclude that endogenous baseline SP-A levels and optimal upregulation of SP-A are required for the anti inflammatory protective effects of KGF after allogeneic transplantation. PMID- 12740218 TI - Bone morphogenetic proteins induce apoptosis in human pulmonary vascular smooth muscle cells. AB - Pulmonary vascular medial hypertrophy in primary pulmonary hypertension (PPH) is mainly caused by increased proliferation and decreased apoptosis in pulmonary artery smooth muscle cells (PASMCs). Mutations of the bone morphogenetic protein (BMP) receptor type II (BMP-RII) gene have been implicated in patients with familial and sporadic PPH. The objective of this study was to elucidate the apoptotic effects of BMPs on normal human PASMCs and to examine whether BMP induced effects are altered in PASMCs from PPH patients. Using RT-PCR, we detected six isoforms of BMPs (BMP-1 through -6) and three subunits of BMP receptors (BMP-RIa, -RIb, and -RII) in PASMCs. Treatment of normal PASMCs with BMP-2 or -7 (100-200 nM, 24-48 h) markedly increased the percentage of cells undergoing apoptosis. The BMP-2-mediated apoptosis in normal PASMCs was associated with a transient activation or phosphorylation of Smad1 and a marked downregulation of the antiapoptotic protein Bcl-2. In PASMCs from PPH patients, the BMP-2- or BMP-7-induced apoptosis was significantly inhibited compared with PASMCs from patients with secondary pulmonary hypertension. These results suggest that the antiproliferative effect of BMPs is partially due to induction of PASMC apoptosis, which serves as a critical mechanism to maintain normal cell number in the pulmonary vasculature. Inhibition of BMP-induced PASMC apoptosis in PPH patients may play an important role in the development of pulmonary vascular medial hypertrophy in these patients. PMID- 12740219 TI - Phorbol esters increase MLC phosphorylation and actin remodeling in bovine lung endothelium without increased contraction. AB - Direct protein kinase C (PKC) activation with phorbol myristate acetate (PMA) results in the loss of endothelial monolayer integrity in bovine lung endothelial cells (EC) but produces barrier enhancement in human lung endothelium. To extend these findings, we studied EC contractile events and observed a 40% increase in myosin light chain (MLC) phosphorylation in bovine endothelium following PMA challenge. The increase in PMA-mediated MLC phosphorylation occurred at sites distinct from Ser19/Thr18, sites catalyzed by MLC kinase (MLCK), and immunoblotting with antibodies specific to phosphorylated Ser19/Thr18 demonstrated profound time-dependent Ser19/Thr18 dephosphorylation. These events occurred in conjunction with rearrangement of stress fibers into a grid-like network, but without an increase in cellular contraction as measured by silicone membrane wrinkling assay. The PMA-induced MLC dephosphorylation was not due to kinase inhibition but, rather, correlated with rapid increases in myosin associated phosphatase 1 (PPase 1) activity. These data suggest that PMA-mediated EC barrier regulation may involve dual mechanisms that alter MLC phosphorylation. The increase in bovine MLC phosphorylation likely occurs via direct PKC-dependent MLC phosphorylation in conjunction with decreases in Ser19/Thr18 phosphorylation catalyzed by MLCK due to PMA-induced increases in PPase 1 activity. Together, these events result in stress fiber destabilization and profound actin rearrangement in bovine endothelium, which may result in the physiological alterations observed in these models. PMID- 12740220 TI - Thyroid hormone stimulates Na-K-ATPase activity and its plasma membrane insertion in rat alveolar epithelial cells. AB - Na-K-ATPase protein is critical for maintaining cellular ion gradients and volume and for transepithelial ion transport in kidney and lung. Thyroid hormone, 3,3',5 triiodo-l-thyronine (T3), given for 2 days to adult rats, increases alveolar fluid resorption by 65%, but the mechanism is undefined. We tested the hypothesis that T3 stimulates Na-K-ATPase in adult rat alveolar epithelial cells (AEC), including primary rat alveolar type II (ATII) cells, and determined mechanisms of the T3 effect on the Na-KATPase enzyme using two adult rat AEC cell lines (MP48 and RLE-6TN). T3 at 10-8 and 10-5 M increased significantly hydrolytic activity of Na-K-ATPase in primary ATII cells and both AEC cell lines. The increased activity was dose dependent in the cell lines (10-9-10-4 M) and was detected within 30 min and peaked at 6 h. Maximal increases in Na-K-ATPase activity were twofold in MP48 and RLE-6TN cells at pharmacological T3 of 10-5 and 10-4 M, respectively, but increases were statistically significant at physiological T3 as low as 10-9 M. This effect was T3 specific, because reverse T3 (3,3',5'-triiodo-l thyronine) at 10-9-10-4 M had no effect. The T3-induced increase in Na-K-ATPase hydrolytic activity was not blocked by actinomycin D. No significant change in mRNA and total cell protein levels of Na-K-ATPase were detected with 10-9-10-5 M T3 at 6 h. However, T3 increased cell surface expression of Na-K-ATPase alpha1- or beta1-subunit proteins by 1.7- and 2-fold, respectively, and increases in Na-K ATPase activity and cell surface expression were abolished by brefeldin A. These data indicate that T3 specifically stimulates Na-K-ATPase activity in adult rat AEC. The upregulation involves translocation of Na-K-ATPase to plasma membrane, not increased gene transcription. These results suggest a novel nontranscriptional mechanism for regulation of Na-K-ATPase by thyroid hormone. PMID- 12740221 TI - Inflammation and vascular hypertrophy induced by angiotensin II: role of NADPH oxidase-derived reactive oxygen species independently of blood pressure elevation? PMID- 12740222 TI - Role of ABCA1 in cellular cholesterol efflux and reverse cholesterol transport. PMID- 12740224 TI - Combinatorial control of smooth muscle-specific gene expression. AB - Alterations in the differentiated state of vascular smooth muscle cells (SMCs) are known to play a key role in vascular diseases, yet the mechanisms controlling SMC differentiation are still poorly understand. In this review, we discuss our present knowledge of control of SMC differentiation at the transcriptional level, pointing out some common themes, important paradigms, and unresolved issues in SMC-specific gene regulation. We focus primarily on the serum response factor CArG box-dependent pathway, because it has been shown to play a critical role in regulation of multiple SMC marker genes. However, we also highlight several other important regulatory elements, such as a transforming growth factor beta control element, E-boxes, and MCAT motifs. We present evidence in support of the notion that SMC-specific gene regulation is not controlled by a few SMC-specific transcription factors but rather by complex combinatorial interactions between multiple general and tissue-specific proteins. Finally, we discuss the implications of chromatin remodeling on SMC differentiation. PMID- 12740225 TI - Rapid analysis of angiogenesis drugs in a live fluorescent zebrafish assay. PMID- 12740226 TI - Confirmation of the microsomal triglyceride transfer protein genetic effect on lipids in young African American men from the CARDIA study. PMID- 12740227 TI - Possible impact of tetrahydrobiopterin and sepiapterin on endothelial dysfunction. PMID- 12740228 TI - The problems and challenges of immunotherapy in sepsis. AB - Despite decades of research, the morbidity and mortality of sepsis and septic shock remain very high. To further compound the problem, results from all investigative trials (with one exception) have shown that tested immunotherapies aimed at modulating the excessive expression of key cytokines, such as the interleukins and tumor necrosis factor, have been either equivalent or inferior to placebo. While controversy prevails in terms of continuing such investigative trials, study designs can be held accountable for inherent flaws. Testing for the wrong hypothesis, errant study design, using the wrong agent, focusing on an inappropriate target group, excessive expectations, and uncontrolled variables have potentially obscured the real efficacy such agents might have to offer. By standardizing protocols and reducing uncontrolled variables, research can be more precisely targeted so as to unmask the real benefits to the patient. PMID- 12740229 TI - Dopexamine, dobutamine, and dopamine increase splanchnic blood flow: what is the evidence? AB - Inadequate splanchnic perfusion in the critically ill compromises the gut barrier leading to bacterial translocation, which is postulated to cause multiorgan dysfunction and failure. Inotropic agents such as dopexamine, dobutamine, and dopamine may have a role in increasing splanchnic perfusion, thereby protecting this area from further injury. This article examines the evidence for using these agents in patients with sepsis, postoperative trauma, and in those undergoing cardiac surgery and mechanical ventilation to increase gut perfusion and prevent multiple organ failure. Systemic effects of these agents differ from regional effects and must be considered when selecting therapy. PMID- 12740230 TI - Selective digestive decontamination should not be routinely employed. AB - There is a general consensus that antimicrobial resistance in the hospital setting has emerged as an important variable influencing patient outcome and resource utilization. Hospitals worldwide are faced with increasingly rapid emergence and spread of antibiotic-resistant bacteria. Both antibiotic-resistant Gram-negative bacilli and Gram-positive bacteria are reported as important causes of hospital-acquired infections. Few antimicrobial agents are available for effective treatment. Selective digestive decontamination (SDD) is a technique aimed at selectively eliminating aerobic Gram-negative bacilli and yeast from the mouth and stomach to reduce the occurrence of hospital-acquired infections, including ventilator-associated pneumonia. Unfortunately, the application of SDD has been associated with emergence of antibiotic-resistant bacterial strains, limiting its overall utility. PMID- 12740231 TI - Gastric tonometry: the hemodynamic monitor of choice (Pro). AB - Controversy exists as to the best means to monitor the critically ill patient and the appropriate end points of therapy. Use of global hemodynamic or metabolic parameters may be normal in the patient who has not been completely or adequately resuscitated. Decreased perfusion to the gut is not well tolerated and may contribute to the development of the multiple organ dysfunction syndrome. Gastric tonometry is a minimally invasive way to monitor splanchnic perfusion in the critically ill patient. Data suggest that tonometry is useful for outcome prognostication and for detection of early hypovolemia. In addition, use of gastric intramucosal pH or mucosal-arterial CO(2) gap as end points of resuscitation may be superior to other conventional whole-body parameters. For these reasons, gastric tonometry must be considered the hemodynamic monitor of choice. PMID- 12740232 TI - Gastric tonometry: the role of mucosal pH measurement in the management of trauma. AB - Effective management of hemorrhagic shock depends on titration of therapies against reliable resuscitation end points. Conventional clinical and laboratory indexes of shock are often slow to respond to progressive circulatory compromise. GI mucosal ischemia resulting from redistribution of blood flow may, however, precede uncompensated shock and may compound the initial hemorrhagic insult by touching off cascades of inflammatory responses. Trauma patients with evidence of subclinical GI ischemia have been shown to have poor outcomes. Gastric tonometry, by detecting the presence of gastric intramucosal acidosis as a proxy of splanchnic hypoperfusion, may facilitate more timely and rational shock resuscitation. This article reviews the development and validation of gastric tonometry and summarizes the clinical studies that have used this modality to guide the management of shock in trauma patients. PMID- 12740233 TI - Steroids for septic shock: back from the dead? (Con). AB - The role of corticosteroid therapy in the management of septic shock has been debated for half a century. Results from large, well-designed, randomized clinical trials demonstrate no benefit, and perhaps harm, associated with short duration, high-dose methylprednisolone or dexamethasone administered at the onset of septic shock. Based on evidence of "relative adrenal insufficiency" and steroid-responsive adrenergic receptor desensitization in sepsis, administration of modest doses (200 to 300 mg/d) of hydrocortisone for 1 to 3 weeks has been investigated. A multicenter, placebo-controlled clinical trial demonstrated improved survival rates and faster cessation of vasopressors among patients with septic shock who have a poor response to corticotropin injection, consistent with relative adrenal insufficiency. However, concerns regarding a trend for higher mortality among corticotropin responders and the possibility that patients with true adrenal insufficiency may have been enrolled in this placebo-controlled trial, potentially skewing results, should be considered. PMID- 12740234 TI - Steroids for septic shock: back from the dead? (Pro). AB - The use of corticosteroids as adjunctive therapy for severe sepsis and septic shock has been a source of controversy for the past 35 years. Despite a wealth of preclinical data supporting both survival and physiologic benefit for early steroid use, the data in human sepsis have been much less convincing. There have even been reports suggesting the potential for harm associated with the administration of early high-dose corticosteroids in patients with severe sepsis and septic shock. Recent trials have reported hemodynamic and survival benefits associated with the use of more physiologic steroid replacement therapy in patients with vasopressor-dependent septic shock. These results coupled with the observation of "relative adrenal insufficiency" in some patients with severe sepsis and septic shock may once again establish a defined role for corticosteroid therapy in the management of severe sepsis and septic shock. PMID- 12740235 TI - Shifting patterns in the epidemiology of nosocomial Candida infections. AB - The incidence of candidemia--a common and potentially fatal nosocomial infection- has risen dramatically, and this increase has been accompanied by a shift in the infecting pathogen away from Candida albicans to treatment-resistant non-albicans species. Prophylactic azole antifungals, such as fluconazole, may play an important role not only in the management of candidemia but also in the proliferation of hard-to-treat Candida species. In a variety of acute nosocomial settings, IV fluconazole, 400 mg/d, has reduced Candida colonization and infection. A growing body of evidence supports the still controversial contention that the increasing use of azole antifungals is at least partially responsible for the proliferation of treatment-resistant, non-albicans isolates, especially Candida glabrata. Thus, selecting the most appropriate candidates for prophylactic antifungal intervention--ie, those with the highest risk for candidemia--may be indispensable, not only in preventing candidemia, but also in reducing antifungal overuse, which may contribute to the emergence of treatment resistant Candida isolates. PMID- 12740236 TI - Vancomycin-resistant enterococci: a road map on how to prevent the emergence and transmission of antimicrobial resistance. AB - Nosocomial acquisition of microorganisms resistant to multiple antibiotics represents a threat to patient safety. Here we review the mechanisms that have allowed highly resistant strains belonging to the Enterococcus genus to proliferate within our health-care institutions. These mechanisms indicate that decreasing the prevalence of resistant organisms requires active surveillance, adherence to vigorous isolation, hand hygiene and environmental decontamination measures, and effective antibiotic stewardship. We suggest how to tailor such a complex, multidisciplinary program to the needs of a particular health-care setting so as to maximize cost-effectiveness. PMID- 12740237 TI - Non-small cell lung cancer with chest wall involvement. PMID- 12740238 TI - Outcome measurements in COPD: are we schizophrenic? PMID- 12740239 TI - Combination antibiotic therapy with macrolides in community-acquired pneumonia: more smoke but is there any fire? PMID- 12740240 TI - The variability of sarcoidosis: can we predict it? PMID- 12740241 TI - The conundrum of sleep breathing disorders in heart failure. PMID- 12740242 TI - Prone position in ARDS: are we looking at a half-empty or half-full glass? PMID- 12740243 TI - Percutaneous dilatational tracheostomy: we live in a twisted world. PMID- 12740244 TI - Smoking: not for anyone. PMID- 12740245 TI - Non-small cell lung cancer with chest wall invasion: evolution of surgical treatment and prognosis in the last 3 decades. AB - STUDY OBJECTIVES: The treatment of patients with non-small cell lung cancer (NSCLC) that is invading the chest wall is still debated. We aim to illustrate the improvements in treatment results that have occurred over last decade. DESIGN: Retrospective analysis of our experience and an overview of the literature. SETTING: Department of Surgery, San Giuseppe Hospital, University of Milan. PATIENTS: From January 1970 to December 1999, of 2,738 patients with NSCLC, we operated on 146 patients (5.4%) with chest wall invasion by NSCLC. Superior sulcus tumors and tumors invading the diaphragm or mediastinum were excluded. We reclassified all cases according to the current TNM classification. RESULTS: We registered one postoperative death (0.69%) and five major complications (3.4%). From 1970 to 1979, of 32 patients, 10 underwent an exploratory thoracotomy (ET) and 22 underwent a radical resection (stage IIB disease, 17 patients; stage IIIA disease, 5 patients). The 5-year survival rate was 22.7% (25% for stage IIB disease). From 1980 to 1989, of 67 patients, 11 underwent an ET and 56 underwent a radical resection (stage IIB disease, 34 patients; stage IIIA disease, 12 patients; stage IIIB disease, 5 patients; and stage IV disease, 5 patients). The survival rate following radical resection was 14.1%, ranging between 23.5% for patients with stage IIB disease and 0% (3 years, 14%) for those with stage IIIA disease. From 1990 to 1999, of 47 patients, 2 underwent an ET, 2 underwent an exploratory thoracoscopy, and 43 underwent a radical resection (stage IIB disease, 23 patients; stage IIIA disease, 20 patients). The survival rate was 42.7% (stage IIB disease, 78.5%; stage IIIA disease, 7.2%). CONCLUSIONS: Considering the low morbidity, mortality, and significant improvement in survival during the last decade, we advocate the performance of radical en bloc resection for the treatment of chest wall invasive NSCLC. PMID- 12740246 TI - Technical aspects of composite arterial grafting with double skeletonized internal thoracic arteries. AB - BACKGROUND: Complete myocardial revascularization with internal thoracic arteries (ITAs) improves long-term survival and decreases the rate of repeat operations, compared to vein grafts. Adequate length of the graft in coronary artery bypass graft (CABG) surgery is essential for providing complete arterial revascularization. Extra length can be obtained by skeletonization of both ITAs. In cases where the right ITA (RITA) is too short to bridge the distance to the target anastomotic site, it is used as a free graft in "composite" arterial grafting, a surgical technique in which free arterial conduits are proximally anastomosed end-to-side to an intact ITA. OBJECTIVES: To describe alternative surgical procedures adapted to accommodate special anatomic requirements. DESIGN: Retrospective study from April 1996 to April 1999. PATIENTS: One thousand fifty patients underwent CABG surgery using bilateral skeletonized ITAs: 650 patients (482 men and 168 women; mean +/- SD age, 69 +/- 7 years) underwent composite arterial grafting. Two hundred sixteen patients (33.2%) were diabetics, 87 patients (13.4%) had severe left ventricular dysfunction (ejection fraction < 35%), and 27 patients (4.2%) underwent emergency operations. INTERVENTIONS: The RITA was used as a free graft connected to the in situ left ITA (LITA) in 618 patients. A free LITA was attached to in situ RITA in 32 patients, and minicomposite grafts (free distal LITA on the LITA or free distal RITA on the RITA) were constructed in 38 patients. The average number of grafts was 3.2 per patient (range, 2 to 6 grafts per patient). MEASUREMENTS AND RESULTS: Operative mortality was 2.9% (n = 19), and there were 11 sternal wound infections (1.7%). Early recatheterization was performed in 41 symptomatic patients. The patency rate was 95%. The mean follow-up was 25 months (range, 14 to 36 months), and the 3-year survival was 92.5%, with 97% of the surviving patients being angina free. CONCLUSIONS: Planning CABG surgery using bilateral skeletonized ITAs as arterial conduits affords greater choice in grafting approaches, especially when a composite technique is feasible. PMID- 12740247 TI - Transcardiac release of soluble adhesion molecules during coronary artery bypass grafting: effects of crystalloid and blood cardioplegia. AB - BACKGROUND: Dysfunction of myocardium as a result of ischemia/reperfusion during coronary artery bypass grafting (CABG) is currently one of the biggest problems in cardiovascular surgery. In previous studies, it has been well established that activated leukocytes and coronary vascular endothelial cells play an important role in the development of cardiac tissue damage during ischemia followed by reperfusion. Interactions between both of these cell types require the expression of adhesion molecules on their surface. In certain conditions, on cell activation, the adhesion proteins may be released from activated cells in soluble form into circulation. The purpose of our study was to establish whether the use of blood cardioplegia modifies plasma levels of soluble intracellular adhesion molecule-1 (sICAM-1), soluble vascular cell adhesion molecule-1 (sVCAM-1), soluble E-selectin (sE-selectin), and soluble L-selectin (sL-selectin) in comparison with crystalloid cardioplegia in patients undergoing CABG. METHODS: Patients undergoing CABG were classified into two groups to receive cold crystalloid cardioplegia (St. Thomas' Hospital) or cold blood cardioplegia (method of Buckberg), followed by a "warm-shot" of the solution. Coronary sinus and arterial blood samples were obtained from 50 patients (42 men and 8 women; age range, 34 to 73 years) before aortic cross-clamping, at the beginning of reperfusion, and after 30 min of reperfusion. Plasma levels of soluble adhesion molecules were measured using sensitive enzyme-linked immunosorbent assays. RESULTS: The transcardiac release of sICAM-1 and sVCAM-1 following myocardial ischemia/reperfusion during CABG was evident in both groups of patients. However, the increase of soluble forms of both of these adhesion proteins was more significant in the group of patients receiving crystalloid cardioplegia. Crystalloid cardioplegia resulted in decreased plasma level of sE-selectin in the coronary sinus blood sample after 30 min of reperfusion. No significant changes in plasma levels of sL-selectin in either group were observed. CONCLUSION: Cardioplegia may affect the release of soluble forms of adhesion molecules from ischemic myocardium and modify endothelium activation in patients undergoing CABG. PMID- 12740248 TI - Frequency, risk factors, and outcome of hyperlactatemia after cardiac surgery. AB - STUDY OBJECTIVE: To determine the respective frequencies, risk factors, and outcomes of no hyperlactatemia (NHL), immediate hyperlactatemia (IHL), or late hyperlactatemia (LHL) > 3 mmol/L after cardiac surgery. DESIGN: Prospective and observational study. SETTING: Cardiac surgery ICU in a 130-bed private community nonteaching hospital. PATIENTS: Consecutive patients (n = 325) undergoing cardiopulmonary bypass (CPB) for cardiac surgery. INTERVENTION: None. MEASUREMENTS: Arterial blood gas levels and lactate concentrations were measured at ICU admission, 4 h after surgery, between 6 h and 16 h after surgery, and on day 1. MAIN RESULTS: Sixty-seven patients (20.6%) had an IHL on ICU admission, and 56 patients (17.2%) acquired LHL during their ICU stay. ICU mortality was 1.5% for NHL, 3.6% for LHL, and 14.9% for IHL groups (p < 0.0001). The three groups differed significantly for elective surgery, type of operation, CPB duration, intraoperative mean arterial pressure, and intraoperative and postoperative use of vasopressor. Independent risk factors for IHL were nonelective surgery, CPB duration, and intraoperative use of vasopressor. Logistic regression identified hyperglycemia and epinephrine therapy for LHL as postoperative risk factors. Receiver operating characteristic curves showed that IHL more accurately predicted ICU mortality than LHL. CONCLUSIONS: Hyperlactatemia is common after cardiac surgery. A lactate threshold of 3 mmol/L at ICU admission is able to identify a population at risk of morbidity and mortality after cardiac surgery. PMID- 12740249 TI - Recovery of self-reported functional capacity after coronary artery bypass surgery. AB - STUDY OBJECTIVES: This study was conducted to determine the effects of age, gender, comorbid conditions, and exercise on the recovery of self-reported functional capacity after coronary artery surgery, and to identify predictors of 1-year functional capacity. PATIENTS: One hundred ninety-eight patients undergoing coronary artery bypass graft surgery. MEASUREMENTS AND RESULTS: Self reported functional capacity was evaluated before surgery, and 3 months and 1 year postoperatively using the Veterans Specific Activity Questionnaire (VSAQ). Patients were classified into groups based on age, gender, comorbid conditions, and postoperative exercise. Repeated-measures analysis of variance was used to determine if groups differed with respect to functional capacity recovery and multiple linear regression was used to identify predictors of 1-year VSAQ score. A significant time by age interaction was found (p = 0.0001), with a more protracted recovery for older patients. There were significant group effects for gender (p = 0.0001), and presence of comorbid conditions (p = 0.0009); however, there were no time/group interactions for these variables. A significant group effect was found for postoperative exercise (p = 0.0001), with a trend toward group/time interaction (p = 0.096). Predictors of 1-year functional capacity were VSAQ score in the year prior to surgery and performance of regular aerobic exercise in the postoperative period. CONCLUSIONS: This study suggests that older patients attain good self-reported functional outcomes after surgery; however, the time course for recovery is more protracted than for younger patients. Functional capacity in the year prior to surgery and postoperative exercise are key predictors of 1-year functional capacity. PMID- 12740250 TI - D-dimer in acute aortic dissection. AB - STUDY OBJECTIVE: Laboratory testing plays a minor role in the assessment of aortic dissection. Its main value is in the exclusion of other diseases. Following an incidental observation, we systematically investigated the relationship between elevated d-dimer levels and acute aortic dissection. DESIGN: We prospectively tested d-dimer levels in patients with suspected acute aortic dissection (10 patients). In addition, we investigated 14 patients who had received a confirmed diagnosis of thoracic aortic dissection during the previous 5 years, in whom d-dimer testing had been performed for differential diagnosis. Thirty-five patients with acute chest pain of other origin served as a control group. SETTING: Tertiary referral hospital. PATIENTS: Twelve patients had type A dissection (Stanford classification), and 12 patients had type B. MEASUREMENTS AND RESULTS: A d-dimer analysis was performed (Tina-quant assay; Roche Diagnostics; Mannheim, Germany) [normal limit of the assay, 0.5 micro g/mL]. The result of the d-dimer test was positive (ie, > 0.5 micro g/mL) in all patients (sensitivity of the test, 100%) with a mean value of 9.4 micro g/mL and a range of 0.63 to 54.7 micro g/mL. The degree of the elevation was correlated to the delay from the onset of symptoms to laboratory testing (mean, 12.6 h; range, 1 to 120 h) and showed a trend to the extent of the dissection, but not to the outcome (14 patients could be discharged; 10 patients died). CONCLUSIONS: Based on our observation, we suggest that testing for d-dimer should be part of the initial assessment of patients with chest pain, especially if aortic dissection is suspected. A negative test result makes the presence of the disease unlikely. PMID- 12740251 TI - Clinicopathologic study and DNA analysis of 37 cardiac myxomas: a 28-year experience. AB - PURPOSE: This study was performed to identify morphologic features of cardiac myxomas related to embolism and to provide a better understanding of the biology of these tumors, mainly in relation to their interleukin (IL)-6 expression and DNA content. PATIENTS AND METHODS: A total of 37 cardiac myxomas were reviewed retrospectively in a clinicopathologic study that included the correlation of echocardiographic and pathologic findings in 25 cases, together with immunohistochemical evaluation of IL-6 expression and flow cytometric DNA analysis of 35 tumors. RESULTS: There were 24 female patients and 13 male patients. The mean (+/- SD) age was 52 +/- 15 years. Fifty-four percent of patients presented with dyspnea, 51% presented with increased erythrocyte sedimentation rate (ESR), and 27% presented with embolic episodes, which were significantly associated with villous surface tumors. Atrial fibrillation was registered in 19% of patients and was significantly associated with large left atrial myxomas. Echocardiography proved to be a reliable method for preoperative diagnosis and for predicting tumor size and morphology. There was no perioperative mortality or long-term recurrences. The frequency of early surgical complications was associated with a longer mean ischemic time. Seventeen percent of tumors had abnormal DNA content, and 74% of tumors showed immunohistochemical expression of IL-6. Neither of these factors showed a significant association with embolism or constitutional illness. CONCLUSIONS: Villous surface myxomas are related to embolism, and large left atrial tumors are related to atrial fibrillation. Echocardiography is a reliable method with which to predict tumor size and morphology. Myxoma cells usually express IL-6, and some tumors have abnormal cellular DNA content. Surgical excision of the tumor is a safe and effective treatment. PMID- 12740252 TI - Resting lung function and hemodynamic parameters as predictors of exercise capacity in patients with chronic heart failure. AB - STUDY OBJECTIVES: The aim of this study was to examine the role of resting pulmonary function and hemodynamic parameters as predictors of exercise capacity in patients with chronic heart failure. MEASUREMENTS AND RESULTS: Fifty-one patients with chronic heart failure underwent resting pulmonary function testing, including inspiratory capacity (IC) and symptom-limited, treadmill cardiopulmonary exercise testing (CPET). Right-heart catheterization and radionuclide ventriculography were performed within 2 days of CPET. Mean (+/- SD) left ventricular ejection fraction was 31 +/- 12% and cardiac index was 2.34 +/- 0.77 L/min/m(2). Percentage of predicted FEV(1) was 92 +/- 14%, percentage of predicted FVC was 94 +/- 15%, FEV(1)/FVC was 81 +/- 4%, and percentage of predicted IC was 84 +/- 18%. Mean peak oxygen uptake (peak O(2)) was 17.9 +/- 5.4 mL/kg/min. Analysis of variance among the three functional Weber classes showed statistically significant differences for pulmonary capillary wedge pressure (PCWP) and IC. Specifically, the more severe the exercise intolerance, the lower was IC and the higher was PCWP. In a multivariate stepwise regression analysis, using peak O(2) (liters per minute) as the dependent variable and the pulmonary function test measurements as independent variables, the only significant predictor selected was IC (r = 0.71, p < 0.0001). In a final stepwise regression analysis including all the independent variables of the resting pulmonary function tests and hemodynamic measurements, the two predictors selected were IC and PCWP (r(2) = 0.58). CONCLUSIONS: In patients with chronic heart failure, IC is inversely related to PCWP and is a strong independent predictor of functional capacity. PMID- 12740253 TI - Single-breath diffusing capacity of the lung for carbon monoxide: a predictor of PaO2, maximum work rate, and walking distance in patients with emphysema. AB - OBJECTIVE: The National Emphysema Treatment Trial (NETT) is a randomized, multicenter, clinical trial comparing two different methods of lung volume reduction surgery plus medical therapy to medical treatment alone in patients with advanced emphysema. The purpose of this article was to use the data obtained from the NETT to assess the ability of the single-breath diffusing capacity of the lung for carbon monoxide (DLCO) to predict the need for supplemental oxygen during rest and exercise, as well as overall exercise capacity. METHODS: One thousand seventy-one patients with a mean (+/- SD) FEV(1) of 0.76 +/- 0.24 L were studied. RESULTS: The mean DLCO was 8.0 +/- 3.1 mL/min/mm Hg (28 +/- 10% of predicted). The mean resting PaO(2) was 64 +/- 10 mm Hg. There was a positive association between DLCO and both resting PaO(2) and the requirement for oxygen during a walk at 1 mile per hour (mph). The odds of requiring supplemental oxygen while walking at 1 mph was nine times greater in patients with a DLCO of < or = 20% of predicted than for patients with a DLCO of > 35% of predicted, after adjusting for age and gender. Eighty four percent of individuals with a DLCO of < or = 20% required supplemental oxygen with low levels of exercise compared to 38% of those with a DLCO of > 35%. CONCLUSION: Our results demonstrated that patients with reduced DLCO, particularly when < or = 20% of predicted, are more likely to have reduced PaO(2) at rest and are more likely to require supplemental oxygen with low levels of activity. Thus, DLCO is useful in evaluating whether supplemental oxygen is required for exercise. PMID- 12740254 TI - 6-minute walk testing is more sensitive than maximal incremental cycle testing for detecting oxygen desaturation in patients with COPD. AB - STUDY OBJECTIVES: Some respiratory patients exhibit oxygen desaturation during rehabilitative walking but not during maximal cardiopulmonary exercise testing (CPET). We evaluated exercise-induced desaturation during 6-min walk testing (6MWT) in comparison with CPET in patients with COPD and determined the reproducibility of the phenomenon. PATIENTS: We tested 80 consecutive patients with COPD (FEV(1), 62.4 +/- 2% predicted) and 10 patients with supplementary COPD (FEV(1), 59.1 +/- 5% predicted) [mean +/- SEM] to determine the reproducibility. MEASUREMENTS AND RESULTS: First, patients with COPD performed cycle CPET (first CPET [CPET-1]). Three days later, they performed two 6MWTs (first 6MWT [6MWT-1] and second 6MWT [6MWT-2]). Pulse oximetric saturation (SpO(2)) was recorded every minute in both tests. Three groups emerged: desaturation at 6MWT not observed at CPET (DND) [n = 23], desaturation in both tests (n = 16), and no desaturation in either test (n = 41). Second, to evaluate reproducibility, 10 additional subjects with COPD who exhibited desaturation during two successive 6MWTs but not in CPET performed a second CPET (CPET-2) and a single-bout 6MWT (6MWT-3) in a supplementary trial. When two CPETs were performed, lack of O(2) desaturation was noted in both. O(2) desaturation was confirmed in 6MWT-2 and 6MWT-3 (7.4 +/- 1% and 7.4 +/- 1.5%, respectively). CONCLUSION: Twenty-eight percent of patients with COPD presented DND. The phenomenon was reproducible and not protocol dependent, emphasizing the clinical interest of the 6MWT. PMID- 12740255 TI - 6-minute walk work for assessment of functional capacity in patients with COPD. AB - The 6-min walk (6MW) test is commonly used to assess exercise capacity in patients with COPD and to track functional change resulting from disease progression or therapeutic intervention. Not surprisingly, distance covered has been the preferred outcome for this test. However, distance walked does not account for differences in body weight that are known to influence exercise capacity. OBJECTIVE: The aim of this study was to evaluate the 6-min distance x body weight product (6MWORK) as an improved outcome measure with a solid physiologic foundation. PATIENTS AND METHODS: One hundred twenty-four men and women with moderate-to-severe COPD volunteered and completed the testing sequence, which included pulmonary function, a peak effort ramp cardiopulmonary exercise study with gas exchange, and the 6MW. Means and SD were generated for the variables of interest. Differences were analyzed using analysis of variance techniques. Correlation coefficients and receiver operating characteristic (ROC) curves were calculated for the 6-min walk distance (6MWD) and 6MWORK with indexes of pulmonary function, work performance, and Borg scores for dyspnea and effort. RESULTS: Men and women presented with a significant smoking history that also differed by gender (48 vs 66 pack-years, respectively; p < 0.01). The mean (+/- SD) FEV(1) values were 45 +/- 12.6% and 48 +/- 12.1%, respectively (not significant), while the diffusing capacity of the lung for carbon monoxide (DLCO) was 14.7 +/- 6.1 vs 10.3 +/- 3.9 mL/min/mm Hg, respectively (p < 0.001), for men and women. The 6MWD averaged 416.8 +/- 79.0 m for men and 367.8 +/- 78.6 m for women, and these differences were significant (p < 0.002). When 6MWD was compared as the percent predicted of normal values, each gender presented with a similar reduction of 78.6 +/- 14.5% vs 79.9 +/- 17.5% (p > 0.05), respectively. 6MWORK averaged 35,370 +/- 9,482 kg/m and 25,643 +/- 9,080 kg/m (p < 0.0001) for men and women, respectively. 6MWORK yielded higher correlation coefficients than did 6MWD when correlated with DLCO, lung diffusion for alveolar ventilation, FEV(1), FEV(1)/FVC ratio, watts, peak oxygen uptake, peak minute ventilation, and peak tidal volume. The ROC curve demonstrated that 6MWORK had a significantly larger calculated area under the curve (p < 0.05) [plot of 100-sensitivity to specificity for each variable of interest for all subjects] than 6MWD when differentiating an objectively selected definition of low work capacity vs high work capacity (bike ergometry work, < 55 vs > 55 W, respectively). CONCLUSIONS: We conclude that work calculated as the product of distance x body weight is an improved outcome measure for the 6MW. 6MWORK can be used whenever the 6MW is required to estimate a patient's functional capacity. This measure is also a common measure, which can be converted to indexes of caloric expenditure for direct cross-modality comparisons. PMID- 12740256 TI - Striking similarities in systemic factors contributing to decreased exercise capacity in patients with severe chronic heart failure or COPD. AB - AIMS: Chronic heart failure (CHF) and COPD are both characterized by muscular impairment. To assess whether the severity and functional consequences of muscular impairment are disease specific, we compared skeletal muscle function, body composition, and daily activity level relative to exercise capacity between these two disorders. METHODS: Twenty-five patients with CHF and 25 patients with COPD, and 36 healthy gender- and age-matched control subjects underwent measurement of fat-free mass (FFM) [by bioelectrical impedance analysis] as an index of muscle mass. Quadriceps and biceps functions were tested by isokinetic methods, and daily activity level was assessed by the Physical Activity Scale for Elderly (PASE) questionnaire. Peak oxygen consumption (O(2)peak) was measured by incremental cycle ergometry. RESULTS: PASE results were similar in patients with CHF and in patients with COPD, each group scoring lower than control subjects. FFM was also lower in patients than control subjects and correlated closely with quadriceps and biceps strength in all three subgroups, R values ranging from 0.63 to 0.78, with identical slopes. FFM also correlated significantly with O(2)peak (p < 0.05), but slopes were less steep in patients than in control subjects. The type and severity of muscle dysfunction were similar in each group of patients. There were no significant correlations between indexes of cardiopulmonary function and muscle function or exercise performance in patients with CHF or in patients with COPD. In both control subjects and patients, FFM was the most significant determinant of O(2)peak. CONCLUSION: Muscle dysfunction is not limited to the lower limbs, but generalized and comparable between patients with CHF and patients with COPD with similar exercise capacity. FFM is a strong predictor of peripheral muscle strength, to a lesser extent of O(2)peak, and not at all of peripheral muscle endurance. PMID- 12740257 TI - Augmentation therapy with alpha1-antitrypsin: patterns of use and adverse events. AB - STUDY OBJECTIVES: To describe patterns of prescribing augmentation therapy, and types and rates of adverse events in the National Heart, Lung, and Blood Institute Registry for Individuals with Severe Deficiency of Alpha(1) Antitrypsin. DESIGN: Observational cohort study with follow-up visits every 6 to 12 months for up to 7 years. MEASUREMENTS: The rate and dosing frequency with which Registry participants were prescribed to receive augmentation therapy by their managing physicians, and the type and frequency of adverse events, classified in two ways: severity of self-reported symptoms, and actions taken as a consequence of the symptom. RESULTS: Over the course of Registry follow-up, 66% (n = 747) of the participants received augmentation therapy at some time. In keeping with recommendations made in the 1989 American Thoracic Society (ATS) statement, 75% of participants with airflow obstruction at first visit (defined as FEV(1) < 80% predicted) received augmentation therapy within 3 years, though some participants with FEV(1) > or = 80% predicted (14%) also received augmentation therapy. Among those with COPD for whom augmentation therapy was not prescribed, financial constraints were the reported cause in 30%. Observed patterns also varied from approved practice, in that dosing frequencies other than the US Food and Drug Administration-approved, once-weekly regimen were frequently prescribed. The overall rate of reported adverse events was 0.02 per patient-month, with 83% of participants reporting no events. This overall rate was composed of 16% considered mild events, 76% moderate events, and 9% severe events. CONCLUSIONS: We conclude that augmentation therapy was generally well tolerated and, consistent with ATS guidelines, physicians generally did not prescribe augmentation therapy for subjects with FEV(1) > or = 80% predicted. However, the large percentage of subjects with FEV(1) <80% predicted not receiving augmentation therapy and the frequent use of 2- to 3-week or monthly dosing reflects variation of practice from suggested treatment guidelines. PMID- 12740258 TI - A longitudinal study of alpha1-antitrypsin phenotypes and decline in FEV1 in a community population. AB - BACKGROUND: It is well-known that the homozygous deficiency of alpha(1) antitrypsin, phenotype PiZZ, is associated with an increased risk of COPD. However, studies evaluating the association between the heterozygous forms of the alpha(1)-antitrypsin phenotype PiMZ and rapid decline in lung function, both in patient and community populations, have yielded conflicting results. STUDY OBJECTIVE: To assess the relationship between alpha(1)-antitrypsin phenotypes and decline in FEV(1) values of 2,016 adult subjects in a community population in Tucson, AZ. DESIGN AND METHODS: Prospective cohort study. Standardized questionnaires and lung function measurements were administered 1.5 to 2 years apart during 12 surveys. RESULTS: The frequency distribution for PiMM, PiMS, and PiMZ phenotypes did not differ significantly by physician-confirmed diagnoses of emphysema, chronic bronchitis, or asthma. There was no statistically significant difference in mean FEV(1) slope values between PiMM, PiMS, and PiMZ phenotypes ( 22.5, -21, and -7 mL per year, respectively). After controlling for smoking and other potential confounders, the FEV(1) slope was associated with an initial FEV(1) level and age for the initial questionnaire but not with the different phenotypes. Selecting cutoff values, we identified rapidly declining and nondeclining subgroups, based on the percent predicted changes in FEV(1). They also were not associated with alpha(1)-antitrypsin phenotypes. CONCLUSIONS: We conclude that the data from this longitudinal community study suggest that having the PiMZ phenotype is not a significant risk factor for an accelerated decline in FEV(1). PMID- 12740259 TI - Long-term treatment benefits with tiotropium in COPD patients with and without short-term bronchodilator responses. AB - OBJECTIVES: To determine whether long-term symptomatic improvement occurs in COPD patients with maintenance bronchodilator therapy despite a nonsignificant short term improvement in FEV(1) following bronchodilator inhalation obtained at a single time point. METHODS: Data obtained during two identical 1-year, placebo controlled trials of tiotropium, 18 micro g once daily, were analyzed retrospectively to determine the associations of long-term improvements in lung function and patient health status with short-term improvements in FEV(1), as measured on the first day of treatment. Based on the presence or absence of a short-term improvement in FEV(1) of > or = 12% and > or = 200 mL, respectively, patients who had been treated with tiotropium were characterized as being responsive to tiotropium (TIO-R) or poorly responsive to tiotropium (TIO-PR). RESULTS: Baseline characteristics were similar other than baseline FEV(1), which was higher in the TIO-R group than in both the TIO-PR and placebo groups (p < 0.05). Baseline FEV(1) was 1.08 L in the TIO-R group (n = 263), 0.95 L in the TIO PR (n = 255), and 0.99 L in the placebo group (n = 328). The mean (+/- SD) morning predose FEV(1) at 1 year significantly (p < 0.001) improved in patients in both of the tiotropium treatment subgroups (TIO-R group, 212 +/- 17 mL; TIO-PR group, 94 +/- 17 mL) relative to those treated with placebo. Statistically significant improvements in both tiotropium-treated groups also were noted over 1 year for dyspnea (p < 0.001), as assessed by the transition dyspnea index (TDI) [TIO-R group, 1.36 +/- 0.23 L; TIO-PR group, 0.86 +/- 0.23 L] relative to the placebo group. Patient health status assessed by the St. George Respiratory Questionnaire (SGRQ) showed statistically significant improvements over placebo for the TIO-R and TIO-PR groups (-3.96 +/- 0.99 and -3.05 +/- 1.00 L, respectively; p < 0.005). There was a significant correlation of the first-dose short-term FEV(1) response to the end-of-trial trough response (r = 0.43), but there was only a weak correlation to TDI focal score (r = 0.17) or SGRQ total score (r= -0.12). CONCLUSIONS: Tiotropium was effective in the treatment of patients with COPD, irrespective of the presence or absence of a short-term response on the first day of treatment. The short-term bronchodilator response should not be used as a definitive criterion for prescribing long-term treatment with inhaled bronchodilators. PMID- 12740260 TI - Comparison of acetazolamide and medroxyprogesterone as respiratory stimulants in hypercapnic patients with COPD. AB - BACKGROUND: Acetazolamide and medroxyprogesterone acetate (MPA) are two respiratory stimulants that can be used in patients with stable hypercapnic COPD. DESIGN AND METHODS: The effects of acetazolamide, 250 mg bid, and MPA, 30 mg bid, on daytime and nighttime blood gas values and the influences on the hypercapnic and hypoxic ventilatory and mouth occlusion pressure (P(0.1)) at 100 ms response were studied in a crossover design in 12 hypercapnic patients with stable COPD (FEV(1), 33 +/- 4% predicted [mean +/- SEM]). RESULTS: Daytime PaCO(2) decreased from 47.3 +/- 0.8 mm Hg (placebo) to 42.0 +/- 1.5 mm Hg during acetazolamide treatment (p < 0.05) and to 42.8 +/- 1.5 mm Hg during MPA treatment (p < 0.05). Daytime PaO(2) improved with acetazolamide from 65.2 +/- 2.3 to 75.0 +/- 3.0 mm Hg (p < 0.05), whereas no significant changes were seen with MPA. Mean nocturnal end-tidal carbon dioxide tension decreased with both treatments, from 42.0 +/- 2.3 to 35.3 +/- 2.3 mm Hg with acetazolamide (p < 0.05) and to 34.5 +/- 0.8 mm Hg with MPA (p < 0.05). The percentage of time that the nocturnal arterial oxygen saturation was < 90% was reduced significantly with acetazolamide, from 34.9 +/- 10.7% to 16.3 +/- 7.5% (p < 0.05). Mean nocturnal saturation did not change with MPA. Resting minute ventilation increased significantly only with MPA from 9.6 +/ 0.7 to 10.8 +/- 0.8 L/min (p < 0.05). The slope of the hypercapnic ventilatory response did not change during acetazolamide and MPA therapy. The hypoxic ventilatory response increased from - 0.2 +/- 0.05 to - 0.4 +/- 0.1 L/min/% during acetazolamide (p < 0.05) and to - 0.3 +/- 0.1 L/min/% during MPA (p < 0.05). The hypoxic P(0.1) response improved with acetazolamide treatment from - 0.05 +/- 0.008 to - 0.15 +/- 0.02 mm Hg/% (p < 0.05). CONCLUSIONS: This study shows that acetazolamide and MPA both have favorable effects on daytime and nighttime blood gas parameters in ventilatory-limited patients with stable COPD. However, the use of acetazolamide is preferred because of its extra effect on nocturnal saturation. PMID- 12740261 TI - Prognostic value of nutritional depletion in patients with COPD treated by long term oxygen therapy: data from the ANTADIR observatory. AB - BACKGROUND: An association between weight depletion and mortality has been demonstrated in patients with COPD, but the prognostic influence of malnutrition has not been evaluated in patients with the most severe COPD treated with home long-term oxygen therapy (LTOT). STUDY OBJECTIVE: To analyze the prognostic value of nutritional depletion in patients with COPD receiving LTOT with respect to survival and hospitalization rate. DESIGN: Analysis of a national database (Observatory of Association Nationale pour le Traitement a Domicile de l'Insuffisance Respiratoire Chronique [ANTADIR]). SETTING: The national nonprofit network for home treatment of patients with chronic respiratory insufficiency (ANTADIR) founded in France in the 1980s. PATIENTS: A total of 4,088 patients with a diagnosis of chronic bronchitis or emphysema, FEV(1)/vital capacity ratio < 60%, PaO(2) < 8 kPa, and treatment with LTOT between 1984 and 1993. MEASUREMENTS AND RESULTS: The prevalence of malnutrition, as defined by a body mass index (BMI) < 20, was 23% in men and 30% in women. BMI was significantly correlated with FEV(1) and FEV(1)/VC. The mean follow-up duration was 7.5 years. The 5-year survival rates were 24%, 34%, 44%, and 59%, respectively, for patients with BMIs < 20, 20 to 24, 25 to 29, and > or = 30. Multivariate analysis using the Cox model demonstrated that the effect of BMI on survival was independent of age, FEV(1), PaO(2), and sex. Lower BMI was the most powerful predictor of duration and rate of hospitalization, independently of blood gas levels and respiratory function. The mean (+/- SD) annual time spent in the hospital was 29.6 +/- 40.4 days for patients with a BMI < 20 vs 17.5 +/- 30.1 days for patients with a BMI > 30. CONCLUSION: This study showed that nutritional depletion is an independent risk factor for mortality and hospitalization in patients with COPD receiving LTOT. The best prognosis was observed in overweight and obese patients. PMID- 12740262 TI - Measurement of nasal potential difference in mild asthmatics. AB - STUDY OBJECTIVE: To determine whether ion transport or barrier function across the nasal epithelium are altered in asthmatics. DESIGN: In this pilot study, the nasal potential difference (PD) was measured using the technique established by Knowles and colleagues. A flowing agar bridge made electrical contact with the surface of the nasal epithelium along the floor of the nose. This bridge was referenced to a cutaneous electrode to determine the PD across the nasal epithelium. Changes in nasal PD in response to amiloride, chloride-free medium, and chloride-free medium containing isoproterenol were measured, and responses of asthmatics and healthy control subjects were compared. PARTICIPANTS: Measurements were made in eight adult nonasthmatic subjects and 6 adult asthmatic subjects. All asthmatics had mild intermittent asthma. MEASUREMENTS AND RESULTS: Continuous measurements of nasal PD were obtained while the nasal surface was perfused consecutively with saline solution (NaCl-containing solution), saline solution plus 100 micro mol/L amiloride, chloride-free solution plus amiloride, and chloride-free solution with amiloride plus 10 micro mol/L isoproterenol (a beta adrenergic agonist). No significant differences in baseline PD or change in PD in response to changes in perfusate were found between the two groups. CONCLUSIONS: Our results suggest that ion transport and barrier function of patients with mild asthma are normal. Therefore, by contrast to cystic fibrosis, changes in salt and water transport across airway epithelium may not contribute to accumulation of mucous secretions in asthma. PMID- 12740263 TI - Cigarette smoking among asthmatic adults presenting to 64 emergency departments. AB - STUDY OBJECTIVES: The emergency department (ED) is an important focal point for asthmatic individuals with uncontrolled illness. Anecdotally, many adults presenting to the ED with acute asthma are active cigarette smokers. The present study determined the prevalence of cigarette smoking among adults presenting to the ED with acute asthma and identified the factors associated with current smoking status. DESIGN: A prospective cohort study conducted as part of the Multicenter Airway Research Collaboration. PATIENTS: A structured interview was performed in 1,847 patients, ages 18 to 54 years, who presented to the ED with acute asthma. SETTING: Sixty-four EDs in 21 US states and 4 Canadian provinces. RESULTS: Thirty-five percent of the enrolled asthmatic patients were current smokers with a median of 10 pack-years (interquartile range, 4 to 20 pack-years), while 23% were former smokers, and 42% were never-smokers. Current smokers comprised 33% of asthmatic patients aged 18 to 29 years, 40% for ages 30 to 39 years, and 33% for ages 40 to 54 (p < 0.001). In a multivariate analysis, the factors independently associated with current smoking status (p < 0.05) were as follows: age 30 to 39 years; white race/ethnicity; non-high school graduate; lower household income; lack of private insurance; no recent inhaled steroid usage; and no history of systemic steroid usage. Although 50% of current smokers admitted that smoking worsens their asthma symptoms, only 4% stated that smoking was responsible for their current exacerbation. CONCLUSIONS: Although cigarette smoke is generally recognized as a respiratory irritant, cigarette smoking is common among adults presenting to the ED with acute asthma. The ED visit may provide an opportunity for patients to be targeted for smoking cessation efforts. PMID- 12740264 TI - Budesonide and formoterol in a single inhaler improves asthma control compared with increasing the dose of corticosteroid in adults with mild-to-moderate asthma. AB - BACKGROUND: We evaluated the efficacy and safety of low-dose budesonide/formoterol, 80 micro g/4.5 micro g, bid in a single inhaler (Symbicort Turbuhaler; AstraZeneca; Lund, Sweden) compared with an increased dose of budesonide, 200 micro g bid, in adult patients with mild-to-moderate asthma not fully controlled on low doses of inhaled corticosteroid alone. METHODS: All patients received budesonide, 100 micro g bid, during a 2-week run-in period. At the end of the run-in phase, 467 patients with a mean FEV(1) of 82% predicted received 12 weeks of treatment with budesonide/formoterol in a single inhaler or budesonide alone in a higher dose. Patients kept daily records of their morning and evening peak expiratory flow (PEF), nighttime and daytime symptom scores, and use of reliever medication. RESULTS: The increase in mean morning PEF-the primary efficacy measure-was significantly higher for budesonide/formoterol compared with budesonide alone (16.5 L/min vs 7.3 L/min, p = 0.002). Similarly, evening PEF was significantly greater in the budesonide/formoterol group (p < 0.001). In addition, the percentage of symptom-free days and asthma-control days (p = 0.007 and p = 0.002, respectively) were significantly improved in the budesonide/formoterol group. Budesonide/formoterol decreased the relative risk of an asthma exacerbation by 26% (p = 0.02) compared with budesonide alone. Adverse events were comparable between the two treatment groups. CONCLUSION: This study shows that in adult patients whose mild-to-moderate asthma is not fully controlled on low doses of inhaled corticosteroids, single-inhaler therapy with budesonide and formoterol provides greater improvements in asthma control than increasing the maintenance dose of inhaled corticosteroid. PMID- 12740265 TI - Left ventricular diastolic function in patients with advanced cystic fibrosis. AB - OBJECTIVES: To assess left ventricular systolic and diastolic function in adult patients with cystic fibrosis using radionuclide ventriculography. BACKGROUND: Although myocardial fibrosis has been described in autopsy specimens of patients with cystic fibrosis, the possibility that myocardial dysfunction may occur during life in adult patients with cystic fibrosis has not been explored. METHODS: To assess the possibility of cardiac dysfunction occurring in cystic fibrosis, we studied 40 patients with advanced cystic fibrosis with first-pass radionuclide ventriculography and compared them to 9 patients with advanced bronchiectasis and 18 normal control subjects. RESULTS: Indexes of right ventricular systolic function were similarly impaired in patients with cystic fibrosis and patients with bronchiectasis. Left ventricular ejection fraction of patients with cystic fibrosis, patients with bronchiectasis, and normal control subjects did not differ. Fractional left ventricular filling at 50% of diastole, an index of diastolic function, was significantly lower in patients with cystic fibrosis (54 +/- 13%, mean +/- SD) in comparison to patients with bronchiectasis (66 +/- 4%, p = 0.009) or normal control subjects (69 +/- 14, p = 0.0002). The contribution of atrial systole to total diastolic left ventricular filling was greater in patients with cystic fibrosis (38 +/- 18%) than in patients with bronchiectasis (21 +/- 4%, p = 0.01) or normal control subjects (25 +/- 12%, p = 0.01). CONCLUSIONS: Patients with advanced cystic fibrosis demonstrate impaired left ventricular distensibility when compared to normal control subjects and patients with bronchiectasis. Patients with cystic fibrosis may be at risk of heart failure due to right ventricular dysfunction or left ventricular diastolic dysfunction. PMID- 12740266 TI - Susceptibility testing of Pseudomonas aeruginosa isolates and clinical response to parenteral antibiotic administration: lack of association in cystic fibrosis. AB - STUDY OBJECTIVE: To determine the relationship between the antibiotic susceptibility of Pseudomonas aeruginosa isolated from the sputum of patients with cystic fibrosis (CF) and the patient's response to parenteral antibiotic administration, we performed a retrospective analysis using data from patients in the placebo arm of a phase 3 trial of tobramycin solution for inhalation. All patients were chronically infected with P aeruginosa. Seventy-seven of the 262 patients receiving placebo experienced a pulmonary exacerbation during the trial for which they received therapy with IV tobramycin and ceftazidime. The susceptibility of the P aeruginosa isolates to ceftazidime and tobramycin was determined at trial enrollment by broth microdilution. DESIGN: The clinical response to combination antibiotic therapy was assessed by analyzing differences in spirometry before and after antibiotic administration. The FEV(1) percent predicted at the first visit after the conclusion of antibiotic administration was compared to the FEV(1) percent predicted prior to antibiotic therapy. The results were analyzed both descriptively and by regression analyses. RESULTS: The conditions of 54 patients improved, and those of 9 patients worsened, and in 14 patients there was no change in FEV(1) with antibiotic administration. No correlation was observed between the susceptibility of P aeruginosa to tobramycin or ceftazidime and clinical response. Only the three following variables were observed to significantly correlate with FEV(1) after antibiotic treatment on regression analysis: FEV(1) prior to treatment (p < 0.0001); number of days elapsed between the previous FEV(1) measurement and the initiation of IV antibiotic therapy (p < 0.002); and the number of days elapsed between the determination of the minimum inhibitory concentration and the initiation of IV therapy (p < 0.03). No significant trends were observed between the antibiotic susceptibility of P aeruginosa isolates and treatment outcomes. CONCLUSION: While lack of statistical significance for a trend between bacterial susceptibilities and the response to parenteral antibiotic administration does not mean that no such trend exists, the precision of the confidence intervals allows us to conclude that even if isolate antibiotic susceptibilities affect outcome, the impact would be small and not clinically relevant. PMID- 12740267 TI - Impact of initial antibiotic choice on clinical outcomes in community-acquired pneumonia: analysis of a hospital claims-made database. AB - BACKGROUND: Much controversy exists regarding the initial choice of antibiotics and selected outcomes for patients with community-acquired pneumonia (CAP). METHODS: The investigators analyzed a hospital claims-made database to assess the impact of initial antibiotic choice on 30-day mortality, total hospital costs, and hospital length of stay (LOS). Fine risk groups allowed for stratification for variations in the severity of illness. Patients were divided into five monotherapy groups (ie, ceftriaxone, "other" cephalosporins, fluoroquinolones, macrolides, or penicillins) and four groups that received dual therapy (ie, the agents listed above, except macrolides) plus macrolides. Patients also were stratified by age (ie, > 65 years of age and < 65 years of age). Severely ill patients were excluded. RESULTS: Overall, 44,814 persons met the criteria for inclusion. Among monotherapy patients, those who received macrolides had the least mortality but were the least ill. Patients who received dual therapy generally had shorter LOSs, lower total hospital charges, and decreased mortality compared with those who received monotherapy. Differences among dual-therapy regimens regarding outcomes studies were noted. Patients who were < 65 years of age had lower mortality rates, shorter LOSs, and lower hospital charges than did the more elderly patients. Within this group, those who received dual therapy had better outcomes than those who received monotherapy. CONCLUSIONS: We confirmed the value of dual therapy employing macrolides as a second agent in decreasing mortality from CAP, and we provided similar data regarding shorter LOSs and lower hospital charges. This appears to hold for a younger population. Differences among dual-therapy regimens (all employing macrolides) appear to exist and may be clinically relevant. PMID- 12740268 TI - Community-acquired pneumonia in southeast Asia: the microbial differences between ambulatory and hospitalized patients. AB - STUDY OBJECTIVES: To determine microbial agents causing community-acquired pneumonia (CAP) in Southeast Asia. DESIGN: A prospective study. SETTING: Three general hospitals in Thailand. PATIENTS: Two hundred forty-five adult patients fulfilling the clinical criteria of CAP from September 1998 to April 2001. INTERVENTIONS: Investigations included sputum Gram stain and culture, blood culture, pleural fluid culture (if presented), urine antigen for Legionella pneumophila and Streptococcus pneumoniae, and serology for Mycoplasma pneumoniae, Chlamydia pneumoniae, and L pneumophila. RESULTS: There were 98 outpatients and 147 hospitalized patients included in the study, and an organism was identified in 74 of 98 outpatients (75.5%) and 105 of 147 of the hospitalized patients (71.4%). C pneumoniae (36.7%), M pneumoniae (29.6%), and S pneumoniae (13.3%) were the most frequent causative pathogens found in outpatients, while S pneumoniae (22.4%) and C pneumoniae (16.3%) were the most common in hospitalized patients. There was a significantly higher incidence of C pneumoniae (36.7% vs 16.3%, respectively; p < 0.001) and M pneumoniae (29.6% vs 6.8%; p < 0.001, respectively) in the outpatients than in the hospitalized patients. The incidence of S pneumoniae, L pneumophila, and mixed infections was not different between the groups. Mixed infections were presented in 13 of 98 outpatients (13.3%) and 9 of 147 hospitalized patients (6.1%), with C pneumoniae being the most frequent coinfecting pathogen. CONCLUSIONS: The data indicate that the core organisms causing CAP in Southeast Asia are not different from those in the Western countries. The guidelines for the treatment of patients with CAP, therefore, should be the same. PMID- 12740269 TI - Vascular endothelial growth factor gene polymorphisms in Japanese patients with sarcoidosis. AB - OBJECTIVES: Vascular endothelial growth factor (VEGF) promotes angiogenesis, mediates vascular permeability, and activates and recruits monocytes. VEGF is produced in activated alveolar macrophages, in epithelioid cells, and in multinuclear giant cells of pulmonary sarcoid granulomas. Recent reports have shown that a polymorphism at - 627 of the VEGF gene is related to VEGF protein production, and a polymorphism at + 813 is associated with VEGF plasma levels. We investigated the roles of such polymorphisms in the development and extent of sarcoidosis. METHODS: We examined polymorphisms of the VEGF gene in 103 Japanese patients with sarcoidosis and 146 healthy Japanese control subjects. The position - 627 polymorphism was determined using the TaqMan (TaqMan Laboratory, University of Pittsburgh Cancer Institute; Pittsburgh, PA) polymerase chain reaction (PCR) method. For genotyping of the position + 813 polymorphism, the PCR restriction fragment length polymorphism method was performed. RESULTS: As for + 813 genotypes, the less-common genotypes CT and TT were found more often in control subjects than in patients (odds ratio, 0.490; 95% confidence interval, 0.276 to 0.868). A significant increase in the frequency of the T allele (p = 0.005, Pc = 0.020 after Bonferroni correction) was observed in control subjects. As for - 627 genotypes, the mean value of the FEV(1)/FVC percentage in GG type was lower than that in CC or CG type, however, the other clinical findings did not suggest airway diseases in the GG type. CONCLUSIONS: We suggest that in VEGF gene polymorphisms the T allele at + 813 may decrease susceptibility to sarcoidosis. PMID- 12740270 TI - Occupational risk factors for sarcoidosis in African-American siblings. AB - OBJECTIVES: To determine whether certain occupations and occupationally related exposures were associated with a history of sarcoidosis in African-American siblings. METHODS: We collected occupational data from 921 African Americans in 273 sibships that had been identified through a sarcoidosis case. Among the 648 siblings of sarcoidosis index cases enrolled, 30 (4.6%) also had a history of sarcoidosis. A detailed job history was obtained for any job held for > or = 6 months throughout the subject's life. RESULTS: Having a usual occupation in education (odds ratio [OR], 2.18; 95% confidence interval [CI], 1.07 to 4.44), in metal machining (OR, 7.47; 95% CI, 1.19 to 47.06), and ever working in metalworking, not elsewhere classified (OR, 2.05; 95% CI, 1.14 to 3.70) were associated with increased sarcoidosis risk. Occupations ever held in the transportation services industry (OR, 12.71; 95% CI, 1.32 to 122.56) and usual occupations in the retail trade industry (OR, 0.49; 95% CI, 0.27 to 0.88) also were associated with sarcoidosis risk. Specific occupational exposures that were associated with sarcoidosis included titanium (OR, 3.15; 95% CI, 1.02 to 9.68) and vegetable dust (OR, 1.82; 95% CI, 1.01 to 3.27), and indoor exposure to high humidity (OR, 1.51; 95% CI, 1.13 to 2.02), water damage (OR, 1.50; 95% CI, 1.11 to 2.03), or musty odors (OR, 1.78; 95% CI, 1.32 to 2.40) for > 1 year. CONCLUSION: Individuals who work in occupations with potential metal exposures or in workplaces with high humidity may be at an increased risk for sarcoidosis, but the complexity of occupationally related exposures makes it difficult to identify specific agents by using job titles as a surrogate for exposure. A more detailed exposure assessment of such jobs, along with the incorporation of genetic risk factors, should help to uncover the complex etiology of sarcoidosis. PMID- 12740271 TI - Relationship of systolic BP to obstructive sleep apnea in patients with heart failure. AB - STUDY OBJECTIVES: Obstructive sleep apnea (OSA) is an independent risk factor for hypertension in the general population. Hypertension is, in turn, an important risk factor for the development and progression of congestive heart failure (CHF). Our objective was to determine whether OSA would be associated with elevated daytime BP in medically treated patients with CHF. DESIGN: Cross sectional study. SETTING: Tertiary care, university-affiliated sleep disorders and heart failure clinics. PATIENTS: Three hundred one consecutive patients with CHF. MEASUREMENTS AND RESULTS: We measured daytime BP and performed overnight sleep studies to assess for the presence of OSA. Among these patients, OSA was present in 121 patients (40%) and their systolic BP was significantly higher than in patients without OSA. Patients with OSA were 2.89 times (95% confidence interval, 1.25 to 6.73) more likely to have systolic hypertension (ie, BP > or = 140 mm Hg) than those without OSA after controlling for other risk factors, including obesity. The degree of systolic BP elevation was directly related to the frequency of obstructive apneas and hypopneas. CONCLUSIONS: In medically treated patients with CHF, daytime systolic BP and the prevalence of systolic hypertension are significantly increased in patients with OSA, compared to those without OSA, independent of other potentially confounding factors. OSA may therefore have contributed to the presence of systolic hypertension in some of these patients. PMID- 12740272 TI - Gender differences in sleep apnea: the role of neck circumference. AB - STUDY OBJECTIVES: To determine whether differences in sleep apnea severity between men and women referred to a sleep clinic are related to the differences in neck circumference (NC). STUDY DESIGN: Case series. SETTING: University hospital sleep disorders clinic. PARTICIPANTS: A total of 3,942 patients (2,753 men and 1,189 women) referred to the sleep clinic. MEASUREMENTS AND RESULTS: All patients underwent nocturnal polysomnography. NC was used as a surrogate measure of upper airway obesity. We found that sleep apnea, defined an the apnea/hypopnea index (AHI) > 10/h, was significantly more frequent (60% vs 32%, chi(2) < 0.0001) and severe (mean +/- SE, 25 +/- 26/h vs 12 +/- 19/h, p < 0.0001) in men than in women. Men had significantly larger NC than women, but the difference became much less pronounced when we normalized NC to body height (0.24 +/- 0.02 vs 0.23 +/- 0.03, p < 0.0001). Men had significantly higher AHI than women even after controlling for age, body mass index (BMI), and neck/height ratio (NHR); analysis of covariance showed that mean AHI was 24.4 +/- 0.4 in men vs 14.8 +/- 0.7 in women (p < 0.0001). This difference persisted even when we matched men and women for NHR and BMI. Finally, multiple regression analysis revealed the following: (1) NHR was the most significant predictor of AHI, accounting for 19% of the variability; and (2) the slope of AHI vs NHR was significantly higher in men than in women. CONCLUSIONS: We conclude the following: (1) the frequency and severity of sleep apnea in the sleep clinic population is greater in men than women, and (2) factors other than NC, age, and BMI must contribute to these gender differences. PMID- 12740273 TI - Cardiorespiratory effects of added dead space in patients with heart failure and central sleep apnea. AB - BACKGROUND: Inhaled CO(2) has been shown to stabilize the breathing pattern of patients with central sleep apnea (CSA) with and without congestive heart failure (CHF). Added dead space (DS) as a form of supplemental CO(2) was effective in eliminating idiopathic CSA. The efficacy and safety of DS has not yet been evaluated in patients with CHF and CSA. METHODS: We examined the respiratory and cardiovascular effects of added DS in eight patients with CHF and CSA. The DS consisted of a facemask attached to a cylinder of adjustable volume. During wakefulness, the cardiorespiratory response to 200 to 600 mL of DS was tested. Cardiac output and stroke volume were measured using echocardiography with and without DS. During the nocturnal study, patients slept with and without DS, alternating at approximately 1-h intervals. RESULTS: Values are expressed as the mean +/- SE. The wakefulness study revealed a plateau in the partial pressure of end-tidal CO(2) (PETCO(2)) and the partial pressure of end-tidal O(2) between DS amounts of 400 and 600 mL. The mean stroke volume index (33 +/- 7 vs 34 +/- 7 mL/m(2), respectively) and the mean cardiac index (1.9 +/- 0.3 vs 1.9 +/- 0.4 L/min/m(2), respectively) were not affected by DS. Neither heart rate nor BP showed a significant change in response to DS of < or = 600 mL. During sleep, DS increased the PETCO(2) (40.7 +/- 2.7 vs 38.9 +/- 2.6 mm Hg, respectively; p < 0.05), reduced apnea (1 +/- 1 vs 29 +/- 7 episodes per hour, respectively; p < 0.01) and arousal (21 +/- 8 vs 30 +/- 8 arousals per hour, respectively; p < 0.05), increased the mean arterial oxygen saturation (SaO(2)) [94.4 +/- 1.0% vs 93.5 +/- 1.1%, respectively; p < 0.01), and reduced SaO(2) oscillations (DeltaSaO(2) from maximum to minimum, 1.8 +/- 0.4% vs 5.5 +/- 0.9%, respectively; p < 0.01). CONCLUSION: DS stabilized CSA and improved sleep quality in patients with CHF without significant acute adverse effects on the cardiovascular function. PMID- 12740274 TI - BP and arterial distensibility in children with primary snoring. AB - STUDY OBJECTIVE: While previous studies have suggested an association between obstructive sleep apnea and cardiovascular complications, the effects of primary snoring in children on daytime systemic BP and arterial distensibility remain unknown. DESIGN AND PATIENTS: To determine the effects of primary snoring on BP and peripheral conduit artery distensibility, 30 children with primary snoring were studied at an age of 9.5 +/- 2.8 years (mean +/- SD). Systemic BP was measured using an automated device, while brachioradial arterial distensibility was assessed by measuring pulse wave velocity (PWV), which is inversely related to the square root of distensibility. The results were compared to those of 30 healthy control subjects matched for age, sex, and body size. RESULTS: As compared to control subjects, children with primary snoring had significantly higher systolic BP (112 +/- 10 mm Hg vs 105 +/- 8 mm Hg, p = 0.001), diastolic BP (60 +/- 7 mm Hg vs 53 +/- 9 mm Hg, p = 0.004), and mean BP (81 +/- 7 mm Hg vs 71 +/- 8 mm Hg, p < 0.001). Likewise, those with primary snoring had significantly higher PWV (9.7 +/- 1.6 m/s vs 7.9 +/- 2.0 m/s, p = 0.001). Multiple regression identified age, body mass index (BMI), and primary snoring as significant determinants of systemic BP; however, primary snoring is the only significant determinant of PWV. Regardless of the BMI, systemic BP and PWV remained significantly higher in children with primary snoring. CONCLUSION: Children with primary snoring have increased daytime systemic BP and reduced arterial distensibility, which may jeopardize long-term cardiovascular health. PMID- 12740275 TI - Utility of oxygen saturation and heart rate spectral analysis obtained from pulse oximetric recordings in the diagnosis of sleep apnea syndrome. AB - OBJECTIVES: We prospectively evaluate the spectral characteristics of nocturnal arterial oxygen saturation (SaO(2)) and heart rate variability obtained from pulse oximetric recording as a diagnostic test for obstructive sleep apnea (OSA). SUBJECTS AND MEASUREMENTS: Three hundred referred outpatients with symptoms compatible with the diagnosis of OSA were studied using nocturnal pulse oximetric recording performed simultaneously with polysomnography. Power spectral analysis of SaO(2) and heart rate data were analyzed using fast Fourier transformation of a Hamming-windowed signal. DESIGN AND RESULTS: Recording test results were classified as abnormal (suspicion of OSA) if the periodogram showed a peak in the period 30 to 70 s in either of the signals. A normal test result was defined as the absence of this peak in the periodogram in both signals. Two independent observers performed a single-blind evaluation. The total area of the periodogram (STOT), the ratio of the area enclosed in the periodogram within the period 30 to 70 s (S(30-70)), the ratio of the area enclosed in the periodogram within the period 30 to 70 s with respect to the total area of the periodogram (S), and the peak amplitude of the periodogram in the period 30 to 70 s (PA) were measured in both signals. The presence of a peak in the periodogram in either of the signals has a sensitivity of 94%, a specificity of 82%, a positive predictive value of 87%, and a negative predictive value of 92% with respect to the OSA diagnosis. The patients in the OSA group had higher values for STOT, S(30-70), S, and PA than the group without OSA. CONCLUSIONS: SaO(2) and heart rate spectral analysis obtained by nocturnal pulse oximetry as well as the identification of a peak within 30 to 70 s in either signal could be useful as a diagnostic technique for patient with OSA. PMID- 12740276 TI - Pulmonary hypertension in patients with end-stage renal disease. AB - BACKGROUND: The aims of this study were to evaluate the incidence of unexplained pulmonary hypertension (PH) among patients with end-stage renal disease (ESRD) and to suggest possible etiologic factors. METHODS: The incidence of PH was prospectively estimated by Doppler echocardiography in 58 patients with ESRD receiving long-term hemodialysis via arteriovenous access, and in control groups of 5 patients receiving peritoneal dialysis (PD) and 12 predialysis patients without a known other cause to suggest the presence of PH. Clinical variables were compared between patients with and without PH receiving hemodialysis. Changes in pulmonary artery pressure (PAP) values before and after onset of hemodialysis via arteriovenous access, arteriovenous access compression, and successful kidney transplantation were recorded. RESULTS: PH > 35 mm Hg was found in 39.7% of patients receiving hemodialysis (mean +/- SD, 44 +/- 7 mm Hg; range, 37 to 65 mm Hg), in none of the patients receiving PD, and in 1 of 12 predialysis patients. Patients with PH receiving hemodialysis had a significantly higher cardiac output (6.9 L/min vs 5.5 L/min, p = 0.017). PH developed in four of six patients with normal PAP after onset of hemodialysis therapy via arteriovenous access. One-minute arteriovenous access compression in four patients decreased the mean systolic PAP from 52 +/- 7 to 41 +/- 4 mm Hg (p = 0.024). PH normalized in four of five patients receiving hemodialysis following kidney transplantation. Kaplan-Meier survival analysis according to PAP values revealed significant survival differences (p < 0.024). CONCLUSIONS: This study demonstrates a surprisingly high incidence of PH among patients with ESRD receiving long-term hemodialysis with surgical arteriovenous access. Both ESRD and long-term hemodialysis via arteriovenous access may be involved in the pathogenesis of PH by affecting pulmonary vascular resistance and cardiac output. PMID- 12740277 TI - Effect of orally active prostacyclin analogue on survival in patients with chronic thromboembolic pulmonary hypertension without major vessel obstruction. AB - OBJECTIVES: This study investigated whether treatment with beraprost sodium (BPS), an orally active prostacyclin analog, improves hemodynamics and survival in patients with peripheral-vessel chronic thromboembolic pulmonary hypertension (CTEPH), for which there is no surgical option. BACKGROUND: Oral administration of BPS has been shown to improve the hemodynamics and prognosis in patients with primary pulmonary hypertension; however, whether BPS has beneficial effects in CTEPH remains unknown. METHODS: Forty-three patients with peripheral-vessel CTEPH were classified into two groups: patients treated with BPS (BPS group, n = 20) and those without BPS (conventional group, n = 23). Baseline demographic and hemodynamic data did not significantly differ between the two groups. RESULTS: BPS therapy improved New York Heart Association functional class in 10 patients (50%) and significantly decreased total pulmonary resistance from 18 +/- 6 to 15 +/- 8 Wood units (p < 0.05) [mean +/- SD]. Sixteen patients died of cardiopulmonary causes in the conventional group during a mean follow-up period of 58 +/- 45 months. In contrast, only three patients died of cardiopulmonary causes in the BPS group during a mean follow-up period of 44 +/- 30 months. The absence of BPS therapy, elevated total pulmonary resistance, heart rate, and age were independently related to the mortality by Cox proportional hazard analysis. The 1-year, 3-year, and 5-year survival rates for the BPS group were 100%, 85%, and 76%, respectively, compared to 87%, 60%, and 46% in the conventional group. CONCLUSIONS: This preliminary study suggests that oral administration of BPS may improve hemodynamics and survival in patients with peripheral-vessel CTEPH, for which there is no surgical option. PMID- 12740278 TI - Complications of intermittent prone positioning in pediatric patients receiving extracorporeal membrane oxygenation for respiratory failure. AB - STUDY OBJECTIVE: To describe the safety and risks of placing pediatric patients in the prone position during extracorporeal membrane oxygenation (ECMO) for the treatment of respiratory failure. DESIGN: Single-center retrospective cohort study. SETTING: Tertiary pediatric ICUs. PATIENTS: All patients admitted to the pediatric ICU who required ECMO for respiratory failure from 1995 to 2000. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: Medical records for 93 patients representing 95 ECMO runs for treatment of respiratory failure were reviewed. Of these, 63 patients (66%) received intermittent prone positioning. Demographic data and clinical information were recorded. The median age was 12 months, and the median weight was 9.8 kg. There were 962 position changes. Complications surveyed for included bleeding from appliance insertion sites, appliance dislodgment, unplanned extubation, cutaneous pressure ulceration, corneal abrasion, and extreme hemodynamic instability. These complications were noted as to whether they were present prior to the initiation of prone positioning or whether they developed after prone positioning began. Twenty-four percent of patients had bleeding from cannulation sites prior to prone positioning, and 18% of patients had bleeding begin after prone positioning was initiated. Two patients had chest tubes dislodge after prone positioning began, but neither patient had bleeding occur or required reinsertion of the chest tube. There were no unplanned extubations, appliance displacements, development of cutaneous pressure ulcerations, or corneal abrasions associated with prone-positioning maneuvers. No patient had ECMO support removed secondary to the surveyed complications. Eighty-two percent of children who received prone positioning during ECMO for treatment of respiratory failure survived to hospital discharge. CONCLUSION: We found that prone positioning may be used in pediatric ECMO patients without increasing the risk of complications. A multi-institutional, prospective, randomized, controlled study would better evaluate the efficacy of this practice and whether it is associated with a shorter length of ECMO or shorter post-ECMO ventilation and outcome. PMID- 12740279 TI - Percutaneous dilatational tracheostomy in the ICU: optimal organization, low complication rates, and description of a new complication. AB - STUDY OBJECTIVES: To assess short-term and long-term complications of bronchoscopy-guided, percutaneous dilatational tracheostomy (PDT) and surgical tracheostomy (ST) and to report a complication of PDT that has not been described previously. DESIGN: Prospective survey. SETTING: University teaching hospital. PATIENTS: Two hundred eleven critically ill patients in our ICU. INTERVENTIONS: PDT was performed in 174 patients, under bronchoscopic guidance in most cases. ST was performed in 40 patients. RESULTS: No procedure-related fatalities occurred during PDT or ST. The incidence of significant complications (eg, procedure related transfusion of fresh-frozen plasma, RBCs, or platelets, malpositioning or kinking of the tracheal cannula, deterioration of respiratory parameters lasting for > 36 h following the procedure, or stomal infection) in patients undergoing PDT was 4.0% overall and 3.0% when bronchoscopic guidance was used. No cases of paratracheal insertion, pneumothorax, pneumomediastinum, tracheal laceration, or clinically significant tracheal stenosis occurred in patients undergoing PDT. We attribute this low rate of complications to procedural and organizational factors such as bronchoscopic guidance, performance by or supervision of all PDTs by physicians with extensive experience in this procedure, and airway management by physicians who were well-versed in (difficult) airway management. In addition, an ear-nose-throat surgeon participated in the procedure in case conversion of the procedure to an ST should become necessary. We observed a complication that, to our knowledge, has not been reported previously. Five patients developed intermittent respiratory difficulties 2 to 21 days (mean, 8 days) after undergoing PDT. The cause turned out to be the periodic obstruction of the tracheal cannula by hematoma and the swelling of the posterior tracheal wall, which had been caused by intermittent pressure and chafing of the cannula on the tracheal wall. In between the episodes of obstruction, the cannula was open and functioning normally, which made the diagnosis difficult to establish. CONCLUSIONS: Bronchoscopy-assisted PDT is a safe and effective procedure when performed by a team of experienced physicians under controlled circumstances. The intermittent obstruction of the cannula caused by swelling and irritation of the posterior tracheal wall should be considered in patients who develop unexplained paroxysmal respiratory problems some time after undergoing PDT or ST. PMID- 12740280 TI - The utility of chest radiographs following percutaneous dilatational tracheostomy. AB - OBJECTIVE: To determine the usefulness of performing routine postprocedure chest radiographs (CXRs) following percutaneous dilatational tracheostomy (PDT) under bronchoscopic visualization. DESIGN: Retrospective, chart review. SETTING: Medical-surgical ICU of a teaching hospital. METHODS: Sixty patients who underwent PDT under bronchoscopic visualization, for persistent ventilator dependence, in our medical-surgical ICU were studied. We noted the occurrence of any perioperative complications, physician anticipation of postoperative complications, and postprocedure CXR findings. We compared postprocedure CXRs with the last preprocedure CXR. Immediate postoperative complication was defined as a clinical worsening, hemodynamic instability, or a new radiographic finding such as atelectasis, paratracheal placement, pneumothorax, and pneumomediastinum. We determined whether the postprocedure CXRs led to any change in patient management. OBSERVATION: Two patients (3.3%) had postoperative complications detected on their CXRs, one with a pneumomediastinum and the other with a tension pneumothorax. Both procedures were noted to be difficult, with a high physician anticipation of complication. In the remaining 58 patients (96.7%), no postoperative complications were detected on the postprocedure CXRs that necessitated any change in patient management. CONCLUSION: We concluded that routine CXR following PDT that had been performed under bronchoscopic visualization is unnecessary in the absence of clinical deterioration or the anticipation of postoperative complications. PMID- 12740281 TI - Utility of daily routine portable chest radiographs in mechanically ventilated patients in the medical ICU. AB - STUDY OBJECTIVES: To determine whether there is any difference in diagnostic, therapeutic, and outcome efficacy between protocols utilizing daily (ie, routine) chest radiographs (CXRs) and those utilizing clinically indicated (ie, nonroutine) CXRs in mechanically ventilated patients. DESIGN: Prospective, randomized, observational study. SETTING: A 20-bed medical ICU at a university hospital. PATIENTS: Adult patients who had been receiving mechanical ventilation for > 48 h. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: Ninety-four patients who had been intubated for at least 48 h were randomized to receive either routine or nonroutine CXRs and were observed until extubation or death. The percentage of CXRs with new findings was significantly larger in the nonroutine CXR group (53.1%; 120 CXRs) compared to that in the routine CXR group (33.4%; 98 CXRs; odds ratio [OR], 1.59; 95% confidence interval [CI], 1.16 to 2.18; p = 0.004). The number of CXRs with new findings that resulted in interventions was significantly larger in the nonroutine CXR group (26.5%; 60 CXRs) compared to that in the routine CXR group (13.3%; 39 CXRs; OR, 2.0; 95% CI, 1.29 to 3.08; p = 0.002). There was no difference in the mean duration of mechanical ventilation (routine CXR arm, 7.93 days; nonroutine CXR arm, 6.76 days; p = 0.2606), length of ICU stay (routine CXR arm, 11.93 days; nonroutine CXR arm, 9.55 days; p = 0.1936), and total length of hospital stay (routine CXR arm, 19.34 days; nonroutine CXR arm, 16.45 days; p = 0.2199) between the two groups. Although patients in the nonroutine arm of the study received fewer CXRs, adverse outcomes (eg, length of mechanical ventilation, length of hospital stay, length of ICU stay, and mortality) did not increase (p = 0.818). Using the number of CXRs per patient as a surrogate, there was a statistically significant cost savings and decreased radiation exposure in the nonroutine CXR group. CONCLUSIONS: For mechanically ventilated patients, a strategy calling for daily routine CXRs compared to CXRs obtained based on clinical indications alone was not associated with reduced ICU or hospital length of stay or with reduced mortality. PMID- 12740282 TI - Community-acquired bloodstream infection in critically ill adult patients: impact of shock and inappropriate antibiotic therapy on survival. AB - DESIGN: The objectives were to characterize the prognostic factors and evaluate the impact of inappropriate empiric antibiotic treatment and systemic response on the outcome of critically ill patients with community-acquired bloodstream infection (BSI). PATIENTS: A prospective, multicenter, observational study was carried out in 339 patients admitted in 30 ICUs for BSI. RESULTS: Crude mortality was 41.5%. Septic shock was present in 184 patients (55%). The pathogens most frequently associated with septic shock or death were Escherichia coli, Staphylococcus aureus, and Streptococcus pneumoniae, which accounted for approximately half of the deaths. Antibiotic treatment was found to be inappropriate in 14.5% of episodes. Patients in septic shock with inappropriate treatment had a survival rate below 20%. Multivariate analysis identified a significant association between septic shock and four variables: age > or = 60 years (odds ratio [OR], 1.96), previous corticosteroid therapy (OR, 2.58), leukopenia (OR, 2.32), and BSI secondary to intra-abdominal (OR, 2.38) and genitourinary tract (OR, 2.29) infections. The variables that independently predicted death at ICU admission were APACHE (acute physiology and chronic health evaluation) II score > or = 15 (OR, 2.42), development of septic shock (OR, 3.22), and inappropriate empiric antibiotic treatment (OR, 4.11). This last variable was independently associated with an unknown source of sepsis (OR, 2.49). Mortality attributable to inappropriate antibiotic treatment increased with the severity of illness at ICU admission (10.7% for APACHE II score < 15 and 41.8% for APACHE II score > or = 25, p < 0.01). CONCLUSIONS: Inappropriate antimicrobial treatment is the most important influence on outcome in patients admitted to the ICU for community-acquired BSI, particularly in presence of septic shock or high degrees of severity. Initial broad-spectrum therapy should be prescribed to septic patients in whom the source is unknown or in those requiring vasopressors. PMID- 12740283 TI - Time course of expiratory flow limitation in COPD patients during acute respiratory failure requiring mechanical ventilation. AB - STUDY OBJECTIVES: (1) To determine the incidence of expiratory flow limitation (FL) at ICU admission, at the time of extubation, and at ICU discharge in intubated patients with COPD receiving mechanical ventilation for acute respiratory failure (ARF); and (2) to assess the feasibility of inspiratory capacity (IC) as an indication of pulmonary dynamic hyperinflation in this setting. DESIGN: Prospective, observational pilot study with physiologic measurements performed at ICU admission and during the weaning process driven by the clinician. A 60-min T-tube trial was initiated once criteria for weaning were present. The decision to extubate or reventilate patients was made by the clinician at the end of this session. Assessment of failure or success of T-tube trials was performed independently. SETTING: A 25-bed ICU of a tertiary teaching university hospital. PATIENTS: Over a 13-month period, 25 intubated patients with COPD receiving mechanical ventilation for ARF were included. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: At ICU admission, FL assessed by the negative expiratory pressure test was measured under passive ventilatory conditions at the baseline ventilatory settings, on zero end-expiratory pressure, and in a semirecumbent position. During weaning, FL, respiratory pattern, and IC were measured during T-tube trials, before extubation, 1 h after extubation, and at ICU discharge. At ICU admission, 24 of 25 patients presented FL with, on average, 73 +/- 22% of the tidal volume. Ten patients were unavailable for follow-up due to death (n = 6) unplanned extubation (n = 3), or refusal (n = 1), so that only 15 patients completed the whole protocol (all 15 patients were extubated). For these 15 patients, the incidence of FL was 93% at ICU admission, 47% before extubation, and 40% at ICU discharge. IC was significantly greater at ICU discharge than before extubation (36 +/- 11% predicted vs 44 +/- 12% predicted, p < 0.01) and in successful T-tube trials compared with unsuccessful T-tube trials (38 +/- 13% predicted vs 24 +/- 8% predicted, p < 0.01). CONCLUSIONS: The incidence of expiratory FL is high in patients with COPD receiving mechanical ventilation, and is reduced during aggressive therapy when the patient is placed on mechanical ventilatory support and the time that weaning begins during the ICU stay. IC was lower in patients in whom weaning was unsuccessful. Further large scale studies are required to confirm these preliminary results. PMID- 12740284 TI - Matrix metalloproteinases inhibition attenuates tobacco smoke-induced emphysema in Guinea pigs. AB - STUDY OBJECTIVE: To evaluate the effect of CP-471,474 (Pfizer Global Research and Development; Groton, CT), a broad-spectrum inhibitor of matrix metalloproteinases (MMPs) in an experimental model of emphysema. DESIGN: Randomized, double-blinded, controlled experiment. SETTING: Biochemistry and morphology laboratories and animal research facility. METHODS: Guinea pigs were exposed to cigarette smoke over 1 month, 2 months, and 4 months, and half of the animals received CP 471,474. Age-matched guinea pigs exposed to room air were used as control animals. After death, the lungs were lavaged with saline solution, and MMPs in the lavage fluid were determined by zymography and immunoblot. Lungs were fixed for histology, immunohistochemistry, and morphometry. RESULTS: Following a 1 month exposure to tobacco smoke, semiquantitative histologic assessment showed moderate lung inflammation, which progressed in extent and severity and reached a peak at 2 months. CP-471,474 significantly reduced both the extent (p < 0.002) and severity (p < 0.05) of inflammation at 2 months. At 4 months, a spontaneous reduction of the inflammatory response was observed in both treated and untreated animals, and consequently no difference was observed between both. Emphysematous changes, revealed by a significant increase in the average size of alveoli, were detected at 2 months and 4 months of tobacco smoke exposure. The inhibitor significantly decreased the destructive lesions mainly at 2 months (p < 0.0001) and also at 4 months (p < 0.02). Smoking increased MMP-9 and MMP-1 activities as shown by zymography and immunoblot. Immunoreactive MMP-9 was mainly localized in alveolar and bronchiolar epithelial cells, macrophages, and airways smooth-muscle cells. CONCLUSION: These findings support a role for MMPs in the early inflammatory response and in the emphysematous lesions provoked by cigarette smoking. PMID- 12740285 TI - Cigarette smoking increases neutrophil formyl methionyl leucyl phenylalanine receptor numbers. AB - STUDY OBJECTIVES: The purpose of this study was to explore the relationship between cigarette smoking and COPD on the number of formyl methionyl leucyl phenylalanine (FMLP) receptors on peripheral neutrophils. DESIGN AND PARTICIPANTS: Three groups of subjects were studied: subjects with COPD (n = 13), healthy smokers (n = 6), and healthy nonsmokers (n = 6). INTERVENTIONS: Fifty milliliters of venous blood were collected from each subject, and neutrophils were isolated. Neutrophil FMLP receptor numbers were determined by incubating with tritiated FMLP at six doubling concentrations from 1.4 to 45 pmol. Three of the subjects from group 1 (the COPD group) were current smokers, and we elected to analyze these subjects as a separate group. MEASUREMENTS AND RESULTS: The analysis of variance looking at the three factors-FMLP, COPD and smoking-showed significant differences among levels of FMLP (p = 0.0001), as would be expected, and also overall smoking vs nonsmoking (p = 0001) and COPD vs non-COPD (p = 0.02). Within each level of FMLP, there was only one instance of a significant difference between COPD nonsmokers and normal nonsmokers, and no instance of a significant difference between COPD smokers and normal smokers. At five of the six concentrations of tritiated FMLP, smoking was a significant factor. CONCLUSIONS: This study suggests that the overriding influence on peripheral neutrophil FMLP receptor numbers is current smoking rather than the presence of COPD. PMID- 12740286 TI - Continuous tepid blood cardioplegia can preserve coronary endothelium and ameliorate the occurrence of cardiomyocyte apoptosis. AB - OBJECTIVE: In modern cardiac surgery, crystalloid or blood cardioplegic solutions have been used widely for myocardial protection; however, ischemia does occur during protection with intermittent infusion of cold crystalloid or blood cardioplegic solutions. The present study was designed to evaluate the effect of different cardioplegic methods on myocardial apoptosis and coronary endothelial injury after global ischemia, cardiopulmonary bypass (CPB), and reperfusion in anesthetized open-chest dogs. METHODS: The dogs were classified into five groups to identify the injury of myocardium and coronary endothelium: group 1, normothermic CPB without cardiac arrest; group 2, hypothermic CPB with continuous tepid blood cardioplegia, and with cardiac arrest; group 3, hypothermic CPB with intermittent cold blood cardioplegia, and with cardiac arrest; group 4, hypothermic CPB with intermittent cold crystalloid cardioplegia, and with cardiac arrest; and group 5, sham-operated control group. During CPB, cardiac arrest was achieved with different cardioplegia solutions for 60 min, followed by reperfusion for 4 h before the myocardium and coronary arteries were harvested. Coronary arteries were harvested immediately and analyzed by scanning electron microscopy. Cardiomyocytic apoptosis was detected using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling, Western blot, and DNA ladder methods. RESULTS: Regardless of the detection method used, significantly higher percentages of apoptotic cardiomyocytes were found in group 3 and group 4 than in other groups. Expression of caspase-3 correlated with increased apoptosis. Scanning electron microscopy revealed severe endothelial injury of coronary arteries in group 3 and group 4. CONCLUSION: These results point to an important explanation for the difference in cardiac recovery after hypothermic ischemia and arrest with various cardioplegic solutions. PMID- 12740287 TI - An automated approach to quantitative air trapping measurements in mild cystic fibrosis. AB - PURPOSE: To automatically derive the degree of air trapping in mild cystic fibrosis (CF) disease from high-resolution CT (HRCT) data, and to evaluate the discriminating power of the measurement. MATERIALS AND METHODS: The data consist of six pairs of anatomically matched tomographic slices, obtained during breath holding in triggered HRCT acquisitions. The pairs consist of an inspiratory slice, at > or = 95% of slow vital capacity, and an expiratory slice at near residual volume (nRV). The subjects are 25 patients with mild CF and 10 age matched, normal control subjects. SUBJECTS: Lung segmentation is automatic. The limits defining air trapping in the expiratory slices are determined by the distribution of densities in the expanded lung. They are modulated by density changes between expiration and inspiration. Air trapping defects consist of contiguous low-density voxels. The difference between patients and control subjects was evaluated in comparison to pulmonary function test (PFT) results and lung density distribution descriptors (global density descriptors). RESULTS: In mild CF, air trapping does not correlate with global PFT results, except for the ratio of residual volume (RV) to total lung capacity (TLC); however, the size of air trapping defects was the best discriminator between patients and control subjects (p < 0.005). Of PFT results, only RV/TLC reached significance at p < 0.05. The global density descriptors reached near significance in the nRV images only. CONCLUSION: Air trapping defined as defect size and measured in an objective automated manner is a powerful discriminator for mild CF. PMID- 12740288 TI - Diagnosis and treatment of rhinovirus respiratory infections. AB - Acute upper viral respiratory infection (VRI) is the number one cause of illness for which patients seek medical care in the United States. Rhinoviruses, members of the family Picornaviridae, are the causative pathogens in more than half of VRIs, and they are associated with acute exacerbations of respiratory disease, including asthma, sinusitis, otitis media, and COPD. Owing to the lack of commercial availability of rapid and cost-effective laboratory tests to confirm the presence of VRI, the diagnosis is most commonly made empirically, based on patient history and physical examination. Currently, no antiviral agents that are active against picornaviruses are available for clinical use. Antimicrobial agents, frequently prescribed for VRIs, are not active against viruses, and their inappropriate and widespread use has contributed to an increase in antimicrobial resistance among bacteria commonly involved in respiratory tract infections. Several newer antiviral agents are being evaluated for treatment of VRIs. Although a variety of mechanisms and agents have been tested, few have shown significant clinical benefit in human trials. The most advanced antiviral agent in clinical trials is pleconaril, a novel viral capsid-binding inhibitor with potent and highly specific in vitro activity against the majority of serotypes of rhinoviruses and enteroviruses. Clinical trials of pleconaril for the treatment of VRIs have been conducted, and the role of pleconaril in patients with chronic lung disease is being evaluated. PMID- 12740289 TI - Pulmonary Langerhans cell histiocytosis: emerging concepts in pathobiology, radiology, and clinical evolution of disease. AB - Pulmonary Langerhans cell histiocytosis (PLCH) is an uncommon disorder of adult smokers associated with a significant morbidity. Arising from the aberrant accumulation of Langerhans and other immune cells, PLCH tends to cause a relatively isolated pulmonary involvement as compared to other forms of Langerhans cell (LC) and histiocytic disorders. Increased knowledge of cytokine triggers, dendritic cell trafficking, and clonality of LC populations in PLCH have resulted in an improved understanding of the pathobiology of PLCH. High resolution CT (HRCT) of the chest has led to better appreciation of nodular and cystic radiographic abnormalities characteristic of the disease. Correlation of HRCT abnormalities with lung pathologic changes has led to an improved comprehension of clinical evolution of PLCH. Current clinical predictors for PLCH outcomes remain poor, although long-term follow-up and radiologic monitoring may help to define disease progression. This review discusses advances in PLCH emphasizing the etiopathologic bases of the disease and currently available radiologic modalities for monitoring disease progression. PMID- 12740290 TI - Interpreting COPD prevalence estimates: what is the true burden of disease? AB - STUDY OBJECTIVES: To summarize the available data on COPD prevalence and assess reasons for conflicting prevalence estimates in the published literature. DESIGN: We reviewed published studies that (1) estimated COPD prevalence for a population, and (2) clearly described the methods used to obtain the estimates. RESULTS: Thirty-two sources of COPD prevalence rates, representing 17 countries and eight World Health Organization-classified regions, were identified and reviewed. Prevalence estimates were based on spirometry (11 studies), respiratory symptoms (14 studies), patient-reported disease (10 studies), or expert opinion. Reported prevalence ranged from 0.23 to 18.3%. The lowest prevalence rates (0.2 to 2.5%) were based on expert opinion. Sixteen studies had measured rates that could reasonably be extrapolated to an entire region or country. All were for Europe or North America, and most fell between 4% and 10%. CONCLUSIONS: There is considerable variation in the reported prevalence of COPD. The overall prevalence in adults appears to lie between 4% and 10% in countries where it has been rigorously measured. Some of the variation attributed to differences in risk exposure or population characteristics may be influenced by the methods and definitions used to measure disease. Spirometry is least influenced by local diagnostic practice, but it is subject to variation based on the lung function parameters selected to define COPD. PMID- 12740291 TI - Interventional pulmonary procedures: Guidelines from the American College of Chest Physicians. PMID- 12740292 TI - Non-small cell lung cancer staging techniques and endoscopic ultrasound: tissue is still the issue. AB - Non-small cell lung cancer (NSCLC) in the United States will continue to be a major public health issue, particularly as our elderly population grows. As surgery offers the best hope of cure for NSCLC, staging of NSCLC is critical because it directly impacts on the management of lung cancer. Cost, quality of life, safety, and accuracy of various staging methods all influence the clinical outcome. Staging of NSCLC is evolving due to the emergence of new and improved technologies. The objective of this article is to review the current methods used in staging of NSCLC. Currently, positron emission tomography and endoscopic ultrasound (EUS) show promise in identifying patients that may benefit from surgery. Histologic confirmation via EUS-guided fine-needle aspiration, however, may still be necessary to accurately stage the mediastinum. PMID- 12740293 TI - Severe acidosis in a patient with type 2 diabetes mellitus, hypertension, and renal failure. PMID- 12740294 TI - The genetic determinants of smoking. AB - Dependence on tobacco, like many other drug dependencies, is a complex behavior with both genetic and environmental factors contributing to the variance. The heritability estimates for smoking in twin studies have ranged from 46 to 84%, indicating a substantial genetic component to smoking. Candidate gene studies have detected functional polymorphisms in genes coding for the cytochrome P450 enzymes, and variations in these genes that lead to more rapid nicotine metabolism have been implicated in smoking. Similarly, smoking has been associated with polymorphisms in dopaminergic genes that may influence the dopamine receptor number and/or function. Animal experiments have localized specific subunits of the nicotinic receptors that may mediate the reinforcing properties of nicotine and have investigated their role in nicotine dependence. However, environmental factors have also been found to contribute to the risk of initiation and persistence of smoking. We review the scientific evidence that supports a role for genetic influences on smoking, discuss the specific genetic and neurobiological mechanisms that may mediate susceptibility to nicotine dependence, identify possible gene/environmental interactions that may be important in understanding smoking behavior, and suggest directions for future research. Insights into the genetic contributions to smoking can potentially lead to more effective strategies to reduce smoking. PMID- 12740295 TI - Respiratory findings in tobacco workers. AB - OBJECTIVES: To screen for respiratory symptoms and lung function impairment in workers occupationally exposed to tobacco dust in a tobacco-processing plant. PARTICIPANTS: One hundred twenty-one tobacco workers (97 women and 24 men) were included in the study. In addition, a group of 98 control workers (73 women and 25 men) were studied. METHODS: Acute and chronic respiratory symptoms were recorded in all tobacco workers. Lung function was measured by recording the maximum expiratory flow-volume curves on which FVC, FEV(1), and flow rates at 50% of FVC (FEF(50)) and the last 25% of FVC (FEF(25)) were read. RESULTS: There was a high prevalence of chronic respiratory symptoms among these workers, and this prevalence was significantly higher in exposed female workers than in female or male control workers (p < 0.01 and p < 0.05, respectively). Occupational asthma was recorded in 6 female tobacco workers (6.2%) and in none of the control subjects (p < 0.05). None of the male workers were found to have occupational asthma. There was also a high prevalence of acute symptoms that were noted during the work shift (particularly for nose and throat dryness, as well as eye irritation). Among these tobacco workers, the odds ratios for respiratory symptoms were frequently significant for employment and smoking among male tobacco workers but were not significant in general for female tobacco workers. The ventilatory capacity data in tobacco workers showed statistically significant reductions in FEV(1), FEF(50), and FEF(25) in relation to predicted values. These reductions were demonstrated in smokers as well as in nonsmokers. Regression analysis suggested that there were significant changes according to employment status for FVC, FEV(1), and FEF(25) in female tobacco workers. Measurements of ventilatory capacity during the course of the work shift in 38 female tobacco workers demonstrated significant mean acute reductions in FEV(1) (-5.7%), in FEF(50) (-15.7%), and in FEF(25) (-23.4%). CONCLUSIONS: Our data indicate that tobacco workers may develop respiratory disorders related to tobacco dust in their work environment. PMID- 12740296 TI - Coarctation of the aorta in an adult: problems of diagnosis and management. PMID- 12740297 TI - A new diagnostic and therapeutic approach to pericardial effusion: transbronchial needle aspiration. AB - Pericardiocentesis was introduced during the 19th century, and reached its current level of development with the introduction of two-dimensional echocardiography. Although there is general agreement that complications are rare with skilled operators, a diagnostic and therapeutic problem often occurs when there is a posterior pericardial effusion, as it is not easy to quantify by echocardiography, and difficult to drain through a percutaneous access; therefore, it is usually treated surgically. We describe a new approach to pericardial effusion by a transbronchial access through the left lower lobe bronchus (which allows both diagnosis and evacuation of abundant amounts of fluid), or through the distal trachea (for diagnostic purpose only, in the presence of pericardial effusions filling the aortic recess of the pericardium). The technique is rather easy for operators skilled in transbronchial needle aspiration, and is safe, economical, and well tolerated. PMID- 12740298 TI - Pulmonary embolism in idiopathic pulmonary fibrosis transplant recipients. AB - The objectives of the study were the assessment of the incidence of pulmonary embolism (PE) in lung transplant recipients. We performed a retrospective review of the medical records in a tertiary center lung transplant program. A total of 72 lung transplants were performed. There were seven symptomatic PE events diagnosed among six patients (group 1). All PE events were in the subgroup of patients with idiopathic pulmonary fibrosis (IPF) [6 of 23 patients (27%) vs 0 among all other patients (0%); p < 0.001]. All patients were out of the hospital, not receiving oxygen therapy, and were ambulatory at the time of the event. The median time to occurrence of the PE was 175 days posttransplant (range, 26 to 541 days). All patients who developed PEs were men. The group of IPF patients with no PEs was evenly split between genders (group 2; p < 0.009). PE patients required a longer posttransplant hospitalization (mean [+/- SD], 18.5 +/- 3.9 vs 13.5 +/- 4 days, respectively; p < 0.018). Aside from this, there was no apparent difference in patient functional status between the two groups. PEs appear to be relatively common in IPF lung transplant recipients. This should be considered in the differential diagnosis of any such patient who presents with dyspnea or hypoxia posttransplant. Patients do not appear to have been predisposed to their embolic events through lack of activity or prolonged hospital stays. PMID- 12740299 TI - Chronic eosinophilic pneumonia: a case report and national survey. AB - Few reports of chronic eosinophilic pneumonia (CEP) in the pediatric population can be found in the literature. Our patient, a 16-year-old male subject presenting with signs and symptoms of CEP, prompted a survey of pediatric pulmonary training centers in the United States to determine the prevalence of eosinophilic pneumonia. The survey showed a low prevalence of acute eosinophilic pneumonia and CEP in the pediatric population, with an overall male/female ratio of 1.6:1. PMID- 12740300 TI - A combined modality approach to recurrent cardiac sarcoma resulting in a prolonged remission: a case report. AB - We report a case of cardiac fibrosarcoma that was treated with cardiac tumor resection, chemotherapy, cardiac explantation, tumor repeat resection, and autotransplantation followed by external beam irradiation. The patient was able to achieve clinical remission and remained disease free > 2 years after the initial diagnosis. This case report demonstrates that a sustained remission from cardiac sarcoma is possible with an aggressive combined modality approach. PMID- 12740301 TI - Link between anthrax immunization and hypersensitivity pneumonitis? PMID- 12740302 TI - Chronic heart failure management: a realistic program. PMID- 12740303 TI - Breast cancer in smokers. PMID- 12740304 TI - Sitaxsentan in pulmonary arterial hypertension. PMID- 12740305 TI - Secretion management must be considered when reporting success or failure of noninvasive ventilation. PMID- 12740306 TI - Left ventricular dysfunction and sleep apnea syndrome: cause or consequence? PMID- 12740307 TI - Risk of cancers in COPD patients. PMID- 12740308 TI - Treating malignancy-related effusions. PMID- 12740311 TI - Effect of alcohol intake on muscle glycogen storage after prolonged exercise. AB - We studied the effects of alcohol intake on postexercise muscle glycogen restoration with samples from vastus lateralis being collected immediately after glycogen-depleting cycling and after a set recovery period. Six well-trained cyclists undertook a study of 8-h recovery (2 meals), and another nine cyclists undertook a separate 24-h protocol (4 meals). In each study, subjects completed three trials in crossover order: control (C) diet [meals providing carbohydrate (CHO) of 1.75 g/kg]; alcohol-displacement (A) diet (1.5 g/kg alcohol displacing CHO energy from C) and alcohol + CHO (AC) diet (C + 1.5 g/kg alcohol). Alcohol intake reduced postmeal glycemia especially in A trial and 24-h study, although insulin responses were maintained. Alcohol intake increased serum triglycerides, particularly in the 24-h study and AC trial. Glycogen storage was decreased in A diets compared with C at 8 h (24.4 +/- 7 vs. 44.6 +/- 6 mmol/kg wet wt, means +/- SE, P < 0.05) and 24 h (68 +/- 5 vs. 82 +/- 5 mmol/kg wet wt, P < 0.05). There was a trend to reduced glycogen storage with AC in 8 h (36.2 +/- 8 mmol/kg wet wt, P = 0.1) but no difference in 24 h (85 +/- 9 mmol/kg wet wt). We conclude that 1). the direct effect of alcohol on postexercise glycogen synthesis is unclear, and 2). the main effect of alcohol intake is indirect, by displacing CHO intake from optimal recovery nutrition practices. PMID- 12740312 TI - Rises in whole muscle passive tension of mammalian muscle after eccentric contractions at different lengths. AB - This is a report of experiments carried out on the medial gastrocnemius muscle of the anesthetized cat, investigating the effects of eccentric contractions carried out at different muscle lengths on the passive and active length-tension relationships. In one series of experiments, the motor supply to the muscle was divided into three approximately equal parts; in the other, whole muscles were used. Fifty eccentric contractions were carried out over different regions of the active length-tension curve for each partial or whole muscle. Active and passive length-tension curves were measured before and after the eccentric contractions. When eccentric contractions were carried out at longer lengths, there was a larger shift of the optimum length for active tension in the direction of longer muscle lengths and a larger fall in peak isometric tension. Passive tension was higher immediately after the eccentric contractions, and if the muscle was left undisturbed for 40 min, it increased further to higher values, particularly after contractions at longer lengths. A series of 20 passive stretches of the same speed and amplitude and covering the same length range as the active stretches, reduced the passive tension which redeveloped over a subsequent 40-min period. It is hypothesized that there are two factors influencing the level of passive tension in a muscle after a series of eccentric contractions. One is injury contractures in damaged muscle fibers tending to raise passive tension; the other is the presence of disrupted sarcomeres in series with still-functioning sarcomeres tending to reduce it. PMID- 12740313 TI - Effects of chronic heart failure on skeletal muscle capillary hemodynamics at rest and during contractions. AB - Chronic heart failure (CHF) reduces muscle blood flow at rest and during exercise and impairs muscle function. Using intravital microscopy techniques, we tested the hypothesis that the speed and amplitude of the capillary red blood cell (RBC) velocity (VRBC) and flux (FRBC) response to contractions would be reduced in CHF compared with control (C) spinotrapezius muscle. The proportion of capillaries supporting continuous RBC flow was less (P < 0.05) in CHF (0.66 +/- 0.04) compared with C (0.84 +/- 0.01) muscle at rest and was not significantly altered with contractions. At rest, VRBC (C, 270 +/- 62; CHF, 179 +/- 14 microm/s) and FRBC (C, 22.4 +/- 5.5 vs. CHF, 15.2 +/- 1.2 RBCs/s) were reduced (both P < 0.05) in CHF vs. C muscle. Contractions significantly (both P < 0.05) elevated VRBC (C, 428 +/- 47 vs. CHF, 222 +/- 15 microm/s) and FRBC (C, 44.3 +/- 5.5 vs. CHF, 24.0 +/- 1.2 RBCs/s) in C and CHF muscle; however, both remained significantly lower in CHF than C. The time to 50% of the final response was slowed (both P < 0.05) in CHF compared with C for both VRBC (C, 8 +/- 4; CHF, 56 +/- 11 s) and FRBC (C, 11 +/- 3; CHF, 65 +/- 11 s). Capillary hematocrit increased with contractions in C and CHF muscle but was not different (P > 0.05) between CHF and C. Thus CHF impairs diffusive and conductive O2 delivery across the rest-to-contractions transition in rat skeletal muscle, which may help explain the slowed O2 uptake on kinetics manifested in CHF patients at exercise onset. PMID- 12740314 TI - Behavior of human muscle fascicles during shortening and lengthening contractions in vivo. AB - The aim of the present study was to investigate the behavior of human muscle fascicles during dynamic contractions. Eight subjects performed maximal isometric dorsiflexion contractions at six ankle joint angles and maximal isokinetic concentric and eccentric contractions at five angular velocities. Tibialis anterior muscle architecture was measured in vivo by use of B-mode ultrasonography. During maximal isometric contraction, fascicle length was shorter and pennation angle larger compared with values at rest (P < 0.01). During isokinetic concentric contractions from 0 to 4.36 rad/s, fascicle length measured at a constant ankle joint angle increased curvilinearly from 49.5 to 69.7 mm (41%; P < 0.01), whereas pennation angle decreased curvilinearly from 14.8 to 9.8 degrees (34%; P < 0.01). During eccentric muscle actions, fascicles contracted quasi-isometrically, independent of angular velocity. The behavior of muscle fascicles during shortening contractions was believed to reflect the degree of stretch applied to the series elastic component, which decreases with increasing contraction velocity. The quasi-isometric behavior of fascicles during eccentric muscle actions suggests that the series elastic component acts as a mechanical buffer during active lengthening. PMID- 12740317 TI - Working with the private sector for child health. AB - Private sector providers are the most commonly consulted source of care for child illnesses in many countries, offering significant opportunities to expand the reach of essential child health services and products. Yet collaboration with private providers presents major challenges - the suitability and quality of the services they provide is often questionable and governments' capacity to regulate them is limited. This article assesses the actual and potential contributions of the private sector to child health, and classifies and evaluates public sector strategies to promote and rationalize the contributions of private sector actors. Governments and international organizations can use a variety of strategies to collaborate with and influence private sector actors to improve child health - including contracting, regulating, financing and social marketing, training, coordinating and informing the public. These mutually reinforcing strategies can both improve the quality of services currently delivered in the private sector, and expand and rationalize the coverage of these services. One lesson from this review is that the private sector is very heterogeneous. At the country level, feasible strategies depend on the potential of the different components of the private sector and the capacity of governments and their partners for collaboration. To date, experience with private sector strategies offers considerable promise for children's health, but also raises many questions about the feasibility and impact of these strategies. Where possible, future interventions should be designed as experiments, with careful assessment of the intervention design and the environment in which they are implemented. PMID- 12740315 TI - Prior exercise enhances passive absorption of intraduodenal glucose. AB - The purpose of this study was to assess whether a prior bout of exercise enhances passive gut glucose absorption. Mongrel dogs had sampling catheters, infusion catheters, and a portal vein flow probe implanted 17 days before an experiment. Protocols consisted of either 150 min of exercise (n = 8) or rest (n = 7) followed by basal (-30 to 0 min) and a primed (150 mg/kg) intraduodenal glucose infusion [8.0 mg x kg-1x min-1, time (t) = 0-90 min] periods. 3-O [3H]methylglucose (absorbed actively, facilitatively, and passively) and l [14C]glucose (absorbed passively) were injected into the duodenum at t = 20 and 80 min. Phloridzin, an inhibitor of the active sodium glucose cotransporter-1 (SGLT-1), was infused (0.1 mg x kg-1 x min-1) into the duodenum from t = 60-90 min with a peripheral venous isoglycemic clamp. Duodenal, arterial, and portal vein samples were taken every 10 min during the glucose infusion, as well as every minute after each tracer bolus injection. Net gut glucose output in exercised dogs increased compared with that in the sedentary group (5.34 +/- 0.47 and 4.02 +/- 0.53 mg x kg-1x min-1). Passive gut glucose absorption increased approximately 100% after exercise (0.93 +/- 0.06 and 0.45 +/- 0.07 mg x kg-1 x min-1). Transport-mediated glucose absorption increased by approximately 20%, but the change was not significant. The infusion of phloridzin eliminated the appearance of both glucose tracers in sedentary and exercised dogs, suggesting that passive transport required SGLT-1-mediated glucose uptake. This study shows 1). that prior exercise enhances passive absorption of intraduodenal glucose into the portal vein and 2). that basal and the added passive gut glucose absorption after exercise is dependent on initial transport of glucose via SGLT-1. PMID- 12740318 TI - Pro-poor health policies in poverty reduction strategies. AB - Since 1999, the International Monetary Fund and World Bank have required low income countries soliciting for debt relief and financial support to prepare a Poverty Reduction Strategy Paper (PRSP). The objective of this study is to arrive at a systematic assessment of the extent to which the first batch of interim PRSPs actually addresses the health of the poor and vulnerable. A literature study was used to design and test a semi-quantitative approach to assess the pro poor focus of health policies in national documents. The approach was applied to the existing interim proposals for 23 Highly Indebted Poor Countries. Results show that a majority of proposals lack country-specific data on the distribution and composition of the burden of disease, a clear identification of health system constraints and an assessment of the impact of health services on the population. More importantly, they make little effort to analyze these issues in relation to the poor. Furthermore, only a small group explicitly includes the interests of the poor in health policy design. Attention to policies aiming at enhancing equity in public health spending is even more limited. Few papers that include expenditure proposals also show pro-poor focused health budgets. We conclude that our systematic assessment of a new international development policy instrument, PRSP, raises strong concerns about the attributed role of health in development and the limited emphasis on the poor, the supposed primary beneficiaries of this instrument. There is a need and an opportunity for the international development community to provide assistance and inputs as poor countries shift their policy thinking from an interim stage to fully developed national policies. This paper presents a menu of analytical and policy options that can be pursued. PMID- 12740319 TI - Doubting the existence of AIDS: a barrier to voluntary HIV testing and counselling in urban Mali. AB - Qualitative research was carried out in the Malian cities of Sikasso and Bamako with a view to setting up HIV voluntary testing and counselling (VCT) services and a separate programme to enable young people to improve their sexual health. The most striking finding was that a large number of respondents said they did not believe in the existence of AIDS. Reasons for disbelief were related to the perceived lack of AIDS cases in China, the inability of the virus to be transmitted by mosquitoes and confusion about mother-to-child transmission. Highly educated individuals were very sceptical of the existence of the illness, thinking it to be a Western plot to encourage condom use in order to halt the growth of the African population. Those who were more likely to believe in the existence of the illness were less educated or uneducated people who had personally seen someone sick with AIDS, often when they had been on labour migration to Cote d'Ivoire where HIV prevalence is higher. Respondents thought it likely that this scepticism will limit the use of VCT services. Other reasons for the potential non-use of services included the fact that some people lacked confidence in the competence of the laboratory technicians and were afraid that those testing positive would be highly stigmatized by the community. Thus, widespread awareness-raising campaigns are needed before any centres can be set up. Participatory education programmes are required to address HIV in the context of other health risks. This would allow people to inter-actively shape the debate about HIV/AIDS to fit their own needs. Currently, they are presented with information about the illness in a unidirectional manner via the media or health educators which seems to fuel their scepticism. PMID- 12740320 TI - Infant-feeding practices of mothers of known HIV status in Lusaka, Zambia. AB - BACKGROUND: Between 25 and 44% of mother-to-child transmission (MTCT) of the human immunodeficiency virus (HIV) occurs through breastfeeding. As a result, feeding guidelines for infants of HIV-infected mothers are being formulated in many resource-poor countries. The impact of introducing these guidelines on mothers' actual feeding practices has not previously been examined. Infant feeding practices of mothers of known HIV status who should have received advice during pre- and post-test HIV counselling were assessed and compared with those of uninfected mothers. METHODS: Mothers of infants aged 2-12 months, 55 HIV infected and 85 HIV-uninfected, were recruited from the HIV Family Support Unit in Lusaka, Zambia. HIV status was known to 121 of these mothers, who had all received pre- and post-test HIV counselling. Feeding practices were determined by verbal questionnaire. RESULTS: All mothers breastfed but only 35% of infants below 4 months were exclusively breastfed (received breast milk only). HIV infected mothers introduced fluids and weaned their infants significantly earlier than HIV-uninfected mothers (p = 0.03 and p = 0.002, respectively). Infants of HIV-infected mothers had significantly lower weight for age Z (WAZ) scores indicating poorer nutritional or health status (p = 0.004). Commercial formula milk and cow's milk were used by 36 mothers as breast milk substitutes, and were introduced at a median age of 2.5 months. Thirteen mothers gave cow's milk, and no mother added water to cow's milk (as recommended), with two adding sugar and four adding salt. CONCLUSION: Infant-feeding practices of HIV-infected mothers differed significantly from HIV-uninfected mothers, and this may contribute to their poorer growth. Paradoxically these mothers feeding practice could be putting these infants at greater risk of both non-HIV-related morbidity and HIV transmission, as early introduction of foods other than breast milk may increase MTCT. PMID- 12740322 TI - The impact of the 1997-98 East Asian economic crisis on health and health care in Indonesia. AB - This article identifies the effects of the 1997-98 East Asian economic crisis on health care use and health status in Indonesia. The article places the findings in the context of a framework showing the complex cause and effect relationships underlying the effects of economic downturns on health and health care. The results are based on primary analysis of Indonesian household survey data and review of a wide range of sources from the Indonesian government and international organizations. Comparisons are drawn with the effects of the crisis in Thailand. The devaluation of the Indonesian currency, the Rupiah, led to inflation and reduced real public expenditures on health. Households' expenditures on health also decreased, both in absolute terms and as a percentage of overall spending. Self-reported morbidity increased sharply from 1997 to 1998 in both rural and urban areas of Indonesia. The crisis led to a substantial reduction in health service utilization during the same time period, as the proportion of household survey respondents reporting an illness or injury that sought care from a modern health care provider declined by 25%. In contrast to Indonesia, health care utilization in Thailand actually increased during the crisis, corresponding to expansion in health insurance coverage. The results suggest that social protection programmes play a critical role in protecting populations against the adverse effects of economic downturns on health and health care. PMID- 12740321 TI - Targeted subsidy for malaria control with treated nets using a discount voucher system in Tanzania. AB - During the last decade insecticide-treated nets have become a key strategy for malaria control. Social marketing is an appealing tool for getting such nets to poor rural African communities who are most afflicted by malaria. This approach usually involves subsidized prices to make nets and insecticide more affordable and help establish a commercial market. We evaluated a voucher system for targeted subsidy of treated nets in young children and pregnant women in two rural districts of southern Tanzania. Qualitative work involved focus group discussions with community leaders, male and female parents of children under 5 years. In-depth interviews were held with maternal and child health clinic staff and retail agents. Quantitative data were collected through interviewing more than 750 mothers of children under 5 years during a cluster sample survey of child health. The voucher return rate was extremely high at 97% (7720/8000). However, 2 years after the start of the scheme awareness among target groups was only 43% (45/104), and only 12% of women (12/103; 95% CI 4-48%) had used a voucher towards the cost of a net. We found some evidence of increased voucher use among least poor households, compared with the poorest households. On the basis of these results we renewed our information, education and communication (IEC) campaign about vouchers. Discount vouchers are a feasible system for targeted subsidies, although a substantial amount of time and effort may be needed to achieve high awareness and uptake - by which we mean the proportion of eligible women who used the vouchers - among those targeted. Within a poor society, vouchers may not necessarily increase health equity unless they cover a high proportion of the total cost: since some cash is needed when using a voucher as part-payment, poorer women among the target group are likely to have lower uptake than richer women. The vouchers have two important additional functions: strengthening the role of public health services in the context of a social marketing programme and forming an IEC tool to demonstrate the group at most risk of severe malaria. PMID- 12740323 TI - Is the Colombian health system reform improving the performance of public hospitals in Bogota? AB - Many countries are experimenting with public hospital reform - both increasing the managerial autonomy with which hospitals conduct their affairs, and separating 'purchaser' and 'provider' sides of the health system, thus increasing the degree of market pressure brought to bear on hospitals. Evidence suggesting that such reform will improve hospital performance is weak. From a theoretical perspective, it is not clear why public hospitals should be expected to behave like firms and seek to maximize profits as this model requires. Empirically, there is very slight evidence that such reforms may improve efficiency, and reason to be concerned about their equity implications. In Colombia, an ambitious reform programme includes among its measures the attempt to universalize a segmented health system, the creation of a purchaser-provider split and the transformation of public hospitals into 'autonomous state entities'. By design, the Colombian reform programme avoids the forces that produce equity losses in other developing countries. This paper reports the results of a study that has tried to track hospital performance in other dimensions in the post-reform period in Bogota. Trends in hospital inputs, production and productivity, quality and patient satisfaction are presented, and qualitative data based on interviews with hospital workers are analyzed. The evidence we have been able to collect is capable of providing only a partial response to the study question. There is some evidence of increased activity and productivity and sustained quality despite declining staffing levels. Qualitative data suggest that hospital workers have noticed considerable changes, which include greater responsiveness to patients but also a heavier administrative burden. It is difficult to attribute specific causality to all of the changes measured and this reflects the inherent difficulty of judging the effects of large-scale reform programmes as well as weaknesses and gaps in the data available. PMID- 12740325 TI - Public participation in health planning and priority setting at the district level in Uganda. AB - OBJECTIVE: To explore the experiences of the public and leaders with participatory planning and priority setting in health, in a decentralized district in Uganda. METHODOLOGY: An exploratory qualitative approach, involving in-depth interviews with health planners at the national, district and community levels (n = 12), and five group discussions at community level with women (two groups), men, youths and adolescents (n = 51). The analysis adapted some principles from grounded theory. The five levels of the participation framework by Rifkin (1991) were used to assess the actual level of participation in the study population. RESULTS: Uganda has established structures for participatory planning. Within this context, district level respondents reported to have gained decision-making powers, but were concerned about the degree of financial independence they had. The national level respondents were concerned about the capacity of the districts to absorb their new roles. Actual involvement of the public in priority setting and poor communication between the different levels of the decentralization system, despite the existing structures, were additional concerns. Public participation is mainly through representatives. Majority participation is mainly at health benefits and programme activity levels. Decision-making, monitoring and evaluation, and implementation are still dominated by the locally elected leaders due to reported economic, social and cultural barriers that hinder the participation of the rest of the public. PMID- 12740324 TI - Contracting for health services: an evaluation of recent reforms in Nicaragua. AB - Nicaragua has embarked on a reform of the way in which publicly provided medical care is organized and financed. A principal feature of the reforms includes a decentralization of decision-making authority coupled with an increase in local accountability. Local decision-making authority has been increased by allowing managers more freedom to allocate inputs. Accountability has been strengthened by stipulating what is expected of hospitals and health centres in the form of performance agreements, and tying rewards (i.e. bonuses) to the satisfaction of these requirements provides incentives. This paper provides a critical assessment of these reforms, and presents some early evidence of their effects. PMID- 12740326 TI - Implementing a new health management information system in Uganda. AB - This paper reports on research investigating the health management information system (HMIS) implementation process in Uganda, utilizing the diffusion of innovation and dynamic equilibrium organizational change models. Neither perspective guided the HMIS development process. Instead, technological issues, rather than wider organizational issues, dominated the planned change. The need to consider the organizational context when changing information systems arises because the process is more complex than some practitioners have realized, when attempting to understand the causes of information management problems and developing HMIS in low-income countries. In particular, information system developers had not acknowledged that they were promoting an informational approach to management when they promoted a change from a centralized reporting system to a HMIS supporting use of information at the level of collection. Strategies to facilitate this approach were not advocated. Organizational theory can contribute to the diffusion of innovation framework. It has yielded an integration of Rogers's diffusion of innovation framework and Leavitt's concept of organizational forces in equilibrium. The diffusion framework describes the process, but the organizational model has given the context and reason for aspects of the process. The diffusion model does not predict what needs to change within the organization when a particular innovation is introduced, or how much. The addition of the organizational model has helped. These frameworks can facilitate the introduction of future information management innovations and allow practitioners to perceive their introduction as a staged process needing to be managed. Implications for practice are identified. PMID- 12740327 TI - Postnatal anaemia: neglected problems and missed opportunities in Uganda. AB - Anaemia is a neglected disorder that affects a huge proportion of women, particularly in developing countries. In a cross-sectional study among 349 mothers attending child clinics in Tororo District, eastern Uganda, who had delivered within 12 months prior to the study, the magnitude and prevalence of anaemia, important risk factors for anaemia and the usefulness of clinical examination and patient history as a screening tool were determined. Approximately two-thirds (64.4%) of the women were anaemic, with 55 (15.8%) suffering from moderate to severe anaemia (Hb < 10.0 g/dl) and 169 (48.6%) with mild anaemia (10.0-11.9 g/dl). Five (1.4%) women suffered from severe anaemia (Hb < 7.0 g/dl). Only iron supplementation and excessive bleeding during or after delivery emerged as risk factors using multinominal regression modelling. Lack of iron supplementation was a factor for mild anaemia (odds ratio (OR) 2.6; 95% confidence interval (CI) 1.5-4.2), but not for moderate to severe anaemia. Excessive bleeding was a risk factor for moderate to severe anaemia (OR 2.3; 95% CI 1.1-4.7), but not for mild anaemia. The sensitivity of using clinical signs to detect anaemia (hb < 12.0 g/dl) was relatively high (0.74; 95% CI 0.7-0.8), but with a low specificity (0.4; 95% CI 0.3-0.6). Although anaemia in postnatal women is widespread, the health care system had missed the opportunities to effectively address it, such as through the implementation of the WHO policy recommendation for iron and folic acid supplementation, improvement of obstetric services and, despite its imperfection, screening for anaemia using clinical signs. PMID- 12740328 TI - Rates of public investment for road safety in developing countries: case studies of Uganda and Pakistan. AB - OBJECTIVE: This paper assesses the magnitude of public investment in road safety in Uganda and Pakistan. METHODS: The study reviewed government budgetary records on expenditure for road safety for each country, as well as World Bank estimates of bilateral and NGO assistance directed to road safety. The authors interviewed key informants in each government who would know about public or NGO activity on road safety. RESULTS: Budgetary expenditure on road safety at all levels of government in Uganda and Pakistan is $0.09 and $0.07 per capita respectively. DISCUSSION: The scale of public activity in road safety in Uganda and Pakistan is extremely limited. If there are diminishing returns to scale for road safety investments, this would suggest that the potential effectiveness of properly chosen safety measures could never be higher. CONCLUSION: Large reductions in morbidity and mortality are likely if investment in road safety is expanded. PMID- 12740330 TI - Direct measurement of acid permeation into rat oesophagus. AB - BACKGROUND AND AIMS: The early responses of the oesophageal mucosa to acid perfusion may predict subsequent pathology. Mucosal responses to luminal acid may result either from acid permeating through the mucosa or from other unknown transduction mechanisms. In order to better understand the dynamics of acid permeation into the oesophageal mucosa, we measured interstitial pH (pH(int)) of the oesophageal basal epithelial layer, pre-epithelial layer thickness, and blood flow in rats in vivo during luminal acid challenge. A novel confocal microscopic technique was used in vitro to measure pH(int) from defined cellular sites in response to luminal and basolateral acidification. METHODS: 5-(and-6) Carboxyfluorescein (CF) and carboxy-seminapthorhodofluor-1 (SNARF-1) fluorescence was used to measure pH(int) by conventional and confocal microscopy, respectively, in urethane anaesthetised rats. Pre-epithelial layer thickness was measured optically with carbon particles as markers. Blood flow was measured with laser Doppler flowmetry. RESULTS: Luminal acidification failed to alter pH(int) in vivo and in vitro, but pH(int) was lowered by modest serosal acidification. Pre-epithelial layer thickness and blood flow increased significantly during luminal surface acid perfusion. Indomethacin had no effect on any acid related response. CONCLUSION: In this first dynamic measurement of oesophageal acid permeation and pre-epithelial layer thickness, pH(int) was preserved in spite of high luminal acidity by two complementary techniques. Despite the apparent permeability barrier to acid permeation, oesophageal blood flow and thickness responded to luminal acid perfusion. PMID- 12740331 TI - Relevance of ineffective oesophageal motility during oesophageal acid clearance. AB - BACKGROUND: Oesophageal clearance of acid reflux consists of an initial volume clearance followed by neutralisation of the acidified mucosa by swallowed saliva (chemical clearance). Ineffective oesophageal motility (IOM), a frequent finding in patients with gastro-oesophageal reflux disease (GORD), has been claimed to underlie prolonged acid clearance by affecting oesophageal emptying and saliva transport. Intraluminal impedance allows non-radiological monitoring of movement of oesophageal liquids. AIMS: To evaluate the relevance of IOM during oesophageal volume and chemical clearance using combined pH impedance measurements. SUBJECTS: Impedance was validated with fluoroscopy to study volume clearance in three healthy subjects. Acid clearance tests were performed in 10 healthy subjects in the upright and supine positions, before and after oesophageal peristaltic disruption with sildenafil 50 mg. METHODS: After instillation of an acid bolus, simultaneous manometry, pH, and impedance were used to study oesophageal motility, chemical clearance, and volume clearance, respectively. RESULTS: Impedance allowed assessment of volume clearance accurately, showing a strong correlation with fluoroscopy (r(2)=0.89). Sildenafil provoked a graded impairment in oesophageal motility in healthy subjects without affecting saliva secretion. In the upright position, volume clearance was slightly prolonged only with severe IOM (>80% abnormal peristaltic sequences). In the supine position, severe IOM significantly prolonged chemical and volume clearance. Moderate IOM (30-80% abnormal peristalsis) had no effect. With normal peristalsis and moderate IOM, clearance times were similar in the upright and supine positions. Severe IOM however had a greater impact on clearance in the supine than in the upright position. CONCLUSION: Ineffective oesophageal motility has little effect on oesophageal clearance during upright acid reflux. With supine reflux, only severe IOM is associated with prolonged oesophageal clearance. PMID- 12740332 TI - On the existence and location of cardiac mucosa: an autopsy study in embryos, fetuses, and infants. AB - BACKGROUND: The incidence of gastric cardiac adenocarcinoma has increased in the last decades. Gaining insight into the pathogenesis of this lesion is hampered by the limited knowledge of the origin and histology of cardiac mucosa (CM). Currently, the location, extent, and even the existence of CM are controversial. AIMS: We studied the development of the gastro-oesophageal junction (GOJ) in embryos, fetuses, and infants to clarify if CM is a normal structure at birth and where it is located. SUBJECTS: Twenty one autopsy cases were evaluated ranging in age from 13 weeks' gestational age (GA) to seven months. METHODS: The distal oesophagus and proximal part of the stomach were embedded entirely. Serial sections were stained with haematoxylin-eosin and alcian blue/periodic acid Schiff. The following parameters were measured: length of abdominal oesophagus; length of columnar lined oesophagus; length of CM; and distance from CM to angle of His. RESULTS: CM was present in all evaluated sections. Its mean length varied throughout gestation. A maximum value was reached at a GA of 16 weeks (1.2 mm). After term delivery it was very short (0.3-0.6 mm). CM was proximal to, or straddled, the angle of His in all cases. During gestation, the mucin staining pattern of the CM was to a high degree similar to that of the developing pyloric mucosa. CONCLUSIONS: CM develops during pregnancy and is present at birth as a normal structure. If the angle of His is taken as a landmark for the GOJ, CM is located in the distal oesophagus. PMID- 12740333 TI - Helicobacter pylori infection induces duodenitis and superficial duodenal ulcer in Mongolian gerbils. AB - BACKGROUND: There is no direct evidence for an animal model of Helicobacter pylori induced duodenal ulcer. AIM: In this study we evaluated the roles of bacterial strain and age of experimental animals in induction of duodenitis and duodenal ulcer in Mongolian gerbils after H pylori infection. METHODS: Specific pathogen free Mongolian gerbils were inoculated orally with three bacterial strains (H pylori ATCC 43504, TN2GF4, and K-6, a clinical isolate from a patient with gastric cancer in our clinic). These strains have both the cagA gene and VacA. Five week old gerbils were used to emulate prematurity infection and 14 week old animals were used as mature test subjects. Animals were observed for 12 weeks after inoculation. Interleukin 8 (IL-8) production in gastric epithelial cells (MKN74) after coculture with the H pylori strains was measured by ELISA. RESULTS: Gastritis and gastric ulcers were found in all gerbils infected with the three strains. However, duodenitis and gastric metaplasia were seen more frequently in gerbils infected with TN2GF4 and K-6 strains than in the ATCC 43504 infected or control groups (p<0.05). Superficial duodenal ulcers with severe duodenitis and gastric metaplasia were found in two gerbils inoculated at 14 weeks with the TN2GF4 strain but none at five weeks. The TN2GF4 strain stimulated significantly higher levels of IL-8 than ATCC 43504 and K6 strains (p=0.0039). CONCLUSIONS: When injected into adult Mongolian gerbils, a specific strain (TN2GF4) of H pylori can induce duodenitis with gastric metaplasia and superficial duodenal ulcers. Induction of duodenal ulcer in an animal model fulfills the requirements of Koch's postulates for establishing a role for H pylori as a causative agent. PMID- 12740334 TI - Evaluation of a novel monoclonal enzyme immunoassay for detection of Helicobacter pylori antigen in stool from children. AB - BACKGROUND: Reliable non-invasive methods for detection of Helicobacter pylori infection are required to investigate the incidence, transmission, and clearance of infection in childhood. AIM: To evaluate a new monoclonal enzyme immunoassay (EIA) (FemtoLab H pylori Cnx) for detection of H pylori antigen in stool in a large cohort of children compared with invasive diagnostic methods and the (13)C urea breath test. PATIENTS AND METHODS: A total of 302 symptomatic previously untreated children (aged 0.5-18.7 years; 148 girls) were recruited at three centres. H pylori status was defined by results of culture, histology, the rapid urease test, and the (13)C urea breath test. Stool samples were investigated locally by the EIA using two different production lots. According to the manufacturer's recommendations, an optical density (OD) of 0.150 was used as a cut off value. RESULTS: OD values clearly differentiated between the 92 H pylori infected and the 210 non-infected children (median (5th-95th percentiles) 2.729 (0.232->4.000) v 0.021 (0.009-0.075)). Only two false positive and two false negative results occurred, giving a sensitivity, specificity, positive predictive value, and negative predictive value of 98%, 99%, 98%, and 99%, respectively. No significant relation was found between age and OD values in infected or non infected children. CONCLUSIONS: The monoclonal stool antigen EIA was excellent in diagnosing H pylori infection in symptomatic children. Accuracy was independent of the laboratory, production lot used, or the child's age. Because only 18/116 children <6 years of age were infected with H pylori, further validation of the test is needed in young infected children. PMID- 12740336 TI - Gastrin activates nuclear factor kappaB (NFkappaB) through a protein kinase C dependent pathway involving NFkappaB inducing kinase, inhibitor kappaB (IkappaB) kinase, and tumour necrosis factor receptor associated factor 6 (TRAF6) in MKN-28 cells transfected with gastrin receptor. AB - BACKGROUND: We previously reported that gastrin induces expression of CXC chemokines through activation of nuclear factor kappaB (NFkappaB) in gastric epithelial cells that express gastrin receptor. AIMS: To clarify gastrin receptor mediated signals leading to activation of NFkappaB. METHODS: MKGR26 cells were created by transfecting gastrin receptor cDNA into MKN-28 cells. Degradation of inhibitor kappaB (IkappaB) and phosphorylation of protein kinase C (PKC)-delta were both detected by western blot analysis. NFkappaB activation was determined by luciferase assay and electrophoretic mobility shift analysis. RESULTS: Gastrin induced degradation of IkappaB-alpha and activation of NFkappaB, which was abolished by the selective gastrin receptor antagonist L-740,093 and the general PKC inhibitor GF109203X. Gastrin induced phosphorylation of PKC-delta, and its inhibitor rottlerin partially suppressed NFkappaB activation. However, the mitogen activated protein kinase (MAPK) kinase inhibitor PD98059, p38 MAPK inhibitor SB203580, and tyrphostin AG1478 had no effect on NFkappaB activation. Introduction of the dominant negative mutant of IkappaB kinase, of NFkappaB inducing kinase, and of tumour necrosis factor receptor associated factor 6 (TRAF6), but not that of TRAF2, inhibited gastrin induced activation of NFkappaB. CONCLUSIONS: Gastrin activates NFkappaB via a PKC dependent pathway which involves IkappaB kinase, NFkappaB inducing kinase, and TRAF6. PMID- 12740335 TI - Gastric intestinal metaplasia as detected by a monoclonal antibody is highly associated with gastric adenocarcinoma. AB - BACKGROUND: Some forms of gastric intestinal metaplasia (GIM) may be precancerous but the cellular phenotype that predisposes to gastric carcinogenesis is not well characterised. Mucin staining, as a means of differentiating GIM, is difficult. A monoclonal antibody, mAb Das-1 (initially called 7E(12)H(12)), whose staining is phenotypically specific to colon epithelium, was used to investigate this issue. METHODS: Using mAb Das-1, by a sensitive immunoperoxidase assay, we examined histologically confirmed GIM specimens from two countries, the USA and Japan. A total of 150 patients comprised three groups: group A, GIM (fields away from the cancer area) from patients with gastric carcinoma (n=60); group B, GIM with chronic gastritis (without gastric carcinoma) (n=72); and group C, chronic gastritis without GIM (n=18). RESULTS: Fifty six of 60 (93%) patients with GIM (both goblet and non-goblet metaplastic cells) from group A reacted intensely with mAb Das-1. Cancer areas from the same 56 patients also reacted. In contrast, 25/72 (35%) samples of GIM from patients in group B reacted with mAb Das-1 (group A v B, p<0.0001). None of the samples from group C reacted with the mAb. CONCLUSIONS: Reactivity of mAb Das-1 is clinically useful to simplify and differentiate the phenotypes of GIM. The colonic phenotype of GIM, as identified by mAb Das-1, is strongly associated with gastric carcinoma. PMID- 12740337 TI - Incidence of gastroduodenal ulcers in patients with rheumatoid arthritis after 12 weeks of rofecoxib, naproxen, or placebo: a multicentre, randomised, double blind study. AB - BACKGROUND: Previous studies in patients with osteoarthritis have suggested that the selective cyclooxygenase (COX)-2 inhibitor rofecoxib results in less gastrointestinal damage than non-selective non-steroidal antiinflammatory drugs (NSAIDs). This study compared the incidence of endoscopically detected gastroduodenal ulcers in rheumatoid arthritis patients treated with rofecoxib or a non-selective NSAID. METHODS: In this multicentre, randomised, double blind, 12 week study, patients with rheumatoid arthritis were allocated to rofecoxib 50 mg once daily (n=219), naproxen 500 mg twice daily (n=220), or placebo (n=221). Endoscopy was performed at baseline and at six and 12 weeks. Lifetable analysis and log rank tests were used to analyse the incidence of gastroduodenal ulcers > or =3 mm. Gastric or duodenal ulcers > or =5 mm and erosions were also evaluated as secondary end points. Tolerability was assessed by adverse events. RESULTS: The cumulative incidence of ulcers > or =3 mm at 12 weeks was significantly higher in patients on naproxen (25.5%) than in patients receiving rofecoxib (6.8%; difference 18.7% (95% confidence interval (CI) 11.7%, 25.7%); p<0.001) or placebo (2.9%; difference 22.6% (95% CI 16.1%, 29.1%); p<0.001). The difference between rofecoxib (6.8%) and placebo (2.9%) did not reach statistical significance (p=0.066). Results were similar for ulcers > or =5 mm and for mean changes from baseline in the number of gastroduodenal erosions. The overall incidence of clinical adverse events was similar among treatment groups (61% of patients on placebo, 62% in patients on rofecoxib, and 66% in patients on naproxen). CONCLUSIONS: Rofecoxib 50 mg daily (twice the dose recommended for this patient population) resulted in a lower incidence of endoscopically detected gastroduodenal ulcers and erosions than treatment with naproxen 500 mg twice daily. PMID- 12740338 TI - Extracellular MUC3 mucin secretion follows adherence of Lactobacillus strains to intestinal epithelial cells in vitro. AB - BACKGROUND: Mucins are large complex glycoproteins that protect intestinal mucosal surfaces by limiting access of environmental matter to their epithelial cells. Several mucin genes have been described, including MUC3 that is a membrane associated mucin of the small intestine. Increased MUC3 mRNA transcription is induced by incubation of intestinal epithelial cells with a Lactobacillus strain known to be adherent to them. AIMS: To determine whether increased epithelial cell MUC3 mucin expression in response to Lactobacillus strains results in increased extracellular secretion of MUC3 mucins and the importance of epithelial cell adherence in modulation of MUC3 mucin expression. METHODS: HT29 cells grown to enhance expression of MUC3 mucins were incubated with selected Lactobacillus strains. Spent cell culture medium was collected for detection of secreted MUC3 mucins using dot blot immunoassay with a generated MUC3 antibody. Post-incubation HT29 cell RNA was collected for analysis of MUC3 expression by northern blot analysis using a MUC3 cDNA probe. In vitro binding studies using Lactobacillus strains incubated alone or coincubated with enteropathogenic Escherichia coli strain E2348/69 were used for adherence and inhibition of adherence studies, respectively. RESULTS: Lactobacillus strains with minimal ability to adhere to HT29 cells failed to induce upregulation of mucin gene expression. There was a direct correlation between upregulation of MUC3 mucin mRNA expression and extracellular secretion of MUC3 mucin. The same Lactobacillus strains that increased extracellular secretion of MUC3 mucin led to reduced adherence of enteropathogen E coli E2348/69 during coincubation experiments. CONCLUSION: Probiotic microbes induce MUC3 mucin transcription and translation with extracellular secretion of the MUC3 mucins. Epithelial cell adherence enhances the effects of probiotics on eukaryotic mucin expression. PMID- 12740339 TI - Mobilisation of enterocyte fat stores by oral glucose in humans. AB - BACKGROUND AND AIMS: When a high fat oral load is followed several hours later by further ingestion of nutrients, there is an early postprandial peak in plasma triacylglycerol (TG). The aim of this study was to investigate the location and release of lipid from within the gastrointestinal tract. METHODS: Ten healthy patients undergoing oesopho-gastro-duodenoscopy (OGD) were recruited. At t=0, all patients consumed a 50 g fat emulsion and at t=5 hours they consumed either water or a 38 g glucose solution. OGD was performed at t=6 hours and jejunal biopsy samples were evaluated for fat storage. A subgroup of five subjects then underwent a parallel metabolic study in which postprandial lipid and hormone measurements were taken during an identical two meal protocol. RESULTS: Following oral fat at t=0, samples from patients that had subsequently ingested glucose exhibited significantly less staining for lipid within the mucosa and submucosa of the jejunum than was evident in patients that had consumed only water (p=0.028). There was also less lipid storage within the cytoplasm of enterocytes (p=0.005) following oral glucose. During the metabolic study, oral glucose consumed five hours after oral fat resulted in a postprandial peak in plasma TG, chylomicron-TG, and apolipoprotein B48 concentration compared with oral water. CONCLUSION: After a fat load, fat is retained within the jejunal tissue and released into plasma following glucose ingestion, resulting in a peak in chylomicron-TG which has been implicated in the pathogenesis of atherosclerosis. PMID- 12740340 TI - Card15 gene overexpression in mononuclear and epithelial cells of the inflamed Crohn's disease colon. AB - BACKGROUND: Crohn's disease is one of the principal human chronic inflammatory bowel diseases. Although its aetiology is still unknown, its complex pathogenesis has environmental, immunological, and genetic determinants. CARD15 is the first susceptibility gene implicated in the predisposition to Crohn's disease and is known to be expressed only in monocytes. However, its expression in situ has not yet been studied. AIMS: To analyse the tissue distribution of CARD15 and identify cells producing CARD15 in samples of colon from patients with Crohn's disease and control subjects. PATIENTS AND METHODS: We analysed CARD15 gene expression in surgical specimens of colon from eight children with Crohn's disease and nine controls by immunohistochemistry, in situ hybridisation, and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: We showed that CARD15 was present only in the cytoplasm of macrophages in the normal colon. Increased CARD15 expression was detected in Crohn's disease lesions. There were more CARD15 positive cells in Crohn's disease lesions than in uninvolved areas. Both intestinal epithelial cells, macrophages, and their derivatives overproduced CARD15 in Crohn's disease. To further assess CARD15 expression by intestinal epithelial cells, we performed RT-PCR on freshly isolated intestinal epithelial cells, and showed that these cells isolated from Crohn's disease samples contained more CARD15 mRNA than intestinal epithelial cells from controls. CONCLUSIONS: We have demonstrated that colonic involvement in active Crohn's disease is associated with increased CARD15 gene expression in both macrophages and intestinal epithelial cells. Therefore, this deregulation can affect the host environment interaction and thus contribute to the pathogenesis of this disease. PMID- 12740341 TI - Neutrophil derived human S100A12 (EN-RAGE) is strongly expressed during chronic active inflammatory bowel disease. AB - BACKGROUND: Intestinal inflammation in Crohn's disease (CD) and ulcerative colitis (UC) is characterised by an influx of neutrophils into the intestinal mucosa. S100A12 is a calcium binding protein with proinflammatory properties. It is secreted by activated neutrophils and interacts with the multiligand receptor for advanced glycation end products (RAGE). Promising anti-inflammatory effects of blocking agents for RAGE have been reported in murine models of colitis. AIMS: To investigate expression and serum concentrations of S100A12 in inflammatory bowel disease (IBD). METHODS: We performed immunohistochemical studies and immunofluorescence microscopy in biopsies from patients with CD and UC. S100A12 serum concentrations were analysed using a sandwich ELISA. RESULTS: Immunohistochemical studies revealed profound expression of S100A12 in inflamed intestinal tissue from IBD patients whereas no expression was found in tissue from healthy controls. Staining for S100A12 during chronic active CD and UC was restricted to infiltrating neutrophils. Serum S100A12 levels were significantly elevated in patients with active CD (470 (125) ng/ml; p<0.001, n=30) as well as those with active UC (400 (120) ng/ml; p<0.01, n=15) compared with healthy controls (75 (15) ng/ml; n=30). Even in inactive disease, elevated serum concentrations were found, at least in CD. S100A12 levels were well correlated with disease activity in CD and UC. CONCLUSIONS: We demonstrated that neutrophil derived S100A12 is strongly upregulated during chronic active IBD, suggesting an important role during the pathogenesis of IBD. Serum S100A12 may serve as a useful marker for disease activity in patients with IBD. PMID- 12740342 TI - Non-genomic regulation of intermediate conductance potassium channels by aldosterone in human colonic crypt cells. AB - BACKGROUND: Aldosterone has a rapid, non-genomic, inhibitory effect on macroscopic basolateral K(+) conductance in the human colon, reducing its capacity for Cl(-) secretion. The molecular identity of the K(+) channels constituting this aldosterone inhibitable K(+) conductance is unclear. AIM: To characterise the K(+) channel inhibited by aldosterone present in the basolateral membrane of human colonic crypt cells. METHODS: Crypts were isolated from biopsies of healthy sigmoid colon obtained during colonoscopy. The effect of aldosterone on basolateral K(+) channels, and the possible involvement of Na(+):H(+) exchange, were studied by patch clamp techniques. Total RNA from isolated crypts was subjected to reverse transcriptase-polymerase chain reaction (RT-PCR) using primers specific to intermediate conductance K(+) channels (KCNN4) previously identified in other human tissues. RESULTS: In cell attached patches, 1 nmol/l aldosterone significantly decreased the activity of intermediate conductance (27 pS) K(+) channels by 31%, 53%, and 54% after 1, 5 and 10, minutes, respectively. Increasing aldosterone concentration to 10 nmol/l produced a further 56% decrease in channel activity after five minutes. Aldosterone 1-10 nmol/l had no effect on channel activity in the presence of 20 micro mol/l ethylisopropylamiloride, an inhibitor of Na(+):H(+) exchange. RT-PCR identified KCNN4 mRNA, which is likely to encode the 27 pS K(+) channel inhibited by aldosterone. CONCLUSION: Intermediate conductance K(+) channels (KCNN4) present in the basolateral membranes of human colonic crypt cells are a target for the non-genomic inhibitory effect of aldosterone, which involves stimulation of Na(+):H(+) exchange, thereby reducing the capacity of the colon for Cl(-) secretion. PMID- 12740344 TI - Influence of dietary factors on colorectal cancer survival. AB - BACKGROUND: Diet has been identified as a major determinant of colorectal cancer (CRC) but little is known of its influence on CRC survival. AIMS: To study the influence of dietary factors on survival in patients who had undergone potentially curative CRC surgery. PATIENTS: Among 171 patients included in a case control study of CRC aetiological factors, 10 year survival data on 148 patients who underwent resection of the tumour for potential cure were obtained from a Registry of Digestive Tumours. METHODS: Tertiles of food and nutrient intakes were entered into Cox proportional hazards survival models, controlling for age, sex, tumour stage, and tumour location. RESULTS: Only five year survival was influenced by the pre-diagnosis diet. High energy intake, as a result of high carbohydrate, protein, and lipid intake, was strongly related to increased survival. Five year relative risk of death for the highest versus the two lowest tertiles of energy intake was 0.18 (95% confidence interval 0.07; 0.44). This effect was similar in both sexes, for the colon and for the rectum. It was stronger in patients with N+/M+ tumours (relative risk 0.06) than in those with less advanced tumours (relative risk 0.37; stage-energy interaction term non significant). No specific food or nutrient could be identified as having prognostic significance. CONCLUSIONS: Whether high energy intake selects less severe tumoral clones or modifies antitumoral immunity remains unclear. Larger series need to be investigated before conducting intervention studies but our findings should prompt nutritional follow up in CRC patients. PMID- 12740343 TI - Relations between amount and type of alcohol and colon and rectal cancer in a Danish population based cohort study. AB - BACKGROUND: There may be a weak association between total alcohol intake and colorectal cancer but the effect of different types of alcohol and effect on colon subsites have not been investigated satisfactorily. AIMS: To investigate the relationship between amount and type of alcohol and the risk of colon and rectal cancer. SUBJECTS: A population based cohort study with baseline assessment of weekly intake of beer, wine, and spirits, smoking habits, body mass index, educational level, and leisure time physical activity in Copenhagen, Denmark. The study included a random sample of 15 491 men and 13 641 women, aged 23-95 years. Incident cases of colorectal cancer were identified in the nationwide Danish Cancer Register. RESULTS: During a mean follow up of 14.7 years, we observed 411 colon cancers and 202 rectal cancers. We observed a dose-response relationship between alcohol and rectal cancer. Drinkers of more than 41 drinks a week had a relative risk of rectal cancer of 2.2 (95% confidence limits 1.0-4.6) compared with non-drinkers. Drinkers of more than 14 drinks of beer and spirits a week, but not wine, had a risk of 3.5 (1.8-6.9) of rectal cancer compared with non drinkers, while those who drank the same amount of alcohol but including more than 30% of wine had a risk of 1.8 (1.0-3.2) of rectal cancer. No relation between alcohol and colon cancer was found when investigating the effects of total alcohol, beer, wine, and spirits, and percentage of wine of total alcohol intake. CONCLUSION: Alcohol intake is associated with a significantly increased risk of rectal cancer but the risk seems to be reduced when wine is included in the alcohol intake. PMID- 12740346 TI - Child-Pugh versus MELD score in predicting survival in patients undergoing transjugular intrahepatic portosystemic shunt. AB - BACKGROUND: In patients undergoing transjugular intrahepatic portosystemic shunt (TIPS), prognostic scores may identify those with a poor prognosis or even those with a clear survival benefit. The Child-Pugh score (CPS) is well established but several drawbacks have led to development of the model of end stage liver disease (MELD). AIM: The aim of the study was to compare the predictive power of CPS and MELD, to validate the original MELD formula, and to assess the predictive value of the determinants used in the two prognostic scores outside of a study setting. PATIENTS: A total of 501 patients underwent elective TIPS placement and 475 patients fulfilled the inclusion criteria. METHODS: Data of all patients undergoing elective TIPS in one university hospital and four community hospitals in Vienna, Austria, between 1991 and 2001, were analysed retrospectively. The main statistical tests were Cox proportional hazards regression model, the log rank test, Kaplan-Meier analysis, and concordance c statistics. RESULTS: Median follow up was 5.2 years and median survival was 4.6 years. During follow up, 230 patients died, 75 within three months after TIPS placement. In stepwise proportional hazards analyses, independent predictors of death were creatinine level, bilirubin level, age, and refractory ascites. MELD was better in predicting survival in a stepwise Cox model but both scores were equally predictive in c statistics for one month, three month, and one year survival. Renal function was the strongest independent predictor of survival. CONCLUSIONS: Although MELD was the primary predictor of overall survival in multivariate analysis, c statistics showed that both scores can be used for patients undergoing TIPS with equal accuracy. For assessing prognosis in patients undergoing TIPS implantation, there seems little reason to replace the well established Child-Pugh score. PMID- 12740345 TI - Administration of protegrin peptide IB-367 to prevent endotoxin induced mortality in bile duct ligated rats. AB - BACKGROUND: Postoperative morbidity in patients with obstructive jaundice remains high because of increased susceptibility to endotoxin and the inflammatory cascade. AIMS: An experimental study was designed to investigate the efficacy of protegrin peptide IB-367, an antimicrobial positively charged peptide, in neutralising Escherichia coli 0111:B4 lipopolysaccharide (LPS) in bile duct ligated rats. METHODS: Adult male Wistar rats were injected intraperitoneally with 2 mg/kg E coli 0111:B4 LPS one week after sham operation or bile duct ligation (BDL). Six groups were studied: sham with placebo, sham with 120 mg/kg tazobactam-piperacillin (TZP), sham with 1 mg/kg IB-367, BDL with placebo, BDL with 120 mg/kg TZP, and BDL with 1 mg/kg IB-367. RESULTS: Main outcome measures were: endotoxin and tumour necrosis factor alpha (TNF-alpha) concentrations in plasma, evidence of bacterial translocation in blood and peritoneum, and lethality. After LPS, TNF-alpha plasma levels were significantly higher in BDL rats compared with sham operated animals. IB-367 caused a significant reduction in plasma endotoxin and TNF-alpha concentrations compared with placebo and TZP treated groups. In contrast, both TZP and IB-367 significantly reduced bacterial growth compared with saline treatment. Finally, LPS induced 60% and 55% lethality in BDL placebo and TZP treated rats and no lethality in sham operated rats, while only IB-367 significantly reduced lethality to 10%. CONCLUSIONS: By virtue of its dual antimicrobial and antiendotoxin properties, IB-367 could be an interesting compound to inhibit bacterial translocation and endotoxin release in obstructive jaundice. PMID- 12740347 TI - Detection of submucosal gastric fundal varices with multi-detector row CT angiography. AB - BACKGROUND AND AIM: The diagnosis of submucosal fundal varices is challenging. Currently, endoscopy and endoscopic ultrasound (EUS) are considered most useful for this purpose. The aim of this study was to evaluate if multi-detector row CT (MDCT) angiography contributes to the diagnosis of submucosal fundal varices. PATIENTS AND METHODS: Twenty two patients with endoscopically suspected fundal varices were prospectively included in the study. All patients underwent EUS and MDCT angiography. Levels of agreement between EUS and MDCT angiography for the detection of submucosal and perigastric fundal varices were evaluated by three blinded independent readers. In addition, variceal size and location, as well as afferent and efferent vessels of the submucosal varices, were determined. RESULTS: Good or excellent image quality of MDCT angiography was obtained in 21/22 patients (95%). Based on EUS, submucosal varices were detected in 16 of 22 patients (73%) and perigastric varices in 22/22 patients (100%). Using MDCT angiography, the presence of submucosal varices was confirmed in all of these 16 patients by all three readers. Perigastric varices were also confirmed in all 22 patients by all three readers. In addition, all three readers noted the presence of a submucosal varix in an additional patient which was not detected on initial EUS. MDCT angiography showed an excellent interobserver reliability with regard to variceal diameter (kappa=0.90) and variceal location (kappa=0.94). Based on MDCT angiography, afferent and efferent vessels of submucosal varices included the left gastric vein in 11 (65%), the posterior/short gastric veins in 15 (88%), gastrorenal shunts in 10 (59%), the left inferior phrenic vein in six (35%), and the left pericardiophrenic vein in six (35%) of 17 patients. CONCLUSIONS: MDCT angiography is equivalent to EUS in terms of detection and characterisation of fundal varices, in particular with regard to the distinction between submucosal and perigastric fundal varices. PMID- 12740349 TI - Rapidly progressive adenomatous polyposis in a patient with germline mutations in both the APC and MLH1 genes: the worst of two worlds. AB - The two most common inherited forms of colorectal cancer are familial adenomatous polyposis and hereditary non-polyposis colorectal cancer. Simultaneous inheritance of both an APC gene mutation and a mismatch repair gene (for example, MLH1) mutation has never been described. In the present case report, we report rapidly progressive adenomatous polyposis in a 10 year old boy with a germline frame shift mutation in the APC gene and a germline splice site mutation in the MLH1 gene. Immunohistochemical investigations showed abnormal expression of beta catenin in early adenomas with low grade dysplasia, attributed to the APC gene mutation. Subsequent loss of function of the MLH1 gene, as shown by absent immunostaining of its protein in adenomas with high grade dysplasia, may well have caused the rapid progression to high grade dysplasia in many of the adenomas. PMID- 12740348 TI - A 10 year follow up study of patients transplanted for autoimmune hepatitis: histological recurrence precedes clinical and biochemical recurrence. AB - BACKGROUND AND AIM: Autoimmune hepatitis (AIH) has been reported to recur after orthotopic liver transplantation (OLT) in 10-35% of patients in small series with a short follow up. The aim of the present study was to examine the clinical and histological outcome more than 10 years after OLT for AIH. PATIENTS AND METHODS: Seventeen women with a mean age of 30 (12) years at the time of OLT, selected from among 44 patients transplanted for AIH, were followed for more than 10 years. The criteria for definite AIH, as established by the International Autoimmune Hepatitis Group, were met in every case. Liver biopsies were performed 1, 2, 5, and 10 years after OLT, and when indicated by abnormal liver function tests. Specimens were examined for evidence of recurrent AIH, namely interface hepatitis, lobular activity, portal lymphoplasmocytic infiltration, and fibrosis. Other signs of recurrence included hypertransaminasaemia, serum autoantibodies, and the response to steroid reintroduction or significant steroid dose increments. RESULTS: AIH recurred in 7 (41%) of 17 patients. In four patients histological abnormalities were detected by means of protocol biopsies 1-5 years before the onset of biochemical abnormalities. Two patients developed severe recurrences after 10 and 15 years, respectively, and required treatment with steroids and tacrolimus. In the other three patients histological recurrence was detected 0.6-3 years post-OLT, concomitantly with biochemical abnormalities. CONCLUSIONS: AIH recurred in 41% of patients followed for more than 10 years after OLT. As histological signs preceded biochemical abnormalities in four patients (23.5%), regular liver biopsy is warranted after OLT. Detection of isolated histological signs may call for closer follow up and/or a change in immunosuppressive therapy. PMID- 12740350 TI - Second infection with a different hepatitis C virus genotype in a intravenous drug user during interferon therapy. AB - The prevalence of antibodies against hepatitis C virus (anti-HCV) among intravenous drug users (IVDU) has consistently been very high. Cross challenge studies in chimpanzees provide evidence that reinfection with different HCV strains may occur. In humans, reinfection with different HCV strains has been reported in multitransfused haemophiliacs and recently in IVDU but no case has been reported while on interferon (IFN) therapy. We report on a 22 year old woman who was treated with IFN alpha for HCV genotype 3a chronic infection. At six months, HCV RNA was undetectable by reverse transcription-polymerase chain reaction. In October 1997, while still on IFN, she developed an acute hepatitis after an intravenous drug injection and HCV genotype 1a infection was identified using genotyping and sequencing methods. IFN therapy was continued until August 1998, and in January 1999 HCV-RNA was not detectable. Our case indicates that the previous HCV infection might have prevented development of chronicity. An alternative explanation is that IFN, while not preventing acute hepatitis C, may prevent chronicity. The risk of multiple infection in IVDU underlines the need for preventive strategies. PMID- 12740352 TI - Hepatoma in a young adult: to biopsy or not to biopsy? PMID- 12740351 TI - Attenuated familial adenomatous polyposis presenting as ampullary adenocarcinoma. AB - The risk of periampullary cancer in patients with classic familial adenomatous polyposis (FAP) is significantly increased compared with the general population. However, the incidence of this extracolonic manifestation in attenuated FAP (AFAP) is unknown. We report the case of a 38 year old woman with no known family history of polyposis or colorectal cancer, who presented with ampullary adenocarcinoma. Diagnosis of AFAP was made only after evaluation of the patient's extended family history and genetic testing. This case report suggests that AFAP should be included in the differential diagnosis of patients with ampullary/duodenal tumours. PMID- 12740354 TI - Paris staging system for primary gastrointestinal lymphomas. PMID- 12740353 TI - Chronic pancreatitis: the perspective of pain generation by neuroimmune interaction. AB - Chronic pancreatitis (CP) is an inflammatory, often painful, disease of the exocrine pancreas which leads to exocrine insufficiency. The pathophysiology of pain in CP is incompletely understood. Several hypotheses have been advanced, including pancreatic and extrapancreatic causes. Here, the different pain hypotheses are discussed and evidence is presented that neuroimmune interactions are significant in the pathogenesis of pain generation and inflammation in CP. A better understanding of the complex cellular and molecular mechanisms of neuroimmune interactions should offer possibilities for innovative therapy and long term disease prevention. PMID- 12740355 TI - A new cause for CA19.9 elevation: heavy tea consumption. PMID- 12740356 TI - Expression of thiopurine methyltransferase in South Asians. PMID- 12740357 TI - Non-alcoholic steatohepatitis and hepatic steatosis in patients with adult onset growth hormone deficiency. PMID- 12740358 TI - Mutant K-ras2 in serum. PMID- 12740359 TI - Fatigue is associated with high circulating leptin levels in chronic hepatitis C. PMID- 12740360 TI - The C-terminally truncated NuoL subunit (ND5 homologue) of the Na+-dependent complex I from Escherichia coli transports Na+. AB - The NADH:quinone oxidoreductase (complex I) from Escherichia coli acts as a primary Na+ pump. Expression of a C-terminally truncated version of the hydrophobic NuoL subunit (ND5 homologue) from E. coli complex I resulted in Na+ dependent growth inhibition of the E. coli host cells. Membrane vesicles containing the truncated NuoL subunit (NuoLN) exhibited 2-4-fold higher Na+ uptake activity than control vesicles without NuoLN. Respiratory proton transport into inverted vesicles containing NuoLN decreased upon addition of Na+, but was not affected by K+, indicating a Na+-dependent increase of proton permeability of membranes in the presence of NuoLN. The His-tagged NuoLN protein was solubilized, enriched by affinity chromatography, and reconstituted into proteoliposomes. Reconstituted His6-NuoLN facilitated the uptake of Na+ into the proteoliposomes along a concentration gradient. This Na+ uptake was prevented by EIPA (5-(N-ethyl N-isopropyl)-amiloride), which acts as inhibitor against Na+/H+ antiporters. PMID- 12740361 TI - Characterization of a heparan sulfate 3-O-sulfotransferase-5, an enzyme synthesizing a tetrasulfated disaccharide. AB - Heparan sulfate d-glucosaminyl 3-O-sulfotransferases (3-OSTs) catalyze the transfer of sulfate from 3'-phosphoadenosine 5'-phosphosulfate (PAPS) to position 3 of the glucosamine residue of heparan sulfate and heparin. A sixth member of the human 3-OST family, named 3-OST-5, was recently reported (Xia, G., Chen, J., Tiwari, V., Ju, W., Li, J.-P., Malmstrom, A., Shukla, D., and Liu, J. (2002) J. Biol. Chem. 277, 37912-37919). In the present study, we cloned putative catalytic domain of the human 3-OST-5 and expressed it in insect cells as a soluble enzyme. Recombinant 3-OST-5 only exhibited sulfotransferase activity toward heparan sulfate and heparin. When incubated heparan sulfate with [35S]PAPS, the highest incorporation of35S was observed, and digestion of the product with a mixture of heparin lyases yielded two major35S-labeled disaccharides, which were determined as DeltaHexA-GlcN(NS,3S,6S) and DeltaHexA(2S)-GlcN(NS,3S) by further digestion with 2-sulfatase and degradation with mercuric acetate. However, when used heparin as acceptor, we identified a highly sulfated disaccharide unit as a major product. This had a structure of DeltaHexA(2S)-GlcN(NS,3S,6S). Quantitative real time PCR analysis revealed that 3-OST-5 was highly expressed in fetal brain, followed by adult brain and spinal cord, and at very low or undetectable levels in the other tissues. Finally, we detected a tetrasulfated disaccharide unit in bovine intestinal heparan sulfate. To our knowledge, this is the first report to describe not only the natural occurrence of tetrasulfated disaccharide unit but also the enzymatic formation of this novel structure. PMID- 12740362 TI - Heat shock protein 27 controls apoptosis by regulating Akt activation. AB - Activation of the serine-threonine kinase Akt by cytokines, chemokines, and bacterial products delays constitutive neutrophil apoptosis, resulting in a prolonged inflammatory response. We showed previously that Akt exists in a signaling complex with p38 MAPK, MAPK-activated protein kinase-2 (MAPKAPK-2), and heat shock protein-27 (Hsp27); and Hsp27 dissociates from the complex upon neutrophil activation. To better understand the regulation of this signaling module, the hypothesis that Akt phosphorylation of Hsp27 regulates its interaction with Akt was tested. The present study shows that Akt phosphorylated Hsp27 on Ser-82 in vitro and in intact cells, and phosphorylation of Hsp27 resulted in its dissociation from Akt. Additionally, the interaction between Hsp27 and Akt was necessary for activation of Akt in intact neutrophils. Constitutive neutrophil apoptosis was accelerated by sequestration of Hsp27 from Akt, and this enhanced rate of apoptosis was reversed by introduction of constitutively active recombinant Akt. Our results define a new mechanism by which Hsp27 regulates apoptosis, through control of Akt activity. PMID- 12740363 TI - Identification of a novel voltage-driven organic anion transporter present at apical membrane of renal proximal tubule. AB - A novel transport protein with the properties of voltage-driven organic anion transport was isolated from pig kidney cortex by expression cloning in Xenopus laevis oocytes. A cDNA library was constructed from size-fractionated poly(A)+ RNA and screened for p-aminohippurate (PAH) transport in high potassium medium. A 1856-base pair cDNA encoding a 467-amino acid peptide designated as OATV1 (voltage-driven organic anion transporter 1) was isolated. The predicted amino acid sequence of OATV1 exhibited 60-65% identity to those of human, rat, rabbit, and mouse sodium-dependent phosphate cotransporter type 1 (NPT1), although OATV1 did not transport phosphate. The homology of this transporter to known members of the organic anion transporter family (OAT family) was about 25-30%. OATV1 mediated PAH transport was affected by the changes in membrane potential. The transport was Na+-independent and enhanced at high concentrations of extracellular potassium and low concentrations of extracellular chloride. Under the voltage clamp condition, extracellularly applied PAH induced outward currents in oocytes expressing OATV1. The current showed steep voltage dependence, consistent with the voltage-driven transport of PAH by OATV1. The PAH transport was inhibited by various organic anions but not by organic cations, indicating the multispecific nature of OATV1 for anionic compounds. This transport protein is localized at the apical membrane of renal proximal tubule, consistent with the proposed localization of a voltage-driven organic anion transporter. Therefore, it is proposed that OATV1 plays an important role to excrete drugs, xenobiotics, and their metabolites driven by membrane voltage through the apical membrane of the tubular epithelial cells into the urine. PMID- 12740364 TI - Roles for sphingolipid biosynthesis in mediation of specific programs of the heat stress response determined through gene expression profiling. AB - Previous studies have demonstrated roles for de novo production of sphingolipids in Saccharomyces cerevisiae in the regulation of the transient cell cycle arrest and nutrient permease degradation associated with the heat stress response, suggesting multiple functions for yeast sphingolipids in this response. We, therefore, sought to determine the generalized involvement of sphingolipids in the heat stress response by using microarray hybridization of RNA isolated from heat-stressed cultures of the mutant strain lcb1-100, which is unable to produce sphingolipids in response to heat. Approximately 70 genes showed differential regulation during the first 15 min of heat stress in the lcb1-100 strain compared with the wild type strain, indicating a requirement for de novo sphingolipid biosynthesis for proper regulation of these genes during heat stress. Grouping these genes into functional categories revealed several pathways, including some in which sphingolipids were previously suspected to play a role, such as stress response pathways and cell cycle regulation. Hierarchical clustering analysis revealed sphingolipid involvement in regulation of tRNA synthesis and metabolic genes and transporters. Additionally, the microarray results demonstrated novel sphingolipid involvement in transcriptional regulation of pathways of translation and cell wall organization and biogenesis. Our results demonstrate a broad reaching effect of sphingolipids in the yeast heat stress response and suggest that the mechanism of sphingolipid involvement in several cellular pathways occurs via sphingolipid-mediated regulation of message levels. PMID- 12740365 TI - ARF tumor suppressor induces mitochondria-dependent apoptosis by modulation of mitochondrial Bcl-2 family proteins. AB - A tumor suppressor gene product, ARF, sensitizes cells to apoptosis in the presence of appropriate collateral signals. In this study, we analyzed the mechanism of ARF-dependent apoptosis and demonstrated that ARF induces mitochondria-dependent apoptosis in p53 wild-type, ARF/p16-null cells. We also found that ARF evokes cytochrome c release from mitochondria, decreases mitochondrial membrane potential, and activates pro-caspase-9 to induce apoptosis. Our findings suggest that this apoptotic cellular modulation is brought about by up-regulation of the proapoptotic Bcl-2 family proteins Bax and Bim and down-regulation of antiapoptotic Bcl-2 in mitochondrial fractions. Additionally, ARF seems to down-regulate Bcl-2 in a p53-dependent manner while up regulating Bax/Bim via a p53-independent pathway. PMID- 12740366 TI - The digestive food vacuole of the malaria parasite is a dynamic intracellular Ca2+ store. AB - The acidic food vacuole of Plasmodium falciparum has been the subject of intense scientific investigation in the 40 years since its role in the digestion of host hemoglobin was first suggested. This proposed role has important implications for the complex host-parasite inter-relationship and also for the mode of action of several of the most effective antimalarial drugs. In addition, adaptive changes in the physiology of this organelle are implicated in drug resistance. Here we show that in addition to these functions, the digestive food vacuole of the malaria parasite is a dynamic internal store for free Ca2+, a role hitherto unsuspected. With the aid of live-cell laser scanning confocal imaging, spatiotemporal studies revealed that maintenance of elevated free Ca2+ in the digestive food vacuole (relative to cytosolic levels) is achieved by a thapsigargin (and cyclopiazonic acid)-sensitive Ca2+-pump in cooperation with a H+-dependent Ca2+ transporter. Redistribution of free cytosolic and vacuolar Ca2+ during parasite growth also suggests that vacuolar Ca2+ plays an essential role in parasite morphogenesis. These data imply that the digestive food vacuole of the malaria parasite is functionally akin to the vacuole of plants (tonoplast) and the small electron-dense granules of some parasites (acidocalcisomes) whereby H+-coupled Ca2+ transport is involved in ion transport, Ca2+ homeostasis, and signal transduction. These findings have significant implications for parasite development, antimalarial drug action, and mechanisms of drug resistance. PMID- 12740368 TI - Structure-based analysis of high pressure adaptation of alpha-actin. AB - Deep-sea fishes occur to depths of several thousand meters, and at these abyssal depths encounter pressures that shallower living fishes cannot tolerate. Tolerance of abyssal pressures by deep-sea fish is likely to depend in part on adaptive modifications of proteins. However, the types of structural modifications to proteins that allow function at high pressure have not been discovered. To elucidate the mechanisms of protein adaptation to high pressure, we cloned the alpha-skeletal actin cDNAs from two abyssal Coryphaenoides species, C. armatus and C. yaquinae, and identified three amino acid substitutions, V54A or L67P, Q137K, and A155S, that distinguish these abyssal actins from orthologs of alpha-actin from non-abyssal Coryphaenoides. These substitutions, Q137K and A155S, prevent the dissociation reactions of ATP and Ca2+ from being influenced by high pressure. In particular, the lysine residue at position 137 results in a much smaller apparent volume change in the Ca2+ dissociation reaction. The V54A or L67P substitution reduces the volume change associated with actin polymerization and has a role in maintaining the DNase I activity of actin at high pressure. Together, these results indicate that a few amino acid substitutions in key functional positions can adaptively alter the pressure sensitivity of a protein. PMID- 12740367 TI - Contrasting membrane interaction mechanisms of AP180 N-terminal homology (ANTH) and epsin N-terminal homology (ENTH) domains. AB - Epsin and AP180/CALM are endocytotic accessory proteins that have been implicated in the formation of clathrin-coated pits. Both proteins have phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2)-binding domains in their N termini, but these domains are structurally and functionally different. To understand the basis of their distinct properties, we measured the PtdIns(4,5)P2-dependent membrane binding of the epsin N-terminal homology (ENTH) domain and the AP180 N-terminal homology (ANTH) domain by means of surface plasmon resonance and monolayer penetration techniques and also calculated the effect of PtdIns(4,5)P2 on the electrostatic potential of these domains. PtdIns(4,5)P2 enhances the electrostatic membrane association of both domains; however, PtdIns(4,5)P2 binding exerts distinct effects on their membrane dissociation. Specifically, PtdIns(4,5)P2 induces the membrane penetration of the N-terminal alpha-helix of the ENTH domain, which slows the membrane dissociation of the domain and triggers the membrane deformation. These results provide the biophysical explanation for the membrane bending activity of epsin and its ENTH domain. PMID- 12740369 TI - Mechanistic understanding of an altered fidelity simian immunodeficiency virus reverse transcriptase mutation, V148I, identified in a pig-tailed macaque. AB - We have recently reported that the reverse transcriptase (RT) of SIVMNE 170 (170), which is a representative viral clone of the late symptomatic phase of infection with the parental strain, SIVMNE CL8 (CL8), has a largely increased fidelity, compared with the CL8 RT. In the present study, we analyzed the mechanistic alterations of the high fidelity 170 RT variant. First, we found that among several 170 RT mutations, only one, V148I, is solely responsible for the fidelity increase over the CL8 RT. This V148I mutation lies near the Gln-151 residue that we recently found is important to the low fidelity of RT and the binding of incoming dNTPs. Second, we compared dNTP binding affinity (Kd) and catalysis (kpol) of the CL8 RT and the CL8-V148I RT using pre-steady state kinetic analysis. In this experiment, the high fidelity CL8-V148I RT has largely decreased binding to both correct and incorrect dNTP without altering kpol. The fidelity increase imparted by the V148I mutation is likely because of the major reduction seen in RT binding to dNTPs. This parallels our findings with the Q151N mutant. Third, site-directed mutagenesis targeting amino acid residue 148 has revealed that a valine amino acid at this position is essential to RT infidelity. Based on these findings, we discuss possible structural impacts of residue 148 (and mutations at this site) on the interaction of RT with incoming dNTPs and infer how alterations in these properties may relate to viral replication and fitness. PMID- 12740370 TI - Ets-1 protects vascular smooth muscle cells from undergoing apoptosis by activating p21WAF1/Cip1: ETS-1 regulates basal and and inducible p21WAF1/Cip: ETS 1 regulates basal and inducible p21WAF1/Cip1 transcription via distinct cis acting elements in the p21WAF/Cip1 promoter. AB - The cyclin-dependent kinase inhibitor (CKI) p21WAF1/Cip1 is regulated at the level of transcription by nuclear factors such as the co-activator p300. It is presently unknown whether the Ets family of transcription factors control p21WAF1/Cip1 gene expression. Ets-1 inhibits apoptosis in vascular smooth muscle cells as determined by both fluorescein isothiocyanate-linked annexin V/propidium iodide staining of cells and fluorescence-activated cell sorting analysis and quantitative cytoplasmic histone-associated internucleosomal DNA fragmentation. p21WAF1/Cip1 can play a mitogenic and anti-apoptotic role in smooth muscle cells. Using transient transfection and Western blot analysis, we determined that Ets-1 activates p21WAF1/Cip1 transcription and protein expression. Electrophoretic mobility shift assays revealed that Ets-1 interacts selectively with the 1350GGAA-1347 Ets element in the p21WAF1/Cip1 promoter. Mutation of this element reduced basal and Ets1-inducible p21WAF1/Cip1 promoter-dependent expression. In contrast, the -1577GGAT-1574 motif mediates basal but not Ets-1 activation of the p21WAF1/Cip1 promoter. Co-immunoprecipitation and co-transfection analysis showed that Ets-1 binds p300 and cooperatively activates p21WAF1/Cip1 transcription. The phenotypic importance of Ets-1 regulation of p21WAF1/Cip1 was demonstrated by the capacity of antisense p21WAF1/Cip1 strategies to block Ets-1-inhibition of apoptosis and inhibit Ets-1-induction of proliferation. PMID- 12740371 TI - AMP-activated protein kinase regulates HNF4alpha transcriptional activity by inhibiting dimer formation and decreasing protein stability. AB - AMP-activated protein kinase (AMPK) is the central component of a cellular signaling system that regulates multiple metabolic enzymes and pathways in response to reduced intracellular energy levels. The transcription factor hepatic nuclear factor 4alpha (HNF4alpha) is an orphan nuclear receptor that regulates the expression of genes involved in energy metabolism in the liver, intestine, and endocrine pancreas. Inheritance of a single null allele of HNF4alpha causes diabetes in humans. Here we demonstrate that AMPK directly phosphorylates HNF4alpha and represses its transcriptional activity. AMPK-mediated phosphorylation of HNF4alpha on serine 304 had a 2-fold effect, reducing the ability of the transcription factor to form homodimers and bind DNA and increasing its degradation rate in vivo. These results demonstrate that HNF4alpha is a downstream target of AMPK and raise the possibility that one of the effects of AMPK activation is reduced expression of HNF4alpha target genes. PMID- 12740372 TI - Importin alpha nuclear localization signal binding sites for STAT1, STAT2, and influenza A virus nucleoprotein. AB - Proteins actively transported into the nucleus via the classical nuclear import pathway contain nuclear localization signals (NLSs), which are recognized by the family of importin alpha molecules. Importin alpha contains 10 armadillo (arm) repeats, of which the N-terminal arm repeats 2-4 have been considered as the "major" NLS binding site. Interferon-activated, dimerized signal transducers and activators of transcription (STAT1 and STAT2) directly bind to importin alpha5 via a dimeric nonclassical NLS. Here we show by site-directed mutagenesis that the very C-terminal arm repeats 8 and 9 of importin alpha5 form a unique binding site for STAT1 homodimers and STAT1-STAT2 heterodimers. Influenza A virus nucleoprotein also contains a nonclassical NLS that is recognized by the C terminal NLS binding site of importin alpha5, comprising arm repeats 7-9. Binding of influenza A virus nucleoprotein to importin alpha3 also occurs via the C terminal arm repeats. Simian virus 40 large T antigen instead binds to the major N-terminal arm repeats of importin alpha3, indicating that one importin alpha molecule is able to use either its N- or C-terminal arm repeats for binding various NLS containing proteins. PMID- 12740373 TI - Modulation of mitogenic activity of fibroblast growth factors by inorganic polyphosphate. AB - The proliferation of normal human fibroblast cells was enhanced by the addition of inorganic polyphosphate (poly(P)) into culture media. The mitogenic activities of acidic fibroblast growth factor (FGF-1) and basic fibroblast growth factor (FGF-2) were also enhanced by poly(P). A physical interaction between poly(P) and FGF-2 was observed, and FGF-2 was both physically and functionally stabilized by poly(P). Furthermore, poly(P) facilitated the FGF-2 binding to its cell surface receptors. Because poly(P) is widely distributed in mammalian tissues, it may be a spontaneous modulator of FGFs. PMID- 12740374 TI - Molecular interaction and enzymatic activity of macrophage migration inhibitory factor with immunorelevant peptides. AB - Disulfide reduction is an important step in antigen processing for HLA class II restricted T cell responses. Migration inhibitory factor (MIF) is a member of the thioredoxin family and has been classically defined as a cytokine. Using enzyme linked immunosorbent assay and CD analysis, here we describe the binding to MIF of two peptides, hepatitis B surface antigen (HBsAg) and insulin B (InsB) with high affinity for HLA class II allo-types, HLA-DP2 and HLA-DQ8, respectively. At neutral pH, cysteinylated InsB was a substrate for MIF thiol reductase activity, as assessed by mass spectroscopy/electrospray analysis. Finally, a biologically active form of MIF co-immunopurified with mature forms of HLA DP2/15, and a peptide derived from the HLA-DP beta1 helix could be used for affinity purification of MIF. The possibility that MIF participates in class II antigen presentation and/or as a chaperone is discussed. PMID- 12740375 TI - Arabidopsis histone acetyltransferase AtGCN5 regulates the floral meristem activity through the WUSCHEL/AGAMOUS pathway. AB - Histone acetyltransferases, which are able to acetylate histone and non-histone proteins, play important roles in gene regulation. Many histone acetyltransferases are related to yeast Gcn5, a component of two transcription regulatory complexes SAGA and ADA. In this work, by characterizing a mutation in the Arabidopsis GCN5 gene (AtGCN5) we studied the regulatory function of this gene in controlling floral meristem activity. We show that in addition to pleiotropic effects on plant development, this mutation also leads to the production of terminal flowers. The flowers show homeotic transformations of petals into stamens and sepals into filamentous structures and produce ectopic carpels. The phenotypes correlate to an expansion of the expression domains within floral meristems of the key regulatory genes WUSCHEL (WUS) and AGAMOUS (AG). These results suggest that AtGCN5 is required to regulate the floral meristem activity through the WUS/AG pathway. This study brings new elements on the elucidation of specific developmental pathways regulated by AtGCN5 and on the control mechanism of meristem regulatory gene expression. PMID- 12740376 TI - The dynamic dimerization of the yeast ADP/ATP carrier in the inner mitochondrial membrane is affected by conserved cysteine residues. AB - The ADP/ATP carrier (AAC) that facilitates the translocation of ATP made in mitochondria is inserted at the inner mitochondrial membrane by the TIM10-TIM22 protein import system. Here we addressed the state of the AAC precursor during insertion (stage IV of import) and identified residues of the carrier important for dimerization. By a combination of (i) import of a mix of His-tagged and untagged versions of AAC either 35S-labeled or unlabeled, (ii) import of a tandem covalent dimer AAC into wild-type mitochondria, and (iii) import of monomeric AAC into mitochondria expressing only the tandem covalent dimer AAC, we found that the stage IV intermediate is a monomer, and this stage is probably the rate limiting step of insertion in the membrane. Subsequent dimerization occurs extremely rapidly (within less than a minute). The incoming monomer dimerizes with monomeric endogenous AAC suggesting that the AAC dimer is very dynamic. Conserved Cys residues were found not to affect insertion significantly, but they are crucial for the dimerization process to obtain a functional carrier. PMID- 12740377 TI - Macrophage endothelial nitric-oxide synthase autoregulates cellular activation and pro-inflammatory protein expression. AB - Expression of inducible nitric-oxide (NO) synthase (iNOS) and "high-output" production of NO by macrophages mediates many cytotoxic actions of these immune cells. However, macrophages have also been shown to express a constitutive NOS isoform, the function of which remains obscure. Herein, bone marrow-derived macrophages (BMDMOs) from wild-type and endothelial NOS (eNOS) knock-out (KO) mice have been used to assess the role of this constitutive NOS isoform in the regulation of macrophage activation. BMDMOs from eNOS KO animals exhibited reduced nuclear factor-kappaB activity, iNOS expression, and NO production after exposure to lipopolysaccharide (LPS) as compared with cells derived from wild type mice. Soluble guanylate cyclase (sGC) was identified in BMDMOs at a mRNA and protein level, and activation of cells with LPS resulted in accumulation of cyclic GMP. Moreover, the novel non-NO-based sGC activator, BAY 41-2272, enhanced BMDMO activation in response to LPS, and the sGC inhibitor 1H (1,2,4)oxadiazolo(4,3-a)quinoxalin-1-one attenuated activation. These observations provide the first demonstration of a pathophysiological role for macrophage eNOS in regulating cellular activation and suggest that NO derived from this constitutive NOS isoform, in part via activation of sGC, is likely to play a pivotal role in the initiation of an inflammatory response. PMID- 12740378 TI - Co-stimulation of mGluR5 and N-methyl-D-aspartate receptors is required for potentiation of excitatory synaptic transmission in hippocampal neurons. AB - In the central nervous system, excitatory synaptic transmission is mediated by the neurotransmitter glutamate and its receptors. Interestingly, stimulation of group I metabotropic glutamate receptors (mGluRs) can either enhance or depress synaptic transmission at CA1 hippocampal synapses. Here we report that co activation of mGluR5, a member of the group I mGluR family, and N-methyl-d aspartate receptors (NMDARs) potentiates NMDAR currents and induces a long lasting enhancement of excitatory synaptic transmission in primary cultured hippocampal neurons. Unexpectedly, activation of mGluR5 alone fails to enhance evoked NMDAR currents and synaptic alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate receptor (AMPAR) AMPAR currents. The observed potentiation requires an mGluR5-induced, inositol 1,4,5-trisphosphate receptor-mediated mobilization of intracellular Ca2+, which acts in concert with a protein kinase C, calcium-activated tyrosine kinase cascade to induce a long lasting enhancement of NMDAR and AMPAR currents. PMID- 12740379 TI - PASK (proline-alanine-rich STE20-related kinase), a regulatory kinase of the Na-K Cl cotransporter (NKCC1). AB - Although the phosphorylation-dependent activation of the Na-K-Cl cotransporter (NKCC1) has been previously well documented, the identity of the kinase(s) responsible for this regulation has proven elusive. Recently, Piechotta et al. (Piechotta, K., Lu, J., and Delpire, E. (2002) J. Biol. Chem. 277, 50812-50819) reported the binding of PASK (also referred as SPAK (STE20/SPS1-related proline alanine-rich kinase)) and OSR1 (oxidative stress response kinase) to cation chloride cotransporters KCC3, NKCC1, and NKCC2. In this report, we show that overexpression of a kinase inactive, dominant negative (DN) PASK mutant drastically reduces both shark (60 +/- 5%) and human (80 +/- 3%) NKCC1 activation. Overexpression of wild type PASK causes a small (sNKCC1 22 +/- 8% p < 0.05, hNKCC1 12 +/- 3% p < 0.01) but significant increase in shark and human cotransporter activity in HEK cells. Importantly, DNPASK also inhibits the phosphorylation of two threonines, contained in the previously described N terminal regulatory domain. We additionally show the near complete restoration of NKCC1 activity in the presence of the protein phosphatase type 1 inhibitor calyculin A, demonstrating that DNPASK inhibition results from an alteration in kinase/phosphatase dynamics rather than from a decrease in functional cotransporter expression. Coimmunoprecipitation assays confirm PASK binding to NKCC1 in transfected HEK cells and further suggest that this binding is not a regulated event; neither PASK nor NKCC1 activity affects the association. In cells preloaded with 32Pi, the phosphorylation of PASK, but not DNPASK, coincides with that of NKCC1 and increases 5.5 +/- 0.36-fold in low [Cl]e. These data conclusively link PASK with the phosphorylation and activation of NKCC1. PMID- 12740380 TI - Biosynthesis of dTDP-3-acetamido-3,6-dideoxy-alpha-D-galactose in Aneurinibacillus thermoaerophilus L420-91T. AB - The glycan chain of the S-layer protein of Aneurinibacillus thermoaerophilus L420 91T (DSM 10154) consists of d-rhamnose and 3-acetamido-3,6-dideoxy-d-galactose (d Fucp3NAc). Thymidine diphosphate-activated d-Fucp3NAc serves as precursor for the assembly of structural polysaccharides in Gram-positive and Gram-negative organisms. The biosynthesis of dTDP-3-acetamido-3,6-dideoxy-alpha-d-galactose (dTDP-d-Fucp3NAc) involves five enzymes. The first two steps of the reaction are catalyzed by enzymes that are part of the well studied dTDP-l-rhamnose biosynthetic pathway, namely d-glucose-1-phosphate thymidyltransferase (RmlA) and dTDP-d-glucose-4,6-dehydratase (RmlB). The enzymes catalyzing the last three synthesis reactions have not been characterized biochemically so far. These steps include an isomerase, a transaminase, and a transacetylase. We identified all five genes involved by chromosome walking in the Gram-positive organism A. thermoaerophilus L420-91T and overexpressed the three new enzymes heterologously in Escherichia coli. The activities of these enzymes were monitored by reverse phase high performance liquid chromatography, and the intermediate products formed were characterized by 1H and 13C nuclear magnetic resonance spectroscopy analysis. Alignment of the newly identified proteins with known sequences revealed that the elucidated pathway in this Gram-positive organism may also be valid in the biosynthesis of the O-antigen of lipopolysaccharides of Gram negative organisms. The key enzyme in the biosynthesis of dTDP-d-Fucp3NAc has been identified as an isomerase, which converts the 4-keto educt into the 3-keto product, with concomitant epimerization at C-4 to produce a 6-deoxy-d-xylo configuration. This is the first report of the functional characterization of the biosynthesis of dTDP-d-Fucp3NAc and description of a novel type of isomerase capable of synthesizing dTDP-6-deoxy-d-xylohex-3-ulose from dTDP-6-deoxy-d xylohex-4-ulose. PMID- 12740381 TI - Protein kinase A-anchoring protein AKAP95 interacts with MCM2, a regulator of DNA replication. AB - Protein kinase A (PKA)-anchoring protein AKAP95 is localized to the nucleus in interphase, where it primarily associates with the nuclear matrix. A yeast two hybrid screen for AKAP95 interaction partners identified the minichromosome maintenance (MCM) 2 protein, a component of the pre-replication complex. AKAP95 MCM2 interaction was mapped to residues 1-195 of AKAP95 and corroborated by glutathione S-transferase precipitation and immunoprecipitation from chromatin. Disruption of AKAP95-MCM2 interaction with an AKAP95-(1-195) peptide within HeLa cell nuclei abolishes initiation of DNA replication in G1 phase and the elongation phase of replication in vitro without affecting global nuclear organization or import. Disruption of the C-terminal zinc finger of AKAP95 reduces efficiency of replication initiation. Disruption of the PKA-binding domain does not impair replication in G1- or S-phase nuclei, whereas a PKA inhibitor affects the initiation but not the elongation phase of replication. Depleting AKAP95 from nuclei partially depletes MCM2 and abolishes replication. Recombinant AKAP95 restores intranuclear MCM2 and replication in a dose-dependent manner. Our results suggest a role of AKAP95 in DNA replication by providing a scaffold for MCM2. PMID- 12740382 TI - Calreticulin promotes folding/dimerization of human lipoprotein lipase expressed in insect cells (sf21). AB - Lipoprotein lipase (LPL) is a non-covalent, homodimeric, N-glycosylated enzyme important for metabolism of blood lipids. LPL is regulated by yet unknown post translational events affecting the levels of active dimers. On co-expression of LPL with human molecular chaperones, we found that calreticulin had the most pronounced effects on LPL activity, but calnexin was also effective. Calreticulin caused a 9-fold increase in active LPL, amounting to about 50% of the expressed LPL protein. The total expression of LPL protein was increased less than 20%, and the secretion rates for active and inactive LPL were not significantly changed by the chaperone. Thus, the main effect was an increased specific activity of LPL both in cells and media. Chromatography on heparin-Sepharose and sucrose density gradient centrifugation demonstrated that most of the inactive LPL was monomeric and that calreticulin promoted formation of active dimers. Higher oligomers of inactive LPL were present in cell extracts, but only monomers and dimers were secreted to the medium. Interaction between LPL and calreticulin was demonstrated, and the effect of the chaperone was prevented by castanospermine, an inhibitor of N-glycan glucose trimming. Our data indicate an important role of endoplasmic reticulum-based chaperones for the folding/dimerization of LPL. PMID- 12740383 TI - The Wnt signaling inhibitor dickkopf-1 is required for reentry into the cell cycle of human adult stem cells from bone marrow. AB - Adult human mesenchymal stem cells from bone marrow stroma (hMSCs) differentiate into numerous mesenchymal tissue lineages and are attractive candidates for cell and gene therapy. When early passage hMSCs are plated or replated at low density, the cultures display a lag phase of 3-5 days, a phase of rapid exponential growth, and then enter a stationary phase without the cultures reaching confluence. We found that as the cultures leave the lag phase, they secrete high levels of dickkopf-1 (Dkk-1), an inhibitor of the canonical Wnt signaling pathway. The addition of recombinant Dkk-1 toward the end of the lag period increased proliferation and decreased the cellular concentration of beta-catenin. The addition of antibodies to Dkk-1 in the early log phase decreased proliferation. Also, expression of Dkk-1 in hMSCs decreased during cell cycle arrest induced by serum starvation. The results indicated that high levels of Dkk 1 allow the cells to reenter the cell cycle by inhibiting the canonical Wnt/beta catenin signaling pathway. Since antibodies to Dkk-1 also increased the lag phase of an osteosarcoma line that expressed the gene, Dkk-1 may have a similar role in some other cell systems. PMID- 12740384 TI - Insights into the structural basis for zinc inhibition of the glycine receptor. AB - Histidines 107 and 109 in the glycine receptor (GlyR) alpha1 subunit have previously been identified as determinants of the inhibitory zinc-binding site. Based on modeling of the GlyR alpha1 subunit extracellular domain by homology to the acetylcholine-binding protein crystal structure, we hypothesized that inhibitory zinc is bound within the vestibule lumen at subunit interfaces, where it is ligated by His107 from one subunit and His109 from an adjacent subunit. This was tested by co-expressing alpha1 subunits containing the H107A mutation with alpha1 subunits containing the H109A mutation. Although sensitivity to zinc inhibition is markedly reduced when either mutation is individually incorporated into all five subunits, the GlyRs formed by the co-expression of H107A mutant subunits with H109A mutant subunits exhibited an inhibitory zinc sensitivity similar to that of the wild type alpha1 homomeric GlyR. This constitutes strong evidence that inhibitory zinc is coordinated at the interface between adjacent alpha1 subunits. No evidence was found for beta subunit involvement in the coordination of inhibitory zinc, indicating that a maximum of two zinc-binding sites per alpha1beta receptor is sufficient for maximal zinc inhibition. Our data also show that two zinc-binding sites are sufficient for significant inhibition of alpha1 homomers. The binding of zinc at the interface between adjacent alpha1 subunits could restrict intersubunit movements, providing a feasible mechanism for the inhibition of channel activation by zinc. PMID- 12740385 TI - Lysophosphatidylcholine acts as an anti-hemostatic molecule in the saliva of the blood-sucking bug Rhodnius prolixus. AB - Blood-sucking arthropods possess a variety of anti-hemostatic factors in their salivary glands to maintain blood fluidity during feeding. In this work we demonstrate the anti-hemostatic properties of lysophosphatidylcholine (lysoPC) isolated from the salivary glands of Rhodnius prolixus. First, we examined salivary glands of fourth and fifth instar nymphs for their phospholipid composition. The lumen displayed an accumulation of its phospholipid content, mainly phosphatidylcholine and lysoPC, with a 6-fold increase for the latter. To determine the presence of phospholipids in the saliva, fourth instar nymphs were fed with a32P-enriched blood meal. After 28 days their saliva was collected and subjected to lipid extraction, thin-layer chromatography, and autoradiography. The results showed the presence in the saliva of the same phospholipids present in the lumen. We then examined possible biological roles of these phospholipids when compared with other known effects of lysoPC. The luminal lipid extract and purified lysoPC from the lumen and saliva were tested for inhibition of washed rabbit platelets' aggregation induced by alpha-thrombin and platelet-activating factor. Both the luminal lipid extract and salivary lysoPC showed an increasing inhibition of aggregation, which correlated with the response of the platelets to standard lysoPC (up to 13 microg/ml). Next, salivary lysoPC was incubated with porcine arterial endothelial cells for 24 h. After incubation, culture medium was assayed for nitric oxide and showed increased nitric oxide production, similar to control cells exposed to standard lysoPC (up to 20 microg/ml). Together these data demonstrate the presence of lysoPC in the saliva of Rhodnius prolixus and its potential anti-hemostatic activities. PMID- 12740386 TI - Mechanism of ribosomal p70S6 kinase activation by granulocyte macrophage colony stimulating factor in neutrophils: cooperation of a MEK-related, THR421/SER424 kinase and a rapamycin-sensitive, m-TOR-related THR389 kinase. AB - We report here for the first time the detection of the ribosomal p70S6 kinase (p70S6K) in a hematopoietic cell, the neutrophil, and the stimulation of its enzymatic activity by granulocyte macrophage colony-stimulating factor (GM-CSF). GM-CSF modified the Vmax of the enzyme (from 7.2 to 20.5 pmol/min/mg) and induced a time- and dose-dependent phosphorylation on p70S6K residues Thr389 and Thr421/Ser424. The immunosuppressant macrolide rapamycin caused either a decrease in intensity of phospho-Thr389 bands in Western blots, or as a downshift in the relative mobility of phospho-Thr421/Ser424 bands (consistent with the loss of phosphate), but not both simultaneously. The immunosuppressant FK506 failed to inhibit p70S6K activation, but was able to rescue the rapamycin-induced downshift, pointing to a role for the mammalian target of rapamycin (mTOR) kinase. Rapamycin also caused an inhibition (IC50 0.2 nm) of the in vitro enzymatic activity of p70S6K. However, the inhibition of activity was not complete, but only a 40-50%, indicating that neutrophil p70S6K activity has a rapamycin-resistant component. This component was totally inhibited by pre incubating the cells with the mitogen-activated protein kinase (MAPK) kinase (MEK) inhibitor PD-98059 prior to treatment with rapamycin. This indicated that a kinase from the MEK/MAPK pathway also plays a role in p70S6K activation. Thus, GM CSF causes the dual activation of a rapamycin-resistant, MAPK-related kinase, that targets Thr421/Ser424 S6K phosphorylation, and a rapamycin-sensitive, mTOR related kinase, that targets Thr389, both of which are needed in cooperation to achieve full activation of neutrophil p70S6K. PMID- 12740387 TI - Central role of a serine phosphorylation site within duck hepatitis B virus core protein for capsid trafficking and genome release. AB - Viral nucleocapsids compartmentalize and protect viral genomes during assembly while they mediate targeted genome release during viral infection. This dual role of the capsid in the viral life cycle must be tightly regulated to ensure efficient virus spread. Here, we used the duck hepatitis B virus (DHBV) infection model to analyze the effects of capsid phosphorylation and hydrogen bond formation. The potential key phosphorylation site at serine 245 within the core protein, the building block of DHBV capsids, was substituted by alanine (S245A), aspartic acid (S245D) and asparagine (S245N), respectively. Mutant capsids were analyzed for replication competence, stability, nuclear transport, and infectivity. All mutants formed DHBV DNA-containing nucleocapsids. Wild-type and S245N but not S245A and S245D fully protected capsid-associated mature viral DNA from nuclease action. A negative ionic charge as contributed by phosphorylated serine or aspartic acid-supported nuclear localization of the viral capsid and generation of nuclear superhelical DNA. Finally, wild-type and S245D but not S245N virions were infectious in primary duck hepatocytes. These results suggest that hydrogen bonds formed by non-phosphorylated serine 245 stabilize the quarterny structure of DHBV nucleocapsids during viral assembly, while serine phosphorylation plays an important role in nuclear targeting and DNA release from capsids during viral infection. PMID- 12740388 TI - Conservation in the mechanism of Nedd8 activation by the human AppBp1-Uba3 heterodimer. AB - Human Nedd8-activating enzyme AppBp1-Uba3 was purified to apparent homogeneity from erythrocytes. In the presence of [2,8-3H]ATP and 125I-Nedd8, heterodimer rapidly forms a stable stoichiometric ternary complex composed of tightly bound Nedd8 [3H]adenylate and Uba3-125I-Nedd8 thiol ester. Isotope exchange kinetics show that the heterodimer follows a pseudo-ordered mechanism with ATP the leading and Nedd8 the trailing substrate. Human AppBp1-Uba3 follows hyperbolic kinetics for HsUbc12 transthiolation with 125I-Nedd8 (kcat = 3.5 +/- 0.2 s-1), yielding Km values for ATP (103 +/- 12 microm), 125I-Nedd8 (0.95 +/- 0.18 microm), and HsUbc12 (43 +/- 13 nm) similar to those for ubiquitin activation by Uba1. Wild type 125I-ubiquitin fails to support AppBp1-Uba3 catalyzed activation or HsUbc12 transthiolation. However, modest inhibition of 125I-Nedd8 ternary complex formation by unlabeled ubiquitin suggests a Kd > 300 microm for ubiquitin. Alanine 72 of Nedd8 is a critical specificity determinant for AppBp1-Uba3 binding because 125I-UbR72L undergoes heterodimer-catalyzed hyperbolic HsUbc12 transthiolation and yields Km = 20 +/- 9 microm and kcat = 0.9 +/- 0.3 s-1. These observations demonstrate remarkable conservation in the mechanism of AppBp1-Uba3 that mirrors its sequence conservation with the Uba1 ubiquitin-activating enzyme. PMID- 12740389 TI - Sumoylation of Smad4, the common Smad mediator of transforming growth factor-beta family signaling. AB - Transforming growth factor-beta (TGF-beta) and TGF-beta-related factors regulate cell growth, differentiation, and apoptosis, and play key roles in normal development and tumorigenesis. TGF-beta family-induced changes in gene expression are mediated by serine/threonine kinase receptors at the cell surface and Smads as intracellular effectors. Receptor-activated Smads combine with a common Smad4 to translocate into the nucleus where they cooperate with other transcription factors to activate or repress transcription. The activities of the receptor activated Smads are controlled by post-translational modifications such as phosphorylation and ubiquitylation. Here we show that Smad4 is modified by sumoylation. Sumoylation of Smad4 was enhanced by the conjugating enzyme Ubc9 and members of the PIAS family of SUMO ligases. A major sumoylation site in Smad4 was localized to Lys-159 in its linker segment with an additional site at Lys-113 in the MH-1 domain. Increased sumoylation in the presence of the PIASy E3 ligase correlated with targeting of Smad4 to subnuclear speckles that contain SUMO-1 and PIASy. Replacement of lysines 159 and 113 by arginines or increased sumoylation enhanced the stability of Smad4, and transcription in mammalian cells and Xenopus embryos. These observations suggest a role for Smad4 sumoylation in the regulation of TGF-beta signaling through Smads. PMID- 12740390 TI - Myosin IIb is unconventionally conventional. AB - Members of the myosin II class of molecular motors have been referred to as "conventional," a term used to describe their ability to form thick filaments, their low duty ratio, the ability of individual motor-containing "heads" to operate independently of each other, and their rate-limiting phosphate release. These features ensure that those motors that have completed their power stroke dissociate rapidly enough to prevent them from interfering with those motors that are beginning theirs. However, in this study, we demonstrate that myosin IIB, a cytoplasmic myosin II particularly enriched in the central nervous system and cardiac tissue, has a number of features that it shares instead with "unconventional" myosin isoforms, including myosins V and VI. These include a high duty ratio, rate-limiting ADP release, and high ADP affinity. These features imply that myosin IIB serves a set of physiologic needs different from those served by its more conventional myosin II counterparts, and this work provides a plausible basis for explaining the physiologic role of this unconventionally conventional myosin. PMID- 12740391 TI - Synergistic promotion of c-Src activation and cell migration by Cas and AND 34/BCAR3. AB - The adapter molecule p130Cas (Cas) plays a role in cellular processes such as proliferation, survival, cell adhesion, and migration. The ability of Cas to promote migration has been shown to be dependent upon its carboxyl terminus, which contains a bipartite binding site for the protein tyrosine kinase c-Src (Src). The association between Src and Cas enhances Src kinase activity, and like Cas, Src plays an important role in cell proliferation and migration. In this study, we show that Src and Cas function cooperatively to promote cell migration in a manner that depends upon kinase-active Src. Another carboxyl-terminal binding partner of Cas, AND-34/BCAR3 (AND-34), functions synergistically with Cas to enhance Src activation and cell migration. The carboxyl-terminal guanine nucleotide exchange factor domain of AND-34, as well as the activity of its putative target Rap1, contribute to these events. A mechanism through which AND 34 may regulate Cas-dependent cell migration is suggested by the finding that Cas becomes redistributed from focal adhesions to lamellipodia located at the leading edge of AND-34 overexpressing cells. These data thus provide insight into how Cas and AND-34 may function together to stimulate Src signaling pathways and promote cell migration. PMID- 12740392 TI - Tage4/Nectin-like molecule-5 heterophilically trans-interacts with cell adhesion molecule Nectin-3 and enhances cell migration. AB - Malignant transformation of cells causes disruption of cell-cell adhesion, enhancement of cell motility, and invasion into surrounding tissues. Nectins have both homophilic and heterophilic cell-cell adhesion activities and organize adherens junctions in cooperation with cadherins. We examined here whether Tage4, which was originally identified to be a gene overexpressed in colon carcinoma and has a domain structure similar to those of nectins, is involved in cell adhesion and/or migration. Tage4 heterophilically trans-interacted with nectin-3, but not homophilically with Tage4. Expression of Tage4 was markedly elevated in NIH3T3 cells transformed by an oncogenic Ki-Ras (V12Ras-NIH3T3 cells) as compared with that of wild-type NIH3T3 cells. trans-Interaction of Tage4 with nectin-3 enhanced motility of V12Ras-NIH3T3 cells. Tage4 did not bind afadin, a nectin- and actin filament-binding protein that connects nectins to the actin cytoskeleton and cadherins through catenins. Thus, Tage4 heterophilically trans-interacts with nectin-3 and regulates cell migration. Tage4 is tentatively re-named here nectin like molecule-5 (necl-5) on the basis of its function and domain structure similar to those of nectins. PMID- 12740393 TI - Neck length and processivity of myosin V. AB - Myosin V is an unconventional myosin that transports cargo such as vesicles, melanosomes, or mRNA on actin filaments. It is a two-headed myosin with an unusually long neck that has six IQ motifs complexed with calmodulin. In vitro studies have shown that myosin V moves processively on actin, taking multiple 36 nm steps that coincide with the helical repeat of actin. This allows the molecule to "walk" across the top of an actin filament, a feature necessary for moving large vesicles along an actin filament bound to the cytoskeleton. The extended neck length of the two heads is thought to be critical for taking 36-nm steps for processive movements. To test this hypothesis we have expressed myosin V heavy meromyosin-like fragments containing 6IQ motifs, as well as ones that shorten (2IQ, 4IQ) or lengthen (8IQ) the neck region or alter the spacing between 3rd and 4th IQ motifs. The step size was proportional to neck length for the 2IQ, 4IQ, 6IQ, and 8IQ molecules, but the molecule with the altered spacing took shorter than expected steps. Total internal reflection fluorescence microscopy was used to determine whether the heavy meromyosin IQ molecules were capable of processive movements on actin. At saturating ATP concentrations, all molecules except for the 2IQ mutant moved processively on actin. When the ATP concentration was lowered to 10 microm or less, the 2IQ mutant demonstrated some processive movements but with reduced run lengths compared with the other mutants. Its weak processivity was also confirmed by actin landing assays. PMID- 12740394 TI - Post-translational modifications of three members of the human MAP1LC3 family and detection of a novel type of modification for MAP1LC3B. AB - The molecular machinery required for autophagy is highly conserved in all eukaryotes as seen by the high degree of conservation of proteins involved in the formation of the autophagosome membranes. Recently, both yeast Apg8p and its rat homologue Map1lc3 were identified as essential constituents of autophagosome membrane as a processed form. In addition, both the yeast and human proteins exist in two modified forms produced by a series of post-translational modifications including a critical C-terminal cleavage after a conserved Gly residue, and the smaller processed form is associated with the autophagosome membranes. Herein, we report the identification and characterization of three human orthologs of the rat Map1LC3, named MAP1LC3A, MAP1LC3B, and MAP1LC3C. We show that the three isoforms of human MAP1LC3 exhibit distinct expression patterns in different human tissues. Importantly, we found that the three isoforms of MAP1LC3 differ in their post-translation modifications. Although MAP1LC3A and MAP1LC3C are produced by the proteolytic cleavage after the conserved C-terminal Gly residue, like their rat counterpart, MAP1LC3B does not undergo C-terminal cleavage and exists in a single modified form. The essential site for the distinct post-translation modification of MAP1LC3B is Lys-122 rather than the conserved Gly-120. Subcellular localization by cell fractionation and immunofluorescence revealed that three human isoforms are associated with membranes involved in the autophagic pathway. These results revealed different regulation of the three human isoforms of MAP1LC3 and implicate that the three isoforms may have different physiological functions. PMID- 12740396 TI - Structural analysis of the DNA-binding domain of the Erwinia amylovora RcsB protein and its interaction with the RcsAB box. AB - The transcriptional regulator RcsB interacts with other coactivators to control the expression of biosynthetic operons in enterobacteria. While in a heterodimer complex with the regulator RcsA the RcsAB box consensus is recognized, DNA binding sites for RcsB without RcsA have also been identified. The conformation of RcsB might therefore be modulated upon interaction with various coactivators, resulting in the recognition of different DNA targets. We report the solution structure of the C-terminal DNA-binding domain of the RcsB protein from Erwinia amylovora spanning amino acid residues 129-215 solved by heteronuclear magnetic resonance (NMR) spectroscopy. The C-terminal domain is composed of four alpha helices where two central helices form a helix-turn-helix motif similar to the structures of the regulatory proteins GerE, NarL, and TraR. Amino acid residues involved in the RcsA independent DNA binding of RcsB were identified by titration studies with a RcsAB box consensus fragment. Data obtained from NMR spectroscopy together with surface plasmon resonance measurements demonstrate that the RcsAB box is specifically recognized by the RcsAB heterodimer as well as by RcsB alone. However, the binding constant of RcsB alone at target promoters from Escherichia coli, E. amylovora, and Pantoea stewartii was approximately 1 order of magnitude higher compared with that of the RcsAB heterodimer. We present evidence that the obvious role of RcsA is not to alter the DNA binding specificity of RcsB but to stabilize RcsB-DNA complexes. PMID- 12740397 TI - Nitrergic prejunctional inhibition of purinergic neuromuscular transmission in the hamster proximal colon. AB - Neurogenic ATP and nitric oxide (NO) may play important roles in the physiological control of gastrointestinal motility. However, the interplay between purinergic and nitrergic neurons in mediating the inhibitory neurotransmission remains uncertain. This study investigated whether neurogenic NO modulates the purinergic transmission to circular smooth muscles of the hamster proximal colon. Electrical activity was recorded from circular muscle cells of the hamster proximal colon by using the microelectrode technique. Intramural nerve stimulation with a single pulse evoked a fast purinergic inhibitory junction potential (IJP) followed by a slow nitrergic IJP. The purinergic component of the second IJP evoked by paired stimulus pulses at pulse intervals between 1 and 3 s became smaller than that of the first IJP. This purinergic IJP depression could be observed at pulse intervals <3 s, but not at longer ones, and failed to occur in the presence of NO synthase inhibitor. Exogenous NO (0.3-1 microM), at which no hyperpolarization is produced, inhibited purinergic IJPs, without altering the nitrergic IJP and exogenously applied ATP induced hyperpolarization. In the presence of both purinoceptor antagonist and nitric oxide synthase (NOS) inhibitor, intramural nerve stimulation with 5 pulses at 20 Hz evoked vasoactive intestinal peptide (VIP)-associated IJPs, suggesting that VIP component may be masked in the IJPs of the hamster proximal colon. Our results suggest that neurogenic NO may modulate the purinergic transmission to circular smooth muscles of the hamster proximal colon via a prejunctional mechanism. In addition, VIP may be involved in the neurotransmitter in the hamster proximal colon. PMID- 12740395 TI - M phase phosphoprotein 1 is a human plus-end-directed kinesin-related protein required for cytokinesis. AB - The human M phase phosphoprotein 1 (MPP1), previously identified through a screening of a subset of proteins specifically phosphorylated at the G2/M transition (Matsumoto-Taniura, N., Pirollet, F., Monroe, R., Gerace, L., and Westendorf, J. M. (1996) Mol. Biol. Cell 7, 1455-1469), is characterized as a plus-end-directed kinesin-related protein. Recombinant MPP1 exhibits in vitro microtubule-binding and microtubule-bundling properties as well as microtubule stimulated ATPase activity. In gliding experiments using polarity-marked microtubules, MPP1 is a slow molecular motor that moves toward the microtubule plus-end at a 0.07 microm/s speed. In cycling cells, MPP1 localizes mainly to the nuclei in interphase. During mitosis, MPP1 is diffuse throughout the cytoplasm in metaphase and subsequently localizes to the midzone to further concentrate on the midbody. MPP1 suppression by RNA interference induces failure of cell division late in cytokinesis. We conclude that MPP1 is a new mitotic molecular motor required for completion of cytokinesis. PMID- 12740399 TI - Asymmetry in the pitch control system of the lamprey caused by a unilateral labyrinthectomy. AB - A postural control system in the lamprey is driven by vestibular input and maintains a definite orientation of the animal during swimming. After a unilateral labyrinthectomy (UL), the lamprey continuously rolls toward the damaged side. Important elements of the postural network are the reticulospinal (RS) neurons that are driven by vestibular input and transmit commands for postural corrections to the spinal cord. We characterized the effect of UL on vestibular responses in RS neurons elicited by rotation of the animal in the pitch plane. The activity of RS neurons was recorded from their axons in the spinal cord before and after UL. The neurons can be classified into the Up and Down groups activated preferentially with nose-up or nose-down rotation, respectively. After UL, vestibular responses in the group Up changed only slightly on the damaged side and disappeared almost completely on the opposite side. In the group Down, responses on both sides persisted after UL. These results indicate that the left and right subgroups of the group Up neurons receive excitatory input mainly from the contralateral labyrinth. In contrast, the group Down neurons receive excitatory input from both labyrinths. We conclude that the UL-induced changes in vestibular responses to pitch tilt will disturb the normal activity of the postural control system. The UL-induced asymmetry in the bilateral activity of the group Up neurons seems to be an important factor contributing to the loss of equilibrium in UL animals and to their rotation during swimming. PMID- 12740398 TI - Protein synthesis inhibition blocks the late-phase LTP of C-fiber evoked field potentials in rat spinal dorsal horn. AB - Previous studies have demonstrated that in the hippocampus the maintenance of long-term potentiation (LTP) requires de novo protein synthesis. To investigate the role of protein synthesis in the maintenance of LTP of C-fiber evoked field potentials in spinal dorsal horn, which may be relevant to hyperalgesia, protein synthesis inhibitor (either cycloheximide or anisomycin) was applied locally to the recording segments of spinal cord in anesthetized rats, 30 min prior to tetanic stimulation to the sciatic nerve. We found that both cycloheximide and anisomycin selectively inhibited late-phase maintenance of the spinal LTP but affected neither LTP induction nor baseline responses of C-fiber evoked field potentials. In the presence of cycloheximide, LTP of C-fiber evoked field potentials was 281.5 +/- 16.5% (n = 6) of baseline 1 h after tetanic stimulation and the potentiation significantly decreased to 235.5 +/- 18.5% at 145 min after tetanic stimulation (P < 0.05). Afterward, LTP of C-fiber evoked field potentials decreased continuously and at 270 min after tetanic stimulation reached 130.8 +/- 18.0%, which was no longer different from baseline (P > 0.05). Spinal application of anisomycin at 30 min before tetanic stimulation yielded similar results (n = 6). These results suggest that protein synthesis may be crucial for the late phase maintenance of LTP of C-fiber evoked field potentials in spinal dorsal horn. PMID- 12740400 TI - Kappa opioid receptor activation in the nucleus accumbens inhibits glutamate and GABA release through different mechanisms. AB - Through their actions in the nucleus accumbens (NAc), kappa opioid (KOP) receptors and their endogenous ligand, dynorphin, modify behaviors associated with the administration of drugs of abuse and are regulated by exposure to such drugs. Despite their demonstrated behavioral significance, the synaptic actions of KOP receptor ligands in the NAc are not clearly understood. Using whole-cell voltage-clamp recordings of NAc medium spiny neurons, we have found that, in addition to suppressing glutamate release, the KOP receptor agonist also inhibits GABA release. Interestingly, the mechanism of inhibition of the release of glutamate differs from that controlling GABA. reduces the frequency of Ca(2+) independent miniature excitatory postsynaptic currents, but not miniature inhibitory postsynaptic currents. Furthermore, while the inhibition of GABAergic transmission is blocked by the N-type Ca(2+) channel blocker omega-CgTx, the inhibition of excitatory glutamatergic transmission by is unaffected by N-type Ca(2+) channel blockade. These results indicate that KOP receptor activation inhibits GABA release by reducing Ca(2+) influx, but inhibits glutamate release at a step downstream of Ca(2+) entry. PMID- 12740401 TI - Effects of noise on the spike timing precision of retinal ganglion cells. AB - Information in a spike train is limited by variability in the spike timing. This variability is caused by noise from several sources including synapses and membrane channels; but how deleterious each noise source is and how they affect spike train coding is unknown. Combining physiology and a multicompartment model, we studied the effect of synaptic input noise and voltage-gated channel noise on spike train reliability for a mammalian ganglion cell. For tonic stimuli, the SD of the interspike intervals increased supralinearly with increasing interspike interval. When the cell was driven by current injection, voltage-gated channel noise and background synaptic noise caused fluctuations in the interspike interval of comparable amplitude. Spikes initiated on the dendrites could cause additional spike timing fluctuations. For transient stimuli, synaptic noise was dominant and spontaneous background activity strongly increased fluctuations in spike timing but decreased the latency of the first spike. PMID- 12740402 TI - Interneuronal projections to identified cilia-activating pedal neurons in Hermissenda. AB - Neural networks have been shown to support the generation of more than one behavioral motor act. In the nudibranch mollusk Hermissenda, Pavlovian conditioning results in light, the conditioned stimulus (CS), evoking both inhibition of locomotion and foot contraction. The synaptic organization of the eyes and optic ganglion is well documented; however, the characterization of the neural network mediating visually modulated behaviors is incomplete. We have now characterized synaptic connections between identified photoreceptors and a newly identified interneuron (II(b)), identified synaptic projections from type I and type II interneurons to an inhibitory interneuron (III(i)) and to two newly identified pedal neurons, VP1 and VP2. Here we show that VP1 activates ciliary movement on the anterior foot and VP2 innervates the anterior foot and ventral tentacle. Stimulation of the photoreceptors with light produced two effects on the activity of VP1 and VP2. First, light inhibits type I(i) and II(i) interneurons and disinhibits VP1 and VP2. Depolarization of type II(e) interneurons also disinhibits VP1 and VP2. Second, the light-elicited depolarization and increased tonic activity of VP1 and VP2 is produced by excitatory synaptic input from ipsilateral and contralateral type II(b) interneurons. Pedal neurons VP1 and VP2 receive similar synaptic input from type I, II, and III(i) interneurons; this is in agreement with previous research showing that the visual pathway influences both ciliary locomotion and foot movement. The organization of the visual system in Hermissenda provides for the expression of cellular and synaptic plasticity supporting learning without altering the networks ability to carry out the requirements for normal visual processing. PMID- 12740403 TI - Simulations to derive membrane resistivity in three phenotypes of guinea pig sympathetic postganglionic neuron. AB - The electrotonic behavior of three phenotypes of sympathetic postganglionic neuron has been analyzed to assess whether their distinct cell input capacitances simply reflect differences in morphology. Because the distribution of membrane properties over the soma and dendrites is unknown, compartmental models incorporating cell morphology were used to simulate hyperpolarizing responses to small current steps. Neurons were classified as phasic (Ph), tonic (T), or long afterhyperpolarizing (LAH) by their discharge pattern to threshold depolarizing current steps and filled with biocytin to determine their morphology. Responses were simulated in models with the average morphology of each cell class using the program NEURON. Specific membrane resistivity, R(m), was derived in each model. Fits were acceptable when specific membrane capacitance, C(m), and specific resistivity of the axoplasm, R(i,) were varied within realistic limits and when underestimation of membrane area due to surface irregularities was accounted for. In all models with uniform R(m), solutions for R(m) that were the same for all classes could not be found unless C(m) or R(i) were different for each class, which seems unrealistic. Incorporation of a small somatic shunt conductance yielded values for R(m) for each class close to those derived assuming isopotentiality (R(m) approximately 40, 27, and 15 k omega cm(2) for T, Ph, and LAH neurons, respectively). It is concluded that R(m) is distinct between neuron classes. Because Ph and LAH neurons relay selected preganglionic inputs directly, R(m) generally affects function only in T neurons that integrate multiple subthreshold inputs and are modulated by peptidergic transmitters. PMID- 12740404 TI - Regulation of backpropagating action potentials in mitral cell lateral dendrites by A-type potassium currents. AB - Dendrodendritic synapses, distributed along mitral cell lateral dendrites, provide powerful and extensive inhibition in the olfactory bulb. Activation of inhibition depends on effective penetration of action potentials into dendrites. Although action potentials backpropagate with remarkable fidelity in apical dendrites, this issue is controversial for lateral dendrites. We used paired somatic and dendritic recordings to measure action potentials in proximal dendritic segments (0-200 microm from soma) and action potential-generated calcium transients to monitor activity in distal dendritic segments (200-600 microm from soma). Somatically elicited action potentials were attenuated in proximal lateral dendrites. The attenuation was not due to impaired access resistance in dendrites or to basal synaptic activity. However, a single somatically elicited action potential was sufficient to evoke a calcium transient throughout the lateral dendrite, suggesting that action potentials reach distal dendritic compartments. Block of A-type potassium channels (I(A)) with 4 aminopyridine (10 mM) prevented action potential attenuation in direct recordings and significantly increased dendritic calcium transients, particularly in distal dendritic compartments. Our results suggest that I(A) may regulate inhibition in the olfactory bulb by controlling action potential amplitudes in lateral dendrites. PMID- 12740405 TI - Fast Ca2+-induced potentiation of heat-activated ionic currents requires cAMP/PKA signaling and functional AKAP anchoring. AB - Calcium influx and the resulting increase in intracellular calcium concentration ([Ca(2+)](i)) can induce enhanced sensitivity to temperature increases in nociceptive neurons. This sensitization accounts for heat hyperalgesia that is regularly observed following the activation of excitatory inward currents by pain producing mediators. Here we show that rat sensory neurons express calcium dependent adenylyl cyclases (AC) using RT-PCR and nonradioactive in situ hybridization. Ionomycin-induced rises in [Ca(2+)](i)-activated calcium-dependent AC and caused translocation of catalytic protein kinase A subunit. Elevation of [Ca(2+)](i) finally resulted in a significant potentiation of heat-activated currents and a drop in heat threshold. This was not prevented in the presence of suramin that nonspecifically uncouples G protein-dependent receptors. The sensitization was, however, inhibited when the specific PKA antagonist PKI(14-22) was added to the pipette solution or when PKA coupling to A kinase anchoring protein (AKAP) was disrupted with InCELLect StHt-31 uncoupling peptide. The results show that heat sensitization in nociceptive neurons can be induced by increases in [Ca(2+)](i) and requires PKA that is functionally coupled to the heat transducer, mostly likely vanilloid receptor VR-1. This calcium-dependent pathway can account for the sensitizing properties of many excitatory mediators that activate cationic membrane currents. PMID- 12740406 TI - Spatial memory following shifts of gaze. I. Saccades to memorized world-fixed and gaze-fixed targets. AB - During a shift of gaze, an object can move along with gaze or stay fixed in the world. To examine the effect of an object's reference frame on spatial working memory, we trained monkeys to memorize locations of visual stimuli as either fixed in the world or fixed to gaze. Each trial consisted of an initial reference frame instruction, followed by a peripheral visual flash, a memory-period gaze shift, and finally a memory-guided saccade to the location consistent with the instructed reference frame. The memory-period gaze shift was either rapid (a saccade) or slow (smooth pursuit or whole body rotation). This design allowed a comparison of memory-guided saccade performance under various conditions. Our data indicate that after a rotation or smooth-pursuit eye movement, saccades to memorized world-fixed targets are more variable than saccades to memorized gaze fixed targets. In contrast, memory-guided saccades to world- and gaze-fixed targets are equally variable following a visually guided saccade. Across all conditions, accuracy, latency, and main sequence characteristics of memory-guided saccades are not influenced by the target's reference frame. Memory-guided saccades are, however, more accurate after fast compared with slow gaze shifts. These results are most consistent with an eye-centered representational system for storing the spatial locations of memorized objects but suggest that the visual system may engage different mechanisms to update the stored signal depending on how gaze is shifted. PMID- 12740407 TI - Both electrical and chemical synapses mediate fast network oscillations in the olfactory bulb. AB - Odor perception depends on a constellation of molecular, cellular, and network interactions in olfactory brain areas. Recently, there has been better understanding of the cellular and molecular mechanisms underlying the odor responses of neurons in the olfactory epithelium, the first-order olfactory area. In higher order sensory areas, synchronized activity in networks of neurons is known to be a prominent feature of odor processing. The perception and discrimination of odorants is associated with fast (20-70 Hz) electroencephalographic oscillations. The cellular mechanisms underlying these fast network oscillations have not been defined. In this study, we show that synchronous fast oscillations can be evoked by brief electrical stimulation in the rat olfactory bulb in vitro, partially mimicking the natural response of this brain region to sensory input. Stimulation induces periodic inhibitory synaptic potentials in mitral cells and prolonged spiking in GABAergic granule cells. Repeated stimulation leads to the persistent enhancement in both granule cell activity and mitral cell inhibition. Prominent oscillations in field recordings indicate that stimulation induces high-frequency activity throughout networks of olfactory bulb neurons. Network synchronization results from chemical and electrical synaptic interactions since both glutamate-receptor antagonists and gap junction inhibitors block oscillatory intracellular and field responses. Our results demonstrate that the olfactory bulb can generate fast oscillations autonomously through the persistent activation of networks of inhibitory interneurons. These local circuit interactions may be critically involved in odor processing in vivo. PMID- 12740408 TI - Cortical projection of peripheral vestibular signaling. AB - The cerebral projection of vestibular signaling was studied by using PET with a special differential experimental protocol. Caloric vestibular stimulation (CVS) induced regional cerebral blood flow (rCBF) changes were investigated in two populations. Butanol perfusion scans were carried out on six healthy volunteers and on six patients following the removal of tumors from the right cerebello pontine angle. The complete loss of the vestibular function postoperatively allowed a comparison of the rCBF changes in the populations with or without this input and offered a promising functional approach whereby to delineate the cortical region most responsive to pure vestibular input. The activations by left sided and right-sided CVS were determined for both the healthy volunteers and the patient population. Statistical analysis of the data obtained following left sided CVS did not reveal any cerebral region for which there was a significant difference in CVS-induced response by these two populations. In the case of right sided CVS, however, the statistical comparison of the CVS-related responses demonstrated a single contralateral area characterized by a significantly different degree of response. This cortical area corresponds to part of the cortical region described recently which can be activated by both CVS and neck vibration. It appears to be anatomically identical to the aggregate of the somatosensory area SII and the retroinsular cortex described in primates, a region identified by other investigators as an analog of the parietoinsular vestibular cortex. PMID- 12740409 TI - Cannabinoid receptor activation differentially modulates ion channels in photoreceptors of the tiger salamander. AB - Cannabinoid CB1 receptors have been detected in retinas of numerous species, with prominent labeling in photoreceptor terminals of the chick and monkey. CB1 labeling is well-conserved across species, suggesting that CB1 receptors might also be present in photoreceptors of the tiger salamander. Synaptic transmission in vertebrate photoreceptors is mediated by L-type calcium currents-currents that are modulated by CB1 receptors in bipolar cells of the tiger salamander. Presence of CB1 receptors in photoreceptor terminals would therefore be consistent with presynaptic modulation of synaptic transmission, a role seen for cannabinoids in other parts of the brain. Here we report immunohistochemical and electrophysiological evidence for the presence of functional CB1 receptors in rod and cone photoreceptors of the tiger salamander. The cannabinoid receptor agonist WIN 55212-2 enhances calcium currents of rod photoreceptors by 39% but decreases calcium currents of large single cones by 50%. In addition, WIN 55212-2 suppresses potassium currents of rods and large single cones by 44 and 48%, respectively. Thus functional CB1 receptors, present in the terminals of rod and cone photoreceptors, differentially modulate calcium and potassium currents in rods and large single cones. CB1 receptors are therefore well positioned to modulate neurotransmitter release at the first synapse of the visual system. PMID- 12740410 TI - Three-dimensional ocular kinematics during eccentric rotations: evidence for functional rather than mechanical constraints. AB - Previous studies have reported that the translational vestibuloocular reflex (TVOR) follows a three-dimensional (3D) kinematic behavior that is more similar to visually guided eye movements, like pursuit, rather than the rotational VOR (RVOR). Accordingly, TVOR rotation axes tilted with eye position toward an eye fixed reference frame rather than staying relatively fixed in the head like in the RVOR. This difference arises because, contrary to the RVOR where peripheral image stability is functionally important, the TVOR like pursuit and saccades cares to stabilize images on the fovea. During most natural head and body movements, both VORs are simultaneously activated. In the present study, we have investigated in rhesus monkeys the 3D kinematics of the combined VOR during yaw rotation about eccentric axes. The experiments were motivated by and quantitatively compared with the predictions of two distinct hypotheses. According to the first (fixed-rule) hypothesis, an eye-position-dependent torsion is computed downstream of a site for RVOR/TVOR convergence, and the combined VOR axis would tilt through an angle that is proportional to gaze angle and independent of the relative RVOR/TVOR contributions to the total eye movement. This hypothesis would be consistent with the recently postulated mechanical constraints imposed by extraocular muscle pulleys. According to the second (image stabilization) hypothesis, an eye-position-dependent torsion is computed separately for the RVOR and the TVOR components, implying a processing that takes place upstream of a site for RVOR/TVOR convergence. The latter hypothesis is based on the functional requirement that the 3D kinematics of the combined VOR should be governed by the need to keep images stable on the fovea with slip on the peripheral retina being dependent on the different functional goals of the two VORs. In contrast to the fixed-rule hypothesis, the data demonstrated a variable eye-position-dependent torsion for the combined VOR that was different for synergistic versus antagonistic RVOR/TVOR interactions. Furthermore, not only were the eye-velocity tilt slopes of the combined VOR as much as 10 times larger than what would be expected based on extraocular muscle pulley location, but also eye velocity during antagonistic RVOR/TVOR combinations often tilted opposite to gaze. These results are qualitatively and quantitatively consistent with the image-stabilization hypothesis, suggesting that the eye-position-dependent torsion is computed separately for the RVOR and the TVOR and that the 3D kinematics of the combined VOR are dependent on functional rather than mechanical constraints. PMID- 12740411 TI - Space-time maps and two-bar interactions of different classes of direction selective cells in macaque V-1. AB - We used one-dimensional sparse noise stimuli to generate first-order spatiotemporal maps and second-order two-bar interaction maps for 65 simple and 124 complex direction-selective cells in alert macaque V1. Spatial and temporal phase differences between light and dark space-time maps clearly distinguished simple and complex cell populations. Complex cells usually showed similar direction preferences to light and dark bars, but many of the directional simple cells were much more direction selective to one sign of contrast than the reverse. We show that this is predicted by a simple energy model. Some of the direction-selective simple cells showed multiple space-time-slanted subregions, but others (previously described as S1 cells) had space-time maps that looked like just one subregion of an ordinary simple cell. Both simple and complex cells showed directional interactions (nonlinearities) to pairs of flashed bars (a 2 bar apparent-motion stimulus). The space-time slant of the simple cells correlated with the optimum dX/dT (velocity) of the paired-bar interactions. Some complex cells also showed a space-time slant; the direction of the slant usually correlated with the preferred direction of motion, but the degree of slant correlated with the inferred velocity tuning only when measured by a weighted centroid calculation. Principal components analysis of the simple-cell space-time maps yielded one fast temporally biphasic component and a slower temporally monophasic component. We saw no consistent pattern for the spatial phase of the components, unlike previous reports; however, we show that principal components analysis may not distinguish between spatial offsets and phase offsets. PMID- 12740413 TI - Spectral shape sensitivity contributes to the azimuth tuning of neurons in the cat's inferior colliculus. AB - We recorded high-best-frequency single-unit responses to free-field noise bursts that varied in intensity and azimuth to determine whether inferior colliculus (IC) neurons derive directionality from monaural spectral-shape. Sixty-nine percent of the sample was directional (much more responsive at some azimuths than others). One hundred twenty-nine directional units were recorded under monaural conditions (unilateral ear plugging). Binaural directional (BD) cells showed weak monaural directionality. Monaural directional (MD) cells showed strong monaural directionality, i.e., were much more responsive at some directions than others. Some MD cells were sensitive to both monaural and binaural directional cues. MD cells were monaurally nondirectional in response to tone bursts that lack direction-dependent variation in spectral shape. MD cells were unresponsive to noise bursts at certain azimuths even at high intensities showing that particular spectral shapes inhibit their responses. Two-tone inhibition was stronger where MD cells were unresponsive to noise stimulation than at directions where they were responsive. According to the side-band inhibition model, MD cells derive monaural directionality by comparing energy in excitatory and inhibitory frequency domains and thus should have stronger inhibitory side-bands than BD cells. MD and BD cells showed differences in breadth of excitatory frequency domains, strength of nonmonotonic level tuning, and responsiveness to tones and noise that were consistent with this prediction. Comparison of these data with previous findings shows that strength of spectral inhibition increases greatly between the level of the cochlear nucleus and the IC, and there is relatively little change in strength of spectral inhibition among the IC, auditory thalamus, and cortex. PMID- 12740412 TI - Substructure of direction-selective receptive fields in macaque V1. AB - We used two-dimensional (2-D) sparse noise to map simultaneous and sequential two spot interactions in simple and complex direction-selective cells in macaque V1. Sequential-interaction maps for both simple and complex cells showed preferred direction facilitation and null-direction suppression for same-contrast stimulus sequences and the reverse for inverting-contrast sequences, although the magnitudes of the interactions were weaker for the simple cells. Contrast-sign selectivity in complex cells indicates that direction-selective interactions in these cells must occur in antecedent simple cells or in simple-cell-like dendritic compartments. Our maps suggest that direction selectivity, and on and off segregation perpendicular to the orientation axis, can occur prior to receptive-field elongation along the orientation axis. 2-D interaction maps for some complex cells showed elongated alternating facilitatory and suppressive interactions as predicted if their inputs were orientation-selective simple cells. The negative interactions, however, were less elongated than the positive interactions, and there was an inflection at the origin in the positive interactions, so the interactions were chevron-shaped rather than band-like. Other complex cells showed only two round interaction regions, one negative and one positive. Several explanations for the map shapes are considered, including the possibility that directional interactions are generated directly from unoriented inputs. PMID- 12740414 TI - Spindle model responsive to mixed fusimotor inputs and testable predictions of beta feedback effects. AB - Skeletofusimotor (beta) motoneurons innervate both extrafusal muscle units and muscle fibers within muscle spindle stretch receptors. By receiving excitation from group Ia muscle spindle afferents and driving the muscle spindle afferents that excite them, they form a positive feedback loop of unknown function. To study it, we developed a computationally efficient model of group Ia afferent behavior, capable of responding to multiple fusimotor inputs, that matched experimental data. This spindle model was then incorporated into a simulation of group Ia feedback during ramp/hold and triangular stretches with and without closure of the beta loop, assuming that gamma and beta fusimotor drives of the same type (static or dynamic) have identical effects on spindle afferent firing. The effects of beta feedback were implemented by driving a fusimotor input with a delayed and filtered fraction of the spindle afferent output. During triangular stretches, feedback through static beta motoneurons enhanced Ia afferent firing during shortening of the spindle. In contrast, closure of a dynamic beta loop increased Ia firing during lengthening. The strength of beta feedback, estimated as a "loop gain" was comparable to experimental estimates. The loop gain increased with velocity and amplitude of stretch but decreased with increased superimposed gamma fusimotor rates. The strongest loop gains were seen when the beta loop and the gamma bias were of different types (static vs. dynamic). PMID- 12740415 TI - Three-dimensional eye-head coordination is implemented downstream from the superior colliculus. AB - How the brain transforms two-dimensional visual signals into multi-dimensional motor commands, and subsequently how it constrains the redundant degrees of freedom, are fundamental problems in sensorimotor control. During fixations between gaze shifts, the redundant torsional degree of freedom is determined by various neural constraints. For example, the eye- and head-in-space are constrained by Donders' law, whereas the eye-in-head obeys Listing's law. However, where and how the brain implements these laws is not yet known. In this study, we show that eye and head movements, elicited by unilateral microstimulations of the superior colliculus (SC) in head-free monkeys, obey the same Donders' strategies observed in normal behavior (i.e., Listing's law for final eye positions and the Fick strategy for the head). Moreover, these evoked movements showed a pattern of three-dimensional eye-head coordination, consistent with normal behavior, where the eye is driven purposely out of Listing's plane during the saccade portion of the gaze shift in opposition to a subsequent torsional vestibuloocular reflex slow phase, such that the final net torsion at the end of each head-free gaze shift is zero. The required amount of saccade related torsion was highly variable, depending on the initial position of the eye and head prior to a gaze shift and the size of the gaze shift, pointing to a neural basis of torsional control. Because these variable, context-appropriate torsional saccades were correctly elicited by fixed SC commands during head-free stimulations, this shows that the SC only encodes the horizontal and vertical components of gaze, leaving the complexity of torsional organization to downstream control systems. Thus we conclude that Listing's and Donders' laws of the eyes and head, and their three-dimensional coordination mechanisms, must be implemented after the SC. PMID- 12740416 TI - Morphological and physiological characterization of layer VI corticofugal neurons of mouse primary visual cortex. AB - Layer VI is the origin of the massive feedback connection from the cortex to the thalamus, yet its complement of cell types and their connections is poorly understood. The physiological and morphological properties of corticofugal neurons of layer VI of mouse primary visual cortex were investigated in slices loaded with the Ca(2+) indicator fura-2AM. To identify corticofugal neurons, electrical stimulation of the white matter (WM) was done in conjunction with calcium imaging to detect neurons that responded with changes in intracellular Ca(2+) concentrations in response to the stimulation. Subsequent whole cell recordings confirmed that they discharged antidromic action potentials after WM stimulation. Antidromically activated neurons were more excitable and had different spiking properties than neighboring nonantidromic neurons, although both groups had similar input resistances. Furthermore, antidromic neurons possessed narrower action potentials and smaller afterhyperpolarizations. Additionally, three-dimensional reconstructions indicated that antidromically activated neurons had a distinct morphology with longer apical dendrites and fewer nonprimary dendrites than nonantidromic cells. To identify the antidromic neurons, rhodamine microspheres were injected into the dorsal lateral geniculate nucleus of the thalamus and allowed to retrogradely transport back to the somata of the layer VI cortico-geniculate neurons. Physiological and anatomical analysis indicated that most antidromic neurons were likely to be cortico-geniculate neurons. Our results show that cortico-thalamic neurons represent a specific functional and morphological class of layer VI neurons. PMID- 12740417 TI - Representation of the temporal order of visual objects in the primate lateral prefrontal cortex. AB - Recollecting a past episode involves remembering the temporal order of events. We studied cellular activity in the lateral prefrontal cortex (PFC) of two monkeys that were required to remember the temporal order in which visual objects appeared and to reach for each object in the same order after a delay. Here, we report the cellular activity in the lateral PFC, focusing on the delay period. We found that 43% of the delay-period activity was selective for the sequence in which the visual objects were presented during the cue period. While the majority of cellular activity was selective for multiple sequences, some cells (31%) were selective for only one of six sequences. Our findings show that PFC cells are involved in holding temporal order information when that information is necessary for planning forthcoming motor behavior. PMID- 12740418 TI - Integrins regulate NMDA receptor-mediated synaptic currents. AB - Synapses contain high concentrations of integrins, adhesion receptors known to influence the operation of neighboring transmembrane proteins. Evidence that integrins are important for consolidation of long-term potentiation suggests that these adhesion proteins may modulate activities of synaptic glutamate receptors. The present study provides a first test of the possibility that integrins modulate synaptic N-methyl-d-aspartate (NMDA)-type glutamate receptor activities. Excitatory postsynaptic currents (EPSCs) were recorded with whole cell clamp from hippocampal slices in which AMPA-type glutamate receptors and GABA(A) receptors were pharmacologically blocked. Microperfusion of the peptide integrin ligand gly arg-gly-asp-ser-pro (GRGDSP) caused an approximately twofold increase in the amplitude and duration of NMDA receptor-gated synaptic currents. Control peptides had no effect. Paired-pulse facilitation was unchanged, indicating that the ligand did not modify neurotransmitter release probabilities. Infusion of the Src kinase antagonist PP2 but not the control drug 4-amino-7-phenylpyrazolo[3,4 d]pyrimidine eliminated the enhancing effect of GRGDSP. Integrins regulate Src kinases that are known to phosphorylate NMDA receptors. It is concluded that integrins act through this route to exert potent modulatory effects on the operation of NMDA receptors. PMID- 12740419 TI - Role of the dorsolateral pontine nucleus in short-term adaptation of the horizontal vestibuloocular reflex. AB - The dorsolateral pontine nucleus (DLPN) is a major component of the cortico-ponto cerebellar pathway that carries signals essential for smooth pursuit. This pathway also carries visual signals that could play a role in visually guided motor learning in the vestibular ocular reflex (VOR). However, there have been no previous studies that tested this possibility directly. The aim of this study was to determine the potential role of the DLPN in short-term VOR gain adaptation produced by viewing a scene through lenses placed in front of both eyes. In control experiments, adaptation of VOR gain was achieved by sinusoidal rotation (0.2 Hz, 30 degrees /s) for 2 h while the monkey viewed a stationary visual surround through either magnifying (x2) or minifying (x0.5) lenses. This led to increases (23-32%) or decreases (22-48%) of VOR gain as measured in complete darkness (VORd). We used injections of muscimol, a potent GABA(A) agonist (0.5 microl; 2%), to reversibly inactivate the DLPN, unilaterally, in three monkeys. After DLPN inactivation, initial acceleration of ipsilateral smooth-pursuit was reduced by 35-68%, and steady-state gain was reduced by 32-61%. Despite these significant deficits (P < 0.01) in ipsilesional smooth pursuit, the VOR during lens viewing was similar to that measured in preinjection control experiments. Similarly, after 2 h of adaptation, VORd gain was not significantly different (P > 0.61) from control adaptation values for either ipsi- or contralesional directions of head rotation. This was the case even though a stable ipsilesional smooth pursuit deficit persisted throughout the full adaptation period. Our results suggest that visual error signals for short-term adaptation of the VOR are derived from sources other than the DLPN perhaps including other basilar pontine nuclei and the accessory optic system. PMID- 12740420 TI - An important role for the Na+-Ca2+ exchanger in the decrease in cytosolic Ca2+ concentration induced by isoprenaline in the porcine coronary artery. AB - The role of the Na+-Ca2+ exchanger (NCX) in the mechanism of the isoprenaline (Iso)-induced vasorelaxation was investigated by simultaneously monitoring the intracellular Ca2+ concentration ([Ca2+]i) and tension of fura-2-loaded medial strips of porcine coronary arteries. Normal physiological salt solution (PSS) contained 137.3 mM Na+ and 5.9 mM K+. During the sustained phase of contraction, Iso induced only a transient decrease in [Ca2+]i when contraction was induced by depolarization with 118 mM K+ solution containing 25.2 mM Na+. When contraction was induced with 30 mM K+ in PSS containing 113.2 mM Na+, Iso induced a sustained decrease in [Ca2+]i, whereas in contractions induced by 30 mM K+ in a low Na+ (25.2 mM Na+) PSS, Iso transiently decreased [Ca2+]i. Replacement of Ca2+ with Ba2+ (which cannot be extruded by the Ca2+ pumps but can be extruded through the NCX) resulted in decreased [Ba2+]i induced by Iso in normal but not in low Na+ PSS. On the other hand, Iso induced a sustained decrease in [Ca2+]i when strips were pre-contracted by U46619, a thromboxane A2 analogue, in PSS. Various types of K+ channel blockers (iberiotoxin, 4-aminopyridine, apamin or glibenclamide) or combinations of these blockers failed to completely inhibit the Iso-induced decreases in [Ca2+]i and tension. However, Iso-induced sustained decreases in [Ca2+]i during the contraction induced by U46619 were greatly inhibited in a low Na+ PSS. The Iso-induced decrease in tension during contraction by U46619 was greatly inhibited by 2',4'-dichlorobenzamil, a forward- and reverse-mode NCX inhibitor, but not by ouabain, a selective inhibitor of Na+,K+-ATPase. These results indicate that the NCX is involved in the Iso-induced reduction of [Ca2+]i and tension of the porcine coronary arterial smooth muscle. PMID- 12740421 TI - Biophysical and pharmacological characterization of voltage-gated calcium currents in turtle auditory hair cells. AB - Hair cell calcium channels regulate membrane excitability and control synaptic transmission. The present investigations focused on determining whether calcium channels vary between hair cells of different characteristic frequencies or if multiple channel types exist within a hair cell, each serving a different function. To this end, turtle auditory hair cells from high- (317 +/- 27 Hz) and low-frequency (115 +/- 6 Hz) positions were voltage clamped using the whole-cell recording technique, and calcium currents were characterized based on activation, inactivation and pharmacological properties. Pharmacological sensitivity to dihydropyridines (nimodipine, Bay K 8644), benzothiazepines (diltiazem) and acetonitrile derivatives (verapamil, D600) and the insensitivity to non-L-type calcium channel antagonists support the conclusion that only L-type calcium channels were present. Fast activation rise times (< 0.5 ms), hyperpolarized half activation potentials and a relative insensitivity to nimodipine suggest the channels were of the alpha1D (CaV1.3) variety. Although no pharmacological differences were found between calcium currents obtained from high- and low frequency cells, low-frequency cells activated slightly faster and at hyperpolarized potentials, with half-activating voltages of -43 +/- 1 mV compared to -35 +/- 1 mV. Inactivation was observed in both high- and low-frequency cells. The time course of inactivation required three time constants for a fit. Long depolarizations could result in complete inactivation. The voltage of half inactivation was -40 +/- 2 mV for high-frequency cells and -46 +/- 2 mV for low frequency cells. Calcium channel inactivation did not significantly alter hair cell electrical resonant properties elicited from protocols where the membrane potential was hyperpolarized or depolarized prior to characterizing the resonance. A bell-shaped voltage dependence and modest sensitivities to intracellular calcium chelators and external barium ions suggest that inactivation was calcium dependent. PMID- 12740422 TI - Evolution of learning in three aplysiid species: differences in heterosynaptic plasticity contrast with conservation in serotonergic pathways. AB - We investigated the neurobiological basis of variation in sensitization between three aplysiid species: Aplysia californica, Phyllaplysia taylori and Dolabrifera dolabrifera. We tested two different forms of sensitization induced by a noxious tail shock: local sensitization, expressed near the site of shock, and general sensitization, tested at remote sites. Aplysia showed both local and general sensitization, whereas Phyllaplysia demonstrated only local sensitization, and Dolabrifera lacked both forms of learning. We then investigated a neurobiological correlate of sensitization, heterosynaptic modulation of sensory neuron excitability by tail-nerve stimulation. We found (1) an increase in sensory neuron (SN) excitability after both ipsilateral and contralateral nerve stimulation in Aplysia, (2) a smaller and shorter-lasting increase in Phyllaplysia, and (3) no effect in Dolabrifera. Because sensitization in Aplysia is strongly correlated with serotonergic (5-HT) neuromodulation, we hypothesized that the observed interspecific variation in sensitization and SN neuromodulation might be correlated with variation in the anatomy and/or functional response of the serotonergic system. However, using immunohistochemistry, we found that all three species showed a similar pattern of 5-HT innervation. Furthermore, they also showed comparable 5-HT release evoked by tail-nerve shock, as measured with chronoamperometry. These observations indicate that interspecific variation in learning is correlated with differences in SN heterosynaptic plasticity within a background of evolutionary conservation in the 5-HT neuromodulatory pathway. We thus hypothesize that evolutionary changes in learning phenotype do not involve modifications of the 5-HT pathway per se, but rather, changes in the response of SNs to the activation of this or other neuromodulatory pathways upon noxious stimulation. PMID- 12740424 TI - Manipulation of CD98 expression affects both trophoblast cell fusion and amino acid transport activity during syncytialization of human placental BeWo cells. AB - The physiological importance of CD98 surface antigen in regulating placental trophoblast cell fusion and amino acid transport activity has been studied in parallel in a cell model of syncytialization (the cytotrophoblast cell line BeWo following increased intracellular cAMP by forskolin treatment) using antisense oligonucleotides. CD98 protein abundance (determined by Western blot) was decreased by 50 % following antisense oligonucleotide transfection. Transfection with antisense oligonucleotide altered the responses of BeWo to forskolin. Cell fusion (determined by a quantitative flow cytometry assay) was inhibited by 57 %, and both human chorionic gonadotropin secretion and L-leucine influx through system L were suppressed. These findings show that CD98 is involved in the process of cell fusion necessary for syncytiotrophoblast formation and that during this physiologically important event, amino acid transport activity is also regulated through expression of this membrane protein. PMID- 12740423 TI - The relationship between transplacental O2 diffusion and placental expression of PlGF, VEGF and their receptors in a placental insufficiency model of fetal growth restriction. AB - Placental growth factor (PlGF) and vascular endothelial growth factor (VEGF) are involved in placental angiogenesis through interactions with the VEGFR-1 and VEGFR-2 receptors. The placenta of pregnancies whose outcome is fetal growth restriction (FGR) are characterized by abnormal angiogenic development, classically associated with hypoxia. The present study evaluated the near-term expression of this growth factor family in an ovine model of placental insufficiency-FGR, in relationship to uteroplacental oxygenation. Compared to controls, FGR pregnancies demonstrated a 37% increase in uterine blood flow (FGR vs. control, 610.86+/-48.48 vs. 443.17+/-37.39 ml min(-1) (kg fetus)(-1); P<0.04), which was associated with an increased maternal uterine venous PO2 (58.13+/-1.00 vs. 52.89+/-1.26 mmHg; P<0.02), increased umbilical artery systolic/diastolic ratio (3.90+/-0.33 vs. 2.12+/-0.26, P<0.05), and fetal hypoxia (arterial PO2; 12.79+/-0.97 vs. 18.65+/-1.6 mmHg, P<0.005). Maternal caruncle PlGF mRNA was increased in FGR (P<0.02), while fetal cotyledon VEGF mRNA was reduced (P<0.02). VEGFR-1 mRNA was also reduced in FGR fetal cotyledon (P<0.001) but was not altered in caruncle tissue. Immunoblot analysis of PlGF and VEGF demonstrated single bands at 19,000 and 18,600 Mr, respectively. Caruncle PlGF concentration was increased (P<0.04), while cotyledon VEGF was decreased (P<0.05) in FGR placentae. The data establish that uterine blood flow is not reduced in relationship to metabolic demands in this FGR model and that the transplacental PO2 gradient is increased, maintaining umbilical oxygen uptake per unit of tissue. Furthermore, these data suggest that an increased transplacental gradient of oxygen generates changes in angiogenic growth factors, which may underline the pathophysiology of the post-placental hypoxic FGR. PMID- 12740426 TI - Insulin signalling and resistance in patients with chronic heart failure. AB - We investigated whether insulin resistance in patients with chronic heart failure (CHF) is associated with impaired insulin signalling in skeletal muscle and whether exercise training would lead to an improvement in insulin signalling, concomitant with enhanced insulin action. Fourteen men with CHF due to idiopathic dilated cardiomyopathy, with mild-to-moderate limitation of physical activity and a left-ventricular ejection fraction of less than 45 %, were studied before and after either a 5 month exercise training programme (n = 7) or standard care (n = 7). Seven healthy men participated as controls. Whole-body insulin-stimulated glucose uptake was determined by the euglycaemic hyperinsulinaemic clamp technique and skeletal muscle biopsy samples were obtained before and after the insulin infusion for insulin signalling measurements. Insulin-stimulated glucose uptake was 20 % lower in CHF patients versus healthy subjects. Physiological hyperinsulinaemia increased tyrosine phosphorylation of insulin receptor substrate (IRS)-1 by approximately 2.5-fold, IRS-1-associated phosphatidylinositol 3-kinase (PI-3-kinase) activity by approximately 2-fold and Akt (protein kinase B) phosphorylation by approximately 3-fold, with similar responses between healthy subjects and CHF patients. Insulin-mediated glucose uptake was not altered in patients after standard care, whereas exercise training elicited a 25 % increase in glucose uptake. Neither standard care nor exercise training altered insulin-stimulated tyrosine phosphorylation of IRS-1, IRS-1 associated PI-3-kinase activity or Akt phosphorylation. In conclusion, the CHF patients demonstrated impaired insulin-stimulated glucose uptake, despite normal signal transduction in skeletal muscle at the level of IRS-1, PI-3-kinase and Akt. Of clinical relevance is the finding that exercise training improves glucose uptake. However, these changes in insulin action after exercise training appear to be independent of enhanced insulin signalling at the level of IRS-1, PI-3 kinase or Akt. PMID- 12740427 TI - Inward rectification by polyamines in mouse Kir2.1 channels: synergy between blocking components. AB - We recently characterized two distinct mechanisms by which the polyamine spermine blocks Kir2.1 channels: (1) by reduction of negative surface charges in the cytoplasmic pore, thereby reducing single-channel conductance, and (2) by direct open channel transmembrane pore block. The extent to which the surface charge reduction component is mediated by passive surface charge screening versus binding of polyamines to these charges, as well as the extent to which the surface charge reduction and pore block mechanisms are synergistic, versus simply additive, was not established. To address these issues, macroscopic currents were recorded from inside-out giant patches from Xenopus oocytes and from single channel currents from COS7 cells expressing wild-type and mutant Kir2.1 channels, during exposure to polyamines of varying length and charge. The surface charge reduction component was decreased when polyamine charge (at constant length) was decreased from 4 (spermine) to 2 (diamine 10, DA10). Moreover, the surface charge reduction component of block involved more than passive surface charge screening and required binding of polyamines to the cytoplasmic pore, since it was eliminated when polyamine length was shortened below six alkyl groups. Loss of surface charge reduction also dramatically affected open channel pore block. The latter consisted of two subcomponents with fast and slow kinetics, respectively. The slow subcomponent decreased as blocker length decreased (DA10, DA8 and DA6), whereas the fast subcomponent was sensitive to blocker charge (spermine vs. DA10). Neutralization of E224 and E299, which eliminated the surface charge reduction component of block, also eliminated the fast subcomponent of pore block. Neutralization of D172 had no effect on the surface charge reduction component, but weakened both of the subcomponents of pore block. These findings can be accounted for by a model in which the negative charges at E224, E299 and D172 act in a concerted manner to coordinate the surface charge reduction and open channel components of polyamine block. In this model, the binding of polyamines to surface charges E224 and E299 pre-positions them in the cytoplasmic pore in a manner that directly facilitates their entry and exit from a transmembrane pore-occluding site involving D172. A molecular model using the recently reported 1.8 A resolution structure of the inward-rectifier cytoplasmic pore, adapted to Kir2.1, is consistent with longer polyamines binding at their positively charged ends to the E224 and E299 positions in the same subunit, potentially accommodating four polyamine molecules per channel. PMID- 12740429 TI - Purinergic and adrenergic Ca2+ transients during neurogenic contractions of rat mesenteric small arteries. AB - Contraction of small arteries is regulated by the sympathetic nervous system, but the Ca2+ transients during neurally stimulated contraction of intact small arteries have not yet been recorded. We loaded rat mesenteric small arteries with the fluorescent Ca2+ indicator fluo-4 and mounted them in a myograph that permitted simultaneous (i) high-speed confocal imaging of fluorescence from individual smooth muscle cells, (ii) electrical stimulation of perivascular nerves, and (iii) recording of isometric tension. Sympathetic neuromuscular transmission was achieved by electrical field stimulation (EFS) (frequency, 10 Hz; pulse voltage, 40 V; pulse duration, 0.2 ms) in the presence of capsaicin and scopolamine (to inhibit 'sensory' and cholinergic nerves, respectively). During the first 20 s of EFS, force rose to a small peak and then declined. During this time, junctional Ca2+ transients (jCaTs) were present at relatively high frequency. We have previously attributed jCaTs to influx of Ca2+ through post junctional P2X receptors activated by ATP. Propagating asynchronous Ca2+ waves, previously associated with bath-applied alpha1-adrenoceptor agonists, were not initially present. During the next 2.5 min of EFS, force rose slowly, and asynchronous propagating Ca2+ waves appeared. The selective alpha1-adrenoceptor antagonist prazosin abolished both the slowly developing contraction and the Ca2+ waves, but reduced the initial transient contraction by only ~25 %. During 3 min of EFS in prazosin, the frequency of jCaTs declined markedly; at sites at which at least one jCaT occurred, the average probability of a jCaT was 0.008 +/- 0.002 pulse-1 in the first 20 s and 0.0007 +/- 0.0002 pulse-1 in the last 20 s. We suggest that (i) ATP released from sympathetic varicosities activates the initial, transient, contraction and the activator Ca2+ is derived largely from jCaTs, and (ii) sympathetically released noradrenaline (NA) activates the later, major contraction through mechanisms involving alpha1-adrenoceptors and which are associated with propagating Ca2+ waves. PMID- 12740430 TI - Inhibition of HERG K+ current and prolongation of the guinea-pig ventricular action potential by 4-aminopyridine. AB - 4-Aminopyridine (4-AP) has been used extensively to study transient outward K+ current (ITO,1) in cardiac cells and tissues. We report here inhibition by 4-AP of HERG (the human ether-a-go-go-related gene) K+ channels expressed in a mammalian cell line, at concentrations relevant to those used to study ITO,1. Under voltage clamp, whole cell HERG current (IHERG) tails following commands to +30 mV were blocked with an IC50 of 4.4 +/- 0.5 mM. Development of block was contingent upon HERG channel gating, with a preference for activated over inactivated channels. Treatment with 5 mM 4-AP inhibited peak IHERG during an applied action potential clamp waveform by ~59 %. It also significantly prolonged action potentials and inhibited resurgent IK tails from guinea-pig isolated ventricular myocytes, which lack an ITO,1. We conclude that by blocking the alpha subunit of the IKr channel, millimolar concentrations of 4-AP can modulate ventricular repolarisation independently of any action on ITO,1. PMID- 12740431 TI - Identification of receptors for pig endogenous retrovirus. AB - Xenotransplantation of porcine tissues has the potential to treat a wide variety of major health problems including organ failure and diabetes. Balanced against the potential benefits of xenotransplantation, however, is the risk of human infection with a porcine microorganism. In particular, the transmission of porcine endogenous retrovirus (PERV) is a major concern [Chapman, L. E. & Bloom, E. T. (2001) J. Am. Med. Assoc. 285, 2304-2306]. Here we report the identification of two, sequence-related, human proteins that act as receptors for PERV-A, encoded by genes located on chromosomes 8 and 17. We also describe homologs from baboon and porcine cells that also are active as receptors. Conversely, activity could not be demonstrated with a syntenic murine receptor homolog. Sequence analysis indicates that PERV-A receptors [human PERV-A receptor (HuPAR)-1, HuPAR-2, baboon PERV-A receptor 2, and porcine PERV-A receptor] are multiple membrane-spanning proteins similar to receptors for other gammaretroviruses. Expression is widespread in human tissues including peripheral blood mononuclear cells, but their biological functions are unknown. The identification of the PERV-A receptors opens avenues of research necessary for a more complete assessment of the retroviral risks of pig to human xenotransplantation. PMID- 12740432 TI - Development of teeth in chick embryos after mouse neural crest transplantations. AB - Teeth were lost in birds 70-80 million years ago. Current thinking holds that it is the avian cranial neural crest-derived mesenchyme that has lost odontogenic capacity, whereas the oral epithelium retains the signaling properties required to induce odontogenesis. To investigate the odontogenic capacity of ectomesenchyme, we have used neural tube transplantations from mice to chick embryos to replace the chick neural crest cell populations with mouse neural crest cells. The mouse/chick chimeras obtained show evidence of tooth formation showing that avian oral epithelium is able to induce a nonavian developmental program in mouse neural crest-derived mesenchymal cells. PMID- 12740433 TI - Liquid-vapor oscillations of water in hydrophobic nanopores. AB - Water plays a key role in biological membrane transport. In ion channels and water-conducting pores (aquaporins), one-dimensional confinement in conjunction with strong surface effects changes the physical behavior of water. In molecular dynamics simulations of water in short (0.8 nm) hydrophobic pores the water density in the pore fluctuates on a nanosecond time scale. In long simulations (460 ns in total) at pore radii ranging from 0.35 to 1.0 nm we quantify the kinetics of oscillations between a liquid-filled and a vapor-filled pore. This behavior can be explained as capillary evaporation alternating with capillary condensation, driven by pressure fluctuations in the water outside the pore. The free-energy difference between the two states depends linearly on the radius. The free-energy landscape shows how a metastable liquid state gradually develops with increasing radius. For radii > approximately 0.55 nm it becomes the globally stable state and the vapor state vanishes. One-dimensional confinement affects the dynamic behavior of the water molecules and increases the self diffusion by a factor of 2-3 compared with bulk water. Permeabilities for the narrow pores are of the same order of magnitude as for biological water pores. Water flow is not continuous but occurs in bursts. Our results suggest that simulations aimed at collective phenomena such as hydrophobic effects may require simulation times >50 ns. For water in confined geometries, it is not possible to extrapolate from bulk or short time behavior to longer time scales. PMID- 12740434 TI - T helper type 1 lymphocytes drive inflammation in human atherosclerotic lesions. AB - Atherosclerotic lesions are infiltrated by macrophages and T lymphocytes, potentially reactive to pathogens. We studied in vivo activated T lymphocytes that infiltrate atherosclerotic plaques of Helicobacter pylori-infected patients with or without anti-Chlamydia pneumoniae antibodies. In all atherosclerotic lesions, T helper type 1 (Th1) cells were predominant. C. pneumoniae-specific T cells were detected only in the plaques of anti-C. pneumoniae seropositive patients, whereas H. pylori-specific T cells were found in the gastric mucosa but not in the plaques of the same patients. Plaque-derived Th1 cells expressed cytotoxicity, proapoptotic activity, and help for monocyte tissue factor production. Although multifactorial, atherosclerosis can be regarded as a Th1 driven immunopathological condition. PMID- 12740435 TI - Purine synthesis and increased Agrobacterium tumefaciens transformation of yeast and plants. AB - The bacterium Agrobacterium tumefaciens transforms eukaryotic hosts by transferring DNA to the recipient cell where it is integrated and expressed. Bacterial factors involved in this interkingdom gene transfer have been described, but less is known about host-cell factors. Using the yeast Saccharomyces cerevisiae as a model host, we devised a genetic screen to identify yeast mutants with altered transformation sensitivities. Twenty-four adenine auxotrophs were identified that exhibited supersensitivity to A. tumefaciens mediated transformation when deprived of adenine. We extended these results to plants by showing that purine synthesis inhibitors cause supersensitivity to A. tumefaciens transformation in three plant species. The magnitude of this effect is large and does not depend on prior genetic manipulations of host cells. These data indicate the utility of yeast as a model for the transformation process and identify purine biosynthesis as a key determinant of transformation efficiency. These findings should increase the utility of A. tumefaciens in genetic engineering. PMID- 12740436 TI - Understanding tubulin-Taxol interactions: mutations that impart Taxol binding to yeast tubulin. AB - We have successfully used mutagenesis to engineer Taxol (paclitaxel) binding activity in Saccharomyces cerevisiae tubulin. Taxol, a successful antitumor agent, acts by promoting tubulin assembly and stabilizing microtubules. Several structurally diverse antimitotic compounds, including the epothilones, compete with Taxol for binding to mammalian microtubules, suggesting that Taxol and these compounds share an overlapping binding site. However, Taxol has no effect on tubulin or microtubules from S. cerevisiae, whereas epothilone does. After considering data on Taxol binding to mammalian tubulin and recent modeling studies, we have hypothesized that differences in five key amino acids are responsible for the lack of Taxol binding to yeast tubulin. After changing these amino acids to those found in mammalian brain tubulin, we observed Taxol-related activity in yeast tubulin comparable to that in mammalian tubulin. Importantly, this experimental system can be used to reveal tubulin interactions with Taxol, the epothilones, and other Taxol-like compounds. PMID- 12740437 TI - A fragment of anthrax lethal factor delivers proteins to the cytosol without requiring protective antigen. AB - Anthrax protective antigen (PA) is a 735-aa polypeptide that facilitates the exit of anthrax lethal factor (LF) from the endosome to the cytosol where the toxin acts. We recently found, however, that a fusion protein of the detoxified N terminal domain of lethal factor (LFn) with a foreign peptide could induce CD8 T cell immune responses in the absence of PA. Because CD8 T cells recognize peptides derived from proteins degraded in the cytosol, this result suggests that lethal factor may be capable of entering the cytosol independently of PA. To investigate this further, the intracellular trafficking of an LFn-enhanced green fluorescent protein fusion protein (LFn-GFP) in the presence or absence of PA was examined by using confocal microscopy. LFn-GFP is able to enter the cytosol without PA. Moreover, it efficiently colocalizes with the proteosome 20s subunit, which degrades proteins into peptides for presentation to CD8 T cells by the MHC class I pathway. We further demonstrate that in the presence of an immune adjuvant LFn fusion protein without PA is able to effectively elicit anti-HIV cytotoxic T lymphocyte in inbred mice. These results indicate that LFn may be used without PA in a protein vaccine as a carrier to deliver antigens into the cytosol for efficient induction of T lymphocyte responses. Furthermore, these results enable us to propose a modified molecular mechanism of anthrax lethal toxin. PMID- 12740438 TI - Mitochondrial succinic-semialdehyde dehydrogenase of the gamma-aminobutyrate shunt is required to restrict levels of reactive oxygen intermediates in plants. AB - The gamma-aminobutyrate (GABA) shunt is a metabolic pathway that bypasses two steps of the tricarboxylic acid cycle, and it is present in both prokaryotes and eukaryotes. In plants the pathway is composed of the calcium/calmodulin-regulated cytosolic enzyme glutamate decarboxylase and the mitochondrial enzymes GABA transaminase and succinic-semialdehyde dehydrogenase (SSADH). The activity of the GABA shunt in plants is rapidly enhanced in response to various biotic and abiotic stresses. However the physiological role of this pathway remains obscure. To elucidate its role in plants, we analyzed Arabidopsis T-DNA knockout mutants of SSADH, the ultimate enzyme of the pathway. Four alleles of the ssadh mutation were isolated, and these exhibited a similar phenotype. When exposed to white light (100 micromol of photons per m2 per s), they appear dwarfed with necrotic lesions. Detailed spectrum analysis revealed that UV-B has the most adverse effect on the mutant phenotype, whereas photosynthetic active range light has a very little effect. The ssadh mutants are also sensitive to heat, as they develop necrosis when submitted to such stress. Moreover, both UV and heat cause a rapid increase in the levels of hydrogen peroxide in the ssadh mutants, which is associated with enhanced cell death. Surprisingly, our study also shows that trichomes are hypersensitive to stresses in ssadh mutants. Our work establishes a role for the GABA shunt in preventing the accumulation of reactive oxygen intermediates and cell death, which appears to be essential for plant defense against environmental stress. PMID- 12740439 TI - Gamma-secretase is a membrane protein complex comprised of presenilin, nicastrin, Aph-1, and Pen-2. AB - gamma-Secretase catalyzes the intramembrane proteolysis of Notch, beta-amyloid precursor protein, and other substrates as part of a new signaling paradigm and as a key step in the pathogenesis of Alzheimer's disease. This unusual protease has eluded identification, though evidence suggests that the presenilin heterodimer comprises the catalytic site and that a highly glycosylated form of nicastrin associates with it. The formation of presenilin heterodimers from the holoprotein is tightly gated by unknown limiting cellular factors. Here we show that Aph-1 and Pen-2, two recently identified membrane proteins genetically linked to gamma-secretase, associate directly with presenilin and nicastrin in the active protease complex. Coexpression of all four proteins leads to marked increases in presenilin heterodimers, full glycosylation of nicastrin, and enhanced gamma-secretase activity. These findings suggest that the four membrane proteins comprise the limiting components of gamma-secretase and coassemble to form the active enzyme in mammalian cells. PMID- 12740441 TI - Control of seed germination in transgenic plants based on the segregation of a two-component genetic system. AB - We have developed a repressible seed-lethal (SL) system aimed at reducing the probability of transgene introgression into a population of sexually compatible plants. To evaluate the potential of this method, tobacco plants were transformed with an SL construct comprising gene 1 and gene 2 from Agrobacterium tumefaciens whereby gene 1 was controlled by the seed-specific phaseolin promoter modified to contain a binding site for the Escherichia coli TET repressor (R). The expression of this construct allows normal plant and seed development but inhibits seed germination. Plants containing the SL construct were crossed with plants containing the tet R gene to derive plant lines where the expression of the SL construct is repressed. Plant lines that contained both constructs allowed normal seed formation and germination, whereas seeds in which the SL construct was separated from the R gene through segregation did not germinate. The requirements of such a method to efficiently control the flow of novel traits among sexually compatible plants are discussed. PMID- 12740440 TI - Structural and biochemical studies of p21Ras S-nitrosylation and nitric oxide mediated guanine nucleotide exchange. AB - Ras is a guanine nucleotide-binding protein that cycles between inactive GDP bound and active GTP-bound states to regulate a diverse array of cellular processes, including cell growth, apoptosis, and differentiation. The guanine nucleotide-bound state of Ras is tightly maintained by regulatory factors to promote regulated growth control. A class of regulatory molecules that lead to Ras activation are guanine nucleotide exchange factors (GEFs). Ras GEFs bind to Ras and facilitate GDP release, followed by GTP incorporation and Ras activation. Nitric oxide (NO) has also been shown to promote guanine nucleotide exchange (GNE) on Ras and increase cellular Ras-GTP levels, but the process by which NO mediated GNE occurs is not clear. We initiated NMR structural and biochemical studies to elucidate how nitrosylation of Ras might lead to enhanced GNE. Surprisingly, our studies show that stable S-nitrosylation of Ras at Cys-118, does not affect the structure of Ras, its association with the Ras-binding domain of Raf (a downstream effector of Ras), or GNE rates relative to non-nitrosylated Ras. We have found, however, that the actual chemical process of nitrosylation, rather than the end-product of Ras S-nitrosylation, accounts for the enhanced GNE that we have observed and that has been previously observed by others. PMID- 12740442 TI - Sympatric speciation through intraspecific social parasitism. AB - Sympatric speciation through intraspecific social parasitism has been proposed for the evolution of Hymenopteran workerless parasites. Such inquilines exploit related host taxa to produce their own sexual offspring. The relatedness of inquilines to their hosts has been generalized in Emery's rule, suggesting that social parasites are close or the closest relatives to their host species. If the closest relative of each parasite is its host, then multiple independent origins of the parasite species are implied even within a single genus, probably through sympatric speciation. To test the plausibility of sympatric speciation in inquilines, we conducted a mitochondrial DNA phylogenetic analysis in three inquiline-host pairs of Myrmica ant species. We show that congeneric inquilines have originated independently several times. We also show that two of the inqulines are more closely related to their hosts than to any other species. Our results suggest sympatric speciation of Myrmica inquilines. Sympatric speciation is probably facilitated by the social biology and ecology of Myrmica, with polygyny as a prerequisite for the evolution of intraspecific parasitism. PMID- 12740443 TI - PPARgamma and PPARdelta negatively regulate specific subsets of lipopolysaccharide and IFN-gamma target genes in macrophages. AB - Natural and synthetic agonists of the peroxisome proliferator-activated receptor gamma (PPARgamma) regulate adipocyte differentiation, glucose homeostasis, and inflammatory responses. Although effects on adipogenesis and glucose metabolism are genetically linked to PPARgamma, the PPARgamma dependence of antiinflammatory responses of these substances is less clear. Here, we have used a combination of mRNA expression profiling and conditional disruption of the PPARgamma gene in mice to characterize programs of transcriptional activation and repression by PPARgamma agonists in elicited peritoneal macrophages. Natural and synthetic PPARgamma agonists, including the thiazolidinedione rosiglitazone (Ro), modestly induced the expression of a surprisingly small number of genes, several of which were also induced by a specific PPARdelta agonist. The majority of these genes encode proteins involved in lipid homeostasis. In contrast, Ro inhibited induction of broad subsets of lipopolysaccharide and IFN-gamma target genes in a gene-specific and PPARgamma-dependent manner. At high concentrations, Ro inhibited induction of lipopolysaccharide target genes in PPARgamma-deficient macrophages, at least in part by activating PPARdelta. These studies establish overlapping transactivation and transrepression functions of PPARgamma and PPARdelta in macrophages and suggest that a major transcriptional role of PPARgamma is negative regulation of specific subsets of genes that are activated by T helper 1 cytokines and pathogenic molecules that signal through pattern recognition receptors. These findings support a physiological role of PPARgamma in regulating both native and acquired immune responses. PMID- 12740444 TI - Susceptibility to tuberculosis: a locus on mouse chromosome 19 (Trl-4) regulates Mycobacterium tuberculosis replication in the lungs. AB - The mouse DBA/2 (D2) strain is very susceptible to infection with virulent Mycobacterium tuberculosis, whereas C57BL/6 (B6) is much more resistant. Infection of D2 and B6 mice with M. tuberculosis H37Rv by the respiratory route is biphasic: during the first 3 weeks, there is rapid bacterial growth in the lung of both strains, whereas beyond this point replication stops in B6 but continues in D2, causing rapidly fatal pulmonary disease. To identify the genes regulating growth of M. tuberculosis in the lungs of these two strains, 98 informative (B6 x D2) F2 mice were infected by the respiratory route with M. tuberculosis H37Rv (2 x 102 colony-forming units), and the extent of bacterial replication in the lungs at 90 days was used as a quantitative measure of susceptibility in a whole-genome scan. Quantitative trait locus mapping identified a major locus on chromosome 19 (Tuberculosis resistance locus-4, Trl 4; logarithm of odds 5.6), which regulated pulmonary replication of M. tuberculosis and accounted for 25% of the phenotypic variance. B6 alleles at Trl 4 were inherited in an incompletely dominant fashion and associated with reduced bacterial replication. An additional effect of a locus (Trl-3), previously shown to affect survival to i.v. infection with M. tuberculosis, was also noted. F2 mice homozygous for B6 alleles at both Trl-3 and Trl-4 were as resistant as B6 parents, whereas mice homozygous for D2 alleles were as susceptible as D2 parents. These results suggest a strong genetic interaction between Trl-3 and Trl 4 in regulating pulmonary replication of M. tuberculosis. PMID- 12740445 TI - Retrofitting existing chemical scrubbers to biotrickling filters for H2S emission control. AB - Biological treatment is a promising alternative to conventional air-pollution control methods, but thus far biotreatment processes for odor control have always required much larger reactor volumes than chemical scrubbers. We converted an existing full-scale chemical scrubber to a biological trickling filter and showed that effective treatment of hydrogen sulfide (H2S) in the converted scrubber was possible even at gas contact times as low as 1.6 s. That is 8-20 times shorter than previous biotrickling filtration reports and comparable to usual contact times in chemical scrubbers. Significant removal of reduced sulfur compounds, ammonia, and volatile organic compounds present in traces in the air was also observed. Continuous operation for >8 months showed stable performance and robust behavior for H2S treatment, with pollutant-removal performance comparable to that achieved by using a chemical scrubber. Our study demonstrates that biotrickling filters can replace chemical scrubbers and be a safer, more economical technique for odor control. PMID- 12740447 TI - Effects of sibutramine on binge eating, hunger, and fullness in a laboratory human feeding paradigm. AB - OBJECTIVE: The purpose of this study was to evaluate the effects of sibutramine vs. placebo on binge-eating behavior, hunger, and satiety in patients who had problems with binge eating. RESEARCH METHODS AND PROCEDURES: Seven adult subjects who had problems with binge eating (mean age, 42 years) were randomly assigned to receive alternating sibutramine and placebo in a double-blind placebo-controlled crossover study. This involved two 4-week dosing periods separated by a 2-week washout. RESULTS: Subjects lost weight on sibutramine but not on placebo. There was a significant difference in the number of kilocalories consumed between the sibutramine and placebo conditions, with a significant reduction of intake during binge-eating episodes on sibutramine. DISCUSSION: Sibutramine suppresses intake during binge-eating episodes. This effect is demonstrable in a human feeding laboratory paradigm. PMID- 12740446 TI - Disruption of the estrogen receptor beta gene in mice causes myeloproliferative disease resembling chronic myeloid leukemia with lymphoid blast crisis. AB - Proliferation of pluripotent, bone marrow stem cells, which develop to lymphoid and myeloid progenitors, is negatively regulated by estrogen. Although in estrogen deficiency and in estrogen receptor knockout mice there is significant alteration in bone marrow hematopoiesis, the effects of aging on estrogen receptor deficiencies in mice have not been investigated yet. In this study we show that by 1.5 years of age, estrogen receptor beta knockout (ERbeta-/-) mice develop pronounced splenomegaly that is much more severe in females than in males. Further characterization of these mice revealed myelogenous hyperplasia in bone marrow, an increase in the number of granulocytes and B lymphocytes in blood, lymphadenopathy, and infiltration of leukocytes in the liver and lung. Analysis by flow cytometry of the bone marrow cells revealed that the percentage and total number of Gr-1hi/Mac-1hi-positive granulocytes were increased by 15-30% and 100%, respectively. The numbers of B cells in the bone marrow and spleen were significantly higher in ERbeta-/- mice than in WT littermates. Some of the ERbeta /- mice also had a severe lymphoproliferative phenotype. Thus the absence of ERbeta results in a myeloproliferative disease resembling human chronic myeloid leukemia with lymphoid blast crisis. Our results indicate a previously unknown role for ERbeta in regulating the differentiation of pluripotent hematopoietic progenitor cells and suggest that the ERbeta-/- mouse is a potential model for myeloid and lymphoid leukemia. Furthermore, we suggest that ERbeta agonists might have clinical value in the treatment of leukemia. PMID- 12740448 TI - K121Q PC-1 gene polymorphism is not associated with insulin resistance in a Spanish population. AB - OBJECTIVE: To investigate the effect of the K121Q plasma cell membrane glycoprotein (PC-1) polymorphism on the components of the insulin resistance syndrome in a population-based nationwide multicenter study in Spain. RESEARCH METHODS AND PROCEDURES: The subjects of the study were 293 nonrelated adults (44.7% men and 55.3% women) ages 35 to 64 years randomly chosen from a nationwide population-based survey on obesity and related conditions, including insulin resistance and cardiovascular risk factors. Obesity-related anthropometric measurements included blood pressure, oral glucose tolerance test, lipid profile (total cholesterol, high-density lipoprotein- and low-density lipoprotein cholesterol, and triglycerides), plasma leptin, insulin levels by radioimmunoassay, and insulin resistance (homeostasis model assessment). K121Q PC 1 genotypes were determined by restriction fragment-length polymorphism polymerase chain reaction. RESULTS: Overall Q allele frequency was 0.14, with no differences between obese and nonobese individuals (0.15 vs. 0.13). After adjustment for sex, age, BMI, and degree of glucose tolerance, the Q allele was associated with high plasma leptin and triglyceride levels, but not with insulin resistance. DISCUSSION: The results showed that the K121Q PC-1 polymorphism in the Spanish population has no significant impact on insulin sensitivity. PMID- 12740449 TI - Obesity prevention in preschool native-american children: a pilot study using home visiting. AB - OBJECTIVE: To determine whether maternal participation in an obesity prevention plus parenting support (OPPS) intervention would reduce the prevalence of obesity in high-risk Native-American children when compared with a parenting support (PS) only intervention. RESEARCH METHODS AND PROCEDURES: Forty-three mother/child pairs were recruited to participate. Mothers were 26.5 +/- 5 years old with a mean BMI of 29.9 +/- 3 kg/m(2). Children (23 males) were 22 +/- 8 months old with mean weight-for-height z (WHZ) scores of 0.73 +/- 1.4. Mothers were randomly assigned to a 16-week OPPS intervention or PS alone. The intervention was delivered one-on-one in homes by an indigenous peer educator. Baseline and week 16 assessments included weight and height (WHZ score and weight-for-height percentile for children), dietary intake (3-day food records), physical activity (measured by accelerometers), parental feeding style (Child Feeding Questionnaire), and maternal outcome expectations, self-efficacy, and intention to change diet and exercise behaviors. RESULTS: Changes in WHZ scores showed a trend toward significance, with WHZ scores decreasing in the PS condition and increasing among the OPPS group (-0.27 +/- 1.1 vs. 0.31 +/- 1.1, p = 0.06). Children in the OPPS condition also significantly decreased energy intake (-316 +/- 835 kcal/d vs. 197 +/- 608 kcal/d, p < 0.05). Scores on the restriction subscale of the Child Feeding Questionnaire decreased significantly in the OPPS condition (-0.22+/- 0.42 vs. 0.08+/- 0.63, p < 0.05), indicating that mothers in the OPPS group were engaging in less restrictive child feeding practices over time. DISCUSSION: A home-visiting program focused on changing lifestyle behaviors and improving parenting skills showed promise for obesity prevention in high-risk Native-American children. PMID- 12740450 TI - Effects of beta2-adrenergic receptor gene variants on adiposity: the HERITAGE Family Study. AB - We investigated whether the Arg16Gly and Gln27Glu polymorphisms of the beta2 adrenergic receptor gene were associated with body-fat and fat-distribution phenotypes measured before and in response to a 20-week endurance-training program. BMI, fat mass (FAT), percentage of body fat (%FAT), sum of eight skinfolds (SF8), and abdominal fat areas assessed by computed tomography were measured in adult sedentary white and black participants of the HERITAGE Family Study. Evidence of gene-by-obesity interaction was found in whites for several adiposity phenotypes measured before training. Analyses performed separately in nonobese and obese subjects revealed that obese men carrying the Glu27 allele have lower fat accumulation (BMI, FAT, and %FAT) than noncarriers. Among white obese women, Gly16Gly homozygotes had a lower fat accumulation (BMI, FAT, and SF8) than Arg16Gly and Arg16Arg carriers. In response to endurance training, white women with the Arg16Arg genotype exhibited a greater reduction in BMI, FAT, and %FAT. Results observed in blacks were mostly negative. These results suggest that polymorphisms in the beta2-adrenergic receptor gene influence the amount of body fat in white obese men (Gln27Glu) and women (Arg16Gly), as well as the changes in adiposity in response to endurance training in white women (Arg16Gly). PMID- 12740451 TI - Sequence variants of the POMC gene and their associations with body composition in children. AB - We investigated POMC sequence variants in 242 overweight and nonoverweight African-American and white children and examined the associations between body composition and POMC polymorphisms. Three novel polymorphisms and two previously identified sequence variants were found: A7301G, A7429G, and C8246T were all in untranslated regions. A 9-bp (AGC AGC GGC) duplication/insertion was found between positions 7677 and 7678, and one normal-weight African-American girl had a 45-bp triple duplication/insertion at this location. Compared with whites, African-American children were significantly more likely to have polymorphisms A7301G, A7429G, and the 9-bp insertion. However, there were no significant associations between any of the polymorphisms and body composition. Five African American subjects who were homozygous for A7429G had a trend (p = 0.08) for a greater BMI-SD score (5.3 +/- 5.3 kg/m(2)) compared with wild-type children (BMI SD score, 2.4 +/- 3.2 kg/m(2)) or heterozygotes (BMI-SD score, 2.7 +/- 3.7 kg/m(2)). However, there were no differences in BMI-SD score for A7429G when African American subjects were studied separately and both gender and height were taken into account. The contribution of the POMC gene variants we studied to pediatric-onset obesity seems to be limited. PMID- 12740452 TI - Metabolic adaptations to dexamethasone-induced insulin resistance in healthy volunteers. AB - OBJECTIVE: Insulin resistance is observed in individuals with normal glucose tolerance. This indicates that increased insulin secretion can compensate for insulin resistance and that additional defects are involved in impaired glucose tolerance or type 2 diabetes. The objective of this study was to evaluate a procedure aimed at assessing the compensatory mechanisms to insulin resistance. RESEARCH METHODS AND PROCEDURES: Eight healthy nonobese female patients were studied on two occasions, before and after administration of 2 mg/d dexamethasone for 2 days during a two-step hyperglycemic clamp. Insulin secretion was assessed from plasma insulin concentrations. Insulin sensitivity was assessed from the ratio of whole-body glucose use (6,6 (2)H(2) glucose) to plasma insulin concentrations. This procedure is known to induce a reversible impairment of glucose tolerance and insulin resistance. RESULTS: In all subjects, dexamethasone induced a decrease in insulin sensitivity and a proportionate increase in first phase insulin secretion and in insulin concentrations at both steps of glycemia. The resulting hyperinsulinemia allowed the restoration of normal whole-body glucose uptake and the suppression of plasma free fatty acids and triglycerides. In contrast, the suppression of endogenous glucose production was impaired after dexamethasone (p < 0.01). DISCUSSION: Increased insulin secretion fully compensates dexamethasone-induced insulin resistance in skeletal muscle and adipose tissue but not in the liver. This suggests that failure to overcome hepatic insulin resistance can impair glucose tolerance. The compensatory insulin secretion in response to insulin resistance can be assessed by means of a hyperglycemic clamp after a dexamethasone challenge. PMID- 12740453 TI - Decreased uncoupling protein expression and intramyocytic triglyceride depletion in formerly obese subjects. AB - OBJECTIVE: To examine the muscular uncoupling protein expression 2 (UCP2) and UCP3 gene expression in morbid obese subjects before and after bariatric surgery [bilio-pancreatic diversion (BPD)]. RESEARCH METHODS AND PROCEDURES: Eleven obese subjects (BMI = 49 +/- 2 kg/m(2)) were studied before BPD and 24 months after BPD. Skeletal muscle UCP2 and UCP3 mRNA was measured using reverse transcriptase competitive polymerase chain reaction and UCP3 protein by Western blotting. Intramyocytic triglycerides were quantified by high-performance liquid chromatography. Twenty-four-hour energy expenditure and respiratory quotient (RQ) were measured in a respiratory chamber. RESULTS: After BPD, the average weight loss was approximately 38%. Nonprotein RQ was increased in the postobese subjects (0.73 +/- 0.00 vs. 0.83 +/- 0.02, p < 0.001). The intramyocytic triglyceride level dropped (3.66 +/- 0.16 to 1.60 +/- 0.29 mg/100 mg of fresh tissue, p < 0.0001) after BPD. Expression of UCP2 and UCP3 mRNA was significantly reduced (from 35.9 +/- 6.1% to 18.6 +/- 4.5% of cyclophilin, p = 0.02; from 60.2 +/- 14.0% to 33.4 +/- 8.5%, p = 0.03; respectively). UCP3 protein content was also significantly reduced (272.19 +/- 84.13 vs. 175.78 +/- 60.31, AU, p = 0.04). A multiple regression analysis (R(2) = 0.90) showed that IMTG levels (p = 0.007) represented the most powerful independent variable for predicting UCP3 variation. DISCUSSION: The strong correlation of UCP expression and decrease in IMTG levels suggests that triglyceride content plays an even more important role in the regulation of UCP gene expression than the circulating levels of free fatty acids or the achieved degree of weight loss. PMID- 12740454 TI - Assessing dietary and exercise stage of change to optimize weight loss interventions. AB - OBJECTIVE: According to the Transtheoretical Model (TM), tailoring interventions to patients' readiness to change can enhance patient progress and help patients use therapeutic resources more effectively. However, tailoring an intervention depends on accurate assessment of patients' stage of change (SOC). This study describes a multi-item algorithm of SOC (M-SOC) for weight loss-related behaviors that attempts to overcome some of the conceptual and methodological difficulties encountered in previously SOC assessments. RESEARCH METHODS: Data were collected from participants randomly assigned to the treatment arm of a clinical trial comparing a TM-based, cognitive-behavioral intervention with enhanced usual care for weight loss. RESULTS: The stair-step relationships hypothesized by the TM between assessed SOC and key behavioral measures were found for all algorithms (p < 0.05), indicating that the M-SOC algorithm performs as well as single item algorithms. However, 50% fewer patients were classified in action or maintenance for dietary fat intake and portion control (p < 0.05) by the multiple-item algorithms, providing staging more consistent with the clinical presentation of obese individuals. In addition, logistic regression analyses indicated that the single-item algorithms were not sufficient predictors of these behavioral domains. DISCUSSION: To the extent that treatment is guided by assessed SOC, there is potential for a considerable variability in interventions with different approaches to staging. For weight-loss interventions that target portion control and decreased fat intake, the multi-item SOC algorithms seem to be better guides for matching treatments to SOC. PMID- 12740455 TI - Sympathovagal balance, nighttime blood pressure, and QT intervals in normotensive obese women. AB - OBJECTIVE: We describe associations among the heart-rate-corrected QT (QTc) interval, QTc dispersion (QTc-d), circadian BP variation, and autonomic function in obese normotensive women and the effect of sustained weight loss. RESEARCH METHODS AND PROCEDURES: In 71 obese (BMI = 37.14 +/- 2.6 kg/m(2)) women, 25 to 44 years of age, circadian BP variations (24-hour ambulatory BP monitoring), autonomic function (power spectral analysis of RR interval oscillations), and cardiac repolarization times (QTc-d and QTc interval) were recorded at baseline and after 1 year of a multidisciplinary program of weight reduction. RESULTS: Compared with nonobese age-matched women (n = 28, BMI = 23 +/- 2.0 kg/m(2)), obese women had higher values of QTc-d (p < 0.05) and QTc (p < 0.05), an altered sympathovagal balance (ratio of low-frequency/high-frequency power, p < 0.01), and a blunted nocturnal drop in BP (p < 0.01). In obese women, QTc-d and the QTc interval correlated with diastolic nighttime BP (p < 0.01) and sympathovagal balance (p < 0.01). Waist-to-hip ratio, free fatty acids, and plasma insulin levels correlated with QT intervals and reduced nocturnal drops in both systolic and diastolic BP and sympathovagal balance (p < 0.01). After 1 year, obese women lost at least 10% of their original weight, which was associated with decrements of QTc-d (p < 0.02), the QTc interval (p < 0.05), nighttime BP (p < 0.01), and sympathovagal balance (p < 0.02). DISCUSSION: Sustained weight loss is a safe method to ameliorate diastolic nighttime BP drop and sympathetic overactivity, which may reduce the cardiovascular risk in obese women. PMID- 12740456 TI - Assessment of global coronary heart disease risk in overweight and obese African American women. AB - OBJECTIVE: To examine, with the use of national guidelines, coronary heart disease (CHD) risk with increasing BMI for primary prevention in urban African American women. RESEARCH METHODS AND PROCEDURES: Participants were recruited for CHD risk factor screening from 20 churches as part of a larger study of nutrition and fitness (Project Joy). All participants had a demographic, smoking and medical history assessment, and the following measurements were taken: weight, height, waist circumference, blood pressure, lipid levels, and glucose. Three methods of defining risk, the Framingham Point Scoring System, a count of risk factors, and the presence of the multiple metabolic syndrome, based on the National Cholesterol Education Program Adult Treatment Panel III Report and BMI classes established by the Clinical Guidelines, were used. RESULTS: A total of 396 women were eligible. Participants were 40 to 80 years of age and had marked excess prevalence of overweight and obesity (84%); 55% were obese. There was a linear increase in risk factors as BMI increased. Lipids did not differ significantly among BMI classifications. Seventeen percent of women had multiple metabolic syndrome. Eight percent and 16% of women in the normal and overweight BMI classes, respectively, had two or more modifiable risk factors. There was no difference in number of modifiable risk factors among the obese classes. The Framingham Point Scoring System assigned a <10% risk of a hard CHD event in 10 years to 97% of the women. DISCUSSION: National risk assessment guidelines for primary prevention of CHD may not be adequate for overweight and obese urban African-American women and require further study. PMID- 12740457 TI - Geographic and demographic variation in the prevalence of overweight Canadian children. AB - OBJECTIVE: To examine the geographic and demographic variation in the prevalence of overweight Canadian children. RESEARCH METHODS AND PROCEDURES: Using BMI data from the 1981 Canada Fitness Survey and the 1996 National Longitudinal Survey of Children and Youth, this study assessed: 1). the prevalence of overweight and obesity among Canadian boys and girls ages 7 to 13 years; 2). secular trends in the prevalence of overweight from 1981 to 1996, by province and adjusted for age and sex; and 3). provincial variation in the prevalence of overweight, before and after adjusting for socioeconomic and demographic characteristics. RESULTS: The prevalence of boys and girls classified as overweight in 1996 was 33% and 26%, respectively. The corresponding figures for obesity were 10% for boys and 9% for girls. Provincial variation was observed with a trend of increasing risk of being overweight from west to east. Socioeconomic status was inversely related to the prevalence of overweight regardless of geographic region. The risk of being overweight was more related to geography (province) than demographic variables (income and family background); however, the effect of secular trends (1981 to 1996) exceeded the effect of geographic or demographic variables. DISCUSSION: The prevalence of childhood overweight and obesity is increasing in all areas of Canada and can be explained only partially by geographic or demographic characteristics. PMID- 12740458 TI - Regional fat pad growth and cellularity in obese zucker rats: modulation by caloric restriction. AB - OBJECTIVE: To investigate, in young obese male Zucker rats, the effects of chronic food restriction and subsequent refeeding on: 1). parameters of nonadipose and adipose growth, 2). regional adipose depot cellularity [fat cell volume (FCV) and number], and 3). circulating leptin levels. RESEARCH METHODS AND PROCEDURES: Obese (fa/fa) and lean (Fa/?) male Zucker rats were studied from age 5 to 19 weeks. After baseline food intake monitoring, 10 obese rats were subjected to 58 days of marked caloric restriction from ad libitum levels [obese restricted (OR)], followed by a return to ad libitum feeding for 22 days. Ten lean control rats and 10 obese control rats were fed ad libitum for the entire experiment. All rats were fed using a computer-driven automated feeding system designed to mimic natural eating patterns. RESULTS: After food restriction, OR rats weighed significantly less than did lean and obese rats and showed a significant diminution in body and adipose growth as compared with obese rats. Relative adiposity was not different between obese and OR rats and was significantly higher than that of lean rats. The limitation in growth of the adipose tissue mass in OR rats was due mostly to suppression of fat cell proliferation because the mean FCV in each of the four depots was not affected. Serum leptin levels of OR and obese rats were not different from each other but were significantly higher than those of lean rats. DISCUSSION: Marked caloric restriction affects obese male Zucker rats in a manner different from that of nongenetic rodent models (i.e., Wistar rats). In comparison with the response to caloric deprivation of Wistar rats, these calorically restricted obese male Zucker rats appeared to defend their relative adiposity and mean FCV at the expense of fat cell number. These findings indicate that genetic and/or tissue specific controls override the general consequences of food restriction in this genetic model of obesity. PMID- 12740459 TI - Chitosan supplementation and fecal fat excretion in men. AB - OBJECTIVE: Few weight loss supplements are clinically tested for efficacy, yet their proliferation continues. Chitosan-based supplements are sold as fat trappers and fat magnets. They purportedly block fat absorption and cause weight loss without food restriction. We quantified the in vivo effect of a chitosan product on fat absorption. RESEARCH METHODS AND PROCEDURES: Participants (n = 15) consumed five meals per day for 12 days. Energy intake was not restricted. Participants consumed no supplements during a 4-day control period and two capsules five times per day (4.5 g chitosan/d), 30 minutes before each meal, during a 4-day supplement period. All feces were collected from days 2 to 12. Oral charcoal markers permitted division of the feces into two periods. The two fecal pools were analyzed for fat content. RESULTS: Participants were male, 26.3 +/- 5.9 years old, BMI of 25.6 +/- 2.3 kg/m(2). Subjects consumed 133 +/- 23 g of fat/d and 12.91 +/- 1.79 MJ/d (3084 +/- 427 kcal/d). Individual meals averaged 26.3 +/- 9.3 g of fat. With chitosan supplementation at 10 capsules/day, fecal fat excretion increased by 1.1 +/- 1.8 g/d (p = 0.02), from 6.1 +/- 1.2 to 7.2 +/ 1.8 g/d. DISCUSSION: The effect of chitosan on fat absorption is clinically negligible. Far from being a fat trapper, at 0.11 +/- 0.18 g of fat trapped per 0.45-g capsule or 1.1 g (9.9 kcal) fat trapped per day, this product would have no significant effect on energy balance. The fat trapping claims associated with chitosan are unsubstantiated. PMID- 12740460 TI - Pediatric cervical spine: normal anatomy, variants, and trauma. AB - Emergency radiologic evaluation of the pediatric cervical spine can be challenging because of the confusing appearance of synchondroses, normal anatomic variants, and injuries that are unique to children. Cervical spine injuries in children are usually seen in the upper cervical region owing to the unique biomechanics and anatomy of the pediatric cervical spine. Knowledge of the normal embryologic development and anatomy of the cervical spine is important to avoid mistaking synchondroses for fractures in the setting of trauma. Familiarity with anatomic variants is also important for correct image interpretation. These variants include pseudosubluxation, absence of cervical lordosis, wedging of the C3 vertebra, widening of the predental space, prevertebral soft-tissue widening, intervertebral widening, and "pseudo-Jefferson fracture." In addition, familiarity with mechanisms of injury and appropriate imaging modalities will aid in the correct interpretation of radiologic images of the pediatric cervical spine. PMID- 12740462 TI - Comprehensive imaging of ischemic stroke with multisection CT. AB - Computed tomography (CT) is an established tool for the diagnosis of ischemic or hemorrhagic stroke. Nonenhanced CT can help exclude hemorrhage and detect "early signs" of infarction but cannot reliably demonstrate irreversibly damaged brain tissue in the hyperacute stage of ischemic stroke. Further evaluation of patients with ischemic stroke should include differentiation between reversible and irreversible brain damage, which is essential for choosing an appropriate therapy. Perfusion CT provides information about brain perfusion, which permits differentiation of irreversibly damaged brain tissue from reversibly impaired "tissue at risk." CT angiography can help detect stenosis or occlusion of extra- and intracranial arteries. Multisection CT allows the combined use of all three imaging modalities-nonenhanced CT, perfusion CT, and CT angiography-to rapidly obtain comprehensive information regarding the extent of ischemic damage in acute stroke patients. Specific patterns of findings are typically seen in ischemic stroke and can be analyzed more accurately with the combined use of multisection CT and MR imaging. Nevertheless, prospective studies involving a large number of patients will be needed to ascertain the treatment of choice for patients with each of these patterns of findings. PMID- 12740463 TI - Extensor mechanism of the fingers: MR imaging-anatomic correlation. AB - Extension of the fingers is a complex function carried out by simultaneous action of extrinsic and intrinsic muscles, as well as retinacular structures in the dorsum of the wrist, hand, and fingers that support and coordinate the action of the muscles. The extensor mechanism of the fingers is divided into topographic zones, which extend from the forearm to the distal phalanx. Magnetic resonance (MR) imaging shows in detail the musculotendinous and retinacular structures of the extensor apparatus. In the different extensor zones, MR imaging findings are similar to those seen macroscopically in anatomic sections. Understanding of and familiarity with the extensor anatomy of the hand and fingers by the radiologist is crucial for better assessment of pathologic conditions with MR imaging and optimization of this modality as a diagnostic tool. Extensor tendon injuries and tenosynovitis represent clinical situations in which knowledge of this anatomy is useful for the clinical radiologist. PMID- 12740464 TI - Imaging of foot and ankle nerve entrapment syndromes: from well-demonstrated to unfamiliar sites. AB - Nerve entrapment at the foot and ankle involves thin and complex anatomic structures and is underdiagnosed because clinical symptoms and electrophysiologic findings may not contribute to the diagnosis. Nerve entrapment can be secondary to acute trauma or repetitive microtrauma. The latter often results from intensive sports-related activity, inappropriate footwear, or internal foot derangement. Various lesions that occur in fibro-osseous tunnels can cause nerve compression (eg, ganglion cysts, varicosities, bone and joint abnormalities, tumors, tenosynovitis, supernumerary or hypertrophic muscles). Accurate nerve examination must be performed, particularly in patients with atypical ankle pain, to detect focal tenderness or paresthesia. Ultrasonography is useful in this setting because it yields both clinical and morphologic findings. High-resolution magnetic resonance imaging provides accurate delineation of the nervous system anatomy. Furthermore, technologic developments in the field of radiology are making it possible to obtain clearer, more accurate images. Radiologists must be aware of the main nerve entrapment syndromes at the foot and ankle and be able to perform accurate nerve examinations with different imaging modalities in patients with foot and ankle pain. PMID- 12740465 TI - Dedicated multidetector CT of the stomach: spectrum of diseases. AB - Multidetector computed tomography (CT) offers new opportunities in imaging of the gastrointestinal tract. When thin collimation is used, near-isotropic imaging of the stomach is possible, allowing high-quality multiplanar reformation and three dimensional reconstruction of gastric images. Proper distention of the stomach and optimally timed administration of intravenous contrast material are required to detect and characterize disease. In contrast to gastroscopy and double contrast studies of the stomach, CT provides information about both the gastric wall and the extragastric extent of disease. Preoperative staging of gastric carcinoma appears to be the main clinical indication for multidetector CT. In addition, multidetector CT allows detection of other gastric malignancies (lymphoma, carcinoid tumors, metastases, gastrointestinal stromal tumors) and benign gastric tumors (neural tumors, polyps). Gastric inflammation (gastritis, ulcers, Menetrier disease) and miscellaneous gastric conditions (emphysema, gastric outlet obstruction, varices) can also be visualized with multidetector CT. Multidetector CT is a valuable tool for the evaluation of gastric wall disease and serves as an adjunct to endoscopy. PMID- 12740466 TI - Primary neoplasms of the appendix: radiologic spectrum of disease with pathologic correlation. AB - Although uncommon, primary appendiceal neoplasms often result in clinical symptoms that may lead to abdominal imaging. Acute appendicitis from luminal obstruction is the most common manifestation for most tumor types. Other manifestations include intussusception, a palpable mass, gastrointestinal bleeding, increasing abdominal girth (from pseudomyxoma peritonei), and secondary genitourinary complications. Asymptomatic appendiceal neoplasms may be discovered incidentally. Mucoceles from either benign or malignant mucinous neoplasms represent the majority of appendiceal tumors detected at imaging but are the least likely to manifest as appendicitis. Pseudomyxoma peritonei is a common manifestation of mucinous adenocarcinoma. Colonic-type (nonmucinous) adenocarcinoma of the appendix is much less common than mucinous tumors and typically manifests as a focal mass without mucocele formation. Carcinoid tumor is the most common appendiceal neoplasm but is less often detected radiologically because it is typically small and relatively asymptomatic. Goblet cell carcinoid tumor and non-Hodgkin lymphoma of the appendix are rare and usually infiltrate the entire appendix. Cross-sectional imaging, particularly computed tomography (CT), is effective in the evaluation of these neoplasms. CT appears to be the modality of choice whenever an appendiceal mass is suspected. CT will help rule out or confirm an appendiceal tumor and may suggest a more specific diagnosis. PMID- 12740467 TI - US of the peritoneum. AB - Familiarity with the pathophysiology of peritoneal disease is the basis of successful ultrasound (US) study of the peritoneum. The pouch of Douglas, diaphragmatic surfaces, the paracolic gutters, and the regions of the mesentery and omentum should receive careful scrutiny in the patient at risk for a peritoneal disease process. An optimal US technique requires assessment of the entire peritoneum with a transducer selected to reflect the depth of the region of interest. US may demonstrate minute quantities of free intraperitoneal fluid and is therefore capable of providing sensitive quantitative information about ascites. Qualitative information may also be inferred, as blood, pus, and neoplastic cells demonstrate correlation with particulate ascites on gray-scale US scans. Peritoneal nodules, plaques, and thickening may be detected on the visceral or parietal peritoneal surfaces, especially when high-frequency probes are used. Transvaginal study in women increases the sensitivity of US for detection of peritoneal disease. In women who have unexplained sepsis or are at risk for carcinomatosis, transvaginal scanning should routinely be added to the regular abdominal and pelvic studies regardless of the findings of those studies. Peritoneal carcinomatosis, primary peritoneal neoplasms, pseudomyxoma peritonei, and peritonitis have characteristic appearances at US. PMID- 12740469 TI - "Milk of calcium" and food signs in radiology. PMID- 12740470 TI - Multidetector CT of peritoneal carcinomatosis from ovarian cancer. AB - Ovarian cancer is usually in an advanced stage at diagnosis due to the presence of peritoneal carcinomatosis, which develops as a result of peritoneal fluid circulation. Tumor implants of varying size can occur anywhere from the diaphragm through the pelvis. Computed tomography (CT) can be used to detect these metastatic lesions, which can be miliary or large and appear as soft-tissue or low-attenuation masses. Recent advances in CT technology have increased the flexibility of image acquisition, thereby allowing the use of thin sections and multiplanar reformatting. With multidetector CT, thin-section images of the abdomen and pelvis can be obtained to assess for subcentimeter implants and to create three-dimensional images with reduced artifact. Multiplanar reformatting can be used to confirm the presence of implants. Structures such as the diaphragm, paracolic gutters, bowel, and cul-de-sac can be evaluated in multiple planes for surface nodularity and small implants. Interactive multiplanar review of the abdomen and pelvis has the potential to improve detection of peritoneal metastases at CT. PMID- 12740471 TI - US of abnormal uterine bleeding. AB - Any significant deviation from a woman's established menstrual pattern may be considered abnormal uterine bleeding, and several factors direct evaluation of a patient with such bleeding. Premenopausal disorders that are well evaluated with ultrasound (US) include endometriosis, adenomyosis, and leiomyomas. A positive pregnancy test in a woman of childbearing age prompts a search for an intrauterine pregnancy. Possible complications that may contribute to bleeding include ectopic pregnancy; placental factors including position, trauma, and infection; gestational trophoblastic disease; preterm labor; and postpartum complications. Atrophic changes, hormonal status, and carcinoma are considerations in the postmenopausal patient with abnormal uterine bleeding. Foreign bodies, trauma, infection, polyps, and iatrogenic causes can be observed in all groups. Differential diagnoses for abnormal uterine bleeding in premenopausal, pregnant, and postmenopausal patients are well evaluated with US, and US techniques have greatly facilitated evaluation of pelvic disease. Urgent and emergent conditions such as ectopic pregnancy, placenta previa, and preterm labor are readily identifiable. PMID- 12740472 TI - Inflammatory pseudotumor. AB - Inflammatory pseudotumor is a quasineoplastic lesion that most commonly involves the lung and the orbit, but it has been reported to occur in nearly every site in the body. The pathogenesis, natural history, clinical presentation, imaging findings, and treatment options for inflammatory pseudotumor in the lung, heart, gastrointestinal tract, adrenal gland, iliopsoas muscle, orbit, and central nervous system are discussed. Because inflammatory pseudotumors mimic malignant tumors both clinically and radiologically, the radiologist should be familiar with this entity and help avoid unnecessary radical surgery when possible. PMID- 12740473 TI - Foreign bodies. AB - Foreign bodies are uncommon, but they are important and interesting. Foreign bodies may be ingested, inserted into a body cavity, or deposited into the body by a traumatic or iatrogenic injury. Most ingested foreign bodies pass through the gastrointestinal tract without a problem. Most foreign bodies inserted into a body cavity cause only minor mucosal injury. However, ingested or inserted foreign bodies may cause bowel obstruction or perforation; lead to severe hemorrhage, abscess formation, or septicemia; or undergo distant embolization. Motor vehicle accidents and bullet wounds are common causes of traumatic foreign bodies. Metallic objects, except aluminum, are opaque, and most animal bones and all glass foreign bodies are opaque on radiographs. Most plastic and wooden foreign bodies (cactus thorns, splinters) and most fish bones are not opaque on radiographs. All patients should be thoroughly screened for foreign bodies before undergoing a magnetic resonance imaging study. PMID- 12740474 TI - From the archives of the AFIP: Localized fibrous tumor of the pleura. AB - Eighty-two localized fibrous tumors of the pleura (LFTP) were reviewed retrospectively for the clinical, pathologic, and radiologic findings. Forty-four women and 38 men ranged in age from 17 to 78 years (mean, 54.7 years). Sixty-four benign LFTP ranged in size from 2 to 30 cm (mean, 13.2 cm), and 18 malignant tumors ranged from 3 to 23 cm (mean, 14.4 cm). Forty-eight patients (60%) presented with symptoms. Radiographs of 76 patients demonstrated solitary masses occupying or extending into the inferior hemithorax (79%). Computed tomography (CT) of 78 lesions demonstrated lobular masses (83%) that formed at least one acute angle (96%) or only acute angles (65%) with the adjacent pleura. Heterogeneous lesion attenuation was documented in 88% of enhanced and in 68% of unenhanced CT scans. Contrast enhancement was common (62% of cases). Magnetic resonance (MR) imaging of 18 lesions demonstrated heterogeneous signal intensity on both T1- and T2-weighted images (78% and 83%, respectively). Multiplanar MR imaging allowed visualization of the diaphragm and documentation of an intrathoracic mass in all cases. LFTP are solitary lobular heterogeneous masses that occur in symptomatic adults and often affect the inferior hemithorax. Malignant lesions are radiologically indistinguishable from those with benign histologic characteristics. Radiographic and CT features characteristic of pleural location are typically absent. PMID- 12740476 TI - Best cases from the AFIP: Involuted sclerotic meningioma. PMID- 12740477 TI - Malignant peripheral nerve sheath tumor. PMID- 12740478 TI - PACSPulse: a web-based DICOM network traffic monitor and analysis tool. AB - PACSPulse, an open-source tool, was developed to identify and analyze the performance bottlenecks of picture archiving and communication systems (PACS). PACSPulse provides a graphical Web interface for straightforward analysis of PACS performance on the basis of data acquired by tracking usage by network, server, workstation, type of traffic, and time of day. The PACS archive logs performance and usage data on image traffic being sent to it from the imaging units and study data requested by users. The performance log is sent via file transfer protocol (FTP) to a separate server for analysis. The data are parsed and sent to a database server connected to a Web server. The Web site is used to depict trends in the performance of the entire system to detect signs of degradation. The system was built entirely of open-source components for the operating system, database, charting tool, and Web server. Performance monitoring is an essential tool for analyzing, understanding, and predicting the performance characteristics of a PACS. PMID- 12740479 TI - Inflammatory reactions in the pathogenesis of atherosclerosis. AB - Atherosclerosis and its complications constitute the most common causes of death in Western societies and Japan. Although several theories or hypotheses about atherogenesis have been proposed during the past decades, none can completely explain the whole process of the pathogenesis of atherosclerosis because this disease is associated with multiple risk factors. In spite of this, the concept that atherosclerosis is a specific form of chronic inflammatory process resulting from interactions between plasma lipoproteins, cellular components ( monocyte/macrophages, T lymphocytes, endothelial cells and smooth muscle cells ) and the extracellular matrix of the arterial wall, is now well accepted. Histologically, atherosclerotic lesions from the early-stage ( fatty streak ) to more complicated lesions possess all the features of chronic inflammation. It has been demonstrated that atherogenic lipoproteins such as oxidized low density lipoprotein ( LDL ), remnant lipoprotein (beta-VLDL) and lipoprotein [ Lp ] ( a ) play a critical role in the pro-inflammatory reaction, whereas high density lipoprotein ( HDL ), anti-atherogenic lipoproteins, exert anti-inflammatory functions. In cholesterol-fed animals, the earliest events in the arterial wall during atherogenesis are the adhesion of monocytes and lymphocytes to endothelial cells followed by the migration of these cells into the intima. It has been shown that these early events in atherosclerosis are triggered by the presence of high levels of atherogenic lipoproteins in the plasma and are mediated by inflammatory factors such as adhesion molecules and cytokines in the arterial wall. The development of genetically modified laboratory animals ( transgenic and knock-out mice and transgenic rabbits ) has provided a powerful approach for dissecting individual candidate genes and studying their cause-and-effect relationships in lesion formation and progression. The purpose of this article is to review the recent progress regarding the inflammatory processes during the development of atherosclerosis based on both human and experimental studies. In particular, we will address the mechanisms of atherogenic lipoproteins in terms of inflammatory reactions associated with hypercholesterolemia. Understanding the molecular mechanisms responsible for inflammatory reactions during atherogenesis may help us to develop novel therapeutic strategies to control, treat and prevent atherosclerosis in the future. PMID- 12740480 TI - Relationship between high-density lipoprotein-cholesterol and malondialdehyde modified low-density lipoprotein concentrations. AB - Several reports have suggested that HDL has anti-oxidative actions. We investigated the relationship between HDL-cholesterol (HDL-C) and malondialdehyde modified LDL (MDA-LDL) concentrations using enzyme linked immunosolvent assay. We divided our study subjects into four groups on the basis of concentrations of triglyceride (TG) and HDL-C by the following lipid profiles: serum TG < or = 1.69 mmol/L and HDL-C > or = 1.16 mmol/L (control group, n = 26); TG >1.69 and HDL-C < or = 1.16 (high TG group, n = 22); TG >1.69 and HDL-C < or = 0.91 (high TG & low HDL group, n = 67); TG < or = 1.69 and HDL-C < or = 0.91 (low HDL group, n = 21). MDA-LDL concentrations, MDA-LDL/apolipoprotein B (apo B) ratio, and LDL size were different between subjects in high TG & low HDL and control groups. MDA-LDL concentrations in both high TG and low HDL groups did not differ significantly from those in the control. However, MDA-LDL/apo B ratio in low HDL group was significantly higher than that in the control (P < 0.05). The MDA-LDL/apo B ratio reflects the extent of MDA modification of apo B in LDL. Therefore, our data suggest that as HDL-C concentrations fall, the extent of MDA modification per one LDL particle increases. Moreover, accompanied by high TG concentration, LDL size in subjects with lower HDL-C concentrations became smaller. PMID- 12740481 TI - Apolipoprotein H variant modifies plasma triglyceride phenotype in familial hypercholesterolemia: a molecular study in an eight-generation hyperlipidemic family. AB - In the course of investigating familial coronary artery disease in Utah, we studied 196 members of an eight-generation extended family of familial hypercholesterolemia (FH), in which 73 members were affected with type IIa hyperlipoproteinemia (HLPIIa; high plasma cholesterol) and 11 members with type IIb hyperlipoproteinemia (HLPIIb; high plasma cholesterol as well as plasma triglyceride). A splice-site mutation of the LDL receptor (LDLR) gene (IVS14 + G > A) co-segregated with elevated plasma cholesterol among all the members, but not with the elevated plasma triglyceride and VLDL cholesterol levels seen in HLPIIb patients. The apolipoprotein H (apoH) gene plays a role in plasma triglyceride removal and lipoprotein lipase enhancement. Intra-familial correlation analysis of the modifier effect of Val247Leu substitution in the apoH gene was carried out among 84 LDLR-mutation carriers and 112 non-carriers. When plasma triglyceride levels in the LDLR-mutation carriers were compared, the values were lowest among V/V homozygotes (mean +/- SD = 145 +/- 53 mg/dl), highest in L/L homozygotes (277 +/- 177 mg/dl), and intermediate among V/L heterozygotes (191 +/- 102 mg/dl) (p = 0.0015). All eleven patients who presented with HLPIIb had inherited both the defective LDLR allele and an apoH 247Leu allele, whereas all 45 carriers of the defective LDLR allele not carrying the apoH Leu allele presented with HLPIIa but not HLPIIb (p = 0.0001). These results indicate a significant modification of the phenotype of FH with a defective LDLR allele, by apoH Leu variation in our studied family. PMID- 12740482 TI - R/R genotype of human paraoxonase (PON1) is more protective against lipoprotein oxidation and coronary artery disease in Japanese subjects. AB - Human paraoxonase (PON1) is an high-density lipoprotein (HDL) -associated enzyme that is proposed to protect against the oxidation of lipoproteins. Recently, the association of coronary artery disease (CAD) and PON1 activity was reported. Furthermore, the R/R genotype of PON1 has been related to the risk for CAD. In this study we investigated the PON1 genotype and susceptibility to lipoprotein oxidation to elucidate the contribution of PON1 to atherosclerosis in Japanese subjects. We studied 179 patients who underwent coronary angiography and their PON1 genotypes were determined. Lipoproteins were obtained from a patient's blood after at least 12 hours fasting and were separated with sequential ultracentrifugation. We analyzed the thiobarbituric acid reactive substances (TBARS) and continuously monitored the copper-induced oxidation three genotype groups. Genotype frequencies of Q/Q, Q/R, and R/R were 21.2%, 36.9%, and 41.9%, respectively. PON1 polymorphism clearly determined the lipid oxidation. The R/R genotype of PON1 had significantly lower levels of plasma and HDL TBARS and significantly retarded the initiation of oxidation in HDL and low-density lipoprotein (LDL). The R/R genotype was related to the lower prevalence of CAD. The PON1 genotype clearly determined the oxidative modification of lipoproteins and may play a role in the pathogenesis of atherosclerosis via its protective effect against lipoprotein oxidation in Japanese subjects. PMID- 12740483 TI - Effect of cilostazol on impaired vasodilatory response of the brachial artery to ischemia in smokers. AB - The vascular endothelial function of smokers is known to be impaired. This study investigated whether cilostazol could improve the vasodilatory response of the brachial artery to ischemia, an indicator of endothelial function, in ten male smokers. Endothelium-dependent vasodilatation and endothelium-independent vasodilatation of the brachial artery were measured in 11 male non-smokers and 20 male smokers with matching age and weight. The results showed that the vasodilatory response to reactive hyperemia was significantly smaller in the smokers (4.8 +/- 1.6%) when compared to that in the non-smokers (7.6 +/- 2.5%) (p = 0.0013). However, no significant difference in the vasodilatory response to isosorbide dinitrate was observed between the two groups. In addition, there were no significant differences in serum lipid, Lp (a), or blood homocysteine between the smokers and non-smokers. When 150 mg/day of cilostazol was administered for two weeks, the vasodilatory response to reactive hyperemia significantly improved (4.2 +/- 1.2% to 7.8 +/- 3.5%, p = 0.0032). The increased vasodilatory response to reactive hyperemia by cilostazol was reduced after cessation of the drug (4.5 +/- 1.5%). These findings suggest that cilostazol improves vascular endothelial dysfunction in smokers. PMID- 12740484 TI - Apoptosis of endothelial cells may be mediated by genes of peroxisome proliferator-activated receptor gamma 1 (PPARgamma 1) and PPARalpha genes. AB - Apoptosis in human umbilical vein endothelial cells (HUVECs) was prevented by transfection with the gene for the human full-length peroxisome proliferator activated receptor alpha (PPARalpha), or acyl-coenzyme A synthetase (AcylCS) into HUVECs. In contrast, ligands/activators of PPARgamma 1 induced apoptosis by a cytochrome c-dependent mechanism in HUVECs transfected with human full-length PPARgamma 1, but not in hepatocytes. Co-transfection of PPARgamma 1 and PPARalpha protected the HUVEC apoptosis. The results suggest that the apoptosis of endothelial cells may be mediated by genes of PPARgamma1 and PPARalpha. PMID- 12740485 TI - Plaque-stabilizing effect of pitavastatin in Watanabe heritable hyperlipidemic (WHHL) rabbits. AB - This study investigated the effect of pitavastatin, a 3-hydroxy-3-methylglutaryl coenzyme A ( HMG-CoA ) reductase inhibitor with strong cholesterol-lowering activity, on the composition of atherosclerotic plaque. Pitavastatin ( 0.5mg/kg ) was administered to Watanabe heritable hyperlipidemic ( WHHL ) rabbits for 16 weeks, with the result that plasma total cholesterol ( TC ), very low density lipoprotein ( VLDL )-C, intermediate density lipoprotein ( IDL )-C and low density lipoprotein ( LDL )-C decreased by 28.6, 60.0, 42.3 and 21.7%, respectively. In the aorta, pitavastatin reduced the area of the lesion by 38.6%. In the pitavastatin group, the macrophage-positive area in the aortic plaque was reduced by 39.4%, and the areas occupied by collagen and a-smooth muscle actin ( alpha-SMA )-positive area increased by 66.4 and 91.7%, respectively. In the aortic arch, pitavastatin increased the average thickness of alpha-SMA in the plaque by 96.7% and reduced the vulnerability index by 76.0%. Furthermore, pitavastatin reduced the positive areas of monocyte chemoattractant protein ( MCP )-1, matrix metalloproteinase ( MMP )-3 and MMP-9 by 39.1, 40.6 and 52.3%, respectively. These results indicated that pitavastatin had an excellent lipid lowering effect in WHHL rabbits, suppressing the progression of atherosclerosis and stabilizing atherosclerotic plaque. PMID- 12740486 TI - Contribution of Rho A and Rho kinase to platelet-derived growth factor-BB-induced proliferation of vascular smooth muscle cells. AB - In order to identify small G protein (s) which contributes to the proliferation of vascular smooth muscle cells (VSMCs), we examined the effect of an HMG-CoA reductase inhibitor (cerivastatin), a farnesyltransferase inhibitor (FTI-277), a geranyl geranyl transferase inhibitor (GGTI-286) and a Rho kinase inhibitor (Y 27632) on the proliferation of cultured rat VSMCs stimulated with 20ng/ml platelet-derived growth factor (PDGF)-BB. Cerivastatin and GGTI-286, but not FTI 277, suppressed the PDGF-BB-induced activation of extracellular signal related kinase (ERK1/2). The inhibitory effect of cerivastatin on the PDGF-BB-induced activation of ERK1/2 was fully recovered by the addition of geranylgeranyl pyrophosphate (GGPP), but not farnesyl pyrophosphate (FPP). Cerivastatin and GGTI 286, but not FTI-277, suppressed the PDGF-BB-induced [3H] thymidine incorporation and activation of ornitine decarboxylase (ODC), both of which were fully recovered by the addition of GGPP, but not FPP. These data indicate that the PDGF BB-induced activation of ERK1/2 and proliferation of VSMCs depend upon geranylgeranylated small G protein. Immunoblotting analysis revealed the upregulation of Rho A protein in the membrane fractions of VSMCs stimulated by PDGF-BB. Furthermore, Y-27632 suppressed the PDGF-BB-induced activation of ERK1/2 and proliferation of VSMCs. On the basis of these data, we conclude that PDGF-BB stimulates the proliferation of VSMCs via the activation of Rho A. Rho kinase plays an important role in this process as an effector of Rho A. PMID- 12740487 TI - Expeditious method of urethrovesical junction determination in retropubic colposuspension with intraballoon illumination of Foley catheter. AB - OBJECTIVE: To investigate the efficiency and safety of Foley catheter intraballoon illumination in determining the urethrovesical junction (UVJ) in retropubic colposuspension. METHOD: Our accessory set for UVJ determination is composed of a 24-french Foley catheter and a 5-mm 0 degrees telescope. The telescope is inserted into the catheter from the urinary luminal side until the tip has reached the level of the catheter balloon. After dissection of the bladder, urethra, and paraurethral tissues to reveal the submucosal endopelvic fascia, the accessory set is inserted into the bladder and the balloon is inflated with 5 ml air and illuminated by a switch on the cold light source. To determine the right side of UVJ, the catheter is pulled slightly downward and to the right of the patient. The contralateral side of the UVJ is determined by pulling the catheter slightly downward and to the left of the patient. RESULT: Intraballoon illumination of the Foley catheter helps to ensure accurate placement of paraurethral sutures at UVJ. CONCLUSION: Intraballoon illumination for determination of UVJ is simple, safe, effective and precludes an invasive approach such as cystoscopy and cystotomy. PMID- 12740488 TI - Evaluation of fragmentation with single or multiple pulse setting of Lithoclast for renal calculi during percutaneous nephrolithotripsy and its impact on clearance. AB - OBJECTIVES: To evaluate the impact of single or multiple pulse settings of Lithoclast to fragment renal calculi, and its effect on fragment size and clearance. MATERIAL AND METHODS: In a prospective study 153 patients, who had been treated over a period of 35 months for renal stone disease by percutaneous nephrolithotomy, were evaluated. In 69 cases the single and in 84 cases the multiple pulse modes were used to fragment the stone intracorporeally using the pneumatic lithoclast. The 2 groups were similar with regard to patient characteristics, stone location and size. RESULTS: Most stones were staghorn. The lithoclast was effective with good fragmentation in all cases. The mean operating and fluoroscopy times in the single and multiple pulse mode were 124.1 and 5.8, and 141.2 and 7.4 min, respectively (both differences statistically significant, p < 0.001). Immediate postoperatively residual stone fragments were seen in 16 cases with the single and 35 cases with the multiple pulse mode setting (difference statistically significant, p < 0.05). Relook procedures were required in 14 cases with the single and 22 cases with the multiple pulse setting (difference not statistically significant). No specific complication was seen related to the use of either of the two modes. On follow-up 1 patient in the multiple pulse setting developed stone recurrence after 19 months. CONCLUSIONS: The single pulse mode was associated with controlled fragmentation of the stone, formation of larger fragments which were easier to pick up, less stone scatter, shorter operating time and less exposure to fluoroscopy. The single pulse mode was also associated with statistically less chances of postoperative residual fragments. PMID- 12740489 TI - Management and follow-up of impacted ureteral stones. AB - INTRODUCTION: Impacted stones are those that remain unchanged in the same location for at least 2 months. MATERIALS AND METHODS: We evaluated 42 patients with impacted ureteral stones, and followed them for two and a half years to check for long-term results. The calculi location included all three segments of the ureter (proximal, mid and distal). Patients' age ranged from 22 to 83 years (mean 52.5 years). Primarily, patients were manipulated with extracorporeal shock wave lithotripsy (ESWL) in situ, or following stenting. If the result was not satisfactory, then we proceeded to retrograde ureteroscopy and ureterolithotripsy. Open ureterolithotomy was our final choice. RESULTS: Thirty six of the 42 patients (85.7%) were stone-free without the need of an open procedure. Follow-up period ranged from 10 up to 40 months, with a median period of 30 months and was achieved in 30 patients (71.4%). Stone recurrence was noted in 4 cases, while hydronephrosis without evidence of stone presence in 2. CONCLUSIONS: The initial approach for the treatment of impacted lithiasis should be attempted by ESWL. If this fails, alternative therapeutic solutions such as endoscopy can result in removal of the stone. PMID- 12740490 TI - Metabolic risk factors in pediatric and adult calcium oxalate urinary stone formers: is there any difference? AB - OBJECTIVES: Urolithiasis in children is recognized with an increasing frequency, while exact etiological factors remain to be determined. The aim of this study is to compare the metabolic risk factors and saturation of urine in pediatric and adult calcium oxalate (Ca-Ox) stone formers. METHODS: A total of 33 pediatric (mean age: 6.8 +/- 3.1 years) and 120 adult patients (mean age: 39.7 +/- 5.7 years), with documented Ca-Ox urinary stone disease, underwent a comprehensive metabolic evaluation at our institution. Beside a broad serum analysis, concentrations of calcium, oxalate, magnesium, uric acid and citrate were measured in 24-hour collected urine. Saturation of urine was calculated by Marshall-Robertson's nomograms. RESULTS: Hypocitraturia, observed in 60.6%, and hypomagnesuria, detected in 39.4%, but not hypercalciuria, were the most common metabolic risk factors in the pediatric group. In adults, hypercalciuria still represented one of the major metabolic risk factors, detected in 44.1%, although hypocitraturia, observed in 45.8%, was the most prevalent metabolic risk factor, as it was in the pediatric group. Pediatric cases had significantly (p < 0.05) higher prevalence of hypocitraturia, hypomagnesuria and supersaturated urine when compared to adults. Metabolic abnormalities could be detected in a high percentage (82%) of primary and recurrent pediatric Ca-Ox stone formers, but not in primary adult stone formers. CONCLUSIONS: Metabolic risk factors significantly differ in pediatric and adult Ca-Ox stone formers. Hypocitraturia and hypomagnesuria seem to play a major role in stone formation, and metabolic abnormalities can be detected in a significant percentage of both primary and recurrent pediatric stone formers. Thus, a comprehensive metabolic evaluation is of utmost importance for all children with Ca-Ox stones. PMID- 12740491 TI - Association of E-cadherin gene 3'-UTR C/T polymorphism with calcium oxalate stone disease. AB - INTRODUCTION: Urinary stone disease is one of the most commonly seen urological diseases in Taiwan. Single nucleotide polymorphisms (SNPs) are commonly used for the investigation of genetic markers for stone disease. E-cadherin (CDH-1) is one of the cellular junction proteins related to the integrity of epithelial cells. Our aim was to investigate a polymorphism of the CDH-1 gene 3'-UTR as a possible genetic marker in the search for the genetic causes. MATERIALS AND METHODS: 148 patients with calcium oxalate stone were compared with 103 healthy controls for the frequency of CDH-1 3'-UTR polymorphisms. The polymorphism was detected by polymerase chain reaction-based restriction analysis (PML I endonuclease). RESULTS AND CONCLUSIONS: The results revealed significant differences between normal individuals and calcium stone disease patients (p = 0.0013). The distribution of genotype TT homozygote was higher in stone patients (51.5%) than in the control group (43.4%). The odds ratio for T allele compared to C allele was 2.0. We have concluded that polymorphisms of CDH-1 3'-UTR is a valid genetic marker for calcium stone disease. PMID- 12740492 TI - Results of cadaver kidney transplantation with right renal vein extension. AB - INTRODUCTION: Several methods to lengthen the renal vein of the allografts have been proposed in order to secure the surgeon's field of vision and to allow appropriate positioning of the graft. However, its safety and complications have not been clarified. Furthermore, its long- and short-term results have not been investigated. Therefore, we studied the results of this operative procedure to confirm the safety of the technique. METHODS: The right renal vein extension was utilized when the donated right kidney from a cadaver donor had sufficient length of the inferior vena cava wall. The results of 10 patients who received kidney transplantation with renal vein extension were examined. RESULTS: None of the patients had vascular complications. The transplanted kidney functioned in all patients; 3- and 5-year graft survival was 100 and 75.0%, respectively. CONCLUSIONS: The results confirmed that this technique was safe and without complications and did not affect the long- and short- term results of patients who received cadaver kidneys. Therefore, we conclude that it is appropriate to adapt this technique when the physician judges the long renal vein suitable for the procedure. PMID- 12740493 TI - Predisposing factors and treatment outcome in Fournier's gangrene. Analysis of 28 cases. AB - OBJECTIVES: To evaluate the etiologic factors and the effects of surgical debridement and adjunctive therapies on morbidity and mortality of Fournier's gangrene. METHODS: 27 males, 1 female, a total of 28 patients with a mean age of 58 years treated for Fournier's gangrene were evaluated retrospectively. RESULTS: Predisposing factors including diabetes, alcohol abuse, paraplegia and renal insufficiency were identifiable in 54% of the patients. Etiologic origin of the gangrene was urogenital, cutaneous and anorectal in 43, 25 and 11% of the patients, respectively. The pathology was limited to genitalia in 10, extending to perineum in 8, the umbilicus in 7 and even up to the axilla in 3 patients. Suprapubic cystostomy and colostomy were necessary in 18 and 2 cases, respectively. We used hyperbaric oxygen therapy in 2 and honey in 6 patients to accelerate wound healing. A repeat debridement was necessary in 39% of the cases. Plastic surgery and grafting were done in 14 patients. Our mortality rate was 7%. CONCLUSION: Early recognition of the pathology and aggressive surgical debridement are the mainstay of the management of Fournier's gangrene. Additional strategies to improve wound healing and increase patient survival are also needed. PMID- 12740494 TI - Prognostic consequences of the location of positive surgical margins in organ confined prostate cancer. AB - INTRODUCTION: The purpose of the present study was to evaluate and compare the risk of progression in organ-confined prostate cancers (stage pT2), according to the location of positive surgical margins. MATERIALS AND METHODS: Between 1988 and 2001, 538 consecutive men underwent radical prostatectomy for localized prostate cancer. All patients had preoperative physical examinations, serum PSA assays (Hybritech assay, N.l. <4 ng/ml) and ultrasound-guided sextant biopsies to confirm diagnosis. Radical prostatectomy specimens were analyzed according to the Stanford protocol. Positive margins were classified as single or multiple and main locations (apex, bladder neck and posterolateral) were noted. Postoperative follow-up data were obtained through routine serum PSA assays. Biochemical recurrence was defined as a single postoperative PSA level >0.2 ng/ml. Biochemical progression was studied in patients with organ-confined tumors (stage pT2) according to the location of the single positive margin. Kaplan-Meier analysis was performed to determine the actuarial biochemical recurrence-free likelihood and the log-rank test was used for statistical analysis. Differences were considered significant when the p value was <0.05. RESULTS: 371 patients had organ-confined tumors, and 60 patients (16.1%) had solitary positive margins (apex 26, bladder neck 14, posterolaterally 20). Eleven patients (18.3%) had biochemical progression. 5-year biochemical free progression was 54.5, 76.9 and 87.9% for apex, bladder and the posterolateral location, respectively (p < 0.05). CONCLUSIONS: In the present study, a positive surgical margin at the apex was associated with worse clinical prognosis compared to the bladder neck and posterolateral locations. PMID- 12740495 TI - Prognostic implication of microvascular invasion in biochemical failure in patients treated with radical prostatectomy. AB - INTRODUCTION: Micrometastasis before radical prostatectomy (RP) is considered to be related to the biochemical failure after surgery in patients with prostate cancer. To predict the biochemical failure, we evaluated microvascular (lymphatic/vascular) invasion and other pathological findings as prognostic factors for biochemical failure after RP. MATERIALS AND METHODS: Eighty-two men who underwent RPs and received neither preoperative nor postoperative adjuvant therapy were analyzed. Pathological findings of each patient were carefully reviewed to determine pathological factors predicting biochemical failure. The influence of pathological findings on the biochemical failure was evaluated. RESULTS: The overall biochemically assessed disease-free survival rate was 55.7% at 3 years. By univariate analysis, Gleason sum, capsular penetration, and microvascular invasion were significantly associated with the rate of biochemical failure. Microvascular invasion was strongly correlated with Gleason sum, capsular penetration, surgical margin, perineural invasion, and pathological stage. By multivariate analysis, Gleason sum, capsular penetration, and microvascular invasion were independent predictors of biochemical failure. CONCLUSION: This study suggests that in addition to Gleason grade and capsular penetration, microvascular invasion is one of independent prognostic factors in patients with prostate cancer treated with RPs. PMID- 12740496 TI - Prostate lymphoscintigraphy and radio-guided surgery for sentinel lymph node identification in prostate cancer. Technique and results of the first 350 cases. AB - INTRODUCTION: Having in mind the promising results of lymphoscintigraphy and intraoperative gamma probe application for the detection of sentinel lymph nodes (SLN) in malignant melanoma, breast and penis cancer, we tried to identify the SLN in prostate cancer by applying a comparable technique. MATERIALS AND METHOD: 350 patients with prostate cancer were examined after providing informed consent. The day before pelvic lymphadenectomy technetium-99m nanocolloid was transrectally injected into the prostate under ultrasound guidance. A single central application was done per prostate lobe in most cases. Activity attained 90- 400 MBq, and the total injected volume was about 2-3 ml. Hereafter, lymphoscintigraphy was carried out. Those lymph nodes having been identified as SLN by means of gamma probe detection and lymphoscintigraphy were removed intraoperatively. Later, most of the cases had different types of pelvic lymphadenectomy. SLN received serial sections and immunohistochemistry, non-SLN step sections. RESULTS: 335 patients showed at least 1 SLN in lymphoscintigraphy. 24.7% had lymph node metastases. In 2 patients, metastases in non-SLN were found without at least one SLN being affected (false-negative patient). CONCLUSION: Our experience suggests that the SLN identification is not only feasible in breast cancer and malignant melanoma, but also in prostate cancer with a comparable technique. PMID- 12740497 TI - Detecting metastatic pelvic lymph nodes by 18F-2-deoxyglucose positron emission tomography in patients with prostate-specific antigen relapse after treatment for localized prostate cancer. AB - AIM: To evaluate whether positron emission tomography (PET) with (18)F-2 deoxyglucose (FDG) can detect pelvic lymph node metastases in prostate cancer patients who had elevated serum prostate-specific antigen (PSA) levels after treatment. METHODS: Twenty-four patients with a rising serum PSA level after treatment for localized prostate cancer were examined with FDG-PET before pelvic lymph node dissection. All patients had negative findings on whole body bone scan and equivocal pelvic computed tomography (CT) results. The results of FDG-PET were then compared to the histology of the pelvic lymph nodes obtained at surgery. RESULTS: Lymph node metastases were detected by histopathological examination in 16/24 (66.7%) patients. At the sites with histopathologically proven metastases, increased FDG uptake was found in 12/16 (75.0%) patients. In addition, there were 4 patients with false-negative results, but no patient with a false-positive result on FDG-PET images. The sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of FDG-PET in detecting metastatic pelvic lymph nodes were 75.0, 100.0, 83.3, 100.0, and 67.7%, respectively. CONCLUSIONS: These results suggest that FDG-PET may be a valuable diagnostic tool in the staging of pelvic lymph nodes in patients with PSA relapse after treatment of localized prostate cancer when the whole body bone scan is negative and pelvic CT findings are equivocal. PMID- 12740498 TI - 3-month formulation of goserelin acetate ('Zoladex' 10.8-mg depot) in advanced prostate cancer: results from an Italian, open, multicenter trial. AB - OBJECTIVES: To determine the endocrine effects, efficacy and tolerability of the 3-month formulation of goserelin acetate ('Zoladex' 10.8-mg depot; 'Zoladex' is a trade mark of the AstraZeneca group of companies) in the treatment of patients with advanced prostate cancer. METHODS: Between February 1996 and October 1997, this open, multicentre study enrolled 120 patients with locally advanced (T3/4) or metastatic (N+ or M1) disease, or an increase in prostate-specific antigen (PSA) level after radical prostatectomy. Patients received goserelin acetate 10.8 mg depot every 12 weeks until clinical progression or interruption for adverse events or other reasons. RESULTS: The mean testosterone concentrations were suppressed to the castration range (< or =2 nmol/l) after 4 weeks of treatment and remained suppressed throughout the study. In total, 99/115 (86%) patients had a serum PSA response, and the mean PSA value decreased significantly during treatment (p = 0.006). The mean PSA level at baseline was significantly lower in patients without disease progression compared to those who experienced disease progression (p = 0.0002). Goserelin acetate 10.8-mg depot was well tolerated and there were no injection site reactions. CONCLUSIONS: The goserelin acetate 10.8 mg depot is well tolerated with no injection site reactions. It produces PSA responses and provides reliable suppression of serum testosterone. PMID- 12740499 TI - Primary paratesticular epithelioid leiomyosarcoma. AB - We report a case of a 69-year-old man with a history of left spermatic cord thickening and an enlarging painless mass in the left hemiscrotum. A final diagnosis of paratesticular epithelioid leiomyosarcoma was made. 13 months after treatment he is alive without any signs of tumor recurrence or metastasis. PMID- 12740500 TI - Pneumaturia in a patient with ectopic vas deferens opening in the bladder and agenesis of the ipsilateral seminal vesicle. Case report. AB - Ectopia of the vas deferens (EVD) combined with agenesis of the seminal vesicle, is a rare congenital abnormality. We describe a case of EVD with agenesis of the ipsilateral seminal vesicle, presenting with pneumaturia and frequent urinary tract infection. PMID- 12740501 TI - Recurrent eosinophilic cystitis with peripheral eosinophilia and hyperimmunoglobulinemia E. AB - A case of recurrent eosinophilic cystitis with peripheral eosinophilia and hyperimmunoglobulinemia E, which responded successfully to initial and secondary steroid therapies is reported. PMID- 12740502 TI - Urethral catheter: a pain in the neck! AB - Long-term urinary catheterization is well recognized in the literature as being associated with significant morbidity and mortality. We present a rare and previously unreported complication of a cervical spine abscess resulting from methicillin-resistant Staphylococcus aureus septicaemia originating from the urinary tract in a patient with a urinary catheter. PMID- 12740503 TI - Angiomyolipoma with a caval thrombus. AB - We report the case of a female patient presenting with flank pain. Abdominal ultrasound revealed a tumor of 8 cm in diameter. After abdominal computerized tomography, the tumor was classified as angiomyolipoma with a tumor thrombus in the inferior vena cava. After nephrectomy, the diagnosis was confirmed histologically. To our knowledge, this is the 11th case of a renal angiomyolipoma extending into the vena cava. PMID- 12740504 TI - Spontaneous communication between a simple renal cyst and the pyelocaliceal system with a gas-producing infection. AB - We present the extremely rare case of a 44-year-old woman who presented with right flank pain and high fever, which proved to be a case of spontaneous communication between a renal cyst and the pyelocaliceal system caused by increased pressure in the renal pelvic cavity exerted by a stone leading to infection. PMID- 12740505 TI - A rare case of retrocaval ureter associated with persistent left vena cava. AB - We report a rare case of a retrocaval ureter associated with a left inferior vena cava transposition and with concomitant nephrolithiasis in a young woman already surgically treated during her first years of life for cardiovascular disease. Diagnosis and surgical procedure are described, including the use of a flexible ureterorenoscope to facilitate kidney stone removal. PMID- 12740506 TI - Laboratory tests in the diagnosis of pulmonary embolism. AB - The diagnosis of pulmonary embolism (PE) requires objective testing. However, all imaging techniques have their own limitations and costs and cannot be performed in every patient with suspected PE. After decades of unfruitful research, several laboratory tests have been evaluated for suspected PE, the most promising being the D-dimer test. As a general rule, the specificity of D-dimers is too low to confirm PE. Conversely, several (but not all) D-dimer assays have a high sensitivity for diagnosing PE. Outcome studies indicate that the Vidas D-dimer and SimpliRED D-dimer can be used safely to withdraw anticoagulation when the pretest probability of PE is low (SimpliRED) or when it is low or moderate (Vidas). These results may however not apply to other D-dimer assays and clinicians should know the characteristics of the test used in their hospital. Blood gas analysis does not have sufficient sensitivity and specificity to confirm or exclude PE, but it may be used to evaluate the clinical probability of PE before other testing is done. The diagnostic value of the alveolar dead space fraction in patients with suspected PE is currently investigated. Initial data suggest that it needs to be combined with a D-dimer test to safely exclude PE. Brain natriuretic peptide and cardiac troponin have limited usefulness for diagnosing PE, but both tests may identify patients with a poor prognosis, in whom more aggressive treatment may be warranted. PMID- 12740507 TI - Do CHF patients align with COPD patients during exercise? PMID- 12740508 TI - Of mice and men, or: a pill for emphysema? PMID- 12740510 TI - Lung attenuation measurements in healthy young adults. AB - BACKGROUND: High-resolution computed tomography (HRCT) attenuation measurements may be more sensitive in finding early emphysematous changes in relatively young subjects than lung function measurements. OBJECTIVES: To define lung attenuation parameters in smokers and never-smokers. METHODS: A prospective comparative study in a university hospital setting was designed with 20 healthy smoking and 20 nonsmoking volunteers. Attenuation measurements on spirometrically controlled HRCT at three levels in the upper half of the lungs at 10% and 90% of vital capacity (VC10% and VD90%) were done, and lung function measurements were performed. RESULTS: Mean lung attenuation (MLD) and pixel index (PI) were correlated with lung function and smoking history. Small attenuation differences in the left and right lung were found but no sex-related differences. At main carina (MC) level, the PI was higher at VC90% (p < 0.0001) but lower at VC10% (p < 0.01) compared to the apex. Age correlated with attenuation parameters at VC10%, whereas for pack-years no correlation was found. There were attenuation correlations with VC (PI: R = 0.31, p < 0.05 at VC10%/90%), residual volume (MLD: R = -0.31, p < 0.05 at VC10%), and total lung capacity (PI: R = 0.31, p < 0.05 at VC10%/VC90%). CONCLUSIONS: Lung attenuation during inspiration was lower at the MC level than at the top, but higher on expiratory scans. No sex-related differences were found. Lung attenuation decreases with age on expiratory scans. This seemed to be of more importance than the amount of pack-years of smoking. A relationship with lung function parameters is not uniformly proven. PMID- 12740511 TI - Retrospective evaluation of patient compliance in continuous oxygen therapy. AB - BACKGROUND: Prescription of continuous oxygen therapy (COT) for correct indications, adequate education of the patients, and a regular and sufficient oxygen intake are essential. OBJECTIVE: To evaluate the indications of COT prescriptions in six different centers around Istanbul and patient compliance. METHODS: The initial data of the cases were taken from hospital records and throughout the study personal visits were done as well as arterial blood gas (ABG) analyses. RESULTS: 86 patients out of 110 (78.1%) were found to be eligible candidates for COT. Daily duration of therapy was more than 15 h in 55 (43.3%) and more than 12 h in 14 (11%) of the cases. During therapy, mean PaO(2) values have increased from 52.1 +/- 11 to 58.6 +/- 12 mm Hg (p < 0.001), while mean PaCO(2) values have decreased from 50.16 +/- 9.6 to 48.02 +/- 9 mm Hg (p < 0.05). 98 of the cases (77.1%) stated that they had benefited from the therapy. CONCLUSIONS: We conclude that COT is an effective treatment modality, but patient compliance remains a problem. PMID- 12740509 TI - Airflow limitation and breathing strategy in congestive heart failure patients during exercise. AB - BACKGROUND: Congestive heart failure (CHF) patients experience dyspnea on exertion and therefore have decreased exercise tolerance. OBJECTIVE: This study explores the hypothesis that stable New York Heart Association (NYHA) class III CHF patients without a history of pulmonary disease exhibit airflow limitation with increasing exercise. METHODS: We characterized flow limitations and breathing reserves at baseline, during exercise before anaerobic threshold (pre AT), and after anaerobic threshold (post-AT) in CHF patients and normal subjects. Data were collected in the form of maximal flow volume loops and subsequent tidal flow volume loops at baseline and during exercise. Expiratory flow limitation was expressed as percent of tidal volume that corresponded with overlap of the tidal flow volume loops and maximal flow volume loops during expiration. The area directly between the maximum flow volume loops and the tidal flow volume loops during the expiratory phase is expressed as expiratory flow volume reserve (EFVR). RESULTS: CHF patients experienced expiratory flow limitation during exercise (pre-AT and post-AT) that was significantly increased compared to baseline and to normal subjects at similar exercise levels (CHF, baseline 8.5 +/- 7, pre-AT 37 +/- 10, post-AT 38 +/- 8%, n = 9, p < 0.05). Both CHF patients and normal subjects increased EFVR during exercise, but only the normal subjects increased EFVR to a significantly different value at post-AT exercise levels (normal subjects, 9.5 +/- 2, 11 +/- 2, 32 +/- 4%, n = 7, p < 0.05). Both CHF patients and normal subjects increased end inspiratory lung volume (EILV) during exercise, but only the normal subjects significantly increased EILV at post-AT exercise levels (normal subjects, 49 +/- 4, 55 +/- 5, 76 +/- 4%, p < 0.05). Inspiratory capacity (IC)/forced vital capacity (FVC) ratios were increased in CHF patients compared to normal subjects. However, IC/FVC values did not change during exercise in either group. CONCLUSIONS: CHF patients cannot utilize their full respiratory capacity during exercise secondary to expiratory flow limitation and an inability to increase EILV and EFVR. PMID- 12740512 TI - Increased vitronectin and endothelin-1 in the breath condensate of patients with fibrosing lung disease. AB - BACKGROUND: Non-specific interstitial pneumonia (NSIP) and fibrosing alveolitis associated with systemic sclerosis (FASSc) are diseases of unknown aetiology that are characterised by the accumulation of mononuclear cells, followed by the progressive deposition of collagen within the interstitium and subsequent destruction of lung airspace. Better understanding of mediators involved in fibrosis may be useful for early diagnosis and in clinical monitoring of disease progression. OBJECTIVE: The aim of this study was to investigate the presence of two profibrotic markers, the vitronectin and the endothelin-1 (ET-1) in the airways of NSIP and FASSc patients. METHODS: Ten NSIP (6 males, age 57 +/- 2 years) and 15 FASSc (8 males, age 55 +/- 4 years) patients were recruited along with 10 normal subjects (4 male, age 52 +/- 2 years). Vitronectin and ET-1 concentrations were measured in their breath condensate, using a specific enzyme immunoassay. RESULTS: Higher levels of vitronectin and ET-1 were observed in NSIP and FASSc patients [median 92.8 (91.7-93.9) microg/ml; median 8.3 (7.9-9.3) pg/ml] than in control subjects [median 80.3 (89.3-91.4) microg/ml; p < 0.01; median 5.3 (4.9-5.9) pg/ml, p < 0.0001]. We also found increased concentrations of vitronectin in patients with clinical deterioration compared to those remaining stable and in ex-smokers compared to non-smokers and, increased vitronectin and ET-1 in patients treated with steroids compared to untreated patients. CONCLUSION: These findings justify further studies of vitronectin and ET-1 levels in exhaled breath condensate, as a means of monitoring activity and predicting progression of pulmonary fibrosis. PMID- 12740513 TI - Serum concentration of D-dimers as a marker of the activation of the fibrinolytic system in patients undergoing bronchoscopic and thoracoscopic investigations. AB - BACKGROUND: Hypercoagulability is common even after minimally invasive surgical techniques and is pathogenetically linked to postoperative thrombotic and cardiac complications. The activation of the coagulation and fibrinolytic systems after bronchoscopic and thoracoscopic investigations has not yet been elucidated. OBJECTIVE: The aim of our study was to evaluate whether bronchoscopic and thoracoscopic investigations activate the fibrinolytic system. METHODS: This study assessed cross-linked fibrinogen degradation products (D-dimer) in 120 patients after bronchoscopy with and without biopsies and/or bronchoalveolar lavage and after thoracoscopy with biopsies of the parietal pleura. RESULTS: Both bronchoscopy and thoracoscopy induced a minor but significant increase in D dimers in most patients, reversible mostly within 24 h. In rare cases, distinct increases lasting at least 24 h occurred. CONCLUSIONS: The slight and temporary increase in serum D-dimers induced by bronchoscopy and thoracoscopy is nonsignificant in most patients; however, in some cases, the coagulation/fibrinolytic systems may become markedly activated. PMID- 12740514 TI - Platinum-based, leukocyte-depleting chemotherapy does not alter induced sputum markers of neutrophilic inflammation in COPD patients with unresectable non-small cell lung cancer. AB - BACKGROUND: Neutrophilic inflammation is a major feature of chronic obstructive pulmonary disease (COPD), and several novel therapies aim at the suppression of neutrophils in COPD. Due to the abundance and redundancy of mediators involved in neutrophilic inflammation, there is an ongoing controversy about the feasibility of such anti-neutrophilic approaches. Systemic chemotherapy has broad side effects, including neutrophil toxicity. OBJECTIVES: In this observational study, we have measured cellular and neutrophil-related inflammatory markers in induced sputum of COPD patients with unresectable non-small cell lung cancer (NSCLC) undergoing platinum-based chemotherapy. METHODS: 15 COPD/NSCLC patients were followed during their first course of chemotherapy with cisplatin (60 mg/m(2) days 1 and 7) and etoposide (100 mg/m(2) days 3, 4, and 5). Sputum induction was performed before, and 3 weeks after chemotherapy. Peripheral blood count, sputum total cells and differentials, and the concentrations of the inflammatory markers interleukin (IL)-8, and matrix metalloproteinase (MMP)-9 in sputum supernatant were analyzed. RESULTS: Similar to COPD controls (n = 12), COPD/NSCLC patients had increased levels of absolute and relative sputum neutrophils, IL-8, and MMP-9 at baseline, when compared with healthy controls (n = 14, p < 0.001, all comparisons). After chemotherapy, there was a significant reduction in peripheral blood leukocytes (pre: 10,736 +/- 550, post: 6,536 +/- 1,064 cells/microl, p = 0.002) and log neutrophils (pre: 8.9 +/- 0.09, post: 8.1 +/- 0.2 cells/microl, p = 0.004), whereas log sputum neutrophils (pre: 0.3 +/- 0.37, post: 0.18 +/- 0.3 cells x 10(6)/ml, p = 0.1), IL-8 (pre 15.9 +/- 3.8, post: 17.7 +/- 3.6 ng/ml, p = 0.7), and log MMP-9 (pre: 5.3 +/- 0.57, post: 5.6 +/- 0.7 ng/ml, p = 0.33) remained unchanged. CONCLUSION: A single course of platinum-based chemotherapy markedly decreases peripheral blood neutrophils, but has no effect on inflammatory patterns of induced sputum in COPD patients with unresectable NSCLC. PMID- 12740515 TI - The effects of shallow versus deep endotracheal suctioning on the cytological components of respiratory aspirates in high-risk infants. AB - BACKGROUND: The harmful effects of mechanical ventilation and suctioning are compounded if endotracheal suctioning (ETS) is inappropriately performed. Deep ETS involves catheter insertion into the endotracheal tube until resistance is met. Shallow ETS may be beneficial in lessening mechanical irritation to the first bronchial layers. However, clinical observation reveals wide variation in the length of the suction catheter for ETS in high-risk infants. OBJECTIVE: The study was conducted to examine the effects of deep and shallow ETS on the cytological components of respiratory aspirates from high-risk infants. METHODS: A cross-over experimental study was performed in 22 high-risk infants with a mean birth weight of 2200 g. Whether deep or shallow ETS was conducted first was determined randomly. The numbers of (1). columnar cells [CC - ciliated (CCC) and nonciliated (NCC)], (2). fresh clustered columnar cells (CLCC), and (3). Curschmann's spirals (CS), a mucus cast residing inside the lower terminal airways, in the respiratory aspirates were compared between the two ETS protocols. RESULTS: No statistical differences were found in the quantities of CC, CLCC and NCC between shallow and deep ETS. However, greater quantities of CLCC were observed in the deep ETS aspirates than in the shallow ETS aspirates. CONCLUSIONS: Detachment of larger amounts of clustered columnar cells from the respiratory epithelium without the guarantee that lung secretions from the lower airways will be removed questions the justification of deep ETS in high-risk infants. PMID- 12740516 TI - Endobronchial ultrasound. AB - Complex technical problems interfered with the application of thoracic ultrasound (US) for studies and clinical research. Moreover, in contrast to radiologists, cardiologists, gastroenterologists, internists, obstetricians, gynecologists and others, pulmonologists were not trained in the basics of US images. However, endoscopic US methods were developed in the last 20 years and these methods also provided important results for pulmonologists. As soon as the technical problems interfering with US application in air-containing spaces were solved, endobronchial US (EBUS) became a valuable technique as well. With EBUS, the delicate multilayer structure of the tracheobronchial wall can be analyzed. This knowledge became decisive for the management of early cancer in the central airways. These lesions can undergo local treatment instead of surgical intervention if the bronchial cartilage is intact and if the adjacent lymph nodes are not involved. EBUS proved valuable as well for the staging of more advanced lung cancer, especially with regard to endoluminal, intramural and extraluminal tumor spread. Endobronchial endosonographers are able to diagnose mediastinal lymph nodes similar to the experience of gastrointestinal endosonographers. EBUS guided transbronchial needle aspiration (TBNA) improved the results of N-staging of lung cancer, especially in difficult lymph node levels without any clear endoscopic landmarks. The possibility of identifying N2 and N3 stages by means of a nonsurgical procedure can modify the management of lung cancer and decrease the number of unnecessary surgical interventions. EBUS can reduce the need for more invasive procedures such as thoracoscopy or mediastinoscopy. It is also useful for biopsying peripheral lesions or solitary pulmonary nodules instead of fluoroscopic guidance and also plays an important role in the strategy of interventional endoscopy. PMID- 12740517 TI - Evaluation of virtual reality bronchoscopy as a learning and assessment tool. AB - BACKGROUND: Conventional training in bronchoscopy involves a trainee performing on a real patient under supervision. This method of training is not only expensive, but there is also potential for increased patient discomfort. Simulators permit the acquisition of necessary technical skills required for the procedure. Virtual reality (VR) has been an integral part of training in aviation, and the application of this technology in medical training needs to be evaluated. OBJECTIVE: This study was conducted to evaluate the efficacy of a VR bronchoscopy simulator as a learning and assessment tool. METHODS: The bronchoscopic simulator (HT Medical Systems, Maryland, USA) is a VR computer programme. The simulator has the ability to assess competence by a set of parameters, which formed the data for the study. Nine novices without previous bronchoscopic experience formed the study group (group 1). Nine experienced bronchoscopists having performed between 200 and 1000 bronchoscopies formed the other group (group 2). We assessed the efficacy of the system as a learning tool by studying whether there was a significant difference between the first and subsequent sessions of the subjects from group 1 and by comparing the performance of the two groups. Statistical analysis was done using the Mann-Whitney U test and the Wilcoxon signed ranks test. RESULTS: There was a significant difference in performance between the first attempt of group 1 and the performance of the experts in terms of percentage of segments visualised and number of wall collisions and the economy of performance. Among the subjects from group 1, there was a significant improvement in percentage of segments visualised by the third attempt (p = 0.04), in the economy of performance by the sixth attempt (0.008) and in the number of wall collisions by the sixth attempt (0.024). When each attempt of the novices was compared with the performance of group 2, the significance in the difference of the percentage of segments studied (p = 0.09) and the economy of performance disappeared by the third attempt (0.06), while the difference in the number of wall collisions disappeared by the fifth attempt (p = 0.06). CONCLUSIONS: This study has been able to establish the face, construct and content validity of the simulator and the potential for it to be an effective training tool. PMID- 12740518 TI - Retinoic acid does not affect alveolar septation in adult FVB mice with elastase induced emphysema. AB - BACKGROUND: Administration of ALL-TRANS retinoic acid (ATRA) to adult Sprague Dawley rats with emphysema induced by porcine pancreatic elastase (PPE) reversed the emphysema perhaps by inducing new alveolar formation. OBJECTIVE: A study was conducted to determine whether ATRA can induce new alveolar septa and reverse the airspace enlargement caused in adult mice by PPE treatment. METHODS: 48 FVB mice were divided into 6 groups. Three groups received 15 microg of PPE in 0.1 ml of 0.9% saline and 3 groups received 0.1 ml of saline, intratracheally. Starting at day 22, the mice received 12 daily intraperitoneal injections of cottonseed oil, with or without ATRA (12.5 microg or 50 microg). The mice were killed for study 1 day after the last injection. RESULTS: Measurements of plasma and lung tissue ATRA levels showed statistically significant elevated levels after the 50-microg but not after the 12.5-microg doses of ATRA. In situ hybridization studies of elastin and alpha(1)(I) collagen mRNA expression in pulmonary parenchyma as well as in airways and blood vessels showed no effect of ATRA. Airspace size was determined by the mean linear intercept (Lm) method. The Lm of the groups receiving PPE and ATRA (46.2 +/- 4.1 microm, mean +/- SD) was not significantly different from the group receiving PPE and oil (47.8 +/- 6.0 microm). The Lm for groups receiving saline and ATRA (40.6 +/- 2.5 microm) were not significantly different from the group receiving saline and oil (41.0 +/- 2.7 microm). Comparison of the fixed lung volume data and calculated internal surface area also showed no differences between the control and ATRA-treated groups. CONCLUSION: ATRA treatment does not affect airspace size or expression of elastin or alpha(1)(I) collagen mRNA in adult FVB mice with PPE-induced emphysema. PMID- 12740519 TI - Black bronchoscopy. PMID- 12740520 TI - Orthodeoxia without platypnea from interatrial defect associated with persistent left superior vena cava in the absence of pulmonary hypertension. AB - Orthodeoxia-platypnea is a rare but increasingly recognized syndrome of upright hypoxemia usually associated to breathlessness relieved by recumbency. We report the case of isolated orthodeoxia discovered in a military recruit who referred only fatigability as the clinical symptom after a forced march. Transesophageal echocardiography demonstrated the presence of interatrial defect ostium secundum type with the persistence of left superior vena cava draining into coronary sinus. Right-to-left shunt was visualized by peripheral saline contrast infusion, despite normal right hemodynamics at heart catheterization. Hypoxemia recovered after the percutaneous closure of the interatrial defect. Orthodeoxia and platypnea could occur as separate disease manifestations, the latter probably being a rarer acute event, whereas orthodeoxia was underestimated and potentially earlier screenable. PMID- 12740521 TI - Spontaneous pneumomediastinum and hemopneumothoraces secondary to cystic lung metastasis. AB - We report a case of a cystic metastasis to the lung from an angiosarcoma of the scalp in a 75-year-old man who complained of hemoptysis. A chest CT scan showed multiple thin-walled pulmonary cysts, bilateral pneumothoraces, small nodules and pneumomediastinum. Histologic examination revealed pleural infiltration of angiosarcoma cells. One month later, a high-resolution CT scan showed that the cysts had rapidly developed into large lesions. PMID- 12740522 TI - Bilateral and unilateral spontaneous massive hemothorax as a presenting manifestation of rare tumors. AB - Spontaneous true hemothorax is quite a rare manifestation of a presenting disease. This is a report of two patients, one with bilateral spontaneous massive hemothorax as a presenting manifestation of angiosarcoma involving the lungs and pleura, and the other with unilateral spontaneous hemothorax and hemorrhagic shock as a presenting manifestation of 'cystic' chondroblastoma. Differential diagnosis of spontaneous true hemothorax and its evaluation and management are discussed. PMID- 12740523 TI - Cervico-mediastinal lymphadenopathy as a paradoxical response to chemotherapy in pulmonary tuberculosis. A case report. AB - We report a case of pulmonary tuberculosis who developed tuberculous mediastinal and later cervical lymphadenopathy while on antituberculous chemotherapy. She responded to the addition of oral corticosteroids to her regimen. The relevant literature regarding the pathogenesis, varied manifestations and the clinical importance of recognising a paradoxical response is discussed. PMID- 12740524 TI - Gardening in greenhouses as a risk factor for silicosis. AB - Silicosis is a typical occupational disease, although some cases caused by non industrial exposure have also been reported. We saw a 53-year-old male gardener with recurrent non-productive cough. A routine radiograph of the chest showed bilateral pulmonary nodules and subsequent computed tomography suggested that the infiltrates could be metastases. Open lung biopsy revealed nodules consisting of fibrotic tissue while the presence of birefringent silica particles was observed by polarised light microscopy. Mineralogical analysis of the substrata from the patient's workplace revealed an SiO(2) concentration of 31%. This case indicates that the inhalation of siliceous particles in a closed environment such as a greenhouse is a risk factor for silicosis. PMID- 12740525 TI - Complement in different stages of HIV infection and pathogenesis. AB - The complement system is one of the most important weapons of innate immunity and is involved in all infectious processes. It is not only a mechanism for direct protection against an invading pathogen but it also interacts with the adaptive immunity to optimize the pathogen-specific humoral and cellular defence cascade in the body. One of the greatest challenges for the complement system is infection by HIV with its chronic course and sequential destruction of immune cells and immune organs. Due to its dual role as direct effector and as fine tuner of adaptive immunity, we focussed on complement in this review and analysed in detail the contribution of complement to the antiviral defence and to HIV pathogenesis on the one hand and the complement evasion strategies of the virus on the other hand. In the present review, this interplay between complement and the virus is illuminated for the three different stages of HIV pathogenesis and for events during therapy: (1) the acute infection process with the early events in mucosa and serum; (2) the asymptomatic stage with the complex interplay between complement-induced lysis and viral evasion strategies; (3) the symptomatic infection and AIDS stage with progressive destruction of the lymph nodes, opportunistic infections and development of neuropathogenesis, and (4) finally, during highly active antiretroviral therapy and in vaccination approaches. PMID- 12740526 TI - Cloning and characterisation of two IgE-binding proteins, homologous to tropomyosin and alpha-tubulin, from the mite Lepidoglyphus destructor. AB - BACKGROUND: The dust mite Lepidoglyphus destructor is a major source of mite allergy in European rural environments, but it also causes allergy in urban populations around the world. We have previously cloned, sequenced and expressed several allergens from L. destructor (Lep d 2, Lep d 5, Lep d 7 and Lep d 13). The aim of this study was to identify and clone additional allergens from L. destructor, and to evaluate their IgE-binding reactivities. METHODS: PCR and screening with sera from L. destructor-sensitised individuals were used to isolate new clones from a phage display L. destructor cDNA library. The complete coding sequences of the clones were determined and expressed as His(6)-tagged recombinant proteins in Escherichia coli. The recombinant proteins were analysed by SDS-PAGE, immunoblotting and mass spectrometry. RESULTS: Two new clones, showing homology to tropomyosin and alpha-tubulin in several species, were isolated from the phage display L. destructor cDNA library. Due to its homology to group 10 dust mite allergens, the tropomyosin clone was named Lep d 10. The IgE-binding frequencies of the recombinant Lep d 10 and alpha-tubulin were 13% (18/136) and 12% (11/95), respectively, among subjects with IgE reactivity to mites and/or crustaceans. CONCLUSIONS: Two new allergens from L. destructor have been identified and can now be added to the repertoire of recombinant L. destructor allergens. In addition, both these allergens belong to highly conserved protein families and may be important for evaluation of allergenic cross-reactivity. PMID- 12740527 TI - Comparison of allergen-induced late inflammatory reactions in the nose and in the skin in house dust mite-allergic patients with or without asthma. AB - BACKGROUND: It remains to be established which factors contribute to the occurrence of asthma in allergic individuals. We hypothesized that differences in the late allergic inflammatory reaction to allergen between asthmatic and non asthmatic house dust mite-allergic individuals might contribute to the difference in the clinical presentation of allergy. AIM: To compare allergen-induced changes in parameters for cellular inflammation during the phase of the late allergic reaction in the skin and nose, in house dust mite-allergic individuals with or without asthma. MATERIAL AND METHODS: Nasal and dermal allergen challenges with house dust mite (Dermatophagoides pteronyssinus) extract were performed in 52 house dust mite-allergic individuals, of whom 26 had mild to moderate persistent asthma and 26 had perennial rhinitis without current or past asthmatic symptoms. Serial nasal lavage samples were analyzed for the presence of inflammatory cells (eosinophils and neutrophils) and soluble markers associated with cellular inflammation [interleukin-5 (IL-5), interleukin-8 (IL-8), eosinophil cationic protein (ECP) and myeloperoxidase (MPO)]. Macroscopic late phase skin reactions were studied after intracutaneous skin tests with house dust mite extract. RESULTS: Fixed dose nasal allergen provocation elicited a similar degree of immediate allergic reaction as judged by plasma protein exudation and histamine concentrations in asthma and non-asthmatic rhinitis. Subsequently, no differences between groups were found during the phase of the late allergic reaction (4-24 h) in inflammatory cell influx, plasma protein leakage, ECP or MPO. Likewise, there were no differences in levels of chemotactic cytokines IL-5 and IL-8. In agreement with the results of nasal challenge, the late skin reaction after dermal challenge with a fixed allergen dose and after an allergen dose 10,000 times above the skin threshold for an early skin reaction did not differ between the groups. CONCLUSION: House dust mite-allergic patients with or without asthma have very similar late allergic inflammatory reactions in the skin and in the nose after allergen challenge. Hence, it is unlikely that the occurrence of pulmonary symptoms in asthma is explained by a general tendency of asthmatics to have an enhanced late allergic cellular inflammatory response. Nasal and dermal allergen provocations are adequate models to study allergen-induced inflammation but probably lack the pivotal link which is essential for the development of asthma. PMID- 12740528 TI - Possible modes of allergen-specific sensitization and boosting in an atopic child. AB - Several studies document that allergen-specific IgE levels are boosted by allergen contact via the respiratory tract in allergic patients. Only few data are available on whether other routes of allergen contact have an influence on systemic IgE responses. We report the case of a boy who developed egg allergy after heavy consumption of eggs by the mother during pregnancy and breast feeding. In contrast to other children who outgrow egg allergy during the first years of life, the boy experienced further dramatic increases in hen egg-specific IgE antibodies after prolonged consumption of ostrich eggs containing cross reactive allergens. IgE antibodies to most of the important respiratory allergens remained either low or not detectable. The dramatic increases in hen egg-specific IgE antibody levels after oral intake of allergens demonstrate that systemic IgE responses in allergic patients can be strongly boosted by allergen contact via routes other than the respiratory tract. PMID- 12740529 TI - Construction of an sIgE:FLAG-mIgE:GFP reporter mouse strain. AB - Like all other immunoglobulins, IgE can be secreted into the blood or expressed as a membrane receptor on the surface of B lymphocytes. Secreted immunoglobulins trace the antigen and contribute to its destruction. Membrane immunoglobulins accompany the B cell along its differentiation pathway, regulating processes like the induction and maintenance of immunological memory and differentiation of plasma cells. The regulation of the expression of IgE is very complex. A lot of positive and negative regulators influence the synthesis of IgE. In previous publications, we were able to show that the membrane IgE (mIgE) antigen receptor itself controls the quantity and quality of serum IgE produced. However, the knowledge about the regulatory function of the antigen receptor on these processes is at best limited. In the present paper, we present the construction of a reporter mouse strain, which will help us to follow an mIgE-bearing B cell during the immune response more precisely. PMID- 12740530 TI - Involvement of arachidonic acid in nonimmunologic production of superoxide in mast cells. AB - BACKGROUND: A number of different molecules are known to be involved in the signal pathway to release histamine from mast cells, among which arachidonic acid (AA) is one of the key mediators. On the other hand, we found that the application of compound 48/80, a typical histamine liberator, generated superoxide in mast cells. In the present study, we investigated the mechanism of superoxide production in mast cells with respect to AA signaling in conjunction with a fine structural analysis. METHODS: Superoxide production was monitored by chemiluminescence in rat peritoneal mast cells and their subfractions after various treatments. For scanning electron micrography, the conditions for fixation and freeze-fracture were optimized to get natural fine images. RESULTS: Compound 48/80 induced superoxide production in the isolated mast cells and some of their subfractions possibly through intracellular increase in Ca(2+) concentration, activation of cytosolic phospholipase A(2), and release of AA. DISCUSSION: The present results indicate the critical role of AA in the signal pathway to generate superoxide from mast cells in response to compound 48/80. PMID- 12740531 TI - Effect of rapid eye movement sleep deprivation on allergen-induced airway responses in a rat model of asthma. AB - BACKGROUND: Nocturnal airway narrowing is a common characteristic of asthma that remains poorly understood. Sleep itself or rapid eye movement (REM) sleep has been suspected to play a role in the etiology of nocturnal asthma; however, only inconsistent findings have so far been produced. The purpose of this study was to examine the effect of sleep deprivation, particularly REM sleep, using the classical platform (flowerpot) method on asthmatic responses in a rat model of asthma, in a situation of immobilization stress. METHODS: After the rats had been randomly divided into three groups of 9 including (1) a caged control group, (2) a large (sham) platform group, and (3) a small platform group, we investigated the effect of REM sleep deprivation over a 72-hour period on both the antigen induced immediate asthmatic response (IAR) and late asthmatic response (LAR) including the changes of stress hormones. RESULTS: The small platform group showed significantly lower plasma histamine levels and higher plasma adrenaline levels during the IAR compared to the control group (p < 0.05). The number of eosinophils in either the bronchoalveolar lavage fluid or the bronchial lamina propria during the LAR in the small platform group was reliably suppressed compared to the other groups (p < 0.05). No significant differences were found in the plasma corticosterone and noradrenaline levels among the three groups, irrespective of the phase of the asthmatic response. CONCLUSIONS: Our results indicate that REM sleep contributes to nocturnal asthma, possibly due to an alteration of the sympathetic nervous function. PMID- 12740532 TI - Comparative effects of desloratadine versus montelukast on asthma symptoms and use of beta 2-agonists in patients with seasonal allergic rhinitis and asthma. AB - BACKGROUND: Asthma and seasonal allergic rhinitis (SAR) are recognized as manifestations of a single airway disease. Desloratadine has demonstrated efficacy in treating SAR symptoms, including nasal obstruction. METHODS: Safety and efficacy of desloratadine and montelukast each were assessed in a double blind, placebo-controlled trial of patients with SAR and symptoms of asthma, who were assigned randomly to once-daily treatment with desloratadine 5 mg, montelukast 10 mg, or placebo for 4 weeks. Change from baseline of AM/PM reflective total asthma symptom severity scores (TASS), FEV(1), individual asthma symptom scores, and beta(2)-agonist usage were assessed. RESULTS: Desloratadine and montelukast each were associated with statistically significant reductions from baseline in the mean TASS averaged over the 4-week period (p < or =0.022 vs. placebo). Individual asthma symptom scores also improved significantly for both therapies (p < or = 0.05). Patients treated with desloratadine or montelukast demonstrated improvement from baseline in FEV(1) versus placebo; significant improvement was seen in a subset of patients with baseline FEV(1) <80% of predicted normal (both p < 0.05). Both active therapies significantly reduced beta(2)-agonist use (both p < 0.01). Improvements for both therapies were comparable for all efficacy parameters; they were tolerated well with adverse event profiles similar to placebo. CONCLUSIONS: Asthma symptoms and beta(2) agonist were improved significantly in patients with concomitant SAR and asthma treated with desloratadine 5 mg as well as montelukast 10 mg once daily. Both therapies significantly improved FEV(1) in a subset of patients with FEV(1) <80% of predicted normal at entry. Improvements in asthma symptoms were comparable for both active treatment groups. PMID- 12740533 TI - Antibody-dependent killing of meningococci by human neutrophils in serum of late complement component-deficient patients. AB - BACKGROUND: Thirty-one Russian patients with late complement component deficiency (LCCD) who had experienced one to five meningococcal infections were immunized with meningococcal polysaccharide vaccine (A + C + W135 + Y) and were followed for 3-8 years. We investigated the potentially protective killing effect of human neutrophils (PMNL) on serogroup A and W135 meningococci. METHODS: Meningococci were incubated in LCCD vaccine sera in the absence or presence of PMNL, and the number of live bacteria (CFU) was determined by plating onto chocolate agar. RESULTS: When meningococci were incubated in the LCCD sera alone, exponential growth of meningococci occurred despite the presence of meningococcal antibodies. After the addition of PMNL, meningococci were inhibited in their growth or even eliminated. Group A or W135 meningococci were killed effectively by PMNL in 80% of the sera which were collected 1 month to 1 year after vaccination compared to only 40% in the prevaccination LCCD sera (p < 0.05). Three years after vaccination 67% of the LCCD sera were still capable of promoting killing (and even 90% after revaccination). The rate of killing correlated with the concentration of serogroup-specific immunoglobulins. In 83% of the 72 LCCD sera with more than 5 microg/ml anti-group A immunoglobulins the killing of group A meningococci was promoted. By contrast, only 21% of 19 samples with lower specific antibody levels showed a PMNL-mediated meningococcal killing (p < 0.05). The same effect was observed for group W135 meningococci. CONCLUSION: PMNL kill meningococci during incubation in LCCD serum; this effect increases after vaccination and depends on both specific antibody and complement. Protection by vaccination may therefore be caused by an increased killing capacity of PMNL. PMID- 12740534 TI - Role of oxygen free radical scavengers in acute renal failure complicating obstructive jaundice. AB - Obstructive jaundice is associated with high morbidity and mortality. Major complications such as pulmonary dysfunction, renal failure and sepsis are frequently encountered. Recent studies and observations suggest that the free oxygen radicals (FORs) produced in obstructive jaundice may play a significant role in the etiopathogenesis of acute renal failure (ARF). Thirty rats were divided into three groups, as sham, control and treatment groups containing 10 rats each. Laparatomy was performed on each animal in the control and treatment groups and common bile ducts were ligated. Common bile duct was observed but was not ligated for the rats in the sham group. Saline solution injection was begun on the first day of surgical procedure and repeated once a day during the following 5 days. The same procedure was performed with oxygen radical scavenger dimethyl sulfoxide (1.5 mg/kg/day i.p.) instead of saline in the treatment group. The rats were sacrificed on the 7th postoperative day. On the 7th postoperative day, the bilirubin, urea and creatinine levels of the control and treatment groups were significantly higher in comparison with the sham group (p < 0.01). Although there was no statistically significant difference between the bilirubin levels of the control and treatment groups (p > 0.05), the urea and creatinine levels in the treatment group were significantly lower (p < 0.01). On the 7th postoperative day, the erythrocyte superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) levels of the control and treatment groups were significantly lower than those of the sham group (p < 0.01), whereas renal and erythrocyte malondialdehyde (MDA) levels were significantly higher (p < 0.01). Although SOD and GSH-Px levels did not differ significantly between the treatment and control groups (p > 0.05), renal and erythrocyte MDA levels of the treatment group were significantly lower than those of the control group (p < 0.01). The histopathological scores were significantly higher in the control and treatment groups (p < 0.01); there was no significant difference between the control and treatment groups (p > 0.05). FORs seem to play a significant role in the etiopathogenesis of renal failure in obstructive jaundice. Antioxidant treatment may decrease oxidative damage due to FORs and may prevent renal failure. PMID- 12740535 TI - Hepatocyte ploidy in regenerating livers after partial hepatectomy, drug-induced necrosis, and cirrhosis. AB - The hepatocyte ploidy was investigated by flow cytometry in regenerating Sprague Dawley rat livers following either drug-induced acute necrosis (single sublethal doses of D-galactosamine or thioacetamide) or drug-induced chronic cirrhosis (repeated thioacetamide injections for 10-18 weeks) and in regenerating livers following 70% partial hepatectomy and was compared with that of normal hepatocytes. Twenty-four hours after partial hepatectomy, a significant decrease in 2n (1 diploid nucleus) hepatocytes and a significant increase in 8n (1 octoploid nucleus) hepatocytes occurred. In contrast, 24 h following induction of acute hepatic failure by single D-galactosamine or thioacetamide injections, a significant increase in 2n hepatocytes was observed, whereas the proportion of 8n hepatocytes remained unchanged. The liver ploidy returned to basal values within 21 days in all cases. In cirrhotic livers induced by chronic thioacetamide injections, the rate of 2n hepatocytes was about ten times that of the controls having the same age, while 4n (1 tetraploid nucleus) and 8n hepatocytes were one third of controls. The binucleation rate was also significantly decreased. PMID- 12740536 TI - Mesh implants in hernia repair. Inflammatory cell response in a rat model. AB - BACKGROUND: In the reinforcement of the abdominal wall with mesh implants, various complications including hernia recurrence, abdominal pain, seroma formation and infection are discussed to depend on the biocompatibility of the alloplastic prosthesis. Particularly macrophages, T-cells and mast cells have been shown to play a major role in the inflammatory response to biomaterials. To approach biocompatibility of surgical meshes we therefore examined the infiltrate of these cells as well as the proliferation rate in response to different clinically applied materials. MATERIALS AND METHODS: Three mesh materials (polypropylene: PP, Prolene; polyethylene terephthalate: PET, Mersilene, and polypropylene/polyglactin: PP + PG, Vypro) were compared after inlay implantation in a standardized rodent animal model. A suture-closed laparotomy served as control. After 7 and 90 days of implantation, histochemical analysis of the inflammatory response to all biomaterials was performed: macrophages (ED3), T cells (CD3), proliferating cells (PCNA) and mast cells (Giemsa) were investigated. RESULTS: In all groups a persisting T-cell response was observed. Colonization of the interface with macrophages showed a pronounced reduction in the PP + PG-mesh group. Infiltration of mast cells at the tissue graft interface showed a time-dependent decrease in the PET- and PP + PG-mesh groups, whereas in contrast, index of mast cells increased in the PP-mesh group. At both time points, indices of proliferation were highest in the PP-mesh group. CONCLUSION: The present data confirm the development of a biomaterial-dependent chronic inflammatory response to surgical meshes with macrophages as the predominant cell type. Further research on the recruitment of inflammatory cells and in particular on the role of mast cells and their granular products should be encouraged. PMID- 12740538 TI - Effect of fluoridated milk on progression of root surface lesions in vitro under pH cycling conditions. AB - The aim of this study was to assess the effect of milk with 0, 2.5 or 5 ppm F on progression and remineralization of caries-like root surface lesions using a pH cycling model. The root surface lesions were created utilizing a partially saturated lactic acid buffer at pH 4.6. Longitudinal sections were cut through the lesion and analyzed using polarized light microscopy (PLM) and microradiography (MRG). The sections were then coated with an acid resistant varnish, except the outer natural surface that would be exposed to water, milk or fluoridated milk and cycled in a de- and remineralizing system for 2 weeks. The lesions were characterized again by PLM and MRG after treatment. A significant reduction in lesion progression was found by PLM and MRG after treatment with either non-fluoridated or fluoridated milk when compared to the control group. Using quantitative MRG, mineral change and distribution in the lesions were recorded. A possible protective effect of fluoridated milk on root surface caries was supported by a reduction in the progression of the lesions and an increase in the mineral within the lesion. PMID- 12740537 TI - Fluoride, beverages and dental caries in the primary dentition. AB - Knowledge concerning risk factors for primary dentition caries in young children is incomplete. Models are presented for caries development using longitudinally gathered fluoride exposure and dietary intake data in the Iowa Fluoride Study. Primary tooth caries examinations were conducted at age 5. Dietary (beverage) and fluoride exposure data were gathered longitudinally from age 6 weeks through 4 years (n = 291); 23% had decayed or filled surfaces. Logistic regression revealed that beverage components and toothbrushing made unique contributions to caries experience. Water consumption (36-48 months), milk consumption (24-36 months), and fluoridated toothpaste brushings (36-48 months) were negatively associated with caries; sugared beverages and milk (6 weeks to 12 months) were positively associated. Although fluoride exposure is important, sugared beverages contribute substantially to caries risk, while water and milk consumption and frequent toothbrushing early can have protective effects. PMID- 12740539 TI - Effect of the routine professional application of topical fluoride on caries and treatment experience in adolescents of low socio-economic status in the Netherlands. AB - The purpose of this study was to study the effect of professionally applied topical fluoride on oral health status at the population level in adolescents with low socio-economic status. Adolescents attending dental clinics where professional fluoride application was a routine procedure (high-fluoride group) were compared to adolescents from other clinics (low-fluoride group). The study consisted of a questionnaire, a clinical examination and two bitewing radiographs. X-rays showed no statistically significant differences in the number of DS, FS or DFS. Clinically, there were no statistically significant differences in the number of FS or in the number of DFS. The low-fluoride group had a statistically significant higher number of DS than the high-fluoride group. The results indicate that professionally applied fluoride has no effect on caries and treatment experience in a population. These results justify a randomized clinical trial on a population level to study the effectiveness of professionally applied topical fluorides. PMID- 12740540 TI - The effect of pH, temperature and plaque thickness on the hydrolysis of monofluorophosphate in experimental dental plaque. AB - Monofluorophosphate (MFP), an anti-caries agent commonly used in toothpaste, is known to be degraded to fluoride and orthophosphate by bacterial phosphatases in dental plaque. We have examined the effect of pH, temperature, plaque thickness and some ions on this process. Both natural plaque and artificial microcosm plaque incubated with purified MFP at pH 4-10 showed an optimum pH of approximately 8 for hydrolysis. Diffusion and concomitant hydrolysis were examined in an apparatus in which artificial plaque was held between rigid membranes separating two chambers. When MFP diffused through a plaque of 0.51-mm thickness over 4 h it was almost completely hydrolysed at pH 8, but hydrolysis on diffusion decreased as the pH deviated from 8. MFP in toothpaste extract showed a similar pH susceptibility to hydrolysis, according to the inherent pH of the toothpaste. Hydrolysis of MFP in the toothpaste was reduced by no more than 10% when compared with a matched-pH control, suggesting that other toothpaste ingredients had no major influence on hydrolysis. Transport was slower and hydrolysis at pH 6 more complete the thicker the plaque, but hydrolysis was not significantly slower at 23 degrees C than at 37 degrees C. The addition of various potential activating or inhibiting ions at 0.1 and 1.0 mmol/l had small and non-significant effects on hydrolysis. The results suggest that MFP toothpaste should be formulated and used to maximise enzymic hydrolysis of this complex anion, and that plaque pH control is probably the most important factor. PMID- 12740541 TI - Effect of a varnish containing chlorhexidine and thymol (Cervitec) on approximal caries in 13- to 16-year-old schoolchildren in a low caries area. AB - BACKGROUND: Previous studies on the effect of the chlorhexidine and thymol containing varnish (Cervitec((R))) have consistently shown a reduction in STREPTOCOCCUS MUTANS, while the data on the caries-inhibiting effect are conflicting. AIM: To measure the effect of a chlorhexidine- and thymol-containing varnish (Cervitec) on approximal caries in a low caries population. MATERIAL: Eighty-five schoolchildren, 13 years of age, entered the study after being selected on the basis of bitewing radiographs. Children with at least one approximal lesion were included after informed consent. METHODS: The study was carried out as a double-blind, randomized controlled trial according to a split mouth design using Cervitec on one side and a placebo varnish on the other side. All approximal surfaces between molars and premolars were treated every 3rd month. After 2 years new bitewing radiographs were taken. A total of 73 children completed the 2-year study. RESULTS: Neither the difference in number of new lesions between the Cervitec-treated and the placebo-treated side (d: -0.21; 95% CI: -0.57; 0.16) nor the mean difference in number of already existing lesions that progressed (d: -0.01; 95% CI: -0.33; 0.30) were statistically significant (paired t test; p > 0.05). CONCLUSION: The results of our study do not indicate an effect of Cervitec on initiation or progression of approximal caries in a population with a low DMFS. PMID- 12740542 TI - The effect of variable energy input from a novel light source on the photoactivated bactericidal action of toluidine blue O on Streptococcus Mutans. AB - Although the combination of toluidine blue O (TBO) dye and laser light at a wavelength of 633 nm has a bactericidal effect, light from laboratory lasers can only be directed externally at a bacterial colony or suspension. In this study a novel delivery system guided the laser light to an 800-micrometer diameter spherical tip (an isotropic tip) from which light radiated producing a uniform sphere of light within the colony or suspension. The system was highly effective in killing TBO-treated Streptococcus mutans NCTC 10449 in stirred planktonic suspension, killing at least 10(9 )cfu/ml. Antibacterial action increased as the delivered energy dose increased. Energy doses of 1.8 J or more produced 100% kills and log reductions of 8-10 cfu/ml. Neither TBO dye nor light alone had a significant antibacterial effect under the experimental conditions used. The existence of a threshold energy, i.e. a minimum energy required before bactericidal action occurred, could not be demonstrated. PMID- 12740543 TI - Effect of a calcium carbonate-based dentifrice on enamel demineralization in situ. AB - Since the effect of calcium carbonate (CaCO(3)) based dentifrice on enamel demineralization is not clearly established, it was evaluated using the IEDT model described by Zero's group in 1992. This study had a crossover design and 10 volunteers were submitted to 3 treatment groups: a negative control, brushing without dentifrice; an active control, brushing with silica-based dentifrice (SiO(2)/MFP group), and the experimental group, brushing with CaCO(3)-based dentifrice (CaCO(3)/MFP). Both dentifrices contained 1,500 microgram F/g (w/w) as sodium monofluorophosphate (MFP). Enamel surface microhardness was determined in the dental blocks and the percentage change in relation to baseline was calculated. Fluoride uptake in enamel and its concentration in 'test plaque' were determined. The results showed that the dentifrice containing CaCO(3)/MFP was more effective than SiO(2)/MFP in reducing enamel demineralization (p < 0.05). A higher concentration of fluoride ion was found in 'test plaque' treated with CaCO(3)/MFP than in the negative control (p < 0.05). The results suggest that CaCO(3) abrasive may enhance the effect of fluoride present in dentifrice on dental caries control. PMID- 12740544 TI - Influence of number of surfaces and observers on statistical power in a multiobserver ROC radiographic caries detection study. AB - The aim of the study was to evaluate the influence of the number of surfaces (N(SURF)) and the number of observers (N(OBS)) on the statistical power of a study comparing the diagnostic accuracies of radiographic systems used for approximal caries lesion detection. A data set consisting of 338 surfaces examined by 10 independent observers using four radiographic systems was available. The presence of a caries lesion was assessed from a 5-point confidence scale. The true lesion diagnosis was established by histological validation. ROC curve areas (A(z)s) were used to express the diagnostic accuracy of the observers with the radiographic systems. Assuming that the A(z)s were tested by a two-way analysis of variance, we performed a simulation study in order to evaluate how the power of this statistical analysis depended on N(SURF) and N(OBS). As a measure of the statistical power we used the standard error of the difference between the expected A(z)s of two systems. The simulations were made with N(SURF) in the range from 25 to 338 and N(OBS) from 2 to 10. The simulations showed that the power increased as a function of the total number of evaluations per system (N(SURF) x N(OBS)), but how this number was attained in relation to the number of surfaces and observers had only marginal influence on the power. Thus, from a statistical point of view it may be concluded, provided that data are analyzed by a two-way analysis of variance, that study designs for comparing the accuracy of several systems can be composed freely in relation to the number of surfaces and observers as long as the total number of evaluations per system are identical. PMID- 12740545 TI - Cultivatable bacteria in dentine after caries excavation using rose-bur or carisolv. AB - To measure the amount of viable bacteria after excavation using conventional rose bur or the chemo-mechanical Carisolv method, a total of 22 lesions were analyzed (one vital tooth per patient) in this open, controlled and randomized study. Two samples per lesion were taken under aseptic conditions using a rose-bur, one superficially in the caries lesion and one after completed excavation. In in vitro tests more material was collected from the hard caries free dentine as compared to the carious dentine. The samples were incubated on blood agar (aerobically and anaerobically), Rogosa (SL) agar and mitis salivarius (MS) agar. For blood agar (aerobic) both methods resulted in a significant decrease in CFU, for blood agar (anaerobic) and MS agar only the Carisolv method resulted in a significant decrease in CFU and for SL agar neither method resulted in a significant decrease in CFU. Comparing CFU before and after excavation, a considerable reduction of CFU was seen ranging from 10(1) to 10(4). Comparing the excavation methods, there were no significant differences, except in the case of blood agar (aerobic), which showed that Carisolv excavation was more effective in reducing CFU. This study indicated that bacterial sampling collected more material from hard dentine as compared from soft. Remaining bacteria after excavation were low in both groups. The Carisolv method seemed to remove bacteria at least up to and possibly beyond the extent of conventional drilling. PMID- 12740546 TI - A new selective medium for Streptococcus mutans and the distribution of S. mutans and S. sobrinus and their serotypes in dental plaque. AB - A new selective medium (MS-MUT) was developed for the isolation of Streptococcus mutans from clinical specimens. The average growth recovery of S. mutans on MS MUT medium was 72.4% of that on MS medium. Growth of Streptococcus sobrinus was significantly inhibited on the medium with an average recovery of 0.034%. In 103 subjects, S. MUTANS was detected at 58.3, 75.0 and 95.7% in the dental plaque of caries-free (CF), caries-inactive (CI) and caries-active (CA) subjects, respectively. S. sobrinus was detected in 8.3, 13.6 and 38.3% of CF, CI and CA subjects, respectively. S. sobrinus alone was detected in only 4.3% of CA subjects. The subjects in whom neither S. mutans nor S. sobrinus were detected were 41.6% in CF and 25.0% in CI. The most predominant serotype was C with a 67% detection rate. S. sobrinus, serotypes D or G were usually found together with S. mutans. PMID- 12740547 TI - Preliminary in vitro assessment of erosive potential using the ultra-micro indentation system. AB - The aim of this study was to measure the change in hardness and modulus of elasticity of enamel and dentine in primary teeth using the ultra-micro indentation system (UMIS) after exposure to potentially erosive beverages and to relate the changes to the chemical composition of the test beverages. Primary molar teeth were sectioned and polished. Indentations were made in enamel and dentine prior to and then after exposure to one of four beverages: distilled water (control), orange juice, Orange Cordial, Fanta((R)) and a 'toothfriendly' Tropical Orange Cordial (Ribena((R))). Each beverage was then analysed. Fanta and Orange Cordial were the only test beverages to show a statistically significant reduction in the enamel hardness in comparison to the control and only Fanta showed a significant reduction in dentine hardness. None of the test beverages significantly reduced the modulus of elasticity of either enamel or dentine. Statistically the pH, phosphate and fluoride concentrations of the beverages were correlated with softening of enamel. Calcium concentration and titratable acidity were correlated with softening of dentine. The UMIS offers another method of measuring the basic mechanical properties of enamel and dentine. This is useful in predicting the erosive potential of substrates. It is likely that the pH, titratable acidity, fluoride, phosphate and calcium content of beverages are all important in determining the potential erosiveness of beverages. PMID- 12740548 TI - Protective influence of experimentally formed salivary pellicle on enamel erosion. An in vitro study. AB - The purpose of this study was to evaluate dental erosion in 0.1 and 1.0% citric acid in vitro by several different methods and to assess the protective potential of experimentally formed salivary pellicle (24 h in vitro). Bovine enamel slabs were embedded in epoxy resin and polished. Erosion was performed in citric acid for 1, 5 or 10 min and recorded as microhardness loss, as changes of surface roughness (R(a), R(t) and R(zDIN)) and as calcium release. Additionally, erosive alterations were observed with scanning electron microscopy. Significant microhardness loss on non-pellicle-covered specimens was measured after 1-min exposure to 0.1% citric acid. Microhardness loss was time- and concentration dependent. Salivary pellicle significantly inhibited both microhardness loss, except after 10-min immersion in 1.0% citric acid, and significantly reduced the increase of surface roughness. There were, however, no significant differences in calcium release between pellicle-covered and non-covered enamel. The results support the general conclusion that salivary pellicle effectively protects enamel surface against short-term erosion in organic acids. PMID- 12740550 TI - Incidence, risk factors and prognosis of stroke and TIA: the need for high quality, large-scale epidemiological studies and meta-analyses. AB - Stroke is a considerable clinical, social and economic burden. In recent clinical trials, a number of strategies have been shown to reduce the risk of stroke and transient ischaemic attack (TIA) in both primary and secondary prevention settings. Whether these treatments are leading to a significant reduction in the incidence of first and recurrent stroke in the clinic, however, remains unclear due to a paucity of high-quality epidemiological data. A similar lack of reliable epidemiological studies has undermined our understanding of the relationship between many potentially important vascular risk factors and stroke risk. Improvement in our knowledge of stroke epidemiology is a prerequisite for the planning of stroke services, the effective application of current stroke prevention strategies, the development of new strategies, and our understanding of the mechanisms of stroke. Future studies must take into account the clinical and pathological heterogeneity of TIA and stroke, and must be powered to allow subtype differences in risk factor relationships and prognosis to be determined reliably. In many cases, this will require meta-analysis of detailed individual patient data from multiple independent studies. PMID- 12740551 TI - Atheroprotective effects of long-acting dihydropyridine-type calcium channel blockers: evidence from clinical trials and basic scientific research. AB - Atherosclerosis is a systemic disease that can ultimately lead to ischaemia and infarction in the heart, brain and peripheral vasculature. According to the "response to injury" hypothesis, endothelial dysfunction is the early event that allows penetration of lipids and inflammatory cells into the arterial wall, contributing to the development of the atherosclerotic lesion. Endothelial dysfunction is causally related to a variety of risk factors for atherosclerosis, including hyperlipidaemia and hypertension. Agents that restore endothelial function and NO bioavailability have beneficial anti-atherogenic activities and can improve cardiovascular outcomes; this has been observed with angiotensin converting enzyme (ACE) inhibitors, statins and certain dihydropyridine-type calcium channel blockers (CCBs). In the Prospective Randomised Evaluation of the Vascular Effects of Norvasc Trial (PREVENT), the CCB amlodipine provided significant clinical benefits compared with placebo, including a marked reduction in cardiovascular morbidity and a reduction in the progression of carotid atherosclerosis. As these beneficial effects of amlodipine have not been observed with other dihydropyridine-type CCBs, it has been proposed that this agent has distinct anti-atherosclerotic properties related to its strong lipophilicity and membrane location. Experimental support for this hypothesis has been obtained from various in vitro and in vivo models of atherosclerosis. These findings support a broader therapeutic role for third-generation dihydropyridine-type CCBs in the treatment of atherosclerosis. PMID- 12740552 TI - Clinical trials: Evidence and unanswered questions--hypertension. AB - Since the pioneering publications of the Hypertension Detection and Follow-up Program (HDFP) and the Multiple Risk Factor Intervention Trial (MRFIT) in the late 1970s and early 1980s, it has become established that lowering blood pressure in high-risk patients is a highly effective form of primary prevention for stroke. Over the subsequent 25 years, over 30 large clinical trials have extended these initial observations to allow us to conclude that treatment of mild, moderate or severe hypertension, and isolated systolic hypertension in the elderly, all produce important absolute benefits. In addition, excellent specific evidence of benefit is now accumulating for certain groups of normotensive patients, including those with previous stroke, and those with established cardiovascular disease. Although the importance of vigorous antihypertensive therapy for the primary and secondary prevention of stroke is increasingly clear, a large number of unanswered questions remain. For example, while it is apparent that diuretics, beta-blockers, calcium channel blockers (CCBs) and angiotensin converting enzyme (ACE) inhibitors are all effective antihypertensive agents, the question remains as to which drug, or combination of drugs, is best for which patients. The results of several ongoing comparative trials of different drug regimens, including the Anglo-Scandinavian Cardiac Outcomes Trial (ASCOT), may elucidate this further. PMID- 12740553 TI - Clinical trials: Evidence and unanswered questions--hyperlipidaemia. AB - It is now clear that the management of hypercholesterolaemia is important for the reduction of morbidity and mortality caused by cerebrovascular and coronary events. The landmark Scandinavian Simvastatin Survival Study was the first to show conclusively that lipid-lowering therapy with statins reduces the incidence of stroke. Subsequent trials, undertaken in a variety of different patient populations, have confirmed that statin therapy reduces the incidence of stroke by approximately one-third. This important benefit has been observed in men and women, the young and the elderly, and also in subjects with diabetes mellitus. In the recent Heart Protection Study, which recruited "high-risk" vascular subjects, stroke risk reduction was demonstrated even among those subjects considered to have "low" low-density lipoprotein (LDL) cholesterol levels. The benefits of statin therapy in stroke have been attributed to reductions in cholesterol and to other non-lipid-lowering effects of statins. Ongoing clinical trials such as TNT (Treating to New Targets) and IDEAL (Incremental Decrease in Endpoints through Aggressive Lipid lowering) will test the "lower is better" hypothesis. Using statins to lower LDL cholesterol to levels that are below current guidelines will provide additional benefits in stroke risk reduction. Most of the data on cholesterol reduction and cerebrovascular events have been derived from studies of patients with documented coronary heart disease (CHD). The ongoing SPARCL (Stroke Prevention by Aggressive Reduction in Cholesterol Levels) trial will examine the benefits of LDL cholesterol lowering in patients with previous stroke or transient ischaemic attack (TIA), but no history of coronary problems. PMID- 12740555 TI - [Useful knowledge for diagnostic imaging of oral region]. PMID- 12740554 TI - Blood pressure and lipid lowering in the prevention of stroke: a note to neurologists. AB - Stroke is the leading cause of adult disability and dependency in western society. Despite the determined efforts of basic science and clinical investigators, neuroprotective therapies for acute stroke have yet to be realised. Stroke prevention, therefore, remains the key route for reducing morbidity and mortality. Hypertension and hypercholesterolaemia are the most important modifiable risk factors for stroke. Several recent landmark studies have shown that lipid lowering with statins can reduce the risk of ischaemic stroke, as well as coronary heart disease. In addition, clinical trials evaluating the effects of blood pressure lowering have shown that antihypertensive agents such as angiotensin-converting enzyme (ACE) inhibitors, calcium channel blockers (CCBs) and angiotensin II receptor antagonists can reduce stroke risk. Accumulating evidence suggests certain antihypertensive agents such as CCBs might also prevent the formation and progression of carotid atheroma, independently of their blood-pressure-lowering effects. It follows that rigorous identification and targeting of high- risk or stroke-prone individuals for blood pressure and lipid-lowering interventions should be of practical importance to all physicians involved in the management of stroke. PMID- 12740556 TI - [Fourier transform for MRI]. PMID- 12740557 TI - [A questionnaire survey for ordering system in radiological examination]. PMID- 12740558 TI - [Research on the measurement of patient dose and protection in IVR]. PMID- 12740559 TI - [Activities of the 2002 Standardization Committee for radiological technology]. PMID- 12740560 TI - [Usefulness of low tube operating potential (TOP) for improved detection of minimally contrast-enhanced nodules CT]. AB - PURPOSE: We recently observed that hepatic nodules which were only minimally enhanced on dynamic CT were much better visualized using a low TOP in spite of some increase of the background noise. The purpose of this study is to evaluate the potential superiority of low TOP in detecting minimally contrast-enhanced hepatic nodules on CT. METHODS AND MATERIALS: A phantom was made of anger and contrast medium. Four anger columns were made four different concentrations of contrast media (0.8%, 1.0% 1.5% 1.8%). The phantom was scanned using different TOP (80, 100, 120, 140 kVp). Detectability of the columns were also evaluated contrast to noise ratio (CNR) and visually by radiologists in a doubleblind fashion. Weighted computed tomography dose index (CTDIw) was compared with different TOP. In the clinical study, seven patients were studied. The diagnostic usefulness of a TOP of 80 kVp was compared with that of 120 kVp in same patient by Dynamic CT which has better detectability of minimally contrast enhancing nodules. RESULT: With using lower TOP, CNR was increased. On visual evaluation by radiologists, images taken at 80 kVp achieved the highest detectability of the columns. CTDIw associated with the 80, 100, 120, 140 kVp at one scan were 5.78 mGy, 6.64 mGy, 8.13 mGy, 11.02 mGy, respectively. In the clinical study a lesion could be detected clearly with a low TOP of 80 kVp though the existence of the lesion could not be detected at 120 kVp. The existence of the HCC was proven with DSA, CTA and CTAP. CONCLUSION: It was demonstrated in the phantom study that if up-to-date equipment is used, CT operated at a TOP of 80 kVp has potentially better detectability of minimally contrast-enhanced nodules than 120 kVp. PMID- 12740561 TI - [Development of computerized method for automated classification of the body parts in digital radiographs]. AB - PURPOSE: In picture archiving and communication systems (PACS), the information on the body parts included in radiographs is often not or incorrectly recorded in an image header. In order to apply the computer-aided diagnosis (CAD) system in the PACS environment, the body parts in radiographs need to be recognized correctly by computer. The purpose of this study is to develop a computerized method for correctly classifying the body parts in digital radiographs based on a template matching technique. METHODS/MATERIALS: The image database used in this study was 1032 digital radiographs (14 x 17 inches) obtained with a computed radiography, and included 505 chest of postetroanterior view, 39 chest of lateral view, 241 abdomen, 108 pelvis, 10 upper limbs, 125 lower limbs, and 4 thoracic spine. In this method, test images were classified into four body parts, i.e., (1) chest, (2) abdomen, (3) pelvis, and (4) upper/lower limbs and thoracic spine. This computerized method was tested with 852 images, since 180 images were employed for creation of 98 templates, which represented the average radiographs for various body parts. Our approach was to examine the similarity of a given test image with templates by use of the cross-correlation values as the similarity measures. The body part of the test image was identified as the body part in the template yielding the maximum correlation value. Our method consisted of the following five steps. First, test images were classified into one of three groups; i.e. 1) chest and abdomen, 2) pelvis, and 3) upper/lower limbs and thoracic spine by using the templates obtained from images with the average size and position. Second, the remaining uncertain images were classified by using additional templates in various directions. Third, the chest and abdomen group was separated into two subgroups; i.e.chest and abdomen. Fourth, in order to classify some uncertain images, templates were shifted horizontally and vertically. Fifth, outer pixels of templates were eliminated to avoid the misclassification due to x-ray collimation. RESULTS: Our preliminary results indicated that the body parts for 850 cases (99.8%) were correctly classified with our method. CONCLUSIONS: This method would be useful for automated identification of the body parts in radiographs when various CAD systems would be implemented in the PACS environment. PMID- 12740562 TI - [PET technology applied for FDG based examination--in consideration of the approval by the health insurance system-Part 2]. PMID- 12740563 TI - [Study of elliptical centric view ordering technique with spectrally selected inversion recovery pulse (spec-IR pulse)]. AB - Among several techniques for contrast-enhanced MRA, the elliptical centric view ordering method, originally developed at the Mayo Clinic, is a promising one. It has been difficult to apply a fat-suppression prepulse, mainly because the conventional fat suppression method requires a longer acquisition time during sampling of the low-frequency domain in k-space, and it causes severe image distortion due to the great change in longitudinal magnetization derived from fat tissue. We developed a novel method to append fat saturation to the elliptical centric view ordering technique, and assessed the feasibility of its use. Our method is to apply fat-saturation pulses only at selected sampling points when any gradient is applied in the slice-encoding direction. In this way, we achieved efficient suppression of fat-derived signal within a relatively short time, comparable to that of the conventional fat-saturation method, and succeeded in minimizing artifacts. PMID- 12740564 TI - [Low contrast detectability of a new CR system with two-sided reading]. AB - A new imaging plate (IP) with a transparent support and reading system that can detect emissions from both sides of the IP has been developed and has already been introduced in some facilities. In this study, low contrast threshold detectability was investigated experimentally for a CR system with a two-sided reading system (new CR) and for a conventional CR system. Images of a Contrast Detail phantom were obtained with the new and conventional CR systems at dose levels corresponding to 26%, 49%, 82%, 103%, and 164% of the dose used for the screen-film system. Using an observer performance study of Contrast-Detail phantom images, the threshold contrast of disk-shaped objects ranging from 0.3 mm to 4.0 mm in diameter was determined. We also calculated image quality figure (IQF) from the results of observation. The new CR system showed significantly better contrast detectability than the conventional CR system. The use of a new CR system provided a reduction of approximately 25% in radiographic dose while providing comparable IQF. PMID- 12740565 TI - [Kinetic radiography and functional analysis of the temporomandibular joint (TMJ)]. AB - To develop a method of kinetic radiography and a computer-aided diagnosis (CAD) system for quantitative evaluation of the temporomandibular joint (TMJ), dynamic images of the TMJ from one healthy volunteer were obtained by fluoroscopy in the lateral view on the right and left sides. The accumulated image subtraction technique extracted the condyle in each image. A sequential similarity detection algorithm (SSDA) was employed to trace the movement path and the velocity of the condyle. The shape of the path of the right condyle was smoother than that of the left condyle. The size of the maximum vertical and horizontal movements of the condyle were 4.6+/-0.1 mm and 15.0+/-0.2 mm, respectively. The velocity of the movement of the condyle was higher in the area close to the articular eminence than in any other area during the opening and closing of the mouth. This CAD system will contribute to the kinetic analysis of the TMJ for screening, follow up study, and informed consent, providing speed, quantitation, and cost effectiveness. PMID- 12740566 TI - [Patient absorbed dose in coronary angiography determined by the flat panel digital detector X-ray system]. AB - The INNOVA 2000, an all-digital cardiovascular X-ray system with flat panel detector, is equipped with a monitoring function that makes it possible to track a patient's absorbed dose by displaying the real-time presumed absorbed dose. We verified this dose monitoring system and evaluated how it is affected by various parameters. We also compared the INNOVA 2000 to a conventional machine, the Advantx LC. The average absorbed dose of the INNOVA 2000 was 1,066 mGy, while that of the Advantx LC was calculated to be 2,028 mGy. Dose reduction with the INNOVA 2000 was 76% at Low mode and 52% even at Normal mode. The INNOVA provides an advantage in lowering absorbed dose, even considering that it has a rectangular image intensifier (I.I.) versus the Advantx LC's round I.I. This comparison was made by cine and digital angiography. PMID- 12740567 TI - Beta-amyloid secretases and beta-amloid degrading enzyme expression in lens. AB - PURPOSE: Beta- and gamma-secretases are proteases involved in the processing of the Alzheimer precursor protein (AbetaPP) that releases the transmembrane beta amyloid fragment (Abeta), associated with age-dependent disease in lens and brain. Gamma-secretase is a protein complex containing Presenilin and Nicastrin proteins, which also processes Notch and other receptors involved in the eye and lens development. Neprilysin (NEP), a major protease involved in degrading Abeta, acts with beta- and gamma-secretases to regulate steady-state levels of Abeta. Previously, we demonstrated AbetaPP and Presenilin expression and processing in the lens and demonstrated cell degeneration in classic Alzheimer disease (AD) transgenic and systemic oxidative stress animal models, suggesting that additional AbetaPP processing proteins are also present in the lens. Here we investigate lens expression of beta-secretases, nicastrin and NEP proteins, and compare their protein distribution to Notch and Presenilin in lens. METHODS: RT PCR was used to analyze mRNA transcripts. Immunoblots and immunohistochemistry were used to examine the protein expression and distribution of secretase and Abeta degrading proteins, as well as Presenilin and Notch proteins in mouse lenses. RESULTS: Beta-acting cleaving enzymes, BACE (BACE1) and BACE2, Nicastrin, Presenilins, Notch and NEP are expressed in the lens. In situ examination of protein distribution in lens indicates expression of each of these proteins is upregulated in peripheral elongating fiber cells at the lens equatorial margin and overlaps with Notch and Presenilin proteins, and also with the distribution of AbetaPP and Abeta proteins demonstrated in a previous study. Neprilysin exon 1 4 splicing, previously described as diagnostic for neuronal expression, also occurs in lens. CONCLUSIONS: BACE, BACE2, Nicastrin and NEP are expressed primarily in elongating peripheral fiber cells, overlapping with Notch, Presenilin, and AbetaPP protein distribution in lens, consistent with their role in regulating Notch and AbetaPP ectodomain shedding in lens. Lens expression of beta- and gamma-secretases together with NEP suggests these proteins may also regulate Abeta turnover in the lens. The presence of Abeta processing and degrading proteases in lens provides further evidence that Alzheimer-related cell biology is fundamentally involved in lens development, and provides additional evidence that mechanisms of Alzheimer pathophysiology can contribute to lens degeneration, suggesting further that therapeutics targeting Abeta proteases may be applicable to lens degenerative disease. PMID- 12740568 TI - Isolation and characterization of murine retinal endothelial cells. AB - PURPOSE: To isolate and characterize primary retinal endothelial cells (REC) from wild type and transgenic mice to facilitate the study of their properties in vitro. METHODS: REC were isolated from wild type or transgenic-immortomouse by collagenase digestion of retina and affinity purification using magnetic beads coated with platelet/endothelial cell adhesion molecule-1 (anti-PECAM-1). The bound cells were plated on fibronectin-coated wells and expanded. The REC were characterized for expression and localization of endothelial cell markers by fluorescence-activated cell sorting (FACS) analysis and indirect immunofluorescence staining. The ability of these cells to form capillary like networks was assessed on Matrigel while the migration properties were examined in wound closure assays. RESULTS: Isolation of REC from mouse has been very difficult and has not been previously reported. Here, we describe a method for isolation of retinal endothelial cells from wild type and thrombospondin-1 deficient (TSP1-/-) immortomice. Our results indicate that nearly 100% of selected cells express the endothelial cell marker PECAM-1 and vascular endothelial-cadherin (VE-cadherin). The cells were successfully passaged and maintained in culture for several months without a significant loss in expression of endothelial cell markers. The wild type REC, like most primary endothelial cells, formed capillary-like networks on Matrigel. The ability of the REC from TSP1-/- mice to form capillary-like networks on Matrigel was severely compromised. This may be attributed, at least in part, to the enhanced migratory and less differentiated phenotype of these cells. CONCLUSIONS: The retinal endothelial cells can be readily obtained from wild type and transgenic mice, which facilitate the comparison and identification of the physiologic role of specific genes in endothelial cell function. PMID- 12740569 TI - Pigment epithelium-derived factor regulates the vasculature and mass of the prostate and pancreas. AB - Angiogenesis sustains tumor growth and metastasis, and recent studies indicate that the vascular endothelium regulates tissue mass. In the prostate, androgens drive angiogenic inducers to stimulate growth, whereas androgen withdrawal leads to decreased vascular endothelial growth factor, vascular regression and epithelial cell apoptosis. Here, we identify the angiogenesis inhibitor pigment epithelium-derived factor (PEDF) as a key inhibitor of stromal vasculature and epithelial tissue growth in mouse prostate and pancreas. In PEDF-deficient mice, stromal vessels were increased and associated with epithelial cell hyperplasia. Androgens inhibited prostatic PEDF expression in cultured cells. In vivo, androgen ablation increased PEDF in normal rat prostates and in human cancer biopsies. Exogenous PEDF induced tumor epithelial apoptosis in vitro and limited in vivo tumor xenograft growth, triggering endothelial apoptosis. Thus, PEDF regulates normal pancreas and prostate mass. Its androgen sensitivity makes PEDF a likely contributor to the anticancer effects of androgen ablation. PMID- 12740570 TI - Targeting exogenous genes to tumor angiogenesis by transplantation of genetically modified hematopoietic stem cells. AB - Angiogenic tumor vessels are promising targets for the activity and the selective delivery of cancer therapeutics. The bone marrow contributes different cell types to the tumor stroma, including hematopoietic cells and, as recently suggested, vascular endothelial cells (ECs). Thus, transplantation of genetically modified bone marrow progenitors may represent a vehicle for the transport of gene therapy to tumors. We transduced bone marrow progenitors with lentiviral vectors expressing genes from transcription-regulatory elements of Tie2/Tek gene. When tumors were grown in the transplanted mice, the new vector marked a distinct hematopoietic population that 'homed' to the tumor and closely interacted with vascular ECs at the tumor periphery. These Tie2-expressing mononuclear (TEM) cells had a distinguishable phenotype and were present selectively at angiogenic sites. Unexpectedly, we did not find bone marrow-derived ECs in tumor vessels when we transplanted bone marrow progenitors constitutively expressing a marker gene from the Tie2 or ubiquitously active promoters. By delivering a 'suicide' gene, we selectively eliminated the TEM cells and achieved substantial inhibition of angiogenesis and slower tumor growth without systemic toxicity. Thus, TEM cells may account for the proangiogenic activity of bone marrow-derived cells in tumors, may represent a new target for drug development and may provide the means for selective gene delivery and targeted inhibition of tumor angiogenesis. PMID- 12740571 TI - Prostaglandin E2-EP4 signaling initiates skin immune responses by promoting migration and maturation of Langerhans cells. AB - Antigen-specific immune responses in the skin are initiated by antigen uptake into Langerhans cells and the subsequent migration of these cells to draining lymph nodes. Although prostaglandin E2 (PGE2) is produced substantially in skin exposed to antigen, its role remains unclear. Here we show that although Langerhans cells express all four PGE receptor subtypes, their migration to regional lymph nodes was decreased only in EP4-deficient (Ptger4-/-) mice and in wild-type mice treated with an EP4 antagonist. An EP4 agonist promoted the migration of Langerhans cells, increased their expression of costimulatory molecules and enhanced their ability to stimulate T cells in the mixed lymphocyte reaction in vitro. Contact hypersensitivity to antigen was impaired in Ptger4-/- mice and in wild-type mice treated with the EP4 antagonist during sensitization. PGE2-EP4 signaling thus facilitates initiation of skin immune responses by promoting the migration and maturation of Langerhans cells. PMID- 12740572 TI - Attenuation of levodopa-induced dyskinesia by normalizing dopamine D3 receptor function. AB - In monkeys rendered parkinsonian with 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP), expression of the dopamine D3 receptor was decreased. However, levodopa-induced dyskinesia (LID), similar to the debilitating and pharmacoresistant involuntary movements elicited after long-term treatment with levodopa in patients with Parkinson disease (PD), was associated with overexpression of this receptor. Administration of a D3 receptor-selective partial agonist strongly attenuated levodopa-induced dyskinesia, but left unaffected the therapeutic effect of levodopa. In contrast, attenuation of dyskinesia by D3 receptor antagonists was accompanied by the reappearance of PD like symptoms. These results indicated that the D3 receptor participated in both dyskinesia and the therapeutic action of levodopa, and that partial agonists may normalize D3 receptor function and correct side effects of levodopa therapy in patients with PD. PMID- 12740573 TI - Pneumococcal vaccination decreases atherosclerotic lesion formation: molecular mimicry between Streptococcus pneumoniae and oxidized LDL. AB - During the progression of atherosclerosis, autoantibodies are induced to epitopes of oxidized low-density lipoprotein (oxLDL) and active immunization of hypercholesterolemic mice with oxLDL ameliorates atherogenesis. We unexpectedly found that many autoantibodies to oxLDL derived from 'naive' atherosclerotic mice share complete genetic and structural identity with antibodies from the classic anti-phosphorylcholine B-cell clone, T15, which protect against common infectious pathogens, including pneumococci. To investigate whether in vivo exposure to pneumococci can affect atherogenesis, we immunized Ldlr(-/-) mice with Streptococcus pneumoniae. This induced high circulating levels of oxLDL-specific IgM and a persistent expansion of oxLDL-specific T15 IgM-secreting B cells primarily in the spleen, which were cross-reactive with pneumococcal determinants. Pneumococcal immunization decreased the extent of atherosclerosis, and plasma from these mice had an enhanced capacity to block the binding of oxLDL to macrophages. These studies show molecular mimicry between epitopes of oxLDL and S. pneumoniae and indicate that these immune responses can have beneficial effects. PMID- 12740574 TI - Resistance to CpG DNA-induced autoimmunity through tolerogenic B cell antigen receptor ERK signaling. AB - CpG sequences in self-DNA are an important potential trigger for autoantibody secretion in systemic lupus and other systemic autoimmune disorders. It is not known how this ubiquitous threat may be controlled by active mechanisms for maintaining self tolerance. Here we show that two distinct mechanisms oppose autoantibody secretion induced by CpG DNA in anergic B cells that are constantly binding self-antigen. Uncoupling of the antigen receptor (BCR) from a calcineurin dependent pathway prevents signals that synergize with CpG DNA for proliferation. The BCR does not become desensitized by activating the extracellular response kinase (ERK) MAP kinase pathway, however, and continuous self-antigen signaling to ERK inhibits CpG DNA-induced plasma cell differentiation. These two mechanisms seem to act as a general control against autoantibody production elicited by Toll like receptors, and their regulation of T cell-independent responses to Toll-like receptor 9 (TLR9) is probably crucial for resistance to systemic autoimmunity. PMID- 12740575 TI - NKG2D-DAP10 triggers human NK cell-mediated killing via a Syk-independent regulatory pathway. AB - The immune recognition receptor complex NKG2D-DAP10 on natural killer cells is stimulated by specific ligands carried on virus-infected and malignant cells. Because DAP10 does not have an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic tail, its ability to trigger killing has been debated. Here we show that a crucial Tyr-Ile-Asn-Met amino acid motif in the cytoplasmic tail of DAP10 couples receptor stimulation to the downstream activation of phosphatidylinositol 3-kinase, Vav1, Rho family GTPases and phospholipase C. Unlike that of ITAM-containing receptors, the activation of NKG2D-DAP10 proceeds independently of Syk family protein tyrosine kinases. Yet the signals initiated by NKG2D-DAP10 are fully capable of inducing killing. Our findings identify a previously unknown mechanism by which receptor complexes that lack ITAM motifs can trigger lymphocyte activation. PMID- 12740576 TI - NKG2D triggers cytotoxicity in mouse NK cells lacking DAP12 or Syk family kinases. AB - In activated mouse natural killer (NK) cells, the NKG2D receptor associates with two intracellular adaptors, DAP10 and DAP12, which trigger phosphatidyl inositol 3 kinase (PI3K) and Syk family protein tyrosine kinases, respectively. Here we show that cytotoxicity, but not cytokine production, is triggered by NKG2D in activated NK cells lacking either DAP12 or the Syk family members Syk and ZAP70. Inhibition of PI3K blocks this cytotoxicity, suggesting that the DAP10-PI3K pathway is sufficient to initiate NKG2D-mediated killing of target cells. Our results highlight signaling divergence in the effector functions of NKG2D and indicate that alternative associations between a receptor and its adaptors may provide a single receptor with a dual 'on-switch', giving mouse NK cells more choices through which to trigger cytotoxicity. PMID- 12740577 TI - The role of DNA methylation in setting up chromatin structure during development. AB - DNA methylation inhibits gene expression in animal cells, probably by affecting chromatin structure. Biochemical studies suggest that this process may be mediated by methyl-specific binding proteins that recruit enzymatic machinery capable of locally altering histone modification. To test whether DNA methylation actually has a role in the assembly of chromatin during normal development, we used cell transfection and a transgene construct genetically programmed to be either methylated or unmethylated in all cell types of the mouse. Chromatin immunoprecipitation (ChIP) analysis shows that the presence of DNA methylation brings about the deacetylation of histone H4 and methylation of Lys9 of histone H3 (H3 Lys9) and prevents methylation of Lys4 of histone H3 (H3 Lys4), thus generating a structure identical to that of methylated sequences in the genome. These results indicate that the methylation pattern established in early embryogenesis is profoundly important in setting up the structural profile of the genome. PMID- 12740578 TI - Trans allele methylation and paramutation-like effects in mice. AB - In mammals, imprinted genes have parent-of-origin-specific patterns of DNA methylation that cause allele-specific expression. At Rasgrf1 (encoding RAS protein-specific guanine nucleotide-releasing factor 1), a repeated DNA element is needed to establish methylation and expression of the active paternal allele. At Igf2r (encoding insulin-like growth factor 2 receptor), a sequence called region 2 is needed for methylation of the active maternal allele. Here we show that replacing the Rasgrf1 repeats on the paternal allele with region 2 allows both methylation and expression of the paternal copy of Rasgrf1, indicating that sequences that control methylation can function ectopically. Paternal transmission of the mutated allele also induced methylation and expression in trans of the normally unmethylated and silent wild-type maternal allele. Once activated, the wild-type maternal Rasgrf1 allele maintained its activated state in the next generation independently of the paternal allele. These results recapitulate in mice several features in common with paramutation described in plants. PMID- 12740579 TI - Module networks: identifying regulatory modules and their condition-specific regulators from gene expression data. AB - Much of a cell's activity is organized as a network of interacting modules: sets of genes coregulated to respond to different conditions. We present a probabilistic method for identifying regulatory modules from gene expression data. Our procedure identifies modules of coregulated genes, their regulators and the conditions under which regulation occurs, generating testable hypotheses in the form 'regulator X regulates module Y under conditions W'. We applied the method to a Saccharomyces cerevisiae expression data set, showing its ability to identify functionally coherent modules and their correct regulators. We present microarray experiments supporting three novel predictions, suggesting regulatory roles for previously uncharacterized proteins. PMID- 12740580 TI - Distinct spatial frequency sensitivities for processing faces and emotional expressions. AB - High and low spatial frequency information in visual images is processed by distinct neural channels. Using event-related functional magnetic resonance imaging (fMRI) in humans, we show dissociable roles of such visual channels for processing faces and emotional fearful expressions. Neural responses in fusiform cortex, and effects of repeating the same face identity upon fusiform activity, were greater with intact or high-spatial-frequency face stimuli than with low frequency faces, regardless of emotional expression. In contrast, amygdala responses to fearful expressions were greater for intact or low-frequency faces than for high-frequency faces. An activation of pulvinar and superior colliculus by fearful expressions occurred specifically with low-frequency faces, suggesting that these subcortical pathways may provide coarse fear-related inputs to the amygdala. PMID- 12740581 TI - Dopamine gates LTP induction in lateral amygdala by suppressing feedforward inhibition. AB - Fear conditioning involves the induction of long-term potentiation (LTP) of excitatory synaptic transmission in the lateral amygdala, a brain structure which is tightly controlled by GABAergic inhibition. Here we show that dopamine gates the induction of LTP in the mouse lateral amygdala by suppressing feedforward inhibition from local interneurons. Our findings provide a cellular mechanism for the dopaminergic modulation of fear conditioning and indicate that suppression of feedforward inhibition represents a key mechanism for the induction of associative synaptic plasticity in the lateral amygdala. PMID- 12740582 TI - Color brings relief to human vision. AB - In natural scenes, chromatic variations, and the luminance variations that are aligned with them, mainly arise from surfaces such as flowers or painted objects. Pure or near-pure luminance variations, on the other hand, mainly arise from inhomogeneous illumination such as shadows or shading. Here, I provide evidence that knowledge of these color-luminance relationships is built into the machinery of the human visual system. When a pure-luminance grating is added to a differently oriented chromatic grating, the resulting 'plaid' appears to spring into three-dimensional relief, an example of 'shape-from-shading'. By psychophysical measurements, I found that the perception of shape-from-shading in the plaid was triggered when the chromatic and luminance gratings were not aligned, and suppressed when the gratings were aligned. This finding establishes a new role for color vision in determining the three-dimensional structure of an image: one that exploits the natural relationships that exist between color and luminance in the visual world. PMID- 12740583 TI - Rapid global shifts in natural scenes block spiking in specific ganglion cell types. AB - The mammalian retina contains more than a dozen different ganglion cell types, each with dendrites ramifying at different strata within the inner plexiform layer (IPL) and each carrying a unique representation of the visual world. We studied the inhibitory and excitatory inputs, as well as the spiking output, of each of the rabbit retinal ganglion cell type during rapid global shifts in 'natural' videos designed to mimic saccadic eye movements. These shifts generated stratum-specific transient inhibitory activity, affecting only those ganglion cells whose dendrites ramify within the central strata of the IPL. The inhibition was GABA-mediated, acted both pre- and postsynaptically and was fast enough to either prevent or delay spiking. These findings indicate that the fast, transient elevation of visual threshold during rapid shifts in scene has a significant retinal component. PMID- 12740584 TI - High-throughput classification of yeast mutants for functional genomics using metabolic footprinting. AB - Many technologies have been developed to help explain the function of genes discovered by systematic genome sequencing. At present, transcriptome and proteome studies dominate large-scale functional analysis strategies. Yet the metabolome, because it is 'downstream', should show greater effects of genetic or physiological changes and thus should be much closer to the phenotype of the organism. We earlier presented a functional analysis strategy that used metabolic fingerprinting to reveal the phenotype of silent mutations of yeast genes. However, this is difficult to scale up for high-throughput screening. Here we present an alternative that has the required throughput (2 min per sample). This 'metabolic footprinting' approach recognizes the significance of 'overflow metabolism' in appropriate media. Measuring intracellular metabolites is time consuming and subject to technical difficulties caused by the rapid turnover of intracellular metabolites and the need to quench metabolism and separate metabolites from the extracellular space. We therefore focused instead on direct, noninvasive, mass spectrometric monitoring of extracellular metabolites in spent culture medium. Metabolic footprinting can distinguish between different physiological states of wild-type yeast and between yeast single-gene deletion mutants even from related areas of metabolism. By using appropriate clustering and machine learning techniques, the latter based on genetic programming, we show that metabolic footprinting is an effective method to classify 'unknown' mutants by genetic defect. PMID- 12740585 TI - Inhibition of hepatitis B virus in mice by RNA interference. AB - Hepatitis B virus (HBV) infection substantially increases the risk of chronic liver disease and hepatocellular carcinoma in humans. RNA interference (RNAi) of virus-specific genes has emerged as a potential antiviral mechanism. Here we show that RNAi can be applied to inhibit production of HBV replicative intermediates in cell culture and in immunocompetent and immunodeficient mice transfected with an HBV plasmid. Cotransfection with plasmids expressing short hairpin RNAs (shRNAs) homologous to HBV mRNAs induced an RNAi response. Northern and Southern analyses of mouse liver RNA and DNA showed substantially reduced levels of HBV RNAs and replicated HBV genomes upon RNAi treatment. Secreted HBV surface antigen (HBsAg) was reduced by 94.2% in cell culture and 84.5% in mouse serum, whereas immunohistochemical detection of HBV core antigen (HBcAg) revealed >99% reduction in stained hepatocytes upon RNAi treatment. Thus, RNAi effectively inhibited replication initiation in cultured cells and mammalian liver, showing that such an approach could be useful in the treatment of viral diseases. PMID- 12740586 TI - Global protein function prediction from protein-protein interaction networks. AB - Determining protein function is one of the most challenging problems of the post genomic era. The availability of entire genome sequences and of high-throughput capabilities to determine gene coexpression patterns has shifted the research focus from the study of single proteins or small complexes to that of the entire proteome. In this context, the search for reliable methods for assigning protein function is of primary importance. There are various approaches available for deducing the function of proteins of unknown function using information derived from sequence similarity or clustering patterns of co-regulated genes, phylogenetic profiles, protein-protein interactions (refs. 5-8 and Samanta, M.P. and Liang, S., unpublished data), and protein complexes. Here we propose the assignment of proteins to functional classes on the basis of their network of physical interactions as determined by minimizing the number of protein interactions among different functional categories. Function assignment is proteome-wide and is determined by the global connectivity pattern of the protein network. The approach results in multiple functional assignments, a consequence of the existence of multiple equivalent solutions. We apply the method to analyze the yeast Saccharomyces cerevisiae protein-protein interaction network. The robustness of the approach is tested in a system containing a high percentage of unclassified proteins and also in cases of deletion and insertion of specific protein interactions. PMID- 12740591 TI - Targeting Ras/ERK signaling in the striatum: will it help? PMID- 12740590 TI - Regulation of food intake: an 'old' actor plays a 'new' role. PMID- 12740587 TI - Discovering potent and selective reversible inhibitors of enzymes in complex proteomes. AB - To realize the promise of genomics-based therapeutics, new methods are needed to accelerate the discovery of small molecules that selectively modulate protein activity. Toward this end, advances in combinatorial synthesis have provided unprecedented access to large compound libraries of considerable structural complexity and diversity, shifting the bottleneck in drug discovery to the development of efficient screens for protein targets. Screening for reversible enzyme inhibitors typically requires extensive target-specific work, including protein expression and purification, as well as the development of specific substrate assays. Here we report a proteomic method for the discovery of reversible enzyme inhibitors that avoids these steps. We show that competitive profiling of a library of candidate serine hydrolase inhibitors in complex proteomes with activity-based chemical probes identifies nanomolar reversible inhibitors of several enzymes simultaneously, including the endocannabinoid degrading enzyme fatty acid amide hydrolase (FAAH), triacylglycerol hydrolase (TGH) and an uncharacterized membrane-associated hydrolase that lacks known substrates. The strategy tests inhibitors against numerous enzymes in parallel, assigning both potency and selectivity factors to each agent. In this way, promiscuous inhibitors were readily rejected in favor of equally potent compounds with 500-fold or greater selectivity for their targets. PMID- 12740592 TI - Dopamine and Parkinson's disease: is the killer in the house? PMID- 12740593 TI - Further evidence for a modulation of Novelty Seeking by DRD4 exon III, 5-HTTLPR, and COMT val/met variants. PMID- 12740594 TI - Glutamatergic mechanisms in addiction. AB - Traditionally, addiction research in neuroscience has focused on mechanisms involving dopamine and endogenous opioids. More recently, it has been realized that glutamate also plays a central role in processes underlying the development and maintenance of addiction. These processes include reinforcement, sensitization, habit learning and reinforcement learning, context conditioning, craving and relapse. In the past few years, some major advances have been made in the understanding of how glutamate acts and interacts with other transmitters (in particular, dopamine) in the context of processes underlying addiction. It appears that while many actions of glutamate derive their importance from a stimulatory interaction with the dopaminergic system, there are some glutamatergic mechanisms that contribute to addiction independent of dopaminergic systems. Among those, context-specific aspects of behavioral determinants (ie control over behavior by conditioned stimuli) appear to depend heavily on glutamatergic transmission. A better understanding of the underlying mechanisms might open new avenues to the treatment of addiction, in particular regarding relapse prevention. PMID- 12740595 TI - Psychotic symptoms in Alzheimer disease: evidence for a distinct phenotype. AB - Though efforts to identify the genetic etiology of Alzheimer disease (AD) have made substantial progress, to date only some of the genes contributing to AD risk have been identified. Utilization of more etiologically homogeneous subphenotypes represents one strategy to facilitate the identification of novel risk genes in complex disorders. In this review, we evaluate the hypothesis that psychotic symptoms, such as delusions and hallucinations, define a suitable subphenotype in AD patients for gene-mapping efforts. Psychotic symptoms occur in 40-60% of patients with AD and are associated with more severe cognitive deficits and a more rapidly deteriorating course. The presence of psychotic symptoms in AD confers increased risk of similar symptoms to affected siblings. Candidate gene association analyses and initial linkage analysis have yielded significant results. We discuss possible genetic models of psychotic symptoms in AD, and suggest strategies for further investigation. Identification of such genetic factors may facilitate gene-mapping studies for both AD and idiopathic psychoses. PMID- 12740596 TI - The dopamine transporter gene is associated with attention deficit hyperactivity disorder in a Taiwanese sample. AB - Genetic variation of the dopamine transporter gene (DAT1) is of particular interest in the study of attention-deficit hyperactivity disorder (ADHD), since stimulant drugs interact directly with the transporter protein. Association between ADHD and the 10-repeat allele of a 40-bp VNTR polymorphism that lies within the 3'-UTR of DAT1 was first reported in 1995, a finding that has been replicated in at least six independent samples from Caucasian populations. We analysed the DAT1 polymorphism in a sample of 110 Taiwanese probands with a DSM IV diagnosis of ADHD and found evidence of increased transmission of the 10 repeat allele using TRANSMIT (chi(2)=10.8, 1 d.f., p=0.001, OR=2.9, 95% CI 1.4 6.3). These data give rise to a similar odds ratio to that observed in Caucasian poplulations despite a far higher frequency of the risk allele in this Taiwanese population; 82.3% in the un-transmitted parental alleles and 94.5% in the ADHD probands. These data support the role of DAT1 in ADHD susceptibility among Asian populations. PMID- 12740597 TI - Candidate genes for anorexia nervosa in the 1p33-36 linkage region: serotonin 1D and delta opioid receptor loci exhibit significant association to anorexia nervosa. AB - Serotonergic and opioidergic neurotransmitter system alterations have been observed in people with eating disorders; the genes for the serotonin 1D receptor (HTR1D) and the opioid delta receptor (OPRD1) are found on chr1p36.3-34.3, a region identified by our group in a linkage analysis of anorexia nervosa (AN). These candidate genes were evaluated for sequence variation and for linkage and association of this sequence variation to AN in family and case : control data sets. Resequencing of the HTR1D locus and a portion of the OPRD1 locus identified novel SNPs and confirmed existing SNPs. Genotype assay development and genotyping of nine SNPs (four at HTR1D and five at OPRD1) was performed on 191 unrelated individuals fulfilling DSM-IV criteria (w/o amenorrhea criterion) for AN, 442 relatives of AN probands and 98 psychiatrically screened controls. Linkage analysis of these candidate gene SNPs with 33 microsatellite markers in families including relative pairs concordantly affected with restricting AN (N=37) substantially increased the evidence for linkage of this region to restricting AN to an NPL score of 3.91. Statistically significant genotypic, allelic, and haplotypic association to AN in the case : control design was observed at HTR1D and OPRD1 with effect sizes for individual SNPs of 2.63 (95% CI=1.21-5.75) for HTR1D and 1.61 (95% CI=1.11-2.44) for OPRD1. Using genotype data on parents and AN probands, three SNPs at HTR1D were found to exhibit significant transmission disequilibrium (P&<0.05). The combined statistical genetic evidence suggests that HTR1D and OPRD1 or linked genes may be involved in the etiology of AN. PMID- 12740598 TI - Tyrosine nitration of a synaptic protein synaptophysin contributes to amyloid beta-peptide-induced cholinergic dysfunction. AB - Amyloid beta (Abeta) is a critical factor involved in the pathogenesis of Alzheimer's disease (AD). We have previously demonstrated that continuous intracerebroventricular infusion of Abeta1-40 induced a time-dependent expression of the inducible nitric oxide (NO) synthase (iNOS) and an overproduction of NO in the rat hippocampus. The pathophysiological significance of the overproduction of NO on brain function was manifested by an impairment of nicotine-evoked acetylcholine(ACh) release and memory deficits.(4) Molecular mechanisms by which NO participates in the Abeta-induced brain dysfunction, however, remain to be determined. Here we show that chronic Abeta1-40 infusion caused a robust peroxynitrite formation and subsequent tyrosine nitration of proteins in the hippocampus. Immunoprecipitation and Western blot analyses further revealed that synaptophysin, a synaptic protein, was a main target of tyrosine nitration. Chronic infusion of Abeta1-40 resulted in an impairment of nicotine-evoked ACh release as analyzed by microdialysis. Daily treatment with the iNOS inhibitor aminoguanidine (AG) or the peroxynitrite scavenger uric acid (UA) prevented the tyrosine nitration of synaptophysin as well as the impairment of nicotine-evoked ACh release induced by Abeta. Our findings suggest that the tyrosine nitration of synaptophysin is related to Abeta-induced impairment of ACh release. PMID- 12740599 TI - Islet-brain1/C-Jun N-terminal kinase interacting protein-1 (IB1/JIP-1) promoter variant is associated with Alzheimer's disease. AB - Islet-brain1 (IB1) or c-Jun NH2 terminal kinase interacting protein-1 (JIP-1), the product of the MAPK8IP1 gene, functions as a neuronal scaffold protein to allow signalling specificity. IB1/JIP-1 interacts with many cellular components including the reelin receptor ApoER2, the low-density lipoprotein receptor related protein (LRP), kinesin and the Alzheimer's amyloid precursor protein. Coexpression of IB1/JIP-1 with other components of the c-Jun NH2 terminal-kinase (JNK) pathway activates the JNK activity; conversely, selective disruption of IB1/JIP-1 in mice reduces the stress-induced apoptosis of neuronal cells. We therefore hypothesized that IB1/JIP-1 is a risk factor for Alzheimer's disease (AD). By immunocytochemistry, we first colocalized the presence of IB1/JIP-1 with JNK and phosphorylated tau in neurofibrillary tangles. We next identified a 499A>G polymorphism in the 5' regulatory region of the MAPK8IP1 gene. In two separate French populations the -499A>G polymorphism of MAPK8IP1 was not associated with an increased risk to AD. However, when stratified on the +766C>T polymorphism of exon 3 of the LRP gene, the IB1/JIP-1 polymorphism was strongly associated with AD in subjects bearing the CC genotype in the LRP gene. The functional consequences of the -499A>G polymorphism of MAPK8IP1 was investigated in vitro. In neuronal cells, the G allele increased transcriptional activity and was associated with an enhanced binding activity. Taken together, these data indicate that the increased transcriptional activity in the presence of the G allele of MAPK8IP1 is a risk factor to the onset of in patients bearing the CC genotype of the LRP gene. PMID- 12740600 TI - Depressive-like behavior and stress reactivity are independent traits in a Wistar Kyoto x Fisher 344 cross. AB - Depression is a heritable disorder that is often precipitated by stress. Abnormalities of the stress-reactive hypothalamic-pituitary-adrenal (HPA) axis are also common in depressed patients. In animal models, the forced swim test (FST) is the most frequently used test of depressive-like behavior. We have used a proposed animal model of depression, the Wistar Kyoto (WKY) rat, to investigate the relationship as well as the mode of inheritance of FST behaviors and HPA measures. Through reciprocal breeding of WKY and F344 parent strains and brother sister breeding of the F1 generation, we obtained 486 F2 animals. Parent, F1 and F2 animals were tested in the FST. Blood samples were collected for determination of basal and stress (10-min restraint) plasma corticosterone (CORT) levels, and adrenal weights were measured. We found that all measures were heritable to some extent and that this heritability was highly sex dependent. Both correlation and factor analyses of the F2 generation data demonstrate that FST behavior and HPA axis measures are not directly related. Thus, the underlying genetic components of depressive-like behavior and HPA axis abnormalities are likely to be disparate in the segregating F2 generation of a WKY x F344 cross. PMID- 12740601 TI - A developmentally regulated and psychostimulant-inducible novel rat gene mrt1 encoding PDZ-PX proteins isolated in the neocortex. AB - Single or repeated exposure to psychostimulants such as amphetamines and cocaine after postnatal week 3 leads to an enduring enhancement in the psychotomimetic responses elicited by a subsequent challenge of a stimulant in rodents. This behavioral sensitization phenomenon has been considered to be the neural consequences of stimulant-induced alterations in gene expression in the brain after a critical period of postnatal development. Using a differential cloning technique, RNA arbitrarily primed PCR, we have now identified from the rat neocortex a novel and developmentally regulated methamphetamine (MAP)-inducible gene mrt1 (MAP responsive transcript 1). mrt1 encodes two major types of PDZ- and PX-domains containing proteins of approximately 62 kDa in size with different carboxy termini, Mrt1a and Mrt1b. The mrt1 mRNAs for Mrt1a, mrt1a, and for Mrt1b, mrt1b, are predominantly expressed in various brain regions and the testes, respectively. Acute MAP injection upregulated mrt1b expression in the neocortex after postnatal week 3 in a D1 receptor antagonist-sensitive manner without affecting mrt1a expression. This upregulation was mimicked by another stimulant, cocaine, whereas pentobarbital and D1 antagonist failed to change the mrt1b transcript levels. Moreover, repeated daily treatment of MAP, but not MAP plus D1 antagonist, for 5 days caused an augmentation of the basal expression of mrt1b 2 and 3 weeks after the drug discontinuation. These late-developing, cocaine crossreactive, D1 antagonist-sensitive and long-term regulations of mrt1b by MAP are similar to the pharmacological profiles of stimulant-induced behavioral sensitization, and therefore may be associated with the initiation and/or maintenance of the long-term neuronal adaptation. PMID- 12740602 TI - Linkage of schizophrenia with chromosome 1q loci in Taiwanese families. AB - A positive linkage of schizophrenia with chromosome 1q loci has been reported in Caucasian patients. This study was designed to evaluate the linkage of schizophrenia with markers of the 1q22-44 region in 52 Taiwanese families with at least two affected siblings. In the region 1q22-31 (17.8 cM), marker D1S1679 had a maximal proportion (0.57, P=0.03) of shared identity by descent (IBD) under a narrow phenotype (DSM-IV schizophrenia only). In the region 1q42-44 (26.8 cM), the marker D1S251, located near the breakpoint of a balanced translocation t (1;11) (q42.1;q14.3) segregated with schizophrenia, and also near the neurodevelopment-related 'Disrupted in Schizophrenia 1' gene, had a maximum NPL score of 1.73 (P=0.03) under the narrow phenotype model and 2.18 (P=0.01) under the broad phenotype model comprised of schizophrenia, schizoaffective disorder, and other nonaffective psychotic disorders as defined by DSM-IV criteria. The marker D1S2836 also had a maximal proportion (0.57, P=0.05) of shared IBD under the broad model. These findings may provide guidance for positional cloning studies on candidate genes in the 1q22-31 and 1q41-44 regions. PMID- 12740603 TI - Extrastriatal dopamine D 2/3 receptor density and distribution in drug-naive schizophrenic patients. AB - Several lines of studies have suggested the importance of cortical dopamine (DA) transmission in the pathophysiology of schizophrenia. The putative alteration of striatal D(2) receptor density in schizophrenia has been studied intensely, although extrastriatal DA activity may be more relevant for behavioral symptoms. The aim of this study was to explore extrastriatal D(2/3) density in drug-naive schizophrenic patients. We studied the extrastriatal D(2/3) receptor binding with a novel high-affinity single-photon emission tomography ligand epidepride in seven drug-naive schizophrenic patients and seven matched controls. The symptoms were rated with Positive and Negative Syndrome Scale for Schizophrenia. The findings indicated an extremely low D(2/3) receptor binding among patients in temporal cortex in both hemispheres when compared with controls (effect size 2.0 2.3), and the D(2/3) levels had negative correlations with general psychopathological (r from -0.86 to -0.90) and negative (r from -0.37 to -0.55) schizophrenic symptoms. These results support the previous hypothesis on dysfunction of mesocortical DA function behind the cognitive and negative symptoms in schizophrenia. PMID- 12740604 TI - Inhibition of intracellular hepatitis C virus replication by synthetic and vector derived small interfering RNAs. AB - Small interfering RNAs (siRNAs) efficiently inhibit gene expression by RNA interference. Here, we report efficient inhibition, by both synthetic and vector derived siRNAs, of hepatitis C virus (HCV) replication, as well as viral protein synthesis, using an HCV replicon system. The siRNAs were designed to target the 5' untranslated region (5' UTR) of the HCV genome, which has an internal ribosomal entry site for the translation of the entire viral polyprotein. Moreover, the 5' UTR is the most conserved region in the HCV genome, making it an ideal target for siRNAs. Importantly, we have identified an effective site in the 5' UTR at which approximately 80% suppression of HCV replication was achieved with concentrations of siRNA as low as 2.5 nM. Furthermore, DNA-based vectors expressing siRNA against HCV were also effective, which might allow the efficient delivery of RNAi into hepatocytes in vivo using viral vectors. Our results support the feasibility of using siRNA-based gene therapy to inhibit HCV replication, which may prove to be valuable in the treatment of hepatitis C. PMID- 12740605 TI - Identification of the fertility restoration locus, Rfo, in radish, as a member of the pentatricopeptide-repeat protein family. AB - Ogura cytoplasmic male sterility (CMS) in radish (Raphanus sativus) is caused by an aberrant mitochondrial gene, Orf138, that prevents the production of functional pollen without affecting female fertility. Rfo, a nuclear gene that restores male fertility, alters the expression of Orf138 at the post transcriptional level. The Ogura CMS/Rfo two-component system is a useful model for investigating nuclear-cytoplasmic interactions, as well as the physiological basis of fertility restoration. Using a combination of positional cloning and microsynteny analysis of Arabidopsis thaliana and radish, we genetically and physically delimited the Rfo locus to a 15-kb DNA segment. Analysis of this segment shows that Rfo is a member of the pentatricopeptide repeat (PPR) family. In Arabidopsis, this family contains more than 450 members of unknown function, although most of them are predicted to be targeted to mitochondria and chloroplasts and are thought to have roles in organellar gene expression. PMID- 12740606 TI - Regulation of neuronal SNARE assembly by the membrane. AB - In the neuron, SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) assembly acts centrally in driving membrane fusion, a required process for neurotransmitter release. In the cytoplasm, vesicular SNARE VAMP-2 (vesicle-associated membrane protein-2) engages with two plasma membrane SNAREs, syntaxin 1A and SNAP-25 (synaptosome-associated protein of 25 kDa), to form the core complex that bridges two membranes. Although various factors regulate SNARE assembly, the membrane also aids in regulation by trapping VAMP-2 in the membrane. Fluorescence and EPR analyses revealed that the insertion of seven C terminal core-forming residues into the membrane controls complex formation of the entire core region, even though the preceding 54 core-forming residues are fully exposed and freely moving. When two interfacial tryptophan residues in this region were replaced with hydrophilic serine residues, the mutation supported rapid complex formation. The results suggest that the membrane-proximal region of VAMP-2 is a regulatory module for SNARE assembly, providing new insights into calcium-triggered membrane fusion. PMID- 12740608 TI - Redox-sensitive extracellular gates formed by auxiliary beta subunits of calcium activated potassium channels. AB - An important step to understanding ion channels is identifying the structural components that act as the gates to ion movement. Here we describe a new channel gating mechanism, produced by the beta3 auxiliary subunits of Ca2+-activated, large-conductance BK-type K+ channels when expressed with their pore-forming alpha subunits. BK beta subunits have a cysteine-rich extracellular segment connecting two transmembrane segments, with small cytosolic N and C termini. The extracellular segments of the beta3 subunits form gates to block ion permeation, providing a mechanism by which current can be rapidly diminished upon cellular repolarization. Furthermore, this gating mechanism is abolished by reduction of extracellular disulfide linkages, suggesting that endogenous mechanisms may regulate this gating behavior. The results indicate that auxiliary beta subunits of BK channels reside sufficiently close to the ion permeation pathway defined by the alpha subunits to influence or block access of small molecules to the permeation pathway. PMID- 12740607 TI - X-ray snapshots of the maturation of an antibody response to a protein antigen. AB - The process whereby the immune system generates antibodies of higher affinities during a response to antigen (affinity maturation) is a prototypical example of molecular evolution. Earlier studies have been confined to antibodies specific for small molecules (haptens) rather than for proteins. We compare the structures of four antibodies bound to the same site on hen egg white lysozyme (HEL) at different stages of affinity maturation. These X-ray snapshots reveal that binding is enhanced, not through the formation of additional hydrogen bonds or van der Waals contacts or by an increase in total buried surface, but by burial of increasing amounts of apolar surface at the expense of polar surface, accompanied by improved shape complementarity. The increase in hydrophobic interactions results from highly correlated rearrangements in antibody residues at the interface periphery, adjacent to the central energetic hot spot. This first visualization of the maturation of antibodies to protein provides insights into the evolution of high affinity in other protein-protein interfaces. PMID- 12740609 TI - Supporting smoking cessation in pregnancy--action is urgently needed. PMID- 12740610 TI - New Zealand's Primary Health Care Strategy: what are the costs and how likely are the benefits? PMID- 12740611 TI - Improving the effectiveness of smoking cessation in primary care: lessons learned. AB - AIMS: The 'Smokescreen' smoking cessation programme was introduced in Christchurch in 1995, with an initial study showing six-month, self-reported quit rates of 10% and 17% (with a validated deception rate) in primary and secondary care settings. Substantial modifications were made to try to improve this rate in the primary care setting and the programme has been implemented widely. Our primary aim was to estimate programme utilisation and six-month quit rates for enrolled patients in this general primary care setting. We also aimed to use a wide range of patient, practice and environmental variables to estimate any predictive effect on outcome. METHODS: Prospective longitudinal cohort study. The nicotine replacement therapy (NRT) -based programme was implemented by Pegasus Health, an independent practitioner association (IPA) situated in the Christchurch urban area, to which the majority of Christchurch-based GPs belong. A cohort of 516 patients enrolling in the programme over a two-month period were contacted six months after their nominated quit date. The main outcome measure was the six-month, self-reported quit rate. RESULTS: Of the 516 participants, 334 (65%) were contacted by mail or telephone. The overall six-month quit rate was 36% (95% Confidence Interval (CI) 31-41). Univariate analysis initially showed duration of NRT (p = 0.03) and age band (p = 0.004) were significant predictors of quitting, while living with a smoker (p = 0.02), having made no previous quit attempts (p = 0.02) and having heart disease (p = 0.01) were all significant predictors of continued smoking at six months. Factors that did not predict whether respondents were smoking at six months included previous use of NRT, sex, ethnicity, who delivered the intervention, years of smoking, cigarette dose, and NZDep96 score. However, there was interaction between these factors as after multivariate analysis the only significant predictors of outcome were having others living in the house who smoked (odds ratio (OR) 0.55, 95% CI 0.33--0.93, p = 0.03) and having made no previous quit attempts (OR 0.29, 95% CI 0.12-0.71, p = 0.02). Both these factors were significantly associated with continuing to smoke. CONCLUSIONS: This programme compares favourably with six-month quit rates for NRT based programmes reported in the international literature of 14-22%. The effectiveness of an NRT-based smoking cessation programme in a general primary care setting appears to have been significantly enhanced by local adaptation, the flexibility of a primary-care-team approach and subsidisation of NRT, together with facilitation responsive to individual practice needs. The success of this programme in helping individual patients quit, as well as its successful implementation in a wide primary care setting, suggests General Practice can play an important role in smoking cessation in a country with a high burden of disease from smoking-related illnesses. The programme is congruent with the current, national, smoking cessation guidelines endorsed by the RNZCGP. Widespread adoption of this kind of model in IPA/primary health organisation (PHO) settings throughout New Zealand should be encouraged and supported. PMID- 12740612 TI - Factors that influence changes in smoking behaviour during pregnancy. AB - AIMS: This study explored characteristics of women who continue to smoke beyond the first trimester of pregnancy. METHODS: A cohort of 1283 pregnant women were surveyed at the time they registered with a maternity care provider, using a postal questionnaire. Women who reported they were ex-smokers were asked when they had stopped smoking. Data were analysed using logistic regression to identify socio-demographic variables associated with smoking and with stopping smoking. RESULTS: 829 (69.2%) women responded to the questionnaire. 183 (22.2%) reported smoking when they became pregnant. Forty nine (26.8%) of the women smoking at conception reported giving up in the first trimester. Factors significantly associated with stopping smoking in the first trimester were current employment (OR 2.37, 95% CI 1.16-4.85), first pregnancy (OR 2.18, 95% CI 1.11-4.28), and experiencing nausea during the pregnancy (OR 2.59, 95% CI 1.11 6.04). Women who held a community services card (OR 0.41, 95% CI 0.19-0.86), Maori women (OR 0.38, 95% CI 0.15-0.98) and women whose partners smoked (OR 0.35, 95% CI 0.17-0.70) were significantly less likely to have stopped smoking. CONCLUSION: Socioeconomically deprived women were more likely to continue to smoke beyond the first trimester of pregnancy and this needs to be taken into account in the provision of smoking cessation support. PMID- 12740613 TI - The cost to the New Zealand Government of providing 'free' primary medical care: an estimate based upon the Rand Health Insurance Experiment. AB - AIM: To estimate the likely cost to the New Zealand Government of providing universally free primary medical care. METHODS: Estimates of current government expenditure on various categories of health were obtained from Ministry of Health sources. Information from the Rand Health Insurance Experiment was used to estimate the changes in demand for general practitioner (GP) visits, pharmaceutical, laboratory and other referred services, and hospital services. The effects of a supplier response and complement (pharmaceutical) prices were also considered. RESULTS: Assuming that GPs act to control their patients' increasing demand for services, providing free primary care to all New Zealanders is likely to cost the Government between 435 million dollars and 592 million dollars (based upon 1998/99 year data). CONCLUSIONS: The difficulties and likely inexactness in making estimates of this sort are acknowledged; however, when considering such important changes in health policy, it is important to attempt to define likely costs (and benefits). Consideration of costs must go beyond simply estimating current private expenditure on primary medical care. PMID- 12740614 TI - Acute extensor hallucis longus tenosynovitis caused by gonococcal infection. PMID- 12740615 TI - An unusual cause of gastrointestinal obstruction: bezoar. PMID- 12740616 TI - Experiences with recreational drugs amongst first-year students at Auckland University. PMID- 12740617 TI - Severe acute respiratory syndrome: a storm in a teacup? PMID- 12740618 TI - Opportunities to improve cancer care in Australia and New Zealand. PMID- 12740619 TI - Experiences and preferences of general practitioners regarding continuing medical education: a qualitative study. AB - AIM: To explore the experiences and preferences of general practitioners (GPs) regarding their continuing medical education (CME). METHODS: Qualitative study using semi-structured interviews of twenty four GPs from Auckland and North Island rural areas assessing GPs' experiences and preferences. RESULTS: The need for CME was emphasised. Primary themes identified were: the value of personal interaction; the perception that CME that did not involve personal interaction was adjunctive; an opportunistic rather than needs-based approach to learning; a preference for succinct, evidenced-based, GP-focused content; and lack of time as a major barrier to obtaining optimal CME. CONCLUSIONS: Interactive formats are generally preferred, but identification of which elements of interactive formats facilitate learning is not established. Most GPs do not direct their CME according to the adult learning model. The challenge for CME providers is to provide avenues to facilitate needs identification and self-directed learning. PMID- 12740620 TI - Does the admitting officer system reduce the time taken to arrange an emergency admission to hospital? AB - AIM: Communication is an important feature of admitting acutely ill patients to hospital in both New Zealand and Australia. The mechanisms used to facilitate communication between the general practitioner (GP) and the admitting hospital differ between the two countries. The relative effectiveness of each of these systems has never been formally studied. Our aim was to compare the efficiency of the admitting officer system for arranging hospital admissions (used in Australia) with that of direct referral by the GP to the specialist registrar (the system used in New Zealand). METHODS: Five metropolitan hospitals of comparable size from each country were selected and the time taken to contact the relevant doctor in order to arrange admission of an acutely ill patient was documented. RESULTS: A total of 120 contact attempts were made, 60 in each country. The total time taken to contact the admitting doctor in order to arrange an emergency assessment for a patient was significantly longer in New Zealand than it was in Australia (p <0.05). CONCLUSIONS: When arranging an emergency patient assessment for a patient in the community, the total time taken to contact the appropriate doctor is less in Australia, where the admitting officer system is used, than it is in New Zealand, using the specialist registrar system. Consideration should be given by acute hospitals in New Zealand to streamlining their communications processes in order to minimise delays to referring doctors and their patients. PMID- 12740621 TI - Availability of urgent ultrasonography to emergency departments in New Zealand. AB - AIM: To determine the availability and use of urgent ultrasonography to emergency departments in New Zealand. METHODS: A questionnaire used in Canada was modified for New Zealand and mailed to all emergency department clinical directors in New Zealand who are listed by the Australasian Society for Emergency Medicine. RESULTS: The response rate was 92% (24/26). Most clinical directors found it difficult to obtain an ultrasound scan during 'off-hours'. Only one emergency department had the facility to get an ultrasound scan within 15 minutes. Seventy nine per cent of clinical directors felt that emergency physicians should perform ultrasonography, yet only 29% were actually doing so. Of those not performing ultrasonography, 47% had plans to do so in the future. CONCLUSIONS: Most clinical directors feel that it is difficult to get an urgent ultrasound scan and that emergency physicians should be performing ultrasonography. With the proper training and support most clinical directors had plans to use ultrasonography in the future. PMID- 12740622 TI - Major abdominal surgery in octogenarians. AB - AIMS: To evaluate long-term survival after major abdominal surgery in patients who are 80 years and over and to assess possible predictors of outcome: age, acute vs elective surgery, associated comorbidities and type of surgical procedure. METHODS: Patients who had surgery between 1 July 1997 and 1 July 1999 were reviewed. We reviewed 30-day mortality, major complications, hospital stay, intensive care unit (ICU) stay and long-term survival. General practitioners (GPs), family members or patients were contacted. RESULTS: One hundred and eighty patients had surgery, median age 84 (80-97), 115 females. Seventeen patients were lost to follow up. One hundred had an emergency procedure and 80 had elective procedures. Thirty-day mortality with emergency procedures was 29% and with elective operations, 7.5 % (p <0.0001). Overall morbidity was 33.3%. Long-term survival data were analysed using Kaplan-Meier survival curves and compared with the age-matched population obtained from Statistics New Zealand. They showed that long-term survival is similar between emergency and elective procedures after adjustment for peri-operative mortality. There was no survival difference between procedures. CONCLUSIONS: Elective surgery is generally well tolerated by the elderly. There is high in-hospital morbidity and mortality in the emergency group; however, long-term survival in those patients who leave hospital is not significantly different to the age-adjusted population. Age should not be used as the only criterion when deciding suitability for surgery in this age group. PMID- 12740623 TI - Intussusception of a normal appendix: how to avoid a right hemicolectomy. PMID- 12740624 TI - Septic arthritis due to a toxigenic strain of Corynebacterium diphtheriae gravis. PMID- 12740625 TI - The death of pharmacy as we know it. PMID- 12740626 TI - Last chance to fix the holes in proposed legislation for smoke-free environments. PMID- 12740627 TI - Psychiatric illness in primary care: whom should we treat? PMID- 12740628 TI - The health of New Zealand youth. PMID- 12740629 TI - The nature and prevalence of psychological problems in New Zealand primary healthcare: a report on Mental Health and General Practice Investigation (MaGPIe). AB - AIMS: This paper describes the methods used in a study of the prevalence and types of common mental disorders among patients attending New Zealand general practices, and reports some key findings from the first phase of the study. The study also aimed to determine the degree of associated disability and other factors influencing recognition, management, course and outcome of these disorders, and subsequent papers will address these issues. METHODS: General practitioners (GPs) were selected randomly. In the first phase of the study, all adult attenders at each practice on selected days were administered a short questionnaire, the General Health Questionnaire (GHQ-12), which screens for psychological symptoms. The GP recorded the reasons for each consultation, and was interviewed at the end of each day about selected patients to determine their opinion about the type of psychological problems experienced. Selected patients were then visited in their own homes and an extensive interview conducted, which included the Composite International Diagnostic Interview (CIDI) to determine mental health status, the World Health Organization's Disability Assessment Schedule (WHODAS) to determine disability, and a detailed exploration of use of health services. In the second phase of the study, patients were contacted by telephone at three, six, nine and 12 months, and both patients and GPs were re interviewed at 12 months. RESULTS: The study achieved a very high response rate among the GPs (90%). Nearly all eligible patients (93%) completed the GHQ screening, and their response rate was 70% for the first-phase interview. GPs thought that 54% of female and 46% of male patients had experienced some level of psychological problems in the past year. GHQ screening also found that more than half of those attending their general practitioner experienced some psychological symptoms at initial screening, and the CIDI interview found that more than one in three had a diagnosable mental disorder during the past 12 months. The most common mental disorders were depressive, anxiety and substance use disorders. These disorders were more common among younger than older general practice attenders, and comorbidity was high. CONCLUSIONS: Mental health problems are very common among general practice attenders. Contrary to the prevailing view that general practitioners seldom identify psychological problems in their patients, they identified about half their patients as having some type of psychological problems in the past year, although they considered that these were moderate or severe in about only one in ten patients. Further work from this large New Zealand study will focus on the nature of the relationship between disorder and disability, and on the recognition, management and outcome of psychological problems. PMID- 12740630 TI - A health profile of New Zealand youth who attend secondary school. AB - AIM: To determine the prevalence of selected health behaviours and protective factors in a representative population of New Zealand youth who attend secondary school. METHODS: The study sample comprised 12 934 Year 9 to 13 youth from 133 randomly selected secondary schools across New Zealand in 2001. A cross sectional, anonymous, self-report survey was conducted, incorporating 523 questions in a multimedia computer assisted self-interview (M-CASI) format. RESULTS: The school response rate was 85.7% and the student response rate was 75.0%, resulting in an overall response rate of 64.3%. The final dataset comprised 9570 students (males 46.2%, females 53.8%) belonging to diverse ethnic groups (Maori 24.7%, NZ European 55.3%, Pacific 8.2%, and Asian 7.2%). Most students (males 94.2%, females 90.3%) rate their health as good or better, and 90% report the presence of a caring adult in their family or at school. More than one quarter of students (males 27.2%, females 27.6%) report riding in a car driven by a potentially intoxicated driver within the last four weeks. Students report high levels of suicidal thoughts (males 16.9%, females 29.2%), suicide attempts (males 4.7%, females 10.6%), and depressive symptoms (males 8.9%, females 18.3%). CONCLUSIONS: This survey finds that most school students are healthy, but there are areas of serious concern including driving behaviours and mental health. Students report a high prevalence of positive connections with family and school; these connections are known sources of resiliency in the lives of young people. Findings of the current study support the implementation of the New Zealand Government's newly released youth policies: the Youth Development Strategy Aotearoa and the Youth Health Action Plan. PMID- 12740631 TI - Quality of life before and after heart, lung and liver transplantation. AB - AIMS: To compare the quality of life of heart, lung and liver transplant patients in New Zealand before and after transplantation and in comparison with New Zealand normative data for the general population. METHODS: All surviving transplant recipients and patients on the waiting list for a transplant from the Heart and Lung Transplant Unit at Green Lane Hospital and the New Zealand Liver Transplant Unit at Auckland Hospital were invited to participate in this study. 72% of the pre-transplant patients and 75% of the post-transplant patients completed a questionnaire, which included a standardised measure of quality of life - the MOS Short-Form 36 Health Survey (SF-36). RESULTS: Post-transplant patients reported significantly higher levels of physical and psychological functioning compared with pre-transplant patients. When compared with New Zealand normative data, post-transplant patients had poorer physical functioning but higher levels of psychological wellbeing and vitality. Time since transplant and the type of organ transplant were generally unrelated to quality of life. CONCLUSIONS: These data suggest that patients experience a dramatic improvement in quality of life following transplant, irrespective of the nature of the transplanted organ. These gains appear to be relatively independent of time since transplant. PMID- 12740632 TI - Into the unknown: the anticipation and experience of membership of independent practitioner associations. AB - AIMS: To understand why general practitioners (GPs) joined independent practitioner associations (IPAs), their concerns on joining, and the extent to which both positive and negative expectations have been realised. METHODS: A self complete postal questionnaire to a sample of IPA rank-and-file members invited their views on their decision to join, their satisfaction with leadership, and the experience of being in an IPA. RESULTS: The most popular reasons for joining were related to the uncertainties of the health sector environment, including the prospect of contracting and the place of general practice within the health sector. Aspirations on joining were largely realised, although at a general rather than specific level. Concerns over joining related mostly to day-to-day operation and practitioner autonomy, but were less strongly held and less likely than aspirations to be realised. Satisfaction with IPA leadership was quite high and associated with practitioner involvement in IPA activities. CONCLUSIONS: Results are consistent with the international literature, with importance attached by practitioners to both 'personal' and 'system' level aspirations. Research also suggests that where management remains 'connected' with rank-and file clinicians then perceived threats to autonomy are likely to be minimised. In moving towards primary health organisations, care needs to be taken not to undermine this 'connectedness' and therefore pose risks to the effective management of primary care. PMID- 12740634 TI - Minocycline-induced lupus: a case series. PMID- 12740633 TI - Ecstasy use in New Zealand: findings from the 1998 and 2001 National Drug Surveys. AB - AIMS: To examine changes in the use of ecstasy, current conditions of supply, harms resulting from use, and the demographics of users. METHODS: National Drug Surveys were conducted in 1998 and 2001. In each survey, a representative national sample of approximately 5500 people aged 15-45 years were asked about their drug use, including ecstasy use, using a Computer Assisted Telephone Interview (CATI) system. Response rates of 79% and 80% respectively were achieved. RESULTS: Last-year use of ecstasy increased from 1.5% in 1998, to 3.4% in 2001. Large increases were found among men aged 20-24 (4.3% to 12.5%), and 25 29 (3.2% to 8.8%). In 2001, 43% of users thought ecstasy was easier to obtain and 29% thought the price was lower compared with a year earlier. About one in ten ecstasy users reported problems related to 'energy and vitality', 'financial position', 'health', and 'outlook on life'. Ecstasy users were predominantly male, aged 20-29, European and single, but were from a broad range of occupational and income-earning groups. CONCLUSIONS: The use of ecstasy in New Zealand increased between 1998 and 2001. Conditions of supply became easier. Users reported problems related to use in a range of areas of their lives. There was little evidence to suggest ecstasy use was limited to high-income-earning professionals. PMID- 12740635 TI - An uncommon clinical presentation of asbestos-related disease. PMID- 12740636 TI - Cyclosporin successfully treats red cell aplasia associated with myelodysplasia. PMID- 12740637 TI - The lost generation. PMID- 12740638 TI - Attention-deficit/hyperactivity disorder. PMID- 12740639 TI - HbA1c standardisation issues: should New Zealand follow the DCCT or the IFCC position? PMID- 12740640 TI - Characterization of interleukin 10 and NO release by transplantable melanoma cell lines with regard to their progression. AB - The results from our investigations concerning the secretory activity of interleukin 10 (IL-10) by two transplantable melanoma cells showed that a spontaneous alteration of the native-melanotic line into an amelanotic form and the tumor progression connected with it, were accompanied by a 6-fold decrease of the IL-10 content in the supernatant of these melanoma line cell culture. Simultaneously, the intracellular content of IL-10 indicated that there was only a small population of IL-10 positive cells, numerically similar in both melanoma cell lines. A comparison of the IL-10 and nitric oxide (NO) secreted by both melanoma lines does not show any correlation between the changes in the content of these substances, which seems to indicate that NO does not act as an autocrine regulator of IL-10 secretion. PMID- 12740642 TI - Expression of p65, DD3 and c-erbB2 genes in prostate cancer. AB - The expression of p65, DD3 and c-erbB2 genes was analyzed in 39 histologically verified human prostate cancers. The expression of p65 and DD3 genes was observed in significant percentage in well- and moderately-differentiated tumors. Both genes expression was lower in poorly differentiated tumors. On the contrary, c erbB2 gene expression increased with advanced histological grading and reached the highest percentage in poorly-differentiated cancers. In the all investigated groups straight dependence between p65 and DD3 genes expression occurred. Opposite dependence was noticed in expression of p65/DD3 and c-erbB2 genes. PMID- 12740641 TI - Modulation of HLA class I expression in multidrug-resistant human rhabdomyosarcoma cells. AB - An abnormal HLA expression has been detected in some tumors including rhabdomyosarcoma (RMS). Classical cytotoxic treatment of these tumors, the most common childhood soft tissue malignancy, may induce multidrug resistance (MDR) associated with the expression of a 170-kDa membrane-associated glycoprotein (P glycoprotein). In order to analyse the connection between modulation of HLA expression and the development of the MDR phenotype mediated by P-glycoprotein in RMS, we used three resistant RMS cell lines; two of these resistant cell lines (TE.32.7.DAC and RD-DAC) were established by in vitro exposure to actinomycin D, a drug of choice in the treatment of RMS; the resistant RMS- GR cell line was established from an embryonal RMS tumor after polychemotherapy. Our results showed that all the resistant cell lines showed a significant increase in the expression of HLA class I surface antigens in comparison to drug-sensitive cells. Blockade of P-glycoprotein with verapamil led to a decrease in HLA class I expression in RMS resistant cell lines. However, no modulation of HLA class II expression was observed in any of the three analyzed cell lines. These findings support the hypothesis that the development of resistance mediated by mdr 1/P glycoprotein, directly influences the expression of HLA class I in RMS cells, inducing to upregulation. This effect may be relevant to the application in RMS of immunotherapy against tumor-associated antigens presented by HLA class I molecules. PMID- 12740643 TI - Lipoxygenase inhibitors suppress proliferation of G5:113 fibrosarcoma cells in vitro but they have no anticancer activity in vivo. AB - Nordihydroguaiaretic acid (NDGA) and esculetin, both nonspecific inhibitors of lipoxygenases (LOX), were found to suppress expressively the in vitro proliferation of fibrosarcoma cells G5:113 in concentrations ranging from 10 to 50 microM. Subsequent flow-cytometric analysis of the cell cycle showed that both these drugs significantly decreased the percentage proportion of cells in the G0/G1-phase and simultaneously increased significantly this proportion in the S phase. No apoptosis was detected in the whole range of concentrations studied, from 2.5 to 50 mM. On the contrary, in experiments in vivo, neither NDGA nor esculetin had any curative effect if they were repeatedly injected intraperitoneally (i.p.) into mice bearing tumors growing from subcutaneously (s.c.) transplanted G5:113 cells. Pretreatment of the fibrosarcoma cells with NDGA or esculetin in vitro preceding their s.c. transplantation into mice did not result in suppression of the tumor growth, either. Finally, if G5:113 cells were injected intravenously and the mice were subsequently treated repeatedly with i.p. injections of NDGA, decreased survival and increased number of surface lung metastases were observed in the NDGA-treated group. Thus the suppressive action of inhibitors of LOX on the growth of fibrosarcoma cells in vitro was not reflected in their anti-tumor effects in vivo. PMID- 12740644 TI - Comparison of the carcinogenic potential of streptozotocin by polarography and alkaline elution. AB - The carcinogenic potential of streptozotocin (STZ) was evaluated by the polarographic determination of its reduction potential in the presence of alpha lipoic acid and detection of DNA single-strand brakes by alkaline elution. After STZ electrochemical reduction in an anhydrous solvent, the half-wave potential (E1/2 ) was determined to be -1.340 V. The parameter of the carcinogenic potential (tg alpha) for STZ was 0.400. This is in good agreement with WHO data regarding STZ carcinogenicity. Additionally, it is in the good agreement with the tg alpha value determined for the positive control used, N-nitroso-N-methylurea (NMU), which was found to be 0.459. The 3 hours exposure of A549 human lung tumor cells to 250, 500, and 1000 nmol/ml STZ was followed by DNA single-strand brakes detection using the alkaline elution method. NMU, the positive control, was tested under identical experimental conditions at the same concentrations. Without metabolic activation, NMU induced a significant formation of DNA single strand brakes only at 1000 nmol/ml. In the presence of the metabolic activation, NMU caused a significant, concentration-dependent formation of DNA single-strand brakes. In the absence of metabolic activation, STZ induced no significant formation of DNA single-strand brakes at any concentration used. In the presence of metabolic activation, STZ caused a significant, concentration-dependent formation of DNA single-strand brakes. The results of this study underline the crucial role of using a metabolic activation system when carcinogenic potential of drugs and chemicals is investigated in vitro studies. Results of polarographic experiments and alkaline elution correlate well with each other and they indicate that these methods are useful to early predict the carcinogenic potential of STZ and other xenobiotics. PMID- 12740645 TI - Usefulness of technetium-99m methoxyisobutylisonitrile liver single photon emission computed tomography to detect hepatocellular carcinoma. AB - Technetium-99m methoxyisobutylisonitrile (Tc-99m MIBI) has been shown to be useful in identifying several types of tumors, such as breast, lung and thyroid cancers. The usefulness of Tc-99m MIBI liver imaging in detecting hepatocellular carcinoma (HCC) is still controversial. In this study, 22 patients with HCC performed Tc-99m MIBI liver single photon emission computed tomography (SPECT). Twenty of 22 patients (90.9%) showed negative liver SPECT findings without significant Tc-99m MIBI uptake in HCC, and only 2 patients (9.1%) showed positive liver SPECT findings with significant Tc-99m MIBI uptake in HCC. In addition, no significant correlation between liver SPECT findings with sex, age, alpha feto protein serum level, HCC differentiation, and virus hepatitis status was found. We concluded that Tc-99m MIBI liver SPECT is not a sensitive tool to detect HCC. PMID- 12740646 TI - Rapid HPLC analysis of melphalan applied to hyperthermic isolation limb perfusion. AB - Hyperthermic isolated limb perfusion (HILP) with melphalan (MH) as a standard cytotoxic drug has been performed in 28 patients suffering from malignant melanoma. MH has been administered by HILP via extracorporeal circulation system. The drug given locoregionally reduces subsequent toxicity of organs. For all that residues can leak into the systemic circulation during HILP. Because of known carcinogenic potential and secondary cancer formation, the main interest of this work is to determine MH concentration profile in the patient plasma during and after HILP and evaluation of its potential toxicity in patients. Reversed-phase HPLC assay, which uses isocratic elution and fluorimetric detection has been shown to be sensitive, reliable and suitable for routine analyses. The assay was validated for the concentration range of 50-2500 ng.ml-1 with the limit of detection (LOD) 6.881 ng.ml-1. The samples were treated by methanol precipitation with the recovery more than 80%. The stability of standard solutions and methanolic extracts of MH were also followed. The concentration profile of MH in patient samples has been pursued in three time points during and after chemoperfusion (45 min after application of MH in extracorporeal circulation, 10 min after the joining the extremity to systemic circulation and one hour after the great vessels reconstruction). The concentrations of MH ranged 100-1500 ng.ml 1 and varied from patient-to-patient. Some complications were observed after HILP in 11 patients and are correlated with the higher con- centrations of MH (over 150 ng x ml-1) found in plasma. PMID- 12740647 TI - Thermoluminescent dosimetry in rotary-dual technique of the total skin electron irradiation. AB - The aim of the study was to discuss the results of thermoluminescent dosimetry (TLD) in rotary-dual technique of the total skin electron irradiation (TSEI RD), to confirm beam calibration and monitor unit calculations and to provide data for making clinical decisions. Between May 2001 and April 2002, in 3 cases of mycosis fungoides, 736 dosimetric checks were performed in 34 points at the skin. CaF2:MnTLD-400 cubes (1/8"x1/8"x0.015") were used for in vivo dosimetry. Doses were computed and analyzed for all locations. Percent of described dose and SD for the following localizations from 34 points were: anterior abdomen (reference point) 100+/-6%, upper back 100+/-8%, right calf 98+/-10%, left foot (mid dorsum) 97+/-8%, posterior neck 93+/-6%, right hand (mid dorsum) 78+/-10%, hand fingers 57+/-10%, top of right shoulder 56+/-14%, left groin 35+/-20%, perineum 22+/-17%. The correlations between patient's height and measured doses were sufficient for the following localizations: scalp (top rear), occiput, elbows, hand fingers and hands (mid dorsum). The correlations between obesity index and measured doses were sufficient for the following localizations: shoulders and lateral neck, groins, and perineum. Dosimetric checks at the reference point confirm that our beam calibration technique and monitor unit calculation are accurate. TLD shows that for some parts of the skin such as shoulder, hands and perineum boost fields were required. The correlations with obesity index and height for several sites suggest that boost fields must be customized for each patient. PMID- 12740648 TI - Immunocytochemical detection of p21ras, Raf-1, ERK1/MAP kinase and PKC isoforms in a 20-methylcholanthrene-induced transformed murine embryonal fibroblast cells in culture. AB - An immunocytochemical study using antibodies against p21ras, Raf-1, MAP kinase/ERK1 and PKCalpha, beta, gamma, delta, epsilon, zeta, isoforms were performed on a 20-methylcholanthrene-induced transformed murine embryonal fibroblast cells in both in vitro and in vivo growth conditions. Altered expression of p21ras, Raf-1, MAP kinase in this particular cell line strongly supported the previous findings of the activation of one component of signal transduction under the influence of the other in the MAP kinase cascade of signal transduction during neoplastic transformation and which also seemed to be involved in CNCI-PM-20 cell line. The altered expression of PKCalpha, beta, and delta was thought to be an epigenetic event occurring under the indirect influence of other changes in these cells. Host physiology and metabolism did not have much impact on the expression of these gene products after biological incubation of these cells in syngenic host. PMID- 12740649 TI - Correlation of proliferating cell nuclear antigen and bcl-2 expression with tumor front grading and metastasis in laryngeal squamous cell carcinoma. AB - This study was designed to examine the immunohistochemical expression of proliferating cell nuclear antigen (PCNA) and bcl-2 protein in 45 cases with advanced laryngeal squamous cell carcinoma who had undergone total laryngectomy with unilateral modified radical neck dissection, and the relation of this expression to some prognostic factors such as tumor front grading and neck lymph node metastases. Sections were reevaluated for routine histologic grade, tumor front grading and neck lymph node metastases, and were stained with monoclonal antibodies against PCNA and bcl-2. Significant correlation was present between the severity of PCNA expression and incidence of lymph node metastasis (p<0.05). No correlation was found between the severity of PCNA expression and tumor front grading. Bcl-2 expression did not associate with either parameters. In conclusion, PCNA is important in predicting prognosis and no association is present between the bcl-2 protein expression and prognostic factors. PMID- 12740650 TI - Fine needle aspiration biopsy in a diagnostic workup algorithm of salivary gland tumors. AB - The ultrasonic diagnosis of salivary gland diseases can give a more accurate information than clinical data alone. In diagnosis of the parotid gland diseases, it is necessary to differentiate clearly tumors from other diseases. When a tumor is suspected, its presence should be confirmed, and determined if it is solitary, multiple, unilateral or bilateral, intra or extraglandular, and benign or malignant. In the period of May 1998 to January 2001 ultrasound examinations were performed in 354 patients, FNAB with ultrasound control were performed in 152 patients with a mean age of 49.7 years. The ultrasound and cytologic findings were correlated with surgical and histologic findings. Results indicate that in patients with solid mass of the salivary gland, ultrasound examination confirmed the finding of salivary gland tumor in all patients (100%). In non-solid diseases of salivary glands the diagnostic accuracy was slightly lower. The use of ultrasound techniques in the study of salivary gland pathology is well justified, due to its capacity to provide high resolution and improving clinical diagnosis. FNAB is a simple quick, accurate and virtually complications free investigative modality. The data on its ability to distinguish between lesions requiring surgery or not are encouraging. PMID- 12740651 TI - Inhibition of adhesion breast cancer cells by anticoagulant drugs and cimetidine. AB - Recent studies suggest that anticoagulant drugs and cimetidine therapy in malignancy may improve cancer survival and inhibit the metastatic process. In this study we investigated and compared the effects of anticoagulant drugs (unfractionated heparin, warfarin, acetylsalicylic acid, low-molecular-weight heparins--nandroparinum, enoxaparinum, dalteparinum and reviparinum), cimetidine and combination of cimetidine with anticoagulants on adhesion of highly invasive breast cancer cells lines - BT 549 and MDA-MB-231 (MDA 231)--in vitro. High antiadhesion effect was observed with cimetidine, warfarin and acetylsalicylic acid. Low-molecular-weight heparins had a small antiadhesion effect in independent use. In combination with cimetidine, a potential effect of cimetidine on the antiadhesion was observed. The antiadhesion effect was dependent on the type of the cancer cell line. Different effects between cell lines BT 549 and MDA 231 were observed. The strongest antiadhesion effect was obtained using the combination of cimetidine with acetylsalicylic acid. In the majority of applications of the drugs and their combinations, a proportional antiadhesion effect was dependent on the concentration and time. We suppose that anticoagulant drugs might have higher antimetastatic effect in combination with cimetidine. The choice of anticoagulants can decrease the adhesion, decrease tumor angiogenesis and cause the shortening of blood clotting time. Cimetidine can decrease the adhesion of cancer cells and increase the activity of NK cells. Indeed, according to our results, application of cimetidine and anticoagulant drugs intensifies the antiadhesion effect together with other antimetastatic effects. PMID- 12740652 TI - Conservative treatment for carcinoma of the anus--a report of 35 patients. AB - Conservative treatment for carcinoma of the anus has become the standard care for this malignancy. In this study we report on our experience with this method with particular emphasis on treatment outcome and acute toxicity. Between April 1991 and February 2002, 35 patients (male/female ratio 0.35) with UICC T(1-i) N(0-3) M(0) squamous cell carcinoma of the anal canal or anal margin were treated with chemo-radiation (31 patients) or radiotherapy alone (4 patients). Three patients had previously undergone local tumor excision with anus preservation. The total tumor dose of 48 to 60 Gy was delivered either by split-course or continuous radiation therapy to the pelvis, followed by a local boost to the primary tumor. Chemotherapy included one or two cycles of mitomycin C (10-15 mg/m(2) day 1) and 5-fluorouracil (450-750 mg/m(2) day 1 to 4 or 5) given during the first and the last part of irradiation. Complete tumor remission was obtained in 26 (76%) out of 34 evaluable patients. Clinically persistent disease was found in five (17%) and three (7%) patients treated with chemo- radiation and radiation alone, respectively. In four of these cases salvage surgery was performed. With a median follow-up of 49 months (range 2-131 months) local recurrence occurred in four patients (12%), and distant metastases - in two (6%). Overall, local treatment failure was observed in twelve patients (35%) including eight with T3 and one with T4 tumor. Local control was maintained until the last follow-up or death in 22 patients (65%). An actuarial 5-year overall and colostomy-free survival rates were 63% (CI, 45-81%) and 45% (CI, 25-64%), respectively. Nineteen patients (54%) experienced acute toxicity, predominantly hematologic and gastrointestinal, and severe effects including one death occurred in 11 patients (31%). Late sequelae including chronic diarrhea, edema of genitalia and legs, impaired sexual activity, and bone fractures were observed in eight patients (24%). Moderate anal stool incontinence occurred in three patients (9%). In conclusion, conservative management of anal carcinoma allows durable colostomy-free survival in a proportion of patients. However, the risk of local failure is relatively high in patients with large primary tumors. Combined chemo-radiation is associated with relatively high rate of acute toxicity. PMID- 12740653 TI - Giant ganglion cyst of the quadriceps femoris tendon. AB - We present the case of a ganglion cyst with 11 x 3 x 2 cm dimensions originating from the quadriceps femoris tendon. The patient presented with palpable mass and chronic pain in the anterolateral thigh region. The pain increased with walking. Ultrasonography and computed tomography revealed a cystic, well defined lesion in the vastus lateralis muscle. The patient was operated on and the cyst excised; it was seen to be a ganglion cyst. The patient is asymptomatic 6 months after surgery. This is a very rare localization for a giant synovial cyst, and preoperative diagnosis is a challenge for the surgeon. PMID- 12740654 TI - [Present status of treatment in nasopharyngeal carcinoma?]. AB - AIM: Review of the evolution of combined treatment strategies in nasopharyngeal carcinoma. RESULTS: Radiotherapy is accepted standard for treatment of nasopharyngeal cancer. Nevertheless, there is no uniform opinion with regard to doses, fractionation, technique or use of systemic chemotherapy. It is hardly possible to compare the results of recent and historical trials because of different staging systems and because nasopharyngeal cancer occurring in the Oceano-Asian region are biologically different to those in Western countries. Conclusions drawn from former, mostly retrospective analyses are not applicable to newer standards regarding the developments in diagnostics and therapy. Presently simultaneous chemoradiotherapy is standard for lymph node positive nasopharyngeal cancer. CONCLUSIONS: It will be necessary to treat patients with different histologic subtypes with an uniform treatment schedule to define the place of combined modality treatment. This will probably be the only way to develop treatment concepts for distinct stages and biological entities. PMID- 12740655 TI - Usefulness of tumor volumetry as a prognostic factor of survival in head and neck cancer. AB - BACKGROUND: The TNM classification system of tumor stage does not always reflect the actual tumor mass present at diagnosis. This study aimed at evaluating the prognostic value of volumetric data regarding survival in head and neck cancer patients being treated with either cisplatin or carboplatin administered concomitantly with radiotherapy. PATIENTS AND METHODS: We retrospectively analyzed 107 patients suffering from squamous cell carcinoma of the head and neck in a Greek-German cooperative study (see Table 1). All patients were treated by radiotherapy and concomitant chemotherapy. 65 patients received chemotherapy with carboplatin and 42 with cisplatin. More than 6,200 CT scans were analyzed by digitalization of contours which subsequently led to the computation of the tumor volume (primary and macroscopic lymph node metastases). RESULTS: Median follow-up was 43 months and median survival 30 months. Median initial tumor volume was 32.5 ml (range 2.1-220.1 ml) in the carboplatin and 44.4 ml (range 3.2-202.5 ml) in the cisplatin group (see Figure 1). After treatment, tumor volumes did not differ significantly (median of 3.1 ml [range 0.0-167.1 ml] and 3.5 ml [range 0.0-166.0 ml], respectively). 41 patients (63.1%) died in the carboplatin group and 22 patients (52.4%) in the cisplatin group (see Figure 2). Pretherapeutic tumor volume was prognostic with respect to survival while TNM classification and age were not. Pretherapeutic tumor volume was negatively and percent decrease in tumor volume positively associated with survival (see Tables 2 and 3). CONCLUSION: Knowledge of the initial tumor volume adds valuable information in terms of prognosis. Initial tumor volume should be included in all future clinical trials regarding head and neck cancer patients. PMID- 12740656 TI - Reirradiation of locally recurrent nasopharyngeal carcinoma. AB - PURPOSE: To study the efficacy of reirradiation as salvage treatment in patients with locally recurrent nasopharyngeal carcinoma. PATIENTS AND METHODS: Between 1993 and 2000, 20 consecutive patients (twelve males and eight females) with nasopharyngeal cancer, previously irradiated in different Hungarian institutions, were reirradiated for biopsy-proven locally recurrent tumor. Histologically, 85% of the patients had WHO type III, 5% type II, and 10% type I disease. Stages I-IV (AJCC 1997 staging system) were assigned to five (25%), seven (35%), five (25%), and three (15%) patients, respectively; none of them had distant metastases, and only eight (40%) displayed regional dissemination. The median time period between termination of primary treatment and local recurrence was 30 (range, 10-204) months. Brachytherapy was the method most frequently used: in ten cases alone (especially for rT1 tumors), and in eight cases in combination with external beam therapy. Two patients with locally advanced disease underwent external beam therapy only. The median dose in the event of brachytherapy alone was 20 Gy (4 x 5 Gy or 5 x 4 Gy, range, 16-36 Gy), and the dose range for exclusive external irradiation was 30-40 Gy. In cases of combined irradiation, a median 20-Gy brachytherapy (range, 16-40 Gy) was associated with 30-40 Gy of external irradiation. Radiotherapy was supplemented by neck dissection (six patients), nasopharyngectomy (one patient), or chemotherapy (eleven patients). RESULTS: 16 patients were reirradiated once, three twice, and one patient three times, with a median equivalent dose for tumor effect of 36 Gy (mean, 44 Gy; range, 19-117 Gy; the estimated alpha/beta-ratio was 10 Gy). The median equivalent dose of reirradiation for late effect on normal tissue (with an estimated 70% delivery of the tumor dose) amounted to 30 Gy (mean, 37 Gy; range, 13-101 Gy, estimated alpha/beta-ratio 3 Gy). After a median follow-up of 37 (range, 12-72) months, the overall survival was 60% (12/20). Seven of the twelve surviving patients are currently tumor-free. After primary irradiation, xerostomy occurred in all patients as an unavoidable side effect of treatment. Following reirradiation, a severe (grade 3 or higher) late toxicity (CTC criteria, version 2) has been observed in two tumor-free patients (10%) so far (necrosis of soft palate and paresis of glossopharyngeal nerve). CONCLUSION: Retreatment of nasopharyngeal carcinoma with radiotherapy (preferably a combined modality), can result in longterm local control and survival in a substantial proportion of patients, at the price of an acceptable morbidity. PMID- 12740657 TI - Remission rates in breast cancer treated with preoperative chemotherapy and radiotherapy. AB - PURPOSE: Evaluation of remission rates after neoadjuvant chemotherapy alone or followed by preoperative radiotherapy. PATIENTS AND METHODS: 194 women with 198 biopsy-proven breast tumors were evaluated in this retrospective study. Of the 198 cases evaluated, 64 received neoadjuvant chemotherapy followed by surgery and adjuvant irradiation (CT group). In 134 cases, sequential preoperative chemo /radiotherapy (CT-RT group) was given. In both groups, endocrine treatment was initiated in case of positive hormone receptor status after chemotherapy. The whole breast was homogeneously irradiated using 2-Gy fractions up to a total dose of 50 Gy, followed by a boost of 6-11 Gy to the tumor. RESULTS: A histologically proven complete remission (pCR) was achieved in 3% (2/64) in the CT and in 42% (56/134) in the CTRT group. The logistic regression analysis, including clinical tumor category (cT), lymph node (cN) and metastasis status (cM), grading (G), hormone receptor status (HRS), number of preoperative chemotherapy cycles, preoperative tumor volume, and preoperative radiotherapy, revealed that HRS (p = 0.0232) and radiotherapy (p < 0.0001) were significant factors for achieving pCR. CONCLUSION: Combination of neoadjuvant chemo-/radiotherapy results in significantly higher rates of complete remission than neoadjuvant chemotherapy alone. The significance for tumor-free and overall survival has to be evaluated. PMID- 12740658 TI - Dosimetric assessment of the field abutment region in head and neck treatments using a multileaf collimator. AB - BACKGROUND AND PURPOSE: The use of conventional asymmetric collimators for junctioning of abutted fields can lead to significant dose inhomogeneity, due to jaw misalignment. However, recent technologic advances enable us to fabricate much finer leafpositioning accuracy. Consequently, it is anticipated that the use of multileaf collimator (MLC) will potentially improve dose homogeneity at the junction of abutted fields. In this work, we evaluated the dose inhomogeneities at the match-plane in monoisocentric three-field head and neck setups, using MLC for field abutment. MATERIAL AND METHODS: To define either the anterior or the lateral fields, the MLC was used with either the longitudinal (0 degree angle) or the transverse (90 degree angle) settings. For 0 degree setting, each leaf moves in a direction perpendicular to the gantry rotation axis, hence the "tongue and groove" (T&G) design can effect matching-area dose at the side of the leaf (Figure 1a). For 90 degree setting, the rounded shape of the leaf produces its effect at the leaf end (Figure 1b). Four combinations of abutted anterior field and abutted lateral field defined by MLC, i.e., abutted using MLC side-by-side, side-by-end, end-by-side and end-by-end, were compared (Table 1). Dose inhomogeneity was measured at the junction of the two abutted fields with films in a solid water phantom (Figure 2). The effect of jaw settings as a backup diaphragm on the dose distribution was also studied. Reproducibility of the results was confirmed by repeated measurements over a 1-year period. RESULTS: Abutted fields using MLC side-by-side caused underdose of approximately 15% (Figure 3a). Abutted fields using MLC side-by-end produced > 10% overdose that could be improved to +/- 1% for 0.5 mm overlap of the leaf end from the lateral portals (Figure 4). When using end-by-side, an overdose of approximately 15% was observed. However, the dose improved to a homogeneous dose for 0.8 mm overlap of leaf end from the anterior portal (Figure 5). End-by-end showed an overdose of > 20% (Figure 3b). This overdose could be smoothed out by overlaps of both leaf ends by 0.8 mm from both lateral and anterior portals (Figure 6). The ideal jaw position was found to be at 1 mm away from the beam central axis in any combination (Table 2). CONCLUSION: The use of MLCs for photon field junction matching is appropriate and represents an alternative approach to the problem of field matching using the asymmetric jaws in head and neck treatments. PMID- 12740659 TI - Acute side effects during 3-D-planned conformal radiotherapy of prostate cancer. Differences between patient's self-reported questionnaire and the corresponding doctor's report. AB - BACKGROUND: Radiotherapy-induced side effects are often scored retrospectively according to the EORTC/RTOG scores for organs at risk by reviewing the medical records. Some studies could prove an over- or underestimation of side effects as assessed by the medical professionals. The aim of this study was to prospectively evaluate differences in side effects as described by the doctors and the patients. PATIENTS AND METHODS: 47 patients with prostate cancer were questioned about their side effects by a radiotherapist and asked to fill in a questionnaire at the start, in the middle and at the end of radiotherapy. The data of this questionnaire and the doctor's report were scored according to the German version of the EORTC/RTOG scores for gastrointestinal (GI) and genitourinary (GU) side effects and subsequently compared. We distinguished between "moderate" disagreement (better/worse by one grade, assessed by the doctor) and "pronounced" disagreement (better/worse by two grades, assessed by the doctor). RESULTS: The number of GI and GU side effects increased during radiotherapy both according to data obtained from the doctor and the patient questionnaire. Comparing doctors' reports with patients' questionnaires, for GI side effects an agreement was found in 22/47 patients, "moderately better" scores by the doctor's report were found in 13/47 patients, and "moderately worse" scores in 9/47 patients on average. "Pronouncedly better and worse" scores were found in 2/47 patients. For GU side effects an agreement was seen in 22/47 patients, "moderately better" scores in 17/47 patients and "moderately worse" scores in 3/47 patients. Regarding GU side effects, only pronouncedly better scores, as assessed by the doctor, were found in a mean of 4/47 patients. If the EORTC/RTOG score is used in its original English version, a difference is found, particularly in the assessment of GU side effects, resulting in an higher amount of agreement concerning GU side effects and a minor amount of "pronounced disagreement". CONCLUSION: In order to evaluate radiation-induced side effects, a patient's self-reported questionnaire should be included in the analysis of morbidity, above all for grade 0, 1, and 2 side effects. The validity of data seems to be questionable, particularly in the assessment of grade 0, 1 and 2 side effects, if only data from the doctors' reports are taken into account. The German version of the EORTC/RTOG score--not including the pretreatment status--leads to different results, particularly in the assessment of grade 0, 1, and 2 urinary side effects, which asks for a revision. PMID- 12740661 TI - Cell adhesion-mediated radioresistance (CAM-RR). Extracellular matrix-dependent improvement of cell survival in human tumor and normal cells in vitro. AB - BACKGROUND: Cell-extracellular matrix (ECM) contact is thought to have great impact on cellular mechanisms resulting in increased cell survival upon exposure to ionizing radiation. Several human tumor cell lines and normal human fibroblastic cell strains of different origin, all of them expressing the wide spread and important integrin subunit beta1, were irradiated, and clonogenic cell survival, beta 1-integrin cell surface expression, and adhesive functionality were investigated. MATERIAL AND METHODS: Human tumor cell lines A172 (glioblastoma), PATU8902 (pancreas carcinoma), SKMES1 (lung carcinoma), A549 (lung carcinoma), and IPC298 (melanoma) as well as normal human skin (HSF1) and lung fibroblasts (CCD32) and human keratinocytes (HaCaT) were irradiated with 0-8 Gy. Besides colony formation assays, beta 1-integrin cell surface expression by flow cytometry and adhesive functionality by adhesion assays were analyzed. RESULTS: All cell lines showed improved clonogenic survival after irradiation in the presence of fibronectin as compared to plastic. Irradiated cells exhibited a significant, dose-dependent increase in beta 1-integrin cell surface expression following irradiation. As a parameter of the adhesive functionality of the beta 1 integrin, a radiation-dependent elevation of cell adhesion to fibronectin in comparison with adhesion to plastic was demonstrated. CONCLUSION: The in vitro cellular radiosensitivity is highly influenced by fibronectin according to the phenomenon of cell adhesion-mediated radioresistance. Additionally, our emerging data question the results of former and current in vitro cytotoxicity studies performed in the absence of an ECM. These findings might also be important for the understanding of malignant transformation, anchorage-independent cell growth, optimization of radiotherapeutic regimes and the prevention of normal tissue side effects on the basis of experimental radiobiological data. PMID- 12740660 TI - [Concurrent chemotherapy and radiation therapy for unresectable locally advanced carcinoma of the esophagus. Phase II study and clinical review on literature]. AB - BACKGROUND: Neither surgical advances nor those in therapeutic radiology have been able to significantly reduce the mortality related to esophageal carcinomas. The results of combining: first surgery, then radiation therapy, which have been unsatisfactory for decades, encourage therapeutic concepts involving a variety of modalities. PATIENTS AND METHODS: For 50 patients with unresectable locally advanced esophageal carcinomas, a palliative concurrent chemotherapy and radiation therapy was carried out according to the "intent to treat" principle. The aim was a minimal dose of 40 Gy. The concurrent chemotherapy was carried out using cisplatin/5-FU during the 1st and 4th weeks of radiation therapy. In the case of partial or complete remission, the chemotherapy was to be continued as maintenance therapy with a maximum of four cycles. In the case of no change or minor response, instead of maintenance chemotherapy, the dose of local radiation was to be increased by means of brachytherapy. RESULTS: The median survival rate for the entire population under study was 8.7 months. The survival rates of 1, 2, 3, 4, and 5 years were, respectively, 38%, 20.5%, 13.7%, 6.8%, and 6.8%. The remission rates were as follows: NC: 14 patients (28%), PR: 32 patients (64%), CR: 4 patients (8%). 17 patients (34%) tolerated the full concurrent chemotherapy; only twelve patients (24%) tolerated supportive chemotherapy. The following factors exhibited a significant correlation to survival: the intensity of the chemotherapy, the Karnofsky index, the age of the patients, and the improvement of oral food intake. CONCLUSIONS: The concurrent chemotherapy was toxic and the benefit to the patients questionable. At best, meta-analyses of randomized studies along the lines of "evidence-based medicine" demonstrate for concurrent chemotherapy and radiation therapy an improvement of 2-year survival rates, but with these also involving a high level of toxicity. Due to the heterogeneous data available, the value of the primary, sequential treatment combining chemotherapy and radiation therapy is uncertain. PMID- 12740662 TI - Optimization of radiation therapy for locally advanced adenoid cystic carcinomas with infiltration of the skull base using photon intensity-modulated radiation therapy (IMRT) and a carbon ion boost. AB - BACKGROUND: Tumor doses > 70 Gy are needed for local control in adenoid cystic carcinomas. These tumor doses cannot be delivered if the tolerance doses to neighboring organs at risk (OAR) are respected. This treatment planning study investigates the physical advantage of combined photon intensity-modulated radiation therapy (IMRT) plus carbon ion boost compared to photon IMRT alone. PATIENTS AND METHODS: For nine patients, treatment plans were generated using a) photon IMRT alone (integrated boost concept), and b) sum plans consisting of a photon IMRT plan and a carbon ion boost plan. 54 Gy were prescribed to the planning target volume 1 (PTV1), the boost volume (PTV2) received 72 Gy. The tolerance doses of the delineated OAR were strictly adhered to. Plan quality of IMRT plans and sum plans was compared using adequate physical parameters. RESULTS: Both therapy techniques lead to highly conformal dose distributions that allow the prescription of the desired target doses. Target conformality and heterogeneity as well as target coverage for PTV1 are comparable for both techniques. The target coverage for PTV2 can be significantly improved using carbon ion beams (median 95% coverage 93.7% vs 87%; p = 0.039). Furthermore, the mean doses to the OAR can be reduced by 8.3% (median % reduction of mean doses to OAR; p = 0.00001) using carbon ions. CONCLUSIONS: The combination of photon IMRT with carbon ions improves the target coverage for the boost volume and offers better sparing of OAR close to the PTV2 (gross tumor volume) in comparison with photon IMRT alone. A clinical study has been initiated to evaluate whether these potential advantages translate into clinical benefit. PMID- 12740666 TI - Two cases of cerebral aspergillosis successfully treated with voriconazole. AB - Described here are two cases of cerebral aspergillosis successfully treated with voriconazole after the failure of first-line treatment with amphotericin B deoxycholate. In both cases, clinical and radiological cure was achieved within 6 weeks. Voriconazole should be considered as first-line therapy for cerebral aspergillosis. PMID- 12740668 TI - Functional outcome and subset identification in RA patients from meridional Europe: analysis of a Spanish cohort. AB - The aim of this study was to study the short-term functional and anatomical prognosis of rheumatoid arthritis (RA) in a series of Spanish patients and to identify different subsets of patients as well as possible baseline factors associated with specific outcomes. All patients seen in our division who met the ACR criteria for RA and with disease duration between 2 and 7 years were eligible for the study. Available patients were further evaluated at the clinic for disease activity using biological tests and joint indices as joint counts and Thompson's index, functional capacity using the ACR functional classification (ACR-FC) and the modified Health Assessment Questionnaire (M-HAQ) and radiologic damage by the Sharp's radiologic scoring method. Cluster analysis was used to identify different clinical subsets of patients. One hundred and sixty-three patients were eligible for the study, 13 could not be located or refused to participate and 12 had died. Mean (+/-SD) age at disease onset and mean disease duration were, respectively, 56(+/-14) years and (55+/-20) months. Median (interquartile range) of M-HAQ was 0.4 (0.1-1.1) and 41% of patients were in III or IV ACR-FC. The majority of patients (93%) showed radiologic lesions and 65% had erosions. Cluster analysis identified three subsets: cluster I (70% of patients) was characterised by a good prognosis, cluster II (13%) by a high level of disease activity, and cluster III (17%) by a greater anatomic damage and longer disease duration. No baseline predictive markers were found for these different outcomes. We concluded that RA portends an overall poor short-term prognosis in a relative large percentage of our patients with significant anatomic and functional sequelae. Aggressive management is specially indicated in this subgroup of patients, although definitive prognostic markers for its early identification are still lacking. PMID- 12740669 TI - Doppler echocardiographic evaluation of ventricular function in patients with rheumatoid arthritis. AB - Cardiac involvement in rheumatoid arthritis (RA) has been reported previously. However, evaluation of ventricular function in this disease by the use of recently proposed Doppler echocardiographic methods has not been reported before. Thus, the aim of this study was to evaluate ventricular function by measurement of myocardial performance index (MPI) and transmitral flow propagation velocity (TFPV). Thirty-two patients with long-standing RA and 32 control subjects (mean ages 52 +/- 11 and 50 +/- 10 years, respectively) participated in this study. Systolic function was assessed by subjective evaluation of wall motion for both ventricles and by fractional shortening for the left ventricle (LV). LV diastolic function was evaluated by standard pulsed-wave Doppler echocardiography, MPI and TFPV. Right ventricular (RV) function was evaluated by MPI. No subject had signs or symptoms of clinically overt heart failure. Systolic function was normal in all subjects. Among the echocardiographic indices of LV diastolic function the peak E velocity, E velocity/A velocity ratio, isovolumetric relaxation time, MPI and TFPV in the RA group were significantly different from those of the controls ( P < 0.05). However, we did not observe a significant difference in RV echocardiographic indices between the two groups. Our results show that there is LV diastolic dysfunction in patients with long-standing RA. The lack of a history of cardiotoxic antirheumatic drug use among our patients suggests that this abnormality is due to RA itself. PMID- 12740667 TI - A case of equine granulocytic ehrlichiosis provides molecular evidence for the presence of pathogenic anaplasma phagocytophilum (HGE agent) in Germany. AB - Based on seroprevalence studies and tick infection rates, tick-borne human granulocytic ehrlichiosis (HGE) is thought to occur in Germany, but to date no clinical case has been detected. Reported here are the first ehrlichial sequences derived from a German horse that fell ill with granulocytic ehrlichiosis. The analysis of three different genes (16S rRNA gene, groESL, and ankA) revealed up to 100% identity with ehrlichial sequences derived from patients with HGE in other countries or from infected ticks in Germany. Thus, the current lack of clinical cases of HGE in Germany is unlikely to result from the absence of pathogenic granulocytic ehrlichiae strains in German ticks. PMID- 12740670 TI - Fever of unknown origin: a review of 20 patients with adult-onset Still's disease. AB - In this study we aimed to investigate the findings in patients with adult-onset Still's disease (AOSD) admitted with fever of unknown origin (FUO) during the last 18 years in our unit, in order to discover the ratio of such patients to all patients with FUO during the same period, and to determine the clinical features of AOSD in FUO. The number and the aetiologies of the patients with FUO diagnosed between 1984 and 2001, and the clinical features of those with AOSD, were taken from the patient files. The diagnosis of AOSD was reanalysed according to the diagnostic criteria of Cush et al. [11]. The presumed diagnoses before a diagnosis of AOSD was established were also noted. The chi(2) and Fisher's exact tests were used for statistical analysis. We studied 130 patients with a diagnosis of FUO, 36 (28%) of whom had collagen vascular diseases. Of these 36 patients, 20 (56%, 12 female, 8 male, mean age 34 years, range 16-65) had AOSD. Clinical and laboratory findings were as follows: fever (100%), arthralgia (90%), rash (85%), sore throat (75%), arthritis (65%), myalgia (60%), splenomegaly (40%), hepatomegaly (25%), lymphadenopathy (15%), anaemia (65%), neutrophilic leukocytosis (90%), increased erythrocyte sedimentation rate (100%), elevated transaminase levels (65%), a negative RF (100%), and a negative FANA (80%). Antibiotics had been prescribed in 18 (90%) of cases. The presumed infectious diagnoses were streptococcal tonsillitis/pharyngitis (50%), infective endocarditis (four patients), sepsis (two patients) and acute bacterial meningitis (two patients). The presumed non-infectious diagnoses were acute rheumatic fever (three patients), seronegative rheumatoid arthritis (two patients) and polymyositis (two patients). Sixteen patients were followed for a mean duration of 30 months (range 2-59). A remission was obtained with indomethacin in three cases (19%), and with prednisolone in the remainder. Relapse was detected in three cases (19%). AOSD is one of the most frequent aetiologies of FUO. During the diagnostic course of a patient with FUO, a maculopapular rash and/or arthralgia and/or sore throat should raise the suspicion of AOSD. Because the disease has heterogeneous clinical findings, certain bacterial infections (e.g. streptococcal pharyngitis and sepsis) are generally considered and the prescribing of antibiotics is common. PMID- 12740671 TI - Cervical spine involvement in ankylosing spondylitis. AB - Our objective was to study cervical spine involvement in a Moroccan population of ankylosing spondylitis (AS) patients and evaluate correlations with disease symptomatic and structural severity. Patients were prospectively enrolled for a 1 year period. Clinical, biological, and radiological data were collected. The risk of cervical spine involvement was estimated using the Kaplan-Maier method. Sixty one patients were enrolled: 38 males (62.2%) and 23 females of mean (SD) age 35.1 years [11] (range 17-66). The mean disease duration was 10.6 years [7] (0.5-30). Forty-three patients (70.4%) had a history of neck pain. Radiological involvement was present in 33 cases (54%). The concordance between clinical and radiological involvement was statistically significant (kappa=0.49; P<10(-6)). The risk of cervical spine involvement with regard to disease duration showed that 19.6% of patients had radiological involvement after 5 years, 29.9% after 10 years, 45.1% after 15 years and 70.0% after 20 years. Comparison between patients with and without cervical spine radiological involvement showed no difference in age of onset or sex. There was statistical difference in symptomatic severity parameters (Schober, chest expansion, BASMI, BASFI, BASDI, BASG) and structural severity parameters (lumbar syndesmophytes score, BASRI). Our study confirms the greater severity of AS in North African countries. Cervical spine involvement increases with age and disease duration in AS and is more frequent in symptomatic and structural severe forms of the disease. PMID- 12740672 TI - Serum sIL-2r, IL-6, IL-10 and TNF-alpha level in familial Mediterranean fever patients. AB - In this study we investigated cytokine levels in patients with familial Mediterranean fever (FMF). Twenty patients and 20 healthy controls were included. Ten patients had acute attacks of FMF, whereas the other 10 were in the silent period. Patients with the acute exacerbation of FMF had higher soluble interleukin-2 receptor (sIL-2r), interleukin-6 (IL-6), tumour necrosis factor alpha, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) and fibrinogen levels than those in the silent period ( P<0.001) and controls ( P<0.001). In patients with acute attacks of FMF, interleukin-10 (IL-10) levels were not significantly different from those in the other patients or the controls ( P>0.05). In FMF patients IL-6, TNF-alpha, sIL-2r, ESR, CRP and fibrinogen levels increased with the acute-phase reaction, especially in the attack period. On the other hand, anti-inflammatory cytokine IL-10 levels did not increase as much as did the inflammatory cytokines. The balance between the cytokines may help us to understand the pathophysiology of FMF and to develop therapies. We conclude that the levels of the acute-phase reactants and the cytokines could be useful for diagnosis of acute exacerbations, follow-up and treatment. However, the cost of cytokine measurement analyses seems disadvantageous at present. PMID- 12740673 TI - Interleukin-13 in systemic sclerosis: relationship to nailfold capillaroscopy abnormalities. AB - The aim of this study was to investigate whether interleukin-13 (IL-13) serum levels correlate to different nailfold capillaroscopy (NC) findings in patients with systemic sclerosis (SSc). IL-13 serum levels were measured using an ELISA method. The following NC abnormalities were considered: the presence of giant loops, haemorrhages, loss of capillaries, disorganisation of the vascular array, ramified/bushy capillaries and sludging of blood. A semiquantitative rating scale was adopted to score these changes, as well as a rating system for avascular areas and three morphological NC patterns ('early', 'active' and 'late'). Mean capillary density was determined by counting the total number of capillaries in a 1 mm length, and the arterial and venous diameters of the capillary as well as the total loop diameter were measured. In SSc patients IL-13 serum levels were significantly higher than in controls ( P < 00.1), whereas in patients with ( n=8) and without ( n=24) abnormal IL-13 serum levels (>17 pg/ml) the comparison of the NC features showed significantly relevant differences concerning a more frequent 'active' NC pattern ( P < 0.02), the presence of haemorrhages ( P < 0.0037) and sludging of blood ( P < 0.038), as well as larger total loop ( P < 0.036) and arterial ( P < 0.03) diameters, in those patients with elevated IL-13 serum levels. The study confirmed that IL-13 serum levels are higher in the sera of patients with SSc, and shows for the first time the significant correlations between this serological finding and some of the main relevant SSc capillaroscopic features, leading us to believe that this cytokine not only seems to sustain the immunological and fibrotic process of SSc, but might have a role in determining the more severe microvascular lesions in this disease. PMID- 12740674 TI - Relationship of interleukin-12 and interleukin-13 imbalance with class-specific rheumatoid factors and anticardiolipin antibodies in systemic lupus erythematosus. AB - The aim of the study was to evaluate whether the imbalance between IL-12 and IL 13 serum levels, reflecting Th1/Th2 activity, is related to class-specific circulating rheumatoid factors (RF) and anticardiolipin (aCL) antibodies in SLE. Using ELISA we measured serum IL-12, IL-13, RF and aCL antibodies in 73 SLE patients and 20 healthy controls. The determination of IL-12/IL-13 ratio showed that IL-12 levels were above (group A), equal to (group B) or below (group C) IL 13 levels in 71.2%, 15.1% and 13.7% of SLE patients, respectively. IgM-RF levels were significantly higher in group C than in groups A ( P < 0.002) and B ( P < 0.019). Group C had also higher IgM-aCL levels than group A ( P < 0.04). No relationship between IL-12/IL-13 ratio and clinical or other laboratory parameters was found. It was concluded that the increased levels of both IgM-RF and IgM-aCL in patients with prevalent Th2 activity suggest that the predominance of Th2 over Th1 could drive autoantibody production in SLE patients. PMID- 12740675 TI - Efficacy of intra-articular sodium hyaluronate in the treatment of knee osteoarthritis. AB - To assess the efficacy of intra-articular hyaluronic acid in patients with knee osteoarthritis, sixty female patients with knee osteoarthritis were randomised to three weekly intra-articular injections of 30 mg sodium hyaluronate (Na HA) with a high molecular weight (1.0 to 2.9 million Da) or 40 mg 6-methylprednisolone acetate (6-MPA). The clinical assessments included pain at rest, at weight bearing and on walking, Lequesne Index and active range of knee flexion. Assessments were done at baseline, at week 4, and at months 3 and 6. A significant decrease in VAS scores for pain at rest, at weight-bearing and pain on walking, and in Lequesne index was found in both groups at week 4 when compared to baseline and there was no significant differences between the two groups. However, at 3(rd) month improvement in all pain scores and Lequesne index was found in favour of hyaluronic acid. At 6(th), no significant difference was found between the treatment groups. Improvement in pain was accompanied by an increase in joint flexion at week 4 and at month 3 in both groups. Both treatments were well-tolerated. The results showed that both intra-articular hyaluronic acid and 6-MPA treatments provide clinically significant improvement and demonstrated that Na HA has a long-term beneficial effect in patients with knee osteoarthritis. PMID- 12740677 TI - A new mathematical model based on clinical and laboratory variables for the diagnosis of Sjogren's syndrome. AB - Sjogren's syndrome (SS) is a systemic autoimmune disease that mainly affects exocrine glands. A diagnosis of SS in its early stages has a potential clinical relevance, but it is difficult and cannot be made solely on clinical grounds. Several sets of diagnostic criteria have been proposed, but none has met with a general consensus. Minor salivary gland has been judged to be the "gold standard" for the diagnosis of SS. However, it is a painful procedure and has a small but significant proportion of both false positive and false negative results. The aim of our study was to develop a simple mathematical score that uses clinical and laboratory variables for diagnosing SS, thereby reducing the need of minor salivary gland. The following variables were included in the model: ANA, SS-A/SS B, Schirmer's Test/BUT, C3/C4, serum gammaglobulin levels. One hundred consecutive individuals reporting clinical syndromes consistent with a sicca syndrome were included in the study. The application of our multifactorial mathematical model has shown a high predictive value for SS vs controls or vs patients with other autoimmune disorders (Sensitivity 93%, Specificity 100%), with an estimated minor salivary gland reduction of 77%. We conclude that our mathematical model can be considered a useful non-invasive approach for diagnosing Sjogren's Syndrome and recommend its validation on a larger scale. PMID- 12740676 TI - Comparison of two different viscosupplements in knee osteoarthritis -- a pilot study. AB - A pilot study was designed to investigate the efficacy of two different hyaluronic acid preparations combined to physical therapy in patients with knee osteoarthritis in terms of reduction in pain and disability and muscle strengthening. Thirty-seven patients with symptomatic osteoarthritis of the knees were randomly assigned into three groups. First group received a lower molecular weight hyaluronic acid plus physical therapy, second group received a higher molecular weight hyaluronic acid plus physical therapy, and the third group received physical therapy alone. The isokinetic knee muscle strengths and index of severity for osteoarthritis of the knee scores were evaluated at baseline, at the end of treatment (3 weeks) and at 3 months of follow up. At both short-term (3 weeks) and long-term (3 months) evaluations, index of severity for osteoarthritis of the knee scores were reduced in all three groups, while there was no significant muscle strengthening. PMID- 12740678 TI - A double-blind, multicentre, randomised clinical trial comparing the efficacy and tolerability of aceclofenac with diclofenac resinate in patients with acute low back pain. AB - The efficacy and tolerability of aceclofenac was compared with diclofenac resinate in a double-blind, multicentre randomised study in patients with acute low back pain suffering from degenerative spinal disorders. The study included 227 patients randomised to receive either aceclofenac 2 x 100 mg daily or diclofenac resinate 2 x 75 mg daily for up to 10 days. The primary objective was to demonstrate the clinical non-inferiority of the analgesic efficacy of aceclofenac compared with diclofenac resinate, as assessed by changes from baseline in the visual analogue scale (0-100 mm) pain score, at rest and at visit 3 (final visit on day's 8-10). Secondary objectives included the time to early cure (resolution of pain) and global assessment of tolerability. Mean change in pain score at rest, and as visit 3, compared with baseline, was 61.6 mm (SD 24.5) for the aceclofenac group ( n = 100) and 57.3 mm (SD 22.8) for the diclofenac resinate group ( n = 105) in the per-protocol population. Similar changes were observed in the intention-to-treat population. Between-group differences of 4.5 mm and 5.5 mm for the per-protocol and intention-to-treat populations, respectively, demonstrated clinical non-inferiority of aceclofenac compared with diclofenac resinate. Furthermore, there was evidence for superiority of aceclofenac over diclofenac resinate in terms of statistical significance, as the one-sided 97.5% confidence interval was above -10 mm and 0 mm. In the intention to treat population, a total of six aceclofenac-treated patients discontinued their medication owing to early cure, compared with only one patient receiving diclofenac resinate. Seventeen aceclofenac- (14.9%), and 18 diclofenac resinate treated patients (15.9%) reported at least one adverse event. However, the total number of adverse events reported was lower in patients receiving aceclofenac (22 versus 31 in the diclofenac resinate group). In conclusion, non-inferiority of the analgesic efficacy of aceclofenac compared with diclofenac resinate was demonstrated in patients with localised, uncomplicated acute lumbosacral pain. For the reduction in pain levels from baseline there was also evidence for superiority of aceclofenac compared with diclofenac resinate in terms of statistical significance, although this difference was not considered clinically relevant. The results also showed a trend towards a better safety and tolerability profile of aceclofenac over diclofenac resinate from a clinical point of view. PMID- 12740679 TI - Mitral valve vegetation and cerebral emboli in a primary antiphospholipid syndrome patient who had hepatitis C virus infection: report of a case and review of the literature. AB - We report the case of 36-year-old woman who came to us with a history of recurrent miscarriages and who was later diagnosed as having primary antiphospholipid syndrome (PAPS) and chronic hepatitis C virus (HCV) infection. The patient was referred to us with generalised seizures; cranial MRI revealed multiple embolic infarcts in both frontal lobes and a focal cortical infarct in the left frontoparietal lobe. Her echocardiography showed mitral valve vegetation and insufficiency. The patient was put on oral anticoagulant therapy and during her 8-month follow-up period no thrombotic events occurred. We report this case because it was the first in which PAPS, valvular disease, a cerebral embolic event and HCV infection were coexistent in the same patient. We also review other cases in which there was valvular vegetation and a cerebral ischaemic event associated with PAPS. PMID- 12740680 TI - Successful treatment of mesenteric vasculitis caused by Henoch-Schonlein purpura with methylprednisolone pulse therapy. AB - Although mesenteric vasculitis due to Henoch-Schonlein purpura (HSP) is relatively uncommon, it is the most life-threatening manifestation associated with high mortality. We describe a 15-year-old boy with HSP who had massive gastrointestinal bleeding and ileus but delayed onset of the purpuric rash. Abdominal ultrasonography revealed thickening of both small and large intestinal walls, and CT found prominent mesenteric vessels with comb sign and double wall of the bowel. These findings were consistent with mesenteric vasculitis and bowel ischaemia. The ischaemic intestine recovered after methylprednisolone pulse therapy and surgical intervention was avoided. Our report suggests that corticosteroid pulse therapy may help controlling HSP with massive gastrointestinal haemorrhage and ischaemic bowel due to widespread mesenteric vasculitis. PMID- 12740681 TI - Localised gastrocnemius myositis in Crohn's disease. AB - A19-year-old woman presented with pain and tenderness in both calves associated with pyrexia and neutrophil leukocytosis. Gastrocnemius muscle biopsy showed a non-specific lymphocytic myositis and she was found to have positive c-ANCA, in the absence of other evidence of systemic vasculitis. Subsequent investigation of her gastrointestinal tract revealed extensive Crohn's disease. The myositis responded promptly to treatment with prednisolone 0.5 mg/kg. A review of the literature showed that localised calf pain in the setting of Crohn's disease can be caused by non-specific myositis, granulomatous myositis or vasculitis. It is proposed that the "gastrocnemius myalgia syndrome" be included in the typical - albeit rare - extraintestinal manifestations that may herald the appearance of inflammatory bowel disease. PMID- 12740682 TI - Gallium-67 scintigraphy in the diagnosis and management of chronic sarcoid myopathy. AB - The authors describe a patient with chronic sarcoid myopathy. Except for the presence of left posterior synechia, no other organ involvement was observed. Gallium-67 ((67)Ga) scintigraphy showed many intense nodular uptake areas in both the upper and lower extremities. Treatment with oral prednisolone 30 mg/day resulted in a marked improvement on (67)Ga scintigraphy. This case suggests that (67)Ga scintigraphy is useful for the differential diagnosis of systemic myopathies and also for monitoring the effect of glucocorticoid treatment. PMID- 12740684 TI - Rheumatoid nodules indicating seronegative rheumatoid arthritis in a patient with gout. AB - The association of gout and rheumatoid arthritis is rare. We report the case of a patient with gout who presented with rheumatoid nodules indicating seronegative rheumatoid arthritis. PMID- 12740683 TI - Polyarteritis nodosa limited to calf muscles: a case report and review of the literature. AB - We describe an unusual case of a 38-year-old woman with a localised form of polyarteritis nodosa (PAN) manifested by acute onset of severe calf pain. Magnetic resonance imaging of the lower legs showed abnormal signal intensity of the outer calves muscle. Biopsies of the gastrocnemius revealed an acute necrotising arteritis with marked non-specific fasciitis. The diseased muscle improved with corticosteroid administration alone. The fascial involvement likely contributed to the severity of the clinical features of PAN, which may be related to a previous hepatitis B virus infection. PMID- 12740685 TI - Cogan's syndrome presenting as Sjogren's syndrome followed by acute aortic regurgitation. PMID- 12740686 TI - Treatment of adult-onset Still's disease with leflunomide and chloroquine combination in two patients. PMID- 12740687 TI - Critical hypokalemic renal tubular acidosis due to Sjogren's syndrome: association with the purported immune stimulant echinacea. PMID- 12740688 TI - A case of primary antiphospholipid syndrome who developed acute myocardial infarction followed by early-onset pre-eclampsia. PMID- 12740689 TI - A case of juvenile systemic sclerosis with disease onset at six months old. AB - Systemic sclerosis (SSc) is a generalised connective tissue disorder characterised by sclerotic changes in the skin and internal organs. The occurrence of SSc in childhood is rare, with childhood-onset disease representing 3% of all SSc cases, and fewer than 100 such patients have been reported in the world literature to date [1]. We report a case of juvenile SSc with disease onset at the age of 6 months. To our knowledge, this case is the youngest in the world literature. PMID- 12740690 TI - A technique for minor salivary gland biopsy used in the diagnosis of Sjogren's syndrome. PMID- 12740691 TI - Association of single-nucleotide polymorphisms in the polymeric immunoglobulin receptor gene with immunoglobulin A nephropathy (IgAN) in Japanese patients. AB - Immunoglobulin A nephropathy (IgAN) is a primary glomerulonephritis of common incidence world-wide whose etiology and pathogenesis remain unresolved, although genetic factors are assumed to be involved in the development and progression of this disease. To identify genetic variations that might confer susceptibility to IgAN, we performed a case-control association study involving 389 Japanese IgAN patients and 465 controls. Genome-wide analysis of approximately 80,000 single nucleotide polymorphisms (SNPs) identified a significant association between IgAN and six SNPs located in the PIGR (polymeric immuoglobulin receptor) gene at chromosome 1q31-q41. One of them, PIGR-17, caused an amino-acid substitution from alanine to valine at codon 580 (chi(2)=13.05, P=0.0003, odds ratio [OR] =1.59, 95% confidence interval [95% CI] =1.24-2.05); the OR of minor homozygotes to others was 2.71 (95% CI=1.31-5.61). Another SNP, PIGR-2, could affect promoter activity (chi(2)=11.95, P=0.00055, OR=1.60, 95% CI=1.22-2.08); the OR of minor homozygotes to others was 2.08 (95% CI=0.94-4.60). Pairwise analyses demonstrated that all six SNPs were in almost complete linkage disequilibrium. Biopsy specimens from IgAN patients were positively stained by antibody against the secretory component of PIGR, but corresponding tissues from non-IgAN patients were not. Our results suggest that a gene associated with susceptibility to IgAN lies within or close to the PIGR gene locus on chromosome 1q in the Japanese population. PMID- 12740692 TI - Repair of intercostal pulmonary herniation. AB - Intercostal pulmonary herniation occurring years after blunt thoracic trauma is a rare phenomenon. We report on the case of a 66-year-old patient who developed a pulmonary herniation 2 years after a seat-belt injury. Thoracotomy was performed, and the thoracic wall defect was closed with approximating periostal absorbable sutures. The postoperative course was uneventful. Different surgical approaches and the use of prosthetic patches are discussed. Periostal fixation of the adjacent ribs with absorbable sutures is usually sufficient for herniation repair. In cases in which prosthetic meshes are needed, the application of PTFE might produce the best results with the least complications. PMID- 12740693 TI - Composite resin fillings and inlays. An 11-year evaluation. AB - The purpose of this randomized, clinical study was to evaluate the clinical performance of composite resin materials used for fillings and indirect inlays. Twenty-eight sets of five class II restorations (two fillings, three inlays) were placed in 88 premolars and 52 molars in 28 adults. Brilliant Dentin and Estilux Posterior were used for both fillings and inlays, and SR-Isosit for inlays only. After 11 years, 27 sets of restorations (96%) were evaluated clinically using modified United States Public Health Service criteria. Replaced or repaired restorations were observed in 16% of the fillings and 17% of the inlays, and a further 5% of the restorations were replaced for reasons not related to the restoration. The remaining 107 restorations exhibited optimal ratings in 30% of the fillings and 12% of the inlays (P<0.05) and acceptable ratings in 70% and 88%, respectively. The reasons for failure were fracture of restoration (four fillings, five inlays), secondary caries (two fillings, four inlays), fracture of tooth (two inlays), loss of proximal contact (two fillings), and loss of restoration (one inlay). Failures were seen more frequently in molar than premolar restorations (P<0.05), with no significant difference between fillings and inlays or between the five types of restoration (P>0.05). PMID- 12740694 TI - Freeze/thaw stress in Ceanothus of southern California chaparral. AB - Freeze/thaw stress was examined in chaparral shrubs of the genus Ceanothus to determine the interactive effects of freezing and drought and to consider which is the more vulnerable component, the living leaves (symplast) or the non-living water transport system (apoplast). We hypothesized that where Ceanothus species co-occurred, the more inland species C. crassifolius would be more tolerant of low temperatures than the coastal species C. spinosus, both in terms of leaf survival (LT(50), or the temperature at which there is 50% loss of function or viability) and in terms of resistance to freezing-induced embolism (measurements of percent loss hydraulic conductivity due to embolism following freeze/thaw). Cooling experiments on 2 m long winter-acclimated shoots resulted in LT(50) values of about -10 degrees C for C. spinosus versus -18 degrees C for C. crassifolius. Freeze-thaw cycles resulted in no change in embolism when the plants were well hydrated (-0.7 to -2.0 MPa). However, when plants were dehydrated to -5.0 MPa, C. spinosus became 96% embolized with freeze/thaw, versus only 61% embolism for C. crassifolius. Stems of C. crassifolius became 90% and 97% embolized at -6.6 and -8.0 MPa, respectively, meaning that even in this species, stems could be more vulnerable than leaves under conditions of extreme water stress combined with freeze/thaw events. The dominance of C. crassifolius at colder sites and the restriction of C. spinosus to warmer sites are consistent with both the relative tolerance of their symplasts to low temperatures and the relative tolerance of their apoplasts to freeze events in combination with drought stress. PMID- 12740695 TI - A case of Mycoplasma hominis meningo-encephalitis in a full-term infant: rapid recovery after start of treatment with ciprofloxacin. AB - The role of Mycoplasma hominisas a causative agent for neonatal sepsis and meningitis is still unclear. Meningitis secondary to M. hominisis well-described in the literature; however, M. hominiscan also be isolated from cerebrospinal fluid (CSF) obtained from infants without signs of meningitis. We present a case of a full-term infant with meningo-encephalitis with seizures, epileptic activity on the EEG, inflammation of brain tissue on a CT scan, and cloudy CSF containing elevated cell counts, decreased glucose levels and elevated protein levels. M. hominiswas identified from the CSF by culture and by polymerase chain reaction (PCR) as the only possible causative agent. Furthermore, while empiric antibiotic and antiviral treatment for neonatal sepsis had failed, the meningo-encephalitis promptly responded upon antibiotic treatment with ciprofloxacin (20 mg/kg per day i.v.), to which M. hominisis susceptible. CONCLUSION: A meningo-encephalitis developed due to infection with M. hominisin a full-term infant, from which he recovered rapidly after start of treatment with ciprofloxacin. PMID- 12740696 TI - Concomitant anuric post-streptococcal glomerulonephritis and autoimmune hemolytic anemia. PMID- 12740697 TI - A genomewide survey of developmentally relevant genes in Ciona intestinalis. X. Genes for cell junctions and extracellular matrix. AB - Cell junctions and the extracellular matrix (ECM) are crucial components in intercellular communication. These systems are thought to have become highly diversified during the course of vertebrate evolution. In the present study, we have examined whether the ancestral chordate already had such vertebrate systems for intercellular communication, for which we have searched the genome of the ascidian Ciona intestinalis. From this molecular perspective, the Ciona genome contains genes that encode protein components of tight junctions, hemidesmosomes and connexin-based gap junctions, as well as of adherens junctions and focal adhesions, but it does not have those for desmosomes. The latter omission is curious, and the ascidian type-I cadherins may represent an ancestral form of the vertebrate type-I cadherins and desmosomal cadherins, while Ci-Plakin may represent an ancestral protein of the vertebrate desmoplakins and plectins. If this is the case, then ascidians may have retained ancestral desmosome-like structures, as suggested by previous electron-microscopic observations. In addition, ECM genes that have been regarded as vertebrate-specific were also found in the Ciona genome. These results suggest that the last common ancestor shared by ascidians and vertebrates, the ancestor of the entire chordate clade, had essentially the same systems of cell junctions as those in extant vertebrates. However, the number of such genes for each family in the Ciona genome is far smaller than that in vertebrate genomes. In vertebrates these ancestral cell junctions appear to have evolved into more diverse, and possibly more complex, forms, compared with those in their urochordate siblings. PMID- 12740698 TI - A genomewide survey of developmentally relevant genes in Ciona intestinalis. IX. Genes for muscle structural proteins. AB - Ascidians are simple chordates that are related to, and may resemble, vertebrate ancestors. Comparison of ascidian and vertebrate genomes is expected to provide insight into the molecular genetic basis of chordate/vertebrate evolution. We annotated muscle structural (contractile protein) genes in the completely determined genome sequence of the ascidian Ciona intestinalis, and examined gene expression patterns through extensive EST analysis. Ascidian muscle protein isoform families are generally of similar, or lesser, complexity in comparison with the corresponding vertebrate isoform families, and are based on gene duplication histories and alternative splicing mechanisms that are largely or entirely distinct from those responsible for generating the vertebrate isoforms. Although each of the three ascidian muscle types - larval tail muscle, adult body wall muscle and heart - expresses a distinct profile of contractile protein isoforms, none of these isoforms are strictly orthologous to the smooth-muscle specific, fast or slow skeletal muscle-specific, or heart-specific isoforms of vertebrates. Many isoform families showed larval-versus-adult differential expression and in several cases numerous very similar genes were expressed specifically in larval muscle. This may reflect different functional requirements of the locomotor larval muscle as opposed to the non-locomotor muscles of the sessile adult, and/or the biosynthetic demands of extremely rapid larval development. PMID- 12740699 TI - A genomewide survey of developmentally relevant genes in Ciona intestinalis. VII. Molecules involved in the regulation of cell polarity and actin dynamics. AB - In the present study, genes involved in the pathways that establish cell polarity and cascades regulating actin dynamics were identified in the completely sequenced genome of Ciona intestinalis, a basal chordate. It was revealed that the Ciona genome contains orthologous genes of each component of aPKC-Par and PCP pathways and WASP/WAVE/SCAR and ADF/cofilin cascades, with less redundancy than the vertebrate genomes, suggesting that the conserved pathways/cascades function in Ciona development. In addition, the present study found that the orthologous proteins of five gene groups (Tc10, WRCH, RhoD, PLC-L, and PSKH) are conserved in humans and Ciona but not in Drosophila melanogaster, suggesting a similarity in the gene composition of Ciona to that of vertebrates. Ciona intestinalis, therefore, may provide refined clues for the study of vertebrate development and evolution. PMID- 12740701 TI - Isolated dorsal dislocation of the intermediate cunieform: a case report and review of the literature. AB - We present one very rare case of patient with an isolated dorsal dislocation of the middle cuneiform (closed injury) sustained after a fall. He was treated surgically through a dorsal approach and fixation with K-wires after manual reduction was unsuccessful. A cast was applied for a total of 6 weeks followed by active range-of-motion exercises. At the 18-month follow-up, he was asymptomatic. PMID- 12740702 TI - Which one is at risk in intraoperative fluoroscopy? Assistant surgeon or orthopaedic surgeon? AB - BACKGROUND: Orthopaedic surgeons use intraoperative portable fluoroscopy and roentgenography. The present study was an attempt to find out if there is a difference between the occupational radiation exposure to the orthopaedic surgeon and assistant surgeon in the operating room while using intraoperative fluoroscopy or radiographic control and to measure it. METHODS: During a 3-month period, 107 consecutive operations were monitored for radiation exposure. At monthly intervals, the radiation doses were measured in millirem and recorded. The distance of the orthopaedic surgeon and the assistant surgeon from the X-ray source were noted in every fluoroscopic check. The orthopaedic surgeon was always at a safe distance (more than 90 cm), but the assistant surgeon always stood nearby (10 cm) the X-ray source for positioning of the patient. RESULTS: The radiation exposure according to the badge on the shoulder was consecutively 3, 4, 3 mrem for the orthopaedic surgeon and 20, 19, 22 mrem for the assistant surgeon. The radiation exposure according to the badges on the anaesthetic machine, in the room and under the apron of the orthopaedic surgeon were all zero, whereas the readings of the badge under the apron of the assistant surgeon were 7, 6, 5 mrem consecutively. CONCLUSION: Our findings show that although the radiation exposure during orthopaedic operations is below the recommendations of the European Committee on Radiation Protection, there is a higher risk of exposure for the assistant surgeon. It has to be kept in mind that there could be morphological and functional damage in cells exposed to radiation. Therefore, we should continue to use appropriate shielding precautions in view of the unknown long term risks. PMID- 12740700 TI - Effects of caffeine on potassium currents in isolated rat ventricular myocytes. AB - Rapid exposure of cardiac muscle to high concentrations of caffeine releases Ca(2+) from the sarcoplasmic reticulum (SR). This Ca(2+) is then extruded from the cell by the Na(+)/Ca(2+) exchanger. Measurement of the current carried by the exchanger (I(Na/Ca)) can therefore be used to estimate of the Ca(2+) content of the SR. Previous studies have shown that caffeine, however, can also inhibit K(+) currents. We therefore investigated whether the inhibitory effects of caffeine on these currents could contaminate measurements of I(Na/Ca). Caffeine caused partial inhibition of the inward rectifier K(+) current (I(K1)): the outward current at -40 mV was 1.15+/-0.24 pA/pF in control and decreased to 0.34+/-0.15 pA/pF in the presence of 10 mmol/l caffeine (P<0.05, n=15). This was similar to the effect of caffeine on the holding current observed at -40 mV in the absence of K(+) channel block and could therefore account for the contaminating effects of caffeine observed during measurements of I(Na/Ca). Moreover, caffeine also partially inhibited the transient outward ( I(to)) and the delayed rectifier (I(K)) K(+) currents. PMID- 12740703 TI - A pulmonary allergic reaction after injection sclerotherapy for hemorrhoids. PMID- 12740704 TI - Experimental study of the embryogenesis of gastrointestinal duplication and enteric cyst. AB - The theory of gastrointestinal duplication and enteric cyst embryogenesis was verified by examining the developmental process of this experimentally induced anomaly. In Cynopus pyrrhogaster (amphibian) embryos (stage 18), the dorsal midline structures (including the neural plate and notochord) were split regionally to induce partial separation of the notochord and gut anlage endoderm herniation between the split elements of the notochord. Following this procedure, the embryonic development was traced morphologically and histologically. Control embryos were cultured without the procedure. Following the incubation and breeding period, gastrointestinal duplication and enteric cysts were observed with vertebral anomaly, spina bifida, split cord malformation and subcutaneous manifestations in the mature animals. The combination of anomalies that was observed in these experimental animals is consistent with that found in "split notochord syndrome." No abnormal morphology or histology was observed in the control group. The embryogenetic theory of gastrointestinal duplication and enteric cysts was thus verified by simulating the partial separation of the notochord, which induced split notochord syndrome in laboratory animals. The results indicate that gastrointestinal duplication and enteric cysts may arise through a process of herniation of the gut anlage endoderm between split elements of the notochord. PMID- 12740705 TI - Urinary ascites in infancy: varied etiologies. AB - Three cases of urinary ascites are presented, each with a different underlying aetiology. The age and modes of presentation also varied and management strategies were accordingly tailored to each patient's clinical requirements. All 3 patients survived and subsequently were discharged with good renal function. Although a rare condition, infants with urinary ascites can present as clinical emergencies in need of prompt resuscitation with subsequent drainage of the urine and decompression of the urinary tract. The ultimate management regime will vary and depend upon site of urinary extravasation and underlying aetiology. PMID- 12740706 TI - Sentinel lymph node mapping and biopsy: a potentially valuable tool in the management of childhood extremity rhabdomyosarcoma. AB - Multimodal therapy, involving surgery, chemotherapy and radiation, now dominates the management of rhabdomyosarcoma (RMS) in childhood. Yet, despite improvements in these practices, extremity tumors continue to fare relatively poorly. Several investigators have identified prognostic factors that can be used to direct therapy and predict outcome. These factors include histology and metastatic disease, the latter requiring accurate staging to identify. The presence of lymph node metastases has been shown to be of prognostic significance and is incorporated into pre-treatment staging schemes. Up to 50% of all surgically evaluated nodes and 17% of clinically negative nodes in extremity RMS may harbor tumor, underscoring the increased risk of understaging the disease if accurate lymph node dissection is not undertaken. Despite its importance, there appears to be no standard format by which regional nodal status is evaluated in extremity RMS. Sentinel lymph node mapping and biopsy are a minimally invasive technique, currently used in the staging of adult breast cancer and melanoma. In adults, the technique is associated with optimum nodal yield and low morbidity. We describe a case in which sentinel node mapping and dissection were used to easily and accurately stage a distal upper extremity alveolar RMS in a child with clinically and radiologically negative regional lymph nodes. The procedure yielded no positive nodes, was associated with no morbidity and spared the child more extensive radiotherapy. We propose the further evaluation of this simple and innovative technique in the overall management of this childhood malignancy. PMID- 12740707 TI - Intraventricular migration of a Rickham reservoir: endoscopic retrieval. AB - CASE REPORT: An unusual case of the intraventricular migration of a Rickham reservoir in a child and its subsequent removal by endoscopy is described. A newborn boy presenting with hydrocephalus secondary to intraventricular haemorrhage, had a Rickham reservoir inserted through a frontal burr hole, made adjacent to the anterior fontanel. Later, serial CT scans of the brain over a 3 year period revealed progressive migration of the Rickham reservoir into the lateral ventricle and then into the third ventricle. As the ventricles were also enlarged, they were explored endoscopically and the Rickham reservoir was removed. DISCUSSION: Intracranial migration of a Rickham reservoir is rare. It is suggested that the placement of a small burr hole, away from the fontanels and growing sutures and the use of anchoring sutures to hold the reservoir to the periosteum are important steps in avoiding this unusual complication. PMID- 12740708 TI - A critical analysis of 'normal' radionucleotide shuntograms in patients subsequently requiring surgery. AB - BACKGROUND: Shuntograms are performed when patients present with symptoms suggestive of, but inconclusive for, shunt malfunction, without confirmatory radiological evidence. METHODS: Shuntograms over the past 3.5 years were reviewed. Patient records were reviewed for revision in proximity to a negative (normal) study. RESULTS: One hundred and fifteen out of 149 tests were negative. Thirty-four surgeries (in 31 patients) occurred subsequent to a negative shuntogram. In 18 out of 34 revisions the shunt was functional: 13 surgeries were for overdrainage, 4 were for unrelated reasons with shunt function confirmed incidentally and 1 was an exploration for cognitive deterioration. In 16 cases (13 patients) the shunt was not functional: 12 had proximal catheter occlusion in which, on subsequent review, there was no ventricular reflux present and the remaining had distal malfunctions. CONCLUSIONS: The false negative rate for shuntograms was 16 out of 115 (14%) with proximal occlusion most common. This estimate of the predictive value of a normal flow study may influence the decision to revise a shunt. PMID- 12740709 TI - Multi-slice spiral CT in routine diagnosis of suspected acute left-sided colonic diverticulitis: a prospective study of 120 patients. AB - This prospective study evaluated the use of multi-slice CT (MSCT) for detection of clinically suspected left-sided colonic diverticulitis with regard to diagnosis, complications and alternative diagnoses. One hundred twenty patients with clinically suspected acute left-colonic diverticulitis underwent MSCT of the lower abdomen with IV contrast after rectal application of iodic contrast. The MSCT results were compared with histopathological and intraoperative findings or other radiological or endoscopic methods and clinical outcome. Acute diverticulitis was proven in 67 of the 120 (55.8%) patients, which was detected by MSCT with an accuracy of 98% (sensitivity 97%, specificity 98%). Contained perforation or abscess formation were detected with an accuracy of 96% (sensitivity 100%, specificity 91%) and 98% (sensitivity 100%, specificity 97%), respectively. In 31 of 120 (25.8%) patients diagnoses other than diverticulitis caused abdominal pain, which was correctly diagnosed by MSCT in 71%. The MSCT as well as other concurrently performed diagnostic methods showed normal findings and no causes for the patients symptoms in 22 of the 120 (18.4%) patients. Multi slice CT is reliable in detecting diverticulitis, including extracolic complications, and often reveals other diagnoses; therefore, MSCT is recommended as standard diagnostic procedure in suspected acute diverticulitis. PMID- 12740710 TI - Retrospective measurement of different size parameters of non-radiated rectal cancer on MR images and pathology slides and their comparison. AB - There are no non-invasive methods to assess the real tumor size in rectal cancer prior to surgery, especially following radio/chemotherapy. Magnetic resonance imaging is gaining increasing acceptance as the primary modality at many centers for evaluation of pelvic malignancies including rectal cancers. The aim of this study was to evaluate if the tumor size as assessed by stereological or metric means on MRI correlates to the corresponding pathologic findings. To our knowledge, no such previous work has been reported in the literature. From the Cancer Register Center, 18 patients in the age range of 39-90 years with rectal cancer who had complete preoperative MR with subsequent giant section pathological examinations of the resected bowel were included. The tumor size was measured on MR and histopathologic specimen using both a stereologic and a metric mode. The measured parameters included the maximum transverse area occupied by the tumor, thickness, width, and the length of tumor and the volume of the tumor measured in two different fashions by the product of area and length (al) or the product of thickness, width, and length (twl). The depth of tumor infiltration (T) and presence of local lymph node metastases (N) were also separately evaluated on the histopathologic specimen. There were 1, 4, 12, and 1 patients with tumor stages T1, T2, T3, and T4, respectively. The mean thickness, width, length, area, and volumes, al and twl, were 1.62, 2.8, and 4.78 cm, and 4.72 cm2, 26.29 cm3, and 20.07 cm3, respectively. Regression curves were drawn for above mentioned parameters. They showed some correlation with square correlation coefficient measuring between 0.38 and 0.82. The best correlation was seen for area (0.75) and volume measured by the product of area and length of the tumor (0.82). With the formula proposed from this material, we assume that rectal tumors can be measured on MR images using a metric model, especially area and the volume (the product of area and length), and then extrapolated to what we would expect from pathology, hence providing us with a tool where we could measure tumor response after neoadjuvant therapy. PMID- 12740712 TI - Interstitial involvement in children: "shaggy heart". PMID- 12740713 TI - A semi-quantitative RT-PCR method to readily compare expression levels within Botrytis cinerea multigenic families in vitro and in planta. AB - A straightforward and easy-to-apply semi-quantitative RT-PCR method was developed to study multigenic expression in the phytopathogenic fungus Botrytis cinerea. This procedure is based on the one-step reverse transcription-amplification of a specific transcript within total RNA and product amount determination by densitometric analysis of ethidium bromide fluorescence upon gel electrophoresis. The semi-quantitative analysis is achieved, at a fixed PCR cycle-number, within a range of total RNA concentrations that stays in the exponential phase of the PCR. Co-amplification of the transcript of interest with internal controls allowed comparison between different RNA samples. Using this method, we could demonstrate a differential regulation of chitin synthase genes during fungal growth and an effect of the culture carbon source on the expression of two pectin methylesterase genes in B. cinerea. Finally, the method was shown to be applicable to plant-infected tissue, making it a useful tool to detect pathogenicity genes in B. cinerea. PMID- 12740722 TI - Utilisation of structurally diverse organophosphonates by Streptomycetes. AB - A group of streptomycete strains was found able to utilise a wide range of structurally diverse phosphonates as a sole phosphorus source. No relation could be observed between ability to synthesise compounds containing a direct carbon-to phosphorus (C-P) bond and biodegradative potential towards phosphonates in the strains studied. Streptomyces morookaensis DSM 40565 could degrade 2-amino-4 phosphonobutyrate as a sole nitrogen and phosphorus source in a stereoselective like manner. This result suggests the existence of a new metabolic pathway for C P bond breakage. PMID- 12740721 TI - Treatment of cultured glioma cells with the EGFR-TKI gefitinib ("Iressa", ZD1839) increases the uptake of astatinated EGF despite the absence of gefitinib-mediated growth inhibition. AB - The EGFR-TKI (epidermal growth factor receptor tyrosine kinase inhibitor) gefitinib ("Iressa", ZD1839), a reversible growth inhibitor of EGFR-expressing tumour cells, has been shown to enhance the antitumour effect of ionising radiation, and also to increase the uptake of radioiodinated EGF. Thus, combination of gefitinib treatment and radionuclide targeting is an interesting option for therapy of brain tumours that are difficult to treat with conventional methods. The aim of this study was to evaluate how pre-treatment with gefitinib affects binding of astatinated EGF ((211)At-EGF) to cultured glioma U343 cells, which express high levels of EGFR. The growth of U343 cells in the presence of gefitinib was investigated, and it was found that gefitinib does not significantly inhibit the growth of these cells. Nevertheless, the uptake of (211)At-EGF in U343 cells was markedly increased (up to 3.5 times) in cells pre treated with gefitinib (1 microM). This indicates that a combination of gefitinib treatment and radionuclide targeting to EGFR might be a useful therapeutic modality, even for patients who do not respond to treatment with gefitinib alone. PMID- 12740723 TI - An alternative method to the osmotic stressing polymers: the osmomanometer. AB - Although stressing polymers have been widely and successfully used to determine the osmotic properties of solutes in aqueous media, the osmotic stress method presents some limitations. To overcome these drawbacks, an alternative and more direct method, which has been named the osmomanometer, is described in this letter. The osmotic pressure accessible by this method ranges typically from 1 to 30 kPa using a simple hydrostatic effect and can be extended to higher pressures by using pressurized gas. This method needs neither a pressure sensor nor calibration. PMID- 12740724 TI - Ataxia-telangiectasia: the pattern of cerebellar atrophy on MRI. AB - We describe MRI of the brain in 19 patients with ataxia-telangiectasia (AT) and correlate the appearances with the degree of neurologic deficit. We examined 10 male and nine female patients; 17 were aged between 2 and 12 years (mean 8 years) but a woman and her brother were 35 and 38 years old, and had a variant of AT. Ataxia was the first recognized sign of the disease in every patient. We detected the following patterns of cerebellar atrophy: in the youngest patient, aged 2 years, the study was normal; in the five next youngest patients 3-7 years of age, the lateral cerebellum and superior vermis showed the earliest changes of atrophy; and all but one of the other patients had moderate to marked diffuse atrophy of vermis and cerebellar hemispheres. There were 12 patients aged 9 years and above; one, who was normal, was 9 years old. The five patients who at the time of examination were unable to walk all had diffuse atrophy involving both vermis and cerebellar hemispheres. PMID- 12740714 TI - Regulation of phosphate acquisition in Saccharomyces cerevisiae. AB - Membrane transport systems active in cellular inorganic phosphate (P(i)) acquisition play a key role in maintaining cellular P(i) homeostasis, independent of whether the cell is a unicellular microorganism or is contained in the tissue of a higher eukaryotic organism. Since unicellular eukaryotes such as yeast interact directly with the nutritious environment, regulation of P(i) transport is maintained solely by transduction of nutrient signals across the plasma membrane. The individual yeast cell thus recognizes nutrients that can act as both signals and sustenance. The present review provides an overview of P(i) acquisition via the plasma membrane P(i) transporters of Saccharomyces cerevisiae and the regulation of internal P(i) stores under the prevailing P(i) status. PMID- 12740726 TI - Anticipatory postural adjustments while sitting: the effects of different leg supports. AB - The aim of this study was twofold, to analyze the effects of changes in body position and changes in the location of body supports on anticipatory postural adjustments (APAs). Eight healthy subjects were studied while sitting and standing. Subjects exerted upward or downward vertical force against an object attached to a rigid frame and released the object with a fast bilateral shoulder abduction movement. While sitting, four support conditions were studied: with and without feet support, and with anterior or posterior lower-leg supports. The electromyographic activity of leg and trunk muscles was recorded and quantified for APA activity. APAs in sitting with feet support were attenuated in the leg muscles (tibialis anterior, soleus, rectus femoris, and biceps femoris) but not in trunk muscles (erector spinae, rectus abdominis) when compared with standing. In the sitting task, series with and without feet support showed no difference in APAs. Anterior or posterior supports to the lower legs while sitting were associated with enhanced anticipatory activity in biceps femoris and rectus femoris muscles, respectively. However, trunk muscles showed similar anticipatory patterns across all the support conditions. We conclude that the central nervous system uses flexible, adaptive control strategies to adjust APAs to particular mechanical conditions induced by modification of a leg support. PMID- 12740725 TI - Development of postural adjustments during reaching in sitting children. AB - We evaluated the development of postural adjustments accompanying reaching movements in sitting children. Twenty-nine typically developing children aged, 2 11 years, and ten adults were studied with multiple surface electromyograms (EMGs) and kinematics during reaching in four conditions: sitting with the seat surface oriented horizontally with and without an additional task load, and sitting with the seat-surface tilted 15 degrees forward and 15 degrees backward. The development of postural adjustments during reaching in a sitting position turned out to have a non-linear and protracted course, which is not finished by the age of 11 years. The development of these adjustments is characterised by variation, yet specific developmental sequences could be distinguished. Firstly, the development of postural adjustments during reaching from the age of 2 years onwards lacked a preference for an en bloc strategy, which consists of an in concert activation of the direction-specific neck and trunk muscles. Secondly, anticipatory postural muscle activity, which was consistently present in adults, was virtually absent between 2 and 11 years of age. Thirdly, the data demonstrated that with increasing age the head gradually becomes the dominant frame of reference. In addition, the study suggested that, in terms of postural control, the forward-tilted position is the most efficient one. PMID- 12740727 TI - Vestibular, optokinetic, and cognitive contribution to the guidance of passive self-rotation toward instructed targets. AB - We ask how vestibular and optokinetic information is combined ("fused") when human subjects who are being passively rotated while viewing a stationary optokinetic pattern try to tell when they have reached a previously instructed angular displacement ("targeting task"). Inevitably such a task entices subjects to also draw on cognitive mechanisms such as past experience and contextual expectations. Specifically, because we used rotations of constant angular velocity, we suspected that they would resort, consciously or unconsciously, to extrapolation strategies even though they had no explicit knowledge of this fact. To study these issues, we presented the following six conditions to subjects standing on a rotatable platform inside an optokinetic drum: V, pure vestibular (passive rotation in darkness); O, pure optokinetic (observer motionless, drum rotating); VO, combined (passive rotation while viewing stationary drum); Oe, optokinetic extrapolation (similar to O, but drum visible only during first 90 degrees of rotation; thereafter subjects extrapolate the further course in their minds); VOe, combined extrapolation (similar to VO, but drum visible only during first 90 degrees ); AI, auditory imagination (rotation presented only metaphorically; observers imagine a drum rotation using the rising pitch of a tone as cue). In all conditions, angular velocities ( v(C)) of 15, 30, or 60 degrees /s were used (randomized presentation), and observers were to indicate when angular displacement (of the self in space or relative to the drum) had reached the instructed magnitude ("desired displacement", D(D); range 90-900 degrees ). Performance was analyzed in terms of the targeting gain ( G(T) = physical displacement at time of subjects' indication / D(D)) and variability (% E(R) = percentage absolute deviation from a subject's mean gain). In all six conditions, the global mean of G(T) (across v(C) and D(D)) was remarkably close to veracity, ranging from 0.95 (V) to 1.06 (O). A more detailed analysis of the gain revealed a trend of G(T) to be larger with fast than with slow rotations, reflecting an underestimation of fast and an overestimation of slow rotation. This effect varied significantly between conditions: it was smallest in VO, had intermediate values with the monomodal conditions V and O, and also with VOe, and was largest in Oe and AI. Variability was similar for all velocities, but depended significantly on the condition: it was smallest in VO, of intermediate magnitude in O, VOe, Oe, and largest in V and AI. Additional experiments with conditions V, O, and VO in which subjects repetitively indicated displacement increments of 90 degrees, up to a subjective displacement of 1080 degrees, yielded similar results and suggest, in addition, that the displacement perceptions measured at the beginning and during later phases of the rotation are correlated. With respect to the displacement perception during optokinetic stimulation, they also show that the gain and its variability are similar whether subjects feel stationary and see a rotating pattern, or feel rotated and see a stationary pattern (circular vection). We conclude that the vestibular and optokinetic information guiding the subjects' navigation toward an instructed target is not fused by straightforward averaging. Rather the subjects' internal velocity representation (which ultimately determines G(T)) appears to be a weighted average of (1) whatever sensory information is available and of (2) a cognitive default value reflecting the subjects' experiences and expectations. The less secure the sensory information (only one source as in V or O, additional degrading as in Oe or AI), the larger the weight of the default value. Vice versa, the better the information (e.g., two independent sources as in VO), the more the actual velocity and not the default value determines displacement perception. Moreover, we suggest that subjects intuitively proceeded from the notion of a constant velocity rotation, and therefore tended to carry on the perception built up during the beghe perception built up during the beginning of a rotation or, in the case of vestibular navigation, to compensate for the decaying vestibular cue by means of an internal recovery mechanism. PMID- 12740728 TI - Changes in finger coordination and responses to single pulse TMS of motor cortex during practice of a multifinger force production task. AB - We investigated the changes in finger coordination and in finger force responses to transcranial magnetic stimulation (TMS) applied over the motor cortex associated with a single practice session of an accurate ramp force production task. Subjects pressed with their index, middle and ring fingers onto three force transducers fixed to a rigid platform that was balanced on a narrow pivot under the middle finger. The task was to produce a smoothly increasing ramp of total force from 0 to 25 N over 4 s following a visual target. Subjects performed three brief series of trials without TMS (12 trials each) in the beginning, in the middle, and in the end of the experiment. The main part of the experiment involved 173 trials, and in each of them at random times in the ramp a suprathreshold TMS pulse was applied over the hand area of the contralateral motor cortex in order to evoke a twitch in the finger flexor muscles. At the end of the experiment the subjects also performed 12 constant force production trials, and TMS was unexpectedly applied in each trial. During the ramp force trials the amplitude of the response to TMS was largely independent of the force exerted at the time of stimulation, whereas in static holding trials the amplitude of the response increased with higher levels of background contraction. Over time subjects improved their overall tracking performance: the variance of the force trajectory (VarF(TOT)), as computed over sets of unperturbed trials, declined by 60% after the first 100 trials, but there was little additional improvement after the second 100 trials. Variance in the force finger space related to the total moment with respect to the pivot also showed a decline during the first half of practice and minimal further changes during the second half. In contrast, finger force variance that did not affect either total force or total moment showed no changes after the first 100 trials and a decline during the second 100 trials. This variance component quantified per finger was significantly larger than those related to the total force and total moment. The mean size of the TMS-induced phasic force increment decreased by 12% over the course of the 200 trials. The forces evoked in the index and ring fingers gradually became more equal, reducing the total moment with respect to the pivot and improving balance. We speculate that development of a relatively low twitch force with low total moment on the pivot made it easier for subjects to continue tracking after the TMS pulse. Such changes could well be correlated with the degree of corticospinal involvement in the task. The results suggest task specific, practice-related plastic changes in neural structures involved in the responses to TMS. PMID- 12740729 TI - Identification and functional analysis of six mycolyltransferase genes of Corynebacterium glutamicum ATCC 13032: the genes cop1, cmt1, and cmt2 can replace each other in the synthesis of trehalose dicorynomycolate, a component of the mycolic acid layer of the cell envelope. AB - By data mining in the sequence of the Corynebacterium glutamicum ATCC 13032 genome, six putative mycolyltransferase genes were identified that code for proteins with similarity to the N-terminal domain of the mycolic acid transferase PS1 of the related C. glutamicum strain ATCC 17965. The genes identified were designated cop1, cmt1, cmt2, cmt3, cmt4, and cmt5 ( cmt from corynebacterium mycolyl transferases). cop1 encodes a protein of 657 amino acids, which is larger than the proteins encoded by the cmt genes with 365, 341, 483, 483, and 411 amino acids. Using bioinformatics tools, it was shown that all six gene products are equipped with signal peptides and esterase domains. Proteome analyses of the cell envelope of C. glutamicum ATCC 13032 resulted in identification of the proteins Cop1, Cmt1, Cmt2, and Cmt4. All six mycolyltransferase genes were used for mutational analysis. cmt4 could not be mutated and is considered to be essential. cop1 was found to play an additional role in cell shape formation. A triple mutant carrying mutations in cop1, cmt1, and cmt2 aggregated when cultivated in MM1 liquid medium. This mutant was also no longer able to synthesize trehalose di coryno mycolate (TDCM). Since single and double mutants of the genes cop1, cmt1, and cmt2 could form TDCM, it is concluded that the three genes, cop1, cmt1, and cmt2, are involved in TDCM biosynthesis. The presence of the putative esterase domain makes it highly possible that cop1, cmt1, and cmt2 encode enzymes synthesizing TDCM from trehalose monocorynomycolate. PMID- 12740730 TI - Neuromuscular behaviour of the triceps surae muscle-tendon complex during running and jumping. AB - The present study examined the behaviour of the Achilles tendon (AT) - triceps surae (TS) muscle complex during running and long jump take-off. High AT forces were measured in the push-off phase in running even with very low EMG activity. In the long jump, high rate of stiffness development was a characteristic of the braking phase of the jump. The results suggest that high and well-coordinated activation patterns of the leg extensor muscles during the preactivation and eccentric phases together with high stretching velocities of muscle-tendon complex provide basis for appropriate tendomuscular stiffness. This together with high force at the end of the eccentric phase enables an effective push-off (concentric) phase. PMID- 12740731 TI - Validity and reliability of the Polar S710 mobile cycling powermeter. AB - The purpose of this study was to determine the validity and reliability of a new mobile bike-powermeter, Polar S710, in laboratory and field conditions, against the SRM crankset. Eight trained subjects performed in a random order six uphill cycling trials of 6-min duration at three different intensities (60 %, 75 % and 90 % of peak power output [PPO]). In addition, 44 other cyclists performed in the laboratory three cycling bouts each of 5-min duration at three different pedal cadences (60, 90 and 110 rpm) at the same absolute intensity (approximately 150 W). Bias between the two devices was correlated (r = 0.79) with the mean power in field conditions; with the S710 reading higher (p < 0.001) by 7.4 +/- 5.1 % than the SRM in the range of power studied. In other words, the mean differences between the two devices increased as the exercise intensity increased. The mean power output obtained with S710 was significantly higher (p < 0.001) by 6.8 +/- 7.9 W (bias x divided-by random error = 1.042 x divided-by 1.049) than the power obtained with SRM in laboratory conditions. Ninety-five percent of the differences of power measured with the S710 ranged between 21.4 W above to 8.3 W below the SRM in laboratory conditions. Mean differences between the two devices increased as the pedalling cadence increased (0.6 +/- 3.8 %, 4.4 +/- 3.7 % and 7.8 +/- 4.4 % at cadence of 60, 90 and 110 rpm respectively). Coefficients of variation in mean power across the four field-based trials at 75 % PPO was 2.2 % and 1.9 % for S710 and SRM, respectively. In conclusion, the S710 recorded power outputs higher than the SRM system in both field and laboratory conditions. Pedalling cadence and exercise intensity influenced differences in mean power. These characteristics make S710 a useful device for recreational cyclists but not for elite cyclists or scientists who require a greater accuracy and validity. However, the limits of the present study (short-term duration testing; single tested variables as intensity, posture, pedalling cadence) require further investigation for generalizing the present results to extensive use in "real world" cycling. PMID- 12740732 TI - Ventilatory and metabolic response to rebreathing the expired air in the snorkel. AB - The snorkel, which allows swimmers to keep their face down in the water while breathing, is widely used by divers, spear fishermen and monofin swimmers. A snorkel adds an additional dead space of 160-170 ml and causes an increase in the concentration of CO2 in the inspired gas due to expired air trapped in the snorkel which is then re-inspired. In this study the metabolic and the ventilatory response to rebreathing the expired air in the snorkel were investigated in twelve human subjects. A 2900 C Sensor Medics gas analyzer was used in breath-by-breath mode for the measurements. Ventilation (VE), respiratory rate (RR), tidal volume (TV), oxygen consumption (.VO 2) and carbon dioxide production (.VCO 2) were measured at rest and during light exercise both with and without the snorkel dead space. We observed a significant increase in all variables except RR, when subjects rebreathed the gas in the snorkel. The increase in ventilation resulted from an increase in tidal volume rather than increasing respiratory rate. We conclude that the work of breathing is increased when CO2 concentration is high in inspired gas and re-breathing while snorkelling can be prevented by a new snorkel design with a low-resistance two-way non rebreathing valve, which will allow the expired air flow into the water. PMID- 12740733 TI - The response of trained athletes to six weeks of endurance training in hypoxia or normoxia. AB - This study was performed to investigate the effect of training under simulated hypoxic conditions. Hypoxia training was integrated into the normal training schedule of 12 endurance trained cyclists. Athletes were randomly assigned to two groups and performed three additional training bouts per week for six weeks on a bicycle ergometer. One group (HG) trained at the anaerobic threshold under hypoxic conditions (corresponding to an altitude of 3200 m) while the control group (NG) trained at the same relative intensity at 560 m. Preceding and following the six training weeks, performance tests were performed under normoxic and hypoxic conditions. Normoxic and hypoxic .VO2max, maximal power output as well as hypoxic work-capacity were not improved after the training period. Testing under hypoxic conditions revealed a significant increase in oxygen saturation (SpO 2, from 67.1 +/- 2.3 % to 70.0 +/- 1.7 %) and in maximal blood lactate concentration (from 7.0 to 9.1 mM) in HG only. Ferritin levels were decreased from 67.4 +/- 16.3 to 42.2 +/- 9.5 microg/l (p < 0.05) in the HG and from 54.3 +/- 6.9 to 31.4+/- 8.0 microg/l (p = 0.17) in the NG. Reticulocytes were significantly increased in both groups by a factor of two. In conclusion, the integration of six weeks of high intensity endurance training did not lead to improved performance in endurance trained athletes whether this training was carried out in hypoxic or normoxic conditions. PMID- 12740734 TI - Heart-rate recommendations: transfer between running and cycling exercise? AB - With the expanding use of portable heart rate (HR) monitors in endurance sports, HR is increasingly used as a marker for exercise intensity. Hereby, HR at the so called individual anaerobic threshold (IAT) is one possible reference point. However, once determined, it is often attempted to apply HR recommendations from one type of ergometry to different kinds of exercises. We examined whether HR at IAT and at 4 mmol x l -1 blood lactate is predictable from cycling to running and vice versa. Data of 371 subjects (304 male, 67 female) were analyzed. All subjects underwent an incremental test on a treadmill (TR, starting speed 6 or 8 km x h -1, increments 2 km x h -1 every 3 min) and on a bicycle ergometer (BE, start at 50 Watt, increments 25 or 50 Watt every 3 min). IAT was determined at a net increase of lactate concentration of 1 - 5 mmol x l -1 above lactate concentration at lactate threshold for running (as in: Med Sci Sports Exerc 1998, 30 (10); 1552 - 1557) and 1.0 mmol x l -1 for cycling. A maximum time span of three weeks was allowed between the tests. We found that heart rate at IAT or at 4 mmol x l -1 blood lactate did not correlate between cycling and running. A sports specific test seems to be a prerequisite for reliable heart rate recommendations. PMID- 12740735 TI - Effects of active recovery under a decreasing work load following intense muscular exercise on intramuscular energy metabolism. AB - The effect of active recovery at a decreasing % of MVC following intense muscular exercise on intramuscular pH was investigated in vivo using 31P-MRS. Seven healthy men participated, and their right wrist flexor muscle group was examined. The subjects were asked to flex their right wrist at 60 % of the maximum voluntary contraction (MVC) every 2 s until the intracellular pH in the wrist flexor muscle decreased to 6.4. After the exercise period, the subjects underwent active or passive recovery for 10 min. For the active recovery (AR), the subject was asked to continue exercising at a decreasing % of MVC, decreasing from 25 to 5 % MVC every two min during AR. 31P-MR-spectra were obtained throughout the experiments, and from the spectra the intracellular pH (pHi) was calculated as an indicator of intracellular events. AR data were compared to data collected during passive recovery (PR). During AR, the pHi increased immediately after the exercise period; whereas in that of PR, it did not recover within 5 minutes after exercise. The results suggested that mild exercise was an effective manoeuver to promote recovery from intramuscular metabolic acidosis. PMID- 12740736 TI - 31P-MRS characterization of sprint and endurance trained athletes. AB - Muscle metabolism and force production were studied in sprint trained runners, endurance trained runners and in untrained subjects, using 31P-MRS. 31P-spectra were obtained at a time resolution of 5 s during four maximal isometric contractions of 30-sec duration, interspersed by 60-sec recovery intervals. Resting CrP/ATP ratio averaged 3.3 +/- 0.3, with no difference among the three groups. The sprint trained subjects showed about 20 % larger contraction forces in contraction bouts 1 and 2 (p < 0.05). The groups differed with respect to CrP breakdown (p < 0.05), with sprinters demonstrating about 75 % breakdown in each contraction compared to about 60 % and 40 % for untrained and endurance trained subjects, respectively (p < 0.05). The endurance trained runners showed almost twice as fast CrP recovery (t 1/2 = 12.5 +/- 1.5) compared to sprint trained (t 1/2 = 22.5 +/- 2.53) and untrained subjects (t 1/2 = 26.4 +/- 2.8). From the initial rate of CrP resynthesis the rate of maximal aerobic ATP synthesis was estimated to 0.74 +/- 0.07, 0.73 +/- 0.10 and 0.33 +/- 0.07 mmol ATP x kg -1 wet muscle x sec -1 for sprint trained, endurance trained and untrained subjects, respectively. Only the sprint trained and the untrained subjects displayed a significant drop in pH and only during the first of the four contractions, about 0.2 and 0.1 pH units, respectively, indicating that only under those contractions was the glycolytic proton production larger than the proton consumption by the CK reaction. Also, in the first contraction the energy cost of contraction was higher for the sprinters compared to the two other groups. The simple 31P-MRS protocol used in the present study demonstrates marked differences in force production, aerobic as well as anaerobic muscle metabolism, clearly allowing differentiation between endurance trained, sprint trained and untrained subjects. PMID- 12740737 TI - Alteration of neuromuscular function after a prolonged road cycling race. AB - The aim of this study was to characterize neuromuscular fatigue in knee extensor muscles after a prolonged cycling exercise. During the two days preceding a 140 km race (mean +/- SD duration: 278.2 +/- 24.9 min) and 15 to 30 min after, maximal percutaneous electrical stimulations were applied to the femoral nerve of 11 trained cyclists. Electrically evoked superimposed twitches and trains of 6 stimulations were delivered during isometric maximal voluntary contraction (MVC) to determine maximal voluntary activation (% VA). Knee extensors MVC decreased with fatigue from 158.2 +/- 29.6 to 144.2 +/- 30.0 Nm (p < 0.01), but no central activation failure was detected after the race. The average rate of twitch force development increased significantly from 414 +/- 106 to 466 +/- 102 N x m x s-1 (p < 0.05) and a tendency toward higher peak twitch tension (p = 0.052) was found in the fatigued state. Short tetanus at 20 Hz and 80 Hz were also applied to 4 cyclists, but these fused and unfused tetanic forces were not significantly modified with fatigue. From these results, it can be concluded that the small but significant isometric strength loss measured less than 30 min after the end of a long distance road cycling race is not due to central fatigue. It is also suggested that a raise in peak twitch tension is not necessarily associated with enhanced neuromuscular function. PMID- 12740738 TI - Androgenic anabolic steroid use and severe hypothalamic-pituitary dysfunction: a case study. AB - The data of the present case demonstrate that the abuse of androgenic anabolic steroids (AAS) may lead to serious health effects. Although most clinical attention is usually directed towards peripheral side effects, the most serious central side effect, hypothalamic-pituitary-dysfunction, is often overlooked in severe cases. Although this latter central side-effect usually recovers spontaneously when AAS intake is discontinued, the present case shows that spontaneous recovery does not always take place. We suggest that hypothalamic pituitary dysfunction should always be considered in the differential diagnosis in athletes seen with typical presentation of anabolic steroid use. In order to regain normal hypothalamic-pituitary function, supraphysiological doses of 200 microg LH-RH should be considered when the physiological challenge test with LH RH (50 microg) fails to show an acceptable response. PMID- 12740739 TI - Urinary profile of androgen metabolites in a population of sportswomen during the menstrual cycle. AB - The aim of this investigation was to evaluate the urinary profile of androgen metabolites during menstrual cycle in both young-trained female athletes, and young sedentary women, not presenting any pathological signs. Urines were collected for 24 hours (08 : 00 a. m. the first day to 08 : 00 a. m. the second day) from all sportive and sedentary subjects. All steroids were measured by specific radioimmunological analysis, and the implications of these results in terms of concentrations and modifications by exercise will be discussed. During follicular phase, control values were respectively, testosterone glucuronide (TG): 1.67 +/- 0.70 nmol x mmol C -1; epitestosterone glucuronide (ETG): 2.51 +/- 0.88 nmol x mmol C -1; TG/ETG ratio: 0.72 +/- 0.26 and cortisol (FLU): 10.02 +/- 0.79 nmol x mmol C -1. No significant modifications were observed during luteal phase (respectively: TG: 1.48 +/- 0.50 nmol x mmol C -1; ETG: 2.65 +/- 0.93 nmol x mmol C -1; TG/ETG ratio: 0.67 +/- 0.31 and FLU: 9.29 +/- 3.37 nmol x mmol C -1. Similarly, no significant effect of physical training was observed on studied parameters between these two groups during either follicular phase (TG: 1.96 +/- 1.00 nmol x mmol C -1; ETG: 1.97 +/- 0.70 nmol x mmol C -1; TG/ETG ratio: 0.66 +/ 0.05 and FLU: 11.31 +/- 3.73 nmol x mmol C -1) or luteal phase (TG: 1.93 +/- 0.86 nmol x mmol C -1; ETG: 3.19 +/- 1.23 nmol x mmol C -1; TG/ETG ratio: 0.69 +/ 0.33 and FLU: 9.52 +/- 3.86 nmol x mmol C -1). It is concluded that although physical training could play a role in androgen metabolism, it has no significant incidence on urinary TG/ETG ratio. This study thus confirms that sportswomen can also be considered as normal subjects when they do not present any obvious endocrine disorder induced by physical activity. PMID- 12740740 TI - Fiber type characteristics and myosin light chain expression in a world champion shot putter. AB - Muscle biopsies from m. vastus lateralis of two world class shot putters (shot putter 1 and 2) and the untrained brother of shot putter 1 were analyzed for fiber type distribution with ATPase staining and in situ hybridization for the expression of alkali myosin light chain (MLC) isoforms. Shot putter 2 had a predominance of type II fibers (67 X) and distinct hypertrophy of type I as well as type II fibers (fiber areas of 5939 and 8531 microm2). In shot putter 1, type II fibers amounted to only 40%, due to their selective hypertrophy, however type II fibers (10265 microm2) accounted for 67 2% of the total cross-sectional area. The type I fibers in shot putter 1 were similar in size to his untrained brother (3430 vs 3790 microm2). After 3 years of active detraining, type II fibers of shot putter 1 had reduced in size to values closer to those of his brother (7746 and 6340 microm2). The large difference between type I and type II fiber size, even in the untrained state, in both shot putter 1 and his brother is not usually seen in humans and maybe a genetic characteristic. We suggest that the ability to selectively increase the relative area of his type II fibers in the 15 years of strength training was a key element in his success as a shot putter. The observed increase in the expression of fast myosin light chain mRNAs in both fiber types is indicative of further adjustment of the myofibrillar apparatus towards the generation of very high peak power. PMID- 12740741 TI - Involvement of endogenous growth hormone-releasing hormone (GHRH) in the exercise related response of growth hormone. AB - The aim of this study was to investigate the involvement of endogenous growth hormone-releasing hormone (GHRH) in the growth hormone (GH) release during strenuous exercise (EX). Eight healthy male subjects (age: 22.1 +/- 0.8 yr, body mass index: 22.2 +/- 0.9 kg/m 2, .VO 2 max: 52.2 +/- 0.5 ml/min/kg [mean +/- SEM]) were exposed to incremental EX until volitional exhaustion (cycle ergometry), and in random order to a maximally stimulating bolus injection of 100 microg GHRH, or to combined administration of 100 microg GHRH and EX (GHRH+EX). Serial blood samples in the fasted state were taken immediately before the start of each trial, and at appropriate intervals over 2 h. Total GH availability was calculated as area under the response curve (AUC), corrected for differences in baseline values. The results showed that peak serum GH levels to GHRH alone and EX alone were not significantly different: 41.5 +/- 9.0 microg/l and 64.1 +/- 8.1(mean +/- SEM). Peak GH level to GHRH+EX was 156.1 +/- 19.9 microg/l, which was significantly greater than to either stimulus alone (p < 0.02) or additively (105.6 +/- 17.1 microg/l, p < 0.02). AUC's to GHRH alone and EX alone were not significantly different (3242 +/- 839 vs. 2472 +/- 408 microg/l x 120 min). AUC to GHRH+EX (7807 +/- 1221 microg/l x 120 min) was greater than to either stimulus alone (p < 0.02) or additively (5714 +/- 1247 microg/l x 120 min, p < 0.02). This indicates a potentiating (synergistic) effect between GHRH and EX. We postulate that GH responses to strenuous EX are only partially due to maximal GHRH activation. Next to complete inhibition of hypothalamic somatostatin activity, which is achieved by strenuous exercise, activation of endogenous GH-releasing peptides, such as Ghrelin, must be operative. PMID- 12740742 TI - Plasma zinc and immune markers in runners in response to a moderate increase in training volume. AB - Changes in plasma zinc concentration and markers of immune function were examined in a group of 10 male runners (n = 10) following a moderate increase in training over four weeks. Seven sedentary males acted as controls. Fasting blood samples were taken at rest, before (T0) and after (T4) four weeks of increased (+ 16 %) training and after two weeks of reduced (-31 %) training (T6). Blood was analysed for plasma zinc concentration, differential leucocyte counts, lymphocyte subpopulations and lymphocyte proliferation using incorporation of 3H-thymidine. The runners increased their training volume by 16 % over the four weeks. When compared with the nonathletes, the runners had lower concentrations of plasma zinc (p = 0.012), CD3 + (p = 0.042) and CD19 + lymphocytes (p = 0.010) over the four weeks. Lymphocyte proliferation in response to Concanavalin A stimulation was greater in the runners (p = 0.0090). Plasma zinc concentration and immune markers remained constant during the study. Plasma zinc concentration correlated with total leucocyte counts in the athletes at T6 (r = -0.72, p < 0.05) and with Pokeweed mitogen stimulation in the nonathletes at T6 (r = -0.92, p < 0.05). Therefore, athletes are unlikely to benefit from zinc supplementation during periods of moderately increased training volume. PMID- 12740743 TI - Influence of iron status and iron supplements on natural killer cell activity in trained women runners. AB - Twenty-two trained women runners (.VO2peak 48.1 + 1.2 ml x kg -1 x min -1) were divided into an iron supplement (n = 13) or placebo group (n = 9) based on initial serum ferritin concentration (24.2 +/- 2.9 and 58.5 +/- 4.0 microg x l 1, respectively). Exercise consisted of a 35-min run (80 % .VO2peak) and was performed at week 0 (WK0), after two weeks of intensified training (WK2) and after eight weeks recovery training (WK10). The eight weeks recovery training were concomitant with subjects taking iron supplements or placebo in a double blind fashion. Concentrations of serum ferritin, serum iron and total iron binding capacity were assessed pre-exercise and complete blood count, natural killer cell activity (NKACT), and cell surface markers for CD3+, CD4+, CD3+,CD8+, CD3-, CD16+, CD56+ cells were determined both pre- and post-exercise. Serum ferritin concentrations were significantly (p < 0.05) increased on WK10 compared to WK2 (time effect). NKACT (%lysis) and NK cell number was lower (p < 0.05) at WK0 for supplement (42.9 +/- 1.9 % and 305.5 +/- 15.0 x 10 6 x l -1, respectively) compared to placebo groups (50.9 +/- 2.0 and 406.1 +/- 25.6, respectively). Two weeks of intensified training did not alter indices of host defense. In conclusion, NKACT and NK cell numbers were lower in subjects with greater body mass and lower iron stores (p < 0.05), but were not significantly altered after two weeks of intensified training or when serum ferritin levels increased. PMID- 12740744 TI - Tennis, incidence of URTI and salivary IgA. AB - Tennis played at an elite level requires intensive training characterized by repeated bouts of brief intermittent high intensity exercise over relatively long periods of time (1 - 3 h or more). Competition can place additional stress on players. The purpose of this study was to investigate the temporal association between specific components of tennis training and competition, the incidence of upper respiratory tract infections (URTI), and salivary IgA, in a cohort of seventeen elite female tennis players. Timed, whole unstimulated saliva samples were collected before and after selected 1-h training sessions at 2 weekly intervals, over 12 weeks. Salivary IgA concentration was measured by ELISA and IgA secretion rate calculated (microg IgA x ml -1 x ml saliva x min -1). Players reported URTI symptoms and recorded training and competition in daily logs. Data analysis showed that higher incidence of URTI was significantly associated with increased training duration and load, and competition level, on a weekly basis. Salivary IgA secretion rate (S-IgA) dropped significantly after 1 hour of tennis play. Over the 12-week period, pre-exercise salivary IgA concentration and secretion rate were directly associated with the amount of training undertaken during the previous day and week (p < 0.05). However, the decline in S-IgA after 1 h of intense tennis play was also positively related to the duration and load of training undertaken during the previous day and week (p < 0.05). Although exercise-induced suppression of salivary IgA may be a risk factor, it could not accurately predict the occurrence of URTI in this cohort of athletes. PMID- 12740746 TI - [Projective identification and denial of pregnancy--considerations of the reasons and background of unrecognized pregnancy also undiagnosed by a physician]. AB - BACKGROUND: In denial of pregnancy, the pregnant woman does not consciously perceive the pregnancy and, in extreme cases, awareness occurs only during delivery. The attending physicians also often fail to recognize the pregnancy, even though the somatic complaints leading to consultation with a physician are typically pregnancy associated. This "iatrogenic participation" was described in the earliest historical publications. Different theories are presented in this paper to elaborate this phenomenon. Elementary deficiencies in perception or incompetence do not explain most cases. PATIENTS AND METHODS: Twenty five women with denial of pregnancy, interviews PSYCHODYNAMIC EXPLANATIONS: Based on the deep-rooted subjective attitude of not being pregnant, the pregnant woman is able to include family, friends, and associates into the denial of pregnancy mindset. In a similar way, she is able to influence her doctor. The woman's autosuggestive wishful notions of not being pregnant receive suggestive confirmation by the physician's misdiagnosis and lead to a continuing denial of pregnancy. PROJECTIVE IDENTIFICATION: In 1946, M. Klein introduced this term to describe a certain defense mechanism relating to fantasies and accompanying object relationships. The self initially successfully disposes of unwanted aspects, splits them off, and transfers them to another person, to finally reclaim them in a modified form. Certain elements of this psychoanalytical concept can characterize the interaction between physician and pregnant woman, which, in the case of denied pregnancies, does not lead to a diagnosis of pregnancy. Through projection, the pregnant woman is capable of manipulating the physician so that he perceives her, according to her wishes, as not being pregnant and misdiagnoses her correspondingly as "service in return". Opportunities for more mature handling of the denied content in terms of psychological development through accurate diagnosis of the pregnancy are indicated. PMID- 12740747 TI - [Cerebral hemorrhage and periventricular leucomalacia in preterm neonates after intravenous tocolysis with fenoterol: results of postnatal ultrasound examination]. AB - BACKGROUND: This study tested whether tocolysis with beta-adrenergic agonists (Fenoterol) had an effect on the frequency of cerebral lesions in preterm neonates. PATIENTS AND METHODS: Head ultrasound scans of preterm neonates who were born after long-term (> 24 h) tocolysis were compared with scans of preterm neonates without preceding tocolysis. The gestational and neonatal data were analyzed retrospectively. RESULTS: Preterm neonates after (n = 102) and without (n = 101) tocolysis were subdivided into three groups according to their gestational age (23 - 28 wk: n = 41; 29 - 33 wk: n = 66; 34 - 36 wk: n = 96). Within these groups, no significant differences were found with respect to birth weight, rate of cesarean section, or pulmonary morbidity. Preterm babies < 28 weeks of gestation from the control group had lower Apgar scores (after 1 and 5 minutes, respectively) and arterial umbilical cord pH values. Intravenous tocolysis did not lead to an increase in pseudocystic periventricular leucomalacia (PVL) or intracerebral hemorrhage (ICH) in any of the subgroups studies. However, cerebral lesions were found in preterm neonates after tocolysis who exhibited signs of infection (29 - 33 wk: PVL n = 2; 23 - 28 wk: ICH n = 1) and in preterm neonates without tocolysis who had undergone fetal hypoxia or abruptio placentae (29 - 33 wk: PVL n = 4; antenatal terminal vein bleeding n = 1; 23 - 28 wk: PVL n = 2; terminal vein bleeding n = 5; posterior cerebral artery bleeding n = 1). When compared to preterm neonates of 34 - 36 weeks of gestation, the risk of infection was increased 4-fold in neonates of 29 - 33 weeks of gestation (odds ratio 5.43, 1.10 - 26.83) and 10-fold in neonates of 23 - 28 weeks of gestation (odds ratio 20.50, 3.65 - 115.03). Chorioamnionitis also was a more common finding in preterm neonates < 28 weeks of gestation. CONCLUSION: Preterm neonates who were born after intravenous long-term (> 24 h) tocolysis with Fenoterol do not exhibit an increase in periventricular leucomalacia or intracranial hemorrhage. The occurrence of cerebral lesions in these patients merely depends on their degree of immaturity and on the presence or absence of perinatal infection. In preterm neonates without tocolysis, brain lesions are mainly associated with hypoxic events. PMID- 12740745 TI - [Surfactant proteins A and D: major factors of the immune response of the lung]. AB - Surfactant proteins (SP) A and D play a key role in pulmonary host defense of preterm and term newborns. SP-A and SP-D have structural elements of both collagens and lectins, for which they are called "collectins". SP-A and SP-D coordinate the innate and adaptive immune response in the lung and convey protection against infection in amniotic fluid and vernix caseosa. They are pattern-recognizing molecules of the innate immune system that are involved in binding, agglutination of pathogens, chemotaxis of neutrophils, phagocytosis of pathogens, and production of reactive oxygen species. In addition, SP-A and SP-D interact with the adaptive immune system by reducing interleukin-2 production and T lymphocyte proliferation. PMID- 12740748 TI - [Intact cervical pregnancy]. AB - The authors describe a case of intact cervical pregnancy in a 24-year-old secundigravida. The patient was treated successfully with Methotrexate. Conservative treatment is the first choice in the therapy of uncomplicated cervical pregnancy. Conservative and operative therapeutic procedures are discussed. PMID- 12740749 TI - [Family planning and pregnancy with thalassemia major]. AB - We present the case of a young Italian woman with thalassemia major and describe a series of complications such as infertility, fetal growth restriction, difficult intubation, and cardiac decompensation that occurred in the course of family planning, pregnancy, and the puerperal period. By review of the literature and based on own experiences, we show how the preconceptional consultation and the care during a pregnancy, especially the individualized planning of delivery, can be optimized. The intensive and interdisciplinary care by obstetricians and internists is very important. PMID- 12740751 TI - ["Recent advances in neonatal medicine", 18-20 October 2002, Wurzburg, Germany]. PMID- 12740750 TI - [Hodgkin's disease in pregnancy--case report and literature review]. AB - We report the case of a 24-year-old Gravida II/Para I who was diagnosed with Hodgkin's disease (stage 2A) at week 28 of pregnancy. Due to clinical progression, a dose-reduced adriamycin, bleomycin, vincristin, dacarbazine chemotherapy was started at week 32. After unsuccessful induction of labor at week 35, secondary cesarean section with a healthy newborn was performed. Chemotherapy was continued with full dose 10 days postpartum. We discuss clinical management as well as diagnostic and therapeutic options with respect to the available literature. PMID- 12740752 TI - [DRGs in neurology and psychiatry--progress or risk?]. PMID- 12740754 TI - [The development of a case group concept for patients with mental diseases]. AB - In the year 2000 the German legislative decided to implement a case-dependent reimbursement system in an acute inpatient setting beginning in 2003. The German Diagnoses Related Group System (G-DRGs) will adopt the refined version of the Australian-Diagnoses Related Group System (AR-DRG, version 4.1). Based on empirical data and experts panels the acute psychiatric hospitals have been exempted from the adoption of the case-based payment system. With regard to national and international experiences in the development of patient classification systems in psychiatry and medical rehabilitation this article illustrates the development of a case group concept for patients with mental disorders. PMID- 12740753 TI - [DRGs and Psych-PV]. PMID- 12740755 TI - [Placebo-controlled antidepressant drug trials. Considering the ethical implications]. AB - For the approval of a new antidepressant for clinical use drug authorities still demand the proof of a significantly superior efficacy of the compound over placebo shown in controlled comparison studies. On the other side serious ethical concerns have been raised upon the use of placebos in clinical trials with depressed patients in that this would implicitly mean a withholding of an efficacious treatment from patients in the control groups of such trials. This article summarises frequently quoted arguments for and against placebo use in antidepressant clinical trials focussing on the specific issues involved in the therapeutic effect of antidepressants and placebos in depressed patients and discusses measures that might contribute to study designs that may better evaluate the true efficacy of a new drug while keeping the risk for the individual patient at the lowest possible level. A worldwide solution of the problem is clearly called for. A ban of placebos in antidepressant trials as it might be derived from the Declaration of Helsinki should be postponed until antidepressants with a more convincing superiority over placebo have been developed. PMID- 12740756 TI - [Initial use of risperidone in the treatment of acutely exacerbated schizophrenic patients--an interim analysis]. AB - OBJECTIVE: The effectiveness of atypical antipsychotic agents in the treatment of acute schizophrenic episodes is still a subject of controversial debate. The objective, therefore, was to investigate the efficacy and tolerability of an initial therapy with the atypical antipsychotic agent risperidone in acutely exacerbated patients under the conditions of clinical practice. A sub-analysis was performed to show if highly agitated and aggressive patients may profit from an initial risperidone therapy as well. MATERIAL AND METHODS: In a still ongoing prospective multicentre observational trial, schizophrenic patients with acute exacerbations treated with risperidone within 24 hours of in-patient admission were observed for six weeks. Patients showing a total score of > or = 15 in the items "excitement", "hostility" and "uncooperativeness" of the Positive and Negative Syndrome Scale (PANSS) were defined as highly agitated patients. Evaluation of efficacy was carried out according to a modified PANSS, the Clinical Global Impression (CGI) and the Brief Psychiatric Rating Scale (BPRS). RESULTS: 1,117 patients were evaluated. An improvement of all parameters was shown in the whole study group (51 % males, age 39.8 + 14.3 years, paranoid schizophrenia in 70.1 % of cases) and in particular in the subgroup of highly agitated patients. In these patients (n = 163), a greater improvement of symptoms was observed. Only in 4,1 % of cases was risperidone discontinued because of side effects. At the end of the observation, the mean dosage was 5.1 mg/day in both groups. More than 50 % of the patients were finally treated with a risperidone monotherapy. CONCLUSION: The initial acute treatment with risperidone proves to be effective and safe even for highly agitated schizophrenic patients under the conditions of clinical practice. PMID- 12740757 TI - [Oxcarbazepine in the treatment of affective and schizoaffective disorders]. AB - The therapeutic value of anticonvulsants in affective and schizoaffective disorders was documented in several clinical trials. Oxcarbazepine (OXC), a keto derivative of carbamazepine, which appears to have a preferable side effect profile compared to carbamazepine, has also shown antimanic efficacy in affective and schizoaffective disorders in clinical studies since the early 80's, but was not further investigated regarding these indications. Therefore, the value of OXC in the treatment of affective and schizoaffective disorders requires evaluation. Literature was reviewed with regard to pharmacokinetic and pharmacodynamic characteristics of OXC, drug-drug interactions relevant in pharmacopsychiatry, and clinical effects in these disorders. According to the literature OXC is regarded effective in acute mania and appears to allow reduction of the neuroleptic medication required for the treatment of affective and schizoaffective disorders. In addition, it has a preferable pharmacokinetic profile with less severe side effects compared to other anticonvulsants and neuroleptics. Furthermore, it appears to be well tolerated if augmented to neuroleptics or antidepressants, since OXC does not interact substantially with the cytochrome P450-enzyme-system. However, despite promising effects of OXC, few clinical studies have been published in the last 16 years. We conclude that further studies should validate the antimanic efficacy of OXC and evaluate possible pharmacopsychiatric indications as well as limitations of this psychotropic compound. PMID- 12740758 TI - [Patients' perspective on the quality of informed consent into psychopharmacological treatment in schizophrenia and depression]. AB - BACKGROUND: Respect for autonomy and the right to self-determination require informed consent of patients before any medical procedure. So far the majority of empirical studies on informed consent deal mainly with single aspects e. g. delivery and recall of information and legal questions from a physicians' perspective. In our study we examine the quality of informed consent to psychopharmacological treatment from the perspective of patients with schizophrenia and depression in a German university department of psychiatry. METHODS: Face to face interviews using a structured questionnaire by a research psychiatrist. The participants taking part in this study were inpatients diagnosed with schizophrenia and depression. All patients received medication for their condition. RESULTS: Half of the patients rated the information they received and the process of informed consent positive. Only a minority recalls information regarding the aim and the risks of the treatment. 20 % indicated that they have been informed about alternative treatment options. A quarter of the patients did not know that they had a right to refuse any treatment. CONCLUSION: Compared with studies including patients with somatic illnesses the participants in our study judge the process of informed consent rather critical. The results need to be discussed in the light of the special aspects of psychopharmacological treatment and different cultural and institutional settings. PMID- 12740759 TI - [Age dependence of Doppler parameters in the basal cerebral arteries evaluated by transcranial color-coded duplex sonography. Reference data from 290 volunteers]. AB - AIM: In this study the Doppler parameters of the basal intracranial arteries and the insonation in axial and coronary plane by temporal approach were established in healthy volunteers of all age groups with transcranial color-coded duplex sonography (TCDS). METHODS: 290 healthy probands (age 0 to 91 years) were investigated through the temporal and transnuchal ultrasound window. The angle corrected flow velocities and the resistance indices of the basal brain arteries were measured. RESULTS: The flow velocities rapidly increase in children (from 3 years up to puberty) and gradually decrease with increased age. Vice versa, the resistance indices decrease in children and increase in older people. The flow velocities show significant differences by axial and coronary insonation in certain arteries of the circle of Willis. CONCLUSION: The Doppler parameters of the basal intracranial arteries evaluated with TCDS depend from age. The flow velocities and resistance indices are similar in children and older people. The coronary transtemporal approach is considered a useful completion of the axial investigation. PMID- 12740760 TI - Definition of a critical region on chromosome 18 for congenital aural atresia by arrayCGH. AB - Deletions of the long arm of chromosome 18 occur in approximately 1 in 10,000 live births. Congenital aural atresia (CAA), or narrow external auditory canals, occurs in approximately 66% of all patients who have a terminal deletion 18q. The present report describes a series of 20 patients with CAA, of whom 18 had microscopically visible 18q deletions. The extent and nature of the chromosome-18 deletions were studied in detail by array-based comparative genomic hybridization (arrayCGH). High-resolution chromosome-18 profiles were obtained for all patients, and a critical region of 5 Mb that was deleted in all patients with CAA could be defined on 18q22.3-18q23. Therefore, this region can be considered as a candidate region for aural atresia. The array-based high-resolution copy-number screening enabled a refined cytogenetic diagnosis in 12 patients. Our approach appeared to be applicable to the detection of genetic mosaicisms and, in particular, to a detailed delineation of ring chromosomes. This study clearly demonstrates the power of the arrayCGH technology in high-resolution molecular karyotyping. Deletion and amplification mapping can now be performed at the submicroscopic level and will allow high-throughput definition of genomic regions harboring disease genes. PMID- 12740762 TI - A neutral explanation for the correlation of diversity with recombination rates in humans. AB - One of the most striking findings to emerge from the study of genomic patterns of variation is that regions with lower recombination rates tend to have lower levels of intraspecific diversity but not of interspecies divergence. This uncoupling of variation within and between species has been widely interpreted as evidence that natural selection shapes patterns of genetic variability genomewide. We revisited the relationship between diversity, divergence, and recombination in humans, using data from closely related species and better estimates of recombination rates than previously available. We show that regions that experience less recombination have reduced divergence to chimpanzee and to baboon, as well as lower levels of diversity. This observation suggests that mutation and recombination are associated processes in humans, so that the positive correlation between diversity and recombination may have a purely neutral explanation. Consistent with this hypothesis, diversity levels no longer increase significantly with recombination rates after correction for divergence to chimpanzee. PMID- 12740761 TI - Mutations in the transcription factor gene SOX18 underlie recessive and dominant forms of hypotrichosis-lymphedema-telangiectasia. AB - Hereditary lymphedema is a developmental disorder characterized by chronic swelling of the extremities due to dysfunction of the lymphatic vessels. Two responsible genes have been identified: the vascular endothelial growth factor receptor 3 (VEGFR3) gene, implicated in congenital lymphedema, or Milroy disease, and the forkhead-related transcription factor gene FOXC2, causing lymphedema distichiasis. We describe three families with an unusual association of hypotrichosis, lymphedema, and telangiectasia. Using microsatellite analysis, we first excluded both VEGFR3 and FOXC2 as causative genes; we then considered the murine ragged phenotype, caused by mutations in the Sox18 transcription factor, as a likely counterpart to the human disease, because it presents a combination of hair and cardiovascular anomalies, including symptoms of lymphatic dysfunction. Two of the families were consanguineous; in affected members of these families, we identified homozygous missense mutations in the SOX18 gene, located in 20q13. The two amino acid substitutions, W95R and A104P, affect conserved residues in the first alpha helix of the DNA-binding domain of the transcription factor. In the third family, the parents were nonconsanguineous, and both the affected child and his brother, who died in utero with hydrops fetalis, showed a heterozygous nonsense mutation that truncates the SOX18 protein in its transactivation domain; this substitution was not found in genomic DNA from either parent and hence constitutes a de novo germline mutation. Thus, we show that SOX18 mutations in humans cause both recessive and dominant hypotrichosis-lymphedema-telangiectasia, suggesting that, in addition to its established role in hair and blood vessel development, the SOX18 transcription factor plays a role in the development and/or maintenance of lymphatic vessels. PMID- 12740766 TI - Technical aspects of the Rastelli and atrial switch procedure for congenitally corrected transposition of the great arteries with ventricular septal defect and pulmonary stenosis or atresia: results of therapy. AB - In conventional surgery for the associated lesions of congenitally corrected transposition of the great arteries, the right ventricle remains in the systemic circulation. In this situation, the right ventricle and tricuspid valve fail in an unpredictable manner. The double switch procedure was introduced to restore the morphologic left ventricle to the systemic circulation and considerable success has been seen over the last 10 years with this approach. The Rastelli and atrial switch procedure can be applied to patients with congenitally corrected transposition of the great arteries and pulmonary stenosis or atresia and a suitably placed ventricular septal defect in the outlet septum of the ventricle beneath the aortic valve. Thus, the left ventricle can be restored to the systemic circulation. The Rastelli-atrial switch is a complex operative procedure, but the operative risk and long-term results are good without evidence in the mid-term of ventricular failure as has been associated with the conventional repair. A disadvantage is that these patients require valved conduit changes over the years. PMID- 12740767 TI - Technical aspects of the combined arterial switch and senning operation for congenitally corrected transposition of the great arteries. AB - Congenitally corrected transposition of the great arteries (ccTGA) is a rare defect characterized by discordant atrioventricular and ventriculoarterial connections. Symptoms result from one or a combination of associated cardiovascular malformations, including ventricular septal defect, pulmonary stenosis or atresia, and tricuspid valve dysfunction. Survival of patients with ccTGA is often determined by the associated anomalies, but even in isolation ccTGA results in a diminished life expectancy. Regurgitation of the tricuspid valve and decreased morphologic right ventricular function tend to develop and worsen over time. Anatomic repair by double switch technique may improve survival of patients with ccTGA by establishing the morphologic left ventricle in the systemic circulation. PMID- 12740763 TI - Identification of a novel gene and a common variant associated with uric acid nephrolithiasis in a Sardinian genetic isolate. AB - Uric acid nephrolithiasis (UAN) is a common disease with an established genetic component that presents a complex mode of inheritance. While studying an ancient founder population in Talana, a village in Sardinia, we recently identified a susceptibility locus of approximately 2.5 cM for UAN on 10q21-q22 in a relatively small sample that was carefully selected through genealogical information. To refine the critical region and to identify the susceptibility gene, we extended our analysis to severely affected subjects from the same village. We confirm the involvement of this region in UAN through identical-by-descent sharing and autozygosity mapping, and we refine the critical region to an interval of approximately 67 kb associated with UAN by linkage-disequilibrium mapping. After inspecting the genomic sequences available in public databases, we determined that a novel gene overlaps this interval. This gene is divided into 15 exons, spanning a region of approximately 300 kb and generating at least four different proteins (407, 333, 462, and 216 amino acids). Interestingly, the last isoform was completely included in the 67-kb associated interval. Computer-assisted analysis of this isoform revealed at least one membrane-spanning domain and several N- and O-glycosylation consensus sites at N-termini, suggesting that it could be an integral membrane protein. Mutational analysis shows that a coding nucleotide variant (Ala62Thr), causing a missense in exon 12, is in strong association with UAN (P=.0051). Moreover, Ala62Thr modifies predicted protein secondary structure, suggesting that it may have a role in UAN etiology. The present study underscores the value of our small, genealogically well characterized, isolated population as a model for the identification of susceptibility genes underlying complex diseases. Indeed, using a relatively small sample of affected and unaffected subjects, we identified a candidate gene for multifactorial UAN. PMID- 12740769 TI - The effect of pulmonary circulation hemodynamics on right ventricular unloading via the bidirectional Glenn shunt: implications for congenitally corrected transposition repair. AB - The bidirectional Glenn shunt has been successfully applied as an adjunct to ventricular septal defect closure and pulmonary valvulotomy to treat congenitally corrected transposition of the great arteries (ccTGA). The purpose of this study was to examine the volume and pressure unloading effects of the bidirectional Glenn shunt on the hypertrophied pulmonary ventricle in a canine model of ccTGA. Five beagles underwent survival surgery to band the pulmonary artery. Three months later, a polytetrafluoroethylene graft was anastomosed to the superior vena cava and right pulmonary artery. The graft or superior vena cava was clamped to create the normal or bidirectional Glenn circulation, and hemodynamic data were recorded. The bidirectional Glenn shunt significantly reduced right ventricular volume loading and stroke work. Dogs with normal pre-bidirectional Glenn cardiac outputs had greatly reduced right ventricular volumes and pressures with the bidirectional Glenn shunt. Dogs with pre-bidirectional Glenn right ventricular dysfunction had moderate volume but no pressure decreases with the bidirectional Glenn shunt owing to improved left ventricular output. In these dogs it is likely that the decreased level of pressure and volume unloading is because of a concomitant improvement in left ventricular output post bidirectional Glenn shunt placement. The bidirectional Glenn shunt is effective at unloading the right ventricle in a canine model of ccTGA. PMID- 12740768 TI - Physiologic versus anatomic repair of congenitally corrected transposition of the great arteries. AB - When biventricular repair of congenitally corrected transposition of the great arteries (ccTGA) is possible, complete anatomic (left ventricle to the aorta, right ventricle to the pulmonary artery, and associated atrial baffle operation) or physiologic (left ventricle to pulmonary artery, right ventricle to aorta) connections are complex and associated with significant long-term problems such as arrhythmias, subaortic obstruction, valvar dysfunction, ventricular-to pulmonary artery conduit changes, and deteriorating ventricular function. Recent advances have supported physiologic, anatomic, and single-ventricle repairs. The purpose of this article is to review the natural history of ccTGA, compare and contrast the physiologic and anatomic repairs, and reintroduce a new concept of physiologic repair for patients with ccTGA, ventricular septal defect, and pulmonary stenosis using left ventricular unloading techniques via a bidirectional shunt. PMID- 12740770 TI - Arrhythmia surgery in association with complex congenital heart repairs excluding patients with fontan conversion. AB - Surgical arrhythmia therapy may be performed for patients failing the catheter ablation approach or incorporated into repair of complex congenital heart disease. Variations in atrial and ventricular anatomy that may limit the catheter approach can be directly addressed surgically assuring lesion depth and continuity of anatomic lines of block. Between July 1992 and August 2002, we performed arrhythmia surgery on 34 patients for refractory atrial (n = 29) or ventricular (n = 5) arrhythmias. Not included in this series are patients who had arrhythmia surgery during Fontan conversion. The majority of patients had various forms of complex congenital heart disease; two had structurally normal hearts. Median age at surgery was 13.0 years (range, 7 days to 45 years). Five patients were infants (mean age, 25 days). Twenty-two patients (65%) had an average of 2.8 previous cardiac procedures; all required resternotomy. Operative mortality was 5.9% (2 of 34 patients) because of low cardiac output in one patient following Mustard takedown and arterial switch operation and in one neonate with Ebstein's anomaly and pulmonary atresia. Ablative surgery for supraventricular tachycardia (atrial re-entry, automatic atrial, atrioventricular nodal re-entry, and atrial fibrillation) had a 93% success rate (25 of 27 patients). Clinical tachycardia recurred in two of 27 surviving patients (7%) with atrial arrhythmia, one after an arrhythmia-free interval of several years. Ventricular tachycardia was inducible postoperatively in two of three patients with ventricular arrhythmias and congenital heart disease. Patient size or anatomic complexity should not be limiting factors in the combined surgical arrhythmia approach. Because older patients undergoing surgical revision of prior surgical repairs of congenital heart disease are at increased risk for the later development of atrial arrhythmias, incorporation of arrhythmia therapy into any planned surgical revision should be routinely considered. PMID- 12740771 TI - Electrophysiologic surgery in patients with congenital heart disease. AB - As patients with congenital heart disease increase in number and age, arrhythmia is becoming a more prevalent and important clinical problem. Although catheter based therapy has revolutionized the management of arrhythmia, there remains an increasing patient population with congenital heart disease presenting for repair or reoperation with associated atrial or ventricular arrhythmias. Arrhythmia ablation may be safely and effectively included as an adjunct to repair of underlying structural cardiac lesions. Successful electrophysiologic surgery requires accurate preoperative characterization of the arrhythmia. PMID- 12740772 TI - Surgical management of atrial tachyarrhythmias associated with congenital cardiac anomalies: Mayo Clinic experience. AB - Patients with congenital cardiac anomalies that cause right or left atrial dilatation may have associated atrial tachyarrhythmias, including atrial fibrillation and atrial flutter. Congenital cardiac anomalies may also be associated with Wolff-Parkinson-White syndrome or atrioventricular nodal re entrant tachycardia. In addition, atrial arrhythmias may develop late after definitive operation for congenital cardiac anomalies, especially after the Fontan procedure. Ebstein's anomaly is the most common congenital cardiac anomaly associated with atrial arrhythmias. Atrial arrhythmias cause significant morbidity and mortality, as well as sudden death. Advances in electrophysiologic catheterization and surgical techniques have allowed the diagnosis, localization, and successful treatment of these arrhythmias. PMID- 12740773 TI - Application of robotics in congenital cardiac surgery. AB - Over the past 5 years, robotic systems that combine advanced endoscopic imaging with computer-enhanced instrument control have been used for both coronary revascularization and intracardiac procedures in adults. In addition, endoscope positioning systems and articulated instruments with a robotic wrist mechanism have further expanded the potential applications for robotics in cardiac surgery. In pediatric cardiac surgery, potential applications can be divided into simple scope manipulation versus the use of 3-dimensional imaging and a robotic wrist for dissection and reconstruction. A voice-controlled robotic arm for scope manipulation can facilitate current pediatric thoracoscopic procedures such as ligation of patent ductus arteriosus and division of vascular rings. By using an advanced imaging system along with a robotic wrist, more complex extracardiac and even intracardiac procedures can be performed in children. Examples include coarctation repair, septal defect repair, and mitral or tricuspid valvuloplasty. Furthermore, with adequate intracardiac imaging, a robot-assisted off-pump approach to intracardiac pathology is conceivable. New real-time 3-dimensional echocardiography now offers sufficient resolution to enable such procedures, while the addition of instrument tracking, haptic feedback, and novel tissue fixation devices can facilitate safe and reliable intracardiac repair without extracorporeal circulation. PMID- 12740774 TI - The role of intraoperative device closure in the management of muscular ventricular septal defects. AB - The contemporary surgical management of multiple ventricular septal defects (VSDs) carries a relatively high mortality rate in comparison to that of other biventricular repairs. Several innovative approaches have been recommended for repair of multiple VSDs, but none are uniformly successful in achieving complete elimination of ventricular shunting without causing ventricular dysfunction. This is especially true in cases with highly deficient ventricular septa, the so called "Swiss cheese" anomaly. The recent introduction of the CardioSEAL (NMT Medical Inc, Boston, MA) double-clamshell device led to an evaluation of intraoperative device-mediated closure of muscular VSDs. Taken in context with our overall experience in the surgical management of multiple VSDs, we suggest that device-mediated closure of muscular VSDs is a useful and potentially ventricular function-sparing option for this challenging problem. PMID- 12740775 TI - The hemi-Fontan operation. AB - In hearts with a functional single ventricle, cavity volume and myocardial muscle mass increase as a consequence of the excessive volume load associated with parallel pulmonary and systemic circulations. The hemi-Fontan operation was conceived as a means of accomplishing early reduction of the volume work of the single ventricle, in anticipation of eventual completion of a modified Fontan procedure. The hemi-Fontan procedure includes association of the superior vena(e) cava(e) (SVC) with the branch pulmonary arteries, augmentation of the central pulmonary arteries, occlusion of the inflow of the SVC into the right atrium, and elimination of other sources of pulmonary blood flow. Because it is preparatory to an eventual completion Fontan procedure, concomitant procedures are conveniently performed in conjunction with the hemi-Fontan. These may include atrial septectomy, relief of aortic arch obstruction, proximal main pulmonary artery to ascending aortic anastomosis, repair or revision of anomalous pulmonary venous connection, atrioventricular valvuloplasty, or other procedures. PMID- 12740776 TI - Left ventricular outflow tract obstruction: the role of the modified Konno procedure. AB - The progressive nature of obstruction in the left ventricular outflow tract (LVOT) is hypothesized to be related to the complex relationship between anatomic substrate, genetic predisposition, local shear stress, and the response to injury. Consequently, recurrent obstruction after resection of subaortic membranes as well as more complex forms of LVOT obstruction (eg, tunnel-like stenosis) require aggressive resection to provide unobstructed egress of blood from the left ventricle. The modified Konno procedure is well suited to provide maximal enlargement of the entire LVOT with preservation of aortic valve function. Extension of the procedure to patients with simple forms of LVOT obstruction is, at present, not warranted because of the inability to identify patients destined to develop recurrent obstruction. As predictive models improve, however, the modified Konno procedure should be evaluated in the setting of simple LVOT obstruction at high risk for recurrent obstruction. PMID- 12740777 TI - Re-operative surgery in pediatric patients. AB - In spite of recent emphasis on primary repair for congenital heart defects, multiple operations are often required for staged repairs of complex lesions or replacement of outgrown or degenerated prosthetic material. The vast majority of re-do operations proceed without incident; however, re-operation entails greater risk of inadvertent injury to the phrenic nerve and other heart and vascular structures, postoperative bleeding, and may require alternative cannulation sites. Recommendations to address these hazards are reviewed, as are areas of research that may impact future approaches to the patient who requires re operation. Pericardial closure and pericardial substitutes, bovine and synthetic, are briefly discussed, as is the use of aprotinin for its hemostatic and anti inflammatory effects. PMID- 12740778 TI - The clinical significance of the reoxygenation injury in pediatric heart surgery. AB - Significant advances have been made in the technical performance of operations for infants and neonates with congenital heart disease. However, postoperative organ dysfunction is a frequent problem, particularly in hypoxic (cyanotic) infants. We review both our experimental and subsequent clinical experience with the injury caused by abrupt reoxygenation of the hypoxic patient and examine the modalities of gradual reoxygenation and leukodepletion in limiting this injury, thereby improving operative outcomes for cyanotic lesions. As a result of our experimental and clinical experience we conclude that: (1). reoxygenation injury is a real source of postoperative cardiac and pulmonary dysfunction; (2). white blood cells play an integral role in the production of oxygen-free radicals that are responsible for the damage; and (3). this injury can be modified and possibly ameliorated by changes in the intraoperative management of cardiopulmonary bypass. PMID- 12740779 TI - The effect of modified ultrafiltration on the postoperative course in patients with congenital heart disease. AB - Improvements in the technology of cardiopulmonary bypass have significantly reduced morbidity following repair of congenital cardiac defects. However, the use of cardiopulmonary bypass exposes infants to extremes of hemodilution and hyperthermia, often in association with tissue ischemia. Exposure of the blood to surfaces of the bypass circuit initiates a systemic inflammatory response that may result in organ dysfunction after cardiopulmonary bypass, especially the heart, lungs, and brain. The technique of modified ultrafiltration (MUF) was introduced by Naik and colleagues at the Hospital for Sick Children in London over 10 years ago. Since that time, multiple studies have evaluated the effects of MUF on organ function and postoperative morbidity following repair of congenital heart defects. Use of MUF after cardiopulmonary bypass reverses hemodilution and decreases tissue edema resulting in improved pulmonary function with decreased duration of postoperative ventilation, improved left ventricular function, decreased postoperative bleeding, and a decrease in the incidence and duration of pleural effusions following the Fontan procedure. Despite the increasing evidence that the use of MUF reduces postoperative morbidity, many important questions remain unresolved. The mechanisms by which MUF results in these beneficial effects requires additional investigation. In addition, further studies are necessary to identify patients most likely to benefit from MUF and to define the optimal protocols for its use. In the future, prospective randomized studies incorporating recent advances in the technology of cardiopulmonary bypass will be necessary to define the optimal utilization of ultrafiltration during and after cardiopulmonary bypass. PMID- 12740780 TI - Optimizing response of the neonate and infant to cardiopulmonary bypass. AB - Despite the ability to surgically correct complex defects in neonates and infants, limitations in outcome are sometimes encountered in relation to exposure to cardiopulmonary bypass (CPB). The deleterious effects of CPB in neonates are often pronounced because of their immature tissue/organ function and the disparity between the CPB circuit size and the patient. A variety of techniques that have been developed to prevent or lessen tissue edema, including miniaturization of the circuit and oxygenator, prime additives (eg, albumin, steroids), biocompatible circuitry, and variations in perfusion strategies, are discussed as are post-CPB strategies such as modified ultrafiltration, which removes inflammatory mediator-rich fluid from the patient and bypass circuit. With optimization of the response of the infant to our systems used during repair of their cardiac lesion, we will see significant improvement in surgical outcomes. PMID- 12740781 TI - Ethical issues concerning cardiac surgery in patients with syndromic abnormalities. AB - Advances in surgical techniques, cardiac anesthesia, and pre- and postoperative care have made the surgical treatment of complex congenital cardiac disease available to an ever-increasing number of children, including those with a wide range of extracardiac anomalies. Over the past few decades cardiac surgery in infants and children with syndrome-associated physical and mental conditions has undergone a remarkable change, with previously held norms abandoned for new standards. The social, ethical, and clinical appropriateness of these changes has been the focus of much attention. In this article, we provide a brief history of cardiac surgery in children with congenital syndromes, discuss some groundbreaking cases such as that of "Baby Doe," and present some rules of thumb for the pediatric cardiac surgeon and cardiologist to use when caring for children with congenital syndromes. PMID- 12740782 TI - Ethical and regulatory issues concerning engineered tissues for congenital heart repair. AB - Recent progress in the fields of tissue engineering and xenotransplantation has brought the reality of using engineered tissues for the treatment of congenital heart disease ever closer. However, the introduction of complex scientific advances into the clinic can generate difficult ethical dilemmas for surgeons, patients, and the wider public. Conventional regulatory approaches are not well suited to the introduction of novel cell- and tissue-based therapies. This review presents a short summary of the current state of the art of tissue engineering and xenotransplantation as it relates to congenital heart surgery. The ethical arguments and emerging regulatory framework are then presented, with emphasis on the regulation of tissue-engineered heart valves and the ethics of cardiac xenotransplantation. PMID- 12740783 TI - Have we met the expectations of heart and lung transplantation in children? AB - Heart transplantation in children has been a relatively common practice for more than 15 years, and lung transplantation, while in use as a therapeutic modality for a shorter period of time, has 10 years of follow-up data for review. Because they are expensive, time-consuming, labor-intensive procedures, it is reasonable to review what has transpired to evaluate the effectiveness of these procedures, not only from the standpoint of survival, but in the applicability and availability to all who need it, and successful long-term outcomes including quality of life in those who receive it. Health care expense can be justified by improved results that reach expectations in the areas of applicability, availability, and survival. The applicability issue has, in large part, been achieved in transplantation in children. The goal of comprehensive availability can be met by providing alternatives to transplantation, advancing artificial organ research, and overhauling the organ donor programs to improve organ retrieval. To better meet expectations for survival, further advances in transplant immunology and solutions to the problems of post-transplant coronary artery disease and bronchiolitis obliterans will have to occur. PMID- 12740784 TI - Performance measurement in congenital heart surgery: benefits and drawbacks. AB - Performance monitoring is playing an increasing role in modern congenital heart surgery. We present a number of examples to show the kinds of benefits that it can lead to. Conversely, we discuss some of the potential pitfalls including: errors in the collection of data; use of inappropriate data originally collected for a different purpose; assessment based on inadequate sample size; lack of confidence intervals in reported results; failure to include all members of the team in the assessment; and others. We emphasize that apparent changes in performance from year to year or between different units can be purely due to chance. However, despite these difficulties, we believe that performance monitoring, if carried out carefully, with clearly specified goals, can have many benefits in improving patient care. We suggest that statistical analysis of results should not be taken as a definitive indication of inadequate performance; confidence intervals for any performance measurements should always be clearly indicated; small groups of patients/operations should not be used as a basis for assessment; and that performance measurements should be assigned to the whole team, rather than individual members. PMID- 12740785 TI - Facts and fantasies of congenital heart surgery: thoughts on advances in heart surgery and those who make them. AB - For many years physicians, and surgeons in particular, have been able to implement new treatments with minimal oversight. However, the potential for abuse inherent in such a system has led to an increased degree of monitoring and new regulations. Although regulatory control is far different today, it must be acknowledged that surgeons have enjoyed a greater degree of freedom than other specialists. The congenital heart surgeon's laboratory is, of necessity, often the operating room itself because appropriate animal models do not exist and have not generally been exploited. Advances will surely continue to be made, but the days of innovation as we have come to know them may be lost. In this article, some thoughts regarding advances in cardiac surgery and how surgeons have made them along with insights into the mind and motivation of the heart surgeon are discussed. PMID- 12740786 TI - Impact of donor infections on outcome of orthotopic liver transplantation. AB - Infection occurs when microbial agents enter the host, either through airborne transmission or by direct contact of a substance carrying the infectious agent with the host. Human body fluids, solid organs, or other tissues often are ideal vectors to support microbial agents and can transmit infections efficiently from donor to recipient. In the case of blood transfusion and tissue transplantation, the main consequence of such a transmission is infection of the recipient. However, in the case of solid-organ transplantation, and particularly for liver transplantation, donor infections are not only transmitted to the recipient, the donor infection also may affect the donated liver's preservability and subsequent function in the recipient irrespective of the systemic consequences of the infection. In addition, solid organ recipients of infected organs are less able to respond to the infectious agent because of their immunosuppressive treatment. Thus, transmission of infections from organ donor to liver recipient represents serious potential risks that must be weighed against a candidate's mortality risk without the transplant. However, the ever-increasing gap between the number of donors and those waiting for liver grafts makes consideration of every potential donor, regardless of the infection status, essential to minimize waiting list mortality. In this review, we will focus on assessing the risk of transmission of bacterial, fungal, viral, and parasitic infectious agents from cadaveric liver donors to recipients and the effect such a transmission has on liver function, morbidity, and mortality. We will also discuss risk-benefit deliberations for using organs from infected donors for certain types of recipients. These issues are critically important to maximize the use of donated organs but also minimize recipient morbidity and graft dysfunction. PMID- 12740787 TI - Sirolimus as primary immunosuppression in liver transplantation is not associated with hepatic artery or wound complications. AB - Sirolimus is a new immunosuppressive agent increasingly being used in liver transplant recipients. There is concern that sirolimus may be associated with wound complications and hepatic artery thrombosis (HAT). We have used sirolimus as primary immunosuppression in 170 liver transplant recipients and therefore reviewed our experience with wound complications and HAT in our cohort of patients. Records of all 170 patients administered sirolimus as primary immunosuppression and 180 historic controls were reviewed. Numbers of wound and hepatic artery complications were recorded, as well as the prevalence of obesity, reoperation, diabetes, and OKT3 use, all of which are risk factors for wound complications. The prevalence of wound complications was 12.4% in sirolimus treated patients compared with 13.9% in historic controls (P = not significant [NS]). The prevalence of hepatic artery complications was 5.3% in sirolimus treated patients compared with 8.3% in historic controls (P = NS). The prevalence of obesity and OKT3 administration was significantly lower in sirolimus-treated patients. Multivariate analysis failed to show an association between sirolimus therapy and hepatic artery or wound complications. The prevalence of wound and hepatic artery complications is not different in liver transplant recipients administered sirolimus as part of a primary immunosuppressive regimen compared with historic controls. PMID- 12740788 TI - Rapamycin: friend, foe, or misunderstood? PMID- 12740789 TI - MELD score predicts 1-year patient survival post-orthotopic liver transplantation. AB - The Model for End-Stage Liver Disease (MELD) is an important predictor in patients awaiting orthotopic liver transplantation (OLT). However, the model's association with posttransplant patient survival is unclear. We studied 1-year patient survival in 404 adult patients who underwent OLT at the University of California Los Angeles. The hazard rates of patient survival according to the MELD strata and United Network for Organ Sharing (UNOS) statuses were assessed by Proportional Hazard Cox regression analysis. The difference in survival for MELD strata and UNOS status were compared using the Cox model. There was a significant difference in 1-year patient (P =.0006) survival using different MELD strata, whereas there was a trend according to UNOS status (P =.051). Increased rate of death was observed in recipients of OLT with higher MELD scores (> 36, hazard ratio 3.9; 95% CI 1.55, 10.27) and more urgent UNOS status (2A; hazard ratio, 1.99; 95% CI 1.07, 3.7). The MELD stratum is better associated with 1-year patient survival in liver transplant recipients than UNOS statuses. Patient survival was worse with higher MELD scores. PMID- 12740790 TI - A retrospective review of sirolimus (Rapamune) therapy in orthotopic liver transplant recipients diagnosed with chronic rejection. AB - Treatment options are limited for orthotopic liver transplant (OLT) recipients suffering from chronic rejection (CR). We performed a retrospective review of OLT recipients diagnosed with CR and treated with sirolimus. The medical records of all OLT recipients treated with sirolimus between October, 1998 and October, 2000 were retrospectively reviewed. The diagnosis of CR was made by both clinical and histologic criteria: bile duct to hepatic artery ratio less than 0.7, histologic activity index, hepatic arterial wall thickening, and chronic elevation of liver chemistries. Two groups were defined in regard to sirolimus response: sirolimus responders (SR) and sirolimus nonresponders (SNR). Response to treatment was granted only when patients were found to have resolution of abnormal liver transaminases and an improvement in hepatic artery to bile duct ratio. Serum collections for liver chemistries were collected on days 1, 30, 60, and 90. Liver biopsies were reviewed in blinded fashion from day 1 and at least 180 days on therapy by double-blinded pathologists. Sirolimus-related complications were recorded and include drug toxicity, anemia with and without treatment, hospitalizations, infections, immunosuppression complications, lipid profile disorders, edema, muscle aches, and gastrointestinal complaints. Twenty-one patients were diagnosed with CR. The SR group included 13 of 21, and 8 of 21 were in the SNR group. Anemia was diagnosed in 12 of 21 patients: SR, 7 of 13; SNR, 5 of 8; with 5 patients requiring red blood cell transfusions (2 SR, 3 SNR). Recombinant erythropoietin was started in 5 of 21 patients. Sirolimus serum levels were found to be greater than 20 ng/dL in 12 patients. Sirolimus was discontinued in 9 patients, PMID- 12740791 TI - Effect of chronic administration of tacrolimus and cyclosporine on human gastrointestinal permeability. AB - The antirejection drug tacrolimus (FK506) has been reported to impair intestinal permeability in an early stage after orthotopic liver transplantation (OLT), and cyclosporine (CsA) has shown a similar effect in animals. We studied the chronic effect of FK506 and CsA on gastroduodenal and intestinal permeability and on blood endotoxin levels in patients 2 to 3 years after OLT. Thirty-two OLT patients (22 men and 10 women; mean age, 44.8 +/- 7.1) who had received CsA (n = 19) or FK506 (n = 13) and 10 healthy volunteers (6 male and 4 female, mean age 41.7 +/- 5.4) were assessed for gastroduodenal permeability by recovery in urine of sucrose after oral administration and for intestinal permeability by recovery in urine after oral loads of rhamnose and lactulose, which evaluate the intracellular and paracellular routes, respectively. In all subjects, plasma levels of endotoxins also were assessed. Gastroduodenal permeability was similar in patients and controls (0.03 +/- 0.003 versus 0.04 +/- 0.01%, P = NS). In regard to intestinal permeability, passage through the intracellular route was significantly reduced in OLT patients compared with controls (1.13 +/- 0.06 versus 2.74 +/- 0.17%, P <.01), but paracellular permeability was unchanged (0.14 +/- 0.007 versus 0.13 +/- 0.01%, P = NS). Serum endotoxin levels were similar in all subjects. We conclude that chronic administration of FK506 or CsA induces a clinically irrelevant, selective dysfunction of monosaccharide absorption, but does not affect gastroduodenal or intestinal permeability. PMID- 12740792 TI - Hepatic allograft-derived Kupffer cells regulate T cell response in rats. AB - In liver transplantation, the development of tolerance is associated with an increased rate of apoptosis of T lymphocytes in the portal inflammatory infiltrate and the presence of an intragraft Th2-like T cell population. Underlying mechanisms are poorly understood. Kupffer cells (KC), which reside in the hepatic sinosoids, can directly interact with circulating T lymphocytes and thus are uniquely positioned to play a role in immunomodulation. In this study, the immunoregulatory effects of KC were investigated. We show that KC can significantly suppress T cell proliferation in mixed leukocyte reaction (MLR). Furthermore, KC express functional Fas ligand (FasL) and can induce apoptosis of Fas+ cells. This process can be blocked by addition of neutralizing anti-FasL antibody. Moreover, using an allogeneic liver transplant model we have determined that 1. KC recovered from chronically accepted hepatic allografts have increased FasL messenger RNA (mRNA) and protein expression and a greater ability to induce apoptosis of alloreactive T cells compared with KC recovered from an acute rejection model; 2. KC not only induce apoptosis of T cells, but also regulate cytokine production and Th2/Th3-like cytokine (interleukin [IL]-10 / transforming growth factor [TGF]-beta) mRNA expression in allogeneic MLR in vitro; and 3. administration of KC derived from chronically accepted liver allografts significantly prolongs the survival of hepatic allografts in an acute rejection model in an alloantigen-specific manner. In conclusion, these data implicate the possible role of KC-mediated regulation of T cell response in the induction of immune tolerance in liver allografts. PMID- 12740793 TI - Kupffer cells: another player in liver tolerance induction. PMID- 12740794 TI - Clinical experience gained from the use of 120 steatotic donor livers for orthotopic liver transplantation. AB - Steatosis of the donor liver is known to impact on patient and allograft outcome after orthotopic liver transplantation (OLT). The aim of this study is to evaluate the effect of increasing grades of cadaveric donor liver steatosis on recipient outcome. Between January, 1986 and December, 2000, 120 OLTs were performed with 72 mild, 25 moderate, and 23 severe steatotic donor livers. Donors of steatotic livers were more likely to be older (P =.001) and have died of intracerebral haemorrhage than donors of nonsteatotic livers. Initial poor graft function (IPF) was more common in donor livers with either moderate or severe steatosis than in donor livers with mild steatosis (P =.03). Primary graft nonfunction (PNF) occurred in only 1 donor liver with severe steatosis. PGE1 (PGE1) usage was higher in recipients of donor livers with moderate or severe steatosis versus donor livers with mild steatosis (P =.001). Allograft loss was greater at 1 year both in the moderate and severe (P =.03) steatotic liver groups. Patient survival at 3 months and overall allograft survival both were impacted negatively by increasing grades of donor liver steatosis (P =.02, P =.03). Three-month allograft survival was reduced in the steatotic donor livers if the donor was 50+ years old (P =.033). Recipient status at OLT (P =.001) and donor steatosis (P =.046) impacted on 30 day allograft survival (multivariate analysis). In conclusion, increasing grades of donor liver steatosis were associated with worse IPF and increased PGE1 usage. There was a negative impact of steatosis on both recipient and early allograft survival. PMID- 12740795 TI - Isolation of human hepatocytes from livers rejected for liver transplantation on a national basis: results of a 2-year experience. AB - The offer of liver transplantation to many patients affected by liver failure is limited by organ shortage. Clinical application of human-based liver cell therapies, such as bioartificial liver and hepatocyte transplantation, might support liver transplantation, allowing more patients to be treated and decreasing mortality in the waiting list. The development of a standardized method of hepatocyte isolation is a mainstay for large-scale application of liver cell therapy. The aim of this study is to analyze retrospectively a 2-year experience of human hepatocyte isolation from livers rejected from transplantation at organ harvesting, performed on a national basis in Italy. All the livers judged unsuitable for transplantation were considered for hepatocyte isolation. Macrosteatosis greater than 60% was the most common reason of refusal, followed by nonviral cirrhosis. Fifty-four organs were used. Human hepatocyte isolation resulted in more that 7 million liver cells/g of tissue digested with 73% +/- 14% viability. Steatotic organs gave better results in terms of cell yield than cirrhotic livers. Isolated hepatocytes were able to perform specific liver functions, and evidence of factor IX and albumin messenger RNA (mRNA) production was reported when cells were plated in culture. Modifications of the traditional method of hepatocyte isolation, aimed at reducing ischemia reperfusion damage and improving post-isolation cell conditions, showed improvements in post-isolation viability. In conclusion, we show that it is possible to use the vast majority of livers not suitable for transplantation on a national basis for human hepatocyte isolation, obtaining a large amount of viable functioning human hepatocytes that might be used for cell transplantation and therapy. PMID- 12740797 TI - Is liver transplantation justified for the treatment of HCC in Child's A patients? Not always. PMID- 12740796 TI - Small hepatocellular carcinoma in Child A cirrhotic patients: hepatic resection versus transplantation. AB - Hepatic resection (HR) is the treatment of choice for small hepatocellular carcinoma (HCC) in a noncirrhotic liver, whereas liver transplantation (LT) offers better results in patients with impaired hepatic function (Child B and C). However, it is still debated whether HR or LT is the best strategy for patients with Child A cirrhosis. We conducted a retrospective study on 37 consecutive patients with Child A cirrhosis and small HCC, treated between 1991 and 1999. Seventeen of these patients, who underwent LT, were compared with 20 patients who underwent HR, and prognostic factors for survival and tumor recurrence were analyzed. The primary endpoints were the intention-to-treat, 3- and 5-year survival, and 3- and 5-year recurrence-free survival. Three- and 5-year patient survival rate both were significantly (P =.04) higher in the LT group (87% and 71%, respectively) than in the HR group (67 and 36% respectively). Similarly, the 3- and 5- year recurrence-free survival rates were 87% and 80% for the LT group, and 52% and 40% for the HR group (P =.03). Absence of microscopic vascular invasion was the only other prognostic factor correlated with significantly better recurrence-free survival (P =.02). Therefore, we concluded that in patients with Child A cirrhosis and small HCC, liver transplantation resulted in better overall and disease-free survival than HR. PMID- 12740798 TI - Acquired hemophilia A after liver transplantation: a case report. AB - It is well recognized that orthotopic liver transplantation rapidly corrects the coagulation disorder observed in patients with hemophilia A or B combined with end-stage liver disease; however, the transmission of hemophilia from a donor to a transplant recipient has not been reported previously. We describe a patient who developed acquired hemophilia A after liver transplantation due to the unknown presence of factor VIII inhibitor in the donor. The resulting coagulation disorder was unresponsive to standard factor replacement therapy, and was ultimately corrected with retransplantation. Hemophiliac donors should not be precluded from organ donation, though the presence of factor VIII inhibitor should be actively sought before organ procurement. High levels of factor VIII inhibitor represents an absolute contraindication to liver donation. PMID- 12740799 TI - A model to predict survival at one month, one year, and five years after liver transplantation based on pretransplant clinical characteristics. AB - Reliable models that could predict outcome of liver transplantation (LT) may guide physicians to advise their patients of immediate and late survival chances and may help them to optimize organ use. The objective of this study was to develop user-friendly models to predict short and long-term mortality after LT in adults based on pre-LT recipient characteristics. The United Network for Organ Sharing (UNOS) transplant registry (n = 38,876) from 1987 to 2001 was used to develop and validate the model. Two thirds of patients were randomized to develop the model (the modeling group), and the remaining third was randomized to cross validate (the cross-validation group) it. Three separate models, using multivariate logistic regression analysis, were created and validated to predict survival at 1 month, 1 year, and 5 years. Using the total severity scores of patients in the modeling group, a predictive model then was created, and the predicted probability of death as a function of total score then was compared in the cross-validation group. The independent variables that were found to be very significant for 1 month and 1 year survival were age, body mass index (BMI), UNOS status 1, etiology, serum bilirubin (for 1 month and 1 year only), creatinine, and race (only for 5 years). The actual deaths in the cross-validation group followed very closely the predicted survival graph. The chi-squared goodness-of fit test confirmed that the model could predict mortality reliably at 1 month, 1 year, and 5 years. We have developed and validated user-friendly models that could reliably predict short-term and long-term survival after LT. PMID- 12740800 TI - To transplant, or not to transplant? PMID- 12740801 TI - The impact of advancing donor age on histologic recurrence of hepatitis C infection: the perils of ignored maternal advice. PMID- 12740802 TI - Environmental exposure to mercury and its toxicopathologic implications for public health. AB - Mercury is a toxic and hazardous metal that occurs naturally in the earth's crust. Natural phenomena such as erosion and volcanic eruptions, and anthropogenic activities like metal smelting and industrial production and use may lead to substantial contamination of the environment with mercury. Through consumption of mercury in food, the populations of many areas, particularly in the developing world, have been confronted with catastrophic outbreaks of mercury induced diseases and mortality. Countries such as Japan, Iraq, Ghana, the Seychelles, and the Faroe Islands have faced such epidemics, which have unraveled the insidious and debilitating nature of mercury poisoning. Its creeping neurotoxicity is highly devastating, particularly in the central and peripheral nervous systems of children. Central nervous system defects and erethism as well as arrythmias, cardiomyopathies, and kidney damage have been associated with mercury exposure. Necrotizing bronchitis and pneumonitis arising from inhalation of mercury vapor can result in respiratory failure. Mercury is also considered a potent immunostimulant and -suppressant, depending on exposure dose and individual susceptibility, producing a number of pathologic sequelae including lymphoproliferation, hypergammaglobulinemia, and total systemic hyper- and hyporeactivities. In this review we discuss the sources of mercury and the potential for human exposure; its biogeochemical cycling in the environment; its systemic, immunotoxic, genotoxic/carcinogenic, and teratogenic health effects; and the dietary influences on its toxicity; as well as the important considerations in risk assessment and management of mercury poisoning. PMID- 12740803 TI - Toxicological comparison of diverse Cylindrospermopsis raciborskii strains: evidence of liver damage caused by a French C raciborskii strain. AB - The freshwater cyanobacterium Cylindrospermopsis raciborskii is known to produce toxic effects in several countries. Acute and chronic exposures to C. raciborskii in Australia have been linked to liver damage (hepatotoxicity) with concomitant effects on the kidneys, adrenal glands, small intestine, lungs, thymus, and heart. The alkaloid cylindrospermopsin, which produces these toxic effects, is thought to be a potent inhibitor of protein synthesis. C. raciborskii strains producing cylindrospermopsin or analogue alkaloids have also been reported in Florida, USA, and Thailand. Brazilian isolates of C. raciborskii are also toxic but act by a different mechanism, causing acute death in mice with neurotoxic symptoms similar to those induced by the saxitoxins. In this article we compare the toxicity in the mouse of a C. raciborskii French strain with C. raciborskii strains from various other sources (Australia, Brazil, Mexico, and Hungary). We tested the toxicity of cell extracts by a mouse bioassay. Acute, fatal neurotoxicity was produced by the Brazilian strain, which was confirmed by liquid chromatography with fluorescence detection of the cell extracts, which revealed the presence of saxitoxin, neosaxitoxin, and decarbamoylsaxitoxin, along with two unidentified compounds. Acute hepatotoxicity with severe liver, kidney, and thymus damage was observed with the Australian cylindrospermopsin-producing strain. The Mexican and Hungarian strains were not found to be toxic to mice in our experimental conditions. No animals died after exposure to the extracts of the French C. raciborskii strain. Histological examination of the liver revealed moderate, multifocal necrosis characterized by small areas of hepatocellular necrosis, combined with disorganization of the parenchyma and congestion of the inner sinusoid. These symptoms and lesions resembled those induced by cylindrospermopsin, but the chemical analysis performed by liquid chromatography coupled with either a diode array detector or a mass spectrometer demonstrated that this toxin was not present in our culture extract. PMID- 12740804 TI - Microbiological evaluation of toxicity of three polycyclic aromatic hydrocarbons and their decomposition products formed by advanced oxidation processes. AB - The toxicity of benzo[a]pyrene, chrysene, and fluorene and their decomposition products formed by advanced oxidation processes (AOPs) was investigated using biotests with Escherichia coli and Vibrio fischeri. The polycyclic aromatic hydrocarbons (PAHs) were not highly toxic to either bacterial strain; the toxicity of their degradation products depended on the method of chemical processing. Inhibition of more than 27% was observed with products formed by oxidation of the PAHs, by AOP methods without hydrogen peroxide. Toxicity as high as 100% was observed after the combined action of hydrogen peroxide and other oxidizing agents. PMID- 12740805 TI - Occurrence of toxic cyanobacterial blooms in San Roque Reservoir (Cordoba, Argentina): a field and chemometric study. AB - We evaluated the presence of cyanobacterial blooms in San Roque Reservoir (Cordoba, Argentina). Cyanobacterial blooms and water samples were collected over 4 years (1998-2002). We confirmed the presence of microcystin-LR and microcystin RR in 97% of these blooms. The total amount of microcystin (MC) ranged between 5.8 and 2400.0 microg x g(-1) of freeze-dried bloom material. These values suggest that guidelines for safe water consumption and recreational use should be established for this reservoir. Twenty-eight physical and chemical parameters were measured in water samples and evaluated by discriminant analysis (DA). A first DA was used to evaluate the factors promoting cyanobacteria occurrence, identifying nine parameters following three patterns associated with cyanobacterial growth. Inorganic phosphorous was found to promote the presence of blooms, whereas the highest proliferation of cyanobacteria was observed in the presence of smaller amounts of carbonate, bicarbonate, sulfate, and fecal coliform bacteria. The results observed during our fieldwork, analyzed using DA, agreed with the results of other laboratory studies, thus confirming the usefulness of DA to help with the evaluation of a complicated environmental data matrix. A second DA, using only water samples collected during the presence of cyanobacteria blooms, identified another nine parameters. The analysis of these parameters allowed us to identify certain environmental factors that could lead to the dominance of toxic strains, thus increasing the amount of MC. The results showed that, in our case, an increase in the water temperature was associated with higher amounts of MC per dry weight unit, whereas an increase in the concentrations of ammonia-nitrogen and iron were associated with lower amounts of MC, thus disfavoring the dominance of toxic strains. PMID- 12740806 TI - Effects of microcystin-free and microcystin-containing strains of the cyanobacterium Microcystis aeruginosa on growth of the grazer Daphnia magna. AB - Harmful effects of the common bloom-forming cyanobacterium Microcystis aeruginosa on the grazer Daphnia have been explained from morphological features, nutritional insufficiency, and the production of toxins called microcystins. The effects of four M. aeruginosa strains, including one free of microcystins, on the growth of Daphnia magna were examined. Population growth was measured in Daphnia fed each M. aeruginosa strain alone and fed mixtures of each strain with the green alga Scenedesmus obliquus to avoid nutritional deficiency. The presence of Microcystis in the food resulted in decreased Daphnia population growth rates. A significant correlation (P < 0.001) was found between the amount of microcystins (MC) in the food and the population growth rate, but growth inhibition in Daphnia fed with the MC-free strain could not be explained by the toxicity of the microcystins. Inasmuch as there was greater reduction in growth of the Daphnia fed mixtures of the MC-free strain with Scenedesmus than of the Daphnia fed only the same amount of Scenedesmus as in the mixtures, other factors associated with this Microcystis strain must have inhibited growth. However, morphology and nutritional insufficiency could be excluded as causal factors. The strong relationship between Daphnia feeding activity and population growth suggests that feeding inhibition may have caused the observed detrimental effects on the animals. PMID- 12740808 TI - Aspartic proteases involved in Alzheimer's disease. AB - Alzheimer's disease afflicts every tenth human aged over 65. Despite the dramatic progress that has been made in understanding the disease, the exact cause of Alzheimer's disease is still unknown. Most gene mutations associated with Alzheimer's disease point at the same culprits: amyloid precursor protein and ultimately amyloid beta. The enigmatic proteases alpha-,beta-, and gamma secretase are the three executioners of amyloid precursor protein processing, and disruption of their delicate balance is suspected to result in Alzheimer's disease. Significant progress has been made in the selective control of these proteases, regardless of the availability of structural information. Not even the absence of a robust cell-free assay for gamma-secretase could hamper the identification of nonpeptidic inhibitors of this enzyme for long. Within five years, four distinctly different structural moieties were developed and the first drug candidates are in clinical trials. Unfortunately, selective inhibition of amyloid beta formation remains a crucial issue because fundamental fragments of the gamma-secretase complex are important for other signaling events. This problem makes beta-secretase inhibition and alpha-secretase induction even more appealing. PMID- 12740809 TI - Signaling effects of demethylasterriquinone B1, a selective insulin receptor modulator. AB - A possible breakthrough in the treatment of diabetes was made with the discovery that a fungal natural product, demethylasterriquinone B1 (DAQ B1), is an orally active, small-molecule mimic of insulin. Subsequent work has shown that the glucose-lowering effects of DAQ B1 are not accompanied by enhanced vascular proliferation, which is a side effect of chronic insulin administration that can lead to arteriosclerosis. Our recent short and modular total synthesis of DAQ B1 could be readily modified to create congeners and afforded ample supplies of the natural product, which permitted intracellular signal transduction of DAQ B1 to be examined. The activities of DAQ B1 and over a dozen related structures were studied for insulin receptor (IR) and insulin receptor substrate-1 phosphorylation. Examination of the effect of DAQ B1 on kinases downstream of the IR in insulin signal transduction showed selective activation of Akt kinase (a metabolic effect) but not of extracellular-regulated kinase (a proliferative effect). The influence of DAQ B1 on gene expression (determined by a microarray study) was also divergent from that of insulin, which activates both proliferative and metabolic pathways. The action of DAQ B1 as a selective insulin receptor modulator can be accounted for by its ability to selectively activate one kinase among the many emanating from insulin receptor autophosphorylation and its reduced effects on gene expression. PMID- 12740810 TI - Indolocarbazole glycosides in inactive conformations. AB - Indolocarbazole glycosides related to rebeccamycin represent a promising category of antitumor agents targeting DNA and topoisomerase I. These drugs prefer to adopt a closed conformation with an intramolecular hydrogen bond between the indole NH group and the pyranose oxygen atom. Three pairs of indolocarbazole monoglycosides bearing an NH or an N-methyl indole moiety were synthesized and their biological properties investigated at the molecular and cellular level. Replacing the indole NH proton with a methyl group reduces DNA interaction and abolishes activity against DNA topoisomerase I. Surface plasmon resonance studies performed with a pair of water-soluble indolocarbazole glycosides and two hairpin oligonucleotides containing an [AT]4 or a [CG]4 sequence indicate that both the NH and the N-methyl derivative maintain a relatively high affinity for DNA (Keq = 2 - 6 x 10(5) M(-1)) but the incorporation of the methyl group restricts access to the DNA. The number of ligand binding sites (n) on the oligonucleotides is about twice as high for the NH compound compared to its N-methyl analogue. Modeling and 1H NMR studies demonstrate that addition of the N-methyl group drives a radical change in conformation in which the orientation of the aglycone relative to the beta-glucoside is reversed. The loss of the closed conformation by the N-methyl derivatives perturbs thir ability to access DNA binding sites and prevents the drug from inhibiting topoisomerase I. As a consequence, the NH compounds exhibit potent cytotoxicity against CEM leukemia cells with an IC50 value in the 1 microM range, whereas the N-methyl analogues are 10 to 100 times less cytotoxic. These studies offer circumstantial evidence supporting the importance of the closed conformation in the interaction of indolocarbazole glycosides with their molecular targets, DNA and topoisomerase I. PMID- 12740811 TI - Monitoring the cellular surface display of recombinant proteins by cysteine labeling and flow cytometry. AB - A general method is described that allows one to follow the surface display of recombinant proteins in Escherichia coli without having to use specific antibodies or enzymatic reactions. The method is based on cysteine-specific labeling through Michael addition to the double bond of maleimide and its derivatives, and takes advantage of the fact that naturally occurring surface proteins in E. coli contain no accessible cysteine residues. The method is easy to perform and could be simply applied to different analytic procedures including Western blot, spectral photometry, and flow cytometry. By using this new labeling method, single cells bearing a distinct protein at the surface could be selected by fluorescence-activated cell sorting. The data were obtained by using autodisplay, an efficient surface display system established for E. coli, but the method presented here represents rather a general solution for analyzing the surface display of recombinant proteins, independent of the cellular system used. PMID- 12740813 TI - Comparison of the chemical properties of iron and cobalt porphyrins and corrins. AB - Density functional calculations have been used to compare various geometric, electronic and functional properties of iron and cobalt porphyrin (Por) and corrin (Cor) species. The investigation is focussed on octahedral M(II/III) complexes (where M is the metal) with two axial imidazole ligands (as a model of b and c type cytochromes) or with one imidazole and one methyl ligand (as a model of methylcobalamin). However, we have also studied some five-coordinate M(II) complexes with an imidazole ligand and four-coordinate M(I/II) complexes without any axial ligands as models of other intermediates in the reaction cycle of coenzyme B12. The central cavity of the corrin ring is smaller than that of porphine. We show that the cavity of corrin is close to ideal for low-spin Co(III), Co(II), and Co(I) with the axial ligands encountered in biology, whereas the cavity in porphine is better suited for intermediate-spin states. Therefore, the low-spin state of Co is strongly favoured in complexes with corrins, whereas there is a small energy difference between the various spin states in iron porphyrin species. There are no clear differences for the reduction potentials of the octahedral complexes, but [Co(I)Cor] is more easily formed (by at least 40 kJ mole(-1)) than [Fe(I)Por]. Cobalt and corrin form a strong Cobond;C bond that is more stable against hydrolysis than iron and porphine. Finally, Fe(II/III) gives a much lower reorganisation energy than Co(II/III); this is owing to the occupied d(z2) orbital in Co(II). Altogether, these results give some clues about how nature has chosen the tetrapyrrole rings and their central metal ion. PMID- 12740812 TI - Large-scale in vivo synthesis of the carbohydrate moieties of gangliosides GM1 and GM2 by metabolically engineered Escherichia coli. AB - Two metabolically engineered Escherichia coli strains have been constructed to produce the carbohydrate moieties of gangliosides GM2 (GalNAcbeta-4(NeuAcalpha 3)Galbeta-4Glc; Gal = galactose, Glc = glucose, Ac = acetyl) and GM1 (Galbeta 3GalNAcbeta-4(NeuAcalpha-3)Galbeta-4Glc. The GM2 oligosaccharide-producing strain TA02 was devoid of both beta-galactosidase and sialic acid aldolase activities and overexpressed the genes for CMP-NeuAc synthase (CMP = cytidine monophosphate), alpha-2,3-sialyltransferase, UDP-GlcNAc (UDP = uridine diphosphate) C4 epimerase, and beta-1,4-GalNAc transferase. When this strain was cultivated on glycerol, exogenously added lactose and sialic acid were shown to be actively internalized into the cytoplasm and converted into GM2 oligosaccharide. The in vivo synthesis of GM1 oligosaccharide was achieved by taking a similar approach but using strain TA05, which additionally overexpressed the gene for beta-1,3-galactosyltransferase. In high-cell-density cultures, the production yields for the GM2 and GM1 oligosaccharides were 1.25 g L(-1) and 0.89 g L(-1), respectively. PMID- 12740814 TI - Semisynthesis and characterization of the first analogues of pro-neuropeptide y. AB - Enzymatic cleavage of prohormone neuropeptide Y (proNPY) leads to mature neuropeptide Y (NPY), a widely distributed neuropeptide with multiple functions both peripherally and centrally. A single dibasic pair of amino acids, Lys38 Arg39, represents the recognition motif for a class of hormone-processing enzymes known as prohormone convertases (PCs). Two members of this PC family, PC1/3 and PC2, are involved in proNPY cleavage. The aim of this work was to establish an effective method for the generation of full-length 69-amino acid proNPY analogues for further studies of prohormone convertase interaction. We have chosen two ligation sites in order to perform the semisynthesis of proNPY analogues by expressed protein ligation (EPL). By using the intein-mediated purification system (IMPACT) with improved conditions for intein splicing, we were able to isolate proNPY 1-40 and proNPY 1-54 fragments as Cterminal thioesters. Peptides bearing Nterminal cysteine instead of the naturally occurring Ser41 and Thr55 residues, respectively, were generated by solid-phase peptide synthesis. Moreover, labels (carboxyfluorescein and biotin) were inserted into the peptide sequences. The synthesis of the [C41]proNPY 41-69 fragment, which proved to be a difficult peptide sequence, could be achieved by the incorporation of two pseudo proline derivatives. Western blot analysis revealed that all five proNPY analogues are recognized by monoclonal antibodies directed against NPY as well as against the Cflanking peptide of NPY (CPON). PMID- 12740815 TI - [7-(Dialkylamino)coumarin-4-yl]methyl-Caged Compounds as Ultrafast and Effective Long-Wavelength Phototriggers of 8-Bromo-Substituted Cyclic Nucleotides. AB - [7-(Dimethylamino)coumarin-4-yl]methyl (DMACM) and [7-(diethylamino)coumarin-4 yl]methyl (DEACM) esters of 8-bromoadenosine 3',5'-cyclic monophosphate (8-Br cAMP) and 8-bromoguanosine 3',5'-cyclic monophosphate (8-Br-cGMP) are described as novel caged compounds for 8-bromo-substituted cyclic nucleotides. Synthesis is accomplished by treatment of the free acids of the cyclic nucleotides with the corresponding 7(dialkylamino)-substituted 4(diazomethyl)coumarins. Irradiation of the DMACM- and DEACM-caged cyclic nucleotides with UV light stimulates the release of the cyclic nucleotides within roughly a nanosecond. The new caged compounds are resistant to hydrolysis in aqueous buffers and exhibit long wavelength absorption properties with maxima at 400 nm, high extinction coefficients, and high quantum yields (0.15-0.31). Their favorable properties render these compounds the most efficient and rapid phototriggers of 8-bromo substituted cyclic nucleotides known. The usefulness of the compounds for physiological studies under nondamaging light conditions was examined in HEK293 cells expressing the alpha subunit of the cyclic-nucleotide-gated (CNG) channel of cone photoreceptors (CNGA3) and of olfactory neurons (CNGA2) by using confocal laser scanning microscopy and the patch clamp technique. PMID- 12740816 TI - Application of the nucleotidylyltransferase Ep toward the chemoenzymatic synthesis of dTDP-desosamine analogues. PMID- 12740819 TI - A review of morphological techniques for detection of peroxisomal (and mitochondrial) proteins and their corresponding mRNAs during ontogenesis in mice: application to the PEX5-knockout mouse with Zellweger syndrome. AB - In the era of application of molecular biological gene-targeting technology for the generation of knockout mouse models to study human genetic diseases, the availability of highly sensitive and reliable methods for the morphological characterization of the specific phenotypes of these mice is of great importance. In the first part of this report, the role of morphological techniques for studying the biology and pathology of peroxisomes is reviewed, and the techniques established in our laboratories for the localization of peroxisomal proteins and corresponding mRNAs in fetal and newborn mice are presented and discussed in the context of the international literature. In the second part, the literature on the ontogenetic development of the peroxisomal compartment in mice, with special emphasis on liver and intestine is reviewed and compared with our own data reported recently. In addition, some recent data on the pathological alterations in the liver of the PEX5(-/-) mouse with a peroxisomal biogenesis defect are briefly discussed. Finally, the methods developed during these studies for the localization of mitochondrial proteins (respiratory chain complexes and MnSOD) are presented and their advantages and pitfalls discussed. With the help of these techniques, it is now possible to identify and distinguish unequivocally peroxisomes from mitochondria, two classes of cell organelles giving by light microscopy a punctate staining pattern in microscopical immunohistochemical preparations of paraffin-embedded mouse tissues. PMID- 12740820 TI - Peroxisome assembly in yeast. AB - Peroxisomes are essential organelles that may be involved in various functions, dependent on organism, cell type, developmental stage of the cell, and the environment. Until recently, peroxisomes were viewed as a class of static organelles that developed by growth and fission from pre-existing organelles. Recent observations have challenged this view by providing evidence that peroxisomes may be part of the endomembrane system and constitute a highly dynamic population of organelles that arises and is removed upon environmental demands. Additionally, evidence is now accumulating that peroxisomes may arise by alternative methods. This review summarizes relevant recent data on this subject. In addition, the progress in the understanding of the principles of the peroxisomal matrix protein import machinery is discussed. PMID- 12740821 TI - Degradation of normal and proliferated peroxisomes in rat hepatocytes: regulation of peroxisomes quantity in cells. AB - Degradation and turnover of peroxisomes is reviewed. First, we describe the historical aspects of peroxisome degradation research and the two major concepts for breakdown of peroxisomes, i.e., autophagy and autolysis. Next, the comprehensive knowledge on autophagy of peroxisomes in mammalian and yeast cells is reviewed. It has been shown that proliferated peroxisomes are degraded by selective autophagy, and studies using yeast cells have been especially helpful in shedding light on the molecular mechanisms of this process. The degradation of extraperoxisomal urate oxidase crystalloid is noted. Overexpressed wild-type urate oxidase in cultured cells has been shown to be degraded through an unknown proteolytic pathway distinct from the lysosomal system including autophagy or the ubiquitin-proteasome system. Finally, peroxisome autolysis mediated by 15 lipoxygenase (15-LOX) is described. 15-LOX is integrated into the peroxisome membrane causing focal membrane disruptions. The content of the peroxisomes is then exposed to cytosol proteases and seems to be digested quickly. In conclusion, the number of peroxisomes appears to be regulated by two selective pathways, autophagy, including macro- and microautophagy, and 15-LOX-mediated autolysis. PMID- 12740822 TI - Selective degradation of peroxisomes in yeasts. AB - In the last two decades, much progress has been made in understanding the process of induction and biogenesis of peroxisomes, essential organelles in all eukaryotes. Only relatively recently, the first molecular studies on the selective degradation of this important organelle-a process known as pexophagy, which occurs when the organelles have become redundant-have been performed, especially using methylotrophic yeasts. The finding that pexophagy and other transport pathways to the vacuole (vacuolar protein sorting, autophagy, cytoplasm to-vacuole-targeting and endocytosis) utilize common but also unique genes has placed pexophagy in the heart of the machinery that recycles cellular material. The quest is now on to understand how peroxisome degradation has become such a highly selective process and what the signals are that trigger it. In addition, because the prime determinant of pexophagy is located on the peroxisome itself, it has become essential to study the role of peroxisomal membrane proteins in the degradation process in detail. This review highlights the main achievements of the last years. PMID- 12740823 TI - Peroxisomal motility and interaction with microtubules. AB - Recent in vivo observations have revealed that peroxisomes are more dynamic and interactive than previously assumed. The growing recognition of the tubular and reticular morphology of peroxisomes in living cells, their association with microtubules, and the dynamic movements of peroxisomes in vivo and in vitro have inspired the query into the investigation of the cellular machinery that mediates such a complex behaviour. The characterisation of the underlying molecular components of this machinery is providing insight into the mechanisms regulating peroxisomal morphology and intracellular distribution. PMID- 12740824 TI - Regulation of cell proliferation and cell death by peroxisome proliferators. AB - Peroxisome proliferators cause increases in liver mass in rodents, linked to changes in cell proliferation and cell death of hepatocytes. These effects are reversible upon cessation of treatment. The underlying mechanism of the response in rodent liver is complex, but clearly dependent on activation of the nuclear receptor PPARalpha. Other signaling pathways have been implicated in this response, but evidence is mixed. Differing sensitivity among various species to effects of peroxisome proliferators has been associated with differences in PPARalpha expression and function. Changes in cell proliferation and cell death in neoplastic hepatocytes also have been found in liver tumors caused by long term treatment with peroxisome proliferators. PMID- 12740825 TI - Induction of the expression of the peroxisome proliferator-activated receptor alpha (PPARalpha) by clofibrate in jerboa tissues. AB - Peroxisome proliferator-activated receptor alpha (PPARalpha) is a member of the nuclear hormone receptor superfamily that can be activated by natural fatty acids and various xenobiotics, including clofibrate. This transcription factor primarily regulates genes involved in lipid metabolism and homeostasis. We present the expression pattern of the PPARalpha subtype in the adult jerboa Jaculus orientalis, determined by RT-PCR and Western blotting using specific probes and a polyclonal antibody for PPARalpha, respectively. PPARalpha is highly expressed in liver and kidney, and to a lesser extent in duodenum and colon. PPARalpha expression is increased at the mRNA and protein levels in liver and duodenum of jerboa treated for 2 weeks with the peroxisome proliferator (PP) clofibrate. The induction is tissue-specific as no significant changes are observed in kidney and colon. The present data indicate that the PP-induced PPARalpha gene expression is not dependent on the PPARalpha content in target cells. PMID- 12740826 TI - Peroxisome proliferation as a biomarker in environmental pollution assessment. AB - Peroxisome proliferators comprise a heterogeneous group of compounds known for their ability to cause massive proliferation of peroxisomes and liver carcinogenesis in rodents. In recent years it has become evident that other animals may be threatened by peroxisome proliferators, in particular aquatic organisms living in coastal and estuarine areas. These animals are exposed to a variety of pollutants of industrial, agricultural and urban origin which are potential peroxisome proliferators. Both laboratory and field studies have shown that phthalate ester plasticizers, PAHs and oil derivatives, PCBs, certain pesticides, bleached kraft pulp and paper mill effluents, alkylphenols and estrogens provoke peroxisome proliferation in different fish or bivalve mollusc species. The response appears to be mediated by peroxisome-proliferator activated receptors, members of the nuclear receptor family, recently cloned in fish. Based on these results it is proposed that peroxisome proliferation could be used as a biomarker of exposure to a variety of pollutants in environmental pollution assessment. This is illustrated by a case study in which mussels, used worldwide as sentinels of environmental pollution, were transplanted from reference to contaminated areas and vice versa. In mussels native to an area polluted with PAHs and PCBs, peroxisomal acyl-CoA oxidase (AOX) activity and peroxisomal volume density were 2-3 fold and 5-fold higher, respectively, compared to the reference site. When animals were transplanted to the polluted station, with increased concentration of organic xenobiotics, a concomitant significant increase of AOX was recorded. Conversely, in animals transplanted to the cleaner station, AOX activity and peroxisomal volume density decreased significantly. These results indicate that peroxisome proliferation is a rapid (i.e., two days) and reversible response to pollution in mussels. Before peroxisome proliferation can be implemented as a biomarker in biomonitoring programs, a well-defined protocol should be established and validated in intercalibration and quality assurance programmes. Furthermore, the influence of biotic and abiotic factors, some of which are known to affect peroxisome proliferation (season, tide level, interpopulation and interindividual variability), should be taken into consideration. The possible hepatocarcinogenic effects as well as the potential adverse effects on reproduction, development, and growth of peroxisome proliferators are unknown in aquatic organisms, thus providing a challenge for future investigations. PMID- 12740827 TI - Human peroxisomal disorders. AB - Peroxisomes are single membrane-bound cell organelles performing numerous metabolic functions. The present article aims to give an overview of our current knowledge about inherited peroxisomal disorders in which these organelles are lacking or one or more of their functions are impaired. They are multiorgan disorders and the nervous system is implicated in most. After a summary of the historical names and categories, each having distinct symptoms and prognosis, microscopic pathology is reviewed in detail. Data from the literature are added to experience in the authors' laboratory with 167 liver biopsy and autopsy samples from peroxisomal patients, and with a smaller number of chorion samples for prenatal diagnosis, adrenal-, kidney-, and brain samples. Various light and electron microscopic methods are used including enzyme- and immunocytochemistry, polarizing microscopy, and morphometry. Together with other laboratory investigations and clinical data, this approach continues to contribute to the diagnosis and further characterization of peroxisomal disorders, and the discovery of novel variants. When liver specimens are examined, three main groups including 9 novel variants (33 patients) are distinguished: (1) absence or (2) presence of peroxisomes, and (3) mosaic distribution of cells with and without peroxisomes (10 patients). Renal microcysts, polarizing trilamellar inclusions, and insoluble lipid in macrophages in liver, adrenal cortex, brain, and in interstitial cells of kidney are also valuable for classification. On a genetic basis, complementation of fibroblasts has classified peroxisome biogenesis disorders into 12 complementation groups. Peroxisome biogenesis genes (PEX), knock-out-mice, and induction of redundant genes are briefly reviewed, including some recent results with 4-phenylbutyrate. Finally, regulation of peroxisome expression during development and in cell cultures, and by physiological factors is discussed. PMID- 12740828 TI - An earlier role for viral pharmacodynamics. PMID- 12740829 TI - Interaction of hepatitis B virus with cells. AB - Virus infection is initiated by recognition and attachment of the virus to the cell surface. Despite the fact that this interaction determines the virus-related pathogenesis, its molecular basis remained obscure for HBV. This process is mediated primarily by the viral envelope and the cellular receptors. HBV infection is not exceptional in this regard but its putative receptors have not been identified yet. The recent development of protocols to establish HBV susceptible cell lines and unique tools to measure HBV-cell attachment at a single cell resolution set the stage for the study of HBV-host cell interaction. These studies revealed that the QLDPAF epitope of the HBV surface antigen large protein (LHBsAg) plays a major role in this process. Quantitative measurements suggested the presence of a second player in this process and both act synergistically to improve cell attachment. As the step of virus-cell attachment is potentially susceptible to specific inhibitors, understanding the molecular basis of virus-cell attachment can be expected to have therapeutic impacts. PMID- 12740830 TI - Hepatitis E virus. AB - Hepatitis E virus (HEV) is a positive-stranded RNA virus with a 7.2 kb genome that is capped and polyadenylated. The virus is currently unclassified: the organisation of the genome resembles that of the Caliciviridae but sequence analyses suggest it is more closely related to the Togaviridae. Hepatitis E virus is an enterically transmitted virus that causes both epidemics and sporadic cases of acute hepatitis in many countries of Asia and Africa but only rarely causes disease in more industrialised countries. Initially the virus was believed to have a limited geographical distribution. However, serological studies suggest that HEV may be endemic also in the United States and Europe even though it infrequently causes overt disease in these countries. Many different animal species worldwide recently have been shown to have antibodies to HEV suggesting that hepatitis E may be zoonotic. Although two related strains have been experimentally transmitted between species, direct transmission from an animal to a human has not been documented. There are four currently recognised genotypes and two of the four contain viruses isolated from swine as well as from humans. Regardless of country of origin or genotype of the virus, most, if not all, strains belong to a single serotype. A promising recombinant vaccine candidate comprised of a truncated capsid protein is currently under evaluation in Nepal. PMID- 12740831 TI - The impact of adenovirus infection on the immunocompromised host. AB - Adenovirus (Ad) infections in immunocompromised hosts have increased in frequency as the number of patients with transplants of bone marrow, liver, kidney, heart and other organs increase in number and survive longer. The numbers of such patients have also increased because of the emergence of the HIV epidemic. Ad infections with the 51 different serotypes recognised to date have few pathognomonic signs and symptoms, and thus require a variety of laboratory-based procedures to confirm infection. These viruses have the ability to target various organs with relative serotype specificity and can cause diverse manifestations including serious life-threatening diseases characteristic of the organs involved. Ads have cytolytic and immunoregulatory properties. The clinical dilemma remains the prompt recognition of Ad-related disease, the differentiation of Ad infection from Ad disease and the differentiation from other causative agents. Since the armamentarium of effective antiviral agents available to treat Ads is unproven by controlled trials and the virus is often not acquired de novo, it is difficult to prevent reactivation in immunodeficient hosts or new acquisition from donor organs. Timely discontinuation of immunosuppressive agents is necessary to prevent morbid outcomes. The clinical diseases, diagnostic tests, antiviral agents and biological aspects of the Ads as pathogens in immunocompromised patients are discussed in the context of this review. Some of the newer diagnostic tests are based on the well-studied molecular biology of Ads, which also have been attenuated by selective viral DNA deletions for use as vectors in numerous gene therapy trials in humans. PMID- 12740833 TI - Neurological manifestations of human parvovirus B19 infection. AB - Since its discovery, human parvovirus B19 has been linked with a broad spectrum of clinical syndromes. An aetiological role for the virus has been confirmed in erythema infectiosum, transient aplastic crisis, persistent infection manifesting as pure red cell aplasia in immunocompromised persons, non-immune hydrops fetalis and arthritis. Less commonly recognised, but receiving increasing attention recently, are the neurological manifestations, a variety of which have been described in patients with either clinically diagnosed or laboratory confirmed B19 infection. The purpose of this review is to summarise present knowledge of B19, its known and potential pathogenic mechanisms and its association with human diseases, particularly those with neurological manifestations. The outcome of the review supports an aetiological role of the virus in neurological disease. However, the pathogenesis remains unknown and elucidating this is a priority. PMID- 12740836 TI - Rational chemical strategies for carbon nanotube functionalization. AB - Whereas the chemistry of fullerenes is well-established, the chemistry of single walled carbon nanotubes (SWNTs) is a relatively unexplored field of research. Investigations into the bonding of moieties onto SWNTs are important because they provide fundamental structural insight into how nanoscale interactions occur. Hence, understanding SWNT chemistry becomes critical to rational, predictive manipulation of their properties. Among the strategies discussed include molecular metal complexation with SWNTs to control site-selective chemistry in these systems. In particular, work has been performed with Vaska's and Wilkinson's complexes to create functionalized adducts. Functionalization should offer a relatively simple means of tube solubilization and bundle exfoliation, and also allows for tubes to be utilized as recoverable catalyst supports. Solubilization of oxidized SWNTs has also been achieved through derivatization by using a functionalized organic crown ether. The resultant adduct yielded concentrations of dissolved nanotubes on the order of 1 g L(-1) in water and at elevated concentrations in a range of organic solvents, traditionally poor for SWNT manipulation. To further demonstrate chemical processability of SWNTs, we have subjected them to ozonolysis, followed by treatment with various independent reagents, to rationally generate a higher proportion of oxygenated functional groups on the nanotube surface. This protocol has been found to purify nanotubes. More importantly, the reaction sequence has been found to ozonize the sidewalls of these nanotubes. Finally, SWNTs have also been chemically modified with quantum dots and oxide nanocrystals. A composite heterostructure consisting of nanotubes joined to nanocrystals offers a unique opportunity to obtain desired physical, electronic, and chemical properties by adjusting synthetic conditions to tailor the size and structure of the individual sub-components, with implications for self-assembly. PMID- 12740832 TI - Split genes and their expression in Kaposi's sarcoma-associated herpesvirus. AB - A split or interrupted gene is defined as a gene consisting of introns and exons. Removal (splicing) of the intron(s) from a primary transcript (pre-mRNA) is essential for creating a mRNA. Initial assignment of a potential protein coding region in the KSHV genome was based on the initiation codon context and predicted protein size larger than 100 amino acids, but the gene discontinuity was disregarded. Experimental investigation of the assigned ORFs has demonstrated that there are up to 25 split genes, more than one fourth of the total KSHV genes described in the KSHV genome. This includes the genes involved in all phases (latent, immediate early, early and late) of KSHV infection. The complexity of a split gene expression depends upon the availability of a proximal promoter and polyadenylation (pA) signal. Sharing a single promoter or a single pA signal by two or three genes is not uncommon in the expression of KSHV split genes and the resulting transcripts are usually polycistronic. Among those of KSHV split genes, 15 genes express a bicistronic or tricistronic RNA and 10 genes express a monocistronic RNA. Alternative RNA splicing could happen in a particular pre-mRNA due to intron or exon inclusion or skipping or the presence of an alternative 5' splice site or 3' splice site. This may, respectively, result in at least 8 species of K8 and 14 species of K15 transcripts. This appears to be related to cell differentiation and stages of the virus infection, presumably involving viral cis elements and trans splicing factors. PMID- 12740838 TI - Pyrene-derived novel one- and two-component organogelators. AB - A new class of alkyl-chain-appended pyrene derivatives 4-14 were synthesized and evaluated for their gelation abilities. Depending on the nature of the linking group, these compounds gelated a number of organic solvents, either in the presence or in the absence of the acceptor molecule 2,4,7-trinitrofluorenone (TNF). Compounds with ester, ether, or alkyl linkages gelated a number of hydroxylic and hydrocarbon solvents by means of a charge-transfer interaction with TNF, while compounds with amide, urethane and urea linkers formed gels on their own in a variety of solvents by means of pi-pi stacking and hydrogen bonding interactions. The Xray crystal structure of urethane (S)-12 showed hydrogen-bonding and stacking features, as suggested by the model. The gels obtained were investigated by spectroscopic and electron microscopic techniques which provided structural insights. PMID- 12740837 TI - Gold glyconanoparticles: synthetic polyvalent ligands mimicking glycocalyx-like surfaces as tools for glycobiological studies. AB - A simple and versatile methodology is described for tailoring sugar functionalised gold nanoclusters (glyconanoparticles) that have 3D polyvalent carbohydrate display and globular shapes. This methodology allows the preparation of glyconanoparticles with biologically significant oligosaccharides as well as with differing carbohydrate density. Fluorescent glyconanoparticles have been also prepared for labelling cells in biological tests. The materials are water soluble, stable under physiological conditions and present an exceptional small core size. All of them have been characterised by (1)H NMR, UV and IR spectroscopy, TEM and elemental analysis. Their highly polyvalent network can mimic glycosphingolipid clustering and interactions at the plasma membrane, providing an controlled system for glycobiological studies. Furthermore, they are useful building blocks for the design of nanomaterials. PMID- 12740839 TI - Mutations in distant residues moderately increase the enantioselectivity of Pseudomonas fluorescens esterase towards methyl 3bromo-2-methylpropanoate and ethyl 3phenylbutyrate. AB - Directed evolution combined with saturation mutagenesis identified six different point mutations that each moderately increases the enantioselectivity of an esterase from Pseudomonas fluorescens (PFE) towards either of two chiral synthons. Directed evolution identified a Thr230Ile mutation that increased the enantioselectivity from 12 to 19 towards methyl (S)-3-bromo-2-methylpropanoate. Saturation mutagenesis at Thr230 identified another mutant, Thr230Pro, with higher-than-wild-type enantioselectivity (E=17). Previous directed evolution identified mutants Asp158Asn and Leu181Gln that increased the enantioselectivity from 3.5 to 5.8 and 6.6, respectively, towards ethyl (R)-3-phenylbutyrate. In this work, saturation mutagenesis identified other mutations that further increase the enantioselectivity to 12 (Asp158Leu) and 10 (Leu181Ser). A homology model of PFE indicates that all mutations lie outside the active site, 12-14 A from the substrate and suggests how the distant mutations might indirectly change the substrate-binding site. Since proteins contain many more residues far from the active site than close to the active site, random mutagenesis is strongly biased in favor of distant mutations. Directed evolution rarely screens all mutations, so it usually finds the distant mutations because they are more common, but probably not the most effective. PMID- 12740840 TI - Hydrogen-hydrogen bonding: a stabilizing interaction in molecules and crystals. AB - Bond paths linking two bonded hydrogen atoms that bear identical or similar charges are found between the ortho-hydrogen atoms in planar biphenyl, between the hydrogen atoms bonded to the C1-C4 carbon atoms in phenanthrene and other angular polybenzenoids, and between the methyl hydrogen atoms in the cyclobutadiene, tetrahedrane and indacene molecules corseted with tertiary-tetra butyl groups. It is shown that each such H-H interaction, rather than denoting the presence of "nonbonded steric repulsions", makes a stabilizing contribution of up to 10 kcal mol(-1) to the energy of the molecule in which it occurs. The quantum theory of atoms in molecules-the physics of an open system-demonstrates that while the approach of two bonded hydrogen atoms to a separation less than the sum of their van der Waals radii does result in an increase in the repulsive contributions to their energies, these changes are dominated by an increase in the magnitude of the attractive interaction of the protons with the electron density distribution, and the net result is a stabilizing change in the energy. The surface virial that determines the contribution to the total energy decrease resulting from the formation of the H-H interatomic surface is shown to account for the resulting stability. It is pointed out that H-H interactions must be ubiquitous, their stabilization energies contributing to the sublimation energies of hydrocarbon molecular crystals, as well as solid hydrogen. H-H bonding is shown to be distinct from "dihydrogen bonding", a form of hydrogen bonding with a hydridic hydrogen in the role of the base atom. PMID- 12740841 TI - Theoretical evidence of persistent chirality in D3 homoleptic hexacoordinate complexes with monodentate ligands. AB - A theoretical study of the enantiomer interconversion pathway relevant to racemization reactions of hexacoordinate transition-metal complexes is presented based on density functional calculations. The potential-energy surface for the trigonal twist pathway of the [Zr(SH)(6)](2-) model compound has been explored. The optimum structure reproduces, to a very good approximation, the experimental geometry of the analogous compound in which the thiolato groups have C(6)H(4)-4 OMe substituents instead of H atoms. A barrier of about 19 kcal mol(-1) is estimated for the racemization of [Zr(SH)(6)](2-) and exploratory calculations for [Zr(SC(6)H(4)-4-OMe)(6)](2-) indicate that a larger barrier should be expected. For the chiral homoleptic organometallic complexes [ZrMe(6)](2-) and [RhMe(6)](3-) no significant racemization barrier is expected. PMID- 12740843 TI - Solution- and bound-state conformational study of N,N',N"-triacetyl chitotriose and other analogous potential inhibitors of hevamine: application of trNOESY and STD NMR spectroscopy. AB - The solution-state conformations of N,N',N"-triacetyl chitotriose (1) and other potential chitinase inhibitors 2-4 were studied using a combination of NMR spectroscopy (NOESY) and molecular mechanics calculations. Determination solely of the global energy minimum conformation was found to be insufficient for an agreement with the NMR results. An appropriate consistency between the NMR experimental data and theoretical calculations was only reached by assessing the structures as population-weighted average conformers based on Boltzmann distributions derived from the calculated relative energies. Analogies, but also particular differences, between the synthetic compounds 2-4 and the naturally occurring N,N',N"-triacetyl chitotriose were found. Furthermore, the conformation of compounds 1 and 2 when bound to hevamine was also studied using transferred NOESY experiments and the binding process was found to impart a level of conformational restriction on the ligands. The preferred conformation as determined for 1 in the bound state to hevamine belonged to one of the conformational families found for the compound when free in solution, although full characterisation of the bound-state conformations was impeded due to severe signal overlap. Saturation transfer difference NMR experiments were also employed to analyse the binding epitopes of the bound compounds. We thus determined that it is mainly the acetyl amido groups of the trisaccharide and the heterocyclic moiety which are in close contact with hevamine. PMID- 12740844 TI - Mechanisms of electrochemically-induced retro-cyclopropanation reactions of fullerene derivatives using digital simulations. AB - Three C(60) derivatives, 1, 2 and 3, have been studied by cyclic voltammetry (CV) under high vacuum in anhydrous tetrahydrofuran (THF). The CV behavior was essentially similar to that already observed for other cyclopropanated fullerene derivatives. After the second reduction processes all compounds undergo a chemical reaction that generates another electroactive species. This "new" chemical species is likely to be the compound with the cyclopropane ring open. Differences in CV behavior were observed for the different addends. Electrochemical data obtained at different scan rates for a given potential window, were fit with the BAS digital simulation program, DigiSim. The purpose of this study was to probe the proposed mechanisms and to obtain reliable estimations of the kinetic constants for the homogeneous chemical reactions taking place during the CV experiments. Calculations at the PM3 level lend additional support to the conclusions derived from digital simulations. The proposed mechanism is similar for all the compounds and involves two main chemical reactions in a reversible square scheme. PMID- 12740842 TI - Novel and anti-inflammatory constituents of Garcinia subelliptica. AB - Four novel phloroglucinol derivatives, garcinielliptones A (1), B (2), C (3), D (4), a novel triterpenoid, garcinielliptone E (5), and three known compounds were isolated from the seeds of Garcinia subelliptica. The structures, including relative configurations, were elucidated by means of spectroscopic data. Known compounds garsubellin A (6) and garcinielliptin oxide (7) showed potent inhibitory effects on the release of beta-glucuronidase, and beta-glucuronidase and histamine, respectively, from peritoneal mast cells stimulated with compound 48/80 in a concentration-dependent manner with IC(50) values of 15.6+/-2.5, and 18.2+/-3.6 and 20.0+/-2.7 microM, respectively. Compound 7 showed potent inhibitory effects on the release of beta-glucuronidase and lysozyme from neutrophils stimulated with formyl-Met-Leu-Phe(fMLP)/cytochalasin B (CB) in a concentration-dependent manner with IC(50) values of 15.7+/-3.0 and 23.9+/-3.2 microM, respectively. Compound 7 also showed potent inhibitory effect on superoxide formation from neutrophils stimulated with fMLP/CB also in a concentration-dependent manner with an IC(50) value of 17.9+/-1.5 microM. PMID- 12740845 TI - Coordination chemistry of the soluble metal oxide analogue [Mo5O13(OCH3)4(NO)]3- with manganese carbonyl species. AB - The reactions of neutral or cationic manganese carbonyl species towards the oxo nitrosyl complex [Na(MeOH)[Mo(5)O(13)(OCH(3))(4)(NO)]](2-) have been investigated in various conditions. This system provides an unique opportunity for probing the basic reactions involved in the preparation of solid oxide-supported heterogeneous catalysts, that is, mobility of transition-metal species at the surface and dissolution-precipitation of the support. Under nitrogen and in the dark, the reaction of in situ generated fac-[Mn(CO)(3)](+) species with (nBu(4)N)(2)[Na(MeOH)-[Mo(5)O(13)(OMe)(4)(NO)]] in MeOH yields (nBu(4)N)(2)[Mn(CO)(3)(H(2)O)[Mo(5)O(13)(OMe)(4)(NO)]] at room temperature, while (nBu(4)N)(3)[Na[Mo(5)O(13)(OMe)(4)(NO)](2)[Mn(CO)(3)](2)] is obtained under reflux. The former transforms into the latter under reflux in methanol in the presence of sodium bromide; this involves the migration of the fac-[Mn(CO)(3)](+) moiety from a basal kappa(2)O coordination site to a lateral kappa(3)O site. Oxidation and decarbonylation of manganese carbonyl species as well as degradation of the oxonitrosyl starting material and reaggregation of oxo(methoxo)molybdenum fragments occur in non-deareated MeOH, and both (nBu(4)N)(4)[Mn(H(2)O)(2)[Mo(5)O(16)(OMe)(2)](2)[Mn(CO)(3)](2)] and (nBu(4)N)(4)[Mn(H(2)O)(2)[Mo(5)O(13)(OMe)(4)(NO)](2)] as well as (nBu(4)N)(2)[MnBr[Mo(5)O(13)(OMe)(4)(NO)]] have been obtained in this way. The rhenium analogue (nBu(4)N)(2)[Re(CO)(3)(H(2)O)[Mo(5)O(13)(OMe)(4)(NO)]] has also been synthesized. The crystal structures of (nBu(4)N)(2)[Re(CO)(3)(H(2)O)[Mo(5)O(13)(OMe)(4)(NO)]], (nBu(4)N)(3)[Na[Mo(5)O(13)(OMe)(4)(NO)](2)[Mn(CO)(3)](2)], (nBu(4)N)(4)[Mn(H(2)O)(2)[Mo(5)O(16)(OMe)(2)](2)[Mn(CO)(3)](2)], (nBu(4)N)(4)[Mn(H(2)O)(2)[Mo(5)O(13)(OMe)(4)(NO)](2)] and (nBu(4)N)(2)[MnBr[Mo(5)O(13)(OMe)(4)(NO)]] have been determined. PMID- 12740846 TI - A distinctive example of the cooperative interplay of structure and environment in tuning of intramolecular charge transfer in second-order nonlinear optical chromophores. AB - The strongly enhanced cooperative influence of medium polarity and organic structural design on the first hyperpolarizability beta of a novel family of highly polarizable azinium-(CH=CH-thienyl)-dicyanomethanido chromophores 1-3 is described. The dyes can be efficiently synthesized by regioselective protonation/alkylation of the corresponding bidentate anion precursors. Consecutive annelation of the pyridyl ring of 1 (pyridine-->quinoline-->acridine) and medium polarity effects are responsible for an extraordinarily variable range of intramolecular charge transfer (ICT), leading to a large set of pi-electron distribution patterns. Accordingly, systems with remarkably different zwitterionic/quinoid character in the ground and excited states present beta values in a broad range, eventually switching from negative to positive. Our investigation is based on a combination of experimental (UV/Vis spectroscopy, multinuclear NMR spectroscopy, and electrooptical absorption measurements) and computational (ab initio) approaches. It is shown that: 1) beta and mubeta are dramatically influenced, even by orders of magnitude, by a complex, non-monotonic interplay of structure and medium action, which in turn affects molecular ICT and bond length alternation (BLA), 2) the computations, validated by different experimental data, are to be recommended as an extremely useful tool in the search for a greatly improved set of molecular nonlinear optical (NLO) responses (in the case of 1-3 they show that such conditions may be attained only in a narrow and limited range of dielectric constants in which the annelation effect operates most efficiently), and 3) the search for the most favorable molecular NLO response of a highly polarizable chromophore both in solution and in solid matrices should simultaneously take into account not only the molecular design supplemented by annelation effects but also the polarity of the medium. PMID- 12740847 TI - Formation of novel sulfur-containing C60F16 cycloadducts between tetrathiafulvalene and C60F18; a unique six-electron cycloaddition of a fullerene involving F2 loss. AB - Co-evaporation of solutions of C(60)F(18) and tetrathiafulvalene in toluene produces an unsymmetrical C(60)F(16):tetrathiafulvalene adduct through a unique six-electron cycloaddition involving displacement of two fluorine atoms by a terminal Cdbond;C double bond of the fulvalene. The adduct rearranges into two further adducts, one of which is characterised as a new type of fullerene derivative, a thiiranofullerene, formed by elimination of a thioketene moiety from the tetrathiafulvalene adduct. The initial addition also produces a bisadduct in which the addends comprise one tetrathiafulvalene molecule and one in which carbon disulfide has been eliminated. The latter adduct involves cycloaddition of an unsaturated aromatic dithiolactone moiety. PMID- 12740848 TI - Palladium(II) complexes of the reducing sugars D-arabinose, D-ribose, rac mannose, and D-galactose. AB - The cellulose solvent Pd-en, an aqueous solution of [(en)Pd(II)(OH)(2)] (en=ethylenediamine), reacts with the monosaccharides D-arabinose (D-Ara), D ribose (D-Rib), rac-mannose (rac-Man), and D-galactose (D-Gal) under formation of dimetalated aldose complexes, if the molar ratio of Pd and sugar is 2:1 or larger. In the Pd(2) complexes, the aldoses are tetra-deprotonated and act as bisdiolato ligands. Two crystalline pentose complexes were isolated: [(en)(2)Pd(2)(beta-D-Arap1,2,3,4 H(-4))].5 H(2)O (1) and [(en)(2)Pd(2)(beta-D Ribp1,2,3,4 H(-4))].6.5 H(2)O (2), along with two hexose complexes. With rac-Man, the major solution species is crystallized as the 9.4-hydrate [(en)(2)Pd(2)(beta rac-Manp1,2,3,4 H(-4))].9.4 H(2)O (3). From the respective D-Gal solutions, [(en)(2)Pd(2)(beta-D-Galf1,3,5,6 H(-4))].5 H(2)O.C(2)H(5)OH (4), with the sugar tetraanion in its furanose form, is crystallized though it is not the major species, rather the second most abundant in purely aqueous solutions. The Galf species is enriched in the mother liquors to the extent of 25 % of total sugar content. Substitution of the en ligand by two molecules of ammonia, methylamine, or isopropylamine, respectively, results in the formation of different solution species. With the bulkiest ligand, isopropylamine, monometalation of the aldoses in the 1,2-position is exclusively observed. PMID- 12740849 TI - Syntheses and structures of magnesium pyridine thiolates--model compounds for magnesium binding in photosystem I. AB - Novel magnesium pyridine-2-thiolates were prepared by using alkane elimination chemistry. The resulting complexes display a metal coordination environment composed of sulfur/nitrogen bonding from the intramolecularly stabilized mercaptopyridine ligand, in addition to coordination by the oxygen centers from two THF donors. The compounds are well-suited model compounds for the magnesium centers in Photosystem I, in which magnesium, situated in the central chlorophyll ligand, is bound to sulfur from a nearby methionine residue. All compounds were characterized by (1)H, (13)C NMR, and IR spectroscopy, in addition to Xray crystallography. PMID- 12740850 TI - Constant-distance mode scanning electrochemical microscopy (SECM)--Part I: Adaptation of a non-optical shear-force-based positioning mode for SECM tips. AB - A non-optical shear-force-based detection scheme for accurately controlling the tip-to-sample distance in scanning electrochemical microscopy (SECM) is presented. With this approach, the detection of the shear force is accomplished by mechanically attaching a set of two piezoelectric plates to the scanning probe. One of the plates is used to excite the SECM tip causing it to resonate, and the other acts as a piezoelectric detector of the amplitude of the tip oscillation. Increasing shear forces in close proximity to the sample surface lead to a damping of the vibration amplitude and a phase shift, effects that are registered by connecting the detecting piezoelectric plate to a dual-phase analogue lock-in amplifier. The shear force and hence distance-dependent signal of the lock-in amplifier is used to establish an efficient, computer-controlled closed feedback loop enabling SECM imaging in a constant-distance mode of operation. The details of the SECM setup with an integrated piezoelectric shear force distance control are described, and approach curves are shown. The performance of the constant-distance mode SECM with a non-optical detection of shear forces is illustrated by imaging simultaneously the topography and conductivity of an array of Pt-band microelectrodes. PMID- 12740852 TI - Relations between synthesis and microstructural properties of copper/zinc hydroxycarbonates. AB - Cu/Zn Hydroxycarbonates obtained by co-precipitation of Cu(2+) and Zn(2+) with Na(2)CO(3) have been investigated regarding phase formation and thermal decomposition in two series with varying Cu/Zn ratios prepared according to the decreasing pH and constant pH method. Hydrozincite, aurichalcite and (zincian) malachite were found to form at differing Cu/Zn ratios for both series. For the constant pH preparation the Cu/Zn ratio in zincian-malachite was close to the nominal values whereas excess values were found for the decreasing pH samples. The degree of crystallinity as well as the thermal decomposition temperatures were lower for the constant pH series. All samples containing aurichalcite revealed an unexpected decomposition step at high temperatures evolving exclusively CO(2). The differences in composition and microstucture were traced back to the different pathways of solid formation for the two preparation methods. Substantial changes were observed during the post-precipitation processes of ageing and washing. The effects were studied in detail on samples with a cation ratio of Cu/Zn 70:30 mol %. Ageing of the precipitates in their own solutions is accompanied by a spontaneous crystallization of the initially amorphous solids. The decreasing pH sample develops from a hydroxy-rich material comprising basic copper nitrate (gerhardtite) as an intermediate. Only small changes in the chemistry of the samples were detected for the constant pH precipitation. The findings are summarised into a scheme of solid formation processes that explains the phenomenon of a "chemical memory" of the precipitates when they are converted into Cu/ZnO model catalysts. PMID- 12740851 TI - 1H and 13C NMR characterization of hemiamidal isoniazid-NAD(H) adducts as possible inhibitors of InhA reductase of Mycobacterium tuberculosis. AB - Isoniazid (INH) is easily oxidized with manganese(III) pyrophosphate, a chemical model of the KatG protein involved in activation of INH inside the bacteria Mycobacterium tuberculosis. Performed in the presence of NAD(+), this oxidation generates a family of isomeric INH-NAD(H) adducts, which have been shown to be effective inhibitors of InhA, an enzyme essential in mycolic acid biosynthesis. In this work, we fully characterized by (1)H and (13)C NMR spectroscopy four main species of INH-NAD(H) adducts that coexist in solution. Two of them are open diastereoisomers consisting of the covalent attachment of the isonicotinoyl radical at position four of the nicotinamide coenzyme. The other two result from a cyclization involving the amide group from the nicotinamide and the carbonyl group from the isonicotinoyl radical to give diastereoisomeric hemiamidals. Although an INH-NAD(H) adduct with a 4S configuration has been characterized within the active site of InhA from Xray crystallography and this bound adduct interpreted as an open form (Rozwarski et al., Science 1998, 279, 98-102), it is legitimate to raise the question about the effective active form(s), open or cyclic, of INH-NAD(H) adduct(s). Is there a single active form or are several forms able to inhibit the InhA activity with different levels of inhibitory potency? PMID- 12740853 TI - N(epsilon) functionalization of metal and organic protected L-histidine for a highly efficient, direct labeling of biomolecules with [Tc(OH2)3(CO)3]+. AB - Two different pathways for the introduction of an acetyl group at N(epsilon ) in a N(alpha), N(delta), and -COO protected histidine to afford N(epsilon) (CH(2)COOH)-histidine derivative 7 b are presented. The purpose of this study is the coupling of 7 b to amino groups in bioactive molecules such as peptides. After full deprotection of such a bioconjugate, histidine provides three coordination sites which efficiently coordinate to [(99m)Tc(OH(2))(3)(CO)(3)](+) or [Re(OH(2))(3)(CO)(3)](+) in a facial geometry. This allows the development of novel radiopharmaceuticals. Selective derivatization at the N(epsilon) position has conveniently been achieved by concomitant protection of N(alpha) and N(delta) with a carbonyl group forming a six-membered urea. Cyclic urea ring opening with Fm-OH, coupling of phenylalanine as a model to 7 b through its primary amine and removing of all protecting groups in one step gave a histidine derivative of phenylalanine which could be labeled at 10(-5) M with (99m)Tc in very high yield and even in about 50 % yield at 10(-6) M. The Xray structure of a complex with [Re(CO)(3)](+) in which anilin is coupled to 7 b confirms the facial arrangement of histidine. A second pathway applies directly the [Re(CO)(3)](+) moiety as a protecting group. This is one of the rare examples in which a metal fragment is used as a protecting group for organic functionalities. The coordination to histidine protects the N(alpha), N(delta) and COO group in one single step, subsequent alkylation with BrCH(2)COOH(R) at N(epsilon), coupling to phenylalanine and oxidative deprotection of [Re(CO)(3)](+) to [ReO(4)](-) gave the corresponding bioconjugate in which histidine is coupled to phenylalanine through an acetylamide at N(epsilon). Both methods offer convenient pathways to introduce histidine in a biomolecule under retention of its three coordination sites. The procedures are adaptable to any biomolecule with pendant amines and allow the development of novel radiopharmaceuticals or inversed peptides. PMID- 12740854 TI - Theoretical study on the mechanism of iron carbonyls mediated isomerization of allylic alcohols to saturated carbonyls. AB - The conversion of allylic alcohols to enols mediated by Fe(CO)(3) has been studied through density functional theoretical calculations. From the results obtained a complete catalytic cycle has been proposed in which the first intermediate is the [(allyl alcohol)Fe(CO)(3)] complex. This intermediate evolves to the [(enol)Fe(CO)(3)] complex through two consecutive 1,3-hydrogen shifts involving a pi-allyl hydride intermediate. The highest Gibbs energy transition state corresponds to the partial decoordination ot the enol ligand prior to the coordination of a new allyl alcohol molecule that regenerates the first intermediate. Alternative processes for the [(enol)Fe(CO)(3)] complex such as [Fe(CO)(3)]-mediated enol-aldehyde transformation and enol isomerization have also been considered. The results obtained show that the former process is unfavourable, whereas the enol isomerization may compete with the enol decoordination step of the catalytic cycle. PMID- 12740855 TI - Cycloadditions of isobenzofuran to a constrained template bearing neighboring dienophiles. AB - A high yielding synthesis of the pentacyclic diene-dione 1 has enabled investigation of its reactivity as a double dienophile in Diels-Alder [4+2] cycloadditions with isobenzofuran, leading to novel and highly symmetrical three sided cavitands 3 and 4. PMID- 12740856 TI - Isomeric alkyl cation/arene complexes in the gas phase. AB - The kinetics and the stereochemistry of the protonation-induced unimolecular isomerization of (S)-(+)-1-D(1)-3-(p-tolyl)butane have been investigated in the gas phase in the 100-160 degrees C range. The process leads to the almost exclusive formation of the relevant meta isomer with complete racemization and partial 1,2-H shift in the migrating sec-butyl group. These results, together with the relevant activation parameters, point to the occurrence of low-energy, tightly bound isomeric sec-butyl cation/toluene complexes of defined structure and stability along the isomerization coordinate. The existence and the eta(1) type structure of these low-energy intermediate species are confirmed by ab initio calculations on closely related systems at the MP2(full)/6-311++G**//HF/6 31+G** level of theory. Their role in the relevant energy surface clearly emerges from the comparison of the present results with those concerning sec-butylation of toluene carried out under comparable experimental conditions. PMID- 12740857 TI - Diastereoselective solution and multipin-based combinatorial array synthesis of a novel class of potent phosphinic metalloprotease inhibitors. AB - The solution-phase synthesis and resolution of new phosphinopeptidic building blocks containing a triple bond was realized in high yields and optical purities (units 3 a-d). The absolute configuration of the target compounds was unambiguously established by NMR studies. A post-assembly diversification strategy of these blocks was developed through 1,3-dipolar cycloaddition of a variety of in situ prepared nitrile oxides. This strategy led to the rapid and efficient diastereoselective preparation of a novel class of isoxazole-containing phosphinic peptides (peptides 5 a-i). Solid-phase version of this strategy was efficiently achieved on multipin solid technology, by developing a new protocol for the coupling of P-unprotected dipeptidic blocks with solid supported amino acids in a quantitative and diastereoselective manner. Optimization of dipolar cycloadditions onto pin-embodied phosphinic peptides allowed the convenient preparation of this new class of pseudopeptides. The crude phosphinic peptides (9 a-k) were obtained in high yields and purity as determined by RP-HPLC. Inhibition assays of some of these peptides revealed that they behave as very potent inhibitors of MMPs, outmatching previously reported phosphinic peptides, in terms of potency (K(i) in the range of few nM). PMID- 12740858 TI - Bonding energetics in alkaline metal alkoxides and phenoxides. AB - The bonding energetics in a variety of alkaline metal, alkoxides and phenoxides, MOR, was investigated based on the corresponding enthalpies of formation in the crystalline state determined by reaction-solution calorimetry. The results obtained at 298.15 K were as follows: Delta(f)H(m)(o)(MOR, cr)/kJ mol(-1) = 382.7+/-1.4 (LiOC(6)H(5)), 513.6+/-2.5 (NaO-nC(6)H(13)), 326.4+/-1.4 (NaOC(6)H(5)), 375.2+/-3.4 (KOCH(3)), 434.5+/-2.7 (KOC(2)H(5)), 467.1+/-5.2 (KO nC(3)H(7)), 459.3+/-2.1 (KO-nC(4)H(9)), 464.6+/-5.7 (KO-tC(4)H(9)), 464.3+/-2.5 (KO-nC(6)H(13)), 333.3+/-3.1 (KOC(6)H(5)), 380.6+/-2.9 (RbOCH(3)), 434.1+/-2.9 (RbOC(2)H(5)), 345.3+/-2.9 (LiOC(6)H(5)), 379.1+/-3.0 (CsOCH(3)), 432.3+/-3.1 (CsOC(2)H(5)), 466.9+/-5.0 (CsO-nC(3)H(7)), 461.3+/-3.5 (CsO-nC(4)H(9)), 461.9+/ 2.5 (CsO-tC(4)H(9)), 349.2+/-1.4 (CsOC(6)H(5)). These results together with revised Delta(f)H(m)(o)(MOR, cr) values from the literature, were used to derive a consistent set of lattice energies for the MOR compounds and discuss general trends in the structure-energetics relationship based on the Kapustinskii equation. PMID- 12740859 TI - Ternary complexes between cationic GdIII chelates and anionic metabolites in aqueous solution: an NMR relaxometric study. AB - The (1)H and (17)O NMR relaxometric properties of two cationic complexes formed by Gd(III) with a macrocyclic heptadentate triamide ligand, L(1), and its Nmethylated analogue, L(2), have been investigated in aqueous media as a function of pH, temperature and magnetic field strength. The complexes possess two water molecules in their inner coordination sphere for which the rate of exchange has been found to be sensibly faster for the Nmethylated derivative and explained in terms of electronic effects (decrease of the charge density at the metal center) and perturbation of the network of hydrogen-bonded water molecules in the outer hydration sphere. The proton relaxivity shows a marked dependence from pH and decreases of about six units in the pH range 6.5 to 9.0. This has been accounted for by the displacement of the two water molecules by dissolved carbonate which acts as a chelating anion. The formation of ternary complexes with lactate, malonate, citrate, acetate, fluoride and hydrogenphosphate has been monitored by (1)H NMR relaxometric titrations at 20 MHz and pH 6.3 and the value of the affinity constant, K, and of the relaxivity of the adducts could be obtained. Lactate, malonate and citrate interact strongly with the complexes (log K > or =3.7) and coordinate in a bidendate mode by displacing both water molecules. Larger affinity constants have been measured for GdL(2). Acetate, fluoride and hydrogenphosphate form monoaqua ternary complexes which were investigated in detail with regard to their relaxometric properties. The NMR dispersion (NMRD) profiles indicate a large contribution to the relaxivity of the adducts from water molecules belonging to the second hydration shell of the complexes and hydrogen-bonded to the anion. A VT (17)O NMR study has shown a marked increase of the rate of water exchange upon binding which is explained by coordination of the anion in an equatorial site, thus leaving the water molecule in an apical position, more accessible for interactions with the solvent molecules of the second hydration shell which facilitate the exchange process. PMID- 12740860 TI - Alternative heterocycles for DNA recognition: the benzimidazole/imidazole pair. AB - Boc-protected benzimidazole-pyrrole, benzimidazole-imidazole, and benzimidazole methoxypyrrole amino acids were synthesized and incorporated into DNA binding polyamides, comprised of N-methyl pyrrole and N-methyl imidazole amino acids, by means of solid-phase synthesis on an oxime resin. These hairpin polyamides were designed to determine the DNA recognition profile of a side-by-side benzimidazole/imidazole pair for the designated six base pair recognition sequence. Equilibrium association constants of the polyamide-DNA complexes were determined at two of the six base pair positions of the recognition sequence by quantitative DNase I footprinting titrations on DNA fragments each containing matched and single base pair mismatched binding sites. The results indicate that the benzimidazole-heterocycle building blocks can replace pyrrole-pyrrole, pyrrole-imidazole, and pyrrole-hydroxypyrrole constructs while retaining relative site specifities and subnanomolar match site affinities. The benzimidazole containing hairpin polyamides represent a novel class of DNA binding ligands featuring tunable target recognition sequences combined with the favorable properties of the benzimidazole type DNA minor groove binders. PMID- 12740861 TI - Improved one-pot synthesis of styryl tetrahydrofurans and cyclohexanes by radical addition to beta-nitrostyrenes in the presence of benzoyl peroxide. AB - Stereoselective styryl derivatives have been prepared based on radical substitution (addition-elimination) of heterocycles or cyclohydrocarbons units to (E)-beta-nitrostyrenes 1 using a common radical initiator benzoyl peroxide. High reactivity and selectivity with wide substrate scope were attained by using this easy methodology. The reactions using easily obtained and one-pot potential starting materials gave excellent trans-selectivity with medium to high yields in all cases. Synthetic utility of this approach has been demonstrated by the preparation of various trans-styryl derivatives. PMID- 12740863 TI - Blockade of neuronal facilitatory nicotinic receptors containing alpha 3 beta 2 subunits contribute to tetanic fade in the rat isolated diaphragm. AB - Nicotinic receptor (nAChR) subtypes involved in pre- and postjunctional actions underlying tetanic fade were studied in rat phrenic-nerve hemidiaphragms. We investigated the ability of subtype-specific nAChR antagonists to depress nerve evoked contractions and [(3)H]-acetylcholine ([(3)H]-ACh) release. Muscle tension was transiently increased during brief high frequency trains (50 Hz for 5 sec). The rank potency order of nAChR antagonists to reduce tetanic peak tension was alpha-bungarotoxin > d-tubocurarine >> mecamylamine > hexamethonium. Reduction of maximal tetanic tension produced by dihydro-beta-erythroidine (0.03-10 microM), methyllycaconitine (0.003-3 microM), and alpha-conotoxin MII (0.001-0.3 microM) did not exceed 30%. Besides reduction of peak tension d-tubocurarine (0.1-0.7 microM), mecamylamine (0.1-300 microM), and hexamethonium (30-3,000 microM) also caused tetanic fading. With alpha-conotoxin MII (0.001-0.3 microM) and dihydro beta-erythroidine (0.03-10 microM), tetanic fade was evident only after decreasing the safety factor of neuromuscular transmission (with high magnesium ions, 6-7 mM). The antagonist rank potency order to reduce evoked (50 Hz for 5 sec) [(3)H]-ACh release from motor nerve terminals was alpha-conotoxin MII (0.1 microM) > dihydro-beta-erythroidine (1 microM) approximately d-tubocurarine (1 microM) > mecamylamine (100 microM) > hexamethonium (1,000 microM). When applied in a concentration (0.3 microM) above that producing tetanic paralysis, alpha bungarotoxin failed to affect [(3)H]-ACh release. Data obtained suggest that postjunctional neuromuscular relaxants interact with alpha-bungarotoxin-sensitive nicotinic receptors containing alpha1-subunits, whereas blockade of neuronal alpha3beta2-containing receptors produce tetanic fade by breaking nicotinic autofacilitation of acetylcholine release. PMID- 12740864 TI - Neurotoxic methamphetamine regimen severely impairs recognition memory in rats. AB - Methamphetamine (mAMPH), when administered repeatedly to rodents or primates, is neurotoxic to some cortical neurons and to forebrain dopaminergic and serotonergic axon terminals. The aim of the present study was to investigate the effects of a neurotoxic regimen of mAMPH on two hippocampus-dependent memory tasks: object recognition, a nonspatial memory task, and the Morris water maze, a spatial memory task. Male rats were treated with mAMPH (4 x 4.0 mg/kg, s.c.) or saline and trained in the object recognition task 1 week and 3 weeks later. During training, animals explored two identical copies of the same object. In retention test sessions one of the objects was replaced by a novel object. mAMPH treated rats showed no recognition memory during the short-term memory (STM) test, given 90 min after the training session, and showed marked impairments in the long-term memory (LTM) test, given 24 h after training. Even 3 weeks after drug injections, the mAMPH-treated animals were unable to discriminate between the novel and familiar objects during both STM and LTM tests. Despite the severe deficits observed in the recognition memory, no effects of prior mAMPH treatment were seen in the water maze task. Damage to monoamine terminals was confirmed by significant 30-40% losses of [(125)I]RTI-55 binding to striatal dopamine transporter and hippocampal serotonin transporter sites at both 1 and 3 weeks after mAMPH treatments. Thus, administration of mAMPH restricted to a single day can produce a profound, persistent, and selective deficit in a nonspatial hippocampus-dependent memory. PMID- 12740865 TI - Monocarboxylate transporters contribute to the adaptation of neuronal activity to repeated glucose deprivation in the rat lateral septal nucleus. AB - Using optical recording methods in the rat lateral septal nucleus (LSN) slice, we examined the question of whether antecedent hypoglycemia protects neurons from the adverse effects of subsequent hypoglycemic stimuli. The first exposure of LSN neurons to glucose deprivation for 15 min produced a marked depression of neuronal activity. The neuronal activity recovered by reapplication of glucose to the neurons. In neurons previously exposed to glucose deprivation, subsequent applications of glucose-free artificial cerebrospinal fluid (ACSF) produced only weak depression of the neuronal activity. The contribution of monocarboxylate transporters to this adaptation of neuronal activity to repeated glucose deprivation was examined in LSN neurons. alpha-Cyano-4-hydroxycinnamate (4-CIN, 100 microM), an inhibitor of the monocarboxylate transporters, did not significantly affect the depression of the neuronal activity induced by the first glucose deprivation. However, in the presence of 4-CIN (100 microM), a second glucose deprivation produced a typical depression of the neuronal activity, indicating that 4-CIN had nullified the adaptation of neuronal activity to a second glucose deprivation. Cytochalasin B (CCB, 20 microM), an inhibitor of glucose transporters, depressed the neuronal activity in the presence of 11 mM glucose. Pyruvate (11 mM) partially restored the neuronal activity depressed by pretreatment with CCB (20 microM) for 30-40 min. These results suggest that antecedent glucose deprivation stimulates monocarboxylate-transporters to supply energy substrates to LSN neurons, thus protecting the neurons against subsequent glucose deprivation. . PMID- 12740866 TI - Role for calbindin-D28K in in vitro classical conditioning of abducens nerve responses in turtles. AB - Intracellular calcium has a pivotal role in synaptic modifications that may underlie learning and memory. The present study examined whether there were changes in immunoreactivity levels of the AMPA receptor subunits GluR2/3 and calcium binding proteins during classical conditioning recorded in the abducens nerve of in vitro brain stem preparations from turtles. The results showed that abducens motor neurons in unconditioned turtle brain stems were immunopositive for GluR2/3, calbindin-D28K, and calmodulin, but were immunonegative for parvalbumin. After classical conditioning, immunoreactivity for calbindin-D28K in the abducens motor nuclei was significantly reduced, whereas there were no significant changes in GluR2/3, calmodulin, or parvalbumin. This reduction in calbindin-D28K immunoreactivity was not observed following conditioning in the NMDA receptor antagonist AP-5, which blocked conditioned responses, suggesting that these changes are NMDA receptor-dependent. Moreover, the degree of the decrease in calbindin-D28K immunoreactivity was negatively correlated with the level of conditioning. Consistent with the immunocytochemical findings, Western blot analysis showed that calbindin-D28K protein levels were reduced after classical conditioning. The results support the hypothesis that in vitro classical conditioning of abducens nerve responses utilizes intracellular calcium dependent signaling pathways that require NMDA receptor function and suggest a specific role for the calcium binding protein calbindin-D28K. PMID- 12740867 TI - Changes in NPY-mediated modulation of hippocampal [3H]D-aspartate outflow in the kindling model of epilepsy. AB - The anticonvulsant effect of NPY may depend on Y(2) and/or Y(5) receptor-mediated inhibition of glutamate release in critical areas, such as the hippocampus. However, Y(2) and Y(5) receptor levels have been reported to increase and decrease, respectively, in the epileptic hippocampus, implicating that the profile of NPY effects may change accordingly. The aim of this study was to evaluate the differential effects of NPY on glutamate release in the normal and in the epileptic hippocampus. Thus, we pharmacologically characterized the effects of NPY on the release of [(3)H]D-aspartate, a valid marker of endogenous glutamate, from synaptosomes prepared from the whole hippocampus and from the three hippocampal subregions (dentate gyrus and CA1 and CA3 subfields) of control and kindled rats, killed 1 week after the last stimulus-evoked seizure. In the whole hippocampus, NPY does not significantly affect stimulus-evoked [(3)H]D aspartate overflow. In synaptosomes prepared from control rats, NPY significantly inhibited 15 mM K(+)-evoked [(3)H]D-aspartate overflow only in the CA1 subfield (approx. -30%). Both Y(2) and Y(5) receptor antagonists (respectively, 1 microM BIIE0246 and 1 microM CGP71683A) prevented this effect, suggesting the involvement of both receptor types. In contrast, in synaptosomes prepared from kindled rats NPY significantly inhibited 15 mM K(+)-evoked [(3)H]D-aspartate overflow in the CA1 subfield and in the dentate gyrus (approx. -30%). Only the Y(2) (not the Y(5)) antagonist prevented these effects. These data indicate a critical role for the Y(2) receptor in the inhibitory control of glutamate release in the kindled hippocampus and, thus, suggest that the anticonvulsant effect of NPY in the epileptic brain is most likely Y(2), but not Y(5), receptor mediated. PMID- 12740868 TI - Differential effects of amphetamine transport vs. dopamine reverse transport on particulate PKC activity in striatal synaptoneurosomes. AB - Amphetamine has been shown to increase striatal particulate protein kinase C (PKC) activity [Giambalvo (1992b) Neuropharmacology 31:1211-1222]. The present study examined possible mechanisms involved. Specifically, the effects of calcium, endogenous DA, and DA receptors on the amphetamine-induced increase in PKC activity in striatal synaptoneurosomes were examined. Naive rats and rats pretreated with N-ethoxy-carbonyl-2-ethoxy-1,2-dihydroquinolone (EEDQ, i.p.), a nonselective irreversible receptor antagonist, or with alpha-methyl-p-tyrosine (AMPT, i.p.), a DA synthesis inhibitor, were sacrificed and striatal synaptoneurosomes were prepared. The tissue was incubated with amphetamine, with and without calcium, and PKC activity was then determined by the endogenous phosphorylation of endogenous substrate proteins, as described previously [Giambalvo (1988a) Biochem Pharmacol 37:4001-4007]. It was found that calcium enhanced the effect of amphetamine on PKC activity, even in rats pretreated with EEDQ. Intracellular calcium was required since pretreatment with 1,2-bis (2 aminophenoxy) ethane-N, N, N, N-tetracetic acid acetoxymethyl ester (BAPTA-AM) in vitro attenuated the amphetamine-induced increase in PKC activity, resulting in an inhibition of PKC activity instead. Likewise, endogenous DA was essential since pretreatment with AMPT resulted in a similar amphetamine-induced inhibition of PKC activity. Pretreatment with AMPT did not alter the inhibitory effect of the D2 DA agonist, LY 171555, on PKC activity. It did, however, abolish the calcium-dependent stimulatory effect of the D1 agonist SKF 38393 on PKC activity, rendering it inhibitory regardless of calcium. Considering that both BAPTA-AM and AMPT pretreatments, which diminished DA release without affecting uptake via different mechanisms, produced similar inhibitory effects on PKC activity by amphetamine, these results suggest that the inward transport of amphetamine had an inhibitory effect on PKC activity. In contrast, the outward transport of DA seemed to have a stimulatory effect on PKC activity since incubation with low sodium or with ouabain, conditions that promote DA reverse-transport, increased PKC activity. These results showed that PKC activity was altered differently during inward vs. outward transport. The amphetamine-induced increase in PKC activity was attenuated by pretreatment with DA uptake blockers (nomifensine, GBR 12935, and bupropion), even though these drugs by themselves also increased PKC activity. This effect was diminished by calcium and persisted in rats pretreated with EEDQ. Thus, calcium had a differential effect on the PKC activity induced by a transported substrate (amphetamine) vs. nontransported inhibitors (DA uptake blockers). PMID- 12740869 TI - Sensitivities of benzodiazepine receptor binding and muscarinic acetylcholine receptor binding for the detection of neural cell death caused by sodium nitroprusside microinjection in rat brain. AB - Sodium nitroprusside (SNP) was microinjected into rat cerebral cortex and changes in muscarinic acetylcholine receptor (mAChR) binding and benzodiazepine receptor (BZR) binding were followed for 24 h after the infusion using [(3)H]-N-methyl-4 piperidyl benzilate ([(3)H]-NMPB) and [(3)H]-flumazenil, respectively, as a radioligand. The microinjection of SNP dose-dependently caused significant neural cell death 3 h after infusion, with the area of cell death becoming extensive 24 h after infusion. Neither SIN-1 nor NOC-18, other types of NO donors, caused neural cell death. Together with the result that deferoxamine, an iron-chelating agent, protected SNP-induced brain injury indicated important roles of iron related radicals in SNP cytotoxicity in rat brain. In vitro [(3)H]-NMPB binding was significantly reduced in parallel with the time course of neural cell death detected by TTC staining and Nissl staining. In contrast, [(3)H]-flumazenil binding was essentially unaltered during the 24-h period after the SNP infusion. Similar results were observed in in vivo binding experiments. In vivo [(3)H]-NMPB binding was found to be much more sensitive at detecting cell death caused by SNP. On the other hand, [(3)H]-flumazenil binding in vivo was relatively insensitive to SNP-induced cell death. These results indicate that mAChR binding may be superior to BZR binding for detecting cell death in brain tissue, in contrast to what was previously thought. PMID- 12740870 TI - Class III beta-tubulin in human development and cancer. AB - The differential cellular expression of class III beta-tubulin isotype (betaIII) is reviewed in the context of human embryological development and neoplasia. As compared to somatic organs and tissues, betaIII is abundant in the central and peripheral nervous systems (CNS and PNS) where it is prominently expressed during fetal and postnatal development. As exemplified in cerebellar and sympathoadrenal neurogenesis, the distribution of betaIII is neuron-associated, exhibiting distinct temporospatial gradients according to the regional neuroepithelia of origin. However, transient expression of this protein is also present in the subventricular zones of the CNS comprising putative neuronal- and/or glial precursor cells, as well as in Kulchitsky neuroendocrine cells of the fetal respiratory epithelium. This temporally restricted, potentially non-neuronal expression may have implications in the identification of presumptive neurons derived from embryonic stem cells. In adult tissues, the distribution of betaIII is almost exclusively neuron-specific. Altered patterns of expression are noted in cancer. In "embryonal"- and "adult-type" neuronal tumors of the CNS and PNS, betaIII is associated with neuronal differentiation and decreased cell proliferation. In contrast, the presence of betaIII in gliomas and lung cancer is associated with an ascending histological grade of malignancy. Thus, betaIII expression in neuronal tumors is differentiation-dependent, while in non-neuronal tumors it is aberrant and/or represents "dedifferentiation" associated with the acquisition of progenitor-like phenotypic properties. Increased expression in various epithelial cancer cell lines is associated with chemoresistance to taxanes. Because betaIII is present in subpopulations of neoplastic, but not in normal differentiated glial or somatic epithelial cells, the elucidation of mechanisms responsible for the altered expression of this isotype may provide insights into the role of the microtubule cytoskeleton in tumorigenesis and tumor progression. PMID- 12740871 TI - Myosin localization during meiosis I of crane-fly spermatocytes gives indications about its role in division. AB - We showed previously that in crane-fly spermatocytes myosin is required for tubulin flux [Silverman-Gavrila and Forer, 2000a: J Cell Sci 113:597-609], and for normal anaphase chromosome movement and contractile ring contraction [Silverman-Gavrila and Forer, 2001: Cell Motil Cytoskeleton 50:180-197]. Neither the identity nor the distribution of myosin(s) were known. In the present work, we used immunofluorescence and confocal microscopy to study myosin during meiosis I of crane-fly spermatocytes compared to tubulin, actin, and skeletor, a spindle matrix protein, in order to further understand how myosin might function during cell division. Antibodies to myosin II regulatory light chain and myosin II heavy chain gave similar staining patterns, both dependent on stage: myosin is associated with nuclei, asters, centrosomes, chromosomes, spindle microtubules, midbody microtubules, and contractile rings. Myosin and actin colocalization along kinetochore fibers from prometaphase to anaphase are consistent with suggestions that acto-myosin forces in these stages propel kinetochore fibres poleward and trigger tubulin flux in kinetochore fibres, contributing in this way to poleward chromosome movement. Myosin and actin colocalization at the cell equator in cytokinesis, similar to studies in other cells [e.g., Fujiwara and Pollard, 1978: J Cell Biol 77:182-195], supports a role of actin-myosin interactions in contractile ring function. Myosin and skeletor colocalization in prometaphase spindles is consistent with a role of these proteins in spindle formation. After microtubules or actin were disrupted, myosin remained in spindles and contractile rings, suggesting that the presence of myosin in these structures does not require the continued presence of microtubules or actin. BDM (2,3 butanedione, 2 monoxime) treatment that inhibits chromosome movement and cytokinesis also altered myosin distributions in anaphase spindles and contractile rings, consistent with the physiological effects, suggesting also that myosin needs to be active in order to be properly distributed. PMID- 12740872 TI - Deletion of mdmB impairs mitochondrial distribution and morphology in Aspergillus nidulans. AB - Mitochondria form a dynamic network of interconnected tubes in the cells of Saccharomyces cerevisiae or filamentous fungi such as Aspergillus nidulans, Neurospora crassa, or Podospora anserina. The dynamics depends on the separation of mitochondrial fragments, their movement throughout the cell, and their subsequent fusion with the other parts of the organelle. Interestingly, the microtubule network is required for the distribution in N. crassa and S. pombe, while S. cerevisiae and A. nidulans appear to use the actin cytoskeleton. We studied a homologue of S. cerevisiae Mdm10 in A. nidulans, and named it MdmB. The open reading frame is disrupted by two introns, one of which is conserved in mdm10 of P. anserina. The MdmB protein consists of 428 amino acids with a predicted molecular mass of 46.5 kDa. MdmB shares 26% identical amino acids to Mdm10 from S. cerevisiae, 35% to N. crassa, and 32% to the P. anserina homologue. A MdmB-GFP fusion protein co-localized evenly distributed along mitochondria. Extraction of the protein was only possible after treatment with a non-ionic and an ionic detergent (1% Triton X-100; 0.5% SDS) suggesting that MdmB was tightly bound to the mitochondrial membrane fraction. Deletion of the gene in A. nidulans affected mitochondrial morphology and distribution at 20 degrees C but not at 37 degrees C. mdmB deletion cells contained two populations of mitochondria at lower temperature, the normal tubular network plus some giant, non-motile mitochondria. PMID- 12740873 TI - Higher order structure of the PCM adjacent to the centriole. AB - The centrosome is the major microtubule organizing center in most animal cells. This cytoplasmic organelle consists of two components : a mature centriole (or a pair of centrioles) and a mass of pericentriolar material (PCM). The PCM has been described as either a cloud of material that encases the entire centriole or as a cluster of proteins divided into two subsets, one that adheres to the lateral surface of the centriole and another that extends outward from this region as a cloud of material. In contrast to these protein distribution patterns, we demonstrated in a previous study that a subset of proteins present within the PCM is integrated together to form a tube (PCM tube) with an open and closed end that is duplicated in concert with centrosome duplication. The present study was undertaken to determine if this tubular conformation represents proteins that are confined to the surface of the centriole or if it represents a subset of proteins within the cloud of material that extends outward from the centriole. We document that : (1) the PCM tube represents a portion of the PCM directly associated with the centriole; (2) the PCM tube has a specific and reproducible relationship to the polar structure of the centriole; (3) the tube is a site of cytoplasmic microtubule organization, and has a structure that influences the initial pattern of microtubule assembly within the juxta-centriolar region; and (4) the PCM tube has a structural relationship with respect to the centriole, which allows the simultaneous expression of centriole- and PCM-based functions (e.g., ciliogenesis and cytoplasmic microtubule organization). Based on these findings, we propose a new model of the PCM at the centriole. This model highlights the role played by the proximal end of the centriole in the nucleation and organization of centriole associated PCM, and indicates that the centrosome has an overall polarity in the region of the centriole. PMID- 12740875 TI - Commending the initiative of the microsurgery community. PMID- 12740874 TI - Dynamic association of a tumor amplified kinase, Aurora-A, with the centrosome and mitotic spindle. AB - Aurora-A kinase, also known as STK15/BTAK kinase, is a member of a serine/threonine kinase superfamily that includes the prototypic yeast Ipl1 and Drosophila aurora kinases as well as other mammalian and non-mammalian aurora kinases involved in the regulation of centrosomes and chromosome segregation. The Aurora-A gene is amplified and overexpressed in a wide variety of human tumors. Aurora-A is centrosome-associated during interphase, and binds the poles and half spindle during mitosis; its over-expression has been associated with centrosome amplification and multipolar spindles. GFP-Aurora-A was used to mark centrosomes and spindles, and monitor their movements in living cells. Centrosome pairs labeled with GFP-Aurora-A are motile throughout interphase undergoing oscillations and tumbling motions requiring intact microtubules and ATP. Fluorescence recovery after photobleaching (FRAP) was used to examine the relative molecular mobility of GFP-Aurora-A, and GFP-labeled alpha-tubulin, gamma tubulin, and NuMA. GFP-Aurora-A rapidly exchanges in and out of the centrosome and mitotic spindle (t(1/2) approximately 3 sec); in contrast, both tubulins are relatively immobile indicative of a structural role. GFP-NuMA mobility was intermediate in both interphase nuclei and at the mitotic spindle (t(1/2) approximately 23-30 sec). Deletion mapping identifies a central domain of Aurora A as essential for its centrosomal localization that is augmented by both the amino and the carboxyl terminal ends of the protein. Interestingly, amino or carboxy terminal deletion mutants that maintained centrosomal targeting exhibited significantly slower molecular exchange. Collectively, these studies contrast the relative cellular dynamics of Aurora-A with other cytoskeletal proteins that share its micro-domains, and identify essential regions required for targeting and dynamics. PMID- 12740878 TI - Free osteocutaneous lateral arm flap: anatomy and clinical applications. AB - For many surgeons, the potential to reconstruct skin, fascia, tendon, or bone in a single-stage procedure has made the lateral arm flap the technique of choice for reconstruction of complex defects. The aim of this study was to examine more closely how the humeral bone is supplied by the posterior collateral radial artery. To this end, we dissected 30 cadaver arms to determine the vascular relationship of the lateral arm flap to the humerus. The number of directly supplying vessels, and height to the lateral epicondyle of the humerus, were examined. The reconstructive potential of the osteocutaneous flap in different indications is analyzed in a series of five clinical cases. In all dissected extremities, we found one or two branches of the posterior collateral artery directly and constantly supplying the bone between 2-7 cm proximal to the lateral epicondyle. In five cases, combined defects, including bone, were successfully reconstructed with lateral arm flaps, including vascularized bone. PMID- 12740879 TI - Indications for vascularized free rectus abdominis musculocutaneous flap in oromandibular region in terms of efficiency of anterior rectus sheath. AB - The anterior rectus sheath, efficacious in reconstructive surgery, is used in oromandibular reconstruction with the free rectus abdominis musculocutaneous flap. This study describes reconstruction with this sheath in 20 patients: to preserve only the swallowing function in 10 patients (formation of the bulge of the reconstructed oral floor and prevention of its sinking); to preserve both swallowing and articulation in 5 patients (formation of the bulge of the reconstructed tongue and prevention of its sinking, concurrent with a money-pouch like reconstruction of the tongue, laryngeal suspension, and neuroanastomosis); and to prevent exposure of the reconstruction plate, replacing the resected mandibular continuity in 5 patients. The purpose of reconstruction was achieved in all patients. The vascularized free rectus abdominis musculocutaneous flap with a firm anterior rectus sheath may be the first choice for these types of reconstruction. PMID- 12740880 TI - Two-stage reconstruction of four-digit amputation: distal ulnar artery flap and combined second- and third-toe transfer. AB - We report a case of combined two toe transfer after distal ulnar artery fasciocutaneous flap in a 10-year-old boy who has four-digit amputation. Four years after the operation, satisfactory results were obtained both functionally and cosmetically. PMID- 12740881 TI - Clinical application of ipsilateral C7 nerve root transfer for treatment of C5 and C6 avulsion of brachial plexus. AB - We applied a nerve transfer, using the ipsilateral C7 nerve root to treat the C5 and C6 root avulsion of the brachial plexus. Four patients with C5 and C6 preganglionic injury were operated on with this new technique from 1998-2000. Transfer of the spinal accessory nerve to the suprascapular nerve was simultaneously done in 2 these patients. After a follow-up of 1-2.5 years, the muscle strength of elbow flexors recovered to M4 (Lovett) in all cases, shoulder abduction of >90 degrees with external rotation of 30-40 degrees was gained in two cases, and that of 15-45 degrees with no external rotation in the other two cases. No remarkable impairment was found in all C7-innervated muscles except for decrease of muscle power of 1 grade (Lovett) in the short run. This new technique shows promise as an efficacious and safe treatment for C5 and C6 root avulsion of the brachial plexus. However, it should be applied prudently when incomplete injuries of the lower trunk are involved. PMID- 12740882 TI - Carbon dioxide laser-assisted nerve repair: effect of solder and suture material on nerve regeneration in rat sciatic nerve. AB - In order to further improve and explore the role of lasers for nerve reconstruction, this study was designed to investigate regeneration of sharply transected peripheral nerves repaired with a CO(2) milliwatt laser in combination with three different suture materials and a bovine albumin protein solder as an adjunct to the welding process. Unilateral sciatic nerve repair was performed in 44 rats. In the laser group, nerves were gently apposed, and two stay sutures (10 0 nylon, 10-0 polyglycolic acid, or 25 microm stainless steel) were placed epi/perineurially. Thereafter, the repair site was fused at 100 mW with pulses of 1.0 s. In the subgroup of laser-assisted nerve repair (LANR), albumen was used as a soldering agent to further reinforce the repair site. The control group consisted of nerves repaired by conventional microsurgical suture repair (CMSR), using 4-6 10-0 nylon sutures. Evaluation was performed at 1 and 6 weeks after surgery, and included qualitative and semiquantitative light microscopy. LANR performed with a protein solder results in a good early peripheral nerve regeneration, with an optimal alignment of nerve fibers and minimal connective tissue proliferation at the repair site. All three suture materials produced a foreign body reaction; the least severe was with polyglycolic acid sutures. CMSR resulted in more pronounced foreign-body granulomas at the repair site, with more connective-tissue proliferation and axonal misalignment. Furthermore, axonal regeneration in the distal nerve segment was better in the laser groups. Based on these results, CO(2) laser-assisted nerve repair with soldering in combination with absorbable sutures has the potential of allowing healing to occur with the least foreign-body reaction at the repair site. Further experiments using this combination are in progress. PMID- 12740883 TI - Synergistic effect of vincristine with tacrolimus or sirolimus in prevention of acute heart allograft rejection in the rat. AB - The application of multiple immunosuppressive therapy for organ transplantation could enhance therapeutic efficacy, while minimizing the toxicity of individual drugs used in the regimen. In this study, the effect of the combined therapy of vincristine (VCR) with tacrolimus (FK506) or sirolimus (rapamycin, RAPA) was tested in prevention of acute heart allograft rejection in the rat. A Brown Norway (BN, RT 1(n)) to Lewis (LEW, RT 1(1)) rat combination was used in a heart allografting model. VCR was administered intraperitoneally once daily, while FK506 and RAPA were given by gavage once daily for 14 days after transplantation. There were dose-related prolongations of mean survival time (MST) to monotherapy of VCR, FK506, or RAPA. The MST in combination therapy indicated that a synergistic interaction was produced when compared with the respective monotherapies: VCR 0.05 mg/kg/day + FK506 0.5 mg/kg/day (16.00 +/- 1.79 days, P = 0.001; combination index (CI) = 0.557); VCR 0.05 mg/kg/day + FK506 1.0 mg/kg/day (29.00 +/- 10.54 days, P = 0.001; CI = 0.598); VCR 0.05 mg/kg/day + RAPA 0.2 mg/kg/day (17.33 +/- 1.97 days, P = 0.001; CI = 0.500); and VCR 0.05 mg/kg/day + RAPA 0.4 mg/kg/day (21.17 +/- 3.19 days, P = 0.001; CI = 0.838). Combination therapy of VCR and FK506 or RAPA produced synergistic effects in prevention of acute heart allograft rejection in the rat. PMID- 12740884 TI - Biodegradable, thermosensitive implant for approximating cylindrical structures: a preliminary study. AB - A new invention, i.e., a biodegradable, thermosensitive hybrid gel composite in the form of a thin sheet, was used as an implant inside the rat and guinea pig to test its tissue reactions, degradation, and function as a bridging agent. Tissue tractions in subcutaneous tissue, in muscle, around a peripheral nerve, and around an artery were mild. Degradation made rapid progress, starting on the third day and completed in 2-3 weeks. The hybrid gel, when wrapped around the cut ends of a peripheral nerve or an artery, functioned well as a bridging tube. In the case of the cut nerve, regenerating fascicles crept through the tube which protected them from fibrosis. In the case of the cut artery, patency of flow was maintained, and arterial wall healing was complete in 2-3 weeks. The hybrid gel composite contracted at body temperature, thus holding well to body tissues. Its biodegradable and inert nature offers the potential for future use as a tissue wrap and bridging agent. PMID- 12740885 TI - Retrograde arterialized venous flap: an experimental study. AB - An experimental model was established to study circulation in retrograde arterialized venous flaps (RAVF). Venous flaps measuring 7 x 4 cm with a matching venous system were harvested from both forearms of 10 fresh human cadavers. In each trial, both flaps were simultaneously perfused with heparinized human blood driven by a pulsatile circulation model. In each trial there was one flap with retrograde perfusion, and one flap with antegrade perfusion. Clinical assessment, measurement of outflow, and angiographic examination with digitally assisted assessment after 3 h of perfusion showed better results for retrograde perfusion in 8 of the 10 trials. This study indicates that blood circulation in the periphery of arterialized venous flaps can be enhanced by retrograde arterialization. PMID- 12740886 TI - Rat model of laryngeal transplantation with normal circulation maintained by combination with the tongue. AB - Recent research on laryngeal transplantation used a rat model with physiologically abnormal circulation, because the common carotid artery was used for venous drainage. Since a model with normal circulation is better for accurate assessment, we developed a vascularized laryngeal graft, using the common carotid artery as the arterial supply and the external jugular vein as the venous drainage. The transplant included other tributaries from the tongue and the pharynx by combination with the tongue, because the tributaries from the larynx were very small. Ten transplants were performed. The transplanted organs were examined daily. Seven days after transplantation, the composite grafts were evaluated histologically. The mortality rate was 10%. All composite grafts were fully viable, and their structure was maintained in the remaining nine rats. Our model is appropriate for experiments on laryngeal transplantation. PMID- 12740887 TI - Impact of poor microsurgical suture technique on tissue perfusion in a rat model. AB - This study was designed to evaluate the effect of poor suture technique in microsurgical small-vessel anastomosis on tissue microcirculation. Eighteen male Sprague-Dawley rats were divided into three groups of six animals each: sham control group, regular suture group (control), and inverting suture group. In the regular suture group, a standard single-stitch suture technique was used for microsurgical reanastomosis of the transected common iliac artery. In the inverting suture group, a suture technique was applied inverting the vessel wall, thus simulating poor suture technique. After 24 h, intravital microcirculatory measurements were obtained in the cremaster muscle flap. The inverting suture caused a significant drop in capillary perfusion from 6.40 (sham control) and 5.65 (regular suture) to 1.81 capillaries per visual field (P < 0.005). A poor microsurgical anastomosis may result in a significant reduction of peripheral tissue perfusion, although blood flow through the main feeding vessel is maintained. PMID- 12740888 TI - Microsurgery simulators in virtual reality: review. AB - Surgical training is undergoing a rapid transformation, which has been influenced by advances in computer modeling. Increased pressure to reduce the use of animals in technical training has led to a new approach in teaching microsurgery. This new technology may prove to be a cost-effective, portable, and nonhazardous way forward in microsurgical training. This paper reviews the current state of available technological models used in microsurgical training. In doing so, we review articles from the latest journals and authenticated Internet websites to compare and contrast these various methods. Finally, we look at the specific technique that has potential impact on the future modeling of microsurgical techniques. PMID- 12740889 TI - Development and validation of new model for microvascular transplantation of epiphyseal plate allografts with minimal adjoining epiphyseal and metaphyseal bone. AB - A model for the free allograft microvascular transplantation of rabbit proximal tibial epiphyseal plate allografts was developed, validated, and tested in an in vivo animal model. Transplants contained the minimum amount of adjacent epiphyseal and metaphyseal bone compatible with preservation of the epiphyseal plate vascular supply, as determined by corrosion casting. Perfusion to this graft was evaluated quantitatively using radioactive microspheres, and qualitatively using India-ink injection. Female New Zealand White rabbits at 12 weeks of age were utilized. Vascularized transplantation of epiphyseal plate allografts was performed either into a defect of matched size in the iliac crest or into a soft-tissue pocket without bone contact. Cyclosporine A immunosuppression (CSA) was administered daily for 6 weeks. Two control groups underwent identical surgical procedures, but had no postoperative immunosuppression. Epiphyseal plates both with and without bone contact, in rabbits immunosuppressed postoperatively with CSA, demonstrated longitudinal growth and preserved viability as determined by positive bromodeoxyuridine uptake. Control epiphyseal plates transferred without postoperative immunosuppression were uniformly nonviable. This new model has value as a basis for further studies into the clinical applicability of isolated epiphyseal-plate transplants. PMID- 12740890 TI - New model for simultaneous heart and kidney transplantation in mice. AB - In clinical settings, combined heart and kidney transplantation results in a lower incidence of cardiac graft and renal graft rejection as compared to isolated heart and kidney transplantation. To study the phenomenon in an experimental setting, we developed a model of combined heart and kidney transplantation in mice. According to our technique, we kept the patch of the infrarenal aorta and inferior vena cava (IVC) as long as possible during the donor's kidney explantation, and then, after finishing the kidney implantation with conventional techniques, we anastomosed the innominate artery and pulmonary artery of the heart graft to the patch of the infrarenal aorta and IVC of the renal graft, respectively, instead of the recipient's aorta and IVC. With the use of our method, combined heart and kidney transplantation in mice can be performed to get enough suture room for the second graft, to avoid prolonging the occlusion time of the recipient's circulation, and to profoundly decrease postoperative complications such as paraplegia and mortality. Of the 14 recipients with combined heart-kidney isografting, 10 have been successful (71.4%), surviving over 100 days with normal function of both grafts, and with lack of change in histological appearance. This suggests that the technique is feasible and reliable. PMID- 12740891 TI - Statistical estimators of frontal sinus cross section ontogeny from very noisy data. AB - Cross-sectional areas of human frontal sinuses in the occipitofrontal projection are a good surrogate for frontal sinus volumes. This study looks at these areas in a dataset of some 200 children and 100 adults of both sexes. As measured by planimetry of roentgenograms, the areas are extremely variable ("noisy" in a statistical sense). In fact, they appear to be distributed log-normally with quite high variance. The mean of the distribution is evidently a function of age and the variances differ by sex. After logarithmic transformation, the data are adequately fitted by one sigmoid curve for each sex. Our discussion highlights implications of this finding for the biological aspects of frontal sinuses and methodological issues in ontogenetic analysis of data so noisy. PMID- 12740892 TI - Sperm ultrastructure in the nudibranch genus Halgerda with reference to other Discodorididae and to Chromodorididae (Mollusca: Opisthobranchia). AB - Comparative sperm ultrastructure within the molluscan nudibranch genus Halgerda (Discodorididae) was examined for the first time using transmission electron microscopy (TEM), based on 17 of the 35 known species. In addition, observations on two other discodorids are made to facilitate outgroup comparison with Halgerda, including one species of Discodoris (D. boholiensis) and Asteronotus cespitosus (currently accepted as the closest sister taxon to Halgerda). Comparison was also made with some genera of the Chromodorididae in view of sperm similarities. Spermatozoa of all species examined were of the complex, helical, elongate ( approximately 300-400 micro m) type characteristic of most heterobranch gastropods. These cells exhibit the following discrete regions (in anteroposterior sequence) : an acrosomal complex (composed of a rounded, membrane bound vesicle and a column-like pedestal); a solid, helical nucleus; an elongate, helical midpiece (composed of an axoneme and associated nine coarse fibers, an enveloping mitochondrial derivative of matrix, and paracrystalline materials and glycogen helix); an annular complex; and a short glycogen piece. Of these regions, the midpiece is by far the longest, occupying over 90% of the total sperm length. Comparison with other members of the radula-bearing cryptobranch dorids reveals several sperm similarities to other genera in the clade, particularly those of other Discodorididae and also with the Chromodorididae. Comparison with previously studied genera reveals noteworthy sperm differences within the Discodorididae. The most notable differences are the internal structure of the acrosomal pedestal (long and homogeneous in Halgerda, Discodoris; short and homogeneous in Asteronotus; long and finely striated in Rostanga; oblong with angular electron-lucent striations in Jorunna) and the internal structure of the glycogen piece. The pronounced helical keels of most Halgerda and Discodoris nuclei contrast with the weakly helical nucleus of Asteronotus. Sperm features alone do not provide a means of defining the genus Halgerda or the family Discodorididae nor do they support the monophyletic status of the caryophyllidia-bearing dorids. Important sperm characters such as the acrosome, nucleus, and midpiece can often still be determined from specimens that have been initially fixed in formalin, then stored in ethanol for extended periods of time (i.e., museum material). Of all sperm features, the mitochondrial derivative of the midpiece is the most resistant to long-term fixation : the survival of acrosomal, nuclear, and axonemal components is variable, presumably a factor of prefixation autolysis, varied primary fixation times and temperatures, formalin quality, and duration of alcohol storage. PMID- 12740893 TI - Winged presoldiers induced by a juvenile hormone analog in Zootermopsis nevadensis: implications for plasticity and evolution of caste differentiation in termites. AB - To elucidate the switching mechanism of caste differentiation in termites and to examine the possible induction of soldier-reproductive intercastes experimentally, we investigated the effects of juvenile hormone on the morphologies of soldier caste by applying a juvenile hormone analog (JHA) to nymphs of the damp-wood termite Zootermopsis nevadensis (Isoptera : Termopsidae). JHA treatment for about 2 weeks induced a variety of intermediate castes, showing both alate and soldier morphological features. The principal component analysis (PCA) of those morphological characters showed that those intercastes were a deviation from the developmental line into alates to soldier differentiation, which is known to be triggered by juvenile hormone. Detailed morphological examination of the compound eyes, wing joint, and mandibles showed that those intercastes expressed soldier features, although they had started to develop alate characteristics. The morphology of the resultant intercastes seemed to be determined by the nymphal stage, at which JHA treatment was applied. The induced intercastes with exaggerated soldier-specific characteristics (e.g., mandibles) repressed alate-specific characteristics (e.g., wings), namely, the alate and soldier morphological characteristics in induced intercastes show opposite responses against the application of JHA. On the other hand, ovarian development was not suppressed by the JHA application, even in the soldier-like individuals. Naturally differentiated presoldiers also possessed developed ovarioles, although ovaries of mature soldiers were degenerated. Our results suggest that the juvenile hormone plays complicated roles in the expression of caste morphologies and ovarian development in termites. PMID- 12740894 TI - Biomechanics of the rostrum and the role of facial sutures. AB - The rostrum is a large diameter, thin-walled tubular structure that receives loads from the teeth. The rostrum can be conceptualized both as a rigid structure and as an assemblage of several bones that interface at sutures. Using miniature pigs, we measured in vivo strains in rostral bones and sutures to gain a better understanding of how the rostrum behaves biomechanically. Strains in the premaxillary and nasal bones were low but the adjacent maxillary-premaxillary, internasal, and intermaxillary suture strains were larger by an order of magnitude. While this finding emphasizes the composite nature of the rostrum, we also found evidence in the maxillary and nasal bones for rigid structural behavior. Namely, maxillary strain is consistent with a short beam model under shear deformation from molar loading. Strain in the nasal bones is only partially supported by a long beam model; rather, a complex pattern of dorsal bending of the rostrum from incisor contact and lateral compression is suggested. Torsion of the maxilla is ruled out due to the bilateral occlusion of pigs and the similar working and balancing side strains, although it may be important in mammals with a unilateral bite. Torsional loading does appear important in the premaxillae, which demonstrate working and balancing side changes in strain orientation. These differences are attributed to asymmetrical incisor contact occurring at the end of the power stroke. PMID- 12740895 TI - Structure of spermatodoses in shield-back bushcrickets (Tettigoniidae, Tettigoniinae). AB - Many aspects of the reproductive anatomy and physiology of tettigoniids have been studied extensively. These include the large, externally visible spermatophores and the bundles of sperm, known as spermatodesms. However, spermatodoses, spermatophore-like structures found within the spermatheca, seem to have been almost completely overlooked: their structure has not been described since 1913 and they have subsequently received only passing mention in the literature. Each time the female mates, a separate spermatodose is formed. Here I use photographs, from light-microscopy, of whole and sectioned spermatodoses to describe the external and internal structure of spermatodoses of nine different genera within the subfamily Tettigoniinae. The structure of the spermatodoses is very similar for the different genera. Each spermatodose is pear- or onion-shaped and consists of a thin outer layer, enclosing a thick, gelatinous inner layer. A large sperm mass occupies the bulbous end of the spermatodose, while a thin sperm-tube leads from the sperm mass, along the center of the elongated neck of the spermatodose, and appears to exit at the pointed-tip of the spermatodose. Feather-like bundles of sperm (spermatodesms) were clearly visible within the sperm mass and also appeared to be present within the sperm-tube. The wall of the sperm tube appeared to be composed of material similar to that of the outer layer of the spermatodose. Within the spermatheca, spermatodoses appeared to be stratified in that only one of them ever occupied the position nearest to the spermathecal duct. The possible function of spermatodoses is discussed: it is proposed that they have evolved as a result of sexual conflict and function to protect the sperm from being destroyed by the female while they are in storage. PMID- 12740896 TI - A desert octodontid rodent, Tympanoctomys barrerae, uses modified hairs for stripping epidermal tissue from leaves of halophytic plants. AB - Desert rodents that consume halophytic plants must have adaptations for coping with the high salt content of the leaves. A kidney capable of excreting very concentrated urine is one method. Another is removal of the hypersaline epidermis by means of chisel-like incisors prior to ingestion of the leaves. Tympanoctomys barrerae has evolved a unique refinement of the latter adaptation. It possesses two bundles of stiffened hairs on either side of the palate just caudal to the incisors. The bundles vibrate against the lower incisors, removing the epidermis from the leaves. The efficiency of the operation is significantly greater than with the use of incisors alone. Such a device has not been described in any other mammal. The facial muscles associated with the lips, the cheek vibrissae, and the oral cavity are described in T. barrerae and the nonhalophilic octodontid Octomys mimax. M. buccinatorius pars intermaxillaris is the only muscle in direct contact with the bristle bundles. Other anatomical features found in T. barrerae that may be associated with this feeding device are: 1) a much enlarged and mobile lower labial pad operated by Mm. buccinatorius pars orbicularis oris, pars longitudinalis profunda, and mandibularis cranialis profunda; 2) two oral glands not described in other rodents; and 3) a shortened tongue. Although, taken as a whole, this epidermal stripping device is unique to T. barrerae, most of its features have evolved by modification of structures present in the facial region of more generalized rodents. PMID- 12740897 TI - Morphology and histochemistry of the peripheral olfactory organ in the round goby, Neogobius melanostomus (Teleostei: Gobiidae). AB - This first comprehensive study of the peripheral olfactory organ from a representative of the large and economically important order of teleost fishes, the Perciformes, shows a compact structure with olfactory sensory neurons distributed widely throughout the olfactory chamber. The spatial organization of the nasal cavity in the bottom-dwelling round goby (Gobiidae, Neogobius melanostomus) was examined using impression material injection, immunocytochemistry, and transmission electron microscopy. The olfactory chamber contains a single olfactory lamella; prominent dorsocaudal lachrymal and ethmoidal accessory nasal sacs are situated ventrocaudal to the chamber. The location of the olfactory mucosa within the olfactory chamber is novel for teleost fish, as it extends beyond the ventral surface to the lateral and dorsal regions. Microvillar olfactory sensory neurons and ciliated olfactory sensory neurons were identified by transmission electron microscopy and the spatial distribution of these two cell types was assessed through immunocytochemistry against olfactory receptor coupled G-proteins. Both G(alphaolf)-immunoreactive ciliated olfactory sensory neurons and the G(alphao)-immunoreactive microvillar form were located throughout the olfactory epithelium. Ciliated crypt cells were G(alphao) immunoreactive and were found throughout the olfactory epithelium of some specimens. The widespread occurrence of olfactory sensory neurons in the olfactory chamber supports the idea that olfactory signaling is important to the survival of the round goby. The prominence of the lachrymal and ethmoidal accessory nasal sacs indicates the capacity to regulate the flow of odorant molecules over the sensory surface of the olfactory sensory neurons, possibly through a pump-like mechanism driven by opercular activity associated with gill ventilation. PMID- 12740898 TI - Pericardium of the frog, Rana esculenta, is morphologically designed as a lymphatic space. AB - The importance of the pericardium and the pericardial fluid (PF) in the control of cardiac function has emerged over the past few years. Despite the acknowledgment that amphibians are exposed to both dehydration and excessive water accumulation, nothing is known about their pericardial structure and the morphological basis of the PF formation. We have studied the parietal pericardium (PP) morphology in Rana esculenta by electron microscopy. SEM images of the inner surface, which lines the pericardial cavity, revealed the presence of large vesicles and many small circular openings. TEM observations showed that the PP is made up of an inner mesothelial lining, often constituted by two layers of very flat cells lying on a basal membrane and of regularly oriented collagen bundles. The PP outer surface is lined by a layer of flat cells, without a basal membrane. The mesothelial cells had overlapping boundaries with complex intercellular connections and a rich pool of caveolae opened in the direction of both the pericardial cavity and intercellular spaces. These cells indicate an intense intracellular and/or intercellular transfer of fluids and substances. The intraperitoneal injection of the idromineral hormone, Val(5)-ANG II, induced PP modifications, particularly evident at the level of the structures involved in the transmesothelial traffic. These lymphatic-like traits suggest that the frog PP represents a large lymphatic sac, subject to paracrine-endocrine remodeling, which can actively adjust the PF, influencing the composition and volume of the myocardial interstitial fluid. PMID- 12740900 TI - Interspecific variations in the morphology and ultrastructure of the rhabdoms of oplophorid shrimps. AB - Interspecific variations in rhabdom structure between various oplophorid shrimps are described and the differences are related to the light environment at different depths within the mesopelagic zone. The ultrastructure of the distal rhabdom in these species is described for the first time. Quantitative measurements show that the proportion of the rhabdom layer occupied by the distal rhabdom varies from 3.5-25% in the dorsoventral plane of the eye of Systellaspis debilis. The distal rhabdom occupies less than 1% of the rhabdoms in the eye of Acanthephyra pelagica, where it can only be seen by using the electron microscope. It is suggested that the rhabdoms of those species that remain within the photic zone (such as S. debilis) are adapted to maximize contrast, whereas in those whose depth ranges extend into the aphotic zone (such as A. pelagica) they are adapted for maximum sensitivity. PMID- 12740899 TI - Spatial expression patterns of skin-type antifreeze protein in winter flounder (Pseudopleuronectes americanus) epidermis following metamorphosis. AB - Two isotypes of Type I antifreeze protein (AFP), the liver-type and the skin type, have been described from adult winter flounder (Pseudopleuronectes americanus). Although the liver-type AFP has been well studied, the skin-type has just begun to be characterized. It appears to have a wide tissue distribution, be expressed constitutively, and the absence of a signal sequence suggests it is active intracellularly. The current study was designed to examine the onset of skin-type AFP expression during the thickening of the epidermis at metamorphosis from both the nucleic acid and protein levels. The epidermis appeared as a thin layer overlying a thickened dermis at metamorphosis and showed a gradual increase in thickness through the first fall and winter. The onset of skin-type antifreeze expression occurred in conjunction with this epidermal thickening. In situ hybridization and immunohistochemistry showed a distribution of mRNA and skin type AFP specific for the epidermis and epidermal pavement cells. The AFP immunoproduct showed a distribution intimate with the pavement cell membrane and through the interstitial spaces. This distribution suggests that the AFP may be important in slowing ice crystal formation in these interstitial regions and thus reducing cellular damage due to osmotic imbalance. PMID- 12740901 TI - Functional morphology of extreme jaw protrusion in Neotropical cichlids. AB - The New World cichlids Petenia splendida and Caquetaia spp. possess extraordinarily protrusible jaws. We investigated the feeding behavior of extreme (here defined as greater than 30% head length) and modest jaw-protruding Neotropical cichlids by comparing feeding kinematics, cranial morphology, and feeding performance. Digital high-speed video (500 fps) of P. splendida, C. spectabile, and Astronotus ocellatus feeding on live guppy prey was analyzed to generate kinematic and performance variables. All three cichlid taxa utilized cranial elevation, lower jaw depression, and rotation of the suspensorium to protrude the jaws during feeding experiments. Extreme anterior jaw protrusion in P. splendida and C. spectabile resulted from augmented lower jaw depression and anterior rotation of the suspensorium. Morphological comparisons among eight cichlid species revealed novel anterior and posterior points of flexion within the suspensorium of P. splendida and Caquetaia spp. The combination of anterior and posterior loosening within the suspensorium in P. splendida and Caquetaia spp. permitted considerable anterior rotation of the suspensorium and contributed to protrusion of the jaws. Petenia splendida and C. spectabile exhibited greater ram distance and higher ram velocities than did A. ocellatus, resulting primarily from increased jaw protrusion. Petenia splendida and C. spectabile exhibited lower suction feeding performance than A. ocellatus, as indicated by lower suction-induced prey movements and velocities. Thus, extreme jaw protrusion in these cichlids may represent an adaptation for capturing elusive prey by enhancing the ram velocity of the predator but does not enhance suction feeding performance. PMID- 12740902 TI - Involvement of the gonadal germinal epithelium during sex reversal and seasonal testicular cycling in the protogynous swamp eel, Synbranchus marmoratus Bloch 1795 (Teleostei, Synbranchidae). AB - The swamp eel, Synbranchus marmoratus, is a protogynous, diandric species. During sex reversal, the ovarian germinal epithelium, which forms follicles containing an oocyte and encompassing follicle cells during the female portion of the life cycle, produces numerous invaginations, or acini, into the ovarian stroma. Within the acini, the gonia that formerly produced oocytes become spermatogonia, enter meiosis, and produce sperm. The acini are bounded by the basement membrane of the germinal epithelium. Epithelial cells of the female germinal epithelium, which formerly became follicle (granulosa) cells, now become Sertoli cells in the developing testis. Subsequently, lobules and testicular ducts form. The swamp eel testis has a lobular germinal compartment in both primary and secondary males, although the germinal compartment in testes of secondary males resides within the former ovarian lamellae. The germinal compartment, supported by a basement membrane, is composed of Sertoli and germ cells that give rise to sperm. Histological and immunohistochemical techniques were used to describe the five reproductive classes that were observed to occur during the annual reproductive cycle: regressed, early maturation, mid-maturation, late maturation, and regression. These classes are differentiated by the presence of continuous or discontinuous germinal epithelia and by the types of germ cells present. Synbranchus marmoratus has a permanent germinal epithelium. Differences between the germinal compartment of the testes of primary and secondary males were not observed. PMID- 12740903 TI - Telomerase activity in high-grade cervical lesions is associated with allelic imbalance at 6Q14-22. AB - Our study attempts to establish the relationship between telomerase activity and allelic imbalance (AI) on chromosomes 3p and 6 in high-risk HPV-containing cervical lesions. These chromosomes were implicated previously in telomerase regulation in HPV containing immortalized cells and cervical cancer cells. Allelotyping and telomerase analysis were carried out on 28 high-grade cervical lesions (CIN III: n = 20; cervical carcinomas: n = 8), using 23 microsatellite markers on 3p, 6p and 6q. Clear telomerase activity was found in 17 of 28 lesions (61%). Allelic imbalance frequency at 6q14-22 was significantly higher in lesions with detectable telomerase activity, compared to lesions without telomerase activity (p = 0.02). No association was found between telomerase activity and AI at any of the remaining regions studied on 3p and chromosome 6. In addition, in telomerase positive passages of the HPV 16 immortalized cell line FK16A, shown recently to be responsive to chromosome 6 mediated telomerase repression, AI was found in the overlapping region of 6q14-27. These data suggest that 6q14-22 may contain 1 or more genes involved in telomerase deregulation and immortalization during cervical carcinogenesis. PMID- 12740904 TI - Activation of 3-nitrobenzanthrone and its metabolites by human acetyltransferases, sulfotransferases and cytochrome P450 expressed in Chinese hamster V79 cells. AB - 3-nitrobenzanthrone (3-NBA) is a potent mutagen and suspected human carcinogen identified in diesel exhaust and ambient air pollution. 3-aminobenzanthrone (3 ABA), 3-acetylaminobenzanthrone (3-Ac-ABA) and N-acetyl-N-hydroxy-3 aminobenzanthrone (N-Ac-N-OH-ABA) have been identified as 3-NBA metabolites. Recently we found that 3-NBA and its metabolites (3-ABA, 3-Ac-ABA and N-Ac-N-OH ABA) form the same DNA adducts in vivo in rats. In order to investigate whether human cytochrome P450 (CYP) enzymes (i.e., CYP1A2), human N,O-acetyltransferases (NATs) and sulfotransferases (SULTs) contribute to the metabolic activation of 3 NBA and its metabolites, we developed a panel of Chinese hamster V79MZ-h1A2 derived cell lines expressing human CYP1A2 in conjunction with human NAT1, NAT2, SULT1A1 or SULT1A2, respectively. Cells were treated with 0.01, 0.1 or 1 microM 3 NBA, or its metabolites (3-ABA, 3-Ac-ABA and N-Ac-N-OH-ABA). Using both enrichment versions of the (32)P-postlabeling assay, nuclease P1 digestion and butanol extraction, essentially 4 major and 2 minor DNA adducts were detected in the appropriate cell lines with all 4 compounds. The major ones were identical to those detected in rat tissue; the adducts lack an N-acetyl group. Human CYP1A2 was required for the metabolic activation of 3-ABA and 3-Ac-ABA (probably via N oxidation) and enhanced the activity of 3-NBA (probably via nitroreduction). The lack of acetylated adducts suggests N-deacetylation of 3-Ac-ABA and N-Ac-N-OH ABA. Thus, N-hydroxy-3-aminobenzanthrone (N-OH-ABA) appears to be a common intermediate for the formation of the electrophilic arylnitrenium ions capable of reacting with DNA. Human NAT1 and NAT2 as well as human SULT1A1 and SULT1A2 strongly contributed to the high genotoxicity of 3-NBA and its metabolites. Moreover, N,O-acetyltransfer reactions catalyzed by human NATs leading to the corresponding N-acetoxyester may be important in the bioactivation of N-Ac-N-OH ABA. As human exposure to 3-NBA is likely to occur primarily via the respiratory tract, expression of CYPs, NATs and SULTs in respiratory tissues may contribute significantly and specifically to the metabolic activation of 3-NBA and its metabolites. Consequently, polymorphisms in these genes could be important determinants of lung cancer risk from 3-NBA. PMID- 12740905 TI - Carboplatin induces Fas (APO-1/CD95)-dependent apoptosis of human tongue carcinoma cells: sensitization for apoptosis by upregulation of FADD expression. AB - We examined the apoptosis of tongue carcinoma cells and the effects of anticancer drugs to identify the molecules that mediate apoptotic cascade in the malignancy. Carboplatin (CBDCA) induced apoptosis of SCC-9 and SCC-25, human well differentiated tongue squamous carcinoma cell lines. Neutralizing anti-Fas (APO 1/CD95) and anti-Fas ligand (FasL) antibodies obliterated the CBDCA-induced cell death. In the absence of CBDCA, cytotoxic anti-Fas antibody, which binds to and activates Fas at the cell surface, failed to induce apoptosis. However, in the presence of CBDCA, the cytotoxic antibody markedly enhanced the apoptosis in a dose-dependent manner. Western blotting and reverse-transcription (RT) PCR revealed that there were no alterations in Fas or FasL expression upon CBDCA treatment. SCC-25 induced apoptosis of Jurkat cells, Fas-sensitive T-lymphatic leukemia cell line, and the apoptosis was inhibited by neutralizing anti-Fas or anti-FasL antibody. These results indicate that the tongue carcinoma cells express nonfunctional Fas and functional FasL, which by themselves fail to induce apoptosis. The expression of FADD in the tongue carcinoma cells was very low and was largely enhanced by CBDCA treatment. Suppression of FADD expression using the specific antisense oligonucleotide resulted in a failure of CBDCA induction of cell death. These results indicate that a deficiency of FADD is involved in the insensitivity of tongue carcinoma cells for Fas activation, and that CBDCA treatment switches nonfunctional Fas to functional Fas by upregulation of FADD expression, resulting in activation of a Fas-sensitive pathway leading to apoptosis. PMID- 12740906 TI - Involvement of PKC betaII in anti-proliferating action of a new antitumor compound gnidimacrin. AB - Daphnane-type diterpene gnidimacrin (NSC 252940) shows significant antitumor activity against murine tumors and human tumor cell lines. This compound binds to and directly activates protein kinase C (PKC), arresting the cell cycle at the G(1) phase through inhibition of cdk2 activity in human K562 leukemia cells. In our study, we examined whether cellular PKC is involved in the antiproliferating effect of gnidimacrin. In a 24-hr exposure of K562 cells to high concentrations of bryostatin 1 (0.11-3.3 microM), both expression of PKC alpha and PKC betaII was downregulated, and thereafter these cells became resistant to gnidimacrin in response to the degree of PKC downregulation. In addition, PKC alpha and PKC betaII genes were transfected to gnidimacrin-resistant human hepatoma HLE cells that demonstrated positive expression of PKC alpha and negative expression of PKC betaII. PKC betaII gene-transfected cells became sensitive to gnidimacrin in relation to the degree of PKC betaII expression. The most sensitive clone to show 0.001 microg/mL (1.2 nM) as IC(50) in a continuous 4-day exposure was obtained. While PKC alpha gene-transfected cells exhibited an increase in PKC alpha expression and became sensitive to gnidimacrin, sensitivity was one-hundredth of that in PKC betaIotaIota gene-transfected cells. These results suggest that PKC, in particular PKC betaIotaIota, is necessary in the antitumor effect of gnidimacrin. PMID- 12740907 TI - Antiestrogens are pro-apoptotic in normal human breast epithelial cells. AB - Estrogens promote cell proliferation in normal and transformed mammary epithelial cells by inducing expression of hormone-responsive genes involved in the cell cycle. The action of antiestrogens is therefore central in regard to their potent inhibitory effects on estrogen-induced cell growth. We used normal human epithelial breast cells from primary cultures (HBE cells) to study hormonal (estrogen and antiestrogen) regulation on 3 key proteins involved in the apoptotic process: Bcl-2, p53 and caspase-3. The mammary adenocarcinoma cell line, MCF-7, was also used to study the molecular regulation of Bcl-2. In both HBE and MCF-7 cells, we found that estradiol (E2) induced an increase in Bcl-2 mRNA levels. This effect was counteracted in the presence of a pure antiestrogen, ICI 182780 (ICI). Alone, ICI did not modify either the Bcl-2 protein or mRNA levels in HBE cells, whereas in MCF-7, a strong downregulation of Bcl-2 mRNA was observed. In parallel, in HBE cells, we observed that E2 caused a decrease in p53 and caspase-3 protein levels, whereas ICI alone increased p53 and caspase-3 protein levels. The ICI effects on p53 and caspase-3 were partially counteracted by E2. Under the same experimental conditions, ICI exerts a potent pro-apoptotic effect, which was not counteracted by E2. In contrast, 4-hydroxytamoxifen was slightly weaker as a pro-apoptotic agent in HBE cells and its effects were reversed by E2. We demonstrate that in HBE cells, ICI reverses the anti-apoptotic action of E2 and alone acts as a highly potent pro-apoptotic molecule. These results provide new insight into treatment for breast cancer prevention. PMID- 12740908 TI - Interspecies contamination of the KM3 cell line: implications for CD63 function in melanoma metastasis. AB - CD63 is a member of the tetraspanin superfamily of membrane glycoproteins that has been hypothesised to provide a structural network in the organisation of large multimolecular microdomains at cell membranes. Detailed analyses of the role of CD63 in these complexes through mutagenic studies have been limited, however, by the ubiquitous cellular expression of CD63 in vivo and in vitro. In an attempt to define CD63-null cell lines, we have analysed the expression of CD63 and other tetraspanins on a panel of human cancer cell lines. Similar expression patterns were seen between cell lines from melanomas, breast cancers and prostate cancers. The melanoma cell line KM3, however, described previously as a CD63-null human cell line, was found to express none of the 7 human tetraspanins tested. KM3 was identified definitively as a rat cell line by analysis of karyotype and antigen expression. Notably, KM3 was found to express the rat homologue of CD63. Conclusions concerning the function of human CD63 drawn from studies using KM3 cells therefore require re-evaluation as does the frequently cited hypothesis that CD63 expression is linked to melanoma progression. As KM3 is the only cell line thus far identified as CD63 negative, these results highlight the necessity for the production of a CD63 null system. PMID- 12740909 TI - Development of resistance to a trinuclear platinum complex in ovarian carcinoma cells. AB - BBR3464 is a trinuclear platinum complex that exhibits a potent cytotoxicity and efficacy against cisplatin-resistant tumors. To better understand the determinants of cellular resistance to BBR3464, we selected a resistant ovarian carcinoma cell line after exposure to the complex. The resistant cells (A2780/BBR3464) exhibited a high level of resistance to the selecting agent, but a marginal cross-resistance to cisplatin. Although cellular accumulation of BBR3464 was similar in parental and in resistant cells, DNA platination was decreased in A2780/BBR3464 cells, suggesting a reduced drug accessibility to DNA. This behavior reflected a partial drug inactivation at cytoplasmic level, as a consequence of increased levels of nucleophilic molecules including metallothioneins and human neurofilament low, but not glutathione. A2780/BBR3464 cells also exhibited a reduced susceptibility to apoptosis, which was consistent with reduced expression of Bax, and an alteration of DNA mismatch repair system, as reflected by lack of expression of MLH1 and PMS2, which could impair the recognition/repair of DNA lesions. Whereas both platinum drugs induced G2/M arrest in the parental cells, BBR3464, but not cisplatin, caused a late G1 arrest of resistant cells. Cisplatin induced an appreciable increase of p21(WAF1) levels in both models, in contrast to BBR3464 that produced a substantial upregulation of p21(WAF1) only in parental cells. An inverse relationship with p21(WAF1) modulation was found for CHK1 in parental cells treated with both agents and in resistant cells treated with cisplatin. This pattern of response is consistent with a regulatory loop involving p53 and p21(WAF1) at G2 checkpoint. In contrast, no modulation of CHK1 was found in A2780/BBR3464 treated with the triplatinum compound. These findings, indicating a different activation of regulatory pathways at DNA damage checkpoints in response to cisplatin and BBR3464, support an altered ability of resistant cells to recognize or tolerate sublethal lesions induced by BBR3464. PMID- 12740910 TI - KLF6, a putative tumor suppressor gene, is mutated in astrocytic gliomas. AB - Gliomas are the most common tumors of the central nervous system and have a grave prognosis. Deletion of chromosome 10p15 is one of the most common chromosomal alterations in gliomas. Recently, a candidate tumor suppressor gene, KLF6, which is mapped to chromosome 10p, was found to be frequently mutated in prostate cancer. KLF6 is a zinc finger transcription factor and transactivates p21/WAF1/CIP expression. To elucidate the role of genetic alterations of KLF6 in gliomas, we analyzed the 4 exons of the gene by direct DNA sequencing in 155 gliomas. Of these, mutations of KLF6 were found in 9 of 76 (11.8%) glioblastomas multiforme, 2 of 28 (7.1%) anaplastic astrocytomas, 2 of 36 (5.5%) low-grade diffuse astrocytomas and in none of the 15 oligodendrogliomas. All 13 mutations were located in the transactivation domain and most of them affected either serine residues or codons next to serine residues. Of the 13 cases with KLF6 mutation, loss of heterozygosity (LOH) at the KLF6 locus was inferred from the LOH displayed by the flanking microsatellite markers in 11 cases. We conclude that mutations of the KLF6 gene play a role in the pathogenesis of astrocytic gliomas. PMID- 12740911 TI - Oligogenic segregation analysis of hereditary prostate cancer pedigrees: evidence for multiple loci affecting age at onset. AB - Previous studies have suggested strong evidence for a hereditary component to prostate cancer (PC) susceptibility. Here, we analyze 3,796 individuals in 263 PC families recruited as part of the ongoing Prostate Cancer Genetic Research Study (PROGRESS). We use Markov chain Monte Carlo (MCMC) oligogenic segregation analysis to estimate the number of quantitative trait loci (QTLs) and their contribution to the variance in age at onset of hereditary PC (HPC). We estimate 2 covariate effects: diagnosis of PC before and after prostate-specific antigen (PSA) test availability, and presence/absence of at least 1 blood relative with primary neuroepithelial brain cancer (BC). We find evidence that 2 to 3 QTLs contribute to the variance in age at onset of HPC. The 2 QTLs with the largest contribution to the total variance are both effectively dominant loci. We find that the covariate for diagnosis before and after PSA test availability is important. Our findings for the number of QTLs contributing to HPC and the variance contribution of these QTLs will be instructive in mapping and identifying these genes. PMID- 12740912 TI - Identification of drug-regulated genes in osteosarcoma cells. AB - The introduction of systemic chemotherapy improved significantly the prognosis of osteosarcoma. Despite this success, approximately 30-40% of patients will relapse. Cytotoxic drugs have been shown to induce apoptosis in the target cells independent of their primary effects. The underlying molecular mechanisms and the intracellular mediators, however, are still largely unknown. Therefore, the purpose of our study was to identify drug-regulated genes in osteosarcoma cells useful as prognostic factors and for the development of new therapeutic strategies. Using suppressive subtractive hybridization (SSH) the gene expression pattern of untreated Saos-2 cells was compared to cells treated with cisplatin, methotrexate and doxorubicin, respectively. We identified 8 genes that are regulated >2-fold in drug-treated osteosarcoma cell lines. Expression of ferritin light chain, rhoA, inosine monophosphatdgehydrogenase II, ribonucleotide reductase M2, pro2000 and pro1859 were increased after drug treatment, whereas prohibitin and alpha-actinin expressions were significantly downregulated. Differential expression of the identified genes was verified by Northern blot analysis of 3 different osteosarcoma cell lines. In addition, the effects on chemosensitivity of 4 selected genes was analyzed by overexpression of recombinant constructs in Saos-2 cells and subsequent quantification of drug induced apoptosis. Overexpression of prohibitin and rhoA reduced significantly drug sensitivity to approximately 52% and 59% indicating a crucial role in the modulation of drug-induced cell death. PMID- 12740913 TI - EBV-encoded EBNA-5 associates with P14ARF in extranucleolar inclusions and prolongs the survival of P14ARF-expressing cells. AB - Epstein-Barr virus (EBV) carrying lymphoblastoid cells of normal origin express the full program of all 9 virus-encoded, growth transformation associated proteins. They have an intact p53 pathway as a rule. This raises the question of whether any of the viral proteins impair the pathway functionally. Using a yeast 2-hybrid system, we have shown that EBNA-5 but not the other EBNAs interacts with the p14ARF protein, a regulator of the p53 pathway. The interaction was confirmed in vitro using a GST pull-down assay. Moreover, expression of EBNA-5 increased the survival of p14ARF-transfected cells. EBV infection of resting B cells induced the expression of p14ARF mRNA without increased level of the protein. A fraction of the p14ARF localized to the nucleoli but the bulk of the protein accumulated in nuclear but extranucleolar inclusions. Formation of the extranucleolar inclusions led to complete relocalization of EBNA-5 from nucleoplasm to these structures. The inclusions also contained p53 and HDM2, and were surrounded by PML bodies and proteasomes, which suggests that these inclusions could be targets for proteasome dependent protein degradation. PMID- 12740914 TI - Mutational events in LMP1 gene of Epstein-Barr virus in salivary gland lymphoepithelial carcinomas. AB - It is still unknown what kinds of roles Epstein-Barr virus (EBV) infection that are highly specific to salivary gland lymphoepithelial carcinomas (LECs) play in their tumorigenesis. To clarify the significance of EBV in LECs, we paid particular attention to the LMP1 gene, which is responsible for triggering several pathways for activating transcription factors. Sixty-one cases of EBV positive LECs confirmed by PCR and in-situ hybridization were collected from various areas of the world and studied immunohistochemically for latent membrane protein-1. Furthermore, PCR for the LMP1 carboxyl (C)-terminus region was performed, and the PCR products were sequenced for detection of other mutational events. LMP1 gene products were immunohistochemically demonstrated in 51% of the cases, while PCR amplification of the LMP1 gene was successful in 41 cases (67%). Among them, a 30 bp deletion in the C-terminus of the LMP1 gene, which had been shown to be characteristic to EBV in Chinese nasopharyngeal carcinomas, was found in 20 cases (32%). Most of them were from Guangzhou, Chengdu and Taiwan, while most of the cases from Shanghai and other areas exhibited no 30 bp deletion. In addition, several point mutations including codon 338 of LMP1 were commonly shared by the cases with or without the 30 bp deletion. These results indicate that there are 2 major genomic variations of EBV infecting salivary gland LECs. The frequent mutational events in the C-terminus in addition to the 30 bp deletion also seem to be critical for the pathogenesis because such mutational events may possibly promote cellular proliferation. PMID- 12740915 TI - Repression of Smad-dependent transforming growth factor-beta signaling by Epstein Barr virus latent membrane protein 1 through nuclear factor-kappaB. AB - EBV-encoded LMP-1 is absolutely required for EBV transformation of cells. Previous studies showed that LMP-1 is responsible for mediating resistance to the anti-proliferative effects of TGF-beta that characterizes EBV-transformed cells. To clarify the mechanisms of resistance to TGF-beta by LMP-1, we examined the effect of expression of LMP-1 on the activity of TGF-beta-responsive promoters. Interestingly, LMP-1 inhibited TGF-beta-responsive promoters activity despite lack of direct interaction of LMP-1 and Smad proteins, intracellular signaling molecules in the TGF-beta signal transduction pathway. Although TGF-beta treatment increased the expression of p15, TGF-beta-induced gene, this effect was counteracted by expression of LMP-1. The repressive effect was mapped to the NF kappaB activation domains in the cytoplasmic carboxyl terminus of LMP-1. Furthermore, LMP-1-mediated inhibition of TGF-beta-responsive promoter was markedly restored after inhibition of NF-kappaB activity. LMP-1 failed to affect receptor-dependent formation of heteromers containing Smad proteins as well as the DNA-binding activity of Smad proteins. Overexpression of the transcriptional coactivator CBP and p300 abrogated the inhibitory effect of LMP-1 on the TGF-beta responsive promoter. Our results suggest that LMP-1 represses the TGF-beta signaling through the NF-kappaB signaling pathway at transcriptional level by competing for a limited pool of transcriptional coactivators. These results enhance our understanding of the molecular mechanisms of viral pathogenesis in EBV-associated malignancies. PMID- 12740916 TI - Sequence variations in the DNA repair gene XPD and risk of lung cancer in a Chinese population. AB - Variation in DNA repair capacity, which is believed to be largely determined by genetic traits, is linked to risk of certain cancers. The Asp312Asn and Lys751Gln polymorphisms in the xeroderma pigmentosum complementary group D (XPD) gene may alter DNA repair capacity. We thus examined the hypothesis that these 2 XPD polymorphisms are associated with risk of lung cancer via a large hospital-based, case-control study among Chinese. The study subjects consisted of 1,006 patients with primary lung cancer and 1,020 age- and sex-matched population controls. XPD genotypes were determined using PCR-RFLP techniques, and the associations between genotypes and risk of lung cancer were estimated by odds ratios (ORs) and their 95% confidence intervals (CIs) calculated by unconditional logistic regression. Subjects homozygous for the 312Asn/Asn genotype had an increased risk of lung cancer (adjusted OR = 10.33, 95% CI = 1.29-82.50) compared with subjects homozygous for the 312Asp/Asp genotype. The 751Gln/Gln genotype was also associated with increased risk for the cancer compared with the 751Lys/Lys genotype (adjusted OR = 2.71, 95% CI = 1.01-7.24). Stratification analysis revealed that the increased risk was mainly confined to lung squamous cell carcinoma, with the ORs being 20.50 (95% CI = 2.25-179.05) for the 312Asn/Asn genotype and 4.24 (95% CI = 1.34-13.38) for the 751Gln/Gln genotype, respectively. Haplotype analysis with the 2 polymorphisms suggested these polymorphisms might be in linkage disequilibrium with a different causative locus or act together with other functional variants in or close to the XPD locus. PMID- 12740917 TI - Differential prognostic impact of the cyclins E and B in premenopausal and postmenopausal women with lymph node-negative breast cancer. AB - Searching for new prognostic factors, we investigated the influence of cyclin expression on breast cancer prognosis. A total of 273 archival tumor specimens from patients with pT1/pT2 N0 breast cancers treated by surgery and local irradiation were immunostained for cyclins E, A and B. Outcome was evaluated as metastasis-free (MFS) and disease-specific survival (DSS) over a median observation period of 99 months. In postmenopausal women, DSS was significantly predicted by cyclin E, and in premenopausal patients by cyclin B. No statistical significance was found for cyclin A. When the prognostic impact of cyclins was compared to that of standard prognostic indicators in a multivariate analysis, both cyclin E and cyclin B were selected as independent predictors of survival in postmenopausal and premenopausal patients, respectively. After inclusion of Ki-67 in the model, cyclin E lost its significance, whereas cyclin B remained the only independent prognostic factor with a hazard ratio of 4.5 (p = 0.026) for tumor related death. Assessment of cyclin expression may, therefore, refine current prognostic models if considered in relation to menopausal status. The prognostic relevance of cyclins is likely attributable to an influence on proliferation, cell survival and genetic instability. Awareness of the molecular mechanisms leading to deregulated cyclin expression may guide decisions for risk-adapted therapy regimens. PMID- 12740918 TI - Independent and combined effects of tobacco smoking, chewing and alcohol drinking on the risk of oral, pharyngeal and esophageal cancers in Indian men. AB - Oral, pharyngeal and esophageal cancers are 3 of the 5 most common cancer sites in Indian men. To assess the effect of different patterns of smoking, chewing and alcohol drinking in the development of the above 3 neoplasms and to determine the interaction among these habits, we conducted a case-control study in Chennai and Trivandrum, South India. The cases included 1,563 oral, 636 pharyngeal and 566 esophageal male cancer patients who were compared with 1,711 male disease controls from the 2 centers as well as 1,927 male healthy hospital visitors from Chennai. We observed a significant dose-response relationship for duration and amount of consumption of the 3 habits with the development of the 3 neoplasms. Tobacco chewing emerged as the strongest risk factor for oral cancer, with the highest odds ratio (OR) for chewing products containing tobacco of 5.05 [95% confidence internal (CI) 4.26-5.97]. The strongest risk factor for pharyngeal and esophageal cancers was tobacco smoking, with ORs of 4.00 (95% CI 3.07-5.22) and 2.83 (95% CI 2.18-3.66) in current smokers, respectively. An independent increase in risk was observed for each habit in the absence of the other 2. For example, the OR of oral cancers for alcohol drinking in never smokers and never chewers was 2.56 (95% CI 1.42-4.64) and that of esophageal cancers was 3.41 (95% CI 1.46 7.99). Furthermore, significant decreases in risks for all 3 cancer sites were observed in subjects who quit smoking even among those who had quit smoking 2-4 years before the interview. PMID- 12740919 TI - Meta-analysis of social inequality and the risk of cervical cancer. AB - Previous studies of the relationship between socio-economic status and cervical cancer have been mainly based on record linkage of routine data, such as cancer registry incidence rates and regional measures of social class based on census data. These routine data are liable to substantial misclassification with respect to socio-economic status. Previous reports are also primarily from developed countries, whereas the major burden of cervical cancer is in developing countries. We have therefore pooled the data from previously reported case control studies of cervical cancer or dysplasia, which contain individual-level information on socio-economic characteristics to investigate the relationship between cervical cancer, social class, stage of disease, geographical region, age and histological type. Based on 57 studies, we found an increased risk of approximately 100% between high and low social class categories for the development of invasive cervical cancer, and an increased risk of approximately 60% for dysplasia, including carcinoma in situ. Although the difference was observed in all countries, it was stronger in low/middle income countries and in North America than in Europe. No clear differences were observed between squamous cell carcinoma and adenocarcinoma, or between younger and older women. These results indicate that both cervical infection with human papillomavirus, which is linked to both female and male sexual behaviour, and access to adequate cervical cancer screening programmes are likely to be important in explaining the large cervical cancer incidence rates observed in different socio-economic groups, and that the importance of these factors may vary between different geographical regions. PMID- 12740920 TI - Socioeconomic factors in cancer in Sweden. AB - It is well known that certain cancers have shown clustering in socioeconomic groups, but limited data are available on recent results and time trends in such clustering. We determined standardized incidence ratios (SIR) for cancer, adjusted for age, period, region, parity and age at first childbirth among men and women in 6 socioeconomic groups based on the Swedish Family-Cancer Database. Persons had to be identified with the same socioeconomic status in the census of years 1960 and 1970, or of years 1960, 1970 and 1980; the comparison group was all people according to the same censuses. Cancers were followed from years 1970 to 1998 or from 1980 to 1998. Both increased and decreased SIRs were found, and a consistent pattern emerged, although the overall SIRs for cancer did not differ much, the lowest being for farmers (0.85) and the highest for professional men (1.07) and women (1.11). At individual sites, manual workers were at risk of tobacco-, alcohol- and occupation- and human papilloma virus-related cancers and at a decreased risk at most other cancers. Manual workers and farmers showed an excess of stomach cancer; professionals had an excess of melanoma and squamous cell skin cancer. Male and female SIRs correlated highly for manual and blue collar workers and for professionals. The overall population-attributable fraction for selected sites was 16.7% for men and 10.9% for women and it was highest, over 50%, for lung cancer in both genders. PMID- 12740921 TI - Familial invasive and borderline ovarian tumors by proband status, age and histology. AB - Age-specific familial risks in ovarian cancer have not been assessed by histologic types of medically verified cancers. We used the nationwide Swedish Family-Cancer Database on 10.2 million individuals and 19,175 invasive and 3,436 borderline ovarian cancers to calculate, by affected family members, standardized incidence ratios (SIRs) and 95% confidence intervals (CIs) for familial ovarian cancer in 0-66 year old daughters. SIRs for all invasive ovarian cancer were 2.68 (95% CI 2.22-3.21) by ovarian cancer in mother, 2.94 (1.40-5.94) by an affected sister and 24.03 (6.12-74.46) by both an affected mother and sister. The population-attributable fraction from mothers was 2.52%. Seropapillary cystadenocarcinoma showed the highest familial risk, but the effect of histopathol subtype could not be fully assessed because of lack of data in probands. Age-specific data showed some early-onset components and an unusual maximal incidence in the 40s. A comparison to an earlier study on BRCA1/2 mutation analysis and relative risks of ovarian and breast cancer suggests that these mutations could account for 26% of the familial aggregation of ovarian cancer. Histopathology and age of onset appear to be important attributes of familial ovarian cancer, suggesting that further gene identification efforts should target a specific histopathology in early-onset patients. PMID- 12740922 TI - EBV specific antibody-based and DNA-based assays in serologic diagnosis of nasopharyngeal carcinoma. AB - We assessed 5 EBV specific assays for their capacity to effect serologic diagnosis of suspected NPC. The assays were the immunofluorescent assays, VCA IgA and EA IgA, the enzyme-linked immunosorbent assays specific for EBNA 1 IgA or zta IgG and an EBV DNA assay. Serum samples were taken from 218 symptomatic NPC patients presenting consecutively at a public hospital in Hong Kong, 51 of whom were subsequently diagnosed as having NPC; 4 had EBV-associated lung cancer with similar serology as NPC. The remaining patients included 23 who had other cancers and 140 who had other diseases. Objectives of serodiagnosis under such clinical settings, therefore, are to both exclude and predict a diagnosis of NPC. None of the assays individually can meet both requirements adequately, however. The difficulty was best overcome by combining EBNA 1 IgA and zta IgG. It was shown that 68.3% of the patients gave a confirmed test results, negative or positive, by both tests. A confirmed negative result was associated with a negative predictive value of 99.1%, providing a clear indication to exclude a diagnosis of NPC; a confirmed positive result was associated with a positive predictive value of 86.8%, providing a clear indication to proceed with diagnostic work-up of NPC. The remaining patients gave equivocal test results, being positive for one or the other test, which were associated with a positive predictive value of 43.3% and 24.2%, respectively. PMID- 12740923 TI - Antisense treatment against Ki-67 mRNA inhibits proliferation and tumor growth in vitro and in vivo. AB - The Ki-67 protein is tightly regulated and depends on the proliferative status of a cell. It is present in the nuclei of proliferating cells but absent in resting cells. Since transformation of malignant cells is frequently associated with high cell proliferation and since proliferation is tightly associated with the Ki-67 protein labeling index, this antigen may represent a potential target for cancer therapy. In the present study we determined the ability of a phosphorothioate antisense oligodeoxyribonucleotide (ODN) targeted against Ki-67 mRNA to inhibit tumor cell proliferation specifically in cell culture, in multicellular 3 dimensional spheroids (MCS) and in subcutaneous murine tumor models. Antisense treatment of 1 myeloid and different epithelial tumor cell lines in suspension and monolayer culture, respectively, resulted in specific reduction of Ki-67 mRNA and protein, inhibition of proliferation and increased apoptotic cell death. Multicellular human bladder carcinoma spheroids lost their 3-dimensional structure and underwent cell death after incubation with antisense oligonucleotides. The growth of subcutaneous syngeneic prostatic (p = 0.05) and transitional cell tumors (p = 0.001) in immunocompetent mice was significantly inhibited in antisense-treated animals. From these findings we conclude that antisense inhibition of Ki-67 protein expression may be a rational approach in anticancer therapy. PMID- 12740924 TI - Natural killer (NK) cell function is a strong prognostic factor in colorectal carcinoma patients treated with the monoclonal antibody 17-1A. AB - Tumor cells might be susceptible to different effector functions of the immune system. This cytotoxic capacity has been utilized to analyze the prognostic significance of peripheral blood mononuclear cells (PBMC) in patients with metastatic colorectal carcinoma (CRC) treated with the monoclonal antibody (MAb)17-1A. Such analysis might form the basis for future patient selection and may lead to improvements in therapeutic strategies. Between 1986 and 1998, 73 patients were treated with regimens containing MAb17-1A. Prior to therapy, the lytic capability of PBMC was assayed against: K562 (4 hr assay), the CRC cell line SW948 (4 hr and 18 hr assays) and antibody-dependent cellular cytotoxicity (ADCC, 18 hr assay). Since the study was performed over 13 years, the assays were checked for time-related bias. Reproducibility over time was satisfactory. Patients exhibited a significantly higher cytotoxic capability in all 4 assays compared to healthy control donors. No correlation to clinical outcome was noted for 18 hr ADCC and 18 hr spontaneous cytotoxicity. Pretreatment natural killer (NK) cell cytotoxicity (K562) was significantly related to overall survival (OS), progression-free survival (PFS), and response rate. OS for patients with high and low NK cell cytotoxicity was 71 vs. 30 weeks, respectively (p = 0.007). NK cell cytotoxicity (K562) was an independent prognostic factor for OS (p = 0.016). Pretreatment NK cell activity is a strong prognostic factor for patients with metastatic CRC receiving MAb17-1A therapy and is a predictor for OS, PFS and response. These results should be considered when designing antibody-based therapeutic protocols. PMID- 12740925 TI - Clinical significance of detecting elevated serum DcR3/TR6/M68 in malignant tumor patients. AB - TR6/DcR3/M68 is a soluble receptor that belongs to the TNF receptor family. It is expressed in malignant cells of several tumor types and has been postulated to help tumor cells to gain survival advantage by inhibiting apoptosis and by interfering with immune surveillance. In our study, we assessed for the first time serum TR6 in tumor patients to explore its diagnostic and prognostic value. We examined serum TR6 levels with ELISA in 146 tumor patients, 19 patients with acute infection, 5 patients with liver cirrhosis and 29 healthy individuals. TR6 expression in tumor mass was studied with immunohistochemistry. TR6 gene copy number in tumor tissues was evaluated by real time PCR. Ninety-seven point nine percent (47 of 48 cases) of healthy individuals and patients with acute infection were serum TR6-negative. In contrast, 56.2% (82 of 146 cases) of the tumor patients were serum TR6-positive. Almost all serum TR6-positive individuals (98.8%, 82 out of 83 cases) had malignancy, excluding the cases of liver cirrhosis. In gastric carcinomas, serum TR6 levels were closely correlated with tumor differentiation status and TNM classification. Tumor mass was the source of serum TR6 because its levels decreased drastically after curative tumor resection. TR6 gene amplification occurred in about half of liver carcinomas, but not in gastric or pancreatic carcinomas, indicating plural mechanisms of TR6 upregulation. Our study demonstrated that serum TR6 should be considered as a novel parameter for the diagnosis, treatment and prognosis of malignancies. PMID- 12740927 TI - Optimization of catechol production by membrane-immobilized polyphenol oxidase: a modeling approach. AB - Although previous research has focused on phenol removal efficiencies using polyphenol oxidase in nonimmobilized and immobilized forms, there has been little consideration of the use of polyphenol oxidase in a biotransformation system for the production of catechols. In this study, polyphenol oxidase was successfully immobilized on various synthetic membranes and used to convert phenolic substrates to catechol products. A neural network model was developed and used to model the rates of substrate utilization and catechol production for both nonimmobilized and immobilized polyphenol oxidase. The results indicate that the biotransformation of the phenols to their corresponding catechols was strongly influenced by the immobilization support, resulting in differing yields of catechols. Hydrophilic membranes were found to be the most suitable immobilization supports for catechol production. The successful biocatalytic production of 3-methylcatechol, 4-methylcatechol, catechol, and 4-chlorocatechol is demonstrated. PMID- 12740928 TI - A novel biphasic extractive membrane bioreactor for minimization of membrane attached biofilms. AB - Extractive membrane bioreactor (EMB) systems offer a means of biologically treating wastewaters, but, like other membrane processes, are constrained by their tendency to be fouled by membrane-attached biofilms (MABs). This study describes a new approach to eradicate MAB formation and accumulation in EMB systems. To this end, an innovative EMB configuration, the biphasic extractive membrane bioreactor (BEMB), has been developed. In BEMB systems, the two main constituents of the EMB process, membrane and bacteria, are kept separated and interact via a suitable recirculating solvent. Nineteen candidate solvents were tested to assess their suitability for BEMB application. Based on the results of the solvent selection, guidelines are provided to screen solvents for BEMB application. BEMB and EMB runs were carried out to demonstrate the effectiveness of BEMB technology in avoiding MAB accumulation and to compare BEMB and EMB performance. A synthetic wastewater containing monochlorobenzene (MCB) was used as a model system. Abiotic BEMB and EMB runs were carried out and used as comparative references for estimating the effect of MAB accumulation on system performance. MAB thickness in the BEMB systems was controlled at 18 microm during 1 month of operation, whereas, in the EMB systems, MAB thickness reached 1250 microm. Analysis of mass transport in EMB and BEMB systems revealed that the high affinity of the permeating molecules for the solvent may contribute to a reduction in shell-side mass transfer resistance. This reduction of shell-side mass transfer resistance and the absence of MAB accumulation led to overall mass transfer coefficients of about sevenfold greater (4.5 x 10(-5) m s(-1)) in the BEMB system than in the EMB system (0.6 x 10(-5) m s(-1)). PMID- 12740929 TI - Multienzyme catalysis in microfluidic biochips. AB - The attachment of enzymes to glass microfluidic channels has been achieved using a highly reactive poly(maleic anhydride-alt-alpha-olefin) (PMA)-based coating that is supplied to the microchannel in a toluene solution. The PMA reacts with 3 aminopropyltriethoxysilane groups linked to the glass surface to form a matrix that enables additional maleic anhydride groups to react with free amino groups on enzymes to give a mixed covalent-noncovalent immobilization support. Using a simple T-channel microfluidic design, with reaction channel dimensions of 200 microm wide (at the center), 15 microm deep, and 30 mm long giving a reaction volume of 90 nL, soybean peroxidase (SBP) was attached at an amount up to 0.6 microg/channel. SBP-catalyzed oxidation of p-cresol was performed in aqueous buffer (with 20% [v/v], dimethylformamide) containing H(2)O(2), with microfluidic transport enabled by electroosmotic flow (EOF). Michaelis-Menten kinetics were obtained with K(m) and V(max) values of 0.98 mM and 0.21 micromol H(2)O(2) converted/mg SBP per minute, respectively. These values are nearly identical to nonimmobilized SBP kinetics in aqueous-DMF solutions in 20-microL volumes in 384 well plates and 5-mL reaction volumes in 20-mL scintillation vials. These results indicate that SBP displays intrinsically native activity even in the immobilized form at the microscale, and further attests to the mild immobilization conditions afforded by PMA. Bienzymic and trienzymic reactions were also performed in the microfluidic biochip. Specifically, a combined Candida antarctica lipase B-SBP bienzymic system was used to convert tolyl acetate into poly(p-cresol), and an invertase-glucose oxidase SBP trienzymic system was used to take sucrose and generate H(2)O(2) for SBP-catalyzed synthesis of poly(p-cresol). PMID- 12740931 TI - Construction of biofilms with defined internal architecture using dielectrophoresis and flocculation. AB - A novel approach was developed for the construction of biofilms with defined internal architecture using AC electrokinetics and flocculation. Artificial structured microbial consortia (ASMC) consisting of localized layered microcolonies of different cell types were formed by sequentially attracting different cell types to high field regions near microelectrodes using dielectrophoresis. Stabilization of the microbial consortia on the electrode surface was achieved by crosslinking the cells using the flocculant polyethyleneimine (PEI). Consortia of Escherichia coli, Micrococcus luteus, and Saccharomyces cerevisiae were made as model systems. Also, more natural consortia were made of the bacteria Pseudomonas putida, Clavibacter michiganense, and Methylobacterium mesophilum, which are found together in consortia during biodegradation of metal-cutting waste fluids. PMID- 12740930 TI - Macrokinetic and quantitative microbial investigation on a bench-scale biofilter treating styrene-polluted gaseous streams. AB - We performed a macrokinetic and quantitative microbial investigation of a continuously operating bench-scale biofilter treating styrene-polluted gases. The device was filled with a mixture of peat and glass beads as packing medium and inoculated with the styrene-oxidizing strain, Rhodococcus rhodochrous AL NCIMB 13259. The experimental data of styrene and microbial concentrations, obtained at different biofilter heights, were used to evaluate the pollutant concentration profiles as well as the influence of styrene loading on biomass distribution along the packing medium. Styrene and biomass concentration profiles permitted detection of a linear relationship between the amount of biomass grown in a given section of the biofilter and that of pollutant removed, regardless of the operating conditions tested. Biomass development in the bed appeared to: depend linearly on pollutant concentration at an inlet styrene concentration of <0.10 g m(-3) in the gaseous stream; achieve a maximum value (7. 10(7) colony forming units per gram of packing material) within a wide styrene concentration range (0.10 to 1.0 g m(-3)); and fall sharply beyond this inhibition threshold. The process followed zeroth-order macrokinetics with respect to styrene concentration, which is consistent with zeroth-order microkinetics with either fully active or not fully active biofilm. The maximal volumetric styrene removal rate was found to be 63 g m(packing material) (-3) h(-1) for an influent pollutant concentration of 0.80 g m(-3) and a superficial gas velocity of 245 m h(-1). PMID- 12740932 TI - Large-scale propagation of a replication-defective adenovirus vector in stirred tank bioreactor PER.C6 cell culture under sparging conditions. AB - Large-scale propagation of replication-defective adenovirus vectors has not been well studied to date. One of the challenges for efficient propagation at large scale is to overcome the sensitivity of virus infected cells to gas sparging required for oxygenation and CO(2) removal. In our initial experiments, it was observed that productivity of an adenovirus vector was significantly reduced under sparging conditions as compared to nonsparged, i.e., surface-aerated controls in serum-free cultures. Investigations led to the identification of a buffer containing surfactant (Polysorbate-80, PS-80) that was included in the virus seed stock formulation and introduced through virus infection into the culture at a very low concentration as the cause of the reduced virus productivity. This finding was not obvious and trivial, as neither uninfected sparged nor infected nonsparged PER.C6 trade mark cells in serum-free cultures were affected by the buffer at such a low PS-80 concentration of 0.00025% (v/v), which is a common component of serum-free cell culture media. These results strongly suggest that virus-infected cells behave very differently from uninfected cells under sparging conditions. To mitigate the deleterious effects of sparging, the virus seed stock was prepared in the absence of the buffer containing PS-80. At the same time, the concentration of Pluronic-F68 (PF-68) in the serum-free medium was increased to 1 g/L, at which cell growth and metabolism were unaffected, even though this measure alone did not result in virus productivity improvement. Only by implementing the two measures together was virus productivity loss completely eliminated under sparging conditions. After demonstration of the process robustness in 2-L bioreactors, this adenovirus propagation process was successfully scaled up to 250 L in a 300-L bioreactor under the worst-case sparging conditions projected for 10,000-L scale. PMID- 12740933 TI - Metabolic load of recombinant protein production: inhibition of cellular capacities for glucose uptake and respiration after induction of a heterologous gene in Escherichia coli. AB - The strong expression of recombinant proteins in bacteria affects the primary carbon and energy metabolism resulting in growth inhibition and acetate formation. By applying glucose pulses to fed-batch fermentations performed for production of a heterologous (alpha-glucosidase in Escherichia coli, we show that the induction of the recombinant gene strongly inhibits the maximum specific uptake capacities for glucose and the respiration capacity. The accumulation of glucose in the fermentation medium promotes the growth of plasmid-free cells. These inhibition effects are well described by including the kinetics of product formation into a recently published dynamic model (Lin et al. [2001] Biotechnol Bioeng 73:349-357). The new model also includes the population characteristics and gives a good fit to the measured data describing growth, production, substrate consumption, by-product formation, and respiration. PMID- 12740934 TI - Fluid shear contributions to bacteria cell detachment initiated by a monoclonal antibody. AB - Receptor-mediated adhesion of bacteria to biological surfaces is a significant step leading to infection. Due to an increase in bacterial antibiotic resistance, novel methods to block and disrupt these specific interactions have gained considerable interest as possible therapeutic strategies. Recently, several monoclonal antibodies specific for the Staphylococcus aureus collagen receptor demonstrated specialized ability to displace attached cells from collagen in static assays. In this study, we experimentally examine the monoclonal antibody detachment functionality under physiological shear conditions to evaluate the role of this parameter in the detachment process. The detachment of staphylococci from collagen was quantified in real-time using a parallel plate flow chamber, phase contrast video-microscopy and digital image processing. The results demonstrate a unimodal dependence of detachment on fluid wall shear rate. The observed decrease in effective detachment rate with increasing force at the highest shear levels evaluated is counterintuitive and has not been previously demonstrated. Several possible mechanisms of this result are discussed. PMID- 12740935 TI - Metabolic flux and metabolic network analysis of Penicillium chrysogenum using 2D [13C, 1H] COSY NMR measurements and cumulative bondomer simulation. AB - At present two alternative methods are available for analyzing the fluxes in a metabolic network: (1) combining measurements of net conversion rates with a set of metabolite balances including the cofactor balances, or (2) leaving out the cofactor balances and fitting the resulting free fluxes to measured (13)C labeling data. In this study these two approaches are applied to the fluxes in the glycolysis and pentose phosphate pathway of Penicillium chrysogenum growing on either ammonia or nitrate as the nitrogen source, which is expected to give different pentose phosphate pathway fluxes. The presented flux analyses are based on extensive sets of 2D [(13)C, (1)H] COSY data. A new concept is applied for simulation of this type of (13)C-labeling data: cumulative bondomer modeling. The outcomes of the (13)C-labeling based flux analysis substantially differ from those of the pure metabolite balancing approach. The fluxes that are determined using (13)C-labeling data are shown to be highly dependent on the chosen metabolic network. Extending the traditional nonoxidative pentose phosphate pathway with additional transketolase and transaldolase reactions, extending the glycolysis with a fructose 6-phosphate aldolase/dihydroxyacetone kinase reaction sequence or adding a phosphoenolpyruvate carboxykinase reaction to the model considerably improves the fit of the measured and the simulated NMR data. The results obtained using the extended version of the nonoxidative pentose phosphate pathway model show that the transketolase and transaldolase reactions need not be assumed reversible to get a good fit of the (13)C-labeling data. Strict statistical testing of the outcomes of (13)C-labeling based flux analysis using realistic measurement errors is demonstrated to be of prime importance for verifying the assumed metabolic model. PMID- 12740936 TI - The use of dinoflagellate bioluminescence to characterize cell stimulation in bioreactors. AB - Bioluminescent dinoflagellates are flow-sensitive marine organisms that produce light emission almost instantaneously upon stimulation by fluid shear in a shear stress dose-dependent manner. In the present study we tested the hypothesis that monitoring bioluminescence by suspended dinoflagellates can be used as a tool to characterize cellular response to hydrodynamic forces in agitated bioreactors. Specific studies were performed to determine: (1) impeller configurations with minimum cell activation, (2) correlations of cellular response and an integrated shear factor, and (3) the effect of rapid acceleration in agitation. Results indicated that (1) at a volumetric mass transfer coefficient of 3 x 10(-4) s(-1), marine impeller configurations were less stimulatory than Rushton configurations, (2) bioluminescence response and a modified volumetric integrated shear factor had an excellent correlation, and (3) rapid acceleration in agitation was highly stimulatory, suggesting a profound effect of temporal gradients in shear in increasing cell stimulation. By using bioluminescence stimulation as an indicator of agitation-induced cell stimulation and/or damage in microcarrier cultures, the present study allows for the verification of hypotheses and development of novel mechanisms of cell damage in bioreactors. PMID- 12740937 TI - Evaluation of spectrofluorometry as a tool for estimation in fed-batch fermentations. AB - Native culture fluorescence was investigated as an additional source of information for predicting biomass and glucose concentrations in a fed-batch fermentation of Alcaligenes eutrophus. Partial least squares (PLS) regression and a feed forward neural network (FFNN) coupled with principle component analysis (PCA) were each used to model the kinetics of the fermentation. Data from three fermentations was combined to form a training set for model calibration and data from a fourth fermentation was used as the testing set. The fluorescent soft sensors were compared with a previously developed feed forward neural network soft-sensor model which used oxygen uptake rate (OUR), carbon dioxide evolution rate (CER), aeration rate, feed rate, and fermentor volume to estimate biomass and glucose concentrations. The best model performance for predicting both biomass and glucose concentrations was achieved using the native fluorescence based models. Real data predictions of the biomass concentration in the testing set were obtained using both the PLS and FFNN PCA modeling utilizing fluorescence measurements plus the rate of change of the fluorescence measurements. Accurate predictions of the glucose concentration in the testing set were obtained using the FFNN PCA modeling technique utilizing the rate of change of the fluorescence measurements. Substrate exhaustion was indicated qualitatively by a first-order PLS model utilizing the rate of change of fluorescence measurements. These results indicate that native culture fluorescence shows promise for providing additional valuable information to enhance predictive modeling which cannot be extracted from other easily acquired measurements. PMID- 12740939 TI - Release kinetics of transforming growth factor-beta1 from fibrin clots. AB - In any therapeutic model involving a tissue-engineering approach to the repair of partial-thickness articular cartilage defects, a chondrogenic differentiation factor is required to ensure tissue-specific healing. Transforming growth factor beta1 (TGF-beta1) is known to act in this capacity, but at such high concentrations as to render its direct injection into the joint cavity inadvisable. This situation calls for a delivery system that can be applied directly to the defect site and that will release the drug gradually over a period of some weeks. Liposome encapsulation represents one such system, and has been recently implemented with some success in an animal model for cartilage repair. However, the kinetics of TGF-beta1 release have not been determined, it was the purpose of the present study to characterize these. The liberation of [(125)I]-labeled TGF-beta1 from fibrin matrices containing this agent in either a free or liposome-encapsulated form was monitored by liquid scintillation counting for 25 days in vitro. During the initial 5 days, fibrin clots containing liposome encapsulated TGF-beta1 released this cytokine at a slower rate (2% to 4% per day) than did those containing the free molecules (10% to 20% per day); thereafter, the release rates were similar. At the end of the incubation period, only 40% of the liposome-encapsulated TGF-beta1 had been released from the fibrin clots, as compared with 68% from those containing the free molecules. Liposome encapsulation thus represents a suitable means of establishing a slow-delivery system in tissue-engineering approaches to articular cartilage repair. PMID- 12740938 TI - Hydrodynamic stress and lethal events in sparged microalgae cultures. AB - The effect of high superficial gas velocities in continuous and batch cultures of the strains Dunaliella tertiolecta, Chlamydomonas reinhardtii wild-type and cell wall-lacking mutant was studied in bubble columns. No cell damage was found for D. tertiolecta and C. reinhardtii (wild-type) up to superficial gas velocities of 0.076 and 0.085 m s(-1), respectively, suggesting that high superficial gas velocities alone cannot be responsible for cell death and, consequently, bubble bursting cannot be the sole cause for cell injury. A death rate of 0.46 +/- 0.08 h(-1) was found for C. reinhardtii (cell wall-lacking mutant) at a superficial gas velocity of 0.076 m s(-1), and increased to 1.01 +/- 0.29 h(-1) on increasing superficial gas velocity to 0.085 m s(-1). Shear sensitivity is thus strain dependent and to some extent the cell wall plays a role in the protection against hydrodynamic shear. When studying the effect of bubble formation at the sparger in batch cultures of D. tertiolecta by varying the number of nozzles, a death rate of 0.047 +/- 0.016 h(-1) was obtained at high gas entrance velocities. D. tertiolecta was cultivated in a pilot-plant reactor under different superficial gas velocities of up to 0.026 m s(-1), with relatively low gas entrance velocities and no cell damage was observed. There is some indication that the main parameter causing cell death and damage was the gas entrance velocity at the sparger. PMID- 12740940 TI - Morphologies and projections of defined classes of neurons in the submucosa of the guinea-pig small intestine. AB - Four types of neurons have previously been identified by neurochemical markers in the submucosal ganglia of the guinea-pig small intestine, and functional roles have been ascribed to each type. However, morphological differences among the classes have not been determined, and there is only partial information about their projections within the submucosa. In the present work, we used intracellular microelectrodes to fill neurons of each type with biocytin, which was then converted to a permanent dye, so that the shapes of the neurons could be determined and their projections within the submucosa could be followed. Cell bodies of noncholinergic secretomotor/ vasodilator neurons had Dogiel type I morphology. These neurons, which are vasoactive intestinal peptide immunoreactive, had single axons that ran through many ganglia without providing terminals around other neurons. Cholinergic secretomotor neurons with neuropeptide Y immunoreactivity had Stach type IV morphology, and cholinergic secretomotor/vasodilator neurons had stellate cell bodies. The axons of these two types ran short distances in the plexus and did not innervate other submucosal neurons. Neurons of the fourth type, intrinsic primary afferent neurons, had cell bodies with Dogiel type II morphology and their processes supplied networks of varicose processes around other nerve cells. It is concluded that each functionally defined type of submucosal neuron has a characteristic morphology and that intrinsic primary afferent neurons synapse with secretomotor neurons to form monosynaptic secretomotor reflex circuits. PMID- 12740941 TI - Giant mitochondria in the retina cone inner segments of shrews of genus Sorex (Insectivora, Soricidae). AB - The retinas of three species of shrews (Sorex araneus, S. coronatus, and S. minutus) were analyzed. Two kinds of photoreceptors were identified according to (among other characteristics) the traits of the mitochondria of their inner segments. The rod inner segments contained several round or oval mitochondria distributed longitudinally inside the ellipsoid. The cone inner segment showed a few mitochondria, which we classified as megamitochondria (maximum length = 4.22 microm in S. araneus, 5.68 microm in S. coronatus, and 2.42 microm in S. minutus). An analysis of serial thin sections in S. coronatus showed that these large organelles occurred in the apical and central portions of the ellipsoid. In the peripheral and basal regions of the ellipsoid, megamitochondria were frequently accompanied by smaller mitochondria. The giant mitochondria were irregular in form and densely packed, and a reduced cytosol was observed between each mitochondria. In general, they exhibited an electron-dense matrix and a complex system of cristae, which varied in length and array. In mammalian retina, megamitochondria have only been described in the ellipsoid of the tree shrews Tupaia glis and T. belangeri, two diurnal Scandentia with a rich-cone retina. In general terms, Sorex megamitochondria are morphologically very similar to those reported for Tupaia, especially in their arrangement in the cone ellipsoid. However, they differ in the orientation of the cristae. We propose that the ellipsoid of Sorex may serve two functions: as a source of energy for receptor cells, and as a device for improving the cone outer segment optics. PMID- 12740942 TI - Cell and organ printing 1: protein and cell printers. AB - We have developed several devices for positioning organic molecules, molecular aggregates, cells, and single-cell organisms onto solid supports. These printers can create stable, functional protein arrays using an inexpensive technology. The cell printer allows us to create cell libraries as well as cellular assemblies that mimic their respective position in organs. The printers are derived from commercially available ink-jet printers that are modified to dispense protein or cell solutions instead of ink. We describe here the modifications to the print heads, and the printer hardware and software that enabled us to adapt the ink-jet printers for the manufacture of cell and protein arrays. The printers have the advantage of being fully automated and computer controlled, and allow for the high-throughput manufacture of protein and cell arrays. PMID- 12740943 TI - Cell and organ printing 2: fusion of cell aggregates in three-dimensional gels. AB - We recently developed a cell printer (Wilson and Boland, 2003) that enables us to place cells in positions that mimic their respective positions in organs. However, this technology was limited to the printing of two-dimensional (2D) tissue constructs. Here we describe the use of thermosensitive gels to generate sequential layers for cell printing. The ability to drop cells on previously printed successive layers provides a real opportunity for the realization of three-dimensional (3D) organ printing. Organ printing will allow us to print complex 3D organs with computer-controlled, exact placing of different cell types, by a process that can be completed in several minutes. To demonstrate the feasibility of this novel technology, we showed that cell aggregates can be placed in the sequential layers of 3D gels close enough for fusion to occur. We estimated the optimum minimal thickness of the gel that can be reproducibly generated by dropping the liquid at room temperature onto a heated substrate. Then we generated cell aggregates with the corresponding (to the minimal thickness of the gel) size to ensure a direct contact between printed cell aggregates during sequential printing cycles. Finally, we demonstrated that these closely-placed cell aggregates could fuse in two types of thermosensitive 3D gels. Taken together, these data strongly support the feasibility of the proposed novel organ-printing technology. PMID- 12740944 TI - General suitability of techniques for in situ detection of apoptosis in small intestinal epithelium. AB - The present study was designed to evaluate different techniques for the in situ detection of apoptosis in human and rat small intestinal epithelium. The techniques included light microscopy (LM) and transmission electron microscopy (TEM) observation of epoxy resin-embedded tissue, scanning electron microscopy (SEM), TUNEL assay, and antibodies directed against caspase cleavage products of caspase 3, cytokeratin 18 (CK 18), and apoptotic single-strand DNA (ssDNA). All techniques, if the labeling was positive, showed apoptotic cells exclusively at the villus tip. LM and TEM were the most reliable and revealed morphological signs typical of cells that have died via apoptosis. SEM indicated the extension of the process. The antibody recognizing cleaved caspase 3 could be considered an appropriate marker for apoptotic epithelial cells in human and rat small intestine. However, the majority of epithelial cells lining the proximal small intestinal villus contained only low levels of intact CK 18. Therefore, sufficient amounts of cleaved CK 18 for immunohistochemical detection were not generated during apoptosis, rendering the application of the antibody inappropriate. The antibody detecting formamide-denatured ssDNA in apoptotic cells was both suitable and reliable; however, the particular staining procedure used compromised the tissue preservation. In comparison to this, the TUNEL assay was less reliable. Although it was performed with a commercially available ready to-use kit, its application conditions had to be adjusted for each specimen on the basis of the findings produced by other techniques. PMID- 12740945 TI - Development of the human Mullerian duct in the sexually undifferentiated stage. AB - An embryological explanation for the development of the Mullerian duct still poses a major challenge. The development of this duct was investigated systematically in human embryos. Seven embryos (Carnegie stages 18-23) were serially sectioned in the frontal, sagittal, and transversal planes at a thickness of 10 microm and stained with hematoxylin and eosin (H&E) for histological analysis. In all observed embryos, the caudal end of the Mullerian duct was found to be intimately connected to the Wolffian duct. The opening of the Mullerian duct to the coelomic cavity was formed as the result of an invagination of the coelomic epithelium at Carnegie stage 18. The duct grew independently from the invagination during stages 19-23. The fused duct (uterovaginal canal) bifurcated at the caudal portion at Carnegie stages 22 and 23. This is the first description of the caudal portion of the fused Mullerian ducts separating again and returning to each of the Wolffian ducts in human embryos. PMID- 12740946 TI - Histomorphometric assessment of Haversian canal and osteocyte lacunae in different-sized osteons in human rib. AB - There is no detailed information available concerning the variations in bone, the Haversian canal, and osteocyte populations in different-sized osteons. In this study a total of 398 secondary osteons were measured in archived rib sections from nine white men (20-25 years old). The sections were stained with basic fuchsin. The parameters included the osteon area (On.Ar), Haversian canal area (HC.Ar) and perimeter (HC.Pm), bone area (B.Ar), and osteocyte lacunar number (Lc.N). From these primary measurements the following indices were deduced: 1) lacunar number per bone area (Lc.N/B.Ar) and per osteon (Lc.N/On); 2) the ratio between Haversian canal perimeter and bone area (HC.Pm/B.Ar); and 3) the fraction of Haversian canal area (HC.Ar/On.Ar) and its complement, the fraction of bone area (B.Ar/On.Ar). The results showed that the osteons varied greatly in size, but very little in the fraction of bone area. Regression analyses showed that HC.Ar, HC.Pm, and Lc.N/On were positively associated with On.Ar (P < 0.001 for all). A significant negative correlation was found between On.Ar and Lc.N/B.Ar (P < 0.05) and HC.Pm/B.Ar (P < 0.0001). HC.Ar and HC.Pm increased significantly with increasing Lc.N/On (both P < 0.0001) rather than Lc.N/B.Ar. Lc.N/B.Ar had a significant positive correlation with HC.Ar/On.Ar (P < 0.05) and HC.Pm/B.Ar (P < 0.01). We conclude that: 1) the size of the osteon is determined by the quantum of bone removed by osteoclasts, 2) the osteon is well designed for molecular exchange, and 3) a well designed osteon may be produced via the regulation of bone apposition by osteocytes during the process of osteon refilling. PMID- 12740947 TI - Functional anatomy and hemodynamic characteristics of vasa vasorum in the walls of porcine coronary arteries. AB - In this study vasa vasorum in the walls of porcine coronary arteries were examined, using three-dimensional (3D) micro-CT scanning techniques. These techniques leave the 3D structure of the vasa vasorum tree intact and thus provide a much more direct view of this structure than is possible from conventional histological sections. The study demonstrates-for the first time, we believe-both the different types and the fine architecture of these vasa vasorum. Furthermore, with the use of automated tree analysis software, it was possible to obtain quantitative geometrical data on the 3D structure of vasa vasorum trees that have not previously been available. The results indicate that despite the restrictive topology of the space in which they are present, the branching architecture of the vasa vasorum trees, which we surveyed, is surprisingly similar to that of vasculature in general. The volume of vessel wall tissue perfused or drained by a vasa vasorum tree was found to correlate well with the cross-sectional area of the root segment of the vasa vasorum tree, and the luminal surface area corresponding to this volume was found to be comparable with the surface area of an early atherosclerotic lesion. This is consistent with earlier findings that the ligation or removal of vasa vasorum leads to atherogenesis. PMID- 12740949 TI - Rat anterior pituitary cells in vitro can partly reconstruct in vivo topographic affinities. AB - Hormone-producing cells in the rat anterior pituitary gland are not randomly distributed; rather, there are specific topographic affinities among five cell types (Noda et al., Acta Histochem. Cytochem. 2001;34:313-319). In this study we reconstructed these affinities, at least partially, in primary monolayer culture. Pituitary cells collected from adult male rats were enzymatically dispersed and cultured for 72 hr at a density of 1 x 10(5) cells/cm(2). We double-immunostained cells using antibodies against hormones, and then used confocal laser microscopy to examine the ability of the cells to attach to each other. We also statistically analyzed the affinity of all combinations of the five types of hormone-producing cells. We observed clusters by electron microscopy to identify junctional complexes between the cells. Confocal laser microscopy indicated that the features and attachment patterns of hormone-producing cells in vivo were similar to those in vitro. Statistical analyses revealed that the rates at which the five types of hormone-producing cells attached to growth hormone (GH)-, prolactin (PRL), and luteinizing hormone (LH)-producing cells were unequal, which suggests there are specific topographic affinities. The specific rates of adrenocorticotropic hormone (ACTH)-producing cell attachment to GH cells, LH to PRL cells, and PRL to LH cells were high, whereas that of PRL attachment to PRL cells was low. In addition, the rates correlated with the data from our previous in vivo study. Ultrastructural observations revealed few junctional complexes between hormone-producing cells. These results indicate that anterior pituitary hormone-producing cells can attach to specific types of cells by means of specific and/or nonspecific adhesion factors, and can reconstruct the topographic nature of the pituitary gland. PMID- 12740948 TI - Morphology of aging lung in F344/N rat: alveolar size, connective tissue, and smooth muscle cell markers. AB - This study investigated the morphological changes of lungs in F344/N rats (9-36 months old). We initially examined general and quantitative morphological changes, and then we used immunohistochemistry to detect distributional changes in collagen subtypes (types I, III, and IV) and smooth muscle cell (SMC) markers (alpha-smooth muscle actin (ASMA), gamma-smooth muscle actin (GSMA), desmin, and vimentin) in the lungs. In 24-month-old rats, alveolar ducts and alveolar sacs were enlarged, and alveoli were wider and shallower than in younger animals. In old rats (>/=27 months), terminal and respiratory bronchioles and alveolar ducts were dilated and alveoli were more extended than in 24-month-old rats. No age related distributional changes were observed for collagen types I, III, and IV as revealed by immunohistochemistry, or elastin as revealed by resorsin fuchsin. SMCs in the extra- and intrapulmonary bronchi were immunoreactive for ASMA, GSMA, and desmin, but not for vimentin at all ages. In old rats (>/=27 months), SMCs were loosely arranged in comparison with younger animals, and stainability for GSMA and desmin was decreased. In the respiratory bronchioles and alveolar ducts, a few cells immunoreactive for ASMA and vimentin were observed in the smooth muscle aggregations of the alveolar orifice in rats younger than 12 months. In older rats (>20 months), cells immunoreactive for ASMA and vimentin were increased in septal tips. In conclusion, extension of distal airways and immunohistochemical changes of SMC markers in F344/N rat lungs were evident by approximately 24 months of age, but there was no apparent change in connective tissue morphology. PMID- 12740950 TI - Neurocan in the embryonic avian heart and vasculature. AB - The chondroitin sulfate proteoglycan (CSPG) neurocan was previously considered to be nervous-system specific. However, we have found neurocan in the embryonic heart and vasculature. In stage 11 quail embryos, neurocan was prominently expressed in the myocardium, dorsal mesocardium, heart-forming fields, splanchnic mesoderm, and vicinity of the extraembryonic vaculature, and at lower levels in the endocardium. A comparison of neurocan staining with QH1 staining of vascular endothelial cells demonstrates that neurocan is frequently expressed by cells adjacent to endothelial cells, rather than by endothelial cells themselves. In some cases, a dispersed subset of cells are neurocan-positive in a field of cells that otherwise appear uniform in morphology. Later in development, neurocan expression becomes relatively limited to the nervous system. However, even in 10 day embryos, neurocan is expressed in the chorio-allantoic membrane in the tissue that separates closely packed, small-diameter blood vessels. In summary, our results suggest that neurocan may function as a barrier that regulates vascular patterning during development. PMID- 12740951 TI - Renal corpuscle of the sturgeon kidney: an ultrastructural, chemical dissection, and lectin-binding study. AB - The sturgeon is an ancient species of fish that thrives in a wide range of ecological environments, from freshwater to seawater. Basic in this process of adaptation is the ability of the kidney to control fluid filtration and urine formation. However, the morphological basis of this process is mostly unknown. The aim of the present study was to use microdissection techniques (scanning electron microscopy (SEM), transmission electron microscopy (TEM), and lectin binding histochemistry) to examine the structure of the renal corpuscle of the sturgeon Acipenser nacarii in order to reveal morphologic features that could be related to function, phylogeny, and habitat. The renal corpuscles are aligned along the intrarrenal arteries. The urinary pole shows a siphon-like neck segment (NS) in 92% of the nephrons, whose structural characteristics are different from those of other fish. The podocytes have cuboidal cellular bodies, intercellular contacts, and poorly developed cell processes. The podocyte glycocalyx contains N acetylglucosamine and lacks sialic acid. The structural and lectin-binding patterns are similar to those found in the immature mammalian kidney. The glomerular basement membrane (GBM) is very thick and consists of three layers: a lamina rara externa, a lamina densa, and a thick subendothelial lamina. The latter contains tubular microfibrils, collagen fibers, and long mesangial cell processes. Frequently, the podocyte bodies attach directly to the GBM, and the area occupied by the filtration slits is very small. Furthermore, the GBM shows a glycosylation pattern different from that observed in most vertebrates. Contrary to what would be expected in sturgeons living in freshwater, the A. nacarii renal corpuscle morphology suggests a low glomerular filtration rate. PMID- 12740952 TI - Mitochondrial DNA from pre-Columbian Ciboneys from Cuba and the prehistoric colonization of the Caribbean. AB - To assess the genetic affinities of extinct Ciboneys (also called Guanajuatabeys) from Cuba, 47 pre-Columbian skeletal samples belonging to this group were analyzed using ancient DNA techniques. At the time of European contact, the center and east of Cuba were occupied by agriculturalist Taino groups, while the west was mainly inhabited by Ciboneys, hunter-gatherers who have traditionally been considered a relic population descending from the initial colonization of the Caribbean. The mtDNA hypervariable region I (HVR-I) and haplogroup-specific markers were amplified and sequenced in 15 specimens using overlapping fragments; amplification from second extractions from the same sample, independent replication in different laboratories, and cloning of some PCR products support the authenticity of the sequences. Three of the five major mtDNA Amerindian lineages (A, C, and D) are present in the sample analyzed, in frequencies of 0.07, 0.60, and 0.33, respectively. Different phylogenetic analyses seem to suggest that the Caribbean most likely was populated from South America, although the data are still inconclusive, and Central American influences cannot be discarded. Our hypothesis is that the colonization of the Caribbean mainly took place in successive migration movements that emanated from the same area in South America, around the Lower Orinoco Valley: the first wave consisted of hunter gatherer groups (ancestors of the Ciboneys), a subsequent wave of agriculturalists (ancestors of the Tainos), and a latter one of nomadic Carib warriors. However, further genetic studies are needed to confirm this scenario. PMID- 12740953 TI - Molecular analyses of the "Pharaos:" Feasibility of molecular studies in ancient Egyptian material. PMID- 12740954 TI - Brief communication: ancient DNA prospects from Sri Lankan highland dry caves support an emerging global pattern. AB - Recovery of ancient DNA has become an increasingly important tool in elucidating the origins of past populations and their relationships. Unfortunately, many human skeletal remains do not contain original DNA amplifiable by polymerase chain reaction (PCR). Amino-acid racemization has proven to be a useful predictor of ancient DNA results. We analyzed the relative levels of amino-acid preservation and racemization of human samples from two highland dry-cave sites in Sri Lanka, and found that amino-acid enantiomer ratios were inconsistent with successful authentic DNA recovery. A review of the literature reveals that these results are consistent with a global pattern of poor DNA preservation in the tropics. PMID- 12740955 TI - Pre-Columbian treponemal disease from 14th century AD Safed, Israel, and implications for the medieval eastern Mediterranean. AB - In 1912, 68 medieval crania were excavated from a cave at Safed in the eastern Mediterranean and brought to the United Kingdom. It is only recently that these skulls have been studied for evidence of disease. One adult individual demonstrates multiple lesions of the cranial vault, compatible with treponematosis. Radiocarbon dating suggests the year of death to be between 1290 1420 AD. This range equates to the mamluk period, just after the crusades. This is the oldest dated case of treponematosis in the Middle East, and the first to confirm its presence there before the epidemiologically important transatlantic voyage of Christopher Columbus. The finding has significant implications for our understanding of the introduction of the disease to the Middle East and of the medieval diagnosis of ulcerating skin conditions by medical practitioners in the Mediterranean world. PMID- 12740956 TI - Food procurement and tooth use in two sympatric lemur species. AB - This study of two lemur species (Lemur catta and Propithecus v. verreauxi) in Madagascar combines observations of food procurement and initial food placement in the mouth with comparisons to food toughness and external properties. Food toughness was hypothesized to play a decisive role in determining food placement during ingestion. It was found that tougher foods are generally ingested on the postcanines for all foods eaten. However, when leaves and fruits are analyzed separately, food size and shape, represented here by mass and food type, are more reliable predictors of initial food placement. Larger leaves and bulkier fruits and stalks are ingested posteriorly. Leaf toughness is not related to leaf size, though the toughness and size of the most commonly eaten fruits are correlated. Furthermore, ingestive food toughness, which is the maximum toughness, and "average" food toughness may make different mechanical demands on the masticatory apparatus that have consequences for jaw morphology. PMID- 12740957 TI - Longitudinal study of tuberculosis outcomes among immunologically naive Ache natives of Paraguay. AB - This study documents the course of a tuberculosis epidemic in an immunologically naive group of South American Indians within fewer than 20 years after first sustained contact with outsiders. Groups of Northern Ache (ah-CHAY) of eastern Paraguay were contacted and settled on reservations between 1971-1979. Not surprisingly, the Ache are very susceptible to tuberculosis, and the epidemiological characteristics of the disease are quite different from those of populations that have had tuberculosis for centuries. Within 6 years of the first detected case of tuberculosis among the Ache, the prevalence rate of active tuberculosis cases reached 18.2%, and of infected cases among adults, 64.6%, some of the highest rates ever reported for any human group. Remarkably, males and females are equally likely to have been diagnosed with active tuberculosis, Ache children between birth and 5 years of age are least vulnerable to tuberculosis, high nutritional and socioeconomic status do not decrease the risk of disease or infection, and children immunized with BCG are less responsive to tuberculin challenge than are other children. Moreover, similar to the Yanomamo, but unlike populations of European or African descent, a high percentage of Ache with active disease test negative on tuberculin challenge tests (purified protein derivative; PPD). These differences may be due to a high prevalence of diminished cell mediated immunity, and T-helper 2 dominance. We also hypothesize that these immunological characteristics, low genetic diversity, hostile intergroup interactions, and behavioral noncompliance to treatment protocols together contribute to the high rates of active disease observed. Existing tuberculosis control programs are poorly equipped to handle the impact of these causal complexities on the course of recent tuberculosis epidemics that have quickly spread throughout native communities of Latin America during the last decade. PMID- 12740958 TI - Hopi Indians, "cultural" selection, and albinism. AB - The incidence of albinism in Hopi Indians has been estimated as approximately 1 in 200 individuals. It has been suggested that "cultural" selection as the result of a mating advantage of males with albinism has been important in the maintenance of this high incidence. To examine this hypothesis quantitatively, a model that includes male-mating advantage, mutation, and viability selection is analyzed. In order to play an important role in the maintenance of the high incidence of albinism, the necessary mating advantage of males with albinism appears unrealistically high. However, if the extent of viability selection against individuals with albinism is not as large as previously assumed, the necessary amount of mating advantage is not as high. Other related aspects are also discussed here, such as the type of albinism in Hopi Indians and its impact, the conditions for a polymorphism with male-mating advantage and viability selection, and the time necessary to change the incidence of albinism either by the relaxation or institution of male-mating advantage. PMID- 12740959 TI - Genetic structure of Mediterranean populations revealed by Y-chromosome haplotype analysis. AB - The allelic variability at six Y-chromosome-specific polymorphisms (YAP, DYS19, DYS389-I, DYS390, DYS391, and DYS392) was used to generate male-specific haplotypes in 333 males representing 12 population samples from the region around the Mediterranean sea. Extreme interindividual variation was observed, as more than 160 distinct Y-chromosome variants could be defined as six-locus haplotypes. Concomitant with this high variability, low levels of population genetic structure were observed. In particular, a "core" of populations directly facing the north and the east of the Mediterranean basin, from the Middle East to the Italian Peninsula, was found to be genetically undifferentiated. This observation, supported by a reanalysis of Y-specific binary polymorphisms in the same populations, suggests that at least part of the male-specific gene pools of these populations has either a very recent common origin (that could be related with the Neolithic demic diffusion hypothesis), and/or that gene flow has played a significant role in shaping the patterns of genetic variability in this region. In agreement with both hypotheses, we found that the spatial distribution of DYS392 alleles revealed a marked differentiation between the East and the West of the Mediterranean area. Through the analysis of microsatellite variation, the time to the most recent common ancestor (TMRCA) of the YAP(+) sublineage 4 has been estimated. The estimations, based on two different data sets, turn out to be quite recent (7,000-11,000 YBP), suggesting that this lineage may have been first introduced into Southern Europe through Neolithic migrations from the Middle East. PMID- 12740960 TI - Lateral facial soft-tissue prediction model: analysis using Fourier shape descriptors and traditional cephalometric methods. AB - This study was designed to investigate the relationship between traditional skeletal cephalometric measurement and Fourier analysis of the lateral soft tissue profile. A random sample of 121 untreated subjects of European descent, with wide ranges of malocclusions and underlying facial patterns, was selected in the Orthodontic Unit at the University of Melbourne. Lateral cephalograms were available for all subjects. Both traditional lateral cephalometric analysis and Fourier soft-tissue profile analysis were carried out. Multivariate statistical analysis among 11 hard-tissue cephalometric measurements and the first 50 Fourier harmonics was then performed. This analysis formed the basis for a subsequently proposed soft-tissue prediction model. From this model, 50 predicted x- and y harmonics were generated for each subject in the total sample. Calculation of Pearson's correlation coefficients between the actual and predicted harmonics revealed strong relationships for many of the lower-order harmonics. To further test the model, the prediction-coefficients derived from all 121 subjects were then used to make predictions for the first 50 x- and y-harmonics for a subgroup of 10 independent test subjects. Once again, Pearson's correlations between the actual and predicted harmonics of the test model in the lower-order harmonics revealed strong associations. Superimposition of the actual and predicted soft tissue outlines, however, revealed that much actual detail in the region between the nose and the chin was still lost using the predicted Fourier harmonics. This suggests that soft-tissue prediction based on this Fourier test model, while already useful in Forensic facial reconstruction, may not yet be appropriate for useful diagnosis and planning in clinical disciplines. PMID- 12740961 TI - Kennewick and Luzia: lessons from the European Upper Paleolithic. PMID- 12740963 TI - Smallpox and smallpox vaccination. PMID- 12740964 TI - Smallpox and smallpox vaccination. PMID- 12740965 TI - Smallpox and smallpox vaccination. PMID- 12740966 TI - Smallpox and smallpox vaccination. PMID- 12740967 TI - Smallpox and smallpox vaccination. PMID- 12740968 TI - Serum retinol levels and fracture risk. PMID- 12740969 TI - Off-pump coronary bypass surgery. PMID- 12740970 TI - Off-pump coronary bypass surgery. PMID- 12740971 TI - The Kugelberg Lecture. Neurophysiological studies of collateral reinnervation in man. PMID- 12740972 TI - The Berger Lecture. Chronobiology of sleep/wake and of sleepiness/alertness states in normal and sleep disordered human subjects. PMID- 12740973 TI - The Adrian Lecture. Can EEG coherence help solve the binding problem? PMID- 12740974 TI - Clinical neurophysiology in the intensive care unit. PMID- 12740975 TI - Electromyography in the paediatric intensive care unit (ICU). PMID- 12740976 TI - Pediatric electromyography in the acute care setting. PMID- 12740977 TI - Acute polyneuropathies encountered in the intensive care unit. PMID- 12740978 TI - Neuromonitoring in the operating room and intensive care unit: an update. PMID- 12740979 TI - Continuous ICU EEG monitoring. PMID- 12740980 TI - Long-term EEG monitoring in neonatal and pediatric intensive care. PMID- 12740981 TI - Neurophysiology of anaesthesia. AB - Methods of clinical neurophysiology are important in studying basic problems of anaesthesia such as the problem of the depth of anaesthesia, Some of the problems of clinical neurophysiology in awake subjects, such as the generators of somatosensory evoked potentials or cognitive evoked potentials, may be resolved by recording during EEG suppression. Finally, the mechanisms by which some anaesthetics produce epileptic phenomena (others or even the same agents may suppress epileptic activity) can only be resolved by EEG and EP recordings in human subjects, and by simultaneous recordings of intracellular and extracellular potentials in animals. PMID- 12740982 TI - The clinical neurophysiology of diseases of neuromuscular transmission. PMID- 12740983 TI - Quantitation and standardization in EMG and neurography. PMID- 12740984 TI - Motor unit number estimation (MUNE) with nerve conduction studies. PMID- 12740985 TI - Electrophysiologic diagnostics of very early stages of ALS. PMID- 12740986 TI - Preferential involvement of the fast conducting corticospinal tracts in patients with ALS. PMID- 12740987 TI - Role of EMG in the evaluation of presumed myopathies in the era of DNA analysis. PMID- 12740988 TI - Pediatric clinical EMG challenge with bulbar dysfunction. PMID- 12740989 TI - Physiology and electromyography of swallowing and its disorders. PMID- 12740990 TI - Multi-channel surface EMG in clinical neurophysiology. PMID- 12740991 TI - TeleMedicine and Internet EMG. PMID- 12740992 TI - Recent developments in our understanding of motor control, corticomotoneuronal connections and the activation of motoneurons and motoneuronal pools. PMID- 12740993 TI - Synchronization of lower limb motor units in spastic patients. PMID- 12740994 TI - Reciprocal inhibition in hemiplegic patients--a longitudinal study. PMID- 12740995 TI - Long-lasting simultaneous activation of cortical and subcortical structures in movement preparation and execution. PMID- 12740996 TI - Spatiotemporal ERD/ERS patterns during voluntary movement and motor imagery. PMID- 12740997 TI - The Bereitschaftspotential as an electrophysiological tool for studying the cortical organization of human voluntary action. PMID- 12740998 TI - The thalamic contribution to the emergence of the readiness potential. PMID- 12740999 TI - Event-related desynchronization of the mu rhythm in extension and flexion finger movements. PMID- 12741000 TI - Anatomy and physiology of the autonomic regulation of intestine and the urogenital system. PMID- 12741001 TI - Clinical neurophysiology of pelvic floor disorders. PMID- 12741002 TI - Autonomic nervous system evaluation: diagnosis of rectal and lower gut dysfunction. AB - The main symptoms of autonomic dysfunction of the lower gut are diarrhoea, constipation and faecal incontinence, but these symptoms are not specific. The main diagnostic procedures in the evaluation of the lower gut are transit studies with radiopaque markers, hydrogen breath tests, tests for the differentiation between osmotic and secretory diarrhoea (fasting test and/or stool analysis for electrolytes and osmolality), anorectal manometry and EMG of the anal sphincters. PMID- 12741003 TI - Neurophysiologic pelvic floor testing. PMID- 12741004 TI - Erectile dysfunction--diagnostic approach and treatment options. PMID- 12741005 TI - Evaluation of bladder dysfunction. AB - A careful and detailed history and physical examination can give clues to a neurogenic bladder disturbance. Urine analysis should always be performed to exclude urinary tract infection. The most important basic investigation is ultrasound of the kidneys and of residual urine. If these investigations reveal suspicion of a dysfunction, elaborate urological investigation, including cystoscopy and urodynamic studies, is necessary for classification and appropriate therapy of a neurogeneous vesicourethral disorder. PMID- 12741006 TI - EEG in neonates. What does the neonatal EEG tell about prognosis? PMID- 12741008 TI - Seizures and pseudoseizures. PMID- 12741007 TI - Evolution of EEG and seizures in childhood. PMID- 12741009 TI - EEG changes and epilepsy in developmental dysphasia. PMID- 12741011 TI - Clinical use of magnetoencephalography. PMID- 12741010 TI - Source localization of interictal epileptiform spike potentials estimated with a dipole tracing method using surface and subdural EEG recordings. PMID- 12741012 TI - Topics in transcranial magnetic stimulation. PMID- 12741013 TI - Recent advances in clinical neurophysiology of vision. PMID- 12741014 TI - Field specific visual evoked potentials for assessment of peripheral field defect in a paediatric population. PMID- 12741015 TI - Electrophysiological correlates of dementia. PMID- 12741016 TI - Circadian rhythmicity: from basic science to clinical approach. PMID- 12741017 TI - Reducing the risk of sudden infant death. PMID- 12741018 TI - Gates to awakening in early development. PMID- 12741019 TI - Insomnia in children. PMID- 12741020 TI - Daytime behavioral correlates of awakenings and bedtime resistance in preschool children. PMID- 12741021 TI - Excessive sleepiness: clinical aspects. PMID- 12741022 TI - A contribution to pathophysiology of idiopathic hypersomnia. PMID- 12741023 TI - The treatment of narcolepsy. PMID- 12741024 TI - Rett's syndrome--spindle activity analysis in NREM sleep. PMID- 12741025 TI - Nervous system channelopathies. PMID- 12741026 TI - Guillain-Barre syndrome: an overview of current concepts. PMID- 12741027 TI - Mononeuropathies of infancy and childhood. PMID- 12741028 TI - Spondylotic cervical myelopathy: three aspects of the problem. PMID- 12741030 TI - Current aspects of pathogenesis and therapy of multiple sclerosis. PMID- 12741029 TI - Are motor evoked potentials (MEPs) helpful in the differential diagnosis of spondylotic cervical myelopathy (SCM)? PMID- 12741031 TI - Clinical neurophysiological approaches to neuromuscular fatigue. PMID- 12741032 TI - Neuronal ceroid lipofuscinoses in childhood. PMID- 12741033 TI - European trends in bioethics: medical research on persons unable to consent. PMID- 12741034 TI - Persistent vegetative state: clinical and ethical issues. PMID- 12741035 TI - The project of a European protocol on human genetics. PMID- 12741036 TI - Collaboration brings rapid improvement at SSM Health Care. PMID- 12741037 TI - Computerized patient record ties ambulatory centers together. PMID- 12741038 TI - Process improvement integrated with patient safety initiatives. PMID- 12741039 TI - Washington nursing homes enlist unlikely clinical allies in pain management efforts. PMID- 12741040 TI - Comparative data is a powerful motivator for improvement. PMID- 12741041 TI - Online doctor-patient communication tool saves time on phone. AB - Online tool enables patients to consult with physicians maore efficiently. The Medical Clinic of North Texas recently implemented an online communication tool that enables physicians and patients to communicate electronically, resulting in greater efficiency and improved patient care. PMID- 12741042 TI - Evidence-based medicine system helps physicians order the right tests. AB - Evidence-based medicine system helps physicians order the right tests. Physicians affiliated with Brigham and Women's Hospital can consult a medical expert that will help them decide which diagnostic imaging test has the greatest chance of determining what is wrong with their patient. The expert is not a person, however. PMID- 12741043 TI - Advocate's disease management program reduces readmissions for CHF and asthma. AB - Advocate's disease management program reduces readmissions for congestive heart failure (CHF) and asthma. Educating CHF and asthma patients while they are in the hospital, using standing orders that reflect national guidelines, and then providing nurse specialists to follow up with patients while they are in the outpatient setting is proving to be a winning combination for Advocate Health Care. PMID- 12741044 TI - Bush: will this be the era of "behealthioral havecare"? PMID- 12741045 TI - HIPAA privacy rules: what plans, providers must know. PMID- 12741046 TI - Relapse: road to failure or pathway to success? PMID- 12741047 TI - Life at liberty: the pursuit of quality. PMID- 12741048 TI - Offering solutions, for today's workplace. PMID- 12741049 TI - Moving the focus beyond healthcare companies demanding more from EAP work/life vendors. PMID- 12741050 TI - Private insurance: what has parity brought. PMID- 12741051 TI - "Outstanding facilities who are successful and manage to retain their directors of nursing -- some for years" . PMID- 12741052 TI - Falls.... A prelude to litigation. AB - For years long-term care has found itself vulnerable to law suits. This is still true today. On any given day, nursing homes and the staff employed by them are only one unhappy client away from receiving a notice of intent. In today's litigious world, nursing homes must have in place, effective policies and procedures to minimize their risk for a lawsuit. Part of managing the risk, is having a multi-disciplinary team in place that takes a proactive approach to patient safety. The particularly high risk patients for incident are ambulatory patients with cognitive impairment. The multi disciplinary team must aggressively address each of these residents in relation to their fall risk. Implementation of a comprehensive plan of care, maintaining good communication between facility and the patients' physicians and families, and quality charting by the nursing staff, are some of the key elements in preventing litigation. PMID- 12741053 TI - Keep it simple: liberalization of therapeutic diets for older adults. PMID- 12741054 TI - Integrative care--subtle body energies. PMID- 12741055 TI - [Obtaining and analysis of intergeneric somatic hybrids between Brassica napus and "albino" line of Orychophragmus violaceus]. AB - The Orychophragmus violaceus chlorophylldefective line of "albino" type has been obtained by spectinomycin treatment. Somatic hybridization between Orychophragmus violaceus and Brassica napus was performed by fusion of green mesophyll protoplasts of rape and callus protoplasts of the O. violaceus "albino" line. Near two hundred of regenerant plants were selected according to the regeneration type and ability to become green, and were determined as hybrids. Chloroplast DNA in selected hybrids was identical to rape chlDNA, which was confirmed by the PCR RFLP analysis of plastid DNA fragments. Fragments of hybrid mitochondrial DNA analyzed by the PCR-RFLP analysis were identical to fragments of O. violaceus. The nuclear genome of the majority of hybrids was represented by the O. violaceus genome, which was demonstrated by analyses of isoenzymes, DNA telomeric sequences, ribosomal and satellite DNAs, and the RAPD analysis. The cytogenetic analysis of a number of lines has shown variability in the number of chromosomes in the obtained lines. PMID- 12741056 TI - [Hybrids of Aegilops cylindrica Host with Triticum durum Desf. and T. aestivum L]. AB - The hybrids of durum and bread wheat with Ae. cylindrica have been obtained without using an embryo rescue technique. The hybrid output (of pollinated flower number) in the field conditions scored 1.0, 15.3 and 10.0% in the crosses T. durum x Ae. cylindrica, Ae. cylindrica x T. durum and T. aestivum x Ae. cylindrica, respectively. A high level of meiotic chromosome pairing between homologous D genomes of bread wheat and Aegilops has been revealed (c = 80.0 83.7%). The possibility of homoeological pairing between wheat and Ae. cylindrica chromosomes has been shown. Herewith, the correlation between the levels of homological and homoeological pairing is absent. The possibilities of genetic material interchange, including between the tetraploid species, as well as the using of Ae. cylindrica cytoplasm for durum wheat breeding are discussed. PMID- 12741057 TI - [Restriction variability of amplified chloroplast DNA fragments in genus Nicotiana]. AB - Restriction analysis of amplificated chloroplast DNA fragments has allowed us to determine the source of cytoplasm for several natural diploids and amphidiploids of the genus Nicotiana. N. corymbosa or species of Trygonophyllae section donated cytoplasm to N. bigelovii and N. clevelandii, while N. arentsii inherited cytoplasm from N. undulata during the evolution of the genus. Plastomes of Repandae and Nudicaules species orginated from now extinct species of Trygonophyllae section. PMID- 12741058 TI - [Characters of amino acid substitution in LDH-A and LDH-B in the evolutionary line of vertebrates]. AB - Structure of lactate dehydrogenase LDH-A (muscle) and LDH-B (heart) subunits is compared in the evolutionary line of vertebrates from Chondrosteous fishes to Mammals. It is revealed persistent differences between them in the amino acid set determining the physical and chemical characteristics of macromolecules. The polypeptide chain of LDH-A is shorter then that of LDH-B but it contains amino acids with higher molecular weight. In LDH-A polarized amino acids are less in number but charged amino acids are more numerous, positive charged amino acids prevail over negative charged ones. The features of polypeptide structure are discussed in connection with differences in the level of intraspecific variability of allozymes in the evolutionary line of vertebrates. PMID- 12741059 TI - [Geographical variability and subpopulational genetic organization of oak leafroller moth]. AB - Geographically distant populations of oak leafroller moth (Tortrix viridana L.) were characterized by the frequency uniformity for most often alleles and differentiation for rare ones in the esterase and protease loci. Opposite tendency was observed at the subpopulational level. It was supposed that the distinctive spatial distribution of the often and rare alleles was due to different sensitivity of the given groups to the factors of a dynamics. Natural selection and stochastic processes play a major role in the dynamics of often and rare alleles, respectively. PMID- 12741060 TI - [Karyotype of blue honeysuckle species (Lonicera subset. Caeruleae, Caprifoliaceae]. AB - For the first time the karyotypes of diploid (2n = 2x = 18) and tetraploid (2n = 4x = 36) species of Lonicera from the Caeruleae subsection: L. altaica Pall., L. boczkarnikowii Plekh. (L. regeliana Boczkarn.), L. edulis Turcz. ex Freyn (2x, 4x), L. emphyllocalyx Maxim., L. iliensis Pojark., L. kamtschatica Pojark., L. pallasii Ledeb., L. stenantha Pojark., L. turczaninowii Pojark., L. villosa (2x, 4x) (Michx.) Muhl. are described. The species karyotypes from 23 natural populations have shown the considerable generic resemblance that expressed in the similar chromosome morphology and variation range of their length from 1 to 3 microns. The species with the same level of ploidy had the same karyotype formula: 2m + 6sm + 1st in diploids and 4m + 11sm + 3st in tetraploids, respectively. The amphiploid origin of the tetraploid Lonicera species has been shown. Diploid and tetraploid forms of L. edulis and L. villosa were the particular karyotypes but not the 2x and 4x races of the same species, respectively. Specific differences were revealed in the total chromosome length in the haploid set and in the number of satellites and secondary constrictions. Generic resemblance and specific peculiarities of Lonicera karyotypes indicate a common center of the blue honeysuckle origin and a common initial population of karyotypes which evolved into several phylogenetic branches of the Caeruleae subsection: the Central Asiatic--L. iliensis and L. stenantha; the Siberian--L. altaica, L. edulis, and L. pallasii; the Beringian--L. emphyllocalyx, L. kamtschatica, and L. villosa; the Manchurian--L. boczkarnikowii (L. regeliana), and L. turczaninowii. PMID- 12741061 TI - [Genetic structure of unisex hybrid population of frogs Rana esculenta complex in lowland Transcarpathian region]. AB - The biochemical genetic marking and analysis of the ploidy and sexual structure of the green frog populations from the Transcarpathian lowlands have demonstrated that this region is inhabited by the unisexual populations composed of the allodiploid females only. Their genome includes a small portion of the marsh frog genic diversity. This phenomenon is discussed in relation to the unisexual population reproduction problem. The assumption is proposed that in the Transcarpathian hybrid populations the hybrids are spawned by the parthenogenesis. PMID- 12741062 TI - [Biological effect of low doses of alkylating agents in drosophila studies]. AB - The low dose (0.05-0.1 mM) influence of alkylating agents on germ cell survival and male fertility, the level of embryonic and postembryonic lethality as well as the sex-linked recessive lethal (SLRL) frequency induced by high alkylating agent doses was studied in Drosophila melanogaster. The pretreatment of adult males with low doses of methyl and ethyl methanesulfonate (MMS and EMS) did not change or even enhanced EMS cytotoxicity and mutagenicity in both mature sperm and premeiotic cells. On the contrary, the low EMS dose pretreatment of larvae protected them against higher mutagen doses increasing male fertility, decreasing embryonic and postembryonic lethality in F1, and leading to three-fold reduction in the SLRL frequency in F2. The adaptive response was dependent on the Drosophila developmental stage exposed to challenge mutagen doses, since the protection was maximal in larvae and practically absent when the high dose was administered to adult males. The adaptive response observed does not seem to be associated with DNA repair, but it is rather due to other protective mechanisms. PMID- 12741063 TI - [Electron microscopy and ultracytochemistry of blood lymphocytes containing Gall bodies in healthy individuals]. AB - The authors have conducted electron microscopic and ultracytochemical studies of blood lymphocytes of healthy individuals containing Gall bodies (GB) which are typical for CD4+ cell subpopulation. It has been established that GB have quite a complex submicroscopic structure and together with its derivates, granules satellites, as well as mitochondria, microfilaments, rough endoplasmic reticulum canaliculi, and Golgi complex it forms a unique active functional complex. GB show a high activity of acid phosphatase and positive reaction to glycogen. The data obtained suggest the leading role of GB in the production of cytokins and other biologically active substances. PMID- 12741064 TI - [ISSR-PCR in differentiation of gene pools of cattle breeds]. AB - The genetic structure of five cattle breeds from Ukraine was investigated using ISSR-PCR method. Breeding- and sex-specific spectra of amplification products were revealed with two threenucleotide microsatellite primers. The possibility of using ISSR-PCR markers for characterization of genetic relations between breeds of cattle were evaluated. PMID- 12741065 TI - [Visualisation of cyanobacteria in aqueous solution by atomic force microscopy]. AB - Methodical approaches for studying of living cells in aqueous solutions by atomic force microscopy (AFM) are demonstrated. Images of intact cyanobacteria Synechocystis PCC 6803 in TES buffer were captured in tapping mode using aminomodified mica as AFM substrate. Modification of freshly cleaved mica has been done in 3-aminopropyltri-ethoxysilane vapours. The average size of cyanobacteria was determined from AFM images. The linear size of Synechocystis PCC 6803 in TES buffer was equal to 70 x 90 nm and their height was about 20 nm. Possible causes of insufficiently high resolution of the cyanobacteria AFM images in aqueous solutions and possible ways for gaining molecular resolution in studies of structural, functional and micromechanical properties of living cells are discussed. PMID- 12741066 TI - [Plant rDNA: organisation, evolution, and using]. AB - Ribosomal DNA comprises a considerable part of a plant genome and is organized in tandemly arranged repeats composed of conservative coding sequences for ribosomal RNA and rapidly evolving spacer elements. We determined the nucleotide sequences of intergenic spacer regions (IGS) for five species from Solanacaea family: Solanum tuberosum, Atropa belladonna, Nicotiana tabacum, N. tomentosiformis, and N. sylvestris. The detailed comparative analysis of these and some other rDNA sequences allowed us to reveal the general regularities of evolution and functional organization of the rDNA spacer region and to clarify better phylogenetic relationships between the species within Solanacea family. A large body of experimental data on the application of rDNA in plant breeding, taxonomical studies and biotechnology are provided and discussed. PMID- 12741067 TI - Courageous attitude. PMID- 12741069 TI - My experience as a hospital patient. PMID- 12741070 TI - Preeclampsia: theories and speculations. PMID- 12741071 TI - [ALLHAT: Old Hat Hits All? On selection of the optimal antihypertensive drug]. PMID- 12741072 TI - [MADIT II--or how seriously do we take primary prevention of sudden cardiac death?]. PMID- 12741074 TI - Testicle replacement by a brandy bottle--self mutilation. PMID- 12741073 TI - Nicotine dependence: development, mechanisms, individual differences and links to possible neurophysiological correlates. AB - There is now little doubt that the majority of people who smoke tobacco do so to experience the psychopharmacological properties of the nicotine present in the smoke and that a significant proportion of habitual tobacco users become addicted to the drug nicotine. In the US some 80% and in Europe (Germany) 39% of smokers have been classified as dependent according to the diagnostic guidelines of the American Psychiatric Association. As a result, direct nicotine replacement is used increasingly by many people who want to stop smoking. The objectives of this review are to outline the mechanisms involved in the development and maintenance of nicotine dependence and to link behavioural observations to possible neurophysiologic correlates. PMID- 12741075 TI - Elevated serum concentrations of androgens in women with pregnancy-induced hypertension. AB - Alterations of steroid hormone profiles have been suggested to be involved in the pathophysiology of pregnancy-induced hypertension (PIH). The aim of our study was first to investigate serum concentrations of testosterone, dihydrotestosterone, androstenedione and dehydroepiandrostenedione sulfate in women with PIH and normotensive pregnant women and secondly to evaluate an association between elevated serum concentrations of androgens and the development of severe disease. Serum concentrations of androgens were measured in 40 patients with PIH and 40 normotensive pregnant women, matched for gestational age, determined by enzyme linked immunosorbent assay. Multivariate logistic regression models were used to analyze the influence of elevated serum concentrations of androgens on the occurrence of PIH and the development of severe disease. The median serum concentrations of androstenedione and testosterone were significantly elevated in women with PIH compared to controls (6.3 and 5.0 ng/ml, 1.8 and 1.1 ng/ml, p = 0.005 and p = 0.04, respectively). The difference between the median serum concentrations of dihydrotestosterone and dehydroepiandrostenedione sulfate in women with PIH and controls was not significant. Elevated serum concentrations of androstenedione revealed a significant influence on the odds of presenting with PIH (p = 0.043) and were significantly associated with the development of severe disease (p = 0.014). Women with PIH have elevated serum concentrations of androstenedione and testosterone. Moreover, elevated serum concentrations of androstenedione are associated with development of severe disease. PMID- 12741076 TI - [The impact of the Multicenter Automatic Defibrillator Implantation Trial II in a university hospital--do all patients with myocardial infarction and reduced left ventricular function need an implantable cardioverter-defibrillator?]. AB - BACKGROUND AND OBJECTIVES: Patients with coronary artery disease (CAD) and severely compromised left ventricular ejection fraction are at high risk to die from sudden cardiac death. The Multicenter Automatic Defibrillator implantation Trial II (MADIT II) shows a significant benefit of a cardioverter-defibrillator (ICD) therapy compared to standard treatment alone in this selected group of patients. The objective of the present study was to investigate the number of patients who will fulfil the MADIT II criteria and are candidates for prophylactic ICD implantation. METHODS AND MATERIAL: From January to December 2001 a total of 2653 patients underwent coronary angiography at our institution due to angina pectoris, positive exercise stress testing, pathological SPECT myocardial perfusion images, suspected dilated cardiomyopathy or ventricular arrhythmias. According to the MADIT II inclusion criteria patients with significant coronary artery disease (diameter stenosis > 50%), ejection fraction < 0.31% and previous myocardial infarction were included. Exclusion criteria were acute coronary syndromes, patients with ventricular tachyarrhythmias or an existing indication for ICD therapy, and patients with coronary artery bypass graft surgery or percutaneous transluminal coronary angioplasty within the past or following three months. RESULTS: Out of 2653 patients 185 (7%) had an ejection fraction less than 0.31, 149 (5.6%) showed significant coronary artery stenosis and 70 (2.6%) patients fulfilled the MADIT II criteria. The mean age of these patients was 68 +/- 9 years, the left ventricular ejection fraction 24 +/- 6. In 37 patients an ICD system was implanted according to the existing guidelines. 70 patients met the MADIT II inclusion criteria, resulting in an increase of 189% of ICD implantations per year. CONCLUSION: 2.6% out of 2653 patients who were referred to coronary angiography fulfilled the criteria of MADIT II. The expanding indication for ICD therapy will result in an annual increase of 70 (189%) prophylactic ICD implantations in our study population. PMID- 12741077 TI - Are dyspeptic symptoms linked to Helicobacter pylori? A prospective cohort study among medical students. AB - OBJECTIVE AND DESIGN: In Austria, the prevalence of Helicobacter pylori (Hp) infection in healthy people with or without symptoms of dyspepsia has not yet been assessed systematically. We conducted a prospective cohort study among medical students during their rotation in internal medicine to determine whether or not dyspeptic symptoms are linked to Hp infection. Symptoms were assessed before testing for Hp so that knowledge of Hp status could not influence symptom reporting behavior. METHODS: From February 2000 to February 2001 we invited 242 medical students to participate in this trial. Abdominal symptoms were assessed by a questionnaire. Dyspeptic symptoms were defined according to the Rome II criteria: heartburn was not considered a dyspeptic symptom. Severity of symptoms was assessed using the Glasgow dyspepsia severity score (GDSS). After determination of symptoms, students underwent a 13C-urea breath test for Hp. RESULTS: Of 242 students (103 male, 139 female, mean age 26 years, range 21-39 years), 23 tested positive for Hp (10%). Of 34 participants reporting dyspeptic symptoms (14%, GDSS-range 1-11), four were Hp positive (12%). Of the 208 asymptomatic students, 19 were Hp positive (9%). Statistical analysis did not show a significant difference in the frequency of Hp infection between asymptomatic students and students reporting dyspeptic symptoms. CONCLUSION: Hp prevalence in Austrian medical students is similar to that in other western countries. There was no significant difference in Hp prevalence between dyspeptic students and asymptomatic students. PMID- 12741078 TI - Serum level of the circulating angiogenesis inhibitor endostatin in patients with hyperthyroidism or hypothyroidism. AB - Serum level of endostatin, a natural angiogenesis inhibitor, was measured in 12 patients with hyperthyroidism and 9 patients with hypothyroidism. Control values were obtained from 12 healthy individuals. Hyperthyroidism was shown to be associated with an increased level of endostatin and hypothyroidism with a decreased endostatin level. There was no correlation of serum endostatin with thyroid hormone levels. Endostatin is a fragment of type XVIII collagen, and it is possible that reported changes are related to the effect of thyroid hormones on connective tissue metabolism. PMID- 12741079 TI - Autoimmune thyroid diseases in patients with chronic fasciolosis. AB - BACKGROUND: The prevalence of autoimmune thyroid diseases and autoantibodies in patients with chronic fasciolosis (CF) was investigated. METHODS: Thyroid function of 32 patients with CF and 72 patients without fasciolosis infection was evaluated biochemically. Thyroid hormones, antithyroglobulin antibodies (TgAb) and antithyroid peroxidase antibodies (TPOAb) were measured. RESULTS: In patients with CF, the prevalences of autoantibodies against thyroid peroxidase and thyroglobulin were 21.9% (7/32) and 15.6% (5/32) respectively. However, thyroid function disorder was identified in only 3 (9.4%) cases. In the control group, TPOAb and TgAb were determined in 2 (2.7%) cases, but no one had thyroid function disorder. The prevalences of thyroid autoantibodies (chi 2: 7.948, p < 0.01) and autoimmune thyroid diseases (chi 2: 4.007, p < 0.05) were significantly higher in the CF group. CONCLUSIONS: The high prevalence of TPOAb and TgAb in CF suggests that patients with CF should be evaluated for autoimmune thyroid disorders at the time of diagnosis and during the follow-up period. PMID- 12741080 TI - Decreased cutaneous vitamin D-synthesis in heavily melanized individuals: a rare cause for pathologic fractures of the hip. AB - Painful pathological fractures of the femoral neck and the subtrochanteric region of the femur are reported in two women originating from India. After exclusion of renal or intestinal causes, laboratory data on bone metabolism, scintigraphic and radiographic examinations were characteristic for the presence of secondary hyperparathyroidism. Based on vitamin deficiency and low calcium absorption, disturbed mineralization of bone and increased osteoclastic resorption have apparently led to osteomalacia and subsequent fracturing. Fracture localization necessitated surgical fixation in one patient; conservative treatment including protected weightbearing was effective in the other women. After supplementation of calcium and vitamin D3, levels of parathyroid hormone and scintigraphic alterations returned to normal in both patients. In these two cases, pathological fractures of the hip could be attributed to the presence of secondary hyperparathyroidism based on decreased cutaneous vitamin D synthesis. PMID- 12741083 TI - [Pharmacology in Austria in the 20th century: in honor of Professor Heribert Konzett on his 90th birthday]. PMID- 12741081 TI - Bcl-2 expression in a primary leiomyosarcoma of the ovary: a case report. AB - BACKGROUND: Primary ovarian leiomyosarcoma is an extremely rare malignant smooth muscle neoplasm with fewer than 30 documented cases worldwide. Immunohistochemically, ovarian leiomyosarcomas are characterized by the expression of alpha-smooth muscle actin (alpha-SMA). Until recently, no report investigated the role of bcl-2 expression in primary ovarian leiomyosarcoma. We report the management and an immunohistochemical analysis of bcl-2 in a patient with primary leiomyosarcoma of the ovary. CASE REPORT: The patient, a 71-year-old woman, presented with a history of lower abdominal pain and a palpable mass in the lower abdomen. Surgical exploration revealed a large left adnexal mass that had developed from the left ovary and infiltrated one part of the omentum adherent to the left adnexa. Microscopically the left adnexal tumor consisted of spindle cells arranged in smooth-muscle bundles and fascicles. The tumor was characterized by high cellularity and nuclear polymorphism, as well as patchy necrosis and large areas of hemorrhage. Mitotic activity exceeded 5 mitoses per 10 high-power fields with many atypical mitoses. The tumor appeared encapsulated, and the capsule was partially disrupted by tumor cells. Immunohistochemically, the tumor showed a strong staining reaction for muscle actin, alpha-SMA and neuron-specific enolase, and also a weak reaction for vimentin. Strong staining of bcl-2 was detected, with more than 80% of sarcoma cells positive. According to the FIGO staging system for ovarian cancer, the tumor was classified as a primary ovarian leiomyosarcoma stage III C, and the patient was treated with an adjuvant chemotherapy regimen consisting of cisplatin and ifosfamide. Two days after the third cycle of cisplatin/ifosfamid she developed an apoplexy spontaneously and chemotherapy was therefore discontinued. The patient remained without evidence of disease until she died as a result of intercurrent pneumonia 14 months after initial surgery. CONCLUSIONS: Beside the routine histological and immunohistochemical characteristics of primary ovarian leiomyosarcoma, strong staining for bcl-2 was detected. In order to understand more about the nature and the behaviour of this highly malignant neoplasm and to be able to improve the treatment, the prognostic value of bcl-2 has to be investigated in additional studies. PMID- 12741084 TI - [Consensus recommentation for supplying corroboration for increased subsidies for families with children and adolescents with chronic respiratory tract diseases]. PMID- 12741085 TI - [Alternative and naturnaturopathy forms of therapy: a discussion contribution]. PMID- 12741086 TI - The quadricuspid aortic valve. PMID- 12741082 TI - A case of primary tularemic pneumonia presenting with necrotizing mediastinal and hilar lymph nodes. AB - Tularemia is an unusual disease caused by the gram-negative coccobacillus Francisella tularensis. The clinical features of the disease depend on the route of inoculation. Ulceroglandular and typhoidal forms have been recognized as occurring in tularemia, however primary or secondary pneumonic infections have also been reported. Symptoms, laboratory markers and radiological features are non-specific in tularemic pneumonia. Diagnosis is made on clinical grounds and evidence of elevated agglutinating antibodies to F. tularensis (> 1:128). We report a case of primary tularemic pneumonia presenting with pulmonary infiltrates and necrotizing mediastinal and hilar lymph nodes in an otherwise healthy subject from a non-endemic area. Diagnosis of tularemia was obtained serologically, and antibiotic therapy with doxycycline and streptomycin resolved symptoms and radiological abnormalities. We suggest that tularemia should be considered in the differential diagnosis of pneumonia with mediastinal and/or hilar lymphadenopathy. PMID- 12741087 TI - [Unnatural death: nursing home physicians and regional coroners]. PMID- 12741088 TI - [Depression among older nursing home patients. A review]. AB - Depression is a common disorder in later life. The prevalence of depression in aged nursing home patients in 36 studies in various countries was reviewed. Results show prevalence rates ranging from 2% to 61%. Average prevalences were calculated for depressive symptoms, minor depression and major depression each. The averages thus found are 43.9% for depressive symptoms, 25.7% for minor depression and 15.5% for major depression. In order to find an explanation for the variation in occurrence of depression in nursing homes, factors that may have influenced the results are described. Both the definition of depression and the kind of instrument used in measuring depression appear to be highly responsible for the variations found. PMID- 12741089 TI - [Natural and unnatural death: assessment of the manner of death by nursing home physicians and notification to the medical examiner]. AB - The assessment of the manner of death by physicians is often a problem due to unclear regulations on this point. By means of a questionnaire we investigated how 40 nursing home physicians assess the manner of death in 14 different hypothetic cases and whether the physicians should report the case to the medical examiner if confronted with similar cases in practice. Some cases of unnatural death would not be reported. Especially in cases of death after a collum fracture the law and medical practice diverge. The reasons to report or not to the medical examiner were also investigated. Death fitting in the 'descending line of life' was most mentioned. PMID- 12741090 TI - [The legal position of psycho-geriatric patients in nursing homes and residential homes]. AB - Recently the Law on Formal Admission to Psychiatric Hospitals was evaluated for the second time. This law aims to protect the legal position of psychiatric patients, who have been involuntarily admitted. This patient group includes psycho-geriatric patients who are admitted to residential and nursing homes for which this is indicated. In this article the part of the evaluation concerning the internal legal position of psycho-geriatric patients is reported. This part of the study investigated both the knowledge of the law in practice and its correct application, as well as the suitability of the law for this domain of health care. The conclusion can be drawn that residential and nursing homes are aware of the laws purposes, but health care practitioners find it hard to follow certain aspects of the law's procedures, especially regarding the use of restraints. The main reason for this is that the Formal Admission Law was designed for a psychiatric setting, which cannot easily be translated to the position of psycho-geriatric patients. The suggestion is made that the Government will develop new legislation, which reflects special characteristics of psycho geriatric care. Meanwhile health institutions have to take measures in order to improve the legal position of patients in this sector. PMID- 12741091 TI - [Neuropsychiatric inventory questionnaire (NPI-Q): A validity study of the Dutch form]. AB - The Neuropsychiatric Inventory is a clinical instrument for assessing behavioral and psychological symptoms in dementia. It is based on an interview with the primary caregiver. A brief questionnaire form of the NPI was developed, intended for use in routine clinical practice (NPI-Q). This study evaluates the validity of the NPI-Q Dutch form. The NPI-Q was compared to other questionnaires, i.e. the Revised Memory and Behavioural Problems Checklist (RMBPC), the short form Informant Questionnaire on Cognitive Decline in the Elderly (IQCODE-N), the 15 items Geriatric Depression Scale (GDS-15) and the Cognitive Screening Test (CST 20). A data set was used of geriatric outpatients who were referred for neuropsychological assessment (n = 29). Correlations between the NPI-Q and RMBPC Depression- and Disinhibition subscales were relatively high. No relationship was found between the NPI-Q and the RMBPC Memory related behavioral changes subscale, or between the NPI-Q and the IQCODE-N and the CST-20. Informant ratings on the NPI-Q depression-item were related with patient ratings on the GDS-15, especially when patients were relatively mildly cognitive impaired. Caregiver distress was strongly associated with NPI-Q symptom assessment. IN CONCLUSION: our preliminary results support validity of the NPI-Q Dutch form. It is a practical rating scale for assessing neuropsychiatric symptoms in dementia and associated caregiver distress. PMID- 12741092 TI - [Clinical, hematological, biochemical and endocrinological aspects of 32 dogs with hypothyroidism]. AB - During the years of 1996-2001, hypothyroidism was diagnosed at the clinic for small animal internal medicine, University of Zurich, in 32 dogs. Most of the dogs were large breeds. The most frequent clinical characteristics observed were exercise intolerance, obesity, dermatological, neurological and gastrointestinal signs. Predominant laboratory abnormalities were a low red blood cell count, increased concentration of cholesterol, triglycerides and fructosamin. 29 dogs had a T4 below the reference range (< 1.5 micrograms/dl), one dog had a T4 at the lower limit thereof (1.6 micrograms/dl). One dog had a T4 within the reference range (3.4 micrograms/dl), another had a very high T4 of 206.8 micrograms/dl; the results of the latter 2 dogs were interpreted as incorrectly increased T4 values due to in vitro interference with T4-autoantibodies. Diagnosis was confirmed in all of the dogs based on TSH-stimulation testing. Endogenous TSH (cTSH) measured parallelly, was elevated in only 60% of the dogs. In about 67% of the dogs, hypothyroidism was associated with thyroglobulin-autoantibodies. Canine hypothyroidism is a rather rare endocrine disorder in Switzerland. The TSH stimulation test remains the gold standard in confirming the disease; a definitive diagnosis can be challenging for practitioners because bovine TSH, used for the TSH-stimulation test is not licensed for use in dogs. Since assessment of cTSH using current assays shows normal values in a high percentage of hypothyroid dogs, the diagnostic value is only limited. In most of the hypothyroid dogs T4 is decreased, with the presence of autoantibodies to T4, it can be normal or increased. PMID- 12741093 TI - [Retrospective study of the fertility outlook in cows after caesarean section]. AB - Evaluating the possibility of performing a caesarean section on a cow with dystocia, the economic value of the animal usually plays a central role. The aim of this retrospective study was to analyse and to determine possible factors influencing the fertility. In 1998, 1999 and the first half-year of 2000 a total of 207 cows had a caesarean section. 68% of the inseminated cows which gave birth to a living calf became pregnant again, whereas only 46% of the cows with a dead calf were successfully reinseminated. Thus, 59% of all inseminated cows became pregnant, 2.5% of them had an abortion. The occurrence of retained fetal membranes or of endometritis has been shown to have negative effects on fertility. Only 30% of subsequent inseminations were successful in cows which had suffered from endometritis. The probability of being affected by endometritis was twice as high after birth of a dead calf than after a living calf. PMID- 12741094 TI - [Radiographic findings in 4 cows with traumatic reticuloperitonitis caused by a nonmagnetic copper wire]. AB - The goal of this study was to describe the findings in four cows with non magnetic reticular foreign bodies composed of copper. The cows were referred to our clinic because of reduced appetite and a marked decrease in milk production. Based on the clinical findings, a tentative diagnosis of traumatic reticuloperitonitis was made in all cows. The reticulum of all cows was then examined ultrasonographically and radiographically. In all cows, radiographs of the reticulum showed wire-shaped foreign bodies, ranging from 3 to 7 cm in length, which appeared to have penetrated the reticular wall. Two cows (No. 3, 4) had a magnet in the reticulum close to the foreign body but there was no direct contact between the two. A magnet was administered to cows No. 1 and 2, and radiography of the reticulum was performed for a second time the following day. The magnets were observed in the reticulum; however, they did not contact the foreign bodies. Because all the magnets were correctly placed in the reticulum yet, despite close proximity, did not contact the foreign bodies, the latter were thought to be non-magnetic. Cow No. 1 was slaughtered. Left flank laparoruminotomy was performed in the remaining three cows. In all cows, copper foreign bodies ranging in length from 3.0 to 7.0 cm, were found in the reticulum. They had penetrated the reticular wall and were not attached to magnets. The radiographic findings described in the present study are strongly indicative of a non-magnetic foreign body. Ruminotomy is the treatment of choice but slaughter may also be considered. PMID- 12741095 TI - [Prevalence of Fasciola hepatica and Dicrocoelium dendriticum in the cow: inspection in an east Switzerland abattoir]. AB - On the basis of meat inspection in an abattoir in the eastern part of Switzerland, the incidence of liverflukes in cattle was recorded weekly during one year. The prevalence was compared with surveys from the seventies and nineties. The average incidence of Dicrocoelium dendriticum infection did not change. Although the prevalence of Fasciola hepatica apparently slightly decreased, an actual decrease of liverflukes could not be demonstrated. PMID- 12741096 TI - [Traumatic rupture of the common bile duct in a dog]. AB - Extrahepatic biliary tract rupture only rarely occur secondary to blunt or sharp trauma to the abdomen. Clinical symptoms result from chemical peritonitis and are unspecific. Recognition most often is delayed from several days to weeks, when patients suffer from ascites, icterus, dehydration and apathy. Surgical therapy depends of the site of laceration. In the present case, a dog with bile peritonitis was presented two weeks after being hit by a car. Explorative celiotomy revealed a rupture of the common bile duct. A cholecystojejunostomy was performed. PMID- 12741097 TI - [What is your diagnosis? Active schistosomiasis of the bladder caused by Schistosoma haematobium (bilharziasis)]. PMID- 12741098 TI - [Asthma education and self-management: inevitable requirements to enhance disease control and achieve the goals for long-term asthma management]. AB - The patient should be the primary manager of chronic disease guided and coached by a doctor and healthcare workers. The challenge now is to recognise that establishing patient-provider partnerships is an essential step in helping patients to learn and manage chronic illnesses like asthma. This article focuses on what is known about asthma patient education and on reducing the barriers to effectively applying what is known. Whereas traditional patient education offers information and technical skills, self-management education teaches problem solving skills. A central concept in self-management is self-efficacy, the confidence to carry out a behavior necessary to reach a desired goal (outcome efficacy). Effective teaching on chronic diseases must be based more closely on the findings of behavioural research. Therefore, self-management education has to become an integral part of high-quality care in asthma. PMID- 12741099 TI - Antibiotic-associated diarrhea: incidence, risk factors of antibiotics and patients, pathophysiology and differential diagnosis--an interdisciplinary approach to a common problem. AB - Antibiotic-associated diarrhea (AAD) is a common complication of antibiotic treatment, most often seen in non-hospitalised patients. In principle, such diarrhea can be triggered by any antibiotic. An interdisciplinary working group discussed the different aspects of AAD in view of its gastroenterological, microbiological, paediatric, general medical and pharmaceutical implications, also in consideration of the position of patients and health insurance funds. The incidence, risk factors of antibiotics and patients, the pathophysiology of the various types of AAD and the differential diagnosis are reviewed. PMID- 12741101 TI - [A 45-year-old patient from Italy (Calabria) with pain in the shoulder girdle, neck, lumbar and hip areas as well as in both knees lasting for many years]. PMID- 12741100 TI - [Cerebral metastasis in choriocarcinoma a case report]. AB - Choriocarcinoma are malignant neoplastic tumors from the trophoblastic tissue with a tendency to early metastases. Beside pulmonary metastases there are often cerebral metastases, leading to intracerebral hemorrhage often responsible for the first clinical symptoms. In young women, symptoms like vaginal or pulmonary bleeding or neurologic disturbances shortly after a hydatiform mole or a normal pregnancy, accompanied by high levels of HCG in serum and CSF, choriocarcinoma should be considered. Choriocarcinoma are very sensitive to chemotherapy, which consists--depending on the stage of the disease--of a mono- or polychemotherapy. Cure rates are high, even in extended stages with cerebral metastases--as in the case described. Brain metastases with or without oncotic aneurysms can be rapidly controlled by immediate whole brain irradiation. Surgical interventions may be necessary in the case of life threatening bleedings. Levels of HCG in serum and cerebrospinal fluid are good markers to control the effect of therapy. But--as shown in this patient--levels of HCG in CSF may decrease protracted without affecting prognosis. Oncotic aneurysms are rarely reported and mostly detected post mortem. The presented case leads to a more optimistic attitude and demonstrates efficacy of immediately started radio- and chemotherapy. PMID- 12741102 TI - [Abstract of the scientific article--significance, contents, style]. PMID- 12741103 TI - [Stromal stem cells and osteoblastic cellular differon]. AB - The article presents an analysis of the data on the origin, differentiation, morpho-functional characteristics and reactivity of the bone cells taking into account theoretical concepts of histogenetic series and cell differons. The information on the sources and proliferation of cells of the osteogenic line was used as the basis for development of a variant of a scheme describing the differentiation of these cells from a stromal stem cell (SSC) to a bone-lining cell and osteocyte. Considering possible differentiation of SSC into the cells of fibroblastic, chondroblastic, adipocytic and leiomyocytic cell lines, the unified model of mechonocytogenesis is proposed. PMID- 12741104 TI - [Variability of the structure of area 39 of inferior parietal cortex in the left and right hemispheres of adult human brain]. AB - The cytoarchitecture of area 39 of inferior parietal cortex region in the left and right cerebral hemispheres was studied in usual individuals and in professionally gifted world-famed people. The series of frontal sections, stained using Nissl cresyl violet method, were examined. The profile areas of neurons in cortical layers III and V were measured and their distributions were analyzed. In the group of gifted people, individual variability of the characteristics studied was found to be significantly increased, especially in the right cerebral hemisphere, while the relative content of both very small and very large neurons was elevated. PMID- 12741105 TI - [Cytopathological changes in human and animal brain in prion diseases]. AB - The study of brain histological sections of humans and animals afflicted by prion diseases has shown that neuronal vacuolization began in and intensely spread from the distal portions of dendrites. Two types of neuronal cell body death were demonstrated, including cytolysis and pyknosis, the latter being usually associated with dendrite vacuolization. The involvement of major glial types in the pathogenesis of prion diseases was established, which argues against the concept of astrocytosis in favor glyosis. The accumulation of phagocytes (microgliocytes, blood macrophages, pericytes) and emigration of lymphocytes from the blood vessels with diffuse infiltration of brain tissue and formation of aggregates around degenerating neurons, were noted. PMID- 12741106 TI - [Afferent connections of columns of area 17 and 18 of the cerebral cortex formed by neurons of dorsal lateral geniculate bodies in cats]. AB - Horseradish peroxidase (HRP) was injected microiontophoretically into nine single area 17 columns and six area 18 columns of the cat's cerebral cortex. Within the dorsal lateral geniculate nucleus most of the labelled cells were found in the laminae A. The ratio of HRP-labelled cells in lamina A to those in lamina Al (A/A1) was estimated. After injection of HRP in area 17 columns this ratio varied from 0.5 to 2.5, while after HRP injection into area 18 columns it varied from 0.2 to 0.9. These data may indicate that in the areas studied contain the columns that receive afferents from both ipsilateral and contralateral eyes. PMID- 12741107 TI - [Neuronal structure of piriform cortex of anterior amygdaloid body of the rat brain]. AB - The cytoarchitecture and neuronal organization were studied in piriform cortex at rostral and caudal levels of anterior amygdaloid body using the methods of Nissl and Golgi. The division of the piriform cortex into several areas is proposed, including basal area (area basalis), lateral area (area lateralis), and transitional, or terminal area (area terminalis), bordering the insular cortex. It is recommended to subdivide the lateral area into two subareas, L1 and L2. All the areas defined differ in their neuroarchitecture, structure of the layers and neuronal composition. PMID- 12741108 TI - [Organisation of thalamic projections of striopallidum in the dog brain]. AB - The experiments, using the technique of double luminescent labelling, were performed to study the distribution pattern of labelled neurons in thalamic nuclei depending on location of injected luminescent label in functionally similar or functionally diverse areas of striopallidum in the brain of 16 dogs. The peculiarities found in the organization of canine thalamo-striopallidum projection system indicate high level of its specificity, as it was found that not only motor and limbic areas of striopallidum, but also its functionally related areas receive mainly, the separate inputs from different cell groups. The centromedian nucleus was found to contain the groups of diffusely mixed cells, containing double labelling, that innervate functionally different segments of nucleus caudatum. In the centromedian, parafascicular, central medial and mediodorsal thalamic nuclei, projection neurons form similar cell populations, innervating different segments of striopallidum structures, related to the single functional system. The same striopallidum areas receive projections from a small number of neurons by axonal collaterals. PMID- 12741109 TI - [Involvement of actin in electrotonic conductivity in the mixed synapses of goldfish Mauthner neurons]. AB - The effect of highly specific and selective actin-polymerizing and labelling agent, phalloidin, on electrotonic conductivity and structure of the mixed synapses of goldfish Mauthner neurons (MN) was studied. It was shown that the paired subthreshold electrostimulation of afferent input against a background of phalloidin application resulted in the average 80% increase of the amplitude of MN response to the second stimulus. In control group it increased by only 10% and was observed only after suprathreshold stimulation, while subthreshold stimuli were ineffective. We interpret these data as the manifestation of increased conductivity of the mixed synapses, induced by actin polymerization. At the ultrastructural level, phalloidin application at MN and their mixed synapses increased the size and number of actin-containing desmosome-like junctions, as well as the number of fibrillar bridges crossing their cleft. Using the phalloidin-colloid gold marker, the actin nature of these bridges was demonstrated. Interdependent morpho-functional changes found in the mixed synapses, provide the indication of actin involvement in the conduction of electrotonic signal through the mixed synapse. The bridges crossing the cleft of desmosome-like junction could be the structural substrate of this process. PMID- 12741110 TI - [Morpho-functional changes of the spinal cord neurons after epidural administration of lidocaine]. AB - The analysis of morphological and histoezymological changes of spinal cord neurons has been performed in outbred dogs following epidural infusion of isobaric 2% lidocaine solution. In the control group the animals received epidural infusion of 0.9% saline. The results obtained indicate the absence of pathological structural and metabolic changes in the nervous tissue after lidocaine application. Certain signs of morpho-functional reorganization were noted in spinal cord neurons of animals in both experimental and control groups. The registered changes were found to vary within the physiological fluctuation limits and are rather indicative of the functional reaction of the studied structures of the nervous tissue to the epidural injections of either saline and lidocaine. PMID- 12741111 TI - [Cardiomyogenesis in the embryonic period of human development]. AB - The morphological analysis of major processes involved in cardiac myogenesis (including cytodifferentiation, proliferation, integration and apoptosis) was carried out by studying the myocardium of 52 human embryos at weeks 4 to 8 of development. The data provided by this study, permit to draw a conclusion on the coupling of high proliferative activity of cardiomyocytes, their active divergent differentiation, formation of the integrative intercellular communications and manifestations of programmed cell death relations during this period of ontogenesis. Starting with week 6 of intrauterine life, the developing atrial and ventricular cardiac muscular tissue demonstrates the heteromorphy of constituting cardiomyocytes, which differentiate into three types: contractile, contractile secretory and conducting ("light"). PMID- 12741112 TI - [Energy metabolism of blood lymphocytes after insertion of titanium implants]. AB - Blood lymphocyte dehydrogenase activity (SDH, LDH, mitochondrial and cytoplasmic alpha-glycerophosphate dehydrogenase, NADH-diaphorase) was studied in 140 Wistar rats at different time intervals following intraosseous insertion of titanium implants. It was shown that implantation was accompanied with an increase in enzyme activities indicating a rise in the intensity of energy production processes. Cytochemical analysis of lymphocyte enzymes enables to formulate a "prognostic scenario" describing titanium implant interaction with biological environmental factors and organism tissues to assess the long-term results of implantation. PMID- 12741113 TI - [Endocrine cells in the mucosal epithelium of the porcine large intestine]. AB - Using the methods of light and electron microscopy, mucosal epithelium in the colon of adult pigs was studied with special reference to the amount and distribution of endocrine cells. The total amount of endocrine cells tended to increase caudally. Four types of these cells were demonstrated including EC, L, D and D1. The proportion of EC-cells was found to fall from cranial segments of the colon to caudal ones. According to the results of ultrastructural analysis, endocrine cells in the mucosal epithelium of porcine colon, as well as the cells of the adjacent tissues, are in the state of functional exertion. PMID- 12741114 TI - [Differentiation of human pancreatic endocrine cells in the period of fetal development]. AB - The stereological analysis of electron micrographs of pancreatic endocrine cells (A and B) obtained from human fetuses at gestational months 4 to 7 was performed to assess the volume densities of membranous organelles (mitochondria, rough endoplasmic reticulum, Golgi complex), polysomes and secretory granules. The differences were established in the ratios of the volume densities of organelles and secretory granules in the process of differentiation. The morphometric characteristics of total A- and B-cell populations as well as the quantitative data on the structural organization of endocrine cells at various stages of differentiation are presented. PMID- 12741115 TI - [Rat reproductive cells as biological indicators of the effect of environmental factors]. AB - The effect of one-month-long inhalation exposure of male rats to sulphureous anhydrate, phenol, ammonia or to the mixture of these substances with formaldehyde and lead acetate, on the parameters of spermatogenesis was studied. The exposure of animals to sulphureous anhydrate or to the mixture indicated above resulted in the asynchronization of spermatogenic cycle, which was more significant than that caused by the separate application of substances studied. This mixture induced a 20% reduction in the number of tubules, containing spermatogonia, and a 20-30% increase in the number of tubules, containing spermatozoa. The structural and functional study of spermatogenic epithelium, considering stages of spermatogenic cycle, indicated that some cell populations form associations that are targets for the toxic agents studied. PMID- 12741116 TI - [Effect of different somatotypes on the intensity of changes of indices of height and body mass during early childhood]. AB - Anthropometric study of 1425 children of both sexes aged 4-7 years followed by somatodiagnostics according to R. N. Dorokhov method has revealed that at the age of 4 significant proportion of girls belonged to macrosomal type. In the age period of 5-7 years, most part of the children belonged to mesosomal type. However, among the girls of 5-6 years and the boys of 5-7 years the significant proportion belonged to microsomal type (21.1-28.3% of children studied). The growth of height and body mass is more intensive in girls as compared to boys during the whole period of the first childhood. This regularity was found in all the somatotypes with the exclusion of the boys of macrosomal type aged 6-7 years, in whom the maximal reduction of growth of body length was observed. PMID- 12741117 TI - [Changes of body mass components and body constitution in boys 7-17 years of age]. AB - The longitudinal studies of boys aged 7-17 years were performed to trace the dynamics of growth processes, indices of endomorphism, mesomorphism and ectomorphism according to type of body build. The indices of somatotype stability, proposed by the authors, depending on the age and body build, are discussed. It is shown that the least variable are the boys of asthenoid and digestive types of body build, while those belonging to muscular and thoraco muscular types undergo changes most frequently. At the age of 9-10 years the somatotype variability is the greatest. By the age of 15-17 years the proportion of the boys belonging to asthenoid-thoracal type is increased, that is characterized by a deceleration of skeletal muscles development and an increase in the ectomorphism index. PMID- 12741118 TI - [Scar formation in the liver]. AB - The review examines the current data on the cellular and tissue mechanisms of scar formation in the liver. On the basis of the analysis of the literature, authors conclude that to minimize the destructive processes in the liver, the most advisable and promising method during the operations on this organ is the use of laser surgical devices. However, the problems of the selection of the optimal regimes of laser radiation and of the methods of conducting these operations to promote reliable and rapid healing of postoperative wound of the liver. PMID- 12741119 TI - [The principle of feedback in optimization of the method of teaching the course of histology, cytology, and embryology]. PMID- 12741120 TI - [Our teacher (the 100th birthday of Boris Pavlovich Khvatov )]. PMID- 12741121 TI - [How to survive and preserve the departments]. PMID- 12741123 TI - An ontogenetic approach to the assessment of plant resistance to stress factors based on the method of chlorophyll fluorescence induction. PMID- 12741122 TI - Physicochemical properties of artificial bilayer membranes from tissues of vertebrates. PMID- 12741124 TI - Association of the mechanisms of green cell resistance with changes in the parameters of modulated pulse fluorescence under the exposure to atmospheric drought: localization of damage in the link P680QA. PMID- 12741125 TI - The influence of structural phase transition on the temperature dependence of the rate of charge recombination P+QA(-)-->PQA in Rhodobacter sphaeroides reaction centers. PMID- 12741126 TI - Electrodiffusion gradients of ion concentration resulting from self-formation in a flow for electrofocusing of weak electrolytes. PMID- 12741127 TI - Specific effects induced by gamma-radiation on the fine structure of the photosynthetic apparatus: evaluation of the pattern of changes in the high-order derivative spectra of a green leaf in vivo in the red spectral region. PMID- 12741128 TI - Chicken alpha-globin gene cluster is preceded by the nonerythroid-specific gene, the beginning of which is colocated with the replication origin. PMID- 12741129 TI - Regulation of the activity of adenylate cyclase and protein kinase a of the infusorians Dileptus anser and Tetrahymena pyriformis by biogenic amines and peptide hormones. PMID- 12741130 TI - Natural selection in a flow as a universal mechanism of evolution of prebiological autocatalytic systems. PMID- 12741131 TI - Bisalkaloid derivatives of dicarboxylic acids on the basis of lupinine, anabasine, and cytisine as reversible cholinesterase inhibitors. PMID- 12741132 TI - Specific effect induced by subminute amounts of ascorbic acid on the fluctuations of transmission factor of water in the infrared spectral range. PMID- 12741133 TI - Different sensitivities of cholinesterases of the comandor squids (Berryteuthis magister) from different parts of its range to bislupinine derivatives of dicarboxylic acids. PMID- 12741134 TI - Photoaggregation of water-soluble protein (carboanhydrase) induced by the ultraviolet radiation of Xe-Cl laser. PMID- 12741135 TI - Detection of recombinant products during PCR amplification of DNA containing direct alu repeats. PMID- 12741136 TI - Synthetic polymers in studies on the adsorption of viral particles. PMID- 12741137 TI - Genetic instability of (GATA)n microsatellite DNA repeats and somatic mosaicism in the unisexual lizards Darevskia unisexualis. PMID- 12741138 TI - [Microvessel density and endothelial area in assessment of angiogenesis in patients with laryngeal cancer]. AB - The growth of solid tumours and the process of metastasis are dependent on the formation of new blood vessels. Indeed, the intensity of angiogenesis has been shown to be increase in various human tumours. The main aim of this work was the assessment of angiogenesis intensity (AI) in patients with laryngeal cancer using two methods: microvessels density (MVD) and microvessels area (MVA). The group of 151 patients with laryngeal cancer, surgically treated with minimum 5 years observation, was multi-variously analysed. Paraffin--embedded tissue sections from each case were stained with a monoclonal antibody raised against FVIII and CD34 antigens. Intensity of angiogenesis (IA) was measured as microvessels density (MVD) and microvessels area (MVA) by mean of computer image analysis. There were statistically significant correlation between IA measured as MVD using CD34 and tumour size and nodal metastasis. IA described as MVA correlated with: staging, tumour size, nodal recurrences and metastasis. IA measured as MVD after using anti-FVIII correlated with tumour size and nodal metastasis. MVA correlated with staging and nodal recurrences. Overall survival was related to IA measured as MVD using anti-CD34 and anti-FVIII. Disease-free survival was related to IA measured as MVD using anti-VIII. Intensity of angiogenesis has prognostic value in patients with laryngeal cancer. Both methods: microvessels density and microvessels area seems to have similar value in assessment of angiogenesis intensity. PMID- 12741139 TI - Atypical mycobacterial tuberculosis--a diagnostic and therapeutic dilemma? Case reports and review of the literature. AB - In immunocompetent preschool children cervical lymphadenitis is a common clinical presentation of atypical mycobacteria. Its rapid diagnosis and treatment is still a challenge, because accurate diagnostic procedures for atypical mycobacteria are still not yet available in routine practice. Two children suffered from craniojugular (16 months old girl) and infraauricular (2.5 years old boy) located neck masses which showed resistance to the medical treatment. In the first case an abscess splitting took place initially, followed by an anti-tubercular drug treatment and necessary surgical reintervention. In the second case surgical removal of all involved lymph nodes, infiltrated surrounding soft tissue and involved skin areas were followed by medical treatment. In both cases presumed infection with mycobacterium tuberculosis was not confirmed, but atypical mycobacteria could be isolated both. In the first case atypical mycobacterium could be specified as mycobacterium avium complex and in the second case as mycobacterium malmoense. Both bacilli showed sensitivity towards medical treatment with clarithromycin, whereby in one case only the surgical reintervention led to a complete removal of clinical symptomatic. In cases of presumed tuberculous neck lymph node infections differential diagnosis of an atypical mycobacterial lymphadenitis should always be supposed, because medical and surgical treatment differ fundamentally. PMID- 12741140 TI - [Evaluation of advanced laryngeal cancer extension in clinical and pathomorphologic examination]. AB - The method of choice in larynx cancer treatment is surgery. The radical surgical procedure must be based on the proper assessment of loco-regional tumor extenstion. The aim of the study was the estimation and comparison of immensity of larynx cancer during clinical, pathological and microscopic examination, as well as the defining of anatomical and functional conditions which impede laryngoscopy. 180 patients with larynx cancer treated surgically in the Department of Otolaryngology of the University School of Medical Sciences in Poznan and in the Otolaryngology Ward in Pila. In group of 123 patients total laryngectomy and in 57 patients partial laryngectomy was performed. To conduct the analysis of discrepancies between cT and pT as well as cT and mT assessment "big" and "small" immensity features were used. The big immensity was thought to be two-grade difference of locoregional advancement, the small one-one-grade. Discrepancies between cT, pT and mT were stated in transglottic tumors the most often and in multifocal-rarely. The biggest differences in cT, pT and mT assessment occurred in supraglottic tumor localisation. The high correlation between pT and mT local tumor extension assessment was observed. PMID- 12741141 TI - [Pathomorphology of the thyroid gland after treatment of laryngeal cancer]. AB - The aim of the report was the explanation of the pathogenesis of thyroid gland dysfunction after laryngeal cancer treatment. Assessment of histopathological specimens from thyroid gland was done. They were taken from the following patients: a) irradiated primary with the radical dose (n = 5), b) after combined therapy (n = 2) and c) at the autopsy from the persons aged between 30-80, without thyroid gland disease history (n = 30). It was found that the thyroid gland underwent degenerative changes after external irradiation or combined therapy, which became more intense with the passage of time. They consist among other things in diminishing of the follicle lining cells with the obliteration of the follicle structure of thyroid, damage to the blood vessel walls, fibrotic tissue proliferation with the following gland atrophy. In the histological specimens of the thyroid gland taken at the autopsy distinct obliteration of the follicle structure with the atrophy of follicle and intensified fibrotic process after 80 years of age was seen. Results of the investigations suggest that the main pathogenic factor in hypotyhyreosis after radiotherapy or combined therapy are the degenerative changes in the thyroid gland and their overlapping with circulatory disorders after surgery. Morphological changes in thyroid gland following aging increase the risk of hypotyhyreosis. PMID- 12741142 TI - [Activity of the enzymatic antioxidant system depends on the clinical stage of laryngeal cancer ]. AB - The goal of this study was to examine if there is any connection between enzymatic antioxidant system and larynx cancer staging. The levels of superoxide dysmutase, catalase and glutathione peroxidase, 6-phosphogluconic dehydrogenase and glucoso-6-phosphate dehydrogenase were measured in blood of 28 larynx cancer patients. The results show that the activity of this system is lower in sick patients than in healthy control group. It was becoming lower group of patients with more advanced cancer. PMID- 12741143 TI - [Prognostic significance of sinus histiocytosis and lymphocyte predominance in lymph nodes of patients with laryngeal carcinoma]. AB - The prognostic value of morphological parameters such as sinus histiocytosis and nonspecific lymphocyte predominance in removed neck lymph nodes in patients with squamous cell carcinoma of the larynx was presented. Long term results of the treatment in group of 179 patients have shown that regional lymph node reactivity (sinus histiocytosis and reactive lymphadenitis) is a good prognostic factor, while lack of stimulation is a factor of bad prognosis. PMID- 12741144 TI - [Glandular neoplasms of the external auditory canal--clinical and morphologic observations]. AB - In the article cases of glandular neoplasms of the external auditory canal were presented. Ceruminous adenoma was diagnosed in three persons, adenoid cystic carcinoma--in three patients and ceruminous adenocarcinoma in one patient. All of them were treated surgically or by combined method with radiotherapy. Difficulties with exact histopathological diagnosis and with choice of a method of treatment were stressed. Classification of the glandular neoplasms of the external auditory canal was also presented underlying a histopathological and clinical diversity of these tumors, which had previously been incorrectly qualified in one group ceruminoma. PMID- 12741145 TI - [Methods of closure for pharyngocutaneous fistula after laryngectomy]. AB - The majority of salivary fistulas after laryngectomy or after laryngo pharyngectomy close spontaneously within 2-3 weeks, and the remaining require surgical reconstruction. In the years 1987-2000 the pharyngo-cutaneous fistula was closed in 19 patients. The fistula developed after laryngectomy because of larynx cancer T3 and T4. 13 patients were irradiated before. In all the patients laryngectomy was combined with unilateral or bilateral neck dissection. Salivary fistula was closed in one stage operation using pectoralis major myocutaneous flap: 1) skin island of the flap to replace defect of the neck skin and mobilized mucosa from inside, 2) skin island for lining and deltopectoral skin flap from outside, 3) skin island for lining and free split skin flap for external cover, 4) skin island of the flap divided in two paddeles, one for lining and another for external cover. Indications for each method were discussed. Very good result of total fistula closure was obtained in 15 out of 19 patients. The fistula occurred again in 4 patients, in two of them the failure was caused by cancer recurrence. PMID- 12741146 TI - [Sinus tympani in surgery of the middle ear and skull base]. AB - Tympanic sinus is located at the medial wall of the tympanic cavity. It's visualisation is difficult from most of approaches. Tympanic sinus is often filled with cholesteatoma in course of chronic otitis. Intraoperative assessment of tympanic sinus was performed in 29 patients with facial recess cholesteatoma, and in 20 patients with acoustic neuroma operated with translabyrinthine approach. In the group of patients with cholesteatoma four patients underwent canal wall up tympanoplasty with posterior tympanotomy, 25 underwent canal wall down tympanoplasty. Tympanic sinus was assessed during surgery with 30 degrees endoscope. In all cases depth of the sinus was measured with the use of calibrated hook. In 11 patients depth of the sinus exceeded 4 mm. In 6 among these features of bone destruction was found. PMID- 12741147 TI - [Nasal provocation tests with house dust mite in patients with perennial rhinitis]. AB - The aim of this study was to evaluate a response of the upper airways mucosae to allergens house dust mite: Dermatophagoides farinae and Dermatophagoides pteronyssinus. Compared were results of nasal challenge with results of skin prick tests. 87 patients with perennial rhinitis allergic to Dermatophagoides farinae and/or Dermatophagoides pteronyssinus were included. There were 58 men, 29 women, age 18-54 years, mean age 26 years. Nasal provocation tests were performed first with solvent, and next if flow rate did not change more than 20%, with Dermatophagoides farinae and Dermatophagoides pteronyssinus extracts. The nasal response was evaluated with active anterior rhinomanometry. Nasal flow was measured at 15, 30 and 45 minutes after allergen challenge. There were 99% positive skin prick tests Dermatophagoides farinae and 94% positive skin prick tests Dermatopagoides pteronyssinus. Nasal provocation tests with Dermatophagoides farinae extract were positive in 63% patients and Dermatophagoides pteronyssinus extract--in 62% patients. Majority of nasal challenges were positive at 15th minute. There is association between skin prick tests Dermatophagoides farinae and Dermatophagoide pteronyssinus 4+ and positive nasal provocation tests with the same allergen (p < 0.05). We have found correlation between skin tests Dermatophagoides farinae and Dermatophagoides pteronyssinus (correlation index = 0.58) and no correlation between nasal provocation Dermatophgoides farinae and Dermatophagoides pteronyssinus extracts. CONCLUSION: Nasal provocation tests are more specific than skin prick tests in patients with perennial rhinitis. PMID- 12741154 TI - [Distinctness and discrimination of phonemes in children with cochlear implants- preliminary report]. AB - There are many tests prepared for speech evaluation for different age groups and with any disabilities. The percent of correct repeated mono- or bisyllabes words create the final result. Sometimes this score is lower than real oral communication skill of CI patient. In the years 1994-2002 in Poznan 158 cochlear implantations were performed. Since few months the evaluation of the distinctness and discrimination of phonems of cochlear implants children are being conducted. It was found some differences between distinctness and discrimination of phonems and discrimination of words. It means that scores achieved by children in standarized words--tests do not reflect the real level of speech development of each child. PMID- 12741155 TI - [Two cases of rare unilateral sensorineural hearing loss]. AB - We report a case of unilateral sensorineural hearing loss due to Borrelia burgdorferi infection and in a patient with haemophilia B and hepatitis C treated with alpha-interferon. An audiological check-up of that kind of patients is indicated in order to exclude hearing loss. PMID- 12741148 TI - [Endoscopic reconstruction of the anterior skull base in cerebrospinal rhinorrhea]. AB - Cerebrospinal rhinorrhea is a discharge of cerebrospinal fluid caused by the break continuity in dura mater and by bone defect in the base of anterior skull base. The pathological connections appear mainly in the regions of the skull base with the weaker bone structure (the roof of the frontal, ethmoid, sphenoid sinus and cribriform plate). The aim of the study was presenting the possibility of the endoscopic-surgical closing the fistula within the ethmoid roof. The E.N.T Department in Poznan treated 5 patients with cerebrospinal rhinorrhea. In four cases, cerebrospinal rhinorrhea was caused by operative trauma. In one case the rhinorrhea could not be established. All patients with traumatic rhinorrhea underwent operation treatment based on covering of the loss in dura mater with a piece of mucous membrane together with perichondrium of nasal septum or lyophilised dura. The material used for the plastic operation was sealed by tissue adhesive. In 2 cases rhinorrhea recurred. Using the endoscopic technique, after the identification the leak of the cerebrospinal fluid within ethmoid roof, the fistula has been reconstructed with use of the adipose tissue and temporal muscle fascia. Both cases led to complete recovery. Easy access, precision and accuracy of performance the surgery, the approach without external incision of the patient, makes the endoscopic technique very valuable method in treating rhinorrhea caused by the loss in ethmoid roof and cribriform plate. PMID- 12741151 TI - [Multitude of symptoms in Wegener's granulomatosis]. AB - It was introduced the interesting case of Wegener's granulomatosis with full sinus, neurological and ophthalmological symptoms. Entire synthesis of clinical aspects of Wegener's disease was made The course of the disease ascertained superior orbital fissure syndrome, glossopharyngeal paralysis. Disease was concerning on c-ANCA test. During immunosuppressive therapy the patient showed progressive better symptoms, until almost total remission. The diagnosis and treatment of that disease was showed. PMID- 12741152 TI - [Multicenter, open clinical investigation on using cefprozil in therapy of otitis media in children]. AB - We present results of multi center, open clinical trial on using cefprozil (cephalosporin II generation) in acute otitis media in children. Results indicate very good treatments effect (89.3%) and good tolerance of the drug (10.7% side effects). PMID- 12741150 TI - [Evaluation of epithelial proliferation and apoptosis in cholesteatoma of adults]. AB - The aim of the study was to determine the correlation between proliferation and apoptosis in cholesteatoma epithelium with that in normal skin. Cholesteatomas were collected from 20 patients during surgical procedures. 13 normal skin preparations collected from outside of the external ear were used as controls. Dako detective kit with Ki-67 antigen and PCNA was used to assess cell proliferation. Immunohistochemical reaction intensities in the cells were determined. Mebstain Apoptosis kit Direct with antigen APO 2.7 were used to estimate apoptosis. Positive PCNA cells in the suprabasal layer were observed in 9 preparations, 9--in parabasal layer, and in 2--in both basal and stromal layers. Skin preparations showed decreased proliferative ability in comparison to cholesteatoma epithelium. The presence of Ki-67 antigen was presented in cell nuclei of parabasal and stromal layers mainly. Apoptotic cells were detected in suprabasal, prickle and granular cell layers of cholesteatoma epithelium. CONCLUSIONS: 1. Proliferation and apoptosis are significantly higher in cholesteatoma epithelium as compared with the epidermis of the skin. 2. Proliferative activity and apoptosis were observed mainly in the suprabasal layers in the cholesteatoma epithelium. 3. Hyperproliferation and apoptosis in cholesteatoma do not depend on the thickness of the epithelium. PMID- 12741149 TI - [Nonallergic rhinitis with eosinophilia syndrome: state of knowledge]. AB - The aim of our article was to present the actual state of knowledge of the non allergic rhinitis with eosinophilia syndrome (NARES). NARES is characterized by perennial rhinitis and nasal eosinophilia in the absence of allergy in patient's history, negative skin tests and normal IgE. This syndrome was described 20 years ago, but studies on it are scarce. NARES is more frequent in females at mean age. Family history is positive in part of cases. The clinical symptoms are sneezing, watery rhinorrhea, nasal obturation and nasal itching. The precipitating factors are often nonspecific irritants, weather changes or strong odors. It is not sure whether NARES is a form of a vasomotor rhinitis or a first stage of aspirin triad. A presence of bronchial hyperactivity was detected in part of patients with NARES. The crucial feature of the non-allergic rhinitis with eosinophilia syndrome is nasal eosinophilia. Its diagnosing value, mechanisms and different methods of its measurement and differential diagnosis are discussed. The treatment of NARES with nasal steroids is highly effective. The patients may also benefit from the treatment with the oral antihistamines. PMID- 12741153 TI - [Histopathologic changes in hyperplastic palatine tonsils of children with coexisting atopy]. AB - Research done in respect to tonsils on people dealing at the same time with allergy, is seldom the subject of dissertation in polish as well as world wide medical literature. In the enclosed dissertation, writers present the results of histopathological of hyperplastic tonsils, taken from children with coexisting atopy. It is worthwhile to remark, that many of these cases, the presence of eosinophilia and subepithelial oedema was observed. The tonsillar corpuscle have been also described as ingredients in some of the talked about medical samples. PMID- 12741157 TI - [Subperiosteal orbital hematoma as a sinusitis complication]. AB - A case of subperiosteal orbital hematoma as a sinusitis complication--very rarely described in otolaryngological literature--is presented. Taking into account clinical examination and CT there was diagnosed subperiosteal orbital abscess but during surgery there was found hematoma. There has been discussed a clinical picture and a likely pathological mechanism of the complication as well as the performed surgical treatment. PMID- 12741156 TI - [Electrophysiologic tests of the auditory apparatus and vestibular organ in Usher syndrome]. AB - Usher syndrome is a congenital autosomal dominant inherited disease characterised by hearing, balance and vision problems. The aim of the work was an evaluation of hearing and vestibular organ in the chosen group of patients with Usher syndrome diagnosed. Material consisted of 10 persons (5 males and 5 females) aged from 16 to 46 years with Usher syndrome diagnosed. All the patients have been in constant ENT and Ophthalmology Clinics' care. Complex hearing and balance organ, including posturography, as well as complex ophthalmologic examination were performed in each case. Medium or severe perceptive cochlear hearing loss, proper function of vestibular organ and slight elevation of stabilograms parameters were found in 6 cases. In 4 cases there were severe bilateral hearing loss, lack of vestibular function and considerable worsening of stabilograms parameters. In ophthalmologic examination in all the cases there was bilateral symmetrical dystrophy of the retina of various intensification. We concluded that hearing and balance organ examination in Usher syndrome can state accessory diagnostic aspect that might help to distinguish subtypes of the syndrome. In this way audiologic and otoneurologic tests could help to define prognosis of the disease in the individual cases. A necessity of close co-operation of genetics, ophthalmologists and ENT doctors as well as psychologists was stressed. PMID- 12741159 TI - [Examination of the nasopharyngeal cavity in the second part of the XIX century]. AB - The examinations of the nasopharyngeal cavity were initiated in the beginning of the second half of XIX century. Johann N. Czermak (1828-1873), physiologist, Ludwig Tuerck (1810-1868), neurologist, Karl Stoerck, Friedrich Semeleder, Lori, Legrange and others performed exploration of this region. In the second half of XIX century and later palatine hooks, loops, threads of silver tube, pincers, tapes, spatula were used by many rhinolaryngologists. Emanuel Zaufal (1837-1910) was a creator of original method of exploration of nasopharyngeal cavity by pipe "Nasenrachentrichter". Gyergay and Jankauer examined the nasopharyngeal cavity directly. PMID- 12741158 TI - [Granular cell tumor (Abrikosov's tumor) of the larynx--literature review and 2 case reports]. AB - Granular cell tumors are uncommon neoplasms of the head and neck and they are rare in the larynx. About 200 cases had a laryngeal setting. The origin of these tumors is still unknown, but most authors believe it to be neural in origin. Authors described two laryngeal sites of granular cell myoblastoma. In the first case (44 year old man) tumor was located at the vocal fold. It was removed by laryngofissurae. In the second case (56 year old woman) tumor was located at the arytenoid cartilage and was removed radically during Kleinssaser directoscopy. PMID- 12741160 TI - [Significance of genetic factor in pathogenesis of laryngeal squamous cell carcinoma]. PMID- 12741161 TI - [Use of cell lines for analysis of chromosome aberrations during the course of laryngeal squamous cell carcinoma]. PMID- 12741162 TI - [Analysis of physiotherapy outcome in patients with Bell's peripheral facial nerve palsy]. PMID- 12741163 TI - [Late evaluation of indications for and effectiveness of adenoidectomy in children on the basis of selected clinical and audiologic tests]. PMID- 12741165 TI - [Interculturalisation of medical education in the Netherlands]. AB - The number of immigrants within the Netherlands is rapidly increasing. Towards the year 2010 two million first- or second-generation allochthonous persons will be living in the Netherlands. Their origin is extremely diverse. One district of Rotterdam contains more than 100 different nationalities. Medical education should prepare for these changes in the patient population. Education on import diseases as well as on communication via interpreters is necessary. The characteristics of the Dutch healthcare system (partnership between physician and patient, limited use of drug therapy, emphasis on lifestyle advice, and a central role of primary care) should remain the hallmark of medical education. The rapid influx of second-generation, non-western medical students to the medical faculties should be used to realise internal changes to the medical education system, so that it is adapted to the different medical needs of all persons in the Netherlands. PMID- 12741164 TI - [Brain abscess: a difficult diagnosis]. AB - A 40-year-old man had experienced headaches for 6 days and a 51-year-old man (2 weeks after an operation for perianal abscess) had experienced tingling sensations in the left hand for 10 days. After an epileptic seizure both underwent a CT scan of the brain. On these an abnormality was visible, probably a malignant astrocytoma. After several days of complaint reduction with dexamethasone, drowsiness and leftsided hemiparesis occurred. Emergency operations revealed a brain abscess. In the younger patient drainage and the administration of antibiotics were followed by fatal brain oedema. In the eldest drainage and the administration of antibiotics were followed by the extraction of infected teeth; he recovered with a slight loss of strength in the left hand. Brain abscesses are rare in the Netherlands. The diagnosis can be difficult because clinical signs and symptoms are not specific and because an underlying systemic infection is often not apparent. Diffusion-weighted magnetic resonance imaging can nowadays differentiate purulent brain processes from cystic brain tumors. Early treatment (burr hole aspiration and antibiotics) is usually curative. Nevertheless, mortality continues to be almost 10% and (permanent) morbidity 45%. PMID- 12741166 TI - [The value of the pulmonary-artery catheter: not ruled out, but not proven either]. AB - A number of prospective, randomized trials have recently been published on the effects of using the pulmonary artery (Swan-Ganz) catheter in the peri-operative management of high-risk patients and in the treatment of critically ill patients on the intensive-care unit. These studies show that using the pulmonary-artery catheter does not lead to increased survival. It cannot be excluded that more beneficial effects would have been found with other treatment goals, or if different populations had been studied. However, based on the present evidence from the literature, the routine use of the pulmonary-artery catheter can no longer be defended. PMID- 12741169 TI - [Diagnostic image (136). A boy with coughing fits and subconjunctival hemorrhage. Subconjunctival hemorrhage secondary to whooping cough]. AB - An 8-year-old boy suffered from persisting coughing fits for two weeks and subconjunctival haemorrhage since one week. In paired sera serological proof of infection with Bordetella pertussis was found. PMID- 12741167 TI - [Novel approaches; improved diagnosis and therapy with DNA microarrays. I. Technology and data analysis]. AB - With DNA microarrays it will be possible to refine diagnostics and treatment with the use of genome wide information on a patient's sample. Microarrays are currently applied to unravel gene functions, pathogenetic mechanisms, metabolic routes and the effects of drugs on these, to refine diagnostic classifications and prognostic indexes, and to find new targets for therapy. With genotyping it will be possible to generate risk profiles for certain diseases and to assess the likelihood of a given drug-related side effect. Furthermore, it may accelerate research of mutations in genes for which the normal DNA sequences are already known. The basic principle of DNA microarrays is that thousands of different DNA sequences, each specific for a given gene, are immobilised on a solid surface (for example a microscope slide), arranged in a known order, and are hybridised with a solution of labelled DNA or RNA molecules. The DNA or RNA molecules under investigation bind to complementary base pairs on the slide, and permit us to measure the amount of labelled DNA or RNA hybridised to each gene sequence. Sophisticated software is used to analyse the large amount of data generated. The ultimate aim is to group genes with similar expression patterns across all samples, and then group samples accordingly. Conventional biological or biochemical techniques are required to validate the data obtained with microarrays, and to verify whether the observed associations among genes are biologically relevant. PMID- 12741170 TI - [Ethnic and religious diversity of first year medical students at the Erasmus University Medical Center, Rotterdam]. AB - OBJECTIVE: To obtain information on the ethnic, religious and socio-cultural diversity of first-year medical students at the University Medical Center Rotterdam, the Netherlands. DESIGN: Anonymous questionnaire. METHOD: In December 2001, first-year medical students were given a questionnaire during a compulsory practical class. In the processing of the result, use was made of Statistics Netherlands' classification of allochthonous persons. Data were also collected on the student's mother tongue, self-reported command of Dutch and other languages, religious denomination and the education of their parents. RESULTS: The response rate was 90% (277/308); 175 women and 102 men. Seventy-four percent of respondents were rated as 'autochthonous' (both parents born in the Netherlands), 8% as 'western allochthonous' (at least one parent born in another western country) and 18% as 'non-western allochthonous' (at least one parent born in a non-western country). 'Allochthonous' students originated from 30 different countries, most frequently Surinam (n = 15), and spoke 26 different languages. Of the 'western allochthonous' students 70% considered themselves to be 'ethnic Dutch', of the 'non-western allochthonous' 65% considered themselves to be both Dutch and 'allochthonous'. Fifteen students did not speak Dutch with their parents and rated their command of Dutch to be significantly lower (score 8.9; scale 0-10) than that of the others. Forty-eight percent of the respondents did not consider themselves to be a member of any religious denomination, 18% were Protestant, 15% Roman Catholic and 7% Muslim. For 60% of the respondents, at least one parent had finished higher education. Parents of ethnic Turkish and Moroccan students had a significantly lower education than parents in all other groups. PMID- 12741168 TI - [Novel approaches; improved diagnostics and therapeutics with DNA microarrays. II. Applications]. AB - The most important (future) applications of DNA microarrays in medicine are based on three different molecular analyses: gene expression analysis, genotyping with a specific type of microarray (single nucleotide polymorphism chip (SNP chip)), and DNA sequencing to search for a well-known mutation in a certain gene. With gene expression analysis one can obtain a 'molecular signature' of a tissue by allowing tissue RNA to react with a DNA microarray. This information may help to refine disease classifications, to guide the choice of therapy and to find new therapeutic targets. Genotyping utilizes genomic DNA that, after digestion, reacts with a SNP chip to obtain an individual SNP pattern. An SNP is a change of a single base pair within a gene sequence that can sometimes influence the function of the gene product. SNPs are considered to be variations in the genome and should be distinguished from mutations. These variations can for instance provide information about the probability of a certain disease, or the effectiveness or side effect of a certain drug. Sequencing well characterized genes to search for mutations is used to screen for genetic abnormalities. This process can be accelerated with the aid of oligonucleotide arrays. With this tool an individual's risk of developing certain forms of cancer or monogenetic diseases can be estimated. PMID- 12741174 TI - [Summary of the Dutch College of General Practitioners' practice guideline 'Dizziness']. PMID- 12741173 TI - [The physician as an expert in medical negligence cases]. AB - Whenever a physician is charged with negligence, the professional opinion of another doctor is always required. The quality of the expert report is a decisive factor in the quality of the ruling by the adjudicator. Three main problems may arise here: the medical expert may push up the professional standard, experts may differ in opinion, and finally their judgment may be influenced by prior knowledge of the outcome (hindsight bias). To avoid these problems, a systematic approach is needed, taking into account the scientific context, the clinical context, the procedure followed, the degree to which the outcome corresponds with the predicted outcome, reappraisal of the facts and the organisational context. In particular, the problem of hindsight bias through prior knowledge of the (unhappy) outcome highlights the need for greater attention to scientific and procedural aspects in the formation of an expert opinion. PMID- 12741171 TI - [Severe diarrhea and eosinophilic colitis attributed to pinworms (Enterobius vermicularis)]. AB - In a 32-year-old woman suffering from severe diarrhoea, eosinophilic infiltration of colonic mucosa and a peripheral eosinophilia, microbiological investigations only revealed large numbers of Enterobius vermicularis (pinworm) in the faeces. Treatment with mebendazole resulted in a rapid resolution of symptoms and disappearance of the eosinophilia, which strongly suggested a causative role of this pinworm in the clinical syndrome of the patient. E. vermicularis is generally regarded as an innocent nematode, which at most causes perianal pruritus due to migration of worms from the colon and expulsion of eggs onto the perianal skin. Although the pinworm maturates and lives in the gut, gastrointestinal symptoms have seldom been reported. E. vermicularis infection should be considered in patients with unexplained eosinophilic enteritis. PMID- 12741172 TI - [Medical education in the Netherlands: little attention paid to the cultural diversity of patients]. AB - OBJECTIVE: To ascertain how much attention is given to cultural diversity in the eight medical faculties in Dutch universities. DESIGN: Interviews. METHOD: In the period January-June 2001 interviews were held with 76 people in medical faculties: policy makers, teachers and students. RESULTS: Medical courses scarcely devoted any attention to the fact that physicians need to deal with a multicultural society. Only the medical courses at the Free University of Amsterdam and Nijmegen University had included a compulsory module in this subject. The other universities were aiming for a more integrated treatment of the subject. However, for the time being, the acquisition of information about cultural diversity (specific diseases in different ethnic groups, different conceptions about health and disease and possibilities for solving communication problems) depended mainly on the personal interest of the teacher and the choices of the medical students. CONCLUSION: Cultural diversity receives little attention in the medical courses at the eight Dutch university medical faculties. A national approach to this problem is therefore recommended, with the setting up of an interfaculty workgroup to develop good teaching material on subjects related to cultural diversity. PMID- 12741176 TI - [Nuchal rigidity in children: meningitis or not?]. PMID- 12741175 TI - [Nuchal rigidity in children: meningitis or not?]. PMID- 12741177 TI - [The agrochemical arsenal versus the enemies of plants. General considerations]. AB - Plants are attacked, not only by various microorganisms, but also by other enemies, such as molluscs, nematods, mites, and insects. They have evolved complex and efficient mechanisms to defend themselves against pathogens (hypersensitive response, systemic acquired resistance) and herbivores (release of volatile compounds that attract predators of the herbivores, accumulation of proteinase inhibitors). Yet, the confrontation of the plants with their invaders can also turn to the advantage of the latter. In the past, the attacks of crops regularly brought about dramatic economic losses. From the World War II onwards, the development of organic chemistry associated with a growing awareness of the problems of agriculture has resulted in the production of a constantly growing number of plant protection products. They are currently divided into about ten classes, the herbicides, fungicides, and insecticides-acaricides making up more than 90% of the world market. Most of the agrochemical products put on the market over these last three decades are used in relatively low doses and have a more favourable toxicological and ecotoxicological profile than those of the former pesticides, many of which are now withdrawn from the market. Several more or less recent families are derivatives of metabolites from various organisms. Thus, the improvement achieved in the protection of crops is outstanding. However, one on the main side-effect is an environmental imbalance that has entailed a dependency on agrochemicals. Quite judiciously, alternative strategies (elicitors, genetic engineering, etc.) have been initiated or developed over the last decade. PMID- 12741178 TI - [Modes of action of agrochemicals against plant pathogenic organisms]. AB - The chemical control of plant pathogens concerns mainly fungal diseases of crops. Most of the available fungicides act directly on essential fungal functions such as respiration, sterol biosynthesis or cell division. Consequently, these compounds can exhibit undesirable toxicological and environmental effects and sometimes select fungal resistant strains. Plant activators are expected to provide sustainable disease management in several crops because the development of resistance is not expected. Considering the future, the discovery of novel antifungal molecules will reap advantage from throughput screening methodologies and functional genomics. PMID- 12741180 TI - Agronomic approach: cropping systems and plant diseases. AB - This paper deals with the effects of agronomic practices on parasite life cycles, and the design of integrated crop protection strategies. Cropping systems have a large effect on the size of the primary inoculum and its localisation, on the development and spread of epidemics, and on the coordination of the life cycle of cultivated plants and that of their parasites. They can disrupt ecological equilibria, either favouring or disfavouring the pathogens. By combining information concerning the effects of agricultural techniques on diseases and the physiological effects of diseases on growth and crop production, it is now possible to develop new crop management systems, in which the use of non-chemical methods for preventing diseases is a priority. However, the current knowledge need to be completed by studies on other scales, particularly of the effect of cropping systems on the genetics of disease populations integrating more completely the 'long-term' dimension of sustainable agriculture. PMID- 12741179 TI - [Contribution of cell and molecular biology and genetics to plant protection]. AB - Plants resist to the majority of their potential aggressors by opposing physical and chemical barriers: cell walls, secondary metabolites.... Phenomena of specific recognition between a plant variety and a pathovar induce on the one hand, a local (hypersensitive) reaction that tends to limit pathogen growth and, on the other hand, a cascade of signals that allows the activation of a non specific general (systemic) resistance. The contribution of genetics to the fight against pathogens depends on the natural variability that comes from the co evolution between plants and their aggressors. Many plant varieties resistant to one or several pathogens have been obtained and are cultivated. The use of biotechnology will facilitate the rapid generation of new, resistant cultivars and cultivars with multiple resistances. New methods in order to increase the efficiency and the durability of resistance are envisaged. PMID- 12741181 TI - [Implementation of new plant-health strategies]. PMID- 12741183 TI - Networks as constrained thermodynamic systems. AB - We show how a network of interconnections between nodes can be constructed to have a specified distribution of nodal degrees. This is achieved by treating the network as a thermodynamic system subject to constraints and then rewiring the system to maintain the constraints while increasing the entropy. The general construction is given and illustrated by the simple example of an exponential network. By considering the constraints as a cost function analogous to an internal energy, we obtain a characterisation of the correspondence between the intensive and extensive variables of the network. Applied to networks in living organisms, this approach may lead to macroscopic variables useful in characterising living systems. PMID- 12741184 TI - [Adaptations of the hoatzin (Opisthocomus hoazin) to leaf-eating. Morphological characteristics and functional features of its bill and hyoid apparatus]. AB - The hoatzin remains one of the most enigmatic birds. A morphofunctional analysis of its bill and hyoid apparatus throws new light on its feeding adaptation as well as on its systematic relationships. Bony and muscular skull, rhamphotheca, palate, and hyoid apparatus were described in details. Though keeping into the general organisation pattern found among Neognathae (except Galliformes), bill and hyoid apparatus of the hoatzin displays a series of species-specific features, some unique among birds. This species appears particularly well adapted to tear of leaves and process them inside the bill before ingestion. Because of very important anatomical and thence functional differences in bill and hyoid structure, any close relationship between the hoatzin and Galliformes cannot be envisioned. Such a hypothesis would implicate a counter-selective evolutionary reversion. PMID- 12741185 TI - A chemotaxonomic method to quantify phytoplankton groups in freshwater lentic mesocosms: an approach including chlorophyll a breakdown products. AB - The use of HPLC methods to determine and quantify phytoplankton population composition, is sometimes less time-consuming than microscopic identification. However, its general application poses problems since high discrepancies between chlorophyll a calculated using chemotaxonomic methods and direct measurements were noticed. For instance, chemotaxonomic protocols generally employed can lead to a poor estimation of total and relative abundance when high amounts of chlorophyll a breakdown products are present. Therefore, we propose a new approach to calculate relative abundance of algal groups in a phytoplankton population, based on integration of these degradation products in the chemotaxonomic assessment in lentic and shallow freshwater ecosystems. PMID- 12741182 TI - [Homologous recombination and gene targeting]. AB - Gene therapy and the production of mutated cell lines or model animals both require the development of efficient, controlled gene-targeting strategies. Classical approaches are based on the ability of cells to use homologous recombination to integrate exogenous DNA into their own genome. The low frequency of homologous recombination in mammalian cells leads to inefficient targeting. Here, we review the limiting steps of classical approaches and the new strategies developed to improve the efficiency of homologous recombination in gene-targeting experiments. PMID- 12741186 TI - [A new approach to biodiversity: morphological plasticity of a fresh-water diatom]. AB - Population responses of a planktonic freshwater diatom, Asterionella formosa Hassal, to the hydrodynamic anthropic disturbances were studied at the landscape scale, along a series of nine reservoirs, for a period of 18 months. The analysis of biotic descriptors as cell abundance, cell length and architecture has shown a strong morphological plasticity of this diatom. The morphological variability of A. formosa in response to hydrodynamic conditions favours the colonization of lacustrine freshwater ecosystem for this species. The determination of architectural indices allowed us to distinguish a modification of biodiversity along a disturbance gradient for population level. PMID- 12741187 TI - Dispersal and fighting in male pollinating fig wasps. AB - For more than two decades, it has been the dogma that the males of pollinating fig wasps do not fight and that they only mate in their native fig. Their extreme degree of local mating leads to highly female biased sex ratios that should eliminate the benefits of fighting and dispersal by males. Furthermore, males sharing a fig are often brothers, and fighting may be barred by kin selection. Therefore, theory supported the presumed absence of fighting and dispersal in pollinating fig wasp males. However, we report here that in pollinating fig wasps, fighting between brothers evolved at least four and possibly six time, and dispersal by males at least twice. This finding supports the idea that competition between relatives can cancel the ameliorating effects of relatedness. The explanation to this evolutionary puzzle, as well as the consequences of male dispersal and fighting, opens the doors to exciting new research. PMID- 12741188 TI - [Total nephrectomy in children: 11 years of experience in 80 cases]. AB - Nephrectomy in childhood is an ultimate indication. It is realised in case of severe deterioration of the renal function or neoplastic affection of the kidney. We report a retrospective study on a series of 80 patients. Mean age was 5 years with extremes from 2 months to 14 years. Abdominal mass, pain, infection and haematuria were the mostly encountered revealing symptoms. The main aetiology was nephroblastoma. No laparoscopic nephrectomy was realised. Follow-up was uneventful except for the troubles existing prior to surgery. PMID- 12741189 TI - Effects of intraperitoneally administered vitamin E and selenium on calcium oxalate renal stone formation: experimental study in rat. AB - Forty-eight Wistar rats were treated for 3 weeks with water containing 0.7% ethylene-glycol and divided into four groups. The first group, used as control, has received sodium chloride at 1 ml/100 g BW daily. The second group was intraperitoneally injected with selenium at 10 micrograms/d per 100 g BW as NaSeO3 for 3 weeks. The third group was intraperitoneally administered with 15 mg Vit E/d per 100 g BW as alpha-tocopherol acetate for 3 weeks. The last group was simultaneously administered vitamin E and Se at the same doses and periods as the precedent groups. One day before the end of the treatment, each animal was placed in a metabolic cage for collection of 24 h urine samples and determination of urinary creatinin, urea, calcium, magnesium, phosphate and oxalate levels. Immediately thereafter, all the rats were anesthetized and aortic blood was collected to determine the same parameters as in urine. The kidneys were also removed to determine calcium oxalate deposits, dry weight and to conduct a histological examination. Our results showed decreased ionic product and increased magnesium fractional reabsorption in the group receiving only selenium and in the group receiving selenium in combination with vitamin E, in comparison with the control animals. In view of the knowledge concerning the same protective action of Vit E and selenium, regardless of tubular membrane alteration, the absence of any inhibitory effect of Vit E on calcium oxalate formation suggests that selenium, like other minerals, could be stuck onto the crystal surface and would inhibit induction of new crystals, growth and aggregation. PMID- 12741190 TI - [Emphysematous pyelonephritis: a case report]. AB - Diabetic patients with urinary tract infections had a certain risk of developping emphysematous pyelonephritis with gas producting bacteria. This disease was accompagned with high mortalite. We will report one case who was treated successfully by high dose antibiotic regim and emergency nephrectomy. PMID- 12741193 TI - [Giant hydronephrosis: two case reports]. AB - Giant hydronephrosis is uncommon. Ureteropelvic obstruction represent the more frequent cause. In 1939, Stirling defined it as the presence of more than 1000 ml of fluid in the collecting system. The Authors report two cases with ureteropelvic obstruction causing giant hydronephrosis. The treatment includes pyeloplasty in the first case and nephrectomy in the second case. PMID- 12741191 TI - [Emphysematous pyelonephritis: review of the literature concerning a case report]. AB - Emphysematous pyelonephritis is defined as the presence of gas-producing bacteria in the kidney and in peri-nephretic areas. Even if it is rare, the mortality rate of this affection is between 50% and 90%. The E. coli is responsible in 60% of the cases. We report a case of a 50 years old male patient, with under diagnosed diabetics, how is admitted with pains in the left flank, fever, troubled urine, hematuria and worsening of general state. The diagnostic of emphysematous pyelonephritis was confirmed by CT Scan. In spite of adapted antibiotherapy to the renal function, insulinotherapy and correction of hydro-electrolytic troubles, the patient died with septic shock associated to digestive bleeding. Based on this case and a review of the literature, the authors describe the different features of this disease. Only an urgent nephrectomy after a short reanimation can improve the prognostic. PMID- 12741194 TI - [Macronodular adrenal hyperplasia: a case report]. AB - This case report describes a 53-year old man with Cushing's syndrome due to Adrenocorticotropic Hormone (ACTH) independent bilateral adrenocortical macronodular hyperplasia (AIMAH). His plasma cortisol showed no diurnal rhythm and was unsuppressable by high-dose dexamethasone. Plasma ACTH was undetectable and did not respond to corticotrophin-releasing hormone. The aetiology of this rare disease remains uncertain. The treatment is based on bilateral adrenalectomy. PMID- 12741192 TI - [Kidney cancer: report of 47 cases]. AB - The authors report a series of 47 cases observed over a 15-years period in the department of carcinological at the national oncological institute of Rabat. Patients consisted of 16 mean and 21 women (57%), with a mean age of 49 years old (range 29 to 70 years). The clinical features were polymorph, firstly loin pain (81%), hematuria is the next (25%). A lumbar mass (10%). The diagnosis was established by ultra-sonography and CT-scan in all the patients. The mean tumour diameter was 10 cm (5-17 cm), it were located in down pole in 53%. The surgical procedure consisted of radical nephrectomy and regional or hailer lymph node dissection in 46 patients. The post operative course was marked by one death due to pulmonary metastases. The mean follow-up was (2 to 15 years). Asynchronies metastases occurred in 6 patients after a mean interval (9 to 36 months). The overall 5 year survival was 100% PT1, 78% PT2, 34% PT3. PMID- 12741198 TI - [Bladder stone surrounding a foreign body: a case report]. AB - The bladder can be the site of various foreign bodies. We report one case of bladder stone including a foreign body in a 24 years old man with a psychomotor deficiency who was admitted for pyuria, block miction and bladder symptoms. The pelvic X-Ray film showed a bladder stone including a sewing needle. We analysed the diagnosis, aspect and therapeutic management of this case. PMID- 12741197 TI - [Small cell neuroendocrine carcinoma of the bladder. A new case report]. AB - This rare but aggressive bladder tumour presents the morphological and immunohistochemical characteristic common to all neuro-endocrine tumours observed in other organs. The authors report a case of primarymalignant neuro-endocrine bladder tumour. Combined surgery-cisplatin-based adjuvant chematherapy is recommended. PMID- 12741199 TI - [Metastatic cerebral abscess from Klebsiella pneumoniae after extracorporal shock wave lithotripsy for kidney stone (a case report)]. AB - Extra-corporeal Shock Wave Lithotripsy (ESWL) is widely used to treat urinary calculi. With increasing numbers of ESWL, more patients prone to infectious complications. A 62-year-old man presented with cerebral abscess following ESWL. The authors discuss presenting conditions, risk factors and preventive measures. PMID- 12741196 TI - [A rare cause of urinary obstruction: urogenital tuberculosis]. AB - The authors reported a rare cause of urogenital tuberculosis complicated by an obstructive acute renal failure in 44 years old man with solitary anatomic kidney. The authors insisted of using the upper urinary tract opacification by percutaneous nephrostomy for diagnosis, the urogenital tuberculosis with this exploration, we can suspected the tuberculosis by abnormalities of the radiologic imagine, and confirmed the koch bacilli urinary into urinary tract. The upper chance of positives of finding koch bacilli in higher than urinary bladder. PMID- 12741195 TI - [Retroperitoneal fibrosis with pelvic extension: a case report]. AB - The retroperitoneal fibrosis is a rare pathology, whose physiopathologic mechanism is poorly known. It characterizes by the training of a benign fibrous prevertebral plate. This plate is in rule limited between the renal pedicle in high and the promontory anointed down. However, extensions to the great peritoneal cavity, the mediastinum and especially the pelvis have been brought. Authors bring a case of retroperitoneal fibrosis at a woman of 54 years, that spreads to the pelvic region complicated of an obstructive renal insufficiency. The ureteral catheterism has allowed the standardization of the renal function. The uroCT-scan has allowed to make a complete lesion statement. The patient has been operated on with realization of ureterolysis and ureteral intraperitonization. Through this observation, therapeutic and diagnostic aspects will be discussed. PMID- 12741204 TI - Adolescent leisure across European nations. PMID- 12741200 TI - Korean adolescents' "examination hell" and their use of free time. AB - A 15-year-old girl's daily life is a vivid testimony to the hardships the Korean high school student is faced with. She leaves home for school at 7 A. M., taking two lunch bags, one for supper, with her. After regular classes end at 5 P.M., she attends the "autonomous study classes" studying by herself until 10 P.M. The exhausted young girl returns home at about 10:30 P.M. and gets to bed around midnight at the earliest. Under the current system, this will be her life for three years. Of course, there is no guarantee she will be able to enter the university she wants. She has no family life, except for Sundays, and they hardly ever see each other, let alone get together at the dining table for dinner. People call this "ipsi chiok," entrance examination hell. ["High School Students Deprived of Spring," 1996, p. 13]. PMID- 12741202 TI - Cultural continuity amid social change: adolescents' use of free time in India. PMID- 12741201 TI - Japanese adolescents' free time: juku, bukatsu, and government efforts to create more meaningful leisure. PMID- 12741205 TI - Italian adolescents and leisure: the role of engagement and optimal experience. PMID- 12741203 TI - Adolescents' leisure time in the United States: partying, sports, and the American experiment. PMID- 12741209 TI - Proof positive. PMID- 12741206 TI - Contextual constraints on adolescents' leisure. PMID- 12741208 TI - Tissue from stem cells may help repair joints. PMID- 12741207 TI - Access to health care elusive for Americans with disabilities. PMID- 12741213 TI - The fall factor. PMID- 12741210 TI - Shopping for an ultrasound machine. PMID- 12741211 TI - On the road again. PMID- 12741214 TI - The McKenzie approach. PMID- 12741212 TI - Insightful options. PMID- 12741215 TI - Rehab's medicine cabinet. PMID- 12741217 TI - Appearances can be deceiving. PMID- 12741216 TI - Maximizing energy efficiency. PMID- 12741220 TI - FCEs: to use or not to use? PMID- 12741219 TI - Reeve's therapy leads to partial recovery. PMID- 12741218 TI - Are you provider-based? PMID- 12741221 TI - Making moving easier. PMID- 12741222 TI - A balanced approach. PMID- 12741223 TI - 10 tips for safe mobility in the bariatric population. PMID- 12741230 TI - The golden privacy rule. PMID- 12741225 TI - Opportunity knocks. PMID- 12741224 TI - The silent disease. PMID- 12741226 TI - Delving into design. PMID- 12741229 TI - Do you yearn for private practice? How to set up your own shop. PMID- 12741231 TI - Light at the end of the tunnel. PMID- 12741234 TI - To buy or not to buy. PMID- 12741233 TI - Walking off the pain. PMID- 12741236 TI - A joint dilemma. PMID- 12741235 TI - No room for discomfort. PMID- 12741227 TI - The prevention struggle. PMID- 12741232 TI - Meeting of the minds. PMID- 12741228 TI - Play time. PMID- 12741241 TI - CORF ideas. Comprehensive outpatient rehab facilities are becoming profitable investment options. PMID- 12741237 TI - The CPM challenge. PMID- 12741242 TI - Women first. PMID- 12741243 TI - Orthotic options. PMID- 12741238 TI - Pointing to prevention. PMID- 12741240 TI - Health care and the 108th Congress. PMID- 12741244 TI - Finding the comfort zone. PMID- 12741247 TI - The FNS project. PMID- 12741245 TI - The right fit. PMID- 12741239 TI - Compliance 101. . How effective is your company's HIPAA compliance program? PMID- 12741246 TI - A child's first seating system. PMID- 12741248 TI - A changing environment. PMID- 12741251 TI - Scheduling difficulties. PMID- 12741249 TI - HIPAA Smarts. Top 10 privacy musts. PMID- 12741253 TI - On the right track. PMID- 12741252 TI - Out to lunch on the $1,500 cap. PMID- 12741250 TI - Community creation. PMID- 12741255 TI - A test of nerves. PMID- 12741257 TI - Strength over surgery. PMID- 12741256 TI - Losing friction. PMID- 12741258 TI - Let the games begin. Bush proposal opens Medicare debate in the 108th Congress. PMID- 12741261 TI - A sea change for Spaulding. PMID- 12741259 TI - Confusing CORF concepts. PMID- 12741254 TI - Rules of the road. PMID- 12741260 TI - PT vs OT. PMID- 12741263 TI - Stand up and function! Wheelchair-dependent clients benefit both medically and functionally from stander use. PMID- 12741262 TI - Taking a stand. Evaluation, justification, and documentation for standers. PMID- 12741267 TI - Rolling, rolling, rolling. PMID- 12741269 TI - Gait analysis and Parkinson's disease. PMID- 12741273 TI - Chest physical therapy. PMID- 12741264 TI - Older does not equal obsolete. PMID- 12741265 TI - The skinny on splinting. PMID- 12741266 TI - Rehab lands a starring role. PMID- 12741272 TI - Chest physical therapy (CPT). PMID- 12741270 TI - Relationship construction. Networking is a boon to practice building. PMID- 12741274 TI - Innovative intervention. OT students help homeless people recover the skills they need to return to mainstream society. PMID- 12741268 TI - Breaking with tradition. PMID- 12741275 TI - Wading through the possibilities. PMID- 12741271 TI - The ABCs of PPS and LTACHs. PMID- 12741277 TI - The positive spin. PMID- 12741278 TI - Covering your bases. PMID- 12741276 TI - Getting back to the daily grind. PMID- 12741279 TI - Craig gets mobile. PMID- 12741280 TI - Beyond basic ADLs. PMID- 12741284 TI - Protecting patient privacy. PMID- 12741285 TI - When patients don't show up. PMID- 12741282 TI - Wounds and warmth. PMID- 12741287 TI - [Comparative study of respiratory mechanisms between professional lyrical singers and beginners]. AB - The apprenticeship for classical singing requires years of training due to its complexity. This training is directed at the different levels of the vocal apparatus: the source of energy or air flow (lungs), the vibrator (vocal cords) and the resonators (bucco-pharyngeal cavity). For this study we have concentrated on the first level, that is, respiration. When a greater demand of vocal activity is required respiration becomes more sustained and more complex; this is when difficulties may be observed. Singers overcome these difficulties through different strategies according to their level of vocal training. Our study compared respiratory strategies used by professional singers and singing students (first year conservatoire students) during singing tasks; none of the subjects had any history of vocal pathology. Electromyographic analysis of muscle activity was registered for the rectus abdominals muscle, external oblique muscle and transverse muscle with surface electrodes. Kinetic analysis was used to measure thoracic cage displacement: pressure sensitive belts were placed around the pubis, the epigastric region and thorax. The subjects were asked to perform two vocal tasks: vocalization of a sustained "i" and singing of an extract of a vaccai. Results were compared between the 7 professional singers and the 6 singing students. They allowed us to confirm the existence of distinct respiratory strategies according to the level of vocal training, as described in literature. Professional singers inhibited the activity of their rectus abdominals muscles during singing and used mostly their external oblique and transverse muscles, thus, sustaining an expanded ribcage and a longer expiratory breath. Singing students, on the other hand, worked intensively all the abdominal muscles leading to a collapse of the ribcage. Management of air was more difficult and breath was shorter. Future studies will consider a larger population sample in order to define quantitative parameters that might allow a significant differentiation of respiratory strategies between professional singers and singing students. PMID- 12741283 TI - Ergo MOMics. PMID- 12741286 TI - [Non-linear model of glottic vibration. Potential clinical implications]. AB - Glottic vibration is governed by a certain number of physical laws which in general correspond, as far as myo-elastic theory is concerned, with the laws of passive vibration of an elastic body under the influence of the expired air. Some of these laws are non-linear, especially those that deal with the exchange of energy between the expired air and the vocal folds. This non-linearity is specially important when it comes to the key concepts of the phonatory threshold and the passive synchronisation of the vocal cords. Based on their experimental work, which has been presented previously, the authors propose a mixed model, which is both linear and non-linear, like the oscillators of Van der Pool. This model is known as Slip-Stick. PMID- 12741289 TI - [Voice breaking phenomenon in female adolescents]. AB - The title at the beginning of this study, "voice-breaking in adolescent females" has given rise to some lively and forceful reactions. It is a provocative concept for those who contest its validity. For others, this is quite a natural phenomenon to study: "it is widely recognised that the breaking of the voice forms part of the secondary sex changes, and that as the larynx enlarges, the voice of the young girl necessarily changes with it". PMID- 12741290 TI - [Importance of tussometry in unilateral laryngeal paralysis]. AB - Cough is a physiological gesture witch requires a perfect laryngeal competence. Tussometry was described as a reliable and reproducible method of analysis based on the measure of oral air flow rate during cough. This test allows to quantify the efficiency of laryngeal occlusion in normal situation and in case of unilateral laryngeal paralysis. The measure of the time lag between the beginning of the cough and the maximal air flow rate (peak value time) is the most reliable parameter (8). Present study is designed to asses the reproducibility of tussometry realized on a vocal analysis workstation EVA II and to test the efficiency of intracordal injection of autologus fat in case of unilateral laryngeal paralysis following thoracotomy. 10 control subjects were recorded at two moments. 6 patients presenting unilateral laryngeal paralysis following thoracic surgery were recorded before and after injection of autologus fat in the paralysed vocal cord. Reproducibility of tussometry is good (r = 0.96). The mean peak value time is significantly improved (p = 0.048). In conclusion, tussometry is a reliable routine test on EVA III workstation. Intracordal autologus fat injection improves occlusive function of larynx during cough in case of unilateral paralysis. PMID- 12741291 TI - [Intelligibility of French consonants after partial supra-cricoid laryngectomy]. AB - INTRODUCTION: The purpose of this study was to determine what patterns of perceptual confusions characterize the voice of patients after supracricoid partial laryngectomy (SCPL) by the identification tests of French consonants. We evaluated the voicing distinction. METHODS: Ten male patients were recorded 18 months after SCPL. Audio recordings of the 16 French consonants in a syllabic context (CV) produced by each talker with three repetitions were presented to three expert listeners. The listeners transcribed their responses using an open response paradigm. Listeners' pooled responses were converted to confusion matrices. RESULTS: Voicing features were altered with a shift of voiced consonants towards voiceless consonants (24%), predominantly for stop consonants. CONCLUSION: Consonant articulation appears to impose certain constraints on voicing ability of SCPL patients, since voiced consonants are predominantly perceived as voiceless consonants. Presumably, this poor voicing ability is the direct consequence of the mechanical properties of the neoglottis that are far different from those of the vocal folds. Moreover, assessing consonant identification and intelligibility should help to improve voice therapy and efficiency of speech. PMID- 12741288 TI - [Attention deficits in the school aged stuttering child: constituent trait or comorbidity]. AB - A first account of a prospective study of attention deficit disorders among school-age stutterers--in the settings of a fluency clinic newly opened in a large general hospice--has led to the evidence of a significant association between impulsivity and stuttering. It seems therefore necessary to include attention tests among the subtests of our assessment protocols. Ways of helping the young stutterers with ADD are later thoroughly described. PMID- 12741281 TI - The tribulations of transferring. PMID- 12741297 TI - [Echo reading and stuttering]. AB - Echo-reading is a practical therapeutic tool which has been proposed for use in stuttering, most especially when the functions of rhythm and association are disrupted in stuttered speech. After a period of preparation in which the mechanical work involved in speech is integrated, it is proposed that there should follow a period of listening and association, which serves to re-establish the spoken word with its meaning and expressivity. The subject undergoes rehabilitation in a living language which is correlated with the imagination, and with that Other quality which always forms a part of speech. PMID- 12741294 TI - [Different therapeutic suggestions used in 4 types of laryngopathies presenting in professional singers]. AB - Four clinical cases involving non-surgical approaches to disorders usually treated surgically are considered. The emphasis is placed on the collaboration of artists with laryngologists combining their musical and medical training in their daily work at their office and an alternative clinical management of disorders. PMID- 12741292 TI - Phonatory threshold pressure in a healthy population before and after aerosol treatment, a preliminary study. AB - The viscosity of the surface mucus of the vocal cords is one of the important elements for good laryngeal functioning. It has been demonstrated that inhalation of hydrated air increases the phonatory threshold pressure by decreasing viscosity of the mucus (1) leading to a more regular vibration that can be appreciated by jitter (2). In an attempt to correlate the concepts of tissue viscosity and surface mucus considering the theoretical model of vibration we measured the phonatory threshold pressure in 6 healthy female subjects before and after aerosol treatment. We were able to demonstrate that the pressure threshold is lower (3.15 hPa) after aerosol treatment than before (3.79 hPa) and this was statistically significant (p: 0.041). The discussion is based on this decrease of mucus viscosity applied to the physiological concepts necessary to understand glottic vibration. PMID- 12741301 TI - [The prophylactic defibrillator after the MADIT II study. The glimpse of a turning point?]. PMID- 12741298 TI - [The patient, his guides, his therapists. Role of the orthophonist]. AB - It is the purpose of this article to elucidate certain aspects of the speech therapy management of the patient who is a professional voice user, as he progresses towards a better use of his voice. Based on personal case experience, the author seeks to alert the reader to the role of supportive listening, empathy and personal qualities of the therapist which enable him/her to come to a better understanding of the patient's problem. PMID- 12741296 TI - [Is stuttering a functional dystonia?]. AB - For some years the dystonias have been the subject of major studies and, as far as the generalised dystonias are concerned, of major therapeutic advances. The opposite is true of the so-called focal or functional dystonias, which include conditions such as Meige's syndrome, spasmodic torticollis, writer's cramp, dystonias using instruments especially in musicians, and spasmodic dysphonia. For the last group, the term functional dysphonia would seems to us to be more appropriate. It would appear that what is involved is a disorder not of a muscle group, but rather of a function. Consequently stuttering can, in our opinion, be legitimately considered as a dystonia affecting the articulation of speech, within the global context of a new neurological grouping which could be called 'dysfunctional neurology'. PMID- 12741299 TI - [The phoniatrist-orthophonist relationship: a team of 2 or a team of 3?]. PMID- 12741295 TI - [Basic principles for managing and post-operative follow-up in phono-surgery]. AB - The management of the voice in patients with benign lesions of the larynx causing chronic dysphonia follows certain simple rules, on which depend the final functional result. It also requires the closest collaboration between the voice surgeon and the speech therapy team. The indication for surgery rests on the most precise diagnosis of the condition, after examination of the larynx with the rigid endoscope and video-stroboscopy of the cord movements. When a final decision for surgery has been taken, it is essential that the patient should be thoroughly prepared for surgery if the post-operative phase is to pass smoothly, which is the criterion for a good functional recovery. The surgery must be both precise and thorough, with due respect to the structural elements of the vocal folds, and must involve the minimum of resection. Afterwards, a period of complete voice rest is mandatory, before the voice rehabilitation can commence. Finally, post-operative follow-up will be the best guide for selecting the appropriate speech therapy, which will allow the patient to overcome the anatomoical and functional changes brought about by surgery, to recover the potential qualities of his voice, and to resume his vocal and professional activity under the best conditions. PMID- 12741300 TI - [Pedagogy of inspiration]. AB - We often hear that the wisest about vocal reeducation and more specifically about the problem of phonatory breathing is to focus only on the exhale, leaving the inhale automatic and involontary. This carefull approach is explained by the fact that the air-intake is a normally automatic and inconscious gesture in the speech which risks indeed to become unnatural if voluntarily controlled. A training that would preserve the natural nature of the gesture thanks to the practice of relaxation, permits however to avoid this difficulty and to lead to a better rehabilitation of the phonatory behaviour. PMID- 12741293 TI - [Anti-viral injectable treatment (cidofovir) in laryngeal papillomatosis]. AB - Laryngeal papillomatosis, due to type 6 and 11 papova-virus A, causes devastating lesions leading to difficult clinical situations (severe dysphonia, or laryngeal dyspnea). Recurrence requires repeated endoscopy with CO2 laser treatment to keep the airways free and prevent the lesions spreading. In patients presenting aggressive papillomatosis, such repeated intervention causes irreversible lesions which have a very negative impact on the vocal prognosis. In this context, developing a form of minimally invasive surgery would help avoid vocal sequelae as far as possible. Thus, anti-viral agents can be injected directly into the lesion per-operatively so as best to preserve the healthy mucosa and muscles, thereby managing the lesions with precision and less iatrogenic impact than with CO2 laser. Twenty six patients since 1998 have undergone Cidofovir endoscopy. Total remission was achieved in eight of them (31%), after between two and eight interventions. Twenty seven (65%) showed clinically significant partial remission. Such positive results were obtained in both adults and children. Associated lesion excision was required in cases of obstructive or persistent papilloma. Intralesion Cidofovir injection thus seems to have proved highly effective in the clinical management of laryngeal papillomatosis. Combined Cidovir injection and surgical excision remains necessary in case of large or persistent papillomas. These results have lead us to indicate this procedure as primary treatment for laryngeal papillomatosis in adults and children. PMID- 12741302 TI - [Prognostic value of thallium 201 myocardial scintigraphy with dipyridamole before peripheral arterial surgery]. AB - The evaluation of patients who are candidates for peripheral arterial surgery is difficult. The aim of this study was to show that dipyridamole stress scintigraphy could be a prognostic aid for patient selection. Between 1991 and 2000, 275 patients underwent dipyridamole stress myocardial scintigraphy before peripheral arterial surgery of the lower limbs (49%), the aortic (33%) or carotid arteries (18%). A perfusion defect was observed in 145 patients suggesting myocardial ischaemia in 79 cases and myocardial infarction in 66 cases. Twenty seven of the 79 ischaemic patients underwent a preoperative coronary revascularisation. The operative adverse coronary events (5%) were: 7 non-fatal myocardial infarctions and 7 acute coronary syndromes. The 79 ischaemic patients had a higher risk of adverse coronary events: 11% (ischaemia) versus 3% (no ischaemia) (p < 0.01). Myocardial scintigraphy allowed stratification of patients with an intermediate risk of Eagle's score into high coronary risk (15%, ischaemia) or low coronary risk (2%, no ischaemia) (p < 0.01). The extent of the ischaemia was associated with a higher risk of adverse coronary events: 4 zones (20%) versus 1 zone (5%) (p = 0.02). Preoperative coronary revascularisation tended to reduce the risk of adverse coronary events from 15% to 4% (p = NS). Myocardial ischaemia (p < 0.0001) and left bundle branch block (p = 0.002) were the two predictive factors of an adverse operative coronary event. Thallium dipyridamole myocardial scintigraphy with a high negative predictive value (97%) is a useful tool for the identification of high risk patients for whom an aggressive preoperative therapeutic strategy may be beneficial. PMID- 12741303 TI - [Valvular replacement with the Bicarbon mechanical valve. Results after 7 years in 104 patients]. AB - OBJECTIVE: Study of the medium term results of aortic and mitral valve replacement with the Bicarbon' prosthesis. METHOD: From 1990 to 1996, 109 valves were implanted (70 in aortic position, 31 in mitral position and 4 double replacements). The average age was 61 years and 75% were male. According to the NYHA, 59% of patients were stage III or IV. The average pre-operative ejection fraction was 59.6%. There was re-intervention in 21.1% of patients and 35.3% had an associated procedure during the intervention. RESULTS: The average follow up was 5.4 +/- 1.98 years in 98 patients (that is 522 patient years). One patient died post-operatively and 19 died later. The overall survival at 7 years was 69.4 +/- 6.3%. Complications, expressed in patient years, were 1.15% for thrombo embolic complications, 2.1% for haemorrhagic complications. 0.38% for endocarditis, 1.72% for non-infectious peri-prosthetic leaks, and 0.76% for re interventions. At 7 years, the absence of thrombo-embolic, haemorrhagic, endocarditis, and re-intervention complications was 91.8 +/- 4.2%, 85.3 +/- 4.8%, 95.8 +/- 3.2%, 93.8 +/- 3.5% respectively. According to the NYHA, 95% of patients were in stage 1 or II (p < 0.001). CONCLUSION: Valvular replacement in the aortic or mitral position with the Bicarbon' valve is satisfactory as much in terms of survival as of clinical complications. PMID- 12741304 TI - [Coronary angioplasty using 5 or 6 F guide catheters. Results of a comparative, monocentric, prospective randomized study]. AB - Improvement of balloon and stent profile allows to use smaller-diameter catheters to avoid vascular complications by reduce the size of puncture site. First studies using 5 F guiding catheters showed good results in term of safety and feasibility. The authors performed a prospective, randomised study to define exactly the place of such small catheter compared to 6 F approach. One hundred forty eight patients were randomised, 77 in the 5 F group. The success rate per lesions was not significantly different but was less in the 5 F group (91.1 vs 96.5%). The crossover to 6 F allows good results in mainly cases. Limitations are unstable back up, worse opacification, and inability to use covered stent and to treat bifurcation lesions. There are no advantages in term of vascular complications. So, these catheters seem to be limited to treat simple lesion, allowing using the same sheath immediately after coronarography, but these results must be confirmed in a large, multicentric study. PMID- 12741305 TI - [Infectious endocarditis. A study of 50 patients in a non-university hospital]. AB - The authors report the results of a single centre study of 50 consecutive patients (average age 66 +/- 14 years; 36 men), admitted between 1992 and 2001 to a peripheral hospital for infectious endocarditis (IE). The median interval to diagnosis was 57 days. There was an underlying cardiac disease in 52% of cases, usually valvular (42%). The site of the IE was the mitral valve in 21 cases, the aortic valve in 19 cases, mitro-aortic valves in 5 cases, native tricuspid valves in 2 cases and pacing catheters in 4 cases (associated with valvular endocarditis in one patient). The causal organism was usually a streptococcus (60%, including 28% of streptococcus bovis), or a staphylococcus (22%): no organism could be found in 7 patients. The average follow-up was 33 +/- 30 months: surgery was indicated in half the patients and 3 patients were turned down because of their poor general condition. In all, 34% of patients died (24% of their IE) in a median interval of 6 months, mainly from infectious or haemodynamic complications. Poor prognostic factors were: age > 70 years, "blind" antibiotic therapy, large-sized vegetations, embolism and renal failure. These data, comparable to the results observed in large series in the literature, underline the importance of multi-disciplinary management of IE and strict prophylaxis. PMID- 12741306 TI - [The value of repeated determinations of brain natriuretic peptide for the diagnosis of unstable angina]. AB - The diagnosis of unstable angina (troponine undetectable) is often difficult in the absence of electrocardiographic changes after suggestive chest pains. The object of this study was to analyse the kinetics of Brain Natiuretic Peptide (BNP) during acute coronary syndromes (ACS) without ST elevation. Plasma BNP was measured every 6 hours for 48 hours in 65 patients admitted for suspicion of ACS without ST elevation and without clinical, radiological or echocardiographic signs of left ventricular dysfunction. The results of BNP measurements were masked until the final diagnosis was established on the usual investigations (ECG changes, troponine I values, myocardial scintigraphy, coronary angiography). These investigations identified 3 groups of patients: non-Q wave infarction (group A: 19 patients), unstable angina (group B: 21 patients) and non-coronary chest pain (group C: 25 patients). The peak BNP was significantly higher in groups A (210 +/- 172 pg/ml) and B (152 +/- 159 pg/ml) than in group C (16 +/- 14 pg/ml). However, the BNP was normal or only slightly increased (< 50 pg/ml) in 25% of cases of ACS. Analysis of the kinetics of BNP was much more discriminating: early increase after the pain, peak between the 14th and 24th hours (19th hour on average), followed by a progressive decrease. The kinetics were identical in Groups A and B, contrasting with the flat profile of the curve in group C. A change of > 20 pg/ml in BNP was a better criterion of ACS with a diagnostic accuracy > 90% than increased troponine (group A) or undetectable troponine (group B). The authors conclude that BNP kinetics is a new and reliable diagnostic marker of unstable angina when the usual criteria of ACS are not present (notably a normal ECG and undetectable troponine). PMID- 12741307 TI - [Treatment of left intraventricular thrombosis with low molecular weight heparin]. AB - Following the discovery of a left intra ventricular thrombus (LIVT), the classical approach consists of treatment with non-fractionated heparin (NFH) followed by oral anticoagulants. The use of NFH for this indication has only been evaluated in one open, non randomised study of 23 patients with no control group. Low molecular weight heparins (LMWH) have not been the object of any study although they are routinely used by certain teams. The objective of this study was to evaluate the feasibility of the use of LMWH in the treatment of left intra ventricular thrombus. This was an open, non randomised prospective study. All patients having a newly diagnosed LIVT between September 2000 and September 2002 were treated with enoxaparine (100 IU/kg twice daily) for an average duration of 13 days; replacement with fluindione was started on the fifth day. The progression of the LIVT was followed using twice weekly transthoracic echocardiography for 3 weeks. RESULTS: 19 LIVT were discovered in 2 years (13 complicating an anterior infarct and 6 with a dilated cardiomyopathy). The average area was between 2.64 +/- 0.41 cm2 and 0.43 +/- 0.21 cm2 (p < 0.0001). Thirteen out of 19 thrombi disappeared with treatment (68.5%). There was no thrombocytopenia or haemorrhage. One transient ischaemic attack was noted. CONCLUSION: This preliminary work shows that LMWH are well tolerated and effective to make a thrombus disappear or to reduce its size. PMID- 12741308 TI - [Intra-aortic balloon pumping and variations in flow in the dilated coronary artery in acute myocardial infarction]. AB - The object of this study of acute anterior myocardial infarction uncomplicated by cardiogenic shock, a context in which the role of intra-aortic balloon pumping (IABP) remains controversial, was to analyse the effects of IABP on coronary flow in the culprit artery. Twenty-one patients admitted for angioplasty in the acute phase of anterior myocardial infarction were included. The IABP was performed in 6 patients (Group 1) because of clinical signs of cardiac failure. Fifteen patients (Group 2) had no signs of cardiac failure. Coronary flow velocity was recorded by a Doppler catheter after successful angioplasty. The following parameters were analysed: average peak velocity (APV), average diastolic peak velocity (ADPV), average systolic peak velocity (ASPV), diastolic to systolic velocity ratio (DSVR) and maximum peak velocity (MPV). Intra-aortic balloon pumping was associated with an increase in the diastolic indices (APV 17.9 +/- 3.5 vs 14.9 +/- 3.6 cm/s; p < 0.05; ADPV 27.6 +/- 5.2 vs 19.7 +/- 4.7 cm/s; p < 0.05), and a decrease in the systolic index ASVP (3.8 +/- 1.3 vs 7.6 +/- 2.6 cm/s; p < 0.05). The diastolic indices recorded with IABP did not change in Group 2. The velocity spectra changed with the appearance of abnormalities usually described in the presence of microcirculatory abnormalities ("no reflex" phenomenon): decrease in anterograde systolic flow, rapid deceleration of diastolic velocities with appearance of a retrograde systolic flow. The authors conclude that IABP increases diastolic velocities of coronary flow in the acute phase of revascularised anterior myocardial infarction complicated by left ventricular failure but does not seem to be accompanied by improved myocardial perfusion. PMID- 12741309 TI - [Cardiovascular abnormalities of the antiphospholipid antibody syndrome]. AB - The antiphospholipid antibody syndrome (APS) may present with serious cardiovascular complications which should be recognised by the cardiologist. The authors report a series of 6 cases of APS diagnosed after thrombotic events and the finding of antiphospholipid antibodies. The APS was primary in 5 cases and associated with tuberculous lymphadenitis in 1 case. There was cardiac involvement in 5 patients with pericardial effusion in 3 cases, complicated by tamponade as the presenting sign of primary APS in the other 2, valvular disease in one case (moderate mitral stenosis with aortic valve disease) and pulmonary embolism in one case. Five patients developed recurrent deep vein thrombosis of the legs. One patient had a transient ischaemic cerebral attack. PMID- 12741311 TI - [Isolated non-compaction of the left ventricle]. AB - Isolated non compaction of the left ventricle is a rare congenital cardiomyopathy linked to an arrest of normal myocardial embryogenesis. We report two cases of isolated non compaction of the left ventricle discovered by echocardiography in 2 males of 30 and 55 years. The first had progressively worsening cardiac insufficiency, the second was being followed for an unexplained cardiomyopathy. In both cases, the diagnosis was able to be confirmed by transthoracic echocardiography, supported by MRI data. Although present from birth, this condition can become apparent at various ages and is complicated by sudden death (principal cause of mortality), severe cardiac insufficiency, or thrombo-embolic accidents. The diagnosis of left ventricular non compaction should be considered when faced with unexplained cardiac insufficiency in the adult. PMID- 12741310 TI - [Electromechanical mapping of myocardial ischemia in coronary occlusion in the pig]. AB - The NOGA-Biosense catheter-based mapping technique has been well studied experimentally in infarction model. However, chronic myocardial ischemia with this new device has not been well explored. Thus, the aim of our study was to assess electromechanical changes in a pig aneroid constricor model. To achieved this aim, ten pigs were studied 21 days after the implantation of an aneroid constrictor around the circumflex artery. Coronary reserve assess by intracoronary Doppler flow wire was reduced in the ischemic lateral area (ILA) compared with the nonischemic zone (NIZ) (1.3 +/- 0.1 in the ILA vs. 2.3 +/- 0.2 in the NSZ; p < 0.01). TM echocardiography was used to evaluate myocardial regional contractility under basal condition and after stress induced by rapid atrial pacing. In stress state, the ischemic zone showed an impaired contractility compared with basal state (wall thickening, 32.7 +/- 7.4% vs. 59.7 +/- 8.6%; p < 0.05) whereas the non ischemic zone did not (53.8 +/- 7.6% vs. 60.8 +/- 10.1%; p = ns). Constrast echography showed a decrease in contrast intensity in subendocardium of the ila compared with the niz (46.2 +/- 16.6 vs. 99.2 +/- 35.6; p = 0.03) in pacing. Ventricular mapping quantified unipolar (UV). bipolar (BV) voltage potentials and endocardial local shortening (LLS) in 9 left ventricular regions. In basal state, electrical potentials were preserved in both zones (UV: 9.1 +/- 1.8 mV in the ischemic vs 11.3 +/- 3.6 mV in the non ischemic zone; p = ns; BV: 4.2 +/- 1.1 mV in the ILA vs. 3.9 +/- 1.5 mV; p = ns). In contrast, LLS was significantly lower in the ischemic compared with non ischemic zone (6.4 +/- 5.4% vs. 17.9 +/- 3.0%, p < 0.001). In conclusion, ventricular mapping with the NOGA-Biosense system can identify the ischemic myocardium. In this pig model, the association of a preserved electrical activity and an impaired mechanical activity characterizes the ischemic myocardium. These findings could be interesting in this model in regard of the new developments of the system in particular in the field of angiogenesis. PMID- 12741312 TI - [An exceptional etiology of left ventricular aneurysm: type AA amyloidosis]. AB - Left ventricular aneurysms most often occur in the course of myocardial infarction. In rare cases they can be detected when the coronary network is devoid of any lesions. The aetiology is therefore multiple and dependent on the context. One aetiology seems less exceptional and concerns idiopathic aneurysms encountered in the African population, where the role of a "debilitating condition" such as tuberculosis has been evoked. We report the case history of a young patient from Zaire with a left ventricular aneurysm discovered in association with ganglionic tuberculosis complicated by AA amyloidosis. Histological analysis allowed the aetiological diagnosis to be established. Aneurysmal dilatation of the left ventricle was reported in the presence of amyloid deposits at the intra-myocardial arteriole level, whereas the context suggested a tubercular role. In spite of the difficulty of establishing a precise aetiological diagnosis, there seems to exist a consensus for surgical management. PMID- 12741313 TI - ["Spontaneous" rupture of the left iliac vein complicating Cockett's syndrome]. AB - The case history reported concerns a female patient aged 42 years for whom the clinical picture was that of a blue phlebitis (phlegmatia caerulea dolens), associated with a state of shock evoking a severe pulmonary embolus. The absence of echocardiographic dilatation of the right cavities, and the appearance of a left iliac fossa mass, steered the diagnosis towards internal haemorrhage. Emergency laparotomy allowed diagnosis and treatment of a so-called spontaneous rupture of the left iliac vein, a rare condition for which 20 cases have been reported in the literature. Re-operation performed 24 hours afterwards for the absence of venous return allowed the discovery of Cockett's syndrome with ascending thrombosis, requiring cross-venous bypass associated with the creation of an arterio-venous fistula in order to maintain permeability. One year afterwards the appearance of signs of cardiac insufficiency led to the closure of this fistula. PMID- 12741314 TI - [Primary pericardial sarcoma]. AB - We present the case of a 50 years old male revealed by a recurrent pericardial effusion. The diagnosis of malignancy was confirmed by direct biopsy. The treatment consisted in surgical excision and chemotherapy. The patient was asymptomatic 17 months after surgery. PMID- 12741315 TI - [Screening of microbial secondary metabolites inhibiting cholesterol biosynthesis with the use of hepatoblastoma G2]. AB - The culture of hepatoblastoma G2 (Hep G2) cells is proposed as an effective model for screening of microbial metabolites--inhibitors of sterol biosynthesis. This model can be applied at early stages of screening procedures and is quite effective for testing of crude extracts of producers' culture broth. The test is based on measurement inhibition of the radiolabelled precursors incorporation in cholesterol and separate fractions of lipids by microbial metabolites in Hep G2 cells. That allows not only to reveal inhibitors of cholesterol biosynthesis, but also to evaluate mechanism of action, including ability to inhibit the synthesis of cholesterol ethers. The cholesterol biosynthesis inhibition was tested at 150 microbial cultures (actinomycetes and imperfect fungi), isolated from soil. The ability to inhibit 14C-acetate incorporation into cholesterol was found in 15-20% of microbial cultures possessing antifungal activity of extracts (culture broth and mycelium). PMID- 12741316 TI - [Antiherpetic activity of l-lysine-alpha-oxidase in different dosage forms]. AB - The antiviral effect of different medicinal forms on base of L-lysin-alpha oxidase (LO) in treatment of experimental keratits and skin diseases induced by herpes simplex virus type 1 (HSV-1) at the rabbits was examined. Treatment with gel form of LO was more effective in comparison with water solution form of LO. This effect was tested by morphological, histological and immunoblotting methods in dynamics of viral disease. PMID- 12741317 TI - [Cell wall components of gramicidin S resistant Staphylococcus aureus]. AB - Comparative study of two staphylococcus aureus 209P strains--resistant and susceptible to gramicidin S demonstrated that peptidoglycanes of two strains differ by ratio glycine/serine at peptide bridges. Besides peptidoglycanes significantly differ by amidation of alfa-carboxyles of glutamic acid in muropeptide. This peptidoglycane modification of resistant cells along with enhanced content of etherized D-alanine in teichoic acid provides lower negative charge of cell wall components. It may influence the cell wall ability to react with positively charged gramicidin molecules. It was shown that isolated cell walls and peptidoglycane of resistant cells binds significantly less gramicidin than cell walls and peptodoglyce of susceptable cells. Simultaneous determination of gramicidin binding by intact S. aureus cells and their killing revealed that lower ability of resistant cells to bind gramicidin is significant but not critical factor of gramicidin resistance. PMID- 12741318 TI - [Antimicrobial activity of Laetiporus sulphureus strains grown in submerged culture]. AB - Cultural conditions for growth and fruit body formation were elaborated to four strains of Laetiporus sulphureus isolated from nature. All strains demonstrated antimicrobial activity against a wide spectrum of gram-positive and gram-negative bacteria during agar and submerged cultivation including methicillin-resistant strain of Staphylococcus aureus (MRSA) and glycopeptide-resistant strain of Leuconostoc mesenteroides. Antifungal activity was not found. The level of antimicrobial activity during submerged cultivation reached maximum after seven days of growth on specific medium with soybean meal and corn liquid; the next four weeks its increasing was not so manifested. Antimicrobial activity correlated with orange pigment secretion and cultural liquid acidification to pH 2.0-2.8 that indicates on acid nature of synthesized products. PMID- 12741319 TI - [Enhanced activity of rifampicin loaded with polybutyl cyanoacrylate nanoparticles in relation to intracellularly localized bacteria]. AB - Association of rifampicin with polybutylcyanoacrylate nanoparticles provided considerable enhancement of drug antibacterial activity. In vitro nanoparticle loaded rifampicin was more active against Staphylococcus aureus and Mycobacterium avium, localized in isolated alveolar macrophages. Level of rifampicin in macrophages increased 2-3-fold after incubation with rifampicinloaded nanoparticles comparing to the free drug. High therapeutic efficacy of colloidal rifampicin was demonstrated in vivo. Use of nanoparticles provided 2-fold increase in rifampicin efficacy, comparing with the free drug at treatment of staphylococcus sepsis in mice. Single administration of nanoparticulate rifampicin in the dose 25 mg/kg resulted in 80% survival of mice with salmonellosis, while 50 mg/kg of free rifampicin could provide only 10% survival. It may be considered that high antibacterial efficacy of rifampicin bound to nanoparticles is due to its effective delivery to macrophages. PMID- 12741320 TI - [Microbiological evaluation of differences between cephalosporins of second and third generations in general hospital]. AB - The aim of the current investigation was to evaluate practical impact of modern NCCLS recommendations for the selection of 2nd and 3rd generation cephalosporins in Moscow teaching multi profile hospital. The sensitivity of clinically significant 96 strains from patients with pyelonephritis and 180 strains from patients with lower respiratory tract infections (pneumonia, COPD) was compared for cefuroxime and cefotaxime or ceftriaxone according NCCLS recommendations during 2000-2001 years. At the lower respiratory tract infection total sensitivity of all pathogens was 70.6% and 72.8%, at the pyelonephritis 71.9% and 76.0% for 2nd and 3rd generations respectively. The differences between cephalosporins were not statistically significant. Based on the application of modern NCCLS recommendations in the routine microbiological practice similar clinical efficacy of 2nd and 3rd generations cephalosporin in lower respiratory tract infections and pyelonephritis could be predicted. PMID- 12741321 TI - [Cytoreductive surgery with intraperitoneal chemotherapy in patients with colon cancer and peritoneal carcinomatosis]. AB - The efficacy of cytoreductive approach for colon cancer with carcinomatosis was assessed. 34 patients (the main group) underwent colon resections, peritonectomy, omentectomy and intraperitoneal chemotherapy (mitomycin and 5-fluorouracil). 22 patients (control group) underwent palliative colon resections only. The spread of peritoneal dissemination was assessed in all patients, basing on Peritoneal Cancer Index (PCI). It was found, that the rate of postoperative morbidity is higher in the main group, but it was not accompanied by enhanced postoperative mortality. It was also detected, that cytoreductive approach permits to prolong the survival (17.0 +/- 3.5 months in main group vs. 7.5 +/- 2.1 months in controls). The main prognostic factor in these patients was the PCI. The most favorable prognosis is identified if PCI < 5 (few discrete implants are present); the survival in this group was 23.0 +/- 4.5 months and disease-free interval was 14.0 +/- 2.5 months. When PCI was higher then 5, the survival was 14.5 +/- 2.5 months and disease-free interval was 7.0 +/- 1.3 months. PMID- 12741322 TI - [Bacteriophages for treatment and prophylaxis of infectious diseases]. PMID- 12741323 TI - [Molecular and cellular mechanisms of fluoroquinolones immunomodulating activity]. PMID- 12741325 TI - [Electromagnetic interferences and cardiac stimulation and in defibrillation: from the aspect of the specialist]. PMID- 12741324 TI - [A scientific journey: electromagnetic fields, cardiac pacemakers and defibrillators]. PMID- 12741326 TI - [Exposure levels in the environment]. AB - Presentation of electromagnetic sources constituted by various radio transmitters contributing to different radio communication services in the environment. Results of a campaign of measures to assess the electromagnetic field in the close neighborhood of various stations. Analysis by frequency domains. PMID- 12741327 TI - [Interferences in the everyday life of the patient with a cardiac pacemaker or an implantable defibrillator]. AB - The electromagnetic sources of interferences being able to deteriorate the operation of cardiac pacemakers or implantable defibrillators are numerous. This potential risk has been known since the release, 40 years ago, of pacemakers incorporating a detection circuit. Many papers, reviewed in this article, have been published about these conflicts. In daily practice, the risk of dangerous interference is weak, but it seems obvious that the implantable defibrillators are much more sensitive to the external environment than the cardiac pacemakers. With some precautions to eliminate manifest risk situation, it is possible to carry out a strictly normal life. Diagnostic memories increasingly sophisticated included in the new prostheses make possible the identification of asymptomatic conflicts, and the specification of the mechanism of a real problem. Provided information may also be useful to find solutions (adjustment, precautions) to decrease, even to remove the risks. PMID- 12741328 TI - [Relevance of in vitro studies for the immunity of cardiac implants in an electromagnetic field environment]]. AB - The object of this article is to show the contribution of the vitro experimental approach, based on the basic rules of electromagnetic compatibility (EMC), as a complementary tool to the clinical studies. Results can be obtained by a experimental approach in vitro with electromagnetic phantoms associated to a bench of "standard" test to allow a possible comparison between various studies. After describing the protocol developed for the cardiac implants (pacemakers and defibrillators), examples of partial results are presented by way of illustration. PMID- 12741329 TI - The interference of electronic implants in low frequency electromagnetic fields. AB - Electronic implants such as cardiac pacemakers or nerve stimulators can be impaired in different ways by amplitude-modulated and even continuous electric or magnetic fields of strong field intensities. For the implant bearer, possible consequences of a temporary electromagnetic interference may range from a harmless impairment of his well-being to a perilous predicament. Electromagnetic interferences in all types of implants cannot be covered here due to their various locations in the body and their different sensing systems. Therefore, this presentation focuses exemplarily on the most frequently used implant, the cardiac pacemaker. In case of an electromagnetic interference the cardiac pacemaker reacts by switching to inhibition mode or to fast asynchronous pacing. At a higher disturbance voltage on the input of the pacemaker, a regular asynchronous pacing is likely to arise. In particular, the first-named interference could be highly dangerous for the pacemaker patient. The interference threshold of cardiac pacemakers depends in a complex way on a number of different factors such as: electromagnetic immunity and adjustment of the pacemaker, the composition of the applied low-frequency fields (only electric or magnetic fields or combinations of both), their frequencies and modulations, the type of pacemaker system (bipolar, unipolar) and its location in the body, as well as the body size and orientation in the field, and last but not least, certain physiological conditions of the patient (e.g. inhalation, exhalation). In extensive laboratory studies we have investigated the interference mechanisms in more than 100 cardiac pacemakers (older types as well as current models) and the resulting worst-case conditions for pacemaker patients in low-frequency electric and magnetic fields. The verification of these results in different practical everyday-life situations, e.g. in the fields of high-voltage overhead lines or those of electronic article surveillance systems is currently in progress. In case of the vertically-oriented electric 50 Hz fields preliminary results show that per 1 kV/m unimpaired electrical field strength (rms) an interference voltage of about 400 microVpp as worst-case could occur at the input of a unipolar ventricularly controlled, left-pectorally implanted cardiac pacemaker. Thus, already a field strength above ca. 5 kV/m could cause an interference with an implanted pacemaker. The magnetic fields induces an electric disturbance voltage at the input of the pacemaker. The body and the pacemaker system compose several induction loops, whose induced voltages rates add or subtract. The effective area of one representing inductive loop ranges from 100 to 221 cm2. For the unfavourable left-pectorally implantated and atrially-controlled pacemaker with a low interference threshold, the interference threshold ranges between 552 and 16 microT (rms) for magnetic fields at frequencies between 10 and 250 Hz. On this basis the occurrence of interferences with implanted pacemakers is possible in everyday-life situations. But experiments demonstrate a low probability of interference of cardiac pacemakers in practical situations. This apparent contradiction can be explained by a very small band of inhibition in most pacemakers and, in comparison with the worst-case, deviating conditions. PMID- 12741331 TI - [Interferences and cardiac pacemakers--defibrillators. Results of in vivo experiments and radio frequencies]. AB - Interference with cardiac pacemakers and defibrillators by cellular phone and electronic article surveillance systems is shown in experimental studies with disparate findings. Interaction occurrence in real life is a convincing but rare experience. Device model, distance, power output and technology of the source are different and sometimes uncontrollable factors. As a result it remains difficult to quantify the true incidence of interaction and associated health risk. Nevertheless, simple recommendations commonly help the patients to prevent the interference. PMID- 12741330 TI - [Effects of 50 to 60 Hz and of 20 to 50 kHz magnetic fields on the operation of implanted cardiac pacemakers]. AB - The effect of 50 Hz and 60 Hz (frequencies of current distribution) and 20 kHz to 50 kHz (frequencies of induction cooktop) magnetic interference on implanted pacemakers have been assessed with the present generation of device technology. Sixty patients implanted in 1998 and 1999 with dual chamber pacemakers from 9 different manufacturers were monitored with telemetry while passing through, and standing between a system of two coils. They generated a 50 Hz or a 60 Hz magnetic field at 50 microT. Then, patients used a cooktop at different power. The recordings were made with the standard setting of "medically correct" sensing parameters chosen for the patients. Then pacemakers were reprogrammed to the unipolar mode, with the highest atrial (A) and ventricular (V) sensitivity that did not induce muscular inhibition while moving. Between each exposure (50 Hz, 60 Hz or 20 kHz to 50 kHz), the pacemaker programmation was controlled. At the end of the tests, pacemakers will be reprogrammed with the standard setting. The medical observer being blind to the existence or not of the magnetic field. No pacemaker was influenced by the vicinity of the magnetic field at medically correct settings. At unipolar high sensitivity, no inhibition nor reprogramming was observed. Transient reversion to interference mode was observed in 6 cases, 3 transient acceleration due to atrial detection of the interference, and one T wave detection by the ventricular lead. All were observed with the 60 Hz, and only 3 with the 50 Hz magnetic field. One device (Biotronik) shifted out of its special program (hysteresis research) during the tests with the induction cooktop, but it maintained its standard program, and the event could not be repeated despite further testing. CONCLUSION: Actual pacemakers do not present any electromagnetic interference with 50 Hz and 60 Hz or induction cooktop frequency working. They are insensitive with medically correct settings. Unusual high sensitivity leads only to noise reversion mode, or transient ventricular tracking. PMID- 12741332 TI - [Cardiac Pacemakers, implantable defibrillators and IRM]. AB - The IRM is formally contraindicated to the pacemaker and cardiac defibrillator wearers because of the risk of inhibition or inappropriate stimulations during the examination. However if the examination is essential, suitable programming of the apparatus and a constant monitoring of the heartbeat rate by a qualified doctor in cardiac stimulation must make it possible to avoid any accident. PMID- 12741333 TI - [Normative aspects of sources]. AB - The human body is more and more exposed to the electromagnetic phenomena generated by all domestic and professional electric appliances. This problem "EMF and health" is devoted to the possible effects known as "biological" and the possibilities of disturbances of the medical implants. Although many studies still did not give conclusions on these long-term effects, international standardisation is in progress and could lead to a difficult use of electricity such that it currently exists. The modelling of the phenomena in the human body can contribute to a better knowledge of the interactions field-man and to the development of more realistic standards. PMID- 12741334 TI - [Medical implantable devices and electromagnetic compatibility]. AB - Continuing progress in medicine has led to a corresponding population growth among the elderly population resulting in an increase in the number of patients with active implanted medical devices. At the same time, there continues to be a proliferation of electromagnetic wave sources within our technological environment. The coexistence of implanted active medical devices and environmental electromagnetic waves requires particular attention in order to avoid electromagnetic interference. For this reason, experts are more and more involved in writing specific manufacturing standards. PMID- 12741335 TI - [Compatibility of active implants in the professional environment]. AB - The recommended limiting values of human exposure are much higher than the thresholds of immunity required by the standards of electromagnetic compatibility. It is thus impossible to guarantee the total absence of interference risk. So initially, the medico-legal responsibility suggested proscribing the aptitude for a potential exposure, the importance of psychosocial problems raised by such a step forces to better quantify this risk which appears altogether weak with the collective experiment. The principal sources of professional exposure and the levels of corresponding fields are presented, as well as current knowledge on the sensitivity of pacemakers and defibrillators with respect to these fields. Finally, information to be supplemented and the means of mitigating it are suggested. PMID- 12741336 TI - [Choice criteria and complications of urinary diversions]. AB - The choice of urinary diversion is conditioned to patient's disease, performance status, age and life style. Ureterointestinal anastomosis is a critical stage in urinary diversion, allowing urinary transit and preventing reflux. We have examined urinary diversion frequently used in our clinical practice. In ureterosigmoidostomy and MAINZ pouch II , ureterointestinal anastomosis isn't refluent. Ileal conduit, reserved to patient with advanced disease and/or low life expectation, normally the implantation is direct. In continent reservoir and orthotopic neobladder, detubularization produces low pressure. In these urinary diversion anti-reflux anastomosis isn't mandatory, because the risk of stenosis is higher. Urinary infection is an important criterion in choice of anastomosis. After all is emphasized that success of ureterointestinal implantation doesn't depend on surgeon's level of experience. PMID- 12741337 TI - [Ileal conduit and urinary stoma complications]. AB - The ileal conduit has been widely used for urinary diversion. It is a safe procedure with acceptable results, but it has become clear that the conduit can give rise to serious complications, notably renal deterioration in the long run. Stenosis of the ileal conduit, usually developing insidiously many years after the diversion, may be the cause of upper urinary tract damage. In most cases, a variety of possible factors is considered, including microvascular ischemia, urine-borne toxic material, infectious and allergic stimuli and an immunologic defect. Crohn's disease may affect an ileal loop urinary conduit. It presents as a diffuse loop stenosis. Recurrent transitional carcinoma arising within an ileal conduit following cystectomy for malignant disease is rare, but late malignancy in bowel segments exposed to urine without fecal stream is well known. In literature, distinction between conduit complications of patients with underlying benign disease and those with malignancy has not always been clear. Stoma and skin complications are frequently observed and the patients who practiced inadequate stoma care routines are more likely to show peristomal skin complications. Patients bearing an abdominal urostomy should be followed up stringently in stoma centers. PMID- 12741338 TI - [Complications of urinary diversion after radiotherapy]. AB - Primary radiation therapy may be recommended for patients with invasive bladder cancer, gynecological or prostatic cancer. When complications occur or in case of malignant recurrence, urinary diversion may be the best chance to restore an acceptable quality of life. The complication rate after this surgery is doubled. We report our experience in 32 patients submitted to urinary diversion after radiotherapy from 1985 to 2000: 2 enteric fistulas; 2 urinary fistulas; 5 stenosis of uretero-intestinal anastomosis were our complications. Radical cystectomy (24 cases) or anterior pelvic exenteration (8 females) preceded urinary diversion. Preoperative high-dose radiotherapy contributes to increased postoperative morbidity rates, particularly entero-enteric fistulas, uro intestinal fistulas and stenosis of the uretero-intestinal anastomosis. In our experience, in most of the major urinary or enteric complications non surgical management was inefficient and surgical management was necessary. PMID- 12741339 TI - [Metabolic complications of urinary intestinal diversion]. AB - The use of bowel in urinary diversion has increased markedly over the last 15 years and this trend is expected to continue. Complications that have been noted in patients with urinary intestinal diversion occurring as a consequence of metabolic abnormalities include disordered electrolyte metabolism, osteomalacia, growth retardation, calculus formation, infection, abnormal drug metabolism. The factors that influence the intensity of absorption of urinary electrolites by intestinal mucosa include length of time of retention of the urine, concentration of solute in the urine, type of urinary diversion, quality of emptying, capacity of the reservoir, surface area of the bowel used, renal function, infection and chronic dilatation. Regular draining of the urinary intestinal diversion, vigilant metabolic follow-up, and careful patient selection are essential to prevent metabolic abnormalities. PMID- 12741340 TI - The start of pharmacological activity after sublingual administration of sildenafil citrate in 30 patients affected by erectile dysfunction. AB - INTRODUCTION AND OBJECTIVES: Sildnenafil citrate is a powerful phosphodiesterase type 5 isoenzyme; it is the first oral treatment to have had a significant success in treatment of erectile dysfunction (ED). After oral dosing on an empty stomach the pharmacological activity starts within 30 to 120 minutes (average 60 minutes) whereas the effect of this medication after a meal could be notably delayed. We evaluated the start of pharmacological activity in 30 patients affected by non-psychogenic ED after sublingual administration of Sildenafil citrate. METHODS: Patients participating in our study were all affected by ED whose etiology was assessed as vasculogenetic or diabetic. The study lasted 6 months. For the first 3 months patients were asked to take Sildenafil (50-100 mg) for oral administration, under normal everyday conditions, 30 minutes before planned sexual relations. During the second 3 months the patients were asked to take Sildenafil for sublingual administration (crushing the pill in the mouth and dissolving the drug under the tongue) 15 minutes before planned sexual relations. The patients did not know the purpose of the study. RESULTS: An appreciable reduction in the start of pharmacological activity was reported during the time of sublingual administration. In fact, while throughout the first 3 months the average pharmacological onset was 62.8 minutes (DS +/- 16.8), during the second 3 months it was 29.3 minutes (DS +/- 8.1), the mean difference in the start of pharmacological activity was 35.3 (DS +/- 12.4). The results of T-student test for paired observation were T (29) = 15.629; p-value 0.00001. During the two modalities of administration no differences were noted in the efficacy or in the frequency of adverse events. All the patients declared they preferred the sublingual way because of faster onset. CONCLUSIONS: Even though ours is a limited study, our clinical data points out that the sublingual administration of Sildenafil is useful because of the rapid onset unrelated to meals. All the patients were reported to appreciate this method of administration, particularly in the case of unplanned sexual relations. PMID- 12741341 TI - Bilateral varicocele: single transscrotal approach for Tauber antegrade sclerotherapy. AB - OBJECTIVE: To describe the results of a new simple technique of single transscrotal approach for Tauber antegrade sclerotherapy in bilateral varicocele. MATERIALS AND METHODS: During the period March 1998-June 2001, overall 341 patients were treated for varicocele using Tauber antegrade sclerotherapy. In 39 of these patients, sclerotherapy was performed bilaterally in the same session whilst in 34, rather than making two incisions at the root of the two hemiscrotums, a single incision was made on the median raphe. During this period, the same approach was used in 4 patients with left varicocele and right hydrocele, 2 patients with left varicocele and right epididymis cyst, 1 patient with bilateral varicocele and bilateral hydrocele, 1 patient with bilateral hydrocele and left varicocele. RESULTS: Maximum ray exposure time was 58 seconds (mean 30). Oro-tracheal intubation was not necessary in any of the patients. In 12 patients, besides local anesthesia, additional sedation was given. Slight bleeding of the wound, occurring in 3 patients, was medicated in the outpatient department. CONCLUSIONS: Patients prefer a single incision. When the incision is made on the median raphe, no scars remain. In bilateral varicocele, the single approach reduces invasiveness and increases patient satisfaction. PMID- 12741342 TI - A case report of CUAVD with azoospermia: a proposal of a rational diagnostic approach. AB - Since its introduction in 1992, intracytoplasmic sperm injection (ICSI) has made the treatment of severe male infertility possible, particularly that of azoospermia, both secretory and secretory. Some azoospermic subjects have a pathological development of the seminal pathways, and in particular of the vas deferens and/or ejaculatory ducts. A large part of these subjects show, like patients affected by cystic fibrosis, mutations in the cystic fibrosis transmembrane regulator (CFTR) gene. Some of these azoospermic subjects are indeed paucisymptomatic fibrosis cystic patients who bear the risk of transmitting cystic fibrosis, seminal pathways alterations and, possibly, renal malformations to their offspring. We describe a case of an infertile patient with right CUAVD and azoospermia previously treated by crossed epididimovasostomy in the absence of any genetic and an adequate anatomic evaluation. He was then found to be CFTR mutation positive and without demonstrable spermatozoa in the vesicular fluid despite ultrasound evidence of left ejaculatory duct obstruction. During the second TESA-ICSI cycle an ongoing pregnancy was obtained. An extensive genetic examination for CFTR mutations and a through anatomical study is, therefore, mandatory in these patients to select the most appropriate treatment in CFTR mutation positive and negative CUAVD patients. PMID- 12741343 TI - [Sperm characteristics in fertile men and healthy men of the south-east Sicily from year 1982 to 1999]. AB - OBJECTIVE: To investigate whether sperm density (D) and morphology (M) deteriorated in the last 18 years (from 1982 to 1999) in a South-East area of Sicily (Catania, Siracusa and Ragusa provinces). The area of residence of this subjects studied is a strong consumer in fumigants and pesticides in the hot house factories and thus the present study can be useful to examine a possible negative impact of these compounds on sperm parameters through a long-term monitoring of an apparently healthy population. MATERIALS AND METHODS: Sperm D and M were retrospectively analysed in 716 men (median age 29, range 22-43 years) referring to the Andrology Center of the University of Catania. The subjects were subdivided in 2 groups: the first included subjects who fathered occasionally (n = 467), the second was formed by healthy volunteers consulting as possible donors (n = 109) and to check their fertility (n = 140). RESULTS: During the study period of 18 years, both sperm D and M (in terms of percentage of normal forms) showed significant (p < 0.01) time-dependent indirect correlations (r = -0.12 for D and r = -0.29 for NF). D exhibited two significant peak values in the years 1982, 1983 and 1994 respectively, followed by two periods (years 1984-1986, and 1995-1996) with not significantly declined values and then with values significantly lower than the above peak values in the years 1987, 1989, 1992 as well as in the period 1997-1999. The M registered a progressive decline, statistically significant in the years 1988 and 1994 in comparison to the preceding years. A further significant decrease in sperm M was registered, in comparison to the periods 1982-1987 and 1988-1994, in the period 1995-1999. CONCLUSION: Our data showed a temporal decline of sperm parameters in South-East area of the Sicilia, from 1982 to 1999, in fertile men and occasional healthy subjects or donors. These data confirmed the sperm deterioration registered in Europe. The results might be influenced by time-varying body of users of referring Andrology Center (i.e. oligozoospermic subjects preferring ART program have approached again our Center in the last years from the activation of our Unit of ART), and/or by varying temporal changes in the levels of environmental/occupational gonadotoxic factors. PMID- 12741344 TI - [Varicocele and coincidental abacterial prostato-vesiculitis: negative role about the sperm output]. AB - OBJECTIVE: To evaluate the frequency and the role of a coincidentally expressed abacterial prostato-vesiculitis (PV) on sperm output in patients with left varicocele (Vr). MATERIALS AND METHODS: We evaluated 143 selected infertile patients (mean age 27 years, range 21-43), with oligo- and/or astheno- and/or teratozoospermia (OAT) subdivided in two groups. Group A included 76 patients with previous varicocelectomy and persistent OAT. Group B included 67 infertile patients (mean age 26 years, range 21-37) with OAT and not varicocelectomized. Patients with Vr and coincidental didymo-epididymal ultrasound (US) abnormalities were excluded from the study. Following rectal prostato-vesicular ultrasonography, each group was subdivided in two subsets on the basis of the absence (group A: subset Vr-/PV-; and group B: subset Vr+/PV-) or the presence of an abacterial PV (group A: subset Vr-/PV+; group B: subset Vr+/PV+). Particularly, PV was present in 47.4% and 41.8% patients of groups A and B, respectively. This coincidental pathology was ipsilateral with Vr in the 61% of the cases. Semen analysis was performed in all patients. RESULTS: Patients of group A showed a total sperm number significantly higher than those found in group B. In presence of PV, sperm parameters were not significantly different between matched--subsets (Vr-/PV+ vs. Vr+/PV+). In absence of PV, the sperm density, the total sperm number and the percentage of forward motility from subset with previous varicocelectomy (Vr-/PV) exhibited values significantly higher than those found in the matched--subset (Vr+/PV-). CONCLUSION: Sperm analysis alone performed in patients with left Vr is not a useful prognostic post varicocelectomy marker. Since following varicocelectomy a lack of sperm response could mask another coincidental pathology, the identification through US scans of a possible PV may be mandatory. On the other hand, an integrated uro-andrological approach, including US scans, allows to enucleate subsets of patients with Vr alone, who will have an expected better sperm response following Vr repair. PMID- 12741345 TI - [Questionnaire based evaluation of prostate biopsy complication comparing different bioptic schemes]. AB - OBJECTIVES: The systematic parasagittal sextant biopsy technique under transrectal ultrasound guidance, routinely performed to diagnose and stage prostate cancer, has been shown to outperform directed or random biopsies, revolutioning our ability to detect carcinoma of the prostate. Different biopsy schemes have been proposed with similar positive percentage of cancer detection. The present study evaluate from the patient's perspective the complications with the use of two different systematic biopsy protocols with 14 and 8 cores. MATERIAL AND METHODS: Between January 1999 and February 2000, 177 consecutive patients, mean age 64.1 +/- 7.7 years, referred for normal screening digital rectal examination (DRE) and prostate specific antigen (PSA) level 4-10 ng/ml, were submitted to a transrectal US examination followed by lesion directed and 14 scheme systematic biopsies to detect prostate cancer. Biopsies were obtained from conventional sextant biopsies (6 core) and 3 alternate sites which included: the right and left extreme lateral peripheral zone between anterior tissue and posterior gland base (2 core); the right and left transition zone, immediately adjacent to the urethra anterior and posterior (4 core) and the right and left central gland in the mid zone typical of benign prostatic hyperplasia (BPH) (2 core). All specimens were separated for specific location identification. Cancer was identified in 61 patients (34.46%). Traditional sextant biopsies showed 23 patients (37.7%) with positive core to detect cancer, while a sextant regimen incorporating lateral peripheral zone biopsies and transitional zone detected 19 cancer (31.1%). The combination of lateral peripheral and transitional zone alone detected cancer in 59 patients. No cancer was detected in central gland. The lateral peripheral zone was the most frequently positive site biopsy followed by the transitional zone. According the results of our study from April 2000 we started to consider a novel scheme to reduce number of biopsies maintaining the same sensitivity. A subsequent group of 121 patients, mean age 61 +/- 4.6 years, enrolled from April 2000 to May 2001, underwent a transrectal US examination followed by lesion directed and 8 scheme systematic biopsies. None of the patients had previously undergone prostate biopsy. In all patients a visual analog score (VAS) questionnaire about pain and complications was obtained 7 days after the procedure. RESULTS: Of the 149 patients who completed the questionnaire 9.9% found the procedure moderately to extremely painful afterwards, with a VAS > 5, the commonest of these complications being pain and voiding difficulties with a mean value of 15.8%, systemic symptoms as fever or sweats with a mean value of 7.65% of cases. Between the two groups submitted to 14 or 8 scheme biopsies, we detect a statistically significant difference for urethral bleeding (7.3% vs 4.9% p value 0.05) and rectal bleeding (10.3% vs 3.7% p value 0.04), systemic symptoms 10.3% vs 5.0% p value 0.05) and painful voiding afterwards (5.8% vs 2.4% p value 0.02). CONCLUSIONS: Ultrasound guided transrectal biopsy of the prostate is a well tolerated and effective method for obtaining multiple biopsy specimens from the prostate with low incidence of serious complications. The absolute value of referred complications with VAS < 5 results high, but the rate of major complications results low. The 8 biopsy scheme, including sampling in peripherial zone at midgland and transition zone periuretrally toward the base, should be considered in a initial biopsy scheme to reduce number of biopsy and enhancing sensitivity, with a significant less degree of complication rate compared to a extensive 14 biopsy scheme. PMID- 12741346 TI - [The meaning and usefulness of spiral CT for radiolucent ureteric stones diagnosis: our experience]. AB - The aim of this work was to report some case histories on the usefulness of spiral TC, used for several years both to diagnose renal colic and urinary lithiasis and to study radio lucent stones that are often difficult to be detected with traditional radiology. 13 patients, aged between 31 and 76 (average age: 54.2), were therefore examined. Eight of them had a ureteral colic when examined, while five patients had shown symptoms some days before being hospitalised in our ward. In all cases, ultrasonography showed a significant hydronephrosis, while direct radiography of the urinary tract could not detect any images that could be associated with radio-opaque lithiasis. All patients therefore underwent an abdominal spiral TC with no contrast medium within 24 hours after hospitalisation. The confrontation between the results obtained by ultrasonography and those obtained by spiral TC, showed the usefulness of the former method to detect stones located in the proximal ureter or in its intramural tract, while the latter could detect the lithiasis of the proximal ureter in 3 cases (23%), of the mid ureter in 2 cases (15.3%), and of the distal ureter in 8 cases (61%). The stones had, approximately, a 5 mm diameter in 5 cases. In 6 cases the diameter was between 6 and 10 mm, and more than 1 cm in 2 cases. Both methods proved to be equally accurate in the assessment of the hydronephrosis degree and of the thickness of the renal parenchyma. The therapy was medical in 2 cases and open surgery in 3 cases, while 8 patients were treated with ureterolitholapaxy with a ballistic searcher. The usefulness of TC in the study of urolithiasis nowadays is supported by a large literature which clearly supplies with documentary evidence the high sensitivity and specificity of such a method in diagnosing the presence of urolithiasis in general and above all of ureteric stones. Such a method not only makes an accurate evaluation of the stones location possible, but it can also assess the calculi dimensions and the indirect signs of the functionality of the kidney affected, without having to use the contrast medium. This method needs very limited execution times and allows a diagnostic of possible collateral pathologies. The main disadvantage of spiral TC, if compared to conventional radiology, is that the patient is exposed to a larger quantity of ionizing radiations, although such an inconvenience will be overcome by the new and more technologically advanced machines. According to our experience, though based on a limited number of cases, spiral TC allowed us to get a quick diagnosis of radio-lucent lithiasis, to see the seat and dimensions of the calculi and finally to chose the most effective treatment. We can therefore think of a diagnostic protocol, for ureteral colics with hydronephrosis or complicated by hyperpyrexia or sepsis, with spiral TC in order to have a quick diagnosis and start the most effective therapy in case an ultrasonographic research should not result diriment. PMID- 12741347 TI - [Multicentric study for evaluation of levofloxacin in the treatment of complicated urinary tract infections]. AB - OBJECTIVES: The objective of this open-label, single-arm, multicenter study was to assess the efficacy and safety of levofloxacin in the treatment of complicated urinary tract infections. MATERIALS AND METHODS: A total of 72 outpatients with symptoms/signs of urinary tract infection and underlying anatomic or functional abnormality of the urinary tract was enrolled to receive levofloxacin 250 mg orally once daily for 10 days. Return visits were scheduled at 3-5 days (early follow-up visit) and 4-6 weeks after completion of therapy (long-term follow-up). At each visit symptoms and signs were assessed, possible adverse events were recorded and a urine specimen was obtained for culture. RESULTS: At 3-5 days post therapy clinical success rate was 94% and the overall microbiological eradication rate of pathogens was 94%. Microbiological eradication rate with respect to patients was 86%. Treatment failed in 4 patients and 2 patients had superinfections with pathogens still susceptible to levofloxacin. At long-term follow-up visit 3 reinfections were observed in patients previously cured at early follow-up. Drug-related adverse events were reported by 2 patients. CONCLUSION: These study results indicate that levofloxacin 250 mg orally once daily for 10 days is an effective and well tolerated treatment of complicated urinary tract infections. PMID- 12741348 TI - [Early use of menotropin in the treatment of varicocele]. AB - INTRODUCTION AND OBJECTIVES: Varicocele is the most common disease in infertile men and varicocelectomy remains the main treatment. During the last years a lot of drugs have been proposed in association with surgery. The aim of this study is verify the effects of the association between menotropin and subinguinal varicocelectomy on sperm parameters and pregnancy rate. METHODS: 60 patients with left varicocele were randomized in three groups of 20 patients: patients of group A were treated with menotropin since diagnosis for three months; patients of group B were treated with menotropin but started the treatment three months after surgical treatment; patients of group C were treated only with varicocelectomy. We have tested sperm parameters before and after three, six and twelve months from surgical treatment. After six and twelve months from varicocelectomy we have verified the pregnancy rates in the three groups. RESULTS: After three months from surgical treatment, sperm parameters were significantly improved only in the group A (p < 0.05). After six months we have recorded a similar improvement even in the group B. After twelve months we have confirmed a significant improvement in groups A and B; the values were significantly higher than in the group C (p < 0.05). After six months the pregnancy rate was 42%, 25% and 22.2% respectively in the three groups. After twelve months we have recorded an important improvement of pregnancy rate in groups A and B (47% and 45% respectively) and a little improvement (27.7%) in group C. CONCLUSIONS: Our data show a significant improvement of sperm parameters in the two groups treated with menotropin respect to the group treated with only varicocelectomy. It could be responsible for a similar improvement of the pregnancy rate. The association between varicocelectomy and early use of menotropin seem to improve testis functional rehabilitation. PMID- 12741349 TI - Unusual surgical approach to the treatment of renal cell carcinoma extending into the right atrium through the inferior vena cava. AB - We report an unusual surgical management of renal cell carcinoma with inferior vena cava and right atrial thrombosis. Successful management is dependent upon preoperative assessment to determine the extent of the disease, tumor thrombi has not infiltrated the walls of the vena cava, and a well planned surgical resection. PMID- 12741350 TI - Phlebography: why it is important to study radiological imaging of spermatic veins. AB - OBJECTIVES: We delineate the role of phlebography which is a standard procedure during antegrade and retrograde sclerotherapy of idiopathic varicoceles. MATERIAL AND METHODS: Based on our experience with antegrade sclerotherapy in 5,254 adults and adolescents we present typical phlebographies of the internal spermatic vein and radiological images after irregular cannulation of different arteries and veins on the scrotal floor. The sequalae of misinjections and paravascular application of the sclerosing agent polidocanol are discussed. A radiological classification of venous drainage roots of the pampiniform plexus is introduced on the basis of 236 consecutive antegrade phlebographies. The findings are compared with the results of retrograde phlebography previously given in literature. RESULTS: Phlebography helps to ascertain that sclerotherapy is performed in an anatomically appropriate manner. Thus, misinjections and paravascular applications of polidocanol can be ruled out. Antegrade phlebography enables to visualize a high rate of side veins and collaterals of the internal spermatic vein and thus seems to be equivalent to the retrograde imaging in tracing the additional drainage roots of the pampiniform plexus. These additional roots are considered to be the source of persisting varicoceles after surgical treatment. CONCLUSIONS: Phlebography is needed to achieve good results after sclerotherapy of varicoceles and to minimize the toxic side effects of polidocanol. Finally it is a prove that the treatment has been performed accurately for forensic reasons. PMID- 12741351 TI - Transrectal ultrasound imaging and prostate cancer. AB - Prostate cancer is one of the most important causes of death from cancer in men. Ultrasound imaging is frequently used in the diagnosis of prostate cancer. This paper presents an overview of currently available ultrasound imaging techniques. The underlying principles and methods are discussed briefly and examples of the clinical application are presented. Techniques such as gray-scale imaging, Doppler imaging and contrast enhanced imaging techniques using power Doppler, color Doppler, harmonic imaging and intermittent imaging are discussed. PMID- 12741352 TI - [Role of viral-herpetic infections in the etiology of atherosclerosis: clinical, virological and immunological evidence]. AB - The article describes the clinical, virological and immunological data confirming the etiological role of herpes virus in the initiation of atherosclerosis. 226 patients with atherosclerosis of the predominantly coronary localization were examined; hypertension and stenocardia were found in a part of them, while myocardial infarction was diagnosed in 22% of the patients. The control group consisted of patients with other diseases related with infections (bronchial asthma, rheumatism etc.) as well as of healthy persons. A total of 558 patients were examined and it was established that there is a reliable relation between atherosclerosis and the infection of patients with, mainly, herpes virus. The correlation was of the seasonal nature, it was linked to the specific features of an infection process and it was confirmed by the condition of the cholesterol supply and by immunodeficiency in patients. The infectious nature of atherosclerosis demands further research for the sake of finding proof of the etiological role of viruses and bacteria and for the sake of working out the means of prophylaxis and treatment of atherosclerosis aimed at removing the infectious etiological factor. PMID- 12741353 TI - [Influenza viruses and atherosclerosis: the role of atherosclerotic plaques in prolonging the persistent form of influenza infection]. AB - It was established that viral particles, like low-density lipoproteins (LDLP), when subjected to some modification changes, lost their ability to be internalized by tissue somatic cells and acquired tropism to macrophage cells. The data, obtained by us by using the polymerase chain reaction (PCR) method, made it possible to assert that atherosclerotic plaques, isolated from vessels of patients with ischemic heart disease (IHD) who underwent coronary bypass, contained RNA of the A(HINI) and AH3N3) influenza viruses. Whereas, the vessel portions, undamaged by atherosclerosis, did not contain any genetic substances of influenza viruses. It was for the first time that an experimentally supported understanding was expressed on that the atherosclerotic plaques serve as a "reservoir" for influenza viruses. It is also suggested that the mentioned plaques can be the carriers of influenza viruses for a long time, thus, prolonging the persistent form of influenza infection in the human body. PMID- 12741354 TI - [Specific features of the structure of sleep and personality under the conditions of chronic emotional stress and methods of improving the adaptive abilities of man]. AB - The results of a complex psychophysiological study of human sleep in the chronic stress condition are described. 20 patients with complaints of night sleep disturbances in the chronic stress condition and 10 healthy controls were examined by polysomnography. Profound changes of the quantitative and qualitative sleep parameters, mainly, related with the slow sleep phase, sleep delta characteristics and adaptive wakefulness mechanisms. The study demonstrated the effectiveness of improving the human adaptive parameters in the chronic stress conditions by using the drug and drugless therapy methods. PMID- 12741355 TI - [Chronopharmacological approaches to the therapeutic prophylaxis of chronic cardiac insufficiency in type I diabetes mellitus]. AB - The efficiency of the developed scheme for a drug prophylaxis of chronic cardiac insufficiency (CCI) in patients with diabetes mellitus I (DM I) of autoimmune genesis (subtype B) was evaluated; such scheme comprised the inhibitors of angiotensin converting enzyme (ACE) of different chemical structures and a biological response modifier (BRM)--glutoxime, which were prescribed with regard for a changed biological rhythm of hemodynamic as well as of metabolic and immune profiles of patients. The following parameters were examined: cardiohaemodynamics, the content of cortisol, insulin, ACTH, TTH and thyroxin in blood, of transport proteins and of the concentration of IL-1 and TNF-alpha in blood. It was established that the use of ramipril and fosinopril combined with the glutoxime BRM within the scheme of drug prophylaxis of CCI concomitant with DM I was accompanied by an effective removal of desynchronosis of the immune endocrine system in the discussed category of patients. PMID- 12741356 TI - [Generation of nitric oxide by peripheral blood leukocytes and platelets in thoracic and abdominal wounds]. AB - Generation of nitric oxide (NO) by human blood leukocytes and platelets was studied in 23 healthy persons and in 22 patients with thoracic and abdominal wounds. The sum of points ranged, in victims, from 12 to 19 according to the APACHE II severity-degree condition. Leukocytes and blood platelets were isolated from blood by differential centrifuging. The leukocyte suspension was diluted by an absolute medium (RPMI 1640, embryonic calf serum, L-glutamine, antibiotics) up to 5 x 10(6) cells in 0.5 ml; while the blood platelet suspension was diluted to 05 x 10(8) in 0.5 ml, and they were cultivated for 15 at 37 degrees C. The concentration of NO nitrite metabolite was determined by Griss reaction. The NO content in leukocyte supernatant of healthy persons amounted to 0.38 +/- 0.03 nmol/0.5 x 10(6) leukocytes; while in patients this figure essentially increased- up to 1.06 +/- 0.08 nmol/5 x 10(6) leukocytes, (p < 0.001). The NO content in blood-platelet supernatant amounted, in the normal condition, to 1.07 +/- 0.07 nmol/0.5 x 10(8) blood platelets; in the victims it was 1.52 +/- 0.11 nmol/L blood platelets, (p < 0.01). The relative production of NO by leukocytes was found to be, in healthy persons, 0.75 +/- 0.06 mumol/L, (p < 0.001), in patients- 3.05 +/- 0.22 mumol/L, (p < 0.01). The absolute NO concentration was determined with regard for an actual content of leukocytes and blood platelets in each healthy person and in each patient with thoracic and abdominal wounds. The absolute production of NO by leukocytes was, in healthy persons, 0.47 +/- 0.05 mumol/L, in patients--2.29 +/- 0.23 mumol/L, (p < 0.001). The absolute generation of NO by blood platelets was, in norm, 7.70 +/- 0.55 mumol/L, and in patients- 11.45 +/- 0.84 mumol/L, (p < 0.0011). Consequently, the absolute production of NO by blood platelets is by 16 times higher than the absolute production of NO by leukocytes in healthy people. Thus, stress enhances the NO generation by both leukocytes and blood platelets. PMID- 12741357 TI - [Endogenous biological retranslation in current clinical and biological studies]. AB - The paper generalizes the results of research implemented within the framework of the methodological concept of the Endogenous Biological Retranslation (EBR) for the purpose of finding new data about the immune pathogenesis of multiple sclerosis (MS), whose mechanisms are related with violations of the EBR processes in the body. A possibility is considered to use the priming of interferon (INF) production, as an EBR variant, for evaluating the functioning of the INF system. The thus obtained materials prove convincingly that the experimental approaches, based on priming the blood cells to the IFN production, can be clinically used in the diagnosis and therapy of MS. A new effective combined IFN-therapy scheme for the disease in question was demonstrated. A set of criteria, which makes it possible to prescribe the INF preparations for MS patients with regard for an evaluated response degree of patients' blood cells to them and with regard for a prognosticated efficacy of an assumed treatment is offered. Possibilities of an individualized comprehensive therapy for MS by using the offered experimental approaches are discussed. PMID- 12741359 TI - [Bioterrorism as a threat to safety. An evaluation of readiness degree of medical institutions to resist bioterrorism]. PMID- 12741358 TI - [Chronic viral hepatitis]. PMID- 12741360 TI - [Political factors and the Russian health care system]. PMID- 12741362 TI - [Academician of the USSR Academy of Medical Sciences V. V. Parin (on the occasion of the centennial of his birth)]. PMID- 12741361 TI - [Pathogenetic mechanisms of chronic urticaria]. PMID- 12741363 TI - Results of the first phase 3 trial of an AIDS vaccine. PMID- 12741364 TI - AIDS drug in halted trial less effective as monotherapy. PMID- 12741365 TI - Government guidelines for HIV/AIDS palliative care. PMID- 12741366 TI - Do we have a shared future, or not? PMID- 12741367 TI - Resistance studies from the 10th Conference on Retroviruses and Opportunistic Infections. PMID- 12741368 TI - Long-term complications of nucleoside reverse transcriptase inhibitor therapy. AB - HAART has resulted in dramatic declines in morbidity and mortality among patients infected with HIV. Increased experience with HAART has led to the detection of drug related toxicities that may compromise adherence and necessitate discontinuation of treatment and alteration of otherwise effective regimens. This article considers the major long-term complications associated with nucleoside reverse transcriptase inhibitor (NRTI) use--hyperlactatemia and lactic acidosis/hepatic steatosis, other hepatotoxicities, pancreatitis, lipodystrophy, lipoatrophy, neuropathy, and hematologic toxicities. Mechanisms by which NRTIs may produce these effects are discussed, as are differential effects of agents in this class and management options. PMID- 12741369 TI - Significant decrease in hemoglobin and hematocrit levels in a virologically controlled HIV-infected patient. Pure red cell aplasia (PRCA) caused by human parvovirus B19. PMID- 12741371 TI - Genetic privacy: the potential for genetic discrimination in insurance. PMID- 12741370 TI - Plasma homovanillic acid: a significant association with alcoholism is independent of a functional polymorphism of the human catechol-O methyltransferase gene. AB - The central dopamine system seems to influence addictive disorders. Plasma homovanillic acid (HVA) is an indicator of central dopaminergic activity. In this study the hypothesis that plasma HVA is associated with alcoholism or with delirium tremens (DT) during alcohol withdrawal was tested. A functional genetic polymorphism of the enzyme catechol-O-methyltransferase (COMT) that participates in converting dopamine into its final metabolite HVA was investigated for an association with alcoholism or DT during alcohol withdrawal. In addition, a relation between the functional polymorphism of COMT and plasma HVA concentrations was studied. Plasma HVA concentrations and COMT genotypes were determined in 142 German alcoholics and 101 German healthy controls. Alcoholic patients were examined after a minimum of 3 weeks after cessation of drinking. Mean plasma HVA concentrations were significantly lower in alcoholic patients compared to healthy controls. A group of alcoholics with a history of DT during alcohol withdrawal (n=62) did not differ significantly in plasma HVA concentrations from alcoholics with a history of only mild withdrawal symptoms (n=67). The functional polymorphism of the human COMT gene was neither significantly associated with the diagnosis of alcoholism or DT during alcohol withdrawal nor with plasma HVA concentrations. PMID- 12741372 TI - Wrongful life, wrongful birth, wrongful death, and the right to refuse treatment: can reasonable jurisdictions recognize all but one? PMID- 12741373 TI - Baby contracts. PMID- 12741375 TI - Animal experimentation and the First Amendment. PMID- 12741374 TI - Crossing the line: the political and moral battle over late-term abortion. PMID- 12741376 TI - Unwarranted government disclosures: reflections on privacy rights, HIV and ad hoc balancing. PMID- 12741377 TI - The allure and peril of genetics exceptionalism: do we need special genetics legislation? PMID- 12741378 TI - Genophobia: what is wrong with genetic discrimination? PMID- 12741379 TI - The international human right to health. PMID- 12741380 TI - [Breast cancer screening by mammography: evaluation by Gotzsche and Olsen meta analysis]. PMID- 12741381 TI - For aerospace programs, the crises continue. PMID- 12741382 TI - Square pegs in round holes: the dilemma of conjoined twins and individual rights. AB - The judgment in the English Court of Appeal case of Re A (Conjoined Twins: Surgical Separation) highlights forcefully the highly individualistic and abstract assumptions that commonly shape the deployment of rights discourse in liberal legal adjudication. Forced by the all-or-nothing nature of this discourse into a dilemma between perceiving of the twins as separate right-bearers or perceiving of the stronger twin, Jodie, as the singular right-bearer and of Mary, her weaker sibling, as a non-legal entity, the court chose the former option. Perceiving of the twins as distinct and equal legal persons forced the court to employ a balancing of incommensurate interests, implicitly accepting a utilitarian analysis within the strongly deontological confines of law and medicine. The implications of this turn towards utilitarianism are significant. Within the confines of this article, it will be argued, however, that these implications are avoidable if the law concedes a more flexible approach to the dominant notion of the distinct and autonomous right-bearer. PMID- 12741383 TI - Population-based genetic studies: informed consent and confidentiality. PMID- 12741384 TI - Bio-ethics and physician liability: the liability effects of developing pain management standards. PMID- 12741385 TI - Ethics and e-medicine. PMID- 12741386 TI - DNA analysis and the Freedom of Information Act: information or invasion? AB - This note explores the possibility of release of an individual's DNA analysis to any person who requests it through the Freedom of Information Act (FOIA), after an individual's post-aircraft accident DNA profile has been developed by the Federal Aviation Administration's (FAA) Civil Aerospace Medical Institute (CAMI). It analyzes whether the request would fall under the FOIA's 552(b)(6) exemption, which weighs a person's privacy interest against any public interest in such information, or if the release would constitute a "clearly unwarranted invasion of personal privacy." PMID- 12741387 TI - International conflicts over patenting human DNA sequences in the United States and the European Union: an argument for compulsory licensing and a fair-use exemption. AB - The thought of a large biotech company holding an exclusive right to research and manipulate human genetic material provokes many reactions--from moral revulsion to enthusiasm about the possibilities for therapeutic advancement. While most agree that such a right must exist, debate continues over the appropriate extent of its entitlements and preclusive effects. In this Article, Professor Donna Gitter addresses this multidimensional problem of patents on human deoxyribonucleic acid (DNA) sequences in the United States and the European Union. Professor Gitter chronicles not only the development of the law in this area, but also the array of policy and moral arguments that proponents and detractors of such patents raise. She emphasizes the specific issue of patents on DNA sequences whose function has not fully been identified, and the chilling effect these patents may have on beneficial research. From this discussion emerges a troubling realization: While the legal framework governing "life patents" may be similar in the United States and the European Union, the public perceptions and attitudes toward them are not. Professor Gitter thus proposes a dual reform: a compulsory licensing regime requiring holders of DNA sequence patents to license them to commercial researchers, in return for a royalty keyed to the financial success of the product that the licensee develops; and an experimental-use exemption from this regime for government and nonprofit researchers. PMID- 12741389 TI - Court-authorised caesarean sections--the end of a trend? PMID- 12741388 TI - Adsorption processes in spacecraft environmental control and life support systems. AB - The environmental control and life support system on a spacecraft maintains a safe and comfortable environment in which the crew can live and work by supplying oxygen and water and by removing carbon dioxide, water vapor, and trace contaminants from cabin air. Although open-loop systems have been used successfully in the past for short-duration missions, the economics of current and future long-duration missions in space will make nearly complete recycling of air and water imperative. A variety of operations will be necessary to achieve the goal of nearly complete recycling. These include separation and reduction of carbon dioxide, removal of trace gas-phase contaminants, recovery and purification of humidity condensate, purification and polishing of wastewater streams, and others. Several of these can be performed totally or in part by adsorption processes. These processes are good candidates to perform separations and purifications in space due to their gravity independence, high reliability, relative high energy efficiency, design flexibility, technological maturity, and regenerative nature. For these reasons, adsorption has historically played a key role in life support on U.S. and Russian piloted spacecraft. Among the life support applications that can be achieved through use of adsorption technology are removal of trace contaminants and carbon dioxide from cabin air and recovery of potable water from waste streams. In each of these cases adsorption technology has been selected for use onboard the International Space Station. The requirements, science, and hardware for these applications are discussed. Human space exploration may eventually lead to construction of planetary habitats. These habitats may provide additional opportunities for use of adsorption processes, such as control of greenhouse gas composition, and may have different resources available to them, such as gases present in the planetary atmosphere. Separation and purification processes based on adsorption can be expected to continue to fulfill environmental control and life support needs on future missions. PMID- 12741390 TI - Back to the moon: the lure of lunar exploration. PMID- 12741391 TI - Consent or property? Dealing with the body and its parts in the shadow of Bristol and Alder Hey. AB - This article first considers the tenuous base on which the law of property in the body is founded, and then discusses the practical results of this in the light of the recent furore surrounding events at Bristol and Alder Hey. The authors suggest that neither the consent-based model followed by the official inquiries into these events nor a possible policy based on a full-blown property model adequately cover the private rights of an individual's next of kin or the right of the public to an efficient and reliable pathological service within the NHS. Rather, they propose that a combined model in which a 'cascade of possession' for the recognition of various property interests is initiated by assent on the part of the next of kin and terminates in full possession of the body vested in the execution for the purposes of its disposal. The authors recommend further that any reform of the law should apply property rights to body parts taken from both the living and the dead. PMID- 12741392 TI - Cellular fluid mechanics. AB - The coupling of fluid dynamics and biology at the level of the cell is an intensive area of investigation because of its critical role in normal physiology and disease. Microcirculatory flow has been a focus for years, owing to the complexity of cell-cell or cell-glycocalyx interactions. Noncirculating cells, particularly those that comprise the walls of the circulatory system, experience and respond biologically to fluid dynamic stresses. In this article, we review the more recent studies of circulating cells, with an emphasis on the role of the glycocalyx on red-cell motion in small capillaries and on the deformation of leukocytes passing through the microcirculation. We also discuss flows in the vicinity of noncirculating cells, the influence of fluid dynamic shear stress on cell biology, and diffusion in the lipid bi-layer, all in the context of the important fluid-dynamic phenomena. PMID- 12741394 TI - An unusual rash. Cholesterol embolism. PMID- 12741396 TI - Professional misconduct. PMID- 12741395 TI - Measuring surface-area-to-volume ratios in soft porous materials using laser polarized xenon interphase exchange nuclear magnetic resonance. AB - We demonstrate a minimally invasive nuclear magnetic resonance (NMR) technique that enables determination of the surface-area-to-volume ratio (S/V) of soft porous materials from measurements of the diffusive exchange of laser-polarized 129Xe between gas in the pore space and 129Xe dissolved in the solid phase. We apply this NMR technique to porous polymer samples and find approximate agreement with destructive stereological measurements of S/V obtained with optical confocal microscopy. Potential applications of laser-polarized xenon interphase exchange NMR include measurements of in vivo lung function in humans and characterization of gas chromatography columns. PMID- 12741393 TI - FAK induces expression of Prx1 to promote tenascin-C-dependent fibroblast migration. AB - Fibroblast migration depends, in part, on activation of FAK and cellular interactions with tenascin-C (TN-C). Consistent with the idea that FAK regulates TN-C, migration-defective FAK-null cells expressed reduced levels of TN-C. Furthermore, expression of FAK in FAK-null fibroblasts induced TN-C, whereas inhibition of FAK activity in FAK-wild-type cells had the opposite effect. Paired related homeobox 1 (Prx1) encodes a homeobox transcription factor that induces TN C by interacting with a binding site within the TN-C promoter, and it also promotes fibroblast migration. Therefore, we hypothesized that FAK regulates TN-C by controlling the DNA-binding activity of Prx1 and/or by inducing Prx1 expression. Prx1-homeodomain binding site complex formation observed with FAK wild-type fibroblasts failed to occur in FAK-null fibroblasts, yet expression of Prx1 in these cells induced TN-C promoter activity. Thus, FAK is not essential for Prx1 DNA-binding activity. However, activated FAK was essential for Prx1 expression. Functionally, Prx1 expression in FAK-null fibroblasts restored their ability to migrate toward fibronectin, in a manner that depends on TN-C. These results appear to be relevant in vivo because Prx1 and TN-C expression levels were reduced in FAK-null embryos. This paper suggests a model whereby FAK induces Prx1, and subsequently the formation of a TN-C-enriched ECM that contributes to fibroblast migration. PMID- 12741397 TI - Analysis and experiments with an elephant's trunk robot. AB - The area of tentacle and trunk type biological manipulation is not new, but there has been little progress in the development and application of a physical device to simulate these types of manipulation. Our research in this area is based on using an 'elephant trunk' robot. In this paper, we review the construction of the robot and how it compares to biological manipulators. We then apply our previously designed kinematic model to describe the kinematics of the robot. We finish by providing some examples of motion planning and intelligent manipulation using the robot. PMID- 12741398 TI - Over 2.2 million low-income California adults are food insecure; 658,000 suffer hunger. PMID- 12741399 TI - Diabetes in California: nearly 1.5 million diagnosed and 2 million more at risk. PMID- 12741400 TI - Small, inexpensive device could revolutionize HIV testing. PMID- 12741401 TI - [Methicillin-resistant Staphylococcus aureus (MRSA) infections spread in USA]. PMID- 12741402 TI - Fragmented bodies, ideal privilege, and commodification in science and medicine. PMID- 12741403 TI - Separating conjoined twins: legal reverberations of Jodie and Mary's predicament. PMID- 12741404 TI - Giving until it hurts: prisoners are not the answer to the national organ shortage. AB - This Note argues that prisoners, whether executed or living, should not become organ donors. The introduction acknowledges the shortage of transplantable organs in the United States and the steps that have been taken to ameliorate the crisis. Part I discusses the procurement of organs from executed prisoners, beginning with a brief examination of China, a country where this type of procurement is routinely practiced. Part I also examines organ procurement legislation pertaining to executed prisoners. Finally, Part I asserts the reasons that prisoners should not become donors, including the dead donor rule, the ban against physicians as executioners, the Oath of Hippocrates, the risk of transmissible diseases, and the negative perception that would result if organ procurement was tied to executions. Part II of this Note discusses prisoners donating their organs in return for mitigated sentences. Part II then argues that this practice should not be adopted because of the lack of informed consent and voluntary choice. Finally, Part III of this Note introduces potential solutions to the possibility of maintaining a voluntary system, moving to a presumed consent system, and using financial inducements to create a larger supply of transplantable organs. PMID- 12741405 TI - [Using low osmolar gadolinium for angiographic imaging and in interventional radiologic procedures in patients with renal failure]. PMID- 12741406 TI - The epitome of a nurse. PMID- 12741407 TI - A 67-year-old female with a 4-year history of type 2 diabetes, treated with metformin, presented with unstable angina. PMID- 12741408 TI - Is the PRIME (Primary Response In Medical Emergencies) scheme acceptable to rural general practitioners in New Zealand? AB - AIM: To ascertain the level of acceptance of the PRIME (Primary Response In Medical Emergencies) scheme by rural general practitioners (GPs) in New Zealand. METHODS: A nationwide, anonymous, postal/email questionnaire was sent to 536 rural/semi-rural GPs, inquiring as to their involvement in and opinions of emergency care, and the acceptability of the PRIME scheme. RESULTS: The overall response rate was 42%. PRIME training courses and PRIME equipment were regarded as excellent. However, concerns were raised by both PRIME and non-PRIME groups regarding the quality of triaging information given during emergencies and levels of remuneration for call-outs (especially medical call-outs). Additional concerns included lack of flexibility with the PRIME contract in some areas. Some GPs were also concerned that their involvement was less about providing a higher skill level in resuscitation than about filling the gaps in the already-stretched rural ambulance services, which was not the intention of the PRIME scheme. CONCLUSIONS: The inclusion of rural GPs in emergency care teams needs to be recognised and adequately remunerated, and these issues should be reflected in the ongoing development of pre-hospital emergency service contracts. PMID- 12741409 TI - [Lumbar meningeal enhancement in contrast MRI after operation of the posterior skull base: a normal occurrence in children?]. PMID- 12741410 TI - Independent Institutional Review Boards. PMID- 12741411 TI - Investigator, IRB and institutional financial conflicts in interest in human subject research: past, present and future. PMID- 12741412 TI - Remaining faithful to the promises given: maintaining standards in changing times. PMID- 12741413 TI - The line between clinical innovation and human experimentation. PMID- 12741414 TI - Using scientific advances to conceive the "perfect" donor: the Pandora's box of creating child donors for the purpose of saving ailing family members. PMID- 12741415 TI - Does vitamin E supplementation prevent cardiovascular events? AB - In recent years, vitamin E has been investigated as a cardioprotective agent. Experimental studies have identified potential mechanisms by which vitamin E may inhibit the development of atherosclerosis, and observational studies of individuals without coronary disease suggest that vitamin E intake may prevent future cardiovascular events. Secondary prevention trials to date have demonstrated little benefit from vitamin E supplementation. It remains possible, however, that supplementation may be useful among certain high-risk groups, including those with nutritional deficiencies. Limited data from completed primary prevention trials also indicate minimal cardioprotection from vitamin E, but large-scale trials now in progress may yet show benefit. Results from ongoing trials will contribute powerfully to the totality of evidence on which to formulate both appropriate clinical recommendations for individual patients and a rational public health policy for the population as a whole. At this time, there is insufficient evidence for issuing a public health recommendation to use vitamin E supplements to prevent cardiovascular disease (CVD). Rather, increased intake of fruits, vegetables, and other antioxidant-rich foods should be promoted as part of a healthy diet because they provide nutritional benefits beyond any potential antioxidant effect. Moreover, even if found to reduce CVD risk, vitamin supplement use should be considered an adjunct, not an alternative, to established cardioprotective measures, such as smoking abstention, avoidance of obesity, adequate physical activity, and control of high blood pressure and hyperlipidemia. PMID- 12741416 TI - The missing R. PMID- 12741417 TI - [Quality control of culture media for Mycobacterium]. PMID- 12741418 TI - Cross-sectional area of a bone-patellar tendon-bone graft for anterior cruciate ligament reconstruction. AB - The cross-sectional area of the 10-mm wide patellar tendon graft was measured in 50 consecutive patients (31 males and 19 females) who underwent isolated anterior cruciate ligament (ACL) reconstruction and the relationship between the graft size and various factors such as physical characteristics was assessed. The effect of cross-sectional area of the implanted graft on postoperative stability of the reconstructed knee also was examined. Mean patient age at surgery was 22.3 years (range: 14-40 years). The cross-sectional area was measured using an instrumented area micrometer intraoperatively, and correlations between the measured value and various factors such as age, gender, height, body weight, and bony geometry were examined. Follow-up was performed 24 months postoperatively. The average cross-sectional graft area was 33.4 mm2. The measured cross-sectional area was larger in male patients and correlated with physical characteristics such as height, body weight, and femoral condyle width. No significant correlation between the size of the graft and postoperative stability was observed. PMID- 12741419 TI - Meniscal repair with the Rapid Loc device: a cadaveric study. AB - The devices available for all-inside meniscal repair were evaluated using a cadaver model. Rapid Loc devices (Mitek, Westwood, Mass) were inserted arthroscopically into 8 fresh-frozen cadaveric knees (age range: 66-78 years) in the posterior regions of the menisci at 5- to 7-mm intervals. Forty-eight devices were placed (3 in each meniscus), and the knees were subsequently dissected to determine the location of the inserted devices. Devices were correctly inserted in 40 (833%) of 48 knees. No problems were noted with loss of fixation on the periphery of the meniscus. Entrapment of the popliteus tendon (3 of 8) and superficial medial collateral ligament (2 of 16) were the only major problems with the device. PMID- 12741420 TI - The microfracture technique in the treatment of full-thickness chondral lesions of the knee in National Football League players. AB - Between 1986 and 1997, 25 active National Football League (NFL) players underwent microfracture to treat full-thickness chondral lesions. Average follow-up was 4.5 years (range: 2-13 years). Preoperative and postoperative data were rated according to symptoms (4 = severe, 1 = none), function (5 = unable to perform, 1 = no limitations), and activity level (10 = full activity, 1 = unable to perform). Nineteen (76%) players returned to football the season following microfracture. Six players retired for various reasons. At follow-up, pain, swelling, running, cutting, and squatting improved. Activities of daily living, strenuous work, and strenuous sport levels also improved. Those who returned to play averaged 4.6 seasons of participation (range: 1-13 seasons) and 56 games (range: 2-183 games) after microfracture. Nine (36%) players continue active participation in the NFL. Results of microfracture to resurface full-thickness chondral lesions in high-demand NFL players are encouraging. Microfracture is safe, effective, and appears to improve symptoms, function, and activity levels in NFL players. PMID- 12741421 TI - Effect of functional bracing on subsequent knee injury in ACL-deficient professional skiers. AB - The effect of functional bracing on subsequent knee injury in anterior cruciate ligament (ACL) deficient professional skiers was evaluated. A cohort of 180 ACL deficient skiers was identified from a knee screening of 9410 professional skiers from 1991-1997. An ACL-deficient knee was defined by an abnormal examination (Lachman or pivot-shift) and a > or = 5-mm KT-1000 manual maximum difference. The dependent variable was subsequent knee injury, which occurred in 12 knees. A significantly higher proportion of injuries occurred in nonbraced skiers compared with braced skiers (P = .005). The risk ratio for subsequent knee injury comparing nonbraced with braced skiers was 6.4 (13% and 2%, respectively). Univariate analysis revealed no significant effects of the other covariates. Logistic regression identified bracing status (P < .01; odds ratio = 8) and KT 1000 manual maximum difference (P = .02; odds ratio = 1.3) as significant multivariate risk factors for subsequent knee injury, controlling for covariates. PMID- 12741422 TI - Cytokine levels after transfusion of washed wound drainage in total knee arthroplasty: a randomized trial comparing autologous blood and washed wound drainage. AB - Forty patients undergoing total knee arthroplasty (TKA) were prospectively randomized to autologous blood transfusion or retransfusion of washed wound drainage. The wound drainage was washed using a dynamic disk separation chamber. Interleukin (IL)-1 beta, IL-6, IL-8, and tumor necrosis factor (TNF) alpha blood levels were measured prior to epidural anesthesia and at 0, 30, 60, and 120 minutes after initiation of the transfusion. Washed wound drainage contained significantly higher amounts of IL-6 (P < .05) and IL-8 (P < .05) than predonated autologous blood. No significant difference was found for IL-1 beta (P = .28) and TNF-alpha (P = .14). The patients' IL-6 and IL-8 blood levels increased in both groups within 2 hours after the transfusion. This increase was correlated to the time interval between surgery and transfusion (P < .05) and was not significantly different between both groups. No correlation was found between the increase in patients' cytokine levels and the amount of washed wound drainage transfused. According to these results, the increase in IL-6 and IL-8 after transfusion is related to the surgical trauma response and not the transfusion protocol. PMID- 12741423 TI - Accuracy evaluation of a shape-based registration method for a computer navigation system for total knee arthroplasty. AB - This study evaluated the effect of computed tomography (CT) slice thickness, reconstruction pitch, intraoperative data sampling area, and data sampling volume on the accuracy of registration and determined a clinically acceptable trade-off between accuracy and surgical invasiveness. One cadaveric femur and one cadaveric tibia were used. Computed tomography of the femur and tibia were obtained using a helical scanner. Three sets of slice thickness and slice pitch were chosen for data acquisition, and two additional sets of reconstructed data were made. Bone contours were extracted by removing surrounding substrate. Surface models of bones were made from the resulting data. Registration of surface models to real objects was performed by measuring the position of various surface points on various areas of each object using an OPTOTRAK pen-probe (Northern Digital Inc, Ontario, Canada). The following trade-off is proposed as clinically optimal: perform CT with 3-mm slice thickness and 1-mm reconstruction pitch, and sample a periarticular area of 30 sampling points. The accuracy of registration in terms of position and angle was 0.8 mm and 0.6 degrees of bias with 0.2 mm and 0.3 degrees of root-mean-square in the femur, and 0.5 mm and 0.4 degrees of bias with 0.2 mm and 0.3 degrees of root-mean-square in the tibia. PMID- 12741424 TI - Primary intra-articular liposarcoma of the knee. Case report. PMID- 12741425 TI - Treatment of periprosthetic fractures around a total knee arthroplasty. PMID- 12741426 TI - Extensor mechanism problems following total knee replacement. AB - Despite advances in TKR, a number of complications related to the extensor mechanism still exist. The vast majority of these are preventable with good surgical technique and attention to detail. Proper component positioning, adequate soft-tissue balancing and dissection, and gentle retraction are imperative. Proper resection and resurfacing of the patella, central tracking of the implant, maintenance of the patella vascularity, and use of an implant with a well designed patellofemoral joint help minimize complications of TKR. PMID- 12741427 TI - Flexion instability in primary total knee replacement. AB - Although the results of TKR are highly successful at long-term follow-up, failures occur. One of the more frequent causes of failure is instability. In distinction to instability in the medial-lateral plane, AP instability in flexion has been poorly described until recently. Although acquired ligamentous incompetence can occur, particularly with cruciate retaining prostheses, many cases of flexion instability result from an intraoperative failure to create symmetric balanced flexion and extension spaces. In primary TKR, use of a well designed posterior stabilized prosthesis and creation of symmetric balanced flexion and extension gaps should minimize the incidence of postoperative flexion instability. If flexion instability occurs, the role of nonoperative treatment is limited. In most cases, revision TKR using the same basic principles is required. When symmetric flexion and extension spaces cannot be produced intraoperatively in complex primary or revision surgery, use of a more constrained articulation, such as a constrained condylar prosthesis or hinged prosthesis, is required. PMID- 12741428 TI - High-dose hepatitis B vaccine in patients waiting for lung transplantation. AB - STUDY OBJECTIVE: To increase the response rate to hepatitis B vaccine in patients awaiting lung transplantation. DESIGN: Historically controlled, open-label study. SETTING: Lung transplant clinic at a university hospital. SUBJECTS: Twenty-seven consecutive individuals with end-stage pulmonary disease who were enrolled to accrue 15 subjects who would complete the vaccine series before transplantation; and 27 lung transplant recipients who were immunized with the conventional dose before the study and served as historical controls. INTERVENTION: Intramuscular injection of high-dose hepatitis B vaccine 40 microgram at 0, 1, and 6 months. MEASUREMENTS AND MAIN RESULTS: Hepatitis B surface antibody (anti-HBs) concentrations were measured 1-2 months after completing the high-dose series. Individuals with undetectable anti-HBs received additional vaccine to a maximum of six doses. The response rate to the series was compared with that in the control group. Seventeen individuals in the high-dose group and 14 controls met the study criterion of complete vaccine series before transplantation. The former had a much higher response rate than the latter (9 [53%] vs 1 [7%], p<0.01). Four of six patients who received additional doses of vaccine seroconverted. Two of them underwent transplantation shortly after completing the three-dose series. CONCLUSION: The high-dose hepatitis B vaccine series produced a protective immune response in lung transplant recipients; however, the response was suboptimal, and alternative immunization strategies should be studied. PMID- 12741429 TI - Estradiol in premenstrual asthma: a double-blind, randomized, placebo-controlled, crossover study. AB - STUDY OBJECTIVES: To characterize asthma symptoms and pulmonary function throughout two menstrual cycles, with and without exogenous estradiol administration, in women with premenstrual asthma, and to determine the effect of estradiol administration on asthma symptoms, pulmonary function, quality of life, and biomarkers of airway inflammation. DESIGN: Double-blind, randomized, placebo controlled, crossover study. SETTING: Respiratory clinic and clinical research center. SUBJECTS: Twelve women with documented premenstrual asthma (> or = 20% premenstrual worsening of asthma symptoms and/or of peak expiratory flow [PEF] during a 1-month screening phase). INTERVENTION: Each woman received either estradiol 2 mg or placebo orally between cycle days 23 and 28 (i.e., premenstrually, or before the onset of menses) in the first cycle and then crossed over to the other arm in the second cycle. Throughout both cycles, the women recorded daily morning and evening PEF readings and asthma symptoms. MEASUREMENTS AND MAIN RESULTS: Spirometry testing and measurement of serum estradiol and biomarkers of airway inflammation were performed on days 8 (follicular phase), 22 (luteal phase), and 28 (premenstrually) of both the estradiol and placebo cycles. During the two premenstrual visits, the Asthma Quality of Life Questionnaire was administered. No notable differences were observed between the estradiol and placebo cycles in daily PEF recordings or composite asthma symptoms scores. The area under the curve (AUC) for the composite asthma symptoms versus time profile was numerically, but not statistically, lower (denoting less severe symptoms) during the estradiol cycle than during the placebo cycle. Likewise, no significant difference in AUC values for morning PEF or evening PEF was found between the estradiol cycle and the placebo cycle. Despite differences (p<0.05) in day-28 estradiol concentrations for estradiol and placebo cycles, no significant differences were found in forced expiratory volume in 1 second, serum endothelin-1, serum and urine eosinophil protein X, urine leukotriene E4, or quality-of-life scores. CONCLUSION: Exogenously administered estradiol did not have a significant effect in women with premenstrual asthma whose asthma was classified predominantly as mild and under excellent control. As in the case of premenstrual syndrome, the placebo effect may be prominent in premenstrual asthma. Further trials, involving women with more severe asthma under poorer control, are warranted to discern underlying mechanisms for the worsening of asthma in relation to menstruation. PMID- 12741430 TI - Variability in polyene content and cellular toxicity among deoxycholate amphotericin B formulations. AB - STUDY OBJECTIVE: To evaluate the toxicity of amphotericin B deoxycholate formulations. DESIGN: In vitro experiment. SETTING: University research center. MATERIAL: Human mononuclear THP-1 cells. INTERVENTION: The human mononuclear cells were exposed in vitro for 2 hours to the following deoxycholate formulations of amphotericin B, in 2.5- and 5-micro/ml concentrations: Apothecon, Pharmacia, Sigma, Gensia, Pharma-Tek, and VHA. MEASUREMENTS AND MAIN RESULTS: Toxicity of the amphotericin B formulations were assessed by measuring interleukin (IL)-1beta expression in an in vitro model. Amphotericin B content was measured by enzyme-linked immunosorbent assay (ELISA), and amphotericin A and B contents were assessed by spectrophotometry. Endotoxin contamination was evaluated in all reagents. Expression of IL-1beta from Sigma, Pharmacia, and Pharma-Tek formulations was increased approximately 250%, 50%, and 25%, respectively, compared with amphotericin A. Amphotericin B content of Sigma, Pharmacia, Pharma-Tek, and Gensia formulations, as measured by ELISA, was increased approximately 450%, 200%, 200%, and 100%, respectively, compared with Apothecon. This variation could not be explained by differences in amphotericin A or B content as measured by spectrophotometry. CONCLUSION: Amphotericin B is obtained from a fermentation plant and manufactured as a pharmaceutical at different facilities. Both previous clinical observations and the current in vitro evaluation revealed significant differences among the formulations. Likely, other polyenes or pyrogenic toxins in differing amounts are in these formulations, thus explaining the variability in toxicity observed among the formulations. PMID- 12741433 TI - Enteral fluconazole population pharmacokinetics in patients in the surgical intensive care unit. AB - STUDY OBJECTIVE: To determine the population pharmacokinetic parameters of enterally administered fluconazole in patients in a surgical intensive care unit (SICU). DESIGN: Population pharmacokinetics component of a prospective, randomized clinical study. SETTING: The SICU at a university hospital. PATIENTS: One hundred ten patients with an expected length of stay in the SICU of 3 or more days and a need for intubation, in whom at least one fluconazole plasma concentration-time measurement was available. Intervention. Patients received fluconazole as an 800-mg loading dose and as a 200- or 400-mg (depending on renal function) daily maintenance dose. Fluconazole suspension was administered enterally followed by a 30-ml free water flush. MEASUREMENTS AND MAIN RESULTS: Plasma samples were collected, and population pharmacokinetic analysis was performed with NONMEM software; a one-compartment pharmacokinetic model was used. Fluconazole clearance was dependent on creatinine clearance, and volume of distribution was dependent on body weight and age. In patients with creatinine clearance values greater than 80 ml/minute, between 30 and 80 ml/minute, and less than 30 ml/minute, geometric mean (percentage coefficient of variation) fluconazole clearance was 14.39 ml/minute (21%), 10.53 ml/minute (28%), and 5.47 ml/minute (30%), respectively. The geometric mean (percentage coefficient of variation) volume of distribution in all patients was 1.27 L/kg (28%) and decreased with increasing age. CONCLUSIONS: Fluconazole clearance values in patients in the SICU who had normal renal function and in those with renal impairment were in agreement with previously reported data. Fluconazole volume of distribution was larger and half-life was longer in the SICU population than in healthy subjects. PMID- 12741431 TI - Aspirin dosage and thromboxane synthesis in patients with vascular disease. AB - STUDY OBJECTIVE: To determine whether urinary 11-dehydrothromboxane B2 (d-TXB2) is a marker of aspirin resistance and define the relationship between aspirin dosage and concentrations of this thromboxane metabolite. DESIGN: Randomized, crossover study. SETTING: Two outpatient clinical centers. PATIENTS: Forty-eight patients (mean age 70 yrs) with vascular disease (52% clinical coronary artery disease, 29% cerebrovascular disease, 46% atrial fibrillation). INTERVENTION: Levels of serum thromboxane B2 and d-TXB2 were measured after patients were treated initially with aspirin 325 mg/day for 4 weeks, then again after random assignment to receive aspirin 81, 325, or 1300 mg/day for 4 weeks, and then again after resumption of 325 mg/day for 4 weeks. MEASUREMENTS AND MAIN RESULTS: During treatment with aspirin 325 mg/day, the mean +/- SD serum thromboxane B2 level was 0.9 +/- 1.2 ng/ml and median (interquartile range) was 0.4 (0.2-0.9) ng/ml. Mean urinary d-TXB2 was 16 +/- 7.9 ng/mmol creatinine, with a median of 15 (9.9-23) ng/mmol creatinine with aspirin 325 mg/day. After 4 weeks of aspirin 81 mg/day, levels of serum thromboxane B2 (p<0.01) and urinary d-TXB2 (p=0.04) were both significantly higher compared with aspirin 325 mg/day; for urinary d-TXB2, the median increase was 3.0 ng/mmol creatinine. After 4 weeks of treatment with aspirin 1300 mg/day, levels of serum thromboxane B2 (p<0.01) and urinary d-TXB2 (p<0.01) were both significantly lower compared with aspirin 325 mg/day; the median decrease in urinary d-TXB2 was 4.4 ng/mmol creatinine. CONCLUSION: Different aspirin dosages significantly affect serum and urinary markers of thromboxane synthesis. PMID- 12741432 TI - A gentamicin pharmacokinetic population model and once-daily dosing algorithm for neonates. AB - STUDY OBJECTIVE: To develop a gentamicin pharmacokinetic population model and once-daily dosing algorithm for neonates younger than 10 days. DESIGN: Prospective, open-label study. SETTING: Neonatal intensive care unit. PATIENTS: One hundred thirty-nine neonates prescribed gentamicin. MEASUREMENTS AND MAIN RESULTS: Gentamicin peak and trough serum concentrations were collected from 139 neonates divided into three groups who were receiving one of the following intravenous 24-hour gentamicin regimens during the first 10 days of life, based on gestational age and birth weight (group 1, < 28 wks, 2.5 mg/kg; group 2, 28-34 wks, 3 mg/kg; and group 3, > 34 wks, 4 mg/kg). A structural model was developed in ADAPT II software using a MAP Bayesian approach. Final population parameter estimates were calculated using iterative two-stage analysis. The median (range) gestational age and birth weight, respectively, were 32 weeks (23-42 wks) and 1.92 kg (0.47-5.00 kg). The final one-compartmental linear model had a median (range) gentamicin total clearance, half-life, and volume of distribution of 0.0709 L/hour (0.0151-0.246 L/hr), 8.59 hours (4.88-16.9 hrs), and 0.262 L (0.0903-0.929 L), respectively. Total clearance increased as gestational age increased (p<0.001). Group 1 (10.2 hrs) had a significantly longer half-life than either group 2 (8.89 hrs, p<0.01) or group 3 (6.98 hrs, p<0.01). Total clearance was associated with gestational age and birth weight: clearance (L/hr) = (0.00504 + [0.00108 x gestational age]) x birth weight (coefficient of determination [r2] = 0.897), and volume of distribution was associated with birth weight (r2 = 0.700). The following dosing algorithm was designed to reach a therapeutic 24 hour area under the curve (87.5 mg/L x hr) in neonates during the first 10 days after birth: 24-hour gentamicin dose (mg) = (0.441 + [0.0945 x gestational age]) x birth weight. CONCLUSION: This dosing algorithm provides a new approach for determining initial gentamicin dosing regimens in neonates; however, clinical validation is required. PMID- 12741434 TI - Assessment of differences in antimicrobial effect determined with two in vitro pharmacodynamic models: impact of surface area to volume ratio. AB - STUDY OBJECTIVE: To measure the influence of different surface area:volume ratios (SA:Vs) on antibiotic penetration and subsequent antibacterial effect. DESIGN: In vitro laboratory experiment. SETTING: Two academic research laboratories. INTERVENTION: The two models with effective SA:Vs of 5.34 and 4.80 cm(-1) were evaluated by conducting a time-kill experiment with Pseudomonas aeruginosa ATCC 27853 and ceftazidime. MEASUREMENTS AND MAIN RESULTS: Ceftazidime was administered by constant infusion into the central compartment. Its penetration into the peripheral compartment and bacterial counts were determined over 24 hours, and antibacterial effect was quantified. Antibiotic penetration, calculated using central compartment and peripheral compartment area under the concentration-time curves, and effect, quantified as the relationship between the areas under growth and kill curves, differed between the models. Antibiotic penetration into the peripheral compartment was 53% greater over the first 4 hours of the experiment in the model with the larger SA:V. This was associated with antibacterial effects that were 64% and 38% greater in the 0-4-hour and 0-24 hour time periods, respectively. CONCLUSION: Differences in antibiotic penetration and effect observed between these models are likely explained by differences in SA:V PMID- 12741435 TI - Illicit methylphenidate use in an undergraduate student sample: prevalence and risk factors. AB - STUDY OBJECTIVES: To assess the prevalence of illicit methylphenidate use among undergraduate college students at a large university, and to identify alcohol and other drug use behaviors, as well as the negative consequences and risk factors, associated with illicit methylphenidate use. DESIGN: Internet survey. SETTING: Large public university. SUBJECTS: Thirty-five hundred randomly selected undergraduate students. MEASUREMENTS AND MAIN RESULTS: Of the 2250 students who completed the survey, 3% reported past-year illicit methylphenidate use. Illicit methylphenidate users were significantly more likely to use alcohol and drugs and report adverse alcohol- and drug-related consequences than prescription stimulant users or students who did not use stimulants. Undergraduate men and women were equally likely to report past-year illicit methylphenidate use. Weekly party behavior was significantly associated with past-year illicit methylphenidate use. CONCLUSION: Illicit use of prescription-only stimulants on college campuses is a potentially serious public health issue. More work is needed to promote understanding and awareness of this problem among clinicians and researchers. PMID- 12741436 TI - The effect of an antimicrobial restriction program on Pseudomonas aeruginosa resistance to beta-lactams in a large teaching hospital. AB - STUDY OBJECTIVES: To compare the use of beta-lactams and subsequent Pseudomonas aeruginosa sensitivity patterns before and after implementation of a clinical pharmacist-facilitated antimicrobial restriction program in August 1997. DESIGN: Retrospective consecutive data collection. SETTING: Large university-affiliated medical center. INTERVENTION: The study results are the accumulation of the daily intervention activities of the antimicrobial restriction program. Data on antimicrobial grams purchased/1,000 patient-days and susceptibility patterns were collected and analyzed retrospectively. MEASURES AND MAIN RESULTS: Annual grams of ceftazidime, piperacillin, piperacillin-tazobactam, and other antipseudomonal beta-lactams purchased/1,000 patient-days were compared during the 2 full calendar years before the antimicrobial restriction program (1995-1996) with the 4 full calendar years after the program was implemented (1998-2001). Pseudomonas aeruginosa resistance trends for the antipseudomonal beta-lactams, ciprofloxacin, and tobramycin also were compared for the 2 years before the program (1995-1996) with the last 2 years of the program (2000-2001). A 44% reduction in ceftazidime use was documented; ostensibly, minimal changes occurred in the overall use of piperacillin and piperacillin-tazobactam. During the same time period, ceftazidime resistance fell from 24% to 11.8% (p<0.001), whereas piperacillin resistance fell from 32.5% to 18.5% (p<0.001). Imipenem resistance declined from 20.5% to 12.3% (p<0.001) with an 18% reduction in use. Aztreonam resistance declined from 29.5% to 16.5% (p<0.001) despite a 57% increase in use. No changes in resistance to either ciprofloxacin or tobramycin were found. CONCLUSION: Through an antimicrobial restriction program, a dramatic reduction in ceftazidime use was achieved with judicious use of other antipseudomonal antimicrobials, which resulted in reduced resistance of P aeruginosa to other beta-lactams. PMID- 12741437 TI - Treating dyslipidemic patients with lipid-modifying and combination therapies. AB - Updated guidelines from the National Cholesterol Education Program give greater emphasis to lipoproteins other than low-density lipoprotein cholesterol (LDL) than previous guidelines. Although statins remain first-line therapy for most patients to lower LDL, combination therapy is the next logical step in achieving goals in patients with mixed dyslipidemia or elevated LDL despite statin therapy. As the prevalence of diabetes, metabolic syndrome, and atherogenic dyslipidemia rises, the importance of treating the total lipid profile becomes even more crucial. Niacin, fibrates, and bile acid sequestrants are effective in combination with statins in lowering LDL, triglycerides, and total cholesterol levels and increasing high-density lipoprotein cholesterol (HDL). Although combination therapies may increase the risk of myopathy, both fibrate-statin and niacin-statin combinations are considered safe. In addition, niacin-statin therapy reduces atherosclerotic progression and coronary events. New pharmacologic formulations exist that will further affect treatment: a single tablet combination of lovastatin and extended-release niacin is available, as is ezetimibe, a cholesterol-absorption inhibitor. In all, both HDL and triglyceride levels correlate with cardiovascular risk and should be considered secondary targets of therapy. Combination therapy can be safe and effective and can be constructed to affect all lipoprotein parameters. PMID- 12741438 TI - Targeting bacterial virulence: the role of protein synthesis inhibitors in severe infections. Insights from the Society of Infectious Diseases Pharmacists. AB - Morbidity and mortality due to certain bacterial pathogens have not declined despite the availability of effective antimicrobial treatments. Staphylococcus aureus and Streptococcus pyogenes cause a number of serious infections, such as necrotizing fasciitis and toxic shock syndrome, which are associated with the release of bacterial toxins. Animal studies have demonstrated clindamycin, a protein synthesis inhibitor, to be more effective in treating these severe infections than other more susceptible antimicrobial treatments. Linezolid, another protein synthesis inhibitor, also has shown efficacy in in vitro studies. Human trials to validate the effects of antibiotic therapies on bacterial virulence have not been performed. Future animal and human studies are needed to help elucidate the immunomodulatory mechanisms of protein synthesis inhibitors in order to optimize antimicrobial treatment and decrease the morbidity and mortality associated with severe bacterial infections. PMID- 12741439 TI - Mississippi mud no more: cost-effectiveness of pharmacokinetic dosage adjustment of vancomycin to prevent nephrotoxicity. AB - OBJECTIVE: To determine the cost-effectiveness of pharmacokinetic dosage adjustment of vancomycin to prevent nephrotoxicity. An analysis was performed for subpopulations of patients receiving nephrotoxic agents (aminoglycosides, amphotericin, and acyclovir), those in the intensive care unit, and those on the oncology service. METHODS: Decision analysis was used to model the cost effectiveness of pharmacokinetic dosage adjustment of vancomycin. The reference case was determined, in part, by a retrospective review of 200 patients randomly selected from our clinical pharmacology consultation service. Patients were aged 18 years or older and had received intravenous vancomycin for at least 48 hours, with at least two--one peak and one trough--vancomycin serum concentrations obtained during therapy. Results of published clinical trials were used to determine the probability of vancomycin-induced nephrotoxicity. RESULTS: The mean cost of treating nephrotoxicity was 11,233 dollars at our institution. The mean cost for all patients was 25,166 dollars (sensitivity analysis 15,000-27,500 dollars)/nephrotoxic episode prevented. The subgroup analysis revealed a cost of 8,363 dollars (sensitivity analysis 4,368-10,500 dollars)/nephrotoxic episode prevented in intensive care patients, 5,000 dollars (sensitivity analysis 1,687 13,250 dollars ) in oncology patients, and a dominant strategy showing a cost savings of 5,564 dollars (sensitivity analysis 2,724-12,428 dollars) in those receiving concomitant nephrotoxins. CONCLUSION: Although pharmacokinetic monitoring and dosage adjustment are effective methods for reducing the toxicity of many drugs, controversy exists regarding the necessity of such monitoring with vancomycin. Evaluation by decision analysis over a range of assumptions, varying probabilities, and costs reveals that pharmacokinetic monitoring and vancomycin dosage adjustment to prevent nephrotoxicity are not cost-effective for all patients. However, such dosage adjustment demonstrates cost-effectiveness for patients receiving concomitant nephrotoxins, intensive care patients, and probably oncology patients. PMID- 12741440 TI - Prescribing patterns and outcomes of enoxaparin for anticoagulation of atrial fibrillation. AB - STUDY OBJECTIVE: To determine prescribing patterns and clinical outcomes of enoxaparin for anticoagulation of atrial fibrillation. DESIGN: Retrospective analysis. SETTING: A 650-bed, tertiary care, community teaching hospital. SUBJECTS: Two hundred thirteen patients who received enoxaparin for thromboprophylaxis during an episode of atrial fibrillation. Intervention. Data collection on demographics, antithrombotic usage, and thrombotic and bleeding episodes from January-June 2001. MEASUREMENTS AND MAIN RESULTS: Patients were characterized as having acute (51.6%) or chronic (48.4%) atrial fibrillation and were categorized according to stroke risk. Three enoxaparin dosing strategies had been prescribed: therapeutic, prophylactic, or adjusted. Prescribed regimens did not reflect stroke risk or type of atrial fibrillation but did reflect the degree of renal impairment. No episodes of stroke occurred with therapeutic enoxaparin dosages, but five strokes occurred among patients receiving prophylactic or adjusted dosages. Bleeding was similar with all dosing strategies in patients with adequate renal function and appeared to be more frequent in those with renal impairment. CONCLUSION: At a single hospital, wide variation in enoxaparin prescribing patterns existed. Further study is necessary to elucidate more fully the appropriate dosing strategy for this agent in the treatment of atrial fibrillation. PMID- 12741442 TI - Smoking-cessation services in Iowa community pharmacies. AB - STUDY OBJECTIVE: To examine community pharmacy practice with regard to providing smoking-cessation counseling. DESIGN: Mailed survey. SETTING: Iowa community pharmacies. PARTICIPANTS: A stratified random sample of pharmacists statewide. MEASUREMENTS AND MAIN RESULTS: Descriptive statistics were computed for all study variables. Fisher exact test or chi2 analysis was performed on selected variables to determine the relationship of each item with pharmacists routinely offering smokers suggestions for quitting. Responses from 129 (38.2%) of 338 pharmacists indicated that although most felt it is important to offer smoking-cessation counseling, about half actually offer this service. Most pharmacists indicated they are prepared to provide counseling, but fewer than 25% had received formal training or were aware of national clinical practice guidelines. Those who had received specific training (p=0.020) or recently attended an educational program (p=0.014) on smoking cessation were more likely to counsel smokers. Primary barriers to providing counseling were lack of time, inability to identify smokers, low patient demand, and lack of reimbursement. CONCLUSION: Our findings suggest that opportunities exist for improving pharmacist education and reducing practice barriers in order to bridge the gap between pharmacists' knowledge and attitudes related to smoking-cessation counseling and their provision of patient counseling in community pharmacy practice. PMID- 12741441 TI - Compliance with clinical practice guidelines for type 2 diabetes in rural patients: treatment gaps and opportunities for improvement. AB - The level of compliance with clinical practice guidelines for patients with type 2 diabetes was evaluated in 368 patients from two health regions in rural northern Alberta, Canada. Data were collected from patient interviews, drug histories, physical and laboratory assessments, and a self-report questionnaire to assess clinical status, indicators of diabetes management, and health care utilization. Treatment of three clinical indicators of diabetes--hemoglobin A1c (A1C), blood pressure, and low-density lipoprotein cholesterol (LDL)--has been shown to reduce the morbidity and mortality associated with type 2 diabetes. Mean +/- SD values for this cohort of patients were as follows: A1C 7.25% +/- 1.54%, blood pressure 131.7 +/- 18.2/76.2 +/- 12.7 mm Hg, and LDL 105.2 +/- 32 mg/dl. Despite these results, only 10.4% of the patients met all three recommended targets for control of glycemia: A1C below 7%, blood pressure below 130/85 mm Hg, and LDL below 100 mg/dl. Of patients not at target levels, 14.4%, 27.5%, and 86.7% reported receiving no therapy for hyperglycemia, hypertension, and dyslipidemia, respectively. Of those taking oral hypoglycemic agents who were not at target levels, only 35% were receiving combination therapy. Of patients at or above LDL target levels, 87% were not receiving any therapy. Only 22% of patients were taking aspirin, although this therapy would be recommended for the entire cohort according to clinical practice guidelines. Despite the availability of proved effective therapies, treatment gaps were present for this cohort of patients. PMID- 12741443 TI - Purple toes syndrome associated with warfarin therapy in a patient with antiphospholipid syndrome. AB - Purple toes syndrome is an extremely uncommon, nonhemorrhagic, cutaneous complication associated with warfarin therapy. It is characterized by the sudden appearance of bilateral, painful, purple lesions on the toes and sides of the feet that blanch with pressure. The syndrome usually develops 3-8 weeks after the start of warfarin therapy. A 47-year-old man with a history of purple toes syndrome that resolved after discontinuing warfarin--prescribed for a deep vein thrombosis (DVT) in his right lower leg--experienced an acute, proximal DVT in his other leg. Warfarin again was prescribed; 1 week later, purple toes syndrome developed in that extremity. Warfarin therapy again was discontinued, and intravenous unfractionated heparin was started; the patient's clinical picture indicated a possible pulmonary embolism, and laboratory analysis suggested antiphospholipid syndrome. The patient's toe pain resolved, but the purple discoloration persisted. Follow-up laboratory analysis confirmed antiphospholipid syndrome, and warfarin was restarted with close monitoring. No further complications occurred with long-term therapy. Although a rare complication of therapy, clinicians should monitor for the development of purple toes syndrome in patients taking warfarin. PMID- 12741444 TI - Shock-like sensations during venlafaxine withdrawal. AB - Electric shock-like sensations may occur after cessation of treatment with serotonin selective reuptake inhibitors but are reported in the literature only rarely with discontinuation of venlafaxine. Two patients experienced severe shock like sensations during venlafaxine withdrawal. For both patients symptoms occurred with lowering of the dosage and persisted for 5 days after complete discontinuation of the drug. These sensations may represent significant alteration of neuronal activity in the central nervous system. PMID- 12741445 TI - Tooth discoloration after treatment with linezolid. AB - With the increasing frequency of methicillin-resistant Staphylococcus aureus in immunocompromised hosts, clinicians are increasingly prescribing the oral treatment option of linezolid. Linezolid is the first of a new class of antibiotics, the oxazolidinones. The drug is generally well tolerated. However, mild-to-moderate adverse effects have been reported, such as gastrointestinal effects (most frequent), myelosuppression, skin eruptions, elevated liver enzymes, and tongue discoloration. As with any new drug on the commercial market, not all adverse effects are elucidated during preclinical trials. An immunocompromised 11-year-old girl with cellulitis of the toe experienced tooth discoloration after receiving a 28-day course of linezolid. The discoloration was present on the enamel of her lower anterior teeth and was superficial and reversible with dental cleaning. PMID- 12741446 TI - Validity of the general practice research database. AB - The United Kingdom General Practice Research Database (GPRD) is an office-based, computer-generated, medical resource designed from its inception to be used for epidemiologic research. A distinct version of the GPRD is maintained by the Boston Collaborative Drug Surveillance Program and has been the source of more than 130 scientific articles primarily addressing drug safety issues. We reviewed evidence related to the validity of the GPRD. Specifically, with our extensive experience with this automated database, we evaluated the quality and completeness of the data that it contains. PMID- 12741447 TI - Dose conversion from epoetin alfa to darbepoetin alfa for patients with chronic kidney disease receiving hemodialysis. PMID- 12741448 TI - Helicobacter pylori is not and never was "protective" against anything, including GERD. PMID- 12741450 TI - Significance of an exaggerated meal-stimulated gastrin response in pathogenesis of Helicobacter pylori-negative duodenal ulcer. AB - The aim of this study was to clarify the pathogenesis of Helicobacter pylori negative duodenal ulcer (DU) by investigating the meal-stimulated serum gastrin (SG) response. The subjects were 9 patients with H. pylori-negative DU, 28 H. pylori-positive DU, 11 H. pylori-positive volunteers, and 30 H. pylori-negative volunteers. Blood samples were taken before and after consumption of a test meal. The integrated 1-hr gastrin response (IGR) was taken to be the area under the SG time curve, calculated by the trapezoid method. H. pylori infection status was determined by histology, serology, and the [13C] urea breath test. The mean basal SG concentration was lower in the H. pylori-negative DU patients than in the H. pylori-positive DU patients, but an exaggerated IGR was observed in three patients (33.3%) with H. pylori-negative DU. In conclusion, our findings indicate that an exaggerated meal-stimulated gastrin response may contribute to the pathogenesis of H. pylori-negative DU. PMID- 12741449 TI - Helicobacter pylori stimulates inducible nitric oxide synthase in diverse topographical patterns in various gastroduodenal disorders. AB - Overproduction of nitric oxide by inducible nitric oxide synthase (iNOS) acts cytotoxically and contributes to inflammation. We explored the roles of iNOS in the pathogenesis of Helicobacter pylori-associated diseases. Using reverse transcribed PCR, we examined topographical patterns of iNOS mRNA expression in the gastroduodenal mucosa in H. pylori-negative controls and H. pylori-positive patients with duodenal ulcer (DU), gastric ulcer (GU), and ulcer-free gastritis. iNOS expression showed topographical variations among the tested disorders. As compared to controls, DU had a significantly higher expression of iNOS mRNA in the duodenum, GU in the antrum and duodenum, and gastritis in the antrum and corpus. H. pylori eradication yielded a significant reduction of iNOS mRNA in the duodenum of DU and in the antrum of GU. Diverse topographical patterns of H. pylori-induced iNOS expression may contribute to mechanisms by which H. pylori elicits different clinical disorders. PMID- 12741451 TI - A randomized, double-blind comparison of two different coffee-roasting processes on development of heartburn and dyspepsia in coffee-sensitive individuals. AB - The mechanism underlying coffee-induced heartburn and dyspepsia remains poorly understood. This has led to speculation that variations in coffee processing may be important. The primary objective of this study was to determine whether a coffee brewed with coffee beans processed using conduction roasting will result in fewer symptoms of gastroesophageal reflux and dyspepsia in coffee-sensitive individuals compared to a differently processed yet otherwise similar coffee. Thirty coffee-sensitive individuals completed this single-center, randomized, double-blind, crossover study in which the symptoms of heartburn, regurgitation and dyspepsia were assessed following coffee consumption both in the fasting state and after ingestion of a standard test meal. Consumption of both coffees resulted in heartburn, regurgitation, and dyspepsia in most individuals. No significant differences in the frequency or severity of heartburn, regurgitation, or dyspepsia were demonstrated between the two coffees either in the fasting state or after the test meal. We conclude that differences in the coffee bean roasting process do not result in marked differences in coffee-induced upper gastrointestinal symptoms. PMID- 12741452 TI - Reductions in symptom distress reported by patients with moderately severe, nonerosive gastroesophageal reflux disease treated with rabeprazole. AB - Gastroesophageal reflux disease (GERD) is characterized by heartburn and related symptoms that are distressing to patients and interfere with everyday functioning and well-being. A measure of symptom distress, the GERD Symptom Assessment Scale (GSAS), was included in two randomized, placebo-controlled trials of rabeprazole among patients with nonerosive GERD. The age (mean +/- SD) of the 223 patients was 43.5 +/- 11.9 years, and most were female (67%) and Caucasian (78%). Significantly greater reductions in symptom distress were observed among patients receiving rabeprazole 20 mg daily for 4 weeks relative to those receiving placebo (-0.62 vs -0.36, P < 0.0001). The magnitude of this treatment difference was comparable to the differences observed between levels of overall symptom improvement on the patient global rating (0.2 and 0.3 points; P < 0.0001). In conclusion, reducing symptom distress is an important goal of therapeutic interventions for GERD. Rabeprazole significantly reduced the distress associated with a broad range of GERD symptoms, and the magnitude of this effect was meaningful to patients. PMID- 12741453 TI - Role of endogenous endothelin-1 in ethanol-induced gastric mucosal damage in humans. AB - Gastric microcirculatory disturbances are involved in the ethanol-induced gastric mucosal damage. In this study in humans we evaluated the time course of plasma and gastric mucosal endothelin-1 (ET-1) concentrations after intragastric ethanol administration; furthermore we determined the correlation among changes in gastric tissue endothelin-1 and microscopic and gross gastric hemorrhagic damage. ET-1 concentrations in plasma and gastric mucosa were measured by radioimmunoassay. The endoscopic appearance of the gastric mucosa was evaluated and scored on a scale of 0-5, and gastric biopsies for histological evaluation were obtained from the antral and the corpus mucosa just before and 30 min after 40% ethanol administration in seven healthy volunteers. Plasma ET-1 concentration increased as soon as 20 min after ethanol administration, reached a significant peak at 30 min (P < 0.01), and returned to near basal level within 120 min. Gastric mucosal ET-1 concentration significantly increased 30 min after ethanol administration in both the body (P < 0.05) and the antrum (P < 0.05) of the stomach; however the ET-1 increase was significantly higher in the body. Moreover, data obtained 30 min after ethanol administration showed a significant correlation between gastric mucosal ET-1 levels and gross hemorrhagic damage (r = 0.84). A significant correlation was also observed between antral gastric mucosal ET-1 and microscopic lesions (r = 0.70). We conclude that 40% ethanol, given orally, stimulates the release of gastric mucosal endothelin-1 and causes a rapid and time-dependent increase of ET-1 plasma level in humans. The increased plasma and gastric tissue endothelin-1 concentration may play a role in ethanol-induced gastric hemorrhagic injury in humans. PMID- 12741454 TI - Expression of Bcl-2 family modulated through p53-dependent pathway in human hepatocellular carcinoma. AB - The biological behavior of hepatocellular carcinoma (HCC) is, in part, relevant to apoptosis. A systematic investigation of the apoptosis-related Bcl-2 family modulated by p53 in HCC is lacking. A total of 22 HCC patients were studied. The expression of p53 protein in HCC was assessed using the immunohistochemical method, which categorized the HCC patients into two groups: group 1, with immunonegative p53 (N = 7); and group 2, with immunopositive p53 (N = 15). The expression of p53, p21, Bcl-2, Bax, Bcl-xL, and Bcl-xS in the 22 HCC cases detected by western bioting was quantified with a densitometer. The apoptosis of the 22 HCC cases was determined by terminal deoxynucleotidyltransferase-mediated UTP end-labeling (TUNEL). We found that Bcl-2 was remarkably up-regulated in group 2 (14 of 15), but was down-regulated in group 1 (5 of 7). Bax was up regulated in both group 1 (6 of 7) and group 2 (13 of 15). Bcl-xL was up regulated in both group 1 (5 of 7) and group 2 (9 of 15). Bcl-xS was remarkably down-regulated in group 2 (14 of 15) compared to group 1 (4 of 7). The apoptosis indexes of groups 1 and 2 were 0.82 +/- 0.26% and 0.33 +/- 0.17%, respectively (P = 0.023). The long-term survival of group 1 was superior to that of group 2 (log rank test, P = 0.001). In conclusion, Bcl-2 and Bcl-xS represented the most significant anti- and proapoptotic proteins, respectively, modulated through a p53-dependent pathway in HCC. PMID- 12741456 TI - A rare case of simultaneous gastric adenocarcinoma and squamous cell carcinoma of the esophagus in a Peruvian, geriatric, female. PMID- 12741455 TI - Clinical manifestations of primary hepatic angiosarcoma. AB - Malignant tumors of the liver stemming from mesenchymal origins are rare neoplasms, <1% of primary malignant hepatic lesions. Primary hepatic angiosarcoma (PHA) is the most common (36%). This study describes the incidence and clinical characteristics of this rare tumor in two medical centers, over the past 18 years. We reviewed tumor registry files at Jackson Memorial Hospital and oncology data records at Cedar's Medical Center, 1979-1997. A total of 865 primary hepatic tumors were identified, of which five cases (0.58%) were PHA; four were men, and the median age was 53 years. Symptoms and signs included: pain, anemia, fever of unknown origin, weight loss, abdominal mass, and hemoperitoneum. Median survival was only 6 months. In conclusion, primary hepatic angiosarcomas frequently are symptomatic. The presentation and preexisting factors are valuable in establishing a clinical suspicion to diagnose this rare tumor. Although imaging studies are helpful, they are not conclusive, and liver biopsy is usually required. PMID- 12741457 TI - Involvement of primary afferent nerves after abdominal irradiation: consequences on ileal contractile activity and inflammatory mediator release in the rat. AB - In this study we analyzed the role of substance P (SP) from afferent nerves in ileum contractibility and in the release of inflammatory mediators (neurotensin, Il-1beta, and TNF-alpha) in ileal mucosa and muscularis layers after a 10-Gy gamma-irradiation of the abdomen. Six hours after irradiation, SP concentrations were lower than in control rats, and 3 days after irradiation SP-induced contractile activity was higher. Irradiation significantly increased the levels of neurotensin, Il-1beta, and TNF-alpha in both layers. Pretreatment with capsaicin depleted afferent nerve endings of SP and reduced SP levels by about 50%. Capsaicin treatment reduced SP concentrations further, beyond the levels due to irradiation, thereby suggesting that all sources of SP are affected by irradiation. Capsaicin treatment prevented the irradiation from affecting SP induced contractile response or increasing neurotensin levels. This finding suggests that SP released by afferent nerve endings controls these functions. Proinflammatory cytokine release was not reduced by capsaicin treatment. PMID- 12741458 TI - Therapeutic potential of gastric electrical stimulation for obesity and its possible mechanisms: a preliminary canine study. AB - Our aim was to investigate the effects of gastric electrical stimulation on food intake, weight, gastric myoelectrical, and parasympathetic activity. Dogs were implanted with serosal electrodes and a subcutaneous stimulator. The stimulator was turned on and off alternately every month for 4 months. Food intake, weight, gastric myoelectrical activity, and electrocardiograms were recorded. Daily food intake and weight were significantly decreased during the months with stimulation. Stimulation did not show any acute effect on gastric myoelectrical activity; however, it chronically and significantly impaired gastric myoelectrical activity in the fed state, but not in the fasting state. The parasympathetic activity in the fasting state assessed from the spectral analysis of heart rate variability was markedly decreased with stimulation both acutely and chronically. In conclusion, chronic gastric electrical stimulation results in a reduction in food intake, weight loss, a reduction in parasympathetic activity, and chronic inhibition of gastric myoelectrical activity. These data suggest that gastric electrical stimulation is a potential therapy for the treatment of obesity and its inhibitory effect on food intake and weight may involve both muscles and the vagal afferent pathway. PMID- 12741459 TI - Effect of hypo- and hyperthyroidism on gastric myoelectrical activity. AB - Although hypo- and hyperthyroid patients have different symptoms in the gastrointestinal tract, the mechanism of thyroid action on the gut remains poorly understood. Thus the aim of this study was to investigate the effect of hypo- and hyperthyroidism on gastric myoelectrical activity, gastric emptying, dyspeptic symptoms. Twenty-two hyperthyroid (median age 45, 15 females) and 11 hypothyroid (median age 42, 10 females) patients were included into the study. Dyspepsia score, hypo- and hyperthyroid symptom scale, abdominal ultrasonography and upper gastrointestinal endoscopy were performed. Gastric myoelectrical activity was measured by electrogastrograpy (EGG) before and after therapy both preprandially and postprandially and compared with age, gender, and body-matched controls (12 for hypothyroid, 15 for hyperthyroid patients). Radionuclide gastric emptying studies were performed with a solid meal. Hypothyroid patients revealed a significant increase in preprandial tachygastria as compared with controls (12.3% vs 4.8%). The percentage of preprandial normal slow waves (2.4-3.7 cpm) was below 70% (dysmotility) in 7 of 11 hypothyroid patients versus 2 of 12 controls (P < 0.05). Hyperthyroid patients revealed a significantly higher preprandial (3.1 vs 2.8) and postprandial (3.4 vs 3) DF when compared with the controls (P < 0.05). A higher percentage of postprandial taschygastria (7.9 vs 0) was present in hyperthyroid patients than in the controls (P < 0.05). The decrease on postprandial EGG power (power ratio < 1) was observed in 7 patients the in hyperthyroid group and 1 in controls (P < 0.05). The percentage of postprandial normal slow waves was below 70% in 10 of 20 hyperthyroid patients vs 1 of 15 controls (P < 0.05). After therapy these differences disappeared in the euthyroid state. The hypo- and hyperthyroid symptom scale correlated to dyspepsia score. Dyspepsia score in hyperthyroidism correlated to power ratios in hyperthyroid patients. We detected some correlations between serum levels of fT3 or fT4 and some EGG parameters in hypo- and hyperthyroidism. Dyspepsia score and hypo- and hyperthyroid symptom scale were improved significantly after therapy in the euthyroid state. In conclusions, we showed gastric dysrhythmia by EGG in both hypo- and hyperthyroid patients. Dyspeptic symptoms correlated to the activity of thyroid disease. After therapy, these findings and dyspeptic symptoms improved in the euthyroid state. Abnormalities of power ratios may be responsible of dyspeptic symptoms in hyperthyroid patients. EGG may be a useful and noninvasive tool for detecting gastric disturbances during hypo- and hyperthyroidism. PMID- 12741460 TI - Microendoscopic nasointestinal feeding tube placement in mechanically ventilated patients with gastroparesis. AB - Gastroparesis often precludes gastric enteral nutrition (EN) in critically ill patients. Our aim was to determine the feasibility of bedside microendoscopic placement of nasointestinal feeding tubes to facilitate enteral nutrition in critically ill patients with poor gastric emptying. Nine mechanically ventilated patients with proven gastroparesis underwent 10 nasointestinal intubations using a microendoscope. These were compared with 35 patients who underwent pH sensor guided intubation. Blind pH-guided intubation was faster than microendoscopic placement (21.4 +/- 10.7 v 32 +/- 11.6 min, P = 0.016) and cheaper in terms of disposables [87 pounds sterling (132 dollars) vs 222 pounds sterling (337 dollars) per intubation, P < 0.0001]. Depth of placement (postpyloric: 64% vs 50% including 32% vs 50% reaching duodenum part 3, 4, or jejunum, both NS) was similar. We conclude that microendoscopy failed to improve transpyloric intubation due to poor visualization of gastrointestinal anatomy and difficulty maneuvering the tube-endoscope ensemble. However, when successful, transpyloric placement was always deep, permitting immediate and full EN. To date, the technique and equipment is not superior to pH-guided placement and is not suitable for use by personnel with minimal training. PMID- 12741461 TI - Up-regulation of p75 neurotrophin receptor (p75NTR) is associated with apoptosis in chronic pancreatitis. AB - Activation of apoptosis in chronic pancreatitis has been demonstrated. The low affinity neurotrophin receptor p75 (p75NTR) mediates apoptosis in many cell types in vivo and in vitro. The aim of this study was to examine whether p75NTR is involved in the apoptotic process in chronic pancreatitis. The quantity and localization of the receptor was evaluated using northern blot analysis, in situ hybridization, immunohistochemistry, and western blot analysis. Apoptosis was determined by TUNEL assay. By northern blot analysis, p75NTR mRNA levels were increased 40-fold in chronic pancreatitis compared with normal pancreas (P < 0.01). In situ hybridization revealed weak p75NTR mRNA expression in some ductal cells in the normal pancreas. In contrast in chronic pancreatitis moderate p75NTR expression was present in acinar cells next to fibrosis, ductal cells, and cells of ductular structures as well as in some islet cells. Immunostaining of p75NTR in normal pancreas and chronic pancreatitis tissue samples showed a similar intensity and distribution pattern as found by in situ hybridization. Higher p75NTR protein levels could be confirmed by western blot analysis, which revealed an 8.6-fold increase of p75NTR in chronic pancreatitis. TUNEL staining showed, in chronic pancreatitis samples, positivity in some acinar cells next to fibrosis, some ductal cells, and cells of ductular structures. Also some islet cells were positive by TUNEL staining. The presence of p75NTR immunoreactivity was positively correlated (P < 0.05) with the apoptotic index in the exocrine and endocrine pancreas. In conclusion, p75NTR, the low-affinity receptor of neurotrophins which mediates apoptosis, is up-regulated in CP and is involved in the apoptotic process of the exocrine and endocrine pancreas. PMID- 12741462 TI - Adenomyomatosis of the gallbladder in a child. PMID- 12741463 TI - Intestinal obstruction following biliopancreatic diversion. AB - In our opinion biliopancreatic diversion patients require particular care, especially in the emergency room setting. Due to the changes in anatomy following surgery, there are many changes in physiology. These changes result in different clinical manifestations than in the general population. Therefore, the combined use of clinical and laboratory findings, abdominal ultrasound and CT scan are invaluable assets in obesity surgery patients presenting with acute abdominal pain. PMID- 12741464 TI - Ischemic-appearing papillitis in a patient with gallstone pancreatitis and cholangitis. PMID- 12741465 TI - Extraintestinal symptoms in irritable bowel syndrome and inflammatory bowel diseases: nature, severity, and relationship to gastrointestinal symptoms. AB - Patients suffering from the irritable bowel syndrome (IBS) tend to have extraintestinal symptoms. The purposes of this study were to compare the nature and severity of these symptoms in IBS patients in relation to patients with inflammatory bowel disease (IBD) and to nonpatients and to clarify the relationship between intestinal and extraintestinal symptoms. A consecutive group of male patients and a control group of age-matched male subjects were studied. Symptoms were graded for severity using a validated, self-administered inventory. There were 53 IBS patients, 55 IBD patients (32 Crohn's disease), and 56 controls. IBS patients scored significantly higher than IBD patients on constipation, dyspepsia, and reflux scales. Musculoskeletal symptoms, neurasthenia, and sleep scores were similar in IBS and IBD patients, and both groups scored significantly higher than the controls. The scores of urinary, thoracic, and oral symptoms were similar in IBD patients and in controls. However, IBS patients scored significantly higher than both groups on all these scales. Reflux symptoms were the most powerful predictors of extraintestinal symptoms, both in IBS and in IBD. Diarrhea was predictive of extraintestinal symptoms only in IBD. In conclusion, IBS patients experienced extraintestinal symptoms to the same extent, or even more than patients with IBD. However, the relationship between intestinal and extraintestinal symptoms differed in the two conditions. PMID- 12741466 TI - Granulocyte adsorptive apheresis for pediatric patients with ulcerative colitis. AB - Granulocytapheresis (GCAP) has produced efficacy in adult patients with ulcerative colitis (UC) by adsorbing activated granulocytes and monocytes/macrophages. We retrospectively investigated efficacy and safety of GCAP in pediatric patients with active UC. Twelve steroid-refractory children (12.2 +/- 3.1 years old) were treated with GCAP, one session/week for 5-10 consecutive weeks. In 8 patients, clinical symptoms improved after two GCAP sessions. Normal body temperature, stool frequency, and disappearance of blood in stool were seen after 24.3 +/- 11.5 days. The endoscopic grade improved from 2.6 +/- 0.3 to 0.4 +/- 0.2. One patient who initially responded, developed bloody diarrhea later and 2 cases remained unchanged. The dose of steroid was tapered during GCAP therapy by 50%. No serious adverse effects were noted. Four of 8 cases relapsed 3.5 +/- 2.2 months after the last GCAP while on maintenance therapy, the other 4 were in remission up to 22.8 +/- 18.1 months. In conclusion, GCAP appears to be effective and well tolerated in children with steroid refractory UC. PMID- 12741467 TI - Gum arabic (GA) modifies paracellular water and electrolyte transport in the small intestine. AB - Previous experiments have shown that a soluble fiber, gum arabic (GA), enhances water, electrolyte, and glucose absorption in animal models of diarrhea. The mechanisms implicated in this effect have not been fully elucidated. This study examined the possibility that paracellular transport is modulated by luminal GA, resulting in an enhanced rate of absorption in the small intestine. This hypothesis was tested by 3-hr jejunal perfusions on anesthetized rats with solutions containing 140 mM NaCl, 5 mM KCl, and 2 microCi/liter (74 kBq) 3H2O, with either 2.5 g/liter GA [+GA] or in its absence [CTL], and one of the following agents, capable of altering paracellular transport: chenodeoxycholic acid at 0.5 mM (CDC), 2,4,6-triaminopyrimidine (TAP) at 20 mM, and protamine at 100 mg/liter (PTM). Sodium, potassium, net water, and unidirectional water movement were measured. The addition of GA increased sodium absorption in perfusions with CDC, TAP, or PTM only. Similar effects by GA on net water absorption rates were obtained in tissues permeabilized with CDC and PTM; however, GA added to TAP did not normalize the reduction caused by TAP. Although PTM did not alter net water absorption, addition of GA to perfusates with PTM enhanced absorption values above those of CTL. GA reversed the strong negative effects of CDC on potassium absorption but was ineffective in this regard with TAP and PTM. The data obtained with those reagents that affect paracellular transport and the histological evidence support the view that GA promotes net absorption by this route in the small intestine of normal rats. PMID- 12741468 TI - Presentations of adult celiac disease in a nationwide patient support group. AB - Recent epidemiological studies primarily from Europe document that adult celiac disease often lacks the classic presentation of steatorrhea and weight loss. There are few surveys of adult celiac disease in the United States. We surveyed the large population of a nationwide patient support group to determine their disease presentations. In the initial survey (N = 1032 respondents), the median age at onset was 46 years, and the diagnosis of adult celiac disease was often delayed (median 12 months, with 21% delayed over 10 years). Only 32% of adults were underweight, and only about 50% reported frequent diarrhea and weight loss. A second survey documented that common presenting symptoms were fatigue (82%), abdominal pain (77%), bloating or gas (73%), and anemia (63%). Initial physician diagnoses were often irritable bowel syndrome (37%), psychological disorders (29%), and fibromyalgia (9%). These initial presentations are similar to those in Europe and often resemble irritable bowel syndrome. PMID- 12741469 TI - Effects of incremental starvation on gut mucosa. AB - Starvation induces gut mucosal atrophy, but the effects of progressive dietary restriction are not defined. The study's purpose was to determine the effects of incremental starvation on gut epithelial cell turnover. After food intake of mice was determined, they were divided into five groups: control (ad libitum fed), 75% normal intake, 50% intake, 25% intake, and fasted. Mice were killed after 48 hours, and the proximal small bowel were assessed for weight and protein content. Histologic specimens were examined for villus morphology, apoptosis, and proliferation. After 48 hr of diet restriction, bowel weight decreased in the 50% intake, 25% intake, and fasted groups. Villus density also decreased in the fasted group. Proliferation progressively decreased in the diet-restricted groups. Apoptosis increased in the fasted group, primarily in the villus tip. In conclusion, incremental starvation produces progressive small bowel atrophy. The mechanism involves both decreased gut epithelial cell proliferation and increased apoptosis. PMID- 12741470 TI - Dose-response relationship and mechanism of action of Saccharomyces boulardii in castor oil-induced diarrhea in rats. AB - For biotherapeutic agents, there is a lack of information on dose-response relationships and mechanism of action. The present study was designed to address these issues for Saccharomyces boulardii using the rat model of castor oil induced diarrhea. A single dose of Saccharomyces boulardii at 12 x 10(10) CFU/kg of viable cells given 1 hr before castor oil administration significantly reduced the onset of diarrhea. Repeated ingestion of the yeast, twice daily between 1.2 and 12 x 10(10) CFU/kg for 5 days before castor oil, showed a dose-response relationship. The percentage of rats with diarrhea decreased and a stronger protection was afforded by the repeated treatment. The mechanism of action of Saccharomyces boulardii in this model was investigated with two classes of antagonists, naloxone and L-arginine. The effect of Saccharomyces boulardii was not inhibited by naloxone but was significantly reduced by L-arginine. This last result suggests a novel mechanism of action for Saccharomyces boulardii involving a possible inhibition of nitric oxide production by the yeast. PMID- 12741471 TI - Time course of colonic nuclear factor-kappa B expression during bacterial peptidoglycan-polysaccharide-induced colitis in rats. AB - Nuclear factor-kappa B has been proposed to play a role in the pathogenesis of bacterial peptidoglycan-polysaccharide-induced colitis. However, its colonic expression has not been defined in detail. The primary aim of this study was to profile this expression in the rat colon. Peptigoglycan-polysaccharide was administered to the rat colon by direct intramural injections. Gross colonic injury was determined at various time points. Concomitantly, colonic nuclear factor-kappa B was measured by an electrophoretic mobility shift assay and by immunohistochemistry. Gross colonic injury and colonic nuclear factor-kappa B expression showed similar time courses following peptigoglycan-polysaccharide administration. Peak colonic injury and nuclear factor-kappa B expression were found on day 21. Nuclear factor-kappa B was mainly expressed in submucosal inflammatory cells. In conclusion, the administration of bacterial peptidoglycan polysaccharide to the rat colon caused a chronic colitis, which was characterized by up-regulated colonic nuclear factor-kappa B. PMID- 12741472 TI - Linear IgA bullous dermatosis: an association with ulcerative colitis versus renal cell carcinoma. PMID- 12741473 TI - Effect of protein kinase C activation and inhibition on rat hepatic stellate cell activation. AB - Protein kinase C (PKC) may play a role in the intracellular signaling pathways responsible for transforming hepatic stellate cells into myofibroblasts. This study examined the effects of inhibitors and activators of PKC on hepatic stellate cell activation. Stellate cells isolated from normal rats were incubated with either 10(-5) M chelerythrine, 10(-7) M bisindolylmaleimide I hydrochloride (BIM), or 10(-6) M staurosporine (PKC inhibitors), or 10(-7) M phorbol myristate acetate (PMA) or 10(-6) M thymeleatoxin (PKC activators). Chelerythrine suppressed alpha-smooth muscle actin expression and proliferation by 49% and 33%, respectively. BIM inhibited alpha-smooth muscle actin expression by 60%, but had no significant effect on proliferation. Staurosporine decreased proliferation by 86% and completely prevented alpha-smooth muscle actin expression. PKC activators had divergent effects on proliferation and alpha-smooth muscle actin expression. PMA and thymeleatoxin caused a 2.8- to 3.2-fold increase in proliferation, while suppressing alpha-smooth muscle actin expression by 50-70%. The demonstration that hepatic stellate cell activation can be suppressed by PKC inhibitors suggests a role for PKC in the regulation of hepatic stellate cell activation. PMID- 12741474 TI - Prevalence of transaminase abnormalities in asymptomatic, healthy subjects participating in an executive health-screening program. AB - The objective of this study was to characterize the prevalence of asymptomatic liver transminase (LT) abnormalities in a healthy, low-risk adult population and identify associated risk factors. We reviewed 2340 completed medical records of participants in our Executive Health Program, which provided screening medical evaluations for executives. LT (alanine aminotransferase and aspartate aminotransferase) were considered abnormal if they above normal range for our laboratory. Subjects were excluded if they had a history of viral hepatitis, nonviral liver disease, or an identifiable cause of LT elevation. Of the 2340 subjects 2294 met inclusion criteria and all had AST recorded, but only 1309 had ALT recorded. In all, 341 subjects (14.9%) were found to have abnormal LT and in those who had less than 3 drinks per day, 13.9% had elevated LT and 3.6% had LT twice the upper limit of normal. Of the 1309 subjects in whom both AST and ALT were measured, 20.8% had abnormal LT and 6.3% had LT twice the upper limit of normal. On univariate analysis age < 60 (P = 0.005), male sex (P < 0.0001), body mass index > or = 30 (P < 0.0001), cholesterol > or = 200 mg/dl (P = 0.018), and triglycerides > or = 200 mg/dl (P < 0.0001) were associated with abnormal LT; all these variables except cholesterol were significant by logistic regression analysis. The odds ratio of abnormal LT and LT 2 times normal was 1.79 (CI 1.20 2.68) and 2.50 (CI 1.04-6), respectively, in subjects with one risk factor, and 2.80 (CI 1.07-7.34) and 4.73 (CI 0.91-24.5), respectively, in subjects with four risk factors. In conclusion, there is a high prevalence of LT abnormalities in this healthy population. Subjects with multiple risk factors should be considered for screening. PMID- 12741475 TI - Helicobacter pylori infection is associated with greater impairment of cytochrome P-450 liver metabolic activity in anti-HCV positive cirrhotic patients. AB - Helicobacter pylori gastric infection has been associated with various digestive and extradigestive diseases. In liver disease bacterial infections have been associated with impairment of cytochrome P-450 liver metabolic activity. Moreover, infection by Helicobacter spp. seems to be linked with the development of hepatocellular carcinoma (HCC) in mice. Our aims were to evaluate the influence of H. pylori infection on cytochrome P-450 liver metabolic activity as assessed by means of monoethylglycinexylidide (MEGX) test and to assess the prevalence of H. pylori infection in patients with HCC. Ninety-six hepatitis C virus (HCV) -positive cirrhotic patients, 36 of whom had HCC, were tested for H. pylori infection by means of anti-H. pylori IgG. Patients underwent the MEGX test. Characteristics of the patients were then analyzed on the basis of the presence of H. pylori infection. Seroprevalence of H. pylori infection was similar between cirrhotic patients without (68%) or with (63.8%) HCC. Mean MEGX values were significantly (P < 0.0001) lower in H. pylori infected patients (18.2 +/- 13.9 ng/ml) as compared to the noninfected ones (46.9 +/- 17.1 ng/ml), independently of Child-Pugh's classification. These differences persisted even after subdividing patients according to the presence of HCC. In conclusion, in anti-HCV positive cirrhotic patients H. pylori infection is associated to an impairment of cytochrome P-450 liver metabolic activity. Seroprevalence of H. pylori infection in HCC patients is similar to that observed in tumor-free cirrhotics. PMID- 12741476 TI - Nonresponse to interferon monotherapy in HCV-related chronic hepatitis: results of retreatment and prognostic factors. AB - At present two of the most relevant problems of the therapy of HCV-related chronic active hepatitis are the retreatment of nonresponders to interferon-alpha (IFN-alpha) and the definition of a prognostic index of response. We treated 44 patients who previously were nonresponders to IFN-alpha alone with IFN-alpha plus ribavirin for 12 months. Among the tests performed, we included the serum level of soluble intercellular adhesion molecule-1 (sICAM-1) at the beginning of the trial and at 3 months thereafter. We obtained 56.81% end of treatment responses and 47.72% sustained responses. A decrease of at least 10% of the sICAM-1 serum level during the first 3 months of treatment strongly correlated to the results of therapy while the usual important prognostic factors (HCV genotype and viral load) did not show this relation. PMID- 12741477 TI - Risk factors for hepatitis C virus infection among patients receiving health care in a Department of Veterans Affairs hospital. AB - A cross-sectional, seroepidemiological study was conducted to determine the prevalence and risk factors for hepatitis C virus (HCV) infection among veterans receiving health care from the VA. Among 274 evaluated outpatients, anti-HCV was found in 27 (9.9%). The prevalence of anti-HCV was 3.7% among 190 individuals who reported no illicit drug use compared to 24.7% among 81 subjects who had used drugs (P < 0.001). The prevalence of anti-HCV was 4.8% among 208 veterans who had never been incarcerated compared to 27.9% among 61 veterans who had been incarcerated (P < 0.001). A multivariate model found the following factors to be independently associated with anti-HCV; having used illicit drugs [odds ratio (OR) = 3.7, 95% CI 1.3-11.8; P = 0.001), having been incarcerated (OR = 4.4, 95% CI 1.7-10.9; P = 0.001), and a yearly income less than 10,000 US dollars (OR = 3.5, 95% CI 1.3-9.4; P = 0.002). Because HCV infection was most strongly associated with illicit drug use, incarceration, and low income, these risk factors should be utilized to develop screening strategies among VA patients. PMID- 12741478 TI - Hyperammonemic encephalopathy in a patient with ureterosigmoidostomy and acute hepatitis: a specific case of fulminant hepatic failure. PMID- 12741480 TI - Transverse myelitis occurring in association with primary biliary cirrhosis and Sjogren's syndrome. AB - Transverse myelitis (TM) as a manifestation of an autoimmune disorder is relatively rare. In Sjogren's syndrome (SS), the occurrence of TM is remarkably uncommon. Only three cases have been reported associated with primary biliary cirrhosis (PBC). Here we report the fourth case of TM occurring in association with SS and PBC. Patients with unexplained transverse myelitis require a careful search for an underlying etiology to include the findings of SS and PBC. The precise pathogenesis of TM in patients with SS is unknown. Most show good response to steroids. Cyclophosphamide and chlorambucil may be useful in those who respond poorly to steroids. PMID- 12741479 TI - Effects of quercetin on liver damage in rats with carbon tetrachloride-induced cirrhosis. AB - Flavonoids are reported to exhibit a wide variety of biological effects, including antioxidant and free radical-scavenging activities. Evidence of oxidative reactions is often associated with various chronic disease processes characterized by accumulation of connective tissue. This study was aimed to investigate the protective effects of chronic administration of the flavonoid quercetin (150 micromol/kg body wt/day intraperitoneally) in rats with carbon tetrachloride-induced fibrosis. In animals rendered cirrhotic by administration of carbon tetrachloride for 16 weeks, cell necrosis, fibrosis, and inflammatory infiltration were found. Histological abnormalities were accompanied by a higher hepatic content of collagen and thiobarbituric acid-reactive substances. Expression of inducible nitric oxide synthase (iNOS) was significantly increased in the liver. Treatment with quercetin during 3 weeks improved liver histology and reduced collagen content, iNOS expression, and lipid peroxidation. Those effects were associated with an increased total peroxyl radical-trapping antioxidant capacity of liver. We conclude that quercetin is effective in this model of liver damage. PMID- 12741483 TI - Applications of 1 MV field-emission transmission electron microscope. AB - A newly developed 1 MV field-emission transmission electron microscope has recently been applied to the field of superconductivity by utilizing its bright and monochromatic field-emission electron beam. This microscope allows individual magnetic vortices inside high-Tc superconductors to be observed, thus, opening the way to investigate the unusual behaviour of vortices, which reflects the anisotropic layered structure of these superconducting materials. One example is the observation of the arrangements of chain vortex lines that are formed when a magnetic field is applied obliquely to the layer plane of the materials. PMID- 12741481 TI - Pylephlebitis of portal venous system associated with urinary infection. PMID- 12741482 TI - A spherical aberration-corrected 200 kV TEM. AB - A spherical aberration (Cs)-corrected 200 kV TEM was newly developed. The column of the microscope was extended by 25 cm and the inner yoke of the objective lens was modified to insert some parts of the corrector elements. The corrector has two hexapole elements that play a main role in Cs correction and they are placed at a position equivalent to the coma-free point of the objective lens by using two transfer doublet lenses. The Cs correction was successfully carried out by means of the third-order aberration that was generated in the two extended hexapoles. The Cs can be corrected to the desired value and also can be overcompensated in order to produce a negative Cs, as with the corrected Cs of 23 microm shown in this work. The optical system of the corrector does not produce second- and fourth-order aberrations, and can correct residual aberrations up to the third order. All of the corrector elements are computer controlled and the third-order aberrations are quite stable after they are properly corrected. The resolution of 0.135 nm was experimentally confirmed by the Young's fringe method. Image simulations of a silicon [110] single crystal were made with various Cs and defocus values to demonstrate the effectiveness of arbitral control of Cs. PMID- 12741484 TI - Electron holographic 3-D nano-analysis of Au/TiO2 catalyst at interface. AB - Three-dimensional (3-D) nanostructures of gold catalysts supported on TiO2 were analysed by electron holography and high-resolution electron microscopy. The contact angle of the gold particle on TiO2 tended to be >90 degrees in the case of gold particles with a size (height) of >4 nm and it tended to be <90 degrees for gold particles with a height of <2 nm. The change in morphology increases the perimeter at the Au/TiO2 interface as the particle size decreases. This change in 3-D structure should be attributed to a change in electronic structure at the interface. It was found that electron holography enabled 3-D analysis at the atomic level and was effective for analysing nanostructured particles. PMID- 12741485 TI - ARHVTEM of the Pd/ZnO heterointerface chemical structure. AB - Atomic-resolution high-voltage transmission electron microscopy was applied to the atomic structure analysis of a well-defined Pd/ZnO polar interface, which was produced by internal oxidation. Viewing the ZnO along <1120> axis, each atomic column consists of either oxygen or zinc, so that an individual column was directly identified from the image contrast of the picture taken at the Scherzer defocus condition. The terminating chemical element at the Pd/ZnO interface, which is parallel to [111]Pd and to [0001]ZnO, was shown to be zinc but not oxygen on both sides of the precipitate. PMID- 12741487 TI - Influence of Sr-doping in Ba7Rh6O18, a new one-dimensional oxide of the homologous series (A3Rh2O6)alpha(A3Rh3O9)beta. AB - The synthesis and microstructural characterization, by means of selected-area electron diffraction and high-resolution electron microscopy, of Ba7Rh6O18 is reported. This material, isostructural to Ba7Co6O18, is formed by rows of one trigonal prism and five face-sharing octahedra running parallel to the c-axis of a trigonal unit cell with parameters a = 1.004(5) and c = 3.165(7) nm. The substitution of Sr by Ba is accommodated by means of twin formation due to a rearrangement of the rows of polyhedra. PMID- 12741486 TI - Atomistic observation and simulation analysis of spatio-temporal fluctuations during radiation-induced amorphization. AB - We performed a dynamical-atomistic study of radiation-induced amorphization in the NiTi intermetallic compound using in situ high-resolution high-voltage electron microscopy and molecular dynamics simulations in connection with image simulation. Spatio-temporal fluctuations as non-equilibrium fluctuations in an energy-dissipative system, due to transient atom-cluster formation during amorphization, were revealed by the present spatial autocorrelation analysis. PMID- 12741488 TI - High-resolution ultrahigh-vacuum electron microscopy of helical gold nanowires: junction and thinning process. AB - Helical multishell (HMS) gold nanowires were observed in situ by ultra-high vacuum electron microscopy. During thinning of the helical nanowire, a junction was formed between two nanowires of different diameter. The structure of the gold junction is proposed in comparison with the multiwall carbon nanotube. The gold junction has a dislocation-like core to accommodate the number difference of the atomic rows that compose the HMS nanowires, while the carbon junction has a pair of five- and seven-membered rings. Thinning of the helical gold nanowire by electron-beam radiation can be caused by the climbing motion of the dislocation like core along the nanowire. PMID- 12741489 TI - In situ HREM study on the structural instability of isolated nanometre-sized alloy particles in the Sn-Bi system. AB - The structural instability of isolated nm-sized alloy particles has been investigated by in situ transmission electron microscopy, using particles in the Sn-Bi system. In a pure tin (Sn) particle, no structural fluctuation was induced under electron-beam irradiation. In a tin-rich solid solution particle, an orientational fluctuation took place at a rate of approximately once per 1-3 s. In a high concentration alloy particle with a two-phase microstructure, a structural fluctuation occurred at a rate of a few hertz. Namely, the fluctuation became more frequent with increasing bismuth (Bi) concentration, no matter whether it consists of a single phase or multiple phases. A good parallelism can be found between this fluctuation enhancement with bismuth concentration and the fact that the free-energy difference between a solid particle and the corresponding liquid one decreases continuously with bismuth concentration and approaches a value close to zero at the eutectic composition. These results lead to a view that a nm-sized solid particle exhibits a structural instability under electron-beam irradiation when the free-energy difference between a solid particle and the corresponding liquid one is reduced to a value close to zero. PMID- 12741490 TI - Radiation-induced segregation and precipitation behaviours around cascade clusters under electron irradiation. AB - We have investigated the formation of cascade clusters and structural changes in them by means of electron irradiation following ion irradiation in an austenitic stainless steel. Almost all of the cascade clusters, which were introduced by the ion irradiation, grew to form interstitial-type dislocation loops or vacancy-type stacking fault tetrahedra after electron irradiation at 623 K, whereas a few of the dot-type clusters remained in the matrix. It was possible to recognize the concentration of Ni and Si by radiation-induced segregation around the dot-type clusters. After electron irradiation at 773 K, we found that some cascade clusters became precipitates (delta-Ni2Si) due to radiation-induced precipitation. This suggests that the cascade clusters could directly become precipitation sites during irradiation. PMID- 12741491 TI - First observation of SiO2/Si(100) interfaces by spherical aberration-corrected high-resolution transmission electron microscopy. AB - SiO2/Si(100) interfaces were for the first time observed by a spherical aberration-corrected high-resolution transmission electron microscope in a cross sectional mode. As the Fresnel fringes were not contrasted at the interfaces, the interfacial structures were clearly observed without the need for artificial image contrast. Atomic steps and defects on the Si(100) surfaces were accurately identified. Also, image simulations with the target imaging performance revealed oxygen atomic columns between silicon-silicon bonds. The present instrument is of potential use for semiconductor science and technology, even for the analysis of oxygen atoms at interfaces. PMID- 12741492 TI - Creep deformation of grain boundary in a highly crystalline SiC fibre. AB - Silicon carbide (SiC) matrix composites reinforced by SiC fibres (SiC/SiC composites) are currently being considered as alternative materials in high Ni alloys for high-temperature applications, such as aerospace components, gas turbine energy-conversion systems and nuclear fusion reactors, because of their high specific strength and fracture toughness at elevated temperatures compared with monolithic SiC ceramics. It is important to evaluate the creep properties of SiC fibres under tensile loading in order to determine their usefulness as structural components. However, it would be hard to evaluate creep properties by monoaxial tensile properties when we have little knowledge on the microstructure of crept specimens, especially at the grain boundary. Recently, a simple fibre bend stress relaxation (BSR) test was introduced by Morscher and DiCarlo to address this problem. Interpretation of the fracture mechanism at the grain boundary is also essential to allow improvement of the mechanical properties. In this paper, effects of stress applied by BSR test on microstructural evolution in advanced SiC fibres, such as Tyranno-SA including small amounts of Al, are described and discussed along with the results of microstructure analysis on an atomic scale by using advanced microscopy. PMID- 12741493 TI - In situ HREM observation of nucleation and growth of nanotwins beneath the solid liquid interface in Si. AB - Nucleation and growth of nanotwins in Si grown from Al-Si liquid have been observed directly using an in situ heating experiment in a high-resolution transmission electron microscope. Nanotwins are nucleated at the triple point between a vacuum and the solid-liquid interface. When two parallel twins, the mirror planes of which are separated slightly, encounter each other, very complicated atomic arrangements are formed. The structure of the perturbed region is discussed tentatively in terms of the high-pressure phases in Si. PMID- 12741494 TI - Inhomogeneous substitution of polyhalogenated copper-phthalocyanine studied by high-resolution imaging and electron crystallography. AB - The present study was aimed at distinguishing halogen atoms substituted in a molecule of copper-phthalocyanine (CuPc) by high-resolution transmission electron microscopy (HRTEM). The subject was carried out on octabromooctachloro-CuPc and obtained HRTEM images suggested inhomogeneous distribution of bromine and chlorine atoms at 1-, 2-, 3- and 4-substitution positions of CuPc in image contrast. As a result of electron crystallography, the occupancy of bromine atoms at the 1-position (and equally at the 4-position) was found to be 52%, which is slightly higher than for random substitution. This small deviation causes apparent inhomogeneous image contrast of halogen substitution observed in the high-resolution images, as concluded from computer simulation of images. Such potential to detect inhomogeneity in substitution by TEM should be important for the chemical understanding of substitution reactions at the atomic level. PMID- 12741495 TI - The design and implementation of a motion correction scheme for neurological PET. AB - A method is described to monitor the motion of the head during neurological positron emission tomography (PET) acquisitions and to correct the data post acquisition for the recorded motion prior to image reconstruction. The technique uses an optical tracking system, Polaris, to accurately monitor the position of the head during the PET acquisition. The PET data are acquired in list mode where the events are written directly to disk during acquisition. The motion tracking information is aligned to the PET data using a sequence of pseudo-random numbers, which are inserted into the time tags in the list mode event stream through the gating input interface on the tomograph. The position of the head is monitored during the transmission acquisition, and it is assumed that there is minimal head motion during this measurement. Each event, prompt and delayed, in the list mode event stream is corrected for motion and transformed into the transmission space. For a given line of response, normalization, including corrections for detector efficiency, geometry and crystal interference and dead time are applied prior to motion correction and rebinning in the sinogram. A series of phantom experiments were performed to confirm the accuracy of the method: (a) a point source located in three discrete axial positions in the tomograph field of view, 0 mm, 10 mm and 20 mm from a reference point, (b) a multi-line source phantom rotated in both discrete and gradual rotations through +/- 5 degrees and +/- 15 degrees, including a vertical and horizontal movement in the plane. For both phantom experiments images were reconstructed for both the fixed and motion corrected data. Measurements for resolution, full width at half maximum (FWHM) and full width at tenth maximum (FWTM), were calculated from these images and a comparison made between the fixedand motion corrected datasets. From the point source measurements, the FWHM at each axial position was 7.1 mm in the horizontal direction, and increasing from 4.7 mm at the 0 mm position, to 4.8 mm, 20 mm offset, in the vertical direction. The results from the multi-line source phantom with +/- 5 degrees rotations showed a maximum degradation in FWHM, when compared with the stationary phantom, of 0.6 mm, in the horizontal direction, and 0.3 mm in the vertical direction. The corresponding values for the larger rotation, +/- 15 degrees, were 0.7 mm and 1.1 mm, respectively. The performance of the method was confirmed with a Hoffman brain phantom moved continuously, and a clinical acquisition using [11C]raclopride (normal volunteer). A visual comparison of both the motion and non-motion corrected images of the Hoffman brain phantom clearly demonstrated the efficacy of the method. A sample time-activity curve extracted from the clinical study showed irregularities prior to motion correction, which were removed after correction. A method has been developed to accurately monitor the motion of the head during a neurological PET acquisition, and correct for this motion prior to image reconstruction. The method has been demonstrated to be accurate and does not add significantly to either the acquisition or the subsequent data processing. PMID- 12741496 TI - An improved analytical detector response function model for multilayer small diameter PET scanners. AB - The optimization of spatial resolution is a critical consideration in the design of small-diameter positron emission tomography (PET) scanners for animal imaging, and is often addressed with Monte Carlo simulations. As a faster and simpler solution, we have developed a new analytical model of the PET detector response function, and implemented the model for a small single-slice, multilayer PET scanner. The accuracy of the model has been assessed by comparison with both Monte Carlo simulations and experimental measurements published in the literature. Results from the analytical model agreed well with the Monte Carlo method, being noise free and two to three orders of magnitude faster. The only major discrepancy was a slight underestimation of the width of the point spread function by the analytical method as inter-crystal scatter is neglected. We observed good agreement between the predictions of the model and experimental measurements. For two large-diameter scanners additional discrepancies were seen due to photon acollinearity, which is not considered in the model. We have shown that the simple and fast analytical detector response function model can provide accurate estimates of spatial resolution for small-diameter PET scanners, and could be a useful tool for several applications, complementing or cross validating other simulation methods. PMID- 12741497 TI - Evaluation of penetration and scattering components in conventional pinhole SPECT: phantom studies using Monte Carlo simulation. AB - In quantitative pinhole SPECT, photon penetration through the collimator edges (penetration), and photon scattering by the object (object scatter) and collimator (collimator scatter) have not been investigated rigorously. Monte Carlo simulation was used to evaluate these three physical processes for different tungsten knife-edge pinhole collimators using uniform, hotspot and donut phantoms filled with 201Tl, 99mTc, 123I and 131I solutions. For the hotspot phantom, the penetration levels with respect to total counts for a 1 mm pinhole aperture were 78%, 28% and 23% for 131I, 123I and 99mTc, respectively. For a 2 mm aperture, these values were 65% for 131I, 16% for 123I and 12% for 99mTc. For all pinholes, 201Tl penetration was less than 4%. The evaluated scatter (from object and collimator) with a hotspot phantom for the 1 mm pinhole was 24%, 16%, 18% and 13% for 201Tl, 99mTc, 123I and 131I, respectively. Summation of the object and collimator scatter for the uniform phantom was approximately 20% higher than that for the hotspot phantom. Significant counts due to penetration and object and collimator scatter in the reconstructed image were observed inside the core of the donut phantom. The collimator scatter can be neglected for all isotopes used in this study except for 131I. Object scatter correction for all radionuclides used in this study is necessary and correction for the penetration contribution is necessary for all radionuclides but 201Tl. PMID- 12741498 TI - An analytical image reconstruction algorithm to compensate for scattering angle broadening in Compton cameras. AB - Compton cameras have been developed for use in gamma-ray astronomy and nuclear medicine. Their defining merit is that they do not need collimators; however, on the demerit side, they need inversion procedures for image reconstruction, since a measured datum is proportional to the integration of incident gamma rays along a cone surface with the same Compton scattering angle. First, an iteration method was adopted for this task. Later, analytical methods were found under restricted conditions. Parra (2000 IEEE Trans. Nucl. Sci. 47 1543-50) deduced a purely analytical reconstruction algorithm for a complete set of scattering-projection data that include data at all the scattering angles. Tomitani and Hirasawa (2002 Phys. Med. Biol. 47 2129-45) found that by making a slight modification, Parra's algorithm could be extended to the scattering-projection data in limited scattering angles. However, their algorithm neglected the effects of practical problems that cause the degradation of spatial resolution. Sources of degradation were identified as noise in the energy signal of their front detector and the Doppler effect in the scattering process. In this paper, we first analyse the effects of these sources on the angular resolution of the scattering-projection data and then present a revised reconstruction algorithm in which these two factors are incorporated. Simulation studies on digital phantoms reveal that the algorithm can reconstruct images even when these two factors are included. PMID- 12741499 TI - 1H MRS of a boron neutron capture therapy 10B-carrier, L-p-boronophenylalanine fructose complex, BPA-F: phantom studies at 1.5 and 3.0 T. AB - The quantification of a BNCT 10B-carrier, L-p-boronophenylalanine-fructose complex (BPA-F), was evaluated using 1H magnetic resonance spectroscopy (1H MRS) with phantoms at 1.5 and 3.0 T. For proper quantification, relaxation times T1 and T2 are needed. While T1 is relatively easy to determine, the determination of T2 of a coupled spin system of aromatic protons of BPA is not straightforward with standard MRS sequences. In addition, an uncoupled concentration reference for aromatic protons of BPA must be used with caution. In order to determine T2, the response of an aromatic proton spin system to the MRS sequence PRESS with various echo times was calculated and the product of the response curve with exponential decay was fitted to the measured intensities. Furthermore, the response curve can be used to correct the intensities, when an uncoupled resonance is used as a concentration reference. BPA was quantified using both phantom replacement and internal water referencing methods with accuracies of +/- 5% and +/- 15%. Our phantom results suggest that in vivo studies on BPA concentration determination will be feasible. PMID- 12741500 TI - Feasibility of salvage interstitial microwave thermal therapy for prostate carcinoma following failed brachytherapy: studies in a tissue equivalent phantom. AB - Thermal therapy is an experimental treatment to destroy solid tumours by heating them to temperatures ranging from 55 degrees C to 90 degrees C, inducing thermal coagulation and necrosis of the tumour. We are investigating the feasibility of interstitial microwave thermal therapy as a salvage treatment for prostate cancer patients with local recurrence following failed brachytherapy. Due to the electrical and thermal conductivity of the brachytherapy seeds, we hypothesized that the seeds could scatter the microwave energy and cause unpredictable heating. To investigate this, a 915 MHz helical antenna was inserted into a muscle-equivalent phantom with and without brachytherapy seeds. Following a 10 W, 5 s input to the antenna, the temperature rise was used to calculate absorbed power, also referred to as specific absorption rate (SAR). Plane wave models based on Maxwell's equations were also used to characterize the electromagnetic scattering effect of the seeds. In addition, the phantom was heated with 8 W for 5 min to quantify the effect of the seeds on the temperature distribution during extended heating. SAR measurements indicated that the seeds had no significant effect on the shape and size of the SAR pattern of the antenna. However, the plane wave simulations indicated that the seeds could scatter the microwave energy resulting in hot spots at the seed edges. Lack of experimental evidence of these hot spots was probably due to the complex polarization of the microwaves emitted by the helical antenna. Extended heating experiments also demonstrated that the seeds had no significant effect on the temperature distributions and rates of temperature rise measured in the phantom. The results indicate that brachytherapy seeds are not a technical impediment to interstitial microwave thermal therapy as a salvage treatment following failed brachytherapy. PMID- 12741501 TI - A CT calibration method based on the polybinary tissue model for radiotherapy treatment planning. AB - A method to establish the relationship between CT number and effective density for therapeutic radiations is proposed. We approximated body tissues to mixtures of muscle, air, fat and bone. Consequently, the relationship can be calibrated only with a CT scan of their substitutes, for which we chose water, air, ethanol and potassium phosphate solution, respectively. With simple and specific corrections for non-equivalencies of the substitutes, a calibration accuracy of 1% will be achieved. We tested the calibration method with some biological materials to verify that the proposed method would offer the accuracy, simplicity and specificity required for a standard in radiotherapy treatment planning, in particular with heavy charged particles. PMID- 12741502 TI - Polymer gel measurement of dose homogeneity in the breast: comparing MLC intensity modulation with standard wedged delivery. AB - Polymer gel dosimetry has been used to measure the radiotherapy dose homogeneity in a breast phantom for two different treatment methods. The first 'standard' method uses two tangential wedged fields while the second method has three static fields shaped by multileaf collimators (MLCs) in addition to the standard wedged fields to create intensity modulated fields. Gel dose distributions from the multileaf modulation treatment show an improved dose uniformity in comparison to the standard treatment with a decreased volume receiving doses over 105%. PMID- 12741503 TI - Inverse planning for intensity-modulated arc therapy using direct aperture optimization. AB - Intensity-modulated arc therapy (IMAT) is a radiation therapy delivery technique that combines gantry rotation with dynamic multi-leaf collimation (MLC). With IMAT, the benefits of rotational IMRT can be realized using a conventional linear accelerator and a conventional MLC. Thus far, the advantages of IMAT have gone largely unrealized due to the lack of robust automated planning tools capable of producing efficient IMAT treatment plans. This work describes an inverse treatment planning algorithm, called 'direct aperture optimization' (DAO) that can be used to generate inverse treatment plans for IMAT. In contrast to traditional inverse planning techniques where the relative weights of a series of pencil beams are optimized, DAO optimizes the leaf positions and weights of the apertures in the plan. This technique allows any delivery constraints to be enforced during the optimization, eliminating the need for a leaf-sequencing step. It is this feature that enables DAO to easily create inverse plans for IMAT. To illustrate the feasibility of DAO applied to IMAT, several cases are presented, including a cylindrical phantom, a head and neck patient and a prostate patient. PMID- 12741504 TI - Comparison of biologically damaging spectral solar ultraviolet radiation at a southern hemisphere sub-tropical site. AB - The first dataset of a complete year of biologically damaging spectral UV at a sub-tropical latitude in the southern hemisphere has been presented. The new data provides a baseline dataset against which comparisons can be made in the future to establish if there have been any long term trends in the biologically damaging UV. The general shape of the variation of the daily biologically damaging exposures through the year depends on the relative response of the various action spectra at the different wavelengths. The ratio of the daily erythemal to actinic exposures drops by approximately 20 to 25% from winter to summer. The ratio of the erythemal to DNA exposures drops by approximately 50% over the same period. In contrast, the ratio of the erythemal to plant damage exposures is higher in summer compared to winter. This is due to the changes in the relative proportion of UVA to UVB wavebands and relative responses of the different action spectra. The relative changes for the different action spectra show that the erythemal action spectrum cannot be used as a proxy for other biologically damaging responses. PMID- 12741505 TI - A novel image quality index using Moran I statistics. AB - Measurement of image quality is very important for various applications such as image compression, restoration and enhancement. Conventional methods (e.g., mean squared error; MSE) use error summation to measure quality change pixel by pixel and do not correlate well with subjective quality measurement. This is due to the fact that human eyes extract structural information from the viewing field. In this study a new quality index using a Moran I statistics is proposed. The Moran statistic that measures the sharpness from a local area is a good index of quality as most image processing techniques alter the smoothness of the image. Preliminary results show that the new quality index outperforms the MSE significantly under various types of image distortions. PMID- 12741506 TI - X-ray energy choice for lung tumour irradiation depends on the density distribution of clonogenic cells. PMID- 12741508 TI - Host oyster tissue extracts modulate in vitro protease expression and cellular differentiation in the protozoan parasite, Perkinsus marinus. AB - Perkinsus marinus is responsible for a chronic disease (Dermo) of the Eastern oyster, Crassostrea virginica. In order to simulate the in vivo environment more closely, a chemically defined medium (JL-ODRP-3) was supplemented with tissue homogenate extracts or plasma from oysters possessing varying degrees of susceptibility to P. marinus infection. In media supplemented with extracts from highly susceptible oysters (C. virginica), P. marinus cells secreted elevated amounts of a set of low molecular weight serine proteases (LMP: 30-45 kDa) as assessed by enhanced digestion within gelatin-substrate SDS-PAGE gels. Oyster species of low susceptibility (C. gigas and C. ariakensis) did not exhibit this ability to upregulate P. marinus LMP expression. Oyster extract supplementation also led to pronounced changes in P. marinus cellular morphology, such that the cells were comparable to those observed within naturally infected oysters. PMID- 12741507 TI - Efficacy comparison of intravenous artelinate and artesunate in Plasmodium berghei-infected Sprague-Dawley rats. AB - This paper reports the comparative antimalarial efficacy of intravenous artelinate and artesunate in rats. Prior to efficacy experiments, a Plasmodium berghei-Sprague-Dawley rat model of malaria was developed, in which the clearance effects of intravenous drugs could be readily compared. In efficacy experiments, groups of P. berghei-infected rats were given 3 daily intravenous treatments of artelinate or artesunate at molar equivalent dose rates (total of 0-191.2 micromoles/kg). Artelinate was superior to artesunate in terms of clearance (100% clearance dose of 95.6 micromoles/kg (40 mg/kg) versus 191.2 micromoles/ kg for AS (73.4 mg/kg)) and parasite clearance time (1.7 +/- 0.5 days for AL versus 2.7 +/- 0.5 days for AS at a dose rate of 191.2 micromoles/kg, P < 0.01). No frank clinical toxicity was observed, though both artesunate and artelinate induced dose-related vascular necrosis at the site of injection. The necrosis was less severe and reversible when the drugs were administered via femoral, rather than tail/foot veins. The data suggest that the P. berghei-7-week-old Sprague-Dawley rat model of malaria is reproducible and useful for assessing the efficacy of antimalarials and that artelinate is at least as potent, and safe, as artesunate, the leading clinical treatment for severe malaria. PMID- 12741509 TI - Vaccination of mice against experimental Neospora caninum infection using NcSAG1- and NcSRS2-based recombinant antigens and DNA vaccines. AB - NcSAG1 and NcSRS2, the two major immunodominant tachyzoite surface antigens of the apicomplexan parasite Neospora caninum, were investigated for their potential as vaccine candidates in mice. Recombinant recNcSRS2 and recNcSAG1 were expressed in Escherichia coli as poly-histidine-tagged fusion proteins. Separate groups of mice were immunized with purified recNcSAG1, recNcSRS2, or a combination of both, and were then challenged with N. caninum tachyzoites. Subsequent experiments included intramuscular vaccination of mice with the eukaryotic expression plasmid pcDNA3 containing either NcSRS2 or NcSAG1 cDNA inserts, followed by a single booster with the corresponding recombinant antigens. Immunization with a crude somatic antigen (NC1-extract) was included in the experiments. Following challenge, the presence of the parasite in the different organs was assessed by a N. caninum-specific PCR, while the parasite burden in infected brain tissue was assessed by quantitative real-time PCR. Immunization of mice employing individual recombinant antigens, or combined recNcSAG1/recNcSRS2, resulted in a lower degree of protection against cerebral infection, when compared to combined DNA/recombinant antigen vaccination. Serological analysis showed that this protective effect was associated with the occurrence of antibodies directed against native parasite antigens in those animals receiving combined DNA/recombinant antigen vaccination. Conversely, mice immunized with recombinant antigens alone generated antibodies recognizing only the recombinant antigens. Mice experiencing clinical signs such as walking disorders, rounded back, apathy and paralysis were observed only in the untreated positive control groups, but never in the vaccinated groups. Our results suggest that a combined DNA/recombinant antigen-vaccine, based on NcSAG1 and NcSRS2, respectively, exhibited a highly significant protective effect against experimentally induced cerebral neosporosis in mice. PMID- 12741510 TI - Light and electron microscopic study of the myxosporean, Henneguya friderici n. sp. from the Amazonian teleostean fish, Leporinus friderici. AB - A new histozoic species of myxosporean was found to infect the gill filaments, gut, kidney and liver of the freshwater teleost Leporinus friderici, collected from the estuarine region of the Amazon, near the city of Belem, Brazil. The plasmodia show asynchronous development, at any one time composed of mature spores and all sporogonic stages. The ellipsoidal spore body, measuring 10.4 microm long and 5.7 microm wide, consists of 2 equal shell valves adhering together along the straight suture line. Each valve has a caudal process measuring 23.3 microm in length. There are 2 symmetric polar capsules, without intercapsular appendix, measuring 5.0 microm x 2.1 microm, and each has a polar filament with 7-8 coils. In general, ultrastructural details of sporoblast and spore development are in agreement with previously described myxosporeans. Some ultrastructural aspects such as cellular alterations of the pericyte in the different organs infected and characterization of the sporoplasmosomes during the sporoplasm maturation are described. This parasite was studied under light and electron microscope and compared with others species of the genus Henneguya, considering also host specificity. From our observations we propose the creation of a new species, Henneguya friderici n. sp. PMID- 12741511 TI - Immunoparasitological parameters of the intestinal phase of trichinellosis in rats. AB - Enzyme-linked immunosorbent assays (ELISAs) were developed in order to detect coproantigens (CAgs), coproantibodies (CAbs) and faecal immune complexes (FIC) in rats experimentally infected with Trichinella spiralis. The usefulness of these assays was compared to that of a conventional ELISA for detection of serum antibodies (Abs) to muscle larvae excretory-secretory products (ML-ESP). The ELISA for CAgs was the first parameter to give a positive result but the detection was limited only to day 2 p.i. CAbs against ML-ESP and adult worm excretory-secretory products (AW-ESP) was first positive on day 4 p.i. Anti-ML ESP remained positive until day 12 p.i. while CAbs against AW-ESP remained positive throughout the study period. Specific IgE and IgA were found. FIC were detected between days 2 and 8 p.i. Serum Abs began to appear on day 10 p.i. Therefore, the ELISA for CAbs was a suitable assay for the detection of the enteral and early phases of the infection. PMID- 12741512 TI - Mating interactions of Schistosoma haematobium and S. intercalatum with their hybrid offspring. AB - Experiments were designed to study the mating behaviour between the Schistosoma haematobium male x S. intercalatumfemale hybrid and the 2 parental species S. haematobium and S. intercalatum. Individual worms were identified by electrophoretic analysis of glucose-6-phosphate dehydrogenase, which was characteristic for each isolate. Analysis of the data obtained showed that both heterospecific and homospecific pairs formed between the hybrids and S. haematobium and S. intercalatum. S. haematobium and the hybrid are better than S. intercalatum in forming pairs, and S. haematobium showed a greater homospecific mate preference compared with the hybrid. Analysis of the data using the Mantel Haenszel test suggests that mating competition does exist between the schistosomes, with the hybrid being dominant over both the parental species and S. haematobium being dominant over S. intercalatum. The hybrid males showed a greater ability than S. intercalatum and S. haematobium males in taking away S. haematobium and S. intercalatum females from their homospecific males when introduced into a pre-established S. haematobium or S. intercalatum infection. They were able to take females from S. intercalatum homospecific pairs more easily compared with females from S. haematobium homospecific pairs. The significance of the results is discussed in relation to the epidemiological changes of schistosomiasis in Cameroon, where hybridization between S. haematobium and S. intercalatum has taken place, with S. haematobium and the hybrid managing to replace the endemic S. intercalatum over the last 30 years. PMID- 12741513 TI - A predator's dilemma: prey choice and parasite susceptibility in three-spined sticklebacks. AB - The acanthocephalan parasite Pomphorhynchus laevis is transmitted by a suitable intermediate host such as the amphipod Gammarus pulex to its definitive fish host. Parasite-induced alterations in both appearance and behaviour concur to render infected gammarids more vulnerable to predation, thus promoting parasite transmission. Experimental infection of laboratory bred full- and half-sib groups of three-spined sticklebacks (Gasterosteus aculeatus) provided evidence that the parasite imposes a survival cost proportional to the severity of infection on its final host. Variation among sibships in the susceptibility to infection was consistent. When given a choice, fish consumed significantly more infected than uninfected prey. Overall, more resistant fish did not prey upon infected gammarids more often than did relatively susceptible fish. Only fish with a relatively high physical condition properly adjusted prey selection to the extent of their parasite susceptibility, thus exploiting the enhanced profitability of infected prey. PMID- 12741514 TI - Cytochemical studies of the neuromuscular systems of the diporpa and juvenile stages of Eudiplozoon nipponicum (Monogenea: diplozoidae). AB - Using indirect immuno- and enzyme-cytochemical techniques, interfaced with confocal scanning laser microscopy and standard optical microscopy, neuronal pathways have been demonstrated in whole-mount preparations of the unpaired diporpae and freshly paired juvenile stages of Eudiplozoon nipponicum (Monogenea: Diplozoidae). All 3 main classes of neuronal mediators, cholinergic, aminergic and peptidergic, were identified throughout both central and peripheral elements of a well-differentiated orthogonal nervous system. Neural mapping revealed considerable overlap and similarity in staining of the nervous systems of the diporpa and adult worm. The main differences in the diporpa relate to the innervation of the temporary ventral sucker and dorsal papilla, structures which are unique to the larva and which enable fusion between worms but then disappear. Branches from the longitudinal nerve cords innervate these structures and appear to be involved in the process of somatic fusion, probably giving rise to the inter-specimen connections that later link the 2 central nervous systems in paired adult parasites. In the hindbody, there is extensive haptoral innervation associated with the developing clamps and small central hooks. Reactive neuronal components were found associated with the early stages of clamp development prior to connections being made with the extrinsic adductor muscle bundles. The muscle systems of the diporpa and juvenile stages comprise a lattice-like arrangement of circular, longitudinal and diagonal fibres that make up the body wall, together with buccal suckers, haptoral clamps and associated adductor muscles, and the transient ventral sucker. All have obvious importance to diporpae when they migrate over the gill and undertake body contact, torsion and fusion during the process of pairing. Behaviour during the pairing of diporpae is described. PMID- 12741515 TI - Effects of experimental Schistocephalus solidus infections on growth, morphology and sexual development of female three-spined sticklebacks, Gasterosteus aculeatus. AB - The use of naturally infected hosts in studies attempting to identify parasite induced changes in host biology is problematical because it does not eliminate the possibility that infection may be a consequence, rather than a cause, of host trait variation. In addition, uncontrolled concomitant infections may confound results. In this study we experimentally infected individual laboratory-bred female three-spined sticklebacks Gasterosteus aculeatus L. with the pseudophyllidean cestode Schistocephalus solidus [Muller], and compared the morphology and growth patterns of infected females with sham-exposed controls over a 16-week period. Fish were fed a ration of 8% body weight per day. Non invasive image analysis techniques allowed the growth of individual plerocercoids to be tracked in vivo throughout the course of infection, and patterns of host and parasite growth were determined. Females that developed infections diverged morphometrically from unexposed control females and exposed-uninfected females at 6 weeks post-infection, with the width of the body at the pectoral fins giving the earliest indication of infection success. When including the plerocercoid, infected females gained weight more quickly than controls, but when plerocercoid weight was removed this trend was reversed. There was no effect of infection on the increase in fish length. Plerocercoids grew at different rates in individual hosts, and exhibited measurable sustained weight increases of up to 10% per day. Final estimates of plerocercoid weight from morphometric analysis prior to autopsy were accurate to within +/- 17% of actual plerocercoid weight. At autopsy, infected female sticklebacks had significantly lower perivisceral fat reserves but had developed significantly larger ovaries than controls. The results are discussed in relation to previous studies examining natural infections, and the value of utilizing experimental infections to examine ecological aspects of host-parasite interactions is discussed. PMID- 12741516 TI - The consequences of self-fertilization and outcrossing of the cestode Schistocephalus solidus in its second intermediate host. AB - Many hermaphroditic parasites reproduce by both cross-fertilization and self fertilization. To understand the maintenance of such mixed mating systems it is necessary to compare the fitness consequences of the two reproductive modes. This has, however, almost never been done in the context of host-parasite coevolution. Here we show the consequences of outcrossing and selfing in an advanced life stage of the cestode Schistocephalus solidus, i.e. in its second intermediate host, the three-spined stickleback (Gasterosteus aculeatus). Each juvenile stickleback was simultaneously exposed to 2 experimentally infected copepods, one harbouring outcrossed the other selfed parasites. At 60 days p.i. parasites were removed from the fish's body cavity and, with microsatellite markers, assigned to either outcrossed or selfed origin. Prevalence was not significantly higher in outcrossed parasites. However, those fish that were infected contained significantly more out-crossed than selfed parasites. Thus the probability of a selfed parasite to progress in the life-cycle is reduced in the second intermediate host. Furthermore, we found that even the multiply infected fish increased in weight during the experiment. Nevertheless, total worm weight in multiply infected fish was significantly lower than in singly infected ones, which thus might be a parasite life-history strategy. PMID- 12741517 TI - Habitat overlap and gastrointestinal parasitism in sympatric African bovids. AB - Gastrointestinal parasite infections are widespread among wild ungulates. Because many of these parasites infect multiple host species, inter-specific interactions among hosts potentially play an important role in parasite transmission dynamics in ungulate communities. In this study, the effects of inter-specific contact on parasitism rates in 11 sympatric African bovids was examined using habitat overlap among species as a measure of cross-species contact rates. Across individual hosts, strongyle nematode abundance increased with increasing numbers of bovid species occupying a habitat. Furthermore, comparative analyses show a positive association between strongyle prevalence and level of habitat overlap across taxa. These findings suggest that among sympatric bovids, contact between species contributes significantly to the transmission of generalist nematode parasites. For a more host-specific parasite group, coccidia, parasite abundance and individual probability of infection declined in hosts living in bovid rich habitats. This pattern may reflect enhanced interspecific competition among parasites in these areas. Finally, similar to strongyle abundance, individual parasite richness also increased among hosts occupying habitats with higher numbers of bovid species. No association between habitat overlap and parasite richness was detected at higher taxonomic scales, however, which suggests that contact between host species may not contribute to parasite colonization of new host taxa. PMID- 12741518 TI - A critical evaluation of the role of alternative oxidase in the performance of strobilurin and related fungicides acting at the Qo site of complex III. AB - Mitochondrial respiration conserves energy by linking NADH oxidation and electron coupled proton translocation with ATP synthesis, through a core pathway involving three large protein complexes. Strobilurin fungicides block electron flow through one of these complexes (III), and disrupt energy supply. Despite an essential need for ATP throughout fungal disease development, strobilurins are largely preventative; indeed some diseases are not controlled at all, and several pathogens have quickly developed resistance. Target-site variation is not the only cause of these performance difficulties. Alternative oxidase (AOX) is a strobilurin-insensitive terminal oxidase that allows electrons from ubiquinol to bypass Complex III. Its synthesis is constitutive in some fungi but in many others is induced by inhibition of the main pathway. AOX provides a strobilurin insensitive pathway for oxidation of NADH. Protons are pumped as electrons flow through Complex I, but energy conservation is less efficient than for the full respiratory chain. Salicylhydroxamic acid (SHAM) is a characteristic inhibitor of AOX, and several studies have explored the potentiation of strobilurin activity by SHAM. We present a kinetic-based model which relates changes in the extent of potentiation during different phases of disease development to a changing importance of energy efficiency. The model provides a framework for understanding the varying efficacy of strobilurin fungicides. In many cases, AOX can limit strobilurin effectiveness once an infection is established, but is unable to interfere significantly with strobilurin action during germination. A less stringent demand for energy efficiency during early disease development could lead to insensitivity towards this class of fungicides. This is discussed in relation to Botrytis cinerea, which is often poorly controlled by strobilurins. Mutations with a similar effect may explain evidence implicating AOX in resistance development in normally well-controlled plant pathogens, such as Venturia inaequalis. PMID- 12741519 TI - Evaluation of probabilistic risk assessment of pesticides in the UK: chlorpyrifos use on top fruit. AB - The use of probabilistic risk assessment (PRA) for examining chemical impacts has become an important area of debate within the European Union. This paper describes a case study on probabilistic techniques to assess pesticide risks in the UK aquatic environment. The main aim of this paper is to demonstrate both the potential strengths and weaknesses of PRA for assessing pesticides when compared with the conventional deterministic approach, and to examine whether PRA is useful within the European regulatory context. The organophosphate insecticide, chlorpyrifos, was selected as a model compound and toxicity exposure ratios calculated using Monte Carlo analysis and different distributions of spray drift and toxicity values following application to top fruit. Chlorpyrifos is highly toxic to arthropods but less toxic to fishes. Species sensitivity followed a log normal distribution when fitted to all toxicity data. Toxicity data quantity had little influence on species sensitivity distribution model parameters when n was greater than 10 species. Below this, estimates were less accurate and precise, possibly because of the inclusion of data from many different sources. Estimates of chlorpyrifos exposure derived from the standard spray drift model differed substantially from measurements of chlorpyrifos in European surface waters. When a distribution based on measured concentrations was used in a PRA, the risk of acute fish mortality was low, and the risk of acute arthropod mortality was lower than in other scenarios, although not negligible. If PRA is used to assess pesticides, risk managers need further guidance on how to conduct a PRA and what constitutes 'unacceptable risk' under EC Directive 91/414/EEC, as judgement is required when simple trigger values are no longer used. PMID- 12741520 TI - Pesticide degradation in a 'biobed' composting substrate. AB - Pesticides play an important role in the success of modern farming and food production. However, the release of pesticides to the environment arising from non-approved use, poor practice, illegal operations or misuse is increasingly recognised as contributing to water contamination. Biobeds appear to offer a cost effective method for treating pesticide-contaminated waste. This study was performed to determine whether biobeds can degrade relatively complex pesticide mixtures when applied repeatedly. A pesticide mixture containing isoproturon, pendimethalin, chlorpyrifos, chlorothalonil, epoxiconazole and dimethoate was incubated in biomix and topsoil at concentrations to simulate pesticide disposal. Although the data suggest that interactions between pesticides are possible, the effects were of less significance in biomix than in topsoil. The same mixture was applied on three occasions at 30-day intervals. Degradation was significantly quicker in biomix than in topsoil. The rate of degradation, however, decreased with each additional treatment, possibly due to the toxicity of the pesticide mixture to the microbial community. Incubations with chlorothalonil and pendimethalin carried out in sterile and non-sterile biomix indicated that degradation, rather than irreversible adsorption to the matrix, was the main mechanism responsible for the reduction in recovered residues. Results from these experiments suggest that biobeds offer a viable means of treating pesticide waste. PMID- 12741522 TI - Adsorption-desorption of metalaxyl as affecting dissipation and leaching in soils: role of mineral and organic components. AB - Adsorption-desorption studies of metalaxyl in fifteen agricultural soil samples from Southern Portugal and Spain were performed following a batch equilibration method. Very high sorption was observed on a clay soil of high content in altered illite, but, when this soil was excluded from regression analysis, organic matter (OM) was the most important single soil property affecting sorption at low concentrations of metalaxyl. At higher concentrations, no correlation was observed with any soil property. The relevance of OM on sorption was also corroborated by the increase in sorption coefficients when soil OM was increased artificially by the addition of an organic amendment. Sorption studies with the colloidal fraction of selected soils also revealed the importance of mineral surfaces in metalaxyl sorption. Sorption of metalaxyl in most of the soil samples was hysteretic. Selected soil samples were incubated in the dark, sampled periodically and extracted for their fungicide content. Metalaxyl half-lives increased with sorption and OM content of the soil, and were specially high in the amended soil. Leaching studies in hand-packed columns under saturated/unsaturated flow conditions showed an inverse relation between leaching and sorption. Recoveries from the soil columns were close to 80% of the metalaxyl applied, except for the soil which OM was artificially increased, in which recovery was lower and this has been attributed to the much higher irreversibility of metalaxyl sorption in the amended soil. PMID- 12741521 TI - Contribution of contact toxicity and wheat condition to mortality of stored product insects exposed to spinosad. AB - Spinosad, a reduced-risk commercial insecticide derived from a bacterial fermentation product, possesses both contact and oral toxicities against insects. Contact toxicity of spinosad to adults of Rhyzopertha dominica (F), Sitophilus oryzae (L), and Tribolium castaneum (Herbst) was evaluated by exposure for 24 or 48 h to treated glass Petri dishes. Adults were exposed to different deposits (0.001-0.79 mg cm(-2)) of spinosad in 24-h tests and to deposits of 0, 0.0016 and 0.016mg cm(-2) in 48-h tests. Rhyzopertha dominica was most susceptible to spinosad in 24- and 48-h tests, followed by S. oryzae, and T. castaneum. The 24-h LD50 values were 0.0004, 0.077 and 0.189mg cm(-2) for R. dominica, S. oryzae, and T. castaneum, respectively. All R. dominica adults were dead following 48 h exposure to both spinosad deposits, whereas mortality of S. oryzae and T. castaneum ranged from 10 to 85% and 12 to 48%, respectively. Rhyzopertha dominica, T. castaneum, and O. surinamensis adults were exposed for 14 days to whole wheat, cracked wheat and wheat flour treated with 0, 0.1 and 1.0 mg kg(-1) of spinosad. Rhyzopertha dominica adults were highly susceptible to spinosad, followed by O. surinamensis and T. castaneum. Immatures (eggs and larvae) of T. castaneum and O. surinamensis exposed for 14 days were more susceptible on spinosad-treated whole wheat than on treated cracked wheat and wheat flour. This is the first report documenting contact activity of spinosad, and the effect of grain condition on spinosad toxicity, to stored-product insects. PMID- 12741523 TI - Development of a bioassay system for monitoring susceptibility in Thrips tabaci. AB - A system is described for collecting adult and larva of Thrips tabaci from onion foliage into insecticide-treated vials to evaluate susceptibility to insecticides. The thrips insecticide bioassay system (TIBS) allows one to treat vials and store them for 3 weeks before thrips are collected. Depending on the population density in the field, collection of the insects for the test required from 3-6 h for one person. Assays are read after 24h. This system was used in 1997 and 1998 in commercial onion fields in Honduras and Nicaragua, and TIBS was sensitive enough to detect differences to the insecticides tested, to thrips life stages and to different generations within an onion-growing season. Data collected suggest that there were not serious problems with thrips insecticide resistance, with the possible exception of cypermethrin in Nicaragua which had a resistance ratio (RR) value of 26.9 for adult thrips. The largest RR values were observed at the end of the growing season, and this may be caused by the season long selection by insecticide sprays. The mortality of adults and larvae followed the same general pattern, but the ratio between larvae and adults differed for each chemical group. PMID- 12741524 TI - Variations in acaricidal effect of wettable sulfur on Tetranychus urticae (Acari:Tetranychidae): effect of temperature, humidity and life stage. AB - Under laboratory conditions, the acaricidal effect of wettable sulfur is influenced by climatic conditions and the stage of development of Tetranychus urticae. Its ovicidal effect results from the combined action of temperature and relative humidity (RH). Wettable sulfur becomes effective at 27.5 degrees C and 75% RH. Beyond this threshold, the acaricidal effect increases with rising temperature or humidity, to become complete at a temperature of 35 degrees C and 90% RH. Within the range of temperatures and humidities 20 degrees C/30% RH and 35 degrees C/90% RH the mortality of immatures (from protonymphs to teleiochrysalis) was total. Under similar experimental conditions, females usually died before the end of the experiment. Temperature and relative humidity increased the adulticidal potential of wettable sulfur. The fecundity of the sulfur-treated females and the viability of their progeny was found to depend on temperature and RH. According to the same climatic conditions, eggs were less susceptible than females, which were in turn less susceptible than juvenile stages. PMID- 12741525 TI - Patch spraying: future role of electronics in limiting pesticide use. AB - Developments relating to the control of application equipment can deliver improvements in pesticide use by better matching applications to target requirements. This may have components relating to the spatial distribution of a weed, pest or disease or methods by which the target, particularly a crop canopy, can be described with respect to a given application. Changes in application can relate to the dose and/or volume applied, but may also concern the way in which a treatment is delivered in terms of parameters such as spray trajectory angle and droplet size distribution. For many weed species there is evidence of patchy distributions in field situations. Studies have shown that savings of typically up to 40% in herbicide use can be achieved by adopting patch spraying approaches in such situations. Weed patch detection is key to the performance of such patch spraying systems. In widely spaced rowcrops such as vegetables, there is considerable scope for developing fully automated detection systems based on image analysis, and for the development of accurate guidance systems that apply pesticides only to the crop row. In crops with a relatively high plant density, weed detection in the medium/short term is likely to be based on manual discrimination. The costs of labour for manual weed patch mapping have been estimated at less than 1.50 ha(-1) pounds sterling. Potential savings in pesticide use can also be made if applications are matched to crop canopy structure. This is most important in bush and tree crops where savings of up to 75% in pesticide use could be achieved. In crops such as cereals, studies have shown that savings in fungicide use may be possible, particularly at earlier stages of growth by adjusting spray delivery to measured canopy characteristics. Key components of the performance of application systems concern the ability to deliver over a dose rate range of more than 3:1 while maintaining control of variables such as delivery trajectory angle and spray quality. Traceability and the effective monitoring of applications is likely to be a major driver influencing the uptake of more sophisticated control systems. Methods of labelling pesticides with systems that can be read by the application unit will be an important step in the development of recording and data handling systems that will operate safely with the minimum of operator input and enable the environmental advantages of targeted pesticide application to be monitored. PMID- 12741526 TI - Growing vegetables in developing countries for local urban populations and export markets: problems confronting small-scale producers. AB - Vegetables attract high applications of pesticides, and farmers in developing countries use many acutely toxic insecticides to control pests on these crops. With the liberalisation of agricultural markets in developing countries, the number of small-scale farmers growing vegetables for both domestic and export markets is increasing. Demand for supplies of year-round and exotic fruit and vegetables has grown in industrialised countries, but with rising quality standards and traceability requirements it is difficult for small-scale farmers to benefit from this lucrative non-traditional agricultural export trade. The demand is high for vegetables in the expanding cities in developing countries, and farmers in peri-urban areas, or rural areas with good access to the cities, are in a position to find a growing market for their produce. Poor storage facilities will often mean that farmers are forced to sell at peak times when prices are low. Farmers rarely have access to training in pesticide use, and have only limited or no access to advice on the complicated management of pesticides. The Food and Agriculture Organisation of the UN is concerned about high levels of poor quality and adulterated pesticides on sale in developing countries. Surveys repeatedly show that without training, farmers are unable to make good crop decisions: recognition of pests and their predators is generally low, leading to decisions to spray to kill any insect; knowledge of product selection, application rates and timing is poor; different products are often combined in the belief that the effect will be greater; re-entry periods after spraying and essential harvest intervals are not known; and without knowledge of alternatives, farmers will often assume that the only solution to pest problems is to spray more frequently. From a consumer's point of view, few developing countries are able to monitor pesticide residues, particularly for produce grown for home consumption: most countries do not have laboratories for even simple residue testing. Changes in European Maximum Residue Limits means that export crops will be rejected if they contain residues at the Limit of Detection of pesticides not registered in Europe. Season-long field level training in Integrated Pest Management can help farmers to become better decision-makers, and to greatly reduce pesticide use while reducing risks to their own health and environment, producing safer products for consumers, maintaining yields, and increasing incomes. PMID- 12741527 TI - Pesticide buffer zones for the protection of wildlife. AB - Buffer zones can play important roles in agricultural habitats, both in the protection of off-crop habitats from pesticide and fertiliser drift and run-off, and in providing important areas of non-crop habitats. Their role in the protection of aquatic habitats from pesticide drift is a significant feature of pesticide risk management, but they are currently used only to a limited degree to protect terrestrial habitats. This paper summarises some of the evidence for the risks and impacts of pesticide drift into non-crop habitats, and the approaches taken by the nature conservation agencies in deriving buffer zones for the protection of sites that are important for nature conservation. Biodiversity objectives need to be explicitly built into ICM programmes, and buffer zones, together with appropriate application technology, can play an important part in reducing pesticide drift in ICM systems to achieve such objectives. PMID- 12741528 TI - Pesticide usage in some vegetable crops in Great Britain: real on-farm applications. AB - The last decade has seen an increasing requirement throughout the United Kingdom (UK) for accurate, timely information on pesticide usage on fresh produce such as vegetables to satisfy government legislation and provide data as part of the registration and review process of pesticides. These data, collected by the Department for Environment, Food and Rural Affair's Pesticide Usage Survey Group (PUSG), provide information on current and past usage patterns and allow the impact of use on the environment, operator and consumer to be more accurately assessed from actual usage patterns rather than worst-case scenarios. Surveys of commercially grown carrot, Brussels sprouts and onion crops in Great Britain were undertaken at regular intervals between 1986 and 1999, using fully stratified samples of growers. Only on Brussels sprouts was there a significant decline in pesticide use over this period, but the frequency of treatments increased on all three crops, reflecting the recent introduction of more active molecules and lower dose rates. The implications for growers of EU Directive 91/44 are discussed if active ingredients are lost because they fail to achieve Annex 1 listing. PMID- 12741529 TI - Keeping science in environmental regulations: the role of the animal scientist. AB - Environmental issues continue to be one of the biggest challenges faced by livestock producers. Whereas issues of the past have focused on manure nutrient impacts on water quality with some regulatory activity addressing odors, emerging issues are more diverse. To address emerging air quality issues, such as ammonia emissions, antibiotic transfer, human health impacts of emissions from animal agriculture, and estrogens in the environment, scientists with expertise in physiology, genetics, animal management, and nutrition will need to be enlisted. The objectives of this review are to highlight some of the prominent environmental regulatory activity that has occurred nationally in the past few years, to outline some of the emerging environmental issues, and to move members of the animal science profession toward thinking about what they can contribute toward addressing these issues. Animal scientists are uniquely qualified to engage in environmental research, education, and policymaking because of our comprehensive understanding of the complexity of whole-farm management and the interactions between animal management and manure management. Animal science departments have the opportunity to train students to be leaders in addressing environmental issues related to animal production, provided departments incorporate environmental education into curricula. Animal scientists can contribute greatly to the many areas of research that address emerging and current environmental issues, helping to ensure that policy is science-based and mitigation strategies are feasible. PMID- 12741530 TI - Soyhulls as an alternative feed for lactating dairy cows: a review. AB - Dairy producers use soyhulls, a byproduct of soybean processing, to replace either grain or forage in diets of lactating dairy cows. In view of the nutritional and economical value of soyhulls it is anticipated that this practice will continue to increase in popularity among nutritionists and producers of ruminant animals. This paper reviews information regarding the nutritional value of soyhulls and the effects of feeding this alternative feed on ruminal fermentation, nutrient digestion and utilization, and performance of dairy cows. Soyhulls can replace corn grain to supply about 30% of the dry matter (DM) in high-grain diets without negatively affecting either the fermentation or digestion of nutrients in the gastrointestinal tract or the performance of dairy cows. Additionally, data suggest that soyhulls might successfully replace forage to supply < or = 25% of the DM in diets of dairy cows when the supply of effective fiber, which includes a chemical and a physical component, remains adequate after including the hulls. However, caution should be exercised when data from different studies are extrapolated to practical situations because the response to feeding soyhulls appears to be largely affected by the type of carbohydrate being replaced by soyhulls; the amount, type, and physical form of the dietary forage; and the incidence of either negative or positive associative effects before and after the addition of soyhulls to the original diet. Unfortunately, the paucity of data from experiments in which soyhulls constituted more than 25 to 30% of the dietary DM restricts the ability to identify the maximum amount of soyhulls that can be used in diets of dairy cows. Information from studies in which > or = 25 to 30% of dietary DM supplied as either cereal grains or forages are replaced with soyhulls is needed to better understand and predict the production of dairy cows fed diets containing the hulls. This knowledge is essential for maximizing the use of soyhulls in diets for dairy cows. PMID- 12741531 TI - Low-fat set yogurt made from milk subjected to combinations of high hydrostatic pressure and thermal processing. AB - The combined use of high hydrostatic pressure (300 to 676 MPa, 5 min) and thermal treatment (85 degrees C, 30 min) in milk for the manufacture of low-fat yogurt was studied. The objective was to reduce syneresis and improve the rheological properties of yogurt, reducing the need for thickeners and stabilizers. The use of high hydrostatic pressure alone, or after thermal treatment, reduced the lightness and increased the viscosity of skim milk. However, milk recovered its initial lightness and viscosity when thermal treatment was applied after high hydrostatic pressure. The MALDI-TOF spectra of skim milk presented monomers of whey proteins after a treatment of 676 MPa for 5 min. Yogurts made from skim milk subjected to 400 to 500 MPa and thermal treatment showed increased yield stress, resistance to normal penetration, and elastic modulus, while having reduced syneresis when compared to yogurts from thermally treated or raw milks. The combined use of thermal treatment and high hydrostatic pressure assures extensive whey protein denaturation and casein micelle disruption, respectively. Although reaggregation of casein submicelles occurs during fermentation, the net effect of the combined HHP and thermal treatment is the improvement of yogurt yield stress and reduction of syneresis. PMID- 12741533 TI - Application of fluorescence spectroscopy and chemometrics in the evaluation of processed cheese during storage. AB - Front face fluorescence spectroscopy is applied for an evaluation of the stability of processed cheese during storage. Fluorescence landscapes with excitation from 240 to 360 nm and emission in the range of 275 to 475 nm were obtained from cheese samples stored in darkness and light in up to 259 d, at 5, 20 and 37 degrees C, respectively. Parallel factor (PARAFAC) analysis of the fluorescence landscapes exhibits four fluorophores present in the cheese, all related to the storage conditions. The chemometric analysis resolves the fluorescence signal into excitation and emission profiles of the pure fluorescent compounds, which are suggested to be tryptophan, vitamin A and a compound derived from oxidation. Thus, it is concluded that fluorescence spectroscopy in combination with chemometrics has a potential as a fast method for monitoring the stability of processed cheese. PMID- 12741532 TI - Influence of presalting and brine concentration on salt uptake by Ragusano cheese. AB - The impact of presalting and nonsaturated brine on salt uptake by Ragusano cheese was determined. The study included four treatments: 1) the traditional method using no presalting and saturated brine, 2) presalting and saturated brine, 3) no presalting and 18% brine for 8 d followed by 16 d in saturated brine, and 4) presalting and 18% brine for 8 d followed by 16 d in saturated brine. Cheese blocks were weighed and sampled before brine salting (time 0) and after 1, 4, 8, 16, and 24 d of brining for each treatment. Presalting delivered 60% of the normal level of salt in the center of the block prior to brine salting without decreasing the rate of uptake of salt from either saturated or 18% brine. Use of 18% salt brine for the first 8 d of 24 d of brine salting increased the rate of salt uptake, compared with 24 d in saturated brine. The increased rate of salt uptake with 18% brine compared with saturated brine was related to the impact of salt brine on the moisture content and porosity of the cheese near the surface of the block. Brine with higher salt content causes a rapid loss of moisture from cheese near the surface of the block. Moisture loss causes shrinkage of the cheese structure and decreases porosity, which impedes moisture movement out and salt movement into the block. The use of 18% salt brine for the first 8 d delayed the moisture loss and cheese shrinkage at the exterior of the block and allowed more salt penetration. PMID- 12741534 TI - Effect of frozen storage on physical properties of pasta filata and nonpasta filata Mozzarella cheeses. AB - The effects of 1) ripening 2, 7, and 14 d at 7 degrees C before freezing; 2) tempering 7, and 14 d at 7 degrees C after freezing; and 3) frozen storage for 1 and 4 wk at -20 degrees C, on the meltability, stretchability, and microstructure of pasta filata and nonpasta filata Mozzarella cheeses were investigated. Cheeses were cut into 5 x 10 x 7-cm blocks and vacuum-sealed 1 d after manufacture. The results were compared to the corresponding results obtained with unfrozen control samples, aged at 7 degrees C between 2 and 21 d. The changes in physical properties of frozen-stored pasta filata and nonpasta filata Mozzarella cheeses were consistent with critical damage to the cheese microstructure as compared to the unfrozen control samples. Generally, aging before and tempering after freezing resulted in increased meltability of both frozen-stored pasta filata and nonpasta filata Mozzarella cheeses. The stretchability of frozen-stored pasta filata Mozzarella cheese increased during tempering, but that of nonpasta filata Mozzarella cheese decreased during aging and tempering. In most cases, one-week frozen stored pasta filata Mozzarella cheese had higher meltability and stretchability than 4-wk frozen-stored sample. For 1-wk frozen-stored nonpasta filata Mozzarella cheese, the meltability increased but stretchability decreased when it was frozen-stored for 4 wk. PMID- 12741535 TI - Intracellular esterase from Lactobacillus casei LILA: nucleotide sequencing, purification, and characterization. AB - An esterase gene (estC) was isolated from a genomic library of Lactobacillus casei LILA. The estC gene consisted of a 777 bp open reading frame encoding a putative peptide of 28.9 kDa. A recombinant EstC fusion protein containing a C terminal six-histidine tag was constructed and purified to electrophoretic homogeneity. Characterization of EstC revealed that it was a serine-dependent dimeric enzyme. Optimum temperature, NaCl concentration, and pH for EstC were determined to be 30 degrees C, 0% NaCl, and pH 5.5, respectively. EstC had significant activity under conditions simulating those of ripening cheese (10 degrees C, 4% NaCl, and pH 5.1). Kinetic constants (KM and Vmax) were determined for EstC action on a variety of ethyl esters and ester compounds consisting of substituted phenyl alcohols and short n-chain fatty acids. For comparison purposes, the previously studied EstA from Lactococcus lactis MG1363 was purified to electrophoretic homogeneity and its substrate selectivity determined in a similar fashion. Different substrate selectivities were observed for EstC and EstA. PMID- 12741536 TI - Perception of melting and flavor release of ice cream containing different types and contents of fat. AB - Temporal effects of dairy and vegetable fats (0 to 18%) on perception of strawberry flavor release and melting of ice cream were studied using the time intensity sensory method. Also, aroma and flavor attributes of the ice cream samples were evaluated. Only slight effects of fat on the rate of flavor release and flavor intensity were perceived. A slightly faster flavor release from the vegetable fat compared with dairy fat was noticed. Polydextrose and maltodextrin as bodying agents in the fat-free ice cream significantly increased flavor release and melting rate of the ice cream. Increasing fat content slightly retarded melting of ice cream in the mouth. No significant effect of the fat quality on perceived melting was noticed. Significant differences in aroma and flavor attributes of the fat-free and other samples were perceived. Intensity and sharpness of the strawberry aroma and flavor were greater in fat-free samples and they were perceived as nontypical. Fattiness and creaminess were highly correlated. Maltodextrin and polydextrose increased perceived fattiness and creaminess of fat-free ice cream. PMID- 12741537 TI - High pressure effects on proteolytic and glycolytic enzymes involved in cheese manufacturing. AB - The activity of chymosin, plasmin, and Lactococcus lactis enzymes (cell envelope proteinase, intracellular peptidases, and glycolytic enzymes) were determined after 5-min exposures to pressures up to 800 MPa. Plasmin was unaffected by any pressure treatment. Chymosin activity was unaffected up to 400 MPa and decreased at 500 to 800 MPa. Fifty percent of control chymosin activity remained after the 800 MPa treatment. The lactococcal cell envelope proteinase (CEP) and intracellular peptidase activities were monitored in cell extracts of pressure treated cells. A pressure of 100 MPa increased the CEP activity, whereas 200 MPa had no effect. At 300 MPa, CEP activity was reduced, and 400 to 800 MPa inactivated the enzyme. X-Prolyl-dipeptidyl aminopeptidase was insensitive to 5 min pressure treatments of 100 to 300 MPa, but was inactivated at 400 to 800 MPa. Aminopeptidase N was unaffected by 100 and 200 MPa. However, 300 MPa significantly reduced its activity, and 400 to 800 MPa inactivated it. Aminopeptidase C activity increased with increasing pressures up to 700 MPa. High pressure did not affect aminopeptidase A activity at any level. Hydrolysis of Lys Ala-p-NA doubled after 300-MPa exposure, and was eliminated at 400 to 800 MPa. Glycolytic enzyme activities of pressure-treated cells were evaluated collectively by determining the titratable acidity as lactic acid produced by cell extracts in the presence of glucose. The titratable acidities produced by the 100 and 200 MPa samples were slightly increased compared to the control. At 300 to 800 MPa, no significant acid production was observed. These data demonstrate that high pressure causes no effect, activation, or inactivation of proteolytic and glycolytic enzymes depending on the pressure level and enzyme. Pressure treatment of cheese may alter enzymes involved in ripening, and pressure treating L. lactis may provide a means to generate attenuated starters with altered enzyme profiles. PMID- 12741538 TI - A protein kinase is located in the micellar fraction of fresh pasteurized skimmed farm milk. AB - Recombinant protein kinase CK2 from the yeast Schizosaccharomyces pombe is able to phosphorylate casein in skimmed pasteurized milk. We could incorporate up to 540 pmol of phosphate into 50 microg milk proteins, i.e., 0.26 P/mol caseins. To better understand the action of protein kinase CK2 on milk proteins, we have compared the action of rspCK2alpha on milk, and on different casein micellar subfractions isolated from milk by ultracentrifugation. In contrast to the situation observed with phosphocaseinate, alpha(s) casein was the best substrate for rspCK2alpha, whether milk or micellar fractions were used as substrates. We have characterized the protein content of different micellar fractions obtained by ultracentrifugation of cow milk using capillary zone electrophoresis. We confirm that the kappa casein content of micelles largely decreases when their size increases. In contrast, the alpha(s) casein content slightly increased with micelles size and beta casein content remained constant. All of the micellar fractions were substrates for rspCK2alpha, but a significant amount of intrinsic protein kinase activity was also found. The intrinsic protein kinase used added ATP as phosphate donor, and was only slightly sensitive to high heparin concentration. It could phosphorylate micellar casein in milk ultrafiltrate, in the absence of addition of any metallic cofactor. Its activity was only slightly affected by the addition of either MgCl2 or MnCl2. CaCl2 activated the enzyme significantly. The intrinsic kinase lost its activity with time, and could incorporate from 9 to 26% of the total phosphate incorporated in the presence of rspCK2a. Alpha(s) casein was the best substrate of the intrinsic kinase, followed by beta casein. In the presence of CaCl2, the intrinsic kinase was found to incorporate up to 470 pmol of phosphate into 50 microg of milk proteins. PMID- 12741539 TI - Properties of frozen dairy desserts processed by microfluidization of their mixes. AB - Sensory properties and rate of meltdown of nonfat (0% fat) and low-fat (2% fat) vanilla ice creams processed either by conventional valve homogenization or microfluidization of their mixes were compared with each other and to ice cream (10% fat) processed by conventional valve homogenization. Mixes for frozen dairy desserts containing 0, 2, and 10% fat were manufactured. Some of the nonfat and low-fat ice cream mixes were processed by microfluidization at 50, 100, 150, and 200 MPa, and the remaining nonfat and low-fat ice cream mixes and all of the ice cream mix were processed by conventional valve homogenization at 13.8 MPa, first stage, and 3.4 MPa, second stage. The finished frozen and hardened products were evaluated at d 1 and 45 for meltdown rate and for flavor and body and texture by preference testing. Nonfat and low-fat ice creams that usually had a slower meltdown were produced when processing their mixes by microfluidization instead of by conventional valve homogenization. Sensory scores for the ice cream were significantly higher than sensory scores for the nonfat and low-fat ice creams, but the sensory scores for the conventional valve homogenized controls for the nonfat ice cream and low-fat ice cream were not significantly different from the sensory scores for the nonfat ice cream and low-fat ice cream processed by microfluidization of the mixes, respectively. Microfluidization produced nonfat and low-fat ice creams that usually had a slower meltdown without affecting sensory properties. PMID- 12741540 TI - Characterization and proteolytic origins of specific peptides appearing during lipopolysaccharide experimental mastitis. AB - Based on the compositional change of the proteose peptone fraction, proteolysis was studied over time following lipopolysaccharide-induced experimental mastitis. Electrophoresis of the proteose peptone fraction revealed many degradation products. Five peptides were identified by amino-terminal sequencing as internal fragments of beta-, kappa-, alpha(s1)-, and alpha(s2)-casein that were generated by somatic cell proteases. Although kappa-casein is considered particularly resistant to endogenous proteolysis, a kappa-casein peptide was electrophoretically isolated in association with a beta-casein fragment. The in vitro kinetic studies of caseinate hydrolysis by elastase, one of the main polymorphonuclear neutrophil (PMN) proteases, suggested that the beta-casein peptide might be generated by elastase. In addition, elastase activity in milk PMN was higher during the inflammation of the mammary gland than prior to infusion. PMID- 12741542 TI - Optimization of DNA-based vaccination in cows using green fluorescent protein and protein A as a prelude to immunization against staphylococcal mastitis. AB - Staphylococcus aureus is a contagious pathogen that often results in chronic intramammary infections in dairy cows. Current vaccine formulations are ineffective in preventing this infection. The objective of this study was to stimulate an immune response in dairy cows through injection of plasmid DNA designed to express staphylococcal Protein A in transfected cells. Intramuscular and intradermal vaccination sites were evaluated using a plasmid containing the human cytomegalovirus (CMV) promoter/enhancer directing expression of green fluorescent protein (pcDNA3/GFP). DNA was delivered by needle and syringe, or by high-, intermediate-, or low-pressure jet injections (Ped-o-Jet and LectraJet). Five cows per treatment were injected with 0.5 mg of plasmid DNA at 6, 4, and 2 wk prepartum. Serum antibody levels determined by ELISA indicated that intradermal high-pressure jet injection elicited a greater immune response compared to needle and syringe injection. Differences in antibody production among low-pressure and needle and syringe treatment groups were not significant. An expression plasmid containing the CMV promoter/enhancer driving expression of the Fc-binding domain of S. aureus Protein A was coinjected into cows by vulvamucosal vaccination using the high-pressure Ped-o-Jet. Beginning 6 wk prepartum, groups of cows (n = 5) were injected three times at 2-wk intervals with DNA in saline, DNA in aluminum phosphate adjuvant, or served as noninjected controls. A cellular immune response to Protein A was detected in 4 of 10 animals, while cellular responses to GFP were not detected. Humoral responses to Protein A were observed in 6 of 10 animals and to GFP in 2 of 10 animals. Aluminum phosphate adjuvant appeared to enhance antibody production in response to Protein A. In experiment 3, a protein boost injection of Protein A was given to six animals approximately 5 mo postpartum. Three animals were nonvaccinated controls, and three were among those stimulated to produce antibody in response to the DNA-based vaccine. These results showed that Protein A specific antibodies remained elevated as compared to nonvaccinated controls and were stimulated in response to the protein boost. However, the magnitude of the response in animals previously vaccinated with DNA was not different than that observed in the nonvaccinated controls. We have shown that a humoral and cellular immune response to abbreviated Protein A can be raised in dairy cows using intravulvamucosal jet injection of a DNA-based vaccine. PMID- 12741541 TI - Effect of a monensin controlled release capsule on metabolic parameters in transition dairy cows. AB - A prospective field study involving 251 Holstein cows and heifers from five dairy farms near Guelph, Ontario, Canada, was conducted to measure the effect of monensin delivered in a controlled release capsule 3 wk precalving on metabolic function in dairy cows immediately pre- and postcalving. At 3 wk before expected calving, cows were randomly assigned to receive either a controlled release capsule containing monensin or to serve as negative controls. Cows were blood sampled once per week precalving and once in the week following calving, at the same time of day and the same day of the week. Serum was evaluated for beta hydroxybutyrate (BHBA), nonesterified fatty acids (NEFA), cholesterol, urea, glucose, calcium, and phosphorus. Monensin-treated cows had significantly decreased NEFA and BHBA and significantly increased concentrations of serum cholesterol and urea in the week immediately precalving. No effect of treatment was observed for calcium, phosphorus, or glucose in the precalving period. After calving, concentrations of phosphorus were lower and BHBA tended to be lower, and cholesterol and urea were higher in monensin-treated cows. There was no effect of treatment on NEFA, glucose, or calcium in the first week postcalving. Monensin treatment administered precalving significantly improved indicators of energy balance in both the immediate precalving and postcalving periods. The findings indicate better energy metabolism in monensin-treated cows as they approach calving. Improvement of energy balance before calving is important for the prevention of energy associated metabolic diseases, such as retained placenta, clinical ketosis, and displaced abomasum, which might occur immediately postcalving. PMID- 12741543 TI - Prepartum antibiotic treatment of heifers: milk production, milk quality and economic benefit. AB - Prepartum intramammary antibiotic infusion of heifer mammary glands at 7 or 14 d before expected parturition is an effective procedure for eliminating many infections in heifers during late gestation and for reducing the prevalence of mastitis in heifers during early lactation and throughout lactation. Mastitis pathogens were isolated from 76% of samples obtained from untreated control quarters 7 d before expected calving, from 47% of samples obtained 3 d after calving, and from 29% of samples obtained 10 d postpartum. Mastitis pathogens were isolated from about 30% of control quarters through 240 d of lactation. A similar percentage of samples (70%) was positive for mastitis pathogens at C-7 before antibiotic treatment. However, only 8% of samples obtained at 3 d after calving and 4% of samples obtained at 10 d postpartum from quarters of antibiotic treated heifers contained mastitis pathogens. Throughout the remainder of lactation, mastitis pathogens were isolated from an average of about 11% of quarters. The percentage of samples with mastitis pathogens was higher in untreated controls than in antibiotic-treated quarters at all sampling intervals during lactation. A similar response was observed in heifers that were treated with antibiotics at 14 d before expected parturition. Prepartum antibiotic treated heifers produced significantly more milk than control heifers and had significantly lower somatic cell count scores than untreated control heifers. These observations are likely associated with or due to the lower prevalence of mastitis pathogen isolation in prepartum antibiotic-treated heifers throughout lactation. Prepartum antibiotic-treated heifers produced 531 kg more milk than heifers in the untreated control group. Multiplying this increase by a milk price of 0.407 dollars/kg yielded a 216.24 dollars per-heifer increase in gross revenue. The cost of treatment, including the cost of testing for antibiotic residues, was estimated at 15.60 dollars for a net revenue of 200.64 dollars per heifer. Prepartum antibiotic treatment to reduce the rate of mastitis in heifers during lactation was highly effective and economically beneficial. PMID- 12741544 TI - Disruption of bovine oocytes and preimplantation embryos by urea and acidic pH. AB - Feeding cattle diets high in degradable crude protein (CP) or in excess of requirements can reduce fertility and lower uterine pH. Objectives were to determine direct effects of urea and acidic pH during oocyte maturation and embryonic development. For experiment 1, oocytes were matured in medium containing 0, 5, 7.5, or 10 mM urea (0, 14, 21, or 28 mg/dl urea nitrogen, respectively). Cleavage rate was not reduced by any concentration of urea. However, the proportion of oocytes developing to the blastocyst stage at d 8 after insemination was reduced by 7.5 mM urea. In addition, the proportion of cleaved oocytes becoming blastocysts was decreased by 5 and 7.5 mM urea. For experiment 2, putative zygotes were collected -9 h after insemination and cultured in modified Potassium Simplex Optimized Medium (KSOM). Urea did not reduce the proportion of oocytes developing to the blastocyst stage, although 10 mM urea reduced cleavage rate slightly. For experiment 3, dimethadione (DMD), a weak nonmetabolizable acid, was used to decrease culture medium pH. Putative zygotes were cultured in modified KSOM containing 0, 10, 15, or 20 mM DMD for 8 d. DMD reduced cleavage rate at 15 and 20 mM and development to the blastocyst stage at all concentrations. Results support the idea that feeding diets rich in highly degradable CP compromises fertility through direct actions of urea on the oocyte and through diet-induced alterations in uterine pH. PMID- 12741546 TI - Short communication: Net uptake of nonesterified long chain fatty acids by the perfused caudate lobe of the caprine liver. AB - The objective was to determine whether net uptake of various nonesterified long chain fatty acids differs in the caprine liver. Caudate lobes were isolated from four mature goats and perfused (1 ml/min x g wet tissue) with buffer containing 0.3 mM of each palmitic, stearic, oleic, linoleic, linolenic, eicosapentaenoic, and docosahexaenoic acids. The amount of fatty acid in the perfusate decreased over time for all fatty acids with the exception of stearic acid. There was no net uptake of stearic acid, which was significantly different from all other fatty acids examined, with the exception of oleic acid. Net hepatic uptake of oleic acid was numerically, but not significantly lower than palmitic, linoleic, linolenic, eicosapentaenoic, and docosahexaenoic acids. It was concluded that net uptake of fatty acids was similar for all fatty acids tested with the exception of stearic acid. PMID- 12741545 TI - Splanchnic metabolism of dairy cows during the transition from late gestation through early lactation. AB - Blood flow and net nutrient fluxes for portal-drained viscera (PDV) and liver (total splanchnic tissues) were measured at 19 and 9 d prepartum and at 11, 21, 33, and 83 d in milk (DIM) in 5 multiparous Holstein-Friesian cows. Cows were fed a grass silage-based gestation ration initially and a corn silage-based lactation ration peripartum and postpartum. Meals were fed at 8-h intervals and hourly (n = 8) measures of splanchnic metabolism were started before (0730 h and 0830 h) feeding at 0830 h. Dry matter intakes (DMI) at 19 and 9 d prepartum were not different. Metabolism changes measured from 19 to 9 d prepartum were lower arterial insulin and acetate, higher arterial nonesterified fatty acids and increased net liver removal of glycerol. After calving, PDV and liver blood flow and oxygen consumption more than doubled as DMI and milk yield increased, but 85 and 93% of the respective increases in PDV and liver blood flow at 83 DIM had occurred by 11 DIM. Therefore, factors additional to DMI must also contribute to increased blood flow in early lactation. Most postpartum changes in net PDV and liver metabolism could be attributed to increases in DMI and digestion or increased milk yield and tissue energy loss. Glucose release was increasingly greater than calculated requirements as DIM increased, presumably as tissue energy balance increased. Potential contributions of lactate, alanine, and glycerol to liver glucose synthesis were greatest at 11 DIM but decreased by 83 DIM. Excluding alanine, there was no evidence of an increased contribution of amino acids to liver glucose synthesis is required in early lactation. Increased net liver removal of propionate (69%), lactate (20%), alanine (8%), and glycerol (4%) can account for increased liver glucose release in transition cows from 9 d before to 11 d after calving. PMID- 12741547 TI - Potential for improving description of bovine udder health status by combined analysis of milk parameters. AB - The objective of this study was to assess the potential of a stepwise multivariate procedure to quantify cow-level udder health based on eight milk parameters: milk yield, protein percentage, fat percentage, lactose percentage, citrate percentage, somatic cell count (SCC), and two electrical conductivity parameters. The data were collected in one research herd and included 821 cow level observations. In addition to milk parameters, disease recordings and bacteriology on quarter milk samples every eighth week throughout lactation were included. A multivariate mixed model was applied to the milk parameters in a healthy subset to adjust for the following systematic factors: total mixed ration (TMR) energy density, breed-line combination, parity, stage of lactation, and season. The proportion of variance accounted for by the mixed model ranged from 0.14 to 0.82 depending on milk parameter. The adjustments estimated in the healthy subset were applied to the whole dataset, including observations pertaining to nonhealthy cows. Combined description of the adjusted variation in the milk parameters was performed with a principal component analysis. The first principal component (Prin1) described 30% of the adjusted variation and was interpreted as being the main consequences of mastitis. Finally, cluster analysis based on Prin1 separated the observations into nine clusters, which were strongly associated with udder health in terms of increasing clinical and subclinical mastitis with increasing level of Prin1. It was concluded that a multivariate approach to assess udder health from milk parameters has the potential to substantially improve description of udder health. PMID- 12741548 TI - Evaluation of the surveillance program of Streptococcus agalactiae in Danish dairy herds. AB - The aim of this study was to evaluate the Danish surveillance program of Streptococcus agalactiae in dairy herds with respect to 1) fluctuation over time of the presence of S. agalactiae in bulk tank milk, 2) sensitivity and specificity of the bacteriological method used, and 3) contamination of bulk tank milk samples with milk from other herds. From June to September 1996, bulk tank milk was sampled from 100 Danish dairy herds seven times, with intervals of 2 wk. The samples were examined for the presence of S. agalactiae by four different methods: 1) by the method approved for the program, 2) after ultrasonic treatment of the milk before examination, 3) after freezing down the milk before examination, and 4) after selective preparation of the milk. Selected strains of S. agalactiae were examined by restriction fragment length polymorphism of the gene encoding rRNA to discriminate between the isolates. Streptococcus agalactiae was found in eight of 96 herds in which S. agalactiae had never previously been found during the surveillance program. Streptococcus agalactiae was not found in all seven sampling rounds in any of the eight herds. Comparing the approved method with supplemental findings by the other methods, the estimated sensitivity was (95% confidence limits): 0.786 (0.628; 0.892) and the estimated specificity (95% confidence limits): 0.995 (0.985; 0.999). Using all four methods on the same sample could increase the sensitivity, but by comparing the methods individually, there was no significant difference between any of them (P > 0.10). In milk samples from three herds, the ribotype of S. agalactiae was the same as in milk from herds sampled just before; therefore, it could not be ruled out that cross contamination could occur. Taking into account that S. agalactiae in bulk tank milk reflects the presence of S. agalactiae in at least one udder quarter, this investigation gives further reason to assume that S. agalactiae can be seen sporadically in several herds. A surveillance program based on annual bulk tank milk sample examinations will only detect a limited number of S. agalactiae infected herds. If the overall aim is to identify herds where the infection is established, annual bulk tank milk sample examinations combined with the information of number of colonies of S. agalactiae in the sample will be sufficient. PMID- 12741549 TI - Role of dietary copper in enhancing resistance to Escherichia coli mastitis. AB - The role of dietary copper in enhancing resistance to Escherichia coli mastitis was investigated in first-lactation heifers. Twenty-three primigravid Holstein heifers were maintained on a basal (6.5 ppm copper; -Cu) diet or a diet supplemented (20 ppm) with copper sulfate (+Cu) beginning 60 d prepartum through 42 d of lactation. Liver biopsies and blood samples were taken for liver and blood minerals and plasma ceruloplasmin. Milk samples were taken weekly postpartum for bacteriology. The overall mean liver Cu concentration was about threefold higher, and the overall mean plasma Cu concentration was greater in the +Cu group than the -Cu group. At 34 d of lactation, one pathogen-free quarter per animal was infused with 22 cfu of Escherichia coli strain 727. Plasma Cu was greater at -24, 0, 18, 24, 36, 96, 192, and 240 h relative to infusion for +Cu animals. Plasma Zn concentration was higher at 24 h for the +Cu group. Milk bacterial count (log10 cfu/ml) was lower at 12, 18, and 48 h for the +Cu group. Somatic cell count (log10/ml) was lower at 18 h in +Cu animals. Clinical score at 24 h was lower for +Cu cows, while at 144 h, clinical score was lower for -Cu cows. Rectal temperature was lower at 18 h for the +Cu group. Plasma ceruloplasmin and Fe, dry matter intake and milk production did not differ. Copper supplementation reduced the clinical response during experimental E. coli mastitis, but duration was unchanged. PMID- 12741550 TI - Infusions of casein hydrolyzates into the mammary gland disrupt tight junction integrity and induce involution in cows. AB - Milk stasis triggers local stimuli, which make the tight junctions leak and trigger involution. The aim of the study was to test the hypothesis that casein hydrolyzates compromise tight junction integrity and dry-off milk secretion in dairy cows. Six repeated doses of casein hydrolyzates after each milking during 3 d caused drastic changes in mammary secretion and composition, which were associated with irreversible cessation of milk secretion. No such changes were recorded in the control glands that had been treated with nonhydrolyzed casein. Treatment with casein hydrolyzates disturbed tight junction integrity within 8 h (as indicated by changes in Na+ and K+ concentrations), reduced the concentrations of lactose precipitously, activated the plasmin activator plasminogen-plasmin system, and induced the secretion of immunoglobulin type G and lactoferrin. At the end of the 3-d treatments, we stopped milking the experimental and control glands. Milk composition 19 d later was similar in the experimental and control glands and was consistent with the composition expected in fully involuted glands. We conclude that casein hydrolyzates are among the milk-borne factors that cause the disruption of tight junction integrity and induce involution in cows. The process induced by casein hydrolyzate was more rapid and synchronized than the involution induced at drying-off. PMID- 12741551 TI - Increased nuclear factor kappaB activity in milk cells of mastitis-affected cows. AB - Bacterial mastitis is accompanied by a drastic increase in milk somatic cell count (SCC), with neutrophils being the predominant cell type found in the infected quarters. Accumulation and activation of neutrophils at the site of infection require local expression of many inflammatory genes encoding adhesion molecules, chemokines and cytokines. Most of the inflammatory genes contain binding sites for the nuclear factor kappaB (NF-kappaB) within their promoter and therefore partly depend on NF-kappaB for their expression. We thus hypothesized that an increase in NF-kappaB activity in the mammary gland could contribute to development of the neutrophilic inflammation that characterizes mastitis. In an attempt to verify this hypothesis, we first assessed milk cells from healthy and acute and chronic mastitis-affected cows for NF-kappaB activity using electrophoretic mobility shift assays. We next studied the relationships between the intensity of NF-kappaB activity in these cells and the degree of udder inflammation. Active NF-kappaB complexes were undetectable in milk cells from healthy cows, whereas high levels of NF-kappaB activity were always found in cells from cows with acute mastitis. In milk cells obtained from chronic mastitis affected cows, NF-kappaB activity varied from low to high. Finally, the level of NF-kappaB activity measured in milk cells from chronic mastitis-affected cows was not correlated to SCC or to the proportion of neutrophils present in milk samples, but was highly correlated with the expression level of interleukin-8 and granulocyte/macrophage colony-stimulating factor, two NF-kappaB-dependent cytokines crucially involved in initiation and perpetuation of neutrophilic inflammation. These results suggest that NF-kappaB might play a role in mastitis pathogenesis. PMID- 12741552 TI - Pasture forages, supplementation rate, and stocking rate effects on dairy cow performance. AB - Objectives were to evaluate effects of forage species, stocking rate, and supplementation rate on performance and physiology of grazing lactating Holstein cows under intensive rotational stocking management during summer. Eight treatments were arranged in a 2 x 2 x 2 factorial design. Animals (n = 62) grazed pastures of Tifton 85 bermudagrass or Florigraze rhizoma peanut, a tropical legume. Low and high stocking rates were 7.5 and 10.0 cows/ha for bermudagrass and 5.0 and 7.5 cows/ha for rhizoma peanut. Within each forage-stocking rate combination, cows were fed supplement at 0.33 or 0.5 kg of supplement (as-fed basis)/kg daily milk production. Cows grazing rhizoma peanut pastures produced more milk (16.9 vs. 15.4 kg/d) but had higher rectal temperatures (39.4 vs. 39.1 degrees C). Milk production per cow was improved at the higher stocking rate for bermudagrass but was reduced at the higher stocking rate for peanuts. Increasing supplementation rate boosted plasma glucose, milk production, and milk protein percent. Increased supplementation rate had a greater positive impact on milk production of cows grazing bermudagrass compared to rhizoma peanut (21.9 vs. 10.6% increase) due to a lower substitution of grain for forage intake. Organic matter intakes of forage, supplement, and total diet were greatest by cows grazing rhizoma peanut pastures and averaged 12.4, 6.1, and 18.5 kg/d compared to 9.2, 5.4, and 14.6 kg/d for cows grazing bermudagrass. Despite lower individual feed intake and performance, production per unit land area was 29% greater (112 vs. 90 kg of milk/ha per d) for cows grazing bermudagrass due to the greater stocking rate possible with that forage. Only cows supplemented at the high rate and kept at the high stocking rate on bermudagrass maintained body weight. Cows on other treatments lost body weight. Tifton 85 bermudagrass appears to be an excellent summer forage for dairy cows grazing in the southeastern U.S. given its nutritive value characteristics and high yields. Optimum stocking rate may be as high as 10 cows/ha during times of peak growth of forage for low-to-moderately producing cows fed supplement. Furthermore, the positive milk production response to additional supplement when cows grazed Tifton 85 pastures (0.8 kg/kg of supplement), indicates the value of providing supplement to cows grazing this moderate quality forage. PMID- 12741553 TI - Purine derivative excretion in dairy cows: endogenous excretion and the effect of exogenous nucleic acid supply. AB - An experiment was conducted with dairy cows to study the partitioning of excreted purine derivatives between urine and milk and to quantify the endogenous contribution following the isotopic labeling of microbial purine bases. Three lactating cows in their second lactation that had been cannulated in the rumen and the duodenum were fed a mixed diet (48:52, roughage/concentrate ratio) distributed in equal fractions every 2 h, and duodenal flow of purine bases was determined by the dual-phase marker system. Nitrogen-15 was infused continuously into the rumen to label microbial purine bases, and the endogenous fraction was determined from the isotopic dilution in urinary purine derivatives. Urinary and milk recovery of duodenal purine bases were estimated at early (wk 10) and late (wk 33) lactation by the duodenal infusion of incremental doses (75 and 150 mmol purine bases/d) of RNA from Torula yeast. Each period was 6 d, with RNA being infused during the last 4 d, followed by measurement of the flow of purine bases to the duodenum. The isotope dilution of purine derivatives in urine samples confirmed the presence of an endogenous fraction (512 +/- 36.43 micromol/W0.75 or 56.86 mmol/d) amounting to 26 +/- 3.8% of total renal excretion. Total excretion of purine derivatives in urine plus milk was linearly related to the duodenal input of purine bases, but the slopes differed (P < 0.005) between lactation stages resulting in a lower equimolar recovery in early (y = 58.86 (+/-3.89) +0.56 (+/-0.0164) x; r = 0.90) than late lactation (y = 58.86 (+/-3.89) + 0.70 (+/-0.046) x; r = 0.80). Excretion of purine derivatives through milk represented a minimum fraction of total excretion but responded significantly to the duodenal input of purine bases. No differences between lactation stages were detected, and variations in milk yield did modify significantly the amount of purine derivatives excreted through the milk. PMID- 12741554 TI - Effect of feeding protein supplements of differing degradability on omasal flow of microbial and undegraded protein. AB - Ten ruminally cannulated lactating Holstein cows that were part of a larger trial studying the effects of feeding different proteins on milk production were used in a replicated 5 x 5 Latin square to quantify flows of microbial and rumen undegradable protein (RUP) in omasal digesta. Cows were fed total mixed rations containing (dry matter basis) 44% corn silage, 22% alfalfa silage, 2% urea, and 31% concentrate. The basal diet contained 31% high-moisture corn; equal N from one of four protein supplements was added to the other diets at the expense of corn: 9% solvent soybean meal (SSBM), 10% expeller soybean meal (ESBM), 5.5% blood meal (BM), and 7% corn gluten meal (CGM). Omasal sampling was used to quantify total AA N (TAAN) and nonammonia N (NAN) flows from the rumen. Estimates of RUP were made from differences between total and microbial N flows, including a correction for RUP in the basal diet. Modifying a spectrophotometric assay improved total purine recovery from isolated bacteria and omasal samples and gave estimates of microbial TAAN and NAN flows that were similar to a standard HPLC method. Linear programming, based on AA patterns of the diet and isolated omasal bacteria and ruminal protozoa, appeared to overestimate microbial TAAN and NAN flows compared to the purine assays. Yields of microbial TAAN and NAN determined using any method was not affected by diet and averaged 32 to 35 g NAN per kilogram of organic matter truly digested in the rumen. On average, National Research Council (NRC) equations underpredicted microbial N flows by 152 g/d (vs. HPLC), 168 g/d (vs. spectrophotometry), and 244 g/d (vs. linear programming). Estimates of RUP (means from the HPLC and spectrophotometric methods) were: SSBM, 27%, ESBM, 45%, BM, 60%, and CGM, 73%. Except for CGM, RUP values averaged about 20 percentage units lower than those reported by the NRC. PMID- 12741555 TI - Responses of primiparous and multiparous Holstein cows to additional energy from fat or concentrate during summer. AB - Supplemental fat has been advocated for use during hot weather and often increases milk yield of cows past peak production when energy intake should not be limiting. Relative responses of primiparous and multiparous cows to supplemental fat or isocaloric addition of concentrates under hot weather conditions have not been determined. Nine multiparous and nine primiparous Holstein cows (154 and 167 d in milk, respectively) were used in a replicated 3 x 3 Latin square design with 28-d periods. Diets were 1) control (35% alfalfa silage, 25% corn silage, and 40% concentrate, dry matter [DM] basis); 2) control plus 3% fat (HF); and 3) high concentrate ([HC] 15% alfalfa silage, 25% corn silage, and 60% concentrate). Diets were isonitrogenous; diets HF and HC were isocaloric (1.60 Mcal of net energy for lactation [NE(L)] per kilogram DM) and higher energy than the control (1.52 Mcal/kg). No parity x diet interactions approached significance. DM intake (DMI) was greater when cows were fed HC than when they were fed HF (21.0, 20.1, and 21.3 kg/d for control, HF, and HC, respectively); intake of NE(L) tended to be increased only for HC. Milk yield was increased by higher-energy diets, but milk fat content was decreased. Milk total protein content was decreased by HF and increased by HC. Yield of solids corrected milk (SCM) was not different among diets. Efficiency of milk production, expressed either as total milk solids yield per kilogram of DMI or as kilograms of SCM per megacalorie of NE(L) intake, was greater for HF than for HC. Plasma glucose was higher after feeding for cows fed HC; plasma nonesterified fatty acids were greater for HF. Respiration rate and rectal temperature were greater for HC than for HF. Regardless of parity, increased energy density from either fat or concentrate increased milk yield in midlactation cows, but diets caused energy to be partitioned differently among milk components and body storage. Supplemental rumen-active fat had modest advantages over additional starch-based concentrate during summer heat conditions. PMID- 12741556 TI - Concentrations of sialyloligosaccharides in bovine colostrum and milk during the prepartum and early lactation. AB - Sialyloligosaccharides and sialylglycoconjugates in colostrum and milk are regarded to be important biological components with respect to be source of brain gangliosides in infant and to be antiinfectional components for the attack by the pathogenic bacteria and virus. Several acidic oligosaccharides have been characterised in both bovine and human milk or colostrum. The sialyloligosaccharide content of human colostrum and milk has been extensively studied, whereas that of cows milk and colostrum has received less attention. In this study, the concentrations of three sialyloligosaccharides of bovine colostrum and milk were determined at various stages during the prepartum and the first 7 d postpartum. The concentration of 3'SL (Neu5Ac(alpha2-3)Gal(beta1-4)Glc) reached a maximum value of 0.85 mg/ml immediately following parturition while the concentrations of 6'SL (Neu5Ac(alpha2-6)Gal(beta1-4)Glc) and 6'SLN (Neu5Ac(alpha2 6)Gal(beta1-4)GlcNAc) of 0.14 and 0.12 mg/ml, respectively, were much lower at this initial stage, although these concentration were maximum immediately following parturition. Bovine colostrum, especially that collected immediately after parturition, may be suitable as a source of 3'SL and other sialyloligosaccharides for use as additives by the food or pharmaceutical industries. PMID- 12741557 TI - Short communication: Postruminal infusion of conjugated linoleic acids negatively impacts milk synthesis in Holstein cows. AB - In view of the potential of rumen-protected conjugated linoleic acid (CLA) as a means to increase the CLA content of bovine milk, a study was undertaken to evaluate the effect of synthetic CLA on milk production and composition. Four Holstein cows received abomasal infusion of: 1) control, no lipid infusion, 2) 150 g/d of synthetic CLA, 31.7% cis-9, trans-11; 30.4% trans-10, cis-12, 3) 150 g/d of safflower oil, and 4) 150 g/d of tallow. Infusion was carried out for 20 to 22 h/d for 11-d periods in a 4 x 4 Latin square design. The milk fat concentration of cis-9, trans-11 and trans-10, cis-12 isomers of CLA was significantly increased with infusion of CLA. However, CLA infusion had other unexpected effects on milk production and composition. Milk yield dropped significantly during the period of CLA infusion. Furthermore, as well as the typical depression in milk fat reported with trans-10 isomers of CLA, other negative effects specific to CLA infusion were observed including a drop in lactose concentration and yield, a drop in protein yield, and an elevated somatic cell count. The important difference between synthetically produced CLA and CLA produced naturally in the cow is the much higher proportion of trans-10 isomers of CLA in the former. The results of this study suggest that the extent of enrichment possible for trans-10 isomers of CLA, and hence the usefulness of synthetic CLA for this purpose, may be limited because of unacceptable effects on milk yield and composition. PMID- 12741558 TI - Stabilization of returned dairy products by ensiling with straw and molasses for animal feeding. AB - Returned dairy products which are transferred to landfills might add to the environmental pollution. Such products have a high nutritional value for ruminants, but they should be stabilized to enable their use as cattle feed. The purpose of the current study was to examine stabilization of returned dairy products by ensiling in combinations with straw and molasses for animal feeding. Treatments included combinations of milk and cottage cheese with straw and molasses. Results indicate that such products ensile well with straw, and after 3 d of ensiling the pH decreased to around 4.0. It was necessary to supplement cottage cheese with molasses, to supply a carbohydrate source for the lactic acid fermentation. The major fermentation product was lactic acid. Percentage of ammonia N (of total N) was generally higher in the silages made with cottage cheese than in those made with milk; the highest percentage (16%) was measured in the second experiment in the silages prepared with cottage cheese and straw. The study indicates the potential of stabilizing returned dairy products for animal feeding along with straw and molasses. There may also be potential for large dairy farms, or groups of smaller farms, to ensile waste milk with straw for later use as feed. PMID- 12741559 TI - Pasture intake and substitution rate effects on nutrient digestion and nitrogen metabolism during continuous culture fermentation. AB - A continuous culture system was used to investigate ruminal digestion in response to increased pasture intake and three different substitution rates (SR) in a 4 x 4 Latin square design. The treatments were 1) low pasture (55 g dry matter (DM)/d, 2) medium pasture (MP, 65 g DM/d), 3) high pasture (75 g DM/d), and 4) pasture (45 g DM/d) plus concentrate (PC, 30 g DM/d). Treatments were designed to produce a low (0.33), medium (0.67), and high (1.00) SR (g of pasture/g of concentrate) by contrasting the low, medium, and high pasture intake treatments with the pasture plus concentrate treatment, respectively. Pasture was fed at 0630, 1000, 1730, and 2100 h, and concentrate at 0600 and 1700 h. Digestibility of DM and neutral detergent fiber were not affected by the amount of pasture. As the amount of pasture increased, pH decreased linearly, and total volatile fatty acid and NH3-N concentrations, and nonammonia N and bacterial N flows increased linearly. Concentrate supplementation did not affect DM digestibility at high SR but increased DM digestibility at low SR. Concentrate supplementation reduced pH and NH3-N concentrations at the three SR. Concentrate supplementation reduced the ratio of rumen degradable N to rumen degradable organic matter; however, the mechanism depended on the SR. High SR, concentrate supplementation reduced rumen degradable N, which reduced NIH-N concentration without affecting bacterial N flow. At low SR, concentrate supplementation increased rumen degradable organic matter, which reduced NH3-N concentration and increased bacterial N flow. Based on these results, at low SR, concentrate supplementation may enhance animal performance because of higher total DM intake and synthesis of microbial protein. PMID- 12741560 TI - Effects of increasing levels of refined cornstarch in the diet of lactating dairy cows on performance and ruminal pH. AB - Our study investigated the effect of a linear increase in level of ruminally fermentable carbohydrate, at a constant level of dietary starch and fiber, on performance, microbial N yield, chewing activity, and ruminal pH of midlactation dairy cows. Eight cows (53 DIM) were assigned to four treatments in a double 4 x 4 Latin square. Diets consisted of increasing levels of refined cornstarch (0, 5.9, 11.9, and 17.9% of diet dry matter) replacing dry cracked, shelled corn so that increasing amounts of dietary starch originated from refined cornstarch. Corn gluten feed was used to balance diets for similar NDF content. The four diets averaged 17.9% CP, 27.2% NDF, 18.7% ADF, and 31.1% starch (dry matter basis). Diets were fed for ad libitum intake and had a forage to concentrate ratio of 40:60. Forage was coarsely chopped (13.7 mm mean particle size) alfalfa silage. Daily dry matter intake averaged 26.0 kg and tended (P = 0.08) to increase quadratically with increasing level of refined cornstarch. Milk production averaged 38.9 kg/d and milk fat percentage tended (P = 0.08) to decrease linearly, whereas percentage of protein increased quadratically, with increasing level of refined cornstarch. Yield of components and energy corrected milk was similar across diets. Total tract digestibility of starch increased linearly from 85.1% to 92.4% with increasing level of refined cornstarch. Microbial yield was unaffected by diet and averaged 371.1 g N/d. Time spent eating decreased linearly from 329 to 308 min/d when level of refined cornstarch was increased, but rumination time was unaffected. Ruminal concentration and proportion of acetate decreased linearly while concentration and proportion of propionate increased linearly with increasing level of refined cornstarch. Mean ruminal pH, time spent below pH 5.8 (h), and area below pH 5.8 (h x pH units/d) were unaffected by level of refined cornstarch and averaged 5.97, 8.4, and 2.9, respectively. Increasing the level of carbohydrates fermented in the rumen by replacing dry cracked corn with refined cornstarch (up to 57% of dietary starch) did not compromise rumen fermentation or affect performance of midlactation dairy cows. PMID- 12741561 TI - Reduced fatty acid synthesis and desaturation due to exogenous trans10,cis12-CLA in cows fed oleic or linoleic oil. AB - To determine effects of an elevated supply of cis9,trans11-18:2 (9/11CLA) or trans10,cis12-18:2 (10/12CLA) on de novo synthesis and desaturation of long-chain fatty acids, four Holstein cows fed high-oleic sunflower (OLE) or high-linoleic safflower oil (LIN) at 2.5% of DM were infused (0.625 g/h) with 9/11CLA or 10/12CLA for 48 h via the abomasum. Treatments were assigned in a 2 x 2 factorial design. The assigned diets were fed for 11 d before each 48-h infusion period. Milk samples were obtained at 12 and 0 h before infusion and at 12-h intervals from 0 to 96 h. Concentrations of trans11-18:1 and 18:2n-6 in arterial plasma phospholipid, triglyceride, and FFA fractions were greater due to feeding LIN compared with OLE. Infused 9/11CLA and 10/12CLA were incorporated into plasma triglycerides and FFA primarily. Exogenous 10/12CLA also was found in plasma phospholipids. Milk yield and DMI were not affected by treatments. Percentages and yields of protein, lactose, and SNF in milk also were not affected by treatments. Milk fat percentage and yield, however, decreased 25% from 0 to 96 h in response to infusion of 10/12CLA compared with 9/11CLA. Yields of trans11 18:1, 9/11CLA and 18:2n-6 in milk fat before infusion were higher when LIN was fed compared with OLE. Infusion of 9/11CLA, regardless of diet, increased 9/11CLA in milk fat by 44%. Although 10/12CLA was not detectable in milk fat before infusion, it averaged 6 mg/g of total fatty acids and 2 g/d after 48 h. At 48 h, recovery in milk of infused 9/11CLA was 16% compared with 8% for 10/12CLA. Yields of saturated 6:0 to 16:0, cis9-18:1, 9/11CLA, and 20:4n-6 were reduced by 10/12CLA infusion. Due to a 40% increase in the concentration of 18:0 by 48 h of 10/12CLA infusion, however, yield of 18:0 was not affected. Ratios of cis9 18:1/18:0, 9/11CLA/trans11-18:1, and 20:4n-6/18:2n-6 in milk fat decreased in response to infusion of 10/12CLA, regardless of diet. At peak concentration of 10/12CLA, reductions in cis9-18:1 and saturated 4:0-16:0 yields accounted for 36% and 53% of the decrease in total fatty acid yield. Results indicated 10/12CLA alters lipid metabolism in the bovine mammary gland by simultaneously reducing de novo synthesis and desaturation. Furthermore, milk triglyceride synthesis may have a stringent requirement for endogenously synthesized oleic acid. PMID- 12741562 TI - Effects of varying dietary protein and energy levels on the production of lactating dairy cows. AB - Forty-five multiparous and 18 primiparous Holstein cows were fed three levels of crude protein (CP), each at three levels of neutral detergent fiber (NDF), to identify optimal dietary CP and energy. Cows were blocked by parity and days in milk into seven groups of nine and randomly assigned to an incomplete 9 x 9 Latin square trial with four, 4-wk periods. Diets were formulated from alfalfa and corn silages, high-moisture corn, soybean meal, minerals, and vitamins. Forage was 60% alfalfa and 40% corn silage on all diets; NDF contents of 36, 32, and 28% were obtained by feeding 75, 63, and 50% forage, respectively. Dietary CP contents of 15.1, 16.7, and 18.4% were obtained by replacing high-moisture corn with soybean meal. Production data were from the last 2 wk of each period. Spot fecal and urine samples were collected from 36 cows to estimate N excretion using fecal indigestible acid detergent fiber (ADF) and urinary creatinine as markers. There were no interactions (P > or = 0.08) between dietary CP and NDF for any trait; thus, effects of CP were not confounded by NDF or vice versa. Intake of DM and fat yield were lower on 15.1% CP than at higher CP. There were linear increases in milk urea and urinary N excretion and linear decreases in N efficiency with increasing CP. Increasing CP from 15.1 to 18.4% reduced milk N from 31 to 25% of dietary N, increased urinary N from 23 to 35% of dietary N, and reduced fecal N from 45 to 41% of dietary N. Decreasing NDF gave linear increases in BW gain, yield of milk, protein, true protein, lactose, and SNF, and milk/DM intake and milk N/N intake, and linear decreases in milk urea. However, fat yield was lower on 28% than 32% NDF. Reducing NDF from 36 to 28% increased purine derivative excretion by 19%, suggesting increased microbial protein. Increasing CP by adding soybean meal to diets fed cows averaging 34 kg/d of milk increased intake and fat yield but depressed N efficiency. Increasing dietary energy by reducing forage improved milk yield and efficiency and decreased excretion of environmentally labile urinary N. PMID- 12741563 TI - Effects of forage particle size, forage source, and grain fermentability on performance and ruminal pH in midlactation cows. AB - Our study investigated the effects of, and interactions between, forage particle size, level of dietary ruminally fermentable carbohydrate (RFC), and level of dietary starch on performance, chewing activity, and ruminal pH for dairy cows fed one level of dietary NDF. Twelve cows (48 DIM) were assigned to six treatments in a replicated 6 x 6 Latin square. Treatments were arranged in an incomplete 2 x 2 x 2 factorial design. Factors were: dry cracked shelled corn (DC, low RFC) or ground high-moisture corn (HMC; high RFC), finely chopped or coarse silage, and alfalfa silage as the only forage or a 50:50 ratio (DM basis) of alfalfa and corn silage. Diets combining HMC with only alfalfa silage were not included in the experiment. Diets were fed for ad libitum intake as a TMR with a concentrate:forage ratio of 61:39. Diets based on only alfalfa silage and diets based on a mix of alfalfa and corn silage averaged 18.6 and 15.8% CP, 25.8 and 24.7% NDF, 17.7 and 14.8% ADF, and 29.1 and 37.3% starch, respectively. Mean particle sizes were 5.3, 2.7, 5.6, and 2.8 mm for coarse alfalfa, fine alfalfa, coarse corn silage, and fine corn silage, respectively. Decreasing forage particle size decreased DMI (23.3 vs. 21.6 kg) and organic matter intake (22.0 vs. 20.2 kg). Increasing RFC decreased DMI (22.8 vs. 21.0 kg) and organic matter intake (21.5 vs. 20.0 kg). Decreasing forage particle size increased energy corrected milk for alfalfa based diets (34.9 vs. 37.4 kg). Percentage of milk fat decreased with decreasing forage particle size (3.07 vs. 2.90%) and increased level of RFC (3.04 vs. 2.57%). Percentage of protein increased when corn silage partially replaced alfalfa silage (2.84 vs. 2.90%) but decreased when HMC replaced DC (2.90 vs. 2.84%). Apparent total tract digestibility of DM (66.7 vs. 68.5%), OM (65.9 vs. 70.7%), and starch (88.9 vs. 93.4%) increased when level of RFC was increased. Increasing level of RFC decreased mean ruminal pH from 5.82 to 5.67 and decreased minimum pH. Hours per day at which pH was <5.8, and area <5.8, increased when corn silage partially replaced alfalfa silage (2.6 vs. 4.4 h and 8.9 h x pH vs. 11.4 h x pH) and decreased further when level of RFC was increased (4.4 vs. 6.4 h and 11.4 h x pH vs. 14.3 h x pH). Decreasing forage particle size in HMC diets increased hours and area <5.8, but for DC diets, the effect of forage particle size depended on forage source. Interactions were found between level of physically effective fiber, forage source, and level of RFC on production and pH, complicating the inclusion of these effects in dairy ration formulation and evaluation. PMID- 12741564 TI - Effects of intraruminal infusion of sodium, potassium, and ammonium on hypophagia from propionate in lactating dairy cows. AB - The objective of this experiment was to evaluate effects of salt type on hypophagic effects of intraruminal infusion of propionate in lactating dairy cows. Our working hypothesis is that oxidative metabolism of propionate causes satiety by increasing hepatic ATP concentration and decreasing the discharge rate of the hepatic vagus. We hypothesized that hypophagic effects of propionate are reduced by ammonium and potassium. We speculated that ammonium infusion lowers hepatic ATP concentration because ATP is used for urea synthesis and potassium increases the discharge rate of the hepatic vagus. Eight ruminally cannulated Holstein cows in midlactation were used in a duplicated 4 x 4 Latin square design experiment. Treatments were intraruminal infusion of propionic acid, ammonium propionate, sodium propionate, and potassium propionate. Treatment solutions were 0.93 M for propionate and 0.67 M for salts among the treatments except for propionic acid. Treatment solutions were infused over 14 h starting 2 h before feeding at 17.9 ml/min, which is equivalent to 16.7 and 11.9 mmol/min for propionate and salts, respectively. Infusion of ammonium propionate decreased dry matter intake compared with sodium propionate and potassium propionate (P < 0.04; 11.0 vs. 14.0 and 13.9 kg/12 h) by decreasing meal frequency without affecting meal size, indicating that ammonium delayed the sense of hunger. No difference in DMI and feeding behavior was observed between infusion of sodium and potassium propionate. Contrary to the hypothesis, ammonium infusion did not reduce hypophagic effects of propionate, possibly because the urea cycle indirectly stimulated oxidative metabolism in the liver by generating oxidizable carbon from amino acid catabolism. PMID- 12741565 TI - Effect of dietary cobalt supplementation on cobalt metabolism and performance of dairy cattle. AB - Three studies were conducted with dairy cattle fed diets with added Co. The first study examined cow age and added dietary Co on Co in liver and blood. Nonpregnant, nonlactating Holstein cows were blocked by age (2.5 or 6.5 yr) and assigned to either a control diet or a diet supplemented with 9 mg Co per day. The Co concentration of liver, taken on d 60, was not affected by dietary Co but was higher in the younger cows. The cytosolic fraction of liver contained the most Co, and the subcellular distribution of Co was not affected by total Co in liver. In a second study, Holstein cows were assigned to one of three treatments of dietary Co from 21 d prepartum until 120 d postpartum. There was an interaction of time x treatment x parity such that milk yield response to Co supplementation differed between multiparous cows and primiparous cows. Supplemental Co did not increase Co in serum, colostrum, milk, or liver. Primiparous cows secreted colostrum and milk with higher Co concentrations than did multiparous cows. Likewise, serum B12 levels were higher in primiparous than multiparous cows and declined with increasing days in milk (DIM). Serum Co also decreased from 7 to 120 DIM. In a final study, a Co supplement in the starter diet did not affect Co in serum or liver of young calves. In conclusion, supplemental dietary Co did not affect secretion of Co in milk, tissue retention, or subcellular distribution of Co within the liver. Primiparous and multiparous cows differed in their milk yield response to dietary Co supplementation. PMID- 12741566 TI - Evaluation of different energy feeding systems with production data from lactating dairy cows offered grass silage-based diets. AB - A set of data from 838 lactating dairy cows, drawn from 12 long-term feeding studies (at least 8 wk/period), was used to evaluate the energy feeding systems for dairy cows currently adopted in Australia, France, The Netherlands, the United Kingdom, and the United States. The animals were offered mixed diets of concentrates, forage [grass silages (n = 33) and corn silages (n = 5)] ad libitum. Data used in the present evaluation were either measured [dry matter (DM) intake, milk production and live weight], measured/estimated [dietary metabolizable energy (ME) concentration] or estimated [milk energy output and live weight change (LWC)]. The mean-square prediction error (MSPE) was used for the evaluation. Total ME intake, milk yields, and LWC varied from 91 to 338 MJ/d, 7.7 to 48.9, and -1.23 to 1.73 kg/d, respectively. Australian and French systems predicted total energy requirement and milk yield relatively well, while British, Dutch and American systems underpredicted total energy requirement by proportionately 0.06, 0.04, and 0.03, respectively; and overpredicted milk yield by 0.09, 0.06, and 0.04. The Agricultural and Food Research Council (AFRC) each produced a relatively larger error of the bias (predicted - actual data) over the total MSPE for ME requirement and milk yield and a relatively smaller error of random than other systems. However, an addition of proportionately 0.05 to the total predicted ME requirement of AFRC, as suggested in this system and currently used in the UK, indicated the prediction accuracy of ME requirement and milk yield is similar to Australian and French systems. Nevertheless, all the systems had a poor prediction of LWC. For each system, the total prediction error (total MSPE) was mainly derived from the line (slope; 0.49 to 0.64 of total MSPE), while less derived from the random (0.20 to 0.48 of total MSPE), indicating a large variation between the predicted and actual LWC existed among individual cows. The residual plots of the residual differences in LWC against predicted LWC revealed that the prediction error was greater with increasing LWC. It is concluded that Australian and French systems have a better prediction of total energy requirement and milk yield than other systems, and LWC is an inappropriate indicator of energy balance in lactating dairy cows. PMID- 12741567 TI - Kinetics of in sacco fiber-attachment of representative ruminal cellulolytic bacteria monitored by competitive PCR. AB - Stems of orchardgrass hay in nylon bags were incubated in the rumens of three ruminally fistulated sheep to monitor the rate and extent of fiber attachment by the representative ruminal cellulolytic bacteria via competitive polymerase chain reaction. After incubation for 5 min, the numbers of Fibrobacter succinogenes and the two ruminococcal species attached to stems were 10(5) and 10(4)/g dry matter (DM) of stem, respectively. At 10 min, the numbers of all three species attached to stems increased 10-fold. Thereafter, attached cell numbers of the three species gradually increased and peaked at 24 h (10(9)/g DM for F. succinogenes and 10(7)/g DM for Ruminococcus flavefaciens) or 48 h (10(6)/g DM for Ruminococcus albus). On the other hand, cell numbers of all three species in the whole digesta were constant over 24 h. Changes in the rate of in sacco neutral detergent fiber disappearance of hay stem, which showed a linear increase up to 96 h, were not synchronized with changes in cellulolytic bacterial mass. These results suggest that sufficient numbers of cells of the three cellulolytic species to move to new plant fragments are present at the start of incubation, the initial attachment to new plant matter is mostly accomplished within 10 min and then bacterial growth and fibrolytic action follow. F. succinogenes was most dominant, both in the whole rumen digesta and on the suspended hay stems, demonstrating the ecological and functional significance of this species in ruminal fiber digestion. PMID- 12741568 TI - Effects of changes in dietary amino acid balance on milk yield and mammary function in dairy cows. AB - Two experiments were conducted to determine whether longer-term deficiencies in the supply of limiting amino acids would be accompanied by a decline in mammary function (total DNA, cell proliferation rate and activities of key enzymes), and whether this would adversely affect the cow's ability to respond to a return to a nutritionally adequate diet. The first experiment was performed in early/mid lactation, and the second, using the same cows, was carried out in mid/late lactation. A control group of six cows were given a grass silage-cereal diet containing fish meal as the sole protein supplement (amino acid adequate) throughout the experiments, whereas another group of six cows in treatment received the control diet for 2 wk (lactation wk 5 and 6) and then were changed to a diet in which the fish meal was replaced by an equivalent amount of protein as feather meal (amino acid deficient) for 6 wk before returning to the fish meal diet for 4 wk (Experiment 1). After a rest period of 5 wk, the experimental procedure was repeated (Experiment 2). Although there was a fall in milk yield as lactation advanced, leading to lower milk yields in Experiment 2, the marked difference in milk yield between treatments was similar for the two stages of lactation (21% vs 16% in Experiment 1 and 2, respectively). In both experiments, the marked fall of milk yield in cows given the feather meal diet was completely recovered by a return to the fish meal diet. Despite the markedly lower milk yield with the amino acid-deficient diet, however, there was no clear evidence of corresponding changes in measurements of mammary function. PMID- 12741569 TI - The effect of reducing alfalfa haylage particle size on cows in early lactation. AB - The objective of this experiment was to evaluate effects of reducing forage particle size on cows in early lactation based on measurements of the Penn State Particle Separator (PSPS). Eight cannulated, multiparous cows averaging 19 +/- 4 d in milk and 642 +/- 45 kg BW were assigned to one of two 4 x 4 Latin Squares. During each of the 23-d periods, animals were offered one of four diets, which were chemically identical but included alfalfa haylage of different particle size; short (SH), mostly short (MSH), mostly long (MLG), and long (LG). Physically effective neutral detergent fiber (peNDF) was determined by measuring the amount of neutral detergent fiber retained on a 1.18 mm screen and was similar across diets (25.7, 26.2, 26.4, 26.7%) but the amount of particles >19.0 mm significantly decreased with decreasing particle size. Reducing haylage particle size increased dry matter intake linearly (23.3, 22.0, 20.9, 20.8 kg for SH, MSH, MLG, LG, respectively). Milk production and percentage fat did not differ across treatments averaging 35.5 +/- 0.68 kg milk and 3.32 +/- 0.67% fat, while a quadratic effect was observed for percent milk protein, with lowest values being observed for LG. A quadratic effect was observed for mean rumen pH (6.04, 6.15, 6.13, 6.09), while A:P ratio decreased linearly (2.75, 2.86, 2.88, 2.92) with decreasing particle size. Total time ruminating increased quadratically (467, 498, 486, 468 min/d), while time eating decreased linearly (262, 253, 298, 287 min/d) with decreasing particle size. Both eating and ruminating per unit of neutral detergent fiber intake decreased with reducing particle size (35.8, 36.7, 44.9, 45.6 min/kg; 19.9, 23.6, 23.5, 23.5 min/kg). Although chewing activity was closely related to forage particle size, effects on rumen pH were small, indicating factors other than particle size are critical in regulating pH when ration neutral detergent fiber met recommended levels. Feeding alfalfa haylage based rations of reduced particle size resulted in animals consuming more feed but did not affect milk production. PMID- 12741570 TI - Supplemental lactoferrin improves health and growth of Holstein calves during the preweaning phase. AB - Lactoferrin is a milk protein that exhibits broad-spectrum antimicrobial properties. Previous studies indicated that supplemental lactoferrin may alter the microbial populations in the gut of nonruminants and increase preweaning weight gains in calves. In the present study, 40 Holstein calves were used to examine the effects of supplemental lactoferrin (0, 1, 2, or 3 g/d) on health, growth, and feed intake from 3 d of age to 2 wk postweaning. Lactoferrin was mixed and fed with a nonmedicated milk replacer. Calves were housed in individual pens and offered a textured, nonmedicated starter and water for ad libitum consumption. Body weight and heart girth were measuredweekly. Intakes of milk replacer and starter were determined daily. Fecal consistency was monitored three times per week. Calves were weaned when they met certain criteria based on body weight gain and starter intake. Preweaning fecal score responded quadratically, with the group fed 1 g/d of lactoferrin having the lowest score. Overall and preweaning number of days medicated responded in the same manner as fecal score. Preweaning average daily gain and gain-to-feed ratio increased linearly with lactoferrin supplementation, whereas postweaning gain-to-feed ratio decreased linearly with lactoferrin. Overall average daily heart girth gain increased linearly with lactoferrin. Body weight, weaning age, and dry matter intake were not different among treatments. Based on the observed improved gain-to-feed ratios, increased average daily gains, improved fecal scores, and reduced morbidity in preweaned calves, it appears that lactoferrin may be a beneficial supplement in the diets of neonatal calves prior to weaning. PMID- 12741571 TI - Feed intake, milk yield, and metabolic parameters prior to left displaced abomasum in dairy cows. AB - As left-displaced abomasum (LDA) often occurs in cows with high contents of fat in the liver (fatty liver), a postpartum fatty liver-inducing regimen was applied to 16 cows. The main interest of the study was whether there were productive or metabolic changes in cows prior to LDA. Therefore, feed intake and milk production were monitored and blood samples were collected from the cows. The LDA occurred in 4 out of 16 dairy cows that were included in the feeding regimen. Compared to cows not developing LDA, LDA-cows had a significantly lower feed intake, 6.5 kg/d less, and milk production, 8 kg/d less, prior to clinical diagnosis of LDA. In the 10-d period preceding clinical diagnosis of LDA, blood concentrations of calcium, glucose, and insulin were significantly lower, whereas blood concentrations of nonesterified fatty acids and beta-hydroxybutyrate, as well as aspartate aminotransferase activities were significantly elevated compared to cows not developing LDA. These preclinical changes may play an important role in the pathogenesis of LDA. It is not certain, however, whether there is a causal association between these parameters and LDA. PMID- 12741572 TI - Technical note: Use of RFLP to characterize Lactococcus lactis strains producing exopolysaccharides. AB - Restriction fragment length polymorphism (RFLP) is used to differentiate microorganisms by analysis of their DNA restriction patterns. A modified RFLP procedure is proposed for the rapid characterization of Lactococcus lactis strains producing exopolysaccharides (EPS). The availability of such effective cataloging system is likely to benefit research aimed at identifying lactococcal strains that produce novel EPS. PMID- 12741573 TI - Genetic parameters for milk somatic cell scores and relationships with production traits in French Lacaune dairy sheep. AB - Records from 94,445 and 45,499 French Lacaune dairy ewes in first and second lactations, respectively, were used to estimate genetic parameters for somatic cell scores. Somatic cell count data came from an extensive recording scheme and sample testing that began in 1999 using the flocks enrolled in the official milk recording system. Somatic cell count data were from 2 to 4 test days per lactation. Lactation average and single test-day somatic cell scores were considered in multitrait sire models. The heritability estimate of lactation somatic cell score was close to 0.13 and similar for first and second parity. Heritabilities of somatic cell scores increased from first to fourth test day (from 0.07 to 0.11 in first lactation and from 0.05 to 0.13 in second lactation). Genetic correlations between somatic cell scores were high, usually more than 0.91, but lower between first test day and later test days in first lactation (0.64 to 0.88). The genetic correlations between lactation somatic cell score and milk yield, between lactation somatic cell score and fat content, and between lactation somatic cell score and protein content were 0.18, 0.04, and 0.03 in first lactation, respectively. The genetic antagonism between test day somatic cell score and milk yield measured in first lactation increased from beginning to the end of the lactation (0.05 to 0.23). This antagonism was slightly lower for somatic cell score in second lactation (from 0.09 to 0.14, and 0.08 for lactation mean). Environmental correlations in first lactation between lactation somatic cell score and milk yield, between lactation somatic cell score and fat content, and between lactation somatic cell score and protein content were -0.18, 0.13, and 0.30, respectively. PMID- 12741574 TI - Investigation of factors affecting voluntary and involuntary culling in expanding dairy herds in Wisconsin using survival analysis. AB - Trends in the relative risk of voluntary culling of low-producing cows and involuntary culling of high-producing cows were examined in 186 Wisconsin dairy herds that expanded significantly between 1994 and 1998. A Weibull model for survival analysis was applied to data of 72,456 Holstein cows with first calving from 1981 to 2000; this model included a time-independent effect of age at first calving and time-dependent effects of year-season, age-parity, and within herd year quintile for combined fat + protein yield (by time period). The relative risk of (involuntary) culling of high-producing cows (versus average cows) increased from 0.5 in 1981 to 1989 to 0.68 in 1996 to 2000. Meanwhile, the relative risk of (voluntary) culling of low-producing cows decreased from 4.20 to 2.55 over the same time period. Variables related to facilities, labor, and management were obtained via survey, and the relative risk of culling for high and low-producing cows after expansion (1996 to 2000) was calculated for different levels of each variable. Herds with fewer cows per employee and a greater percentage of labor supplied by family members tended to have lower risk of involuntary culling of profitable cows. Likewise, high-producing cows in herds with fans, sprinklers, self-locking manger stalls, palpation rails, and maternity pens had a significantly lower risk of culling than cows in herds without such facilities. Herds that used 100% artificial insemination (AI) had lower risk of involuntary culling than non-AI herds or herds with a cleanup bull, but 3x milking and use of a custom heifer grower led to unfavorable trends in involuntary culling. In summary, this study documented the unfavorable trends in voluntary and involuntary culling in expanding herds and quantified the gains producers can expect in cow survival by investing in improvements in facilities, labor, and management. PMID- 12741575 TI - Modifying the lactation curve to improve lactation milk and persistency. AB - Daily, stage and lactation estimated breeding values (EBV) and the shape of the lactation curve for each cow are controlled by a unique set of random (genetic) regression coefficients under a test day model, thus providing a basis for genetic improvement of these characteristics. Three selection procedures were developed for simultaneous improvement of total lactation milk and persistency: 1) index selection based on daily EBV, 2) index selection based on stage EBV, and 3) index selection based on random regression (RR) coefficients. A numerical example was given to demonstrate the computation of indexes based on stage EBV and based on RR coefficients. A conversion equation was derived to convert between genetic changes in EBV and RR coefficients. Index selection based on daily EBV would require the finding of 305 weighting factors for a lactation period of 305 d, making it impractical to determine the weighting factors on a daily basis. Alternatively, a lactation period was partitioned into a few stages to facilitate the construction of index selection based on stage EBV and index selection based on RR coefficients. These selection procedures make use of the annual genetic gains routinely computed in national genetic evaluations to restrict the genetic gains between different lactation stages to achieve the desired curve. When there is no prior knowledge of annual genetic gains, the proportional restriction of genetic gains between stages may be used. In summary, this study provides a simple means of modifying the lactation curve by manipulating genetic changes in different lactation stages at a pre-specified rate. PMID- 12741576 TI - Feeding behavior, milking behavior, and milk yields of cows milked in a parlor versus an automatic milking system. AB - This study compared feeding and milking behavior and milk yields for cows housed in the same barn, fed the same ration, but milked with a conventional milking parlor (parlor) or automatic milking system (robot). Behavioral data were videotaped hourly 1 d/mo for 9 mo. Feeding behavior patterns differed and were more variable for parlor cows than for robot cows. Both groups had low feeding rates at night and early morning. Feeding activity increased after milking and feed delivery for parlor cows. Milking and feeding activity in the robot system increased after human intervention at 7 a.m.; feed bunk activity peaked 3 h later and remained relatively constant from 10 a.m. to 8 p.m. Percentages of cows at the feed bunk were significantly greater for robot cows than parlor cows only at 10 a.m. and 9 p.m. Batch milking of parlor cows with free access to feed, vs. sequential milking of robot cows, with restricted movement to feed by a one-way gate system, resulted in higher peak percentages of cows at the bunk for parlor cows. Lower, more consistent percentages of cows eating at one time suggests that less bunk space may be needed for cows in robotic milking systems. Higher percentages of cows were observed in the robot from 8 a.m. to 1 p.m. and again from 3 to 7 p.m. Percentages of cows in the robot holding area were greatest from 8 to 11 a.m. and 3 to 6 p.m. and were lowest from midnight to 6 a.m. Milk production over 39 d in summer for subsets of cows was slightly but significantly higher (26.4 vs. 25.8 +/- 0.2 kg/d) for cows in the robot group. Milking frequency, days in milk, parity, and maximum air temperature for 3 d (-2 d to day of observation) affected milk yield comparisons. Results have implications for design of feeding and handling facilities used with automated milking systems. PMID- 12741578 TI - Timeliness and effectiveness of progeny testing through artificial insemination. AB - Progeny-test (PT) programs of US artificial-insemination (AI) organizations were examined to determine timeliness of sampling, PT daughter distribution, rate of return of PT bulls to widespread service, and genetic merit of PT bulls compared with AI-proven and natural-service (NS) bulls. Bull age at semen release and at birth and calving of PT daughters was documented by breed (Ayrshire, Brown Swiss, Guernsey, Holstein, Jersey, and Milking Shorthorn) for bulls that entered AI service since 1960. Mean Holstein bull age at semen release (16 mo) changed little over time, but standard deviations (SD) decreased from 4.0 mo during the 1960s to 2.4 mo during the 1990s. Most Holstein bulls (80%) had semen released by 18 mo. Mean age of Holstein bulls at birth and calving of PT daughters during the 1990s was 29 and 56 mo, respectively (a decline of 4 mo from the 1960s); SD decreased from 6 to 3 mo. Bulls of other breeds usually were older at birth and calving of PT daughters, and SD were larger. Mean Holstein bull age when 80% of PT daughters had been born declined from 36 mo during the 1960s to 31 mo during the early 1990s; for other breeds, bulls showed the same trend but at older ages. Mean Holstein bull age when 80% of PT daughters had calved declined from 65 mo during the 1960s to 59 mo during the 1990s; for other breeds, bulls were older. Percentage of herds with PT daughters has increased over time. For Holsteins, herds with five or more usable first-parity records that had PT daughters with usable records increased from 15% during 1965 to 61% during 1998; percentage of herds with from 1 to 19% PT records increased from 11 to 38%, and percentage of herds with >50% PT daughters increased from 1 to 5%. Percentage of Holstein PT bulls returned to AI service declined to about 12% for bulls with PT entry around 1990; for other breeds, 12 to 23% of most recent PT bulls were returned to service. Percentage of milking daughters that had records usable for genetic evaluation that were sired by PT bulls increased steadily from 10 to 18%, whereas percentage of daughters with usable records that were sired by NS bulls declined from 14 to 7%. Milk yield of daughters of AI-proven bulls was 107 to 200 kg greater than for daughters of PT bulls and 366 to 444 kg greater than for daughters of NS bulls for all years. More extensive and rapid sampling and increased selection intensity of PT programs have led to more rapid genetic progress. More extensive use of AI could increase US producer income by millions of dollars annually. PMID- 12741577 TI - Effect of on-farm commercial batch pasteurization of colostrum on colostrum and serum immunoglobulin concentrations in dairy calves. AB - The objectives were to describe the effect of on-farm commercial batch pasteurization on immunoglobulin (IgG) concentrations and the fluid and feeding characteristics of colostrum and to compare serum IgG concentrations in calves fed fresh versus pasteurized colostrum. Newborn calves (123) were systematically allocated to dietary treatments of either fresh or pasteurized colostrum at both the first and second colostrum feedings. The IgG concentrations were measured for batches of colostrum fed fresh and in pre and postpasteurized samples for batches of colostrum fed after being pasteurized and in calf serum. Pasteurization reduced colostrum IgG concentration, with the percentage reduction averaging 58.5 and 23.6% for 95-L and 57-L batches, respectively. Pasteurizing high quality colostrum in 57-L (vs. 95-L) batches resulted in higher IgG concentrations in the end product. Pasteurization of 57-L batches produced colostrum of normal or only mildly thickened consistency that could be fed to calves. Serum IgG concentrations were higher for calves fed fresh colostrum and for calves with a shorter time interval (< or = 6 h) between first and second colostrum feedings. After controlling for the time interval between feedings, serum IgG concentrations were significantly higher for 40 calves fed unpasteurized (19.1 mg/ml) vs. 55 calves fed pasteurized colostrum (9.7 mg/ml) for calves fed 2 L at first feeding. By contrast, there was no difference in serum IgG concentrations between 8 calves fed unpasteurized (16.1 mg/ml) and 20 calves fed pasteurized colostrum (13.5 mg/ml) after calves were fed 4 L at the first feeding. While the latter results suggest that pasteurizing colostrum may work for producers with excellent colostrum management, these results are preliminary and should be interpreted with caution, given the fewer number of calves and batches of colostrum involved with this second comparison. PMID- 12741579 TI - HIPAA privacy rule and public health. Guidance from CDC and the U.S. Department of Health and Human Services. AB - New national health information privacy standards have been issued by the U.S. Department of Health and Human Services (DHHS), pursuant to the Health Insurance Portability and Accountability Act of 1996 (HIPAA). The new regulations provide protection for the privacy of certain individually identifiable health data, referred to as protected health information (PHI). Balancing the protection of individual health information with the need to protect public health, the Privacy Rule expressly permits disclosures without individual authorization to public health authorities authorized by law to collect or receive the information for the purpose of preventing or controlling disease, injury or disability including but not limited to public health surveillance, investigation, and intervention. Public health practice often requires the acquisition, use, and exchange of PHI to perform public health activities (e.g., public health surveillance, program evaluation, terrorism preparedness, outbreak investigations, direct health services, and public health research). Such information enables public health authorities to implement mandated activities (e.g., identifying, monitoring, and responding to death, disease, and disability among populations) and accomplish public health objectives. Public health authorities have a long history of respecting the confidentiality of PHI, and the majority of states as well as the federal government have laws that govern the use of, and serve to protect, identifiable information collected by public health authorities. The purpose of this report is to help public health agencies and others understand and interpret their responsibilities under the Privacy Rule. Elsewhere, comprehensive DHHS guidance is located at the HIPAA website of the Office for Civil Rights (http://www. hhs.gov/ocr/hipaa/). PMID- 12741580 TI - Effect of external sequential compression devices on femoral venous blood flow. AB - Sequential compression devices are used to reduce venous stasis and deep venous thrombosis after joint replacement. Thigh-length, calf-length, and foot compression devices were compared in using ultrasonography after unilateral knee arthroplasty. Simulated muscle activity via active ankle motion was also evaluated. Blood flow volume and velocity were recorded above and below the saphenous vein bifurcation, the division of the superficial and deep systems, allowing evaluation of each. Volume and velocity increased in the superficial and deep systems with all devices. A control group was evaluated to determine differences related to age and surgery. The devices performed similarly in the volunteers. However, active motion performed better than any device. Thus, unlike young, healthy patients, muscle activity alone in the operative population was unreliable in increasing blood flow. Thigh-length, calf-length, and foot compression devices are are effective at increasing femoral blood flow volume and velocity in the deep and superficial venous systems after total knee arthroplasty. PMID- 12741582 TI - Radiographic evaluation of periprosthetic metallosis after total knee arthroplasty. AB - This retrospective study examined the clinical significance of a radiographic sign associated with periprosthetic metallosis after total knee arthroplasty. Of 71 knees undergoing revision arthroplasty over an 8-year period, 11 had gross evidence of metal debris identified intraoperatively. Histologic preparations confirmed the presence of particulate metal debris in all cases. Radiographs in 7 of 11 cases were positive for metallosis. The radiographic sign identified on lateral radiographs was divided into 3 types based on the size of the mass. The magnitude of soft tissue pathology and the extent of osteolysis correlated with the size of the mass on preoperative radiographs. Replacement of all components was necessary in 71% of cases with radiographs positive for metallosis and 47% of cases with negative radiographs. Only 1 of the 11 knees with metallosis had a late postoperative infection, for which 2-stage revision arthroplasty was required. All cases with positive radiographs had gross and histologic confirmation of metal and polyethylene debris. These data suggest that careful assessment of radiographs can facilitate preoperative planning. PMID- 12741581 TI - Biomechanical comparison of reconstruction techniques for disruption of the acromioclavicular and coracoclavicular ligaments. AB - Injuries to the acromioclavicular joint are common. For selected injuries, operative reconstruction is recommended. The purpose of the current study was to compare three reconstruction procedures: (1) nine strands of woven polydioxanonsulphate (PDS II) suture passed through the clavicle and around the coracoid; (2) procedure No. 1 with 50% of the coracoacromioclavicular ligament placed through 2 clavicular drill holes; (3) No. 5 Merselene tape passed through 2 drill holes in the clavicle and acromion, with 50% of the coracoacromial ligament transferred to the clavicle. Fourteen fresh frozen human shoulders were tested using a 6 degree-of-freedom testing device. The intact shoulder showed significantly less displacement than any of the reconstructions. Merselene tape plus ligament showed the largest displacement, and PDS II braid plus ligament showed the least displacement. None of the procedures reconstituted acromioclavicular joint stiffness to intact state levels, though improved acromioclavicular joint stiffness was noted with a PDS braid plus ligament. PMID- 12741583 TI - Revision anterior cruciate ligament reconstruction surgery. AB - In 1995, it was reported that 60,000 to 75,000 anterior cruciate ligament (ACL) reconstructions were being performed annually in the United States. Successful long-term results are achieved in 75% to 95% of these patients, but 8% have unsatisfactory results due to recurrent instability and graft failure. With the increasing popularity of this procedure, ACL revision surgery has also become increasingly common. While the techniques described for ACL revision have been varied, the overall results in the literature do not compare favorably with primary ACL reconstruction. The proper execution of revision ACL reconstruction requires precise preoperative planning to assess the cause of initial failure and avoid repeating the same mistakes with revision reconstruction. Graft choice, hardware removal, revision notchplasty, tunnel placement, and method of fixation are key points for a successful result. The causes of ACL failure, the technical aspects of revision ACL surgery, and the reported results of revision ACL surgery are reviewed. PMID- 12741584 TI - Fresh osteochondral grafting in the treatment of osteochondritis dissecans of the talus. AB - We present a review of the literature on classification and treatment of osteochondral defects of the talus. We report the case of an isolated Berndt and Harty grade II lesion treated with a fresh osteochondral allograft. We believe that fresh allograft osteochondral grafting of the talus is an excellent technique for symptomatic Berndt and Harty grade II or higher lesion of the talus without significant tibiotalar arthritis. In selected patients, this procedure can provide excellent functional results. PMID- 12741585 TI - Proximal femoral allografting in revision total hip arthroplasty: stabilization of the host-graft junction with tension band fixation. AB - Large deficiencies of the metaphysis and proximal diaphysis present a significant challenge to the orthopaedic surgeon undertaking revision of failed total hip arthroplasty. Proximal femoral allograft-prosthetic reconstruction may enable revision in such situations. Secure fixation is essential to successful integration and function of these allograft-prosthetic constructs. We report our experience in 7 revision total hip arthroplasties in 5 patients. Proximal femoral allograft-prosthetic reconstruction was undertaken on all 7 hips using a technique of tension band fixation for securely fixing allograft to host bone at their interface to further minimize motion at the junction site and maximize union. Radiographic evidence of union at the interface was noted in 6 of 7 hips. Mean Harris hip score increased from 18 preoperatively to 83 postoperatively, with a mean follow-up of 78 months (range, 56-116 months). PMID- 12741586 TI - Cost impact of botulinum toxin use in Medicaid-enrolled children with cerebral palsy. AB - The use of botulinum toxin type A (BTX) in the management of spasticity in childhood cerebral palsy (CP) is increasing. This study examined annual health care service utilization and costs associated with BTX therapy for spastic CP in Medicaid-enrolled children receiving complete health care coverage (1997 to 1999). We used pair matching as well as recent statistical technique improvements (bootstrap method) to work with limited samples. The introduction of BTX was associated with an increase of approximately $62 per month in prescription costs for the patient. However, these costs were made up by reductions in hospitalization. When each year was examined individually, reimbursements for BTX users were not different from those for pair-matched non-BTX users. These data suggest that BTX therapy does not significantly add to the costs of treating Medicaid-enrolled children with CP. PMID- 12741587 TI - Determination of bone mineral content in cadaveric test specimens. AB - This study was designed to determine the best method for presenting the bone mineral content of cadaveric test specimens. A total of 59 bone samples were taken from the humeri, radii, and ulnae of 14 female cadavers. For each sample, the bone mineral content was determined with a dual-energy x-ray absorptiometry scanner, and the ash-weight ratio was calculated manually. A linear regression analysis was performed to compare the ash-weight ratio to the 3 methods for reporting bone mineral content as measured by a dual-energy x-ray absorptiometry scanner: by bone mineral content divided by sample length (in g/cm), by projected area (in g/cm2), or by sample volume (in g/cm3). The analysis revealed that the ash-weight ratio correlates best with the volumetric representation. Based on these data, a volumetric representation is suggested as the best representation of bone mineralization, due to its correlation with ash-weight ratio. PMID- 12741588 TI - Pigmented villonodular synovitis of the knee: average five-year follow-up of arthroscopic treatment. AB - We have reviewed the results of arthroscopic treatment of pigmented villonodular synovitis (PVNS) with reference to both recurrence and to function. Between 1985 and 1995, a single surgeon treated eight patients. At an average 5-year follow up, all patients were interviewed and had assessment of Hospital for Special Surgery (HSS) knee score for both the affected and unaffected knees. Also recorded were age, sex, and whether disease was recurrent, localized, or diffuse. Disease recurred in 4 patients, all with diffuse PVNS, and 3 of them required a further arthroscopic synovectomy at a mean of 16 months after the index procedure. All patients had good or excellent functional results. There was no significant difference between HSS knee scores for affected and unaffected knees. Arthroscopic synovectomy is a successful treatment in patients with localized PVNS of the knee and results in a knee that is functionally not different from its unaffected partner. PMID- 12741589 TI - Cerebral palsy: past, present, and future. PMID- 12741590 TI - Thermal capsulorrhaphy in the treatment of multidirectional instability of the shoulder. AB - The treatment of symptomatic multidirectional instability (MDI) of the shoulder has proven a challenge to orthopaedic surgeons. Patients who refuse activity modification often fail nonsurgical management, and traditional surgical approaches may trade the dysfunction of instability for that of decreased motion and possible future arthrosis. Recently, several methods of arthroscopic thermal capsulorrhaphy have gained popularity. Studies have shown that capsular shrinkage occurs due to the denaturation of type I collagen with maintenance of heat-stable intermolecular cross-links. Thermally treated tissues undergo a predictable healing response. The primary advantages of this procedure are the decreased surgical morbidity and ease of procedure compared with open capsular shift. Additionally, it may provide improved stability and decreased pain while maintaining range of motion. When radiofrequency treatment provides inadequate visualized capsular contracture, arthroscopic rotator interval closure can improve stability without resorting to an open procedure. While initial results in certain clinical series are promising, the long-term efficacy remains to be seen. PMID- 12741591 TI - History of orthopaedic education. PMID- 12741592 TI - Transient erectile dysfunction associated with intramuscular injection of botulinum toxin type A. AB - Autonomic nervous system dysfunction occurs rarely after botulinum toxin type A (BTX-A) intramuscular injections. We report a case of a 23-year-old man with spastic diplegia who had transient erectile dysfunction after intramuscular injection of BTX-A (total dosage, 300 IU, body weight 95 kg) in both hamstring muscles. Some investigators believe that the local spread of the toxin is responsible for autonomic dysfunction, while others believe that the transportation of the toxin to the spinal cord via retrograde flow or via the blood flow after entering the circulation are possible mechanisms of neurologic side effects. On the basis of our case, a retrograde axoplasmic flow to the spinal cord could probably occur because the spinal cord level of hamstring muscles is close to spinal cord levels responsible for erection control. PMID- 12741594 TI - 2002 Distinguished Southern Orthopaedist Award. PMID- 12741593 TI - Chronic osteomyelitis associated with cat-scratch disease. AB - Cat-scratch disease (CSD) is usually a self-limited illness, though atypical presentations of infection with Bartonella henselae can occur, including osteomyelitis, oculoglandular syndrome, and granulomatous hepatitis. We describe a 6-year-old boy who had atypical CSD osteomyelitis of the left proximal femoral metaphysis due to a cat scratch. This is the second paper to report serial serology of B henselae, and the second paper to identify plasma cells on histologic examination, compatible with chronic osteomyelitis. The diagnosis was made by clinical, serologic, and histologic examination. Sixteen cases of atypical CSD osteomyelitis have been reported in the English literature and are reviewed in this paper. PMID- 12741595 TI - Selective separation of active inhibitors of epidermal growth factor receptor from Caragana jubata by molecularly imprinted solid-phase extraction. AB - A feasibility research was performed to study the possibilities of using a molecularly imprinted polymer as sorbent material in solid-phase extraction for the separation of active inhibitors of epidermal growth factor receptor (EGRF) from Caragana Jubata, a Chinese traditional Tibetan medicine. A molecularly imprinted polymer using quercetin, an active anti-EGFR inhibitor (IC50 = 15 microM), as the template and acrylamide as the functional monomer was prepared. The polymer was evaluated as a selective sorbent in molecularly imprinted solid phase extraction. The EtOAc extract of Caragana Jubata was loaded on the polymer, and two novel active anti-EGFR inhibitors were found to be selectively retained after washing the polymer with appropriate solvent to disrupt the non-specific interactions occurring between the sample and the polymer matrix, which were identified as (E)-piceatannol (IC50 =4.9 microM) and butein (IC50 = 10 microM). The present work affords us a new potential method for selective separation of bioactive components from herb by using molecularly imprinted polymer as a solid phase extraction adsorbent. PMID- 12741596 TI - Microscale solid-phase extraction system for explosives. AB - A simple, semi-automated, microcolumn solid-phase extraction (SPE) system is optimized for the extraction, preconcentration and HPLC analysis of seven different explosives and explosive derivatives contaminating seawater, river water and well water samples. The microcolumns were constructed from 1/16 in. O.D. PTFE tubing (1 in.=2.54 cm) packed with 0.5-1.5 mg of SPE material. LiChrolut EN or Porapak R. The extraction system consisted of two syringe pumps and several solenoid valves. Optimal detection limits were realized when the sample water flow-rate was maximally increased within the limits of the pump, 5 10 ml/min (despite exceeding the breakthrough threshold of the SPE microcolumn), and when the eluate volume collected from the column was minimized, <5 microl (despite very low recovery percentages). PMID- 12741597 TI - Computational fluid dynamics simulations yielding guidelines for the ideal internal structure of monolithic liquid chromatography columns. AB - A theoretical calculation of the separation performance of a (hypothetical) micro structured monolithic LC column is presented, confirming that the polydispersity effect in parallel bundle columns can theoretically be eliminated to a very large extent by radially redistributing the mobile phase fluid at regular intervals. It is demonstrated that the flow can be redistributed in such a way that the advantage coming from the suppression of the polydispersity effect largely exceeds the losses caused by the additional pressure-drop and band broadening. The presently considered micro-structured column would allow to perform N > 100,000 plate separations in a few hundred of seconds, i.e., about an order of magnitude faster than the best possible packed bed and monolithic HPLC columns, while offering the same mass loadability. This clearly demonstrates that the currently available LC columns are still far away from the absolute resolution limit of the ideal, fully optimised LC column. PMID- 12741598 TI - Safety concerns in ultrahigh pressure capillary liquid chromatography using air driven pumps. AB - Ultrahigh pressure liquid chromatography (UHPLC) is an emerging technique which utilizes pressures higher than 10,000 p.s.i. to overcome the flow resistance imposed when using very small particles as packing materials in fused-silica capillary columns (1 p.s.i.=6894.76 Pa). This technique has demonstrated exceptionally high separation speeds and chromatographic efficiencies. However, safety is a concern when extremely high pressures are used. In this study, the safety aspects of capillary column rupture during operation were identified and carefully evaluated. First, liquid jets may be formed as a result of blow-out of the on-column frits or from rupture of the capillary at or near the column inlet. Second, incorrect installation of the capillary at the injector, failure of the ferrule used in the capillary connection, or rupture of the capillary can produce high speed projectiles of silica particles or column fragments. Experiments were carried out in the laboratory to produce liquid (water) jets and capillary projectiles using a UHPLC system, and the power density, an important parameter describing water jets in industrial practice, was calculated. Experimental results were in accordance with theoretical calculations. Both indicated that water jets and capillary projectiles under ultrahigh pressures might lead to skin penetration under limited conditions. The use of a plexiglass shroud to cover an initial length of the installed capillary column can eliminate any safety-related concerns about liquid jets or capillary projectiles. PMID- 12741599 TI - Chemical compositional separation of styrene-methyl methacrylate copolymers using high-performance liquid chromatography with liquefied carbon dioxide as eluent. AB - Chromatographic separation of styrene-methyl methacrylate (MMA) copolymers depending on the chemical composition was studied using liquefied carbon dioxide as an adsorption promoting solvent, and tetrahydrofuran, chloroform containing ethanol as a desorption promoting solvent in the mobile phase and the column packed non-bonded silica gel by a solvent gradient method. With the increase of MMA content, the elution was retarded indicating that the typical normal-phase type of adsorption occurred. The effects of type of desorption solvents, molecular mass of sample, and column temperature on the elution were investigated. PMID- 12741600 TI - Comparison of gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry for the determination of fatty and resin acids in paper mill process waters. AB - A comparative study of the performance of liquid chromatography (LC)-atmospheric pressure chemical ionisation (APCI)-mass spectrometry (MS) and gas chromatography (GC)-mass spectrometry techniques for the determination of resin and fatty acids from paper mill process waters was carried out. These compounds are responsible for the high toxicity of paper mill effluents and little research has been carried out regarding their analysis using mass spectrometric techniques. To prove the usability of GC and LC-MS, 16 treated and untreated water samples of recycle, kraft and pulp paper mills were analysed and good agreement was observed as regards to compounds detected and corresponding concentrations. This paper also reports the limits of detection, recoveries, reproducibility, linearity and precision using the two methods. GC-MS presented better selectivity and lower detection limits (below 0.2 microg/l), but derivatization of the extracts and the short life of derivatives (12-24 h) made the technique tedious and prone to high variations. Although LC-APCI-MS presented coelution of the non-aromatic resin acids, it also showed good sensitivity (limits of detection <3 microg/l) and permitted the detection of resin and fatty acids at microg/l level. In addition, since samples could be directly injected to the chromatographic system, LC-APCI MS was proven as a powerful technique for quick and unequivocal quality control during papermaking. PMID- 12741601 TI - Extending the range of compounds amenable for gas chromatography-mass spectrometric analysis. AB - Gas chromatography-mass spectrometry (GC-MS) suffers from a major limitation in that an expanding number of thermally labile or low volatility compounds of interest are not amenable for analysis. We found that the elution temperatures of compounds from GC can be significantly lowered by reducing the column length, increasing the carrier gas flow rate, reducing the capillary column film thickness and lowering the temperature programming rate. Pyrene is eluted at 287 degrees C in standard GC-MS with a 30 m x 0.25 mm I.D. column with 1-microm DB5ms film and 1-ml/min He column flow rate. In contrast, pyrene is eluted at 79 degrees C in our "Supersonic GC-MS" system using a 1 m x 0.25 mm I.D. column with 0.1-microm DB5ms film and 100-ml/min He column flow rate. A simple model has been invoked to explain the significantly (up to 208 degrees C) lower elution temperatures observed. According to this model, every halving of the temperature programming rate, or number of separation plates (either through increased flow rate or due to reduced column length), results in approximately 20 degrees C lower elution temperature. These considerably lower elution temperatures enable the analysis of an extended range of thermally labile and low volatility compounds, that otherwise could not be analyzed by standard GC-MS. We demonstrate the analysis of large polycyclic aromatic hydrocarbons (PAHs) such as decacyclene with ten fused rings, well above the current GC limit of PAHs with six rings. Even a metalloporphirin such as magnesiumoctaethylporphin was easily analyzed with elution temperatures below 300 degrees C. Furthermore, a range of thermally labile compounds were analyzed including carbamates such as methomyl, aldicarb, aldicarbsulfone and oxamyl, explosives such as pentaerythritol tetranitrate, Tetryl and HMX, and drugs such as reserpine (608 a.m.u.). Supersonic GC-MS was used, based on the coupling of a supersonic molecular beam (SMB) inlet and ion sources with a bench-top Agilent 6890 GC plus 5972 MSD. The Supersonic GC-MS provides enhanced molecular ion without any ion source related peak tailing. Thus, the lower GC separation power involved in the analysis of thermally labile and low volatility compounds is compensated by increased separation power of the MS gained from the enhanced molecular ion. Several implications of these findings are discussed, including our conclusion that slower chromatography leads to better analysis of thermally labile compounds. PMID- 12741602 TI - Large-volume programmed-temperature vaporiser injection for fast gas chromatography with electron capture and mass spectrometric detection of polybrominated diphenyl ethers. AB - A large volume injection fast-GC-MS method has been developed, optimized and evaluated for the determination of polybrominated diphenyl ethers, including the decabrominated diphenyl ether (BDE-209). The programmed-temperature vaporiser injection parameters, temperature programming of the GC oven, and the physical dimensions of the narrow bore GC column were investigated to find the optimal operating conditions for the analysis. Depending on parameter settings the yield of the PBDEs and particularly BDE-209, varies significantly. Volumes up to 125 microl were successfully injected and a fast GC separation was performed, with retention times as short as 6.4 min for the last eluting compound, BDE-209. In a pilot study an air sample, collected at an electronics dismantling facility, was analyzed. Low-resolution mass spectrometry in electron capture negative ion mode was used for detection. Nine BDE congeners, including BDE-209, were identified and quantified. PMID- 12741603 TI - Optimized conditions for hydrocarbon group type analysis of base oils by thin layer chromatography-flame ionisation detection. AB - The results of research on the optimization of the thin-layer chromatography flame ionisation detection for the determination of group composition of natural base oils, including separation of the aromatics into subgroups, are presented. Neutral base oils obtained in several steps of refining from vacuum distillation petroleum fractions are the most difficult to analyze by hydrocarbon group type analysis (HGTA) because of the high content of aliphatic fragments in their molecules. Factors affecting the accuracy and precision of the results were identified. The paper presents the analytical procedure, including two different calibration methods, as well as the results of studies on the reproducibility of HGTA of typical base oils of different viscosity classes under the optimized conditions. The same conditions were found suitable for HGTA of other high boiling petroleum fractions by TLC with flame ionisation detection. The paper also introduces a new procedure for reproducible determination of polar fractions in base oils utilizing solid-phase extraction columns, and presents a corrected procedure for the determination of saturated compounds and aromatics (mono-, bi- and polycyclic) in base oils by column liquid chromatography. PMID- 12741604 TI - Rotation planar extraction and rotation planar chromatography of oak (Quercus robur L.) bark. AB - The versatile novel instrument for rotation planar extraction and rotation planar chromatography was exploited for the investigation of oak bark (Quercus robur L.). The same instrument enabled extraction of the bark, analytical proof of (+) catechin directly in the crude extract and also its fractionation. Additionally, epimeric flavan-3-ols, (+)-catechin and (-)-epicatechin were separated by analytical ultra-micro rotation planar chromatography on cellulose plates with pure water as developing solvent. A comparison of the extraction of oak bark with 80% aqueous methanol by rotation planar extraction and medium pressure solid liquid extraction was carried out and both techniques were shown to be suitable for the efficient extraction of oak bark. The raw extracts and fractions on thin layer chromatography showed many compounds that possessed antioxidant activity after spraying with 1,1-diphenyl-2-picrylhydrazyl. Rotation planar fractionation of 840 mg of crude oak bark extract on silica gel gave 6.7 mg of pure (+) catechin in one run. PMID- 12741605 TI - Separation and quantitation of components in FD&C Red No. 3 using capillary electrophoresis. AB - The use of capillary electrophoresis as a technique to separate and quantitate components of FD&C Red No. 3 (erythrosine, color index No. 45430) is described. The fluorescein isomers, 2',4',5'-triiodofluorescein (2,4,5-I3F) and 2',4',7' triiodofluorescein (2,4,7-I3F), the most abundant by-products formed during the preparation of the dye, were selected for quantitation studies. The separation of other lower halogenated impurities was also demonstrated. Electrophoretic mobility of the compounds was achieved in a 50 mM borate, 25 mM sodium dodecyl sulfate buffer at pH 9.3. The limits of quantitation were found to be 0.15% (w/w) (2,4,5-I3F) and 0.14% (w/w) (2,4,7-I3F) (relative to the mass of FD&C Red No. 3). The method is linear from 0.08 to 20.0% (w/w) for 2,4,5-I3F and between 0.06 and 17.0% (w/w) for 2,4,7-13F. In addition, relative standard deviations of 2.03 and 5.11% were determined from precision studies in the repeat analysis of FD&C Red No. 3 for 2,4,5-I3F and 2,4,7-I3F, respectively. Overall, the CE method produced data in excellent agreement with the reference HPLC method, used considerably less solvent and sample, generated less waste and was found to be considerably more cost efficient. PMID- 12741606 TI - Computer-assisted method development and optimization in high-performance liquid chromatography. AB - This paper reports the use of DryLab, a computer simulation software package, to assist in the development and optimization of a reversed-phase high-performance liquid chromatographic (HPLC) method for the separation of a model drug candidate and its degradation products. Prior to the optimization process, columns with various bonded phases are evaluated for their chromatographic performance using the sample of interest. Simultaneous optimization of two separation variables and the use of resolution maps to predict the optimal conditions are illustrated. Options to optimize column conditions (column length and flow-rate) to further reduce run time are briefly discussed. The accuracy of DryLab-predicted retention times and resolution is compared with experimental values. The DryLab software used in this study provided satisfactory predictions for the selected model, with average errors of less than 3.5 and 11.8% for retention time and resolution, respectively. PMID- 12741607 TI - New developments in insulin delivery. AB - A vigorous research effort has been undertaken worldwide to replace injectable insulin by a more comfortable and painless delivery method. Several routes have been explored for their suitability with respect to insulin degradation in the human body. Considerable progress has been made in achieving the common goal for a convenient and equally effective insulin delivery. This article reviews the different routes available for insulin administration and the many successful developments that have been made in recent years for improving that particular route for a much better insulin delivery. PMID- 12741608 TI - Improvement of the low-temperature stability of an aqueous colloidal ethylcellulose dispersion, Aquacoat ECD, and preparation/characterization of a redispersible Aquacoat ECD powder. AB - Aquacoat ECD, an aqueous ethylcellulose dispersion, is susceptible to low temperature storage, resulting in irreversible coagulation as indicated by a strong increase in viscosity and particle size. This destabilization of the ethylcellulose dispersion is caused by the anionic surfactant sodium dodecyl sulfate, which precipitates at low temperatures because of its low Krafft point. This problem could be solved by using the more hydrophilic, ethoxylated, sodium dodecyl ether sulfate, which was an effective stabilizer at low temperatures. A redispersible ethylcellulose powder was prepared by freeze- or spray-drying of the original polymer dispersion (Aquacoat ECD). The pH of the dispersion medium had a strong influence on the redispersibility of the dried ethylcellulose particles because of the dissociation behavior. At a pH > 7, polymer dispersions in the colloidal size range were obtained. At lower pH values, the dried ethylcellulose agglomerates could not be redispersed. Drug release studies from pellets coated with the redispersed and the original dispersion showed a comparable release pattern when using alkalized aqueous dispersion systems. Higher plasticizer concentration and curing of the coated pellets improved the film formation of the redispersed polymer particles. PMID- 12741609 TI - Investigating the effects of excipients on the powder flow characteristics of theophylline anhydrous powder formulations. AB - Pharmaceutical excipients may have a great effect on properties affecting tablet production. To determine if formulations containing theophylline anhydrous would have properties allowing them to be easily tableted, functional parameters affecting powder flow were evaluated. The Carr Flowability Indices were used for this evaluation. Formulations to be studied include theophylline anhydrous as the active ingredient, hydrous lactose and dicalcium phosphate dihydrate as diluents, polyvinylpyrrolidone as a binder, and fumed silica as a flow promoter. The effect of each component on powder flow is discussed. PMID- 12741610 TI - Effect of polymer hydration on the kinetic release of drugs: a study of ibuprofen and ketoprofen in HPMC matrices. AB - Samples of drug/hydroxypropylmethylcellulose (HPMC) mixtures and matrices (drug/HPMC mixtures plus excipients) were allowed to equilibrate in closed chambers with defined relative humidities (RHs). Their water uptake and drug release were evaluated by differential scanning calorimetry/thermogravimetric analysis and dissolution studies, respectively. Analysis of the thermal behaviors of the drug/HPMC mixtures and of the polymer alone, as functions of RH, leads to the conclusion that most of the hydration water is retained by the polymer, and points to the occurrence of different types of hydration water, from the strongly polymer-bound water molecules at RH values up to 81%, to the almost "free water" for RH values close to 100%. In addition, application of the Korsmeyer model to the dissolution results leads to the conclusion that the rate determining dissolution processes are predominantly of the fickian type. PMID- 12741611 TI - Processing of nimesulide-PEG 400-PG-PVP solid dispersions: preparation, characterization, and in vitro dissolution. AB - The objective of this investigation was to study the influence of dissolution enhancers such as polyethylene glycol 400, propylene glycol, polyvinylpyrrolidone K30, sodium lauryl sulfate, and Tween 80 on in vitro dissolution of a model active pharmaceutical material--nimesulide. Preliminary studies were conducted using a physical blend of nimesulide, and the adjuvants and solid dispersions were prepared using solvent evaporation and cogrinding methods. Aqueous solution of adjuvants was first triturated with nimesulide, followed by mixing with lactose and microcrystalline cellulose, and finally water was evaporated under vacuum in a cogrinding method. A 33 factorial design was adopted in a cogrinding method using the concentration of polyethylene glycol 400, propylene glycol, and polyvinylpyrrolidone K30 as independent variables. Tween 80 and sodium lauryl sulfate were added in all the batches. Full and reduced models were evolved for different dependent variables. The reduced models were validated using two checkpoints. Angle of repose < 35 degrees, percentage of drug released in 30 min (Q30) > 40%, 45 min (Q45) > 50%, and 120 min (Q12) > 60% were used as constraints for the selection of an optimized batch. Contour plots are presented for the selected dependent variables. Polyvinylpyrrolidone was found to be more effective in increasing the drug dissolution, compared with polyethylene glycol 400 and propylene glycol. The granule flow was adversely affected when high levels of liquid adjuvants were used in formulations. Wettability study was conducted to measure wetting time for pure drug and the optimized batch. Improved drug dissolution was attributed to improved wetting and the solubilizing effect of adjuvants from the pseudosolid dispersions of nimesulide. Significant improvement in drug dissolution was observed (Q120 = 70%), compared with pure drug powder (Q120 = 15%). In conclusion, dissolution of nimesulide can be modulated using an appropriate blend of pharmaceutical adjuvants. PMID- 12741612 TI - Evaluation of the effects of khaya gum on the mechanical and release properties of paracetamol tablets. AB - A study of the comparative effects of khaya gum and two standard binding agents polyvinylpyrrolidone (PVP) and gelatin--on crushing strength and friability, and the disintegration and dissolution characteristics of paracetamol tablets was made. The crushing strength-friability ratio (CSFR), the disintegration times, D, and the dissolution times t50, t90, and t1 (derived from the equation of Noyes and Whitney), all increased with an increase in binder concentration; however, the dissolution rate constants, k1 and k2, decreased. The ranking for the values of CSFR for tablets containing the different binders was PVP > gelatin > khaya gum. The ranking for D and the dissolution times was gelatin > khaya gum > PVP, whereas the ranking for the dissolution rate constants was PVP > khaya gum > gelatin. There were significant linear correlations between CSFR, D, t50, t90, and t1 for the tablets. There were also significant correlations between k1 and D, t50, t90, and t1, and between k2 and t90. The results suggest that khaya gum could be useful as an alternative binding agent to produce tablets with particular mechanical strength and drug release profiles. PMID- 12741613 TI - Preparation and in vitro/in vivo evaluation of the buccal bioadhesive properties of slow-release tablets containing miconazole nitrate. AB - Slow-release buccal bioadhesive tablets of miconazole nitrate were prepared by using polymer mixtures of buccoadhesive materials such as hydroxypropylmethylcellulose, sodium carboxymethylcellulose, carbopol 934p, and sodium alginate. The physicochemical properties, swelling index, microenvironment pH, in vitro drug release, in vivo buccoadhesion time, and miconazole salivary concentrations of the prepared tablets were shown to be dependent on the type and composition of the buccoadhesive materials used. The dissolution of miconazole from all the prepared tablets into phosphate buffer (pH 6.8) was controlled and followed non-Fickian release mechanisms. All the prepared tablets gave reasonable buccoadhesion time (2.45-3.65 hr). Infrared spectroscopy and differential scan calorimetry studies revealed the absence of significant interactions between miconazole nitrate and the selected buccoadhesive materials. Duration of the antifungal activity as measured by the inhibition zone of Candida albicans by extracted human saliva was significantly longer (p < 0.05), compared with commercial miconazole oral gel (Daktaren oral gel). Based on the results obtained, the prepared slow-release buccoadhesive tablets of miconazole would markedly prolong the duration of the antifungal activity with more patient convenience. PMID- 12741614 TI - Physicochemical characterization and drug release studies of nilvadipine solid dispersions using water-insoluble polymer as a carrier. AB - Nilvadipine solid dispersions were prepared by the solvent method using water insoluble polymers, including low-substituted hydroxypropylcellulose, croscarmellose sodium, carmellose calcium, carmellose, and crospovidone. Differential scanning calorimetry and powder x-ray diffraction analysis showed that nilvadipine was present in an amorphous state in the solid dispersion obtained using crospovidone as a carrier. The degree of crystallinity of nilvadipine was dependent on the ratio of nilvadipine to crospovidone, and nilvadipine was present in an amorphous state when the ratio of nilvadipine to crospovidone was below one-half. Fourier transform infrared studies suggested the presence of hydrogen bonding between nilvadipine and crospovidone in the solid dispersion. Dissolution studies indicated that the maximum percentage of dissolution and dissolution rate constants were markedly increased in nilvadipine with crospovidone solid dispersion, compared with those of pure nilvadipine and physical mixtures. The dissolution rate of nilvadipine solid dispersion with crospovidone could be calculated by the Higuchi square root time equation. PMID- 12741615 TI - Solubilization of (+)-limonene by anionic/cationic mixed surfactant systems. AB - The combined effects of sodium n-octyl sulfate (SOS) and cetyltrimethylammonium bromide (CTAB) on the solubilization of (+)-limonene in aqueous solution were studied using a headspace gas chromatography technique. The findings showed the mixing of SOS and CTAB resulted in positive synergistic effects on the solubility of (+)-limonene. The positive synergistic effects are explained from the perspective of the phase behavior of this mixed surfactant system. PMID- 12741617 TI - Influence of Eudragit NE 30 D blended with Eudragit L 30 D-55 on the release of phenylpropanolamine hydrochloride from coated pellets. AB - The objective of this study was to investigate the influence of Eudragit NE 30 D blended with Eudragit L 30 D-55 on the release of phenylpropanolamine hydrochloride (PPA x HCl) from coated pellets. The miscibility of Eudragit NE 30 D/L 30 D-55 blends at different ratios was studied by using differential scanning calorimetry. The release of PPA x HCl from pellets coated with Eudragit NE 30 D alone and a Eudragit NE 30 D/L 30 D-55 blend, when stored at 40 degrees C and 60 degrees C, was determined by UV spectroscopy. Eudragit NE 30 D and Eudragit L 30 D-55 were miscible in ratios greater than 4:1. The curing time that was required to reach an equilibrium state decreased with the addition of Eudragit L 30 D-55. The presence of Eudragit L 30 D-55 also produced a film coating that was less tacky, and a dispersion of Eudragit NE 30 D containing Eudragit L 30 D-55 (5:1) was shown to prevent agglomeration of the pellets during coating and storage. PMID- 12741616 TI - Comparison of neural network and multiple linear regression as dissolution predictors. AB - The predictive performance of an artificial neural network (NN) was compared with the first-order multiple linear regression (MLR) using mean dissolution data of 28 diltiazem immediate release tablet formulations. The performance was evaluated using "Weibull" function parameters alpha and beta. Weibull parameters were used as dissolution markers of the eight principal, mainly compositional, variables. The parameters were obtained by fitting the Weibull function to the mean (n = 12) dissolution profiles of 28 diltiazem hydrochloride tablet formulations. The generated set of 28 pairs of Weibull function parameters was evaluated for internal and external predictability using both the MLR and the artificial NN. A three-layered 8-5-2 feedforward NN was found to be an adequate descriptor of the dissolution data. Internal predictions were based on the data of 24 products. External predictions used the 24 product data to test four products not used in the training phase. The predictive performances of the two techniques were evaluated using bias (mean prediction error; MPE) and precision (mean absolute error; MAE). The study results suggested that, for the studied data set, NN is a superior internal and external predictor to MLR. The artificial NN predicted order of the formulation composition variables, influencing the dissolution parameters as follows: hydrogenated oil > microcrystallinecellulose > ethyl cellulose > eudragit > hydroxypropylcellulose > coat > hydroxypropylmethylcellulose > Speed. PMID- 12741618 TI - Use of sodium chloride to facilitate reduction of particle size of dexamethasone during ball milling. PMID- 12741619 TI - Antiviral agents from plants and herbs: a systematic review. AB - BACKGROUND AND AIMS: Many antiviral compounds presently in clinical use have a narrow spectrum of activity, limited therapeutic usefulness and variable toxicity. There is also an emerging problem of resistant viral strains. This study was undertaken to examine the published literature on herbs and plants with antiviral activity, their laboratory evaluation in vitro and in vivo, and evidence of human clinical efficacy. METHODS: Independent literature searches were performed on MEDLINE, EMBASE, CISCOM, AMED and Cochrane Library for information on plants and herbs with antiviral activity. There was no restriction on the language of publication. Data from clinical trials of single herb preparations used to treat uncomplicated viral infections were extracted in a standardized, predefined manner. RESULTS: Many hundreds of herbal preparations with antiviral activity were identified and the results of one search presented as an example. Yet extracts from only 11 species met the inclusion criteria of this review and have been tested in clinical trials. They have been used in a total of 33 randomized, and a further eight nonrandomized, clinical trials. Fourteen of these trials described the use of Phyllanthus spp. for treatment of hepatitis B, seven reporting positive and seven reporting negative results. The other 10 herbal medicines had each been tested in between one and nine clinical trials. Only four of these 26 trials reported no benefit from the herbal product. CONCLUSIONS: Though most of the clinical trials located reported some benefits from use of antiviral herbal medicines, negative trials may not be published at all. There remains a need for larger, stringently designed, randomized clinical trials to provide conclusive evidence of their efficacy. PMID- 12741620 TI - Prevalence and clinical implications of insertions in the HIV-1 p6Gag N-terminal region in drug-naive individuals initiating antiretroviral therapy. AB - We assessed the prevalence and clinical impact of insertions within the HIV-1 p6Gag proline-rich (PTAP) region on initial antiretroviral therapy response in 461 HIV-infected, drug-naive individuals initiating therapy in British Columbia, Canada between June 1996 and August 1998. HIV p6Gag insertions were detected by nested RT-PCR of extracted patient plasma followed by direct DNA sequencing. Insertions were observed in 70 of 423 successfully genotyped samples (16.5%). HIV p6Gag insertions were significantly associated with a lower baseline CD4 cell count (P<<0.05) and the presence of basic amino acids at key positions in the HIV envelope V3 loop linked to a syncytium-inducing phenotype (P<<0.05). After adjusting for baseline factors, no effect of HIV p6Gag insertions was observed on time to virological success (pVL < or = 500 copies/ml), virological failure (subsequent confirmed pVL > or = 500 copies/ml) or immunological failure (confirmed CD4 count below baseline), as evaluated by Kaplan-Meier methods and Cox proportional hazard regression (P>0.1). The data suggest that HIV p6Gag insertions are not exclusively related to drug resistance and may not influence response to antiretroviral therapy, but may be linked to sequence variations in the HIV envelope. PMID- 12741621 TI - Shifts in cell-associated HIV-1 RNA but not in episomal HIV-1 DNA correlate with new cycles of HIV-1 infection in vivo. AB - The significance of distinct classes of HIV-1 nucleic acids as correlates of recent HIV-1 replication was assessed in peripheral blood mononuclear cells (PBMC) obtained from 14 patients during 2 weeks of structured interruption of antiretroviral therapy (STI) and 2 weeks of resuming therapy. Levels of HIV RNA in plasma (HIV-RNAplasma) and of unspliced cell-associated HIV-1 RNA (HIV UsRNAPBMC) were significantly increased as a result of STI, whereas no significant shifts in the levels of 2-LTR episomal HIV-1 DNA (2-LTR circles) and total late HIV-1 reverse transcripts (late-DNA) were observed. Thus, limited viral replication had occurred, which had no effect on the pool size of infected cells in the periphery. Levels of 2-LTR circles did not reflect rapid changes in HIV-1 replication. In contrast, expression of HIV-UsRNAPBMC increased during STI and consequently provides a more sensitive, albeit not absolute cellular marker of ongoing HIV-1 replication. PMID- 12741622 TI - ISDR pattern and evolution in patients with chronic hepatitis C treated with standard or PEG-IFN plus ribavirin. AB - The aim of the study was to characterize the interferon sensitivity determining region (ISDR) mutation pattern and its changes at 4 weeks of treatment in a population of patients infected with hepatitis C virus (HCV) genotype 1b receiving standard or PEG-IFN plus ribavirin (RBV), to find possible early correlates of therapy outcome. Forty-five patients with chronic hepatitis due to HCV 1b were treated by PEG-IFN-alpha2b (n=23) or IFN-alpha2b (n=22) plus RBV 1000 1200 mg/day. They were classified 24 weeks after stopping therapy as sustained responders (SR), relapsers (REL) or non-responders (NR). Sixteen patients were SR, 12 REL and 17 NR. ISDR mutations were evaluated by direct sequencing at baseline in all and after 4 weeks in patients with detectable viraemia (n=30). The frequency of the three ISDR types was 26.7% wild-type, 64.4% intermediate type and 8.9% mutant-type, without significant difference in their frequency in SR, REL and NR, independent of IFN formulation. Average numbers of mutations in SR, REL and NR were 1.88 +/- 0.54, 1.33 +/- 0.33 and 0.94 +/- 0.25, respectively, P>0.05. The baseline number of ISDR mutations was not related to the extent of viral load decline in the first month of therapy. Sequence analysis of ISDR region performed 4 weeks after starting therapy revealed qualitative or quantitative changes of ISDR sequence in only seven patients, without correlation with response. Thus, in our patients the baseline pattern of ISDR was unrelated to treatment outcome. Selection towards a dominant IFN-resistant strain did not occur under standard or PEG-IFN plus RBV. PMID- 12741623 TI - HIV-1 reverse transcriptase and protease resistance mutations selected during 16 72 weeks of therapy in isolates from antiretroviral therapy-experienced patients receiving abacavir/efavirenz/amprenavir in the CNA2007 study. AB - OBJECTIVE: To determine HIV-1 reverse transcriptase (RT) and protease (PRO) mutations selected in isolates from antiretroviral therapy (ART)-experienced patients receiving an efavirenz/abacavir/amprenavir salvage regimen. METHODS: Open-label, single arm of abacavir, 300 mg twice daily, amprenavir, 1200 mg twice daily and efavirenz, 600 mg once daily, in ART-experienced patients of which 42% were non-nucleoside reverse transcriptase inhibitor-naive. The virology population examined consisted of all patients who took at least 16 weeks of study drugs (n=74). Plasma population sequencing was carried out at baseline and last time point at which patients were still taking the three study drugs + other ART. The median follow-up was 48 weeks (range week 16-72). RESULTS: Baseline (n=73) and on-therapy (n=49) genotypes were obtained. By 48 weeks, 51% of isolates had > or = 3 non-nucleoside reverse transcriptase inhibitor (NNRTI) mutations. NNRTI mutations selected on therapy were K103N (51%), substitutions at position 190 (17/49, 35%): G to A (n=11) / S (n=4) / E (n=1) and T (n=1); L100I (37%) and V1081 (20%) mutations. P225H was not observed in this study. L100I and G190A/S/E/T mutations were rarely detected in the same viral population and baseline Y181C favoured the G190 mutations (OR=8.9, P<0.001), rather than the L100I. The NRTI mutations selected were in accordance with abacavir known resistance profile, no new TAMs were observed, new L74V or I mutations developed in 39 and 16% of isolates, respectively, however, new M184V mutations were only detected in isolates from two patients, one of whom had added lamivudine + didanosine. M184V was common at baseline (55%) and maintained in 22/27 (81%) isolates (five of these 22 added lamivudine or didanosine, or both). The PRO mutations selected were in accordance with the distinct resistance profile of amprenavir compared with other protease inhibitors. Mutations D30N, G48V, N88D/S, L90M and 154V were de-selected, and mutations I50V, I or V to 54M/L, I84V, M46I/L, L33F, I47V as well mutations at position 10 were observed in 20/49 (41%) isolates. CONCLUSION: Prior NNRTI and NRTI therapy influences the pathway of resistance to efavirenz. In this study, the prevalence of mutations selected by efavirenz were different from those described in less ART-experienced patients. Baseline Y181C was associated with the development of mutations at position 190, but not L100I or K103N. In this patient population, abacavir with efavirenz preferentially selected for L74V but not for thymidine analogue mutations. M184V was rarely selected and was maintained in only 77% of patients who did not add lamivudine or didanosine. Finally, amprenavir-specific mutations were selected in the background of other primary protease inhibitor mutations, confirming the distinct resistance profile of amprenavir. PMID- 12741624 TI - Salvage therapy with abacavir in HIV-1-infected patients with previously documented M184V mutation: a possibility of NRTI recycling. AB - We evaluated in an open-label, randomized, controlled, pilot trial if the re emergence of previously selected resistant strains, harbouring M184V mutation, could be modulated by the use of different drug associations as components of the new antiretroviral regimens. In addition, we assessed the clinical relevance of this mutation on the management of heavily pretreated HIV-infected patients. The primary end-point of the study was the reselection of M184V mutation. Secondary end-points were the variation over time of HIV RNA plasma levels and CD4 cell counts and the progression of HIV disease. The primary population for efficacy analysis was the intention-to-treat exposed population. After a run-in phase consisting in a new treatment regimen excluding either lamivudine (3TC) or abacavir (ABC) so as to clear the previously documented M184V mutation, 18 patients with an HIV RNA plasma level greater than 10000 copies/ml were randomized to receive an antiretroviral drug regimen (at least three drugs) including either ABC or the association of ABC+3TC. All patients were naive to ABC. The M184V mutation reappeared in 1/9 patients in the ABC group and in 8/9 patients in the ABC+3TC group (P<0.003, 95% CI: 0.5-1). In the ABC group we observed a rapid decrement of viral load that was maintained throughout all the study period (P<0.05). On the contrary, in the ABC+3TC group, after a transient decrement at 2 months, a progressive increment towards baseline values was observed. The proportion of patients with a viral load reduction of at least 0.5 logs at 12 months was significantly higher in the ABC group: 8/9 patients vs 3/9 (P=0.05, 95% CI: 0.2-0.92). Similarly, from an immunological point-of-view, the increase at all time points (since randomization) in CD4 cell count was statistically significant in the ABC group (P<0.01), while no difference was observed in the ABC+3TC group. The possibility of a successful use of ABC in salvage regimens opens alternative therapeutic options for heavily pretreated patients with previously documented M184V mutation. Further studies should clarify whether this is true for other drugs of the nucleoside analogues class. PMID- 12741625 TI - Changes in viral load in people with virological failure who remain on the same HAART regimen. AB - OBJECTIVES: To assess the rate of change in viral load and CD4 count over time in HIV-infected patients experiencing virological failure on a HAART regimen. DESIGN: Study population included patients from EuroSIDA, a large, multicentre, observational study enrolling HIV-infected patients across Europe. METHODS: Median change in viral load and CD4 count per month were estimated using the viral load and CD4 measurements obtained over a 12-month period after confirmed virological failure between 3 and 4 log10 copies/ml in a population of 488 HIV infected patients who were left on a failing HAART regimen. RESULTS: The estimated median viral load change in our study population was 0.024 log10 copies/ml per month, statistically different from 0 (P=0.0001). In 20.9% of the patients studied viral load showed a tendency to decrease, in 47.8% showed a tendency to increase by a positive rate no higher than 0.04 log10 copies/ml per month and in the remaining 31.3% showed a tendency to increase by a rate greater than 0.04 log10 copies/ml per month. On average, CD4 counts were estimated to remain stable (decrease at a slow rate of about -0.53 cells/microl per month). CONCLUSIONS: In patients that remained on a stable, but virologically failing HAART regimen (with viral load ranging 1000-10000 copies/ml), the viral load over the ensuing 12-month period increased at a relatively slow rate. In contrast, the CD4 count remained stable, possibly because of partial but sustained viral suppression below the viral load natural set-point. The time-course of selecting more replication-competent virus in patients with virological failure remains to be fully clarified. PMID- 12741626 TI - Dynamics of the pool of infected resting CD4 HLA-DR- T lymphocytes in patients who started a triple class five-drug antiretroviral regimen during primary HIV-1 infection. AB - Starting standard antiretroviral therapy within 10 days after the onset of a primary HIV-1 infection cannot prevent the establishment of a reservoir of HIV-1 infected memory CD4 T cells. Here we studied the reservoir of HIV-1-infected memory CD4 T cells in four patients who started a triple class, five-drug regimen during primary HIV-1 infection. There was a strong correlation between the proportion of productively infected CD4 HLA-DR- T lymphocytes and plasma HIV-1 RNA levels (r=0.852; P<0.001) during the first 24 weeks of therapy. Within 45 weeks of treatment, in three of the four patients the proportion of productively infected CD4 HLA-DR- T lymphocytes was reduced below the level of quantification. In the fourth patient the cellular reservoir remained quantifiable. In two patients who stopped therapy 44 weeks after initiation an immediate rebound of the plasma HIV-1 RNA level and the proportion of productively infected CD4 HLA-DR T lymphocytes occurred. In conclusion, initiation of a potent five-drug, triple class regimen during primary HIV-1 infection does not result in virus-specific immune control upon discontinuation of therapy after 44 weeks. Therefore, longer or even stronger suppression of viral replication might be necessary to achieve this goal in primary HIV-1 infection. PMID- 12741627 TI - Nucleotide analogue binding, catalysis and primer unblocking in the mechanisms of HIV-1 reverse transcriptase-mediated resistance to nucleoside analogues. AB - Nucleoside analogues play a key role in the fight against HIV-1. Unfortunately, under therapeutic pressure, HIV-1 inevitably develops resistance to these inhibitors. This resistance correlates with specific pol gene mutations giving rise to specific substitutions in reverse transcriptase that are responsible for the loss of efficacy of the corresponding analogue. This work is an overview of the molecular mechanisms of HIV-1 drug resistance as judged by the analysis of chemical reactions at play at the reverse transcriptase active site. One class of mechanism involves nucleotide analogue discrimination either at the binding step or at the catalytic step, the latter being by far the most common mechanism. The other class of mechanism involves repair of the analogue-terminated DNA chain. The mechanisms were elucidated using purified reverse transcriptase and biochemical assays aimed at correlating resistant HIV-1 phenotypes to enzymatic data. The elucidation of these molecular mechanisms of drug-resistant reverse transcriptase is important for effective and rational combination therapies as well as for the conception of second-generation drugs that do not confer nucleotide resistance to reverse transcriptase or are active against pre-existing resistant viruses. PMID- 12741628 TI - Anti-HIV-1 activity of enfuvirtide (T-20) by inhibition of bystander cell death. AB - Infection by human immunodeficiency virus type 1 (HIV-1) has been associated with increased cell death of both infected and bystander cells. The envelope glycoprotein complex appears to play an active role in HIV-induced death of bystander cells. We quantified cell-to-cell fusion, single cell death and membrane lipid mixing in cocultures of effector, HIV-1 envelope-expressing cells with peripheral blood mononuclear cells or purified CD4 T lymphocytes from HIV negative donors, in the presence or the absence of the fusion inhibitor enfuvirtide (T-20, pentafuside, Fuzeon). T-20, which blocks gp41-dependent virus cell fusion, showed a complete and dose-dependent inhibition of syncytium formation in cocultures of envelope-expressing cells with uninfected cells. Similarly, T-20 totally abrogated death of single bystander CD4 T cells with an IC50 of 0.04 microg/ml. Membrane lipid mixing, as a measure of interaction between envelope-expressing cells and CD4 cells, was also dose-dependently inhibited by T-20. Moreover, effector cells chronically infected with a T-20 resistant virus recovered the ability to induce bystander cell death in the presence of the drug, supporting the role of gp41 in single cell death. In conclusion, T-20 is able to protect CD4 T cells from envelope presentation with a dual effect: inhibition of virus replication and blockade of HIV-1 envelope induced cell death of bystander CD4 T cells. Protection of cells prior to infection from HIV envelope-dependent bystander effect could lead to a better immune restoration of HIV-1-infected patients that are treated with T-20. PMID- 12741629 TI - Triple nucleoside combination zidovudine/lamivudine/abacavir versus zidovudine/lamivudine/nelfinavir as first-line therapy in HIV-1-infected adults: a randomized trial. AB - OBJECTIVE: To compare the efficacy and safety of a triple nucleoside combination to a protease inhibitor-containing triple regimen as first-line antiretroviral therapy (ART) in HIV-1-infected patients. DESIGN: Open-label study in HIV-1 infected ART-naive adults, randomized to receive either Combivir (lamivudine 150 mg/zidovudine 300 mg twice daily) + abacavir (300 mg twice daily), or Combivir + nelfinavir (750 mg every 8 h) for 48 weeks. Plasma HIV-1 RNA, CD4 cell count and adverse events were assessed at baseline and weeks 4, 8, 16, 24, 32, 40 and 48. RESULTS: 195 subjects (131 men, 64 women), median age 34 years, were randomized: 98 received combivir/abacavir and 97 combivir/nelfinavir. Baseline median plasma HIV-1 RNA was 4.2 log10 copies/ml [Interquartile range (IQR): 3.7-4.5.2] and 4.1 log10 copies/ml (IQR: 3.8-4.6), respectively. Baseline median CD4 cell count was 387 cells/mm3 (IQR: 194-501) and 449 cells/mm3 (IQR: 334-605), respectively. Nine patients (3 vs 6, respectively) did not start treatment or did not have any available efficacy data. At week 48, using the intent to treat analysis (switch/missing equals failure), plasma HIV-1 RNA was <50 copies/ml in 54/95 (57%) and 53/91 (58%) of subjects, respectively. Median CD4 increase was +110 and +120 cells/mm3, respectively. Possible hypersensitivity reactions to abacavir were reported in four subjects (4%). CONCLUSION: The triple nucleoside combination combivir/abacavir is well tolerated as a first-line ART regimen in HIV-1-infected adults, with comparable antiviral activity to a nelfinavir containing regimen at week 48. PMID- 12741631 TI - In profile: Peter D. Wentzell, Dalhousie University. PMID- 12741630 TI - K65R with and without S68: a new resistance profile in vivo detected in most patients failing abacavir, didanosine and stavudine. AB - Antiretroviral treatment with three nucleoside reverse transcriptase inhibitors (NRTIs) is widely used, but the combination of abacavir, didanosine and stavudine has never been investigated. We describe the surprising and consistent genotypic and phenotypic outcome in patients failing this combination. As part of a Danish multicentre study, 60 antiretroviral-naive patients were randomized to treatment with abacavir, didanosine and stavudine. Failure was defined as one HIV-1 RNA >400 copies/ml. Genotyping was performed using TrueGene HIV-1 assay (Visible Genetics, London, UK). Phenotypic susceptibilities were determined with the Virco Antivirogram assay. Eight patients failed treatment with a median viral load of 2.980 copies/ml (range 478-5.950). At baseline, five patients were wild-type. Three patients harboured nucleoside excision mutations (NEMs), but phenotypic susceptibilities were within normal range. All five patients with wild-type virus developed K65R and four of these patients also acquired the S68G mutation. Phenotypic susceptibility decreased towards abacavir (median 8.9-fold) and didanosine (median 3.2-fold), while susceptibility towards stavudine was unchanged (median 0.8-fold). Susceptibility towards lamivudine and tenofovir decreased median 14.2- and 4.0-fold, respectively. In two patients with baseline resistance mutations, further accumulation of NEMs and V75T or L74V was observed. One patient developed Q151M. Failure of a triple NRTI regimen is possible and frequent with only the K65R mutation. Under adequate selection pressure K65R can easily emerge in vivo and may compromise several future treatment options including newer NRTIs. The unexpected high incidence of S68G suggests a functional role of this mutation in viruses harbouring K65R. PMID- 12741632 TI - Optical nanosensors--an enabling technology for intracellular measurements. AB - Optical nanosensors have been designed to utilise the sensitivity of fluorescence for making quantitative measurements in the intracellular environment, using devices that are small enough to be inserted into living cells with a minimum of physical perturbation. Advantages over widely used fluorescence dye based methods are observed because the nanosensor matrix imparts two key benefits; (1) protection of the sensing component from interfering species within the intracellular environment and (2) protection of the intracellular environment from any toxic effects of the sensing component. This Highlight article discusses the recent developments in nanosensor technology and investigates the use of more complex sensing schemes that will expand the range of analytes that can be detected and quantified. PMID- 12741633 TI - Fluorescence polarization spectroscopy in protein analysis. AB - Information about concentration, molecular structure, binding events, and motion can be obtained using fluorescence spectroscopy methods. Fluorescence polarization spectroscopy is one such method, which studies the relationship between the polarization of light that is used for excitation and light that is subsequently detected from fluorescence. The extent of change of polarization between excitation and emission can be used to study physical processes such as rotational diffusion, extent of denaturation, and orientation at surfaces. In this article Mann and Krull describe the underlying principles to the technique and show how fluorescence polarization spectroscopy has contributed to protein analysis. PMID- 12741634 TI - Bioscience and analytical thinking machines. AB - Is science being limited by the restricted vision of the analytical scientist? Adrian Stevenson from the Institute of Biotechnology at University of Cambridge argues that human 3D visualisation abilities to find patterns are inadequate and that artificial intelligence may hold the vital key to understanding information dense biological systems. PMID- 12741635 TI - The Walkerton tragedy --issues for water quality monitoring. AB - One of the most significant weaknesses in modern day water quality management is the detection of microbiological indicators. Microbial tests are normally conducted off-site, often resulting in long turn-around time, risk of contamination, cross contamination and adulteration. Here Stephen Brown and Moe Hussain present a thought provoking case study where limitations in current analytical technologies for water monitoring had fatal results and discuss new approaches to microbiological monitoring that might prevent similar disasters occurring in the future. PMID- 12741636 TI - Stimulus-sensitive hydrogels and their applications in chemical (micro)analysis. AB - In this tutorial review the use of stimulus-sensitive hydrogels as sensors and actuators for (micro)analytical applications is discussed. The first part of the article is aimed at making the reader familiar with stimulus-sensitive hydrogels, their chemical composition and their chemo-physical behavior. The prior art in the field, that comprises a number of sensors ranging from metal ion-sensitive sensors to antigen-sensitive sensors and a few actuators, is also treated in this part. The second part of the article focusses on the use of stimulus-sensitive hydrogels for microsensors and microactuators as well as their application in micro total analysis systems. The benefits of stimulus-sensitive hydrogels, their miniaturisation and the use of 365 nm UV-photolithography as a fast economical manufacturing technique are discussed. PMID- 12741637 TI - Soxhlet extraction of acrylamide from potato chips. AB - The problem of complete extraction of acrylamide from potato chips was investigated. A method was developed based on the Soxhlet extraction technique. A defatted sample was extracted continuously with methanol, for 10 days, in a Soxhlet extractor. After about 7 days, a constant concentration of acrylamide was reached. This indicates that all the acrylamide that could be removed from the sample had been extracted. Acrylamide was identified in the extract using GC-MS and scan mode. Total concentration was 14500 microg kg(-1) using GC-FID and standard additions. Complementary determinations, using an external standard (GC FID and GC-MS) and an internal standard (GC-FID), showed results within +/- 5%. A previously published study, using a static extraction method and GC-MS and LC-MS MS, showed concentrations of 2287 and 1993 microg kg(-1), respectively. The results are discussed in relation to a recent model and analogous experiments. The extracted amount of acrylamide is affected by several parameters: solvent properties, solvent volume, extraction time, temperature, particle size, and the microstructure of the sample. PMID- 12741638 TI - Non-trivial temperature effects on the cation exchange chromatography and chelation ion chromatography of metal ions. AB - The effect of column temperature upon the retention of metal ions on sulfonated and mono-, di-, and amino-carboxylated cation exchange columns has been investigated. The retention of alkali, alkaline earth and transition metal ions on each of the above types of cation exchanger was studied over the temperature range 19-65 degrees C. A major difference between the behaviour of mono- and divalent metal ions was shown on each of the above stationary phases, with the monovalent alkali metals exhibiting clearly exothermic behaviour (a decrease in retention with increased temperature) under acidic eluent conditions and an apparent relationship between retention factor and the magnitude of the temperature effect. The effect of temperature upon alkaline earth metal ions was less defined, although strongly endothermic behaviour (increase in retention with temperature) could be seen on all stationary phases through correct choice of eluent. The transition metal ions studied showed endothermic behaviour on all four stationary phases, with the sulfonated column unexpectedly showing the largest increases in retention. The above behaviour can be partially explained through the dominance of the type of solute-stationary phase interaction governing retention. In several of the above columns, both ion-exchange and surface complexation interactions can occur, with the effects of temperature indicating which process dominates under specific eluent conditions. PMID- 12741639 TI - Comparison of normal and reversed-phase solid phase extraction methods for extraction of beta-blockers from plasma using molecularly imprinted polymers. AB - A molecularly imprinted polymer (MIP) prepared using propranolol as template, methacrylic acid (MA) and ethylene glycol dimethacrylate (EGDMA) was used to develop SPE methods in "reversed-" and normal phase mode for an analogue of propranolol (M47070) with another analogue (M45655) used as an internal standard. The compounds were also extracted in reversed-phase mode onto a non-imprinted polymer. It was necessary to employ a protein precipitation step ahead of MIP-SPE in order to facilitate downstream analysis. High extraction efficiencies and linear calibration ranges were achieved using both reversed-phase (RP) and normal phase (NP) MIP-based methods. Extraction efficiencies were lower on the non imprinted polymer indicating stronger retention by the MIP. This stronger retention was attributed to selective imprint-based binding by the MIP that was not available for the non-imprinted polymer. Although clean extracts were obtained in both RP and NP modes, low level interference from template-related impurities or degradation products compromised detection of M47070 at low concentrations for the MIP-based methods. This interference made accuracy of the MIP-based methods poorer at low concentrations. The reversed-phase method showed marginally better accuracy and precision than the normal phase method. PMID- 12741640 TI - On-line coupling of sequential injection extraction with restricted-access materials for sample clean-up and analysis of drugs in biological matrix. AB - In this contribution, the on-line coupling of solid phase extraction (SPE), based on a restricted-access material (RAM), with sequential injection technique (SIA) for the analysis of biological samples, is described. The SIA-RAM system was tested with a new potential antileucotrienic drug (VUFB-19363 (Quinlukast)) for serum analysis. The method is based on SPE with the novel internal-surface reversed-phase column packing material-alkyl-diol silica (ADS). The supports tolerate direct and repetitive injection of proteinaceous fluids (plasma, serum) and allow reversed-phase partitioning at the internal surface. A column packed with a 25 microm C18 alkyl-diol support was used for direct serum injection. Using a 6-port selection valve and the system of three mobile phases, the polar matrix compounds and metabolites are removed by sequentially aspirated mobile phases with lower content of the organic part (methanol-water (2:98) and following acetonitrile-water (20:80)) to the waste, and then, the analyte enriched on the column is eluted by a strong mobile phase (acetonitrile-methanol water (40:20:40)) to the UV detector without transfer loss. With the fully automated SIA system, a total analysis time of less than 10 min was achieved. The only off-line sample pre-treatment step required to remove particulate matter was centrifugation. The studies showed a range of linearity (2-40 microg ml(-1)) and a high recovery (93.6-96.8%) of drug from the biological matrix with coefficients of variation (RSD) less than 5.0% (n = 6). This paper introduces a new, simple and robust analytical technique suitable for screening determination and direct analysis of drugs in biological materials. PMID- 12741641 TI - Solid-phase immunoassay detection of peptides from complex matrices without a separation. AB - A simple and sensitive solid-phase fluorescence immunoassay method was developed to detect peptides without separating them from a biological matrix. A near infrared fluorescence detection system was constructed for scanning analyte spots blotted onto protein binding membranes. Hydrophobic membranes were used with a modified vacuum spot blotting system to concentrate the peptide solution into a small area and the overall assay time was thus reduced by eliminating blocking steps. Both direct and indirect immunoassay methods are demonstrated; the indirect is more sensitive and features a 1 pmol detection limit of neat dynorphin A solutions. To further increase the immunoassay sensitivity, a novel capillary blotting system with hydrophilic membranes was designed where optimized sample volumes of 167 nL were deposited for each spot. The area-reduced blotting method shows a 1000-fold improved, 1.3 fmol spot(-1) detection limit of a dynorphin A diluted in a buffered solution of 150 mg L(-1) of casein. Low-flow push-pull perfusates with volumes of 1 microL sampled from the striatum of the rat were assayed for dynorphin A by the method of standard addition. The detection limit was estimated to be 1.9 fmol in the low-flow push-pull perfusates. These data demonstrate a solid-phase near infrared immunofluorescence strategy for the study of peptides directly blotted from chemically complex biological fluid matrices. PMID- 12741642 TI - Quantitative measurement of male steroid hormones using automated on-line solid phase extraction-liquid chromatography-tandem mass spectrometry and comparison with radioimmunoassay. AB - A specific and sensitive method using high-performance liquid chromatography tandem mass spectrometry (LC-MS-MS) equipped with automatic on-line solid-phase extraction device for the quantitative measurement of anabolic hormone residues, 4-androstene-3,17-dione, testosterone and dihydrotestosterone in cell culture medium was developed. Steroid content in cell culture medium was determined directly without an additional sample preparation step. Separation of analytes from polar endogenous compounds was carried out on an automatic column-switching device coupled with a C4-alkyl-diol silica restricted-access solid-phase extraction column. The lipophilic fraction containing anabolic hormone residues were back-flushed on to a conventional C-18 reversed-phase column for the final chromatography. The analyte was ionized in an ElectroSpray interface under positive ion mode before entering a quadrupole mass analyzer. The lowest points of calibration curves were 0.05 ng ml(-1) for 4-androstene-3,17-dione and testosterone, and 1 ng ml(-1) dihydrotestosterone, respectively. A comparison with results from radioimmunoassay (RIA) is also presented. PMID- 12741643 TI - Sensitive determination of ephedrine and pseudoephedrine by capillary electrophoresis with laser-induced fluorescence detection. AB - A CE-LIF method was developed for the separation and sensitive detection of ephedrine and pseudoephedrine after derivatization by 4-chloro-7-nitrobenzo-2-oxa 1,3-diazol (NBD-C1). The derivatization and separation conditions were investigated in detail and the optimum conditions were obtained. Under the optimum experiment conditions, good linearity relationships (correlation coefficients: 0.9942 for ephedrine and 0.9970 for pseudoephedrine) between the peak heights and concentrations of the analytes were obtained (0.7-140 microM). The detection limits were 0.16 microM for ephedrine and 0.17 microM for pseudoephedrine, which indicated that the sensitivities were at least ten times improved over those reported in the literature obtained by UV detection. The method was applied to the analysis of ephedrine and pseudoephedrine in ephedra herb plants and preparations with good results. PMID- 12741644 TI - Effect of non-significant proportional bias in the final measurement uncertainty. AB - The trueness of an analytical method can be assessed by calculating the proportional bias of the method in terms of apparent recovery. If the apparent recovery does not differ significantly from one, the analytical method has not a significant bias. If this is the case, the bias is neglected and the uncertainty associated with this bias is included in the uncertainty budget of results. However, when assessing trueness there is always a probability of incorrectly concluding that the proportional bias is not significant. Therefore, the uncertainty of results may be underestimated. In this paper, we study how non significant bias affects the uncertainty of analytical results. Moreover, we study how to avoid the underestimation of uncertainty by including the non significant bias calculated in the uncertainty budget. To answer these questions, we have used the Monte-Carlo method to simulate the process of estimating the apparent recovery of a biased analytical method and, subsequently, the future results this method provides. The results of the simulation show that non significant bias may underestimate the uncertainty of analytical results when bias contributes in more than 20% to the overall uncertainty. Uncertainty is specially underestimated when bias contributes in more than 50% to the overall uncertainty. PMID- 12741645 TI - Multi-analyte optimisation of uncertainty in infant food analysis. AB - The Optimised Uncertainty (OU) methodology has been developed to optimise multi analyte situations. It has then been applied to a retail survey of infant food for trace elements, classifying the food as compliant or non-compliant with the regulatory thresholds or specification limits that are appropriate for each element. The large-scale survey of infant foods was successfully adapted to allow the estimation of uncertainties, from both primary sampling and chemical analysis, for elemental concentrations in infant formula (milk) and wet meals. The analytes included in this investigation comprised both contaminants (Pb and Cd) and elements essential for child development (Zn and Cu). Optimisation of the measurement process for a 'single analyte' demonstrated the potential financial benefits of optimising future surveys for a false compliance scenario. Uncertainty estimates for the measurement of elemental concentrations in infant formula were dominated by uncertainty from the analytical method. Large potential savings (up to pounds 575,000 per batch) are predicted for both Pb and Zn by increasing the expenditure on chemical analysis to the optimal level. In comparison the uncertainty estimates for elemental concentration in wet meals showed a dominance of sampling as a source of uncertainty for Cd and Cu due to the increased heterogeneity. The feasibility of 'multi-analyte' optimisation is demonstrated for the case study of infant milk. Single analyte optimisation of the four analytes for a false compliance scenario indicated a decrease in expectations of financial loss of between 99% and 8%. An overall decrease in the total expectation of financial loss of 99% is indicated following multi-analyte optimisation. PMID- 12741646 TI - NIR spectroscopic measurement of moisture content in Scots pine seeds. AB - When tree seeds are used for seedling production it is important that they are of high quality in order to be viable. One of the factors influencing viability is moisture content and an ideal quality control system should be able to measure this factor quickly for each seed. Seed moisture content within the range 3-34% was determined by near-infrared (NIR) spectroscopy on Scots pine (Pinus sylvestris L.) single seeds and on bulk seed samples consisting of 40-50 seeds. The models for predicting water content from the spectra were made by partial least squares (PLS) and ordinary least squares (OLS) regression. Different conditions were simulated involving both using less wavelengths and going from samples to single seeds. Reflectance and transmission measurements were used. Different spectral pretreatment methods were tested on the spectra. Including bias, the lowest prediction errors for PLS models based on reflectance within 780 2280 nm from bulk samples and single seeds were 0.8% and 1.9%, respectively. Reduction of the single seed reflectance spectrum to 850-1048 nm gave higher biases and prediction errors in the test set. In transmission (850-1048 nm) the prediction error was 2.7% for single seeds. OLS models based on simulated 4 sensor single seed system consisting of optical filters with Gaussian transmission indicated more than 3.4% error in prediction. A practical F-test based on test sets to differentiate models is introduced. PMID- 12741647 TI - The automated sample preparation system MixMaster for investigation of volatile organic compounds with mid-infrared evanescent wave spectroscopy. AB - For efficient development assessment, and calibration of new chemical analyzers a large number of independently prepared samples of target analytes is necessary. Whereas mixing units for gas analysis are readily available, there is a lack of instrumentation for accurate preparation of liquid samples containing volatile organic compounds (VOCs). Manual preparation of liquid samples containing VOCs at trace concentration levels is a particularly challenging and time consuming task. Furthermore, regularly scheduled calibration of sensors and analyzer systems demands for computer controlled automated sample preparation systems. In this paper we present a novel liquid mixing device enabling extensive measurement series with focus on volatile organic compounds, facilitating analysis of water polluted by traces of volatile hydrocarbons. After discussing the mixing system and control software, first results obtained by coupling with an FT-IR spectrometer are reported. Properties of the mixing system are assessed by mid infrared attenuated total reflection (ATR) spectroscopy of methanol-acetone mixtures and by investigation of multicomponent samples containing volatile hydrocarbons such as 1,2,4-trichlorobenzene and tetrachloroethylene. Obtained ATR spectra are evaluated by principal component regression (PCR) algorithms. It is demonstrated that the presented sample mixing device provides reliable multicomponent mixtures with sufficient accuracy and reproducibility at trace concentration levels. PMID- 12741649 TI - Determination of the amino acid tryptophan and the biogenic amine tryptamine in foods by the heavy atom induced-room temperature phosphorescence methodology. AB - Very simple and selective methods are presented to determine the amino acid tryptophan (Trp) and the biogenic amine tryptamine (Tryp), both compounds with an indole-type molecular structure by the methodology named Heavy Atom Induced-Room Temperature Phosphorescence (HAI-RTP) which constitutes the first time that HAI RTP has been used to detect compounds with non-naphthalenic structures in their molecules. Different variables affecting the phosphorescence signal (heavy atom perturber and sodium sulfite concentration) were carefully studied. The analytical curves give a linear dynamic range of 15-100 ng ml(-1) and a detection limit of 4 ng ml(-1) for Trp and 94-400 ng ml(-1) and 28 ng ml(-1) for Tryp. The methods have been successfully applied to the analysis of complex food matrices such as the presence of tryptophan in yoghurt and tryptamine in bottled beer. A single alkaline hydrolysis to release Trp from yoghurt proteins and two methods for extracting Tryp from beer samples are proposed and optimised. A total Trp content of 374 mg of Trp per kg of yoghurt was quantified by the standard addition method of calibration and a recovery of 90% was obtained for 250 ng ml( 1) of Tryp in spiked non-alcoholic beer samples. PMID- 12741648 TI - Solid-phase fluoroimmunoassay for the determination of mesotrione--a novel triketone herbicide--in water with direct measurement of the fluorescence onto the solid support. AB - A straightforward, low-cost fluoroimmunoassay (FIA) for the determination of the new triketone herbicide mesotrione has been developed and optimized. The protein mesotrione conjugate, immobilized on white opaque microtitration wells competes with the mesotrione in the sample or standard for the limited binding sites of a liquid phase anti-mesotrione antibody. The assay is based on the measurement of fluorescence intensity directly onto the solid support, using a fluorescein labeled second antibody and a fluorescence plate reader. To stabilize and enhance the fluorescence signal a glycerine-based treatment of the microtitration wells was included in the protocol. The detection limit of the assay is 40 ng 1(-1) (4 pg per well), the working range extends up to 9 microg l(-1), whereas the within and between run CVs are 0.7-4.2% and 2.1-5.5%, respectively. To evaluate the assay specificity, the cross-reactivities of two mesotrione metabolites: 4 methylsulfonyl-2-nitrobenzoic acid and 2-amino-4-methylsulfonyl-benzoic acid and several other compounds similar in structure to mesotrione such as: fomesafen, prosulfocarb, fluazinam, sulcotrione, 1,2-cyclohexanedione, 1,3-cyclohexanedione, 2-acetyl-1,3-cyclohexanedione were assessed. Most of the substances tested presented very low ( < 0.05%) cross-reactivity values with the exception of sulcotrione that cross-reacted 23% in the mesotrione assay. The assay was used for the determination of mesotrione in bottled natural waters fortified with the analyte and in a commercial herbicide formulation, namely CALLISTO. PMID- 12741651 TI - Long-term psychiatric morbidity in the aftermath of CS spray trauma. AB - The physical effects of CS spray are well documented. However, less is known about the impact of its use by the police on the psychological health of those exposed. In this study we explored the psychiatric morbidity of a group of 30 individuals who experienced the same trauma, a significant part of which was exposure to CS spray. Just over a quarter suffered post-traumatic stress disorder. It is proposed that more attention needs to be paid to the psychological effects of the use of CS spray by the police. Consistent with other studies, a past psychiatric history and a more external locus of control was associated with post-traumatic morbidity. The latter finding may have implications for psychological interventions after exposure to trauma. PMID- 12741652 TI - Multiple contacts with diversion at the point of arrest. AB - A diversion at the point of arrest (DAPA) scheme was set up in five police stations in South Birmingham in 1992. In a study of all referrals made over a four-year period a sub group of multiple contact individuals was identified. During that time four hundred and ninety-two contacts were recorded in total, of which 130 were made by 58 individuals. The latter group was generally no different from the single contact group but did have a tendency to be younger. This research highlights the need for a re-evaluation of service provision and associated education of police officers and relevant mental health care professionals. PMID- 12741650 TI - The role of the law in the care of sick children. PMID- 12741653 TI - Astrological birth signs in suicide: hypothesis or speculation? AB - Astrology is no longer regarded as a science by many, because its claims are almost impossible to test empirically in controlled laboratory conditions and it can not meet the scientific need to be reproducible. However, the majority of those who read their 'star signs' can identify aspects of their personality in what they read and it is possible that this may influence their attitudes and actions. The literature has neglected astrological signs as a possible predictor of suicide ideation. To see whether astrological birth signs are associated with suicide and the method used, data was collected from the Public Health Department in North Cheshire representing all the Cheshire Coroner's verdicts of suicide, and open verdicts, in all deceased aged 60 and above between 1989 and 2000. The observed occurrence of deaths due to natural causes, and suicide, in relation to birth signs did not differ significantly from what would be expected from chance. However, the distribution of suicide by hanging appeared significantly higher in those with a birth sign of Virgo and lowest in Sagittarius and Scorpio. The distribution of violent and non-violent suicides in relation to star signs showed higher occurences of violent death in persons born in the summer months. PMID- 12741654 TI - Cannabis and violent crime. AB - We present a series of 12 cases of violent crime, which were all committed under the influence of cannabis in Geneva, Switzerland, between 1996 and 2000. The crimes were committed by eleven males and one female, with a mean age of 26 years, who were using only cannabis at the time they acted. Most of them were chronic users. Five subjects had a past psychiatric history. Five had a personality disorder. Only three had been sentenced in the past for violent acts. At the time of the aggression, all of them exhibited adverse and acute effects of cannabis. All of them were judged by the court to be partially or totally non responsible. Three cases are presented in more detail. Our data suggests that cannabis could have a specific role in the development of violent behaviour patterns and that detection of its adverse effects should be systematic in criminal responsibility evaluation. PMID- 12741655 TI - Psychiatric treatment in prison: a missed opportunity? AB - National Health Service Executive guidelines require psychiatric services to maintain links with prisoners previously subject to the Care Programme Approach (CPA) and to participate in discharge planning. We are unaware of previous studies assessing the involvement of general psychiatric services with patients in prison or prisoners' perceptions of their needs. Consecutive referrals to a prison psychiatric liaison service over a three-month period were screened for previous psychiatric contact. Half of those interviewed reported previous psychiatric contact. Two-thirds were in contact with services at the time of detention. One-third believed services knew of their imprisonment. Ninety-three per cent believed they would require psychiatric support after release. Few patients received input from general psychiatric services during imprisonment despite a high level of perceived need. Improved liaison would help facilitate both care in prison and discharge planning in the spirit of CPA and the government directive. PMID- 12741656 TI - Accidental fatal hypothermia in elderly people with Alzheimer's disease. AB - We report two forensic autopsy cases of fatal accidental hypothermia in an 89 year-old woman and a 76-year-old man who were found dead and unclothed. In both cases, Alzheimer's disease (AD) was diagnosed by neuropathological examination. Wandering due to AD was determined as the cause of these accidents. Although paradoxical undressing in hypothermic victims is known to occur as a result of cold exposure, in our patients, undressing was attributed to dementia due to AD before they became hypothermic. These cases indicate that neuropathological examination is crucial to determining the cause of such accidents and that undressing is not always the result of hypothermia in elderly victims. PMID- 12741657 TI - The use of consent-to-treatment forms at the state hospital: an audit in 1996 and 2000. PMID- 12741658 TI - Psychiatrists' knowledge of the Human Rights Act and its relevance to mental health practice: a questionnaire survey. AB - The recent implementation of the Human Rights Act 1998 has important implications for UK psychiatric practice but previous studies have demonstrated that psychiatrists generally have poor knowledge of mental health law. The aims of this study were to assess psychiatrists' knowledge of the Human Rights Act and to examine whether knowledge is related to seniority or experience. Questionnaires were sent to 154 psychiatrists in the Northern Region, testing both their factual knowledge of the Human Rights Act and their ability to apply it to clinical scenarios. Ninety-mix psychiatrists responded and they demonstrated good overall knowledge and ability to apply the Human Rights Act However, half of the respondents were not aware that the Act only imposes a duty on public authorities and that a lack of active treatment in itself does not constitute a breach of the Act Specialist registrars and consultants scored significantly higher than SHOs but there was no significant difference between the specialist registrars and consultants. The scores of psychiatrists who had detained at least one patient under the Mental Health Act 1983 over the last six months were significantly higher than those who had not. The implications of these findings are discussed. There is still a need for further training on relevant aspects of the Human Rights Act. PMID- 12741659 TI - Suicide trends in Singapore: two decades down the road. AB - In this study, recent trends in the incidence and methods of suicide in Singapore, over the period 1991-2000, were compared with the results of a previous study covering the period 1975-1984 (Tan, 1986). The present study included a total of 3,834 suicidal deaths reported to the coroner and investigated by the Centre for Forensic Medicine of the Health Sciences Authority. The results showed that the crude suicide rate had stabilised over the last decade and that the rate was highest among elderly males. The three most common methods employed were falls from a height (69.3%), hanging (20.7%) and poisoning (5.5%). Comparison of the results of both studies showed that the crude suicide rate had stabilised over the last two decades. However, there was an increase in the suicide rates among males, as compared with the previous study, and a marginal decrease in suicide rates among females over this time. There was also an appreciable change in the methods of suicide employed, in that there was an increase in the proportion of deaths due to falls from a height and corresponding reductions in the proportions of deaths by hanging and poisoning. PMID- 12741660 TI - Women's rights in Pakistan: a forensic perspective. AB - Pakistan is a large and an important West Asian country which came into being in the name of Islam and therefore Islamic tenets remain the core of its constitution. The laws of the state have to conform to Islamic law so that they can have a positive impact on the society. Unfortunately, in Pakistan today not all men enjoy the rights and facilities to which they are entitled and women are doubly disadvantaged by poverty and gender. With their own political agendas, various governments have promulgated laws which affect the society in various ways. The laws which directly influence women's rights merit mention, as women comprise more than 50% of the population of Pakistan and are still kept on the sidelines by the male dominant society. The Muslim Family Law Ordinance, 1961, and the Hudood Ordinance, 1979 were both promulgated by military dictators with different visions. The former codified the rights of women bestowed by Islamic law; the latter repealed laws for sexual offences according to the injunctions of Islam and had a negative impact. Both laws need the assistance of forensic medicine as age estimation and medical examinations are necessary if they are to be followed in the right perspective. However, a legal need for an examination by an expert in forensic medicine is sadly lacking in both laws. This has happened due to lack of training of forensic physicians and therefore a lack of research in important areas of forensic medicine in the country. This paper examines these laws and the interaction they have with forensic medicine and proposes that the laws need revision in accordance with modern science, incorporating forensic sciences as well as the injunctions of Islam. PMID- 12741661 TI - Child homicide. AB - Between 1967 and 1988, 69 cases of single perpetrator/single victim child homicide resulted in remands into custody in the Yorkshire region. Sixty-four of these cases were examined retrospectively to identify the characteristics of the perpetrators and of victims under 16 years, the relationship of the victim to the accused and the circumstances of the offence. Sixty-four men singly accused of killing a single child victim are described in detail. They were characterized by relatively young age and a lack of long-term stable relationships. Previous psychiatric contact and/or a history of self-harm was noted in one-third of cases. Over half of the group had a criminal record and previous violence to children was noted in 28% of cases. Fathers or surrogate fathers accounted for nearly two-thirds of the accused. In terms of the victims, children under six months were at greatest risk. Nearly one-third of victims were the biological offspring of the accused. Sexually motivated homicide accounted for approximately 18.7% of deaths. Victim behaviours and domestic disharmony acted as precipitants in 64% of the cases, with 54.7% of the victims dying as a result of physical beatings. Alcohol consumption at the material time was more common than noted in previous studies of child homicide. PMID- 12741662 TI - The Mental Health Act 1983 section 5(2): clinical application and predictors of outcome. AB - This study examined trends and influences of variables in the 100 most recent applications of Section 5(2) of the Mental Health Act 1983 in a major teaching hospital in England. Case notes, section-forms, log-books and computerised records were scrutinised. Sixty-four per cent of the cases were converted to Section 2(28%) or Section 3(36%), the remainder we rescinded or lapsed. No demographic, illness characteristic or grade of doctor significantly effected conversion or lapse rate of Section 5(2). The clinical implications of a high rate of non-conversion of Section (2) is clear. Non-psychotic patients may be less likely to be regarded for further Sections when detained under Section 5(4) before detention under Section 5(2), yet no ingle factor determines the outcome of emergency. PMID- 12741663 TI - Traumatic basal subarachnoid haemorrhage caused by the impact of a golf ball: a case report. AB - A 50-year-old male was hit by a high-speed golf ball on the left lateral side of his neck. He collapsed immediately and was sent to hospital, where he was pronounced dead. The autopsy showed an extensive basal subarachnoid haemorrhage. Careful gross and histological examinations disclosed a rupture of the right vertebral artery at a site very close to the bifurcation. It was estimated that the impact of the golf ball on the left side of his neck resulted in the rupture of the contralateral vertebral artery, owing to its hyperextension. Although there are many reports on traumatic basal subarachnoid haemorrhage, the present type of trauma seems rare to our knowledge. PMID- 12741664 TI - An unusual suicide by stabbing: a case report. AB - A 50-year old cobbler, employed in a paramilitary unit, was found dead in a pool of blood in his shop inside the unit line in the morning. He had multiple stab wounds on his chest and was alleged to have committed suicide by repeated self stabbing on the chest with a sharp iron chisel, which was found clenched in his right hand by the investigating officer. Fifteen wounds were communicating with the chest cavity, injuring the vital anatomical structures. The individual, according to the unit personnel, was not suffering from any physical or mental disorder and was not on any medication. PMID- 12741665 TI - Abstracting information. PMID- 12741666 TI - The bereavement experience following home-based family caregiving for persons with advanced cancer. AB - The purpose of this qualitative interpretive study was to explore the experience of bereavement following home-based family caregiving for persons with advanced cancer. The research question addressed by this research was: How do family caregivers of patients with advanced cancer perceive the effects of home-based caregiving on their bereavement? Fifteen caregivers were retrospectively interviewed twice after the death of their family member. Caregivers reported both positive (e.g., feelings of accomplishment, improved family relationships) and negative (e.g., haunting images, feelings of failure) outcomes that they attributed to having cared for their family member. Overall positive outcomes predominated and bereaved family members reported satisfaction with having provided care for their loved one who had died. PMID- 12741667 TI - Clinical profile of acute confusion in the long-term care setting. AB - Aspects of acute confusion (AC) including risk factors, behavior patterns, and outcomes are not well documented in long-term care (LTC) residents. The purpose of this prospective study was to describe the clinical profile of AC in LTC including risk factors, behavior patterns, etiologies, and 3-month outcomes. Seventy-four elderly LTC residents were assessed for AC, depression, and global cognitive impairment. Risk factors associated with AC included hearing deficits, depression, pulmonary disorders, and abnormal serum sodium or potassium levels. Behavior patterns of acutely confused residents included hyperactive (n = 9, 31%), hypoactive (n = 8, 28%), and mixed (n = 7, 24%). In the majority of the AC cases, the etiology was multIfactorial infections and dehydration were the most common causes. Residents with AC had very poor 3-month outcomes. Thirty-four percent (n = 10) of the residents with AC died within 3 months of the evaluation. This study highlights the complexity and serious nature of AC in this frail population. PMID- 12741668 TI - The sleep experience of medical and surgical patients. AB - This study described and compared the sleep experience of medical and surgical patients during a hospital stay. During 3 consecutive nights, patients (n = 110) self-reported sleep quality using the Verran and Snyder Sleep Scale (VSH) and potentially disruptive factors using items from the Factors Influencing Sleep Questionnaire (FISQ). Surgical patients, on the first night, received more procedural care (p = .001), less sedative medication (p < .001), reported more sleep disturbance (p = .02), less sleep effectiveness (p = .03), and more need for sleep supplementation (p = .03). Variance in sleep effectiveness was explained by the FISQ score, age, and length of time in hospital (F = 6.86, p < .001). The sleep experience of patients varies between diagnostic groupings and across the hospital stay. Unit environmental and personal factors, factors that are amenable to therapeutic interventions, strongly influence the sleep experience. PMID- 12741669 TI - What's new on defining diarrhea in tube-feeding studies? AB - Nurses who are involved in studies of tube feeding tolerance or who review the literature on this topic are confronted with a myriad of definitions and methods of reporting diarrhea. In a 1992 study, the authors reported that these definition differences influenced results. In a review of the current literature, they determined that little progress had been made toward standardizing definitions and reports of diarrhea in studies of tube feeding. A secondary analysis of stool characteristics of hospitalized patients using various definitions of diarrhea showed there was a positive association between stool frequency and consistency. Criteria for stool consistency in the definitions of diarrhea appeared to have a greater influence on diarrhea outcomes when stool frequency was low. The authors suggest recognition of a taxonomy of definitions of diarrhea in the absence of consensus, which will help guide the design of future investigations and facilitate the evaluation and utilization of research. PMID- 12741670 TI - Pseudotaxonicity in MAMBAC and MAXCOV analyses of rating-scale data: turning continua into classes by manipulating observer's expectations. AB - Taxometric procedures such as mean above minus below a cut and maximum covariance can determine whether a trait is distributed as a discrete latent class. These methods have been used to infer taxonic structure in several personality and psychopathology constructs, often from analyses of rating scale data. This is problematic given (a) well established biases in ratings, (b) the human tendency to think categorically, and (c) implicit typological models of personality and psychopathology among expert raters. Using an experimental method in which the cognitive sets of raters were manipulated as dimensional versus categorical, it is demonstrated that pseudotaxonicity can be created readily with rating scale measures. This suggests that researchers avoid an exclusive reliance on rating scales when conducting taxometrics investigations. PMID- 12741671 TI - On specifying the null model for incremental fit indices in structural equation modeling. AB - In structural equation modeling, incremental fit indices are based on the comparison of the fit of a substantive model to that of a null model. The standard null model yields unconstrained estimates of the variance (and mean, if included) of each manifest variable. For many models, however, the standard null model is an improper comparison model. In these cases, incremental fit index values reported automatically by structural modeling software have no interpretation and should be disregarded. The authors explain how to formulate an acceptable, modified null model, predict changes in fit index values accompanying its use, provide examples illustrating effects on fit index values when using such a model, and discuss implications for theory and practice of structural equation modeling. PMID- 12741672 TI - Separating trait effects from trait-specific method effects in multitrait multimethod models: a multiple-indicator CT-C(M-1) model. AB - An overview of several models of confirmatory factor analysis for analyzing multitrait-multimethod (MTMM) data and a discussion of their advantages and limitations are provided. A new class of multi-indicator MTMM models combines several strengths and avoids a number of serious shortcomings inherent in previously developed MTMM models. The new models enable researchers to specify and to test trait-specific-method effects. The trait and method concepts composing these models are explained in detail and are contrasted with those of previously developed MTMM models for multiple indicators. The definitions of the models are explained step by step, and a practical empirical application of the models to the measurement of 3 traits x 3 methods is used to demonstrate their advantages and limitations. PMID- 12741673 TI - Information criteria for pairwise comparisons. AB - A new approach is presented for the interpretation of differences among means and proportions. Post hoc techniques, such as Tukey's honestly significant difference procedure, have interpretive problems related to intransitive decisions and technical issues arising from unequal sample sizes or heterogeneity of variance. These concerns can be avoided by considering ordered subsets of means and by using information criterion to select among competing models. This paired comparisons information-criterion (PCIC) approach is wholistic in nature and does not depend on interpreting a series of statistical tests. Simulation results suggest that a protected version of the PCIC procedure is desirable to minimize failures to detect the null case. This technique is illustrated for independent means, proportions, and means from repeated measures. PMID- 12741674 TI - Diagnosing item score patterns on a test using item response theory-based person fit statistics. AB - Person-fit statistics have been proposed to investigate the fit of an item score pattern to an item response theory (IRT) model. The author investigated how these statistics can be used to detect different types of misfit. Intelligence test data were analyzed using person-fit statistics in the context of the G. Rasch (1960) model and R. J. Mokken's (1971, 1997) IRT models. The effect of the choice of an IRT model to detect misfitting item score patterns and the usefulness of person-fit statisticsfor diagnosis of misfit are discussed. Results showed that different types of person-fit statistics can be used to detect different kinds of person misfit. Parametric person-fit statistics had more power than nonparametric person-fit statistics. PMID- 12741675 TI - A coefficient alpha for test-retest data. AB - Transient errors are caused by variations in feelings, moods, and mental states over time. If these errors are present, coefficient alpha is an inflated estimate of reliability. A true-score model is presented that incorporates transient errors for test-retest data, and a reliability estimate is derived. This estimate, referred to as the test-retest alpha, is less than coefficient alpha if transient error is present and is less susceptible to effects due to item recall than a test-retest correlation. An assumption underlying the test-retest alpha is essential tau equivalency of items. A test-retest split-half coefficient is presented as an alternative to the test-retest alpha when this assumption is violated. The test-retest alpha is the mean of all possible test-retest split half coefficients. PMID- 12741676 TI - Estimating the reliability of a test split into two parts of equal or unequal length. AB - When the reliability of test scores must be estimated by an internal consistency method, partition of the test into just 2 parts may be the only way to maintain content equivalence of the parts. If the parts are classically parallel, the Spearman-Brown formula may be validly used to estimate the reliability of total scores. If the parts differ in their standard deviations but are tau equivalent, Cronbach's alpha is appropriate. However, if the 2 parts are congeneric, that is, they are unequal in functional length or they comprise heterogeneous item types, a less well-known estimate, the Angoff-Feldt coefficient, is appropriate. Guidelines in terms of the ratio of standard deviations are proposed for choosing among Spearman-Brown, alpha, and Angoff-Feldt coefficients. PMID- 12741677 TI - Genetic alterations responsible for metastatic phenotypes of lung cancer cells. AB - It is now widely accepted that human carcinogenesis is a multi-step process and phenotypic changes during cancer progression reflect the sequential accumulation of genetic alterations in cells. Thus, in order to understand the process of acquisition of metastatic phenotypes in cancer cells, it is indispensable to identify genes whose alterations accumulate during cancer progression and correlate with metastatic phenotypes of cancer cells. For this reason, we have been searching for genes that are preferentially altered in metastatic lung cancer cells and have activities to regulate their metastatic potentials. In lung cancer, both the p16INK4A/RB and p53 genes are frequently inactivated and are critical determinants for the regulation of cell growth and apoptosis. However, it still remains unclear whether these genes are also involved in the regulation of metastatic potential in lung cancer cells. Recently, we identified a novel myosin family gene, MYO18B, from the chromosome 22q12.1 region which shows frequent loss of heterozygosity in advanced lung cancer, and we found that this gene is inactivated in approximately 50% of lung cancers by deletions, mutations and methylation. Furthermore, restoration of MYO18B expression suppressed anchorage-independent growth of lung cancer cells. Thus, it was indicated that the MYO18B gene is a strong candidate for a metastasis suppressor gene of human lung cancer. Further functional and biological studies of the MYO18B gene will help us understand the molecular pathway of human lung cancer progression. PMID- 12741678 TI - CD44 and its partners in metastasis. AB - The establishment of metastasis requires that tumor cells acquire new adhesion and migration properties to emigrate from primary sites and colonize distant organs. CD44 is a cell membrane protein often overexpressed on tumor cells and, being both a cell-cell and cell-extracellular matrix adhesion protein, is well positioned to contribute to this process. Furthermore the interaction of CD44 with other cellular proteins involved in motogenesis and proteolysis is a determinant factor in cell migration and invasion. This review summarizes current knowledge on the role of CD44 in metastasis, as well as the challenges on understanding how this process operates. PMID- 12741679 TI - Integrin adhesion receptors in tumor metastasis. AB - Integrins are a family of cell adhesion receptors that support and modulate a variety of cellular functions that are required for tumor metastasis. Integrins expressed by tumor cells and host cells can directly contribute to the control and progress of metastatic dissemination. During tumor development, changes in integrin expression, intracellular control of integrin functions and signals perceived from integrin ligand binding impact upon the ability of tumor cells to interact with their environment and enable metastatic cells to convert from a sessile, stationary to a migratory and invasive phenotype. Integrins are involved in each step of the metastatic cascade and affect tumor cell survival and interaction with changing environments in transit from the primary tumor to distant target organs. PMID- 12741680 TI - Angiogenesis and the role of epigenetics in metastasis. AB - The major obstacle to devising effective ways to treat cancer is its heterogeneity and genetic instability. It was originally postulated that targeting the process of tumor angiogenesis could circumvent this problem, as it involves genetically stable epigenetically controlled host stroma. Thus, anti angiogenic approaches should be applicable across various tumor types and organ sites, including metastases. However, early clinical experience with this therapy revealed unexpectedly distinct responses between different tumors and organ sites. Here we propose that the heterogeneity of pre-clinical and clinical results obtained with anti-angiogenic agents stems from the deep functional linkage that may exist between genetic and epigenetic tumor progression. Thus, epigenetic processes regulating tumor associated host blood vessels (such as tumor microenvironment) display unstable, heterogeneous and progressive characteristics to an extent comparable with (and causally linked to) the instability of the cancer cell genome. As well, many known epigenetic factors (such as hypoxia, inflammation, expression of growth factors, etc.) may have genetic causes and consequences (e.g., oncogene expression, loss of tumor suppressor genes). This reciprocal interrelationship and heterogeneity may translate into site and stage specific changes in angiogenesis regulation, and angiogenesis dependence, ultimately to changes in the metastatic ability/efficiency of cancer cells, even in the same patient. A better understanding of the linkage between genetic and epigenetic events in growth and metastasis of various cancers may result in more effective use of anti-angiogenic therapy in future. PMID- 12741681 TI - The vascular phenotype of melanoma metastasis. AB - Melanoma cells but not normal melanocytes share important cell surface molecules with endothelial cells. From this observation has grown our hypothesis that melanoma and endothelial cells use the same cell surface molecules for invasion but that their intracellular signaling is different allowing the malignant cells dominance over the normal cells. Stromal fibroblasts and endothelial cells communicate with metastatic cells through an interactive network of cell-cell and cell-matrix signaling. Fibroblasts induce capillary-like differentiation of microvascular endothelial cell monolayers through cell-cell contact and soluble factors. It is then hypothesized that normal cellular precursors are recruited from the bone marrow to the site of a growing melanoma metastasis. Thus pathways for angiogenesis and vasculogenesis are guided through the cooperation of fibroblasts and melanoma cells perpetuated by the dominance of the metastatic melanoma cells. PMID- 12741682 TI - The hypoxic tumour microenvironment and metastatic progression. AB - The microenvironment of solid tumours contains regions of poor oxygenation and high acidity. Growing evidence from clinical and experimental studies points to a fundamental role for hypoxia in metastatic progression. Prolonged hypoxia increases genomic instability, genomic heterogeneity, and may act as a selective pressure for tumour cell variants. Hypoxia can also act in an epigenetic fashion, altering the expression of genes. Hypoxia-induced changes in gene expression alter non-specific stress responses, anaerobic metabolism, angiogenesis, tissue remodeling, and cell-cell contacts. Experimental studies have demonstrated that inhibition of proteins involved in these processes can modify metastasis formation, suggesting a causal role in metastatic progression. Recent advances in high-throughput screening techniques have allowed identification of many hypoxia induced genes that are involved in the processes associated with metastasis. Here we review the epigenetic control of gene expression by the hypoxic microenvironment and its potential contribution to metastatic progression. PMID- 12741683 TI - Transcriptional regulation of metastasis-related genes in human melanoma. AB - Melanoma begins with benign nevi and progresses to radial growth phase (RGP) and to vertical growth phase [(VGP), metastatic phenotype]. The molecular changes associated with these transitions are not yet well defined. However, transcriptional regulation of some genes that are critical in melanoma progression is beginning to be elucidated. The first part of this review will focus on our recent studies demonstrating that progression of human melanoma is associated with loss of expression of the transcription factor AP-2. In metastatic melanoma cells, this loss resulted in overexpression of MCAM/MUC18 and MMP-2, and lack of expression of c-KIT. In further investigations, we inactivated AP-2 in SB-2 primary cutaneous melanoma cells by using a dominant-negative AP-2, the AP-2B gene. Expression of AP-2B in SB-2 cells augmented their tumorigenicity in nude mice and upregulated MMP-2 expression and activity. We have also recently demonstrated that loss of AP-2 expression in metastatic melanoma cells resulted in overproduction of the thrombin receptor, PAR-1. Other studies have shown that AP-2 regulates additional genes involved in melanoma development and progression, including E-cadherin, p21/WAF-1, HER2, Bcl-2, FAS/APO-1, IGF-R-1, and VEGF. We propose that loss of AP-2 is crucial in the development of malignant melanoma. Additionally, the transition of melanoma cells from RGP to VGP is associated with overexpression of two transcription factors, CREB and ATF-1, both of which may act as survival factors for human melanoma cells. The second part of the review will briefly discuss the role of other transcription factors, including ATF-2, SNAIL, MITF, and NFkappaB in the progression of human melanoma and will summarize recent knowledge on how changes in the expression of these transcription factors contribute to acquisition of the metastatic phenotype in human melanoma. PMID- 12741684 TI - Translational control and metastatic progression: enhanced activity of the mRNA cap-binding protein eIF-4E selectively enhances translation of metastasis-related mRNAs. AB - To form metastases, tumors must break from the primary tumor site, invade surrounding tissues, enter and survive within the circulation and ultimately colonize a distal tissue. Each of these steps requires the cooperative function of numerous proteins--proteins that facilitate angiogenesis (e.g., VEGF), cell survival (e.g., Bcl-2), invasion (e.g., MMPs), and autocrine growth stimulation (e.g., c-myc, cyclin D1). Although expression of these proteins is regulated at many levels by disparate stimuli, translation of these key malignancy-related proteins is regulated primarily by the activity of the mRNA cap-binding protein eIF-4E, the rate-limiting member of the eIF-4F translation initiation complex. By binding the cap structure at the 5' terminus of cellular mRNAs, eIF-4E recruits mRNAs to the eIF-4F complex, which then scans from the 5' cap through the untranslated region (5'UTR), unwinding secondary structure to reveal the translation initiation codon and to enable ribosome loading. Messenger RNAs with short unstructured 5' UTRs are more easily translated than mRNAs harboring lengthy, highly structured 5' UTRs, as these prohibit efficient scanning and start codon recognition. As such, the translation of these mRNAs, which typically encode proteins involved in angiogenesis (e.g., VEGF), tumor growth (cyclin D1) and survival (Bcl-2), is suppressed except when eIF-4E is engaged with the eIF-4F complex--a common event in many human and experimental cancers. This review focuses on the hypothesis that enhanced eIF-4E function contributes to metastatic progression by selectively upregulating the translation of key malignancy-related proteins that together conspire to drive the metastatic process. PMID- 12741685 TI - The skeleton as a unique environment for breast cancer cells. AB - Bone is a favored location for several cancer metastases especially breast, prostate and myeloma. This review evaluates various properties of the skeleton that contribute to its successful colonization by breast cancer cells. The first consideration is the unique aspects of the vasculature of metaphyseal bone, which may account for the initial lodging of breast cancer cells in specific regions of the skeleton. Metasphyseal bone, found at the ends of long bone, in ribs and in vertebrae, is comprised of trabecular bone interspersed with marrow and a rich vasculature. The chemotactic factors that arise from bone marrow and bone cells are discussed in terms of cancer cell migration out of the vasculature and entry of cancer cells into the marrow cavity. Once the breast cancer cells have migrated into the metaphysis, they interact both directly and indirectly with bone cells and other cells in the marrow. As tumor growth progresses, functional bone cells are lost, most likely through apoptosis. PMID- 12741686 TI - Social contexts: transcending their power and their fragility. AB - For more than four decades, my colleagues and I have examined how social contexts and individuals influence each other. To provide a perspective on this work, I describe some lessons we have learned: situational influence exemplifies both the power and fragility of social contexts; common aspects of settings underlie their power, for better and for worse; individuals' health and well-being is affected by powerful forces in eight domains; and intervention programs are powerful settings that encompass risks as well as rewards. I then note some unresolved questions, such as how to balance the risks and rewards of powerful environments, how to better understand the interplay between individuals and the social contexts they select and create, how to learn from the fact that many people overcome the power of traumatic social contexts, and how to apply our knowledge to balance the risks and rewards of individual and community contexts. PMID- 12741687 TI - Correlates of African American and Latino parents' messages to children about ethnicity and race: a comparative study of racial socialization. AB - Recently, social scientists have become increasingly interested in the nature of communications from parents to children regarding ethnicity and race. Termed racial socialization, race-related messages to children may have important consequences for children's identity development and well-being. This study examined the frequency and correlates of two dimensions of racial socialization messages about ethnic pride, history, and heritage (Cultural Socialization) and messages about discrimination and racial bias (Preparation for Bias)--among 273 urban African American, Puerto Rican, and Dominican parents. Parents reported more frequent Cultural Socialization than Preparation for Bias. There were no significant ethnic group differences in the frequency of Cultural Socialization. However, African American parents reported more frequent Preparation for Bias than did Dominican parents who, in turn, reported more frequent messages of this sort than did Puerto Rican parents Ethnic identity was a stronger predictor of Cultural Socialization among Puerto Rican and Dominican parents than among their African American counterparts. In contrast, perceived discrimination experiences was a stronger predictor of Preparation for Bias among African American and Dominican parents than among Puerto Rican parents. Finally, race-related phenomenon accounted for more variance in both Cultural Socialization and Preparation for Bias among parents reporting on their behaviors with children 10 17 years old as compared to parents reporting on their behaviors with children 6 9 years old. PMID- 12741688 TI - Neighborhood structure, parenting processes, and the development of youths' externalizing behaviors: a multilevel analysis. AB - Associations among neighborhood structure, parenting processes, and the development of externalizing behavior problems were investigated in a longitudinal sample of early adolescents (from age 11 to 13). Mothers' reports of parental monitoring (at age 11), mothers' and youths' reports of the amount of youths' unsupervised time (at age 11), and youths' reports of positive parental involvement (at age 12) were used to predict initial levels (at age 11) and growth rates in youths' externalizing behavior as reported by teachers. Census based measures of neighborhood structural disadvantage, residential instability, and concentrated affluence were expected to moderate the effects of parenting processes (e.g., parental monitoring) on externalizing behavior. Hierarchical linear modeling results revealed that less parental monitoring was associated with more externalizing behavior problems at age 11, and more unsupervised time spent out in the community (vs. unsupervised time in any context) and less positive parental involvement were associated with increases in externalizing behavior across time. Furthermore, the decrease in externalizing levels associated with more parental monitoring was significantly more pronounced when youths lived in neighborhoods with more residential instability. PMID- 12741690 TI - The quest for a liberating community psychology: an overview. PMID- 12741689 TI - Prevention science and neighborhood influences on low-income children's development: theoretical and methodological issues. AB - Despite the rapid growth of research on neighborhood influences on children, little of this research may be useful to prevention scientists. Most studies have ignored processes by which neighborhood conditions influence individual outcomes. To encourage neighborhood research that can better guide the development of preventive interventions, we propose a model that focuses attention on mediating and moderating processes, is appropriate for studies interested in individual differences in outcomes, acknowledges the transactions between residents and neighborhoods, and is sensitive to how neighborhood influences may differ for children at different developmental stages. Furthermore, we argue that greater attention to several methodological issues also can make neighborhood research more useful for the next generation of prevention programs to help low-income urban families and children cope successfully with the challenges posed by their neighborhoods. PMID- 12741691 TI - Narrating survival and change in Guatemala and South Africa: the politics of representation and a liberatory community psychology. AB - Peace accords and international interventions have contributed to the suspension of armed conflict and the censuring of repressive regimes in many parts of the world. Some governments and their opposition parties have agreed to the establishment of commissions or other bodies designed to create historical records of the violations of human rights and foster conditions that facilitate reparatory and reconciliatory processes. This paper explores selected roles that community psychologists have played in this process of remembering the past and constructing new identities towards creating a more just future. With reference to two community groups (in Guatemala and South Africa) we show how efforts to "speak out" about one's own experiences of political and military repression involve complex representational politics that go beyond the simple binary opposition of silencing versus giving voice. The Guatemalan group consisted of Mayan Ixil women who, together with the first author, used participatory action research and the PhotoVoice technique to produce a book about their past and present struggles. The South African group, working within the ambit of the Truth and Reconciliation Commission and in collaboration with the third author and others, explored ways of speaking about their roles in apartheid and post apartheid society. Although both these initiatives can be seen as moments in on going struggles to overcome externally-imposed repressive practices that censor the voices of marginalized communities, they also serve to dispel overly romanticized notions of "univocal" communities now liberated to express themselves in an unmediated and unequivocal fashion. The paper discusses how each group of women instead entered into subtly nuanced relationships with community psychologists involving a continual interplay between the authenticity of their self-representational accounts and the requirements of the discursive technologies into which they were being inducted and the material conditions within their sites of struggle. In both cases the group's agenda also evolved over time, so that what emerged was not so much a particular account of themselves, or even the development of a particular "voice" for speaking about themselves, but an unfolding process--for the groups and for the community psychologists who accompanied them--of becoming active players in the postmodern, mediated world of self-representational politics and social struggle. PMID- 12741692 TI - Bridging the personal and the political: practices for a liberation psychology. AB - In the Irish context, legacies of colonialism, the Northern Ireland conflict situation, and the strength of community and women's liberation movements all provide rich resources for understanding the processes involved in both oppression and liberation. This paper draws on the theoretical and research literature and on Irish experiences to develop an understanding of some of the processes and practices that aid in liberation. The research is grounded in diverse writings on oppression and liberation, which include writings on colonialism (E. Duran & B. Duran, 1995; F. Fanon, 1967; V. Kenny, 1985, L. Maracle, 1996), feminist psychology (J. B. Miller, 1986; S. Wilkinson, 1996), liberation psychology (H. A. Bulhan, 1985; L. Comas-Diaz, M. B. Lykes, & R. D. Alarcon, 1998; I. Martin-Baro 1994; Starhawk, 1987), and psychological aspects of racism (b. hooks, 1993; A. Mama, 1995; R. J. Watts, D. M. Griffith, & J. Abdul Adil, 1999), homophobia (A. R. D'Augelli & C. J. Patterson, 1995), poverty (K. O'Neill, 1992), and other dimensions of oppression. PMID- 12741693 TI - The challenge of a positive self-image in a colonial context: a psychology of liberation for the Puerto Rican experience. AB - Community psychology and the psychology of liberation provide a framework to analyze colonization. Puerto Rico has always been a colony. This experience has direct negative effects on Puerto Ricans' national identities and their emotions. The purpose of this study was to explore emotions associated with Puerto Rican national identities in a sample of 35 high school students. While participating in 4 focus groups, the youths discussed negative and positive emotions associated to their national identities. The most frequent ones were shame and pride. They also identified situations which promoted negative and positive emotions. The role of a community psychology of liberation in understanding these phenomena as well as in embracing resistance to colonization is discussed. PMID- 12741694 TI - Identity and oppression: differential responses to an in-between status. AB - Oppression operates at various levels, with varying degrees of negativity, and groups respond in markedly different ways. In this paper, the in-between status of the colored South African group is used to illustrate issues of identity and oppression under the Apartheid system-and differing ways in which oppression was experienced and used. The colored group had many social advantages over Blacks, but were also used to oppress that group. Habituation, accommodation, and relative advantage were identified as dynamics within the broader context of power and privilege that contributed to cultural and psychological marginality and status ambivalence of the coloreds. These processes must be understood within the historical, social, and political context of the community. What is evident from the data is that groups and individuals can take up various positions along a continuum of oppressor-oppressed, depending upon the contexts, time, and social and legal relationships involved in their interactions. PMID- 12741695 TI - The violent matrix: a study of structural, interpersonal, and intrapersonal violence among a sample of poor women. AB - One goal of this paper is to present an integrated tripartite model of violence, with a focus on structural violence within an oppression paradigm. Using qualitative and quantitative data from 27 women (70% African American and 30% European American) who participated in a national substance abuse treatment demonstration program, we describe a model of violence in which structural violence is presented within a transactional relationship with interpersonal, and intrapersonal violence. We suggest that the effects of structural, interpersonal, and intrapersonal violence are magnified when race and poverty are considered. The second goal of the paper is to present a preliminary test of the new model of violence. Results indicated that different levels and types of violence are interrelated. Implications of these findings for empowering solutions are suggested. PMID- 12741696 TI - Integrating psychological research on girls with feminist activism: a model for building a liberation psychology in the United States. AB - A liberation psychology is needed to bridge the gap between psychology's focus on individual distress and broad social forces that foster such distress. We offer a model for bridging this gap by focusing on a specific area of psychology (psychological research on girls) and a specific social movement (feminist activism). Psychological research on girls and feminist activism share the common goal of improving the lives of girls and women. However, both have fallen short of this goal. This is due, in part, to the weaknesses associated with each endeavor and to the fact that the complementary strengths of each have remained isolated from the other. In this paper, we propose a common language and shared framework to integrate psychological research with feminist activism. First, we review the basic strengths and weaknesses associated with psychological research and feminist activism, with a particular focus on how they are distinct from one another. Second, we provide a taxonomic framework for integrating these two areas on the basis of the stress paradigm, with specific examples provided from our recent reviews of the literature and our own empirical work with adolescent girls. Finally, we conclude with recommendations for future work needed to integrate psychological research on girls with feminist activism toward the goal of building a liberation psychology in the United States. PMID- 12741697 TI - Emancipatory responses to oppression: the template of land-use planning and the Old Order Amish of Ontario. AB - In this paper I discuss the clash of values between the Old Order Amish community of Ontario and the dominant social paradigm in agriculture. Land-use and agricultural regulations, designed for an industrial style of agriculture, are experienced as a threat to the survival of the Old Order Amish agricultural social economy and community. The paper describes how I have worked with the Old Order Amish to respond to these challenges and to create public policies that will allow them to maintain their human and agricultural diversity and small scale sustainable farm practices. Four case studies illuminating the oppressive land-use regulations along with the emancipatory responses to the oppression are examined. The social transformation themes include principles and processes for community psychologists, land-use planners, and community economic development practitioners to consider. The article has heuristic value for a practice-based approach to social change. PMID- 12741698 TI - Emancipatory education versus school-based prevention in African American communities. AB - Schools have become strategic settings for the work of community psychologists. In a review of 177 primary prevention programs for children and adolescents, Durlak and Wells (1997) found that 129 (72.9%) were based in schools. The literature in community psychology describes many school-based prevention programs targeting problems such as substance abuse, school "maladjustment," delinquency, and violence (e.g. C. A. Mason, A. M. Cauce, L. Robinson & G. W. Harper, 1999). A large number of these programs are based in schools in African American communities and include social-cognitive, decision making, affective education, and other skills-building modules along with direct instruction. In this paper, it is argued that ideas from emancipatory education (e.g. Freire, 1998) and African-centered education (e.g. H. Madhubuti & S. Madhubuti, 1994; M. J. Shujaa, 1995) should guide school-based interventions in communities of people of African descent. There is an extensive and distinguished history of emancipatory schools and school-based programs in African American communities. Included in this history are the freedom schools during reconstruction, the SNCC Freedom Schools, the Liberation Schools of the Black Panther Party, the Malcolm X Academy in Detroit, Sankofa Shule in Lansing, the Institute for Positive Education/New Concept Development Center in Chicago, the Benjamin E. Mays Institute in Hartford, and the schools affiliated with the Council of Independent Black Institutions (CIBI) to name just a few. This paper will first provide a brief, critical review of the role of schools and social oppression. Second, primary prevention programs in communities of people of African descent will be examined, questioning some of the dominant methods and assumptions. Next, underlying assumptions about relationships between African identity, educational success, and healthy outcomes for young people will be addressed. This will be followed by a discussion of African-centered emancipatory education, focusing specifically on the role of students as agents of social change and the importance of critical reflection on African cultural resources. The Benjamin E. Mays Institute will be presented as an example of how ideas from an African centered emancipatory approach to education have been incorporated within a school serving a community of people of African descent in Hartford, Connecticut. PMID- 12741699 TI - Sociopolitical development. AB - This paper examines theories and concepts relevant to sociopolitical development (SPD). As an emerging theory, SPD expands on empowerment and similar ideas related to social change and activism in community psychology--oppression, liberation, critical consciousness, and culture among them. SPD is the process by which individuals acquire the knowledge, analytical skills, emotional faculties, and the capacity for action in political and social systems necessary to interpret and resist oppression. Equally as important is a vision of liberation that is an alternative to oppressive conditions. All of these concepts have been underemphasized in the social change literature of U.S. community psychology. In our view, sociopolitical development is vital to human development and the creation of a just society. As part of identifying and illustrating concepts and processes relevant to SPD theory, we will draw from the words of young African American activists who were interviewed as part of a research study. PMID- 12741700 TI - Understanding, resisting, and overcoming oppression: toward psychopolitical validity. AB - My first objective in this paper is to synthesize, synoptically, the literature on oppression and liberation with the contributions to this special issue. To fulfil this aim I introduce a framework for understanding, resisting, and overcoming oppression. The framework consists of psychopolitical well-being; experiences, consequences, and sources of oppression; and actions toward liberation. Each of these components is subdivided into 3 domains of oppression and well-being: collective, relational, and personal. Experiences of suffering as well as resistance and agency are part of the framework. My second objective is to offer ways of closing the gap between research and action on oppression and liberation. To do so I suggest 2 types of psychopolitical validity: epistemic and transformative. PMID- 12741701 TI - Implantable cardioverter defibrillator in high-risk long QT syndrome patients. AB - INTRODUCTION: Implantable cardioverter defibrillators (ICDs) are increasingly being used in high-risk long QT syndrome (LQTS) patients, but there are limited data regarding clinical experience with this therapeutic modality. The aim of this study is to describe the clinical characteristics of 125 LQTS patients treated with ICDs compared with LQTS patients having similar risk indications who were not treated with ICDs. Among 125 LQTS patients with ICDs, there were 54 cardiac arrest survivors, 19 patients who had ICDs implanted due to recurrent syncope despite beta-blocker therapy, and 52 patients with ICDs implanted due to other reasons, including syncope and LQTS-related sudden death in a close family member. Patients with cardiac arrest and those with recurrent syncope despite beta-blocker therapy (n = 73) were compared to 161 LQTS patients who had similar indications (89 cardiac arrest and 72 recurrent syncope despite beta-blocker therapy) but did not receive ICDs. Total mortality was the endpoint of the analysis. There was 1 (1.3%) death in 73 ICD patients followed an average of 3 years, whereas there were 26 deaths (16%) in non-ICD patients during mean 8-year follow-up (P = 0.07 from log rank test from Kaplan-Meier curves). CONCLUSION: ICDs provide an important therapeutic option to prevent sudden arrhythmic death in high-risk LQTS patients. A long-term prospective study is needed to determine the benefit of this therapeutic modality in LQTS patients. PMID- 12741702 TI - Saving lives in congenital long QT syndrome: who benefits from implantable cardioverter defibrillator therapy? PMID- 12741703 TI - Implantable cardioverter defibrillator lead complications and laser extraction in children and young adults with congenital heart disease: implications for implantation and management. AB - INTRODUCTION: Implantable cardioverter defibrillators (ICDs) are being implanted for primary and secondary prevention of sudden death in children and young adults with congenital heart disease. Over time, ICD leads adhere to venous endothelium and endocardium. Lead removal, when necessary, often requires disruption of this fibrous tissue. METHODS AND RESULTS: We retrospectively reviewed and analyzed our experience with ICD lead extraction in children and young adults with congenital heart disease. From April 1999 through January 2002, 14 patients underwent 15 lead extraction procedures to remove 21 leads (17 ICD leads and 4 pacing or sensing leads). Seven patients had surgically corrected structural heart disease (5 transposition of the great arteries with atrial switch repair and 2 corrected tetralogy of Fallot). Mean patient age at extraction was 17.9 +/- 5.7 years (range 9-32), and mean duration of lead implantation was 42.0 +/- 18.9 months (range 15-75). Fourteen of 15 procedures were performed for lead fracture or failure. A laser sheath was used for 20 of 21 lead extractions. Twenty of 21 leads (95%) were completely extracted. There were three instances of blood loss requiring transfusion. There were no major complications or deaths. CONCLUSION: Young congenital heart disease patients with an ICD are at risk for growth related lead distortion. The use of a laser sheath is safe and effective for ICD lead extraction in congenital heart disease patients, despite coil adherence and altered anatomy. It may be advisable to avoid dual-coil leads in patients with the potential for future growth. PMID- 12741704 TI - P wave polarities of an arrhythmogenic focus in patients with paroxysmal atrial fibrillation originating from superior vena cava or right superior pulmonary vein. AB - INTRODUCTION: The superior vena cava (SVC) and right superior pulmonary vein (RSPV) are anatomically close structures. Using 12-lead ECG may facilitate identification of ectopic foci from SVC or RSPV. The aim of this study was to assess whether P wave polarity on surface ECG is helpful in distinguishing an arrhythmogenic focus of paroxysmal atrial fibrillation (AF) from SVC or RSPV. METHODS AND RESULTS: Thirty-four patients with paroxysmal AF from the SVC (group I: 17 patients, 10 men and 7 women; mean age 57 +/- 12 years) or RSPV (group II: 17 patients, 15 men and 2 women, mean age 62 +/- 14 years) underwent electrophysiologic study and radiofrequency (RF) catheter ablation. All of the AF foci were confirmed by successful ablation. P wave polarities on surface ECG inferior leads were positive during sinus rhythm and ectopic beats in both groups. Leads I, aVR, aVL, and V1 were further analyzed. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) in predicting an arrhythmogenic focus of AF from SVC or RSPV were provided. P wave polarity in lead aVR was negative in all 34 patients. P wave polarity in lead V1 was positive in 47.1% of SVC ectopy but positive in all RSPV ectopy. The combination of a biphasic or isoelectric P wave polarity in lead V1 or a biphasic P wave polarity in lead aVL had a sensitivity of 71%, specificity of 82%, PPV of 80%, and NPV of 74% in predicting an arrhythmogenic focus of AF from SVC. CONCLUSION: P wave polarity in leads V1 and aVL may predict an arrhythmogenic focus of AF from SVC or RSPV. PMID- 12741705 TI - Efficacy and safety of targeted focal ablation versus PV isolation assisted by magnetic electroanatomic mapping. AB - INTRODUCTION: Pulmonary vein (PV) triggers initiate atrial fibrillation (AF). The aim of this study was to compare the outcome of focal PV ablation versus targeted PV electrical isolation aided by multipolar catheter recordings in the coronary sinus (CS) and right atrium and magnetic electroanatomic mapping (MEAM) for drug refractory AF. METHODS AND RESULTS: Multipolar recordings identified PVs with triggers based on PV ostial pace map match for spontaneous and provoked triggers. PV triggers were provoked by isoproterenol, adenosine, and AF induction followed by cardioversion. MEAM defined PV ostial anatomy and assisted in localization of AF trigger and ablation lesions. All focal PV ablation procedures preceded PV isolation procedures at our institution. To limit a learning curve effect and validate the comparison, the results included outcome of procedures by a single experienced operator in the last 32 consecutive patients undergoing focal PV ablation and in 75 consecutive patients undergoing PV isolation. Patient characteristics were similar with respect to mean age (50 vs 52 years), mean left atrial size (4.3 vs 4.2 cm), presence of paroxysmal AF (84% vs 88%), and demonstration of non-PV triggers (16% in both groups). PV isolation was confirmed in 99% of PVs by multipolar circular catheter. MEAM confirmed noncircumferential ostial ablation in 69% of PVs. Patients undergoing PV isolation had less AF from PV triggers at the end of ablation (1% vs 16%, P < 0.01); had less AF at 2 months (17% vs 42%, P < 0.001); and had 1-year freedom from AF of 80% versus 45% (P < 0.001). Adverse events were low in both groups with no stroke or symptomatic PV stenosis. CONCLUSION: Using the described techniques, PV electrical isolation of PVs demonstrating spontaneous and/or provoked triggers is superior to focal PV ablation, with marked differences in outcome by 2 months. MEAM confirmed the noncircumferential nature of ostial ablation for effective isolation of most PVs and may play a role in the low risk and good outcome observed. The good outcome of targeted PV isolation as described suggests the need for a prospective comparison of targeted versus empiric PV isolation techniques. PMID- 12741706 TI - Total pulmonary vein occlusion as a consequence of catheter ablation for atrial fibrillation mimicking primary lung disease. AB - INTRODUCTION: Catheter ablation has recently been used for curative treatment of atrial fibrillation. METHODS AND RESULTS: Three of 239 patients who underwent ablation close to the pulmonary vein (PV) ostia at our institute developed severe hemoptysis, dyspnea, and pneumonia as early as 1 week and as late as 6 months after the ablation. Because the patients were arrhythmia-free, the treating physician initially attributed the symptoms to new-onset pulmonary disease (e.g., bronchopulmonary neoplasm). After absent PV flow was confirmed by transesophageal echocardiography, transseptal contrast injection depicted a totally occluded PV in all three patients. Successful recanalization, even in chronically occluded Pvs, was performed in all patients. During follow-up, Doppler flow measurements by transesophageal echocardiography demonstrated restenosis in all primarily dilated PV, which led to stent implantation. CONCLUSION: PV stenosis/occlusion after catheter ablation of atrial fibrillation occurs in a subset of patients. However, because in-stent restenosis occurred in two patients after 6 to 10 weeks, final interventional strategy for PV stenosis or occlusion remains unclear. To prevent future PV stenosis or occlusion, a decrease in target temperature and energy of radiofrequency current or the use of new energy sources (ultrasound, cryothermia, microwave) seems necessary. PMID- 12741707 TI - Pulmonary vein occlusion/stenosis after pulmonary vein ablation for atrial fibrillation. PMID- 12741708 TI - Site-specific arrhythmogenesis in patients with Brugada syndrome. AB - INTRODUCTION: It has been believed that electrophysiologic abnormality of the epicardial region of the right ventricular free wall may play an important role in arrhythmogenesis of phase 2 reentry in Brugada syndrome, but clinical evidence of the occurrence of ventricular arrhythmias at the right ventricular free wall has not been evaluated. In this study, we evaluated the site-specific inducibility of ventricular fibrillation (VF) and the origin of spontaneous premature ventricular contractions (PVCs) in patients with Brugada syndrome. METHODS AND RESULTS: Forty-five patients with Brugada-type ECG were enrolled in this study. Spontaneous PVCs were recorded in 9 patients. Programmed electrical stimulation (PES) was performed at the right ventricular apex (RVA), the free wall and septal region of the right ventricular outflow tract (RVOT), and the left ventricle (LV). The inducibility of PVT/VF was evaluated at each ventricular site, and the origin of PVC was determined by pace mapping. Sustained VF was induced in 17 patients. VF was induced in all 17 patients by PES at RVOT. Although PES at the septal region of the RVOT induced VF in only 5 patients (29%), PES at the free-wall region of the RVOT induced PVT/VF in 13 patients (76%). PES at RVA induced VF in only 2 patients (12%), and PES at LV failed to induce any arrhythmic events. Ventricular pace mapping showed that 64% of PVCs occurred at the free-wall region of the RVOT, 18% at the septal region of the RVOT, 9% at RVA, and 9% at LV. CONCLUSION: VF in patients with Brugada syndrome frequently is induced at the free-wall region of the RVOT area. The origin of PVC appears to be related to the site of PVT/VF induction by PES. PMID- 12741709 TI - Site-specific arrhythmogenesis in Brugada syndrome? PMID- 12741711 TI - Genetic circuits and electric circuits: development of the cardiac conduction system. PMID- 12741710 TI - Development of the cardiac conduction system as delineated by minK-lacZ. AB - INTRODUCTION: Due to the lack of good molecular markers, for decades the morphogenetic origin of the cardiac conduction system has been a matter of debate. More recently, the spatial expression of minK-lacZ in the adult mouse heart has been shown, for the larger part, to be coincident with the conduction tissues. METHODS AND RESULTS: To trace the embryonic development of this system, we performed an analysis of the expression of this construct throughout early cardiac development. Expression was first seen at the eighth embryonic day. Subsequently, discrete rings were found at the sinuatrial, atrioventricular, interventricular, and ventriculoarterial junctions. With time, the expression became restricted to boundary regions of the heart, such as the hinges of the leaflets of the pulmonary and aortic valves, the atrioventricular rings, and the venous valves, as well as becoming incorporated into the definitive conduction tissues themselves. In the postnatal heart, the areas retaining minK-lacZ positivity outside of the definitive conduction tissues are known to be the site of origin of abnormal cardiac rhythms, suggesting that ectopic foci may derive from tissues that share a common developmental pathway with the definitive conduction system. CONCLUSION: Our findings suggest that the boundary regions between compartments, along with the atrioventricular conduction axis, share a common developmental pathway. PMID- 12741712 TI - Transmural action potential changes underlying ventricular electrical remodeling. AB - INTRODUCTION: Although it is well established that alterations in heart rate or activation sequence induce electrical remodeling in the atria, electrical remodeling in the ventricle is poorly understood. METHODS AND RESULTS: To determine the changes in cellular repolarization that underlie ventricular electrical remodeling caused separately by altered heart rate and activation sequence, optical action potentials were recorded simultaneously from 256 sites spanning the transmural wall of the arterially perfused canine wedge preparation (n = 15). Action potentials were compared from the same sites under identical conditions [endocardial pacing, cycle length (CL) = 1,000 msec], before and after an intervening 20- to 60-minute period of remodeling induced by (1) rapid pacing (CL = 300 msec) with no change in activation sequence; (2) altered activation sequence (epicardial pacing) with no change in rate; or (3) no change in rate or activation sequence (control). Action potential duration (APD) shortened by 24.8 +/- 4.8 msec following a period of rapid heart rate (P < 0.05) but prolonged (by 12.7 +/- 1.8 msec) following a period of altered activation sequence (P < 0.05). Hence, even after restoration of baseline heart rate and activation sequence, there were persistent changes in APD from baseline, indicative of electrical remodeling. Moreover, the orientation of the maximum APD gradient across the transmural wall changed more significantly following heart rate remodeling (by 27.7 degrees +/- 4.9 degrees, P < 0.05) than following activation sequence remodeling (by 12.3 degrees +/- 2.4 degrees, P < 0.05). CONCLUSION: Persistent changes in ventricular repolarization can be induced by surprisingly short periods of altered rate or activation sequence. In contrast to atrial remodeling, electrical remodeling in the ventricle can result in prolonged APD (with altered activation sequence) or reversal of APD gradient orientation (with rapid rate), suggesting that the nature of ventricular electrical remodeling induced by these two perturbations is different. PMID- 12741713 TI - Accessory pathway in left inferoposterior diverticulum masquerading as left posterior pathway due to conduction over coronary sinus to left atrium connection. AB - We report the case of an accessory pathway in a left inferoposterior diverticulum. The pathway masqueraded as a true left lateral pathway due to the direction of activation over a coronary sinus to left atrium connection. The patient had undergone four prior failed ablation attempts at other institutions using both a transseptal and retrograde approach. PMID- 12741714 TI - A newly characterized SCN5A mutation underlying Brugada syndrome unmasked by hyperthermia. AB - Febrile illness has been rarely reported to modulate ST segment elevation in right precordial leads on ECG or even precipitate ventricular fibrillation in patients with Brugada syndrome. We report the case of a patient whose Brugada ECG pattern was unmasked by hyperthermia secondary to acute cholangitis. Serial ECGs showed progressive attenuation of ST segment elevation as body temperature gradually returned to normal. Structural heart disease was ruled out. Intravenous flecainide injection reproduced a less remarkable ST segment elevation. Genetic screening demonstrated a single amino acid substitution (H681P) in the SCN5A gene, thus confirming the diagnosis of Brugada syndrome. In vitro expression of this newly characterized genetic defect revealed novel biophysical abnormalities consisting of a shift in both steady-state activation and inactivation, resulting in a 60% reduction of sodium window current. Thus, SCN5A-H681P mutation induces a significant loss of transmembrane current and is clinically associated with a pathologic phenotype that is elicited by hyperthermia. Overall the observed clinical features are in agreement with previous observations and strongly suggest that fever may be an environmental modifier among Brugada syndrome patients with a detrimental (and possibly arrhythmogenic) effect on cardiac repolarization. PMID- 12741715 TI - Relation between basic and clinical electrophysiologic characteristics in Brugada syndrome: facts or fiction? PMID- 12741716 TI - Useful tip to improve electrode positioning in markedly angulated coronary sinus tributaries. AB - The anatomic variability of the cardiac veins limits the feasibility of cardiac resynchronization therapy. This report describes another way to position the pacemaker electrode in sharply angulated coronary sinus branches. PMID- 12741717 TI - Left atrial flutter after radiofrequency catheter ablation of focal atrial fibrillation. AB - We report an arrhythmic complication in two patients in whom a procedure directed at isolating one or two pulmonary veins had been performed. The complication was related to pulmonary vein disconnection scars after ablation. Both patients developed new clinical tachycardia (atypical atrial flutter) secondary to a reentrant phenomena in the vicinity of a previously ablated pulmonary vein. PMID- 12741718 TI - Pericardial anatomy for the interventional electrophysiologist. AB - Investigators are beginning to exploit the pericardial space for a number of cardiovascular applications, including catheter ablation of cardiac arrhythmias, cardiovascular drug therapy, and cardiac pacing. This review explores the anatomy of the pericardial space and the anatomic variants that may be encountered in this novel approach to the heart. PMID- 12741719 TI - Inherited arrhythmic disorders in Japan. AB - The clinical and genetic characteristics of inherited arrhythmic disorders in Japan are briefly summarized. The incidence of hereditary long QT syndrome (LQTS) in Japan seems comparable to that in western countries. The genotypes are mainly LQT1 and LQT2; LQT3 and other types are rare. Mutations found in Japanese LQTS families are mostly novel compared to mutations reported in other countries and in different ethnic populations. Functional assays of the mutants in heterologous expression systems have disclosed novel mechanisms of current suppression in LQT1 and LQT2, and of gain of function in LQT3. Mutations in KCNJ2 may provide a new genotype (LQT7) of LQTS. In addition, mutations or single nucleotide polymorphisms in the channel genes responsible for LQTS (KvLQT1, HERG, and SCN5A) may predispose to drug-induced LQTS. A relatively high prevalence of Brugada syndrome is suspected in the Japanese population, and 1 of approximately 2,000 asymptomatic individuals present Brugada-type ECG changes upon annual examination. Genetic screening of the symptomatic Brugada syndrome and suspected cases has revealed SCN5A mutations in only approximately 12%. Therefore, the genetic basis of the majority of cases is not known. The expressed Na+ current of SCN5A mutant channels showed the phenotype of decreased channel function commonly seen in Brugada mutations. A case of idiopathic ventricular fibrillation was found to have a novel mutation in SCN5A, in which the expressed current showed marked suppression of channel function. PMID- 12741721 TI - Unusual phenomenon of spontaneous termination of atrioventricular nodal reentrant tachycardia with 2:1 atrioventricular block. PMID- 12741720 TI - Variable patterns of preexcitation: what is the mechanism? PMID- 12741722 TI - Termination of orthodromic supraventricular tachycardia with a nonpropagated stimulus. PMID- 12741723 TI - Pulmonary vein ablation using nonstandard electrodes: a plea for caution. PMID- 12741724 TI - International consensus on nomenclature and classification of atrial fibrillation: A collaborative project of the Working Group on Arrhythmias and the Working Group of Cardiac Pacing of the European Society of Cardiology and the North American Society of Pacing and Electrophysiology. PMID- 12741725 TI - Molecular mechanisms of fluoroquinolone resistance. AB - Fluoroquinolones have a broad spectrum of activity for complicated urinary tract infections, gastrointestinal infections, respiratory tract infections, sexually transmitted diseases, and chronic osteomyelitis. Since fluoroquinolones are excellent antibiotics for a number of clinical indications, their consumption has increased rapidly, both in human medicine and in food animals. Resistance to fluoroquinolones is chromosomal mediated, involving mutations either in the target genes including DNA gyrase (gyrA or gyrB) and topoisomerase IV (parC or parE), or in the regulatory factors controlling bacterial permeability or the efflux capacity of the bacteria. This review focuses on mechanisms of fluoroquinolone resistance, including known and proposed molecular mechanisms. This review also discuses the clinical impact of fluoroquinolone-resistant bacteria. PMID- 12741726 TI - Response to efavirenz plus two nucleoside reverse-transcriptase inhibitors in patients with advanced stage human immunodeficiency virus-1 infection in Taiwan. AB - From July 1, 1999 to April 30, 2002, 111 consecutive human immunodeficiency virus infected, antiretroviral-naive Taiwan patients initiated highly active antiretroviral therapy with efavirenz plus 2 nucleoside reverse-transcriptase inhibitors. Their median baseline CD4+ count was 50 x 10(6)/L (0-559 x 10(6)/L) and plasma viral load was 5.51 log10 copies/mL (3.09 to > 5.88 log10) as assessed by reverse-transcriptase polymerase chain reaction. Of the patients, 52.3% had a CD4+ count of < or = 50 x 10(6)/L, 74.8% had plasma viral load over 5 log10 copies/mL, and 58.5% had active AIDS-defining opportunistic illnesses. The median observation duration of antiretroviral therapy was 350 days (range, 28-991 days). At week 48 to 52 following the initiation of highly active antiretroviral therapy, 81. 8% (45/55) and 91.8% (45/49) of the patients achieved undetectable plasma viral load by intent-to-treat and ontreat analysis, respectively. At week 80 to 84, these percentage decreased to 69.7% (23/33) and 85.2% (23/27), respectively. Median CD4+ count increased from baseline to week 48 to 52 by 147 x 10(6)/L and to week 80 to 84 by 227 x 10(6)/L. The virologic and immunologic responses at each time period by intention-to-treat or on-treat analysis were similar between patients with baseline plasma viral load over or < or = 5 log10, CD4+ count over or < or = 50 x 10(6)/L, and with or without active AIDS-defining opportunistic illnesses. After initiation of highly active antiretroviral therapy for a median duration of 57 days (range, 2-638 days), 11 episodes of AIDS defining and 11 non-AIDS opportunistic illnesses occurred. The results of this study suggest that efavirenz plus 2 nucleoside reverse-transcriptase inhibitors is a potent antiretroviral combination regardless of whether the patient has a high baseline plasma viral load, low CD4+ count, or AIDS-defining opportunistic illnesses. PMID- 12741727 TI - Involvement of natural killer T cells in C57BL/6 mouse model of collagen-induced arthritis. AB - A subset of murine T lymphocytes sharing receptor structures with natural killer cells, named natural killer T cells, has characteristics distinct from conventional T cells and natural killer cells. The DBA/1 strain commonly used for collagen-induced arthritis induction does not express the natural killer 1.1 molecule, a marker for defining murine natural killer T cells. Therefore, collagen-induced arthritis was induced in the C57BL/6 strain carrying natural killer 1.1 marker and the role of natural killer T cells was examined in this model. The collagen-induced arthritis was induced successfully in the C57/BL6 strain with near 70% incidence. Lower percentages of natural killer T cells in lymphoid organs including spleen and lymph node and higher percentages of natural killer T cells in synovium were found in mice with severe inflamed joints as compared with those with mild inflamed joints. The results suggested an infiltration of natural killer 1.1+ CD3+ natural killer T cells into inflamed synovium in the model of collagen-induced arthritis. Although the preliminary results of natural killer 1.1 monoclonal antibody depletion experiments failed to alleviate the gravity of arthritis, such a C57BL/6 mouse model of collagen induced arthritis may provide a tool in the study of therapeutic manipulation of natural killerT cells in human autoimmune diseases such as rheumatoid arthritis. PMID- 12741728 TI - Predictive value of clinical features in differentiating group A beta-hemolytic streptococcal pharyngitis in children. AB - Identifying children with acute pharyngitis caused by group A beta-hemolytic Streptococcus (GABHS) is an important task for pediatricians. This study examined the value of certain clinical symptoms and signs in predicting a positive culture result. A total of 442 children who presented at the outpatient department with pharyngeal erythema were enrolled. The clinical features of patients with positive throat cultures for GABHS were compared to those with negative culture results. Throat cultures were positive for GABHS in 120 (27%) patients. Patients aged between 5 and 10 years had a higher prevalence of GABHS pharyngitis. Significant differences between the groups with and without GABHS pharyngitis were noted for the presence of sore throat (p < 0.001), tonsillar swelling (p < 0.001), anterior cervical adenopathy (p = 0.004), and scarlatiniform rash (p < 0.001), but not for the presence of fever, cough, rhinorrhea, abdominal pain, headache, tonsillar exudate, or palatal petechiae. Despite these strong associations, none of these symptoms or signs had both high sensitivity and specificity, and the positive predictive values of these individual findings were never greater than 50%. The results indicate that diagnosis based on clinical grounds alone is unreliable although there are certain individual symptoms and signs that are associated with GABHS pharyngitis. These symptoms and signs may be helpful in modifying estimates of probability of infection with GABHS. Throat cultures in suspected patients remain mandatory. PMID- 12741729 TI - Etiology of acute pharyngitis in children: is antibiotic therapy needed? AB - Acute pharyngitis is a common upper respiratory tract disease in children. The aim of this study is to find the associated microorganisms and determinate whether antibiotics is needed. This study included a total of 416 children with a diagnosis of acute pharyngitis who were treated in an outpatient clinic In Taipei. Throat swabs for viral and bacterial cultures were taken. Antibiotics were prescribed when bacterial pharyngitis was suspected on the initial visit. The prescription was adjusted according to the results of bacterial culture and clinical manifestations on the second visit 3 to 4 days later. The mean age of the patients was 52.9 +/- 36.9 months. A total of 297 potential pathogens were isolated in 242 patients. Viruses were isolated in 123 (29.6%) patients. Bacteria were isolated in 73 (17.5%) patients, whereas group A streptococci were isolated in only 7 (1.7%) patients. Viruses mixed with bacteria were found in 46 (11.1%) patients. The mean age of patients with viral infections was lower than those with bacterial infections (47.5 +/- 30.4 vs 62.4 +/- 43.7 months, p = 0.01). There was a longer duration of fever in patients older than 2 years with viral isolates (p < 0.01). Antibiotics were prescribed for acute pharyngitis on the first visit in 43 (10.3%) patients, and on the second visit in 19 (4.6%) patients. In children with viral infection, mixed isolates, or no growth, there was significantly less prescription of antibiotics on the second visit. Given the low isolation rate of significant bacterial pathogens, routine throat cultures and antibiotics are not indicated in children with acute pharyngitis. PMID- 12741730 TI - Prostatic abscess in southern Taiwan: another invasive infection caused predominantly by Klebsiella pneumoniae. AB - Prostatic abscess, though rarely encountered since the introduction of broad spectrum antibiotics, can cause significant morbidity and mortality. We retrospectively reviewed 17 cases of prostatic abscess treated during an 11-year period at 2 medical centers in southern Taiwan. Most of these patients were elderly (mean age, 59 years) with diabetes mellitus (10 cases, 59%) or hepatic cirrhosis (5 cases, 29%). Fourteen (82%) of the 17 patients were febrile, with chills occurring in about a half of these. Of the symptoms and signs referable to the lower urinary tract, dysuria (71%) was the most common complaint. Pain was usually localized in the suprapubic (35%) or perineal (18%) area. The common findings of digital rectal examination were prostatic enlargement (77%) and fluctuation (23%). Prostatic abscess was impressed from the findings of hypoechoic area with thick walls on transrectal ultrasound or an enlarged gland with fluid-density collections on computed tomography. All causative pathogens were gram-negative bacilli, including Klebsiella pneumoniae (10 cases), Escherichia coli (2), and Pseudomonas aeruginosa (1). Various measures were undertaken to allow drainage, including transurethral incision or resection of the prostate, open perineal incision, laparotomy, and transrectal ultrasound- or computed tomography-guided needle aspiration. In conclusion, K. pneumoniae was the predominant pathogen of prostatic abscess, and was frequently identified as the causative pathogen in patients with diabetes mellitus. Diagnosis of prostatic abscess based merely on symptomatology is implausible, and image studies, such as transrectal ultrasound or computed tomography scan, are warranted. Optimal management includes adequate drainage of abscess and antimicrobial therapy. PMID- 12741731 TI - Severe adenovirus infection in children. AB - During the period from July 1, 1999 to September 30, 2000, 9 children with severe adenovirus infection were treated at Chang Gung Children's Hospital. The mean age was 22 months (range, 5-50 months). All of them had lower respiratory tract infections, which manifested as lobar or segmental pneumonia and pleural effusion. Eight (88.9%) of the 9 patients required intensive care and 4 of them required mechanical ventilation. Abnormal laboratory findings included leukocytosis, elevated C-reactive protein, anemia, and prolonged prothrombin time and partial thromboplastin time. Extrapulmonary complications included hepatitis (6 cases), encephalitis (3), conjunctivitis (3), periorbital ecchymosis (1), and coagulopathy (2). One patient died, resulting in a mortality rate of 12.5%. Follow-up at 3 months postdischarge, 5 patients (62.5% of survivors) had bronchiolitis obliterans and/or organizing pneumonia. Seven patients were infected by serotype 3 adenovirus, 1 patient by serotype 2, and another by serotype 11. In conclusion, the clinical, laboratory, and radiographic features of severe adenovirus infection may mimic bacterial infection. Rapid progression of the clinical course despite antibiotic therapy and the presence of unusual extrapulmonary symptoms are important clinical clues in the diagnosis of severe adenoviral infection. PMID- 12741732 TI - Septic arthritis in children: relationship of causative pathogens, complications, and outcome. AB - This retrospective study investigated the causative pathogens, complications, and outcome of 58 children who were hospitalized for septic arthritis at a tertiary care hospital in southern Taiwan from July 1988 to December 2000. The mean age was 3 years (range, 12 days-16 years). The males/females ratio was 1.2:1. Ninety percent of the cases involved lower extremities (knee, hip, and ankle) with the hip being the most common site of infection (54%). Joint pain (81%) was the most common clinical presentation, followed by fever (74%), local warmness and swelling (72%), and limitation of motion (64%). Erythrocyte sedimentation rate was elevated (> or = 20 mm/h) initially in 89% of the cases. The predominant causative organism was Staphylococcus aureus (43%, 25/58), 6 isolates of which were methicillin-resistant, followed by coagulase-negative Streptococcus (6), Streptococcus pneumoniae (3), Salmonella spp. (3), Haemophilus influenzae type b (2), and group B Streptococcus (2). The concomitant complications of septic arthritis were sepsis (9%, 5/58) and meningitis (2%, 1/58). Ten patients had sequelae, including limitation of motion (6), limping gait (2), limb-length discrepancy (1), and abnormalities of bone growth (1). This study found that S. aureus was the most common infecting microorganism in septic arthritis in children. Septic arthritis with concomitant osteomyelitis and infection due to methicillin-resistant S. aureus was associated with a significantly increased risk of sequelae (relative risk, 46.4, 95% CI, 2.9-748.8; relative risk, 16. 2, 95% CI, 1.3-204.9, respectively). PMID- 12741733 TI - Early diagnosis of ventriculoperitoneal shunt infections and malfunctions in children with hydrocephalus. AB - This retrospective study assessed the parameters of ventriculoperitoneal shunt infection and malfunction found in 129 children with hydrocephalus who had undergone a ventriculoperitoneal shunt placement at Taipei Veterans General Hospital from January 1997 to June 2001. The clinical characteristics and differences among these patients were reviewed to determine the potential predictors of shunt infection and malfunction. Fever (60%, 6/ 10) and seizure (40%, 4/10) occurred significantly more often in the infection group than in the malfunction and control groups (p < 0.05). A higher blood C-reactive protein level was noted in the infection group than in the malfunction and control groups (p < 0.05). Examination of the cerebrospinal fluid of infected patients showed significantly higher white blood cell count and neutrophil count, higher protein concentration, and lower glucose levels compared with the other 2 groups (p < 0.05). However, the number of patients with cerebrospinal fluid eosinophilia was significantly higher in the malfunction group (p < 0.05). The diagnostic usefulness of laboratory parameters in patients with ventriculoperitoneal shunt infection is as follow: cerebrospinal fluid white blood cell count over 100/mm3, 96% specificity and a positive predictive value of 0.55; and cerebrospinal fluid neutrophils over 10%, 90% sensitivity and a negative predictive value of 0.99. In conclusion, Fever, seizure, high blood C-reactive protein, combined with leukocytosis, neutrophil over 10%, low glucose level, and high protein level in the ventricular fluid are factors that may help in distinguishing shunt infection from shunt malfunction. PMID- 12741734 TI - Coagulase-negative staphylococcal bacteremia in critically ill children: risk factors and antimicrobial susceptibility. AB - Coagulase-negative staphylococci (CoNS) are the most common microorganisms isolated from blood cultures in childern, and determining whether there is true bacteremia or merely contamination is a clinical dilemma. A total of 67 episodes of CoNS-positive blood cultures in pediatric and neonatal intensive care units were evaluated during a 3-year period in order to find the possible risk factors involved and the antimicrobial susceptibility of CoNS isolates. In this study, 37 episodes were judged to be infections as opposed to 30 that were not. In comparison with individuals without infection, patients with true infection of CoNS stayed longer in the hospital (32 +/- 32.9 vs 10.7 +/- 9.3 days, p = 0.001), had more surgical procedures (32.4% vs 6.7%, p = 0.014), received more antibiotic treatments in the recent 2 weeks (37.8% vs 0%, p < 0.001), underwent more central venous catheter insertions (86.4% vs 10%, p < 0.001), received more parenteral nutrition (37.8% vs 3.3%, p = 0.001), had higher C-reactive protein profiles (4.8 +/- 5.4 vs 0.6 +/- 0.9 mg/dL, p < 0.001), and had higher neutrophil proportion (58.1% vs 44.3%, p = 0.001). However, there were no significant differences in corticosteroid therapy, hemoglobin level, total leukocyte count, and platelet count. All strains of the infection group were resistant to cefazolin, cefotaxime, penicillin, and erythromycin. Nonetheless, all isolates were susceptible to vancomycin. The percentage of multiple-resistant CoNS in the infection group was 96.9%. Empirical therapy with vancomycin for CoNS bacteremia in critically ill children is therefore recommended. PMID- 12741735 TI - Is combination antimicrobial therapy required for urinary tract infection in children? AB - This retrospective study examined the characteristics of 338 pediatric patients presenting with a first episode of symptomatic urinary tract infection at Taichung Veterans General Hospital from November 1996 to December 2001. Escherichia coli was the most common pathogen (72.5%), followed by Proteus mirabilis (8.3%), Enterococcus (5.6%), and Klebsiella pneumoniae (4.7%). They were more susceptible to first-generation cephalosporin in comparison with other first-line antimicrobial agents such as trimethoprim/sulfamethoxazole, ampicillin, and gentamicin. Two hundred and eighty-seven (84.9%) of the 338 patients were divided into 3 groups according to the type of antibiotic treatment received, and the susceptibility rate and the averaged day of defervescence after effective antibiotic therapy were compared among the groups. Group 1 consisted of those patients treated with cefazolin or cephalexin alone (95%, 2.1 days); Group 2, cefazolin plus gentamicin (88.9%, 2.8 days); and Group 3, ampicillin plus gentamicin (76.1%, 2.3 days). A total of 38 (13.2%) cases from the 3 antibiotic groups did not respond to empiric antibiotics. For non-susceptible infections, when the antibiotic regimen was switched from cefazolin plus gentamicin to ampicillin alone, only 4 (20%) strains became susceptible, compared with 10 strains (62.5%) becoming susceptible after switching from ampicillin plus gentamicin to cefazolin alone (p < 0.01). The results indicated that first generation cephalosporin alone is an appropriate treatment for pediatric cases of community-acquired urinary tract infection and suggest that antimicrobial combinations should be reserved for serious or critical cases. PMID- 12741736 TI - Cyclosporin A therapy for steroid-dependent Henoch-Schonlein purpura. AB - Henoch-Schonlein purpura is one of the most common types of systemic vasculitis in children. Although recurrence is frequent, most cases are benign and self limited. Standard treatment consists of supportive care and nonsteroid antiinflammatory drugs and/or steroid. Here we report 2 cases of Henoch-Schonlein purpura, both of which had a prolonged and corticosteroid-dependent disease course. After treatment with cyclosporin A, the symptoms and signs subsided gradually in both cases with no recurrence after tapering of corticosteroid and cyclosporin A. PMID- 12741737 TI - Common variable immunodeficiency with hypoglycemia, Kikuchi lymphadenitis, and hemiparesis in two siblings. AB - Common variable immunodeficiency is a heterogeneous group of disorders with arthritis and/or arthralgia as its most commonly associated autoimmune manifestation. We report 2 cases of common variable immunodeficiency in siblings who also had other unusual signs and symptoms. A 10-year-old boy suffered from bradycardia, hypothermia, hypoglycemia, and chronic eczema. His 13-year-old sister suffered from Kikuchi lymphadenitis, hypoperfusion and atrophy of the left cerebral hemisphere, and hemiparesis. They both showed classical laboratory findings of common variable immunodeficiency and the boy's associated symptoms responded to intravenous immunoglobulin therapy. The findings from these cases suggest that a defect in the neuro-endocrine-immune axis may be one of the genetic bases of common variable immunodeficiency. PMID- 12741738 TI - Acute epiglottitis caused by Haemophilus influenzae type b: a case report. AB - Acute epiglottitis is an inflammatory, edematous disease of the epiglottis and adjacent structures, usually caused by Haemophilus influenzae type b. It is a life-threatening condition, occurring mainly in childhood. There have never been any reports of this condition in Taiwan. We report a case of 4-year-old boy who presented with characteristics of systemic illness combined with respiratory distress on arrival at the emergency room. His mouth was open and his neck was hyperextended. The diagnosis of epiglottitis was established on the basis of physical examination, lateral neck x-ray, and the finding of an enlarged, swollen, erythematous epiglottis on flexible fiberoptic laryngoscopy. Urine latex agglutination test for H. influenzae type b was positive and a blood culture grew H. influenzae type b. He was treated with cefotaxime and did not require intubation. PMID- 12741739 TI - Effects of permeabilizers on antimicrobial susceptibility of Stenotrophomonas maltophilia and Acinetobacter spp. AB - Isolates of the gram-negative bacteria Stenotrophomonas maltophilia and Acinetobacter spp. are naturally resistant to many classes of antibiotics. Using the disk diffusion technique it was shown that the membrane permeabilizers ethylenediaminetetraacetic acid, sodium citrate, and sodium polyphosphate increased susceptibility to a range of antibiotics including imipenem, ciprofloxacin, tetracycline, and rifampicin. These effects are probably due to the metal chelating properties of the permeabilizers. PMID- 12741740 TI - Partition priming in judgment under uncertainty. AB - We show that likelihood judgments are biased toward an ignorance-prior probability that assigns equal credence to each mutually exclusive event considered by the judge. The value of the ignorance prior depends crucially on how the set of possibilities (i.e., the state space) is subjectively partitioned by the judge. For instance, asking "what is the probability that Sunday will be hotter than any other day next week?" facilitates a two-fold case partition, [Sunday hotter, Sunday not hotter], thus priming an ignorance prior of 1/2. In contrast, asking "what is the probability that the hottest day of the week will be Sunday?" facilitates a seven-fold class partition, [Sunday hottest, Monday hottest, etc.], priming an ignorance prior of 1/7. In four studies, we observed systematic partition dependence: Judgments made by participants presented with either case or class formulations of the same query were biased toward the corresponding ignorance prior. PMID- 12741741 TI - Perceiving an object and its context in different cultures: a cultural look at new look. AB - In two studies, a newly devised test (framed-line test) was used to examine the hypothesis that individuals engaging in Asian cultures are more capable of incorporating contextual information and those engaging in North American cultures are more capable of ignoring contextual information. On each trial, participants were presented with a square frame, within which was printed a vertical line. Participants were then shown another square frame of the same or different size and asked to draw a line that was identical to the first line in either absolute length (absolute task) or proportion to the height of the surrounding frame (relative task). The results supported the hypothesis: Whereas Japanese were more accurate in the relative task, Americans were more accurate in the absolute task. Moreover, when engaging in another culture, individuals tended to show the cognitive characteristic common in the host culture. PMID- 12741742 TI - On the broad applicability of the affective circumplex: representations of affective knowledge among schizophrenia patients. AB - Studies of affective experience are guided by the assumption that the structure of affect generalizes across people. Yet this assumption has not been tested among educationally and economically diverse community residents or among individuals with psychopathology. This study explicitly examined the broad applicability of the valence-arousal circumplex and whether schizophrenia patients and nonpatients have comparable knowledge structures of affective phenomena. Patients and nonpatients completed similarity ratings of 120 pairs of affect words. Similarity judgments were analyzed separately for each group using a multidimensional scaling procedure, and solutions were compared. Results revealed the same two-dimensional valence-arousal solution for schizophrenia patients and nonpatients, although there were subtle differences between the groups. These findings provide additional evidence that the circumplex model is a useful formalism for representing affective phenomena across diverse populations, and they bolster confidence in existing interpretations of schizophrenia patients' reports of affective experience. PMID- 12741743 TI - IQ on the rise: the Flynn effect in rural Kenyan children. AB - Multiple studies have documented significant IQ gains over time, a phenomenon labeled the Flynn effect. Data from 20 industrialized nations show massive IQ gains over time, most notably in culturally reduced tests like the Raven's Progressive Matrices. To our knowledge, however, this is the first study to document the Flynn effect in a rural area of a developing country. Data for this project were collected during two large studies in Embu, Kenya, in 1984 and 1998. Results strongly support a Flynn effect over this 14-year period, with the most significant gains found in Raven's matrices. Previously hypothesized explanations (e.g., improved nutrition; increased environmental complexity; and family, parental, school, and methodological factors) for the Flynn effect are evaluated for their relevance in this community, and other potential factors are reviewed. The hypotheses that resonate best with our findings are those related to parents' literacy, family structure, and children's nutrition and health. PMID- 12741744 TI - Effects of experience on fetal voice recognition. AB - The ability of human fetuses to recognize their own mother's voice was examined. Sixty term fetuses were assigned to one of two conditions during which they were exposed to a tape recording of their mother or a female stranger reading a passage. Voice stimuli were delivered through a loudspeaker held approximately 10 cm above the maternal abdomen and played at an average of 95 dB SPL. Each condition consisted of three 2-min periods: no stimulus, voice (mother or stranger), and no stimulus. Fetal heart rate increased in response to the mother's voice and decreased in response to the stranger's; both responses were sustained for 4 min. The finding of differential behavior in response to a familiar versus a novel voice provides evidence that experience influences fetal voice processing. It supports an epigenetic model of speech perception, presuming an interaction between genetic expression of neural development and species specific experience. PMID- 12741745 TI - Transient spatial attention degrades temporal resolution. AB - To better understand the interplay between the temporal and spatial components of visual perception, we studied the effects of transient spatial attention on temporal resolution. Given that spatial attention sharpens spatial resolution, can it also affect temporal resolution? To assess temporal resolution, we measured the two-flash fusion threshold When two flashes of light are presented successively to the same location, the two-flash fusion threshold is the minimal interval between the flashes at which they are still perceived as two flashes, rather than a single flash. This assessment of temporal resolution was combined with peripheral precuing--a direct manipulation of transient spatial attention. This allowed us to demonstrate, for the first time, that spatial attention can indeed affect temporal resolution. However, in contrast to its effect on spatial resolution, spatial attention degrades temporal resolution. Two attentional mechanisms that could account for both attentional effects--enhanced spatial resolution and reduced temporal resolution--are discussed. PMID- 12741746 TI - Comparing the sensitivity of manual pursuit and perceptual judgments to pictorial depth effects. AB - We examined whether a pictorial depth illusion influences the manual pursuit of a moving dot to the same extent that it influences the dot's apparent displacement. Fourteen subjects performed two tasks. In one case, they used their unseen hand to track a dot that moved on an elliptical path. In the other, they first watched the dot move on the same path, and then set an ellipse to match the shape of the dot's path. The illusion influenced the two tasks to the same extent, suggesting that the visual information processing is the same for the two tasks. PMID- 12741747 TI - The development of numerical estimation: evidence for multiple representations of numerical quantity. AB - We examined children's and adults' numerical estimation and the representations that gave rise to their estimates. The results were inconsistent with two prominent models of numerical representation: the logarithmic-ruler model, which proposes that people of all ages possess a single, logarithmically spaced representation of numbers, and the accumulator model, which proposes that people of all ages represent numbers as linearly increasing magnitudes with scalar variability. Instead, the data indicated that individual children possess multiple numerical representations; that with increasing age and numerical experience, they rely on appropriate representations increasingly often; and that the numerical context influences their choice of representation. The results, obtained with second graders, fourth graders, sixth graders, and adults who performed two estimation tasks in two numerical contexts, strongly suggest that one cause of children's difficulties with estimation is reliance on logarithmic representations of numerical magnitudes in situations in which accurate estimation requires reliance on linear representations. PMID- 12741748 TI - Learning to recognize objects. AB - A theory of object recognition requires a theory of shape. Despite considerable empirical and theoretical research, however, a definition of object shape has proved elusive. Two experiments provide new insights by showing that children's object recognition changes dramatically during the period between 17 and 25 months. During this time, children develop the ability to recognize stylized three-dimensional caricatures of known and novel objects. This ability is linked to the number of object names in children's vocabularies, suggesting that category learning may be a driving force behind the developmental changes. PMID- 12741749 TI - Speech shadowing while driving: on the difficulty of splitting attention between eye and ear. AB - We investigated the role of cross-modal links in spatial attention in modulating the efficiency of dual-task performance. The difficulty of combining speech shadowing with a simulated driving task was modulated by the spatial location from which the speech was presented. In both single- and dual-task conditions, participants found it significantly easier to shadow one of two auditory streams when the relevant speech was presented from directly in front of them, rather than from the side. This frontal speech advantage was more pronounced when participants performed the demanding simulated driving task at the same time as shadowing than when they performed the shadowing task alone. These results demonstrate that people process auditory information more efficiently (with a lower overall dual-task decrement) when relevant auditory and visual stimuli are presented from the same, rather than different, spatial locations. These results are related to recent findings showing that there are extensive cross-modal links in spatial attention, and have clear implications for the design of better user interfaces. PMID- 12741750 TI - The undershoot bias: learning to act optimally under uncertainty. AB - Learning in stochastic environments is increasingly viewed as an important psychological ability. To extend these results from a perceptual to a motor domain, we tested whether participants could learn to solve a stochastic minimal time task using exploratory learning. The task involved moving a cursor on a computer screen to a target. We systematically varied the degree of random error in movement in three different conditions; each condition had a distinct time optimal solution. We found that participants approximated the optimal solutions with practice. The results show that adults are sensitive to the stochastic structure of a task and naturally adjust the magnitude of an undershoot bias to the particular movement error of a task. PMID- 12741751 TI - Good pitch memory is widespread. AB - Here we show that good pitch memory is widespread among adults with no musical training. We tested unselected college students on their memory for the pitch level of instrumental soundtracks from familiar television programs. Participants heard 5-s excerpts either at the original pitch level or shifted upward or downward by 1 or 2 semitones. They successfully identified the original pitch levels. Other participants who heard comparable excerpts from unfamiliar recordings could not do so. These findings reveal that ordinary listeners retain fine-grained information about pitch level over extended periods. Adults' reportedly poor memory for pitch is likely to be a by-product of their inability to name isolated pitches. PMID- 12741752 TI - Asymmetry in judgments of moral blame and praise: the role of perceived metadesires. AB - An important consideration in judging the blameworthiness (or praiseworthiness) of an action is whether the agent had sufficient control over it. In three experiments, we investigated judgments of moral blame and praise elicited when individuals were presented with vignettes describing actions that were performed either carefully and deliberately or impulsively and uncontrollably. Experiment 1 uncovered an asymmetry between judgments of positive versus negative actions- negative impulsive actions elicited a discounting of moral blame, but positive impulsive actions did not elicit a discounting of moral praise. Experiments 2 and 3 showed that this asymmetry arises because individuals judge agents on the basis of their metadesires (the degree to which the agents embrace or reject the impulses leading to their actions). Individuals assume that an agent would embrace an uncontrollable positive impulse, and reject an uncontrollable negative impulse. PMID- 12741753 TI - It does take a village: nonfamilial environments and children's behavior. AB - Family characteristics influence children's behavioral development, but so do variations in schools, neighborhoods, and communities. We documented extrafamilial environmental effects by fitting maximum likelihood models to questionnaire data collected from double dyads consisting of twins and their classmate controls. The classmate controls in each double dyad were genetic strangers living in separate households, but they shared school, neighborhood, and community environments with their yoked twin pair and with one another. At ages 11 to 12, the control classmates showed significant similarities in religious practices and smoking and drinking patterns, demonstrating that environmental influences outside the family affect children's behavioral development. Familial self-selection of residential neighborhoods may have contributed to these results, but direct effects of variation across communities, neighborhoods, and schools cannot be dismissed, and such effects warrant further study. PMID- 12741754 TI - Cognitive representation of negative numbers. AB - To understand negative numbers, must we refer to positive number representations (the phylogenetic hypothesis), or do we acquire a negative mental number line (the ontogenetic hypothesis)? In the experiment reported here, participants made lateralized button responses to indicate the larger of two digits from the range 9 to 9. Digit pairs were displayed spatially congruent or incongruent with either a phylogenetic or an ontogenetic mental number line. The pattern of decision latencies suggests that negative numbers become associated with left space, thus supporting the ontogenetic view. PMID- 12741755 TI - Children are cursed: an asymmetric bias in mental-state attribution. AB - Young children have problems reasoning about false beliefs. We suggest that this is at least partially the result of the same curse of knowledge that has been observed in adults--a tendency to be biased by one's own knowledge when assessing the knowledge of a more naive person. We tested 3- to 5-year-old children in a knowledge-attribution task and found that young children exhibited a curse-of knowledge bias to a greater extent than older children, a finding that is consistent with their greater difficulty with false-belief tasks. We also found that children's misattributions were asymmetric. They were limited to cases in which the children were more knowledgeable than the other person; misattributions did not occur when the children were more ignorant than the other person. This suggests that their difficulty is better characterized by the curse of knowledge than by more general egocentrism or rationality accounts. PMID- 12741756 TI - When prejudice does not pay: effects of interracial contact on executive function. AB - This study examined the influence of interracial interaction on the cognitive functioning of members of a dominant racial group. White participants had a brief interaction with either a White or a Black confederate, and then completed an ostensibly unrelated Stroop color-naming test. Prior to the interaction, participants' racial attitudes regarding Whites and Blacks were measured via the Implicit Association Test. Racial attitudes were predictive of impairment on the Stroop test for individuals who participated in interracial interactions, but not for those who participated in same-race interactions. The results are consistent with recently proposed resource models of self-regulation and executive control in that interracial interaction, a particularly taxing exercise of self regulation for highly prejudiced individuals, negatively affected performance on a subsequent, yet unrelated, test of executive function. PMID- 12741757 TI - Amplification of antioxidant activity and xanthine oxidase inhibition of catechin by enzymatic polymerization. AB - To amplify the antioxidant activity, we synthesized poly(catechin) by the enzyme catalyzed oxidative coupling using horseradish peroxidase as a catalyst. The poly(catechin) showed great improvement in antioxidant activity such as radical scavenging activity against the superoxide anion and inhibition effects against free radical induced-oxidation of low-density lipoprotein, compared with a catechin monomer. In addition, poly(catechin) showed very high inhibition effects on xanthine oxidase activity, whereas the catechin monomer showed very less inhibition effects. The amplified activities might offer a high potential as a therapeutic agent for prevention of various free radicals and/or enzyme-related diseases. PMID- 12741758 TI - Hybrid virus-polymer materials. 1. Synthesis and properties of PEG-decorated cowpea mosaic virus. AB - Cowpea mosaic virus was derivatized with poly(ethylene glycol) to give well controlled loadings of polymer on the outer surface of the coat protein assembly. The resulting conjugates displayed altered densities and immunogenicities, consistent with the known chemical and biological properties of PEG. These studies make CPMV potentially useful as a tailored vehicle for drug delivery. PMID- 12741759 TI - Synthesis of poly(L-lactide) end-capped with lactose residue. AB - The synthesis of poly(L-lactide) (polyLA) end-capped with lactose residue was studied from the standpoint of development of a new bioabsorbable material. After the hydroxyl group of t-butoxycarbonyl(Boc)-aminoethanol was converted to Boc aminoethanol-OK by using potassium/naphthalene, L-lactide was polymerized in tetrahydrofuran using Boc-aminoethanol-OK as an initiator at room temperature to prepare polyLA-NHBoc. Subsequently, the removal of the Boc group in terminal Boc aminoethanol residue was performed by treatment of formic acid to obtain the amino group end-capped polyLA (polyLA-NH(2)) as a reactive polyLA derivative. The coupling reactions of lactose with polyLA-NH(2) were investigated by two methods; the synthetic method through reductive amination of lactose with polyLA-NH(2) in the presence of sodium cyanoborohydride as a reducing agent did not give high degree of substitution of end-capped lactose residue per polyLA molecule, whereas the synthetic method through the ester interchange reaction of lactonolactone with polyLA-NH(2) gave Lac-polyLA perfectly end-capped with lactose residue. PMID- 12741760 TI - Fluorescent cellulose microfibrils as substrate for the detection of cellulase activity. AB - To devise a sensitive cellulase assay based on substrates having most of the physical characteristics of native cellulose, 5-(4,6 dichlorotriazinyl)aminofluorescein (DTAF) was used as a grafting agent to prepare suspensions of fluorescent microfibrils from bacterial cellulose. These suspensions were digested by a series of commercially relevant cellulases from Humicola insolens origin: cloned Cel6B and Cel 45A as well as crude H. insolens complex. The digestion induced the release of fluorescent cellodextrins as well as reducing sugars. After adequate centrifugation, these soluble products were analyzed as a function of grafting content, digestion time, and cellulase characteristics. The resulting data allowed the grafting conditions to be optimized in order to maximize the quantity of soluble products and therefore to increase the sensitivity of the detection. A comparison between the amount of released fluorescence and that of released reducing sugar allowed the differentiation between processive exo and endo cellulase activities. The casting of films of DTAF-grafted microfibrils at the bottom of the microwell titer plates also led to sensitive cellulase detection. As these films kept their integrity and remained firmly glued to the well bottom during the digestion time, they are tailored made for a full automation of the cellulases testing. PMID- 12741761 TI - Dissolution of Bombyx mori silk fibroin in the calcium nitrate tetrahydrate methanol system and aspects of wet spinning of fibroin solution. AB - There are still several problems associated with the spinning of dialyzed silk fibroin solutions. In this work some of these problems have been examined. The calcium nitrate tetrahydrate-methanol system was used to dissolve the silk fibroin. A compositional phase diagram was constructed at various concentrations of the solvent system. Regenerated fibroin powders from undialyzed fibroin solution in several coagulants showed different conformations. Regenerated powders from several coagulants except methanol and ethanol were resoluble in water. Atomic absorption analysis revealed that the calcium cations strongly interact with fibroin molecules in dialyzed fibroin solution, which may interfere with the regeneration of a strong fiber. Kinetic studies to determine the diffusion coefficient of methanol into dialyzed and concentrated fibroin solution were reported. The properties of both original and regenerated fibroin such as solubility in water and thermal behaviors using DSC were compared. Regenerated fibroin fiber was spun by the wet spinning method. An X-ray diffractogram showed that the regeneration process decreased the crystallinity of regenerated fibroin fiber. SEM images of the surface and cross section of the regenerated fibroin fibers were discussed. PMID- 12741762 TI - Synthesis and hydration properties of pH-sensitive p(HEMA)-based hydrogels containing 3-(trimethoxysilyl)propyl methacrylate. AB - An amphiphilic hydrogel network was synthesized from a cross-linked poly(2 hydroxyethyl methacrylate) backbone copolymerized with the monomers 3 (trimethoxysilyl)propyl methacrylate (PMA) and dimethylaminoethyl methacrylate (DMAEMA) using tetraethylene glycol diacrylate (TEGDA) as cross-linker and using the radical initiator system comprising N,N,N',N'-tetramethylethylenediamine and ammonium peroxydisulfate. The degree of hydration of hydrogel slabs was investigated as functions of varying monomer compositions and cross-link density and as a function of pH and ionic strength of the bathing medium. As much as a 45% increase in hydration was observed for hydrogels containing 15 mol % DMAEMA upon reducing the pH of the bathing medium from 8.0 to 2.0. This confirms the pH modulated swelling of amine-containing hydrogels. Increasing the concentration of TEGDA cross-linker from 3 to 12 mol % in a 10 mol % DMAEMA-containing hydrogel resulted in only a 10% reduction in the degree of hydration of the gel. There was, however, a 40-50% reduction in the degree of hydration of a 15 mol % DMAEMA hydrogel upon increasing the molar composition of PMA from 0 up to 20 mol %. The presence of PMA confers hydrophobic character that reduces hydration and introduces additional cross-links that reduce network mesh size. The water content of the hydrogel was consistently higher in buffers of lower ionic strength. The reversible pH-dependent swelling observed in these studies, along with the control of cross-link density afforded by the PMA component, endows these biocompatible materials with potential for use in pH-controlled drug delivery of more hydrophobic drugs and present new compositions for in vitro and in vivo biocompatibility studies. PMID- 12741763 TI - Mechanism of the polymerization reaction initiated and catalyzed by the polyhydroxybutyrate synthase of Ralstonia eutropha. AB - Polyhydroxybutyrate (PHB) synthases (polymerases) catalyze the polymerization of the coenzyme A thioester of 3-hydroxybutyrate to PHB. The Ralstonia eutropha PHB synthase purified from recombinant E. coli cells exists in aqueous solution in both monomeric (single subunit) and homodimeric (two subunits) forms in equilibrium. Several lines of evidence suggest that the homodimer is the active form of the synthase. The initial mechanistic model for the polymerization reaction proposed that two different thiol groups form the catalytic site. The cysteine at 319 has been shown to provide one thiol group that is involved in the covalent catalysis, but a second thiol group on the same protein molecule has not yet been identified. It is suggested that cysteines at 319 from each of the two molecules of a homodimer synthase provide two identical thiol groups to jointly form a single catalytic site. To verify this model using the strategy of in vitro reconstitution, heterodimers composed of the wild-type subunit and of the C(319) mutated subunit were constructed and the activities at various ratios of the wild type subunit to the mutated subunit were measured. The experimental results indicate that the homodimer is the active form of the enzyme, that the heterodimer containing the mutated subunit has no activity, and that a single cysteine is not sufficient for catalysis. Two identical thiol groups from C(319) residues on each subunit of the homodimer are required to form the catalytic site for the initiation and propagation reactions. We further demonstrate that a dimer synthase that has initiated the polymerization reaction (primed synthase) is significantly more stable against dissociation than the unprimed (unreacted) dimer synthase. These two properties explain the nature of lag phenomenon during the in vitro polymerization reaction catalyzed by this enzyme PMID- 12741764 TI - Imaging of anisotropic cellulose suspensions using environmental scanning electron microscopy. AB - The effect of concentration on anisotropic phase behavior of acid-hydrolyzed cellulose suspensions has been examined using conventional polarizing microscopy and the novel technique of environmental scanning electron microscopy (ESEM). Microcrystalline cellulose dispersed in water formed biphasic suspensions in a narrow concentration range, 4-12 wt % for a suspension pH of 4, where the upper and lower phases were isotropic and anisotropic (chiral nematic), respectively. It is known from previous work that within the biphasic regime total suspension concentration affects only the volume fractions of the two phases, not phase concentration or interfacial packing. As the total suspension concentration surpassed the upper critical limit (c), however, a single anisotropic phase of increasing concentration was observed. It was evident from polarizing microscopy that the chiral nematic pitch of the anisotropic phase decreased with increasing concentration, which has been attributed to a reduction in the electrostatic double layer thickness of the individual rods, thus increasing intermolecular interactions. Chiral nematic textures were also visible using ESEM. This technique has the advantage of studying individual rod orientation within the liquid crystalline phase as it permits the high resolution of electron microscopy to be applied to hydrated samples in their natural state. To our knowledge this is the first time such lyotropic systems have been observed using electron microscopy. PMID- 12741765 TI - Characterization of peptide-amphiphiles possessing cellular activation sequences. AB - Numerous approaches have been described for modifying biomaterials to incorporate extracellular matrix components. "Peptide-amphiphiles", whereby monoalkyl hydrocarbon chains are covalently linked to peptide sequences, have been shown previously to (a) form specific molecular architecture with enhanced stability and (b) promote cell adhesion, spreading, and signaling. The present study has examined the use of chimeric peptide-amphiphiles for inducing protein-like structures and peptide-amphiphile mixtures for enhancing surface bioactivity. The alpha-helical propensity of a 21 residue peptide, incorporating the SPARC(119 122) angiogenesis-inducing sequence and either unmodified or acylated with a C(6), C(10), C(14), C(16), C(18), C(18:1), or C(18:1-OH) monoalkyl hydrocarbon chain, has been examined. Peptide and peptide-amphiphile structures were characterized by circular dichroism and one- and two-dimensional NMR spectroscopic techniques. The 21 residue peptide alone does not form a distinct structure in solution, whereas N-terminal acylation by monoalkyl hydrocarbon chains results in the 21 residue peptide-amphiphile adopting a predominantly alpha-helical structure in solution. The thermal stability of the alpha-helix increases with increasing hydrocarbon chain length. The SPARC(119-122) peptide amphiphiles were then screened for promotion of endothelial cell adhesion and spreading. The greatest activity was achieved by using a mixture of the alpha helical SPARC(119-122) peptide-amphiphile, a triple-helical peptide-amphiphile incorporating the alpha2beta1 integrin binding site from type I collagen, and a pseudolipid. The pseudolipid is most likely required for a spatial distribution of the peptide-amphiphiles that allows for optimal cellular interactions. Overall, we have found that incorporation of bioactive sequences within peptide amphiphiles results in the induction of an ordered structure of the bioactive sequence and that mixtures of peptide-amphiphiles can be used to promote endothelial cell behaviors comparable to extracellular matrix components. PMID- 12741766 TI - Long-term release and improved intracellular penetration of oligonucleotide polyethylenimine complexes entrapped in biodegradable microspheres. AB - The aim of this work was to design a biodegradable delivery system for oligonucleotides providing both a sustained release and an improved intracellular penetration. To this purpose oligonucleotide/polyethylenimine (ON/PEI) complexes at nitrogen to phosphate (N/P) molar ratios of about 15 or 40 were encapsulated into poly(lactide-co-glycolide) microspheres by the multiple emulsion-solvent evaporation technique. ON/PEI complexes were efficiently entrapped inside microspheres. The introduction of salts within the external aqueous phase allowed an improvement of microsphere characteristics. In particular, the use of sodium chloride led to a reduced microsphere porosity and a more homogeneous ON distribution inside the polymeric matrix. These effects were attributed to the reduced flux of water from the external aqueous phase toward the internal aqueous droplets, due to the osmotic effect of sodium chloride. Both, the reduced porosity and the improved ON distribution inside the matrix, were considered responsible for the lower burst effect and the slower ON release rate from microsphere prepared with sodium chloride. ON/PEI complexes encapsulated inside microspheres were also protected toward enzymatic degradation in fetal calf serum. Interestingly, ON/PEI complexes slowly released from microspheres efficiently penetrated inside HeLa cells and oligonucleotides were preferentially located in the nucleus. PMID- 12741768 TI - Lipase-catalyzed polycondensations: effect of substrates and solvent on chain formation, dispersity, and end-group structure. AB - The effects of substrates and solvent on polymer formation, number-average molecular weight (M(n)), polydispersity, and end-group structure for lipase catalyzed polycondensations were investigated. Diphenyl ether was found to be the preferred solvent for the polyesterification of adipic acid and 1,8-octanediol giving a M(n) of 28 500 (48 h, 70 degrees C). The effect of varying the alkylene chain length of diols and diacids on the molecular weight distribution and the polymer end-group structure was assessed. A series of diacids (succinic, glutaric, adipic, and sebacic acid) and diols (1,4-butanediol, 1,6-hexanediol, and 1,8-octanediol) were polymerized in solution and in bulk. It was found that reactions involving monomers having longer alkylene chain lengths of diacids (sebacic and adipic acid) and diols (1,8-octanediol and 1,6-hexanediol) give a higher reactivity than reactions of shorter chain-length diacids (succinic and glutaric acid) and 1,4-butanediol. The bulk lipase-catalyzed condensation reactions were feasible, but the use of diphenyl ether gave higher M(n) values (42,400 g/mol in 3 days at 70 degrees C). The polydispersity varied little over the conditions studied giving values 200 degrees C) and residence time, resulted in lower mean molecular weights and yield. Oligosaccharides with higher mean molecular weights were obtained at lower severity, but the yield was considerably lower. The feasibility of using the extracted hemicellulosic oligosaccharides from spruce for the synthesis of hydrogels was demonstrated. PMID- 12741778 TI - Galactose-substituted alginate: preliminary characterization and study of gelling properties. AB - Coupling of alginate with 1-amino-1-deoxygalactose in the presence of 1-ethyl-3 [3-(dimethylamino)propyl]carbodiimide results in a substituted polymer containing galactose side linked via an amide bond. To clarify the degree and pattern of substitution, a (1)H NMR study on the anomeric region of modified alginate, polymannuronate, alginate enriched in guluronic acid (G-enriched alginate), and polyalternating MG, was carried out (G, alpha-l-guluronic acid; M, beta-d mannuronic acid). From the resonance of the proton at position 1 of galactosylamine, it was possible to determine the amount of galactose linked to mannuronic and to guluronic residues, respectively. Furthermore, (1)H NMR spectroscopy revealed a higher reactivity of guluronic residues for low degrees of conversion. Modified alginates with 7% and 19% of substitution are both able to form stable beads in the presence of calcium ions. The effect of galactose substitution on the dimensions, swelling, and stability of the beads has been studied and the cytotoxicity of the modified polymer evaluated in preliminary biological tests. PMID- 12741779 TI - Effect of oxidation rate on cross-linking of mussel adhesive proteins. AB - The cross-linking behavior of mussel adhesive protein Mefp-1 was studied by measuring the rate of aggregation of the protein by photon correlation spectroscopy. To be able to calculate the aggregation numbers, the hydrodynamic radius of monomer Mefp-1 (10 nm) was determined under reducing conditions. The aggregation is controlled by the redox potential of the solution, and the aggregation number varied, independent of pH, over a factor 2 within the experimentally accessible redox potential window. A kinetic model for cross linking, based on the intricate interplay of the oxidation and auto-oxidation of the hydroquinones of Mefp-1, is proposed. The oxidation rate strongly depends on redox potential. The cross-linking rate is taken to be proportional to the rate of auto-oxidation. The model correctly predicts the experimentally observed phenomena. When the oxidation rate is slower than the auto-oxidation rate, cross linking is efficient and controlled by the oxidation rate. When the rate of auto oxidation rate is slower than the oxidation rate, the cross-linking is inefficient due to the quick exhaustion of the hydroquinones. The experimentally determined rate constant for cross-linking is found to be much smaller than those found for auto-oxidation of hydroquinones because of the excluded volume interactions imposed by the protein backbone. Tuning the interplay between oxidation and auto-oxidation presents the potential of controlling cross-linking density independent of the density of reactive groups. PMID- 12741780 TI - Typical physicochemical behaviors of chitosan in aqueous solution. AB - Physicochemical properties of a homogeneous series of chitosans with different degrees of acetylation and almost the same degree of polymerization were investigated in an ammonium acetate buffer. Techniques such as interferometry, static light scattering (in batch or coupled on line with a chromatographic system), and viscometry were processed. All of the results agree with a unique law of behavior only depending on the degree of acetylation of the polymer. Indeed, values of the refractive index increment, radius of gyration, second viral coefficient, and intrinsic viscosity are decreasing in the same way as DA is increasing. Three distinct domains of DA were defined and correlated to the different behaviors of chitosans: (i) a polyeletrolyte domain for DA below 20%; (ii) a transition domain between DA = 20% and 50% where chitosan loses its hydrophilicity; (iii) a hydrophobic domain for DAs over 50% where polymer associations can arise. Conformations of chitosan chains were studied by the calculations of the persistence lengths (L(p)). The average value was found to be close to 5 nm, in agreement with the wormlike chain model, but no significant variation of L(p) with the degree of acetylation was noticed. PMID- 12741781 TI - A keystone-like grain in hornet comb cells: its nature and physical properties. AB - In the roof of cells built by the Oriental hornet, Vespa orientalis (Vespinae, Hymenoptera), there is one or more grains differing in their composition and shape from the surrounding structures. These grains have a diameter of 100-200 microm and protrude slightly from the roof inward. They contain primarily a large percentage of Ti but also Fe and occasionally Si. The mineral makeup in the grains is still uncertain, but it seems that the minerals belong to the group called perovskite. These grains in the hornet's comb may originate from the soil around the nest, that is, comprised of ceramic matter, but it is also possible that they are secreted from a gland within the frons plate of the hornet because the fungus-like secretion in this gland contains elements similar to those in the roof of the comb cells. We conjecture that the grain in question serves as a reference point for the hornets that build the cell walls in the direction of gravity. The cement whereby the building hornets conjoin the various building materials, whether organic (cellulose and the like) or inorganic (sand grains or tiny stones), is a saliva that is secreted from their oral gland, which upon release hardens into a polymeric glue. The ceramic grain embedded within this polymeric matrix is endowed with different physical properties and can therefore respond differently to the tension or pressure of the surrounding walls. PMID- 12741782 TI - Crab shell chitin whisker reinforced natural rubber nanocomposites. 1. Processing and swelling behavior. AB - Nanocomposite materials were obtained from a colloidal suspension of chitin whiskers as the reinforcing phase and latex of both unvulcanized and prevulcanized natural rubber as the matrix. The chitin whiskers, prepared by acid hydrolysis of chitin from crab shell, consisted of slender parallelepiped rods with an aspect ratio close to 16. After the two aqueous suspensions were mixed and strirred, solid composite films were obtained either by freeze-drying and hot pressing or by casting and evaporating the preparations. The processing and swelling behavior of composite films were evaluated. It was concluded that the whiskers form a rigid network assumed to be governed by a percolation mechanism in the evaporated samples only. Comparatively, better resistance of evaporated samples than hot-pressed ones against swelling in an organic solvent medium is good evidence for the existence of a rigid chitin network. The values of diffusion coefficient, bound rubber content, and relative weight loss also supported the presence of a three-dimensional chitin network within the evaporated samples. The mechanical behavior of the composites gives additional insight and evidence for this fact (part 2). PMID- 12741784 TI - Enzyme-containing Michael-adduct-based coatings. AB - A two-step method was developed to homogeneously insert carbonic anhydrase (CA, E.C. 4.2.1.1) into Michael-adduct-based coatings. CA was first covalently coupled to an N-vinylformamide-based water-soluble polymer. Unlike native CA, the resulting polymer/CA system could be dispersed within a film matrix. The enzyme containing coating (ECC) hydrolyzes p-nitrophenyl propionate in buffered media at high rates retaining approximately 7% apparent activity. In comparison, other two step techniques for the chemical coupling of CA to the coating surface were less efficient and led to coatings with significantly less activity. A three-step immobilization process coupling the enzyme to the surface of a partially hydrolyzed coating also raised retention of activity after coating synthesis. CA ECC is stable under ambient conditions retaining 45% activity after 90 days of storage at room temperature. PMID- 12741783 TI - Crab shell chitin whisker reinforced natural rubber nanocomposites. 2. Mechanical behavior. AB - In a previous work (part 1), nanocomposite materials were obtained using a latex of either unvulcanized or prevulcanized natural rubber as the matrix and a colloidal suspension of crab chitin whiskers as the reinforcing phase. The mechanical behavior of the resulting nanocomposite films was analyzed in both the linear and the nonlinear range in the present study. The effects of the filler and processing technique were evaluated, and the results are discussed based on the knowledge of the structural morphology and swelling behavior reported in our previous work. The reinforcing effect of chitin whiskers strongly depended on their ability to form a rigid three-dimensional network, resulting from strong interactions such as hydrogen bonds between the whiskers. The results emanating from the successive tensile test experiments give clear evidence for the presence of a three-dimensional chitin network within the evaporated samples. Cross linking of the matrix was found to interfere with the formation of this network. PMID- 12741786 TI - Tailoring the swelling pressure of degrading dextran hydroxyethyl methacrylate hydrogels. AB - Swelling pressure measurements were performed on degrading dextran hydroxyethyl methacrylate (dex-HEMA) hydrogels. In these networks, the cross-links are hydrolyzable carbonate ester bonds formed between methacrylate groups and dextran molecules. It is demonstrated that dex-HEMA gels made in the presence of a known amount of free dextran chains exhibit osmotic properties similar to those of partially degraded dex-HEMA gels. The swelling pressure, Pi(sw), of degrading dex HEMA gels is controlled primarily by the cross-linked dex-HEMA polymer and the free dextran molecules, while the contribution of short poly-HEMA fragments (produced in the degradation process) is negligible. It is found that Pi(sw) only slightly changes during the first 15 days of degradation. Close to the end of the degradation process, however, a much faster increase in Pi(sw) is observed. The swelling pressure profile of these gels strongly depends on the concentration of the cross-linked dex-HEMA and its chemical composition (amount of HEMA groups per 100 glucose units). PMID- 12741785 TI - Effect of polymer ionization on the interaction with DNA in nonviral gene delivery systems. AB - The optimization of DNA-cationic polymer complexation is crucial for nonviral gene delivery. Although physicochemical characterization of the interaction between DNA and cationic polymers has recently attracted more attention in the nonviral DNA delivery field, the literature on the effect of varying polycation charge density on DNA-cationic polymer complexation is still scarce. Thus, the aim of this study was to systematically assess the influence of the degree of ionization of a weak cationic polyelectrolyte (poly[2-(dimethylamino)ethyl methacrylate] or DMAEMA homopolymer) on its ability to form complexes with DNA. This was achieved by varying the solution pH from 4.0 to 8.0 and analyzing the resulting effects on the binding affinity, thermodynamic properties, complex size, and morphology. Lowering the solution pH led to higher degrees of ionization for the cationic polymer and hence greater binding affinities with DNA, as judged by the increased propensity of the former to displace ethidium bromide from DNA and also by relatively low monomer:nucleotide molar ratio (0.8:1) required to retard the migration of free DNA. Isothermal titration microcalorimetry studies further confirmed that a stronger interaction occurred at low pH than at high pH. By decreasing the pH from 8.0 to 6.6, K(obs) increased from 7.8 x 10(5) to 20.4 x 10(5) M(-1). More efficient condensation at low pH was demonstrated by the reduction of ethidium bromide fluorescence in the loading wells from gel electrophoresis, decreased complex sizes without agglomeration occurring at high polymer/DNA ratios, together with discrete and dense spherical complexes observed in TEM studies. This may be attributed to the presence of electrostatic stabilization from excess cationic polymer chains, which provide a repulsive shell around the polymer/DNA complex. The physicochemical data indicate that the increased degree of ionization for the DMAEMA homopolymer at lower pH results in higher binding affinity, smaller and more compact complexes, and more efficient condensation. These findings therefore highlight the importance of the degree of ionization on DNA complex formation for weak cationic polyelectrolytes. PMID- 12741787 TI - Impact of silicone-based block copolymer surfactants on the surface and bulk microscopic organization of a biodegradable polymer, poly(epsilon-caprolactone). AB - Two amphiphilic AB block copolymers, containing a highly compatible poly(epsilon caprolactone) (PCL) block connected to a poly(dimethylsiloxane) (PDMS) block having a low surface energy, are synthesized and characterized in terms of their dispersion in a presynthesized PCL matrix. X-ray photoelectron spectroscopy, contact angle measurements, atomic force microscopy, and optical microscopy are used to describe the evolution of the surface chemical composition, as well as the surface and bulk morphology of the PCL/copolymer blends as a function of the nature and weight surface free energy and the dispersion of the copolymers in the blends, leading to important modifications of the bulk and the surface morphology. These differences are interpreted in terms of the impact of the block copolymers on the semicrystalline polymer structure and related properties in the prospect of using the surfactants to improve the synthesis of PCL in supercritical CO(2). PMID- 12741788 TI - Water-soluble degradable hyperbranched polyesters: novel candidates for drug delivery? AB - A novel approach to hyperbranched polymers is presented in this work. Hyperbranched polyesters with a large amount of terminal hydroxyl groups are prepared by a one-pot synthesis from commercially available AB-type and CD(n) type monomers (n >/= 2). In this paper, Michael addition of diethanolamine (CD(2)) or N-methyl-d-glucamine (CD(5)) to methyl acrylate (AB) generates dominantly AD(n)-type intermediates. Further self-condensation of intermediates at higher temperature and in the presence of catalyst gives hyperbranched polyesters. Because of the tertiary amino groups in the backbone and the hydroxyl groups in the linear and terminal units, the resulting hyperbranched polyester is highly soluble in water. Furthermore, the hyperbranched polymer is degradable because of its ester units. So, the water-soluble hyperbranched polyesters might be applied as a novel material for drug delivery. PMID- 12741789 TI - Synthesis and physicochemical characterization of end-linked poly(ethylene glycol)-co-peptide hydrogels formed by Michael-type addition. AB - The synthesis of novel hybrid hydrogels by stepwise copolymerization of multiarm vinyl sulfone-terminated poly(ethylene glycol) macromers and alpha-omega cysteine oligopeptides via Michael-type additions is described. Cross-linking kinetics, studied by in situ rheometry, can be controlled by pH and the presence of charged amino acid residues in close proximity to the Cys, which modulates the pK(a) of the thiol group. These end-linked networks were characterized by their equilibrium swelling in water, by their viscoelastic properties in the swollen state, and by their soluble fraction. It was demonstrated that structure and properties are very sensitive to the preparation state including stoichiometry and precursor concentration and less sensitive to the pH during cross-linking. For each network the concentration of elastically active chains (nu) was calculated from experimentally determined sol fractions using Miller-Macosko theory and compared to values obtained from swelling and rheometry studies and by calculation from Flory's classical network models. Hydrogels were also prepared with varying macromer structures, and their properties were shown to respond to both macromer functionality and molecular weight. PMID- 12741790 TI - A Mossbauer study of the interaction of chitosan and D-glucosamine with iron and its relevance to other metalloenzymes. AB - The interaction of iron with water-soluble polymer chitosan and monomer d glucosamine is investigated by Mossbauer spectroscopy. The 4.2 K Mossbauer spectrum of Fe-water-soluble chitosan complex indicates the presence of a magnetic pattern and a quadrupole doublet, and analysis of the spectral data leads to the conclusion that an Fe(II) state is partially stabilized in this system. Fe-glucosamine (monomer of chitosan) complex, on the other hand, clearly stabilizes the Fe(II) state in the acidic pH range as evidenced from the isomer shift extracted from the Mossbauer spectra. The oxidation state of the metal ion in the complex is found to be pH dependent. Indirect evidence supporting the involvement of amino group in the bonding with the metal ion is discussed. From the analysis of the experimental data under varying experimental conditions, it is concluded that the metal ion in the complex is at least tetracoordinated and at most hexacoordinated with O/N ligands of the polymer or monomer and thus corroborates the bonding scheme proposed earlier. PMID- 12741791 TI - Intramolecular complex formation of poly(N-isopropylacrylamide) with human serum albumin. AB - Complexation of human serum albumin (HSA) with poly(N-isopropylacrylamide) (PNIPA) ranging in molecular weight (M(PNIPA)) from 2.1 x 10(4) to 1.72 x 10(6) was studied in an aqueous system (pH 3) containing NaCl as a supporting salt. Dynamic light scattering, static light scattering, electrophoretic light scattering, and dialyzing techniques were used as the experimental tool in a suitable combination. The measurements were performed mainly at 25 degrees C and at 0.01 M NaCl as a function of mixing ratio (r(m), molar ratio of PNIPA to HSA). The results of DLS and ELS evidently demonstrated the formation of a water soluble complex through mixing of HSA and PNIPA. A detailed analysis of SLS data with the aid of dialysis data revealed that the resulting complex is an "intramolecular" complex consisting of a PNIPA chain with several of bound HSA molecules. Both hydrodynamic radius (R(h)) and radius gyration (R(g)) of intramolecular complexes decreased as r(m) was increased. This result correlated well to the fact that the number (n) of bound proteins per polymer decreases with increasing r(m). The size and the molar mass of the complex became large depending on M(PNIPA), but the increase of M(PNIPA) led to a decrease in n at r(m) < 1. The increase in NaCl concentration from 0.01 to 0.3 M brought about the increase in the size and the molar mass of an intramolecular HSA-PNIPA complex prepared at r(m) = 1.1. This was found to be due to an increase of n. A similar trend was observed when temperature rose from 25 to 32 degrees C (close to lower critical solution temperature of PNIPA). However, the effect of temperature on the increase of was strong in comparison with that of ionic strength. On the basis of these results obtained, the complexation mechanism was discussed in detail. PMID- 12741792 TI - Thermally induced conformation change of succinoglycan in aqueous sodium chloride. AB - Static and dynamic light scattering, viscosity, and optical rotation measurements have been made at eight different temperatures between 25 and 75 degrees C on two succinoglycan samples (sodium salt) with weight-average molecular weights M(w) of 7.14 x 10(5) and 3.54 x 10(5) (at 25 degrees C) in 0.01 M aqueous NaCl to investigate the thermally induced order-disorder conformation change of the polysaccharide. Additionally, viscometry and polarimetry have been performed for a sodium salt sample (M(w) = 4.55 x 10(5) at 25 degrees C) whose M(w), z-average radius of gyration (z)(1/2), and hydrodynamic radius R(H) in the aqueous salt had been determined previously. As the temperature increases, M(w), (z)(1/2), R(H), and the intrinsic viscosity for every sample sharply decrease around 55 degrees C where the specific rotation at 300 nm sigmoidally increases. In particular, M(w) at 25 degrees C (i.e., in the ordered helical state) is twice as large as that at 75 degrees C (i.e., in the disordered state). These findings substantiate that the ordered structure is composed of two chains and hence is a double helix. Data analysis shows that this helix at 25 degrees C is characterized by an unperturbed wormlike chain with a helix pitch of about 2 nm (per repeating unit) and a persistence length of about 50 nm and that upon heating, it dissociates directly (i.e., in all-or-none fashion) to disordered chains of a similar contour length but with a much smaller persistence length of about 10 nm. The temperature dependence of the light scattering second viral coefficient is discussed in relation to the association of disordered chains in the cooling process. PMID- 12741793 TI - A novel method for the determination of carbonyl groups in cellulosics by fluorescence labeling. 3. Monitoring oxidative processes. AB - The fluorescence-based CCOA method for determination of carbonyl group profiles in cellulosic substrates was employed to study the mechanisms of various oxidative and degradation processes involving celluloses in greater detail. The approach comprises labeling with the marker carbazole-9-carboxylic acid [2-(2 aminooxyethoxy)ethoxy]amide (CCOA), followed by gel permeation chromatography in DMAc/LiCl with fluorescence, multiangle laser light scattering, and refractive index detection. At first, the CCOA method was applied to study solutions of pulp in N-methylmorpholine-N-oxide monohydrate (NMMO), as occurring in the production of Lyocell-type fibers. NMMO is a rather strong oxidant that on one hand converts reducing end groups to carboxyl structures, thus lowering the overall carbonyl content, but generates new keto structures along the chain by nonselective oxidation on the other hand. The CCOA method allowed for the first time to distinguish the carbonyl course in different molecular weight ranges. Second, alkalization and aging of pulp, which are used in the industrial preparation of cellulose derivatives, e.g., as an element of the preripening process in viscose rayon production, were investigated. The CCOA method shows a clear reduction of the molecular weight, accompanied by a fast loss of carbonyls in the first phase, which is due to removal of low-molecular weight material by dissolution, and a slow decrease in the second phase, which is caused by further oxidation of carbonyl groups. Also here, differences in the carbonyl course in different molecular weight regions were monitored. Third, electron beaming, proposed as a means of pulp activation, was shown to decrease and narrow the molecular weight distribution, under generation of comparatively low amounts of carbonyls, which, however, are also introduced into high molecular weight, crystalline domains, as shown by a comparison of homogeneous and heterogeneous CCOA labeling approach. Finally, as the fourth application, thermal treatment of cellulose at temperatures between 105 and 165 degrees C was shown to bring about a small reduction of the molecular weight, which only at higher drying temperatures is accompanied by an introduction of carbonyls over the whole molecular weight range. PMID- 12741795 TI - Temperature-responsive cellulose by ceric(IV) ion-initiated graft copolymerization of N-isopropylacrylamide. AB - Temperature-responsive cellulose has been obtained by graft copolymerization of N isopropylacrylamide (NIPAAm) monomer using ceric ammonium nitrate (CAN) as initiator at 25.0 +/- 0.1 degrees C in acidic medium. Kinetic and grafting parameters were evaluated at different concentrations of NIPAAm ranging from 1.25 x 10(-3) to 12.5 x 10(-3) mol dm(-3) and varying concentrations of CAN from 1.5 x 10(-3) to 9.0 x 10(-3) mol dm(-3) at constant concentration of nitric acid (2.5 x 10(-2) mol dm(-3)). The graft copolymerization of NIPAAm onto cellulose has shown a significant increasing trend below lower critical solution temperature (LCST) of poly(N-isopropylacrylamide) (PNIPAAm) and shown low energy of activation (18.0 kJ mol(-1)) for graft copolymerization within the temperature range of 10-35 degrees C as determined with Arrhenius plot. The PNIPAAm-grafted cellulose has shown improved thermal stability and shown temperature-dependent degree of swelling. Variation in degree of swelling of PNIPAAm-grafted cellulose as a function of temperature has been used to determine LCST of PNIPAAm-grafted cellulose. The contact angle (theta) has shown variation on increasing the graft yield and temperature. On the basis of experimental observations, the reaction steps for graft copolymerization have been proposed and a rate expression has been derived. PMID- 12741794 TI - Amphotericin B encapsulated in micelles based on poly(ethylene oxide)-block poly(L-amino acid) derivatives exerts reduced in vitro hemolysis but maintains potent in vivo antifungal activity. AB - The core-forming blocks of amphiphilic diblock copolymers based on methoxypoly(ethylene oxide)-block-poly(L-aspartate), PEO-b-p(L-Asp), were derivatized to incorporate stearate side chains. The effects of stearate esterification were assessed in terms of micelle stability and amphotericin B (AmB) encapsulation/release. The level of stearate esterification modulates the relative self-aggregation state of encapsulated AmB as evidenced by absorption spectroscopy. When AmB is physically loaded into polymeric micelles, the onset of hemolytic activity toward bovine erythrocytes is delayed relative to that of the free drug. Furthermore, the extent of esterification (0, 46, or 91%) appears to have profound influence on the time-dependent hemolytic profile of AmB toward bovine erythrocytes. Particularly in the case of highly substituted stearate ester micelles, incomplete and gradual build-up of hemolysis was observed over a period of 24 h. On the basis of the corresponding absorption spectra, we speculate that encapsulated AmB may interact strongly with stearate side chains, resulting in sustained release. In a neutropenic murine model of disseminated candidiasis, kidney colony-forming unit determination revealed dose-dependent efficacy for the polymeric micelle/AmB formulation, which was not significantly different from that of Fungizone at doses of 0.2, 0.3, and 0.6 mg/kg (p = 0.7). Thus, AmB administered via a polymeric micelle formulation retained potent in vivo activity. PMID- 12741796 TI - Alternative intracellular signaling mechanism involved in the inhibitory biological response of functionalized PMMA-based polymers. AB - A PMMA-based polymer previously shown to inhibit cell proliferation was compared to untreated PMMA. Conformation of adsorbed proteins, cell adhesion, cytoskeleton formation, and integrin activation were examined. Fibronectin adsorbed in a different conformation on the PMMA-based polymer exposing a different balance of the heparin-binding domains. Fibroblasts attached in equal numbers to both surfaces over a 4-h period, but the integrins involved in the adhesion process elicited different intracellular signaling pathways. Cells attached to PMMA showed activation of FAK and MAP as they spread using an assembled actin cytoskeleton. Cells attached to the polymer showed early and strong MAP activity that resulted in nonassembly of the actin cytoskeleton and sub-optimal cell spreading. We conclude that the chemistry of the polymer surface dictated a different conformation of the adsorbed proteins that resulted in alternative cell signaling and diminished cell spreading. This accounted for the biological inhibition previously reported on the PMMA-based polymer. PMID- 12741797 TI - Epoxy-amino groups: a new tool for improved immobilization of proteins by the epoxy method. AB - The properties of a new commercially available amino-epoxy support (amino-epoxy Sepabeads) for immobilizing enzymes have been compared to those of conventional epoxy supports. The new support has a layer of epoxy groups over a layer of ethylenediamine that is covalently bound to the support. Thus, this support has a great anionic exchanger power and a high number of epoxy groups. We have found a number of advantages to this new heterofunctional support. Immobilization proceeds at low ionic strength using amino epoxy Sepabeads while requiring high ionic strength using conventional monofunctional epoxy supports. Immobilization is much more rapid using amino-epoxy supports than employing conventional epoxy supports. The possibility of achieving immobilized preparations in which the enzyme orientation may be different to that obtained using the traditional hydrophobic supports (with likely effects in terms of activity or stability). Stability of the immobilized enzyme has been found to be much higher using the new support than in preparations using the conventional ones in many cases. Here we show some examples of these advantages using different enzymes (beta galactosidases, lipase, glutaryl acylase, invertase, and glucoamylase). PMID- 12741798 TI - Uptake of atmospheric carbon dioxide into silk fiber by silkworms. AB - The relation between the uptake of atmospheric CO(2) and insect's production of silk fiber has not yet been reported. Here, we provide the first quantitative demonstrations that four species of silkworms (Bombyx mori, Samia cynthia ricini, Antheraea pernyi, and Antheraea yamamai) and a silk-producing spider (Nephila clavata) incorporate atmospheric CO(2) into their silk fibers. The abundance of (13)C incorporated from the environment was determined by mass spectrometry and (13)C NMR measurements. Atmospheric CO(2) was incorporated into the silk fibers in the carbonyl groups of alanine, aspartic acid, serine, and glycine and the C(gamma) of aspartic acid. We show a simple model for the uptake of atmospheric CO(2) by silkworms. These results will demonstrate that silkworm has incorporated atmospheric CO(2) into silk fiber via the TCA cycle; however, the magnitude of uptake into the silk fibers is smaller than that consumed by the photosynthesis in trees and coral reefs. PMID- 12741799 TI - Melting and crystallization behaviors of biodegradable polymers enzymatically coalesced from their cyclodextrin inclusion complexes. AB - Inclusion complexed (IC) and coalesced biodegradable poly(epsilon-caprolactone) (PCL), poly(L-lactic acid) (PLLA), and their diblock copolymer (PCL-b-PLLA) were achieved by forming ICs between host alpha-cyclodextrin(alpha-CD) and guest PCL, PLLA, and PCL-b-PLLA, followed by removing the alpha-CD host with an amylase enzyme. FTIR spectra of the coalesced polymers reveal that the host alpha-CD can be completely removed, without polymer degradation, by treatment with an amylase enzyme. The melting and crystallization behavior of these CD-IC treated polymers, which are crystallizable, biodegradable, and bioabsorbable, are investigated by differential scanning calorimetry (DSC) and polarized optical microscopy. Results show that coalescence increased the crystallinities of the homopolymers but decreased that of the diblock copolymer. The Avrami exponent (n), derived from both isothermal and nonisothermal crystallization models for homo-PCL and -PLLA and the PCL and PLLA blocks in the diblock copolymer samples coalesced from their ICs, is close to 4, indicating homogeneous crystallization, whereas crystallization of the blocks in the as-synthesized diblock copolymer yields an Avrami exponent around 3, indicating heterogeneous crystallization. All of these results demonstrate that the PCL and PLLA homopolymers and blocks in the IC coalesced samples are more readily and homogeneously crystallized than those in the as-synthesized samples or their physical blend, even though the level of crystallinity in the IC-coalesced diblock copolymer is significantly lower. Moreover, unlike the as-synthesized diblock copolymer, the crystallization of PCL and PLLA blocks in the IC-coalesced diblock copolymer are not influenced by their covalent connection. PMID- 12741800 TI - Injectable polymeric microspheres with X-ray visibility. Preparation, properties, and potential utility as new traceable bulking agents. AB - The copolymer of methyl methacrylate (MMA) and 2-[2',3',5' triiodobenzoyl]oxoethyl methacrylate (1), ratio 3:1 (mass:mass), was prepared via a free-radical polymerization in bulk. The copolymer (M(w) = 97.8 kD and M(n) = 41.5 kD) was dissolved in chloroform and subsequently transformed into beads with a diameter in the micrometer range, using a solvent evaporation technique. The resulting microbeads were characterized by different techniques, including NMR spectroscopy, differential scanning calorimetry, gel permeation chromatography, and scanning electron microscopy. The latter technique was used as the basis for statistical analysis of the bead size. Typically, an average diameter of 96 microm and a standard deviation of 21 microm were obtained. The beads were also subjected to some preliminary tests regarding cytotoxicity. The copolymer of MMA and 1 contains covalently bound iodine. Therefore, the material is intrinsically radiopaque, i.e., capable of absorbing X-radiation while no contrast additive is needed. Our interest in these microspheres stems primarily from their possible utility as injectable and afterward traceable (radiopaque) bulking agents, e.g., for use in urology for the treatment of female stress incontinence due to sphincter deficiency. As a first test into this direction, a sample of the microbeads was mixed with ethylene glycol, and the resulting suspension was studied with respect to injectability and radiopacity. The results suggest that the radiopaque microbeads may provide access to improved bulking agents. Further modification of the surface may be necessary in order to suppress the migratory aptitude of the radiopaque polymeric microspheres in vivo. PMID- 12741801 TI - Pressure cell assisted solubilization of xyloglucans: tamarind seed polysaccharide and detarium gum. AB - To improve the solubilization of two water-soluble xyloglucans, tamarind seed polysaccharide and detarium gum, by reducing substantially molecular aggregation, a "pressure cell" heating method was used. Conditions allowing solubilization and chain depolymerization were produced by varying appropriately the pressure, time, and temperature applied. The various MW fractions of solubilized xyloglucans were characterized by capillary viscometry and light scattering techniques in order to extract, with reliability, fundamental macromolecular parameters. Mark-Houwink and Flory exponents were found to be 0.67 +/- 0.04 and 0.51 +/- 0.06, respectively for both xyloglucan data combined, consistent with linear random coil behavior. A detailed analysis of the data seems to suggest that tamarind gum solutions are slightly perturbed by the effect of excluded volume, whereas detarium gum samples are close to the theta state. Chain flexibility parameters such characteristic ratio, C( proportional, variant ), and persistence length, L(p), were calculated for tamarind and detarium using the Burchard-Stockmayer Fixman (BSF) geometric method. L(p) values of 6-8 nm were estimated for xyloglucans. The seemingly linear structure of tamarind and detarium, as suggested by the value of the Mark-Houwink and Flory exponents obtained, follows from analysis of the data by the classical Zimm method but not when employing the square root or Berry method which suggests a more branched chain profile. This was the approach adopted in our previous work on the characterization of detarium samples. PMID- 12741802 TI - Freeze-fracture electron microscopy of lipid membranes on colloidal polyelectrolyte multilayer coated supports. AB - Lipid membranes were assembled on polyelectrolyte (PE)-coated colloidal particles. The assembly was studied by means of confocal microscopy, flow cytometry, scanning force microscopy, and freeze-fracture electron microscopy. A homogeneous lipid coverage was established within the limits of optical resolution. Flow cytometry showed that the lipid coverage was uniform. Freeze fracture electron microscopy revealed that the lipid was adsorbed as a bilayer, which closely followed the surface profile of the polyelectrolyte support. Additional adsorption of polyelectrolyte layers on top of the lipid bilayer introduced inhomogeneities as evident from jumps in the fracture plane. Characteristic lipid multilayers have not been seen with freeze-fracture electron microscopy. PMID- 12741804 TI - Heterogeneous acylation of flax fibers. Reaction kinetics and surface properties. AB - Flax fibers composed mainly of cellulose were subjected to heterogeneous valerylation reaction. The progress of the chemical modification was assessed by transmission FTIR. The heterogeneous esterification reaction followed first-order kinetics, and a plateau was reached already after 30 min. The intensity of the FTIR hydroxyl absorption band (nu = 3400 cm(-1)) did not appreciably decrease during the acylation reaction, showing that only a small fraction of the fiber hydroxyls was involved in the reaction. The degree of valerate substitution (DS) at the fiber surface (50 A thick layer) was evaluated by means of ESCA. Surface valerylation increased with reaction time and leveled off at DS around 1 after 30 min, in agreement with the FTIR data. The chemically modified fibers maintain the Cellulose I crystal structure and the original crystallinity degree up to the longest reaction time investigated (180 min). Dynamic contact angle measurements showed that surface hydrophobicity as indicated by advancing contact angle rapidly increased upon valerylation reaching a plateau after about 10 min. Chemical modification does not appreciably alter fiber thermal stability (by TGA) and morphology (by SEM). PMID- 12741803 TI - Synthesis and characterization of chimeric silkworm silk. AB - A synthetic gene encoding a chimeric silklike protein was constructed that combined a polyalanine encoding region (Ala)(18), a sequence slightly longer than the (Ala)(12-13) found in the silk fibroin from the wild silkworm Samia cynthia ricini, and a sequence encoding GVGAGYGAGAGYGVGAGYGAGVGYGAGAGY, found in the silk fibroin from the silkworm Bombyx mori. A tetramer of the chimeric repeat sequence encoding a approximately 29 kDa protein was expressed as a fusion protein in Escherichia coli. In comparison to S. c. ricini silk, the chimeric protein demonstrated improved solubility because it could be dissolved in 8 M urea. The purified protein assumed an alpha-helical structure based on solid-state (13)C CP/MAS NMR and was less prone to conformational transition to a beta-sheet, unlike native silk proteins from S. c. ricini. Model peptides representing the crystalline region of S. c. ricini silk fibroin, (Ala)(12) and (Ala)(18), formed beta-sheet structures. Therefore, the solubility and structural transitions of the chimeric protein were significantly altered through the formation of this chimeric silk. This experimental strategy to the study of silk structure and function can be used to develop an improved understanding of the contributions of protein domains in repetitive silkworm and spider silk sequences to structure development and structural transitions. PMID- 12741805 TI - Synthesis and properties of star-shaped polylactide attached to poly(amidoamine) dendrimer. AB - Star-shaped polylactide was synthesized by bulk polymerization of lactide with poly(amidoamine) (PAMAM) dendrimer as initiator, which was marked as PAMAM-g-PLA for simplicity. The nonlinear architecture of PAMAM-g-PLA was confirmed by gel permeation chromatograph, nuclear magnetic resonance, and differential scanning calorimetry analysis. Unlike the linear polylactide (PLA) with similar molecular weight, PAMAM-g-PLA had a higher hydrophilicity and a faster degradation rate because of shortened polymer chains and increased polar terminal endgroups due to its branch structure. The highly branched structure significantly accelerated the release of water-soluble bovine serum albumin from PAMAM-g-PLA microspheres, whereas the linear PLA with similar molecular weight exhibited an initial time lag release. This star polymer may have potential applications for hydrophilic drug delivery in tissue engineering, including growth factor and antibodies to induce tissue regeneration, by adjusting the chain lengths of PLA branches. PMID- 12741806 TI - A new strategy for recycling and preparation of poly(L-lactic acid): hydrolysis in the melt. AB - Poly(L-lactide) [i.e., poly(L-lactic acid) (PLLA)] was hydrolyzed in the melt in high-temperature and high-pressure water at the temperature range of 180-350 degrees C for a period of 30 min, and formation, racemization, and decomposition of lactic acids and molecular weight change of PLLA were investigated. The highest maximum yield of l-lactic acid, ca. 90%, was attained at 250 degrees C in the hydrolysis periods of 10-20 min. Too-high hydrolysis temperatures such as 350 degrees C induce the dramatic racemization and decomposition of formed lactic acids, resulting in decreased maximum yield of L-lactic acid. The hydrolysis of PLLA proceeds homogeneously and randomly via a bulk erosion mechanism. The molecular weight of PLLA decreased exponentially without formation of low molecular-weight specific peaks originating from crystalline residues. The activation energy for the hydrolysis (deltaE(h)) of PLLA in the melt (180-250 degrees C) was 12.2 kcal x mol(-1), which is lower than 20.0 kcal x mol(-1) for PLLA and 19.9 kcal x mol(-1) for poly(dl-lactide) [i.e., poly(DL-lactic acid)] as a solid in the temperature range below the glass-transition temperature (21-45 degrees C). This study reveals that hydrolysis of PLLA in the melt is an effective and simple method to obtain l-lactic acid and to prepare PLLA having different molecular weights without containing the specific low-molecular-weight chains, because of the removal of the effect caused by crystalline residues. PMID- 12741807 TI - Planar supported bilayer polymers formed from bis-diene lipids by Langmuir Blodgett deposition and UV irradiation. AB - Substrate-supported lipid bilayers have been prepared from bis-diene functionalized phosphorylcholine (PC) lipids and polymerized by UV irradiation. The overall bilayer structure is largely preserved upon removal from water, although significant loss of material occurs from the upper leaflet of the bilayer, likely due to desorption at the air/water interface. The morphology and surface structure of the bilayer, as observed by AFM, indicate a substantially different arrangement of the lipids in the hydrated and dehydrated states, presumably due to the loss of water from the near surface region. These changes have been correlated with infrared spectral shifts sensitive to the conformation of the hydrocarbon chains. Protein adsorption studies show that rehydrated, polymerized bilayers retain a degree of resistance to BSA adsorption intermediate between model hydrophobic and fluid PC lipid bilayer surfaces. The degree of protein adsorption is correlated with desorption of material from the upper leaflet of the bilayer upon drying, which produces voids at which hydrophobically driven protein adsorption occurs. PMID- 12741808 TI - Directed capture of enzymes and bacteria on bioplastic films. AB - The development of smart coatings for a variety of uses depends on the ability of the coating material to perform specific functions. We have used water dispersible polyurethane preparations for the immobilization of binding proteins under mild conditions. In these experiments, antibodies against the enzyme beta galactosidase or the bacterium Escherichia coli were immobilized in polyurethane coatings and then used to effectively capture their cognate antigen. Further, a second, more general, capture protocol was developed which involves the incorporation of the protein avidin in the plastics. This system efficiently captures biotinylated beta-galactosidase. Biotinylated anti-E. coli antibody captured by avidin bioplastics resulted in a nearly 5-fold increase in the number of bound bacteria when compared to blank polyurethane. The use of avidin in a bioplastic allows any biotinylated antibody to be applied to all or part of the surface resulting in a patterning of capture agents on a preformed surface. PMID- 12741809 TI - Self-assembly of poly([R]-3-hydroxybutyric acid)-block-poly(ethylene glycol) diblock copolymers. PMID- 12741810 TI - Substrate-induced conformational changes in Escherichia coli taurine/alpha ketoglutarate dioxygenase and insight into the oligomeric structure. AB - The enzymes in the alpha-ketoglutarate (alphaKG) dependent dioxygenase superfamily represent the largest class of non-heme iron oxidases and have important medical, ecological, and biotechnological roles. One such enzyme, taurine/alpha-ketoglutarate dioxygenase (TauD), catalyzes the conversion of 2 aminoethanesulfonate (taurine) to sulfite and aminoacetaldehyde while decomposing alphaKG to succinate and CO(2). This alphaKG dependent dioxygenase is expressed in Escherichia coli under sulfur starvation conditions and allows the cell to utilize taurine, and other similar sulfonates in the environment, as an alternative sulfur source. In this work, we report the structures of the apo and holo forms of TauD to 1.9 A resolution (R(cryst) = 21.2%, R(free) = 24.9%) and 2.5 A resolution (R(cryst) = 22.5%, R(free) = 27.8%), respectively. The models reported herein provide significant new insight into the substrate orientations at the active site and the conformational changes that are induced upon taurine binding. Furthermore, analysis of our crystallographic data coupled with reanalysis of the crystallographic model (resolution = 3.0 A, R(cryst) = 28.1, R(free) = 32.0) presented by Elkins et al. (Biochemistry (2002) 41, 5185-5192) reveals an alternative oligomeric arrangement for the enzyme that is consistent with the conserved primary and secondary structure elements of other alphaKG dependent dioxygenases. PMID- 12741811 TI - Insights into the evolution of allosteric properties. The NADH binding site of hexameric type II citrate synthases. AB - Study of the hexameric and allosterically regulated citrate synthases (type II CS) provides a rare opportunity to gain not only an understanding of a novel allosteric mechanism but also insight into how such properties can evolve from an unregulated structural platform (the dimeric type I CS). To address both of these issues, we have determined the structure of the complex of NADH (a negative allosteric effector) with the F383A variant of type II Escherichia coli CS. This variant was chosen because its kinetics indicate it is primarily in the T or inactive allosteric conformation, the state that strongly binds to NADH. Our structural analyses show that the six NADH binding sites in the hexameric CS complex are located at the interfaces between dimer units such that most of each site is formed by one subunit, but a number of key residues are drawn from the adjacent dimer. This arrangement of interactions serves to explain why NADH allosteric regulation is a feature only of hexameric type II CS. Surprisingly, in both the wild-type enzyme and the NADH complex, the two subunits of each dimer within the hexameric conformation are similar but not identical in structure, and therefore, while the general characteristics of NADH binding interactions are similar in each subunit, the details of these are somewhat different between subunits. Detailed examination of the observed NADH binding sites indicates that both direct charged interactions and the overall cationic nature of the sites are likely responsible for the ability of these sites to discriminate between NADH and NAD(+). A particularly novel characteristic of the complex is the horseshoe conformation assumed by NADH, which is strikingly different from the extended conformation found in its complexes with most proteins. Sequence homology studies suggest that this approach to binding NADH may arise out of the evolutionary need to add an allosteric regulatory function to the base CS structure. Comparisons of the amino acid sequences of known type II CS enzymes, from different Gram negative bacteria taxonomic groups, show that the NADH-binding residues identified in our structure are strongly conserved, while hexameric CS molecules that are insensitive to NADH have undergone key changes in the sequence of this part of the protein. PMID- 12741812 TI - Capturing enzyme structure prior to reaction initiation: tropinone reductase-II substrate complexes. AB - To understand the catalytic mechanism of an enzyme, it is crucial to determine the crystallographic structures corresponding to the individual reaction steps. Here, we report two crystal structures of enzyme-substrate complexes prior to reaction initiation: tropinone reductase-II (TR-II)-NADPH and TR-II-NADPH tropinone complexes, determined from the identical crystals. A combination of two kinetic crystallographic techniques, a continuous flow of the substrates and Laue diffraction measurements, enabled us to capture the transit structures prior to the reaction proceeding. A structure comparison of the enzyme-substrate complex elucidated in this study with the enzyme-product complex in our previous study indicates that one of the substrates, tropinone, is rotated relative to the product so as to make the spatial organization in the active site favorable for the reaction to proceed. Side chains of the residues in the active site also alter their conformations to keep the complementarity of the space for the substrate or the product and to assist the rotational movement. PMID- 12741813 TI - Crystal structure of a catalytic site mutant of beta-amylase from Bacillus cereus var. mycoides cocrystallized with maltopentaose. AB - The X-ray crystal structure of a catalytic site mutant of beta-amylase, E172A (Glu172 --> Ala), from Bacillus cereus var. mycoides complexed with a substrate, maltopentaose (G5), and the wild-type enzyme complexed with maltose were determined at 2.1 and 2.0 A resolution, respectively. Clear and continuous density corresponding to G5 was observed in the active site of E172A, and thus, the substrate, G5, was not hydrolyzed. All glucose residues adopted a relaxed (4)C(1) conformation, and the conformation of the maltose unit for Glc2 and Glc3 was much different from those of other maltose units, where each glucose residue of G5 is named Glc1-Glc5 (Glc1 is at the nonreducing end). A water molecule was observed 3.3 A from the C1 atom of Glc2, and 3.0 A apart from the OE1 atom of Glu367 which acts as a general base. In the wild-type enzyme-maltose complex, two maltose molecules bind at subsites -2 and -1 and at subsites +1 and +2 in tandem. The conformation of the maltose molecules was similar to that of the condensation product of soybean beta-amylase, but differed from that of G5 in E172A. When the substrate flips between Glc2 and Glc3, the conformational energy of the maltose unit was calculated to be 20 kcal/mol higher than that of the cis conformation by MM3. We suggest that beta-amylase destabilizes the bond that is to be broken in the ES complex, decreasing the activation energy, DeltaG(++), which is the difference in free energy between this state and the transition state. PMID- 12741814 TI - Solution structure, dynamics, and thermodynamics of the native state ensemble of the Sem-5 C-terminal SH3 domain. AB - Although the high-resolution structure of a protein may provide significant insight into which regions are important for function, it is well-known that proteins undergo significant conformational fluctuations, even under native conditions. This suggests that the static structure alone may not provide sufficient information for elucidation of the thermodynamic determinants of biological function and that an accurate molecular-level description of function requires knowledge of the nature and energetics of the conformational states that constitute the native state ensemble. Here the native state ensemble of the C terminal src homology domain-3 (C-SH3) from Caenorhabditis elegans Sem-5 has been studied using a variety of high-resolution biophysical techniques. In addition to determining the first solution structure of the unliganded protein, we have performed (15)N relaxation and native state hydrogen-deuterium exchange. It is observed that the regions of greatest structural variabilility also show low protection and order parameters, suggesting a higher degree of conformational diversity. These flexible regions also coincide with those regions of Sem-5 that have been predicted by the COREX algorithm to be unfolded in many of the most probable conformational states within the native state ensemble. The implications of this agreement and the potential role of conformational heterogeneity of the observed biophysical properties are discussed. PMID- 12741815 TI - Structures along the catalytic pathway of PrmC/HemK, an N5-glutamine AdoMet dependent methyltransferase. AB - Posttranslational methylation of release factors on the glutamine residue of a conserved GGQ motif is required for efficient termination of protein synthesis. This methylation is performed by an N(5)-glutamine methyltransferase called PrmC/HemK, whose crystal structure we report here at 2.2 A resolution. The electron density at the active site appears to contain a mixture of the substrates, S-adenosyl-L-methionine (AdoMet) and glutamine, and the products, S adenosyl-L-homocysteine (AdoHcy) and N(5)-methylglutamine. The C-terminal domain of PrmC adopts the canonical AdoMet-dependent methyltransferase fold and shares structural similarity with the nucleotide N-methyltransferases in the active site, including use of a conserved (D/N)PPY motif to select and position the glutamine substrate. Residues of the PrmC (197)NPPY(200) motif form hydrogen bonds that position the planar Gln side chain such that the lone-pair electrons on the nitrogen nucleophile are oriented toward the methyl group of AdoMet. In the product complex, the methyl group remains pointing toward the sulfur, consistent with either an sp(3)-hybridized, positively charged Gln nitrogen, or a neutral sp(2)-hybridized nitrogen in a strained conformation. Due to steric overlap within the active site, proton loss and formation of the neutral planar methylamide product are likely to occur during or after product release. These structures, therefore, represent intermediates along the catalytic pathway of PrmC and show how the (D/N)PPY motif can be used to select a wide variety substrates. PMID- 12741816 TI - High-resolution crystal structures and spectroscopy of native and compound I cytochrome c peroxidase. AB - Cytochrome c peroxidase (CCP) is a 32.5 kDa mitochondrial intermembrane space heme peroxidase from Saccharomyces cerevisiae that reduces H(2)O(2) to 2H(2)O by oxidizing two molecules of cytochrome c (cyt c). Here we compare the 1.2 A native structure (CCP) with the 1.3 A structure of its stable oxidized reaction intermediate, Compound I (CCP1). In addition, crystals were analyzed by UV-vis absorption and electron paramagnetic resonance spectroscopies before and after data collection to determine the state of the Fe(IV) center and the cationic Trp191 radical formed in Compound I. The results show that X-ray exposure does not lead to reduction of Fe(IV) and only partial reduction of the Trp radical. A comparison of the two structures reveals subtle but important conformational changes that aid in the stabilization of the Trp191 cationic radical in Compound I. The higher-resolution data also enable a more accurate determination of changes in heme parameters. Most importantly, when one goes from resting state Fe(III) to Compound I, the His-Fe bond distance increases, the iron moves into the porphyrin plane leading to shorter pyrrole N-Fe bonds, and the Fe(IV)-O bond distance is 1.87 A, suggesting a single Fe(IV)-O bond and not the generally accepted double bond. PMID- 12741817 TI - Identification of the residues in the helix F/G loop important to catalytic function of membrane-bound prostacyclin synthase. AB - A topological model of prostaglandin I(2) synthase (PGIS) was created by homology modeling. This model, along with site-specific antibodies and other topology studies, has suggested that the residue(s) within helix F/G loop of PGIS may be involved in forming the substrate access channel and located in a position that influences the membrane-bound PGIS catalytic function (1). To test this hypothesis, we have explored an approach to identify the residues of the helix F/G loop important to enzyme activity of the membrane-bound PGIS by a combination of 2-D NMR experiment and mutagenesis methods. Using the distance measured from the model as a guide, the helix F/G loop was mimicked in a synthetic peptide by introducing a spacer to maintain a distance of about 7 A between the N- and the C termini (PGIS residues 208 and 230). The peptide was used to interact with the enzyme substrate analogue, U46619. High-resolution 2-D NMR experiments were performed to determine the contacts between the peptide and U46619. The interaction between the constrained F/G loop peptide and U46619 was confirmed by the observation of the conformational changes of the peptide and U46619 using the comparison of the cross-peaks between the NOESY spectra of U46619 with the peptide, without the peptide, and the peptide alone. Through the combination of the 2-D NMR experiments, completed (1)H NMR assignments of the F/G loop segment in the presence and absence of U46619 were obtained, and these data were used to predict the contact residues (Leu214 and Pro215) of the F/G loop with PGIS substrate. The predicted influence of residues on enzyme catalytic activity in membrane-bound environments was confirmed by the mutagenesis of the F/G loop residues of human PGIS. These observations support that the F/G loop is involved in forming the substrate access channel for membrane-bound PGIS and suggests that the NMR experiment-based mutagenesis approach may be applied to study structure and function relationships for other proteins. PMID- 12741818 TI - Key amino acid residues in the regulation of soluble methane monooxygenase catalysis by component B. AB - The regulatory component MMOB of soluble methane monooxygenase (sMMO) has been hypothesized to control access of substrates into the active site of the hydroxylase component (MMOH) through formation of a size specific channel or region of increased structural flexibility tuned to methane and O(2). Accordingly, a decrease in the size of four MMOB residues (N107G/S109A/S110A/T111A, the Quad mutant) was shown to accelerate the reaction of substrates larger than methane with the reactive MMOH intermediate Q [Wallar, B. J., and Lipscomb, J. D. (2001) Biochemistry 40, 2220-2233]. Here, this hypothesis is tested by construction of single and double mutations involving the residues of the Quad mutant. It is shown that mutations of residues that extend into the core structure of MMOB alter many aspects of the MMOH catalyzed reaction but do not mimic the effects of the Quad mutant. In contrast, the MMOB residues that are thought to form part of the interface in the MMOH-MMOB complex increase active site accessibility as observed for the Quad mutant. In particular, the mutant T111A mimics most of the effects of the Quad mutant; thus, Thr111 is proposed to most directly control access. Unexpectedly, mutation of Thr111 to the larger Tyr greatly increases the rate constant for the reaction of larger substrates such as ethane, furan, and nitrobenzene with Q while decreasing the rate constant for the reaction with methane. Other steps in the cycle are dramatically slowed, the regiospecificity for nitrobenzene oxidation is altered, and 10-fold more T111Y than wild-type MMOB is required to maximize the rate of turnover. Thus, T111Y appears to make a more extensive change in local interface structure that allows hydrocarbons at least as large as ethane to bind and react with Q similarly. As a result, the bond cleavage rates for methane, ethane, and their deuterated analogues are shown for the first time to correlate with bond strength in accord with a mechanism in which C-H bond cleavage occurs during reaction of substrates with Q. PMID- 12741819 TI - Asymmetric binding of the high-affinity Q(H)(*)(-) ubisemiquinone in quinol oxidase (bo3) from Escherichia coli studied by multifrequency electron paramagnetic resonance spectroscopy. AB - Ubiquinone-2 (UQ-2) selectively labeled with (13)C (I =(1)/(2)) at either the position 1- or the 4-carbonyl carbon is incorporated into the ubiquinol oxidase bo(3) from Escherichia coli in which the native quinone (UQ-8) has been previously removed. The resulting stabilized anion radical in the high-affinity quinone-binding site (Q(H)(*)(-)) is investigated using multifrequency (9, 34, and 94 GHz) electron paramagnetic resonance (EPR) spectroscopy. The corresponding spectra reveal dramatic differences in (13)C hyperfine couplings indicating a strongly asymmetric spin density distribution over the quinone headgroup. By comparison with previous results on labeled ubisemiquinones in proteins as well as in organic solvents, it is concluded that Q(H)(*)(-) is most probably bound to the protein via a one-sided hydrogen bond or a strongly asymmetric hydrogen bonding network. This observation is discussed with regard to the function of Q(H) in the enzyme and contrasted with the information available on other protein bound semiquinone radicals. PMID- 12741820 TI - Calcium-modulated guanylate cyclase transduction machinery in the photoreceptor- bipolar synaptic region. AB - Rod outer segment membrane guanylate cyclase (ROS-GC) transduction system is a central component of the Ca(2+)-sensitive phototransduction machinery. The system is composed of two parts: Ca(2+) sensor guanylate cyclase activating protein (GCAP) and ROS-GC. GCAP senses Ca(2+) impulses and inhibits the cyclase. This operational feature of the cyclase is considered to be unique and exclusive in the phototransduction machinery. A combination of reconstitution, peptide competition, cross-linking, and immunocytochemical studies has been used in this study to show that the GCAP1/ROS-GC1 transduction system also exists in the photoreceptor synaptic (presynaptic) termini. Thus, the presence of this system and its linkage is not unique to the phototransduction machinery. A recent study has demonstrated that the photoreceptor-bipolar synaptic region also contains a Ca(2+)-stimulated ROS-GC1 transduction system [Duda, T., et al. (2002) EMBO J. 21, 2547-2556]. In this case, S100beta senses Ca(2+) and stimulates the cyclase. The inhibitory and stimulatory Ca(2+)-modulated ROS-GC1 sites are distinct. These findings allow the formation of a new topographic model of ROS-GC1 transduction. In this model, the catalytic module of ROS-GC1 at its opposite ends is flanked by GCAP1 and S100beta modules. GCAP1 senses the Ca(2+) impulse and inhibits the catalytic module; S100beta senses the impulse and stimulates the catalytic module. Thus, ROS-GC1 acts as a bimodal Ca(2+) signal transduction switch in the photoreceptor bipolar synapse. PMID- 12741821 TI - A model for effector activity in a highly specific biological electron transfer complex: the cytochrome P450(cam)-putidaredoxin couple. AB - The camphor hydroxylase cytochrome P450(cam) (CYP101) catalyzes the 5-exo hydroxylation of camphor in the first step of camphor catabolism by Pseudomonas putida. CYP101 forms a specific electron transfer complex with its physiological reductant, the Cys(4)Fe(2)S(2) ferredoxin putidaredoxin (Pdx). Pdx, along with other proteins and small molecules, has also been shown to be an effector for turnover by CYP101. Multidimensional nuclear magnetic resonance (NMR) techniques have been used to make extensive sequential (1)H, (15)N, and (13)C resonance assignments in CYP101 that permit a more complete characterization of the complex formed by CYP101 and Pdx. NMR-detected perturbations in CYP101 upon Pdx binding encompass regions of the CYP101 remote from the putative Pdx binding site, including in particular a region of the CYP101 molecule that has been implicated in substrate access to the active site via dynamical processes. A model for effector activity is proposed in which the primary role of the effector is to prevent uncoupling (formation of reduced oxo species without formation of hydroxycamphor) by enforcing conformations of CYP101 that prevent loss of substrate and/or intermediates prior to turnover. A secondary role could also be to enforce conformations that permit efficient proton transfer into the active site for coupled proton/electron transfer. PMID- 12741822 TI - Zinc-mediated helix capping in a triple-helical protein. AB - Specific sequence signals at alpha-helix termini can assist protein folding by punctuating and cueing secondary structural elements in the final native conformation. Here we report the crystallization of a 56-residue alanine containing peptide, denoted Ala-10(56), in the presence of Zn(2+). The 1.7 A crystal structure shows that Ala-10(56) forms a parallel trimeric coiled coil with three zinc ions anchoring distinct capping conformations at the amino terminal ends of the three helices. In each polypeptide chain, the free alpha amino nitrogen and carbonyl oxygen of the amino-terminal Ser residue coordinate to a Zn(2+) ion to form a five-membered chelate, and the syn-unidentate interaction of the Asp7 side chain with the Zn(2+) cation leads to the formation of a unique docking arrangement for helix capping. Moreover, the coordination of the zinc ion involves a neighboring trimer molecule in the crystal. Consequently, the crystal contacts are stabilized by carboxylate-Zn(2+) interactions between four Ala-10(56) trimers in the crystal lattice. The observed synergy between the protein-zinc ion recognition and the helix-packing arrangements would contribute to the conformational specificity of the Ala-10(56) trimer. PMID- 12741823 TI - New insight into the solution structures of wheat gluten proteins from Raman optical activity. AB - Vibrational Raman optical activity (ROA) spectra of the wheat proteins alpha gliadin (A-gliadin), omega-gliadin, and a 30 kDa peptide called T-A-1 from the high molecular weight glutenin subunit (HMW-GS) Dx5 were measured to obtain new information about their solution structures. The spectral data show that, under the conditions investigated, A-gliadin contains a considerable amount of hydrated alpha-helix, most of which probably lies within a relatively structured C terminal domain. Smaller quantities of beta-structure and poly(l-proline) II (PPII) helix were also identified. Addition of methanol was found to increase the alpha-helix content at the expense of some of the beta and PPII structure. In comparison, omega-gliadin and the T-A-1 peptide were found to consist of large amounts of well-defined PPII structure with some turns but no alpha-helix. The results for the T-A-1 peptide are in agreement with a model in which HMW-GS are extended but not highly rigid. Application of a pattern recognition technique, based on principal component analysis (PCA), to the ROA spectra reinforces these conclusions. PMID- 12741824 TI - In vitro evolution of the binding specificity of neocarzinostatin, an enediyne binding chromoprotein. AB - Neocarzinostatin is the most studied member of the enediyne-chromoprotein family, and is clinically used as an antitumoral agent. Neocarzinostatin could be a promising drug delivery vehicle if new binding specificities could be conferred to its protein scaffold. We used in vitro evolution methods to demonstrate that this approach is feasible. We created large libraries containing between 1.7 x 10(8) and 1.4 x 10(9) independent clones, where up to 13 side chains pointing toward the binding crevice were randomly substituted. We then used phage display to select variants that bind to a model ligand (testosterone) which is unrelated to the natural ligand of neocarzinostatin. Several different binders were selected from each library. The corresponding proteins were expressed in Escherichia coli and their affinities and specificities were characterized in detail. K(D) values of about 20 nM were obtained for streptavidin-bound testosterone. The K(D) of selected proteins for free soluble testosterone are between 7 and 55 microM and therefore higher than the K(D) for streptavidin-bound testosterone. The spacer and streptavidin used during selection contributed to the high affinity of the selected binders for the target. Binding studies of 15 different steroids related to testosterone allowed us to determine that C3, 4, 5, 6, and 7 on cycles A and B and the conjugated 3 oxo group of the steroid molecule were essential for molecular recognition. Other testosterone analogues substituted on C1, 2, 9, 11, 15, and 17 were not discriminated from testosterone. These results demonstrate that the binding specificity of this protein family can be extended to compounds that are completely unrelated to the natural enediyne chromophore family. This type of highly expressed, stable proteins with tailored binding properties have a wide potential range of applications. PMID- 12741825 TI - Structure and mechanism of Pseudomonas aeruginosa PhzD, an isochorismatase from the phenazine biosynthetic pathway. AB - PhzD from Pseudomonas aeruginosa is an isochorismatase involved in phenazine biosynthesis. Phenazines are antimicrobial compounds that provide Pseudomonas with a competitive advantage in certain environments and may be partly responsible for the persistence of Pseudomonas infections. In vivo, PhzD catalyzes the hydrolysis of the vinyl ether functional group of 2-amino-2 deoxyisochorismate, yielding pyruvate and trans-2,3-dihydro-3-hydroxyanthranilic acid, which is then utilized in the phenazine biosynthetic pathway. PhzD also catalyzes hydrolysis of the related vinyl ethers isochorismate, chorismate, and 4 amino-4-deoxychorismate. Here we report the 1.5 A crystal structure of native PhzD, and the 1.6 A structure of the inactive D38A variant in complex with isochorismate. The structures reveal that isochorismate binds to the PhzD active site in a trans-diaxial conformation, and superposition of the structures indicates that the methylene pyruvyl carbon of isochorismate is adjacent to the side chain carboxylate of aspartate 38. The proximity of aspartate 38 to isochorismate and the complete loss of activity resulting from the conversion of aspartate 38 to alanine suggest a mechanism in which the carboxylate acts as a general acid to protonate the substrate, yielding a carbocation/oxocarbonium ion that is then rapidly hydrated to form a hemiketal intermediate, which then decomposes spontaneously to products. The structure of PhzD is remarkably similar to other structures from a subfamily of alpha/beta-hydrolase enzymes that includes pyrazinamidase and N-carbamoylsarcosine amidohydrolase. However, PhzD catalyzes unrelated chemistry and lacks a nucleophilic cysteine found in its close structural relatives. The vinyl ether hydrolysis catalyzed by PhzD represents yet another example of the catalytic diversity seen in the alpha/beta hydrolase family, whose members are also known to hydrolyze amides, phosphates, phosphonates, epoxides, and C-X bonds. PMID- 12741826 TI - Ionic states of substrates and transition state analogues at the catalytic sites of N-ribosyltransferases. AB - Purine nucleoside phosphorylase (PNP) and hypoxanthine-guanine phosphoribosyltransferase (HGPRTase) catalyze N-ribosidic bond cleavage in purine nucleosides and nucleotides, with addition of phosphate or pyrophosphate to form phosphorylated alpha-D-ribose products. The transition states have oxacarbenium ion character with a positive charge near 1'-C and ionic stabilization from nearby phosphoryl anions. Immucillin-H (ImmH) and Immucillin-H 5'-PO(4) (ImmHP) resemble the transition state charge when protonated at 4'-N and bind tightly to these enzymes with K(d) values of 20 pM to 1 nM. It has been proposed that Immucillins bind as the 4'-N neutral form and are protonated in the slow-onset step. Solution and solid-state NMR spectra of ImmH, ImmHP, guanosine, and GMP in complexes with two PNPs and a HGPRTase have been used to characterize their ionization states. Results with PNP*ImmH*PO(4) and HGPRTase*ImmHP*MgPP(i) indicate protonation at N-4' for the tightly bound inhibitors. The 1'-(13)C and 1'-(1)H resonances of bound Immucillins showed large downfield shifts as compared to Michaelis complexes, suggesting distortion of 1'-C toward sp(2) geometry. The Immucillins act as transition state mimics by binding with neutral iminoribitol groups followed by 4'-N protonation during slow-onset inhibition to form carbocationic mimics of the transition states. The ability of the Immucillins to mimic both substrate and transition state features contributes to their capture of transition state binding energy. Enzyme-activated phosphoryl nucleophiles bound to PNP and HGPRTase suggest enhanced electrostatic stabilization of the cationic transition states. Distortion of the oxacarbenium ion mimic toward transition state geometry is a common feature of the three distinct enzymatic complexes analyzed here. Substrate complexes, even in catalytically cycling equilibrium mixtures, do not reveal similar distortions. PMID- 12741827 TI - Structural basis for feedback inhibition of the deoxyribonucleoside salvage pathway: studies of the Drosophila deoxyribonucleoside kinase. AB - Deoxyribonucleoside kinases are feedback inhibited by the final products of the salvage pathway, the deoxyribonucleoside triphosphates. In the present study, the mechanism of feedback inhibition is presented based on the crystal structure of a complex between the fruit fly deoxyribonucleoside kinase and its feedback inhibitor deoxythymidine triphosphate. The inhibitor was found to be bound as a bisubstrate inhibitor with its nucleoside part in the nucleoside binding site and with its phosphate groups partially occupying the phosphate donor site. The overall structure of the enzyme--inhibitor complex is very similar to the enzyme- substrate complexes with deoxythymidine and deoxycytidine, except for a conformational change within a region otherwise directly involved in catalysis. This conformational change involves a magnesium ion, which is coordinated in the inhibitor complex to the phosphates and to the primary base, Glu52, that normally is positioned close to the 5'-OH of the substrate deoxyribose. PMID- 12741828 TI - Quaternary structure of aldolase leads to differences in its folding and unfolding intermediates. AB - Pulsed hydrogen exchange mass spectrometry has been used to investigate folding of rabbit muscle aldolase, an alpha/beta-barrel protein exhibiting the classic TIM structure. Aldolase unfolded in GdHCl refolded as the denaturant concentration was reduced by dialysis. Samples withdrawn during dialysis were pulse-labeled with deuterium to identify unfolded regions in structural forms highly populated during the folding process. Intact, labeled aldolase was digested into fragments, which were analyzed by HPLC electrospray ionization mass spectrometry to detect the H/D exchange along the aldolase backbone. For some concentrations of GdHCl, bimodal distributions of deuterium were found for most peptic fragments, indicating that regions represented by these fragments were either unfolded or folded in the intact polypeptide prior to labeling. The extent of folding was determined from these mass spectra, as well as by CD (220 nm) and enzymatic activity. These results show that folding to the active form involves three domains and two intermediates. Approximately 110 residues fold to highly compact forms in each step. These results also show that each folding domain includes widely separated regions of the backbone. When compared with the results of a previous study of aldolase unfolding, these results show that the folding and unfolding domains include most of the same residues. However, three short segments change domains depending on whether the process is folding or unfolding. These changes are attributed to the very stable quaternary structure of rabbit muscle aldolase. PMID- 12741829 TI - Proline isomerization in bovine pancreatic ribonuclease A. 2. Folding conditions. AB - The kinetics of cis-trans isomerization of individual X-Pro peptide groups is used to study the backbone dynamics of bovine pancreatic ribonuclease A (RNase A). We previously developed and validated a fluorescence method for monitoring the cis-trans isomerization of the Tyr92-Pro93 and Asn113-Pro114 peptide groups of RNase A under unfolding conditions [Juminaga, D., Wedemeyer, W. J., and Scheraga, H. A. (1998) Biochemistry 37, 11614-11620]. The essence of this method is to introduce a fluorescent residue (Tyr or Trp) in a position adjacent to the isomerizing proline (if one is not already present) and to eliminate the fluorescence of other such residues adjacent to prolines by mutating them to phenylalanine. Here, we extend this method to observe the cis-trans isomerization of these peptide groups under folding conditions using two site-directed mutants (Y92F and Y115F) of RNase A. Both isomerizations decelerate with increasing concentrations of GdnHCl, with nearly identical m values (1.11 and 1.19 M(-1), respectively) and extrapolated zero-GdnHCl time constants (42 and 32 s, respectively); by contrast, under unfolding conditions, the cis-trans isomerizations of both Pro93 and Pro114 are independent of GdnHCl concentration. Remarkably, the isomerization rates under folding conditions at GdnHCl concentrations above 1 M are significantly slower than those measured under unfolding conditions. The temperature dependence of the Pro114 isomerization under folding conditions is also unusual; whereas Pro93 exhibits an activation energy typical of proline isomerization (19.4 kcal/mol), Pro114 exhibits a sharply reduced activation energy of 5.7 kcal/mol. A structurally plausible model accounts for these results and, in particular, shows that folding conditions strongly accelerate the cis-trans isomerization of both peptide groups to their native cis conformation, suggesting the presence of flickering local structure in their beta-hairpins. PMID- 12741830 TI - Hydroperoxidase II of Escherichia coli exhibits enhanced resistance to proteolytic cleavage compared to other catalases. AB - Catalase (hydroperoxidase) HPII of Escherichia coli is the largest catalase so far characterized, existing as a homotetramer of 84 kDa subunits. Each subunit has a core structure that closely resembles small subunit catalases, supplemented with an extended N-terminal sequence and compact flavodoxin-like C-terminal domain. Treatment of HPII with trypsin, chymotrypsin, or proteinase K, under conditions of limited digestion, resulted in cleavage of 72-74 residues from the N-terminus of each subunit that created a homotetramer of 76 kDa subunits with 80% of wild-type activity. Longer treatment with proteinase K removed the C terminal domain, producing a transient 59 kDa subunit which was subsequently cleaved into two fragments, 26 and 32 kDa. The tetrameric structure was retained despite this fragmentation, with four intermediates being observed between the 336 kDa native form and the 236 kDa fully truncated form corresponding to tetramers with a decreasing complement of C-termini (4, 3, 2, and 1). The truncated tetramers retained 80% of wild-type activity. The T(m) for loss of activity during heating was decreased from 85 to 77 degrees C by removal of the N terminal sequence and to 59 degrees C by removal of the C-terminal domain, revealing the importance of the C-terminal domain in enzyme stability. The sites of cleavage were determined by N- and C-terminal sequencing, and two were located on the surface of the tetramer with a third being exposed by removal of the C terminal domain. PMID- 12741831 TI - The Euplotes La motif protein p43 has properties of a telomerase-specific subunit. AB - Telomerase is a specialized reverse transcriptase synthesizing DNA repeats at telomeres. In addition to the RNA and catalytic protein components, telomerase from the ciliate Euplotes aediculatus contains the subunit p43. This protein is homologous to the La autoantigen, functioning in maturation of RNA polymerase III transcripts. Here we provide evidence that p43 is primarily associated with the telomerase ribonucleoprotein in vivo. Recombinant p43 binds telomerase RNA with low-nanomolar affinity in vitro, recognizing stem I and adjacent nucleotides or structures in the core of the RNA. Unlike authentic La proteins, p43 does not bind strongly to RNA polymerase III precursor transcripts and does not exhibit a marked binding preference for 3'-terminal oligouridylate residues. In isolated macronuclei, p43 largely colocalizes with telomerase RNA in discrete foci. These findings suggest that p43 is not the Euplotes La protein but instead plays a dedicated role in telomerase assembly and/or function. Thus, p43 joins the telomerase reverse transcriptase and the yeast proteins Est1p and Est3p as the only telomerase-specific proteins identified so far. PMID- 12741832 TI - Structure-function analysis of the auxilin J-domain reveals an extended Hsc70 interaction interface. AB - J-domains are widespread protein interaction modules involved in recruiting and stimulating the activity of Hsp70 family chaperones. We have determined the crystal structure of the J-domain of auxilin, a protein which is involved in uncoating clathrin-coated vesicles. Comparison to the known structures of J domains from four other proteins reveals that the auxilin J-domain is the most divergent of all J-domain structures described to date. In addition to the canonical J-domain features described previously, the auxilin J-domain contains an extra N-terminal helix and a long loop inserted between helices I and II. The latter loop extends the positively charged surface which forms the Hsc70 binding site, and is shown by directed mutagenesis and surface plasmon resonance to contain side chains important for binding to Hsc70. PMID- 12741833 TI - Isoform specific differences in binding of a dual-specificity A-kinase anchoring protein to type I and type II regulatory subunits of PKA. AB - Dual-specificity AKAPs bind to type I (RI) and type II (RII) regulatory subunits of cAMP-dependent protein kinase A (PKA), potentially recruiting distinct cAMP responsive holoenzymes to a given intracellular location. To understand the molecular basis for this "dual" functionality, we have examined the pH dependence, the salt-dependence, and the kinetics of binding of the A-kinase binding (AKB) domain of D-AKAP2 to the regulatory subunit isoforms of PKA. Using fluorescence anisotropy, we have found that a 27-residue peptide corresponding to the AKB domain of D-AKAP2 bound 25-fold more tightly to RIIalpha than to RIalpha. The higher affinity for RIIalpha was the result of a slower off-rate as determined by surface plasmon resonance. The high-affinity interaction for RIalpha and RIIalpha was pH-independent from pH 7.4 to 5.0. At pH 4.0, both isoforms had a reduction in binding affinity. Additionally, binding of the AKB domain to RIalpha was independent of solution ionic strength, whereas RIIalpha had an increased binding affinity at higher ionic strength. This suggests that the relative energetic contribution of the charge stabilization is different for the two isoforms. This prediction was confirmed by mutagenesis in which acidic mutations, primarily of E10 and D23, in the AKB domain affected binding to RIalpha but not to RIIalpha. These isoform-specific differences provide a foundation for developing isoform-specific peptide inhibitors of PKA anchoring by dual-specificity AKAPs, which can be used to evaluate the physiological significance of dual-specificity modes of PKA anchoring. PMID- 12741835 TI - A side reaction of alanine racemase: transamination of cycloserine. AB - Alanine racemase (EC 5.1.1.1) catalyzes the interconversion of alanine enantiomers, and thus represents the first committed step involved in bacterial cell wall biosynthesis. Cycloserine acts as a suicide inhibitor of alanine racemase and as such, serves as an antimicrobial agent. The chemical means by which cycloserine inhibits alanine racemase is unknown. Through spectroscopic assays, we show here evidence of a pyridoxal derivative (arising from either isomer of cycloserine) saturated at the C4' carbon position. We additionally report the L- and D-cycloserine inactivated crystal structures of Bacillus stearothermophilus alanine racemase, which corroborates the spectroscopy via evidence of a 3-hydroxyisoxazole pyridoxamine derivative. Upon the basis of the kinetic and structural properties of both the L- and D-isomers of the inhibitor, we propose a mechanism of alanine racemase inactivation by cycloserine. This pathway involves an initial transamination step followed by tautomerization to form a stable aromatic adduct, a scheme similar to that seen in cycloserine inactivation of aminotransferases. PMID- 12741834 TI - Reactions of Mycobacterium tuberculosis truncated hemoglobin O with ligands reveal a novel ligand-inclusive hydrogen bond network. AB - Truncated hemoglobin O (trHbO) is one of two trHbs in Mycobacterium tuberculosis. Remarkably, trHbO possesses two novel distal residues, in addition to the B10 tyrosine, that may be important in ligand binding. These are the CD1 tyrosine and G8 tryptophan. Here we investigate the reactions of trHbO and mutants using stopped-flow spectrometry, flash photolysis, and UV-enhanced resonance Raman spectroscopy. A biphasic kinetic behavior is observed for combination and dissociation of O(2) and CO that is controlled by the B10 and CD1 residues. The rate constants for combination (<1.0 microM(-1) s(-1)) and dissociation (<0.006 s(-1)) of O(2) are among the slowest known, precluding transport or diffusion of O(2) as a major function. Mutation of CD1 tyrosine to phenylalanine shows that this group controls ligand binding, as evidenced by 25- and 77-fold increases in the combination rate constants for O(2) and CO, respectively. In support of a functional role for G8 tryptophan, UV resonance Raman indicates that the chi((2,1)) dihedral angle for the indole ring increases progressively from approximately 93 degrees to at least 100 degrees in going sequentially from the deoxy to CO to O(2) derivative, demonstrating a significant conformational change in the G8 tryptophan with ligation. Remarkably, protein modeling predicts a network of hydrogen bonds between B10 tyrosine, CD1 tyrosine, and G8 tryptophan, with the latter residues being within hydrogen bonding distance of the heme-bound ligand. Such a rigid hydrogen bonding network may thus represent a considerable barrier to ligand entrance and escape. In accord with this model, we found that changing CD1 or B10 tyrosine for phenylalanine causes only small changes in the rate of O(2) dissociation, suggesting that more than one hydrogen bond must be broken at a time to promote ligand escape. Furthermore, trHbO-CO cannot be photodissociated under conditions where the CO derivative of myoglobin is extensively photodissociated, indicating that CO is constrained near the heme by the hydrogen bonding network. PMID- 12741836 TI - Iron-sulfur cluster biosynthesis. A comparative kinetic analysis of native and Cys-substituted ISA-mediated [2Fe-2S]2+ cluster transfer to an apoferredoxin target. AB - ISA type proteins mediate cluster transfer to apoprotein targets. Rate constants have been determined for cluster transfer from Schizosaccharomyces pombe ISA to apo Fd. Substitution of the cysteine residues of ISA produced derivative proteins (C72A, C136A, and C138A) that were found to be at least as active in cluster transfer reactions as the native form at 25 degrees C (k(2) approximately 170 M( 1) min(-1) for native, k(2) approximately 169 M(-1) min(-1) for C72A, k(2) approximately 206 M(-1) min(-1) for C136A, and k(2) approximately 242 M(-1) min( 1) for C138A), although the yield of cluster transfer was found to be lower as a consequence of the enhanced lability of clusters in the derivative proteins. Minor variations in rate constant for the ISA Cys derivatives do not reflect any change in the affinity of binding to the apo Fd since k(2) was found to be independent of the concentration of apo Fd over the range of 1-25 microM. The pH dependence of cluster transfer rates was found to be similar for native and C136A ISA, with an observed pK(a) of 7.8 determined from the pH profiles for cluster transfer activity of each protein. The temperature dependence of the rate constant defining the cluster transfer reaction for the wild type versus this C136A ISA derivative is distinct (DeltaH* approximately 6.3 kcal mol(-1) and DeltaS* approximately -27.3 cal K(-1) mol(-1) for native and DeltaH* approximately 2.7 kcal mol(-1) and DeltaS* approximately -38.9 cal K(-1) mol(-1) for C136A ISA). Instability of the protein-bound cluster precluded a comparison with data from pH and temperature dependencies for the two other Cys derivatives. Experiments to determine the dependence of reaction rate constants on viscosity indicate cluster transfer is rate-limiting. A comparison of cross-species rate constants for cluster transfer to apo Fd targets from Homo sapiens and S. pombe demonstrated that the identity of the Fd is less critical for promoting cluster transfer from Sp ISA (at 25 degrees C, k(2) approximately 170 M(-1) min(-1) for Sp Fd and k(2) approximately 169 M(-1) min(-1) for Hs Fd). This contrasts with an earlier observation for ISU-mediated cluster assembly [Wu, S., et al. (2002) Biochemistry 41, 8876-8885], where the rates differed for Hs and Sp target Fd's, suggesting distinct binding sites for binding of holo ISA and ISU to apo Fd. PMID- 12741838 TI - Enrichment of the light-harvesting complex in diadinoxanthin and implications for the nonphotochemical fluorescence quenching in diatoms. AB - The pigment composition of diatoms differs from that of green algae and plants. Diatoms contain chlorophyll (Chl(1)) c, fucoxanthin, and diadinoxanthin (DD). An intermittent light regime during growth induced a large increase in the DD content in the marine planktonic diatom Phaeodactylum tricornutum. Light harvesting complex containing fucoxanthin (LHCF) subunits were purified on a sucrose gradient after treatment of thylakoid membranes with a mild detergent. DD was found in all the LHCF fractions: a "major" composite LHCF fraction and the two fractions where some LHCF was associated with photosystem centers. For cells enriched in DD, most of the additional DD molecules were bound to the major LHCF fraction. The DD enrichment of the major LHCF fraction was accompanied by a decrease in the fucoxanthin to Chl a ratio. Either some fucoxanthin molecules were replaced by DD or there could be a relative enrichment of subunits rich in DD at the expense of fucoxanthin/Chl c rich subunits. Under high light illumination, a higher degree of de-epoxidation of DD into DT was observed for the major LHCF of cells enriched in DD. This fraction has the higher DD content and the higher degree of de-epoxidation. These results show that the distal antennae, probably mostly isolated as the major LHCF fraction, play a crucial role in the formation of NPQ, its amplitude depending on the amount of DD bound and on the degree of de-epoxidation (Lavaud et al. (2002) Plant Physiol. 129, 1398-1406). PMID- 12741837 TI - Interaction with membrane lipids and heme ligand binding properties of Escherichia coli flavohemoglobin. AB - Escherichia coli flavohemoglobin has been shown to be able to bind specifically unsaturated and/or cyclopropanated fatty acids with very high affinity. Unsaturated or cyclopropanated fatty acid binding results in a modification of the visible absorption spectrum of the ferric heme, corresponding to a transition from a pentacoordinated (typical of the ligand free protein) to a hexacoordinated, high spin, heme iron. In contrast, no detectable interaction has been observed with saturated fatty acid, saturated phospholipids, linear, cyclic, and aromatic hydrocarbons pointing out that the protein recognizes specifically double bonds in cis conformation within the hydrocarbon chain of the fatty acid molecule. Accordingly, as demonstrated in gel filtration experiments, flavohemoglobin is able to bind liposomes obtained from lipid extracts of E. coli membranes and eventually abstract phospholipids containing cis double bonds and/or cyclopropane rings along the acyl chains. The presence of a protein bound lipid strongly affects the thermodynamic and kinetic properties of imidazole binding to the ferric protein and brings about significant modifications in the reactivity of the ferrous protein with oxygen and carbon monoxide. The effect of the bound lipid has been accounted for by a reaction scheme that involves the presence of two sites for the lipid/ligand recognition, namely, the heme iron and a non-heme site located in a loop region above the heme pocket. PMID- 12741839 TI - All-trans-retinyl esters are the substrates for isomerization in the vertebrate visual cycle. AB - The identification of the critical enzyme(s) that carries out the trans to cis isomerization producing 11-cis-retinol during the operation of the visual cycle remains elusive. Confusion exists in the literature as to the exact nature of the isomerization substrate. At issue is whether it is an all-trans-retinyl ester or all-trans-retinol (vitamin A). As both putative substrates interconvert rapidly in retinal pigment epithelial membranes, the choice of substrate can be ambiguous. The two enzymes that effect interconversion of all-trans-retinol and all-trans-retinyl esters are lecithin retinol acyl transferase (LRAT) and retinyl ester hydrolase (REH). The retinyl ester or all-trans-retinol pools are radioactively labeled separately in the presence of inhibitors of LRAT and REH, effectively preventing their interconversion. Pulse-chase experiments unambiguously demonstrate that all-trans-retinyl esters, and not all-trans retinol, are the precursors of 11-cis-retinol. When the all-trans-retinyl ester pool is radioactively labeled, the resulting 11-cis-retinol is labeled with the same specific activity as the precursor ester. The converse is true with vitamin A. These data unambiguously establish all-trans-retinyl esters as the precursors of 11-cis-retinol. PMID- 12741840 TI - Quinone (Q(B)) binding site and protein stuctural changes in photosynthetic reaction center mutants at Pro-L209 revealed by vibrational spectroscopy. AB - The effect of substituting Pro-L209 with Tyr, Phe, Glu, and Thr in photosynthetic reaction centers (RCs) from Rhodobacter sphaeroides was investigated by monitoring the light-induced FTIR absorption changes associated with the photoreduction of the secondary quinone Q(B). Pro-L209 is close to a chain of ordered water molecules connecting Q(B) to the bulk phase. In wild-type RCs, two distinct main Q(B) binding sites (distal and proximal to the non-heme iron) have been described in the literature. The X-ray structures of the mutant RCs Pro-L209 --> Tyr, Pro-L209 --> Phe, and Pro-L209 --> Glu have revealed that Q(B) occupies a proximal, intermediate, and distal position, respectively [Kuglstatter, A., Ermler, U., Michel, H., Baciou, L., and Fritzsch, G. (2001) Biochemistry 40, 4253 4260]. FTIR absorption changes associated with the reduction of Q(B) in Pro-L209 -> Phe RCs reconstituted with (13)C-labeled ubiquinone show a highly specific IR fingerprint for the C=O and C=C modes of Q(B) upon selective labeling at C(1) or C(4). This IR fingerprint is similar to those of wild-type RCs and the Pro-L209 - > Tyr mutant [Breton, J., Boullais, C., Mioskowski, C., Sebban, P., Baciou, L., and Nabedryk, E. (2002) Biochemistry 41, 12921-12927], demonstrating that equivalent interactions occur between neutral Q(B) and the protein in wild-type and mutant RCs. It is concluded that in all RCs, neutral Q(B) in its functional state occupies a unique binding site which is favored to be the proximal site. This result contrasts with the multiple Q(B) binding sites found in crystal structures. With respect to wild-type RCs, the largest FTIR spectral changes upon Q(B)(-) formation are observed for the Phe-L209 and Tyr-L209 mutants which undergo similar protein structural changes and perturbations of the semiquinone modes. Smaller changes are observed for the Glu-L209 mutant, while the vibrational properties of the Thr-L209 mutant are essentially the same as those for native RCs. PMID- 12741841 TI - Ascorbate-mediated LHCII protein phosphorylation--LHCII kinase regulation in light and in darkness. AB - A freeze-thaw cycle of isolated thylakoids in darkness in the presence of ascorbate was employed as a novel experimental system to activate the light harvesting complex (LHC) II kinase. Under these conditions ascorbate reduces Q(A), the primary quinone electron acceptor of photosystem (PS) II, and the subsequent reduction of plastoquinone and the cytochrome (cyt) b(6)f complex results in the activation of the LHCII kinase. Using this activation system, several facets of regulation of LHCII protein phosphorylation were unravelled. (i) Myxothiazol inhibited the activation of LHCII protein phosphorylation, thus being a potent inhibitor of electron flow not only in cyt bc complexes but in darkness also in cyt b(6)f complexes. (ii) Oxygen, the only electron acceptor in darkness, was required for LHCII kinase activation demonstrating that after a full reduction of the cyt b(6)f complex, an additional plastoquinol oxidation cycle in the quinol oxidation (Qo) site is required for LHCII kinase activation. (iii) In the absence of electron flow, when the intersystem electron carriers are reduced, the activated LHCII kinase has a half-life of 40 min, whereas the fully activated LHCII kinase becomes deactivated in a time scale of seconds upon oxidation of the cyt b(6)f complex, indicating that the kinase constantly reads the redox poise of the cyt b(6)f complex. (iv) The LHCII kinase is more tightly bound to the thylakoid membrane than the PS II core protein kinase(s). It is concluded that oxidation of plastoquinol at the Qo site of the reduced cyt b(6)f complex is required for LHCII kinase activation, while rapid reoccupation of the Qo site with plastoquinol is crucial for sustenance of the active state of the LHCII kinase. PMID- 12741842 TI - Hydrogen exchange kinetics of RNase A and the urea:TMAO paradigm. AB - A key paradigm in the biology of adaptation holds that urea affects protein function by increasing the fluctuations of the native state, while trimethylamine N-oxide (TMAO) affects function in the opposite direction by decreasing the normal fluctuations of the native ensemble. Using urea and TMAO separately and together, hydrogen exchange (HX) studies on RNase A at pH* 6.35 were used to investigate the basic tenets of the urea:TMAO paradigm. TMAO (1 M) alone decreases HX rate constants of a select number of sites exchanging from the native ensemble, and low urea alone increases the rate constants of some of the same sites. Addition of TMAO to urea solutions containing RNase A also suppresses HX rate constants. The data show that urea and TMAO independently or in combination affect the dynamics of the native ensemble in opposing ways. The results provide evidence in support of the counteraction aspect of the urea:TMAO paradigm linking structural dynamics with protein function in urea-rich organs and organisms. RNase A is so resistant to urea denaturation at pH* 6.35 that even in the presence of 4.8 M urea, the native ensemble accounts for >99.5% of the protein. An essential test, devised to determine the HX mechanism of exchangeable protons, shows that over the 0-4.8 M urea concentration range nearly 80% of all observed sites convert from EX2 to EX1. The slow exchange sites are all EX1; they do not exhibit global exchange even at urea concentrations (5.8 M) well into the denaturation transition zone, and their energetically distinct activated complexes leading to exchange gives evidence of residual structure. Under these experimental conditions, the use of DeltaG(HX) as a basis for HX analysis of RNase A urea denaturation is invalid. PMID- 12741843 TI - Reductive half-reaction of nitroalkane oxidase: effect of mutation of the active site aspartate to glutamate. AB - The flavoenzyme nitroalkane oxidase catalyzes the oxidation of primary and secondary nitroalkanes to the respective aldehydes or ketones, releasing nitrite. The enzyme has recently been identified as being homologous to the acyl-CoA dehydrogenase family of enzymes [Daubner, S. C., Gadda, G., Valley, M. P., and Fitzpatrick, P. F. (2002) Proc. Natl. Acad. Sci. U.S.A. 99, 2702-2707]. The glutamate which acts as an active site base in that family of enzymes aligns with Asp402 of nitroalkane oxidase. To evaluate the identification of Asp402 as an active site base, the effect of mutation of Asp402 to glutamate on the rate of cleavage of the nitroalkane C-H bond has been determined. Deuterium kinetic isotope effects on steady state kinetic parameters and direct measurement of the rate of flavin reduction establish that the mutation increases the DeltaG(++) for C-H bond cleavage by 1.6-1.9 kcal/mol. There is no effect on the rate of reaction of the reduced enzyme with oxygen. These results support the assignment of Asp402 as the active site base in nitroalkane oxidase. PMID- 12741844 TI - Rapid-mix and chemical quench studies of ferredoxin-reduced stearoyl-acyl carrier protein desaturase. AB - Stearoyl-ACP Delta9 desaturase (Delta9D) catalyzes the NADPH- and O(2)-dependent insertion of a cis double bond between the C9 and C10 positions of stearoyl-ACP (18:0-ACP) to produce oleoyl-ACP (18:1-ACP). This work revealed the ability of reduced [2Fe-2S] ferredoxin (Fd) to act as a catalytically competent electron donor during the rapid conversion of 18:0-ACP into 18:1-ACP. Experiments on the order of addition for substrate and reduced Fd showed high conversion of 18:0-ACP to 18:1-ACP (approximately 95% per Delta9D active site in a single turnover) when 18:0-ACP was added prior to reduced Fd. Reactions of the prereduced enzyme substrate complex with O(2) and the oxidized enzyme-substrate complex with reduced Fd were studied by rapid-mix and chemical quench methods. For reaction of the prereduced enzyme-substrate complex, an exponential burst phase (k(burst) = 95 s(-1)) of product formation accounted for approximately 90% of the turnover expected for one subunit in the dimeric protein. This rapid phase was followed by a slower phase (k(linear) = 4.0 s(-1)) of product formation corresponding to the turnover expected from the second subunit. For reaction of the oxidized enzyme substrate complex with excess reduced Fd, a slower, linear rate (k(obsd) = 3.4 s( 1)) of product formation was observed over approximately 1.5 turnovers per Delta9D active site potentially corresponding to a third phase of reaction. An analysis of the deuterium isotope effect on the two rapid-mix reaction sequences revealed only a modest effect on k(burst) ((D)k(burst) approximately 1.5) and k(linear) (D)k(linear) approximately 1.4), indicating C-H bond cleavage does not contribute significantly to the rate-limiting steps of pre-steady-state catalysis. These results were used to assemble and evaluate a minimal kinetic model for Delta9D catalysis. PMID- 12741845 TI - Importance of the D and E helices of the molecular chaperone DnaK for ATP binding and substrate release. AB - The C-terminal domain of the molecular chaperone DnaK is a compact lid-like structure made up of five alpha-helices (alphaA-alphaE) (residues 508-608) that is followed by a 30-residue disordered, flexible region (609-638). The lid encapsulates the peptide molecule bound in the substrate-binding domain, whereas the function of the 30-residue disordered region is not known. By sequentially deleting the flexible subdomain and the individual lid helices, we deduced the importance of each structural unit to creating long-lived DnaK-peptide complexes. Here we report that (i) the alphaD helix is essential for long-lived DnaK-peptide complexes. For example, ATP triggers the dissociation of a acrylodan-labeled p5 peptide (ap5, a-CLLLSAPRR) from wtDnaK and DnaK595(A-D) with k(off) equal to 7.6 and 8.9 s(-1), respectively, whereas when the D-helix is deleted, creating DnaK578(A-C), k(off) jumps to 207 s(-1). (ii) The presence of the alphaB helix impacts the rate of the ATP-induced high-to-low affinity conformational change. For example, ATP induces this conformational change in a lidless variant, DnaK517(1/2A), with a rate constant of 442 s(-1), whereas, after adding back the B-helix (residues 518-554), ATP induces this conformational change in DnaK554(A B) with a rate constant of 2.5 s(-1). Our interpretation is that this large decrease occurs because the B-helix of the DnaK554(A-B) is bound in the substrate binding site. (iii) The deletion analysis also revealed that residues 596-638, which comprise the alphaE helix and the flexible subdomain, affect ATP binding. Our results are consistent with this part of the lid producing conformational heterogeneity, perhaps by binding to the ATPase domain. PMID- 12741846 TI - Energy transduction optical sensor in skeletal myosin. AB - The skeletal myosin cross-bridge in dynamic association with actin is the unitary energy transducer in muscle, converting free energy from ATP hydrolysis into contractile force. Myosin's conserved ATP-sensitive tryptophan (AST) is an energy transduction optical sensor signaling transduction-related transient conformation change by modulating its fluorescence intensity amplitude and relaxation rate. Recently introduced techniques have provided the means of observing the time resolved intensity decay from this single residue in the native protein to elucidate the mechanism of its ATP sensitivity. AST signal characteristics could be derived from local protein structure by a scenario involving interactions with excited-state tryptophan. This investigation suggests the very different possibility that hypochromism induced in the tryptophan absorption band, a ground state effect, is a significant structural effector of optical transduction sensing. This possibility makes feasible the interpretation of the transient AST optical signal in terms of dynamical protein structure, thereby raising the empirical signal to the level of a structural determinant. Using the crystallographically based geometry from several myosin structures, the maximum calculated AST hypochromism is <10% to be compared with the value of approximately 30% observed here experimentally. Rationalizing the discrepancy invites further investigation of S1 dynamical structure local to the AST during transduction. PMID- 12741847 TI - Dimerization and inter-chromophore distance of Cph1 phytochrome from Synechocystis, as monitored by fluorescence homo and hetero energy transfer. AB - We investigated the dimerization of phytochrome Cph1 from the cyanobacterium Synechocystis by fluorescence resonance energy transfer (FRET). As donor we used the chromophore analogue phycoerythrobilin (PEB) and as acceptor either the natural chromophore phycocyanobilin (PCB; hetero transfer) or PEB (homo transfer). Both chromophores bind in a 1:1 stoichiometry to apo-monomers expressed in Escherichia coli. Energy transfer was characterized by time-resolved fluorescence intensity and anisotropy decay after excitation of PEB by picosecond pulses from a tunable Ti-sapphire laser system. ApoCph1 was first assembled with PEB at a low stoichiometry of 0.1. The remaining sites were then sequentially titrated with PCB. In the course of this titration, the mean lifetime of PEB decreased from 3.33 to 1.25 ns in the P(r) form of Cph1, whereas the anisotropy decay was unaffected. In the P(fr)/P(r) photoequilibrium (about 65% P(fr)), the mean lifetime decreased significantly less, to 1.67 ns. These observations provide strong support for inter-chromophore hetero energy transfer in mixed PEB/PCB dimers. The reduced energy transfer in P(fr) may be due to a structural difference but is at least in part due to the difference in spectral overlap, which was 4.1 x 10(-13) and 1.6 x 10(-13) cm(3) M(-1) in P(r) and P(fr), respectively. From the changes in the mean lifetime, rates of hetero energy transfer of 0.68 and 0.37 ns(-1) were calculated for the P(r) form and the P(fr)/P(r) photoequilibrium, respectively. Sequential titration of apo Cph1 with PEB alone to full occupancy did not affect the intensity decay but led to a substantial increase in depolarization. This is the experimental signature of homo energy transfer. Values for the rate of energy transfer k(HT) (0.47 ns(-1)) and the angle 2theta between the transition dipole moment directions (2theta = 45 +/- 5 degrees) were determined from an analysis of the concentration dependence of the anisotropy at five different PEB/Cph1 stoichiometries. The independently determined rates of hetero and homo energy transfer are thus of comparable magnitude and consistent with the energy transfer interpretation. Using these results and exploiting the 2-fold symmetry of the dimer, the chromophore chromophore distance R(DA) was calculated and found to be in the range 49 A < R(DA) < 63 A. Further evidence for energy transfer in Cph1 dimers was obtained from dilution experiments with PEB/PEB dimers: the lifetime was unchanged, but the anisotropy increased as the dimers dissociated with increasing dilution. These experiments allowed a rough estimate of 5 +/- 3 microM for the dimer dissociation constant. With the deletion mutant Cph1Delta2 that lacks the carboxy terminal histidine kinase domain less energy transfer was observed suggesting that in this mutant dimerization is much weaker. The carboxy terminal domain of Cph1 that is involved in intersubunit trans-phosphorylation and signal transduction thus plays a dominant role in the dimerization. The FRET method provides a sensitive assay to monitor the association of Cph1 monomers. PMID- 12741848 TI - Conformational dynamics of partially denatured myoglobin studied by time-resolved electrospray mass spectrometry with online hydrogen-deuterium exchange. AB - This study demonstrates the use of electrospray mass spectrometry in conjunction with rapid online mixing ("time-resolved" ESI-MS) for monitoring protein conformational dynamics under equilibrium conditions. The hydrogen/deuterium exchange (HDX) kinetics of mildly denatured myoglobin (Mb) at pD 9.3, in the presence of 27% acetonitrile, were studied with millisecond time resolution. Analytical ultracentrifugation indicates that the average protein compactness under these solvent conditions is similar to that of native holomyoglobin (hMb). The mass spectrum shows protein ions in a wide array of charge and heme binding states, indicating the presence of multiple coexisting conformations. The experimental approach used allows the HDX kinetics of all of these species to be monitored separately. A combination of EX1 and EX2 behavior was observed for hMb ions in charge states 7+ to 9+, which predominantly represent nativelike hMb in solution. The EX1 kinetics are biphasic, indicating the presence of two protein populations that undergo conformational opening events with different rate constants. The EX2 kinetics observed for nativelike hMb are biphasic as well. All other charge and heme binding states represent non-native protein conformations that are involved in rapid interconversion processes, thus leading to monoexponential EX2 kinetics with a common rate constant. Burst phase labeling for these non-native proteins occurs at 125 sites. In contrast, the nativelike protein conformation shows burst phase labeling only for 88 sites. A kinetic model is developed which is based on the assumption of three distinct (un)folding units in Mb. The model implies that the free energy landscape of the protein exhibits a major barrier. The crossing of this barrier is most likely associated with slow, cooperative opening/closing events of the heme binding pocket. Rapid conformational fluctuations on either side of the barrier give rise to the observed EX2 kinetics. Simulated HDX kinetics based on this model are in excellent agreement with the experimental data. PMID- 12741849 TI - Two Toc34 homologues with different properties. AB - The Toc34 isoforms are located in the outer envelope membrane of plastids. In pea, Toc34 functions as a GTP dependent receptor for preproteins, which is controlled by protein phosphorylation. Two members of this family are present in Arabidopsis thaliana, namely, atToc34 and atToc33. AtToc33 is phosphorylated, as is the homologue in P. sativum, while atToc34 is not. The phosphorylation of atToc33 occurs on serine 181. The highest affinity for dimerization was for the heterodimer between Toc33 and Toc34 in the absence of GTP or GDP. Both proteins, atToc33 and atToc34, bind GTP with significantly higher affinity than GDP and are able to hydrolyze GTP. The intrinsic GTP hydrolysis rate of both proteins is comparable. Hydrolysis is strongly stimulated in the presence of preproteins, which are in turn released upon GTP hydrolysis. Preprotein subclasses exist, which show a strong preference for either the atToc33 or the atToc34 receptor as revealed by GTP hydrolysis rate stimulation and receptor precursor dissociation constants. Detailed analysis of precursor recognition supports the model of a GTP hydrolysis regulated receptor ligand interaction. PMID- 12741850 TI - Identification of basic amino acid residues important for citrate binding by the periplasmic receptor domain of the sensor kinase CitA. AB - The sensor kinase CitA and the response regulator CitB of Klebsiella pneumoniae form the paradigm of a subfamily of bacterial two-component regulatory systems that are capable of sensing tri- or dicarboxylates in the environment and then induce transporters for the uptake of these compounds. We recently showed that the separated periplasmic domain of CitA, termed CitAP (encompasses residues 45 176 supplemented with an N-terminal methionine residue and a C-terminal hexahistidine tag), is a highly specific citrate receptor with a K(d) of 5.5 microM at pH 7. To identify positively charged residues involved in binding the citrate anion, each of the arginine, lysine, and histidine residues in CitAP was exchanged for alanine, and the resulting 17 muteins were analyzed by isothermal titration calorimetry (ITC). In 12 cases, the K(d) for citrate was identical to that of wild-type CitAP or slightly changed (3.9-17.2 microM). In one case (R98A), the K(d) was 6-fold decreased (0.8 microM), whereas in four cases (R66A, H69A, R107A, and K109A) the K(d) was 38- to >300-fold increased (0.2 to >1 mM). The secondary structure of the latter five proteins in their apo-form as deduced from far-UV circular dichroism (CD) spectra did not differ from the apo-form of wild-type CitAP; however, all of them showed an increased thermostability. Citrate increased the melting point (T(m)) of wild-type CitAP and mutein R98A by 6.2 and 9.5 degrees C, respectively, but had no effect on the T(m) of the four proteins with disturbed binding. Three of the residues important for citrate binding (R66, H69, and R107) are highly conserved in the CitA subfamily of sensor kinases, indicating that they might be involved in ligand binding by many of these sensor kinases. PMID- 12741851 TI - Analysis of mutations made during active synthesis or extension of mismatched substrates further define the mechanism of HIV-RT mutagenesis. AB - The effect of reverse transcriptase (RT) catalyzed mutations on continued extension of the nascent DNA chain was investigated. A system using the alpha-lac gene of beta-galactosidase as template and two sets of conditions was used. In one, RT was allowed to reassociate with the primer-template after falling off, while in a second RT was sequestered after dissociating. In the first condition, subsequent extension of errors that may have initially caused enzyme dissociation can occur. In the second, such errors would not be extended. Fully extended products were assayed by alpha-complementation to assess mutation frequency. A lower frequency in the latter scenario implies that some errors caused the polymerase to dissociate. Allowing only a single binding event lowered the mutation frequency of the products by about 1/2 suggesting that approximately 1 in 2 errors terminated synthesis. In other experiments, when added to a primer template with a terminal mismatch at the 3' end, RT dissociated from the template about 50-90% of the time (depending on mismatch type) rather than extending. Running start reactions indicated that extension was more likely if an actively synthesizing RT made the mutation. RT RNase H cleavage analysis showed that 3' mismatches weakened the association of RT with the primer-terminus. Taken together, these results suggest that an actively synthesizing RT enzyme that has just made a mistake is likely bound in a configuration that generally enhances extension of the mistake. This is in contrast to RTs that must bind to then extend mismatches. The importance of these findings with respect to the mechanism of mutagenesis is discussed. PMID- 12741852 TI - Biochemical basis of the constitutive coprotease activity of RecA P67W protein. AB - The mutation of Pro67 to Trp (P67W) in the Escherichia coli RecA protein results in reduced recombination and constitutive coprotease phenotypes. We examined the biochemical properties of this mutant in an effort to understand these altered behaviors. We find that RecA P67W protein can access single-stranded DNA (ssDNA) binding sites within regions of secondary structure more effectively than wild type protein, and binding to duplex DNA is both faster and more extensive as well. This mutant is also more effective than wild-type RecA protein in displacing SSB protein from ssDNA. An enhancement in SSB protein displacement has been shown previously for RecA441, RecA730, and RecA803 proteins, and similarly, this improved ability to displace SSB protein for RecA P67W protein correlates with an increased rate of association with ssDNA. As for the aforementioned mutant RecA proteins, we expect that this enhanced activity will allow RecA P67W protein to bind ssDNA naturally occurring in undamaged cells and to constitutively induce the SOS response. The DNA strand exchange activity of RecA P67W protein is also altered. Although the rate of duplex DNA uptake into joint molecules is increased compared to that of wild-type RecA protein, the resolution to the nicked circular dsDNA product is reduced. We suggest that either a limited amount of DNA strand reinvasion or a defect in DNA heteroduplex extension is responsible for the impaired recombination ability of this mutant protein. PMID- 12741853 TI - Biochemical characterization of a mutant RecA protein altered in DNA-binding loop 1. AB - The double substitution of Glu156 with Leu and Gly157 with Val in the Escherichia coli RecA protein results in a severely reduced level of recombination and constitutive coprotease behavior. Here we present our examination of the biochemical properties of this mutant protein, RecA N99, in an effort to understand its phenotype and the role of loop 1 (L1) in RecA function. We find that RecA N99 protein has reduced single-stranded DNA (ssDNA)-dependent ATP hydrolysis activity, which is not as sensitive to the presence of SSB protein as wild-type RecA protein. RecA N99 protein is also nearly unable to utilize duplex DNA as a polynucleotide cofactor for ATP hydrolysis, and it shows both a decreased rate of association with ssDNA and a diminished capacity to bind DNA in the secondary binding site. The mutant protein has a corresponding reduction in DNA strand exchange activity, which probably results in the decrease in recombination activity in vivo. The constitutive induction of the SOS response may be a consequence of the impaired ability to repair damaged DNA, resulting in unrepaired ssDNA which can act as a cofactor for the cleavage of LexA repressor. These findings point to an involvement of L1 in both the primary and secondary DNA binding sites of the RecA protein. PMID- 12741854 TI - Rat polymerase beta binds double-stranded DNA using exclusively the 8-kDa domain. Stoichiometries, intrinsic affinities, and cooperativities. AB - Analyses of the interactions of rat polymerase beta (rat pol beta) with a double stranded DNA have been performed using the quantitative fluorescence titration and fluorescence energy transfer techniques. The obtained results show that rat pol beta binds to dsDNA oligomers with the site-size of the enzyme-dsDNA complex n = 5 +/- 1 base pairs. The small site-size of the complex is a consequence of engagement of only the 8-kDa domain in intrinsic interactions with the dsDNA. This conclusion is directly supported by the fluorescence energy transfer between the single tryptophan residue on the 31-kDa domain and fluorescence acceptor located on the DNA. The dsDNA oligomer is bound at a distance of at least 55 A from the tryptophan, excluding the 31-kDa domain from any closed contact with the DNA. Moreover, in the complex with the dsDNA, the enzyme is bound in "open" conformational state. The intrinsic interactions are accompanied by a net release of about four to five ions. The net ion release is dominated by cations as a result of the exclusive engagement of the 8-kDa domain in interactions. Magnesium affects the net ion release through direct binding of Mg(2+) cations to the protein. Surprisingly, binding of rat pol beta to the dsDNA is characterized by strong positive cooperative interactions, a very different behavior from that previously observed for pol beta complexes with the ssDNA and gapped DNAs. Contrary to intrinsic affinities, cooperative interactions are accompanied by a net uptake of about three to five ions. Anions have a large contribution to the net ion uptake, indicating that cooperative interactions characterize protein protein interactions. The significance of these results for the pol beta functioning in damaged-DNA recognition processes is discussed. PMID- 12741855 TI - Peter V. Karpovich: transforming the strength paradigm. AB - One of the most important figures in the public's acceptance of weight training as an acceptable activity for athletes was Russian-born physician Peter V. Karpovich of Springfield College, Springfield, MA. Karpovich, like most early 20th-century educators, opposed weight training for athletes and held a low opinion of weightlifting as an activity in general. However, he became strength science's most eminent and visible advocate after witnessing a demonstration of weightlifting organized by Bob Hoffman of the York Barbell Company in 1940. Following that demonstration, Karpovich conducted several seminal studies that examined the bedrock beliefs on which the arguments normally cited against lifting were built-that it would make a person slow and inflexible-in short, muscle-bound. His research consistently revealed that those beliefs were in error. Later, he went on to collaborate with Jim Murray on the first science based book on the subject of strength training, Weight Training in Athletics, published in 1956. PMID- 12741856 TI - Validation of submaximal prediction equations for the 1 repetition maximum bench press test on a group of collegiate football players. AB - The purpose of the study was to determine the accuracy of 11 prediction equations in estimating the 1 repetition maximum (1 RM) bench press from repetitions completed by collegiate football players (N = 69) using 225 lb. The demographic variables race, age, height, weight, fat-free weight, and percent body fat were measured to determine whether these variables increased the accuracy of the prediction equations; race was the most frequently selected variable in the regression analyses. The validity of the prediction equations was dependent upon the number of repetitions performed, i.e., validity was higher when fewer repetitions were completed. Explained variability of 1 RM was slightly higher for all 11 equations when demographic variables were included. A new prediction equation was also developed using the number of repetitions performed and the demographic variables height and fat-free weight. PMID- 12741857 TI - Body composition in Division I football players. AB - This study assessed body composition of Division I football players (n = 69) and compared the findings with previously reported data to ascertain whether the increase in player total body mass that has been observed over the past 10 years has been accompanied by an increase in body fat. Body composition was determined by hydrostatic weighing and the measurement of skinfold thicknesses. Total body mass, skinfold thicknesses, and body fat were greater in the current players than in players in studies conducted in the early 1980s and early 1990s. Body fat varied significantly across playing position, with the defensive backs, offensive backs, and receivers being the leanest and the offensive linemen and tight ends the most fat. There was no significant relationship between body composition and playing year or scholarship status, nor were any differences observed between ethnic groups. Of important clinical relevance was the finding that the linemen (offensive, defensive) and tight ends were on average greater than 25% body fat, the borderline for obesity in this age group. Much of this fat was deposited in the abdominal region, a significant finding when one considers the high correlation between abdominal obesity and ischemic heart disease and stroke. The current findings suggest that more attention needs to be given to the nature of the increase in body mass being achieved by today's football player to minimize long-term negative health consequences, and the findings reemphasize the need identified in earlier studies of the importance of detraining programs for these athletes. PMID- 12741858 TI - Body composition relates poorly to performance tests in NCAA Division III football players. AB - We assessed body composition (height, body mass, body mass index, body fat by densitometry, fat mass, fat-free mass, and lean/fat ratio) and performance (10- and 40-yd sprints, pro shuttle run, vertical jump, sit and reach, and bench press) in 77 National Collegiate Athletic Association Division III football players. Data were analyzed by position and playing status. Significant differences (p 0.05). Number of repetitions performed at 72 hours was significantly higher than that in session 1 (10.2 +/- 1.4 reps in session 1 vs. 11.2 +/- 2.3 at 72 hours, p = 0.022) in the young 3-sets group, but not in the other groups. After 96 hours, only the young 7-sets group was found to be performing at a level approaching significance (10.3 +/- 1.2 reps in session 1 vs. 11.1 +/- 2.0 at 96 hours, p = 0.051). No significant difference was found between the young 3-sets and 7-sets groups at any time (p > 0.05). The young 3 sets group was found to be performing at a significantly higher level than the older group at 72 hours (11.2 +/- 2.3 reps in the younger vs. 9.9 +/- 1.7 in the older group, p = 0.008), a difference that also approached significance at 96 hours (p = 0.06). Large intersubject variability was observed at all time points. The results suggest that individual recovery testing before exercise prescription is practical, and this protocol may be sensitive to differences in training volume and subject age. PMID- 12741862 TI - A comparison of two stretching protocols on hip range of motion: implications for total daily stretch duration. AB - It is theorized that the total stretch time in a day is more important than the actual single stretch duration time. The purpose of this study was to compare 2 stretching protocols, keeping total stretching time equivalent. The 2 protocols were a 10-second duration stretch and a 30-second duration stretch. Although the stretch durations differed, the total stretching time over the course of a day was held constant at 2 minutes for both protocols. Participants were randomly assigned a protocol to each of their legs: subjects stretched 1 leg with the 10 second protocol and the opposite leg with the 30-second protocol. The 10-second stretch was repeated 6 times for a total of 1 minute; the 30-second protocol was repeated 2 times for a total of 1 minute. Stretching was performed twice daily (a total of 2 minutes each day) for 6 weeks. All stretching was performed to the hamstring muscles. Hip flexion measurements were recorded at pretest, 3-weeks, and 6-weeks. Subjects demonstrated significant gains in range of motion for hip flexion over the course of 6 weeks, p = 0.000. No differences existed between the 2 protocols. Range of motion gains were equal between the 2 stretching protocols. The common denominator was total stretch time for a day. Regardless of the duration of a single stretch, the key to improvement was the total daily stretch time. These findings are important as they allow clinicians and individuals to customize stretching protocols to meet individual needs. PMID- 12741863 TI - Effects of environmental cooling on force production in the quadriceps and hamstrings. AB - The purpose of this study was to determine the effects of environmental cooling on force production in the quadriceps and hamstring muscles. Ten men (mean +/- SD: age = 21.4 +/- 2.2 years, height = 168.5 +/- 35.9 cm, body mass = 78.0 +/- 6.4 kg) participated in this study. Each subject completed 2 sets of 10 maximal effort repetitions on a Cybex II isokinetic dynamometer at 3.14 rad x s(-1). Between sets, subjects sat in environmental temperatures of 20, 15, 10, or 5 degrees C for 40 minutes. A significant decrease (p 0.05). These findings provide evidence that hearing acuity and auditory function in young women do not change after short-term exposure to moderate-intensity exercise and loud music. Thus, listening to loud music with earphones during moderate-intensity exercise does not pose acute hearing health concerns for young physically fit adults with normal hearing. PMID- 12741867 TI - The effects of competition and the presence of an audience on weight lifting performance. AB - The purpose of this investigation was to examine the effects of the presence of an audience and competition on maximal weight lifting performance. Thirty-two recreationally trained participants (15 men, 17 women; 21 +/- 2.5 years) performed a 1 repetition maximum (1 RM) bench press during 3 different situations (coaction, competitive coaction, and audience condition). Subjects also completed the Activation-Deactivation Adjective Checklist Short Form following the 3 trials to measure arousal state during each of the 3 trials. Significant differences (p < 0.05) were found between competitive coaction and coaction trials as well as between audience and coaction trials. Both men and women demonstrated the highest performance in front of an audience (105 +/- 48 kg) followed by competition (103 +/- 46 kg) and then the coaction trial (93 +/- 43 kg). No significant difference in arousal was measured between trials. The data suggest that performing a maximal lift in the presence of an audience or in competition facilitates performance and support the self-presentation and self-awareness theories. Social facilitation effects should be controlled in research settings and may aid the performance of weight lifting activities during events or competition. PMID- 12741869 TI - Physiological cost of running while wearing spring-boots. AB - The purpose of the study was to investigate the physiological cost of running in spring-boots compared with running in running shoes at different speeds. During testing, subjects (n = 7) completed running trials while wearing spring-boots and running shoes. Three speed conditions (2.23, 2.68, and 3.13 m.s(-1)) were completed per shoe condition (i.e., spring-boots and running shoes). Rate of oxygen consumption (Vo(2)), heart rate (HR), rating of perceived exertion (RPE), and stride frequency were recorded for each condition. Order of shoe conditions was balanced, with speeds tested continuously from slow to fast. There was no difference in Vo(2), HR, or RPE between shoe conditions across speeds (p > 0.05). Stride frequency was lower during running in spring-boots vs. running shoes at each speed (speed of spring-boots vs. running shoes for 2.23 m x s(-1): 69.9 +/- 2.9 strides x min(-1) vs. 75.6 +/- 3.5 strides x min(-1); for 2.68 m x s(-1): 71.3 +/- 5.2 strides x min(-1) vs. 79.4 +/- 5.0 strides x min(-1); for 3.13 m x s(-1): 73.6 +/- 7.3 strides x min(-1) vs. 83.1 +/- 8.2 strides x min(-1); p < 0.05). Despite the added mass to the lower extremity and change in stride frequency during running in spring-boots, the physiological cost of running was similar to that of running in running shoes. Exercising while running in spring boots may provide less impact force with no change in running economy. PMID- 12741870 TI - Effectiveness of cycle cross-training between competitive seasons in female distance runners. AB - The purpose of this study was to examine whether substituting 50% of run training volume with cycling ("cross-training") would maintain 3,000-m race time and estimated Vo(2)max in competitive female distance runners during a 5-week recuperative phase. Eleven collegiate runners were randomly assigned to either the run training-only (R) group (n = 6) or the cycle training (R/C) group (n = 5), which cross-trained on alternate days. The groups trained daily at a reduced intensity (75-80% of maximum heart rate). Training volume was similar to the competitive season (40-50 mi x wk(-1)) except that cycling represented 50% of volume for the R/C group. On follow-up, 3,000-m time was 1.4% (9 seconds) slower in the R group and 3.4% (22 seconds) slower in the R/C group. No important change in estimated Vo(2)max was found for either group. It was concluded that cycle cross-training adequately maintained aerobic performance during the recuperative phase between the cross-country and track seasons, comparable to the primary sport of running. PMID- 12741871 TI - A method for predicting maximal strength in collegiate women athletes. AB - The purpose of this study was to develop a regression equation capable of accurately predicting a 1 repetition maximum bench press in collegiate women athletes. The findings of this study could benefit future women athletes by providing coaches and trainers with an easy method of determining maximum upper body strength in women athletes. Sixty-five University of Georgia NCAA Division 1 women athletes from 9 different sports were measured prior to the start of their season utilizing 2 repetition tests to fatigue (25 kg: REPS55; 31.8 kg: REPS70) and a 1 repetition maximum (1RM) bench press test in random order. Other independent variables that were used with a submaximal weight to predict 1RM were total body weight, lean body mass (LBM), height, and percent body fat. The variables of REPS70 and LBM were the best predictors of 1RM utilizing Pearson product correlations (r = 0.909, p = 0.000; r = 0.445, p = 0.000) and multiple regression results (R(2) = 0.834, p = 0.000) for this population. The results from this study indicate muscular endurance repetitions using an absolute weight of 31.8 kg in conjunction with LBM can be used to accurately predict 1RM bench press strength in collegiate women athletes. PMID- 12741872 TI - Evaluation of cardiovascular demands of game play and practice in women's ice hockey. AB - Preparation for the physical demands of competition often involves game simulation during practice. This paradigm is thought to promote physiological adaptations that enhance maximal performance. However, a mismatch between practice intensity and actual competition intensity may not provide adequate training to achieve optimal game-play fitness. The purpose of this study was to evaluate the effectiveness of practice in meeting the cardiovascular demands of a women's ice hockey game. Heart rate (HR) data from 11 U.S. National Women's Ice Hockey team members were collected (5-second intervals) during a game and a typical practice session. Data was normalized to individual HRmax determined during Vo(2)max testing. Working time was defined as a game shift or practice working interval. Mean working HR was greater during the game than the practice, 90 +/- 2% and 76 +/- 3% of HRmax, respectively (p < 0.05). Mean percent session time (game or practice) >90% HRmax was also longer during the game than the practice, 10.5 +/- 4.1% and 5.6 +/- 3.5% (p < 0.05), respectively. Mean session HR, percent time >80% HRmax, and mean resting HR were not different between game and practice (68 +/- 7% vs. 69 +/- 5%, 23.2 +/- 5.3% vs. 26.1 +/- 9.2%, and 59 +/ 8% vs. 56 +/- 5%, respectively). Elite women hockey players experience significantly greater cardiovascular load during game play than during practice. This mismatch in cardiovascular demand may prevent players from achieving "game shape," thus affecting competition play. PMID- 12741873 TI - Physiological responses using 2 high-speed resistance training protocols. AB - This study compared physiological responses to 2 high-speed resistance training (RT) protocols in untrained adults. Both RT protocols included 12 repetitions for the same 6 exercises, only differing in continuous (1 x 12) or discontinuous (2 x 6) mode. For discontinuous mode, there was a 15-second rest interval between sets. We hypothesized that the 2 x 6 protocol was less physiologically demanding than the 1 x 12 protocol. Fifteen untrained adults randomly performed the protocols on 2 different days while heart rate (HR), blood lactate (BL), rate of perceived exertion (RPE), and concentric phase mean power (CPMP) were measured. Significantly lower values (mean +/- SE) were seen with the discontinuous protocol for exercise HR (119 +/- 5 vs. 124 +/- 5 b x min(-1)), BL (5.7 +/- 0.5 vs. 6.7 +/- 0.3 mMol/L), and RPE (5.4 +/- 0.3 vs. 5.8 +/- 0.4) (p < 0.05). CPMP tended to be higher in the discontinuous protocol, especially for the 2 last repetitions. The discontinuous protocol was significantly less physiologically demanding, although similar or higher CPMP values were obtained. These findings may help foster long-term adherence to RT in untrained individuals. However, future studies are needed to compare physiological adaptations induced by these 2 RT protocols. PMID- 12741874 TI - Effect of active vs. passive recovery on repeat suicide run time. AB - This study was conducted to evaluate the difference between active and passive recovery methods during successive suicide runs by Division I women's collegiate basketball athletes (n = 14). Testing consisted of sprinting suicides on the basketball court using both traditional (short) and reverse-sequence (long) protocols. Two 90-second recovery methods were used, passive (standing still) and active (slow self-paced jogging). Although successive run time was reduced by a mean of 0.55 seconds after passive recovery relative to active, it did not reach significance (p = 0.09). Likewise, the difference between long and short line versions was nonsignificant (p = 0.41). Therefore, neither line sequence nor 90 second recovery technique appears to influence subsequent run time when performing 2 maximal-effort suicides. PMID- 12741875 TI - Effect of a submaximal half-squats warm-up program on vertical jumping ability. AB - The purpose of the current research was to study the effect of a warm-up program including submaximal half-squats on vertical jumping ability. Twenty physically active men participated in the study. Each subject performed 5 sets of half squats with 2 repetitions at each of the following intensities: 20, 40, 60, 80, and 90% of the 1 repetition maximum (1RM) load. Prior to the first set and immediately after the end of the last set, the subjects performed 2 countermovement jumps on a Kistler force platform; the primary goal was to jump as high as possible. The results showed that mean vertical jumping ability improved by 2.39% after the warm-up period. Subjects were then divided into 2 groups according to their 1RM values for the half-squat. Subjects with greater maximal strength ability improved their vertical jumping ability (4.01%) more than did subjects with lower maximal strength (0.42%). A warm-up protocol including half-squats with submaximal loads and explosive execution can be used for short-term improvements of vertical jumping performance, and this effect is greater in athletes with a relatively high strength ability. PMID- 12741876 TI - Kinetic analysis of complex training rest interval effect on vertical jump performance. AB - Complex training has been recommended as a method of incorporating plyometrics with strength training. Some research suggests that plyometric performance is enhanced when performed 3-4 minutes after the strength training set, whereas other studies have failed to find any complex training advantage when plyometrics are performed immediately after the strength training portion of the complex. The purpose of this study was to determine if there is an ergogenic advantage associated with complex training and if there is an optimal time for performing plyometrics after the strength training set. Subjects were 21 NCAA Division I athletes who performed a countermovement vertical jump, a set of 5 repetitions maximum (5 RM) squats, and 5 trials of countermovement vertical jump at intervals of 10 seconds and 1, 2, 3, and 4 minutes after the squat. Jump height and peak ground reaction forces were acquired via a force platform. The pre-squat jump performance was compared with the post-squat jumps. Repeated measures ANOVA determined a difference (p 0.05) was found comparing subsequent jumps (0.72-0.76 m) to the pre-squat condition (0.74 m). When comparing high to low strength individuals, there was no effect on jump performance following the squat (p > 0.05). In conclusion, complex training does not appear to enhance jumping performance significantly and actually decreases it when the jump is performed immediately following the strength training set; however, a nonsignificant trend toward improvement seemed to be present. Therefore to optimize jump performance it appears that athletes should not perform jumps immediately following resistance training. It may be possible that beyond 4 minutes of recovery performance could be enhanced; however, that was not within the scope of the current study. PMID- 12741877 TI - Energy cost of the ACSM single-set resistance training protocol. AB - The purpose of this study was (a) to assess the energy cost and intensity of a single-set resistance training (RT) protocol conducted according to the recent ACSM guidelines and (b) to compare obtained values to those recently reported as eliciting health benefits via endurance-based physical activity (PA). Twelve subjects, mean age 26.7 +/- 3.8 years, performed 1 set of a 15 repetition maximum (15 RM) for each of 8 RT exercises. Metabolic data were collected via a portable calorimetric system. Training intensity in metabolic equivalents (METS) was 3.9 +/- 0.4 for men and 4.2 +/- 0.6 for women (not significant). Total energy was 135.20 +/- 16.6 kcal for men and 81.7 +/- 11.1 kcal for women (p < 0.008). We concluded that the ACSM single-set, 8-exercise RT protocol is a feasible alternative for achieving moderate-intensity (3-6 METS) PA, but it is not sufficient to achieve a moderate amount (150-200 kcal) of PA. PMID- 12741879 TI - Thermal responses and body fluid balance of competitive male swimmers during a training session. AB - Thermoregulatory and body fluid balance (BFB) responses of competitive swimmers were studied during a typical interval training session under natural field conditions. Subjects were 9 males (18.0 +/- 1.7 years; VO(2)max = 3.8 +/- 0.9 L x min(-1)) who covered 9,000 m in 180 minutes in an outdoor pool (mean water temperature = 26.8 +/- 0.3 degrees C; mean wet bulb globe temperature = 29.8 +/- 2.8 degrees C). Mean body weight (BWt) decreased by 1.8 +/- 0.5 kg (P < 0.05), and rectal temperature increased by 1.0 +/- 1.0 degrees C (P < 0.05). Volitional water intake (WI) (0.1 +/- 0.2 kg) did not maintain BFB (-0.5 kg per hour) and plasma volume decreased 10.7 +/- 5.4%. During a typical training session, swimmers experienced significant body fluid losses, and WI was not enough to prevent involuntary dehydration. The magnitude of the fluid losses (2.5% of BWt) was sufficient to compromise convective thermoregulation because of the decreased plasma volume. Hence, to prevent involuntary dehydration, swimmers should be encouraged to consume an amount of fluids that equals losses throughout the training sessions. PMID- 12741878 TI - Effect of Roman chair exercise training on the development of lumbar extension strength. AB - The purpose of this study was to determine the effect of 45 degrees Roman chair exercise training on the development of lumbar extension strength. Fifteen healthy volunteers (9 women, 6 men) were recruited from a university setting and were randomly assigned to 1 of 2 groups. One group (n = 9) performed progressive resistance back extension exercise on a 45 degrees Roman chair once weekly for 12 weeks. Training consisted of one set of 8-20 dynamic repetitions to volitional exhaustion using hand-held metal plates for additional resistance. The other group did not train (control, n = 6). Peak isometric lumbar extension torque was measured on a lumbar extension dynamometer before and after the 12-week program. Following training, peak isometric lumbar extension torque did not increase for the Roman chair group (before: 224.0 +/- 134.1 N x m; after: 240.3 +/- 137.4 N x m; p > 0.05) compared with the control group (before: 175.6 +/- 68.9 N x m; after: 178.2 +/- 69.9 N x m; p > 0.05), despite an increase in dynamic exercise load. PMID- 12741881 TI - Adding weights to stretching exercise increases passive range of motion for healthy elderly. AB - Stretching exercise is effective for increasing joint range of motion (ROM). However, the Surgeon General's Report and the American College of Sports Medicine cite a lack of studies identifying strategies capable of increasing the effectiveness of stretching exercise. This investigation evaluated adding modest weight (0.45-1.35 kg) to a stretching exercise routine (Body Recall [BR]) on joint ROM. Forty-three subjects ages 55-83 years participated in 1 of 2 training groups, BR, BR with weights (BR+W), or a control group (C). ROM was evaluated at the neck, shoulder, hip, knee, and ankle before and after 10 weeks of exercise. Using ANCOVA, significant differences (p < 0.01) were observed for right and left cervical rotation, hip extension, ankle dorsiflexion, ankle plantar flexion, and shoulder flexion. Post hoc analysis revealed that cervical rotation (left and right), hip extension, and ankle dorsiflexion for BR+W subjects differed significantly from BR and C (p < 0.01). Significant differences with shoulder flexion and ankle plantar flexion were found for both BR and BR+W in comparison to C (p < 0.01). Results indicate that addition of weights enhanced the effectiveness of stretching exercise for increasing joint ROM with 4 of the 6 selected measurements. Thus, a modest intensity exercise program that is within the reach of most elderly may significantly affect joint ROM and flexibility. PMID- 12741880 TI - A survey of the scientific data and training methods utilized by collegiate strength and conditioning coaches. AB - The purpose of this study was to determine the extent to which scientific research influences college strength and conditioning coaching practices and to determine the training methods utilized. A total of 321 surveys were mailed to Division I strength and conditioning coaches, and the response rate was 42.7% (137 of 321 surveys). Results indicate that all subjects held a baccalaureate degree, the majority in a human performance-related field, and that 75% were Certified Strength and Conditioning Specialist (CSCS) certified. The respondents' most widely utilized professional resources were the Strength and Conditioning Journal (94%) and other collegiate coaches and programs (93%). Forty-seven percent of respondents indicated that other collegiate coaches and their programs were the most important sources of knowledge outside of formal education. The majority indicated that they used a periodization protocol (93%) utilizing multiple sets (97%), plyometrics (90%), explosive movements (88%), and Olympic lifts (85%). Respondents tend to rely on sources of information that may not be defined as scientific, as evidenced by the low priority given to peer-reviewed literature. Respondents also tend to employ the methods they utilized as athletes. Reliance on these sources may not take advantage of advances made through scientific research in exercise physiology, biomechanics, and more specifically the area of strength and conditioning. PMID- 12741882 TI - Changes evaluated in soccer-specific power endurance either with or without a 10 week, in-season, intermittent, high-intensity training protocol. AB - The purpose of this study was to evaluate changes in soccer-specific power endurance of 34 female high school soccer players throughout a season either with or without an intermittent, high-intensity exercise protocol. Thirty-four female high school soccer players were tested prior to the 2000 fall season and again 10 weeks later. The tests included an abridged 45-minute shuttle test (LIST), hydrostatic weighing, vertical jump, 20-m running-start sprint, and 30-second Wingate test. The experimental group (EG; n = 17, age 16.5 +/- 0.9 years) completed a 10-week in-season plyometric, resistive training, and high-intensity anaerobic program. The control group (n = 17, age 16.3 +/- 1.4 years) completed only traditional aerobic soccer conditioning. Statistical significance was set at alpha < 0.05. The experimental group showed significant improvements in the LIST (EG = delta 394 seconds +/- 124 seconds), 20-m sprint (EG = Delta-0.10 seconds +/ 0.10 seconds), increase in fat-free mass (EG = delta 1.14 kg +/- 1.22 kg), and decreases in fat mass (EG = Delta-1.40 kg +/- 1.47 kg) comparing pre- to postseason. This study indicates that a strength and plyometric program improved power endurance and speed over aerobic training only. Soccer-specific power endurance training may improve match performance and decrease fatigue in young female soccer players. PMID- 12741883 TI - Intermatch variation of match activity in elite Italian soccer referees. AB - The aim of this study was to examine how match activities, specifically high intensity activity (HIA; activities performed at speeds faster than 18.1 km x h( 1)), vary among matches in elite soccer referees, because variations in match performance can potentially guide physical conditioning regimens for these referees. Fourteen Italian soccer referees were observed during 65 first-division matches a minimum of 3 and a maximum of 6 times each. For comparison, shorter matches (SM) and longer matches (LM) for each referee were identified. Mean (+/ SD) SM and LM total distances were 10,949 +/- 1,095 m and 12,303 +/- 666 m (p < 0.001), respectively. SM and LM were not different in term of duration (p > 0.05). During LM, referees spent 44% less time standing still (p < 0.01) and covered 10% more distance at low intensity than they did in SM (p < 0.05). No HIA variation was observed between SM and LM (p > 0.05). For the elite referee, work rate is increased without affecting HIA. PMID- 12741884 TI - Early phase changes by concurrent endurance and strength training. AB - To compare regimens of concurrent strength and endurance training, 26 male basketball players were matched for stature, body composition, and physical activity level. Subjects completed different training programs for 7 weeks, 4 days per week. Groups were as follows: (a) the strength group (S; n = 7) did strength training; (b) the endurance group (E; n = 7) did endurance training; (c) the strength and endurance group (S + E; n = 7) combined strength and endurance training; and (d) the control group (C; n = 5) had no training. The S + E group showed greater gains in Vo(2)max than the E group did (12.9% vs. 6.8%), whereas the S group showed a decline (8.8%). Gains were noted in strength and vertical jump performance for the S + E and S groups. The S + E group had better posttraining anaerobic power than the S group did (6.2% vs. 2.9%). No strength, power, or anaerobic power gains were present for the E and C groups. We conclude that concurrent endurance and strength training is more effective in terms of improving athletic performance than are endurance and strength training apart. PMID- 12741885 TI - Muscle fiber characteristics of competitive power lifters. AB - To examine the skeletal muscle characteristics of power lifters, 5 competitive power lifters (PL; X +/- SE; age = 31.0 +/- 1.5 years, squat = 287.7 +/- 15.7 kg, bench press = 170.5 +/- 17.7 kg, and deadlift = 284.2 +/- 7.5 kg) and 5 untrained control subjects (CON; age = 27.3 +/- 3.3 years) served as subjects. Isokinetic squat force and power was greater (p < 0.05) for the PL at all bar velocities (0.20, 0.82, and 1.43 m;pd s(-1)), as was vertical jump height and estimated power. Muscle biopsies from the vastus lateralis m. revealed significant differences for percent fiber type (PL, IIA = 45.5 +/- 1.6%, IIB = 1.3 +/- 0.8%; CON, IIA = 33.4 +/- 3.1%, IIB = 12.0 +/- 2.4%); percent fiber type area (PL, IIA = 51.8 +/- 1.6%, IIB = 1.3 +/- 0.8%; CON, IIA = 43.5 +/- 3.4%, IIB = 12.4 +/- 2.6%); and percent myosin heavy chain isoform (PL, IIa = 59.5 +/- 6.1%; CON, 46.5 +/- 2.5%). Muscle fiber characteristics were significantly correlated (r = +/- 0.61) with numerous strength and power measures for the PL. These data illustrate the muscle fiber characteristics necessary for the maximal force production requirements of power lifting. PMID- 12741886 TI - Effect of pre-exhaustion exercise on lower-extremity muscle activation during a leg press exercise. AB - The purpose of this study was to investigate the effect of pre-exhaustion exercise on lower-extremity muscle activation during a leg press exercise. Pre exhaustion exercise, a technique frequently used by weight trainers, involves combining a single-joint exercise immediately followed by a related multijoint exercise (e.g., a knee extension exercise followed by a leg press exercise). Seventeen healthy male subjects performed 1 set of a leg press exercise with and without pre-exhaustion exercise, which consisted of 1 set of a knee extension exercise. Both exercises were performed at a load of 10 repetitions maximum (10 RM). Electromyography (EMG) was recorded from the rectus femoris, vastus lateralis, and gluteus maximus muscles simultaneously during the leg press exercise. The number of repetitions of the leg press exercise performed by subjects with and without pre-exhaustion exercise was also documented. The activation of the rectus femoris and the vastus lateralis muscles during the leg press exercise was significantly less when subjects were pre-exhausted (p < 0.05). No significant EMG change was observed for the gluteus maximus muscle. When in a pre-exhausted state, subjects performed significantly (p < 0.001) less repetitions of the leg press exercise. Our findings do not support the popular belief of weight trainers that performing pre-exhaustion exercise is more effective in order to enhance muscle activity compared with regular weight training. Conversely, pre-exhaustion exercise may have disadvantageous effects on performance, such as decreased muscle activity and reduction in strength, during multijoint exercise. PMID- 12741887 TI - Results of the survey of the quality assurance for commercially prepared microbiology media. Update from the College of American Pathologists Microbiology Surveys Program (2001). AB - CONTEXT: Beginning in 1985, state-of-the-art surveys by the College of American Pathologists (CAP) led to M22-A guidelines by the National Committee for Clinical Laboratory Standards (NCCLS) for quality assurance (QA) of various commercially prepared isolation and diagnostic microbiology media. Some medium types were declared exempt from routine QA testing by each laboratory user, but manufacturer QA was maintained and specified in great detail. No update has occurred in more than 15 years as more complex media have been introduced into clinical microbiology practice. OBJECTIVE: To reassess the quality of commercially prepared microbiology media in the United States for the revision of NCCLS document M22-A2. Methods.-A questionnaire was designed to conform to that used by the CAP in 1985 and 1988, and was updated by lists of 52 recently introduced diagnostic media. Queries included details of laboratory QA in the last 12 months, such as number of lots tested, number of items in a lot, number of items in a quality control (QC) sample, number of lots failed, and reasons for lot failure. All CAP Microbiology Surveys participants received the document in late 2001. RESULTS: Data from nearly 300 000 media lots representing 32.7 million medium items were received from more than 3000 CAP Surveys subscribers. Lot raw data failure rates for all media ranged from 0.10% to 9.87% (average, 1.01%). Failures with media having a significant QC experience (> or =1000 lots or > or =100 000 items) adjusted for QC strain-based failures (so-called extrapolated failure rate) ranged from 0.04% to 1.34% (average, 0.40%). The threshold rate for QA exemption from 1985 analyses was expanded to 0.50% or less and now allows exemption of 27 media evaluated in this study. Local laboratory QA must remain in force (M22-A2) for the other 25 products. CONCLUSIONS: Generated QA failure results from the CAP Microbiology Surveys supplement recent results of the NCCLS M22 Subcommittee that will lead to more practical media QA guidelines and could significantly decrease costs for clinical microbiology laboratories through focusing QC on a smaller number of the most at-risk diagnostic products. PMID- 12741888 TI - Pathology resident attitudes and opinions about pathologists' assistants. AB - CONTEXT: Changes in health care economics and organization have resulted in increased use of nonphysician providers in most health care settings. Attitudinal acceptance of nonphysician providers is important in the current health care environment. OBJECTIVES: To obtain descriptive information regarding pathology resident attitudes and opinions about pathologists' assistants in anatomic pathology practice and to assess the implications of resident attitudes and opinions for pathology practice and training. DESIGN: A self-administered, mailed, voluntary, anonymous questionnaire was distributed to a cross-sectional sample of pathology residents in the United States (2531 pathology residents registered as resident members of one of the national pathology professional organizations). The questionnaire contained (1) items relating to resident demographics and program characteristics, (2) Likert-scale response items containing positive and negative statements about pathologists' assistants, (3) a multiple-choice item related to pathologists' assistants scope of practice, and (4) an open-ended item inviting additional comments. Both quantitative and qualitative analysis of responses was performed. RESULTS: The overall response rate was 19.4% (n = 490); 50% of the respondents were women, and 77% reported use of pathologists' assistants in their program. Most respondents were 25 to 35 years old and in postgraduate years 3 through 5 of their training, and most were located in the Midwestern United States. The majority of residents expressed overall positive attitudes and opinions about pathologists' assistants and felt that pathologists' assistants enhanced resident training by optimizing resident workload. A minority (10%-20%) of residents expressed negative attitudes or opinions about pathologists' assistants. Additionally, some residents reported a lack of knowledge about pathologists' assistants' training or roles. CONCLUSIONS: Increased resident education and open discussion concerning pathologists' assistants may be beneficial for optimizing resident attitudes about and training experiences with pathologists' assistants. PMID- 12741889 TI - Colorectal carcinoma nodal staging. Frequency and nature of cytokeratin-positive cells in sentinel and nonsentinel lymph nodes. AB - CONTEXT: Nodal staging accuracy is important for prognosis and selection of patients for chemotherapy. Sentinel lymph node (SLN) mapping improves staging accuracy in breast cancer and melanoma and is being investigated for colorectal carcinoma. OBJECTIVE: To assess pathologic aspects of SLN staging for colon cancer. DESIGN: Sentinel lymph nodes were identified with a dual surgeon pathologist technique in 51 colorectal carcinomas and 12 adenomas. The frequency of cytokeratin (CK)-positive cells in mesenteric lymph nodes, both SLN and non SLN, was determined along with their immunohistochemical characteristics. RESULTS: The median number of SLNs was 3; the median number of total nodes was 14. The CK-positive cell clusters were detected in the SLNs of 10 (29%) of 34 SLN negative patients. Adjusted per patient, SLNs were significantly more likely to contain CK-positive cells than non-SLNs (P <.001). Cell clusters, cytologic atypia, and/or coexpression of tumor and epithelial markers p53 and E-cadherin were supportive of carcinoma cells. Single CK-positive cells only, however, could not be definitively characterized as isolated tumor cells; these cells generally lacked malignant cytologic features and coexpression of tumor and epithelial markers and in 2 cases represented mesothelial cells with calretinin immunoreactivity. Colorectal adenomas were associated with a rare SLN CK-positive cell in 1 (8%) of 12 cases. CONCLUSIONS: Sentinel lymph node staging with CK immunohistochemical analysis for colorectal carcinomas is highly sensitive for detection of nodal tumor cells. Cohesive cell clusters can be reliably reported as isolated tumor cells. Single CK-positive cells should be interpreted with caution, because they may occasionally represent benign epithelial or mesothelial cells. PMID- 12741890 TI - Concept-match medical data scrubbing. How pathology text can be used in research. AB - CONTEXT: In the normal course of activity, pathologists create and archive immense data sets of scientifically valuable information. Researchers need pathology-based data sets, annotated with clinical information and linked to archived tissues, to discover and validate new diagnostic tests and therapies. Pathology records can be used for research purposes (without obtaining informed patient consent for each use of each record), provided the data are rendered harmless. Large data sets can be made harmless through 3 computational steps: (1) deidentification, the removal or modification of data fields that can be used to identify a patient (name, social security number, etc); (2) rendering the data ambiguous, ensuring that every data record in a public data set has a nonunique set of characterizing data; and (3) data scrubbing, the removal or transformation of words in free text that can be used to identify persons or that contain information that is incriminating or otherwise private. This article addresses the problem of data scrubbing. OBJECTIVE: To design and implement a general algorithm that scrubs pathology free text, removing all identifying or private information. METHODS: The Concept-Match algorithm steps through confidential text. When a medical term matching a standard nomenclature term is encountered, the term is replaced by a nomenclature code and a synonym for the original term. When a high-frequency "stop" word, such as a, an, the, or for, is encountered, it is left in place. When any other word is encountered, it is blocked and replaced by asterisks. This produces a scrubbed text. An open-source implementation of the algorithm is freely available. RESULTS: The Concept-Match scrub method transformed pathology free text into scrubbed output that preserved the sense of the original sentences, while it blocked terms that did not match terms found in the Unified Medical Language System (UMLS). The scrubbed product is safe, in the restricted sense that the output retains only standard medical terms. The software implementation scrubbed more than half a million surgical pathology report phrases in less than an hour. CONCLUSIONS: Computerized scrubbing can render the textual portion of a pathology report harmless for research purposes. Scrubbing and deidentification methods allow pathologists to create and use large pathology databases to conduct medical research. PMID- 12741891 TI - A regional centralized microbiology service in Calgary for the rapid diagnosis of malaria. AB - CONTEXT: A regional centralized laboratory service for the rapid diagnosis of malaria was implemented 3 years ago in May 1999 within the Division of Microbiology, Calgary Laboratory Services. OBJECTIVE: To describe the design and performance of this unique microbiology laboratory service. DESIGN: Blood specimens must arrive at the central laboratory within 2 hours of collection. Thin blood smears are read and reported from suspected acute cases within 1 hour of receipt, 24 hours per day, 7 days a week, by trained and experienced microbiology technologists. All positive malaria smears are reviewed by a medical microbiologist and confirmed by polymerase chain reaction at a reference laboratory. SETTING: Calgary Laboratory Services provides integrated laboratory services to the Calgary Health Region, an urban area of more than 1 million people. MAIN OUTCOME MEASURES: Performance of the service has been continuously monitored by measuring preanalytic and analytic test turnaround times, test accuracy, clinical relevance, and the results of proficiency testing. RESULTS: More than 90% of blood specimens for malaria from community locations have consistently arrived within 2 hours of collection, and hospitals have reached this target within the past year. Although polymerase chain reaction was more sensitive at detecting the presence of malaria, the expert microscopists were as accurate at determining the type of Plasmodium infection. More than 95% of all positive smear results are consistently reported within 2 hours of receipt of a blood specimen. CONCLUSIONS: Implementation of a regional centralized microbiology service has improved our ability to make a rapid and accurate diagnosis of malaria in this region. PMID- 12741892 TI - Microsatellite instability in colorectal carcinoma. The comparison of immunohistochemistry and molecular biology suggests a role for hMSH6 [correction of hMLH6] immunostaining. AB - CONTEXT: Microsatellite instability (MSI) due to defective mismatch repair (MMR) genes has been reported in the majority of colorectal tumors from patients with hereditary nonpolyposis colorectal cancer syndrome and in 10% to 15% of sporadic colorectal cancers. The identification of cancers associated with MSI requires classical molecular testing as the gold standard. OBJECTIVE: The aim of this study was to evaluate the role of immunohistochemistry with antibodies directed against 4 MMR proteins as a screening tool for carcinomas with MSI. METHODS: In this study, 204 formalin-fixed, paraffin-embedded colorectal carcinomas were examined for MMR protein expression (hMLH1, hMSH2, hMSH6, and hPMS2) and analyzed for MSI (MSI-H indicates at least 2 of 6 markers affected). These results were correlated with histopathologic parameters. RESULTS: Immunohistochemical analysis revealed that loss of expression of at least 1 protein was present in 17% of cases. One hundred percent of carcinomas that showed high instability (MSI-H) showed loss of expression of hMLH1, hMSH2, or hMSH6. Loss of expression of 2 proteins was present in 59.4% of MSI-H cases, with only 2 combinations, namely, hMLH1/hPMS2 and hMSH2/hMSH6. Isolated loss of hMSH6 expression was present in 2 MSI-H cases. CONCLUSIONS: These findings confirm that examination of MMR protein expression by immunohistochemistry is a simple method to diagnose colorectal cancer with MSI. Our data suggest that the study of hMSH6 may be useful, in addition to hMLH1 and hMSH2. Moreover, immunohistochemistry could represent a screening method with which to direct research on the mutations of MMR genes observed in hereditary nonpolyposis colorectal cancer syndrome. PMID- 12741893 TI - Evaluation of immunohistochemistry and multiple-level sectioning in sentinel lymph nodes from patients with breast cancer. AB - CONTEXT: Previous investigations on sentinel lymph node biopsies have demonstrated their importance in nodal staging of patients with breast cancer. However, sentinel node biopsy in breast cancer is currently a controversial procedure and continues to provoke debate. OBJECTIVES: We designed our study to determine the usefulness of a standard protocol for evaluating sentinel lymph node metastases and to assess the value of sentinel node biopsy as the only procedure in nodal staging in breast cancer patients. MATERIALS AND METHODS: A retrospective analysis of 84 breast cancer patients with sentinel node biopsies, who also underwent axillary dissection, was conducted using a standard protocol (3 levels of immunohistochemical stains for keratin and 2 levels of hematoxylin eosin (HE) stains on the first 3 negative lymph nodes). RESULTS: Hematoxylin eosin staining identified 20 patients (23.8%) with sentinel node metastases. The remaining 64 negative patients (76.1%) were tumor free on sentinel lymph nodes at level 1 HE. Additional immunohistochemical stains for keratin and HE stains on specimens from these 64 patients showed an additional 5 patients (7.8%) to be positive for lymph node micrometastases (<2 mm). The total percentage of cases with sentinel lymph node metastases detected by HE staining and immunohistochemistry was 29.7%. Of the remaining 59 cases that were negative on HE and immunohistochemistry, axillary dissection revealed 3 cases that had metastases in the axillary lymph nodes. The false-negative rate was 10.7%. The concordance rate between sentinel lymph nodes and axillary lymph nodes was 96.4%. The sensitivity was 89% and specificity was 100%. CONCLUSION: Immunohistochemistry and multiple-level sectioning increased detection of metastases by 7.8% in sentinel lymph nodes. Caution should be used in accepting sentinel node biopsy alone as the only procedure for staging due to a high false negative rate (10.7%). A predictive value of 96.4% confirms that sentinel lymph node biopsy is most likely to contain metastatic carcinoma. Sentinel lymph node examination with the protocol we describe, combined with axillary dissection, increased the yield of metastatic disease by identifying 8 additional cases of nodal metastatic disease (an increase of 28%), as compared to standard axillary nodal dissection and single-section sentinel lymph node examination alone. PMID- 12741894 TI - Rapid polymerase chain reaction-based confirmation of cat scratch disease and Bartonella henselae infection. AB - CONTEXT: Cat scratch disease (CSD) commonly occurs secondary to Bartonella henselae infection, and the diagnosis has traditionally been made by microscopic findings, the identification of organisms by cytochemistry, and clinical history. However, cytochemical analysis tends to be very difficult to interpret, and histology alone may be insufficient to establish a definitive diagnosis of CSD. OBJECTIVE: To demonstrate the presence of B henselae in tissue suspected of involvement by CSD, using a novel polymerase chain reaction (PCR) assay. DESIGN: Isolates of B henselae (American Tissue Culture Collection 49793) and Afipia felis (American Tissue Culture Collection 49714) were cultured on blood agar and buffered charcoal yeast extract agar, respectively. DNA was isolated from these organisms and from formalin-fixed, paraffin-embedded tissue sections with involvement by CSD (8 patients). Negative controls included water, human placental tissue, and lymph node specimens from 6 patients with reactive lymphoid hyperplasia and from 2 patients with granulomatous lymphadenitis. A primer complementary to B henselae citrate synthase gltA gene sequence was designed to perform a seminested PCR amplification. For restriction fragment length polymorphism analysis, PCR products were digested by TaqI restriction enzyme and analyzed by gel electrophoresis. RESULTS: Seminested PCR analysis of the cultured isolates of B henselae, but not of A felis, showed specific amplification. However, nonnested PCR did not provide consistently positive results in tissue sections with CSD. Therefore, we used a seminested PCR, which revealed positivity in all of the cases with clinicopathologic diagnoses of CSD. None of the negative controls showed positivity. Restriction enzyme provided confirmation of the specific PCR amplification of the B henselae sequence. CONCLUSIONS: Since the amplification product has a low molecular size (<200 base pairs), this assay is useful for detection of B henselae in formalin-fixed, paraffin-embedded tissues. The seminested PCR protocol described here can be used for rapid and reliable confirmation of B henselae in samples that are histologically suggestive of CSD. PMID- 12741895 TI - Disappearance of meconium pigment in placental specimens on exposure to light. AB - CONTEXT: Meconium discharge has been associated with fetal distress and poor neonatal outcome; thus, its presence is of clinical importance. OBJECTIVE: Loss of meconium pigment in histologic sections from light exposure has been described. We sought to confirm this finding and to measure this loss quantitatively. DESIGN: Sections of umbilical cord, fetal membranes, and fetal surface from 11 grossly meconium-stained placentas were processed swiftly to minimize light exposure. Two serial sections from each block were cut and stained; one set was reviewed immediately, and the other was exposed to 8 hours of direct fluorescent lighting. Each site and exposure was scored for pigment intensity (0, no staining; 1, weak expression; and 2, moderate/strong expression) and number of meconium-laden macrophages per 10 high-power fields (HPF). Results were compared on the same specimen using the chi(2) and the paired-samples t test. RESULTS: The maximum meconium macrophage count was 13.2/10 HPF in the unexposed sections versus 6.1/10 HPF in the exposed sections (P <.001). Unexposed sections varied from 1+ to 2+ intensity, while exposed sections were all 1+ or negative (P <.001). CONCLUSION: Exposure to fluorescent laboratory lights for 8 hours resulted in a significant loss in the intensity and number of identifiable meconium macrophages in histologic sections. These findings have important implications in the handling of placental specimens, and we recommend that care be taken to minimize exposure to laboratory lights during processing. PMID- 12741896 TI - Prevalence and incidence of cytoplasmic yellow bodies in thyroid neoplasms. AB - CONTEXT: Cytoplasmic yellow bodies are a common and frequent histologic finding in hyalinizing trabecular adenoma of the thyroid gland, a morphologically distinctive neoplasm, and are visible in fine-needle aspiration biopsy of the tumor. OBJECTIVE: To determine the prevalence and frequency of cytoplasmic yellow bodies in common thyroid tumors. DESIGN: Microscopic slides of random cases of papillary carcinoma (61 cases), follicular adenoma (27 cases), and Hurthle cell adenoma (12 cases) were searched for cytoplasmic yellow bodies. SETTING AND PATIENTS: Slides were from patients who had undergone surgery at Mayo Clinic, Rochester, Minn, and were obtained from the Mayo Clinic Tissue Registry. MAIN OUTCOME MEASURE: Presence or absence of cytoplasmic yellow bodies. RESULTS: Cytoplasmic yellow bodies were found in papillary carcinoma (62%), follicular carcinoma (22%), and Hurthle cell adenoma (83%) but were very infrequent numerically in each tumor type. CONCLUSIONS: Cytoplasmic yellow bodies may be present in papillary carcinoma and in follicular and Hurthle cell adenomas, but because they are uncommon in these tumors, they are unlikely to be found in fine needle aspiration biopsy smears. Therefore, cytoplasmic yellow bodies are a useful cytomorphologic indicator of hyalinizing trabecular adenoma of the thyroid gland. PMID- 12741897 TI - Comparison of direct selective versus nonselective agar media plus LIM broth enrichment for determination of group B streptococcus colonization status in pregnant women. AB - CONTEXT: Group B streptococcus (GBS) is the most common cause of early-onset neonatal sepsis in developed countries, and determination of the GBS colonization status in pregnant patients near term is essential for the provision of prophylactic measures to prevent early-onset disease. OBJECTIVES: To determine if GBS recovery rates and/or result turnaround times for vaginal or combined vaginal/rectal swab specimens from pregnant patients near term are enhanced if swabs are inoculated initially onto selective versus nonselective agar media, in addition to the standard Centers for Disease Control and Prevention method. DESIGN: Prospective laboratory analysis. SETTING: Urban health region/centralized diagnostic microbiology laboratory. PATIENTS: Pregnant women presenting for routine obstetrical care and collection of vaginal or combined vaginal/rectal swab specimens for GBS testing at 35 to 37 weeks' gestation. INTERVENTION: Culture of specimens directly onto selective (5% sheep blood with colistin and nalidixic acid) or nonselective (5% sheep blood) agar media, in addition to LIM broth enrichment and terminal subculture. MAIN OUTCOME MEASURES: Group B streptococcus recovery rate and culture result turnaround time. RESULTS: A total of 639 specimens were tested, with 128 (20%) positive for GBS. Sixty-three isolates were recovered on direct agar media at 24 hours, of which 16 (12.5%) were isolated on selective plates only. An additional 38 isolates were recovered at 48 hours from direct plates. Twenty-seven (21.1%) isolates that failed to grow on direct plates were recovered from the LIM broth subculture only. Three (2.3%) isolates not recovered from LIM broths were detected at 48 hours on the direct selective (2 isolates) and nonselective (1 isolate) agar plates. A 24-hour result turnaround time was achieved for 63 (49.2%) and 47 (36.7%) of the 128 culture positive specimens for direct selective and nonselective plates, respectively (chi2 = 76.63, P <.001). CONCLUSIONS: Use of direct selective agar media, in addition to LIM broth enrichment, for the determination of the GBS colonization status in pregnant patients near term results in decreased turnaround time for reporting positive results. PMID- 12741898 TI - Comparison of voice-automated transcription and human transcription in generating pathology reports. AB - CONTEXT: Software that can convert spoken words into written text has been available since the early 1980s. Early continuous speech systems were developed in 1994, with the latest commercially available editions having a claimed accuracy of up to 98% of speech recognition at natural speech rates. OBJECTIVES: To evaluate the efficacy of one commercially available voice-recognition software system with pathology vocabulary in generating pathology reports and to compare this with human transcription. To draw cost analysis conclusions regarding human versus computer-based transcription. DESIGN: Two hundred six routine pathology reports from the surgical pathology material handled at St Joseph's Healthcare, Hamilton, Ontario, were generated simultaneously using computer-based transcription and human transcription. The following hardware and software were used: a desktop 450-MHz Intel Pentium III processor with 192 MB of RAM, a speech quality sound card (Sound Blaster), noise-canceling headset microphone, and IBM ViaVoice Pro version 8 with pathology vocabulary support (Voice Automated, Huntington Beach, Calif). The cost of the hardware and software used was approximately Can 2250 dollars. RESULTS: A total of 23 458 words were transcribed using both methods with a mean of 114 words per report. The mean accuracy rate was 93.6% (range, 87.4%-96%) using the computer software, compared to a mean accuracy of 99.6% (range, 99.4%-99.8%) for human transcription (P <.001). Time needed to edit documents by the primary evaluator (M.A.) using the computer was on average twice that needed for editing the documents produced by human transcriptionists (range, 1.4-3.5 times). The extra time needed to edit documents was 67 minutes per week (13 minutes per day). CONCLUSIONS: Computer-based continuous speech-recognition systems in pathology can be successfully used in pathology practice even during the handling of gross pathology specimens. The relatively low accuracy rate of this voice-recognition software with resultant increased editing burden on pathologists may not encourage its application on a wide scale in pathology departments with sufficient human transcription services, despite significant potential financial savings. However, computer-based transcription represents an attractive and relatively inexpensive alternative to human transcription in departments where there is a shortage of transcription services, and will no doubt become more commonly used in pathology departments in the future. PMID- 12741899 TI - Automation of gross photography using a remote-controlled digital camera system. AB - CONTEXT: Conventional gross photography requires a series of tedious and time consuming steps, including taking, developing, labeling, sorting, filing, and tracking numerous photographs. OBJECTIVE: To describe how to automate the gross photographic process by way of controlling a digital camera remotely. DESIGN: After defining the requirements of automation regarding gross photography, a remote control board, foot switch, barcode system, and image retrieval system were devised. SETTING: The surgical pathology laboratory of a university medical center with a commercially available megapixel digital camera. RESULTS: The digital camera zoom and shutter were controlled remotely by a foot switch. A large portion of the gross photographic process, including specimen number labeling, image downloading, labeling, sorting, filing, and tracking, were automated. In addition, the elimination of several manual specimen-processing steps, along with not having to wait for the developing and mounting of conventional 35-mm film, reduced the entire time span required in conventional gross photography from 2 to 5 days, to a few minutes. It was also possible to review the gross images at the time of microscopic sign-out. CONCLUSIONS: The automation of gross photography using a remote-controlled digital camera changes the conventional gross workflow markedly. We found use of a remote-controlled gross photography system to be practical, convenient, and efficient. PMID- 12741900 TI - Gastric toxoplasmosis in a patient with acquired immunodeficiency syndrome. A case report and review of the literature. AB - Toxoplasmosis is a common opportunistic pathogen in patients with acquired immunodeficiency syndrome (AIDS). It usually presents with ocular, central nervous system, or pulmonary disease. Gastric toxoplasmosis is uncommon in AIDS patients, especially in the absence of central nervous system manifestations. In the few reported cases, patients have presented with abdominal pain and other digestive complaints that usually are attributed to the more common gastrointestinal manifestations of human immunodeficiency virus infection. We describe a 49-year-old man with AIDS who presented with abdominal pain, diarrhea, dry cough, and systemic symptoms and was diagnosed with toxoplasmosis by a gastric biopsy. PMID- 12741902 TI - Xanthomatous pseudotumor. An unusual postchemotherapy phenomenon in breast cancer. AB - Neoadjuvant chemotherapy has become an integral part of the treatment for locally advanced breast cancer. It facilitates tumor resectability and also provides an opportunity for the assessment of therapeutic response and prognosis. We report a case in which a large primary breast carcinoma was significantly reduced in size clinically and replaced by a mass lesion that was composed almost entirely of foamy histiocytes. This peculiar phenomenon is described in detail, together with a brief review of the other known postchemotherapy histologic features that include tumor necrosis, tumor cell cytoplasmic vacuolation and marked nuclear atypia, accompanying chronic inflammatory cellular infiltrate, fibrosis, and ductal-lobular atrophy. PMID- 12741901 TI - Amyloid A gastrointestinal amyloidosis associated with idiopathic retroperitoneal fibrosis. Report of a rare autopsy case and review of the literature. AB - We report a rare autopsy case of secondary gastrointestinal amyloid A (AA) amyloidosis associated with idiopathic retroperitoneal fibrosis (IRF) in a 67 year-old woman. Masses were identified around the aorta and inferior vena cava in her abdomen. Histologically, plasma cell infiltration was observed within fibrotic areas. Because no specific cause for the inflammatory mass was apparent, we diagnosed it as IRF. Steroid therapy, which usually reduces IRF masses, proved ineffective, and malabsorption syndrome developed 4 years later. On autopsy, amyloid protein was present systemically in the vascular walls of several organs, and deposition was highest in the gastrointestinal mucosa. Amyloid protein was identified as AA type, strongly suggesting that the amyloidosis was secondarily induced by IRF. To our knowledge, only 2 other cases of IRF-associated amyloidosis have been reported. Two of the 3 patients of these cases were women who showed resistance to steroid therapy. PMID- 12741903 TI - Diffuse fibrous pseudotumor of the testicular tunics associated with an inflamed hydrocele. AB - Fibrous pseudotumors of the testicular tunics and paratesticular soft tissue are uncommon lesions. They typically arise as painless scrotal masses that may be associated with a hydrocele or history of trauma or infection. Although these lesions are clinically worrisome for a malignant neoplasm, they are thought to be reactive in nature, since they are composed of dense fibrous tissue with interspersed bland fibroblasts and myofibroblasts and mixed inflammatory cells. Once excised, these lesions behave in a benign fashion. Typically, these masses are multinodular, but in rare cases they are diffuse, bandlike myofibroblastic proliferations that encase the testis and are termed fibromatous periorchitis. Herein, we describe a 57-year-old man with a diffuse fibrous pseudotumor/fibromatous periorchitis that encased the left testis and adnexa and arose in close apposition to an inflamed hydrocele. PMID- 12741904 TI - Clear cell carcinoid tumor of the gallbladder. A case without von Hippel-Lindau disease. AB - A golden yellow polyp was detected in the gallbladder of a 64-year-old man who presented with epigastric pain. The lesion was composed of clear polygonal cells arranged in a trabecular and glandular pattern. The tumor invaded through the wall into the perimuscular subserosal layer. Immunohistochemical stains showed that neoplastic cells were positive for chromogranin A, synaptophysin, somatostatin, gastrin, and pancreatic polypeptide and negative for glucagon, serotonin, insulin, S100 protein, and inhibin. This tumor resembles the recently described clear cell endocrine tumors of the gallbladder and pancreas that are associated with von Hippel-Lindau disease. Our patient, however, had neither personal nor family history indicative of von Hippel-Lindau disease. Furthermore, published accounts of clear cell endocrine tumors in von Hippel-Lindau disease describe immunoreactivity for inhibin; the current case was negative for the disease. There may be a subtype of clear cell carcinoid tumor not associated with von Hippel-Lindau disease, which is characterized by its lack of immunoreactivity against inhibin. PMID- 12741905 TI - Indeterminate cell histiocytosis. Clinical and pathologic study in a pediatric patient. AB - Indeterminate cell histiocytosis is a rare disorder involving altered homing mechanisms of the cutaneous histiocytic/dendritic system. It has been described predominantly in adults, with less than a dozen cases in children. A 13-year-old adolescent girl presented with a 4-year history of asymptomatic erythematous nodules and plaques, measuring from 1 to 5 cm in diameter, that were located mainly on the trunk and proximal portions of her limbs. A skin biopsy showed dermal diffuse infiltration of histiocytic cells. Most of the histiocytic cells were strongly positive for S100 protein. No Birbeck granules were found. Treatment with topical steroid was ineffective. After 6 months of pure coal tar and 5% 5-fluorouracil cream, an almost total clearing of lesions was observed. An accurate diagnosis of this condition is mandatory in order to avoid unnecessary treatments. Conservative management is also discussed. PMID- 12741906 TI - Pathologic quiz case: a 2-year-old boy with a hereditary bleeding disorder. PMID- 12741907 TI - Pathologic quiz case: a child with facial and proximal limb weakness. PMID- 12741908 TI - Pathologic quiz case: brown macule on the thigh of a 71-year-old woman. PMID- 12741909 TI - Pathologic quiz case: a 31-year-old woman with menometrorrhagia. PMID- 12741910 TI - Gastric anthrax. PMID- 12741911 TI - Amebic colitis in an asymptomatic patient. PMID- 12741917 TI - Pathologic quiz case: a 23-year-old man with a solitary red calf lesion. PMID- 12741918 TI - Pathologic quiz case: a young woman with an indurated skin plaque. PMID- 12741919 TI - Pathologic quiz case: right lower quadrant pain in a 2-year-old boy. PMID- 12741920 TI - Pathologic quiz case: a large ovarian mass in an otherwise healthy 32-year-old woman. PMID- 12741921 TI - Pathologic quiz case: a child with clumsy gait. PMID- 12741923 TI - Pathologic quiz case: a man with an asymptomatic left testicular mass. PMID- 12741922 TI - Pathologic quiz case: a 66-year-old man with thyrotoxicosis. PMID- 12741924 TI - Pathologic quiz case: upper aerodigestive tract mass in a 90-year-old woman. PMID- 12741925 TI - Pathologic quiz case: newborn with a subcutaneous facial mass. PMID- 12741926 TI - Pathologic quiz case: a deep-seated tumor of the shoulder. PMID- 12741927 TI - Pathologic quiz case: a 56-year-old woman with a recurring neck mass. PMID- 12741928 TI - Pathologic quiz case: a 15-year-old adolescent girl with supraclavicular lymphadenopathy and pleural and pericardial effusions. PMID- 12741929 TI - Why have asthma action plans failed the consumer test? PMID- 12741930 TI - The SARS epidemic: lessons for Australia. PMID- 12741931 TI - Managing atrial fibrillation--redrawing a line in the sand. PMID- 12741932 TI - Prevalence of asthma and asthma action plans in South Australia: population surveys from 1990 to 2001. AB - OBJECTIVES: To assess trends in the prevalence of self-reported doctor-diagnosed asthma, associated asthma related morbidity, and the uptake of written asthma action plans in South Australia, 1990-2001. DESIGN, SETTING AND PARTICIPANTS: Surveys by telephone interview of the South Australian population between 1990 and 2001, and interview of participants in their own homes by trained health interviewers. MAIN OUTCOME MEASURES: Asthma prevalence, percentage of patients with written action plans, and asthma associated morbidity. RESULTS: The reported prevalence of doctor-diagnosed asthma has increased from 8% (95% CI, 6.4%-9.6%) in 1990 to 12.8% (95% CI, 11.4%-14.2%) in 2001. Morbidity, as measured by wakening at night (daily or weekly) and days lost from normal activities because of asthma, has remained constant over the decade. The percentage of patients with written asthma action plans increased to a peak of 42.3% (95% CI, 40.3%-44.3%) in 1995, but then declined to 22.2% (95% CI, 20.7%-23.7%) in 2001. CONCLUSIONS: The prevalence of asthma has increased while morbidity has remained constant, indicating that the burden of asthma has increased. The associated decline in the percentage of patients with asthma action plans in recent years is cause for concern. PMID- 12741933 TI - The effect of dexamethasone on the longevity of syringe driver subcutaneous sites in palliative care patients. AB - OBJECTIVE: To assess the effect of adding 1 mg dexamethasone to syringe drivers on the viability time of subcutaneous cannulation sites in palliative care patients. DESIGN: Prospective, double-blind, randomised, controlled trial in which patients received half their daily infused medications plus 1 mg dexamethasone in 1 mL saline through one subcutaneous site (test site) and the other half of their medications plus 1 mL saline through another symmetrically placed site (control site). PARTICIPANTS AND SETTING: Palliative care patients from the inpatient units at two hospices, recruited between 1999 and 2002. MAIN OUTCOME MEASURE: Difference in time that the test and control sites remained viable. RESULTS: 38 patients consented and were randomised. Twenty did not complete the trial because their participation in the study finished before either site broke down. Eighteen patients either partially completed (at least one site broke down) or fully completed (both sites broke down) the trial. In these 18 patients, test sites lasted 3.6 days longer than control sites (95% CI, 1.5-5.8 days; P = 0.002). Twelve patients fully completed the trial. In this group, test sites lasted 3.9 days longer than control sites (95% CI, 0.6-7.2 days; P = 0.025). CONCLUSIONS: The addition of 1 mg dexamethasone to syringe drivers significantly extends the viability time of subcutaneous cannulation sites in palliative care patients. PMID- 12741934 TI - Respiratory morbidity in central Australian Aboriginal children with alveolar lobar abnormalities. AB - OBJECTIVES: To describe the short-term outcomes in Aboriginal children admitted to hospital with radiological alveolar lobar changes; and determine whether predischarge chest radiography can predict respiratory morbidity found at follow up. DESIGN, PARTICIPANTS, SETTING: Prospective cohort study of Aboriginal children admitted to Alice Springs Hospital between October 2000 and April 2001 with alveolar lobar abnormalities (area of consolidation, > or = 1 cm) on chest radiographs. Participants were to have a predischarge radiograph and be followed up for 12 months. MAIN OUTCOME MEASURES: Comorbidities, follow-up rate, and new respiratory disease found at follow-up. RESULTS: Of 113 children hospitalised with radiological alveolar lobar changes, 109 were Aboriginal. Their median age was 1.8 years (range, 0.2 months-13.3 years), and 124 episodes were recorded. Comorbidities were common in these children (anaemia, 51.5%; suppurative otitis media, 37.3%). The follow-up rate one year after admission was 83.1% of episodes. New treatable chronic respiratory morbidity was found in 20 (25.6%) of the 78 children with completed follow-up. Predischarge chest radiographs were predictive of all chronic respiratory morbidity when they showed no or minimal resolution (0 20% resolution) (relative risk, 7.43; 95% CI, 2.07-26.60). CONCLUSIONS: Central Australian Aboriginal children admitted to hospital with alveolar changes on chest radiographs have a substantial burden of chronic respiratory illness, and should be clinically followed up for early detection and management of chronic respiratory morbidity. A predischarge radiograph is useful, and patients whose radiograph shows no or minimal resolution should have a follow-up x-ray film. PMID- 12741937 TI - Surgical training--a personal Koori journey. PMID- 12741936 TI - Inflammation and vascular endothelial activation in an Aboriginal population: relationships to coronary disease risk factors and nutritional markers. AB - OBJECTIVE: To describe the levels of inflammation and vascular endothelial activation in an Aboriginal community, and the relationship of these factors to coronary heart disease (CHD) risk factors and markers of nutritional quality. DESIGN AND PARTICIPANTS: A cross-sectional survey of 95 women and 76 men participating in a chronic-disease prevention program. SETTING: A remote Aboriginal community in Western Australia in 1996. MAIN OUTCOME MEASURES: Concentrations of markers of inflammation (C-reactive protein [CRP]) and vascular endothelial activation (soluble E-selectin [sE-selectin]); presence of metabolic syndrome; concentrations of diet-derived antioxidants. RESULTS: Participants exhibited very high plasma concentrations of CRP (mean, 5.4 mg/L; 95% CI, 4.6-6.3 mg/L) and sE-selectin (mean, 119 ng/mL; 95% CI, 111-128 ng/mL). Both CRP and sE selectin concentrations were significantly higher in the presence of the metabolic syndrome. There were significant inverse linear relationships between concentrations of CRP and plasma concentrations of the antioxidants lycopene, beta-carotene, cryptoxanthin and retinol. Even stronger inverse associations were evident between concentrations of sE-selectin and lycopene, beta-carotene, cryptoxanthin and lutein. CONCLUSIONS: Vascular inflammation and endothelial activation may be important mediators of elevated CHD risk in Aboriginal people. Inadequate nutrition and physical inactivity may contribute to this process. PMID- 12741938 TI - Indigenous health: it's time for a change. PMID- 12741939 TI - Pneumococcal meningitis masquerading as subarachnoid haemorrhage. AB - A 43-year-old woman taking warfarin for past venous thrombosis presented with 4 days of flu-like symptoms and deterioration in level of consciousness. Computed tomography suggested subarachnoid haemorrhage, and magnetic resonance imaging showed widespread cerebral infarcts. However, these seemed out of proportion to the amount of haemorrhage, and lumbar puncture revealed meningitis caused by Streptococcus pneumoniae. PMID- 12741941 TI - Daniel's message for doctors. PMID- 12741940 TI - Caring for the dying: the doctor as healer. PMID- 12741942 TI - The plague within: an Australian doctor's experience of SARS in Hong Kong. PMID- 12741943 TI - Working in partnership with support services in the era of the "new genetics". AB - Patient care in the "new genetics" era encompasses not only the diagnosis of a genetic condition or risk, but also managing the psychosocial, familial and ethical sequelae. Partnerships between the medical professional and expert clinical genetics services, support groups, registries and genetics education services provide a framework for this management. More than 750 Australian support groups assist individuals and families with genetic conditions through contact with peers, information and education resources for patients and professionals, practical advice about coping and advocacy. PMID- 12741944 TI - Enhanced chlamydia surveillance indicates more screening needed. PMID- 12741945 TI - Amoebic appendicitis. PMID- 12741946 TI - The clinical utility of routine urinalysis in pregnancy. PMID- 12741947 TI - Pap smear participation rates, primary healthcare and Indigenous women. PMID- 12741948 TI - How long should drug treatment of depression last? PMID- 12741951 TI - Access block: problems and progress. PMID- 12741952 TI - Inappropriate use of hospital emergency departments. PMID- 12741953 TI - Differential tissue and enzyme inhibitory effects of the vasopeptidase inhibitor omapatrilat in the rat. AB - Vasopeptidase inhibitors simultaneously inhibit angiotensin-converting enzyme (ACE) and neutral endopeptidase (NEP). The present study characterized the tissue distributions of ACE and NEP, and assessed the effects of the vasopeptidase inhibitor omapatrilat on ACE and NEP in rat tissues. In vivo ACE and NEP inhibition was studied by in vitro autoradiography and using the ACE inhibitor radioligand (125)I-MK351A and the NEP inhibitor radioligand (125)I-RB104 in rats that received oral omapatrilat (40 mg x day(-1) x kg(-1)) for 3 days. In vitro autoradiography was used to examine the distribution of ACE and NEP in the kidney, aorta, heart, adrenal gland, lung, intestine, liver, spleen and brain, and to assess enzyme inhibition after oral omapatrilat. Omapatrilat inhibited plasma ACE and increased plasma renin activity (P <0.01). Tissue ACE was inhibited by 70-95% (P <0.01), except in the brain, where ACE was not inhibited. NEP was inhibited by 87% in the kidney and by 20-40% in atria, aorta, adrenal gland, lung, liver and intestine; it was not inhibited in the brain, the ventricle or the spleen. Omapatrilat is a potent vasopeptidase inhibitor that significantly inhibits tissue ACE and NEP, with the degree of inhibition varying according to the enzyme and the tissue under assessment. The degree and site of tissue enzyme inhibition by vasopeptidase inhibitors may be relevant to end-organ protection as well as to the side-effect profiles of these agents. PMID- 12741954 TI - Characterization of SMOC-2, a modular extracellular calcium-binding protein. AB - We have isolated the novel gene SMOC-2, which encodes a secreted modular protein containing an EF-hand calcium-binding domain homologous to that in BM-40. It further consists of two thyroglobulin-like domains, a follistatin-like domain and a novel domain found only in the homologous SMOC-1. Phylogenetic analysis of the calcium-binding domain sequences showed that SMOC-1 and -2 form a separate group within the BM-40 family. The human and mouse SMOC-2 sequences are coded for by genes consisting of 13 exons located on chromosomes 6 and 17, respectively. Analysis of recombinantly expressed protein showed that SMOC-2 is a glycoprotein with a calcium-dependent conformation. Results from Northern blots and reverse transcription PCR revealed a widespread expression in many tissues. PMID- 12741955 TI - Training-induced adaptation of oxidative phosphorylation in skeletal muscles. AB - Muscle training/conditioning improves the adaptation of oxidative phosphorylation in skeletal muscles to physical exercise. However, the mechanisms underlying this adaptation are still not understood fully. By quantitative analysis of the existing experimental results, we show that training-induced acceleration of oxygen-uptake kinetics at the onset of exercise and improvement of ATP/ADP stability due to physical training are mainly caused by an increase in the amount of mitochondrial proteins and by an intensification of the parallel activation of ATP usage and ATP supply (increase in direct stimulation of oxidative phosphorylation complexes accompanying stimulation of ATP consumption) during exercise. PMID- 12741956 TI - Zinc-fingers and homeoboxes (ZHX) 2, a novel member of the ZHX family, functions as a transcriptional repressor. AB - Zinc-fingers and homeoboxes (ZHX) 1 is a transcription factor that interacts with the activation domain of the A subunit of nuclear factor-Y (NF-YA). Using a yeast two-hybrid system, a novel ubiquitous transcription factor ZHX2 as a ZHX1 interacting protein was cloned. ZHX2 consists of 837 amino acid residues and contains two zinc-finger motifs and five homeodomains (HDs) as well as ZHX1. The mRNA is expressed among various tissues. ZHX2 not only forms a heterodimer with ZHX1, but also forms a homodimer. Moreover, ZHX2 interacts with the activation domain of NF-YA. Further analysis revealed that ZHX2 is a transcriptional repressor that is localized in the nuclei. Since ZHX2 shares a number of properties in common with ZHX1, we conclude that all these come under the ZHX family. The minimal functional domains of ZHX2 were then characterized. The dimerization domain with both ZHX1 and ZHX2 is the region containing HD1, the domain that interacts with NF-YA is the HD1 to HD2 region, the repressor domain is the HD1 to a proline-rich region. Lastly, using an immunoprecipitation assay, we showed that ZHX2 intrinsically interacts with NF-YA in HEK-293 cells and that ZHX2 represses the promoter activity of the cdc25C gene stimulated by NF-Y in Drosophila Schneider line 2 cells. Thus the ZHX family of proteins may participate in the expression of a number of NF-Y-regulated genes via a more organized transcription network. PMID- 12741957 TI - A functional study on polymorphism of the ATP-binding cassette transporter ABCG2: critical role of arginine-482 in methotrexate transport. AB - Overexpression of the ATP-binding cassette transporter ABCG2 reportedly causes multidrug resistance, whereas altered drug-resistance profiles and substrate specificity are implicated for certain variant forms of ABCG2. At least three variant forms of ABCG2 have been hitherto documented on the basis of their amino acid moieties (i.e., arginine, glycine and threonine) at position 482. In the present study we have generated those ABCG2 variants by site-directed mutagenesis and expressed them in HEK-293 cells. Exogenous expression of the Arg(482), Gly(482), and Thr(482) variant forms of ABCG2 conferred HEK-293 cell resistance toward mitoxantrone 15-, 47- and 54-fold, respectively, as compared with mock transfected HEK-293 cells. The transport activity of those variants was examined by using plasma-membrane vesicles prepared from ABCG2-overexpressing HEK-293 cells. [Arg(482)]ABCG2 transports [(3)H]methotrexate in an ATP-dependent manner; however, no transport activity was observed with the other variants (Gly(482) and Thr(482)). Transport of methotrexate by [Arg(482)]ABCG2 was significantly inhibited by mitoxantrone, doxorubicin and rhodamine 123, but not by S octylglutathione. Furthermore, ABCG2 was found to exist in the plasma membrane as a homodimer bound via cysteinyl disulphide bond(s). Treatment with mercaptoethanol decreased its apparent molecular mass from 140 to 70 kDa. Nevertheless, ATP-dependent transport of methotrexate by [Arg(482)]ABCG2 was little affected by such mercaptoethanol treatment. It is concluded that Arg(482) is a critical amino acid moiety in the substrate specificity and transport of ABCG2 for certain drugs, such as methotrexate. PMID- 12741958 TI - Regioselectivity of glucosylation of caffeic acid by a UDP glucose:glucosyltransferase is maintained in planta. AB - Caffeic acid is a phenylpropanoid playing an important role in the pathways leading to lignin synthesis and the production of a wide variety of secondary metabolites. The compound is also an antioxidant and has potential utility as a general protectant against free radicals. Three glucosylated forms of caffeic acid are known to exist: the 3- O - and 4- O -glucosides and the glucose ester. This study describes for the first time a glucosyltransferase [UDP glucose:glucosyltransferase (UGT)] that is specific for the 3-hydroxyl, and not the 4-hydroxyl, position of caffeic acid. The UGT sequence of Arabidopsis, UGT71C1, has been expressed as a recombinant fusion protein in Escherichia coli, purified and assayed against a range of substrates in vitro. The assay confirmed that caffeic acid as the preferred substrate when compared with other hydroxycinnamates, although UGT71C1 also exhibited substantial activity towards flavonoid substrates, known to have structural features that can be recognized by many different UGTs. The expression of UGT71C1 in transgenic Arabidopsis was driven by the constitutive cauliflower mosaic virus 35 S (CaMV35S) promoter. Nine independent transgenic lines were taken to homozygosity and characterized by Northern-blot analysis, assay of enzyme activity in leaf extracts and HPLC analysis of the glucosides. The level of expression of UGT71C1 was enhanced considerably in several lines, leading to a higher level of the corresponding enzyme activity and a higher level of caffeoyl-3- O -glucoside. The data are discussed in the context of the utility of UGTs for natural product biotransformations. PMID- 12741960 TI - Pilot study to determine safety and short-term outcome of percutaneous cryoablation in the treatment of localized prostate cancer (stages T1C-T3C NO MO). AB - OBJECTIVE: To assess the effectiveness of cryoablation in localized prostate cancer in our hands. Early end points are biopsy at 5 months and PSA at 12 months follow-up. METHODS: Percutaneous transperineal cryoablation was performed on 30 patients. A urethral warming catheter was used. All cases were newly diagnosed with no prior radiotherapy, or neoadjuvant hormones. A single freeze cycle was used in the first 10 cases and two cycles in the next 20 cases. RESULTS: Twenty nine of 30 cases are eligible for follow-up. Six of the first 10 patients had negative biopsies, the 4 positive patients have had repeat cryoablation and only 1 patient remains biopsy positive. In cases 11-30, there were two patients with positive biopsies - both retreated. At this time, 27/29 patients (93%) have a negative biopsy after one (23 patients) or two (6 patients) treatments. At 6 months, 20/29 patients (69%) have undetectable PSA; 25/29 have PSA less than 1 ng/ml. At 12 months, 17/23 cases have an undetectable PSA (74%). Complications were minimal; 3 cases (10%) suffered urethral sloughing requiring TURP. CONCLUSION: Early results are very encouraging, showing 93% negative biopsy rate after one (23 patients) or two (6 patients) treatments; PSA is undetectable in 74% (17/23 patients) at 1 year. Early results of cryoablation for prostate cancer are encouraging. PMID- 12741961 TI - Survey of consecutive prostate cancer patients attending the Toronto Sunnybrook Regional Cancer Centre (TSRCC). AB - OBJECTIVE: To survey a sample of consecutive new prostate cancer patients attending the multidisciplinary clinic at the Toronto Sunnybrook Regional Cancer Centre (TSRCC). To assess the reason for their initial investigation, what treatment they received, the reason for their referral, their opinion and expectations prior to their referral to the TSRCC. To compare their choice of treatment with the reason for their referral, and the treatment recommended prior to their visit to the TSRCC. PMID- 12741962 TI - An unusual cause for granulomatous inflammation of the penis. AB - We report a highly unusual type of inflammatory reaction secondary to the insertion of needle in the penis with formation of a nodule of 2.1 cm in diameter, histologically mimicking lymphogranuloma venereum. PMID- 12741963 TI - Bilateral transitional cell carcinoma of the kidney: a case report. AB - Bilateral nephrectomy for transitional cell carcinoma of the renal pelvis is a rare procedure. We report two cases of bilateral nephrectomy for transitional cell carcinoma. Both patients were placed on dialysis. A review of the literature revealed three other cases of transitional cell carcinoma treated with bilateral nephrectomies. Based on our experience and the review of current literature, we recommend a waiting period of 2 years on dialysis post-operatively before proceeding to transplantation. PMID- 12741959 TI - The spatiotemporal expression pattern of the bone morphogenetic protein family in rat ovary cell types during the estrous cycle. AB - In the mammalian ovary, great interest in the expression and function of the bone morphogenetic protein (BMP) family has been recently generated from evidence of their critical role in determining folliculogenesis and female fertility. Despite extensive work, there is a need to understand the cellular sites of expression of these important regulatory molecules, and how their gene expression changes within the basic ovary cell types through the cycle. Here we have performed a detailed in situ hybridization analysis of the spatial and temporal expression patterns of the BMP ligands (BMP-2, -3, -3b, -4, -6, -7, -15), receptors (BMPR IA, -IB, -II), and BMP antagonist, follistatin, in rat ovaries over the normal estrous cycle. We have found that: i) all of the mRNAs are expressed in a cell specific manner in the major classes of ovary cell types (oocyte, granulosa, theca interstitial, theca externa, corpora lutea, secondary interstitial, vascular and ovary surface epithelium); and ii) most undergo dynamic changes during follicular and corpora luteal morphogenesis and histogenesis. The general principle to emerge from these studies is that the developmental programs of folliculogenesis (recruitment, selection, atresia), ovulation, and luteogenesis (luteinization, luteolysis) are accompanied by rather dramatic spatial and temporal changes in the expression patterns of these BMP genes. These results lead us to hypothesize previously unanticipated roles for the BMP family in determining fundamental developmental events that ensure the proper timing and developmental events required for the generation of the estrous cycle. PMID- 12741964 TI - Primary re-alignment of traumatic ruptured urethra with the aid of a wireloop. AB - A technique for endoscopic primary re-alignment of ruptured urethra is described. It is safe, simple and may be accomplished under neuroleptic or local anesthesia. Morbidity is low and hospitalization minimal. Any residual stricture can be easily managed endoscopically or by simple dilation. PMID- 12741965 TI - EDITORIAL. PMID- 12741966 TI - Molecular basis of androgen blockade in prostate cancer. AB - The molecular basis for antiandrogen therapy is reviewed. This article covers the structure and function of the androgen receptor, including the relationship between mutations in the receptor and the acquisition of androgen independent growth. Implications for anti-androgen therapy are addressed. PMID- 12741967 TI - Steroidal versus nonsteroidal antiandrogens in the management of prostate cancer. AB - The four antiandrogens currently available (cyproterone acetate, flutamide, nilutamide and bilcalutamide) are reviewed relative to their structure, pharmacology and clinical effect. PMID- 12741968 TI - Does maximal androgen blockade (MAB) improve survival? A critical appraisal of the evidence. AB - The conflicting data surrounding the benefit of total androgen blockade compared to monotherapy for metastatic prostate cancer is reviewed. The recent Lancet meta analysis is critically appraised. A sensitivity analysis of the studies is presented. PMID- 12741969 TI - Cavitary pulmonary metastases in transitional cell carcinoma. AB - Cavitating metastatic lesions in the lungs as a manifestation of metastatic transitional cell carcinoma (TCC) is rare. We herein present a case of multiple cavitary pulmonary metastases secondary to TCCF of the left ureter and bladder. PMID- 12741970 TI - Does a second bladder diverticulum require excision if the first contains a bladder tumor? AB - The multiplicity of bladder diverticula is well documented, yet review of the literature revealed only one reported case of cancer occurring in more than one diverticulum. To our knowledge, we report on the first long-term follow-up of a remaining bladder diverticulum after partial cystectomy for a transitional cell carcinoma in a contralateral diverticulum. The case report suggests that conservative management may be preferred for bladder diverticula not involved with tumor and routine prophylactic diverticulectomy may not be necessary. PMID- 12741971 TI - Sirenomelia: urological abnormalities in a surviving mermaid. AB - We report on a surviving four-year-old female with sirenomelia. She was born with the features of fused lower extremities, imperforate anus, colon atresia, pelvic and sacral dysplasia. Despite these severe anomalies, she had functional renal units. Maldevelopment of the genitourinary tract in patients with severe variant of caudal regression syndrome is usually incompatible with life. The case report includes the most current imaging studies of her unusual pelvic anomalies which poses a reconstructive challenge to the pediatric surgeon. PMID- 12741972 TI - EDITORIAL. PMID- 12741973 TI - Analysis of gland volume effect on the performance of PSA density in early detection of non-palpable, isoechoic prostate cancer. AB - OBJECTIVE: To reassess PSA density (PSAD) in the detection of non-palpable, isoechoic prostate cancer and to analyze the effect of potentially inaccurate non planimetric transrectal ultrasound volume estimates on the diagnostic performance of this diagnostic tool. METHODS: We prospectively evaluated 343 consecutive men with non-suspicious digital rectal examination and transrectal ultrasound findings and with serum PSA in the intermediate range (4.1-10 ng/mL). All men underwent systematic sextant biopsies of the peripheral zone. We performed a two fold analysis of PSAD performance first using measured gland volume and then using modified gland volumes ranging from a 25% underestimation to a 25% overestimation, in 5% stepwise increments. RESULTS: With a 0.15 PSAD cut-off, we would have missed 14, 34 and 48% of cancers, if we had respectively used a volume underestimate of 25%, the measured volume and a volume overestimate of 25%. CONCLUSIONS: Potential gland volume under- or overestimation may substantially affect the diagnostic performance of PSAD. Although PSAD may represent a useful adjunct in the early detection of prostate cancer, it may compromise cancer detection in a substantial proportion of cases, if used as the only indicator for biopsy. PMID- 12741974 TI - An assessment of CME needs of Canadian urologists. AB - The Canadian Urological Association has recently developed a new continuing medical education vehicle, Video Journal in Urology. The editorial content of this quarterly production is determined by the results of a needs assessment, the results of which are described in this paper. The needs assessment, as a two-page questionnaire, was mailed to 504 urologists. One hundred and eight-three responded, a response rate of 36.3%. The topic areas ranked as most interesting were largely centered around the broad topic of prostate cancer. PMID- 12741975 TI - Inferior vena cavectomy for renal cell carcinoma with intracaval tumor thrombus. AB - Excision of the inferior vena cava for renal cell carcinoma with intracaval tumor thrombus is infrequently performed. Herein the authors report a 60-year old woman with a right renal cell carcinoma and massive occluding tumor thrombus of the inferior vena cava. Following a negative metastatic workup, this patient underwent surgery to remove the tumor and thrombus. Thrombectomy occurred via excision of the affected portion of inferior vena cava and proximal left renal vein. Reconstruction of the vena cava was not undertaken. The patient did not suffer any morbidity during recovery in hospital. Her renal function was normal upon discharge. All resection margins were negative for tumor. This experience is compared to those reported in the literature. Postoperative morbidity may be minimized by careful patient selection. Suitable patients should have a right sided tumor with an occlusive subhepatic vena caval tumor thrombus. PMID- 12741977 TI - EDITORIAL. PMID- 12741976 TI - Prognostic value of angiogenesis and vascular invasion in muscle-invasive bladder cancer. AB - Angiogenesis is important in the pathogenesis of tumors and correlates with clinical outcome in certain malignancies. Recent advances in staining techniques have raised doubts about the prognostic value of vascular invasion in muscle invasive bladder cancer, for which relatively few predictors for survival exist. Twenty-one radical cystectomy specimens from patients with stage pT2-3NOMO transitional cell tumors were examined by staining with Hematoxylin and Eosin (H&E). Factor VIII-related antigen (FVIII RAG) which is specific for vascular endothelium, and MOVAT which highlights large vessels. Angiogenesis, estimated by microvessel density, and presence of vascular invasion were then correlated with survival in the follow-up period which ranged from 3 to 141 months. Six of 21 patients survived disease-free with follow-up ranging from 47 to 141 months. Twelve patients had lower microvessel density than 178/mm2 and five of these survived, whereas only one of nine patients having higher microvessel density survived. Twelve of 21 patients had vascular invasion and three survived whereas three of nine patients without vascular invasion survived. There was no correlation found between vascular invasion and survival in this study. Analysis by Kaplan-Meir survival curves demonstrated trends which were not statistically significant between high and low angiogenesis groups. The preliminary data suggest that further assessment of the prognostic role of angiogenesis and vascular invasion in muscle-invasive bladder cancer is indicated. PMID- 12741978 TI - Radiotherapy for localized prostate cancer: assessment of results by systematic biopsy and PSA. AB - This study attempted to determine the failure pattern after radiotherapy for localized prostate cancer using systematic biopsies and serum PSA in assessment of outcome. Between July 1987 and February 1993, 226 patients treated with radical external beam radiotherapy were followed prospectively with systematic transrectal ultrasound-guided biopsies and serum PSA. Four hundred and ten transrectal ultrasound-guided biopsies were performed with 4-7 samples (usually six) per session. Stage distribution was T1b: 32, T1c: 11, T2a: 45, T2b: 62, T3: 50, T4: 6. Median follow-up was 41 months. The rate of positive biopsies continued to decrease over time, reaching a nadir of 30.5% at 30 months post radiotherapy. Local failures have occurred in 17% (38/226) overall, or 13% of T1b, 0% of T1c, 13% of T2a, 20% of T2b and 21% of T3-4. Six additional patients (2.5%) have had biochemical failure and 34 (15%) have biopsy-only failure. Forty seven patients initially showing residual tumor on post radiotherapy biopsies (median time 18 months), demonstrated late tumor clearance, achieving negative biopsies at a median time of 28 months. Median nadir PSA in patients with no evidence of disease (NED) at last follow-up was 0.5 ng/mL, achieved at 24 months post radiotherapy. For patients with delayed tumor clearance, the median nadir PSA was 0.6 ng.mL at 27 months. For those with local failure it was 1.6 ng.mL at 16 months and for all failures, 1.8 ng/mL at 13 months. Systematic prostate biopsies show that tumor clearance after radiotherapy may take up to 30 months. Histologic tumor clearance parallels the fall in serum PSA. Serum PSA nadirs earlier and at a higher value in patients destined to fail. PMID- 12741979 TI - Tiaprofenic acid: an uncommon cause of cystitis often misdiagnosed as interstitial cystitis. AB - OBJECTIVE: To bring to the attention of Urologists that the medication tiaprofenic acid (Surgam, Albert-tiafen), a non-steroidal anti-inflammatory drug has been found to cause chemical cystitis. METHODS: Patients presenting between November 1994 and November 1995 with a confirmed or presumed diagnosis of interstitial cystitis and who were taking tiaprofenic acid are reported. RESULTS: Thirty-six patients presented with a confirmed or presumed diagnosis of interstitial cystitis. One woman without irritative symptoms presented with an abnormal ultrasound finding of a thickened bladder. Eight of these 37 patients were diagnosed with chemical cystitis from tiaprofenic acid. These eight patients (one man and seven women) with ages ranging between 46 and 79 (mean 67 years), were taking tiaprofenic acid from 22 to 84 months (mean 41.8 months), for the treatment of osteoarthritis. All eight patients had findings of inflammation at cystoscopy. Symptoms resolved from two to eight weeks (mean 5.3 weeks) in six out of the eight patients (one patient was asymptomatic and one is awaiting resolution two weeks after discontinuation), and persistence of bladder inflammation (as seen on cystoscopy) lingered greater than three months in two patients and five months one patient, after the medication was stopped. CONCLUSION: Symptoms of frequency, nocturia and suprapubic pain are associated with long term ingestion of tiaprofenic acid. Simple discontinuation of the drug will relieve symptoms usually within six weeks. The mechanism of inflammation is unknown. PMID- 12741981 TI - EDITORIAL. PMID- 12741980 TI - Urinary bladder endocervicosis: a case report and review of the literature. AB - Endocervicosis, a mucinous analogue of endometriosis, is a rare, non-neoplastic mullerian lesion that may involve the urinary bladder. This lesion has only recently been described in the pathologic literature and, to the best of our knowledge, it has not been previously reported in the urologic literature. It is a benign lesion that clinically may be confused with a malignant tumor. Histologically, it must be differentiated from an adenocarcinoma. We report one recent case of urinary bladder endocervicosis in a 34-year old female who presented with urinary urgency and frequency secondary to a 3.5 cm mass in the posterior wall of the bladder. This was treated by partial cystectomy with complete resolution of her symptoms. The literature on endocervicosis and related mullerian lesions of the urinary bladder is reviewed. PMID- 12741982 TI - Actin's actions in LTP-induced synapse growth. AB - F-actin is required for LTP induction, but the underlying mechanisms are unclear. New results show that LTP produces an increase in actin-rich spines. Furthermore, these spines have large synapses, strongly suggesting that LTP involves synapse growth. This process appears to require LIMK-1 phosphorylation of the actin depolymerizing factor cofilin. PMID- 12741984 TI - Seeing and acting at the same time: challenges for brain (and) research. AB - Traditionally, studies of the visual system of nonhuman primates have investigated neurons while the animal fixates a target in a static environment. Clearly, this is not what our everyday life is like; neither the environment nor we are stationary while we act in our world. On the contrary, we constantly move our eyes or limbs. Here we review a number of recent studies describing the brain in a more realistic mode of operation. PMID- 12741983 TI - Excitement reduces inhibition via endocannabinoids. AB - Previous work has suggested a role for retrograde synaptic signaling via endogenous cannabinoids in regulating the inhibitory control of neuronal activity. In this issue of Neuron, Chevaleyre and Castillo provide evidence for another form of endocannabinoid-mediated depression of hippocampal inhibition, which is activity dependent and long lasting. PMID- 12741985 TI - Progress and directions in olfactory development. PMID- 12741986 TI - Serine 776 of ataxin-1 is critical for polyglutamine-induced disease in SCA1 transgenic mice. AB - Polyglutamine-induced neurodegeneration in transgenic mice carrying the spinocerebellar ataxia type 1 (SCA1) gene is modulated by subcellular distribution of ataxin-1 and by components of the protein folding/degradation machinery. Since phosphorylation is a prominent mechanism by which these processes are regulated, we examined phosphorylation of ataxin-1 and found that serine 776 (S776) was phosphorylated. Residue 776 appeared to affect cellular deposition of ataxin-1[82Q] in that ataxin-1[82Q]-A776 failed to form nuclear inclusions in tissue culture cells. The importance of S776 for polyglutamine induced pathogenesis was examined by generating ataxin-1[82Q]-A776 transgenic mice. These mice expressed ataxin-1[82Q]-A776 within Purkinje cell nuclei, yet the ability of ataxin-1[82Q]-A776 to induce disease was substantially reduced. These studies demonstrate that polyglutamine tract expansion and localization of ataxin-1 to the nucleus of Purkinje cells are not sufficient to induce disease. We suggest that S776 of ataxin-1 also has a critical role in SCA1 pathogenesis. PMID- 12741987 TI - A role for BMP heterodimers in roof plate-mediated repulsion of commissural axons. AB - During spinal cord development, commissural neurons extend their axons ventrally, away from the roof plate. The roof plate is the source of a diffusible repellent that orients commissural axons in vitro and, thus, may regulate the trajectory of commissural axons in vivo. Of three Bmps expressed in the roof plate, BMP7, but not BMP6 or GDF7, mimics the roof plate activity in vitro. We show here that expression of both Bmp7 and Gdf7 by roof plate cells is required for the fidelity of commissural axon growth in vivo. We also demonstrate that BMP7 and GDF7 heterodimerize in vitro and that, under these conditions, GDF7 enhances the axon orienting activity of BMP7. Our findings suggest that a GDF7:BMP7 heterodimer functions as a roof plate-derived repellent that establishes the initial ventral trajectory of commissural axons. PMID- 12741988 TI - Peripheral NT3 signaling is required for ETS protein expression and central patterning of proprioceptive sensory afferents. AB - To study the role of NT3 in directing axonal projections of proprioceptive dorsal root ganglion (DRG) neurons, NT3(-/-) mice were crossed with mice carrying a targeted deletion of the proapoptotic gene Bax. In Bax(-/-)/NT3(-/-) mice, NT3 dependent neurons survived and expressed the proprioceptive neuronal marker parvalbumin. Initial extension and collateralization of proprioceptive axons into the spinal cord occurred normally, but proprioceptive axons extended only as far as the intermediate spinal cord. This projection defect is similar to the defect in mice lacking the ETS transcription factor ER81. Few if any DRG neurons from Bax(-/-)/NT3(-/-) mice expressed ER81 protein. Expression of a NT3 transgene in muscle restored DRG ER81 expression in NT3(-/-) mice. Finally, addition of NT3 to DRG explant cultures resulted in induction of ER81 protein. Our data indicate that NT3 mediates the formation of proprioceptive afferent-motor neuron connections via regulation of ER81. PMID- 12741989 TI - ATP-sensitive potassium channel traffic regulation by adenosine and protein kinase C. AB - ATP-sensitive potassium (K(ATP)) channels activate under metabolic stress to protect neurons and cardiac myocytes. However, excessive channel activation may cause arrhythmia in the heart and silence neurons in the brain. Here, we report that PKC-mediated downregulation of K(ATP) channel number, via dynamin-dependent channel internalization, can act as a brake mechanism to control K(ATP) activation. A dileucine motif in the pore-lining Kir6.2 subunit of K(ATP), but not the site of PKC phosphorylation for channel activation, is essential for PKC downregulation. Whereas K(ATP) activation results in a rapid shortening of the action potential duration (APD) in metabolically inhibited ventricular myocytes, adenosine receptor stimulation and consequent PKC-mediated K(ATP) channel internalization can act as a brake to lessen this APD shortening. Likewise, in hippocampal CA1 neurons under metabolic stress, PKC-mediated, dynamin-dependent K(ATP) channel internalization can also act as a brake to dampen the rapid decline of excitability due to K(ATP) activation. PMID- 12741990 TI - Shunting inhibition modulates neuronal gain during synaptic excitation. AB - Neuronal gain control is important for processing information in the brain. Shunting inhibition is not thought to control gain since it shifts input-output relationships during tonic excitation rather than changing their slope. Here we show that tonic inhibition reduces the gain and shifts the offset of cerebellar granule cell input-output relationships during frequency-dependent excitation with synaptic conductance waveforms. Shunting inhibition scales subthreshold voltage, increasing the excitation frequency required to attain a particular firing rate. This reduces gain because frequency-dependent increases in input variability, which couple mean subthreshold voltage to firing rate, boost voltage fluctuations during inhibition. Moreover, synaptic time course and the number of inputs also influence gain changes by setting excitation variability. Our results suggest that shunting inhibition can multiplicatively scale rate-coded information in neurons with high-variability synaptic inputs. PMID- 12741991 TI - Hippocampal LTP is accompanied by enhanced F-actin content within the dendritic spine that is essential for late LTP maintenance in vivo. AB - The dendritic spine is an important site of neuronal plasticity and contains extremely high levels of cytoskeletal actin. However, the dynamics of the actin cytoskeleton during synaptic plasticity and its in vivo function remain unclear. Here we used an in vivo dentate gyrus LTP model to show that LTP induction is associated with actin cytoskeletal reorganization characterized by a long-lasting increase in F-actin content within dendritic spines. This increase in F-actin content is dependent on NMDA receptor activation and involves the inactivation of actin depolymerizing factor/cofilin. Inhibition of actin polymerization with latrunculin A impaired late phase of LTP without affecting the initial amplitude and early maintenance of LTP. These observations suggest that mechanisms regulating the spine actin cytoskeleton contribute to the persistence of LTP. PMID- 12741992 TI - Heterosynaptic LTD of hippocampal GABAergic synapses: a novel role of endocannabinoids in regulating excitability. AB - Neuronal excitability and long-term synaptic plasticity at excitatory synapses are critically dependent on the level of inhibition, and accordingly, changes of inhibitory synaptic efficacy should have great impact on neuronal function and neural network processing. We describe here a form of activity-dependent long term depression at hippocampal inhibitory synapses that is triggered postsynaptically via glutamate receptor activation but is expressed presynaptically. That is, glutamate released by repetitive activation of Schaffer collaterals activates group I metabotropic glutamate receptors at CA1 pyramidal cells, triggering a persistent reduction of GABA release that is mediated by endocannabinoids. This heterosynaptic form of plasticity is involved in changes of pyramidal cell excitability associated with long-term potentiation at excitatory synapses and could account for the effects of cannabinoids on learning and memory. PMID- 12741993 TI - Robust spatial working memory through homeostatic synaptic scaling in heterogeneous cortical networks. AB - The concept of bell-shaped persistent neural activity represents a cornerstone of the theory for the internal representation of analog quantities, such as spatial location or head direction. Previous models, however, relied on the unrealistic assumption of network homogeneity. We investigate this issue in a network model where fine tuning of parameters is destroyed by heterogeneities in cellular and synaptic properties. Heterogeneities result in the loss of stored spatial information in a few seconds. Accurate encoding is recovered when a homeostatic mechanism scales the excitatory synapses to each cell to compensate for the heterogeneity in cellular excitability and synaptic inputs. Moreover, the more realistic model produces a wide diversity of tuning curves, as commonly observed in recordings from prefrontal neurons. We conclude that recurrent attractor networks in conjunction with appropriate homeostatic mechanisms provide a robust, biologically plausible theoretical framework for understanding the neural circuit basis of spatial working memory. PMID- 12741995 TI - Hemispheric dissociation in access to the human semantic system. AB - Patient studies suggest that speech and environmental sounds are differentially processed by the left and right hemispheres. Here, using functional imaging in normal subjects, we compared semantic processing of spoken words to equivalent processing of environmental sounds, after controlling for low-level perceptual differences. Words enhanced activation in left anterior and posterior superior temporal regions, while environmental sounds enhanced activation in a right posterior superior temporal region. This left/right dissociation was unchanged by different attentional/working memory contexts, but it was specific to tasks requiring semantic analysis. While semantic processing involves widely distributed networks in both hemispheres, our results support the hypothesis of a dual access route specific for verbal and nonverbal material, respectively. PMID- 12741996 TI - Neural manifestations of memory with and without awareness. AB - Neurophysiological events responsible for different types of human memory tend to occur concurrently and are therefore difficult to measure independently. To surmount this problem, we produced perceptual priming (indicated by speeded responses) in the absence of conscious remembering. At encoding, faces appeared briefly while subjects' attention was diverted to other stimuli. Faces appeared again in either an implicit or explicit memory test. Neural correlates of priming were identified as brain potentials beginning 270 ms after face onset with more negative amplitudes for repeated than for new faces. Remembered faces, in contrast, activated a different configuration of intracranial sources producing positive potentials maximal at 600-700 ms. We thus disentangled and characterized distinct neural events associated with memory with and without awareness. PMID- 12741994 TI - Spatiotemporal dynamics of modality-specific and supramodal word processing. AB - The ability of written and spoken words to access the same semantic meaning provides a test case for the multimodal convergence of information from sensory to associative areas. Using anatomically constrained magnetoencephalography (aMEG), the present study investigated the stages of word comprehension in real time in the auditory and visual modalities, as subjects participated in a semantic judgment task. Activity spread from the primary sensory areas along the respective ventral processing streams and converged in anterior temporal and inferior prefrontal regions, primarily on the left at around 400 ms. Comparison of response patterns during repetition priming between the two modalities suggest that they are initiated by modality-specific memory systems, but that they are eventually elaborated mainly in supramodal areas. PMID- 12741997 TI - Mitochondrial K ATP channel activation is important in the antiarrhythmic and cardioprotective effects of non-hypotensive doses of nicorandil and cromakalim during ischemia/reperfusion: a study in an intact anesthetized rabbit model. AB - The roles of cardiomyocyte sarcolemmal ATP-sensitive K(+) (K(ATP)) and mitochondrial K(ATP) channels in the cardioprotection and antiarrhythmic activity induced by K(ATP) channel openers remain obscure, though the mitochondrial K(ATP) channels have been proposed to be involved as a subcellular mediator in cardioprotection afforded by ischemic preconditioning. In the present study, we investigated the effects of administration of non-hypotensive doses of ATP sensitive K(+) channel (K(ATP)) openers (nicorandil and cromakalim), a specific mitochondrial K(ATP) channel blocker (5-hydroxydecanoate (5-HD)) and a specific sarcolemmal K(ATP) channel blocker (HMR 1883; (1-[5-[2-(5-chloro-o anisamido)ethyl]-2-methoxyphenyl]sulfonyl-3-methylthiourea) prior to and during coronary occlusion as well as prior to and during post-ischemic reperfusion on survival rate, ischemia-induced and reperfusion-induced arrhythmias and myocardial infarct size in anesthetized albino rabbits. The thorax was opened in the left 4th intercostal space and after pericardiotomy the heart was exposed. In Group I (n=80), occlusion of the left main coronary artery and hence, myocardial ischemia-induced arrhythmias were achieved by tightening a previously placed loose silk ligature for 30min. In Group II (n=184), arrhythmias were induced by reperfusion following a 20 min ligation of the left main coronary artery. Both in Groups I and II, early intravenous infusion of nicorandil (100 micro g/kg bolus+10 micro g/kg/min), cromakalim (0.2 micro g/kg/min), HMR 1883 (3mg/kg)/nicorandil and HMR 1883 (3mg/kg)/cromakalim just prior to and during ischemia increased survival rate (75%, 67%, 86% and 75% versus 60% in the control subgroup in Group I; 75%, 75%, 75% and 67% versus 50% in the control subgroup in Group II), significantly decreased the incidence and severity of life-threatening arrhythmias and significantly decreased myocardial infarct size. However, late intravenous administration of nicorandil or cromakalim at the onset and during reperfusion did neither increase survival rate nor confer any antiarrhythmic or cardioprotective effects. The antiarrhythmic and cardioprotective effects of both nicorandil and cromakalim were abolished by pretreating the rabbits with 5-HD (5mg/kg, i.v. bolus), a selective mitochondrial K(ATP) channel blocker but not by HMR 1883 (3mg/kg). In the present study, higher levels of malondialdehyde (MDA) and lower levels of reduced glutathione (GSH) and superoxide dismutase (SOD) in necrotic zone of myocardium in all the 16 subgroups in Group II suggest little anti-free radical property of nicorandil and cromakalim.We, therefore, conclude that intervention by intravenous administration of nicorandil and cromakalim (through the selective activation of mitochondrial K(ATP) channels), increased survival rate and exhibited antiarrhythmic and cardioprotective effects during coronary occlusion and reperfusion in anesthetized rabbits when administered prior to and during coronary occlusion. The mitochondrial K(ATP) channel may be a potential site of cardioprotection and antiarrhythmic activity. PMID- 12741998 TI - Grape seed proanthocyanidin extract attenuates oxidant injury in cardiomyocytes. AB - This study sought to test whether grape seed proanthocyanidin extract (GSPE) attenuates exogenous and endogenous oxidant stress induced in chick cardiomyocytes and whether this cytoprotection is mediated by PKC activation, mito K(ATP) channel opening, NO production, oxidant scavenging, or iron chelating effects. Cells were exposed to hydrogen peroxide (H(2)O(2)) (exogenous oxidant stress, 0.5mM) or antimycin A (endogenous oxidant stress, 100 micro M) for 2h following pretreatment with GSPE at various concentrations for 2h. Cells were also pretreated with GSPE or with inhibitors of PKC (chelerytherine), mito K(ATP) channel (5-hydroxydecanoate), nitric oxide synthase (nitro-L-arginine methyl ester) for 2h. Oxidant stress was measured by 2',7'-dichlorofluorescin diacetate and cell viability was assessed using propidium iodide. Free radical scavenging and iron chelating ability was tested in vitro. GSPE dose-dependently attenuated oxidant formation and significantly improved cell survival and contractile function. However, inhibitors of PKC, mito K(ATP) channel or NO synthase failed to abolish the protective action of GSPE during H(2)O(2) or antimycin A exposure. In vitro studies suggested that GSPE scavenges H(2)O(2), hydroxyl radical and superoxide, and may chelate iron. These results indicate that GSPE confers cardioprotection against exogenous H(2)O(2)- or antimycin A-induced oxidant injury. Its effect does not require PKC, mito K(ATP) channel, or NO synthase, presumably because it acts by reactive oxygen species scavenging and iron chelating directly. PMID- 12741999 TI - Anti-inflammatory activity of 3-benzoyl-1-methyl-4-phenyl-4-piperidinol hydrochloride. AB - In our study, the effects of the compound 3-benzoyl-1-methyl-4-phenyl-4 piperidinol hydrochloride (C1) and also an anti-inflammatory drug, indomethacin, tested by carrageenan-induced paw edema and cotton pellet granuloma tests, for their effects on acute and chronic phases of inflammation, respectively. Their effects on vascular permeability were also tested by hyaluronidase-induced capillary permeability. C1 decreased the carrageenan-induced paw edema 61.98, 80.84, and 90.32% at 50, 100, and 200mgkg(-1) doses, respectively, while this decrease was 89.93% by indomethacin at 20mgkg(-1) dose. Antiproliferative effects of C1 at 100mgkg(-1) and indomethacinin at 20mgkg(-1) doses were 46.1 and 43.1%, respectively, in cotton pellet test. C1 and indomethacin both significantly inhibited the hyaluronidase-induced increase in capillary permeability. PMID- 12742000 TI - Effect of vigabatrin on contractile response to arachidonic acid and prostaglandins in smooth muscle preparations and platelet aggregation in experimental laboratory animals. AB - Gamma-vinyl GABA (vigabatrin; VGB), an irreversible inhibitor of GABA transaminase, was evaluated for its effect on the contractile responses to PGE(2) or its precursor arachidonic acid (AA) in guinea pig ileum (GPI) and non-pregnant rat uterus. Rationale behind this study was the rise in gamma-aminobutyric acid (GABA) contents in the peripheral organs after treatment with GABAergic agents and extensive interconnections of neuromediators and their interactions.Indomethacin, the standard NSAID, at 6.1x10(-5) and 12.2x10(-5)M concentrations highly significantly (P<0.001) inhibited contractile responses induced by AA (4.3x10(-5)M) in the isolated GPI. Incubation of the GPI segments with VGB at different concentrations (25, 100 and 200mM) failed to inhibit AA induced contractile responses in the same preparation. In fact, VGB at 25mM significantly (P<0.05) potentiated the contractile effect of AA 4.3x10(-5)M. Incubation of the same tissue with GABA at 13x10(-3) and 26x10(-3)M inhibited responses elicited by AA at 4.3x10(-5)M that was significant (P<0.05) only with low concentration of GABA. Conversely, a higher concentration (64x10(-3)M) of GABA was needed to antagonise PGE(2) (9.4x10(-6)M)-induced contractions in the same tissue. Almost similar results (i.e. inhibition of contractile responses to AA and PGE(2)) were obtained with isolated non-pregnant rat uterus. These findings suggested that GABA had the potential to inhibit prostaglandin (PG) synthesis and/or antagonise PGE(2) responses in isolated GPI and rat uterus. The addition of 35 or 70 micro l of rat aorta incubation medium caused a dose dependent inhibition of ADP-induced aggregation being 21.5 and 43.5%, respectively, an indication of prostacycline (PGI(2)) contents. Pre-incubation of rat aortic tissues with VGB (96.8mM) significantly (P<0.05) reversed the anti aggregatory activity of control aortic prostacycline on ADP-induced aggregation. These findings suggested that VGB might inhibit PGI(2) activity through the inhibition of its synthesis. PMID- 12742001 TI - Effect of a selective inducible nitric oxide synthase inhibitor on intraocular nitric oxide production in endotoxin-induced uveitis rabbits: in vivo intraocular microdialysis study. AB - Inducible nitric oxide (NO) synthase (iNOS) is believed to contribute to the pathogenesis of endotoxin-induced uveitis (EIU). In the present study, we investigated the inhibitory effects of N(G)-nitro-L-arginine methyl ester (L NAME), a non-selective NOS inhibitor, and S,S'-1,4-phenylene-bis(1,2 ethanediyl)bis-isothiourea (PBITU), a potent and selective iNOS inhibitor, on intraocular NO production in EIU rabbits using an in vivo intraocular microdialysis technique. The flare level in the anterior chamber increased from 1h after the injection of 100 micro g/kg lipopolysaccharide (LPS), and continued to increase for 24h. Aqueous humor protein concentrations were significantly increased at 24h after LPS-injection. These changes were significantly reduced by L-NAME (10mg/kg) and PBITU (1mg/kg), but not by D-NAME (10mg/kg). The increase in NO(2)(-) and NO(3)(-) levels in the dialysate induced by LPS was significantly inhibited by L-NAME (10mg/kg) and PBITU (1mg/kg), but not by D-NAME (10mg/kg). These results suggest that activation of iNOS may play a key role in the development of EIU, and selective inhibitors of iNOS may have therapeutic applications in the treatment of EIU. PMID- 12742002 TI - Retrospective analysis of adverse drug reactions to bronchodilators observed in two pulmonary divisions of Catanzaro, Italy. AB - We retrospectively analysed the adverse drug reactions (ADRs) associated with bronchodilator therapy and reported over a 7-year period, from January 1995 to December 2001, in clinical notes of two Pulmonary division of "Mater Domini" University Hospital and "Pugliese-Ciaccio" Hospital, both located in Catanzaro, Italy. Bronchodilators were responsible for 45 (18.5%) out of 243 episodes of ADRs. Theophylline was the drug most involved in ADRs (53.4%), and skin was the body system most susceptible to ADRs induced by all bronchodilators (47.7%). We determined that the drug-ADR relationship was certain in 73% of the reports; withdrawal of the suspected drug led to recovery in 86% of cases. In conclusion, this retrospective evaluation demonstrated that bronchodilators are a common cause of ADRs in hospitalised patients and, therefore, drug surveillance can successfully identify adverse events related with drug administration in hospitalised patients. PMID- 12742003 TI - Drugs and non-medical products sold in pharmacy: information and advertising. AB - This paper is the third part of a survey about patient information and the use of patient package leaflet. The aim of this research section is to analyse the role of the pharmacy and the pharmacist in the health-care field. By means of answers of interviewed people, informations were obtained about the purchase of drugs and other products in pharmacy, the consumer requirements, the non-medical products and self-medication channels of information, the patient-physician and patient pharmacist quality of communication and the role of advertising for drugs and other products sold in pharmacy. From this survey, it emerged that most of the respondents go to pharmacy to purchase drugs exclusively and only a small number of people go to purchase other products. The pharmacist does not play a remarkable role in providing health information to the patient. PMID- 12742004 TI - Testicular toxicity of dibromoacetonitrile and possible protection by tertiary butylhydroquinone. AB - Dibromoacetonitrile (DBAN) is known to be water disinfectant by-product. Its broad-spectrum toxicity in different test systems in vivo and in vitro has been reported. Oxidative damage induced by DBAN may be partially responsible for its toxicity. Herein, the ability of DBAN to induce oxidative stress in mouse testis and possible protective effect of an antioxidant tertiary butylhydroquinone (TBHQ) were addressed. Male albino mice were injected with a single dose of DBAN (50mg/kg i.p.), and killed after 3h of treatment. Control animals received 10ml/kg body weight i.p. of the vehicle DMSO. In both experiments, cauda epididymis were dissected and sperm count and motility were investigated. Also, testicular activity of lactic dehydrogenase-x (LDH-x) isozyme and histopathological changes were examined. Furthermore, testicular content of malonyldialdehyde (MDA) and reduced glutathione (GSH) were determined. A single i.p. dose of DBAN caused decrease in sperm count and motility to approximately 88 and 84%, respectively, compared with control animals. A 46% decrease in testicular activity of LDH-x, compared with control animals, was observed. A significant accumulation of MDA in DBAN-treated animals was increased to 99% while testicular content of GSH was decreased by 56% compared to control animals. Compared to DBAN-treated animals, treatment with TBHQ (100mg/kg p.o.) prior exposure to DBAN showed a remarkable degree of protection as indicated by enhancement of sperm count and motility, testicular activity of LDH-x, and GSH. Accumulation of testicular content of MDA significantly decreased following TBHQ treatment compared to DBAN-treated animals. In conclusion, results presented here indicate that DBAN is capable to induce oxidative stress in mouse testis. TBHQ may play a protective role against DBAN-induced testicular cellular damage. PMID- 12742005 TI - The effects of oral glutamine on cisplatin-induced nephrotoxicity in rats. AB - The antioxidant activity of the amino acid glutamine was investigated to obtain protection against peroxidative damage in rat kidney and nephrotoxicity induced by the treatment with a single dose of the antitumoral cisplatin (5mgkg(-1) body weight). The animals were divided into four treatment and control groups of six rats each (n=6). Cisplatin was injected i.p. and glutamine (300mgkg(-1) body weight) was given by gavage 24h before the cisplatin injection. After 24h and 7 days of cisplatin administration, the rats were sacrificed. A single dose of cisplatin resulted in significant reduction in body weight and creatinine clearance, and higher urinary volumes were observed in all groups treated with this antitumor drug (P<0.05). Renal tissue from cisplatin-treated rats showed an increase in malondialdehyde production and increase in glutathione contents 24h and 7 days after cisplatin administration. Pretreatment of rats with glutamine substantially inhibited the increase in the levels of renal glutathione induced by cisplatin 24h after the i.p. injection. The malondialdehyde in the renal tissues was significantly reduced 7 days after cisplatin treatment. However, the reduction in the peroxidative damage did not reach the value of the control group. The protective effects obtained by glutamine pretreatment in peroxidative alterations were not observed in the other parameters studied. These results suggest that glutamine partially protect against cisplatin-induced lipid peroxidation damage, but it was not enough to inhibit cisplatin-induced nephrotoxicity in rats. PMID- 12742006 TI - A possible mechanism of gastrointestinal toxicity posed by mycophenolic acid. PMID- 12742007 TI - The effect of combined administration of ethanol and sertraline, fluoxetine and citalopram on rabbit EEG. AB - In this study we have decided to examine acute interaction of ethanol with some drugs that belong to selective serotonin inhibitor (SSRI) group. Therefore, the influence of sertraline, fluoxetine and citalopram on the effect of ethanol on EEG of rabbits (frontal cortex, hippocampus, MRF) was tested. Sertraline (10mg/kg i.p.), fluoxetine (10mg/kg i.p.) and citalopram (5mg/kg i.p.) were given 30min before ethanol injection in a dose 0.8g/kg i.v. Ethanol caused the increase of the slow frequencies (0.5-4cps) in the recording, as well as a marked decrease of the fastest frequencies (13-30 and 30-45cps). Sertraline, fluoxetine and citalopram (given before ethanol) prevented the increase in the slow frequencies (0.5-4cps) in the recordings from the frontal cortex and hippocampus, which indicates on antagonism inhibitory action of ethanol. These drugs administered together with ethanol may increase its influence on fast frequencies. This effect depends on brain structure and drug. PMID- 12742008 TI - Effects of ranitidine on pulmonary function tests of patients with chronic obstructive pulmonary disease. AB - Since the incidence of peptic ulcer and gastroesophageal reflux (GER) is more common in patients with chronic obstructive pulmonary disease (COPD) than normal population, H(2) receptor blockers are given more extensively to COPD patients. This study evaluated the effects of Ranitidine on pulmonary function tests (PFT) of the patients having COPD and peptic ulcer or GER, and of healthy volunteers. Fifty milligrams of Ranitidine was given intravenously to 30 COPD patients and 25 healthy volunteers. PFT were done before and 15, 30, 60, 120min after Ranitidine injection. Although mean forced vital capacity (FVC), forced expiratory volume in 1s (FEV(1)) and forced midexpiratory flow rate (FEF(25-75%)) of COPD patients were found to be decreased 60 and 120min after Ranitidine injection, the decrements were statistically insignificant. The decrements in PFT of healthy volunteers were also not statistically significant.H(2) receptor blockers can be used safely for treatment of gastrointestinal disorders in COPD patients who have mild or moderate obstruction. Minimal decreases in FEV(1) and FVC due to treatment by H(2) receptor blockers may clinically worsen COPD patients who have severe obstruction. PMID- 12742009 TI - Effects of zafirlukast on bronchial asthma and allergic rhinitis. AB - Asthma and allergic rhinitis are common conditions, occurring with increasing prevalence and frequently coexist. In both conditions histamine and cysteinil leukotrienes are important pathogenic inflammatory mediators. We evaluated the effects of the leukotriene receptor antagonist zafirlukast, 20mg administered twice daily for 2 weeks, in patients with allergic rhinitis and bronchial asthma during the grass pollen season. Patients underwent skin prick testing, spirometry, rhinomanometry, mucus transport test with saccharine, nasal epithelial brushing to study ciliary beat and, finally, nasal lavage.Thirty-five subjects completed the study. At the end of the study period, zafirlukast significantly reduced asthma and rhinitis symptoms (P< or =0.05); FEV(1) values were unchanged (P=0.10), whereas nasal resistances showed a decrease following treatment (P=0.01). Ciliary beat frequency (CBF) also improved (P=0.00), although mucociliary transport showed no improvement (P=0.87). The number of eosinophils in nasal lavage fluid decreased (P=0.00) while that of neutrophils was unchanged (P=0.09). These positive effects suggest that zafirlukast may be usefully employed in the treatment of both bronchial asthma, as previously demonstrated, and allergic rhinitis. PMID- 12742010 TI - Influence of a 3-day regimen of azithromycin on the disposition kinetics of cyclosporine A in stable renal transplant patients. AB - Some macrolide antibiotics have been shown to produce significant drug-drug interactions through the inhibition of cytochrome P450 (CYP) enzymes. In renal transplant patients these interactions pose potentially serious problems for the safe administration of cyclosporine A (CSA), a substrate of CYP3A4. The effects of azithromycin on CSA disposition kinetics were evaluated in eight stable renal transplant patients. Patients had been stabilized on individualized doses of CSA which remained unchanged throughout the study. Azithromycin was administered for 3 days. Baseline measurements of CSA disposition kinetics were taken prior to azithromycin treatment (study day 2) and after 3 days (study day 5) of azithromycin treatment (500mg/day, orally). The key parameters of interest were the area under the CSA blood concentration versus time curve (AUC) measured for 24h after the morning dose of CSA on both days 2 and 5, and the C(max) values of CSA. The geometric mean ratios (GMRs) of those parameters (day 5/day 2) and their 90% confidence intervals (90% CI) were 107 (98,116) and 119 (104,136), respectively. The 7% increase in exposure level and 19% increase in peak plasma concentration are not likely to be clinically significant. It is concluded that azithromycin (500mg/dayx3 days) does not alter the disposition kinetics of CSA in a clinically significant way, and that CSA dosage adjustments are not warranted in renal transplant patients taking these two drugs together. PMID- 12742011 TI - Poloxamer gel as vehicle for transdermal iontophoretic delivery of arginine vasopressin: evaluation of in vivo performance in rats. AB - The present study describes the formulation and evaluation for pharmacokinetic and pharmacodynamic activity of arginine vasopressin (AVP), a nanopeptide with antidiuretic activity on being delivered by transdermal iontophoresis. Poloxamer 407 was used to form stable gels that did not reduce the release of AVP. The release rate from the gel followed Higuchi kinetics indicating that the dominant mechanism of release is diffusion. Iontophoresis alone and in combination with chemical enhancers was used to augment the transdermal permeation of AVP. The results of both pharmacokinetic and pharmacodynamic studies emphasize the dimension of 'rapid onset' achieved by iontophoresis. The correlation between pharmacokinetic data and pharmacodynamic activity was only qualitative. Histopathological studies revealed that skin toxicity caused by either iontophoresis or chemical enhancers when used alone could be reduced by using a combination of both the techniques in tandem. PMID- 12742012 TI - Effect of iontophoresis and fatty acids on permeation of arginine vasopressin through rat skin. AB - The aim of this study was to assess the effects of fatty acids and iontophoretic mode of penetration enhancement on transdermal delivery of Arginine Vasopressin (AVP). Sprague-Dawley (SD) rat skin was pretreated with fatty acids (e.g. 5% w/v, lauric acid, oleic acid, and linoleic acid in ethanol:water (EtOH:W, 2:1 system) for 2h and iontophoresis in vitro, separately or together. The results indicate that all fatty acids studied increased (P<0.05) the flux of AVP in comparison to control (not pretreated with enhancer) and their effectiveness in flux enhancement was comparable. Further, oleic acid in combination with iontophoresis significantly increased the permeation of AVP both in comparison to pretreatment with fatty acids and iontophoresis alone. However, iontophoresis did not further increase the permeation of AVP through linoleic acid pretreated skin. Fourier transform infrared (FT-IR) spectroscopic studies revealed that EtOH:W (2:1) system is not effective in lipid extraction. The shift to higher wavenumbers of the symmetric and asymmetric stretching peaks at 2850 and 2920cm(-1) revealed that at the concentration used, oleic acid and linoleic acid caused fluidization of stratum corneum (SC) lipids. This study provides direct evidence that oleic acid in EtOH:W (2:1) system causes disruption of the SC lipid lamellae and that a combination of oleic acid with iontophoresis further enhances the effects of oleic acid in a synergistic manner. PMID- 12742013 TI - Quantitative account of the enhanced affinity of two linked scFvs specific for different epitopes on the same antigen. AB - Protein and other antigens typically have a number of different epitopes. This presents an opportunity for designing high-affinity antibodies by connecting via a flexible peptide linker two antibody fragments recognizing non-overlapping epitopes on the same antigen. The same strategy was employed in natural and designed DNA-binding proteins. According to a previous theory, the linking enhances the antigen-binding affinity over those of the individual antibody fragments (with association constants K(A) and K(B)) by p(d(0))K(B) or p(d(0))K(A), where p(d(0))=(3/4pil(p)bL)(3/2)exp(-3d(0)(2)/4l(p)bL)(1-5l(p)/4bL+ cdots, three dots, centered ) is the probability density for the end-to-end vector of the flexible linker with L residues to have a distance d(0). The predicted affinity enhancement is found to be actually approached by a bi specific antibody against hen egg lysozyme consisting of scFv fragments of D1.3 and HyHEL-10. The wide applicability of the theory is demonstrated by diverse examples of protein-protein interactions constrained by flexible linkers. PMID- 12742014 TI - Orientation of the 19S regulator relative to the 20S core proteasome: an immunoelectron microscopic study. AB - Specific labelling with monoclonal antibodies reveals that in regulator proteasome complexes the asymmetric 19S regulator (PA700) binds to one or both terminal alpha-disks of the cylinder-shaped 20S core proteasome in such a way that its reclining front part is positioned in the vicinity of proteasome subunit alpha6. The protruding rear part of the regulator appears to be situated distal to the sites occupied by the subunits alpha2 and alpha3, respectively. When viewed from beta1/beta1' to beta4/beta4' along the polar 2-fold axis of the 20S proteasome core, the rear part of each 19S regulator cap appears to protrude clockwise. Thus, a defined alignment of the 19S regulator with respect to the single polar 2-fold rotational axis of the 20S core proteasome is obtained. PMID- 12742015 TI - An atomic model for actin binding by the CH domains and spectrin-repeat modules of utrophin and dystrophin. AB - Utrophin and dystrophin link cytoskeletal F-actin filaments to the plasmalemma. Genetic strategies to replace defective dystrophin with utrophin in individuals with muscular dystrophy requires full characterization of these proteins. Both contain homologous N-terminal actin-binding motifs composed of a pair of calponin homology (CH) domains (CH1 and CH2) that are connected by spectrin-repeat modules to C-terminal membrane-binding sequences. Here, electron microscopy and 3D reconstruction of F-actin decorated with utrophin and dystrophin actin-binding constructs were performed using Utr261 (utrophin's CH domain pair), Utr416 (utrophin's CH domains and first spectrin-repeat) and Dys246 (dystrophin's CH domain pair). The lozenge-like utrophin CH domain densities localized to the upper surface of actin subdomain 1 and extended azimuthally over subdomain 2 toward subdomains 3 and 4. The cylinder-shaped spectrin-repeat was located at the end of the CH domain pair and was aligned longitudinally along the cleft between inner and outer actin domains, where tropomyosin is present when on thin filaments. The connection between the spectrin-repeat module and the CH domains defined the orientation of CH1 and CH2 on actin. Resolution of utrophin's CH domains and spectrin-repeats permitted docking of crystal structures into respective EM densities, leading to an atomic model where both CH and spectrin domains bind actin. The CH domain-actin interaction for dystrophin was found to be more complex than for utrophin. Binding assays showed that Utr261 and Utr416 interacted with F-actin as monomers, whereas Dys246 appeared to associate as a dimer, consistent with a bilobed Dys246 structure observed on F-actin in electron microscope reconstructions. One of the lobes was similar in shape, position and orientation to the monomeric CH domains of Utr261, while the other lobe apparently represented a second set of CH domains in the dimeric Dys246. The extensive contact made by dystrophin on actin may be used in vivo to help muscles dissipate mechanical stress from the contractile apparatus to the extracellular matrix. PMID- 12742016 TI - A mechanism for ParB-dependent waves of ParA, a protein related to DNA segregation during cell division in prokaryotes. AB - Prokaryotic plasmids encode partitioning (par) loci involved in segregation of DNA to daughter cells at cell division. A functional fusion protein consisting of Walker-type ParA ATPase and green fluorescent protein (Gfp) oscillates back and forth within nucleoid regions with a wave period of about 20 minutes. A model is discussed which is based on cooperative non-specific binding of ParA to the nucleoid, and local ParB initiated generation of ParA oligomer degradation products, which act autocatalytically on the degradation reaction. The model yields self-initiated spontaneous pattern formation, based on Turing's mechanism, and these patterns are destroyed by the degradation products, only to initiate a new pattern at the opposite nucleoid region. A recurrent wave thus emerges. This may be a particular example of a more general class of pattern forming mechanisms, based on protein oligomerization upon a template (membranes, DNA a.o.) with resulting enhanced NTPase function in the oligomer state, which may bring the oligomer into an unstable internal state. An effector initializes destabilization of the oligomer to yield degradation products, which act as seeds for further degradation in an autocatalytic process. We discuss this mechanism in relation to recent models for MinDE oscillations in E.coli and to microtubule degradation in mitosis. The study points to an ancestral role for the presented pattern types in generating bipolarity in prokaryotes and eukaryotes. PMID- 12742017 TI - Structural determinants in human DNA polymerase gamma account for mitochondrial toxicity from nucleoside analogs. AB - Although antiviral nucleoside analog therapy successfully delays progression of HIV infection to AIDS, these drugs cause unwelcome side-effects by inducing mitochondrial toxicity. We and others have demonstrated that the mitochondrial polymerase, DNA polymerase gamma (pol gamma), participates in mitochondrial toxicity by incorporating these chain-terminating antiviral nucleotide analogs into DNA. Here, we explore the role of three highly conserved amino acid residues in the active site of human pol gamma that modulate selection of nucleotide analogs as substrates for incorporation. Sequence alignments, crystal structures and mutagenesis studies of family A DNA polymerases led us to change Tyr951 and Tyr955 in polymerase motif B to Phe and Ala, and Glu895 in polymerase motif A was changed to Ala. The mutant polymerases were tested for their ability to incorporate natural nucleotides and the five antiviral nucleoside analogs currently approved for antiviral therapy: AZT, ddC, D4T, 3TC and carbovir. Steady state kinetic analysis of the pol gamma derivatives with the normal and antiviral nucleotides demonstrated that Tyr951 is largely responsible for the ability of pol gamma to incorporate dideoxynucleotides and D4T-MP. Mutation of Tyr951 to Phe renders the enzyme resistant to dideoxynucleotides and D4T-TP without compromising the activity of the polymerase. Alteration of Glu895 and Tyr955 to Ala had the largest effect on overall polymerase activity with normal nucleotides, producing dramatic increases in K(m(dNTP)) and large decreases in k(cat). Mutation of Tyr955 in pol gamma causes the degenerative disease progressive external ophthalmoplegia in humans, and we show that this residue partially accounts for the ability of pol gamma to incorporate D4T-MP and carbovir. Alteration of Glu895 to Ala slightly increased discrimination against dideoxynucleotides and D4T-TP. The mechanisms by which pol gamma selects certain nucleotide analogs are discussed. PMID- 12742018 TI - Interplay between DnaA and SeqA proteins during regulation of bacteriophage lambda pR promoter activity. AB - DnaA and SeqA proteins are main regulators (positive and negative, respectively) of the chromosome replication in Escherichia coli. Nevertheless, both these replication regulators were found recently to be also transcription factors. Interestingly, both DnaA and SeqA control activity of the bacteriophage lambdap(R) promoter by binding downstream of the transcription start site, which is unusual among prokaryotic systems. Here we asked what are functional relationships between these two transcription regulators at one promoter region. Both in vivo and in vitro studies revealed that DnaA and SeqA can activate the p(R) promoter independently and separately rather than in co-operation, however, increased concentrations of one of these proteins negatively influenced the transcription stimulation mediated by the second regulator. This may suggest a competition between DnaA and SeqA for binding to the p(R) regulatory region. The physiological significance of this DnaA and SeqA-mediated regulation of p(R) is demonstrated by studies on lambda plasmid DNA replication in vivo. PMID- 12742019 TI - Structural basis for antibody catalysis of a cationic cyclization reaction. AB - Antibody 4C6 efficiently catalyzes a cationic cyclization reaction. Crystal structures of the antibody 4C6 Fab in complex with benzoic acid and in complex with its eliciting hapten were determined to 1.30A and 2.45A resolution, respectively. These crystal structures, together with computational analysis, have elucidated a possible mechanism for the monocyclization reaction. The hapten complex revealed a combining site pocket with high shape complementarity to the hapten. This active site cleft is dominated by aromatic residues that shield the highly reactive carbocation intermediates from solvent and stabilize the carbocation intermediates through cation-pi interactions. Modeling of an acyclic olefinic sulfonate ester substrate and the transition state (TS) structures shows that the chair-like transition state is favored, and trapping by water directly produces trans-2-(dimethylphenylsilyl)-cyclohexanol, whereas the less favored boat-like transition state leads to cyclohexene. The only significant change observed upon hapten binding is a side-chain rotation of Trp(L89), which reorients to form the base of the combining site. Intriguingly, a benzoic acid molecule was sequestered in the combining site of the unliganded antibody. The 4C6 active site was compared to that observed in a previously reported tandem cyclization antibody 19A4 hapten complex. These cationic cyclization antibodies exhibit convergent structural features with terpenoid cyclases that appear to be important for catalysis. PMID- 12742020 TI - Gelsolin domains 4-6 in active, actin-free conformation identifies sites of regulatory calcium ions. AB - Structural analysis of gelsolin domains 4-6 demonstrates that the two highest affinity calcium ions that activate the molecule are in domains 5 and 6, one in each. An additional calcium site in domain 4 depends on subsequent actin binding and is seen only in the complex. The uncomplexed structure is primed to bind actin. Since the disposition of the three domains is similar in different crystal environments, either free or in complex with actin, the conformation in calcium is intrinsic to active gelsolin itself. Thus the actin-free structure shows that the structure with an actin monomer is a good model for an actin filament cap. The last 13 residues of domain 6 have been proposed to be a calcium-activated latch that, in the inhibited form only, links two halves of gelsolin. Comparison with the active structure shows that loosening of the latch contributes but is not central to activation. Calcium binding in domain 6 invokes a cascade of swapped ion-pairs. A basic residue swaps acidic binding partners to stabilise a straightened form of a helix that is kinked in inhibited gelsolin. The other end of the helix is connected by a loop to an edge beta-strand. In active gelsolin, an acidic residue in this helix breaks with its loop partner to form a new intrahelical ion-pairing, resulting in the breakage of the continuous sheet between domains 4 and 6, which is central to the inhibited conformation. A structural alignment of domain sequences provides a rationale to understand why the two calcium sites found here have the highest affinity amongst the five different candidate sites found in other gelsolin structures. PMID- 12742021 TI - Elaborate manifold of short hydrogen bond arrays mediating binding of active site directed serine protease inhibitors. AB - An extensive structural manifold of short hydrogen bond-mediated, active site directed, serine protease inhibition motifs is revealed in a set of over 300 crystal structures involving a large suite of small molecule inhibitors (2-(2 phenol)-indoles and 2-(2-phenol)-benzimidazoles) determined over a wide range of pH (3.5-11.4). The active site hydrogen-bonding mode was found to vary markedly with pH, with the steric and electronic properties of the inhibitor, and with the type of protease (trypsin, thrombin or urokinase type plasminogen activator (uPA)). The pH dependence of the active site hydrogen-bonding motif is often intricate, constituting a distinct fingerprint of each complex. Isosteric replacements or minor substitutions within the inhibitor that modulate the pK(a) of the phenol hydroxyl involved in short hydrogen bonding, or that affect steric interactions distal to the active site, can significantly shift the pH-dependent structural profile characteristic of the parent scaffold, or produce active site binding motifs unique to the bound analog. Ionization equilibria at the active site associated with inhibitor binding are probed in a series of the protease inhibitor complexes through analysis of the pH dependence of the structure and environment of the active site-binding groups involved in short hydrogen bond arrays. Structures determined at high pH (>11), suggest that the pK(a) of His57 is dramatically elevated, to a value as high as approximately 11 in certain complexes. K(i) values involving uPA and trypsin determined as a function of pH for a set of inhibitors show pronounced parabolic pH dependence, the pH for optimal inhibition governed by the pK(a) of the inhibitor phenol involved in short hydrogen bonds. Comparison of structures of trypsin, thrombin and uPA, each bound by the same inhibitor, highlights important structural variations in the S1 and active sites accessible for engineering notable selectivity into remarkably small molecules with low nanomolar K(i) values. PMID- 12742022 TI - The allosteric transition of GroEL induced by metal fluoride-ADP complexes. AB - To understand the mechanism of a functionally important ATP-induced allosteric transition of GroEL, we have studied the effect of a series of metal fluoride-ADP complexes and vanadate-ADP on GroEL by kinetic fluorescence measurement of pyrene labeled GroEL and by small-angle X-ray scattering measurement of wild-type GroEL. The metal fluorides and vanadate, complexed with ADP, are known to mimic the gamma-phosphate group of ATP, but they differ in geometry and size; it is expected that these compounds will be useful for investigating the strikingly high specificity of GroEL for ATP that enables the induction of the allosteric transition. The kinetic fluorescence measurement revealed that aluminium, beryllium, and gallium ions, when complexed with the fluoride ion and ADP, induced a biphasic fluorescence change of pyrenyl GroEL, while scandium and vanadate ions did not induce any kinetically observed change in fluorescence. The burst phase and the first phase of the fluorescence kinetics were reversible, while the second phase and subsequent changes were irreversible. The dependence of the burst-phase and the first-phase fluorescence changes on the ADP concentration indicated that the burst phase represents non-cooperative nucleotide binding to GroEL, and that the first phase represents the allosteric transition of GroEL. Both the amplitude and the rate constant of the first phase of the fluorescence kinetics were well understood in terms of a kinetic allosteric model, which is a combination of transition state theory and the Monod Wyman-Changeux allosteric model. From the kinetic allosteric model analysis, the relative free energy of the transition state in the metal fluoride-ADP-induced allosteric transition of GroEL was found to be larger than the corresponding free energy of the ATP-induced allosteric transition by more than 5.5kcal/mol. However, the X-ray scattering measurements indicated that the allosteric state induced by these metal fluoride-ADP complexes is structurally equivalent to the allosteric state induced by ATP. These results suggested that both the size and coordination geometry of gamma-phosphate (and its analogs) are related to the allosteric transition of GroEL. It was therefore concluded that the tetrahedral geometry of gamma-phosphate (or its analogs) and the inter-atomic distance ( approximately 1.6A) between phosphorus (vanadium, or metal atom) and oxygen (or fluorine) are both important for inducing the allosteric transition of GroEL, leading to the high selectivity of GroEL for ATP about ligand adenine nucleotides, which function as the preferred allosteric ligand. PMID- 12742023 TI - NMR solution structure of the non-RGD disintegrin obtustatin. AB - The solution structure of obtustatin, a novel non-RGD disintegrin of 41 residues isolated from Vipera lebetina obtusa venom, and a potent and selective inhibitor of the adhesion of integrin alpha(1)beta(1) to collagen IV, has been determined by two-dimensional nuclear magnetic resonance. Almost the whole set of chemical shifts for 1H, 13C and 15N were assigned at natural abundance from 2D homonuclear and heteronuclear 500 MHz, 600 MHz and 800 MHz spectra at pH 3.0 recorded at 298 K and 303 K. Final structural constraints consisted of 302 non-redundant NOE (95 long-range, 60 medium, 91 sequential and 56 intra-residue), four disulfide bond distances, five chi1 dihedral angles and four hydrogen bonds. The 20 conformers with lowest total energy had no NOE violations greater than 0.35A or dihedral angle violations greater than 12 degrees. The average root-mean-square deviation (RMSD) for backbone atoms of all residues among the 20 conformers was 1.1A and 0.6A for the 29 best-defined residues. Obtustatin lacks any secondary structure. Compared to all known disintegrin structures in which the RGD motif is located at the apex of an 11 residue hairpin loop, the active KTS tripeptide of obtustatin is oriented towards a side of its nine residue integrin-binding loop. The C terminal tail is near to the active loop, and these two structural elements display the largest atomic displacements due to local conformational disorder. Double cross-peaks for W20, Y28 and H27 in the aromatic region of TOCSY spectra, local RMSD values for these residues, and positive cross-peaks in a ROESY spectrum (600 MHz, 100 ms mixing time), suggest that these residues act as a hinge allowing for the overall flexibility of the entire integrin-binding loop. These distinct structural features, along with its different electrostatic surface potential in relation to other known disintegrins, may confer to obtustatin its reported alpha(1)beta(1) integrin inhibitory selectivity. PMID- 12742024 TI - Crystal structure of the avilamycin resistance-conferring methyltransferase AviRa from Streptomyces viridochromogenes. AB - The emergence of antibiotic-resistant bacterial strains is a widespread problem in contemporary medical practice and drug design. It is therefore important to elucidate the underlying mechanism in each case. The methyltransferase AviRa from Streptomyces viridochromogenes mediates resistance to the antibiotic avilamycin, which is closely related to evernimicin, an oligosaccharide antibiotic that has been used in medical studies. The structure of AviRa was determined by X-ray diffraction at 1.5A resolution. Phases were obtained from one selenomethionine residue introduced by site-directed mutagenesis. The chain-fold is similar to that of most methyltransferases, although AviRa contains two additional helices as a specific feature. A putative-binding site for the cofactor S-adenosyl-L methionine was derived from homologous structures. It agrees with the conserved pattern of interacting amino acid residues. AviRa methylates a specific guanine base within the peptidyltransferase loop of the 23S ribosomal RNA. Guided by the target, the enzyme was docked to the cognate ribosomal surface, where it fit well into a deep cleft without contacting any ribosomal protein. The two additional alpha-helices of AviRa filled a depression in the surface. Since the transferred methyl group of the cofactor is in a pocket beneath the enzyme surface, the targeted guanine base has to flip out for methylation. PMID- 12742026 TI - Asymmetry in the triple helix of collagen-like heterotrimers confirms that external bonds stabilize collagen structure. AB - Heating and subsequent cooling mixtures of (Pro-Pro-Gly)(10) and (Pro-Hyp Gly)(10) peptides leads to formation of model heterotrimeric collagen helices that can be isolated by HPLC. These heterotrimeric collagen peptide helices are shown to be fundamentally unstable as denaturing then renaturing experiments result in heterotrimeric/homotrimeric mixtures. As the proportion of hydroxyproline-containing chains in the trimers increases, differential scanning calorimetry shows that the helix melting temperatures and denaturation enthalpies increasing non-linearly. Three types of Rich-Crick hydrogen bonds observed by NMR allow modelling of heterotrimeric structures based on published homotrimeric X ray data. This revealed a small axial movement of (Pro-Hyp-Gly)(10) chains towards the C-terminal of the helix, demonstrating heterotrimeric asymmetry. PMID- 12742028 TI - Mixed adsorption of poly(vinylpyrrolidone) and sodium dodecylbenzenesulfonate on kaolinite. AB - The mixed adsorption of the nonionic polymer poly(vinylpyrrolidone) (PVP) and the anionic surfactant sodium dodecylbenzenesulfonate (SDBS) on kaolinite has been studied. Both components adsorb from their mixture onto the clay mineral. The overall adsorption process is sensitive to the pH, the electrolyte concentration, and the amounts of polymer and surfactant. Interpretation of the experimental data addresses also the patchwise heterogeneous nature of the clay surface. In the absence of PVP, SDBS adsorbs on kaolinite by electrostatic and hydrophobic interactions. However, when PVP is present, surfactant adsorption at 10(-2) M NaCl is mainly driven by charge compensation of the edges. The adsorption of PVP from the mixture shows similar behavior under different conditions. Three regions can be distinguished based on the changing charge of polymer-surfactant complexes in solutions with increasing SDBS concentration. At low surfactant content, PVP adsorbs by hydrogen bonding and hydrophobic interactions, whereas electrostatic interactions dominate at higher surfactant concentrations. Over the entire surfactant concentration range, polymer-surfactant aggregates are present at the edges. The composition of these surface complexes differs from that in solution and is controlled by the surface charge. PMID- 12742025 TI - A novel approach to decoy set generation: designing a physical energy function having local minima with native structure characteristics. AB - We suggest a new approach to the generation of candidate structures (decoys) for ab initio prediction of protein structures. Our method is based on random sampling of conformation space and subsequent local energy minimization. At the core of this approach lies the design of a novel type of energy function. This energy function has local minima with native structure characteristics and wide basins of attraction. The current work presents our motivation for deriving such an energy function and also tests the derived energy function. Our approach is novel in that it takes advantage of the inherently rough energy landscape of proteins, which is generally considered a major obstacle for protein structure prediction. When local minima have wide basins of attraction, the protein's conformation space can be greatly reduced by the convergence of large regions of the space into single points, namely the local minima corresponding to these funnels. We have implemented this concept by an iterative process. The potential is first used to generate decoy sets and then we study these sets of decoys to guide further development of the potential. A key feature of our potential is the use of cooperative multi-body interactions that mimic the role of the entropic and solvent contributions to the free energy. The validity and value of our approach is demonstrated by applying it to 14 diverse, small proteins. We show that, for these proteins, the size of conformation space is considerably reduced by the new energy function. In fact, the reduction is so substantial as to allow efficient conformational sampling. As a result we are able to find a significant number of near-native conformations in random searches performed with limited computational resources. PMID- 12742029 TI - The structure of water near platinum and its significance in water-adsorbent system: molecular dynamics study. AB - Water structure around hydrophilic adsorbents is expected to differ from normal water structure. In some cases, a change of water structure seems to significantly affect phenomena involved in water-adsorbent systems. To investigate this, we have performed molecular dynamic simulations of three systems, water between (111) surfaces of Pt, water between (100) surfaces of Pt, and pure water at 298 K. For the analysis of water structure, we concentrated on the five-membered ring (R5) structure and six-membered ring (R6) structure, which are the most probable in nature. For both (111) and (100) of Pt, the ratio of hexameric ring/pentameric ring (R6/R5) in semibound regions was higher than that of bulk water, as expected. The function of adsorbent ceramics was explained in terms of the change of water structure when they are used as biomaterials, support materials, etc. The obscure mechanisms for the effects of medicine stones, antioxidants, etc., were also discussed from this point of view. PMID- 12742030 TI - Elucidation of adsorption mechanism of bone-staining agent alizarin red S on hydroxyapatite by FT-IR microspectroscopy. AB - To elucidate adsorption mechanism of alizarin red S (ARS), which is often used for staining bones in histology, adsorption of ARS on hydroxyapatite, Ca10(PO4)6(OH)2 (HAP), was investigated by a batch method, compared with alizarin, phenols, and benzenesulfonates. We found that ionized 1-, 2-OH groups (1-, 2-O(-)) of ARS can be electrostatically bound to Ca2+ on HAP, but that the 3 SO3(-) group of ARS hardly participates in adsorption on HAP. ARS-adsorbed HAP (ARS-HAP) in dark reddish violet was also prepared and analyzed by FT-IR microspectroscopy to gain structural information on bonding between ARS and HAP. The obtained spectrum, which was converted to difference spectra, indicated a single band of nu(C=O) at 1627 cm(-1) and two types of symmetric C=O stretching bands of nu(s)(C=O) + nu(C=C) at 1345 cm(-1) and nu(s)(C=O) + delta(O-C=C) at 1272 cm(-1). These bands imply the existence of a salt form in ARS-HAP via 1-, 2 OH groups of ARS. As a result of the existence of a chelate form in ARS-HAP via 1 OH and 9-C=O groups of ARS, two bands of nu(C=C) + nu(C=O) at 1572 cm(-1) and nu(C=O) + nu(C=C) at 1537 cm(-1) were also observed. In addition, ARS was almost desorbed from colored ARS-HAP at 50 degrees C by using neutral phosphate buffer to recover slightly pale pinkish HAP, or De-ARS-HAP. The desorbed ARS belongs to ARS previously adsorbed on HAP by salt formation, while the remaining color on De ARS-HAP indicates ARS still adsorbed on HAP by chelate formation. Consequently, we elucidated two adsorption mechanisms of ARS on HAP: The major adsorption is salt formation made up with 1-, 2-O(-) of ARS and Ca2+ on HAP, and the minor adsorption is chelate formation made up with 1-O(-) and 9-C=O of ARS and Ca2+ on HAP. PMID- 12742031 TI - Water-vapor adsorption and surface area measurement of poorly crystalline boehmite. AB - Water-vapor adsorption on poorly crystalline boehmite (PCB) was studied using a gravimetric FTIR apparatus that measured FTIR spectra and water adsorption isotherms simultaneously. The intensity of the delta(HOH) band of adsorbed water changed linearly with water content and this linear relationship was used to determine the dry mass of the sample. Adsorption and desorption isotherms of PCB showed a Type IV isotherm. The BET(H2O) surface area of PCB was 514+/-36 m2/g. The mean crystallite dimensions of PCB were estimated to be 4.5 x 2.2 x 10.0 nm (dimensions along the a, b, and c axes, respectively) based on application of the Scherrer equation to powder diffraction data of PCB. A surface area value of 504+/-45 m2/g calculated using the mean crystallite dimensions was in good agreement with the BET(H2O) surface area. This work also demonstrated a method to determine surface areas for materials with minimal perturbation of their surface structure. In addition, the FTIR spectra of PCB were influenced by changes in water content. The delta(AlOH) band at 835 cm(-1) observed under dry conditions was assigned to the non-H-bonded surface OH groups. As the amount of adsorbed water increased, the intensity at 835 cm(-1) decreased and that at 890 and 965 cm(-1) increased. The 890- and 965-cm(-1) bands are assigned to surface OH groups H-bonded with adsorbed water. PMID- 12742032 TI - Multiscale simulation of irreversible deposition in presence of double layer interactions. AB - Sequential lattice Monte Carlo simulations, in which the transition probabilities are derived from the discrete form of the continuum-level mass conservation law, are used to predict the morphology of colloidal deposits. The simulations account for particle-surface (P-S) and particle-particle (P-P) electrostatic and van der Waals interactions. Simulation results for maximum coverage for monolayer deposition are in quantitative agreement with the hard-sphere RSA jamming limit. Moreover, as reported in earlier studies, monolayer simulations in the absence of P-S interactions qualitatively predict the monotonic increases in fractional coverage with increasing ionic strength, characterized by the Debye screening length (kappa a). Monolayer simulations with P-S interactions show that the dependence of fractional coverage on kappa a is strongly influenced by the ratio of particle to surface potentials (Psi(p)/Psi(s)). P-S and P-P forces achieve their respective maximum at different values of kappa a leading to a nonmonotonic trend in surface coverage as a function of kappa a. These results indicate that the incorporation of P-S interactions into colloidal deposition studies allows more accurate interpretation of the experimental data. In multilayer deposition simulations, balance between long-ranged weak interactions and short-ranged strong interactions between P-P and P-S, coupled with physical screening effects, resulted in widely varying coverages with height of the deposit, ionic strength, and Psi(p)/Psi(s). Moreover, fractal dimension of the deposit ranged from approximately 1 (kappa a << 1) to 1.7 (kappa a >> 1). Qualitative kinetic analysis showed widely varying deposition rates in different layers depending on Psi(p)/Psi(s) and ionic strength. The multilayer system approached the monolayer system in the limit kappa a--> infinity and Psi(p)/Psi(s)--> infinity. PMID- 12742033 TI - Molecular interactions between DNA and an aminated glass substrate. AB - With the development of DNA arrays, the immobilization of DNA strands onto solid substrates remains an essential research topic. DNA arrays have potential applications in DNA sequencing, mutation detection, and pathogen identification. DNA bound to solid substrates must still be accessible and retain the ability to hybridize with its complementary strands. One technology to produce these arrays involves linking DNA molecule probes to a silanized substrate in microspot patterns and exposing them to a solution of fluorescently labeled samples of DNA targets. The behavior of both the target and probe DNA and their interactions with each other at the substrate surface, particularly with respect to molecular interactions, are poorly understood at the present time. The objective of this work is to model simply the interface interactions between DNA and glass slides modified with an aminosilane (gamma-aminopropyltriethoxysilane, APTS). In aqueous solutions, DNA behaves as a polyacid over a wide range of pH. A glass substrate treated with APTS is positively or negatively charged, depending on the pH. A model of the surface charge of APTS-treated glass has been developed from results of wetting experiments performed at various pH. It has been demonstrated that the surface charge of APTS-treated glass is well described by a model of constant capacitance of the electrical double layer. A good correlation between experimental data on DNA retention at various pH's and the variation of the surface charge of the APTS-treated glass is obtained. This provides an indication of the role of ionic interactions in the adsorption of DNA molecules onto aminated glass slides. PMID- 12742034 TI - Study of interaction of proteins with fumed silica in aqueous suspensions by adsorption and photon correlation spectroscopy methods. AB - The interaction of fumed silica A-300 (S(BET) = 297 m2 g(-1)) with bovine serum albumin (prepared by different methods), ovalbumin, human hemoglobin, and gelatin as a function of pH, salinity, and concentrations of components in aqueous medium was studied by adsorption and photon correlation spectroscopy (PCS) methods. Comparison of equilibrium (incubation time t(i) approximately 1 h) adsorption of proteins on A-300, minute (t(i) approximately 1 min) flocculation rate, and the particle size distributions measured by the PCS method shows different rearrangement of particle swarms depending on pH, salinity, and concentration of proteins, especially at pH close to IEP of silica or proteins. The electrokinetic mobility of protein/silica swarms is greater than that of individual components at pH far from the IEP of proteins. Changes in the Gibbs free energy (DeltaG) on protein adsorption depend on pH (-DeltaG is minimal at pH 2, close to the IEP of silica, and maximal at pH between the IEP of protein and silica), concentration ( DeltaG is maximal at C(p) between 1 and 6 mg/ml), type of proteins, and their preparation technique. PMID- 12742035 TI - Oxidation behavior of Ru/TiO2 and metallic Ru fine particles on heating in air. AB - Thermogravimetry (TG) and differential thermal analysis (DTA) were used to investigate the oxidation behavior of Ru/TiO2 and metallic Ru fine particles during heating in air in the range 20-1000 degrees C. Temperature ranges of the oxidation for two samples of Ru/TiO2 with the compositions (92 wt% Ru, 8 wt% TiO2) and (75 wt% Ru, 25 wt% TiO2) and for pure metallic Ru fine particle agglomerates were determined. It was assumed that after the partial oxidation of Ru in the sample containing 75 wt% Ru and 25 wt% TiO2 and in the pure metallic Ru a diffusion barrier was formed, preventing further oxidation of Ru in Ru/RuO2 and Ru/RuO2/TiO2 matrices. XRD and TEM were used for the sample characterization. PMID- 12742036 TI - Preparation of highly concentrated stable dispersions of uniform silver nanoparticles. AB - Stable concentrated aqueous dispersions of silver nanoparticles of narrow size distribution were prepared by reducing silver nitrate solutions with ascorbic acid in the presence of Daxad 19 (sodium salt of a high-molecular-weight naphthalene sulfonate formaldehyde condensate) as stabilizing agent. The latter has excellent ability to prevent the aggregation of nanosize silver at high ionic strength and high concentration of metal (up to 0.3 mol dm(-3)). The presence of the dispersing agent on the surface of silver particles was confirmed by ATR-FTIR spectroscopy and electrokinetic measurements, explaining both the negative charge over the entire pH range and the electrosteric effect responsible for their long term stability. The other experimental conditions, i.e., the pH of the reacting solutions, the concentration of the stabilizing agent, and the metal/dispersant ratio, also have a significant impact on the size and stability of these dispersions. The final nanosize silver can be obtained as dried powder, and can be fully redispersed in deionized water by sonication. PMID- 12742037 TI - In situ preparation of weakly flocculated aqueous anatase suspensions by a hydrothermal technique. AB - Weakly flocculated aqueous anatase suspensions were prepared in situ by hydrothermally treating amorphous titania particles peptized with different amounts of tetraethylammonium hydroxide (TENOH). The simultaneous formation of hydrous TiO2 polyanions in the presence of OH- and tetraethylammonium cations are two essential conditions for the peptization process to occur. The absence of either of these conditions will cause reprecipitation. Transmission electron microscopy (TEM) revealed that the morphology of the particles formed at low TENOH concentrations consisted of well-dispersed anatase crystals, changing to asterisk-like structured particles with increasing concentrations of TENOH. Because of the extremely high absolute zeta potential (over -70 mV in all the samples) and ionic strength values, nontouching particle networks may be formed in situ in the mother solution in all samples, as predicted by DLVO theory. A trend to coagulation was observed in the suspensions with increasing concentrations of TENOH due to a more pronounced secondary minimum in the particle pair potential curves. Assuming the particles remained in the secondary minimum throughout the hydrothermal treatment may lead to the formation of the asterisk-like hard agglomerates. This may arise from the condensation of the -OH rich TiO2 particles or from the deposition of material in the interparticle gap during the particle growth process. The green packing density of slip-cast bodies from a suspension containing 20 wt% solids was around 46%. PMID- 12742038 TI - Production of aqueous colloidal dispersions of carbon nanotubes. AB - Stable homogeneous dispersions of carbon nanotubes (CNTs) have been prepared by using sodium dodecyl sulfate (SDS) as dispersing agent. To our knowledge, it is the first report to quantitatively characterize colloidal stability of the dispersions by UV-vis spectrophometric measurements. When the sediment time reaches 500 h, the supernatant CNT concentration drops as much as 50% for the bare CNT suspension, compared to 15% with the addition of SDS. Furthermore, after 150 h, no precipitation is found for CNT/SDS dispersions, exhibiting an extreme stability. Zeta potential, auger electron microscopy, and FTIR analysis are employed to investigate the adsorption mechanism in detail. It has been concluded that the surfactant containing a single straight-chain hydrophobic segment and a terminal hydrophilic segment can modify the CNTs-suspending medium interface and prevent aggregation over long periods. The morphology of the CNT dispersions is observed with optical microscopy. An intermediate domain of homogeneously dispersed nanotubes exhibits an optimum at 0.5 wt% CNTs and 2.0 wt% SDS. PMID- 12742039 TI - Faradaic depolarization in the electrokinetics of the metal-electrolyte solution interface. AB - Streaming potentials (E(str)) have been measured in a flat thin-layer cell with gold and aluminum surfaces. The conventional relation between E(str) and the zeta potential is shown to be applicable only as long as charge transfer reactions at the metal-electrolyte solution interface are insignificant in terms of the ensuing contribution to the overall cell conductivity. Owing to the irreversibility of the reduction/oxidation of water at most metal surfaces, streaming potentials can be obtained over a very broad range of pressure gradients for metallic substrates in electrolytes such as KNO3. The situation changes drastically in the presence of a reversible redox couple like Fe(CN)(6)3 /Fe(CN)(6)4-. Even small streaming potentials are then greatly diminished due to the extensive conduction that results from the bipolar electrolysis at the metal surface. For gold and aluminum in the presence of various electroinactive and electroactive electrolytes, the measured values for E(str) are shown to be consistent with their conventional voltammetric characteristics. PMID- 12742040 TI - Numerical calculation of the electrophoretic mobility of colloidal particles in weak electrolyte solutions. AB - A numerical calculation of the electrophoretic mobility of colloidal particles in weak electrolyte solution is presented. It is based on a previous work (C. Grosse, V.N. Shilov, J. Colloid Interface Sci. 211 (1999) 160-170), where the analytical theory of the thin double layer concentration polarization is generalized to the case of weak electrolytes, i.e., when the dissociation recombination equilibrium and rate constants both have finite values. The analytical results are first completed by including terms corresponding to co ions that were neglected in the original presentation. It is shown that these terms that have little bearing in the case of strong electrolytes, become quite important in the case when the electrolyte is weak. The problem is then solved using the network method, leading to numerical results for the electric potential and the concentrations of counterions, co-ions, and neutral ion pairs. Finally, the electrophoretic mobility is calculated both analytically and numerically. It is shown that the hypothesis of a weak electrolyte leads to changes of mobility with respect to the classical results that are even stronger than predicted analytically. PMID- 12742041 TI - Dynamic electrophoretic mobility of a sphere in a spherical cavity. AB - Dynamic electrophoresis is a powerful analytical tool for the description of the surface properties of the charged entities in a concentrated dispersion. In our study the boundary effect on this dynamic phenomenon is investigated theoretically for the case, when the surface potential is low. In particular, the dynamic electrophoresis of a sphere in a spherical cavity is discussed as are the effects of the key factors on the phenomenon under consideration, which include the thickness of the double layer, the frequency of the applied electric field, the ratio of particle radius to cavity radius, and the boundary conditions of the surfaces of the particle and the cavity. The results of numerical simulation reveal that these key factors can have both quantitative and qualitative influence on the electrophoretic behavior of the particle. As an example, for the case of a positively charged particle placed in a negatively charged cavity if the double layer surrounding the particle is thin, the magnitude of the electrophoretic mobility of the particle increases with an increase in the frequency of the applied electric field and a phase lead may occur, but the opposite is true if the double layer is thick. These effects are not observed for the case of a positively charged particle placed in an uncharged cavity or for a positively charged particle placed in a positively charged cavity. PMID- 12742042 TI - Microscopic structure and properties of an interface between coexisting phases of an associating fluid adsorbed in slitlike pores. AB - Using density functional theory, we calculate density profiles of an associating fluid in slitlike pores. These profiles characterize an interface between two coexisting, adsorbed phases, e.g., between gaseous and liquid phases formed during capillary condensation. Our study has been carried out for weakly, as well as for strongly, associated fluids confined in pores of different widths. We also investigate the role of the fluid-wall interaction. PMID- 12742043 TI - Retardation of water evaporation by less-defective mixed monolayers spread from bulk solids onto water surface. AB - From AFM observation of transferred films on mica, it has been found that mixed monolayers of hexadecanol with poly(vinyl stearate) give a film with a less defective and flat surface by spreading from bulk solids of those mixtures onto a water surface without using any organic solvent. As a result, those mixed monolayers have a considerably larger effect on retardation of water evaporation in comparison with those spread from the solution of the mixtures. After the saturated surface pressure of the mixed monolayer spread from the bulk solids, an enhanced effect on retardation of water evaporation was found, accompanied by the preferential spreading of hexadecanol and the gradual reduction of defects in the mixed monolayer. PMID- 12742044 TI - Second harmonic generation from insoluble films of a rhodamine dye at the air water interface: effect of sodium dodecylsulfate. AB - The second harmonic generation (SHG) from the insoluble monolayers of bis-(N ethyl, N-octadecyl)rhodamine perchlorate (RhC18) formed on the surface of sodium dodecylsulfate (SDS) solutions of different concentrations has been studied. An enhancement of the second harmonic response was observed in the mixed films of RhC18/SDS compared to the pure-dye layer. To clarify the origin of the phenomenon, the films were characterized by surface pressure-area isotherm and reflection-absorption spectroscopy studies. The analysis of surface pressure-area isotherms of RhC18/SDS layers showed that incorporated SDS molecules essentially influence the rheological properties of the dye monolayer. The film parameters, such as the molecular surface area, maximum surface pressure, and solid-condensed phase composition, are the functions of SDS bulk concentration. A joint analysis of the SHG results and the reflection-absorption spectra revealed that the structural ordering within films was responsible for the enhancement of the nonlinear optical response, whereas the contributions from the spectral shifts and increased absorption upon aggregate formation are of less importance. PMID- 12742045 TI - Linear chains and chain-like fractals from electrostatic heteroaggregation. AB - The internal structure of materials prepared by aggregation of oppositely charged polystyrene spheres (electrostatic heteroaggregation) is investigated by static light scattering, optical microscopy, and Brownian dynamics simulation. Light scattering indicates ultralow mass fractal dimensions, as low as 1.2. Such low fractal dimensions, approaching the theoretical limit of a linear object, imply a chaining mechanism. Optical micrographs reveal linear chains with the particle charge alternating down the chains. Brownian dynamics simulation gives additional support for a chaining mechanism. For the polystyrene system (120-nm primary particle diameters), the fractal dimension is found to increase from 1.2 to 1.7 as the background electrolyte is increased. In terms of electrostatic screening, the results match those reported recently for larger polystyrene spheres. The low fractal dimensions appear to represent a crossover from linear chains to a structure of diffusion-limited aggregates; however, experiments under density neutral conditions imply that sedimentation plays an important role in the formation of ultralow fractal dimensions. The practical implication is that microcomposites with a locally uniform distribution of starting materials and almost any degree of branching can be prepared from oppositely charged particles. PMID- 12742046 TI - Morphology control of gamma-Fe2O3 nanocrystals via PEG polymer and accounts of its Mossbauer study. AB - Granular, needle-, rod-, or whisker-shaped gamma-Fe2O3 nanocrystals have been prepared via controlling the chemical microenvironment of the reaction system under gamma-irradiation. A novel extension-curl-effect model was proposed to explain the effect of the chemical microenvironment on the morphology of gamma Fe2O3 nanocrystals. The as-prepared gamma-Fe2O3 nanocrystals were characterized by powder X-ray diffraction and transmission electron microscopy. Their magnetic properties were also studied by zero-field Mossbauer spectroscopy, which indicated that different shaped gamma-Fe2O3 nanocrystals could exhibit interesting magnetic behavior. PMID- 12742047 TI - Interaction of the zeolitic tuff with Zn-containing simulated pollutant solutions. AB - The possibility of removing Zn2+ cations from wastewater by ion exchange using natural zeolites as exchangers has been investigated. The process of binding of zinc ions into zeolite structure has been established by several reaction mechanisms as a fast chemical reaction of ion exchange, accompanied by slower adsorption of different ionic species and possible precipitation or coprecipitation with the zeolite structure. The physicochemical phenomena such as hydrolysis and dissolution of surface layers are the result of interaction of zeolite with hydrogen or hydroxyl ions from the solution. Complexation of OH- with Zn2+ to form the zinc-hydroxy species strongly depends on pH value and affect the uptake mechanism as to lower dissolution of surface aluminosilicate layers. Structure imperfections as a surface property of mineralogical nonhomogeneous zeolitic grains can lead to formation of sorption surface sites with different energy, which affects the nonuniform distribution of different zinc species adsorbed. It is particularly possible in zeolitic tuff samples with relatively high content of aluminosilicates as minor mineralogical components, which is characteristic of Croatian deposits. PMID- 12742048 TI - Acidity distribution of carboxyl groups in Loy Yang brown coal: its analysis and the change by heat treatment. AB - Brown coals have a considerable number of acidic functional groups of which the main component is carboxyl groups, and the acidity has a wide distribution. In this paper, changes of the acidity distribution were examined by aqueous titration when brown coal was heat-treated to control its acidity distribution. For Loy Yang brown coal from Australia dried at 50 degrees C under vacuum (LY50), the acid dissociation constant, Ka, was distributed over a wide pKa range between 2 and 9. Then, using Gaussian functions, the acidity distribution was divided into four groups, which were characterized by average pKa values: average pKa value of 3.8 (hereafter referred to as Group A), 5.2 (Group B), 6.8 (Group C), and 8.3 (Group D). Among them, Groups A, B, and C were assigned to carboxyl groups. From the changes of the number of carboxyl groups when brown coal was heat-treated up to 400 degrees C, it was found that the way of decrease was different among these acidic groups. The decrease of the amount of carboxyl groups in Group C was significant, and at 325 degrees C most of them disappeared. On the other hand, the carboxyl groups in Group A remained even at a high temperature of 400 degrees C. We estimated approximately the structures around carboxyl groups for LY50 and their structural changes by heat treatment using the known pKa values for simple carboxylic acids and the pKa values calculated by the MOPAC program for complicated carboxylic acids. PMID- 12742049 TI - Sorption and diffusion of phenols onto well-defined ordered nanoporous monolithic silicas. AB - The sorption of phenol, and o-, m-, and p-aminophenol (o-, m-, and p-AP) onto highly ordered mesoporous silicas (HOM) with cubic Im3m (HOM-1), hexagonal H(I) (HOM-2), 3-D hexagonal p6(3)/mmc (HOM-3), cubic Ia3d (HOM-5), lamellar L( infinity ) (HOM-6), and solid phase S (HOM-8) materials has been investigated kinetically. Nanostructured silica molecular sieves have been prepared at 25 and 60 degrees C with lyotropic liquid-crystalline phases of the nonionic surfactant (Brij 76) that was used as a structure-directing agent. Such nanostructured silicas have been studied by 29Si nuclear magnetic resonance (29Si NMR), powder X ray diffraction (XRD), the Brunauer-Emmett-Teller (BET) method for nitrogen adsorption and surface area measurements, and transmission electron microscopy (TEM) techniques after synthesis and sorption. It was found that all materials exhibit well-defined long-range porous architectures without significant loss of the ordered texture during phenol sorption. The kinetics of phenol sorption has been studied spectrophotometrically at different temperatures (25-40 degrees C; +/-0.1 range). The sorption rate is zero order in all phenols sorbed, and increases directly in the pattern P >m-AP > o-AP > p-AP, which reflects the mobility of the phenol compounds on the particle pores. The isothermal sorption and the kinetic parameters were discussed and it was established that a diffusion controlled process characterizes phenol sorption. Furthermore, the mechanism of phenol sorption was deduced to be predominantly particle diffusion. The diffusion coefficients were determined using Fick's equation. The trend of diffusion of all phenols onto nanoporous silica was HOM-8 > HOM-2 > HOM-6 > HOM-5 > HOM-1 > HOM-3, reflecting the effect of the uniform pore size distribution and the internal surface area of the nanostructured silicas on the diffusion process. PMID- 12742050 TI - Transport properties of compact clays. II. Diffusion. AB - An experimental system was constructed in order to measure the diffusion coefficient in three types of porous media, namely mica, sodic montmorillonite, and natural compact clay. Several salts at various concentrations were used for the measurements in order to investigate the influence of these factors. Influence of porosity was also studied. In a first approximation, all the results can be summarized by a simple Archie's law independent of the clay and of the solute. The diffusion and electric formation factors have also been systematically compared; they generally agree for large porosities, while they disagree for small porosities for clay and montmorillonite. PMID- 12742051 TI - Polymer coatings to passivate calcite from acid attack: polyacrylic acid and polyacrylonitrile. AB - The extent of passivation of calcite toward dissolution by aqueous acids arising from polymeric coatings based on polyacrylic acid or polyacrylonitrile is evaluated using a channel flow cell technique with microdisc electrode detection. In situ passivation with polyacrylic acid leads to a reduction in the reactivity of calcite toward acid attack with a reduction in the rate constant by up to an order of magnitude compared with untreated calcite. Ex situ passivation with polyacrylic acid for 24 h results in good coverage of the calcite by the polymer but it is shown to erode from the surface when exposed to an aqueous acid solution. In contrast, polyacrylonitrile is demonstrated to form a regular coating after exposure for just 1 h and offers robust potent protection from aqueous acid attack. PMID- 12742052 TI - Investigation of the dependence of inferred interfacial tension on rotation rate in a spinning drop tensiometer. AB - The spinning drop tensiometer is widely used to study the interfacial properties of many systems. However, there have also been several reported limitations with the spinning drop tensiometer. In this paper, it is shown that there is a relationship between the measured interfacial tension and the rotation rate of the drop. A detailed investigation of this relationship is presented. PMID- 12742053 TI - A linear relation between the cloud point and the number of oxyethylene units of water-soluble nonionic surfactants valid for the entire range of ethoxylation. AB - The following linear equation correlates the cloud point (CP) of water-soluble polyoxyethylated nonionic surfactants (NSs) with the average number p of oxyethylene units per molecule: (p - p0)/CP = a + b(p - p0). Here p0 is the smallest value of p that confers solubility in cold water: In a homologous series of NSs, it belongs to the surfactant with CP = 0 degrees C. Plots of CP versus p for five representative homologous series of NSs consist of three segments: A steeply ascending, nearly straight line, a transition region that ranges from p = 15-22 to p = 20-28, and a nearly horizontal plateau that approaches asymptotically the CPs of polyethylene glycols with molecular weights between 30,000 and 4400. These CPs range from 113 to 130 degrees C. Most CPs for NSs were taken from the literature or measured on commercially available samples; eight CPs above 100 degrees C were measured on newly synthesized surfactants. Previously published linear equations correlating CP with p cover only NSs with p < 16 and CPs < 100 degrees C: They apply only to the ascending segment of the CP versus p plots. Our equation covers the entire plots and applies to the full range of NSs, including extensively polyoxyethylated NSs with p > or = 100. It can be used for selecting specific NSs for high-temperature applications. The hydrophile-lipophile balance of the surfactant with p = p0 oxyethylene units, namely, HLB0, is a novel quantitative measure of the hydrophobicity of the hydrocarbon moiety of the relevant homologous NS series. Its value reflects the size, composition, and structure of the hydrocarbon moiety. PMID- 12742054 TI - Effect of liquid slip in electrokinetic parallel-plate microchannel flow. AB - Liquid slip at hydrophobic surfaces in microchannels has frequently been observed. We present here an analytical solution for oscillating flow in parallel plate microchannels by combining the electrokinetic transport phenomena with Navier's slip condition. Our parametric results suggest that electrokinetic transport phenomena and liquid slip at channel walls are both important and should be considered simultaneously. Their significance depends on channel wall material, electrolyte concentration, and pH. For pressure-driven-flow, liquid slip counteracts the effect by the electrical double layer and induces a larger flow rate. A higher apparent viscosity would be predicted if slip is neglected. For electroosmotic flow, liquid slip alters the flow rate by about 20% for a thick electrical double layer. Our results provide design guidelines to precisely control time-dependent microflow in hydrophobic microfluidic microelectromechanical system devices. PMID- 12742055 TI - Surface energy of ethylene-co-1-butene copolymers determined by contact angle methods. AB - Wilhelmy plate measurements of contact angles with a series of test liquids are used to calculate the surface energies of two poly(ethylene-co-1-butene) random copolymers. Results from five methods of calculation are reported: one-liquid (Good-Girifalco and Neumann), two-liquid (harmonic mean and geometric mean), and three-liquid (Lifshitz-van der Waals acid-base) methods. We find that all five methods are sensitive to the choice of test liquids used for contact angle measurements, as previously reported, but consistent results are obtained if recommended combinations of liquids are used. The mean results of the three liquid acid-base method are judged to be the most reliable and informative, leading to surface energies of 30.8 mJ/m2 for poly(ethylene-co-1-butene) copolymer composed of 92 mol% ethylene and 30.2 mJ/m2 for copolymer composed of 88 mol% ethylene. PMID- 12742056 TI - Direct sonochemical preparation of high-surface-area nanoporous ceria and ceria zirconia solid solutions. AB - For the first time, nanoporous ceria and ceria-zirconia solid solutions with high surface area have been successfully synthesized directly via high-intensity ultrasound irradiation without thermal post-treatment. The ceria and the solid solutions were characterized by XRD, TEM, and nitrogen adsorption. It was found that the nanoporous structures of the materials obtained were formed by the agglomeration of monodispersed nanoparticles under high-intensity ultrasound irradiation. PMID- 12742057 TI - A simplified method for predicting the dynamic surface tension of concentrated surfactant solutions. AB - A simplified method for predicting the dynamic surface tension of concentrated surfactant solutions is proposed. It is implemented using the framework of the Henry's Law analytical solution to the Ward and Tordai equation for diffusion controlled adsorption, with the necessary parameters being deduced from the measured equilibrium surface tension equation and a value for the surfactant monomer diffusivity. The method is tested by calculating the dynamic surface tension relaxations of aqueous C10E6 and C10E8 solutions over concentration ranges from well below to well above their critical micelle concentrations (cmc). Results are compared with measured relaxations over 0.001-50 s, and semiquantitative agreement is found, with the best results obtained for concentrations near the cmc. The predictive method may prove useful in such applications as the screening of candidate surfactants for inks used in inkjet printing. PMID- 12742059 TI - A complete inventory of fungal kinesins in representative filamentous ascomycetes. AB - Complete inventories of kinesins from three pathogenic filamentous ascomycetes, Botryotinia fuckeliana, Cochliobolus heterostrophus, and Gibberella moniliformis, are described. These protein sequences were compared with those of the filamentous saprophyte, Neurospora crassa and the two yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe. Data mining and phylogenetic analysis of the motor domain yielded a constant set of 10 kinesins in the filamentous fungal species, compared with a smaller set in S. cerevisiae and S. pombe. The filamentous fungal kinesins fell into nine subfamilies when compared with well characterized kinesins from other eukaryotes. A few putative kinesins (one in B. fuckeliana and two in C. heterostrophus) could not be defined as functional, due to unorthodox organization and lack of experimental data. The broad representation of filamentous fungal kinesins across most of the known subfamilies and the ease of gene manipulation make fungi ideal models for functional and evolutionary investigation of these proteins. PMID- 12742060 TI - A genetic map of the lettuce downy mildew pathogen, Bremia lactucae, constructed from molecular markers and avirulence genes. AB - The genetic map of Bremia lactucae was expanded utilizing 97 F(1) progeny derived from a cross between Finnish and Californian isolates (SF5xC82P24). Genetic maps were constructed for each parent utilizing 7 avirulence genes, 83 RFLP markers, and 347 AFLP markers, and a consensus map was constructed from the complete data set. The framework map for SF5 contained 24 linkage groups distributed over 835cM; the map for C82P24 contained 21 linkage groups distributed over 606cM. The consensus map contained 12 linkage groups with markers from both parents and 24 parent-specific groups. Six avirulence genes mapped to different linkage groups; four were located at the ends of linkage groups. The closest linkages between molecular markers and avirulence genes were 3cM to Avr4 and 1cM to Avr7. Mating type seemed to be determined by a single locus, where the heterozygote determined the B(2) type and the homozygous recessive genotype determined the B(1) type. PMID- 12742061 TI - Analysis of loss of pathogenicity mutants reveals that repeat-induced point mutations can occur in the Dothideomycete Leptosphaeria maculans. AB - Restriction enzyme mediated insertional mutagenesis using a plasmid, pUCATPH, that confers hygromycin resistance, generated loss-of-pathogenicity mutants of Leptosphaeria maculans, the fungus that causes blackleg disease of Brassica napus. Of 516 L. maculans transformants analysed, 12 were pathogenicity mutants. When eight of these mutants were crossed to an isolate that attacks B. napus, cosegregation of pUCATPH sequences and loss of pathogenicity was not observed, suggesting that these mutations were not linked to plasmid sequences. In seven of eight crosses analysed, progeny with the hygromycin resistance gene were hygromycin-sensitive. Sequence analysis of an amplified fragment of pUCATPH in six clones derived from one 'silenced' progeny showed mutation of GC to AT on one DNA strand, reminiscent of repeat-induced point mutation (RIP) in Neurospora crassa. One loss-of-pathogenicity mutant had pUCATPH inserted in the promoter of a gene with an open reading frame of 529 amino acids that had no database match. Reintroduction of a wild-type copy of the gene to this mutant restored the ability to form lesions on cotyledons of B. napus. PMID- 12742062 TI - A system for studying genetic changes in Candida albicans during infection. AB - Candida albicans is a diploid yeast with a dimorphic life history. It exists commensally in many healthy humans but becomes a potent pathogen in immunocompromised hosts. The underlying genetic mechanisms by which C. albicans switches from a commensal to a pathogenic form in the host are not well understood. To study the evolution of virulence in mammalian hosts, we used GAL1 as selectable marker system that allows for both positive and negative selection in selective media. We show that the deletion of one or both copies of GAL1 in the C. albicans genome does not change virulence in a systemic mouse model. We obtained estimates for the frequency of mitotic recombination at the GAL1 locus during systemic infection. Our observations suggest that genetic changes such as mitotic recombination and gene conversion occur at a high enough frequency to be important in the transition of C. albicans from a commensal to a pathogenic organism. PMID- 12742064 TI - Chromosomal polymorphism, syntenic relationships, and ploidy in the pathogenic fungus Paracoccidioides brasiliensis. AB - Pulsed field gel electrophoresis (PFGE) and DNA hybridization were used to establish and compare the electrophoretic karyotypes of 12 clinical and environmental Paracoccidioides brasiliensis isolates from different geographic areas. Gene mapping allowed the identification of synteny groups and the use of isolated whole chromosomal bands to probe chromoblots indicated the existence of repetitive sequences, contributing to a better understanding of the structure and organization of the fungus genome. This represents the first comparative mapping study among different isolates. The results are indicative of the existence of genetic differences among natural isolates. DNA content of DAPI-stained nuclei of each isolate was estimated by confocal microscopy. Comparison of the genome sizes estimated by PFGE with those calculated by microfluorometry indicated the possible existence of haploid and diploid (or aneuploid) isolates of the fungus. PMID- 12742065 TI - Depletion of polyubiquitin encoded by the UBI4 gene confers pleiotropic phenotype to Candida albicans cells. AB - We have studied the roles of polyubiquitin in Candida albicans physiology. Heterologous expression of the C. albicans polyubiquitin (UBI4) gene in a ubi4 Saccharomyces cerevisiae strain suppressed the mutant phenotype (hypersensitivity to heat shock). A heterozygous strain UBI4/Deltaubi4::hisG, obtained following the ura-blaster procedure, was used to construct a conditional mutant using a pCaDis derivative plasmid. By serendipity we isolated the UBI4 conditional mutant as well as a UBI4 mutant containing a non-functional MET3 promoter. Depletion of polyubiquitin conferred pleiotropic effects to mutant cells: (i) a limited increased sensitivity to mild heat shock; (ii) increased formation of colony morphology variants; and (iii) induction of hyphal and pseudohypal development. These results indicate that polyubiquitin in C. albicans is involved in the negative control of switching, as well as in maintaining the yeast cell morphology, probably by silencing mechanisms triggering the hyphal and pseudohyphal development in the absence of environmental inducers. PMID- 12742066 TI - White-cap mutants and meiotic apoptosis in the basidiomycete Coprinus cinereus. AB - Among many white-cap mutants of Coprinus cinereus, four distinct classes have been identified cytologically. Mutants of one class progress through meiosis normally but fail to sporulate; the defect is post-meiotic and it triggers apoptosis in the tetrad stage. Mutants of the other three classes have defects in meiotic prophase and these are: (1) those that assemble synaptonemal complexes (SCs) normally; (2) those that assemble axial elements (AEs) but not SCs; and (3) those that assemble neither AEs nor SCs even though the chromosomes are condensed and also paired. All three meiotic mutant classes arrest at meiotic metaphase I and the arrest triggers meiosis-specific apoptosis showing characteristic chromatin condensation, DNA fragmentation as shown by the TUNEL assay, cytoplasmic shrinkage, and finally total DNA degradation. Apoptosis is very cell type specific; it occurs only in the basidia while the neighboring somatic cells are perfectly healthy and the mushroom continues to develop and mature with very few basidiospores produced. The meiotic apoptosis in C. cinereus is under strict cell cycle control rather than at any time after defect; apoptosis is triggered only after entry to meiotic metaphase. It is intriguing to note that C. cinereus has two checkpoints for arrest and entry to apoptosis: one is meiotic at the metaphase I spindle checkpoint regardless of the time of defects, and one is post meiotic at the tetrad stage. This is in striking contrast to multiple checkpoint arrests and entries to meiotic apoptosis found in the mouse. PMID- 12742067 TI - Evidence that the appressorial development in barley powdery mildew is controlled by MAP kinase activity in conjunction with the cAMP pathway. AB - Development of the barley powdery mildew fungus involves the sequential formation of a primary germ tube, an appressorial germ tube, and an appressorium. Previously, we have shown that the cAMP pathway controls the emergence of the two germ tubes. Following identification of two MAP kinase genes in an EST database from developing conidia we studied the role of the MAP kinase pathway and its interaction with the cAMP pathway. Fungal MAP kinase activity increased rapidly during mildew development, reaching a maximum between 2 and 8h after inoculation. Sphingosine or PAF-16, activators of the MAP kinase pathway, increased activity and appressorial development whilst an inhibitor, PD 98059, decreased both. Studies on the interaction between the cAMP and MAPK pathways revealed that several effectors of the MAPK pathway had no effect on cAMP levels. However upstream effectors of the cAMP pathway, such as cholera toxin and pertussis toxin (activators of G(alpha) proteins) increased MAPK activities whereas downstream effectors such as forskolin (adenylyl cyclase activator) or H89 (PKA inhibitor) had no effect. Combined application of forskolin and sphingosine produced a rise in appressorial germ tube and appressorial formation higher than when either pathway was stimulated individually. These results suggest that the two pathways cooperate in appressorial development. PMID- 12742063 TI - Expressed sequences from the basidiomycetous tree pathogen Heterobasidion annosum during early infection of scots pine. AB - The pattern of gene expression of the basidiomycete Heterobasidion annosum, causal agent of the root rot of conifers, was analysed during its interaction with pine roots. A complementary DNA (cDNA) library was constructed from total RNA extracted from H. annosum mycelia challenged with Scots pine seedling roots for 6 and 72h. Single pass sequencing of 1148 randomly selected cDNA clones resulted in 923 expressed sequence tags (ESTs). Contig analysis and sequence comparisons identified 318 unigene sequences, of which 62 were repeatedly sampled. A putative cellular function was assigned to 223 contigs (70%) that showed a moderate to high homology to protein sequences from public databases. Variations in expression levels during the infection process were monitored on a set of 96 unigenes by reverse northern using dot hybridisation. Seven unigenes (7%) were shown to be either up (4) or down (3) regulated during interaction of the fungus with pine roots. Fungal genes differentially expressed during contact with roots include genes encoding mitochondrial proteins, a cytochrome P450 and a vacuolar ATP synthase. PMID- 12742069 TI - Dantrolene inhibits NMDA-induced 45Ca uptake in cultured cerebellar granule neurons. AB - Dantrolene is an inhibitor of a skeletal muscle subtype of ryanodine receptors that stabilizes intracellular calcium concentrations and exerts neuroprotective effects in neurons submitted to excitotoxic challenges. The mechanisms of dantrolene-induced neuroprotection are not clear. In this study, using a model of cultured rat cerebellar granule neurons, we demonstrated that dantrolene inhibits NMDA-evoked 45Ca uptake, indicating that this drug may inhibit the activity of NMDA receptor channels. Primary neuronal cultures were incubated for 10 min in Mg(2+)-free ionic medium with NMDA and 45Ca in the presence of different concentrations of dantrolene, then radioactivity in neurons was measured by liquid scintillation spectroscopy. The results demonstrated that dantrolene, applied at micromolar concentrations, inhibits NMDA-evoked 45Ca uptake in neurons in a dose-dependent manner. DMSO, a vehicle to dantrolene, in concentrations used in this study had no effect on NMDA-evoked 45Ca uptake. These results, indicating that dantrolene inhibits activation of the NMDA receptors, might at least partially explain the mechanisms of a dantrolene-evoked protection of neurons against excitotoxicity mediated by agonists of NMDA receptors. PMID- 12742070 TI - Glutamine transport at the blood-brain and blood-cerebrospinal fluid barriers. AB - Glutamine has multiple physiological and pathophysiological roles in the brain. Because of their position at the interface between blood and brain, the cerebral capillaries and the choroid plexuses that form the blood-brain barriers (BBB) and blood-cerebrospinal fluid (CSF) barriers, have the potential to influence brain glutamine concentrations. Despite this, there has been a paucity of data on the mechanisms and polarity of glutamine transport at these barrier tissues. In situ brain perfusion in the rat, indicates that blood to brain L-[14C]glutamine transport at the blood-brain barrier is primarily mediated by a pH-dependent, Na(+)-dependent, System N transporter, but that blood to choroid plexus transport is primarily via a pH-independent System N transporter and a Na(+)-independent carrier that is not System L. Transport studies in isolated rat choroid plexuses and primary cultures of choroid plexus epithelial cells indicate that epithelial L-[14C]glutamine transport is polarized (apical uptake>basolateral) and that uptake at the apical membrane is mediated by pH dependent System N transporters (identified as SN1 and SN2 by polymerase chain reaction) and the Na(+) independent System L. Blood-brain barrier System N transport is markedly effected by cerebral ischemia and may be a good marker of endothelial cell dysfunction. The multiple glutamine transporters at the blood-brain and blood-CSF barriers may have role in meeting the metabolic needs of the brain and the barrier tissues themselves. However, it is likely that the main role of these transporters is removing glutamine, and thus nitrogen, from the brain. PMID- 12742071 TI - Asymmetry of glutamine transporters in cultured neural cells. AB - Transfer of glutamine between astrocytes and neurons is an essential part of the glutamate-glutamine cycle in the brain. Transport of glutamine was investigated in primary cultures of astrocytes and neurons and compared to glutamine transport in cell lines with glial and neuronal properties. Glutamine uptake in astrocytes was mainly mediated by general amino acid transporters with properties similar to ASCT2, LAT1, LAT2, SN1 and y(+)LAT2. In cultured neurons, transport activities were detected consistent with the presence of LAT1, LAT2 and y(+)LAT2, but the most prominent activity was a novel Na(+)-dependent glutamine transporter that could be inhibited by D-aspartate. The mRNA for system A isoforms ATA1 and ATA2 was detected in both neurons and astrocytes, but system A activity was only detected in neurons. ASCT2 on the other hand appeared to be astrocyte-specific. The cell lines F98 and 108CC-15, having astroglial and neuronal properties, respectively, expressed sets of glutamine transporters that were unrelated to those of the corresponding primary culture and are thus of limited use as models to study transfer of glutamine between astrocytes and neurons. PMID- 12742072 TI - Excitotoxic mechanism of cell swelling in rat cerebral cortical slices treated acutely with ammonia. AB - Swelling of CNS cells due to endogenous ammonia is a major cause of cerebral oedema in hyperammonaemic encephalopathies. In the present study, incubation in the presence of 5mM ammonium acetate ("ammonia") decreased steady-state distribution of [14C]inulin within incubated rat cerebrocortical minislices, indicating cell swelling. NMDA receptor antagonists, MK-801 ((+)-5-methyl-10,11 dihydro-5H-dibenzo [a,d]cycloheptene-5,10-imine maleate,10 microM) and DL-AP-5 (DL-2-amino-5-phosphonovaleric acid, 250 microM), a nitric oxide synthase inhibitor, L-nitroarginine (L-NNA, 500 microM), and an antioxidant, taurine (Tau, 10 mM), markedly attenuated the cell volume-increasing effect of ammonia. The effect of Tau (10mM) was abolished by the GABA(A) receptor antagonist bicuculline (100 microM), but was unaffected by the Tau transport inhibitor guanidynoethyl sulfonate (GES, 500 microM). Ammonia increased the slice content of Gln, an amino acid whose excess accumulation has been implicated in hyperammonemic oedema. However, treatments that reduced the cell volume did not affect Gln content. These results indicate that ammonia-induced cell swelling is in a large degree mediated by overactivation of NMDA receptors and the ensuing generation of NO and free radicals. PMID- 12742073 TI - Expression of monocarboxylic acid transporters (MCT) in brain cells. Implication for branched chain alpha-ketoacids transport in neurons. AB - The alpha-ketoisocaproic acid (KIC) is a short branched-chain monocarboxylate, which accumulates in neural cells. It plays an important role in maintaining nitrogen balance in the brain, a process of a great importance for shuttling of glutamine and glutamate between astrocytes and neurons. Higher accumulation of KIC in isolated cerebral cortex neurons at lower external pH, as well as sensitivity of this process to alpha-cyano-4-hydroxycinnamate indicate an involvement of a transporter, belonging to the family of monocarboxylate transporters (MCT).The expression of MCT1 and MCT2 isoforms in the brain cells was studied using reverse transcriptase-polymerase chain reaction (RT-PCR) method. The mRNA coding MCT1 was detected in astrocytes, brain endothelial cells, tumour cells (neuroblastoma and glioma) and in cortex neurons of newborn rats, but not in adult ones. MCT2, which is less abundant isoform than MCT1, was expressed in astrocytes, in brain endothelial cells and at low level in newborn rats' neurons, being absent in neurons from adult brain.The observed sensitivity of KIC accumulation towards SH-groups reagents did not fit to the known characteristics of MCT1 and MCT2. Therefore, the change of MCT expression during brain development, as well as lack of MCT1 and MCT2 in neurons of adults, point to another MCT isoform being involved in alpha-ketoisocaproic acid accumulation. This could be either one of other known MCT isoforms or a new member of family MCT, specific towards branched chain alpha-ketoacids. PMID- 12742074 TI - Differential roles of alanine in GABAergic and glutamatergic neurons. AB - Studies in different preparations of neurons and astrocytes of alanine transport and activities of its metabolizing enzyme alanine aminotransferase have led to the proposal that this amino acid is preferentially synthesized in astrocytes and transferred from the astrocytic to the neuronal compartment. From a functional point of view this may well be the case in a GABAergic synapse since theoretically alanine can be utilized as a metabolic fuel in GABAergic neurons where the GABA shunt is operating. Thus, a metabolic scheme is proposed, according to which alanine catabolism is coupled to the TCA cycle where the GABA shunt replaces the alpha-ketoglutarate dehydrogenase/succinyl CoA synthetase reactions. In a glutamatergic synapse in which the large demand for synthesis of neurotransmitter glutamate leads to a large production of ammonia, it is possible that alanine could play a completely different role. Hence, experimental evidence is reviewed suggesting that alanine may serve as a carrier of ammonia nitrogen from the neuronal compartment to the astrocytic compartment using a flux of lactate in the opposite direction to account for transfer of the C-3 carbon skeleton. PMID- 12742076 TI - Direct, noninvasive measurement of brain glycogen metabolism in humans. AB - The concentration and metabolism of the primary carbohydrate store in the brain, glycogen, is unknown in the conscious human brain. This study reports the first direct detection and measurement of glycogen metabolism in the human brain, which was achieved using localized 13C NMR spectroscopy. To enhance the NMR signal, the isotopic enrichment of the glucosyl moieties was increased by administration of 80 g of 99% enriched [1-13C]glucose in four subjects. 3 h after the start of the label administration, the 13C NMR signal of brain glycogen C1 was detected (0.36+/-0.07 micromol/g, mean+/-S.D., n=4). Based on the rate of 13C label incorporation into glycogen and the isotopic enrichment of plasma glucose, the flux through glycogen synthase was estimated at 0.17+/-0.05 micromol/(gh). This study establishes that brain glycogen can be measured in humans and indicates that its metabolism is very slow in the conscious human. The noninvasive detection of human brain glycogen opens the prospect of understanding the role and function of this important energy reserve under various physiological and pathophysiological conditions. PMID- 12742075 TI - In vivo 13C NMR assessment of brain glycogen concentration and turnover in the awake rat. AB - Brain glycogen metabolism was recently observed in vivo and found to be very slow in the lightly alpha-chloralose anesthetized rat [J. Neurochem. 73 (1999) 1300]. Based on that slow turnover, the total glycogen content in the awake rat brain and its turnover time were assessed after administering 13C-labeled glucose for 48 h. Label incorporation into glycogen, glucose, amino acid, and N-acetyl aspartate (NAA) resonances was observed. The amount of 13C label incorporated into glycogen was variable and did not correlate with that in glutamate (r=-0.1, P>0.86). However, the amount of 13C label incorporated into glycogen was very similar to that in NAA (r=0.93), implying similar turnover times between brain glycogen and NAA (approximately 10 h). Absolute quantification of the total concentration of brain glycogen in the awake, normoglycemic rat yielded 3.3+/-0.8 micromol/g (n=6, mean+/-S.D.). PMID- 12742077 TI - Lactate as a pivotal element in neuron-glia metabolic cooperation. AB - Lactate has been considered for a long time as a metabolic waste and/or a sign of hypoxia in the central nervous system. Nevertheless, clear evidence that lactate can constitute an adequate energy substrate for brain tissue has been provided as early as in the 1950s with the pioneering work of McIlwain in brain slices. Over the years, several studies using different approaches have confirmed that lactate is efficiently oxidized by brain cells in vitro. Moreover, lactate has been shown under certain circumstances to have a neuroprotective effect and support neuronal activity. Similar confirmation of lactate utilization in vivo as well as putative neuroprotection in various excitotoxic models has been provided. Lactate was even shown to restore cognitive performance upon an hypoglycemic episode in humans. More recently, it was proposed that lactate could be produced by astrocytes and released in the extracellular space to form a pool readily available for neurons in case of high energy demands. Several elements support the concept of a lactate shuttle between astrocytes and neurons in the central nervous system. Among them, the description of specific monocarboxylate transporters found on both astrocytes and neurons is an important observation consistent with this concept. Interestingly, lactate shuttles between different cell types within the same organ have been described outside the central nervous system, notably in muscle and testis. Thus, lactate is emerging as a valuable intercellular exchange molecule in different systems including the brain where it might be an essential element of neuron-glia metabolic interactions. PMID- 12742078 TI - Neighborly interactions of metabolically-activated astrocytes in vivo. AB - Metabolic responses of brain cells to a stimulus are governed, in part, by their enzymatic specialization and interrelationships with neighboring cells, and local shifts in functional metabolism during brain activation are likely to be influenced by the neurotransmitter system, subcellular compartmentation, and anatomical structure. Selected examples of functional activation illustrate the complexity of metabolic interactions in working brain and of interpretation of changes in brain lactate levels. The major focus of this article is the disproportionately higher metabolism of glucose compared to oxygen in normoxic brain, a phenomenon that occurs during activation in humans and animals. The glucose utilized in excess of oxygen is not fully explained by accumulation of glucose, lactate, or glycogen in brain or by lactate efflux from brain to blood. Thus, any lactate derived from the excess glucose could not have been stoichiometrically exported to and metabolized by neighboring neurons because oxygen consumption would have otherwise increased and matched that of glucose. Metabolic labeling of tricarboxylic acid cycle-derived amino acids increased during brief sensory stimulation, reflecting a rise in oxidative metabolism. Brain glycogen is mainly in astrocytes, and its level falls throughout the stimulus and early post-activation interval. Glycogenolysis cannot be accounted for by lactate accumulation or oxidation; there must be rapid product clearance. Glycogen restoration is slow and diversion of glucose from oxidative pathways for its re-synthesis could reduce the global O(2)/glucose uptake ratio; astrocytes could downshift this ratio for up to an hour after 5 min stimulus. Morphological studies of astrocytes reveal a paucity of cytoplasm and organelles in the fine processes that surround synapses and form gap junction connections with neighboring astrocytes. Specialized regions of astrocytes, e.g. their endfeet and thin peripheral lamellae, are likely to have compartmentalized metabolic activities. Anatomical constraints imposed upon the fine processes might require preferential utilization of glycolysis to satisfy their energy demands, but rapid lactate clearance would then be essential, since its accumulation would inhibit glycolysis. Gap junctional connections between neighboring astrocytes provide a mechanism for rapid metabolite spreading via the astrocytic syncytium and elimination of by-products. Local structure-function relationships need to be incorporated into experimental models of neuron-astrocyte and astrocyte-astrocyte interactions in working brain. PMID- 12742079 TI - Cataplerotic TCA cycle flux determined as glutamate-sustained oxygen consumption in primary cultures of astrocytes. AB - Utilization of glucose by adult brain as its metabolic substrate does not mean that glutamate cannot be synthesized from glucose and subsequently oxidatively degraded. Between 10 and 20% of total pyruvate metabolism in brain occurs as formation of oxaloacetate (OAA), a tricarboxylic acid (TCA) cycle intermediate, from pyruvate plus CO(2). This anaplerotic ('pool-filling') process occurs in astrocytes, which in contrast to neurons express pyruvate carboxylase (PC) activity. Equivalent amounts of pyruvate are converted to acetylcoenzyme A and condensed with oxaloacetate to form citrate (Cit), which is metabolized to alpha ketoglutarate (generating oxidatively-derived energy), glutamate and glutamine and transferred to neurons in the glutamate-glutamine cycle and used as precursor for transmitter glutamate. Since the blood-brain barrier is poorly permeable to glutamate and its metabolites, net synthesis of glutamate must be followed by degradation of equivalent amounts of glutamate, a cataplerotic ('pool-emptying') process, in which glutamate is converted in the TCA cycle to malate or oxaloacetate (generating additional energy), which exit the cycle to form one molecule pyruvate. To obtain an estimate of the rate of astrocytic oxidation of glutamate the rate of oxygen consumption was measured in primary cultures of mouse astrocytes metabolizing glutamate in the absence of other metabolic substrates. The observed rate is compatible with complete oxidative degradation of glutamate. PMID- 12742080 TI - Beta-amyloid and brain-derived neurotrophic factor, BDNF, up-regulate the expression of glutamate transporter GLT-1/EAAT2 via different signaling pathways utilizing transcription factor NF-kappaB. AB - Malfunctioning of high-affinity glutamate transporters is believed to contribute to the accumulation of toxic concentrations of glutamate and, thus, trigger the cellular mechanisms of neurodegeneration. Emerging data point to the presence of excitotoxic component in Alzheimer's disease (AD) and aberrant expression of glutamate transporters in this neurodegenerative malady. Neuronal soluble factors are essential for differential expression and fine tuning of the astroglial glutamate transporters, GLT-1/EAAT2 and GLAST/EAAT1. However, the nature of factors specifically affecting glutamate uptake in AD is largely unknown. The overproduction of neurotoxic beta-amyloid peptide (Abeta), a major constituent of amyloid plaques, and marked down-regulation of BDNF, a neuroprotective factor, are hallmarks of AD pathophysiology. None of these typically neuronal factors was capable of changing the pattern of glutamate transporter expression in undifferentiated rat astrocytes that predominantly expressed GLAST. In differentiated astrocytes, BDNF and, to a lesser extent, subtoxic concentrations of Abeta 1-42 (1-5 microM) induced the expression of GLT-1 and increased glutamate uptake, whereas the GLAST levels were unaltered by these factors. The BDNF-dependent up-regulation of GLT-1 in differentiated astrocytes was partially antagonized by the activation of metabotropic glutamate receptor 4 (mGluR4), but not by group I or II mGluRs. Activation of transcription factor NF-kappaB appeared to be a shared essential, but not a sufficient molecular event in the BDNF- or Abeta-dependent induction of GLT-1. The BDNF-dependent activation of NF kappaB and up-regulation of GLT-1 was critically dependent on the upstream activation of p42/p44 MAP kinase signaling, whereas the inhibition of these MAP kinases dramatically increased the Abeta-dependent activation of NF-kappaB and production of GLT-1. The capacity to up-regulate astroglial glutamate uptake system, that apparently represents a novel element in the neuroprotective repertoire of BDNF, can, however, provide adverse effect under certain insults when glutamate transporters start operating in reverse direction. The Abeta dependent up-regulation of GLT-1/EAAT2, more pronounced under the deficit of MAP kinase signaling, may attenuate synaptic efficacy and, thus contribute to the impairment of neuroplasticity in AD. PMID- 12742081 TI - Neurotoxic and neuroprotective effects of the glutamate transporter inhibitor DL threo-beta-benzyloxyaspartate (DL-TBOA) during physiological and ischemia-like conditions. AB - Maintenance of low extracellular glutamate ([Glu](O)) preventing excitotoxic cell death requires fast removal of glutamate from the synaptic cleft. This clearance is mainly provided by high affinity sodium-dependent glutamate transporters. These transporters can, however, also be reversed and release glutamate to the extracellular space in situations with energy failure. In this study the cellular localisation of the glutamate transporters GLAST and GLT-1 in organotypic hippocampal slice cultures was studied by immunofluorescence confocal microscopy, under normal culture conditions, and after a simulated ischemic insult, achieved by oxygen and glucose deprivation (OGD). In accordance with in vivo findings, GLAST and GLT-1 were primarily expressed by astrocytes under normal culture conditions, but after OGD some damaged neurons also expressed GLAST and GLT-1. The potential damaging effect of inhibition of the glutamate transporters by DL threo-beta-benzyloxyaspartate (DL-TBOA) was studied using cellular uptake of propidium iodide (PI) as a quantitative marker for the cell death. Addition of DL TBOA for 48 h was found to induce significant cell death in all hippocampal regions, with EC(50) values ranging from 38 to 48 microM for the different hippocampal subregions. The cell death was prevented by addition of the glutamate receptor antagonists NBQX and MK-801, together with an otherwise saturating concentration of DL-TBOA (100 microM). Finally, the effect of inhibition of glutamate release, via reverse operating transporters during OGD, was investigated. Addition of a sub-toxic (10 microM) dose of DL-TBOA during OGD, but not during the subsequent 48 h recovery period, significantly reduced the OGD induced PI uptake. It is concluded: (1) that the cellular expression of the glutamate transporters GLAST and GLT-1 in hippocampal slice cultures in general corresponds to the expression in vivo, (2) that inhibition of the glutamate transporters induces cell death in the slice cultures, and (3) that partial inhibition during simulation of ischemia by OGD protects against the induced PI uptake, most likely by blocking the reverse operating transporters otherwise triggered by the energy failure. PMID- 12742082 TI - GDNF pre-treatment aggravates neuronal cell loss in oxygen-glucose deprived hippocampal slice cultures: a possible effect of glutamate transporter up regulation. AB - Besides its neurotrophic and neuroprotective effects on dopaminergic neurons and spinal motoneurons, glial cell line-derived neurotrophic factor (GDNF) has potent neuroprotective effects in cerebral ischemia. The protective effect has so far been related to reduced activation of N-methyl-D-aspartate receptors (NMDAr). This study tested the effects of GDNF on glutamate transporter expression, with the hypothesis that modulation of glutamate transporter activity would affect the outcome of cerebral ischemia. Organotypic hippocampal slice cultures, derived from 1-week-old rats, were treated with 100 ng/ml GDNF for either 2 or 5 days, followed by Western blot analysis of NMDAr subunit 1 (NR1) and two glutamate transporter subtypes, GLAST and GLT-1. After 5-day exposure to GDNF, expression of GLAST and GLT-1 was up-regulated to 169 and 181% of control values, respectively, whereas NR1 was down-regulated to 64% of control. However, despite these changes that potentially would support neuronal resistance to excitotoxicity, the long-term treatment with GDNF was found to aggravate the neuronal damage induced by oxygen-glucose deprivation (OGD). The increased cell death, assessed by propidium iodide (PI) uptake, occurred not only among the most susceptible CA1 pyramidal cells, but also in CA3 and fascia dentata. Given that glutamate transporters are able to release glutamate by reversed action during energy failure, it is suggested that the observed increase in OGD-induced cell death in the GDNF-pretreated cultures was caused by the build-up of excitotoxic concentrations of extracellular glutamate released through the glutamate transporters, which were up-regulated by GDNF. Although the extent and consequences of glutamate release via reversal of GLAST and GLT-1 transporters seem to vary in different energy failure models, the present findings should be taken into account in clinical trials of GDNF. PMID- 12742083 TI - Glutamate uptake in blood platelets from epileptic patients. AB - Glutamate, the major excitatory amino acid neurotransmitter, is involved in epileptogenesis and initiation and spread of seizures. We studied glutamate uptake into blood platelets from patients with distinct epileptic syndromes: included were 20 patients with temporal lobe epilepsy and hippocampal sclerosis (TLE+HS), 20 with juvenile myoclonus epilepsy (JME) and 20 healthy volunteers matched for age and sex. The affinity of glutamate for the transporters was highest in patients with TLE+HS, but the maximal velocity of transport was highest in controls. There were no differences in the plasma levels of glutamate. Carbamazepine (CBZ), valproate (VPA) and lamotrigine (LTG) did not affect the uptake in vitro. The alterations observed in the uptake of glutamate in TLE+HS patients may reflect an up-regulated uptake of glutamate in the brain. PMID- 12742084 TI - Short- and long-term effects of MDMA ("ecstasy") on synaptosomal and vesicular uptake of neurotransmitters in vitro and ex vivo. AB - 3,4-Methylenedioxymethamphetamine (MDMA, "ecstasy") is a commonly abused drug which has been shown to be neurotoxic to serotonergic neurons in many species. The exact mechanism responsible for the neurotoxicity of MDMA is, however, poorly understood. In this study, the effects of MDMA on the synaptosomal and vesicular uptake of neurotransmitters were investigated. Our results show that MDMA (0.5-20 microM) reduces both synaptosomal and vesicular uptake of serotonin and dopamine in a dose dependent manner in vitro, while the uptake of glutamate and gamma aminobutyric acid (GABA) remains unaffected. Ex vivo experiments support the importance of the monoamines, with predominant dopaminergic inhibition at short term exposure (3 x 15 mg/kg; 2-h intervals), and exclusively serotonergic inhibition at long-term exposure (2 x 10 mg/kg per day; 4 days). This study also compares MDMA and the structurally related antidepressant paroxetine, in an attempt to reveal possible cellular mechanisms for the serotonergic toxicity of MDMA. One important difference between paroxetine and MDMA is that only MDMA has the capability of inhibiting vesicular uptake of monoamines at doses used. We suggest that inhibition of the vesicular monoamine transporter-2, and a following increase in cytoplasmatic monoamine concentrations, might be crucial for the neurotoxic effect of MDMA. PMID- 12742085 TI - Study of structure-function relationships in human glutamate dehydrogenases reveals novel molecular mechanisms for the regulation of the nerve tissue specific (GLUD2) isoenzyme. AB - In mammalian brain, glutamate dehydrogenase (GDH) is located predominantly in astrocytes, where is thought to play a role in transmitter glutamate's metabolism. Human GDH exists in GLUD1 (housekeeping) and GLUD2 (nerve tissue specific) isoforms, which share all but 15 out of their 505 amino acids. The GLUD1 GDH is potently inhibited by GTP, whereas the GLUD2 enzyme is resistant to this compound. On the other hand, the GLUD2 isoform assumes in the absence of GTP a conformational state associated with little catalytic activity, but it remains amenable to full activation by ADP and/or L-leucine. Site-directed mutagenesis of the GLUD1 gene at sites that differ from the corresponding residues of the GLUD2 gene showed that replacement of Gly456 by Ala made the enzyme resistant to GTP (IC(50)=2.8+/-0.15 microM) compared to the wild-type GDH (IC(50)=0.19+/-0.01 microM). In addition, substitution of Ser for Arg443 virtually abolished basal activity and rendered the enzyme dependent on ADP for its function. These properties may permit the neural enzyme to be recruited under conditions of low energy charge (high ADP:ATP ratio), similar to those that prevail in synaptic astrocytes during intense glutamatergic transmission. Hence, substitution of Ser for Arg443 and Ala for Gly456 are the main evolutionary changes that led to the adaptation of the GLUD2 GDH to the unique metabolic needs of the nerve tissue. PMID- 12742086 TI - Effect of methylmercury on glutamate metabolism in cerebellar astrocytes in culture. AB - The effect of methylmercury (MeHg) on [U-13C]glutamate metabolism was studied in cerebellar astrocytes using 13C nuclear magnetic resonance spectroscopy. The cells were preincubated in medium containing 25 or 50 microM MeHg and 10% fetal calf serum for 4h and then in medium with [U-13C]glutamate (0.5mM) for 2h. Labeled glutamate, glutamine and aspartate were observed both in the cell extracts and media, labeled glutathione in the cell extracts and labeled lactate and alanine in the media. The amount of glutamate removed from the media was decreased in the 50 microM MeHg group, furthermore, the levels of both labeled and unlabeled glutamine were decreased. This might indicate a decreased synthesis and/or increased degradation. An increase was observed for glutathione in the 25 microM group, which might be due to an upregulated synthesis of glutathione in response to the toxic effects of MeHg. The percentage of [U-13C]glutamate used for the synthesis of metabolites via the tricarboxylic acid cycle was increased in the presence of 50 microM MeHg. However, the percentage used for energy production was decreased in both groups, indicating selective mitochondrial vulnerability due to the inhibitory effect of MeHg. PMID- 12742087 TI - Characterization of depolarization-coupled release of glutamate from cultured mouse cerebellar granule cells using DL-threo-beta-benzyloxyaspartate (DL-TBOA) to distinguish between the vesicular and cytoplasmic pools. AB - Release of preloaded [3H]D-aspartate in response to depolarization induced by N methyl-D-aspartate (NMDA) or the endogenous agonist glutamate was characterized using cultured glutamatergic cerebellar granule neurons. Release from the vesicular and the cytoplasmic glutamate pools, respectively, was distinguished employing the competitive, non-transportable glutamate transport inhibitor DL threo-beta-benzyloxyaspartate (DL-TBOA). NMDA (300 microM)-induced release was enhanced (50%) by a simultaneous elevation of the extracellular potassium concentration to 15 mM, which lifts the voltage-dependent magnesium block of the NMDA receptors. This NMDA/K(+)-induced release was not sensitive to DL-TBOA (100 microM) but was inhibited by 75% in the presence of the unspecific calcium channel antagonist La(3+) (100 microM). Glutamate (100 microM) induced a large fractional release of the preloaded [3H]D-aspartate and in the presence of DL TBOA the release was reduced by approximately 50%. In contrast, release evoked by 25 microM glutamate was not inhibited by DL-TBOA. These results indicate that the release elicited by 100 microM glutamate is comprised of a significant glutamate transporter-mediated component in addition to the vesicular release while the NMDA/K(+)-induced release is vesicular in nature. It is likely that the high glutamate concentration (100 microM) may facilitate heteroexchange of the preloaded [3H]D-aspartate. PMID- 12742088 TI - Cyclic AMP-mediated regulation of striatal glutamate release: interactions of presynaptic ligand- and voltage-gated ion channels and G-protein-coupled receptors. AB - The presynaptic regulation of striatal glutamate transmission was investigated using D-[3H]aspartate and mouse striatal slices. Functional changes in voltage dependent and glutamate receptor-gated ion channels were elicited by pharmacologically modifying intracellular cyclic AMP formation via G-protein coupled receptor stimulation. The kainate (KA)-evoked release was potentiated by the stimulatory G-protein (G(s))-coupled beta-adrenoceptor agonist isoproterenol (ISO) in a concentration-dependent manner. This effect was mimicked by the specific calmodulin (CaM) antagonists trifluoperazine and calmidazolium. Tetrodotoxin (TTX), a blocker of Na(+) channels, did not affect the basal release but inhibited to the same degree the releases evoked by kainate alone and by kainate and isoproterenol together. Vinpocetine, a blocker of voltage-dependent Na(+) channels, did not alter the basal or the evoked release. The Na(+) channel activator veratridine enhanced the basal release in a concentration-dependent manner and isoproterenol attenuated this effect. The opposite effects of isoproterenol on the kainate- and veratridine-evoked releases may reflect prevention of the cyclic AMP-protein kinase A (PKA) phosphorylation cascade in striatal glutamatergic signal transduction. In addition, the calmidazolium induced potentiation of kainate-evoked release was thwarted by LY354740 and L-2 amino-4-phosphonobutanoate, agonists of the inhibitory G-protein (G(i))-coupled metabotropic group II and III glutamate receptors (mGluRs). Vinpocetine, which inhibits the CaM-dependent phosphodiesterase (PDE1), was likewise inhibitory. In turn, selective agonists and antagonists of the G(q)-protein-coupled group I mGluRs and (S)-3,5-dihydroxyphenylglycine (3,5-DHPG) and (RS)-1-aminoindan-1,5 dicarboxylate (AIDA), which modulate the intracellular Ca(2+) levels, did not alter the kainate-evoked release. The beta-adrenoceptor-mediated cyclic AMP accumulation seems to downregulate Na(+) channels but to enhance glutamate release by means of upregulation of kainate receptors. This regulation of presynaptic ligand- and voltage-gated ion channels is affected by the cAMP protein kinase A-dependent phosphorylation cascade and controlled by G(i)-protein coupled mGluRs. PMID- 12742089 TI - Effect of sodium lithium and proton concentrations on the electrophysiological properties of the four mouse GABA transporters expressed in Xenopus oocytes. AB - Mouse GABA transporters belong to the family of Na(+) and Cl(-) dependent neurotransmitter transporter. GABA transport, by these family members, was shown to be electrogenic and driven by sodium ions. It was demonstrated that, as in several other transporters, sodium binding and release by GAT1, GAT3 and BGT-1, the canine homolog of GAT2, resulted in the appearance of presteady-state currents. In this work we show that each of the four GABA transporters exhibit unique presteady-state currents when expressed in Xenopus oocytes. The properties of the presteady-state currents correspond to the transporters affinities to Na(+). At 100 mM GAT1 exhibited symmetric presteady-state currents at all imposed potentials, whereas GAT2 exhibited asymmetric presteady-state currents exclusively at negative imposed potentials, GAT3 or GAT4 exhibited presteady state currents predominantly at positive imposed potentials. GABA uptake by GAT2 and GAT4 was much more sensitive to external pH than GAT1 and GAT3. Reducing the external Na(+) concentration rendered the GABA uptake activity by GAT1 and GAT3 to be sensitive to pH. Lowering the external pH reduced the Na(+) affinity of GAT1. Substitution of the external Na(+) to Li(+) resulted in the appearance of leak currents exclusively at negative potentials in Xenopus oocyte expressing GAT1 and GAT3. Low Na(+) concentrations inhibited the leak currents of GAT1 but Na(+) had little effect on the leak currents of GAT3. Washing of occluded Na(+) in GAT1 enhanced the leak currents. Similarly addition of GABA in the presence of 80 mM Li(+), that presumably accelerated the release of the bound Na(+), also induced the leak currents. Conversely, addition of GABA to GAT3 expressing oocytes, in the presence of 80 mM Li(+), inhibited the leak currents. PMID- 12742090 TI - Effects of 3-hydroxy-4-amino-4,5,6,7-tetrahydro-1,2-benzisoxazol (exo-THPO) and its N-substituted analogs on GABA transport in cultured neurons and astrocytes and by the four cloned mouse GABA transporters. AB - The system of GABA transporters in neural cells constitutes an efficient mechanism for terminating inhibitory GABAergic neurotransmission. As such these transporter are important therapeutical targets in epilepsy and potentially other neurological diseases related to the GABA system. In this study a number of analogs of 3-hydroxy-4-amino-4,5,6,7-tetrahydro-1,2-benzisoxazol (exo-THPO), a promising lead structure for inhibitors of GABA uptake were investigated. It was found that the selectivity of N-acetyloxyethyl-exo-THPO for inhibition of the astroglial GABA uptake system was 10-fold as compared to inhibition of the neuronal GABA uptake system. Selectivity in this magnitude may provide potent anti-convulsant activity as has recently been demonstrated with the likewise glia selective GABA uptake inhibitor, N-methyl-exo-THPO. In contrast to the competitive inhibition of GABA uptake exhibited by N-substituted analogs of 4,5,6,7-tetrahydroisoxazolo[4,5-c]pyridin-3-ol (THPO), nipecotic acid, and guvacine, N-4,4-diphenyl-3-butenyl(DPB)-N-methyl-exo-THPO and 4-phenylbutyl-exo THPO exhibited non-competitive type inhibition kinetics. The lipophilic character of a number of GABA analogs was concluded by far to constitute the determining factor for the potency of these compounds as inhibitors of GAT1-mediated uptake of GABA. This finding underscores the complexity of the pharmacology of the GABA transport system, since these non-competitive inhibitors are structurally very similar to some competitive GABA uptake inhibitors. Whether these structure activity relationships for inhibition of GABA uptake may provide sufficient information for the development of new structural leads and to what extent these compounds may be efficient as therapeutical anti-convulsant agents remain to be elucidated. PMID- 12742091 TI - Characteristics of GABA release modified by glutamate receptors in mouse hippocampal slices. AB - The major part of hippocampal innervation is glutamatergic, regulated by inhibitory GABA-releasing interneurons. The modulation of [(3)H]GABA release by ionotropic and metabotropic glutamate receptors and by nitric oxide was here characterized in superfused mouse hippocampal slices. The ionotropic glutamate receptor agonists kainate, N-methyl-D-aspartate and 2-amino-3-hydroxy-5-methyl-4 isoxazolepropionate potentiated the basal GABA release. These effects were blocked by their respective antagonists 6-nitro-7-cyanoquinoxaline-2,3-dione (CNQX), dizocilpine and 2,3-dioxo-6-nitro-1,2,3,4-tetrahydrobenzo(f)quinoxaline-7 sulfonamide (NBQX), indicating receptor-mediated mechanisms. The NO-generating compounds S-nitroso-N-acetylpenicillamine (SNAP), sodiumnitroprusside and hydroxylamine enhanced the basal GABA release. Particularly the sodiumnitroprusside-evoked release was attenuated by the NO synthase inhibitor N(G)-nitro-L-arginine (L-NNA) and the inhibitor of soluble guanylyl cyclase 1H (1,2,4)oxadiazolo(4,3a)quinoxalin-1-one (ODQ), indicating the involvement of the NO/cGMP pathway. This inference is corroborated by the enhancing effect of zaprinast, a phosphodiesterase inhibitor, which is known to increase cGMP levels. The K(+)-stimulated hippocampal GABA release was reduced by the groups I and III agonists of metabotropic glutamate receptors (+/-)-1-aminocyclopentane-trans-1,3 dicarboxylate (t-ACPD) and L-(+)-2-amino-4-phosphonobutyrate (L-AP4), which effects were abolished by their respective antagonists (RS)-1-aminoindan-1,5 dicarboxylate (AIDA) and (RS)-2-cyclopropyl-4-phosphonophenylglycine (CPPG), again indicating modification by receptor-mediated mechanisms. PMID- 12742092 TI - Characterization of modes of release of amino acids in the ischemic/reperfused rat cerebral cortex. AB - Brain extracellular levels of glutamate, aspartate, GABA and glycine increase rapidly following the onset of ischemia, remain at an elevated level during the ischemia, and then decline over 20-30 min following reperfusion. The elevated levels of the excitotoxic amino acids, glutamate and aspartate, are thought to contribute to ischemia-evoked neuronal injury and death. Calcium-evoked exocytotic release appears to account for the initial (1-2 min) efflux of neurotransmitter-type amino acids following the onset of ischemia, with non vesicular release responsible for much of the subsequent efflux of these and other amino acids, including taurine and phosphoethanolamine. Extracellular Ca(2+)-independent release is mediated, in part by Na(+)-dependent amino acid transporters in the plasma membrane operating in a reversed mode, and by the opening of swelling-induced chloride channels, which allow the passage of amino acids down their concentration gradients. Experiments on cultured neurons and astrocytes have suggested that it is the astrocytes which make the primary contribution to this amino acid efflux. Inhibition of phospholipase A(2) attenuates ischemia-evoked release of both amino and free fatty acids from the rat cerebral cortex indicating that this group of enzymes is involved in amino acid efflux, and also accounting for the consistent ischemia-evoked release of phosphoethanolamine. It is, therefore, possible that disruption of membrane integrity by phospholipases plays a role in amino acid release. Recovery of amino acid levels to preischemic levels requires their uptake by high affinity Na(+) dependent transporters, operating in their normal mode, following restoration of energy metabolism, cell resting potentials and ionic gradients. PMID- 12742093 TI - Metabotropic glutamate receptors (mGluRs) are involved in early phase of memory formation: possible role of modulation of glutamate release. AB - Metabotropic glutamate receptors (mGluRs) groups I and II are involved in the cellular processes of long-term potentiation (LTP) and learning and memory formation. I.c.v. injection of the mGluRs agonist 1-aminocyclopentane-1,3 dicarboxylic acid (ACPD) can impair memory formation in some types of learning task. The role of mGluRs in neurotransmitters release and production of second messengers has been suggested. The aim of the present study was to determine the effect of i.c.v. administration of the new potent mGluRs agonist ABHxD-I and compare its effect with that of ACPD. We studied the effect of both agonists on acquisition and memory for a one-trial passive avoidance learning task in day-old chicks and on the training related glutamate (Glu) release. ACPD or ABHxD-I (50 nmole per chick, i.c.v. injection) were administered at different times before or after training and chicks were tested at various times after training. Chicks injected with ABHxD-I 30 min before training showed amnesia when tested 30 min or 3h after training. The amnestic effect of ACPD was significant only 30 min after training. Glu release evoked by 70 mM KCl was measured in slices prepared from the IMHV of chick brain isolated from animals injected with either ACPD or ABHxD I 30 min before training and tested 30 min after training. Glu concentration was measured using HPLC. Both ACPD and ABHxD-I significantly increased Glu release in slices isolated from untrained chicks (30 and 48% compare to control, respectively, P<0.05). Training itself increased Glu release (41% compared to control, P<0.01) and no additional effect of either ACPD or ABHxD-I was observed. These results suggest that mGluRs groups I and II are involved in the early stages of memory formation and that application of either of the studied mGluRs agonists may interfere with that process. The amnestic effect of ABHxD-I seems to be stronger and longer lasting. Although the mechanism of this effect still remains unclear, our results suggest that disregulation of Glu release by mGluR agonists may participate in this process. PMID- 12742094 TI - Manganese neurotoxicity and glutamate-GABA interaction. AB - Brain extracellular concentrations of amino acids (e.g. aspartate, glutamate, taurine) and divalent metals (e.g. zinc, copper, manganese) are primarily regulated by astrocytes. Adequate glutamate homeostasis is essential for the normal functioning of the central nervous system (CNS). Glutamate is of central importance for nitrogen metabolism and, along with aspartate, is the primary mediator of the excitatory pathways in the brain. Similarly, the maintenance of proper manganese levels is important for normal brain functioning. Several in vivo and in vitro studies have linked increased manganese concentrations with alterations in the content and metabolism of neurotransmitters, namely dopamine, gamma-aminobutyric acid, and glutamate. It has been reported by our laboratory and others, that cultured rat primary astrocytes exposed to manganese displayed decreased glutamate uptake, thereby increasing the excitotoxic potential of glutamate. Furthermore, decreased uptake of glutamate has been associated with decreased gene expression of glutamate:aspartate transporter (GLAST) in manganese exposed astroctyes. Additional studies have suggested that attenuation of astrocytic glutamate uptake by manganese may be a consequence of reactive oxygen species (ROS) generation. Collectively, these data suggest that excitotoxicity may occur due to manganese-induced altered glutamate metabolism, representing a proximate mechanism for manganese-induced neurotoxicity. PMID- 12742095 TI - Role of group I metabotropic glutamate receptors and NMDA receptors in homocysteine-evoked acute neurodegeneration of cultured cerebellar granule neurones. AB - Hyperhomocysteinemia is a risk factor in neurodegeneration. It has been suggested that apart from disturbances in methylation processes, the mechanisms of this effect may include excitotoxicity mediated by the N-methyl-D-aspartate (NMDA) receptors. In this study we demonstrate that apart from NMDA receptors, also group I metabotropic glutamate receptors participate in acute homocysteine (Hcy) induced neurotoxicity in cultured rat cerebellar granule neurones. Primary neuronal cultures were incubated for 30 min in the Mg(2+)-free ionic medium containing homocysteine and other ligands, and neurodegenerative changes were assessed 24h later using propidium iodide staining. D,L-Homocysteine given alone appeared to be a weak neurotoxin, with EC(50) of 17.4mM, whereas EC(50) for L glutamate was 0.17 mM. Addition of 50 microM glycine enhanced homocysteine neurotoxicity, and only that portion of neurotoxicity was abolished by 0.5 microM MK-801, an uncompetitive NMDA receptor antagonist. The net stimulation of 45Ca uptake by granule cells incubated in the presence of 25 mM D,L-homocysteine with 50 microM glycine was only 3% of the net uptake evoked by 1mM glutamate. Application of an antagonist of group I metabotropic glutamate receptors (mGluRs) LY367385 at 25 and 250 microM concentrations, induced a dose-dependent partial neuroprotection, whereas given together with MK-801 completely prevented neurotoxicity. In the absence of glycine, LY367385 and MK-801 given alone failed to induce neuroprotection, while applied together completely prevented homocysteine neurotoxicity. Agonist of group I mGluRs, 10 trans-azetidine-2,3 dicarboxylic acid (t-ADA) induced significant neurotoxicity. This study shows for the first time that acute homocysteine-induced neurotoxicity is mediated both by group I mGluRs and NMDA receptors, and is not accompanied by massive influx of extracellular Ca(2+) to neurones. PMID- 12742096 TI - Glutamine synthetase activity and glutamine content in brain: modulation by NMDA receptors and nitric oxide. AB - Acute intoxication with large doses of ammonia leads to rapid death. The main mechanism for ammonia elimination in brain is its reaction with glutamate to form glutamine. This reaction is catalyzed by glutamine synthetase and consumes ATP. In the course of studies on the molecular mechanism of acute ammonia toxicity, we have found that glutamine synthetase activity and glutamine content in brain are modulated by NMDA receptors and nitric oxide. The main findings can be summarized as follows. Blocking NMDA receptors prevents ammonia-induced depletion of brain ATP and death of rats but not the increase in brain glutamine, indicating that ammonia toxicity is not due to increased activity of glutamine synthetase or formation of glutamine but to excessive activation of NMDA receptors. Blocking NMDA receptors in vivo increases glutamine synthetase activity and glutamine content in brain, indicating that tonic activation of NMDA receptors maintains a tonic inhibition of glutamine synthetase. Blocking NMDA receptors in vivo increases the activity of glutamine synthetase assayed in vitro, indicating that increased activity is due to a covalent modification of the enzyme. Nitric oxide inhibits glutamine synthetase, indicating that the covalent modification that inhibits glutamine synthetase is a nitrosylation or a nitration.Inhibition of nitric oxide synthase increases the activity of glutamine synthetase, indicating that the covalent modification is reversible and it must be an enzyme that denitrosylate or denitrate glutamine synthetase.NMDA mediated activation of nitric oxide synthase is responsible only for part of the tonic inhibition of glutamine synthetase. Other sources of nitric oxide are also contributing to this tonic inhibition. Glutamine synthetase is not working at maximum rate in brain and its activity may be increased pharmacologically by manipulating NMDA receptors or nitric oxide content. This may be useful, for example, to increase ammonia detoxification in brain in hyperammonemic situations. PMID- 12742097 TI - Glutamine transport in C6 glioma cells shows ASCT2 system characteristics. AB - Previous studies from this laboratory have shown that glutamine (Gln) uptake in a rat astrocytoma-derived C6 cell line shows characteristics similar with the uptake of a model ASC system substrate, threonine, whose pH-dependence and partial tolerance of Li(+) substitution for Na(+) resemble the ASCT2 variant of the system. In support of the previous findings, RT-PCR analysis revealed that C6 cells strongly express ASCT2 mRNA, but not at all GlnT mRNA or NAT2 mRNA, the A and N system variants specifically engaged in Gln transport in normal CNS. Other features of Gln transport in C6 cells indicating the involvement of ASCT2 system included its resistance to ouabain and stimulation of Gln efflux from the cells in the presence of excess Gln or cysteine (Cys), demonstrating that the system operates in the exchange mode. Replacement of NaCl in the incubation medium with isoosmotic sucrose did neither significantly affect the kinetics, nor any other major characteristics of Gln or Thr transport, including its pH-dependence, inhibition by ASCT system substrates or resistance to the model system A substrate-N-methylamino-isobutyric acid (MeAiB). PMID- 12742098 TI - Potential mechanisms of enhanced GABA-mediated inhibitory neurotransmission in liver failure. PMID- 12742099 TI - Induction of the mitochondrial permeability transition in cultured astrocytes by glutamine. AB - Ammonia is a toxin that has been strongly implicated in the pathogenesis of hepatic encephalopathy (HE), and astrocytes appear to be the principal target of ammonia toxicity. Glutamine, a byproduct of ammonia metabolism, has been implicated in some of the deleterious effects of ammonia on the CNS. We have recently shown that ammonia induces the mitochondrial permeability transition (MPT) in cultured astrocytes, but not in neurons. We therefore determined whether glutamine is also capable of inducing the MPT in cultured astrocytes. Astrocytes were treated with glutamine (4.5 mM) for various time periods and the MPT was assessed by changes in 2-deoxyglucose (2-DG) mitochondrial permeability, calcein fluorescence assay, and by changes in cyclosporin A (CsA)-sensitive inner mitochondrial membrane potential (deltapsi(m)) using the potentiometric dye, JC 1. Astrocytes treated with glutamine significantly increased 2-DG permeability (120%, P<0.01), decreased mitochondrial calcein fluorescence, and concomitantly dissipated the deltapsi(m). All of these effects were blocked by CsA. These data indicate that glutamine induces the MPT in cultured astrocytes. The induction of the MPT by glutamine in astrocytes, and the subsequent development of mitochondrial dysfunction, may partially explain the deleterious affects of glutamine on the CNS in the setting of hyperammonemia. PMID- 12742100 TI - Cell-selective effects of ammonia on glutamate transporter and receptor function in the mammalian brain. AB - Increased brain ammonia concentrations are a hallmark feature of several neurological disorders including congenital urea cycle disorders, Reye's syndrome and hepatic encephalopathy (HE) associated with liver failure. Over the last decade, increasing evidence suggests that hyperammonemia leads to alterations in the glutamatergic neurotransmitter system. Studies utilizing in vivo and in vitro models of hyperammonemia reveal significant changes in brain glutamate levels, glutamate uptake and glutamate receptor function. Extracellular brain glutamate levels are consistently increased in rat models of acute liver failure. Furthermore, glutamate transport studies in both cultured neurons and astrocytes demonstrate a significant suppression in the high affinity uptake of glutamate following exposure to ammonia. Reductions in NMDA and non-NMDA glutamate receptor sites in animal models of acute liver failure suggest a compensatory decrease in receptor levels in the wake of rising extracellular levels of glutamate. Ammonia exposure also has significant effects on metabotropic glutamate receptor activation with implications, although less clear, that may relate to the brain edema and seizures associated with clinical hyperammonemic pathologies. Therapeutic measures aimed at these targets could result in effective measures for the prevention of CNS consequences in hyperammonemic syndromes. PMID- 12742101 TI - The effect of brominated flame retardants on neurotransmitter uptake into rat brain synaptosomes and vesicles. AB - The environmental levels of brominated flame retardants (BFRs) are increasing, but little is known about their toxic effects. In this paper, we show that some of the most important BFRs in commercial use today, have a neurotoxicological potential. Hexabromocyclododecane (HBCD) and tetrabromobisphenol-A (TBBPA) inhibit plasma membrane uptake of the neurotransmitters dopamine, glutamate and gamma-amino-n-butyric acid (GABA) at a concentration level similar to what previously found for polychlorinated biphenyls (PCBs) and even for ecstasy. The IC(50) value for HBCD on dopamine uptake was 4 microM, and the IC(50) values for TBBPA were 9, 6 and 16 microM for dopamine, glutamate and GABA, respectively. HBCD also inhibited glutamate uptake at low concentrations, but never achieved more than 50% inhibition. The inhibition was primarily due to their effect on the membrane potential, measured by the membrane potential marker tetraphenylphosphonium bromide (TPP(+)). Other brominated flame retardants such as octaBDE and decaBDE did not have any effects on uptake. TBBPA, HBCD and even the pentabrominated diphenylether mixture (pentaBDE, DE-71, Great Lakes) also inhibited the vesicular uptake of dopamine with an IC(50) value of 3, 3 and 8 microM, respectively. The neurotoxicological consequences of these findings for environmental contaminants such as BFRs and PCBs are discussed. PMID- 12742102 TI - Attentional selection of superimposed surfaces cannot be explained by modulation of the gain of color channels. AB - When two differently colored, superimposed patterns of dots rotate in opposite directions, this yields the percept of two superimposed transparent surfaces. If observers are cued to attend to one set of dots, they are impaired in making judgments about the other set. Since the two sets of dots are overlapping, the cueing effect cannot be explained by spatial attention. This has led to the interpretation that the impairment reflects surface-based attentional selection. However, recent single-unit recording studies in monkeys have found that attention can modulate the gain of neurons tuned for features such as color. Thus, rather than reflecting the selection of a surface, the behavioral effects might simply reflect a reduction in the gain of color channels selective for the color of the uncued set of dots (feature-based attention), as if viewing the surfaces through a colored filter. If so, then the impairment should be eliminated when the two surfaces are made the same color. Instead, we find that the impairment persists with no reduction in strength. Our findings thus rule out the color gain explanation. PMID- 12742103 TI - Oblique stimuli are seen best (not worst!) in naturalistic broad-band stimuli: a horizontal effect. AB - People with normal eyesight typically see horizontal and vertical gratings better than oblique gratings (Psychological Bulletin 78 (1972) 266; Perception 9 (1980) 37). In the present study we investigated whether this oblique effect anisotropy is still observed when viewing more complex visual stimuli that better correspond to the content encountered in everyday viewing of the world. We show that the ability to see oriented structure in an image consisting of broadband spatial content is indeed anisotropic, but that the pattern of this orientation bias is completely different from that obtained with simpler stimuli. Horizontal stimuli are seen worst and oblique stimuli are seen best when tested with more realistic broadband stimuli. We suggest that this "horizontal effect" would be useful in an evolutionary capacity by serving to discount the horizon and other oriented content that tends to dominate natural scenes and thereby increase the salience of objects contained in typical outdoor scenes. PMID- 12742104 TI - The what and where in visual masking. AB - A metacontrast mask suppresses the visibility of, without influencing the reaction time (RT) to, the target. We investigated whether this dissociation results from a sensori-motor pathway immune to masking effects or from the characteristics of stimulus timing in mutually inhibitory sustained and transient channels. For target visibility, para- and metacontrast yielded the usual U shaped functions. Peak paracontrast occurred at stimulus onset asynchronies (SOAs) of -150 to -100 ms. RTs were relatively low for metacontrast and did not show a systematic change as a function of SOA. The RT contribution from contour masking was greatest at an SOA of -150 ms (paracontrast) and declined to near zero in the metacontrast regime. The dissociation between visibility and RT seen in metacontrast did not occur in paracontrast, rejecting the theory that RTs are elicited by a single sensori-motor pathway immune to masking. The dependence of the dissociation on stimulus timing can be explained by RECOD, a dual-pathway model wherein fast and slow activities interact. PMID- 12742105 TI - Direct parameter specification of an attention shift: evidence from perceptual latency priming. AB - In the direct parameter specification (DPS) mode of sensorimotor control, response parameters can be specified by stimuli that are not consciously perceived [Psychological Research/Psychologische Forschung 52 (1990) 207]. DPS is contingent on the current intentions. The invisible stimuli can be processed for the purposes of sensorimotor control only if they match the actual intentions, for example, share task-relevant features. The present experiments explore whether attentional capture by masked abrupt-onset stimuli is mediated via DPS. Participants judged which of two visual targets appeared first. Masked primes preceded one of the targets. The primes were either similar to the targets or not, in shape, or in color. Target-like (task-relevant), but not distractor-like (task-irrelevant), primes facilitated perceptual latencies of targets trailing at their positions. Thus, the latency effects resulted from DPS of an attention shift, rather than from bottom-up capture or from top-down search for dynamic features. PMID- 12742106 TI - Task-specific perceptual learning on speed and direction discrimination. AB - Twenty-two nai;ve undergraduates participated in a psychophysical experiment designed to elucidate the neural events that allow us to see subtle motion differences. Half of the subjects practiced extensively on a direction discrimination task while the other half practiced extensively on a speed discrimination task. The stimulus conditions in the two groups were identical. The results indicated that the learning curves for direction discrimination were significantly steeper than those for speed discrimination. Additionally, the significant practice-based improvements on each motion task did not transfer to the other motion task. The different learning rates and the lack of transfer suggest that the neural events mediating speed discrimination are at least partially independent from those mediating direction discrimination, and vice versa, even under identical stimulus conditions. PMID- 12742107 TI - Orientation discrimination in foveal and extra-foveal vision: effects of stimulus bandwidth and contrast. AB - The parameter E2 is used in many spatial scaling studies to characterize the rate at which stimulus size must increase with eccentricity to achieve foveal levels of performance in detection and discrimination tasks. We examined whether the E2 for an orientation discrimination task was dependent on the spatial frequency bandwidth of the stimulus used. Two methods were employed. In Experiments 1 and 2 stimuli were presented at a fixed high level of contrast across viewing conditions. In both experiments the E2s recovered for narrowband stimuli were larger than those recovered for broadband stimuli. In Experiment 3 we controlled for the potentially confounding effects of perceptual contrast by measuring orientation thresholds over a range of stimulus contrast levels. Only thresholds which had reached an asymptotic level, such that increases in stimulus contrast led to no further changes to thresholds, were included in the calculation of E2. We observed that E2s recovered in the latter condition were in the range of 1.29 degrees -1.83 degrees and similar for narrowband and broadband stimuli. We conclude that a failure to consider the role of perceptual contrast may result in inflated estimates of E2. PMID- 12742109 TI - Ocular prevalence versus ocular dominance. AB - Ocular dominance manifests itself in tests that contain stereo-objects with a disparity beyond Panum's area, e.g. in pointing a finger. These tests force subjects to decide in favour of one or the other eye. In contrast, ocular prevalence is determined using stereo-targets imaged within Panum's areas. These tests allow a graded quantification of the balance between the eyes. Here we present the computer-based Freiburg Ocular Prevalence Test in which stereo disparate targets have to be aligned, and compare it with the Haase Stereo balance Test that requires an estimation of the horizontal distance between stationary stereo-disparate objects. In addition, we compare ocular prevalence with ocular dominance. METHODS: (1) We measured the influence of a neutral-grey filter in front of one eye to assess the suitability of the Freiburg and the Haase Tests in revealing graded amounts of ocular prevalence. (2) About 20 subjects with equal vision of their two eyes underwent the Freiburg and the Haase Tests for ocular prevalence, and Parson's Monoptoscope Test for ocular dominance. RESULTS: (1) In both the Freiburg and the Haase Tests, the neutral-grey filter shifted ocular prevalence by about 50%. (2) An ocular prevalence of more than 10% occurred in 13 of the 20 subjects using the Freiburg, and in 14 using the Haase Test. On average, the ocular prevalence was 24.1+/-3.8% in the Freiburg and 32.0+/-8.2% in the Haase Test. The dominant eye coincided with the prevalent eye in 15 of the 20 subjects. DISCUSSION: The effect of the neutral-grey filter indicated that both the Freiburg and the Haase Tests can be used to measure fractions of ocular prevalence, although the Freiburg Test carries a higher reproducibility. Spontaneous ocular prevalence occurs frequently in persons with equal vision of their two eyes. This suggests that ocular prevalence does not represent a condition that requires treatment. Rather, partial suppression of one eye, the correlate of ocular prevalence, may play a physiological role in that it helps to disregard double images at stereo-disparities close to the limits of Panum's area. PMID- 12742110 TI - The photopic ERG luminance-response function (photopic hill): method of analysis and clinical application. AB - With progressively brighter stimuli, the amplitude of the photopic b-wave first increases, briefly saturates and then decreases gradually to reach a plateau, where the amplitude of the b-wave equals that of the a-wave; a phenomenon previously presented as the photopic hill. The unique presentation of this luminance-response function seriously complicates its analysis with curve fitting equations such as that of Naka-Rushton used for scotopic electroretinogram. We report a method of analysis of the photopic hill based on easily identifiable and reproducible features of the ascending and descending limbs of this function. The clinical usefulness of these parameters is illustrated with selected cases of retinal disorders. PMID- 12742108 TI - Short-latency ocular following in humans is dependent on absolute (rather than relative) binocular disparity. AB - A previous study showed that the initial ocular following responses elicited by sudden motion of a large random-dot pattern were only modestly attenuated when that whole pattern was shifted out of the plane of fixation by altering its horizontal binocular disparity, but the same disparity applied to a restricted region of the dots had a much more powerful effect [Vision Research 41 (2001) 3371]. Thus, if the dots were partitioned into horizontal bands, for example, and alternate bands were moved in opposite directions to the left or right then ocular following was very weak, but if the (conditioning) dots moving in one direction were all shifted out of the plane of fixation (by applying horizontal disparity to them) then strong ocular following was now seen in the direction of motion of the (test) dots in the plane of fixation, i.e., moving images became much less effective when they were given binocular disparity. We sought to determine if the greater impact of disparity with the partitioned images was because there were additional relative disparity cues. We used a similar partitioned display and found that the dependence of ocular following on the absolute disparity of the conditioning stimulus had a Gaussian form with an x offset that was close to zero disparity and, importantly, this offset was almost unaffected by changing the absolute disparity of the test stimulus. We conclude from this that it is the absolute--rather than the relative--disparity that is important, and that ocular following has a strong preference for moving images whose absolute disparities are close to zero. This is consistent with the idea that ocular following selectively stabilizes the retinal images of objects in and around the plane of fixation and works in harmony with disparity vergence, which uses absolute disparity to bring objects of interest into the plane of fixation [Archives of Ophthalmology 55 (1956) 848]. PMID- 12742111 TI - Biological and environmental monitoring of hospital personnel exposed to antineoplastic agents: a review of analytical methods. AB - In order to assess occupational exposure of hospital personnel involved in the preparation and administration of antineoplastic drugs, biological and environmental monitoring are essential to identify the main exposure routes and to quantify potential health risks. If workplace contamination cannot be completely avoided, it is of utmost importance to reduce exposure to the lowest possible levels. To this aim, not only do education and training of the exposed subjects play an important role, but accurate standardized sampling techniques and analytical methods are also required. A critical overview of the most significant methods available in the literature is presented and their value is discussed, especially with respect to their sensitivity and specificity. In addition, attention is given to validation procedures and, consequently, to their reliability. The results from the most important surveys carried out at hospital departments are also discussed, with a view to improving both monitoring strategies and moreover working conditions. PMID- 12742112 TI - Highly sensitive quantification of vancomycin in plasma samples using liquid chromatography-tandem mass spectrometry and oral bioavailability in rats. AB - We developed a highly sensitive liquid chromatography-tandem mass spectrometry assay (LC-MS-MS) for a glycopeptide antibacterial drug, vancomycin (VCM), in rat plasma. After precipitating 100 micro l of plasma with 300 micro l of 10% trifluoroacetic acid-methanol (2:1, v/v), the supernatant was diluted with 300 micro l of distilled water and was passed through a filter. LC-MS-MS equipped with electrospray ionization in the positive ion mode used a pair of ions at 725/144 m/z for VCM in the multiple reaction-monitoring mode with a sample injection volume of 20 micro l. The calibration curve had a linear range from 0.01 to 20 micro g/ml when linear least square regression was applied to the concentration versus peak area plot. The drug in the sample was detected within 5 min. Precision, accuracy and limit of quantitation indicated that this method was suitable for the quantitative determination of VCM in rat plasma. Using this method, we defined for the first time that the oral bioavailability of VCM in rats was 0.069%. This method can be applied to basic pharmacokinetic and pharmaceutical studies in rats. PMID- 12742113 TI - Confirmatory analysis of 17beta-boldenone, 17alpha-boldenone and androsta-1,4 diene-3,17-dione in bovine urine by liquid chromatography-tandem mass spectrometry. AB - A sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS MS) method for confirmatory analysis of 17beta-boldenone (17beta-BOL), 17alpha boldenone (17alpha-BOL) and androsta-1,4-diene-3,17-dione (ADD) in bovine urine was developed. [2H(2)]17beta-Testosterone (17beta-T-d(2)) was used as the internal standard. Sample preparation involved enzymatic hydrolysis and purification on a C(18) solid-phase extraction column. Chromatographic separation of the analytes was obtained using an RP-C(18) HPLC column. LC-MS-MS detection was carried out with an atmospheric pressure chemical ionisation (APCI) source equipped with a heated nebulizer (HN) interface operating in the positive ion mode. For unambiguous hormone confirmation, three analyte precursor-product ion combinations were monitored during multiple-reaction monitoring (MRM) LC-MS-MS analysis. Overall recovery (%), repeatability (relative standard deviations, RSD, %) and within-laboratory reproducibility (RSD, %) ranged from 92.2 to 97.7%, from 6.50 to 2.94% and from 13.50 to 5.04%, respectively, for all analytes. The limit of quantification in bovine urine was 0.20 ng ml(-1) for 17beta-BOL and ADD and 0.50 ng ml(-1) for 17alpha-BOL. The validated method was successfully applied for determination of 17beta-BOL, 17alpha-BOL and ADD in a large number of bovine urine samples collected within the national Official Residue Control Program. PMID- 12742114 TI - Extractionless method for the determination of urinary caffeine metabolites using high-performance liquid chromatography coupled with tandem mass spectrometry. AB - Caffeine is metabolised in humans primarily by cytochromes P450 1A2 and 2A6, xanthine dehydrogenase/oxidase, and N-acetyltransferase 2. The activities of these enzymes show a large variation due to genetic polymorphisms and/or induction by xenobiotics. Ratios of different caffeine metabolites in urine or other body fluids are frequently used to characterise the individual/actual activity of these enzymes. The common analytical method involves extensive sample preparation, followed by HPLC-UV. The presence of numerous other UV-absorbing chemicals in body fluids affects the sensitivity and selectivity of this method. We have developed an HPLC-electrospray-MS-MS method for the determination of 11 caffeine metabolites and two internal standards after a simple, extractionless preparation. Blank urine, obtained after 5 days on a methylxanthine-free diet, contained small amounts of some caffeine metabolites, but no other components producing any confounding signals. Eleven metabolites and internal standards were recovered at 90 to 110% after addition to the blank urine (0.1 to 2.5 micro M in the final sample involving a 20-fold dilution of urine) in the 0.1-2.5 micro M concentration range. Other metabolites, 5-acetylamino-6-amino-3-methyluracil (AAMU) and 5-acetylamino-6-formylamino-3-methyluracil (AFMU), were detected with similar recovery and precision, but required higher concentrations (3 to 30 micro M). AFMU was completely converted into AAMU by a short alkalisation of urine. The method was explored in six healthy individuals after consuming coffee (4 mg caffeine per kg body mass). These experiments demonstrated the simplicity, high sensitivity and selectivity of the method under conditions used for phenotyping. PMID- 12742115 TI - Determination of a new atypical antipsychotic agent perospirone and its metabolite in human plasma by automated column-switching high-performance liquid chromatography. AB - A simple and sensitive column-switching high-performance liquid chromatographic (HPLC) method with fluorescence detection is described for the quantification of perospirone, a serotonin and dopamine antagonist, and its metabolite ID-15036 in human plasma. The test compounds were extracted from 2 ml of plasma using chloroform-hexane (30:70, v/v) and the extract was injected into a column I (TSK PW precolumn, 10 micro m, 35 x 4.6 mm I.D.) for clean-up and column II (C(18) STR ODS-II analytical column, 5 micro m, 150 x 4.6 mm I.D.) for separation. The peak was detected using a fluorescence detector set at Ex 315 nm and Em 405 nm, and the total time for a chromatographic separation was approximately 30 min. The method was validated for the concentration range from 0.1 to 100 ng/ml. Mean recoveries were 97% for perospirone and 96% for ID-15036. Intra- and inter-day relative standard deviations were less than 2.8 and 5.3% for perospirone and 2.4 and 4.4% for ID-15036, respectively, at the concentration range from 0.3 to 30 ng/ml. This method shows good specificity with respect to commonly prescribed psychotropic drugs, and it could be successfully applied for pharmacokinetic studies and therapeutic drug monitoring. PMID- 12742116 TI - Novel liquid chromatography-electrospray ionization mass spectrometry method for the quantification in human urine of microbial aromatic acid metabolites derived from dietary polyphenols. AB - An HPLC-ESI-MS-MS method was developed to quantify in human urine fourteen aromatic acids known as metabolites of dietary polyphenols. These metabolites were determined simultaneously in a single 20-min chromatographic analysis with multiple reaction monitoring detection. The inter- and intra-day precisions, calculated from quality control samples were 8.8 and 5.3%, respectively, and the mean accuracy was 2.3%. The method was tested on urine samples collected from one healthy volunteer who consumed a polyphenol-rich diet for 3 days. Increased levels of several aromatic acid metabolites were observed, demonstrating that the method can be used to detect changes in the excretion of microbial metabolites induced by the consumption of polyphenol-containing foods in humans. PMID- 12742117 TI - Determination of polycyclic aromatic hydrocarbons in milk samples by high performance liquid chromatography with fluorescence detection. AB - This paper describes a high-performance liquid chromatographic (HPLC) method for the determination of polycyclic aromatic hydrocarbons (PAHs) in milk samples. The method involves a liquid-liquid extraction procedure after saponification of milk samples with sodium hydroxide. Reproducible determination with highly sensitive detection was attained by HPLC with fluorescence detection using 1,2-bis(9 anthryl)ethane as an internal standard. The detection limits of 12 kinds of PAHs ranged from 1.3 to 76 ng/kg milk at a signal/noise ratio of 3. By the proposed method, the presence of 12 and 11 kinds of PAHs could be confirmed in commercial milk and human milk samples, respectively. The average concentrations of total PAHs (mean+/-SD, micro g/kg) were found to be 0.99+/-0.37 for commercial milk (n=14), 2.01+/-0.30 for infant formula (n=3) and 0.75+/-0.47 for human milk (n=51). High correlation coefficients between the concentrations of total PAHs and triglyceride were observed for commercial milk (r=0.659) and human milk (r=0.645). PMID- 12742118 TI - Properties of several glycidyl methacrylate-triallyl isocyanurate based affinity adsorbents for removing circulating immune complexes. AB - Biocompatible affinity adsorbents prepared from macroporous glycidyl methacrylate triallyl isocyanurate copolymer (GT) has been used for removing circulating immune complexes (CICs). In this work, adsorption of circulating immune complexes on GT-based affinity adsorbents has been studied by using batch and hemoperfusion studies. In batch mode, the equilibrium adsorption level was determined to be a function of the contact time, temperature, initial CIC concentration and adsorbent dosage. In animal hemoperfusion trials, removal of CICs is efficient and rapid. IgG, IgM and complement C3, C4 are minimally affected. There are negligible decreases in RBC, WBC, PLT and HB. Acid-base equilibrium, electrolytes and plasma proteins also are minimally affected. PMID- 12742119 TI - Liquid chromatography-tandem mass spectrometry assays for intracellular deoxyribonucleotide triphosphate competitors of nucleoside antiretrovirals. AB - This study was aimed to apply an LC-MS-MS method previously developed for intracellular nucleoside reverse transcriptase inhibitors-triphosphate (NRTI-TPs) to the determination of natural deoxyribonucleotides (dNTPs) in human peripheral blood mononuclear cells. The LC-MS-MS method was directly used in assay of dATP and dTTP. Interferences by ribonucleotides (rNTPs) prevented direct application to the two other analytes: dGTP and dCTP. A periodate oxidation procedure was therefore optimized to remove rNTPs from the cell medium in order to quantitate dCTP and dGTP. The determination of the intracellular ratio of NRTI-TP/dNTP in HIV-infected patients now involves use of the same chromatographic system for simultaneous assay of several analytes. PMID- 12742120 TI - Method development in high-performance liquid chromatography for high-throughput profiling and metabonomic studies of biofluid samples. AB - "Metabonomics" has in the past decade demonstrated enormous potential in furthering the understanding of, for example, disease processes, toxicological mechanisms, and biomarker discovery. The same principles can also provide a systematic and comprehensive approach to the study of food ingredient impact on consumer health. However, "metabonomic" methodology requires the development of rapid, advanced analytical tools to comprehensively profile biofluid metabolites within consumers. Until now, NMR spectroscopy has been used for this purpose almost exclusively. Chromatographic techniques and in particular HPLC, have not been exploited accordingly. The main drawbacks of chromatography are the long analysis time, instabilities in the sample fingerprint and the rigorous sample preparation required. This contribution addresses these problems in the quest to develop generic methods for high-throughput profiling using HPLC. After a careful optimization process, stable fingerprints of biofluid samples can be obtained using standard HPLC equipment. A method using a short monolithic column and a rapid gradient with a high flow-rate has been developed that allowed rapid and detailed profiling of larger numbers of urine samples. The method can be easily translated into a slow, shallow-gradient high-resolution method for identification of interesting peaks by LC-MS/NMR. A similar approach has been applied for cell culture media samples. Due to the much higher protein content of such samples non-porous polymer-based small particle columns yielded the best results. The study clearly shows that HPLC can be used in metabonomic fingerprinting studies. PMID- 12742121 TI - Simultaneous detection of trichloroethylene alcohol and acetate in rat urine by gas chromatography-mass spectrometry. AB - In order to better understand the cytotoxic effects of trichloroethylene (TCE) and its metabolites in TCE-induced carcinogenicity, it is necessary to determine the molecular species in biological samples. We have developed an efficient gas chromatography-mass spectrometry assay for the quantitative analysis of trichloroethylene alcohol and acetate. This method utilizes a simple esterification procedure, and a single liquid-liquid extraction with hexane dichloromethane (1:1) that allows >90% recovery of the metabolites, followed by gas chromatography-mass spectrometry. This protocol allows for the accurate, sensitive, and reproducible analysis of the toxic TCE metabolites. The utility of the assay is demonstrated through the analysis of TCE metabolites in urine from rats administered with TCE. The limit of quantitation (precision and accuracy<20%) was 1.7 ng/ml for TCE alcohol and 2.3 ng/ml for TCE acetate. PMID- 12742122 TI - Simple plasma work-up for a fast chromatographic analysis of homocysteine, cysteine, methionine and aromatic amino acids. AB - Simplified sample workup obviating protein precipitation and eluent evaporation commonly employed in earlier reports using chloroformate-mediated derivatization of aminothiols prior to mass spectrometric (MS) detection is presented. The reduction of disulfides in plasma is accomplished with dithiothreitol within minutes. A simultaneous derivatization with ethyl chloroformate (ECF) and extraction of derivatives into organic phase takes place within seconds. Along with S-amino acids, also aromatic amino acids can be determined during a 5-min run. Gas chromatography with flame ionization detection (GC-FID) proved to be sensitive enough to reach plasma homocysteine levels. A prerequisite for a reliable quantitation was fulfilled under the given conditions. Intra-assay precision was <5%, recoveries from spiked plasma complete (101.2%), detection and quantitation limits for homocysteine came to <1 and 3 micro mol/l. Our results were in full agreement with those obtained by liquid chromatography (r=0.999 for homocysteine and 0.987 for cysteine), and were close to two homocysteine immunoassays (r=0.991 and 0.939, respectively). PMID- 12742123 TI - Application of multiplexed capillary electrophoresis with laser-induced fluorescence (MCE-LIF) detection for the rapid measurement of endogenous extracellular signal-regulated protein kinase (ERK) levels in cell extracts. AB - Multiplexed (96-lane) capillary electrophoresis with laser-induced fluorescence (MCE-LIF) detection was used for the rapid analysis of extracellular signal regulated protein kinase (ERK) levels from in vitro cell extracts. The levels of ERK enzyme in cell extracts were determined by monitoring the conversion of a fluorescent-labeled peptide substrate to a phosphorylated fluorescent-labeled peptide product using MCE-LIF. The incorporation of a fluorescent internal standard was found to improve the precision of the analysis. The enzyme assay conditions including substrate concentration, reaction time and enzyme linear range were rapidly optimized using the MCE-LIF approach for both direct and immunoprecipitation-based ERK assays. The levels of ERK from in vitro cell extracts stimulated with angiopoietin 1 (Ang1*) were determined using the MCE-LIF approach. The advantages of MCE-LIF for developing and applying enzyme assays, as well as the figures of merit for the direct and immunoprecipitation ERK assays, are discussed. PMID- 12742124 TI - Development and validation of a liquid chromatographic-tandem mass spectrometric method for the determination of galantamine in human heparinised plasma. AB - Galantamine is an acetylcholinesterase inhibitor, recently approved for the treatment of mild-to-moderate Alzheimer's disease. To allow a higher throughput of samples, a new bioanalytical method for the determination of galantamine in human plasma was developed and validated. A stable isotope labelled internal standard was used. Sample preparation consisted of a simple one-step liquid liquid extraction with toluene. The extracts were analysed with positive ion TurboIonspray tandem mass spectrometry (LC-MS-MS). The method was validated in the 1-500-ng/ml range. The accuracy, precision, selectivity, lower limit of quantification, upper limit of quantification, linearity and extraction recovery were evaluated, as well as the stability of the compound in plasma, blood, methanol and 2% BSA solutions under different conditions. The method proved very rugged during the analysis of large numbers of samples from clinical trials. PMID- 12742125 TI - Validation of a method for quantification of ketobemidone in human plasma with liquid chromatography-tandem mass spectrometry. AB - A liquid chromatography tandem mass spectrometry (LC-MS-MS) method for determination of the analgesic aminophenol ketobemidone in human plasma is presented. Two preparation methods for plasma samples containing ketobemidone were compared, liquid-liquid extraction (LLE) and solid-phase extraction (SPE). Both methods showed good precision (n=10), 1.7% and 2.9%, respectively (0.04 micro M) and 1.1% and 2.5%, respectively (0.14 micro M). The accuracy was 98% and 103%, respectively (0.04 micro M) and 105% and 99%, respectively (0.14 micro M). Ketobemidone could be quantified at 0.43 nM, with a relative standard deviation of 17.5% (n=19) using LLE and 18.6% (n=10) using SPE. This level was an order of magnitude lower than earlier reported quantification limits. Quantitative data from plasma samples analyzed with LC-MS-MS were in good agreement with those obtained by gas chromatography with chemical ionization mass spectrometry (GC CI/MS). This indicates that LC-MS-MS is a good alternative method to GC-MS as it is more sensitive and time-consuming derivatization can be avoided. PMID- 12742126 TI - Simultaneous liquid chromatographic assessment of thiamine, thiamine monophosphate and thiamine diphosphate in human erythrocytes: a study on alcoholics. AB - An isocratic HPLC procedure for the assessment of thiamine (T), thiamine monophosphate (TMP) and thiamine diphosphate (TDP) in human erythrocytes is described. Several aspects of the procedure make it suitable for both clinical and research purposes: limits of detection and quantification of 1 and 2.5 nmol/l, respectively, recovery of 102% on average (range 93-112%), intra- and inter-day precisions within 5 and 9%, respectively, total elution time 15 min. This analytical methodology was applied to a case-control study on erythrocyte samples from 103 healthy subjects and 36 alcohol-dependent patients at risk of thiamine deficiency. Mean control values obtained were: T=89.6+/-22.7 nmol/l, TMP=4.4+/-6.6 nmol/l and TDP=222.23+/-56.3 nmol/l. T and TDP mean values of alcoholics were significantly lower than those of control cases: T=69.4+/-35.9 nmol/l (P<0.001) and TDP=127.4+/-62.5 nmol/l (P<10(-5)). The diagnostic role of TDP was evaluated and a significant role for thiamine was established in the study of alcohol related problems. PMID- 12742127 TI - Determination of polymyxin E1 in rat plasma by high-performance liquid chromatography. AB - A precise and accurate HPLC assay for polymyxin E(1) in rat and dog plasma has been validated. Samples and standards are extracted from plasma with a 96-well C(8) extraction disk plate. Sample extracts are derivatized with dansyl chloride, and polymyxin E(1) derivative is quantitated on a C(8) column by HPLC with fluorescence detection. The assay is linear in the range of 0.050-5.00 micro g/ml for polymyxin E(1). The precision and accuracy of polymyxin E(1) plasma assay was well within the recommended limits set in the FDA Guidance for Bioanalytical Method Validation. Polymyxin E(1) stability in rat and dog plasma for 24 h at room temperature and through three freeze-thaw cycles was demonstrated. PMID- 12742128 TI - Simple and sensitive determination of five quinolones in food by liquid chromatography with fluorescence detection. AB - A simple and sensitive high-performance liquid chromatographic (HPLC) method has been developed for the determination of five different quinolones: enrofloxacin, ciprofloxacin, sarafloxacin, oxolinic acid and flumequine in pork and salmon muscle. The method includes one extraction and clean-up step for the five quinolones together which are detected in two separated HPLC runs by means of their fluorescence. The proposed analytical method involves homogenizing of the tissue sample with 0.05 M phosphate buffer, pH 7.4 and clean-up by Discovery DS 18 cartridges. For chromatographic separation a Symmetry C(18) column is used in two different runs: (1) ciprofloxacin, enrofloxacin and sarafloxacin with acetonitrile-0.02 M phosphate buffer pH 3.0 (18:82) as mobile phase and the detector at excitation wavelength: 280 nm and emission wavelength 450 nm; and (2) oxolinic acid and flumequine with acetonitrile-0.02 M phosphate buffer pH 3.0 (34:66) as mobile phase and excitation wavelength: 312 nm and emission wavelength: 366 nm. Detection limit was as low as 5 ng g(-1), except for sarafloxacin which had a limit of 10 ng g(-1). Standard curves using blank muscle tissues spiked at different levels showed a good linear correlation coefficient, r(2) higher than 0.999 for all quinolones. PMID- 12742129 TI - Quantitative determination of free intracellular alpha-keto acids in neutrophils. AB - For the first time, a procedure is described for the quantitative analysis of free alpha-keto acid content in human neutrophils (PMNs) relative to single cell number by reversed-phase fluorescence high-performance liquid chromatography. The procedure is minimally invasive and is unsurpassed in the quality of PMN separation, ease of sample preparation as well as sample stability. This method can satisfy the rigorous demands for an ultra-sensitive, comprehensive and rapid intracellular alpha-keto acid analysis in particularly for the surveillance of severe diseases as well as cellular or organ dysfunction. PMID- 12742130 TI - Improved quantitative detection of 11 urinary phthalate metabolites in humans using liquid chromatography-atmospheric pressure chemical ionization tandem mass spectrometry. AB - Phthalates are widely used as industrial solvents and plasticizers, with global use exceeding four million tons per year. We improved our previously developed high-performance liquid chromatography-atmospheric pressure chemical ionization tandem mass spectrometric (HPLC-APCI-MS/MS) method to measure urinary phthalate metabolites by increasing the selectivity and the sensitivity by better resolving them from the solvent front, adding three more phthalate metabolites, monomethyl phthalate (mMP), mono-(2-ethyl-5-oxohexyl)phthalate (mEOHP) and mono-(2-ethyl-5 hydroxyhexyl)phthalate (mEHHP); increasing the sample throughput; and reducing the solvent usage. Furthermore, this improved method enabled us to analyze free un-conjugated mono-2-ethylhexyl phthalate (mEHP) by eliminating interferences derived from coelution of the glucuronide-bound, or conjugated form, of the mEHP on measurements of the free mEHP. This method for measuring phthalate metabolites in urine involves solid-phase extraction followed by reversed-phase HPLC-APCI MS/MS using isotope dilution with (13)C(4) internal standards. We further evaluated the ruggedness and the reliability of the method by comparing measurements made by multiple analysts at different extraction settings on multiple instruments. We observed mMP, monoethyl phthalate (mEP), mono-n-butyl phthalate (mBP), monobenzyl phthalate (mBzP), mEHP, mEHHP and mEOHP in the majority of urine specimens analyzed with DEHP-metabolites mEHHP and mEOHP present in significantly higher amounts than mEHP. PMID- 12742131 TI - Liquid chromatographic determination of carvedilol in human plasma. AB - A high-performance liquid chromatographic method for the quantitation of carvedilol in human plasma is presented. The method is based on protein precipitation with methanol, concentration of the supernatant by evaporation and reversed-phase chromatography with fluorimetric detection. The separation was performed on a Develosil 3 micro m ODS 100 x 4.6 mm I.D. column and the mobile phase consisted of acetonitrile-30 mM potassium dihydrogenphosphate buffer, pH 2 (30:70 v/v). With only 250 micro l of plasma used for sample preparation, the limit of quantitation 1.3 ng/ml was achieved. Dihydroergocristine mesylate was used as the internal standard. The between-day precision expressed by relative standard deviation was less than 6% and inaccuracy does not exceed 3%. The assay was used for pharmacokinetic studies. PMID- 12742132 TI - Amperometric detection of perphenazine at a carbon fiber micro-disk bundle electrode by capillary zone electrophoresis. AB - A simple method for determination of perphenazine by capillary zone electrophoresis with amperometric detection is described. The optimum conditions of separation and detection are 1.50 x 10(-3) mol/l Na(2)B(4)O(7)-1.0 x 10(-3) mol/l NaOH (pH 9.9) for the buffer solution, 18 kV for the separation voltage, 5 kV and 5 s for the injection voltage and the injection time, and 0.80 V versus saturated calomel electrode for the detection potential, respectively. The limit of detection is 5.0 x 10(-8) mol/l or 44 amol (S/N=3). The linear range of the calibration curve is 1.00 x 10(-7) to 1.00 x 10(-4) mol/l. The relative standard deviation is 1.5% for the migration time and 2.9% for the electrophoretic current at peak maximum. The method is applied to the determination of perphenazine in human urine. PMID- 12742133 TI - Determination of dialysate creatinine by micellar electrokinetic chromatography. AB - Micellar electrokinetic chromatography with UV absorbance detection has been applied for fast and selective determination of creatinine in samples of postdialysate fluid. Optimization of the method was performed, with the best results being obtained using a 30 mM borate-100 mM sodium dodecyl sulphate background electrolyte, pH 9, with the detector set at 235 nm and an applied voltage of 17 kV across a fused-silica capillary of 67 cm/75 micro m I.D. The linear range of the technique was over 2 orders of magnitude (5-1000 micro M). The developed analytical procedure is useful for the monitoring of clinical hemodialysis treatment, because creatinine levels in real undiluted samples of postdialysate range from 80 to 350 micro M. The separation system allows the analysis of about six to seven samples of spent dialysate per hour in almost real time. The determinations are not influenced by other components of dialysate fluid nor by other surrogates extracted from patient blood. The results of analysis using the developed procedure and the kinetic spectrophotometric Jaffe method conventionally used in clinical settings for creatinine determination are fully comparable. Successful clinical evaluation of the analytical system was performed. The developed system is useful for bloodless estimation of bioanalytical parameters of hemodialysis sessions such as creatinine-time profiles and total creatinine removal. Both these parameters are important in clinical models of hemodialysis therapy. PMID- 12742134 TI - Determination of glutathione in single human hepatocarcinoma cells by capillary electrophoresis with electrochemical detection. AB - A method for determination of glutathione (GSH) in single human hepatocarcinoma (HH) cells was described by capillary zone electrophoresis with electrochemical detection at a gold/mercury amalgam micro-disk electrode. When HH cells were washed with the running buffer instead of physiological buffer saline, only one electrophoretic peak for GSH is depicted on the electropherograms of single HH cells. When electroosmotic injection of 0.01 mol/l NaOH for lysing the cell introduced into the capillary, the lysis time can be shorten to 5 s. The whole cell injection and no need of derivatization reaction lead more accurate and precise results. The average amount of GSH in an individual HH cell is 22.3+/-5.8 fmol (mean+/-standard deviation), which is consistent with that of its homogenate. PMID- 12742135 TI - Capillary electrophoresis-based method to quantitate DNA-protein interactions. AB - A novel, rapid and simple capillary electrophoretic mobility shift assay (CEMSA) with laser-induced fluorescence (LIF) has been developed for the quantitative study of protein-DNA interactions. This method is particularly useful for the study of basic proteins, the most common of the DNA-interacting proteins. To avoid protein stickiness to the capillary walls we have introduced the use of neutral polyacrylamide that requires the use of reverse polarity. Under these conditions, excellent separation of DNA and protein-DNA complexes was obtained without the requirement of a gel matrix, thereby allowing the easy and reliable quantification of protein-DNA affinities. Analysis of the affinities of histones H2B and H4 for a synthetic oligo have been used to demonstrate the reproducibility and accuracy of this method. We have observed that H4 has a higher affinity for DNA than H2B, with half saturation fractions lying in the micromolar range. PMID- 12742136 TI - Analysis of diclofenac and its metabolites by high-performance liquid chromatography: relevance of CYP2C9 genotypes in diclofenac urinary metabolic ratios. AB - In humans, diclofenac is metabolised to 4'-hydroxy (OH), 3'-OH and 5-OH metabolites. The polymorphic CYP2C9 is involved in the metabolism of diclofenac to 4'-OH diclofenac and 3'-OH diclofenac. The aim of the present study was to develop a high-performance liquid chromatographic method to simultaneously measure diclofenac and its metabolites in urine, suitable for metabolic studies. After liquid-liquid extraction the compounds were separated in a reversed-phase column and measured by ultraviolet absorption at 282 nm. For all compounds intra day and inter-day variations were less than 7%, and the limits of quantitation were 0.25 mg/l. No analytical interference with endogenous compounds was found. The relationship between diclofenac metabolic ratios among different CYP2C9 genotypes is reported. The CYP2C9*3/*3 subject had the highest diclofenac/4'-OH ratios. However no difference was found between CYP2C9*2/*2 and *1/*1 genotypes. The chromatographic method developed was sensitive and reliable for the measurement of diclofenac and its metabolites simultaneously in human urine, and is suitable for use in diclofenac metabolism studies. PMID- 12742137 TI - Evidence-based medicine (EBM) in pediatric and adolescent gynecology. PMID- 12742138 TI - Multiple fibroids in a postmenopausal woman with Mayer Rokitansky Kuster Hauser syndrome. PMID- 12742139 TI - Prevalence and risk factors for LSD use among young women. AB - STUDY OBJECTIVE: To report the lifetime prevalence of lysergic acid diethylamide (LSD) and to identify unique correlates of using this substance in the last year among a large multiethnic sample of sexually active adolescent and young adult women aged 14 to 26 yrs. DESIGN, SETTING, PARTICIPANTS: A cross-sectional survey, administered at university-based ambulatory reproductive health clinics, was completed by 904 women between April and November of 1997 to identify risk factors associated with their use of LSD within the past 12 months. Subjects who reported lifetime, but not past 12 months', use of marijuana, LSD, or other illicit drugs were excluded, leaving a sample of 368 nonusers and 56 users of LSD. In addition, 231 young women who reported only using marijuana in the last year were used as a comparison group to identify unique factors associated with LSD use. RESULTS: Of the total sample (n=904), 13% (n=119) reported lifetime use of LSD, and 58% (n=536) reported lifetime use of marijuana. Logistic regression analyses controlling for age and race/ethnicity found distinct profiles for those who reported using LSD or only marijuana in the last year when compared to those who reported no substance use. Common to both groups was reporting being drunk at least 10 times during the last year, regular smoking of at least half a pack of cigarettes, and identification as a high-sexual-risk taker. However, LSD users as compared to nonusers were more likely to report white ethnicity (as compared to nonwhite), be less than or equal to 17 years of age (as compared to at least 18 years), report a history of physical abuse, and be categorized as having severe depressive symptomatology. In contrast, those who reported only using marijuana were more likely to report single marital status, young age at first intercourse, having half or more of their friends use marijuana, and poor grades. CONCLUSIONS: The female LSD user presents a distinct profile that might aid clinicians in identifying potential LSD use in this population as well as alerting clinicians to the relationship between LSD use and high-risk sexual behaviors. PMID- 12742140 TI - Predictors of repeat pregnancy in a program for pregnant teens. AB - STUDY OBJECTIVE: To describe repeat pregnancy among adolescents and to compare those who experienced a repeat pregnancy to those who did not. DESIGN, SETTING, PARTICIPANTS: Retrospective case control of all adolescents who entered and exited the Teen Mother & Child Program, a multidisciplinary clinic for pregnant and parenting teens and their children, between 1985 and 2000. MAIN OUTCOME MEASURE: Repeat pregnancy. RESULTS: Over the 16-yr study period, 1838 teens entered and exited the program with the mean time in the program of 1.9 yrs. 194 (10.6%) teens went on to have a second pregnancy, and 4 of those had a third pregnancy. Ninety-two percent (n=175) of these pregnancies resulted in a live birth. The mean interval between delivery of the initial child and delivery of the second child was 21.8 months (range 5.4-53.2). Teens repeating (R) were compared to those who did not repeat (NR). R were younger at entry and older at exit from the program (both P<0.0001). R were more likely to be Hispanic (P=0.009) or have a partner who was Hispanic (P=0.02). R were more likely to have experienced a poor initial pregnancy outcome (miscarriage, stillbirth) (P=0.03). R were more likely than NR to be in a stable relationships with the father of the baby (i.e., married or engaged) (P=0.03). Self-report of physical and sexual abuse, depression, and substance abuse were common, but did not differ between R and NR. However, R were more likely than NR to self-report suicide gestures/attempts and to have a significant psychiatric history (P=0.01, P=0.004). Only 24% of R and 26% of NR had completed high school by the time of exit from the program. CONCLUSIONS: Ten percent of adolescents served by comprehensive multidisciplinary teen pregnancy program experienced a repeat pregnancy while in the program. Compared to nonrepeaters, adolescents who experienced a repeat pregnancy were younger, were more frequently Hispanic, and were more likely to be in a stable relationship with the baby's father. Many of the program's clients have psychosocial factors reported in the literature to be associated with repeat pregnancy. A history of suicide gestures/attempts and a significant psychiatric history were more common in those who had a repeat pregnancy. Disappointingly, only about 25% of the adolescents completed high school by the time they exited the program whether they experienced a repeat pregnancy or not. Although directing interventions (e.g., mental health services, counseling those who miscarried) to teens who appear to be at highest risk for a repeat pregnancy may decrease their risk of repeating, all teens in our program would likely benefit from such services. PMID- 12742141 TI - Spasticity of the pelvic floor mimicking an obstructive anomaly. AB - STUDY OBJECTIVE: Hematocolpos or hydrocolpos in menstruating women raises suspicion of a partial uterine or vaginal obstruction. The study objective is to report two unusual cases of a spastic pelvic floor leading to urine collecting in the vagina and mimicking an outflow obstruction. DESIGN, SETTING, PARTICIPANTS: The study took place at a tertiary care university's Reproductive Health Care Clinic for Women with Developmental Disabilities and involved two patients with spastic quadriplegic cerebral palsy and developmental disabilities who presented with irregular menses and abdominal pain and whose radiological evaluations were suspicious for an outflow obstruction. Medical records, including clinic visits, radiological findings, and surgical findings, were reviewed. INTERVENTIONS: Both patients underwent ultrasound and MRI evaluation of their reproductive tracts that demonstrated fluid collections in the vagina. An examination under anesthesia was performed in one patient to rule out an obstruction. In the second patient, a pelvic examination under ultrasound observation revealed initial vaginal distension with urine, which resolved after placement of a speculum. RESULTS: In both cases, the pseudo-obstruction was felt to be a urine-distended vagina due to a spastic pelvic floor. CONCLUSION: A spastic pelvic floor in an incontinent patient with spastic quadriplegia may result in urine accumulation in the vagina, mimicking an outflow tract obstruction. If the history, pelvic examination, and radiological images are inconsistent, performing an ultrasound guided examination may assist with diagnosis. PMID- 12742142 TI - A spontaneous and uneventful pregnancy in a Turner mosaic with previous recurrent miscarriages. AB - BACKGROUND: Conception without ovum donation is very rare in patients with Turner syndrome, occurring mainly in mosaics, and only a third of these natural pregnancies are associated with normal outcome. CASE REPORT: A spontaneous pregnancy is described in a Turner mosaic (45,X/46,XX) with normal puberty and nine previous first-trimester miscarriages who had refused ovum donation. She also declined antenatal genetic diagnosis. Pregnancy remained uneventful with frequent surveillance and psychosocial support. She delivered a live boy weighing 3260 g (50(th) percentile) and measuring 48 cm (50(th) percentile) with normal karyotype at term. CONCLUSION: The diagnosis of mosaic Turner syndrome should be considered in women who have recurrent abortion. Pregnancy may have a good outcome in these patients despite the greater likelihood of recurrent miscarriage. PMID- 12742143 TI - Menstrual irregularity in female military cadets: comparison of data utilizing short-term and long-term recall. AB - STUDY OBJECTIVES: (1) To assess menstrual function in freshman cadets at a structured, rigorous military college utilizing a weekly e-mail questionnaire, (2) to compare these results to longer-term recall results from written questionnaires completed at the start and end of the same year, and (3) to determine if a relationship exists between personality attributes and menstrual regularity in this setting. DESIGN, SETTING, PARTICIPANTS: A questionnaire developed by the investigators about menstrual function was distributed weekly via e-mail and completed satisfactorily by 116 female freshman at the United States Military Academy (USMA), class of 1995. These data were compared to questionnaires completed by this same cohort at the start and end of the year, as well as to personality questionnaires distributed to some of these cadets as part of a larger study. Pearson's chi-square and analysis of variance were performed to determine statistical significance. RESULTS: Weekly e-mail data revealed that only 1.7% of cadets had regular menstrual periods throughout the whole year. The remaining 98.3% were irregular: 10.3% mildly, 35.3% moderately, 30.2% severely, and 22.4% extremely irregular. Only 0.8% of subjects did not menstruate at all during the year. In comparison, on long-term recall data, 90% reported some change in menstrual function, with 48% specifically reporting menstrual irregularity. Those with the greatest irregularity prior to USMA and those who expected greater irregularity at USMA reported the greatest irregularity during the year. Irregularity was associated with coping and with commitment to health on personality questionnaires. CONCLUSIONS: On data collected via weekly e-mail, almost all freshman females at USMA experienced menstrual irregularity. This data, utilizing short-term recall, yielded different, more dramatic results than the data utilizing long-term recall. A link between menstrual irregularity and both coping and commitment suggests that menstrual irregularity may be adaptive in this environment. PMID- 12742144 TI - Douching behaviors reported by adolescent and young adult women at high risk for sexually transmitted infections. AB - STUDY OBJECTIVE: To describe frequency of douching and reasons as timing to menses, vaginal symptoms, and coitus and the association of these behaviors to the diagnosis of three sexually acquired infections. DESIGN, SETTING, PARTICIPANTS: The study involved 160 females between the ages of 14 and 25 yrs attending a STD clinic and/or community adolescent health clinics. Subjects were eligible to enter the study if they had a positive test(s) for and/or were a contact of chlamydia, gonorrhea, trichomonas, and/or nongonococcal urethritis (NGU). Reevaluation for these infections occurred at the 1-month, 4-month, and 7 month visit with one-dose antibiotic treatment provided for positive tests. Data on douching was collected at the 7-month visit only. MAIN OUTCOME MEASURE: Results of tests for STI's using urine-based DNA-amplification techniques for chlamydia and gonorrhea and using self-obtained vaginal swabs for trichomonas culture. RESULTS: Nearly two-thirds (106/160) of the subjects ages 14-25 yrs completing the 7-month visit reported douching, with 67.7% (69/102) reporting douching once a month or more. Douching was more common in older, black participants, using injectable progestins for contraception. Douching was more common in those reporting more recent sexual partners. Douching related to menses was not associated with any of the three infections, while douching related to symptoms and coitus was associated with positive tests for infections. CONCLUSIONS: Results suggest that for this subset of teens at high risk for sexually acquired infections, douching is a commonly reported behavior. This study suggests that the linkage of douching and sexually acquired infections is associated with contraceptive choices, self-treatment of vaginal symptoms, and sexual risk behaviors but not menstrual hygiene. PMID- 12742145 TI - Physician knowledge and attitudes of Minnesota laws concerning adolescent health care. AB - STUDY OBJECTIVE: To determine physicians' knowledge and attitudes of medico-legal issues affecting adolescent health care. DESIGN, SETTING, PARTICIPANTS: A cross sectional, mailed survey was distributed to 900 randomly selected primary care physicians in Minnesota, and all eligible physicians in Olmsted County. INTERVENTIONS: Physicians were mailed a survey with questions concerning Minnesota consent and confidentiality laws. MAIN OUTCOME MEASURES: Physician knowledge and attitudes of consent and confidentiality laws. RESULTS: A total of 317 (26%) surveys were returned. Olmsted county physicians achieved a median score of 37.5% and non-Olmsted physicians achieved a median score of 50% correct on a test of knowledge. Using a scoring scale where -1 signified "a bad law," 0 signified "neither a good nor bad law," and +1 signified "a good law," attitude regarding the laws was a median of +0.5 for both groups. In all, 41.1% of Olmsted physicians and 51.7% of non-Olmsted physicians felt that the laws had affected them or their practice. Olmsted physicians were compared to a cohort of Olmsted parents previously reported, with statistically significant differences noted. CONCLUSIONS: Results suggest that primary care physicians are not knowledgeable of Minnesota laws affecting adolescent health care. Opinion of these laws was positive, with notable exceptions. Physicians lack a sense of impact of laws affecting adolescent health care. Physicians were more knowledgeable and felt more positively about the laws than did parents of adolescents. Lack of knowledge and the presence of certain attitudes identify areas where physicians could benefit from greater understanding. PMID- 12742146 TI - Metformin use in adolescent with PCOS--an invited editorial from the pediatric endocrinology perspective. PMID- 12742148 TI - Gynecological care for teenagers with disabilities. PMID- 12742147 TI - Teens, confidentiality, and HIPPA. PMID- 12742153 TI - Updates on the cytogenetics and molecular genetics of bone and soft tissue tumors: chondrosarcoma and other cartilaginous neoplasms. PMID- 12742154 TI - Blastoid mantle cell lymphoma: evidence for nonrandom cytogenetic abnormalities additional to t(11;14) and generation of a mouse model. AB - Mantle cell lymphoma (MCL) is characterized by the t(11;14)(q13;q32), which is associated with cyclin D1 hyperexpression and a poor prognosis. MCL cases have been shown to progress to a more aggressive disease but the molecular events responsible of this phenomenon have not been determined. We have established two cell lines from the pleural effusions of two patients with MCL that we have used for further cytogenetic characterization to better define the incidence and nature of secondary chromosome abnormalities using multicolor fluorescence in situ hybridization, whole chromosome paint, and specific probes. Both cell lines grew independently without growth factors. Using CCND1/IGH-specific probes, patient UPN1 was found to have a masked t(11;14). Numerous and complex chromosomal abnormalities were found in both cell lines affecting chromosomes 2, 8, 13, 18, 22, X, and Y. These abnormalities included 8p losses, suggesting the presence of an anti-oncogene in this region, rearrangements of 8q24, MYC gene, and translocations involving 8, X, and Y chromosomes, which might be significant in the pathogenesis of MCL progression. The use of the cell lines (UPN1) allowed us to generate a mouse model of human MCL, mimicking a disseminated lymphoma and leading to the death of the animals in 4 weeks. This blastoid MCL model could be of major interest to determine molecular events involved in MCL progression, allowing isolation of involved genes and their functional characterization, and to study the effects of new chemotherapy regimens in mouse models. PMID- 12742156 TI - Genomic rearrangements involving rDNA and centromeric heterochromatin in vulvar epidermoid carcinoma cell line A-431. AB - The cytogenetic and molecular cytogenetic characterization of the human cell line A-431 derived from a vulvar epidermoid carcinoma is presented. A combination of karyotyping, fluorescence in situ hybridization (FISH) with chromosome- and/or region-specific probes, M-FISH, RxFISH, and comparative genomic hybridization (CGH) analysis was used. Six marker chromosomes with rearrangements involving insertions of single or double nucleolar organizing regions (NORs) and/or homogeneously staining regions containing active and overexpressed NORs and regions of centromeric heterochromatin were found: der(6), der(7), der(17), der(21), dic(13;14), and dic(14;18). The chromosomal origin of 14 other marker chromosomes was elucidated. Amplification of the C-MYC oncogene at 8q24 was revealed in two marker chromosomes: dup(8)(q24) and der(15)t(8;15)(q22;p11). Confirming previous reports, amplification of the cyclin D1 gene within an abnormal chromosome 11, that is, der(11)t(7;11)(p15;q21), was also detected. Loss of the TP53 tumor suppressor gene was evidenced over two der(17). Good concordance was found among karyotyping, FISH analysis, and CGH. Although reasons for NOR amplification or ectopic location in the epidermal carcinoma A-431 cell line are not clear yet, our data suggest that these phenomena play a supporting role with regard to other amplified genes. Thus, the A-431 cell line would be an appropriate model to study the different mechanisms involved in human tumorigenesis. PMID- 12742155 TI - The dominant-negative effect of p53 mutants and p21 induction in tetraploid G1 arrest depends on the type of p53 mutation and the nature of the stimulus. AB - Many p53 mutant proteins possess a dominant-negative activity that is under the control of several factors, namely p53 mutations and the cell type. The goals of our study were to determine the following: (1) the dominant-negative effect of different p53 mutations in response to mitotic spindle inhibitors, and (2) if this dominant-negative activity is dependent on the nature of the stimulus. We therefore examined the cellular response of the near-diploid LoVo colon carcinoma cell line possessing two wild-type TP53 alleles and three other clones transfected with different TP53 mutants (p53-273H, p53-175H, and p53-143A) to treatments with different mitotic spindle inhibitors. Flow cytometric studies and analysis of retinoblastoma protein (pRb) dephosphorylation and 5-bromo-2' deoxyuridine incorporation by immunocytochemistry revealed a tetraploid G1 arrest of the wild-type LoVo clone and the p53-273H mutant clone after exposure to mitotic spindle inhibitors, preventing tetraploid cells from entering into an additional S phase. On the other hand, the p53-175H and p53-143A mutant clones re enter S phase with no apparent arrest. Therefore, our results confirm that p53 mutant dominant-negative activity and the tetraploid G1 arrest in response to mitotic spindle inhibitor treatment depend on the type of p53 mutation, involve p21 induction, and require pRb dephosphorylation. Moreover, when we compare our results with those obtained by other investigators after ionizing radiation exposure using the same cell lines, we identify the nature of the stimulus as a new factor that determines the dominant-negative effect of p53 mutants. PMID- 12742157 TI - Instability of chromosome structure in cancer cells increases exponentially with degrees of aneuploidy. AB - Structurally altered or marker chromosomes are the cytogenetic hallmarks of cancer cells, but their origins are still debated. Here we propose that aneuploidy, which is ubiquitous in cancer and inevitably unbalances thousands of synergistic genes, destabilizes the structure of chromosomes by catalyzing DNA breaks. Aneuploidy catalyzes such breaks by unbalancing teams of enzymes, which synthesize and maintain DNA and nucleotide pools, and even unbalancing histones via the corresponding genes. DNA breaks then initiate deletions, amplifications, and intra- and interchromosomal rearrangements. Our hypothesis predicts that the rate at which chromosomes are altered is proportional to the degree of aneuploidy: the more abnormal the number and balance of chromosomes, the higher the rate of structural alterations. To test this prediction, we have determined the rates at which clonal cultures of diploid and aneuploid Chinese hamster cells generate new, and thus nonclonal, structurally altered chromosomes per mitosis. Based on about 20 metaphases, the number of new, structurally altered chromosomes was 0 per diploid, 0-0.23 per near-diploid, 0.2-1.4 per hypotriploid, 3.25-4.8 per hypertriploid, and 0.4 per near-tetraploid cell. Thus, instability of chromosome structure increases exponentially with the deviation of ploidy from the normal diploid and tetraploid balances. The particular chromosomes engaged in aneuploidy also affected the rates of chromosome alteration, particularly at low aneuploidy indices. We conclude that aneuploidy is sufficient to cause structural instability of chromosomes. Further, we suggest that certain structurally altered chromosomes encode cancer-specific phenotypes that cannot be generated by unbalancing intact chromosomes. We also extend the evidence for aneuploidy causing numerical instability of chromosomes autocatalytically, and adduce evidence that aneuploidy can cause the many gene mutations of cancer cells that have been attributed to various mutator genes. PMID- 12742158 TI - Comprehensive cytogenetic analysis including multicolor spectral karyotyping and interphase fluorescence in situ hybridization in lymphoma diagnosis. a summary of 154 cases. AB - Cytogenetic analysis including multicolor spectral karyotyping (SKY) and interphase fluorescence in situ hybridization (FISH) was performed on 154 consecutive cases with suspected lymphoma. The cytogenetic results were reviewed in correlation with the final pathologic diagnosis. A diagnosis of lymphoma was established in 94 cases, with 16 Hodgkin lymphomas and 78 non-Hodgkin lymphomas (NHL). Cytogenetic results were obtained in 63 NHLs (81%); 61 of those showed abnormal karyotypes (97%). The t(14;18) or IGH-BCL2 fusion was detected in 83% (20/24) of follicular lymphomas and in 57% (12/21) of diffuse large B-cell lymphomas (DLBCL). The application of interphase FISH and SKY has contributed to a high detection rate of t(14;18) in DLBCLs. This study showed that genes at 1q25, 3p21, 3q21, 5q31, 6p23, 7q22, 8q11 approximately q12, 9q34, 11q23, 12q13, and 19q13.1 may have been involved as the less common changes in follicular lymphoma and DLBCL. Comparison of the recurrent secondary aberrations in the groups of follicular lymphoma and DLBCL revealed a pattern of clonal evolution from the changes rea(1)(p36), del(6q), +7, +12 or dup or trp(12)(q13q22), +der(18)t(14;18), and +21 in follicular lymphoma to the changes rea(1)(p36), del(6q), +6, +7, +9, rea(11)(q23), +12, -13 or del(13(q12q14), +18, +21, and +X in DLBCL. The clonal evolution of the secondary aberrations is thought to contribute to the progression of the disease. About 90% (16/18) of other types of NHL had abnormal karyotypes showing specific translocations or gene rearrangements consistent with the pathologic diagnosis. A comprehensive cytogenetics approach including SKY and interphase FISH using probes for specific genes, such as IGH, BCL2, CCND1, and ALK, is a very useful ancillary diagnostic tool for lymphomas. The combined approach also led to the identification of t(2;19)(p23;q13.1) as a new variant of t(2;5)(p23;q35) in a case of Ki-1-positive anaplastic large cell lymphoma with a null cell phenotype. PMID- 12742159 TI - Clonal expansion and not cell interconversion is the basis for the neuroblast and nonneuronal types of the SK-N-SH neuroblastoma cell line. AB - The ability of neuroblastoma (NB) cells to interconvert bidirectionally, in vitro, from a neuroblast (N) to a nonneuronal (S) form is a well-studied biologic phenomenon of great clinical importance. Differences in the morphologic/ biochemical characteristics and gene expression patterns of the two cell populations have been investigated extensively in an effort to unravel the transdifferentiation process. Subcloning of the SK-N-SH NB cell line has led to two morphologically distinct cell types: SH-SY5Y (N-type) and SH-EP (S-type). Karyotypic analysis combined with G-banding and SKY showed a difference between these two cell types in the copy number of the 2p15 approximately pter segment, including the MYC-N gene. FISH analysis showed an extra copy of MYC-N present in all three lines: in SK-N-SH and SH-SY5Y the majority of cells had three copies of MYC-N, whereas in SH-EP the majority had two copies and only a small cell population with three copies was present. We suggest that the simultaneous coexistence of both cell types and the subsequent clonal expansion of one over the other is a possible explanation for the phenomenon observed and not the accepted interconversion model. According to the clonal expansion model, both N and S cells are simultaneously present in both cell lines. Under certain conditions, the less-aggressive S cells can dominate over the highly aggressive N cells, which eventually lead to the formation of the SH-EP and vice-versa. PMID- 12742160 TI - Trisomy 22: a subclone marker? PMID- 12742161 TI - Deletion of 6q23 as sole abnormality in acute myelocytic leukemia. PMID- 12742162 TI - Are cancers dependent on oncogenes or on aneuploidy? PMID- 12742163 TI - ENTH/ANTH domains expand to the Golgi. AB - Clathrin-coated vesicles (CCVs) play important roles in nutrient uptake, downregulation of signaling receptors, pathogen invasion and biogenesis of endosomes and lysosomes. Although detailed models for endocytic CCV formation have emerged, the process of CCV formation at the Golgi and endosomes has been less clear. Key to endocytic CCV formation are proteins containing related phosphoinositide-binding ENTH and ANTH domains. Now, recent studies have identified novel ENTH/ANTH proteins that participate in CCV-mediated traffic between the trans-Golgi Network (TGN) and endosomes and have defined a molecular basis for interaction with AP-1 and GGA adaptors in clathrin coats of the TGN/endosomes. Thus, ENTH/ANTH domain proteins appear to be universal elements in nucleation of clathrin coats. PMID- 12742164 TI - In or out? The dynamics of Smad nucleocytoplasmic shuttling. AB - Members of the TGF-beta family of growth factors mediate their effects through activation of intracellular Smad proteins, which form transcriptionally competent complexes that accumulate in the nucleus. Recently, several reports have demonstrated that Smads actively shuttle between the cytoplasm and nucleus by distinct mechanisms. Here, we summarize what is known about the dynamics of Smad localization and suggest reasons why nucleocytoplasmic shuttling might be important for proper signal transduction. PMID- 12742165 TI - Never say never. The NIMA-related protein kinases in mitotic control. AB - Mitosis sees a massive reorganization of cellular architecture. The microtubule cytoskeleton is reorganized to form a bipolar spindle between duplicated microtubule organizing centers, the chromosomes are condensed, attached to the spindle at their kinetochores, and, through the action of multiple molecular motors, the chromosomes are segregated into two daughter cells. Mitosis also sees a substantial wave of protein phosphorylation, controlling signaling events that coordinate mitotic processes and ensure accurate chromosome segregation. The key switch for the onset of mitosis is the archetypal cyclin-dependent kinase, Cdc2. Under the direction of Cdc2 is an executive of protein serine/threonine kinases that fall into three families: the Polo kinases, Aurora kinases and the NIMA related kinases (Nrk). The latter family has proven the most enigmatic in function, although recent advances from several sources are beginning to reveal a common functional theme. PMID- 12742166 TI - Surfing on microtubule ends. AB - A crowd of proteins seems to have gathered around the plus-ends of microtubules. A rapidly expanding group of proteins known as plus-end tracking proteins (+TIPs) have been identified that seem to be able to 'surf' the dynamic ends of microtubules. Microtubule plus-ends exist in multiple conformational and chemical states. In principle, altering this plus-end microenvironment is an appealing way for regulators such as the +TIPS to control microtubule dynamics; however, specific mechanisms are poorly defined. Here, we focus on new findings addressing the underlying mechanisms of plus-end tracking and the mechanisms by which +TIPS control microtubule dynamics. We review the evidence that plus-end-binding and the control of microtubule dynamics are mechanistically linked. We also consider the possibility that, by studying +TIPs, we might learn more about the dynamic structural changes at the microtubule ends that are at the heart of dynamic instability. PMID- 12742167 TI - Bacterial virulence factors targeting Rho GTPases: parasitism or symbiosis? AB - In the past few years, an important question in microbiology has arisen from reports indicating that several pathogenic bacteria have evolved virulence factors directed towards a Ras subfamily of GTPases, namely the Rho GTPases. Progress made in studying both the virulence factors and the signaling pathways involving Rho GTPases has shed light on this crosstalk. One central question is raised by the findings that both activating and inactivating virulence factors that target Rho GTPases coexist in some pathogenic bacteria. Further studies on this peculiar aspect of the bacteria-host cell interactions, which leads to the outbreak of infectious diseases, might clarify whether this aspect of Rho GTPase activation or inactivation represents a finely adapted response of the pathogen for its own benefit or might lead to a reaction of the host against the bacteria. PMID- 12742168 TI - Constructing an organ: the Drosophila salivary gland as a model for tube formation. AB - Tubes are required in metazoans to transport the liquids and gases that sustain life. The conservation of molecules and mechanisms involved in tube formation suggests that what we learn by studying simple systems will apply to related processes in higher animals. Studies over the past 10 years have revealed the molecules that specify cell fate in Drosophila salivary gland and the cellular events that mediate tube morphogenesis. Here, we discuss how anterior-posterior and dorsal-ventral patterning information specifies both the position of salivary gland primordia and how many cells they contain. We examine the transformation of a polarized epithelial sheet into an elongated, unbranched tube, and the intrinsic and extrinsic factors that influence the final position of the salivary gland. PMID- 12742169 TI - Pre-ribosomes on the road from the nucleolus to the cytoplasm. AB - Eukaryotic ribosomes are assembled in the nucleolus before export to the cytoplasm. Ribosome formation is a highly dynamic and coordinated multistep process, which requires synthesis, processing and modification of pre-rRNAs, assembly with ribosomal proteins and transient interaction of numerous non ribosomal factors with the evolving pre-ribosomal particles. In the past two years, exciting insights into the sequential events occurring during pre-ribosome formation have been obtained, thanks largely to the advances in proteomic analyses. We now have a first biochemical map of the earliest 90S pre-ribosomes and of their daughter pre-40S and pre-60S ribosomal subunits along their path from the nucleolus to the cytoplasm. The future challenge will be to assign functions to the more than 150 non-ribosomal factors that transiently associate with the developing pre-ribosomes. PMID- 12742170 TI - Fibroblast biology in three-dimensional collagen matrices. AB - Research on fibroblast biology in three-dimensional collagen matrices offers new opportunities to understand the reciprocal and adaptive interactions that occur between cells and surrounding matrix in a tissue-like environment. Such interactions are integral to the regulation of connective tissue morphogenesis and dynamics that characterizes tissue homeostasis and wound repair. During fibroblast-collagen matrix remodeling, mechanical signals from the remodeled matrix feed back to modulate cell behavior in an iterative process. As mechanical loading (tension) within the matrix increases, the mechanisms used by cells to remodel the matrix change. Fibroblasts in matrices that are under tension or relaxed respond differently to growth factor stimulation, and switching between mechanically loaded and unloaded conditions influences whether cells acquire proliferative/biosynthetic active or quiescent/resting phenotypes. PMID- 12742171 TI - Chemical genetics: tailoring tools for cell biology. AB - Chemical genetics is a research approach that uses small molecules as probes to study protein functions in cells or whole organisms. Here, I review the parallels between classical genetic and chemical-genetic approaches and discuss the merits of small molecules to dissect dynamic cellular processes. I then consider the pros and cons of different screening approaches and specify strategies aimed at identifying and validating cellular target proteins. Finally, I highlight the impact of chemical genetics on our current understanding of cell biology and its potential for the future. PMID- 12742172 TI - Peak-to-peak dynamics in food chain models. AB - We show in this paper that the chaotic regimes of many food chain models often enjoy a very peculiar property, known as peak-to-peak dynamics. This means that the maximum (peak) density of the populations of any trophic level can be easily forecasted provided the last two peaks of the same population are known. Moreover, extensive simulation shows that only the last peak is needed if the forecast concerns the population at the top of the food chain and that peaks variability often increases from bottom to top. All these findings bring naturally to the conclusion that top populations should be sampled in order to have higher chances to detect peak-to-peak dynamics. The analysis is carried out by studying ditrophic food chain models with seasonally varying parameters, tritrophic food chain models with constant parameters, and more complex food chain and food web models. PMID- 12742173 TI - Optimal foraging and predator-prey dynamics III. AB - In the previous two articles (Theor. Popul. Biol. 49 (1996) 265-290; 55 (1999) 111-126), the population dynamics resulting from a two-prey-one-predator system with adaptive predators was studied. In these articles, predators followed the predictions of optimal foraging theory. Analysis of that system was hindered by the incorporation of the logistic description of prey growth. In particular, because prey self-regulation dependence is a strong stabilizing mechanism, the effects of optimal foraging could not be easily separated from the effects of bottom-up control of prey growth on species coexistence. In this article, we analyze two models. The first model assumes the exponential growth of both prey types while the second model assumes the exponential growth of the preferred prey type and the logistic growth of the alternative prey type. This permits the effect of adaptive foraging on two-prey-predator food webs to be addressed. We show that optimal foraging reduces apparent competition between the two prey types, promotes species coexistence, and leads to multiple attractors. PMID- 12742174 TI - Genealogy-dependent variation in viability among self-incompatibility genotypes. AB - Many hermaphroditic plants avoid self-fertilization by rejecting pollen that express genetically determined specificities in common with the pistil. The S locus, comprising the determinants of pistil and pollen specificity, typically shows extremely high polymorphism, with dozens to hundreds of specificities maintained within species. This article explores a conjecture, motivated by empirical findings, that the expression of recessive deleterious factors at sites closely linked to the S-locus may cause greater declines in the viability of zygotes constituted from more closely related S-alleles. Diffusion approximation models incorporating variation in viability among S-locus genotypes and antagonistic interactions between a new specificity and its immediate parent specificity are constructed and analyzed. Results indicate that variation in viability tends to reduce the number of specificities maintained in a population at stochastic steady state, and that genealogy-based antagonism reduces the rate of bifurcation of S-allele lineages. These effects may account for some of the unusual features observed in empirical studies of S-allele genealogies. PMID- 12742175 TI - Individual-based vs deterministic models for macroparasites: host cycles and extinction. AB - Our understanding of the qualitative dynamics of host-macroparasite systems is mainly based on deterministic models. We study here an individual-based stochastic model that incorporates the same assumptions as the classical deterministic model. Stochastic simulations, using parameter values based on some case studies, preserve many features of the deterministic model, like the average value of the variables and the approximate length of the cycles. An important difference is that, even when deterministic models yield damped oscillations, stochastic simulations yield apparently sustained oscillations. The amplitude of such oscillations may be so large to threaten parasites' persistence.With density dependence in parasite demographic traits, persistence increases somewhat. Allowing instead for infections from an external parasite reservoir, we found that host extinction may easily occur. However, the extinction probability is almost independent of the level of external infection over a wide intermediate parameter region. PMID- 12742176 TI - Steady-state analysis of structured population models. AB - Our systematic formulation of nonlinear population models is based on the notion of the environmental condition. The defining property of the environmental condition is that individuals are independent of one another (and hence equations are linear) when this condition is prescribed (in principle as an arbitrary function of time, but when focussing on steady states we shall restrict to constant functions). The steady-state problem has two components: (i). the environmental condition should be such that the existing populations do neither grow nor decline; (ii). a feedback consistency condition relating the environmental condition to the community/population size and composition should hold. In this paper we develop, justify and analyse basic formalism under the assumption that individuals can be born in only finitely many possible states and that the environmental condition is fully characterized by finitely many numbers. The theory is illustrated by many examples. In addition to various simple toy models introduced for explanation purposes, these include a detailed elaboration of a cannibalism model and a general treatment of how genetic and physiological structure should be combined in a single model. PMID- 12742177 TI - Non-equivalent loci and mutation-selection balance. AB - We consider the implications of mutationally non-equivalent loci for large populations of randomly mating diploid organisms under mutation-selection balance. Variation, across loci, of parameters such as the allelic mutational variance and the mutation rate, is shown to reduce the equilibrium genetic variance. This is proved to follow from the genetic variance contributed by a single locus having an underlying convexity. We give approximate results indicating the way small deviations of the mutational parameters, from their mean values, reduce the genetic variance. Numerical estimates of the size of the effect are given for more general variations of the parameters. Variation in the mutation rates has a significantly smaller effect than variation in the mutational variances. Under accepted parameter values, the reduction in genetic variance can be substantial. PMID- 12742178 TI - Estimating change rates of genetic markers using serial samples: applications to the transposon IS6110 in Mycobacterium tuberculosis. AB - In infectious disease epidemiology, it is useful to know how quickly genetic markers of pathogenic agents evolve while inside hosts. We propose a modular framework with which these genotype change rates can be estimated. The estimation scheme requires a model of the underlying process of genetic change, a detection scheme that filters this process into observable quantities, and a monitoring scheme that describes the timing of observations. We study a linear "birth-shift death" model for change in transposable element genotypes, obtaining maximum likelihood estimators for various detection and monitoring schemes. The method is applied to serial genotypes of the transposon IS6110 in Mycobacterium tuberculosis. The estimated birth rate of 0.0161 (events per copy of the transposon per year) and death rate of 0.0108 are both significantly larger than the estimated shift rate of 0.0018. The sum of these estimates, which corresponds to a "half-life" of 2.4 years for a typical strain that has 10 copies of the element, substantially exceeds a previous estimate of 0.0135 total changes per copy per year. We consider experimental design issues that enable the precision of estimates to be improved. We also discuss extensions to other markers and implications for molecular epidemiology. PMID- 12742180 TI - Adolescent endometriosis: diagnosis and treatment approaches. AB - OBJECTIVES: To review the etiologies, diagnosis, and treatment options of adolescent endometriosis. METHODS: Review of publications relating to adolescent endometriosis. RESULTS: Endometriosis occurs in adolescents as young as 8 years of age; furthermore, there have been documented cases of endometriosis occurring prior to menarche. Adolescents presenting with pelvic pain are treated with cyclic combination oral contraceptive pills and nonsteroidal anti-inflammatory agents. If the pain does not respond to these therapies, then in adolescents as in adults, an operative laparoscopy is recommended for the diagnosis and surgical management of endometriosis. The operating gynecologist should be familiar with the appearance of the complete spectrum of various morphologies of endometriosis, as adolescents tend to have clear, red, white, and/or yellow-brown lesions more frequently than black or blue lesions. Subtle clear lesions of endometriosis may be better visualized by filling the pelvis with irrigation fluid so that the clear lesions can be appreciated in a three-dimensional appearance. Young women who are found to have endometriosis by laparoscopy may present with acyclic, cyclic, and constant pelvic pain. Adolescents with pelvic pain not responding to conventional medical therapy have approximately a 70% prevalence of endometriosis. It is known that endometriosis is a progressive disease and since there is no cure, adolescents with endometriosis require long-term medical management until the time in their lives when they have completed childbearing. Psychosocial support is extremely important for this population of young women with endometriosis. CONCLUSIONS: Endometriosis occurs in adolescents, and presenting symptoms may vary from those seen in adult women with the disease. All health care providers must be aware of the existence of adolescent endometriosis. They should also be aware of the presenting symptoms so that the adolescent can be appropriately referred to a gynecologist comfortable with medical and surgical treatment options in this patient population. If laparoscopy is to be undertaken, the gynecologist must be prepared not only to diagnose but to surgically manage endometriosis. In addition, the subtle laparoscopic findings of endometriosis in adolescents must be recognized for an appropriate diagnosis. Long-term medical therapy will hopefully decrease pain and the progression of the disease, thus decreasing the risk of advanced-stage disease and infertility. PMID- 12742181 TI - Psychosocial aspects of teen endo. PMID- 12742182 TI - Management of adolescent chronic pelvic pain from endometriosis: a pain center perspective. AB - OBJECTIVES: To review the options for the treatment of chronic pelvic pain in adolescents with endometriosis. METHODS: Review of publications and description of author's experience relating to adolescent endometriosis and pain treatment. RESULTS: Pain treatment services offer a multidisciplinary approach for the treatment of chronic pelvic pain consisting of analgesic trials, cognitive behavioral therapy, physical therapy, and complementary and alternative therapies. Patients are also evaluated for other conditions that might contribute to their chronic pain. Medication trials of antidepressants, anticonvulsants, tramadol, and nonsteroidal anti-inflammatory drugs are frequently used; however, there are limited data on efficacy. Biobehavioral techniques such as relaxation and biofeedback can help patients modify their experience of pain. Cognitive behavioral therapy can improve coping skills and promote improved functioning, particularly in patients who are debilitated from chronic pain. Transcutaneous electrical stimulation (TENS) and physical therapy approaches may be helpful in patients with musculoskeletal conditions that contribute to their pelvic pain. Complementary and alternative therapies are frequently used in the treatment of chronic pain; controlled trials are needed to assess efficacy. CONCLUSIONS: An individualized, multidisciplinary approach may be effective in improving overall outcome in patients with chronic pelvic pain in reducing pain and normalizing function. PMID- 12742183 TI - Big picture of endometriosis helps provide guidance on approach to teens: comparative historical data show endo starting younger, is more severe. PMID- 12742184 TI - Tips on treating teens with endometriosis. PMID- 12742185 TI - MK-801 inhibits the micturition reflex in chronic bladder irritation caused by crystalluria in the rat. AB - Urodynamic and pharmacological studies were performed to investigate the effect of crystalluria on the micturition reflex and the involvement of glutamatergic transmission. The rats, which were given LP-805 (100 mg/kg/day) orally for 12 days, voided crystalluria. The pH of these crystalluria (LP-805 urine) was the same as normal urine. The amount of crystals was 70-100/division magnified 400 x. The end of the crystals was sharp. Intravesical administration of LP-805 urine induced hyperreflexia of the micturition reflex in normal rats. When the infusion solution was changed to LP-805 urine from saline, the latency was reduced to 57.6+/-2.1% of control in single cystometrogram (CMG) or was reduced to 51.4+/ 0.9% of control in continuous CMG. The voiding volume was reduced to 52.1+/-3.6% of control in single CMG or was reduced to 62.5+/-0.8% of control in continuous CMG. These parameters were recovered after LP-805 urine was removed. Intravesical administration of acetic acid did not induce hyperreflexia of the micturition reflex in LP-805-treated rats. These data suggest that the chronic irritation by aculeate crystals might induce hyperreflexia of the micturition reflex, which increase afferent neuronal activity. Intravenous administration of MK-801 (0.001 to 1 mg/kg) inhibited the micturition reflex in a dose-dependent manner. The ID50 in LP-805-treated rats (0.03 mg/kg i.v.) was lower than that in normal rats (0.56 mg/kg i.v.). After chronic irritation of the bladder epithelium, MK-801 sensitivity was enhanced for the micturition reflex. These data suggested that crystalluria elicit hyperreflexia in the micturition reflex that mediated with NMDA glutamatergic receptors. PMID- 12742186 TI - Tyrosine hydroxylase and norepinephrine transporter in sympathetic ganglia of female rats vary with reproductive state. AB - In females, sympathetic activity varies with changes in reproductive status, but whether expression of proteins critical to the function of sympathetic neurons is also altered is unknown. Therefore, the present study tested the hypothesis that, in rat adrenal gland and superior cervical ganglia, the expression of tyrosine hydroxylase (TH) and the norepinephrine transporter (NET), measured using Western analysis, are changed during pregnancy and the estrous cycle. Compared to diestrus, pregnancy increased TH levels in both superior cervical ganglia and adrenal gland. Pregnancy was also associated with decreased NET levels in the superior cervical ganglia, but increased levels in the adrenal gland. Relative to diestrus, the pattern of changes of TH and the NET in rats during proestrus was generally similar to changes observed during pregnancy. To assess whether gonadal hormones were involved, ovariectomized rats were also studied and changes in serum estrogen and progesterone were assayed in a subset of animals in all groups. Variations in TH and the NET among all groups did not correlate with changes in either estrogen or progesterone, suggesting that the steroids were not exclusively responsible. In conclusion, reproductive status alters the expression of TH and the NET in adrenal gland and superior cervical ganglia of female rats, which could significantly influence the function of the sympathetic nervous system. However, the mechanism for these changes does not depend solely on changes in estrogen or progesterone. PMID- 12742187 TI - Cholinergic pathways modulate experimental dinitrobenzene sulfonic acid colitis in rats. AB - Recent studies have suggested that neuroimmune interactions modulate intestinal mucosal immune responses. In the current study, we examined the role of cholinergic pathways in modulating the severity of acute dinitrobenzene sulfonic acid colitis, using pharmacological agents to suppress acetylcholinesterase in Sprague-Dawley rats, and evaluating the colitis in the cholinergic hyperresponsive Flinder's sensitive line rats and their control counterparts, the Flinder's resistant line. Colitis was induced by intrarectal dinitrobenzene sulfonic acid (80 mg x ml(-1) in 50% ethanol); controls received intrarectal saline. Sprague-Dawley rats received an acetylcholinesterase inhibitor, physostigmine (50 microg x kg(-1) s.c.) or neostigmine (50 microg x kg(-1) s.c.), 30 min prior to intrarectal dinitrobenzene sulfonic acid; controls received saline vehicle. On day 5, the macroscopic damage score, myeloperoxidase activity (an estimate of granulocyte infiltration) and smooth muscle thickness were evaluated in the inflamed colonic segment. Significant increases in macroscopic damage score and colonic smooth muscle thickness were observed in Sprague-Dawley and Flinder's Resistant Line rats on day 5 following intrarectal dinitrobenzene sulfonic acid compared to saline controls. Increased myeloperoxidase activity was also observed in dinitrobenzene sulfonic acid-treated Sprague-Dawley rats and Flinder's Resistant Line rats. In contrast, Flinder's Sensitive Line rats failed to demonstrate a significant rise in macroscopic damage, smooth muscle layer thickness, or myeloperoxidase activity on day 5 following intrarectal dinitrobenzene sulfonic acid when compared to saline-treated Flinder's Sensitive Line controls. Neostigmine and physostigmine treatment prior to intrarectal dinitrobenzene sulfonic acid significantly attenuated macroscopic damage score, myeloperoxidase activity and smooth muscle thickness on day 5 compared to colitic Sprague-Dawley controls. Significantly greater reductions in myeloperoxidase activity were observed with physostigmine vs. neostigmine pretreatment. These data suggest that cholinergic pathways modulate the acute colonic inflammatory response associated with the dinitrobenzene sulfonic acid model, with central pathways exerting a greater protective effect relative to peripheral pathways. Further studies are required to determine the contributions of sites in the nervous system and neuro-effector junctions. PMID- 12742188 TI - Baroreceptor-sensitive neurons in the rat paratrigeminal nucleus. AB - The paratrigeminal nucleus (Pa5) is a small collection of medullary neurons localized in the dorsal lateral spinal trigeminal tract. Electrophysiological and anatomical studies showed functional Pa5 efferent connections to the rostroventrolateral reticular nucleus (RVL) and the nucleus of the solitary tract (NTS), both well-studied components of the baroreflex arch. Similarly to the NTS, the main site for termination of cardiovascular peripheral afferents, the Pa5 receives primary sensory inputs of glossopharyngeal and vagus nerves, which suggests that the Pa5 may play a role in the baroreceptor reflex modulation. Simultaneous recording from multiple single neurons in 10 freely behaving rats showed that 37% of recorded Pa5 neurons altered firing rates (35% increased and 2% decreased) during the peak arterial blood pressure response to i.v. phenylephrine. Forty two percent of the 84 identified Pa5 baroreceptor-excited neurons showed high correlation to cardiac cycle denoting the synchronous phasicity to fast changes of blood pressure. Autocorrelation analysis revealed that 48 pressure-sensitive and 55 nonpressure-sensitive neurons have periodical activities which were not directly linked to cardiac cycle. We suggest that the Pa5, a yet unknown component of the baroreflex pathway, may relay baroreceptor information to the NTS and by passing other components of the baroreceptor reflex arch, directly to sympathetic premotor neurons in the RVL. PMID- 12742190 TI - Differential sensitivity to calcium and osmotic pressure of fast and slow ATP currents at sympathetic varicosities in mouse vas deferens. AB - Secretion of noradrenaline from large dense-core vesicles in chromaffin cells involves both rapid and slow components of exocytosis which are differentially sensitive to changes in external calcium, osmotic pressure and interruption of the interacting SNARE proteins. Electrical signs of secretion of ATP from sympathetic nerve terminals of mouse vas deferens, the excitatory junctional currents (EJCs), also indicate both rapid and slow mechanisms of exocytosis, which might also show such differential sensitivity. We report here that the large and fast EJCs are highly sensitive to changes in extracellular calcium ions whereas the small and slow EJCs are not. Furthermore, the frequency of fast EJCs is accelerated by hypotonic solutions whereas the slow EJCs are accelerated by hypertonic solution. Fast EJCs, but not slow EJCs, are blocked by peptide fragments of alpha-SNAP and syntaxin whereas slow EJCs are not. These observations point to two classes of exocytosis from sympathetic nerve terminals that parallel those of exocytosis from chromaffin cells. PMID- 12742189 TI - Long-term potentiation of nicotinic synaptic transmission in rat superior cervical ganglia produced by phorbol ester and tetanic stimulation. AB - The long-term potentiation of nicotinic synaptic transmission induced by both active phorbol ester (4beta-phorbol-12,13-dibutyrate, PdBu) and tetanic trains of preganglionic stimulation was studied in single neurons of the superior cervical ganglion (SCG) of the rat using intracellular recording techniques. PdBu significantly increased the mean amplitude of both the unitary evoked fast excitatory postsynaptic potentials (EPSPs) and the fast excitatory postsynaptic currents (EPSCs) to 17.0+/-3.3 mV (control 8.4+/-1.9 mV, n=5) and 2.8+/-0.4 nA (control 0.8+/-0.1 nA, n=10), respectively. There was no significant change in either the resting membrane potential, input resistance, or the threshold for the initiation of an action potential. The response to exogenously applied acetylcholine (ACh) was also not changed following exposure to PdBu. In low calcium, high-magnesium solutions, PdBu significantly increased the quantal content of EPSPs approximately threefold from a control of 0.9+/-0.2 (n=5) to 2.6+/-0.6 (n=5). The quantal content of EPSCs was also increased to 1.3+/-0.2 (control 0.5+/-0.1, n=10). PdBu increased the frequency of miniature EPSPs (mEPSPs) to 196+/-47% (n=6) of control, while the amplitude, rise time, rate of rise, and decay of mEPSPs were not significantly changed. Tetanic stimulation significantly increased the amplitude of the unitary synaptic EPSPs and EPSCs without significantly changing the resting membrane potential, input resistance, threshold for initiation of an action potential, or the response to exogenously applied ACh. Tetanic stimulation significantly increased quantal content of EPSPs and EPSCs threefold. The results obtained with tetanically induced LTP are similar to the results obtained with phorbol ester-induced LTP in these ganglion neurons. These results suggest that both tetanically induced and phorbol ester induced LTP, in the rat, share similar mechanisms which involve, at least in part, activation of PKC-dependent mechanisms to increase quantal release from sympathetic preganglionic axon terminals. PMID- 12742191 TI - Can extremely low frequency alternating magnetic fields modulate heart rate or its variability in humans? AB - This study is a reexamination of the possibility that exposure to extremely low frequency alternating magnetic field (ELF-MF) may influence heart rate (HR) or its variability (HRV) in humans. In a wooden room (cube with 2.7-m sides) surrounded with wire, three series of experiments were performed on 50 healthy volunteers, who were exposed to MFs at frequencies ranging from 50 to 1000 Hz and with flux densities ranging from 20 to 100 microT for periods ranging from 2 min to 12 h. In each experiment, six indices of HR/HRV were calculated from the RR intervals (RRIs): average RRI, standard deviation of RRIs, power spectral components in three frequency ranges (pVLF, pLF and pHF), and the ratio of pLF to pHF. Statistical analyses of results revealed no significant effect of ELF-MFs in any of the experiments, and suggested that the ELF-MF to which humans are exposed in their daily lives has no acute influence on the activity of the cardiovascular autonomic nervous system (ANS) that modulates the heart rate. PMID- 12742192 TI - Occurrence of the spinal reflex due to skin pressure in sudomotor and cutaneous vasoconstrictor nerve system of humans. AB - The effects of skin pressure applied to one side of the waist on sudomotor and vasoconstrictor nerve activity were compared with the effects on sweating and cutaneous blood flow in humans. The sweat rate and cutaneous blood flow were measured on left and right dorsal feet. Skin sympathetic nerve activity (SSNA) was recorded by microneurography from a microelectrode inserted in left and right peroneal nerves. Skin pressure was applied in a supine position to the area over the left or right anterior superior iliac spine under warm (T(a): 30-36 degrees C) and cool (T(a): 19-23 degrees C) conditions. Sudomotor and vasoconstrictor bursts were identified for quantitative analysis. The skin pressure increased the contralateral/ipsilateral ratio of the sweat rate. It also increased the contralateral/ipsilateral ratio of the cutaneous blood flow and the contralateral/ipsilateral ratio of the sudomotor burst amplitude. However, skin pressure did not induce any significant changes in the contralateral/ipsilateral ratio of the vasoconstrictor burst amplitude. The results indicate that an asymmetrical reflex effect of skin pressure on vasoconstrictor nerve activity was absent, suggesting that, whereas the ipsilateral suppression of sweating elicited by skin pressure was mediated by the sudomotor nerve system, the ipsilateral suppression of cutaneous blood flow was not mediated by the vasoconstrictor nerve system. Thus, the occurrence of the spinal reflex due to skin pressure is not uniform between the sudomotor and the vasoconstrictor nerve systems, which represent different organizations at the level of spinal cord. PMID- 12742193 TI - Attenuation of cerebral vasospasm in rabbits using clonidine hydrochloride, a central adrenergic agonist. AB - The aim of this study was to assess, firstly, if exclusion of central noradrenergic areas in the hypothalamus and brain stem with the central sympathetic blocker clonidine hydrochloride could prevent the development of chronic vasospasm following experimental subarachnoid haemorrhage in rabbits and, secondly, if, parallel with the effect on cerebral arteries, changes in dopamine beta-hydroxylase concentration in the hypothalamus and brain stem could also be detected. Experimental subarachnoid haemorrhage, in concentrations of 1 ml of autologous arterial blood/1 kg of body weight was carried out on 18 New Zealand rabbits. Histological specimens were obtained by the method of perfusion fixation after the rabbits were sacrificed on day 8 after subarachnoid haemorrhage. The spastic effect of experimentally induced subarachnoid haemorrhage was determined by assessing the intensity of corrugation of the intima of the rabbit basilar artery by the previously developed method of corrugation coefficient and computer image analysis. The concentration and localization of dopamine beta-hydroxylase in noradrenaline-containing neurons was immunohistochemically assessed (semiquantitatively as 0, 1 and 2) with anti-dopamine beta-hydroxylase, at precisely defined sites of the hypothalamus and brain stem of the same rabbit. The results revealed less corrugated and smoother intima in the basilar artery and significantly lower dopamine beta-hydroxylase concentration in the control group of rabbits with sham subarachnoid haemorrhage and without any additional interventions (mean corrugation coefficient=1.123+/-0.024, P=0.35 x 10(-3); mean dopamine beta-hydroxylase=0.350+/-0.071, P=0.22 x 10(-3)), and smoother intima in the basilar artery with significantly lower concentration of dopamine beta hydroxylase in the clonidine group (rabbits with subarachnoid haemorrhage and central alpha(2)-blocker clonidine hydrochloride at a daily dose of 0.03 mg/kg of body weight for 8 days; mean corrugation coefficient=1.177+/-0.058, P=1.7 x 10( 3); mean dopamine beta-hydroxylase=0.583+/-0.175, P=1.1 x 10(-3)). In comparison, the haemorrhage only group (rabbits with subarachnoid haemorrhage and without medication; mean corrugation coefficient=1.370+/-0.101; mean dopamine beta hydroxylase=1.214+/-0.313) displayed intensive corrugation of the intima of the basilar artery and a significantly more intensive accumulation of dopamine beta hydroxylase than did the control group and the clonidine group. The results of this study demonstrated that the central alpha(2)-blocker clonidine hydrochloride effectively prevents vasospasm, and diminishes the concentration of cerebral dopamine beta-hydroxylase in the hypothalamus and brain stem after experimental subarachnoid haemorrhage in rabbits. PMID- 12742194 TI - Complications and costs after high-risk surgery: where should we focus quality improvement initiatives? AB - BACKGROUND: Data on the relative clinical and economic impact of postoperative complications are needed in order to direct quality improvement efforts. STUDY DESIGN: Patients undergoing two high-risk surgical procedures, hepatectomy (n = 569) and esophagectomy (n = 366), from 1994 to 1998 were included. Data were abstracted from the Maryland hospital discharge database. Relative resource use was determined using median regression, adjusting for patient comorbidities and other case-mix variables. RESULTS: A total of 935 patients were studied. Overall in-hospital mortality was 6.1%; complication rate was 38.4%. Median cost for all patients was $14,527 (interquartile range $10,936-$21,412) and length of stay 9 days (interquartile range 7-13 days). Median hospital cost was increased for patients with complications ($16,868 versus $12,861; p < 0.001). In the multivariate analysis, several complications remained associated with increased cost. Acute renal failure ($25,219), septicemia ($18,852), and myocardial infarction ($9,573) were associated with the greatest increase in resource use. But because the incidence of each complication varies, the attributable fraction of total resource use was highest for acute renal failure (19%), septicemia (16%), and surgical complications (16%). CONCLUSIONS: Complications are independently associated with increased resource use after high-risk surgery. Population-based studies may be valuable in determining the relative economic importance of postoperative complications. Quality improvement efforts for these complications should be prioritized based on both the incidence of the complication and its independent contribution to increased resource use. PMID- 12742195 TI - Does tachycardia correlate with hypotension after trauma? AB - BACKGROUND: Tachycardia is believed to be closely associated with hypotension and is often listed as an important sign in the initial diagnosis of hemorrhagic shock, but the correlation between heart rate and hypotension remains unproved. STUDY DESIGN: Data were collected from all trauma patients, 16 to 49 years old, presenting to our university-based trauma center between July 1988 and January 1997. Moribund patients with a systolic blood pressure < or =50 or heart rate < or = 40 and patients with significant head or spinal cord injuries were excluded. Tachycardia was defined as a heart rate >or= 90 and hypotension as a systolic blood pressure < 90. RESULTS: Hypotension was present in 489 of the 14,325 admitted patients that met the entry criteria. Of the hypotensive patients, 35% (169) were not tachycardic. Tachycardia was present in 39% of patients with systolic blood pressure 120 mmHg. Hypotensive patients with tachycardia had a higher mortality (15%) compared with hypotensive patients who were not tachycardic (2%, P = 0.003). Logistic regression analysis revealed tachycardia to be independently associated with hypotension (p = 0.0004), but receiver operating curve analysis demonstrated that the sensitivity and specificity of heart rate for predicting hypotension is poor. CONCLUSIONS: Tachycardia is not a reliable sign of hypotension after trauma. Although tachycardia was independently associated with hypotension, its sensitivity and specificity limit its usefulness in the initial evaluation of trauma victims. Absence of tachycardia should not reassure the clinician about the absence of significant blood loss after trauma. Patients who are both hypotensive and tachycardic have an associated increased mortality and warrant careful evaluation. PMID- 12742196 TI - Organ-specific changes in high-energy phosphates after hemorrhagic shock and resuscitation in the rat. AB - BACKGROUND: Adenosine nucleotides provide energy for many essential cellular functions. Liver and intestinal ATP and energy charge are known to decrease during hemorrhagic shock, and the ability to regenerate high-energy phosphates may have important implications for recovery. We measured organ-specific changes in energy charge after hemorrhagic shock and after shock followed by resuscitation. STUDY DESIGN: Anesthetized Sprague-Dawley rats were bled and maintained at a mean arterial pressure (MAP) of 40 mmHg for 1, 2, 3, or 4 hours. Some animals were resuscitated with normal saline and shed blood (1:1) to a mean arterial pressure of 80 to 90 mmHg for 1 hour. Control animals were anesthetized, but not hemorrhaged. At the conclusion, blood gases and adenine nucleotides were measured. RESULTS: Arterial pO2 and pCO2 were normal in all groups. Unresuscitated hemorrhage caused metabolic acidosis, but bicarbonate was normal in controls and after hemorrhage followed by resuscitation. Energy charge (EC) in the gastrocnemius was unaffected by hemorrhage or resuscitation. Liver EC decreased after hemorrhage (p = 0.0001), but recovered partially after resuscitation. Kidney EC was decreased after only 3 hours of hemorrhage and 1 hour of resuscitation (p = 0.005), but not with shorter periods of hemorrhage. Lung EC decreased with shock, but was substantially worse after resuscitation (p < 0.05). CONCLUSIONS: After hemorrhage and resuscitation, EC decreased in lung, liver, kidney, and intestine, but the time course, extent of decline, and ability to recover after resuscitation varied from organ to organ. Inability to regenerate high-energy phosphates after hemorrhagic shock may be a marker for more severe cellular damage. PMID- 12742197 TI - The effect of recombinant factor VIIa on noncoagulopathic pigs with grade V liver injuries. AB - BACKGROUND: Recombinant Factor VIIa (rFVIIa) has been used to decrease bleeding in a number of settings, including hemophilia, liver transplantation, intractable bleeding, and cirrhosis. It has also been shown to reduce bleeding in coagulopathic pigs with Grade V liver injuries when used as an adjunct to packing. This study was performed to determine if rFVIIa would reduce blood loss after a Grade V liver injury in noncoagulopathic pigs when used as sole therapy. STUDY DESIGN: Thirty normothermic animals were randomized to receive either 150 microg/kg of rFVIIa or normal saline intravenously. After laparotomy and splenectomy, a standardized Grade V liver injury was made with a liver clamp. Thirty seconds after injury, blinded therapy was given. Blood loss was measured 15 minutes after injury and the abdomen was closed. Animals were resuscitated to their baseline blood pressure and the study was continued for 2 hours. Serial coagulation parameters were obtained. Following the study period, blood loss was measured and an autopsy was performed. Grossly normal areas of lung were examined for evidence of intravascular thrombosis. RESULTS: Mean Factor VII:C levels increased 155-fold in the treatment group after infusion of rFVIIa. The mean prothrombin time in the treatment group decreased from 9.8 +/- 0.4 seconds to 7.3 +/- 0.2 seconds and remained significantly different from the control group throughout the study (p < 0.01). There were no differences in other coagulation parameters. Mean initial blood loss was 822 +/- 266 mL in the treatment group and 768 +/- 215 mL in the control group (p = 0.6). Rebleeding blood volume was 397 +/ 191 mL in the treatment group and 437 +/- 274 mL (p = 0.6) in the control group. Lung histology revealed no evidence of abnormal microvascular thrombosis. CONCLUSIONS: rFVIIa does not reduce blood loss after Grade V liver injury when it is used as sole therapy in warm noncoagulopathic pigs. PMID- 12742198 TI - Impact of minimally invasive surgery on the treatment of esophageal achalasia: a decade of change. AB - BACKGROUND: Twenty years ago an average of 1.5 Heller myotomies were performed per year in our hospital, mostly for patients whose dysphagia did not improve following balloon dilatation or whose esophagus had been perforated during a balloon dilatation. Ten years ago we started using minimally invasive surgery to treat this disease. STUDY DESIGN: This study measures the impact of minimally invasive surgery with regard to the following: the number of patients referred for treatment; the number of patients who came to surgery without previous treatment; and the results of surgical treatment. Between 1991 and 2001, 149 patients had minimally invasive surgery for achalasia: 25 patients (17%) had thoracoscopic Heller myotomy and 124 (84%) had laparoscopic Heller myotomy and Dor fundoplication. Of the 149 patients, 79 patients (53%) had previous treatment (56 patients [71%], balloon dilatation; 7 patients [9%], botulinum toxin injection; 16 patients [20%], both) and 70 patients (43%) had none of these treatments. Mean postoperative followup was 59 +/- 36 months. Patients were divided into two groups: group A, operated on between 1991 and 1995; and group B, operated on between 1996 and 2001. RESULTS: In the past decade, the number of patients referred for surgery has increased substantially--group A, 48; group B, 101; an increasing proportion of patients were referred for surgery without previous treatment--group A, 38%; group B, 51%; and the outcomes of the operation progressively improved--group A, 87%; group B, 95%. CONCLUSIONS: These data show that the high success rate of laparoscopic Heller myotomy for achalasia has brought a shift in practice; surgery has become the preferred treatment of most gastroenterologists and other referring physicians. This has followed documentation that laparoscopic treatment outperforms balloon dilatation and botulinum toxin injection. PMID- 12742201 TI - Barrett's esophagus can and does regress after antireflux surgery: a study of prevalence and predictive features. AB - BACKGROUND: To investigate the factors leading to histologic regression of metaplastic and dysplastic Barrett's esophagus (BE). STUDY DESIGN: The study sample consisted of 91 consecutive patients with symptomatic Barrett's esophagus. Pre- and posttreatment endoscopic biopsies from 77 Barrett's patients treated surgically and 14 treated with proton pump inhibitors (PPI) were reviewed. An expert pathologist confirmed the presence of intestinal metaplasia (IM) with or without dysplasia. Posttreatment histology was classified as having regressed if two consecutive biopsies taken more than 6 months apart plus all subsequent biopsies showed loss of IM or loss of dysplasia. Clinical factors associated with regression were studied by multivariate analysis, as was the time course of its occurrence. RESULTS: Histopathologic regression occurred in 28 of 77 patients (36.4%) after antireflux surgery and in 1 of 14 patients (7.1%) treated with PPIs alone (p < 0.03). After surgery, regression from low-grade dysplastic to nondysplastic BE occurred in 17 of 25 patients (68%) and from IM to no IM in 11 of 52 (21.2%). Both types of regression were significantly more common in short (< 3 cm) than long (> 3 cm) segment Barrett's esophagus; 19 of 33 (58%) and 9 of 44 (20%) patients, respectively (p = 0.0016). Eight patients progressed, five from IM alone to low-grade dysplasia and three from low- to high-grade dysplasia. All those who progressed had long segment BE. On multivariate analysis, presence of short segment Barrett's and type of treatment were significantly associated with regression; age, gender, surgical procedure, and preoperative lower esophageal sphincter and pH characteristics were not. The median time of biopsy proved regression was 18.5 months after surgery, with 95% occurring within 5 years. CONCLUSIONS: This study refutes the widely held assumption that once established, Barrett's esophagus does not change. More than one-third of patients with visible segments of Barrett's esophagus undergo histologic regression after antireflux surgery. Regression is dependent on the length of the columnar-lined esophagus and time of followup after antireflux surgery. PMID- 12742203 TI - Quality of life improves within 30 days of surgery for Crohn's disease. AB - BACKGROUND: The effect of surgery on quality of life (QOL) in the early postoperative period is important in Crohn's disease because of the multiple surgical procedures that patients undergo and the acute QOL benefits that might occur as a result of modifications of medical treatment. Earlier studies of the effect of surgery on QOL have been retrospective and assessed changes 3 to 24 months after surgery. This study prospectively assesses the effect of surgery on QOL in the early postoperative period. STUDY DESIGN: Patients requiring surgical management of sequelae of Crohn's disease were obtained from a prospectively entered database including data on QOL. Preoperative and 30-day postoperative QOL were determined in 82 patients using Cleveland Global Quality of Life (CGQL) scores (range from 0 [worst] to 10 [best possible] QOL). Preoperative and postoperative scores were compared using a paired t-test to determine the significance of any change in QOL after surgery. The effect of other variables on change in QOL after surgery was assessed using the t-test or analysis of variance. Multifactor analysis of variance was used to assess the effect of several independent variables. RESULTS: Eighty-two patients (41 women) of 142 patients who had had surgery (58%) had complete preoperative and 30-day postoperative scores. The incidence of complications was 23% (11% were major). There was a significant improvement in QOL 30 days after surgery as measured by CGQL (0.6 +/- 0.2 preoperative to 0.7 +/- 0.2 postoperative; mean +/- SD; p < 0.001). The mean preoperative CGQL was 0.56 +/- 0.24 and the mean improvement was 0.11 +/- 0.20 toward a better QOL. Female patients (p < 0.05) and those who did not develop complications within 30 days of surgery (p < 0.05) had a significantly greater improvement in CGQL after surgery than other groups. No other factor was predictive of improved outcomes. CONCLUSIONS: QOL as measured by CGQL improves early after surgery (30 days postoperatively). Improvement in CGQL is greater in female patients and patients who do not develop complications in the postoperative period. It is not affected by other patient characteristics, nature of disease, indication, or procedure performed. Most patients who undergo surgery for Crohn's disease feel that surgery has helped them and would undergo surgery again. PMID- 12742204 TI - Elective bowel resection for incurable stage IV colorectal cancer: prognostic variables for asymptomatic patients. AB - BACKGROUND: Surgical resection of primary colorectal cancer (CRC) in patients with stage IV disease at initial presentation remains controversial. Although bowel resection to manage symptoms such as bleeding, perforation, or obstruction has been advocated, management of asymptomatic patients has not been well defined. Patient-dependent factors (performance status, comorbid disease) and extent of distant metastases are among the considerations that impact on the decision to proceed with surgical management in asymptomatic stage IV CRC patients. We postulated that selected patients might benefit from elective resection of the asymptomatic primary CRC. The extent of distant metastases was objectively measured by several methods to identify potential prognostic variables that may help guide patient selection in this population. STUDY DESIGN: We reviewed hospital and colorectal service databases for the years 1996 to 1999. Stage IV patients who had colorectal resections with gross residual metastatic disease were identified (n = 209). Among these 209 patients, 82 patients operated on for symptoms (obstruction, perforation, bleeding, or pain) were excluded, leaving 127 patients who underwent elective resection of their asymptomatic primary CRC. Over the same time period, 103 stage IV patients who did not undergo resection were identified. Data on patient characteristics and clinical management were collected. A radiologist performed an independent review of available CT scans to assess extent of liver disease. The chi-square test was used for analysis of categoric data and Student's t-test for continuous variables. Survival was determined by the Kaplan-Meier method and distributions compared by the log rank test. Multivariate analysis was performed using Cox regression. RESULTS: The resected group could be easily distinguished from the nonresected group by a higher frequency of right colon cancers (p = 0.03) and metastatic disease restricted to the liver (p = 0.02) or one other site apart from the primary tumor (p = 0.02). Resected patients had prolonged median (16 versus 9 months, p < 0.001) and 2-year (25% versus 6%, p < 0.001) survival compared with patients never resected. Univariate analysis identified three significant prognostic variables (number of distant sites involved, metastases to liver only, and volume of hepatic replacement by tumor) in the resected group. Volume of hepatic replacement was also a significant predictor of survival in Cox multivariate regression analysis (p = 0.01). Subsequent to resection of asymptomatic primary CRC, 26 patients (20%) developed postoperative complications. Median hospital stay was 6 days. Two patients (1.6%) died within 30 days of surgery. CONCLUSION: Stage IV patients selected for elective palliative resection of asymptomatic primary colorectal cancers had substantial postoperative survival that was significantly better than those never having resection. Limited metastatic tumor burden and less extensive liver involvement were associated with better survival and a higher likelihood of benefit from elective bowel resection in asymptomatic patients with incurable stage IV CRC. PMID- 12742205 TI - Study of the role of the second defecation reflex: anorectal excitatory reflex in the pathogenesis of constipation. AB - BACKGROUND: Previous studies have shown that anal distension caused rectal contraction, an action mediated through the anorectal excitatory reflex. Anal anesthetization aborted rectal contraction and rectal evacuation was induced by excessive straining. We investigated the hypothesis that inhibition or absence of the anorectal excitatory reflex could lead to constipation. METHODS: We studied 18 patients (mean age +/- SD: 40.6 +/- 5.8 years, 14 women) with rectal inertia, 14 (41.7 +/- 6.6 years, 12 women) with puborectalis paradoxical syndrome, and 10 healthy volunteers (37.9 +/- 4.8 years, 8 women). The rectum was filled with normal saline until urge and then evacuated; residual fluid was calculated. The anal and rectal pressure response to anal balloon distension in increments of 2 mL of saline was recorded by a two-channel microtip catheter. RESULTS: In the healthy volunteers, saline was evacuated as a continuous stream without straining except occasionally at the start of evacuation; no residual fluid was encountered. Anal balloon distension effected notable rectal pressure increase. In rectal inertia patients, evacuation occurred in small fluid gushes produced with excessive straining; residual fluid of large volume was collected. Anal balloon distension up to 10 mL produced no notable rectal pressure changes. The patients with PPS failed to evacuate more than a few mL of fluid despite excessive straining; the volume of residual fluid was considerable. Anal balloon distension caused a notable rectal pressure rise. The results were reproducible. CONCLUSIONS: These results suggest that the defecation reflexes (rectoanal and anorectal) are absent in rectal inertia patients and this presumably denotes a neurogenic disorder. The anorectal reflex is active in puborectalis paradoxical syndrome, but the rectoanal reflex is not, indicating a possible myogenic defect in the puborectalis muscle. PMID- 12742206 TI - Introduction: inguinal hernia management-testing management strategies in two clinical trials. PMID- 12742207 TI - The development of a clinical trial to determine if watchful waiting is an acceptable alternative to routine herniorrhaphy for patients with minimal or no hernia symptoms. AB - BACKGROUND: This article describes the development and implementation of a randomized clinical trial designed to answer the question: Is watchful waiting an acceptable alternative to operation for men with asymptomatic or minimally symptomatic inguinal hernias? STUDY DESIGN: A clinical trial has been designed to compare watchful waiting and operation for men with an asymptomatic or a relatively asymptomatic inguinal hernia. Men are randomized to watchful waiting or a standard open operation, the Lichtenstein tension-free hernia repair, and are followed for a minimum of 2 years. The target sample size of 753 patients was chosen so that the trial would have power sufficiently high to detect a clinically meaningful difference between treatment groups in either of the two primary outcomes as measured at 2 years: pain or discomfort interfering with normal activities and the physical component summary score of the SF-36 health related quality-of-life survey. The study was begun in five centers located in both community and academic environments. At 18 months, a sixth site was added and at 28 months, after enrollment of 145 patients, one of the centers was terminated for reasons related to inadequate followup; all data from this center were deleted. As a routine measure, an independent experienced trial manager audited all clinical sites. RESULTS: Enrollment of patients began in January 2000 and will end on December 31, 2002. As of November 1, 2002, 637 patients had been randomized, 85% of the target enrollment. An additional 2,115 patients were screened but not randomized, yielding a recruitment rate of 23.1%. Analysis and publication of the results of the study will take place on completion of the minimum 2-year followup period for all patients. CONCLUSIONS: A trial to compare the outcomes of watchful waiting and operation for management of inguinal hernias in men is needed to provide data to surgeons and to patients that can aid in choice of treatment. A description of the design of such a trial is presented. PMID- 12742208 TI - Tension-free inguinal hernia repair: the design of a trial to compare open and laparoscopic surgical techniques. AB - BACKGROUND: Inguinal hernia is a common condition in men and represents a large component of health-care expenditures. Approximately 700,000 herniorrhaphies are performed each year in the United States. The most effective method of repair of an inguinal hernia is not known. STUDY DESIGN: A multicenter, randomized, clinical trial was designed to compare open tension-free inguinal hernia repair with laparoscopic tension-free repair on recurrence rates, complications, patient centered outcomes, and cost. The study design called for randomization of 2,200 men over a period of 3 years. These men will be followed for a minimum of 2 years. This will allow determination of as little as a 3% absolute difference in recurrence rates with 80% power. Randomization is stratified by hospital, whether the hernia is unilateral or bilateral and whether the hernia is primary or recurrent. RESULTS: This is a report of the study design and current status. The study involves 14 Veterans Affairs medical centers with previous experience in laparoscopic hernia repair. After 35 months of enrollment, 2,165 men were randomized and recruitment was then closed. The majority of the patients (82.3%) had unilateral hernias and 90.6% of the hernias were primary. Sixty-seven percent of the patients had an outpatient operation. CONCLUSIONS: We report successful recruitment into a large multicenter trial comparing open and laparoscopic hernia repair. When followup is complete, this study will provide data regarding both clinical (recurrence rates) and patient-centered outcomes. PMID- 12742209 TI - The buccinator musculomucosal island flap for partial tongue reconstruction. AB - BACKGROUND: Ideal tongue reconstruction after partial or total glossectomy should be accomplished with like tissue. The buccinator musculomucosal island flap is similar to lingual tissue, consisting of thin, pliable mucosa of mucus production, with high cell renewal rate and minimal scar formation, excellent color, contour, texture match, and buccinator muscle fibers over the flap's entire length, providing tongual muscle reconstruction without a conspicuous donor site. STUDY DESIGN: The buccinator musculomucosal island flap, based on the facial artery and vein, is designed in a shuttle or in a fish-mouth fashion, encompassing the oral commissure anteriorly. If the flap design is made in a three-leaf shape, a larger flap will be obtained without an oral corner deformity or mouth opening difficulty. The flap is safe and simple to raise. The pedicle of the flap is longer and quite reliable and has a wide range of applicability. The flap may be used for reconstruction of the partial glossectomy defect (tongual defect was not more than half a tongue). The surgeon must know about possible anatomic variations, especially in the venous system, and plan to raise a contralateral buccinator musculomucosal island flap if homolateral facial vascular variation jeopardizes the flap's survival. RESULTS: The flap was successfully used for partial tongue reconstruction in 16 patients, and all flaps have survived without complications. Satisfactory results (including configuration and function of the neotongue) were achieved. Electromyographic studies performed on one patient with half glossectomy revealed reinnervation of the muscle in the flap with active motion of the reconstructed tongue. CONCLUSION: The buccal musculomucosal island flap based on the facial artery and vein is a better reconstruction option with the same or similar kind of tissue as the tongue and, with the addition of the reinnervated flap, offers the potential for improved physiologic motion. PMID- 12742210 TI - Experimental hindlimb ischemia increases neutrophil-mediated matrix metalloproteinase activity: a potential mechanism for lung injury after limb ischemia. AB - BACKGROUND: Acute limb ischemia initiates a systemic inflammatory response, including pulmonary polymorphonuclear leukocyte (PMN) sequestration and acute lung injury. Lung injury is partly attributed to release by PMN's of extracellular matrix (ECM) modifying metalloproteinases (MMPs). We hypothesized that acute hindlimb ischemia (HI) would increase MMP activity in the lung and other organs and that systemic neutrophil depletion before HI would block this effect. STUDY DESIGN: Seventeen FVB/N Tie2/LacZ-182 SATO female mice were randomly divided into four groups: HI + PBS (Group 1), HI + antineutrophil antibody (Group 2), HI + isotype matched control antibody (Group 3), and no HI + PBS (Group 4). HI was achieved by unilateral femoral artery ligation. Neutrophil depletion was confirmed. Three days postligation, lung, liver, and kidney were harvested. MMP-2 and -9 expression and activation (gelatin zymography) and membrane type-1 MMP (MT1-MMP, western blotting) were quantified by densitometry and NIH Image Analysis software. Statistical significance was determined with an analysis of variance. RESULTS: Zymograms revealed a 46% increase in pulmonary proMMP-9 in Group 1 versus Group 4 (6,107 +/- 472 [mean +/- SEM] densitometry units [DU] versus 3,287 +/- 675 DU, p < 0.05). A similar trend was observed for active MMP-9 (3,189 +/- 541 DU versus 1,417 +/- 927 DU, P = 0.16). Neutrophil depletion (Group 2) decreased proMMP-9 levels by 51% (2,996 +/- 314 DU versus 6,107 +/- 472 DU, p < 0.05) and active MMP-9 by 75% (810 +/- 444 DU versus 3,189 +/- 541 DU, p < 0.05) compared with Group 1. Active MMP-2 increased 51% after HI (Group 1, 3,230 +/- 86 DU versus Group 4, 1,599 +/- 327 DU, p < 0.05). Neutrophil depletion decreased the HI-induced activation of MMP-2 by 43% (Group 2, 1,829 +/- 471 DU versus Group 1, 3,230 +/- 86 DU, p < 0.05). CONCLUSIONS: HI increases pulmonary proMMP-9, active MMP-9, and active MMP-2 levels. Neutrophil depletion blocks this effect. These data suggest that acute limb ischemia leads to PMN mediated changes in MMP activity. PMID- 12742211 TI - What's new in cardiac surgery. PMID- 12742212 TI - What's new in general surgery: transplantation. PMID- 12742213 TI - Robotics in surgery. PMID- 12742214 TI - Zuckerkandl's tubercle: Hannibal ad Portas. PMID- 12742216 TI - Robert Wood Johnson Foundation Office of Promoting Excellence in End-of-Life Care: executive summary of the report from the field. PMID- 12742217 TI - Novel technique for control of mediastinal bleeding during thyroidectomy for substernal goiter. PMID- 12742219 TI - Practical and cost effective use of minimally invasive breast biopsy techniques. PMID- 12742220 TI - Extramammary Paget's disease. PMID- 12742222 TI - "Component separation" method for abdominal wall reconstruction. PMID- 12742223 TI - Surgeon volume and carotid endarterectomy. PMID- 12742226 TI - Phospholipase A(2) isoforms: a perspective. AB - Several new PLA(2)s have been identified based on their nucleotide gene sequences. They were classified mainly into three groups: cytosolic PLA(2) (cPLA(2)), secretary PLA(2) (sPLA(2)), and intracellular PLA(2) (iPLA(2)). They differ from each other in terms of substrate specificity, Ca(2+) requirement and lipid modification. The questions that still remain to be addressed are the subcellular localization and differential regulation of the isoforms in various cell types and under different physiological conditions. It is required to identify the downstream events that occur upon PLA(2) activation, particularly target protein or metabolic pathway for liberated arachidonic acid or other fatty acids. Understanding the same will greatly help in the development of potent and specific pharmacological modulators that can be used for basic research and clinical applications. The information of the human and other genomes of PLA(2)s, combined with the use of proteomics and genetically manipulated mouse models of different diseases, will illuminate us about the specific and potentially overlapping roles of individual phospholipases as mediators of physiological and pathological processes. Hopefully, such understanding will enable the development of specific agents aimed at decreasing the potential contribution of individual secretary phospholipases to vascular diseases. The signaling cascades involved in the activation of cPLA(2) by mitogen activated protein kinases (MAPKs) is now evident. It has been demonstrated that p44 MAPK phosphorylates cPLA(2) and increases its activity in cells and tissues. The phosphorylation of cPLA(2) at ser505 occurs before the increase in intracellular Ca(2+) that facilitate the binding of the lipid binding domain of cPLA(2) to phospholipids, promoting its translocation to cellular membranes and AA release. Recently, a negative feed back loop for cPLA(2) activation by MAPK has been proposed. If PLA(2) activation in a given model depends on PKC, PKA, cAMP, or MAPK then inhibition of these phosphorylating enzymes may alter activities of PLA(2) isoforms during cellular injury. Understanding the signaling pathways involved in the activation/deactivation of PLA(2) during cellular injury will point to key events that can be used to prevent the cellular injury. Furthermore, to date, there is limited information available regarding the regulation of iPLA(2) or sPLA(2) by these pathways. PMID- 12742227 TI - Requirement for phosphatidylinositol-3 kinase activity during progression through S-phase and entry into mitosis. AB - Phosphatidylinositol-3 kinase (PI3K) proteins are important regulators of cell survival and proliferation. PI3K-dependent signalling regulates cell proliferation by promoting G1- to S-phase progression during the cell cycle. However, a definitive role for PI3K at other times during the cell cycle is less clear. In these studies, we provide evidence that PI3K activity is required during DNA synthesis (S-phase) and G2-phase of the cell cycle. Inhibition of PI3K with LY294002 at the onset of S-phase caused a 4- to 5-h delay in progression through G2/M. LY294002 treatment at the end of S-phase caused an approximate 2-h delay in progression through G2/M, indicating that PI3K activity functions for both S- and G2-phase progression. The expression of constitutively activated Akt partially reversed the inhibitory effects of LY294002 on mitotic entry, which demonstrated that Akt was one PI3K target that was required during G2/M transitions. Inhibition of PI3K resulted in enhanced susceptibility of G2/M synchronized cells to undergo apoptosis in response to DNA damage as compared to asynchronous cells. Thus, similar to its role in promoting cell survival and cell cycle transitions from G1 to S phase, PI3K activity appears to promote entry into mitosis and protect against cell death during S- and G2-phase progression. PMID- 12742228 TI - Inhibition of Ca(2+) signalling by the sphingosine 1-phosphate receptor S1P(1). AB - The lysophospholipid, sphingosine 1-phosphate (S1P), regulates a multitude of cellular functions by activating specific G protein-coupled receptors (GPCRs) (S1P(1-5), plus three newly identified S1P receptors). The G(i)-coupled S1P(1) receptor inhibits adenylyl cyclase, stimulates mitogen-activated protein kinases (MAP kinases) and cell migration, and is required for blood vessel maturation. Here, we report that S1P(1) inhibits Ca(2+) signalling in a number of cell types. In HEK-293 cells, which endogenously express S1P(1-3), overexpression of S1P(1) reduced intracellular free Ca(2+) concentration ([Ca(2+)](i)) increases induced by various receptor agonists as well as thapsigargin. The inhibitory Ca(2+) signalling of S1P(1) was blocked by pertussis toxin (PTX) and the protein kinase C (PKC) inhibitor, Go6976, and imitated by phorbol ester and overexpression of classical PKC isoforms. Activation of S1P(1) stably expressed in RH7777 cells, which endogenously do not express S1P receptors, also inhibited Ca(2+) signalling, without mediating Ca(2+) mobilization on its own. It is concluded that the widely expressed S1P receptor S1P(1) inhibits Ca(2+) signalling, most likely via G(i) proteins and classical PKC isoforms. Co-expression of S1P(1) with S1P(3), but not S1P(2), reversed the inhibitory effect of S1P(1), furthermore suggesting a specific interplay of S1P receptor subtypes usually found within a single cell type. PMID- 12742229 TI - Histamine potentiates IP(3)-mediated Ca(2+) release via thapsigargin-sensitive Ca(2+) pumps. AB - We have studied the histamine-induced potentiation of inositol 1,4,5 trisphosphate (IP(3))-mediated Ca(2+) release in HeLa cells. Intracellular IP(3) levels were increased by IP(3) dialysis with the whole-cell configuration of the patch-clamp technique (cell dialysis of IP(3)). Low concentrations of extracellular histamine (1 microM) accelerated the rate of IP(3)-mediated Ca(2+) release, an effect that required the coincidence of both histamine signalling and the increase in IP(3) levels. Our data suggest that the potentiation effect of histamine cannot be explained simply by agonist-induced increase in IP(3) levels. Disordering microfilaments with cytochalasin D and microtubules with colchicine caused a decrease in the histamine-induced Ca(2+) response. Furthermore, both cytochalasin D and colchicine diminished the rate of IP(3)-mediated Ca(2+) release, while only the former reduced slightly the histamine-induced potentiation effect. Remarkably, rapid inhibition of SERCA pumps with thapsigargin to avoid the depletion of internal Ca(2+) stores diminished the histamine-induced potentiation of IP(3)-mediated Ca(2+) release, without affecting the rate of IP(3)-mediated Ca(2+) release. These data indicate that histamine-induced potentiation of Ca(2+) release in HeLa cells requires active SERCA pumps and suggest that SERCA pumps are an important factor in determining the efficiency of agonist-induced Ca(2+) release. PMID- 12742230 TI - Growth inhibitory signalling by TGFbeta is blocked in Ras-transformed intestinal epithelial cells at a post-receptor locus. AB - The transforming growth factor beta (TGFbeta) family of growth regulatory peptides plays an important role in the regulation of gastrointestinal epithelial cell homeostasis. Loss of growth inhibitory signalling by TGFbeta is common in the context of Ras-transformation and it has been hypothesized that loss of TGFbeta receptor II (TGFbetaRII) expression accounts for the emergence of TGFbeta resistance. Here we examine the functional significance of reduced TGFbetaRII expression in intestinal epithelial cells transformed by oncogenic Ras. TGFbeta induced signalling events downstream of TGFbetaRII were examined in Ras transformed RIE-1 cells (RIE-Ras) and compared to the parental RIE-1 line. RIE Ras cells were resistant to growth inhibition by TGFbeta. Neither overexpression of TGFbetaRII in RIE-Ras cells nor expression of constitutively active TGFbetaRI restored sensitivity to TGFbeta. TGFbeta-mediated phosphorylation of Smad2 occurred in TGFbeta-resistant RIE-Ras cells, as well as other TGFbeta-resistant cells lines (HT-29, SW620) expressing low levels of TGFbetaRII. Nuclear translocation of Smad2 and Smad4 occurred equally in RIE-Ras and parental RIE cells. The activity of TIEG2, a TGFbeta-inducible SP1-like transcription factor, was reduced in RIE-Ras cells, implying that resistance in Ras-transformed RIE cells occurs by a transcriptional mechanism. PMID- 12742231 TI - SAPK/JNK regulates cdc2/cyclin B kinase through phosphorylation and inhibition of cdc25c. AB - Cells undergo M phase arrest in response to stresses like UV irradiation or DNA damage. Stress-activated protein kinase (SAPK, also known as c-Jun N-terminal kinase, JNK) is activated by such stress stimuli. We addressed the potential effects of SAPK activation on cell cycle regulatory proteins. Activation of SAPK strongly correlated with inhibition of cdc2/cyclin B kinase, an important regulator of G2/M phase. SAPK directly phosphorylated the cdc2 regulator, cdc25c, in vitro on serine 168 (S168). This residue was highly phosphorylated in vivo in response to stress stimuli. cdc25c phosphorylated on S168 in cells lacks phosphatase activity, and expression of a S168A mutant of cdc25c reversed the inhibition of cdc2/cyclin B kinase activity by cell stress. Antibodies directed against phosphorylated S168 detect increased phosphorylation of S168 after cell stress. We conclude that SAPK regulates cdc2/cyclin B kinase following stress events by a novel mechanism involving inhibitory phosphorylation of the cdc2 activating phosphatase cdc25c on S168. PMID- 12742232 TI - Nitrogen-containing bisphosphonates induce S-phase cell cycle arrest and apoptosis of myeloma cells by activating MAPK pathway and inhibiting mevalonate pathway. AB - Bisphosphonates have been used for the treatment of hypercalcemia associated with malignancies and osteoporosis. It was previously reported that the mevalonate pathway is involved in nitrogen-containing bisphosphonate-induced apoptosis in osteoclasts and myeloma cells. The aim of this study was to determine the effects of two bisphosphonates, incadronate, and newly developed bisphosphonate YM529 on human myeloma cells, U266, RPMI-8226, and HS-Sultan. Both incadronate and YM529 induced S-phase cell cycle arrest and apoptosis in these myeloma cells. Treatment of the myeloma cells with cell-permeable substrates for mevalonate pathways, geranylgeraniol, and farnesol prevented bisphosphonate-mediated growth suppression. Checkpoint kinases, Chk1/2, and MAPK became phosphorylated after stimulation with bisphosphonates in the myeloma cells. Bisphosphonate-induced apoptosis was partially prevented by the pretreatment with MAPK inhibitor. These results demonstrate that incadronate and YM529 suppress the proliferation of myeloma cells through mevalonate pathway and MAPK pathway. PMID- 12742234 TI - The ototoxic interaction of styrene and noise. AB - The interaction between noise and inhaled styrene on the structure and function of the auditory organ of the male Wistar rat was studied. The animals were exposed either to 600 ppm, 300 ppm or 100 ppm styrene (12 h/day, 5 days/week, for 4 weeks) alone or in combination with a simultaneous 100-105 dB industrial noise stimulant. Auditory sensitivity was tested by auditory brainstem audiometry at 1.0, 2.0, 4.0 and 8.0 kHz frequencies. Inner ear changes were studied by light microscopy. Exposure to 600 ppm styrene alone caused a 3 dB hearing loss only at the highest test frequency (8 kHz). Quantitative morphological analysis of cochlear hair cells (cytocochleograms) showed a severe outer hair cell (OHC) loss particularly in the third OHC row of the upper basal and lower middle coil. Exposure to noise alone caused only a mild hearing loss (2-9 dB), and only an occasional loss of OHCs (<1% missing). Exposure to the combination of noise and 600 ppm styrene caused a moderate flat hearing loss of 23-27 dB. The cytocochleograms showed a more severe damage of the OHCs than after exposure to 600 ppm styrene alone. The inner hair cells were found to be destroyed in some animals in the upper basal turn only after the combination exposure. Only in combination with noise exposure, the lower styrene concentrations (100 and 300 ppm) induced a hearing loss which was equivalent to that seen after exposure to noise alone. We conclude that: (1) There is an ototoxic interaction between styrene and noise. (2) Synergism is manifested only if styrene is applied in concentrations above the critical level (between 300 and 600 ppm in this study). PMID- 12742233 TI - Effects of alpha-tocopherol on noise-induced hearing loss in guinea pigs. AB - Preventing noise-induced hearing loss (NIHL) by antioxidants is based on the hypothesis that generation of reactive oxygen species is one of the causes of NIHL. alpha-Tocopherol is a naturally occurring antioxidant with no noticeable side effects. In this study, we attempted to protect guinea pigs from developing NIHL by administering alpha-tocopherol. Pigmented male guinea pigs were exposed to a noise (4 kHz octave band, 100 dB SPL), 8 h/day for 3 days consecutively. alpha-Tocopherol (10 mg/kg or 50 mg/kg daily) was given by intraperitoneal injection from 3 days before through 3 days after the noise exposure. Auditory evoked brainstem response (ABR) thresholds at 2, 4 and 8 kHz were recorded prior to the experiment, immediately post-noise, 2 and 8 days post-noise. On day 8 post noise, after the ABR recording, guinea pigs were decapitated and the cochleae were removed for cochlear surface preparations and scanning electron microscope (SEM) study. ABR threshold shifts of groups receiving alpha-tocopherol were significantly smaller than those of groups not receiving alpha-tocopherol at all frequencies and all time points tested except that of group 3 at 8 kHz 8 days post-noise. No hair cell loss was seen on the surface preparations, but stereocilia loss was found by SEM study. The noise-induced stereocilia loss was significantly decreased by alpha-tocopherol. These results indicate that alpha tocopherol can attenuate the noise-induced cochlear damage. Further investigations on the preventive effect of alpha-tocopherol on NIHL in noise exposed workers are necessary. PMID- 12742235 TI - Noise-induced hearing loss in chinchillas pre-treated with glutathione monoethylester and R-PIA. AB - The protective effects of glutathione monoethylester (GEE) and GEE in combination with R-N6-phenylisopropyladenosine (R-PIA) were evaluated in the chinchilla when exposed to impulse (145 dB pSPL) or continuous (105 dB SPL, 4 kHz OB) noise. Six groups of 10 chinchillas were used as subjects. Before exposure to noise, the subjects were anesthetized, a 30 microl drop of drug was placed on the round window (GEE [50, 100, 150 mM], GEE 50 mM and R-PIA). Forty minutes later the subject was exposed to either impulse or continuous noise. The 50 mM treatment provided significant protection from impulse noise, but not from continuous noise exposure. The combination provided significant protection from both the continuous and impulse noise. In a separate set of experiments, glutathione (GSH) levels were measured in the perilymph. All the drug treatments elevated GSH levels. The results are discussed in terms of antioxidant treatments as a prophylactic measure against noise-induced hearing loss. PMID- 12742236 TI - Auditory peripheral influences on calcium binding protein immunoreactivity in the cochlear nucleus during aging in the C57BL/6J mouse. AB - The C57BL/6J (C57) mouse was selected as a suitable model for early presbyacusis to determine if there were correlations between peripheral pathology (spiral ganglion loss, inner and outer hair cell loss) and calcium binding immunoreactivity in the cochlear nucleus during aging. The quantitative stereological method, the optical fractionator, was used for determining the total number of neurons and calcium binding immunopositive neurons (calbindin, parvalbumin and calretinin) during aging in the posteroventral- and dorsal cochlear nucleus (PVCN and DCN) in C57 mice. Comparing 30-month-old to 1-month old C57 mice, a percent increase in parvalbumin and calbindin immunoreactivity was evident in both the PVCN and DCN. Correlations were made between peripheral pathology (spiral ganglion and inner and outer hair cell loss) and calcium binding protein expression. Significant correlations between cochlear pathology and the percentage of parvalbumin and calretinin immunoreactive neurons were demonstrated in the DCN. Moreover, significant correlations were found between cochlear pathology and parvalbumin and calbindin in the PVCN. In summary, the findings imply that degenerative changes in the auditory periphery can modulate neuronal homeostasis by increasing calcium binding proteins in the PVCN and DCN during aging. Taken together, these findings suggest a role for calcium binding proteins in protecting against age-induced calcium toxicity. PMID- 12742237 TI - Establishment and characterization of rat progenitor hair cell lines. AB - Cochlear progenitor hair cell lines are useful for studies of cellular specification, gene expression features, and signal transduction involved in the development of hair cells. To obtain embryonic and postnatal cochlear progenitor hair cell lines, we immortalized primary cultures of sensorineural epithelial cells from otocysts on embryonic day 12 (E12) and explants of the organ of Corti tissues on postnatal day 5 (P5). Primary cultures and explants were then transduced by the E6/E7 genes of human papilloma virus type 16. Transduced cells were passed for >50 passages and partial clonal cells were isolated from the above P5 organ of Corti explants by limiting dilution. The expression of neuronal, neural, epithelial, hair cell markers, and important transcription factors were then examined in these cell clones. Clones that express the above markers were considered as being progenitor hair cells. At least two representative cell lines, one from a mixed culture of otocyst epithelial cells and the other from the organ of Corti cells, ultimately expressed hair cell markers and neuronal/neural cell markers. The former only expressed the early hair cell marker oncomodulin and myosin VIIa, whereas the latter expressed oncomodulin, calretinin, myosin VIIa and Brn 3.1. These cell lines may represent progenitor hair cells at the different stages of cochlear development. PMID- 12742238 TI - Systemic co-treatment with alpha-melanocyte stimulating hormone delays hearing loss caused by local cisplatin administration in guinea pigs. AB - It has previously been demonstrated that ototoxicity induced by systemic administration of cisplatin is reduced by concomitant administration of melanocortins, like alpha-melanocyte stimulating hormone (alpha-MSH). However, these experiments were hampered by large interanimal variability. Therefore, we re-investigated the effects of systemically administered alpha-MSH during local (intracochlear) administration of cisplatin. Guinea pigs, implanted with a round window electrode, allowing daily monitoring of the compound action potentials (CAPs), and a mini-osmotic pump, pumping either 0.5 microl/h physiological saline or cisplatin solution (15 microg/ml), were co-treated daily with a subcutaneous bolus injection of either alpha-MSH (75 microg/kg) or physiological saline for 1 week or until the electrocochleogram showed a persistent decrease in CAP amplitude (40 dB threshold shift at 8 kHz). Next, the animals were sacrificed and the cochleas were processed for histology. After 2-3 days, cisplatin alone caused a threshold shift at all frequencies (2-16 kHz). Co-administration with alpha-MSH consistently delayed the criterion threshold shift by 1 day. When the 40 dB criterion had been reached, similar outer hair cell losses in both the cisplatin/alpha-MSH- and cisplatin/saline-treated groups were observed. This experiment confirms that direct administration of cisplatin into the cochlea results in considerably less interanimal variability than systemic administration and that co-treatment with alpha-MSH delays cisplatin ototoxicity. Since cisplatin was delivered directly to the cochlea, the ameliorating effect of alpha MSH probably involves a cochlear target. PMID- 12742239 TI - Single electrode micro-stimulation of rat auditory cortex: an evaluation of behavioral performance. AB - A combination of electrophysiological mapping, behavioral analysis and cortical micro-stimulation was used to explore the interrelation between the auditory cortex and behavior in the adult rat. Auditory discriminations were evaluated in eight rats trained to discriminate the presence or absence of a 75 dB pure tone stimulus. A probe trial technique was used to obtain intensity generalization gradients that described response probabilities to mid-level tones between 0 and 75 dB. The same rats were then chronically implanted in the auditory cortex with a 16 or 32 channel tungsten microwire electrode array. Implanted animals were then trained to discriminate the presence of single electrode micro-stimulation of magnitude 90 microA (22.5 nC/phase). Intensity generalization gradients were created to obtain the response probabilities to mid-level current magnitudes ranging from 0 to 90 microA on 36 different electrodes in six of the eight rats. The 50% point (the current level resulting in 50% detections) varied from 16.7 to 69.2 microA, with an overall mean of 42.4 (+/-8.1) microA across all single electrodes. Cortical micro-stimulation induced sensory-evoked behavior with similar characteristics as normal auditory stimuli. The results highlight the importance of the auditory cortex in a discrimination task and suggest that micro stimulation of the auditory cortex might be an effective means for a graded information transfer of auditory information directly to the brain as part of a cortical auditory prosthesis. PMID- 12742241 TI - Hyperthermia exacerbates and hypothermia protects from noise-induced threshold elevation of the cochlear nerve envelope response in the C57BL/6J mouse. AB - The scalp-recorded cochlear nerve envelope response (CNER) reflects the ability of high frequency cochlear nerve axons to fire in a phase-locked fashion to low frequency modulations of the acoustic envelope of high frequency stimuli. This property might be useful in evaluating the adverse effects of noise exposure on the ability of the ear to detect acoustic changes characteristic of vocalizations and speech. Hyperthermia (40 degrees C rectal) per se had no observable influence on the CNER in C57BL/6 mice. Hypothermia (30 degrees C) elevated CNER thresholds elicited by high frequency stimuli, although these stimuli still generated an auditory brainstem response. Mice exposed to noise when hyperthermic had greater threshold elevations than those exposed when euthermic (36 degrees C); those exposed when hypothermic had smaller threshold elevations than those exposed when euthermic. These observations were discussed in terms of the interaction of temperature and noise on oxidative processes within the cochlea. PMID- 12742240 TI - Spatial spread of neural excitation in cochlear implant recipients: comparison of improved ECAP method and psychophysical forward masking. AB - This study introduces and evaluates a method for measurement of the longitudinal spread of electrically evoked neural excitation in the cochlea, using the Neural Response Telemetry system (NRT) available with the Nucleus((R)) 24 cochlear implant system. The recently released version of the NRT software (version 3.0) enables presentation of the 'masker' and 'probe' on different electrodes. In the present method the probe position was fixed, while the masker position was varied across the electrode array. The amplitude of the response to the partially masked probe provides a measure of the amount of masking, which is dependent on the extent of overlap of the excitation regions of the masker and probe. These measurements were performed in seven subjects implanted with the Nucleus 24 cochlear implant system (four with straight and three with Contour electrode arrays), for basal, middle and apical probe electrodes. Similar excitation profiles were obtained using either the standard NRT subtraction paradigm or an alternative 'Miller' method. The excitation profiles were compared with those obtained from psychophysical forward masking and good agreement was found. The widths of electrically evoked compound action potential (ECAP) and forward masking profiles did not differ significantly. Whereas the width of the ECAP measure was significantly correlated with both the maximum comfortable level and the distance of the electrode band from the modiolus, the width of the forward masking profile was not. PMID- 12742242 TI - The interaction between ear and sex differences and stimulus rate. AB - Evoked otoacoustic emissions (EOAEs) are produced by the cochlea in response to acoustic stimuli and provide an objective and non-invasive measure of cochlear function. A new technique, based on maximum length sequences (MLSs), enables stimulus rates of up to 5000 clicks/s to be used. Conventional EOAE amplitude differs between ears and sexes, female subjects having responses of greater amplitude than male subjects and right ears larger responses than left ears. As a prerequisite to clinical use it is necessary to establish if these differences occur with the MLS OAE technique and whether they change with stimulus rate. Eighty ears of normally hearing adults between the ages of 18 and 40 years were tested. MLS OAEs were recorded at eight stimulus rates ranging from 40/s to 5000/s. Two stimulus levels and two recordings were made at each stimulus rate. Female subjects were found to have statistically significantly larger MLS OAEs than male subjects and gave larger amplitude responses in the right ears. The difference was not significant between male right and left ears. A rate effect was also demonstrated with the amplitude of the MLS OAEs decreasing with an increase in rate. The study provides normative data for MLS OAE testing and shows that females have MLS OAEs of larger amplitude than males and that as the click stimulus rate increases the significance of this difference decreases. Female right ears also have MLS OAEs of greater amplitude than female left ears. PMID- 12742243 TI - The sound-level-dependent growth in the extent of fMRI activation in Heschl's gyrus is different for low- and high-frequency tones. AB - fMRI (functional magnetic resonance imaging) was used to investigate whether the growth in activation of the human auditory cortex, with increasing sound level, is discernibly different for high- and low-frequency tones. Ten volunteers were scanned whilst listening to sequences of low-frequency (0.30-kHz) tones at sound levels between 42 and 96 dB sound pressure level (SPL), and 10 whilst listening to high-frequency (4.75-kHz) tones at the same sound levels. Activation was measured in Heschl's gyrus (including primary auditory cortex) which has been shown to be most sensitive to changes in sound level. For the 0.30-kHz tone, the extent of activation was flat up to 66 dB and then showed a rapid growth which continued up to the highest level studied (96 dB SPL). In contrast, increasing the level of 4.75-kHz tones produced a steady growth in the extent of activation across the range of levels studied. These results are consistent with physiological evidence suggesting that recruitment of primary auditory cortical neurones may be different at high and low frequencies. PMID- 12742244 TI - Properties of Volterra slices of otoacoustic emissions in normal-hearing humans obtained using maximum length sequences of clicks. AB - Nonlinear temporal interaction components of otoacoustic emissions (OAEs) may be investigated by presenting a stream of clicks in maximum length sequences. This yields responses, termed here Volterra slices, which are related to the Volterra kernels of the system. The aim of this study was to obtain normative data on Volterra slices over a range of click rates and stimulus levels. OAEs were recorded in 12 normally hearing adult ears at six rates and four click levels. In addition to the first order kernel, six slices from the Volterra slices of orders 2-5 were extracted from the recordings. It was found that higher order kernel slices could be reliably measured in all 12 ears tested and that they have properties that differ from those of the conventional OAEs. These findings may facilitate the study of cochlear function in both normal and pathological ears. PMID- 12742245 TI - Partial reinforcement across trials impairs escape performance but spares place learning in the water maze. AB - We studied the effects of partial reinforcement on escape performance and place learning in the water maze. Rats given 50% reinforcement across trials (i.e. the escape platform was present only on odd trials) were compared to controls given 100% reinforcement (platform present on all trials). Control groups either received 8 or 4 trials per day, which was equal to either the total number of trials (100%-8) or reinforced escapes (100%-4) of the 50% group. Analysis of escape performance (latency) revealed that the 50% group was impaired relative to the 100%-8 group, but not the 100%-4 group, during the first 5 days of acquisition. The 50% group was impaired relative to both control groups on days 6 10 of overtraining. However, analyses of within-trial behavior (target annulus preference and thigmotaxis) on nonreinforced trials suggest that the 50% group did learn the location of the hidden platform (place information), in addition to a wall-based thigmotactic response. By dividing the 60s nonreinforced trials into three 20-s time bins, we were able to detect a significant preference for the target annulus early in the trial (bin 1 of trial 40 and bins 1-2 of trial 80). Further, there was a significant increase in time spent in the periphery of the pool, near the wall, in the last time bin of trial 40. Because the platform was in the middle zone, this behavior competed with a place response. We conclude that across-trial partial reinforcement procedures may promote response competition and mask evidence of place learning in addition to weakening escape performance late in training. PMID- 12742246 TI - Behavioral effects of neonatal and adult excitotoxic lesions of the mediodorsal thalamus in the adult rat. AB - We examined in the rat, the effects of neonatal (postnatal Day 7) and adult excitotoxic lesions of the mediodorsal thalamus (MDT), a brain area innervating the prefrontal cortex and implicated as a site of neuropathology in schizophrenia. Previous studies showed that rats with neonatal excitotoxic damage of the ventral hippocampus (VH), used as an animal model of this disorder, display in young adulthood a variety of abnormalities reminiscent of schizophrenia, including hyperactivity to stressful stimuli and amphetamine. It has been speculated that behavioral abnormalities of the neonatally VH lesioned animals are mediated through MDT projections to the prefrontal cortex. We tested if rats with ibotenic acid (1.5 microg per hemisphere in neonates, 2 microg in adults) lesions of MDT exhibited motor hyperactivity in the same experimental conditions (i.e. in response to novelty, saline injections and amphetamine administration) as rats with the VH lesions. We found that, in contrast to rats with VH lesions, neonatally lesioned MDT rats showed reduced vertical activity in response to amphetamine and no changes in locomotor activity to novelty, saline or amphetamine injections 7 weeks postlesion. Adult lesioned MDT rats exhibited no changes in motor activity as compared to controls at 7 weeks postlesion. These results indicate that neonatal or adult excitotoxic lesions of MDT do not produce behavioral changes analogous to those seen after neonatal VH lesions and do not appear to reproduce animal model-like features of schizophrenia. PMID- 12742247 TI - The effect of isolation rearing on volitional ethanol consumption and central CCK/dopamine systems in Fawn-Hooded rats. AB - Numerous studies have demonstrated that socially isolating rats (from weaning) produces a sustained anxious phenotype and an enhanced response to psychostimulant drugs such as amphetamine and cocaine. In addition, isolation rearing has been shown to induce significant changes in the mesolimbic dopamine system. These data indicate that isolation rearing not only induces an anxiogenic phenotype but also induces neurochemical changes in reward nuclei of the brain, which is correlated with an enhanced response to psychostimulants. For these reasons, the effect of isolation rearing on volitional ethanol consumption was examined in Fawn-Hooded (FH) rats and correlated with neurochemical changes in central dopamine and cholecystokinin systems. Social isolation from weaning produced an anxiogenic phenotype as measured by a decreased time spent on the open arms of an elevated plus-maze. Interestingly, isolation-rearing induced a greater proportion of FH rats to acquire preference for ethanol while having no effect on the amount of ethanol consumed by alcohol-preferring rats. In addition, isolation rearing induced a number of changes in central CCK/dopamine systems. PMID- 12742248 TI - High frequency (200 Hz) oscillations and firing patterns in the basolateral amygdala and dorsal endopiriform nucleus of the behaving rat. AB - The known repertoire of rhythms in the amygdala and paleocortex includes a range of oscillations from slow waves (<1 Hz) to fast gamma (40-100 Hz). In the present report, we show approximately 200 Hz oscillations in the basolateral nucleus of the amygdala (BL) and the adjacent dorsal endopiriform nucleus (EPN) of the behaving rat. Microwire techniques were applied for recording single units and field activity from these structures and EEG from the dorsal or temporal CA1 subfields of the hippocampus. Units from both EPN and BL exhibited similar irregular firing patterns with bursts. The mean firing rates in EPN were <1 Hz, whereas units in the BL fired in a range of <1-17 Hz. Neuronal activity in both BL and EPN was phase-locked with high-frequency field oscillations (HFO, approximately 200 Hz). Amygdaloid/EPN HFO displayed on average lower numbers of cycles and smaller amplitudes than hippocampal ripples. Neuronal firing and HFO in the BL and EPN were state dependent with a maximal occurrence during slow-wave sleep (SWS), being lower during waking and paradoxical sleep. Cross-correlation between hippocampal ripples and EPN or BL units and field HFO did not reveal any synchrony. These data suggest common principles of temporal coding in BL and EPN in certain behavioural states via short scale population synchrony though they convey signals of different modalities. PMID- 12742249 TI - Lateralization of aggression in fish. AB - Recent research has suggested that lateralization of aggressive behaviors could follow an homogeneous pattern among all vertebrates. A left eye/right hemisphere dominance in eliciting aggressive responses has been demonstrated for all groups of tetrapods but teleost fish for which data is lacking. Here we studied differential eye use during aggressive interactions in three species of teleosts: Gambusia holbrooki, Xenotoca eiseni and Betta splendens. In the first experiment we checked for lateralization in the use of the eyes while the subject was attacking its own mirror image. In order to confirm the results, other tests were performed on two species and eye preference was scored during attacks or displays directed toward a live rival. All three species showed a marked preference for using the right eye when attacking a mirror image or a live rival. Thus, the direction of asymmetry in fish appears the opposite to that shown by all the other groups of vertebrates. Hypotheses on the origin of the difference are discussed. PMID- 12742250 TI - Operant learning and differential-reinforcement-of-low-rate 36-s responding in 5 HT1A and 5-HT1B receptor knockout mice. AB - Previous studies with mice lacking 5-HT(1A) (1AKO) and 5-HT(1B) (1BKO) receptors in hippocampus-dependent learning and memory paradigms, suggest that these receptors play an important role in learning and memory, although their precise role is unclear. In the present study, 1AKO and 1BKO mice were studied in operant behavioural paradigms of decision making and response inhibition, to further study the putative involvement of these receptors in prefrontal cortex-dependent learning and memory. Moreover, because 1AKO mice have been shown to exhibit an antidepressant-like phenotype and 1BKO mice to be more impulsive in ethological studies, mice were trained in a differential-reinforcement-of-low-rates (DRL) procedure. Overall, results indicate that 1AKO and 1BKO mice display subtle differences in operant paradigms of decision making and response inhibition compared to wild type (WT) mice. In addition, when responding under a DRL 36-s schedule had stabilised, 1BKO mice showed a phenotype indicative of increased impulsivity, whereas 1AKO mice did not differ from WT mice. In conclusion, 5 HT(1B) receptors appear to play an important role in impulsivity and a minor role in prefrontal cortex-dependent learning and memory as shown by the results obtained in serial reversal learning and extinction. In contrast, 5-HT(1A) receptors appear to be involved in facilitation of autoshaping, but their role in impulsivity and prefrontal cortex-dependent learning and memory appears to be limited. PMID- 12742251 TI - Improvement of shuttle-box performance by anterodorsal medial septal lesions in rats. AB - Septal lesions impair a variety of tasks, including inhibitory avoidance and one way active avoidance. In contrast, these lesions improve two-way active avoidance, probably by reducing anxiety. The present work aimed to study whether anterodorsal medial septal lesion (a) improves performance of two-way active avoidance task (Experiment I), as it has been observed with wider septal lesion, and (b) affect anxiety and/or locomotor activity (Experiment II). This precise region was chosen because some evidences suggest that its lesion do not lead to a reduction of anxiety. Lesioned rats tended to make a higher, but statistically non-significant (P=0.074), number of avoidances regardless of the session, being this difference statistically significant on the retention session (RT). The same lesion did not appear to have an anxiolytic effect, and did not affected basal locomotor activity. Different possible explanations of our results are discussed. PMID- 12742252 TI - Instrumental learning within the spinal cord: V. Evidence the behavioral deficit observed after noncontingent nociceptive stimulation reflects an intraspinal modification. AB - Spinally transected rats given leg shock whenever one hindlimb is extended learn to maintain the leg in a flexed position, which minimizes net shock exposure. Yoked rats, that receive an equal amount of shock independent of leg position (noncontingent shock), do not exhibit an increase in flexion duration. Yoked rats also fail to learn when response contingent shock is applied to the previously shocked leg, a behavioral deficit that resembles learned helplessness. This deficit could reflect either a peripheral (e.g. muscle fatigue) or central effect. Experiment 1 showed that spinalized rats given noncontingent shock to one hind limb fail to learn when response-contingent shock is applied to the contralateral leg. Experiment 2 demonstrated that blocking the afferent input to the spinal cord, by cutting the sciatic nerve, blocked the development of the deficit. Experiment 3 found that intrathecal lidocaine has a protective effect and prevents the deficit. These findings suggest that noncontingent nociceptive stimulation induces an intraspinal modification that undermines behavioral potential. PMID- 12742253 TI - Removal of short-term isolation stress differentially influences prepulse inhibition in APO-SUS and APO-UNSUS rats. AB - Epidemiological studies have reported that the risk of developing schizophrenia increases with the number of genes one shares with patients suffering from schizophrenia [Gottesman Schizophrenia Genesis, New York: Freeman; 1991]. In addition, stressful life events are known to increase the risk of developing schizophrenia [Schizophr Res 30 (1998) 251] resulting in the stress hypothesis of schizophrenia. Remarkably, stress increases the release of dopamine and noradrenaline in the nucleus accumbens [Brain Res 554 (1991) 217], which links the stress hypothesis with the known dopamine hypothesis of schizophrenia. Additionally an increased dopamine transmission in the nucleus accumbens (Nacc) is known to disturb prepulse inhibition (ppi) [Pharmacol Biochem Behav 49 (1994) 155], a phenomenon observed in, among others, schizophrenics [Arch Gen Psychiatry 47 (1990) 181]. Some years ago we have genetically selected two rat-lines which are marked by a high (APO-SUS) and by a low (APO-UNSUS) apomorphine susceptibility. Similar to schizophrenics the APO-SUS rat-line shows a reduced ppi [J Neurosci 15 (1995) 7604]. However, these data were obtained after a period of mild stress, namely a 24-h period of social isolation. Mild stress changes the line specific differences of APO-SUS and APO-UNSUS rats. The stress pushes the APO-SUS rat in the direction of an APO-UNSUS and vice versa, especially as far as it concerns the dopamine and noradrenaline activity in the nucleus accumbens [Cools AR, van-den Bos R, Ellenbroek BA, Gaiting function of noradrenaline in the ventral striatum: its role in behavioural responses to environmental and pharmacological challenges. In: Willner P, Scheel-Kruger J, editors. The mesolimbic dopamine system: from motivation to action. New York: Wiley; 1991 [Chapter 6]; Cools AR, Rots NY, De-Kloet ER, Apomorphine-susceptible and apomorphine-unsusceptible Wistar rats: a new tool in the search for the function of striatum in switching behavioural strategies. In: Pea G (Ed.), The basal ganglia IV, New York: Plenum Press; 1994; Brain Res Bull 24 (1990) 49; Behav Neurosci 108 (1994) 1107]. Therefore, in the present paper we investigated the ppi response in non-stressed, i.e. non-isolated APO-SUS and APO-UNSUS rats. In agreement with this hypothesis, we found that removal of the stress led to an increase of ppi in the APO-SUS, but a decrease in the APO-UNSUS. These data clearly shows that the ppi is stress-dependent in APO-SUS and APO-UNSUS rats. It is suggested that the differential stress-induced change in the dopaminergic and the noradrenergic system influences the reaction of APO-SUS and APO-UNSUS rats on ppi. PMID- 12742254 TI - MPEP, a selective metabotropic glutamate receptor 5 antagonist, attenuates conditioned taste aversion in rats. AB - Metabotropic glutamate receptors (mGluRs) have been implicated in several types of cognitive and associative learning. Although recent evidence indicates an influence of mGluRs in conditioned taste aversion (CTA), the subtype-specific involvement of mGluRs in this learning paradigm remained to be determined. The aim of this study was to examine the role of Group I mGluR subtypes in CTA using a selective mGluR5 antagonist (2-methyl-6-(phenylethynyl)-pyridine, MPEP) and a selective mGluR1 antagonist (1-aminoindan-1,5-dicarboxylic acid, AIDA). Male, water-deprived, Sprague-Dawley rats were injected i.p. with 6 or 12 mg/kg MPEP or saline. Twenty-five minutes later, all rats received 15-min access to a 0.1% saccharin solution (Sac) immediately followed by an injection of 0.15M LiCl at 1.33% body weight. The animals were tested with 15-min access to Sac on each of four test days. MPEP-treated animals consumed more Sac on the test trials than saline-treated rats. In another experiment, controlled access to Sac was used by infusing the solution on the conditioning trial. Consistent with the above results, MPEP attenuated the degree of CTA. Similar experiments using the mGluR1 antagonist AIDA, have found no effect on CTA learning. These results suggest that the two subtypes of Group I mGluRs are differentially involved in taste aversion learning. PMID- 12742255 TI - Perirhinal cortex lesions produce variable patterns of retrograde amnesia in rats. AB - Two experiments examined the contribution of the perirhinal cortex (PRh) to retrograde memory for the location of a platform in a water maze. In a previous study, we found that electrolytic lesions of the PRh produced retrograde amnesia, without a temporal gradient, for water-maze problems acquired 4 weeks and 2 days before surgery [Behav. Brain. Res. 114 (2000) 119]. In Experiment 1, we used the same mixed design as in our previous report (time of learning was a within subjects factor), but PRh lesions were made by aspiration. Contrary to our earlier report, these PRh rats displayed good retention of both platform locations. Combined, these findings indicate that the lesion method may contribute importantly to the pattern of deficits observed. Experiment 2 was conducted similar to Experiment 1, except that a completely between-subjects design was used (time of learning was a between-subjects factor). Rats that received PRh lesions approximately 2 days after the last training session displayed impaired retention of the platform's location, whereas rats that received PRh lesions 4 weeks after training did not. This finding of a temporally graded retrograde amnesia is consistent with our earlier report, and further suggests that the involvement of the PRh in the retention of water-maze problems is time-limited. However, also consistent with our earlier report, the PRh lesioned rats in Experiment 2 that displayed a retention deficit rapidly reacquired the task. This finding, combined with the negative findings in Experiment 1, suggests that the contribution of the PRh to retrograde memory for platform locations is subtle and may not be due to impaired spatial memory abilities. Additionally, the conflicting results of Experiments 1 and 2 underscore the importance of the design employed in studies of retrograde amnesia in animals. PMID- 12742256 TI - Postnatal hypobaric hypoxia in rats impairs water maze learning and the morphology of neurones and macroglia in cortex and hippocampus. AB - Newborn rats were exposed to intermittent hypobaric hypoxia from birth until the age of 19 days. Spatial memory was tested in a Morris water maze from postnatal day (P) 23 to P32 and from P100 to P109. From P24 to P27 and on days P100 and P101, the escape latencies of hypoxic animals were longer than those of controls. At P24, the number of neuronal bodies increased in cortical layer II of the somatosensory, motor, and auditory areas, and in layer V of the motor area, but the number of neuronal bodies throughout the whole cortical thickness was unchanged. Decreases in the immunostaining density for neurofilaments (anti-NF 160), astrocytes (anti-GFAP), and oligodendrocytes (RIP) were found in the hippocampus, and the typical parallel organisation of neuronal and macroglial processes was lost. Decreases in immunostaining for neurofilaments and oligodendrocytes were also found in the somatosensory cortex and motor cortex. In adult hypoxic rats, at P114-P240, the number of neuronal bodies and the immunostaining density for neurofilaments, astrocytes, and oligodendrocytes in the examined areas were similar to adult controls; however, in the hippocampus we found hypertrophy of fine astrocytic processes and a decreased number of oligodendrocytic processes. We conclude that the neonatal brain damage induced by hypobaric hypoxia impairs spatial memory in infant as well as adult rats. Hypobaric hypoxia delays the maturation of neurones and substantially affects macroglia in the cortex and hippocampus. PMID- 12742257 TI - Hypothermia in mice tested in Morris water maze. AB - The Morris water maze, one of the most common behavioral tasks to assess learning and memory in rodents, exposes the animals to cold water for a few minutes. Unlike rats, young healthy mice can become severely hypothermic during the task. Five swims of 45 s in 20 degrees C water with 30s between the trials was enough to cause up to 9 degrees C drop in the rectal temperature. The decline in core temperature was accompanied by slowing of the swimming speed. Moreover, the effect was dependent on the sex and genotype of the mice, such that females were more susceptible to hypothermia than males and transgenic mice carrying Alzheimer associated APP and PS1 mutations more vulnerable than their nontransgenic littermates. Raising the water temperature from 20 to 24 degrees C alleviated the hypothermia, but did not remove the significant drop in core temperature when using 30-s inter-trial interval. However, increasing the break from 30 s to 13 min removed the net cooling effect of five trials on the core temperature and swimming speed. We conclude that the currently most common water maze protocol renders mice hypothermic, which may confound the test results, especially when transgenic female mice are used. We recommend monitoring of the swimming speed on a trial-by-trial basis and using longer inter-trial intervals. PMID- 12742258 TI - WIN 55,212-2 decreases the reinforcing actions of cocaine through CB1 cannabinoid receptor stimulation. AB - CB(1) cannabinoid receptor agonists show a different profile compared to other drugs of abuse on the basis of experimental data that reveal their reinforcing properties. Thus, there are controversial data in the literature concerning the ability of CB(1) receptor agonists to reinforce behavioral responses in experimental animals, i.e. to lower self-stimulation thresholds, and to support self-administration or conditioned place preference. The aim of the present study was to examine the effects of WIN 55,212-2, a potent CB(1) receptor agonist (graded doses 0.1, 0.3, 1 mg/kg, i.p.), on the rewarding efficacy of lateral hypothalamic self-stimulation and on the systemic cocaine-induced potentiation of brain-stimulation reward. WIN 55,212-2 did not affect lateral hypothalamic self stimulation thresholds both in drug nai;ve rats and in rats pretreated with the drug, whereas it produced a significant, dose-dependent decrease in the maximal rate of responding, i.e. in the performance of the animals. Cocaine (5.0 mg/kg, i.p.) produced a significant reduction in self-stimulation threshold, without altering maximal rates of responding. Importantly, WIN 55,212-2 attenuated the effect of cocaine at the two higher doses tested. The effects of the CB(1) receptor agonist were reversed by pretreatment with the selective CB(1) receptor antagonist SR 141716A (0.02 mg/kg, i.p.) that did not by itself affect cocaine's action. These results indicate that acute stimulation of CB(1) receptors per se does not affect baseline self-stimulation, but reduces the reinforcing effects induced by cocaine. Taken together these findings suggest that cannabinoids may interfere with brain-reward systems responsible for the expression of acute reinforcing properties of drugs of abuse, such as cocaine, and provide evidence that the cannabinoid system could be an interesting drug discovery and development target for the treatment of drug addiction. PMID- 12742259 TI - Subhypnotic doses of propofol accelerate extinction of conditioned taste aversion. AB - Subhypnotic doses of propofol accelerate extinction of conditioned taste aversion. Some intravenous anesthetic agents including propofol is known to induce anterograde and retrograde amnesia. We evaluated whether propofol affect the long-term memory formed by the conditioned taste aversion (CTA) paradigm. Rats were allowed a 4h access to water through the experiments. After preconditioning water intake, the rats were offered 0.1% sodium saccharin (Sac) as conditioned stimulus (CS) for 20 min. An intraperitoneal (i.p.) injection of several concentrations (0.5-100 mg/kg) of propofol 10 min after Sac exposure was followed by an i.p. injection of 0.15M LiCl (2% of body weight) as unconditioned stimulus (US) 30 min after CS-exposure. The volumes of intake of Sac for 20 min were measured on the successive 4 days. The rats, which acquired CTA by every CS US paradigm, strongly avoided Sac on the first test day after conditioning and maintained the avoidance for 3 days. However, when subhypnotic dose of propofol was injected before LiCl-injection, Sac intake abruptly increased on the second test day and the almost complete extinction occurred on the third test day after conditioning. The extinction process of CTA was barely affected by hypnotic dose of propofol. These results suggest that propofol affects the retention mechanism of the CTA memory in a dose-dependent manner. Subhypnotic dose of propofol may affect the sub-cellular process of the memory consolidation in CTA. PMID- 12742260 TI - Characterization of social behaviors and oxytocinergic neurons in the S-100 beta overexpressing mouse model of Down Syndrome. AB - S-100 beta, a gene triplicated in Down Syndrome (DS), is thought to play a role in development of the brain in general, and in the serotonergic neuronal system in particular. We have been studying an animal model of DS, based on overexpression of this gene. In the current study, we report on the social behaviors of these animals, both in same-strain and mixed-strain pairings. In addition, as the neuropeptide oxytocin is often thought to be involved in social behaviors, we have looked at oxytocin-containing cells. In non-social behaviors, such as grooming and line-crossing, the S-100 beta animals were more active than the CD-1 control animals and showed significantly less social sniffing. In mixed strain studies, these differences became more pronounced, with the CD-1 animals showing significantly greater levels of sniffing and anogenital sniffing. As well, the CD-1 animals showed more rearing and an increase in line crossings, suggesting a heightened level of vigilance or awareness of novelty. The S-100 beta animals, conversely, did not appear to respond to the novelty of the CD-1 animals. In mixed pair studies, the S-100 beta animals more frequently took submissive postures, while the CD-1 animals more frequently took dominant postures, and showed a significant increase in biting the S-100 beta partner. The S-100 beta animals showed less rearing, perhaps a further indication that they were inhibited by the CD-1 animals. Analysis of oxytocin-containing neurons showed comparable levels in the supraoptic and paraventricular nuclei of the hypothalamus, but significantly reduced numbers of cells in the bed nucleus of the stria terminalis of the S-100 beta animals. These results are discussed in terms of oxytocin contributions to socialization and fear responding and the significance of these findings to DS. PMID- 12742261 TI - Influence of task parameters on rotarod performance and sensitivity to ethanol in mice. AB - Motor performance in mice can be assessed with multiple apparatus and protocols. Use of the rotarod (a.k.a. rotorod, rota-rod, roto-rod, or accelerod) is very common, and it is often used with the apparent assumption by the experimenters that it is a straightforward and simple assay of coordination. The rotarod is sensitive to drugs that affect motor coordination, including ethanol. However, there are few systematic data assessing the range of "normal" performance in mice. There are also few data exploring optimal task parameters (e.g. the influence of different speeds of rotation). In these experiments, we show that both accelerating and fixed-speed rotarod (FSRR) performance vary under different test protocols and conditions, and that moderate to high doses of ethanol disrupt performance. Under certain conditions, low doses of ethanol were found to enhance performance on the accelerating rotarod (ARR). Therefore, it is not possible to characterize individual differences fully using a single set of test parameters. For example, because of the biphasic effect of ethanol on performance, at least two doses of the drug are necessary to explore individual sensitivity differences. We offer recommendations of parameters we believe to be generally suitable for exploring the performance of new genotypes using the rotarod. We suggest that other putative tests of "ataxia" are similarly complex, and that characterizing the contribution of genetic differences will require similar attention to the details of task apparatus and protocols. These data also underscore the need to employ multiple behavioral assays in order to model a complex domain such as "ataxia" or "coordination." PMID- 12742262 TI - Neuropeptide Y (NPY) Y2 receptors mediate behaviour in two animal models of anxiety: evidence from Y2 receptor knockout mice. AB - The behavioural phenotype of mice lacking neuropeptide Y (NPY) Y(2)-type receptors was assessed in two well documented animal models of anxiety: namely, the elevated plus maze and the open field. NPY Y(2)-/- mice made more entries into, and spent significantly more time on, the open arms of the elevated plus maze when compared to their wild-type Y(2)+/+ controls (P<0.001). This effect was not due to non-specific changes in locomotor activity as the number of closed arm entries did not differ between groups. In addition, NPY Y(2)-/- mice displayed increased preference for the central area of the open field when compared to Y(2)+/+ animals (P<0.01), whereas total entries did not differ between groups. This study suggests that NPY Y(2) receptors may play an inhibitory role and supports the hypothesis that Y(2) receptors are involved in the regulation of anxiety-like behaviours by NPY. PMID- 12742263 TI - Complexity and simplicity in the measurement and recording of the adverse effects of cancer treatment. PMID- 12742264 TI - Mucositis incidence, severity and associated outcomes in patients with head and neck cancer receiving radiotherapy with or without chemotherapy: a systematic literature review. AB - BACKGROUND AND PURPOSE: To determine the frequency of mucositis and associated outcomes in patients receiving radiotherapy (RT) for head and neck cancer through a systematic review of recently published literature. MATERIALS AND METHODS: According to the study protocol, databases were searched for randomized clinical trials (English only, 1996-1999) of patients with head and neck cancer receiving RT with or without chemotherapy that reported one or more outcomes of interest. RESULTS: Thirty-three studies (n=6181 patients) met inclusion criteria. Mucositis was defined using a variety of scoring systems. The mean incidence was 80%. Over one-half of patients (56%) who received altered fractionation RT (RT-AF) experienced severe mucositis (grades 3-4) compared to 34% of patients who received conventional RT. Rates of hospitalization due to mucositis, reported in three studies (n=700), were 16% overall and 32% for RT-AF patients. Eleven percent of patients had RT regimens interrupted or modified because of mucositis in five studies (n=1267) reporting this outcome. Data insufficiency or heterogeneity prohibited analysis of mucositis severity and other associated outcomes, such as oral pain, dysphagia and opioid use. CONCLUSIONS: Mucositis is a frequent, severe toxicity in patients treated with RT for head and neck cancer. While it appears that mucositis may lead to hospitalization and treatment interruptions, its overall impact on outcomes has not been adequately investigated. PMID- 12742266 TI - Waiting times for radiotherapy: consequences of volume increase for the TCP in oropharyngeal carcinoma. AB - BACKGROUND AND PURPOSE: Waiting lists for radiotherapy have become longer over the past years. Apart from the psychological distress for the patient we are concerned about tumour growth during this waiting time, which may worsen prognosis. The purpose of this pilot study was to investigate tumour growth in the waiting time and to obtain an indication of its clinical consequences for patients with oropharyngeal carcinoma. A tumour control probability (TCP) model was applied to evaluate consequences for outcome. METHODS AND MATERIALS: Increase in tumour volume was measured for 13 patients with oropharyngeal carcinoma by outlining the tumour on the diagnostic as well as on the treatment planning CT scan. Waiting time was defined as time between histopathological diagnosis and start of radiotherapy. For each tumour we calculated the increase in tumour volume and the tumour doubling time. The potential increase in TCP was calculated for each tumour for the situation without treatment delay. RESULTS: The mean increase in tumour volume was 70%. The mean waiting time was 56 days. Expected TCP with incorporation of delay was 47%, without delay it might have been 63-66%. CONCLUSION: This study shows tumour progression during the time between the diagnostic CT scan and the treatment planning CT scan in oropharyngeal cancer. As a consequence of waiting time, which allows tumour volume increase, there may be an average control loss of 16-19 % for these tumours during the total waiting time before radiotherapy. PMID- 12742265 TI - A study of the effects of internal organ motion on dose escalation in conformal prostate treatments. AB - BACKGROUND AND PURPOSE: To assess the effect of internal organ motion on the dose distributions and biological indices for the target and non-target organs for three different conformal prostate treatment techniques. MATERIALS AND METHODS: We examined three types of treatment plans in 20 patients: (1) a six field plan, with a prescribed dose of 75.6 Gy; (2) the same six field plan to 72 Gy followed by a boost to 81 Gy; and (3) a five field plan with intensity modulated beams delivering 81 Gy. Treatment plans were designed using an initial CT data set (planning) and applied to three subsequent CT scans (treatment). The treatment CT contours were used to represent patient specific organ displacement; in addition, the dose distribution was convolved with a Gaussian distribution to model random setup error. Dose-volume histograms were calculated using an organ deformation model in which the movement between scans of individual points interior to the organs was tracked and the dose accumulated. The tumor control probability (TCP) for the prostate and proximal half of seminal vesicles (clinical target volume, CTV), normal tissue complication probability (NTCP) for the rectum and the percent volume of bladder wall receiving at least 75 Gy were calculated. RESULTS: The patient averaged increase in the planned TCP between plan types 2 and 1 and types 3 and 1 was 9.8% (range 4.9-12.5%) for both, whereas the corresponding increases in treatment TCP were 9.0% (1.3-16%) and 8.1% (-1.3-13.8%). In all patients, plans 2 and 3 (81 Gy) exhibited equal or higher treatment TCP than plan 1 (75.6 Gy). The maximum treatment NTCP for rectum never exceeded the planning constraint and percent volume of bladder wall receiving at least 75 Gy was similar in the planning and treatment scans for all three plans. CONCLUSION: For plans that deliver a uniform prescribed dose to the planning target volume (PTV) (plan 1), current margins are adequate. In plans that further escalate the dose to part of the PTV (plans 2 and 3), in a fraction of the cases the CTV dose increase is less than planned, yet in all cases the TCP values are higher relative to the uniform dose PTV (plan 1). Doses to critical organs remain within the planning criteria. PMID- 12742267 TI - The effect of waiting time on local control and survival in head and neck carcinoma patients treated with radiotherapy. AB - PURPOSE: To determine the influence of waiting time for radiotherapy on local control and survival in a cohort of patients with head and neck carcinoma of different locations and stages treated with radiotherapy. MATERIAL AND METHODS: Retrospective study of 797 patients with squamous cell carcinoma located in the oral cavity, pharynx or larynx, treated with radiotherapy, and with a minimum follow-up of 3 years. Local recurrence and survival were analyzed in function of the waiting time, defined as the interval between date of histologic diagnosis and date of radiotherapy. A univariate and multivariate analysis was carried out. RESULTS: Median waiting time to radiotherapy was 44 days (25 and 75% quartiles: 33 and 60 days). There were significant differences in the waiting time period in relation to the primary location and the local extension of the tumor. Both univariate and multivariate analysis showed that waiting time had no significant impact either on local control or survival. CONCLUSION: Within the range of the waiting time observed in our study, delay in the initiation of radiotherapy did not affect local control or survival in patients with head and neck carcinoma. PMID- 12742268 TI - Prevention of radiation induced xerostomia by surgical transfer of submandibular salivary gland into the submental space. AB - BACKGROUND AND PURPOSE: Xerostomia is a significant morbidity of radiation treatment in the management of head and neck cancers. We hypothesized that the surgical transfer of one submandibular salivary gland to the submental space, where it can be shielded from radiation treatment (XRT), would prevent xerostomia. MATERIALS AND METHODS: We conducted a prospective Phase II clinical trial and the patients were followed clinically with salivary flow studies and the University of Washington Quality of Life questionnaire. RESULTS: We report the results on 76 evaluable patients. The salivary gland transfer was done in 60 patients. Nine patients (of 60) did not have postoperative XRT and in eight patients (of 60) the transferred gland was not shielded from XRT due to proximity of disease. The median follow up is 14 months. Of the 43 patients with the salivary gland transfer and post-operative XRT with protection of the transferred gland, 81% have none or minimal xerostomia, and 19% developed moderate to severe xerostomia. Three patients (6.9%) developed local recurrence, five patients (11.6%) developed distant metastases and five patients (11.6%) have died. There were no complications attributed to the surgical procedure. CONCLUSION: Surgical transfer of a submandibular salivary gland to the submental space preserves its function and prevents the development of radiation induced xerostomia. PMID- 12742269 TI - Optimisation of conformal radiation therapy by intensity modulation: cancer of the larynx and salivary gland function. AB - PURPOSE: Prevention of damage to critical normal tissues is of paramount importance for the quality of life of patients irradiated for cancers in the head and neck. The purpose of this paper was to evaluate the parotid gland sparing 3D conformal radiation therapy technique (3DCRT) in a prospective study in node negative cancer of the larynx. MATERIALS AND METHODS: Twenty-six patients with node negative squamous cell cancer of the larynx were irradiated by a 3DCRT technique (class solution) to both sides of the neck (elective dose 46 Gy to levels II, III and IV) and primary tumour (70 Gy). Dose distributions of the major salivary glands were correlated with objective (stimulated whole saliva flow, WS) and subjective (questionnaire; visual analogue scale, VAS) salivary gland function. Apart from the clinically used 3DCRT technique, in order to optimise 3DCRT dose distributions, intensity modulated (IMRT) treatment plans were generated for the same patient population. Dose-volume histograms of 3DCRT and IMRT treatment plans were analysed and compared. RESULTS: For the 26 patients irradiated with the 3DCRT class solution technique: VAS scores and questionnaires reached their nadir 3 months post-radiotherapy; WS reached its nadir 6 months post-radiotherapy. WS flow rates improved significantly, but never normalised; 2 years post-treatment WS measurements were 48% of the pre-treatment values. VAS scores deteriorated during ERT from 0 pre-treatment to 6.1 immediately post treatment. Compared to pre-treatment, questionnaires were answered affirmative by increasing numbers of patients. For all patients, IMRT treatment plans resulted in a significant reduction of the dose delivered to the parotid glands compared to the 3DCRT-treatment technique. CONCLUSIONS: The class solution for the 3DCRT salivary gland sparing technique is inadequate for fully preserving salivary gland function, given the dose distributions (DVHs) as well as the subjective- and objective salivary gland function assessments. The results can be optimised in the future, that is a further reduction of xerostomia can be achieved, by using IMRT techniques focused at sparing major and minor salivary glands. PMID- 12742270 TI - Systematic set-up errors for IMRT in the head and neck region: effect on dose distribution. AB - BACKGROUND AND PURPOSE: There is a general concern about intensity modulated radiation therapy (IMRT) treatments being more sensitive to patient positioning than conventional treatments. The aim of this study was to evaluate the International Commission on radiation units and measurements (ICRU) method for taking systematic set-up errors into account for IMRT treatments and to compare the effects on the dose distribution with the effects of conventional treatments. MATERIAL AND METHODS: A planning margin to account for set-up errors was added to the clinical target volumes and to the spinal cords, for three head and neck patients, according to the ICRU. No margin was added to organs at risk with mainly parallel structure if they were situated adjacent to the target volume, for example, the parotid glands. The effects of set-up errors in six IMRT plans and three conventional plans were simulated in the planning system and analysed with physical dose parameters. RESULTS AND CONCLUSIONS: In general, the ICRU method of taking set-up errors into account works satisfactorily for IMRT treatments as well as for conventional treatments with no difference between the treatment techniques. The sensitivity to set-up errors regarding the target volume is dependent on the quality of the treatment plan, i.e. the part of the target covered with a dose >95 and <105% and the effect in the critical organs is dependent on the sharpness of the dose gradients outside the critical organ. However, the method makes it difficult to include organs at risk with mainly parallel structure if they are situated adjacent to the target volume. PMID- 12742271 TI - Stereotactic intensity modulated radiation therapy and inverse treatment planning for tumors of the head and neck region: clinical implementation of the step and shoot approach and first clinical results. AB - PURPOSE/OBJECTIVE: The aim of this analysis is to evaluate the feasibility of inverse treatment planning and intensity modulated radiation therapy (IMRT) for head and neck cancer in daily clinical routine. A step and shoot IMRT approach was developed which allows the treatment of large target volumes without the need to use a split beam technique. By using the IMRT approach better protection of different organs at risk in the head and neck region may be achieved and an escalation of the dose in the tumor should be possible. We evaluated the feasibility of the treatment technique and the patient tolerance to the treatment. First clinical results are reported. MATERIALS AND METHODS: Between 1999 and 2002, 48 patients with a carcinoma of the head and neck region were treated with curative intention. All patients were treated in a patient-specific Scotch-Cast mask. Patients who required treatment of the lymph node levels I-VI, were additionally positioned by a vacuum pillow in order to immobilize the upper part of the thorax. For inverse treatment planning, the software module KonRad was used which was integrated into the VIRTUOS planning system. Each treatment plan was verified using quantitative film dosimetry in a head and neck phantom. The step and shoot IMRT technique with a multileaf collimator integrated in a Primus (Siemens) accelerator was used for treatment. For all target volumes the whole target including the lymph nodes were covered completely by the IMRT treatment. RESULTS: The mean total dose for the target volumes of macroscopic disease ranged between 63.0 and 64.1 Gy. The mean total dose of microscopic disease ranged between 55.2 and 60.1 Gy. The mean percentage of planning target volume receiving <90% of the prescribed dose ranged between 3.0 and 11.5%. For the treatment, the median number of beams was seven (range: five to nine). The time to deliver the treatment ranged between 9 and 18 min. The results of the verification revealed a mean deviation between measured and calculated absolute doses for the 48 patients of 0.1+/-1.4%. Including the phantom verification the IMRT treatment of the patients could be started approximately after five working days. The treatment was well tolerated by all patients. The 2-year actuarial overall survival was 92% and the 2-year actuarial local control rate was 93%. According to the Radiation Therapy Oncology Group (RTOG), no higher acute toxicity than Grade 3 was seen. Observation of the late effects revealed only one transient Grade 4 toxicity of the bone and only four patients had a xerostomia higher than Grade 1. CONCLUSION: The use of an inversely-planned and intensity modulated step and shoot approach is feasible in clinical routine for head and neck tumors. Treatment could be applied as planned and no increased toxicity was found. Compared to other IMRT approaches for the head and neck region the used technique allows the treatment of the primary tumor and the lymph nodes level I VI with only one intensity modulated treatment volume. The presented technique avoids to match conventional radiotherapy fields and IMRT fields, and therefore, reduce the risk of overdosage or underdosage at the matching line. Compared to conventional treatment techniques IMRT shows advantages in tumor dose and dose at the organs at risk. PMID- 12742272 TI - Follow-up of head and neck cancer patients post-radiotherapy. AB - Diverging opinions exist regarding follow-up studies post-radiotherapy for head and neck cancer. This report describes the efficacy of follow-up physical examinations, thyroid function tests and screening chest X-rays in post radiotherapy patients in a practice analysis schema. This analysis suggests that physical examination and thyroid function testing remain valid parts of routine follow-up for head and neck cancer patients; chest X-rays appear less vital unless the patient's clinical situation warrants aggressive therapy of a second primary lung cancer. PMID- 12742273 TI - High dose rate brachytherapy for the palliation of malignant dysphagia. AB - BACKGROUND AND PURPOSE: High dose rate (HDR) brachytherapy is a commonly used palliative treatment for esophageal carcinoma. We evaluated the outcome of HDR brachytherapy in patients with malignant dysphagia. MATERIAL AND METHODS: A retrospective analysis over a 10-year period was performed of 149 patients treated with HDR brachytherapy, administered in one or two sessions, at a median dose of 15Gy. Patients were evaluated for functional outcome, complications, recurrent dysphagia, and survival. RESULTS: At 6 weeks after HDR brachytherapy, dysphagia scores had improved from a median of 3 to 2 (n=104; P<0.001), however, dysphagia had not improved in 51 (49%) patients. Procedure-related complications occurred in seven (5%) patients. Late complications, including fistula formation or bleeding, occurred in 11 (7%) patients. Twelve (8%) patients experienced minor retrosternal pain. Median survival of the patients was 160 days with a 1-year survival rate of 15%. Procedure-related mortality was 2%. At follow-up, 55 (37%) patients experienced recurrent dysphagia. In 34 (23%) patients a metal stent was placed to relieve persistent or recurrent dysphagia. CONCLUSION: HDR brachytherapy is a moderately effective treatment for the palliation of malignant dysphagia. The incidence of early major complications is low, however, persistent and recurrent dysphagia occur frequently, and require often additional treatment. PMID- 12742274 TI - Radiotherapy of Hodgkin's disease in early pregnancy: embryo dose measurements. AB - BACKGROUND AND PURPOSE: Limited information exists on the possibility of pregnant women undergoing radiotherapy for Hodgkin's disease in early pregnancy. The purpose of this study was to measure embryo dose resulting from treatment of supra-diaphragmatic Hodgkin's disease at the first trimester of gestation. MATERIALS AND METHODS: A humanoid phantom was used to simulate pregnancy at the first trimester of gestation. Embryo dose, was measured using three different field sizes that may be applied for local field irradiation in each of the regions of neck, axilla, neck-mediastinum and for mantle treatment. A shielding device consisting of 5 cm of lead was used to reduce the embryo dose. Dose measurements were carried out using thermoluminescent dosimeters. Phantom exposures were made with a 6 MV photon beam. RESULTS: Local field irradiation in the regions of neck or axilla always resulted in embryo doses below 10 cGy. For local field irradiation in the region of neck-mediastinum and for mantle treatment, the radiation dose to a shielded embryo was 2.8-18.6 and 4.2-24.5 cGy depending upon the distance from the field isocenter and the field size used, respectively. The corresponding dose for an unshielded embryo exceeded 10 cGy. All the above embryo doses were obtained for a tumor dose of 40 Gy. CONCLUSIONS: Local field irradiation in the regions of neck or axilla may be safely performed even without uterus shielding. For local field irradiation in the region of neck mediastinum and for mantle radiotherapy, the extent of the irradiated area, the distance separating the embryo from the field isocenter and the tumor dose are the factors, determining whether the radiation dose to a shielded embryo may possibly be reduced below 10 cGy. PMID- 12742275 TI - Chromosomal fragility syndrome and family history of radiosensitivity as indicators for radiotherapy dose modification. AB - Beside a few known radiosensitive syndromes, a patient's reaction to radiotherapy is difficult to predict. In this report we describe the management of a pediatric cancer patient presented with a family history of radiosensitivity and cancer proneness. Laboratory investigations revealed a chromosomal fragility syndrome and an increased cellular radiosensitivity in vitro. AT gene sequencing revealed no mutations. The patient was treated with reduced radiation doses to avoid the presumed increased risks of toxicity to normal tissues. The patient tolerated well the treatment with no significant acute or late radiation sequelae. Five years later, the patient remains both disease and complications free. While an accurate laboratory test for radiosensitivity is still lacking, assessments of chromosomal fragility, cell survival and clinical medicine will continue to be useful for a small number of patients. PMID- 12742276 TI - Infrastructure of radiotherapy in the Netherlands: evaluation of prognoses and introduction of a new model for determining the needs. AB - BACKGROUND AND PURPOSE: In the Netherlands, the radiotherapy infrastructure is regulated by a Governmental license system. This requires timely and realistic prognostication of the needs for radiotherapy. In the present study, the latest prognoses (1993) and the realized changes in infrastructure are evaluated and a new prognosis for the period until 2010, which has been calculated using a new model, is presented. MATERIALS AND METHODS: Data on cancer incidence and use of radiotherapy were obtained from various published national reports and from a survey of all radiotherapy departments. RESULTS: The cancer incidence over the period 1993-1997 was about 10% higher than predicted. In 1996 and 1997, the percentage of new cancer patients treated with radiotherapy was 45.6 and 48.2%, respectively. The absolute number of newly irradiated patients was about 10% higher than foreseen in the prognosis. The needs for radiotherapy infrastructure not only depend on epidemiological data and changes in indications for radiotherapy, but also on changes in types of treatment with different workloads. A new model, which uses four categories for teletherapy and four categories for brachytherapy is described and a new prognosis for the required number of linear accelerators and staff up to the year 2010 is presented. CONCLUSION: The original prognosis on cancer incidence and radiotherapy patients underestimated the actual figures considerably. The new prognosis, based on a model, which not only accounts for an increase in number of patients, but also for changes in treatment techniques, is expected to more accurately predict and acquire the required radiotherapy capacity. PMID- 12742277 TI - Obstructive sleep apnea and cardiovascular disease. AB - Obstructive sleep apnea (OSA) is a common disorder associated with an increased risk of cardiovascular disease and stroke. As it is strongly associated with known cardiovascular risk factors, including obesity, insulin resistance, and dyslipidemia, OSA is an independent risk factor for hypertension and has also been implicated in the pathogenesis of congestive cardiac failure, pulmonary hypertension, arrhythmias, and atherosclerosis. Obesity is strongly linked to an increased risk of OSA, and weight loss can reduce the severity of OSA. The current standard treatment for OSA-nasal continuous positive airway pressure (CPAP)-eliminates apnea and the ensuing acute hemodynamic changes during sleep. Long-term CPAP treatment studies have shown a reduction in nocturnal cardiac ischemic episodes and improvements in daytime blood pressure levels and left ventricular function. Despite the availability of effective therapy, OSA remains an underdiagnosed and undertreated condition. A lack of physician awareness is one of the primary reasons for this deficit in diagnosis and treatment. PMID- 12742278 TI - Carvedilol increases two-year survivalin dialysis patients with dilated cardiomyopathy: a prospective, placebo-controlled trial. AB - OBJECTIVES: We sought to evaluate the effects of carvedilol on mortality and morbidity in dialysis patients with dilated cardiomyopathy. BACKGROUND: Several lines of evidence support the concept that therapy with beta-blocking agents reduces morbidity and mortality in patients with congestive heart failure (HF), but the demonstration of such a survival benefit in dialysis patients with dilated cardiomyopathy is still lacking. METHODS: A total of 114 dialysis patients with dilated cardiomyopathy were randomized to receive either carvedilol or placebo in addition to standard therapy. A first analysis was performed at one year and was followed by an additional follow-up period of 12 months. RESULTS: Two-year echocardiographic data revealed a significant attenuation of pathologic remodeling, with smaller cavity diameters and higher ejection fractions in the active treatment group than in the placebo group. At two years, 51.7% of the patients died in the carvedilol group, compared with 73.2% in the placebo group (p < 0.01). Furthermore, there were significantly fewer cardiovascular deaths (29.3%) and hospital admissions (34.5%) among patients receiving carvedilol than among those receiving a placebo (67.9% and 58.9%, respectively; p < 0.00001). The exploratory analyses revealed that fatal myocardial infarctions, fatal strokes, and hospital admissions for worsening HF were lower in the carvedilol group than in the placebo group. A reduction in sudden deaths and pump-failure deaths was also observed, though it did not reach statistical significance. CONCLUSIONS: Carvedilol reduced morbidity and mortality in dialysis patients with dilated cardiomyopathy. These data suggest the use of carvedilol in all dialysis patients with chronic HF. PMID- 12742279 TI - Ximelagatran versus warfarin for stroke prevention in patients with nonvalvular atrial fibrillation. SPORTIF II: a dose-guiding, tolerability, and safety study. AB - OBJECTIVES: We sought to compare the tolerability and safety of three fixed doses of ximelagatran versus warfarin in patients with nonvalvular atrial fibrillation (NVAF). BACKGROUND: Anticoagulants such as warfarin lower the risk of stroke in patients with NVAF. Ximelagatran is a novel, oral direct thrombin inhibitor with predictable pharmacokinetics and no known food or pharmacokinetic drug interactions. METHODS: This was a 12-week, randomized, parallel-group, dose guiding study of NVAF patients with at least one additional risk factor for stroke. The primary end point was the number of thromboembolic events and bleedings. Three groups received ximelagatran (n = 187) at 20, 40, or 60 mg twice daily, given in a double-blind fashion, without routine coagulation monitoring. In a fourth group, warfarin (n = 67) was managed and monitored according to normal routines, aiming for an International Normalized Ratio of 2.0 to 3.0. RESULTS: A total of 254 patients received study drug. One ischemic stroke (nonfatal) and one transient ischemic attack (TIA) occurred in the ximelagatran group. Two TIAs occurred in the warfarin group. No major bleeds were observed in the ximelagatran group. One major bleed occurred in a warfarin-treated patient. The number of minor and multiple minor bleeds was low, but there was a slight increase by ximelagatran dose. The 60-mg dose resulted in the same number of bleeding events as that with warfarin. S-alanine aminotransferase was increased in eight patients (4.3%) taking ximelagatran, but normalized with continuous treatment or cessation of the drug. CONCLUSIONS: Fixed oral doses of ximelagatran up to 60 mg twice daily were well tolerated, without the need for dose adjustment or coagulation monitoring. PMID- 12742281 TI - The neurohormonal paradigm: have we gone too far? PMID- 12742280 TI - Tezosentan in patients with acute heart failure and acute coronary syndromes: results of the Randomized Intravenous TeZosentan Study (RITZ-4). AB - OBJECTIVES: We sought to determine the effect of tezosentan in patients with acute decompensated heart failure (HF) associated with acute coronary syndrome (ACS). BACKGROUND: Tezosentan is a dual endothelin receptor antagonist that has been shown to improve cardiac output, decrease pulmonary capillary wedge pressure, and reduce pulmonary and systemic vascular resistance in initial clinical studies in acute decompensated HF. METHODS: The Randomized Intravenous TeZosentan (RITZ)-4 study was a multicenter, randomized, double-blinded, placebo controlled study of tezosentan in patients with acute decompensated HF associated with ACS. A total of 193 patients were randomized to receive tezosentan (25 mg/h for 1 h, then 50 mg/h for 23 to 47 h) or placebo. Patients with evidence of acute decompensated HF and ACS were eligible to participate. The primary end point was the composite of death, worsening HF, recurrent ischemia, and recurrent or new myocardial infarction within 72 h. RESULTS: No significant differences were observed between placebo and 50 mg/h tezosentan in the composite primary end point: 24.2% (95% confidence interval [CI] 16.0% to 34.1%) and 28.9% (95% CI 20.1% to 39.0%), respectively (p = 0.5152). Symptomatic hypotension was more frequent in the treatment group. CONCLUSIONS: At the doses studied, tezosentan did not result in a significant improvement in the composite primary clinical end point in the RITZ-4 trial. Tezosentan did not demonstrate pro-ischemic effects in this population. Symptomatic hypotension may have resulted in an increased number of adverse events in the treatment group. Further studies with lower tezosentan doses are warranted. PMID- 12742282 TI - Hydroxymethylglutaryl coenzyme a reductase inhibitors down-regulate chemokines and chemokine receptors in patients with coronary artery disease. AB - OBJECTIVES: We sought to investigate whether the activation of the chemokine network observed in patients with coronary artery disease (CAD) could be modified by treatment with 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins). BACKGROUND: Chemokines and chemokine receptors are important mediators in atherogenesis, and we hypothesized that the statins could affect the chemokine network in CAD. METHODS: Thirty CAD patients without previous statin therapy were randomized to receive atorvastatin (80 mg/day, n = 15) or simvastatin (20 mg/day, n = 15). Peripheral blood mononuclear cells (PBMCs) and plasma were obtained at baseline and after six months of statin therapy. Messenger ribonucleic acid (mRNA) expression of chemokines and chemokine receptors in PBMCs was analyzed by ribonuclease protection assay and real-time reverse-transcription polymerase chain reaction. Chemokines were also examined in the supernatants from unstimulated and lipopolysaccharide-stimulated PBMCs (and in plasma). RESULTS: Our main findings were: 1) gene expression of several chemokines (i.e., macrophage inflammatory protein [MIP]-1alpha, MIP-1beta, and interleukin [IL]-8) and chemokine receptors (i.e., CC chemokine receptor [CCR]1, CCR2, CCR4, and CCR5) was markedly increased among CAD patients compared with healthy control subjects; 2) treatment with atorvastatin and simvastatin markedly reduced the mRNA levels of some of these chemokines (i.e., MIP-1alpha, MIP-1beta, IL-8) and receptors (i.e., CCR1 and CCR2), with the most pronounced effect in the atorvastatin group; and 3) statin therapy reduced the spontaneous release of IL-8 and MIP-1alpha from PBMCs in CAD patients. CONCLUSIONS: This study demonstrates a down-regulatory effect of statins on the chemokine network in CAD patients, possibly contributing to the beneficial effects of statins in this disorder. PMID- 12742283 TI - C-reactive protein, clinical presentation, and ischemic activity in patients with chest pain and normal coronary angiograms. AB - OBJECTIVES: We sought to investigate the relationship among C-reactive protein (hs-CRP), clinical characteristics, exercise stress test responses, and ST segment changes during daily life in patients with typical chest pain and normal coronary angiograms (CPNCA). BACKGROUND: Patients with CPNCA have coronary microvascular endothelial dysfunction and myocardial ischemia. Elevated hs-CRP levels have been related to atherogenesis and endothelial dysfunction. The relationship between hs-CRP and disease activity has not been previously investigated in CPNCA patients. METHODS: We studied 137 consecutive CPNCA patients (mean age, 57 +/- 9; 33 men). All completed standardized angina questionnaires, underwent exercise stress testing, 24-h ambulatory electrocardiogram (ECG) monitoring (Holter), and hs-CRP measurements at study entry. RESULTS: C-reactive protein levels (mg/l) were higher in patients with frequent (2.9 +/- 3.3) and prolonged (3.9 +/- 4.1) chest pain episodes, and in those with ST-segment depression on exercise testing (2.6 +/- 2.8) and Holter monitoring (3.4 +/- 3.1) compared with patients with occasional (1.3 +/- 1.2; p = 0.002) or shorter chest pain (1.5 +/- 1.3; p < 0.001) episodes, negative exercise stress testing (1.1 +/- 1.1; p < 0.001), and no ST-segment shifts on Holter monitoring (0.9 +/- 0.7; p < 0.001). Moreover, we found a correlation between hs CRP concentration and number of ischemic episodes during Holter monitoring (r = 0.65; p < 0.001) and with the magnitude of ST-segment depression on exercise testing (r = -0.43; p < 0.001). The hs-CRP was the only independent variable (multivariate logistic regression) capable of predicting positive findings on Holter monitoring (odds ratio [OR], 3.8; confidence interval [CI], 2.3 to 6.2) and exercise testing (OR, 1.7; CI, 1.2 to 2.2). CONCLUSIONS: The hs-CRP correlates with symptoms and ECG markers of myocardial ischemia in CPNCA patients. Whether hs-CRP is related to the pathogenesis of angina in these patients deserves further investigation. PMID- 12742284 TI - Preventing myocardial infarction in the young adult in the first place: how do the National Cholesterol Education Panel III guidelines perform? AB - OBJECTIVES: The purpose of this study was to investigate the utility of the new National Cholesterol Education Program (NCEP) III guidelines in a group of young adults. BACKGROUND: These guidelines have been hailed as an improvement in their potential to identify individuals at risk for coronary heart disease (CHD) complications. Compared with the NCEP II, the new guidelines will increase the number of patients who qualify for medical management. However, the effectiveness of these guidelines to identify young adults at risk for a cardiac event is yet to be studied. METHODS: A retrospective review of clinical data from young adults (age or=1:16) were detected in 287 subjects (74.5%): low titers (1:16 to 1:32) in 58 (15.1%), intermediate titers (1:64 to 1:128) in 144 (37.4%), and high titers (>or=1:256) in 85 subjects (22.1%). Antibody titers were not associated with the presence of aortic plaques after adjustment for age, gender, and smoking status (p = 0.64). Compared with titers <1:16, the adjusted odds ratios for aortic plaques were 1.46 (95% confidence interval [CI] 0.63 to 3.42) for low titers, 1.32 (95% CI 0.68 to 2.55) for intermediate titers, and 0.94 (95% CI 0.42 to 2.07) for high titers. Among the subgroup with plaques, antibody titers were not associated with the presence of plaques >or=4 mm thick (p = 0.99), plaques >or=6 mm (p = 0.49), or mobile debris (p = 0.71), after adjustment for age and smoking. CONCLUSIONS: Chlamydia pneumoniae IgG antibody titers are not associated with the presence or severity of aortic atherosclerosis in the general population. These observations do not support a role for C pneumoniae infection in the initiation or progression of atherosclerosis. PMID- 12742287 TI - Coronary stent implantation is superior to balloon angioplasty for chronic coronary occlusions: six-year clinical follow-up of the GISSOC trial. AB - OBJECTIVES: We investigated whether the benefits of stent implantation over balloon percutaneous transluminal coronary angioplasty (PTCA) for treatment of chronic total coronary occlusions (CTO) are maintained in the long term. BACKGROUND: Several randomized trials have shown that in CTO, stent implantation confers clinical and angiographic mid-term outcomes superior to those observed after PTCA. However, limited information on the long-term results of either technique is available. METHODS: Six-year clinical follow-up of patients enrolled in the Gruppo Italiano di Studio sullo Stent nelle Occlusioni Coronariche (GISSOC) trial was performed by direct visit or telephone interview. Major adverse cardiac events (MACE), defined as cardiac death, myocardial infarction, target lesion revascularization (TLR), and anginal status, were recorded. RESULTS: Freedom from MACE at six years was 76.1% in the stent group, compared with 60.4% in the PTCA group (p = 0.0555). This difference was due mainly to TLR free survival rates (85.1% vs. 65.5% for the stent and PTCA groups, respectively; p = 0.0165). Eleven patients underwent TLR after the nine-month follow-up visit (stent group: n = 5; PTCA group: n = 6); however, in most cases, restenosis of the study occlusion was evident at nine-month angiography. CONCLUSIONS: This study represents the longest reported clinical follow-up of patients after percutaneous recanalization of CTO and demonstrates that the superiority of stent implantation over balloon PTCA is maintained in the long term. Stent and PTCA results appear to remain stable after nine-month angiographic follow-up. Stent implantation in CTO that can be recanalized percutaneously is therefore a valuable long-term therapeutic option. PMID- 12742288 TI - Stenting chronic coronary artery occlusions. One step closer? PMID- 12742289 TI - Calcium antagonists reduce cardiovascular complications after cardiac surgery: a meta-analysis. AB - OBJECTIVES: We sought to determine the efficacy of calcium antagonists (CAs) in reducing death, myocardial infarction (MI), ischemia, and supraventricular tachyarrhythmia (SVT) after cardiac surgery. BACKGROUND: Calcium antagonists may reduce complications after cardiac surgery-namely, death, MI, and renal failure. However, they are underused, possibly due to the results from previous observational studies. METHODS: Both MEDLINE (1966 to December 2001) and EMBASE (1980 to December 2001) were searched, with supplementation by reference list searches. No language restrictions were applied. Included studies were randomized, controlled trials (RCTs) evaluating preoperative, intraoperative, or postoperative (first 48 h) CA use (intravenous or oral) during aortocoronary bypass or valve surgery. Studies were excluded if they exclusively recruited transplant recipients, individuals <18 years old, or patients with pre-existing SVT. Two reviewers independently evaluated study quality by using the Jadad score; a minimal score of 1/5 was required. Forty-one studies, encompassing 3,327 patients, were included. No studies assessed treatment exclusively with short acting oral nifedipine. Treatment effects were calculated using the random effects model. Heterogeneity was assessed using the Q test. RESULTS: Calcium antagonists significantly reduced MI (odds ratio [OR] 0.58, 95% confidence interval [CI] 0.37 to 0.91; p = 0.02) and ischemia (OR 0.53, 95% CI 0.39 to 0.72; p < 0.001). Non-dihydropyridines significantly reduced SVT (OR 0.62, 95% CI 0.41 to 0.93; p = 0.02). Calcium antagonists were associated with trends toward decreased mortality during aortocoronary bypass (OR 0.66, 95% CI 0.26 to 1.70, p = 0.4). CONCLUSIONS: Use of CAs during cardiac surgery significantly reduced rates of MI, ischemia, and SVT. Further study using large RCTs is justified. PMID- 12742290 TI - Anti-ischemic properties of calcium-channel blockers: lessons from cardiac surgery. PMID- 12742291 TI - Outcomes in heart failure patients with preserved ejection fraction: mortality, readmission, and functional decline. AB - OBJECTIVES: We evaluated the six-month clinical trajectory of patients hospitalized for heart failure (HF) with preserved ejection fraction (EF), as the natural history of this condition has not been well established. We compared mortality, hospital readmission, and changes in functional status in patients with preserved versus depressed EF. BACKGROUND: Although the poor prognosis of HF with depressed EF has been extensively documented, there are only limited and conflicting data concerning clinical outcomes for patients with preserved EF. METHODS: We prospectively evaluated 413 patients hospitalized for HF to determine whether EF >or=40% was an independent predictor of mortality, readmission, and the combined outcome of functional decline or death. RESULTS: After six months, 13% of patients with preserved EF died, compared with 21% of patients with depressed EF (p = 0.02). However, the rates of functional decline were similar among those with preserved and depressed EF (30% vs. 23%, respectively; p = 0.14). After adjusting for demographic and clinical covariates, preserved EF was associated with a lower risk of death (hazard ratio [HR] 0.49, 95% confidence interval [CI] 0.26 to 0.90; p = 0.02), but there was no difference in the risk of readmission (HR 1.01, 95% CI 0.72 to 1.43; p = 0.96) or the odds of functional decline or death (OR 1.01, 95% CI 0.59 to 1.72; p = 0.97). CONCLUSIONS: Heart failure with preserved EF confers a considerable burden on patients, with the risk of readmission, disability, and symptoms subsequent to hospital discharge, comparable to that of HF patients with depressed EF. PMID- 12742292 TI - Heart failure with preserved ejection fraction: is this diastolic heart failure? PMID- 12742293 TI - Cardiac resynchronization therapy homogenizes myocardial glucose metabolism and perfusion in dilated cardiomyopathy and left bundle branch block. AB - OBJECTIVES: We investigated whether cardiac resynchronization therapy (CRT) affects myocardial glucose metabolism and perfusion in dilated cardiomyopathy (DCM) and left bundle branch block (LBBB). BACKGROUND: Patients with DCM and LBBB present with asynchronous left ventricular (LV) activation, leading to reduced septal glucose metabolism. Cardiac resynchronization therapy recoordinates LV activation, but its effects on myocardial glucose metabolism and perfusion remain unknown. METHODS: In 15 patients (10 females; 61 +/- 13 years) with DCM and LBBB (QRS width 165 +/- 15 ms), gated (18)F-fluorodeoxyglucose (FDG) positron emission tomography (PET) and technetium-99m ((99m)Tc)-sestamibi single-photon emission computed tomography were performed before and after two weeks of CRT. Uptake of FDG and (99m)Tc-sestamibi was determined in four LV wall areas. Ejection fraction and volumes were calculated from gated PET. RESULTS: Baseline FDG uptake was heterogeneous (p < 0.0001), with lowest uptake in the septal region (56 +/- 12%) and highest uptake in the lateral region (89 +/- 6%). During CRT, septal and anterior increases (p < 0.01) and lateral decreases (p < 0.01) resulted in homogeneously distributed glucose metabolism. Baseline heterogeneity (p < 0.0001) in (99m)Tc-sestamibi uptake was modest (lowest septal 65 +/- 10%; maximum lateral 84 +/- 5%) and also reduced with CRT, although some heterogeneity (p < 0.05) remained. The septal-to-lateral ratio increased with CRT for FDG (0.62 +/- 0.12 to 0.91 +/- 0.26, p < 0.001) and (99m)Tc-sestamibi uptake (0.77 +/- 0.13 to 0.85 +/- 0.16, p < 0.01). The LV end-diastolic and end-systolic volumes decreased from 293 +/- 160 to 272 +/- 158 ml (p < 0.05) and from 244 +/- 164 to 220 +/- 160 ml (p < 0.01), respectively. Ejection fraction increased from 22 +/- 12% to 25 +/- 13% (p < 0.01). CONCLUSIONS: Glucose metabolism is reduced more than perfusion in the septal compared with LV lateral wall in patients with DCM and LBBB. Cardiac resynchronization therapy restores homogeneous myocardial glucose metabolism with less influence on perfusion. PMID- 12742294 TI - Efficacy of angiotensin-converting enzyme inhibitors and beta-blockers in the management of left ventricular systolic dysfunction according to race, gender, and diabetic status: a meta-analysis of major clinical trials. AB - OBJECTIVES: This study sought to assess the effect of angiotensin-converting enzyme (ACE) inhibitors and beta-blockers on all-cause mortality in patients with left ventricular (LV) systolic dysfunction according to gender, race, and the presence of diabetes. BACKGROUND: Major randomized clinical trials have established that ACE inhibitors and beta-blockers have life-saving benefits in patients with LV systolic dysfunction. Most patients enrolled in these trials were Caucasian men. Whether an equal effect is achieved in women, non-Caucasians, and patients with major comorbidities has not been established. METHODS: The authors performed a meta-analysis of published and individual patient data from the 12 largest randomized clinical trials of ACE inhibitors and beta-blockers to produce random effects estimates of mortality for subgroups. RESULTS: Data support beneficial reductions in all-cause mortality for the use of beta-blockers in men and women, the use of ACE inhibitors and some beta-blockers in black and white patients, and the use of ACE inhibitors and beta-blockers in patients with or without diabetes. Women with symptomatic LV systolic dysfunction probably benefit from ACE inhibitors, but women with asymptomatic LV systolic dysfunction may not have reduced mortality when treated with ACE inhibitors (pooled relative risk = 0.96; 95% confidence interval: 0.75 to 1.22). The pooled estimate of three beta-blocker studies supports a beneficial effect in black patients with heart failure, but one study assessing bucindolol reported a nonsignificant increase in mortality. CONCLUSIONS: Angiotensin-converting enzyme inhibitors and beta blockers provide life-saving benefits in most of the subpopulations assessed. Women with asymptomatic LV systolic dysfunction may not achieve a mortality benefit when treated with ACE inhibitors. PMID- 12742295 TI - Effect of a change in gender on coronary arterial size: a longitudinal intravascular ultrasound study in transplanted hearts. AB - OBJECTIVES: We sought to document whether a physiologic change in gender has any effect on coronary arterial size. BACKGROUND: The coronary arteries are smaller in women, even after correction for body surface area (BSA). These differences may contribute to adverse clinical outcomes after coronary artery bypass graft surgery and myocardial infarction in women. In male and female transsexuals, pharmacologic doses of estrogens and androgens significantly influence vascular diameter. Thus, gender differences in the coronary vasculature may be a reflection of the hormonal environment. METHODS: In 86 patients who had undergone orthotopic heart transplantation, serial intravascular ultrasound studies of the proximal left anterior descending coronary artery (LAD) were analyzed. Changes in vessel area (VA) over the first or second post-transplant year were recorded, and comparisons were made between donor hearts that were transplanted in a patient of the same gender and those that were transplanted in a patient of the opposite gender. RESULTS: Vessel area of the proximal LAD increased over time in all patient groups. In hearts transplanted within the same gender and in male donor hearts transplanted to female recipients, the change was small and not significant. However, in hearts transplanted from female donors to male recipients, there was a substantial and highly significant increase in LAD VA (median 16.13 to 17.88 mm(2); p = 0.01). This increase was not explained by confounding due to changes in BSA or left ventricular wall thickness. CONCLUSIONS: This pattern of arterial remodeling early after heart transplantation supports a link between host gender and coronary arterial size. PMID- 12742296 TI - The metabolic syndrome, diabetes, and subclinical atherosclerosis assessed by coronary calcium. AB - OBJECTIVES: We compared the prevalence and extent of coronary artery calcium (CAC) among persons with the metabolic syndrome (MetS), diabetes, and neither condition. BACKGROUND: The prevalence and extent of CAC has not been compared among those with MetS, diabetes, or neither condition. METHODS: Of 1,823 persons (36% female) age 20 to 79 years who had screening for CAC by computed tomography, 279 had MetS, 150 had diabetes, and the remainder (n = 1,394) had neither condition. Metabolic syndrome was defined with >or=3 of the following: body mass index >or=30 kg/m(2); high-density lipoprotein cholesterol <40 mg/dl if male or <50 mg/dl if female; triglycerides >or=150 mg/dl; blood pressure >or=130/85 mm Hg or on treatment; or fasting glucose 110 to 125 mg/dl. The prevalence and odds of any and significant (>or=75th percentile) CAC among these groups and by number of MetS risk factors were determined. RESULTS: Those with neither MetS nor diabetes, MetS, or diabetes had a prevalence of CAC of 53.5%, 58.8%, and 75.3% (p < 0.001), respectively, among men and 37.6%, 50.8%, and 52.6% (p < 0.001), respectively, among women. Coronary artery calcium increased by the number (0 to 5) of MetS risk factors (from 34.0% to 58.3%) (p < 0.001). Forty-one percent of subjects with MetS had either a >20% 10-year risk of CHD or CAC >or=75th percentile for age and gender. Risk factor-adjusted odds for the presence of CAC were 1.40 (95% confidence interval [CI] 1.05 to 1.87) among those with MetS and 1.67 (95% CI 1.12 to 2.50) among those with diabetes, versus those with neither condition. CONCLUSIONS: Those with MetS or diabetes have an increased likelihood of CAC compared with those having neither condition. PMID- 12742297 TI - Assessment of coronary flow reserve by coronary pressure measurement: comparison with flow- or velocity-derived coronary flow reserve. AB - OBJECTIVES: This study sought to assess the reliability of pressure-derived coronary flow reserve (CFR) compared with flow- or velocity-derived CFR. BACKGROUND: Coronary flow reserve has been reported to have important clinical implications for the evaluation and treatment of coronary artery disease. METHODS: Using a pressure guide wire, coronary pressure distal to the stenosis was measured at rest and during hyperemia in seven dogs with various degrees of stenosis and in 30 patients with angina (29 and 34 stenoses in total, respectively). Pressure at the tip of the guiding catheter was also recorded with a fluid-filled transducer system. Pressure-derived CFR was calculated by the square root of the pressure gradient across the stenosis (DeltaP) during hyperemia divided by DeltaP at rest, using a proprietary software system. At the same time, coronary flow was monitored proximal to the stenosis with a flow meter in the experimental dogs, and coronary flow velocity distal to the stenosis was assessed using a Doppler guide wire in patients with angina. Flow-derived (or velocity-derived) CFR was compared with pressure-derived CFR. RESULTS: Except for one stenosis that showed no DeltaP at rest, a significant correlation was obtained between pressure- and flow-derived CFR in the animal study (y = 1.05x - 0.03, r = 0.92, p = 0.0001). A significant correlation was also seen between pressure- and velocity-derived CFR in the human study, except in three stenoses with no resting DeltaP (y = 0.70x + 0.37, r = 0.85, p = 0.0001). CONCLUSIONS: Similar to flow (or velocity) measurement, CFR can be assessed by pressure measurement, except in stenoses with minor resting DeltaP. PMID- 12742298 TI - Toward clinical risk assessment in hypertrophic cardiomyopathy with gadolinium cardiovascular magnetic resonance. AB - OBJECTIVES: We sought to assess whether hyperenhancement by gadolinium cardiovascular magnetic resonance (CMR) occurs in hypertrophic cardiomyopathy (HCM) and correlates with the risk of heart failure and sudden death. BACKGROUND: The myocardial interstitium is abnormal in HCM at post-mortem. Focally increased interstitial myocardial space appears as hyperenhancement with gadolinium CMR. METHODS: In a blinded, prospective study, HCM patients were selected for the presence (n = 23) or absence (n = 30) of an increased clinical risk of sudden death and/or progressive adverse left ventricular (LV) remodeling. Gadolinium enhanced CMR was performed. RESULTS: Myocardial hyperenhancement was found in 42 patients (79%), affecting 10.9% (range 0% to 48%) of the LV mass. There was a greater extent of hyperenhancement in patients with progressive disease (28.5% vs. 8.7%, p < 0.001) and in patients with two or more risk factors for sudden death (15.7% vs. 8.6%, p = 0.02). Improved discrimination was seen in patients >40 years old (29.6% vs. 6.7%, p < 0.001) for progressive disease and for patients <40 years old for risk factors for sudden death (15.7% vs. 2.1%, p = 0.002). Patients with diffuse rather than confluent enhancement had two or more risk factors for sudden death (87% vs. 33%, p = 0.01). CONCLUSIONS: Gadolinium CMR reveals myocardial hyperenhancement in HCM. The extent of hyperenhancement is associated with progressive ventricular dilation and markers of sudden death. PMID- 12742299 TI - Gadolinium-enhanced magnetic resonance imaging in hypertrophic cardiomyopathy: in vivo imaging of the pathologic substrate for premature cardiac death? PMID- 12742300 TI - Effectiveness of implantable defibrillators for preventing arrhythmic events and death: a meta-analysis. AB - OBJECTIVES: The aim of this study was to compare the effectiveness of the implantable cardioverter defibrillator (ICD) and medical strategies for prevention of arrhythmic events and death. BACKGROUND: The ICD is a potential strategy to reduce mortality in patients at risk of sudden death. METHODS: The MEDLINE, EMBASE, and Cochrane Library electronic databases were searched from January 1966 to April 2002. All published randomized controlled trials comparing ICD implantation with medical therapy were reviewed. Four independent reviewers extracted data on all-cause mortality, nonarrhythmic death, and arrhythmic death using a standardized protocol. RESULTS: Nine studies including over 5,000 patients were synthesized using both fixed-effects and random-effects models. The primary and secondary prevention trials showed a significant benefit of the ICD with respect to arrhythmic death, with relative risks (RR) of 0.34 and 0.50, respectively (both p < 0.001). The mortality benefit of the ICD was entirely attributable to a reduction in arrhythmic death (all trials: p < 0.00001). Whereas the secondary prevention trials exhibited a robust decrease in all-cause ICD mortality (RR 0.75; p < 0.001), the pooled primary prevention trials demonstrated decreased all-cause ICD mortality (RR 0.66; p < 0.05) which was dependent on selected individual trials. The disparity in ICD-related mortality reductions in the primary prevention trials was related to variability in the incidence of arrhythmic death between individual studies. CONCLUSIONS: Although the ICD decreases the risk of arrhythmic death, its impact on all-cause mortality is related to the underlying risk of arrhythmia-related death relative to competing causes. Given the cost of the device strategy, policies of targeted intervention based on the future risk of arrhythmia are warranted. PMID- 12742301 TI - Value of rapid beta-blocker injection at peak dobutamine-atropine stress echocardiography for detection of coronary artery disease. AB - OBJECTIVES: We studied the value of a rapid beta-blocker injection at peak dobutamine-atropine stress echocardiography (DASE) for the detection of coronary artery disease (CAD). BACKGROUND: The presence of tachycardia and hyperdynamic wall motion may make it difficult to recognize a new wall motion abnormality (NWMA) at peak stress. METHODS: We studied 101 patients (mean age 58.2 +/- 9.8 years) who underwent effective DASE and coronary angiography. All patients received a rapid intravenous injection of metoprolol immediately after peak DASE image acquisition. Positivity in combined peak plus post-metoprolol images was defined when there was only peak NWMA, maintenance of peak NWMA, or NWMA detected only after metoprolol injection. Significant CAD was defined as >or=50% stenosis by quantitative angiography. RESULTS: There were 37 patients without and 64 with CAD. The sensitivity, specificity, accuracy, and positive and negative predictive values for the detection of CAD at peak stress were 84%, 92%, 87%, 95%, and 77%, respectively. Five patients with CAD had negative peak images that became positive only after metoprolol. Extension of peak NWMA during metoprolol was observed in 14 patients, and multivessel CAD was detected in 10 of them. The sensitivity, specificity, accuracy, and positive and negative predictive values for peak plus metoprolol images were 92%, 89%, 91%, 94%, and 87%, respectively. CONCLUSIONS: The use of metoprolol injected at peak of dobutamine infusion improved the detection of CAD by DASE. PMID- 12742302 TI - Diastolic mitral annular velocity during the development of heart failure. AB - OBJECTIVES: We sought to investigate the mechanism of reduced diastolic mitral annular velocity with diastolic dysfunction, despite elevated left atrial (LA) pressure. BACKGROUND: The peak rate of left ventricular (LV) early diastolic filling (E) and velocity of the mitral annulus due to long-axis lengthening (E(M)) are reduced in mild diastolic dysfunction. With more severe dysfunction, E increases in response to increased LA pressures. In contrast, E(M) decreases, despite increased LA pressure. METHODS: We studied eight dogs instrumented to measure LA pressure, LV pressure, and internal dimensions during the progressive development of heart failure (HF) produced by rapid pacing. RESULTS: Early diastolic filling decreased after four days of pacing from 114 +/- 32 to 88 +/- 22 ml/s (p < 0.05), but with more severe HF, it progressively increased to 155 +/ 32 ml/s (p < 0.05). In contrast, E(M) progressively decreased from 44 +/- 12 mm/s during the control period to 24 +/- 8 mm/s after four weeks (p < 0.05). Although E(M) was related to the time constant of LV relaxation (tau) (R(2) = 0.85), E was not. The latter occurred coincident with termination of the early diastolic LA to LV pressure gradient during all conditions. In contrast, with increasing HF, E(M) was progressively delayed after LA to LV pressure crossover by 37 +/- 12 ms (p < 0.05). The time from E to E(M) was related to tau (R(2) = 0.97). CONCLUSIONS: With slowed relaxation during the development of HF, E(M) is reduced and delayed so that it occurs after early, rapid filling. Thus, with slowed relaxation, E(M) does not respond to the early diastolic LA to LV pressure gradient, because it occurs when LV pressure is greater than or equal to LA pressure. PMID- 12742303 TI - Predictors of cardiac morbidity and related mortality in children with acquired immunodeficiency syndrome. AB - OBJECTIVES: The aim of this study was to determine the prevalence of cardiovascular dysfunction and its predictors in children with acquired immunodeficiency syndrome (AIDS). BACKGROUND: Cardiovascular manifestations are common among children with AIDS but may be clinically occult. METHODS: We reviewed the medical records, echocardiograms, electrocardiograms, and Holter monitor studies of 68 children with AIDS. We tested clinical and demographic characteristics at the time of AIDS diagnosis for their ability to predict serious cardiac events, death, and cardiac death. RESULTS: The median time from AIDS diagnosis to death or end of follow-up was 1.0 year (range, 1 week to 7.9 years). Nineteen patients (28%) experienced serious cardiac events after AIDS diagnosis. Of 43 patients who died, 15 (35%) had cardiac dysfunction. Multivariable analyses revealed that recurrent bacterial infections, wasting, encephalopathy, male gender, and an earlier year of AIDS diagnosis were predictors of serious cardiac events (relative risk [RR] = 9.3, 6.9, 4.7, 4.1, and 0.76, respectively, p < 0.05). Wasting, encephalopathy, a low age-adjusted CD4 count, a low age-adjusted immunoglobulin G (IgG) level, and an earlier year of AIDS diagnosis increased the risk of all-cause mortality (RR = 8.9, 5.1, 2.7, 0.82, and 0.8, respectively, p garenoxacin = grepafloxacin = levofloxacin = moxifloxacin = trovafloxacin (0.5 microg/ml) > ciprofloxacin = sparfloxacin (1 microg/ml). Gene sequencing of the quinolone resistance determining region and epidemiologic typing of 30 RSF isolates showed diverse mutational events in gyr A and par C and multiple pulsed-field gel electrophoresis (PFGE) patterns among strains that was not consistent with clonal dissemination. Continued surveillance by global or national networks should continue to monitor for H. influenzae isolates that are refractory to fluoroquinolone therapy. PMID- 12742321 TI - Geographic variations and trends in antimicrobial resistance among Enterococcus faecalis and Enterococcus faecium in the SENTRY Antimicrobial Surveillance Program (1997-2000). AB - In the 1990s, the Enterococcus emerged as an important pathogen because of increasing prevalence and acquired resistances to glycopeptides and other agents. The seriousness of this problem can vary markedly worldwide and within nations; the SENTRY Antimicrobial Surveillance Program documents these differences. Over 8,000 enterococci were processed in the program (1997-2000) and the Enterococcus faecalis (EF; 4,034 strains) and Enterococcus faecium (EFM; 1,123 strains) isolates are tabulated. All strains were processed by three regional monitors using reference dilution methods. Identification to species was performed by participants and confirmed by the central laboratories. EF occurrence was greater than EFM by ratios of 3:1 to 5:1 in the Asia-Pacific (APAC), European (EU), and North American (NA) regions; the ratio was 17:1 in Latin America (LA). EF and EFM represented approximately 80-90% of all isolated enterococci. Glycopeptide resistant enterococci (GRE) rates varied from nil for EF in APAC and LA to 43 to 54% in EFM in NA. A slight increase in GRE was noted in NA (EFM only). Van A phenotypes predominated all regions. The most recent (2000) rank order of % GRE by region was: for NA (13%) > LA (4%) > EU (3%) > APAC (1%). In NA potential therapeutic agents were (% S): ampicillin (81%), chloramphenicol (87%), quinupristin/dalfopristin (20%), ciprofloxacin (39%), gatifloxacin (51%), nitrofurantoin (83%) and linezolid (>99%). Resistances in enterococci continue to be documented worldwide, but rates within endemic areas like NA appears to be stabilizing. Van A resistance patterns predominate and therapeutic options continue to present dilemmas, although some of the older agents remain usable as primary therapy or as alternatives to the newer agents such as the oxazolidinones. PMID- 12742322 TI - Increasing the detection rate of group B streptococcal carriers by non-selective cervico-vaginal culture accompanied with the selective vagino-anorectal one. AB - It has been previously well defined that selective vagino-anorectal culture is required for detection of group B streptococcal (GBS) carriage. We interestingly observed that in 5.74% of carriers, GBS were recovered only from cervico-vaginal samples inoculated onto nonselective human blood agar, without recovery from vagino-anorectal samples inoculated to four selective media. PMID- 12742323 TI - Pseudooutbreak of Candida versatilitis fungemia in a microbiology laboratory. AB - Candida versatilitis was isolated from 10 blood cultures that had been supplemented with olive oil to promote the growth of Malassezia spp., and from the stock olive oil bottle in the laboratory. This unusual non-pathogenic yeast isolate was readily identified by DNA sequencing methodology. This report also points out that care must be taken to ensure the sterility of supplements added to blood culture media. PMID- 12742324 TI - Influence of patient age on the susceptibility patterns of Streptococcus pneumoniae isolates in North America (2000-2001): report from the SENTRY Antimicrobial Surveillance Program. AB - Risks of developing resistance in Streptococcus pneumoniae can be greatly influenced by antimicrobial use that is often dictated by patterns of prescribing at various patient ages. The results from the SENTRY Antimicrobial Surveillance Program (2000-2001; North America) were examined for differing susceptibility profiles (25 antimicrobials) indexed by five patient age groups. All MIC results were from reference methods performed in central laboratories. Pneumococcal resistance rates in the youngest patient group (0-5 years) were significantly (p < 0.05) higher for penicillin, other beta-lactams, macrolides, tetracyclines and trimethoprim/sulfamethoxazole. In contrast, fluoroquinolone resistance in S. pneumoniae was greatest in the patients >/= 65 years (1.4% resistance to gatifloxacin, levofloxacin and moxifloxacin), where use of this class has been escalating. Continued local and global surveillance of resistances in common respiratory tract pathogens such as S. pneumoniae appears to be prudent practice to guide effective chemotherapy. PMID- 12742326 TI - The discussion section. PMID- 12742327 TI - Evidence of iatrogenic HIV transmission in children in South Africa. PMID- 12742328 TI - Influence on fetal blood pH when adding amniotic fluid: an in vitro model. AB - OBJECTIVE: To assess the effect of contamination with amniotic fluid in different quantities on fetal capillary blood pH. DESIGN: In vitro model. SETTING: Department of Obstetrics, St Polten Hospital. SAMPLE: Venous umbilical blood and amniotic fluid from 35 women who underwent amniotomy during labour. METHODS: Venous umbilical blood was mixed in vitro with amniotic fluid in diluted series (venous umbilical blood/amniotic fluid, 10:1 to 1:1). In every case two parallel runs of the dilution series in an inverted fashion were performed to rule out a possible time-dependent bias of the pH measurements. MAIN OUTCOME MEASURES: pH change in dilutions of umbilical venous blood with amniotic fluid: 10:1, +0.07 (0.02); 8:1, +0.08 (0.02); 6:1, +0.08 (0.03); 4:1, +0.07 (0.02); 2:1, +0.09 (0.03); 1:1, +0.12 (0.05); all P < 0.001. RESULTS: Amniotic fluid pH increases, whereas umbilical blood pH decreases slightly over time since collection. Dilutions of umbilical venous blood with amniotic fluid resulted in a significant pH rise. There was no significant difference between the two inversely performed dilution series. CONCLUSION: Amniotic fluid influences in vitro fetal venous blood pH immediately after contact. This observation indicates the possible masking of a fetus in distress by fetal scalp blood pH determination when amniotic fluid contaminates the sample. PMID- 12742329 TI - Aggressive or expectant management of labour: a randomised clinical trial. AB - OBJECTIVE: To compare labour outcomes using aggressive or expectant management protocols. DESIGN: Randomised trial. SETTING: Pretoria Academic Complex, South Africa. It serves an indigent urban population. POPULATION: Healthy nulliparous women in active labour, at term, with a health singleton pregnancy and cephalic presentation. METHODS: The women were randomised to either aggressive (n = 344) or expectant (n = 350) management protocols. Aggressive management entailed using a single line partogram, a vaginal examination every two hours and use of an oxytocin infusion if the line was crossed. Expectant management entailed using a two line partogram, with the alert line and a parallel action line four hours to the right, with a vaginal examination every four hours. If the action line was reached, oxytocin was started. The women were reassessed every two hours thereafter. Analgesia was prescribed on request. MAIN OUTCOME MEASURES: Mode of birth, use of oxytocin and analgesia and neonatal outcome. RESULTS: The groups were similar with respect to maternal age, cervical dilation at trial entry, number crossing the alert line and birthweight of the infants. Significantly fewer women managed aggressively had caesarean sections (16.0%) than those managed expectantly (23.4%) (relative risk [RR] 0.68, 95% confidence intervals [CI] 0.50, 0.93). Significantly more oxytocin was used in the aggressive management group, but there was no difference with respect to the use of analgesia or episiotomy or in neonatal outcome with respect to the Apgar score at 1 or 10 minutes. There were three perinatal deaths. One woman was found to have an intrauterine death before trial entry and the other two were in the aggressive management group but did not receive oxytocin. Compliance by staff was poor in the aggressive management group. CONCLUSIONS: Aggressive management of labour reduces the caesarean section rate in nulliparous women but requires more intensive nursing. PMID- 12742330 TI - Factor V Leiden in pregnancies complicated by placental abruption. AB - OBJECTIVE: Recent studies suggest an increased prevalence of obstetric complications in female carriers of hereditary or acquired thrombophilias. The aim of the study was to determine if carriership of the factor V (FV) Leiden mutation (activated protein C [APC] resistance) is higher in women who have had of placental abruption during pregnancy. DESIGN: A retrospective case-control study. SETTING: University Hospital MAS, Malmo, Sweden. METHODS: A comparison of 102 women with placental abruption with 2371 prospectively collected controls. Carriership of FV Leiden was determined and the women were interviewed. MAIN OUTCOME MEASURES: Proportion of FV Leiden carriership, first degree heritage of thrombosis and previous placental abruption in cases and controls. RESULTS: Carriage of FV Leiden was found in 15.7% of women who have had placental abruption as compared with 10.8% of controls (P = 0.12, odds ratio [OR] = 1.5, 95% confidence interval [CI] = 0.9-2.7). Around 20% of women with placental abruption reported first degree heritage for venous thrombosis, as compared with 6.7% of controls (P < or = 0.001). CONCLUSIONS: FV Leiden carriership was not significantly different in women with placental abruption. However, there was an increased prevalence of first degree heritage for venous thrombosis in women with placental abruption, indicating a higher prevalence of thrombophilia among women with placental abruption. PMID- 12742331 TI - Dexamethasone and ursodeoxycholic acid protect against the arrhythmogenic effect of taurocholate in an in vitro study of rat cardiomyocytes. AB - OBJECTIVE: To establish whether the therapeutic agents ursodeoxycholic acid and dexamethasone protect cardiomyocytes from taurocholate-induced arrhythmias in an in vitro model. DESIGN: Laboratory study. SETTING: Imperial College London, Hammersmith Campus. SAMPLE: Neonatal rat cardiomyocytes. METHODS: Using scanning ion conductance microscopy, we measured the rate, rhythm, amplitude of contraction and calcium dynamics of ventricular myocytes from one to two day old rats. Cells were pre-incubated for 16 hours in dexamethasone (80 or 800 nM) or 0.1 mM ursodeoxycholic acid before adding taurocholate at different concentrations (0.3-4.5 mM). MAIN OUTCOME MEASURES: Changes in rate and amplitude of contraction, calcium dynamics and rhythm. RESULTS: Taurocholate at concentrations of up to 3 mM induces abnormal changes including reductions in rate, amplitude of contraction, abnormal calcium dynamics and dysrhythmias. Although dexamethasone had no immediate protective effect on these changes, pre incubation with dexamethasone was protective. Ursodeoxycholic acid pre-incubation was protective at taurocholate concentrations up to 1 mM. CONCLUSION: The therapeutic agents dexamethasone and ursodeoxycholic acid appear protective against the arrhythmogenic effect of taurocholate on cardiomyocytes. PMID- 12742332 TI - Aspirin (100 mg) used for prevention of pre-eclampsia in nulliparous women: the Essai Regional Aspirine Mere-Enfant study (Part 1). AB - OBJECTIVE: To reduce the incidence of pre-eclampsia in nulliparous women, in accordance with the suggestion of a recent meta-analysis that low dose aspirin might decrease this incidence by more than half if used early enough in and at a sufficient dose during pregnancy (more than 75 mg). DESIGN: Multicentre randomised double-blinded placebo-controlled trial. SETTING: Twenty eight centres in Northern of France and one in Belgium. POPULATION: Three thousand and two hundred ninety-four nulliparous women recruited between 14 and 20 weeks. METHODS: Randomisation to either 100 mg aspirin or placebo daily from inclusion through 34 weeks. MAIN OUTCOME MEASURES: Preeclampsia was defined as hypertension (> or =140 and or 90 mmHg) associated with proteinuria (> or =0.5 g/L). RESULTS: The aspirin (n = 1644) and placebo (n = 1650) groups did not differ significantly in the mothers' incidence of pre-eclampsia (28 of 1632 [1.7%] vs 26 of 1637 [1.6%]; relative risk, RR, 1.08, 95% CI 0.64-1.83), hypertension, HELLP syndrome or placental abruption, or in the children's incidence of perinatal deaths or birthweight below the 10th centile. The incidence of babies with birthweight below the third centile was significantly higher in the aspirin group, with no explanation. The incidence of maternal side effects was higher in the aspirin group, principally because of a significantly higher rate of haemorrhage. CONCLUSIONS: Aspirin at a dose of 100 mg does not reduce the incidence of pre eclampsia in nulliparous women. Aspirin (100 mg) is associated with an increase in bleeding complications. PMID- 12742333 TI - Randomised comparison of uterine artery Doppler and aspirin (100 mg) with placebo in nulliparous women: the Essai Regional Aspirine Mere-Enfant study (Part 2). AB - OBJECTIVE: To assess the effectiveness of a pre-eclampsia prevention strategy based on routine uterine artery Doppler flow velocity waveform examination during the second trimester of pregnancy, followed by a prescription for 100 mg aspirin in the case of abnormal Doppler findings. DESIGN: Multicentre randomised controlled trial. SETTING: Eleven centres in the north of France and one in Belgium. POPULATION: One thousand and eight hundred and fifty-three nulliparous women recruited between 14 and 20 weeks of gestation. METHODS: Randomisation either to undergo a uterine Doppler examination between 22 and 24 week of gestation or to take a placebo. Women with abnormal Doppler waveforms received 100 mg of aspirin daily from Doppler examination through 36 weeks. MAIN OUTCOME MEASURES: Pre-eclampsia was defined as hypertension (> or = 140 and/or 90 mmHg) associated with proteinuria (> or = 0.5 g/L). RESULTS: One thousand two hundred and fifty-three women (67%) were randomised into the systematic Doppler group and 617 (33%) into the placebo group. Of the 1175 patients in the Doppler group who underwent this examination, 239 (20.3%) had abnormal uterine artery Doppler and received a prescription for aspirin. Despite the aspirin prescription, the frequency of pre-eclampsia did not differ between the systematic Doppler group and the placebo group (28 of 1237 [2.3%] vs 9 of 616 [1.5%]; RR = 1.55, 95% CI 0.7-3.3). Furthermore, the groups did not differ in the frequency of children who were very small for their gestational age (< or =3rd centile) or for perinatal deaths. Compared with patients with normal Doppler findings, those with abnormal Doppler were at high risk of pre-eclampsia (RR = 5.5, 95% CI 2.5-12.2) and of giving birth to a small-for-gestational-age child (RR = 3.6, 95% CI 1.6-8.1). CONCLUSION: Despite its sensitivity in screening for pre-eclampsia, routine uterine Doppler in the second trimester cannot be recommended for nulliparous patients. PMID- 12742334 TI - Antenatal screening for human platelet antigen-1a: results of a prospective study at a large maternity hospital in Ireland. AB - OBJECTIVE: Fetomaternal mismatch for human platelet antigen (HPA)-1a accounts for approximately 85% of cases of neonatal alloimmune thrombocytopenia. The purpose of the study was to determine the prevalence of the HPA-1a negative platelet phenotype in a cohort of pregnant women in Ireland, the rate of alloimmunisation to HPA-1a in HPA-1 mismatched pregnancies and the associated incidence of neonatal alloimmune thrombocytopenia. DESIGN: A prospective case-control study. SETTING: The antenatal clinics of a large maternity teaching hospital. POPULATION OF SAMPLE: Pregnant women, regardless of parity, presenting at the antenatal clinics. METHODS: An enzyme-linked immunosorbent assay (ELISA) designed for simultaneous HPA-1a typing and antibody detection was used. Further analysis for HPA-1a alloantibodies was performed using commercial ELISA's (GTI PakPlus and Pak1) and the monoclonal antibody immobilisation of platelet antigens assay. Confirmation of serological typing was by the polymerase chain reaction technique using sequence-specific primers (PCR-SSP). MAIN OUTCOME MEASURES: The presence of the HPA-1a negative phenotype and its association with the development of maternal anti-HPA-1a and infant thrombocytopenia. RESULTS: Eighty-four of 4090 consecutive women enrolled in the study tested positive for HPA-1a in the screening ELISA. Confirmatory genotyping was performed on 67 women (representing 80% of the cohort), and 54 women (representing 3272 non-selected pregnancies), were homozygous for the HPA-1b allele (1.7%). Three of 34 (9%) women who delivered HPA-1a positive babies had detectable anti-HPA-1a and all three babies had neonatal alloimmune thrombocytopenia, for an overall incidence of 1:1100 non selected pregnancies. CONCLUSIONS: The observed prevalence of 1.7% for the HPA-1a negative platelet phenotype is as expected from studies in other countries. While we have demonstrated the practicability of antenatal HPA-1a screening, further research is warranted to investigate maternal parameters predictive of severe fetal thrombocytopenia in HPA-1a alloimmunised pregnancies. PMID- 12742335 TI - Case-control study of teratogenic potential of thiethylperazine, an anti-emetic drug. AB - OBJECTIVE: Thiethylperazine is a commonly used anti-emetic drug during pregnancy in Hungary. One experimental study in mice and rats found an increased occurrence of cleft palate after the use of thiethylperazine during pregnancy but the human data of thiethylperazine have not been reported. Thus, the aim of the study was to investigate the possible human teratogenic effect of thiethylperazine. DESIGN: Case-control approach. SETTING: The teratogenic potential of thiethylperazine was evaluated in the population-based large data set of the Hungarian Case-Control Surveillance of Congenital Abnormalities between 1980 and 1996. SAMPLE: Of 38,151 newborn infants without any congenital abnormalities (control group), 746 (2.0%) had mothers who were treated with thiethylperazine, while of 22,843 cases with congenital abnormalities, 411 (1.8%) had mothers who were treated with thiethylperazine during pregnancy. METHODS: Case-control pair analysis. MAIN OUTCOME MEASURES: Different congenital abnormalities. RESULTS: The pairs of cases with congenital abnormalities and their matched controls without congenital abnormalities were compared and this approach showed a somewhat higher rate of cleft lip +/- palate (OR: 2.0 with 95% CI: 1.0-4.0) in infants born to mothers with thiethylperazine treatment during the first trimester of pregnancy. CONCLUSION: Our data do not indicate clear teratogenic effect of thiethylperazine, however, a weak association was found between thiethylperazine use and cleft lip +/- palate. PMID- 12742336 TI - Pathways to evidence-based reproductive healthcare in developing countries. AB - OBJECTIVE: Developing country clinicians are aware of the discourse of evidence based medicine but heavily constrained in their ability to access and apply new knowledge. This study aimed to obtain primary descriptive data about access to and application of reproductive health-related medical knowledge in developing countries. DESIGN: Qualitative study using semi-structured interviews. SETTING: India and Yemen. SAMPLE: Forty-one doctors working in obstetrics, gynaecology, general practice and family planning services. METHODS: Interviews were tape recorded and transcribed verbatim. A theory-driven approach was conducted to carry out thematic analysis. MAIN OUTCOME MEASURES: Descriptions of experiences, attitudes and needs. RESULTS: Doctors felt the need to update their knowledge but this was not always achieved due to time and financial constraints. Alternative knowledge sources were described including the Internet, scientific meetings, medical family networks and speciality medical societies but access to these was limited by the expense of equipment, unfamiliarity with the technology and restricted advertisement of scientific meetings and conferences. Institutional hierarchy and conflict of generations were identified as barriers to change. Demoralisation was a common obstacle. On the positive side, involvement in medical education was a powerful driver for seeking new knowledge and applying research evidence to clinical practice. CONCLUSIONS: Priorities are to identify needs specific to different countries and medical settings in the developing world, access for all to full text journals and educational activities that fully engage practitioners in the early stages of their careers. Clinical teachers are the nodal group with the strongest incentive to obtain and use new knowledge. PMID- 12742337 TI - Radioimmunoscintigraphy with Tc-99m-labelled SM3 in differentiating malignant from benign adnexal masses. AB - OBJECTIVE: Ultrasound scanning, serum CA125 and menopausal status have previously been combined in a risk of malignancy index for the differential diagnosis of adnexal masses. Although this approach has greater accuracy than either individual tests or clinical assessment, it has a significant false positive and false negative rate. Efforts have been directed at refining differential diagnosis and this study assessed the role of radioimmunoscintigraphy using the stripped mucin 3 (SM3) antibody that has a 17-fold greater uptake in malignant than benign ovarian tumours in vitro. DESIGN: Prospective study of patients with a pelvic mass using radioimmunoscintigraphy. SETTING: Department of Nuclear Medicine of St Bartholomew's Hospital in collaboration with Cancer Network. POPULATION: A total of 93 patients with pelvic masses were recruited for this study of which 32 had ovarian cancer and 61 had benign lesions. METHODS: Radioimmunoscintigraphy was performed with Tc-99m-labelled SM3 (600 MBq), anterior and posterior pelvis imaged at 10 minutes and at 4 and 24 hours and evaluated with change detection analysis and probability mapping. MAIN OUTCOME MEASURES: Sensitivity and specificity of radioimmunoscintigraphy for ovarian cancer. RESULTS: Radioimmunoscintigraphy had a sensitivity for ovarian cancer of 84% (27 true positive and 5 false negatives) and a specificity of 87% (53 true negatives and 8 false positives) giving a negative predictive value of 91%. CONCLUSION: These results suggest that radioimmunoscintigraphy could be used to reduce the number of false positive findings in a strategy to refine differential diagnosis of the pelvic mass. PMID- 12742338 TI - Vaginal hysterectomy in nulliparous women without prolapse: a prospective comparative study. AB - OBJECTIVE: To compare vaginal hysterectomy success and complication rates in nulliparous and primiparous or multiparous women. DESIGN: A comparative prospective study. SETTING: Department of Gynaecology, La Conception Hospital, Marseille, France. POPULATION: Three hundred and forty-five consecutive patients without genital prolapse requiring hysterectomy for benign conditions and without previous pelvic surgery or caesareans were treated prospectively by vaginal hysterectomy. Fifty-two patients were nulliparous and 293 were primiparous or multiparous. METHODS: Data of patients were collected prospectively. MAIN OUTCOME MEASURES: Operative time, length of hospital stay, oophorectomy, conversion to laparotomy, intra- and post-operative complications. RESULTS: The mean operative time was significantly longer in nulliparous patients [95 (55.2) vs 79.9 (34.9) minutes, P < 0.01]. The overall complication rate was significantly higher [7/52 (13.46%) vs 13/293 (4.44%), P = 0.02, RR = 3.03 (1.27-7.23)] as was the haemorrhage rate in nulliparous compared with the primiparous and multiparous women [4/52 (7.69%) vs 5/293 (1.7%), P = 0.03, RR = 4.51 (1.25-16.23)]. Vaginal hysterectomy was successfully performed in 96.2% (50/52) of the nulliparous and 99.7% (292/293) of the parous patients [P = 0.06, RR = 1.04 (0.98-1.09)]. CONCLUSION: The success rate of vaginal hysterectomy in nulliparous women is high. However, the risk of complications seems higher than in the parous women. Laparoscopy assisted vaginal hysterectomy in nulliparous women needs further investigations. PMID- 12742339 TI - The pre-operative assessment of the adnexal mass: the accuracy of clinical estimates versus clinical prediction rules. AB - OBJECTIVE: To evaluate the reproducibility of the clinical judgement of gynaecologists, gynaecologists in training and gynaecologic oncologists and to compare the predictive performance of the offhand assessment with the predictive performance of existing mathematical models for the pre-operative assessment of the adnexal mass. DESIGN: Questionnaire with paper cases of women operated on for an adnexal mass. SETTING: Gynaecological unit in a teaching hospital in the South of The Netherlands. POPULATION: Women who underwent surgery for adnexal mass between January 1991 and December 1998. METHODS: We offered 45 gynaecologists five different sets of 34 cases, with data on female age and menopausal status, a written description of the sonography, Doppler flow measurement and serum CA125 measurement. Nine observers for every set were asked to estimate the probability of malignancy. MAIN OUTCOME MEASURES: The reproducibility of the risk estimates as made by the participants was expressed with an intraclass correlation coefficients. The accuracy of the judgement of the clinicians and the result of mathematical models in the prediction of malignancy were expressed with sensitivity, specificity, and receiver-operating characteristic curves. RESULTS: Neither clinically relevant nor statistically significant differences could be found between the accuracy of the risk assessments made by the clinicians and the accuracy of the risk assessments made by prediction models. CONCLUSION: This study demonstrates that at this moment there is no need to introduce complicated predictive scoring systems such as neural networks or logistic regression models for the pre-operative assessment of the adnexal masses. PMID- 12742340 TI - Antenatal erythropoietin and intra-operative cell salvage in a Jehovah's Witness with placenta praevia. PMID- 12742341 TI - Primary extranodal CD8 positive epitheliotropic T-cell lymphoma arising in a leiomyoma of the uterus. PMID- 12742342 TI - Neurofibromatosis type I and pregnancy: a fatal attraction? Development of malignant schwannoma during pregnancy in a patient with neurofibromatosis type I. PMID- 12742343 TI - A case of thrombotic thrombocytopenic purpura and neonatal alloimmune thrombocytopenia in the same pregnancy. PMID- 12742344 TI - Lumbosacral spondylodiscitis: an unreported complication of sacrocolpopexy using mesh. PMID- 12742346 TI - Recent developments in ocular gene therapy. AB - The past two to three years have witnessed a remarkable increase in the number of gene therapy studies to treat almost every disease of the eye. All types of delivery systems, viral and non-viral, have been used. Experiments have begun to move from the use of reporters, to genes with potential therapeutic value. In this paper, rather than giving an overview from the beginning of ocular gene therapy, I have chosen to review its most recent advances. Although numerous issues remain to be solved, the emerging picture is encouraging. Within the experimental setting, conditions in the anterior and posterior segments have been improved by the administration of genes encoding beneficial proteins. In one case, vision has been restored in a congenitally blind animal. Limitations do exit, however a greater understanding of the molecular biology of eye tissues coupled with the development of low immunogenicity vectors will continue edging the way for a future use of gene therapy in the clinical setting. PMID- 12742348 TI - Tight junction-related protein expression and distribution in human corneal epithelium. AB - PURPOSE: To investigate the expression and cellular distribution of the tight junction-related proteins occludin, claudin and ZO-1 in human corneal epithelium. METHODS: Light and electron immunohistochemistry was used to determine tissue distribution of occludin, claudin-1 and ZO-1 in the human corneal epithelium. Reverse transcription-polymerase chain reaction was used to reveal claudin mRNA expression in human corneal epithelium. RESULTS: In transverse sections, occludin and ZO-1 were localized at the apical cell-cell junctions between superficial cells in stratified corneal epithelium. Both basal and basolateral membranes of superficial cells were stained by the claudin-1 antibody, but no apical membrane staining was observed. In en face sections, claudin-1 and ZO-1 antibodies showed as bands that corresponded to the junctional complex. Claudin-1 staining of superficial cell cytoplasm was also observed. Occludin staining was seen at the junctional complex, where it was not continuous, but dotted along the cell junctions. The transcripts for claudin-1 and several other claudin isotypes, such as -2, -3, -4, -7, -9 and -14 were identified. CONCLUSION: Not only occludin, but also some claudins act as integral transmembrane proteins in the corneal epithelium. ZO-1 is a component of the corneal epithelial tight junction, as it is in most epithelial cells. PMID- 12742347 TI - Microphthalmia transcription factor analysis in posterior uveal melanomas. AB - PURPOSE: The protein encoded by the Microphthalmia gene (MITF) is a transcription factor essential for the development and survival of melanocytes. It serves as a master regulator in modulating extracellular signals. Because of its central role in melanocytes survival and to assess its potential use as a histopathological marker for melanoma, MITF expression was examined in human choroidal melanomas. METHODS: Fifty-seven paraffin-embedded sections of choroidal melanoma specimens and 1 choroidal melanoma cell line were analyzed using immunochemistry and RT PCR. Normal choroids and normal choroidal melanocyte cells were used as control. RESULTS: Sixty-five percent of the tumoral specimens stained positively for MITF with a predominant nuclear pattern of reactivity. MITF-M and MITF-A isoforms were detected by RT-PCR in all specimens examined. Using a chimeric protein resulting from the fusion of each Mitf protein with the GFP, Mitf-M exhibited an exclusive nuclear staining whereas Mitf-A exhibited a mixed nuclear and cytoplasmic staining. No correlation between MITF-positivity and parameters such as cell type, largest tumor diameter, sclera invasion, mitotic figures was observed. In contrast, a significant negative association was found between MITF staining and the pigmentation (p=0.02) and a positive correlation between MITF staining and the proliferative marker Ki67 was found (p=0.02). CONCLUSION: MITF may be implicated in choroidal melanoma pigmentation and proliferation. Further analysis should provide new insights into the mechanisms underlying the molecular and cellular changes of choroidal melanomas. PMID- 12742349 TI - Corneal thickness and endothelial cell density measured by non-contact specular microscopy and pachymetry in Rhesus macaques (Macaca mulatta) with laser-induced ocular hypertension. AB - PURPOSE: Sustained increase in intraocular pressure (IOP) in humans results in a loss of corneal endothelial cells and an increase of corneal thickness. The effects of chronically elevated IOP on the corneal endothelium of monkeys with laser-induced ocular hypertension, a commonly used animal model of human glaucoma have not been documented. This study examined the central corneal thickness (CCT), the corneal endothelial cell density (ECD), and the corneal endothelial cell size (ACS) in Rhesus monkeys with experimental ocular hypertension. Materials and methods. Ten male monkeys with argon laser-induced ocular hypertension in one eye for an average duration of 2.4+/-0.7 years, were sedated with ketamine hydrochloride, and the CCT, ECD, and ACS measured at the center of the cornea of both eyes with a Topcon SP-2000P non-contact specular microscope (Topcon America Corporation((R)), Paramus, NJ, USA). CCT was also measured using a DHG-500 Pachette ultrasonic pachymeter (DHG Technology Inc., Exton, PA, USA). Mean and standard deviation (S.D.) of CCT, ECD and ACS for each eye was calculated and statistically compared.Results. Mean CCT in the hypertensive and normal eyes measured by specular microscopy was 0.477+/-0.023mm and 0.468+/-0.020 mm, respectively. Mean ECD in the hypertensive and normal eyes was 2601.7+/-631.8 and 3990.2+/-402.9 cells mm(-2), respectively. The mean size of the endothelial cells was 252.4+/-23.9 micro m(2) in the normal eye and 408.7+/-115.0 microm m(2) in the hypertensive eye. No significant difference in the measurement of CCT was observed between the specular microscope and the pachymeter (p=0.46). No significant difference in the mean CCT was observed between the two eyes (p=0.4820), whereas the mean ECD was significantly lower in the hypertensive eye than in the normal eye (p<0.001). The ECD was inversely related to the length of IOP elevation (p<0.001). CONCLUSIONS: No difference in the corneal thickness measurement was observed between the specular microscopy and the pachymetry techniques. Chronic ocular hypertension did not significantly affect the CCT, but caused a significant loss of endothelial cells in the center of the cornea of the laser treated eyes compared to the normotensive eyes. The duration of elevated IOP was the most important factor affecting the ECD. PMID- 12742350 TI - The effect of roscovitine on herpetic keratitis. AB - To determine the effect of roscovitine, a potent antiviral in tissue culture administered intramuscularly to rabbits or by eye drops to mice for the treatment of herpetic keratitis this study was commenced.New Zealand white rabbits infected with McKrae strain herpesvirus (HSV-1) were treated twice a day with 10mg Roscovitine or vehicle from day 3 to 7, or 1% trifluridine eye drops five times a day. Severity of keratitis was graded daily by a masked observer. ICR strain mice were randomized into 14 groups. Both corneas of the mice were scarified with a 25 gauge needle, and were inoculated with the KOS strain of HSV-1. Roscovitine was dissolved in Cremophor((R)) and tissue culture medium. Group 1A, 1B, 2A, and 2B mice were treated eight times daily with either 800 micro M Roscovitine or its vehicle. Trifluridine treated animals (groups 3A and 3B) received topical treatment five times daily. In groups 1A-3B, the treatment was begun two days prior to or two days after viral inoculation. Mice were examined on days 2, 3, 5, and 9 after infection. In rabbits, the severity of keratitis in animals treated with intramuscular roscovitine was not significantly different from that in vehicle-treated animals except on day 7 (p=0.0460). In mice, there was no significant difference between roscovitine and vehicle treatment at any time point studied. However, the trifluridine treated mice had significantly lower scores compared to the roscovitine or vehicle-treated mice. Although roscovitine dramatically suppresses viral replication in tissue culture studies, in vivo this drug failed to alter the course of HSV keratitis in rabbits or mice. Considering high cost of roscovitine and the poor efficacy in these experiments, we feel that roscovitine is not feasible antiviral agent for today. PMID- 12742351 TI - Structural and cellular architecture of conjunctival lymphoid follicles in the baboon (Papio anubis). AB - Conjunctival lymphoid follicles (CLFs), present in normal individuals, undergo hyperplasia upon conjunctival infection by a specific array of pathogens; infection-associated enlargement of draining preauricular lymph nodes suggests that conjunctival follicles participate in the afferent limb of acquired immune responses for the ocular surface. The present study was performed to delineate the structural and lymphoid anatomy of CLFs in the baboon (Papio anubis), a non human primate conjunctival model with close similarity to the human. Conjunctiva from both eyes, along with mesenteric lymph node, spleen, tonsil, and ileum controls were harvested from ten baboons at necropsy, and studied by histochemical and immunohistochemical methods. Baboon conjunctival follicles were identified as dense oval collections of leukocytes in the substantia propria with infiltration into a thinned overlying conjunctival epithelium. Goblet cells were universally absent, the overlying mucin layer was attenuated, and the follicle associated epithelium (FAE) demonstrated comparatively diminished alkaline phosphatase expression. The basement membrane overlying each follicle appeared discontinuous. CD4-positive T lymphocytes were distributed in parafollicular areas and to a lesser degree in follicle germinal centers. B lymphocytes formed the predominant cell in follicles, and also heavily infiltrated the FAE. B cell IgM expression was prominent in germinal centers, while IgD staining occurred in a horseshoe-shaped distribution in the follicle mantle zone. Although B cell IgA expression was noted in the non-follicular conjunctiva, IgA expression was inconspicuous within conjunctival follicles. S-100- and CD1a-positive dendritic cells were found in FAE, while fascin-positive mature dendritic cells appeared in the deeper areas of each follicle. CD68-positive macrophages were dispersed throughout the follicles. CD35-positive follicular dendritic cells were observed only in germinal centers. CLFs appear highly organized consistent with a role in the adaptive immune response to conjunctival pathogens. PMID- 12742352 TI - Increased light damage susceptibility at night does not correlate with RPE65 levels and rhodopsin regeneration in rats. AB - The susceptibility of rats to light-induced retinal degeneration is increased at night. In mice, an important determinant of light damage susceptibility is the efficacy of rhodopsin regeneration after bleaching. The rate of rhodopsin regeneration is at least partly controlled by RPE65, a protein expressed in the retinal pigment epithelium. We therefore tested a potential involvement of RPE65 and rhodopsin regeneration in the increased light damage susceptibility of rats at night. For this purpose, rats were exposed to visible light at noon or at midnight and extent of light damage was determined by retinal morphology and TUNEL staining. Rpe65 gene expression was analyzed by semiquantitative RT-PCR and levels of RPE65 protein were determined by Western blotting. Rhodopsin regeneration kinetics was determined by measuring rhodopsin content immediately after a strong bleach and after different times of recovery in darkness. Rats were more susceptible to light damage at night as described by Organisciak and collegues [Invest. Ophthalmol. Vis. Sci. 41 (2000) 3694]. Rpe65 gene expression followed a day-night rhythm with highest steady-state mRNA levels at the beginning and lowest levels at the end of the day period. However, RPE65 protein levels remained constant. Rhodopsin regeneration kinetics did not differ during day and night. We conclude that levels of RPE65 protein and rhodopsin regeneration kinetics do not correlate with the increased light damage susceptibility observed in rats at night. Additional genetic or physiologic modifiers may exist in rats that regulate the retinal responsiveness to acute light exposure. PMID- 12742353 TI - Differential effects of aging on transport properties of anterior and posterior human sclera. AB - The transport properties and composition of 44 pairs of human sclera, 37-91 years were compared. Solute transport, diffusion and partition coefficients of posterior sclera for solutes ranging in mass from 0.023-70kDa were higher than those of anterior sclera; the posterior region was also more hydrated. The differences in partition coefficient between anterior and posterior sclera became more pronounced as solute molecular weight increased. Partition coefficients and hydration of both regions decreased with increasing age. Chondroitinase ABC digestion, which removed the majority of glycosaminoglycans, increased partition coefficients of both regions significantly. These results suggest that for regions of equal scleral thickness, neglecting the influence of vascular factors, drug delivery will be more readily achieved across the posterior sclera than the anterior sclera in the age group studied and that, for both regions, ease of delivery will decrease with decreasing age. PMID- 12742354 TI - Modulation of the Pasteur effect in retinal cells: implications for understanding compensatory metabolic mechanisms. AB - The purpose of the present experiments was to enhance understanding of the factors that are critical for the survival of retinal cells exposed to mitochondrial inhibition. Confluent cultures of Muller cells (rMC-1) and human retinal pigment epithelial cells (hRPE) were incubated in Dulbecco's minimal essential medium in the presence and absence of 1x10(-5)M Antimycin A, an inhibitor of mitochondrial electron transport. To modulate the rates of aerobic and anaerobic glycolysis, cells were incubated in media containing varying concentrations of glucose and 1-100 micro M of iodoacetic acid (IAA), an inhibitor of glyceraldehdye-3-phosphate dehydrogenase (G3PDH). Measurements were made of G3PDH, lactic acid production, and cellular ATP levels, along with an examination of cellular morphology, the latter providing an index of cellular viability. Control rMC-1 and hRPE produced lactate aerobically, respectively, at 0.48 and 1.50 micro molhr(-1)/10(6) cells. Anaerobically, lactate production increased 2-fold in rMC-1 and 3-fold in hRPE. Anaerobic ATP levels in both types of cells were maintained at control levels over 8hr. Experimental conditions were sought that would modulate only the capacity of rMC-1 and hRPE to increase glycolysis following mitochondrial inhibition, i.e. alter their Pasteur effect. We used low concentrations of IAA to partially inhibit G3PDH. Incubation of rMC-1 with IAA for 6hr caused a graded inhibition of G3PDH: 70% inhibition with 1 micro M, 90% with 5 micro M, 97% with 10 micro M, and 100% with 100 micro M. While the aerobic and anaerobic rates of lactic acid production were not altered by 1 micro M IAA, both were suppressed completely by 100 micro M IAA. However, incubation of rMC-1 with 5 micro M IAA caused a decrease of 30% in the rate of anaerobic lactic acid production but no change in the rate of aerobic glycolysis. Moreover, with 5 micro M IAA, rMC-1 incubated aerobically maintained ATP levels, but anaerobic ATP content decreased to a low level and cell morphology and viability were compromised. Essentially similar results were observed with hRPE. Both rMC-1 and hRPE are remarkably resistant to mitochondrial inhibition. This resistance is linked directly to the magnitude of the increase in the Pasteur effect. When the capacity of rMC-1 and hRPE to generate a Pasteur effect is selectively curtailed, these cells no longer are resistant to mitochondrial inhibition. It is suggested that in an intact tissue the ability of a cell to withstand a metabolic challenge will depend very much on the adequacy of the supply of glucose. Even a small limitation in the availability of this utilizable substrate and in the rate of the compensatory increase in the rate of anaerobic glycolysis could put the cell at greater risk during the challenge. PMID- 12742355 TI - Glycero- versus sphingo-phospholipids: correlations with human and non-human mammalian lens growth. AB - The human lens differs from other mammalian lenses in its very slow growth and unusual phospholipid composition of its cell membranes. Dihydrosphingomyelins (DHSMs) make up about half of all phospholipids in adult human fiber membranes. In all other membranes, sphingomyelins(SMs) with a trans double bond in their backbone, are prevalent. In our quest to understand the biological implications of such elevated DHSM levels, we analyzed membranes from various regions of human, elephant, giraffe, polar bear, pig and cow lenses. The levels of DHSMs were minor in non-human lens membranes. A strong correlation was observed between growth rate and relative contents of phosphatidylcholines(PCs) in epithelia and outer cortical fibers. Sphingomyelins became increasingly predominant in differentiated fibers and this increase was age dependent. Indeed, nuclear fiber membranes of aged non-human mammals were composed, almost exclusively, of (SMs). Although human lens membranes followed comparable compositional trends, the magnitude of the changes was much smaller. We postulate that the high relative contents of DHSMs provide a biochemically inert matrix in which only small amounts of PCs and SMs and their metabolites, known to promote and arrest growth, respectively, are present. This compositional difference is proposed to contribute to the slow multiplication and elongation of human lens cells. PMID- 12742356 TI - Comparison of ultrastructure, tight junction-related protein expression and barrier function of human corneal epithelial cells cultivated on amniotic membrane with and without air-lifting. AB - PURPOSE: To evaluate the usefulness of the air-lifting technique for culturing corneal limbal epithelial cells on amniotic membrane (AM) for use in ocular surface reconstruction. A cultured sheet that has a good barrier function should be better for this purpose. In corneal epithelium, tight junctions (TJ) play a vital role in the barrier function. The TJ complex includes the integral transmembrane proteins occludin and the claudins, and some membrane-associated proteins such as ZO-1. In this paper, we investigated the barrier function and the expression of TJ related proteins. METHODS: Corneal limbal epithelium obtained from donor corneas and cultivated on acellular AM was divided into two groups. These were the non-air-lifting (Non-AL) group, which was continuously submerged in medium, and the air-lifting (AL) group, which was submerged in medium for 3 weeks, then exposed to air by lowering the medium level. Morphology and the permeability to horseradish peroxidase (HRP) were determined by electron microscopy. Tight junction (TJ)-related protein and mRNA expression changes were assessed by immunoblotting and reverse transcription-polymerase chain reaction. RESULTS: The cultures of both groups formed 4-5-layer-thick, well-stratified epithelium. The AL cultures had tightly packed epithelial cells with all the HRP/diaminobenzidine (DAB) reaction product accumulated on the apical surface of the superficial cells. The Non-AL culture, by contrast, had more loosely packed epithelial cells with larger intercellular spaces. The HRP/DAB reaction product penetrated the intercellular space to a depth of 3-4 cell layers. Statistically, there was a significant difference in intercellular spaces and desmosome count in the superficial cells between the groups. With AL, TJ-related proteins localized at the apical portion of the lateral membrane. TJ-related protein and mRNA amounts were not changed by AL while claudin subtype expression became more consistent and closer to that of in vivo corneal epithelium. CONCLUSIONS: The AL technique reduces intercellular spaces in the superficial cells and promotes the formation of the barrier function. It is useful in culturing corneal epithelial cells for use in ocular surface reconstruction. PMID- 12742357 TI - Magnetic attraction of iron-endocytosed corneal endothelial cells to Descemet's membrane. AB - PURPOSE: To evaluate the feasibility of a novel method of magnetic attraction of iron-endocytosed corneal endothelial cells to Descemet's membrane. METHODS: Cultured rabbit corneal endothelial cells (RCEC) were exposed to spherical iron powder at various concentration ranging 0-100 micro moll(-1). After 24hr, the cell density and morphology were evaluated. RCEC that had been exposed to spherical iron powder (RCEC-iron), were trypsinized and poured onto a culture dish where a neodium magnet was fixated paracentrally. After 24hr, the cell density was measured at the areas with and without a magnet. Rabbits' corneas were cryo-injuried to detach corneal endothelial cells and 1x10(5)/200 micro l RCEC-iron were injected into the anterior chamber. Neogium magnet was fixed on the lid for 24hr to attract RCEC to Descemet's membrane. Each operated eye was observed up to 2 months after the injury. RCEC group (rabbits with cryo-injury and injection of normal cultured RCEC) and cryo group (rabbits with cryo-injury but without injection of RCEC) served as controls. RESULTS: The RCEC-iron density on the dish decreased in the medium containing iron powder of 10 micro moll(-1) or more. When RCEC had been exposed to iron powder of between 5 and 10 micro moll(-1), the ratio of RCEC in the field with a magnet to RCEC in the field without a magnet increased. In the RCEC-iron group, the mean corneal thickness gradually decreased and was significantly less than in the other two groups at 2, 4, and 8 weeks after the cell injection. Fluorescein microscopic examination showed a monolayer of DiI-labelled cells on the Descemet's membrane. CONCLUSION: Magnetic attachment of iron-endocytosed corneal endothelial cells to Descemet's nembrane can be a method of choice for corneal endothelial decompensation. PMID- 12742358 TI - Fate of cystatin C lacking the leader sequence in RPE cells. AB - The study made use of transiently transfected RPE cells expressing mature cystatin C (lacking the 26 amino acids leader sequence of the precursor cystatin C) fused to enhanced green fluorescent protein (EGFP) and, in control transfections, EGFP on its own. The findings demonstrate that 'leaderless' cystatin C is not processed through the secretory pathway of RPE cells. Since a polymorphism in the leader sequence has recently been associated with increased risk for development of exudative age-related macular degeneration (AMD), the present findings lend support to the hypothesis that impairment of function of the leader sequence may contribute to the aetiology of exudative AMD. PMID- 12742359 TI - Arsenic, cancer, and thoughtless policy. AB - The current drinking water contaminant standard for arsenic is 50 microg/L, as an upper bound. There is no lower bound for the standard. In an analysis of three epidemiology studies, the author demonstrates a significant cancer incidence trough for arsenic near 50 microg/L. Allowing, and even requiring, a much lower arsenic standard is not a desirable health outcome. Since most water systems currently deliver arsenic at levels below 5 microg/L, an undesirable health outcome is expected. For these systems and others not near 50 microg/L, the level of arsenic should be adjusted to bring it near 50 microg/L. PMID- 12742360 TI - Using rotifer population demographic parameters to assess impacts of the degradation products from trinitrotoluene phytoremediation. AB - The objective of this study was to evaluate the chronic lethal and sublethal aquatic toxicity effects associated with the phytoremediation of water contaminated with 2,4,6-trinitrotoluene (TNT) by the wetland plant species Myriophyllum aquaticum (parrot feather). Rotifers (Brachionus calyciflorus) feeding on an algal species (Nannochloropsis spp.) were used as the aquatic test organisms. Continuous flow laboratory microcosms were used to quantify effects on rotifer populations from TNT and the primary degradation product aminodinitrotoluene (ADNT) during and after phytoremediation. Rotifer demographic parameters from life tables, including survivorship, fecundity, reproductive values, net reproductive rate, generation time, intrinsic growth rate, and life expectancy, were used as measures of treatment effects. High-performance liquid chromatography analyses were performed to determine nitroaromatic concentrations. Results from this study have revealed significant differences in rotifer demographic parameters between microcosms with elevated initial TNT concentrations. Significant differences in demographic parameters also existed between the microcosms that did and did not receive phytoremediation treatment and the control microcosms. Study results have indicated that TNT phytoremediation via artificial wetlands not only may clean up hazardous waste at munitions sites but also may encourage the growth of aquatic populations such as rotifers. PMID- 12742361 TI - Effects of nicotine administration in developing muscle fibers of rats offspring. AB - The authors studied the effects of nicotine in developing skeletal muscles of rats. Pregnant Wistar rats received an enteral dose of nicotine (2.5, 5.0 or 10mg/kg, respectively, in groups G2, G5, and G10) from the 10th to the 18th or 20th days of pregnancy. Myotube atrophy was observed mainly in 20-day-old fetuses of G10. Twenty-day-old control fetus and of G2 and G5 had a high sarcoplasmic expression of desmin (weaker in G10). Vimentin expression was similar in 18-day old fetuses of the control, G2, and G5 groups, but it was weaker in 20-day-old fetuses of the G2 and G5 groups. This would indicate an acceleration of the maturation pattern of vimentin expression in these intoxicated fetus. In conclusion, high doses of nicotine induce myotube atrophy and decrease of the expression of intermediate filaments, whereas relatively low doses of nicotine (G2 and G5) induce an early decrease of vimentin expression with no myofiber atrophy. PMID- 12742362 TI - Biogenic arsenopyrite in holocene peat sediment, India. AB - The plant organic matter in the peat layer deposited around 6650+/-110 and 4800+/ 80 years BP, during the transgressive and regressive phases of sea level changes, respectively in the dried part (playa) of the present Pulicat lagoon in Palar Basin (southeast coast of India) was studied to elucidate the biogenic pyrite generation and associated trace elements. The scanning electron microscopic (SEM) observations show strongly curved unique C-shaped bacteria of uniform coccoidal shape and size (1 microm) freely scattered on the plant epidermal microfragments. These form spheroidal microcolonies 8-15 microm in diameter attached to the epidermis in a linear fashion or haphazardly enclosed in the translucent sheath as observed in surface view. These colonies are dense with a number of toxic trace elements as analyzed under an SEM-EDX spectrometer indicating its affinity toward metal binding. On maturity the sheath ruptures disseminating tetrad form of bacteria along with some C-shaped bacteria and few rhombohedral crystals of framboidal pyrite. The pyrite crystals intact in spheroidal shape in cavities on the epidermal fragments indicate its biogenic precipitation mediated by the bacteria. The pyrite crystals are rich in sulfur, iron, and traces of arsenic when analyzed by the SEM-EDX spectrometer. It is suggested that cubic and pyritohedran forms of crystals are perhaps arsenopyrite precipitated biogenically in saturated coastal wetland ecosystem in the past mediated by tetrad strain of the observed coccoid bacteria. Thus, the strata-bound peat layer could be the potent residing place for arsenopyrite or other toxic trace elements that may contaminate the groundwater aquifer. PMID- 12742364 TI - Carbon disulfide cytotoxicity on cultured cardiac myocyte cell of rats. AB - Although mostly epidemiological studies suggested that carbon disulfide produces cardiovascular effects in occupationally exposed workers, little is known about its cellular mechanism. The purpose of the present study was to evaluate the functional and histological effects on cardiac myocytes cultured under the condition of carbon disulfide exposure. Cardiac myocytes were isolated from neonatal rat ventricles by trypsin dispersion and cultured for 3 days in a full (Dulbecco's modified eagle medium) medium containing 2% calf serum. Thereafter the myocytes (10(6) myocytes/culture flask) were incubated with carbon disulfide at (CS(2)) the concentrations of 0, 20, 40, and 80 micromol/mL) for 24h. The beating arrest rate of myocytes for each group was examined and succinodehydrogenase (SDH) activity in the myocardial cells was also assessed by cytochemical method, and morphological examination was also performed. We found that the beating arrest rate of cardiac myocytes increased with increasing exposure levels. Vacuolization and pseudopodia may be seen in the cytoplasm of exposure group. SDH activity decreased with increasing exposure levels. The results suggested that CS(2) has a direct cytotoxic effect which is dose dependent. The biochemical mechanism may be a reduction of the availability of energy of the cardiac cytocyte in the form of ATP, resulting in a decrease of contractility by lacking of energy. PMID- 12742363 TI - Antioxidant responses and bioaccumulation in Ictalurus melas under mercury exposure. AB - Laboratory experiment was carried out to determine mercury accumulation in tissues (gills, kidneys, liver, and muscle) and biochemical responses in the liver of freshwater teleost Ictalurus melas. Catfish were subjected to different concentrations of Hg(2+) (35, 70, and 140 microg/L) for 10 days. The chemical analyses showed higher mercury concentrations for all treatments in gills and kidneys followed by liver and muscle. At the lowest mercury concentration a decrease in glutathione (GSH) content and an increase of GSH peroxidase Se dependent and glyoxalase II enzymes were observed. An increasing trend was observed also for GSH-S-transferase and glyoxalase I, while GSH peroxidase Se independent enzyme and GSH reductase showed no significant variation in activities. The increase in the enzymes activities of catfish, involved in the inactivation of reactive molecules formed during oxidative stress, could provide an additional protection against the oxidative damage induced by mercury. PMID- 12742365 TI - Effects of successive intrastriatal methylmercury administrations on dopaminergic system. AB - The present study was carried out in order to determine the effects of intrastriatal administration of different doses (40 microM, 400 microM, and 4mM) of methylmercury (MeHg) on dopaminergic system of rat striatum. Experiments were performed in conscious and freely moving rats using brain microdialysis coupled with liquid chromatography. Intrastriatal administration of MeHg produced significant increases in dopamine (DA) striatal levels (907+/-7%, 1870+/-319%, and 7971+/-534% for the doses of 40, 400 microM, and 4mM, with respect to basal). The increase in DA levels was associated with significant decreases in extracellular levels of its main metabolites dihydroxyphenylacetic acid (DOPAC) and homovallinic acid (HVA) (65.0+/-3.0% and 52.2+/-1.3%, respectively) using the dose of 4mM MeHg, whereas nonsignificant changes in metabolite levels were observed with the doses of 40 and 400 microM MeHg. A second infusion of 4mM MeHg 24h after first infusion also produced a rise of DA levels, but this increase was very small as compared with that produced by first infusion (7971+/-534% versus 985+/-186%). This second infusion of 4mM MeHg also decreased DOPAC and HVA levels, but this decrease was not significant as compared with that observed after first infusion (65.0+/-3.0% and 52.2+/-1.3% versus 62.4+/-5.2% and 63.4+/ 7.4%, respectively). We discuss these effects based on a stimulated DA release and/or a decreased DA intraneuronal degradation. PMID- 12742366 TI - The trace elements in response to lithium intoxication in renal failure. AB - The polyuric syndrome that develops as a consequence of chronic administration of lithium salts is most frequent and often causes complication in the treatment of manic depressive disease with the administration of drugs. It is known that kidneys play an essential role in systematic depositing of toxic metals. The purpose of this study was not only the determination of dose-dependent lithium concentration in serum and urine but also an estimation of sensitive biochemical indicators of nephrotoxicity detectable at an early stage after the administration of lithium carbonate to rats. Animals were given orally lithium salt to female Wistar rats at the dose of 10 and 20mg Li/kg daily during 5 weeks. In the urine diuresis protein concentration, copper, zinc, lithium and N-acetyl beta-glucoaminidase (NAG) activity were determined. In the serum also lithium, copper and zinc were analyzed. The results of the experiments indicate that the changes in urinary concentrations of essential copper, proteins, NAG activity and diuresis were observed when the concentration of lithium was ca. 9.79+/-1.68 mmol Li/L and in serum it corresponded to 0.3+/-0.06 mmol Li/L. These values corresponded to total doses of 150 mg Li/kg body weight administered to rats. In summary the increase of copper concentration, diuresis and urinary concentrations of protein and the NAG activity may be interpreted as a general metabolic response of kidneys induced by lithium detectable as an earlier indicator of nephrotoxicity. Therefore, regular determinations of lithium concentrations in serum of patients are important tools in the prevention of intoxication. PMID- 12742367 TI - Effect of three nitrophenols on carbon metabolism in Nostoc muscorum and Chlorella vulgaris. AB - Carbon metabolism in selected strains of Nostoc muscorum and Chlorella vulgaris grown in the presence of three nitrophenols [o-nitrophenol, m-nitrophenol and 2,4 dinitrophenol] was assessed by examining activities of the enzymes such as amylase, starch phosphorylase, fructose 1,6-biphosphatase, sucrose phosphate synthase, and invertase. Marked alterations were observed in activities of the enzymes involved in starch metabolism. The cellular content of starch in nitrophenol-grown cultures was significantly reduced, whereas the levels of nonreducing and reducing sugars significantly increased. There was a significant increase in the activities of amylase and phosphorylase, and these alterations are probably responsible for the decreased amount of starch in the cultures. Furthermore, significant changes were noticed in activities of the enzymes involved in synthesis of sucrose as well. PMID- 12742368 TI - The effect of mercury on the feeding behavior of fathead minnows (Pimephales promelas). AB - Fathead minnows (Pimephales promelas) were exposed to mercury (1.69, 6.79, and 13. 57 microg/L HgCl(2), 10 days exposure), and afterward their foraging ability was tested in a vegetated habitat for 7 days. Among the foraging metrics used were foraging efficiency, capture speed, and the ability to learn and retain information regarding habitat characteristics. In addition to behavioral tests, muscle tissue acetylcholinesterase activity and brain levels of several neurotransmitters were investigated. Comparisons with control fish and fish from the two highest exposure groups revealed consistent performance deficits in foraging efficiency and capture speed. However, no treatment effects on learning were detected, nor were differences in neurotransmitter levels detected. In determining the underlying proximate cause of the foraging deficits, it is believed that the greater pause time exhibited by treatment fish while foraging was the main cause of treatment differences. In the future, behavioral studies will continue to allow toxicity testing of environmentally relevant variables such as those used by behavioral ecologists. PMID- 12742369 TI - Effects of certain heavy metals on the growth, dye decolorization, and enzyme activity of Lentinula edodes. AB - Various physiological parameters of Lentinula edodes (Shiitake) in the presence of nine heavy metal salts were investigated. The mycelial growth was highly sensitive to cadmium and mercury, but less sensitive to zinc, copper, and lead. This resistance can be particularly dangerous to humans in the case of edible fungi such as Shiitake because of the possible heavy metal accumulation during growth and fruiting body production. All of the tested heavy metals inhibited decolorization of the dye Poly R-478 and the production of manganese peroxidase to a greater extent than they inhibited growth. Interestingly, with the exception of iron, the addition of all heavy metal salts investigated led to the increase of laccase production. Apart from cadmium and iron, none of the heavy metals inhibited the in vitro enzyme activities in concentrations up to 3mM. The results of this study indicated the applicability of L. edodes in biosorption technologies used in the removal of toxic metals from contaminated effluents and in bioremediation technologies designed to treat complex wastes contaminated with heavy metals in addition to other xenobiotics. PMID- 12742370 TI - Morphological abnormalities in Chironomus tentans exposed to cadmium-and copper spiked sediments. AB - The induction of mouthpart deformities and the developmental response with exposure to sediments spiked with three concentrations (9, 39, and 61 microgg(-1) Cd dry wt.) of cadmium (Cd) and three concentrations (30, 125, and 215 microgg( 1) Cu dry wt.) of copper (Cu) were investigated. Mouthpart deformity proportions in Chironomus tentans larvae were compared between metal-spiked and control populations and between parent and offspring (F1) populations. Cd- and Cu-treated sediments induced deformities (low Cd=13%, medium Cd=7%, high Cd=4%, low Cu=6%, medium Cu=9%, high Cu=6%) at significantly higher proportions than control (3%) sediments. No negative developmental response was determined. Larval sizes in metal-treated aquaria and control aquaria were not significantly different. F1 larvae from parents reared in medium and high Cu had significantly lower deformity rates than their parents. Our research adds to the growing evidence implicating heavy metals in general, and Cd and Cu specifically, as teratogenic agents. PMID- 12742371 TI - Monitoring of cadmium in the chain from soil via crops and feed to pig blood and kidney. AB - The relationships between cadmium (Cd) levels in soil, feed crops, feed concentrate, pig feed mixture, water, pig blood, and kidney from 49 farms were investigated and the possibility to use pig kidney as a bioindicator of available Cd in the agricultural environment was evaluated. There were correlations between Cd levels in soil and wheat, between wheat and barley, and between feed and kidney. The accumulation ratio between Cd levels in feed and kidney was on average 3. Animals from the same farm, raised in the same environment, given the same feed, and slaughtered at the same age had Cd levels in kidney and blood that could differ several times. This great variation, together with a considerable Cd contribution from nonlocally produced feed ingredients (concentrates), limits the possibilities to use Cd in pig kidney as an indicator of the available Cd in the local environment. PMID- 12742372 TI - Application of toxicity identification evaluation procedures to an effluent from a nitrogenous fertilizer plant in China. AB - The integrated method combining chemistry and toxicology, toxicity identification evaluation (TIE), was conducted to identify key toxicants in an effluent from a nitrogen fertilizer plant in China. Toxicity characterization, phase I of TIE, revealed that the suspected toxicant in the effluent was an anion that could be changed into a volatile acid. The results of toxicity identification and confirmation procedures indicated potassium cyanide to be the primary toxicant in the effluent. PMID- 12742373 TI - Effect of ammonia toxicity on the competition among three species of cladocerans (Crustacea: Cladocera). AB - Among the natural abiotic variables affecting the species density and composition of cladocerans, ammonia is important. Using population growth data as tool, we studied the competitive outcome of three cladoceran species (Ceriodaphnia dubia, Daphnia pulex, and Moina macrocopa) grown alone and together at three different concentrations (0, 25, and 100 mg L(-1)) of ammonia at one density (1 x 10(6)cells ml(-1)) of the microalga Chlorella vulgaris. Regardless of the presence or absence of competing species, C. dubia showed little population growth at 100 mg L(-1) of ammonia. At 25 mg L(-1) of ammonia, the population density of C. dubia increased but the value was much lower than that in control. D. pulex was able to grow at the ammonia concentration of 25 mg L(-1), comparable to controls. However, at 100 mg L(-1) of ammonia, the population declined and more rapidly when competed with other cladocerans. M. macrocopa showed decreased population density with increasing ammonia concentration in the medium. This trend was accelerated by the presence of competing cladocerans. M. macrocopa was benefited by the presence of D. pulex at 0 and 25 mg L(-1) of ammonia in the medium. The peak population density of C. dubia varied from 0.5 to 9 ind.ml(-1), whereas for D. pulex and M. macrocopa, these values varied from 1 to 3 ind.ml(-1) and 7 to 18 ind.ml(-1), respectively, depending on the ammonia concentration and the presence of competing species. The rates of population increase per day of C. dubia, D. pulex, and M. macrocopa ranged from -0.12+/-0.001 to 0.30+/-0.01, 0.04+/-0.001 to 0.22+/-0.002, and 0.201+/-0.028 to 0.235+/-0.019, respectively, depending on the ammonia concentration and the presence of competitors. Our study showed that while competition within the chosen cladoceran species caused suppression of one or two competing species, the presence of ammonia expedited this process based on the relative sensitivities of the tested zooplankton. PMID- 12742374 TI - Separation and toxicity of enantiomers of organophosphorus insecticide leptophos. AB - Enantiomers of leptophos were separated by high-performance liquid chromatography with a Whelk-O1 column using 3% dichloromethane in n-hexane as mobile phase. Toxicity tests of leptophos enantiomers and racemate were performed with daphnia. Enzyme inhibition of leptohpos was carried out by using butyryl cholinesterase from horse serum and acetylcholinesterase from housefly heads. From the inhibition test of butyrylcholinesterase, the half-inhibitory concentrations, IC(50), of (+)-leptophos, (-)-leptophos, and (+/-)-leptophos were 0.241, 1.17, and 1.05 gmL(-1), respectively. No significant difference in IC(50) in (-) leptophos and (+/-)- leptophos was found. However, the IC(50) of (+)-leptophos was significantly different from those of the others. In the inhibition test of acetylcholinesterase, the IC(50) values of (+)-leptophos, (-)-leptophos, and (+/ )-leptophos were 14.01, 24.32, and 13.22 gmL(-1), respectively. There was no significant difference in IC(50) in (+)-leptophos and (+/-)-leptophos, although the IC(50) of (-)-leptophos was significantly different from those of the others. From these results, leptophos-both enantiomers and racemate-seems to have higher neurotoxicity for mammals than for the target insects. In the toxicity test of daphnia, the half-lethal concentrations, LC(50), of (+)-leptophos, (-)-leptophos, and (+/-)-leptophos were 0.0387, 0.802, and 0.0409 gL(-1), respectively. There is no significant difference in LC(50) in (+)-leptophos and (+/-)-leptophos. The LC(50) of (-)-leptophos is significantly higher than those of the others. From these results, (-)-leptophos has lower toxicity to daphnia. PMID- 12742375 TI - First report and toxicological assessment of the cyanobacterium Cylindrospermopsis raciborskii from Portuguese freshwaters. AB - The freshwater cyanobacterium Cylindrospermopsis raciborskii has become increasingly prevalent in freshwaters worldwide. This species is a concern from a water quality perspective due to its known ability to produce a potent hepatotoxic alkaloid cylindrospermopsin, which has been implicated in outbreaks of human sickness and cattle mortality. C. raciborskii strains isolated from Brazil have also been found to produce the highly toxic paralytic shellfish poisons (PSPs). This article reports the toxicity of four strains of C. raciborskii taken from three reservoirs and one river in Portugal, as well as the occurrence of this species in other water bodies used for potable and recreational purposes. All four strains grown in pure culture in the laboratory were found to be toxic in the mouse bioassay at 8-24h after intraperitoneal administration of single doses ranging from 1337 to 1572 mgkg(-1) Histological examination indicated that liver damage was the primary lesion; in addition, there was inflammation in the intestine. HPLC/MS tests for the presence of cylindrospermopsin, microcystins, and PSP toxins were negative. The available evidence suggests that another toxin may be present. This constitutes the first report of toxic C. raciborskii in Europe and draws attention to the need for increased monitoring of this cyanobacterium in water bodies used for potable and recreational purposes. PMID- 12742376 TI - Immunopotentiation of intraepithelial lymphocytes in the intestine by oral administrations of beta-glucan. AB - Mice were orally administered with beta-glucan, isolated from baker's yeast, daily for one week (25mg/day/mouse) and several immunoparameters in the digestive tract were examined. The most prominent change was an increase in the number of intraepithelial lymphocytes (IEL) in the intestine, although the number of lymphocytes in the liver remained unchanged. The absolute number of both alphabetaT cells and gammadeltaT cells expressing CD8 antigens increased among IEL in the intestine. Primarily, liver lymphocytes showed a spontaneous production of Type 0 cytokine (simultaneous production of IFNgamma and IL-4) while IEL did not produce any cytokines without stimulation. However, mice administered with beta-glucan produced Type 1 cytokine, namely, production of IFNgamma alone. These results suggest that beta-glucan may be an important potentiator for mucosal immunity in the digestive tract. PMID- 12742377 TI - Treatment of adjuvant arthritis with granulocyte-colony stimulating factor and peptide derived from heat shock protein 65. AB - Adjuvant arthritis in Lewis rats is induced by the subcutaneous injection of Mycobacterium tuberculosis in mineral oil, and the predominant T cell immune reactivity is against the heat shock protein 65 derived peptide 176-190. We treated Lewis rats with human recombinant G-CSF followed by (i.v) administration of peptide 176-190 after induction of adjuvant arthritis (AA), and observed decreased disease severity, joint destruction, new bone formation and joint ankylosis. Treatment with G-CSF alone was also effective, but to a lesser extent. In addition, we found that splenocytes from rats treated with G-CSF had reduced antigen presenting capacity compared with splenocytes from vehicle treated rats. Primed lymph node cells from G-CSF plus peptide treated rats showed a marked reduction in proliferation and secretion of IFN-gamma after stimulation with the heat shock protein peptide in vitro as compared to controls. PMID- 12742378 TI - Impaired primary immune response in type-1 diabetes. Functional impairment at the level of APCs and T-cells. AB - We have recently described an impaired proliferative response of CD4(+) T-cells to primary antigens in patients with insulin-dependent diabetes mellitus (IDDM) [Clin. Immunol. 103 (2002) 249]. In order to further investigate possible mechanisms underlying this impairment, several factors known to be involved in the down-regulation of the immune response both at the level of APCs and CD4(+) T cells were investigated: Monocyte-derived dendritic cells (MDDC) from IDDM patients were shown to express elevated amounts of CD86 (B7.2) (p=0.003) and reduced amounts of the adhesion molecule CD54 (ICAM-1) (p=0.03) on their cell surface compared to age-matched healthy controls and patients with non-insulin dependent diabetes mellitus (NIDDM) as well as decreased SDS-PAGE stability of HLA-DQ and -DR peptide complexes directly isolated from the IDDM patients' peripheral blood mononuclear cells (PBMCs). Expression of CTLA-4 (CD152), known to be involved in the down-regulation of the immune response, was shown to be increased on CD4(+) T-cells from IDDM patients after exposure to the primary antigen KLH (keyhole limpet hemocyanin) presented by MDDC (p=0.0047). Likewise, purified CD4(+) T-cells from IDDM patients produced elevated levels of the cytokine TGF-beta1 after stimulation with immobilized monoclonal antibodies directed against CD3 and CD28 (p=0.014). When monocytes from IDDM patients were stimulated with lipopolysaccharide (LPS), an increased tendency to produce the inhibitory cytokine interleukin (IL)-10 (p=0.007) and the acute phase cytokine IL 6 (p=0.044) was observed, whereas the concentrations of tumor necrosis factor (TNF)-alpha, IL-1beta, and IL-12 were comparable to controls. Taken together, our data suggest that a deviation in the expression of certain molecules known to be involved in the peripheral control of the immune response is present in IDDM patients and is underlying the observed impairment of the primary immune response. PMID- 12742379 TI - U5A2-13, an antigen originally found on mouse NK-like T cells, is an early inducible cell surface antigen during lymphoid activation. AB - We have previously reported a monoclonal antibody (mAb), U5A2-13 mAb, which originally recognizes a phenotypically and functionally similar population of natural killer (NK)-like T cells. In this study, we found that U5A2-13 antigen (U5A2-13) was expressed not only on NK-like T cells but also on T and B cells during activation. In contrast to the low levels of U5A2-13 on freshly harvested T and B cells, the activation of these cells by various stimuli resulted in high levels of expression of U5A2-13 in vitro and in vivo. Similar to CD69, U5A2-13 is also expressed in most mouse lymphoid cell lines but not in nonhematopoietic cells. U5A2-13 on T cells reached maximal expression by 24h after stimulation and returned to baseline levels after 3 days. However, U5A2-13 differed from CD69 since its expression profile was different on CD4(+)- and CD8(+)-activated T cells, phorbol ester-activated EL-4 cells, and activated splenocytes in CD69 deficient mice. In addition, immunoprecipitation study indicated that U5A2-13 is not identical to CD69. Importantly, the U5A2-13-positive population of CD4(+) T cells exhibited significant levels of cytokine producing activity upon stimulation. Overall, U5A2-13 is an early inducible cell surface antigen that could be involved in lymphocyte activation. PMID- 12742380 TI - Activation of vascular endothelial cells by IL-1alpha released by epithelial cells infected with respiratory syncytial virus. AB - Although pulmonary inflammation is a serious, sometimes life-threatening, consequence of respiratory syncytial virus (RSV) infection, the mechanisms involved are not well understood. Since the process of inflammation is initiated by a complex series of events including the activation of specific adhesion molecules on vascular endothelium, we searched for endothelial cell-activating factors released from RSV-infected epithelial cells. We demonstrate here that vascular endothelial cells exposed to culture supernatants from RSV-infected pulmonary epithelial A549 cells are activated to express increased cell surface ICAM-1, and to a lesser extent, VCAM-1 and E-selectin. IL-1alpha was identified as the predominant endothelial cell-activating factor by pretreating epithelial cell supernatants with anti-IL-1alpha antibody. The preferential upregulation of endothelial ICAM-1 (relative to VCAM-1 and E-selectin) by RSV-infected epithelial cell supernatants was replicated by recombinant IL-1alpha thus confirming IL 1alpha as a major endothelial cell-activating cytokine released by RSV-infected epithelial cells. Il-1alpha mediated endothelial cell activation is thus a likely contributory event in the initiation of leukocyte inflammation associated with RSV infection. PMID- 12742381 TI - LPS up-regulates mucin and cytokine mRNA expression and stimulates mucin and cytokine secretion in goblet cells. AB - Bacterial inflammation in mucosa is accompanied by morphological and proliferative changes in goblet cells and mucin hypersecretion. Main stimulators of bacterial inflammation are bacterial lipopolysaccharides (LPS). In vitro investigation of the LPS effect on the molecular processes in goblet cells, using the human mucin-secreting goblet cell line HT29-MTX, showed the following results. LPS up-regulated mucin and cytokine mRNA expression and secretion in goblet cells in a concentration and time-dependent manner, with a maximum output at an LPS concentration of 100 ng/ml. LPS (100 ng/ml) increased mRNA expression of MUC5AC (2.4x), MUC5B (2.1x), and IL-8 (2.3x) and stimulated secretion of mucins (MUC5AC up to 39%, MUC5B up to 31%) and the inflammatory cytokine IL-8 (up to 10x). A significant correlation was found between the LPS-induced IL-8 secretion and secretion of mucins. These results suggest: (1) goblet cells, responding to the direct stimulation of bacterial LPS by two inflammatory-related processes such as production and secretion of the gel-forming mucins and the inflammatory cytokine IL-8, can be considered as an important part of mucosal immunity and (2) LPS- induced goblet cell mucin secretion can occur partly via IL 8-dependent pathway. PMID- 12742383 TI - Expression profiling of a transformed thymocyte cell line undergoing maturation in vitro identifies multiple genes involved in positive selection. AB - Biochemical and genetic studies of thymocyte maturation would be facilitated by the development of cultured cell lines that reflect stages of positive selection. We have derived a CD4(+)CD8(+)TCR(+) T-lymphoid cell line (M20) from a murine thymic tumor induced by a retrovirus carrying the v-myc oncogene (M-MuLV(myc)). M20 subclones undergo several aspects of positive selection in response to co culture with a thymic stromal cell line (St3), including down-regulation of CD4 and CD8, and up-regulation of CD5 and TCR. M20 possesses a functional TCR complex, and ligation of this complex produces changes similar to co-culture with St3 stroma. Expression profiling of M20 cells in this system identified 23 genes previously shown to be important in thymocyte maturation, as well as several novel candidate genes. This system provides a new model to elucidate the molecular mechanisms of thymocyte maturation and TCR-mediated cell signaling in double-positive thymocytes. PMID- 12742382 TI - Nitric oxide inhibits spleen cell proliferative response after burn injury by inducing cytostasis, apoptosis, and necrosis of activated T lymphocytes: role of the guanylate cyclase. AB - We previously showed that an overproduction of nitric oxide (NO) by macrophages was responsible for the collapse of lymphoproliferative responses after burn injury in rats. First, we demonstrate here that 10 days post-burn, the inhibition of splenocyte response to concanavalin-A results from cytostatic, apoptotic, and necrotic effects of NO on activated T cells. This was evidenced by various criteria at the levels of DNA, mitochondria, and plasma membrane. Inhibition of NO synthase by S-methylisothiourea (10 microM) normalized all the parameters. Second, we show that two soluble guanylate cyclase (sGC) inhibitors, LY83583 and ODQ, restored the proliferative response in a concentration-dependent manner. LY83583 (0.5 microM) rescued T cells from apoptosis. Similar results were obtained with KT5823 (5 microM) a specific inhibitor of protein kinase G (PKG). In contrast, neither LY83583 nor KT5823 inhibited NO-induced necrosis. These results suggest that NO blocked T cells in the G1 phase and induced apoptosis through a sGC-PKG-dependent pathway and necrosis through an independent one. PMID- 12742384 TI - Psychopharmacology research for individuals with mental retardation: methodological issues and suggestions. AB - Psychotropic medications are frequently prescribed for behavior problems and/or psychopathology among individuals with mental retardation (MR). Unfortunately, the methodological integrity of scientific studies that support the use of medications among this population is often lacking. A recent review of the literature revealed that many of the studies that have assessed the efficacy of psychotropic medications for individuals with MR are methodologically flawed. Thus, we have detailed suggestions to improve the quality of future medication studies and avoid the methodological problems that prevent the scientific advancement of psychopharmacological research among individuals with MR. PMID- 12742385 TI - Soles of the Feet: a mindfulness-based self-control intervention for aggression by an individual with mild mental retardation and mental illness. AB - Uncontrolled low frequency, high intensity aggressive behavior is often a barrier to community living for individuals with developmental disabilities. Aggressive behaviors are typically treated with psychotropic medication, behavioral interventions or their combination; but often the behaviors persist at a level that is problematic for the individual as well as care providers. We developed a mindfulness-based, self-control strategy for an adult with mental retardation and mental illness whose aggression had precluded successful community placement. He was taught a simple meditation technique that required him to shift his attention and awareness from the anger-producing situation to a neutral point on his body, the soles of his feet. After practice he applied this technique fairly consistently in situations that would normally have elicited an aggressive response from him. The data show that he increased self-control over his aggressive behaviors, met the community provider's requirement for 6 months of aggression-free behavior in the inpatient facility before being transitioned to the community, and then successfully lived in the community without readmission to a facility. No aggressive behavior was seen during the 1-year follow-up after his community placement. Mindfulness-based intervention may offer a viable alternative to traditional interventions currently being used to treat behavioral challenges in children and adults with mild mental retardation. PMID- 12742386 TI - Functional grouping in residential homes for people with intellectual disabilities. AB - The effects of functional grouping of people with intellectual disabilities on care practices in small residential homes in the community were investigated. A group comparison and a matched-pairs comparison were carried out in settings where less than or more than 75% residents were non-verbal, non-ambulant, had severe challenging behaviour, severe social impairment or were verbal and ambulant. Further analysis, focused on those with challenging behaviour was carried out using ordinal regression. In the group-comparison study, no significant differences were found for three of the five groups. Residents who were non-ambulant were rated as receiving care with less interpersonal warmth in grouped settings; residents with severe challenging behaviour were rated as receiving less good care practices in four respects (interpersonal warmth, assistance from staff, level of speech and staff teamwork) in grouped settings. The matched-pairs comparison found significant differences only for people with challenging behaviour, where grouped settings achieved less good results in terms of interpersonal warmth and staff teamwork. Higher adaptive behaviour and mixed settings were predictive of better care practices on 13 of 14 items of the Active Support Measure (ASM), with some setting variables also predictive for some items. Care practices only appear to vary for people with challenging behaviour, where grouped settings appear to offer less good results in some respects. PMID- 12742387 TI - Computer-based sorting-to-matching in identity matching for young children with developmental disabilities. AB - We evaluated a computer-based sorting-to-matching procedure to teach matching-to sample skills to seven young children with developmental disabilities who had failed to demonstrate identity matching-to-sample under the typical training procedure (such as observing a sample then selecting a comparison stimulus). In the sorting-to-matching procedure, rather than clicking on a comparison stimulus, the children moved the sample stimulus under the identical comparison stimulus. For all the children, identity matching-to-sample accuracy rapidly increased when the sorting-to-matching procedure was introduced, while it remained at chance levels in the typical training procedure. One of seven children showed collateral gains in accuracy with the typical training procedure after the exposure to the sorting-to-matching procedure. PMID- 12742388 TI - Increasing toy play among toddlers with multiple disabilities in an inclusive classroom: a more-to-less, child-directed intervention continuum. AB - We evaluated a more-to-less, child-directed continuum of interventions to increase toy play among toddlers with multiple disabilities in an inclusive classroom. In keeping with recommended practices in early intervention, all procedures were conducted within the toddlers' classroom. Following initial observations of three toddlers (under 3 years of age) that indicated toy play was less frequent than that of their typically developing peers, preference assessments were conducted of selected toys. The toddlers were then provided with repeated choices of preferred toys in a child-directed manner. Two of the toddlers subsequently received staff prompts and praise for toy play, representing a less child-directed (i.e., more staff-directed) intervention component, in addition to choices of preferred toys. Overall, toy play as well as nonprompted toy play increased for one toddler during the former condition and for two toddlers during the latter condition. For two of the toddlers, toy play increased to a level commensurate with that of their classmates who did not have disabilities. Small increases also occurred in the number of toys played with by each toddler. Results are discussed in terms of how intervention procedures can be applied along a more- to less-child-directed continuum based on individual child responsiveness to respective procedures. Areas for future research discussed include applying the continuum with other child behaviors, including more advanced toy play. PMID- 12742389 TI - Recidivism among offenders with developmental disabilities participating in a case management program. AB - This study examined recidivism, defined as arrests, among 252 clients who were accepted into a case management program for offenders with developmental disabilities. Overall, 40% of clients were arrested while participating in the program, and 34% were arrested within 6 months after case closure. The crimes for which clients were arrested tended to be minor: 21% were for probation or parole violations not associated with new criminal acts, 39% were for misdemeanors, 27% were nonviolent felonies, and 12% were for felonies against persons. Clients who completed the program (N=115) were less likely to be arrested after case closure than those who dropped out of the program (N=112), 25 and 43%, respectively. Other factors associated with arrests after case closure included having a developmental disability other than mental retardation, living in an urban area, being referred to the program by a criminal justice agency or through a private referral rather than a social service agency, and being arrested while in the program. Implications are discussed for service provision and evaluation of programs that work with offenders with developmental disabilities. PMID- 12742390 TI - Rhodopsin phosphorylation: 30 years later. AB - Phototransduction in vertebrate photoreceptor cells mediated by rhodopsin is one of the most comprehensively examined G protein-coupled receptor (GPCR) signaling pathways. The signal transduction pathway can be mapped from the initial absorption of light to conformational changes within rhodopsin, through activation of the G protein transducin, and to the ultimate closure of the cation cGMP-gated channels in the plasma membrane. Furthermore, phototransduction has become an intensely studied model system for understanding the desensitizing processes that allow reduced non-linear responses of photoreceptor cells to increasing levels of illumination. Although some general themes appear to occur in GPCR systems, the details of these desensitizing processes are likely to be specific to each of the receptors. These differences are attributed to the fact that each receptor has unique kinetic constraints, amplification levels, tolerance to basal constitutive activity, intracellular internalization and recycling, redundancy of isoforms, and morphologies of the cell of their expression. One of the biochemical processes that are believed to be a common part of this desensitization of the GPCR-mediated cascade is receptor phosphorylation catalyzed by members of a small family of the GPCR kinases. The enzymatic, physiological and genetic aspects of rhodopsin phosphorylation and rhodopsin kinase have been characterized extensively over the last 30 yr. However, new structurally based approaches to examining rhodopsin kinase and rhodopsin phosphorylation are still awaiting further investigations. We present here a summary of the current understanding of rhodopsin phosphorylation and the properties of rhodopsin kinase, along with some expectations of future investigations into these topics. PMID- 12742391 TI - Gene expression in the trabecular meshwork and the influence of intraocular pressure. AB - The trabecular meshwork (TM) tissue is responsible for maintaining the physiologic intraocular pressure (IOP) of the ocular globe. To perform this function the TM must rely on a variety of mechanisms. These mechanisms, acting either independently or in a coordinated manner, are governed by the expression of TM genes. Expression profiles of TM from adult intact tissue and infant cultured cells revealed the high level of diversity of the TM transcriptome, with only about 1% of its genes represented by more than 4 clones in any of the libraries. The profiles also revealed genes whose presence is associated with previously undescribed TM functions such the one that protects the TM tissue against calcification. These findings support the existence of numerous regulatory mechanisms in the TM and may help explaining the low percentage of glaucoma patients associated with each mutated glaucoma gene. Failure to maintain a physiological pressure can result in elevated IOP, a condition often associated with the development of glaucoma. Experimentally, different time-periods of an elevated pressure insult lead to the altered expression of distinct sets of genes. Thus, the ability of the TM to respond to mechanical and biochemical insults is possibly driven by induction or repression of a number of genes that, most likely, are different from those involved in regulation of normal IOP. None of the genes currently linked to glaucoma was present in the expression profile libraries whereas their expression in the TM was highly induced by effectors known to be causative of glaucomatous conditions. This analysis leads to the speculation that glaucoma candidate genes might be more related to genes responding to insults than to those involved in the maintenance of normal TM physiology. A recent study implicating the common stress mediator NF-kappaB in glaucoma would support this notion. Future library profiles utilizing distinct RNA sources together with differential expression studies between normal and glaucoma-triggering conditions and individual characterization of selected genes will help elucidate the relevant mechanisms for the regulation of IOP. PMID- 12742393 TI - Molecular determinants of retinal ganglion cell development, survival, and regeneration. AB - The retina is an easily accessible part of the CNS with a well-defined cytological architecture. It allows for detailed study of the regulation of neurogenesis, determinants of cell fate specification, and signals for cell survival versus programmed cellular death during development. Within the retina, retinal ganglion cells (RGCs) are the only neurons connecting to the brain. Their axonal projection to the midbrain targets, the superior colliculus (SC), and the lateral geniculate nucleus (LGN) has been subject of a number of investigations, and led to the identification of molecular signals directing topographic information for precise wiring during development. Transcription factors, guidance molecules, extracellular matrix proteins, neurotrophic factors, and cell death-regulating factors of the Bcl-2 family and caspases, have all been reported to be involved in the processes of formation of a precise retino-collicular map, and regulation of developmental cell death.During adulthood, RGCs and their projection have to be maintained, since-to our current knowledge-they cannot be replaced following injury. On the other hand, insults of various kinds can be potentially hazardous to RGCs. Therefore, much work has been directed towards understanding of the molecular regulation of RGC degeneration following insults such as retinal ischaemia, axonal lesion, or in optic neuropathy. Experimental strategies are being devised towards protection of lesioned RGCs. Since following axonal lesion, these cells not only need to survive, but also have to reconnect in order to be functionally relevant, efforts are directed towards not only survival, but also axonal regeneration and proper rewiring of injured RGCs. This paper reviews the molecular determinants of RGC fate determination and the development of the retino-tectal projection. We summarize what is known (and hypothesized) on the determinants of RGC survival during normal adulthood, and the mechanisms of RGC degeneration in the injured retina. We also try to develop perspectives towards neuroprotection and regeneration of adult lesioned RGCs that may be applicable to lesioned CNS neurons in vertebrates in a broader sense. PMID- 12742392 TI - Effects of retinal ganglion cell loss on magno-, parvo-, koniocellular pathways in the lateral geniculate nucleus and visual cortex in glaucoma. AB - Glaucoma is a leading cause of world blindness, and retinal ganglion cell death is its pathological hallmark. There is accumulating evidence that glaucomatous damage extends from retinal ganglion cells to vision centers in the brain. In an experimental primate model of unilateral glaucoma, degenerative changes are observed in magnocellular, parvocellular, and koniocellular pathways in the lateral geniculate nucleus, and these changes are presented in relation to intraocular pressure and the severity of optic nerve damage. Neuropathological findings are also present in lateral geniculate nucleus layers driven by the unaffected fellow eye. Finally, there is information on changes in the visual cortex in relation to varying degrees of retinal ganglion cell loss. The implications of these findings for refining concepts regarding the pathobiology of progression, and the detection and treatment of glaucoma, are discussed. PMID- 12742394 TI - Functions of insulin and insulin receptor signaling in retina: possible implications for diabetic retinopathy. AB - Insulin action regulates the metabolic functions of the classically insulin responsive tissues: liver, adipose, and skeletal muscle. Evidence also suggests that insulin acts on neural tissue and can modulate neural metabolism, synapse activity, and feeding behaviors. Insulin receptors are expressed on both the vasculature and neurons of the retina, but their functions are not completely defined. Insulin action stimulates neuronal development, differentiation, growth, and survival, rather than stimulating nutrient metabolism, e.g., glucose uptake as in skeletal muscle. Insulin receptors from retinal neurons and blood vessels share many similar properties with insulin receptors from other peripheral tissues, and retinal neurons express numerous proteins that are attributed to the insulin signaling cascade as in other tissues. However, undefined neuron-specific signals downstream of the insulin receptor are likely to also exist. This review compares retinal insulin action to that of peripheral tissues, and demonstrates that the retina is an insulin-sensitive tissue. The review also addresses the hypothesis that dysfunctional insulin receptor signaling in the retina contributes to cell dysfunction and death in retinal diseases. PMID- 12742396 TI - Risk assessment of chromium and arsenic in date palm leaves used as livestock feed. AB - Appreciable levels of total chromium (Cr) and arsenic (As) were found (by emission spectroscopy) in date palm leaves, which form a significant ingredient in livestock feed. The levels in the fruit were considered safe for human consumption. Our work involved evaluation of the distribution of these elements in the leaves during the developmental stages of the fruiting season. Thirty-six leaf specimens of the Fard cultivar were collected 9, 15 and 20 weeks, respectively, after pollination and subjected to a standard digestion procedure. Sample masses of typically 1 g (dry weight) were prepared in 25 ml dilute acid solution and investigated for trace levels of Cr and As by ICP-AES. Eleven soil samples collected at random during the growth stages were subjected to a similar digestion procedure and analysis. According to the literature, the permissible mean levels of Cr and As in plants are 200 and 80 ng/g (dry weight), respectively. We used these values as our guideline to assess the "risk" levels in our samples of interest. In the case of Cr, about 45% of the specimens possessed levels between 250 and 700 ng/g, while a significant number produced levels between 1000 and 5000 ng/g. With regard to As, about 70% of the samples were above the documented permissible mean value. As a result of the unusually high Cr concentrations in some cases, the feasibility of Cr "accumulation" in the leaves was examined. Insects and other organisms subsist on the leaves and an added concern was the accumulation of these elements in the food chain. The study formed an interesting contribution to environmental research, and the impact of our assessment on the environment is discussed. PMID- 12742397 TI - Investigating options for attenuating methane emission from Indian rice fields. AB - The development of methods and strategies to reduce the emission of methane from paddy fields is a central component of ongoing efforts to protect the Earth's atmosphere and to avert a possible climate change. It appears from this investigation that there can be more than one strategy to contain methane emission from paddy fields, which are thought to be a major source of methane emission in tropical Asia. Promising among the mitigating options may be water management, organic amendments, fertilizer application and selection of rice cultivars. It is always better to adopt multi-pronged strategies to contain CH4 efflux from rice wetlands. Use of fermented manures with low C/N ratio, application of sulfate-containing chemical fertilizers, selection of low CH4 emitting rice cultivars, and implementation of one or two short aeration periods before the heading stage can be effective options to minimize CH4 emission from paddy fields. Among these strategies, water management, which appears to be the best cost-effective and eco-friendly way for methane mitigation, is only possible when excess water is available for reflooding after short soil drying at the right timing and stage. However, in tropical Asia, rice fields are naturally flooded during the monsoonal rainy season and fully controlled drainage is often impossible. In such situation, water deficits during the vegetative and reproductive stage may drastically affect the rice yields. Thus, care must be taken to mitigate methane emission without affecting rice yields. PMID- 12742398 TI - Prediction of maximum daily ozone level using combined neural network and statistical characteristics. AB - Analysis and forecasting of air quality parameters are important topics of atmospheric and environmental research today due to the health impact caused by air pollution. As one of major pollutants, ozone, especially ground level ozone, is responsible for various adverse effects on both human being and foliage. Therefore, prediction of ambient ozone levels in certain environment, especially the ground ozone level in densely urban areas, is of great importance to urban air quality and city image. To date, though several ozone prediction models have been established, there is still a need for more accurate models to develop effective warning strategies. The development of such models is difficult because the meteorological variables and the photochemical reactions involved in ozone formation are very complex. The present work aims to develop an improved neural network model, which combines the adaptive radial basis function (ARBF) network with statistical characteristics of ozone in selected specific areas, and is used to predict the daily maximum ozone concentration level. The improved method is trained and testified by hourly time series data collected at three air pollutant monitoring stations in Hong Kong during 1999 and 2000. The simulation results demonstrate the effectiveness and the reliability of the proposed method. PMID- 12742399 TI - A comparative study of heavy metal concentration and distribution in deposited street dusts in a large and a small urban area: Birmingham and Coventry, West Midlands, UK. AB - Results are presented from a study of the distribution of heavy metals in street dusts of two cities in Midland England. The first (Birmingham) is a large urban area (population of 2.3 million), the second, Coventry, a small one (population of 0.3 million). Several trends were identified from Birmingham: higher concentrations were located near industrial areas in the northwest of the city and within the ring road. However, lower concentrations were found to the southwest in areas of mainly residential properties and parks. High values were also identified in association with junctions controlled by traffic lights where vehicles were likely to stop regularly. This last trend was further investigated in Coventry, where it was found that concentrations of heavy metals at junctions controlled by traffic signals and by pedestrian-controlled pelican lights (Mounted Pelican Controller, MPCs) were lower than those found in Birmingham, apart from Ni. PMID- 12742400 TI - Sorption of volatile organic compounds on typical carpet fibers. AB - Measurements of adsorption isotherms for three volatile organic compounds (VOCs) (toluene, 1,2-dichlorobenzene and 1,1,1-trichloroethane) on polyacrylonitrile carpet fibers over the temperature range 25-45 degrees C were carried out in a thermogravimetric analyzer (TGA). Linear isotherms were observed in all cases with values of the Henry coefficient ranging from 0.063 to 0.941 mm. The results of additional experiments carried out in a simple test chamber containing a single source of VOC showed that the carpet fibers acted as a significant sink causing a prolonged elevation of VOC concentration in the air within the chamber. An unsteady-state model is presented, which adequately described the adsorption and desorption phenomena occurring in the test chamber and yielded realistic values of the adsorption and desorption rate constants. There was good agreement between the equilibrium and kinetic constants obtained in the TGA and test chamber experiments. PMID- 12742401 TI - Organochlorine pesticide residues in sediments of a tropical mangrove estuary, India: implications for monitoring. AB - The paper examines the concentrations of isomers of hexachlorocyclohexane (HCHs), dichlorodiphenyl trichloroethane and its metabolites (DDTs), alpha-endosulfan and endosulfan sulfate in surface sediment samples collected from the mouth of Hugli estuary in the vicinity of Sundarban mangrove environment, eastern part of India. An overall pattern of accumulation of these pesticides was in the order of: SigmaHCH>endosulfan sulfate>SigmaDDT>alpha-endosulfan. The concentration of these compounds was quite low. An elevated level of SigmaHCH, SigmaDDT and endosulfan sulfate were marked during premonsoon months, a period characterized by high salinity and pH values. Among the isomers and metabolites of HCH and DDT, beta HCH, pp'-DDT and pp'-DDE were found to be dominant. The sources of contamination are closely related to human activities, such as domestic and industrial discharges, agricultural chemical applications and soil erosion due to deforestation. The study is compared to other estuarine environment in India and abroad. The present data will serve as a baseline against which future anthropogenic effects may be assessed. PMID- 12742402 TI - Evaluation of landfill leachate in arid climate-a case study. AB - Generation of leachate from municipal solid waste (MSW) landfill in arid regions has long been neglected on the assumption that minimal leachate could be formed in the absence of precipitation. Therefore, a case study was conducted at two unlined MSW landfills, of different ages, in the state of Kuwait in order to determine the chemical characteristics of leachate and examine the mechanism of leachate formation. Leachate quality data were collected from both active and old (closed) landfills where co-disposal of MSW and other solid and liquid wastes is practiced. The analysis of data confirms that leachates from both landfills are severely contaminated with organics, salts and heavy metals. However, the organic strength of the leachate collected from the old landfill was reduced due to waste decomposition and continuous gas flaring. A significant degree of variability was encountered and factors which may influence leachate quality were identified and discussed. A water balance at the landfill site was assessed and a conceptual model was presented which accounts for leachate generation due to rising water table, capillary water and moisture content of the waste. PMID- 12742403 TI - Risk assessment and pathway study of arsenic in industrially contaminated sites of Hyderabad: a case study. AB - Different areas in the industrial region of Patancheru near Hyderabad, Andhra Pradesh (A.P), India are contaminated with high concentration of arsenic, which is attributed to industrial source like veterinary chemicals, pharmaceuticals, pesticide industries, etc. Fourteen villages of this area of Patancheru were assessed for arsenic contamination by collecting samples of water (surface and ground), soil, fodder, milk, and vegetables. The total arsenic content in the whole blood, urine, hair, and nails of the residents showing arsenical skin lesions and other clinical manifestations were also studied. To understand the bioavailability of arsenic in this environment and its possible entry into human food chain, speciation studies of arsenic was carried out and the results are presented in this paper. PMID- 12742404 TI - Effects of organic acids on copper and cadmium desorption from contaminated soils. AB - A study was conducted to investigate the effect of organic acids on Cd and Cu desorption from natural contaminated soils (NCS) with permanent contamination by metal smelters and from artificial contaminated soils (ACS) derived from an artificial amendment of Cd to three representative zonal soils in Central China. Results showed that the desorption of Cd in either NCS or ACS, with the increment of tartrate or citrate concentration in desorption solution, can be characterized as a valley-like curve. The presence of tartrate or citrate at a low concentration (< or =0.5 mmol/l) inhibited Cd desorption from these two types of soils, whereas the presence of organic acids at high concentrations (> or =2 mmol/l for citrate and about > or =15 mmol/l for tartrate) apparently promoted Cd desorption. The desorption curve of Cu by tartrate solution with different tartrate concentrations can also be characterized as a valley-like curve, while the desorption of Cu in the presence of citrate was directly enhanced with the increment of citrate concentration. With the enhancement of initial pH value from 2 to 8 in the presence of citrate, Cu desorption ratio decreased at the first stage, then increased, and then decreased again. A valley and a peak sequentially appeared in the Cd or Cu desorption curve with initial pH value increment. Compared with citrate, the desorption ratio of Cd or Cu from NCS or ACS was directly decreased in the presence of tartrate, with the enhancement of the pH value from 2 to 8. Cd or Cu desorption was clearly enhanced when the electrolyte concentration of KNO3 or KCl increased in the presence of 2 mmol/l tartrate. Moreover, a higher desorption ratio of Cd or Cu was shown with KCl electrolyte than with KNO3 electrolyte with the same concentration. Based on these observations, we suggest that bioavailabilities of heavy metal can be promoted with selected suitable types and concentrations of organic acid amendment and reasonable field condition. PMID- 12742405 TI - Kinetics of trace element uptake and release by particles in estuarine waters: effects of pH, salinity, and particle loading. AB - The uptake and release of 109Cd, 51Cr, 60Co, 59Fe, 54Mn, and 65Zn were studied using end-member waters and particles from Port Jackson estuary, Australia. The kinetics of adsorption and desorption were studied as a function of suspended particulate matter (SPM) loading and salinity. Batch experiments showed that the position and slope of the pH edges are dependent on the metal and on the salinity of the water (except for Mn). The general effect of salinity was to move the adsorption edge to higher pH values, with the greatest change being found for Cd. Most of the metals showed relatively simple kinetics with an increase in uptake as a function of time and suspended particle concentrations. The time dependence of Cd uptake was more complex, with an initial adsorption phase being followed by strong mobilization from the suspended sediments, explained by chlorocomplexation and competition with seawater major cations. The reversibility of the sorption decreased in the order Co>Mn>Zn>Cd>Fe>Cr. The percentage of adsorbed metal released in desorption experiments was greater in seawater than freshwater for Cd, Zn, and Co. These results are important in understanding the cycling of pollutants in response to pH, salinity, and particle concentrations in estuarine environments. In addition, they give valuable insight into the important mechanisms controlling the partitioning of heavy metals in the Port Jackson estuary. PMID- 12742406 TI - Investigation of soil multi-element composition in Antalya, Turkey. AB - The chemical composition of 73 surface soil samples collected at the city of Antalya were analyzed for major, minor and trace elements to assess distribution of soil pollution, due to deposition of pollution-derived particles from the atmosphere. Comparison with data from rural area and distribution maps demonstrated that the composition of soil by metals is not significantly modified by anthropogenic activities in most of the city and its surroundings. In these areas, observed concentrations can be largely accounted for by occurrence of elements in aluminasilicate matrix of soil. However, soil composition is altered substantially close to major industries and at the settlement districts. In such limited areas, concentrations of anthropogenic elements such as Cd, Pb, Zn, Cu and Cr are factors of 20-50 higher than their concentrations in unperturbed soil. Factor analysis revealed three groups of elements that differ in their distributions. One of these components is unperturbed soil component, which is distributed uniformly in the study area; the second one is polluted soil, which is mostly confined to settlement areas and around industries; and the third is a mixed marine and motor vehicle impacted soil component, which occurred at the coastal parts of the city. PMID- 12742407 TI - Analysis of microcystins in cyanobacteria blooms and surface water samples from Meiliang Bay, Taihu Lake, China. AB - Taihu Lake is the third largest freshwater lake in China. In recent years, the water pollution of cyanobacteria blooms has become a severe problem in this area. Microcystins (MCs) are an important group of toxic compounds mainly produced by some cyanobacteria species and have both acute and chronic hepatotoxic effects on animals and humans. This paper presents the first data on the identification and detection of MCs in both natural occurring cyanobacteria blooms and surface water samples (0-0.5 m), collected from Meiliang Bay, Taihu Lake, China. A conventional method for extraction and isolation of MCs from cyanobacteria blooms was applied. High-performance liquid chromatography (HPLC) analysis showed that the main toxic component in the cyanobacteria materials was MC-LR. The monoclonal antibody (mAb) against MC-LR produced by hybridoma technique was employed for direct competitive ELISA to detect the concentrations of MCs in bloom and water samples collected in 2001. The results not only revealed the presence of MCs but also temporal variations of MCs levels of three sampling stations in Meiliang Bay in 1 year. It is obvious that the MC contents were relatively higher during warm months and related with the status of eutrophication. Our study indicates the threat associated with MCs in water body of Taihu Lake. To prevent the MCs potential hazard on public health in this area, some necessary measures of monitoring and control of growth of cyanobacteria are urgently needed. PMID- 12742408 TI - Urbanization, land use, and water quality in Shanghai. 1947-1996. AB - The paper undertakes a preliminary investigation into the relationship between water quality and urbanization as well as the changing patterns of land use within Shanghai. Longitudinal changes to water quality at various points along the course of the Huangpu River are analysed and compared to changes in the rates of urbanization and changes in land uses. The results reveal that rapid urbanization corresponds with rapid degradation of water quality. It also shows that urban land uses are positively correlated with the decline in water quality. A regression model shows that close to 94% of the variability in water quality classifications is explained by industrial land area. The paper concludes with the need for comprehensive land use planning as a way of protecting valuable water resources. PMID- 12742410 TI - TOR inhibitors and cardiac allograft vasculopathy: is inhibition of intimal thickening an adequate surrogate of benefit? PMID- 12742411 TI - Post-transplantation lymphoproliferative disease in heart and heart-lung transplant recipients: 30-year experience at Stanford University. AB - BACKGROUND: Post-transplantation lymphoproliferative disease (PTLD) is an important source of morbidity and mortality in transplant recipients, with a reported incidence of 0.8% to 20%. Risk factors are thought to include immunosuppressive agents and viral infection. This study attempts to evaluate the impact of different immunosuppressive regimens, ganciclovir prophylaxis and other potential risk factors in the development of PTLD. METHODS: We reviewed the records of 1026 (874 heart, 152 heart-lung) patients who underwent transplantation at Stanford between 1968 and 1997. Of these, 57 heart and 8 heart lung recipients developed PTLD. During this interval, 4 different immunosuppressive regimens were utilized sequentially. In January 1987, ganciclovir prophylaxis for cytomegalovirus serologic-positive patients was introduced. Other potential risk factors evaluated included age, gender, prior cardiac diagnoses, HLA match, rejection frequency and calcium-channel blockade. RESULTS: No correlation of development of PTLD was found with different immunosuppression regimens consisting of azathioprine, prednisone, cyclosporine, OKT3 induction, tacrolimus and mycophenolate mofetil. A trend suggesting an influence of ganciclovir on the prevention of PTLD was not statistically significant (p = 0.12). Recipient age and rejection frequency, as well as high dose cyclosporine immunosuppression, were significantly (p < 0.02) associated with PTLD development. The prevalence of PTLD at 13.3 years was 15%. CONCLUSIONS: The overall incidence of PTLD was 6.3%. It was not altered by sequential modifications in treatment regimens. Younger recipient age and higher rejection frequency were associated with increased PTLD occurrence. The 15% prevalence of PTLD in 58 long-term survivors was unexpectedly high. PMID- 12742412 TI - Donor PAI-1 expression inhibits the intimal response of early allograft vascular disease. AB - BACKGROUND: The development of allograft vascular disease (AVD) may be related to altered expression of the fibrinolytic system. We determined the extent to which plasminogen activator inhibitor type 1 (PAI-1) expression in donor tissue influences intimal proliferation (IP) in a mouse model of AVD. METHODS: We utilized an end-to-end abdominal aortic transplant model in mice to investigate the development of IP in 3 groups of 6 recipients. Group A (negative control) utilized C57BL/6J strain mice as both donors and recipients. In Groups B (positive control) and C, C57BL/6J mice were vessel donors and CBA/J mice were recipients. Both groups received intraperitoneal anti-CD4 and anti-CD8 monoclonal antibodies (250 microg/week for 5 weeks). Group C recipients, however, were transplanted with vessels from C57BL/6J PAI-1 knockout mice. Animals were killed at 50 days. Transplanted aortas were removed and intimal areas calculated using morphometric analysis. RESULTS: Group A (mean intimal area 6421 +/- 8507 microm(2)) demonstrated very little IP in comparison to the other groups. IP was significantly higher in Group B (mean intimal area 56357 +/- 35629 microm(2)) than Group A (p = 0.008). Group C (mean intimal area 288195 +/- 123279 microm(2)) demonstrated significantly more intimal proliferation than either Groups A or B (vs B, p = 0.003; vs A, p < 0.001). The significance of these results is maintained if intimal thickness is measured as a stand-alone reference for the intimal response. CONCLUSIONS: Lack of PAI-1 expression in donor tissue greatly exaggerates the extent of IP after allogeneic transplantation and suggests that PAI-1 is important in limiting the early phase of AVD. PMID- 12742413 TI - Early constriction or expansion of the external elastic membrane area determines the late remodeling response and cumulative lumen loss in transplant vasculopathy: an intravascular ultrasound study with 4-year follow-up. AB - BACKGROUND: Early constriction of the external elastic membrane (EEM) area has been observed after cardiac transplantation. The aim of this study was to compare the late disease process of transplant vasculopathy between coronary segments with early constrictive and expansive remodeling. METHODS: Serial intravascular ultrasound data obtained annually for 4 years after transplantation in 38 transplant recipients was available. In 135 matched segments from 59 coronary arteries ultrasound images were digitized at 1-mm intervals. Mean values of the external elastic membrane (EEM), lumen and intimal areas were calculated. On the basis of a decrease or increase in EEM area within the first year after transplantation, we defined segments with early constrictive remodeling (CR, n = 71) or early expansive remodeling (ER, n = 64). RESULTS: Annual changes in intimal area were similar between segments with early CR and ER throughout the follow-up period. However, during the second and third year, annual increases in EEM area were greater in segments with early CR than in segments with early ER (second year: 1.5 +/- 2.7 vs 0.6 +/- 2.8 mm(2), p = 0.052; third year: 1.3 +/- 2.5 vs -0.03 +/- 2.6 mm(2), p = 0.003). Despite this late expansion, segments with early CR showed a cumulative decrease in the EEM area and a greater lumen loss than segments with early ER (-2.5 +/- 3.4 vs -0.6 +/- 2.6 mm(2), p < 0.001). CONCLUSIONS: In transplant vasculopathy, the late remodeling response was different between segments with early constrictive and expansive remodeling, despite similar intimal thickening. Early constriction caused an overall decrease in EEM area and greater loss of lumen during follow-up. PMID- 12742414 TI - Prognostic value of contractile response during high-dose dipyridamole echocardiography test in heart transplant recipients. AB - BACKGROUND: Coronary allograft vasculopathy (CAV) remains a main factor limiting long-term survival after heart transplantation (HTX). The diagnosis of CAV is still based on serial coronary angiography. In this study, we evaluated the prognostic value of high-dose dipyridamole echocardiography in HTX. METHODS: Sixty-eight patients underwent dipyridamole echocardiography within 48 hours of their scheduled annual coronary angiography. Coronary allograft vasculopathy was defined as CAV 1 (focal or diffuse stenosis <50%) or CAV 2 (focal or diffuse stenosis >or=50%). Wall-motion score index (WMSI) was evaluated at rest and after dipyridamole administration. RESULTS: Results of coronary angiography were normal in 43 patients (63%), showed CAV 1 in 11 (16%), and showed CAV 2 in 14 (21%). Rest wall motion was normal in 39 patients and abnormal in 29. After dipyridamole administration, wall motion remained normal in all 39 (Group 1, no CAV in 34 and CAV 1 in 5). Of 29 patients with rest wall-motion abnormalities, all reversed to normal after dipyridamole in 8 patients (Group 2, no CAV in 7 and CAV 1 in 1) and remained or worsened in 21 (Group 3, CAV 2 in 14 and no CAV or CAV 1 in 7). During follow-up (6 +/- 3 years), 15 patients had major cardiac events: 11 occurred in Group 3, whereas 4 occurred in Groups 1 and 2. Wall motion at rest and after dipyridamole administration and CAV were independent predictors for cardiac events; only dipyridamole WMSI >1 remained significant (p < 0.0001) at multivariate analysis. CONCLUSIONS: Dipyridamole echocardiography is a simple, non-invasive test that after HTX may identify patients with altered wall motion who deserve stricter surveillance. PMID- 12742415 TI - Rapamycin inhibits vascular remodeling in an experimental model of allograft vasculopathy and attenuates associated changes in fibrosis-associated gene expression. AB - BACKGROUND: Rapamycin inhibits extracellular matrix (ECM) accumulation (fibrosis) and vascular remodeling in experimental models of chronic allograft dysfunction (CAD) by poorly understood mechanisms. The aim of this study was to assess the effect of rapamycin on the expression of fibrosis-associated genes and correlate this with observed changes in ECM remodeling in an experimental of model allograft vasculopathy. METHODS: Vascular remodeling and ECM accumulation (picrosirius red) were measured by computerized histomorphometry of F344-to-Lewis rat aortic allograft sections harvested at serial timepoints. Expression of fibrosis associated genes was studied by means of semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Rapamycin (0.5 mg/kg/day) inhibited intimal hyperplasia, medial ECM accumulation and expansive vascular remodeling (increasing vessel circumference) in rat aortic allografts. This was associated with attenuation of the graft inflammatory infiltrate and a reduction in intragraft gelatinase, collagen III and tissue inhibitor of metalloproteinase 1 (TIMP 1) mRNA levels. At a lower dosage (0.25 mg/kg/day), rapamycin inhibited intimal hyperplasia and medial ECM accumulation, but there was a lesser effect on vascular remodeling. Lower dose allografts were also seen to have a more severe inflammatory infiltrate and larger amounts of intragraft matrix metalloproteinase 9 (MMP 9) mRNA than those treated with the higher dose. CONCLUSIONS: These data suggest that, in addition to the tissue response to injury, the alloimmune injury itself may contribute directly to the vascular remodeling that occurs in allograft vasculopathy. Rapamycin at higher but not lower doses inhibited both of these pathologic processes. PMID- 12742416 TI - Mechanical support for the failing cardiac allograft: a single-center experience. AB - BACKGROUND: Mechanical support for pre-transplant stabilization is established, but its use in peri-operative graft failure (PGF) has not been well documented. With liberal acceptance criteria being used to enlarge the donor pool, an increased incidence of graft failure might be expected. We evaluated the incidence and outcome of PGF at our institution. METHODS: A retrospective review of 462 consecutive adult heart transplants performed between January 1993 and December 1999 revealed 20 cases of PGF. Donor-, surgery- and device-related variables were evaluated for association with operative mortality, survival and successful device weaning. RESULTS: Transplant recipients included 17 men and 3 women, median age 56.5 years (20 to 66 years). PGF etiology included primary graft failure (n = 9); right heart failure (RHF) secondary to pulmonary hypertension, coagulopathy/intra-operative hemorrhage or sepsis (n = 9); and hyperacute rejection (n = 2). Device types included RVAD (n = 11), LVAD (n = 4), BIVAD (n = 3) and IABP (n = 2). The wean rate was 45%. Duration of device support ranged from 2 to 965 hours. Early ventricular recovery (within 96 hours) was associated with significantly better 30-day and 2-year survival. Weaned patients had an 88% 30-day and 67% 2-year survival, whereas the overall survival rate was 79% at 2 years (p = not significant). CONCLUSIONS: Early ventricular recovery is an important predictor of successful weaning and survival. In view of the prohibitive mortality associated with PGF and the dismal prognosis with re transplantation, we advocate aggressive use of mechanical assistance for PGF, with an acceptable survival benefit. PMID- 12742417 TI - Spectrum of left ventricular dysfunction in potential pediatric heart transplant donors. AB - BACKGROUND: Although myocardial dysfunction severe enough to preclude harvest for transplantation has been reported in approximately 20% of potential adult organ donors, the incidence of myocardial dysfunction in the pediatric population has not been studied systematically. Therefore, the purpose of this study was to determine the spectrum of myocardial dysfunction in potential pediatric heart transplant donors. METHODS: We reviewed the pediatric cardiology database at Primary Children's Medical Center to identify all children who had screening echocardiograms for potential organ donation. We reviewed charts for patient age and size, cause of brain death, and type of pharmacologic support. Echocardiograms were reviewed retrospectively for left ventricular systolic function (shortening or ejection fraction), wall motion abnormalities, diastolic function (mitral E/A ratio), Tei index, and mitral regurgitation. RESULTS: We identified 23 potential donors (age, 6.7 +/- 4.4 years; range, 5 days to 15 years). All patients were receiving pharmacologic support. We found systolic left ventricular dysfunction, defined as an ejection fraction <50% or a shortening fraction <28%, in 57% (13/23) of the patients. We found mitral regurgitation in 85% (11/13) of the patients with systolic dysfunction and in zero of 10 with normal ejection phase indices. Diastolic dysfunction (mitral E/A reversal) was found in 45% (6/13) of those with systolic dysfunction and in 60% (6/10) of patients with normal systolic ejection phase indices. The Tei index was abnormal in 8 of 13 patients with left ventricular systolic dysfunction (range, 0.5-1.4), and was normal in the 6 patients with isolated diastolic dysfunction. One patient, who was electrocuted, had regional wall motion abnormalities. Of the 23 potential donors, 19 (87%) had evidence of systolic or diastolic dysfunction. A total of 13 hearts (3 with normal systolic function, 4 with systolic dysfunction, and 6 with isolated diastolic dysfunction) were harvested for transplantation. CONCLUSIONS: Left ventricular systolic and diastolic dysfunctions are common findings in potential pediatric organ donors. Despite this, previous studies have shown that some of these hearts can be transplanted successfully. We speculate that some of the abnormalities occur as a physiologic consequence of brain death and, thus, may be reversible after transplantation. To avoid wasting a valuable, limited resource, further study is needed to identify the donors suitable for pediatric heart transplantation. PMID- 12742418 TI - Reduction of tricuspid annular doppler tissue velocities in pediatric heart transplant patients. AB - BACKGROUND: Orthotopic heart transplantation is a life-saving therapy for children with end-stage heart disease. However, 50% of these transplanted children die or require re-transplantation 12 years later. Progressive deterioration of cardiac function is a common feature of long-term survivors; however, quantitative evaluation of the state of the right ventricle has been lacking. Tissue Doppler imaging (TDI) has been used to measure alterations in right ventricular (RV) function in other illnesses. The purpose of this study was to quantitate abnormalities in tricuspid annular systolic and diastolic velocities as an indicator of RV dysfunction, and to evaluate if time since transplantation and the presence of tricuspid regurgitation are associated with quantitative changes in tricuspid annular velocities in pediatric heart transplant recipients. METHODS: TDI was performed and velocities recorded during systole and early and late diastole at the tricuspid annulus, septum and mitral annulus in transplanted patients and in a control group with normal hearts. Pulsed wave Doppler mitral and tricuspid inflows were also measured and the severity of tricuspid regurgitation was estimated using color flow mapping. Patients with biopsy evidence of active cellular rejection or left ventricular ejection fraction of <60% were excluded from study. RESULTS: Thirty-five patients were divided into a normal heart group (n = 14) and a transplant group (n = 21), aged from 1 to 23 years. Systolic and early diastolic velocities at the tricuspid annulus and septum in the transplant group were reduced significantly compared with the normal group (p < 0.05): tricuspid annular systolic, 5.8 +/- 1.4 vs 9.4 +/- 1.7 cm/sec; early diastolic, -6.4 +/- 2.6 vs -9.7 +/- 2.6 cm/sec; septum systolic, 3.9 +/- 1.5 vs 5.8 +/- 1.4 cm/sec; and early diastolic, -6.3 +/- 2.4 vs -9.1 +/- 2.5 cm/sec. Patients were divided into early (<5 years) and late (>5 years) term groups since transplantation. Tissue velocities at the tricuspid annulus in the late term group had further reduction in systole, 4.9 +/- 1.4 vs 6.4 +/- 1.1 cm/sec, and early diastole, -5.3 +/- 1.5 vs -7.1 +/- 2.9 cm/sec (p < 0.05). Patients with severe tricuspid regurgitation had systolic and early diastolic velocities at the tricuspid annulus that were further reduced. Left ventricular mitral inflow Doppler early/late diastolic ratios became significantly different from the normal group 5 years after transplantation (p < 0.05). CONCLUSIONS: TDI demonstrated that tricuspid annular systolic and early diastolic velocities were abnormal in children after transplantation and became significantly more abnormal with prolonged time after transplantation. These alterations were not dependent on the presence of severe tricuspid regurgitation but appeared to be exacerbated by its presence. Evidence of diastolic left ventricular dysfunction was not detected before 5 years after transplantation in this unselected group. A prospective study may be required to define the evolution and progression of right and left ventricular dysfunction in children after heart transplantation. PMID- 12742420 TI - Donor-transmitted coronary atherosclerosis. AB - BACKGROUND: Autopsies show that coronary atherosclerosis is present frequently in the young and healthy. However, according to our former guideline, we performed pre-transplant evaluation without coronary angiogram in donors <60 years. The purpose of this study is to evaluate to what extent native coronary atherosclerosis is transmitted through heart transplantation. METHODS: Between April 1986 and December 2000, a total of 1253 patients underwent heart transplantation at our institution. If coronary evaluation with coronary angiogram or autopsy had been performed within 6 months after transplantation, we regarded focal and non-circumferential atherosclerosis with >or=50% stenosis in proximal segments of at least 1 coronary vessel of the donor heart as transmitted, native coronary atherosclerosis, rather than newly developed transplant vasculopathy. RESULTS: We excluded 85 of 1253 (6.8%) cases because coronary evaluation was not performed within 6 months (n = 45) or because hearts underwent angiography during pre-transplant evaluation (n = 40). Of these, 2 patients with significant coronary atherosclerosis underwent transplantation and concurrent coronary artery bypass grafting. The prevalence of significant (stenosis >or=50%) and inadvertently transmitted coronary atherosclerosis was 7.0% (82/1168). CONCLUSION: The prevalence of coronary atherosclerosis in patients who underwent angiography within 6 months after transplantation was 5.2% (49/950). Among subjects who had autopsies within the first 6 months after heart transplantation, we found significant coronary atherosclerosis (stenosis >or=50%) 15.1% (33/218), and among those with early graft failure (<10 days after transplantation), the prevalence was 22.8% (26/114). The prevalence of coronary atherosclerosis in the donor pool is high, and donor screening without coronary angiogram overlooks significant coronary atherosclerotic lesions (stenosis >or=50%) in a considerable number of cases (7.0%). Because donor-transmitted coronary atherosclerosis is a risk factor in short-term (early graft failure) survival after heart transplantation, we have now changed our policy to include coronary angiography as a standard in screening donors >or=40 years. However, to what extent donor coronary atherosclerosis is accepted undoubtedly must be made arbitrarily until an evidence-based algorithm becomes available. PMID- 12742419 TI - Immune and non-immune factors in cryopreserved tissues. AB - BACKGROUND: Although cryopreserved tissues are used clinically, the effects of cryopreservation on antigenicity leading to immune and non-immune responses are not well known. METHODS: We investigated the change of inflammatory effects of cryopreserved tissue by using spleen and aortic allografts from Class I antigen disparate B6.C-H-2(bm1) (bm1; K(bm1), IA(b), D(b)), Class II antigen-disparate B6.C-H-2(bm12) (bm12; K(b), IA(bm12), D(b)) and Class I and Class II antigen disparate (bm1 x bm12)F1 (K(bm1 x b), IA(b x bm12), D(b)) mice against C57BL/6 Cr Slc (B6; H-2(b)) mice. Cryopreservation was done in a programmed freezer and cryopreserved tissues were kept in the vapor phase of liquid nitrogen for 2 weeks and thawed at room temperature. RESULTS: Cryopreserved B6 spleen cells expressed almost the same levels of Class I (K(b) and D(b)) and Class II (IA(b)) antigens as observed in fresh B6 spleen cells. Cryopreserved bm1 and bm12 spleen cells had the same stimulator activities in mixed-lymphocyte reaction (MLR) and cytotoxic T lymphocyte (CTL) assays compared with fresh bm1 and bm12 spleen cells, respectively. To elucidate the effects of cryopreserved tissues on immune response of recipients, descending aortas of (bm1 x bm12)F1 mice were implanted into the right common carotid artery of B6 (H-2(b)) mice with the cuff technique and the reactivities of recipient B6 mice against Class I antigen-disparate bm1 antigens and Class II antigen-disparate bm12 antigens were examined 4 weeks after implantation. In both MLR and CTL assays against bm1 or bm12 antigens, anti-donor reactivities were augmented and there was no significant difference between B6 mice grafted with fresh aortic allografts and those grafted with cryopreserved ones. Histologic analysis showed that mild infiltration of mononuclear cells into the adventitia was observed in both fresh and cryopreserved aortic allografts. The fibrous change was observed more strongly in cryopreserved aortic allografts compared with fresh aortic allografts. CONCLUSIONS: Cryopreservation has no effect on eliciting immune responses to Class I or Class II alloantigens, but has some effect on promoting fibrous change. PMID- 12742421 TI - Using fructose-1,6-diphosphate during hypothermic rabbit-heart preservation: a high-energy phosphate study. AB - BACKGROUND: In this study, we evaluated the effects of fructose-1,6-diphosphate (FDP) on high-energy phosphate metabolism during 18-hour hypothermic rabbit-heart preservation. METHODS: Under general anesthesia and artificial ventilation, hearts from 42 adult New Zealand white rabbits were harvested, flushed, and preserved in St. Thomas solution at 4(o)C for 18 hours. In the study group (n = 15), FDP (5 mmol/liter) was added to the St. Thomas solution, whereas in the control group (n = 17), fructose (5 mmol/liter) was added. Another 10 hearts did not undergo hypothermic storage, but were used as the normal group for high energy phosphate concentration comparison. RESULTS: After 18 hours of hypothermic preservation, myocardial high-energy phosphate content decreased in both preservation groups. In the study group, left ventricular adenosine triphosphate (ATP) content was 33% of that in the normal hearts, but in the control group, ATP decreased to 14% of normal. Adenosine diphosphate (ADP) content, energy charge, and ATP-to-ADP ratio showed similar decreases. The high-energy phosphate profile (content in the atria and ventricles and the ratio of ATP to ADP to AMP) was maintained in the study group but not in the control group. High-energy phosphate metabolites such as inosine monophosphate (IMP), inosine, and hypoxanthine increased in both preservation groups, but the increase was more prominent in the control group. CONCLUSION: Adding FDP to St. Thomas solution attenuated the depletion of high-energy phosphate concentration in the preserved hearts. This difference was especially prominent in the left and right ventricles. The protective effect of FDP during hypothermic heart preservation deserves further study. PMID- 12742422 TI - Significance of early bronchoscopic airway abnormalities after lung transplantation. AB - Abnormal bronchoscopic allograft airway appearance with erythema, friability and sloughing-presumed to be ischemic-has been described early after lung transplantation, but its association with post-operative spirometry or other outcomes is largely unknown. We conducted a prospective cohort study to assess the impact of such airway changes in all lung transplant recipients from January 1997 to July 2000. Twenty-four (59%) of 41 patients were classified as having ischemic-appearing airways. Timing and value of peak pulmonary function tests, mortality, incidence of infection, rejection, anastomotic stricture and bronchiolitis obliterans syndrome were not significantly different when compared to patients with normal-appearing airways over a mean follow-up period of 761.2 +/- 412.4 days. PMID- 12742423 TI - Pharmacokinetics of mycophenolic acid associated with calcineurin inhibitors: long-term monitoring in stable lung recipients with and without cystic fibrosis. AB - Pharmacologic interactions and absorption disturbances after transplantation may induce serologic fluctuation of immunosuppression and adversely affect outcome. We present data showing that trough levels of mycophenolic acid decreased by 50% during combined mycophenolate mofetil (MMF) and cyclosporine therapy compared with levels during combined MMF and tacrolimus therapy. In addition, cystic fibrosis patients required 30% higher doses of MMF to achieve the therapeutic levels of recipients without cystic fibrosis. PMID- 12742424 TI - Thymoglobuline use in pediatric heart transplantation. AB - The literature has few data regarding the use of polyclonal anti-thymocyte globulin in pediatric cardiac transplantation. We describe our single-center, retrospective study of the use of Thymoglobuline in a pediatric population. We included in the study 31 consecutive heart transplant recipients (mean age, 7.8 years; median age, 9 years; range, 4 months-17 years), who all survived surgery. To induce immunosuppression, all patients received Thymoglobuline therapy at age dependent doses (1-1.5 mg/kg/day between 0 and 1 year; 1.5-2 mg/kg/day from 1 year to 8 years; and 2.5 mg/kg/day >8 years). Duration of treatment was 1 to 7 days. In patients <1 year, the total number of lymphocytes was maintained at >500/mm(3). Thirty of 31 patients are alive at the end of follow-up. During the first 3 months, 3 Grade 3A and 10 Grade 1A (Working Formulation grading system) rejection episodes occurred. All reversed after steroid treatment. Eleven viral infections, 2 bacterial infections, and 1 fungal infection occurred. Not all patients with infection were symptomatic but all responded successfully to treatment. One episode of post-transplantation lymphoproliferative disease regressed after decreasing immunosuppression therapy and after acyclovir therapy. At the end of follow-up, 19 patients are without steroids. Immunosuppression therapy with Thymoglobuline is safe in the pediatric age group if the number of lymphocytes is monitored strictly. PMID- 12742425 TI - Total left ventricular outflow tract obstruction due to left ventricular assist device-induced sub-aortic thrombosis in 2 patients with aortic valve bioprosthesis. AB - This report describes 2 patients with an aortic bioprosthesis. Both patients developed total thrombotic occlusion of the sub-aortic left ventricular outflow tract consequent to insertion of a left ventricular assist device (LVAD). Replacing a mechanical valve with a bioprosthesis in patients receiving a left ventricular assist device offers no additional protection against thrombosis of the aortic prosthesis. Pericardial patching below the aortic prosthesis at the time of LVAD implantation may be performed, but will significantly impede or prohibit the native ventricle from ejecting blood and demonstrating any degree of recovery. PMID- 12742426 TI - Transcatheter closure of residual atrial septal defect after cardiac transplantation. AB - A 55-year-old male patient experienced 2 acute neurologic events 3 weeks after orthotopic cardiac transplantation. Transesophageal echocardiography demonstrated a patent foramen ovale in the native portion of the interatrial septum with bidirectional shunting by Doppler and microbubble contrast. The defect was closed successfully with a CardioSeal transcatheter septal closure device. This case demonstrates the advantages of the percutaneous approach for closure of residual defects in the post-operative patient. PMID- 12742427 TI - Fulminant mixed humoral and cellular rejection in a cardiac transplant recipient: a review of the histologic findings and literature. AB - Mixed acute cellular and humoral rejection is diagnosed uncommonly among heart transplant recipients and usually occurs within the first post-transplant month. We report a case of fatal, fulminant, mixed, acute cellular and humoral rejection in a 33-year-old woman 6 weeks after orthotopic heart transplantation. She had been treated with intravenous methylprednisolone for International Society for Heart and Lung Transplantation (ISHLT) Grade 2 rejection at post-operative Day 28. Despite intensification of immunosuppression therapy, she developed fever and progressive hemodynamic instability. Autopsy results revealed ISHLT Grade 4 mixed cellular and humoral rejection. Cellular rejection is a well-described mechanism of graft failure early after heart transplantation. Although humoral rejection also is recognized as contributing to early graft failure, its characteristics and clinical implications are not as well characterized. We describe a patient with fulminant mixed rejection, despite intensified immunosuppression therapy, early after orthotopic heart transplantation who presented with high-grade fever. We include a review of the literature on humoral and mixed rejections. PMID- 12742428 TI - Effects of water and water-free polar solvents on the tensile properties of demineralized dentin. AB - OBJECTIVES: The aim of this study was to test the null hypothesis that the tensile properties of demineralized dentin are not influenced by the hydrogen bonding ability of anhydrous polar solvents. METHODS: Dentin disks 0.5mm thick were prepared from mid-coronal dentin of extracted, unerupted, human third molars. 'I' beam and hour-glass shaped specimens were prepared from the disks, the ends protected with nail varnish and the central regions completely demineralized in 0.5M EDTA for 5 days. Ultimate tensile stress (UTS) and low strain apparent modulus of elasticity (E) were determined with the specimens immersed for 60 min in water, methanol, HEMA, acetone or air prior to testing in those same media. Apparent moduli of elasticity were measured on the same specimens in a repeated measures experimental design. The results were analyzed with a one-way ANOVA on ranks, followed by Dunn's test at alpha=0.05. Regression analysis examined the relationship between UTS or E and Hansen's solubility parameter for hydrogen bonding (delta(h)) of each solvent. RESULTS: The UTS of demineralized dentin in water, methanol, HEMA, acetone and air was 18(7), 29(7), 31(6), 41(13) and 146(27)MPa, x(SD), n=10. Low-strain E for the same media were 11(7), 43(12), 79(21), 132(31) and 253(115)MPa. Regression analysis of UTS vs delta(h) revealed a significant (p<0.0005, r=-0.69, R(2)=0.48) inverse, exponential relationship. A similar inverse relationship was obtained between low strain E vs delta(h) (p<0.0001, r=-0.93, R(2)=0.86). SIGNIFICANCE: The tensile properties of demineralized dentin are dependent on the hydrogen bonding ability of polar solvents (delta(h)). Solvents with low delta(h) values may permit new interpeptide H-bonding in collagen that increases its tensile properties. Solvents with high delta(h) values prevent the development of these new interpeptide H-bonds. PMID- 12742429 TI - Effect of hydrolyzed surface layer on the cytotoxicity and chemical resistance of a low fusing porcelain. AB - OBJECTIVE: The objectives of this study were to verify the formation of a hydrolyzed surface layer on Duceram LFC and to determine its effect on the cytotoxicity of the porcelain as measured by cellular activity and concentrations of leached ionic species. METHODS: Specimens were fabricated from dentin porcelain by a vibration blotting technique. Half of the specimens underwent accelerated aging by subjecting them to the standard test for hydrolytic resistance (ISO 6872). Fibroblast cell cultures were placed in direct contact with specimens. Cell viability was assessed using succinic dehydrogenase activity. Chemical resistance was determined by leaching specimens with a continuous flow of deionized water and measuring the concentrations of soluble ions in the leachates. Porcelain surface topography was examined using atomic force microscopy, and Fourier transform infrared spectroscopy was used to detect the composition of the surface layer. RESULTS: Hydrolyzation treatment created a smooth texture on the porcelain surfaces but did not result in a hydrolyzed surface layer with increased hydroxyl content. There was a decreased alkali ion content in the surface layer of hydrolyzed specimens. Mean cellular SDH activities for non-hydrolyzed and hydrolyzed porcelain were 75+/-7 and 80+/-5% of Teflon controls, respectively. Only sodium ions were present in significant concentrations in collected leachates. The sodium concentration decreased over the initial 4.5h of leaching. SIGNIFICANCE: The changes in surface layer texture and composition as the result of the aging treatment had little effect on the cytotoxicity and chemical resistance of Duceram LFC. The results suggest that Duceram LFC would pose no biocompatibility risk even after extended exposure to the oral environment. These data also provide baseline material properties to be used in future studies of the effects of porcelain additives. PMID- 12742430 TI - Effects of different whiskers on the reinforcement of dental resin composites. AB - OBJECTIVE: Whiskers were recently used to reinforce dental composites to extend their use to large stress-bearing restorations. The aim of this study was to investigate the effects of different types of whiskers on composite properties. METHODS: Silicon nitride and silicon carbide whiskers were each mixed with silica particles at whisker/silica mass ratios of 0:1, 1:5, 1:2, 1:1, 2:1, 5:1, and 1:0, and thermally treated. The composite was heat-cured at 140 degrees C. Strength and fracture toughness were measured in flexure, while elastic modulus and hardness were measured with nano-indentation. RESULTS: Both whisker type and whisker/silica ratio had significant effects on composite properties (two-way ANOVA; p<0.001). Silicon nitride whiskers increased the composite strength and toughness more than did silicon carbide. Silicon carbide whiskers increased the modulus and hardness more than silicon nitride did. The silicon nitride whisker composite reached a strength (mean+/-SD; n=6) of 246+/-33 MPa at whisker/silica of 1:1, while the silicon carbide whisker composite reached 210+/-14 MPa at 5:1. Both were significantly higher than 114+/-18 MPa of a prosthetic control and 109+/-23 MPa of an inlay/onlay control (Tukey's multiple comparison test; family confidence coefficient=0.95). Fracture toughness and work-of-fracture were also increased by a factor of two. Higher whisker/silica ratio reduced the composite brittleness to 1/3 that of the inlay/onlay control. SIGNIFICANCE: Whisker type and whisker/silica ratio are key microstructural parameters that determine the composite properties. Reinforcement with silica-fused whiskers results in novel dental composites that possess substantially higher strength and fracture toughness, and lower brittleness than the non-whisker control composites. PMID- 12742431 TI - Detection of in vitro demineralization adjacent to restorations using quantitative light induced fluorescence (QLF). AB - AIM: Quantitative light-induced fluorescence (QLF) is a technique for the detection, quantification, and longitudinal monitoring of early carious lesions. The technique is non-destructive and can be used in vivo. Using the natural fluorescence of teeth, and the loss of such fluorescence in demineralized enamel, QLF is a repeatable and valid optical caries monitor. Previously used in smooth and occlusal surfaces, the purpose of this pilot study was to determine if QLF could detect, and longitudinally monitor, demineralization adjacent to a range of restorative materials. METHODS: Fifteen previously extracted lower third molars were selected based upon the lack of any visible demineralization. A single burr hole was placed on the buccal surface and the cavity restored with amalgam, composite, compomer, glass ionomer or a temporary filling material. The buccal surface was then coated in an acid resistant nail varnish leaving an exposed area around the restoration and also a similar sized control region. The teeth had QLF images taken at baseline and were then subjected to a demineralizing buffer, further QLF images were subsequently taken at 72 and 144 h. Transverse microradiography was used to confirm the presence of early, subsurface lesions at the completion of the cycle (144 h). QLF images were analyzed by a single blinded examiner and values for change in radiance fluorescence were computed. These values were recorded as loss of radiance fluorescence loss integrated over area of lesion and expressed as DeltaQ. RESULTS: The appearance of each material under QLF and the change in fluorescence is described. Amalgam, glass ionomer and the temporary material all exhibited reduced fluorescence, while composite and compomer showed increased fluorescence, when compared with surrounding enamel. There was no change in fluorescence of the materials when subjected to experimental demineralizing conditions. Readings at 72 and 144 h demonstrated demineralization adjacent to the restorations and at the exposed control. Significant differences were detected between baseline, 72 and 144 h using ANOVA on all restorations with the exception of compomer where significance was noted between baseline and 144 h, p>0.05. CONCLUSIONS: This pilot study has demonstrated the ability for QLF to detect and monitor secondary caries. Analysis techniques should be based upon the subtraction of baseline DeltaQ scores from subsequent images. Further research is required to assess the ability of QLF to detect secondary lesions in vivo. PMID- 12742432 TI - Grindability of cast Ti-Cu alloys. AB - OBJECTIVE: The purpose of the present study was to evaluate the grindability of a series of cast Ti-Cu alloys in order to develop a titanium alloy with better grindability than commercially pure titanium (CP Ti), which is considered to be one of the most difficult metals to machine. METHODS: Experimental Ti-Cu alloys (0.5, 1.0, 2.0, 5.0, and 10.0 mass% Cu) were made in an argon-arc melting furnace. Each alloy was cast into a magnesia mold using a centrifugal casting machine. Cast alloy slabs (3.5 mm x 8.5 mm x 30.5 mm), from which the hardened surface layer (250 microm) was removed, were ground using a SiC abrasive wheel on an electric handpiece at four circumferential speeds (500, 750, 1000, or 1250 m/min) at 0.98 N (100 gf). Grindability was evaluated by measuring the amount of metal volume removed after grinding for 1min. Data were compared to those for CP Ti and Ti-6Al-4V. RESULTS: For all speeds, Ti-10% Cu alloy exhibited the highest grindability. For the Ti-Cu alloys with a Cu content of 2% or less, the highest grindability corresponded to an intermediate speed. It was observed that the grindability increased with an increase in the Cu concentration compared to CP Ti, particularly for the 5 or 10% Cu alloys at a circumferential speed of 1000 m/min or above. SIGNIFICANCE: By alloying with copper, the cast titanium exhibited better grindability at high speed. The continuous precipitation of Ti(2)Cu among the alpha-matrix grains made this material less ductile and facilitated more effective grinding because small broken segments more readily formed. PMID- 12742433 TI - Cytotoxic effects of packable and nonpackable dental composites. AB - OBJECTIVE: Advanced dental packable and nonpackable composite materials have been recently introduced onto the market as 'amalgam alternatives' for the restoration of posterior teeth. Most established composites are applied in increments of no more than 2mm and light cured to ensure complete polymerization, whereas new formulations have been claimed to be suitable for application in increments of 4 5mm. The aims of the present study were to analyze the cytotoxicity of these new composites in comparison to an established nonpackable composite using standardized cell culture systems and to examine the influence of thickness of the light cured increments on the cytotoxicity of these materials. METHODS: Specimens were prepared in polyethylene tubes covered with mylar. All materials were light cured in 2.5 or 5mm increments with a Demetron curing light (light intensity: 550 mW/cm(2)). Specimens were added to the cultures immediately after production or after preincubation for 1, 2, 7 days or 6 weeks under cell-culture conditions. Specimens were incubated with L-929 fibroblasts for 72 h and cell numbers determined by flow cytometry. In a different series of experiments, dopaminergic cells were incubated with composite supernatants. RESULTS: Results with L-929 fibroblasts demonstrated that all freshly prepared composite materials reduced cell numbers (p<0.05) in comparison to controls. The cytotoxicity of all substances diminished after increasing preincubation times (p<0.0001). In a rank order of cytotoxicity, the established composite and two of the advanced composite materials exhibited least cytotoxicity, whereas the other advanced composites showed moderate or severe cytotoxic effects. The cytotoxicity of each material was much higher when polymerized in 5mm increments than in 2.5mm composite increments (p<0.0001). SIGNIFICANCE: Our data demonstrate that the advanced composites tested show similar or more severe cytotoxicity than an established nonpackable composite and that cytotoxicity of all materials investigated increases when applied in a 5mm bulk increment. PMID- 12742434 TI - In vitro characterization of two laboratory-processed resin composites. AB - PURPOSE: To compare various characteristics of two new-generation laboratory processed resin composites (BelleGlass HP/SDS-Kerr and Sinfony/3M-ESPE). The properties evaluated were degree of C=C conversion, microhardness, roughness, biaxial flexural strength and polymerization shrinkage-strain. MATERIALS AND METHODS: All specimens were subjected to a first and a second polymerization cycle according to the manufacturers' instructions. The degree of C=C conversion (DC) was recorded on rectangular (3 x 2 x 0.5mm(3)) specimens (n=3) by FT-IR micromultiple internal reflectance spectroscopy immediately after each of the two polymerization cycles. Twenty cylindrical specimens (10 x 2mm(2)) of each material were prepared for surface microhardness (n=10, VHN, 200 g load, 20s) and surface roughness (n=10, Ra) measurements. The biaxial flexural strength and stiffness were determined on disk-shaped (n=8, 15 x 0.7 mm(2)) specimens loaded to fracture at 1 mm/min crosshead speed. The polymerization shrinkage-strain was calculated with the bonded-disk method. All values were statistically analyzed by Student's unpaired t-test (p<0.05). RESULTS: The second polymerization cycle significantly increased the degree of C=C conversion for both materials (p<0.05). BelleGlass HP exhibited significantly higher degree of C=C conversion, surface microhardness, surface roughness, biaxial flexural strength and stiffness values compared to Sinfony (p<0.05). SIGNIFICANCE: Several differences exist between the materials although both products are recommended for the same clinical applications. PMID- 12742435 TI - Degradation of resin-dentin bonds using NaOCl storage. AB - OBJECTIVE: Several recent studies have reported collagen hydrolysis within bonds over the long-term. This may be one reason for the degradation of the bonds. This study therefore aimed to determine the effects of NaOCl on adhesive system bonds (total-etch bonding system vs. self-etching primer system) to dentin in order to accelerate the durability testing. METHODS: Resin-dentin bonded specimens were produced using Liner Bond 2V (Kuraray), a self-etching primer system, and OptiBond SOLO (Kerr), a total-etch bonding system, according to the manufacturers' instructions. The bonded specimens were serially sectioned in both x and y directions across the adhesive interface to obtain beams (adhesive area: 0.9 mm(2)). The specimens were immersed in 10% NaOCl solution for 1-5h after being stored in water at 37 degrees C for 24h. Control specimens were tested without exposure to NaOCl. After storage, micro-tensile bond tests were performed. Results were analyzed by two-way ANOVA and Fisher's PLSD tests (p<0.05). All fractured surfaces were observed by SEM, and examined using an image analyzer. RESULTS: The bond strengths decreased with increasing storage time in NaOCl. Fractography showed that NaOCl had a greater effect on the bond structure of OptiBond SOLO than on that of Liner Bond 2V, although both adhesives were susceptible. SIGNIFICANCE: Deterioration of the bonds was responsible for the effect of NaOCl on the hybrid layer. This deterioration may occur in humans in cases of deproteinization within the bonds. PMID- 12742436 TI - Irradiance effects on the mechanical properties of universal hybrid and flowable hybrid resin composites. AB - OBJECTIVES: A potential problem with high-intensity lights might be failure of polymer chains to grow and cross-link in a desired fashion, thereby affecting the structure and properties of the polymers formed. The purpose of this study was to evaluate mechanical properties of resin composites polymerized using four different light-curing units. METHODS: A conventional quartz-tungsten-halogen (QTH) light, a soft-start light, an argon-ion laser, and a plasma-arc curing light were used to polymerize disk-shaped (9.0mm diameter x 1.0 mm high) and cylinder-shaped (4mm diameter x 8 mm high) specimens of a universal hybrid and a flowable hybrid composite. Biaxial flexure strength, fracture toughness, hardness, compressive strength, and diametral tensile strength were determined for each composite. RESULTS: The use of the plasma-arc curing light, a high intensity light, resulted in significantly lower hardness for the universal hybrid composite compared with the hardness obtained using the conventional QTH and the soft-start units. Hardness was the only mechanical property that was adversely affected by the use of a high-intensity light. SIGNIFICANCE: High intensity lights might affect some resin composite mechanical properties, but this effect cannot be generalized to all resin composites and all properties. PMID- 12742437 TI - Aspects of water sorption from the air, water and artificial saliva in resin composite restorative materials. AB - OBJECTIVE: Primarily to establish whether artificial saliva (AS) at 37 degrees C is essential as a clinically relevant environment for testing filled, resin composite restorative materials. The effect of other storage conditions was also investigated for comparison and controls: desiccation, exposure to the laboratory atmosphere, high humidity cabinet, saturated water vapor, and deionized water. METHODS: Two visible light-cured products were used: Heliomolar Radiopaque (HR) and Tetric Ceram (TC) (Ivoclar, Schaan, Liechtenstein). Bar specimens (26 x 1.5 x 1.0 mm(3)) were cured at five overlapping spots for 60s per spot and randomly distributed into groups of six. Trial 1: one group of each material was exposed first to atmospheric air at 24 degrees C, approximately 50% RH (24WV(50)), then to water vapor at 37 degrees C, approximately 97% RH (37WV(97)), and then immersed in deionized water at 37 degrees C (37DW). Trial 2 used three groups of each material, one first exposed to 37WV(97) followed by 37DW, the other two were immediately immersed in 37DW or artificial saliva (37AS). Trial 3: two groups of each material were vacuum desiccated at 37 degrees C, then exposed to 37 degrees C, approximately 100% RH (37WV(100)), then immersed in 37DW or 37AS. Trial 4: four groups of HR were treated similarly to Trial 3; one was left under desiccation, and another in 37WV(100) for the remaining period. Three-point bend tests for flexural strength (F), flexural modulus (E), and total energy to failure (W) were performed at the end of Trials 2-4. RESULTS: Environmental moisture absorption was substantial at 24WV(50)(c. 0.2%), at least 40% of that in 37DW (HR: c. 0.7%, TC: c. 0.5%). Saturation was achievable in 37WV(100). Mass loss on desiccation (HR: c. 0.4-0.5%, TC: c. 0.25%) was reversible in 37WV(100). There were some significant effects of exposure conditions on mechanical properties (e.g. F for HR: after desiccation, 85.7+/-1.4MPa; after 37WV(100), 73.2+/-3.6MPa; difference: p<0.0002), but overall the results were unclear. After a rapid gain in mass, there was a gradual loss in both 37DW and 37AS for both materials, slightly more in 37AS than 37DW. SIGNIFICANCE: Water vapor absorption is substantial, hence attention must be paid to the laboratory working environment and conditions of storage and testing, i.e. temperature and RH must be stated to assist interpretation of data and comparisons between studies. Test conditions need to be standardized and with reference to normal oral conditions, immediate immersion in artificial saliva at 37 degrees C is the preferred treatment for these materials, whatever time of testing is chosen. PMID- 12742438 TI - A 6-year clinical evaluation of Class I poly-acid modified resin composite/resin composite laminate restorations cured with a two-step curing technique. AB - OBJECTIVE: Polymerization shrinkage is still one of the main disadvantages of resin composite restorations (RC). Especially in cavities with a high C-factor, debonding can occur. A laminate restoration including a base with a more elastic behavior might result in a better adaptation. The purpose of this study was to evaluate the durability of a combination of two techniques suggested to counter the stress formation in direct RC restorations in cavities with the highest C factor. METHODS: Each of the 29 patients received one or two pair(s) of Class I restorations. The first restoration was a poly-acid modified resin composite/resin composite (PMRC/RC) sandwich restoration and the second a direct RC restoration. Both restorations, except for the PMRC layer, were placed with oblique layering and two-step curing technique. Ninety restorations, 23 premolar and 67 molars, were evaluated annually with slightly modified USPHS criteria during 6 years. RESULTS: At 6 years, 41 pairs were evaluated. A cumulative failure rate of 2.4% was observed for both the RC and the laminate restorations. One laminate restoration showed non-acceptable color match, but was not replaced and one RC restoration showed non-acceptable marginal adaptation. Two cases of slight postoperative sensitivity were observed in one patient. Three restorations were partially replaced due to primary proximal caries. SIGNIFICANCE: A high durability for and no differences, were observed between both restorative techniques in Class I cavities. PMID- 12742439 TI - Effect of artificial saliva contamination on pH value change and dentin bond strength. AB - OBJECTIVE: This study was to examine the effect of artificial saliva contamination on pH change of the dentin surface and the micro-shear bond strength (MSBS) of the two bonding systems to contaminated dentin. METHODS: Fifty six human dentin disks were tested with two resin bonding systems: a self-etching primer system, Clearfil SE Bond (Kuraray Medical Inc., Tokyo, Japan), and a one bottle adhesive system, Single Bond (3M-ESPE, St. Paul, MN). Dentin surfaces were conditioned with the self-etching primer (primer) or phosphoric acid (etchant) and divided into four groups: conditioning without contamination (conditioning), contamination with artificial saliva (contamination), re-conditioning the contaminated dentin (re-conditioning), water-rinsing the contaminated dentin and re-conditioning (rinsing and re-conditioning). The pH change on the dentin surface was measured using a pH-imaging microscope (SCHEM-100, Horiba Ltd, Kyoto, Japan) to estimate the acid-base characteristics of the conditioned and contaminated dentin surface. The MSBS to the dentin was examined after storage in distilled water at 37 degrees C for 1 week. RESULTS: The pH of intact dentin surfaces was 6.9. Conditioning with the primer and etchant decreased the pH to 5.4 and 5.9, respectively. Saliva contamination increased the pH slightly, and re conditioning decreased the pH again. The MSBS of the two bonding systems decreased after contamination. Re-priming restored the MSBS to control values, while re-etching did not. Rinsing and re-priming produced a reduction in MSBS, and rinsing and re-etching did not increase the MSBS. SIGNIFICANCE: The conditioning and saliva contamination changed the pH value of dentin surface. The MSBS decreased after contamination; however, re-priming with SE Bond primer was an adequate treatment to restore the bond strength. PMID- 12742440 TI - Flexural fatigue behavior of resin composite dental restoratives. AB - OBJECTIVE: The aim of this study was to evaluate the mechanical properties of resin composite dental restoratives under quasi-static and cyclic loading. METHODS: Four-point-bending bars of 10 different resin composite materials were manufactured according to ISO standard and stored for two weeks in distilled water. The fracture strength (FS) was measured with the four-point-bending test in an universal testing machine. The flexural fatigue limits (FFL) for 10(5) cycles were determined under equivalent loading. All specimens were tested and fatigued in water at 37 degrees C. The data were analyzed using ANOVA, Weibull statistics of FS and the 'staircase' approach of FFL. Fractographic analysis was performed using SEM. RESULTS: The initial flexural strength values for the resin composite materials varied from 55.4 MPa for Solitaire up to 105.2 MPa for Filtek Z250. The mean flexural fatigue limit for 10(5) cycles ranged between 37 and 67% of the initial strength. SEM analysis of the fractured surfaces suggests two kinds of failure mechanisms for initial and fatigue fracture. SIGNIFICANCE: The fatigue behavior of resin composite materials does not correlate with initial strength values. Materials providing high initial strengths do not obviously reveal the best fatigue resistance. Flexural fatigue measurement of resin composite materials should be viewed as a useful tool to evaluate long term mechanical properties. PMID- 12742441 TI - Silane treatment effects on glass/resin interfacial shear strengths. AB - OBJECTIVE: Methacrylic resin-based dental composites normally use a bifunctional silane coupling agent with an intermediary carbon connecting segment to provide the interfacial phase that holds together the organic polymer matrix with the reinforcing inorganic phase. In this study, fiber pull-out tests were used to measure the interfacial bond strength at the fiber-matrix interface. METHODS: Glass fibers (approximately 30 microm diameter, 8 x 10 (-2)m length, MoSci) were silanated using various concentrations (1, 5 and 10%) of either 3 methacryloxypropyl-trimethoxysilane (MPS) or glycidoxypropyltrimethoxy-silane (GPS) in acetone (99.8%). Rubber (poly(butadiene/acrylonitrile), amine terminated, M(w) 5500) molecules were also attached to the fiber surface via GPS molecules. The resin was comprised of a 60/40 mixture of Bis-phenol-A bis-(2 hydroxypropyl)-methacrylate (BisGMA) and tri (ethylene glycol) dimethacrylate (TEGDMA). A bead of resin approximately 2-4 x 10(-3)m in embedded length was placed on the treated fibers and light cured. The load required to pull the fiber out of the resin was converted to shear bond strength. RESULTS: Interfacial shear strengths were greater for silanated specimens compared with unsilanated, and for MPS compared with GPS. The same set of samples soaked in 50:50 (v/v) mixtures of ethanol and distilled water for a period of 1 month showed a decrease in properties. SIGNIFICANCE: A positive correlation was found between the amount of silane on the filler surface and the property loss after soaking. Rubber treatment provided improvement in interfacial strength. 5% MPS samples had the highest strength both in soaked as well as unsoaked samples. PMID- 12742442 TI - Measuring morphological parameters of the pelvic floor for finite element modelling purposes. AB - The goal of this study was to obtain a complete data set needed for studying the complex biomechanical behaviour of the pelvic floor muscles using a computer model based on the finite element (FE) theory. The model should be able to predict the effect of surgical interventions and give insight into the function of pelvic floor muscles. Because there was a lack of any information concerning morphological parameters of the pelvic floor muscle structures, we performed an experimental measurement to uncover those morphological parameters. Geometric parameters as well as muscle parameters of the pelvic floor muscles were measured on an embalmed female cadaver. A three-dimensional (3D) geometric data set of the pelvic floor including muscle fibre directions was obtained using a palpator device. A 3D surface model based on the experimental data, needed for mathematical modelling of the pelvic floor, was created. For all parts of the diaphragma pelvis, the optimal muscle fibre length was determined by laser diffraction measurements of the sarcomere length. In addition, other muscle parameters such as physiological cross-sectional area and total muscle fibre length were determined. Apart from these measurements we obtained a data set of the pelvic floor structures based on nuclear magnetic resonance imaging (MRI) on the same cadaver specimen. The purpose of this experiment was to discover the relationship between the MRI morphology and geometrical parameters obtained from the previous measurements. The produced data set is not only important for biomechanical modelling of the pelvic floor muscles, but it also describes the geometry of muscle fibres and is useful for functional analysis of the pelvic floor in general. By the use of many reference landmarks all these morphologic data concerning fibre directions and optimal fibre length can be morphed to the geometrical data based on segmentation from MRI scans. These data can be directly used as an input for building a mathematical model based on FE theory. PMID- 12742443 TI - Postural stability of the human mandible during locomotion. AB - Movements of the head and of the mandible relative to the head were measured in human subjects walking and running on a treadmill at various speeds and inclinations. A miniature magnet and piezo-electric accelerometer assembly was mounted on the mandibular incisors, and a Hall-effect sensor along with a second accelerometer mounted on a maxillary incisor along a common vertical axis. Signals from these sensors provided continuous records of vertical head and mandible acceleration, and relative jaw position. Landing on the heel or on the toe in different forms of locomotion was followed by rapid deceleration of the downward movement of the head and slightly less rapid deceleration of the downward movement of the mandible, i.e., the mandible moved downwards relative to the maxilla, then upwards again to near its normal posture within 200 ms. No tooth contact occurred in any forms of gait at any inclination. The movement of the mandible relative to the maxilla depended on the nature and velocity of the locomotion and their effects on head deceleration. The least deceleration and hence mandibular displacement occurred during toe-landing, for example, during "uphill" running. The maximum displacement of the mandible relative to the head was less than 1mm, even at the fastest running speed. The mechanisms that limit the vertical movements of the jaw within such a narrow range are not known, but are likely to include passive soft-tissue visco-elasticity and stretch reflexes in the jaw-closing muscles. PMID- 12742444 TI - An EMG-driven musculoskeletal model to estimate muscle forces and knee joint moments in vivo. AB - This paper examined if an electromyography (EMG) driven musculoskeletal model of the human knee could be used to predict knee moments, calculated using inverse dynamics, across a varied range of dynamic contractile conditions. Muscle-tendon lengths and moment arms of 13 muscles crossing the knee joint were determined from joint kinematics using a three-dimensional anatomical model of the lower limb. Muscle activation was determined using a second-order discrete non-linear model using rectified and low-pass filtered EMG as input. A modified Hill-type muscle model was used to calculate individual muscle forces using activation and muscle tendon lengths as inputs. The model was calibrated to six individuals by altering a set of physiologically based parameters using mathematical optimisation to match the net flexion/extension (FE) muscle moment with those measured by inverse dynamics. The model was calibrated for each subject using 5 different tasks, including passive and active FE in an isokinetic dynamometer, running, and cutting manoeuvres recorded using three-dimensional motion analysis. Once calibrated, the model was used to predict the FE moments, estimated via inverse dynamics, from over 200 isokinetic dynamometer, running and sidestepping tasks. The inverse dynamics joint moments were predicted with an average R(2) of 0.91 and mean residual error of approximately 12 Nm. A re-calibration of only the EMG-to-activation parameters revealed FE moments prediction across weeks of similar accuracy. Changing the muscle model to one that is more physiologically correct produced better predictions. The modelling method presented represents a good way to estimate in vivo muscle forces during movement tasks. PMID- 12742445 TI - The primary stability of a cementless stem varies between subjects as much as between activities. AB - The rehabilitation program adopted immediately after a cementless total hip replacement is a very important factor, because of the known relationship between osseointegration and implant micromotion. The present study was aimed to evaluate which type of task is the most critical in terms of bone-implant relative micromotion. Both inter-task and inter-subject variability were taken into account to verify if the movement strategy could be determinant on this assessment. A previously validated finite element model was used to predict the peak total micromovements over the entire bone-implant contact surface in four different patients, performing nine different tasks, using published data on joint forces recorded by instrumented hip prostheses. The results predicted by the various simulations suggest that while stair climbing is surely a critical task for primary stability, for some subjects other tasks may be as critical as stair climbing. From a variance analysis for simple crossover design on the predicted peak micromotion, the inter-subject variability had much more influence on the primary stability of cementless implant than the inter-task variability. Even if the results of Patient IBL, who was reported to have difficulties to perform any activities in a normal way, were excluded from the statistical analysis, the inter-subject variability remained still higher than the inter-task variability. The results obtained from simulations suggest that the strategy the hip replacement patient adopts to perform a given motor task, may be, for the implant stability, equally or even more critical than the type of motor task performed. PMID- 12742446 TI - Modelling heat transfer in a bone-cement-prosthesis system. AB - The heat transfer in a general bone-cement-prosthesis system was modelled. A quantitative understanding of the heat transfer and the polymerization kinetics in the system is necessary because injury of the bone tissue and the mechanical properties of the cement have been suggested to be effected by the thermal and chemical history of the system. The mathematical model of the heat transfer was based on first principles from polymerization kinetics and heat transfer, rather than certain in vitro observed properties, which has been the common approach. Our model was valid for general three-dimensional geometries and an arbitrary bone cement consisting of an initiator and monomer. The model was simulated for a cross-section of a hip with a potential femoral stem prosthesis and for a cement similar to Palacos R. The simulations were conducted by using the finite element method. These simulations showed that this general model described an auto accelerating heat production and a residual monomer concentration, which are two phenomena suggested to cause bone tissue damage and effect the mechanical properties of the cement. PMID- 12742447 TI - Cancellous and cortical morselized allograft in revision total hip replacement: A biomechanical study of implant stability. AB - To restore femoral intramedullary bone stock loss in revision surgery of failed total hip arthroplasties, impacted morselized cancellous allograft is recommended. This study investigated the mechanical properties of both impacted cortical (group A) and cancellous (group B) morselized bone graft for reconstruction of femoral bones. Ten matched pairs of fresh frozen human femora were prepared by over-reaming to create a smooth-walled cortical shell. Each pair had one cortical and one cancellous impacted morselized allograft and cement. Stem subsidence was evaluated by a cyclic axial load, which was applied by a servohydraulic test. The stem subsidence was measured for initial subsidence (subsidence at the first 1000 cycles), the total axial subsidence (subsidence at the end of cycles under load) and the final axial subsidence (subsidence after the unloading phase). Torque test was measured by torsional loads through the prosthetic femoral heads. Total axial subsidence was significantly higher in group B (mean: 1.32+/-0.32 mm) compared to group A (mean: 0.94+/-0.26 mm) (P<0.01). There was no significant difference between the two groups in terms of initial subsidence (P=0.09) and final axial subsidence. The mean maximum torque before failure was 39.5+/-22.2 N-m for the cortical morselized allograft and 32.5+/-18.1N-m for cancellous. We concluded that impacted morselized cortical bone graft used for reconstruction of contained femoral bone loss in revision hip arthroplasty, may reduce the stem subsidence. Further animal experimentation for mechanical and histological evaluation of in vivo application is warranted. PMID- 12742448 TI - Non-driving intersegmental knee moments in cycling computed using a model that includes three-dimensional kinematics of the shank/foot and the effect of simplifying assumptions. AB - Assessing the importance of non-driving intersegmental knee moments (i.e. varus/valgus and internal/external axial moments) on over-use knee injuries in cycling requires the use of a three-dimensional (3-D) model to compute these loads. The objectives of this study were: (1) to develop a complete, 3-D model of the lower limb to calculate the 3-D knee loads during pedaling for a sample of the competitive cycling population, and (2) to examine the effects of simplifying assumptions on the calculations of the non-driving knee moments. The non-driving knee moments were computed using a complete 3-D model that allowed three rotational degrees of freedom at the knee joint, included the 3-D inertial loads of the shank/foot, and computed knee loads in a shank-fixed coordinate system. All input data, which included the 3-D segment kinematics and the six pedal load components, were collected from the right limb of 15 competitive cyclists while pedaling at 225 W and 90 rpm. On average, the peak varus and internal axial moments of 7.8 and 1.5 N m respectively occurred during the power stroke whereas the peak valgus and external axial moments of 8.1 and 2.5 N m respectively occurred during the recovery stroke. However, the non-driving knee moments were highly variable between subjects; the coefficients of variability in the peak values ranged from 38.7% to 72.6%. When it was assumed that the inertial loads of the shank/foot for motion out of the sagittal plane were zero, the root-mean squared difference (RMSD) in the non-driving knee moments relative to those for the complete model was 12% of the peak varus/valgus moment and 25% of the peak axial moment. When it was also assumed that the knee joint was revolute with the flexion/extension axis perpendicular to the sagittal plane, the RMSD increased to 24% of the peak varus/valgus moment and 204% of the peak axial moment. Thus, the 3-D orientation of the shank segment has a major affect on the computation of the non-driving knee moments, while the inertial contributions to these loads for motions out of the sagittal plane are less important. PMID- 12742449 TI - The mechanics of back-extensor torque production about the lumbar spine. AB - The purpose of this study was to develop and evaluate a biomechanical model of lumbar back extension over a wide range of positions for the lumbar spine, incorporating the latest information on muscle geometry and intra-abdominal pressure (IAP). Analysis of the Visible Human data was utilised in order to obtain anatomical information unavailable from the literature and magnetic resonance imaging was used to generate subject-specific anatomical descriptions. The model was evaluated by comparisons with measured maximal voluntary static back-extension torques. Predicted maximal specific muscle tensions agreed well with in vitro measurements from the literature. When modelling the maximal static back-extension torque production, it was possible to come fairly close to simultaneous equilibrium about all the lumbar discs simply by a uniform muscle activation of all back-extensor muscles (the caudal part showed, however, less agreement). This indicates that equilibrium in the lumbar spine is mainly regulated by passive mechanical properties, e.g. muscle length changes due to postural changes, rather than due to complex muscle coordination, as earlier proposed. The model showed that IAP (measured during torque exertions) contributes about 10% of the total maximal voluntary back-extensor torque and that it can unload the spine from compression. The spinal unloading effect from the IAP was greatest with the spine held in a flexed position. This is in opposition to the effects of changed muscle lever arm lengths, which for a given load would give the largest spinal unloading in the extended position. These findings have implications for the evaluation of optimal lifting techniques. PMID- 12742450 TI - Explicit calibration method and specific device designed for stereoradiography. AB - The three-dimensional geometry of the human spine is noteworthy information that can be obtained by stereoradiographic methods. These methods are based on the identification of anatomical structures in several views which are obtained by rotation of a patient standing on a turntable. Calibration algorithms for computer vision or photogrammetry are well documented, but they generally yield calibration devices which are cumbersome for the use in clinical stereoradiography. This paper presents a calibration method adapted to a two-view stereoradiography calibration (frontal and lateral incidences) and based on a simplified geometric modeling of the radiological environment. The a priori knowledge yields four calibration equations related to the vertical and horizontal planes of both views, leading to a specific calibration procedure and device. Moreover this device is attached to the stereoradiographic system (directly integrated on the turntable) in order to facilitate clinical applications. A validation was performed on 26 dried lumbar vertebrae in order to evaluate clinical situation. The mean accuracy of the stereoradiographic reconstruction was 1.2mm. PMID- 12742451 TI - In vitro performance of intramedullary cement restrictors in total hip arthroplasty. AB - Contemporary cementing techniques in total hip arthroplasty include the use of a cement restrictor to occlude the intramedullary canal. As there are many different designs currently available it was the aim of our study to compare the stability of eight different systems. We investigated the displacement and the ability to occlude the femur of these cement restrictors during standardised cementing of artificial and fresh frozen femora. The maximal intramedullary pressures and the displacement of the plugs were continuously recorded and statistically evaluated. The results revealed significant differences between the tested cement restrictors. The expandable REX Cement Stop and the Exeter Plug achieved the highest stability and the least cement leakage. The more rigid designs (Palacos Plug, BUCK, Universal) in contrast showed inferior performance. Our biomechanical study emphasises the importance of cement restrictor selection, which can have a crucial influence on the fixation of a cemented total hip replacement. PMID- 12742452 TI - Pathways of load-induced cartilage damage causing cartilage degeneration in the knee after meniscectomy. AB - Results of both clinical and animal studies show that meniscectomy often leads to osteoarthritic degenerative changes in articular cartilage. It is generally assumed that this process of cartilage degeneration is due to changes in mechanical loading after meniscectomy. It is, however, not known why and where this cartilage degeneration starts. Load induced cartilage damage is characterized as either type (1)--damage without disruption of the underlying bone or calcified cartilage layer--or type (2), subchondral fracture with or without damage to the overlying cartilage. We asked the question whether cartilage degeneration after meniscectomy is likely to be initiated by type (1) and/or type (2) cartilage damage. To investigate that we applied an axisymmetric biphasic finite element analysis model of the knee joint. In this model the articular cartilage layers of the tibial and the femoral condyles, the meniscus and the bone underlying the articular cartilage of the tibia plateau were included. The model was validated with data from clinical studies, in which the effects of meniscectomy on contact areas and pressures were measured. It was found that both the maximal values and the distributions of the shear stress in the articular cartilage changed after meniscectomy, and that these changes could lead to both type (1) and type (2) cartilage damage. Hence it likely that the cartilage degeneration seen after meniscectomy is initiated by both type (1) and type (2) cartilage damage. PMID- 12742453 TI - Increasing strain and strain rate strengthen transient stiffness but weaken the response to subsequent compression for articular cartilage in unconfined compression. AB - Strain amplitude and strain rate dependent nonlinear behavior and load-induced mechanical property alterations of full-thickness bovine articular cartilage attached to bone were investigated in unconfined compression. A sequence of test compressions of finite deformation (ranging from 0.9% to 34.5% nominal strain) was performed at strain rates ranging from approximately 0.053%/s to 5.8%/s. Peak and equilibrium loads were analyzed to determine strain amplitude and strain rate dependence of linear versus nonlinear responses. The test protocol was designed to reveal changes in mechanical properties due to these finite deformations by interspersing small-amplitude witness ramps of approximately 1.1% deformation and approximately 0.44%/s strain rate between the test ramps ("witness" meaning to assess any mechanical property changes). We found that peak loads displayed high nonlinearity, stiffening with both increasing compression amplitude and more so with increasing strain rate. The response to witness ramps suggested that mechanical weakening occurred when compression amplitude reached 1.9-2.9% strain and beyond, and that weakening was much more significant at higher strain rate. These findings delineate regimes of linear versus nonlinear behavior of cartilage, and indicate the types of loads which can cause mechanical property alterations. Biological implications of this study are that strain amplitude and strain rate dependent stiffening may be essential to bear physiological loads and to protect cells and matrix from mechanical damage. Structural changes reflected by mechanical weakening at small compression could also initiate remodeling or disease processes. PMID- 12742454 TI - Prevention of mesh-dependent damage growth in finite element simulations of crack formation in acrylic bone cement. AB - Peak stress levels predicted in finite element analysis (FEA) usually depend on mesh density, due to singular points in the model. In an earlier study, an FEA algorithm was developed to simulate the damage accumulation process in the cement mantle around total hip replacement (THR) implants. It allows cement crack formation to be predicted, as a function of the local cement stress levels. As the simulation is driven by mesh-dependent peak stresses, predicted crack formation rates are also likely to be mesh dependent. The aim of this study was to evaluate the mesh dependence of the predicted crack formation process, and to present a method to reduce the mesh dependence. Crack-propagation experiments were simulated. Experimental specimens, representing transverse slices of cemented THR reconstructions, were subjected to cyclic torsional loading. Crack development around the corners of the stem was monitored. The experiments were simulated using three meshes with increasing levels of mesh refinement. Crack locations and orientations were accurately predicted, and were virtually independent of the level of mesh refinement. However, the experimental crack propagation rates were overestimated considerably, increasing with mesh refinement. To eliminate the effect of stress singularities around the corners of the stem, a stress averaging algorithm was applied in the simulation. This algorithm redistributed the stresses by weighted spatial averaging. When damage accumulation was computed based on averaged stresses, the crack propagation rates predicted were independent of the level of mesh refinement. The critical distance, a parameter governing the effect of the averaging algorithm, was optimized such that the predicted crack propagation rates accurately corresponded to the experimental ones. These results are important for the validity and standardization of pre-clinical testing methods for orthopaedic implants. PMID- 12742455 TI - A new model-based RSA method validated using CAD models and models from reversed engineering. AB - Roentgen stereophotogrammetric analysis (RSA) was developed to measure micromotion of an orthopaedic implant with respect to its surrounding bone. A disadvantage of conventional RSA is that it requires the implant to be marked with tantalum beads. This disadvantage can potentially be resolved with model based RSA, whereby a 3D model of the implant is used for matching with the actual images and the assessment of position and rotation of the implant. In this study, a model-based RSA algorithm is presented and validated in phantom experiments. To investigate the influence of the accuracy of the implant models that were used for model-based RSA, we studied both computer aided design (CAD) models as well as models obtained by means of reversed engineering (RE) of the actual implant. The results demonstrate that the RE models provide more accurate results than the CAD models. If these RE models are derived from the very same implant, it is possible to achieve a maximum standard deviation of the error in the migration calculation of 0.06 mm for translations in x- and y-direction and 0.14 mm for the out of plane z-direction, respectively. For rotations about the y-axis, the standard deviation was about 0.1 degrees and for rotations about the x- and z axis 0.05 degrees. Studies with clinical RSA-radiographs must prove that these results can also be reached in a clinical setting, making model-based RSA a possible alternative for marker-based RSA. PMID- 12742456 TI - Representation of passive spinal element contributions to in vitro flexion extension using a polynomial model: illustration using the porcine lumbar spine. AB - A polynomial modeling approach was developed to describe the contribution of individual passive spinal elements to the lumbar spinal motion segment flexion extension motion. Flexion-extension moment-angle curves from porcine lumbar spines tested using a robotic testing system were described using sixth-order polynomials; the polynomials describing different dissection conditions were subtracted to describe the contribution of individual spinal elements to the motion segment flexion-extension properties. This modeling approach is a powerful and straightforward method for representing the mechanics of individual spinal tissues in biomechanical models and could easily be expanded to incorporate other features such as axial load. PMID- 12742457 TI - Slide track analysis of the relative motion between femoral head and acetabular cup in walking and in hip simulators. PMID- 12742462 TI - Sirolimus: its discovery, biological properties, and mechanism of action. AB - Sirolimus is the USAN-assigned generic name for the natural product rapamycin. Sirolimus is produced by a strain of Streptomyces hygroscopicus, isolated from a soil sample collected from Rapa Nui commonly known as Easter Island. Although sirolimus was isolated as an antifungal agent with potent anticandida activity, subsequent studies revealed impressive antitumor and immunosuppressive activities. Sirolimus demonstrates activity against several murine tumors, such as B16 43 melanocarcinoma, Colon 26 tumor, EM ependymoblastoma, and mammary and colon 38 solid tumors. Sirolimus is a potent inhibitor of antigen-induced proliferation of T cells, B cells, and antibody production. Demonstration of the potent immunosuppressive activity of sirolimus in animal models of organ transplantation led to clinical trials and subsequent approval by regulatory authorities for prophylaxis of renal graft rejection. Interest in sirolimus as an immunosuppressive therapy in organ transplantation derives from its unique mechanism of action, its unique side-effect profile, and its ability to synergize with other immunosuppressive agents. The molecular mechanism underlying the antifungal, antiproliferative, and immunosuppressive activities of sirolimus is the same. Sirolimus forms an immunosuppressive complex with intracellular protein, FKBP12. This complex blocks the activation of the cell-cycle-specific kinase, TOR. The downstream events that follow the inactivation of TOR result in the blockage of cell-cycle progression at the juncture of G1 and S phase. PMID- 12742463 TI - The development of immunosuppression: the rapamycin milestone. PMID- 12742464 TI - The Rapamune era of immunosuppression 2003: the journey from the laboratory to clinical transplantation. AB - The story of Rapamune (sirolimus, rapamycin) began with the isolation of an antibiotic from a soil sample sent to Ayerst Laboratories in Montreal. More than 25 years later, sirolimus was approved for use by transplant physicians in the United States. Development programs for new drugs for transplantation face significant challenges. Four key challenges were critical to the development of sirolimus as a drug for transplantation: First, sirolimus was not intended to be an antirejection agent. Second, sirolimus was not easy to make or purify into a palatable substance for human use and the development of a pharmaceutical form was an important and critical hurdle. Third, sirolimus showed potent antirejection activity when tested in de novo allograft recipients, but the development program required careful attention to its optimal use in multidrug transplant regimens. Fourth, the clinical program approved in the United States was rejected in Europe, and it was only with additional studies and a unique appeal process that sirolimus became available in Europe. Currently, sirolimus (Rapamune) is available throughout most of the world except in Japan, having achieved regulatory approvals in North America, Europe, the Middle East, Latin America, and Asia. Although sirolimus failed in its original role as an antifungal agent, it ultimately succeeded as an antirejection drug. Today, sirolimus holds additional promise both as a drug useful for the prevention of restenosis after coronary angioplasty and as an antitumor agent. PMID- 12742465 TI - Ten years of sirolimus therapy for human renal transplantation: the University of Texas at Houston experience. PMID- 12742466 TI - Two-year results of multicenter phase III trials on the effect of the addition of sirolimus to cyclosporine-based immunosuppressive regimens in renal transplantation. AB - Randomized clinical trials conducted over 24 months and including nearly 1300 renal transplant patients compared the efficacy and safety of two dose levels of sirolimus versus azathioprine (United States) or placebo (Global) comparators administered with a cyclosporine (CsA) and prednisone (Pred) baseline regimen. Analysis of 24-month data revealed that patients in the 5 mg/d sirolimus groups experienced a significant delay in the onset and reduction in the incidence of acute rejection episodes compared with azathioprine (Aza) or placebo groups (P =.02/P =.001). Graft and patient survival rates and also the occurrence of transplant-related infections, lymphoproliferative disorders, or malignancies were similar among all treatment arms. Between 12 and 24 months, patients treated with 2 mg/d sirolimus displayed relatively stable mean serum creatinine values, namely 1.9-1.8 mg/dL for the US and 1.8-1.8 mg/dL for the Global (P = NS) studies, which remained higher than those of the comparators. Both 5 mg/d groups showed an increase in mean serum creatinine during this interval, which was significantly higher than the value in both comparators at 24 months. Both sirolimus groups showed persistently elevated triglyceride levels compared with Aza-treated patients at month 24. The Global trial showed a less-pronounced difference in mean fasting triglyceride values compared with placebo. Data from both trials demonstrate that the addition of sirolimus to a CsA-Pred treatment regimen yielded a durable immunosuppressive effect associated with a progressive resolution of adverse side effects over time except for hyperlipidemia, which required continued countermeasure therapy. PMID- 12742467 TI - Sirolimus therapy without calcineurin inhibitors: Necker Hospital 8-year experience. PMID- 12742468 TI - Bicetre hospital experience with sirolimus-based therapy in human renal transplantation: the Sirolimus European Renal Transplant Study. AB - In 11 European centers, first cadaveric renal allograft recipients were randomized to CsA (n = 42) or sirolimus (n = 41). Dosing of these agents was concentration-controlled and open-labeled. All patients received corticosteroids and azathioprine. At 12 months, graft survival (98% sirolimus vs 93% CsA), patient survival (100% vs 98%), and incidence of biopsy-confirmed acute rejection (41% vs 38%) were similar. Serum creatinine was lower with sirolimus, significantly (P 50% and recipients of regrafts who had lost their first graft from rejection within the first 6 months were ineligible. The groups were compared for graft function, incidence of rejection, and patient and graft survival at 6 months. There were no differences in demographics. There were 30% and 29% discontinuations in the rTAC and sTAC groups mainly due to adverse events in the first month. The 6-month patient and graft survival by intention-to-treat analysis was 94% and 91% for rTAC and 100% and 97% for sTAC (P = NS), respectively. Incidence and severity of biopsy-proven acute rejection was not different between the two groups, being 21% for rTAC and 19% for sTAC. The mean serum creatinine was 121 micromol/L and 148 micromol/L for rTAC and sTAC groups (P =.09), respectively. Glomerular filtration rate (GFR) was 68 mL/min and 62 mL/min (P =.23), respectively. Adverse events, infections, and antihypertensive and antilipidemic agent usage were similar. Of interest is that the overall incidence of thrombotic microangiopathy was 14%. These results support the safety and efficacy of SRL + TAC. Reduced TAC is associated with a trend toward improved renal function. PMID- 12742483 TI - Kidney transplantation without calcineurin inhibitors using sirolimus. AB - INTRODUCTION: With the introduction of new immunosuppressive medicines, it has become possible to determine the extent to which nephrotoxic medicines contribute to CAN. The aim of this study is to compare the safety and efficacy of calcineurin inhibitor (CI) free immunosuppression in a prospective, randomized trial comparing sirolimus-mycophenolate mofetil (MMF)-prednisone to tacrolimus- MMF-prednisone. METHODS: Patients are randomized at the time of transplant to receive either tacrolimus (target level 12 to 15 ng/mL in the first month) or sirolimus (target level 12 to 18 ng/mL in the first month). All patients also receive MMF (750 mg bid) and prednisone tapered to 10 mg/d by 3 months and thymoglobulin induction (1.5 mg/kg/d on days 0, 1, 2, 4 and 6). RESULTS: At this point we have 4-month follow-up in 85 patients. The acute rejection rate is 7.5% (3/40) in the tacrolimus group and 6.7% (3/45) in the sirolimus group. We have discontinued sirolimus in eight patients so far, with wound complications being the most common indication. Renal function appears to be better in the sirolimus group at 1 month after transplantation, but the difference is not statistically significant. CONCLUSIONS: While longer follow-up is needed, these results demonstrate that total avoidance of CI can be achieved with extremely low acute cellular rejection rates using sirolimus-based immunosuppression in combination with thymoglobulin, MMF, and prednisone. PMID- 12742485 TI - Conversion to sirolimus in solid organ transplantation: a single-center experience. AB - BACKGROUND: Calcineurin inhibitors are associated with adverse events, including nephrotoxicity and diabetes that might reduce the benefits of long-term graft survival. We report our experience in converting kidney (K), kidney-pancreas (KP), pancreas (P), and (L) recipients from a calcineurin inhibitor/mycophenolate mofetil (MMF)/prednisone dose-induced nephrotoxicity (K = 9, KP = 5, P = 1, L = 5), hemolytic uremic syndrome (HUS) (K = 7, KP = 5), chronic allograft nephropathy (K = 12, L = 1), and glucose intolerance (K = 9, KP = 6, P = 2, L = 2). METHODS: The conversion protocol consisted of an abrupt discontinuation of the calcineurin inhibitor with sirolimus (8-12 mg, PO loading dose) initiated 24 72 hours after stopping the calcineurin inhibitor. Sirolimus was titrated to target trough levels of 12-16 ng/mL. Daclizumab 2 mg/kg IV was given to all KP and P recipients on days 0 and 14 postconversion. RESULTS: Resolution of HUS occurred in 12 of 12 patients (100%) with a drop in serum creatinine from 3.3 +/- 1.5 to 1.8 +/- 0.9 mg/dL (P =.04). Sirolimus conversion due to nephrotoxicity, HUS, and chronic allograft nephropathy improved serum creatinine from 2.9 +/- 1.4 to 2.2 +/- 0.9 mg/dL (P =.01). Eleven of 19 patients (58%) resolved glucose intolerance. Two patients suffered rejection due to noncompliance. Increases in cholesterol (208 +/- 70 to 243 +/- 77 mg/dL, P <.05) and triglycerides (232 +/- 145 to 265 +/- 148 mg/dL, P = NS), and minimal reduction in platelet values (243 +/- 85 to 237 +/- 85, P = NS) occurred. CONCLUSIONS: These data suggest that a calcineurin inhibitor-free immunosuppressive regimen with sirolimus, mycophenolate mofetil, and steroids preserves graft function in patients with clinical indications warranting calcineurin inhibitor discontinuation. PMID- 12742484 TI - Sirolimus monotherapy following Campath-1H induction. AB - Sirolimus was used as a single agent for maintenance immunosuppression in a pilot trial of 29 primary kidney transplant patients using lymphocyte depletion with Campath-1H as an induction strategy. This allowed sirolimus to be analyzed (dose, blood level, and side effect profile) in the absence of steroid and calcineurin inhibitors. A sirolimus dose of 4 mg/day resulted in blood levels in the 8 to 9 ng/mL range. Of the 29 patients, 8 patients (28%) had rejection. The sirolimus levels were not significantly different in patients with or without rejection. The cardiovascular risk profile in terms of lipid profile and hypertension control was favorable. Increase in cholesterol and triglyceride levels at one month (not statistically significant) necessitated treatment in 60% of patients with decline in levels by 6 and 12 months. Management of hypertension was also favorable with the majority of patients (55%) being on one hypertensive medication. Sirolimus monotherapy was well tolerated on the whole. Wound healing, leukopenia, and anemia were not significant problems. In conclusion, monotherapy has been well tolerated with a favorable side effect profile. However, a rejection rate of 28% was noted. PMID- 12742486 TI - Clinical use of rapamycin in renal allograft recipients identifies its relevant toxicity profile and raises unsolved questions: a single-center experience. AB - Rapamycin is an immunosuppressive drug with a distinct and unique mode of action and a specific side effect profile. We report here briefly on our personal clinical experience using this immunosuppressive drug in different combinations and settings. Rapamycin is without any doubt an efficient drug capable of preventing acute allograft rejection in a variety of immunosuppressive combinations. It is also a very potent drug that is not devoid of serious side effects. Infectious complications as a result of strong inhibition of the immune system are a frequent cause of hospitalization with severe morbidity and even mortality. Fungal infections and pneumonia are among the most devastating complications. As clinical experience with rapamycin grows and the therapeutic window of the drug can be further narrowed, these infectious complications will improve. Wound healing problems and lymphocoeles form another frequent surgical dilemma and are related to the antiproliferative properties of rapamycin. Last, hyperlipidemia warrants the use of statins in the majority of rapamycin-treated patients and whether this unfavorable side effect will offset the theoretically beneficial cardiovascular effects of the drug remains to be determined in controlled trials with long-term follow-up. Finally, the specific antiproliferative properties of rapamycin and the fact that it exerts no nephrotoxicity make this drug an alternative for calcineurin inhibitors and could make it an ideal candidate for treating chronic allograft dysfunction. PMID- 12742487 TI - Sirolimus changes lipid concentrations and lipoprotein metabolism in kidney transplant recipients. AB - BACKGROUND: Sirolimus (Rapammune, rapamycin, RAPA) is a strong immunosuppressive agent that reduces kidney transplant rejection. Hyperlipidemia is a significant side effect of sirolimus treatment and often leads to vascular disease. We have studied the repeatability, reversibility, and dose dependence of the plasma lipid and apoprotein changing effects of sirolimus and attempted to determine the mechanism by which sirolimus induces hypertriglyceridemia in some kidney transplant recipients. METHODS: Six patients with renal allografts maintained on cyclosporine A and prednisone were selected on the basis of their previous hyperlipidemic response to short-term (14 days) sirolimus administration. For longer-term treatment, each patient was started on 10 mg/d sirolimus and continued as tolerated for 42 days to reinduce hyperlipidemia. Timed blood samples were analyzed for lipid, apoprotein, and sirolimus levels. RESULTS: During sirolimus administration, mean total plasma cholesterol increased from 214 to 322 mg/dL (+50%); low density lipoprotein-cholesterol levels changed in a similar pattern. Mean triglyceride level rose from 227 to 432 mg/dL (+95%). ApoB 100 concentration rose from 124 to 160 mg/dL (+28%). ApoC-III level increased from 28.9 to 55.5 mg/dL (+92%). These lipid and apoprotein changes were found to be repeatable, reversible, and dose dependent. [(13)C(4)]-palmitate metabolic studies in four patients with hypertriglyceridemia indicated that the free fatty acid pool was expanded by sirolimus treatment (mean = 42.3%). Incorporation of [(13)C(4)]-palmitate into triglycerides of very low density lipoprotien, intermediate density lipoprotein, low density lipoproteins was decreased 38.3%, 42.1%, and 38.4%, respectively, by sirolimus treatment of these patients. CONCLUSIONS: These results suggest that sirolimus alters the insulin signaling pathway so as to increase adipose tissue lipase activity, decrease lipoprotein lipase activity, or both, resulting in increased hepatic synthesis of triglyceride, increased secretion of VLDL, and increased hypertriglyceridemia. PMID- 12742489 TI - Tubular function in patients with hypokalemia induced by sirolimus after renal transplantation. PMID- 12742488 TI - Cardiovascular risk factors of sirolimus compared with cyclosporine: early experience from two randomized trials in renal transplantation. AB - INTRODUCTION: Renal transplant recipients are at a higher risk of cardiovascular events, including death. This paper examines cardiovascular risk factors in two phase II studies comparing cyclosporine (CsA) with sirolimus-based therapy. METHODS: In two phase II studies, patients (n = 161) were randomized at the time of transplantation to receive either sirolimus or CsA in triple-therapy regimens with either azathioprine (Study A) or mycophenolate mofetil (Study B), and corticosteroids. Sirolimus whole blood trough levels were targeted to 30 ng/mL for 2 months and 15 ng/mL thereafter. Pooled results of the two studies are reported. RESULTS: When patients receiving sirolimus were compared with those receiving CsA, peak cholesterol and trigylcerides at 2 months were markedly and significant higher with sirolimus therapy. The difference between groups decreased thereafter and was not significant from 12 through 24 months. Control of lipid parameters in sirolimus-treated patients was achieved by decreasing the target trough levels after 2 months and by using lipid-lowering agents. Sirolimus based therapy was associated with a lower incidence of treatment-emergent hypertension (47.5% vs 29.6%, P <.024). At 24 months, the calculated glomerular filtration rate was significantly better with sirolimus (51.3 vs 65.1 mL/min, P <.001). There were no significant differences in the incidences of diabetes or death due to cardiovascular events. CONCLUSION: Patients receiving sirolimus experience an initial increase in lipid levels, but these effects are manageable with the use of lipid-lowering agents. Hypertension was less frequent and renal function was improved with CsA-free, sirolimus-based therapy. Based on this early experience, overall cardiovascular risk does not appear to be increased with sirolimus-based compared with CsA-based therapy. PMID- 12742492 TI - Sirolimus therapy in cardiac transplantation. AB - Rapamycin powerfully inhibits the progression of antigen-activated T cells through the cell cycle. In animal heart transplantation models, rapamycin therapy has been associated with profound immunosuppressive effects on host humoral and cellular responses. In consequence, further studies have been conducted to evaluate the efficiency of rapamycin in preventing acute heart allograft rejection, treating refractory acute heart allograft rejection, inducing transplantation tolerance, and preventing and treating transplant coronary artery disease. The results of these studies indicated that rapamycin can effectively prevent acute graft rejection and inhibit refractory acute graft rejection in heart transplant recipients by exerting potent immunosuppressive and antiproliferative effects without adversely affecting renal function. This supports the use of rapamycin therapy in heart transplant recipients, especially in those with renal dysfunction, for whom treatment with calcineurin inhibitors is contraindicated. Rapamycin may also halt and even reverse the progression of cardiac allograft vasculopathy, which warrants further clinical trials in humans. Finally, rapamycin may be able to induce transplantation tolerance, thus making it one of the most promising modalities for improving the long-term survival of heart transplant recipients. PMID- 12742491 TI - Use of rapamycin-impregnated stents in coronary arteries. AB - FIM STUDY: We investigated the 2-year safety and efficacy of sirolimus-eluting stents. Thirty patients had a single 18-mm sirolimus-eluting coronary stent implanted. Twenty-eight patients underwent angiographic and intravascular ultrasound follow-up at 2 years. No death occurred during the study period. No patient developed in-stent restenosis. One patient had a 52% in-lesion stenosis that required repeated revascularization and another patient underwent target vessel revascularization. Neointimal hyperplasia volume was minimal at 2 years in both groups. This study demonstrates the 2-year safety and efficacy of sirolimus eluting stenting. The slow release formulation showed slight superiority over the fast-release formulation in preventing late lumen loss, which was minimal in both groups. RAVEL TRIAL: This-study was a randomized, double-blind study that included 238 patients at 19 medical centers (15 in Europe, 3 in Brazil, and 1 in Mexico). Patients were eligible for the study if they were between 18 and 85 years of age, and had been given a diagnosis of stable or unstable angina or silent ischemia. Additional eligibility criteria were presence of a single primary target lesion in a native coronary artery that was 2.5 to 3.5 mm in diameter and that could be covered by an 18-mm stent stenosis of 51% to 99% of the luminal diameter and a flow rate of grade 1 or higher according to the Thrombolysis in Myocardial Infarction. RESULTS: One hundred twenty patients were randomly assigned to receive the sirolimus-eluting stent, and 118 were assigned to receive the standard stent. At 6 months, the degree of neointimal proliferation, manifested as the mean (+/-SD) late luminal loss, was significantly lower in the sirolimus-stent group (-0.01 +/- 0.33 mm) than in the standard-stent group (0.80 +/- 0.53 mm, P <.001). None of the patients in the sirolimus-stent group, as compared with 26.6% of those in the standard-stent group, had restenosis of >/=50% of the luminal diameter (P <.001). There were no episodes of stent thrombosis. During a follow-up period of up to 1 year, the overall rate of major cardiac events was 5.8% in the sirolimus-stent group and 28.8% in the standard-stent group (P <.001). The difference was due entirely to the higher rate of revascularization of the target vessel in the standard-stent group. CONCLUSION: Patients with angina who received sirolimus-eluting stents for the treatment of single, primary lesions in native coronary arteries had no angiographic evidence of late luminal loss or in-stent restenosis at 6 months, no episodes of thrombosis, and a very low rate of cardiac events at 1 year. PMID- 12742490 TI - Therapeutic monitoring of sirolimus: its contribution to optimal prescription. AB - It is now common practice to measure immunosuppressive drugs in blood as a guide to therapy. The immunosuppressive drug sirolimus, recently approved for use following kidney transplantation, was developed in the context of this clinical approach. Throughout the early clinical studies, validated analytical techniques based on chromatographic techniques were used to measure the drug. After a brief period in which an immunoassay was available, routine measurements are again being performed by chromatographic assays. In this article the use of blood concentration measurements in the assessment of the early and pivotal clinical trials of the drug is documented. Then, the rationale for the routine monitoring of the drug in clinical practice, a regulatory requirement in some countries, is set out. It is concluded that the development of this compound has benefited from experience gained during the pharmacokinetic assessment of other immunosuppressive drugs. The pharmacokinetic data accumulated on sirolimus have been a key element in formulating guidelines on dosing with this drug, both when used in combination with cyclosporine and when used after cyclosporine withdrawal. PMID- 12742493 TI - Safety and efficacy of sirolimus in kidney transplant patients and in patients with coronary artery disease undergoing angioplasty. AB - We show the key results of our 4-year experience with sirolimus in kidney transplant patients and in nontransplanted patients undergoing coronary angioplasty. METHODS: Recipients of one-haplotype living-related kidney allografts were randomized to receive sirolimus (2 mg/d, n = 35) or azathioprine (2 mg/kg per day, n = 35). Recipients of fully mismatched living kidney allografts (n = 55) received sirolimus (2 mg/day). High-risk recipients of black ethnicity (n = 68) were randomized to target whole-blood trough sirolimus concentrations between 8 and 12 ng/mL or 15 to 20 ng/mL. All kidney transplant patients received cyclosporine and prednisone. Sirolimus/cyclosporine pharmacokinetic studies were performed in 40 patients receiving 2 mg (n = 20) or 5 mg (n = 20) of sirolimus 7 days after transplantation. In the coronary intervention study, 12 patients at high risk for in-stent restenosis received sirolimus for 28 days after angioplasty. RESULTS: The incidence of biopsy confirmed acute rejection was 11.4% in recipients of one-haplotype living-related kidney allografts, 16.4% in recipients of fully mismatched living kidney allografts, and 15% (8 to 12 ng/mL) and 4% (15 to 20 ng/mL) in high-risk recipients of black ethnicity. Cyclosporine exposure was higher after morning administration compared to evening administration. There were poor correlations between sirolimus and cyclosporine exposures. The 4-month follow-up angiography revealed no restenosis (stenosis diameter > 50%), a late loss of 0.56 +/- 0.40 mm, and a loss index of 0.33 +/- 0.30. The follow-up 3D-intravascular ultrasound restudy showed an in-stent relative volumetric obstruction of 9.9 +/- 5.5%. Sirolimus in highly effective in preventing kidney allograft acute rejection and in-stent coronary restenosis. PMID- 12742494 TI - De novo use of sirolimus in immunosuppression regimens in kidney and kidney pancreas transplantation at the University of California, San Francisco. AB - At UCSF, we have used sirolimus in several immunosuppression regimens and protocols, and this article will summarize our experience in four areas. The purpose of the first study was to assess the efficacy of a sirolimus-based, calcineurin inhibitor-free regimen for the first 3 months after transplantation. Patients were treated with a calcineurin inhibitor-free regimen that consisted of daclizumab, sirolimus, mycophenolate mofetil (MMF), and conventional corticosteroids. Of nine patients, one (case #7) had an acute rejection episode (type IA) at 2 months after transplantation, which was fully reversed with corticosteroids. The second study was a prospective trial of calcineurin inhibitor-free regimen in patients with severe delayed graft function (DGF) (requiring dialysis). The immunosuppression regimen consisted of daclizumab, sirolimus, MMF, and corticosteroids. This immunosuppressive regimen was effective in patients with DGF; however, it was effective only in non-African American (non AA) patients (AA had a significantly higher acute rejection rate at 1 year than non-AA, 63% vs 23%, P =.025). In some patients sirolimus was associated with a prolonged recovery from DGF. The addition of sirolimus to immunosuppressive agents provide the opportunity for safe steroid withdrawal (at day 5). We participated in a sirolimus-based, multicenter open-label trial of very early corticosteroid withdrawal. Primary renal transplant patients were enrolled in an immunosuppression regimen that consisted of basiliximab, sirolimus (target levels 8 to 15 ng/mL, 0 to 5 months, and 6 to 12 ng/mL, 6 to 12 months) and tacrolimus in a dose of 0.05 mg/kg BID (target levels 6 to 9 ng/mL). Two of 14 enrolled patients had an episode of acute rejection before steroids were withdrawn. No acute rejection episodes have occurred after steroids were withdrawn (6-month follow-up). The regimen of sirolimus and tacrolimus was well tolerated. Wound complications were not noted. Another important use of sirolimus has been its incorporation in the immunosuppressive regimens in kidney-pancreas transplantation. Our current protocol consists of thymoglobulin induction, combined with MMF, sirolimus, and low-dose tacrolimus, for maintenance therapy. Steroids are only utilized during the first 5 to 6 days following the transplant. This steroid-free maintenance regimen has been used in the last 30 enteric drained, simultaneous pancreas-kidney transplants. Using this immunosuppressive approach, rejection rates are less than 10% for either the kidney or the pancreas. PMID- 12742495 TI - Sirolimus and islet transplants. PMID- 12742497 TI - Rapamycin in combination with cyclosporine or tacrolimus in liver, pancreas, and kidney transplantation. AB - A 10-year experience with the immunosuppressive drug rapamycin that begins in the laboratory then extends through multicentre trials in combination with cyclosporine in kidney transplant recipients, exploration of its use as a single agent and in combination with tacrolimus, and its potential in nonrenal organs is described. Rapamycin is a potent inhibitor of endothelial injury in rat aortic allografts. When added to full-dose cyclosporine it achieves low rejection rates, but it augments the nephrotoxicity and hyperlipidemia of cyclosporine. On the other hand, it allows discontinuation of calcineurin inhibitors in stable kidney and liver patients suffering from nephrotoxicity late posttransplant. At least in Caucasian patients, discontinuation of cyclosporine is possible as early as 3 months post-kidney transplant. In combination with low-dose tacrolimus, exceptionally low rates of rejection were seen in recipients of kidney, pancreas, and liver recipients with preservation of excellent renal function. These pilot studies have been confirmed in several single-centre and, more recently, multicentre trials in kidney and pancreas transplantation. The side-effect profile of hyperlipidemia, lymphocoeles, delayed wound healing, and possible liver effects are coming into focus, and ways of minimizing these problems being introduced. The lessons learned include the need for early adequate blood levels, the lack of correlation between dose and drug exposure, and the potency that allows marked dose reductions in calcineurin inhibitors and steroids. PMID- 12742496 TI - Sirolimus in liver transplantation. AB - Since its introduction in renal transplantation in 1999, sirolimus is being used by an increasing number of liver transplant centers. Compared to the calcineurin inhibitors, sirolimus acts through a separate signal transduction pathway and has a myriad of important biologic effects including: inhibition of lymphocyte proliferation, inhibition of fibrosis and fibroblast proliferation, and antineoplastic effects. The clinical side-effect profile of this drug is also different than calcineurin inhibitors. Most important, sirolimus does not cause glucose intolerance, hypertension, or renal insufficiency. As a result, this drug offers significant potential advantages over conventional immunosuppressive agents. However, sirolimus may cause hyperlipidemia and has also been associated with hepatic artery thrombosis in liver transplant recipients. This review will summarize the published data on sirolimus in liver transplantation, focusing on the potential advantages and disadvantage of the use of this drug in liver transplant recipients. Finally, the potential benefits of antifibrosis and antineoplastic effects of sirolimus in liver transplant recipients will be discussed. PMID- 12742498 TI - Ten years of sirolimus therapy in orthotopic liver transplant recipients. AB - BACKGROUND: Sirolimus therapy has been used in orthotopic liver transplant (OLT) recipients diagnosed with a variety of diseases; chronic graft rejection (CR), calcineurin associated renal toxicity, preemptive immune suppression, calcineurin related neurotoxicity, preemptive therapy in transplant recipients with history of hepatocellular carcinoma, and steroid resistant allograft rejection. METHODS: A search for the medical literature and experiences involving sirolimus was done. RESULTS: Several animal and human reports evaluating the use sirolimus in liver transplant recipients are found and discussed. CONCLUSION: Sirolimus has been used for multitude of indications, primarily based on anecdotal experiences. However, reports of sirolimus related side effects have decreased the transplant communities' enthusiasm towards promoting this agent as a safe immune suppression agent. PMID- 12742499 TI - Preclinical results of sirolimus treatment in transplant models. AB - Sirolimus (SRL; rapamycin) is a macrolide antibiotic, which modest anticandidal and tumoricidal activities were superseded by its immunosuppressive potential to block allograft rejection. The most intriguing biological characteristic of SRL emerged after demonstration of its potent synergism with cyclosporine (CsA). Naive T cells, residing in the G(0) phase of the cell cycle, become activated by three signals. Signal 1 (T cell antigen receptor/alloantigen) and Signal 2 (CD28/B7) progress T cell to the early G(1) phase inducing production of interleukin-2 (IL-2) and other T cell growth factors (TGFs). Signal 3 (cytokine/cytokine receptor) initiate cell division and differentiation in the late G(1)/S phase. Whereas CsA binding to calcineurin blocks Signal 1/2, SRL binding to mammalian target of rapamycin (mTOR) blocks Signal 3. Our preclinical studies have established the in vivo principles of the effects exhibited by SRL alone on allograft survival, synergism between SRL and CsA as well as two drugs pharmacokinetic and pharmacodynamic interactions. In our experimental model, a 14 day i.v. continuous infusion of SRL by osmotic pump into rat recipients extended the survivals of heart allografts in a dose-dependent fashion. In comparison to untreated controls (MST of 6.3 +/- 0.5 days), 0.08 mg/kg SRL extended MST to 34.4 +/- 12.1 days, and 0.8 mg/kg to 74.1 +/- 20.2 days, with 6/18 allografts surviving for more than 100 days. Since almost identical results were produced by 10-fold higher SRL doses delivered by oral gavage, we estimated its bioavailability at 10%. Similarly, SRL prolonged the survivals of kidney, pancreas, and small bowel allografts in rats. At the same time large animal models cautioned about potential toxicities, namely intestinal vasculitis. The synergistic interactions of CsA and SRL may be explained by sequential effects in the early G(0)/G(1) versus late G1/S phases of cell cycle progression, respectively. The in vivo interaction of SRL with other immunosuppressive drugs was evaluated by the median effect analysis and the combination index (CI) values (CI = 1 shows additive, CI < 1, synergistic, and CI > 1, antagonistic, interactions). Oral SRL proved to be synergistic in both CsA-resistant mouse (CI = 0.4-1.5) and CsA-sensitive rat (CI = 0.3-0.6) models. The pharmacokinetic interactions of SRL and/or CsA were evaluated in rats for i.v. and oral formulations. Although low CsA and SRL i.v. doses did not affect each other levels, potent interaction was observed after oral gavage: CsA increased SRL levels by 2-11 folds; and, SRL increased CsA levels by 2-3-folds. Our results suggested that both pharmacodynamic and pharmacokinetic interactions contribute to the synergism between SRL and CsA. We also estimated the impact of CsA/SRL interaction on renal dysfunction, myelosuppression, and hyperlipidemia. Salt depleted rats treated with SRL (0.4-6.4 mg/kg) and/or CsA (2.5-20 mg/kg) were examined for glomerular filtration rates (GFR), lipid levels, and bone marrow cellularity. CsA-induced kidney function deficiency was exacerbated by SRL. This exacerbation of renal dysfunction correlated with increased CsA levels in kidneys when combined with SRL. Furthermore, CsA potentiated SRL-mediated toxicities, namely myelosuppression and increased cholesterol. In conclusion, SRL therapy is synergistic with CsA but both drug levels should be carefully monitored to avoid toxic effects. PMID- 12742500 TI - Molecular actions of sirolimus: sirolimus and mTor. AB - Recent therapeutic strategies to combat organ allograft rejection have focused on T-cell signaling pathways and the molecules that comprise them. The macrolide antibiotic produced by the bacterium Streptomyces hygroscopicus, known as sirolimus or rapamycin, has shown great therapeutic potential in the transplant setting. Sirolimus alone or in combination with other immunosuppressive agents can block acute rejection, chronic graft destruction, and promote permanent allograft acceptance. Sirolimus targets a unique serine-threonine kinase, mammalian target of rapamycin (mTor), which plays a key role in mitogenic and nutritional cells signals. Within T cells, mTor regulates a number of proteins likely dependent on T cell growth factors such as interleukin 2. This review is focused on the molecular mechanisms by which mTor may regulate T-cell signaling cascades and affect T-cell responsiveness, and how sirolimus likely uncouples this activity. PMID- 12742501 TI - Rapamycin: signaling in vascular smooth muscle. AB - Rapamycin (sirolimus) was initially developed as an antibiotic, then as an immunosuppressant, and recently has been identified as one of the most promising novel agents for prevention of coronary artery stent restenosis. The story of how rapamycin was developed for the prevention of stent restenosis involves the discovery of its antiproliferative and antimigratory actions in vascular smooth muscle and ultimately the demonstration that it inhibits neointimal hyperplasia in a large animal model of restenosis. Rapamycin upregulates the cyclin-dependent kinase inhibitor p27(kip1), resulting in cell-cycle arrest at the G1 to S transition. Rapamycin also inhibits other important cellular functions, including protein translation. The precise mechanisms underlying rapamycin's actions have not been fully elucidated. However, its ability to potently inhibit vascular smooth muscle cell migration and proliferation has been the basis for developing rapamycin-eluting coronary artery stents that have reduced in-stent restenosis from about 30% to less than 5% in large clinical trials. PMID- 12742502 TI - Tail profile: a more accurate system for analyzing DNA damage using the Comet assay. AB - The Comet assay (single cell gel electrophoresis assay) measures DNA strand breaks in individual cells. In the assay cells are embedded in agarose, lysed, and electrophoresed under low voltage, allowing migration of damaged DNA. The DNA is stained and subsequently viewed with an epifluorescent microscope. If DNA damage has occurred the electrophoresed DNA fragments appear as a diffuse tail behind the nucleus known as a "comet". Many computer-aided analysis systems are currently in use to quantify the amount of DNA damage that is represented by a comet image. Here, we present a novel method of analysis known as "tail profile". This method of analysis provides several advantages over currently employed methods, which rely primarily on the "tail moment" method of analysis. We compared the amount of DNA damage reported from both the tail profile and tail moment methods of analysis and observed a 26% (P<0.0001) increase in damage detected by tail profile across the 10-25 microm range of tail length, where the majority of the relevant comet data is concentrated. We further report that this increase in sensitivity is not only limited to assessing DNA damage, but also to gathering data from DNA repair assays. Furthermore, we demonstrate increased functionality and extended data analysis capabilities with the use of a compressed collection of images called a "comet chip" and through a visual representation of data called a "profile plot". Use of the custom macros enabled us to detect an unexpected characteristic of the electrophoretic profile, giving us novel insight into the nature of comet analysis. In addition to the increased analytical sensitivity proffered by this system, the tail profile macros are upgradeable and platform independent. PMID- 12742503 TI - Evaluation of hyperdiploidy in the bladder epithelial cells of male F344 rats treated with ortho-phenylphenol. AB - Ortho-phenylphenol (OPP) is a broad-spectrum fungicide and anti-bacterial agent that has been shown to cause bladder cancer in male F344 rats. An earlier study to investigate the potential role of aneuploidy in OPP-induced bladder carcinogenicity, failed to detect increases in frequencies of hyperdiploidy/polyploidy in treated animals, presumably due to the presence of polyploid cells in the bladder. To overcome this problem, we utilized a novel approach to determine increases in numerical alterations in the slowly dividing replicating cells of the rat bladder following treatment with OPP. Collagenase digestion of the bladder was used to enrich for actively-dividing cells and FISH in conjunction with BrdU was employed to detect hyperdiploidy in the replicating interphase cells. Initial studies were performed using FISH with a chromosome 4 probe. Follow-up studies were conducted with OPP and a positive control, vinblastine sulfate using probes for chromosomes 4 and 19. No significant increases in hyperdiploidy/polyploidy were seen in the replicating bladder cells of the OPP-treated rats using FISH with either the chromosome 4 or 19 probes. As expected, no significant increases in hyperdiploidy were seen in the non replicating cells. In contrast, highly significant increases in hyperdiploidy/polyploidy, as detected using FISH with probes for either chromosome 4 or 19, were seen in the replicating cells from rats treated with a combination of OPP and vinblastine. The inability to detect increases in hyperdiploidy/polyploidy in the bladder of OPP-treated rats indicates that chromosome gain is unlikely to play a major role in the early genotoxic effects of OPP. However, the increase in hyperdiploidy/polyploidy induced by vinblastine sulfate in OPP-treated rats, clearly demonstrates that this approach using FISH in combination with BrdU is capable of detecting changes in chromosome number even in slowly-dividing tissues, such as the urinary bladder. PMID- 12742504 TI - Bromate induces loss of heterozygosity in the thymidine kinase gene of L5178Y/Tk(+/-)-3.7.2C mouse lymphoma cells. AB - Potassium bromate (KBrO(3)) induces DNA damage and tumors in mice and rats, but is a relatively weak mutagen in microbial assays and the in vitro mammalian Hprt assay. Concern that there may be a human health risk associated with bromate, a disinfectant by-product of ozonation, has accompanied the increasing use of ozonation as an alternative to chlorination for treatment of drinking water. In this study, we have evaluated the mutagenicity of KBrO(3) and sodium bromate (NaBrO(3)) in the Tk gene of mouse lymphoma cells. In contrast to the weak mutagenic activity seen in the previous studies, bromate induced a mutant frequency of over 100 x 10(-6) at 0.6mM with minimal cytotoxicity (70-80% survival) and over 1300 x 10(-6) at 3mM ( approximately 10% survival). The increase in the Tk mutant frequency was primarily due to the induction of small colony of Tk mutants. Loss of heterozygosity (LOH) analysis of 384 mutants from control and 2.7 mM KBrO(3)-treated cells showed that almost all (99%) bromate induced mutants resulted from LOH, whereas in the control cultures 77% of the Tk mutants were LOH. Our results suggest that bromate is a potent mutagen in the Tk gene of mouse lymphoma cells, and the mechanism of action primarily involves LOH. The ability of the mouse lymphoma assay to detect a wider array of mutational events than the microbial or V79 Hprt assays may account for the potent mutagenic response. PMID- 12742505 TI - Evaluation of cytological effects of Zn2+ in relation to germination and root growth of Nigella sativa L. and Triticum aestivum L. AB - The effects of different treatments with zinc sulfate (Zn(2+)) on the cytology and growth of Nigella sativa and Triticum aestivum were investigated. Five concentrations of zinc sulfate ranging from 5 to 25mg/l were applied for 6, 12, 18, and 24h. The treatments reduced the germination percentages of N. sativa seeds and T. aestivum grains and inhibited the root growth of both plants. Concentrations higher than 25mg/l of Zn(2+) applied for 24h were toxic for both plants. The non-lethal concentrations of Zn(2+) showed an inhibitory effect on cell division in root tips of both plants and caused a decrease in their mitotic index values. The reduction in MI in root tips of T. aestivum was more evident than that of N. sativa. All treatments changed the frequency of mitotic phases as compared with the control values. The total percentage of abnormalities in N. saliva was more than that in T. aestivum. Zn(2+) treatments produced a number of mitotic abnormalities in dividing cells in root tips of both plants resulting from its action on the spindle apparatus such as C-metaphases, lagging chromosomes and multipolar anaphases and telophases. Also, Zn(2+) induced vacuolated nuclei and irregular prophases. The induction of chromosomal stickiness and chromosomal aberrations such as bridges and breaks indicates its action on the chromosome. These abnormalities (chromosome breaks and chromosomal bridges at ana-telophases) indicate true clastogenic potential of the ions tested. PMID- 12742506 TI - Genotoxicity of two anticancer drugs, gemcitabine and topotecan, in mouse bone marrow in vivo. AB - In this study, the genotoxic effects of gemcitabine and topotecan were investigated in mouse bone marrow cells using the micronucleus and chromosomal aberration test systems. Gemcitabine increased the frequency of micronuclei, particularly at the median dose for the 24-, 36-, and 48-h sampling intervals. It had cytotoxic effects on the bone marrow and decreased the polychromatic/normochromatic erythrocyte ratio dose-dependently for all sampling intervals. Gemcitabine significantly decreased the mitotic index at the 24-h time point. It increased the number of abnormal cells and induced a significant increase in total chromosomal aberrations. For the 6-h sampling time, gemcitabine neither induced chromosomal aberrations nor reduced the mitotic index. Topotecan also induced high levels of micronuclei, particularly for the 24- and 36-h sampling times and it decreased the polychromatic/normochromatic erythrocyte ratio for all sampling intervals, which is indicative of bone marrow cytotoxicity. The bone marrow metaphase analysis showed that topotecan significantly elevated the number of abnormal metaphases and total chromosomal aberrations at 6 and 24h, in a dose-dependent manner. It also decreased the mitotic index for both sampling intervals. In conclusion, the results of this study indicate that the two chemotherapeutics gemcitabine and topotecan have cytotoxic and genotoxic effects in mouse bone marrow. PMID- 12742507 TI - Bulky DNA adducts in human sperm: relationship with fertility, semen quality, smoking, and environmental factors. AB - The integrity of DNA of spermatogenic cells can be affected by endogenous and exogenous genotoxic factors. Resulting DNA damage in spermatozoa may significantly contribute to impaired fertility. Here, the 32P-postlabeling method was used to analyze the levels of bulky DNA adducts in sperm cells in a group of 179 males, either healthy donors or patients with an impaired fertility. When all donors were analyzed, the levels of bulky DNA adducts was 1.2-fold higher in smokers than in non-smokers, but the difference was not statistically significant (P=0.054). However, a statistically significant difference existed between current smokers and never smokers among the healthy individuals (1.7-fold increase, P=0.008). No correlation between alcohol or coffee consumption and sperm DNA adducts was found. The levels of DNA adducts in sperm seemed to be unaffected by environmental and occupational factors. On the other hand, groups of healthy persons and patients with male-factor infertility differed significantly with respect to the level of bulky DNA adducts (P=0.012). A significant negative correlation between DNA adducts and sperm concentration or sperm motility existed among patients with an impaired fertility (n=93; P<0.029, r(S)=-0.225). These results suggest that DNA adducts in sperm cells can be applied as potential biomarkers in studies of human infertility. PMID- 12742509 TI - Effects of temperature on baseline and genotoxicant-induced DNA damage in haemocytes of Dreissena polymorpha. AB - The potential application of the Comet assay for monitoring genotoxicity in the freshwater mussel Dreissena polymorpha was explored and a preliminary investigation was undertaken of the baseline levels of DNA damage in mussel haemocytes of animals kept at different temperatures. In addition, in vitro cell sensitivity against genotoxicants was assessed in relation to increasing temperatures. The mussels were kept at four different constant temperatures (4, 18, 28 and 37 degrees C) for 15 h. The haemocytes withdrawn were treated in vitro with melphalan, as a model genotoxic compound, or sodium hypochlorite, a common water disinfectant capable of producing mutagenic/carcinogenic by-products, at the established temperatures for 1h. The data obtained in vivo, in cells directly withdrawn from the mussels showed a significant (P<0.001, Student's t test) inter individual variability, probably due to genetic and epigenetic factors and an increasing amount of DNA damage at increasing temperature. Mussel haemocytes showed a clear dose-response effect after in vitro melphalan treatment. Hypochlorite treatment also significantly increased DNA migration: the damage was temperature dependent, with a similar increase at 4 and 28 degrees C and a minimum level at 18 degrees C. This study demonstrates the potential application of the Comet assay to haemocytes of D. polymorpha. However, these findings suggest that temperature could alter both DNA damage baseline levels in untreated animals and cell sensitivity towards environmental pollutants in in vitro conditions. Therefore, more information is needed about seasonal variations and the natural background levels of DNA damage in mussels living in the wild, before they are used for the monitoring of genotoxic effects in aquatic environments. PMID- 12742508 TI - D-Methylphenidate is non-genotoxic in in vitro and in vivo assays. AB - D-Methylphenidate (dexmethylphenidate; D-MPH) and its racemate D,L methylphenidate (D,L-MPH) are currently prescribed for the chronic treatment of attention deficit hyperactivity disorder (ADHD) in children. Studies have shown that D-MPH is the pharmacologically active enantiomer for ADHD and is therefore the preferred drug for the treatment of ADHD symptoms. Although studies on the mutagenicity of D,L-MPH have been conducted, similar data for D-MPH are lacking. Therefore, D-MPH was evaluated in the bacterial reverse mutation and mouse lymphoma assays with and without S9 and in a bone marrow micronucleus test in male and female CD-1 mice. As a comparison, the L-enantiomer and racemate were also included in the assessments. While MPH-associated toxicity was observed in the mammalian tests, none of the three compounds tested induced mutagenic or clastogenic effects. Our present results along with published epidemiological data from patient populations are consistent with the conclusion that D-MPH and D,L-MPH do not present a carcinogenic risk to humans. PMID- 12742510 TI - Exposure to acrylonitrile induced DNA strand breakage and sex chromosome aneuploidy in human spermatozoa. AB - To explore acrylonitrile (ACN)-induced DNA strand breakage and sex chromosome aneuploidy in human spermatozoa, semen parameters were examined among 30 acrylonitrile-exposed workers according to WHO laboratory manual for the examination of human sperm. DNA strand breakage of sperm cells was investigated among 30 ACN-exposed workers using single cell gel electrophoresis (SCGE). The frequency of sex chromosome aneuploidy in sperm cells was analyzed among nine ACN exposed workers using fluorescence in situ hybridization (FISH). The geometrical mean of sperm density was 75 x 10(6)ml(-1) in exposure group, significantly lower than 140 x 10(6)ml(-1) in the control. The geometrical mean of sperm number per ejaculum was 205 x 10(6) in exposure group, significantly lower than 280 x 10(6) in the control. The rates of comet sperm nuclei were 28.7% in exposure group, significantly higher than 15.0% in the control. Mean tail length was 9.8 microm in exposure group, longer than 4.3 microm in the control. The frequency of sex chromosome disomy was 0.69% in exposure group, significantly higher than 0.35% in the control. XY-bearing sperm was the most common sex chromosome disomy, with an average rate of 0.37% in exposure group, and 0.20% in the control. XX- and YY bearing sperm accounted for an additional 0.09 and 0.23% in exposure group, and 0.05 and 0.10% in the control. The results indicate that ACN affect semen quality among ACN-exposed workers. ACN or its metabolites could induce reproductive defects as an in vivo multipotent genotoxic agent by inducing DNA strand breakage and sex chromosome non-disjunction in spermatogenesis. PMID- 12742511 TI - Correlation of antimutagenic activity and suppression of CYP1A with the lipophilicity of alkyl gallates and other phenolic compounds. AB - Alkyl gallates are widely used as food antioxidants. Methyl, ethyl, propyl, lauryl, and cetyl gallates showed antimutagenicity to activated 2-aminoanthracene (2AA)-induced SOS responses in Salmonella typhimurium TA1535/pSK1002. They also exhibited a suppressive effect on 3-methylcholanthrene (3-MC)-induced cytochrome P450 1A (CYP1A) in human hepatoma HepG2 cells, as indexed by the 7 ethoxyresorufin-O-deethylase (EROD) activity, and on CYP1A protein level. Both antimutagenicity and suppression of CYP1A appeared to be dependent on alkyl chain lengths, which suggested lipophilicity dependence. Based on those results, we investigated 26 other phenolic compounds for their lipophilicity, antimutagenicity and inhibition of EROD activity. The lipophilicity correlated well with the inhibition of EROD activity (r=0.78), and the inhibition of EROD activity correlated with the antimutagenicity of those compounds (r=0.71). The results suggest that the lipophilicity of the phenolic compounds may be an important factor in their ability to inhibit EROD activity. PMID- 12742512 TI - Genotoxicity of hydrated sulfur dioxide on root tips of Allium sativum and Vicia faba. AB - Genotoxicity of sulfur dioxide (SO(2)) and its hydrates (bisulfite and sulfite) in human lymphocytes and other mammalian cells have been found earlier in our laboratory. In the present studies, we used Allium stavium and Vicia faba cytogenetic tests, which are the highly sensitive and simple plant bioassays. A mixture of sodium bisulfite and sodium sulfite (1:3), at various concentrations from 1 x 10(-4) to 2 x 10(-3)M was used for the treatment. Genotoxicity was expressed in terms of anaphase aberration (AA) frequencies in the Vicia-AA test and in terms of micronuclei (MCN) frequencies in both Vicia-MCN test and Alllium MCN test. On average, the results showed a 1.7-3.9-fold increase of AA frequencies and a 3.5-4.5-fold increase of MCN frequencies in Vicia root tips as compared with the negative control. Similarly, results of Allium-MCN test also showed a significant increase in MCN frequencies in the treated samples. In addition, pycnotic cells (PNC) appeared in Allium root tips of treated groups. The frequencies of MCN, AA and PNC increased dose-dependently and the cell cycle delayed at the same time in bisulfite treated samples. Results of the present study suggest that the Vicia and Allium cytogenetic bioassays are efficient, simple and reproducible in genotoxicity studies of bisulfite. PMID- 12742513 TI - Relative percent cell survival and positive response in the in vitro micronucleus test. PMID- 12742514 TI - Effects of propofol on GABAA channel conductance in rat-cultured hippocampal neurons. AB - Channels were activated, in ripped-off patches from rat-cultured hippocampal neurons, by propofol alone, propofol plus 0.5 microM GABA (gamma-aminobutyric acid) or GABA alone. The propofol-activated currents were chloride-selective, showed outward-rectification and were enhanced by 1 microM diazepam. The maximum propofol-activated channel conductance increased with propofol concentration from less than 15 pS (10 microM) to about 60 pS (500 microM) but decreased to 40 pS in 1 mM propofol. Fitting the data from 10 to 500 microM propofol with a Hill-type equation gave a maximum conductance of 64 pS, an EC50 value of 32 microM and a Hill coefficient of 1.1. Addition of 0.5 microM GABA shifted the propofol EC50 value to 10 microM and increased the maximum channel conductance to about 100 pS. The Hill coefficient was 0.8. The maximum channel conductance did not increase further when 1 microM diazepam was added together with a saturating propofol concentration and GABA. The results are compared to effects other drugs have on GABAA channels conductance. PMID- 12742515 TI - Effects of ginsenoside on G protein-coupled inwardly rectifying K+ channel activity expressed in Xenopus oocytes. AB - Recently, we provided evidence that ginsenoside, the active component of Panax ginseng, uses the pertussis toxin-insensitive Galpha(q/11)-phospholipase C-beta3 signal transduction pathway to increase Ca(2+)-activated Cl(-) currents in the Xenopus oocyte. Other investigators have shown that stimulation of receptors linked to the Galpha(q)-phospholipase C pathway inhibits the activity of G protein-coupled inwardly rectifying K(+) (GIRK) channels. In the present study, we sought to determine whether ginsenoside influenced the activity of GIRK 1 and GIRK 4 (GIRK 1/4) channels expressed in the Xenopus oocyte, and if so, the underlying signal transduction mechanism. In oocytes injected with GIRK 1/4 channel cRNA, bath-applied ginsenoside inhibited the high K(+) solution-elicited GIRK current (EC(50): 4.9+/-4.3 microg/ml). Pretreatment of the oocyte with pertussis toxin reduced the high K(+) solution-elicited GIRK current by 49%, but it did not alter the inhibitory effect of ginsenoside on the GIRK current. Prior intraoocyte injection of cRNA(s) coding Galpha(q), Galpha(11) or Galpha(q)/Galpha(11), but not Galpha(i2) or Galpha(oA), attenuated the inhibitory ginsenoside effect. Injection of cRNAs coding Gbeta(1)gamma(2) also attenuated the ginsenoside effect. Preincubation of GIRK channel-expressing oocytes with phospholipase C inhibitor, [1-[6-((17b-3-Methoxyestra-1,3,5(10)-trien-17 yl)amino)hexyl]-1H-pyrrole-2,5-dione] (U73122), or protein kinase C inhibitor, staurosporine or chelerythrine, blocked the inhibitory ginsenoside effect on the GIRK current. Intraoocyte injection of bis (o-aminophenoxy)ethane-N,N,N',N' tetracetic acid (BAPTA), a free Ca(2+) chelator, had no significant effect on the action of ginsenoside. Taken together, these results suggest that ginsenoside inhibits the activity of the GIRK 1/4 channel expressed in the Xenopus oocyte through a pertussis toxin-insensitive and Galpha(q/11)-, phospholipase C- and protein kinase C-mediated signal transduction pathway. PMID- 12742516 TI - S-15176 inhibits mitochondrial permeability transition via a mechanism independent of its antioxidant properties. AB - Mitochondrial Ca(2+) accumulation can induce a sudden increase in the permeability of the inner membrane. This phenomenon is due to the generation of a large nonselective ion channel, termed the permeability transition pore (PTP), which contributes to cellular injury during ischemia and reperfusion. Inhibition of PTP generation constitutes a relevant pharmacological target to protect a cell from death. In this study, we examined the effect of S-15176 ((N-[(3,5-di tertiobutyl-4-hydroxy-1-thiophenyl)]-3-propyl-N'-(2,3,4 trimethoxybenzyl)piperazine), a novel anti-ischemic agent, on PTP in rat liver mitochondria. S-15176 prevented PTP opening generated by various triggering agents, as attested by the concentration-dependent inhibition of mitochondrial swelling, of mitochondrial membrane potential dissipation and of NADPH oxidation. These effects were associated with an increase in the Ca(2+) loading capacity of mitochondria. S-15176 was a strong inhibitor of lipid peroxidation, but experiments with another trimetazidine derivative devoid of antioxidant activity indicated that this activity was not essential to the inhibitory effect. Binding studies demonstrated that [3H]S-15176 bound to mitochondrial binding sites, especially those localized in the inner membrane. These sites were shared by several well-known inhibitors of PTP opening. These results demonstrate that the mechanism by which S-15176 protects mitochondria against the deleterious effects of ischemia-reperfusion involves inhibition of PTP opening and provide evidence that the drug operates through low structural specificity binding sites located in the inner mitochondrial membrane. PMID- 12742517 TI - Potentiation of metabotropic GABAB receptors by L-amino acids and dipeptides in rat neocortex. AB - Selected neutral L-alpha-amino acids, and their dipeptides, were reversible, stereospecific, potentiators of GABA(B) receptor-mediated hyperpolarizing responses to baclofen (3-100 microM) in rat neocortical slices. These responses were sensitive to the GABA(B) receptor antagonist (+)-(S)-5,5-dimethylmorpholinyl 2-acetic acid (Sch50911) (30 microM). Most potent were L-Leu, L-Ile and L-Phe, as were the dipeptides L-Phe-Phe and L-Phe-Leu, and less potent were L-Met, L-Val, L Cys, L-Cystine, L-Tyr, L-Thr, L-Arg and L-Ser. Inactive were L-Trp, L-His, L-Lys and L-Pro. These potentiators gave leftward shifts of the baclofen concentration response curves with a Hill slope of 2, and a marked increase in the maximal hyperpolarizing responses. Selected L-amino acids and dipeptides are a class of naturally occurring GABA(B) potentiators, which may be allosteric modulators. PMID- 12742519 TI - Risperidone reduces limited access alcohol drinking in alcohol-preferring rats. AB - An atypical antipsychotic drug risperidone reduced ethanol drinking of ethanol preferring Alko, Alcohol (AA) rats in a limited access paradigm. Its effect was transient at a dose known to preferentially antagonize the 5-HT(2) receptors (0.1 mg/kg, s.c.), but long-lasting when the dose was increased to 1.0 mg/kg that also blocks dopamine D(2) receptors. Risperidone also reduced dose-dependently locomotor activity and limited access saccharin intake of the AA rats, indicating that its effect on ethanol drinking was not selective. Risperidone at 0.1 mg/kg given before four successive daily ethanol-drinking sessions significantly reduced the ethanol intake. These data from an animal model of high ethanol intake suggest that risperidone should be tested in various populations of alcoholics for reducing ethanol consumption. PMID- 12742518 TI - Behavioral effects of rimcazole analogues alone and in combination with cocaine. AB - Several sigma receptor ligands have been reported to also have affinity for the dopamine transporter, among them rimcazole (9-[3-(cis-3,5-dimethyl-1 piperazinyl)propyl]carbazole dihydrochloride). However, rimcazole lacks behavioral effects like those of other dopamine uptake inhibitors, such as cocaine and GBR 12909 (1-(2-[bis(4-fluorophenyl)methoxy]ethyl)-4-(3 phenylpropyl)piperazine dihydrochloride). Because of this profile, the interactions with cocaine of rimcazole and several of its novel analogues were assessed. The compounds studied were rimcazole, its N-methyl analogue, SH 1-73 (9 [3-(cis-3,5-dimethyl-4-methyl-1-piperazinyl)-propyl]carbazole hydrobromide), the dibrominated analogue, SH 1-76 (3,6-dibromo-9-[3-(cis-3,5-dimethyl-1-piperazinyl) propyl]carbazole hydrochloride), and the N-propylphenyl analogues, SH 3-24 ([3 (cis-3,5-dimethyl-4-[3-phenylpropyl]-1-piperazinyl)-propyl]diphenylamine hydrochloride) and SH 3-28 (9-[3-(cis-3,5-dimethyl-4-[3-phenylpropyl]-1 piperazinyl)-propyl]carbazole hydrobromide). The former has a diphenyl-amine group in place of the carbazole moiety of rimcazole, giving the compound additional structural similarity to GBR 12909. The rimcazole analogues produced dose-related decreases in locomotor activity, and also decreased cocaine stimulated activity in mice. In rats trained to discriminate 10 mg/kg cocaine (i.p.) from saline injections, cocaine and GBR 12909 each produced a dose-related increase in cocaine-appropriate responding. Cocaine also increased rates of responding. SH 3-28 decreased cocaine-appropriate responding at the cocaine training dose to about 58% (SH 3-28) with two of five subjects selecting the cocaine response key. Neither rimcazole nor SH 3-24 produced a significant attenuation of the discriminative effects of cocaine. Rimcazole and its analogs all attenuated the increases in rates of responding produced by cocaine. In contrast to effects obtained with rimcazole analogs, GBR 12909 potentiated the cocaine-induced increases in locomotor activity and operant behavior, as well as the discriminative-stimulus effects of cocaine. The present results indicate that analogues of rimcazole can attenuate the behavioral effects of cocaine, and though the mechanism for these effects is not presently clear, it is possible that this attenuation maybe mediated by actions of the rimcazole analogues at the dopamine transporter and/or sigma receptors. PMID- 12742520 TI - Intravenously injected CDP-choline increases blood pressure and reverses hypotension in haemorrhagic shock: effect is mediated by central cholinergic activation. AB - Intravenous (i.v.) administration of cytidine-5'-diphosphate choline (CDP choline) (100, 250 and 500 mg/kg) increased blood pressure in normal rats and reversed hypotension in haemorrhagic shock. Choline (54 mg/kg; i.v.), at the dose equimolar to 250 mg/kg CDP-choline decreased blood pressure of rats in both conditions and caused the death of all hypotensive animals within 2-5 min. Equimolar dose of cytidine (124 mg/kg; i.v.) did not change cardiovascular parameters. Choline levels in plasma, lateral cerebral ventricle and hypothalamus increased after CDP-choline administration. Intracerebroventricular (i.c.v.) hemicholinium-3 pretreatment (20 microg), greatly attenuated the pressor effect of CDP-choline in both conditions. Atropine pretreatment (10 microg; i.c.v.) did not change the pressor effect of CDP-choline while mecamylamine (50 microg; i.c.v.) abolished the pressor response to drug. Besides, acetylcholine (1 micromol; i.c.v.) produced similar increases in blood pressure in normal and hypotensive conditions to that observed in CDP-choline given rats. CDP-choline (250 mg/kg; i.v.) increased plasma catecholamines and vasopressin levels but not plasma renin activity. Pretreatment of rats with either prazosin (0.5 mg/kg; i.v.) or vasopressin V(1) receptor antagonist, [beta-mercapto,beta,beta cyclopentamethylenepropionyl(1),O-Me-Tyr(2)-Arg(8)]vasopressin (10 microg/kg; i.v.), attenuated the pressor response to CDP-choline while simultaneous administration of these antagonists before CDP-choline injection completely blocked the pressor effect. Results show that i.v. CDP-choline increases blood pressure and reverses hypotension in haemorrhagic shock. Activation of central nicotinic cholinergic mechanisms by the increases in plasma and brain choline concentrations appears to be involved in the pressor effect of this drug. Moreover, the increases in plasma catecholamines and vasopressin levels mediate these effects. PMID- 12742521 TI - Inhibitory effects of bromocriptine on corticosterone secretion in male rats. AB - Bromocriptine, a dopamine D2 receptor agonist, is widely used for treating prolactinoma, Parkinson's disease and galactorrhea. However, the influence of bromocriptine on the endocrine system, especially adrenal function, is not clear. The present study was aimed to investigate the effects of bromocriptine on corticosterone production in rats. Male rats were treated or not treated by bromocriptine (5 mg/kg, s.c.) twice per day for 2 days before decapitation. The adrenal zona fasciculata-reticularis cells were prepared and incubated with adrenocorticotropic hormone (ACTH), forskolin (an adenylyl cyclase activator), 8 bromo-adenosine 3':5' cyclic monophosphate (8-Br-cAMP, a membrane-permeable analogue of cAMP), and steroidogenic precursors including 25-OH-cholesterol and pregnenolone. The concentrations of prolactin, corticosterone and pregnenolone in the plasma and/or medium were measured by radioimmunoassay (RIA). The protein expression of cytochrome P450 side-chain cleavage (P450scc) enzyme and steroidogenic acute regulatory protein (StAR) was analyzed by Western blotting. Administration of bromocriptine in vivo resulted in a decrease in the levels of plasma prolactin and corticosterone. Basal--and ACTH--as well as forskolin stimulated corticosterone secretion by zona fasciculata-reticularis cells was also lower in bromocriptine-treated rats than in control animals. The decreased production of corticosterone in zona fasciculata-reticularis cells could be reversed by administration of 8-Br-cAMP. The corticosterone and pregnenolone release induced by 25-OH-cholesterol in zona fasciculata-reticularis cells was reduced by administration of bromocriptine. The protein expression of both StAR protein and P450scc in zona fasciculata-reticularis cells was inhibited in the bromocriptine-treated group. Administration of bromocriptine in vitro reduced the release of corticosterone stimulated by ACTH and forskolin in rat zona fasciculata-reticularis cells. These results suggested that bromocriptine caused adrenal dysfunction through inhibition of ACTH action and of the activity of adenylyl cyclase, and impaired the early steps of corticosterone biosynthesis. PMID- 12742523 TI - Proteomics in gerontological research. AB - The instrumental tools of proteomics offer great potential for molecular studies of biological aging. This article reviews specific applications and emerging concepts of proteomics, which are or can be applied to aging research. PMID- 12742522 TI - Hypoglycemic agent YM440 suppresses hepatic glucose output via gluconeogenesis by reducing glucose-6-phosphatase activity in obese Zucker rats. AB - Using a glucose clamp, we had shown that YM440, (Z)-1,4-bis[4-[(3,5-dioxo-1,2,4 oxadiazolidin-2-yl)methyl]phenoxy]but-2-ene, reduced the increased hepatic glucose output in obese Zucker rats. We further examined effects of YM440 on 14C incorporation from [14C]bicarbonate into blood glucose via gluconeogenesis, and on gluconeogenic enzymatic activities. Fed obese Zucker rats showed a 4-fold increase of 14C-incorporation into blood glucose compared to that in lean rats. Glucose-6-phosphatase and fructose-1,6-bisphosphatase activities in obese rats were increased 1.4-fold and 1.6-fold compared with lean rats. YM440 (300 mg/kg for 2 weeks) decreased 14C-incorporation into blood glucose by 29% in obese rats. Glucose-6-phosphatase but not fructose-1,6-bisphosphatase activity was reduced by YM440 and closely correlated with 14C-incorporation into blood glucose, indicating a key role for glucose-6-phosphatase in hepatic glucose output. These results suggest that the increased gluconeogenesis in obese rats is mainly due to the increased activities of glucose-6-phosphatase and fructose-1,6-bisphosphatase and that YM440 suppresses hepatic glucose output by reducing glucose-6 phosphatase activity. PMID- 12742524 TI - The Kronos longevity research institute. AB - The Kronos Longevity Research Institute (KLRI), founded in Jan., 2000, is a Phoenix-based not-for-profit institution conducting clinical translational research aimed at early detection and prevention of age-related diseases and slowing or reversing the aging process. KLRI also provides education in biomedical gerontology for regional and national professional and lay communities. KLRI is privately funded, mainly by the Aurora Foundation. S. Mitchell Harman, M.D., Ph.D., founding Director and President is board-certified in internal medicine and endocrinology and a former section chief and acting clinical director of the National Institute on Aging, NIH, with an international reputation as a leader in the field of hormones and aging. Other professional staff are: a Clinical Director, Director of Exercise Sciences, Senior Scientist, and Clinical Study Coordinator. KLRI's facility includes a clinical study center (CSC), an exercise study center (ESC), and a molecular laboratory. Current research focuses on relationships among aging, endocrine function, oxidative stress, and sarcopenia. All research projects are pre-reviewed by KLRI's Scientific Advisory Board, a distinguished group of biomedical investigators. KLRI sponsors a series of bimonthly seminars in Phoenix and an annual two-day national symposium, with talks on biomedical gerontology presented by world renowned experts. The institute has plans to double faculty, staff, and research activities by 2006, which will require new sources of funding. The aging demography of the first half of the century will make KLRI's research increasingly relevant to the population of the U.S. and the world. PMID- 12742525 TI - Age-related apoptotic dysfunction: any role for polypeptide aspartate instability? PMID- 12742526 TI - Altered methionine metabolism in long living Ames dwarf mice. AB - Ames dwarf mice (df/df) are deficient in growth hormone, prolactin, and thyroid stimulating hormone and live significantly longer than their normal siblings. In the current study, we found that the hormone deficiencies affect methionine metabolism. We previously reported that the dwarf mice exhibit enzyme activities and levels that combat oxidative stress more efficiently than those of normal mice. Moreover, methionine or metabolites of methionine are involved in antioxidative processes. Thus, we performed an experiment that compared various parameters of methionine metabolism between 18-month old male dwarf (N=6) and wild type (N=5) mice. The specific activity of liver methionine adenosyltransferase (MAT) was significantly elevated (205%, p<0.0001) in the dwarf mice, as were cystathionine synthase (50%, p<0.01), cystathionase (83%, p<0.001), and glycine N-methyltransferase (GNMT, 91%, p<0.001) activities. Even though the activities of MAT and GNMT were elevated, the concentration of liver S adenosylmethionine was decreased (24%, p<0.001) and S-adenosylhomocysteine increased (113%, p<0.001) in the dwarf mice. These data indicate that dwarf mice, compared to wild type mice, have a markedly different metabolism of methionine. Altered methionine metabolism may partially explain earlier reports indicating less oxidative damage to proteins in dwarf mice. Taken together, the data suggest that methionine metabolism may play a role in oxidative defense in the dwarf mouse and should be studied as a potential mechanism of extended lifespan. PMID- 12742527 TI - Age-associated changes in the serum level of apolipoproteins A-I and A-IV and the gene expression as revealed by fasting and refeeding in mice. AB - Alterations of serum apolipoproteins A-I (apo A-I) and A-IV and their mRNAs in young and old mice by fasting and refeeding were investigated by polyacrylamide gel elecrophoresis and Northern blot, respectively. After fasting for three days, serum apo A-I concentration in young mice (6-9 month-old) was increased about 1.5 fold while that of old animals (25-34 month-old) did not change significantly. Apo A-I mRNA was increased about 3-fold and 1.7-fold in the liver and small intestine of the young mice, respectively. The increase in old animals was not more than 1.5-fold in both tissues. The serum apo A-IV was elevated 2-fold and its mRNA was markedly (ca. 50-fold) induced in the liver of fasted young mice, whereas the increase of the mRNA was less than 2-fold in the small intestine. In contrast, induced levels of the protein in serum and its mRNA in both tissues were much less in old mice. In view of the roles of apo A-I and A-IV in triglyceride mobilization and reverse cholesterol transport, the present findings suggest that the reduced induction of the mRNAs for these apolipoproteins in the liver by prolonged fasting and possibly under normal feeding conditions can be an important factor in the impaired immobilization of lipid in old animals, and may, in turn, have implication in age-related diseases such as coronary, cerebral and other vascular disorders. PMID- 12742528 TI - Gender difference in glutathione metabolism during aging in mice. AB - Oxidative damage of the macromolecules increases with age and has been suggested to contribute importantly to the aging process and the pathogenesis of many age related diseases. However, what causes such an increase in the oxidative damage of the macromolecules and whether male and female have the same susceptibility are not clear. In this study, we demonstrated for the first time that although the concentrations of GSH were similar between young male and female mice in most tissues examined and GSH content declined with age in both genders, male mice seemed to experience more dramatic age-associated change in GSH content than did female mice in many tissues. The age-related decline in the GSH content in both male and female mice was also associated with a decrease in the amounts of glutamate cysteine ligase (GCL) mRNAs and proteins as we have reported previously in male rats, further suggesting an important role of GCL in maintaining GSH homeostasis during the aging process. The results from this study may reveal an important basis underlying the gender-associated differences in the longevity and the susceptibility to certain age-related diseases, and also further suggest that the decreased synthesis, which is mainly due to the down regulation of GCL gene expression, may be responsible for the age-associated decline in GSH content. PMID- 12742529 TI - Ageing-related changes in the in vivo function of rat liver macroautophagy and proteolysis. AB - Autophagy is a universal, highly regulated mechanism responsible for the degradation of long-lived proteins, cytomembranes and organelles during fasting and may be the cell repair mechanism that mediates the anti-ageing effects of calorie restriction (Bergamini and Gori, 1995). The function of autophagy was studied in vivo on male Sprague Dawley rats fed ad libitum or 40% food restricted. Autophagy was induced in overnight fasted rats by the injection of an anti-lipolytic agent and was investigated by electron microscopy. Changes in regulatory plasma nutrients and hormones were assessed and rate of proteolysis was calculated from the release of 14C(6)-valine from pre-labelled resident proteins. Results in rats fed ad libitum showed that autophagic-proteolytic response to antilypolitic agents was paramount in one month-old rats; was high but delayed in 2 month-old rats, decreased remarkably in 6 month-old rats and almost negligible at older age. Parallel ageing-related changes were observed in the effects of treatment lowering glucose and insulin plasma levels. Calorie restriction prevented all changes. In view of the known suppressive effects of insulin, it may be concluded that the age-changes of autophagy are secondary to the ageing-related alteration in glucose metabolism and hormone levels, whose appearance is delayed by calorie restriction. Data may support the hypothesis that ad libitum feeding accelerates the rate of ageing by raising insulin plasma levels and suppressing autophagy and membrane maintenance, and that calorie restriction may break this vicious circle. PMID- 12742530 TI - Recovery of slow skeletal muscle after injury in the senescent rat. AB - We studied the contractile, histological and biochemical characteristics of regenerating slow (soleus) muscles of aged rats and the effect of IGF-1 treatment on these parameters. Regenerating soleus muscles were studied 21 days after myotoxic injury. In senescent rats (24 month old), the in situ isometric maximal tetanic force (P0), resistance to fatigue (T20%P0) and shortening speed with an afterload of 20%P0 (SS20%P0) were lower (p<0.05) in regenerating soleus muscles as compared to uninjured controlateral soleus muscles. Moreover, the expression of type 1 myosin heavy chain (MHC-1) was decreased by injury in the soleus muscles of senescent rats (p<0.05). Furthermore, a single injection of IGF-1 (3 microg) into the soleus of senescent rats only slightly increased the level of sarcoplasmic reticulum type 2 Ca(2+)-ATPase in regenerating soleus muscles (p<0.01). Contrary to senescent animals, regenerating soleus of adult rats (10 month old) did not present significantly lower P0 and MHC-1 expression than uninjured controlateral muscles (p>0.05). In conclusion, the regeneration of a slow muscle is more uncompleted 3 weeks after myotoxic injury in senescent rats than in adult rats. It cannot be made more effective by a single injection of IGF 1 into the senescent slow muscle. PMID- 12742531 TI - Modulation of glutathione and thioredoxin systems by calorie restriction during the aging process. AB - Accumulating evidence strongly suggests that oxidative stress underlies aging processes and that calorie restriction (CR) retards aging processes, leading to an extended lifespan for various organisms. Recent studies revealed that the anti aging action of CR depends on its anti-oxidative mechanism. However, at present, the status of glutathione (GSH) and thioredoxin (Trx) system, two major thiol redox systems in animal cells during aging and its modulation by CR has not fully been explored. The purpose of this study is two-fold: one, to determine whether these two systems in rat kidney are altered as a consequence of aging; two, to determine whether these systems can be modulated by anti-oxidative CR. The results of our study showed that GSH and GSH-related enzyme activities decreased with age in ad libitum (AL)-fed rats, while CR rats consistently showed resistance to decreases in these activities. Data from the present data further showed that while Trx and Trx reductase (TrxR) in cytoplasm decrease with age in AL-fed rats, CR prevents these decreases. In contrast, we also found that the nuclear translocation of the redox regulators, Trx and Ref-1, increase with age, which was suppressed in CR rats. Therefore, increases in nuclear Trx and Ref-1 during aging may result in the up-regulation of redox-sensitive transcription factors, such as NF-kappaB or AP-1, via the interaction of Ref-1 and Trx in a redox-dependent manner. Our conclusion is that a redox imbalance occurs during aging and that redox changes are minimized through the anti-oxidative action of CR. PMID- 12742532 TI - Effects of caloric restriction on gene expression along the epididymis of the Brown Norway rat during aging. AB - Aging in the Brown Norway (BN) rat, is associated with changes in the male reproductive tract. In the epididymis, the site of sperm maturation and storage, aging has profound effects on the histology, biochemistry and gene expression of the tissue. Caloric restriction consistently slows aging and maintains health in aging animals. The goal of this study was to assess the effects of caloric restriction on aging-related changes in gene expression in the BN rat epididymis using cDNA microarrays. Caloric restriction attenuated or reversed age-related gene expression changes throughout the epididymis. In the distal regions of the tissue (corpus and cauda epididymidis), greater than 80% of the gene expression decreases were attenuated by caloric restriction. In the most proximal region of the tissue (initial segment), caloric restriction affected the expression of genes associated with lipid and carbohydrate metabolism. In all regions of the epididymal epithelium, caloric restriction had a dramatic effect on the age related decreased expression of genes associated with protein synthesis and mitochondrial function. These results indicate that caloric restriction lessens age-related mitochondrial dysfunction and improves protein synthetic capability throughout the epididymis. PMID- 12742533 TI - Increased frequency of the 2437T allele of the heat shock protein 70-Hom gene in an aged Irish population. AB - The frequency of the functional polymorphism, T2437C transversion (Met-->Thr), in the HSP 70-Hom gene was investigated within a healthy aged Irish population using oligonucleotide probes. The 2437T polymorphic nucleotide was observed to increase in the elderly, although not attaining statistical significance. The TT genotype was observed to be significantly increased within the Irish aged population (p=0.03), while conversely the TC genotype was significantly decreased in the aged subjects (p=0.01). These findings would support the theory that the change from a Met (non-polar and hydrophobic) residue to a Thr (polar and neutral) residue may disrupt the peptide-binding specificity of HSP 70-Hom and have an effect on its functional efficiency. One postulates that the highly significant p value obtained for the TC genotype may infer that the presence of both the T and the C allele (heterozygosity) resulting in the generation of two different HSP 70 Hom protein species may negatively influence longevity. PMID- 12742534 TI - A maternal line study investigating the 4977-bp mitochondrial DNA deletion. AB - The most frequently reported species of mitochondrial DNA (mtDNA) damage associated with ageing is the 4977-bp 'Common Deletion'. However, recent observations have raised several issues within the deletion debate namely: the significance of the 4977-bp deletion (CD) as a universal DNA marker of ageing and mitochondrial dysfunction; and the possibility for maternal transmission of deletions in humans. Previous attempts at answering these questions have been limited because many investigations have been cross-sectional studies of unrelated individuals. With the unique feature of the maternal inheritance of mtDNA, our study overcomes some of these limitations by investigating the CD in human maternal lines, which represent 21 families spanning four generations. Using a highly sensitive PCR methodology, we identified the presence of the CD in leukocytes from all 71 individuals (age range-8 months-99 years) including all infants and children (n=15) which in addition were free of any known mitochondrial diseases. This is important because the few reports of the CD in infants have been linked to mitochondrial disease. These results question the significance of the CD as a universal DNA marker of ageing and subsequent mitochondrial dysfunction and provide support for the possibility for maternal transmission of deletions. PMID- 12742535 TI - Short-term supplementation with lutein affects biomarkers of lutein status similarly in young and elderly subjects. AB - There is evidence that lutein may protect against age-related macular degeneration, cataract, cancers and cardiovascular diseases, but no data have been published on the effect of age on lutein status. The purpose of this work was to determine whether there are major differences in the status of this carotenoid between young and elderly subjects. Initial lutein status and the effect of a 5-week lutein supplementation (9 mg/d) on the most common markers of lutein status were compared in 12 young (26.9+/-0.8yr) and 17 older subjects (67.3+/-1.1yr). Lutein was measured by HPLC in fasting serum, adipose tissue and buccal mucosa cells (BMC) before and after supplementation. Macular pigment optical density (MPOD), which partly depends on retina lutein concentration, was measured by reflectometry before and after supplementation. Initial lutein status was not significantly different between the two groups, irrespective of the lutein status marker. Plasma and BMC lutein concentrations significantly increased in both groups after lutein supplementation, but not MPOD or adipose tissue lutein. Plasma and BMC responses to lutein supplementation (percent variation from initial values) were not significantly different between the two groups. These results suggest that there is no major effect of age on lutein status in healthy subjects. PMID- 12742536 TI - Association of increased neopterin production with decreased humoral immunity in the elderly. AB - Tetanus toxoid (TT) antibodies of 447 adult persons aged 27-69 years were investigated and analyzed in relationship with the time span since the last vaccination against tetanus as well as the serum concentration of neopterin. Neopterin is a pteridine, which is produced by monocytes/macrophages upon stimulation with the type 1 T cell-derived cytokine interferon-gamma. There was an inverse correlation between serum neopterin and TT antibody concentrations (Spearman's rank correlation coefficient: r(s)=-0.259; p<0.0001) which was even stronger when persons with neopterin concentrations and TT antibodies below the third quartile of the study population were excluded (residual group: n=210; r(s)=-0.718; p<0.0001). The study demonstrates that an immunoregulatory shift towards type 1 immunity as indicated by higher neopterin concentrations coincides with lower TT antibody concentrations in the elderly. PMID- 12742537 TI - Increased prevalence of coeliac disease in autoimmune thyroiditis is restricted to aged patients. AB - Patients with autoimmune thyroiditis (AT) have an increased prevalence of coeliac disease (CD), an immune-mediated enteropathy. It is unknown, however, whether prevalence of CD in AT is affected by age. Sera from 514 patients with AT aged <65 yr (46+/-12 yr), 223 patients with AT aged >or=65 yr (71+/-5 yr), 300 controls aged <65 yr (45+/-12 yr), and 300 controls aged >or=65 yr (71+/-6 yr) were tested for IgA anti-tissue transglutaminase (anti-tTG) and antiendomysial antibodies (EmA). If anti-tTG or EmA IgA were positive, jejunal biopsy was performed to confirm CD diagnosis. Prevalence of CD was significantly higher in patients with AT aged >or=65 yr (3.6%, P=0.024) than in patients with AT aged <65 yr (0.6%), controls aged <65 yr and controls aged >or=65 yr (both 0.3%). Prevalence of CD did not significantly differ across patients with AT aged <65 yr, controls aged <65 yr and controls aged >or=65 yr. In conclusion, prevalence of CD is increased in AT but the association is limited to patients aged 65 years or older. Serological screening including anti-tTG-IgA is recommended in these patients. PMID- 12742539 TI - Exercise and postprandial lipid metabolism: an update on potential mechanisms and interactions with high-carbohydrate diets (review). AB - Endurance trained people exhibit low levels of postprandial lipemia. However, this favorable situation is rapidly reversed with de-training and it is likely that the triglyceride (TG) lowering effects of exercise are mainly the result of acute metabolic responses to recent exercise rather than long-term training adaptations. A large body of evidence suggests that postprandial lipemia can be attenuated following an individual exercise session, with the energy expended during exercise being an important determinant of the extent of TG lowering. Increased lipoprotein lipase-mediated TG clearance and reduced hepatic TG secretion are both likely to contribute to the exercise-induced TG reductions. These changes may occur in response to post-exercise substrate deficits in skeletal muscle and/or the liver. In addition, regular exercise can oppose the hypertriglyceridaemia sometimes seen with low-fat, high-carbohydrate diets. Levels of physical activity should therefore be taken into account when considering nutritional strategies for reducing the risk of cardiovascular disease. PMID- 12742540 TI - Breast and lung cancer are associated with a decrease in blood cell amino acid content. AB - The description of different plasma amino acid profiles for specific types of cancer suggests that the metabolic alterations brought about by each type of tumor determine their own, distinctive profile of plasma amino acids. However, the blood cell pool represents an important percentage of the total amount of amino acids and has been reported to undergo significant changes in several physiological situations, thus raising the question of what effect a situation like cancer could have on amino acid blood compartmentation. We determined the levels of individual amino acids in blood, plasma and blood cell compartment of 14 lung cancer patients, 16 breast cancer patients and the corresponding healthy controls (n = 14 and 18, respectively). Cancer, a situation of increased amino acid demand, was accompanied by a decrease in the amino acid availability, of which the blood cell pool would be the main contributor. Thus, the fact that the blood cell pool reflects more intensely than plasma the changes in amino acid availability and undergoes changes according to the demand of amino acids, reinforces the important role of the cell pool in blood amino acid compartmentation and handling. The profiles of blood amino acids characteristic of different types of tumors that have been proposed by some authors could be extended to other compartments-in addition to the plasma-and even be more informative. PMID- 12742541 TI - Modification of analytical procedures for determining vitamin C enzyme (L gulonolactone oxidase) activity in swine liver. AB - Modifications of the analytical method to determine L-gulono-gamma-lactone oxidase (EC 1.1.8) enzyme activity were conducted in pig liver by evaluating the concentration of added substrate (L-gulono-gamma-lactone), glutathione, and various tissue sample-to-buffer ratios in the incubation mixture. Sampling different liver sites (lobes), the effect of different cooling temperatures of the liver immediately after collection, and the effect of tissue storage length on subsequent enzyme activity were evaluated. Our results demonstrated that 10 mM of substrate added to the reaction media maximized L-gulono-gamma-lactone oxidase enzyme activity, whereas increasing levels of glutathione did not greatly affect enzyme activity. High sample-to-buffer ratios resulted in higher L-gulono-gamma lactone oxidase activities but sample analytical variations and background interferences were greater. A 1:4 tissue sample to buffer ratio (weight:weight) resulted in repeatable values, but the importance of maintaining the same ratio of the two components seems to be critical within an experiment. Expressing L gulono-gamma-lactone oxidase enzyme activity on a liver protein rather than on a liver weight basis also resulted in more consistent results. No difference in liver L-gulono-gamma-lactone oxidase enzyme activities or ascorbic acid concentrations occurred between liver lobes. L-gulono-gamma-lactone oxidase enzyme activity from 0 to 90 day of storage was not affected when tissue samples were immediately frozen in liquid nitrogen, or placed on crushed ice. During a 90 day storage the oxidized form of ascorbic acid (dehydroascorbic acid) decreased (P < 0.01), the reduced (ascorbic acid) form increased (P < 0.01), while total ascorbic acid concentration remained constant. PMID- 12742542 TI - Low zinc intake decreases the lymphatic output of retinol in rats infused intraduodenally with beta-carotene. AB - Previously, we have shown that the lymphatic absorption of retinol is significantly decreased in rats fed a low zinc diet. This study was conducted to determine whether the absorption of beta-carotene also is altered in zinc deficient male rats. The absorption of beta-carotene was estimated by determining the amount of retinol appearing in the mesenteric lymph during intraduodenal infusion of beta-carotene. One group of rats was fed the AIN-93G diet but low in zinc (LZ; 3 mg/kg) and the other was fed the same diet adequate in zinc (AZ; 30 mg/kg). The LZ and AZ rats were trained to meal feed equal amounts of the diets twice daily. At 6 weeks, each rat with lymph cannula was infused via an intraduodenal catheter at 3 ml/h for 8 h with a lipid emulsion containing 65.0 nM beta-carotene, 565.1 microM triolein, 27.8 kBq 14C-triolein (14C-OA), 72 mg albumin, and 396 microM Na-taurocholate in 24 ml PBS (pH 6.7). The lymphatic output of retinol over the 8-h period was significantly lower in LZ rats than in AZ rats. The absorption of 14C-OA also was significantly lower in LZ rats. No significant differences were observed between groups in intestinal beta-carotene 15,15'-dioxygenase, retinal reductase, and retinal oxidase activities. The findings demonstrate that low zinc intake or marginal zinc deficiency significantly lowers the absorption of beta-carotene as estimated by lymphatic retinol output. The results also indicate that the decrease in retinol output in LZ rats is not linked to defects in beta-carotene cleavage and subsequent conversion of retinal to retinol in the intestinal mucosa. This study suggests that zinc status is an important factor determining the intestinal absorption of beta-carotene and hence the nutritional status of vitamin A. PMID- 12742543 TI - Comparison of the contents of the main biochemical compounds and the antioxidant activity of some Spanish olive oils as determined by four different radical scavenging tests. AB - The aim of this study was to compare the contents of the main biochemical compounds and the antioxidant capacity of five Spanish olive oils by four different antioxidant tests and to find out the most valuable oil for disease preventing diets. Fatty acids, sterols and individual antioxidant compounds in Arbequina, Hojiblanca, Extra Virgin, Picual and Lampante Spanish olive oils were determined. Antioxidant activities were done as well using different radical scavenging activities: total radical-trapping antioxidative potential by ABAP (TRAP-ABAP), radical scavenging activity by DPPH (RSA-DPPH), antioxidant assay by beta-carotene-linoleate model system (AA-beta-carotene) and total antioxidant status by ABTS (TAA-ABTS). The highest content of all studied antioxidant compounds (353; 329; 4.6 and 2.7 mg/kg for tocopherols, tocotrienols, polyphenols and o-diphenols, respectively) was found in Extra Virgin oil. Also the highest antioxidant capacity was observed in Extra Virgin oil (668 nM/ml; 29.4%; 40.4% and 2.64 mM TE/kg for TRAP-ABAP, RSA-DPPH, AA- beta-carotene and TAA-ABTS, respectively). The correlation between total phenols and antioxidant capacities measured by four methods was very high, but the highest for the beta-carotene (R = 0.9958). In conclusion, the best method for determination of the antioxidant capacity of olive oils is the beta-carotene test. Extra Virgin olive oil has high organoleptic properties and the highest antioxidant activity. The above-mentioned makes this oil a preferable choice for diseases preventing diets. PMID- 12742544 TI - Nervonic acid is transferred from the maternal diet to milk and tissues of suckling rat pups. AB - Three experiments were designed to investigate the metabolism of dietary nervonic acid (24:1n-9, NA) during reproduction in the rat. The first experiment determined the effect of early development on the sphingomyelin (SM) composition of rat heart and liver tissues. Rats were fed a standard chow diet and the SM fatty acid composition of the hearts and livers were analyzed of 18-20 day old fetuses, 14 day old sucklings and adult rats. The 18:0 content of SM decreases with age, while 23:0 and iso 24:0 increase with age. In the second experiment pregnant rats were fed diets supplemented with either canola, corn or peanut oil to determine the effect of diets high in 24:1n-9 and 24:0 on liver and heart SM at birth and after 14 days of suckling. Pups from the dams fed the corn oil diet had elevated 24:2n-6 in SM from heart and liver at birth, but the content of NA was not altered by dietary fat type. In the third experiment oil mixtures were designed to provide elevated levels of 22:1 and 24:1 (canola-N25), 22:0 and 24:0 (peanut-flax) or <0.01% of these fatty acids (olive-flax) and were supplemented to the diets of lactating rats. Canola-N25 oil supplemented to lactating rats resulted in increased 24:1n-9 and 24:1/24:0 with decreased 22:0 and 24:0 in milk SM relative to the other groups. The SM composition of livers of the suckling rats showed significant changes reflecting the changes in milk SM composition after 6 days of milk consumption. These experiments suggest that dietary NA and is not readily transferred across the placental barrier but does readily cross the mammary epithelium and is incorporated into milk SM. In addition, NA in milk appears to cross the intestinal epithelium where it is incorporated into the SM of heart and liver of suckling rats. PMID- 12742546 TI - Induction of hepatic thioredoxin reductase activity by sulforaphane, both in Hepa1c1c7 cells and in male Fisher 344 rats. AB - Sulforaphane (SF), a glucosinolate-derived isothiocyanate found in cruciferous vegetables, is considered an anticarcinogenic component in broccoli. Sulforaphane induces a battery of detoxification enzymes, including quinone reductase (QR). Induction is thought to be mediated through a common regulatory region termed the antioxidant response element (ARE). To test the hypothesis that the antioxidant selenoprotein thioredoxin reductase (TR) may be induced as part of this coordinated host-defense response to dietary anticarcinogenic compounds, TR activity was measured in livers of rats pair-fed diets containing SF and/or broccoli (n = 6/group). At the doses used, neither SF nor broccoli alone significantly elevated TR activity, whereas treatments containing both broccoli and SF caused a significant increase in TR activity. Glutathione peroxidase (GSH Px), a second selenium-dependant enzyme with antioxidant activity, was downregulated in rats fed both SF and broccoli, compared to the control diet.A second experiment, using mouse hepatoma Hepa1c1c7 cells, tested whether an interaction exists between selenium (Se) and SF in TR inducibility, since Se is known to induce TR activity. Selenium (2.5 &mgr;M) plus SF (2.0 &mgr;M) caused significantly greater TR activity than either treatment alone. All treatments with added Se or SF caused significantly greater TR activities than no Se or SF treatment. Glutathione peroxidase activity was elevated by Se, but not by SF. These data suggest that TR, known to be regulated by Se, is also upregulated as part of a host response to the dietary anticarcinogen SF, a trait not shared by another Se-dependent enzyme, GSH-Px. PMID- 12742545 TI - Effect of resistant starch from corn or rice on glucose control, colonic events, and blood lipid concentrations in streptozotocin-induced diabetic rats. AB - To examine the effect of two types of resistant starch on blood glucose and insulin levels, colonic events, hypolipidemic actions and humoral immune responses, Sprague-Dawley streptozotocin-induced diabetic rats were fed diet containing resistant starch from corn or rice. The marked body weight loss by inducing diabetes was not recovered by feeding resistant starch, even though there are no differences in food intakes compared to the non-diabetic control rats. No significant effect of resistant starch feeding on blood glucose and insulin was found. Even though the length of small intestines, and cecum, colon and rectum together with the tissue weight of cecum were not affected by feeding resistant starch, the intestinal transit time was markedly shortened by both types of resistant starch and resistant starch from corn had a more pronounced effect. The short chain fatty acids in the intestinal contents did not appear to be different among the groups. Nonetheless, both of resistant starch from corn and rice significantly lowered plasma total lipid and cholesterol concentrations compared to the diabetic control. The total liver cholesterol lowering effect was observed with resistant starch from rice. Neither immunoglobulin G nor C(3) were influenced by resistant starch. PMID- 12742547 TI - Dietary 135-fold cholecalciferol supplementation severely disturbs the endochondral ossification in growing dogs. AB - The effects of excessive non-toxic dietary Vitamin D(3) supplementation on Ca homeostasis with specific effects on endochondral ossification and skeletal remodeling were investigated in a group of growing Great Dane dogs supplemented with cholecalciferol (Vitamin D(3); HVitD) versus a control group (CVitD) (1350 microg versus 11.4 microg Vitamin D(3) per kilogram diet) from 6 to 21 weeks of age. There were no differences between groups in plasma concentrations of total Ca, inorganic phosphate, growth hormone, and insulin-like growth factor I and no signs of Vitamin D(3) intoxication in HVitD. For the duration of the study in HVitD compared to CVitD, plasma levels of parathyroid hormone (PTH) decreased, calcitonin (CT) increased, 25-hydroxycholecalciferol [25(OH)D(3)] increased 30- to 75-fold, 24,25-dihydroxycholecalciferol [24,25(OH)(2)D(3)] increased 12- to 16 fold, and 1,25-dihydroxycholecalciferol [1,25(OH)(2)D(3)] decreased by approximately 40%. The latter was attributed to the two-fold increased metabolic clearance rate in the HVitD versus CVitD accompanied by the absence of the anabolic effect of PTH on the production of 1,25(OH)(2)D(3). Fractional Ca absorption (alpha) did not differ between groups at 8 and 14 weeks of age, whereas at 20 weeks of age alpha increased by only 16.4% in HVitD compared to CVitD. Excessive non-toxic Vitamin D(3) supplementation resulted in decreased bone remodeling and focal enlargement of the growth plate with morphology resembling those induced by administration of CT. Hypercalcitoninemia and the imbalanced relationship between 1,25(OH)(2)D(3) and 24,25(OH)(2)D(3) are potent candidates for the disturbed endochondral ossification. PMID- 12742548 TI - Bovine lactoferrin binds to insulin-like growth factor-binding protein-3. AB - Insulin-like growth factor (IGF)-binding protein-3 (IGFBP-3) has been shown to have IGF independent actions that appear to be mediated by specific IGFBP-3 binding proteins located on cell membranes. We show here using Western ligand blotting, a number of mammary membrane proteins that bind 125I-labeled rhIGFBP-3. Immunoprecipitation studies demonstrated that the >70 kDa protein was identified from bovine mammary microsomes as bovine lactoferrin (bLf). In addition to being a secretory protein, Lf is tightly associated with cellular membranes. Labeled rhIGFBP-3 was shown to bind to commercially purchased and processed apo- or holo human or bLf, but not bovine transferrin (bTf). Binding of [125I]rhIGFBP-3 to other positively charged proteins was not detected nor was binding to rhIGFBP-5 or other mammary-secreted IGFBPs observed. Reciprocal specific binding of [125I]bLf to rhIGFBP-3 was shown, but [125I]bTf did not show binding to rhIGFBP 3. While [125I]rhIGF-II does not bind to bLf, unlabeled rhIGF-II was shown to compete with [125I]bLf for rhIGFBP-3 binding. More detailed analysis by dot blot showed that Lf competes (ED(50)=3 microg/ml) or displaces (ED(50)=1mg/ml) bound [125I]rhIGF-II from dot blotted rhIGFBP-3. In vitro studies with a bovine primary mammary epithelial cell culture showed that all-trans-retinoic acid stimulates the appearance of bovine IGFBP-3 and bLf in the conditioned media and that [125I]rhIGFBP-3 could be utilized to detect conditioned media bLf. These findings reveal a novel role for bLf, binding to IGFBP-3 and perhaps disassociating IGFBP 3:IGF when in high concentration. PMID- 12742550 TI - Age-related changes in porcine corticosteroid-binding globulin (pCBG) as determined by an enzyme-linked immunosorbent assay. AB - The objectives of this study were to develop an assay for the direct measure of porcine corticosteroid-binding globulin (pCBG) and to confirm age-related changes in plasma pCBG concentration. Isolation and purification of pCBG from plasma was performed by affinity chromatography and HPLC-DEAE anion exchange techniques. Analysis by SDS-PAGE revealed two polypeptides (54 and 59 kDa) having similar amino acid homology (>50%) to previously reported sequences of seven mammalian species for the first 33 amino acids. Porcine CBG (20 ng/well) was immobilized to microtiter plates and standards or samples added along with rabbit antiserum developed against the purified pCBG. Goat anti-rabbit IgG-alkaline phosphatase conjugate was added followed by p-NPP substrate. The resultant color development was read at 405 nm. Intra- and interassay coefficients of variation (n=26) of a pooled sample were 10 and 15%, respectively. Age-related changes (P<0.001) in plasma pCBG concentration (n=203) from day 3 through 168 of age confirmed that, in the pig, changes seen in the percent distribution of cortisol among protein bound and free forms around day 28 of age are associated with an increase in CBG concentration. PMID- 12742549 TI - Use of a GnRH antagonist, antarelix, associated or not with hCG, to control ovulation in cyclic pony mares. AB - The GnRH antagonist antarelix (Teverelix) was administered to mares (0.01 mg/kg, i.v., twice a day) during the periovulatory period. In Experiment 1, 20 mares were divided into a treated (A3d-) and a control (Control-) group. A3d- mares received antarelix for 3 days from the day when the dominant follicle (F1) reached 32 mm (D0). In Experiment 2, 10 mares were divided into a treated (A6d+) and a control (Control+) group. A6d+ mares received antarelix for 6 days from D0 and hCG was injected in all animals (1600 IU, i.v.) on D1. Pregnancies were determined 13 days after ovulation. In both experiments, antarelix interrupted or totally abolished the LH surge. In Experiment 1, 5/10 of the A3d- mares (with maximum LH concentrations of 11.6 ng/ml at the beginning of treatment) ovulated at the same time as the Control- mares; the other five mares (with LH concentrations under 5.4 ng/ml) ovulated 13.4+/-0.6 days later. In Experiment 2, all the A6d+ mares ovulated at the same time as the Control+ mares. In treated mares which ovulated during the treatment, progesterone concentrations and fertility did not differ from control mares. These results demonstrate that in mares: (1) a small elevation of endogenous LH can induce ovulation, (2) ovulation can be postponed approximately 13 days after a 3-day antarelix treatment if initiated just before the preovulatory LH surge, (3) ovulation can be induced by hCG on depressed levels of endogenous LH, (4) the inhibition of the post ovulatory LH surge has no effect either on the corpus luteum or on fertility. PMID- 12742551 TI - Effect of menhaden fish oil supplementation and lipopolysaccharide exposure on nursery pigs. I. Effects on the immune axis when fed diets containing spray-dried plasma. AB - The objective of the present study was to evaluate the potential immunological benefit of adding menhaden fish oil to the diet of weaned pigs. Twenty-four crossbred male pigs were weaned at approximately 18 days of age and placed on a complex nursery diet containing 30% lactose and 7% plasma protein with 6% corn oil as the fat source (Cont, n=12) or with 5% menhaden fish oil and 1% corn oil as the fat source (MFO, n=12) for a period of 15 days. Body weights did not differ (P>0.78) between dietary groups either at the beginning or end of the 15 days feeding period. On day 15, all pigs were non-surgically fitted with an indwelling jugular catheter. On d 16, pigs received an i.v. injection of either saline (n=6/dietary group) or lipopolysaccharide (LPS; 150 microg/kg body weight; n=6/dietary group) and blood samples were collected at 30 min intervals for a period of 5h. Serum was harvested and stored at -80 degrees C for analysis of cortisol (CS), corticosteroid-binding globulin (CBG), tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma). There was no significant effect of diet on basal concentrations (Time 0) of any of the blood parameters analyzed. A Time x Treatment x Diet interaction (P<0.03) was observed for serum CS such that those pigs which consumed the MFO diet followed by LPS treatment had a reduced CS response as compared to the LPS-treated pigs on the Cont diet. A Time x Treatment interaction (P<0.01) was observed for serum CBG such that LPS treatment reduced circulating CBG as compared to the saline-treated pigs. Time x Treatment x Diet interactions were also observed for serum concentrations of TNF-alpha (P=0.084) and IFN-gamma (P=0.022) such that both the TNF-alpha and IFN-gamma response to the LPS challenge was lower in those pigs receiving the MFO diet as compared to the LPS-treated pigs on the Cont diet. Overall, serum CS was negatively correlated with the CBG response (r=-0.40, P<0.001), however, the strongest negative correlation was observed in the LPS-treated pigs which consumed the MFO diet (r=-0.63, P<0.001). While further studies are needed to evaluate the immunological response of including MFO in the nursery pig diet, the present study demonstrates that supplementation with MFO does indeed alter the immunological response to an LPS challenge. PMID- 12742552 TI - Effect of menhaden fish oil supplementation and lipopolysaccharide exposure on nursery pigs. II. Effects on the immune axis when fed simple or complex diets containing no spray-dried plasma. AB - A trial using 64 weanling pigs (TR4 x PIC C22) was conducted to determine the effects of menhaden fish oil supplementation and diet complexity on performance and immune response of nursery pigs. Pigs (17 days and 6.27+/-1.16 kg) were weaned into a segregated early wean facility and given free access to a complex diet for 7 days post-weaning. At day 0 (day 7 post-weaning), pigs were blocked by weight and allotted to 64 pens. Treatments (Trt) were arranged as a 2 x 2 x 2 factorial arrangement. Main effects included diet (complex versus simple), oil (menhaden fish (MFO) versus corn (CO)), and immunogen (saline versus lipopolysaccharide (LPS)). Experimental diets contained 6% oil (6% CO or 5% MFO+1% CO) and were fed for 14 days. On day 12, i.v. injections of either LPS (150 microg/kg) or saline were given, followed by blood collection at 30 min intervals for 6h. After the immune challenge (day 14), pigs were placed onto a common corn-soybean meal fortified diet and growth performance was evaluated until termination of the study (day 28). Pigs were weighed and feed intakes recorded at 7, 14, and 28 days. Prior to immune challenge (day 12), there were differences in BW for pigs fed complex versus simple diets (P<0.01; 13.1 and 12.1 kg, respectively) and pigs fed CO versus MFO diets (P<0.05; 12.9 and 12.3 kg, respectively). During the challenge period, for pigs treated with LPS there was a Time x Immunogen x Oil effect (P<0.001) for serum cortisol with MFO fed pigs having lower serum cortisol as compared to CO fed pigs. Also, during the challenge period, for pigs treated with LPS there was a Time x Diet x Immunogen x Oil effect (P<0.001) for serum tumor necrosis factor-alpha (TNF-alpha) with pigs fed complex diets supplemented with CO having higher serum TNF-alpha as compared with pigs fed complex diets supplemented with MFO. At days 14 and 28, LPS-treated pigs had lower BW than saline injected controls (P<0.001 and 0.01, respectively). In addition, pigs fed simplified diets continued to have lower BW after challenge compared to pigs fed a complex diet. Interestingly, there were no differences (P>0.10) in BW after challenge in pigs fed MFO. This study suggests that MFO supplementation alters the immune response during LPS challenge and that simplified diets may compromise nursery performance. PMID- 12742553 TI - Cloning and nucleotide sequence of the equine and elk pituitary pre-prolactin cDNA. AB - We report the equine (Equs equs) and elk (Cervus elaphus) pituitary pre-prolactin (PRL) cDNA cloning, and their nucleotide and deduced amino acid sequences. Pre PRL cDNA was obtained by RNA ligation mediated-rapid amplification of cDNA ends (RLM-RACE) and polymerase chain reaction (PCR). The elk pre-PRL cDNA exhibits two polymorphisms at positions 96 and 672, which are silent since they encode for the same amino acids, proline and isoleucine, respectively. We found no polymorphisms in the equine pre-PRL cDNA. The deduced amino acid sequence of the equine pre-PRL is 99% identical to the previously reported protein sequence. Pre-PRL mRNA is <1 kb in length and is highly expressed in the anterior pituitary gland, as demonstrated by Northern hybridization analysis. In summary, we cloned and sequenced the equine and elk pre-PRL cDNAs. The deduced amino acid sequence of elk and equine pre-PRL appears to be moderately conserved among other mammalian species. The polymorphic sites found in the elk cDNA could potentially be used in parentage testing and gene mapping. PMID- 12742554 TI - ARHP editorial. PMID- 12742555 TI - Evaluation of an emergency contraception advance provision service. AB - Timely access to emergency contraception (EC) could increase use when needed, and potentially lead to improved efficacy. We evaluated an advance provision service in the UK. Women were supportive of the service. They indicated that having EC on hand would not change their regular contraceptive use, and supported wider dissemination of information on the service, particularly to younger women. Although some women were supportive of further deregulation of EC, many cited fear of "abuse" or health risks of EC as reasons for strict control. We conclude that advance provision services increase access to EC and are particularly important where EC is not yet available from pharmacists. Providers and women need accurate information on the safety of EC. Uptake of advance provision services could be improved by providing subsidized or free EC to those who cannot pay, and by targeting information to younger women. PMID- 12742556 TI - Efficacy of second versus third generation oral contraceptives in the treatment of hirsutism. AB - OBJECTIVE: To compare second versus third generation combination oral contraceptives (OCs) in the treatment of hirsutism. METHODS: Women with hirsutism, as defined by a minimum Ferriman-Gallwey score of 10, were randomized in a double-blind fashion to receive an OC containing either ethinyl estradiol/desogestrel or ethinyl estradiol/levonorgestrel for 9 months of treatment. Ferriman-Gallwey scores, androgen levels and sex hormone-binding globulin were measured at baseline and every 3 months for the duration of the study. Hormones were measured in duplicate by radioimmunoassay. RESULTS: Of the 47 women enrolled, 24 were randomized to ethinyl estradiol/desogestrel and 23 were randomized to ethinyl estradiol/levonorgestrel. Mean sex hormone-binding globulin increased significantly in subjects using the desogestrel-containing contraceptive compared with the levonorgestrel-containing contraceptive. Ten subjects completed the 9 months of treatment in the levonorgestrel group and 11 completed the study in the desogestrel group. Mean free testosterone and 3alpha androstanediol glucuronide decreased significantly in the group receiving ethinyl estradiol/desogestrel but not in the ethinyl estradiol/levonorgestrel group. Mean Ferriman-Gallwey scores decreased significantly in both treatment groups. Improvement in mean Ferriman-Gallwey score was 35.7 +/- 38.1% (p < 0.001) for the ethinyl estradiol/desogestrel arm and 33.4 +/- 27.3% (p < 0.001) for the ethinyl estradiol/levonorgestrel arm. There were no statistically significant differences found in the improvement of Ferriman-Gallwey scores between the two treatment arms, although the power to detect a difference was limited by the small sample size. CONCLUSIONS: Treatment of hirsute women with third generation OCs containing desogestrel results in a significant increase in sex hormone-binding globulin and decrease in free testosterone and 3alpha-androstanediol glucuronide. Both second and third generation OCs were clinically effective in treating hirsutism. PMID- 12742557 TI - Effects of a low-dose and ultra-low-dose combined oral contraceptive use on bone turnover and bone mineral density in young fertile women: a prospective controlled randomized study. AB - In this prospective, controlled, randomized study, we compared the effect of a low-dose 21-day combined oral contraceptive (COC) containing 20 microg ethinyl estradiol (EE) and 75 microg gestodene (GTD) (Group A; n = 19) with an ultra-low dose 24-day COC containing 15 microg EE and 60 microg GTD (Group B; n = 18) on bone turnover and bone mineral density (BMD) in young, fertile women. Nineteen healthy fertile women were used as untreated controls (Group C). At 3, 6, 9 and 12 months of the study serum osteocalcin (BGP), urinary pyridinoline (PYD) and deoxypyridinoline (D-PYD) were measured in all subjects. At baseline and after 12 months BMD was determined at lumbar spine by dual-energy X-ray absorptiometry in all patients. In both Groups A and B, urinary levels of PYD and D-PYD at 6, 9 and 12 months, were significantly reduced in comparison with basal values and with control subjects (p < 0.05). No significant differences in urinary PYD and D-PYD levels were observed between Groups A and B during the entire period of treatment. At 12 months, no statistically significant difference in spinal BMD values was detected between the three groups and in comparison with basal values. The present study suggests that the two COCs could exert a similar positive effect on bone turnover in young postadolescent women, without any significant and appreciable modification of BMD. PMID- 12742558 TI - Effect of four oral contraceptives on thyroid hormones, adrenal and blood pressure parameters. AB - In a double-blind, controlled, randomized, four-arm, bicentric clinical study, the effect of four oral contraceptives (OCs) on thyroid hormone parameters, cortisol, aldosterone, endothelin-1 and angiotensin II was investigated. Four groups composed of 25 volunteers each (ages between 18 and 35 years) were treated for six cycles with monophasic combinations containing 21 tablets with either 30 microg ethinylestradiol (EE) + 2 mg dienogest (DNG) (30EE/DNG), 20 microg EE + 2 mg DNG (20EE/DNG), 10 microg EE + 2 mg estradiol valerate (EV) + 2 mg DNG (EE/EV/DNG) or 20 microg EE + 100 microg levonorgestrel (LNG) (EE/LNG). The study was completed by 91 subjects. Blood samples were taken by venipuncture after at least 12 h fasting on Day 21-26 of the control cycle and on Day 18-21 of the first, third and sixth treatment cycle. There was a significant increase in triiodothyronine (T3) and thyroxine (T4) by 20-40% in all treatment cycles, while thyroid-stimulating hormone was significantly increased only with EE/EV/DNG. Treatment with the DNG-containing OCs caused no change in free T4 (FT4) and a transitory reduction in free T3 (FT3) levels during the first cycle. During intake of EE/LNG, FT4 rose slightly, while FT3 was not altered. The pronounced rise in the serum concentrations of cortisol appeared to be related to the EE dose. During the first three cycles of treatment, no effect on angiotensin II levels was observed, while in the sixth cycle a significant decrease was measured in all treatment groups. The four OCs did not influence the serum concentrations of endothelin-1 and no consistent effects were found concerning those of aldosterone. The results suggest that the three DNG-containing and the LNG containing low-dose OCs may increase T3, T4 and cortisol due to an elevated binding to serum globulins, while the free proportion of the hormones is not or only slightly changed. Therefore, these OCs have only minor effects on thyroid function, adrenal and blood pressure serum parameters. PMID- 12742559 TI - A 13-month multicenter clinical experience of a low-dose monophasic oral contraceptive containing 20 microg ethinylestradiol and 75 microg gestodene in Latin American women. AB - This prospective, multicenter study was conducted to evaluate the contraceptive reliability, cycle control and tolerability of a 21-day oral contraceptive regimen containing 20 microg ethinylestradiol and 75 microg gestodene in four Latin American countries (Mexico, Argentina, Brazil and Colombia). Participants took trial medication daily for 21 days. Contraceptive efficacy, cycle control and tolerability were evaluated over a period of 13 cycles. Efficacy data gathered from 5,109 treatment cycles were obtained from 393 participants. The trial medication proved to be an effective contraceptive and provided good cycle control. One pregnancy because of poor compliance was recorded. This resulted in a study Pearl index of 0.25. Forty-six percent of Latin American women reported one intracyclic spotting bleeding episode and 37.6% reported one intracyclic breakthrough bleeding (medium/excessive bleeding) episode during cycles 2-4 (primary target). Overall, intracyclic bleeding was reported in 41%. Overall, there was a trend towards a lower incidence of spotting in all the countries and this difference had statistical significance between Argentina and the others three countries (p < 0.05) during cycles 2-4. This trend was also apparent with respect to breakthrough bleeding, but again the difference did not achieve statistical significance. The discontinuation rate because of adverse events was low (3%); no serious adverse events were reported. More than 78% of the women in the four countries maintained constant body weight or lost weight (2 kg) during the study. The treatment effect on blood pressure was negligible. There were no appreciable changes in mean laboratory values over the course of the study. PMID- 12742560 TI - Characteristics of menstrual problems associated with Norplant discontinuation: results of a multinational study. AB - This paper presents a secondary analysis of 3419 Norplant users from 11 countries throughout Asia, Africa and Latin America. The main objective was to identify the menstrual problems associated with a high risk of early discontinuation of Norplant use. Eligible participants were divided into two groups: those who continued use (n = 2667) for the first 3 years and those who discontinued use because of bleeding or pain (n = 752) during the first 3 years of use. Menstrual characteristics of the women were compared between groups using a logistic regression model with generalized estimating equations. Flow duration of more than 7 days, excessive amount of flow, dysmenorrhea and intermenstrual bleeding for more than 7 days were associated with a significant risk of discontinuation. The highest risk was observed in association with flow duration of more than 7 days (adjusted odds ratios of 1.85, 2.01 and 2.20 at 1, 2 and 3 years, respectively). The appropriate management of prolonged or excessive bleeding, namely by continuous counseling, is essential for successful Norplant use. PMID- 12742561 TI - Survey of IUD replacing status in Sichuan, PR China. AB - To investigate the status of copper-bearing intrauterine device (IUD) replacing inert IUD in Sichuan Province, cluster random sampling method was adopted for the survey. Questionnaires were completed and IUD were inspected through ultrasound among 12,804 subjects of 15 counties, who were inserted an IUD from 1994 to 1998. The insertion rate of copper-bearing IUD was 42.1%, in a gradually increasing trend; it was lower in mountain areas than in plain and hilly areas and increased after induced abortion and previous IUD failure. Some factors, such as lower pregnancy rate, lower expulsion rate and easy removal, etc., contributed to copper-bearing IUD use. On the other hand, other factors like longer contraception period, lower side effects and lower cost contributed to inert IUD use. The results show that the adoption of copper-bearing IUD is increasing, but those factors influencing copper-bearing IUD replacement should not be neglected. PMID- 12742562 TI - Frustrated demand for postpartum female sterilization in Brazil. AB - During the last three decades, Brazilians have relied almost exclusively on two contraceptive methods, the pill and female sterilization, with sterilization use increasing over time. Until a new law was passed in 1997, sterilization was virtually illegal and not covered by either public or private health insurance. It was, however, frequently provided in public and private hospitals in conjunction with a cesarean section. The new law regulating sterilization provided for reimbursement for interval sterilizations by public health insurance, but placed restrictions on availability intended to reduce the use of cesareans. These restrictions included the prohibition of postpartum sterilizations. This paper focuses on women's sterilization intentions during pregnancy and their experiences postpartum. In a prospective study of 1612 pregnant women carried out in four Brazilian cities, there was substantial demand for postpartum sterilization in both the private and public sectors among women who wanted no more children. However, public patients were much less likely to be sterilized than private patients. Thus, the new law may not have reduced inequities in access or, paradoxically, the incentive for unnecessary cesarean sections. PMID- 12742563 TI - Women's preferences for vaginal antimicrobial contraceptives. V: attitudes of Brazilian women to the insertion of vaginal products. AB - The rapid spread of HIV/AIDS in the female population increases the urgency of developing new formulations that offer protection from this disease as well as other sexually transmitted infections. In many cultures, women do not readily accept touching their genitals or inserting products into their vaginas. Information on this subject was collected during a study involving 635 women in Brazil to determine the preferred attributes of vaginal products. Seventy-six percent would use an idealized contraceptive method that offered dual protection even though it could only be inserted with a finger and 96% would use this method if it could only be placed with an applicator. Qualitative analyses of responses to open questions suggest that the majority of Brazilian women studied did not like to touch their vagina with their finger or to insert devices. Although the introduction of safe and effective vaginal microbicides into many cultural settings can be successful, it should be accompanied by significant efforts to educate women about their bodies. PMID- 12742564 TI - Vasectomy effectiveness in Nepal: a retrospective study. AB - The main purpose of this retrospective, cross-sectional study was to evaluate the effectiveness of vasectomy in an ongoing public sector program in Nepal. We evaluated semen samples from men who had previously had a vasectomy, and asked about the occurrence of pregnancies in the men's partners. In addition, the surgeons who performed the vasectomies completed a questionnaire about their techniques. A two-stage stratified sampling procedure was used to select 1263 men from among over 30,000 men, who had previously undergone a no-scalpel vasectomy, mostly by ligation and excision, in 32 districts between July 1996 and June 1999. Semen samples were preserved and analyzed at a central laboratory. A US andrology laboratory validated the lab results. Twenty-three men (2.3%, 95% confidence interval [CI] 1.1-3.6) had >/=500,000 sperm/mL in their semen. Fifteen of those men reported pregnancies conceived after their vasectomy. In addition, six men with azoospermia reported pregnancies for which conception occurred within 3 months after vasectomy. Eleven men with azoospermia reported pregnancies for which conception occurred more than 3 months after vasectomy. Reported pregnancy was more likely in younger partners. The life table pregnancy rates for all men interviewed were 0.7 (95% CI 0.2-1.1), 1.7 (95% CI 1.4-2.1) and 4.2% (95% CI 3.2 5.2) at 3, 12 and 36 months, respectively. In low-resource, programmatic settings, vasectomy failure rates may be higher than commonly cited rates, especially in younger populations. Additional research is needed to determine if other occlusion techniques could reduce failure rates. Counseling on vasectomy should always convey the possibility of failure and partner pregnancy. PMID- 12742565 TI - Mechanism of action of some acrylophenones, quinolines and dithiocarbamate as potent, non-detergent spermicidal agents. AB - Some suitably substituted acrylophenones, quinolines and dithiocarbamate were synthesized as new generation, non-detergent spermicides and were studied for their mechanism of action in comparison with various known spermicides belonging to several different classes of chemical compound. Nonoxynol-9, benzalkonium chloride, Sapindus saponins, verapamil, emetine and tartaric acid were used as reference molecules to study the effect of new spermicides on human sperm motility parameters (using computer-assisted semen analyzer), plasma membrane integrity, lipid peroxidation and defense system against reactive oxygen species (ROS). Results have indicated that sperm plasma membrane remains the primary site of action of most of the spermicides, though the effect may be predominantly on the physiological integrity rather than the structural integrity in case of the new compounds. Lipid peroxidation may play an important role in disrupting sperm membrane physiology that may or may not be accompanied with a detrimental effect on the defense system of the human spermatozoa against the ROS. PMID- 12742566 TI - Effect of oral tamoxifen on semen characteristics and serum hormone profile in male bonnet monkeys. AB - The effects of oral tamoxifen were studied at a dose of 0.4 mg/kg per day, on the serum hormones and semen parameters in adult male bonnet monkeys, for a period of 90 days. Honey was used as vehicle. Monkeys were treated with honey for 30 days, followed by tamoxifen from Day 30-120 (90 days). Thereafter the treatment was withdrawn until Day 150 of schedule. Blood samples were drawn at 12 and 24 clock hours at monthly intervals for the analysis of luteinizing hormone, follicle stimulating hormone and testosterone. Semen samples were also collected for analysis once a month, from Day 0-150 of exposure. Tamoxifen treatment produced a transient but significant increase in circulating gonadotropins, at Day 90 of treatment schedule, corresponding to 60 days of treatment. Whilst serum testosterone levels were normal throughout treatment period, an increase was observed after 30 days of drug withdrawal. No effect of oral tamoxifen was evident on semen parameters, viz., volume, counts, morphology and motility. However, throughout the exposure period to honey, a significant increase was observed in sperm counts without any effect on testosterone levels. The present study suggests that oral tamoxifen has a transient antiestrogenic effect on the serum hormones and no effect on semen parameters of adult nonhuman primate males. It is concluded that bioefficacy of oral tamoxifen may have been reduced due to hepatic metabolism. PMID- 12742567 TI - Postcoital treatment with levonorgestrel does not disrupt postfertilization events in the rat. AB - Levonorgestrel (LNG), a progestin widely used for regular hormonal contraception, is also used for emergency contraception (EC) to prevent pregnancy after unprotected intercourse. However, its mode of action in EC is only partially understood. One unresolved question is whether or not EC prevents pregnancy by interfering with postfertilization events. Here, we report the effects of acute treatment with LNG upon ovulation, fertilization and implantation in the rat. LNG inhibited ovulation totally or partially, depending on the timing of treatment and/or total dose administered, whereas it had no effect on fertilization or implantation when it was administered shortly before or after mating, or before implantation. It is concluded that acute postcoital administration of LNG at doses several-fold higher than those used for EC in women, which are able to inhibit ovulation, had no postfertilization effect that impairs fertility in the rat. PMID- 12742568 TI - Commentary on repeat emergency contraception. PMID- 12742570 TI - Pathogenesis and potential antiviral therapy of complications of smallpox vaccination. AB - Vaccination against smallpox may result in a variety of complications, ranging in severity from benign to lethal. Universal vaccination was halted in the US in 1972, so almost half the present population has never been vaccinated. Because side effects occur most often in first-time vaccinees, current plans for rapid large-scale vaccination in the event of bioterrorist attack raise concerns about the occurrence of a large number of adverse events. Most complications result from the excessive replication of vaccinia virus, making them potential targets for antiviral therapy. Effective treatment is especially needed for persons with atopic dermatitis or eczema, who are unusually susceptible to the initiation and spread of vaccinia infection because of defects of innate immunity in the skin, and for individuals with defective cell-mediated immunity, who are unable to eliminate vaccinia infection once it has begun. In the past, many complications were treated with vaccinia immune globulin (VIG) and/or the antiviral drug methisazone, but neither was tested in placebo-controlled trials. New antiviral drugs are now available, but have not yet been evaluated for treating vaccinia infections in humans. Both laboratory research and clinical studies are needed to help prevent serious complications in any major vaccination campaign. PMID- 12742572 TI - A new sensitive cartridge-RIA method for determination of stavudine (D4T) triphosphate in human cells in vivo. AB - We describe a simple and sensitive method to determine stavudine triphosphate, the active intracellular anabolite of stavudine (D4T). Quantification of D4T triphosphate was performed with a combined cartridge-radioimmunoassay (cartridge RIA) which enabled us to measure concentrations of D4T triphosphate as low as 0.5 ng/ml, or an intracellular concentration which corresponds to 20 fmol/10(6) cells if diluted like our previously published zidovudine (ZDV) assay. The only alternate methodology at present employs liquid chromatography mass spectroscopy (LC-MS/MS). The use of the cartridge-RIA methodology provides a cost-effective alternative for the determination of in vivo cellular pharmacokinetics studies of D4T in human immunodeficiency virus (HIV)-infected persons. PMID- 12742571 TI - The compound DATEM inhibits respiratory syncytial virus fusion activity with epithelial cells. AB - The effect of diacetyltartaric acid esters of mono and diglycerides (DATEM) on fusion of respiratory syncytial virus (RSV) with HEp-2 cells was studied using the R18 fluorescence dequenching fusion assay. At DATEM concentrations less than 2.0 microg/ml, the inhibition of fusion increased with the concentration of DATEM. At 2 microg/ml of DATEM, the fusion was suppressed by 80-90%. Studies examining possible mechanism of fusion-inhibition indicated that DATEM was likely adsorbed onto lipid membranes of both viral envelope and target cell membranes. Quantitative measurements of DATEM adsorption onto membranes were also performed using lipid monolayers and vesicles. The surface pressure of lipid monolayer formed at the air/aqueous interface increased as the concentration of DATEM in the monolayer subphase increased, suggesting that DATEM was inserted into the monolayer. As the concentration of DATEM in vesicle suspensions increased, electrophoretic mobility of initially uncharged lipid vesicles also increased, reflective of increased negative charge at vesicle surfaces. These results strongly suggest that the insertion of DATEM onto membranes inhibited viral fusion. DATEM may prove to be effective in limiting the infectivity of RSV by interference with the fusion of the viral envelope with target cell membranes. PMID- 12742573 TI - Direct antiviral effect of cycloferon (10-carboxymethyl-9-acridanone) against adenovirus type 6 in vitro. AB - Adenoviruses represent a broad group of human pathogens that currently have no specific and safe drugs for treatment. We demonstrated direct (non IFN-mediated) antiviral activity of cycloferon (10-carboxymethyl-9-acridanone, CMA), a potent interferon inducer, against adenovirus type 6 (Ad6) in Hep-2 cells. Virus production and details of morphogenesis were studied by ELISA with antibodies to the Ad6 hexon protein, and transmission electron microscopy, respectively. Immunoenzyme assay revealed that CMA does not inhibit viral protein synthesis but instead strongly reduces the ability of the virus to generate infectious progeny virus in a dose dependent manner. Ultrastructural study shows that CMA alters the structure of intranuclear virus-specific inclusions. We suggest that CMA suppresses the late stages of viral cycle in the infected cell. PMID- 12742574 TI - Susceptibility to highly sulphated glycosaminoglycans of human immunodeficiency virus type 1 replication in peripheral blood lymphocytes and monocyte-derived macrophages cell cultures. AB - In the search for new drugs against human immunodeficiency virus type 1 (HIV-1), the replication of III(B) and BaL strains, and of seven primary isolates from AIDS patients, cultured both in peripheral blood lymphocytes (PBLs) and in monocyte-derived macrophages (MACs), was investigated in the presence of two dermatan sulphate and heparin at 10 microg/ml. The three polysaccharides effectively inhibited the replication of III(B) in PBLs and of BaL in MACs, while producing either a slight inhibition or an unexpected large increase in the replication of the seven primary isolates, especially in MAC cultures. In one case, stimulation was found in PBLs and, at lower doses, also with BaL in MACs. Co-receptor use, adaptation to C8166 T cell line, partial sequence of the gp120 V3 loop, variation in positive charge distribution and number of potential glycosylation sites along the V3 loop were assessed for each strain. No explanation could be found for the different susceptibility of the viruses to the polysaccharides. Their presence probably brings about both inhibitory and stimulatory effects, the final outcome depending on the virus, cells and polysaccharide. PMID- 12742575 TI - Determination of antiviral efficacy against lymphotropic herpesviruses utilizing flow cytometry. AB - Epstein-Barr virus (EBV), human herpesvirus type 6 (HHV-6), and human herpesvirus type 8 (HHV-8) comprise a group of lymphotropic herpesviruses which are responsible for a wide range of diseases, including lymphoproliferative disorders and tumors. We have developed several flow cytometric assay (FACS) systems to evaluate antiviral efficacy against EBV, HHV-6 and HHV-8. Assays using either EBV or HHV-8, members of the gammaherpesvirus subfamily, have shown that while EBV responds well to acyclovir (ACV), HHV-8 was most sensitive to cidofovir (CDV). Since HHV-6 strains are divided into two sub-groups, A and B, we evaluated antiviral efficacy for strains from each group. The group A strain, HHV-6(GS), was inhibited by foscarnet (PFA), CDV and ganciclovir (GCV) in both Sup-T1 and HSB-2 cell lines. HHV-6(Z-29), a representative group B virus, was inhibited by GCV and CDV but not by PFA. Our findings indicate that flow cytometry can be utilized to efficiently evaluate new antiviral agents against lymphotropic herpesviruses and that the results are comparable to those obtained by other methods such as immunofluorescence. PMID- 12742576 TI - Synergy of bovine lactoferrin with the anti-cytomegalovirus drug cidofovir in vitro. AB - Human cytomegalovirus (HCMV) causes severe morbidity and mortality in immunocompromised patients. Treatment of HCMV infections with conventional antiviral drugs like ganciclovir and cidofovir has major drawbacks (i.e. serious side effects). Therefore, combination therapies using drugs with different antiviral mechanisms should be envisaged. Potential synergy between lactoferrin (LF), an antibacterial, antimycotic and antiviral protein, and the antiviral drugs acyclovir, ganciclovir, foscarnet and cidofovir was investigated, using an in vitro test system with the recombinant RC256 HCMV strain. RESULTS: Combination of LF with acyclovir and foscarnet resulted in antagonism. When LF and ganciclovir were combined, neither synergy nor antagonism was observed. Strikingly, the combination of LF with cidofovir resulted in marked synergy. The synergistic effect could be explained by inhibition of two subsequent steps in the viral replication cycle: HCMV penetration into the target cells and intracellular synthesis of HCMV DNA. In conclusion, LF might be a potential candidate for combination therapy with cidofovir. PMID- 12742577 TI - Inhibition of adenovirus infection and adenain by green tea catechins. AB - Green tea catechins have been reported to inhibit proteases involved in cancer metastasis and infection by influenza virus and HIV. To date there are no effective anti-adenoviral therapies. Consequently, we studied the effect of green tea catechins, and particularly the predominant component, epigallocatechin-3 gallate (EGCG), on adenovirus infection and the viral protease adenain, in cell culture. Adding EGCG (100 microM) to the medium of infected cells reduced virus yield by two orders of magnitude, giving and IC(50) of 25 microM and a therapeutic index of 22 in Hep2 cells. The agent was the most effective when added to the cells during the transition from the early to the late phase of viral infection suggesting that EGCG inhibits one or more late steps in virus infection. One of these steps appears to be virus assembly because the titer of infectious virus and the production of physical particles was much more affected than the synthesis of virus proteins. Another step might be the maturation cleavages carried out by adenain. Of the four catechins tested on adenain, EGCG was the most inhibitory with an IC(50) of 109 microM, compared with an IC(50) of 714 microM for PCMB, a standard cysteine protease inhibitor. EGCG and different green teas inactivated purified adenovirions with IC(50) of 250 and 245-3095, respectively. We conclude that the anti-adenoviral activity of EGCG manifests itself through several mechanisms, both outside and inside the cell, but at effective drug concentrations well above that reported in the serum of green tea drinkers. PMID- 12742578 TI - Inhibition of wild-type human immunodeficiency virus and reverse transcriptase inhibitor-resistant variants by Phyllanthus amarus. AB - Substantial progress has been made in research on natural products which effectively inhibit HIV-1 replication. Many active compounds were isolated from traditionally used medicinal plants including Phyllanthus species. This study shows that aqueous as well as alcohol-based Phyllanthus amarus extracts potently inhibit HIV-1 replication in HeLa CD4(+) cells with 50% effective concentration (EC(50)) values ranging from 0.9 to 7.6 microg/ml. A gallotannin enriched fraction showed enhanced activity (0.4 microg/ml), and the purified gallotannins geraniin and corilagin were most active (0.24 microg/ml). HIV-1 replication was also blocked in CD4(+) lymphoid cells with comparable EC(50) values. Applying a cell-based internalization assay, we could demonstrate 70-75% inhibition of virus uptake at concentrations of 2.5 microg/ml for the water/alcohol extract and geraniin. In addition, a concentration-dependent inhibition of HIV-1 reverse transcriptase (RT) could be demonstrated in vitro. The 50% inhibitory concentration (IC(50)) values varied from 1.8 to 14.6 microg/ml. The ability to inhibit replication of a variety of RT inhibitor-resistant HIV-1 strains points to the potential of P. amarus extracts, as natural products, in the chemotherapy of HIV infections. PMID- 12742579 TI - Downregulation of Trypanosoma brucei VSG expression site promoters on circular bacterial artificial chromosomes. AB - Trypanosoma brucei has about 20 telomeric variant surface glycoprotein (VSG) gene expression sites (ESs), which are downregulated in the insect form. We investigated the transcriptional behaviour of ES promoters on bacterial artificial chromosomes (BACs) containing two different ESs and their flanking regions on fragments of about 140kb. Four different BACs containing either the 221 or the VO2 ES were introduced into insect form T. brucei. The BACs replicated as circular episomes as shown using pulsed field gel (PFG) analysis of DNA exposed to increasing doses of gamma radiation, and digestion with Dam methylation-sensitive restriction enzymes. BAC copy number per cell varied from about 3 for the 221 ES BACs to about 15 for the VO2 ES BACs. Increasing drug selection pressure on the VO2 BAC T. brucei transformants resulted in amplification to about 80 BACs per cell. Although BACs were maintained in the absence of drug selection for at least 56 days, copy number fell and there was no evidence for centromere activity. ES promoters on small plasmid episomes introduced into insect form T. brucei in transient transfections are derepressed. In contrast, ES promoters on large BAC episomes are downregulated both on the original ES BACs, and on ES BACs selected for a drug marker driven by a rDNA promoter fused to the BAC vector. This indicates that downregulation of ES promoters in insect form T. brucei is influenced by genomic context, but does not necessitate proximity to a chromosome end. PMID- 12742580 TI - Structural features affecting variant surface glycoprotein expression in Trypanosoma brucei. AB - The glycosylphosphatidylinositol (GPI)-anchored variant surface glycoprotein (VSG) of Trypanosoma brucei is the most abundant GPI-anchored protein expressed on any cell, and is an essential virulence factor. To determine what structural features affect efficient expression of VSG, we made a series of mutations in two VSGs. Inserting 18 amino acids, between the amino- and carboxy-terminal domains, reduced the expression of VSG 221 to about 3% of the wild-type level. When this insertion was combined with deletion of the single carboxy-terminal subdomain, expression was reduced a further three-fold. In VSG 117, which contains two carboxy-terminal subdomains, point mutation of the intervening N-glycosylation site reduced expression about 15-fold. Deleting the most carboxy-terminal subdomain and intervening region, including the N-glycosylation site, reduced expression to 15-20% of wild type VSG, and deletion of both subdomains reduced expression to <1%. Despite their low abundance, all VSG mutants were GPI anchored on the cell surface. Our results suggest that, for a protein to be efficiently displayed on the surface of bloodstream-form T. brucei, it is essential that it contains the conserved structural motifs of a T. brucei VSG. Serum resistance associated protein (SRA), which confers human infectivity on T. brucei, strongly resembles a VSG deletion mutant. Expression of three epitope-tagged versions of SRA in T. brucei conferred total resistance to human serum. SRA possesses a canonical GPI signal sequence, but we were unable to obtain unequivocal evidence for the presence of a GPI anchor. SRA was not released during osmotic lysis, indicating that it is not GPI anchored on the cell surface. PMID- 12742581 TI - Delineation of the regulated Variant Surface Glycoprotein gene expression site domain of Trypanosoma brucei. AB - The African trypanosome Trypanosoma brucei is protected in the bloodstream of the mammalian host by a dense Variant Surface Glycoprotein (VSG) coat. Although an individual cell has hundreds of VSG genes, the active VSG is transcribed in a mutually exclusive fashion from one of about twenty telomeric VSG expression sites. Expression sites are regulated domains flanked by 50 bp repeat arrays and extensive tracts of repetitive elements. We have integrated exogenous rDNA and expression site promoters upstream of the 50 bp repeats of the VO2 VSG expression site. Transcription from both types of exogenous promoter is downregulated and comparable to promoters targeted into the VSG Basic Copy arrays. We show that the upstream exogenous rDNA promoter escapes VSG expression site control, as switching the downstream VO2 VSG expression site on and off does not affect its activity. Therefore, the 50 bp repeat arrays appear to be the boundary of the regulated expression site domain. PMID- 12742583 TI - Identification of an encystation-specific transcription factor, Myb protein in Giardia lamblia. AB - The life cycle of Giardia lamblia contains two differentiation processes, encystation and excystation. We performed an experiment to identify the genes induced during encystation using the differential display reverse transcriptase polymerase chain reaction. Three of twelve isolated cDNA clones that showed increased transcription during encystation were identified to be of the myb2, which encodes a well-known transcriptional factor involved in cellular development and differentiation. The amino acid sequences of the Myb2 protein deduced from the isolated gene revealed that this Myb2 has a DNA binding domain comprising two imperfect repeats at its carboxyl-terminus. The nuclear localization of Myb2 protein during encystation was observed in vivo by expressing a Myb2-GFP fusion protein. In a random site selection experiment, the oligonucleotides bound by rMyb2 contained a conserved sequence of GTTT(G/T)(G/T). Two promoters of the encystation-induced genes, myb2, and cwp1, were also found to bind to rMyb2, whereas gap1, one of the constitutive genes did not. PMID- 12742582 TI - Plasmodium falciparum histidine-rich protein II binds to actin, phosphatidylinositol 4,5-bisphosphate and erythrocyte ghosts in a pH-dependent manner and undergoes coil-to-helix transitions in anionic micelles. AB - The recombinant histidine-rich protein II (HRPII) from Plasmodium falciparum was shown to bind actin and phosphatidylinositol 4,5-bisphosphate (PIP(2)) in vitro in a pH-dependent manner, very similar to hisactophilin, an actin-binding protein from ameba. Binding of HRPII to actin and PIP(2) occurred at pH 6.0 and 6.5, but not above pH 7.0. Circular dichroism (CD) spectroscopy confirmed that HRPII interacts with actin at pH below 7.0, as judged by the changes induced in the secondary structure of the HRPII/actin mixture. Further CD analysis demonstrated that HRPII adopts a predominantly alpha-helical conformation with anionic micelles of PIP(2) and SDS, but not with neutral micelles of phosphatidylcholine (PC), a feature that is common to many actin-binding proteins involved in cytoskeleton remodeling. Similarly to hisactophilin, a GFP-HRPII fusion protein shuttled from the cytoplasm to the nucleus of HeLa cells as the cellular pH was lowered from 8.0 to 6.0. HeLa cells transfected with the HRPII gene showed increased levels of histidine-rich proteins (HRPs) in the soluble cell fraction at pH 8.0. At pH 6.0, however, HRPs were detected mainly in the insoluble cell fraction. Interestingly, we found that HRPII binds to human erythrocyte membranes at pH 6.0 and 6.5 but not at pH above 7.0. Our results point to remarkable similarities between HRPII, hisactophilin, and actin-binding proteins. Possible roles of the HRPII during Plasmodium infection are discussed in the light of these findings. PMID- 12742584 TI - Isolation and characterization of the stage-specific cytochrome b small subunit (CybS) of Ascaris suum complex II from the aerobic respiratory chain of larval mitochondria. AB - We recently reported that Ascaris suum mitochondria express stage-specific isoforms of complex II: the flavoprotein subunit and the small subunit of cytochrome b (CybS) of the larval complex II differ from those of adult enzyme, while two complex IIs share a common iron-sulfur cluster subunit (Ip). In the present study, A. suum larval complex II was highly purified to characterize the larval cytochrome b subunits in more detail. Peptide mass fingerprinting and N terminal amino acid sequencing showed that the larval and adult cytochrome b (CybL) proteins are identical. In contrast, cDNA sequences revealed that the small subunit of larval cytochrome b (CybS(L)) is distinct from the adult CybS (CybS(A)). Furthermore, Northern analysis and immunoblotting showed stage specific expression of CybS(L) and CybS(A) in larval and adult mitochondria, respectively. Enzymatic assays revealed that the ratio of rhodoquinol-fumarate reductase (RQFR) to succinate-ubiquinone reductase (SQR) activities and the K(m) values for quinones are almost identical for the adult and larval complex IIs, but that the fumarate reductase (FRD) activity is higher for the adult form than for the larval form. These results indicate that the adult and larval A. suum complex IIs have different properties than the complex II of the mammalian host and that the larval complex II is able to function as a RQFR. Such RQFR activity of the larval complex II would be essential for rapid adaptation to the dramatic change of oxygen availability during infection of the host. PMID- 12742585 TI - ICAM-1 can play a major role in mediating P. falciparum adhesion to endothelium under flow. AB - We have investigated the importance of adhesion molecule co-operation in mediating Plasmodium falciparum adhesion to endothelial cells under flow conditions. Using three laboratory parasite lines and a patient isolate which differ in their ICAM-1 and CD36-binding avidity, we found that blockade of ICAM-1 and CD36 separately reduced IRBC adhesion by up to 95 and 50%, respectively. These results confirm previous data showing that ICAM-1 and CD36 synergize to mediate adhesion, but differ in demonstrating that without ICAM-1, binding under flow conditions is severely impaired. Thus, in this system, ICAM-1 is critical for P. falciparum adhesion to activated endothelium and once bound, synergy with CD36 mediates the majority (> or =98%) of adhesion. PMID- 12742586 TI - Development and pre-clinical analysis of a Plasmodium falciparum Merozoite Surface Protein-1(42) malaria vaccine. AB - Merozoite Surface Protein-1(42) (MSP-1(42)) is a leading vaccine candidate against erythrocytic malaria parasites. We cloned and expressed Plasmodium falciparum MSP-1(42) (3D7 clone) in Escherichia coli. The antigen was purified to greater than 95% homogeneity by using nickel-, Q- and carboxy-methyl (CM) substituted resins. The final product, designated Falciparum Merozoite Protein-1 (FMP1), had endotoxin levels significantly lower than FDA standards. It was structurally correct based on binding conformation-dependent mAbs, and was stable. Functional antibodies from rabbits vaccinated with FMP1 in Freund's adjuvant inhibited parasite growth in vitro and also inhibited secondary processing of MSP-1(42). FMP1 formulated with GlaxoSmithKline Biologicals (GSK) adjuvant, AS02A or alum was safe and immunogenic in rhesus (Macaca mulatta) monkeys. PMID- 12742587 TI - Using RNA interference to manipulate endogenous gene expression in Schistosoma mansoni sporocysts. AB - Direct assessments of gene function in parasitic flatworms have been hampered by the lack of effective tools to alter gene expression. The aim of the present study was to use RNA-interference (RNAi) to achieve targeted gene knockdown in larval stages of the human blood fluke, Schistosoma mansoni. We selected two S. mansoni genes for RNAi experiments: SGTP1, a facilitated diffusion glucose transporter and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). When S. mansoni larvae were treated in vitro for 6 days with dsRNA specific to one of these two genes, targeted transcript levels were reduced by 70-80% as determined by quantitative PCR (qPCR), while non-targeted transcripts were unaffected. Parasite exposure to SGTP1 dsRNA, but not GAPDH dsRNA, reduced larval glucose-uptake capacity by 40%, demonstrating that SGTP1 transcript knockdown results in the functional phenotype of reduced glucose transport activity. The effect of dsRNA treatment on transcript level was evident for up to 28 days after an initial dsRNA treatment. Interestingly, dsRNA treatment was effective only when miracidia were allowed to undergo the transition to sporocysts in its presence, while treatment of fully transformed sporocysts was ineffective. Fluorescence patterns in larvae exposed to rhodamine-labeled dsRNA as miracidia and sporocysts were similar, suggesting that the difference in susceptibility to dsRNA treatment between the two life stages may not be due to differences in dsRNA entry. Overall, this technology will enable direct assessment of the roles of individual genes in physiological processes of larval stages of S. mansoni, a crucial step in the identification of novel intervention targets for this important human pathogen. PMID- 12742588 TI - Improvements in transfection efficiency and tests of RNA interference (RNAi) approaches in the protozoan parasite Leishmania. AB - Approaches which eliminate mRNA expression directly are ideally suited for reverse genetics applications in eukaryotic microbes which are asexual diploids, such as the protozoan parasite Leishmania. RNA interference (RNAi) approaches have been successful in many species, including the related parasite Trypanosoma brucei. For RNAi tests in Leishmania, we developed improved protocols for transient and stable DNA transfection, attaining efficiencies of up to 25 and 3%, respectively. This facilitated RNAi tests at the alpha-tubulin locus, whose inhibition gives a strong lethal phenotype in trypanosomatids. However, transient or stable transfection of DNAs encoding mRNAs for an alpha-tubulin stem-loop construct and GFP to monitor transfection resulted in no effect on parasite morphology, growth or tubulin expression in Leishmania major or L. donovani. Transient transfection of a 24-nucleotide double-stranded alpha-tubulin siRNA also had no effect. Similar results were obtained in studies targeting an introduced GFP gene with a GFP stem-loop construct. These data suggest that typical RNAi strategies may not work effectively in Leishmania, and raise the possibility that Leishmania is naturally deficient for RNAi activity, like Saccharomyces cerevisae. The implications to parasite biology, gene amplification, and genetic analysis are discussed. PMID- 12742589 TI - Comparison between constitutive and inducible plasmid vectors used for gene expression in Entamoeba histolytica. PMID- 12742590 TI - Long-term functional consequences of transient occlusion of the middle cerebral artery in rats: a 1-year follow-up of the development of epileptogenesis and memory impairment in relation to sensorimotor deficits. AB - Post-stroke seizures occur in 5-20% of patients. Modeling of stroke-induced seizures in animals provides a useful tool for investigating the molecular basis of epileptogenesis and for developing therapies for stroke patients at increased risk for epileptogenesis. The questions addressed in the study were: (1) Do rats develop spontaneous seizures after transient occlusion of the middle cerebral artery (MCAO)? (2) Is epileptogenesis associated with impaired hippocampus dependent spatial learning and memory? (3) Are the functional abnormalities linked to axonal plasticity in the dentate gyrus? (4) Does the sensorimotor impairment induced by MCAO predict the risk of epileptogenesis? Adult male Sprague-Dawley rats were subjected to MCAO for 120 min. Development of spontaneous seizures was monitored by 1 week of continuous video electroencephalographic (EEG) recordings at 3, 7, and 12 months after MCAO. Spontaneous seizures were not detected during 1-year follow-up in ischemic rats. Animals were, however, impaired in the spatial memory task (P<0.001), which was not associated with altered hippocampal LTP or abnormal mossy fiber sprouting (Timm staining). Animals also had a long-lasting sensorimotor deficit (P<0.05). The present study indicates that MCAO causes long-lasting sensorimotor and spatial memory impairment, but does not induce epileptogenesis or spontaneous seizures. PMID- 12742591 TI - Allopregnanolone serum levels and expression of 5 alpha-reductase and 3 alpha hydroxysteroid dehydrogenase isoforms in hippocampal and temporal cortex of patients with epilepsy. AB - In the human central nervous system, progesterone is rapidly metabolised to 5 alpha-dihydroprogesterone which subsequently is further reduced to allopregnanolone (AP). These conversions are catalysed by 5 alpha-reductase and 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSD). Although different isoforms of both enzymes have been identified in the brain, our knowledge of their expression in the human brain remains limited. The aim of the present study was to investigate the mRNA expression of 5 alpha-reductase 1 as well as 3 alpha-HSD 1, 2, 3 and 20 alpha-HSD in brain tissue from patients with pharmacoresistant temporal lobe epilepsy (TLE). Specimens were derived from either the hippocampus or the temporal lobe cortex and from the tumor-free approach corridor tissue of patients with brain tumors. Quantification of different mRNAs was achieved by real time PCR. In addition, we provide data on simultaneous evaluation of serum AP concentrations. We could demonstrate that 3 alpha-HSD 1 was not expressed in the hippocampus and temporal lobe of patients with TLE. In the hippocampus and temporal lobe, the expression levels of 3 alpha-HSD 2 were about 20% of that in liver tissue, those of 3 alpha-HSD 3 about 7% and those of 20 alpha-HSD about 2%, respectively. In patients with TLE, expression of 3 alpha-HSD 2 was significantly higher in the hippocampus than in temporal lobe cortex tissue (P<0.006). AP concentrations did not correlate significantly with the mRNA expression levels of 5 alpha-reductase 1, 3 alpha-HSD 2 and 3 and 20 alpha-HSD in any of the patient groups under investigation. In conclusion, the present study demonstrates mRNA expression of 5 alpha-reductase 1 and 3 alpha-HSD 2 and 3 and 20 alpha-HSD in the hippocampus and temporal lobe of epileptic patients. These findings provide further molecular biological evidence for the formation and metabolism of neuroactive steroids in the human brain. PMID- 12742592 TI - Neonatal convulsions and epileptic encephalopathy in an Italian family with a missense mutation in the fifth transmembrane region of KCNQ2. AB - Mutations in the voltage gated K(+)-channel gene KCNQ2 are known to cause benign familial neonatal convulsions (BFNC), which are characterized by a benign course, spontaneous remission and normal psychomotor development. Most KCNQ2 mutations can be predicted to truncate the protein. Only a few amino acid exchanges have been found, and their localization was restricted to either the pore region or the fourth or sixth transmembrane region (TM). We have now identified the first KCNQ2 mutation located within TM5, affecting a highly conserved serine in amino acid position 247 of the predicted protein. The clinical history of the two affected family members is not compatible with typical BFNC. The poor outcome in the index patient raises the question if at least some KCNQ2 mutations might increase the risk to develop therapy-resistant epilepsy. Additional studies are needed to evaluate the possibility of a causal relationship between KCNQ2 mutations and severe early infantile epilepsy. PMID- 12742593 TI - Inter-ictal and post-ictal circulating levels of allopregnanolone, an anticonvulsant metabolite of progesterone, in epileptic children. AB - Allopregnanolone belongs to a group of neuroactive steroid hormones, or neurosteroids, synthesized and acting within the brain and is as a potent endogenous positive modulator of GABA(A) receptor complex. Administration of allopregnanolone protects rats against pentylentetrazol, bicuculline, kainic acid, and picrotoxin-induced seizures. We investigated serum allopregnanolone levels in children with active epilepsy at pubertal Tanner's stage I (n=52). Blood specimens were collected at least 12 h after a seizure (inter-ictal). In a subgroup of patients (n=11), specimens were also collected within 30 min from a seizure attack (post-ictal). Healthy age-matched children (n=18) served as controls. Serum allopregnanolone was measured by radioimmunoassay using a polyclonal antiserum. The inter-ictal serum allopregnanolone levels in the epileptic children were not statistically different from those detected in the control group, whereas post-ictal levels were significantly higher than the inter ictal ones (P=0.0001). In this subgroup of patients allopregnanolone levels decreased to the basal values during the following 12 h. Serum allopregnanolone levels may therefore reflect changes in neuronal excitability, and allopregnanolone appears to be a reliable circulating marker of epileptic seizures. It is possible that increased post-ictal serum levels of allopregnanolone may play a role in modulating neuronal excitability and may represent an endogenous mechanism of seizure control. PMID- 12742594 TI - Rubella immunoglobulin G antibody titers in children with seizures. AB - The third National Health and Nutrition Examination Survey was utilized to assess serum rubella immunoglobulin G antibody titers in a representative sample of American children. Antibody titers were significantly lower in children with a history of seizures and in children treated for seizures compared to unaffected children, after adjustment for age, ethnicity, residence, and region (P=0.022 and 0.029, respectively). Children with the lowest antibody titers were non-Hispanic whites who had a history of seizures. The percentage of US adolescents and young adults with a history of seizures that may have insufficient immunity to rubella is estimated to be 14-50%. PMID- 12742595 TI - Calorie restriction of a high-carbohydrate diet elevates the threshold of PTZ induced seizures to values equal to those seen with a ketogenic diet. AB - The purpose of this study was to evaluate the contributions of ketonemia, caloric restriction, and carbohydrates to seizure protection in rats fed selected diets. Male Sprague-Dawley rats were fed experimental diets of two basic types, one high in carbohydrates and restricted to 90, 65, or 50% of the normal daily caloric requirement and the other a normal rodent chow diet restricted to 90 or 65% of the daily caloric requirement. After consuming their respective diets for 20 days, animals were subjected to tail-vein infusion of pentylenetetrazole (PTZ) to determine seizure threshold, taken as the dose required to evoke the first clonic reaction. Seizure thresholds were compared to those of rats fed control diets of either normal rodent chow fed ad libitum or a standard high-fat (ketogenic) diet calorie-restricted to 90% of daily caloric requirement, all animals age- and weight-matched at the time of diet onset. All diets were balanced for vitamins and minerals and contained at least 10% protein (by weight). Seizure threshold and ketonemia were elevated in both experimental diets in approximate proportion to the degree of calorie restriction. Animals fed the most severely restricted high-carbohydrate diet (50%) had seizure thresholds equal to those fed the ketogenic diet but had significantly lower ketonemia. PMID- 12742596 TI - The lack of association between febrile convulsions and polymorphisms in SCN1A. AB - Febrile convulsions (FCs) represent the majority of childhood seizures, and patients have a genetic predisposition to their development. The genetic susceptibility to FCs seems to involve multiple genes in most instances. Recent studies provided evidence that mutations in SCN1A represent the most frequent cause of generalized epilepsy with febrile seizures plus an autosomal-dominant epilepsy syndrome. SCN1A mutations alter channel inactivation, resulting in persistent inward sodium current. It is not known if polymorphisms in those genes involved in familial epilepsies also contribute to the pathogenesis of FCs. By performing an association study, we used single nucleotide polymorphisms to investigate the distribution of genotypes of SCN1A in patients with FCs. A total of 104 Taiwanese children with FCs and 83 normal control subjects were included in the study. Polymerase chain reaction was used to identify the A/G polymorphism of the SCN1A gene. The results showed that genotypes and allelic frequencies for the SCN1A gene polymorphisms in both groups were not significantly different. These data suggest that the SCN1A gene might not be one of the susceptibility factors for FCs. Pure FCs and febrile convulsions associated with idiopathic generalized epilepsy may not share a common genetic etiology. PMID- 12742597 TI - 3-D reconstructed magnetic resonance imaging in localization of subdural EEG electrodes. Case illustration. AB - We report an illustrative case of presurgical evaluation for epilepsy surgery, where the three-dimensional reconstructed magnetic resonance imaging played a pivotal role in determining the exact location of the subdural strip electrodes in temporomesial area. The tip of one the frontal strip electrodes was actually recording the temporopolar ictal activity. This contributed conclusively to the decision for surgical treatment and to the excellent outcome. PMID- 12742598 TI - Topiramate protects against glutamate- and kainate-induced neurotoxicity in primary neuronal-astroglial cultures. AB - Potential neuroprotective effects of the antiepileptic drug (AED) topiramate (TPM) were evaluated using primary neuronal-astroglial cultures or astroglial enriched cultures from newborn rats exposed to excitotoxic concentrations of glutamate (Glu) or kainate. Neurons expressed functional Glu receptors of the NMDA and AMPA/kainate types as evaluated by immunocytochemistry and Ca(2+) imaging. When Glu (10 mM) was added to 9-10-day cultures incubated with the fluorescent dye calcein/AM for 5h, there was a marked cell loss in both culture types, but was more pronounced in the neuronal-astroglial cultures. When TPM (5 10 microM) was included in the medium together with Glu, the amount of surviving cells was significantly higher in the neuronal-astroglial cultures, but not in the astroglial-enriched cultures. Immuno-labeling of the cultures revealed an enhanced survival of MAP positive neuronal cells when TPM was included in the Glu containing medium. As TPM has a proven negative modulatory effect on kainate activated receptors, neuronal-astroglial cultures were further exposed to excitotoxic concentrations of kainate (100 microM) and analyzed immunohistochemically. Significantly more MAP positive neurons survived in the TPM containing medium and showed a morphology similar to untreated cells. Valproate and phenytoin were used as reference AEDs. In conclusion, our results demonstrate a protective effect of TPM upon neuronal cells in primary culture, exposed to excitotoxic levels of Glu or kainate. PMID- 12742601 TI - Social capital, life expectancy and mortality: a cross-national examination. AB - This paper analyses the relationship between social capital and population health. The analysis is carried out within an econometric model of population health in 19 countries in the Organisation for Economic Co-operation and Development countries using panel data covering three different time periods. Social capital is measured by the proportion of people who say that that they generally trust other people and by membership in voluntary associations. The model performs well in explaining health outcomes. We find very little statistically significant evidence that the standard indicators of social capital have a positive effect on population health. By contrast, per capita income and the proportion of health expenditure financed by the government are both significantly and positively associated with better health outcomes. The paper casts doubt upon the widely accepted hypothesis that social capital has a positive effect on health and illustrates the importance of testing this kind of hypothesis in an extended model. PMID- 12742600 TI - Normalizing the exceptional: incorporating the "abortion pill" into mainstream medicine. AB - Mifepristone, also known as RU-486, and in the US known as "the French abortion pill", finally received FDA approval in the United States in September 2000. This paper discusses the steps now in process to integrate this drug into mainstream healthcare and the sociological implications of those efforts. Each of the steps that is normally taken to introduce a newly approved medication in the US context is rendered highly complex in the case of mifepristone--because of the unique circumstances of abortion in both American culture generally, and medical culture specifically. The story of RU-486/mifepristone, as it is currently unfolding, can be understood as one of attempting to "normalize the exceptional". After offering a brief historical overview of the protracted struggle for FDA approval of mifepristone in the US, this paper discusses the typical processes for integration of a newly approved medication into mainstream medicine and contrasts this process with the special challenges posed by a drug that is associated with abortion. We outline the challenges to implementation, including both external and internal obstacles. We compare the traditional role of a pharmaceutical company in drug diffusion and the circumstances of the company that produces mifepristone in the US. We discuss such external obstacles as the conflict between the FDA-approved regime and an evidence-based alternative; the necessity for physicians to order and dispense this drug; the ambiguity over the need for ultrasonography; and insurance reimbursement, malpractice, and other legal issues. Internal issues addressed include "turf issues" between medical specialties and between physicians and advanced practice clinicians as well as concerns over "cowboy medicine", and patient compliance. This paper concludes with an exploration of the sociological implications of this effort to "normalize the exceptional". PMID- 12742599 TI - Elevated extracellular levels of glutamate, aspartate and gamma-aminobutyric acid within the intraoperative, spontaneously epileptiform human hippocampus. AB - We report preliminary results from four patients subjected to hippocampal electrocorticography and microdialysis during temporal lobe epilepsy surgery. In two cases, spontaneously vigorous hippocampal epileptiform activity (EA) was identified; basal dialysate levels for hippocampal glutamate, aspartate, and gamma-aminobutyric acid ranged from approximately 23- to 84-fold, 19- to 33-fold and 10- to 34-fold higher, respectively, compared to the two cases of minimal hippocampal EA. These findings represent the first intraoperative evidence of elevated extracellular levels of neuroactive amino acids within the spontaneously epileptiform human hippocampus. PMID- 12742602 TI - Investments in social capital--implications of social interactions for the production of health. AB - This paper develops a theoretical model of the family as producer of health- and social capital. There are both direct and indirect returns on the production and accumulation of health- and social capital. Direct returns (the consumption motives) result since health and social capital both enhance individual welfare per se. Indirect returns (the investment motives) result since health capital increases the amount of productive time, and social capital improves the efficiency of the production technology used for producing health capital. The main prediction of the theoretical model is that the amount of social capital is positively related to the level of health; individuals with high levels of social capital are healthier than individuals with lower levels of social capital, ceteris paribus. An empirical model is estimated, using a set of individual panel data from three different time periods in Sweden. We find that social capital is positively related to the level of health capital, which supports the theoretical model. Further, we find that the level of social capital (1) declines with age, (2) is lower for those married or cohabiting, and (3) is lower for men than for women. PMID- 12742603 TI - The methodology of HIV/AIDS impact studies: a review of current practices. AB - This review of studies on the socio-economic impact of HIV/AIDS shows that diversity in methodological design, which often is a result of practical considerations and resource constraints rather than of poor design, is the norm. This limits the comparability of research findings. More detailed reporting on method, which is not the norm, can go some way towards facilitating such comparison. Furthermore, the review underlines the importance of exploring intervention issues in more detail. Researchers need to employ results in answering specific policy questions. Scope remains for more impact studies to be conducted in developing countries in general and in certain high prevalence countries in specific, i.e. Southern Africa. Studies that explore the urban/rural dynamics of and clients' perceptions and behavior in seeking care and support are necessary to better understand the epidemic. The role of community-based organizations, non-governmental organizations and other stakeholders in studies of this nature can be expanded. Larger studies generally have more statistical power, but smaller, in-depth studies can be equally valuable. A careful stratification of sample populations can enhance the quality of cross-sectional studies. Qualitative methods should be used to complement the current reliance on survey-based methods of data collection. More longitudinal studies are required to explore the long-term impacts of HIV/AIDS. HIV/AIDS training for fieldworkers should be standard in studies of this nature, while cognizance should be taken of the dangers of employing local people as fieldworkers in studies of such sensitive nature. Scope remains for the further empirical analysis of data from impact studies, which requires these data sets being made accessible to more researchers. In the longer term, an attempt at standardizing core modules in impact studies can help to improve our understanding of the impact of HIV/AIDS in different settings. PMID- 12742604 TI - The rule of rescue. AB - Jonsen coined the term "Rule of Rescue"(RR) to describe the imperative people feel to rescue identifiable individuals facing avoidable death. In this paper we attempt to draw a more detailed picture of the RR, identifying its conflict with cost-effectiveness analysis, the preference it entails for identifiable over statistical lives, the shock-horror response it elicits, the preference it entails for lifesaving over non-lifesaving measures, its extension to non-life threatening conditions, and whether it is motivated by duty or sympathy. We also consider the measurement problems it raises, and argue that quantifying the RR would probably require a two-stage procedure. In the first stage the size of the individual utility gain from a health intervention would be assessed using a technique such as the Standard Gamble or the Time Trade-Off, and in the second the social benefits arising from the RR would be quantified employing the Person Trade-Off. We also consider the normative status of the RR. We argue that it can be defended from a utilitarian point of view, on the ground that rescues increase well-being by reinforcing people's belief that they live in a community that places great value upon life. However, utilitarianism has long been criticised for failing to take sufficient account of fairness, and the case is no different here: fairness requires that we do not discriminate between individuals on morally irrelevant grounds, whereas being "identifiable" does not seem to be a morally relevant ground for discrimination. PMID- 12742605 TI - From damaged nerves to masked depression: inevitability and hope in Latvian psychiatric narratives. AB - Psychiatric language in Latvia has been invaded by the diagnosis of depression and masked depression. Depression has been promoted by the translation into Latvian of the International Classification of Diseases and by conferences organized by pharmaceutical companies and aimed at educating psychiatrists and family doctors about the new diagnostic categories. The language of depression represents a radical departure from older languages of somatic distress that were central both to Soviet Psychiatry and to lay conceptualizations of distress. However, the new practitioners who favour the diagnosis of depression have a highly atomistic and culture blind approach to patients' problems. In order to selectively cleanse the presentation of distress, various strategies for eliminating social context and suppressing patients' narratives are used during psychiatric consultations. Alongside these imported psychiatric languages, recognition of the physically and socially embedded nature of human experience and its historicity persist. Not all psychiatrists eliminate subjective narrative from the consultation dialogue. However, prioritizing mental over physical states is not linked in a straightforward way to other dualisms such as the intentional versus the accidental and the voluntary versus the involuntary. For many depressed patients autonomy is restricted to being a good patient and learning about their condition. Conversely, psychiatrists who start out by addressing their patients' physical discomfort may move on to open up a range of narrative possibilities. PMID- 12742606 TI - Using burden of disease information for health planning in developing countries: the experience from Uganda. AB - Given the growing interest in both the use of evidence in planning and in using the burden of disease measure (BOD) and cost-effectiveness analysis, we explored health planners' perception of the usefulness of the BOD in priority setting and planning in developing countries, using Uganda as an example. An exploratory qualitative approach involving in-depth interviews with key policy makers in health at district and national levels was employed. Interviews were supplemented with a review of relevant documents. Analysis involved identification of key concepts from the interviews. Concepts were grouped into categories, namely, the appeal of quantitative data, data limitations, opaque methodology, planning as a political process and opportunity costs. These form the basis of this article. We found that the BOD study results have been used as the basis for the national health policy and in defining the contents of the national essential health care package. The quantification and ranking of disease burden is appreciated by politicians and used for advocacy, resource mobilization and re-allocation. The results have also provided information for priority setting and strategic planning. Limitations to its use included poor understanding of the methodology, poor quality of data in-puts, low involvement of stakeholders, inability of the methodology to capture key non-economic issues, and the costs of carrying out the study. There is commitment, by Ugandan planners to using evidence in priority setting. Since this was an exploratory study, there is a need for more studies in developing countries to document their experiences with the use of evidence, and specifically, the BOD approach in planning and priority setting. Such information would contribute to further synthesis of the approach. PMID- 12742607 TI - Does low reading achievement at school entry cause conduct problems? AB - Conduct problems place children at increased risk for a broad array of negative health and social outcomes that include conduct disorder, injuries and violence, school failure, substance abuse, depression, and suicide. Prevention interventions have the potential to interrupt the chain of events linking early conduct problem symptoms to future negative life outcomes, but have received much less emphasis than interventions designed to treat established cases of disorder. Reading problems are a well-established correlate of conduct disorder. However, whether or not reading problems cause conduct disorder continues to be debated. If they are in fact a causal risk factor this would justify the design and evaluation of interventions designed to enhance reading skills and/or remediate problems. In this paper we use logistic regression techniques to evaluate the relation between reading achievement at school entry and conduct problems 30 months later, in a representative, non-clinic sample of kindergarten and grade one children, in Ontario, Canada. The findings show that an eight point increase in reading scores (equivalent to an moderate effect size of 0.5) would result in a 23 per cent decrease in the risk of conduct problems 30 months later, after controlling for gender, income and baseline conduct problem symptoms. We conclude that reading problems may contribute to the early onset of conduct disorder. Randomized experimental studies designed to evaluate the effects of reading programmes in non-clinic samples of children are needed to: (i) establish whether the link between reading problems at school entry and conduct disorder is causal; and (ii) determine whether reading intervention programmes are an effective conduct disorder prevention strategy. PMID- 12742608 TI - Pregnant smokers who quit, pregnant smokers who don't: does history of problem behavior make a difference? AB - More than half of women who smoke in the USA continue to do so while pregnant. While socioeconomic and demographic factors that distinguish pregnancy quitters from persistent smokers have been identified, less is known about behavioral factors that are associated with persistent smoking. Because smoking during pregnancy is not only an individual, but also a maternal behavior, it may have different behavioral correlates than women's smoking has in general. We propose a conceptual framework in which smoking during pregnancy is viewed as a maternal problem behavior. We explore this conceptualization by examining whether persistent smoking during pregnancy is associated with a pattern of psychosocial risk- and health-compromising behaviors in multiple domains, with pilot data from a small clinic-based sample. Data are presented for 96 predominantly Caucasian, working-class pregnant women recruited from prenatal clinics in the USA. Smoking during pregnancy was measured repeatedly by self-report and biochemical assay. Participants were non-smokers (37%), pregnancy quitters (17%), and persistent smokers (46%). These groups were compared in terms of their history of problem behavior in three domains: interpersonal difficulties, problems in adaptive functioning and problematic health behaviors. With few exceptions, smokers were more likely to have problematic relationships, poorer adaptive functioning and to engage in problematic health behaviors, than both pregnancy quitters and non smokers. This pattern of problem behavior may interfere with the effectiveness of standard public health prenatal cessation interventions for a sub-group of women. Examining pregnancy smoking as part of a broader matrix of problem behavior may help to identify pregnant women most at risk for persistent smoking and inform the development of targeted interventions. PMID- 12742609 TI - Evaluation of primary health care reform in Estonia. AB - Estonia began to reform its health care system by reorganizing primary health care (PHC). Ten years ago, the health care system was inefficient, supply was biased towards institutional care, and PHC was almost non-existent in the western understanding: it was provided by different specialists in policlinics, and the financial basis of the system was in crisis. The reform had the following aims: to develop PHC by introducing family medicine, to guarantee the whole population access to family doctors' services, and to secure stable funding for these services. In 1998, a new phase in the reform was introduced through the creation of a new funding system for primary care services. The aim of this paper is to present a practically applicable set of indicators to evaluate PHC reform in terms of health economics criteria and then to apply these indicators in evaluation of the Estonian PHC reform. PMID- 12742610 TI - Community stress, demoralization, and body mass index: evidence for social signal transduction. AB - Quantification of the relationship between community-level chronic stress from neighborhood conditions and individual morale has rarely been reported. In this work, pregnant women were recruited at the prenatal clinics of Harlem Hospital and Columbia Presbyterian Medical Center in the USA, and given an initial questionnaire that included all 27 questions of the Dohrenwend demoralization instrument, as well as questions about household economics and health. An index of chronic community stress (ICCS) was compiled for each of the health areas of the study zone by standardizing and weighting each stressor significantly associated with low birthweight rate and summing the standardized, weighted values. Health areas were divided into ICCS quintiles. The graph of the quintile weighted averages of the index vs. the quintile averages of the demoralization score was an asymmetric inverted 'U' shape that fitted well to a stochastic resonance signal transduction model (adjusted R2=0.73). On average, the women in the worst three quintiles were much heavier than those of the two best quintiles. Women reporting household economic deprivations were significantly more demoralized than the others. Median health area rents were strongly negatively associated with the ICCS. The worst average demoralization score occurred in the middle quintile, a state of coping with both poor community conditions and an economically strained household. Rents bridge community conditions and household economics. PMID- 12742611 TI - Variation in physicians' definitions of the competent parent and other barriers to guideline adherence: the case of pediatric minor head injury management. AB - A lack of consensus regarding the definition of even an everyday term can affect physician adherence to clinical guidelines using that term. We demonstrate this by taking, as an illustrative case, the American Academy of Pediatrics' minor head injury (MHI) management guidelines, which generally recommend at-home observation by a "competent" parent (or the equivalent). The recommendation assumes consensus among physicians as to what parental competence comprises. We systematically examined this assumption. Physicians associated with Children's Hospital, San Diego were mailed a survey asking them to freely list terms defining parental competence. Independent variables were: physician gender, training, specialty, practice location, patient age mix, years in practice, and number of MHI cases seen per year. Dependent variables were: free-list content and length, ease and style of competence decision-making style (e.g., independent or collaborative), familiarity with the guidelines, and likelihood of ordering a computed tomography (CT) scan. Of 112 respondents, 34 (30%) were "not at all" or only "slightly" familiar with the guidelines; 23% (21/112) "frequently" or "sometimes" ordered CTs when the guidelines did not support this. Surgeons were more likely to order discretionary CTs. Office-based, pediatric-trained, and female physicians each found it significantly easier than their counterparts to determine which parents were, in their opinion, competent. Men reported using "objective" criteria significantly less frequently than women. A total of 64 discrete criteria were listed. Individual lists contained an average of 6.5 items. Surgeon's lists were significantly shorter than those of medical physicians. Seven sub-domains of competence were identified. Parental competence is not a unitary construct interpreted similarly by all physicians. Subgroups with distinct models may exist; training and specialization may be key variables. To decrease MHI management variation, guidelines should specify parental competence factors to be considered; they may need to be tailored to different physician subgroups. PMID- 12742612 TI - Country of birth, instrumental activities of daily living, self-rated health and mortality: a Swedish population-based survey of people aged 55-74. AB - There is scant knowledge of the effects of country of birth on the health of individuals in the years prior to and after retirement. The aim of this study was to consider country of birth in relation to health status, instrumental activities of daily living (IADL) and all-cause mortality when adjusted for socioeconomic status (SES). Cross-sectional data were collected between 1986 and 1991 on 8959 individuals between the ages of 55 and 74. Self-reported data were analysed using a logistic regression model while the mortality data were analysed by means of a proportional hazard model. In the present study, immigrants from Southern Europe, Eastern Europe and Finland carried significantly increased risks of poor health even after adjustment for SES. Southern Europeans, refugees from Developing countries and Finns exhibited an increased risk of impaired IADL compared to Swedes, even after adjustment for SES. In conclusion, country of birth was associated with poor health status and impaired IADL. This association remained after adjustment for SES. In accordance with pre-study expectations, mortality was predicted by impaired IADL and male gender. Country of birth was not associated with all-cause mortality. PMID- 12742613 TI - Sociodemographics, self-rated health, and mortality in the US. AB - Using data from the 1987 National Medical Expenditure Survey, a representative sample of US civilians, and their 5-year mortality, we examined the adjusted relationships among baseline self-reported health, derived from SF-20 subscales (health perceptions, physical function, role function and mental health) and sociodemographics (age, sex, race/ethnicity, income and education) and subsequent mortality. Included were 21,363 persons aged 21 and over, with complete follow-up on 19,812. Physical function showed the greatest decline with age, whereas mental health increased slightly. Women reported lower health for all scales except role function. Greater income was associated with better health, least marked for mental health. Greater education was associated with better health, most marked for health perceptions. Compared with whites, blacks reported lower health, whereas Latinos reported higher health. Lower self-reported health predicted increased adjusted mortality. After adjustment for baseline self-rated health, the relationships between income and education and mortality were greatly attenuated, whereas the relationships between age, gender, race/ethnicity and mortality were not. Self-rated health exhibited more profound relationships with mortality in younger persons, those with more education, and whites. In conclusion, lower socioeconomic status (SES), and being black are associated with lower reported health status and higher mortality; women report lower health status but exhibit lower mortality; and Latinos report higher health status and exhibit lower mortality. The effects of SES on mortality are largely explained by their associations with self-rated health, whereas, the effects of gender and race/ethnicity on mortality appear to act through independent pathways. Because of these differential sociodemographic relationships caution is urged when using self-rated health measures in research, clinical, and policy settings. PMID- 12742614 TI - Civil war and child health: regional and ethnic dimensions of child immunization and malnutrition in Angola. AB - This study arises from a general proposition that different levels and types of exposure to war are crucial in shaping health outcomes in a population under war induced duress. We analyze civil war-related regional and ethnolinguistic differentials in age-adequate immunization (complete vaccination for age) and levels of malnutrition in Angola. Our analysis is based on data from a nationally representative survey conducted in 1996, some 2 years after the end of one of the most destructive periods of hostilities in the history of Angolan civil war. The data show that despite Angola's unique mineral wealth, the nation's levels of child age-adequate immunization is lower and malnutrition rates are higher than in most of sub-Saharan Africa. To examine age-adequate immunization and chronic malnutrition we fit logistic regression models that include the regional degree of war impact and ethnolinguistic group, in addition to rural-urban differences and other conventional sociodemographic characteristics. The tests reveal a significant disadvantage of rural children relative to urban children in both immunization and chronic malnutrition. Net of the rural-urban differences, we also detect a significant disadvantage of children residing in parts of the country that had been most affected by the fighting. The tests also point to a lower level of immunization and higher level of chronic malnutrition among children from the ethnolinguistic group commonly identified with the opposition. These associations tend to be stronger among children who were born and/or grew up during war than among children who were born after peace was re-established. PMID- 12742615 TI - Politics of leprosy segregation in Japan: the emergence, transformation and abolition of the patient segregation policy. AB - The segregation of leprosy patients, a practice introduced early in the 20th century, was maintained in Japan after World War II. It locked in the viability of subsequent policy choices, and patients' isolation was sustained long after it was proven to be scientifically unnecessary. For leprologists and leprosarium directors, there was little opportunity to conceptualize and test the epidemiological validity and effectiveness of outpatient services as alternatives to the existing policy, since most of the patients were already hospitalized. Since leprosy was no longer a threat to the general public, bureaucratic officials, as well as legislators, lacked strong incentives to reformulate the overall policy. Within the Ministry of Health and Welfare, daily tasks were largely transferred to the section for leprosarium management, and the search for other options lost importance. For patients, long institutionalization elevated their dependency on life in leprosaria. These conditions must be emphasized as policy legacies, the results of past policies, since they posed obstacles to effective policy innovation in accordance with changing scientific knowledge. To make policies reflective of scientific knowledge, it is essential to understand and foresee the effect of policy legacy, when introducing and appraising public health policies. PMID- 12742616 TI - A spatial analysis of county-level variation in hospitalization rates for low back problems in North Carolina. AB - Hospitalization rates for low back problems vary widely. In previous non-spatial analyses, population-level socioeconomic and health resource characteristics have explained little of the variation in rates. This study examines geographic variation in hospitalization rates for low back problems while controlling for spatial dependence in the data. County-level surgical and medical hospitalization rates were calculated using North Carolina hospital (USA) discharge data from 1990-92. Non-spatial and spatial regression models were estimated using socioeconomic and health resource predictors. Both surgical and medical rates varied significantly among the 100 counties. Non-spatial models explained 62% of variation in log-transformed surgical rates and 66% of variation in log transformed medical rates; however, residuals showed significant spatial dependence. Spatial lag models were therefore applied. Using simple contiguity spatial weights, surgery rates increased with higher percent urban population, primary care physician density, and discharge rate for other causes, and decreased with higher percent college graduates, percent disabled, occupied hospital bed density, and unoccupied hospital bed density. There was a nonlinear relationship between surgery rates and percent employed in heavy lifting/transportation industries. Medical rates increased with higher other cause discharge rate and with MRI/CT scanner availability, and decreased with higher percent urban population, percent nonwhite population, percent in heavy lifting/transportation industries, and unoccupied hospital bed density. The results show that population-level socioeconomic and health resource characteristics are important determinants of variation in low back hospitalization rates. Independent of these variables, a separate spatial process produces geographic clustering of high-rate counties. Spatial effects are important and should be considered in small area analyses. PMID- 12742617 TI - Understanding long-standing illness among older people. AB - This paper describes a study of older people's interpretations of a survey question about long-standing illness, disability or infirmity. This and similar questions are frequently used in various studies and surveys that influence policy and planning. With population ageing and growing concern about the health of older people, we sought to examine the survey question's relevance to older people. Following-on from a cross-sectional survey of 999 people aged 65 and over in the UK, we explored their interpretations of the survey item by asking it in the context of in-depth interviews with 24 respondents. We found that few of our respondents subscribed to the constructs of long-standing illness, disability or infirmity that surveys often employ. Older people's descriptions of their health status in response to a "standard-issue" survey question are informed by their understandings of health itself as well as elements such as control, engagement with health service providers, time and ageing. This implies that questionnaire based surveys may not only be unable to capture the meaning of chronic illness to older people, but also its prevalence. We conclude on a methodological note by reminding ourselves that answers to survey and in-depth interviews are narratives told about health status. As such, both represent many facets of social and cultural life, and are best assessed as "trustworthy" rather than "truthful". PMID- 12742618 TI - Response to Gilbert & Walker 'treading the path of least resistance'. PMID- 12742619 TI - The influence of chemosensory input and gonadotropin releasing hormone on mating behavior circuits in male hamsters. AB - Chemosensory input is important for mating behavior in male hamsters. Chemosignals found in female hamster vaginal fluid activate regions of the brain that receive input from the vomeronasal/accessory olfactory system and are important for mating behavior. Mating or exposure to these chemosignals produces increased Fos protein expression in the amygdala, bed nucleus of the stria terminalis, and medial preoptic area (MPOA). These brain regions contain cell bodies and/or fibers of gonadotropin releasing hormone (GnRH) neurons, suggesting potential relationships between chemosensory systems and GnRH. GnRH is released naturally when male rodents (mice and hamsters) encounter female chemosignals, and intracerebrally injected GnRH restores mating behavior in sexually naive male hamsters after removal of the vomeronasal organs. We report here that the combination of pheromone exposure and intracerebrally-injected GnRH increases Fos expression in the MPOA above the increase seen in pheromone-exposed males, or in males given only the exogenous GnRH. In males with vomeronasal organs removed (VNX), there was an also an increment in Fos expression in the MPOA when these pheromone exposed males were injected with GnRH, provided they had previous sexual experience. Males with vomeronasal organs removed and without sexual experience showed increased Fos expression in the medial amygdala when pheromone exposure and GnRH injection were combined, but not in the medial preoptic area. PMID- 12742620 TI - Transplanted neuronal progenitor cells in a peripheral nerve gap promote nerve repair. AB - A basic experiment of peripheral nerve regeneration using neuronal progenitor cells embedded in collagen gel was performed in a rat sciatic nerve defect. First, when neuronal progenitor cells derived from the fetal rat hippocampus were cultured in atelocollagen-containing medium, neurospheres positive for anti nestin antibody were confirmed after 8 days. These cells differentiated into astrocytes positive for anti-glial fibrillary acidic protein (GFAP) antibody, oligodendrocytes positive for anti-galactocerebroside (GalC) antibody and neurons positive for anti-neurofilament 200 (NF200) antibody, and they were capable of extending axons. They also differentiated into Schwann-like supportive cells positive for anti-s100 and anti-p75 antibody. Next, a 15-mm defect was prepared in the sciatic nerve of mature rats, and the nerve was bridged with a silicone tube filled with neuronal progenitor cells (1 x 10(5)) embedded in collagen gel. The transplanted neuronal progenitor cells were labeled in advance with 5-bromo-2 deoxyuridine (BrdU). When the regenerated tissue was examined 6 weeks and 10 weeks after grafting, the number and diameter of myelinated fibers were significantly increased compared with a control tube without neuronal progenitor cells. Action potentials were detected in the regenerated nerve. Also, cells positive for both anti-BrdU antibody and anti-S100 or anti-p75 antibody were observed in the regenerated tissue, and part of the grafted neural stem cells were considered to have differentiated into Schwann cell-like supportive cells. From these results neuronal progenitor cells derived from the fetal rat hippocampus are considered to retain their proliferative and differentiating abilities in collagen gel, and when transplanted to a site of peripheral nerve defect, part of them differentiate into supportive cells and they contributed to promotion of axonal regeneration. PMID- 12742621 TI - Immunocytochemical localization of the alpha3, alpha4, alpha5, alpha7, beta2, beta3 and beta4 nicotinic acetylcholine receptor subunits in the locus coeruleus of the rat. AB - The presence of nicotinic acetylcholine receptors (nAChRs) within the locus coeruleus (LC) has been examined using a wide range of techniques. However, the expression pattern of individual nicotinic receptor subunits has not been described. Using immunocytochemistry, we demonstrate the distribution of the alpha3, alpha4, alpha5, alpha7, beta2, beta3 and beta4 nAChR subunits within the LC. Most nAChR subunits were expressed on neuronal perikarya within the LC nucleus. The alpha3, alpha4, alpha7 and beta3 immunoreactive neurons were evenly distributed in the dorsal and ventral LC whereas the alpha5, beta2 and beta4 nAChR subunits were preferentially confined to the upper dorsal section. In addition to neuronal perikarya, alpha4, alpha5 and beta2 immunoreactive fibers were observed. With the exception of the alpha3 subunit, punctate labeling was observed within and immediately surrounding the LC. These data are consistent with the presence of multiple nAChRs within the LC and extend these findings to show the distribution pattern of each nAChR subunit throughout the LC nucleus. PMID- 12742622 TI - Kinetics of creatine in blood and brain after intraperitoneal injection in the rat. AB - Creatine has in recent years raised the interest of the neurologist, because it has been used in children with hereditary disorders of creatine metabolism and because experimental data suggest that it may exert a protective effect against various neurological diseases including stroke. Moreover, it is widely used as a nutritional supplement. It is well known that creatine crosses the blood-brain barrier with difficulty, however its accumulation into the brain after systemic administration is still not completely known. In the present experiments we studied its accumulation into rat brain tissue after intraperitoneal (i.p.) single or repeated injections. After a single injection of 160 mg/kg, radioactively labelled creatine (14C-creatine) entered the brain to a limited extent. It reached a plateau value of around 70 microM above baseline, that remained stable for at least 9 h. This amount of exogenous creatine obviously added to the endogenous creatine store. This increase is a minor one, since endogenous creatine has a brain concentration of about 10 mM. In accordance with this conclusion, when single or repeated injections of unlabelled ('cold') creatine were administered to rats, no sizable increase could be measured with high-performance liquid chromatography in the brain levels of either this compound or its phosphorylated derivative, phosphocreatine. Although our data clearly show some passage of serum creatine into the brain, other strategies are needed to improve passage of creatine across the blood-brain barrier in a way that it may be suitable to treat acute conditions like stroke. PMID- 12742623 TI - Efferent subcortical projections of the laryngeal motorcortex in the rhesus monkey. AB - In order to better understand the descending voluntary vocal control pathway, the efferent subcortical projections of the laryngeal motorcortex were studied in the rhesus monkey (Macaca mulatta). For this purpose, the left motorcortex was exposed in three animals under narcosis. By electrical brain stimulation, sites were identified yielding vocal fold adduction. Effective sites were injected with the anterograde tracer biotin dextran amine. Subcortical projections could be traced within the forebrain to the putamen, caudate nucleus, claustrum, zona incerta, field H of Forel and a number of thalamic nuclei, with the heaviest projections to the nuclei ventralis lateralis, ventralis posteromedialis, including its parvocellular part, medialis dorsalis, centralis medialis, centrum medianum and reuniens. In the midbrain, labeling was found in the deep mesencephalic nucleus. In the lower brainstem, fibers terminated in the pontine and medullary reticular formation, locus coeruleus, nucleus subcoeruleus, medial parabrachial nucleus, nucleus of the spinal trigeminal tract, solitary tract nucleus and facial nucleus. No projections were found to the nucl. ambiguus. The fact that monkeys, in contrast to humans, lack a direct connection of the motorcortex with the laryngeal motoneurons suggests that this connection has evolved in the last few million years and might represent one of the factors that made speech evolution possible. PMID- 12742624 TI - Surface and intracellular nicotinic receptors expressed in intact adrenal chromaffin cells: direct measurements using [3H]epibatidine. AB - The presence and importance of assembled, intracellular neuronal nicotinic acetylcholine receptors (nAChRs) has not been established in native systems. In these studies [3H]epibatidine binding techniques were used to characterize surface and intracellular sites expressed in intact bovine adrenal chromaffin cells in culture. Permeant (300 microM nicotine) and impermeant (5 mM carbachol) cholinergic agents were used to define specific [3H]epibatidine binding to total (surface and intracellular) sites and surface sites, respectively. Intracellular [3H]epibatidine binding sites were characterized after eliminating surface binding sites via alkylation. Equilibrium binding to all sites was reached within 30 min at room temperature. Homologous (epibatidine) competition experiments on total (surface and intracellular) binding sites demonstrated a significant fraction of the high affinity sites were localized to intracellular compartments. Saturation binding assays to surface and intracellular sites revealed K(d) values of 1.9+/-1.1 and 3.6+/-1.9 nM, respectively. These binding studies document the existence of a significant population of high affinity, intracellular binding sites in native neuronal cells and support their characterization as assembled, alpha3beta4* nAChRs. Although the intracellular nAChRs represent approximately 70% of the total, high-affinity nAChRs expressed in cultured chromaffin cells, they do not appear to be involved in functional recovery after nAChR down regulation. PMID- 12742625 TI - Intraperitoneal corticotropin-releasing factor and urocortin induce Fos expression in brain and spinal autonomic nuclei and long lasting stimulation of colonic motility in rats. AB - CRF injected intraperitoneally (i.p.) stimulates colonic motor function and induces Fos expression in colonic myenteric neurons. We investigated central and spinal Fos expression and changes in colonic motility in response to i.p. injection of CRF and urocortin. Ovine CRF(9-33) that is devoid of intrinsic activity at the CRF receptors, was used as control peptide. Myoelectrical activity was monitored for 1 h before and after peptide injection (10 microg/kg, i.p.) in conscious non fasted rats with chronically implanted intraparietal electrodes in the cecum and proximal colon. Brain and lumbosacral spinal cord were processed for Fos immunohistochemistry at 1 h postinjection. CRF and urocortin elicited defecation and a new pattern of ceco-colonic clustered spike bursts that peaked within 15 min and lasted for the 1 h experimental period while CRF(9-33) did not modify baseline myoelectrical activity and defecation. CRF increased significantly Fos expression in the central nucleus of the amygdala (lateral part), parabrachial nucleus (external lateral subnucleus), area postrema, nucleus tractus solitarius, locus coeruleus, paraventricular nucleus of the hypothalamus, the intermediolateral column and area I-VII, X at the L6-S1 level of the spinal cord by 11-, 6.5-, 5.3-, 5.0-, 4.7-, 2.7- and 1.4-fold, respectively compared with i.p. CRF(9-33) injected rats that had little Fos expression. Urocortin induced a similar pattern of Fos response in the brain and the spinal cord. These results indicate that i.p. CRF and urocortin induce a peptide specific activation of brain nuclei receiving viscerosensory inputs and involved in autonomic circuitries whose effector limbs may impact on visceral function. PMID- 12742626 TI - Reduced serotonin 5-HT1A receptor binding in the temporal cortex correlates with aggressive behavior in Alzheimer disease. AB - Previous studies have implicated brain serotonin 5-HT(1A) receptors in several CNS functions, including cognition, mood and emotional states. In Alzheimer disease (AD), cognitive impairment and behavioral symptoms are the main clinical features. However, the biochemical basis of such changes is poorly understood. Results from recent in vivo studies suggest that 5-HT(1A) receptors may be related to aggressive traits in healthy subjects. The present study investigated the state of 5-HT(1A) receptors in the postmortem neocortex of 33 AD patients prospectively assessed for cognition and behavioral symptoms, together with 20 matched controls, by saturation [(3)H]8-OH-DPAT binding assays. 5-HT(1A) receptor binding affinity (K(D)) and density (B(max)) were unchanged in the overall AD group compared with controls. Within the AD group, 5-HT(1A) receptor B(max) in the temporal cortex inversely correlated with aggression and dementia severity. However, multiple regression analyses showed that 5-HT(1A) receptor B(max) remained the best predictor for aggression, while temporal cortical neurofibrillary tangle grading was the best predictor for dementia severity. This suggests that 5-HT(1A) receptor alteration is directly related to aggression in AD, while dementia severity is more strongly related to the neurodegenerative process. Our data indicate further study of 5-HT(1A) receptors as a pharmacological target for the treatment of behavioral symptoms in AD. PMID- 12742627 TI - Focal cerebral ischemia upregulates SHP-1 in reactive astrocytes in juvenile mice. AB - The role of the tyrosine phosphatase SHP-1 in the hematopoietic system has been well studied; however, its role in the central nervous system (CNS) response to injury is not well understood. Previous studies in our laboratory have demonstrated increased immunoreactivity for SHP-1 in a subset of reactive astrocytes that do not appear to enter the cell cycle following deafferentation of the chicken auditory brainstem. In order to determine whether mammalian astrocytes also upregulate SHP-1 immunoreactivity following CNS injury, a mouse model of focal cerebral ischemia was utilized to study SHP-1 expression. The brains of 3-week-old mice were analyzed at four time points following permanent middle cerebral artery occlusion (MCAO): 1, 3, 7, and 14 days. Our results demonstrate consistent infarct volumes within surgical groups, and infarct volumes decrease as a function of time from 1 day (maximum infarct volume) to 14 days (minimum infarct volume) post-MCAO. In addition, SHP-1 protein levels are upregulated following cerebral ischemia and this increase peaks at 7 days post MCAO. Analysis of confocal images further reveals that immunoreactivity for SHP-1 occurs predominantly in GFAP+ reactive astrocytes, although a small percentage of F4-80+ microglia are also double labeled for SHP-1 at early times post-MCAO. These SHP-1+ reactive astrocytes do not appear to enter the cell cycle (as defined by PCNA immunoreactivity), confirming our previous studies in the avian auditory brainstem. These results suggest that SHP-1 plays an important role in the regulation of glial activation and proliferation in the ischemic CNS. PMID- 12742628 TI - Immunohistochemical study of Ca2+/calmodulin-dependent protein kinase II in the Drosophila brain using a specific monoclonal antibody. AB - To analyze the distribution of Drosophila calcium/calmodulin-dependent protein kinase II (dCaMKII) in the adult brain, we generated monoclonal antibodies against the bacterially expressed 490-amino acid (a.a.) form of dCaMKII. One of those, named #18 antibody, was used for this study. Western blot analysis of the adult head extracts showed that the antibody specifically detects multiple bands between 55 and 60 kDa corresponding to the molecular weights of the splicing isoforms of dCaMKII. Epitope mapping revealed that it was in the region between 199 and 283 a.a. of dCaMKII. Preferential dCaMKII immunoreactivity in the embryonic nervous system, adult thoracic ganglion and gut, and larval neuro muscular junction (NMJ) was consistent with previous observations by in situ hybridization and immunostaining with a polyclonal antibody at the NMJ, indicating that the antibody is applicable to immunohistochemistry. Although dCaMKII immunoreactive signal was low in the retina, it was found at regular intervals in the outer margin of the compound eye. These signals were most likely to be interommatidial bristle mechanosensory neurons. dCaMKII immunoreactivity in the brain was observed in almost all regions and relatively higher staining was found in the neuropilar region than in the cortex. Higher dCaMKII immunoreactivity in the mushroom body (MB) was found in the entire gamma lobe including the heel, and dorsal tips of the alpha and alpha' lobes, while cores of alpha and beta lobes were stained light. Finding abundant dCaMKII accumulation in the gamma lobe suggested that this lobe might especially require high levels of dCaMKII expression to function properly among MB lobes. PMID- 12742629 TI - Blockade of central histaminergic H2 receptors facilitates catecholaminergic metabolism and aggravates ischemic brain damage in the rat telencephalon. AB - Blockade of central H(2) receptors aggravates ischemic neuronal damage. Since changes in the activity of the monoaminergic system are contributing factors in the development of ischemic neuronal damage, the authors evaluated the effects of ranitidine on the monoaminergic system and ischemic neuronal damage in the middle cerebral artery (MCA) occlusion model of rats. Wistar rats pretreated with saline or ranitidine (3 and 30 nmol, i.c.v.) were subjected to reversible occlusion of MCA for 2 h. The total infarct volume was determined 24 h after reperfusion. The relationship between dopaminergic activity and the histologic outcome was estimated by lesioning the substantia nigra 2 days before MCA occlusion. In a second experiment, the animals were subjected to 15 min of MCA occlusion, and the effects of ranitidine on the histologic outcome was evaluated 7 days after ischemia. In a third experiment, the tissue concentrations of monoamines and their metabolites were determined in the cerebral cortex and striatum 2 h after reperfusion following MCA occlusion for 2 h. The turnover of norepinephrine and dopamine was compared between animals treated with saline and those treated with ranitidine by estimating the alpha-methyl-p-tyrosine-induced depletion of norepinephrine and dopamine, respectively. The turnover of 5-hydroxytryptamine was evaluated by the probenecid-induced accumulation of 5-hydroxyindoleacetic acid. Treatments with ranitidine markedly increased the infarct volume 24 h after reperfusion. Ranitidine also aggravated delayed neuronal death 7 days after ischemia. The aggravation was abolished by the lesion of the substantia nigra before MCA occlusion. The MCA occlusion increased the turnover of cortical norepinephrine and striatal dopamine. The turnover was further facilitated by ranitidine. Although ranitidine suppressed the 5-hydroxytryptamine turnover in the cerebral cortex, the extent of this effect was similar in both the ischemic and non-ischemic sides. These results suggest that facilitation of the catecholaminergic systems is involved in the aggravation of ischemic neuronal damage by H(2) blockade. PMID- 12742630 TI - Demonstration and localization of neuronal degeneration in the rat forebrain following a single exposure to MDMA. AB - Methylenedioxymethamphetamine (MDMA, Ecstasy) is a powerful releaser of serotonin. Increasing recreational use of this stimulant and hallucinogenic drug has raised concerns about its potential to produce brain damage. The vast majority of previous research studies have focused on the compound's ability to deplete serotonin (5-hydroxytryptamine, 5-HT) from axon terminals. Despite extensive research on this '5-HT terminal neurotoxicity', a much less studied aspect of MDMA toxicity involves its ability to actually kill nerve cells. Only two prior studies mention the existence of MDMA-induced neuronal degeneration, as reflected by a limited number of argyrophylic neurons within the somatosensory cortex, following very high doses of MDMA. The development of Fluoro-Jade B as a simple and reliable marker of neuronal degeneration has allowed us to conduct the first comprehensive localization of MDMA induced neuronal degeneration throughout the entire rat forebrain. In addition to the previously reported neuronal degeneration within parietal cortex, degenerating neurons were also observed in the insular/perirhinal cortex, the ventromedial/ventrolateral thalamus, and the tenia tecta. The extent of neuronal degeneration observed generally correlated with the degree of hyperthermia achieved. PMID- 12742631 TI - Pathophysiological activity in rat dorsal horn neurones in segments rostral to a chronic spinal cord injury. AB - As a sequel of complete spinal cord injury (SCI), patients often develop chronic pain which is perceived at or just below the level of the lesion. Likewise, in animal models of SCI, spontaneous and evoked pain-related behaviour can be observed. In the present study, the hypothesis was tested that pain related behaviour after SCI in animals is at least partly due to neuronal hyperactivity in spinal segments rostral to the site of injury. In rats with a chronic transected spinal cord, the impulse activity of single dorsal horn neurones was recorded in two locations: (1) directly rostrally adjacent to the lesion, and (2) 2-3 segments more rostrally. Cord transections were made either at the thoracic or at the lumbar level. Sham-operated rats and rats which underwent no surgical interventions served as controls. Compared with both controls, in SCI animals the background activity of the neurones had a significantly higher level in both series. Often the activity showed a pathophysiological altered discharge pattern. Following SCI, there was a general increase in the mechanical responsiveness of neurones that were recorded 2-3 segments rostrally to the lesion. The results suggest that neuronal hyperactivity in spinal segments just rostral to the lesion may contribute to chronic spontaneous SCI pain. Further, there is some indication that the allodynia perceived in body regions near and above the level of the SCI may be due to increased responsiveness to weak stimuli of neurones located more rostrally to the lesion. PMID- 12742633 TI - Increased c-Fos expression in select lateral parabrachial subnuclei following chemical versus electrical stimulation of the dorsal periaqueductal gray in rats. AB - The parabrachial nucleus (PBN) is located in the rostral dorsolateral pons and has been identified as a critical relay for cardiovascular responses (sympathoexcitation and baroreflex attenuation) evoked by the dorsal periaqueductal gray (PAG). We examined the pattern of c-Fos protein immunoreactivity throughout the rostral-caudal extent of the PBN in four groups of anesthetized male Sprague-Dawley rats to identify the specific PBN regions activated by dorsal PAG stimulation. Both electrical stimulation and chemical (0.3 mM bicuculline methobromide) activation of the dorsal PAG elicited a selective increase in Fos-like immunoreactivity (FLI) in the superior lateral and central lateral subnuclei of the rostral lateral PBN (LPBN) relative to surgery and blood pressure control groups. In the middle LPBN chemical stimulation of the dorsal PAG selectively increased FLI in the central lateral subnucleus while electrical stimulation increased FLI in the Kolliker-Fuse area only. Finally, in the caudal LPBN only electrical stimulation of the dorsal PAG induced significant changes in FLI above control. Significant changes in FLI in the medial PBN were not observed under any experimental conditions. These results confirm neuroanatomical data demonstrating that neurons in superior lateral and central lateral subnuclei of the rostral and middle LPBN are the primary targets of the dorsal PAG. Our results also demonstrate that this descending projection to the central lateral and superior lateral subnuclei of the LPBN is in part excitatory. Finally, our results raise the possibility that neurons in the central lateral subnucleus of the middle and rostral LPBN are integrally involved in descending modulation of sympathetic drive associated with dorsal PAG activation. PMID- 12742632 TI - Improgan antinociception does not require neuronal histamine or histamine receptors. AB - Improgan, a chemical congener of the H(2) antagonist cimetidine, induces antinociception following intracerebroventricular (i.c.v.) administration in rodents, but the mechanism of action of this compound remains unknown. Because the chemical structure of improgan closely resembles those of histamine and certain histamine blockers, and because neuronal histamine is known to participate in pain-relieving responses, the antinociceptive actions of improgan were evaluated in mice containing null mutations in the genes for three histamine receptors (H(1), H(2), and H(3)) and also in the gene for histidine decarboxylase (the histamine biosynthetic enzyme). Similar to earlier findings in Swiss-Webster mice, improgan induced maximal, reversible, dose-related reductions in thermal nociceptive responses in ICR mice, but neither pre-improgan (baseline) nor post improgan nociceptive latencies were changed in any of the mutant mice as compared with wild-type controls. Improgan also had weak inhibitory activity in vitro (pK(i)=4.7-4.9) on specific binding to three recently-discovered, recombinant isoforms of the rat H(3) receptor (H(3A), H(3B), and H(3C)). The present findings strongly support the hypothesis that neuronal histamine and its receptors fail to play a role in improgan-induced antinociception. PMID- 12742634 TI - Characterization of nicotinic acetylcholine receptors in cultured arterial chemoreceptor cells of the cat. AB - Neurotransmitters appear to be involved in chemotransmission of the carotid body, a major arterial chemoreceptor. Substantial data indicate that acetylcholine (ACh) is an excitatory neurotransmitter in the carotid body, regulating the excitability of afferent nerve endings and glomus cells (putative chemoreceptor cells). In this study we characterized properties of nicotinic ACh receptors (nAChRs) in cultured cat glomus cells using immunocytochemistry and whole cell patch clamp techniques. Cultured glomus cells expressed immunoreactivity for alpha3, alpha4, and beta2 subunits of nAChRs. An application of ACh elicited inward current. Nicotinic AChRs of glomus cells showed high affinity for ACh. The current-voltage relationship showed strong inward rectification at positive membrane potential. alpha-Conotoxin MII (20 nM), dihydro-beta-erythroidine (DHbetaE; 1 nM), and hexamethonium (300 microM) significantly inhibited ACh induced current. These results indicate that cultured cat glomus cells possess functional nAChRs, and that their characteristics are consistent with those of alpha3, alpha4 and beta2 containing nAChRs. PMID- 12742635 TI - Antigenic compartmentation of the cerebellar cortex in the syrian hamster Mesocricetus auratus. AB - Despite the apparent uniformity in cellular composition of the adult cerebellar cortex, a complex heterogeneous pattern can be revealed by using biochemical markers. One example is zebrin II/aldolase C, which is expressed by a subset of Purkinje cells that form a highly reproducible array of stripes. Zebrin II/aldolase C immunohistochemistry has been used in both section and whole mount preparations to analyze the architecture of the hamster cerebellar cortex. As in other species studied, zebrin II immunoreactivity in the hamster cerebellum is restricted to a subset of Purkinje cells and, more weakly, to astrocytes. Based on the distribution of these Purkinje cell subsets the hamster cerebellar vermis was found to consist of four transverse zones-the anterior zone, central zone, posterior zone and nodular zone. Zebrin II/aldolase C is expressed uniformly in the central and nodular zones, and as parasagittal stripes in the anterior and central zones. A similar alternation of homogeneous and striped expression domains is seen in the hemispheres. The topography of the hamster cerebellar cortex as revealed by zebrin II expression domains closely resembles that reported in other mammals. Thus, a cerebellar zone-and-stripe topography appears to be conserved across the Mammalia. PMID- 12742636 TI - Endogenous proteolytic activity in a rat model of spontaneous cerebral stroke. AB - We evaluated the expression of two extra-cellular protease systems in a model of spontaneous cerebrovascular pathology: spontaneously hypertensive stroke-prone rats (SHRSP). The appearance of brain damage in individual animals was imaged and followed by means of magnetic resonance imaging (MRI). In situ zymography of brain slices obtained 3 days after the appearance of brain damage showed an increase in plasminogen activator (PA)/plasmin activity that co-localised with the cerebral damage detected by MRI; there was also concomitant accumulation/activation of inflammatory cells in the damaged area. Proteolytic activity was inhibited by the urokinase-specific inhibitor amiloride but not by an antibody against tissue-type plasminogen activator (t-PA). SDS-PAGE zymography of brain extracts revealed the presence of 58 kDa plasminogen-dependent lysis areas in the ischemic and non-ischemic tissues, and a 33 kDa lysis area in ischemic tissue only. An antibody against t-PA inhibited the former, whereas the latter was inhibited by amiloride. The specific induction of urokinase-type plasminogen activator (u-PA) in the damaged tissue was further confirmed by the fact that both u-PA protein mass and mRNA were markedly increased in the damaged cerebral areas. Concomitant metalloproteinase-2 (MMP-2) activation was only observed in the damaged area. These data suggest that u-PA is expressed and selectively catalyses proteolysis in the injured area of spontaneous brain damage in SHRSP. PMID- 12742637 TI - Memory enhancing actions of Asiasari radix extracts via activation of insulin receptor and extracellular signal regulated kinase (ERK) I/II in rat hippocampus. AB - Brain insulin receptor and ERK I/II are suggested to play a role in memory formation. We designed a series of experiments to explore if Asiasari radix (AR) extracts could display memory enhancing actions possibly via the activation of insulin receptor and ERK I/II in mice and rats. Methanol extract of AR had significantly increased survival time in the NaNO(2) intoxication assay in mice. Methanol extract of Asiasari radix (fraction 1) and its subfractions, chloroform soluble fraction (fraction 2) and chloroform-insoluble, methanol-soluble fraction (fraction 4) were further tested for memory formation. In eight-arm radial maze experiments, both reference memory errors and working memory errors were significantly decreased in mice by fractions 1, 2 and 4. In addition, these fractions were also effective in promoting memory in the passive avoidance test in mice and rats. To gain insight into the mechanism of memory enhancing effects by Asiasari radix extracts, the activities of hippocampal insulin receptors and ERK I/II were tested in mice and rats. Fraction 1 significantly stimulated tyrosine phosphorylation of the insulin receptor, whereas ERK I/II were stimulated by fractions 1, 2 and 4. These fractions also inhibited cholinesterase activities in rats. These results suggest that Asiasari radix extracts may exert memory enhancing effects via activation of insulin receptor and ERK I/II as well as decreasing cholinesterase activity. PMID- 12742638 TI - Progesterone attenuates the effect of the 5-HT1A receptor agonist, 8-OH-DPAT, and of mild restraint on lordosis behavior. AB - Ovariectomized, hormone-primed rats were used to test the hypothesis that progesterone treatment attenuated the effects of the 5-HT(1A) receptor agonist, (+/-)-8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), on female rat lordosis behavior. Based upon prior evidence that prepriming with estradiol benzoate (EB) reduced the ability of 8-OH-DPAT to inhibit lordosis behavior, rats were preprimed with 10 microg EB 7 days before a second priming with 10 microg EB followed 48 h later with 500 microg progesterone or vehicle. Independent of the presence of progesterone, prepriming with EB attenuated the lordosis-inhibiting effects of systemic treatment with 8-OH-DPAT. However, progesterone also reduced the effects of 8-OH-DPAT and this effect was also seen in females primed only once with EB. In contrast, progesterone was relatively ineffective in attenuating the effects of bilateral infusion with 8-OH-DPAT into the ventromedial nucleus of the hypothalamus (VMN). The failure of progesterone to substantially reduce the effects of VMN infusion with 8-OH-DPAT contrasts with prior studies in which estrogen's protective action against the drug did include the VMN. Thus, while both estrogen and progesterone reduce the lordosis-inhibiting effect of 8-OH DPAT, the mechanisms responsible for the effects of the two gonadal hormones may be different. Priming with progesterone also prevented the effects of 5 min of restraint. When rats were hormonally primed with EB and oil, rats showed a transient, but significant, decline in lordosis behavior 5 and 10 min after restraint. Rats primed with EB and progesterone were unaffected by the restraint. These results are discussed in terms of their implications for the role of progesterone in altering the 5-HT(1A) receptor modulation of lordosis behavior. PMID- 12742640 TI - Kv1.2-immunoreactive primary sensory neurons in the rat trigeminal ganglion. AB - Immunohistochemistry for Kv1.2, a subunit of voltage-gated K(+) channels, was performed on the trigeminal ganglion (TG). Immunoreactivity (ir) was detected in half (48%) the TG neurons. These neurons were mostly medium-sized to large (range 137.6-2664.8 microm(2), mean+/-S.D. 892.6+/-413.3 microm(2)). A double immunofluorescence method also revealed co-expression of Kv1.2 and parvalbumin. Half (54%) the Kv1.2-immunoreactive (ir) neurons exhibited parvalbumin-ir, and parvalbumin-ir neurons mostly showed Kv1.2-ir (95%). Kv1.2-ir neurons which co expressed CGRP-ir were rare in this ganglion. Some 40% of TG neurons retrogradely labeled from the facial skin exhibited Kv1.2-ir, whereas ir was detected in 16% of those labeled from the tooth pulp. The present study indicates that Kv1.2-ir TG neurons include low-threshold mechanoreceptors and nociceptors which innervate the facial skin and tooth pulp, respectively. PMID- 12742639 TI - The novel analgesic and high-efficacy 5-HT1A receptor agonist, F 13640 induces c Fos protein expression in spinal cord dorsal horn neurons. AB - The very-high-efficacy, selective 5-HT(1A) receptor agonist, F 13640 produces uniquely powerful analgesia in rat models of chronic pain by novel neuroadaptive mechanisms (inverse tolerance and co-operation with nociception) [Neuropharmacology 43 (2002) 945-958]. A signal transduction theory and evidence suggest that F 13640 initiates these mechanisms, paradoxically, by mimicking the central effects of nociceptive stimulation. We report that the i.p. injection of F 13640 induces c-Fos protein expression in the L3-L5 segments of the spinal cord. Some 65% of c-Fos protein immunoreactive (c-Fos-IR) nuclei occurred bilaterally in the dorsal horn laminae I-II and V-VI, spinal areas that contain neurons responsive to nociceptive stimulation. This pattern is not unlike that found earlier in arthritic rats, a model of somatotopically widespread nociception. Dose-response studies indicated that c-Fos protein expression was induced at doses (0.63 and 2.5 mg/kg, i.p.) at which previous studies had found F 13640 to produce hyperalgesia. Time-response studies found that c-Fos-IR nuclei appeared within 1-4 h after 0.63 mg/kg of F 13640, with a maximum at 2 h. This parallels literature evidence that c-Fos expression reaches peak late after, and outlasts, nociceptive stimulation. Similar to opioids counteracting noxiously induced c-Fos expression, 10 mg/kg (s.c.) of morphine reduced the number of c-Fos IR nuclei induced by 0.63 mg/kg of F 13640 (by 45+/-5%; P<0.001). The induction by F 13640 of c-Fos protein expression may relate to the initial hyperalgesia which earlier data indicate the agent to produce early upon its administration. PMID- 12742641 TI - Effects of chronic exercise and imipramine on mRNA for BDNF after olfactory bulbectomy in rat. AB - We examined the effects of chronic activity wheel running and antidepressant treatment on brain-derived neurotrophic factor (BDNF) messenger RNA (mRNA) in multiple brain regions-hippocampal formation (HF), ventral tegmental area/substantia nigra (VTA/SN), nucleus accumbens (NAc), and piriform cortex (PFx)-after bilateral olfactory bulbectomy (OBX). Male, Long-Evans rats (n=72) underwent either sham or OBX surgery and were randomly divided into eight experimental groups in a 2 (sham vs. OBX) x 2 (sedentary vs. activity wheel)x2 (saline vs. imipramine) factorial design. Animals were killed after 21 days of treatment. Drug x exercise interaction effects were observed for HF (P=0.006 0.023) and VTA/SN (P=0.021); exercise increased BDNF mRNA in the saline treated animals but not in the imipramine treated animals. OBX did not affect BDNF mRNA in the HF or VTA/SN (P>0.05). BDNF mRNA levels in the PFx were not altered by exercise, drug, or OBX (P>0.05). These results suggest that the effect of exercise on BDNF mRNA extends beyond the HF to the mesolimbic ventral tegmental area and that the potentiation of BDNF mRNA by exercise and antidepressant pharmacotherapy, reported by other investigators, is time limited. PMID- 12742642 TI - Locations of neurons with respiratory-related activity in the ferret brainstem. AB - Previous transneuronal tracing studies conducted in the ferret revealed that a large population of neurons that provides inputs to diaphragm and abdominal motoneurons is located in the ventral magnocellular portion of the medial medullary reticular formation. These observations raise the possibility that the neural substrate underlying respiratory rhythmogenesis may be different in the ferret than in other species in which this circuitry has been explored. In the present study, systematic tracking was conducted through the ferret medulla to map the locations of neurons with activity related to the contractions of respiratory muscles. As in the cat, rat, and rabbit, neurons with respiratory related discharges were distributed either lateral or ventrolateral to the solitary nucleus (dorsal respiratory group) or in the vicinity of nucleus retroambigualis, nucleus ambiguus and the retrofacial nucleus (ventral respiratory group). Although the general organization of respiratory group neurons appeared to be similar in the ferret to that in other mammals, a difference was that few expiratory neurons were located rostrally in the ventral respiratory group. These data suggest that the ventral magnocellular medullary reticular formation is not essential for respiratory rhythm generation, at least during quiet breathing, but may participate in regulating the excitability of respiratory motoneurons or in coordinating the contractions of respiratory muscles during nonrespiratory responses (e.g. coughing or emesis). PMID- 12742643 TI - Involvement of the GABAergic system for Ptychodiscus brevis toxin-induced depression of synaptic transmission elicited in isolated spinal cord from neonatal rats. AB - The involvement of inhibitory transmitters for Ptychodiscus brevis toxin (PbTx) induced depression of spinal synaptic transmission in neonatal rats was investigated. Stimulation of a dorsal root evoked monosynaptic reflex (MSR) and polysynaptic reflex (PSR) potentials in the segmental ventral root. The PbTx depressed the reflexes in a concentration-dependent manner and the depression was blocked by GABA(A) antagonist, bicuculline (1 microM). GABA also produced depression of the reflexes in a concentration-dependent manner. Simultaneous application of submaximal concentrations of PbTx (28 microM) and GABA (30 microM) enhanced the depression (>75%). In contrast, PbTx alone (28 microM) depressed the MSR and the PSR by 33 and 47%, respectively, and GABA (30 microM) alone depressed the reflexes by 30%. The N-methyl-D-aspartate receptor antagonist, DL-2-amino-5 phosphono-pentanoic acid (10 microM), blocked the PbTx-induced depression of MSR and also the enhancement of GABA response by PbTx. A glycine receptor antagonist, strychnine (1 microM), failed to block the depression by the toxin up to 28 microM; however, the depression was attenuated significantly at 84 microM of the toxin. The results indicate that PbTx depressed the spinal reflexes via GABA(A) receptors. Furthermore, the potentiation of GABAergic action by PbTx requires the N-methyl-D-aspartate dependent mechanism. PMID- 12742644 TI - Acute 'binge' cocaine administration elevates dynorphin mRNA in the caudate putamen of C57BL/6J but not 129/J mice. AB - Preprodynorphin mRNA was measured in the nucleus accumbens (NAc) and caudate putamen (CPu) after 3-day 'binge' pattern cocaine administration in C57BL/6J and 129/J mice, strains which differ in behavior and in dopamine increases in the CPu after 'binge' cocaine. In the CPu, there was increased preprodynorphin mRNA in C57BL/6J (P<0.05), but not in 129/J mice, with no differences in the NAc. Thus, 129/J mice are hyporesponsive to the preprodynorphin activating effects of acute 'binge' cocaine in the CPu. PMID- 12742645 TI - Differential sensitivities of mouse strains to morphine and [Dmt1]DALDA analgesia. AB - [Dmt(1)]DALDA (H-Dmt-D-Arg-Phe-Lys-NH(2)), is a highly potent and selective mu opioid agonist. Nevertheless, systemic [Dmt(1)]DALDA retained its analgesic actions in MOR-1 knockout animals and CXBK mice despite the inactivity of morphine in these mice. [Dmt(1)]DALDA was 6-fold less potent in C57BL/6J mice than in CD-1 mice, whereas morphine potency did not differ between the two strains. Thus, [Dmt(1)]DALDA is a highly selective mu-opioid analgesic with significant pharmacological differences with the prototypic mu-opioid morphine. PMID- 12742646 TI - Autoimmune sensorineural hearing loss: an immunologic perspective. AB - Autoimmune sensorineural hearing loss (ASNHL) typically produces a bilateral rapidly progressive loss of hearing that may occur suddenly. The diagnosis is made by excluding ototoxicity, systemic disease, and other factors that mimic ASNHL and by showing a therapeutic response to corticosteroid treatment. Although autoantibodies and autoreactive T cells have been implicated in the etiopathogenesis of ASNHL, several central issues remain unresolved, including the relative prominence of B cell or T cell autoimmunity in the initiation and progression of ASNHL, the identity of the putative inner ear self-antigen(s) that target ASNHL, and the development and application of immunosuppressive therapies for preventing the progressive hearing loss which may be profound and require cochlear implantation. In this review, we will examine the seminal human and animal studies that have led to our current views regarding the autoimmune etiopathogenesis of ASNHL. In addition, we will address the need for developing an inner ear-specific mouse model for ASNHL that may define the stages leading to the development of ASNHL and may also provide new diagnostic markers and help develop novel and effective treatments for preventing progressive hearing loss in ASNHL. PMID- 12742647 TI - 8-[3H]-hydroxy-2-(di-n-propylamino)tetralin binding sites in blood lymphocytes of rats and the modulation by mitogens and immobilization. AB - Serotonin 5-HT(1A) receptors were characterized in rat resting lymphocytes obtained by cardiac puncture with the use of the ligand [3H]8-hydroxy-2-(di-n propylamino)tetralin. Selectivity of the specific binding was demonstrated by inhibition experiments with various serotonergic and nonserotonergic drugs. The rank order of potency for inhibition was WAY-100478>pindobind>NAN 190>buspirone>imipramine>serotonin. While pimozide, desipramine, nomifensine, haloperidol and sulpiride did not inhibit the binding. Kinetic parameters calculated from saturation experiments indicated one site of interaction, with an equilibrium dissociation constant of 2.50 nM and maximum binding capacity of 487.21 nmol/10(6) cells. Complete dissociation was obtained with serotonin as the displacement agent, and equilibrium dissociation constant calculated by association and dissociation experiments was 2.03 nM. Thus, serotonin 5-HT(1A) receptors are present in resting lymphocytes. The in vivo administration of the mitogens lipopolysacharide (0.1 mg/kg, 18 h) or concanavalin A (0.2 mg/kg, 18 h) increased the number of sites. The elevation produced by the latter was of higher magnitude than that of lipopolysacharide, and two sites of the binding were determined by isotopic dilution. Immobilization stress (1 h daily for 7 days) also resulted in a significant increase of binding capacity, but was smaller than that produced by the mitogens. The affinity of binding was not affect by the treatments. The results indicate that serotonin 5-HT(1A) receptors are modulated by unspecific and specific immune system activation, as well as by a potent stress condition, which might result in relevant functional modifications in the response of rat lymphocytes. PMID- 12742648 TI - Social stress affects migration of blood T cells into lymphoid organs. AB - The effect of social confrontation stress on the normal distribution of blood T cells into lymphoid organs and some other body tissues was studied. Social stress was induced by placing a male Fischer 344 (F334) rat into the home cage of a resident opponent. 51Cr-labeled blood T cells isolated from syngeneic donor, were intravenously injected into recipients immediately before confrontation. The accumulation of 51Cr-labeled T cells in the spleen, mesenteric and cervical lymph nodes 24 h after injection was about 30% lower in socially stressed than in control males. Substantially higher localization was observed in the bone marrow of socially stressed males. PMID- 12742649 TI - Polyclonal IgM influence oligodendrocyte precursor cells in mixed glial cell cultures: implications for remyelination. AB - Polyclonal immunoglobulins for intravenous use (IVIg) are a potent immunomodulator and have been shown to be effective in several immune-mediated diseases. This includes inflammatory demyelinating diseases of the central nervous system (CNS) like multiple sclerosis (MS). Besides their immunomodulatory function, IVIg have been proposed to enhance remyelination based on studies in the animal model of Theiler's murine encephalomyelitis virus (TMEV). Disappointingly, recent treatment trials in patients with MS have failed to demonstrate repair of longstanding deficits. Since the clinical trials have used IVIg that contained nearly exclusively IgG, whereas the most pronounced effect in TMEV was seen with IgM, this could be a possible explanation for the negative outcome in the MS trials. Here we have examined the effects of a new polyclonal IgM preparation (IVIgM) on cultured oligodendrocyte precursor cells (OPCs). To achieve successful remyelination, OPCs proliferate, migrate, and differentiate into mature myelinating oligodendrocytes. IVIgM and commercial IVIg preparations had no influence on proliferation and differentiation of either isolated OPCs or OPCs in coculture with microglia. In contrast, IVIgM inhibited the proliferation of OPCs in mixed glial cultures containing astrocytes and microglia. This was not seen in cultures treated with IVIg, albumin, or interferon-gamma (IFN-gamma), suggesting that this is a specific effect of IVIgM. Differentiation was slightly delayed by IVIgM in mixed glial cultures, but this was not statistically significant and interferon-gamma had a similar effect. These results underline the importance of IgM in influencing OPCs and corroborate the in vivo findings that polyclonal IgM are more potent than IgG in their capacity to influence remyelination. The exact mechanism of how this modulation of OPCs is achieved remains unknown, but a complex interaction among all cells present in the CNS has to be postulated. PMID- 12742650 TI - Interleukin-18 gene-deficient mice show enhanced defense and reduced inflammation during pneumococcal meningitis. AB - To determine the role of endogenous interleukin-18 (IL-18) in pneumococcal meningitis, meningitis was induced in IL-18 gene-deficient (IL-18(-/-)) and wild type (WT) mice by intranasal inoculation of Streptococcus pneumoniae with hyaluronidase. Induction of meningitis resulted in an upregulation of both pro- and mature IL-18 in brain tissue in WT mice. IL-18(-/-) and WT mice were equally susceptible to develop meningitis after intranasal infection, yet IL-18(-/-) mice showed a prolonged survival and a suppressed inflammatory response, as reflected by a less profound inflammatory infiltrate around the meninges and lower concentrations of cytokines and chemokines in brain tissue. These findings suggest that endogenous IL-18 contributes to a detrimental inflammatory response during pneumococcal meningitis and that elimination of IL-18 may improve the outcome of this disease. PMID- 12742651 TI - Somatostatin binds to murine macrophages through two distinct subsets of receptors. AB - Somatostatin (SRIF) exerts anti-inflammatory effects, in part by deactivating monocytes/macrophages. Thus, the objective of this study was to characterize specific receptors for SRIF on these cells. Macrophages isolated from mouse peritoneal cells bound [125I]Tyr(0), D-Trp(8) SRIF(14) specifically. Scatchard analysis of saturation binding data revealed two classes of binding sites with an affinity of 0.44+/-0.13 and 2.58+/-0.56 nM, respectively. By sensitive and specific RT-PCR, the mRNAs for the five SRIF receptors (SSTR1 to SSTR5) could be detected. Evidence for the involvement of SSTR1 and SSTR2 in the binding of SRIF to the high and low affinity sites, respectively, was obtained by the demonstration that (1) only SSTR1 and SSTR2 subtype-specific agonists were active in competing for [125I]Tyr(0), D-Trp(8) SRIF(14) binding to high and low affinity sites, respectively, and (2) [125I]Tyr(0), D-Trp(8) SRIF(14) bound to high but not low affinity sites on macrophages isolated from SSTR2 knock-out mice. In conclusion, we have identified and characterized two different SRIF receptor subtypes in murine macrophages. PMID- 12742652 TI - Quantification of neurodegeneration by measurement of brain-specific proteins. AB - Quantification of neurodegeneration in animal models is typically assessed by time-consuming and observer-dependent immunocytochemistry. This study aimed to investigate if newly developed ELISA techniques could provide an observer independent, cost-effective and time-saving tool for this purpose. Neurofilament heavy chain (NfH(SM135)), astrocytic glial fibrillary acidic protein (GFAP), S100B and ferritin, markers of axonal loss, gliosis, astrocyte activation and microglial activation, respectively, were quantified in the spinal cord homogenates of mice with chronic relapsing experimental allergic encephalomyelitis (CREAE, n=8) and controls (n=7). Levels of GFAP were found to be threefold elevated in CREAE (13 ng/mg protein) when compared to control animals (4.5 ng/mg protein, p<0.001). The inverse was observed for NfH(SM135) (21 ng/mg protein vs. 63 ng/mg protein, p<0.001), ferritin (542 ng/mg protein vs. 858 ng/mg protein, p<0.001) and S100B (786 ng/mg protein vs. 2080 ng/mg protein, N.S.). These findings were confirmed by immunocytochemistry, which demonstrated intense staining for GFAP and decreased staining for NfH(SM135) in CREAE compared to control animals. These findings indicate that axonal loss and gliosis can be estimated biochemically using the newly developed ELISA assays for NfH(SM135) and GFAP. These assays may facilitate the quantification of pathological features involved in neurodegeneration. PMID- 12742653 TI - Maternal infection regulates BDNF and NGF expression in fetal and neonatal brain and maternal-fetal unit of the rat. AB - Maternal infection during pregnancy is associated with increased risk for neurodevelopmental disorders. Lipopolysaccharide (LPS) or saline was administered to rats to model maternal infection, and levels of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in maternal plasma, placenta, amniotic fluid, fetal liver/spleen, fetal brain, and cerebral cortex after birth were determined by ELISA or semiquantitative Western blot analysis. BDNF expression was significantly increased in the fetal brain (p=0.039); NGF expression was significantly increased in neonatal cortex (p=0.0009). Neurotrophic factor expression was also altered in other tissues of the maternal fetal unit. Abnormal expression of neurotrophic factors represents a potential mechanism through which maternal infection increases risk for neurodevelopmental disorders. PMID- 12742654 TI - A novel nitric oxide scavenger in combination with cyclosporine A ameliorates experimental autoimmune encephalomyelitis progression in mice. AB - Immunotherapy improves experimental autoimmune encephalomyelitis (EAE), a model of multiple sclerosis (MS), while excessive production of nitric oxide (NO) has been implicated in the pathogenesis of this disease. Here, we show that disease progression in SJL/J mice with EAE is improved after treatment with either a subtherapeutic dose of cyclosporine A (CsA) or NOX-100, a nitric oxide scavenger. Importantly, the impact of subtherapeutic doses of CsA in combination with NOX 100 on disease progression in EAE was greater than that attained with either agent alone and led to near total protection. CNS inflammation and gene expression of proinflammatory cytokines and iNOS were also significantly reduced after treatment. These observations point to the potential therapeutic utility of NOX-100 as a dose-reducing agent for CsA in the treatment of MS. PMID- 12742655 TI - Crosstalk between neurokinin receptors is relevant to hematopoietic regulation: cloning and characterization of neurokinin-2 promoter. AB - Neurokinin (NK)-1 and NK-2 receptors regulate hematopoiesis by interacting with neurotransmitters that belong to the tachykinin. This report studies the relationship between NK-1 and NK-2 in primary human bone marrow (BM) stroma, which supports hematopoiesis. Use of NK receptor antagonists and deficient stromal cells indicate that the neurotransmitter, substance P (SP), could exert dual hematopoietic effects (inhibitory or stimulatory), depending on the interacting receptor and crosstalk between NK-1 and NK-2. Cloning and identification of the minimal promoter for NK-2 and comparison with NK-1 promoter showed that the hematopoietic functions of NK receptors involve receptor crosstalk and the particular cytokine (IL-3, GM-CSF, TGF-beta or IL-1alpha). Crosstalk between NK-1 and NK-2 adds to communication within neural-hematopoietic axis. PMID- 12742656 TI - Decreased expression of Src homology 2 domain-containing protein tyrosine phosphatase 1 reduces T cell activation threshold but not the severity of experimental autoimmune myasthenia gravis. AB - Myasthenia gravis (MG) and its murine model experimental autoimmune myasthenia gravis (EAMG) are T cell-dependent, antibody-mediated autoimmune diseases. Src homology 2 domain-containing protein tyrosine phosphatase 1 (SHP-1) is a cytosolic tyrosine phosphatase that is involved in regulating the T cell activation cascade from signals initiated through the TCR. To study the role of SHP-1 in EAMG pathogenesis, we immunized C57BL/6 (B6) mice heterozygous for deletion of the SHP-1 gene (me(v+/-)) and their littermate wild type B6 mice with torpedo acetylcholine receptor (TAChR). T cell proliferation and IFNgamma production were significantly increased in B6.me(v+/-) mice after immunization with AChR compared to that of wild type littermates. However, clinical incidence and severity of the disease were not changed. There also were no significant differences in AChR-specific antibodies produced between wild type and me(v+/-) mice. These data suggest that deficiency in SHP-1 expression does decrease the activation threshold of autoreactive T cells in EAMG, but the increased frequency of autoreactive T cells does not aggravate EAMG in terms of clinical score, incidence, or antibody titers. PMID- 12742657 TI - Glucocorticoid involvement in suppression of NK activity following surgery in rats. AB - We studied plasma factors mediating suppression of NK activity (NKA) following surgery. Plasma from operated rats suppressed NKA of splenocytes, leukocytes, and purified natural killer (NK) cells, and charcoal stripping nullified suppression. The glucocorticoid antagonist mifepristone prevented suppression, whereas blockers of reactive oxygen metabolites, opioids, catecholamines, prostaglandin E2, and histamine did not. NKA dropped as corticosterone levels peaked postoperatively, and administration of relevant doses of corticosterone suppressed NKA. Inhibition of glucocorticoid synthesis prevented plasma from suppressing NKA but merely attenuated NKA suppression in operated rats. Thus, postoperative concentrations of corticosterone can directly suppress NKA but additional factors probably act in vivo. PMID- 12742659 TI - Basal serum levels and reactivity of nerve growth factor and brain-derived neurotrophic factor to standardized acute exercise in multiple sclerosis and controls. AB - Neurotrophins like brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) are thought to play an important role in neuronal repair and plasticity. Recent experimental evidence suggests neuroprotective effects of these proteins in multiple sclerosis (MS). We investigated the response of serum NGF and BDNF concentrations to standardized acute exercise in MS patients and controls. Basal NGF levels were significantly elevated in MS. Thirty minutes of moderate exercise significantly induced BDNF production in MS patients and controls, but no differential effects were seen. We conclude that moderate exercise can be used to induce neutrophin production in humans. This may mediate beneficial effects of physical exercise in MS reported recently. PMID- 12742658 TI - Intraventricular infusion of antagonists of IL-1 and TNF alpha attenuates neurodegeneration induced by the infection of Trypanosoma brucei. AB - Infection of Trypanosoma brucei causes specific patterns of neurodegeneration in association with chronic expression of proinflammatory cytokines in the brain. To investigate whether the induction of proinflammatory cytokines contributed to the observe pathology in this disease, we infected rats with T. brucei and treated them with intracerebral infusion of the cytokine antagonists interleukin-1 receptor antagonist (IL-1ra) and/or soluble type-I receptor of the tumor necrosis factor (sTNFr1). Infusion of IL-1ra, not sTNFr1, restored the reduction of body weight gain induced by the infection. Infusion of IL-1ra+sTNFr1 reduced the expression of IL-1beta and the cytokine response gene IkappaBalpha, but not TNFalpha. Infusion of sTNFr1 reduced trypanosome-induced neurodegeneration. Further reduction of neurodegeneration was seen after IL-1ra+sTNFr1 infusion. Infusion of IL-1ra alone, however, did not significantly affect the patterns of neurodegeneration. These results suggest that TNFalpha is a major mediator for trypanosome-induced neurodegeneration although its neurotoxic effects can be augmented by IL-1. PMID- 12742660 TI - Matrix metalloproteinase-12 is expressed in phagocytotic macrophages in active multiple sclerosis lesions. AB - Matrix metalloproteinases (MMPs) are proteases involved in extracellular matrix (ECM) remodeling, leukocyte infiltration into lesions and myelin degradation in the central nervous system (CNS) disease multiple sclerosis (MS). We have investigated whether MMP-12 (macrophage metalloelastase) is expressed in MS lesions at various stages. In control patient tissue and (p)reactive MS lesions, only occasional microglial and astrocyte staining was detected. In contrast, in active demyelinating lesions, phagocytic macrophages were MMP-12 positive. A lower proportion of phagocytes was positive for MMP-12 in chronic active demyelinating lesions and inactive lesions. This suggests a role for MMP-12 during demyelination in MS. PMID- 12742661 TI - Tryptase activates peripheral blood mononuclear cells causing the synthesis and release of TNF-alpha, IL-6 and IL-1 beta: possible relevance to multiple sclerosis. AB - Presence of mast cells and an increase in the concentration of their products has been reported in multiple sclerosis (MS) plaques. The most abundant secretory mediator of the human mast cell is the tetrameric protease tryptase. We demonstrate that tryptase can activate peripheral mononuclear cells (PBMCs), isolated from healthy donors as well as MS patients for the release of tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-6 and IL-1beta. Cytokine secretion was significantly higher in secondary progressive (SP) MS patients and healthy control (HC) individuals than in relapsing-remitting (RR) patients. Our findings suggest that tryptase is, most probably, an important mediator of inflammation in MS. PMID- 12742662 TI - The development of clinical activity in relapsing-remitting MS is associated with a decrease of FasL mRNA and an increase of Fas mRNA in peripheral blood. AB - In this longitudinal study, we examined the expression of Fas, FasL, CCR3, CCR5 and CXCR3 mRNA in peripheral blood mononuclear cells (PBMCs) of secondary progressive (SP) and relapsing-remitting (RR) multiple sclerosis (MS) patients. In RR patients, FasL, CCR3 and CCR5 mRNA levels were increased prior to the exacerbations, but these decreased during clinical activity, while mRNA levels of Fas increased. SP patients have increased the levels of Fas and FasL mRNA; the latter was particularly increased during lesional activity. Our data support the hypothesis that changes in Fas and FasL mRNA related to clinical activity are due to the migration of inflammatory cells to the central nervous system (CNS). PMID- 12742663 TI - Antigen processing and presentation in human muscle: cathepsin S is critical for MHC class II expression and upregulated in inflammatory myopathies. AB - The immunological properties of muscle cells are of critical importance for both the pathogenesis of inflammatory muscle disorders as well as for understanding and controlling novel therapeutic strategies. Muscle cells can present antigens to both CD4 and CD8 cells. However, the cellular biochemistry of antigen processing and presentation by muscle cells is not clear. Cathepsins play a central role in the generation of antigenic peptide and control transport and maturation of MHC class II molecules. To further elucidate the molecular basis for the MHC class II-mediated antigen presentation by muscle cells, we here analyzed cultured human myoblasts and biopsies from inflammatory myopathies with respect to the expression and function of the constituents of the MHC class II antigen presentation machinery. We identified cathepsin S (CatS) as the dominant endocytic protease that is specifically upregulated under inflammatory conditions to significant mRNA levels, synchronously with HLA-DR, -DM and the class II invariant chain (Ii), both in muscle biopsies from affected individuals with inflammatory myopathies and in human myoblasts cultured in the presence of IFN gamma. This led to translation of the mature CatS polypeptide that was enzymatically active in human myoblasts under inflammatory conditions. By contrast, expression of CatL and CatB was unaffected by IFN-gamma at both the expression and activity levels. CatS activity is required for efficient surface display of MHC class II in this cell type: functional inhibition of CatS using a CatS-selective inhibitor reduced the levels of surface class II alphabeta:peptide complexes on stimulated myoblasts by almost 50%. Surprisingly, and in contrast to B cells and dendritic cells, this was not due to inefficient processing of Ii in the absence of CatS, which was unaffected by the elimination of CatS activity. We therefore conclude that CatS is involved in the regulation of class II expression in human myoblasts independently from Ii processing. PMID- 12742664 TI - Neuronal injury regulates fractalkine: relevance for HIV-1 associated dementia. AB - Fractalkine (FKN), a chemokine highly expressed in the central nervous system, participates in inflammatory responses operative in many brain disorders including HIV-1 associated dementia (HAD). In this report, HIV-1 progeny virions and pro-inflammatory products led to FKN production associated with neuronal injury and apoptosis. FKN was produced by neurons and astrocytes; but differentially produced by the two cell types. Laboratory tests paralleled those in infected people where cerebrospinal fluid FKN levels in HIV-1 infected cognitively impaired (n=16) patients were found to be increased when compared to infected patients without cognitive impairment (n=8, P=0.0345). These results demonstrate a possible role of FKN in HAD pathogenesis. PMID- 12742665 TI - Autoimmune markers in HIV-associated dementia. AB - The etiology of HIV-associated dementia (HAD) is still unknown although direct viral effects have not been supported. Although evidence supports a role for products of activated macrophages, other evidence suggested the possibility of associated autoimmune phenomena at least as a marker. In a blinded analysis, non HIV-infected whole brain material was immunoblotted with samples of serum, and in certain cases cerebrospinal fluid (CSF), from HAD patients and controls. Distinct antibrain antibodies were detected in 11/12 of HIV+ HAD patients, 7/19 of HIV+ patients without HAD, and 0/11 HIV seronegative controls who were either healthy or had other neurologic diseases. Reactivity against control tissue was negative. Though the etiopathogenetic relation of these antibrain antibodies remains to be delineated, the data suggest that they may be a marker of HAD. PMID- 12742666 TI - Working memory: an introduction to the conference proceedings. PMID- 12742667 TI - Working memory and language: an overview. AB - Working memory involves the temporary storage and manipulation of information that is assumed to be necessary for a wide range of complex cognitive activities. In 1974, Baddeley and Hitch proposed that it could be divided into three subsystems, one concerned with verbal and acoustic information, the phonological loop, a second, the visuospatial sketchpad providing its visual equivalent, while both are dependent upon a third attentionally-limited control system, the central executive. A fourth subsystem, the episodic buffer, has recently been proposed. These are described in turn, with particular reference to implications for both the normal processing of language, and its potential disorders. The reader will be introduced to the concept of a multi-component working memory. Particular emphasis will be placed on the phonological loop component, and (a) its fractionation into a storage and processing component, (b) the neuropsychological evidence for this distinction, and (c) its implication for both native and second language learning. This will be followed by (d) a brief overview of the visuospatial sketchpad and its possible role in language, culminating in (e) discussion of the higher-level control functions of working memory which include (f) the central executive and its multi-dimensional storage system, the episodic buffer. An attempt throughout is made to link the model to its role in both normal and disordered language functions. PMID- 12742668 TI - Effects of working memory deficits on the communicative functioning of Alzheimer's dementia patients. AB - Individuals with Alzheimer's disease experience frontal lobe pathology and deficits in working memory processes are well documented. Less documented is how various working memory deficits impact communicative functioning. The performance data of individuals with mild and moderate Alzheimer's dementia on five tests of language comprehension and four tests of language expression are presented and discussed in the context of possible contributions from impaired working memory functions. The argument is advanced that diminished scores on tests of language comprehension and production result primarily from attenuated span capacity, difficulty focusing attention, encoding, and activation of long-term knowledge rather than from loss of linguistic knowledge. Techniques that may advantage Alzheimer's patients in the comprehension and expression of linguistic information are discussed.(1) Readers will become familiar with the typical functioning of individuals with Alzheimer's disease on common linguistic expression and comprehension tasks. (2) Readers will become familiar with the distinction between language knowledge and performance and its importance in understanding the cause of communication breakdowns in individuals with Alzheimer's disease. (3) Readers will become familiar with techniques that may facilitate the communicative functioning of individuals with Alzheimer's disease. PMID- 12742669 TI - Working memory and comprehension in children with specific language impairment: what we know so far. AB - Many children with specific language impairments (SLI) demonstrate deficits in the areas of verbal working memory and language learning/processing. In this article, evidence is reviewed suggesting that the lexical/morphological learning and sentence comprehension/processing problems of many of these children are associated with their deficient working memory functioning. Evidence is also reviewed for the possibility that deficient working memory provides a clinical marker of SLI. A number of potentially useful assessment and intervention techniques are offered, as well as several directions for future research. The reader will be introduced to two prominent models of verbal working memory (phonological working memory model, functional working memory) and how each model potentially relates to (a) various language abilities in typically developing children, (b) the morphological and lexical learning abilities in children with specific language impairment (SLI), and (c) the sentence comprehension of children with SLI. The reader will also be provided a variety of clinical suggestions on how to assess and treat the working memory and language processing problems of children with SLI. Finally, some suggestions for future research will also be offered. PMID- 12742670 TI - Language representation and working memory with bilinguals. AB - Working memory (WM) plays a crucial role in learning a second language (L2). The ability to repeat words in an unknown language has been observed to predict success in learning that language. Conversely, decreased digit span and inability to repeat pseudowords have been related with failure in L2 acquisition. Not only digit span, but also "word span" and "semantic span" should be considered in WM analysis. In addition to the phonological system, a semantic system is proposed in the WM model for language. In bilinguals, brain activation patterns during WM tasks have been observed to be more complex when using a L2. Processing information in L2 is more demanding, and WM may be less efficient. It can be conjectured that language understanding defects in L2 are at least partially due to this decreased efficiency of WM in its phonological as well as in its semantic subsystem. The reader will be introduced to the basic assumptions of WM. It will be emphasized that WM is significantly involved in the ability to learn a L2. Cross-linguistic differences in digit span will be analyzed. It will be concluded that despite digit span and word span are affected by different variables, "semantic span" may be similar across languages. Words in a L2 function as low frequency words, and hence, semantic search takes longer and WM is less efficient. It will be concluded that in addition to the "phonological system," the WM model should include a "semantic system," involving a "semantic store" and a "semantic search" process. PMID- 12742671 TI - Modern proteomic strategies in the study of complex neuropsychiatric disorders. AB - An insight into protein mechanisms involved in disease is critical to the discovery and design of new therapeutic tools. Direct protein analysis provides a method for studying the proteome of a tissue irrespective of an in-depth knowledge of its transcriptome. The development of a human central nervous system (CNS) proteome database ultimately will serve to accelerate the development of specific diagnostic and prognostic markers, neuropsychiatric disease markers, and the corresponding therapeutic tools. It may also reduce the uncertainties in in silico gene predictions by direct open reading frame verification and the ambiguities that experimental models of disease may provide. Advances in gel independent proteomic analyses by solid phase isotope tagging provide greater scope for the characterization of previously elusive membrane proteins; approximately half of all drug targets are key CNS membrane proteins. These advances hold great promise for improvements in the understanding, diagnosis, and therapy of central nervous system disorders. PMID- 12742672 TI - Differential amygdalar response to novel versus newly familiar neutral faces: a functional MRI probe developed for studying inhibited temperament. AB - BACKGROUND: As a prelude to future studies of subjects with different temperaments, we sought to develop a probe to measure differential amygdalar responses to novel versus familiar stimuli. Prior neuroimaging studies of the amygdala in humans to date have focused principally on responses to emotional stimuli, primarily aversive, rather than to novelty per se. METHODS: Eight normal subjects aged 22.4 +/- 1.3 years were scanned using functional magnetic resonance imaging (fMRI) during passive viewing of novel and familiar faces. RESULTS: Using this newly developed paradigm, we found greater fMRI blood oxygenation level dependent (BOLD) signal response within the right amygdala to novel versus familiar faces--all with neutral expression. Furthermore, although a new facial identity was always presented in the novel condition, signal in the amygdala declined over time as it did for the familiar condition. CONCLUSIONS: These results suggest that at least one primary function of the amygdala is to detect and process unexpected or unfamiliar events that have potential biological import, of which stimuli symbolic of fear or threat are but one possible example. We propose that this experimental paradigm will be useful for examining brain responses to novelty in different temperamental groups, as well as various psychiatric disorders. PMID- 12742673 TI - Rodent doxapram model of panic: behavioral effects and c-Fos immunoreactivity in the amygdala. AB - BACKGROUND: Panic attacks, the hallmark of panic disorder, are often characterized by hyperventilation. Existing animal models of anxiety have not addressed the effects of the hyperventilation on anxiety-related behaviors. Doxapram is a respiratory stimulant that reliably evokes panic attacks in patients with panic disorder. We examined doxapram in four rodent models of anxiety and sought to identify brain regions involved in its behavioral effects. METHODS: The effects of doxapram were determined for cue and contextual fear conditioning, the open field test, and the social interaction test. The effect of doxapram on c-Fos-like immunoreactivity was examined in three brain regions. RESULTS: Doxapram at 4 mg/kg increased anxiety-related behaviors in all four anxiety models. An inverted U-shaped dose-response curve was identified for fear conditioning to cue. Doxapram induced c-Fos-like immunoreactivity in the central nucleus of the amygdala but not the lateral nucleus or the nucleus tractus solitarius. CONCLUSIONS: Doxapram enhanced anxiety-related behaviors in four animal models of anxiety that involve conditioning or spontaneous avoidance. The effect of doxapram may result from activation of neurons in the amygdala. Doxapram, by inducing hyperventilation, may be a useful adjunct to existing animal anxiety models for improving validity for panic anxiety. PMID- 12742674 TI - Differential patterns of striatal activation in young children with and without ADHD. AB - BACKGROUND: Cognitive control, defined as the ability to suppress inappropriate thoughts and actions, is compromised in attention-deficit/hyperactivity disorder (ADHD). This study examines the neural basis of this deficit. METHODS: We used a paradigm that incorporates a parametric manipulation within a go/nogo task, so that the number of go trials preceding a nogo trial is varied to tax the neural systems underlying cognitive control with increasing levels of interference. RESULTS: Using this paradigm in combination with event-related functional magnetic resonance imaging (fMRI), we show that children without ADHD have increased susceptibility to interference with increasing numbers of go trials preceding a nogo trial, but children with ADHD have difficulty even with a single go trial preceding a nogo trial. In addition, children with ADHD do not activate frontostriatal regions in the same manner as normally developing children, but rather rely on a more diffuse network of regions, including more posterior and dorsolateral prefrontal regions. CONCLUSIONS: Normal immature cognition may be characterized as being susceptible to interference and supported by the maturation of frontostriatal circuitry. ADHD children show a slightly different cognitive profile at 6 to 10 years of age that is paralleled by a relative lack of or delay in the maturation of ventral frontostriatal circuitry. PMID- 12742675 TI - Neural correlates of declarative memory for emotionally valenced words in women with posttraumatic stress disorder related to early childhood sexual abuse. AB - BACKGROUND: Animal studies have shown that early stressors result in lasting changes in structure and function of brain areas involved in memory, including hippocampus and frontal cortex. Patients with childhood abuse-related posttraumatic stress disorder (PTSD) have alterations in both declarative and nondeclarative memory function, and imaging studies in PTSD have demonstrated changes in function during stimulation of trauma-specific memories in hippocampus, medial prefrontal cortex, and cingulate. The purpose of this study was to assess neural correlates of emotionally valenced declarative memory in women with early childhood sexual abuse and PTSD. METHODS: Women with early childhood sexual abuse-related PTSD (n = 10) and women without abuse or PTSD (n = 11) underwent positron emission tomographic (PET) measurement of cerebral blood flow during a control condition and during retrieval of neutral (e.g., "metal iron") and emotionally valenced (e.g., "rape-mutilate") word pairs. RESULTS: During retrieval of emotionally valenced word pairs, PTSD patients showed greater decreases in blood flow in an extensive area, which included orbitofrontal cortex, anterior cingulate, and medial prefrontal cortex (Brodmann's areas 25, 32, 9), left hippocampus, and fusiform gyrus/inferior temporal gyrus, with increased activation in posterior cingulate, left inferior parietal cortex, left middle frontal gyrus, and visual association and motor cortex. There were no differences in patterns of brain activation during retrieval of neutral word pairs between patients and control subjects. CONCLUSIONS: These findings are consistent with dysfunction of specific brain areas involved in memory and emotion in PTSD. Regions implicated in this study of emotionally valenced declarative memory are similar to those from prior imaging studies in PTSD using trauma-specific stimuli for symptom provocation, adding further supportive evidence for a dysfunctional network of brain areas involved in memory, including hippocampus, medial prefrontal cortex, and cingulate, in PTSD. PMID- 12742676 TI - The efficacy of reboxetine in preventing and reverting a condition of escape deficit in rats. AB - BACKGROUND: Different stress-induced experimental models of depression are currently used to study the efficacy and mechanism of action of classical or potential antidepressant compounds. We studied the effect of single and repeated administrations of reboxetine, an antidepressant that selectively inhibits noradrenaline reuptake, in the prevention and reversal of stress-induced escape deficit. Moreover, we examined the effect of chronic reboxetine on the stress induced decrease of dopamine output in the nucleus accumbens shell (NAcS). METHODS: Rats received a single or 21-day reboxetine administration before acute unavoidable stress exposure; 24 hours later their escape response was examined. Rats were exposed to repeated unavoidable stress for 21 days, with or without reboxetine treatment, and then were tested for escape; 2 days later they were implanted with microdialysis probes in the NAcS. RESULTS: A single reboxetine administration showed a protective activity on stress-induced escape deficit development that significantly increased after 21 days of treatment. This effect was antagonized by propranolol, a selective beta-adrenergic antagonist. In rats exposed to chronic stress, a 21-day reboxetine treatment reinstated the avoidance response and NAcS dopamine output to control values. CONCLUSIONS: In these stress induced models, long-term reboxetine administration showed a protective activity similar to that of classical antidepressants. PMID- 12742677 TI - Thyroid hypofunction in patients with rapid-cycling bipolar disorder after lithium challenge. AB - BACKGROUND: There is debate whether patients with rapid-cycling bipolar disorder (BD) are predisposed to thyroid axis abnormalities and whether this may contribute to development of rapid mood shifts. Using lithium carbonate as a challenge to the hypothalamic-pituitary-thyroid (HPT) system, we determined whether patients with rapid-cycling BD are sensitive to the "antithyroid" properties of lithium. METHODS: We studied the response to thyrotropin-releasing hormone (TRH) of HPT system hormones in 20 medication-free patients with rapid cycling BD and compared these measurements with those of 20 healthy age- and gender-matched control subjects. The same measurements were repeated after both groups had received lithium carbonate for 4 weeks in sufficient doses to maintain blood levels between.7-1.2 mEq/L. RESULTS: At baseline, the results of thyroid function tests, including the TRH challenge test, did not differ between patients and control subjects. After treatment with lithium, serum concentrations of thyroxine significantly decreased, whereas basal thyrotropin (TSH) and DeltaTSH(max) significantly increased in both patients and control subjects; however, patients had significantly higher DeltaTSH(max) after TRH stimulation. More patients than control subjects developed laboratory evidence consistent with grade III hypothyroidism after lithium treatment. CONCLUSIONS: Rapid-cycling BD is associated with a latent hypofunction of the HPT system. This dysfunction becomes manifest with short-term lithium challenge. PMID- 12742678 TI - Neuronal pathology in the hippocampal area of patients with bipolar disorder: a study with proton magnetic resonance spectroscopic imaging. AB - BACKGROUND: The brain regions involved in the pathophysiology of bipolar disorder have not been definitively determined. Previous studies have suggested possible involvement of the hippocampus and of prefrontal regions. Proton magnetic resonance spectroscopic imaging ((1)H-MRSI) allows measurement of N acetylaspartate (NAA, marker of neuronal integrity), choline-containing compounds (CHO), and creatine+phosphocreatine (CRE) in multiple brain regions. The objective of this study was to assess possible NAA reductions in hippocampus and prefrontal regions in patients with bipolar disorder. METHODS: We studied 17 patients with bipolar disorder and 17 age- and gender-matched healthy subjects on a 1.5-T nuclear magnetic resonance (NMR) machine. With (1)H-MRSI we measured ratios of areas under the metabolite peaks of the proton spectra (i.e., NAA/CRE, NAA/CHO, CHO/CRE) for multiple cortical and subcortical regions. RESULTS: Patients showed significant reductions of NAA/CRE bilaterally in the hippocampus. There were no significant changes in CHO/CRE or in NAA ratios in any other area sampled. CONCLUSIONS: This study shows that patients with bipolar disorder have a regional reduction of NAA relative signals, suggesting neuronal damage or malfunction of the hippocampus. As suggested by other studies, neuronal pathology in the hippocampus may be involved in the pathophysiology of bipolar disorder and in susceptibility to psychosis. PMID- 12742679 TI - The identification of OCD-related subgroups based on comorbidity. AB - BACKGROUND: Individuals with obsessive-compulsive disorder (OCD) frequently have other psychiatric disorders. This study employed latent class analysis (LCA) to explore whether there are underlying clinical constructs that distinguish "OCD related" subgroups. METHODS: The study included 450 subjects, case and control probands and their first-degree relatives, and LCA was used to derive empirically based subgroups of 10 disorders: OCD, obsessive-compulsive personality disorder (OCPD), recurrent major depressive disorder (RMDD), separation anxiety disorder, panic disorder or agoraphobia (PD/AG), tic disorders (TD), generalized anxiety disorder (GAD), somatoform disorders (hypochondriasis or body dysmorphic disorder), pathologic skin picking or nail biting (PSP/NB), and eating disorders (EDs). The derived classes were compared on several clinical variables. RESULTS: The best fitting model is a four-class structure: minimal disorder, predominant RMDD and GAD, "highly comorbid," and PD/AG and TD. The nature and number of disorders represented suggests that the first classes are distributed ordinarily on a dimension of severity, and the fourth class is qualitatively distinct. Support for this structure is based on the number of disorders, age at onset of OCD, neuroticism, and extraversion. CONCLUSIONS: In this OCD enriched sample, LCA identified four classes of disorder. These classes appear to conform to two subgroups that may prove useful in investigating the etiology of OCD. PMID- 12742680 TI - Neurocognitive functioning in subjects with eating disorders: the influence of neuroactive steroids. AB - BACKGROUND: Neuropsychological studies in subjects with eating disorders (EDs) have reported conflicting findings, which might be accounted for by several confounding variables, including neuroendocrine changes. METHODS: General abilities, executive functions, attention, and noneffortful learning were assessed in 45 patients with EDs and 45 healthy comparison subjects (HCS). Plasma levels of 17beta-estradiol, cortisol, allopregnanolone, dehydroepiandrosterone and its sulfate metabolite (DHEA and DHEAS) were evaluated in a subsample of patients and HCS. The influence of clinical, demographic, and neuroendocrine variables on neurocognitive performance was explored. RESULTS: Patients were slower than HCS on noneffortful learning and more accurate on a spatial executive task. DHEA and DHEAS were increased and positively correlated with accuracy on the executive task, while cortisol positively correlated with speed of noneffortful learning. CONCLUSIONS: A subtle impairment of noneffortful learning is the only neuropsychological deficit in patients with EDs. Changes in neuroactive steroids influence neurocognitive performance. PMID- 12742682 TI - Impact factors of forensic science and toxicology journals: what do the numbers really mean? AB - This article presents review and opinion about the use and abuse of journal impact factors for judging the importance and prestige of scientific journals in the field of forensic science and toxicology. The application of impact factors for evaluating the published work of individual scientists is also discussed. The impact factor of a particular journal is calculated by dividing the number of current year citations to a journal's articles that were published in the previous 2 years by the total number of citable items (articles and reviews) published in the same 2-year period. Journal impact factors differ from discipline to discipline and range from 0 for a journal whose articles are not cited in the previous 2 years to 46 for a journal where the average recent article is cited 46 times per year. The impact factor reflects the citation rate of the average article in a journal and not a specific article. Many parameters influence the citation rate of a particular journal's articles and, therefore, its impact factor. These include the visibility and size of the circulation of the journal including availability of electronic formats and options for on-line search and retrieval. Other things to consider are editorial standards especially rapid and effective peer-reviewing and a short time lag between acceptance and appearance in print. The number of self-citations and citation density (the ratio of references to articles) and also the inclusion of many review articles containing hundreds of references to recently published articles will boost the impact factor. Judging the importance of a scientist's work based on the average or median impact factor of the journals used to publish articles is not recommended. Instead an article-by-article citation count should be done, but this involves much more time and effort. Moreover, some weighting factor is necessary to allow for the number of co-authors on each article and the relative positioning of the individual names should also be considered. Authors should submit their research results and manuscripts to journals that are easily available and are read by their peers (the most interested audience) and pay less attention to journal impact factors. To assess the true usefulness of a person's contributions to forensic science and toxicology one needs to look beyond impact factor and citation counts. For example, one might consider whether the articles contained new ideas or innovations that proved useful in routine forensic casework or are widely relied upon in courts of law as proof source. PMID- 12742683 TI - Gamma hydroxybutyric acid (GHB) concentrations in humans and factors affecting endogenous production. AB - The endogenous nature of the drug of abuse gamma hydroxybutyric acid (GHB) has caused various interpretative problems for toxicologists. In order to obtain data for the presence of endogenous GHB in humans and to investigate any factors that may affect this, a volunteer study was undertaken. The GHB concentrations in 119 urine specimens from GHB-free subjects and 25 urine specimens submitted for toxicological analysis showed maximal urinary GHB concentrations of 3mg/l. Analysis of 15 plasma specimens submitted for toxicological analysis detected no measurable GHB (less than 2.5mg/l). Studies in a male and female volunteer in which different dietary food groups were ingested at weekly intervals, showed significant creatinine-independent intra-individual fluctuation with overall urine GHB concentrations between 0 and 2.55, and 0 and 2.74mg/l, respectively. Urinary concentrations did not appear to be affected by the particular dietary groups studied. The concentrations measured by gas chromatography with flame ionisation detection (GC-FID) and gas chromatography with mass spectrometry (GC MS) lend further support to the proposed urinary and plasma interpretative cut offs of 10 and 4mg/l, respectively, where below this it is not possible to determine whether any GHB detected is endogenous or exogenous in nature. PMID- 12742684 TI - Forensic cases involving the use of GHB in The Netherlands. AB - In this study, forensic cases involving the use of Gamma Hydroxy Butyric acid (GHB) from the second half of 1999 through the second half of 2001 in The Netherlands (blood >5mg/l and urine >10mg/l) are described. GHB was analysed by GC-MS after lactone formation and using GHB-d6 as internal standard. The results are divided into three groups: cases of chemical submission, cases of driving under the influence and cases of unknown causes of death.GHB was found in six cases of possible chemical submission. In these cases, relatively low concentrations of GHB were found. The results show that in cases of chemical submission, urine should be analyzed, because GHB is present longer in urine than in blood. The police should collect the samples in containers that do not contain citrate as anticoagulant. Especially at low levels of GHB, the formation of GHB in these tubes hampers an interpretation of the results.GHB was found in 13 cases of driving under the influence. In contrast to the cases of chemical submission, high concentrations of GHB were found, corresponding with observations of extreme sleepiness or temporary loss of consciousness.GHB was found in 16 cases of unexplained death: the measured range of GHB concentrations in blood might correspond to effects such as drowsiness, but not to serious toxicity of GHB. In 4 of these 16 cases, the role of GHB could be excluded. In the remaining cases, the role of GHB remains unclear; more research into "background" concentrations of GHB in post-mortem material is required. The incidence of the use of GHB in The Netherlands cannot be derived from these toxicological data. As GHB is not routinely found during systematical toxicological analyses, these data may seriously underestimate the use of GHB. Therefore, information from the police to the forensic institute is essential. PMID- 12742685 TI - High prevalence of 6-acetylmorphine in morphine-positive oral fluid specimens. AB - Identification of 6-acetylmorphine, a specific metabolite of heroin, is considered to be definitive evidence of heroin use. Although 6-acetylmorphine has been identified in oral fluid following controlled heroin administration, no prevalence data is available for oral fluid specimens collected in the workplace. We evaluated the prevalence of positive test results for 6-acetylmorphine in 77,218 oral fluid specimens collected over a 10-month period (January-October 2001) from private workplace testing programs. Specimens were analyzed by Intercept immunoassay (cutoff concentration=30 ng/ml) and confirmed by GC-MS-MS (cutoff concentrations=30 ng/ml for morphine and codeine, and 3 ng/ml for 6 acetylmorphine). Only morphine-positive oral fluid specimens were tested by GC-MS MS for 6-acetylmorphine. A total of 48 confirmed positive morphine results were identified. An additional 107 specimens were confirmed for codeine only. Of the 48 morphine-positive specimens, 32 (66.7%) specimens were positive for 6 acetylmorphine. Mean concentrations (+/-S.E.M.) of morphine, 6-acetylmorphine and codeine in the 32 specimens were 755+/-201, 416+/-168 and 196+/-36 ng/ml, respectively. Concentrations of 6-acetylmorphine in oral fluid ranged from 3 to 4095 ng/ml. The mean ratio (+/-S.E.M.) of 6-acetylmorphine/morphine was 0.33+/ 0.06. It is suggested that, based on controlled dose studies of heroin administration, ratios >1 of 6-acetylmorphine/morphine in oral fluid are consistent with heroin use within the last hour before specimen collection. The confirmation of 6-acetylmorphine in 66.7% of morphine-positive oral fluid specimens indicates that oral fluid testing for opioids may offer advantages over urine in workplace drug testing programs and in testing drugged drivers for recent heroin use. PMID- 12742686 TI - Urinary excretion profiles of 11-nor-9-carboxy-Delta9-tetrahydrocannabinol: a Delta9-THC-COOH to creatinine ratio study #2. AB - Subjects with a history of chronic marijuana use were screened for cannabinoids in urine specimens with the EMIT((R)) II Plus cannabinoids assay with a cut-off value of 50 ng/ml. All presumptively positive specimens were submitted for confirmatory analysis for the major urinary cannabinoid metabolite (Delta(9)-THC COOH) by GC-MS with a cut-off value of 15 ng/ml. Creatinine was analyzed in each specimen as an index of dilution. Huestis and Cone [J. Anal. Toxicol. 22 (1998) 445] reported that serial monitoring of Delta(9)-THC-COOH to creatinine ratios in paired urine specimens collected at least 24h apart could differentiate new drug use from residual Delta(9)-THC-COOH excretion. The best accuracy (85.4%) for predicting new marijuana use was a Delta(9)-THC-COOH/creatinine ratio > or =0.5 (dividing the Delta(9)-THC-COOH to creatinine ratio of specimen 2 by the specimen 1 ratio). In a previous study in this laboratory [J. Anal. Toxicol. 23 (1999) 531], urine specimens were collected from chronic marijuana users at least 24h apart and dilute urine specimens (creatinine values <2.2 micromol/l) were excluded from the data analysis. The objective of the present study was to determine whether creatinine corrected urine specimens positive for cannabinoids could differentiate new marijuana use from the excretion of residual Delta(9)-THC COOH in chronic users of marijuana based on the Huestis 0.5 ratio. Urine specimens (N=946) were collected from 37 individuals with at least 48h between collections. All urine specimens were included in the data review irrespective of creatinine concentration. The mean urinary Delta(9)-THC-COOH concentration was 302.4 ng/ml, mean Delta(9)-THC-COOH/creatinine ratio (ng/ml Delta(9)-THC COOH/(mmol/l) creatinine) was 29.3 and the Huestis ratio calculation indicated new drug use in 83% of all sequentially paired urine specimens. The data were sub divided into three groups (A-C) based on the mean Delta(9)-THC-COOH/creatinine values. Interindividual Delta(9)-THC-COOH/creatinine mean values ranged from 2.2 to 13.8 in group A (264 specimens, N=15 subjects) where 80.7% of paired specimens indicated new drug use. In group B, mean Delta(9)-THC-COOH/creatinine values ranged from 15.3 to 37.8 in 444 specimens (N=14 subjects) and 83.3% of paired specimens indicated new drug use. In group C, individual mean Delta(9)-THC COOH/creatinine values were >40.1 (41.3-132.5) in 238 urine specimens (N=8 subjects) and 85.3% of paired urine specimens indicated new marijuana use. Correcting Delta(9)-THC-COOH excretion for urinary dilution and comparing Delta(9)-THC-COOH/creatinine concentration ratios of sequentially paired specimens (collected at least 48h apart) provided an objective indicator of new marijuana use in this population. PMID- 12742687 TI - Automated headspace solid-phase dynamic extraction for the determination of cannabinoids in hair samples. AB - This article describes a fully automated procedure for detecting cannabinoids in human hair samples. The procedure uses alkaline hydrolysis and headspace solid phase dynamic extraction (HS-SPDE), followed by on-coating derivatization and gas chromatography-mass spectrometry (GC-MS). SPDE is a further development of solid phase microextraction (SPME), based on an inside needle capillary absorption trap. It uses a hollow needle with an internal coating of polydimethylsiloxane as extraction and pre-concentration medium. Ten mg of hair were washed with deionised water, petroleum ether and dichloromethane. After adding deuterated internal standards, the sample was hydrolyzed with sodium hydroxide and directly submitted to HS-SPDE. After absorption of analytes for an on-coating derivatization procedure, the SPDE-needle was directly placed into the headspace of a second vial containing N-methyl-N-trimethylsilyl-trifluoroacetamide before GC-MS analysis. The limit of detection was 0.14 ng/mg for Delta(9) tetrahydrocannabinol, 0.09 ng/mg for cannabidiol, and 0.12ng/mg for cannabinol. Absolute recoveries were in the range of 0.6 to 8.4%. Linearity was verified over a range from 0.2 to 20 ng/mg, with coefficients of correlation between 0.998 and 0.999. Intra- and inter-day precision were determined at two different concentrations and resulted in ranges between 2.3 and 6.0% (intra-day) and 3.3 and 7.6% (inter-day). Compared with conventional methods of hair analysis, this automated HS-SPDE-GC-MS procedure is substantially faster. It is easy to perform without using solvents and with minimal sample quantities, and it yields the same sensitivity and reproducibility. Compared to SPME, we found a higher extraction rate, coupled with a faster automated operation and greater stability of the device. PMID- 12742688 TI - Determination of cathinone, cathine and norephedrine in hair of Yemenite khat chewers. AB - A sensitive and reproducible method for the quantitative determination of cathinone (CTN), norpseudoephedrine (NPE, cathine) and norephedrine (NE) from hair was developed. The compounds were extracted for 4 hours with phosphate buffer pH 2.0, followed by a standard solid phase extraction procedure on a mixed phase column, derivatization with heptafluorobutyric anhydride (HFBA) and GC-MS separation and quantification using D(3)-ephedrine (D(3)-E) and alpha aminoacetophenone (AAP) as the internal standards. The diastereomers NPE and NE were satisfactorily separated. In the validation, the limits of detection and of quantification were determined at 0.03-0.08 ng/mg and 0.10-0.24 ng/mg, respectively and the interday standard deviation was between 10 and 15%. The method was applied to hair samples of 24 Yemenite khat chewers. All three compounds were detected in 23 of these cases. The concentrations ranged from 0.57 to 23.9 ng/mg for NPE, 0.19-25.0 ng/mg for NE and 0.11-22.7ng/mg for CTN. A highly significant correlation was found between the self-reported data about the khat consumption habits of the volunteers (4-56h chewing per week) and the concentrations of norephedrine and norpseudoephedrine in hair. PMID- 12742689 TI - A study into the rate of incorporation of eight benzodiazepines into rat hair. AB - The incorporation of eight benzodiazepines (chlordiazepoxide, diazepam, estazolam, flunitrazepam, flurazepam, medazepam, oxazepam and triazolam) into rat hair was investigated by HPLC and GC-MS. Each of the benzodiazepines was injected daily into three Dark Agouti (DA) rats for 10 days at 10mg/kg. The back hair of the rats was removed by shaving prior to the first injection and again on the 28th day after the initial administration. To investigate optimum extraction conditions, 10mg aliquots of rat hair incorporated with diazepam, flurazepam or medazepam were extracted by seven different methods (Proteinase K, methanol ammonia, methanol-trifluoroacetic acid, Soerensens buffer, 1M NaOH, beta glucuronidase/arylsulfatase, Biopurase). The method found to yield the highest recoveries, for all three drugs, was the acidic methanol extraction. Using this extraction procedure, the incorporation rates (ICR: the ratio of the hair concentration to the plasma AUC) of eight benzodiazepines into rat hair were investigated. The ICRs ranged from 0.002 (flunitrazepam) to 0.049 (flurazepam). The major metabolites of flurazepam were investigated in rat hair. The mean hair concentrations of desalkylflurazepam and 2-hydroxyethylflurazepam were 3.31 and 0.05 ng/mg, respectively, which are 24 and 0.36% of the parent compound in hair. PMID- 12742690 TI - Hair analysis of opiates in mothers and newborns for evaluating opiate exposure during pregnancy. AB - The increasing interest in toxicological hair analysis as a marker of human exposure to xenobiotics such as illicit substances or therapeutic drugs, has been made feasible by the extension of mass spectrometry, a highly sensitive method of detection. A newborn exposed to drugs in utero can suffer from a varying degree of withdrawal syndrome, a few days after birth. If of opiate origin, the withdrawal syndrome can be treated with morphine, among other therapeutics, but it is not easy to diagnose because of atypical symptoms presented by neonates and especially when maternal drug addiction has not been revealed. To assess and measure toxicological factors linked with the appearance and the severity of this syndrome, maternal and neonatal matrices such as urine, meconium and hair were collected during a protocol approved by the ethical committee. Opiates in particular were measured with GC-MS and potential combined dependences (cannabis, cocaine, amphetamine, LSD and benzodiazepines) and/or substitutive therapeutics (methadone or buprenorphine) were also assessed in 17 mother/neonate couples. Gestational opiate exposure profiles were drawn up and linked with the observed withdrawal syndromes. A withdrawal syndrome seems to appear more frequently after foetal exposure to an association of opiates/substitutive molecules (8 out of 10 withdrawal syndromes observed in this study), although the impact of cocaine and benzodiazepines must also be taken into account. The results obtained in neonatal hair make it possible to affirm foetal drug exposure and are in accordance, for the majority, with the appearance of a neonatal withdrawal syndrome (NWS). Neonatal hair analysis could contribute to assess in utero exposure to opiates, particularly when results in urine and meconium are negative or when these matrices are not available. PMID- 12742691 TI - Comparison of daily urine, sweat, and skin swabs among cocaine users. AB - This study (1) compares urine, skin swabs, and PharmChek sweat patches for monitoring drug use; (2) measures possible environmental contamination in recent cocaine (COC) users; and (3) evaluates various immunoassays (IA) for screening COC in diverse matrices. Unique aspects include daily urine monitoring of 10 participants for 4 weeks, multiple monitoring methods, analysis for all specimens by IA and gas chromatography (GC)/mass spectrometry (MS), and the potential for continued illicit drug use by participants. Urine served as the "gold standard" specimen for determining drug use. Only cocaine and related substances were detected. Trace amounts of drugs were found on the skin (<50 ng per swab) of urine-negative participants' hands or forehead. In contrast, larger quantities of COC were found on the skin of individuals with BE-positive urines or individuals living with drug users (up to 20 microg per swab). Patch COC amounts among the three regular users (250-9000, 0-240, 160-22,000 ng per patch) exceeded BE (50 950, none, 30-2200 ng per patch). Pre-swabs, valuable for interpreting the source or time frame of positive patch results, contained substantial COC (38-1160, 0 152, 34-762 ng per swab) prior to patch application; therefore, patch results may represent current use, prior use, contamination, or a combination. In three individuals with no indication of cocaine use, false positives (defined as sweat patch positive when urine specimens were <300ng BE/ml) occurred at a 7% rate. Proposed cut-off concentrations of 75 ng cocaine per patch and 300 ng BE/ml urine curtail the incidence of false positives in this limited population. Three immunoassays were compared to screen specimens for cocaine: a modified, manual Microgenics CEDIA; a Cozart ELISA; and an OraSure ELISA. CEDIA's limit of detection (LOD) was 81ng/ml, compared with LODs of 4 ng/ml for the Cozart ELISA and 1.5 ng/ml for the OraSure ELISA. Cozart correlated with OraSure results for COC concentrations <2000 ng per swab (n=117), r(2)=0.79. PMID- 12742693 TI - Ethyl glucuronide concentrations in two successive urinary voids from drinking drivers: relationship to creatinine content and blood and urine ethanol concentrations. AB - The concentrations of alcohol in blood (BAC) and two successive urine voids (UAC) from 100 drunk drivers were compared with the concentration of ethyl glucuronide (EtG), a minor metabolite of ethanol in urine, and the urinary creatinine content as an indicator of dilution. The subjects consisted of 87 men with mean age 42.2+/-14.2 years (+/-standard deviation, S.D.) and 13 women with mean age 42.5+/ 14.4 years. Ethanol was measured in blood and urine by headspace gas chromatography (GC) and EtG was determined in urine by liquid chromatography-mass spectrometry (LC-MS). The mean UAC was 2.53+/-1.15g/l for first void compared with 2.35+/-1.17g/l for second void, decreasing by 0.18+/-0.24g/l on average (P<0.001 in paired t-test). The ratios of UAC/BAC were 1.35+/-0.25 for first void and 1.20+/-0.16 for second void and the difference of 0.15+/-0.27 was statistically significant (P<0.001). The UAC/BAC ratio was not correlated with creatinine content of the urine specimens, whereas the concentration of urinary EtG was positively correlated with creatinine (r=0.64 for first void and r=0.62 for second void). The UAC was not correlated with urinary EtG directly (r=-0.03 for first void and r=0.08 for second void) but after adjusting for the relative dilution of the specimens (EtG/creatinine ratio) statistically significant positive correlations were obtained (r=0.58 for first void and r=0.57 for second void). The dilution of the urine, as reflected in creatinine content, is important to consider when EtG measurements are interpreted. The excretion of EtG in urine, like glucuronide conjugates of other drugs, is influenced by diuresis. EtG represents a sensitive and specific marker of acute alcohol ingestion with applications in clinical and forensic medicine. PMID- 12742692 TI - Comparison of the prevalence of alcohol, cannabis and other drugs between 900 injured drivers and 900 control subjects: results of a French collaborative study. AB - A collaborative case-control study was conducted in France in order to determine the prevalence of alcohol, cannabinoids, opiates, cocaine metabolites, amphetamines and therapeutic psychoactive drugs in blood samples from drivers injured in road accidents and to compare these values with those of a control population. Recruitment was performed in emergency departments of six university or general hospitals and comprised 900 drivers involved in a non-fatal accident and 900 patients (controls) who attended the same emergency units for a non traumatic reason. Drivers and controls were matched by sex and age. Alcohol was determined by flame ionization-gas chromatography, drugs of abuse (DOA) by gas chromatography-mass spectrometry with the same analytical procedures in the six laboratories, and medicines by high performance liquid chromatography with diode array detection. Blood alcohol concentration exceeding 0.5 g/l (i.e. the legal French threshold) was found in 26% of drivers and 9% of controls. In the 18-27 years age range, alcohol was the only toxic found in blood samples of 17% drivers and 5% controls, leading to an odds-ratio (OR) of 3.8. A significant relationship was found between alcohol blood concentrations and OR values. All age groups confounded, the main active substance of cannabis, Delta(9) tetrahydrocannabinol (THC), was found in 10% of drivers and 5% of controls. In the less than 27 years old, THC (>1 ng/ml) was detected alone in the blood of 15.3% drivers and of 6.7% controls, giving OR=2.5, whereas there was no link between THC blood concentrations and OR value. THC was found alone in 60% of cases and associated with alcohol in 32%, with OR=4.6 between drivers and controls for this association. The difference in morphine prevalence between drivers (2.7%) and controls (0.03%) was highly significant (P<0.001), with OR=8.2. The number of positive cases for amphetamines and cocaine metabolites was too low for reaching any interpretation. The most frequently observed psychoactive therapeutic drugs were by far benzodiazepines, that were found alone in 9.4% of drivers and 5.8% of controls, which led to OR=1.7 (P<0.01). This study demonstrates a higher prevalence of opiates, alcohol, cannabinoids and the combination of these last two compounds in blood samples from drivers involved in road accidents than in those from controls, which suggests a causal role for these compounds in road crashes. PMID- 12742694 TI - A fatal forensic intoxication with fenarimol: analysis by HPLC/DAD/MSD. AB - Fenarimol (Rubigan) is a pyrimidine ergosterol biosynthesis inhibitor used as a systemic fungicide. The authors present a fatal fenarimol intoxication case analysed in the Forensic Toxicology Service of the National Institute of Legal Medicine. The results were used to compare two different HPLC techniques, regarding selectivity and sensitivity: an HPLC system with a diode array detector (DAD) and an HPLC system with a DAD and a mass spectrometry detector (MSD) with an electrospray interface. All biological samples were submitted to a solid-phase extraction procedure. The detection and quantification limits of fenarimol, linearity, precision and accuracy were evaluated. The fenarimol concentration levels determined were of 89.0 mg/ml in gastric contents, 1.9 mg/g in liver and 0.4 mg/g in kidney. Blood was not available at autopsy. No published data related to fenarimol self-poisoning were found, so it was not possible to interpret the results obtained by comparison with toxic/lethal levels. PMID- 12742695 TI - LC-MS/MS analysis of pholedrine in a fatal intoxication case. AB - Pholedrine (4'-hydroxymethamphetamine) is a cardiovascular agent exerting hypertensive and adrenergic effects. High doses may cause a drop in the peripheral circulation blood flow and increase blood pressure, heart rate and body temperature up to a state of central respiratory paralysis. A 15-year-old girl who suffered from heavy agitation and hallucinations was admitted to the intensive care unit in a comatose state. The clinical findings included a maximum heart rate of 170 bpm and a body temperature of 43.8 degrees C. Resuscitation measures were in vain and abandoned after approximately 2h. A toxicological emergency analysis using GC/MS revealed a considerable amount of pholedrine in blood and urine. A method for determining pholedrine in human body fluids utilizing high-performance liquid chromatography (HPLC)/tandem mass spectrometry (LC-MS/MS) with a turbo ion-spray source was developed, using D11-methamphetamine and D5-methylenedioxymethamphetamine as internal standards. Samples were prepared by SPE extraction using SPEC-C18AR/MP3((R)) columns, which yielded the best extraction recovery (67%). Chromatographic separation was achieved at pH 5 on an RP-18 stationary phase applying gradient elution from 50 to 70% of B (methanol/acetonitrile 3/1 (v/v), 0.02% acetic acid) in A (5mM ammonium acetate/acetonitrile 95/5 (v/v), 0.02% acetic acid). Supra-pure acetic acid was added to the post-column effluent with a flow rate of 0.2 microl/min to optimize ionization. Detection was carried out in the positive ionization, multiple reaction monitoring (MRM) mode. The chromatograms showed no interference from other substances. The limit of detection (LOD, S/N=3) of pholedrine was 0.8 ng/ml and its lower limit of quantification (LLOQ, S/N=10) 3ng/ml. The calibration curve was linear (r=0.999) in the range 1-100 ng/ml. Samples with higher concentrations were diluted to suit the working range. The intra-day R.S.D. between 5 and 80 ng/ml were 3.8-8.7% and the inter-day R.S.D. between 5 and 100 ng/ml were 6.7-10.7%. The pholedrine concentrations in blood and urine collected when the girl was still alive were 16.1 microg/ml (R.S.D. 10.5%) and 1120 microg/ml (R.S.D. 8%), respectively. In post-mortem samples, they were 23.0 microg/ml (R.S.D. 5.1%) in heart blood and 27.3 microg/g (R.S.D. 6.6%) in the liver. PMID- 12742697 TI - A multiple drug fatality involving MK-801 (dizocilpine), a mimic of phencyclidine. AB - MK-801 (dizocilpine) is a non-competitive antagonist at the N-methyl-D-aspartate (NMDA) family of glutamate receptors in the central nervous system. It is an anticonvulsant and also shares several pharmacological properties with phencyclidine and ketamine. It is not observed routinely as a substance of abuse. The deceased, a 45-year-old white male, obtained MK-801 surreptitiously in an attempt to treat a self-diagnosed depression. He was discovered the next morning, unresponsive on the bathroom floor. An empty bottle, labeled to contain 25mg of MK-801, was found near the body. The autopsy was performed at the Joseph A Jachimczyk Forensic Center, Houston, TX. Body weight at autopsy was 88kg. Lungs were edematous and congested (right: 775g; left 700g). The heart had proportionate chambers and was otherwise unremarkable. The kidneys (right: 220g; left 225g) were smooth surfaced. The brain (1550g) was congested and without trauma. Microscopic evaluation of the heart, kidneys and lungs showed normal histology and confirmed pulmonary congestion and edema. Samples of heart blood, liver, bile, vitreous humor, stomach contents and urine were collected at autopsy. There were 550ml of stomach contents. Drugs in blood were screened by EMIT II Plus immunoassay procedures and by gas chromatography/mass spectrometry (GC/MS) of an organic solvent extract of basified blood. Alcohol was determined by gas chromatography with headspace injection. MK-801, benzodiazepines and alcohol were detected in blood. Amounts of MK-801 present in blood, bile, liver, vitreous humor and urine were 0.15, 0.29, 0.92, less than 0.1 and 0.36 mg/l (kg), respectively. The cause of death was benzodiazepine, dizocilpine and ethanol toxicity and the manner accidental. PMID- 12742696 TI - An autopsy case of combined drug intoxication involving verapamil, metoprolol and digoxin. AB - We present here a fatal poisoning case involving verapamil, metoprolol and digoxin. A 39-year-old male was found dead in his room, and a lot of empty packets of prescribed drugs were found near the corpse. The blood concentrations of verapamil, metoprolol and digoxin were 9.2 microg/ml, 3.6 microg/ml and 3.2 ng/ml, respectively. The cause of death was given as cardiac failure, hypotension and bradycardia due to a mixed drug overdose of verapamil, metoprolol and digoxin, based on the results of the autopsy and toxicological examination. We speculate that the toxicity of verapamil is potentiated by drug interaction with metoprolol and digoxin. PMID- 12742698 TI - Methadone substitution: medicolegal problems in Germany. AB - In Germany, the substitution of methadone for heroin abusers has arisen recently and has resulted in various medicolegal problems. Normally, these concern methadone-associated deaths, or the prescription of methadone resulting in criminal prosecutions concerning physicians. Not to be forgotten is the problem of driving while taking methadone. In the years 1997-2001, we detected methadone in the blood of 398 cases that were analysed by the Institute of Legal Medicine, Bonn. Methadone was the only drug in only 18 cases. In most of the cases, up to five additional drugs were also being taken: benzodiazepines (61%), ethanol (40%), morphine (39%), cannabinoids (35%), cocaine (28%), anti-depressants (3%), and amphetamines (2%). PMID- 12742699 TI - Cases of fatal poisoning in post-mortem examinations at the Institute of Forensic Medicine in Greifswald--analysis of five decades of post-mortems. AB - Apparently, fatal poisoning as cause of death are still rarely found in unnatural deaths investigated in the institutes of forensic medicine. In the Institute of Forensic Medicine at the University of Greifswald, 10-15% of the post-mortem autopsies displayed an intoxication during the last several decades with a possible decreasing tendency. Thirteen thousand eight hundred and nineteen autopsies were carried out in our institute-situated in a low populated rural area-during the last 50 years with the confirmed death cause intoxication in 1,589 times. In this study, especially the intoxication causes and the substance classes of the poisonous agents have been investigated. In addition, we analyzed the frequency of intoxications as well as sex and age of the deceased. Surprisingly, CO-intoxications were found most frequently with an incidence of 49% followed by alcohol intoxications with 21%. The latter was not unexpected taking into account the habits of the local population. Medical drugs and narcotics take only the third place, although the abuse of modern narcotics drugs is already visible even in the far east of Germany. The spectrum of substances which are abused, taken accidentally or deliberately is continuously changing, reflecting scientific progress in the pharmaceutical industry as well as fashion tendencies. Therapeutic use is almost always followed by abuse. Our results confirm prior experience concentrating mostly on other poisons like heavy metals or herbicides, etc. In addition, we could demonstrate the influence of political conditions on use and distribution of illegal drugs in Germany. Our study clearly demonstrates that insufficient equipment or analytical methods are no longer the reason for any problems uncovering lethal intoxications. They are rather due to insufficient investigations of the corpses (without considering the possibility of an intoxication as differential diagnosis) and to frequent mistakes of the prosecutor's office in death without signs of physical violence. These facts may explain the above mentioned decreasing tendency of intoxications, but they also clarify that this tendency probably does not correspond to reality. PMID- 12742700 TI - Identification and quantitation of xenobiotics by 1H NMR spectroscopy in poisoning cases. AB - In order to analyse a wide range of xenobiotics and their metabolites present in biological fluids, NMR spectroscopy can be used. A large variety of xenobiotics (therapeutic agents, pesticides, solvents, alcohols) can be characterized and quantitated directly, without sample preparation. NMR investigations were applied to acute poisoning cases, involving drugs such as salicylates and valproic acid (VPA). In a salicylate poisoning case, the three major metabolites of acetylsalicylic acid have been detected in crude urine, and rapid identification of lysine revealed the origin of the intoxication, namely lysine acetylsalicylate (Aspegic). Valproic acid as its glucuronide was identified in urine samples from two poisoned patients. 1H NMR was also used to identify and quantitate paraquat (Gramoxone) in urine owing to its two aromatic signals at 8.49 and 9.02 ppm, in two acutely poisoned patients (183 and 93 mg/l). An intentional poisoning case with tetrahydrofuran (THF) was also investigated. Serum and urine samples were collected. THF was characterized by its resonances at 1.90 and 3.76 ppm, and quantified at 813 and 850 mg/l in the two biological fluids, respectively. Moreover, two other compounds were detected: lactate and gamma-hydroxybutyric acid (GHB). 1H NMR spectroscopic analysis of serum samples from three poisoned patients revealed methanol in one case and ethylene glycol in the two others. Moreover, in the same spectrum, the corresponding metabolites formate and glycolate were found. Compared with the reference chromatographic or spectrophotometric methods, requiring time-consuming extraction and/or derivatization steps, NMR spectroscopy allows the determination of many exogenous and endogenous compounds, without any pre-selection of the analytes. PMID- 12742701 TI - Human serum paraoxonase (PON1) activity in acute organophosphorous insecticide poisoning. AB - Human serum paraoxonase (PON1) and perhaps other mammalian paraoxonases catalyzes the hydrolysis of certain organophosphorus (OP) insecticides and nerve gases and so may alter significantly an individual's susceptibility to the toxicity of these chemicals. Serum PON1 exhibits a substrate dependent polymorphism and this polymorphism shows great interethnic variability. This study focused on the investigation of PON1, arylesterase and cholinesterase activities in 28 acute OP insecticide poisoning cases. Insecticide analysis were performed by GC-NPD and activities of enzymes were measured by using spectrophotometer. The activity levels for salt stimulated PON1, basal PON1 and arylesterase were found as 78.83 (35.39-186.13), 39.97 (2.49-80.43) micromol/min/l and 126.26 (36.34-288.24) mmol/min/l respectively. On the other hand the activity levels for butyrylcholinesterase (BTC) and acetylcholinesterase (AchE) were found as 797.23 (106.3-3823)U/l and 4.65 (0.21-30.29)U/ml. There was a correlation between percent stimulation of PON1 and BTC activities (r=0.446, P<0.05), but this correlation was lower than in cases who exposed to OP insecticides chronically. As a conclusion, in chronic and acute OP exposure, both PON1 level and phenotype must be taken into consideration. PMID- 12742702 TI - Dermal absorption of kerosene components in rats and the influence of its amount and area of exposure. AB - The influences of amount and area of dermal exposure to kerosene upon the levels of kerosene components in biological samples were examined in vivo and in vitro. Thirty-two rats were randomly divided into four groups and exposed to kerosene through the abdominal skin for 2h. The amounts (soaked in cotton) and area of kerosene exposed were 1 ml/4 cm(2) in Group I, 4 ml/4 cm(2) in Group II, 4 ml/16 cm(2) in Group III and 16 ml/64 cm(2) in Group IV. Before, then 5, 10, 20, 30, 45, 60, 90 and 120 min after exposure, 0.5 ml of blood was collected. Solid tissue samples, including the exposed skin area, were harvested at 120 min. Kerosene components were analyzed by gas chromatography/mass spectrometry. Trimethylbenzens (TMBs) that are easily absorbed kerosene components, appeared at 5-20 min. The time course changes in TMB levels in blood were significantly different between Groups I and II or Groups I and III, and almost identical between Groups II and III. Similar trends were observed in tissue samples at 120 min. High concentrations of aliphatic hydrocarbons (AHCs) were detected in the exposed skin and the AHC levels were dependent on the amount of kerosene exposed per unit area. These results suggest that (1) dermal absorption of kerosene occurs soon after dermal exposure started, (2) absorption of TMBs is influenced by the total amount of kerosene rather than area of exposure, and (3) AHCs remaining in the skin at significant levels are influenced by the amount of kerosene per unit area exposed. PMID- 12742704 TI - Post-mortem analysis of formic acid disposition in acute methanol intoxication. AB - Fifteen cases of fatal massive methanol intoxication have been investigated. Victims received either no treatment or ethanol therapeutic treatment. Methanol poisoning cases were classified in three groups according to survival time: more than 3 days (group 1), up to 3 days (group 2) and few hours (group 3). Body distribution of methanol and formic acid, as the main metabolite, was analyzed in blood and in different organs (brain, kidney, lung and liver). Relationships between formic acid concentration in the different tissues, survival time and type of treatment applied to victims were studied. Formic acid in blood and tissues was analyzed by head space gas chromatography (head space-GC) with FID detector, previous transformation in methyl formate, essentially as described by Abolin. Formic acid concentration was between 0.03 and 1.10g/l in the samples under study. A good correlation between blood and brain, but poor between blood and the remaining tissues was found. Obtained data suggested that the use of blood and brain could help to improve the analysis of formic acid intoxication. The best correlation among organs was found between lung and kidney for all groups (r(2)=0.91, 0.84 and 0.87, corresponding to groups 1, 2 and 3, respectively). Lethality index was defined as LI = (concentration of formic acid in blood in (g/l)/0.5) x 100, taking into account that 0.5g/l is the concentration reported by Mahieu in severe methanol poisoning. LI parameter was used to estimate formic acid incidence on the lethality of methanol poisoning cases. LI showed a good correlation with total formic acid concentration of the different tissues analyzed (r(2)=0.80). Furthermore, LI allowed us to discriminate between individuals that received therapeutic treatment and survived different periods. LI>100 indicated a severe intoxication and short survival time if the victim was assisted with ethanol therapy and hemodialysis was not applied. With regard to victims who received no therapeutic treatment and died in few hours, LI was in the range 40-100. LI was below 40 for individuals that survived more than 3 days and hemodialysis was not performed. Results showed the importance of performing formic acid analysis to diagnose severe methanol intoxication in post-mortem cases. PMID- 12742703 TI - Hippuric acid and methyl hippuric acid in rat hair: possible monitoring of xylene and toluene exposure. AB - Thinner is mainly composed of toluene and xylenes, and we studied the incorporation of the main metabolites of toluene and xylenes, hippuric acid (HA) and o-, m-, and p-methyl hippuric acids (o-, m-, p-MHA), in dark agouti rats' hair. Rat black hair was shaved before any exposure with an electric shaver designed for animals. Studies were performed in vivo with exposures of 30 min per day at three different concentrations (100, 300, and 1000 ppm) of toluene and o-, m-, and p-xylene for a total of 10 times over 2 weeks. Newly grown hair was tweezed out from the root with tweezers at seventh of the last exposure. Hair samples were then washed, extracted, derivatized, and analyzed by gas chromatography-mass spectrometry (GC-MS). HA and o-, m-, and p-MHA were not detected (ND) in the unexposed rat hair. After exposure, the metabolite concentration in the hair changed depending on the exposure concentration. Mean concentrations ranged from ND to 7.6 ng/mg, from ND to 13.8 ng/mg, from ND to 10.1 ng/mg, and from ND to 9.2 ng/ml hair for HA, o-, m-, and p-MHA, respectively. These results indicate that the metabolites concentrations in hair are effective indices of thinner exposure. PMID- 12742705 TI - Anise oil as para-methoxyamphetamine (PMA) precursor. AB - These days, MDMA is one of the most popular drugs of abuse. Due to its illegality, MDMA and its chemical precursors are watched by governmental organizations in many countries. To avoid conflicts with legal instances, underground chemists have tried to market several new unregulated amphetamine analogues, such as 4-MTA. Para-methoxyamphetamine (PMA), on the other hand, is regulated by law but its precursors are easily obtained since they are cheap and unwatched. This article presents such a case, namely the large scale synthesis of PMA using anethole, a main constituent of anise oil, as precursor. Anethole has been converted to its phenyl acetone analogue via peracid oxidation, while PMA itself has been synthesized using this ketone as precursor in the Leuckart synthesis. The synthesis of PMA using anethole as starting product has been investigated applying GC/MS and GC-HSPME/MS techniques, hereby discovering new specific (4-methoxyphenol) and already identified synthesis impurities (4-methyl 5-(4-methoxyphenyl)pyrimidine, N-(beta-4-methoxyphenylisopropyl)-4-methoxybenzyl methyl ketimine, 1-(4-methoxyphenyl)-N-(2-(4-methoxyphenyl)-1-methylethyl-2 propanamine, 1-(4-methoxyphenyl)-N-methyl-N-(2-(4-methoxyphenyl)-1-methylethyl-2 propanamine, N-(beta-4-methoxyphenylisopropyl)-4-methoxybenzaldimine). The new impurity 4-methoxyphenol is specific for the application of a peracid oxidation method where anethole is used as precursor. PMID- 12742706 TI - Detection of thiopental and pentobarbital in head and pubic hair in a case of drug-facilitated sexual assault. AB - The quali-quantitative determination of two barbiturates, thiopental and its metabolite pentobarbital, in head and pubic hair samples of a woman who had been sexually assaulted during hospitalisation, is reported. Hair was analysed by means of solid-phase microextraction (SPME) and gas chromatography-multiple mass spectrometry (GC-MS-MS), in chemical ionisation conditions. Thiopental and pentobarbital were found in three proximal head hair segments (sample 1A: 0.30 and 0.40 ng/mg; sample 1B: 0.20 and 0.20 ng/mg; sample 3: 0.15 and 0.20 ng/mg) and pubic hair sample. Two distal head hair segments were negative for both barbiturates. Despite the lack of collection and toxicological analysis of blood or urine samples within the hospital setting, analytical findings from hair revealed the use of the anaesthetic agent thiopental to sedate the victim quickly and deeply and commit sexual assault. PMID- 12742707 TI - Proficiency test for the analysis of hair for drugs of abuse, organized by the Society of Hair Testing. AB - Eighteen laboratories interested in the analysis of human hair for drugs of abuse participated in a proficiency test (PT) organized by the Society of Hair Testing (SoHT) in 2001. Samples sent to the participants included one drug-free hair sample and two samples from drug users, sent in the form of short segments previously checked for homogeneity by three reference laboratories. Participants were requested to analyze the samples following the standard procedure used routinely in their laboratories.The compounds present in the samples included opiates, cocaine and metabolite, cannabinoids and amphetamines. All the laboratories analyzed opiates, cocaine and benzoylecgonine (BE); only 10 analyzed amphetamines, and 9 cannabinoids. Various methods were used to extract drugs from the hair-enzyme treatment, acidic, basic and methanol extractions. All the laboratories employed GC-MS, with the exception of two which used GC-MS/MS and LC MS/MS, respectively. Six laboratories performed initial screening tests by RIA, ELISA or EMIT. Results show that the laboratories performed well qualitatively, since they successfully identified all the analytes that they tested, with the exception of eight false results. However, the scatter of quantitative results was high. PMID- 12742708 TI - Rapid determination of carboxyhemoglobin in blood by Oximeter. AB - Different methods to determine carboxyhemoglobin (COHb) in blood are described in the literature. In our laboratory three methods to analyze COHb in post-mortem blood samples were compared: the spectrophotometric method of Maehly, a gas chromatographic method with a thermal conductivity detector (GC-TCD) and the Oximeter. Several COHb containing blood samples of deceased persons were analyzed. Results of all three methods were comparable for low concentrations (ca. 10% COHb) as well as for high concentrations (ca. 80% COHb) regardless of the viscosity of the blood samples. The advantages of the Oximeter when compared to Maehly's method and GC-TCD are extreme short time of analysis (<1min), very small blood volume required (<0.1ml) and easy handling. In our opinion application of the Oximeter is not limited to analyses of blood samples from living persons (e.g. in clinical toxicology); it can as well be used for the determination of COHb in post-mortem blood samples. Hence it is a useful and time saving tool in forensic toxicology. PMID- 12742710 TI - Natural infections of pigs with akabane virus. AB - Akabane (AKA) virus is considered a pathogen of herbivores in nature. However, we found that pig populations in fields were infected in Taiwan. An isolate (NT-14) of AKA virus was obtained from pigs. The NT-14 virus was able to infect pigs by the oronasal route. Subsequently, low levels of infectious virus particles were excreted into the oronasal discharge during the stage of viremia but they were not sufficient to infect new porcine hosts via contact transmission. The prevalence of serum neutralizing antibodies to AKA virus in pig populations was investigated, indicating that approximately 75% of pigs in Taiwan were seropositive. Sows and newborn piglets have the highest titers of neutralizing antibodies. Contrarily, fattening pigs aged at approximately 20 weeks old contained the lowest titers of specific antibodies. Our results suggest that pigs in natural situations are part of the AKA virus transmission cycle. PMID- 12742711 TI - Detection of a mammalian-like group A rotavirus in diarrhoeic chicken. AB - The present investigation describes detection of a mammalian-like electropherogroup A rotavirus in chicken with diarrhoea. This also records the first detection of a rotavirus in an avian species from India. During the investigation 75 diarrhoeic faecal samples collected from adult chicken were screened for the presence of group A rotavirus antigen by sandwich ELISA. All three samples positive for rotavirus antigen revealed 11 bands of RNA in polyacrylamide gel electrophoresis (PAGE). In contrast to avian group A rotavirus, segment 5 was found to migrate closer to 6 as is the case with mammalian group A rotaviruses. Segments 7, 8 and 9 were found to migrate as a tight triplet, which is characteristic of group A rotavirus. PMID- 12742713 TI - Cloning of the gene coding for Staphylococcus intermedius exfoliative toxin and its expression in Escherichia coli. AB - An exfoliative toxin (SIET)-producing strain (D-52) of Staphylococcus intermedius derived from canine pyoderma did not possess large plasmids. Therefore, the gene coding for SIET was considered to be located on the chromosomal DNA. The SIET gene was cloned from the chromosomal DNA of S. intermedius and was expressed in Escherichia coli. The nucleotide sequence of the SIET gene consists of a coding region of 990 bp specifying a polypeptide of 330 amino acid residues, which included a putative 42-residue signal sequence. PMID- 12742712 TI - Isolation of exfoliative toxin from Staphylococcus intermedius and its local toxicity in dogs. AB - A rounding effect was demonstrated in cultured cells inoculated with the culture filtrates (CFs) of 60 strains of Staphylococcus intermedius derived from dogs affected with pyoderma. Exfoliative toxin (ET)-like toxin (ETLT) was isolated from the CF of S. intermedius strain D-52, which exhibited strong rounding activity and then was purified by gel filtration on a Sephadex G-75 column, and by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The ETLT caused exfoliation in 1-day-old chickens, suckling Syrian hamsters, and dogs, but not in suckling mice. The ETLT was serologically different from exfoliative toxin A (ETA), exfoliative toxin B (ETB), exfoliative toxin C (ETC), S. hyicus exfoliative toxin A (SHETA), and SHETB, as shown by Western blot analysis. The molecular weight of the ETLT was estimated at 30 kDa by SDS-PAGE. In the present study, we propose the ETLT was a novel type of ET, S. intermedius exfoliative toxin (SIET). PMID- 12742714 TI - Antimicrobial susceptibilities of canine Clostridium difficile and Clostridium perfringens isolates to commonly utilized antimicrobial drugs. AB - Clostridium difficile and Clostridium perfringens are anaerobic, Gram-positive bacilli that are common causes of enteritis and enterotoxemias in both domestic animals and humans. Both organisms have been associated with acute and chronic large and small bowel diarrhea, and acute hemorrhagic diarrheal syndrome in the dog. The objective of this study was to determine the in vitro antimicrobial susceptibilities of canine C. difficile and C. perfringens isolates in an effort to optimize antimicrobial therapy for dogs with clostridial-associated diarrhea. The minimum inhibitory concentrations (MIC) of antibiotics recommended for treating C. difficile (metronidazole, vancomycin) and C. perfringens-associated diarrhea in the dog (ampicillin, erythromycin, metronidazole, tetracycline, tylosin) were determined for 70 canine fecal C. difficile isolates and 131 C. perfringens isolates. All C. difficile isolates tested had an MIC of or=256 microg/ml for both erythromycin and tylosin. A third C. perfringens isolate had an MIC of 32 microg/ml for metronidazole. Based on the results of this study, ampicillin, erythromycin, metronidazole, and tylosin appear to be effective antibiotics for the treatment of C. perfringens-associated diarrhea, although resistant strains do exist. However, because there is limited information regarding breakpoints for veterinary anaerobes, and because intestinal concentrations are not known, in vitro results should be interpreted with caution. PMID- 12742715 TI - Serotypes, phage types and virulence genes of shiga-producing Escherichia coli isolated from sheep in Spain. AB - PROBLEM ADDRESSED: Shiga toxin-producing Escherichia coli (STEC), have emerged as food poisoning pathogens which can cause severe diseases in humans. OBJECTIVE: The aim of this study was to determinate the serotypes and virulence genes of STEC strains isolated from sheep in Spain, with the purpose of determining whether sheep represent a potential source of STEC pathogenic for humans. METHODS AND APPROACH: Faecal swabs obtained from 697 healthy lambs on 35 flocks in Spain during the years 2000 and 2001 were examined for STEC using phenotypic (Vero cells) and genotypic (PCR) methods. RESULTS: STEC O157:H7 strains were isolated from seven (1%) animals in six flocks, whereas non-O157 STEC strains were isolated from 246 (35%) lambs in 33 flocks. A total of 253 ovine STEC strains were identified in this study. PCR showed that 110 (43%) strains carried stx(1) genes, 10 (4%) possessed stx(2) genes and 133 (53%) both stx(1) and stx(2). Enterohaemolysin (ehxA) and intimin (eae) virulence genes were detected in 120 (47%) and in 9 (4%) of the STEC strains. STEC strains belonged to 22 O serogroups and 44 O:H serotypes. However, 70% were of one of these six serogroups (O6, O91, O117, O128, O146, O166) and 71% belonged to only nine serotypes (O6:H10, O76:H19, O91:H-, O117:H-, O128:H-, O128:H2, O146:H21, O157:H7, O166:H28). A total of 10 new O:H serotypes not previously reported in STEC strains were found in this study. Seven strains of serotype O157:H7 possessed intimin type gamma1, and two strains of serotype O156:H- had the new intimin zeta. STEC O157:H7 strains were phage types 54 (four strains), 34 (two strains) and 14 (one strain). CONCLUSIONS: This study confirms that healthy sheep are a major reservoir of STEC pathogenic for humans. However, because the eae gene is present only in a very small proportion of ovine non-O157 STEC, most ovine strains may be less pathogenic. PMID- 12742716 TI - Uropathogenic virulence factors in isolates of Escherichia coli from clinical cases of canine pyometra and feces of healthy bitches. AB - Escherichia coli is commonly isolated in canine pyometra, but little is known of the virulence factors that may be involved in the precipitation of this disease. The aim of this study was to compare the prevalence of uropathogenic virulence factor (UVF) genes in E. coli isolates from canine pyometra and from feces of healthy bitches to evaluate their role in the pathogenesis of pyometra. E. coli from 23 cases of canine pyometra and from the feces of 24 healthy bitches were analyzed, by polymerase chain reaction, for UVF genes associated with canine and human urinary tract infections (UTIs). The prevalences of UVFs in E. coli from canine pyometra were similar to that in canine and human uropathogenic E. coli. The prevalence of pap was greater (P=0.036) for E. coli from pyometra (52%) than for fecal isolates (21%), and the papGIII allele was present in all pap containing isolates. The prevalences of genes for alpha-haemolysin and cytotoxic necrotising factor 1 were not significantly higher (P=0.075) in E. coli from pyometra than from feces. The proportion of pyometra strains with >or=3 UVFs was higher (P=0.039) than that of fecal strains, suggesting that possession of >or=3 UVF genes enhances the pathogenicity of the strain. Our findings demonstrate that E. coli associated with canine pyometra are similar to uropathogenic strains, and that operons that encode P fimbriae, alpha-haemolysin and cytotoxic necrotising factor 1 probably enhance the virulence and pathogenicity of the strain in the canine genital tract. PMID- 12742717 TI - Effect of freezing goat milk samples on recovery of intramammary bacterial pathogens. AB - With the aim of evaluating the effect of freezing goat milk samples on recovery of intramammary pathogens, 1200 milk samples from udder halves with subclinical intramammary infection were studied. Samples (20 ml) were frozen at -20 and at 80 degrees C. Thawing was carried out at room temperature at 7, 14, 21, 28, 58, 118, 178, 236 and 730 days after collection and bacteriological analyses were carried out to determine the number of colony forming units/ml (CFU/ml). Mixed model statistical analysis showed that bacterial group, temperature of storage, interaction of bacterial group and temperature of storage and the interaction of bacterial group, time and temperature of storage were statistically significant effects. For coagulase negative staphylococci (CNS), least squares means of log CFU/ml recovered at -20 and -80 degrees C were not different. Nevertheless, for Gram negative bacilli (GNB) a significant decrease was detected in samples frozen at -20 vs. -80 degrees C. At both temperatures and at different times of storage, significant increases were detected between log CFU/ml of CNS and values on day zero. At -20 degrees C, a significant decrease in GNB recovery was detected between freezing days zero and 730. This difference was not detected when goat milk samples infected by GNB were frozen at -80 degrees C. The results show that frozen milk samples can be useful in goat subclinical mastitis control programs. PMID- 12742718 TI - Iron acquisition by Actinobacillus suis: identification and characterization of transferrin receptor proteins and encoding genes. AB - Actinobacillus suis is an important pathogen of swine, especially in high-health status herds. A published report mentioning the binding of porcine transferrin (Tf) by at least one strain of A. suis suggested that A. suis, like other members of the Pasteurellaceae, can acquire Tf-bound iron by means of a siderophore independent, receptor-mediated mechanism. The objective of the present study was to characterize the components involved in this process, if present. Growth assays, with seven strains, confirmed that A. suis can use porcine (but not human or bovine) Tf as an iron source for growth. In solid phase binding assays, total membranes derived from all strains exhibited strong binding of porcine Tf, but only if the membranes were from organisms grown under iron-restricted conditions. An affinity-isolation procedure allowed the isolation of putative Tf-binding polypeptides ( approximately 100 and approximately 63 kDa) from comparable membranes from all strains. PCR approaches allowed the amplification, cloning and sequencing of A. suis tonB, exbB, exbD, tbpB and tbpA homologues. Reverse transcription (RT)-PCR, using RNA from organisms grown under iron-replete and iron-restricted conditions, revealed that tonB, exbB, exbD, tbpB and tbpA are transcribed as a single unit with expression being up-regulated in response to iron restriction. The calculated molecular masses of the predicted, mature TbpA (104.3 kDa) and TbpB (63.4 kDa) proteins suggest strongly that the affinity isolated, approximately 100 and approximately 63 kDa Tf-binding polypeptides represent TbpA and TbpB, respectively. It is concluded that the acquisition of Tf bound iron by A. suis involves mechanisms analogous to those found in other members of the Pasteurellaceae. PMID- 12742719 TI - Isolation of methylene dianiline and aqueous-soluble biodegradation products from polycarbonate-polyurethanes. AB - Polycarbonate-polyurethanes (PCNUs) have provided the medical device industry with practical alternatives to oxidation-sensitive polyether-urethanes (PEUs). To date, many studies have focused on PCNUs synthesized with 4,4'-methylene diphenyl diisocyanate (MDI). The relative hydrolytic stability of this class of polyurethanes is actually quite surprising given the inherent hydrolytic potential of the aliphatic carbonate group. Yet, there has been little information reporting on the rationale for the material's demonstrated hydrolytic stability. Recent work has shown that PCNU materials have a strong sensitivity towards hydrolysis when changes are made to their hard segment content and/or chemistry. However, knowledge is specifically lacking in regards of the identification of cleavage sites and the specific nature of the biodegradation products. Using high-performance liquid chromatography, radiolabel tracers and mass spectrometry, the current study provides insight into the distribution of biodegradation products from the enzyme-catalyzed hydrolysis of five different PCNUs. The hydrolytic sensitivity of the materials is shown to be related to the distribution of products, which itself is a direct consequence of unique micro structures formed within the different materials. While an MDI-based polymer was shown to be the most hydrolytically stable material, it was the only PCNU that produced its diamine analog, in this case 4,4'-methylene dianiline (MDA), as a degradation product. Given the concern over aromatic diamine toxicity, this finding is important and highlights the fact that relative biostability is a distinct issue from that of degradation product toxicity, and that both must be considered separately when assessing the impact of biodegradation on biomaterial in vivo compatibility. PMID- 12742720 TI - Activation of platelets adhered on amorphous hydrogenated carbon (a-C:H) films synthesized by plasma immersion ion implantation-deposition (PIII-D). AB - Amorphous carbon films have attracted much attention recently due to their good biocompatibility. Diamond-like carbon (DLC), one form of amorphous carbon that is widely used in many kinds of industries, has been proposed for use in blood contacting medical devices. However, the blood coagulation mechanism on DLC in a biological environment is not well understood. Platelet adhesion and activation are crucial events in the interactions between blood and the materials as they influence the subsequent formation of thrombus. In this work, the behavior of platelets adhered onto hydrogenated amorphous carbon films (a-C:H) is investigated. Hydrogenated amorphous carbon films with different hydrogen contents, structures, and chemical bonds were fabricated at room temperature using plasma immersion ion implantation-deposition (PIII-D). The wettability of the films was investigated by contact angle measurements using several common liquids. Platelet adhesion experiments were conducted to examine the interaction of blood with the films in vitro and the activation of adherent platelets. The results show that the behavior of the platelets adhered on the a-C:H films is influenced by their structure and chemical bond, and it appears that protein interaction plays a key role in the activation of the adherent platelets. PMID- 12742721 TI - Enzyme-catalyzed gel formation of gelatin and chitosan: potential for in situ applications. AB - We compared the ability of two enzymes to catalyze the formation of gels from solutions of gelatin and chitosan. A microbial transglutaminase, currently under investigation for food applications, was observed to catalyze the formation of strong and permanent gels from gelatin solutions. Chitosan was not required for transglutaminase-catalyzed gel formation, although gel formation was faster, and the resulting gels were stronger if reactions were performed in the presence of this polysaccharide. Consistent with transglutaminase's ability to covalently crosslink proteins, we observed that the transglutaminase-catalyzed gelatin chitosan gels lost the ability to undergo thermally reversible transitions (i.e. sol-gel transitions) characteristic of gelatin. Mushroom tyrosinase was also observed to catalyze gel formation for gelatin-chitosan blends. In contrast to transglutaminase, tyrosinase-catalyzed reactions did not lead to gel formation unless chitosan was present (i.e. chitosan is required for tyrosinase-catalyzed gel formation). Tyrosinase-catalyzed gelatin-chitosan gels were observed to be considerably weaker than transglutaminase-catalyzed gels. Tyrosinase-catalyzed gels were strengthened by cooling below gelatin's gel-point, which suggests that gelatin's ability to undergo a collagen-like coil-to-helix transition is unaffected by tyrosinase-catalyzed reactions. Further, tyrosinase-catalyzed gelatin-chitosan gels were transient as their strength (i.e. elastic modulus) peaked at about 5h after which the gels broke spontaneously over the course of 2 days. The strength of both transglutaminase-catalyzed and tyrosinase-catalyzed gels could be adjusted by altering the gelatin and chitosan compositions. Potential applications of these gels for in situ applications are discussed. PMID- 12742723 TI - Preparation of hydroxyapatite-gelatin nanocomposite. AB - A nanocomposite of gelatin[GEL]-hydroxyapatite[HAp] was prepared using the biomimetic process. The hydroxyapatite nanocrystals were precipitated in aqueous solution of gelatin at pH 8 and 38 degrees C. The chemical bonding between calcium ions of HAp and carboxyl ions of GEL molecules induced a red-shift of the 1339 cm(-1) band of GEL in FT-IR analysis. TEM images and electron diffraction patterns for the nanocomposite strongly indicate the self-organization of HAp nanocrystals along the GEL fibrils. Electron diffraction for the nanocomposites showed a strong preferred orientation of the (002) plane in HAp nanocrystals. The development of HAp nanocrystals in an aqueous GEL solution was highly influenced by the concentration ratio of GEL to HAp. A higher concentration of GEL induced the formation of tiny crystallites (4 nm x 9 nm size), while a lower concentration of GEL contributed to the development of bigger crystallites (30 nm x 70 nm size). From DT/TGA data, the HAp-GEL nanocomposite showed typically three exothermic temperatures. The increase in decomposition temperatures indicates the formation of a primary chemical bond between HAp and GEL. The higher concentration of GEL supplies abundant reaction sites containing groups such as carboxyl, which can bind with calcium ions. The abundant supply of reaction sites leads to a very large number of HAp nuclei. However, the formation of a large number of nuclei depletes the concentration of calcium ions that available for growth to the extent that the nuclei cannot grow very large. This in turn will lead to the creation of a large number of tiny nanocrystals at this higher GEL concentration. PMID- 12742722 TI - Adhesion, apoptosis and cytokine release of human mononuclear cells cultured on degradable poly(urethane urea), polystyrene and titanium in vitro. AB - Early interactions between materials and mononuclear cells may influence the viability and secretory response of the cells. Such effects may in turn influence the subsequent inflammatory and repair phases around the materials. In the present study, it was examined if mononuclear cells cultured in vitro either unstimulated or stimulated with lipopolysaccharide (LPS) (10ng/ml) revealed differences regarding cell viability and apoptosis. A major interest was to study the influence of different material properties on the parameters of the inflammatory response upon cell adhesion to materials with widely different surface chemical properties but similar surface topography: degradable poly(urethane urea) (PUUR), cell culture treated polystyrene (PS) surfaces, and commercially pure (c.p.) titanium (Ti). Finally, the secretion of the proinflammatory tumor necrosis factor-a (TNF-alpha) and the downregulating interleukin-10 (IL-10) cytokines was examined in the supernatants from 24h mononuclear cell cultures. No differences in cell viability as measured by lactate dehydrogenas (LDH) were observed between the three materials. The number of material-surface adherent cells was higher on PUUR than the more hydrophilic PS and Ti as judged by quantification of material surface-associated DNA, light microscopic morphological examination of DAPI-stained cells and SEM. LPS increased the number of adherent cells, irrespective of the type of material. The lowest number of apoptotic (annexin-V) and necrotic (propidium iodide) mononuclear cells was detected on PUUR. LPS decreased the number of both apoptotic and necrotic cells, irrespective of material. Low TNF-alpha levels were detected in unstimulated conditions, irrespective of material types. A significantly lower amount of TNF-alpha was found with unstimulated cells on PUUR than on Ti. A significantly higher IL-10 level was detected in unstimulated Ti cultures compared with PUUR and PS. Secretion of IL-10 was predominantly stimulated by LPS on PUUR and Ti. The data indicate that material-related differences are expressed in differences in cell adherence, apoptosis and cytokine secretion. Further, degradable PUUR has equal or less cell-activating properties than Ti and PS under in vitro conditions. PMID- 12742724 TI - A modified microstamping technique enhances polylysine transfer and neuronal cell patterning. AB - Macromolecular microstamping with polydimethylsiloxane (PDMS) stamps has been demonstrated to transfer proteins onto glassy substrates for antigen or antibody detection and for cell patterning. For many applications, including neuronal cell patterning, it is important to assure reliable transfer of sufficient quantity of protein. Research has shown that protein transfer is enhanced with the selection of the proper protein-stamp-substrate combination. In addition, detergent studies have shown that detergent-protein complexes detach from surfaces to a greater extent than proteins alone. Therefore, we hypothesized that stamp surface modification (termed here a release layer) can enhance polylysine transfer and benefit cell growth on microstamped substrates. We found unmodified stamps to transfer insufficient polylysine to support good cell survival of hippocampal neurons in a widely used serum-free, reduced-glia cell culture system. However, with modified stamps neuronal growth was reliably good. This enhanced cell growth can be attributed to the increased polylysine transfer due to the release layer rather than increased loading onto the stamp. This enhancement was found to be even greater for two-month old stamps that were stored in water. Furthermore, the physicochemical properties of the release layer can modulate the loading process. Thus, our data supports the conclusions that the release layer: (1) modulates polylysine loading, (2) enhances polylysine transfer, (3) enhances cellular growth on microstamped substrates, and (4) extends the durability (defined as the number of times a stamp can be reused) of PDMS microstamps. PMID- 12742725 TI - Study on physical properties and nerve cell affinity of composite films from chitosan and gelatin solutions. AB - A series of chitosan-gelatin composite films was prepared by varying the ratio of constituents. FT-IR and X-ray analysis showed good compatibility between these two biopolymers. Differential scanning calorimetry (DSC) analysis indicated that the water take-up of chitosan film increased when blended with gelatin. Composite film exhibited a lower Young's modulus and a higher percentage of elongation-at break compared with chitosan film, especially in wet state. All composite films were hydrophilic materials with water contact angles ranging from 55 degrees to 65 degrees. The results obtained from ELISA indicated the adsorption amount of fibronectin on composite films was much higher than on chitosan film. PC12 cells culture was used to evaluate the nerve cell affinity of materials. The cells cultured on the composite film with 60wt% gelatin differentiated more rapidly and extended longer neurites than on chitosan film. The results suggest that the soft and elastic complex of chitosan and gelatin, which has better nerve cell affinity compared to chitosan, is a promising candidate biomaterial for nerve regeneration. PMID- 12742727 TI - Preparation of alginic acid layers on stainless-steel substrates for biomedical applications. AB - This study is concerned with the blood compatibility of alginic acid layers immobilized on gamma-aminopropyltriethoxysilane (gamma-APS)-grafted stainless steel (SUS316L). The surfaces were characterized with contact angle measurement and X-ray photoelectron spectroscopy (XPS). The blood compatibility was evaluated in terms of platelet adhesion and blood clotting time. An in vitro platelet adhesion assay indicated that only a small number of platelets adhered to substrate surfaces modified with gamma-APS and subsequently with alginic acid. Moreover, alginic-acid-immobilized SUS316L substrates had little effect on the blood clotting time. This indicated that alginic-acid-immobilized SUS316L substrates do not adsorb some blood-clotting proteins or factors, or stimulate them. PMID- 12742726 TI - Effect of bifunctional comonomers on mechanical strength and water sorption of amorphous calcium phosphate- and silanized glass-filled Bis-GMA-based composites. AB - This study seeks to elucidate structure-property relationships in a series of unfilled dental copolymers and their composites. The copolymers/composites were derived from photo-activated binary monomer systems based on 2,2-bis[p-2'-hydroxy 3'-methacryloxypropoxy)phenyl] propane (Bis-GMA) and equimolar amounts of a bifunctional, surface-active comonomer, i.e., 2-hydroxyethyl methacrylate (HEMA), glycerol dimethacrylate (GDMA) or ethylene glycol methacrylate phosphate (PHEMA). Triethyleneglycol dimethacrylate, a widely used comonomer for Bis-GMA, was used as a control. Two types of fillers were investigated: (1) a hydrophilic, silica modified amorphous calcium phosphate (Si-ACP) and (2) a more hydrophobic, silanized nanosized silica (n-SiO(2)). Both the unfilled copolymers and their composites were evaluated for biaxial flexure strength (BFS), both dry and wet after 30 days immersion in buffered saline, and for water sorption (WS) and their WS kinetic profiles. The Bis-GMA copolymers and composites derived from HEMA and GDMA had BFS and WS values, as well as WS kinetic profiles, similar to the controls. Copolymers and composites based on Bis-GMA/PHEMA had lower BFS and higher WS values. Si-ACP composites had significantly lower BFS values (that were further diminished on soaking) than their copolymers. WS increased as the level of this filler was increased except for Bis-GMA/PHEMA composites. With n-SiO(2) as the filler, a more moderate reduction in BFS occurred compared to the unfilled copolymers. By contrast to Si-ACP composites, the WS of all the n-SiO(2) composites decreased with increasing filler level. From this study it is evident that both the chemical structure of the polymer matrix and the type of filler system can have significant effects on the strength and water-related properties of dental composites. PMID- 12742728 TI - UHMWPE oxidation increases granulocytes activation: a role in tissue response after prosthesis implant. AB - Ultra-high molecular weight polyethylene (UHMWPE), a biopolymer widely used in orthopaedic implants, is oxidized during gamma-ray sterilization; such surface oxidation is considered as major responsible for inflammation and prosthesis failure. As granulocytes are involved in first contact inflammation, we have measured their oxidative burst by flow cytometry using dihydrorhodamine 123 (DRH) to evaluate their activation following contact with normal and oxidized UHMWPE. Peripheral blood cells (obtained by lysed blood) were loaded with DRH, seeded onto polystyrene, normal and heat-oxidized UHMWPE disks for 30min and then collected for analysis. Granulocytes were individuated using FSC and SSC signals and their cell associated green fluorescence was analyzed. Both normal and oxidized UHMWPE stimulated granulocytes activation as showed by the mean fluorescence emitted (109.3+/-3.8 and 150.1+/-9.2, respectively) compared to control samples (81.6+/-0.3). Moreover oxidized UHMWPE activated a significantly higher percentage of granulocytes (73.35+/-5.2%) compared to not-oxidized UHMWPE (21.5+/-3.8%). UHMWPE surface oxidation responsible for increased granulocyte activation seems to play a role in tissue response to implants. PMID- 12742729 TI - Reactive fibre reinforced glass ionomer cements. AB - The mechanical properties of glass ionomer cements used in restorative dentistry reinforced by chopped glass fibres were investigated. Reactive glass fibres with a composition in the system SiO(2)-Al(2)O(3)-CaF(2)-Na(3)AlF(6) and a thickness of 26 microm were drawn by a bushing process. The manufacturing parameters were optimized with respect to maximum strength of the glass fibre reinforced ionomer cements. Powder to liquid ratio, pre-treatment of the glass, grain size distribution and fibre volume fraction were varied. Glass fibre and cement were characterized by X-ray diffraction, transmission electron microscopy and energy dispersive spectroscopy techniques, respectively. The highest flexural strength of the reinforced cement (15.6 MPa) was found by compounding 20 vol% reactive fibres and extending the initial dry gelation period up to 30 min. Microscopic examination of the fractured cements indicated a distinct reactive layer at the fibre surface. A pronounced fibre pull out mode gives rise to an additional work of-fracture contributed by pulling the fibres out of the fracture surface. PMID- 12742730 TI - Assessment of genetic damage by methyl methacrylate employing in vitro mammalian test system. AB - Methyl methacrylate (MMA) is a volatile liquid widely used in the manufacture of acrylic polymers. In modern dentistry, MMA is the mainstream material in denture bases. MMA has been implicated as primary irritant and sensitizer, which can cause allergic eczematous reaction on the oral mucosa as well as skin. To date, there is growing concern that MMA may produce genetic damage by inducing mutation. In this study, colony forming efficiency, DNA synthesis, and cytogenetic assays were performed to investigate the adverse effects of MMA in cultured CHO cells. MMA was found to decrease colony formation in a dose- and time-dependent manner (P<0.05). MMA also inhibited DNA synthesis in a dose dependent manner (P<0.05). The chromosome aberrations induced by MMA were the chromatid-type aberrations in the treated cultures. Moreover, the gaps and breaks were the most common type of aberrations observed. The sister-chromatid exchange frequencies were found to increase in the concentration of MMA. In this study, MMA was found to be not only a cytotoxic agent but also a genotoxic agent. The effects observed following treatment with low dose for longer duration is of relevance to the condition of the oral mucosa of the denture wears. Denture base resin could constantly release MMA extended periods, possibly causing moderate toxic reactions and possibly contributing to adverse effects on the mucosa. PMID- 12742731 TI - Nano-structured polymers enhance bladder smooth muscle cell function. AB - It is the hypothesis of the present study that a biocompatible material which mimics the nanometer topography of native bladder tissue will enhance cellular responses and lead to better tissue integration in vivo. Previous in vitro studies have verified the ability to successfully reduce the surface feature dimensions of poly(lactic-co-glycolic acid) (PLGA) and poly(ether urethane) (PU) films into the nanometer regime via chemical etching procedures. Results from these studies also provided the first evidence that bladder smooth muscle cell adhesion was enhanced on chemically treated nano-structured polymeric surfaces compared to their conventional counterparts. Although cell adhesion is necessary for a biomaterial's success, subsequent cell functions (such as long-term cell growth and proliferation) are also critical for tissue ingrowth and long-term implant survival. The present in vitro study, therefore, investigated the function of bladder smooth muscle cells on these novel, nano-structured polymers over the expanded periods of 1, 3 and 5 days. Results indicated that cell number was influenced by both surface roughness and surface chemistry changes; the important contributor, however, was increased nanometer surface roughness. This claim is supported by the fact that cell number was enhanced on nano-structured compared to conventional PLGA and PU once chemistry changes were eliminated using casting techniques. PMID- 12742732 TI - TAK-778 enhances osteoblast differentiation of human bone marrow cells cultured on titanium. AB - TAK-778 induces bone growth in in vitro and in vivo models. The aim of this study was to evaluate the osteogenic potential of TAK-778 on human bone marrow cells cultured on commercially pure titanium (cpTi). Cells were cultured either in absence or in presence of TAK-778 (10(-5)M) on cpTi in supplemented alpha-MEM. For attachment evaluation, cells were cultured for 4 and 24h. After 7, 14, and 21 days, cell proliferation, cell viability, total protein content, alkaline phosphatase (ALP) activity, and bone-like formation were evaluated. TAK-778 did not affect cell attachment and viability. Cell number was reduced by TAK-778. ALP activity, total protein content, and bone-like formation were increased by TAK 778. These results suggest that initial cell events such as cell attachment are not affected by TAK-778 while events that indicate osteoblast differentiation including reduced cell proliferation, and increased both ALP activity and bone like formation are enhanced by TAK-778 in presence of cpTi. It means that TAK-778 could be a useful drug to improve the osseointegration of implants by both enhancing and accelerating bone formation on Ti surface. PMID- 12742733 TI - Shrinkage stresses in bone cement. AB - Shrinkage of bone cement is reported primarily as a consequence of polymerisation, however thermal shrinkage also occurs as a result of its exothermic reaction. It is proposed that the latter effect is important, since it occurs late in the curing cycle at a time when the cement has attained its mechanical properties as a solid, and that residual stresses result. Observations from experiments and literature reports suggest that residual stresses may be sufficient to initiate cracks at the interface between hip replacement stems and cement.A theoretical model has been developed to calculate interference stresses, using thick-walled cylinder theory, on the basis of thermal and total shrinkages. Thermal shrinkage values were calculated using the coefficient of linear thermal expansion of bone cement, while total shrinkages were measured. Moduli of elasticity values were measured for acrylic bone cements ranging from 2.1 to 2.7GPa, as were Poisson's ratio values ranging from 0.38 to 0.46. Theoretical calculation of stresses in a cement mantle, based on assumptions of thermal shrinkage alone, predicted circumferential stresses of 8.4-25.2MPa for cement curing temperatures in the range 60-140 degrees C. It is concluded that cracks observed around hip prosthesis stems in laboratory specimens of bone cement are due to shrinkage and that residual stresses are sufficient to cause crack initiation prior to functional loading. PMID- 12742734 TI - Fabrication of a new composite orthodontic archwire and validation by a bridging micromechanics model. AB - A new technique based on tube shrinkage is proposed for the fabrication of composite archwires. Compared with a traditional pultrusion method, this new technique can avoid any fiber damage during the fabrication and can provide the archwire with a required curvature in its final clinical usage. The present paper focuses on the technique development and mechanical design and validation in terms of constituent materials by using a micromechanics bridging model. Prototype archwire has been fabricated using fiberglass and an epoxy matrix, with a wire diameter of 0.5mm and a 45% fiber volume fraction. Tensile and three-point bending tests have shown that the mechanical performance of the prototype composite archwire is comparable to that of a clinical Ni-Ti archwire. Another purpose of the present paper is to provide an efficient procedure for a critical design of composite archwires. For this to be possible, the ultimate load especially flexural load carrying ability of the composite archwire must be assessed from the knowledge of its constituent properties. However, difficulty exists in doing this, which comes from the fact that the failure of the utmost filament of the composite archwire subjected to initially the maximum bending stress does not imply its ultimate failure. Additional higher loads can still be applied and a progressive failure process is generated. In this paper, the circular archwire was discretized into a number of parallel laminae along its axis direction, and the bridging micromechanics model combined with the classical lamination theory has been applied to understand the progressive failure process with reasonable accuracy. Only the constituent fiber and matrix properties are required for this understanding. Nevertheless, the ultimate bending strength cannot be obtained only based on a stress failure criterion. This is because neither the first-ply nor the last-ply failure corresponds to the ultimate failure. An additional critical deflection (curvature) condition must be employed also. By using both the stress failure and the critical deflection conditions, the predicted load-deflection up to the ultimate failure agrees well with the measured data. Thereafter, different mechanical performances of composite archwires can be tailored before fabrication by choosing suitable constituent materials, their contents, and the archwire diameters. Several design examples have been shown in the paper. PMID- 12742735 TI - Preparation of ceramic microspheres for in situ radiotherapy of deep-seated cancer. AB - Radiotherapy is one of the most effective treatments for cancers. However, external irradiation provides only small doses to deep-seated cancers, and often causes damage to healthy tissues. It has been reported that 20-30 microm diameter 17Y(2)O(3)-19Al(2)O(3)-64SiO(2) (mol%) glass microspheres are useful for the in situ irradiation of cancers. Yttrium-89 (89Y) in this glass can be neutron bombarded to form the beta-emitter 90Y (half-life=64.1h). When injected in the vicinity of the cancer, such activated glass microspheres can provide a large localized dose of beta-radiation. The Y(2)O(3) content of the glass in the microspheres is limited to only 17 mol%. Chemically durable microspheres with a higher Y(2)O(3) content need to be developed. Phosphorus-31 (31P) with 100% natural abundance can also be activated by neutron bombardment to form the beta emitter 32P (half-life=14.3d). Chemically durable microspheres containing a high phosphorus content are expected to be more effective for cancer treatment. We prepared pure Y(2)O(3) and YPO(4) microspheres using a high-frequency induction thermal plasma melting technique, and investigated the resulting structure and chemical durability. We successfully prepared smooth, highly spherical polycrystalline Y(2)O(3) and YPO(4) microspheres with diameters in the range 20 30 microm. Both the Y(2)O(3) and YPO(4) microspheres showed high chemical durability in saline solutions buffered at pH=6 and 7. These microspheres are expected to be more effective than the conventional glass microspheres for the in situ radiotherapy of cancer. PMID- 12742736 TI - Microstructural modifications induced by the entrapped glucose oxidase in cross linked polyacrylamide microgels used as glucose sensors. AB - Glucose oxidase (GO(x)) has been immobilized in cross-linked polyacrylamide microgels using the concentrated emulsion polymerization method. The immobilization of the enzyme is effective when the number of cross-links is equal or greater than 0.021mol/g. The swelling of the microgels is not affected by the entrapped enzyme but decreases with increasing the number of cross-links. The glass transition temperature of microgels with GO(x) presents considerable differences with that of the empty microgels, which are explained on base of the competition between the plasticizer effect of the encapsulated enzyme and the chain mobility restrictions imposed by the microgel cross-linking content. Immobilization of GO(x) slightly modifies the X-ray diffraction patterns of the microgels; however, the diffraction profiles of cross-linked microgels suggest the formation of bis(acrylamide) aggregates in the outer layers of the microgel particles. PMID- 12742737 TI - Hyperthermia following traumatic brain injury: a critical evaluation. AB - Hyperthermia, frequently seen in patients following traumatic brain injury (TBI), may be due to posttraumatic cerebral inflammation, direct hypothalamic damage, or secondary infection resulting in fever. Regardless of the underlying cause, hyperthermia increases metabolic expenditure, glutamate release, and neutrophil activity to levels higher than those occurring in the normothermic brain-injured patient. This synergism may further compromise the injured brain, enhancing the vulnerability to secondary pathogenic events, thereby exacerbating neuronal damage. Although rigorous control of normal body temperature is the current standard of care for the brain-injured patient, patient management strategies currently available are often suboptimal and may be contraindicated. This article represents a compendium of published work regarding the state of knowledge of the relationship between hyperthermia and TBI, as well as a critical examination of current management strategies. PMID- 12742738 TI - Intact spatial memory in mice with seizure-induced partial loss of hippocampal pyramidal neurons. AB - We generated defined neuronal loss in hippocampus of genetically identical mice by pilocarpine injections and studied the impact of these seizures on the performance of mice in spatial learning and memory. The numbers of TUNEL-positive degenerating cells paralleled the severity of the seizures. When compared to the numbers found for not-seizured control mice, mild, moderate, and severe seizures produced significant increases in TUNEL-positive neurons in CA1 and CA3 regions by 19, 25, and 63%, respectively. Water maze learning was abolished after the severe seizures. However, spatial learning was normal after mild or moderate seizures. Therefore, there was no linear correlation between the impairment of learning and memory performance with the number of degenerating neurons in hippocampus. Our data suggest that normal spatial learning and memory can be achieved without the full number of hippocampal pyramidal neurons in partially lesioned hippocampus. PMID- 12742739 TI - Fas and Fas ligand are associated with neuritic degeneration in the AD brain and participate in beta-amyloid-induced neuronal death. AB - It has recently been suggested that neuronal cell death in response to many brain insults may be mediated by the upregulation of tumor necrosis factor receptor (TNFR) family members and their ligands. In the present study, we investigated whether the expression of the TNFR family death domain receptor, Fas, and its ligand, FasL, is altered in association with neuropathology and activated caspase markers in Alzheimer disease (AD) brain, and Abeta-induced neuronal cell death in vitro. To evaluate this hypothesis, we examined Fas and FasL expression in AD and control brain, and Abeta-treated primary neurons, using immunocytochemistry and Western blots. Neurons in both AD brain and Abeta-treated cultures exhibited FasL upregulation and changes in immunoreactivity for Fas receptor. Further, FasL expression was remarkably elevated in senile plaques and neurofilament-positive dystrophic neurites, and in association with caspase activation and neuritic apoptosis in AD brain. Based on these and previous data regarding protection of primary neuronal cultures from Abeta(1-42)-induced apoptosis by blockade of Fas associated death domain signaling, we also tested the hypothesis that dynamic regulation of Fas and FasL may contribute to Abeta-mediated neuronal cell death. Accordingly, neuronal cultures derived from mice carrying inactivating mutations in Fas (Faslpr) or FasL (Fasgld) exhibited protection from Abeta(1-42)-induced cell death. These findings suggest that Fas-FasL interactions may contribute to mechanisms of neuronal loss and neuritic degeneration in AD. PMID- 12742740 TI - Normal cognitive behavior in two distinct congenic lines of transgenic mice hyperexpressing mutant APP SWE. AB - Amyloid deposition appears to be an early and crucial event in Alzheimer's disease (AD). To generate animal models of AD, mice expressing full-length amyloid precursor protein (APP), with mutations linked to FAD, have been created. These animals exhibit abnormalities characteristic of AD, including deposits of beta-amyloid (Abeta), neuritic plaques, and glial responses. In studies of cognition in these animals, there have been several reports of memory disturbances well before the appearance of amyloid deposits. We have developed two distinct lines of transgenic mice (C3-3 and E1-2) that express the "Swedish" variant of APP (APP(SWE)) at levels that are approximately three-fold higher than endogenous mouse APP. Both lines have been backcrossed to C57BL/6J mice for 10 generations. Here, we use longitudinal and cross-sectional studies to evaluate the cognitive performance of our animals, where the concentration of Abeta1-42 in brain increases with aging from low levels (2-10 pmol/g) at 6-14 months of age to relatively high levels (60-100 pmol/g) at 24-26 months, when deposits of Abeta were beginning to form. When 12-month-old mice were tested in tasks that assess reference and working memory, transgenic mice from both lines could not be distinguished from nontransgenic littermates. Further study of 24- to 26-month old transgenic mice (C3-3 line) found no evidence of memory impairment despite the presence of high levels of human Abeta (60-100 pmol/g). Thus, the expression of APP(SWE) at approximately three-fold over endogenous levels, which is sufficient to induce amyloid deposition at advanced ages, does not significantly erode cognitive performance in aged mice. PMID- 12742741 TI - Enhanced long-term potentiation in the hippocampus of rats expressing mutant presenillin-1 is age related. AB - Electrophysiological recordings were made from Fischer rats engineered to express the human presenilin 1 gene carrying the M146V mutation. Extracellular recordings of field excitatory post-synaptic potential (EPSPs) were made to investigate EPSP properties, paired pulse responses, posttetanic potentiation, and long-term potentiation in the stratum radiatum and dentate gyrus of hippocampal slices maintained in vitro. Transgenic rats aged approximately 6 months showed no differences from their wild-type littermates in any of these properties. However, at 18 months, long-term potentiation in the CA1 was facilitated in the transgenic rats with a different pattern of synaptic enhancement. No changes were observed in paired pulse facilitation (PPF) or post-tetanic potentiation (PPT) and no changes were seen in the dentate gyrus. Field potential amplitudes were significantly greater and PPF was enhanced in the CA1 of all older rats. Intracellular recordings from CA1 pyramidal cells of the older group of rats revealed no differences in the passive or active membrane properties of cells in the two groups, but intracellularly recorded EPSPs were significantly longer. PMID- 12742742 TI - Molecular phylogeny and phylogeography of free-living Bryozoa (Cupuladriidae) from both sides of the Isthmus of Panama. AB - Genetic data were used to identify Recent species of free-living bryozoans (Cupuladriidae) from both sides of the Isthmus of Panama, and to examine their phylogenetic relationships, species richness, and population structures. An approximately 480bp fragment of the 16S mitochondrial rRNA gene was sequenced from 182 individuals from Panama, the Gulf of Mexico, and El Salvador. Ten haplotype groups (Cupuladria 4, 5, and 6; Discoporella 1, 2, 3A, 3B, 3C, 7, and 8) were identified. Genetic distances between haplotype groups (3.2-26.5%; K2P+Gamma) were 1-2 orders of magnitude greater than within groups (0.1-1.4%). Seven of the haplotype groups represent morphologically distinct species; Discoporellas 3A-C appear to be cryptic species. Phylogenetic analyses identified two pairs of transisthmian sister clades. An average divergence rate derived from other taxa suggests that Cupuladrias 4 and 5 diverged approximately 7Ma, a Discoporella 7 clade diverged from a 3A-C clade approximately 11Ma, and the 3A-C clade radiated approximately 6-4Ma; these events all predated final closure of the isthmus? 3Ma. The Caribbean side of the isthmus, with 5 species, is only marginally richer in cupuladriids than the Pacific side, with 4, but has greater phylogenetic depth. The Caribbean retains lineages stemming from a New World Miocene radiation that are not represented in the eastern Pacific; extant eastern Pacific cupuladriids share most recent common ancestry with only two of the Caribbean lineages. Species in the eastern Pacific tend to show shallow population structures, with high levels of gene flow between geographically separate populations, whereas Caribbean species tend to show deeper populations structures, with indications of restricted gene flow between Bocas del Toro/Gulf of Mosquitos and Costa Arriba/San Blas. The population structures derive from Pleistocene histories and may be of limited value in interpreting the macroevolutionary pattern, as our results provide no evidence of speciation on either side of the isthmus following closure in the late Pliocene. PMID- 12742743 TI - Phylogenetic significance of morphological characters in the taxonomy of Pestalotiopsis species. AB - There has been considerable disagreement regarding the relationships among Pestalotiopsis species and their delimitations. A molecular phylogenetic analysis was conducted on 32 species of Pestalotiopsis in order to evaluate the utility of morphological characters currently used in their taxonomy. Phylogenetic relationships were inferred from nucleotide sequences in the ITS regions and 5.8S gene of the rDNA under four optimality criteria: maximum parsimony, weighted parsimony, maximum likelihood, and neighbor joining. Phylogenies estimated from all analyses yielded trees of essentially similar topology and revealed 3 major groups that correspond with morphology-based classification systems. Molecular data indicated that the genus contains two distinct lineages based on pigmentation of median cells and four distinct groupings based on morphology of apical appendages. The analyses did not support reliability of other phenotypic characters of this genus, such as spore dimensions. Characters with particular phylogenetic significance are discussed in relation to the taxonomy of Pestalotiopsis. PMID- 12742744 TI - A phylogeny of the Australian Sphenomorphus group (Scincidae: Squamata) and the phylogenetic placement of the crocodile skinks (Tribolonotus): Bayesian approaches to assessing congruence and obtaining confidence in maximum likelihood inferred relationships. AB - Australian scincid lizards are a diverse squamate assemblage ( approximately 385 species), divided among three major clades (Egernia, Eugongylus, and Sphenomorphus groups). The Sphenomorphus group is the largest, comprising 61% of the Australian scincid fauna. Phylogenetic relationships within the Australian Sphenomorphus group and the phylogenetic placement of Tribolonotus are inferred using mtDNA (12S and 16S rRNA genes, ND4 protein-coding gene, and associated tRNA genes; 2185bp total). These data were analyzed separately (structural RNA vs protein-coding partitions) and combined using maximum likelihood. Confidence in inferred clades was assessed using non-parametric bootstrapping and Bayesian analysis. Analysis of the combined data strongly supports Sphenomorphus group (as well as the Australian subgroup) monophyly. Notoscincus is strongly placed as the sister taxon of the remaining Australian Sphenomorphus group taxa, with this more exclusive clade being divided into two major groups (one restricted to mesic eastern Australia and the other continent wide). The speciose Australian "Eulamprus" and "Glaphyromorphus" are both polyphyletic. All remaining non Sphenomorphus group lygosomine skinks strongly form a clade, with Tribolonotus placed as the sister taxon of the Australian Egernia group. PMID- 12742745 TI - A phylogeny of Chinese species in the genus Phrynocephalus (Agamidae) inferred from mitochondrial DNA sequences. AB - We investigated the phylogenetic relationships among most Chinese species of lizards in the genus Phrynocephalus (118 individuals collected from 56 populations of 14 well-defined species and several unidentified specimens) using four mitochondrial gene fragments (12S rRNA, 16S rRNA, cytochrome b, and ND4 tRNA(LEU)). The partition-homogeneity tests indicated that the combined dataset was homogeneous, and maximum-parsimony (MP), neighbor-joining (NJ), maximum likelihood (ML) and Bayesian (BI) analyses were performed on this combined dataset (49 haplotypes including outgroups for 2058bp in total). The maximum parsimony analysis resulted in 24 equally parsimonious trees, and their strict consensus tree shows that there are two major clades representing the Chinese Phrynocephalus species: the viviparous group (Clade A) and the oviparous group (Clade B). The trees derived from Bayesian, ML, and NJ analyses were topologically identical to the MP analysis except for the position of P. mystaceus. All analyses left the nodes for the oviparous group, the most basal clade within the oviparous group, and P. mystaceus unresolved. The phylogenies further suggest that the monophyly of the viviparous species may have resulted from vicariance, while recent dispersal may have been important in generating the pattern of variation among the oviparous species. PMID- 12742746 TI - Phylogenetic relationships within the coral crab genus Carpilius (Brachyura, Xanthoidea, Carpiliidae) and of the Carpiliidae to other xanthoid crab families based on molecular sequence data. AB - The coral crab genus Carpilius currently includes three widely distributed species that inhabit tropical coral reefs and adjacent waters. The relationship of Carpilius to other xanthoid crabs is unknown. Previously, carcinologists considered Carpilius to be allied with crabs of the family Xanthidae (e.g., Euryozius, Liagore, and Liomera), however, recent workers have considered it to be a monotypic genus within its own family, Carpiliidae. Mitochondrial 12S- and 16S-rDNA gene fragments confirm the monophyly and distinct status of the family Carpiliidae. Within the genus Carpilius, the Caribbean species C. corallinus is basal to the two Pacific species C. maculatus and C. convexus. The Pacific species are sister taxa, despite the greater morphological resemblance of C. corallinus to the Pacific C. convexus. The relationship of the Carpiliidae (Carpilius) to other xanthoid crabs is investigated, and results of a preliminary analysis of higher xanthoid relationships did not resolve the relationships of Carpiliidae, "Xanthidae," Menippidae, Trapeziidae, and Ocypodidae to one another. A Menippidae and Carpilius relationship could not be rejected, although a Liomera, Liagore, and Carpilius relationship was. PMID- 12742747 TI - Diversification of sympatric Sapromyza (Diptera: Lauxaniidae) from Madeira: six morphological species but only four mtDNA lineages. AB - A series of recent studies on speciation of insects within the Canary Islands have indicated considerable within-island diversification, similar to that described in the Hawaiian islands. Little work has yet been carried out on the neighboring Madeiran archipelago, which is also volcanic. This study examines relationships among all known Lauxaniid flies of the genus Sapromyza from Madeira (including six newly described morphological species) based on mitochondrial gene trees constructed from cytochrome c oxidase (subunit I) and 16S rRNA partial sequences. Phylogenies based on maximum likelihood distances, a Bayesian method based on Markov chain Monte Carlo sampling from the posterior probability distribution, and maximum parsimony show that eight of the nine Madeiran species comprise a single monophyletic group. This clade is also split into two subclades representing black- and yellow/orange-bodied forms. The latter mtDNA clade corresponds to only two species (Sapromyza imitans and Sapromyza indigena) which are not reciprocally monophyletic. Monophyly is strongly supported within four of the six black-bodied species but not for the species pair (Sapromyza inconspicua, Sapromyza laurisilvae). We discuss the double occurrence (at least) of introgressive hybridization/incomplete lineage sorting within this group and suggest that recent speciation is the most likely explanation. The remaining species on the island, Sapromyza madeirensis, is very divergent from the aforementioned group, occupying a more basal position in the tree than the other Atlantic island and continental Sapromyza that were included in the analysis. At least two speciation events for Madeiran Sapromyza appear to correspond to quite ancient periods relative to the age of the island, while others are more recent. This suggests that a combination of island colonization and within-island sympatric and/or vicariance-mediated speciation may explain the observed diversity. PMID- 12742748 TI - Phylogenetic analysis of the subclass Pteriomorphia (Bivalvia) from mtDNA COI sequences. AB - Phylogenetic relationships within the subclass Pteriomorphia (Bivalvia) were examined using sequences of the mitochondrial cytochrome c oxidase subunit I gene. The resultant Minimum Evolution phylogenetic tree strongly supports the existing superfamily-level classification with all the members of each superfamily forming clades. At the same time, it is suggested that: (1) Ostreoidea shows a closer relationship to Pinnoidea and Pterioidea than to the other superfamilies; (2) Pectinoidea, Anomioidea, and Limoidea form a clade, (3) Arcoidea and Limopsoidea form a clade; (4) The subclass Anomalodesmata is closer to the subclass Heterodonta than to Mytiloidea; and (5) The subclass Pteriomorphia is monophyletic. Taking these results as well as published data for nuclear 18S ribosomal DNA (18S rDNA) and Myosin analyses into consideration, a new order-level classification system for Pteriomorphia is proposed. PMID- 12742749 TI - Molecular phylogeny of Crematogaster subgenus Decacrema ants (Hymenoptera: Formicidae) and the colonization of Macaranga (Euphorbiaceae) trees. AB - To elucidate the evolution of one of the most species-rich ant-plant symbiotic systems, the association between Crematogaster (Myrmicinae) and Macaranga (Euphorbiaceae) in South-East Asia, we conducted a phylogenetic analysis of the ant partners. For the phylogenetic analysis partial mitochondrial cytochrome oxidase I and II were sequenced and Maximum Parsimony analysis was performed. The analyzed Crematogaster of the subgenus Decacrema fell into three distinct clades which are also characterized by specific morphological and ecological traits (queen morphology, host-plants, and colony structure). Our results supported the validity of our currently used morphospecies concept for Peninsula Malaysia. However, on a wider geographic range (including North and North-East Borneo) some morphospecies turned out to be species complexes with genetically quite distinct taxa. Our phylogenetic analysis and host association studies do not indicate strict cocladogenesis between the subgenus Decacrema and their Macaranga host plants because multiple ant taxa occur on quite distinct host-plants belonging to different clades within in the genus Macaranga. These results support the view that host-shifting or host-expansion is common in the ants colonizing Macaranga. Additionally, the considerable geographic substructuring found in the phylogenetic trees of the ants suggests that allopatric speciation has also played a role in the diversification and the current distribution of the Decacrema ants. PMID- 12742750 TI - Molecular evolution of mammalian ribonucleases 1. AB - There have been many studies on the chemistry of mammalian pancreatic ribonucleases (ribonucleases 1), but the functional biology of this family of homologous proteins is still largely unknown. Many studies have been performed on the molecular evolution and properties of this enzyme from species belonging to a large number of mammalian taxa, including paralogous gene products resulting from recent gene duplications. Novel ribonuclease 1 sequences were determined for three rodent species (gundi, brush-tailed porcupine, and squirrel), rabbit, a fruit bat, elephant, and aardvark, and the new sequences were used for deriving most parsimonious networks of ribonucleases from different mammalian orders, including earlier determined nucleotide sequences and also a larger set of protein sequences. Weak support for interordinal relationships were obtained, except for an Afrotheria clade containing elephant and aardvark. Results of current analyses and also those obtained 20 years ago on amino acid sequences confirm conclusions derived recently from larger data sets of other molecules. Several examples of recent gene duplications in ribonucleases 1 are discussed, with respect to illustrate the concepts of orthology and paralogy. Previously evidence was presented for extensive parallelism between sequence regions with attached carbohydrate (about one quarter of the molecule) of unrelated species with cecal digestion (pig and guinea pig). These features are also present in the sequences of elephant and fruit bat, species with cecal digestion, but with a very low ribonuclease content in their pancreas. PMID- 12742751 TI - Evolutionary relationships among cyst-forming coccidia Sarcocystis spp. (Alveolata: Apicomplexa: Coccidea) in endemic African tree vipers and perspective for evolution of heteroxenous life cycle. AB - Cyst-forming coccidia of the genus Sarcocystis (Alveolata: Apicomplexa: Coccidea) parasitize vertebrates worldwide. Data from the small subunit rRNA genes (SSU) and the D2 domain of the large subunit rRNA genes were used to reconstruct phylogeny for all species in the Sarcocystidae for which sequences are currently available. We have focused on the evolutionary history of species that circulate between snakes as definitive hosts and rodents as intermediate hosts. Trees were reconstructed using maximum parsimony, minimum evolution, maximum likelihood and the bayesian phylogenetics. Our reconstructions support monophyly of Sarcocystidae but fail to robustly resolve the relationship within clades. Using a concatenated dataset of available rDNAs, the "isosporoid" coccidia Neospora, Toxoplasma, Besnoitia, Isospora and Hyaloklossia form a sister group to the monophyletic Sarcocystis. Moreover, we show that Sarcocystis from arboreal vipers of the genus Atheris, which are endemic to the mountain rain forests of the Equatorial Africa, are monophyletic, with sister species parasitizing the desert viper Pseudocerastes persicus from the Near East. We report the co-evolution of Sarcocystis spp. with their final snake hosts. The geological history of the African continent, mountain ranges, forests and general SSU rDNA rates were used to construct a linearized tree. Possible origin of the heteroxenous life cycle of Sarcocystis is discussed. PMID- 12742753 TI - Evolutionary relationships within the protostome phylum Sipuncula: a molecular analysis of ribosomal genes and histone H3 sequence data. AB - The phylogenetic relationships of the members of the phylum Sipuncula are investigated by means of DNA sequence data from three nuclear markers, two ribosomal genes (18S rRNA and the D3 expansion fragment of 28S rRNA), and one protein-coding gene, histone H3. Phylogenetic analysis via direct optimization of DNA sequence data using parsimony as optimality criterion is executed for 12 combinations of parameter sets accounting for different indel costs and transversion/transition cost ratios in a sensitivity analysis framework. Alternative outgroup analyses are also performed to test whether they affected rooting of the sipunculan topology. Nodal support is measured by parsimony jackknifing and Bremer support values. Results from the different partitions are highly congruent, and the combined analysis for the parameter set that minimizes overall incongruence supports monophyly of Sipuncula, but nonmonophyly of several higher taxa recognized for the phylum. Mostly responsible for this is the split of the family Sipunculidae in three main lineages, with the genus Sipunculus being the sister group to the remaining sipunculans, the genus Phascolopsis nesting within the Golfingiiformes, and the genus Siphonosoma being associated to the Phascolosomatidea. Other interesting results are the position of Phascolion within Golfingiidae and the position of Antillesoma within Aspidosiphonidae. These results are not affected by the loci selected or by the outgroup chosen. The position of Apionsoma is discussed, although more data would be needed to better ascertain its phylogenetic affinities. Monophyly of the genera with multiple representatives (Themiste, Aspidosiphon, and Phascolosoma) is well supported, but not the monophyly of the genera Nephasoma or Golfingia. Interesting phylogeographic questions arise from analysis of multiple representatives of a few species. PMID- 12742752 TI - Basal euteleostean relationships: a mitogenomic perspective on the phylogenetic reality of the "Protacanthopterygii". AB - Higher-level relationships of the basal Euteleostei (=Protacanthopterygii) are so complex and controversial that at least nine different morphology-based phylogenetic hypotheses have been proposed during the last 30 years. Relationships of the Protacanthopterygii were investigated using mitochondrial genomic (mitogenomic) data from 34 purposefully chosen species (data for 12 species being newly determined during the study) that fully represented major basal euteleostean lineages and some basal teleosts plus neoteleosts as outgroups. Unweighted and weighted maximum parsimony (MP) and maximum likelihood (ML) analyses were conducted with the data set that comprised concatenated nucleotide sequences from 12 protein-coding genes (excluding the ND6 gene and 3rd codon positions) and 22 transfer RNA (tRNA) genes (stem regions only) from the 34 species. The resultant trees were well resolved and largely congruent, with most internal branches being supported by high statistical values. Monophyly of the protacanthopterygians was confidently rejected by the mitogenomic data. Of the five major monophyletic groups that received high statistical support within the protacanthopterygians, a clade comprising members of the alepocephaloids was unexpectedly nested within the Otocephala, sister-group of the euteleosts. The remaining four major monophyletic groups, on the other hand, occupied phylogenetic positions intermediate between the otocephalans and neoteleosts, with a clade comprising esociforms + salmoniforms being more basal to the argentinoids and osmeroids. Although interrelationships of the latter two clades (argentinoids and osmeroids) with the neoteleosts remained ambiguous, the present results indicated explicitly that the protacanthopterygians as currently defined merely represent a collective, polyphyletic group of the basal euteleosts, located between the basal teleosts (elopomorphs and below) and neoteleosts (stomiiforms and above). PMID- 12742754 TI - Evolution of the prokaryotic protein translocation complex: a comparison of archaeal and bacterial versions of SecDF. AB - Protein translocation across the prokaryotic plasma membrane occurs at the translocon, an evolutionarily conserved membrane-embedded proteinaceous complex. Together with the core components SecYE, prokaryotic translocons also contain auxilliary proteins, such as SecDF. Alignment of bacterial and archaeal SecDF protein sequences reveals the presence of a similar number of homologous regions within each protein. Moreover, the conserved sequence domains in the archaeal proteins are located in similar positions as their bacterial counterparts. When these domains are, however, compared along Bacteria-Archaea lines, a much lower degree of similarity is detected. In Bacteria, SecDF are thought to modulate the membrane association of SecA, the ATPase that provides the driving force for bacterial protein secretion. As no archaeal version of SecA has been detected, the sequence differences reported here may reflect functional differences between bacterial and archaeal SecDF proteins, and by extension, between the bacterial and archaeal protein translocation processes. Moreover, the apparent absence of SecDF in several completed archaeal genomes suggests that differences may exist in the process of protein translocation within the archaeal domain itself. PMID- 12742755 TI - Cercopithecine Y-chromosome data provide a test of competing morphological evolutionary hypotheses. AB - We report here the results of the first molecular evolutionary analysis to include members of all 10 extant genera of cercopithecine monkeys. A total of 44 individuals were surveyed for approximately 2.2 kb of the testis-specific protein, Y-chromosome (TSPY). The TSPY sequences were subjected to parsimony analyses in PAUP 4.0, followed by tree comparison tests designed to assess existing morphological hypotheses of cercopithecine evolution. The results of these tests show that the present Y-chromosome dataset unambiguously supports: (1) monophyly of Macaca, (2) polyphyly of the mangabeys (Cercocebus and Lophocebus), (3) paraphyly of Cercopithecus, and (4) inclusion of Allenopithecus and Miopithecus in the tribe Cercopithecini. A number of unexpected Y-chromosome relationships are also discussed, including a pattern suggesting resurrection of the genus Chlorocebus for the guenons currently identified as Erythrocebus patas, Cercopithecus aethiops, and Cercopithecus lhoesti. Relative rate tests reveal significant difference in the TSPY substitution rate across numerous lineages in the tribe Cercopithecini. Because the rate differences follow no obvious phylogenetic pattern, "local" molecular clocks were not employed and divergence dates were not estimated for this tribe. In contrast, similar analysis of the Papionini reveals rate heterogeneity between a single pair of taxonomic groups: Macaca vs. the "African papionins." Divergence dates were therefore calculated for the tribe by calibrating TSPY clocks specific to each of these two clades. PMID- 12742756 TI - The misleading effects of composite taxa in supermatrices. AB - With the amount of available sequence data rapidly increasing, supermatrices are at the forefront of systematic studies. As an alternative to supertrees, supermatrices utilize a total evidence approach where different genes and other lines of data are merged into a single data matrix, which is then analyzed in an attempt to obtain the phylogeny that best explains the data. However, questions may arise when combining data sets in which one or more taxa do not have sequences available for each individual gene. Two possible solutions to this situation are to either leave all taxa separate and code unavailable sequences as missing, or to combine taxa at a level for which monophyly is assumed a priori. By reanalyzing the previous work of, we show that combining taxa may yield misleading results, i.e., hypotheses of relationships that are not supported by the underlying data. PMID- 12742757 TI - Assessing internal support with large phylogenetic DNA matrices. PMID- 12742758 TI - Codon bias variation in C-mos between squamate families might distort phylogenetic inferences. PMID- 12742759 TI - Contamination and chimerism are perpetuating the legend of the snake-eating cow with twisted horns (Pseudonovibos spiralis). A case study of the pitfalls of ancient DNA. PMID- 12742760 TI - Age-related differences in learning disabled and skilled readers' working memory. AB - This study determines whether age-related deficits in learning disabled (LD) readers' working memory performance reflect delays in retrieval efficiency and/or storage capacity. The study compared LD and skilled readers' working memory performance (N=226) across four age groups (7, 10, 13, and 20) for phonological, visual-spatial, and semantic information under initial (non-cued), gain (cues that bring performance to an asymptotic level), and maintenance conditions (asymptotic conditions without cues). The important results were that LD readers' working memory performance was inferior to skilled readers on verbal and visual spatial working memory tasks across all age groups and these differences increased on gain and maintenance conditions when compared to initial conditions. These reading group differences remained when age, reading, and mathematics were partialed from the analysis. The results support a general capacity explanation of reading group differences that is not totally dependent on reading skill. These differences in capacity reflect demands placed on both the accessing of new information and the maintenance of old information that extend beyond the phonological system. PMID- 12742761 TI - Validity of a test of children's suggestibility for predicting responses to two interview situations differing in their degree of suggestiveness. AB - In the present study the relative contributions of internal and external sources of variation in children's suggestibility in interrogative situations were examined. One hundred and eleven children (48 4- to 5-year-olds and 63 7- to 8 year-olds) were administered a suggestibility test (BTSS) and the most suggestible (N=36) and the least suggestible (N=36) children were randomly assigned to either an interview condition containing several suggestive techniques or to one containing only suggestive questions. The effects of internal sources of variation in suggestibility were compared with the effects of the interview styles on the children's answers. The former did influence the children, but the external sources of variation in suggestibility had a stronger impact. Influences of cognitive, developmental factors could be found, but not when abuse-related questions were asked and high pressured interview methods were used. These findings indicate that individual assessment of suggestibility can be of some assistance when interviewing children, but diminishing suggestive influences in interrogations must be given priority. PMID- 12742762 TI - What makes counting count? Verbal and visuo-spatial contributions to typical and atypical number development. AB - Williams Syndrome (WS) is marked by a relative strength in verbal cognition coupled with a serious impairment in non-verbal cognition. A strong deficit in numerical cognition has been anecdotally reported in this disorder; however, its nature has not been systematically investigated. Here, we tested 14 children with WS (mean age=7 years 2 months), 14 typically developing controls individually matched on visuo-spatial ability (mean age=3 years 5 months) as well as a larger group of typically developing controls (mean age=3 years 4 months) on two tasks to assess their understanding that counting determines the exact quantity of sets (cardinality principle). The understanding of the cardinality principle in children with WS is extremely delayed and only at the level predicted by their visuo-spatial MA. In this clinical group, only language accounted for a significant amount of the variance in cardinality understanding, whereas in the normal comparison group only visuo-spatial competence predicted the variance. The present findings suggest that visuo-spatial ability plays a greater role than language ability in the actual development of cardinality understanding in typically developing children, whereas the opposite obtains for the clinical group. PMID- 12742764 TI - An introduction to cancer communication and aging: theoretical and research insights. PMID- 12742763 TI - Speech timing and working memory in profoundly deaf children after cochlear implantation. AB - Thirty-seven profoundly deaf children between 8- and 9-years-old with cochlear implants and a comparison group of normal-hearing children were studied to measure speaking rates, digit spans, and speech timing during digit span recall. The deaf children displayed longer sentence durations and pauses during recall and shorter digit spans compared to the normal-hearing children. Articulation rates, measured from sentence durations, were strongly correlated with immediate memory span in both normal-hearing and deaf children, indicating that both slower subvocal rehearsal and scanning processes may be factors that contribute to the deaf children's shorter digit spans. These findings demonstrate that subvocal verbal rehearsal speed and memory scanning processes are not only dependent on chronological age as suggested in earlier research by. Instead, in this clinical population the absence of early auditory experience and phonological processing activities before implantation appears to produce measurable effects on the working memory processes that rely on verbal rehearsal and serial scanning of phonological information in short-term memory. PMID- 12742765 TI - Narratives and healing: exploring one family's stories of cancer survivorship. AB - This study investigates the narratives of one couple who lived through life changing events following a cancer diagnosis. The narratives of the cancer survivor and her husband are explored as they struggle to cope with their situation, provide support for one another, and consider their changing personal identities. This research addresses the communication dilemmas that often occur when family members, friends, and providers do not know how to respond to an individual diagnosed with cancer. The rationale for this study is threefold. First, this study advocates the need for learning about the composition of survivor identities over the course of a life-threatening illness. Second, this study seeks to understand how illness survivors and their family members use narratives as a method of communicating their changing identities. Finally, communicating about illness is often perceived as 'taboo', and this study may encourage others to be a part of the participants' stories and learn more about why those stories are often concealed. We learn from these three narratives that supportive relationships are central to healing and that it is through communication among family members that identities are composed and recomposed throughout the illness journey. This research affects communication, social support, identity, and emotion literature and is aligned with human appraisal theories as well. Finally, it offers insights into the ways in which we talk about, hear about, and learn about illness. PMID- 12742766 TI - Social identity and health: an intergroup communication approach to cancer. AB - This article describes the ways in which group identifications and stereotypes can inform our understanding of cancer prevention and treatment as well as more general social processes surrounding the experience of cancer. From a perspective grounded in social identity theory, we describe the ways in which understanding primary identities (i.e., those associated with large social collectives such as cultural groups), secondary identities (i.e., those associated with health behaviors), and tertiary identities (i.e., those associated with cancer) can help explain certain cancer-related social processes. We forward a series of propositions to stimulate further research on this topic. PMID- 12742767 TI - The impact of communication on cancer risk, incidence, morbidity, mortality, and quality of life. AB - Health communication has great potential to help reduce cancer risks, incidence, morbidity, and mortality while enhancing quality of life across the continuum of cancer care (prevention, detection, diagnosis, treatment, survivorship, and end of-life care). Effective health communication can encourage cancer prevention, inform cancer detection and diagnosis, guide cancer treatment, support successful cancer survivorship, and promote the best end-of-life care. This article examines the influences of health communication in confronting cancer and promoting important health outcomes. Implications of this analysis are drawn for directing informed cancer communication research and practice. PMID- 12742768 TI - Embedded health behaviors from adolescence to adulthood: the impact of tobacco. AB - Prevention of cancer risk behaviors before they become embedded in an individual's life is crucial. Health-related behaviors should be viewed for their embeddedness, critical aspects of which are (a) the complexity of the behavior itself; (.b) factors, both biological and psychological, within the individual communicator; (c) and external situational or sociocultural factors. The more extensively a behavior is embedded, the more difficult it will be to alter. Relative levels of embeddedness of the risk behavior and its entanglement with other nonrisky behaviors will evolve and change throughout one's life course. Smoking across the life span provides an excellent example of a thoroughly integrated, embedded behavior. How smoking is embedded with other behaviors changes from adolescence, where biological factors may be less salient and habit strength less pronounced, through adulthood, where habit strength is greater but health concerns are a more predictive factor. Researchers can produce more focused communication interventions by examining how health-endangering behaviors are embedded among benign behaviors or among other potentially dangerous behaviors. Ideally, the pattern of health behavior embeddedness should be analyzed prior to developing intervention communication strategies. PMID- 12742769 TI - Health, communication, and aging: cancer and older adults. AB - The diagnosis of cancer is devastating to the patient as well as to the formal and informal relationship support networks of the individual who must now cope with the disease. This article highlights the significant interpersonal communication and relationship issues that affect the overall quality of life for an older individual who has been diagnosed with cancer. Scholars who study the interpersonal behavior of older adults and those who study the consequences of a devastating diagnosis within the overall health communication context can make a significant contribution to our understanding of the process of cancer communication. Not only is the diagnosis of cancer life changing for the patient, but it also changes the dynamics of each relationship between the older adult who has cancer and those who will help to care for the cancer patient both in a formal institutional setting and at home. Our understanding of communication and aging for healthy as well as ill individuals can help us to better predict and explain the complexities of cancer communication. PMID- 12742770 TI - Cancer survivorship and agency model: implications for patient choice, decision making, and influence. AB - Relative to other types of health communication research (acute care physician patient communication, communication campaigns, compliance episodes, etc.), investigations of patient communication following the diagnosis of cancer are infrequent. Theoretically driven, empirical research is desperately needed in such postdiagnostic communication processes as survivorship, quality of life, palliative and hospice care, and loss, bereavement, and grief for those millions of people who have been diagnosed with the second leading cause of death in our nation. An organizational model of patient communication is needed that identifies and describes salient issues and processes involved when cancer patients attempt to negotiate the difficult courses of action following the diagnosis of cancer. The cancer survivorship and agency model (CSAM) proposes both general and specific strategies that serve as options for patients seeking to take greater control of the decision-making process related to their treatment and care of cancer. Although seemingly practical in its offering, CSAM is intended to serve as a heuristic springboard for theoretically based, applied communication research focusing exclusively on post diagnostic cancer processes. PMID- 12742771 TI - Assessing communication competence in an online study: toward informing subsequent interventions among older adults with cancer, their lay caregivers, and peers. AB - Employing quantitative and qualitative measures, online and paper versions, we tested Kreps's (1988) relational health communication model by examining relations among social support, communication competence, and perceived stress in a study of well-elders, elderly individuals with cancer, and their lay caregivers (N = 76). Grounding the qualitative part of the study in the narrative paradigm (Fisher, 1984), we used the critical incident technique (Flanagan, 1954) to collect participant narratives focusing on positive and negative expressions of social support. The results indicated partial support for the relational health communication model. In particular, participants who were more communicatively competent were found to have lower levels of perceived stress, and they were more satisfied with the support offered by members of their support network. In addition, the critical incidents revealed that participants received significantly more emotional and esteem support from their network than other social support types. Key limitations and future directions are also identified. PMID- 12742772 TI - The communication of palliative care for the elderly cancer patient. AB - Palliative care (PC) is often recommended by physicians for their elderly patients who are terminally ill. In contrast to hospice care, which precludes the use of any curative treatment at life's end stages, PC seeks primarily to comfort patients and to keep them pain free, yet it does not necessarily preclude medical treatment. It does seek to attend to patients' physical as well as psychological, emotional, spiritual, and existential needs in an attempt to enhance overall quality of life. A review of current literature in PC for oncology patients, elderly and otherwise, reveals a curious irony: Although PC plausibly entails a holistic, patient-centered approach to health care, much of the research on PC and, apparently, many of the practices in PC focus almost exclusively on the biomedical approach to patient care, particularly in regard to pain and symptom management. Furthermore, few methods in PC research incorporate patients' narratives and lived experiences in the final stages of their lives. We argue that a holistic, patient-centered approach must guide research in PC, including the treatment of elderly patients as "active interpreters, managers, and creators of the meaning of their health and illness" (Vanderford, Jenks, & Sharf, 1997, p.14) and of the meaning of their lives. PMID- 12742773 TI - Impersonal, interpersonal, and hyperpersonal social support: cancer and older adults. AB - Although cancer occurs throughout the life span, many of the most frequently occurring types of cancer increase as we grow older. In fact, only cardiovascular disease accounts for more deaths in adults 65 years of age and older. One of the ways that cancer patients cope or adapt to their illness is through socially supportive communicative interactions and relationships. Cutrona and Russell (1990) argued that social support is multidimensional and suggested that social support is most effective when the support needs of the individual are consistent with the type of social support being offered by the support provider. From the communicative perspective, the notion of optimal matching between the types of social support desired and the type of social support offered is extended to include the type of relationship between the communicants. In addition, it is argued that computer-mediated social support can be superior to face-to-face social support. This article attempts to identify some of the conditions under which this is true. PMID- 12742774 TI - The CAUSE model: a research-supported aid for physicians communicating with patients about cancer risk. AB - "Talk to your doctor" may be one of the most common pieces of advice given to help patients manage cancer risk. In fact, though, the support given to physicians for talking with patients about cancer prevention is not extensive. To address this need, we propose a decision aid for physicians. The CAUSE model draws on existing research, identifying common sources of tension or confusion in physician patient interaction. Specifically, the model assists physicians in anticipating common barriers to effective communication and overcoming them to establish credibility, create awareness and understanding, gain agreement about solutions, and help patients enact solutions for the prevention of cancer. PMID- 12742775 TI - Achieving the educational value of diversity. PMID- 12742776 TI - The need for diversity in higher education. AB - The author states that there is a need in all higher education, including the health professions, for racial, ethnic, and other kinds of diversity, and describes two court cases involving the University of Michigan's undergraduate and law school affirmative action admission policies. The outcomes of these cases will profoundly affect the quality of U.S. higher education, professional education, and graduate education. Affirmative action is one of the tools that many universities use to ensure the kind of comprehensively diverse student body that helps teach students to participate fully in this country's heterogeneous democracy and the global economy. Students are exposed to classmates with different life experiences, their prior assumptions are challenged, and they discover what they and their classmates have in common. And a variety of Fortune 500 corporations state that employees and managers who graduated from institutions with diverse student bodies demonstrate a variety of key skills that are crucial in the U.S. workplace. The author discusses why "colorblind" and "socioeconomically oriented" admission policies do not work and that they have only a tiny effect on white students' chances of acceptance. Until K-12 education is greatly improved for minorities, affirmative action is needed to give a "leg up" to students who might not otherwise be admitted but who can do the academic work. In medical education, there is a special urgency for diversity, since it is known that minority physicians are more likely to practice in areas where there are high concentrations of minorities. PMID- 12742777 TI - Blueprint for establishing an effective Postbaccalaureate medical school pre entry program for educationally disadvantaged students. AB - The purpose of this article is to provide public and private medical schools with a pragmatic blueprint for the development and implementation of an effective medical school pre-entry program that increases the pool of students interested in returning to health care shortage areas. An ancillary benefit of this program is an increase in the number of underrepresented minority students to medical schools. The structure, experiences, and results of the University of California, Davis, School of Medicine's Postbaccalaureate Reapplicant Program are used as a case study to construct the blueprint for returning 85-90% of program participants to shortage areas while increasing minority student admissions. The UC Davis program has been in place since 1991 and post-program acceptance rates have varied from 57% to 100% with an overall acceptance rate of 90.4% through 1999-00. Of 115 participating students who had previously been rejected by medical schools, 104 were accepted to health professional programs: 95 students were accepted to major U.S. medical schools and nine were accepted to masters in public health programs, physician's assistant programs, and one international medical school. This success rate has been achieved through a combination of intense assistance in study skills and test-taking skills, academic course work, and academic and pre-professional counseling. PMID- 12742778 TI - MEDPREP--30 years of making a difference. AB - The Medical/Dental Education Preparatory Program (MEDPREP) of Southern Illinois University School of Medicine has served over 1000 educationally or economically disadvantaged students since its inception in 1972. Seventy-eight percent of the former students (95% of the current students) are members of AAMC-defined underrepresented minority groups (URMs). Over the course of its 30-year history, 68% of the students have been accepted to professional schools, with 92% being accepted to medical schools and attending 90 different medical schools in the country. Of those accepted to professional schools, 87% have graduated or are expected to graduate. MEDPREP has had this impact on the profession by staying true to its original mission, assisting minority students and other students with disadvantaged backgrounds to prepare for admission and success in medical school. MEDPREP is an environment of high expectations, teamwork, personal growth, and opportunities to achieve goals. Students are served by an advisor who helps structure an individualized curriculum designed to develop a knowledge base and learning and reasoning skills. The success of the program is due to the school's commitment and support, the faculty who serve the students, and the sacrifice and hard work of the MEDPREP students. PMID- 12742779 TI - Increasing access to medical education for students from medically underserved communities: one program's success. AB - The Premedical Honors College (PHC) is an eight-year, BS-MD program created in 1994 by Baylor College of Medicine (BCM) and The University of Texas-Pan American (UT-PA) to increase the number of physicians addressing the health care needs of underserved populations in Texas. The PHC targets South Texas, a 13-county, medically underserved area with a population that is 82% Hispanic. To date, the PHC has had 159 matriculants and 71 graduates, of whom 60 (84.5%) have matriculated into medical school. These results are significant considering that in 1996, only four students from all five South Texas colleges (combined enrollment of 30000 students) were accepted to medical school. An outcomes study comparing PHC matriculants with students of similar academic ability, ethnicity, and interest in medicine revealed that the odds of medical school matriculation were seven times higher for PHC students than for non-PHC students. The PHC's initial success has been acknowledged by the Texas legislature, which recently passed a bill to promote the PHC's replication. In addition, the number of PHC students-of whom 95% are Mexican American-who matriculate into medical school annually is significant nationally. In 2001, only 386 U.S. medical school matriculants (2.3% of all matriculants) were Mexican American; 17 of these students (4.4%) were PHC graduates. If current trends continue, the PHC could significantly expand the number of physicians serving minority and medically underserved populations in Texas and the nation. Also, the PHC provides an opportunity for research on programs designed to create pathways from high school to medical school. PMID- 12742780 TI - Educational benefits of diversity in medical school: a survey of students. AB - PURPOSE: Many U.S. medical schools have abandoned affirmative action, limiting the recruitment and reducing the admission of underrepresented minority (URM) students even though research supports the premise that the public benefits from an increase in URM physicians and that URM physicians are likely to serve minority, poor, and Medicaid populations. Faculty and students commonly assume they benefit from peer cultural exchange, and the published evidence for the past two decades supports this notion. This research examined the students' perceptions of the educational merits of a diverse student body by surveying medical students at two schools. METHOD: In 2000, medical students from all four years at Harvard Medical School and the University of California, San Francisco, School of Medicine were enrolled in a telephone survey about the relevance of racial diversity (among students) in their medical education. Students responded to the interviewer's questions on a five-point Likert-type scale. RESULTS: Of the 55% of students who could be located, 97% responded to the survey. Students reported having little intercultural contact during their formative years but significantly more interactions during higher education years, especially in medical school. Students reported contacts with diverse peers greatly enhanced their educational experience. They strongly supported strengthening or maintaining current affirmative action policies in admissions. The responses and demography of the Harvard and UCSF students did not differ significantly, nor did they differ for majority students and URM students-all groups overwhelmingly thought that racial and ethnic diversity among their peers enhanced their education. CONCLUSIONS: Diversity in the student body enhanced the educational experiences of students in two U.S. medical schools. PMID- 12742781 TI - Characteristics of health professions schools, public school systems, and community-based organizations in successful partnerships to increase the numbers of underrepresented minority students entering health professions education. AB - PURPOSE: To identify characteristics of health professions schools, public schools, and community-based organizations in successful partnerships to increase the number of underrepresented minority students entering health professions. The Robert Wood Johnson Foundation and the W. K. Kellogg Foundation funded the Health Professions Partnership Initiative program developed from Project 3000 by 2000 of the Association of American Medical Colleges. METHOD: Semi-structured interviews were completed with awardees and representatives of the funding agencies, the national program office, and the national advisory committee between the fall of 2000 and the summer of 2002. Site visits were conducted at ten sites, with representatives of partner institutions, teachers, parents, and children. Characteristics that supported and hindered development of successful partnerships were identified using an iterative qualitative approach. RESULTS: Successful partnerships included professional schools that had a commitment to community service. Successful leaders could work in both cultures of the professional and public schools. Attitudes of respect and listening to the needs of partners were essential. Public school governance supported innovation. Happenstance and convergence of interests played significant roles in partnership development. The most telling statement was "We did it, together." CONCLUSIONS: This study identifies characteristics associated with smoothly working partnerships, and barriers to successful program development. Successful partnerships can form the basis on which educational interventions are built. The study is limited by the definition of success used, and its focus on one funded program. The authors were unable to identify outcomes in terms of numbers of children influenced by programs or instances in which lasting changes in health professions schools had occurred. PMID- 12742783 TI - USMLE performances in a predominantly Asian and Pacific Islander population of medical students in a problem-based learning curriculum. AB - PURPOSE: To compare the USMLE performances of students of various ethnicities, predominantly Pacific Islander and Asian, at one medical school and to examine the predictive validity of MCAT scores for USMLE performance. METHOD: A total of 258 students in the graduating classes of 1996-2000 at the University of Hawai'i School of Medicine were classified by ethnicity. Demographic and performance characteristics of the groups were examined, and MCAT scores with and without undergraduate science GPA were used to predict USMLE performance. Under- and over prediction rates were computed for each ethnic group. RESULTS: Ethnic groups did not differ significantly by gender or undergraduate GPA. Chinese, Caucasian, and Other Asian students tended to have higher MCAT scores than Hawaiian/other Pacific Islander, and Filipino students. Ethnic groups did not differ significantly in prediction of USMLE Step 1 performance. For Step 2, MCAT scores significantly over-predicted performance of Filipino students and tended to under predict performance of Caucasian students. CONCLUSION: Although MCAT scores and science GPA were good predictors of USMLE performance, ethnic differences were found in the degrees of their predictive validity. These findings both replicate and extend results of earlier studies, and again point to the importance of exploring additional predictor variables. The authors encourage future research on the effects of the following factors on success in medical school: reading and test-taking skills, socio-cultural and environmental influences on learning, communication styles, primary language use, family support, and family responsibilities. PMID- 12742784 TI - From case-based reasoning to problem-based learning. AB - Many medical schools today are questioning whether they should implement a problem-based learning (PBL) curriculum. Educators have raised serious questions regarding the efficiency of PBL and therefore recommend learning more about the cognitive processes developed by PBL before implementing it broadly. In addition, it is important to determine whether PBL best matches the human reasoning process. The authors' theoretical discussion examines the relationship between the case-based reasoning (CBR) model and the PBL model. CBR indicates that the knowledge source one uses while solving a new problem includes not only generalized rules or general cases, but often a memory of stored cases recording specific prior episodes. CBR enables the reasoner to recommend solutions to problems quickly and to propose solutions in domains that are not completely understood, such as medicine. The authors' analysis reveals a strong association between the CBR and PBL models, and thus it can be argued that PBL is a successful teaching method that should be encouraged by medical schools. PMID- 12742786 TI - Still life with open window. PMID- 12742788 TI - Gender differences in academic advancement: patterns, causes, and potential solutions in one US College of Medicine. AB - PURPOSE: The influx of women into academic medicine has not been accompanied by equality for male and female faculty. Women earn less than men in comparable positions, progress more slowly through academic ranks, and have not attained important leadership roles. This study tested hypotheses about why gender disparities exist in salary, rank, track, leadership, and perceptions of campus climate at one academic center, the University of Arizona College of Medicine, Tucson. METHOD: Salary, rank, and track data were obtained from institutional databases for the 1999-2000 fiscal year. A structured, online questionnaire was made available to 418 faculty members to collect information about their goals, attitudes, and experiences. RESULTS: A total of 198 faculty members completed the questionnaire. The data showed significant gender differences in faculty salaries, ranks, tracks, leadership positions, resources, and perceptions of academic climate. On average, women earned US dollars 12777 or 11% less than men, after adjusting for rank, track, degree, specialty, years in rank, and administrative positions (p <.0003). Of female faculty, 62% were assistant professors (49% of women were non-tenure-eligible assistant professors), while 55% of male faculty were promoted and tenured. Almost a third of women reported being discriminated against, compared with only 5% of men (p <.00001). CONCLUSION: Substantial gender differences in the rewards and opportunities of academic medicine remain, that can not be attributed to differences in productivity or commitment between women and men. PMID- 12742789 TI - Assessing medical students' clinical sciences knowledge in France: a collaboration between the NBME and a consortium of French medical schools. AB - PURPOSE: The French government, as part of medical education reforms, has affirmed that an examination program for national residency selection will be implemented by 2004. The purpose of this study was to develop a French multiple choice (MC) examination using the National Board of Medical Examiners' (NBME) expertise and materials. METHOD: The Evaluation Standardisee du Second Cycle (ESSC), a four-hour clinical sciences examination, was administered in January 2002 to 285 medical students at four university test sites in France. The ESSC had 200 translated and adapted MC items selected from the Comprehensive Clinical Sciences Examination (CCSE), an NBME subject test. RESULTS: Less than 10% of the ESSC items were rejected as inappropriate to French practice. Also, the distributions of ESSC item characteristics were similar to those reported with the CCSE. The ESSC also appeared to be very well targeted to examinees' proficiencies and yielded a reliability coefficient of.91. However, because of a higher word count, the ESSC did show evidence of speededness. Regarding overall performance, the mean proficiency estimate for French examinees was about 0.4 SD below that of a CCSE population. CONCLUSIONS: This study provides strong evidence for the usefulness of the model adopted in this first collaborative effort between the NBME and a consortium of French medical schools. Overall, the performance of French students was comparable to that of CCSE students, which was encouraging given the differences in motivation and the speeded nature of the French test. A second phase with the participation of larger numbers of French medical schools and students is being planned. PMID- 12742790 TI - ASSERT: the effectiveness of a continuing medical education video on knowledge and attitudes about interpersonal violence. AB - PURPOSE: Developing ways to educate busy clinicians is especially challenging when the subject includes medical, social, and legal aspects, as is the case with interpersonal violence (IPV). Organizations such as the American Medical Association and the Joint Commission on Accreditation of Healthcare Organizations (JCAHO) recommend routine IPV screening for patients. Videotape efficiently provides training in multiple locations using experts from different fields. The authors created and evaluated a multidisciplinary continuing medical education (CME) videotape on IPV. METHOD: The video, ASSERT: A Guide to Child, Elder, Sexual, and Domestic Abuse for Medical Professionals, was developed by experts from medicine, social work, nursing, and law. The video featured role-plays to demonstrate different approaches to these difficult clinical encounters. Pre- and post-viewing questionnaires assessed the video's effectiveness. RESULTS: In all, 120 physicians and 172 other personnel (e.g., nurses, social workers) at 24 sites associated with four academic medical centers completed paired questionnaires. Using a conservative level of significance (p <.002), there was significant improvement for physicians in 77% of the knowledge items and 75% of the attitude items from pre- to post-viewing questionnaires. A total of 73% of viewers would recommend the video to colleagues. CONCLUSIONS: The IPV video, using experts from multiple disciplines, improved knowledge and attitudes about child, elder, sexual, and domestic violence, and was rated highly by clinicians. The video was useful for preparing for a JCAHO accreditation visit. PMID- 12742791 TI - Can internal medicine residents master microscopic urinalysis? Results of an evaluation and teaching intervention. AB - PURPOSE: Although microscopic urinalysis (micro UA) is commonly used in clinical practice, and residents are trained in micro UA, proficiency in this procedure has not been studied. METHOD: In 1996-97, 38 residents in the University of Nebraska Medical Center's internal medicine (IM) residency program were evaluated on their technical ability to perform micro UA, and on their cognitive skills in recognizing common micro UA findings. After identifying deficits in the residents' cognitive competency, two educational interventions were applied and residents were tested after each intervention. RESULTS: A total of 24 residents (63%) correctly prepared the specimen for analysis (the technical portion). On the cognitive portion, only one of the 38 residents correctly identified 80% of all micro UA findings in the urinary sediment, although 11 (29%) residents identified UA findings specific to urinary tract infection (UTI). The first educational intervention did little to improve residents' performance. A second more intensive intervention resulted in 10 (45%) residents identifying 80% of all micro UA findings, and 19 (86%) residents correctly identifying UTI findings. CONCLUSIONS: Many residents were not proficient in performing micro UA, even after intensive educational interventions. Although micro UA is a simple procedure, residents' mastery cannot be assumed. Residency programs should assess competency in this procedure. PMID- 12742792 TI - Assessing medical students' training in end-of-life communication: a survey of interns at one urban teaching hospital. AB - PURPOSE: Although interns are responsible for caring for dying patients, little is known about end-of-life education and training, including communication skills, in U.S. medical schools. This study of three consecutive cohorts of new interns assessed their perceptions of the amount and types of classroom and clinical instructional strategies used during medical school, their self-rated skill and comfort levels in different aspects of end-of-life communication, and the associations between these measures. METHOD: A self-administered questionnaire was given to three consecutive cohorts (1996-1998) of incoming interns (n = 162). Measures were self-reported amount and type of education and clinical experience with four end-of-life communication domains (giving bad news, discussing advance directives, discussing prognosis with the patient, and discussing with the patient's family) and self-perceived comfort and skill levels in relation to different types of end-of-life communication. RESULTS: A total of 157 interns completed the questionnaire. They reported very little classroom teaching, clinical observation, or clinical experience with end-of-life communication during medical school. They lacked comfort and skill in the end-of life communication domains that were studied. More reported clinical observation and experience with caring for and communicating with dying patients was associated with greater perceived comfort and skill, while classroom teaching was not. CONCLUSIONS: These interns, mostly U.S. medical school graduates (98.7%, n = 155) reported little training and low self-perceived comfort and skill with important elements of end-of-life communication that might contribute to a lack of preparedness to address these issues during their internship. Further research that confirms and explains the underlying reasons for these findings seems warranted. PMID- 12742794 TI - Is virtual the same as real? Medical students' experiences of a virtual patient. AB - PURPOSE: Narrative and problem-solving versions of the same virtual patient's case were created for teaching communication skills to medical students. This qualitative study explored how students experienced the virtual patient. METHOD: In 1998-1999 in-depth, free-form interviews and follow-ups were conducted with 12 third-year medical students at Monash University in Australia. Students were asked about their experiences with the virtual patient. The interviews were qualitatively analyzed using psychological phenomenology. RESULTS: Results were in the form of a description of the students' lived experiences with the virtual patient. Findings indicated that students responded to the virtual patient as if she were real but they felt a simultaneous sense of prefabrication, which often led to frustration. Students' experiences of both versions were similar, but the narrative version permitted better rapport with the virtual patient. CONCLUSION: This phenomenological study indicated that a constructed, computer-based virtual patient can have substantial emotional effects on medical students. PMID- 12742795 TI - Value of humanitarian assistance missions in residency. AB - The educational value of participation in humanitarian assistance missions by third-year residents was assessed. PMID- 12742796 TI - Fever of unknown origin: the evolving definition. PMID- 12742797 TI - Headache and stroke: two common disorders or commonality of cause? PMID- 12742798 TI - The growing burden of tuberculosis: global trends and interactions with the HIV epidemic. AB - BACKGROUND: The increasing global burden of tuberculosis (TB) is linked to human immunodeficiency virus (HIV) infection. METHODS: We reviewed data from notifications of TB cases, cohort treatment outcomes, surveys of Mycobacterium tuberculosis infection, and HIV prevalence in patients with TB and other subgroups. Information was collated from published literature and databases held by the World Health Organization (WHO), the Joint United Nations Programme on HIV/Acquired Immunodeficiency Syndrome (UNAIDS), the US Census Bureau, and the US Centers for Disease Control and Prevention. RESULTS: There were an estimated 8.3 million (5th-95th centiles, 7.3-9.2 million) new TB cases in 2000 (137/100,000 population; range, 121/100,000-151/100,000). Tuberculosis incidence rates were highest in the WHO African Region (290/100,000 per year; range, 265/100,000 331/100,000), as was the annual rate of increase in the number of cases (6%). Nine percent (7%-12%) of all new TB cases in adults (aged 15-49 years) were attributable to HIV infection, but the proportion was much greater in the WHO African Region (31%) and some industrialized countries, notably the United States (26%). There were an estimated 1.8 million (5th-95th centiles, 1.6-2.2 million) deaths from TB, of which 12% (226 000) were attributable to HIV. Tuberculosis was the cause of 11% of all adult AIDS deaths. The prevalence of M tuberculosis-HIV coinfection in adults was 0.36% (11 million people). Coinfection prevalence rates equaled or exceeded 5% in 8 African countries. In South Africa alone there were 2 million coinfected adults. CONCLUSIONS: The HIV pandemic presents a massive challenge to global TB control. The prevention of HIV and TB, the extension of WHO DOTS programs, and a focused effort to control HIV-related TB in areas of high HIV prevalence are matters of great urgency. PMID- 12742799 TI - Combination therapy with angiotensin-converting enzyme inhibitors and angiotensin receptor antagonists in the treatment of patients with type 2 diabetes mellitus. AB - Aggressive therapy for patients with type 2 diabetes mellitus and renal disease is warranted given the natural history of this disease. Although antagonizing the renin-angiotensin system is clearly important, how this is accomplished is of considerable controversy. On the one hand, recent clinical trials of patients with type 2 diabetes mellitus with renal disease demonstrate unequivocally the renal protective effect of angiotensin receptor blockers (ARBs). Although the results of the recently published LIFE trial are encouraging, inconsistencies have been observed with ARBs in reducing cardiovascular end points. On the other hand, angiotensin-converting enzyme inhibitors have a dramatic effect in reducing cardiovascular events but have not been shown convincingly to reduce progression of renal disease in patients with type 2 diabetes mellitus. These studies leave us in a quandary as to the optimal initial treatment regimen for patients with type 2 diabetes mellitus and renal disease despite the recent recommendations from the American Diabetes Association (Alexandria, Va). Given the fact that many of these individuals will require administration of multiple antihypertensive agents, perhaps the initial treatment with a combination of an ARB and angiotensin-converting enzyme inhibitor affords optimal cardiovascular and renal protection for patients with type 2 diabetes mellitus and renal disease. Future clinical trials should be designed to address this issue. PMID- 12742800 TI - From prolonged febrile illness to fever of unknown origin: the challenge continues. AB - BACKGROUND: Epidemiological changes and the ongoing expansion of the diagnostic armamentarium warrant a regular update of the spectrum of diseases that present as prolonged febrile illnesses. METHODS: We prospectively collected a series of 290 immunocompetent patients referred to our university hospital between 1990 and 1999 with a febrile illness (temperature >38.3 degrees C) of uncertain cause, lasting at least 3 weeks. Patients were categorized in 4 groups according to the timing of diagnosis: early diagnosis (within 3 in-hospital days or 3 outpatient visits), intermediate diagnosis (between days 4 and 7), late diagnosis (after day 7), and no diagnosis during index contact or follow-up. RESULTS: A final diagnosis was established early in 67 patients (23.1%), intermediate in 38 (13.1%), and late in 87 (30.0%). In the remaining 98 (33.8%), no diagnosis was made. The cause of the fever remained obscure in 50 (47.6%) of 105 patients with episodic fever vs 48 (25.9%) of 185 patients with continuous fever (P<.001). Among the 192 patients with a final diagnosis, noninfectious inflammatory diseases represented the most prevalent diagnostic category (35.4%), surpassing infections (29.7%), miscellaneous causes (19.8%), and malignancies (15.1%). Fourteen disorders accounted for over 59% of diagnoses, whether diagnosis was reached early, intermediate, or late. Hematological malignancies made up 11.5% of diagnoses, but were responsible for 14 (58.3%) of the 24 fatalities related to the febrile illness. Of the 80 patients discharged alive without diagnosis and for whom follow-up was available, 3 died, but the deaths were considered to be unrelated to the feverish illness. CONCLUSIONS: Prolonged febrile illnesses remain a diagnostic challenge. Despite the technological progress of the late 20th century, the origin of the fever remains elusive in many patients, especially in those with episodic fevers. Noninfectious inflammatory diseases emerge as the most prevalent diagnostic category. PMID- 12742801 TI - National trends in antiobesity medication use. AB - BACKGROUND: The use of medications to treat obesity remains controversial. Our goal was to assess national trends in antiobesity medication use with a focus on patterns surrounding the 1997 removal of antiobesity drugs from the market. METHODS: Using a serial cross-sectional study design, we analyzed a nationally representative sample of US office-based physician visits from 1991 to 2002. Data come from the IMS HEALTH National Disease and Therapeutic Index. These data include a sample of 13 452 patient visits for which a diagnosis of clinical obesity was made, with annual visits ranging from 666 in 1994 to 1854 in 1996. The unit of analysis is the patient visit, while the primary outcome measures are the annual and quarterly number of antiobesity drug mentions for clinically obese patients. RESULTS: At its peak in the second quarter of 1997, 2.5 million Americans were taking antiobesity medications, a 4-fold increase over the prior 2 years. Although antiobesity medication use diminished following the market exit of fenfluramine hydrochloride and dexfenfluramine hydrochloride, current levels of use remain above those in the early 1990s. Phentermine has consistently been the most common antiobesity medication. In 2002, an annualized 1.2 million mentions of phentermine use were noted (31% of drug-treated obese patients). Newly released medications, orlistat (0.6 million) and sibutramine hydrochloride (0.4 million), were used less often. Most antiobesity medication use occurs in patients without other reported medical conditions. CONCLUSIONS: Use of antiobesity medications increased rapidly with public and professional interest in fenfluramine-phentermine (fen-phen) combination therapy. Despite reports of adverse outcomes associated with fenfluramine agents (fen-phen and dexfenfluramine), the use of these medication therapies did not diminish until soon before their removal from the market in 1997. PMID- 12742802 TI - Plasma total cholesterol level as a risk factor for Alzheimer disease: the Framingham Study. AB - BACKGROUND: Previous studies examining the association of plasma cholesterol levels with the risk for development of Alzheimer disease (AD) have been inconclusive. We examined the impact of baseline and lifetime plasma total cholesterol levels averaged across many years on the risk for AD in a large, population-based cohort. METHODS: Five thousand two hundred nine subjects from the Framingham Study original cohort underwent biennial evaluation for cardiovascular risk factors since 1950, with estimations of serum total cholesterol levels at 19 of these 25 biennial examinations. The study sample consisted of 1026 subjects from this cohort who were alive and free of stroke and dementia at examination cycle 20 (1988-1989) and had undergone apolipoprotein E (APOE) genotyping. The main outcome measure was incident AD diagnosed using standard criteria, according to average total cholesterol levels across biennial examination cycles 1 to 15 and baseline total cholesterol level measured at the 20th biennial examination cycle. RESULTS: Alzheimer disease developed in 77 subjects from 1992 to 2000. After adjustment for age, sex, APOE genotype, smoking, body mass index (calculated as weight in kilograms divided by the square of height in meters), coronary heart disease, and diabetes, we found no significant association between the risk for incident AD and average cholesterol level at biennial examination cycles 1 to 15 (hazard ratio per 10-mg/dL [0.3 mmol/L] rise, 0.95; 95% confidence interval, 0.87-1.04) or baseline total cholesterol level at examination 20 (hazard ratio, 0.97; 95% confidence interval, 0.90-1.05). CONCLUSION: In this large, population-based cohort, baseline and long term average serum total cholesterol levels were not associated with the risk for incident AD. PMID- 12742803 TI - Headache and the risk of stroke: a prospective observational cohort study among 35,056 Finnish men and women. AB - BACKGROUND: Previous studies have shown an increased risk of stroke among patients with migraine. However, very few data are available on the possible association between chronic unspecified headache and the risk of stroke. METHODS: A prospective cohort study including 35,056 randomly selected Finnish men and women aged 25 to 64 years at baseline who participated in a cardiovascular risk factor survey in 1972, 1977, 1982, or 1987. Self-reported headache, smoking, diabetes, blood pressure, weight, height, serum cholesterol level, and oral contraceptive use were recorded at baseline. During the follow-up, 2167 incident stroke events were ascertained with computer-based record linkage. RESULTS: Women reported headache twice as often as men (16.7% vs 8.9%). Among men, the headache associated hazard ratios (95% confidence intervals) for stroke were 4.08 (2.10 7.93), 1.86 (1.33-2.59), and 1.24 (1.05-1.47) during 1, 5, and a maximum of 23 years of follow-up, respectively. Adjustment for the other risk factors decreased the hazard ratios only slightly. Among women, there was also a direct but statistically nonsignificant association between headache and the risk of stroke. CONCLUSIONS: Chronic headache is an independent predictor of stroke among men. Since the association between headache and the risk of stroke was particularly strong during a short follow-up, chronic headache may be a marker of the underlying disease process leading to acute stroke. The sex difference observed in this association may be due to a higher prevalence and a more heterogeneous etiology of headache in women compared with men. PMID- 12742804 TI - Use of medications and dietary supplements in later years among male former top level athletes. AB - BACKGROUND: The association between sports participation and later need of medications and dietary supplements is unknown. SUBJECTS AND METHODS: Male athletes (N = 2026) who had represented Finland in international events from 1920 through 1965 and 1401 control subjects who had been classified healthy at the age of 20 years participated in this population-based cohort study. MAIN OUTCOME MEASURES: The main outcome measures were reimbursable medications for hypertension, cardiac insufficiency, coronary heart disease, diabetes, and asthma identified from the national registry from 1970 through 1998 as well as the use of nonsteroidal anti-inflammatory drugs, antacids, and specific vitamin and mineral supplements for at least 60 days during the past year reported by questionnaire in 1985. RESULTS: Among former top-level athletes compared with controls, the probability of initiating medication was decreased for cardiac insufficiency (age-adjusted hazard ratio [HR], 0.61; 95% confidence interval [CI], 0.50-0.74; P<.001), coronary heart disease (age-adjusted HR, 0.72; 95% CI, 0.58-0.89; P=.002), and asthma (age-adjusted HR, 0.47; 95% CI, 0.36-0.66; P<.001). Furthermore, the risk of initiation of treatment with regular medication for hypertension (age-adjusted HR, 0.73; 95% CI, 0.54-1.00; P=.046) and diabetes (age-adjusted HR, 0.38; 95% CI, 0.20-0.73; P=.004) was reduced for endurance athletes but not for power athletes. In 1985, compared with control subjects, athletes used fewer nonsteroidal anti-inflammatory drugs (age-adjusted odds ratio [OR], 0.48; 95% CI, 0.35-0.67; P<.001) and antacids (age-adjusted OR, 0.49; 95% CI, 0.31-0.77; P=.002) but more vitamin A (age-adjusted OR, 1.87; 95% CI, 1.24 2.82; P=.003), vitamin B (age-adjusted OR, 2.26; 95% CI, 1.64-3.12, P<.001), vitamin C (age-adjusted OR, 1.96; 95% CI, 1.45-2.63; P<.001), selenium (age adjusted OR, 1.62; 95% CI, 1.15-2.28; P=.006), and iron (age-adjusted OR, 2.35; 95% CI, 1.33-4.15; P=.003) supplements. CONCLUSION: The need for long-term therapy for cardiac disease and asthma as well as for treatment with nonsteroidal anti-inflammatory drugs and antacids is reduced among former top-level athletes, but the use of dietary supplements is increased. PMID- 12742805 TI - Effects of blood pressure lowering with perindopril and indapamide therapy on dementia and cognitive decline in patients with cerebrovascular disease. AB - BACKGROUND: High blood pressure and stroke are associated with increased risks of dementia and cognitive impairment. This study aimed to determine whether blood pressure lowering would reduce the risks of dementia and cognitive decline among individuals with cerebrovascular disease. METHODS: The Perindopril Protection Against Recurrent Stroke Study (PROGRESS) was a randomized, double-blind, placebo controlled trial conducted among 6105 people with prior stroke or transient ischemic attack. Participants were assigned to either active treatment (perindopril for all participants and indapamide for those with neither an indication for nor a contraindication to a diuretic) or matching placebo(s). The primary outcomes for these analyses were dementia (using DSM-IV criteria) and cognitive decline (a decline of 3 or more points in the Mini-Mental State Examination score). RESULTS: During a mean follow-up of 3.9 years, dementia was documented in 193 (6.3%) of the 3051 randomized participants in the actively treated group and 217 (7.1%) of the 3054 randomized participants in the placebo group (relative risk reduction, 12% [95% confidence interval, -8% to 28%]; P =.2). Cognitive decline occurred in 9.1% of the actively treated group and 11.0% of the placebo group (risk reduction, 19% [95% confidence interval, 4% to 32%]; P =.01). The risks of the composite outcomes of dementia with recurrent stroke and of cognitive decline with recurrent stroke were reduced by 34% (95% confidence interval, 3% to 55%) (P =.03) and 45% (95% confidence interval, 21% to 61%) (P<.001), respectively, with no clear effect on either dementia or cognitive decline in the absence of recurrent stroke. CONCLUSIONS: Active treatment was associated with reduced risks of dementia and cognitive decline associated with recurrent stroke. These findings further support the recommendation that blood pressure lowering with perindopril and indapamide therapy be considered for all patients with cerebrovascular disease. PMID- 12742807 TI - Prevalence and structure of palliative care services in California hospitals. AB - BACKGROUND: Most Americans die in hospitals where shortcomings in end-of-life care are endemic. Hospital-based palliative care services can improve the care of these patients, yet there are limited data regarding the availability of such services. We sought to determine the prevalence of palliative care services in California hospitals. METHODS: We conducted a cross-sectional survey of a random sample of 25% of all California hospitals. We recorded the percentage of hospitals reporting current or planned palliative care consultation services or inpatient palliative care units. RESULTS: We collected data from 107 (96%) of 112 hospitals. Only 17% of hospitals have a palliative care consult service, and 6% have an inpatient palliative care unit. Nearly all services are multidisciplinary. Twenty percent of hospitals have a contract to provide inpatient hospice beds, 19% have an outpatient-based hospice service affiliated with the hospital, and 74% offer bereavement services. Half of all palliative care services are funded exclusively by the hospital. Thirty-eight hospitals (36%) reported an interest in developing palliative care services. CONCLUSIONS: Few California hospitals currently have palliative care services, though more express interest in developing them. Bereavement and hospice services are more common and offer opportunities for increasing the number of palliative care services in hospitals. Further studies are needed to characterize palliative care services more fully and to assess the quality of care provided by these services. PMID- 12742806 TI - Relation of triglyceride levels, fasting and nonfasting, to fatal and nonfatal coronary heart disease. AB - BACKGROUND: It remains unclear whether hypertriglyceridemia is an independent risk factor for coronary heart disease (CHD), and whether fasting and nonfasting triglyceride (TG) levels are equally predictive. METHODS: A total of 2809 (of 12 866) men randomized during 1973 through 1975 into the Multiple Risk Factor Intervention Trial with fasting and nonfasting TG levels measured at baseline were followed up for CHD incidence and death. Proportional hazards regression models were used to assess associations of fasting and nonfasting TG levels with CHD. RESULTS: Average fasting and nonfasting TG levels were 187 and 284 mg/dL (2.11 and 3.21 mmol/L), respectively. Prevalence of hypertriglyceridemia (200 mg/dL [2.26 mmol/L] or more) was 31% for fasting and 61% for nonfasting. There were 175 nonfatal or fatal CHD events during 8 years and 328 CHD deaths during 25 years. Compared with TG levels less than 200 mg/dL, risk factor-adjusted hazard ratios for CHD mortality for hypertriglyceridemia were 1.24 (P =.09) for fasting and 1.26 (P =.07) for nonfasting. For nonfatal or fatal CHD, fasting and nonfasting TG levels were similarly predictive with hazard ratios of 1.64 (P =.004) for fasting and 1.46 (P =.03) for nonfasting. These associations for fasting TG levels were assessed to be underestimated by 56% because of regression dilution bias, with attenuation likely greater for nonfasting TG levels. CONCLUSIONS: Greater ease of obtaining nonfasting than fasting measurements, greater prevalence of hypertriglyceridemia with nonfasting than fasting values, and similarly increased risk with each indicate that nonfasting TG levels may be more useful than fasting ones for risk stratification. PMID- 12742808 TI - Influenza vaccination in community-dwelling elderly: impact on mortality and influenza-associated morbidity. AB - BACKGROUND: Influenza-related morbidity and mortality have been extensively studied with hospital and reimbursement data. However, little is known about the effectiveness of the annual vaccination programs in generally healthy community dwelling elderly. The objective of our study was to investigate the effectiveness of influenza vaccination in community-dwelling elderly during the 1996 to 1997 influenza epidemic. METHODS: We performed a population-based cohort study using the computerized Integrated Primary Care Information database in the Netherlands. Subjects who were 65 years and older in 1996 with a permanent status in a practice in the source population were considered eligible for study participation. Two cohorts were defined on the basis of vaccination status. We estimated and compared all-cause mortality, pneumonia, and clinical influenza infection rates between the cohorts. RESULTS: Influenza vaccination was associated with a significant reduction of morbidity and mortality in vaccinated elderly (relative risk [RR], 0.72; 95% confidence interval [CI], 0.60-0.87). Influenza infections decreased significantly in the vaccinated population (RR, 0.48; 95% CI, 0.26-0.91). Mortality was reduced significantly in elderly with comorbidity (RR, 0.67; 95% CI, 0.48-0.94). The risk reduction for pneumonia was nonsignificant (RR, 0.77; 95% CI, 0.55-1.07) but was temporally related to the peak influenza activity. CONCLUSIONS: In this study, influenza vaccination was associated with decreased mortality and influenza infections in community dwelling elderly. Our results indicate that, in a season of mild influenza activity and good antigenic match between vaccine strains and circulating strains, influenza vaccination reduced mortality in the vaccinated population. Our data support an annual vaccination strategy for all community-dwelling elderly. PMID- 12742809 TI - The tattooing paradox: are studies of acute hepatitis adequate to identify routes of transmission of subclinical hepatitis C infection? AB - BACKGROUND: The Hepatitis Branch of the Centers for Disease Control and Prevention does not recommend routine regulation and inspection of tattoo parlors because surveillance of hepatitis C virus (HCV)-positive acute hepatitis cases rarely identifies tattooing in the incubation period. However, the majority of seroepidemiological studies agree that tattooing is a strong, independent risk factor for subclinical HCV seropositivity. We postulated that this paradox might be explained if transmission of HCV by tattooing generally caused subclinical HCV seropositivity without the acute hepatitis syndrome. METHODS: We reanalyzed data from a prior seroepidemiological study of 626 consecutive patients who were unaware of their HCV serologic status and whose risk factors were ascertained by interview of an internist. Separate multiple logistic regression models were developed to predict a history of the acute hepatitis syndrome and HCV seropositivity. RESULTS: A history of injection-drug use was strongly associated with both HCV seropositivity (adjusted odds ratio [AOR], 7.2; 95% confidence interval [CI], 3.1-16.5) and a history of acute hepatitis (AOR, 5.9; 95% CI, 2.5 13.8), whereas having a commercially applied tattoo was strongly associated with HCV seropositivity (AOR, 6.5; 95% CI, 2.9-14.4) but not with a history of acute hepatitis (AOR, 1.2; 95% CI, 0.5-3.3). CONCLUSIONS: Intravenous injection of relatively large quantities of innocula of HCV may be more likely to result in the relatively rare acute HCV hepatitis syndrome, whereas intradermal exposure to small quantities of innocula may cause only subclinical HCV infections. If so, public policy on regulation and inspection of tattoo parlors should be determined by seroepidemiological studies rather than by the Sentinel Counties Study of acute hepatitis cases. PMID- 12742810 TI - Risk of cardiovascular disease-related and all-cause death according to serum concentrations of enterolactone: Kuopio Ischaemic Heart Disease Risk Factor Study. AB - BACKGROUND: Enterolactone is a plant-derived compound that has been associated with a reduced risk of acute coronary events and cancer. Several studies have suggested that serum enterolactone concentration may play a role as a biomarker of a diet high in fiber and vegetables. Owing to its phenolic structure, enterolactone and its plant lignan precursors, which are converted by intestinal bacteria to enterolactone, are potential antioxidants. METHODS: The associations between serum enterolactone level and the risk of coronary heart disease (CHD) related, cardiovascular disease (CVD)-related, and all-cause mortality were investigated in the Kuopio Ischaemic Heart Disease Risk Factor Study, which is a prospective population-based study of middle-aged Finnish men. The serum enterolactone concentration and cardiovascular risk factors were determined in 1889 men aged 42 to 60 years. In an average follow-up of 12.2 years, 70 CHD related, 103 CVD-related, and 242 all-cause deaths occurred in participants free of prior CVD. RESULTS: Multivariate analyses showed significant associations between elevated serum enterolactone concentration and reduced risk of CHD- and CVD-related mortality, but weaker associations in relation to all-cause mortality. In the Cox proportional hazards regression model adjusting for the most potent confounding factors, the risk of CHD-related (P =.03 for trend) and CVD-related (P =.04 for trend) death decreased linearly across quartiles of serum enterolactone concentration. CONCLUSIONS: Our data suggest that a high serum enterolactone level is associated with reduced CHD- and CVD-related mortality in middle-aged Finnish men. These results add to the evidence supporting the importance of whole grain foods, fruits, and vegetables in the prevention of premature death from CVD. PMID- 12742811 TI - Role of family history in identifying women with thrombophilia and higher risk of venous thromboembolism during oral contraception. AB - BACKGROUND: Unrecognized thrombophilic defects increase the risk of venous thromboembolism (VTE) in women during oral contraception (OC). We evaluated the sensitivity and specificity of a family history of VTE to identify thrombophilia in women before OC and after venous thrombotic complications during OC. METHODS: Thrombophilia screening was performed after obtaining a family history by means of a standardized questionnaire in (1) thrombosis-free women before OC and (2) women after an episode of VTE during OC. RESULTS: We evaluated 479 thrombosis free women before OC (age range, 15-49 years); family history was positive in 49 (10.2%). Thrombophilic defects were identified in 36 participants (7.5%; 95% confidence interval [CI], 5%-10%), 3 of whom had a positive family history (8.3%). The sensitivity and positive predictive value of family history of thrombophilic defects were 8.3% (95% CI, 2%-22%) and 6.1% (95% CI, 1%-17%), respectively. We also evaluated 189 women after VTE complications during OC (age range, 15-49 years); family history was positive in 48 (25.4%; 95% CI, 19%-32%), 22 of whom had a thrombophilic defect (45.8%; 95% CI, 31%-61%). Thrombophilic defects were identified in 81 women (42.8%; 95% CI, 36%-50%). The sensitivity and positive predictive value of family history of thrombophilic defects were 27.2% (95% CI, 18%-38%) and 45.8% (95% CI, 31%-61%), respectively. CONCLUSION: Family history of VTE has low sensitivity and positive predictive value for identifying women with thrombophilia who are more susceptible to VTE complications during OC. PMID- 12742812 TI - Right ventricular infarction in a patient with acute pulmonary embolism and normal coronary arteries. PMID- 12742813 TI - Malpractice claims regarding corticosteroids. PMID- 12742814 TI - Broadening the business case for patient safety. PMID- 12742815 TI - Acetaminophen and hypertension: a causal association or pain mediated? PMID- 12742816 TI - Hypertension and obesity. PMID- 12742817 TI - Analgesics and hypertension: causality or correlation? PMID- 12742818 TI - NSAIDs and hypertension. PMID- 12742819 TI - Chasing the wave. Unfashionable but important new concepts in arterial wave travel. PMID- 12742820 TI - Prologue: kinins and related systems. New life for old discoveries. PMID- 12742821 TI - Expression levels strongly affect ligand-induced sequestration of B2 bradykinin receptors in transfected cells. AB - Transfection of cells with expression vectors is one of the most important tools used to assess the effects of receptor mutations on ligand-induced receptor sequestration. Most transfection methods give rise to transiently or stably transfected clones with a wide range of receptor expression levels that may also depend on the mutations made. It is, therefore, important to determine how the regulation of the receptors depends on their numbers per cell. In Chinese hamster ovary (CHO) and human embryonic kidney (HEK)-293 cells expressing high levels of B(2) kinin receptors, we observed poor sequestration indicated by <20% reduction in cell surface receptor number after 10 min of stimulation with 1 microM bradykinin (BK) compared with >70% in low-expressing cells. Whereas the rate of [(3)H]BK internalization (internalized [(3)H]BK in percentage of total bound [(3)H]BK) in low-expressing cells was independent of the ligand-concentration used, in high-expressing cells a strong rate decrease was observed with higher (>1 nM) concentrations. Lower ligand concentrations, however, led to internalization rates identical to those obtained in low-expressing cells. Transiently transfected HEK and COS-7 cells showed results similar to those of stably high-expressing cells. Our results demonstrate the difficulty in determining the internalization pattern of (mutated) B(2) kinin receptors, and possibly of G protein-coupled receptors in general, using a sequestration assay in high-expressing cells or transiently transfected cells with high numbers of receptors per transfected cell. However, the receptor (mutant)-specific internalization rate can be measured, provided that the ligand concentrations used are below a threshold at which the internalization rate is still independent of the ligand concentration. PMID- 12742823 TI - Upregulation of Cdc2 and cyclin A during apoptosis of endothelial cells induced by cleaved high-molecular-weight kininogen. AB - We (8) reported that the cleaved high-molecular-weight kininogen (HKa) and its domain 5 (D5) inhibited angiogenesis. Further studies (15) revealed that D5 could inhibit cell proliferation and induce apoptosis of proliferating endothelial cells, which together may represent a critical part of antiangiogenic activity of HKa and D5. In the present study, we further examined the effect of HKa on cell cycle progression and cell viability. We report that HKa induced a significant upregulation of Cdc2 and cyclin A in proliferating endothelial cells, concurrent with a marked increase of Cdc2 activity. The increased expression of Cdc2 and cyclin A by HKa was not associated with an apparent change in cell cycle profiles of basic fibroblast growth factor-stimulated proliferating cells, but closely correlated with a marked increase of apoptosis, suggesting that the elevated Cdc2 activity is involved in HKa-induced apoptosis of proliferating endothelial cells. Our results support an emerging hypothesis that Cdc2 and cyclin A are important regulators for cell cycle as well as for apoptosis. PMID- 12742822 TI - Downregulation of bradykinin B2 receptor in human fibroblasts during prolonged agonist exposure. AB - Sustained activation of G protein-coupled receptors results in an attenuation of cellular responses, a phenomenon termed desensitization. Whereas mechanisms for rapid desensitization of ligand-receptor-G protein-effector systems are relatively well characterized, much less is known about long-term adaptation processes that occur in the continuous presence of an agonist. Here we have studied the fate of endogenously expressed bradykinin B(2) receptors on human fibroblasts during prolonged agonist treatment. Stimulation with bradykinin for up to 24 h resulted in a 50% reduction of surface binding sites that was paralleled by a similar decrease of total B(2) receptor protein followed by Western blotting using monoclonal antibodies to the B(2) receptor. Whereas B(2) receptor mRNA levels did not change during 24 h of agonist treatment, B(2) receptor de novo synthesis was attenuated by 35-50%, indicating translational control of B(2) receptor levels. Furthermore, the half-life of B(2) receptor protein was shortened by 20-40% as shown by (35)S-labeled pulse-chase and immunoprecipitation experiments. This study demonstrates that bradykinin B(2) receptor expression during long-term agonist treatment is primarily regulated on the (post)translational level, i.e., by attenuation of de novo synthesis and by reduction of receptor stability. PMID- 12742824 TI - Effects of varying impulse number on cotransmitter contributions to sympathetic vasoconstriction in rat tail artery. AB - We examined the contributions of the cotransmitters norepinephrine (NE), ATP, and neuropeptide Y (NPY) to sympathetically evoked vasoconstriction in the rat tail artery in isolated vascular rings by using 1-100 stimulation impulses at 20 Hz. Phentolamine (2 microM), the alpha-adrenoceptor antagonist, markedly reduced responses to all stimuli, although responses to lower impulse numbers were reduced less than responses to longer trains. The purinergic receptor antagonist suramin (100 microM) reduced all responses, but to a much greater extent with few impulse trains. Responses were further reduced or abolished by addition of the second antagonist. Any remaining responses were abolished by the NPY-Y(1) receptor antagonist BIBP-3226 (75 nM). NPY had a direct agonist action and potentiated sympathetically mediated responses. NPY (75 nM) potentiated responses and BIBP-3226 decreased responses to 2- and 20-impulse trains. Both affected responses from 2 impulses to >20 impulses, but there was no preferential effect on purinergic contributions to responses because neurally released NPY potentiated both "pure" NE and ATP responses equally. We conclude that all three cotransmitters contribute significantly to vascular responses and their contribution varies markedly with impulse numbers. There is considerable synergy between cotransmitters, especially with lower impulse numbers where NPY contributions are greater than expected. PMID- 12742825 TI - Rho-Rho kinase pathway is involved in the regulation of myogenic tone and pump activity in isolated lymph vessels. AB - To evaluate whether or not Rho-Rho kinase pathway is involved in the regulation of mechanical activity of lymph vessels, effects of Y-27632 and okadaic acid on lymph pump activity and myogenic, pressure- and agonist-induced tone were examined in isolated rat lymph vessels. Y-27632 caused a significant dilation with a cessation of the lymph pump activity. Y-27632 also produced a dose-related dilation of the lymph vessels precontracted by norepinephrine (NE)-, U-46619- or 80 mM KCl. Okadaic acid significantly constricted the lymph vessels and reduced the frequency of the lymph pump activity. Okadaic acid also produced a dose related constriction of the lymph vessels precontracted by NE or U-46619. The Y 27632-induced decrease of the frequency of lymph pump activity was significantly reversed by the pretreatment with okadaic acid. In the presence of Y-27632, the pressure-mediated tone of the lymph vessel was significantly decreased. On the other hand, okadaic acid significantly increased the pressure-mediated tone. These findings suggest that Rho kinase and myosin phosphatase activity in lymphatic smooth muscles may contribute to the regulation of lymph pump activity and may be also involved in the control of myogenic pressure- and agonist-induced tone. PMID- 12742826 TI - Fumonisin-induced blockade of ceramide synthase in sphingolipid biosynthetic pathway alters aortic input impedance spectrum of pigs. AB - The sphingolipid signaling pathway appears to play an important role in regulating vascular tone. We examined the effect of fumonisin B(1), a fungal toxin in corn that blocks ceramide synthase in the sphingolipid signaling pathway, on the ascending aortic impedance spectrum of pigs. Sixteen pigs were fed culture material containing fumonisin B(1) (20 mg/kg body wt) (n = 7) or a control diet (n = 9) daily for 3 days and then instrumented under alpha chloralose anesthesia for measurement of ascending aortic pressure and flow. Fumonisin ingestion increased serum sphinganine and sphingosine concentrations. Fumonisin ingestion also decreased cardiac output and characteristic impedance and increased the frequency of the first minimum impedance modulus, systemic vascular resistance, and the terminal, first, and second harmonic reflection coefficients, without changing mean arterial pressure. Thus blockade of ceramide synthase is accompanied by decreased vascular tone in systemic conduit arteries and increased vascular tone in systemic resistance vessels. The results indicate that the sphingolipid signaling pathway influences vascular tone in alpha chloralose-anesthetized pigs. PMID- 12742827 TI - Sphingosine 1-phosphate and control of vascular tone. AB - Sphingosine-1-phosphate (S1P) is a platelet-derived lipid mediator that activates the endothelial isoform of nitric oxide synthase (eNOS) in endothelial cells. However, the role of S1P in endothelium-dependent vasodilation and the signaling pathways elicited by S1P in intact vessels are largely unknown. We found that S1P induces dose-dependent transient relaxation of isolated pressurized mesenteric arterioles (EC(50) 10 +/- 3 nM); maximal vasodilation (55 +/- 8%) is seen approximately 2 min after S1P addition and returns to baseline by 5 min. S1P promotes comparable responses in arterioles from wild-type but not eNOS(null) mice. S1P-induced vasodilation is abrogated by removal of endothelium or by the addition of the NOS inhibitor N(omega)-monomethyl-l-arginine but is not affected by the cyclooxygenase inhibitor indomethacin, nor by the blockade of K(+) channels by using 4-aminopyridine. S1P-induced vasodilation is attenuated by pertussis toxin, by the phosphoinositide 3-kinase (PI3-kinase) inhibitor wortmannin, and by the calcium chelator BAPTA. With the use of high-sensitivity protein immunoblots in extracts from single pressurized vessels, we found that S1P, but not bradykinin, promotes the phosphorylation of eNOS at Ser(1179). Maximum S1P-induced eNOS Ser(1179) phosphorylation was reached at the time of maximum vasorelaxation, but enzyme phosphorylation persisted for several minutes after vasodilation had resolved. Thus regulatory pathways distinct from eNOS Ser(1179) dephosphorylation serve to terminate agonist-promoted vasorelaxation. Taken together, our findings demonstrate that S1P, an important intercellular mediator of platelet-vessel wall interactions, is a effective arteriolar vasodilator that acts via G protein-dependent, calcium-sensitive, and PI3-kinase modulated signaling pathways. PMID- 12742828 TI - Regulation of cardiac beta 1-adrenergic receptor transcription during the developmental transition. AB - The beta(1)-adrenergic receptor (beta(1)AR) gene contains binding sites for myc/max proteins within a glucocorticoid response element. Transcriptional activation of the beta(1)AR is the result of cooperative binding between c-myc and the glucocorticoid receptor on the beta(1)AR promoter. The transcriptional regulation of both beta(1)AR and c-myc are developmentally regulated. We used transcription rate assays of nuclei isolated from fetal hearts to demonstrate a fivefold increase in the transcription rate of beta(1)AR vs. postnatal hearts (P < 0.01). This was associated with a fourfold increase in c-myc transcription. Transcription rate assays performed in a rat fibroblast cell line that overexpresses c-myc (myc(+/+)) showed similarly increased beta(1)AR expression compared with the wild-type cell line. Transient transfection experiments in the myc(+/+) cells demonstrated robust expression of beta(1)AR promoter constructs, which was abrogated by mutation of the myc/max binding site or by cotransfection with a c-myc antisense expression vector. These results suggest that the regulation of cardiac beta(1)AR transcription and the expression of c-myc are tightly integrated. PMID- 12742829 TI - Appearance of an erythrocyte population with decreased deformability and hemoglobin content following sepsis. AB - With the use of the cecal ligation and puncture model in mice, this study tested whether sepsis-induced decreased erythrocyte deformability is restricted to a subpopulation of cells. Erythrocyte subpopulations were isolated by centrifugal elutriation. Lineweaver-Burk conversion of deformability-response curves to shear stress was used to determine the shear stress at half-maximal cell elongation (K(EI)) and maximal cell elongation (EI(max)). Sepsis decreased erythrocyte deformability in whole blood. K(EI) values were elevated (2.7 vs. 2.1 Pa) and EI(max) values decreased (0.56 vs. 0.50) in sepsis compared with sham mice. K(EI) values for cells eluted at 7 ml/min (smallest and oldest cells) were similar; however, K(EI) values for cells eluted at 8 ml/min were greater in septic than sham animals (2.50 vs. 2.10). Younger and larger subpopulations of erythrocytes (eluted at 9, 10, and 11 ml/min) also showed a tendency of decreased deformability in sepsis. Mean corpuscular hemoglobin content was decreased in cells eluted at 7 and 8 ml/min in sepsis (4.5 and 10.2 pg) compared to sham (7.4 and 11.4 pg) mice. This study indicates that an erythrocyte subpopulation that represents 20% of circulating cells shows the most pronounced decrease in cell deformability during sepsis. Increased rigidity together with decreased corpuscular hemoglobin content in these cells may contribute to microcirculatory dysfunction and immune modulation during sepsis. PMID- 12742830 TI - Muscle pump function during locomotion: mechanical coupling of stride frequency and muscle blood flow. AB - We imposed opposing oscillations in treadmill speed and grade on nine rats to test for direct mechanical coupling between stride frequency and hindlimb blood flow. Resting hindlimb blood flow was 15.5 +/- 1.7 ml/min. For 90 s at 7.5 m/min, rats alternated walking at -10 degrees for 10 s and +10 degrees for 10 s. This elicited oscillations in hindlimb blood flow having an amplitude of 4.1 +/- 0.5 ml/min (18% of mean flow) with a delay presumably due to metabolic vasodilation. Similar oscillations in speed (5.5-9.5 m/min) elicited oscillations in hindlimb blood flow (amplitude 3.4 +/- 0.5 ml/min, 15% of mean flow) with less of a delay, possibly due to changes in vasodilation and muscle pump function. We then simultaneously imposed these speed and grade oscillations out of phase (slow uphill, fast downhill). The rationale was that the oscillations in vasodilation evoked by the opposing oscillations in speed and grade would cancel each other, thereby testing the degree to which stride frequency affects hindlimb blood flow directly (i.e., muscle pumping). Opposing oscillations in speed and grade evoked oscillations in hindlimb blood flow having an amplitude of 3.3 +/- 0.6 ml/min (16% of mean flow) with no delay and directly in phase with the changes in speed and stride frequency. The finding that hindlimb blood flow changes directly with speed (when vasodilation caused by changes in speed and grade oppose each other) indicates that there is a direct coupling of stride frequency and hindlimb blood flow (i.e., muscle pumping). PMID- 12742831 TI - Increased expression of protein kinase C isoforms in heart failure due to myocardial infarction. AB - The activities of cardiac protein kinase C (PKC) were examined in hemodynamically assessed rats subsequent to myocardial infarction (MI). Both Ca(2+)-dependent and Ca(2+)-independent PKC activities increased significantly in left ventricular (LV) and right ventricular (RV) homogenates at 1, 2, 4, and 8 wk after MI was induced. PKC activities were also increased in both LV and RV cytosolic and particulate fractions from 8-wk infarcted rats. The relative protein contents of PKC-alpha, -beta, -epsilon, and -zeta isozymes were significantly increased in LV homogenate, cytosolic (except PKC-alpha), and particulate fractions from the failing rats. On the other hand, the protein contents of PKC-alpha, -beta, and epsilon isozymes, unlike the PKC-zeta isozyme, were increased in RV homogenate and cytosolic fractions, whereas the RV particulate fraction showed an increase in the PKC-alpha isozyme only. These changes in the LV and RV PKC activities and protein contents in the 8-wk infarcted animals were partially corrected by treatment with the angiotensin-converting enzyme inhibitor imidapril. No changes in protein kinase A activity and its protein content were seen in the 8-wk infarcted hearts. The results suggest that the increased PKC activity in cardiac dysfunction due to MI may be associated with an increase in the expression of PKC alpha, -beta, and -epsilon isozymes, and the improvement of heart function in the infarcted animals by imidapril may be due to partial prevention of changes in PKC activity and isozyme contents. PMID- 12742832 TI - Changes of RBC aggregation in oxygenation-deoxygenation: pH dependency and cell morphology. AB - The effects of the oxygenation-deoxygenation process on red blood cell (RBC) aggregation were examined in relation to morphological changes in RBCs and the contribution of CO(2). A low-shear rheoscope was used to measure the rate of rouleaux (one-dimensional aggregate) formation in diluted autologous plasma exposed to gas mixtures with different Po(2) and Pco(2). RBC indexes and RBC suspension pH were measured for the oxygenated or the deoxygenated condition, and the cell shape was observed with a scanning electron microscope. In the oxygenation-deoxygenation process, the rate of rouleaux formation increased with rising pH of the RBC suspension, which was lowered in the presence of CO(2). The rate increased with increasing mean corpuscular hemoglobin concentration (thus the cells shrank), which increased with rising pH and decreased in the presence of CO(2). With rising pH, cell diameter increased and cell thickness decreased (thus the cell flattened). In addition, slight echinocytosis was induced in the presence of CO(2), and the aggregation was reduced by the morphological change. In conclusion, RBC aggregation in the oxygenation-deoxygenation process is mainly influenced by the pH-dependent change in the surface area-to-volume ratio of the cells, and the aggregation is modified by CO(2)-induced acidification and the accompanying changes in mean corpuscular hemoglobin concentration and cell shape. PMID- 12742833 TI - Bax ablation protects against myocardial ischemia-reperfusion injury in transgenic mice. AB - The role of the proapototic Bax gene in ischemia-reperfusion (I/R) injury was studied in three groups of mice: homozygotic knockout mice lacking the Bax gene (Bax(-/-)), heterozygotic mice (Bax(+/-)), and wild-type mice (Bax(+/+)). Isolated hearts were subjected to ischemia (30 min, 37 degrees C) and then to 120 min of reperfusion. The left ventricular developed force of Bax-deficient vs. Bax(+/+) hearts at stabilization and at 120 min of reperfusion was 1,411 +/- 177 vs. 1,161 +/- 137 mg and 485 +/- 69 vs. 306 +/- 68 mg, respectively. Superior cardiac function of Bax(-/-) hearts after I/R was accompanied by a decrease in creatine kinase release, caspase 3 activity, irreversible ischemic injury, and the number of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling-positive cardiomyocytes. Electron microscopic evaluation revealed reduced damage to mitochondria and the nuclear chromatin structure in Bax deficient mice. In the Bax(+/-) hearts, the damage markers were moderate. The superior tolerance of Bax knockout hearts to I/R injury recommends this gene as a potential target for therapeutic intervention in patients with severe and intractable myocardial ischemia. PMID- 12742834 TI - Nonlinear effects in subthreshold virtual electrode polarization. AB - Introduction of the virtual electrode polarization (VEP) theory suggested solutions to several century-old puzzles of heart electrophysiology including explanation of the mechanisms of stimulation and defibrillation. Bidomain theory predicts that VEPs should exist at any stimulus strength. Although the presence of VEPs for strong suprathreshold pulses has been well documented, their existence at subthreshold strengths during diastole remains controversial. We studied cardiac membrane polarization produced by subthreshold stimuli in 1) rabbit ventricular muscle using high-resolution fluorescent imaging with the voltage-sensitive dye pyridinium 4-[2-[6-(dibutylamino)-2-naphthalenyl]-ethenyl] 1-(3-sulfopropyl)hydroxide (di-4-ANEPPS) and 2) an active bidomain model with Luo Rudy ion channel kinetics. Both in vitro and in numero models show that the common dog-bone-shaped VEP is present at any stimulus strength during both systole and diastole. Diastolic subthreshold VEPs exhibited nonlinear properties that were expressed in time-dependent asymmetric reversal of membrane polarization with respect to stimulus polarity. The bidomain model reveals that this asymmetry is due to nonlinear properties of the inward rectifier potassium current. Our results suggest that active ion channel kinetics modulate the transmembrane polarization pattern that is predicted by the linear bidomain model of cardiac syncytium. PMID- 12742835 TI - Ectopic pacing at physiological rate improves postanoxic recovery of the developing heart. AB - Recently, rapid and transient cardiac pacing was shown to induce preconditioning in animal models. Whether the electrical stimulation per se or the concomitant myocardial ischemia affords such a protection remains unknown. We tested the hypothesis that chronic pacing of a cardiac preparation maintained in a normoxic condition can induce protection. Hearts of 4-day-old chick embryos were electrically paced in ovo over a 12-h period using asynchronous and intermittent ventricular stimulation (5 min on-10 min off) at 110% of the intrinsic rate. Sham (n = 6) and paced hearts (n = 6) were then excised, mounted in vitro, and subjected successively to 30 min of normoxia (20% O(2)), 30 min of anoxia (0% O(2)), and 60 min of reoxygenation (20% O(2)). Electrocardiogram and atrial and ventricular contractions were simultaneously recorded throughout the experiment. Reoxygenation-induced chrono-, dromo-, and inotropic disturbances, incidence of arrhythmias, and changes in electromechanical delay (EMD) in atria and ventricle were systematically investigated in sham and paced hearts. Under normoxia, the isolated heart beat spontaneously and regularly, and all baseline functional parameters were similar in sham and paced groups (means +/- SD): heart rate (190 +/- 36 beats/min), P-R interval (104 +/- 25 ms), mechanical atrioventricular propagation (20 +/- 4 mm/s), ventricular shortening velocity (1.7 +/- 1 mm/s), atrial EMD (17 +/- 4 ms), and ventricular EMD (16 +/- 2 ms). Under anoxia, cardiac function progressively collapsed, and sinoatrial activity finally stopped after approximately 9 min in both groups. During reoxygenation, paced hearts showed 1) a lower incidence of arrhythmias than sham hearts, 2) an increased rate of recovery of ventricular contractility compared with sham hearts, and 3) a faster return of ventricular EMD to basal value than sham hearts. However, recovery of heart rate, atrioventricular conduction, and atrial EMD was not improved by pacing. Activity of all hearts was fully restored at the end of reoxygenation. These findings suggest that chronic electrical stimulation of the ventricle at a near-physiological rate selectively alters some cellular functions within the heart and constitutes a nonischemic means to increase myocardial tolerance to a subsequent hypoxia-reoxygenation. PMID- 12742836 TI - A randomized trial of patching regimens for treatment of moderate amblyopia in children. AB - OBJECTIVE: To compare 2 hours vs 6 hours of daily patching as treatments for moderate amblyopia in children younger than 7 years. METHODS: In a randomized multicenter (35 sites) clinical trial, 189 children younger than 7 years with amblyopia in the range of 20/40 to 20/80 were assigned to receive either 2 hours or 6 hours of daily patching combined with at least 1 hour per day of near visual activities during patching.Main Outcome Measure Visual acuity in the amblyopic eye after 4 months. RESULTS: Visual acuity in the amblyopic eye improved a similar amount in both groups. The improvement in the visual acuity of the amblyopic eye from baseline to 4 months averaged 2.40 lines in each group (P =.98). The 4-month visual acuity was at least 20/32 and/or improved from baseline by 3 or more lines in 62% of patients in each group (P>.99). CONCLUSION: When combined with prescribing 1 hour of near visual activities, 2 hours of daily patching produces an improvement in visual acuity that is of similar magnitude to the improvement produced by 6 hours of daily patching in treating moderate amblyopia in children aged 3 to 7 years. PMID- 12742837 TI - Montelukast, a leukotriene receptor antagonist, in vernal keratoconjunctivitis associated with asthma. AB - OBJECTIVE: To evaluate the effects on signs and symptoms of a coexisting vernal keratoconjunctivitis in patients treated with oral montelukast sodium for asthma. METHODS: Twelve patients with vernal keratoconjunctivitis and asthma were enrolled in this pilot study. Topical eyedrops or any systemic treatment was discontinued for at least 7 days before montelukast treatment. Patients were asked to grade their ocular discomfort daily. The following signs and symptoms were also recorded and graded through medical examination at baseline,after 15 days of treatment, and 15 days after treatment discontinuation: physician evaluated tarsal and bulbar papillae, hyperemia, secretion, and chemosis; and patient-evaluated itching, burning, tearing, photophobia, foreign body sensation, secretion, and redness. Peak expiratory flow rate at 8 AM was also recorded. Samples were collected at the same time points for enzyme-linked immunosorbent assay measurement of leukotriene B4 in tears and leukotriene E4 in urine. RESULTS: Eight of the 10 patients evaluated reported a reduction in symptoms at the end of treatment. Montelukast treatment significantly decreased physician rated hyperemia, secretion, and chemosis as well as patient-rated burning, tearing, photophobia, secretion, and redness. Effects persisted 15 days after discontinuation of treatment. Clinical changes were associated with a significant increase in leukotriene B4 in tears and a significant decrease in leukotriene E4 in urine after 15 days of treatment. CONCLUSION: The significant and persistent reduction of ocular signs and symptoms in asthmatic patients with vernal keratoconjunctivitis treated for 15 days with montelukast strongly suggests the need for double-masked placebo-controlled trials to confirm the potential of this new treatment in vernal keratoconjunctivitis. PMID- 12742838 TI - Short-term effect of dorzolamide hydrochloride on central corneal thickness in humans with cornea guttata. AB - OBJECTIVE: To investigate the short-term effect of dorzolamide hydrochloride, a topical carbonic anhydrase inhibitor, on central corneal thickness in patients with cornea guttata. DESIGN AND METHODS: In this randomized, placebo-controlled, double-masked, 3-drug crossover study, 20 patients with cornea guttata (mean endothelial cell count, 1321 cells/mm2) and 8 healthy control subjects (mean endothelial cell count, 2483 cells/mm2) were included. Study medications included 2% dorzolamide hydrochloride (Trusopt 2% eye drops; Merck & Co Inc, Whitehouse Station, NJ), 0.9% saline solution (saline placebo), and a solution identical to the carrier substance of dorzolamide in Trusopt (carrier placebo). The study drugs were applied 4 times per day for 1 day only. Central corneal thickness measurements were performed using partial coherence interferometry on every study day at baseline and after 24 hours of study medication treatment. MAIN OUTCOME MEASURES: Change in central corneal thickness. RESULTS: The mean thickening in central corneal thickness within 24 hours in eyes with cornea guttata treated with dorzolamide, saline placebo, and carrier placebo was 12.0 micro m (95% confidence interval [CI], 7.0-17.1 microm), 0.6 micro m (95% CI, -1.0 to 2.2 microm), and 1.3 micro m (95% CI, -0.1 to 2.6 microm), respectively. CONCLUSION: Application of dorzolamide for 1 day results in a slight but statistically significant thickening of central corneal thickness in patients with cornea guttata. PMID- 12742839 TI - Efficacy of ketotifen fumarate 0.025% ophthalmic solution compared with placebo in the conjunctival allergen challenge model. AB - BACKGROUND: Ketotifen fumarate blocks histamine1 (H1) receptors, stabilizes mast cells, and acts as an eosinophil inhibitor (decreases chemotaxis and activation of eosinophils). OBJECTIVE: To assess the efficacy of ketotifen 0.025% ophthalmic solution in the prevention of symptoms of allergic conjunctivitis, using the conjunctival allergen challenge model. METHODS: This was a single-center, double masked, randomized, placebo-controlled, contralateral-eye comparison, allergen challenge trial conducted in the United States. Subjects were randomized to receive ketotifen 0.025% in one eye and placebo in the other. At visits 1 and 2, allergen challenges were performed to determine the allergen concentration eliciting a qualifying reaction for each subject. At the 3 subsequent visits, subjects received 1 drop of ketotifen 0.025% ophthalmic solution in one eye and vehicle solution as placebo in the other eye 15 minutes (visit 3), 6 hours (visit 4), and 8 hours (visit 5) before allergen challenge. The primary efficacy measure was the subject's rating of itching at 3, 7, and 10 minutes after challenge. RESULTS: Of the 89 subjects randomly assigned to masked trial medication at visit 3, 72 completed the study. At visits 3, 4, and 5, mean itching scores were significantly better for ketotifen-treated eyes at all postchallenge time points, compared with placebo (P<.001). Also at visits 3, 4, and 5, ketotifen was statistically superior to placebo in reducing ocular hyperemia at all postchallenge time points (P<.05). CONCLUSIONS: Ketotifen was safe and statistically effective in reducing ocular itching and hyperemia associated with allergic conjunctivitis. Ketotifen's rapid onset of action (within 15 minutes) and extended duration of action (at least 8 hours) make it a valuable treatment for allergic conjunctivitis. PMID- 12742840 TI - Ultrasonographic biomicroscopy, Scheimpflug photography, and novel provocative tests in contralateral eyes of Chinese patients initially seen with acute angle closure. AB - OBJECTIVES: To compare ocular biometry of the contralateral eyes of individuals seen with acute angle closure (AAC) with eyes of population-based control subjects, and to assess novel provocative tests to study the mechanism of AAC. DESIGN: Prospective case-control study. PARTICIPANTS: Chinese persons seen as incident cases of AAC and Chinese population-based controls. METHODS: Slitlamp assessment, ultrasonographic biomicroscopy, Scheimpflug photography, and provocative testing were performed. MAIN OUTCOME MEASURES: Ocular biometric parameters including anterior chamber depth, limbal anterior chamber depth, axial length, lens thickness, and radius of corneal curvature were obtained. Ultrasonographic biomicroscopy parameters that include the angle-opening distance at 500 micro m and the angle-recess area were noted. Scheimpflug photography produced a single measure of angle width. RESULTS: Contralateral eyes of cases of AAC had shorter axial lengths, shallower anterior chamber depths, thicker lenses, and steeper radii of corneal curvature (P<.01). After adjusting for age and sex, cases had a mean adjusted axial length that was 1.2 mm shorter, an optical anterior chamber depth that was 0.63 mm shallower (24% shallower than controls), and lenses that were, on average, 0.35 mm thicker (P<.01). Furthermore, using multiple logistic regression to adjust for age and sex, patients with primary angle-closure glaucoma were 19 times as likely to have a shallower limbal anterior chamber depth (25%; 95% confidence interval, 8.3-45.2). Adjusting for age and sex, the mean angle-opening distance at 500 microm was 0.14 U less for cases, with a mean of 0.26 U in controls, making the angle-opening distance at 500 microm, on average, 54% less among cases. Scheimpflug photographs revealed an adjusted angle width of 21.6 degrees for controls and 15.1 degrees for cases (P<.05). Dynamic testing showed that the angle of control eyes tended to shallow less when going from light to dark and tended to open more when given 1 drop of pilocarpine hydrochloride. CONCLUSIONS: Contralateral eyes of individuals having an AAC attack tend to be shorter and have more crowded anterior segments than those of healthy controls. These static measures of ocular biometry indicate why some individuals are predisposed to AAC. Dynamic measures of the response to luminance changes and pilocarpine therapy indicate that differential reactions to these stimuli are also associated with an AAC attack. PMID- 12742841 TI - Classification of visual field abnormalities in the ocular hypertension treatment study. AB - OBJECTIVES: (1) To develop a classification system for visual field (VF) abnormalities, (2) to determine interreader and test-retest agreement, and (3) to determine the frequency of various VF defects in the Ocular Hypertension Treatment Study. METHODS: Follow-up VFs are performed every 6 months and are monitored for abnormality, indicated by a glaucoma hemifield test result or a corrected pattern SD outside the normal limits. As of January 1, 2002, 1636 patients had 2509 abnormal VFs. Three readers independently classified each hemifield using a classification system developed at the VF reading center. A subset (50%) of the abnormal VFs was reread to evaluate test-retest reader agreement. A mean deviation was calculated separately for the hemifields as an index to the severity of VF loss. MAIN OUTCOME MEASURES: A 97% interreader hemifield agreement. RESULTS: The average hemifield classification agreement (between any 2 of 3 readers) for 5018 hemifields was 97% and 88% for the 1266 abnormal VFs that were reread (agreement between the first and second classifications). Glaucomatous patterns of loss (partial arcuate, paracentral, and nasal step defects) composed the majority of VF defects. CONCLUSION: The Ocular Hypertension Treatment Study classification system has high reproducibility and provides a possible nomenclature for characterizing VF defects. PMID- 12742843 TI - Risk of age-related macular degeneration in eyes with macular drusen or hyperpigmentation: the Blue Mountains Eye Study cohort. AB - OBJECTIVE: To quantify the 5-year risk of age-related macular degeneration (AMD) in eyes with different macular drusen characteristics (ie, size, type, location, and total area) or hyperpigmentation in a population-based cohort. METHODS: The Blue Mountains Eye Study examined 3654 residents during 1992-1994; 2335 (75.1% of survivors) were reexamined during 1997-1999. Retinal photographs were graded using the Wisconsin Age-Related Maculopathy Grading System. Incident AMD lesions were defined by development of neovascular AMD or geographic atrophy in eyes without these lesions at baseline (eyes at risk). Age-adjusted relative risks (RRs) were determined. Generalized estimating equation models were used to estimate odds ratios, adjusting for the correlation between eyes and other AMD risk factors. Main Outcome Measure Incidence of AMD. RESULTS: Of the 4634 eyes at risk, 52 (1.1%) developed neovascular or atrophic AMD lesions over 5 years. In right eyes, presence vs absence of the following macular signs predicted AMD: drusen that were 125 micro m or larger (13.9 vs 0.6%; age-adjusted RR, 5.7; 95% confidence interval [CI], 3.6-9.0), indistinct soft or reticular drusen (23.2% vs 0.4%; RR, 9.9; 95% CI, 6.4-15.4), total drusen area of half the disc area or more (31.4% vs 0.6%; RR, 13.5; 95% CI, 8.0-22.8), and hyperpigmentation (14.4% vs 0.5%; RR, 8.0; 95% CI, 5.4-11.9). After adjusting for age, sex, and smoking status, eyes with these signs at baseline had a high likelihood of developing AMD. Eyes with Age-Related Eye Disease Study categories 3 and 4 were 5 times more likely to develop AMD compared with eyes in categories 1 and 2. CONCLUSION: This study quantifies the 5-year risk of AMD in eyes with macular drusen and hyperpigmentation. PMID- 12742842 TI - Retinal microvascular abnormalities in patients treated with external radiation for graves ophthalmopathy. AB - BACKGROUND: A prospective study was conducted to determine if external ionizing radiation could favorably influence the orbital manifestations of Graves ophthalmopathy. Diabetes and untreated systemic hypertension were exclusion criteria. Radiation was directed to the orbits of 42 affected patients using 0.2 rad (20 Gy) delivered in 10 doses of 0.02 rad (2 Gy). Patients were periodically examined during a 3-year interval. OBJECTIVE: To report retinal microvascular abnormalities observed in our study cohort. METHODS: Fundus findings documented with ophthalmoscopy, stereoscopic color photography, and stereoscopic fluorescein angiography prior to radiation were compared with similarly documented findings approximately 3 years following radiation. RESULTS: Prior to orbital radiation, retinal microvascular abnormalities were identified in 2 patients. The abnormalities were present bilaterally in one patient and unilaterally in the other. During the course of the study, microvascular abnormalities developed de novo in the unaffected retina of the latter patient while the retinopathy in the fellow eye progressed. Retinal microvascular abnormalities and their sequelae developed de novo in both eyes in 2 more patients. In addition to the radiation, other confounding factors known to be associated with microvascular retinopathy (uveitis, inadequately controlled systemic hypertension, and borderline blood glucose levels) were identified among the 3 patients whose eyes developed new retinal microvascular abnormalities. CONCLUSIONS: Whether the retinal microvascular abnormalities observed in these patients were caused or aggravated by external beam irradiation cannot be precisely ascertained. However, the observed progression and de novo development of retinal microvascular abnormalities within 3 years of orbital radiation raise concern that 0.2 rad (20 Gy) delivered to the orbit in 10 doses of 0.02 rad (2 Gy) may aggravate existing retinal microvascular abnormalities or cause radiation retinopathy in some patients with Graves disease. These findings and the failure of external beam radiation with 0.2 rad (2000 cGy) to favorably affect Graves ophthalmopathy, as demonstrated in a previous study, have led us to discourage further treatment of Graves ophthalmopathy with radiation. PMID- 12742844 TI - A randomized clinical trial of a single dose of intravitreal triamcinolone acetonide for neovascular age-related macular degeneration: one-year results. AB - OBJECTIVE: To determine if a single intravitreal injection of 4 mg of triamcinolone acetonide in patients with classic choroidal neovascularization associated with age-related macular degeneration can safely reduce the risk of severe visual loss. METHODS: A double-masked, placebo-controlled, randomized clinical trial was performed in patients 60 years or older who had choroidal neovascularization with any classic component, a duration of symptoms of less than 1 year, and a visual acuity of 20/200 or better. Best-corrected visual acuity, intraocular pressure, and cataract grading were performed before the injection and then at 3, 6, and 12 months. MAIN OUTCOME MEASURE: The development of severe loss of vision (30 letters) by survival analysis on an intention-to treat basis. RESULTS: One hundred fifty-one eyes were randomized into the study, and follow-up data were obtained for 73 (97%) of the 75 eyes in the treated group and for 70 (92%) of the 76 eyes in the control group. There was no difference between the 2 groups for the development of severe visual loss during the first year of the study (log-rank chi 2(1) = 0.03, P =.90). In both groups, the 12 month risk of severe visual loss was 35%, with a hazard ratio of 1.05 (95% confidence interval, 0.59-1.86). The change in size of the neovascular membranes, however, was significantly less in eyes receiving triamcinolone than in those receiving placebo 3 months after treatment (P =.01), although no difference was noted after 12 months. After 12 months, treated eyes had a significantly higher risk of an elevated intraocular pressure (31/75 [41%] vs 3/76 [4%]; P<.001), but not of cataract progression (P =.29). CONCLUSIONS: A single dose of intravitreal triamcinolone had no effect on the risk of loss of visual acuity during the first year of the study in eyes with age-related macular degeneration and classic choroidal neovascularization, despite a significant antiangiogenic effect found 3 months after treatment. This biological effect warrants further study. PMID- 12742846 TI - Lack of an association of apolipoprotein E gene polymorphisms with familial age related macular degeneration. AB - BACKGROUND: Previously, the epsilon 4 allele of apolipoprotein E (APOE) was reported to have a significant association with a decreased risk of age-related macular degeneration (AMD). In addition, the epsilon 2 allele of APOE was reported to be possibly associated with an increased risk of AMD. OBJECTIVE: To determine if APOE polymorphisms, previously reported to be associated with AMD, affect its expression in medium to large families, as well as in unrelated patients with AMD. METHODS: The APOE genotype was determined by HhaI restriction digests of polymerase chain reaction-amplified products in a collection of 259 affected and 207 unaffected individuals from 56 AMD families. Genotypes were determined similarly in a set of 104 unrelated AMD patients and in 113 unaffected control subjects. Diagnosis of AMD was based on clinical examination and evaluation of fundus photographs. Evidence of an association between alleles of APOE and AMD in families was tested by the following 4 statistical methods: chi2 analysis of simple allele counting, logistic regression analysis adjusting for age, construction of likelihood ratios of haplotype frequencies, and the pedigree disequilibrium test. RESULTS: None of the statistical methods used showed a significant association between the common alleles of APOE and AMD in our collection of families or in the set of unrelated AMD patients. CONCLUSIONS: No evidence was found to support an association between AMD in medium to large families and the epsilon 4 or epsilon 2 alleles of APOE. Neither was any evidence found for an association of APOE polymorphisms with the set of unrelated patients with AMD. However, a trend for a decreased risk of AMD associated with APOE epsilon 4 was observed in the set of unrelated patients with a family history of AMD. PMID- 12742845 TI - Association of emphysema, gout, and inflammatory markers with long-term incidence of age-related maculopathy. AB - OBJECTIVE: To examine the relationship of 2 diseases associated with systemic inflammatory response, emphysema and gout, and selected markers of systemic inflammation with the 10-year incidence of age-related maculopathy. DESIGN: Population-based cohort study. PARTICIPANTS: We included persons aged 43 to 86 years at baseline examination from 1988 to 1990 living in Beaver Dam, Wis, of whom 3684 subjects participated in a 5-year follow-up examination and 2764 participated in a 10-year follow-up. METHODS: Standardized protocols for physical examination, blood collection, administration of a questionnaire, and stereoscopic color fundus photography to determine the presence of age-related maculopathy. Standard univariate and multivariate analyses were performed. MAIN OUTCOME MEASURES: Incidence and progression of age-related maculopathy. RESULTS: While controlling for age, sex, and other factors (history of heavy drinking or smoking, systolic blood pressure, and vitamin use), a higher white blood cell count at baseline was associated with the 10-year incidence of drusen 125 microm or greater in diameter (risk ratio [RR] per 10(6)/ microL = 1.10; 95% confidence interval [CI], 1.03-1.17), retinal pigment epithelial depigmentation (RR = 2.08; 95% CI, 1.01-1.16), and progression of age-related maculopathy (RR = 1.09; 95% CI, 1.03-1.15). A lower serum albumin level was associated with the incidence of exudative macular degeneration (RR per grams per deciliter = 0.31; 95% CI, 0.13 0.76). A history of emphysema at baseline was associated with the incidence of retinal pigment epithelial depigmentation (RR = 2.84; 95% CI, 1.40-5.78), increased retinal pigment (RR = 2.20; 95% CI, 1.11-4.35), and exudative macular degeneration (RR = 5.12; 95% CI, 1.63-16.06); a history of gout was associated with the incidence of pure geographic atrophy (RR = 3.48; 95% CI, 1.27-9.53). CONCLUSIONS: These findings indicate modest relationships between both increased white blood cell count and emphysema and the increased 10-year incidence of lesions defining early and late age-related maculopathy. Further investigation of these relationships in other studies is needed. PMID- 12742847 TI - Soccer-related ocular injuries. AB - OBJECTIVE: To outline the severity and long-term sequelae of eye injuries in soccer. DESIGN: Prospective observational study of 163 patients who sustained soccer-related ocular injuries between April 1, 1992, and March 31, 2000 (8 years). METHODS: Patients were observed at a sports ophthalmology unit located in the largest university hospital of the northern region of the country and central to all major soccer fields in town. The data were recorded using the United States Eye Injury Registry report forms for initial and follow-up observation. MAIN OUTCOME MEASURES: (1) Self-reported history surrounding the ocular trauma, initial visual acuity, diagnosis, and operations and (2) final visual acuity, late diagnosis, and additional operations. RESULTS: Injuries occurred predominantly in young men (mean +/- SD age, 23.2 +/- 8.8 years) practicing indoor soccer (50.9%) or outdoor soccer (47.2%), and most resulted from a kicked ball (79.1%) near the goal post (60.1%). Angle recession and peripheral vitreoretinal lesions were more likely to occur in the superotemporal quadrant (54.7%; 95% confidence interval, 44.2%-65.0%; and 57.6%; 95% confidence interval, 48.4%-66.4%; respectively). Vitreoretinal lesions were present in 42.2% (95% confidence interval, 33.1%-51.8%) of patients with "normal" visual acuity (> or =20/40) and in 50.0% (95% confidence interval, 38.1%-61.8%) of patients without hyphema. No significant association was found between severity of injury and age, sex, type of soccer, level of athletic expertise, or player position. CONCLUSIONS: Severe ocular lesions can occur in soccer players without symptoms and at all skill levels. The development of laboratory models will be essential to explain the tendency for lesions to be in the superotemporal quadrant. The data support the need for protective eyewear designed specifically for soccer. PMID- 12742849 TI - A new fundus camera technique to help calculate eye-camera magnification: a rapid means to measure disc size. AB - OBJECTIVE: To find a simple means for calculating eye-camera magnification to permit estimation of true retinal object size from a retinal photograph. METHODS: The position of the focusing knob on 3 different retinal cameras (TRC-50F and TRC 50X; Topcon America Corp, Paramus, NJ; and the CR6-45NM Non-mydriatic Retinal Camera; Canon Inc, Tokyo, Japan) was measured during optic nerve photography and correlated with the refractive error, or spectacle refraction, of the subject (N = 11 for each camera). RESULTS: A strong correlation was found between focusing knob position and spectacle refraction for each of the 3 cameras tested (r = 0.96, r = 0.99, and r = 0.97, respectively). CONCLUSIONS: The focusing knob position reflects the spectacle refraction of the eye being photographed, and spectacle refraction is known to correlate well with eye-camera magnification. Therefore, focusing knob position can be used to help calculate eye-camera magnification and, hence, true retinal object size. CLINICAL RELEVANCE: The true size of the optic nerve head is important for the diagnosis of glaucoma from a retinal photograph. This technique is a simple means to calculate optic nerve head size, which may be especially useful in mass retinal photographic screening programs. PMID- 12742848 TI - Enhanced visualization of macular pathology with the use of ultrahigh-resolution optical coherence tomography. AB - OBJECTIVES: To demonstrate a new generation of ophthalmic optical coherence tomography (OCT) technology with unprecedented axial resolution for enhanced imaging of intraretinal microstructures and to investigate its clinical feasibility to visualize intraretinal morphology of macular pathology. METHODS: A clinically viable ultrahigh-resolution ophthalmic OCT system was developed and used in clinical imaging for the first time. Fifty-six eyes of 40 selected patients with different macular diseases including macular hole, macular edema, age-related macular degeneration, central serous chorioretinopathy, epiretinal membranes, and detachment of pigment epithelium and sensory retina were included. OUTCOME MEASURES: Ultrahigh-resolution tomograms visualizing intraretinal morphologic features in different retinal diseases. RESULTS: An axial image resolution of approximately 3 micro m was achieved in the eyes examined, nearly 2 orders of magnitude better than conventional ophthalmic ultrasound. Ultrahigh resolution OCT images provided additional diagnostically important information on intraretinal morphologic features that could not have been obtained by standard techniques. CONCLUSIONS: Ultrahigh-resolution ophthalmic OCT enables unprecedented visualization of intraretinal morphologic features and therefore has the potential to contribute to a better understanding of ocular pathogenesis, as well as to enhance the sensitivity and specificity for early ophthalmic diagnosis and to monitor the efficacy of therapy. This study establishes a baseline for the interpretation of ultrahigh-resolution ophthalmic OCT imaging of macular diseases. PMID- 12742850 TI - National preparedness for biological warfare and bioterrorism: smallpox and the ophthalmologist. PMID- 12742851 TI - Measuring outcomes in chronic diseases: ready for prime time? PMID- 12742852 TI - The ocular complications of smallpox and smallpox immunization. AB - Although smallpox was eradicated worldwide, concerns have been raised about the use of smallpox as a biological weapon. Plans are being considered for smallpox immunization in the United States. Variola virus, the cause of smallpox, and vaccinia virus, used in smallpox immunization, are both orthopoxviruses that are associated with serious ocular complications, including eyelid and conjunctival infection, corneal ulceration, disciform keratitis, iritis, optic neuritis, and blindness. About 5% to 9% of patients with smallpox develop ocular complications, and case-fatality rates reach 20% to 35% among unvaccinated individuals. About 10 to 20 patients develop ocular complications per 1 million smallpox immunizations, usually through autoinoculation, in which the patient transfers vaccinia from the immunization site to the eye. The risk of ocular vaccinia infection may be reduced by instructing patients and individuals in close contact with the vaccinee to wash their hands often and avoid touching the immunization site and their eyes. Topical antiviral therapy, topical steroids, and topical and oral antibiotics have been used to reduce the ocular complications of smallpox immunization. In contrast, there has been little experience with the use of these therapies for the ocular complications of smallpox. PMID- 12742853 TI - An Indian adventure: Derrick Vail's Shikarpur fragments. PMID- 12742854 TI - Ultrasound biomicroscopy in management of malignant iris melanoma. PMID- 12742855 TI - Klippel-Trenaunay syndrome and rhabdomyosarcoma in a 3-year-old. PMID- 12742856 TI - Secondary chronic open-angle glaucoma after intravitreal triamcinolone acetonide. PMID- 12742857 TI - Bone located centrally within a dermolipoma. PMID- 12742858 TI - Macular hole secondary to fungal endophthalmitis. PMID- 12742859 TI - Rosai-Dorfman disease with bilateral serous retinal detachment. PMID- 12742860 TI - Voriconazole treatment of fungal scleritis and epibulbar abscess resulting from scleral buckle infection. PMID- 12742861 TI - Endothelial proliferation in diabetic retinal microaneurysms. PMID- 12742862 TI - Central serous chorioretinopathy imaged by optical coherence tomography. PMID- 12742863 TI - Episodic segmental iris dilator muscle spasm: the tadpole-shaped pupil. PMID- 12742864 TI - Pinel orders the chains removed from the insane at Bicetre. PMID- 12742865 TI - White matter changes in schizophrenia: evidence for myelin-related dysfunction. AB - Numerous lines of inquiry implicate connectivity as a central abnormality in schizophrenia. Myelination and factors that affect myelination, such as the function of oligodendroglia, are critical processes that could profoundly affect neuronal connectivity, especially given the diffuse distribution of oligodendrocytes and the widespread distribution of brain regions that have been implicated in schizophrenia. Multiple lines of evidence now converge to implicate oligodendroglia and myelin in schizophrenia. Imaging and neurocytochemical evidence, similarities with demyelinating diseases, age-related changes in white matter, myelin-related gene abnormalities, and morphologic abnormalities in the oligodendroglia demonstrated in schizophrenic brains are all examined in light of the hypothesis that oligodendroglial dysfunction and even death, with subsequent abnormalities in myelin maintenance and repair, contribute to the schizophrenic syndrome. PMID- 12742866 TI - Placebo or active control trials of antipsychotic drugs? AB - The placebo-controlled trial has been the standard method to demonstrate efficacy and safety of antipsychotic drugs. We reviewed the scientific and ethical advantages and disadvantages of the placebo-controlled trial and an alternative method, the active-control trial, focusing more specifically on the active control noninferiority trial. Recent meta-analyses indicate that a therapeutic dose of second-generation antipsychotic will very likely be statistically superior to placebo in an adequate trial, and that the average improvement of schizophrenia symptoms in a placebo arm will be small. These findings strengthen the scientific and ethical justification for the active-control noninferiority trial. New drugs in the pharmacotherapy for schizophrenia are often claimed to differ from their marketed competitors in their safety profile rather than in antipsychotic efficacy. Thus, in many cases, it appears sufficient to demonstrate mere noninferiority (rather than superiority) of antipsychotic efficacy in comparison with a standard antipsychotic. The active-control noninferiority trial is suitable for such demonstration. Sample size requirements for various equivalence margins in noninferiority trials are provided. Scientific and ethical arguments should lead to a more frequent use of the active-control noninferiority trial design. PMID- 12742867 TI - Association of serum antibodies to herpes simplex virus 1 with cognitive deficits in individuals with schizophrenia. AB - BACKGROUND: Cognitive deficits are a characteristic feature of schizophrenia and contribute to the profound disabilities associated with this illness. Some of the cognitive deficits that occur in individuals with schizophrenia are similar to those found in individuals who have recovered from central nervous system infections with human herpesviruses. METHODS: We measured cognitive functioning and serologic evidence of infection with human herpesviruses in 229 outpatients with schizophrenia. We evaluated cognitive functioning with the Repeatable Battery for the Assessment of Neuropsychological Status. For each patient, serum IgG class antibodies with specificities for the following potentially neurotropic human herpesviruses were measured by means of a solid-phase immunoassay: herpes simplex viruses 1 and 2, cytomegalovirus, Epstein-Barr virus, human herpesvirus 6, and varicella-zoster virus. We determined the association between serologic evidence of herpesviruses infection and cognitive functioning by univariate and multivariate analyses, including demographic and clinical factors associated with cognitive functioning. RESULTS: We found that serologic evidence of infection with herpes simplex virus 1 is an independent predictor of cognitive dysfunction in individuals with schizophrenia. Discriminant function analysis indicated that much of the difference in cognitive functioning could be attributed to immediate memory. We found no significant association between cognitive dysfunction and serologic evidence of infection with other human herpesviruses. CONCLUSION: Serologic evidence of herpes simplex virus 1 infection is associated with cognitive impairment in schizophrenia. PMID- 12742868 TI - Prospective study of adult mental disturbance in offspring of women with psychosis. AB - BACKGROUND: The high-risk method is an important strategy for studying the antecedents and causes of schizophrenia and other psychoses. The Swedish High Risk Project is a prospective longitudinal study of offspring of women with a history of schizophrenic, schizoaffective, affective, or unspecified functional psychoses and control women with no history of psychosis. The offspring and their environments were studied beginning before birth, and again during childhood. This article reports the mental outcome results from the first adult follow-up at age 22 years. METHODS: Of 178 offspring, 166 (93%) were followed up and blindly assessed using standardized methods, including a self-report scale for mental symptoms and the Structured Clinical Interview for DSM-III-R. RESULTS: Compared with controls (n = 91), the offspring of mothers with schizophrenia (n = 28) showed a significantly increased frequency of DSM-III-R Axis I and Axis II disorders, poor global functioning, high Symptom Checklist-90 scores, and a history of mental health care and psychopharmacologic medication use. Offspring of mothers with affective disorders (n = 22) showed high Symptom Checklist-90 scores, more frequent poor functioning, and receipt of mental health care, with a significant increase in Axis I depressive disorders and no increase in Axis II disorders. The extension of schizophrenia and affective risk groups to include additional maternal "spectrum cases" (10 and 15 individuals, respectively) generally yielded similar results. CONCLUSIONS: Maternal schizophrenia is associated with widespread increases in offspring mental disturbance in adolescence and young adulthood, differing from offspring disturbance associated with maternal affective disorder. PMID- 12742869 TI - Randomized trial of trauma-focused group therapy for posttraumatic stress disorder: results from a department of veterans affairs cooperative study. AB - BACKGROUND: Department of Veterans Affairs Cooperative Study 420 is a randomized clinical trial of 2 methods of group psychotherapy for treating posttraumatic stress disorder (PTSD) in male Vietnam veterans. METHODS: Vietnam veterans (360 men) were randomly assigned to receive trauma-focused group psychotherapy or a present-centered comparison treatment that avoided trauma focus. Treatment was provided weekly to groups of 6 members for 30 weeks, followed by 5 monthly booster sessions. Severity of PTSD was the primary outcome. Additional measures were other psychiatric symptoms, functional status, quality of life, physical health, and service utilization. Follow-up assessments were conducted at the end of treatment (7 months) and at the end of the booster sessions (12 months); 325 individuals participated in 1 or both assessments. Additional follow-up for PTSD severity was performed in a subset of participants at 18 and 24 months. RESULTS: Although posttreatment assessments of PTSD severity and other measures were significantly improved from baseline, intention-to-treat analyses found no overall differences between therapy groups on any outcome. Analyses of data from participants who received an adequate dose of treatment suggested that trauma focused group therapy reduced avoidance and numbing and, possibly, PTSD symptoms. Dropout from treatment was higher in trauma-focused group treatment. Average improvement was modest in both treatments, although approximately 40% of participants showed clinically significant change. CONCLUSIONS: This study did not find a treatment effect for trauma-focused group therapy. The difference between the effectiveness and adequate dose findings suggests the possible value of methods to enhance the delivery of cognitive-behavioral treatments in clinical practice settings. PMID- 12742870 TI - A sib-pair study of the Temperament and Character Inventory scales in major depression. AB - BACKGROUND: Certain aspects of the personality may be associated with the vulnerability to develop depression. A sib-pair method has been used to examine the familiality of the 7 scales of the Temperament and Character Inventory (TCI) and whether this could be related to the genetic vulnerability to develop depression. METHODS: Probands with depression and their nearest-aged siblings from Wales were compared with healthy control probands and their nearest-aged siblings on the TCI and measures of depressed mood. RESULTS: All 7 scales of the TCI were familial, and scores on 6 of the scales were similar to US population scores. However, the Welsh subjects' scores on the self-transcendence scale were markedly lower than the US mean, suggesting strong cultural or national influences on this measure. Harm avoidance scores were substantially influenced by current and past depression, but this scale also showed stable traitlike characteristics that are likely related to the genetic vulnerability to depression. Novelty seeking and self-directedness were also partly state dependent and were negatively correlated with low mood; high scorers may be resilient to the development of depression. High reward dependence may also protect against the development of depression and is unrelated to mood state. The cooperativeness, persistence, and self-transcendence scales appear to have a limited relationship with the development of depression. CONCLUSIONS: Harm avoidance, reward dependence, novelty seeking, and self-directedness have traitlike characteristics that are related to the familiality of depression. Cooperativeness, self-transcendence, and persistence are also familial, but this appears to be unrelated to depression. PMID- 12742871 TI - The heritability of bipolar affective disorder and the genetic relationship to unipolar depression. AB - BACKGROUND: Twin studies of bipolar affective disorder (BPD) have either been small or have not used explicit diagnostic criteria. There has been little use of genetic model fitting and no analyses to explore the etiological overlap with unipolar depression (UPD). METHODS: Sixty-seven twin pairs, 30 monozygotic and 37 dizygotic, in which the proband had BPD were ascertained, and lifetime diagnoses were made using DSM-IV criteria. Univariate models were applied to estimate the contribution of additive genetic and environmental effects. Bipolar data were then combined with those from 68 monozygotic and 109 dizygotic pairs in which the proband had UPD. Two models were explored: a classic 2-threshold approach, in which BPD and UPD occupy the same continuum of liability but differ in severity, and a correlated liability model of mania and depression. RESULTS: Heritability of BPD was estimated at 85% (95% confidence interval [CI], 0.73-0.93) using narrow concordance and 89% (95% CI, 0.61-1.0) using broad concordance, with no shared environmental effects detected. A 2-threshold model was an unsatisfactory fit. Fitting a correlated liability model revealed a genetic correlation of 0.65 (95% CI, 0.58-0.75) between mania and depression and a correlation of 0.59 (95% CI, 0.15-0.84) for nonfamilial environment. Approximately 71% of the genetic variance for mania was not shared with depression. CONCLUSIONS: As defined by the DSM-IV, BPD is highly heritable. There are substantial genetic and nonshared environmental correlations between mania and depression, but most of the genetic variance in liability to mania is specific to the manic syndrome. PMID- 12742872 TI - Parent-child conflict and the comorbidity among childhood externalizing disorders. AB - BACKGROUND: Previous research has suggested that substantial comorbidity exists among childhood externalizing disorders, specifically attention deficit/hyperactivity disorder (ADHD), oppositional defiant disorder (ODD), and conduct disorder (CD). Moreover, parent-child conflict predicts each of these disorders. Our goals were to determine whether parent-child conflict was associated with the comorbidity among ADHD, CD, and ODD, and to explicitly examine the etiology of this association via a genetically informative design. METHODS: We compared the fit of the following 2 biometric models: the 2-factor common-pathway model, which examined genetic and environmental contributions to the relationship between conflict and the covariation among the 3 disorders, and the Cholesky model, which examined the relationship between conflict and each disorder individually. The sample consisted of 808 same-sex 11-year-old twin pairs from the Minnesota Twin Family Study, a population-based sample of Minnesota twins and their families. Main outcome measures included symptom counts for ADHD, CD, and ODD, obtained from structured interviews administered to twins and their mothers. Parent-child conflict was assessed via mother and twin reports of the Parental Environment Questionnaire. RESULTS: The 2-factor model provided a better fit to the data. These results indicated that conflict accounted for 33% of the covariation among the disorders, via genetic and environmental factors. CONCLUSIONS: Parent-child conflict appears to act as a common vulnerability that increases risk for multiple childhood disorders. Furthermore, this association is mediated via common genetic and environmental factors. These findings support the idea that the comorbidity among these disorders partially reflects core psychopathological processes in the family environment that link putatively separate psychiatric disorders. PMID- 12742873 TI - Treatment of trichotillomania with behavioral therapy or fluoxetine: a randomized, waiting-list controlled study. AB - BACKGROUND: Both behavioral therapy (BT) and serotonin reuptake inhibitors have been reported effective in the treatment of trichotillomania. This study examines the efficacy of BT and fluoxetine hydrochloride compared with a waiting-list (WL) control group. METHODS: Forty-three patients with trichotillomania entered a 12 week randomized, WL-controlled study of BT and fluoxetine (60 mg/d). Forty patients (14 in the BT group, 11 in the fluoxetine group, and 15 in the WL group) completed the trial. Treatment effects were evaluated using the Massachusetts General Hospital Hairpulling Scale, and severity of hair loss was rated by independent assessors. In addition, we measured general symptoms of psychopathologic abnormalities and depression. RESULTS: For reducing the symptoms of trichotillomania, BT was superior. Patients in the BT group showed a significantly greater reduction in trichotillomania symptoms, higher effect sizes (Massachusetts General Hospital Hairpulling Scale: BT, 3.80; fluoxetine, 0.42; and WL, 1.09), and more clinically significant changes (BT, 64%; fluoxetine, 9%; and WL, 20%) than patients in the fluoxetine and WL groups. For severity of hair loss, a similar trend was also found in favor of the BT group. No significant differences between groups were established for general psychopathologic and depressive symptoms. CONCLUSIONS: Behavioral therapy is highly effective for reducing symptoms of trichotillomania in the short term, whereas fluoxetine is not. PMID- 12742874 TI - Autistic traits in the general population: a twin study. AB - BACKGROUND: Recent research has indicated that autism is not a discrete disorder and that family members of autistic probands have an increased likelihood of exhibiting autistic symptoms with a wide range of severity, often below the threshold for a diagnosis of an autism spectrum disorder. OBJECTIVE: To examine the distribution and genetic structure of autistic traits in the general population using a newly established quantitative measure of autistic traits, the Social Responsiveness Scale (formerly known as the Social Reciprocity Scale). METHODS: The sample consisted of 788 pairs of twins aged 7 to 15 years, randomly selected from the pool of participants in a large epidemiologic study (the Missouri Twin Study). One parent of each pair of twins completed the Social Responsiveness Scale on each child. The data were subjected to structural equation modeling. RESULTS: Autistic traits as measured by the Social Responsiveness Scale were continuously distributed and moderately to highly heritable. Levels of severity of autistic traits at or above the previously published mean for patients with pervasive developmental disorder not otherwise specified were found in 1.4% of boys and 0.3% of girls. Structural equation modeling revealed no evidence for the existence of sex-specific genetic influences, and suggested specific mechanisms by which females may be relatively protected from vulnerability to autistic traits. CONCLUSIONS: These data indicate that the social deficits characteristic of autism spectrum disorders are common. Given the continuous distribution of these traits, it may be arbitrary where cutoffs are made between research designations of being "affected" vs "unaffected" with a pervasive developmental disorder. The genes influencing autistic traits appear to be the same for boys and girls. Lower prevalence (and severity) of autistic traits in girls may be the result of increased sensitivity to early environmental influences that operate to promote social competency. PMID- 12742875 TI - Learning to listen. PMID- 12742876 TI - Longitudinal care improves disclosure of psychosocial information. AB - BACKGROUND: While longitudinal primary care is thought to promote patient rapport and trust, it is not known if longitudinality helps overcome barriers to communication that may occur when the patient and physician are of different ethnicities and/or sexes. OBJECTIVE: To examine if longitudinal pediatric care ameliorates disparities in parent disclosure of psychosocial information associated with ethnic and gender discordance between parent and physician. DESIGN: Longitudinal, observational study of parent-physician interaction at early visits and over the course of 1 year. PARTICIPANTS: Parents (90% African American and 10% white mothers or female guardians) and their infant's assigned primary care physician (white first- and second-year pediatric residents). MAIN OUTCOME MEASURE: Parents' psychosocial information giving measured by the Roter Interaction Analysis System. RESULTS: Sex- and race-related barriers to disclosure of psychosocial information were evident early in the parent-physician relationship. At early visits, African American mothers made 26% fewer psychosocial statements than white mothers; this discrepancy was not affected by physician sex. At early visits, white mothers made twice as many psychosocial statements when seeing white female compared with white male physicians. CONCLUSIONS: Patient-centeredness is an important factor promoting psychosocial information giving for African American and white mothers, regardless of physician sex. Longitudinal relationships facilitate mothers' disclosure to physicians of a different ethnicity or sex, but only if physicians remain patient centered. PMID- 12742877 TI - Guidelines for Adolescent Preventive Services: the GAPS in practice. AB - DESIGN: Pre- and post-Guidelines for Adolescent Preventive Services (GAPS) comparison of outcomes gathered via chart audit. SETTING: A rural hospital-based general pediatric clinic. PARTICIPANTS: Adolescents who underwent annual examinations between April 1, 1998, and March 31, 2001. A random sample of 441 medical records was reviewed. INTERVENTION: Training in the GAPS model and use of the questionnaire began in April 1998. MAIN OUTCOME MEASURES: Detection of, discussion of, and referrals for GAPS-related risk behavior. RESULTS: The medical records of 162 younger adolescents (aged 11-15 years) and 279 older adolescents (aged 16-19 years) were audited. Detection of risk behaviors increased from 19% at baseline to 95% with the initial GAPS and 87% with the periodic GAPS. The most prevalent risk factor was having a rifle or gun in the home (younger adolescents, 47% and older adolescents, 39%). The mean number of risk behaviors and health concerns documented was higher in the initial GAPS (4.8 and 1.3, respectively) than in the periodic GAPS (3.8 and 0.7) (P =.01 and.006). The GAPS questionnaires detected lower levels of risk behavior compared with a local Youth Risk Behavior Survey. Controlling for sex, age, and clinician, discussion of psychosocial topics increased during the study period; however, there was considerable variation among clinicians regarding the topics addressed. The GAPS-related referral rate did not change significantly. CONCLUSION: The GAPS model increases clinicians' detection and discussion of risk behaviors. PMID- 12742878 TI - Effect of patient priming and primary care provider prompting on adolescent provider communication about alcohol. AB - OBJECTIVE: To determine whether priming adolescent patients to discuss alcohol with their primary care providers and prompting providers to discuss alcohol increases adolescent-provider communication about alcohol. DESIGN: Randomized controlled trial. SETTING: Five managed care group practices in Washington, DC. PARTICIPANTS: Consecutive patients aged 12 to 17 years who were seeing primary care providers (n = 26) for health checkups. Of 892 eligible adolescents, 444 (50%) were randomized and completed data collection. Most adolescents (80%) were African American, 55% were male, and 17% currently drank alcohol. INTERVENTION: Usual care (group 1) vs adolescent priming with alcohol self-assessment and education just before their health checkups (group 2) vs adolescent priming and provider prompting with the adolescent's self-assessment and a patient education brochure (group 3). MAIN OUTCOME MEASURES: This exploratory substudy of a longitudinal study on adolescent alcohol behaviors examined adolescent-provider communication by adolescent exit survey, researcher observation, and audiotapes of a subsample of visits. RESULTS: More adolescents in group 3 (96%) than group 1 (87%) reported that their provider talked about alcohol (adjusted odds ratio [OR], 1.10; 95% confidence interval [CI], 1.04-1.17). More adolescents in group 3 (18%) than group 1 (10%) reported asking about alcohol (adjusted OR, 1.08; 95% CI, 1.00-1.16). The mean +/- SD number of minutes adolescents were with their providers without parents being present was greater for group 3 (10.8 +/- 7.6) than group 1 (8.8 +/- 8.0). Adolescents in group 2 spent more time with their provider and reported initiating more discussion not specific to alcohol than did group 1 adolescents. CONCLUSION: Adolescent priming and provider prompting increases adolescent-provider communication about alcohol. PMID- 12742879 TI - Antibodies reactive to Rickettsia rickettsii among children living in the southeast and south central regions of the United States. AB - BACKGROUND: The reported annual incidence of Rocky Mountain spotted fever in the United States is 2.2 per million, but studies have suggested that human infection with Rickettsia rickettsii may be more common. This study estimated the prevalence of antibodies reactive to R rickettsii among children living in the southeastern and south central United States. STUDY DESIGN: Approximately 300 specimens were obtained from children at each of 7 pediatric referral centers (N = 1999). Serum was tested for R rickettsii antibodies by means of indirect immunofluorescence antibody assay. Three different cutoff titers (>or=64, >or=128, and >or=256) represented increasing levels of stringency to define positive specimens. RESULTS: Overall, 12.0% of children had R rickettsii antibody titers of at least 64; 7.3%, at least 128; and 4.3%, at least 256. Strong relationships were seen between increasing age and seroprevalence at each cutoff titer. Remarkably, 6.4% of children aged 13 to 17 years had titers of at least 256. Age-adjusted seroprevalence rates at titers of at least 64 varied from 21.9% in Little Rock, Ark, to 3.5% in Louisville, Ky. At titers of at least 256, seroprevalence ranged from 7.7% in Nashville, Tenn, to 1.8% in Winston-Salem, NC. Only site and age group were strong predictors of seropositivity; a weak association was seen with nonurban residence. CONCLUSIONS: To our knowledge, this is the largest serosurvey of rickettsial infection in children in the United States. Within the limitations of the immunofluorescence antibody assay, these data suggest that infections with R rickettsii or antigenically related spotted fever group rickettsiae may be common and subclinical. The results also have implications for the interpretation of single immunofluorescence antibody assay titers in children with suspected Rocky Mountain spotted fever. PMID- 12742880 TI - Does your child have asthma? Parent reports and medication use for pediatric asthma. AB - OBJECTIVE: To assess parental reporting of diagnosis used in surveys as an indicator of pediatric asthma prevalence. METHODS: Analysis of the Medical Expenditure Panel Survey, 1996 and 1997 (10 404 children aged from 0 to 17 years). All values are expressed as mean (SE). RESULTS: Asthma medications were purchased for 2.5% (0.2%) of children. Parents of 45.4% (4.0%) of these children failed to report asthma, including 41.3 (10.5%) of those for whom maintenance medications were purchased. These findings remained unchanged when very young children were excluded from the sample. Controlling for insurance coverage, no racial, ethnic, or socioeconomic disparities in reported asthma were found; however, poor children were more likely to have maintenance medications purchased (odds ratio, 4.9; 95% confidence interval, 2.3-10.4). CONCLUSIONS: Surveys of parental reports of asthma overlook many children with active disease. Dependence on parental reports may underestimate the prevalence of serious asthma among poor children. The parents in this study who fail to report asthma may represent a group that perceives their children's disease as less serious a problem despite active purchasing of medications. PMID- 12742881 TI - The effectiveness of assessment and referral on immunization coverage in the special supplemental nutrition program for women, infants, and children. AB - BACKGROUND: The use of immunization assessment and referral (A/R) in the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) has been shown to produce dramatic improvements in vaccination coverage when coupled with parental incentive; however, data are lacking to support the use of A/R alone. OBJECTIVE: To determine the effectiveness of A/R in increasing immunization coverage among WIC participants. DESIGN: Participating WIC centers were assigned to1 of 3 interventions that delivered A/R of varying frequency or a control group. SETTING: Twenty of the largest Public Health Foundation Enterprises-WIC centers in Los Angeles County. PARTICIPANTS: Children continuously enrolled in participating WIC centers from 6 to 24 months of age. INTERVENTION: Assessment of child's vaccination status followed by referral to a health care provider for those lacking indicated vaccinations. MAIN OUTCOME MEASURE: Up-to-date (UTD) status at 24 months of age for all recommended vaccines. RESULTS: Baseline coverage rates were similar among all study sites (overall, 77% UTD). After the study period, compared with the controls (88% UTD), we found no differences in immunization coverage among WIC centers that administered A/R at every visit (every 2 months) to all children (90% UTD; adjusted odds ratio [OR], 1.02; 95% confidence interval [CI], 0.54-1.94), every 6 months to all children (89% UTD; OR, 0.98; 95% CI, 0.62-1.56), or every visit to children found to be behind at 8 months of age (89% UTD; OR, 0.89; 95% CI, 0.48-1.68). CONCLUSION: In this urban population of WIC children with high baseline immunization coverage, A/R was not effective in increasing immunization coverage. PMID- 12742882 TI - Sleep disruption and objective sleepiness in children with beta-thalassemia and congenital dyserythropoietic anemia. AB - BACKGROUND: Sleep fragmentation and periodic leg movement syndrome (PLMS) have been reported in adults with iron deficiency anemia. Little is known about sleep function and daytime sleepiness in children with chronic anemia such as beta thalassemia or congenital dyserythropoietic anemia type 1 (CDA-1). OBJECTIVES: To investigate if children and adolescents who have beta-thalassemia (major or intermedia) or CDA-1 experience sleep fragmentation and objective daytime sleepiness and also to investigate if children and adolescents with beta thalassemia have obstructive sleep apnea. METHODS: Ten patients (7 males and 3 females) with beta-thalassemia (mean [SD] age, 10.4 [7.3] years), 10 patients (7 males and 3 females) with CDA-1 (mean [SD] age, 13.5 [5.1] years), and 13 healthy volunteer control children(7 males and 6 females) (mean [SD] age, 10 [4] years) underwent nocturnal polysomnographic studies. A multiple sleep latency test was performed for 6 patients who had beta-thalassemia and 8 patients who had CDA-1. RESULTS: Both patient groups, that is, those who had beta-thalassemia and those who had CDA-1, had multiple arousals during sleep (mean [SD], 27.8 [11.4] events per hour and 23.8 [11.8] events per hour, respectively) compared with the control subjects (12.1 [6.6] events per hour) (P<.002). Thirty-eight percent (10.6 events per hour) of the arousals in patients with beta-thalassemia and 25% (6.0 events per hour) of the arousals in patients with CDA-1 were induced by periodic limb movements during sleep. In the control group, most (98%) arousals were spontaneous and unrelated to any definable event. The multiple sleep latency test average was 7.8 minutes for patients with beta-thalassemia (n = 6) and 10.7 minutes for patients with CDA-1 (n = 8). Five patients with beta-thalassemia and 4 patients with CDA-1 underwent a second polysomnographic study on the next night to confirm reproducibility. There was no significant change in the total number or index of arousals and no difference in the severity of the periodic limb movements during sleep compared with the results of the first polysomnographic study. CONCLUSION: Children and adolescents with beta-thalassemia or CDA-1 have evidence of impaired sleep function that is partially due to periodic limb movements during sleep and arousals that result in objective diurnal sleepiness. PMID- 12742884 TI - Antiemetic use for acute gastroenteritis in children. AB - OBJECTIVES: To describe patterns of and factors associated with antiemetic use among a population-based sample of children with acute gastroenteritis; to determine if filling a prescription for an antiemetic is associated with a significant risk of adverse events; and to determine if filling a prescription for an antiemetic is associated with an increased risk of subsequent health care use. METHOD AND DESIGN: Retrospective cohort study of 20,222 children aged 1 month to 18 years, receiving Medicaid, who had a first diagnosis of gastroenteritis, diarrhea, or vomiting between January 1, 1998, and December 31, 1998. MAIN OUTCOME MEASURES: Presence of a claim for an antiemetic in the 3 days after the initial diagnosis of acute gastroenteritis, subsequent health care usage, and an adverse event within 14 days of the initial diagnosis. RESULTS: Parents of 1802 children (8.9%) with acute gastroenteritis had a prescription for an antiemetic filled within 3 days of the index visit. Factors associated with antiemetic prescription filling for children include older age, provider type (emergency physician, family physician, or general practitioner vs pediatrician), Spanish as the primary language, and rural residency. There was no difference in adverse events between children for whom an antiemetic prescription was filled and for those who did not have an antiemetic prescription filled (odds ratio, 0.68; 95% confidence interval, 0.31-1.46). No difference in risk of subsequent health care use was seen in children who had an antiemetic prescription filled and those who did not (incidence rate ratio, 1.04; 95% confidence interval, 0.94 1.16). CONCLUSION: Antiemetic use among children with acute gastroenteritis is common and adverse effects seem to be rare. PMID- 12742883 TI - Infant sleep position and associated health outcomes. AB - BACKGROUND: The incidence of sudden infant death syndrome has decreased in the United States as the percentage of infants sleeping prone has decreased, but persisting concerns about the safety of supine sleeping likely contribute to prone sleeping prevalence rates that remain higher than 10%. OBJECTIVE: To document health outcomes in infants aged 1 to 6 months in relation to sleep position. DESIGN: Prospective cohort study. SETTING: Massachusetts and Ohio, from February 21, 1995, to December 31, 1998. STUDY PARTICIPANTS: A total of 3733 infants with consistent sleep positions at ages 1, 3, and 6 months. MAIN OUTCOME MEASURES: Descriptive statistics and multiple logistic regression analysis relating sleep position at each follow-up age to symptoms in the prior week (fever, cough, wheezing, stuffy nose, trouble breathing or sleeping, diarrhea, vomiting, or spitting up) and outpatient visits in the prior month (ear infection, breathing problem, vomiting, spitting up, colic, seizure, accident, or injury). RESULTS: No symptoms or outpatient visits were significantly more common among infants sleeping on the side or supine than in infants sleeping prone, and 3 symptoms were less common: (1) fever at 1 month in infants sleeping in the supine (adjusted odds ratio [OR], 0.56; 95% confidence interval [CI], 0.34-0.93) and side positions (OR, 0.48; 95% CI, 0.28-0.82); (2) stuffy nose at 6 months in the supine (OR, 0.74; 95% CI, 0.61-0.89) and side positions (OR, 0.82; 95% CI, 0.68-0.99); and (3) trouble sleeping at 6 months in the supine (OR, 0.57; 95% CI, 0.44-0.73) and side positions (OR, 0.69; 95% CI, 0.53-0.89). Also, outpatient visits for ear infections were less common at 3 and 6 months in infants sleeping in the supine position (OR, 0.64; 95% CI, 0.46-0.88; and OR, 0.73; 95% CI, 0.58 0.92, respectively) and at 3 months in the side position (OR, 0.68; 95% CI, 0.49 0.96). CONCLUSIONS: No identified symptom or illness was significantly increased among nonprone sleepers during the first 6 months of life. These reassuring results may contribute to increased use of the supine position for infant sleeping. PMID- 12742885 TI - Using test dummy experiments to investigate pediatric injury risk in simulated short-distance falls. AB - BACKGROUND: Short-distance falls, such as from a bed, are often falsely reported scenarios in child abuse. In attempting to differentiate between abusive and nonabusive injury, knowledge of factors that affect injury risk in falls could prove useful. OBJECTIVES: To assess the biomechanics associated with simulated short-distance falls in children (one fall scenario, without attempting to maximize injury potential) and to investigate the effect of impact surface type on injury risk. METHODS: Repeatable fall experiments from bed height (0.68 m) onto different surfaces were conducted using an instrumented side-lying Hybrid II 3-year-old test dummy. Biomechanical measures assessed in falls included head acceleration, pelvis acceleration, femur loading, and head injury criteria. RESULTS: Fall dynamics resulted in the pelvis or legs making first contact. Biomechanical measures assessed in simulated bed falls were below known head injury criteria and lower extremity injury thresholds. The impact surface type had a significant effect on head injury risk and lower extremity loading. Playground foam proved to have the lowest associated injury risk of all the tested surfaces. CONCLUSIONS: The biomechanics of a child falling from a short distance, such as from a bed, were investigated using an experimental laboratory mock-up and an instrumented test dummy. Despite the impact surface having an effect on injury risk, rolling from a 0.68-m (27-in) horizontal surface from a side-lying posture presented low risk of contact-type head injury and leg injury on all tested impact surfaces. PMID- 12742886 TI - Early determination of ESR: how accurate is it? PMID- 12742887 TI - More attention to children's real health risks. PMID- 12742888 TI - Warts and duct tape--a good combo! PMID- 12742889 TI - Duct tape vs cryotherapy in the treatment of Verruca Vulgaris. PMID- 12742890 TI - Effective wart removal. PMID- 12742891 TI - Mechanism of wart disappearance: an alternative hypothesis. PMID- 12742893 TI - Cryotherapy vs duct tape. PMID- 12742892 TI - Duct tape occlusion for common warts: is effectiveness overestimated? PMID- 12742894 TI - Vulvovaginal candidiasis. PMID- 12742896 TI - Safety and efficacy of combination vaccines. PMID- 12742895 TI - Liver cancer in low and middle income countries. PMID- 12742898 TI - No-fault compensation systems. PMID- 12742897 TI - Management of bacterial meningitis in adults. PMID- 12742899 TI - Extracts from "concise clinical evidence". Commentary: treatment of diabetic retinopathy. PMID- 12742900 TI - WHO warns that death rate from SARS could reach 10%. PMID- 12742901 TI - Health committee questions benefit of foundation hospitals to patients. PMID- 12742902 TI - Food watchdog warns against high doses of vitamins and minerals. PMID- 12742903 TI - Complementary medicines industry in crisis after recall of 1546 products. PMID- 12742906 TI - Occupational injuries to NHS staff in England increase by a quarter. PMID- 12742909 TI - Child soldiers forgotten in Angola. PMID- 12742907 TI - Syrian doctor expelled from Hungary for allegedly supporting Hamas. PMID- 12742910 TI - Men should eat nine servings of fruit and vegetables a day. PMID- 12742912 TI - New guidance on ECT looks set to curb its use. PMID- 12742913 TI - FDA announces fast track approval of new drug for lung cancer. PMID- 12742915 TI - Advances in medical technology account for a third of the reduction in road traffic deaths. PMID- 12742917 TI - Containment of SARS depends on how it is handled in China. PMID- 12742916 TI - Toronto strives for normality after WHO lifts travel warning. PMID- 12742918 TI - WHO to create international human resources database on health care. PMID- 12742919 TI - Lundbeck broke advertising rules. PMID- 12742920 TI - Co-proxamol and suicide: a study of national mortality statistics and local non fatal self poisonings. AB - OBJECTIVES: To examine the incidence of suicides due to co-proxamol compared with tricyclic antidepressants and paracetamol, and to compare fatality rates for self poisonings with these drugs. DESIGN: Analysis of routinely collected national and local data on suicides and self poisonings. SETTING: Records of suicides in England and Wales 1997-9; non-fatal self poisonings in Oxford District 1997-9. DATA SOURCES: Office for National Statistics and Oxford monitoring system for attempted suicide. MAIN OUTCOME MEASURES: Incidence of suicides with co-proxamol or tricyclic antidepressants or paracetamol. Ratios of fatal to non-fatal self poisonings. RESULTS: Co-proxamol alone accounted for 5% of all suicides. Of 4162 drug related suicides, 18% (766) involved co-proxamol alone, 22% (927) tricyclic antidepressants alone, and 9% (368) paracetamol alone. A higher proportion of suicides in the 10-24 year age group were due to co-proxamol than in the other age groups. The odds of dying after overdose with co-proxamol was 2.3 times (95% confidence interval 2.1 to 2.5) that for tricyclic antidepressants and 28.1 times (24.9 to 32.9) that for paracetamol. CONCLUSIONS: Self poisoning with co-proxamol is particularly dangerous and contributes substantially to drug related suicides. Restricting availability of co-proxamol could have an important role in suicide prevention. PMID- 12742921 TI - Association between antidepressant prescribing and suicide in Australia, 1991 2000: trend analysis. AB - OBJECTIVE: To examine the association between trends in antidepressant prescribing and suicide rates in Australia for 1991-2000. DESIGN: Analysis of databases of suicide and rates of antidepressant prescribing according to age and sex. SETTING: Australian Bureau of Statistics data, sales data from the Australian pharmaceutical industry, prescribing data in general practice. SUBJECTS: Men and women aged 15 years and over in 10 year age groups. MAIN OUTCOME MEASURES: Trends in suicide rates and trends in antidepressant prescribing. Association measured by Spearman's rank correlations. RESULTS: While overall national rates of suicide did not fall significantly, incidence decreased in older men and women and increased in younger adults. In both men (r(s)=-0.91; P<0.01) and women (r(s)=-0.76; P<0.05) the higher the exposure to antidepressants the larger the decline in rate of suicide. CONCLUSIONS: Changes in suicide rates and exposure to antidepressants in Australia for 1991-2000 are significantly associated. This effect is most apparent in older age groups, in which rates of suicide decreased substantially in association with exposure to antidepressants. The increase in antidepressant prescribing may be a proxy marker for improved overall management of depression. If so, increased prescribing of selective serotonin reuptake inhibitors in general practice may have produced a quantifiable benefit in population mental health. PMID- 12742922 TI - Preregistration house officers' views on whether their experience at medical school prepared them well for their jobs: national questionnaire survey. PMID- 12742923 TI - Prospective randomised controlled trial of laparoscopic versus open inguinal hernia mesh repair: five year follow up. PMID- 12742925 TI - 10-minute consultation: adverse drug event. PMID- 12742924 TI - Efficacy and tolerability of selective serotonin reuptake inhibitors compared with tricyclic antidepressants in depression treated in primary care: systematic review and meta-analysis. AB - OBJECTIVE: To compare the efficacy and tolerability of tricyclic antidepressants with selective serotonin reuptake inhibitors in depression in primary care. DESIGN: Systematic review and meta-analysis of randomised controlled trials. DATA SOURCES: Register of the Cochrane Collaboration's depression, anxiety, and neurosis group. Reference lists of initial studies and other relevant review papers. Selected authors and experts. SELECTION OF STUDIES: Studies had to meet minimum requirements on: adequacy of sample size, adequate allocation concealment, clear description of treatment, representative source of subjects, use of diagnostic criteria or clear specification of inclusion criteria, details regarding number and reasons for withdrawal by group, and outcome measures described clearly or use of validated instruments. MAIN OUTCOME MEASURES: Standardised mean difference of final mean depression scores and relative risk of response when using the clinical global impression score. Relative risk of withdrawing from treatment at any time, and the number withdrawing due to side effects. RESULTS: 11 studies (2951 participants) compared a selective serotonin reuptake inhibitor with a tricyclic antidepressant. Efficacy between selective serotonin reuptake inhibitors and tricyclics did not differ significantly (standardised weighted mean difference, fixed effects 0.07, 95% confidence interval -0.02 to 0.15; z=1.59, P<0.11). Significantly more patients receiving a tricyclic withdrew from treatment (relative risk 0.78, 95% confidence interval 0.68 to 0.90; z=3.37, P<0.0007) and withdrew specifically because of side effects (0.73, 0.60 to 0.88; z=3.24, P<0.001). Most studies included were small and supported by commercial funding. Many studies were of low methodological quality or did not present adequate data for analysis, or both, and were of short duration, typically six to eight weeks. CONCLUSION: The evidence on the relative efficacy of selective serotonin reuptake inhibitors and tricyclic antidepressants in primary care is sparse and of variable quality. The study setting is likely to be an important factor in assessing the efficacy and tolerability of treatment with antidepressant drugs. PMID- 12742926 TI - Preregistration house officers in general practice: review of evidence. AB - OBJECTIVES: To examine the strengths and weaknesses of the national and local schemes for preregistration house officers to spend four months in general practice, to identify any added value from such placements, and to examine the impact on career choices. DESIGN: Review of all studies that reported on placements of preregistration house officers in general practice. SETTING: 19 accounts of preregistration house officers' experience in general practice, ranging from single case reports to a national evaluation study, in a variety of locations in Scotland and England. PARTICIPANTS: Views of 180 preregistration house officers, 45 general practitioner trainers, and 105 consultant trainers. MAIN OUTCOME MEASURES: Main findings or themes weighted according to number of studies reporting them and weighted for sample size. RESULTS: The studies were unanimous about the educational benefits of the placements. The additional learning included communication skills, social and psychological factors in illness, patient centred consultations, broadening of knowledge base, and dealing with uncertainty about diagnosis and referral. CONCLUSIONS: Despite the reported benefits and recommendations of the scheme, it is not expanding. General practitioner trainers reported additional supervision that was unremunerated. The reforms of the senior house officer grade may resolve this problem by offering the placements to senior house officers, who require less supervision. PMID- 12742927 TI - Extracts from "concise clinical evidence". Diabetic retinopathy. PMID- 12742929 TI - Pathophysiology and investigation of coronary artery disease. PMID- 12742928 TI - Zosteriform metastasis from melanoma. PMID- 12742930 TI - Giving citizens a voice in healthcare policy in Canada. PMID- 12742931 TI - Ten ways to improve information technology in the NHS. Users, not geeks, should design interface. PMID- 12742932 TI - Closing the digital divide. Reality may not be so rosy. PMID- 12742933 TI - Folate and risk of cardiovascular disease. Study results were misinterpreted. PMID- 12742934 TI - Chronic cough. Article is not consistent with WHO initiative on rhinitis and asthma. PMID- 12742935 TI - Medical experts and the criminal courts. Meaningful audit could be difficult to attain. PMID- 12742936 TI - Leishmaniasis also occurs in Mediterranean countries. PMID- 12742937 TI - Encouraging insensitivity in doctors is not an option. PMID- 12742938 TI - Cleft lip and palate services still need to be improved. PMID- 12742939 TI - Involving community may be way forward post-Shipman. PMID- 12742940 TI - Difficulties in giving fully informed consent. PMID- 12742941 TI - Parents are not homogeneous. PMID- 12742942 TI - Senior medical staff need to be included in severe asthma. PMID- 12742943 TI - A career in rheumatology. PMID- 12742947 TI - We are the gatekeepers. PMID- 12742948 TI - High success with nonoperative management of blunt hepatic trauma: the liver is a sturdy organ. AB - HYPOTHESIS: Nonoperative management of liver injuries (NOMLI) is highly successful and rarely leads to adverse events. DESIGN: Prospective observational study. SETTING: High-volume academic level I trauma center. PATIENTS: For 26 months, 78 consecutive unselected patients with liver injuries were followed up prospectively. In the absence of hemodynamic instability or signs of hollow visceral trauma, NOMLI was offered irrespective of the magnitude of the liver injury. Main Outcome Measure Failure of NOMLI, defined as a laparotomy after an initial decision to treat the patient nonoperatively. RESULTS: Of the 78 patients, 23 (29%) were operated on immediately, but only 12 (15%) for bleeding from the liver. All 12 patients required packing in addition to other maneuvers (hepatorrhaphy [n = 8], resection [n = 4], and liver isolation [n = 1]). Of the remaining 55 patients selected for NOMLI, the method failed in 8 for reasons unrelated to the liver injury: 2 underwent a splenectomy, 1 underwent a nephrectomy, 1 had a small-bowel repair, 1 underwent abdominal decompression for abdominal compartment syndrome, and 3 underwent a nontherapeutic laparotomy. The success rate of NOMLI was 85% (47 of 55 patients), but the liver-specific success rate was 100%. Compared with those in whom NOMLI was successful, patients in whom it failed had a higher Injury Severity Score and underwent more blood transfusions, but they had similar liver injury grades. In total, 66 (85%) of liver injuries did not bleed significantly. No adverse events were attributed to NOMLI. CONCLUSIONS: Nonoperative management of liver injuries is safe and effective regardless of the grade of liver injury. Failure of NOMLI is caused by associated abdominal injuries and not the liver. Fluid and blood requirements, the degree of injury severity, and the presence of other abdominal organ injuries may help predict failure. PMID- 12742950 TI - The selective use of sentinel node biopsy in ductal carcinoma in situ. PMID- 12742949 TI - Relapse and morbidity in patients undergoing sentinel lymph node biopsy alone or with axillary dissection for breast cancer. AB - HYPOTHESIS: Axillary relapse in node-negative patients staged with sentinel lymph node (SLN) biopsy alone is no more frequent than in patients treated with standard axillary dissection. Morbidity is less for patients who had SLN biopsy. DESIGN, SETTING, AND PATIENTS: Between October 14, 1997, and August 31, 2001, 1253 consecutive women with primary invasive breast cancer were prospectively entered into an SLN biopsy database. Completion axillary dissection was performed in 164 patients after SLN biopsy as part of a training protocol. INTERVENTIONS: Patients were contacted by questionnaire or telephone to determine breast cancer relapse; presence of arm lymphedema, arm pain, axillary infection, or seroma formation; and tumor recurrence or death. MAIN OUTCOME MEASURES: chi2 or Fisher exact tests and Wilcoxon rank sum tests were used to analyze categorical and continuous variables. Logistic regression was used to analyze morbidity. RESULTS: Of 1253 women, 894 (71%) were node negative by SLN biopsy alone (n = 730 [82%]) or SLN biopsy and completion axillary dissection (n = 164 [18%]). Questionnaires were completed by 776 patients (87%). Mean +/- SD follow-up was 2.4 +/- 0.9 years. Patients with axillary dissections reported a significantly higher occurrence of arm lymphedema (34%), arm pain (38%), seroma formation (24%), and infection (9%) vs SLN biopsy-only patients (6%, 14%, 7%, and 3%, respectively). One axillary relapse (0.1%) occurred during follow-up of 685 women who underwent SLN biopsy only. CONCLUSIONS: With intermediate-term follow-up, there was 1 axillary recurrence in 685 SLN node-negative women, supporting use of SLN biopsy as an accurate method for staging breast cancer. Biopsy of the SLN was associated with significantly less morbidity than completion axillary dissection. PMID- 12742951 TI - Improved outcome after extended gastric myotomy for achalasia. AB - HYPOTHESIS: There is general agreement that a Heller myotomy should extend 6 to 7 cm above the gastroesophageal junction. Results of most previous studies have recommended that the myotomy extend 1 to 1.5 cm below the gastroesophageal junction. We speculated that the effectiveness of the operation could be improved if a longer, 3-cm myotomy was carried out below the gastroesophageal junction, as it would more completely obliterate the lower esophageal sphincter. We, therefore, changed our technique in 1998. Concurrently, we converted from a Dor fundoplication to a Toupet fundoplication. This study analyzes the results of our new strategy. DESIGN: A case series using a prospectively maintained database. SETTING: Tertiary referral center. PATIENTS: One hundred ten consecutive patients with achalasia undergoing laparoscopic Heller myotomy. INTERVENTION: We analyzed the course of 52 patients treated with a standard laparoscopic esophagogastric myotomy (1.5 cm in the stomach) and a Dor fundoplication between September 1, 1994, and August 31, 1998, and 58 treated with an extended gastric myotomy (3 cm below the gastroesophageal junction) and a Toupet fundoplication between September 1, 1998, and August 31, 2001. MAIN OUTCOME MEASURES: Esophageal function testing (esophageal manometry and 24-hour pH monitoring), symptom questionnaire (frequency and severity), and postoperative interventions required. RESULTS: Postoperatively the lower esophageal sphincter pressure was significantly lower after extended gastric myotomy and a Toupet fundoplication vs standard myotomy and a Dor fundoplication (9.5 vs 15.8 mm Hg). Dysphagia was both less frequent (1.2 vs 2.1) and less severe (visual analog scale, 3.2 vs 5.3) after extended gastric myotomy and Toupet fundoplication. In the standard laparoscopic esophagogastric myotomy and a Dor fundoplication group, 9 patients (17%) had recurrent, severe dysphagia, which was treated by dilation in 5 patients and by reoperation in 4 patients. In the extended gastric myotomy and Toupet fundoplication group, 2 patients (3%) developed recurrent dysphagia that resolved with dilatation. There were no reoperations in the extended gastric myotomy and Toupet fundoplication group. No difference was noted in the frequency of heartburn (1.3 vs 1.7), regurgitation (0.3 vs 0.8), and chest pain (0.3 vs 0.6), nor was there a difference between the 2 groups in proximal (1.7% vs 2.3%) and distal (6.0% vs 5.9%) esophageal acid exposure. CONCLUSION: An extended gastric myotomy (3 cm) more effectively disrupts the lower esophageal sphincter, thus improving the results of surgical therapy for achalasia for dysphagia without increasing the rate of abnormal gastroesophageal reflux provided that a Toupet fundoplication is added. PMID- 12742952 TI - Long-term follow-up of the modified Delorme procedure for rectal prolapse. AB - HYPOTHESIS: The modified Delorme operation is a safe, effective, and durable treatment for complete rectal prolapse. DESIGN: Retrospective analysis of outcomes in adult patients undergoing the modified Delorme operation. SETTING: Community-based tertiary referral center with a 5-year general surgery residency program. PATIENTS: A total of 52 consecutive patients undergoing surgery for the treatment of complete rectal prolapse during the 26-year period ending December 2001. INTERVENTIONS: Modified Delorme operation. Main Outcomes Measured Method of anesthesia, morbidity, mortality, recurrence rates, length of follow-up, and incontinence. RESULTS: In the 52 patients, the mean length of prolapse was 8.2 cm. The mean operating time was 75 minutes. Forty-five patients were administered general anesthesia, 4 were administered spinal anesthesia, and 3 were administered local anesthesia. The mean postoperative stay was 4.9 days for 1975 through 2001 and 2.8 days for 1990 through 2001. No patients died as a result of the procedure. Patients were followed up for 61.4 months. Major medical comorbidities occurred in 40 patients. Preoperative incontinence was present in 12 patients, 10 of whom improved after the procedure, and postoperative incontinence in 8. The recurrent postoperative prolapse rate at 5 years was 6% (3/52) and the recurrent postoperative prolapse rate to the end of the study was 10% (5/52). Two patients (4%) had complications that required operative intervention in the postoperative period. CONCLUSIONS: The modified Delorme operation is a safe and effective surgical treatment for complete rectal prolapse. The risk of recurrent prolapse is low, and the procedure may be safely performed in patients with significant medical comorbidities. PMID- 12742953 TI - Assessment of volume of hemorrhage and outcome from pelvic fracture. AB - HYPOTHESIS: Measurement of pelvic hemorrhage on computed tomographic (CT) scans can estimate the pelvic fracture component of total patient blood loss and predict the need for angiography. DESIGN: Retrospective cohort study. SETTING: Large level 1 trauma center. PATIENTS: We examined data from 759 consecutive, nonreferral blunt trauma patients who sustained pelvic fracture. MAIN OUTCOME MEASURES: Pelvic-fracture-specific outcomes included estimation of extraperitoneal pelvic hemorrhage volume from emergency department CT scans and determination of arterial injury from angiograms. General patient outcomes determined from medical record review included transfusion requirement, estimated blood loss, and mortality. Subanalysis was performed on subjects with only pelvic fracture as a source of major hemorrhage (derived from discharge International Classification of Diseases, Ninth Revision, Clinical Modification codes). RESULTS: Overall mortality was 96 (13%) of 759 patients. Blood transfusion was given to 418 (55%) patients, and 258 (34%) received 6 or more units in the first 72 hours. Pelvic-fracture-related hemorrhage averaged 149 mL (range, 0-1423 mL). Angiography was performed on 163 patients, of whom 113 had arterial injury. Higher pelvic hemorrhage volumes on CT scans were seen in subjects with pelvic arterial injury demonstrated on angiograms (P<.001). In subjects without another source of major hemorrhage, pelvic CT hemorrhage volumes were strongly associated with transfusion requirement (P<.001). Subjects with large pelvic hemorrhage volumes (>500 mL) were more likely to have pelvic arterial injury (risk ratio, 4.8; 95% confidence interval, 3.0-7.8; P<.001) and require large-volume (>/=6 U) transfusions (risk ratio, 4.7; 95% confidence interval, 1.8-12.3; P<.001) than patients with smaller pelvic hemorrhage volumes. CONCLUSION: Pelvic hemorrhage volumes derived from pelvic CT scans were predictors of the need for pelvic arteriography and transfusions. PMID- 12742954 TI - The efficacy of percutaneous transluminal angioplasty in the treatment of infrainguinal vein bypass graft stenosis. AB - HYPOTHESIS: Percutaneous angioplasty would provide a durable alternative to surgical revision in the treatment of infrainguinal vein graft stenosis. DESIGN: Outcome analysis of the results of percutaneous angioplasty of infrainguinal vein graft stenosis. SETTING: Academic vascular surgical practice in a university affiliated community hospital. PARTICIPANTS: All patients undergoing percutaneous intervention for infrainguinal vein graft stenosis from January 1, 1995, to May 31, 2002, were enrolled in the study. INTERVENTIONS: Lower extremity arterial reconstruction was performed by one of us. Proximal and distal sites of graft placement were identified, as well as the conduit used. Percutaneous angioplasty was performed on grafts by 1 of 4 interventional radiologists. Criteria for intervention and the anatomic location of intervention were noted. Morbidity from percutaneous intervention was also determined. MAIN OUTCOME MEASURES: Success and durability of percutaneous angioplasty were determined by clinical follow-up, duplex surveillance, and arteriography. Failure was defined as duplex ultrasonographic or arteriographic documentation of stenosis of 75% or greater. Kaplan-Meier life table analysis was applied to all grafts in the study. RESULTS: Ninety-four patients with 101 grafts were included in the study. Nearly 35% of angioplasties had failed at 6 months, 53.6% had failed at 12 months, 60.6% had failed at 24 months, and 75.1% had failed at 36 months. Comorbid disease, use of anticoagulant medications, criteria for intervention, or anatomic location of percutaneous intervention did not affect patency. Eight angioplasties (7.9%) were associated with significant complications. CONCLUSIONS: Percutaneous angioplasty does not provide a durable solution to the problem of infrainguinal vein graft stenosis. Because of the high rate of complications, its routine use cannot be advocated. PMID- 12742955 TI - Symptoms are a poor indicator of reflux status after fundoplication for gastroesophageal reflux disease: role of esophageal functions tests. AB - BACKGROUND: If a patient develops foregut symptoms after a fundoplication, it is assumed that the operation has failed, and acid-reducing medications are often prescribed. Esophageal function tests (manometry and pH monitoring) are seldom performed early in the management of these patients. HYPOTHESIS: In patients who are symptomatic after fundoplication for gastroesophageal reflux disease, a symptom-based diagnosis is not accurate, and esophageal function tests should be performed routinely before starting acid-reducing medications. DESIGN: Prospective study. SETTING: University hospital. PATIENTS AND METHODS: One hundred twenty-four patients who developed foregut symptoms after laparoscopic fundoplication (average, 17 months postoperatively) underwent esophageal manometry and pH monitoring. Sixty-two patients (50%) were taking acid-reducing medications. MAIN OUTCOME MEASURES: Postoperative symptoms, use of antireflux medications, grade of esophagitis, esophageal motility, and DeMeester scores. RESULTS: Seventy-six (61%) of the 124 patients had normal esophageal acid exposure, while the acid exposure was abnormal in 48 patients (39%). Only 20 (32%) of the 62 patients who were taking acid-reducing medications had reflux postoperatively. Regurgitation was the only symptom that predicted abnormal reflux. CONCLUSIONS: These results show that (1) symptoms were due to reflux in 39% of patients only; (2) with the exception of regurgitation, symptoms were an unreliable index of the presence of reflux; and (3) 68% of patients who were taking acid-reducing medications postoperatively had a normal reflux status. Esophageal function tests should be performed early in the evaluation of patients after fundoplication to avoid improper and costly medical therapy. PMID- 12742956 TI - Roux-en-Y gastric bypass leak complications. AB - HYPOTHESIS: Enteric leakage is a significant complication of the Roux-en-Y gastric bypass (RYGB) procedure that can be treated successfully. DESIGN: Retrospective study of 400 consecutive RYGB patients from 1999-2002. SETTING: Community hospital with a university surgical residency. PATIENTS: Hospital records of 400 morbidly obese patients who underwent gastric bypass surgery were reviewed. MAIN OUTCOME MEASURES: Time of discovery of leak, location of leak, treatment, hospital stay, and mortality. RESULTS: Twenty-one patients (5.25%) developed leaks. The mean body mass index (calculated as weight in kilograms divided by the square of height in meters) was 54.2. Thirteen patients were noted to develop a leak at the gastrojejunal anastomosis, with an average time to diagnosis of 7.0 days. Five of these patients underwent reexploration, and 8 were successfully treated with percutaneous drainage alone. Four patients developed leaks at the jejunojejunal anastomosis (mean time to diagnosis, 2.0 days). All of these patients required exploration, and 2 patients died. Four patients were noted to have leaks in other areas (average time to diagnosis, 3.5 days). Two patients were treated with drainage, and 2 underwent exploration. The average hospital stay of all patients was 33 days. CONCLUSIONS: Enteric leakage is a significant complication of the RYGB. Patients who are suspected of having an enteric leak because of signs of sepsis or hemodynamic instability require emergent exploration. Leaks that are more insidious may be treated successfully with percutaneous drainage. Aggressive exploration of patients who appear to be septic, and percutaneous drainage of insidiously developing leaks may decrease patients' morbidity and mortality. PMID- 12742958 TI - Laparoscopic cholecystectomy for elderly patients: gold standard for golden years? AB - HYPOTHESIS: Laparoscopic cholecystectomy (LC) has known physiological benefits and positive socioeconomic effects over the open procedure. Although recent studies have questioned the technique's efficacy in elderly patients (>65 years), we hypothesize that LC is safe and efficacious in that patient group. METHODS: Five thousand eight hundred eighty-four consecutive patients (mean age, 40 years; 26% male) underwent an attempted LC (conversion rate, 5.2%) from 1991 to 2001 at a teaching institution. Of these, 395 patients (6.7%) were older than 65 years. Analysis included patient age, sex, American Society of Anesthesiologists classification, conversion rate, morbidity, mortality, and assessment of results over time. RESULTS: Elderly patients were predominantly male (64%). Septuagenarians had a 40% incidence of complicated gallstone disease, such as acute cholecystitis, choledocholithiasis, or biliary pancreatitis, and octogenarians had a 55% incidence. Overall mortality was 1.4%. The conversion rate was 17% for the first 5 years of the study period and 7% for the second half. The conversion rate was 22% for patients with complicated disease and 2.5% for patients with chronic cholecystitis. Average hospital stay decreased from 10.2 days to 4.6 days during the first and second half of the study period, respectively. CONCLUSIONS: The results of LC in patients aged 65 to 69 years are comparable with those previously reported in younger patients. Patients older than 70 years had a 2-fold increase in complicated biliary tract disease and conversion rates, but a low mortality rate (2%) compared with results of other authors (12%), despite an increase in American Society of Anesthesiologists classification. Increased technical experience with LC favorably affected outcomes over time. Early diagnosis and treatment prior to onset of complications are necessary for further improvement in the outcomes of elderly patients undergoing LC. PMID- 12742957 TI - The Millikan modified mesh-plug hernioplasty. AB - HYPOTHESIS: A modified technique for mesh-plug hernioplasty is a safe and efficacious option for primary unilateral inguinal herniorrhaphy. DESIGN: Prospective analysis of 1056 patients who underwent primary unilateral inguinal hernioplasty. SETTING: A private university medical center. PATIENTS: One thousand twenty-five men and 31 women (mean age, 49 years) with primary unilateral inguinal hernias that were surgically repaired between May 1, 1997, and November 1, 2001. INTERVENTION: We performed a modified technique using a mesh plug and local anesthesia with intravenous sedation. The modified technique consisted of placing the mesh plug into the preperitoneal space and suture fixation of the plug using the inner petals. MAIN OUTCOME MEASURES: Surgical morbidity, hernia recurrence, postoperative pain medication used, and return to normal activities. RESULTS: We included 642 indirect and 414 direct hernias. Mean operative time was 25 minutes; mean recovery room time, 45 minutes. All procedures were performed as outpatient surgery. One thousand thirteen patients (95.9%) returned to normal activities within 3 days. All manual laborers returned to work on postoperative day 14. Only 169 patients (16.0%) required prescription pain medication. At 1-year follow-up, 1045 patients (99.0%) have been examined, and 1 recurrence (0.1%) has been detected. No mesh infection has occurred, and 19 hematomas spontaneously resolved. Five patients (0.5%) required treatment for persistent postoperative pain. CONCLUSIONS: The modified mesh-plug hernioplasty uses a minimum of medical resources and is associated with a small amount of postoperative pain and an early return to normal activities and manual labor with a minimal documented early recurrence rate. The Millikan modified mesh-plug hernioplasty should be adopted as the gold standard for unilateral primary inguinal hernioplasty. PMID- 12742959 TI - Casting-type calcifications with invasion and high-grade ductal carcinoma in situ: a more aggressive disease? AB - HYPOTHESIS: Women with breast cancer who have casting-type microcalcifications associated with multifocal invasion and extensive ductal carcinoma in situ (DCIS) form a subset of patients with a poor prognosis. Our study aims to identify the mammographic and pathologic features of this group. DESIGN: Women with casting type microcalcifications, multifocal invasion, and extensive DCIS were identified from our tumor board registry. Mammographic features, tumor characteristics, treatment, and survival rates were evaluated. Invasive tumors were limited to 14 mm or smaller. SETTING: University medical teaching hospital and breast cancer specialty clinic. RESULTS: Of the 984 patients with breast cancer treated at our center, 15 patients were identified who had extensive casting-type calcifications and DCIS. Twelve of these patients also had multifocal invasive breast cancer. All had casting-type microcalcifications occupying more than 1 breast quadrant. All but 1 of the patients were treated using mastectomy with sentinel node biopsy or axillary node dissection. All but 1 patient had extensive grade 3 DCIS. Invasive tumors were negative for estrogen receptor and progesterone receptor expression in half of the patients, and 60% were positive for the HER-2-neu receptor. Positive axillary lymph nodes were found in 33% of patients, and 75% received adjuvant chemotherapy. After a median follow-up period of 20.5 months (range, 6-72 months), 1 patient had died and 1 had distant metastases. Of the 3 patients who had DCIS without invasion, 1 experienced a recurrence with infiltrating ductal carcinoma. CONCLUSIONS: In women with small multifocal breast cancers with extensive casting calcifications and DCIS, the incidence of positive lymph nodes was 33%, with a tendency for poor tumor markers. These women appear to be at substantial risk for systemic disease; lymph node sampling and adjuvant systemic therapy are recommended. PMID- 12742960 TI - Predictors of complication and suboptimal weight loss after laparoscopic Roux-en Y gastric bypass: a series of 188 patients. AB - HYPOTHESIS: An analysis of patients undergoing laparoscopic Roux-en-Y gastric bypass (RYGB) may identify factors predictive of complication and of suboptimal weight loss. DESIGN: Inception cohort. SETTING: Metropolitan university hospital. PATIENTS: One hundred eighty-eight consecutive patients with severe obesity who met National Institutes of Health consensus guidelines for bariatric surgery. INTERVENTIONS: Laparoscopic RYGB. MAIN OUTCOME MEASURES: Complications requiring therapeutic intervention and percentage of excess body weight lost at 1 year after surgery. RESULTS: Of the 188 patients who underwent laparoscopic RYGB, 50 (26.6%) developed complications that required an invasive therapeutic intervention, including 2 deaths. The average follow-up was 351 days (range, 89 1019 days). Multivariate analysis by stepwise logistic regression identified surgeon experience, sleep apnea (P =.003; odds ratio, 3.0; 95% confidence interval, 1.3-7.1), and hypertension (P =.07; odds ratio, 2.0; 95% confidence interval, 1.0-4.0) as predictors of complications. The most common complication requiring therapeutic intervention was stricture at the gastrojejunal anastomosis, occurring in 27 patients (14.4%). Of the 115 patients who underwent surgery more than 1 year previously, 1-year follow-up data were available for 93 (81%). The body mass index (weight in kilograms divided by the square of height in meters) decreased from 53 +/- 8 preoperatively to 35 +/- 6 at 1 year. The mean +/- SD percentage of excess body weight lost at 1 year was 61% +/- 14%. Diabetes mellitus was negatively correlated with percentage of excess body weight lost at 1 year (P =.06). CONCLUSIONS: Surgeon experience, sleep apnea, and hypertension are associated with complications after laparoscopic RYGB. Diabetes mellitus may be associated with poorer postoperative weight loss. PMID- 12742961 TI - Outcome of right hepatectomies in patients older than 70 years. AB - HYPOTHESIS: The increasing number of elderly patients undergoing liver resections mandates updating of clinical outcomes on this specific population. DESIGN: Case series. SETTING: A tertiary care teaching hospital. PATIENTS: Twenty-three patients older than 70 years who underwent right hepatectomies (including 7 extended right hepatectomies) between January 1, 1995, and October 31, 2001 (group 1) and 99 patients younger than 70 years who underwent 64 right hepatectomies and 35 extended right hepatectomies during the same period (group 2) were included for a total sample population of 122. MAIN OUTCOME MEASURES: Preoperative clinicopathological features, intraoperative factors, in-hospital mortality, postoperative complications, intensive care unit requirement, hospital stay, and course of main biochemical liver function test results of groups 1 and 2 were analyzed and compared. RESULTS: The 2 groups were similar for indications for surgery and the presence of underlying liver disease. Group 1 had a higher incidence of associated pulmonary diseases (21.7% vs 5%, P =.02) and patients with an American Society of Anesthesiologists score of III (ie, a patient with severe systemic disease limiting activity, but not incapacitating) (56.5% vs 26.3% of cases, P =.01). There were no differences in intraoperative requirement of packed red blood cells and in operation time. There were no in-hospital deaths in group 1; there were 2 deaths (2%) in group 2. Nine patients (39.1%) in group 1 and 32 patients (32.3%) in group 2 experienced postoperative complications (P =.53), of whom, respectively, 5 (21.7%) and 17 (17.2%) developed transient liver dysfunction (P =.56), and 4 (17.4%) and 5 (5.1%) required a supplementary intesive care unit stay (P =.06). The postoperative stay (mean [SD], 16 [14] days vs 13 [9] days, P =.88) and peak values of the aminotransferase level, total serum bilirubin level, and prothrombin time were similar in the 2 groups. The timing of the peak value of the total serum bilirubin level (mean [SD], 4.1 [4.8] days vs 2.5 [2.5] days, P =.28) and its period of normalization (mean [SD], 9.4 [10.8] days vs 6.7 [5.1] days, P =.67) were also similar for both groups. For patients with malignancies, the 3-year survival rate was 64.2% in group 1 and 53.9% in group 2 (P =.53). CONCLUSION: Being older than 70 years should not be a contraindication for major hepatectomies, provided that liver cirrhosis and severe associated medical conditions are ruled out during the preoperative evaluation. PMID- 12742962 TI - Perineal dissection of synchronous abdominoperineal resection of the rectum: an anatomical description. PMID- 12742963 TI - Image of the month. Foramen of Winslow hernia. PMID- 12742964 TI - Incidents in surgical history. PMID- 12742965 TI - Mercedes speed. PMID- 12742966 TI - Temporary abdominal coverage and abdominal compartment syndrome. PMID- 12742967 TI - Percutaneous treatment of saphenous vein bypass graft obstructions: a continuing obstinate problem. PMID- 12742969 TI - Images in cardiovascular medicine. Real-time perfusion echocardiography of an intracardiac mass. PMID- 12742968 TI - New frontiers in cardiology: drug-eluting stents: Part II. PMID- 12742970 TI - Cardiology patient page. Beta-adrenergic blockers. PMID- 12742971 TI - Images in cardiovascular medicine. Tako-tsubo--like transient left ventricular dysfunction. PMID- 12742972 TI - The Brugada numbers. PMID- 12742973 TI - C-reactive protein and electron beam tomography. PMID- 12742974 TI - Cardiac troponin I predicts short-term mortality in vascular surgery patients. PMID- 12742975 TI - Myocardial perfusion in patients with transposition of the great arteries after arterial switch operation. PMID- 12742976 TI - Effect of transdermal estradiol and oral conjugated equine estrogen on C-reactive protein in retinoid-placebo trial in healthy women. PMID- 12742978 TI - Use of rapamycin slows progression of cardiac transplantation vasculopathy. AB - BACKGROUND: Cardiac transplantation vasculopathy is the leading cause of late death in heart transplantation recipients. Rapamycin is an immunosuppressant drug with potent antiproliferative and antimigratory effects. We investigated whether rapamycin could prevent progression of graft vasculopathy in 46 patients (age, 54+/-10 years; 4.3+/-2.3 years after transplantation) with severe disease. METHODS AND RESULTS: At annual cardiac catheterization, patients were randomly assigned to treatment with rapamycin (n=22) versus continued current immunosuppression (n=24). Clinical characteristics including recipient age and sex, underlying cause of congestive heart failure, donor age and sex, and ischemic time were recorded. Cardiac catheterization was graded with the use of a semiquantitative scale and repeated annually. Clinically significant adverse events were defined as death, need for angioplasty or bypass surgery, myocardial infarction, and a >25% worsening of the catheterization score. These events were monitored as primary study end points. Anti-HLA class I and II antibody production and lymphocyte growth assays were measured with each biopsy. Patients selected for rapamycin had azathioprine or mycophenolate mofetil discontinued and were given rapamycin. Outcomes were compared by means of log-rank analysis. There were no significant differences in baseline characteristics. Duration of follow up was comparable (rapamycin, 689+/-261; control, 630+/-207 days; NS). In the rapamycin group, 3 patients reached primary end points versus 14 patients in the control group (P<0.001). There was no difference in baseline or subsequent anti HLA class I or II antibody production. CONCLUSIONS: In this patient cohort with cardiac vasculopathy, treatment with rapamycin slowed disease progression probably by its antiproliferative and antimigratory effects. PMID- 12742979 TI - Vasopressin V2-receptor blockade with tolvaptan in patients with chronic heart failure: results from a double-blind, randomized trial. AB - BACKGROUND: In this study, we evaluated the effects of tolvaptan (OPC-41061), a novel, oral, nonpeptide vasopressin V2-receptor antagonist in patients with chronic heart failure (CHF). METHODS AND RESULTS: This was a double-blind study investigating the effects of three doses of tolvaptan and placebo in patients with CHF. After a run-in period, 254 patients were randomly assigned to placebo (n=63) or tolvaptan [30 mg (n=64), 45 mg (n=64), or 60 mg (n=63)] once daily for 25 days. Patients were not fluid-restricted and were maintained on stable doses of furosemide. At day 1, when compared with baseline, a decrease in body weight of -0.79+/-0.99, -0.96+/-0.93, and -0.84+/-0.02 kg was observed in the 30-, 45-, and 60-mg tolvaptan groups, respectively, and a body weight increase of +0.32+/ 0.46 kg in the placebo group (P<0.001 for all treatment groups versus placebo). Although the initial decrease in body weight was maintained during the study, no further reduction was observed beyond the first day. An increase in urine volume was observed with tolvaptan when compared with placebo (3.9+/-0.6, 4.2+/-0.9, 4.6+/-0.4, and 2.3+/-0.2 L/24 hours at day 1 for 30-, 45-, and 60-mg tolvaptan groups, and placebo, respectively; P<0.001). A decrease in edema and a normalization of serum sodium in patients with hyponatremia were observed in the tolvaptan group but not in the placebo group. No significant changes in heart rate, blood pressure, serum potassium, or renal function were observed. CONCLUSIONS: In patients with CHF, tolvaptan was well tolerated; it reduced body weight and edema and normalized serum sodium in the hyponatremic patients. PMID- 12742980 TI - Incomplete resolution of ST-segment elevation is a marker of transient microcirculatory dysfunction after stenting for acute myocardial infarction. AB - BACKGROUND: Incomplete ST-segment resolution (STR) after successful primary angioplasty for acute myocardial infarction (AMI) is associated with a poor prognosis. We used intracoronary Doppler velocimetry to investigate whether incomplete STR after primary angioplasty is a marker of severe microcirculatory dysfunction. METHODS AND RESULTS: Fifty patients with < or =12-hour AMI underwent successful primary angioplasty and systematic stenting with a Doppler guidewire. Patients with incomplete (<50%) STR 60 minutes after TIMI 3 flow was restored had flow velocity features suggestive of severe microcirculatory dysfunction, including a higher incidence of early systolic retrograde flow (41% versus 9%, P=0.007) and lower coronary flow velocity reserve (CVR, 1.3 versus 1.6, P<0.001). CVR improved immediately after stenting in patients with > or =50% STR but not in patients with <50% STR. There was a significant correlation between STR and poststent CVR. At 3 months, CVR was similar in patients with <50% and > or =50% STR. However, left ventriculography indicated lower global (42% versus 55%, P=0.001) and regional (16% versus 20%, P=0.03) left ventricular ejection fractions and 201Tl rest-redistribution scintigraphy indicated a larger infarct size (34% versus 16% 201Tl defect, P=0.007) in patients with <50% STR. CONCLUSIONS: After successful primary angioplasty with systematic stenting, <50% STR is a marker of severe albeit transient microcirculatory dysfunction in patients with AMI and is associated with more extensive myocardial damage. PMID- 12742981 TI - Safety and feasibility of catheter-based local intracoronary vascular endothelial growth factor gene transfer in the prevention of postangioplasty and in-stent restenosis and in the treatment of chronic myocardial ischemia: phase II results of the Kuopio Angiogenesis Trial (KAT). AB - BACKGROUND: Catheter-based intracoronary vascular endothelial growth factor (VEGF) gene transfer is a potential treatment for coronary heart disease. However, only limited data are available about local VEGF gene transfer given during angioplasty (PTCA) and stenting. METHODS AND RESULTS: Patients with coronary heart disease (n=103; Canadian Cardiovascular Society class II to III; mean age, 58+/-6 years) were recruited in this randomized, placebo-controlled, double-blind phase II study. PTCA was performed with standard methods, followed by gene transfer with a perfusion-infusion catheter. Ninety percent of the patients were given stents; 37 patients received VEGF adenovirus (VEGF-Adv, 2x10(10) pfu), 28 patients received VEGF plasmid liposome (VEGF-P/L; 2000 microg of DNA with 2000 microL of DOTMA:DOPE [1:1 wt/wt]), and 38 control patients received Ringer's lactate. Follow-up time was 6 months. Gene transfer to coronary arteries was feasible and well tolerated. The overall clinical restenosis rate was 6%. In quantitative coronary angiography analysis, the minimal lumen diameter and percent of diameter stenosis did not significantly differ between the study groups. However, myocardial perfusion showed a significant improvement in the VEGF-Adv-treated patients after the 6-month follow-up. Some inflammatory responses were transiently present in the VEGF-Adv group, but no increases were detected in the incidences of serious adverse events in any of the study groups. CONCLUSIONS: Gene transfer with VEGF-Adv or VEGF-P/L during PTCA and stenting shows that (1) intracoronary gene transfer can be performed safely (no major gene transfer-related adverse effects were detected), (2) no differences in clinical restenosis rate or minimal lumen diameter were present after the 6-month follow up, and (3) a significant increase was detected in myocardial perfusion in the VEGF-Adv-treated patients. PMID- 12742982 TI - Interleukin-7-mediated inflammation in unstable angina: possible role of chemokines and platelets. AB - BACKGROUND: Atherogenesis and plaque destabilization involve immune-mediated mechanisms, but the actual mediators have not been fully clarified. Interleukin (IL)-7 is a regulator of T-cell homeostasis but also may be involved in inflammation. We hypothesized that IL-7 could be involved in the inflammatory processes observed in atherosclerosis and acute coronary syndromes. METHODS AND RESULTS: To study the role of IL-7 in coronary artery disease, we analyzed IL-7 levels and the effect of this cytokine on inflammatory mediators in patients with stable and unstable angina and in healthy control subjects. Our major findings were (1) Plasma levels of IL-7 were significantly increased in patients with stable (n=30) and unstable angina (n=30) comparing healthy control subjects (n=20), particularly in those with unstable disease. (2) Increased release from activated platelets appeared to be a major contributor to the raised IL-7 levels in patients with angina. (3) IL-7 enhanced the expression of several inflammatory chemokines in peripheral blood mononuclear cells from both healthy control subjects and patients with angina, particularly in those with unstable disease. Similar effects were seen in monocytes but not in T cells. (4) MIP-1alpha further increased the release of IL-7 from platelets in a dose-dependent manner. (5) Aspirin reduced both the spontaneous and the SFLLRN-stimulated release of IL-7 from platelets, and when administered to healthy control subjects for 7 days (160 mg qd), it reduced plasma levels of IL-7. CONCLUSIONS: Our findings suggest a role for IL-7-driven inflammation in atherogenesis and the promotion of clinical instability in coronary artery disease involving interactions between platelets, monocytes, and chemokines. PMID- 12742983 TI - Cerebral microembolism is blocked by tirofiban, a selective nonpeptide platelet glycoprotein IIb/IIIa receptor antagonist. AB - BACKGROUND: Microembolic signals (MES) as detected by transcranial Doppler ultrasound define an individual stroke risk in patients with carotid artery disease. To study the composition of MES in vivo, we used the glycoprotein IIb/IIIa (GPIIb/IIIa) receptor antagonist tirofiban, a highly selective platelet aggregation inhibitor. METHODS AND RESULTS: Twenty-four patients with recent cerebral or retinal embolism of arterial origin and a MES rate >6 per hour on initial transcranial Doppler ultrasonography recording received the short-acting GPIIb/IIIa antagonist tirofiban. With tirofiban, the MES rate dropped from a median (range) of 38 (9 to 324) to zero in all patients. After cessation of infusion, the inhibitory effect of tirofiban was reversible, with a significant increase of MES (median 13.5; range, 0 to 35; n=16; P=0.001). Six patients received overlapping oral antiplatelet agents and remained MES-negative. CONCLUSIONS: Cerebral microembolism of arterial origin has the property of solid emboli, with platelet-fibrinogen units as predominant constituent parts. GPIIb/IIIa antagonists may have the potential to bridge the ischemic risk in patients with unstable carotid disease. PMID- 12742985 TI - Cardiopulmonary bypass to repair an atrial septal defect does not affect cognitive function in children. AB - BACKGROUND: Changes in neurocognitive function after cardiopulmonary bypass (CPB) are difficult to assess in children with congenital cardiovascular malformations in part because identification of a suitable control group of children is difficult. METHODS AND RESULTS: We evaluated neurocognitive function in subjects aged 3 to 17 years both before and after surgical repair of an atrial septal defect (ASD). Eighteen subjects underwent testing within a median of 3 days (range, 1 to 27) before surgery and 5.8 months (range, 5.5 to 9) after surgery. The Differential Ability Scales was administered to subjects in a single testing session. All subjects were considered to be healthy, and they had no activity limitations. The mean General Conceptual Ability (GCA) scores (analogous to IQ) for the group were in the normal range before and after ASD repair. No significant difference was detected between the GCA scores before and after surgery. The GCA score was 90.8+/-18.6 before surgery and 93.1+/-16.3 after surgery. CONCLUSIONS: Use of CPB was not associated with changes in cognitive function in our subjects who underwent repair of an ASD. PMID- 12742984 TI - Effects of intravenous nesiritide on human coronary vasomotor regulation and myocardial oxygen uptake. AB - BACKGROUND: Nesiritide, recombinant human B-type natriuretic peptide, has been shown to be efficacious in the treatment of decompensated heart failure. The effects of intravenous nesiritide on the human coronary vasculature have not been studied. METHODS AND RESULTS: Ten patients underwent right and left heart catheterization. Baseline coronary blood flow was determined using quantitative coronary angiography (QCA) and an intracoronary Doppler-tipped guidewire. Myocardial oxygen uptake was measured using a coronary sinus catheter. Patients then received an intravenous infusion of nesiritide (2 microg/kg bolus followed by 0.01 microg/kg per min infusion) for 30 minutes. Right atrial pressure decreased 52% (P=0.012), pulmonary artery mean pressure decreased 19% (P=0.03), pulmonary capillary wedge pressure decreased 46% (P=0.002), and mean arterial pressure decreased 11% (P=0.007). QCA demonstrated a 15% increase in coronary artery diameter from a baseline of 2.6+/-0.8 to 3.0+/-0.8 mm at 30 minutes (P=0.007). The coronary velocity measure of average peak velocity increased 14% from 20.8+/-6.4 at baseline to 23.8+/-7.2 cm/s at 5 minutes (P=0.015) and then returned to baseline for the remainder of the infusion. Coronary blood flow increased 35% (P=0.007), whereas coronary resistance decreased 23% at 15 and 30 minutes (P=0.036). Myocardial oxygen uptake decreased 8% during the nesiritide infusion (P=0.043). CONCLUSIONS: Nesiritide exerts coronary vasodilator effects on both the coronary conductance and resistance arteries. Despite a decrease in coronary perfusion pressure, coronary artery blood flow is increased, coronary resistance is decreased, and myocardial oxygen uptake is decreased. PMID- 12742986 TI - Ischemia modified albumin is a sensitive marker of myocardial ischemia after percutaneous coronary intervention. AB - BACKGROUND: Ischemia modified albumin (IMA; Ischemia Technologies, Inc) blood levels rise in patients who develop ischemia during percutaneous coronary intervention (PCI). It is not known whether IMA elevations correlate with increases in other markers of oxidative stress, ie, 8-iso prostaglandin F2-A (iP). METHODS AND RESULTS: We compared IMA versus iP plasma levels in 19 patients (mean age 62.8+/-11.9 years) undergoing PCI and 11 patients (mean age 64+/-13.6 years) undergoing diagnostic angiography (controls). In the PCI patients, blood samples for IMA and iP were taken from the guide catheter before PCI and after balloon inflations, and from the femoral sheath 30 minutes after PCI. IMA was measured by the albumin cobalt binding (ACB) test and plasma iP by enzyme immunoassay. During PCI, all 19 patients had chest pain and 18 had transient ischemic ST segment changes. IMA was elevated from baseline in 18 of the 19 patients after PCI. Median IMA levels were higher after PCI (101.4 U/mL, 95%CI 82 to 116) compared with baseline (72.8 U/mL, CI 55 to 93; P<0.0001). Levels remained elevated at 30 minutes (87.9 U/mL, CI 78 to 99; P<0.0001) and returned to baseline at 12 hours (70.3 U/mL, CI 65 to 87; P=0.65). iP levels were raised after PCI in 9 of the 19 patients. However, median iP levels were not significantly different immediately (P=0.6) or 30 minutes after PCI (P=0.1). In the control group, IMA and iP levels remained unchanged before and after angiography (P=0.2 and 0.16, respectively). CONCLUSIONS: IMA is a more consistent marker of ischemia than iP in patients who develop chest pain and ST segment changes during PCI. PMID- 12742987 TI - Prognostic role of brain natriuretic peptide in acute pulmonary embolism. AB - BACKGROUND: Rapid, noninvasive, and accurate prognostic assessment with an inexpensive cardiac biomarker is an appealing approach for patients with acute pulmonary embolism (PE). METHODS AND RESULTS: We measured at the time of admission the plasma level of plasma brain natriuretic peptide (BNP) to determine its utility in prognosticating the clinical course of 73 consecutive patients with acute PE. We used a prespecified BNP cut-off level (<90 pg/mL) for the prediction of the absence of a major adverse cardiovascular event, defined as any of the following: death, cardiopulmonary resuscitation, mechanical ventilation, or use of pressors, thrombolysis, catheter fragmentation, or surgical embolectomy. In the 20 (27%) patients with adverse events, median BNP (194.2, range 3.7 to 1201.1 pg/mL) was higher than in patients with a benign course (39.1, range 1.0 to 1560.0 pg/mL; P<0.001). However, 3 patients with adverse outcomes had low BNP levels on admission: 1 death, BNP 52 pg/mL; 1 patient with prolonged cardiopulmonary resuscitation, BNP 3.7 pg/mL; and 1 patient undergoing rescue thrombolysis, BNP 75 pg/mL. Sensitivity, specificity, and negative and positive predictive value of BNP levels <90 pg/mL for absence of adverse outcomes were 85% (64% to 95%), 75% (62% to 85%), 93% (95% CI 81% to 98%), and 57% (39% to 73%), respectively. The optimal BNP cut-off level, identified by receiver operating characteristic analysis, was <50 pg/mL. CONCLUSIONS: Low BNP levels do not guarantee an uncomplicated hospital course in patients with acute PE, using a "congestive heart failure" cut-off level of 90 pg/mL. A lower cut-off level of <50 pg/mL identifies 95% of patients with a benign clinical course. PMID- 12742988 TI - Inhibitory activity of clinical thiazolidinedione peroxisome proliferator activating receptor-gamma ligands toward internal mammary artery, radial artery, and saphenous vein smooth muscle cell proliferation. AB - BACKGROUND: The proliferation of vascular smooth muscle cells (VSMCs) is a known response to arterial injury that is an important part of the process of restenosis and atherosclerosis. People with diabetes have an increased risk of cardiovascular disease resulting from accelerated coronary atherosclerosis. The newest drugs for Type 2 diabetes are thiazolidinediones, which are insulin sensitizing peroxisome proliferator activating receptor-gamma (PPARgamma) ligands. We investigated the antiproliferative effects of troglitazone, rosiglitazone, and pioglitazone on VSMCs derived from the three vascular beds used for coronary artery by-pass grafting: the internal mammary and radial artery and saphenous veins. METHODS AND RESULTS: The three vessels yielded proliferating cells of slightly differing morphology. Inhibition of cell proliferation was assessed by cell counting and cell cycle studies by Western blotting for phosphorylated retinoblastoma protein. All three thiazolidinediones showed inhibitory potency toward cell proliferation with a potency troglitazone>rosiglitazone approximately pioglitazone, and this potency profile was maintained toward the growth factor and insulin-stimulated phosphorylation of the retinoblastoma protein, which controls cell cycle progression. CONCLUSIONS: The inhibitory potency of clinical thiazolidinediones toward different vascular sources is dependent on the individual thiazolidinedione and very little on the vascular source. PMID- 12742989 TI - Chymase inhibition prevents cardiac fibrosis and improves diastolic dysfunction in the progression of heart failure. AB - BACKGROUND: Angiotensin (Ang) II, which plays a crucial role in the cardiac remodeling process, is generated via angiotensin-converting enzyme (ACE); however, an alternative generation pathway, chymase, which is stored in the mast cells, also exists in the heart. Cardiac chymase is insensitive to ACE inhibitors (ACEIs), and heart chymase promotes interstitial fibrosis by affecting collagen metabolism via transforming growth factor-beta in vitro. Therefore, selective chymase blockade seems to be an important strategy in the prevention of cardiac remodeling METHODS AND RESULTS: We evaluated the effects of a specific chymase inhibitor, SUNC8257 (Chy I; 10 mg/kg twice a day; n=7), on changes in cardiac structures, Ang II levels, and gene expressions, which are characterized as molecular markers for fibrosis, in dogs with tachycardia induced heart failure (HF). In HF, the number of chymase enzyme-positive mast cells increased in the left ventricle (LV) compared with the normal group; however, Chy I significantly decreased the mast cell density and cardiac Ang II levels. Despite no significant differences in LV systolic function compared with the vehicle group, Chy I decreased LV end-diastolic pressure and shortened the prolongation of tau. Chy I suppressed collagen-type I and III and transforming growth factor-beta mRNA levels and decreased fibrosis in the LV compared with the vehicle. CONCLUSIONS: The chymase pathway may be critical for cardiac diastolic dysfunction accompanied with fibrosis. Chronic chymase inhibition may therefore become an important strategy in the prevention of cardiac remodeling in HF. PMID- 12742990 TI - Intramural delivery of recombinant apolipoprotein A-IMilano/phospholipid complex (ETC-216) inhibits in-stent stenosis in porcine coronary arteries. AB - BACKGROUND: We have previously demonstrated vasculoprotective effects after repeated intravenous administration of recombinant apolipoprotein A-IMilano (apoA Im)/phospholipid complex. In this study, we sought to determine the effects of local recombinant apoA-Im/1-palmitoyl,2-oleoyl phosphatidylcholine complex (ETC 216) delivered intramurally via the Infiltrator catheter on luminal narrowing in a porcine coronary artery stent overstretch injury model. METHODS AND RESULTS: In twelve domestic swine ( approximately 25 kg), two arteries each were infiltrated with 0.4 mL ETC-216 (14 mg/mL) or vehicle control immediately before deployment of GFX stents (stent:artery ratio=1.3:1). Animals were euthanized at day 28, and evaluation by QCA revealed a significant improvement in mean lumen loss index with ETC-216 treatment (21+/-22% versus 43+/-13% lumen loss; P=0.01). Histomorphometric analysis showed that ETC-216 treatment significantly reduced the intimal area (6.7+/-1.5 versus 5.2+/-1.4 mm2, -22%; P=0.02) and the stenosis index (0.76+/-0.15 versus 0.59+/-0.15; P=0.01), and increased the lumen area (2.1+/-1.4 versus 3.7+/-1.8 mm2, +76%; P=0.02). Regression analysis showed significant differences in lumen area (P=0.004), neointimal area (P=0.003), stenosis index (P=0.001), and neointimal thickness (P=0.003) adjusted for injury score in favor of ETC-216. CONCLUSIONS: A single intramural administration of ETC 216 significantly inhibited injury-induced luminal narrowing in the porcine stent overstretch model through reduction of intimal hyperplasia. These data show that local intracoronary delivery of ETC-216 may be useful to prevent restenosis after coronary stenting. PMID- 12742991 TI - Elevated plasma levels of the atherogenic mediator soluble CD40 ligand in diabetic patients: a novel target of thiazolidinediones. AB - BACKGROUND: Considerable evidence implicates the proinflammatory cytokine CD40 ligand (CD40L) in atherosclerosis and accumulating data link type 1 and 2 diabetes, conditions associated with accelerated atherosclerosis, to inflammation. This study therefore evaluated the hypothesis that diabetic patients have elevated plasma levels of soluble CD40L (sCD40L) and that treatment with the insulin-sensitizing thiazolidinediones lowers this index of inflammation. METHODS AND RESULTS: Subjects with type 1 (n=49) or type 2 diabetes (n=48) had higher (P<0.001) sCD40L plasma levels (6.56+/-3.27 and 6.67+/-2.90 ng/mL, respectively) compared with age-matched control groups (1.40+/-2.21 and 1.32+/-2.68 ng/mL, respectively). Multiple regression analysis demonstrated a significant (P<0.001) association between plasma sCD40L and type 1 as well as type 2 diabetes, independent of total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triglycerides, blood pressure, body mass index, gender, C-reactive protein, and soluble intracellular adhesion molecule-1. Furthermore, in a pilot study, administration of troglitazone (12 weeks, 600 mg/day), but not placebo, to type 2 diabetics (n=68) significantly (P<0.001) diminished sCD40L plasma levels by 29%. The thiazolidinedione lowered plasma sCD40L in type 2 diabetic patients with long-standing disease (>3 years) with or without macrovascular complications (-34% and -29%, respectively) as well as in type 2 diabetic patients with more recent (<3 years) onset of the disease ( 27%; all P<0.05). CONCLUSIONS: This study provides new evidence that individuals with type 1 or 2 diabetes have a proinflammatory state as indicated by elevated levels of plasma sCD40L. Troglitazone treatment of type 2 diabetic patients diminishes sCD40L levels, suggesting a novel antiinflammatory mechanism for limiting diabetes-associated arterial disease. PMID- 12742993 TI - Genome-wide array analysis of normal and malformed human hearts. AB - BACKGROUND: We present the first genome-wide cDNA array analysis of human congenitally malformed hearts and attempted to partially elucidate these complex phenotypes. Most congenital heart defects, which account for the largest number of birth defects in humans, represent complex genetic disorders. As a consequence of the malformation, abnormal hemodynamic features occur and cause an adaptation process of the heart. METHODS AND RESULTS: The statistical analysis of our data suggests distinct gene expression profiles associated with tetralogy of Fallot, ventricular septal defect, and right ventricular hypertrophy. Applying correspondence analysis, we could associate specific gene functions to specific phenotypes. Furthermore, our study design allows the suggestion that alterations associated with primary genetic abnormalities can be distinguished from those associated with the adaptive response of the heart to the malformation (right ventricular pressure overload hypertrophy). We provide evidence for the molecular transition of the hypertrophic right ventricle to normal left ventricular characteristics. Furthermore, we present data on chamber-specific gene expression. CONCLUSIONS: Our findings propose that array analysis of malformed human hearts opens a new window to understand the complex genetic network of cardiac development and adaptation. For detailed access, see the online-only Data Supplement. PMID- 12742992 TI - Differentiation of human embryonic stem cells to cardiomyocytes: role of coculture with visceral endoderm-like cells. AB - BACKGROUND: Cardiomyocytes derived from human embryonic stem (hES) cells could be useful in restoring heart function after myocardial infarction or in heart failure. Here, we induced cardiomyocyte differentiation of hES cells by a novel method and compared their electrophysiological properties and coupling with those of primary human fetal cardiomyocytes. METHODS AND RESULTS: hES cells were cocultured with visceral-endoderm (VE)-like cells from the mouse. This initiated differentiation to beating muscle. Sarcomeric marker proteins, chronotropic responses, and ion channel expression and function were typical of cardiomyocytes. Electrophysiology demonstrated that most cells resembled human fetal ventricular cells. Real-time intracellular calcium measurements, Lucifer yellow injection, and connexin 43 expression demonstrated that fetal and hES derived cardiomyocytes are coupled by gap junctions in culture. Inhibition of electrical responses by verapamil demonstrated the presence of functional alpha1c calcium ion channels. CONCLUSIONS: This is the first demonstration of induction of cardiomyocyte differentiation in hES cells that do not undergo spontaneous cardiogenesis. It provides a model for the study of human cardiomyocytes in culture and could be a step forward in the development of cardiomyocyte transplantation therapies. PMID- 12742994 TI - Elevated production of interleukin-6 is associated with a lower incidence of disease-related ischemic events in patients with giant-cell arteritis: angiogenic activity of interleukin-6 as a potential protective mechanism. AB - BACKGROUND: Patients with giant-cell arteritis (GCA) who develop a strong acute phase response are at low risk of disease-related ischemic events. METHODS AND RESULTS: To assess the potential protective role of proinflammatory cytokines in the development of ischemic events in GCA, we measured tissue expression (66 individuals) and/or circulating levels (80 individuals) of interleukin (IL) 1beta, tumor necrosis factor-alpha (TNF-alpha), and IL-6 in patients with biopsy proven GCA. Tissue expression was determined by quantitative real-time polymerase chain reaction and immunohistochemistry. Circulating cytokines were determined by enzyme-linked immunoassay. We found that patients with disease-related ischemic events had lower IL-6 mRNA levels (5.9+/-2.1 versus 27.6+/-7.8 relative units, P=0.013), lower IL-6 immunohistochemical expression scores (1.5+/-0.9 versus 2.7+/-1, P=0.001), and lower circulating levels of IL-6 (13.6+/-2.1 versus 24+/ 2.4 pg/mL, P=0.002) than patients without ischemic complications. No significant differences were found for either IL-1beta or TNF-alpha. We subsequently investigated direct effects of IL-6 on vessel wall components. We found that IL-6 stimulates endothelial cell proliferation and differentiation into capillary-like structures and induces full angiogenic activity in both ex vivo (aortic ring) and in vivo (chick chorioallantoic membrane) assays. CONCLUSIONS: GCA patients with ischemic complications have lower tissue expression and circulating levels of IL 6 than patients with no ischemic events. IL-6 has relevant direct effects on vascular wall components that might be protective: IL-6 activates a functional program related to angiogenesis that may compensate for ischemia in patients with GCA. PMID- 12742995 TI - Critical role of L-arginine in endothelial cell survival during oxidative stress. AB - BACKGROUND: Oxidative damage of vascular endothelium represents an important initiation step in the development of atherosclerosis. Recently, we reported about protection of inducible nitric oxide synthase (iNOS)-derived high-output NO in endothelial cells. Because iNOS activity critically depends on the availability of its substrate l-arginine, the present study aims at elucidating iNOS-mediated effects on H2O2-induced apoptosis of cytokine-activated rat aortic endothelial cells (AECs) subject to medium l-arginine concentrations. METHODS AND RESULTS: In cytokine-activated AECs, iNOS activity was found to be half-maximal at 60 micromol/L arginine, which represents the medium serum level in rats but also in humans. Maximal activity is seen at and above 200 micromol/L arginine. Activated cells grown in the absence of arginine with minimal iNOS activity are highly sensitive toward H2O2-induced apoptosis, and increases in medium arginine concentrations result in increased cell survival. Moreover, competition experiments show that iNOS activity is completely dependent on cationic amino acid transporter-mediated arginine uptake. We also find that the arginine dependent protection includes inhibition of endothelial lipid peroxidation and increases in the expression of vasoprotective stress response genes. CONCLUSIONS: Our data demonstrate that arginine concentrations corresponding to physiological serum levels do not allow for optimal endothelial iNOS activity. Thus, decreases in systemic arginine concentrations, or locally within atherosclerotic plaques, will impair the endothelial iNOS-mediated stress response and will significantly increase the risk of endothelial dysfunction. PMID- 12742996 TI - Third-generation beta-blockers stimulate nitric oxide release from endothelial cells through ATP efflux: a novel mechanism for antihypertensive action. AB - BACKGROUND: Nebivolol and carvedilol are third-generation beta-adrenoreceptor antagonists, which unlike classic beta-blockers, have additional endothelium dependent vasodilating properties specifically related to microcirculation by a molecular mechanism that still remains unclear. We hypothesized that nebivolol and carvedilol stimulate NO release from microvascular endothelial cells by extracellular ATP, which is a well-established potent autocrine and paracrine signaling factor modulating a variety of cellular functions through the activation of P2-purinoceptors. METHODS AND RESULTS: Contraction and relaxation of renal glomerular vasculature were measured by determination of intracapillary volume with [3H]-inulin. Biologically active NO was measured with highly sensitive porphyrinic NO microsensors in a single glomerular endothelial cell (GEC). Extracellular ATP was measured by a luciferin-luciferase assay. Enzymatic degradation of extracellular ATP by apyrase and blockade of P2Y-purinoceptors by suramin or reactive blue 2 inhibited both beta-blocker-induced glomerular vasorelaxations and beta-blocker-stimulated NO release from GECs. Both beta blocker-induced vasorelaxations were in the micromolar concentration range identical to that required for the beta-blocker stimulation of ATP and NO release from GECs. The maximum of NO release for nebivolol and carvedilol was very similar (188+/-14 and 226+/-17, respectively). Blockade of ATP release by a mechanosensitive ion channel blocker, Gd3+, inhibited the beta-blocker-dependent release of ATP and NO from GECs. CONCLUSIONS: These results demonstrate for the first time that nebivolol and carvedilol induce relaxation of renal glomerular microvasculature through ATP efflux with consequent stimulation of P2Y purinoceptor-mediated NO release from GECs. PMID- 12742997 TI - Long-term stable expression of human apolipoprotein A-I mediated by helper dependent adenovirus gene transfer inhibits atherosclerosis progression and remodels atherosclerotic plaques in a mouse model of familial hypercholesterolemia. AB - BACKGROUND: Epidemiologic studies and transgenic mouse experiments indicate that high plasma HDL and apolipoprotein (apo) A-I protect against atherosclerosis. We used helper-dependent adenovirus (HD-Ad) gene transfer to examine the effect of long-term hepatic apoA-I expression on atherosclerotic lesion progression and remodeling in a mouse model of familial hypercholesterolemia. METHODS AND RESULTS: We treated LDL receptor-deficient (LDLR-/-) mice maintained on a high cholesterol diet for 6 weeks with either a HD-Ad containing human apoA-I gene (HD Ad-AI) or saline (control). HD-Ad-AI treatment did not affect plasma liver enzymes but induced the appearance of plasma human apoA-I at or above human levels for the duration of the study. Substantial amounts of human apoA-I existed in lipid-free plasma. Compared with controls, HDLs from treated mice were larger and had a greater inhibitory effect on tumor necrosis factor-alpha-induced vascular cellular adhesion molecule-1 expression in cultured endothelial cells. Twenty-four weeks after injection, aortic atherosclerotic lesion area in saline treated mice progressed approximately 700%; the rate of progression was reduced by >50% by HD-Ad-AI treatment. The lesions in HD-Ad-AI-treated mice contained human apoA-I that colocalized mainly with macrophages; they also contained less lipid, fewer macrophages, and less vascular cellular adhesion molecule-1 immunostaining but more smooth muscle cells (alpha-actin staining) and collagen. CONCLUSIONS: HD-Ad-AI treatment of LDLR-/- mice leads to long-term overexpression of apoA-I, retards atherosclerosis progression, and remodels the lesions to a more stable-appearing phenotype. HD-Ad-mediated transfer of apoA-I may be a useful clinical approach for protecting against atherosclerosis progression and stabilizing atherosclerotic lesions associated with dyslipidemia in human patients. PMID- 12742998 TI - Augmentation of wall shear stress inhibits neointimal hyperplasia after stent implantation: inhibition through reduction of inflammation? AB - BACKGROUND: Low wall shear stress (WSS) increases neointimal hyperplasia (NH) in vein grafts and stents. We studied the causal relationship between WSS and NH formation in stents by locally increasing WSS with a flow divider (Anti Restenotic Diffuser, Endoart SA) placed in the center of the stent. METHODS AND RESULTS: In 9 rabbits fed a high-cholesterol diet for 2 months to induce endothelial dysfunction, 18 stents were implanted in the right and left external iliac arteries (1 stent per vessel). Lumen diameters were measured by quantitative angiography before and after implantation and at 4-week follow-up, at which time, macrophage accumulation and interruption of the internal elastic lamina was determined. Cross sections of stent segments within the ARED (S+ARED), outside the ARED (S[minus]ARED), and in corresponding segments of the contralateral control stent (SCTRL) were analyzed. Changes in WSS induced by the ARED placement were derived by computational fluid dynamics. Computational fluid dynamics analysis demonstrated that WSS increased from 0.38 to 0.82 N/m2 in the S+ARED immediately after ARED placement. This augmentation of shear stress was accompanied by (1) lower mean late luminal loss by quantitative angiography ([minus]0.23+/-0.22 versus [minus]0.58+/-0.30 mm, P=0.02), (2) reduction in NH (1.48+/-0.58, 2.46+/-1.25, and 2.36+/-1.13 mm2, P<0.01, respectively, for S+ARED, S[minus]ARED, and SCTRL), and (3) a reduced inflammation score and a reduced injury score. Increments in shear stress did not change the relationship between injury score and NH or between inflammation score and NH. CONCLUSIONS: The newly developed ARED flow divider significantly increases WSS, and this local increment in WSS is accompanied by a local reduction in NH and a local reduction in inflammation and injury. The present study is therefore the first to provide direct evidence for an important modulating role of shear stress in in-stent neointimal hyperplasia. PMID- 12742999 TI - Variants of toll-like receptor 4 modify the efficacy of statin therapy and the risk of cardiovascular events. AB - BACKGROUND: Atherosclerosis is increasingly considered to be a chronic inflammatory process. We examined whether genetic variants of the toll-like receptor 4 (TLR4), which are correlated with impaired innate immunity and with progression of carotid atherosclerosis, are also associated with coronary atherosclerosis and predict the risk of cardiovascular events. METHODS AND RESULTS: Two polymorphisms of the TLR4 gene (Asp299Gly and Thr399Ile) were determined in 655 men with angiographically documented coronary atherosclerosis. All patients participated in a prospective cholesterol-lowering trial evaluating the effect on coronary artery disease and were randomly assigned to either pravastatin or placebo for 2 years. There were no significant differences between genetically defined subgroups with respect to baseline risk factors, treatment, or in-trial changes of lipid, lipoprotein, or angiographic measurements. Genotype was not associated with progression of atherosclerosis. In the pravastatin group, 299Gly carriers had a lower risk of cardiovascular events during follow-up than noncarriers (2.0% versus 11.5%, P=0.045). Among noncarriers, pravastatin reduced the risk of cardiovascular events from 18.1% to 11.5% (P=0.03), whereas among 299Gly carriers this risk was strikingly reduced from 29.6% to 2.0% (P=0.0002, P=0.025 for interaction). CONCLUSIONS: Among symptomatic men with documented coronary artery disease, the TLR4 Asp299Gly polymorphism was associated with the risk of cardiovascular events. This variant also modified the efficacy of pravastatin in preventing cardiovascular events, such that carriers of the variant allele had significantly more benefit from pravastatin treatment. PMID- 12743000 TI - Longitudinal myocardial function assessed by tissue velocity, strain, and strain rate tissue Doppler echocardiography in patients with AL (primary) cardiac amyloidosis. AB - BACKGROUND: AL amyloidosis with heart failure is associated with decreased longitudinal myocardial contraction measured by pulsed tissue Doppler imaging. We sought to clarify whether new modalities of myocardial strain Doppler (change in length per unit length) or strain rate (the temporal derivative of strain) were more sensitive than tissue Doppler and could detect early regional myocardial dysfunction before the onset of congestive heart failure (CHF) in patients with AL (primary) amyloidosis. METHODS AND RESULTS: Ninety-seven biopsy-proven patients with AL amyloidosis were divided into 3 groups. Group 1 patients had no cardiac involvement (n=36), group 2 had heart involvement but no CHF (n=32), and group 3 had heart involvement and CHF (n=29). All patients underwent tissue velocity (TV) imaging, strain, and strain rate imaging (SR) at the basal, mid, and apical ventricle in 2 apical views. With the use of TV, differences in systolic function were only apparent between group 3 (basal mean value, 3.0+/-1.1 cm/s) and groups 1 and 2 (5.0+/-1.3 and 4.6+/-1.2 cm/s, respectively). In contrast, basal peak systolic SR (l/s) showed significant differences among all 3 groups (-2.0+/-0.4, -1.55+/-0.6, and -0.76+/-0.3 for groups 1 to 3, respectively. P<0.01). Basal strain also demonstrated statistically significant differences among the groups (-19+/-4%, -15+/-4.5%, and -8.0+/-5%; P<0.01). CONCLUSIONS: Cardiac amyloidosis is characterized by an early impairment in systolic function at a time when fractional shortening remains normal. This abnormality precedes the onset of CHF and can be detected by strain and SR but is not apparent by TV imaging. PMID- 12743001 TI - Beta-blockers restore calcium release channel function and improve cardiac muscle performance in human heart failure. AB - BACKGROUND: Chronic beta-adrenergic receptor (beta-AR) blockade improves cardiac contractility and prolongs survival in patients with heart failure; however, the mechanisms underlying these favorable responses are poorly understood. Stress induced activation of the sympathetic nervous system results in protein kinase A (PKA)-mediated phosphorylation of the calcium (Ca2+) release channel/cardiac ryanodine receptor (RyR2), required for cardiac excitation-contraction (EC) coupling, activating the RyR2 channel, and increasing cardiac contractility. The hyperadrenergic state of heart failure results in leaky RyR2 channels attributable to PKA hyperphosphorylation and depletion of the stabilizing FK506 binding protein, FKBP12.6. We tested the hypothesis that improved cardiac muscle function attributable to beta-AR blockade is associated with restoration of normal RyR2 channel function in patients with heart failure. METHODS AND RESULTS: We assessed the effects of beta-AR blockade on left ventricular volume using isolated perfused hearts and beta-agonist responsiveness using muscle strips from patients undergoing transplantation. Twenty-four human hearts were examined, 10 from patients with heart failure treated with beta-AR blockers (carvedilol, metoprolol, or atenolol), 9 from patients with heart failure without beta-AR blocker treatment, and 5 normal hearts. RyR2 PKA phosphorylation was determined by back-phosphorylation, FKBP12.6 in the RyR2 macromolecular complex was determined by coimmunoprecipitation, and channel function was assayed using planar lipid bilayers. beta-AR blockers reduced left ventricular volume (reverse remodeling) and restored beta-agonist response in cardiac muscle from patients with heart failure. Improved cardiac muscle function was associated with restoration of normal FKBP12.6 levels in the RyR2 macromolecular complex and RyR2 channel function. CONCLUSIONS: Improved cardiac muscle function during beta-AR blockade is associated with improved cardiac Ca2+ release channel function in patients with heart failure. PMID- 12743002 TI - Obstructive sleep apnea and the recurrence of atrial fibrillation. AB - BACKGROUND: We tested the hypothesis that patients with untreated obstructive sleep apnea (OSA) would be at increased risk for recurrence of atrial fibrillation (AF) after cardioversion. METHODS AND RESULTS: We prospectively obtained data on history, echocardiogram, ECG, body mass index, hypertension, diabetes, NYHA functional class, ejection fraction, left atrial appendage velocity, and medications in patients with AF/atrial flutter referred for DC cardioversion. Forty-three individuals were identified as having OSA on the basis of a previous sleep study. Data regarding the use of continuous positive airway pressure (CPAP) and recurrence of AF were obtained for 39 of these patients. Follow-up data were also obtained in 79 randomly selected postcardioversion patients (controls) who did not have any previous sleep study. Twenty-seven of the 39 OSA patients either were not receiving any CPAP therapy (n=25) or were using CPAP inappropriately (n=2). Recurrence of AF at 12 months in these 27 patients was 82%, higher than the 42% recurrence in the treated OSA group (n=12, P=0.013) and the 53% recurrence (n=79, P=0.009) in the 79 control patients. Of the 25 OSA patients who had not been treated at all, the nocturnal fall in oxygen saturation was greater (P=0.034) in those who had recurrence of AF (n=20) than in those without recurrence (n=5). CONCLUSIONS: Patients with untreated OSA have a higher recurrence of AF after cardioversion than patients without a polysomnographic diagnosis of sleep apnea. Appropriate treatment with CPAP in OSA patients is associated with lower recurrence of AF. PMID- 12743003 TI - Synergistic effect of persistent Chlamydia pneumoniae infection, autoimmunity, and inflammation on coronary risk. AB - BACKGROUND: Given the role of chronic infections, autoimmunity, and inflammation in atherosclerosis, we studied the joint effect of chronic Chlamydia pneumoniae infection, persistently elevated human heat-shock protein 60 (hHsp60) antibodies, and C-reactive protein (CRP) on coronary risk. METHODS AND RESULTS: The participants for this prospective nested case-control study were obtained from the Helsinki Heart Study, during which 241 nonfatal myocardial infarctions or coronary deaths occurred among 4081 dyslipidemic middle-aged men. Serum samples taken at baseline and 3 to 6 months before the coronary events that occurred during the 8.5-year period were analyzed for antibodies to C pneumoniae and hHsp60 and the CRP concentration. Compared with persistently low levels, the risk of coronary events was 2-fold for persistently elevated immunocomplex (IC)-bound and/or serum IgA antibodies to C pneumoniae (OR, 1.96; 95% CI, 1.14 to 3.36) and also for serum IgA antibodies to hHsp60 (OR, 2.11; 95% CI, 1.08 to 4.13). The risks associated with elevated antibodies were much higher when CRP was also elevated. Compared with low or transiently elevated levels, the risk of coronary events, with adjustment for age and smoking, was 4.5-fold for persistently elevated CRP and C pneumoniae IC/IgA antibodies together (OR, 4.47; 95% CI, 1.84 to 10.83) and was similar for CRP and hHsp60 IgA antibodies together (OR, 4.36; 95% CI, 1.53 to 12.39). CONCLUSIONS: Persistently but not transiently elevated C pneumoniae IC/IgA and hHsp60 IgA antibodies, especially when present together with an elevated CRP level, predicted coronary events. PMID- 12743004 TI - Effects of continuous enhanced vagal tone on dual atrioventricular node and accessory pathways. AB - BACKGROUND: The aim of this study was to test the electrophysiological effects of continuous enhanced vagal tone on dual atrioventricular (AV) nodal and accessory pathways. METHODS AND RESULTS: This study included 10 patients with typical, slow fast AV nodal reentrant tachycardia (AVNRT) and 10 patients with AV reciprocating tachycardia. Electrophysiological data were measured before and during continuous vagal enhancement by using phenylephrine infusion (0.6 to 1.5 microg/kg per min). For patients with AVNRT, during phenylephrine infusion, 1:1 conduction times over the anterograde fast and slow and retrograde fast pathways were prolonged (453+/ 64 to 662+/-120 ms, P<0.001; 379+/-53 to 443+/-95 ms, P<0.05; 405+/-112 to 442+/ 118 ms, P<0.05). The effective refractory period and functional refractory period of the anterograde fast pathway were prolonged with phenylephrine (394+/-73 to 544+/-128 ms, P<0.001; 454+/-60 to 596+/-118 ms, P<0.001). In contrast, the effective refractory period and functional refractory period of the anterograde slow and retrograde fast were not significantly changed. No significant change was observed in the conduction or refractoriness of the accessory pathways in patients with AV reciprocating tachycardia nor in atrial or ventricular refractoriness. CONCLUSIONS: Enhanced vagal tone produces disparate effects on the refractoriness of the slow and fast AV nodal conduction pathways, with the anterograde fast pathway being the most sensitive. These changes are conducive to induction of AVNRT with a premature atrial complex and may explain in part the relatively common occurrence of AVNRT during sleep or other periods of presumed increased parasympathetic tone. PMID- 12743005 TI - Electron-beam tomography coronary artery calcium and cardiac events: a 37-month follow-up of 5635 initially asymptomatic low- to intermediate-risk adults. AB - BACKGROUND: Conventional coronary artery disease (CAD) risk factors fail to explain nearly 50% of CAD events. This study examines the association between electron-beam tomography (EBT) coronary artery calcium (CAC) and cardiac events in initially asymptomatic low- to intermediate-risk individuals, with adjustment for the presence of hypercholesterolemia, hypertension, diabetes, and a history of cigarette smoking. METHODS AND RESULTS: The study was performed in 8855 initially asymptomatic adults 30 to 76 years old (26% women) who self-referred for EBT CAC screening. Conventional CAD risk factors were elicited by use of a questionnaire. After 37+/-12 months, information on the occurrence of cardiac events was collected and confirmed by use of medical records and death certificates. In men, events (n=192) were associated with the presence of CAC (RR=10.5, P<0.001), diabetes (RR=1.98, P=0.008), and smoking (RR=1.4, P=0.025), whereas in women, events (n=32) were linked to the presence of CAC (RR=2.6, P=0.037) and not risk factors. The presence of CAC provided incremental prognostic information in addition to age and other risk factors. CONCLUSIONS: The association between EBT CAC and cardiac events observed in this study of initially asymptomatic, middle-aged, low to intermediate-risk individuals presenting for screening suggests that in this group, knowledge of the presence of EBT CAC provides incremental information in addition to that defined by conventional CAD risk assessment. PMID- 12743006 TI - Sudden death in noncoronary heart disease is associated with delayed paced ventricular activation. AB - BACKGROUND: Slowed or delayed myocardial activation and dispersed refractoriness predispose to reentrant excitation that may lead to ventricular fibrillation (VF). Increased ventricular electrogram duration (DeltaED) in response to extrastimuli and increased S1S2 coupling intervals at which electrogram duration starts to increase (S1S2delay) are seen both in hypertrophic cardiomyopathy (HCM) in those at risk of VF and in patients with idiopathic VF (IVF). METHODS AND RESULTS: DeltaED and S1S2delay have been measured using paced electrogram fractionation analysis in 266 patients with noncoronary heart disease. Of these, one group of 61 patients had a history of VF and included 21 HCM, 17 IVF, 13 long QT syndrome (LQTS), 5 dilated cardiomyopathy (DCM), and 5 others. These were compared with 205 patients with similar diseases with no VF history (non-VF group) and a control group (n=12) without heart disease. Results from HCM VF patients (DeltaED, 19+/-3.3 ms; S1S2delay, 350+/-9.7 ms) differed sharply from observations in HCM non-VF patients (DeltaED, 7.3+/-1.35 ms; S1S2delay, 312+/-6.7 ms; P<0.001). DCM VF patients had longer delays (DeltaED, 14.3+/-5.9; S1S2delay, 344+/-11.2) than DCM non-VF patients (DeltaED, 5.8+/-1.87 ms; S1S2delay, 311+/ 5.7 ms; P<0.001), with major differences also seen comparing LQTS VF (DeltaED, 12.4+/-5.3 ms; S1S2delay, 343+/-13.8 ms) and LQTS non-VF patients (DeltaED, 11.0+/-2.7 ms; S1S2delay, 320+/-5.4 ms; P<0.001). IVF patients had both severely abnormal and normal areas of myocardium. CONCLUSIONS: Slowed or delayed myocardial activation is a common feature in patients with noncoronary heart disease with a history of VF, and its assessment may allow the prospective prediction of VF risk in these patients. PMID- 12743007 TI - An underrecognized subepicardial reentrant ventricular tachycardia attributable to left ventricular aneurysm in patients with normal coronary arteriograms. AB - BACKGROUND: In patients with apparently normal hearts, ventricular tachycardia (VT) may only involve the subepicardial myocardium. METHODS AND RESULTS: Four patients with exercise-induced fast VT with right bundle branch block morphology were investigated. ECG showed a small q wave in leads II, III, and aVF during sinus rhythm (SR) in all 4 patients. Left ventricular angiography showed small inferolateral aneurysms in all patients. Coronary arteriograms were normal in all 4 patients. Six unstable VTs (cycle length, 200 to 305 ms) and 1 stable VT (cycle length 370 ms) were reproducibly induced in the 4 patients. During SR, endocardial mapping was normal in all 4 patients, and epicardial mapping showed fragmented and late potentials in the left inferolateral wall anatomically consistent with the left ventricle aneurysm. During tachycardia, epicardial mapping showed a macroreentrant VT with focal endocardial activation in the patient with stable VT, whereas in 2 patients with unstable VT, a diastolic potential was only recorded and coincided with the late potential in the same area. Epicardial ablation was performed in 3 patients and successfully abolished those VTs. No VT recurred in 2 patients during follow-up of 2 and 9 months. Clinical VT recurred 6 months after the ablation and was successfully ablated in a repeated epicardial ablation in 1 patient. In the remaining patient without epicardial ablation, an implantable cardiac defibrillator was implanted. There were multiple shocks during a follow-up of 31 months. CONCLUSIONS: In patients with normal coronary arteriograms and left ventricle aneurysm, exercise-induced VT with right bundle branch block morphology may have a subepicardial arrhythmogenic substrate, which may be amenable to epicardial ablation. PMID- 12743008 TI - NO- activates soluble guanylate cyclase and Kv channels to vasodilate resistance arteries. AB - Nitric oxide (NO) plays an important role in the control of vascular tone. Traditionally, its vasorelaxant activity has been attributed to the free radical form of NO (NO*), yet the reduced form of NO (NO-) is also produced endogenously and is a potent vasodilator of large conduit arteries. The effects of NO- in the resistance vasculature remain unknown. This study examines the activity of NO- in rat small isolated mesenteric resistance-like arteries and characterizes its mechanism(s) of action. With the use of standard myographic techniques, the vasorelaxant properties of NO* (NO gas solution), NO- (Angeli's salt), and the NO donor sodium nitroprusside were compared. Relaxation responses to Angeli's salt (pEC50=7.51+/-0.13, Rmax=95.5+/-1.5%) were unchanged in the presence of carboxy PTIO (NO* scavenger) but those to NO* and sodium nitroprusside were inhibited. l Cysteine (NO- scavenger) decreased the sensitivity to Angeli's salt (P<0.01) and sodium nitroprusside (P<0.01) but not to NO*. The soluble guanylate cyclase inhibitor ODQ (3 and 10 micromol/L) concentration-dependently inhibited relaxation responses to Angeli's salt (41.0+/-6.0% versus control 93.4+/-1.9% at 10 micromol/L). The voltage-dependent K+ channel inhibitor 4-aminopyridine (1 mmol/L) caused a 9-fold (P<0.01) decrease in sensitivity to Angeli's salt, whereas glibenclamide, iberiotoxin, charybdotoxin, and apamin were without effect. In combination, ODQ and 4-aminopyridine abolished the response to Angeli's salt. In conclusion, NO- functions as a potent vasodilator of resistance arteries, mediating its response independently of NO* and through the activation of soluble guanylate cyclase and voltage-dependent K+ channels. NO- donors may represent a novel class of nitrovasodilator relevant for the treatment of cardiovascular disorders such as angina. PMID- 12743009 TI - Angiotensinogen single nucleotide polymorphisms, elevated blood pressure, and risk of cardiovascular disease. AB - In this study of 10 690 individuals, associations with elevated blood pressure, ischemic heart disease, and ischemic cerebrovascular disease were determined for two noncoding [A(-20)C, G(-6)A] and two coding (T174M, M235T) single nucleotide polymorphisms, analyzed alone and in combination (haplotypes). Participants from the general population with (n=4950) and without (n=4234) elevated blood pressure were compared (study 1), as were participants from the general population without ischemic heart disease and ischemic cerebrovascular disease (n=7965) and cases with either ischemic heart disease (n=1850, study 2) or ischemic cerebrovascular disease (n=848, study 3). Finally, 22-year follow-up of 9184 individuals from the general population examined risk of ischemic heart disease (study 4) and ischemic cerebrovascular disease (study 5). Individuals with -6AA, 174TT, or 235TT had plasma angiotensinogen levels increased by 80 ng/mL (P=0.01 and 0.05 for women and men) compared with individuals with -6GG, 174TT, or 235 MM. In women, this difference was associated with an odds ratio of elevated blood pressure of 1.25 (1.03 to 1.51), which increased to 1.63 (1.05 to 2.51) in postmenopausal women receiving hormone replacement therapy. The promoter single nucleotide polymorphisms alone or as haplotypes did not predict the continuous variables of systolic, diastolic, or pulse pressure in cross section or the risk of ischemic heart disease or ischemic cerebrovascular disease in either gender in case control or prospective studies. Individuals with -6AA, 174TT, or 235TT in the angiotensinogen gene have increased plasma angiotensinogen levels and moderately increased risk of elevated blood pressure (women only) but unaltered blood pressure examined as a continuous variable and unaltered risk of ischemic heart disease and ischemic cerebrovascular disease. PMID- 12743011 TI - Effects of endothelin-1 and endothelin-1 receptor blockade on cardiac output, aortic pressure, and pulse wave velocity in humans. AB - Endothelin-1 (ET-1) is a potent vasoconstrictor. Its effect on arterial wave reflections and central pressure augmentation is unknown. We studied whether ET 1, in plasma concentrations present in disease, increases pulse wave velocity (PWV) and augmentation index (AIx) and therefore compromises cardiac output, and whether the ET-1 receptor blocker VML-588 (previously AXV-034343 and Ro 61-1790) prevents such effects. Nine healthy men received a 2-hour infusion with ET-1 (2.5 ng x kg(-1) x min(-1)) superimposed on vehicle or VML-588 (0.05, 0.20, or 0.40 mg x kg(-1) x h(-1)) (randomized order). Arterial tonometry and pulse wave contour analysis were used to assess aortic PWV and central aortic pressures and impedance cardiography for cardiac output. ET-1 slightly increased mean arterial pressure and peripheral resistance but had no significant effect on systolic blood pressure and pulse pressure. PWV increased from 5.4+/-0.2 to 5.7+/-0.3 m/s (P<0.05), AIx from 9.9+/-3.3 to 17.2+/-3.8 (P<0.05), central systolic blood pressure by 8.7+/-1.7 mm Hg (P<0.05), and central pulse pressure by 5.1+/-1.9 mm Hg (P<0.05). This was associated with a fall in cardiac output by approximately 18% (P<0.05). VML-588 caused a slight decrease in brachial mean arterial pressure, PWV, and AIx, and prevented the effects of ET-1 on central hemodynamics without a clear dose-response effect. In summary, ET-1 in plasma concentrations as found in renal failure and heart failure accelerates PWV, causes a disproportionate increase in central aortic systolic blood pressure and pulse pressure, and decreases cardiac output. These effects can be prevented with an ET 1 receptor blocker such as VML-588. This makes it worthwhile to focus on endothelin as a target to prevent ventricular hypertrophy and to maintain cardiac function in diseases associated with high ET-1. PMID- 12743010 TI - Uric acid stimulates monocyte chemoattractant protein-1 production in vascular smooth muscle cells via mitogen-activated protein kinase and cyclooxygenase-2. AB - Previous studies have reported that uric acid stimulates vascular smooth muscle cell (VSMC) proliferation in vitro. We hypothesized that uric acid may also have direct proinflammatory effects on VSMCs. Crystal- and endotoxin-free uric acid was found to increase VSMC monocyte chemoattractant protein-1 (MCP-1) expression in a time- and dose-dependent manner, peaking at 24 hours. Increased mRNA and protein expression occurred as early as 3 hours after uric acid incubation and was partially dependent on posttranscriptional modification of MCP-1 mRNA. In addition, uric acid activated the transcription factors nuclear factor-kappaB and activator protein-1, as well as the MAPK signaling molecules ERK p44/42 and p38, and increased cyclooxygenase-2 (COX-2) mRNA expression. Inhibition of p38 (with SB 203580), ERK 44/42 (with UO126 or PD 98059), or COX-2 (with NS398) each significantly suppressed uric acid-induced MCP-1 expression at 24 hours, implicating these pathways in the response to uric acid. The ability of both n acetyl-cysteine and diphenyleneionium (antioxidants) to inhibit uric acid-induced MCP-1 production suggested involvement of intracellular redox pathways. Uric acid regulates critical proinflammatory pathways in VSMCs, suggesting it may have a role in the vascular changes associated with hypertension and vascular disease. PMID- 12743012 TI - Early autonomic dysfunction in glucose-tolerant but insulin-resistant offspring of type 2 diabetic patients. AB - In type 2 diabetes, both insulin resistance and hyperglycemia are considered responsible for autonomic dysfunction, but the specific role of these two abnormalities is not clear. To test the specific role of insulin resistance on autonomic dysfunction, we studied 69 glucose-tolerant offspring of type 2 diabetic patients, comparing the most insulin-resistant tertile (IR) with the most insulin-sensitive tertile (IS) and comparable control subjects, all undergoing the oral glucose tolerance test, impedentiometry, 24-hour blood pressure and ECG monitoring, and an intravenous glucose tolerance test (IVGTT) followed by a euglycemic hyperinsulinemic clamp, with continuous blood pressure and ECG measurements. Sympathovagal balance was evaluated as low- to high frequency ratio (LF:HF) by spectral analysis on R-R intervals. The change of systolic and diastolic blood pressure was calculated as [(day-night/d)]x100. In IR, the changes of systolic and diastolic blood pressure were significantly lower versus IS (9.2+/-5.0% versus 12.4+/-3.6%, P<0.02; 13.2+/-6.5% versus 17.4+/-5.2%, P<0.02). During the night, LF:HF fall was reduced in IR (43.1+/-21.0 versus 61.4+/-16.9, P<0.02). Hyperinsulinemia (IVGTT) rapidly and significantly increased LF:HF in IR (4.9+/-3.3 versus basal: 2.3+/-1.4, P=0.03) but not in IS. In offspring of type 2 diabetic patients with normal glucose tolerance and normal blood pressure values, insulin resistance is associated with abnormal control of blood pressure and sympathetic activation. Insulin resistance may therefore be responsible for some early derangements of the autonomic nervous tone control and thus contributes to increase the incidence of arterial hypertension and/or diabetes. PMID- 12743013 TI - Influence of vitamin C on baroreflex sensitivity in chronic heart failure. AB - Chronic heart failure (CHF) reduces baroreflex sensitivity. Low baroreflex sensitivity, a risk factor for sudden death, could arise partly from CHF dependent endothelial dysfunction. Vitamin C at high doses has a protective role against CHF-related endothelial damage. This study was conducted to investigate the effect of vitamin C on baroreflex sensitivity in CHF. A study group of 33 subjects with CHF secondary to postischemic dilated cardiomyopathy with an ejection fraction 0.20). However, concentrations of vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 (ICAM-1) in particular were higher in the PE group by 14% (P=0.038) and 44% (P=0.002), respectively. The cases also demonstrated a tendency toward higher fasting insulin (P=0.08) concentrations and had higher glycosylated hemoglobin levels (P=0.004). Leptin concentrations were not significantly elevated. Interestingly, significantly more of the women with history of PE were classified as menopausal (37.55% vs 17.5%, P=0.045). The differences in ICAM-1 concentration persisted (P=0.010) after adjustment for potential confounders, including hormonal use/menopausal status, antihypertensive or lipid-lowering therapy, and social class. We conclude that classic risk factors alone cannot fully explain the elevated CHD risk in women with a history of PE. Rather markedly elevated ICAM-1 concentrations and specific but subtle features of the metabolic syndrome (glucose, blood pressure) are likely to be involved. PMID- 12743017 TI - Phylogenetically older introns strongly correlate with module boundaries in ancient proteins. AB - The hypothesis that some (but not all) introns were used to construct ancient genes by exon shuffling of modules at the earliest stages of evolution is supported by the finding of an excess of phase-zero intron positions in the boundary regions of such modules in 276 ancient proteins (defined as common to eukaryotes and prokaryotes). Here we show further that as phase-zero intron positions are shared by distant taxa, and thus are truly phylogenetically ancient, their excess in the boundaries becomes greater, rising to an 80% excess if shared by four out of the five taxa: vertebrates, invertebrates, fungi, plants, and protists. PMID- 12743020 TI - Genome function and nuclear architecture: from gene expression to nanoscience. AB - Biophysical, chemical, and nanoscience approaches to the study of nuclear structure and activity have been developing recently and hold considerable promise. A selection of fundamental problems in genome organization and function are reviewed and discussed in the context of these new perspectives and approaches. Advancing these concepts will require coordinated networks of physicists, chemists, and materials scientists collaborating with cell, developmental, and genome biologists. PMID- 12743018 TI - Functional divergence of two zebrafish midkine growth factors following fish specific gene duplication. AB - In mammals, the unique midkine (mdk) gene encodes a secreted heparin-binding growth factor with neurotrophic activity. Here, we show the presence of two functional mdk genes named mdka and mdkb in zebrafish and rainbow trout. Both midkine proteins are clearly different from the related pleiotrophin, which was also identified in zebrafish and other fishes. Zebrafish mdka and mdkb genes map to linkage groups LG7 and LG25, respectively, both presenting synteny to human chromosome 11, in which the unique human ortholog mdk is located. At least four other genes unique in mammals are also present as duplicates on LG7 and LG25. Phylogenetic and divergence analyses suggested that LG7/LG25 paralogs including mdka and mdkb have been formed at approximately the same time, early during the evolution of the fish lineage. Hence, zebrafish and rainbow trout mdka and mdkb might have been generated by an ancient block duplication, and might be remnants of the proposed fish-specific whole-genome duplication. In contrast to the ubiquitous expression of their mammalian counterpart, zebrafish mdka and mdkb are expressed in spatially restricted, mostly nonoverlapping patterns during embryonic development and strongly in distinct domains in the adult brain. Ectopic ubiquitous expression of both mdk genes in early zebrafish embryos caused completely distinct effects on neural crest and floorplate development. These data indicate that mdka and mdkb underwent functional divergence after duplication. This provides an outstanding model to analyze the molecular mechanisms that lead to differences in pathways regulating the formation of homologous embryonic structures in different vertebrates. PMID- 12743019 TI - Assessment of SAGE in transcript identification. AB - An essential step in Serial Analysis of Gene Expression (SAGE) is tag mapping, which refers to the unambiguous determination of the gene represented by a SAGE tag. Current resources for tag mapping are incomplete, and thus do not allow assessment of the efficacy of SAGE in transcript identification. A method of tag mapping is described here and applied to the Drosophila melanogaster and Caenorhabditis elegans genomes, which permits detailed SAGE assessment and provides tag-mapping resources that were unavailable previously for these organisms. In our method, a conceptual transcriptome is constructed using genomic sequence and annotation by extending predicted coding regions to include UTRs on the basis of EST and cDNA alignments, UTR length distributions, and polyadenylation signals. Analysis of extracted tags suggests that, using the standard SAGE procedure, expression of 8% of D. melanogaster and 15% of C. elegans genes cannot be detected unambiguously by SAGE due to shared sequence or lack of NlaIII-anchoring enzyme sites. Both increasing tag length by 2-3 bp and using Sau3A instead of NlaIII as the anchoring enzyme increases potential for transcript detection. This work identifies and quantifies genes not amenable to SAGE analysis, in addition to providing tag-to-gene mappings for two model organisms. PMID- 12743021 TI - Differential expansion of zinc-finger transcription factor loci in homologous human and mouse gene clusters. AB - Mammalian genomes carry hundreds of Kruppel-type zinc finger (ZNF) genes, most of which reside in familial clusters. ZNF genes encoding Kruppel-associated box (KRAB) motifs are especially prone to this type of tandem organization. Despite their prevalence, little is known about the functions or evolutionary histories of these clustered gene families. Here we describe a homologous pair of human and mouse KRAB-ZNF gene clusters containing 21 human and 10 mouse genes, respectively. Evolutionary analysis uncovered only three pairs of putative orthologs and two cases where a single gene in one species is related to multiple genes in the other; several human genes have no obvious homolog in mouse. We deduce that duplication and loss of ancestral cluster members occurred independently in the primate and rodent lineages after divergence, yielding substantially different ZNF gene repertoires in humans and mice. Differences in expression patterns and sequence divergence within the DNA binding regions of predicted proteins suggest that the duplicated genes have acquired novel functions over evolutionary time. Since KRAB-ZNF proteins are predicted to function as transcriptional regulators, the elaboration of new lineage-specific genes in this and other clustered ZNF families is likely to have had a significant impact on species-specific aspects of biology. PMID- 12743022 TI - An appraisal of the potential for illegitimate recombination in bacterial genomes and its consequences: from duplications to genome reduction. AB - An exhaustive search for shortly spaced repeats in 74 bacterial chromosomes reveals that they are much more numerous than is usually acknowledged. These repeats were divided into five classes: close repeats (CRs), tandem repeats (TRs), simple sequence repeats (SSRs), spaced interspersed direct repeats, and "others." CRs are widespread and constitute the most abundant class, particularly in coding sequences. The other classes are less frequent, but each individual element shows a higher potential for recombination, when the number of repeats and their distances are taken into account. SSRs and TRs are more frequent in pathogens, as expected given their role in contingency loci, but are also widespread in the other bacteria. The analysis of CRs shows that they have an important role in the evolution of genomes, namely by generating duplications and deletions. Several cases compatible with a significant role of small CRs in the formation of large repeats were detected. Also, gene deletion in Buchnera correlates with repeat density, suggesting that CRs may lead to sequence deletion in general and genome reductive evolution of obligatory intracellular bacteria in particular. The assembly of these results indicates that shortly spaced repeats are key players in the dynamics of genome evolution. PMID- 12743023 TI - Comparative genome organization of human, murine, and feline MHC class II region. AB - To study comparative molecular dynamics in the genesis of the major histocompatibility complex (MHC), we determined a complete nucleotide sequence spanning 758,291 bp of the domestic cat (Felis catus) extended and classical class II region. The feline class II MHC includes 44 genes (31 predicted to be expressed) which display DNA sequence homology and ordered gene synteny with human HLA and mouse H2, in extended class II and centromere proximal regions (DM to DO) of the classical class II region. However, remarkable genomic alterations including gene gain and loss plus size differentials of 250 kb are evident in comparisons of the cat class II with those of human and mouse. The cat MHC lacks the entire DQ region and retains only relict pseudogene homologs of DP genes, compensated by expansion and reorganization of seven modern DR genes. Repetitive gene families within the feline MHC comprise 35% of the feline MHC with very different density and abundance of GC levels, SINES, LINES, STRs, and retro elements from the same repeats in human and mouse MHC. Comparison of the feline MHC with the murine and human MHC offers a detailed view of the consequences of genome organization in three mammalian lineages. PMID- 12743024 TI - Human-mouse gene identification by comparative evidence integration and evolutionary analysis. AB - The identification of genes in the human genome remains a challenge, as the actual predictions appear to disagree tremendously and vary dramatically on the basis of the specific gene-finding methodology used. Because the pattern of conservation in coding regions is expected to be different from intronic or intergenic regions, a comparative computational analysis can lead, in principle, to an improved computational identification of genes in the human genome by using a reference, such as mouse genome. However, this comparative methodology critically depends on three important factors: (1) the selection of the most appropriate reference genome. In particular, it is not clear whether the mouse is at the correct evolutionary distance from the human to provide sufficiently distinctive conservation levels in different genomic regions, (2) the selection of comparative features that provide the most benefit to gene recognition, and (3) the selection of evidence integration architecture that effectively interprets the comparative features. We address the first question by a novel evolutionary analysis that allows us to explicitly correlate the performance of the gene recognition system with the evolutionary distance (time) between the two genomes. Our simulation results indicate that there is a wide range of reference genomes at different evolutionary time points that appear to deliver reasonable comparative prediction of human genes. In particular, the evolutionary time between human and mouse generally falls in the region of good performance; however, better accuracy might be achieved with a reference genome further than mouse. To address the second question, we propose several natural comparative measures of conservation for identifying exons and exon boundaries. Finally, we experiment with Bayesian networks for the integration of comparative and compositional evidence. PMID- 12743025 TI - For whom the bell tolls: protein quality control of the endoplasmic reticulum and the ubiquitin-proteasome connection. AB - The surveillance of the structural fidelity of the proteome is of utmost importance to all cells. The endoplasmic reticulum (ER) is the organelle responsible for proper folding and delivery of proteins to the secretory pathway. It contains a sophisticated protein proofreading and elimination mechanism. Failure of this machinery leads to disease and, finally, to cell death. Elimination of misfolded proteins requires retrograde transport across the ER membrane and depends on the central cytoplasmic proteolytic machinery involved in cellular regulation: the ubiquitin-proteasome system. The basics of this process as well as recent advances in the field are reviewed. PMID- 12743026 TI - Changing neighbours, changing behaviour: cell adhesion molecule-mediated signalling during tumour progression. AB - Changes in cell-cell and cell-matrix adhesion accompany the transition from benign tumours to invasive, malignant cancer and the subsequent metastatic dissemination of tumour cells. This review discusses a possible role of cell adhesion molecules not only in redirecting a tumour cell's adhesive capabilities but also in modulating intracellular signalling, and with it, tumour malignancy. PMID- 12743028 TI - Transcarboxylase 12S crystal structure: hexamer assembly and substrate binding to a multienzyme core. AB - Transcarboxylase from Propionibacterium shermanii is a 1.2 MDa multienzyme complex that couples two carboxylation reactions, transferring CO(2)(-) from methylmalonyl-CoA to pyruvate, yielding propionyl-CoA and oxaloacetate. The 1.9 A resolution crystal structure of the central 12S hexameric core, which catalyzes the first carboxylation reaction, has been solved bound to its substrate methylmalonyl-CoA. Overall, the structure reveals two stacked trimers related by 2-fold symmetry, and a domain duplication in the monomer. In the active site, the labile carboxylate group of methylmalonyl-CoA is stabilized by interaction with the N-termini of two alpha-helices. The 12S domains are structurally similar to the crotonase/isomerase superfamily, although only domain 1 of each 12S monomer binds ligand. The 12S reaction is similar to that of human propionyl-CoA carboxylase, whose beta-subunit has 50% sequence identity with 12S. A homology model of the propionyl-CoA carboxylase beta-subunit, based on this 12S crystal structure, provides new insight into the propionyl-CoA carboxylase mechanism, its oligomeric structure and the molecular basis of mutations responsible for enzyme deficiency in propionic acidemia. PMID- 12743027 TI - The integrin-actin connection, an eternal love affair. AB - Integrin receptors connect the extracellular matrix to the actin cytoskeleton. This interaction can be viewed as a cyclical liaison, which develops again and again at new adhesion sites only to cease at sites of de-adhesion. Recent work has demonstrated that multidomain proteins play crucial roles in the integrin actin connection by providing a high degree of regulation adjusted to the needs of the cell. In this review we present several examples of this paradigm and with special emphasis on the ILK-PINCH-parvin complex, which amply demonstrates how structural and signalling functions are linked together. PMID- 12743029 TI - Crystal structure of the CUB1-EGF-CUB2 region of mannose-binding protein associated serine protease-2. AB - Serum mannose-binding proteins (MBPs) are C-type lectins that recognize cell surface carbohydrate structures on pathogens, and trigger killing of these targets by activating the complement pathway. MBPs circulate as a complex with MBP-associated serine proteases (MASPs), which become activated upon engagement of a target cell surface. The minimal functional unit for complement activation is a MASP homodimer bound to two MBP trimeric subunits. MASPs have a modular structure consisting of an N-terminal CUB domain, a Ca(2+)-binding EGF-like domain, a second CUB domain, two complement control protein modules and a C terminal serine protease domain. The CUB1-EGF-CUB2 region mediates homodimerization and binding to MBP. The crystal structure of the MASP-2 CUB1-EGF CUB2 dimer reveals an elongated structure with a prominent concave surface that is proposed to be the MBP-binding site. A model of the full six-domain structure and its interaction with MBPs suggests mechanisms by which binding to a target cell transmits conformational changes from MBP to MASP that allow activation of its protease activity. PMID- 12743030 TI - Proteolytic processing is necessary to separate and ensure proper cell growth and cytokinesis functions of HCF-1. AB - HCF-1 is a highly conserved and abundant chromatin-associated host cell factor required for transcriptional activation of herpes simplex virus immediate-early genes by the virion protein VP16. HCF-1 exists as a heterodimeric complex of associated N- (HCF-1(N)) and C- (HCF-1(C)) terminal subunits that result from proteolytic processing of a precursor protein. We have used small-interfering RNA (siRNA) to inactivate HCF-1 in an array of normal and transformed mammalian cells to identify its cellular functions. Our results show that HCF-1 is a broadly acting regulator of two stages of the cell cycle: exit from mitosis, where it ensures proper cytokinesis, and passage through the G(1) phase, where it promotes cell cycle progression. Proteolytic processing is necessary to separate and ensure these two HCF-1 activities, which are performed by separate HCF-1 subunits: the HCF-1(N) subunit promotes passage through the G(1) phase whereas the HCF-1(C) subunit is involved in proper exit from mitosis. These results suggest that HCF-1 links the regulation of exit from mitosis and the G(1) phase of cell growth, possibly to coordinate the reactivation of gene expression after mitosis. PMID- 12743031 TI - A permease-like protein involved in ER to thylakoid lipid transfer in Arabidopsis. AB - In eukaryotes, enzymes of different subcellular compartments participate in the assembly of membrane lipids. As a consequence, interorganelle lipid transfer is extensive in growing cells. A prominent example is the transfer of membrane lipid precursors between the endoplasmic reticulum (ER) and the photosynthetic thylakoid membranes in plants. Mono- and digalactolipids are typical photosynthetic membrane lipids. In Arabidopsis, they are derived from one of two pathways, either synthesized de novo in the plastid, or precursors are imported from the ER, giving rise to distinct molecular species. Employing a high throughput robotic screening procedure generating arrays of spot chromatograms, mutants of Arabidopsis were isolated, which accumulated unusual trigalactolipids. In one allelic mutant subclass, trigalactosyldiacylglycerol1, the primary defect caused a disruption in the biosynthesis of ER-derived thylakoid lipids. Secondarily, a processive galactosyltransferase was activated, leading to the accumulation of oligogalactolipids. Mutations in a permease-like protein of the outer chloroplastic envelope are responsible for the primary biochemical defect. It is proposed that this protein is part of a lipid transfer complex. PMID- 12743032 TI - Tom40, the import channel of the mitochondrial outer membrane, plays an active role in sorting imported proteins. AB - The Translocase of the Outer Mitochondrial membrane (TOM complex) is centred on a channel, created by Tom40, serving as the only means of entry for proteins into the mitochondrion. Proteins destined for internal mitochondrial compartments interact subsequently with one of the two distinct protein Translocases of the Inner Mitochondrial membrane (TIM23 and TIM54 complexes) or follow specialized paths into the intermembrane space. To investigate the sorting of precursor proteins to these various sub-mitochondrial compartments, we created a library of tom40 mutants and screened for alleles selectively corrupt in protein sorting. One of the tom40 mutants, tom40-97, carries a single point mutation (W(243)R) resulting in an ineffective transfer of precursors to the TIM23 complex. There is no defect on transfer of precursors to the TIM54 complex or insertion of proteins into the outer membrane. The Tom40 channel is not a passive pore, but plays an active role in protein sorting for all sub-mitochondrial locations. PMID- 12743033 TI - Conventional kinesin KIF5B mediates insulin-stimulated GLUT4 movements on microtubules. AB - Insulin stimulates glucose uptake in muscle and adipose cells by mobilizing intracellular membrane vesicles containing GLUT4 glucose transporter proteins to the plasma membrane. Here we show in live cultured adipocytes that intracellular membranes containing GLUT4-yellow fluorescent protein (YFP) move along tubulin cyan fluorescent protein-labeled microtubules in response to insulin by a mechanism that is insensitive to the phosphatidylinositol 3 (PI3)-kinase inhibitor wortmannin. Insulin increased by several fold the observed frequencies, but not velocities, of long-range movements of GLUT4-YFP on microtubules, both away from and towards the perinuclear region. Genomics screens show conventional kinesin KIF5B is highly expressed in adipocytes and this kinesin is partially co localized with perinuclear GLUT4. Dominant-negative mutants of conventional kinesin light chain blocked outward GLUT4 vesicle movements and translocation of exofacial Myc-tagged GLUT4-green fluorescent protein to the plasma membrane in response to insulin. These data reveal that insulin signaling targets the engagement or initiates the movement of GLUT4-containing membranes on microtubules via conventional kinesin through a PI3-kinase-independent mechanism. This insulin signaling pathway regulating KIF5B function appears to be required for GLUT4 translocation to the plasma membrane. PMID- 12743034 TI - Laminin-10 is crucial for hair morphogenesis. AB - The role of the extracellular matrix in cutaneous morphogenesis is poorly understood. Here, we describe the essential role of laminin-10 (alpha5beta1gamma1) in hair follicle development. Laminin-10 was present in the basement membrane of elongating hair germs, when other laminins were downregulated, suggesting a role for laminin-10 in hair development. Treatment of human scalp xenografts with antibodies to laminin-10, or its receptor beta1 integrin, produced alopecia. E16.5 Lama5 -/- mouse skin, lacking laminin-10, contained fewer hair germs compared with controls, and after transplantation, Lama5 -/- skin showed a failure of hair germ elongation followed by complete hair follicle regression. Lama5 -/- skin showed defective basement membrane assembly, without measurable increases in anoikis. Instead, Lama5 -/- skin showed decreased expression of early hair markers including sonic hedgehog and Gli1, implicating laminin-10 in developmental signaling. Intriguingly, treatment of Lama5 -/- skin with purified laminin-10 corrected basement membrane defects and restored hair follicle development. We conclude that laminin-10 is required for hair follicle development and report the first use of exogenous protein to correct a cutaneous developmental defect. PMID- 12743035 TI - TACE cleavage of proamphiregulin regulates GPCR-induced proliferation and motility of cancer cells. AB - Communication between G protein-coupled receptor (GPCR) and epidermal growth factor receptor (EGFR) signalling systems involves cell surface proteolysis of EGF-like precursors. The underlying mechanisms of EGFR signal transactivation pathways, however, are largely unknown. We demonstrate that in squamous cell carcinoma cells, stimulation with the GPCR agonists LPA or carbachol specifically results in metalloprotease cleavage and release of amphiregulin (AR). Moreover, AR gene silencing by siRNA or inhibition of AR biological activity by neutralizing antibodies and heparin prevents GPCR-induced EGFR tyrosine phosphorylation, downstream mitogenic signalling events, cell proliferation, migration and activation of the survival mediator Akt/PKB. Therefore, despite some functional redundancy among EGF family ligands, the present study reveals a distinct and essential role for AR in GPCR-triggered cellular responses. Furthermore, we present evidence that blockade of the metalloprotease-disintegrin tumour necrosis factor-alpha-converting enzyme (TACE) by the tissue inhibitor of metalloprotease-3, a dominant-negative TACE mutant or RNA interference suppresses GPCR-stimulated AR release, EGFR activation and downstream events. Thus, TACE can function as an effector of GPCR-mediated signalling and represents a key element of the cellular receptor cross-talk network. PMID- 12743036 TI - The mitogen-activated protein (MAP) kinase ERK induces tRNA synthesis by phosphorylating TFIIIB. AB - RNA polymerase (pol) III transcription increases within minutes of serum addition to growth-arrested fibroblasts. We show that ERK mitogen-activated protein kinases regulate pol III output by directly binding and phosphorylating the BRF1 subunit of transcription factor TFIIIB. Blocking the ERK signalling cascade inhibits TFIIIB binding to pol III and to transcription factor TFIIIC2. Chromatin immunoprecipitation shows that the association of BRF1 and pol III with tRNA(Leu) genes in cells decreases when ERK is inactivated. Furthermore, mutation of an ERK docking domain or phosphoacceptor site in BRF1 prevents serum induction of pol III transcription. These data identify a novel target for ERK, and suggest that its ability to stimulate biosynthetic capacity and growth involves direct transcriptional activation of tRNA and 5S rRNA genes. PMID- 12743037 TI - Osmostress-induced transcription by Hot1 depends on a Hog1-mediated recruitment of the RNA Pol II. AB - In budding yeast, the mitogen-activated protein kinase (MAPK) Hog1 coordinates the transcriptional program required for cell survival upon osmostress. The Hot1 transcription factor acts downstream of the MAPK and regulates a subset of Hog1 responsive genes. In response to high osmolarity, Hot1 targets Hog1 to specific osmostress-responsive promoters. Here, we show that assembly of the general transcription machinery at Hot1-dependent promoters depends on the presence of Hot1 and active Hog1 MAPK. Unexpectedly, recruitment of RNA polymerase (Pol) II complex to target promoters does not depend on the phosphorylation of the Hot1 activator by the MAPK. Hog1 interacts with the RNA Pol II and with general components of the transcription machinery. More over, when tethered to a promoter as a LexA fusion protein, Hog1 activates transcription in a stress- regulated manner. Thus, anchoring of active Hog1 to promoters by the Hot1 activator is essential for recruitment and activation of RNA Pol II. The mammalian p38 also interacts with the RNA Pol II, which might suggest a conserved mechanism for regulation of gene expression by SAPKs among eukaryotic cells. PMID- 12743038 TI - Opposing functions of ZEB proteins in the regulation of the TGFbeta/BMP signaling pathway. AB - Binding of TGFbeta/BMP factors to their receptors leads to translocation of Smad proteins to the nucleus where they activate transcription of target genes. The two-handed zinc finger proteins encoded by Zfhx1a and Zfhx1b, ZEB-1/deltaEF1 and ZEB-2/SIP1, respectively, regulate gene expression and differentiation programs in a number of tissues. Here I demonstrate that ZEB proteins are also crucial regulators of TGFbeta/BMP signaling with opposing effects on this pathway. Both ZEB proteins bind to Smads, but while ZEB-1/deltaEF1 synergizes with Smad proteins to activate transcription, promote osteoblastic differentiation and induce cell growth arrest, the highly related ZEB-2/SIP1 protein has the opposite effect. Finally, the ability of TGFbeta to mediate transcription of TGFbeta dependent genes and induce growth arrest depends on the presence of endogenous ZEB-1/deltaEF1 protein. PMID- 12743039 TI - Regulation of Smad signaling through a differential recruitment of coactivators and corepressors by ZEB proteins. AB - Balancing signals derived from the TGFbeta family is crucial for regulating cell proliferation and differentiation, and in establishing the embryonic axis during development. TGFbeta/BMP signaling leads to the activation and nuclear translocation of Smad proteins, which activate transcription of specific target genes by recruiting P/CAF and p300. The two members of the ZEB family of zinc finger factors (ZEB-1/deltaEF1 and ZEB-2/SIP1) regulate TGFbeta/BMP signaling in opposite ways: ZEB-1/deltaEF1 synergizes with Smad-mediated transcriptional activation, while ZEB-2/SIP1 represses it. Here we report that these antagonistic effects by the ZEB proteins arise from the differential recruitment of transcriptional coactivators (p300 and P/CAF) and corepressors (CtBP) to the Smads. Thus, while ZEB-1/deltaEF1 binds to p300 and promotes the formation of a p300-Smad transcriptional complex, ZEB-2/SIP1 acts as a repressor by recruiting CtBP. This model of regulation by ZEB proteins also functions in vivo, where they have opposing effects on the regulation of TGFbeta family-dependent genes during Xenopus development. PMID- 12743041 TI - Genome-wide analysis of nuclear mRNA export pathways in Drosophila. AB - NXF1, p15 and UAP56 are essential nuclear mRNA export factors. The fraction of mRNAs exported by these proteins or via alternative pathways is unknown. We have analyzed the relative abundance of nearly half of the Drosophila transcriptome in the cytoplasm of cells treated with the CRM1 inhibitor leptomycin-B (LMB) or depleted of export factors by RNA interference. While the vast majority of mRNAs were unaffected by LMB, the levels of most mRNAs were significantly reduced in cells depleted of NXF1, p15 or UAP56. The striking similarities of the mRNA expression profiles in NXF1, p15 and UAP56 knockdowns show that these proteins act in the same pathway. The broad effect on mRNA levels observed in these cells indicates that the functioning of this pathway is required for export of most mRNAs. Nonetheless, a set of mRNAs whose export was unaffected by the depletions and some requiring NXF1:p15 but not UAP56 were identified. In addition, our analysis revealed a feedback loop by which a block to mRNA export triggers the upregulation of genes involved in this process. PMID- 12743042 TI - The Drosophila hairy RNA localization signal modulates the kinetics of cytoplasmic mRNA transport. AB - In several Drosophila cell types, mRNA transport depends on microtubules, the molecular motor dynein and trans-acting factors including Egalitarian and Bicaudal-D. However, the molecular basis of transcript recognition by the localization machinery is poorly understood. Here, we characterize the features of hairy pair-rule RNA transcripts that mediate their apical localization, using in vivo injection of fluorescently labelled mRNAs into syncytial blastoderm embryos. We show that a 121-nucleotide element within the 3'-untranslated region is necessary and sufficient to mediate apical transport. The signal comprises two essential stem-loop structures, in which double-stranded stems are crucial for localization. Base-pair identities within the stems are not essential, but can contribute to the efficiency of localization, suggesting that specificity is mediated by higher-order structure. Using time-lapse microscopy, we measure the kinetics of localization and show that impaired localization of mutant signals is due to delayed formation of active motor complexes and, unexpectedly, to slower movement. These findings, and those from co-injecting wild-type and mutant RNAs, suggest that the efficiency of molecular motors is modulated by the character of their cargoes. PMID- 12743040 TI - A test of insulator interactions in Drosophila. AB - Insulators are a class of elements that define independent domains of gene function. The Drosophila gypsy insulator is proposed to establish regulatory isolation by forming loop domains that constrain interactions between transcriptional control elements. This supposition is based upon the observation that insertion of a single gypsy insulator between an enhancer and promoter blocks enhancer function, while insertion of two gypsy insulators promotes enhancer bypass and activation of transcription. To investigate this model, we determined whether non-gypsy insulators interacted with each other and with the gypsy insulator. Pairs of scs or scs' insulators blocked enhancer function. Further, an intervening scs insulator did not block gypsy insulator interactions. Taken together, these data suggest that not all Drosophila insulators interact, with this property restricted to some insulators, such as gypsy. Three gypsy insulators inserted between an enhancer and promoter blocked enhancer function, indicating that gypsy insulator interactions may be restricted to pairs. Our studies imply that formation of loop domains may represent one of many mechanisms used by insulators to impart regulatory isolation. PMID- 12743044 TI - Completion of a parasexual cycle in Candida albicans by induced chromosome loss in tetraploid strains. AB - The human pathogenic fungus Candida albicans has traditionally been classified as a diploid, asexual organism. However, mating-competent forms of the organism were recently described that produced tetraploid mating products. In principle, the C.albicans life cycle could be completed via a sexual process, via a parasexual mechanism, or by both mechanisms. Here we describe conditions in which growth of a tetraploid strain of C.albicans on Saccharomyces cerevisiae 'pre-sporulation' medium induced efficient, random chromosome loss in the tetraploid. The products of chromosome loss were often strains that were diploid, or very close to diploid, in DNA content. If they inherited the appropriate MTL (mating-type like) loci, these diploid products were themselves mating competent. Thus, an efficient parasexual cycle can be performed in C.albicans, one that leads to the reassortment of genetic material in this organism. We show that this parasexual cycle-consisting of mating followed by chromosome loss-can be used in the laboratory for simple genetic manipulations in C.albicans. PMID- 12743043 TI - Centromeric chromatin pliability and memory at a human neocentromere. AB - We show that Trichostatin A (TSA)-induced partial histone hyperacetylation causes a unidirectional shift in the position of a previously defined binding domain for the centromere-specific histone H3 homologue CENP-A at a human neocentromere. The shift of approximately 320 kb is fully reversible when TSA is removed, but is accompanied by an apparent reduction in the density of CENP-A per unit length of genomic DNA at the neocentromere. TSA treatment also instigates a reversible abolition of a previously defined major domain of differentially delayed replication timing that was originally established at the neocentromeric site. None of these changes has any measurable deleterious effects on mitosis or neocentromere function. The data suggest pliability of centromeric chromatin in response to epigenetic triggers, and the non-essential nature of the regions of delayed replication for centromere function. Reversibility of the CENP-A-binding position and the predominant region of delayed replication timing following removal of TSA suggest strong memory at the original site of neocentromeric chromatin formation. PMID- 12743045 TI - A novel type of replicative enzyme harbouring ATPase, primase and DNA polymerase activity. AB - Although DNA replication is a process common in all domains of life, primase and replicative DNA polymerase appear to have evolved independently in the bacterial domain versus the archaeal/eukaryal branch of life. Here, we report on a new type of replication protein that constitutes the first member of the DNA polymerase family E. The protein ORF904, encoded by the plasmid pRN1 from the thermoacidophile archaeon Sulfolobus islandicus, is a highly compact multifunctional enzyme with ATPase, primase and DNA polymerase activity. Recombinant purified ORF904 hydrolyses ATP in a DNA-dependent manner. Deoxynucleotides are preferentially used for the synthesis of primers approximately 8 nucleotides long. The DNA polymerase activity of ORF904 synthesizes replication products of up to several thousand nucleotides in length. The primase and DNA polymerase activity are located in the N-terminal half of the protein, which does not show homology to any known DNA polymerase or primase. ORF904 constitutes a new type of replication enzyme, which could have evolved independently from the eubacterial and archaeal/eukaryal proteins of DNA replication. PMID- 12743047 TI - SARS: the struggle for containment. PMID- 12743052 TI - Requiem for a journal. PMID- 12743046 TI - Xenopus Cut5 is essential for a CDK-dependent process in the initiation of DNA replication. AB - Fission yeast Cut5/Rad4 and its budding yeast homolog Dpb11 are required for both DNA replication and the S-phase checkpoint. Here, we have investigated the role of the Xenopus homolog of Cut5 in the initiation of DNA replication using Xenopus egg extracts. Xenopus Cut5, which shows sequence similarity to DmMus101 and HsTopBP1, is essential for DNA replication in the egg extracts. It is required for the chromatin binding of Cdc45 and DNA polymerases, but not for the formation of pre-replicative complexes or the elongation stage of DNA replication. The chromatin binding of Cut5 consists of two distinct modes. S-phase cyclin dependent kinase (S-CDK)-independent binding is sufficient for DNA replication while S-CDK-dependent binding is dispensable. Further, S-CDK acts after the chromatin binding of Cut5 and before the binding of Cdc45. These results demonstrate that the chromatin binding of Cut5 is required for the action of S CDK, which in turn triggers the formation of pre-initiation complexes of DNA replication. PMID- 12743055 TI - Marketing Rx&D. PMID- 12743056 TI - Debating gun registration. PMID- 12743058 TI - Debating gun registration. PMID- 12743059 TI - Debating gun registration. PMID- 12743060 TI - Debating gun registration. PMID- 12743061 TI - Debating gun registration. PMID- 12743062 TI - Debating gun registration. PMID- 12743063 TI - Debating gun registration. PMID- 12743064 TI - Debating gun registration. PMID- 12743065 TI - The immediate psychological and occupational impact of the 2003 SARS outbreak in a teaching hospital. AB - BACKGROUND: The outbreak of severe acute respiratory syndrome (SARS) in Toronto, which began on Mar. 7, 2003, resulted in extraordinary public health and infection control measures. We aimed to describe the psychological and occupational impact of this event within a large hospital in the first 4 weeks of the outbreak and the subsequent administrative and mental health response. METHODS: Two principal authors met with core team members and mental health care providers at Mount Sinai Hospital, Toronto, to compile retrospectively descriptions of the experiences of staff and patients based on informal observation. All authors reviewed and analyzed the descriptions in an iterative process between Apr. 3 and Apr. 13, 2003. RESULTS: In a 4-week period, 19 individuals developed SARS, including 11 health care workers. The hospital's response included establishing a leadership command team and a SARS isolation unit, implementing mental health support interventions for patients and staff, overcoming problems with logistics and communication, and overcoming resistance to directives. Patients with SARS reported fear, loneliness, boredom and anger, and they worried about the effects of quarantine and contagion on family members and friends. They experienced anxiety about fever and the effects of insomnia. Staff were adversely affected by fear of contagion and of infecting family, friends and colleagues. Caring for health care workers as patients and colleagues was emotionally difficult. Uncertainty and stigmatization were prominent themes for both staff and patients. INTERPRETATION: The hospital's response required clear communication, sensitivity to individual responses to stress, collaboration between disciplines, authoritative leadership and provision of relevant support. The emotional and behavioural reactions of patients and staff are understood to be a normal, adaptive response to stress in the face of an overwhelming event. PMID- 12743067 TI - Psychological implications of abortion--highly charged and rife with misleading research. PMID- 12743068 TI - Containing a new infection with new technology: a Web-based response to SARS. PMID- 12743066 TI - Psychiatric admissions of low-income women following abortion and childbirth. AB - BACKGROUND: Controversy exists about whether abortion or childbirth is associated with greater psychological risks. We compared psychiatric admission rates of women in time periods from 90 days to 4 years after either abortion or childbirth. METHODS: We used California Medicaid (Medi-Cal) records of women aged 13-49 years at the time of either abortion or childbirth during 1989. Only women who had no psychiatric admissions or pregnancy events during the year before the target pregnancy event were included (n = 56 741). Psychiatric admissions were examined using logistic regression analyses, controlling for age and months of eligibility for Medi-Cal services. RESULTS: Overall, women who had had an abortion had a significantly higher relative risk of psychiatric admission compared with women who had delivered for every time period examined. Significant differences by major diagnostic categories were found for adjustment reactions (odds ratio [OR] 2.1, 95% confidence interval [CI] 1.1-4.1), single-episode (OR 1.9, 95% CI 1.3-2.9) and recurrent depressive psychosis (OR 2.1, 95% CI 1.3-3.5), and bipolar disorder (OR 3.0, 95% CI 1.5-6.0). Significant differences were also observed when the results were stratified by age. INTERPRETATION: Subsequent psychiatric admissions are more common among low-income women who have an induced abortion than among those who carry a pregnancy to term, both in the short and longer term. PMID- 12743069 TI - Global solidarity against globalized tobacco: the world's first modern health treaty tackles tobacco. PMID- 12743070 TI - The race to outpace severe acute respiratory syndrome (SARS). PMID- 12743072 TI - Marketing Rx&D: one step forward, two steps back. PMID- 12743071 TI - Getting it right: industry sponsorship and medical research. PMID- 12743076 TI - Ribavirin in the treatment of SARS: A new trick for an old drug? PMID- 12743077 TI - SARS update. PMID- 12743075 TI - Peanut allergy: an overview. AB - Peanut allergy accounts for the majority of severe food-related allergic reactions. It tends to present early in life, and affected individuals generally do not outgrow it. In highly sensitized people, trace quantities can induce an allergic reaction. In this review, we will discuss the prevalence, clinical characteristics, diagnosis, natural history and management of peanut allergy. PMID- 12743084 TI - MDs mostly mum on Iraq. PMID- 12743078 TI - Treatment of chronic obstructive pulmonary disease: Combination or component therapy? PMID- 12743085 TI - SARS: "a domino effect through entire system". PMID- 12743091 TI - For Iraqi women, more war means more uncertainty. PMID- 12743093 TI - Trends in post-MD training. PMID- 12743098 TI - Computational cell biology. PMID- 12743097 TI - Amyloid as a natural product. AB - Amyloid fibrils, such as those found in Alzheimer's and the gelsolin amyloid diseases, result from the misassembly of peptides produced by either normal or aberrant intracellular proteolytic processing. A paper in this issue by Marks and colleagues (Berson et al., 2003) demonstrates that intra-melanosome fibrils are formed through normal biological proteolytic processing of an integral membrane protein. The resulting peptide fragment assembles into fibrils promoting the formation of melanin pigment granules. These results, along with the observation that amyloid fibril formation by bacteria is highly orchestrated, suggest that fibril formation is an evolutionary conserved biological pathway used to generate natural product nanostructures. PMID- 12743096 TI - Holey barrier: claudins and the regulation of brain endothelial permeability. AB - Endothelial tight junctions (TJs)* are an important functional part of the blood brain barrier (BBB). In this issue, Nitta et al. (2003) demonstrate that claudin 5, a transmembrane protein of TJs, is a critical determinant of BBB permeability in mice. Unexpectedly, knockout of claudin-5 did not result in a general breakdown of TJs but in a selective increase in paracellular permeability of small molecules. This suggests that the BBB can be manipulated to allow selective diffusion of small molecules and makes claudin-5 a possible target for the development of drugs for this purpose. PMID- 12743099 TI - Inventories to insights. AB - "In the long course of cell life on this earth it remained, for our age, for our generation, to receive the full ownership of our inheritance. We have entered the cell, the Mansion of our birth and started the inventory of our acquired wealth." (Albert Claude, Nobel lecture, 1974). PMID- 12743100 TI - Modeling network dynamics: the lac operon, a case study. AB - We use the lac operon in Escherichia coli as a prototype system to illustrate the current state, applicability, and limitations of modeling the dynamics of cellular networks. We integrate three different levels of description (molecular, cellular, and that of cell population) into a single model, which seems to capture many experimental aspects of the system. PMID- 12743101 TI - Computational imaging in cell biology. AB - Microscopy of cells has changed dramatically since its early days in the mid seventeenth century. Image analysis has concurrently evolved from measurements of hand drawings and still photographs to computational methods that (semi-) automatically quantify objects, distances, concentrations, and velocities of cells and subcellular structures. Today's imaging technologies generate a wealth of data that requires visualization and multi-dimensional and quantitative image analysis as prerequisites to turning qualitative data into quantitative values. Such quantitative data provide the basis for mathematical modeling of protein kinetics and biochemical signaling networks that, in turn, open the way toward a quantitative view of cell biology. Here, we will review technologies for analyzing and reconstructing dynamic structures and processes in the living cell. We will present live-cell studies that would have been impossible without computational imaging. These applications illustrate the potential of computational imaging to enhance our knowledge of the dynamics of cellular structures and processes. PMID- 12743102 TI - Spindle orientation in Saccharomyces cerevisiae depends on the transport of microtubule ends along polarized actin cables. AB - Microtubules and actin filaments interact and cooperate in many processes in eukaryotic cells, but the functional implications of such interactions are not well understood. In the yeast Saccharomyces cerevisiae, both cytoplasmic microtubules and actin filaments are needed for spindle orientation. In addition, this process requires the type V myosin protein Myo2, the microtubule end-binding protein Bim1, and Kar9. Here, we show that fusing Bim1 to the tail of the Myo2 is sufficient to orient spindles in the absence of Kar9, suggesting that the role of Kar9 is to link Myo2 to Bim1. In addition, we show that Myo2 localizes to the plus ends of cytoplasmic microtubules, and that the rate of movement of these cytoplasmic microtubules to the bud neck depends on the intrinsic velocity of Myo2 along actin filaments. These results support a model for spindle orientation in which a Myo2-Kar9-Bim1 complex transports microtubule ends along polarized actin cables. We also present data suggesting that a similar process plays a role in orienting cytoplasmic microtubules in mating yeast cells. PMID- 12743103 TI - Neurofilament heavy chain side arm phosphorylation regulates axonal transport of neurofilaments. AB - Neurofilaments possess side arms that comprise the carboxy-terminal domains of neurofilament middle and heavy chains (NFM and NFH); that of NFH is heavily phosphorylated in axons. Here, we demonstrate that phosphorylation of NFH side arms is a mechanism for regulating transport of neurofilaments through axons. Mutants in which known NFH phosphorylation sites were mutated to preclude phosphorylation or mimic permanent phosphorylation display altered rates of transport in a bulk transport assay. Similarly, application of roscovitine, an inhibitor of the NFH side arm kinase Cdk5/p35, accelerates neurofilament transport. Analyses of neurofilament movement in transfected living neurons demonstrated that a mutant mimicking permanent phosphorylation spent a higher proportion of time pausing than one that could not be phosphorylated. Thus, phosphorylation of NFH slows neurofilament transport, and this is due to increased pausing in neurofilament movement. PMID- 12743104 TI - Androgen-stimulated DNA synthesis and cytoskeletal changes in fibroblasts by a nontranscriptional receptor action. AB - In NIH3T3 cells, 0.001 nM of the synthetic androgen R1881 induces and stimulates association of androgen receptor (AR) with Src and phosphatidylinositol 3-kinase (Pl3-kinase), respectively, thereby triggering S-phase entry. 10 nM R1881 stimulates Rac activity and membrane ruffling in the absence of the receptor-Src PI3-kinase complex assembly. The antiandrogen Casodex and specific inhibitors of Src and PI3-kinase prevent both hormonal effects, DNA synthesis and cytoskeletal changes. Neither low nor high R1881 concentration allows receptor nuclear translocation and receptor-dependent transcriptional activity in fibroblasts, although they harbor the classical murine AR. The very low amount of AR in NIH3T3 cells (7% of that present in LNCaP cells) activates the signaling pathways, but apparently is not sufficient to stimulate gene transcription. This view is supported by the appearance of receptor nuclear translocation as well as receptor mediated transcriptional activity after overexpression of AR in fibroblasts. In addition, AR-negative Cos cells transiently transfected with a very low amount of hAR cDNA respond to low and high R1881 concentrations with signaling activation. Interestingly, they do not show significant transcriptional activation under the same experimental conditions. Fibroblasts are the first example of cells that respond to steroid hormones with activation of signaling pathways in the absence of endogenous receptor transcriptional activity. The data reported also show that hormone concentration can be crucial in determining the type of cell responsiveness. PMID- 12743105 TI - Alpha-adducin dissociates from F-actin and spectrin during platelet activation. AB - Aspectrin-based skeleton uniformly underlies and supports the plasma membrane of the resting platelet, but remodels and centralizes in the activated platelet. alpha-Adducin, a phosphoprotein that forms a ternary complex with F-actin and spectrin, is dephosphorylated and mostly bound to spectrin in the membrane skeleton of the resting platelet at sites where actin filaments attach to the ends of spectrin molecules. Platelets activated through protease-activated receptor 1, FcgammaRIIA, or by treatment with PMA phosphorylate adducin at Ser726. Phosphoadducin releases from the membrane skeleton concomitant with its dissociation from spectrin and actin. Inhibition of PKC blunts adducin phosphorylation and release from spectrin and actin, preventing the centralization of spectrin that normally follows cell activation. We conclude that adducin targets actin filament ends to spectrin to complete the assembly of the resting membrane skeleton. Dissociation of phosphoadducin releases spectrin from actin, facilitating centralization of spectrin, and leads to the exposure of barbed actin filament ends that may then participate in converting the resting platelet's disc shape into its active form. PMID- 12743106 TI - Intact satellite cells lead to remarkable protection against Smn gene defect in differentiated skeletal muscle. AB - Deletion of murine Smn exon 7, the most frequent mutation found in spinal muscular atrophy, has been directed to either both satellite cells, the muscle progenitor cells and fused myotubes, or fused myotubes only. When satellite cells were mutated, mutant mice develop severe myopathic process, progressive motor paralysis, and early death at 1 mo of age (severe mutant). Impaired muscle regeneration of severe mutants correlated with defect of myogenic precursor cells both in vitro and in vivo. In contrast, when satellite cells remained intact, mutant mice develop similar myopathic process but exhibit mild phenotype with median survival of 8 mo and motor performance similar to that of controls (mild mutant). High proportion of regenerating myofibers expressing SMN was observed in mild mutants compensating for progressive loss of mature myofibers within the first 6 mo of age. Then, in spite of normal contractile properties of myofibers, mild mutants develop reduction of muscle force and mass. Progressive decline of muscle regeneration process was no more able to counterbalance muscle degeneration leading to dramatic loss of myofibers. These data indicate that intact satellite cells remarkably improve the survival and motor performance of mutant mice suffering from chronic myopathy, and suggest a limited potential of satellite cells to regenerate skeletal muscle. PMID- 12743107 TI - deep-orange and carnation define distinct stages in late endosomal biogenesis in Drosophila melanogaster. AB - Endosomal degradation is severely impaired in primary hemocytes from larvae of eye color mutants of Drosophila. Using high resolution imaging and immunofluorescence microscopy in these cells, products of eye color genes, deep orange (dor) and carnation (car), are localized to large multivesicular Rab7 positive late endosomes containing Golgi-derived enzymes. These structures mature into small sized Dor-negative, Car-positive structures, which subsequently fuse to form tubular lysosomes. Defective endosomal degradation in mutant alleles of dor results from a failure of Golgi-derived vesicles to fuse with morphologically arrested Rab7-positive large sized endosomes, which are, however, normally acidified and mature with wild-type kinetics. This locates the site of Dor function to fusion of Golgi-derived vesicles with the large Rab7-positive endocytic compartments. In contrast, endosomal degradation is not considerably affected in car1 mutant; fusion of Golgi-derived vesicles and maturation of large sized endosomes is normal. However, removal of Dor from small sized Car-positive endosomes is slowed, and subsequent fusion with tubular lysosomes is abolished. Overexpression of Dor in car1 mutant aggravates this defect, implicating Car in the removal of Dor from endosomes. This suggests that, in addition to an independent role in fusion with tubular lysosomes, the Sec1p homologue, Car, regulates Dor function. PMID- 12743108 TI - Role of Drosophila Rab5 during endosomal trafficking at the synapse and evoked neurotransmitter release. AB - During constitutive endocytosis, internalized membrane traffics through endosomal compartments. At synapses, endocytosis of vesicular membrane is temporally coupled to action potential-induced exocytosis of synaptic vesicles. Endocytosed membrane may immediately be reused for a new round of neurotransmitter release without trafficking through an endosomal compartment. Using GFP-tagged endosomal markers, we monitored an endosomal compartment in Drosophila neuromuscular synapses. We showed that in conditions in which the synaptic vesicles pool is depleted, the endosome is also drastically reduced and only recovers from membrane derived by dynamin-mediated endocytosis. This suggests that membrane exchange takes place between the vesicle pool and the synaptic endosome. We demonstrate that the small GTPase Rab5 is required for endosome integrity in the presynaptic terminal. Impaired Rab5 function affects endo- and exocytosis rates and decreases the evoked neurotransmitter release probability. Conversely, Rab5 overexpression increases the release efficacy. Therefore, the Rab5-dependent trafficking pathway plays an important role for synaptic performance. PMID- 12743109 TI - Neural cell adhesion molecule (NCAM) association with PKCbeta2 via betaI spectrin is implicated in NCAM-mediated neurite outgrowth. AB - In hippocampal neurons and transfected CHO cells, neural cell adhesion molecule (NCAM) 120, NCAM140, and NCAM180 form Triton X-100-insoluble complexes with betaI spectrin. Heteromeric spectrin (alphaIbetaI) binds to the intracellular domain of NCAM180, and isolated spectrin subunits bind to both NCAM180 and NCAM140, as does the betaI spectrin fragment encompassing second and third spectrin repeats (betaI2-3). In NCAM120-transfected cells, betaI spectrin is detectable predominantly in lipid rafts. Treatment of cells with methyl-beta-cyclodextrin disrupts the NCAM120-spectrin complex, implicating lipid rafts as a platform linking NCAM120 and spectrin. NCAM140/NCAM180-betaI spectrin complexes do not depend on raft integrity and are located both in rafts and raft-free membrane domains. PKCbeta2 forms detergent-insoluble complexes with NCAM140/NCAM180 and spectrin. Activation of NCAM enhances the formation of NCAM140/NCAM180-spectrin PKCbeta2 complexes and results in their redistribution to lipid rafts. The complex is disrupted by the expression of dominant-negative betaI2-3, which impairs binding of spectrin to NCAM, implicating spectrin as the bridge between PKCbeta2 and NCAM140 or NCAM180. Redistribution of PKCbeta2 to NCAM-spectrin complexes is also blocked by a specific fibroblast growth factor receptor inhibitor. Furthermore, transfection with betaI2-3 inhibits NCAM-induced neurite outgrowth, showing that formation of the NCAM-spectrin-PKCbeta2 complex is necessary for NCAM-mediated neurite outgrowth. PMID- 12743110 TI - PMN transendothelial migration decreases nuclear NFkappaB in IL-1beta-activated endothelial cells: role of PECAM-1. AB - During the systemic inflammatory response, circulating cytokines interact with the vascular endothelium, resulting in activation and nuclear accumulation of the nuclear transcription factor, nuclear factor kappa B (NFkappaB). In turn, NFkappaB transactivates relevant proinflammatory genes, resulting in an amplification of the inflammatory response. Because this scenario is potentially detrimental to the host, mechanisms exist to limit this amplification. Using an in vitro system that mimics the vascular-interstitial interface during inflammation (cell culture inserts), we provide evidence for the existence of a novel negative feedback mechanism on NFkappaB activity. We show that the interleukin 1beta-induced accumulation of nuclear NFkappaB in human umbilical vein endothelial cell monolayers is dramatically reduced when polymorphonuclear leukocytes (PMN) are allowed to migrate across these monolayers. This effect does not appear to be due to PMN-derived elastase or nitric oxide. Fixed PMN (adhere but do not migrate) did not affect nuclear NFkappaB. Furthermore, cross-linking of platelet-endothelial cell adhesion molecule-1 (PECAM-1), but not intercellular adhesion molecule-1, reduces human umbilical vein endothelial cell nuclear NFkappaB induced by interleukin 1beta. Finally, interaction of PMN with PECAM-1 deficient endothelial cells does not reduce nuclear NFkappaB. These observations indicate that engagement of PECAM-1 by emigrating PMN is a pivotal event in this negative feedback on NFkappaB activity. PMID- 12743112 TI - Mapping the Gbetagamma-binding sites in GIRK1 and GIRK2 subunits of the G protein activated K+ channel. AB - G protein-activated K+ channels (Kir3 or GIRK) are activated by direct binding of Gbetagamma. The binding sites of Gbetagamma in the ubiquitous GIRK1 (Kir3.1) subunit have not been unequivocally charted, and in the neuronal GIRK2 (Kir3.2) subunit the binding of Gbetagamma has not been studied. We verified and extended the map of Gbetagamma-binding sites in GIRK1 by using two approaches: direct binding of Gbetagamma to fragments of GIRK subunits (pull down), and competition of these fragments with the Galphai1 subunit for binding to Gbetagamma. We also mapped the Gbetagamma-binding sites in GIRK2. In both subunits, the N terminus binds Gbetagamma. In the C terminus, the Gbetagamma-binding sites in the two subunits are not identical; GIRK1, but not GIRK2, has a previously unrecognized Gbetagamma-interacting segments in the first half of the C terminus. The main C terminal Gbetagamma-binding segment found in both subunits is located approximately between amino acids 320 and 409 (by GIRK1 count). Mutation of C terminal leucines 262 or 333 in GIRK1, recognized previously as crucial for Gbetagamma regulation of the channel, and of the corresponding leucines 273 and 344 in GIRK2 dramatically altered the properties of K+ currents via GIRK1/GIRK2 channels expressed in Xenopus oocytes but did not appreciably reduce the binding of Gbetagamma to the corresponding fusion proteins, indicating that these residues are mainly important for the regulation of Gbetagamma-induced changes in channel gating rather than Gbetagamma binding. PMID- 12743113 TI - Reversible inhibition of cytochrome c oxidase by peroxynitrite proceeds through ascorbate-dependent generation of nitric oxide. AB - Reversible inhibition of cytochrome c oxidase (CcOX) by nitric oxide (NO*) has potential physiological roles in the regulation of mitochondrial respiration, redox signaling, and apoptosis. However peroxynitrite (ONOO-), an oxidant formed from the reaction of NO* and superoxide, appears mostly detrimental to cell function. This occurs both through direct oxidant reactions and by decreasing the availability of NO* for interacting with CcOX. When isolated CcOX respires with ascorbate as a reducing substrate, the conversion of ONOO- to NO* is observed. It is not known whether this can be ascribed to a direct interaction of the enzyme with ONOO-. In this investigation, the role of ascorbate in this system was examined using polarographic methods to measure NO* production and CcOX activity simultaneously in both the purified enzyme and isolated mitochondria. It was found that ascorbate alone accounts for >90% of the NO* yield from ONOO- in the presence or absence of purified CcOX in turnover. The yield of NO was CcOX independent but was dependent on ascorbate and ONOO- concentrations and was not affected by metal chelators. Consistent with this, the interaction of ONOO- with CcOX in respiring isolated mitochondria only yielded NO* when ascorbate was also present in the incubation. These observations are discussed in the context of ONOO-/ascorbate reactivity and the interaction of CcOX with reactive nitrogen species. PMID- 12743111 TI - Size-selective loosening of the blood-brain barrier in claudin-5-deficient mice. AB - Tight junctions are well-developed between adjacent endothelial cells of blood vessels in the central nervous system, and play a central role in establishing the blood-brain barrier (BBB). Claudin-5 is a major cell adhesion molecule of tight junctions in brain endothelial cells. To examine its possible involvement in the BBB, claudin-5-deficient mice were generated. In the brains of these mice, the development and morphology of blood vessels were not altered, showing no bleeding or edema. However, tracer experiments and magnetic resonance imaging revealed that in these mice, the BBB against small molecules (<800 D), but not larger molecules, was selectively affected. This unexpected finding (i.e., the size-selective loosening of the BBB) not only provides new insight into the basic molecular physiology of BBB but also opens a new way to deliver potential drugs across the BBB into the central nervous system. PMID- 12743114 TI - Tissue transglutaminase directly regulates adenylyl cyclase resulting in enhanced cAMP-response element-binding protein (CREB) activation. AB - Tissue transglutaminase (tTG) is present in the human nervous system and is predominantly localized to neurons. Treatment of human neuroblastoma SH-SY5Y cells with retinoic acid results in increased tTG expression, which is both necessary and sufficient for differentiation. The goal of the present study was to determine whether tTG modulates the activation of the cyclic AMP-response element (CRE)-binding protein, CREB, an event that likely plays a central role in the differentiation of SH-SY5Y cells. SH-SY5Y cells stably transfected with active wild type tTG, tTG without transamidating activity (C277S), an antisense tTG construct that depleted the endogenous levels of tTG, or vector only were used for the study. Treatment with forskolin, an adenylyl cyclase activator, increased that activation-associated phosphorylation of CREB, which was prolonged by tTG overexpression. CRE-reporter gene activity was also significantly elevated in the tTG cells compared with the other cells. The enhancement of CREB phosphorylation/activation in the tTG cells is likely due to the fact that tTG significantly potentiates cAMP production, and our findings indicate that tTG enhances adenylyl cyclase activity by modulating the conformation state of adenylyl cyclase. This is the first study to provide evidence of the mechanism by which tTG may contribute to neuronal differentiation. PMID- 12743115 TI - The obligate predatory Bdellovibrio bacteriovorus possesses a neutral lipid A containing alpha-D-Mannoses that replace phosphate residues: similarities and differences between the lipid As and the lipopolysaccharides of the wild type strain B. bacteriovorus HD100 and its host-independent derivative HI100. AB - Bdellovibrio bacteriovorus are predatory bacteria that penetrate Gram-negative bacteria and grow intraperiplasmically at the expense of the prey. It was suggested that B. bacteriovorus partially degrade and reutilize lipopolysaccharide (LPS) of the host, thus synthesizing an outer membrane containing structural elements of the prey. According to this hypothesis a host independent mutant should possess a chemically different LPS. Therefore, the lipopolysaccharides of B. bacteriovorus HD100 and its host-independent derivative B. bacteriovorus HI100 were isolated and characterized by SDS-polyacrylamide gel electrophoresis, immunoblotting, and mass spectrometry. LPS of both strains were identified as smooth-form LPS with different repeating units. The lipid As were isolated after mild acid hydrolysis and their structures were determined by chemical analysis, by mass spectrometric methods, and by NMR spectroscopy. Both lipid As were characterized by an unusual chemical structure, consisting of a beta-(1-->6)-linked 2,3-diamino-2,3-dideoxy-d-glucopyranose disaccharide carrying six fatty acids that were all hydroxylated. Instead of phosphate groups substituting position O-1 of the reducing and O-4' of the nonreducing end alpha-d mannopyranose residues were found in these lipid As. Thus, they represent the first lipid As completely missing negatively charged groups. A reduced endotoxic activity as determined by cytokine induction from human macrophages was shown for this novel structure. Only minor differences with respect to fatty acids were detected between the lipid As of the host-dependent wild type strain HD100 and for its host-independent derivative HI100. From the results of the detailed analysis it can be concluded that the wild type strain HD100 synthesizes an innate LPS. PMID- 12743116 TI - Crystal structure of the intrinsically flexible addiction antidote MazE. AB - A specific camel VHH (variable domain of dromedary heavy chain antibody) fragment was used to crystallize the intrinsically flexible addiction antidote MazE. Only 45% of the polypeptide chain is found ordered in the crystal. The MazE monomer consisting of two beta-hairpins connected by a short alpha-helix has no hydrophobic core on its own and represents only one half of a typical protein domain. A complete domain structure is formed by the association of two chains, creating a hydrophobic core between two four-stranded beta-sheets. This hydrophobic core consists exclusively of short aliphatic residues. The folded part of MazE contains a novel DNA binding motif. A model for DNA binding that is consistent with the available biochemical data is presented. PMID- 12743117 TI - Glycosylphosphatidylinositol-anchored ceruloplasmin is required for iron efflux from cells in the central nervous system. AB - Ceruloplasmin (Cp) is a ferroxidase that converts highly toxic ferrous iron to its non-toxic ferric form. A glycosylphosphatidylinositol (GPI)-anchored form of this enzyme is expressed by astrocytes in the mammalian central nervous system, whereas the secreted form is expressed by the liver and found in serum. Lack of this enzyme results in iron accumulation in the brain and neurodegeneration. Herein, we show using astrocytes purified from the central nervous system of Cp null mice that GPI-Cp is essential for iron efflux and not involved in regulating iron influx. We also show that GPI-Cp colocalizes on the astrocyte cell surface with the divalent metal transporter IREG1 and is physically associated with IREG1. In addition, IREG1 alone is unable to efflux iron from astrocytes in the absence of GPI-Cp or secreted Cp. We also provide evidence that the divalent metal influx transporter DMT1 is expressed by astrocytes and is likely to mediate iron influx into these glial cells. The coordinated actions of GPI-Cp and IREG1 may be required for iron efflux from neural cells, and disruption of this balance could lead to iron accumulation in the central nervous system and neurodegeneration. PMID- 12743119 TI - Binding site selection for the plant MADS domain protein AGL15: an in vitro and in vivo study. AB - AGL15 (for AGAMOUS-like 15) is currently the only reported member of the plant MADS domain family of transcriptional regulators that preferentially accumulates during embryo development. Additionally, AGL15 is one of the more divergent members of the MADS domain family, including within the DNA-binding domain. Previous studies have shown that MADS domain proteins bind to DNA sequences with an overall consensus of CC(A/T)6GG (called a CArG motif). Nonetheless, different MADS domain proteins exhibit similar yet distinct binding site preferences that may be critical for differential gene regulation. To determine the consensus sequence preferentially bound by AGL15 in vitro, PCR-assisted binding site selection assays were performed. AGL15 was observed to prefer a CArG motif with a longer A/T-rich core and is to date the only plant MADS domain protein having such a preference. Next, the Arabidopsis genome data base was searched for genes containing AGL15 binding sites as candidates for direct regulation by AGL15. One gene, DTA4 (for Downstream Target of AGL15-4), was identified by this method, and then confirmed as a direct target of AGL15 in vivo. PMID- 12743118 TI - The influence of conserved aromatic residues in 3-hydroxy-3-methylglutaryl-CoA synthase. AB - In order to evaluate the potential contribution of conserved aromatic residues to the hydrophobic active site of 3-hydroxy-3-methylglutaryl-CoA synthase, site directed mutagenesis was employed to produce Y130L, Y163L, F204L, Y225L, Y346L, and Y376L proteins. Each mutant protein was expressed at levels comparable with wild-type enzyme and was isolated in highly purified form. Initial kinetic characterization indicated that F204L exhibits a substantial (>300-fold) decrease in catalytic rate (kcat). Upon modification with the mechanism-based inhibitor, 3 chloropropionyl-CoA, or in formation of a stable binary complex with acetoacetyl CoA, F204L exhibits binding stoichiometries comparable with wild-type enzyme, suggesting substantial retention of active site integrity. Y130L and Y376L exhibit inflated values (80- and 40-fold, respectively) for the Km for acetyl-CoA in the acetyl-CoA hydrolysis partial reaction; these mutants also exhibit an order of magnitude decrease in kcat. Formation of the acetyl-S-enzyme reaction intermediate by Y130L, F204L, and Y376L proceeds slowly in comparison with wild type enzyme. However, solvent exchange into the thioester carbonyl oxygen of these acetyl-S-enzyme intermediates is not slow in comparison with previous observations for D159A and D203A mutants, which also exhibit slow acetyl-S-enzyme formation. The magnitude of the differential isotope shift upon exchange of H218O into [13C]acetyl-S-enzyme suggests a polarization of the thioester carbonyl and a reduction in bond order. Such an effect may substantially contribute to the upfield 13C NMR shift observed for [13C]acetyl-S-enzyme. The influence on acetyl S-enzyme formation, as well as observed kcat (F204L) and Km (Y130L; Y376L) effects, implicate these invariant residues as part of the catalytic site. Substitution of phenylalanine (Y130F, Y376F) instead of leucine at residues 130 and 376 diminishes the effects on catalytic rate and substrate affinity observed for Y130L and Y376L, underscoring the influence of aromatic side chains near the active site. PMID- 12743120 TI - Parathyroid hormone-induced E4BP4/NFIL3 down-regulates transcription in osteoblasts. AB - Parathyroid hormone (PTH), a major regulator of bone metabolism, activates the PTHR1 receptor on the osteoblast plasma membrane to initiate signaling and induce transcription of primary response genes. Subsequently, primary genes with transcriptional activity regulate expression of downstream PTH targets. We have identified the adenovirus E4 promoter-binding protein/nuclear factor regulated by IL-3 (E4bp4) as a PTH-induced primary gene in osteoblasts. E4BP4 is a basic leucine zipper (bZIP) transcription factor that represses or activates transcription in non-osteoblastic cells. We report here that PTH rapidly and transiently induced E4bp4 mRNA in osteoblastic cells and that this induction did not require protein synthesis. PTH also induced E4BP4 protein synthesis and E4BP4 binding to a consensus but not to a mutant E4BP4 response element (EBPRE). E4BP4 overexpression inhibited an EBPRE-containing promoter-reporter construct, whereas PTH treatment attenuated activity of the same construct in primary mouse osteoblasts. Finally, E4BP4 overexpression inhibited PTH-induced activity of a cyclooxygenase-2 promoter-reporter construct. Our data suggest a role for E4BP4 in attenuation of PTH target gene transcription in osteoblasts. PMID- 12743121 TI - Transforming growth factor-beta1 produced by ovarian cancer cell line HRA stimulates attachment and invasion through an up-regulation of plasminogen activator inhibitor type-1 in human peritoneal mesothelial cells. AB - The processes of ovarian cancer dissemination are characterized by altered local proteolysis, cellular proliferation, cell attachment, and invasion, suggesting that the urokinase-type plasminogen activator (uPA) and its specific inhibitor (plasminogen activator inhibitor type-1 (PAI-1)) could be involved in the pathogenesis of peritoneal dissemination. We showed previously that expression of uPA and PAI-1 in the human ovarian cancer cell line HRA can be down-regulated by exogenous bikunin (bik), a Kunitz-type protease inhibitor, via suppression of transforming growth factor-beta1 (TGF-beta1) up-regulation and that overexpression of the bik gene can specifically suppress the in vivo growth and peritoneal dissemination of HRA cells in an animal model. We hypothesize that the plasminogen activator system in mesothelial cells can be modulated by HRA cells. To test this hypothesis, we used complementary techniques in mesothelial cells to determine whether uPA and PAI-1 expression are altered by exposure to culture media conditioned by HRA cells. Here we show the following: 1) that expression of PAI-1, but not uPA, was markedly induced by culture media conditioned by wild type HRA cells but not by bik transfected clones; 2) that by antibody neutralization the effect appeared to be mediated by HRA cell-derived TGF-beta1; 3) that exogenous TGF-beta1 specifically enhanced PAI-1 up-regulation at the mRNA and protein level in mesothelial cells in a time- and concentration-dependent manner, mainly through MAPK-dependent activation mechanism; and 4) that mesothelial cell-derived PAI-1 may promote tumor invasion possibly by enhancing cell-cell interaction. This represents a novel pathway by which tumor cells can regulate the plasminogen activator system-dependent cellular responses in mesothelial cells that may contribute to formation of peritoneal dissemination of ovarian cancer. PMID- 12743122 TI - Thermotoga maritima 3-deoxy-D-arabino-heptulosonate 7-phosphate (DAHP) synthase: the ancestral eubacterial DAHP synthase? AB - The gene encoding the 3-deoxy-d-arabino-heptulosonate 7-phosphate (DAHP) synthase from the thermophilic microorganism Thermotoga maritima was cloned, and the enzyme was overexpressed in Escherichia coli. The purified DAHP synthase displays a homotetrameric structure and exhibits maximal activity at 90 degrees C. The enzyme is extremely thermostable, with 50% of its initial activity retained after incubation for approximately 5 h at 80 degrees C, 21 h at 70 degrees C, and 86 h at 60 degrees C. The enzyme appears to follow Michaelis-Menten kinetics with Km for phosphoenolpyruvate = 9.5-13 microm, Km for d-erythrose 4-phosphate = 57.3 350.1 microm, and kcat = 2.3-7.6 s-1 between 50 degrees C and 70 degrees C. Metal analysis indicates that DAHP synthase as isolated contains Zn2+, and the enzyme is inactivated by treatment with EDTA. The apo-enzyme is partially reactivated by a variety of divalent metals including Zn2+, Cd2+, Mn2+, Cu2+, Co2+, and Ni2+. These observations suggest that T. maritima DAHP synthase is a metalloenzyme. The activity of T. maritima DAHP synthase is inhibited by two of the three aromatic amino acids (l-Phe and l-Tyr) formed in the Shikimate pathway. This report is the first description of a thermophilic eubacterial DAHP synthase. PMID- 12743123 TI - Ybp1 is required for the hydrogen peroxide-induced oxidation of the Yap1 transcription factor. AB - We describe the characterization of Ybp1, a novel protein, in Saccharomyces cerevisiae, that is required for the oxidative stress response to peroxides. Ybp1 is required for H2O2-induced expression of the antioxidant encoding gene TRX2. Our data indicate that the effects of Ybp1 are mediated through the Yap1 transcription factor. Indeed, Ybp1 forms a stress-induced complex with Yap1 in vivo and stimulates the nuclear accumulation of Yap1 in response to H2O2 but not in response to the thiol-oxidizing agent diamide. The H2O2-induced nuclear accumulation of Yap1 is regulated by the oxidation of specific cysteine residues and is dependent on the thiol peroxidase Gpx3. Our data suggest that Ybp1 is required for the H2O2-induced oxidation of Yap1 and acts in the same pathway as Gpx3. Consequently, Ybp1 represents a novel class of stress regulator of Yap1. These data have important implications for the regulation of protein oxidation and stress responses in eukaryotes. PMID- 12743125 TI - Effects of overexpression of copper-zinc and manganese superoxide dismutases, catalase, and thioredoxin reductase genes on longevity in Drosophila melanogaster. AB - The overexpression of antioxidative enzymes such as CuZn-superoxide dismutase (SOD), Mn-SOD, and catalase has previously been reported to extend life span in transgenic flies (Drosophila melanogaster). The purpose of this study was to determine whether life-extending effects persist if the recipient control strains of flies are relatively long-lived. Accordingly, the life spans of large numbers of replicate control and overexpressor lines were determined in two long-lived genetic backgrounds involving a combined total of >90,000 flies. Significant increases in the activities of both CuZn-SOD and catalase had no beneficial effect on survivorship in relatively long-lived y w mutant flies and were associated with slightly decreased life spans in wild type flies of the Oregon-R strain. The introduction of additional transgenes encoding Mn-SOD or thioredoxin reductase in the same genetic background also failed to cause life span extension. In conjunction with data from earlier studies, the results show that increasing the activities of these major antioxidative enzymes above wild type levels does not decrease the rate of aging in long-lived strains of Drosophila, although there may be some effect in relatively short-lived strains. PMID- 12743124 TI - Zinc-induced PTEN protein degradation through the proteasome pathway in human airway epithelial cells. AB - The tumor suppressor PTEN is a putative negative regulator of the phosphatidylinositol 3-kinase/Akt pathway. Exposure to Zn2+ ions induces Akt activation, suggesting that PTEN may be modulated in this process. Therefore, the effects of Zn2+ on PTEN were studied in human airway epithelial cells and rat lungs. Treatment with Zn2+ resulted in a significant reduction in levels of PTEN protein in a dose- and time-dependent fashion in a human airway epithelial cell line. This effect of Zn2+was also observed in normal human airway epithelial cells in primary culture and in rat airway epithelium in vivo. Concomitantly, levels of PTEN mRNA were also significantly reduced by Zn2+ exposure. PTEN phosphatase activity evaluated by measuring Akt phosphorylation decreased after Zn2+ treatment. Pretreatment of the cells with a proteasome inhibitor significantly blocked zinc-induced reduction of PTEN protein as well as the increase in Akt phosphorylation, implicating the involvement of proteasome mediated PTEN degradation. Further study revealed that Zn2+-induced ubiquitination of PTEN protein may mediate this process. A phosphatidylinositol 3 kinase inhibitor blocked PTEN degradation induced by Zn2+, suggesting that phosphatidylinositol 3-kinase may participate in the regulation of PTEN. However, both the proteasome inhibitor and phosphatidylinositol 3-kinase inhibitor failed to prevent significant down-regulation of PTEN mRNA expression in response to Zn2+. In summary, exposure to Zn2+ ions causes PTEN degradation and loss of function, which is mediated by an ubiquitin-associated proteolytic process in the airway epithelium. PMID- 12743126 TI - Shear-induced cyclooxygenase-2 via a JNK2/c-Jun-dependent pathway regulates prostaglandin receptor expression in chondrocytic cells. AB - Using cDNA microarrays coupled with bioinformatics tools, we elucidated a signaling cascade regulating cyclooxygenase-2 (COX-2), a pivotal pro-inflammatory enzyme expressed in rheumatic and osteoarthritic, but not normal, cartilage. Exposure of T/C-28a2 chondrocytic cells to fluid shear results in co-regulation of c-Jun N-terminal kinase2 (JNK2), c-Jun, and COX-2 as well as concomitant downstream expression of prostaglandin receptors EP2 and EP3a1. JNK2 transcript inhibition abrogated shear-induced COX-2, EP2, and EP3a1 mRNA up-regulation as well as c-Jun phosphorylation. Functional knock-out experiments using an antisense c-Jun oligonucleotide revealed the abolition of shear-induced COX-2, EP2, and EP3a1, but not JNK2, transcripts. Moreover, inhibition of COX-2 activity eliminated mRNA upregulation of EP2 and EP3a1 induced by shear. Hence, a biochemical pathway exists wherein fluid shear activates COX-2, via a JNK2/c-Jun dependent pathway, which in turn elicits downstream EP2 and EP3a1 mRNA synthesis. PMID- 12743128 TI - Current status of taxane and platinum-based chemotherapy in ovarian cancer. AB - Before 1993, the standard of care for the chemotherapeutic management of advanced ovarian cancer was cisplatin or carboplatin combined with a classic alkylating agent (typically cyclophosphamide). Studies in the 1990s have changed this standard to one of the platinum-containing agents combined with a taxane, paclitaxel, or docetaxel. This article reviews the pertinent studies regarding paclitaxel combined with cisplatin or carboplatin, discusses the remaining controversies surrounding how best to combine these agents, and provides opinions regarding the discordant outcomes noted in studies of the paclitaxel-platinum doublet. A separate article discusses the docetaxel-platinum doublet and how that might be considered an appropriate option for first-line therapy in patients with advanced, newly diagnosed ovarian cancer. PMID- 12743127 TI - Progress in the management of gynecologic cancer: consensus summary statement. PMID- 12743129 TI - Role of docetaxel in the treatment of newly diagnosed advanced ovarian cancer. AB - Docetaxel is currently licensed for use in a variety of malignancies, including breast cancer and lung cancer, and is the preferred taxane in breast cancer treatment. In ovarian cancer, the taxane of choice has historically been paclitaxel; however, there is now substantial evidence that docetaxel also may be the preferred taxane in this disease. Docetaxel has many preclinical advantages over paclitaxel and has been shown to be effective in both platinum- and paclitaxel-resistant disease. Phase I and II studies have shown docetaxel plus carboplatin to be feasible, and the combination is associated with a tolerable adverse-effect profile. The Scottish Randomized Trial in Ovarian Cancer (SCOTROC) trial randomly assigned 1,077 patients with International Federation of Gynecology and Obstetrics stage Ic to IV disease to six cycles of docetaxel plus carboplatin (DC) or paclitaxel plus carboplatin (PC) as primary chemotherapy. Progression-free survival is not statistically different, and to date, no differences are apparent in overall survival. Toxicity differences were evident. There was more myelosuppression with DC but no additional mortality. More neuropathy was present with PC, with more patients stopping paclitaxel because of this toxicity during chemotherapy. Quality-of-life analyses highlighted important differences, all favoring the DC treatment arm. Additional SCOTROC studies using docetaxel are ongoing. These data indicate that docetaxel and carboplatin represent a reasonable first-line option for patients with newly diagnosed epithelial ovarian cancer. PMID- 12743130 TI - Role of intraperitoneal chemotherapy in the front-line setting. AB - During the last several decades, clinical investigators have confirmed the safety and pharmacokinetic advantage associated with the intraperitoneal delivery of a number of antineoplastic agents with known activity in ovarian cancer. Phase II experience has revealed that objective responses, including surgically documented complete remissions, can occur when regional drug delivery is used as second-line treatment of this malignancy. Three randomized trials have shown that when cisplatin-based intraperitoneal treatment is used in the front-line setting, it results in superior progression-free and overall survival compared with intravenous cisplatin. Despite these facts, an ultimate role for regional drug delivery as initial treatment of ovarian cancer will require additional trials to define how to optimize the use of this unique therapeutic strategy. PMID- 12743131 TI - Biologic and immunologic therapies for ovarian cancer. AB - Biologic therapy of ovarian cancer has been conducted using nonspecific biologic response modifiers, cytokines, monoclonal antibodies (MAbs), vaccines, and gene therapy. Antibodies directed toward her2/neu have also been studied. Phase I and II gene therapy trials using adenoviral vectors containing a wild-type or modified p53 have shown that the treatment is well tolerated. Phase II and III trials are ongoing with MAbs directed against CA-125 (MAb B43.13) and an antibody directed against HMFG1 (anti-HMFG1-yttrium-90-labeled antibody). The trials have shown that these agents are well tolerated and that immunologic responses occur, although the ultimate clinical value of these agents remains to be determined. Prolonged survival after MAb B43.13 treatment has been correlated with changes in several immune parameters, including human antimurine antibody, Ab2, anti-CA-125 antibody development, and induced T-cell immunity. Clinical trials using a MAb directed toward the encoded products of her2/neu have shown minimal activity against ovarian cancer in a phase I and II trial conducted by the Gynecologic Oncology Group. Cytokine therapies have been administered systemically and intraperitoneally. Intracavitary interferon alfa, interferon gamma, and interleukin-2 alone or in combination with cytotoxic therapy in phase I and II trials demonstrated intraperitoneal lymphoid cell stimulation and produced antitumor responses. A randomized trial of chemotherapy with or without interferon gamma in primary treatment produced a response and a progression-free survival advantage in the arm that incorporated the interferon gamma, without a statistically significant benefit in overall survival. A phase III study of interferon gamma in combination with first-line chemotherapy is currently ongoing. PMID- 12743133 TI - Use of CA-125 to assess response to new agents in ovarian cancer trials. AB - The purpose of this article is to demonstrate how CA-125 could be used in clinical trials to ascertain the efficacy of new drugs for ovarian cancer. Studies that have investigated the use of CA-125 in assessing response and progression of ovarian cancer are reviewed. A precise CA-125 response definition that requires either a 50% or 75% decrease in CA-125 levels has been shown in trials of both initial chemotherapy and in phase II trials to predict accurately the response in comparison with standard response criteria. A simpler response definition that is based on just a 50% decrease in CA-125 levels has been proposed by the Gynaecological Cancer Intergroup (GCIG) but requires further validation. Definitions for progression have also been proposed by the GCIG on the basis of a confirmed doubling of CA-125 levels from either the upper limit of normal or the nadir CA-125 level. These CA-125 definitions for progression falsely predict progression in fewer than 2% of patients and can be used to define the date of progression. Precise definitions for response and progression according to CA-125 should be incorporated into ovarian cancer clinical trial protocols. These definitions already have been shown to be valuable in assessing efficacy of new agents but require further prospective evaluation. PMID- 12743132 TI - Challenges of developing therapeutics that target signal transduction in patients with gynecologic and other malignancies. AB - A greater understanding of cancer biology and major advances in biotechnology have resulted in the identification of a plethora of rationally designed, target based anticancer therapeutics, particularly those that inhibit malignant-cell signal transduction, ushering in new therapeutic opportunities and extraordinary developmental challenges. Because these agents seem to principally target malignant cells, it is expected that they will produce less toxicity at clinically effective doses than nonspecific cytotoxic agents. The innate complexity of signaling networks, which have redundant relay systems that confer robustness, adaptability, and signaling diversity, also decreases the probability that any single therapeutic manipulation against any specific signaling element will be highly successful when used alone, particularly in patients with solid malignancies that have multiple relevant signaling aberrations. In addition, because the predominant therapeutic effect of inhibitors of signal transduction processes in preclinical studies is a decreased rate of tumor growth, it is anticipated that the predominant therapeutic outcome in the clinic will be similar; however, this end point is not readily detectable or quantifiable using traditional clinical evaluation methods. Furthermore, the results of preclinical and early clinical studies indicate that dose-toxicity relationships are not likely to be as steep as with nonspecific cytotoxic agents. Therefore, both regulatory and clinical practice end points, such as time to disease progression, disease-related symptoms, and quality of life, which are generally considered secondary for cytotoxic agents, may evolve into primary end points. The cumulative results of developmental evaluations to date indicate that the development, evaluation, and general clinical use of rationally designed, target based anticancer therapeutics will require a radical departure from traditional paradigms to exploit the full potential of these new therapies. PMID- 12743134 TI - Ultrasound screening for the early detection of ovarian cancer. AB - Ovarian cancer screening in the general population has been performed using ultrasound examination of the female pelvis and serum tumor marker determinations. Ultrasound examinations, particularly transvaginal sonography (TVS), have been advocated as potentially useful modalities. Investigators from the University of Kentucky (Lexington, KY) and Hirosaki University (Hirosaki, Japan) have recently published results from ultrasound-based ovarian cancer screening studies. The Kentucky trial screened 14,469 women using TVS on an annual basis. One hundred eighty women underwent surgery, and 17 ovarian cancers were detected, 11 of which were invasive epithelial lesions. The Hirosaki trial reported the results of an ultrasound-based screening study among 51,550 women who were first-time participants. Three hundred twenty-four women underwent surgery, and 22 ovarian cancers were detected. In each of these trials, the positive predictive value of gray-scale sonography was low. Morphologic tumor indexing and Doppler examinations have both been proposed as potential second line studies, which could increase the positive predictive value of gray-scale ultrasound. A review of these techniques is presented. At present, ovarian cancer screening in the general population using ultrasound examinations is an experimental technique. Further studies are needed to determine whether second line testing can improve the positive predictive value of gray-scale sonography such that asymptomatic women do not undergo unnecessary surgery for benign masses. PMID- 12743135 TI - Status of tumor markers in ovarian cancer screening. AB - One of the most promising approaches to management of ovarian cancer is early detection. Stage I ovarian cancer can be cured with currently available therapy in more than 90% of patients. However, fewer than 25% of ovarian cancers are currently detected in stage I. Detection of a greater fraction of cancers at an early stage might improve clinical outcome. Given a prevalence of one patient with ovarian cancer among 2,500 asymptomatic postmenopausal women in the general population, a successful screening strategy must have a sensitivity of more than 75% and a specificity of more than 99.6% to achieve a positive predictive value of 10%. Approaches to screening include transvaginal sonography, serum markers, and two-stage strategies that use alterations in serum markers to prompt sonographic examination. Among the serum markers, CA-125 has been studied most extensively. Isolated values of CA-125 lack adequate sensitivity or specificity, but when monitored over time, serial CA-125 values can achieve a specificity of 99.6%. However, sensitivity is limited and CA-125 may only be expressed by 80% of early-stage cancers. Multiple markers may exhibit greater specificity when studied over time. To combine multiple markers, more sophisticated mathematical analysis will be required. At present, screening women at conventional risk should be restricted to clinical trials. In the future, however, screening for ovarian cancer may reduce the morbidity and mortality of this disease. PMID- 12743137 TI - Combined-modality therapy of locally advanced cervical cancer. AB - Six randomized trials of chemoradiotherapy in cervical cancer have demonstrated improvement with cisplatin-based chemoradiotherapy compared with radiation alone or radiation with hydroxyurea. Only one randomized trial (Gynecologic Oncology Group [GOG] 120) compared different cisplatin-based regimens and demonstrated significantly more hematologic and gastrointestinal toxic effects with the combination of cisplatin, fluorouracil, and hydroxyurea than with weekly cisplatin alone. The use of fluorouracil and mitomycin as radiation sensitizers for cervical cancer is based on their activity in chemoradiotherapy for anal carcinoma. However, a three-fold increase in serious late bowel toxicity was seen in patients treated with fluorouracil and mitomycin compared with fluorouracil alone when combined with radiation therapy for cervical cancer. Another randomized trial demonstrated improvement in survival with fluorouracil infusion compared with radiation alone only in early-stage disease. However, the most recently completed randomized trial by the GOG, which compared continuous fluorouracil infusion with weekly cisplatin, was closed when interim analysis demonstrated no potential superiority to the fluorouracil arm. Uncontrolled studies have evaluated the use of carboplatin on several different schedules, but no controlled trials have yet been performed. In metastatic and recurrent cervical cancer, a number of newer agents, including paclitaxel, gemcitabine, topotecan, and vinorelbine, have demonstrated modest single-agent activity and even greater activity in combination with cisplatin. An expanding series of trials will define the role of these new agents combined with cisplatin and radiation therapy for locally advanced cervical cancer. The potential role of biologic agents in combination with chemoradiotherapy also needs to be examined in locally advanced cervical cancer. PMID- 12743136 TI - Calculation of the risk of ovarian cancer from serial CA-125 values for preclinical detection in postmenopausal women. AB - PURPOSE: Previous studies of CA-125 levels from screening trials for ovarian cancer have indicated that serial CA-125 levels may identify cases better than a fixed CA-125 cutoff. We conducted a study to assess the screening performance of the risk of ovarian cancer calculation based on serial CA-125 levels from prospectively collected serum samples compared with a fixed CA-125 cutoff. PATIENTS AND METHODS: The calculation was applied to data from a prospective trial of screening for ovarian cancer involving 22,000 postmenopausal women older than 45 years. The analysis was performed using 33,621 CA-125 results from 9,233 women for whom two or more serial samples were available. All serum samples from the patients with ovarian cancer were obtained before clinical detection. Sensitivity and specificity levels for preclinical detection of index cancers were calculated for various cutoffs for the risk and a single CA-125 measurement, and receiver operator curves were constructed. RESULTS: The risk calculation significantly improved the area under the curve from 84% to 93% compared with a fixed cutoff for CA-125 (P =.01). For a target specificity of 98%, the risk achieved a sensitivity of 86% for preclinical detection of ovarian cancer, whereas CA-125 achieved a sensitivity of 62%. The estimates of performance are unbiased, because the risk calculation was derived independent of the data from this trial. CONCLUSION: These results provide the first evidence that preclinical detection of ovarian cancer using serial CA-125 levels interpreted with the risk calculation significantly improves screening performance compared with a fixed cutoff for CA-125. The results justify the incorporation of the risk calculation in a prospective, randomized, controlled trial. PMID- 12743138 TI - Controversies in the radiotherapeutic management of cervical cancer. AB - Radiotherapy is a critical component of treatment for many patients with cervical cancer. The role of concurrent cisplatin-based chemotherapy in the majority of cases where radiotherapy is indicated has been established. However, optimal planning and delivery of radiotherapy require attention to multiple factors beyond combined-modality chemoradiation and the continuing search for novel and effective synergistic systemic agents. Several important issues surrounding radiotherapy for cervical cancer that require clarification include the following: the effect of anemia and tumor hypoxia on outcome and their potential as therapeutic targets, the appropriate incorporation of contemporary imaging and high-technology treatment planning systems in routine clinical practice, the role of prophylactic para-aortic radiation, and the role of radioprotectors. Ongoing and newly proposed studies are expected to provide insights into these questions, which will ultimately lead to enhanced radiotherapeutic and overall care for patients with cervical cancer. PMID- 12743139 TI - Cervical cancer screening: from the Papanicolaou smear to the vaccine era. AB - In the next 20 years, cervical cancer screening will have evolved through four phases. The first was traditional screening, which has been associated with a two thirds reduction in cancer incidence and death rates in the last 50 years and currently is ending. We are entering a second phase, human papillomavirus (HPV) testing, for managing cytologic abnormalities and possibly for primary screening. A third phase, new in development, proposes the use of host biomarkers (or combinations thereof) as either surrogates of HPV infection or, potentially, indicators to assess cancer risk and concentrate available resources on a subset of women. The fourth and, likely, final phase will be screening in an era of vaccines. If HPV vaccines are successful, the pool of at-risk individuals and the prevalence of papillomaviruses that place them at risk will gradually shrink. In this climate, screening strategies that target HPVs alone (as opposed to cytologic testing) may become more economical. If so, previous strategies may become obsolete as the balance of cervical cancer prevention shifts from traditional screening to primary prevention coupled with HPV testing. PMID- 12743140 TI - A call for more with a desire for less: pelvic radiotherapy with androgen deprivation in the treatment of prostate cancer. PMID- 12743141 TI - Quality of care in prostate cancer: important to start and too important to stop here. PMID- 12743142 TI - Phase III trial comparing whole-pelvic versus prostate-only radiotherapy and neoadjuvant versus adjuvant combined androgen suppression: Radiation Therapy Oncology Group 9413. AB - PURPOSE: This trial tested the hypothesis that combined androgen suppression (CAS) and whole-pelvic (WP) radiotherapy (RT) followed by a boost to the prostate improves progression-free survival (PFS) by 10% compared with CAS and prostate only (PO) RT. This trial also tested the hypothesis that neoadjuvant and concurrent hormonal therapy (NCHT) improves PFS compared with adjuvant hormonal therapy (AHT) by 10%. MATERIALS AND METHODS: Eligibility included localized prostate cancer with an elevated prostate-specific antigen (PSA) < or = 100 ng/mL and an estimated risk of lymph node (LN) involvement of 15%. Between April 1, 1995, and June 1, 1999, 1,323 patients were accrued. Patients were randomly assigned to WP + NCHT, PO + NCHT, WP + AHT, or PO + AHT. Failure for PFS was defined as the first occurrence of local, regional, or distant disease; PSA failure; or death for any cause. RESULTS: With a median follow-up of 59.5 months, WP RT was associated with a 4-year PFS of 54% compared with 47% in patients treated with PO RT (P =.022). Patients treated with NCHT experienced a 4-year PFS of 52% versus 49% for AHT (P =.56). When comparing all four arms, there was a progression-free difference among WP RT + NCHT, PO RT + NCHT, WP RT + AHT, and PO RT + AHT (60% v 44% v 49% v 50%, respectively; P =.008). No survival advantage has yet been seen. CONCLUSION: WP RT + NCHT improves PFS compared with PO RT and NCHT or PO RT and AHT, and compared with WP RT + AHT in patients with a risk of LN involvement of 15%. PMID- 12743144 TI - Multicenter study of human immunodeficiency virus-related germ cell tumors. AB - PURPOSE: Testicular germ cell tumors (GCT) occur at increased frequency in men with human immunodeficiency virus (HIV). This multicenter study addresses the characteristics of these tumors. PATIENTS AND METHODS: Patients with HIV-related GCT were identified from six HIV treatment centers. The incidence was calculated from the center with the most complete linked oncology and HIV databases. RESULTS: Thirty-five patients with HIV-related GCT were identified. The median age at GCT diagnosis was 34 years (range, 27 to 64 years). The median CD4 cell count was 315/mm3 (range, 90 to 960/mm3) at this time. The histologic classification was seminoma in 26 patients (74%) and nonseminomatous GCT in nine patients (26%). Twenty-one patients (60%) had stage I disease and 14 patients had metastatic disease. Overall six patients relapsed, three died from GCT, and seven died from HIV disease, resulting in a 2-year overall survival rate of 81%. HIV related seminoma occurred more frequently than in the age- and sex-matched HIV negative population, with a relative risk of 5.4 (95% confidence interval, 3.35 to 8.10); however, nonseminomatous GCT did not occur more frequently, and there was no change in the incidence of GCT since the introduction of highly active antiretroviral therapy. CONCLUSION: Testicular seminoma occurs significantly more frequently in HIV-positive men than in the matched control population. Patients with HIV-related GCTs present and should be treated in a similar manner to those in the HIV-negative population. After a median follow-up of 4.6 years, 9% of the patients died from GCT. Most of the mortality relates to HIV infection. PMID- 12743143 TI - Molecular grading of urothelial cell carcinoma with fibroblast growth factor receptor 3 and MIB-1 is superior to pathologic grade for the prediction of clinical outcome. AB - PURPOSE: Fibroblast growth factor receptor 3 (FGFR3) mutations were recently found at a high frequency in well-differentiated urothelial cell carcinoma (UCC). We investigated the relationship between FGFR3 status and three molecular markers (MIB-1, P53, and P27kip1) associated with worse prognosis and determined the reproducibility of pathologic grade and molecular variables. PATIENTS AND METHODS: In this multicenter study, we included 286 patients with primary (first diagnosis) UCC. The histologic slides were reviewed. FGFR3 status was examined by polymerase chain reaction-single strand conformation polymorphism and sequencing. Expression levels of MIB-1, P53, and P27kip1 were determined by immunohistochemistry. Mean follow-up was 5.5 years (range, 0.4 to 18.4 years). RESULTS: FGFR3 mutations were detected in 172 (60%) of 286 UCCs. Grade 1 tumors had an FGFR3 mutation in 88% of patient samples and grade 3 tumors in 16% of patient samples. Conversely, aberrant expression patterns of MIB-1, P53, and P27kip1 were seen in 5%, 2%, and 3% of grade 1 tumors and in 85%, 60%, and 56% of grade 3 tumors, respectively. In multivariate analysis with recurrence rate, progression, and disease-specific survival as end points, the combination of FGFR3 and MIB-1 proved independently significant in all three cases. By using these two molecular markers, three molecular grades (mGs) could be identified: mG1 (mutation; normal expression), favorable prognosis; mG2 (two remaining combinations), intermediate prognosis; and mG3 (no mutation; high expression), poor prognosis. The molecular variables were more reproducible than pathologic grade (85% to 100% v 47% to 61%). CONCLUSION: The FGFR3 mutation represents the favorable molecular pathway of UCC. Molecular grading provides a new, simple, and highly reproducible tool for clinical decision making in UCC patients. PMID- 12743145 TI - Quality-of-care indicators for early-stage prostate cancer. AB - PURPOSE: Decisions regarding treatment for early-stage prostate cancer are frustrated not only by inadequate evidence favoring one treatment modality but also by the absence of data comparing quality among providers. In fact, the choice of provider may be as important as the choice of treatment. We undertook this study to develop an infrastructure to evaluate variations in quality of care for men with early-stage prostate cancer. METHODS: We enlisted several sources to develop a list of proposed quality-of-care indicators and covariates. After an extensive structured literature review and a series of focus groups with patients and their spouses, we conducted structured interviews with national academic leaders in prostate cancer treatment. We then convened an expert panel using the RAND consensus method to discuss and rate the validity and feasibility of the proposed quality indicators and covariates. RESULTS: The panel endorsed 49 quality-of-care indicators and 14 covariates, which make up our final list of candidate measures. Several domains of quality are represented in the selected indicators, including patient volume, pretreatment referrals, preoperative testing, interpretation of pathology specimens, and 10-year disease-free survival. Covariates include measures of case-mix, such as patient age and comorbidity. CONCLUSION: This study establishes a foundation on which to build quality-of-care assessment tools to evaluate the treatment of early-stage prostate cancer. The next step is to field-test the indicators for feasibility, reliability, validity, and clinical utility in a population-based sample. This work will begin to inform medical decision-making for patients and their physicians. PMID- 12743146 TI - Fluoxetine versus placebo in advanced cancer outpatients: a double-blinded trial of the Hoosier Oncology Group. AB - PURPOSE: To determine whether fluoxetine improves overall quality of life (QOL) in advanced cancer patients with symptoms of depression revealed by a simple survey. PATIENTS AND METHODS: One hundred sixty-three patients with an advanced solid tumor and expected survival between 3 and 24 months were randomly assigned in a double-blinded fashion to receive either fluoxetine (20 mg daily) or placebo for 12 weeks. Patients were screened for at least minimal depressive symptoms and assessed every 3 to 6 weeks for QOL and depression. Patients with recent exposure to antidepressants were excluded. RESULTS: The groups were comparable at baseline in terms of age, sex, disease distribution, performance status, and level of depressive symptoms. One hundred twenty-nine patients (79%) completed at least one follow-up assessment. Analysis using generalized estimating equation modeling revealed that patients treated with fluoxetine exhibited a significant improvement in QOL as shown by the Functional Assessment of Cancer Therapy General, compared with patients given placebo (P =.01). Specifically, the level of depressive symptoms expressed was lower in patients treated with fluoxetine (P =.0005), and the subgroup of patients showing higher levels of depressive symptoms on the two-question screening survey were the most likely to benefit from treatment. CONCLUSION: In this mix of patients with advanced cancer who had symptoms of depression as determined by a two-question bedside survey, use of fluoxetine was well tolerated, overall QOL was improved, and depressive symptoms were reduced. PMID- 12743147 TI - Quality of life in a randomized trial of group psychosocial support in metastatic breast cancer: overall effects of the intervention and an exploration of missing data. AB - PURPOSE: To evaluate the effect of a standardized group psychosocial intervention on health-related quality of life (HrQOL) in women with metastatic breast cancer and to explore the effect of missing data in HrQOL analyses. PATIENTS AND METHODS: Between 1993 and 1998, seven Canadian centers randomly assigned 235 eligible women to participate in a weekly, 90-minute, therapist-led support group that adhered to principles of supportive-expressive (SE) therapy or to a control arm (no SE). All women received educational material and any type of medical or psychosocial care deemed necessary. HrQOL data were prospectively collected using the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire C30 (EORTC QLQ-C30) at baseline, 4, 8, and 12 months. The primary HrQOL analyses compared scores in the two study arms. Analyses were limited to women with appropriate baseline HrQOL information (n = 215). RESULTS: Baseline EORTC QLQ-C30 scores were not different between the two study arms (all P >.05). Primary analysis of all subscales failed to show a significant influence of the intervention on HrQOL (all P >.05). There was a significant deterioration over time in several functional scales of the EORTC QLQ-C30: global (P =.03), physical (P =.0002), role (P =.01), and cognitive functioning (P =.04); and in symptom scales: dyspnea (P =.007), appetite loss (P =.04), and fatigue (P =.003); these changes were independent of randomization allocation. Results were similar in additional analyses of overall HrQOL using a variety of approaches to handling missing data. CONCLUSION: Supportive-expressive group therapy in patients with metastatic breast cancer does not appear to influence HrQOL, as measured by the EORTC QLQ-C30. PMID- 12743148 TI - C-kit receptor expression in Ewing's sarcoma: lack of prognostic value but therapeutic targeting opportunities in appropriate conditions. AB - PURPOSE: Autocrine/paracrine stimulation of c-kit has been recently observed in Ewing's sarcoma (ES) cell lines. In this study, we tested the prognostic and therapeutic role of the receptor in this tumor. METHODS: One hundred one ES tumor biopsies were evaluated for the expression of c-kit by the avidin-biotin peroxidase procedure. Effectiveness of STI-571 (Gleevec; Novartis, Basel, Switzerland), a selective inhibitor of specific tyrosine kinases, was analyzed with respect to in vitro growth and migration inhibition, as single agent or in combination with doxorubicin. RESULTS: Approximately 30% of patients expressed c kit in their primary tumors. No significant association between the expression of the receptor and the clinical outcome was observed. In vitro growth of ES cell lines showing high levels of c-kit demonstrated limited inhibition by exposure to STI-571 (10 micromol/L is required to obtain 40% to 50% of growth inhibition). A decrease of stem-cell factor-mediated ES cell migration was also found. The drug acted additively with doxorubicin in inhibiting ES cell growth. CONCLUSION: The negative prognostic findings and the limited in vitro therapeutic activity of STI 571 indicate that the putative aberrant signaling provided by c-kit overexpression may be dispensable for ES development and unlikely to constitute a critical therapeutic target. Accordingly, the dose of STI-571 required to give a significant ES growth inhibition is much higher than for those tumors in which mutations of c-kit constitute a relevant pathogenetic event. Nevertheless, in the subset of ES patients showing a high level of c-kit expression, the activity of the drug may be exploited in combination with standard therapy. PMID- 12743149 TI - Adiposity and sex hormones in postmenopausal breast cancer survivors. AB - PURPOSE: Overweight and obese women with breast cancer have poorer survival compared with thinner women. One possible reason is that breast cancer survivors with higher degrees of adiposity have higher concentrations of tumor-promoting hormones. This study examined the association between adiposity and concentrations of estrogens, androgens, and sex hormone-binding globulin (SHBG) in a population-based sample of postmenopausal women with breast cancer. METHODS: We studied the associations between body mass index (BMI), body fat mass, and percent body fat, measured by dual-energy x-ray absorptiometry scan, waist circumference, and waist-to-hip circumference ratio, with concentrations of estrone, estradiol, testosterone, SHBG, dehydroepiandrosterone sulfate, free estradiol, and free testosterone in 505 postmenopausal women in western Washington and New Mexico with incident stage 0 to IIIA breast cancer. Blood and adiposity measurements were performed between 4 and 12 months after diagnosis. RESULTS: Obese women (BMI > or = 30) had 35% higher concentrations of estrone and 130% higher concentrations of estradiol compared with lighter-weight women (BMI < 22.0; P =.005 and.002, respectively). Similar associations were observed for body fat mass, percent body fat, and waist circumference. Testosterone concentrations also increased with increasing levels of adiposity (P =.0001). Concentrations of free estradiol and free testosterone were two to three times greater in overweight and obese women compared with lighter-weight women (P =.0001). CONCLUSION: These data provide information about potential hormonal explanations for the association between adiposity and breast cancer prognosis. These sex hormones may be useful biomarkers for weight loss intervention studies in women with breast cancer. PMID- 12743150 TI - Serum HER-2/neu and response to the aromatase inhibitor letrozole versus tamoxifen. AB - PURPOSE: To determine the effect of elevated serum HER-2/neu on the response of metastatic breast cancer patients to an aromatase inhibitor versus an antiestrogen. PATIENTS AND METHODS: Five hundred sixty-two estrogen receptor positive metastatic breast cancer patients were randomized to first-line hormone therapy with either letrozole or tamoxifen. An automated enzyme-linked immunosorbent assay was used to detect serum HER-2/neu. RESULTS: For patients with normal serum HER-2/neu (70.5%), objective response rate (ORR; 39% in letrozole-treated patients v 26% in tamoxifen-treated patients; P =.008), clinical benefit (CB; 57% v 45%; P =.016), time to progression (TTP; median, 12.2 v 8.5 months; P =.0019), and time to treatment failure (TTF; median, 11.6 v 6.2 months; P =.0066) were significantly better in patients treated with letrozole. In the elevated HER-2/neu group (29.5%), there was no significant difference in ORR (17% in letrozole-treated patients v 13% in tamoxifen-treated patients; P =.45) or CB (33% v 26%; P =.31), but there was a strong trend in favor of a longer TTP with letrozole (median, 6.1 v 3.3 months; P =.0596) and a significantly longer TTF with letrozole (median, 6.0 v 3.2 months; P =.0418). Multivariate analysis revealed that elevated serum HER-2/neu was a negative predictor for ORR and TTP. CONCLUSION: Patients with normal serum HER-2/neu receiving letrozole demonstrated a significantly greater ORR and CB and longer TTP and TTF than patients receiving tamoxifen. Although in patients with elevated serum HER-2/neu there was no significant difference between letrozole and tamoxifen in ORR or CB, there was a strong trend favoring longer TTP and significantly longer TTF with letrozole. PMID- 12743151 TI - Progesterone receptor status significantly improves outcome prediction over estrogen receptor status alone for adjuvant endocrine therapy in two large breast cancer databases. AB - PURPOSE: To determine whether progesterone receptor (PgR) status provides additional value to estrogen receptor (ER) status and improves prediction of benefit from endocrine treatment among patients with primary breast cancer. PATIENTS AND METHODS: Clinical outcomes of patients in two large databases were analyzed as a function of steroid receptor status. The first database (PP), contained 3,739 patients who did not receive any systemic adjuvant therapy and 1,688 patients who received adjuvant endocrine therapy but no chemotherapy. The second database (SPORE), contained 10,444 patients who received adjuvant endocrine therapy but no chemotherapy. Biochemical ER and PgR assays were identically performed in two different central laboratories. RESULTS: In univariate and multivariate analyses, the prognostic significance of PgR status among systemically untreated patients is modest. Among endocrine-treated patients, however, multivariate analyses, including lymph-node involvement, tumor size, and age, demonstrate that PgR status is independently associated with disease-free and overall survival. For recurrence, the reduction in relative risk (RR) was 25% for ER-positive/PgR-negative patients and 53% for ER-positive/PgR positive patients, compared with ER-negative/PgR-negative patients (P <.0001, PP patients). Patients with ER-positive/PgR-negative tumors have a reduction in RR of death of 30% (SPORE patients) and 38% (PP patients), compared with patients with ER-negative/PgR-negative tumors (P <.0001). For ER-positive/PgR-positive tumors, the reduction of the risk of death was greater than 46% in SPORE patients and 58% in PP patients, indicating that ER-positive/PgR-positive patients derive more benefit from endocrine therapy (P <.0001). CONCLUSION: When accurately measured, PgR status is an independent predictive factor for benefit from adjuvant endocrine therapy. Therefore, PgR status should be taken into account when discussing RR reductions expected from endocrine treatment with individual patients. PMID- 12743152 TI - Phase II trial of ZD1839 in recurrent or metastatic squamous cell carcinoma of the head and neck. AB - PURPOSE: The epidermal growth factor receptor (EGFR) is a mediator of squamous cell carcinoma of the head and neck (SCCHN) development. ZD1839 is an orally active, selective EGFR tyrosine kinase inhibitor. This phase II study sought to explore the activity, toxicity, and pharmacodynamics of ZD1839 in SCCHN. PATIENTS AND METHODS: Patients with recurrent or metastatic SCCHN were enrolled through the University of Chicago Phase II Consortium. Patients were allowed no more than one prior therapy for recurrent or metastatic disease and were treated with single-agent ZD1839 500 mg/d. Patient tumor biopsies were obtained and stained immunohistochemically for EGFR, extracellular signal-regulated kinase 1 (ERK1), and phosphorylated ERK1 (p-ERK). Study end points included response rate, time to progression, median survival, and inhibition of p-ERK. RESULTS: Fifty-two patients were enrolled (40 male and 12 female) with a median age of 59 years (range, 34 to 84 years). Fourteen patients received ZD1839 through a feeding tube. Half the cohort received ZD1839 as second-line therapy. Forty-seven patients were assessable for response, with an observed response rate of 10.6% and a disease control rate of 53%. Median time to progression and survival were 3.4 and 8.1 months, respectively. The only grade 3 toxicity encountered was diarrhea in three patients. Performance status and development of skin toxicity were found to be strong predictors of response, progression, and survival. Ten biopsy samples were assessable and revealed no significant change in EGFR or p ERK expression with ZD1839 therapy. CONCLUSION: ZD1839 has single-agent activity and is well tolerated in refractory SCCHN. In contrast to other reports, development of skin toxicity was a statistically significant predictor of response and improved outcome. PMID- 12743153 TI - Significant correlation between the degree of WT1 expression and the International Prognostic Scoring System Score in patients with myelodysplastic syndromes. AB - PURPOSE: To determine whether pattern of WT1 gene expression is a useful marker for establishing prognosis and tracking disease progression in patients with myelodysplastic syndromes (MDS). PATIENTS AND METHODS: We performed a quantitative assessment of the WT1 transcript amount by real-time quantitative polymerase chain reaction (RQ-PCR) in 173 samples (131 bone marrow samples and 42 peripheral-blood samples) from 131 patients with MDS (79 patients with refractory anemia [RA], 31 with RA with excess blasts [RAEB], 18 with secondary acute myeloid leukemia [s-AML] evolved from MDS, and three with deletion of 5q as the sole cytogenetic abnormality). Values obtained were correlated with the blast percentage and International Prognostic Scoring System (IPSS) score. RESULTS: Sixty-five percent of BM and 78% of PB samples for RA and 100% of BM and PB samples of RAEB and s-AML expressed WT1 transcript amounts greater than the level observed in healthy volunteers. The degree of WT1 expression was highly correlated with the type of MDS, was much higher in RAEB and s-AML compared with RA, and increased during disease progression. Moreover, a significant correlation was found between WT1 expression levels, blast cell percentage, and the presence of cytogenetic abnormalities. Therefore, we found a significant correlation between the amount of WT1 transcripts and the IPSS score, which currently represents the most reliable risk index of disease progression available for MDS patients. CONCLUSION: WT1 is a useful molecular marker for risk assessment in MDS patients. PMID- 12743154 TI - Combination of fludarabine and mitoxantrone in untreated stages III and IV low grade lymphoma: S9501. AB - PURPOSE: To determine the efficacy of combination fludarabine and mitoxantrone (FN) in untreated stages III and IV low-grade lymphoma. The major end point was to estimate progression-free survival (PFS) in all eligible patients. PATIENTS AND METHODS: Seventy-eight eligible patients were registered. Chemotherapy courses were administered every 4 weeks with mitoxantrone 10 mg/m2 on day 1 and fludarabine 25 mg/m2 on days 1, 2, and 3 for a total of six to eight cycles. Pneumocystis carinii prophylaxis was required. RESULTS: Seventy-three patients (94%) attained an objective response. Complete remission was demonstrated in 34 patients (44%) and partial remission was demonstrated in 39 patients (50%). With a median follow-up time of 5.5 years, the median PFS was 32 months, with a 4-year PFS rate of 38%. Median survival has not been reached and 88% of all patients are alive at 4 years. The application of the International Prognostic Index and serologic staging showed significant differences in PFS in all risk groups, whereas overall survival was markedly worse for the highest-risk group in either prognostic model. Three prior Southwest Oncology Group trials using a regimen of cyclophosphamide, doxorubicin, vincristine, and prednisone or a combination of prednisone, vincristine, methotrexate, cytarabine, cyclophosphamide, etoposide, nitrogen mustard, vincristine, procarbazine, and prednisone in similar patient populations demonstrated comparable clinical outcome, although the 4-year survival for FN was better. FN was well tolerated, but mild to severe reversible myelosuppression was noted. Other complications were rare. CONCLUSION: FN is an effective, safe chemotherapy combination for patients with advanced-stage, low grade lymphoma. Clinical outcomes were comparable to prior published data using anthracycline-based regimens. PMID- 12743156 TI - Primary metastatic osteosarcoma: presentation and outcome of patients treated on neoadjuvant Cooperative Osteosarcoma Study Group protocols. AB - PURPOSE: To determine demographic data and define prognostic factors for long term outcome in patients presenting with high-grade osteosarcoma of bone with clinically detectable metastases at initial presentation. PATIENTS AND METHODS: Of 1,765 patients with newly diagnosed, previously untreated high-grade osteosarcomas of bone registered in the neoadjuvant Cooperative Osteosarcoma Study Group studies before 1999, 202 patients (11.4%) had proven metastases at diagnosis and therefore were enrolled onto an analysis of demographic-, tumor-, and treatment-related variables, response, and survival. The intended therapeutic strategy included pre- and postoperative multiagent chemotherapy as well as aggressive surgery of all resectable lesions. RESULTS: With a median follow-up of 1.9 years (5.5 years for survivors), 60 patients were alive, 37 of whom were in continuously complete surgical remission. Actuarial overall survival rates at 5 and 10 (same value for 15) years were 29% (SE = 3%) and 24% (SE = 4%), respectively. In univariate analysis, survival was significantly correlated with patient age, site of the primary tumor, number and location of metastases, number of involved organ systems, histologic response of the primary tumor to preoperative chemotherapy, and completeness and time point of surgical resection of all tumor sites. However, after multivariate Cox regression analysis, only multiple metastases at diagnosis (relative hazard rate [RHR] = 2.3) and macroscopically incomplete surgical resection (RHR = 2.4) remained significantly associated with inferior outcomes. CONCLUSION: The number of metastases at diagnosis and the completeness of surgical resection of all clinically detected tumor sites are of independent prognostic value in patients with proven primary metastatic osteosarcoma. PMID- 12743155 TI - Consolidation docetaxel after concurrent chemoradiotherapy in stage IIIB non small-cell lung cancer: phase II Southwest Oncology Group Study S9504. AB - PURPOSE: To test the concept of taxane sequencing in combined-modality therapy, this phase II trial (S9504) evaluated consolidation docetaxel after concurrent chemoradiotherapy in patients with pathologically documented stage IIIB non-small cell lung cancer (NSCLC). Results were compared with those of the predecessor study (S9019) with identical eligibility, staging criteria, and treatment, excepting docetaxel consolidation. PATIENTS AND METHODS: Treatment consisted of cisplatin 50 mg/m2 on days 1, 8, 29, and 36, etoposide 50 mg/m2 on days 1 through 5 and 29 through 33, and concurrent thoracic radiotherapy (total dose of 61 Gy). Consolidation docetaxel started 4 to 6 weeks after chemoradiotherapy at an initial dose of 75 mg/m2. RESULTS: Stage subsets (tumor-node-metastasis system) in 83 eligible patients were as follows: T4N0/1, 31 patients (37%); T4N2, 22 patients (27%), and T1-3N3, 30 patients (36%). Concurrent chemoradiotherapy was generally well tolerated, but two patients died from probable radiation associated pneumonitis. Neutropenia during consolidation docetaxel was common (57% with grade 4) and most frequent during escalation to 100 mg/m2. Median progression-free survival was 16 months, median survival was 26 months, and 1-, 2 , and 3-year survival rates were 76%, 54%, and 37%, respectively. Brain metastasis was the most common site of failure. In S9019, median survival was 15 months and 1-, 2-, and 3-year survival rates were 58%, 34%, and 17%, respectively. CONCLUSION: Consolidation docetaxel after concurrent chemoradiotherapy in stage IIIB NSCLC is feasible and generally tolerable, and results compare favorably with the predecessor trial S9019. Nevertheless, this study remains hypothesis-generating and does not provide definitive evidence of the benefit of this approach. Phase III trials evaluating the S9504 regimen have been initiated to validate these results. PMID- 12743157 TI - Treatment of intraocular retinoblastoma with vincristine and carboplatin. AB - PURPOSE: To evaluate the efficacy of chemoreduction using vincristine and carboplatin in preventing or delaying external-beam radiotherapy (EBRT) or enucleation in patients with intraocular retinoblastoma. PATIENTS AND METHODS: Twenty-five patients (43 eyes) with newly diagnosed intraocular retinoblastoma received primary treatment with eight courses of vincristine and carboplatin. Focal treatments were delayed until documentation of disease progression. Outcome measures for each eye were length of time to disease progression, avoidance or delay of EBRT, and globe survival. Event-free survival was defined as the length of time to EBRT or enucleation. RESULTS: Disease in all eyes responded to chemotherapy and progressed in only two patients before completion of the eight courses of therapy. Disease in all but four eyes progressed and required focal treatments. Event-free survival estimates at 2 years were 59.2% +/- 12.0% for Reese-Ellsworth group I, II, and III eyes and 26.3% +/- 9.2% for group IV and V eyes. Nineteen eyes (44.2%) required EBRT and 13 eyes (30.2%) were enucleated. The ocular salvage rate was 83.3% for Reese-Ellsworth group I to III eyes and 52.6% for group IV and V eyes. For those patients receiving EBRT, the median time from enrollment to EBRT was 9.5 months (median age at EBRT, 21 months). CONCLUSION: In combination with appropriate early intensive focal treatments, chemoreduction with vincristine and carboplatin, without etoposide, may be an alternative treatment for patients with early-stage intraocular retinoblastoma, although additional studies are needed. Patients with advanced intraocular disease require more aggressive treatments. PMID- 12743158 TI - Prognostic factors for children with Hodgkin's disease treated with combined modality therapy. AB - PURPOSE: Evaluation of pretreatment factors to identify children at high risk for relapse after combined-modality therapy for Hodgkin's disease. PATIENTS AND METHODS: From 1990 to 2000, 328 pediatric patients with clinical stage I to IV Hodgkin's disease were treated with chemotherapy and low-dose involved-field radiotherapy on prospective, collaborative, risk-adapted protocols at three institutions. Pretreatment factors were analyzed by univariate and multivariate analysis for prognostic significance for 5-year disease-free survival (DFS) and overall survival (OS). RESULTS: With a median follow-up of 59 months (range, 8 to 125 months), the 5-year DFS and OS for all patients were 83% and 93%, respectively. Several factors were associated with inferior DFS and OS by univariate analysis. By multivariate analysis, male sex; stage IIB, IIIB, or IV disease; bulky mediastinal disease; WBC more than 13.5 x 10(3)/mm3; and hemoglobin less than 11.0 g/dL were significant for inferior DFS. A prognostic index was developed incorporating the five significant factors from the multivariate analysis, assigning each a score of 1. The 5-year DFS and OS for children with a prognostic score of 0 to 1 were 94% and 99%; score 2, 85% and 96%; score 3, 71% and 92%; and score 4 or 5, 49% and 72%, respectively. There was a significant difference in DFS among each of these groups, with significantly worse OS in those with a score of 4 to 5. CONCLUSION: A prognostic index that was based on five pretreatment factors correlated with inferior DFS by multivariate analysis stratified patients by outcome; this may be useful in assigning children with Hodgkin's disease to risk-adapted therapy. PMID- 12743159 TI - Role of telomeres and telomerase in the pathogenesis of human cancer. AB - Specialized nucleoprotein structures, termed telomeres, cap the ends of human chromosomes. These terminal structures, composed of repetitive arrays of guanine rich hexameric DNA together with specific telomere-binding proteins, play essential roles in protecting the chromosome from damage and degradation. In addition, several lines of evidence implicate telomere maintenance as an important regulator of cell life span. Activation of telomerase, a dedicated reverse transcriptase that synthesizes telomeric sequences, is strongly associated with cancer, and recent observations confirm that telomeres and telomerase perform important roles in both suppressing and facilitating malignant transformation. These dual functions of telomere biology are evident in the clinical manifestations of the multisystem syndrome, dyskeratosis congenita, forms of which display defects in telomerase function. Recent advances in our understanding of telomere biology indicate that the manipulation of telomeres and telomerase will lead to clinically significant applications in the diagnosis, prevention, and treatment of neoplastic disease. PMID- 12743160 TI - Simple method to eliminate the risk of inadvertent intrathecal vincristine administration. PMID- 12743164 TI - Overall survival is not a realistic end point for clinical trials of new drugs in advanced solid tumors: a critical assessment based on recently reported phase III trials in colorectal and breast cancer. PMID- 12743162 TI - Calcitriol and alopecia--is it the hair apparent? PMID- 12743165 TI - Potentiated L-type Ca2+ channels rectify. AB - Strong depolarization and dihydropyridine agonists potentiate inward currents through native L-type Ca2+ channels, but the effect on outward currents is less clear due to the small size of these currents. Here, we examined potentiation of wild-type alpha1C and two constructs bearing mutations in conserved glutamates in the pore regions of repeats II and IV (E2A/E4A-alpha1C) or repeat III (E3K alpha1C). With 10 mM Ca2+ in the bath and 110 mM Cs+ in the pipette, these mutated channels, expressed in dysgenic myotubes, produced both inward and outward currents of substantial amplitude. For both the wild-type and mutated channels, we observed strong inward rectification of potentiation: strong depolarization had little effect on outward tail currents but caused the inward tail currents to be larger and to decay more slowly. Similarly, exposure to DHP agonist increased the amplitude of inward currents and decreased the amplitude of outward currents through both E2A/E4A-alpha1C and E3K-alpha1C. As in the absence of drug, strong depolarization in the presence of dihydropyridine agonist had little effect on outward tail currents but increased the amplitude and slowed the decay of inward tail currents. We tested whether cytoplasmic Mg2+ functions as the blocking particle responsible for the rectification of potentiated L-type Ca2+ channels. However, even after complete removal of cytoplasmic Mg2+, (-)BayK 8644 still potentiated inward current and partially blocked outward current via E2A/E4A-alpha1C. Although zero Mg2+ did not reveal potentiation of outward current by DHP agonist, it did have two striking effects, (a) a strong suppression of decay of both inward and outward currents via E2A/E4A-alpha1C and (b) a nearly complete elimination of depolarization-induced potentiation of inward tail currents. These results can be explained by postulating that potentiation exposes a binding site in the pore to which an intracellular blocking particle can bind and produce inward rectification of the potentiated channels. PMID- 12743166 TI - Pore structure influences gating properties of the T-type Ca2+ channel alpha1G. AB - The selectivity filter of all known T-type Ca2+ channels is built by an arrangement of two glutamate and two aspartate residues, each one located in the P-loops of domains I-IV of the alpha1 subunit (EEDD locus). The mutations of the aspartate residues to glutamate induce changes in the conduction properties, enhance Cd2+ and proton affinities, and modify the activation curve of the channel. Here we further analyze the role of the selectivity filter in the gating mechanisms of T-type channels by comparing the kinetic properties of the alpha1G subunit (CaV3.1) to those of pore mutants containing aspartate-to-glutamate substitution in domains III (EEED) or IV (EEDE). The change of the extracellular pH induced similar effects on the activation properties of alpha1G and both pore mutants, indicating that the larger affinity of the mutant channels for protons is not the cause of the gating modifications. Both mutants showed alterations in several gating properties with respect to alpha1G, i.e., faster macroscopic inactivation in the voltage range from -10 to 50 mV, positive voltage shift and decrease in the voltage sensitivity of the time constants of activation and deactivation, decrease of the voltage sensitivity of the steady-state inactivation, and faster recovery from inactivation for long repolarization periods. Kinetic modeling suggests that aspartate-to-glutamate mutations in the EEDD locus of alpha1G modify the movement of the gating charges and alter the rate of several gating transitions. These changes are independent of the alterations of the selectivity properties and channel protonation. PMID- 12743168 TI - An anionic ryanoid, 10-O-succinoylryanodol, provides insights into the mechanisms governing the interaction of ryanoids and the subsequent altered function of ryanodine-receptor channels. AB - We have investigated the interactions of a novel anionic ryanoid, 10-O succinoylryanodol, with individual mammalian cardiac muscle ryanodine receptor channels under voltage clamp conditions. As is the case for all ryanoids so far examined, the interaction of 10-O-succinoylryanodol with an individual RyR channel produces profound alterations in both channel gating and rates of ion translocation. In the continued presence of the ryanoid the channel fluctuates between periods of normal and modified gating, indicating a reversible interaction of the ligand with its receptor. Unlike the majority of ryanoids, we observe a range of different fractional conductance states of RyR in the presence of 10-O-succinoylryanodol. We demonstrate that 10-O-succinoylryanodol is a very flexible molecule and propose that each fractional conductance state arises from the interaction of a different conformer of the ryanoid molecule with the RyR channel. The probability of channel modification by 10-O-succinoylryanodol is dependent on the transmembrane holding potential. Comparison of the voltage dependence of channel modification by this novel anionic ryanoid with previous data obtained with cationic and neutral ryanoids reveals that the major influence of transmembrane potential on the probability of RyR channel modification by ryanoids results from an alteration in receptor affinity. These investigations also demonstrate that the charge of the ryanoid has a major influence on the rate of association of the ligand with its receptor indicating that ionic interactions are likely to be involved in this reaction. PMID- 12743167 TI - Extracellular Ca2+ modulates the effects of protons on gating and conduction properties of the T-type Ca2+ channel alpha1G (CaV3.1). AB - Since Ca2+ is a major competitor of protons for the modulation of high voltage activated Ca2+ channels, we have studied the modulation by extracellular Ca2+ of the effects of proton on the T-type Ca2+ channel alpha1G (CaV3.1) expressed in HEK293 cells. At 2 mM extracellular Ca2+ concentration, extracellular acidification in the pH range from 9.1 to 6.2 induced a positive shift of the activation curve and increased its slope factor. Both effects were significantly reduced if the concentration was increased to 20 mM or enhanced in the absence of Ca2+. Extracellular protons shifted the voltage dependence of the time constant of activation and decreased its voltage sensitivity, which excludes a voltage dependent open pore block by protons as the mechanism modifying the activation curve. Changes in the extracellular pH altered the voltage dependence of steady state inactivation and deactivation kinetics in a Ca2+-dependent manner, but these effects were not strictly correlated with those on activation. Model simulations suggest that protons interact with intermediate closed states in the activation pathway, decreasing the gating charge and shifting the equilibrium between these states to less negative potentials, with these effects being inhibited by extracellular Ca2+. Extracellular acidification also induced an open pore block and a shift in selectivity toward monovalent cations, which were both modulated by extracellular Ca2+ and Na+. Mutation of the EEDD pore locus altered the Ca2+-dependent proton effects on channel selectivity and permeation. We conclude that Ca2+ modulates T-type channel function by competing with protons for binding to surface charges, by counteracting a proton-induced modification of channel activation and by competing with protons for binding to the selectivity filter of the channel. PMID- 12743169 TI - Determinant of HIV-1 mutational escape from cytotoxic T lymphocytes. AB - CD8+ class I-restricted cytotoxic T lymphocytes (CTLs) usually incompletely suppress HIV-1 in vivo, and while analogous partial suppression induces antiretroviral drug-resistance mutations, epitope escape mutations are inconsistently observed. However, escape mutation depends on the net balance of selective pressure and mutational fitness costs, which are poorly understood and difficult to study in vivo. Here we used a controlled in vitro system to evaluate the ability of HIV-1 to escape from CTL clones, finding that virus replicating under selective pressure rapidly can develop phenotypic resistance associated with genotypic changes. Escape varied between clones recognizing the same Gag epitope or different Gag and RT epitopes, indicating the influence of the T cell receptor on pressure and fitness costs. Gag and RT escape mutations were monoclonal intra-epitope substitutions, indicating limitation by fitness constraints in structural proteins. In contrast, escape from Nef-specific CTL was more rapid and consistent, marked by a polyclonal mixture of epitope point mutations and upstream frameshifts. We conclude that incomplete viral suppression by CTL can result in rapid emergence of immune escape, but the likelihood is strongly determined by factors influencing the fitness costs of the particular epitope targeted and the ability of responding CTL to recognize specific epitope variants. PMID- 12743170 TI - Aberrant chemokine receptor expression and chemokine production by Langerhans cells underlies the pathogenesis of Langerhans cell histiocytosis. AB - Langerhans cell histiocytosis (LCH) is characterized by a clonal proliferation and retention of cells with a Langerhans cell (LC)-like phenotype at various sites within the body. The present study set out to elucidate whether aberrant expression of chemokine receptors or dysregulation of chemokine production in LCH lesions could explain abnormal retention of these cells. Immunohistochemical analysis on 13 LCH biopsies of bone, skin, and lymph node all expressed the immature dendritic cell (DC) marker CCR6 on the lesional LCs and absence of the mature DC marker CCR7. Furthermore, regardless of the tissue site, LCH lesions markedly overexpressed CCL20/MIP-3alpha, the ligand for CCR6. The lesional LCs appeared to be the source of this CCL20/MIP-3alpha production as well as other inflammatory chemokines such as CCL5/RANTES and CXCL11/I-TAC. These may explain the recruitment of eosinophils and CD4+CD45RO+ T cells commonly found in LCH lesions. The findings of this study emphasize that, despite abundant TNF-alpha, lesional LCs remain in an immature state and are induced to produce chemokines, which via autocrine and paracrine mechanisms cause not only the retention of the lesional LCs but also the recruitment and retention of other lesional cells. We postulate that the lesional LCs themselves control the persistence and progression of LCH. PMID- 12743171 TI - A human minor histocompatibility antigen resulting from differential expression due to a gene deletion. AB - Minor histocompatibility antigens (minor H antigens) are targets of graft-versus host disease and graft-versus-leukemia responses after allogeneic human leukocyte antigen identical hematopoietic stem cell transplantation. Only a few human minor H antigens have been molecularly characterized and in all cases, amino acid differences between homologous donor and recipient proteins due to nucleotide polymorphisms in the respective genes were responsible for immunogenicity. Here, we have used cDNA expression cloning to identify a novel human minor H antigen encoded by UGT2B17, an autosomal gene in the multigene UDP-glycosyltransferase 2 family that is selectively expressed in liver, intestine, and antigen-presenting cells. In contrast to previously defined human minor H antigens, UGT2B17 is immunogenic because of differential expression of the protein in donor and recipient cells as a consequence of a homozygous gene deletion in the donor. Deletion of individual members of large gene families is a common form of genetic variation in the population and our results provide the first evidence that differential protein expression as a consequence of gene deletion is a mechanism for generating minor H antigens in humans. PMID- 12743173 TI - An essential role of cytosolic phospholipase A2alpha in prostaglandin E2-mediated bone resorption associated with inflammation. AB - Prostaglandin E (PGE)2 produced by osteoblasts acts as a potent stimulator of bone resorption. Inflammatory bone loss is accompanied by osteoclast formation induced by bone-resorbing cytokines, but the mechanism of PGE2 production and bone resorption in vivo is not fully understood. Using cytosolic phospholipase A2alpha (cPLA2alpha)-null mice, we examined the role of cPLA2alpha in PGE2 synthesis and bone resorption. In bone marrow cultures, interleukin (IL)-1 markedly stimulated PGE2 production and osteoclast formation in wild-type mice, but not in cPLA2alpha-null mice. Osteoblastic bone marrow stromal cells induced the expression of cyclooxygenase (COX)-2 and membrane-bound PGE2 synthase (mPGES) in response to IL-1 and lipopolysaccharide (LPS) to produce PGE2. Osteoblastic stromal cells collected from cPLA2alpha-null mice also induced the expression of COX-2 and mPGES by IL-1 and LPS, but could not produce PGE2 due to the lack of arachidonic acid release. LPS administration to wild-type mice reduced femoral bone mineral density by increased bone resorption. In cPLA2alpha-null mice, however, LPS-induced bone loss could not be observed at all. Here, we show that cPLA2alpha plays a key role in PGE production by osteoblasts and in osteoclastic bone resorption, and suggest a new approach to inflammatory bone disease by inhibiting cPLA2alpha. PMID- 12743172 TI - Cytosolic phospholipase A2alpha-deficient mice are resistant to collagen-induced arthritis. AB - Pathogenic mechanisms relevant to rheumatoid arthritis occur in the mouse model of collagen-induced arthritis (CIA). Cytosolic phospholipase A2alpha (cPLA2alpha) releases arachidonic acid from cell membranes to initiate the production of prostaglandins and leukotrienes. These inflammatory mediators have been implicated in the development of CIA. To test the hypothesis that cPLA2alpha plays a key role in the development of CIA, we backcrossed cPLA2alpha-deficient mice on the DBA/1LacJ background that is susceptible to CIA. The disease severity scores and the incidence of disease were markedly reduced in cPLA2alpha-deficient mice compared with wild-type littermates. At completion of the study, >90% of the wild-type mice had developed disease whereas none of the cPLA2alpha-deficient mice had more than one digit inflamed. Furthermore, visual disease scores correlated with severity of disease determined histologically. Pannus formation, articular fibrillation, and ankylosis were all dramatically reduced in the cPLA2alpha-deficient mice. Although the disease scores differed significantly between cPLA2alpha mutant and wild-type mice, anti-collagen antibody levels were similar in the wild-type mice and mutant littermates. These data demonstrate the critical role of cPLA2alpha in the pathogenesis of CIA. PMID- 12743177 TI - Caring for dying patients: what is right? PMID- 12743174 TI - Mlh1 can function in antibody class switch recombination independently of Msh2. AB - Mismatch repair proteins participate in antibody class switch recombination, although their roles are unknown. Previous nucleotide sequence analyses of switch recombination junctions indicated that the roles of Msh2 and the MutL homologues, Mlh1 and Pms2, differ. We now asked if Msh2 and Mlh1 function in the same pathway during switch recombination. Splenic B cells from mice deficient in both these proteins were induced to undergo switching in culture. The frequency of switching is reduced, similarly to that of B cells singly deficient in Msh2 or Mlh1. However, the nucleotide sequences of the Smu-Sgamma3 junctions resemble junctions from Mlh1- but not from Msh2-deficient cells, suggesting Mlh1 functions either independently of or before Msh2. The substitution mutations within S regions that are known to accompany switch recombination are increased in Msh2- and Mlh1 single-deficient cells and further increased in the double-deficient cells, again suggesting these proteins function independently in class switch recombination. The finding that MMR functions to reduce mutations in switch regions is unexpected since MMR proteins have been shown to contribute to somatic hypermutation of antibody variable region genes. PMID- 12743178 TI - Doc, how much time do I have? PMID- 12743179 TI - A revisitation of "doc, how much time do I have?". PMID- 12743181 TI - Truth or consequences: what to do when the patient doesn't want to know. PMID- 12743182 TI - Discussing do-not-resuscitate status. PMID- 12743180 TI - Tell it like it is. PMID- 12743184 TI - "But doctor, what have I got to lose...?". PMID- 12743183 TI - Discussing do-not-resuscitate status: furthering the discourse. PMID- 12743185 TI - "But doctor, what have I got to lose...?". PMID- 12743186 TI - Discussing hospice care. PMID- 12743187 TI - Tell the children. PMID- 12743188 TI - When the tumor is not the target, tell the children. PMID- 12743189 TI - The power of silence. PMID- 12743190 TI - Overcoming obstacles to hospice care: an ethical examination of inertia and inaction. PMID- 12743191 TI - Sweet time unafflicted. PMID- 12743192 TI - Giving bad news to cancer patients: matching process and content. PMID- 12743193 TI - Death denial. PMID- 12743194 TI - Spirituality and the dying patient. PMID- 12743196 TI - Full-impact medicine. PMID- 12743195 TI - The narrow path. PMID- 12743197 TI - Submitting to autonomy. PMID- 12743199 TI - Just whose autonomy is it? PMID- 12743198 TI - Why me? PMID- 12743200 TI - The right to decide. PMID- 12743201 TI - Looking at the dying patient: the Ferdinand Hodler paintings of Valentine Gode Darel. PMID- 12743202 TI - An existential oncologist. PMID- 12743203 TI - On appetite and its loss. PMID- 12743204 TI - Hazards of quality-of-life data for clinical decision making. PMID- 12743205 TI - To hydrate or not to hydrate: how should it be? PMID- 12743206 TI - Opioid rotation in the management of refractory cancer pain. PMID- 12743207 TI - Interventional pain therapy for intractable abdominal cancer pain. PMID- 12743208 TI - A good death: not just an abstract concept. PMID- 12743209 TI - A "good death" revisited in the context of doctor-patient relationships. PMID- 12743211 TI - House calls by oncologists: a home-visit primer. PMID- 12743210 TI - E-motions: email for written emotional expression. PMID- 12743213 TI - Doc, shouldn't we be getting some tests? PMID- 12743212 TI - Life after breast cancer. PMID- 12743214 TI - Not just tired. PMID- 12743215 TI - Adjuvant chemotherapy for women with young children: our patients as parents. PMID- 12743216 TI - Parents with cancer: who's looking after the children? PMID- 12743217 TI - First, you cry, 25 years later. PMID- 12743218 TI - Caring (really) for patients who use alternative therapies for cancer. PMID- 12743219 TI - The death of idiopathic intracranial hypertension? PMID- 12743220 TI - Vertebrobasilar dissection and cervical spine manipulation A complex pain in the neck. PMID- 12743221 TI - Is seeing believing? Functional imaging of hysterical anesthesia. PMID- 12743222 TI - Bringing epilepsy out of the shadows. PMID- 12743223 TI - Clinical spectrum of succinic semialdehyde dehydrogenase deficiency. AB - Succinic semialdehyde dehydrogenase (SSADH) deficiency is a rare autosomal recessive disorder affecting CNS gamma-aminobutyric acid (GABA) degradation. SSADH, in conjunction with GABA transaminase, converts GABA to succinate. In the absence of SSADH, GABA is converted to 4-OH-butyrate. The presence of 4-OH butyrate, a highly volatile compound, may be undetected on routine organic acid analysis. Urine organic acid testing was modified at the authors' institution in 1999 to screen for the excretion of 4-OH-butyrate by selective ion monitoring gas chromatography-mass spectrometry in addition to total ion chromatography. Since then, five patients with 4-hydroxybutyric aciduria have been identified. The authors add the clinical, neuroimaging, and EEG findings from a new cohort of patients to 51 patients reported in the literature with clinical details. Ages ranged from 1 to 21 years at diagnosis. Clinical findings include mild-moderate mental retardation, disproportionate language dysfunction, hypotonia, hyporeflexia, autistic behaviors, seizures, and hallucinations. Brain MRI performed in five patients at the authors' institution revealed symmetric increased T2 signal in the globus pallidi. SSADH deficiency is an under recognized, potentially manageable neurometabolic disorder. Urine organic acid analysis should include a sensitive method for the detection of 4-hydroxybutyrate and should be obtained from patients with mental retardation or neuropsychiatric disturbance of unknown etiology. PMID- 12743224 TI - Idiopathic intracranial hypertension: the prevalence and morphology of sinovenous stenosis. AB - OBJECTIVE: To determine the prevalence and nature of sinovenous obstruction in idiopathic intracranial hypertension (IIH) using auto-triggered elliptic-centric ordered three-dimensional gadolinium-enhanced MR venography (ATECO MRV). METHODS: In a prospective controlled study, 29 patients with established IIH as well as 59 control patients underwent ATECO MRV. In a randomized blinded fashion, three readers evaluated the images. Using a novel scoring system, each reader graded the degree of stenosis seen in the transverse and sigmoid sinuses of each patient. RESULTS: There was excellent agreement across the three readers for application of the grading system. Substantial bilateral sinovenous stenoses were seen in 27 of 29 patients with IIH and in only 4 of 59 control patients. CONCLUSION: Using ATECO MRV and a novel grading system for quantifying sinovenous stenoses, the authors can identify IIH patients with sensitivity and specificity of 93%. PMID- 12743225 TI - Spinal manipulative therapy is an independent risk factor for vertebral artery dissection. AB - OBJECTIVE: To determine whether spinal manipulative therapy (SMT) is an independent risk factor for cervical artery dissection. METHODS: Using a nested case-control design, the authors reviewed all patients under age 60 with cervical arterial dissection (n = 151) and ischemic stroke or TIA from between 1995 and 2000 at two academic stroke centers. Controls (n = 306) were selected to match cases by sex and within age strata. Cases and controls were solicited by mail, and respondents were interviewed using a structured questionnaire. The medical records of interviewed patients were reviewed by two blinded neurologists to confirm that the patient had stroke or TIA and to determine whether there was evidence of arterial dissection. RESULTS: After interview and blinded chart review, 51 patients with dissection (mean age 41 +/- 10 years; 59% female) and 100 control patients (44 +/- 9 years; 58% female) were studied. In univariate analysis, patients with dissection were more likely to have had SMT within 30 days (14% vs 3%, p = 0.032), to have had neck or head pain preceding stroke or TIA (76% vs 40%, p < 0.001), and to be current consumers of alcohol (76% vs 57%, p = 0.021). In multivariate analysis, vertebral artery dissections were independently associated with SMT within 30 days (OR 6.62, 95% CI 1.4 to 30) and pain before stroke/TIA (OR 3.76, 95% CI 1.3 to 11). CONCLUSIONS: This case controlled study of the influence of SMT and cervical arterial dissection shows that SMT is independently associated with vertebral arterial dissection, even after controlling for neck pain. Patients undergoing SMT should be consented for risk of stroke or vascular injury from the procedure. A significant increase in neck pain following spinal manipulative therapy warrants immediate medical evaluation. PMID- 12743226 TI - Prevalence and knowledge of transient ischemic attack among US adults. AB - BACKGROUND: Little information is available about public knowledge of TIA and prevalence of a TIA diagnosis. METHODS: The National Stroke Association sponsored a telephone survey by single-stage random-digit dialing of noninstitutionalized US residents > or =18 years old, which was conducted in 1999. Demographic characteristics of participants were compared to the US population to produce weights for projections. Independent predictors of knowledge and diagnosis of TIA were determined by including all demographic characteristics in logistic regression models. RESULTS: Among 10,112 participants, 2.3% reported having been told by a physician that they had a TIA. Older age, lower income, and fewer years of education were independently associated with a diagnosis of TIA. Of those with TIA, only 64% saw a physician within 24 hours of the event. A physician diagnosis of stroke was reported by 2.3% of participants, of whom 19% recalled having had a TIA before the stroke. An additional 3.2% of participants recalled symptoms consistent with TIA but did not seek medical attention. Only 8.2% correctly related the definition of TIA and 8.6% could identify a typical symptom. Men, nonwhites, and those with lower income and fewer years of education were less likely to be knowledgeable about TIA. CONCLUSIONS: An estimated 4.9 million people in the US report a diagnosis of TIA, and many more recall symptoms consistent with TIA but do not seek medical attention. Reducing stroke risk after TIA could have substantial impact on public health but will require public education about the importance of having stroke symptoms evaluated, even if they resolve. PMID- 12743227 TI - Collateral flow and ischemic brain lesions in patients with unilateral carotid artery occlusion. AB - OBJECTIVE: To investigate the association between ischemic brain lesions and intracranial collateral blood flow in patients with unilateral occlusion of the internal carotid artery (ICA). METHODS: Sixty-eight consecutive patients were included. Ischemic lesions on MRI were identified on hard copies, and volume measurements of the lesions were performed on an MR workstation. Intracranial collateral pathways were studied with MR angiography, digital subtraction angiography, and transcranial Doppler sonography. RESULTS: The presence of collateral flow via the anterior communicating artery (ACoA) was associated with a reduction in prevalence (p = 0.01) and volume (p = 0.008) of internal border zone infarcts in the hemisphere ipsilateral to the occluded ICA. Absence of collateral blood flow via the circle of Willis was associated with an increase in prevalence (p = 0.007) and volume (p = 0.005) of internal border zone infarcts. No association between any collateral flow pattern in the circle of Willis and periventricular lesions or lacunar, territorial, or external border zone infarcts was found. No association between collateral flow via the ophthalmic artery or leptomeningeal vessels with any type of ischemic lesion was found. CONCLUSION: Collateral flow via the ACoA is associated with a reduction of the prevalence and volume of internal border zone lesions but not with any other type of ischemic lesion. The presence of a functional posterior communicating artery or secondary collateral pathways is not associated with the prevalence of any type of ischemic lesion. PMID- 12743228 TI - Acute seizures after intracerebral hemorrhage: a factor in progressive midline shift and outcome. AB - OBJECTIVE: To determine whether early seizures that occur frequently after intracerebral hemorrhage (ICH) lead to increased brain edema as manifested by increased midline shift. METHODS: A total of 109 patients with ischemic stroke (n = 46) and intraparenchymal hemorrhage (n = 63) prospectively underwent continuous EEG monitoring after admission. The incidence, timing, and factors associated with seizures were defined. Serial CT brain imaging was conducted at admission, 24 hours, and 48 to 72 hours after hemorrhage and assessed for hemorrhage volume and midline shift. Outcome at time of discharge was assessed using the Glasgow Outcome Scale score. RESULTS: Electrographic seizures occurred in 18 of 63 (28%) patients with ICH, compared with 3 of 46 (6%) patients with ischemic stroke (OR = 5.7, 95% CI 1.4 to 26.5, p < 0.004) during the initial 72 hours after admission. Seizures were most often focal with secondary generalization. Seizures were more common in lobar hemorrhages but occurred in 21% of subcortical hemorrhages. Posthemorrhagic seizures were associated with neurologic worsening on the NIH Stroke Scale (14.8 vs 18.6, p < 0.05) and with an increase in midline shift (+ 2.7 mm vs -2.4 mm, p < 0.03). There was a trend toward increased poor outcome (p < 0.06) in patients with posthemorrhagic seizures. On multivariate analysis, age and initial NIH Stroke Scale score were independent predictors of outcome. CONCLUSION: Seizures occur commonly after ICH and may be nonconvulsive. Seizures are independently associated with increased midline shift after intraparenchymal hemorrhage. PMID- 12743229 TI - Impaired insulin sensitivity among nondiabetic patients with a recent TIA or ischemic stroke. AB - OBJECTIVES: To determine the prevalence of impaired insulin sensitivity among nondiabetic patients with a recent TIA or nondisabling ischemic stroke. METHODS: Eligible subjects were nondiabetic men and women over age 45 years who were hospitalized with a TIA or ischemic stroke. To measure insulin sensitivity, subjects underwent an oral glucose tolerance test between 2 and 6 months after their event. Impaired insulin sensitivity was defined by a value of < or =2.5 on the Composite Insulin Sensitivity Index derived from insulin and glucose values during the test. RESULTS: Between July 2000 and June 2001, we identified 177 eligible patients, among whom 105 declined to participate and 72 enrolled. The median age of participants was 71 years and 46 (64%) were men. The baseline event was stroke for 57 subjects (79%). A history of myocardial infarction (MI) was reported by 14 subjects (19%), and 16 (22%) were obese (body mass index > 30). Fasting glucose was normal (<110 mg/dL) for 58 (80%) participants and impaired (110 to 125 mg/dL) for 14 (20%). Among 72 participants, the median insulin sensitivity index value was 2.6 (range 0.9 to 10.2). The prevalence of impaired insulin sensitivity was 36 of 72 (50%, 95% CI 38% to 62%). Impaired insulin sensitivity was more prevalent among younger patients and patients with obesity, lacunar stroke etiology, and disability (Rankin grade >1). CONCLUSION: Impaired insulin sensitivity is highly prevalent among nondiabetic patients with a recent TIA or nondisabling ischemic stroke. This finding has important therapeutic implications if treatment to improve insulin sensitivity is shown to reduce risk for subsequent stroke and heart disease. PMID- 12743230 TI - Establishment of primary stroke centers: a survey of physician attitudes and hospital resources. AB - OBJECTIVES: To survey US physicians involved in acute stroke care to determine the proportion of hospitals that currently meet the recommended Brain Attack Coalition (BAC) criteria for Primary Stroke Centers (PSC) and obtain opinions regarding the value of stroke centers. METHODS: A survey regarding the BAC guidelines for the establishment of stroke centers was mailed to 3,245 US neurologists, neurosurgeons, and emergency physicians. RESULTS: A total of 1,032 responses were received. Seventy-nine percent (range by specialty 58 to 98%) of respondents believed there was a need for stroke centers. If formal stroke center designation were established, 81% (range 72 to 90%) would like their hospital to become a PSC. Although 77% of respondents believed that their hospital currently met recommended criteria for a PSC, only 7% actually meet all recommended elements. However, 44% of hospitals already provide most acute stroke services. The BAC criteria most frequently lacking were continuing medical education for professional stroke center staff, stroke training for emergency department staff, formal establishment of a stroke unit, and designation of a stroke center director. CONCLUSIONS: The majority of emergency medicine and neuroscience physician respondents involved in acute stroke care support the designation of primary stroke centers. Although respondents globally overestimated the extent to which their facilities currently meet BAC recommended criteria for PSC, detailed responses suggested that over 40% of hospitals possess substantial existing acute stroke care resources and are poised to function as PSC with modest additional administrative and financial commitment. PMID- 12743231 TI - Amyloid beta 1-42 and tau in cerebrospinal fluid after severe traumatic brain injury. AB - OBJECTIVE: To determine whether CSF amyloid beta 1-42 (Abeta-42) and tau have predictive value for prognosis after head injury. METHODS: CSF samples were collected from 29 patients with severe head trauma between 1 and 284 days post trauma. Abeta-42 and tau levels were measured using sandwich ELISA techniques and compared with CSF levels in patients with cognitive disorders and headache. RESULTS: At all time points, concentrations of Abeta-42 were significantly lower in patients with traumatic brain injury (TBI) than in control groups. A significant correlation existed for Abeta-42 levels and outcome of patients. Below a cutoff of 230 pg/mL, the sensitivity of Abeta-42 to discriminate between good outcome (Glasgow Outcome Score 4 and 5) and poor outcome (Glasgow Outcome Score 1 through 3) was 100% at a specificity of 82%. CSF tau levels were significantly higher in patients with TBI than in any control group. In patients with multiple CSF samples collected at various time points between 1 and 32 days after the trauma, tau levels increased early after TBI, peaked in the second week post-trauma, and slowly decreased thereafter. Independent of outcome, all patients had normal tau levels when CSF was collected more than 43 days post trauma. CONCLUSIONS: Abeta-42 and tau may play a potential role in the pathophysiology of TBI. Furthermore, the results of this study suggest that Abeta 42 may be a supportive early predictor for recovery after severe head injury. PMID- 12743232 TI - MRI metrics as surrogate endpoints for EDSS progression in SPMS patients treated with IFN beta-1b. AB - BACKGROUND: Although metrics derived from conventional MRI (cMRI) are widely used as outcome measures in clinical trials of MS, no formal study has been performed to validate cMRI metrics as surrogate endpoints for disability progression in MS. METHODS: A validation procedure was applied to the clinical and MRI data collected in the context of the European randomized, double-blind, placebo controlled trial of interferon beta-1b (IFNbeta-1b) in patients with secondary progressive MS. The Prentice operational criteria were used to assess surrogacy for the number of active lesions seen on the first year T2-weighted MRI scans and the percentage T2 lesion volume change between the baseline and the first year MRI scans. The primary clinical outcome was disability at study exit (3 years), adjusted for the baseline disability. RESULTS: The number of active T2 lesions and the T2 lesion volume percentage change over the first year of the study accounted for 57% of the treatment effect on disability progression over the entire study duration. On the contrary, the same cMRI metrics accounted for 79% of the treatment effect on the relapse rate. CONCLUSIONS: This study shows that the beneficial effect of IFNbeta-1b on disability accumulation in patients with secondary progressive MS is, to a large extent, independent of the changes detected using cMRI. As a consequence, cMRI metrics should not be used as a stand alone measure of outcome in phase III trials of IFNbeta in secondary progressive MS. PMID- 12743233 TI - Inter-rater reliability of a clinical staging of HIV-associated cognitive impairment. AB - OBJECTIVE: To determine the inter-rater reliability of a modification of the Memorial Sloan-Kettering (MSK) Staging for HIV-associated cognitive impairment. METHODS: Data were abstracted on neurologic, neuropsychological, and functional status on 100 individuals participating at four sites in the Northeast AIDS Dementia (NEAD) Consortium cohort study, a longitudinal study of predictors of cognitive impairment in HIV-infected individuals. Neuropsychological performance was defined 1) based on the neuropsychologist's global impression and 2) solely based on neuropsychological test scores. Raters at each site used the abstracted data to assign an MSK stage to each subject blind to any identifying information. Inter-rater reliability was assessed using kappa statistics. Agreement between computer-generated ratings and site-generated ratings was also assessed. RESULTS: Kappa statistics for pair-wise agreement among the sites regarding MSK stage ranged from 0.70-0.91, representing good to excellent agreement between sites. Agreement between computer-generated ratings and site-generated ratings was in the good to excellent range (0.62-0.79). CONCLUSIONS: The authors have modified the MSK rating scale and developed a reliable instrument that can be used in multicenter studies. This instrument will be useful in staging HIV-dementia in future longitudinal studies and will be valuable in increasing accuracy of clinicopathologic studies. PMID- 12743234 TI - Age at focal epilepsy onset varies by sex and hemispheric lateralization. AB - BACKGROUND: Previous studies have shown that interictal epileptiform discharges favor the left hemisphere in adults but the right side in children up until age 5. This may be due to sex-influenced asymmetric brain maturation. To clarify this relationship, the authors analyzed age at epilepsy onset by sex and by lateralization of epileptiform activity. METHODS: An adult epilepsy center long term monitoring database was used to define patients with exclusively unilateral epileptiform findings. Three groups were studied: any epileptiform activity (n = 404), ictal activity (n = 287), and interictal activity (n = 265). The second and third groups were drawn from the first group and the second and third groups overlapped with each other. Side of lateralized finding and sex were analyzed via factorial two-way analysis of variance with the outcome variable being age at epilepsy onset. Comparison analysis included patients with generalized epilepsy (n = 114), nonepileptic seizures (NES, n = 232), and surgical mesial temporal sclerosis (MTS, n = 116). RESULTS: Patients with unilateral epileptiform activity displayed bimodal epilepsy onset ages with infant and adolescent peaks. For patients with a right-sided focus, epilepsy onset was earlier in men (14.4 years) than women (20.7 years). In contrast, among patients with a left-sided focus, epilepsy began earlier in women (18.2 years) than men (19.9 years, p < 0.01). Parallel results were found in unilateral ictal (p < 0.01) and unilateral interictal activity (p = 0.01). Patients with surgical MTS, NES, or generalized seizure showed no similar patterns. CONCLUSIONS: In adult patients with focal epilepsy, sex and lateralized epileptiform abnormalities may be related to age at epilepsy onset. PMID- 12743235 TI - Temporal lobe stimulation reveals anatomic distinction between auditory naming processes. AB - BACKGROUND: Language errors induced by cortical stimulation can provide insight into function(s) supported by the area stimulated. The authors observed that some stimulation-induced errors during auditory description naming were characterized by tip-of-the-tongue responses or paraphasic errors, suggesting expressive difficulty, whereas others were qualitatively different, suggesting receptive difficulty. They hypothesized that these two response types reflected disruption at different stages of auditory verbal processing and that these "subprocesses" might be supported by anatomically distinct cortical areas. OBJECTIVE: To explore the topographic distribution of error types in auditory verbal processing. METHODS: Twenty-one patients requiring left temporal lobe surgery underwent preresection language mapping using direct cortical stimulation. Auditory naming was tested at temporal sites extending from 1 cm from the anterior tip to the parietal operculum. Errors were dichotomized as either "expressive" or "receptive." The topographic distribution of error types was explored. RESULTS: Sites associated with the two error types were topographically distinct from one another. Most receptive sites were located in the middle portion of the superior temporal gyrus (STG), whereas most expressive sites fell outside this region, scattered along lateral temporal and temporoparietal cortex. CONCLUSIONS: Results raise clinical questions regarding the inclusion of the STG in temporal lobe epilepsy surgery and suggest that more detailed cortical mapping might enable better prediction of postoperative language decline. From a theoretical perspective, results carry implications regarding the understanding of structure function relations underlying temporal lobe mediation of auditory language processing. PMID- 12743236 TI - Differential cognitive and behavioral effects of topiramate and valproate. AB - OBJECTIVE: Cognitive effects have been reported during topiramate (TPM) treatment, but effects relative to standard antiepileptic drugs are unclear. METHODS: The authors compared TPM and valproate (VPA) added to carbamazepine (CBZ) in adults with partial seizures. A comprehensive neuropsychological test battery including cognitive, mood, and quality of life measures was used in this multicenter, randomized, double-blind study. After a 4-week baseline, study drug was titrated over 8 weeks to target dosages of 400 mg/d TPM, 2,250 mg/d VPA, or placebo and then maintained for an additional 12 weeks. The neuropsychological test battery was administered at baseline and at the end of titration and maintenance periods. RESULTS: Slightly more patients on TPM dropped out. Neuropsychological data at all three test periods were available for 62 patients. At the end of maintenance, effects of TPM and VPA were comparable, except for two variables (Symbol Digit Modalities Test and Controlled Oral Word Association Test), in which TPM had greater negative effects relative to VPA. The statistical differences appeared to be due in large part to a small subset of patients who were more negatively affected by TPM. Cognitive effects of TPM relative to VPA were greater at the end of titration than at the end of maintenance. CONCLUSIONS: With adjunctive therapy at moderate dose escalation rate, the cognitive effects of TPM are slightly worse overall than VPA in patients who tolerate therapy over several months. PMID- 12743237 TI - Natural history of cognitive decline in the old old. AB - OBJECTIVE: To prospectively examine the occurrence and outcome of cognitive decline in healthy, community-dwelling elders. METHODS: Ninety-five elders (mean age 84 years) who at entry had no cognitive impairment were followed for up to 13 years. Cognitive decline was defined as obtaining either a Clinical Dementia Rating (CDR) = 0.5 or Mini-Mental State Examination (MMSE) score < 24 on two examinations. RESULTS: Three outcomes of aging were determined: intact cognition, persistent cognitive decline without progression to dementia, and dementia. Whereas 49% remained cognitively intact, 51% developed cognitive decline. Mean follow-up to first CDR 0.5 was 3.8 years and age at conversion was 90.0 years. Those who remained cognitively intact had better memory at entry and were less likely to have APOE4 than those who developed cognitive decline. Of the 48 participants with cognitive decline, 27 (56%) developed dementia (CDR > or =1) a mean of 2.8 years later. Participants with cognitive decline who progressed to dementia had poorer confrontation naming at the time of their first CDR 0.5 than those with persistent cognitive decline who did not progress during follow-up. CONCLUSION: The old old are at high risk for developing cognitive decline but many will not progress to dementia in the next 2 to 3 years or even beyond. These findings are important for understanding the prognosis of cognitive decline and for the design of treatment trials for AD. APOE genotype is a risk factor for cognitive decline. PMID- 12743238 TI - Tangle and neuron numbers, but not amyloid load, predict cognitive status in Alzheimer's disease. AB - OBJECTIVE: To examine the relationship between stereologic estimates of AD related pathology and severity of cognitive deficits in brain aging. BACKGROUND: Previous studies reported substantial contributions of neurofibrillary tangles (NFT), amyloid deposits, and neuronal loss to the development of dementia. However, the prediction of cognitive status based on nonstereologic quantification of these measures has led to conflicting results. Such studies have measured densities, rather than absolute numbers, and most do not take into account the potential interaction between the above pathologic hallmarks in a global multivariate analysis. METHODS: Clinicopathologic study in 22 elderly cases. Cognitive status assessed prospectively using the Mini-Mental State Examination (MMSE); stereologic assessment of NFT, unaffected neurons, and total amyloid volume in the CA1 field of the hippocampus, entorhinal cortex, and area 9. Statistical analysis was performed using both univariate and multivariate linear regression models. RESULTS: High total NFT counts but not amyloid volume were strongly associated with a lower number of unaffected neurons in all areas studied. A high proportion of variability in MMSE scores was explained by NFT and neuronal counts in the CA1 field (83% and 85.4%), entorhinal cortex (87.8% and 83.7%), and area 9 (87% and 79%); amyloid volume in the entorhinal cortex, but not in the CA1 field and area 9, accounted for 58.5% of MMSE variability. Multivariate analyses showed that total NFT counts in the entorhinal cortex and area 9 as well as neuron numbers in the CA1 field were the best predictors of MMSE score. CONCLUSIONS: These new stereologic data indicate that neuronal pathology in hippocampal formation and frontal cortex closely reflects the progression of cognitive deficits in brain aging and AD. They also demonstrate that amyloid volume has no additional predictive value, in terms of clinicopathologic correlations, beyond its interaction with NFT. PMID- 12743239 TI - Altered central somatosensory processing in chronic pain patients with "hysterical" anesthesia. AB - OBJECTIVE: The authors hypothesized that central factors may underlie sensory deficits in patients with nondermatomal somatosensory deficits (NDSD) and that functional brain imaging would reveal altered responses in supraspinal nuclei. BACKGROUND: Patients with chronic pain frequently present with NDSD, ranging from hypoesthesia to complete anesthesia in the absence of substantial pathology and often in association with motor weakness and occasional paralysis. Patients with pain and such pseudoneurologic symptoms can be classified as having both a pain disorder and a conversion disorder (Diagnostic and Statistical Manual of Mental Disorders-IV classification). METHODS: The authors tested their hypothesis with functional MRI (fMRI) of brush and noxious stimulation-evoked brain responses in four patients with chronic pain and NDSD. RESULTS: The fMRI findings revealed altered somatosensory-evoked responses in specific forebrain areas. Unperceived stimuli failed to activate areas that were activated with perceived touch and pain: notably, the thalamus, posterior region of the anterior cingulate cortex (ACC), and Brodmann area 44/45. Furthermore, unperceived stimuli were associated with deactivations in primary and secondary somatosensory cortex (S1, S2), posterior parietal cortex, and prefrontal cortex. Finally, unperceived (but not perceived) stimuli activated the rostral ACC. CONCLUSIONS: Diminished perception of innocuous and noxious stimuli is associated with altered activity in many parts of the somatosensory pathway or other supraspinal areas. The cortical findings indicate a neurobiological component for at least part of the symptoms in patients presenting with nondermatomal somatosensory deficits. PMID- 12743240 TI - Lamotrigine for HIV-associated painful sensory neuropathies: a placebo-controlled trial. AB - OBJECTIVE: To evaluate the efficacy and tolerability of lamotrigine (LTG) for the treatment of pain in HIV-associated sensory neuropathies. METHODS: In a randomized, double-blind study, patients with HIV-associated distal sensory polyneuropathy (DSP) received LTG or placebo during a 7-week dose escalation phase followed by a 4-week maintenance phase. Randomization was stratified according to whether or not patients were currently using neurotoxic antiretroviral therapy (ART). RESULTS: The number of patients randomized was 92 (62 LTG, 30 placebo) in the stratum receiving neurotoxic ART and 135 (88 LTG, 47 placebo) in the stratum not receiving neurotoxic ART. Mean change from baseline in Gracely Pain Scale score for average pain was not different between LTG and placebo at the end of the maintenance phase in either stratum, but the slope of the change in Gracely Pain Scale score for average pain reflected greater improvement with LTG than with placebo in the stratum receiving neurotoxic ART (p = 0.004), as did the mean change from baseline scores on the Visual Analogue Scale for Pain Intensity and the McGill Pain Assessment Scale and patient and clinician ratings of global impression of change in pain (p VP2 + VP4) did not occur, and the unassembled mutant capsid subunits were degraded with a half-life of 15 min. Drug was not required by mutant virus for attachment, uncoating, RNA synthesis and protein synthesis, and polyprotein processing except maturation cleavage. The requirement of drug for assembly of mutant pentamers to form provirions and the rapid assembly of preformed subunits (synthesized in the absence of drug) after drug addition suggested that after native pentamers (P5) have been formed they must be converted to an assembly active state (P5(*)), possibly by a conformational change induced by the binding of drug. We propose that pocket factor plays the same role in wild-type virus. In addition, we also report the construction and the properties of a full-length cDNA clone of HRV16, pR16.11, which produces in vitro transcripts with infectivity similar to that of virion RNA. This cDNA clone is available at the American Type Culture Collection. PMID- 12743279 TI - CpG methylation of human papillomavirus type 16 DNA in cervical cancer cell lines and in clinical specimens: genomic hypomethylation correlates with carcinogenic progression. AB - Infection with genital human papillomaviruses (HPVs) is the primary cause of cervical cancer. The infection is widespread, and little is known about the secondary factors associated with progression from subclinical infection to invasive carcinoma. Here we report that HPV genomes are efficiently targeted in vivo by CpG methylation, a well-known mechanism of transcriptional repression. Indeed, it has been shown previously that in vitro-methylated HPV type 16 (HPV 16) DNA is transcriptionally repressed after transfection into cell cultures. By using a scan with the restriction enzyme McrBC, we observed a conserved profile of CpG hyper- and hypomethylation throughout the HPV-16 genomes of the tumor derived cell lines SiHa and CaSki. Methylation is particularly high in genomic segments overlying the late genes, while the long control region (LCR) and the oncogenes are unmethylated in the single HPV-16 copy in SiHa cells. In 81 patients from two different cohorts, the LCR and the E6 gene of HPV-16 DNA were found to be hypermethylated in 52% of asymptomatic smears, 21.7% of precursor lesions, and 6.1% of invasive carcinomas. This suggests that neoplastic transformation may be suppressed by CpG methylation, while demethylation occurs as the cause of or concomitant with neoplastic progression. These prevalences of hyper- and hypomethylation also indicate that CpG methylation plays an important role in the papillomavirus life cycle, which takes place in asymptomatic infections and precursor lesions but not in carcinomas. Bisulfite modification revealed that in most of the HPV-16 genomes of CaSki cells and of asymptomatic patients, all 11 CpG dinucleotides that overlap with the enhancer and the promoter were methylated, while in SiHa cells and cervical lesions, the same 11 or a subset of CpGs remained unmethylated. Our report introduces papillomaviruses as models to study the mechanism of CpG methylation, opens research on the importance of this mechanism during the viral life cycle, and provides a marker relevant for the etiology and diagnosis of cervical cancer. PMID- 12743281 TI - Resolution of parvovirus dimer junctions proceeds through a novel heterocruciform intermediate. AB - The minute virus of mice initiator protein, NS1, excises new copies of the left viral telomere in a single sequence orientation, dubbed flip, during resolution of the junction between monomer genomes in palindromic dimer intermediate duplexes. We examined this reaction in vitro using both (32)P-end-labeled linear substrates and similar unlabeled templates labeled by incorporation of [alpha (32)P]TTP during the synthesis. The observed products suggest a resolution model that explains conservation of the hairpin sequence and in which a novel heterocruciform intermediate plays a crucial role. In vitro, NS1 initiates two replication pathways from OriL(TC), the single active origin embedded in one arm of the dimer junction. NS1-mediated nicking liberates a base-paired 3' nucleotide to prime DNA synthesis and, in a reaction we call "read-through synthesis," forks established while the substrate is a linear duplex synthesize DNA in the flop orientation, leading to DNA amplification but not to junction resolution. Nicking leaves NS1 covalently attached to the 5' end of the DNA, where it can serve as a 3'-to-5' helicase, unwinding the NS1-associated strand. In the second pathway, resolution substrates are created when such unwinding induces the palindrome to reconfigure into a cruciform prior to fork assembly. New forks can then synthesize DNA in the flip orientation, copying one cruciform arm and creating a heterocruciform intermediate. Resolution proceeds via hairpin transfer in the extended arm of the heterocruciform, which releases one covalently closed duplex telomere and a partially single-stranded junction intermediate. We suggest that the latter intermediate is finally resolved via an NS1-induced single-strand nick at the otherwise inactive origin, OriL(GAA). PMID- 12743282 TI - Binding of CCAAT displacement protein CDP to adenovirus packaging sequences. AB - Adenovirus (Ad) type 5 DNA packaging is initiated in a polar fashion from the left end of the genome. The packaging process is dependent upon the cis-acting packaging domain located between nucleotides 194 and 380. Seven A/T-rich repeats have been identified within this domain that direct packaging. A1, A2, A5, and A6 are the most important repeats functionally and share a bipartite sequence motif. Several lines of evidence suggest that there is a limiting trans-acting factor(s) that plays a role in packaging. Two cellular activities that bind to minimal packaging domains in vitro have been previously identified. These binding activities are P complex, an uncharacterized protein(s), and chicken ovalbumin upstream promoter transcription factor (COUP-TF). In this work, we report that a third cellular protein, octamer-1 protein (Oct-1), binds to minimal packaging domains. In vitro binding analyses and in vivo packaging assays were used to examine the relevance of these DNA binding activities to Ad DNA packaging. The results of these experiments reveal that COUP-TF and Oct-1 binding does not play a functional role in Ad packaging, whereas P-complex binding directly correlates with packaging function. We demonstrate that P complex contains the cellular protein CCAAT displacement protein (CDP) and that full-length CDP is found in purified virus particles. In addition to cellular factors, previous evidence indicates that viral factors play a role in the initiation of viral DNA packaging. We propose that CDP, in conjunction with one or more viral proteins, binds to the packaging sequences of Ad to initiate the encapsidation process. PMID- 12743283 TI - Palmitoylation of the Autographa californica multicapsid nucleopolyhedrovirus envelope glycoprotein GP64: mapping, functional studies, and lipid rafts. AB - Budded virions (BV) of the baculovirus Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) contain a major envelope glycoprotein known as GP64, which was previously shown to be palmitoylated. In the present study, we used truncation and amino acid substitution mutations to map the palmitoylation site to cysteine residue 503. Palmitoylation of GP64 was not detected when Cys503 was replaced with alanine or serine. Palmitoylation-minus forms of GP64 were used to replace wild-type GP64 in AcMNPV, and these viruses were used to examine potential functions of GP64 palmitoylation in the context of the infection cycle. Analysis by immunoprecipitation and cell surface studies revealed that palmitoylation of GP64 did not affect GP64 synthesis or its transport to the cell surface in Sf9 cells. GP64 proteins lacking palmitoylation also mediated low-pH triggered membrane fusion in a manner indistinguishable from that of wild-type GP64. Cells infected with viruses expressing palmitoylation-minus forms of GP64 produced infectious virions at levels similar to those from cells infected with wild-type AcMNPV. In combination, these data suggest that virus entry and exit in Sf9 cells were not significantly affected by GP64 palmitoylation. To determine whether GP64 palmitoylation affected the association of GP64 with membrane microdomains, the potential association of GP64 with lipid raft microdomains was examined. These experiments showed that: (i) AcMNPV-infected Sf9 cell membranes contain lipid raft microdomains, (ii) GP64 association with lipid rafts was not detected in infected Sf9 cells, and (iii) GP64 palmitoylation did not affect the apparent exclusion of GP64 from lipid raft microdomains. PMID- 12743284 TI - Hepatitis B virus X protein and simian virus 5 V protein exhibit similar UV-DDB1 binding properties to mediate distinct activities. AB - The UV-damaged DNA-binding activity protein (UV-DDB) consists of two subunits, DDB1 and DDB2, and functions in DNA repair and cell cycle regulation. The DDB1 subunit is a target for the hepatitis B virus X protein (HBx). Binding of HBx to DDB1 interferes with cell growth and viability in culture and has been implicated in the establishment of viral infection. DDB1 also interacts with the V proteins encoded by several paramyxoviruses including simian virus 5 (SV5), which prevent interferon signaling by targeting either STAT1 or STAT2 proteins for proteolysis. The role of V binding to DDB1, however, remains unclear. Here we show that the V protein of SV5 (SV5-V) and HBx exhibit strikingly similar DDB1 binding properties. Thus, SV5-V and HBx bind to DDB1 in a mutually exclusive manner, and SV5-V shares with HBx the ability to enhance the steady-state levels of DDB1 and to inhibit its association with DDB2. Yet only HBx induces cell death, and SV5-V can prevent HBx from doing so by blocking its interaction with DDB1. Binding of SV5-V to DDB1 may serve another function, since SV5-V shows a decreased ability to induce STAT1 degradation in cells expressing reduced amounts of DDB1. These findings demonstrate that HBx performs a unique function through its association with DDB1 for which SV5-V cannot substitute and suggest that SV5-V and HBx have evolved to bind DDB1 to achieve distinct functions, both by a mechanism that does not involve DDB2. PMID- 12743285 TI - The simian immunodeficiency virus 5' untranslated leader sequence plays a role in intracellular viral protein accumulation and in RNA packaging. AB - We investigated the role of 5' untranslated leader sequences of simian immunodeficiency virus (SIV(mac239)) in RNA encapsidation and protein expression. A series of progressively longer deletion mutants was constructed with a common endpoint six nucleotides upstream of the gag initiation codon and another endpoint at the 3' end of the primer binding site (PBS). We found that efficient intracellular Gag-Pol protein accumulation required the region between the PBS and splice donor (SD) site. Marked reduction of genomic RNA packaging was observed with all the deletion mutants that involved sequences at both the 5' and at the 3' ends of the major SD site, and increased nonspecific RNA incorporation could be detected in these mutants. RNA encapsidation was affected only modestly by a deletion of 54 nucleotides at the 3' end of the SD site when the mutant construct pDelta54 was transfected alone. In contrast, the amount of pDelta54 genomic RNA incorporated into particles was reduced more than 10-fold when this mutant was cotransfected with a construct specifying an RNA molecule with a wild type packaging signal. Therefore, we conclude that the 175 nucleotides located 5' of the gag initiation codon are critical for efficient and selective incorporation of genomic RNA into virions. This location of the SIV Psi element provides the means for efficient discrimination between viral genomic and spliced RNAs. PMID- 12743286 TI - Transport of the intracisternal A-type particle Gag polyprotein to the endoplasmic reticulum is mediated by the signal recognition particle. AB - Intracisternal A-type particles (IAP) are defective endogenous retroviruses that accumulate in the endoplasmic reticulum (ER) of rodent cells. The enveloped particles are produced by assembly and budding of IAP Gag polyproteins at the ER membrane. In this study, we analyzed the specific ER transport of the Gag polyprotein of the IAP element MIA14. To this end, we performed in vitro translation of Gag in the presence of microsomal membranes or synthetic proteoliposomes followed by membrane sedimentation or flotation. ER binding of IAP Gag occurred mostly cotranslationally, and Gag polyproteins interacted specifically with proteoliposomes containing only signal recognition particle (SRP) receptor and the Sec61p complex, which form the minimal ER translocation apparatus. The direct participation of SRP in ER targeting of IAP Gag was demonstrated in cross-linking and immunoprecipitation experiments. The IAP polyprotein was not translocated into the ER; it was found to be tightly associated with the cytoplasmic side of the ER membrane but did not behave as an integral membrane protein. Substituting the functional signal peptide of preprolactin for the hydrophobic sequence at the N terminus of IAP Gag also did not result in translocation of the chimeric protein into the ER lumen, and grafting the IAP hydrophobic sequence onto preprolactin failed to yield luminal transport as well. These results suggest that the N-terminal hydrophobic region of the IAP Gag polyprotein functions as a transport signal which mediates SRP dependent ER targeting, but polyprotein translocation or integration into the membrane is prevented by the signal sequence itself and by additional regions of Gag. PMID- 12743287 TI - Comparative immunogenicity in rhesus monkeys of DNA plasmid, recombinant vaccinia virus, and replication-defective adenovirus vectors expressing a human immunodeficiency virus type 1 gag gene. AB - Cellular immune responses, particularly those associated with CD3(+) CD8(+) cytotoxic T lymphocytes (CTL), play a primary role in controlling viral infection, including persistent infection with human immunodeficiency virus type 1 (HIV-1). Accordingly, recent HIV-1 vaccine research efforts have focused on establishing the optimal means of eliciting such antiviral CTL immune responses. We evaluated several DNA vaccine formulations, a modified vaccinia virus Ankara vector, and a replication-defective adenovirus serotype 5 (Ad5) vector, each expressing the same codon-optimized HIV-1 gag gene for immunogenicity in rhesus monkeys. The DNA vaccines were formulated with and without one of two chemical adjuvants (aluminum phosphate and CRL1005). The Ad5-gag vector was the most effective in eliciting anti-Gag CTL. The vaccine produced both CD4(+) and CD8(+) T-cell responses, with the latter consistently being the dominant component. To determine the effect of existing antiadenovirus immunity on Ad5-gag-induced immune responses, monkeys were exposed to adenovirus subtype 5 that did not encode antigen prior to immunization with Ad5-gag. The resulting anti-Gag T-cell responses were attenuated but not abolished. Regimens that involved priming with different DNA vaccine formulations followed by boosting with the adenovirus vector were also compared. Of the formulations tested, the DNA-CRL1005 vaccine primed T-cell responses most effectively and provided the best overall immune responses after boosting with Ad5-gag. These results are suggestive of an immunization strategy for humans that are centered on use of the adenovirus vector and in which existing adenovirus immunity may be overcome by combined immunization with adjuvanted DNA and adenovirus vector boosting. PMID- 12743288 TI - The unique vertex of bacterial virus PRD1 is connected to the viral internal membrane. AB - Icosahedral double-stranded DNA (dsDNA) bacterial viruses are known to package their genomes into preformed procapsids via a unique portal vertex. Bacteriophage PRD1 differs from the more commonly known icosahedral dsDNA phages in that it contains an internal lipid membrane. The packaging of PRD1 is known to proceed via preformed empty capsids. Now, a unique vertex has been shown to exist in PRD1. We show in this study that this unique vertex extends to the virus internal membrane via two integral membrane proteins, P20 and P22. These small membrane proteins are necessary for the binding of the putative packaging ATPase P9, via another capsid protein, P6, to the virus particle. PMID- 12743289 TI - Infection with Theiler's murine encephalomyelitis virus directly induces proinflammatory cytokines in primary astrocytes via NF-kappaB activation: potential role for the initiation of demyelinating disease. AB - Theiler's virus infection in the central nervous system (CNS) induces a demyelinating disease very similar to human multiple sclerosis. We have assessed cytokine gene activation upon Theiler's murine encephalomyelitis virus (TMEV) infection and potential mechanisms in order to delineate the early events in viral infection that lead to immune-mediated demyelinating disease. Infection of SJL/J primary astrocyte cultures induces selective proinflammatory cytokine genes (interleukin-12p40 [IL-12p40], IL-1, IL-6, tumor necrosis factor alpha, and beta interferon [IFN-beta]) important in the innate immune response to infection. We find that TMEV-induced cytokine gene expression is mediated by the NF-kappaB pathway based on the early nuclear NF-kappaB translocation and suppression of cytokine activation in the presence of specific inhibitors of the NF-kappaB pathway. Further studies show this to be partly independent of dsRNA-dependent protein kinase (PKR) and IFN-alpha/beta pathways. Altogether, these results demonstrate that infection of astrocytes and other CNS-resident cells by TMEV provides the early NF-kappaB-mediated signals that directly activate various proinflammatory cytokine genes involved in the initiation and amplification of inflammatory responses in the CNS known to be critical for the development of immune-mediated demyelination. PMID- 12743290 TI - Structure of intracellular mature vaccinia virus visualized by in situ atomic force microscopy. AB - Vaccinia virus, the basis of the smallpox vaccine, is one of the largest viruses to replicate in humans. We have used in situ atomic force microscopy (AFM) to directly visualize fully hydrated, intact intracellular mature vaccinia virus (IMV) virions and chemical and enzymatic treatment products thereof. The latter included virion cores, core-enveloping coats, and core substructures. The isolated coats appeared to be composed of a highly cross-linked protein array. AFM imaging of core substructures indicated association of the linear viral DNA genome with a segmented protein sheath forming an extended approximately 16-nm diameter filament with helical surface topography; enclosure of this filament within a 30- to 40-nm-diameter tubule which also shows helical topography; and enclosure of the folded, condensed 30- to 40-nm-diameter tubule within the core by a wall covered with peg-like projections. Proteins observed attached to the 30 to 40-nm-diameter tubules may mediate folding and/or compaction of the tubules and/or represent vestiges of the core wall and/or pegs. An accessory "satellite domain" was observed protruding from the intact core. This corresponded in size to isolated 70- to 100-nm-diameter particles that were imaged independently and might represent detached accessory domains. AFM imaging of intact virions indicated that IMV underwent a reversible shrinkage upon dehydration (as much as 2.2- to 2.5-fold in the height dimension), accompanied by topological and topographical changes, including protrusion of the satellite domain. As shown here, the chemical and enzymatic dissection of large, asymmetrical virus particles in combination with in situ AFM provides an informative complement to other structure determination techniques. PMID- 12743291 TI - Transformation of rodent fibroblasts by the jaagsiekte sheep retrovirus envelope is receptor independent and does not require the surface domain. AB - Jaagsiekte sheep retrovirus (JSRV) is the etiological agent of a contagious lung cancer of sheep known as ovine pulmonary adenocarcinoma (OPA). Expression of the JSRV envelope protein (Env) is sufficient to transform immortalized and primary fibroblasts, but the precise mechanisms of this process are not known. The cellular receptor for JSRV is hyaluronidase 2 (Hyal-2), the product of a putative tumor suppressor gene that in humans maps to a chromosomal region frequently deleted in the development of lung and breast cancers. Here we report studies to determine whether the Hyal-2-JSRV Env interaction plays a role in virus-induced transformation of rodent fibroblasts. Chimeric Env proteins between JSRV and the unrelated murine retroviruses Moloney murine leukemia virus (MMuLV) and mouse mammary tumor virus (MMTV) showed cell surface expression comparable to that of wild-type MMuLV Env and rescued infection of MMuLV particle pseudotypes. Interestingly, an MMuLV-JSRV chimera in which the putative receptor binding domain (RBD) and proline-rich region (PRR) of JSRV Env were replaced by the RBD and PRR of MMuLV induced transformation of 208F, a rodent fibroblast line. Cell lines derived from foci of MMuLV-JSRV chimera-transformed 208F cells grew in soft agar and showed Akt activation, a hallmark of JSRV-transformed rodent fibroblasts. Transformation assays performed using proteins with amino-terminal deletion mutations showed that the carboxy-terminal 141 amino acids of the transmembrane subunit (TM) were sufficient to induce cell transformation when targeted to the membrane with a myristoylation signal. Thus, the JSRV TM is necessary and sufficient to transform rodent fibroblasts. Taken together these results indicate that the interaction with Hyal-2 at least is not an essential determinant of JSRV-induced transformation of fibroblasts and that the viral TM functions essentially as an oncoprotein. PMID- 12743292 TI - Herpes simplex virus type 1 portal protein UL6 interacts with the putative terminase subunits UL15 and UL28. AB - The herpes simplex virus type 1 (HSV-1) UL6, UL15, and UL28 proteins are essential for cleavage of replicated concatemeric viral DNA into unit length genomes and their packaging into a preformed icosahedral capsid known as the procapsid. The capsid-associated UL6 DNA-packaging protein is located at a single vertex and is thought to form the portal through which the genome enters the procapsid. The UL15 protein interacts with the UL28 protein, and both are strong candidates for subunits of the viral terminase, a key component of the molecular motor that drives the DNA into the capsid. To investigate the association of the UL6 protein with the UL15 and UL28 proteins, the three proteins were produced in large amounts in insect cells with the baculovirus expression system. Interactions between UL6 and UL28 and between UL6 and UL15 were identified by an immunoprecipitation assay. These results were confirmed by transiently expressing wild-type and mutant proteins in mammalian cells and monitoring their distribution by immunofluorescence. In cells expressing the single proteins, UL6 and UL15 were concentrated in the nuclei whereas UL28 was found in the cytoplasm. When the UL6 and UL28 proteins were coexpressed, UL28 was redistributed to the nuclei, where it colocalized with UL6. In cells producing either of two cytoplasmic UL6 mutant proteins and a functional epitope-tagged form of UL15, the UL15 protein was concentrated with the mutant UL6 protein in the cytoplasm. These observed interactions of UL6 with UL15 and UL28 are likely to be of major importance in establishing a functional DNA-packaging complex at the portal vertex of the HSV-1 capsid. PMID- 12743294 TI - Gene expression profiling of the cellular transcriptional network regulated by alpha/beta interferon and its partial attenuation by the hepatitis C virus nonstructural 5A protein. AB - Alpha/beta interferons (IFN-alpha/beta) induce potent antiviral and antiproliferative responses and are used to treat a wide range of human diseases, including chronic hepatitis C virus (HCV) infection. However, for reasons that remain poorly understood, many HCV isolates are resistant to IFN therapy. To better understand the nature of the cellular IFN response, we examined the effects of IFN treatment on global gene expression by using several types of human cells, including HeLa cells, liver cell lines, and primary fetal hepatocytes. In response to IFN, 50 of the approximately 4,600 genes examined were consistently induced in each of these cell types and another 60 were induced in a cell type-specific manner. A search for IFN-stimulated response elements (ISREs) in genomic DNA located upstream of IFN-stimulated genes revealed both previously identified and novel putative ISREs. To determine whether HCV can alter IFN-regulated gene expression, we performed microarray analyses on IFN treated HeLa cells expressing the HCV nonstructural 5A (NS5A) protein and on IFN treated Huh7 cells containing an HCV subgenomic replicon. NS5A partially blocked the IFN-mediated induction of 14 IFN-stimulated genes, an effect that may play a role in HCV resistance to IFN. This block may occur through repression of ISRE mediated transcription, since NS5A also inhibited the IFN-mediated induction of a reporter gene driven from an ISRE-containing promoter. In contrast, the HCV replicon had very little effect on IFN-regulated gene expression. These differences highlight the importance of comparing results from multiple model systems when investigating complex phenomena such as the cellular response to IFN and viral mechanisms of IFN resistance. PMID- 12743293 TI - U.S. Human immunodeficiency virus type 1 epidemic: date of origin, population history, and characterization of early strains. AB - Human immunodeficiency virus (HIV) type 1 subtype B sequences (whole envelope and the p17 region of gag) were obtained from peripheral blood mononuclear cell samples collected in 1981 from seven HIV-infected U.S. individuals and in 1982 from one infected Canadian resident. Phylogenetic and nucleotide distance analyses were performed by using database sequences representing North American strains collected from 1978 to 1995. The estimated phylogeny was starlike, with early strains represented on different lineages. When sequences were grouped by years of collection, nucleotide distance comparisons demonstrated an increase in diversity over time and indicated that contemporary strains are more closely related to early epidemic strains than to each other. Using a recently developed likelihood ratio reduction procedure, the date of origin of the U.S. epidemic was estimated to be 1968 +/- 1.4 years. A coalescent approach was also used to estimate the population history of the U.S. subtype B epidemic. Our analyses provide new information that implies an exponential growth rate from the beginning of the U.S. HIV epidemic. The dating results suggest a U.S. introduction date (or date of divergence from the most recent common ancestor) that precedes the date of the earliest known AIDS cases in the late 1970s. Furthermore, the estimated epidemic growth curve shows a period of exponential growth that preceded most of the early documented cases and also indicates a leveling of prevalence rates in the recent past. PMID- 12743295 TI - Intracellular targeting of Gag proteins of the Drosophila telomeric retrotransposons. AB - Drosophila has two non-long-terminal-repeat (non-LTR) retrotransposons that are unique because they have a defined role in chromosome maintenance. These elements, HeT-A and TART, extend chromosome ends by successive transpositions, producing long arrays of head-to-tail repeat sequences. These arrays appear to be analogous to the arrays produced by telomerase on chromosomes of other organisms. While other non-LTR retrotransposons transpose to many chromosomal sites, HeT-A and TART transpose only to chromosome ends. Although HeT-A and TART belong to different subfamilies of non-LTR retrotransposons, they encode very similar Gag proteins, which suggests that Gag proteins are involved in their unique transposition targeting. We have recently shown that both Gags localize efficiently to nuclei where HeT-A Gag forms structures associated with telomeres. TART Gag does not associate with telomeres unless HeT-A Gag is present, suggesting a symbiotic relationship in which HeT-A Gag provides telomeric targeting. We now report studies to identify amino acid regions responsible for different aspects of the intracellular targeting of these proteins. Green fluorescent protein-tagged deletion derivatives were expressed in cultured Drosophila cells. The intracellular localization of these proteins shows the following. (i) Several regions that direct subcellular localizations or cluster formation are found in both Gags and are located in equivalent regions of the two proteins. (ii) Regions important for telomere association are present only in HeT A Gag. These are present at several places in the protein, are not redundant, and cannot be complemented in trans. (iii) Regions containing zinc knuckle and major homology region motifs, characteristic of retroviral Gags, are involved in protein-protein interactions of the telomeric Gags, as they are in retroviral Gags. PMID- 12743296 TI - STAT3 ubiquitylation and degradation by mumps virus suppress cytokine and oncogene signaling. AB - Mumps virus is a common infectious agent of humans, causing parotitis, meningitis, encephalitis, and orchitis. Like other paramyxoviruses in the genus Rubulavirus, mumps virus catalyzes the proteasomal degradation of cellular STAT1 protein, a means for escaping antiviral responses initiated by alpha/beta and gamma interferons. We demonstrate that mumps virus also eliminates cellular STAT3, a protein that mediates transcriptional responses to cytokines, growth factors, nonreceptor tyrosine kinases, and a variety of oncogenic stimuli. STAT1 and STAT3 are independently targeted by a single mumps virus protein, called V, that assembles STAT-directed ubiquitylation complexes from cellular components, including STAT1, STAT2, STAT3, DDB1, and Cullin4A. Consequently, mumps virus V protein prevents responses to interleukin-6 and v-Src signals and can induce apoptosis in STAT3-dependent multiple myeloma cells and transformed murine fibroblasts. These findings demonstrate a unique cytokine and oncogene evasion property of mumps virus that provides a molecular basis for its observed oncolytic properties. PMID- 12743297 TI - Enhancement of recombinant adeno-associated virus type 2-mediated transgene expression in a lung epithelial cell line by inhibition of the epidermal growth factor receptor. AB - Recombinant adeno-associated viruses (rAAVs) have attracted considerable interest as gene delivery systems because they show long-term expression in vivo and transduce numerous cell types. Limitations to successful gene transduction from rAAVs have prompted investigations of a variety of treatments to enhance transgene expression from rAAV vectors. Tyrphostin-1, an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, dramatically enhances rAAV transgene expression. Elegant studies have demonstrated that a single-strand D-sequence binding protein (ssDBP) is phosphorylated by EGFR and binds to the D sequence element in the AAV terminal repeat (TR). Binding of the Tyr-phosphorylated ssDBP prevents conversion of single-stranded vector DNA to a double-strand conformation. We observed dramatic increases in transgene expression in lung epithelial cells (IB3) with tyrphostin treatment. Gel shift analysis of ssDBP revealed that its DNA binding characteristics were unchanged after tyrphostin treatment or adenovirus infection. Tyrphostin stimulated rAAV transgene expression to a greater extent than adenovirus coinfection. Southern hybridizations revealed that the vector DNA remained in the single-strand conformation in tyrphostin-treated cells but double-stranded replicative form monomer DNA was most abundant in adenovirus-infected cells. Northern analyses revealed that tyrphostin treatment enhanced mRNA accumulation more than in adenovirus-infected cultures even though replicative form DNA was undetectable. Analysis of the JNK, ERK, and p38K mitogen-activated protein kinase pathways revealed that tyrphostin treatment stimulated the activity of JNK and p38K. Our data suggest that tyrphostin-induced alteration of stress response pathways results in dramatic enhancement of transcription on linear vector DNA templates in the IB3 cell line. These results expand the downstream targets of the EGFR in regulating rAAV transduction. PMID- 12743298 TI - Unique pattern of convergent envelope evolution in simian immunodeficiency virus infected rapid progressor macaques: association with CD4-independent usage of CCR5. AB - The rate of disease development in simian immunodeficiency virus (SIV) infection of macaques varies considerably among individual macaques. While the majority of macaques inoculated with pathogenic SIV develop AIDS within a period of 1 to 2 years, a minority exhibit a rapid disease course characterized by absence or transience of humoral and cellular immune responses and high levels of virus replication with widespread dissemination of SIV in macrophages and multinucleated giant cells. The goal of this study was to examine viral evolution in three SIVsmE543-3-inoculated rapid progressors to determine the contribution of viral evolution to the development of rapid disease and the effect of the absence of immune pressure upon viral evolution. PCR was used to amplify and clone the entire SIV genome from tissues collected at necropsy, and the course of viral evolution was assessed by env sequences cloned from sequential plasma samples of one rapid progressor (RP) macaque. The majority of sequence changes in RP macaques occurred in the envelope gene. Substitutions were observed in all three animals at specific conserved residues in envelope, including loss of a glycosylation site in V1/V2, a D-to-N/V substitution in a highly conserved GDPE motif, and a P-to-V/H/T substitution in the V3 loop analog. A cell-cell fusion assay revealed that representative env clones utilized CCR5 as a coreceptor, independent of CD4. The selection of specific substitutions in envelope in RP macaques suggests novel selection pressures on virus in such animals and suggests that viral variants that evolve in these animals may play a role in disease progression. PMID- 12743299 TI - A new Sendai virus vector deficient in the matrix gene does not form virus particles and shows extensive cell-to-cell spreading. AB - A new recombinant Sendai virus vector (SeV/DeltaM), in which the gene encoding matrix (M) protein was deleted, was recovered from cDNA and propagated in a packaging cell line expressing M protein by using a Cre/loxP induction system. The titer of SeV/DeltaM carrying the enhanced green fluorescent protein gene in place of the M gene was 7 x 10(7) cell infectious units/ml or more. The new vector showed high levels of infectivity and gene expression, similar to those of wild-type SeV vector, in vitro and in vivo. Virus maturation into a particle was almost completely abolished in cells infected with SeV/DeltaM. Instead, SeV/DeltaM infection brought about a significant increase of syncytium formation under conditions in which the fusion protein was proteolytically cleaved and activated by trypsin-like protease. This shows that SeV/DeltaM spreads markedly to neighboring cells in a cell-to-cell manner, because both hemagglutinin neuraminidase and active fusion proteins are present at very high levels on the surface of cells infected with SeV/DeltaM. Thus, SeV/DeltaM is a novel type of vector with the characteristic features of loss of virus particle formation and gain of cell-to-cell spreading via a mechanism dependent on the activation of the fusion protein. PMID- 12743300 TI - Clearance of a productive lentivirus infection in calves experimentally inoculated with caprine arthritis-encephalitis virus. AB - Lentiviruses have long been considered host-specific pathogens, but several recent observations demonstrated their capacity to conquer new hosts from different species, genera, and families. From these cross-species infections emerged new animal and human infectious diseases. The successful colonization and adaptation of a lentivirus to a nonnatural host depends on unspecific and specific host barriers. Some of those barriers exert a relative control of viral replication (i.e., cytotoxic T-lymphocyte response, viral inhibitory factors), but none of them was found able to totally clear the infection once the retrovirus is fully adapted in its host. In this study we examined the evolution of the host-lentivirus interactions occurring in an experimental animal model of cross-species infection in order to analyze the efficiency of those barriers in preventing the establishment of a persistent infection. Five newborn calves were inoculated with caprine arthritis-encephalitis virus (CAEV), and the evolution of infection was studied for more than 12 months. All the animals seroconverted in the first 0.75 to 1 month following the inoculation and remained seropositive for the remaining time of the experiment. Viral infection was productive during 4 months with isolation of replication competent virus from the blood cells and organs of the early euthanized animals. After 4 months of infection, neither replication-competent virus nor virus genome could be detected in blood cells or in the classical target organs, even after an experimental immunosuppression. No evidence of in vitro restriction of CAEV replication was observed in cells from tissues explanted from organs of these calves. These data provide the demonstration of a natural clearance of lentivirus infection following experimental inoculation of a nonnatural host, enabling perspectives of development of new potential vaccine strategies to fight against lentivirus infections. PMID- 12743301 TI - The oligomerization domain of VP3, the scaffolding protein of infectious bursal disease virus, plays a critical role in capsid assembly. AB - Infectious bursal disease virus (IBDV) capsids are formed by a single protein layer containing three polypeptides, pVP2, VP2, and VP3. Here, we show that the VP3 protein synthesized in insect cells, either after expression of the complete polyprotein or from a VP3 gene construct, is proteolytically degraded, leading to the accumulation of product lacking the 13 C-terminal residues. This finding led to identification of the VP3 oligomerization domain within a 24-amino-acid stretch near the C-terminal end of the polypeptide, partially overlapping the VP1 binding domain. Inactivation of the VP3 oligomerization domain, by either proteolysis or deletion of the polyprotein gene, abolishes viruslike particle formation. Formation of VP3-VP1 complexes in cells infected with a dual recombinant baculovirus simultaneously expressing the polyprotein and VP1 prevented VP3 proteolysis and led to efficient virus-like particle formation in insect cells. PMID- 12743302 TI - Development and application of a reverse genetics system for Japanese encephalitis virus. AB - Japanese encephalitis virus (JEV) is a common agent of viral encephalitis that causes high mortality and morbidity among children. Molecular genetic studies of JEV are hampered by the lack of a genetically stable full-length infectious JEV cDNA clone. We describe here the development of such a clone. A JEV isolate was fully sequenced, and then its full-length cDNA was cloned into a bacterial artificial chromosome. This was then further engineered so that transcription of the cDNA in vitro would generate synthetic RNAs with authentic 5' and 3' ends. The synthetic RNAs thus produced were highly infectious in susceptible cells (>10(6) PFU/ micro g), and these cells rapidly generated a high titer of synthetic viruses (>5 x 10(6) PFU/ml). The recovered viruses were indistinguishable from the parental virus in terms of plaque morphology, growth kinetics, RNA accumulation, protein expression, and cytopathogenicity. Significantly, the structural and functional integrity of the cDNA was maintained even after 180 generations of growth in Escherichia coli. A single point mutation acting as a genetic marker was introduced into the cDNA and was found in the genome of the recovered virus, indicating that the cDNA can be manipulated. Furthermore, we showed that JEV is an attractive vector for the expression of heterologous genes in a wide variety of cell types. This novel reverse genetics system for JEV will greatly facilitate research into JEV biology. It will also be useful as a heterologous gene expression vector and will aid the development of a vaccine against JEV. PMID- 12743303 TI - Characterization of a conformational epitope on hepatitis B virus core antigen and quasiequivalent variations in antibody binding. AB - We have characterized a conformational epitope on capsids of hepatitis B virus (HBV) by cryo-electron microscopy and three-dimensional image reconstruction of Fab-labeled capsids to approximately 10-A resolution, combined with molecular modeling. The epitope straddles the interface between two adjacent subunits and is discontinuous, consisting of five peptides-two on one subunit and three on its neighbor. Together, the two icosahedral forms of the HBV capsid-T=3 and T=4 particles-present seven quasiequivalent variants of the epitope. Of these, only three bind this Fab. Occupancy ranges from approximately 100 to approximately 0%, reflecting conformational variations in the epitope and steric blocking effects. In the former, small shifts of the component peptides have large effects on binding affinity. This approach appears to hold general promise for elucidating conformational epitopes of HBV and other viruses, including those of neutralizing and diagnostic significance. PMID- 12743304 TI - Host range of Kaposi's sarcoma-associated herpesvirus in cultured cells. AB - Difficulties in efficiently propagating Kaposi's sarcoma-associated herpesvirus (KSHV) in culture have generated the impression that the virus displays a narrow host range. Here we show that, contrary to expectation, KSHV can establish latent infection in many adherent cell lines, including human and nonhuman cells of epithelial, endothelial, and mesenchymal origin. (Paradoxically, the only lines in which we have not observed successful latent infection are cultured lymphoma cell lines.) In most latently infected lines, spontaneous lytic replication is rare and (with only two exceptions) is not efficiently induced by phorbol ester treatment-a result that explains the failure of most earlier studies to observe efficient serial transfer of infection. However, ectopic expression of the KSHV lytic switch protein RTA from an adenoviral vector leads to the prompt induction of lytic replication in all latently infected lines, with the production of infectious KSHV virions. These results indicate (i) that the host cell receptor(s) and entry machinery for KSHV are widely distributed on cultured adherent cells, (ii) that latency is the default pathway of infection, and (iii) that blocks to lytic induction are frequent and largely reside at or upstream of the expression of KSHV RTA. PMID- 12743306 TI - Cellular gene expression survey of vaccinia virus infection of human HeLa cells. AB - Vaccinia virus (VV) is a cytocidal virus that causes major changes in host cell machinery shortly after infecting cells. To define the consequences of virus infection on host gene expression, we used microarrays of approximately 15,000 human cDNAs to examine expression levels of mRNAs isolated at 2, 6, and 16 h postinfection from cultures of infected HeLa cells. The majority of profiling changes during VV infection corresponded to downregulation of genes at 16 h postinfection. Differentially expressed genes were clustered into seven groups to identify common regulatory pathways, with most of them (90%) belonging to clusters 6 and 7, which represent genes whose expression was repressed after infection. Cluster 1, however, contained 37 transcripts (2.81%) showing a robust pattern of induction that was maintained during the course of infection. Genes in cluster 1 included those for Wiskott-Aldrich syndrome protein (WASP) family member WASF1, thymosine, adenosine A2a receptor, glutamate decarboxylase 2, CD-80 antigen, KIAA0888 protein, selenophosphate synthetase, pericentrin, and attractin as well as several expressed sequence tags. We analyzed in more detail the fate of WASP protein in VV-infected cells, because a related family member, N-WASP, is involved in viral motility. WASP protein accumulated in the course of infection; its increase required viral DNA replication and de novo protein synthesis, and it localized in cytoplasmic structures distinct from uninfected cells. This study is the first quantitative analysis of host gene expression following VV infection of cultured human cells, demonstrating global changes in the expression profile, and identifies upregulated genes with potential roles in the virus replication cycle. PMID- 12743305 TI - Molecular and genetic determinants of rous sarcoma virus integrase for concerted DNA integration. AB - Site-directed mutagenesis of recombinant Rous sarcoma virus (RSV) integrase (IN) allowed us to gain insights into the protein-protein and protein-DNA interactions involved in reconstituted IN-viral DNA complexes capable of efficient concerted DNA integration (termed full-site). At 4 nM IN, wild-type (wt) RSV IN incorporates approximately 30% of the input donor into full-site integration products after 10 min of incubation at 37 degrees C, which is equivalent to isolated retrovirus preintegration complexes for full-site integration activity. DNase I protection analysis demonstrated that wt IN was able to protect the viral DNA ends, mapping approximately 20 bp from the end. We had previously mapped the replication capabilities of several RSV IN mutants (A48P and P115S) which appeared to affect viral DNA integration in vivo. Surprisingly, recombinant RSV A48P IN retained wt IN properties even though the virus carrying this mutation had significantly reduced integrated viral DNA in comparison to wt viral DNA in virus-infected cells. Recombinant RSV P115S IN also displayed all of the properties of wt RSV IN. Upon heating of dimeric P115S IN in solution at 57 degrees C, it became apparent that the mutation in the catalytic core of RSV IN exhibited the same thermolabile properties for 3' OH processing and strand transfer (half-site and full-site integration) activities consistent with the observed temperature-sensitive defect for integration in vivo. The average half life for inactivation of the three activities were similar, ranging from 1.6 to 1.9 min independent of the IN concentrations in the assay mixtures. Wt IN was stable under the same heat treatment. DNase I protection analysis of several conservative and nonconservative substitutions at W233 (a highly conserved residue of the retrovirus C-terminal domain) suggests that this region is involved in protein-DNA interactions at the viral DNA attachment site. Our data suggest that the use of recombinant RSV IN to investigate efficient full-site integration in vitro with reference to integration in vivo is promising. PMID- 12743307 TI - Defects in human immunodeficiency virus budding and endosomal sorting induced by TSG101 overexpression. AB - Retrovirus budding is greatly stimulated by the presence of Gag sequences known as late or L domains. The L domain of human immunodeficiency virus type 1 (HIV-1) maps to a highly conserved Pro-Thr-Ala-Pro (PTAP) sequence in the p6 domain of Gag. We and others recently observed that the p6 PTAP motif interacts with the cellular endosomal sorting protein TSG101. Consistent with a role for TSG101 in virus release, we demonstrated that overexpressing the N-terminal, Gag-binding domain of TSG101 (TSG-5') suppresses HIV-1 budding by blocking L domain function. To elucidate the role of TSG101 in HIV-1 budding, we evaluated the significance of the binding between Gag and TSG-5' on the inhibition of HIV-1 release. We observed that a mutation in TSG-5' that disrupts the Gag/TSG101 interaction suppresses the ability of TSG-5' to inhibit HIV-1 release. We also determined the effect of overexpressing a panel of truncated TSG101 derivatives and full-length TSG101 (TSG-F) on virus budding. Overexpressing TSG-F inhibits HIV-1 budding; however, the effect of TSG-F on virus release does not require Gag binding. Furthermore, overexpression of the C-terminal portion of TSG101 (TSG-3') potently inhibits budding of not only HIV-1 but also murine leukemia virus. Confocal microscopy data indicate that TSG-F and TSG-3' overexpression induces an aberrant endosome phenotype; this defect is dependent upon the C-terminal, Vps-28-binding domain of TSG101. We propose that TSG-5' suppresses HIV-1 release by binding PTAP and blocking HIV-1 L domain function, whereas overexpressing TSG-F or TSG-3' globally inhibits virus release by disrupting the cellular endosomal sorting machinery. These results highlight the importance of TSG101 and the endosomal sorting pathway in virus budding and suggest that inhibitors can be developed that, like TSG-5', target HIV-1 without disrupting endosomal sorting. PMID- 12743308 TI - Activation of fusion by the SER virus F protein: a low-pH-dependent paramyxovirus entry process. AB - SER virus, a paramyxovirus closely related to simian virus 5, induces no syncytium formation. The SER virus F protein has a long cytoplasmic tail (CT), and truncation or mutations of the CT result in enhanced syncytium formation (S. Seth, A. Vincent, and R. W. Compans, J. Virol. 77:167-178, 2003; S. Tong, M. Li, A. Vincent, R. W. Compans, E. Fritsch, R. Beier, C. Klenk, M. Ohuchi, and H.-D. Klenk, Virology 301:322-333, 2002). We hypothesized that the presence of the long CT serves to stabilize the metastable conformation of the F protein. We observed that the hemifusion, cytoplasmic content mixing, and syncytium formation ability of the wild-type SER virus F coexpressed with the SER virus hemagglutinin neuraminidase (HN) protein was enhanced, both qualitatively and quantitatively, at elevated temperatures. We also observed enhanced hemifusion, content mixing, and syncytium formation in SER virus F- and HN-expressing cells at reduced pH conditions ranging between 4.8 and 6.2. We have obtained evidence that in contrast to other paramyxoviruses, entry of SER virus into cells occurs by a low pH-dependent process, indicating that the conversion to the fusion-active state for SER virus F is triggered by exposure to reduced pH. PMID- 12743309 TI - Eggplant latent viroid, the candidate type species for a new genus within the family Avsunviroidae (hammerhead viroids). AB - Viroids, small circular RNAs that replicate independently and in most cases incite diseases in plants, are classified into the families Pospiviroidae, composed of species with a central conserved region (CCR) and without hammerhead ribozymes, and Avsunviroidae, composed of three members lacking CCR but able to self-cleave in both polarity strands through hammerhead ribozymes. Here we report the biological and molecular properties of Eggplant latent viroid (ELVd). Purified circular ELVd induces symptomless infections when inoculated into eggplant seedlings. ELVd can be transmitted horizontally and through seed. Sequencing 10 complete cDNA clones showed that ELVd is a circular RNA of 332 to 335 nucleotides with high variability. This RNA can adopt a quasi-rod-like secondary structure of minimal free energy and alternative foldings that permit formation of stable hammerhead structures in plus and minus strands. The ribozymes are active in vitro and, most likely, in vivo. Considering the ELVd properties to be intermediate between those of the two genera of family Avsunviroidae, we propose ELVd as the type species of a third genus with the name ELAVIROID: PMID- 12743310 TI - Activation of adenovirus early promoters and lytic phase in differentiated strata of organotypic cultures of human keratinocytes. AB - Human oncolytic adenoviruses have been used in clinical trials targeting cancers of epithelial origin. To gain a better understanding of the infectious cycle of adenovirus in normal human squamous tissues, we examined the viral infection process in organotypic cultures of primary human keratinocytes. We show that for the infection to occur, wounding of the epithelium is required. In addition, infection appears to initiate at the basal or parabasal cells that express the high-affinity coxsackievirus-adenovirus receptor, CAR, whereas the productive phase takes place in differentiated cells. This is due, at least in part, to the differentiation-dependent activation of the E1A and E2A early promoters and E4 promoters. We also show that adenovirus infection triggers a response mediated by the abnormal accumulation of cyclin E and p21cip1 proteins similar to the one previously observed in human papillomavirus-infected tissues. However, the virus seems to be able to overcome it, at least partially. PMID- 12743311 TI - Excretion of wild-type and vaccine-derived poliovirus in the feces of poliovirus receptor-transgenic mice. AB - The emergence of circulating vaccine-derived poliovirus (cVDPV) strains in suboptimally vaccinated populations is a serious threat to the global poliovirus eradication. The genetic determinants for the transmissibility phenotype of polioviruses, and in particularly of cVDPV strains, are currently unknown. Here we describe the fecal excretion of wild-type poliovirus, oral polio vaccine, and cVDPV (Hispaniola) strains after intraperitoneal injection in poliovirus receptor transgenic mice. Both the pattern and the level of fecal excretion of the cVDPV strains resemble those of wild-type poliovirus type 1. In contrast, very little poliovirus was present in the feces after oral polio vaccine administration. This mouse model will be helpful in elucidating the genetic determinants for the high fecal-oral transmission phenotype of cVDPV strains. PMID- 12743312 TI - Multiple Epstein-Barr virus infections in healthy individuals. AB - We employed a newly developed genotyping technique with direct representational detection of LMP-1 gene sequences to study the molecular epidemiology of Epstein Barr virus (EBV) infection in healthy individuals. Infections with up to five different EBV genotypes were found in two of nine individuals studied. These results support the hypothesis that multiple EBV infections of healthy individuals are common. The implications for the development of an EBV vaccine are discussed. PMID- 12743313 TI - Cell-free synthesis of encephalomyocarditis virus. AB - We developed a system for complete replication of encephalomyocarditis virus (EMCV) in a test tube by using an in vitro translation extract from Krebs-2 cells. Efficient virus synthesis occurred in a narrow range of Mg(2+) and EMCV RNA concentrations. Excess input RNA impaired RNA replication and virus production but not translation. This suggests the existence of a negative feedback mechanism for regulation of RNA replication by the viral plus-strand RNA or proteins. PMID- 12743314 TI - Identification of herpes simplex virus type 1 latency-associated transcript sequences that both inhibit apoptosis and enhance the spontaneous reactivation phenotype. AB - The herpes simplex virus type 1 (HSV-1) latency-associated transcript (LAT) gene is essential for the high spontaneous and induced reactivation phenotype of HSV-1 in the rabbit ocular model and for the high induced reactivation phenotype in the mouse ocular model. Recently we showed that LAT has an antiapoptosis function, and we hypothesized that LAT's ability to inhibit apoptosis played an important role in LAT's ability to enhance the reactivation phenotype. Expression of just the first 1.5 kb of the 8.3-kb LAT gene is sufficient for both inhibition of apoptosis in an in vitro transient-transfection assay and the high spontaneous reactivation phenotype in vivo. Here we show the results of more complex mapping studies in which inhibition of apoptosis and the enhanced spontaneous reactivation phenotype also appear to be linked. The HSV-1 mutant virus dLAT371 has a high spontaneous reactivation phenotype in rabbits, suggesting that the LAT region deleted in this mutant (LAT nucleotides 76 to 447) is not required for this phenotype. The LAT3.3A viral mutant (which expresses LAT nucleotides 1 to 1499) also has a high spontaneous reactivation phenotype, suggesting that the region of LAT not expressed by this mutant (LAT nucleotide 1500 to the end of LAT) is also not required for this phenotype. Surprisingly, LAT2.9A, which is a combination of dLAT371 and LAT3.3A (i.e., it expresses LAT nucleotides 1 to 76 and 447 to 1499), has a low spontaneous reactivation phenotype indistinguishable from that of LAT null mutants. We report here that consistent with the low spontaneous reactivation phenotype of LAT2.9A, a plasmid expressing the identical LAT RNA did not inhibit caspase 9-induced apoptosis. In contrast, plasmids containing the same deletion but able to transcribe up to or past LAT nucleotide 2850 (rather than just up to LAT nucleotide 1499) inhibited caspase 9-induced apoptosis, consistent with the high spontaneous reactivation phenotype of dLAT371. Thus, LAT2.9A may have a low spontaneous reactivation phenotype because the LAT RNA that is made cannot block apoptosis, and dLAT371 apparently has a high spontaneous reactivation phenotype because the LAT RNA made has significant antiapoptosis activity. Furthermore, LAT appeared to have at least two regions capable of interfering with caspase 9-induced apoptosis. One region partially overlaps LAT nucleotides 76 to 447. The second region is partially (or completely) downstream of LAT nucleotide 1499. PMID- 12743315 TI - Extensive cross-reactivity of CD4+ adenovirus-specific T cells: implications for immunotherapy and gene therapy. AB - Adenovirus (Ad)-specific T-cell responses in healthy adult donors were investigated. Ad5, inactivated by methylene blue plus visible light, induced proliferation and gamma interferon (IFN-gamma) production in peripheral blood mononuclear cells of the majority of donors. Responding T cells were CD4(+) and produced IFN-gamma upon restimulation with infectious Ad5 and Ads of different subgroups. T-cell clones showed distinct cross-reactivity patterns recognizing Ad serotypes from either one subgroup (C), two subgroups (B and C), or three subgroups (A, B, and C). This cross-reactivity of Ad-specific T cells has relevance both for Ad-based gene therapy protocols, as well as for the feasibility of T-cell-mediated adoptive immunotherapy in recipients of an allogeneic stem cell transplantation. PMID- 12743316 TI - The herpes simplex virus 1 US3 protein kinase blocks caspase-dependent double cleavage and activation of the proapoptotic protein BAD. AB - An earlier report showed that the U(S)3 protein kinase blocked the apoptosis induced by the herpes simplex virus 1 (HSV-1) d120 mutant at a premitochondrial stage. Further studies revealed that the kinase also blocks programmed cell death induced by the proapoptotic protein BAD. Here we report the effects of the U(S)3 protein kinase on the function and state of a murine BAD protein. Specifically, (i) in uninfected cells, BAD was processed by at least two proteolytic cleavages that were blocked by a general caspase inhibitor. The untreated transduced cells expressed elevated caspase 3 activity. (ii) In cells cotransduced with the U(S)3 protein kinase, the BAD protein was not cleaved and the caspase 3 activity was not elevated. (iii) Inasmuch as the U(S)3 protein kinase blocked the proapoptotic activity and cleavage of a mutant (BAD3S/A) in which the codons for the regulatory serines at positions 112, 136, and 155 were each replaced with alanine codons, the U(S)3 protein kinase does not act by phosphorylation of these sites nor was the phosphorylation of these sites required for the antiapoptotic function of the U(S)3 protein kinase. (iv) The U(S)3 protein kinase did not enable the binding of the BAD3S/A mutant to the antiapoptotic proteins 14-3-3. Finally, (v) whereas cleavage of BAD at ASP56 and ASP61 has been reported and results in the generation of a more effective proapoptotic protein with an M(r) of 15,000, in this report we also show the existence of a second caspase dependent cleavage site most likely at the ASP156 that is predicted to inactivate the proapoptotic activity of BAD. We conclude that the primary effect of U(S)3 was to block the caspases that cleave BAD at either residue 56 or 61 predicted to render the protein more proapoptotic or at residue 156, which would inactivate the protein. PMID- 12743317 TI - Viral internal ribosome entry site structures segregate into two distinct morphologies. AB - An increasing number of viruses have been shown to initiate protein synthesis by a cap-independent mechanism involving internal ribosome entry sites (IRESs). Predictions of the folding patterns of these RNA motifs have been based primarily on sequence and biochemical analyses. Biophysical confirmation of the models has been achieved only for the IRES of hepatitis C virus (HCV), which adopts an open structure consisting of two major stems. We have conducted an extensive comparison of flavivirus and picornavirus IRES elements by negative stain transmission electron microscopy. All of the flavivirus IRESs we examined (those of GB virus-B, GB virus-C, and classical swine fever virus) fold to give a structure similar to that of the HCV IRES, as does an IRES recently found on mRNA encoded by human herpesvirus 8. The larger picornavirus IRESs (those of foot-and mouth disease virus, rhinovirus, encephalomyocarditis virus, and hepatitis A virus) are morphologically similar, comprising a backbone with two protruding stems, and distinct from the flavivirus IRESs. PMID- 12743318 TI - The G glycoprotein of respiratory syncytial virus depresses respiratory rates through the CX3C motif and substance P. AB - Respiratory syncytial virus (RSV) infection in the neonate can alter respiratory rates, i.e., lead to episodes of apnea. We show that RSV G glycoprotein reduces respiratory rates associated with the induction of substance P (SP) and G glycoprotein-CX3CR1 interaction, an effect that is inhibited by treatment with anti-G glycoprotein, anti-SP, or anti-CX3CR1 monoclonal antibodies. These data suggest new approaches for treating some aspects of RSV disease. PMID- 12743319 TI - Manipulation of human pluripotent embryonal carcinoma stem cells and the development of neural subtypes. AB - There are few reliable cell systems available to study the process of human neural development. Embryonal carcinoma (EC) cells are pluripotent stem cells derived from teratocarcinomas and offer a robust culture system to research cell differentiation in a manner pertinent to embryogenesis. Here, we describe the recent development of a series of culture procedures that together can be used to induce the differentiation of human EC stem cells, resulting in the formation of either pure populations of differentiated neurons, populations of differentiated astrocytes, or populations of immature neuronal cell types. Cell-type-specific markers were used to examine the induction of EC stem cell differentiation by retinoic acid. In direct response to manipulation of the culture environment, the expression of cell type markers correlated with the differentiation and appearance of distinct neural cell types, including neurons and astrocytes. These experiments demonstrate that cultured human EC stem cells provide a robust model cell system capable of reproducibly forming neural subtypes for research purposes. PMID- 12743320 TI - Selective ablation of human embryonic stem cells expressing a "suicide" gene. AB - Over the past few years, technological procedures have been developed for utilizing stem cells in transplantation medicine. Human embryonic stem (ES) cells can produce an unlimited number of differentiated cells and are, therefore, considered a potential source of cellular material for use in transplantation medicine. However, serious clinical problems can arise when uncontrolled cell proliferation occurs following transplantation. To avoid these potential problems, we genetically engineered human ES cell lines to express the herpes simplex virus thymidine kinase (HSV-tk) gene. Expression of the HSV-tk protein renders the ES cells sensitive to the U.S. Food and Drug Administration-approved drug ganciclovir, inducing destruction of HSV-tk(+) cells at ganciclovir concentrations that are nonlethal to other cell types. The reversion rate of engineered cells was low even under prolonged selection with ganciclovir. The HSV tk(+) clones retained a normal karyotype and the ability to differentiate to cells from all three germ layers. Most importantly, tumors that arose in mice following subcutaneous injection of HSV-tk(+) human ES cells could be ablated in vivo by administration of ganciclovir. By utilizing these cell lines, safety levels can be improved in transplantations involving tissues derived from human ES cells. PMID- 12743321 TI - Delayed addition of deoxycytidine protects normal CD34+ cells against cytotoxicity of gemcitabine without compromising its activity against human leukemic cells. AB - In phase I and II clinical trials, the deoxycytidine analogue 2',2' difluorodeoxycytidine (dFdC, gemcitabine) has shown promising antitumor activity in leukemia as well as in solid tumors. Preclinical and clinical studies of gemcitabine suggested that myelosuppression was the dose-limiting toxicity. The present investigations were designed to test the effect of continuously administered gemcitabine on the in vitro clonal growth of normal CD34(+) cells isolated from peripheral blood and the promyelocytic cell line, HL-60. For this purpose, CD34(+) and HL-60 cells were cultured in methylcellulose in the continuous presence of 0.1-16 nM of gemcitabine. The results show a dose dependent inhibition of colony growth of normal as well as leukemic cells. However, HL-60 cells were up to 12-fold more sensitive towards gemcitabine than normal progenitors. For rescue experiments, the natural pyrimidine deoxycytidine (dCyd) was added to CD34(+) and HL-60 cells simultaneously or with delay. Coadministration of 1mM dCyd to separate cultures resulted in complete restoration of colony formation capacity of CD34(+) and HL-60 cells. Delayed addition of 1 mM dCyd after 48 and 72 hours recovered up to 90% and 40%, respectively, of stem cell proliferation, whereas HL-60 cells remained substantially inhibited (4.5% +/- 3.5% versus 0%). Delayed addition after 48 and 72 hours protected about 80% and 50%, respectively, of myelopoietic and erythropoetic colony formation, whereas colony formation obtained from HL-60 cells remained significantly inhibited (9.6% +/- 4.17% versus 0%). These in vitro data suggest that there is a marked difference in the susceptibility of leukemic and normal CD34(+) cells to gemcitabine and that delayed administration of dCyd may further reduce the bone marrow cytotoxicity of gemcitabine without impairing its antitumor effect. PMID- 12743322 TI - A role for thrombopoietin in hemangioblast development. AB - Vascular endothelial growth factor (VEGF) and stem cell factor (SCF) act as growth factors for the hemangioblast, an embryonic progenitor of the hematopoietic and endothelial lineages. Because thrombopoietin (TPO) and its receptor, c-Mpl, regulate primitive hematopoietic populations, including bone marrow hematopoietic stem cells, we investigated whether TPO acts on the hemangioblasts that derive from differentiation of embryonic stem cells in vitro. Reverse transcriptase polymerase chain reaction analysis detected expression of c Mpl beginning on day 3 of embryoid body differentiation when the hemangioblast first arises. In assays of the hemangioblast colony-forming cell (BL-CFC), TPO alone supported BL-CFC formation and nearly doubled the number of BL-CFC when added together with VEGF and SCF. When replated under the appropriate conditions, TPO-stimulated BL-CFC gave rise to secondary hematopoietic colonies, as well as endothelial cells, confirming their nature as hemangioblasts. Addition of a neutralizing anti-VEGF antibody did not block TPO enhancement of BL-CFC formation, suggesting that TPO acts independently of VEGF. These results establish that Mpl signaling plays a role in the earliest stages of hematopoietic development and that TPO represents a third growth factor influencing hemangioblast formation. PMID- 12743323 TI - Multilineage differentiation of rhesus monkey embryonic stem cells in three dimensional culture systems. AB - In the course of normal embryogenesis, embryonic stem (ES) cells differentiate along different lineages in the context of complex three-dimensional (3D) tissue structures. In order to study this phenomenon in vitro under controlled conditions, 3D culture systems are necessary. Here, we studied in vitro differentiation of rhesus monkey ES cells in 3D collagen matrixes (collagen gels and porous collagen sponges). Differentiation of ES cells in these 3D systems was different from that in monolayers. ES cells differentiated in collagen matrixes into neural, epithelial, and endothelial lineages. The abilities of ES cells to form various structures in two chemically similar but topologically different matrixes were different. In particular, in collagen gels ES cells formed gland like circular structures, whereas in collagen sponges ES cells were scattered through the matrix or formed aggregates. Soluble factors produced by feeder cells or added to the culture medium facilitated ES cell differentiation into particular lineages. Coculture with fibroblasts in collagen gel facilitated ES cell differentiation into cells of a neural lineage expressing nestin, neural cell adhesion molecule, and class III beta-tubulin. In collagen sponges, keratinocytes facilitated ES cell differentiation into cells of an endothelial lineage expressing factor VIII. Exogenous granulocyte-macrophage colony stimulating factor further enhanced endothelial differentiation. Thus, both soluble factors and the type of extracellular matrix seem to be critical in directing differentiation of ES cells and the formation of tissue-like structures. Three-dimensional culture systems are a valuable tool for studying the mechanisms of these phenomena. PMID- 12743325 TI - In utero hematopoietic stem cell transplantation in nonhuman primates: the role of T cells. AB - In utero transplantation of hematopoietic stem cells is a promising treatment for immune and hematologic diseases of fetuses and newborns. Unfortunately, there are limited data from nonhuman primates and humans describing optimal transplantation conditions. The purpose of this investigation was to determine the effect of T cell number on engraftment and the level of chimerism after in utero transplantation in nonhuman primates. CD34(+) allogeneic adult bone marrow cells, obtained from the sire after G-CSF and stem cell factor administration, were transplanted into female fetal recipients. The average CD34(+) cell dose was 3.0 x 10(9)/kg (range, 9.9 x 10(8) to 4.4 x 10(9)) and the T-cell dose ranged from 2.6 x 10(5) to 1.1 x 10(8)/kg. Chimerism was determined in peripheral blood subsets (CD2, CD13, and CD20) and in progenitor cell populations by using polymerase chain reaction. Chimerism was noted in seven of eight live-born animals. The level of chimerism in the progenitor population was related to the fetal T-cell dose (r = 0.64, p < 0.02). At the lowest T-cell dose (2.6 x 10(5)/kg), no chimerism was detected. As the T-cell dose increased to 10(6-7)/kg, the level of chimerism increased. Adjusting the T-cell dose to 1.1 x 10(8)/kg resulted in fatal graft-versus-host disease (GVHD). The results of this study emphasize the importance of T cells in facilitating donor cell engraftment and in producing GVHD in fetal nonhuman primates. Some animals achieved levels of chimerism in the marrow hematopoietic progenitor cell population that would likely have clinical relevance. However, the levels of chimerism in peripheral blood were too low for therapeutic benefit. Further studies are needed to test methods that are likely to enhance donor cell engraftment and peripheral blood levels of donor cells. PMID- 12743324 TI - Absolute values of dendritic cell subsets in bone marrow, cord blood, and peripheral blood enumerated by a novel method. AB - Dendritic cells (DCs) are pivotal in inducing immunity or alternatively downregulating immune reactivity. In humans, the opposing phenotypic subsets of CD11c(+)/CD123(-) "myeloid" DCs and CD123(+)/CD11c(-) "lymphoid" DCs have been proposed to orchestrate these immune responses. In this study we determined the absolute numbers of both subsets in three resting hematopoietic tissues by employing a novel flow cytometry method, eliminating processing steps and calculations based on mononuclear cell percentages. Internal bead standards along with the cells of interest were simultaneously acquired directly from unmanipulated whole blood specimens. We found significant differences (p < 0.001) between the mean absolute numbers of CD123(+)/CD11c(-) lymphoid DCs among the three sources, with the fewest present in peripheral blood (8.2/ micro l), about 50% more in cord blood (12.2/ micro l), and fivefold more in bone marrow (40.2/ micro l). Cord blood and bone marrow CD11c(+)/CD123(-) myeloid DC counts did not differ from each other (23.5/ micro l and 28.9/ micro l, respectively) but peripheral blood contained significantly fewer (15.5/ micro l, p = 0.006). CD11c(+)/CD123(-) DCs had significantly higher surface expression of HLA-DR (p < 0.001) in all three sources. To test for association with the DC subsets, T, B, and natural killer (NK) lymphocytes were also enumerated. In bone marrow only, significant correlations were found between the size of the CD123(+)/CD11c(-) lymphoid DC pool and NK cells (p = 0.0029) and B cells (p = 0.0033). These data support the hypothesis that a common CD123(+)/CD11c(-) DC, NK cell, and B cell progenitor is resident in marrow, and this cell may be identical to the common lymphoid progenitor previously described in mice and/or the human CD34(+)/Lin( )/CD10(+) progenitor. PMID- 12743326 TI - Analysis of gene expression profiles in an imatinib-resistant cell line, KCL22/SR. AB - The BCR/ABL tyrosine kinase inhibitor, imatinib, has shown substantial effects in blast crises of chronic myelogenous leukemia. However, most patients relapse after an initial clinical response, indicating that drug resistance is a major problem for patients being treated with imatinib. In this study, we generated a new imatinib-resistant BCR/ABL-positive cell line, KCL22/SR. The 50% inhibitory concentration of imatinib was 11-fold higher in KCL22/SR than in the imatinib sensitive parental cell line, KCL22. However, KCL22/SR showed no mutations in the BCR/ABL gene and no increase in the levels of BCR/ABL protein and P-glycoprotein. Furthermore, the level of phosphorylated BCR/ABL protein was suppressed by imatinib treatment, suggesting that mechanisms independent of BCR/ABL signaling are involved in the imatinib resistance in KCL22/SR cells. DNA microarray analyses demonstrated that the signal transduction-related molecules, RAS p21 protein activator and RhoA, which could affect Ras signaling, and a surface tumor antigen, L6, were upregulated, while c-Myb and activin A receptor were downregulated in KCL22/SR cells. Furthermore, imatinib treatment significantly suppressed the level of phosphorylated p44/42 in KCL22 cells but not in KCL22/SR cells, even when BCR/ABL was inhibited by imatinib. These results suggest that various mechanisms, including disturbance of Ras-mitogen-activated protein kinase signaling, are involved in imatinib resistance. PMID- 12743327 TI - Defining epithelial cell progenitors in the human oxyntic mucosa. AB - In the human stomach, the oxyntic epithelium includes numerous tubular invaginations consisting of short pits opening into long glands. The pit is lined by pit cells, whereas the gland is composed of three regions: the base, containing zymogenic cells; the neck, containing neck cells; and the isthmus, composed of little known immature cells and of parietal cells, which are also scattered through the neck and base. The aim of this study was to examine the ultrastructure of the immature cells and to determine their relation to mature cells. To do so, normal oxyntic mucosal biopsies from subjects ranging from 20-43 years old were fixed in aldehydes and postfixed in reduced osmium for electron microscopy and morphometric analysis. The immature cells were sorted out into four classes, whose roles were clarified by comparison with the thoroughly investigated mouse oxyntic epithelium. The first class was composed of the least differentiated immature cells, which were rare and characterized by minute, dense, or cored secretory granules and were accordingly named mini-granule cells. Their function was not clarified. The second class consisted of pre-pit cells, which were characterized by few dense mucous granules and give rise to pit cells that ascend the pit wall and, after reaching the luminal surface, die or are extruded. Both pre-pit and pit cells underwent continuous renewal and, therefore, together constituted a renewal system referred to as pit cell lineage. The third class, or pre-neck cells, characterized by cored secretory granules, give rise to neck cells that descend toward the base region and differentiate further into pre zymogenic cells, which finally become zymogenic cells. The latter eventually degenerate and die. Thus pre-neck cells and their progeny constitute a renewing system, designated zymogenic cell lineage. The fourth class, or pre-parietal cells, characterized by long microvilli and few tubulovesicles, differentiate into parietal cells that descend along the neck and base regions and eventually degenerate and die. Pre-parietal and parietal cells represent a renewing system referred to as parietal cell lineage. While the origin of the last three classes of progenitor cells has not been elucidated, it is likely that they arise either from an unidentified multipotential stem cell, possibly the mini-granule cell itself, or from the mitotic activity of pre-pit and pre-neck cells. In conclusion, the human oxyntic epithelium is composed of continually renewing cells organized in distinct cell lineages. PMID- 12743328 TI - Hypoxia promotes murine bone-marrow-derived stromal cell migration and tube formation. AB - Recent evidence indicates that bone-marrow-derived stromal cells (MSCs) have a histology coherent with endothelial cells that may enable them to contribute to tumor angiogenesis through yet undefined mechanisms. In this work, we investigated the angiogenic properties of murine MSCs involved in extracellular matrix degradation and in neovascularization that could take place in a hypoxic environment such as that encountered in tumor masses. MSCs were cultured in normoxia (95% air and 5% CO(2)) or in hypoxia (1% oxygen, 5% CO(2), and 94% nitrogen). We found that hypoxic culture conditions rapidly induced MSC migration and three-dimensional capillary-like structure formation on Matrigel. In vitro, MSC migration was induced by growth-factor- and cytokine-enriched conditioned media isolated from U-87 glioma cells as well as from MSCs cultured in hypoxic conditions, suggesting both paracrine and autocrine regulatory mechanisms. Although greater vascular endothelial growth factor levels were secreted by MSCs in hypoxic conditions, this growth factor alone could not explain their greater migration. Interestingly, matrix metalloproteinase (MMP)-2 mRNA expression and protein secretion were downregulated, while those of membrane-type (MT)1-MMP were strongly induced by hypoxia. Functional inhibition of MT1-MMP by a blocking antibody strongly suppressed MSC ability to migrate and generate capillary-like structures. Collectively, these data suggest that MSCs may have the capacity to participate in tumor angiogenesis through regulation of their angiogenic properties under an atmosphere of low oxygen that closely approximates the tumor microenvironment. PMID- 12743329 TI - Cell therapy: filling the gap between basic science and clinical trials October 15-17, 2001, Rome, Italy. AB - Summarized here, and in forthcoming issues of, are the concepts that emerged at a recent international workshop on cell therapy organized by The Istituto Superiore di Sanita in Rome in collaboration with Istituto Dermatopatico dell'Immacolata, Rome; Istituto Nazionale Ricerca Cancro-Centro Biotecnologie Avanzate, Genova; and University G. D'Annunzio, Chieti. The meeting intent was to provide an overview of the most recent developments in cell therapy, the future perspectives for these clinical trials, and the regulatory issues they involve, as well as a progress report on the clinical protocols that have been approved up to now in Italy. The meeting included six scientific sessions (Immunotherapy, Epithelium, Osteoregeneration, Hematopoiesis, Future Perspectives, and Overview of the National and International Regulations) and involved lectures from Italian and foreign scientists. PMID- 12743330 TI - A new type I IFN-mediated pathway for the rapid differentiation of monocytes into highly active dendritic cells. AB - Dendritic cells (DCs) are a unique leukocyte type consisting of different subsets of professional antigen-presenting cells. Since DCs initiate and govern the immune response, they represent an ideal target for intervention aimed at modulating and potentiating immune responses against cancer and infectious diseases. We recently described and characterized, at a functional level, a novel DC subset, interferon (IFN)-DCs, derived from blood monocytes after a short exposure to type I IFN and GM-CSF. Here, we review our recent studies on IFN-DCs and discuss their possible use in clinical immunotherapeutic strategies. PMID- 12743332 TI - New approaches to training general practitioners. PMID- 12743331 TI - Expression of functional CXCR4 by muscle satellite cells and secretion of SDF-1 by muscle-derived fibroblasts is associated with the presence of both muscle progenitors in bone marrow and hematopoietic stem/progenitor cells in muscles. AB - We found that the murine cell lines C2C12 and G7 derived from muscle satellite cells, which are essential for muscle regeneration, express the functional CXCR4 receptor on their surface and that the specific ligand for this receptor, alpha chemokine stromal-derived factor 1 (SDF-1), is secreted in muscle tissue. These cell lines responded to SDF-1 stimulation by chemotaxis, phosphorylation of mitogen-activated protein kinase (MAPK) p42/44 and AKT serine-threonine kinase, and calcium flux, confirming the functionality of the CXCR4 receptor. Moreover, supernatants derived from muscle fibroblasts chemoattracted both satellite cells and human CD34(+) hematopoietic stem/progenitor cells. In a similar set of experiments, supernatants from bone marrow fibroblasts were found to chemoattract CXCR4(+) satellite cells just as they chemoattract CD34(+) cells. Moreover, preincubation of both muscle satellite cells and hematopoietic stem/progenitor CD34(+) cells before chemotaxis with T140, a specific CXCR4 inhibitor, resulted in a significantly lower chemotaxis to media conditioned by either muscle- or bone marrow-derived fibroblasts. Based on these observations, we postulate that the SDF-1-CXCR4 axis is involved in chemoattracting circulating CXCR4(+) muscle stem/progenitor and circulating CXCR4(+) hematopoietic CD34(+) cells to both muscle and bone marrow tissues. Thus, it appears that tissue-specific stem cells circulating in peripheral blood could compete for SDF-1(+) niches, and this would explain, without invoking the concept of stem cell plasticity, why hematopoietic colonies can be cultured from muscles and early muscle progenitors can be cultured from bone marrow. PMID- 12743333 TI - Modern treatment of acromegaly. AB - Acromegaly is an endocrine disorder characterised by increased morbidity and mortality. It is usually caused by a growth hormone secreting pituitary adenoma and is manifested by a variety of clinical features. Surgery is usually the treatment of choice, however over the last few years, several new methods of treatment have been developed. A recent consensus on the targets for treatment has led to multiple studies being conducted to assess the efficacy of the currently available options. This review examines the evidence for and against these treatments. PMID- 12743334 TI - Mechanisms of oxidative stress and vascular dysfunction. AB - The endothelium regulates vascular homoeostasis through local elaboration of mediators that modulate vascular tone, platelet adhesion, inflammation, fibrinolysis, and vascular growth. Impaired vascular function contributes to the pathogenesis of atherosclerosis and acute coronary syndromes. There is growing pathophysiological evidence that increased generation of reactive oxygen species and oxidative stress participates in proatherogenic mechanisms of vascular dysfunction and atherothrombosis. In this review, the role of oxidative stress in mechanisms of vascular dysfunction is discussed, and potential antioxidant strategies are reviewed. PMID- 12743335 TI - Impact of respiratory viral infections on cystic fibrosis. AB - The life expectancy for patients with cystic fibrosis has improved remarkably over the last 20 years. Progressive deterioration of pulmonary function continues despite the aggressive use of antimicrobials. The absence of fever, neutrophilia, and systemic symptoms suggest that during pulmonary exacerbations other non bacterial factors may have played a part. Some have suggested respiratory viruses as main suspects. So far, few data have illustrated the relationship of respiratory viruses and cystic fibrosis. By gaining further knowledge of this relationship, one may change future clinical practice and boost the survival of these patients. PMID- 12743336 TI - What should a suspended doctor do when his actions could save a patient's life? AB - Suspension from duty is becoming an increasing threat for medical practitioners. While under suspension a doctor may, in extreme circumstances, find themselves in a difficult legal dilemma as to their duty of care towards others. A suspended doctor may risk litigation if they are unaware of the legal ramifications of their unusual circumstances. The following article outlines the legal situation as it applies to England and Wales. PMID- 12743337 TI - Acute glomerulonephritis. AB - Glomerulonephritis is an important cause of renal failure thought to be caused by autoimmune damage to the kidney. While each type of glomerulonephritis begins with a unique initiating stimulus, subsequent common inflammatory and fibrotic events lead to a final pathway of progressive renal damage. In this article the different forms of inflammatory glomerulonephritis and their diagnosis are discussed. In a review of therapy both immediate life saving treatment given when glomerulonephritis causes acute renal failure and more specific treatments designed to modify the underlying mechanisms of renal injury are considered. PMID- 12743340 TI - Oral vitamin B12 can change our practice. AB - Oral vitamin B12 can provide an effective alternative to intramuscular injections, so giving patients a choice and reducing costs in primary care. This study investigated the effectiveness, safety, and acceptability of oral vitamin B12 as replacement therapy in patients with vitamin B12 deficiency in a city general practice population. Forty patients previously maintained on vitamin B12 injections were given 1000 micro g of oral cyanocobalamin daily for up to 18 months. All the patients maintained satisfactory serum B12 levels and showed normal haematology and neurology. Compliance and acceptability was excellent. The time for a change in practice has indeed arrived. PMID- 12743338 TI - Chest radiographs in the emergency department: is the radiologist really necessary? AB - BACKGROUND: The chest radiograph is considered one of the most complex imaging modalities to interpret. Several studies have evaluated radiograph interpretation in the emergency department, and considerable disagreement among clinical physicians and expert radiologists has been observed in the reading of chest films. The interpretation of chest radiographs by emergency department physicians was compared with senior radiologists in discharged patients, and misinterpretations assessed in relation to the physician's level of training. METHODS: Radiological descriptions of 509 chest radiographs of 507 patients, aged 16-98 years who were discharged from the emergency department, were prospectively reviewed. Missed findings were recorded with regard to the physician's level of training and experience. The effects of misinterpretations on discharge recommendations were also investigated. Statistical assessment was conducted using the chi(2) test. Interobserver agreement was also tested by the kappa coefficient. RESULTS: The sensitivity for detecting different abnormalities in the radiographs ranged from 20% to 64.9% and specificity from 94.9% to 98.7%. Despite the low sensitivities found, there were relatively few clinical implications of the "missed" findings since they were either of a minor nature or appropriate follow up was prescribed. The overall interobserver reliability, assessed by the kappa coefficient, was 0.40 (95% confidence interval 0.35 to 0.46). These findings did not change significantly by emergency department physician's level of training. CONCLUSIONS: Emergency department physicians frequently miss specific radiographic abnormalities and there is considerable discrepancy between their interpretations and those of trained radiologists. These findings highlight the importance of routine evaluation of chest radiographs by a well trained radiologist and emphasise the need for improving interpretive skills among emergency department physicians. PMID- 12743341 TI - Rice bodies in ischiogluteal bursitis. PMID- 12743342 TI - Results of a programme to improve house staff use of metered dose inhalers and spacers. AB - PURPOSE: Metered dose inhalers (MDIs) and spacers are used widely in the treatment of asthma. Medical personnel who are responsible for training patients must themselves be proficient with the devices. The proficiency of a group of new medical interns with use of MDI and spacer devices was determined, and improvement in their use of these devices was sought. SUBJECTS: Fifty six medical interns tested at the start of their first house staff training year. METHODS: The ability of medical interns to use MDIs and spacers was assessed using a visual scoring system before and after a large group lecture emphasising proper device use and once again after an intensive one-on-one training session with an attending physician. RESULTS: Initially, only 5% used an MDI perfectly. This improved to 13% after a lecture and demonstration, and 73% after an intensive one on-one session. Almost no new interns could use a collapsible or tube spacer properly initially. This improved to 15% and 29% respectively after a lecture. After one-on-one training, correct technique was increased to 69% for collapsible spacer and 95% for the tube spacer. Analysis of individual steps of MDI use showed that interns had particular difficulty in coordinating actuation with inhalation. The tube spacer appeared easiest to learn. CONCLUSIONS: Incoming medical house staff have limited ability to use MDI with and without spacers. A large group lecture is relatively ineffective when compared with a one-on-one training session in training with these devices. PMID- 12743344 TI - Iron deficiency anaemia: are the British Society of Gastroenterology guidelines being adhered to? AB - BACKGROUND: The British Society of Gastroenterology (BSG) issued guidelines on the investigation of iron deficiency anaemia (IDA) ensuring standardised and comprehensive gastrointestinal investigation in all patients. It was apparent that not all patients in the authors' hospital were investigated according to these guidelines. OBJECTIVE: To determine whether patients who were referred for upper gastrointestinal endoscopy for investigation of IDA were confirmed to be iron deficient, and whether the BSG guidelines were being fully implemented. METHODS: All patients referred for upper gastrointestinal endoscopy over an 18 month period on a computer database (Endoscribe) were reviewed. Haematology, biochemistry, and radiology results were obtained and the frequency of the various diagnoses recorded. RESULTS: A total of 320 patients (133 male; mean age 71.5 years) were initially referred for upper gastrointestinal endoscopy for investigation of IDA, of whom 95 were iron deficient. Of these, 44 (46%) had duodenal biopsies performed, three (7%) of whom were diagnosed with coeliac disease. Five patients were diagnosed with upper gastrointestinal carcinoma (one oesophageal, four gastric). Of the remaining 87 patients, 65 (75%) underwent lower gastrointestinal investigations with four having colorectal carcinoma, four colonic polyps, and one angiodysplasia. CONCLUSIONS: Duodenal biopsies were performed in less than half of the patients. In those not diagnosed with coeliac disease or upper gastrointestinal carcinoma, only three quarters underwent lower gastrointestinal assessment. Approximately 10% were diagnosed with gastrointestinal malignancy as a cause for their anaemia and in 66% of patients no gastrointestinal cause was found. All physicians need to be made fully aware of the BSG guidelines for investigation of IDA. PMID- 12743345 TI - History of the development of geriatric medicine in the UK. AB - In this review the development of the specialty of geriatric medicine in the UK is traced from its humble beginnings. Elderly medicine is now thriving and represents the largest group of physician members of the Royal Colleges of Physicians. Geriatric medicine is essentially about optimising the care and wellbeing of older people. A key component of this is teamwork. A successful service for old people depends on the skills of many people, including nurses, therapists, social workers, and others. The contributions made by nurses and other professionals have been immense, but space does not permit a historical review of their important role. PMID- 12743346 TI - Iatrogenic injury to the breast bud causing breast hypoplasia. AB - A 14-year-old girl with iatrogenic breast asymmetry is reported. She had multiple operations to the left anterior chest wall in the neonatal period for diaphragmatic hernia that resulted in multiple scars, which damaged the left breast bud. A better thoracotomy incision avoiding the breast bud could prevent later breast hypoplasia. PMID- 12743347 TI - Unusual cause of infective discitis in an adolescent. AB - The first case of infective discitis caused by Eikenella corrodens in an adolescent is presented. The need for anaerobic cultures when dealing with infective pathology in the spine is stressed. A 14 year old boy presented with acute exacerbation of back pain, which showed characteristics of infective discitis after magnetic resonance imaging. Computed tomography guided biopsy grew E corrodens in anaerobic cultures that was sensitive to ampicillin, co-amoxiclav, cefadroxil, and cefotaxime. This patient responded well to co-amoxiclav and recovered without any surgical intervention. PMID- 12743349 TI - Facial and dental appearance of Williams syndrome. PMID- 12743348 TI - Recurrent autumnal psychosis. AB - Acute confusional states in the older patient often have a remediable cause. Every effort should be made to ascertain the cause so that appropriate treatment can be given and future episodes prevented. A patient is described who presented with recurrent episodes of acute psychosis after ingestion of Atropa belladonna (deadly nightshade). PMID- 12743350 TI - A case of reduced consciousness and hypoventilation. PMID- 12743351 TI - Recurrent gastrointestinal bleed. PMID- 12743352 TI - Dyspnoea and a subcutaneous swelling. PMID- 12743353 TI - Acute severe anaemia in an elderly patient with hereditary sphaerocytosis. PMID- 12743359 TI - "Statement on scientific publication and security" fails to provide necessary guidelines. PMID- 12743360 TI - Quality control of ribosomal RNA mediated by polynucleotide phosphorylase and RNase R. AB - Despite their overall accuracy, errors in macromolecular processes, such as rRNA synthesis and ribosome assembly, inevitably occur. However, whether these errors are remediated and how this might be accomplished is not known. In previous work, we showed that a double mutant strain lacking both polynucleotide phosphorylase (PNPase) and RNase R activities is inviable. In the course of examining the molecular basis for this phenotype, we found that shifting a temperature sensitive mutant strain to 42 degrees C led to cessation of growth and loss of cell viability. Northern analysis of RNA isolated from such cells after the temperature shift revealed that fragments of 16S and 23S rRNA accumulated to a high level, and that the amount of ribosomes and ribosomal subunits decreased due to defects in ribosome assembly. rRNA fragments were not detected at 31 degrees C or when single mutant strains were grown at 42 degrees C. Pulse-chase analysis showed that the rRNA fragments appeared within 5 min at 42 degrees C, and that they accumulated before the loss of cell viability. The data are consistent with a model in which PNPase and RNase R mediate a previously unknown quality control process that normally removes defective rRNAs as soon as they are generated. In the absence of these RNases, rRNA fragments accumulate, leading to interference with ribosome maturation and ultimately to cell death. PMID- 12743361 TI - Structural basis of albumin-thyroxine interactions and familial dysalbuminemic hyperthyroxinemia. AB - Human serum albumin (HSA) is the major protein component of blood plasma and serves as a transporter for thyroxine and other hydrophobic compounds such as fatty acids and bilirubin. We report here a structural characterization of HSA thyroxine interactions. Using crystallographic analyses we have identified four binding sites for thyroxine on HSA distributed in subdomains IIA, IIIA, and IIIB. Mutation of residue R218 within subdomain IIA greatly enhances the affinity for thyroxine and causes the elevated serum thyroxine levels associated with familial dysalbuminemic hyperthyroxinemia (FDH). Structural analysis of two FDH mutants of HSA (R218H and R218P) shows that this effect arises because substitution of R218, which contacts the hormone bound in subdomain IIA, produces localized conformational changes to relax steric restrictions on thyroxine binding at this site. We have also found that, although fatty acid binding competes with thyroxine at all four sites, it induces conformational changes that create a fifth hormone-binding site in the cleft between domains I and III, at least 9 A from R218. These structural observations are consistent with binding data showing that HSA retains a high-affinity site for thyroxine in the presence of excess fatty acid that is insensitive to FDH mutations. PMID- 12743363 TI - An actin-ribonucleoprotein interaction is involved in transcription by RNA polymerase II. AB - To determine the function of actin in the cell nucleus, we sought to identify nuclear actin-binding proteins in the dipteran Chironomus tentans using DNase I affinity chromatography. We identified the RNA-binding protein hrp65 as an actin binding protein and showed that the C-terminal sequence of the hrp65-2 isoform is able to interact directly with actin in vitro. In vivo crosslinking and coimmunoprecipitation experiments indicated that hrp65 and actin are also associated in the living cell. Moreover, in vivo administration of a competing peptide corresponding to the C-terminal sequence of hrp65-2 disrupted the actin hrp65-2 interaction and caused a specific and drastic reduction of transcription as judged by puff regression and diminished bromo-UTP incorporation. Our results indicate that an actin-based mechanism is implicated in the transcription of most if not all RNA polymerase II genes and suggest that an actin-hrp65-2 interaction is required to maintain the normal transcriptional activity of the cell. Furthermore, immunoelectron microscopy experiments and nuclear run-on assays suggest that the actin-hrp65-2 complex plays a role in transcription elongation. PMID- 12743362 TI - Calcium-permeable AMPA/kainate receptors mediate toxicity and preconditioning by oxygen-glucose deprivation in oligodendrocyte precursors. AB - Hypoxic-ischemic brain injury in premature infants results in cerebral white matter lesions with prominent oligodendroglial injury and loss, a disorder termed periventricular leukomalacia (PVL). We have previously shown that glutamate receptors mediate hypoxic-ischemic injury to oligodendroglial precursor cells (OPCs) in a model of PVL in the developing rodent brain. We used primary OPC cultures to examine the mechanism of cellular toxicity induced by oxygen-glucose deprivation (OGD) to simulate brain ischemia. OPCs were more sensitive to OGD induced toxicity than mature oligodendrocytes, and OPC toxicity was attenuated by nonselective [2,3-dihydroxy-6-nitro-7-sulfamoylbenzo[f]quinoxaline (NBQX), 6 cyano-7-nitroquinoxaline-2,3-dione], alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA)-preferring (GYKI 52466), kainate-preferring (gamma d-glutamylaminomethanesulfonic acid), or Ca2+-permeable AMPA/kainate receptor antagonists (joro spider toxin, JSTx) administered either during or after OGD. Furthermore, NBQX or JSTx blocked OGD-induced Ca2+ influx. Relevant to recurrent hypoxic-ischemic insults in developing white matter, we examined the effects of sublethal OGD preconditioning. A prior exposure of OPCs to sublethal OGD resulted in enhanced vulnerability to subsequent excitotoxic or OGD-induced injury associated with an increased Ca2+ influx. AMPA/kainate receptor blockade with NBQX or JSTx either during or after sublethal OGD prevented its priming effect. Furthermore, OGD preconditioning resulted in a down-regulation of the AMPA receptor subunit GluR2 on cell surface that increased Ca2+ permeability of the receptors. Overall, these data suggest that aberrantly enhanced activation of Ca2+-permeable AMPA/kainate receptors may be a major mechanism in acute and repeated hypoxic-ischemic injury to OPCs in disorders of developing cerebral white matter, such as PVL. PMID- 12743364 TI - A dual altered peptide ligand down-regulates myasthenogenic T cell responses by up-regulating CD25- and CTLA-4-expressing CD4+ T cells. AB - Immunization of mice with two myasthenogenic peptides, p195-212 and p259-271, which are sequences of the human acetylcholine receptor, resulted in myasthenia gravis (MG)-associated immune responses. A dual altered peptide ligand (APL) composed of the two APLs of the myasthenogenic peptides inhibited, in vitro and in vivo, those responses. The aims of this study were to further elucidate the mechanism/s by which the dual APL down-regulates MG-associated responses in vivo and characterize the cell population/s involved in this immunomodulatory suppressive effect. We have shown here that s.c. administration of the dual APL activates CD4CD25-expressing cells in lymph nodes (LN) of SJL mice. Furthermore, depletion of these cells diminished significantly the inhibitory effect of the APL on p195-212-specific proliferative responses. Depletion of the CD4+CD25+ cells was accompanied with a decrease in the secretion of the immunosuppressive cytokine, transforming growth factor (TGF)-beta. Administration of the dual APL resulted also in the up-regulation of the expression of cytotoxic T lymphocyte antigen (CTLA)-4 and in a down-regulated expression of CD28 on LN cells. Blockade of the CTLA-4 function, in vitro, abrogated the inhibitory effect of the dual APL on the proliferative responses specific to p195-212. Thus, our results suggest that the active suppression exerted by the dual APL is mediated by the CD4+CD25+ immunoregulatory cell population, either directly through the CTLA-4 molecule expressed on these cells, and/or indirectly by causing the differentiation of other regulatory T cell population/s that secrete immunosuppressive cytokines. PMID- 12743365 TI - Direct stimulation of naive T cells by membrane vesicles from antigen-presenting cells: distinct roles for CD54 and B7 molecules. AB - T cell stimulation usually requires direct contact with viable antigen-presenting cells (APCs). However, we show here that small exosome-like membrane vesicles shed from APCs can be recognized by naive CD8+ T cells in the absence of viable APCs. T cell antigen receptor-dependent binding of vesicles by CD8+ cells is MHC class I/peptide-specific and requires that the vesicles coexpress intercellular adhesion molecule 1 (ICAM-1, CD54), although not B7 (B7-1). In the absence of B7, T cell binding of vesicles is nonimmunogenic. By contrast, vesicles expressing both ICAM-1 and B7 are strongly immunogenic and cause purified APC-depleted CD8+ cells to mount peptide-specific proliferative responses and differentiate into effector cells. PMID- 12743366 TI - Golgi-dependent transport of cholesterol to the Chlamydia trachomatis inclusion. AB - Cholesterol, a lipid not normally found in prokaryotes, was identified in purified Chlamydia trachomatis elementary bodies and in the chlamydial parasitophorous vacuole (inclusion) membrane of infected HeLa cells. Chlamydiae obtained eukaryotic host cell cholesterol both from de novo synthesis or low density lipoprotein. Acquisition of either de novo-synthesized cholesterol or low density lipoprotein-derived cholesterol was microtubule-dependent and brefeldin A sensitive, indicating a requirement for the Golgi apparatus. Transport also required chlamydial protein synthesis, indicative of a pathogen-directed process. The cholesterol trafficking pathway appears to coincide with a previously characterized delivery of sphingomyelin to the inclusion in that similar pharmacological treatments inhibited transport of both sphingomyelin and cholesterol. These results support the hypothesis that sphingomyelin and cholesterol may be cotransported via a Golgi-dependent pathway and that the chlamydial inclusion receives cholesterol preferentially from a brefeldin A sensitive pathway of cholesterol trafficking from the Golgi apparatus to the plasma membrane. PMID- 12743367 TI - Molecular genetic analysis of the glycosyltransferase Fringe in Drosophila. AB - Fringe proteins are beta1,3-N-acetylglucosaminyltransferases that modulate signaling through Notch receptors by modifying O-linked fucose on epidermal growth factor domains. Fringe is highly conserved, and comparison among 18 different Fringe proteins from 11 different species identifies a core set of 84 amino acids that are identical among all Fringes. Fringe is only distantly related to other glycosyltransferases, but analysis of the predicted Drosophila proteome identifies a set of four sequence motifs shared among Fringe and other putative beta1,3-glycosyltransferases. To gain functional insight into these conserved sequences, we genetically and molecularly characterized 14 point mutations in Drosophila fringe. Most nonsense mutations act as recessive antimorphs, raising the possibility that Fringe may function as a dimer. Missense mutations identify two distinct motifs that are conserved among beta1,3 glycosyltransferases, and that can be modeled onto key motifs in the crystallographic structures of bovine beta1,4-galactosyltransferase 1 and human glucuronyltransferase I. Other missense mutations map to amino acids that are conserved among Fringe proteins, but not among other glycosyltransferases, and thus may identify structural motifs that are required for unique aspects of Fringe activity. PMID- 12743369 TI - Abnormal phagocytosis by retinal pigmented epithelium that lacks myosin VIIa, the Usher syndrome 1B protein. AB - Mutations in the myosin VIIa gene (MYO7A) cause Usher syndrome type 1B (USH1B), a major type of the deaf-blind disorder, Usher syndrome. We have studied mutant phenotypes in the retinas of Myo7a mutant mice (shaker1), with the aim of elucidating the role(s) of myosin VIIa in the retina and what might underlie photoreceptor degeneration in USH1B patients. A photoreceptor defect has been described. Here, we report that the phagocytosis of photoreceptor outer segment disks by the retinal pigment epithelium (RPE) is abnormal in Myo7a null mice. Both in vivo and in primary cultures of RPE cells, the transport of ingested disks out of the apical region is inhibited in the absence of Myo7a. The results with the cultured RPE cells were the same, irrespective of whether the disks came from wild-type or mutant mice, thus demonstrating that the RPE is the source of this defect. The inhibited transport seems to delay phagosome-lysosomal fusion, as the degradation of ingested disks was slower in mutant RPE. Moreover, fewer packets of disk membranes were ingested in vivo, possibly because retarded removal of phagosomes from the apical processes inhibited the ingestion of additional disk membranes. We conclude that Myo7a is required for the normal processing of ingested disk membranes in the RPE, primarily in the basal transport of phagosomes into the cell body where they then fuse with lysosomes. Because the phagocytosis of photoreceptor disks by the RPE has been shown to be critical for photoreceptor cell viability, this defect likely contributes to the progressive blindness in USH1B. PMID- 12743368 TI - ATP hydrolysis on actin-related protein 2/3 complex causes debranching of dendritic actin arrays. AB - Extension of lamellipodia, an important dissipative process in cell motility, is driven by the turnover of a polarized dendritic array of actin filaments. Motility is driven by catalytic cycles of filament attachment to Wiskott-Aldrich syndrome protein (WASP)-activated actin-related protein (Arp)2/3 complex at the leading edge, branch formation, and detachment, allowing subsequent growth of branched filaments. The morphology, mechanical strength, and lifetime of the array are determined by the processes of filament branching, debranching, and treadmilling. All three processes are controlled by ATP hydrolysis. ATP hydrolysis on F-actin is known to be at the origin of treadmilling. Here, by using radiolabeled ATP covalently bound to Arp2/3, we show that ATP is hydrolyzed on Arp2, not on Arp3, after a delay following filament branching. Hydrolysis of ATP on Arp2 promotes debranching of filaments and acts as a clock that controls the stability of dendritic actin arrays in lamellipodia. Finally, we propose that hydrolysis of ATP on G-actin in the ternary G-actin-WASP-Arp2/3 complex on branch formation destabilizes the WASP-actin interface and energetically facilitates the detachment step in the branching reaction. PMID- 12743371 TI - Exponential growth by cross-catalytic cleavage of deoxyribozymogens. AB - We have designed an autocatalytic cycle based on the highly efficient 10-23 RNA cleaving deoxyribozyme that is capable of exponential amplification of catalysis. In this system, complementary 10-23 variants were inactivated by circularization, creating deoxyribozymogens. Upon linearization, the enzymes can act on their complements, creating a cascade in which linearized species accumulate exponentially. Seeding the system with a pool of linear catalysts resulted not only in amplification of function but in sequence selection and represents an in vitro selection experiment conducted in the absence of any protein enzymes. PMID- 12743370 TI - Evidence for a genetic discontinuity between Neandertals and 24,000-year-old anatomically modern Europeans. AB - During the late Pleistocene, early anatomically modern humans coexisted in Europe with the anatomically archaic Neandertals for some thousand years. Under the recent variants of the multiregional model of human evolution, modern and archaic forms were different but related populations within a single evolving species, and both have contributed to the gene pool of current humans. Conversely, the Out of-Africa model considers the transition between Neandertals and anatomically modern humans as the result of a demographic replacement, and hence it predicts a genetic discontinuity between them. Following the most stringent current standards for validation of ancient DNA sequences, we typed the mtDNA hypervariable region I of two anatomically modern Homo sapiens sapiens individuals of the Cro-Magnon type dated at about 23 and 25 thousand years ago. Here we show that the mtDNAs of these individuals fall well within the range of variation of today's humans, but differ sharply from the available sequences of the chronologically closer Neandertals. This discontinuity is difficult to reconcile with the hypothesis that both Neandertals and early anatomically modern humans contributed to the current European gene pool. PMID- 12743372 TI - Biocomplexity and fisheries sustainability. AB - A classic example of a sustainable fishery is that targeting sockeye salmon in Bristol Bay, Alaska, where record catches have occurred during the last 20 years. The stock complex is an amalgamation of several hundred discrete spawning populations. Structured within lake systems, individual populations display diverse life history characteristics and local adaptations to the variation in spawning and rearing habitats. This biocomplexity has enabled the aggregate of populations to sustain its productivity despite major changes in climatic conditions affecting the freshwater and marine environments during the last century. Different geographic and life history components that were minor producers during one climatic regime have dominated during others, emphasizing that the biocomplexity of fish stocks is critical for maintaining their resilience to environmental change. PMID- 12743373 TI - Inhibition of p53-induced apoptosis without affecting expression of p53-regulated genes. AB - Using DNA microarray and clustering of expressed genes we have analyzed the mechanism of inhibition of wild-type p53-induced apoptosis by the cytokine interleukin 6 (IL-6) and the calcium mobilizer thapsigargin (TG). Clustering analysis of 1,786 genes, the expression level of which changed after activation of wild-type p53 in the absence or presence of IL-6 or TG, showed that these compounds did not cause a general inhibition of the ability of p53 to up-regulate or down-regulate gene expression. Expression of various p53 targets implicated as mediators of p53-induced apoptosis was also not affected by IL-6 or TG. These compounds thus can bypass the effect of wild-type p53 on gene expression and inhibit apoptosis. IL-6 and TG activated different p53-independent pathways of gene expression that include up-regulation of antiapoptotic genes. IL-6 and TG also activated different differentiation-associated genes. The ability of compounds such as cytokines and calcium mobilizers to inhibit p53-mediated apoptosis without generally inhibiting gene expression regulated by p53 can facilitate tumor development and tumor resistance to radiation and chemotherapy in cells that retain wild-type p53. PMID- 12743375 TI - Spectra of random graphs with given expected degrees. AB - In the study of the spectra of power-law graphs, there are basically two competing approaches. One is to prove analogues of Wigner's semicircle law, whereas the other predicts that the eigenvalues follow a power-law distribution. Although the semicircle law and the power law have nothing in common, we will show that both approaches are essentially correct if one considers the appropriate matrices. We will prove that (under certain mild conditions) the eigenvalues of the (normalized) Laplacian of a random power-law graph follow the semicircle law, whereas the spectrum of the adjacency matrix of a power-law graph obeys the power law. Our results are based on the analysis of random graphs with given expected degrees and their relations to several key invariants. Of interest are a number of (new) values for the exponent beta, where phase transitions for eigenvalue distributions occur. The spectrum distributions have direct implications to numerous graph algorithms such as, for example, randomized algorithms that involve rapidly mixing Markov chains. PMID- 12743374 TI - The phosphorylation of caveolin-2 on serines 23 and 36 modulates caveolin-1 dependent caveolae formation. AB - Caveolin-1 and -2 are the two major coat proteins found in plasma membrane caveolae of most of cell types. Here, by using adenoviral transduction of either caveolin-1 or caveolin-2 or both isoforms into cells lacking both caveolins, we demonstrate that caveolin-2 positively regulates caveolin-1-dependent caveolae formation. More importantly, we show that caveolin-2 is phosphorylated in vivo at two serine residues and that the phosphorylation of caveolin-2 is necessary for its actions as a positive regulator of caveolin-1 during organelle biogenesis in prostate cancer cells. Mutation of the primary phosphorylation sites on caveolin 2, serine 23 and 36, reduces the number of plasmalemma-attached caveolae and increases the accumulation of noncoated vesicles, but does not affect trafficking of caveolin-2, interaction with caveolin-1 or its biophysical properties. Thus, the phosphorylation of caveolin-2 is a novel mechanism to regulate the dynamics of caveolae assembly. PMID- 12743377 TI - Mate choice theory and the mode of selection in sexual populations. AB - Indirect new data imply that mate and/or gamete choice are major selective forces driving genetic change in sexual populations. The system dictates nonrandom mating, an evolutionary process requiring both revised genetic theory and new data on heritability of characters underlying Darwinian fitness. Successfully reproducing individuals represent rare selections from among vigorous, competing survivors of preadult natural selection. Nonrandom mating has correlated demographic effects: reduced effective population size, inbreeding, low gene flow, and emphasis on deme structure. Characters involved in choice behavior at reproduction appear based on quantitative trait loci. This variability serves selection for fitness within the population, having only an incidental relationship to the origin of genetically based reproductive isolation between populations. The claim that extensive hybridization experiments with Drosophila indicate that selection favors a gradual progression of "isolating mechanisms" is flawed, because intra-group random mating is assumed. Over deep time, local sexual populations are strong, independent genetic systems that use rich fields of variable polygenic components of fitness. The sexual reproduction system thus particularizes, in small subspecific populations, the genetic basis of the grand adaptive sweep of selective evolutionary change, much as Darwin proposed. PMID- 12743376 TI - Tracing the origin and history of the HIV-2 epidemic. AB - In this study we date the introduction of HIV-2 into the human population and estimate the epidemic history of HIV-2 subtype A in Guinea-Bissau, the putative geographic origin of HIV-2. The evolutionary history of the simian immunodeficiency virussooty mangabey/HIV-2 lineage was reconstructed by using available database sequences with known sampling dates, and a timescale for this history was calculated by using maximum likelihood methods. The date of the most recent common ancestor of HIV-2 subtype A strains was estimated to be 1940 +/- 16 and that of B strains was estimated to be 1945 +/- 14. In addition we used coalescent theory to estimate the past population dynamics of HIV-2 subtype A in a rural population of Guinea-Bissau. Parametric and nonparametric estimates of the effective number of infections through time were obtained for an equal sample of gag, pol, and env sequences. Our estimates of the epidemic history of HIV-2 subtype A in Guinea-Bissau show a transition from constant size to rapid exponential growth around 1955-1970. Our analysis provides evidence for a zoonotic transfer of HIV-2 during the first half of the 20th century and an epidemic initiation in Guinea-Bissau that coincides with the independence war (1963-1974), suggesting that war-related changes in sociocultural patterns had a major impact on the HIV-2 epidemic. PMID- 12743378 TI - Molecular paleontology of transposable elements in the Drosophila melanogaster genome. AB - We report here a superfamily of "cut and paste" DNA transposons called Transib. These transposons populate the Drosophila melanogaster and Anopheles gambiae genomes, use a transposase that is not similar to any known proteins, and are characterized by 5-bp target site duplications. We found that the fly genome, which was thought to be colonized by the P element <100 years ago, harbors approximately 5 million year (Myr)-old fossils of ProtoP, an ancient ancestor of the P element. We also show that Hoppel, a previously reported transposable element (TE), is a nonautonomous derivate of ProtoP. We found that the "rolling circle" Helitron transposons identified previously in plants and worms populate also insect genomes. Our results indicate that Helitrons were horizontally transferred into the fly or/and mosquito genomes. We have also identified a most abundant TE in the fly genome, DNAREP1_DM, which is an approximately 10-Myr-old footprint of a Penelope-like retrotransposon. We estimated that TEs are three times more abundant than reported previously, making up approximately 22% of the whole genome. The chromosomal and age distributions of TEs in D. melanogaster are very similar to those in Arabidopsis thaliana. Both genomes contain only relatively young TEs (<20 Myr old), constituting a main component of paracentromeric regions. PMID- 12743380 TI - (NH4)2ZrGe3O9: a new microporous zirconogermanate. AB - A new microporous zirconogermanate, diammonium zirconium trigermanate, (NH(4))(2)ZrGe(3)O(9) (FDZG-2), analogous to wadeite (K(2)ZrSi(3)O(9)), was hydrothermally synthesized using ZrO(NO(3))(2).2H(2)O as the source of zirconium and 1,4-diaminobutane as a structure-directing agent. Single-crystal X-ray diffraction analysis reveals that the framework structure is built up of cyclic trigermanate units crosslinked by ZrO(6) octahedra. The Zr atom lies at a site with -3 symmetry and the unique N atom of the ammonium ion lies at a site with threefold symmetry. Large cages are observed, with two NH(4)(+) cations in each. The structure contains intersecting six- and three-membered ring (6MR and 3MR) channels, but only the 6MR channels can accommodate the NH(4)(+) ions. PMID- 12743381 TI - Dirubidium dicadmium sulfate at 293 K. AB - The structure of Rb2Cd2(SO4)3 at room temperature has been refined using X-ray diffraction data. The structure is similar to that of other cubic langbeinites. However, the title compound differs from other type I langbeinites in that the bond-valence sums of the metal cations (in particular the Rb atoms) are practically equal to their nominal chemical valences. PMID- 12743382 TI - Two Ti-doped distrontium ruthenium tetraoxides: Sr2Ru0.93Ti0.07O4 and Sr2Ru0.81Ti0.19O4. AB - Crystals of titanium-doped distrontium ruthenium tetraoxide, Sr(2)Ru(1 x)Ti(x)O(4), with x = 0.07 and 0.19, were grown by floating-zone melting, and their structures were solved using single-crystal X-ray diffraction. Increasing Ti content leads to a distinctive systematic variation of cell parameters and interatomic distances with respect to the undoped material. PMID- 12743379 TI - Structural defects and the diagnosis of amyloidogenic propensity. AB - Disease-related amyloidogenic propensity has been unexpectedly found in proteins driven to adopt a monomeric uncomplexed state at high concentrations under near physiological conditions. This situation occasionally arises in new health treatments, such as kidney dialysis. Assuming that under such conditions a partial retention of native structure takes place, this work identifies a structural characteristic indicating amyloidogenic propensity: a high density of backbone hydrogen bonds exposed to water attack in monomeric structure. On this basis, we propose a diagnostic tool based on the identification of hydrogen bonds with a paucity of intramolecular dehydration or "wrapping." We use this predictor to identify potentially pathogenic mutations that foster amyloidogenic propensity in human prions. Such mutations either enhance the intramolecular dehydration of beta-sheet hydrogen bonds, thus stabilizing the nucleus for rearrangement into the scrapie fold, or contribute to the destabilization of the cellular form by introducing additional underwrapped hydrogen bonds. Our predictions are consistent with known disease-related mutations and lead to a cogent explanation of the pathogenic nature of specific mutations affecting the cellular prion protein structural wrapping. On the other hand, a different wrapping of a very similar fold, mouse doppel, induces a dramatically different level of amyloidogenic propensity, suggesting that the packing within the fold, and not the fold itself, contains the signal for aggregation. PMID- 12743383 TI - LiInSiO4: a new monovalent-trivalent olivine. AB - The structure of the olivine LiInSiO(4) (lithium indium silicate) is isotypic with LiScSiO(4) and MgMgSiO(4) (forsterite). The main differences between the title compound and the divalent-divalent olivines are found for the bond lengths and angles opposite common edges between the tetrahedron and the Li(+) and In(3+) ion sites. The tetrahedron shares one common edge with the Li(+) site and two common edges with the In(3+) site. The tetrahedron is distinctly distorted, as are the Li(+) and In(3+) sites. PMID- 12743384 TI - Disordered syn-anti hydrogen-bonded chains in 3-(ferrocenylcarbonyl)propionic acid. AB - The title compound, [Fe(C(5)H(5))(C(9)H(9)O(3))], has Fe-centroid distances of 1.6551 (11) and 1.6445 (11) A to the cyclopentadienyl rings. The carboxyl group forms hydrogen bonds in the extremely rare syn-anti chain motif, with O...O distances of 2.667 (3) and 2.655 (3) A. The carboxyl group and the hydrogen bonded chains are disordered. PMID- 12743385 TI - Tetra-n-butylammonium tetrakis(pentafluorobenzenethiolato-kappaS)aurate(III). AB - The title compound, (C(16)H(36)N)[Au(C(6)F(5)S)(4)], is the first example of a structurally characterized gold(III) complex with monodentate benzenethiolate ligands. The Au atom lies on a fourfold axis and the AuS(4) group has square planar geometry. The anion shows a two-dimensional linkage through pi-pi and C F...pi intermolecular interactions. PMID- 12743386 TI - Polymeric mu-bromo-mu-pyridine-3-carboxylato-kappa3O,O':N-mercury(II). AB - The asymmetric unit of the title polymeric complex, [HgBr(C(6)H(4)NO(2))](n) or HgBr(nic), contains mercury coordinated via two Br atoms [Hg-Br = 2.6528 (9) and 2.6468 (9) A], two carboxylate O atoms, which form a characteristic four-membered chelate ring [Hg-O = 2.353 (6) and 2.478 (7) A], and an N atom [Hg-N = 2.265 (5) A], in the form of a very irregular (3+2)-coordination polyhedron. The pronounced irregularity of the effective Hg (3+2)-coordination is a result of the rigid stereochemistry of the nicotinate ligand. According to the covalent and van der Waals radii criteria, the strongest bonds are Hg-Br and Hg-N. These covalent interactions form a two-dimensional polymer. The puckered planes are connected by van der Waals interactions, and there are only two intermolecular C-H...O hydrogen bonds [3.428 (10) and 3.170 (10) A]. PMID- 12743387 TI - The monoclinic polymorph of rac-5,7,7,12,12,14-hexamethyl-1,4,8,11 tetraazoniacyclotetradecane bis(hexafluorogermanate) tetrahydrate. AB - X-ray data were obtained for the monoclinic polymorph of rac-5,7,7,12,12,14 hexamethyl-1,4,8,11-tetraazoniacyclotetradecane bis(hexafluorogermanate) tetrahydrate, (C(16)H(40)N(4))[GeF(6)](2).4H(2)O. The tetraaza-macrocyclic cations lie across inversion centers in space group P2(1)/c. Water molecules and [GeF(6)](2-) anions form zigzag chains, which alternate in a three-dimensional network with the macrocyclic cations. The structure is sustained by multiple hydrogen bonds. PMID- 12743389 TI - Polymeric hexaaquahexakis(mu3-2,2'-oxydiacetato)trizinc(II)digadolinium(III) dodecahydrate. AB - A polymeric heterometallic compound, ([Gd(2)Zn(3)(C(4)H(4)O(5))(6)(H(2)O)(6)].12H(2)O)(n), comprising zinc(II) and gadolinium(III) cations bridged by carboxylate groups from oxydiacetate ligands, is presented. The Gd(III) cations lie at sites with crystallographic 32 symmetry and display a tricapped trigonal-prism arrangement, which is defined by six carboxyl and three ether O atoms. The Zn(II) cations lie at sites with imposed 2/m symmetry and are octahedrally coordinated by four carboxyl O atoms and two apical water ligands, which form strong intramolecular hydrogen bonds. Comparison is made with the previously reported isostructural homologous copper-gadolinium complex. PMID- 12743388 TI - Two new dimeric cadmium(II) and zinc(II) sulfate complexes with 2,4,6-tris(2 pyridyl)-1,3,5-triazine and 2,2:6',2"-terpyridine. AB - The structures of two new sulfate complexes are reported, namely di-mu-sulfato kappa(3)O,O':O"-bis(aqua[2,4,6-tris(2-pyridyl)-1,3,5-triazine kappa(3)N(1),N(2),N(6)]cadmium(II)) tetrahydrate, [Cd(2)(SO(4))(2)(C(16)H(12)N(6))(2)(H(2)O)(2)].4H(2)O, and di-mu-sulfato kappa(2)O:O'-bis[(2,2':6',2"-terpyridine-kappa(3)N(1),N(1'),N(1"))zinc(II)] dihydrate, [Cd(2)(SO(4))(2)(C(15)H(11)N(3))(2)].2H(2)O, the former being the first report of a Cd(tpt) complex [tpt is 2,4,6-tris(2-pyridyl)-1,3,5-triazine]. Both compounds crystallize in the space group P-1 and form centrosymmetric dimeric structures. In the cadmium complex, the metal center is heptacoordinated in the form of a pentagonal bipyramid, while in the zinc complex, the metal ion is in a fivefold environment, the coordination geometry being intermediate between square pyramidal and trigonal bipyramidal. Packing of the dimers leads to the formation of planar structures strongly linked by hydrogen bonding. PMID- 12743390 TI - [N-(Carboxylatomethyl)aspartato(3-)](ethylenediamine)cobalt(III) trihydrate. AB - The mononuclear title complex, [Co(C(6)H(6)NO(6))(C(2)H(8)N(2))].3H(2)O, contains an octahedrally coordinated Co(III) atom. The N-(carboxymethyl)aspartate moiety is coordinated as a tetradentate ligand, providing an OONO-donor set and forming two trans five-membered chelate rings and one six-membered chelate ring. A seven membered chelate ring is also formed, which consists of part of the six-membered chelate ring and part of one of the five-membered chelate rings. The crystal structure of the complex is stabilized by hydrogen bonds with three water molecules. PMID- 12743391 TI - trans-Diaquabis(thiosemicarbazido-kappa2N,S)nickel(II) dimaleate dihydrate. AB - In the title compound, [Ni(CH(5)N(3)S)(2)(H(2)O)(2)](C(4)H(3)O(4))(2).2H(2)O, the Ni atom lies on a center of symmetry and is coordinated by N and S atoms from two thiosemicarbazide ligands and the O atoms of two water molecules in a distorted octahedral geometry. In the asymmetric unit, the three components are linked together by one O-H...O and two N-H...O hydrogen bonds. The packing is built from molecular ribbons parallel to the b direction, stabilized by intramolecular hydrogen bonds, and by one N-H...S and two N-H...O intermolecular hydrogen bonds. The ribbons are further connected into columns by N-H...O interactions and then into a three-dimensional network by three O-H...O hydrogen bonds. PMID- 12743392 TI - Polymeric tris(mu2)-acetone-kappa2O:O)sodium polyiodide at 120 K. AB - In the title compound, [Na(C(3)H(6)O)(3)](n)(I(2))(n), all non-H atoms are in special positions of the space group P6(3)/mcm, with the Na atom in 2b, the I atom in 4c, the carbonyl O atom and the C atom attached to it both in 6g, and the methyl C atom in 12k. The H atoms of the rotationally disordered methyl groups are in 24l general positions but with occupancies of 0.5, because they occur in two sets related by a crystallographic mirror plane. Infinite chains are created by face-sharing octahedral Na-coordination polyhedra, with Na-O and Na...Na distances of 2.439 (5) and 3.2237 (4) A, respectively. I atoms form infinite linear chains, in which the I-atom separation is 3.2237 (4) A. PMID- 12743393 TI - Tripotassium tris(oxalato-kappa2O,O')aluminate bis(hydrogen peroxide) hydrate, the first example of a cyclic hydrogen-bonded H2O2 dimer. AB - In addition to associating into cyclic hydrogen-bonded dimers [O...O = 2.663 (1) and 2.914 (1) A], each hydrogen peroxide molecule in the title structure, K(3)[Al(C(2)O(4))(3)].1.75H(2)O(2).0.25H(2)O, hydrogen bonds to a neighbouring oxalate ligand [O...O = 2.700 (1) and 2.730 (1) A] and coordinates to two K(+) ions [K...O = 2.6620 (9)-2.8380 (7) A]. PMID- 12743394 TI - Dipotassium di-mu-isothiocyanato-kappa4N:S-bis[(N-salicylidene-DL-valinato kappa3O,N,O')cuprate(II)]. AB - The title compound, K(2)[Cu(2)(NCS)(2)(C(12)H(13)NO(3))(2)], consists of two K(+) cations and (N-salicylidene-D-valinato)copper(II) and (N-salicylidene-L valinato)copper(II) coordination units connected through three-atom thiocyanate (mu-NCS) bridges into a centrosymmetric dianion. The Cu(II) atom adopts a square pyramidal coordination, with three donor atoms of the tridentate Schiff base and one N atom of the bridging ligand (mu-NCS) in the basal plane. The axial position is occupied by the thiocyanate S atom of a symmetry-related ligand at an apical distance of 2.9332 (10) A. Coulombic interactions between six-coordinated K(+) ions and the heteroatoms of neighbouring dimeric anions leads to the formation of one-dimensional chains of molecules parallel to [010]. The superposition of the normals of the pyramidal base planes in a direction close to [001] indicates possible pi-pi interactions between neighbouring units. PMID- 12743395 TI - trans-Bis(dimethylglyoximato-kappa2N,N')(1-hexenyl)(pyridine-kappaN)cobalt(III): a cobaloxime-substituted terminal alkene that rapidly isomerizes to a cobaloxime substituted internal alkenyl complex. AB - An unusual cobaloxime-substituted terminal alkene, [Co(C(6)H(11))(C(4)H(7)N(2)O(2))(2)(C(5)H(5)N)], has been isolated and characterized by X-ray crystallography. The double bond in the alkene readily isomerizes, but the title compound could be isolated and structurally characterized at low temperature. PMID- 12743396 TI - A one-dimensional coordination polymer with metal-metal interactions: catena poly[[triaquabarium(II)]-mu-aqua-kappa2O:O-di-mu-thiosalicylato-kappa4O:O]. AB - In the title compound, [Ba(C(7)H(5)O(2)S)(2)(H(2)O)(4)](n), the Ba(II) atom lies on a mirror plane and is nine-coordinated by four bridging carboxylate O atoms of the thiosalicylate ligands, two bridging water molecules and three terminal water molecules. There is an intramolecular S-H.O hydrogen bond between the S and O atoms in the thiosalicylate ligand. A one-dimensional coordination polymer is formed via weak metal-metal interactions along polymeric zigzag chains. PMID- 12743397 TI - Bis(O-ethyl dithiocarbonato-kappa2S,S')palladium(II). AB - The Pd atom in the title compound, [Pd(C(3)H(5)OS(2))(2)], lies on an inversion center and adopts a square-planar coordination geometry defined by the four S atoms of the two dithiocarbonate (xanthate) ligands. In the solid state, the molecules aggregate into layers in which the rows of molecules alternate their orientation to allow each Pd atom to interact with two symmetry-equivalent S atoms of the xanthate ligands of adjacent molecules, generating a pseudo octahedral environment around each Pd atom. This weak interaction of 3.3579 (7) A can be classified as a closed-shell electrostatic intermolecular interaction. PMID- 12743399 TI - Head-to-head dimers in the zwitterion of 1-hydroxy-3-(pyrrolidin-1-yl)propylidene 1,1-bisphosphonic acid (EB 1053). AB - The title compound, 1-hydroxy-1-phosphono-3-(1-pyrrolidinio)propylidene-1 phosphonate, C(7)H(17)NO(7)P(2), is a member of the bisphosphonate class of drugs. As a zwitterion, it possesses a negative charge on one of the PO(3) groups and a positive charge on the pyrrolidine N atom. A zwitterion makes a contact with a neighbouring ion through the hydroxyl O atom and two phosphonyl O atoms, one each from two different PO(3) groups. Hydrogen bonding involves O-H.O and N H.O interactions; the former are involved in the formation of head-to-head dimers, while the latter join the dimers into a chain running along the crystallographic b axis. PMID- 12743398 TI - (4S,5S)-4-[(1R)-1,2-Dihydroxyethyl]-5-tridecyl-1,3-oxazolidin-2-one. AB - The title compound, C(18)H(35)NO(4), is a new bioactive amphiphilic lipid with a cis-substituted 1,3-oxazolidin-2-one head group. In the crystal structure, the molecules form intercalating bilayers in which the oxazolidinone head groups are joined together by hydrogen bonds into chains. PMID- 12743400 TI - (+/-)-5-(4-methoxyphenylaminocarbonyl)-1-azabicyclo[3.3.0]octan-2-one benzene solvate. AB - The title trans prolyl amide exists as a benzene solvate, C(15)H(18)N(2)O(3).C(6)H(6), with positional disorder of the prolyl ring. The molecular structure is influenced by a close intramolecular N-H...N contact that provides structural support for the intramolecular catalysis of peptidyl-prolyl cis-trans isomerization. PMID- 12743401 TI - 4-[(3-chlorophenyl)diazenyl]-2 ([tris(hydroxymethyl)methyl]aminomethylene)cyclohexa-3,5-dien-1(2H)-one. AB - The title compound, C(17)H(18)ClN(3)O(4), adopts the keto-amine tautomeric form and displays an intramolecular N-H...O hydrogen bond [N...O = 2.639 (2) A]. The configuration around the azo N=N double bond is trans, and the dihedral angle between the planes of the two aromatic rings is 20.5 (2) degrees. The molecules are linked by O-H...O hydrogen bonds to form a three-dimensional network. PMID- 12743402 TI - 6-amino-2-(3,4-dimethoxybenzylamino)-3-methyl-5-nitrosopyrimidin-4(3H)-one: hydrogen-bonded sheets form interdigitated bilayers. AB - Molecules of the title compound, C(14)H(17)N(5)O(4), exhibit a highly polarized molecular-electronic structure. The molecules are linked into sheets by two N H...O hydrogen bonds [H...O = 2.03 and 2.02 A, N...O = 2.836 (2) and 2.887 (2) A, and N-H...O = 153 and 168 degrees ], augmented by a single C-H...O hydrogen bond [H...O = 2.47 A, C.O = 3.403 (2) A and C-H...O = 166 degrees ]. Pairs of sheets, related by inversion, form bilayers with interdigitated dimethoxybenzyl substituents, weakly linked by a further C-H.O interaction [H...O = 2.50 A, C...O = 3.350 (2) A and C-H...O = 146 degrees]. PMID- 12743403 TI - (R)-1-Phenylethylammonium 6-amino-5-nitropyrimidine-2,4(1H,3H)-dionate (R)-1 phenylethylamine hemisolvate: hydrogen-bonded sheets of ions with pendent solvent molecules. AB - In the title compound, 2C(8)H(12)N(+).2C(4)H(3)N(4)O(4)(-).C(8)H(11)N, the anions are linked by paired N-H...N hydrogen bonds [H.N = 2.07 and 2.11 A, N.N = 2.942 (3) and 2.978 (3) A and N-H.N = 173 and 170 degrees] and by paired N-H...O hydrogen bonds [H.O = 1.98 and 2.05 A, N...O = 2.855 (3) and 2.917 (3) A, and N H...O = 173 and 167 degrees ] into chains of rings. These chains are linked into sheets by further N-H...O hydrogen bonds in which all of the donors are provided by the cations [H....O = 1.83-2.17 A, N...O = 2.747 (3)-2.965 (3) A and N-H...O = 141-168 degrees]. The neutral amine molecule is pendent from the sheet and is linked to it by a single N-H...N hydrogen bond [H...N = 2.00 A, N...N = 2.901 (3) A and N-H...N = 175 degrees]. PMID- 12743404 TI - Hydrogen bonding in 1-carbamoylguanidinium methylphosphonate monohydrate. AB - The title compound, C(2)H(7)N(4)O(+).CH(4)O(3)P(-).H(2)O, crystallized with one carbamoylguanidinium cation, one methylphosphonate anion and one water molecule in the asymmetric unit. All H atoms of the carbamoylguanidinium ion are involved in a hydrogen-bonded network. The CH(3)PO(2)(OH) anions, together with the water molecules, build O-H...O hydrogen-bonded ribbons around a 2(1) screw axis parallel to the b axis. Neighbouring ribbons are not directly connected via hydrogen bonding. The carbamoylguanidinium cations are linked to these ribbons by N-H...O bridges and build a slightly buckled layer structure, the interlayer distance being b/2. PMID- 12743405 TI - The host-guest complex between cone-25,26:27,28-bis(methylenedioxy)calix[4]arene and dichloromethane. AB - The title compound, 25,26:27,28 bis(methylenedioxy)pentacyclo[19.3.1.1(3,7).1(9,13).1(15,19)]octacosa 1(25)3,5,7(28),9,11,13(27),15,17,19(26),21,23-dodecaene dichloromethane solvate, C(30)H(24)O(4).CH(2)Cl(2), possesses crystallographic twofold symmetry in both components. The calixarene shows a pinched cone conformation with an elliptical cavity, in which the guest dichloromethane solvent molecule is accommodated. The contact distance between guest and host (H.ring centroid = 2.44 A) is extremely short. PMID- 12743407 TI - The sesquiterpenoid nootkatone and the absolute configuration of a dibromo derivative. AB - Nootkatone, or (4R,4aS,6R)-4,4a,5,6,7,8-hexahydro-4,4a-dimethyl-6-(1 methylethenyl)naphthalen-2(3H)-one, C(15)H(22)O, a sesquiterpene with strong repellent properties against Formosan subterranean termites and other insects, has the valencene skeleton. The dibromo derivative (1S,3R,4S,4aS,6R,8aR)-1,3 dibromo-6-isopropyl-4,4a-dimethyl-1,2,3,4,5,6,7,8-octahydronaphthalen-2-one, C(15)H(24)Br(2)O, has two independent molecules in the asymmetric unit, which differ in the rotation of the isopropyl group with respect to the main skeleton. The C-Br distances are in the range 1.950 (4)-1.960 (4) A. Both independent molecules form zigzag chains, with very short intermolecular carbonyl-carbonyl interactions, having the perpendicular motif and O...C distances of 2.886 (6) and 2.898 (6) A. These chains are flanked by intermolecular Br...Br interactions of distances in the range 4.067 (1)-4.218 (1) A. The absolute configuration of the dibromo derivative was determined, from which that of nootkatone was inferred. PMID- 12743408 TI - A "tetrameric" 3,4-di-p-tolyl-1,2,5-oxadiborole derivative. AB - In the title compound, 1,1,6a,7,9a,10-hexachloro-2,3,5,6,8,9,11,12-octa-p-tolyl 1,6a,9a,12a-tetraborata-3a,4a,7,10-tetrabora-4a(1),6b,9b,12b-tetraoxonia-4 oxatetracyclopenta[1,2-a:2,1,5-de:1,2-g:1,2-i]naphthalene dichloromethane pentasolvate, C(64)H(56)B(8)Cl(6)O(5).5CH(2)Cl(2), two condensed oxadiborole rings are attached to two further oxadiborole rings in a type of donor-acceptor bonding, thus forming a ten-membered alternating (B-O)(5) naphthalene-like arrangement as the central building block. PMID- 12743406 TI - 1,3-thiazolidine derivatives from regioselective [2+3]-cycloadditions of azomethine ylides with thioketones. AB - The title compounds, namely dimethyl (2RS)-2,3-diphenyl-1,3-thiazolidine-5-spiro 2'-adamantane-4,4-dicarboxylate methanol solvate, C(28)H(31)NO(4)S.0.275CH(4)O, and dimethyl (4RS)-3,4-diphenyl-1,3-thiazolidine-5-spiro-9'-(9'H-fluorene)-2,2 dicarboxylate, C(31)H(25)NO(4)S, were obtained from dipolar [2+3]-cycloadditions of an azomethine ylide with adamantanethione and thiofluorenone, respectively. The structures show that the choice of thioketone affects the regioselectivity of the cycloaddition. The asymmetric unit of the former structure contains two molecules of the thiazolidine derivative plus a site for a partial occupancy (55%) methanol molecule. O-H...O and C-H...O interactions link two of each of these entities into closed centrosymmetric hexamers. The five-membered ring in each structure has an envelope conformation. PMID- 12743409 TI - Two hydrates of 2,6-bis(1H-benzimidazol-2-yl)pyridine. AB - The structures of the mono- and sesquihydrates of 2,6-bis(1H-benzimidazol-2 yl)pyridine (bbip) are reported. Phase (I), C(19)H(13)N(5).H(2)O, has one water and one bbip molecule in the asymmetric unit, while phase (II), C(19)H(13)N(5).1.5H(2)O, has three water molecules and two bbip molecules in the asymmetric unit. The compounds exhibit very similar molecular geometries but different packing organizations, which result from intricate hydrogen-bonding schemes. PMID- 12743410 TI - Five (1H-pyrrol-2-yl)pyridines. AB - The title (1H-pyrrol-2-yl)pyridines, C(9)H(8)N(2), substituted at the ortho, meta, and para positions of the pyridine ring all have hydrogen-bonded arrangements with geometrically similar, nearly linear, N(pyrrole) H...N(pyridine) hydrogen bonds of average length. The graph sets for the ortho, meta, and three para polymorphs are R(2)(2)(10), C(6), C(7), C(7), and R(4)(4)(28), respectively. PMID- 12743412 TI - [Et4N][7-Me2S-nido-B11H12]. AB - Tetraethylammonium 7-dimethylsulfanyl-nido-dodecahydroundecaborate, [Et(4)N][7 Me(2)S-nido-B(11)H(12)] or C(8)H(20)N(+).C(2)H(18)B(11)S(-), is a product of the deprotonation of [7-Me(2)S-nido-B(11)H(13)] with KHBEt(3) and precipitation with tetraethylammonium chloride. The effect of removing one endo-terminal H atom is to cause a general contraction of the open-face B-B distances. PMID- 12743411 TI - meso-Bis(phenylsulfinyl)methane. AB - The title compound, meso-C(13)H(12)O(2)S(2), is in an anti conformation, with R and S configurations around the S atoms. The two O atoms are trans to each other, and the same applies for the two benzene rings. The phenylsulfinyl groups are nearly orthogonal to the central dithiomethane group, and the orientation of the two phenyl rings are determined by the interactions in which they are involved. The packing is built from molecular columns stabilized by weak C-H...O interactions. PMID- 12743413 TI - 9alpha-Fluoro-16alpha-methyl-3,11-dioxoandrosta-1,4-diene-17beta-carboxylic acid: catemeric hydrogen bonding and acetic acid solvation in a steroidal keto acid related to dexamethasone. AB - The title keto acid crystallizes as a solvate, C(21)H(25)FO(4).C(2)H(4)O(2), with two molecules each of steroid and acetic acid per asymmetric unit. The former are approximately parallel, with opposite end-to-end orientation, and form translational carboxyl-to-ketone hydrogen-bonding catemers [O...O = 2.679 (6) and 2.650 (5) A, and O-H...O = 165 and 162 degrees] that involve the 3-ketone group and follow the a axis. The acetic acid molecules are paired by hydrogen bonding, and neither they nor the F atom nor the 11-ketone group play any overt role in the hydrogen-bonding scheme of the steroid. Intermolecular C-H...O=C close contacts involving three different neighboring molecules exist to the 11-ketone group, the steroidal carboxyl group and one of the acetic acid molecules. PMID- 12743414 TI - Two polymorphs of bis(2-bromophenyl) disulfide. AB - Colourless crystals of the title compound, bis(2-bromophenyl) disulfide, C(12)H(8)Br(2)S(2), are obtained from the reaction of 2-bromophenylmercaptan with metallic sodium and either zinc chloride or cadmium chloride in methanol. In the presence of Zn(II) ions, the crystals are orthorhombic (space group Pbca, with Z' = 1); with Cd(II) ions present, the product is triclinic (space group (-)P1, with Z' = 4). Both polymorphs exhibit significant intramolecular C-H.S hydrogen bonds. In the orthorhombic form, molecules are linked by intermolecular C-H...Br hydrogen bonds, while in the triclinic form, molecules exhibit Br...Br contacts. PMID- 12743415 TI - Dipropyl 3,6-diphenyl-1,2-dihydro-1,2,4,5-tetrazine-1,2-dicarboxylate. AB - The title compound, C(22)H(24)N(4)O(4), was prepared from propyl chloroformate and 3,6-diphenyl-1,2-dihydro-s-tetrazine. This reaction yields the title compound rather than dipropyl 3,6-diphenyl-1,4-dihydro-s-tetrazine-1,4-dicarboxylate. The 2,3-diazabutadiene group in the central six-membered ring is not planar; the C=N double-bond length is 1.285 (2) A, and the average N-N single-bond length is 1.401 (3) A, indicating a lack of conjugation. The ring has a twist conformation, in which adjacent N atoms lie +/- 0.3268 (17) A from the plane of the ring. The molecule has twofold crystallographic symmetry. PMID- 12743416 TI - Helicobacter pylori and pancreatic disease. PMID- 12743417 TI - Effects of the imidazoline binding site ligands, idazoxan and efaroxan, on the viability of insulin-secreting BRIN-BD11 cells. AB - CONTEXT: Certain imidazoline drugs stimulate insulin secretion acutely but their longer term effects on the viability of pancreatic beta-cells are less well characterised. Indeed, some reports have suggested that imidazolines can be toxic to beta-cells while others have reported protective effects against other cytotoxic agents. OBJECTIVE: In order to address these discrepancies, the effects of two structurally related imidazolines, efaroxan and idazoxan, on the viability of clonal BRIN-BD11 beta-cells, were compared. DESIGN AND MAIN OUTCOME MEASURES: BRIN-BD11 cells were exposed to test reagents and their viability monitored by measuring cellular reducing ability and DNA fragmentation. Nitric oxide was measured indirectly via medium nitrite formation. RESULTS: Efaroxan (up to 100 micro M) did not directly affect BRIN-BD11 cell viability in the absence of other agents and it did not protect these cells against the cytotoxic effects of interleukin-1beta. Indeed, analysis of DNA fragmentation in BRIN-BD11 cells revealed that efaroxan enhanced the level of damage caused by interleukin-1beta. Idazoxan caused a time- and dose-dependent loss of BRIN-BD11 cell viability in the absence of other ligands. This was associated with marked DNA degradation but was not associated with formation of nitric oxide. The effects of idazoxan were insensitive to blockade of alpha(2)-adrenoceptors or 5-HT(1A) (5 hydroxytryptamine; serotonin) receptors. CONCLUSIONS: The results confirm that idazoxan is cytotoxic to beta-cells but show that efaroxan is better tolerated. However, since efaroxan enhanced the cytotoxic effects of interleukin-1beta, it appears that this imidazoline may sensitise BRIN-BD11 cells to the damaging effects of certain cytokines. PMID- 12743418 TI - Are glands present in goose pancreatic ducts? A light microscope study. AB - OBJECTIVE: The objective of the present study was to investigate the histological structure of goose pancreatic ducts. DESIGN: Tissue samples from the lobes and ducts of the pancreas were dissected under deep ether anesthesia. INTERVENTIONS: Sections were stained using Crossmon's connective tissue method for general observations and Gomori's method for pancreatic islet cells. RESULTS: The glands were found intermittently inside the connective tissue of the ducts starting from the interlobular ducts to the point where the pancreas emptied its contents into the duodenum as well as inside the muscular layer of the pancreatic ducts. Those glands contained centro-acinar cells and also had the same staining features as the acinus. CONCLUSIONS: To our knowledge, this was the first report of the presence of glands in the ducts of goose pancreas. PMID- 12743419 TI - Acute pancreatitis due to pravastatin therapy. AB - CONTEXT: Few data exist about the incidence of drug-induced pancreatitis in the general population. Drugs are related to the etiology of pancreatitis in about 1.4-2% of cases. Statins are generally well tolerated. Acute pancreatitis has been reported in a few cases treated with atorvastatin, fluvastatin, lovastatin and simvastatin. CASE REPORT: We report the case of a 56-year-old patient who, after 6 months of treatment with pravastatin 20 mg once daily for hypercholesterolemia, presented with acute pancreatitis. Other causes of the disease were ruled out. Five months later, the patient, on his own initiative, reintroduced pravastatin and acute pancreatitis recurred after 3 days. CONCLUSION: To our knowledge this is the first report of pravastatin-induced pancreatitis and further strengthens the fact that statins may cause acute pancreatitis. PMID- 12743420 TI - New approaches for the detection and characterization of alpha-fetoprotein epitope variants. AB - Alpha-fetoprotein (AFP) is an embryo-specific protein of mammalian sera that consists of a single glycosylated polypeptide chain (approximately 70 kD) with 5 6 epitope clusters located on the native molecule. We have elaborated three approaches for the separation of AFP into fractions (variants) that are different in the distinct epitope expression on the native molecule. For this, we have used three technologies largely based on immunoaffinity electrochromatography and electrophoresis/immunoblotting. The separation of these variants was facilitated using monoclonal antibodies previously characterized to the 5 epitope clusters and two individual epitopes on AFP in the ISOBM international workshop. We have shown that these approaches facilitate the identification of cryptic epitopes in AFP, and the relevance of these findings is discussed. PMID- 12743421 TI - Collagenases (MMP-1, -8 and -13) and trypsinogen-2 in fluid from benign and malignant ovarian cysts. AB - Proteolytic enzymes, such as matrix metalloproteinases (MMPs) and tumor associated trypsinogens (TAT), play a pivotal role in tumor invasion and metastasis. Among MMPs, the interstitial collagenases (MMP-1, -8 and -13) can initiate collagenolysis. In this study, we have studied the levels of MMP-1, -8 and -13 in relation to the level of trypsinogen-2 in fluid from benign and malignant ovarian cysts. Elevated MMP-8 levels occur in many ovarian cyst fluids, and high MMP-8 levels are associated with malignancy. The concentrations of trypsinogen-2 correlate with those of MMP-8, but it remains to be shown whether trypsin-2 plays a role as its activator in vivo. The strong expression of MMP-8 over MMP-1 and MMP-13 in malignant ovarian tumors may indicate that MMP-8 participates in the protease cascades associated with the invasiveness of ovarian tumors. PMID- 12743422 TI - The expression of hCG receptor mRNA in four human ovarian cancer cell lines varies considerably under different experimental conditions. AB - In the present study, expression and regulation of hCG receptor mRNA were analyzed in four established human ovarian cancer cell lines using different concentrations of hCG, EGF, and 8-bromo-cAMP for different periods between 6 and 72 h. The cells were examined for the hCG receptor using the reverse transcriptase polymerase chain reaction with specific primers amplifying the hCG receptor gene. Receptor mRNA was found in all cell lines. In the line OVCAR-3, it was expressed in all samples independent of kind and concentration of the receptor agonist and incubation period. In the line COLO-704, the hCG receptor gene was expressed only in unstimulated samples, but not in the samples incubated with a receptor agonist. The cell line EFO-21 showed a downregulation of receptor mRNA after 24 h of treatment with 25 IU/ml hCG and after 6 h of treatment with 250 IU/ml hCG or 100 ng/ml EGF. The mRNA reappeared within 24-48 h. The cell line EFO-27 showed a downregulation of receptor mRNA after 6 h of incubation with 250 IU/ml hCG. Agarose gel electrophoresis and sequencing of the polymerase chain reaction products revealed four cDNA fragments resulting from an alternative splicing of the primary transcript. The results of the study demonstrate that the expression of hCG receptor mRNA in ovarian cancer cell lines varies considerably under different experimental conditions. We showed that ovarian cancer cells can produce hCG receptors when needed or wanted. The inherent mechanisms which rule this phenomenon need further evaluation. PMID- 12743423 TI - Expression of bax and p53 proteins in the tumorigenesis and progression of breast carcinomas. AB - OBJECTIVES: Dysregulation of normal programmed cell death mechanisms plays an important role in the pathogenesis of breast cancer. The purpose of this study was to investigate the role of bax and p53 expression in tumorigenesis and progression of breast carcinoma as well as their relationship with proliferative and apoptotic activity. METHODS: We used immunohistochemical methods and in situ detection of apoptotic cells to examine 30 carcinomas in situ (CIS), 131 invasive breast carcinomas and 45 lymph node metastases. RESULTS: In 25% (33 of 131) of invasive breast carcinomas examined, bax expression was absent, while p53 accumulation was present in 37% (49 of 131). Interestingly, p53 accumulation and loss of bax expression occur in breast CIS as frequently as in invasive breast carcinoma. Thus, in 17% (5 of 30) of CIS bax expression was absent, and 30% (9 of 30) presented nuclear expression of p53. p53 accumulation was related to apoptosis and proliferative activity. However, the protein level of bax was unrelated to all parameters studied, including proliferation and apoptosis of tumor cells. A multivariate analysis of disease-free survival demonstrated that p53 accumulation and bax expression are not significant independent indicators of prognosis in operable breast carcinoma. Our results also show that the proportion of bax- and p53-positive cells does not vary between primary and metastatic tumors. CONCLUSIONS: p53 accumulation and loss of bax expression influence the acquisition of a malignant phenotype but seem to have no further impact on tumor progression. PMID- 12743425 TI - Survivin gene expression positively correlates with proliferative activity of cancer cells in esophageal cancer. AB - OBJECTIVE: The correlation between survivin gene expression and the occurrence of spontaneous apoptosis, proliferative activity of cancer cells, tumor angiogenesis, and abnormal p53 nuclear accumulation was evaluated in esophageal cancer. METHODS: A total of 57 patients, on whom surgical esophageal resection had been performed between 1993 and 2000, were enrolled in this study. Total RNA was extracted from tumors and noncancerous epithelia. Expression levels of survivin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) messenger RNA (mRNA) were analyzed quantitatively by real-time reverse transcriptase polymerase chain reaction (quantitative PCR). The proliferative activity of cancer cells, apoptotic cancer cells, microvessel density, and abnormal p53 nuclear accumulation were analyzed in these tumors. RESULTS: The expression level of tumor survivin mRNA described as survivin/GAPDH (s/G) ratio was higher than that of noncancerous tissue (p = 0.0003), but did not correlate with lymph node metastasis, with the depth of tumor invasion, with the occurrence of apoptosis or with tumor angiogenesis. However, the tumor s/G ratio positively correlated with the proliferative activity and p53 nuclear accumulation in cancer cells. The 5 year survival rate of 53 patients was 37% (we excluded 4 patients who died from operative complications from this survival study). The 5-year survival rate of 27 patients with a high tumor s/G ratio (28.6%) was lower than that of 26 patients with a low tumor s/G ratio (46.2%, p = 0.041). High tumor s/G ratio was detected as an important prognostic factor independent of tumor stage. CONCLUSION: Detection of survivin mRNA by quantitative PCR provided us with important prognostic and biological information regarding esophageal cancer patients. PMID- 12743424 TI - Colonic tumor CEA, CSAp and MUC-1 expression following radioimmunotherapy or chemotherapy. AB - Understanding the changes in tumor biology following cytotoxic therapy may lead to a better understanding of the properties of surviving tumor cell populations and to an improved ability to target and treat these cells. This report addressed the time-dependent dynamic alterations in the expression of three tumor associated antigens: carcinoembryonic antigen (CEA), colon-specific antigen (CSAp) and mucin-1 (MUC-1) following chemotherapy with 5-fluorouracil (5-FU) or radioimmunotherapy (RAIT; (131)I-labeled anti-CEA IgG) in human colonic tumor xenografts. Immunoassay results show that CEA and MUC-1 expression all increase rapidly after either 5-FU or RAIT. GW-39 tumors show a 2.7-fold increase in CEA expression after a maximum tolerated dose of RAIT, being highest after 21 days, while LS174T and HT-29 tumors maximally increase expression 8.3- and 2.6-fold on day 7 after RAIT, respectively. The change in LS174T is short-term, whereas the change in HT-29 is maintained for at least 4 weeks. Serum CEA levels in these tumor- bearing mice also increase in parallel to the changes observed in tumor. MUC-1 increases 2.5-fold by day 5-7 following RAIT in LS174T tumors and 6-fold by day 14 following RAIT in GW-39 tumors, with a corresponding increase in serum MUC 1. Dramatic increases in CSAp after RAIT were also demonstrated in GW-39 tissue by immunohistochemistry. Thus, these data indicate that the response of tumor cells to low-dose-rate radiation from RAIT or to chemotherapy is associated with an increase of CEA, MUC-1 and CSAp. PMID- 12743426 TI - Rearrangement of squamous cell carcinoma antigen genes--detection of SCCA fusion transcripts. AB - Squamous cell carcinoma antigen (SCCA), a member of the serine protease inhibitor (serpin) family, is a tumor-associated antigen and a serological marker for squamous cell carcinomas (SCC). Elevated serum levels are correlated with the clinical stage of the disease. Gene cloning has previously revealed two tandemly arrayed genes, SCCA1 and SCCA2. Using RT-PCR, an SCCA1/A2 transcript was identified in 6 out of 8 cell lines and the reciprocal SCCA2/A1 transcript was identified in 6 out of 8 analysed cell lines. Southern blot analysis showed an aberrant band pattern in 3 out of 5 cell lines. The cell lines demonstrating a normal band pattern corresponded to the cell lines where no fusion transcript was detected using PCR. Complex binding studies show that SCCA1/A2 binds to cathepsin G but not to cathepsin L, indicating that the SCCA1/A2 fusion protein has the specificity of SCCA2, but the transcription may be regulated as that of SCCA1. SCCA2/A1 reacts in the opposite way. The clinical relevance of these fusion transcripts is not yet known. Studies using primary tumours are underway to elucidate if these fusion transcripts are tumour specific and if they might be used as a diagnostic and prognostic marker for SCC. PMID- 12743427 TI - Serum soluble CD27, but not thymidine kinase, is an independent prognostic factor for outcome in indolent non-Hodgkin's lymphoma. AB - Non-Hodgkin's lymphoma (NHL) forms a heterogeneous group of diseases. Tumor markers may help to identify high-risk patients who might benefit from more aggressive therapy. Serum soluble CD27 (sCD27) and thymidine kinase (TK) are potentially valuable markers, since sCD27 has previously been shown to be related to tumor load and TK to proliferation of malignant cells. We determined serum sCD27, TK, beta-2-microglobulin (beta(2)M) and lactic dehydrogenase (LD) levels at diagnosis in 79 lymphoma patients and correlated these parameters with the stage of disease, the International Prognostic Index (IPI) score and survival. Receiver operator characteristic (ROC) curve analysis showed an excellent ability for sCD27 to discriminate between low- and high-stage disease (p < 0.001), especially in indolent lymphomas. No discriminative value for TK, beta(2)M or LD was found. For aggressive NHL, sCD27, TK, beta(2)M and LD did predict survival in the univariate analyses. However, LD was found to be the most independent prognostic factor in a multivariate Cox regression model. In indolent lymphomas, sCD27 proved to be a powerful marker to predict progression-free survival (p = 0.008). Taken together, the results of the ROC curve and survival analysis suggest that substitution of LD by sCD27 in the IPI may be considered for indolent lymphomas to enhance the prognostic value. A study in a larger cohort of patients is required to validate this approach. PMID- 12743428 TI - Evaluation of geometric and dosimetric inaccuracies of stereotactic irradiation in the rat brain. AB - OBJECTIVE: To evaluate geometric and dosimetric inaccuracies in the irradiation of the rat brain with the Leksell Gamma Knife. MATERIALS AND METHODS: Altogether three types of dosimeters were employed for these measurements: (a) a thermoluminescent dosimeter, (b) a semiconductor detector and (c) a polymer gel dosimeter. The thermoluminescent dosimeter and the semiconductor detector were calibrated using an ion chamber and then implanted in the brain of a rat cadaver and used for absolute dose determination. A special glass phantom mimicking exactly the shape of the rat body filled with the polymer gel was used for measurements of the relative dose distribution and evaluation of geometric inaccuracies during the stereotactic irradiation in the rat brain. RESULTS: Both thermoluminescent and semiconductor detectors, due to their size, measured mean doses. The observed results demonstrated that the Leksell GammaPlan can be employed for the calculation of absorbed doses in irradiation of experimental animals. In our case, it was necessary to apply a correction factor of 1.078 for the absolute absorbed dose to obtain reliable results. A comparison of calculated dose profiles using the treatment planning system in all three axes with those measured by the polymer gel dosimeter demonstrated a very good geometric agreement with the mean deviation in profile position of 0.5 mm. CONCLUSION: The results indicate that this technique can effectively check the geometric and dosimetric accuracy of stereotactic irradiation in the rat brain. The Leksell GammaPlan can be employed for the calculation of absorbed doses, but the correction factor of 1.078 had to be applied for the absolute dose calculations in our irradiation geometry. PMID- 12743429 TI - The effects of posteroventral pallidotomy on balance function in patients with Parkinson's disease. AB - Parkinson's disease is a chronic, progressive neurodegenerative disorder resulting from dopaminergic cell loss in the pars compacta of the substantia nigra. Conventional treatment of Parkinson's disease consists of pharmacological replacement of dopamine. A treatment alternative, posteroventral pallidotomy (PVP), has been used for medically intractable stages of the disease. The purpose of this study was to evaluate the effects of PVP on balance function, as measured by dynamic posturography, in patients with medically intractable Parkinson's disease. Five subjects were studied within 2 days prior to and within 6 months following PVP. Pretreatment abnormalities were found in vestibular, visual, and somatosensory processing in balance function. Posteroventral pallidotomy resulted in improvement in vestibular compensation of posture in some patients, which may be at least partially due to an improvement in latencies to respond to changes in stance. Dynamic posturography is an effective tool in the evaluation of balance and posture in patients with advanced Parkinson's disease. PMID- 12743430 TI - Intractable epilepsy and olfactory bulb hamartoma. A case report. AB - The objective of this case report is to demonstrate that a benign tumour arising solely from the olfactory bulb can act as a primary epileptic focus and to illustrate the difficulty in making a preoperative diagnosis of pathological lesions in this area. The case of a 29-year-old male with intractable epilepsy is presented. Radiological imaging demonstrated a conspicuous, calcified lesion in the mesial inferior frontal lobe. Electroclinical findings confirmed this to be the epileptogenic zone. Surgery and subsequent histology surprisingly revealed this focal lesion to be a benign hamartoma arising solely from the olfactory bulb. Resection resulted in seizure resolution. Tumours of the olfactory apparatus are extremely rare, but can present with epilepsy. PMID- 12743431 TI - Experimental trigeminal glycerol injection in dogs: histopathological evaluation by light and electron microscopy. AB - We investigated the effects of percutaneous gasserian glycerol injection in dogs and reviewed the histopathological changes. Experiments were performed in 16 adult healthy mongrel dogs. In group 1 (8 dogs) normal saline and in group 2 (8 dogs) pure glycerol was injected in the right trigeminal ganglion. After these procedures, dogs in each group were sacrificed after 24 h (3 dogs), 7 days (3 dogs), 21 days (2 dogs). The trigeminal ganglion and nerve of both sides were removed by using microsurgical techniques and examined by light and electron microscopy. Group 1: in all sections, nerve cells, myelinated and nonmyelinated fibers revealed normal patterns with slight fibrosis. Group 2: in all sections, myelinated fibers showed disintegration and swelling of the myelin sheath, rupture of axon continuity, destruction of basal lamina, deformation of the myelin-axon relationship by both light microscopy and electron microscopy. The sections examined by electron microscopy also showed axonolysis in nonmyelinated fibers. The changes after 7 and 21 days were less prominent than after 24 h. In the left sides, there are no pathological changes. Glycerol has a neurolytic effect on the dog's trigeminal ganglion. These effects were not specific and selective for myelinated and nonmyelinated nerve fibers. PMID- 12743432 TI - Stereotactic radiosurgery for type 2 neurofibromatosis acoustic neuromas: patient selection and tumour size. AB - Acoustic neuromas which are secondary to type 2 neurofibromatosis (NF2) respond less well to radiosurgery than unilateral sporadic disease. To refine the selection of these patients, a regression analysis was performed examining the response to radiosurgery of 114 NF2 tumours. The major determinant of outcome was tumour volume (p < 0.001). Calculating sensitivity and specificity values for different tumour volume limits gives a sensitivity value of 0.96 for a volume limit of 10 cm(3). This suggests that the size constraints that apply to the radiosurgical management of NF2 acoustic neuromas differ and are more restricted than those which are accepted for acoustic neuromas in general. PMID- 12743433 TI - Lansoprazole fast disintegrating tablet: a new formulation for an established proton pump inhibitor. AB - Lansoprazole is a proton pump inhibitor (PPI) which is an effective and well tolerated treatment option in the management of acid-related disorders. Lansoprazole fast disintegrating tablet (LFDT)--a new, patient-friendly and more convenient formulation of lansoprazole which can be taken with or without water- is the first PPI to be made available as an orally disintegrating tablet. It represents an innovative drug delivery system, comprising enteric-coated microgranules of lansoprazole compressed with an inactive, rapidly dispersing matrix to form a tablet. When the tablet is placed on the tongue and sucked gently it disintegrates rapidly in the mouth, releasing the enteric-coated microgranules which are swallowed with the patient's saliva without water. Alternatively, the tablet can be swallowed with a drink of water. Studies have shown that the bioavailability of LFDT is comparable to lansoprazole capsules, at both 15 and 30 mg doses; the indications and recommended dosages for LFDT are therefore identical to lansoprazole capsules. The new formulation may be of particular benefit to those with active life-styles who do not always have water available, patients who have difficulty in swallowing, and elderly patients. PMID- 12743435 TI - Gastrointestinal dysfunction in a community sample of subjects with symptoms of irritable bowel syndrome. AB - BACKGROUND/AIM: Irritable bowel syndrome (IBS) affects about 15-20% of the population of the Western countries. Traditionally, IBS has been an exclusion diagnosis, but recently definitions have emerged from population-based research. The aim of this population-based study was to evaluate any association between gastrointestinal pathophysiology and IBS in subjects with symptoms of IBS compared to subjects with no abdominal complaints. METHODS: From a random sample of 2,656 participants, subjects with IBS (32) together with subjects without abdominal complaints (26), were invited for further evaluation. IBS was defined as more than weekly experience of abdominal pain and distension, and in addition either borborygmia or altering stool consistency. The diagnostic work-up consisted of gastroscopy, manometry and 23-hour pH and pressure recordings of the oesophagus, lactose tolerance test, barium enema, measurement of colonic transit time, and rectoscopy. RESULTS: Compared to the group without abdominal complaints significantly more subjects with IBS had spasms of the colon (OR = 10.2 (1.2 87.3)), and abnormal contractions of the oesophagus at manometry (OR = 9.1 (1.1 78.2)). Furthermore, there was a non-significant tendency towards spasms at 23 hour pH and pressure recordings (OR = 3.58 (0.4-35.2)), and more discomfort at lactose tolerance test (OR = 5.8 (0.6-51.3)) in persons with IBS compared to subjects without abdominal complaints. CONCLUSION: The results of this population based study indicate that signs of gastrointestinal dysmotility and hyperperception are more prevalent in subjects with IBS than in subjects without abdominal complaints. PMID- 12743434 TI - Comparable efficacy of pantoprazole and omeprazole in patients with moderate to severe reflux esophagitis. Results of a multinational study. AB - AIM: To compare the efficacy and tolerability of pantoprazole 40 mg and omeprazole MUPS 40 mg in patients with moderate to severe gastroesophageal reflux disease (GERD). METHODS: In this randomized, double-blind, parallel-group, multicenter study conducted in Austria, Germany, Portugal, Switzerland and The Netherlands, patients with endoscopically confirmed moderate to severe GERD (Savary/Miller esophagitis grade II/III) were enrolled. They received a once daily dose of either 40 mg pantoprazole or 40 mg omeprazole MUPS. Healing was determined by endoscopy after 4 weeks of treatment. If patients were not healed, treatment was extended for another 4 weeks. An additional endoscopy was performed in these cases after 8 weeks of treatment. Healing was determined by endoscopy after 4 and 8 weeks. In addition, treatment effect on symptoms was evaluated by the investigator using a questionnaire assessing heartburn, reflux regurgitation and pain on swallowing at each visit, as well as by a self-administered questionnaire comprising further 24 gastrointestinal symptoms. Analyses were performed for the intention-to-treat (ITT) and the per-protocol (PP) population. In addition, patients with high compliance (HC: 90% or = 5 metals was 26%, 36%, 15%, 12%, 6%, and 5%, respectively. Reactivity was most frequent to nickel (73%), followed by titanium (42%), cadmium (18%) gold (17%), palladium (13%), lead (11%), beryllium (9%), inorganic mercury (8%), tin (8%), and phenylmercury (6%). All patients (n=15) with confirmed or suspected nickel allergy were positive in MELISA, while patients with no suspicion of nickel allergy were either negative (n=6) or very low positive (n=4) in MELISA. MELISA reactivity is directly dependent on lymphocyte concentration: the higher the lymphocyte concentration per test, the stronger the reactivity. Concentrations of inorganic mercury > 0.5 microg/ml cause non antigen-specific (mitogenic) reactions in a majority of patients. The reproducibility rate was 94% using a cut-off of Stimulation Index > or = 3 or 99% using a cut-off of > or = 5. While the absolute intra- and interassay Stimulation Index values may vary, the qualitative results are highly reproducible. CONCLUSION: The MELISA Test is reproducible, sensitive, specific, and reliable for detecting metal sensitivity in metal-sensitive patients. PMID- 12743535 TI - Dental amalgam as one of the risk factors in autoimmune diseases. AB - BACKGROUND: Experimental and clinical data published recently show that dental amalgam can give rise to undesirable immunological responses in susceptible individuals. In genetically susceptible strains of experimental animals, mercury and silver can induce autoimmune responses. Sera of patients sensitive to mercury were found to have a higher incidence of autoantibodies relative to controls. OBJECTIVE: The aim of this study was to determine possible presence of antinuclear SSB/La autoantibodies after the in vitro stimulation of peripheral blood lymphocytes with HgCl2. METHODS: Lymphocytes were obtained from patients with autoimmune thyroiditis and increased response to mercury in vitro. Mononuclear cells were cultivated for 6 days with 100 microl HgCl2 solution or with pure medium and the levels of antinuclear autoantibodies SSB/La were assayed by a commercial SSB/La ELISA kit. RESULTS: Increased production of SSB/La autoantibodies in the media following stimulation of peripheral blood lymphocytes with HgCl2 was found in all cases. Using the Student's paired test, the results were significant on the p=0.05 significance level. CONCLUSION: Results imply that, in some patients with thyroiditis, mercury from dental amalgam can stimulate the production of antinuclear antibodies. Dental amalgam may be a risk factor in some patients with autoimmune disease. PMID- 12743536 TI - Effects of cold stress on morphine-induced nitric oxide production and mu-opiate receptor gene expression in Mytilus edulis pedal ganglia. AB - OBJECTIVES: Subjecting the marine bivalve Mytilus edulis to an immediate temperature change has been shown to rapidly alter the animals' ganglionic monoamine levels, as well as its ciliary activity. Recently, we extended this observation to include the organism's ganglionic mu opiate receptor and morphine levels. In the past, we demonstrated that M. edulis ganglionic mu receptors exposed to morphine was coupled to the immediate release nitric oxide (NO). In this study, we measured morphine-induced NO release in M. edulis subjected to acute cold stress. METHODS: NO release was monitored with an NO-selective microprobe. Temporal changes in mu opiate receptor expression were also examined over 24 hours. RESULTS: In this study, we demonstrate that after 12h cold exposure (4 degrees C from 24 degrees C), the estimated relative mu opiate receptor (MOR) gene expression in M. edulis pedal ganglia, measured by real-time PCR, did not differ significantly from the control group (1.23+/-0.25, p>0.05). However, the measured M. edulis pedal ganglia MOR expression demonstrated that ganglia significantly (0.77+/-0.05, p<0.001) down regulated their mu opiate receptor mRNA expression after 24h exposure to the cold water. The mean value for control animal (24 degrees C, n=14) morphine-stimulated NO release was 36.7 +/- 9.8 nM. Morphine additions to cold-treated tissues (4 degrees C, n=7) produced an average of 6.7 +/- 4.9 nM NO, which was a statistically significant difference between 25 degrees C and 4 degrees C animals (p=0.025). CONCLUSION: The study further demonstrates that mu opiate receptor expression is coupled to NO release. PMID- 12743537 TI - Real-time sonography for screening of gallbladder motility in diabetic patients: relation to autonomic and peripheral neuropathy. AB - OBJECTIVES: Diabetes mellitus is known as one of the factors causing the cholesterol gallstone. Gallstone incidence is about 30% in diabetic patients over 20 years of age. Pathophysiology is still not clear. The aim of the present study was to investigate gallbladder (GB) functions in diabetic patients and determine its relationship with peripheral and autonomic neuropathy. DESIGN: Study was performed between October 2001 and may 2002 in fifty-one diabetic patients of similar age and weight. Diabetic patients (n=51) were chosen randomly among diabetic patients, who were being followed in Diabetes Out-patient clinics of Selcuk University, Meram Medical Faculty. Twenty-eight control subjects were chosen from healthy volunteers. We measured fasting and post-prandial gallbladder volumes and ejection fractions by real-time ultrasonography. The patients were divided into three groups; group A (n=18) had no diabetic autonomic and peripheral neuropathy, group B (n=13) had diabetic peripheral neuropathy, group C (n=13) had diabetic autonomic neuropathy. RESULTS: No significant difference in any biochemical parameters between diabetic and control group could be found. Fasting gallbladder volume was significantly higher in the diabetic group (5.31 +/- 0.28 cm(3)) compared to control group (4.19 +/- 0.25 cm(3), p<0.01). But there was no difference within diabetic subgroups. Gallbladder ejection fraction was significantly reduced in diabetic patients in groups B and C (29.7 +/- 1.43%, 28.7 +/- 1.28%) compared to group A (44.8 +/- 2.4%; p<0.05, p<0.025 respectively). CONCLUSIONS: Cholesterol crystal formation as a result of increased gallbladder volume and decreased ejection fraction in diabetic patients may result from hypotonicity and stasis and thus this may lead to gallstones. PMID- 12743538 TI - Neuroendocrine alterations in lung cancer patients. AB - BACKGROUND: A number of qualitative and quantitative changes in hormonal secretion pattern have been found in subjects suffering from neoplastic disease. The aim of this study was to evaluate the presence of alterations in neuro endocrine system function and in the pattern of endocrine secretion in patients with lung cancer. METHODS: Cortisol, melatonin, growth hormone (GH), insulin-like growth factor I (IGF-I), thyrotropin-releasing hormone (TRH), thyroid-stimulating hormone (TSH), and free thyroxine (FT4) serum levels were measured on blood samples collected every four hours for 24 hours from ten healthy old subjects aged 65-79 years (mean age +/- s.e. 67.28 +/- 3.11) and from ten subjects suffering from untreated non small cell lung cancer aged 65-78 years (mean age +/ s.e. 68.57 +/- 1.81). Areas under the curve and mean diurnal and nocturnal levels were compared and the presence of circadian rhythmicity was evaluated. RESULTS: When hormone levels were expressed as area under the curve GH levels were higher (p=0.004) and IGF-I levels were lower (p=0.006) in patients with lung cancer than in normal subjects. The evaluation of melatonin/cortisol ratio in all subjects showed a significant difference between the control group and the group of cancer patients (p<0.05). When we compared mean diurnal levels (mean of 06.00 10.00-14.00h) GH levels were higher (p<0.0001) and IGF I levels were lower (p<0.0001) in cancer patients; when we compared mean nocturnal levels (mean of 18.00-22.00-02.00h) cortisol (p=0.03), TRH (p=0.02), and GH (p=0.001) levels were higher in cancer patients, while melatonin (p=0.04), TSH (p=0.04) and IGF I (p<0.0001) levels were higher in control subjects. A clear circadian rhythm was validated for time related changes of cortisol, melatonin, TRH, TSH and GH in control subjects and for time related changes of melatonin in cancer patients. CONCLUSION: These data suggest that lung cancer patients show alterations of hormone secretion and neuroendocrine system function. PMID- 12743539 TI - Reduction of cisplatin-induced anemia by the pineal indole 5-methoxytryptamine in metastatic lung cancer patients. AB - OBJECTIVE: It has been demonstrated that the hematopoiesis is under a neuroendocrine control, namely mediated by the pineal gland. The pineal indole melatonin (MLT) has appeared to exert thrombopoietic and lymphopoietic activity, whereas it has no relevant effect on red cell differentiation. The present study was performed to evaluate the influence of another pineal indole, the 5 methoxytryptamine (5-MTT) on red cell line and hemoglobin production. MATERIALS & METHODS: The study was carried out in metastatic lung cancer patients who underwent a chemotherapeutic combination containing cisplatin, which is known to induce anemia. The study included 20 patients treated with cisplatin plus etoposide, who were randomized to receive chemotherapy alone or chemotherapy plus 5-MTT (1 mg/day orally at noon every day). RESULTS: Hemoglobin mean blood concentrations significantly decreased in both groups of patients. However, the decrease in hemoglobin levels observed in patients treated with chemotherapy alone was significantly higher with respect to that observed in patients concomitantly treated with 5-MTT. Moreover, the percent of patients who had no progressive disease on treatment was significantly higher in the group treated with chemotherapy plus 5-MTT. CONCLUSIONS: Even though the low number of patients does not allow us to draw define conclusions, these preliminary results would show that the concomitant administration of 5-MTT may reduce cisplatin-induced anemia in cancer patients, by suggesting a hematopoietic activity of 5-MTT on red cell line differentiation and hemoglobin production. Moreover, the study would suggest that 5-MTT, as well as previously demonstrated for MLT, may also enhance the cytotoxic activity of cancer chemotherapy. PMID- 12743540 TI - Effects of melatonin on testicular tissue nitric oxide level and antioxidant enzyme activities in experimentally induced left varicocele. AB - OBJECTIVES: The pathophysiology of the testicular damage in varicocele have not been completely understood. There are several studies concerning the effects of increased seminal reactive oxygen species (ROS) and the role of nitric oxide (NO) in infertile patients with varicocele and antioxidants have been used successfully to decrease oxidative stress in testis. In this study, we determined the effects of melatonin on testicular NO and antioxidant enzyme levels and investigated whether melatonin can prevent or diminish testicular damage in adult rats with experimentally induced unilateral varicocele. METHODS: Thirty adult male Wistar rats were randomly divided into three groups. In group I, using midline incision left renal vein was exposed but not tied. In group II, left renal vein was partially ligated to create varicocele. In group III, after creation of varicocele, daily and fresh-prepared melatonin was administered intraperitoneally for 4 weeks. Histopathological examination was performed and tissue malondialdehyde (MDA), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and tissue NO levels were determined. RESULTS: In group II, there were severe degenerative changes of the germinal epithelium and atrophy of the seminiferous tubules in left testis. In group III, in which rats received melatonin intraperitoneally, there were focal areas showing decrease in spermatogenesis and some degenerative changes. Similarly, the levels of ROS and NO were significantly increased in testicular tissue of rats in group II, whereas group III rat testes in which melatonin was administered, showed increased antioxidant enzyme activity and decreased NO levels. CONCLUSION: Infertility related to varicocele has multiple pathophysiological mechanisms and the effect of ROS, NO and antioxidant defense system are contributing factors in the disease state. Melatonin, a potent free radical scavenger, administration might be helpful in the prevention or at least, in the delay of the severe effects of varicocele on testicular tissue. PMID- 12743541 TI - Steroid modulation of angiotensin II action in the rat anterior pituitary gland. AB - OBJECTIVES: The present study was designed to test whether various steroid hormones modulate differently angiotensin II (AngII) action in the anterior pituitary in males and females. MATERIAL AND METHODS: Adult female and male rats were treated with one of the following substances: oil (control), pregnenolone sulfate (PREG-S), 17beta-estradiol benzoate (E2,) progesterone (P), or dehydroepiandrosterone sulfate (DHEA-S), given in intraperitoneal injections for five days in dose of 50 microg per animal per day. Because AngII is known to act in the anterior pituitary through the phosphatidiloinositol breakdown, thus increasing the level of inositol-1,4,5-trisphosphate (IP3), the IP3 concentration was determined 24 hours after the injection in the anterior pituitary homogenate exposed to AngII. RESULTS: In control animals (without steroids) AngII stimulated concentration of IP(3) stronger in females than in males. E(2) and DHEA-S enhanced AngII effects in both males and females. PREG-S increased AngII-induced IP(3) concentration in females, but not males. Progesterone raised AngII effect on IP(3) concentration in males, only when high concentrations of peptide were used. CONCLUSION: These results indicate that pituitary sensitivity to AngII stimulation is modulated by steroid hormones and is related to the gender of the animal. PMID- 12743542 TI - Responses of the HPA axis after chronic variable stress: effects of novel and familiar stressors. AB - OBJECTIVES: We examined the role that novelty plays in determining interactions between chronic and acute stress, when both the chronic and acute stressors emphasize emotional processing (i.e. stressful stimuli that do not present immediate threats to somatic homeostasis, and are processed primarily by limbic and forebrain circuits). METHODS: Rats were exposed to a chronic variable stress (CVS) regimen, and were subsequently tested to evaluate responses to novel and familiar acute stressors. One group was exposed to CVS that included restraint, and was then tested with this familiar stressor. Another group was exposed to CVS that did not include restraint, and were tested with restraint as a novel stressor. Additional rats were not chronically stressed. Plasma adrenocorticotrophic hormone (ACTH) and corticosterone (CORT) were assayed. RESULTS: When the rats were exposed to familiar acute stress after CVS, ACTH responses were blunted. The ACTH responses were normal in the rats that were tested with novel acute stress--the responses resembled those of rats that had no prior stress experience. CORT responses did not differ between the groups, regardless of stress history. CONCLUSIONS: Despite the fact that all the chronic and acute stressors emphasized emotional processing of aversive stimuli, and thus likely involved overlapping limbic and forebrain circuits, the hormonal responses differed depending upon familiarity with the acute stressor. Further research is required to identify the neuronal mechanisms that mediate these differing responses to novel and familiar emotional stressors. PMID- 12743547 TI - First do no harm: managing antihistamine impairment in patients with allergic rhinitis. AB - Antihistamines are effective medications that have been used for decades in the management of allergic rhinitis; however, they may be administered or selected in an inappropriate fashion and may be the source of drug-related morbidity. Our objective is to present relevant background information and an expert consensus statement on the use of antihistamines in treatment of allergic rhinitis. In July 2002, 14 experts in allergy, clinical immunology, pharmacology, and impairment assessment were invited to participate in a roundtable conference to present current concepts and develop a consensus statement on the clinical management of allergic rhinitis with antihistamines. Many of the antihistamines used to treat allergic rhinitis, as well as the disease itself, may produce sedation, impairment, and reduced quality of life. Allergic rhinitis is more appropriately managed with the relatively nonimpairing second-generation antihistamines (eg, loratadine, desloratadine, cetirizine, and fexofenadine), because older agents (eg, diphenhydramine, chlorpheniramine, and brompheniramine) produce sedation and impairment and worsen sleep architecture. Although there is some debate surrounding the varying degrees of efficacy of second-generation antihistamines, it is known that some agents may produce varying levels of drowsiness or impairment, especially at higher than recommended doses. The differences with regard to safety among the second-generation antihistamines are smaller than are the differences between the first and second generations. A nonsedating, nonimpairing (even at higher than recommended doses), second-generation antihistamine is preferred for all patients, particularly those with a higher risk for the development of adverse effects. We recommend that primary care and specialist physicians, nurse practitioners, physician assistants, pharmacists, and all other health professionals involved in the diagnosis and treatment of allergic rhinitis follow this consensus document and share this information with patients for whom antihistamine therapy is recommended. In addition, further epidemiologic studies on the effects of antihistamines should be performed. PMID- 12743549 TI - Aspirin-induced asthma: advances in pathogenesis, diagnosis, and management. AB - In some asthmatic individuals, aspirin and other nonsteroidal anti-inflammatory drugs (NSAIDs) that inhibit cyclooxygen-ase 1 (COX-1) exacerbate the condition. This distinct clinical syndrome, called aspirin-induced asthma (AIA), is characterized by an eosinophilic rhinosinusitis, nasal polyposis, aspirin sensitivity, and asthma. There is no in vitro test for the disorder, and diagnosis can be established only by provocation challenges with aspirin or NSAIDs. Recent major advances in the molecular biology of eicosanoids, exemplified by the cloning of 2 cysteinyl leukotriene receptors and the discovery of a whole family of cyclooxygenase enzymes, offer new insights into mechanisms operating in AIA. The disease runs a protracted course even if COX-1 inhibitors are avoided, and the course is often severe, many patients requiring systemic corticosteroids to control their sinusitis and asthma. Aspirin and NSAIDs should be avoided, but highly specific COX-2 inhibitors, known as coxibs, are well tolerated and can be safely used. Aspirin desensitization, followed by daily aspirin treatment, is a valuable therapeutic option in most patients with AIA, particularly those with recurrent nasal polyposis or overdependence on systemic corticosteroids. PMID- 12743551 TI - Understanding exocytosis in immune and inflammatory cells: the molecular basis of mediator secretion. AB - Inflammatory cells secrete proteins from intracellular vesicles or granules by a process referred to either as exocytosis or as degranulation, which is common to all cell types. Exocytosis is a precise term that describes the process of granule or vesicular fusion with the plasma membrane and is accompanied by release of granule/vesicle contents to the cell exterior. This process is of particular significance with respect to tissue damage and remodeling in inflammatory diseases, inasmuch as these changes are the consequences of inflammatory cell activation and mediator elaboration. Despite its unifying importance to all inflammatory cell types, little is known about the precise molecular and intracellular mechanisms that regulate mobilization of secretory granules/vesicles and, ultimately, secretion of mediators from immune and inflammatory cells. This article reviews the mechanisms and molecules currently implicated at distal stages of exocytosis from eosinophils, neutrophils, mast cells, platelets, and macrophages. Conserved molecules identified among inflammatory cell types indicate a convergence of pathways leading to mediator secretion. The identification of essential molecules in the cascade of events leading to exocytosis is critical in the search for novel therapeutic targets aimed at modulating mediator secretion from these cell types. PMID- 12743553 TI - Antifungal therapy for allergic bronchopulmonary aspergillosis. PMID- 12743554 TI - In the eye of the patient. PMID- 12743555 TI - Early exposure to allergen: is this the cat's meow, or are we barking up the wrong tree? AB - Several recent studies have suggested that exposure to cat and dog allergen in infancy is protective against the subsequent risk of allergic sensitization and asthma. The methodologic problems to be overcome in clinical research addressing these hypotheses are complex. Appreciation of these studies requires an assessment of the design and adequacy of variables measuring exposures, outcomes, and confounders. It includes understanding the role of effect modification. This article discusses some of the epidemiologic issues in interpreting these studies. Review of relevant epidemiology demonstrates that much research remains to be performed before these interesting hypotheses are proved or disproved. PMID- 12743556 TI - Schistosoma mansoni infection is associated with a reduced course of asthma. AB - BACKGROUND: Helminthic infections decrease skin reactivity to indoor allergens, but data on whether they influence asthma severity are lacking. OBJECTIVE: This study evaluated the course of asthma in patients with and without Schistosoma mansoni infection. METHODS: Asthmatic subjects were enrolled from 3 low socioeconomic areas: a rural area endemic for schistosomiasis (group 1) in addition to a rural area (group 2) and a slum area (group 3), both of which were not endemic for schistosomiasis. A questionnaire on the basis of the International Study of Asthma and Allergies in Childhood study was applied in these 3 areas, and from each area, 21 age- and sex-matched asthmatic subjects were selected for a prospective 1-year study. Pulmonary function tests, skin prick tests with indoor allergens, stool examinations, and serum evaluations were performed in these subjects. Every 3 months, the subjects were evaluated for asthma exacerbation through physical examination, and a questionnaire regarding asthma symptoms and use of antiasthma medicine was administered. RESULTS: The prevalence of S mansoni infection was greater in group 1 compared with in groups 2 and 3 (P <.0001), whereas the frequency of other helminth and protozoa infections was similar among the 3 groups. The frequency of positive skin test responses to indoor allergens was less (19.0%) in group 1 subjects relative to those in group 2 (76.2%) and group 3 (57.1%; P <.001). The frequencies of symptoms, use of antiasthma drugs, and pulmonary abnormal findings at physical examination were less in group 1 subjects than in group 2 and 3 subjects (P =.0001). CONCLUSION: Our results suggest that S mansoni infection is associated with a milder course of asthma. PMID- 12743557 TI - Anti-inflammatory effect of itraconazole in stable allergic bronchopulmonary aspergillosis: a randomized controlled trial. AB - BACKGROUND: Allergic bronchopulmonary aspergillosis (ABPA) complicates chronic asthma and results from hypersensitivity to the fungus Aspergillus fumigatu s, causing an intense systemic immune response and progressive lung damage. OBJECTIVE: We sought to determine whether treatment with the antifungal agent itraconazole reduced eosinophilic airway inflammation in subjects with ABPA. METHODS: A randomized, double-blind, placebo-controlled trial was performed in stable subjects with ABPA (n = 29). Subjects received 400 mg of itraconazole per day (n = 15) or placebo (n = 14) for 16 weeks. All subjects were reviewed monthly with history, spirometry, and sputum induction to measure airway inflammation, serum total IgE and IgG levels to A fumigatu s, and blood eosinophil counts. RESULTS: By using regression analysis in a random-effects model, subjects receiving itraconazole had a decrease in sputum eosinophils of 35% per week, with no decrease seen in the placebo arm (P <.01). Sputum eosinophil cationic protein levels decreased with itraconazole treatment by 42% per week compared with 23% in the placebo group (P <.01). Itraconazole reduced systemic immune activation, leading to a decrease in serum IgE levels (310 IU/mL) compared with levels seen in the placebo group (increase of 18 IU/mL, P <.01) and a decrease in IgG levels to A fumigatu s (15.4 IU/mL) compared with levels seen in the placebo group (increase of 3.7 IU/mL, P =.03). There were fewer exacerbations requiring oral cortico-steroids in those treated with itraconazole compared with in the placebo group (P =.03). CONCLUSION: Itraconazole treatment of subjects with stable ABPA reduces eosinophilic airway inflammation, systemic immune activation, and exacerbations. These results imply that itraconazole is a potential adjunctive treatment for ABPA. PMID- 12743558 TI - Effects of suplatast tosilate on allergic eosinophilic airway inflammation in patients with mild asthma. AB - BACKGROUND: Bronchial asthma is a chronic inflammatory disease characterized mainly by infiltration of the airway mucosa by various inflammatory cells, notably eosinophils. T(H)2-type cytokines are suggested to be deeply involved in the pathogenesis of asthma. OBJECTIVE: We sought to determine the suppressive effects of suplatast tosilate, an inhibitor of T(H)2-type cytokines, on eosinophilic inflammation of the bronchial mucosa in patients with mild asthma. METHODS: Airway hyperresponsiveness tests, pulmonary function tests, eosinophil measurements in induced sputum, and bronchial mucosa biopsies were performed before and after treatment with suplatast tosilate for 6 weeks in 15 patients with mild asthma and in 13 control patients with mild asthma not receiving suplatast tosilate. This study was performed as a case-controlled open study. RESULTS: In the treatment group a significant improvement in the provocation concentration of histamine was observed (P <.05). Improvements in peak expiratory flow (P <.01) and in symptom score (P <.05) were also noted in the suplatast tosilate-treated group. Moreover, the average number of infiltrating eosinophils and EG2(+) cells significantly decreased (both P <.05), as did the ratios of eosinophils and EG2(+) cells in sputum (both P <.01). The average number of CD4(+) and CD25(+) T lymphocytes also decreased (both P <.05). CONCLUSION: Suplatast tosilate appears to inhibit allergic airway inflammation mediated by T(H)2-type cytokine and to improve clinical symptoms in patients with mild asthma. PMID- 12743559 TI - Qualitative research-enhanced understanding of patients' beliefs: results of focus groups with low-income, urban, African American adults with asthma. AB - BACKGROUND: Qualitative research is an important research methodology for understanding the health beliefs and attitudes of patients. These beliefs and attitudes have been proposed as partial explanations for low adherence to medical therapy and the consequent high burden of morbidity from asthma among low-income urban minorities. OBJECTIVE: The purpose of this study was to explore barriers to adherence to inhaled corticosteroids (ICSs), health beliefs regarding asthma and its treatment, and opinions about providers and clinical research among low income groups. METHODS: Three focus groups were conducted with 15 low-income, urban, African American adults with persistent asthma. These focus group sessions were audiotaped, transcribed verbatim, and coded by using qualitative analytic techniques. RESULTS: Health beliefs that influenced adherence included patients' reliance on their assessment of asthma control over that of the health provider and concern over the adverse effects of ICS therapy. Adherence was also adversely affected by social obligations and insurers' approval policies and restricted formularies. Although mistrust of the medical establishment was evident, members generally expressed a willingness to participate in nonpharmacologic clinical research and the belief that research was beneficial. However, they had strong preferences for where the research should be conducted. CONCLUSIONS: Although barriers exist to ICS adherence in patient populations, many of these can be addressed during patient-clinician interactions. Qualitative research is an important tool for formulating hypotheses for improving ICS adherence that can then be tested in the future by using quantitative research methods. PMID- 12743561 TI - Smoking might be a risk factor for contact allergy. AB - BACKGROUND: Contact allergy is a major public health problem in industrialized countries. Hitherto, known risk factors for contact allergy have mainly included increased exposure to allergens. There are no published data on the relation between smoking and contact allergy. OBJECTIVE: To investigate the association between smoking and contact allergy. METHODS: The study population comprised a cross-sectional, general population-based sample of 15- to 69-year-old persons living in Copenhagen, Denmark. A total of 1056 persons (73.6% of the invited) were given a patch test (TRUE test). Contact allergy was defined as a positive patch test result to at least 1 of 23 allergens. Nickel contact allergy was defined as a positive patch test reaction to nickel. Allergic nickel contact dermatitis was defined as a history of eczema on exposure to metallic objects and a positive patch test reaction to nickel. A detailed smoking history was obtained in a questionnaire. RESULTS: Contact allergy (adjusted odds ratio, 1.8; 95% CI, 1.2 to 2.9), nickel contact allergy (adjusted odds ratio, 2.7; 95% CI, 1.4 to 5.2), and allergic nickel contact dermatitis (adjusted odds ratio, 3.0; 95% CI, 1.5 to 6.2) were significantly associated with a smoking history of more than 15 pack-years. Moreover, these associations showed a significant dose-response relation, and they were independent of sex, age, and exposure to nickel, as reflected by a history of ear piercing. CONCLUSIONS: These data raise the hypothesis that smoking increases the risk of contact allergy. Further epidemiologic studies and investigations into the possible mechanisms are warranted. PMID- 12743560 TI - Identification of cross-reactive and genuine Parietaria judaica pollen allergens. AB - BACKGROUND: The weed Parietaria judaica is one of the most important pollen allergen sources in the Mediterranean area. OBJECTIVE: We sought to identify P judaica pollen allergen, which might be used to serologically distinguish genuine Parietaria sensitization and cross-reactivity to allergens from other weed species (eg, mugwort and ragweed). METHODS: The allergen profile of P judaica IgE reactive sera from weed pollen-sensitized allergic individuals from the Mediterranean region (n = 36) with high Parietaria pollen exposure and from weed pollen-allergic patients with little or no Parietaria exposure (Austria, n = 42; Scandinavia, n = 8; United States, n = 19) was established by CAP FEIA measurements and by IgE immunoblot inhibition experiments with recombinant allergens. RESULTS: The majority (83%) of the Mediterranean weed pollen-allergic patients mounted high IgE antibody levels (mean specific IgE, 20.89 kUA/L) against recombinant (r) Par j 2, whereas only 7% of the non-Mediterranean weed allergic patients showed low IgE reactivity to rPar j 2 (mean specific IgE, 1.03 kUA/L). The cytoskeletal protein profilin and a 2-EF-hand calcium-binding allergen were identified as cross-reactive Parietaria allergens, which were recognized preferentially by Parietaria -positive, non-Mediterranean weed pollen allergic patients. CONCLUSION: rPar j 2 might be used as a diagnostic marker allergen to identify weed pollen-allergic patients who are genuinely sensitized against Parietaria pollen and thus would be particularly suited for specific immunotherapy with Parietaria pollen extract. PMID- 12743562 TI - Specific immunotherapy with a standardized latex extract in allergic workers: a double-blind, placebo-controlled study. AB - BACKGROUND: Preventive measures have been proposed to reduce the risk of sensitization to natural rubber latex (NRL), but this is not always feasible. OBJECTIVE: The aim of this study was to determine the efficacy and safety of specific immunotherapy with a standardized latex extract in sensitized workers. METHODS: Twenty-four patients allergic to NRL with contact urticaria (n = 8) and rhinitis or asthma (n = 16) were included (16 in the active group and 8 in the placebo group). Treatment started in a cluster immunotherapy protocol, with injections every week for 3 months and then every other week for another 3 months. RESULTS: Patients in the active group had significantly lower values than patients in the placebo group in skin terms of reactivity to NRL (P <.01), rubbing test results (P =.047), and latex glove use test results (P =.046) after 6 months of treatment. There were no significant differences between the active and placebo groups in symptom scores, use of medication, self-assessment, or methacholine test results either before or after treatment. Differences in nasal and bronchial symptoms during specific inhalation challenges (P = not significant and P =.05, respectively) were observed in favor of the active group. In the active group 32 systemic reactions were observed (8% of doses), mostly during the build-up period, being more frequent in patients with respiratory symptoms (P =.004). All reactions responded promptly to treatment. CONCLUSION: Clinical efficacy was shown mainly on cutaneous symptoms, although an improvement in rhinitis and asthma symptoms was also observed during specific inhalation challenges. Latex-specific immunotherapy might be a useful approach for the treatment of latex allergy in sensitized workers. PMID- 12743563 TI - Reduced risk of atopy among school-age children infected with geohelminth parasites in a rural area of the tropics. AB - BACKGROUND: Childhood infections might protect against the expression of atopy. Geohelminths are among the most prevalent infections of childhood and might contribute to the low prevalence of allergic disease reported from rural areas of the tropics. OBJECTIVE: We sought to establish whether geohelminth infections protect against atopy and to explore whether this protection is dependent on infection chronicity. METHODS: The risk of atopy (measured by means of allergen skin test reactivity) associated with active geohelminth infections (measured by means of the presence of eggs in stool samples) or with chronic geohelminth infections (measured by means of high levels [>/=3564 IU/mL] of total serum IgE or the presence of detectable anti-Ascaris lumbricoides IgG4 antibodies) was investigated in an analytic cross-sectional study conducted among school-age children attending rural schools in Pichincha Province in Ecuador. RESULTS: A total of 2865 children aged 5 to 19 years from 55 schools was examined. Active infection with any geohelminth and infections with A lumbricoides or Ancylostoma duodenale were associated with significant protective effects against allergen skin test reactivity. Children with the highest levels of total IgE or with anti A lumbricoides IgG4 antibodies were protected against skin test reactivity also, and the protective effects of high IgE or anti-A lumbricoides IgG4 and or active geohelminth infections were statistically independent. CONCLUSION: Active infections with geohelminth parasites and the presence of serologic markers of chronic infections (high levels of total serum IgE or anti-A lumbricoides IgG4) are independent protective factors against allergen skin test reactivity among school-age children living in an endemic region of the rural tropics. PMID- 12743564 TI - Association of polymorphisms in the collagen region of SP-A2 with increased levels of total IgE antibodies and eosinophilia in patients with allergic bronchopulmonary aspergillosis. AB - BACKGROUND: Studies from our group have shown a protective role of pulmonary surfactant protein A (SP-A) against lung allergy and infections caused by Aspergillus fumigatus. OBJECTIVE: Present study investigated the association of polymorphisms in the collagen region of SP-A1 and SP-A2 (genes encoding SP-A) with allergic bronchopulmonary aspergillosis (ABPA) and its clinical markers. METHODS: Genomic DNA was extracted from blood samples of patients with ABPA and age-matched, unrelated control subjects. The polymorphisms were detected by means of PCR amplification and sequencing of the collagen region of SP-A1 and SP-A2. RESULTS: Two exonic (SP-A2 G1649C and SP-A2 A1660G, 10 patients and 11 control subjects) and 2 intronic (SP-A2 T1492C, 8 patients and 8 control subjects; SP-A1 C1416T, 5 patients and 7 control subjects) polymorphisms in the collagen region of SP-A2 and SP-A1 showed significant association with patients with ABPA. A significantly higher frequency of the AGA allele (A1660G) of SP-A2 was observed in patients with ABPA in comparison with control subjects (P =.0156, odds ratio [OR] = 4.78, 95% CI = 1.23 < OR < 18.52). This polymorphism, when existing along with a nonredundant polymorphism, SP-A2 G1649C (Ala91Pro) resulted in a stronger association with ABPA (A1660G and G1649C: P =.0079, OR = 10.4, 95% CI = 1.62 < OR < 66.90). Patients with ABPA with GCT and AGG alleles showed significantly high levels of total IgE and percentage eosinophilia versus patients with ABPA with CCT and AGA alleles. CONCLUSION: The results indicated that SP-A2 G1649C and SP A2 A1660G, polymorphisms in the collagen region of SP-A2, might be one of the contributing factors to genetic predisposition and severity of clinical markers of ABPA. PMID- 12743565 TI - Experimental rhinovirus challenges in adults with mild asthma: response to infection in relation to IgE. AB - BACKGROUND: Although most children and young adults with asthma are atopic, exacerbations of asthma are frequently associated with viral respiratory tract infections, especially those caused by rhinovirus (HRV). OBJECTIVE: Young atopic adults with mild asthma were evaluated before and during an experimental HRV infection to test the hypothesis that airway inflammation before virus inoculation may be a risk factor for an adverse response to HRV. METHODS: Experimental HRV infections were evaluated in 16 allergic volunteers with mild asthma and 9 nonatopic control patients (age, 18 to 30 years). Before virus inoculation, each participant was screened with tests for lung function, prick skin tests for sensitization to common aeroallergens, measurements of total serum IgE, and serum neutralizing antibody to rhinovirus-16 (the serotype used for inoculation). The response to infection was monitored for 21 days by using symptom diary cards, tests for lung function, and markers of airway inflammation in nasal washes, blood, and expired air. RESULTS: During the infection, asthmatic patients had cumulative upper and lower respiratory tract symptom scores that were significantly greater over the course of 21 days than scores from the control patients. At baseline, the asthmatic patients also had increased sensitivity to methacholine and significantly lower values for FEV(1) (percent predicted) than the control patients (geometric mean and intraquartile values: 87% [79% to 91%] for the asthmatic patients and 101% [90% to 104%] for the control patients, P <.03). Among the patients with mild asthma, 6 had levels of total serum IgE that were substantially elevated (range, 371 to 820 IU/mL) compared with 10 who had lower levels (range, 29 to 124 IU/mL). Those with high levels of IgE had significantly greater lower respiratory tract symptom scores during the initial 4 days of the infection than the low IgE group. They also had higher total blood eosinophil counts at baseline, increased eosinophil cationic protein in their nasal washes (>200 ng/mL), and augmented levels of expired nitric oxide at baseline and during peak cold symptoms. In contrast, levels of soluble intracellular adhesion molecule-1 in nasal wash supernatants from the asthmatic patients with high IgE were diminished, both at baseline and during the infection. CONCLUSIONS: The reduced lung function and increased markers of inflammation observed before virus inoculation in the asthmatic patients who had high levels of total serum IgE may be risk factors for an adverse response to infections with HRV. PMID- 12743566 TI - Abortive pollen germination: a mechanism of allergen release in birch, alder, and hazel revealed by immunogold electron microscopy. AB - BACKGROUND: Pollen from early-flowering trees (eg, birch, alder, hazel) represent major seasonal allergen sources. The effects of rain on the release of allergens from tree pollen has thus far not been studied at the ultrastructural level. OBJECTIVE: This study was designed to investigate the effects of rain on the morphology of pollens from early-flowering trees and of potential rain-induced mechanisms of allergen release. METHODS: Freshly collected pollen grains (birch, alder, and hazel) were exposed under controlled conditions to rainwater. Changes of pollen morphology and the release of allergens were analyzed by scanning electron microscopy. The release of allergen-bearing submicronic particles was studied by field emission scanning electron microscopy and transmission electron microscopy in conjunction with immunogold staining by using antibodies with specificity for the major allergens. RESULTS: Scanning electron microscopy showed that freshly isolated pollen grains from birch, alder, and hazel have abortive germination in rainwater. Abortive pollen germination is characterized by the formation of short pollen tubes, which rupture at their tips and release micronic and submicronic particles containing major allergens. Immunogold transmission electron microscopy provided evidence that the allergens are transported through the pollen tubes during germination. CONCLUSIONS: Rainwater-induced release of allergen-bearing submicronic particles from abortively germinated tree pollens may represent a mechanism of allergen release, with important implications on the induction of asthma as well as on current methods for measuring environmental allergen exposure. PMID- 12743567 TI - Ligation of intercellular adhesion molecule 3 inhibits GM-CSF production by human eosinophils. AB - BACKGROUND: Intercellular adhesion molecule 3 (ICAM-3) has recently been identified on the surface of eosinophils. OBJECTIVE: The purpose of this study was to characterize ICAM-3 expression on eosinophils in response to cytokines and to determine whether ligand binding of ICAM-3 modulates inflammatory responses of eosinophils, as it does in other leukocytes. METHODS: To determine effects of ICAM-3 on eosinophil function, we isolated human eosinophils and used a monoclonal antibody directed against the epitope of ICAM-3 that binds to leukocyte-function antigen-1 to mimic binding of ICAM-3 and this natural ligand. We measured granulocyte-macrophage colony stimulating factor (GM-CSF) production by unstimulated eosinophils and eosinophils stimulated with ionomycin (1 micromol/L), both in the presence and absence of this anti-ICAM-3 antibody. RESULTS: We found that 99% of eosinophils expressed ICAM-3, regardless of whether allergic symptoms were present or absent. Expression of ICAM-3 was not enhanced by proinflammatory cytokines. Expression of ICAM-3 was reduced in apoptotic cells and in cells incubated with the combination of GM-CSF and tumor necrosis factor alpha (n = 3). Antibody binding of ICAM-3, which mimics leukocyte-function antigen-1 binding, had no effect on baseline GM-CSF production but reduced by 80% the production of GM-CSF stimulated by ionomycin (control 1969 pg/mL +/- 1259 SD versus anti-ICAM-3 396 pg/mL +/- 207 SD, n = 8) and reduced GM-CSF mRNA content. CONCLUSIONS: ICAM-3 is highly expressed on the surface of human eosinophils, and downregulation of GM-CSF production by anti-ICAM-3 mAb suggests that ICAM-3 ligation may inhibit eosinophil inflammatory responses and survival. PMID- 12743568 TI - CysLT1 receptor upregulation by TGF-beta and IL-13 is associated with bronchial smooth muscle cell proliferation in response to LTD4. AB - BACKGROUND: Airway remodeling is a feature of chronic asthma. It involves a number of structural changes, including bronchial smooth muscle cell (BSMC) hyperplasia and hypertrophy. Cysteinyl leukotrienes (cysLTs) have been suggested to play a role in airway remodeling in addition to their numerous other physiopathologic effects. OBJECTIVES: This work was aimed at characterizing the potential modulation of CysLT1 receptor expression by cytokines and the eventual functional relevance of this modulation. METHODS: Expression of CysLT1 receptor was measured by flow cytometry and immunofluorescence microscopy. Transcripts were measured by RT-PCR and BSMC proliferation by crystal violet staining. RESULTS: When human BSMC were exposed to transforming growth factor (TGF)-beta, IL-13, or IFN-gamma, their expression of CysLT1 receptor was significantly augmented in a time- and concentration-dependent manner. Interestingly, IL-4 had no significant effect on CysLT1 receptor expression in BSMC. Moreover, IL-13 and IFN-gamma but not TGF-beta were able to increase CysLT1 mRNA levels. Finally, when BSMC were pretreated with TGF-beta or IL-13 but not IFN-gamma, their responsiveness to LTD(4) was markedly enhanced in terms of BSMC proliferation. Whereas TGF-beta, IL-13, or LTD(4) alone had little effect on BSMC proliferation, preexposure of the cells to TGF-beta or IL-13 for 24 hours resulted in a significant increase in proliferation in response to LTD(4). The enhanced proliferation was totally prevented by pretreating the cytokine-primed BSMC with the selective CysLT1 receptor antagonist Montelukast. CONCLUSIONS: Taken together, our findings indicate a synergy between certain cytokines and cysLTs, mediated by the augmented expression of the CysLT1 receptor and subsequent LTD(4) triggered BSMC proliferation. These findings support a role for cysLTs in the airway remodeling observed in asthmatic patients and may provide a rationale for preventive and therapeutic intervention. PMID- 12743569 TI - Deficient prostaglandin E2 production by bronchial fibroblasts of asthmatic patients, with special reference to aspirin-induced asthma. AB - BACKGROUND: Regulation of prostaglandin synthesis and the activation of human airway fibroblasts associated with the remodeling of the bronchi play an important role in asthma. OBJECTIVE: We sought to assess the cyclooxygenase pathways in airway fibroblasts of patients with bronchial asthma. METHODS: Generation of prostaglandin E(2) (PGE(2)) and pros-taglandin D(2) (PGD(2)) by bronchial fibroblasts was measured by means of mass spectrometry in culture supernatants, and cyclooxgenases expression was estimated by means of RT-PCR and immunoblotting. The cells were isolated from 3 groups of subjects: nonasthmatic patients (n = 10), patients with aspirin-tolerant asthma (ATA, n = 9), and patients with aspirin-intolerant asthma (AIA, n = 7). RESULTS: The cytomix (LPS, 5 square g/mL; IL-1 square, 5 ng/mL; and TNF- square, 10 ng/mL; 18 hours) stimulated the production of prostaglandins. Asthmatic patients were characterized by low capacity to produce PGE(2) after cytomix stimulation. In the nonasthmatic patient group the mean PGE(2) production was 32 +/- 33 ng/mL (35 fold of the basic production), in the ATA group it was 16 +/- 18 ng/mL (16-fold), and in the AIA group it was only 5.3 +/- 3.6 ng/mL (4-fold). The mean concentration of PGD(2) for nonasthmatic patients, patients with ATA, and patients with AIA was 0.18 +/- 0.16 ng/mL (4.7-fold of the basic production), 0.18 +/- 0.14 ng/mL (4.2-fold), and 0.235 +/- 0.19 ng/mL (1.9-fold), respectively. The observed difference was not due to insufficient cyclooxygenase 2 expression because all groups had similar levels of its mRNA and protein. The patients with AIA had low expression levels of cyclooxygenase 1 protein but not of its mRNA. The PGE(2)/PGD(2) concentration ratio increased after cytomix stimulation in all groups but was significantly less in patients with AIA than in patients with ATA. CONCLUSIONS: Our results point to a deficient PGE(2) production under proinflammatory conditions in asthmatic airways. This could weaken local defensive mechanisms and promote cysteinyl leukotriene overproduction. PMID- 12743570 TI - Inhibition of allergic airways inflammation and airway hyperresponsiveness in mice by dexamethasone: role of eosinophils, IL-5, eotaxin, and IL-13. AB - BACKGROUND: Glucocorticoids inhibit allergen-induced airway eosinophilia and airway hyperresponsiveness (AHR). Whether glucocorticoids mediate their effects on AHR by inhibiting eotaxin and IL-5, 2 of the principal mediators of eosinophilia, or through IL-13, an important mediator of AHR, has not been established. OBJECTIVE: We sought to investigate the effects of glucocorticoids on airway eosinophilia and the expression of IL-5, eotaxin, and IL-13 in relation to the induction of AHR in a murine model of allergic asthma. METHODS: Dexamethasone (4 mg/kg) and mAbs against eotaxin (80 micro g/kg) and IL-5 (100 micro g/kg) singly and in combination were administered to immunized mice before antigen challenge. Airway responsiveness to methacholine was measured in anesthetized and mechanically ventilated animals. Eotaxin, IL-5, and IL-13 in bronchoalveolar lavage fluid (BALF), lung homogenates, or both were measured by means of ELISA. RESULTS: A single antigen challenge induced AHR that lasted at least 10 days. Eotaxin protein and mRNA levels increased in lung tissue but not in BALF after challenge. IL-5 protein and mRNA levels increased both in BALF and in lung tissue. Dexamethasone reduced airway eosinophilia, AHR, and protein and mRNA for eotaxin and IL-5. Anti-murine eotaxin and anti-IL-5 antibodies alone and in combination reduced the ovalbumin-induced airway eosinophilia significantly but failed to inhibit AHR. Both dexa-methasone and anti-IL-5/anti-eotaxin inhibited the increases in lung IL-13 levels after ovalbumin challenge to a similar extent. CONCLUSION: These findings suggest that the inhibition of AHR by the glucocorticoid dexamethasone does not appear to be explained by effects on eosinophilia, eotaxin, IL-5, or IL-13. PMID- 12743571 TI - Role of activator protein 1, nuclear factor-kappaB, and nuclear factor of activated T cells in IgE receptor-mediated cytokine expression in mature human mast cells. AB - BACKGROUND: On activation by cross-linking the high-affinity IgE receptor (FcepsilonRI), expression of TNF-alpha, IL-3, IL-5, and IL-13 is induced in human intestinal mast cells. OBJECTIVE: We sought to examine, for the first time, FcepsilonRI signaling in mature human mast cells. METHODS: Mast cells were isolated from intestinal tissue and cultured in the presence of stem cell factor. The cells were treated with specific inhibitors before stimulation by means of FcepsilonRI cross-linking. Cytokine mRNA expression was analyzed by means of RT PCR, and activation of signaling molecules was determined by means of immunocytochemistry, RT-PCR, Western blotting, and protein kinase C (PKC) assay. RESULTS: We found that nuclear factor-kappaB (NF-kappaB), as well as c-Fos and c Jun, the components of activator protein 1 (AP-1), are activated after FcepsilonRI cross-linking in human intestinal mast cells. Treatment of the cells with specific inhibitors revealed an involvement of NF-kappaB and nuclear factor of activated T cells, as well as the necessity of extracellular signal-regulated kinase 1/2 (ERK-1/2), PKC, and AP-1 for the induced cytokine gene expression. Consistently, we found that activation of c-Fos corresponds with the induced cytokine gene expression and that ERK-1/2, an activator of c-Fos, was activated in response to FcepsilonRI cross-linking. CONCLUSION: Our data on human mast cells show that the activity of ERK-1/2, PKC, and subsequent activation of AP-1 are necessary for the FcepsilonRI-mediated cytokine expression. Nuclear factor of activated T cells and NF-kappaB seem to be necessary for the induction of TNF alpha, IL-3, and IL-13 but are less important for the transcription of IL-5. PMID- 12743572 TI - Oral administration of specific antigens to allergy-prone infant dogs induces IL 10 and TGF-beta expression and prevents allergy in adult life. AB - BACKGROUND: Oral administration of allergens can induce immune tolerance to specific allergens in rodents and hence might be a possibility to prevent and treat allergic diseases in human subjects. However, the gastrointestinal tract of mice is different from that of human subjects. The absorption of specific antigens and subsequent antigen presentation to intestinal T cells is different in both species, making it difficult to extrapolate results. OBJECTIVE: We investigated primary oral tolerance to ovalbumin (OVA) in an IgE high-responder dog model, which is more predictive for human allergic diseases than corresponding rodent models. METHODS: Oral tolerance was induced by means of a 28 day treatment with OVA dissolved in cow's milk. RESULTS: We observed reduced OVA specific IgE and IgG production in response to ensuing subcutaneous challenges. Allergic conjunctivitis induced by means of ocular and airway provocation was significantly reduced in tolerized animals compared with that seen in nontolerized control animals. In addition, eosinophilia and neutrophilia in bronchoalveolar lavage fluid and bronchoconstriction after airway allergen challenge were significantly suppressed in tolerized animals. Cytokine analysis by means of real-time PCR on bronchoalveloar fluid cells after allergen challenge revealed a high-level expression of IL-10 and transforming growth factor beta, predominantly in the CD14(+) population. CONCLUSION: Feeding infant beagles with OVA for 4 weeks is sufficient to prevent hallmark manifestations of asthma and allergy in adult life. The mechanism of oral tolerance involved an increased expression of IL-10 and transforming growth factor beta cytokines. PMID- 12743573 TI - Effect of GM-CSF on immune, inflammatory, and clinical responses to ragweed in a novel mouse model of mucosal sensitization. AB - BACKGROUND: Conventional models of allergic airway inflammation involve intraperitoneal administration of ovalbumin in conjunction with a chemical adjuvant (generally aluminum hydroxide) to generate allergic airways inflammation. Here we have investigated the effect of respiratory mucosal exposure to a ragweed extract in the absence of chemical adjuvant on the generation of allergic responses. OBJECTIVES: We sought to develop a mouse model of ragweed-induced allergic airway inflammation through mucosal sensitization and to investigate the role of GM-CSF in this process. METHODS: Ragweed was delivered intranasally to an airway microenvironment enriched with GM-CSF, which was achieved by means of either multiple coadministrations of recombinant GM-CSF or a single delivery of an adenoviral vector carrying the GM-CSF transgene. RESULTS: Administration of a purified ragweed extract leads to T(H)2 sensitization (and not inhalation tolerance) accompanied by mild airway inflammation, modest clinical symptoms, and moderate production of T(H)2 cytokines by splenocytes on ragweed restimulation. The administration of anti-GM-CSF antibodies in conjunction with ragweed diminished T(H)2-associated cytokine production. These responses were amplified by enriching the airway microenvironment with GM-CSF. Under these conditions, all T(H)2-associated immune-inflammatory responses, as well as the clinical responses, were considerably enhanced. To investigate the mechanism underlying these effects, we examined lung mononuclear cells by means of flow cytometry and detected a substantial expansion of antigen-presenting cells, particularly dendritic cells, as well as a substantially increased activation of these antigen-presenting cells, as demonstrated by the expression of B7 molecules, particularly B7.2. CONCLUSION: GM-CSF plays an important role in the generation of allergic immune-inflammatory responses to ragweed. PMID- 12743574 TI - Differential roles for CD4 and CD8 T cells after diisocyanate sensitization: genetic control of TH2-induced lung inflammation. AB - BACKGROUND: Exposure to diisocyanates is a major cause of occupational asthma. We previously developed a novel mouse model of diisocyanate-induced asthma involving epicutaneous sensitization to hexamethylene diisocyanate (HDI) that demonstrates many features of the human disease, including airway eosinophilia and mucus hypersecretion. OBJECTIVE: To determine what factors are critical for the development of HDI-induced airway inflammation, we investigated the strain distribution of this response and the roles of CD4(+) and CD8(+) T cells. METHODS: Mice were epicutaneously exposed to HDI and then challenged with HDI, either by means of inhalation to induce airway inflammation or on the ear to induce contact hypersensitivity (CHS). Lymph node cytokine production and serum antibodies were also measured. RESULTS: Induction of airway eosinophilia was highly dependent on the mouse strain used, with C57BL/6, A/J, CBA, C3H, and C57BL/10 mice all having significantly fewer eosinophils than BALB/c mice. HDI specific antibodies and lymph node IL-5 and IL-13 production were also diminished in non-BALB/c strains. In contrast, CHS to HDI developed in all strains tested. Studies in mice deficient in either CD4(+) or CD8(+) T cells revealed that CD4(+) T cells were critical for HDI-induced airway eosinophilia, whereas CD8(+) T cells were the major effector cells in CHS. CONCLUSION: The data suggest that, in contrast to CHS, induction of T(H)2 responses after epicutaneous exposure to diisocyanates is strongly genetically influenced. Furthermore, the lung inflammatory response to inhaled HDI appears to depend primarily on effective generation of these CD4(+) T(H)2 responses, as is the case in atopic asthma. PMID- 12743575 TI - Intolerance to nonsteroidal anti-inflammatory drugs might precede by years the onset of chronic urticaria. AB - BACKGROUND: Recent studies have found that most otherwise normal subjects with a history of acute urticaria induced by several nonsteroidal anti-inflammatory drugs (NSAIDs) show a wheal-and-flare reaction on intradermal injection of autologous serum. This phenomenon has been previously observed in patients with chronic urticaria (CU) and suggests a possible common background in CU and NSAID induced urticaria. A relationship between these 2 conditions is further suggested by the fact that up to 30% of patients with CU have a worsening of their skin disorders after the ingestion of chemically unrelated NSAIDs. OBJECTIVE: I sought to assess whether otherwise normal subjects with multiple or single NSAID intolerance show a propensity to have CU. METHODS: Two hundred eighty otherwise normal patients with an unequivocal history of acute urticaria induced by NSAIDs seen during the last 10 years were studied. On the basis of both clinical history and oral challenge tests with at least 2 alternative NSAIDs, the patients were classified as having single or multiple NSAID intolerance. All the patients were re-evaluated within the end of 2002, 1 to 10 years after the first visit, to assess the onset of CU. One hundred allergic adults without a history of CU and of drug allergy followed up for 1 to 10 years were used as control subjects. RESULTS: One hundred fifty-nine and 121 patients were finally considered as having single or multiple NSAID intolerance, respectively. At the follow-up visit, 93 (33%) of 280 patients had CU. The prevalence of CU was very similar in subjects with single or multiple NSAID intolerance (48/159 [30%] vs 45/121 [37%], respectively; P = not significant). Only 1 (1%) of 100 atopic control subjects had CU during the follow-up period (P <.001). Among single NSAID reactors, patients who had CU had a significantly higher prevalence of intolerance to aspirin than those who did not have CU (36/48 [75%] vs 41/111 [37%], P <.001), whereas the latter had a markedly higher prevalence of intolerance to pyrazolone drugs (52/111 [47%] vs 10/48 [21%], P <.01). Altogether, only 12 (15%) of 82 patients intolerant to drugs other than aspirin versus 36 (47%) of 77 aspirin reactors had CU (P <.001). CONCLUSION: NSAID intolerance might precede the onset of CU by years. Both multiple and single NSAID reactors with a history of aspirin induced urticaria seem at higher risk for CU than patients with a history of single intolerance to NSAIDs other than aspirin. PMID- 12743576 TI - Down-regulation of vasoactive intestinal polypeptide receptor expression in atopic dermatitis. AB - BACKGROUND: Receptors for vasoactive intestinal polypeptide (VIP) have recently been suggested to play a key role in immunomodulation with genetically modified mice. However, it is not known whether changes in receptor gene regulation are involved in the pathogenesis of human immune disorders. OBJECTIVE: We studied the expression of VPAC(2) in acute lesions of the human immune disease atopic dermatitis. METHODS: By using nonradioactive in situ hybridization, quantitative immunohistochemistry, RT-PCR, and gene array studies, the expression status of VPAC(2) was assessed in atopic dermatitis and control tissues and in the human mast cell line HMC-1. RESULTS: In situ hybridization and immunohistochemistry demonstrated VPAC(2) mRNA and protein expression in human mast cells surrounded by VIP positive nerve fibers. Gene array experiments and RT-PCR studies showed high levels of VPAC(2) mRNA expression in mast cells that were increased compared to other receptors such as VPAC(1) or VIP in the human mast cell line HMC-1. Stimulation of HMC-1 cells led to a downregulation of VPAC(2). Similarly, quantitative immunohistochemistry for VPAC(2) in acute atopic dermatitis lesions showed a significantly decreased VPAC(2) immunoreactivity in mast cells. CONCLUSION: The downregulation of VPAC(2) in human mast cells in acute lesions of atopic dermatitis suggests a role of this G-protein;coupled receptor in the pathophysiology of the disease. PMID- 12743577 TI - Immediate-type hypersensitivity reaction to ingestion of mycoprotein (Quorn) in a patient allergic to molds caused by acidic ribosomal protein P2. AB - BACKGROUND: Quorn is the brand name for a line of foods made with so-called "mycoprotein," which springs from the mold Fusarium venenatum. Since the introduction on the food market, there have been complaints from consumers reporting adverse gastrointestinal reactions after ingestion of mycoprotein. To date, it is not clear whether the reported symptoms are IgE-mediated. OBJECTIVE: The aim of the study was to describe for the first time a case history of an asthmatic patient with severe hypersensitivity reactions to ingested mycoprotein and to identify and characterize the potential allergen that might be responsible for this. METHODS: The sensitization pattern of the asthmatic subject was characterized, and food allergy to mycoprotein was assessed by double-blinded placebo-controlled food challenge. Afterward, specific IgE antibodies of the serum of this patient were used to screen a Fusarium culmorum cDNA expression library. The coding sequence of one enriched cDNA-clone was expressed in Escherichia coli to produce a recombinant protein that was further purified and immunologically characterized. RESULTS: The patient showed high sensitization to many known aeroallergens but apart from Quorn not to any other tested food samples. The deduced amino acid sequence of the enriched cDNA-clone (Fus c 1) showed large identity to the 60S acidic ribosomal protein P2 which is highly conserved among several species and also described as minor allergen in other mold species. The frequency of IgE reactivity of sera from F culmorum -sensitized subjects to rFus c 1 was approximately 35%. By enzyme allergosorbent test inhibition, we found 65% inhibition of mycoprotein IgE reactivity by rFus c 1. On the opposite we found reduced IgE reactivity of rFus c 1 of 68% by using mycoprotein as inhibitor. CONCLUSIONS: Sensitization to mold allergens by the respiratory tract and subsequent oral ingestion of cross-reactive proteins may lead to severe food-allergic reactions. Thus, the 60S acidic ribosomal protein P2 of F venenatum probably is the reason for the described severe hypersensitivity reactions of the patient to Quorn-mycoprotein because of its potential cross reactivity to the F culmorum allergen Fus c 1. PMID- 12743578 TI - Penicillin skin testing in advance of need: multiyear follow-up in 568 test result-negative subjects exposed to oral penicillins. AB - BACKGROUND: There are few published data on adverse drug reactions and/or resensitization associated with oral penicillin use in penicillin allergy history positive/penicillin skin test-negative individuals during routine clinical care with multiyear follow-up. OBJECTIVES: We sought to provide long-term follow-up data on the type, severity, and frequency of adverse reactions associated with oral penicillin use in individuals who have histories of penicillin "allergy" yet also have negative results on penicillin skin tests done in advance of need. We also aimed to repeat testing on individuals with penicillin-associated adverse reactions. METHODS: Medical records were reviewed for penicillin use and associated adverse reactions in all 568 penicillin skin test-negative individuals who had received at least 1 course of oral penicillin after testing but before December 31, 2001, during routine care. These individuals were drawn from a group of 1246 penicillin skin test-negative individuals seen initially between November 16, 1994, and August 13, 2001. RESULTS: The mean length of follow-up was 4.26 +/- 1.64 years (range, 0.39-7.12 years). The mean penicillin exposure was 3.94 +/- 3.91 courses (range, 1-22 courses). Only 65 (11.4%) of 568 subjects had any penicillin-associated reactions, and 6 subjects had 2 reactions each. A reaction occurred in 27 subjects (4.8%) with their first penicillin reexposure. There were 71 (3.2%) reactions with 2236 total penicillin courses. There were no serious reactions. Repeated testing was done in 33 subjects older than age 18 years. Only 1 subject was positive on repeated penicillin skin testing. CONCLUSION: Penicillin use after negative penicillin skin testing done in advance of need is safe, and resensitization is rare. PMID- 12743579 TI - Biochemical and clinical evidence that aspirin-intolerant asthmatic subjects tolerate the cyclooxygenase 2-selective analgetic drug celecoxib. AB - BACKGROUND: Subjects with aspirin-intolerant asthma (AIA) respond with bronchoconstriction and extrapulmonary adverse reactions to conventional nonsteroidal anti-inflammatory drugs (NSAIDs) that inhibit the cyclooxygenase (COX) step in the biosynthesis of prostaglandins. Recently, 2 isotypes of COX have been identified, and COX-2-selective NSAIDs have been developed for treatment of inflammatory disorders. OBJECTIVE: We investigated whether 33 subjects with a typical history of AIA tolerated the new COX-2-selective NSAID celecoxib. METHODS: All subjects displayed current aspirin sensitivity in oral or inhalation challenge tests. The subjects first underwent a double-blind, randomized, cross-over, increasing-dose challenge with placebo or celecoxib (10, 30, or 100 mg in suspension) on 2 occasions 7 days apart. Thereafter, all subjects were exposed to 400 mg of celecoxib administered during an open challenge session as two 200-mg doses 2 hours apart. Lung function, clinical symptoms, and urinary excretion of leukotriene E(4) (LTE(4)) were monitored, with the latter being a sensitive biochemical marker of aspirin intolerance. RESULTS: There were no changes in lung function or extrapulmonary symptoms during the double-blind sessions or in urinary excretion of LTE(4). Also, the highest recommended daily dose of celecoxib was well tolerated, with no symptoms, lung function changes, or alterations in urinary LTE(4) levels. CONCLUSIONS: A group of subjects with clinically well-documented AIA tolerated acute challenge with the selective COX-2 inhibitor celecoxib. The findings indicate that the intolerance reaction in AIA is due to inhibition of COX-1. Large long-term studies of COX-2 inhibitors in AIA should be undertaken. PMID- 12743580 TI - Genetic susceptibility to food allergy is linked to differential TH2-TH1 responses in C3H/HeJ and BALB/c mice. AB - BACKGROUND: Although food allergy is a serious health problem in westernized countries, factors influencing the development of food allergy are largely unknown. Appropriate murine models of food allergy would be useful in understanding the mechanisms underlying food allergy in human subjects. OBJECTIVE: We sought to determine the susceptibility of different strains of mice to food hypersensitivity. METHODS: C3H/HeJ and BALB/c mice were sensitized to cow's milk (CM) or peanut by means of intragastric administration, with cholera toxin as a mucosal adjuvant. Mice were then challenged with CM or peanut. Antigen specific IgE levels, anaphylactic symptoms, plasma histamine levels, and splenocyte cytokine profiles of these 2 strains were compared. RESULTS: CM specific IgE levels were significantly increased only in the C3H/HeJ strain, 87% of which exhibited systemic anaphylactic reactions accompanied by significantly increased plasma histamine levels in response to challenge. BALB/c mice exhibited no significant CM-specific IgE response, increased plasma histamine levels, or anaphylactic symptoms. After peanut challenge, 100% of peanut-sensitized C3H/HeJ mice exhibited high levels of peanut-specific IgE and anaphylactic symptoms. In contrast, no hypersensitivity reactions were detected in BALB/c mice, despite the presence of significant serum peanut-specific IgE levels. Splenocytes from CM- and peanut-sensitized C3H/HeJ mice exhibited significantly increased IL-4 and IL 10 secretion, whereas splenocytes from BALB/c mice exhibited significantly increased IFN-gamma secretion. CONCLUSION: Induction of food-induced hypersensitivity reactions in mice is strain dependent, with C3H/HeJ mice being susceptible and BALB/c mice being resistant. This strain-dependent susceptibility to food allergy is associated with differential T(H)2-T(H)1 responses after intragastric food allergen sensitization. PMID- 12743581 TI - Association of chronic urticaria with thyroid autoimmunity and Raynaud phenomenon with anticentromere antibodies. PMID- 12743582 TI - IgE to Staphylococcus aureus enterotoxins in serum is related to severity of asthma. PMID- 12743583 TI - Role of profilin and polcalcin in chenopod pollen allergy. PMID- 12743584 TI - A possible role for cyclooxygenase 2 inhibitors in the treatment of chronic urticaria. PMID- 12743585 TI - Intravenous heparin did not prevent exacerbations of hereditary angioedema in a patient on maintenance hemodialysis. PMID- 12743587 TI - Antifungal nasal washes for chronic rhinosinusitis: what's therapeutic-the wash or the antifungal? PMID- 12743589 TI - FAHF-1 purporting to block peanut-induced anaphylaxis. PMID- 12743591 TI - Noon and Freeman on prophylactic inoculation against hay fever. PMID- 12743593 TI - Elidel (pimecrolimus) cream 1%: a nonsteroidal topical agent for the treatment of atopic dermatitis. AB - Elidel is a steroid-free cream containing a 1% strength of the topical immunomodulator pimecrolimus. Elidel was specifically developed as a treatment for atopic dermatitis (AD) and is approved for use in children as young as 2 years of age. The production of inflammatory cytokines by activated T cells in skin is thought to play an important role in the pathogenesis of AD. Elidel potently suppresses cytokine production by dermal T cells without significantly impairing systemic immune responses. Elidel does not cause steroid-associated local effects, such as dermal atrophy, striae, or telangiectasia. In randomized controlled clinical studies, twice-daily application of Elidel was shown to significantly improve the signs and symptoms of AD in infants, children, and adults. The clinical effect of Elidel on pruritus, the most troublesome symptom of AD, can be observed within 1 week of therapy and is maintained for the duration of treatment. Elidel is well tolerated; the risk of application-site reactions, such as itching or burning, is comparable with that of the vehicle. Adverse effects were generally mild in patients receiving Elidel and occurred at rates comparable with those in patients receiving vehicle treatment. In a 1-year study, Elidel significantly reduced the incidence of flares when used at the first signs and symptoms of acute AD. As a result, overall corticosteroid use to treat flares was significantly lower in patients using Elidel for early intervention. PMID- 12743595 TI - Identification of cJun-responsive genes in Rat-1a cells using multiple techniques: increased expression of stathmin is necessary for cJun-mediated anchorage-independent growth. AB - cJun is a major component of the transcription factor AP-1 and mediates a diverse set of biologic properties including proliferation, differentiation, and apoptosis. To identify cJun-responsive genes, we inducibly expressed cJun in Rat 1a cells and observed two distinct phenotypes: changes in cellular morphology with adherent growth and anchorage-independent growth. The biologic effects of cJun were entirely reversible demonstrating that they require the continued presence of cJun. To determine the genes, which mediate the biologic effects of cJun, we employed multiple methods including differential gene analysis, suppression subtractive hybridization, and cDNA microarrays. We identified 38 cJun-responsive genes including three uncharacterized genes under adherent and/or nonadherent conditions. Half of the known 36 genes were cytoskeleton- and adhesion-related genes, suggesting a major role of cJun in the regulation of the genes related to cell morphology. As proof of the principle that this approach could identify genes whose upregulation was necessary for nonadherent growth, we investigated one gene, stathmin whose upregulation by cJun was observed only under these conditions. Although overexpression of stathmin did not result in nonadherent growth, inhibition of stathmin protein expression by antisense oligonucleotides in cJun-induced Rat-1a cells prevented nonadherent growth. These results suggest that stathmin plays an essential role in anchorage-independent growth by cJun and may be a potential target for specific inhibitors for AP-1 dependent processes involved in carcinogenesis. PMID- 12743594 TI - EAPII interacts with ETS1 and modulates its transcriptional function. AB - Ets proteins constitute a family of conserved sequence-specific DNA-binding proteins and function as transcription factors. ETS1 plays important roles in differentiation, lymphoid cell development, invasiveness and angiogenesis. Such diverse roles of ETS1 are likely to be dependent on its associated proteins. A yeast two-hybrid screen was conducted and here we describe a novel ETS1 interacting protein designated as ETS1-associated protein II (EAPII). EAPII protein interacts with ETS1 and other Ets proteins (ETS2 and FLI1) both in vitro and in vivo. Indirect immunofluorescence demonstrated that EAPII is predominately localized to the nucleus of mammalian cells. EAPII negatively modulates ETS1 transcriptional activity and attenuates synergistic transactivation by ETS1 and AP-1. Significantly, re-expression of EAPII inhibits the migration of epithelial cancer cells, but does not affect cell viability. Therefore, EAPII is a novel ETS1 modulator that regulates specific aspects of the ETS1 functions. PMID- 12743596 TI - TMEFF1 and brain tumors. AB - TMEFF1 is a novel transmembrane protein, containing two follisatin domains and an epidermal growth factor-like region. These structural domains suggest a role for TMEFF1 in growth factor signaling. TMEFF1 fused to enhanced green fluorescent protein revealed that TMEFF1 is expressed on the cell membrane. Northern analysis of normal human tissue showed that TMEFF1 is predominantly expressed in the brain. Study of cancer cell lines from different tissues including the brain, demonstrated moderate to low levels of TMEFF1 in most of these transformed cell lines. Furthermore, quantitative real-time RT-PCR analysis of 54 brain tumors showed that most of these tumors (96%) had lower levels of TMEFF1 expression than normal brain tissue. Interestingly, ectopic expression of TMEFF1 in brain cancer cells resulted in their growth inhibition. These data suggest that TMEFF1 may behave as a tumor suppressor gene in brain cancers. PMID- 12743597 TI - Loss of von Hippel-Lindau protein causes cell density dependent deregulation of CyclinD1 expression through hypoxia-inducible factor. AB - Loss of the von Hippel-Lindau gene (VHL) expression ca-uses deregulation of contact inhibition of cell growth, which might be one of the bases of the tumor suppressor function of VHL. Here we show that this function of the VHL gene product (pVHL) depends on cell autonomous events. To identify the target gene of pVHL, which is directly involved in the contact inhibition, we compared the gene expression profile between VHL-deficient renal carcinoma 786-O cells and those infected with an adenovirus vector encoding VHL. In addition to known pVHL regulated genes, such as vascular endothelial growth factor and carbonic anhydrase, we found cyclinD1 as a new target of pVHL at a high cell density. In VHL-expressing cells (VHL (+) cells), the cyclinD1 mRNA expression level diminishes at a high cell density, while it remains at a relatively high level in VHL-deficient cells (VHL (-) cells). The cyclinD1 expression level was also abnormally high in VHL (-) cells at a high cell density. Consequently, the phosporylation level of the retinoblastoma (Rb) protein remained high in these cells, whereas there was no phosporylated Rb in VHL (+) cells under the contact inhibition. The abnormal expression of cyclinD1 at a high cell density was observed even in VHL (+) cells under the hypoxic state. Moreover, ectopic expression of a HIF mutant resistant to pVHL-mediated proteolysis causes the abnormal cyclinD1 expression in VHL (+) cells. Taken together, these observations indicate that VHL is required for the downregulation of cyclinD1 at a high cell density through HIF. PMID- 12743598 TI - High-level expression of cutaneous fatty acid-binding protein in prostatic carcinomas and its effect on tumorigenicity. AB - The expression of cutaneous fatty acid-binding protein (C-FABP) in prostate tissues was examined by immunohistochemistry. Among the 76 cases, all seven (100%) normal tissues were unstained. Of the 35 benign prostatic hyperplasia (BPH), 25 (71.4%) specimens were unstained and 10 (28.6%) were stained positively. For the 34 prostatic carcinomas, the C-FABP expression was remarkably increased: 25 (73.5%) samples stained positively, and only nine (26.5%) were unstained. Transfection of a vector expressing an antisense C-FABP transcript into the PC-3M prostatic cancer cells yielded two transfectant lines: PC-3M-CFABP 1 and PC-3M-CFABP-3, producing, respectively, a 3.8- and a 6.9-fold reduction in C-FABP levels. Comparing with the control transfectants, the in vitro invasiveness of both PC-3M-CFABP-1 and PC-3M-CFABP-3 was significantly reduced. When tested in nude mouse, the average size of tumours produced by PC-3M-CFABP-1 and by PC-3M-CFABP-3 was reduced by 2.9- and 4.2-fold respectively, in comparison with that of tumours produced by the control transfectants. Analysis showed that the decreased vascular endothelial growth factor (VEGF) and microvessel densities in the tumours were associated with the reduced C-FABP. These data show that C FABP is increased in prostatic carcinoma cells and suppression of its expression can significantly inhibit the tumorigenicity, probably by reducing the expression of VEGF. PMID- 12743599 TI - Long-term high-dose interferon-alpha therapy delays Hepadnavirus-related hepatocarcinogenesis in X/myc transgenic mice. AB - The role of interferon-alpha (IFN-alpha) remains unclear in prevention of virus induced hepatocellular carcinoma in humans. We have investigated it herewith in the X/myc transgenic mouse model of Hepadnavirus-related hepatocarcinogenesis because of upregulation of c-myc oncogene in the liver. We have demonstrated that IFN-alpha can downregulate dose-dependently hepatocyte proliferation and c-myc overexpression at early premalignant stages, while it does not affect either hepatocyte apoptosis or telomerase activity at these steps. However, continuous and long-term administration of IFN-alpha dose-dependently delays tumor onset in dysplastic livers and increases overall survival of animals, more efficiently whether started before the onset of dysplasia. The present study therefore highlights that early preventive administration of IFN-alpha can slow down evolution towards hepatocellular carcinoma via repression of c-myc and hepatocyte proliferation at premalignant steps in experimental c-myc-induced hepatocarcinogenesis. However, the transient effect observed in this study emphasizes a need to clarify the possible mechanisms of acquired resistance and subsequent therapeutic escape. Our experimental model may be a pertinent tool to explore antioncogenic properties of IFN-alpha in human cirrhotic livers showing c myc upregulation. PMID- 12743600 TI - Random mutagenesis of PDZ(Omi) domain and selection of mutants that specifically bind the Myc proto-oncogene and induce apoptosis. AB - Omi is a mammalian serine protease that is localized in the mitochondria and released to the cytoplasm in response to apoptotic stimuli. Omi induces cell death in a caspase-dependent manner by interacting with the X-chromosome linked inhibitor of apoptosis protein, as well as in a caspase-independent way that relies on its proteolytic activity. Omi is synthesized as a precursor polypeptide and is processed to an active serine protease with a unique PDZ domain. PDZ domains recognize the extreme carboxyl terminus of target proteins. Internal peptides that are able to fold into a beta-finger are also reported to bind some PDZ domains. Using a modified yeast two-hybrid system, PDZ(Omi) mutants were isolated by their ability to bind the carboxyl terminus of human Myc oncoprotein in yeast as well as in mammalian cells. One such PDZ(m) domain (PDZ-M1), when transfected into mammalian cells, was able to bind to endogenous Myc protein and induce cell death. PDZ-M1-induced apoptosis was entirely dependent on the presence of Myc protein and was not observed when c-myc null fibroblasts were used. Our studies indicate that the PDZ domain of Omi can provide a prototype that could easily be exploited to target specifically and inactivate oncogenes by binding to their unique carboxyl terminus. PMID- 12743601 TI - Nuclear localization of Y-box factor YB1 requires wild-type p53. AB - Nuclear localization and high levels of the Y-box binding protein YB1 appear to be important indicators of drug resistance and tumor prognosis. YB1 also interacts with the p53 tumor suppressor protein. In this paper, we explore a role for p53 in the nuclear localization of YB1. We report that various genotoxic stresses induce nuclear localization of YB1 in a small proportion of treated cells, but only in cells with wild-type p53. We go on to show directly that functional p53 is required for YB1 to translocate to the nucleus. Tumor associated p53 mutants however are attenuated for YB1 nuclear localization as are mutants mutated in the proline-rich domain of p53. These data link the DNA-damage response of p53 to YB1 nuclear translocation. In addition, we find that YB1 inhibits p53-induced cell death and its ability to trans-activate promoters of genes involved in cell death signaling. Together these data suggest that some forms of p53 cause YB1 to accumulate in the nucleus, which in turn inhibits p53 activity. These results provide a possible explanation for the correlation of nuclear YB1 with drug resistance and poor prognosis in some tumor types, and for the first time implicate p53 in the process of nuclear translocation. PMID- 12743602 TI - Loss of FADD protein expression results in a biased Fas-signaling pathway and correlates with the development of tumoral status in thyroid follicular cells. AB - Downregulation of proapoptotic molecules like Fas or caspase 8, or upregulation of antiapoptotic molecules like FLICE inhibitory protein has been suggested to be a regulatory mechanism set up by tumor cells to block the death signal received via death receptors. In an in-depth study of the Fas/FasL-signaling pathway in thyroid tumor development, we have demonstrated that tumor cells specifically downregulate the multideath receptor adapter Fas-associated death domain (FADD). The regulation of FADD expression occurred only at the protein level. Furthermore, in the absence of FADD, Fas-signaling resulted in accelerated growth of thyrocytes. Since thyrocytes also acquired FasL expression during tumor development, the absence of FADD protein could lead to greater resistance to numerous death receptor-mediated apoptosis, stimulation of their own proliferation through Fas/FasL interaction, and the capacity to counter-attack the infiltrating lymphocytes. PMID- 12743603 TI - Caspase inhibition switches doxorubicin-induced apoptosis to senescence. AB - The inhibition of apoptosis is generally believed to be a major determinant of resistance to chemotherapy. However, recent findings have shown that caspase inhibitors do not protect cancer cells from death by cytotoxic agents, but may switch drug-induced apoptosis to an alternative 'default death'. The primary goals of this study were to determine the major characteristics of the 'default death' and the mechanism by which this switch is activated. For this purpose, we first investigated putative cell death modes induced by doxorubicin. Molecular markers associated with these death modes were utilized to identify the default death resulting from the inhibition of apoptosis. Our findings demonstrated that doxorubicin induced at least three distinct types of cell death, senescence, apoptosis and a type of necrosis, which were concentration dependent. Specific molecular markers such as p21/WAF1, activated caspase-3 and activated Akt were associated with these death modes. The pan-caspase inhibitor (Q-VD-OPH) greatly reduced doxorubicin-induced caspase-3 activation but did not protect cells against drug toxicity. The combination of doxorubicin and Q-VD-OPH caused an increased expression of p21/WAF1 and senescence -associated -beta-galactosidase activity, but did not alter Akt activation. Collectively, these findings suggest that the inhibition of apoptosis may lead to an increased expression of cell cycle inhibitors and cellular senescence. PMID- 12743604 TI - Loss of PTEN/MMAC1/TEP in EGF receptor-expressing tumor cells counteracts the antitumor action of EGFR tyrosine kinase inhibitors. AB - We have examined the possible mechanisms of resistance to the epidermal growth factor receptor (EGFR) inhibitors in tumor cells with variable levels of EGFR. ZD1839 (Iressa) is a small-molecular-weight, ATP-mimetic that specifically inhibits the EGFR tyrosine kinase. A431 cell growth was markedly inhibited by ZD1839 (IC(50)< or =0.1 microM) whereas the MDA-468 cells were relatively resistant (IC(50)2 microM). Low doses of ZD1839 delayed cell cycle progression and induced apoptosis in A431 cells but not in MDA-468 cells. In both cell lines, 0.1 microM ZD1839 eliminated EGFR phosphorylation. However, the basal activity of the phosphatidylinositol-3 kinase (PI3 K) target Akt was eliminated in A431 but not in MDA-468 cells, implying that their Akt activity is independent of EGFR signals. A431 cells express PTEN/MMAC1/TEP, a phosphatase that can dephosphorylate position D3 of phosphatidylinositol-3,4,5 trisphosphate, the site that recruits the plecstrin-homology domain of Akt to the cell membrane. On the contrary, MDA-468 cells lack the phosphatase and tensin homolog (PTEN), potentially setting Akt activity at a high threshold that is unresponsive to EGFR inhibition alone. Therefore, we reintroduced (PTEN) by retroviral infection in MDA-468 cells. In MDA-468/PTEN but not in vector controls, treatment with ZD1839 inhibited P-Akt levels, induced relocalization of the Forkhead factor FKHRL1 to the cell nucleus, and increased FKHRL1-dependent transcriptional activity. ZD1839 induced a greater degree of apoptosis and cell cycle delay in PTEN-reconstituted than in control cells. These data suggest that loss of PTEN, by permitting a high level of Akt activity independent of receptor tyrosine kinase inputs, can temporally dissociate the inhibition of the EGFR with that of Akt induced by EGFR inhibitors. Thus, in EGFR-expressing tumor cells with concomitant amplification(s) of PI3K-Akt signaling, combined blockade of the EGFR tyrosine kinase and Akt should be considered as a therapeutic approach. PMID- 12743605 TI - SKIP3, a novel Drosophila tribbles ortholog, is overexpressed in human tumors and is regulated by hypoxia. AB - Regions of hypoxia are a hallmark of solid tumors. Tumor cells modulate the regulation of specific genes allowing adaptation and survival in the harsh hypoxic environment. We have identified SKIP3, a novel human kinase-like gene, which is overexpressed in multiple human tumors and is regulated by hypoxia. SKIP3 is an ortholog of the Drosophila tribbles, rat NIPK, dog C5FW, and human C8FW genes. Drosophila tribbles is involved in slowing cell-cycle progression during Drosophila development, but little is known regarding the function or tissue distribution of the vertebrate orthologs. We show that the normal tissue expression of SKIP3 is confined to human liver, while multiple primary human lung, colon, and breast tumors express high levels of SKIP3 transcript. Endogenous SKIP3 protein accumulates within 48 h under hypoxic growth conditions in HT-29 and PC-3 cells, with upregulation of the SKIP3 mRNA transcript by 72 h. We identified activating transcription factor 4 (ATF4) as a SKIP3-binding partner using the yeast-two-hybrid assay. Coexpression of SKIP3 and ATF4 showed that SKIP3 is associated with the proteolysis of ATF4, which can be blocked using a proteosome inhibitor. These results indicate that SKIP3 may be an important participant in tumor cell growth. PMID- 12743606 TI - The adenovirus E1A oncoprotein recruits the cellular TRRAP/GCN5 histone acetyltransferase complex. AB - The adenovirus E1A oncoprotein stimulates cell growth and inhibits differentiation by deregulating the normal transcription program via interaction with positive and negative cellular effectors. E1A associates with transcriptional regulatory complexes containing p400 and TRRAP involved in chromatin remodeling and decondensation. TRRAP is a component of three distinct human histone acetyltransferase (HAT) complexes: the TIP60 complex and complexes containing GCN5 or PCAF. We demonstrate here that E1A binds a TRRAP complex that contains the GCN5 acetyltransferase during a normal adenovirus infection. E1A binds GCN5 and TRRAP in vivo early after virus infection. E1A is associated with significant HAT activity in vitro that is partly attributable to GCN5. E1A represses c-Myc- and E2F-1-directed transcriptional activation in vivo by sequestering GCN5 and/or TRRAP. Our results demonstrate that E1A distinctly binds TRRAP/GCN5, p300/CBP and PCAF HAT complexes. Through interactions with multiple HAT complexes, E1A may deregulate cellular transcription programs and facilitate infection by recruiting functional HAT coactivators to viral and cellular promoter regions. PMID- 12743607 TI - E mu/S mu transposition into Myc is sometimes a precursor for T(12;15) translocation in mouse B cells. AB - Misguided immunoglobulin (Ig) class switch recombination (CSR) has been implicated in the origin of Myc-activating chromosomal translocations, T(12;15), in BALB/c mouse plasmacytomas (PCTs). CSR has also been involved in the progression of T(12;15); for example, the approximation of Myc to the 3'-C alpha enhancer. This study provides evidence for an additional mechanism by which aberrant CSR may facilitate T(12;15): transposition of Ig heavy-chain (IgH) sequences to Myc. Five IgH transposons containing the intronic heavy-chain enhancer, E mu, and a truncated switch mu region, S mu, were found in the first intron of Myc in lymph node cells of IL-6 transgenic BALB/c mice. In two cases E mu/S mu transposition primed Myc to get involved in apparent trans-chromosomal CSR to C gamma 1, presumably leading to T(12;15). Translocations preceded by E mu/S mu transposition can sometimes be distinguished from de novo translocations by molecular fingerprints in translocation breakpoint regions (Ig switch region [S] inversions and unusual gene orders in composite S regions). The presence of such fingerprints in some PCTs suggests that the tumors sometimes evolve from transposition-bearing precursors. We propose that E mu/S mu transposition to Myc may facilitate plasmacytomagenesis by sensitizing Myc to undergo T(12;15) translocation. T(12;15), in turn, juxtaposes Myc to the 3'-C alpha enhancer, which appears to be required for deregulating Myc in a manner that is conducive to PCT development. PMID- 12743608 TI - Fusion of an AF4-related gene, LAF4, to MLL in childhood acute lymphoblastic leukemia with t(2;11)(q11;q23). AB - We showed that the LAF4 gene on 2q11.2-12 was fused to the MLL gene on 11q23 in a pediatric patient with CD10 positive acute lymphoblastic leukemia (ALL) having t(2;11)(q11;q23). The LAF4 gene, which encodes a lymphoid nuclear protein of 1227 amino acids with transactivation potential, is thought to have a role in early lymphoid development. The LAF4 protein was homologous to AF4 and AF5q31 proteins that are fused to MLL in infant early pre-B ALL and the breakpoint of LAF4 was located within the region homologous to the transactivation domain of AF4 and AF5q31. Expression of the 8.5-kb LAF4 transcript was detected in the adult heart, brain, and placenta and in the fetal brain. LAF4 expression was found to be higher in ALL cell lines than in AML and Epstein-Barr virus-transformed B lymphocyte cell lines. These findings suggest that LAF4, AF4 and AF5q31 might define a new family particularly involved in the pathogenesis of 11q23-associated ALL. PMID- 12743611 TI - Household exposure factors, asthma, and school absenteeism in a predominantly Hispanic community. AB - The Passaic Asthma Reduction Effort (PARE) used an asthma symptom and household exposure factor questionnaire to screen 4634 elementary school children over a 4 year period in Passaic, New Jersey. During the first year, an additional 240 preschool children were also screened. Overall, 16% of the school children were reported by their parents to have been diagnosed with asthma. In all, 30% of responding families claimed to have at least one family member diagnosed with asthma and this was five times more likely if the target child had asthma. Exposures consistently associated with childhood asthma diagnosis included environmental tobacco smoke (ETS), presence of dampness/mold, roaches, and furry pets in the home. Diagnosis of asthma was primarily associated with all six symptoms used in the PARE questionnaire, and secondarily with environmental factors. Puerto Rican and black children had the highest asthma prevalence (26% and 33%), while Mexican children had the lowest (7%). Use of medications and school absenteeism among asthmatic children were associated with wheeze and night cough, but not with any specific environmental exposure. Increased school absenteeism by children undiagnosed with asthma was associated with ETS and dampness/mold in the home. Differences in asthma diagnosis and absenteeism in response to environmental factors were found across ethnic subgroups. Getting asthmatic children on medical management protocols and providing families with education about environmental risk reduction should aid in reducing morbidity in this ethnically complex population. Such coordinated efforts offer the promise of reducing school absenteeism. PMID- 12743612 TI - A national survey of persistent, bioaccumulative, and toxic (PBT) pollutants in the United States milk supply. AB - This study measured 21 persistent, bioaccumulative, and toxic (PBT) pollutants in the US milk supply. Since milk fat is likely to be among the highest dietary sources of exposure to PBTs, it is important to understand their levels in this food. Nationwide samples were collected from 45 dairy plants in July of 2000 and again in January 2001. The levels of all chemicals in the chlorobenzene, pesticide and other halogenated organic groups were determined to be below their detection limits in all samples. National averages were computed for 11 chemicals or chemical groups found above the detection limits. The national average CDD/CDF and PCB TEQ concentrations were 14.30 and 8.64 pg/l, respectively, for a total of 22.94 pg/l. These levels are about half the values found in a similar study conducted in 1996. If this difference is in fact indicative of declining milk levels and assuming exposure levels from nondairy pathways have remained the same over this time period, this would result in an overall decrease in adult background dioxin exposure of 14%. Six PAHs were detected with national averages ranging from 40 to 777 ng/l. Cadmium concentrations ranged from 150 to 870 ng/l with a national average of 360 ng/l. Lead concentrations were consistently higher than those of cadmium, ranging from 630 to 1950 ng/l with a national average of 830 ng/l. PAHs showed the strongest seasonal/geographic differences, with higher levels in winter than summer, north than south and east than west. Average adult daily intakes from total milk fat ingestion were computed for all detected compounds and compared to total intakes from all pathways: CDD/CDF/PCB TEQs: 8 vs. 55 pg/day, PAHs: 0.6 vs. 3 micro g/day, lead: 0.14 vs. 4-6 micro g/day, and cadmium: 0.06 vs. 30 micro g/day. PMID- 12743613 TI - Aggregate exposures of nine preschool children to persistent organic pollutants at day care and at home. AB - In the summer of 1997, we measured the aggregate exposures of nine preschool children, aged 2-5 years, to a suite of organic pesticides and other persistent organic pollutants that are commonly found in the home and school environment. The children attended either of two child day care centers in the Raleigh-Durham Chapel Hill area of North Carolina and were in day care at least 25 h/week. Over a 48-h period, we sampled indoor and outdoor air, play area soil and floor dust, as well as duplicate diets, hand surface wipes, and urine for each child at day care and at home. Our target analytes were several polycyclic aromatic hydrocarbons (PAH), organochlorine pesticides, and polychlorinated biphenyls (PCB); two organophosphate pesticides (chlorpyrifos and diazinon), the lawn herbicide 2,4-dichlorophenoxyacetic acid (2,4-D), three phenols (pentachlorophenol (PCP), nonyl phenols, and bisphenol-A), 3,5,6-trichloro-2 pyridinol (TCP), and two phthalate esters (benzylbutyl and dibutyl phthalate). In urine, our target analytes were hydroxy-PAH, TCP, 2,4-D, and PCP. To allow estimation of each child's aggregate exposures over the 48-h sampling period, we also used time-activity diaries, which were filled out by each child's teacher at day care and the parent or other primary caregiver at home. In addition, we collected detailed household information that related to potential sources of exposure, such as pesticide use or smoking habits, through questionnaires and field observation. We found that the indoor exposures were greater than those outdoors, that exposures at day care and at home were of similar magnitudes, and that diet contributed greatly to the exposures. The children's potential aggregate doses, calculated from our data, were generally well below established reference doses (RfDs) for those compounds for which RfDs are available. PMID- 12743614 TI - Acephate exposure and decontamination on tobacco harvesters' hands. AB - Agricultural workers manually harvesting tobacco have the potential for high dermal fexposure to pesticides, particularly on the hands. Often gloves are not worn as it hinders the harvesters' ability to harvest the tobacco leaves. To enable harvesters to remove pesticide residue on the hands and decrease absorbed doses, the EPA Worker Protection Standard requires growers to have hand-wash stations available in the field. The purpose of this study was to measure the concentration of acephate residue on the hands of tobacco harvesters, and the effectiveness of hand washing in reducing the acephate residue. Hand-wipes from the hands of 12 tobacco harvesters were collected at the end of the morning and at the end of the afternoon over 2 consecutive days. Each harvester had one hand wiped prior to washing his hands, and the other hand-wiped after washing his hands with soap and water. In addition to the hand-wipe samples, leaf-wipe samples were collected from 15 tobacco plants to determine the amount of acephate residue on the plants. The average acephate level in leaf-wipe samples was 1.4 ng/cm(2). The geometric mean prewash and postwash acephate levels on the hands were 10.5 and 0.4 ng/cm(2), respectively. Both prewash (P-value=0.0009) and postwash hand (P-value=0.01) samples were positively correlated with leaf-wipe concentrations. Tobacco harvester position tended to influence hand exposure. Hand washing significantly reduced acephate levels on the hand, after adjusting for sampling period, hand sampled, job position, and leaf-wipe concentration (P value< or =0.0001) with levels reduced by 96%. A substantial amount of acephate was transferred to the hands, and while hand washing significantly reduced the amount of residue on the hands, not all residue was removed. PMID- 12743615 TI - House dust and inorganic urinary arsenic in two Arizona mining towns. AB - Residents of copper mining and smelting towns may have increased risk of arsenic exposure from elevated arsenic contained in environmental media. To determine the relation of arsenic in house dust to inorganic urinary arsenic concentrations, a door-to-door survey was conducted in Hayden and Winkelman, Arizona. A total of 122 households (404 individuals) participated; 85 provided dust samples. Urine was collected at first morning void and analyzed for total and speciated arsenic. Speciation of arsenic was performed in samples with total arsenic above 10 micro g/l (N=106). The generalized estimating equation was used to determine the relation between urinary and house dust arsenic concentrations, allowing adjustment for the correlation of measurements obtained from the same home. Seafood consumption during the past 3 days and smoking contributed significantly to inorganic urinary arsenic, after adjusting for age and gender. Arsenic in house dust was not significantly associated with inorganic urinary arsenic measurements in this population. PMID- 12743616 TI - Exposure of infants to outdoor and indoor air pollution in low-income urban areas - a case study of Delhi. AB - Indoor air pollution is potentially a very serious environmental and public health problem in India. In poor communities, with the continuing trend in biofuel combustion coupled with deteriorating housing conditions, the problem will remain for some time to come. While to some extent the problem has been studied in rural areas, there is a dearth of reliable data and knowledge about the situation in urban slum areas. The microenvironmental model was used for assessing daily-integrated exposure of infants and women to respirable suspended particulates (RSP) in two slums of Delhi - one in an area of high outdoor pollution and the other in a less polluted area. The study confirmed that indoor concentrations of RSP during cooking in kerosene-using houses are lesser than that in wood-using houses. However, the exposure due to cooking was not significantly different across the two groups. This was because, perhaps due to socioeconomic reasons, kerosene-using women were found to cook for longer durations, cook inside more often, and that infants in such houses stayed in the kitchen for longer durations. It was observed that indoor background levels during the day and at nighttime can be exceedingly high. We speculate that this may have been due to resuspension of dust, infiltration, unknown sources, or a combination of these factors. The outdoor RSP levels measured just outside the houses (near ambient) were not correlated with indoor background levels and were higher than those reported by the ambient air quality monitoring network at the corresponding stations. More importantly, the outdoor levels measured in this study not only underestimated the daily-integrated exposure, but were also poorly correlated with it. PMID- 12743617 TI - Identifying US populations for the study of health effects related to drinking water arsenic. AB - The US Environmental Protection Agency recently set a new maximum contaminant level (MCL) for arsenic in drinking water of 10 micro g/l. In this paper, we review the completeness and accuracy of drinking water arsenic occurrence data in the United States and identify populations exposed to elevated arsenic concentrations that would be suitable for epidemiological studies of arsenic health effects. Using existing data from the Environmental Protection Agency Arsenic Occurrence and Exposure Database and additional data from state health and environment departments and water utilities, we identified 33 counties in 11 states with an estimated mean drinking water arsenic concentration of 10 micro g/l or greater. A total of 11 of these 'confirmed' counties had an estimated mean arsenic concentration of 20 micro g/l or more and two had an estimated mean arsenic concentration 50 micro g/l or more. Based on census data, between 1950 and 1999 there were approximately 51.1 million person-years of exposure to drinking water arsenic at levels of 10 micro g/l or more, 8.2 million at levels of 20 micro g/l or more arsenic and 0.9 million at levels of 50 micro g/l or more. Mortality and incidence of diseases known to be associated with arsenic exposure can and should be examined in these counties as part of a comprehensive assessment of arsenic health effects in US populations. PMID- 12743618 TI - Traffic density in California: socioeconomic and ethnic differences among potentially exposed children. AB - Motor vehicles are the main source of many hazardous air pollutants in California. Previous studies have shown that low-income and minority populations are more likely to live near industrial sources of pollution and in areas that do not meet national air quality standards. We estimated neighborhood exposures to motor vehicle emissions from a road network with daily traffic counts using a geographic information system. To calculate traffic density, we summed the average daily vehicle miles of travel per square mile of land area for each census block group in the state. We used 1990 census data to characterize the population by age, race and socioeconomic status in block groups with high traffic density. Block groups with more than 500,000 vehicle miles of travel per square mile were defined to be high traffic density. Statewide, about 5% of all block groups met this criterion and more than 215,000 children under 15 years of age lived in these high traffic density areas. Block groups in the lowest quartile of median family income were three times more likely to have high traffic density than block groups in the highest income quartile. The percentage of children living in high traffic density block groups increased with decreasing median family income for all race and ethnicities except White. Overall, children of color were about three times more likely to live in high-traffic areas than were white children. Based on this analysis, low-income and children of color have higher potential exposure to vehicle emissions. Future exposure assessment studies should target the highest traffic density areas, and health studies should consider the differences by income and race or ethnicity during design. PMID- 12743619 TI - Comments on the NMDA antagonist memantine. PMID- 12743620 TI - A model with original conditions and materials to test potential inhibitors of cyclooxygenase-2 versus cyclooxygenase-1 in isolated human blood cells. AB - The inflammatory process can be influenced by an enzyme known as cyclooxygenase, which exists in two isoforms: COX-1, the constitutive form, and COX-2, which is only synthesized in the case of inflammation and has negative side effects. To decrease these undesirable effects, current research seeks to discover compounds that selectively inhibit COX-2 versus COX-1. When added to the samples, arachidonic acid activates the synthesis of COX-2. This explains why the two isoforms proceed in different ways. As for COX-1, human lymphocytes and monocytes were activated with 50 microM of calcium ionophore for 1 h at 37 degrees C with 50 microM of arachidonic acid, and the quantity of thromboxane B(2) (TxB(2)) was measured by radioimmuno-assay (RIA). As for COX-2, cells were first incubated with acetylsalicylic acid to block COX-1, then stimulation of COX-2 was performed using 8 mcg/ml lipopolysaccharide (LPS) for 3 h at 37 degrees C with 50 microM arachidonic acid. The IC(50) values for reference compounds were reproducible, taking into consideration the passage through the plasmatic membrane and the transduction signal, while avoiding the problems of binding with plasma proteins and using identical cells, unlike other models. PMID- 12743621 TI - CCK-8 modulates GABA-stimulated [3H]-flunitrazepam binding in vitro. AB - In membranes from rat cerebral cortex, cholecystokinin-8 (CCK-8) did not modulate basal [(3)H]-flunitrazepam binding at either 4 degrees C or 37 degrees C. At a concentration of 10(-6) M, CCK(-8) significantly decreased gamma-amino-butyric acid (GABA)-stimulated (10(-6) M) [(3)H]-flunitrazepam binding at 37 degrees C. Scatchard analyses suggest that the decreased GABA-stimulated binding might be due to a decrease in the affinity of benzodiazepine receptors rather than to a decrease of number of binding sites. The observed modulation of benzodiazepine receptors by CCK-8 in vitro might explain some of the functional interactions between CCK and benzodiazepine systems. PMID- 12743622 TI - Inmunoferon, an immunomodulator of natural origin, does not affect the rat liver cytochrome P-450 and phase II conjugation enzymes. AB - Inmunoferon is a glycoconjugate of natural origin with immunomodulatory properties. It has recently been shown to regulate TNF-alpha expression induced by lipopolysaccharide (LPS) challenge through a hypothalamo-pituitary-adrenal (HPA)-dependent mechanism. Inmunoferon is orally administered to immunocompromised patients as an adjuvant during immune therapy such as vaccination or infectious diseases treatment. Due to its mainly adjuvant nature, it is necessary to determine if coadministration of Inmunoferon affects the activity of other drugs. In this study we analyzed the possible modification of the hepatic drug biotransformation system by using Inmunoferon in a rat model, which may result in changes in the biological activity of other drugs administered simultaneously. Inmunoferon-treated animals showed no differences to control littermates in antipyrine metabolism. No differences were found in either cytochrome P-450 and b5 levels or cytochrome P-450-dependent activities and phase II conjugation enzymes in lysates from Inmunoferon-treated rat hepatic cells. The same treatment reduced levels of serum TNF-alpha in LPS-challenged animals. In summary, Inmunoferon is unable to affect the hepatic bioconjugation system during administration and thus seems unlikely to interact with, or modify the effect of, coadministered drugs. PMID- 12743623 TI - Effects of [Arg8]-vasopressin on regional cerebral blood flow in spontaneously hypertensive rats. AB - The effects of [Arg(8)]-vasopressin (AVP) and related compounds on regional cerebral blood flow (rCBF) in the hippocampus were studied using conscious spontaneously hypertensive rats (SHR). rCBF in the hippocampus decreased gradually with age in proportion to an increase in mean blood pressure. Subcutaneous injection of AVP caused a dose-dependent increase in rCBF in the hippocampus. The effects of the metabolic fragments AVP4-9 and AVP4-8 on rCBF were relatively weak. OPC-31260, a vasopressin V(2) antagonist, antagonized the AVP-induced increase in rCBF in the hippocampus. Furthermore, subcutaneous injection of DDAVP, a V(2) agonist, increased rCBF in the hippocampus. On the other hand, the AVP-induced increase in rCBF in the hippocampus was not antagonized by OPC-21268, a vasopressin V(1) antagonist. Intracerebroventricular injection of AVP caused no significant changes in rCBF in the hippocampus, even at a dose of 10 ng/site. PMID- 12743624 TI - Laboratory evaluation of the hypoglycemic effect of Anacardium occidentale Linn (Anacardiaceae) stem-bark extracts in rats. AB - This study evaluated the hypoglycemic effect of stem-bark extracts of Anacardium occidentale Linn., of the Anacardiaceae family, in normal (normoglycemic) and in streptozotocin-treated diabetic rats. Young adult, male Wistar rats weighing 250 300 g were used. Diabetes mellitus was induced in the test rats by intraperitoneal injections of streptozotocin (STZ, 90 mg/kg). In one set of experiments, graded doses of the aqueous and methanolic stem-bark extracts of A. occidentale (100-800 mg/kg p.o.) were separately administered to groups of fasted normal and fasted diabetic rats. In another set of experiments, 800 mg/kg p.o. of the aqueous or methanolic extract of the plant, a dose which produced maximal hypoglycemic effects in both fasted normal and diabetic rats in the previous set of experiments, were used. The hypoglycemic effects of single doses (i.e., 800 mg/kg p.o.) of A. occidentale stem-bark aqueous and methanolic extracts were compared with those of insulin (5 microU/kg s.c.) and glibenclamide (0.2 mg/kg p.o.) in both fasted normal and fasted diabetic rats. Following acute treatment, relatively moderate-to-high doses of A. occidentale stem-bark extracts (100-800 mg/kg p.o.) produced dose-dependent, significant reductions (p< 0.05-0.001) in the blood glucose concentrations of both fasted normal and fasted diabetic rats. On their own, both insulin (5 microU/kg s.c.) and glibenclamide (0.2 mg/kg p.o.) produced significant reductions (p< 0.01-0.001) in the blood glucose concentrations of the fasted normal and fasted diabetic rats. At single doses of 800 mg/kg p.o., A. occidentale stem-bark aqueous and methanolic extracts significantly reduced (p< 0.001) the mean basal blood glucose concentrations of fasted normal and fasted diabetic rats. The hypoglycemic effect of the methanolic plant extract was found to be slightly more pronounced than that of the aqueous plant extract in both the normal and diabetic rats examined. A. occidentale contains a diverse group of chemical compounds. Since methanol extractives of plants usually contain many chemical compounds, each of which is capable of producing definite biological activities via different mechanisms, it is difficult to draw any logical conclusion on the mechanism of the hypoglycemic effect of such a diverse mixture of chemical compounds contained in the plant extracts used in this study. While it is possible that the hypoglycemic effects of the plant extracts may be due, at least in part, to their terpenoid and/or coumarin contents, the mechanism of their hypoglycemic action remains largely speculative. However, this is unlikely to be due to the stimulation of pancreatic beta-cells and subsequent secretion of insulin. Although A. occidentale stem-bark aqueous or methanolic extract is less potent than insulin as an antidiabetic agent, the results of this experimental animal study indicate that it possesses hypoglycemic activity, and thus lends credence to the folkloric use of the plant in the management and/or control of adult-onset, type-2 diabetes mellitus among the Yoruba-speaking people of Western Nigeria. PMID- 12743625 TI - Protective effect of pioglitazone against multiple low-dose streptozotocin induced diabetes in rats. AB - The autoimmune process is one of the etiological factors of diabetes in humans. Thiazolidinediones, which act through peroxisome proliferator-activated receptor gamma, have been recently used to prevent diabetic-associated complications in patients with diabetes and insulin resistance. In the present study, we investigated the effect of pioglitazone on the diabetes induced by multiple low dose streptozotocin (MLDS) in rats. When Sprague-Dawley rats were injected intraperitoneally with a sub-diabetogenic dose of streptozotocin (STZ; 40 mg/kg/day) for a period of 5 days, they developed hyperglycemia 2 days after posttreatment. Pioglitazone (6 mg/kg) administered orally for 7 days before the first dose of STZ prevented or delayed the development of MLDS-induced diabetes compared with the group treated only with STZ. Pioglitazone treatment showed no effect on plasma glucose levels in the control group. These findings suggest that pioglitazone prevented the autoimmune process involved in the development of MLDS induced diabetes by decreasing glucose levels in rats. PMID- 12743627 TI - The antiangiogenic and therapeutic implications of endostatin. AB - Angiogenesis plays a vital role in the pathology of cancer, ischemic diseases and chronic inflammation, among other conditions. Endostatin, a newly found protein that is distributed in some parts of the human body, has been demonstrated to have a strong inhibitory role in angiogenesis. It specifically inhibits the proliferation of endothelial cells and induces their apoptosis both in vitro and in vivo. Preclinical research has proven its effective role in the treatment of various experimental tumors in rodents. Although endostatin therapy has entered phase II clinical trials in the USA, the exact mechanism and its effects on antiangiogenesis, especially the action on the suppression of endothelial cell proliferation and induction of apoptosis, remain unclear. The treatment modality for malignancies and other angiogenesis-related diseases still requires further analysis. PMID- 12743628 TI - Gateways to clinical trials. AB - Gateways to clinical trials is a guide to the most recent clinical trials in current literature and congresses. The data in the following tables has been retrieved from the Clinical Studies knowledge area of Prous Science Integrity, the drug discovery and development portal, http://integrity.prous.com. This issue focuses on the following selection of drugs: 5A8; Agomelatine, alefacept, almotriptan, anakinra, APC-8015, atazanavir, atomoxetine hydrochloride, azimilide hydrochloride; Bicifadine; Cannabidiol, caspofungin acetate, CAT-213, CGP-51901, ciclesonide, cipamfylline; Darbepoetin alfa, desloratadine, dibotermin alfa, DX 9065a; Ecogramostim, efalizumab, eletriptan, eniluracil, EPI-KAL2, erlosamide, ertapenem sodium, etilevodopa, etoricoxib, ezetimibe; Fosamprenavir calcium, fosamprenavir sodium, fumagillin; Gadofosveset sodium, gefitinib, gemtuzumab ozogamicin; HSPPC-96, human papillomavirus vaccine; Icatibant Id-KLH, imatinib mesylate, INS-37217, iodine (I131) tositumomab; LAS-34475, levobupivacaine hydrochloride, levocetirizine, linezolid, 131I-lipiodol, lonafarnib, lopinavir, LY-450108; Magnetites, MBI-594AN, melagatran, melatonin, mepolizumab, mycophenolic acid sodium salt; NC-100100; 1-Octanol, omalizumab, omapatrilat, onercept; PEG-filgrastim, (PE)HRG21, peginterferon alfa-2a, peginterferon alfa 2b, pleconaril, pneumococcal 7-valent conjugate vaccine, prasterone; Ranelic acid distrontium salt, rasagiline mesilate, reslizumab, rFGF-2, rhOP-1, rosuvastatin calcium, roxifiban acetate; Sitaxsentan sodium, sodium lauryl sulfate; Tadalafil, telithromycin, tenofovir disoproxil fumarate, tipranavir, TMC-114, tucaresol; Valdecoxib, voriconazole; Ximelagatran; Zofenopril calcium, zosuquidar trihydrochloride. PMID- 12743626 TI - Effect of amlodipine and hormone replacement therapy on blood pressure and bone markers in menopause. AB - The aim of this study was to observe the effect of an 8-week treatment with amlodipine, alone or in combination with hormone replacement therapy (HRT), on blood pressure (BP), serum osteocalcin, bone-specific alkaline phosphatase (B ALP) and urine deoxypiridinoline in postmenopausal osteoporotic women with mild to-moderate arterial hypertension. Both conventional clinical BP measurements and ambulatory blood pressure monitoring (ABPM) were used. Twenty hypertensive menopausal women with osteoporosis were randomly divided in two groups according to the treatment regimens: amlodipine and amlodipine + HRT. Neither treatment regimen significantly changed bone formation or bone resorption markers. There were no significant differences in levels of serum and urinary calcium and phosphorous or serum cholesterol and low-density lipoprotein (LDL)-cholesterol after treatment with amlodipine alone or in combination with HRT. Triglycerides were significantly decreased and high-density lipoprotein (HDL)-cholesterol was significantly increased after amlodipine treatment. Both treatment regimens significantly decreased conventionally measured BP to a similar extent. Amlodipine given alone lowered the midline estimating statistic of rhythm (MESOR; mean 24-level) of systolic BP and induced phase advances of the circadian rhythms of systolic, diastolic and mean BP. When combined with HRT, amlodipine lowered the MESOR and reduced the amplitude of systolic BP without any phase change. In conclusion, amlodipine is effective in reducing BP in postmenopausal women. The maintenance of a normal circadian BP pattern is also influenced by supplementation with 17beta-estradiol. The 8-week treatment with amlodipine alone and in combination with HRT is not associated with a marked influence on bone metabolism. PMID- 12743629 TI - Meeting the challenge of chronic care. AB - Although chronic care is the predominant mode of illness in developed countries, medical care systems are still organized as if practice addresses acute illness. Major changes are needed in the way medical care is delivered. Emphasis on brief encounters (be they hospital admissions or clinic visits) must transform into a focus on the entire episode of an illness. Patients must become more actively involved in monitoring and attending to their illnesses. Monitoring systems should emphasize specific salient elements of a given chronic disease and compare actual and expected clinical courses. Management should be based on early deviations from the expected course. The roles of both professional and nonprofessional practitioners should be reexamined to look for opportunities for role delegation; teamwork should be used sparingly. The linkage between medical care and long-term care requires shared goals that reflect a concern with slowing the rate of decline in care recipients. (c) 2001 Prous Science. All rights reserved. PMID- 12743630 TI - Pitavastatin (NK-104), a new HMG-CoA reductase inhibitor. AB - Pitavastatin calcium (NK-104) is a new synthetic hydroxymethylglutaryl-coenzyme A (HMG-CoA) reductase inhibitor (statin). Animal studies suggest that, in addition to reducing low-density lipoprotein (LDL) cholesterol, the drug may produce marked reductions in triglyceride-rich particles (very-low-density [VLDL] and intermediate-density lipoproteins [IDL]). It is not metabolized by the common cytochrome P-450 3A4 enzyme, possibly reducing the risk for drug interactions. Early studies suggest that it may be quite useful for treating common dyslipidemias (isolated elevations of LDL cholesterol and combined disorders with elevations of LDL cholesterol and triglycerides). Such improvements in lipid profiles are proven to have positive effects on cardiovascular risk. Human studies are under way to further elucidate the effects of the drug and procure approval by various regulatory bodies. (c) 2001 Prous Science. All rights reserved. PMID- 12743631 TI - The role of telomerase in joint deterioration in rheumatoid arthritis. AB - The rate of bone formation is largely determined by the number of osteoblasts, which in turn is determined by the rate of replication of progenitors and the life span of mature cells, reflecting the timing of death by apoptosis. To date, however, the exact age-dependent changes of the cellular activity, replicative potential and life span of osteoblasts have not been investigated. Here we review evidence that the cellular activity, telomere lengths and replicative life span of osteoblastic cells obtained from juxtaarticular bone marrow gradually decrease with the advance of donor age in arthritides. Also, we show that the ability to extend the cellular life span of osteoblasts may have important implications for biological research and the development of new technologies for bone and joint diseases. (c) 2001 Prous Science. All rights reserved. PMID- 12743632 TI - New serum tests for the diagnosis of prostate cancer. AB - Prostate-specific antigen (PSA) has had a profound impact on the early diagnosis, treatment and follow-up of prostate cancer, the most common malignancy in men. However, it is not only a marker for prostate cancer but is also often expressed in benign conditions such as benign prostatic hyperplasia, prostatitis and other inflammatory disorders. For early detection of prostate cancer, limitations of the use of PSA become obvious; its widespread use has led to extensive expensive and often unnecessary diagnostic procedures associated with significant morbidity. New serum tests derived from total PSA may play a key role in enhancing the accuracy of prostate cancer diagnosis. The ratio of free to total PSA improves specificity while maintaining a high sensitivity for prostate cancer detection for men with a total PSA of 2.5-10 ng/ml who also have a normal digital rectal examination. Human glandular kallikrein 2 also has the potential to be a valuable tool in combination with both total and free PSA for the early diagnosis of prostate cancer. However, the optimal clinical use of human glandular kallikrein 2 still remains to be clarified. Complex PSA seems to be a reliable tool and equivalent alternative to total PSA to improve specificity at high sensitivity levels in men with suspected prostate cancer, mainly for PSA levels below 4 ng/ml. Several newly discovered isoforms of free PSA (bPSA, [-2]pPSA and inactive intact PSA) may also impact the early detection of prostate cancer, with encouraging preliminary results that warrant further clinical investigation. (c) 2001 Prous Science. All rights reserved. PMID- 12743633 TI - Telithromycin (HMR 3647): The first ketolide antibiotic. AB - Telithromycin, the first ketolide antibiotic to undergo clinical development, has been specifically designed to treat community-acquired respiratory tract infections, including those caused by resistant pathogens. Like macrolides, telithromycin inhibits protein synthesis by acting mainly on the 50S ribosomal subunit. The defining structural characteristic is a keto function in place of the C3-cladinose moiety, which greatly improves acid stability and confers a lack of induction of MLSb resistance. Telithromycin provides potent activity against a wide spectrum of Gram-positive and Gram-negative organisms, including erythromycin-resistant pneumococci and atypical/intracellular organisms. Preliminary results from clinical trials have demonstrated that telithromycin may provide a convenient and effective compact treatment option for select respiratory tract infections such as community-acquired pneumonia, acute bacteria exacerbations of chronic bronchitis, acute sinusitis and tonsillitis/pharyngitis. (c) 2001 Prous Science. All rights reserved. PMID- 12743634 TI - IL-12-Based therapy of malignancies. AB - Interleukin-12 (IL-12) is an important molecule that triggers the activation of natural killer (NK) cells and T-cells, the development of T helper type 1 (Th1) cells and the expression of antiangiogenic genes. Novel methods for IL-12 delivery include cell-based ex vivo gene therapy, viral vector-based gene therapy and DNA plasmid-based nonviral gene therapy. IL-12 electroporation gene therapy may hold some promise for tumors accessible by electrode, such as head and neck cancer, breast cancer, prostate cancer and melanoma. Codelivery of other therapeutic genes with IL-12 may enhance the therapeutic effect and reduce the level of IL-12 required for efficacy. All three approaches to IL-12 gene therapy are under clinical investigation. The preliminary results indicate that IL-12 gene therapy is safe and is not associated with any major clinical toxicity. (c) 2001 Prous Science. All rights reserved. PMID- 12743635 TI - The role of letrozole (Femara(R)) in breast cancer therapy: A clinical review. AB - Letrozole is a third-generation aromatase inhibitor for use in postmenopausal women with hormonal-sensitive breast cancer. This drug was found to reduce or effectively shrink tumors in a significant number of such patients. It exhibits antitumor activity at a relatively low daily dose, and is highly potent and selective and well tolerated. Results from recent phase III clinical studies have confirmed the efficacy and the key role of this drug in the therapy of advanced breast cancer in postmenopausal women. Moreover, letrozole demonstrated higher activity and lower toxicity compared to tamoxifen in the first-line therapy of postmenopausal women affected with advanced breast cancer. However, it also represents a valid option in second-line therapy after tamoxifen failure. New data on this agent in adjuvant or neoadjuvant treatment also suggest efficacy in the treatment of early breast cancer. This article reviews the clinical data on letrozole in all settings and its future potential in chemoprevention. (c) 2001 Prous Science. All rights reserved. PMID- 12743636 TI - Clinical trials with glycoprotein IIb/IIIa antagonists - No benefit without bleeding? AB - As the glycoprotein GPIIb/IIIa receptor is the final common pathway in platelet aggregation, antagonists of this receptor cause a profound inhibition of aggregation induced by any agonist. The short-term efficacy and safety of GPIIb/IIIa antagonists in patients undergoing coronary angioplasty was demonstrated with murine 7E3 Fab, but this antibody was immunogenic. Abciximab is a chimeric human-mouse monoclonal antibody that is less immunogenic. The first major trial with a GPIIb/IIIa antagonist was the EPIC trial with abciximab, which showed that abciximab reduced the ischemic complications of coronary balloon angioplasty and atherectomy in high-risk patients, but increased the risk of bleeding. Subsequent studies showed that using less concurrent heparin reduced bleeding. Abciximab also reduced the rate of revascularization. Further studies have shown that the benefits of abciximab extended to all patients undergoing angioplasty (EPILOG), including patients with unstable angina (CAPTURE) and acute myocardial infarction (RAPPORT). Clinical trials with eptifibatide and tirofiban have failed to demonstrate benefit, at the doses used, in angioplasty. Abciximab and eptifibatide, but not oral xemilofiban, improve the safety of the coronary stenting procedure. Short-term intravenous treatment with lamifiban, eptifibatide or tirofiban is beneficial in acute coronary syndromes (unstable angina, non-Q wave myocardial infarction). Orally active GPIIb/IIIa antagonists are being developed for use in acute coronary syndromes and myocardial infarction. However, no benefit has been shown with lefradafiban in acute coronary syndromes and sibrafiban and orbofiban are harmful. Eptifibatide, lamifiban and abciximab improve coronary patency in myocardial infarction, and long- term trials of GPIIb/IIIa antagonists are being conducted in acute myocardial infarction. Abciximab can cause thrombocytopenia, and all the GPIIb/ IIIa antagonists increase the incidence of bleeding, but there is no excess of intracranial hemorrhage. (c) 2001 Prous Science. All rights reserved. PMID- 12743637 TI - Imaging of the gastrointestinal tract. AB - Gastrointestinal imaging has undergone immense and profound changes since the days when radiologists had only plain abdominal radiography, barium studies and cholecystography to offer as a means of investigating the gastrointestinal tract. Advances in technology have made huge differences in the ways and means of investigating the various pathologies afflicting the gastrointestinal tract. Ultrasound provides assessment of the solid organs and the intestine without the use of ionizing radiation. Computed tomography (CT) provides unparalleled imaging of congenital, inflammatory, traumatic and neoplastic processes, providing information on the solid organs and tubular structures, including the vasculature. Information is provided to aid immediate management or to stage lesions for subsequent or combined therapies. Magnetic resonance imaging (MRI) gives unprecedented multiplanar images and soft tissue resolution. There is still a place for plain radiography and barium studies but this is decreasing as technology advances.(c) 2001 Prous Science. All rights reserved. PMID- 12743638 TI - Pharmacodynamics of drug-induced weight gain. AB - Body weight gain during treatment with drugs for any kind of disease may represent improvement of the disease itself. However, sometimes these drug induced alterations of the body's appetite-regulating mechanisms result in excessive weight gain, thus jeopardizing compliance with prescribed medication. A number of drugs are capable of changing body weight as an adverse consequence of their therapeutic effect. Included in this category are the psychotropic drugs such as antipsychotics, antidepressants and mood stabilizers. Antipsychotics are well-known culprits of weight gain. The low-potency (e.g., chlorpromazine and thioridazine) and atypical agents (e.g., clozapine, olanzapine, quetiapine and risperidone) are most often associated with weight gain. Antidepressants such as tricyclic antidepressants and monoamine oxidase (MAO) inhibitors are most often associated with significant weight gain. The tertiary tricyclic antidepressant amitriptyline is thought to induce the most weight gain. Mood stabilizers such as lithium carbonate, valproic acid and carbamazepine also induce weight gain in a considerable number of patients. Treatment with corticosteroids is associated with dose-dependent body weight gain in many patients and corticosteroid-induced obesity aggravates other corticosteroid-associated health risks. Insulin therapy in diabetic patients usually increases body weight. Finally, sulfonylurea derivatives, antineoplastic agents used for the treatment of breast cancer and several drugs used in migraine prophylaxis may cause body weight gain as well. (c) 2001 Prous Science. All rights reserved. PMID- 12743639 TI - Prevention of delayed cerebral vasospasm after aneurysmal subarachnoid hemorrhage. AB - The delayed cerebral vasoconstriction known as cerebral vasospasm remains a significant cause of permanent neurological deficit and death following aneurysmal subarachnoid hemorrhage despite the best current medical therapies. The mechanism of cerebral vasospasm remains unknown. Several new drugs have been tested in animal models of subarachnoid hemorrhage, and these experimental studies have contributed to a better understanding of the potential mechanisms that lead to cerebral vasospasm. In this article, the authors highlight recent advances in the various treatment procedures for delayed cerebral vasospasm following subarachnoid hemorrhage. (c) 2001 Prous Science. All rights reserved. PMID- 12743640 TI - Ezetimibe. AB - Ezetimibe is the first of a new class of drugs that specifically reduces the intestinal absorption of cholesterol. The drug is absorbed into the intestinal epithelial cell and remains associated in great part with the apical cell membrane where it is believed to interfere with the putative sterol transporter system. This apparently prevents both free cholesterol and plant sterols (phytosterols) from being transported into the cell from the intestinal lumen. The mechanism is very different from the reduction produced by phytosterol esters and phytostanol esters that have been documented previously as interfering with the micellar presentation of sterols to the cell surface. The drug is rapidly absorbed and glucuronidated in the intestinal cell before secretion into the blood. Ezetimibe is avidly taken up by the liver from the portal blood and excreted into the bile, resulting in low peripheral blood concentrations. The glucuronide conjugate is hydrolyzed and absorbed and is equally effective in inhibiting sterol absorption. This enterohepatic recycling is responsible for a half-life in the body of more than 20 h. The principle medical benefit appears to be a reduction in low-density lipoprotein cholesterol (LDLc). There is also a reduction in the cholesterol content of chylomicrons, which may provide some reduction in the atherogenic potential of the plasma lipoprotein pool. Triglycerides fall moderately and a modest rise in high-density lipoprotein cholesterol (HDLc) has been consistently observed in groups of treated patients. The maximum mean reduction of LDL cholesterol is approximately 20-25% in small studies at the maximal dose tested of 40 mg/day and the reduction is usually in the 16-20% range at the dosage of 10 mg/day. Most subsequent studies have been completed with the 10 mg/day dose. The medication is effective as a single daily dose due to its long residence time in the body. Combinations of ezetimibe with all available statins have been completed and demonstrate LDLc reductions of approximately 25% as additive effects to any statin dose alone. There are also small additional increases in HDL (2-3%) and reductions in triglycerides (10 15%). Additive effects to statin therapy have also been documented in patients with homozygous familial hypercholesterolemia. Reduction of plant sterols has been demonstrated in phytosterolemia, offering the first drug treatment for this rare inherited disease. PMID- 12743641 TI - Clopidogrel in acute coronary syndromes (unstable angina and non-Q-wave myocardial infarction). AB - Given the importance of thrombosis in acute coronary syndromes, antithrombotic therapy has become standard treatment for these conditions. This article reviews the mechanism of action and the major evidence supporting the clinical use of clopidogrel, a potent antiplatelet agent of the thienopyridines class, focusing on its role in the setting of acute coronary syndromes without persistent ST segment elevation (unstable angina and non-Q wave myocardial infarction). Some unanswered questions relating to this medication are also highlighted. Finally, current updates on clinical guidelines for the use of clopidogrel in acute coronary syndromes are discussed. PMID- 12743642 TI - Telomerase as a promising target for human cancer gene therapy. AB - A number of different approaches have been developed to target human cancer on the basis of its specific expression of telomerase activity. The most common approach relies on inhibition of telomerase activity for reversion of the immortal phenotype of tumor cells. Different components and types of inhibitors targeting various regulatory levels have been regarded as useful for telomerase inhibition. Most methods, however, rely on successive telomere shortening. This process is very slow and causes a long time lag between the onset of inhibition and the occurrence of senescence or apoptosis as a reversal of the immortal phenotype. Many telomerase inhibitors seem to be most efficient when combined with conventional chemotherapeutics. There are some promising approaches to circumvent the slow track of telomere shortening and induce fast apoptosis in treated tumor cells. It has been demonstrated that telomerase may be involved in triggering apoptosis, but the underlying molecular mechanism remains unclear. Other important strategies are the use of telomerase promoters for the application of toxic or pro-apoptotic drugs into human cancer cells or the use of immunological properties of the telomerase enzyme for possible cancer therapies. PMID- 12743643 TI - Aminoglycoside ototoxicity. AB - Over the last 10 years new information has been published providing a better understanding of the risk factors, mechanism and prevention of aminoglycoside ototoxicity. The use of a higher dose and once-daily intravenous administration of aminoglycosides has shown clinical effectiveness with no increase in ototoxicity when compared to traditional regimens. An enhanced susceptibility to aminoglycoside cochlear toxicity has been linked to an A-to-G substitution in location 1555 of the mitochondrial ribosomal ribonucleic acid (RNA). More recently, a second mutation involving a thymidine deletion in the 12s ribosomal RNA gene has been identified which can predispose patients to aminoglycoside auditory toxicity. Experimental evidence in animals has indicated that reactive oxygen species are one of the most important factors responsible for the development of aminoglycoside ototoxicity. The animal data has suggested a decrease in hearing loss induced by aminoglycosides when antioxidant or iron chelator therapy is given concomitantly with aminoglycoside antibiotics. PMID- 12743645 TI - Combining medical informatics and bioinformatics toward tools for personalized medicine. AB - OBJECTIVES: Key bioinformatics and medical informatics research areas need to be identified to advance knowledge and understanding of disease risk factors and molecular disease pathology in the 21 st century toward new diagnoses, prognoses, and treatments. METHODS: Three high-impact informatics areas are identified: predictive medicine (to identify significant correlations within clinical data using statistical and artificial intelligence methods), along with pathway informatics and cellular simulations (that combine biological knowledge with advanced informatics to elucidate molecular disease pathology). RESULTS: Initial predictive models have been developed for a pilot study in Huntington's disease. An initial bioinformatics platform has been developed for the reconstruction and analysis of pathways, and work has begun on pathway simulation. CONCLUSIONS: A bioinformatics research program has been established at GE Global Research Center as an important technology toward next generation medical diagnostics. We anticipate that 21 st century medical research will be a combination of informatics tools with traditional biology wet lab research, and that this will translate to increased use of informatics techniques in the clinic. PMID- 12743644 TI - The role of uncoupling protein 2 in the development of type 2 diabetes. AB - Uncoupling proteins (UCP) are carriers expressed in the mitochondrial inner membrane that uncouple oxygen consumption by the respiratory chain from ATP synthesis. UCP2 is a member of the multigenic UCP family that is expressed in a wide range of tissues and organs. Possible functions of UCP2 include control of ATP synthesis, regulation of fatty acid metabolism and control of reactive oxygen species production. UCP2 expression in tissues involved in lipid and energy metabolism and mapping of the gene to a region linked to obesity and hyperinsulinemia prompted studies on the involvement of UCP2 in metabolic disorders, and especially in type 2 diabetes. In human adipose tissue and skeletal muscle, UCP2 expression is increased during fasting. The carrier was shown to be under the control of fatty acids and thyroid hormones in vivo. An upregulation has been observed in the liver during high-fat feeding and obesity. However, data in UCP2 gene knockout mice do not support a role for UCP2 in steatohepatitis. The most compelling metabolic role of UCP2 comes from studies in pancreatic beta cells. Overexpression in isolated pancreatic islets results in decreased ATP content and blunted glucose-stimulated insulin secretion. UCP2 deficient mice show an increased ATP level and an enhanced insulin secretion. Lack of UCP2 dramatically improves insulin secretion and decreases hyperglycemia in leptin-deficient mice. The role of UCP2 in the control of insulin secretion constitutes, to date, the most pertinent path to investigate in a therapeutic perspective. PMID- 12743646 TI - Reflections on an arranged marriage between bioinformatics and health informatics. AB - OBJECTIVE: To compare the discussions of two workshops held during 2001 by two Canadian organisations, HEALNet, a Network of Centres of Excellence for research in health information applications, and Genome Canada, a national research funding agency for genomics and proteomics, in collaboration with the Institute of Genetics of the Canadian Institutes of Health Research, to examine strategic research development in Health Informatics and Bioinformatics respectively. METHODS: Invited workshops with structured debate. Concept analysis of preparative material and debates. RESULTS: A predominantly common set of concepts was discerned from both workshops. Analysis of published definitions showed an inability to distinguish a definition that would suggest that health informatics and bioinformatics are separate disciplines. In both workshops there was evidence of deep concerns of identity, the lack of clear structures to support research funding as well as uncertainty in distinguishing between service and research. CONCLUSIONS: Many deep issues currently inhibit the recognition and funding of research in health and bioinformatics in Canada and elsewhere. Some of these issues are common to both health and bioinformatics. The overlap in prevailing definitions, research concerns and methodological content in the respective domains suggest that common research needs should be better identified and reinforced for the benefit of both. PMID- 12743647 TI - Establishing an agenda for biomedical informatics. AB - OBJECTIVE: To describe potential areas of collaboration between Medical Informatics (BI) and Bioinformatics (BI) and their effects on planning future work in both disciplines. METHODS: Some reflections on the objectives and rationale underpinning MI and BI are given, and preliminary results from the BIOINFOMED workgroup, supported by the European Commission, are introduced. RESULTS: Applications from both subfields suggest topics for sharing and exchange between the subfields within the emerging field of Biomedical Informatics. CONCLUSIONS: We suggest how the nature and degree of collaboration between the sub-disciplines can impact future work in molecular medicine. PMID- 12743649 TI - Internet health resources: from quality to trust. AB - OBJECTIVES: Quality of online health resources remains a much debated topic, despite considerable international efforts. The lack of a systematic and comprehensive conceptual analysis is hindering further progress. Therefore we aim at clarifying the origins, nature and interrelations of pertinent concepts. Further, we claim that quality is neither a necessary nor a sufficient condition for Internet health resources to produce an effect offline. As users' trust is also required, we examine the relation of quality aspects to trust building online. METHODS: We reviewed and analyzed the key documentation and deliverables of quality initiatives, as well as relevant scientific publications. Using the insights of philosophy, we identified the elementary dimensions which underlie the key concepts and theories presented so far in the context of online health information quality. We examined the interrelations of various perspectives and explored how trust as a phenomenon relates to these dimensions of quality. RESULTS: Various aspects associated with the quality of online health resources originate from four conceptual dimensions: epistemic, ethical, economic and technological. We propose a conceptual framework that incorporates all these perspectives. We argue that total quality exists only if all four dimensions have been addressed adequately and that high total quality is conducive to warranted trust. CONCLUSIONS: Quality and trust are intertwined, but distinct concepts, and their relation is not always straightforward. Ideally, trust should track quality. Apprehending the composition of these concepts will help to understand and guide the behavior of both users and providers of online information, as well as to foster warranted trust in online resources. The framework we propose provides a conceptual starting point for further deliberations and empirical work. PMID- 12743648 TI - Informatics united: exemplary studies combining medical informatics, neuroinformatics and bioinformatics. AB - OBJECTIVES: Medical informatics, neuroinformatics and bioinformatics provide a wide spectrum of research. Here, we show the great potential of synergies between these research areas on the basis of four exemplary studies where techniques are transferred from one of the disciplines to the other. METHODS: Reviewing and analyzing exemplary and specific projects at the intersection of medical informatics, neuroinformatics, and bioinformatics from our experience in an interdisciplinary research group. RESULTS: Synergy emerges when techniques and solutions from medical informatics, bioinformatics, or neuroinformatics are successfully applied in one of the other disciplines. Synergy was found in 1. the modeling of neurophysiological systems for medical therapy development, 2. the use of image processing techniques from medical computer vision for the analysis of the dynamics of cell nuclei, and 3. the application of neuroinformatics tools for data mining in bioinformatics and as classifiers in clinical oncology. CONCLUSIONS: Each of the three different disciplines have delivered technologies that are readily applicable in the other disciplines. The mutual transfer of knowledge and techniques proved to increase efficiency and accuracy in a manifold of applications. In particular, we expect that clinical decision support systems based on techniques derived from neuro- and bioinformatics have the potential to improve medical diagnostics and will finally lead to a personalized delivery of healthcare. PMID- 12743650 TI - DataGrid, prototype of a biomedical grid. AB - BACKGROUND: The availability of large amounts of data in heterogeneous formats and the rapid progress in fields such as computer based drug design, medical imaging and medical simulations have lead to a growing demand for large computational power and easy accessibility to heterogeneous data sources. OBJECTIVES: The goal is to address these needs by deploying computing grids. Grids provide both large scale and distributed storage facilities and an increased computing power. Moreover, Grids are a promising tool to foster the synergy between bioinformatics and computerised medical imaging. METHODS: A first biomedical grid is being deployed within the framework of the DataGrid IST project (http://www.edg.org). The goal of the project is to provide a novel environment to support globally distributed scientific exploration involving up to multi-Perabyte datasets. RESULTS AND CONCLUSIONS: The first biomedical applications deployed inside the project demonstrate the relevance of the grid paradigm for genomics and medical image processing. They also highlight the specific requirements of the biomedical community. PMID- 12743651 TI - Privacy enhancing techniques - the key to secure communication and management of clinical and genomic data. AB - OBJECTIVES: To introduce some of the privacy protection problems related to genomics based medicine and to highlight the relevance of Trusted Third Parties (TTPs) and of Privacy Enhancing Techniques (PETs) in the restricted context of clinical research and statistics. METHODS: Practical approaches based on two different pseudonymisation models, both for batch and interactive data collection and exchange, are described and analysed. RESULTS AND CONCLUSIONS: The growing need of managing both clinical and genetic data raises important legal and ethical challenges. Protecting human rights in the realm of privacy, while optimising research potential and other statistical activities is a challenge that can easily be overcome with the assistance of a trust service provider offering advanced privacy enabling/enhancing solutions. As such, the use of pseudonymisation and other innovative Privacy Enhancing Techniques can unlock valuable data sources. PMID- 12743652 TI - Integr8: enhanced inter-operability of European molecular biology databases. AB - OBJECTIVES: The increasing production of molecular biology data in the post genomic era, and the proliferation of databases that store it, require the development of an integrative layer in database services to facilitate the synthesis of related information. The solution of this problem is made more difficult by the absence of universal identifiers for biological entities, and the breadth and variety of available data. METHODS: Integr8 was modelled using UML (Universal Modelling Language). Integr8 is being implemented as an n-tier system using a modern object-oriented programming language (Java). An object relational mapping tool, OJB, is being used to specify the interface between the upper layers and an underlying relational database. RESULTS: The European Bioinformatics Institute is launching the Integr8 project. Integr8 will be an automatically populated database in which we will maintain stable identifiers for biological entities, describe their relationships with each other (in accordance with the central dogma of biology), and store equivalences between identified entities in the source databases. Only core data will be stored in Integr8, with web links to the source databases providing further information. CONCLUSIONS: Integr8 will provide the integrative layer of the next generation of bioinformatics services from the EBI. Web-based interfaces will be developed to offer gene-centric views of the integrated data, presenting (where known) the links between genome, proteome and phenotype. PMID- 12743653 TI - Probabilistic graphical models for computational biomedicine. AB - BACKGROUND: As genomics becomes increasingly relevant to medicine, medical informatics and bioinformatics are gradually converging into a larger field that we call computational biomedicine. OBJECTIVES: Developing a computational framework that is common to the different disciplines that compose computational biomedicine will be a major enabler of the further development and integration of this research domain. METHODS: Probabilistic graphical models such as Hidden Markov Models, belief networks, and missing-data models together with computational methods such as dynamic programming, Expectation-Maximization, data augmentation Gibbs sampling, and the Metropolis-Hastings algorithm provide the tools for an integrated probabilistic approach to computational biomedicine. RESULTS AND CONCLUSIONS: We show how graphical models have already found a broad application in different fields composing computational biomedicine. We also indicate several challenges that lie at the interface between medical informatics, statistical genomics, and bioinformatics. We also argue that graphical models offer a unified framework making it possible to integrate in a statistically meaningful way multiple models ranging from the molecular level to cellular and to clinical levels. Because of their versatility and firm statistical underpinning, we assert that probabilistic graphical models can serve as the lingua franca for many computationally intensive approaches to biology and medicine. As such, graphical models should be a foundation of the curriculum of students in these fields. From such a foundation, students could then build towards specific computational methods in medical informatics, medical image analysis, statistical genetics, or bioinformatics while keeping the communication open between these areas. PMID- 12743654 TI - Model driven therapy - the instance of computer assisted medical interventions. AB - OBJECTIVES: Taking into account a priori knowledge is a key issue to meet the medical, scientific and industrial challenges of the progresses of Minimally Invasive Surgery. We propose an overview of these challenges. METHODS: Models play a major role in representing the relevant knowledge to plan and realize complex medical and surgical interventions. We analyze the three basic steps of Perception, Decision and Action, and illustrate by some instances how models may be integrated in these steps. RESULTS: We propose a selection of the results obtained in Model Driven Therapy. These results illustrate the issues of Perception (models allow accurate reconstruction of 3D objects from a limited set of X-ray projections), Decision (models allow to take into account elastic and dynamic characteristics of muscles), and Action (models allow to design innovative navigational and robotics aids to the realization of complex interventions). Likewise, models play a major role in the process of surgeon's education, which leads to the concept of Virtual Orthopedic University. CONCLUSIONS: Model Driven Therapy emerges as the way to perform optimal medical and surgical interventions, providing physicians and surgeons with the possibility to augment their capacities of sensing multi-modal information, of combining them to define optimal strategies, and of performing accurate and safe actions. PMID- 12743655 TI - Computational integrative physiology: at the convergence of the life and computational sciences. AB - OBJECTIVE: In this paper we outline how Computational Integrative Physiology (CIP) can help unravel the mechanisms of normal and pathological biological processes. Our objective is to illustrate how CIP is firmly grounded on the life and computational sciences. METHOD: After describing a general theoretical frame work for CIP, we will center our discussion on cardiac rhythmic disorders with a particular focus on the Long QT syndrome that will serve as a case example. Within this context, we will describe multi-scale processes in biological, medical and in general mathematical terms, starting from the control of gene expression to the electrical activity of the entire heart. We will therefore proceed from the smaller microscopic scales to the larger macroscopic ones. In doing so, we will illustrate, at least in a qualitative sense, how CIP can be accomplished by showing some of the relations that can exist between mathematical variables characterizing models of different space-scales. CONCLUSION: We will conclude by putting forth how CIP and the related fields of bioinformatics and medical informatics are necessary to derive meaningful knowledge from the huge and exponentially growing biological and medical data. PMID- 12743656 TI - Medical informatics and the quality of health: new approaches to support patient care - findings from the IMIA Yearbook of Medical Informatics 2003. AB - OBJECTIVES: The Yearbook of Medical Informatics is published annually by the International Medical Informatics Association (IMIA) and contains a selection of excellent papers on medical informatics research which have been recently published (http://www. yearbook.uni-hd.de). The 2003 Yearbook of Medical Informatics took as its theme the role of medical informatics for the quality of health care. In this paper, we will discuss challenges for health care, and the lessons learned from editing IMIA Yearbook 2003. RESULTS AND CONCLUSIONS: Modern information processing methodology and information and communication technology have strongly influenced our societies and health care. As a consequence of this, medical informatics as a discipline has taken a leading role in the further development of health care. This involves developing information systems that enhance opportunities for global access to health services and medical knowledge. Informatics methodology and technology will facilitate high quality of care in aging societies, and will decrease the possibilities of health care errors. It will also enable the dissemination of the latest medical and health information on the web to consumers and health care providers alike. The selected papers of the IMIA Yearbook 2003 present clear examples and future challenges, and they highlight how various sub-disciplines of medical informatics can contribute to this. PMID- 12743658 TI - Missense mutations of the interleukin-12 receptor beta 1(IL12RB1) and interferon gamma receptor 1 (IFNGR1) genes are not associated with susceptibility to lepromatous leprosy in Korea. AB - Interleukin-12 receptor beta 1 ( IL12RB1), interleukin-12 receptor beta 2 ( IL12RB2), and interferon gamma receptor 1 ( IFNGR1) perform important roles in the host defense against intracellular pathogens such as Mycobacteria. Several mutations within their genes have been confirmed as associated with increased susceptibility to mycobacterial infection. However, the association between mutations of the IL12RB1, IL12RB2, and IFNGR1 encoding genes and lepromatous leprosy has not been studied. This study screened for polymorphisms within IL12RB1, IL12RB2, and IFNGR1 encoding genes in the Korean populations using polymerase chain reaction (PCR)/single-strand conformation polymorphism (SSCP) DNA sequencing assay, and an association study was performed using the missense mutations of 705 A/G (Q214R), 1196 G/C (G378R), 1637 G/A (A525T), and 1664 C/T (P534S) of the IL12RB1, 83 G/A (V14M), and 1443 T/C (L467P) for the IFNGR1 encoding genes. There were no differences in the genotype and allele frequencies of IL12RB1 and IFNGR1 genes between 93 lepromatous leprosy patients and 94 control subjects. In conclusion, missense mutations of 705 A/G (Q214R), 1196 G/C (G378R), 1637 G/A (A525T), 1664 C/T (P534S) of the IL12RB1, 83 G/A (V14 M), and 1443 T/C (L467P) of the IFNGR1 encoding genes have no association with the susceptibility to lepromatous leprosy in the Korean population. PMID- 12743659 TI - Percutaneous balloon angioplasty for the treatment of iliofemoral arterial stenosis resulting from hyperhomocysteinemia in a child. AB - Angioplasty has long been used in the treatment of peripheral arterial occlusive disease with variable long-term results. In this case report, we present a patient with marfanoid features and acute ischemia of the lower extremity. Angioplasty provided us with the means of emergently relieving the acute symptoms. The patient was subsequently found on further work-up to have hyperhomocysteinemia (HC) and appropriate medical therapy was initiated. We conclude that angioplasty should be considered in pediatric patients with acute occlusive limb ischemia, providing excellent immediate results to preserve the limb as well as the time to investigate the underlying causes. PMID- 12743657 TI - Chicken MHC class I and II gene effects on antibody response kinetics in adult chickens. AB - The major histocompatibility complex (MHC) plays an important role in regulation of the immune response. The MHC class I and II genes were selected as candidates to investigate associations with vaccine response to Salmonella enteritidis and kinetics of antibody response to sheep red blood cell (SRBC) and Brucella abortus. Primary antibody response after S. enteritidis vaccination at day 10, and antibody response to SRBC and killed B. abortus after immunization at 19 and 22 weeks were measured in an F2 population. The resource population was derived from males of two highly inbred MHC-congenic Fayoumi chicken lines (M5.1 and M15.2) mated with highly inbred G-B1 Leghorn line hens. Secondary phase parameters of minimum titers ( Y(min)), maximum titers ( Y(max)), and time needed to achieve Y(min) ( t(min)) and Y(max) ( t(max)) were estimated from post secondary titers by using a non-linear regression model. Associations of single nucleotide polymorphisms (SNPs) in MHC class I and II genes with antibody response parameters were determined by a general linear model. Significant associations were found primarily in the M15.2 grandsire haplotype. There were significant associations between MHC class I alpha(1) and alpha(2) SNPs and antibody response to S. enteritidis, primary antibody response to B. abortus, Y(min) to SRBC, and Y(max) to both SRBC and B. abortus. There were significant effects of the MHC class II beta(1) domain SNP on S. enteritidis antibody and Y(max) to SRBC. The results suggest that the characterized SNPs might be used in future applications by marker-assisted selection to improve vaccine response and immunocompetence in chickens. PMID- 12743661 TI - CT scout films: don't forget to look! AB - BACKGROUND: A lateral scout view of the head is always obtained when performing head computed tomography (CT). It is common knowledge that viewing the lateral scout view may provide additional information. For a variety of reasons, however, a careful review may not be performed routinely. OBJECTIVE: To illustrate the value of the lateral scout view, we present a series of representative cases. PATIENTS: Six patients with clinically relevant findings on the scout view. RESULTS: Most of the ancillary findings were in the upper cervical spine/neck, which is typically included on the lateral scout view. CONCLUSION: Careful evaluation of the scout view of the head CT, including the skull and neck, may yield valuable information, which may not be visualized on the axial CT images. PMID- 12743660 TI - CT versus MR in neonatal brain imaging at term. AB - BACKGROUND: Recent reports have highlighted the lifetime risk of malignancy from using ionizing radiation in pediatric imaging. Computed tomography (CT), which uses ionizing radiation, is employed extensively for neonatal brain imaging of term infants. Magnetic resonance (MR) provides an alternative that does not use ionizing radiation. OBJECTIVE: The purpose of this study was to assess the cross modality agreement and interobserver agreement of CT and MR brain imaging of the term or near-term neonate. MATERIALS AND METHODS: Brain CT and MR images of 48 neonates were retrospectively reviewed by two pediatric neuroradiologists. CT and MR examinations had been obtained within 72 h of one another in all patients. CT was obtained with 5 mm collimation (KV=120, mAs=340). MR consisted of T1-weighted imaging (TR/TE=300/14; 4-mm slice thickness/1-mm gap), T2-weighted imaging (TR/TE/etl= 3000/126/16; 4-mm slice thickness/1-mm gap), and line scan diffusion imaging (LSDI) (TR/TE/b factor=1258/63/750; nominal 4-mm slice thickness/3-mm gap). The brain was categorized as normal or abnormal on both CT and MR. RESULTS: Ischemic injury was the most common brain abnormality demonstrated. McNemar's test indicated no significant difference between CT and MR test results for reader 1 (P=0.22) or reader 2 (P=0.45). The readers agreed on the presence or absence of abnormality on CT in 40 patients (83.3%) and on MR in 45 patients (93.8%). For CT, the kappa coefficient indicated excellent interobserver agreement (kappa=0.68), although the lower limit of the 95% confidence interval extends to kappa=0.55, which indicates only good-to-moderate agreement. For MR, the kappa coefficient indicated almost perfect interobserver agreement (kappa=0.88) with the 95% confidence interval extending to a lower limit of kappa=0.76, which represents excellent agreement. CONCLUSION. Because MR demonstrates findings similar to CT and has greater interobserver agreement, it appears that MR is a superior test to CT in determining brain abnormalities in the term neonate. Furthermore, since MR eliminates the use of ionizing radiation, a putative cause of malignancy, it should be the standard in neonatal brain imaging. Future efforts should be directed to improving neonatal access to MR to avoid the routine use of CT in infants. PMID- 12743662 TI - Left pulmonary artery sling presenting as unilateral echogenic lung on 20-week detailed antenatal ultrasound examination. AB - BACKGROUND: We report the first case of left pulmonary artery sling (LPAS) presenting as unilateral echogenic lung at a routine 20-week antenatal US examination. The infant had minimal symptoms at delivery, but developed an oxygen requirement and respiratory distress from day 10 of postnatal life. Antenatal US and antenatal MRI indicated an intrathoracic anomaly, so permitting definitive investigations in the neonatal period using CT, MRI and helical-CT-acquired virtual bronchoscopy. The infant successfully underwent corrective surgery. REASON TO REPORT: Antenatal ultrasound detected an anomaly permitting definitive diagnostic investigations to be performed shortly after birth. Investigations were completed before significant signs and symptoms developed. WHAT WAS UNIQUE: This is the first report of antenatally detected unilateral echogenic lung leading to the diagnosis of pulmonary artery sling. RAMIFICATIONS OF THIS REPORT: This report provides further evidence that improvements in antenatal ultrasound and MRI permit earlier diagnosis and improve patient management. PMID- 12743663 TI - Cortical laminar necrosis in brain infarcts: serial MRI. AB - High-signal cortical lesions are observed on T1-weighted images in cases of brain infarct. Histological examination has demonstrated these to be "cortical laminar necrosis", without haemorrhage or calcification. We report serial MRI in this condition in 12 patients with brain infarcts. We looked at high-signal lesions on T1-weighted images, chronological changes in signal intensity and contrast enhancement. High-signal cortical lesions began to appear about 2 weeks after the ictus, were prominent at 1-2 months, then became less evident, but occasionally remained for up to 1.5 years. They gave high signal or were isointense on T2 weighted images and did not give low signal at any stage. Contrast enhancement of these lesions was prominent at 1-2 months, and less apparent from 3 months, but was seen up to 5 months. PMID- 12743664 TI - Extensive cortical damage in a case of Creutzfeldt-Jacob disease: clinicoradiological correlations. AB - MRI demonstrated extensive cortical involvement in a patient with pathologically proven Creutzfeldt-Jacob disease. The whole brain was atrophic; some of the supratentorial cortex, putamen and caudate nucleus gave high signal on T2 weighted images; the changes were more extensive on diffusion-weighted images (DWI). Comparison of the history, and the sites of atrophy and signal change suggested that the latter predominates in regions with long-lasting damage and prevalent gliosis, while high signal on DWI indicate current neuronal loss. This case widens the range of MRI findings in patients with Creutzfeldt-Jacob disease, and suggests that some information about the progression of the disease can be extracted from single MRI study. PMID- 12743665 TI - Narrow, duplicated internal auditory canal. AB - A narrow internal auditory canal (IAC) constitutes a relative contraindication to cochlear implantation because it is associated with aplasia or hypoplasia of the vestibulocochlear nerve or its cochlear branch. We report an unusual case of a narrow, duplicated IAC, divided by a bony septum into a superior relatively large portion and an inferior stenotic portion, in which we could identify only the facial nerve. This case adds support to the association between a narrow IAC and aplasia or hypoplasia of the vestibulocochlear nerve. The normal facial nerve argues against the hypothesis that the narrow IAC is the result of a primary bony defect which inhibits the growth of the vestibulocochlear nerve. PMID- 12743666 TI - Intraorbital ophthalmic artery aneurysm. PMID- 12743668 TI - Left ventricular function and cardiovascular events following adjuvant therapy with adenosine in acute myocardial infarction treated with thrombolysis, results of the ATTenuation by Adenosine of Cardiac Complications (ATTACC) study. AB - BACKGROUND: Reperfusion therapy for acute myocardial infarction (AMI) reduces mortality but is also associated with reperfusion injury. The present study tested the hypothesis that adjuvant therapy with a low anti-inflammatory dose of adenosine might prevent reperfusion injury and preserve left ventricular function. METHODS: Six hundred and eight patients with ST-elevation AMI were randomised to receive infusions of adenosine (10 microg x kg(-1) x min(-1)) or placebo (saline) to be started with thrombolysis and maintained for 6 h. The primary endpoint was global and regional left ventricular systolic and diastolic function, as assessed by two-dimensional and Doppler echocardiography before hospital discharge. The secondary end-point was all cause and cardiovascular mortality, and non-fatal myocardial infarction during 12 months of follow-up. RESULTS: No beneficial effect of adenosine was found regarding echocardiographic indices of left ventricular systolic or diastolic function. Recruitment was stopped due to this apparent lack of effect after an interim analysis. However, after 12 months of follow-up, cardiovascular mortality was 8.9% with adenosine and 12.1% with placebo treatment [odds ratio (OR) 0.71, 95% confidence interval (C.I.) 0.4-1.2, P=0.2] among all patients and 8.4% vs 14.6% (OR 0.53, 95% C.I. 0.23-1.24, P=0.09) among patients with anterior AMI. All cause mortality differed similarly. Non-fatal AMI was not reduced similarly by adenosine treatment. Survival curves indicate that possible survival benefits are maintained after the first year of follow-up. CONCLUSIONS: Adenosine, given as adjunctive treatment with thrombolysis, did not provide detectable improvement of echocardiographic indices of left ventricular function when assessed before hospital discharge. Cardiovascular and all cause mortality appear to have been reduced by low-dose adenosine treatment, and the size of the effect appears to be clinically relevant (absolute risk reductions of approximately 4%). The power of the study regarding morbidity and mortality was, however, limited. The results are compatible with a beneficial anti-inflammatory effect of adenosine treatment on reperfusion injury after thrombolysis, which may be mediated by inhibition of leukocytes in peripheral blood. A larger trial is warranted to possibly establish beneficial effects of low-dose adenosine on survival after thrombolysis. PMID- 12743669 TI - Effects of fluticasone plus salmeterol versus twice the dose of fluticasone in asthmatic patients. AB - OBJECTIVE: Current guidelines advocate adding a long acting beta(2)-agonist (LABA) to an inhaled corticosteroid as an alternative to increasing the dose of the latter. Since it is unclear how this translates into effects on surrogate inflammatory markers, we evaluated the anti-inflammatory activity of fluticasone plus salmeterol in combination versus twice the dose of fluticasone alone. METHODS: Fifteen mild-to-moderate asthmatics (mean FEV(1) 80% predicted) uncontrolled on inhaled corticosteroids (mean dose 470 microg) were randomised in a single-blind crossover fashion to receive 2 weeks each of fluticasone 250 microg plus salmeterol 50 microg in combination (FP+SM), 1 puff b.i.d., and fluticasone 500 microg (FP), 1 puff b.i.d. Prior to each randomised treatment, there was a 2-week run-in and washout period during which patients used their usual inhaled corticosteroid therapy. Measurements were made before and after randomised treatment periods. The primary outcome was airway hyper-responsiveness to adenosine monophosphate (AMP PC(20)), while secondary endpoints were exhaled tidal nitric oxide (NO), forced expiratory volume in 1 second (FEV(1)) and forced mid-expiratory flow (FEF(25-75)). RESULTS: For AMP PC(20), FP alone but not FP+SM conferred a significant ( P<0.05) improvement amounting to 3.27 (95% CI 1.46 7.32) and 1.44 (95% CI 0.64-3.23) geometric mean fold shifts, respectively, from baseline, while the difference between treatments was significantly ( P<0.05) greater with FP alone: a 2.26-fold (95% CI 1.01-5.07) difference. Both FP alone and FP+SM conferred significant ( P<0.05) falls in NO from baseline: 2.33 (95% CI 1.71-3.19) and 1.49 (95% CI 1.09-2.03) geometric mean fold changes, respectively, while between treatments the reduction was significantly ( P<0.05) greater with FP alone: a 1.57-fold (95% CI 1.15-2.14) difference. Neither treatment significantly improved FEV(1) or FEF(25-75). CONCLUSION: Double the dose of FP alone relative to FP+SM conferred superior effects on surrogate inflammatory markers but not on lung function. Long-term studies are required to evaluate whether these improvements on surrogate inflammatory markers translate into commensurate reductions in airway remodelling and exacerbations. PMID- 12743671 TI - Cytochrome P4502E1 (CYP2E1) expression in peripheral blood lymphocytes: evaluation in hepatitis C and diabetes. AB - OBJECTIVE: Cytochrome P4502E1 (CYP2E1) is expressed in human peripheral blood lymphocytes (PBLs), and previous reports have suggested the possibility of using this readily available tissue as a reporter of CYP2E1 status. To further explore the relevance of this approach we assessed CYP2E1 expression in PBLs in two contrasting conditions, chronic hepatitis C and insulin-dependent diabetes (IDD), illustrating an organ and a systemic disease, respectively. METHODS: Total RNA was isolated from extracted PBLs (hepatitis C patients + IDD) and by percutaneous needle biopsy (hepatitis C patients only). Gene expression for CYP2E1 was determined by real-time reverse-transcription polymerase chain reaction. Histological changes in liver tissue were assessed according to Ludwig's criteria. RESULTS: In patients with chronic hepatitis C a clear relationship was found between CYP2E1 expression in the liver and the progression of hepatic disease (both lobular inflammation and fibrosis indices), and observed variations were consistent with the preferential distribution of CYP2E1 in the lobular zone. No effect of the liver disease was, however, found at the PBL level. A statistically significant increase in mean CYP2E1 expression level was observed in the lymphocytes from poorly controlled IDD subjects compared to controls. CONCLUSIONS: Taken together, our data indicate that the measurement of CYP2E1 expression in PBLs is not useful in liver diseases. However, in a systemic condition (IDD) this measurement can be proposed for monitoring the CYP2E1 induction in a relatively noninvasive manner. This tool should therefore be further validated in clinical field or experimental studies for CYP2E1 phenotyping purposes. PMID- 12743670 TI - Quinine 3-hydroxylation as a biomarker reaction for the activity of CYP3A4 in man. AB - OBJECTIVE: To investigate the usefulness of the 3-hydroxylation of quinine as a biomarker reaction for the activity of CYP3A4 in man and to study the interindividual variation in the metabolic ratio (MR), i.e. quinine/3 hydroxyquinine. METHODS: Data from a previous study (A) was used for determination of the MR of quinine in plasma and urine at different time points. In study B, 24 healthy Swedish subjects received 250 mg quinine hydrochloride first alone and later together with four other CYP probe drugs [losartan (CYP2C9), omeprazole (CYP2C19), debrisoquine (CYP2D6) and caffeine (CYP1A2)] administered on the same day. Plasma and urine samples were collected before quinine intake and 16 h thereafter and analysed for quinine and 3-hydroxyquinine using high-performance liquid chromatography. Plasma and/or urine were collected for the other probes at different time points. MRs of all the probes were determined and correlations to quinine MR were studied. RESULTS: In study A, the MR in plasma was stable over 96 h. The ratio increased from 5.8 to 12.2 (P=0.006) during co-administration with ketoconazole, whereas no significant difference (P=0.76) was observed during co-administration with fluvoxamine (from 5.8 to 6.0). In study B, there was no significant difference (P=0.36) between the mean MRs when quinine was given alone (4.7) or together with the four other drugs (4.5). There was a significant correlation between the MR of quinine and omeprazole sulphone formation (r=0.52, P<0.01), but not to the MRs of the other probes. There was a fivefold interindividual variability in the MR. CONCLUSIONS: The MR of quinine in plasma or urine may serve as a stable measure of the activity of CYP3A4 in man. These results together with in vitro data show that quinine is also a specific CYP3A4 probe. PMID- 12743673 TI - Polymorphic drug metabolism (CYP2D6) and utilisation of psychotropic drugs in hospitalised psychiatric patients: a retrospective study. AB - AIM: The aim of the current retrospective study was to assess the influence of polymorphic drug metabolism as assessed by genotyping, on the on the utilisation of psychotropic drugs in hospitalised psychiatric patients. The utilisation of psychotropic drugs was assessed using pharmacy records with emphasis on the number of prescriptions and prescriptions for possible side effects. METHODS: CYP2D6 genotype was assessed in 241 psychiatric patients by investigation for the five most common allelic variants ( CYP2D6*3, *4, *6, *7, *8) and the presence of gene duplication using allele-specific polymerase chain reaction. Data concerning the pharmacotherapy of the patients were retrieved from the pharmacy information system. Data was analysed on differences observed in pharmacy records concerning the different metabolic classes: ultra rapid metabolisers (UMs), extensive metabolisers (EMs) and poor metabolisers (PMs). RESULTS: For CYP2D6, 2.5% was UM (95% CI: 0.5-4.5%, n=6) and 8.3% was PM (95% CI: 4.8-11.8%, n=20). Drugs metabolised by CYP2D6 were less frequently prescribed in PMs than EMs (21.1% vs 33.6%, P=0.023). The average duration of prescriptions was significantly lower in PMs than EMs (54 days vs 106 days, P=0.010). Between UMs and EMs, no significant differences were found, although a similar tendency was observed. With regard to dose, no consistent differences were observed between the CYP2D6 genotype classes. Drugs against Parkinsonian-like side effects were given twice as frequently in PMs as EMs (6.9% vs 3.4%, P=0.045). CONCLUSIONS: Patients with impaired CYP2D6 metabolism received fewer CYP2D6 drugs. PMs were more prone to Parkinsonian-like side effects as evidenced by more prescriptions for drugs combating these side effects. Dose titrations were not often used to compensate for genetic polymorphisms. Pharmacy records might be a useful tool to detect differences related to polymorphic metabolism. PMID- 12743674 TI - Efficacy, safety and cost of new drugs acting on the central nervous system. AB - OBJECTIVE: To examine the degree of innovation of the products with indications for CNS diseases approved for the European market through the centralized procedure. METHODS: This paper examines the documentation available on nine products approved by the European Medicines Evaluation Agency in its first years of activity. RESULTS: The Committee for Proprietary Medicinal Products approved only five products by consensus (levacetylmethadol, levetiracetam, olanzapine, pramipexole, riluzole). Four were approved by majority (entacapone, memantine, rivastigmine, zaleplon). One product received a negative opinion, and five had the application withdrawn before reaching the Committee decision. CONCLUSIONS: An analysis of the efficacy and safety profile of these products indicates that few minor therapeutic advances have been achieved in this area. Most approved products cover needs already met, at higher cost, without substantial improvement. PMID- 12743672 TI - Pharmacokinetics and pharmacodynamics of ximelagatran, a novel oral direct thrombin inhibitor, in young healthy male subjects. AB - OBJECTIVES: Ximelagatran is a novel, oral direct thrombin inhibitor designed to overcome the low and variable oral absorption of melagatran, its active form. The pharmacokinetics and pharmacodynamics of ximelagatran following single and repeated oral administration were investigated. The primary objectives were to determine the dose linearity and reproducibility of melagatran exposure and the influence of food intake. METHODS: Two open-label studies were performed in healthy male subjects. Study I was a dose-escalation study, in which subjects received single oral doses of ximelagatran (1-98 mg). Study II was a randomised, two-way crossover study consisting of two 5-day treatment periods, in which subjects received a 20-mg oral dose of ximelagatran twice daily, either before breakfast and with dinner, or with breakfast and after dinner. RESULTS: Ximelagatran was rapidly absorbed and converted to melagatran, which was the predominant compound in plasma. The mean (+/- standard deviation) bioavailability of melagatran was 22.2+/-4.3% and 17.4+/-2.8% after single and repeated dosings, respectively. The maximum plasma concentration of melagatran and the area under the melagatran plasma concentration-time curve (AUC) increased linearly with dose. Inter- and intra-subject variability in melagatran AUC was 8% and 12%, respectively, with no relevant food- or time dependence. Anticoagulation, assessed as activated partial thromboplastin time, was correlated with melagatran plasma concentration. There was virtually no increase in capillary bleeding time over the dose range studied, and ximelagatran was well tolerated. CONCLUSION: After oral administration of ximelagatran to healthy male subjects, the pharmacokinetic and pharmacodynamic profile of melagatran is predictable and reproducible. PMID- 12743675 TI - Proprioceptive reflexes in patients with reflex sympathetic dystrophy. AB - Reflex sympathetic dystrophy (RSD) is a syndrome that frequently follows an injury and is characterized by sensory, autonomic and motor features of the affected extremities. One of the more common motor features of RSD is tonic dystonia, which is caused by impairment of inhibitory interneuronal spinal circuits. In this study the circuits that modulate the gain of proprioceptive reflexes of the shoulder musculature are quantitatively assessed in 19 RSD patients, 9 of whom presented with dystonia. The proprioceptive reflexes are quantified by applying two types of force disturbances: (1) disturbances with a fixed low frequency and a variable bandwidth and (2) disturbances with a small bandwidth around a prescribed centre frequency. Compared to controls, patients have lower reflex gains for velocity feedback in response to the disturbances around a prescribed centre frequency. Additionally, patients with dystonia lack the ability to generate negative reflex gains for position feedback, for these same disturbances. Proprioceptive reflexes to the disturbances with a fixed low frequency and variable bandwidth present no difference between patients and controls. Although dystonia in the RSD patients was limited to the distal musculature, the results suggest involvement of interneuronal circuits that mediate postsynaptic inhibition of the motoneurons of the proximal musculature. PMID- 12743676 TI - Long-loop reflex from arm afferents to remote muscles in normal man. AB - The present study was designed to examine the effects of median nerve stimulation on motoneurones of remote muscles in healthy subjects using H-reflex, averaged EMG and PSTH methods. Stimulation of the median nerve induced facilitation of soleus H-reflex from about 50 ms and it reached a peak at about 100 ms of conditioning-test interval. Afferents that induced the facilitation consisted of at least two types of fibres, the high-threshold cutaneous fibres and the low threshold fibres. When the effects were examined by the averaged surface EMG and PSTH, no facilitation but rather inhibition or inhibition-facilitation was induced in all tested muscles except for the upper limb muscles on the stimulated side. The inhibition latency was shortest in masseter muscle and longest in leg muscles, while values for the contralateral upper limb muscles were in the middle, indicating that the onset of inhibition was delayed from rostral to caudal muscles. Inputs from the median nerve converged to inhibitory interneurones, which mediate the masseter inhibitory reflex. Our findings suggested that inputs from the median nerve initially ascend to the brain, at least to the brainstem, and then descend to the spinal cord. Therefore, inhibition induced by median nerve stimulation was not considered as an interlimb reflex mediated by a propriospinal pathway, but long-loop reflex, at least via the pons. The discrepancy between the results of reflex and motor units suggests that facilitation of soleus H-reflex following median nerve stimulation was mainly due to reduced presynaptic inhibition. PMID- 12743677 TI - Spatial and temporal visual properties of single neurons in the feline anterior ectosylvian visual area. AB - The spatial and temporal visual sensitivity to drifting sinusoidal gratings was studied in 75 neurons of the feline anterior ectosylvian visual area (AEV). Extracellular single-unit recordings were performed in halothane-anesthetized (0.6%), immobilized, artificially ventilated cats. Most cells were strongly sensitive to the direction of drifting gratings. The mean value of the direction tuning widths was approximately 90 deg. Most of the cells (69 of the 75 cases) displayed rather narrowly tuned band-pass characteristics in the low spatial frequency range, with a mean optimal spatial frequency of 0.2 cycles/degree (c/deg). The mean spatial bandwidth was 1.4 octaves. The remainder of the units was low-pass tuned. A majority of the units responded optimally to high temporal frequencies (mean 6.3 Hz), although some cells did exhibit preferences for every examined temporal frequency between 0.6 Hz and 10.8 Hz. The temporal frequency tuning functions mostly revealed a band-pass character with a mean temporal bandwidth of 1.1 octaves. Our results demonstrate that the neurons along the anterior ectosylvian sulcus display particular spatial and temporal characteristics. The AEV neurons, with their preference for low spatial frequencies and with their fine spatial and temporal tuning properties, seem to be candidates for special tasks in motion perception. PMID- 12743678 TI - The effects of 5-hydroxytryptamine 5-HT2 receptor antagonists on nerve conduction velocity and endoneurial perfusion in diabetic rats. AB - Reduced peripheral nerve perfusion participates in the aetiology of diabetic neuropathy. 5-Hydroxtryptamine causes vasa nervorum vasoconstriction and platelet aggregation, which are enhanced by diabetes. To assess whether these mechanisms could contribute to neuropathy, the effects of 5-hydroxytryptamine 5-HT2 receptor antagonist treatment were examined in streptozotocin-induced diabetic rats. One study determined the dose-response relationship for AT1015 (N-[2-[4-(5H dibenzo[a,d]cyclohepten-5-ylidene)piperidino]ethyl]-1-formyl-4 piperidinecarboxamide monohydrochloride monohydrate). Two weeks AT1015 treatment after 6 weeks of diabetes dose-dependently corrected 19.7%, 54.1%, and 15.7% deficits in sciatic nerve motor conduction velocity and blood flow, and saphenous nerve sensory conduction: ED50 values were 0.52, 0.74 and 0.15 mg/kg(-1)/day(-1), respectively. In a second study, high-dose AT1015 (3 mg/kg(-1)/day(-1)) actions were compared with those of the 5HT2 receptor antagonists, ritanserin (10 mg/kg( 1)/day(-1)) and sarpogrelate (100 mg/kg(-1)/day(-1)), and the anti-platelet phosphodiesterase III inhibitor, cilostazol (100 mg/kg(-1)/day(-1)). Two weeks treatment with these drugs produced a marked correction (82.6-99.7%) of a 19.8% sciatic motor conduction deficit in diabetic rats. Similarly, 44.7% and 14.9% reductions in sciatic endoneurial blood flow and saphenous sensory conduction velocity were completely reversed. Thus, 5-HT2 receptor antagonists had marked beneficial effects in experimental diabetic neuropathy, and AT1015 appears suitable for further evaluation in clinical trials. PMID- 12743680 TI - The anabolic pyruvate oxidoreductase from Methanococcus maripaludis. AB - In autotrophic methanogens, pyruvate oxidoreductase (POR) plays a key role in the assimilation of CO(2) and the biosynthesis of organic carbon. This enzyme has been purified to homogeneity, and the genes from Methanococcus maripaludis were sequenced. The purified POR contained five polypeptides with molecular masses of 47, 33, 25, 21.5 and 13 kDa. The N-terminal sequences of four of the polypeptides had high similarity to the subunits commonly associated with this enzyme from other archaea. However, the 21.5-kDa polypeptide had not been previously observed in PORs. Nucleotide sequencing of the gene cluster encoding the POR revealed six open reading frames ( porABCDEF). The genes porABCD corresponded to the subunits previously identified in PORs. On the basis of the N-terminal amino acid sequence, porE encoded the 21.5-kDa polypeptide and contained a high cysteinyl residue content and a motif indicative of a [Fe-S] cluster. porF also had a high sequence similarity to porE, a high cysteinyl residue content, and two [Fe-S] cluster motifs. Homologs to porE were also present in the genomic sequences of the autotrophic methanogens Methanocaldococcus jannaschii and Methanothermobacter thermautotrophicus. Based upon these results, it is proposed that PorE and PorF are components of a specialized system required to transfer low-potential electrons for pyruvate biosynthesis. Some biochemical properties of the purified methanococcal POR were also determined. This unstable enzyme was very sensitive to O(2 )and demonstrated high activity with pyruvate, oxaloacetate, and alpha ketobutyrate. Methyl viologen, rubredoxin, FMN, and FAD were readily reduced. Activity was also observed with spinach and clostridial ferredoxins and cytochrome c. Coenzyme F(420) was not an electron acceptor for the purified enzyme. PMID- 12743679 TI - Repeated administration of the novel antipsychotic olanzapine does not modulate NMDA-sensitive glutamate and 5HT2 serotonin receptors in rats. AB - Antipsychotic drugs reportedly show a common property in facilitating glutamatergic transmission in rat cerebral cortex. Since the binding of the radiolabelled channel blocker [3H]-MK801 is generally considered an affordable index of N-methyl-d-aspartate (NMDA)-sensitive glutamate receptor activation, we examined the effects of clinically effective treatment (3 weeks, daily administration) of the atypical antipsychotic drug olanzapine (32 micromol/kg/5 ml) on the specific binding of [3H]-MK801 specific binding and on the strychnine insensitive glycine sites (glycine B) in synaptic plasma membranes (SPM) prepared from the medial prefrontal cortex (mPFC) of rats sacrificed after different (24, 60, 120 h) washout periods. We studied also the effects of repeated olanzapine administration on [3H]-ketanserin binding to 5HT2A receptors to verify whether, consistent with previously reported paradoxical effects of repeated administration of 5HT2A antagonists, this drug decreases 5HT2A receptor density without changing the apparent affinity. Neither single nor repeated olanzapine administration changed the kinetic characteristics of [3H]-MK801 or [3H]-glycine specific binding. When rats were sacrificed 120 h after the last olanzapine administration, both single or repeated treatment had failed to change the kinetic characteristics of [3H]-ketanserin binding, while the apparent affinity of 5HT2A receptors was increased in animals sacrificed at shorter (60 h) washout periods. Owing to the long half-life of olanzapine (24 h), and since the drug concentrations in mPFC of rats sacrificed 60 h after a single olanzapine administration (about 50 nM) are high enough to induce changes in 5HT2A receptor affinity, it is concluded that this modification, probably unrelated to the therapeutic efficacy, could be due to some drug still present in the brain at the time of the sacrifice. PMID- 12743681 TI - Medial meniscus interposition in proximal tibial epiphyseal fracture: a case report and review of the literature. AB - Proximal tibial fractures are rare in children, and most are treated conservatively. We report a case of a minimally displaced proximal tibial epiphyseal injury (Salter-Harris type III) in an 11-year-old child. We initially thought to treat this fracture conservatively, but the presence of massive haemarthrosis and considerable pain made us choose arthroscopy under general anaesthetic. During examination under anesthesia no instability could be detected, but the knee was locked in 5 degrees of flexion. On screening the fracture was not reducible anatomically. At arthroscopy the medial meniscus was not visualised as it was displaced into the fracture site. A small medial arthrotomy showed the meniscus trapped in the fracture site without any avulsion in the anterior or posterior horns. The fracture easily reduced once the meniscus was levered out. The importance of this injury is the fact that it could have been easily missed and would have led to serious functional deficit of the knee joint. We present radiographs and intra-operative pictures of this interesting injury. PMID- 12743682 TI - Risk factors of nosocomial catheter-associated urinary tract infection in a polyvalent intensive care unit. AB - OBJECTIVE: To determine the risk factors for catheter-associated urinary tract infection in a polyvalent intensive care unit (ICU). DESIGN: Prospective cohort study. SETTING: Sixteen-bed polyvalent ICU in a French university hospital. INTERVENTIONS: Prospective patient surveillance of patients included in two successive studies of two urine drainage systems. MAIN OUTCOME MEASURES: Bacteriuria occurrence in 553 ICU patients requiring a bladder catheter for longer than 48 h. The following variables were analyzed as possible risk factors: age, sex, severity score at admission, diagnosis on admission, duration of bladder catheterization, length of ICU stay, prior exposure to antibiotics, and system of urine drainage. RESULTS: The frequency of catheter-associated bacteriuria was 9.6%. From the multivariate analysis, five independent risk factors were determined: female sex, length of ICU stay, use of an antimicrobial therapy, severity score at admission, and duration of catheterization. CONCLUSION: In our study, the drainage system did not influence the occurrence of bacteriuria. To decrease the rate of catheter-associated bacteriuria in polyvalent ICU patients, removal of the bladder catheter must be performed as soon as possible. PMID- 12743683 TI - Schistosoma mansoni and Staphylococcus aureus bacteremia: a deadly association. PMID- 12743684 TI - [Rehabilitation after flexor tendon surgery]. AB - During the last 20 years there have been significant innovations in injury repair and aftercare for patients who sustain zone 2 flexor injuries. Based on improvements in our understanding of the mechanism of repair, new differentiated concepts could be developed. Active extension, passive flexion as introduced by Kleinert is still the standard in flexor tendon surgery. New stronger suture techniques allow immediate active flexion. The purpose of this article is to review the current concepts of flexor tendon and pulley repair as well as reconstruction of these structures and the rehabilitation programs for tenolysis. PMID- 12743685 TI - [Rehabilitation of extensor tendon injuries]. AB - The extensor mechanism of the hand consists of different tendons, originating from extrinsic and intrinsic muscles of the hand and forearm. Because of their course near to skin and bone separated only by a thin layer of paratenon, they are very sensitive to injuries and adhesions. Treatment protocols have to be adapted according to the different zones. This paper explains the basic principles of rehabilitation in extensor tendon injuries. PMID- 12743686 TI - [Rehabilitation after injuries to the metacarpal and finger joints]. AB - Finger joint injuries are the most frequent injuries of the hand. Besides operative and conservative therapy of these injuries, it is even more important to ensure a correct and thorough follow-up treatment. The aim of good rehabilitation is a painless, stabilized joint. Based on the therapeutic guidelines of the department of orthopedics, traumatology, and hand surgery of the Free University Berlin, the rehabilitation of these injuries is discussed. PMID- 12743687 TI - [Aftercare following distal forearm fractures and carpal trauma]. AB - Rehabilitation following operative or conservative treatment of fractures of the distal forearm and carpal trauma takes effect through adequate immobilization and ought to be maximized by a differentiated application of every means of physiotherapy, ergotherapy, and physical medicine available. Rehabilitation of the fractured distal forearm and carpal trauma are presented based on the therapeutic guidelines as applied by the Department of Hand Surgery, Plastic and Microsurgery of the Berufsgenossenschaftliches Unfallkrankenhaus Hamburg. PMID- 12743688 TI - [Rehabilitation after replantation]. AB - Limb replantation represents a particular surgical challenge. Rehabilitation and functional integration of the patient into everyday life has proven to be as equally important as the operation itself. This requires intensive and long-term cooperation between surgeon, therapist, and patient since replanted limbs without restoration of function are of no use. Besides physiotherapy, ergotherapy is highly important for this since it helps to reactivate daily activities.Also, tactile gnosis and protecting nervous sensibility and motor function can be improved by ergotherapy. With the help of special devices, adjustments,and psychological care, ergotherapy also supports the patient's reintegration into his former social environment. PMID- 12743689 TI - [Dupuytren's contracture: aftercare and long-term results]. AB - In addition to a surgical technique tailored to the disease stage, postoperative aftercare is important for favorable long-term results. An appropriate bandaging technique prevents formation of a postoperative hematoma,which promotes scar formation or development of relapsed Dupuytren's disease. PMID- 12743690 TI - [Nonoperative treatment of ischemic contractures of forearm and hand]. AB - The acute compartment syndrome of the forearm and hand leads to severe muscle necrosis and nerve damage if the diagnosis is not recognized. The resulting ischemic contractures and paralyses require a distinctive regime of therapy. Therefore,physiotherapy and occupational therapy are of main importance. In mild cases exclusively nonoperative treatment is possible. The goal is to exercise the function of the remaining muscles, mobilize the joints, and stretch muscle scars. Severe forms need surgery. To reach the best starting point for the operation, extensive preoperative active and passive exercises as well as splint therapy are required. Postoperatively, this regime has to be continued to retain a favorable outcome. The treatment approach is demonstrated with a case report. PMID- 12743691 TI - [The spastic hand. Postoperative rehabilitation]. AB - Postoperative rehabilitation starts with preoperative evaluation of the patient. The outcome of rehabilitation measures is strongly dependent on the surgeon's ability to define a realistic rehabilitation aim, i.e., a goal the patient can achieve with a high degree of probability. Therefore, knowledge of the patient's individual basic situation and status as well as of the autonomic development of the different categories of cerebral palsy is of great importance. The operative procedures used are well established in cerebral palsy surgery. They are accepted as standard procedures. The decisive question is to whom they are offered. The same conditions pertain to the postoperative rehabilitation period. During that time a combination of functional bracing and physiotherapy plus occupational therapy is offered after primary cast immobilization. This period is finished when the specific aim has been achieved, i.e., integration of the gained function into everyday life. PMID- 12743692 TI - [The artificial hand. An overview of hand prostheses]. AB - In spite of constant demands made by patients for optimal reconstruction of the hand in cases of injury or amputation, prosthetic devices allow only basic functions of the hand. The most important function, i.e., to connect sensitive tactile abilities of the hand with the eye and central nervous system, has not yet been achieved and will not be feasible in the near future. There have been numerous attempts to reconstruct single hand functions such as subtle finger control in all known directions etc. Until now none of these attempts have led to any serial production of a functional "tool" because the interface between prosthesis and CNS has not been sufficiently resolved. Minute finger control requires considerable mechanical engineering that includes an efficient motor drive and an adequate power supply,which renders possible prototypes for hand prostheses heavy and complicated in everyday performance. This survey offers a short and comprehensive introduction to high-performance but simple "down-to earth" orthotic and prosthetic devices for the replacement of lost forearm,hand, and finger function. PMID- 12743694 TI - [Complex regional pain syndrome type I (CRPS I). Pathophysiology, diagnostics, and therapy]. AB - Complex regional pain syndrome type I (CRPS type I)--formerly termed Sudeck's atrophy or reflex sympathetic dystrophy (RSD)--causes chronic, poorly controllable pain, autonomic, sensorimotor disorders,and serious trophic alterations in the later stages. It develops in the distal extremities mostly after minimal trauma or surgical intervention and rarely spontaneously. The severity of symptoms is disproportionate to the causative event. The latest scientific findings show that the previously called reflex sympathetic dystrophy (RSD), which was supposed to be a result of a hyperreactive autonomic nervous system,is a very complex syndrome that occurs on different integration levels of the nervous system. Sympathetically maintained pain (SMP) may be facultatively characteristic, but is not to be misunderstood as an underlying mechanism. A neurogenic inflammation reaction has recently been discussed, just as had been postulated by Paul Sudeck long before. That was the reason why the International Association for the Study of Pain (ISAP) introduced the more descriptive term "complex regional pain syndrome" (CRPS) type I in 1994. Due to the complexity of the process necessitating qualified knowledge, it is important to immediately refer patients to a specialized pain OPD or clinic. The diagnosis of CRPS type I is based upon a carefully taken case history and a clinical examination by an experienced practitioner. Imaging diagnostic tools and laboratory findings are of no or only low predicative value. The question of whether SMP exists after diagnosing CRPS type I is eminent for therapy planning. Therefore, diagnostic regional anesthetics are still important in spite of their uncertain prognostic relevance. Physical therapy, occupational therapy, medical treatment, and psychotherapy play an important role in the primary treatment of CRPS type I as noninvasive procedures. Despite heavy criticism, invasive sympathetic block, subsequent to adequate diagnostics, is an important part of the therapeutic concept. A multimodal therapeutic concept, which includes all available possibilities, is absolutely necessary to avoid grave permanent disabilities caused by insufficient or failed therapy. Nevertheless, already established as well as new treatment modalities have to be critically observed by further randomized, prospective control trials. PMID- 12743693 TI - [Treatment of hand surgery patients in chronic pain]. AB - The treatment of hand surgery patients suffering from chronic pain requires an interdisciplinary procedure. An imbalance between nociception and antinociception can be seen as a reason for the chronification of pain. The complexity of the problem of chronic pain patients is marked by a wide variety of symptoms. Full diagnostic assessment to detect or to exclude other organic diseases is required. Rehabilitation to improve function and reduce pain intensity has priority. Our therapeutic strategy consists of intensive physiotherapy and analgesic drugs as well as a series of blockades with buprenorphine (Temgesic) of the ganglion stellatum (GLOA). This specific therapy achieved an improvement of the function of the upper extremity and a reduction of pain intensity. The majority of patients were satisfied with the outcome. PMID- 12743695 TI - [Tossy III injuries of the acromioclavicular joint. In what circumstances is surgery still justified? Personal results and literature review]. AB - In Germany AC-joint-dislocations type Tossy III are treated in most of the cases operatively. Over two times of period we treated AC-joint-dislocations type Tossy III with biodegradable PDS-cords. 54 patients were operated between 1989 and 1997 and followed up after 39 months. 87.5% of patients are satisfied with results and have 10.2 points at Taftscore. In this period we couldn't differentiated the results by Rockwood classification. 12 patients with Rockwood V were operated between 1998 and 2002 and followed up after 14 months. Excellent and good results were seen in 92% of cases. At Taftscore we seen 10.7 points. 3 patients with Rockwood III were treated conservative with 10.3 points and 3 patients were operated with 10.7 points at Taftscore. AC-joint dislocations should be classified to Rockwood. No differences were seen between operation and conservative treatment in Rockwood III in literature. We recommend operation with PDS cords with good results in Rockwood V. PMID- 12743696 TI - [Aplasia of the posterior tibial artery in a child with idiopathic clubfoot]. AB - A case of a completely absent posterior tibial artery discovered during a procedure for congenital clubfoot correction in a 6 month old male infant is reported. Anomalies of the anterior tibial artery associated with the clubfoot deformity are common, the absence of the posterior tibial artery is, however, very rare. PMID- 12743697 TI - NF-kappaB in epiretinal membranes after human diabetic retinopathy. AB - AIMS/HYPOTHESIS: Formation of epiretinal membranes (ERMs) in the posterior fundus results in progressive deterioration of vision. ERMs have been associated with numerous clinical conditions including proliferative diabetic retinopathy (PDR), but its pathogenic mechanisms are still unknown. This study was conducted to examine whether or not nuclear factor kappa B (NF-kappaB), a transcription factor that can be activated by various pathological conditions, is involved in the formation of ERMs after PDR. METHODS: ERM samples were obtained by vitrectomy from 22 cases with PDR aged 56+/-11 years with 18+/-10 years of diabetes and 15 cases with idiopathic ERM. They were processed for reverse transcription polymerase chain reaction (RT-PCR) analysis. In addition, 5 ERM samples from PDR patients aged 51+/-16 years with 15+/-6 years of diabetes were processed for immunohistochemical analysis. RESULTS: NF-kappaB mRNA expression levels were higher (20 out of 22 cases vs. 9 out of 15 subjects in idiopathic ERM, p<0.05) in PDR subjects. Immunohistochemical analysis showed NF-kappaB protein expression in all the 5 ERMs derived from PDR patients, and that region was partially double labelled with interleukin-8 (IL-8) and von Willebrand factor (vWF). CONCLUSIONS/INTERPRETATION: These results suggest a possibility that NF-kappaB is involved in the formation of ERMs after PDR, especially for the development of vascular endothelial cell component. PMID- 12743699 TI - The number of lactotrophs is reduced in the anterior pituitary of streptozotocin induced diabetic rats. AB - AIMS/HYPOTHESIS: Prolactin secretion is often reduced in insulin dependent diabetes mellitus, but little is known about the mechanism involved. Since changes in the hormonal environment modulate cell proliferation, death and cellular makeup of the anterior pituitary, we have analysed whether the number of lactotrophs is reduced in diabetic rats. METHODS: Streptozotocin induced diabetic rats were maintained hyperglycaemic for 2 months. Pituitary prolactin, growth hormone, Bcl-2, Bax and PCNA concentrations were analysed by western blot analysis. In situ hybridisation was used for quantification of prolactin and growth hormone mRNA containing cells. Cell death was detected by TUNEL labelling, alone and in combination with immunocytochemistry for prolactin or growth hormone. RESULTS: Diabetic rats had fewer lactotrophs ( p<0.01). This was coincident with a decrease in overall protein and prolactin content. An increase in pituitary cell death was found and some of the TUNEL labelling co-localised with prolactin immunostaining. No change in the concentration of Bcl-2 or Bax, proteins implicated in apoptosis, was detected. PCNA content was higher in the pituitaries of diabetic rats, suggesting increased proliferation. CONCLUSION/INTERPRETATION: Anterior pituitary cell turnover is affected in poorly controlled diabetes mellitus. A decrease in the number of lactotrophs, as a result of increased cell death, could underlie, at least in part, the reduction in prolactin secretion observed in diabetic animals. PMID- 12743698 TI - Kallikrein-binding protein inhibits retinal neovascularization and decreases vascular leakage. AB - AIMS/HYPOTHESIS: Kallikrein-binding protein (KBP) is a serine proteinase inhibitor (serpin). It specifically binds to tissue kallikrein and inhibits kallikrein activity. Our study was designed to test its effects on retinal neovascularization and vascular permeability. METHODS: Endothelial cell proliferation was determined by [(3)H] thymidine incorporation assay and apoptosis quantified by Annexin V staining and flow cytometry. Effect on retinal neovascularization was determined by fluorescein angiography and count of pre retinal vascular cells in an oxygen-induced retinopathy (OIR) model. Vascular permeability was assayed by the Evans blue method. Vascular endothelial growth factor (VEGF) was measured by Western blot analysis and ELISA. RESULTS: Kallikrein-binding protein specifically inhibited proliferation and induced apoptosis in retinal capillary endothelial cells. Intravitreal injection of KBP inhibited retinal neovascularization in an OIR model. Moreover, KBP decreased vascular leakage in the retina, iris and choroid in rats with OIR. Blockade of kinin receptors by specific antagonists showed significantly weaker inhibition of endothelial cells, when compared to that of KBP, suggesting that the anti angiogenic activity of KBP is not through inhibiting kallikrein activity or kinin production. KBP competed with (125)I-VEGF for binding to endothelial cells and down-regulated VEGF production in endothelial cells and in the retina of the OIR rat model. CONCLUSION/INTERPRETATION: Kallikrein-binding protein is a multi functional serpin, and its vascular activities are independent of its interactions with the kallikrein-kinin system. Inhibition of VEGF binding to its receptors and down-regulation of VEGF expression could represent a mechanism for the vascular activities of KBP. PMID- 12743700 TI - The polymorphism Gly574Ser in the transcription factor HNF-1alpha is not a marker of adult-onset ketosis-prone atypical diabetes in Afro-Caribbean patients. PMID- 12743701 TI - Lipoprotein subclass measurements by nuclear magnetic resonance spectroscopy improve the prediction of coronary artery disease in Type 1 diabetes. A prospective report from the Pittsburgh Epidemiology of Diabetes Complications Study. AB - AIM/HYPOTHESIS: To examine whether nuclear magnetic resonance lipoprotein spectroscopy improves the prediction of coronary artery disease in patients with Type 1 diabetes, independently of conventional lipid and other risk factors. METHODS: A prospective nested case-control design of subjects with childhood onset Type 1 diabetes from the Pittsburgh Epidemiology of Diabetes Complications Study was used. 59 controls were age-, sex- and duration-matched to 59 incident cases of coronary artery disease (fatal or non-fatal myocardial infarction, angina, coronary stenosis >50%) occurring during 10 years of follow-up. Lipid mass and particle concentrations of VLDL, LDL, and HDL subclasses, grouped into three size categories (large, medium, and small), were assessed prior to event with nuclear magnetic resonance spectroscopy. RESULTS: Univariate analyses showed that both lipid mass and particle concentrations of all three VLDL subclasses, small LDL, medium LDL, and medium HDL were increased in CAD cases compared to controls, while large HDL was decreased. Mean LDL and HDL particle sizes were lower in cases. In multivariate models using conventional lipid and non-lipid risk factors, triglycerides and overt nephropathy were the strongest predictors of CAD. Nuclear magnetic resonance measures further improved the prediction, i.e. large HDL particle concentration (OR=0.43, p=0.030), medium HDL mass (OR=3.79, p=0.026) and total VLDL particle concentration (OR=2.33, p=0.033). CONCLUSION/INTERPRETATION: While these results underscore the importance of triglycerides and overt nephropathy in CAD risk in Type 1 diabetic patients, they also suggest that nuclear magnetic resonance lipoprotein spectroscopy could further refine its prediction and show novel findings concerning HDL subclasses. PMID- 12743703 TI - Thermoregulation and ventilation of termite mounds. AB - Some of the most sophisticated of all animal-built structures are the mounds of African termites of the subfamily Macrotermitinae, the fungus-growing termites. They have long been studied as fascinating textbook examples of thermoregulation or ventilation of animal buildings. However, little research has been designed to provide critical tests of these paradigms, derived from a very small number of original papers. Here I review results from recent studies on Macrotermes bellicosus that considered the interdependence of ambient temperature, thermoregulation, ventilation and mound architecture, and that question some of the fundamental paradigms of termite mounds. M. bellicosus achieves thermal homeostasis within the mound, but ambient temperature has an influence too. In colonies in comparably cool habitats, mound architecture is adapted to reduce the loss of metabolically produced heat to the environment. While this has no negative consequences in small colonies, it produces a trade-off with gas exchange in large colonies, resulting in suboptimally low nest temperatures and increased CO(2) concentrations. Along with the alteration in mound architecture, the gas exchange/ventilation mechanism also changes. While mounds in the thermally appropriate savannah have a very efficient circular ventilation during the day, the ventilation in the cooler forest is a less efficient upward movement of air, with gas exchange restricted by reduced surface exchange area. These results, together with other recent findings, question entrenched ideas such as the thermosiphon-ventilation mechanism or the assumption that mounds function to dissipate internally produced heat. Models trying to explain the proximate mechanisms of mound building, or building elements, are discussed. PMID- 12743704 TI - Jeholornis compared to Archaeopteryx, with a new understanding of the earliest avian evolution. AB - The recently reported Jeholornis represents the only known bird with a complete long skeletal tail except for Archaeopteryx. Two newly discovered specimens referable to Jeholornis provide some important new information about its anatomy. The tail of Jeholornis is much longer than that of Archaeopteryx and comprises a maximum of 27 caudal vertebrae compared with only 23 in Archaeopteryx. More interestingly, the tail feathers are shaped more like those of dromaeosaurs than those of Archaeopteryx. We conclude that the common ancestor of birds must have a more primitive tail than that in Archaeopteryx, confirming the side branch position of Archaeopteryx in the early avian evolution. The synsacrum is composed of six sacrals, representing a transitional stage between Archaeopteryx and more advanced birds. The scapula of Jeholornis has a dorso-laterally exposed glenoid facet, and the coracoid has a supracoracoid foramen. The presence of a pair of fenestrae in the sternum of Jeholornis has further implications for the air-sac system in early birds. PMID- 12743702 TI - Hox in hair growth and development. AB - The evolutionarily conserved Hox gene family of transcriptional regulators has originally been known for specifying positional identities along the longitudinal body axis of bilateral metazoans, including mouse and man. It is believed that subsequent to this archaic role, subsets of Hox genes have been co-opted for patterning functions in phylogenetically more recent structures, such as limbs and epithelial appendages. Among these, the hair follicle is of particular interest, as it is the only organ undergoing cyclical phases of regression and regeneration during the entire life span of an organism. Furthermore, the hair follicle is increasingly capturing the attention of developmental geneticists, as this abundantly available miniature organ mimics key aspects of embryonic patterning and, in addition, presents a model for studying organ renewal. The first Hox gene shown to play a universal role in hair follicle development is Hoxc13, as both Hoxc13-deficient and overexpressing mice exhibit severe hair growth and patterning defects. Differential gene expression analyses in the skin of these mutants, as well as in vitro DNA binding studies performed with potential targets for HOXC13 transcriptional regulation in human hair, identified genes encoding hair-specific keratins and keratin-associated proteins (KAPs) as major groups of presumptive Hoxc13 downstream effectors in the control of hair growth. The Hoxc13 mutant might thus serve as a paradigm for studying hair specific roles of Hoxc13 and other members of this gene family, whose distinct spatio-temporally restricted expression patterns during hair development and cycling suggest discrete functions in follicular patterning and hair cycle control. The main conclusion from a discussion of these potential roles vis-a-vis current expression data in mouse and man, and from the perspective of the results obtained with the Hoxc13 transgenic models, is that members of the Hox family are likely to fulfill essential roles of great functional diversity in hair that require complex transcriptional control mechanisms to ensure proper spatio temporal patterns of Hox gene expression at homeostatic levels. PMID- 12743705 TI - To see or not to see: does previewing a future opponent affect the contest behavior of green swordtail males ( Xiphophorus helleri)? AB - Animals assess the fighting ability of conspecifics either by engaging in aggressive interactions or observing contests between others. However, whether individuals assess physical prowess outside the context of aggressive interactions remains unknown. We examined whether male green swordtails ( Xiphophorus helleri) extract information about the fighting ability of solitary individuals via observation and whether acquiring such information elicits behavioral modifications. Contests preceded by mutual visual assessment were significantly shorter than fights where only one or neither of the two individuals was informed in advance. Focal animals initiated aggressive behavior more often against larger opponents only after previewing their adversary, indicating that swordtails can extract information about relative body size from watching solitary conspecifics. When a fighting disadvantage is perceived, observers adopt tactics that increase their probability of winning the contest. PMID- 12743706 TI - Phylogeny of cooperatively breeding cuckoos (Cuculidae, Crotophaginae) based on mitochondrial gene sequences. AB - The Crotophaginae is a subfamily of New World cuckoos comprising the monotypic genus Guira and three ani species ( Crotophaga). All exhibit a rare form of cooperative breeding known as plural female joint-nesting, whereby two or more females lay eggs in a single nest. I reconstructed the phylogeny of Crotophaginae using the mitochondrial genes cytochrome oxidase I, II, and III, ATPase 6 and 8, and cytochrome b. The subfamily was monophyletic, implying a single origin of cooperative breeding in New World cuckoos. Crotophaga was also monophyletic with Guira as its sister taxon. Within Crotophaga, the smooth-billed ( C. ani) and groove-billed ( C. sulcirostris) anis formed the internal clade with the greater ani ( C. major) basal to this pair. This phylogeny is consistent with differences in reproductive patterns and social organization exhibited by crotophagine cuckoos, and will serve as a framework for future study of the evolution of cooperative breeding in this subfamily. PMID- 12743707 TI - Plumage coloration and nutritional condition in the great tit Parus major: the roles of carotenoids and melanins differ. AB - The size and coloration of some body characters seem to influence mate choice in many species. Most animal colours are either structural or based on melanin or carotenoid pigments. It has recently been suggested that carotenoid-based or structural coloration may be a condition-dependent trait, whereas melanin-based coloration is not; a difference that may be highly relevant when studying the evolution of multiple mating preferences. We tested this hypothesis in the great tit ( Parus major). The size of the melanin breast band was not correlated to nutritional condition as estimated by the rate of tail growth (ptilochronology), controlling for locality, age, sex, year and season effects. However, the correlation was significant for the hue of yellow breast (carotenoid-based coloration), and the slopes of the regressions of the two pigments to growth bars differed significantly. These results suggest that the expression of the two traits may be regulated by different mechanisms. PMID- 12743708 TI - Food offering in jackdaws ( Corvus monedula). AB - Food sharing among unrelated same-sex individuals has received considerable interest from primatologists and evolutionary biologists because of its apparent altruistic nature and implications for the evolution of complex social cognition. In contrast to primates, food sharing in birds has received relatively little attention. Here we describe three types of food sharing in jackdaws, with the initiative for the transfer either with the receiver or the giver. The latter situation is of particular interest because the food transfer takes place through active giving. Compared to primates, jackdaws show high rates of food sharing. Finally we discuss the implications of food sharing in jackdaws, and in birds in general. PMID- 12743709 TI - Nuclear factor kappa B is activated in small intestinal mucosa of celiac patients. AB - NF-kappa B regulates inflammatory and immune response by increasing the expression of specific genes. In celiac disease proinflammatory cytokines, adhesion molecules, and enzymes whose gene expression is known to be regulated by NF-kappa B are involved. This study investigated the activation of NF-kappa B in inflamed mucosa from patients with untreated celiac disease. Biopsy specimens from control, untreated, and treated patients were subjected to molecular biology analysis. NF-kappa B activation was evaluated by electrophoretic mobility shift assay. NF-kappa B related subunit protein level, and inducible nitric oxide synthase and cyclo-oxygenase 2 protein expression was analyzed by western blot. Both NF-kappa B/DNA binding activity and p50/p65 nuclear levels were higher in biopsy specimens from untreated patients than in those from treated patients and controls. The degradation of I kappa B beta in the cytosol and the reappearance in the nucleus indicated a persistent NF-kappa B activation in celiac disease. NF kappa B activity was maintained in cultured biopsy specimens up to 6 h and decreased at 24 h, and then the addition of peptic-tryptic digest of gliadin caused the recovery of NF-kappa B activity at 6 h. NF-kappa B/DNA binding activity was correlated with inducible nitric oxide synthase and cyclo-oxygenase 2 protein expression. These results show for the first time that NF-kappa B is activated in the inflamed mucosa of celiac patients and suggest that it may represent a molecular target for the modulation of inflammatory response in celiac disease. PMID- 12743710 TI - High frequency of polymorphism Arg753Gln of the Toll-like receptor-2 gene detected by a novel allele-specific PCR. AB - The recently described family of Toll-like receptors (TLRs) plays a major role in innate immunity by mediating inflammatory reactions against a wide array of pathogens. TLR-2 is reported to interact with various bacterial partial structures including lipoproteins, peptidoglycan, and lipoteichoic acid. Two polymorphisms of the TLR-2 gene have recently been described: Arg753Gln, correlated with the incidence of sepsis in a white population, and Arg677Trp, correlated with the incidence of lepromatous leprosy in an Asian population. Both polymorphisms, when inserted into expression vectors encoding for human TLR-2, reduced stimulation of Chinese hamster ovary cells by synthetic lipopeptides. We furthermore developed a rapid and inexpensive method for the detection of both single nucleotide polymorphisms based on restriction fragment length polymorphism. While no individuals carrying the Arg677Trp SNP were identified in a large group of whites, 9.4% of the study population were found to be heterozygous for the Arg753Gln polymorphism. This ratio is significantly higher than previously reported, and therefore detection of this polymorphism among patients may yield important information for the assessment of risk profiles regarding susceptibility to bacterial infections. PMID- 12743711 TI - A walk though vertebrate and invertebrate protamines. AB - An updated comparative analysis of protamines and their corresponding genes is presented, including representative organisms from each of the vertebrate classes and one invertebrate (squid, Loligo opalescens). Special emphasis is placed on the implications for sperm chromatin organization and the evolutionary significance. The review is based on some of the most recent publications in the field and builds upon previously published reviews on this topic. PMID- 12743712 TI - Roles of transition nuclear proteins in spermiogenesis. AB - The transition nuclear proteins (TPs) constitute 90% of the chromatin basic proteins during the steps of spermiogenesis between histone removal and the deposition of the protamines. We first summarize the properties of the two major transition nuclear proteins, TP1 and TP2, and present concepts, based on their time of appearance in vivo and in vitro properties, regarding their roles. Distinct roles for the two TPs in histone displacement, sperm nuclear shaping, chromatin condensation, and maintenance of DNA integrity have been proposed. More definitive information on their roles in spermiogenesis has recently been obtained using mice with null mutations in the Tnp1 or Tnp2 genes for TP1 and TP2, respectively. In these mice, histone displacement and sperm nuclear shaping appear to progress quite normally. Spermatid nuclear condensation occurs, albeit in an abnormal fashion, and the mature sperm of the Tnp -null mutants are not as condensed as wild-type sperm. There is also evidence that sperm from these mutant mice contain an elevated level of DNA strand breaks. The mutant sperm showed several unexpected phenotypes, including a high incidence of configurational defects, such as heads bent back on midpieces, midpieces in hairpin configurations, coils, and clumps, other midpiece defects, reduced levels of proteolytic processing of protamine 2 during maturation, and reduced motility. The two TPs appear partly to compensate for each other as both Tnp1 - and Tnp2 null mice were able to produce offspring, and appear to have largely overlapping functions as the two mutants had similar phenotypes. PMID- 12743714 TI - Endoscopic CO2 laser-assisted surgery for cricopharyngeal dysfunction. AB - Endoscopic cricopharyngeal myotomy was performed on 29 patients with dysphagia from failed relaxation of the cricopharyngeal muscle. The patient outcome was retrospectively evaluated. The average age at the time of treatment was 62 years (range: 38-81 years), and the mean follow-up was 18 months (range:1-36 months). The procedure was the first to be performed in all except four of the patients. Preoperative and postoperative assessments included videofluoroscopic and flexible endoscopic evaluations of the swallow as well as patients' subjective ratings for dysphagia and aspiration. These investigations and self-assessments were rated from 0 (poor or abnormal) to 4 (good or normal). Surgical procedures were performed under general anesthesia. Using the diverticuloscope, the posterior portion of the cricopharyngeal muscle was exposed and CO2 laser sectioned. The wound was then covered with fibrin glue. Patients were parenterally fed for 72 h. Postoperative videofluoroscopy showed the absence of leakage, and all patients resumed oral intake on day 2. The median self-rating score improved from 1 to 4 for dysphagia and from 3 to 4 for aspiration. The outcome of the flexible endoscopic evaluation of swallow improved from 2 to 4 and videofluoroscopy improved from 2 to 4. No surgical complication occurred. Endoscopic CO2 laser-assisted surgery is an effective and safe alternative for the treatment of cricopharyngeal dysmotility. PMID- 12743715 TI - Histological findings of the femoral bone after cement removal in hip revision. An experimental study of cadaver femurs with two different cement removal procedures. AB - Cement removal in hip revision arthroplasty is often a time-consuming procedure, lengthy and tedious. Intraoperative bone damage is one of the more common complications. In the present study, the conventional cement removal method is compared with a new method by means of a histological study concerning potential negative effects to cortical or spongious bone. Histological studies on human cadaver femurs demonstrate no deleterious effects on the endosteal bone when cement was removed with this new device. The ballistically driven chiselling system (OrthoClast) is safe to the bone stock and shows no increased risk of bone damage over the conventional technique with mallet and chisel. PMID- 12743716 TI - A comparative clinical outcome evaluation of smooth (10-13 year results) versus rough surface finish (5-8 year results) in an otherwise identically designed cemented titanium alloy stem. AB - BACKGROUND: We retrospectively compared the influence of surface finish with respect to the long-term durability of a cemented titanium alloy stem of the same design but different surface finish. METHODS: From 1984 to 1994, the stem was made of a titanium-aluminum-vanadium alloy. A total of 201 patients with 220 smooth stems (mean follow-up 11 years and 4 months) out of 612 implanted between 1984 and 1987 and 319 patients with 343 rough stems (mean follow-up 5 years and 10 months) out of 812 implanted between 1991 and 1993 could be reviewed clinically and radiologically for comparison. The average age of the patients with the smooth stem was 58.1 years and of the patients with the rough stem, 62.2 years. In both groups, 35% of patients were male and 65% female. RESULTS: The mean Merle d'Aubigne hip score increased from 10.5 to 16.0 points in the smooth stem group and from 11.0 to 16.9 points in the rough stem group. The distribution of radiolucent lines, according to the zones of Gruen, was similar in both groups. The smooth stem required revision in 18 cases after a mean follow-up of 11 years and 4 months and the rough stem in 30 cases after a mean follow-up of 5 years and 10 months. The survival analysis (Kaplan-Meier) revealed 95.4% survival after 13 years for the smooth femoral component and 76.7% survival after 8 years for the rough femoral component. CONCLUSIONS: The implantation of titanium alloy stems with a rough surface finish cannot be recommended because of its high aseptic loosening rate. PMID- 12743713 TI - Chromatin remodeling by nuclear receptors. AB - The eukaryotic genome is structurally organized into nucleosomes to form chromatin, which regulates gene expression, in part, by controlling the accessibility of regulatory factors. When packaged as chromatin, many promoters are transcriptionally repressed, thus reducing the access of transcription factors to their binding sites. However, nuclear receptors (NRs) are a group of transcription factors that have the ability to access their binding sites in this repressive chromatin structure. Nuclear receptors are able to bind to their sites and recruit chromatin-remodeling proteins such as ATP-dependent chromatin remodeling complexes and histone-modifying enzymes, resulting in transcriptional activation. In this review, we present the role of NRs in recruiting these chromatin-modifying enzymes by means of an extensively studied model system, the glucocorticoid receptor-mediated transactivation of the mouse mammary tumor virus (MMTV) promoter. We use these findings as a template to begin to understand the effect of chromatin changes on gene expression during spermatogenesis. PMID- 12743718 TI - Encephaloduroarteriosynangiosis with bifrontal encephalogaleo(periosteal)synangiosis in the pediatric moyamoya disease: the surgical technique and its outcomes. AB - METHODS: To increase the blood flow of the anterior cerebral artery (ACA) and the middle cerebral artery (MCA) territories, we modified the "ribbon" procedure in combination with encephaloduroarteriosynangiosis (EDAS). This is referred to as "EDAS with bifrontal encephalogaleo(periosteal)synangiosis (EGS)." The surgical technique, clinical outcomes, complications, extent of revascularization, and changes in CBF in 67 pediatric MMD patients were retrospectively reviewed. RESULTS: The excellent and good clinical recovery rates were 57% and 31%. The rate for complete disappearance of TIA was 63%. All the bifrontal EGS made abundant collateral vessels in the ACA territory. When the EDAS with bifrontal EGS was performed in the first operation, collaterals of EGS sites developed more on the contralateral side of the EDAS. The arachnoid opening of the medial frontal lobe in the EGS site had no effect on the results. There was a positive correlation between the clinical outcome and the extent of angiographic revascularization. Improvements in the CBF and the reserve in ACA territory were observed in 57%. CONCLUSIONS: EDAS with bifrontal EGS resulted in excellent revascularization in both the MCA and ACA territories. The clinical and hemodynamic results were also excellent. This procedure may be an effective and safe surgical modality for the prevention of ischemia in the whole territory of the anterior circulation of the brain in pediatric MMD. PMID- 12743719 TI - Progressive quadriparesis in adolescent with stenosis of the cervical spine. AB - INTRODUCTION: Progressive quadriparesis in adolescents suffering from cervical stenosis is a very rare entity. Only three cases have been reported in the literature. CASE REPORT: We report our own case of progressive quadriparesis in a young patient suffering from cervical stenosis, the first to be documented with pre- and postoperative magnetic resonance imaging. PMID- 12743720 TI - Effect of nitric oxide on electrolyte transport across the porcine proximal colon. AB - The effect of nitric oxide (NO) on ion transport in the porcine proximal colon was investigated in slide-stripped epithelia mounted in Ussing chambers. The serosal addition of the NO-donors sodium nitroprusside (SNP, 0.5 mM) or S-nitroso N-acetylpenicillamine (SNAP, 0.5 mM) induced a steep increase of short-circuit current ( I(sc)). The stimulatory effect of SNP on I(sc) could not be blocked by piroxicam or tetrodotoxin. Potassium channel inhibitors (quinidine, tetraethylammonium or barium) added serosally reduced the SNP- or SNAP-induced increases of I(sc). In chloride-free solutions, the SNP-induced increase of I(sc) was smaller than in chloride-containing solutions. Cl(- )and Na(+) flux measurements demonstrated that SNP diminished Cl(-) and Na(+) net absorption. Pre treatment with barium was able to block the inhibitory effect of SNP on NaCl net absorption totally. NO effects on paracellular pathways were assessed by measuring flux rates of [(14)C]-D-mannitol. SNP did not change unidirectional D mannitol flux rates. In conclusion, NO inhibits NaCl net absorption in the proximal colon of pigs by acting directly on the enterocyte. The antiabsorptive (and/or prosecretory) effect of NO depends on a functional basolateral potassium conductance. PMID- 12743721 TI - Intestinal passive absorption of water-soluble compounds by sparrows: effect of molecular size and luminal nutrients. AB - We tested predictions that: (1) absorption of water-soluble probes decreases with increasing molecular size, consistent with movement through effective pores in epithelia, and (2) absorption of probes is enhanced when measured in the presence of luminal nutrients, as predicted for paracellular solvent drag. Probes (L arabinose, L-rhamnose, perseitol, lactulose; MW 150.1-342.3 Da) were gavaged in nonanesthetized House sparrows ( Passer domesticus), or injected into the pectoralis, and serially measured in plasma. Bioavailability was calculated as F=AUC by gavage/AUC by injection, where AUC is the area under the curve of plasma probe concentration vs. time. Consistent with predictions, F declined with probe size by 75% from the smallest to the largest probe, and absorption of probes increased by 40% in the presence of luminal glucose or food compared to a mannitol control. Absorption of water-soluble probes by sparrows is much higher than in humans, which is much higher than in rats. These differences seem mainly attributable to differences in paracellular solvent flux and less to differences in effective paracellular pore size. PMID- 12743722 TI - Changes in tissue fatty acid composition during the first month of growth of the king penguin chick. AB - The switch from yolk to food (myctophid fishes) as the nutrient source for the newly hatched chick of the king penguin ( Aptenodytes patagonicus) results in a profound change in the pattern of fatty acid provision. This is characterized by major increases in the proportionate intake of n-3 polyunsaturates (20:5n-3 and 22:6n-3) and long chain (C(20-24)) monounsaturates, accompanied by relatively lower levels of n-6 polyunsaturates (18:2n-6 and 20:4n-6). The effects of this change on the fatty acid composition of tissue lipids during the first month of growth, a period of tissue maturation leading to thermal emancipation, were determined. The composition of adipose tissue triacylglycerol responded rapidly to the switch in nutrient source, the proportion of long chain monounsaturates (mainly 20:1n-9 and 22:1n-11) increasing five-fold between hatch and emancipation while the relative levels of 20:5n-3 and 22:6n-3 also increased significantly, by 3- and 1.2-fold, respectively. At emancipation, the fatty acid profile of adipose tissue triacylglycerol was essentially identical to that of the diet. At hatch, the main polyunsaturates of muscle phospholipid were 20:4n-6, 20:5n-3, and 22:6n 3, respectively, forming (w/w of fatty acids) 13.2%, 5.0%, and 12.0%. By emancipation, 20:4n-6 had decreased to 4.8%, 20:5n-3 increased to 10.9%, and 22:6n-3 at 11.4% showed little change. The main polyunsaturate in brain phospholipid at hatch was 22:6n-3 (19.3%): this remained almost constant until day 15 but then increased significantly to 23.6% by emancipation. Significant but minor changes in the proportions of 20:4n-6 (from 5.2% at hatch to 3.5% at emancipation) and 20:5n-3 (from 3.0% to 3.9%) were also observed in brain phospholipid. The data do not allow us to completely distinguish changes that are solely diet driven from those which are a consequence of tissue differentiation. Nevertheless, it is evident that, whereas the fatty acid composition of adipose tissue responds faithfully to the change in nutrient source, the phospholipids of muscle and, especially, of brain are much more refractory to the effects of diet during this period of tissue maturation. PMID- 12743723 TI - Heat increment of feeding and thermal substitution in mallard ducks feeding voluntarily on grain. AB - The heat increment of feeding (HIF), including heat from digestion, assimilation, and nutrient interconversion, may substitute for thermogenesis and reduce thermoregulation costs. HIF and its substitution have been measured mainly in animals fed single large meals with high protein content, but many species such as some dabbling ducks (Anatini) feed more continuously in intermittent small meals with low protein content. We measured HIF in seven mallard ducks (Anas platyrhynchos) eating mixed grain (corn, wheat, milo) ad libitum while floating on water at 23 degrees C (thermoneutral) and 8 degrees C. HIF was calculated as the difference in oxygen consumption between fed and fasted birds, correcting for costs of behavior, heat storage (change in body temperature), and heating food. Substitution occurred if HIF was lower at 8 degrees C than at 23 degrees C. Food intake of mallards averaged 83% of that required for maintenance (zero energy balance) at 23 degrees C, and 68% of maintenance at 8 degrees C. Mean HIF (+/-1 SE) was 1.59+/-0.61 l O(2) at 23 degrees C and 1.48+/-0.68 l O(2) at 8 degrees C. These values were 4.9% and 3.9% of metabolizable energy intake, consistent with values expected for grain. HIF did not differ between temperatures (ANCOVA, birds as blocks, intake as covariate, P=0.51), indicating no measurable substitution at these intake levels in intermittent meals. For these large birds that feed on low protein foods in intermittent small meals, the ecological importance of HIF substitution appears negligible during periods when food intake is below that required for energy balance. PMID- 12743724 TI - Thermoregulatory physiology of the Crested Pigeon Ocyphaps lophotes and the Brush Bronzewing Phaps elegans. AB - The metabolic physiology of the Crested Pigeon (Ocyphaps lophotes) and the Brush Bronzewing (Phaps elegans) is generally similar to that expected for birds of their size, but the Crested Pigeon has a number of characteristics which would aid survival in hot and dry regions. Body temperature increased similarly for the Crested Pigeon (from 38.8 degrees C to 41.5 degrees C) and the Brush Bronzewing (39.3 degrees C to 41.4 degrees C) over ambient temperatures (T(a)s) from 10 degrees C to 35 degrees C. Both species became hyperthermic (body temperature, T(b)>42 degrees C) at T(a)=45 degrees C. Basal metabolic rate of the Crested Pigeon (0.65 ml O(2) g(-1) h(-1) at 40 degrees C) was approximately 71% of that predicted for a columbid bird, while BMR of the Brush Bronzewing (0.87 ml O(2) g( 1) h(-1) at 20 degrees C to 40 degrees C) was approximately 102% of predicted. Total evaporative water loss increased exponentially with T(a) for both species, from <1 mg H(2)O g(-1) h(-1) at 10 degrees C to >12 mg H(2)O g(-1) h(-1) at 45 degrees C. It was similar and low for both species at T(a)<30 degrees C, but was higher for the Brush Bronzewing than the Crested Pigeon at T(a)>30 degrees C. Ventilatory minute volume matched oxygen consumption, such that oxygen extraction efficiency did not change with T(a) and was similar for both species (approximately 20%). Expired air temperature was considerably lower than T(b) for both species at T(a)<35 degrees C, potentially reducing respiratory water loss by approximately 65% at T(a)=10 degrees C to approximately 30% at T(a)=35 degrees C. Cutaneous evaporative cooling was significant for both species, with skin resistance decreasing as T(a) increased. The Crested Pigeon had a lower skin resistance than the Brush Bronzewing at T(a)=45 degrees C. The Brush Bronzewing had apparently reached its maximum cutaneous water loss at 30 degrees C and relied on panting to cool at higher T(a). PMID- 12743725 TI - Thermoenergetics of pre-moulting and moulting kookaburras (Dacelo novaeguineae): they're laughing. AB - We examined the effect of temperature on resting metabolic rate in seven field captured laughing kookaburras (Dacelo novaeguineae) during late winter and early spring. Basal metabolic rate averaged 201+/-3.4 ml O(2) h(-1) (0.603 ml O(2) g( 1) h(-1)). Overall thermal conductance (K(o)) declined with ambient temperature ( T(a)) and averaged 0.026 ml O(2) g(-1) h(-1) degrees C(-1) at T(a)s<10 degrees C. Day-night differences in body temperatures (2.6 degrees C) and in alpha-phase versus rho-phase minimum metabolic rates were much greater (33%) than predicted for 340-g nonpasserine birds and suggest that these animals operate as low metabolic intensity animals in their rest phase, but normal-metabolic intensity animals during their active phase. Metabolic rate was measured in four of the same birds undergoing moult. Thermal conductance increased to 60% above pre-moult values about 6 weeks after moult began. Basal metabolic rate of moulting birds showing peak thermal conductance readings averaged 17 ml O(2) h(-1) higher than pre-moult measurements. Although this increase was not statistically significant, we believe the moult costs of kookaburras are too low to overcome the inherent variability of BMR determination. We suggest that moult costs of kookaburras are only somewhat higher than the measured costs of protein synthesis of other endotherms. PMID- 12743726 TI - Temperature-activity relationship for the intestinal Na+-K+-ATPase of Sparus aurata. A role for the phospholipid microenvironment? AB - The temperature dependence for Na(+)-K(+)-ATPase has been examined in the proximal-distal axis of the intestine of gilthead seabream (Sparus aurata), i.e. pyloric caeca (PC), anterior intestine (AI) and posterior intestine (PI). Data derived from the Arrhenius plots showed differences in terms of temperature discontinuity points ( Td) (13.29 degrees C, 16.39 degrees C and 17.48 degrees C for PC, AI and PI, respectively) and activation energy ratios (Ea(2)/Ea(1)) obtained at both sides of Td (2.38, 1.98 and 1.78, for PC, AI and PI, respectively). The analyses of polar lipids showed differences in the levels of certain fatty acids among intestinal regions. The content of each fatty acid and different fatty acid ratios were correlated with the corresponding Td and Ea(2)/Ea(1) values. Regression analyses revealed the existence of strong negative correlations between docosahexaenoic acid (22:6n-3, DHA) or the DHA/monoenes ratio and Td. No obvious relationships were observed for other polyunsaturated fatty acids (PUFA) nor saturated fatty acids. The results obtained in the present study indicate that the heterogeneous values of Td displayed by the Na(+)-K(+) ATPase along the intestinal tract could be related to a modulatory role of certain fatty acid within the lipid microenvironment of the enzyme. PMID- 12743727 TI - The influence of dehydration on the thermal preferences of the Western tiger snake, Notechis scutatus. AB - Temperature selection in tiger snakes (Notechis scutatus) is strongly influenced by hydration state and this response varies between two distinct neighbouring populations on semi-arid Carnac Island and mainland wetland (Herdsman Lake). Fed and hydrated (control) Carnac Island snakes selected a preferred body temperature of 26.2+/-1.2 degrees C and an average maximum temperature of 32.5+/-0.5 degrees C in a photo-thermal gradient. Dehydrated Carnac Island snakes selected a significantly lower preferred body temperature (19.7+/-1.6 degrees C) and average maximum temperature (27.7+/-1.0 degrees C). Control Herdsman Lake snakes selected a preferred body temperature of 27.5+/-0.6 degrees C and an average maximum temperature of 33.3+/-0.4 degrees C. Dehydrated Herdsman Lake snakes selected a significantly lower preferred body temperature (23.3+/-1.1 degrees C) and a lower average maximum temperature (31.8+/-0.6 degrees C). Thermal depression (decreased preferred body and average maximum temperatures) in response to dehydration was greater for Carnac Island than Herdsman Lake snakes. As decreases in temperature and activity can reduce water loss, our laboratory data suggest that the survival of the relict population of tiger snakes on Carnac Island is associated with thermoregulatory modifications, which may have the effect of enhancing water conservation in this waterless habitat. PMID- 12743728 TI - High concentrations of isovaleric acid in the fats of odontocetes: variation and patterns of accumulation in blubber vs. stability in the melon. AB - Isovaleric acid (iso5:0) is an unusual fatty acid that is important for echolocation and hearing in acoustic tissues of some odontocetes, but its functional significance in blubber is unknown. We examined patterns of accumulation of this compound in blubber in 30 species of odontocetes ( n=299). Iso5:0 concentrations in blubber varied with phylogeny, ontogeny and body topography. Iso5:0 accumulated in greater quantities in superficial/outer blubber than in deep/inner blubber. In the outer blubber of northern right whale and Hector's dolphins, iso5:0 accounted for one-third to one-half of all fatty acids. Total blubber burden of iso5:0 in harbour porpoises represented up to 15 times the amount deposited in the melon. The composition of the melon does not change during starvation in harbour porpoises, supporting the hypothesis that lipids in melon are conserved for a specific function. Some odontocetes continually deposit iso5:0 in blubber after levels in melon have reached asymptotic levels, suggesting independent control of iso5:0 synthesis and storage in these compartments. Dolphins and porpoises inhabiting cold waters possess higher concentrations of iso5:0 in their outer blubber layers than species from warmer regions. We propose that this relationship represents an adaptive secondary role for iso5:0 in maintaining blubber flexibility in cold environments. PMID- 12743729 TI - Evolutionary aspects of bat echolocation. AB - This review is yet another attempt to explain how echolocation in bats or bat like mammals came into existence. Attention is focused on neuronal specializations in the ascending auditory pathway of echolocating bats. Three different mechanisms are considered that may create a specific auditory sensitivity to echos: (1). time-windows of enhanced echo-processing opened by a corollary discharge of neuronal vocalization commands; (2). differentiation and expansion of ensembles of combination-sensitive neurons in the midbrain; and (3). corticofugal top-down modulations. The second part of the review interprets three different types of echolocation as adaptations to ecological niches, and presents the sophisticated cochlear specializations in constant-frequency/frequency modulated bats as a case study of finely tuned differentiation. It is briefly discussed how a resonant mechanism in the inner ear of constant frequency/frequency-modulated bats may have evolved in common mammalian cochlea. PMID- 12743730 TI - Modeling of time disparity detection by the Hodgkin-Huxley equations. AB - Phase-sensitive neurons in the electrosensory lateral line lobe in the electrosensory pathway of the wave-type electric fish, Gymnarchus niloticus, are specialized for sensing the time disparity between sensory inputs at different parts of the body surface that is necessary for an electrical behavior, jamming avoidance response. These neurons are sensitive to time disparity in the microsecond range between synaptic inputs that represent occurrence times of electrosensory signals at different areas on the body surface. We showed that an ideal Hodgkin-Huxley equation may serve as a time disparity detector that fits physiological precision, and the precision for the time disparity detection is largely regulated by the maximal g(K) conductance in the Hodgkin-Huxley equations. PMID- 12743731 TI - Olfactory sensitivity for aliphatic aldehydes in squirrel monkeys and pigtail macaques. AB - Using a conditioning paradigm, the olfactory sensitivity of three squirrel monkeys and three pigtail macaques for a homologous series of aliphatic aldehydes ( n-butanal to n-nonanal) was assessed. With only few exceptions, the animals of both species significantly discriminated concentrations below 1 ppm from the odorless solvent, and with n-butanal and n-hexanal individual pigtail macaques even demonstrated thresholds below 1 ppb. The results showed (1). both primate species to have a well-developed olfactory sensitivity for aliphatic aldehydes, (2). pigtail macaques to generally perform better than squirrel monkeys in detecting members of this class of odorants, and (3). no significant correlation between perceptibility in terms of olfactory detection thresholds and carbon chain length of the aliphatic aldehydes in both species tested. These findings lend further support to the growing body of evidence suggesting that between species comparisons of the number of functional olfactory receptor genes or of neuroanatomical features are poor predictors of olfactory performance. Further, our findings suggest that olfaction may play an important and hitherto underestimated role in the regulation of behavior in the species tested. PMID- 12743732 TI - Cyclic AMP modulates electrical signaling in a weakly electric fish. AB - Many species of electric fish show diurnal or socially elicited variation in electric organ discharge amplitude. In Sternopygus macrurus, activation of protein kinase A by 8-bromo-cAMP increases electrocyte sodium current magnitude. To determine whether the behavioral plasticity in electric organ discharge amplitude is controlled by electrocyte biophysical properties, we examined whether the effects of phosphorylation on ion currents in the electric organ translate directly into electric organ discharge changes. We injected the electric organ of restrained fish with 8-bromo-cAMP and monitored the electric organ discharge. The effect of protein kinase A activation on electrocyte action potentials was examined in isolated electric organ using two-electrode current clamp. Electric organ discharge and action potential amplitude and pulse duration increased in response to 8-bromo-cAMP. Pulse and action potential duration both increased by about 25%. However, the increase in electric organ discharge amplitude (approximately 400%) was several-fold greater than the action potential amplitude increase (approximately 40%). Resting membrane resistance decreased in electrocytes exposed to 8-bromo-cAMP. We propose that in the Thevenin equivalent circuit of the electric organ a moderate increase in action potential amplitude combined with a decrease in internal resistance produces a greater voltage drop across the external resistance (the water around the fish), accounting for the large increase in the externally recorded electric organ discharge. PMID- 12743733 TI - Sound-producing mechanisms and recordings in Carapini species (Teleostei, Pisces). AB - Carapus boraborensis, C. homei and Encheliophis gracilis are three species of Carapidae that display the ability to penetrate and reside in the holothurian Bohadschia argus. This study describes both the particular morphology of the sound-producing structures and, for the first time, the sounds produced by each species. The study of the structures composing the sound-producing system seems to indicate that the action made by the primary sonic muscles (i.e. the pulling and releasing of the front of the swim bladder) might be responsible for the sound emissions of these three species by provoking a vibration of a thinner zone in front of the swim bladder (swimbladder fenestra). The sounds were only emitted and recorded when several individuals of the same species were inside the same sea cucumber. They were composed of serially repeated knocks and were heard as drum beats or drum rolls. Their specific differences were mainly defined as variations in the timing or grouping of the knocking sounds. The recordings of these sound productions demonstrate a vocal ability for the three species, linked with the presence of particular organs associated with sound production. Moreover, the ecological significance of the sounds and of the sound apparatus system is discussed. PMID- 12743734 TI - Olfactory activation patterns in the antennal lobe of the sphinx moth, Manduca sexta. AB - The sphinx moth Manduca sexta is a well-studied insect with regard to central olfactory functions. Until now, the innervation patterns of olfactory receptor neurons into the array of olfactory glomeruli in the antennal lobe have, however, been unclear. Using optical imaging to visualize calcium dynamics within the antennal lobe we demonstrate specific patterns elicited by sex pheromone components and plant-derived odours. These patterns mainly reflect receptor neuron activity. Within the male-specific macroglomerular complex the two major pheromone components evoke stereotyped activity in either of two macroglomerular complex glomeruli. Based on previous knowledge of output neuron specificity, our results suggest a matching of information between input and output in the macroglomerular complex. Plant odours evoked activity in the sexually isomorphic glomeruli. Two major results were obtained: (1). terpenes and aromatic compounds activate different clusters of glomeruli with only minor overlapping, and (2). the position of certain key glomeruli is fixed in both males and females, which suggests that host-plant related odorants are processed in a similar way in both sexes. PMID- 12743735 TI - Application of real-time polymerase chain reaction technology to detect prostatic bacteria in patients with chronic prostatitis/chronic pelvic pain syndrome. AB - To investigate the potential association between prostate infection and chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS), we used molecular approaches described in previous reports. These methods employed standard polymerase chain (PCR) reaction assays to provide a qualitative evaluation of prostatic bacterial species. Here, we report on the detection of prostatic bacteria using a real-time PCR. Template DNAs were examined from prostatic tissue samples from patients with CP/CPPS. Two PCR primer sets were used: one that amplifies a portion of all known bacterial ribosomal DNAs (16S rDNAs) and one that is specific for Escherichia coli as opposed to related, E. coli-like bacteria. The 16S rDNA real-time PCR assay detected bacterial DNAs in eight (26%) of 31 samples from patients with CP/CPPS, including three samples (10%) that were also positive by the E. coli real-time PCR assay. These E. coli positives were quantified at approximately 10(3) cfu/ml of tissue digested. Quantification, speed and specificity make real time PCR a promising approach for the quantitative detection and identification of prostatic bacteria from CP/CPPS patients. PMID- 12743736 TI - Detection of pulmonary nodules by multislice computed tomography: improved detection rate with reduced slice thickness. AB - The purpose of this study was to find out if the use of 1.25-mm collimated thin slice technique helps to detect more small pulmonary lung nodules than the use of 5 mm. A total of 100 patient examinations that allowed a reconstruction of 1.25 mm slice thickness in addition to the standard of 5-mm slices were included in a prospective study. Acquisition technique included four rows of 1-mm slices. Two sets of contiguous images were reconstructed and compared with 1.25- and 5-mm slice thickness, respectively. Two radiologists performed a film-based analysis of the images. The size and the confidence of the seen nodules were reported. We did not perform a histological verification, according to the normal clinical procedure, although it would be optimal regarding research. Statistical analysis was performed by using longitudinal analysis described by Brunner and Langer. In addition, sensitivity, specificity, negative predictive value and positive predictive value were calculated for each reader using the 1.25-mm sections as the gold standard. As an index for concordance the kappa value was used. A value of p<0.05 was regarded as significant. In 37 patients pulmonary nodules were detected. Twenty-four patients showed more than one nodule; among these, 7 patients had disseminated disease and were excluded from the study. Pulmonary nodules larger than 10 mm in size were equally well depicted with both modalities, whereas lesions smaller than 5 mm in size were significantly better depicted with 1.25 mm (p<0.05). Using 1.25 mm as the gold standard, sensitivity for 5-mm reconstruction interval was 88 and 86% for observers A and B, respectively. No false-positive results were reported for 5-mm sections. Interobserver agreement for nodule detection determined for 1.25-mm reconstruction intervals showed a k value of 0.753, indicating a good agreement, and 0.562 for 5-mm reconstruction intervals, indicating a moderate agreement. Brunner and Langer analysis showed significant differences for slice thickness and no significant difference between the observers. Reduced slice thickness demonstrated an improvement of small nodule detection, confidence levels, and interobserver agreement. Application of thin-slice multidetector-row CT may raise the sensitivity for lung nodule detection, although the higher detection rate of smaller nodules has to be evaluated from a clinical perspective and remains problematic about how the detection of small nodules will effect patient outcome. PMID- 12743737 TI - Sequence analysis and functional characterization of the dialkylglycine decarboxylase gene DGD1 from Mycosphaerella graminicola. AB - Dialkylglycine decarboxylase is a pyridoxal phosphate-dependent enzyme in the aminotransferases class III group of enzymes. The enzyme is unique in terms of catalyzing both decarboxylation and transamination. Although the enzymatic activity is present in some bacteria and fungi, the biological role is unclear. We identified and disrupted the dialkylglycine decarboxylase-encoding gene DGD1 in the wheat blotch fungus Mycosphaerella graminicola by transposon-arrayed gene knockout. The DGD1 gene is highly similar to dialkylglycine decarboxylase from the soil bacterium Burkholderia cepacia. Phylogenetic analysis of various class III aminotransferases showed that dialkylglycine decarboxylases from bacteria and fungi are found in a distinct cluster. Functional analysis revealed that dgd1 disruption mutants display wild-type morphology and pathogenicity to wheat. The dgd1 mutants cannot utilize 2-methylalanine as a sole nitrogen source, as assessed by large-scale nutritional utilization analysis. This is the first description of a mutant phenotype of the fungal dialkylglycine decarboxylase gene. PMID- 12743738 TI - Phase I and pharmacokinetic study of carboplatin and paclitaxel with a biweekly schedule in patients with advanced non-small-cell lung cancer. AB - PURPOSE: A phase I study was conducted to determine the maximum tolerated dose (MTD) and dose-limiting toxicity (DLT) of carboplatin in combination with paclitaxel using a biweekly schedule in patients with advanced non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: The pharmacokinetics of paclitaxel were determined preliminarily in some patients. The criteria for eligibility for study entry included histologically and/or cytologically confirmed NSCLC (stage IIIb or IV), no prior treatment, and measurable disease. Paclitaxel was given in combination with a fixed dose of carboplatin at an area under the concentration time curve (AUC) of 3 mg/ml x min, every 2 weeks. The starting dose of paclitaxel was 100 mg/m(2), and the dose was increased in increments of 20 mg/m(2). Three to six patients were allocated to each dose level. RESULTS: A total of 19 patients (11 male and 8 female) with a median age of 61 years (range 43-74 years) and a median ECOG performance status of 0 (range 0-1) were enrolled. The MTD of paclitaxel proved to be 160 mg/m(2), and the DLT was neutropenia, which improved well following treatment with G-CSF. Gastrointestinal toxicity was well tolerated. Of 17 patients who received four cycles or more, 7 (41%; 95% confidence interval 18.4-67.1%) responded to this combination therapy. The pharmacokinetics of paclitaxel did not differ from published data. CONCLUSIONS: The recommended dose for phase II study is paclitaxel 140 mg/m(2) with a carboplatin AUC of 3 mg/ml.min. This biweekly regimen is highly effective and acceptable, and the present data indicate that the regimen may be suitable for use on an outpatient basis. PMID- 12743749 TI - Phylogenetic analysis of an anaerobic microbial consortium deiodinating 5-amino 2,4,6-triiodoisophthalic acid. AB - The dehalogenating performance of an anaerobic 5-amino-2,4,6-triiodoisophthalic acid (ATIA) fixed-bed reactor was evaluated. The reactor operating conditions were set for ATIA deiodination. A phylogenetic survey for a stable anaerobic ATIA deiodinating microbial consortium was carried out using 16S rDNA restriction fragment length polymorphism, and unique clones were sequenced. Four phylotypes were identified. Two sequences were related to those of Desulfitobacterium frappieri species and another was closest to that of Desulfitobacterium hafniense, but may have represented a new Desulfitobacterium species. Desulfitobacteria were previously described as aryl-dechlorinating and debrominating bacteria. The new strains identified in this study were probably responsible for the ATIA deiodination. The fourth clone was related to the Clostridium-Flavobacterium-Bacteroides group. PMID- 12743750 TI - The treatment of gaseous benzene by two-phase partitioning bioreactors: a high performance alternative to the use of biofilters. AB - A 2-l (1-l working volume) two-phase partitioning bioreactor (TPPB) was used as an integrated scrubber/bioreactor in which the removal and destruction of benzene from a gas stream was achieved by the reactor's organic/aqueous liquid contents. The organic solvent used to trap benzene was n-hexadecane, and degradation of benzene was achieved in the aqueous phase using the bacterium Alcaligenes xylosoxidans Y234. A gas stream with a benzene concentration of 340 mg l(-1) at a flow rate of 0.414 l h(-1) was delivered to the system at a loading capacity of 140 g m(-3) h(-1), and an elimination capacity of 133 g m(-3 )h(-1) was achieved (the volume in this term is the total liquid volume of the TPPB). This elimination capacity is between 3 and 13 times greater than any benzene elimination achieved by biofiltration, a competing biological air treatment strategy. It was also determined that the evaluation of TPPB performance in terms of elimination capacity should include the cell mass present in the system, as this is a readily controllable quantity. A specific benzene utilization rate of 0.57 g benzene (g cells)(-1) h(-1) was experimentally determined in a bioreactor with a cell concentration that varied dynamically between 0.2 and 1 g l(-1). If it assumed that this specific benzene utilization rate (0.57 g g(-1) h(-1)) is independent of cell concentration, then a TPPB operated at high cell concentrations could potentially achieve elimination capacities several hundred times greater than those obtained with biofilters. PMID- 12743751 TI - Control of Lactobacillus contaminants in continuous fuel ethanol fermentations by constant or pulsed addition of penicillin G. AB - The addition of penicillin G to combat microbial contamination in continuous fuel alcohol fermentations was performed using both continuous and pulsed addition regimes. In continuous fermentations where both Saccharomyces cerevisiae and Lactobacillus paracasei were present, the mode of addition of penicillin G determined final numbers of viable L. paracasei. When the same overall average concentration of penicillin G was added in both pulsed and continuous modes, the initial viable number of L. paracasei (8.0 x 10(9) cfu ml(-1)) decreased to a greater degree (1.02 x 10(5) cfu ml(-1) L. paracasei) when penicillin G was pulsed at 6 h frequencies at an overall average concentration of 2,475 U/l than when penicillin G was added continuously at 2,475 U/l (2.77 x 10(5) cfu ml(-1) L. paracasei). Pulsed additions over longer frequencies at 2,475 U/l were not as effective in reducing viable bacteria. Viable yeasts increased during both treatment conditions by more than 2-fold. The two addition regimes also eliminated the 40% decrease in ethanol concentration caused by the intentional bacterial infection. Although there was 3 times more bacterial death with 6 h pulsed additions compared to continuous additions of penicillin G at 2,475 U/l, there was, by that point, no practical difference in either final ethanol concentration or relative ethanol recovery. PMID- 12743753 TI - The Corynebacterium glutamicum genome: features and impacts on biotechnological processes. AB - Corynebacterium glutamicum has played a principal role in the progress of the amino acid fermentation industry. The complete genome sequence of the representative wild-type strain of C. glutamicum, ATCC 13032, has been determined and analyzed to improve our understanding of the molecular biology and physiology of this organism, and to advance the development of more efficient production strains. Genome annotation has helped in elucidation of the gene repertoire defining a desired pathway, which is accelerating pathway engineering. Post genome technologies such as DNA arrays and proteomics are currently undergoing rapid development in C. glutamicum. Such progress has already exposed new regulatory networks and functions that had so far been unidentified in this microbe. The next goal of these studies is to integrate the fruits of genomics into strain development technology. A novel methodology that merges genomics with classical strain improvement has been developed and applied for the reconstruction of classically derived production strains. How can traditional fermentation benefit from the C. glutamicum genomic data? The path from genomics to biotechnological processes is presented. PMID- 12743754 TI - 2-(2'-Hydroxyphenyl)benzene sulfinate desulfinase from the thermophilic desulfurizing bacterium Paenibacillus sp. strain A11-2: purification and characterization. AB - 2-(2'-Hydroxyphenyl)benzene sulfinate (HPBSi) desulfinase (TdsB), which catalyzes the final step of desulfurization of dibenzothiophene (DBT), was purified from a thermophilic DBT- and benzothiophene (BT)-desulfurizing bacterium: Paenibacillus sp. strain A11-2. The molecular mass of the purified enzyme was 31 kDa and 39 kDa by gel filtration and sodium dodecyl sulfate polyacrylamide gel electrophoresis, respectively, suggesting a monomeric structure. The optimal temperature and pH for the reaction involving TdsB was 55 degrees C and the enzyme was more resistant to heat treatment than DszB, a counterpart purified from Rhodococcus erythropolis. The optimum pH for TdsB activity was pH 8. TdsB converted HPBSi to 2-hydroxybiphenyl (2-HBP) and sulfite stoichiometrically. The Km and kcat values for HPBSi were 0.33 mM and 0.32 s(-1), respectively. TdsB was inactivated by SH reagents such as p-chloromercuribenzoic acid and 5,5'-dithio-bis-2-nitrobenzoic acid, but was not inhibited by chelating reagents such as EDTA and o phenanthroline. TdsB was also inhibited by o-hydroxystyrene, the final desulfurized product of BT. However, 2-HBP and its derivatives showed only a weak inhibitory effect. TdsB desulfurized 2-(2'-hydroxyphenyl)ethen-1-sulfinate to yield o-hydroxystyrene, but DszB could not. A site-directed mutagenesis study revealed the cysteine residue at position 17 to be essential to the catalytic activity of TdsB. PMID- 12743752 TI - Overproduction of the Aspergillus niger feruloyl esterase for pulp bleaching application. AB - A well-known industrial fungus for enzyme production, Aspergillus niger, was selected to produce the feruloyl esterase FAEA by homologous overexpression for pulp bleaching application. The gpd gene promoter was used to drive FAEA expression. Changing the nature and concentration of the carbon source nature (maltose to glucose; from 2.5 to 60 g l(-1)), improved FAEA activity 24.5-fold and a yield of 1 g l(-1) of the corresponding protein in the culture medium was achieved. The secreted FAEA was purified 3.5-fold to homogeneity in a two-step purification procedure with a recovery of 69%. The overproduced protein was characterised and presented properties in good agreement with those of native FAEA. The recombinant FAEA was tested for wheat straw pulp bleaching, with or without a laccase mediator system and xylanase. Best results were obtained using a bi-sequential process with a sequence including xylanase, FAEA and laccase, and yielded very efficient delignification--close to 75%--and a kappa number of 3.9. This is the first report on the potential application of recombinant FAEA in the pulp and paper sector. PMID- 12743755 TI - Biotransformation of limonene by bacteria, fungi, yeasts, and plants. AB - The past 5 years have seen significant progress in the field of limonene biotransformation, especially with regard to the regiospecificity of microbial biocatalysts. Whereas earlier only regiospecific biocatalysts for the 1,2 position (limonene-1,2-diol) and the 8-position (alpha-terpineol) were available, recent reports describe microbial biocatalysts specifically hydroxylating the 3 position (isopiperitenol), 6-position (carveol and carvone), and 7-position (perillyl alcohol, perillylaaldehyde, and perillic acid). The present review also includes the considerable progress made in the characterization of plant P-450 limonene hydroxylases and the cloning of the encoding genes. PMID- 12743756 TI - Aminoacyl-tRNA synthetases and their inhibitors as a novel family of antibiotics. AB - The emergence of multidrug-resistant strains of pathogenic microorganisms and the slow progress in new antibiotic development has led in recent years to a resurgence of infectious diseases that threaten the well-being of humans. The result of many microorganisms becoming immune to major antibiotics means that fighting off infection by these pathogens is more difficult. The best strategy to get around drug resistance is to discover new drug targets, taking advantage of the abundant information that was recently obtained from genomic and proteomic research, and explore them for drug development. In this regard, aminoacyl-tRNA synthetases (ARSs) provide a promising platform to develop novel antibiotics that show no cross-resistance to other classical antibiotics. During the last few years there has been a comprehensive attempt to find the compounds that can specifically target ARSs and inhibit bacterial growth. In this review, the current status in the development of ARS inhibitors will be briefly summarized, based on their chemical structures and working mechanisms. PMID- 12743757 TI - Triacylglycerol biosynthesis in yeast. AB - Triacylglycerol (TAG) is the major storage component for fatty acids, and thus for energy, in eukaryotic cells. In this mini-review, we describe recent progress that has been made with the yeast Saccharomyces cerevisiae in understanding formation of TAG and its cell biological role. Formation of TAG involves the synthesis of phosphatidic acid (PA) and diacylglycerol (DAG), two key intermediates of lipid metabolism. De novo formation of PA in yeast as in other types of cells can occur either through the glycerol-3-phosphate- or dihydroxyacetone phosphate-pathways-each named after its respective precursor. PA, formed in two steps of acylation, is converted to DAG by phosphatidate phosphatase. Acylation of DAG to yield TAG is catalyzed mainly by the two yeast proteins Dga1p and Lro1p, which utilize acyl-CoA or phosphatidylcholine, respectively, as acyl donors. In addition, minor alternative routes of DAG acylation appear to exist. Endoplasmic reticulum and lipid particles (LP), the TAG storage compartment in yeast, are the major sites of TAG synthesis. The interplay of these organelles, formation of LP, and enzymatic properties of enzymes catalyzing the synthesis of PA, DAG, and TAG in yeast are discussed in this communication. PMID- 12743758 TI - Biodegradation of microbial and synthetic polyesters by fungi. AB - A variety of biodegradable polyesters have been developed in order to obtain useful biomaterials and to reduce the impact of environmental pollution caused by the large-scale accumulation of non-degradable waste plastics. Polyhydroxyalkanoates, poly(epsilon-caprolactone), poly( l-lactide), and both aliphatic and aromatic polyalkylene dicarboxylic acids are examples of biodegradable polyesters. In general, most aliphatic polyesters are readily mineralized by a number of aerobic and anaerobic microorganisms that are widely distributed in nature. However, aromatic polyesters are more resistant to microbial attack than aliphatic polyesters. The fungal biomass in soils generally exceeds the bacterial biomass and thus it is likely that fungi may play a considerable role in degrading polyesters, just as they predominantly perform the decomposition of organic matter in the soil ecosystem. However, in contrast to bacterial polyester degradation, which has been extensively investigated, the microbiological and environmental aspects of fungal degradation of polyesters are unclear. This review reports recent advances in our knowledge of the fungal degradation of microbial and synthetic polyesters and discusses the ecological importance and contribution of fungi in the biological recycling of waste polymeric materials in the biosphere. PMID- 12743759 TI - Immobilization of xylan-degrading enzymes from Melanocarpus albomyces IIS 68 on the smart polymer Eudragit L-100. AB - Xylanase of Melanocarpus albomyces IIS 68 was immobilized on Eudragit L-100. The latter is a copolymer of methacrylic acid and methyl methacrylate and is a pH sensitive smart polymer. The immobilization was carried out by gentle adsorption and an immobilization efficiency of 0.82 was obtained. The enzyme did not leach off the polymer even in the presence of 1 M NaCl and 50% ethylene glycol. The K(m) of the enzyme changed from 5.9 mg ml(-1) to 9.1 mg ml(-1) upon immobilization. The V(max) of the immobilized enzyme showed an increase from 90.9 micro mol ml(-1) min(-1) (for the free enzyme) to 111.1 micro mol ml(-1) min(-1). The immobilized enzyme could be reused up to ten times without impairment of the xylanolytic activity. The immobilized enzyme was also evaluated for its application in pre-bleaching of eucalyptus kraft pulp. PMID- 12743760 TI - Identifiability and retrievability of unique parameters describing intrinsic Andrews kinetics. AB - A key factor contributing to the variability in the microbial kinetic parameters reported from batch assays is parameter identifiability, i.e., the ability of the mathematical routine used for parameter estimation to provide unique estimates of the individual parameter values. This work encompassed a three-part evaluation of the parameter identifiability of intrinsic kinetic parameters describing the Andrews growth model that are obtained from batch assays. First, a parameter identifiability analysis was conducted by visually inspecting the sensitivity equations for the Andrews growth model. Second, the practical retrievability of the parameters in the presence of experimental error was evaluated for the parameter estimation routine used. Third, the results of these analyses were tested using an example data set from the literature for a self-inhibitory substrate. The general trends from these analyses were consistent and indicated that it is very difficult, if not impossible, to simultaneously obtain a unique set of estimates of intrinsic kinetic parameters for the Andrews growth model using data from a single batch experiment. PMID- 12743761 TI - Studies on a thermostable alpha-amylase from the thermophilic fungus Scytalidium thermophilum. AB - An alpha-amylase produced by Scytalidium thermophilum was purified using DEAE cellulose and CM-cellulose ion exchange chromatography and Sepharose 6B gel filtration. The purified protein migrated as a single band in 6% PAGE and 7% SDS PAGE. The estimated molecular mass was 36 kDa (SDS-PAGE) and 49 kDa (Sepharose 6B). Optima of pH and temperature were 6.0 and 60 degrees C, respectively. In the absence of substrate the purified alpha-amylase was stable for 1 h at 50 degrees C and had a half-life of 12 min at 60 degrees C, but was fully stable in the presence of starch. The enzyme was not activated by several metal ions tested, including Ca(2+) (up to 10 mM), but HgCl(2 )and CuCl(2) inhibited its activity. The alpha-amylase produced by S. thermophilum preferentially hydrolyzed starch, and to a lesser extent amylopectin, maltose, amylose and glycogen in that order. The products of starch hydrolysis (up to 6 h of reaction) analyzed by thin layer chromatography, showed oligosaccharides such as maltotrioses, maltotetraoses and maltopentaoses. Maltose and traces of glucose were formed only after 3 h of reaction. These results confirm the character of the enzyme studied to be an alpha-amylase (1,4-alpha-glucan glucanohydrolase). PMID- 12743762 TI - Significantly enhanced stability of glucose dehydrogenase by directed evolution. AB - An NaCl-independent stability-enhanced mutant of glucose dehydrogenase (GlcDH) was obtained by using in vitro directed evolution. The family shuffling method was applied for in vitro directed evolution to construct a mutant library of GlcDH genes. Three GlcDH-coding genes from Bacillus licheniformis IFO 12200, Bacillus megaterium IFO 15308 and Bacillus subtilis IFO 13719 were each cloned by direct PCR amplification into the p Trc99A expression vector and expressed in the host, Escherichia coli. In addition to these three GlcDH genes, a gene encoding a previously obtained GlcDH mutant, F20 (Q252L), derived from B. megaterium IWG3, was also subjected to directed evolution by the family shuffling method. A highly thermostable mutant, GlcDH DN-46, was isolated in the presence or absence of NaCl after the second round of family shuffling and filter-based screening of the mutant libraries. This mutant had only one novel additional amino acid residue exchange (E170K) compared to F20, even though DN-46 was obtained by family shuffling of four different GlcDH genes. The effect of temperature and pH on the stability of the GlcDH mutants F20 and DN46 was investigated with purified enzymes in the presence or absence of NaCl. In the absence of NaCl, F20 showed very poor thermostability (half-life =1.3 min at 66 degrees C), while the half life of isolated mutant DN-46 was 540 min at 66 degrees C, i.e., 415-fold more thermostable than mutant F20. The activity of the wild-type and F20 enzymes dropped critically when the pH value was changed to the alkaline range in the absence of NaCl, but no such decrease was apparent with the DN-46 enzyme in the absence of NaCl. PMID- 12743763 TI - Cloning, expression and characterisation of two tyrosinase cDNAs from Agaricus bisporus. AB - Using primers designed on the basis of sequence homologies in the copper-binding domains for a number of plant and fungal tyrosinases, two tyrosinase encoding cDNAs were cloned from an Agaricus bisporus U1 cDNA-library. The sequences AbPPO1 and AbPPO2 were, respectively, 1.9 and 1.8 kb in size and encoded proteins of approximately 64 kDa. The cDNAs represent different loci. Both AbPPO1 and AbPPO2 occur as single copies on the genomes of the U1 parental strains H39 and H97. The genomic size of AbPPO1 and AbPPO2 is minimally 2.3 and 2.2 kb, respectively. Alignment and phylogenetic analysis of 35 tyrosinase and polyphenol oxidase sequences of animal, plant, fungal, and bacterial origin indicated conserved copper-binding domains, and stronger conservation within genera than between them. The translation products of AbPPO1 and AbPPO2 possess putative N glycosylation and phosphorylation sites and are recognised by antibodies directed against a 43-kDa tyrosinase. The observations are consistent with previously proposed maturation and activation models for plant and fungal tyrosinases. PMID- 12743765 TI - Corn fiber hydrolysis by Thermobifida fusca extracellular enzymes. AB - Thermobifida fusca was grown on cellulose (Solka-Floc), xylan or corn fiber and the supernatant extracellular enzymes were concentrated. SDS gels showed markedly different protein patterns for the three different carbon sources. Activity assays on a variety of synthetic and natural substrates showed major differences in the concentrated extracellular enzyme activities. These crude enzyme preparations were used to hydrolyze corn fiber, a low-value biomass byproduct of the wet milling of corn. Approximately 180 mg of reducing sugar were produced per gram of untreated corn fiber. When corn fiber was pretreated with alkaline hydrogen peroxide, up to 429 mg of reducing sugars were released per gram of corn fiber. Saccharification was enhanced by the addition of beta-glucosidase or by the addition of a crude xylanase preparation from Aureobasidium sp. PMID- 12743764 TI - Biodegradation of phenanthrene by Pseudomonas sp. strain PP2: novel metabolic pathway, role of biosurfactant and cell surface hydrophobicity in hydrocarbon assimilation. AB - Pseudomonas sp. strain PP2 isolated in our laboratory efficiently metabolizes phenanthrene at 0.3% concentration as the sole source of carbon and energy. The metabolic pathways for the degradation of phenanthrene, benzoate and p hydroxybenzoate were elucidated by identifying metabolites, biotransformation studies, oxygen uptake by whole cells on probable metabolic intermediates, and monitoring enzyme activities in cell-free extracts. The results obtained suggest that phenanthrene degradation is initiated by double hydroxylation resulting in the formation of 3,4-dihydroxyphenanthrene. The diol was finally oxidized to 2 hydroxymuconic semialdehyde. Detection of 1-hydroxy-2-naphthoic acid, alpha naphthol, 1,2-dihydroxy naphthalene, and salicylate in the spent medium by thin layer chromatography; the presence of 1,2-dihydroxynaphthalene dioxygenase, salicylaldehyde dehydrogenase and catechol-2,3-dioxygenase activity in the extract; O(2) uptake by cells on alpha-naphthol, 1,2-dihydroxynaphthalene, salicylaldehyde, salicylate and catechol; and no O(2) uptake on o-phthalate and 3,4-dihydroxybenzoate supports the novel route of metabolism of phenanthrene via 1-hydroxy-2-naphthoic acid --> [alpha-naphthol] --> 1,2-dihydroxy naphthalene --> salicylate --> catechol. The strain degrades benzoate via catechol and cis,cis muconic acid, and p-hydroxybenzoate via 3,4-dihydroxybenzoate and 3-carboxy- cis,cis-muconic acid. Interestingly, the culture failed to grow on naphthalene. When grown on either hydrocarbon or dextrose, the culture showed good extracellular biosurfactant production. Growth-dependent changes in the cell surface hydrophobicity, and emulsification activity experiments suggest that: (1) production of biosurfactant was constitutive and growth-associated, (2) production was higher when cells were grown on phenanthrene as compared to dextrose and benzoate, (3) hydrocarbon-grown cells were more hydrophobic and showed higher affinity towards both aromatic and aliphatic hydrocarbons compared to dextrose-grown cells, and (4) mid-log-phase cells were significantly (2-fold) more hydrophobic than stationary phase cells. Based on these results, we hypothesize that growth-associated extracellular biosurfactant production and modulation of cell surface hydrophobicity plays an important role in hydrocarbon assimilation/uptake in Pseudomonas sp. strain PP2. PMID- 12743766 TI - Dual influence of the carbon source and L-methionine on the synthesis of sulphur compounds in the cheese-ripening yeast Geotrichum candidum. AB - The effect of the carbon source and l-methionine on the ability of Geotrichum candidum to produce volatile sulphur compounds (VSC) was studied. This yeast was cultivated in a synthetic medium supplemented with various carbon sources and l methionine at different concentrations. Both glycerol and glucose significantly increased VSC production by G. candidum. Unlike the effect on the l-methionine- and 4-methylthio-2-oxobutyric acid-demethiolating activities, the supply of a carbon source had a dramatic effect on the activity of aminotransferase, a key enzyme in l-methionine catabolism. An increase in the initial concentration of l methionine resulted in a rise in the production of sulphur compounds (VSC, 4 methylthio-2-oxobutyric acid) but had limited effect on l-methionine-catabolising enzyme activities. Evidence for the existence of a dual effect of the carbon source and l-methionine on VSC biosynthesis was obtained in this study. PMID- 12743767 TI - Pseudomonas putida as the dominant toluene-degrading bacterial species during air decontamination by biofiltration. AB - The microbial communities established in three laboratory-scale compost matrix biofilters fed with toluene were characterized. The biofilters were operated for 7 weeks at inlet concentrations of toluene ranging over 250-500 ppm with daily irrigation, using a nutrient solution containing variable concentrations of nitrogen, supplied as urea, and other inorganic salts. The indigenous microflora of the compost included toluene-degrading species, making inoculation unnecessary. The numerically predominant toluene-degrading strains were isolated from the most diluted positive wells of most-probable-number counts on mineral medium with toluene as sole carbon source and identified by rRNA 16S gene sequencing. On the basis of sequence similarity, all the isolated strains were assigned to the species Pseudomonas putida, although some variations were observed in their respective sequences. It is concluded that the mode of biofilter operation including a daily supply of non-carbon nutrients created an environment favoring the constant numerical predominance of this fast-growing toluene-degrading species. PMID- 12743769 TI - Biodegradation of chloronaphthalenes and polycyclic aromatic hydrocarbons by the white-rot fungus Phlebia lindtneri. AB - The biodegradation of chloronaphthalene (CN) and polycyclic aromatic hydrocarbons by the white-rot fungus Phlebia lindtneri, which can degrade dichlorinated dioxins and non-chlorinated dioxin-like compounds, was investigated. Naphthalene, phenanthrene, 1-chloronaphthalene (1-CN) and 2-chloronaphthalene (2-CN) were metabolized by the fungus to form several oxidized products. Naphthalene and phenanthrene were metabolized to the corresponding hydroxylated and dihydrodihydroxylated metabolites. 2-CN was metabolized to 3-chloro-2-naphtol, 6 chloro-1-naphtol and two other chloronaphtols, CN-dihydrodiols and CN-diols. Significant inhibition of the degradation of these substrates was observed when they were incubated with the cytochrome P-450 monooxygenase inhibitors 1 aminobenzotriazole and piperonyl butoxide. These results suggest that P. lindtneri initially oxidizes these substrates by a cytochrome P-450 monooxygenase. PMID- 12743768 TI - Lignin degradation in a compost environment by the deuteromycete Paecilomyces inflatus. AB - Two strains of the deuteromycete Paecilomyces inflatus were isolated from compost samples consisting of municipal wastes, paper and wood chips. Lignin degradation by P. inflatus was studied following the mineralization of a synthetic (14)C(beta)-labeled lignin (side-chain labeled dehydrogenation polymer, DHP). Approximately 6.5% of the synthetic lignin was mineralized during solid-state cultivation of the fungus in autoclaved compost; and 15.5% was converted into water-soluble fragments. Laccase was the only ligninolytic enzyme detectable when the isolates were grown in autoclaved compost. Production of the enzyme was growth-associated and dependent on the culture conditions. The optimal pH for laccase production was between 4.5 and 5.5 and the optimal temperature was around 30 degrees C. Activity levels of laccase increased in the presence of low molecular-mass aromatic compounds, such as veratryl alcohol, veratric acid, vanillin and vanillic acid. PMID- 12743770 TI - Chronic pancreatitis in Japan: epidemiology, prognosis, diagnostic criteria, and future problems. AB - Chronic pancreatitis is a chronic clinical disorder characterized by irreversible damage to the pancreas and the development of histologic evidence of inflammation and fibrosis, and eventually the destruction and permanent loss of exocrine and endocrine tissue. A nationwide survey in Japan revealed an increase in the total number of patients treated for chronic pancreatitis from 32 000 in 1994 to 42 000 in 1999. The overall prevalence and the incidence rate of chronic pancreatitis also increased, from 28.5 and 5.4, respectively, in 1994 to 32.91 and 5.77 per 100 000 population, respectively, in 1999. Diagnostic criteria for chronic pancreatitis in Japan were proposed by the Japan Pancreas Society in 1995 and revised in 2001. The criteria were established to rule out false-positive cases and to confidently diagnose definite and probable cases of chronic pancreatitis, and thus easily detect advanced chronic pancreatitis, but the criteria are unable to lead to the early diagnosis of chronic pancreatitis. Cancer is the major cause of death in patients with chronic pancreatitis in Japan (49.6% of all deaths in patients with chronic pancreatitis). Clarification of the mechanisms by which possible chronic pancreatitis progresses to probable or definite chronic pancreatitis, and to pancreatic cancer, is an important research goal. Because even chronic pancreatitis defined as irreversible appears to be reversible for some time in its clinical course, there is an urgent need to develop methods for diagnosing reversible chronic pancreatitis, and to prevent the transition from chronic pancreatitis to pancreatic cancer. PMID- 12743771 TI - Abnormal bile flow in patients with achalasia. AB - BACKGROUND: It has been reported that esophageal achalasia is frequently associated with the dysmotility of other digestive organs. However, the prevalence of extraesophageal complications in patients with achalasia still remains poorly understood. We performed cholescintigraphy, using (99m)Tc pyridoxyl-5-methyl-tryptophan, in patients with esophageal achalasia to assess any possible dysfunction of the sphincter of Oddi associated with achalasia. METHODS: Eight patients (two men and six women) were examined to determine the time required for bile to flow from the bile duct to the duodenum. RESULTS: Excretion time of bile was markedly prolonged in five of the eight patients with achalasia. Scintigraphic findings were not correlated with the radiographic classification of achalasia or with the grading of achalasia. CONCLUSIONS: The present results suggest that a considerable number of patients with achalasia have dysfunction of the sphincter of Oddi, irrespective of the morphological type of achalasia and the grade of esophageal dilatation. PMID- 12743772 TI - Serum pepsinogen concentration as a marker of Helicobacter pyloriinfection and the histologic grade of gastritis; evaluation of gastric mucosa by serum pepsinogen levels. AB - BACKGROUND: Helicobacter pylori infection and associated gastritis are well-known significant factors in many gastrointestinal diseases, and evaluation of these conditions is important for health evaluation. We investigated the utility of serum pepsinogen (PG) concentrations for the diagnosis of H. pylori infection and evaluation of the grade of histologic gastritis. METHODS: The subjects consisted of 283 individuals (147 men and 136 women; mean age, 44.0 years). Biopsy specimens were obtained from the gastric antrum and body to assess grade of inflammation and atrophy and histologic evidence of H. pylori infection. H. pylori infection was judged by Giemsa staining and serum IgG antibodies against H. pylori. PG concentrations were determined by radioimmunoassay. RESULTS: In subjects with H. pylori infection, serum PGII concentrations were increased, and the PGI/PGII ratio (I/II ratio) was decreased. In patients with marked atrophy or intestinal metaplasia, both serum PGI and the I/II ratio were decreased. When PGII concentrations of 12 ng/ml or more, or a I/II ratio of 4.0 or less were used as the cutoff points for the diagnosis of H. pylori infection, the sensitivity and specificity of diagnosis were 90.0% and 93.5%, respectively. All subjects with serum PGI concentrations of 85 or more ng/ml, or serum PGII concentrations of 15 or more ng/ml were H. pylori-positive and all subjects with a I/II ratio of more than 6.5 were H. pylori-negative. CONCLUSIONS: These results suggest that H. pylori infection, gastritis, and glandular atrophy of the stomach can be evaluated via serum PG concentrations, allowing the evaluation of gastric mucosal integrity. PMID- 12743773 TI - Follow-up survey of a large-scale multicenter, double-blind study of triple therapy with lansoprazole, amoxicillin, and clarithromycin for eradication of Helicobacter pylori in Japanese peptic ulcer patients. AB - BACKGROUND: To evaluate histopathological changes and effects on inhibition of ulcer recurrence, a follow-up survey was performed in Japanese patients with Helicobacter pylori-positive active peptic ulcers. These patients had previously participated in a large-scale multicenter trial of triple therapy with lansoprazole (LPZ)/amoxicillin (AMPC)/clarithromycin (CAM) for eradication of H. pylori. METHODS: Patients who had been treated with LPZ only or a combination of LPZ, AMPC, and CAM for a period of 7 days and in whom ulcer healing had been confirmed after treatment were grouped according to successful or failed eradication of H. pylori. They were examined endoscopically to determine whether ulcers had recurred. The updated Sydney system was applied to study histological changes after H. pylori eradication therapy, compared with baseline. RESULTS: Twelve months after treatment for H. pylorieradication, gastric ulcers had recurred in 11.4% of those with successful H. pylorieradication and in 64.5% of those with unsuccessful H. pylori eradication. Duodenal ulcers had recurred in 6.8% of patients for whom H. pylori eradication was successful and in 85.3% of patients in whom eradication failed. These findings proved that H. pylori eradication significantly reduced ulcer recurrence ( P < 0.0001 for both types of ulcers). Histopathological findings of inflammation and activity grade in both gastric and duodenal ulcers were more favorable in patients with successful eradication than in those with unsuccessful eradication. CONCLUSIONS: H. pylori eradication significantly inhibited ulcer recurrence in Japanese peptic ulcer patients. Histopathological findings were also improved with regard to inflammation and activity (neutrophils) in patients in whom H. pylori eradication was successful. PMID- 12743774 TI - The expression of matrix metalloproteinase matrilysin indicates the degree of inflammation in ulcerative colitis. AB - BACKGROUND: Any alteration in the synthesis and breakdown of the extracellular matrix is important in tissue remodeling during inflammation and wound healing. The degradation of the extracellular matrix components is regulated by a cascade of matrix metalloproteinases (MMPs). The present study attempted to assess the relationship between MMPs and the degree of inflammation in ulcerative colitis. METHODS: The expression of MMPs, including MMP-1, -2, -3, -7 (matrilysin), and 9, and that of their inhibitors (tissue inhibitor of metalloproteinases [TIMP]-1 and -2) were analyzed immunohistochemically by using 52 formalin-fixed and paraffin-embedded specimens from patients with ulcerative colitis who had undergone a biopsy or surgery. RESULTS: It was observed that MMP-1, -2, and -9, and the TIMPs were expressed in stromal cells, MMP-3 was expressed in both the epithelial cells and stromal components, and matrilysin was expressed only in the epithelial cells on the edge of ulcers. The expression of the MMPs was increased compared with that of the TIMPs. The frequency of matrilysin expression was increased corresponding to the severity of the inflammation. Matrilysin was also expressed in epithelial cells with dysplasia and cancer. CONCLUSIONS: Matrilysin expression could be an important marker of activity and could be used for the prediction of subsequent transformation in patients with ulcerative colitis. PMID- 12743775 TI - Prevalence of diabetes mellitus in Japanese patients infected chronically with hepatitis C virus. AB - BACKGROUND: To examine the relationship between hepatitis C virus (HCV) infection and diabetes mellitus (DM) in Japanese populations, a retrospective study was done in 866 patients with chronic viral disease. METHODS: The present study included 707 HCV-infected and 159 hepatitis B virus (HBV)-infected patients. The prevalences of HBV- and HCV-related cirrhosis were 32% and 33%, respectively. A case-control study was also conducted to determine the seroprevalence of HCV infection in a cohort of 459 diabetics. RESULTS: The prevalence of DM was higher in HCV-infected patients (20.9%; P < 0.02) than in HBV-infected subjects (11.9%). In the cirrhotic patients, DM was observed in 30.8% of the subjects with HCV compared with 11.8% of those with HBV ( P < 0.01). Multivariate analysis revealed that the major independent variables associated with type II DM were male sex (odds ratio, 1.54; p = 0.020) and cirrhosis (odds ratio, 1.97; P = 0.0007). The relative odds of the development of DM were calculated to be 3.2 times higher in HCV-infected cirrhotic patients than in HBV-infected ones. In the case-control study of the diabetic cohort, 10.5% of patients were infected with HCV compared with 1.1% with HBV ( P < 0.0001). The results indicate that HCV infection is closely associated with DM, compared with HBV infection. Cirrhosis was an independent risk factor for DM. CONCLUSIONS: Taken together, the findings indicate that cirrhosis appears to be a more important predictor of glucose intolerance than HCV infection, and the combination of both factors increases the risk of DM in our populations. PMID- 12743776 TI - Endoscopic injection sclerotherapy for esophageal varices in cirrhotic patients without hepatocellular carcinoma: a comparison of longterm survival between prophylactic therapy and emergency therapy. AB - BACKGROUND: This study aimed to determine whether prophylactic endoscopic injection sclerotherapy prolonged survival in patients with esophageal varices complicated by liver cirrhosis in the absence of hepatocellular carcinoma, compared with emergency sclerotherapy. METHODS: The subjects included 160 patients suffering from esophageal varices complicated by liver cirrhosis without hepatocellular carcinoma. Sixty-eight patients underwent emergency therapy for bleeding varices and the remaining 92 patients underwent prophylactic sclerotherapy. All subjects continued to receive therapy until the varices disappeared. RESULTS: Five-year survival was significantly better in the prophylactic group compared with the emergency group. During the 5-year observation period, 20 of the 68 patients in the emergency group experienced rebleeding and 5 patients died as a result of rebleeding. These rates were significantly higher than those in the prophylactic group (1 of 9 patients with bleeding died among the 92 prophylactic sclerotherapy patients). Multivariate analysis showed that prophylactic therapy and Child's C hepatic function were significant factors for 5-year survival. CONCLUSIONS: Prophylactic sclerotherapy for esophageal varices might be more effective in prolonging longterm survival of patients complicated by liver cirrhosis in the absence of hepatocellular carcinoma, compared with emergency sclerotherapy. PMID- 12743777 TI - Mutational analysis of the pancreatic secretory trypsin inhibitor gene in familial and juvenile pancreatitis in Japan. AB - BACKGROUND: Mutations in the pancreatic secretory trypsin inhibitor ( PSTI) gene have been reported in patients suffering from chronic pancreatitis. The aim of the present study was to investigate whether similar gene mutations are present in familial and juvenile pancreatitis patients in Japan. METHODS: All four exons of the PSTI gene and their flanking intronic sequences were amplified by polymerase chain reaction and sequenced for 37 familial pancreatitis patients (24 families) and 15 juvenile pancreatitis patients, distributed throughout Japan. RESULTS: Three types of exonic mutation in the PSTI gene were observed. The N34S mutation was found in six familial pancreatitis patients (three families) and in one juvenile pancreatitis patient, and the R67C mutation was found in one familial pancreatitis patient and one juvenile pancreatitis patient. We also found a 272C>T mutation in the 3' untranslated region of exon 4 in one familial pancreatitis patient and four juvenile pancreatitis patients. It should be noted that the N34S mutation was cosegregated with two intronic mutations, specifically, IVS1-37T>C and IVS3-69insTTTT. CONCLUSIONS: The same set of N34S mutations (N34S + IVS1-37T>C + IVS3-69insTTTT) that exists in other countries was found in the PSTI gene in Japanese familial and juvenile pancreatitis patients. Another unique mutation (R67C) was also observed in two patients; 272C>T was suggested to be a normal polymorphism. PMID- 12743778 TI - Isolated granulomatous gastritis successfully treated by Helicobacter pylori eradication: a possible association between granulomatous gastritis and Helicobacter pylori. AB - We report two cases of granulomatous gastritis that resolved completely with Helicobacter pylori eradication. Two Japanese women, one 61 years old and one 58 years old, presented to our hospital with epigastralgia. Endoscopy revealed multiple shallow ulcerative lesions in the gastric corpus of both patients. Biopsy specimens demonstrated acute and chronic inflammation with multiple small, noncaseating granulomas at the level of the foveolar isthmi-the junction between the pits and the glands. Both specimens were positive for Helicobacter pylori. No other causes of organic granulomatous disease could be found. Both patients were diagnosed as having isolated granulomatous gastritis and were given triple therapy for Helicobacter pylorieradication, with their fully informed consent. The granulomatous gastritis resolved after successful Helicobacter pylorieradication therapy. This report describes a possible association between isolated granulomatous gastritis and Helicobacter pyloriinfection. To our knowledge, this is the first report describing isolated granulomatous gastritis successfully treated by triple therapy ( Helicobacter pylorieradication therapy). PMID- 12743780 TI - Granular cell tumor occurring in the sigmoid colon treated by endoscopic mucosal resection using a transparent cap (EMR-C). AB - A case of granular cell tumor occurring in the sigmoid colon is reported. The patient, a 56-year-old man, visited our hospital for further evaluation of occult blood in his stool. Endoscopic examination revealed a yellowish, hemispheric submucosal tumor (SMT) with redness, about 6 mm in diameter, in the sigmoid colon. Endoscopic mucosal resection using a transparent cap (EMR-C) was performed, and histological examination revealed that the tumor consisted of a nested growth of large tumor cells with ample granular cytoplasm and small round nuclei. The tumor cells expressed S-100 protein and were stained with neuron specific enolase (NSE) and periodic acid-Schiff, but were negative for desmin, vimentin, and cytokeratin. The resected tumor was diagnosed as a granular cell tumor. This may be the first report of a colorectal granular cell tumor successfully treated with EMR-C. PMID- 12743779 TI - Primary colorectal T-cell lymphoma. AB - We report here a case of primary colorectal T-cell lymphoma in a 49-year-old man. Eighteen years previously, he was diagnosed as having ulcerative colitis based on the findings of colonoscopy and a barium enema. Since then, he had been treated with salicylazosulfapyridine until the most recent episode. He was refered to our clinic with the chief complaint of abdominal pain and excretion of mucus, and for a workup of bowel lesions. Physical examination results were not remarkable, except for the presence of low-grade fever. Total colonoscopy showed multiple shallow ulcers and aphthoid erosions through the entire colon and rectum, except for the descending colon. Endoscopic findings of the descending colon were normal, which was different from the findings of the active stage of ulcerative colitis. Biopsy specimens from the colon and rectum with ulcerations and aphthoid erosions showed a diffuse proliferation of medium-sized to large atypical lymphoid cells with irregular and indistinct nucleoli, thus revealing malignant lymphoma, diffuse pleomorphic type. The lymphoma cells were positive for CD2, CD3, CD5, CD8, and T-cell receptor (TCR) beta F1, but negative for CD4, CD19, CD20, CD103, and CD56. Southern blotting revealed rearrangement of TCR. Based on these findings, the patient was diagnosed as having high-grade T-cell lymphoma. The findings of computerized tomography of the chest and abdomen, gallium scintigraphy, and abdominal ultrasonography were all normal. There were no abdominal lesions throughout the esophagus, stomach, duodenum, and small intestine. As the patient refused total proctocolectomy, he was treated with one course of CHOP (cyclophosphamide, vincristine, adriamycin, and prednisolone) and subsequently with three courses of ProMACE-CytaBOM (consisting of cyclophosphamide, adriamycin, etoposide, cytarabine, bleomycin, vincristine, methotrexate, and prednisolone). After the therapy, improvement of the colorectal lesions was observed, though lesions clearly still remained. To our knowledge, this is the first case report of primary colorectal T-cell lymphoma with cytotoxic/suppressor T-cell phenotype. PMID- 12743781 TI - A decreased number of c- kit-expressing cells in a patient with afferent loop syndrome. AB - Following gastrectomy, stasis in the afferent jejunal loop accompanied by an overgrowth of bacteria leads to a number of clinical symptoms, including the so called afferent loop syndrome. The disturbances in intestinal motility may be related to stagnation of the intestinal contents in the afferent loop. The pacemaker cells for the basic contractile activity of the intestine are thought to be the interstitial cells of Cajal (ICCs). We and others have reported that ICCs express the c- kit receptor, and that a decreased number of c- kit expressing ICCs is generally thought to result in disturbed intestinal motility. We report here a patient with postgastrectomy afferent loop syndrome with a decreased number of c- kit-expressing cells in the external muscle layer of the dilated intestine, suggesting damage to the ICCs. PMID- 12743782 TI - Type C chronic hepatitis with the discovery of a small hepatocellular carcinoma 7 years after successful interferon therapy. AB - The patient, a 61-year-old man, had sustained injuries in a traffic accident at the age of 26, for which he received a blood transfusion. Since 1988 (age, 49 years), abnormal hepatic function had been detected, and, because of the presence of hepatitis C virus antibodies, he was diagnosed as having type C chronic hepatitis. Based on a liver biopsy that was conducted in July 1992 (age, 53), a histological diagnosis of chronic active hepatitis (F(1)/A(2)) was made. Over a period of 6 months, starting in 1992, the patient was treated with interferon (IFNalpha-2a; total dosage, 720 MU). At the end of this regimen, the alanine aminotransferase level was normalized and serum hepatitis C virus-ribonucleic acid was negative. This condition was maintained until August 1996 (age, 57), after which the patient stopped reporting to our hospital. In June 2000 (age, 61) when he was hospitalized for an adhesive ileus, a small hepatocellular carcinoma (a solitary lesion measuring 18 mm in diameter) at S(8) was found, and it was extirpated by a segmental excision in July. The case is introduced to call attention to the need for longterm follow-up observation, even after effective IFN therapy. PMID- 12743783 TI - Secondary hemocholecyst after radiofrequency ablation therapy for hepatocellular carcinoma. AB - A 65-year-old Japanese man underwent radiofrequency ablation (RFA) therapy of a hepatocellular carcinoma. Hemobilia from the intrahepatic bile ducts adjacent to the tumor developed on the fifth day after the RFA therapy. Ultrasonograms and computed tomograms showed swelling of the gallbladder, which was filled with a clot, suggesting the diagnosis of hemocholecyst. The hemobilia resolved with conservative therapy, but a cholecystectomy was performed to manage postprandial abdominal pain. The resected gallbladder was filled with a clot, but injury or ulceration of the gallbladder was absent, suggesting that the hemocholecyst developed secondary to the hemobilia. Secondary hemocholecyst is a rare complication of RFA therapy. The number of cases of secondary hemocholecyst is likely to increase as hepatocentestic therapy becomes more common. Cholecystectomy is indicated for hemocholecyst because spontaneous liquefication and drainage of a clot in the gallbladder usually does not occur. PMID- 12743784 TI - Magnetic resonance imaging in the diagnosis of neurotic abdominal bloating and pain. PMID- 12743785 TI - Stage IV hepatocellular carcinoma with portal venous tumor thrombus: complete response after combined therapy of repeated arterial chemoembolization and radiofrequency ablation. PMID- 12743786 TI - Peptic ulcer recurrence and Helicobacter pylori: evidence from Japan. PMID- 12743787 TI - Mucosal expression of matrix metalloproteinases and their tissue inhibitors in ulcerative colitis patients. PMID- 12743788 TI - Alterations of interstitial cells of Cajal and gastrointestinal motility disorders. PMID- 12743789 TI - Development of hepatocellular carcinoma after longterm sustained complete response to interferon therapy: what is the mechanism? PMID- 12743790 TI - Assessment of nutritional status and prognosis in advanced cancer: interleukin-6, C-reactive protein, and the prognostic and inflammatory nutritional index. By Declan Walsh et al. Supportive Care in Cancer (2003) 11:60-62. PMID- 12743791 TI - Association of high-altitude systemic hypertension with the deletion allele-of the angiotensin-converting enzyme (ACE) gene. AB - People who visit high-altitude areas are exposed to a stressful environment and a good percentage of them suffer from high-altitude-induced diseases, including systemic hypertension. Identification of genetic markers for high-altitude induced diseases would help to reduce the rate of morbidity/mortality from such diseases. The development of systemic hypertension on exposure to high altitude (3,500 m) for 30 days in otherwise normotensive natives of low-altitudes was investigated. The angiotensin-converting enzyme (ACE) insertion/deletion (I/D) genotypes and renin-angiotensin-aldosterone system were simultaneously studied. In the hypertensives during their stay at high altitude, the ACE D allele frequency was significantly higher than in the normotensives (0.67 versus 0.32 chi(2)(1) = 10.6, P < 0.05). In the normotensives during their stay at high altitude, there was no significant increase in plasma aldosterone levels despite increased plasma renin activity. Results of the present study suggest that environmental changes and pre-existing genetic factors, namely the ACE D allele, might be two of the factors predisposing natives of low altitudes to systemic hypertension, a polygenic disease, at high altitude. PMID- 12743792 TI - Successful renal transplantation in a child with ANCA-associated microscopic polyangiitis. AB - Crescentic glomerulonephritis (CGN) is a clinicopathologic entity which is characterized by severe renal dysfunction of rapid onset with glomerular crescents. Type III CGN is associated with the absence of glomerular immune complex deposition (pauci-immune) and is associated with antineutrophil cytoplasmic antibody (ANCA). Microscopic polyangiitis and idiopathic pauci-immune necrotizing glomerulonephritis (NCGN) are strongly associated with ANCA directed against myeloperoxidase (anti-MPO). We describe here an unusual pediatric patient with MPO-ANCA-associated rapidly progressive glomerulonephritis (RPGN), emphasizing the management and outcome of the disease. PMID- 12743793 TI - Histopathological spectrum of childhood nephrotic syndrome in Indian children. AB - Nephrotic syndrome in children is a clinical manifestation of different histopathological subtypes. There is a paucity of recent large studies dealing with the histopathological spectrum from developing countries. A prospective study was performed from January 1990 to December 2000 at our center, involving 600 children (with age of onset up to 16 years) with idiopathic nephrotic syndrome (INS). The objectives were: (1) to study the histopathological distribution of different subtypes of INS and (2) to compare the clinical and biochemical parameters at the time of diagnosis of minimal change disease (MCD) with non-MCD subtypes. For the purpose of this study we analyzed only those children with INS who underwent biopsies. The study group included 290 children in which adequate biopsy reports were available. There were 213 males and 77 females. Mean age at onset of INS was 7.9+5.1 years. Facial edema was found in 286 (98.6%), microhematuria in 120 (41.3%), gross hematuria in 7 (2.5%), and hypertension in 77(26.8%) patients. All patients of the study group were seronegative for HBsAg and HIV. Focal and segmental glomerulosclerosis (FSGS) was the most common histopathological subtype, occurring in 110 of 290 children (38%). Other subtypes included MCD in 95 children (32%), membranoproliferative glomerulonephritis (MPGN) in 44 children (15%), mesangioproliferative glomerulonephritis in 33 children (11%), membranous glomerulonephritis in 5 children (2%), and diffuse mesangial sclerosis in 3 children (1%). In children under 8 years of age, MCD was the most common entity, whereas FSGS predominated in children with age at onset greater than 8 years. The age at onset of nephrotic syndrome was significantly higher in the non-MCD group than the MCD group. The incidence of hypertension, microhematuria, and gross hematuria was significantly lower in the MCD group. MCD remains the most common histopathological subtype in Indian children with INS and onset under 8 years of age. The incidence of MPGN continues to be high. MCD can be differentiated from non-MCD subtype by younger age at onset, absence of hypertension, and absence of microscopic hematuria. PMID- 12743794 TI - Does flood tolerance explain tree species distribution in tropical seasonally flooded habitats? AB - In the tropics, seasonally flooded forests (SFF) harbor fewer tree species than terra firme (i.e. non-flooded) forests. The low species diversity of tropical flooded forests has been ascribed to the paucity of species with adaptations to tolerate flooding. To test the hypothesis that flooding is the only factor restricting most species from SFF, we compared plant morphological and physiological responses to flooding in 2-month old seedlings of 6 species common to SFF and 12 species common to terra firme forests. Although flooding impaired growth, total biomass, maximum root length and stomatal conductance in most species, responses varied greatly and were species-specific. For example, after 90 days, flooding reduced leaf area growth by 10-50% in all species, except in Tabebuia, a common species from non-flooded habitats. Similarly, flooding had a 5 45% negative effect on total biomass for all species, except in 1 SFF and 1 terra firme species both of which had more biomass under flooding. A principal component analysis, using the above responses to flooding, provided no evidence that SFF and terra firme species differed in their responses to flooding. Flooding also caused reductions in root growth for most species. Rooting depth and root: shoot ratios were significantly less affected by flooding in SFF than in terra firme species. Although flood tolerance is critical for survival in flooded habitats, we hypothesize that responses to post-flooding events such as drought might be equally important in seasonal habitats. Therefore, we suggest that the ability to grow roots under anoxia might be critical in predicting success in inundated habitats that also experience a strong dry season. PMID- 12743795 TI - Analysis of zinc transporter, hZnT4 ( Slc30A4), gene expression in a mammary gland disorder leading to reduced zinc secretion into milk. AB - Zinc deficiency, causing impaired growth and development, may have a nutritional or genetic basis. We investigated two cases of inherited zinc deficiency found in breast-fed neonates, caused by low levels of zinc in the maternal milk. This condition is different from acrodermatitis enteropathica but has similarities to the "lethal milk" mouse, where low levels of zinc in the milk of lactating dams leads to zinc deficiency in pups. The mouse disorder has been attributed to a defect in the ZnT4 gene. Little is known about the expression of the human orthologue, hZnT4 (Slc30A4). Sequence analysis of cDNA, real-time PCR and Western blot analysis of hZnT4, carried out on control cells and cells from unrelated mothers of two infants with zinc deficiency, showed no differences. The hZnT4 gene was highly expressed in mouthwash buccal cells compared with lymphoblasts and fibroblasts. The hZnT4 protein did not co-localise with intracellular free zinc pools, suggesting that hZnT4 is not involved in transport of zinc into vesicles destined for secretion into milk. This observation, combined with phenotypic differences between the "lethal milk" mouse and the human disorder, suggests that the "lethal milk" mouse is not the corresponding model for the human zinc deficiency condition. PMID- 12743797 TI - A sero-epidemiological survey on the occurrence of opisthorchiid liver flukes in red foxes ( Vulpes vulpes) in Berlin, Germany. AB - Serum samples collected from red foxes in the city of Berlin between 1996 and 1999 were analysed for the presence of antibodies against Opisthorchis felineus and Metorchis bilis using an indirect ELISA. Out of 1,000 specimens, 30.6% and 46.5% reacted positively with specific O. felineus and M. bilis antigens, respectively. Seroprevalence in adult foxes was always higher than in juveniles. While no significant differences were observed in adult foxes throughout the period, in juvenile specimens seroprevalence declined from 1996 to 1997, then stayed at a comparable level in 1998 and increased in 1999. A varying availability of fresh cyprinid fish in different years seems to be the reason for changes in seroprevalence. By grouping the samples from juvenile foxes by season, antibodies against both Metorchis and Opisthorchis antigens started to appear between April and June, increased between July and September()and reached a level comparable to adult foxes in the October to December quarter. The lowest seroprevalence was found in Pankow, which is the district with the lowest share of the surface water. PMID- 12743798 TI - Molecular cloning and characterization of an SRCAP chromatin remodeling homologue in Toxoplasma gondii. AB - We have identified and mapped a gene in Toxoplasma gondii that encodes a homologue of SRCAP (Snf2-related CBP activator protein), a member of the SNF/SWI family of chromatin remodeling factors. The genomic locus (TgSRCAP) is present as a single copy and contains 16 introns. The predicted cDNA contains an open reading frame of 8,775 bp and encodes a protein of 2,924 amino acids. We have identified additional SRCAP-like sequences in Apicomplexa for comparison by screening genomic databases. An analysis of SRCAP homologues between species reveals signature features that may be indicative of SRCAP members. Expression of mRNA encoding TgSRCAP is upregulated when tachyzoite (invasive form) parasites are induced to differentiate into bradyzoites (encysted form) in vitro. Recombinant TgSRCAP protein is functionally equivalent to the human homologue, being capable of increasing transcription mediated by CREB. PMID- 12743799 TI - Malaria associated apoptosis is not significantly correlated with either parasitemia or the number of previous malaria attacks. AB - The occurrence and intensity of lymphocyte apoptosis in blood samples from 79 outclinic patients with uncomplicated Plasmodium falciparum or Plasmodium vivax malaria and 30 healthy individuals were investigated. No difference in apoptosis percentages was detected between healthy individuals and malaria patients when ex vivo lymphocytes were analyzed. However, significantly increased apoptosis levels were observed in lymphocytes from both P. falciparum- and P. vivax-infected patients when the cells were cultured for 24 h. CD4(+)and CD8(+) T cells were affected to a comparable extent in P.falciparum- and P.vivax-infected patients. However, when we compared apoptosis values in infected and non-infected individuals it appeared that CD4(+) T cells were more susceptible than CD8(+) T cells. A significant increase in the sIL-2R plasma levels was observed in malaria patients when compared with healthy individuals and a positive correlation was observed between sIL-2R levels and apoptosis rates in infected patients presenting increased rates of apoptosis. An increased expression of Fas antigen was recorded after stimulation with P. falciparum antigen or anti-CD3 monoclonal antibody. These data show that a consistent proportion of the lymphocyte population dies by apoptosis during a malaria infection and that a period of time is necessary before in vivo activated cells can express the apoptotic process in vitro. PMID- 12743800 TI - A new method for the experimental production of necrotic enteritis and its use for studies on the relationships between necrotic enteritis, coccidiosis and anticoccidial vaccination of chickens. AB - A new method for the experimental production of necrotic enteritis in chickens is described. The main features are the use of a diet high in wheat and fish meal content; oral administration of a non-lethal inoculum of the coccidium Eimeria maxima followed 6 days later by the bacterium Clostridium perfringens type A per cloaca, so that the bacterial inoculum is deposited at the time and place when and where the intestinal coccidial lesions are maximal; grading of coccidial and clostridial lesions in individual birds sampled during the 14 days following the coccidial infection. The new method was used to examine the relationship between clostridial and coccidial infections. Frank coccidiosis, caused by virulent E. maxima, exacerbated the lesions of necrotic enteritis and other clinical effects due to a subsequent challenge with virulent C. perfringens type A. Immunization with a live, pentavalent, attenuated anticoccidial vaccine (Paracox-5) protected against a severe challenge with heterologous E. maxima. Furthermore, vaccination with Paracox-5, by virtue of its protection against clinical coccidiosis due to the E. maxima challenge, indirectly protected birds against a subsequent challenge with virulent C. perfringens. The results are reconciled with previous field observations on concomitant coccidiosis and necrotic enteritis in chicken flocks. PMID- 12743801 TI - Characterization and cloning of metallo-proteinase in the excretory/secretory products of the infective-stage larva of Trichinella spiralis. AB - Inhibitor sensitivity assays using azocaesin and FTC-caesin as substrates showed that the excretory/secretory (E/S) products of the infective-stage larvae of Trichinella spiralis contained serine, metallo-, cysteine and aspartic proteinases. The activity of the metallo-proteinase was zinc ion dependent (within a range of ZnSO(4) concentrations). Gelatin-substrate gel electrophoresis revealed two bands of molecular mass 48 and 58 kDa which were sensitive to the metallo-proteinase inhibitor EDTA. The former peptide was probably a cleavage product of the latter. The authenticity of the 58 kDa metallo-proteinase as an E/S product was confirmed by immunoprecipitation. Using PCR and RACE reactions, a complete nucleotide sequence of the metallo-proteinase gene was obtained. It comprised 2,223 bp with an open reading frame encoding 604 amino acid residues. The 3' untranslated region consisted of 352 bp, including a polyadenylation signal AATAA. A consensus catalytic zinc-binding motif was present. The conserved domains suggest that the cloned metallo-proteinase belongs to the astacin family and occurs as a single copy gene with 11 introns and 10 exons. Cluster analysis showed that the sequence of the metallo-proteinase gene of T. spiralis resembles those of Caenorhabdites elegans and Strongyloides stercoralis. PMID- 12743803 TI - Hematological effects of Blastocystis hominis infection in male foreign workers in Taiwan. AB - Blastocystis hominis found in stool specimens has been the most frequently identified parasite among foreign workers from Southeast Asia in Taiwan since 1992. The prevalence of B. hominis was 14.1% in this study. In their quarantine physical examinations, 121 male Thai workers were examined hematologically and screened for stool parasites using the merthiolate-iodine-formaldehyde concentration method. Hematological values were compared in workers with and without a B. hominis infection. Multiple regressions were used to adjust for age. Those infected with any parasite other than B. hominis were excluded from further analysis. The workers infected with B. hominis had a lower leukocyte count (6.5+/ 0.4 X 10(3)/microl) than those who were not (7.4+/-0.2 X 10(3)/microl). This was mainly caused by a reduced neutrophil count (3.2+/-0.4 vs 4.2+/-0.2 X 10(3)/microl). Hemoglobin (13.9+/-0.3 vs 14.5+/-0.1 g/dl) and hematocrit (41.4+/ 0.6 vs 42.9+/-0.2%) were also reduced in B. hominis-positive workers. PMID- 12743802 TI - An aspartate aminotransferase of Wolbachia endobacteria from Onchocerca volvulus is recognized by IgG1 antibodies from residents of endemic areas. AB - Wolbachia are intracellular alpha-proteobacteria, closely related to Rickettsia, that infect various arthropods and filarial parasites. In the present study, the cDNA encoding the aspartate aminotransferase (AspAT) of Wolbachia from the human pathogenic filarial parasite Onchocerca volvulus (Ov-WolAspAT) was identified. At the amino acid level, the identity of the Ov-WolAspAT was 56% to Rickettsia prowazekii AspAT and 54% to the AspAT of the nitrogen-fixing bacterium Sinorhizobium meliloti, but the highest degree of identity was found to the putative AspAT of Wolbachia from Brugia malayi and Drosophila melanogaster (85%). All of these bacterial AspATs are members of the AspAT subclass Ib. A 35 kDa fragment of the Ov-WolAspAT was expressed in Escherichia coli, and immunolocalization using polyclonal antibodies against this antigen revealed that Ov-WolAspAT is present in a considerable proportion of the Wolbachia from O. volvulus, as well as in the endobacteria of several other filarial parasites. Western blot analysis using recombinant Ov-WolAspAT as antigen showed that IgG1 antibodies were present in 70 (51%) individuals living in areas endemic for O. volvulus, B. malayi or Wuchereria bancrofti and no IgG4 or IgE antibodies were found. Among 40 sera of persons from Uganda and Liberia who were putatively not infected with human filarial parasites, 11 (28%) individuals presented IgG1 antibodies, while none of the 33 sera from healthy Europeans and none of the 14 sera from patients with proven Rickettsia or Brucella infections reacted with the antigen. These results also show that an intracellular protein of Wolbachia endobacteria (WolAspAT) acts as antigen in human filariasis. PMID- 12743804 TI - Endocytobiont KC5/2 induces transformation into sol-like cytoplasm of its host Acanthamoeba sp. as substrate for its own development. AB - New investigations of a novel, recently described, non-cultivable endocytobiont of Acanthamoeba sp. reveal at least three hitherto unobserved developmental stages which shed some light on the nature of this peculiar organism. The development of the endocytobiont is closely connected with conspicuous changes in the host amoeba, inducing the transformation from gel to sol-like cytoplasm which bulges like a balloon inside the host cell. Young and transitory developmental stages were found within the homogenous, sol-like cytoplasm. The infectious stages, with their voluminous cell wall and a conspicuous ostiole, could be observed within all parts of the cytoplasm with the exception of the nucleus. It is a remarkable adaptation for this parasite to be able to induce this gel-sol transformation in order to facilitate its own development. The fate of the heavily infected host amoebae is death by rupture or lysis after being overcrowded with parasites. As no structures could be observed within the endoparasites that were comparable to other bacteria, the real nature and taxonomic position of these peculiar organisms remain obscure. PMID- 12743805 TI - Development of Trichostrongylus colubriformis and Trichostrongylus vitrinus, parasites of ruminants in the rabbit and comparison with Trichostrongylus retortaeformis. AB - The parasitic phase of development of both Trichostrongylus colubriformis and Trichostrongylus vitrinus, parasites of ruminants, was studied in detail in the rabbit. In T. colubriformis, the third moult appeared by 4 days after infection (DAI) and the last moult occurred between 10 and 11 DAI. In T. vitrinus, the third moult occurred between 8 and 11 DAI and the last one between 12 and 15 DAI. The prepatent period lasted 16-17 days for T. colubriformis and 20 days for T. vitrinus. The chronology of the life cycles and the distribution of the parasites along the small intestine for various Trichostrongylus spp. from lagomorphs and ruminants in the natural host or in the experimental host were compared. All of these biological parameters indicated a lower level of adaptation of T. vitrinus compared to the other species of Trichostrongylus. The results are fully compatible with the evolutionary scheme based on morphological analyses. PMID- 12743806 TI - A comparison of the structure of helminth communities in the woodmouse, Apodemus sylvaticus, on islands of the western Mediterranean and continental Europe. AB - We investigated the pattern of helminth species diversity in woodmouse, Apodemus sylvaticus, on western Mediterranean islands. We first performed a survey of the helminth fauna of A. sylvaticus in Sicily. Despite the small sampling effort, parasite species richness in Sicily is large in comparison with parasite species richness on other Mediterranean islands. We tested the nestedness of helminth parasite species from a number of Mediterranean localities using data compiled from epidemiological surveys of the helminth species of A. sylvaticus. We showed a nested pattern for woodmouse helminth species on western Mediterranean islands which suggests that the distribution of parasites on these islands is not the result of a random process. Properties of helminth parasites such as taxonomic group or life cycle and properties of the host population localities such as area size or the stability of the environment (estimated by mammal species diversity) contribute to the nestedness. PMID- 12743809 TI - First record of a Mermithidae (Nematoda) from the meloid beetle Meloe violaceus Marsham, 1802 (Coleoptera: Meloidae). AB - A new record of nematode parasitism of meloid beetles is reported and all earlier records are summarised. Rates of parasitism could be influenced by the toxic compound cantharidin that these beetles possess. PMID- 12743807 TI - Arsenicals (melarsoprol), pentamidine and suramin in the treatment of human African trypanosomiasis. AB - Human African trypanosomiasis (HAT), otherwise known as sleeping sickness, has remained a disease with no effective treatment. Recent progress in HAT research suggests that a vaccine against the disease is far from being successful. Also the emergence of drug-resistant trypanosomes makes further work in this area imperative. So far the treatment for the early stage of HAT involves the drugs pentamidine and suramin which have been very successful. In the second stage of the disease, during which the trypanosomes reside in the cerebrospinal fluid (CSF), treatment is dependent exclusively on the arsenical compound melarsoprol. This is largely due to the inability to find compounds that can cross the blood brain barrier and kill the CSF-residing trypanosomes. This review summarises our current understanding on the treatment of HAT. PMID- 12743808 TI - First occurrence of Mesocestoides sp. in a bird, the red-legged partridge, Alectoris rufa, in Spain. AB - Larvae of Mesocestoides sp. were found in the celomic cavity of one red-legged partridge (Alectoris rufa) from Azuara (Aragon, north-eastern Spain). This is the first report of this larval cestode occurring in birds from Spain. None of the other 36 partridges from this area nor 150 partridges from other Spanish localities were affected. The red-legged partridge does not seem to be an important link in the Mesocestoides sp. life-cycle. PMID- 12743810 TI - Cloning, sequencing and recombinant expression of the open reading frame encoding a novel member of the Sarcocystis muris (Apicomplexa) microneme lectin family. AB - Micronemes are characteristic secretory organelles located within the apical cell region of apicomplexan parasites. The protein contents are exocytosed during an early phase of host cell invasion and contribute to parasite motility and the invasion of target cells. We report here on the cloning and heterologous expression of a novel member of the Sarcocystis muris microneme lectin family. The deduced amino acid sequence is in total agreement with that obtained after sequencing the native protein and is characterized by two copies of the apple domain motif. The recombinant polypeptide is expressed in a biologically active conformation as demonstrated by its galactose binding properties. PMID- 12743811 TI - High susceptibility of nullizygous p53 knockout mice to colorectal tumor induction by 1,2-dimethylhydrazine. AB - PURPOSE: The susceptibility of male p53 nullizygote (-/-), heterozygote (+/-), and wild-type (+/+) mice to 1,2-dimethylhydrazine (DMH) induction of colon carcinogenesis was investigated. METHODS: In a preliminary short-term experiment, male mice of three genotypes were given s.c. of 20 mg/kg DMH once weekly for 5 weeks. In a medium-term experiment, mice were given weekly s.c. of DMH for 15 weeks. In a long-term experiment, male p53 (+/-) and (+/+) mice were given weekly injections of DMH for 15 weeks, and killed at week 30. RESULTS: In the medium term experiment, carcinomas were observed in 70% of p53 (-/-) mice, although there were no carcinomas in p53 (+/+) and (+/-) mice. In the long-term experiment, there was no significant difference in incidences of adenomas and carcinomas between p53 (+/+) and (+/-) mice. PCR-single strand conformation polymorphism analysis of exons 5-8 of p53 gene revealed four mutations in one focal atypia, one adenoma, and two carcinomas, out of 56 colonic proliferative lesions in the medium- and long-term experiments. CONCLUSIONS: These results suggest that p53 might not be a direct target of DMH but complete loss of p53 might elevate susceptibility to DMH-induced colorectal carcinogenesis. PMID- 12743812 TI - Major components of the insulin-like growth factor axis are expressed early in chicken embryogenesis, with IGF binding protein ( IGFBP) -5 expression subject to regulation by Sonic Hedgehog. AB - Using whole-mount in situ hybridisation techniques, we have examined the expression of major components of the insulin-like growth factor (IGF) axis in early development of the chicken embryo, including both IGF-I and -II, the type 1 IGF receptor ( IGFR), and two of the IGF binding proteins, ( IGFBP) -2 and -5. We report that these genes fall into two distinct groups with respect to expression pattern, with IGFBP-2 displaying broad overlap of mRNA expression with IGFR and IGF-I during early development, whereas the expression profile of IGFBP-5 most closely resembled that of IGF-II. Comparison between different stages revealed IGFBP-2 mRNA was detected as early as stage 3, whereas IGFBP-5 was first seen at stage 4. In addition, we detected expression domains of IGFBP-5, and to a lesser extent IGFBP-2, which did not overlap with either IGFR or IGF expression patterns. This could indicate IGF independent actions of the IGFBPs during early embryonic development. A striking observation concerning the expression profiles of both IGF-II and IGFBP-5 at early stages of chick embryogenesis is that both these genes are expressed asymmetrically in a pattern similar to that of Sonic Hedgehog (Shh). Furthermore, using cyclopamine, we have demonstrated that IGFBP-5 expression in the early embryo is regulated by Shh. Taken together, these results describe an important role for the IGF system in the very early stages of the developing chicken embryo, and imply that IGFBP-2 and -5 are fundamental developmental factors, with the latter involved in Shh signalling pathways. PMID- 12743813 TI - Proliferative activities of tumor stromal cells play important roles in tumor thickness and progression of T3 ulcerative-type colorectal cancer. AB - The central depressed area thickness (tumor thickness) of colorectal cancers is an important prognostic parameter for colorectal cancer patients. We examined whether the proliferative activities of tumor and stromal components play important roles in the increase of tumor thickness and the progression of colorectal cancers. Colorectal cancers were classified into thin and thick groups according to tumor thickness. The proliferative activities of fibroblasts and endothelial cells were immunohistochemically evaluated in 157 T3 ulcerative-type colorectal cancers by CD31/MIB-1 (anti-Ki-67 antigen) double staining. The MIB-1 labeling index was estimated as the percentage of fibroblasts with positive nuclei. The CD31-positive microvessels lined by MIB-1-positive endothelial cells were assessed. The proliferative microvessel index was defined as the percentage of proliferative microvessels relative to all microvessels. The fibroblast MIB-1 labeling index was the only parameter significantly associated with tumor thickness ( P=0.049). High fibroblast MIB-1 labeling indices showed significant correlation with tumor recurrence in the thin group ( P=0.020). High proliferative microvessel index was a significant parameter of tumor recurrence in the thick group ( P=0.003) in multivariate analyses. This study strongly suggests that proliferative activities of stromal components are useful parameters of tumor biology and of prognosis for T3 ulcerative-type colorectal cancer patients. PMID- 12743814 TI - Misalignment of lung vessels: diagnostic role of conventional histology and immunohistochemistry. AB - Misalignment of lung vessels represents a rare congenital anomaly that may cause respiratory failure in the newborn. It is characterized by abnormal position of pulmonary veins and venules that lie adjacent to arteries and bronchi, and it is usually associated with a decreased number of alveolar capillaries (i.e., alveolar capillary dysplasia), although these two conditions have been separately described. Awareness of this anomaly is required by pathologists because it can be easily overlooked on lung biopsy or autopsy, and because definite diagnosis relies on histology. We report the case of a newborn male baby who developed respiratory distress 18 h after an uncomplicated delivery. The patient died on the 7th day, after high frequency oscillatory ventilation, nitric oxide inhalation and extracorporeal membrane oxygenation were unsuccessful. On autopsy, histology and immunohistochemistry demonstrated diffuse changes, fulfilling diagnostic criteria of misalignment of lung vessels and of alveolar capillary dysplasia in both lungs, with muscularization of very peripheral pulmonary arteries and a prominent interstitial and periadventitial fibrosis. Diffuse distribution of vessel misalignment could explain the rapid onset of respiratory failure, and the presence of diffuse fibrosis might have contributed to irreversible respiratory dysfunction by impairment of lung parenchyma extensibility. PMID- 12743816 TI - Iliopsoas muscle bleeding as a complication of septic disseminated intravascular coagulation. PMID- 12743815 TI - Expression of drug resistance related proteins in sarcomas of the pulmonary artery and poorly differentiated leiomyosarcomas of other origin. AB - Sarcomas are known to develop resistance to current chemotherapeutic strategies, displaying a multidrug-resistant phenotype. Mechanisms involved in drug resistance include reduced cellular drug accumulation, drug detoxification as well as alterations in drug target specificity. In seven sarcomas of the pulmonary artery (SPA) and ten leiomyosarcomas of other origin, we studied the immunohistochemical expression of P-glycoprotein (P-gp), multidrug-resistance protein (MRP), lung resistance protein (LRP), metallothionein (MT) and topoisomerase IIalpha. Upregulation was found in tumour cells for P-gp but not for MRP in SPA and other leiomyosarcomas. Topoisomerase IIalpha was expressed at high levels in tissue of primary tumours as well as recurrent tumours. Both P-gp and topoisomerase IIalpha were present in numerous tumour-associated vessels. LRP was expressed at high levels in SPA but to a lesser extent in the other leiomyosarcomas. MT was expressed at low levels but was markedly present at the border of necrosis. The overall survival and the relapse-free survival did not correlate with the expression of these factors. There was no significant relationship between treated and non-treated patients with respect to the expression of the examined molecules. P-gp, but not MRP, may play a role in the development of drug resistance. P-gp, LRP and topoisomerase IIalpha contribute to drug resistance through expression in tumour-associated vessels. Unique high levels of topisomerase IIalpha reflect the high proliferation rate of these tumours. MT seems to serve as a detoxifying agent of metabolites at the border of necrosis. Our findings underline the fact that multiple factors contribute to chemoresistance and that examination of a spectrum of relevant molecules is probably necessary to plan the best therapy. PMID- 12743817 TI - Invasive lobular carcinoma arising in phyllodes tumor of the breast. PMID- 12743818 TI - CD31 staining in epithelioid sarcoma. AB - We report an unusual case of epithelioid sarcoma. The tumour occurred in the finger of a 27-year-old female. The clinical history, histology and the electron microscopy of the lesion were typical for epithelioid sarcoma. However, immunohistochemical analysis showed strong membranous CD31 staining, a finding hitherto not described. All other robust vascular markers, including factor-VIII related antigen (FVIIIrag) were negative. The findings were compared with the available literature data, leading us to conclude that there is insufficient evidence for endothelial derivation of epithelioid sarcoma, but in the differential diagnosis with vascular tumours CD31 may stain and to rule out angiosarcoma FVIIIrag is a useful antibody. PMID- 12743819 TI - A genomewide survey of developmentally relevant genes in Ciona intestinalis. IV. Genes for HMG transcriptional regulators, bZip and GATA/Gli/Zic/Snail. AB - Many kinds of transcription factors and regulators play key roles in a variety of developmental processes. In the present survey, genes encoding proteins with conserved HMG-box, bZip domains, and some types of zinc finger motifs were surveyed in the completely sequenced genome of Ciona intestinalis. In the present analysis, 21 HMG-box-containing genes and 26 bZip genes were identified as well as four small groups of zinc finger genes in the Ciona genome. The results also showed that a less redundant set of genes is present in the Ciona genome compared with vertebrate genomes. In addition, cDNA clones for almost all genes identified have been cloned and distributed as a " Ciona intestinalis Gene Collection Release I". The present comprehensive analysis therefore provides a means to study the role of these transcription factors in developmental processes of basal chordates. PMID- 12743820 TI - A genomewide survey of developmentally relevant genes in Ciona intestinalis. III. Genes for Fox, ETS, nuclear receptors and NFkappaB. AB - A survey against the draft genome sequence and the cDNA/EST database of Ciona intestinalis identified a number of genes encoding transcription factors regulating a variety of processes including development. In the present study, we describe almost complete sets of genes for Fox, ETS-domain transcription factors, nuclear receptors, and NFkappaB as well as other factors regulating NFkappaB activity, with their phylogenetic nature. Vertebrate Fox transcription factors are currently delineated into 17 subfamilies: FoxA to FoxQ. The present survey yielded 29 genes of this family in the Ciona genome, 24 of which were Ciona orthologues of known Fox genes. In addition, we found 15 ETS genes, 17 nuclear receptor genes, and several NFkappaB signaling pathway genes in the Ciona genome. The number of Ciona genes in each family is much smaller than that of vertebrates, which represents a simplified feature of the ascidian genome. For example, humans have two NFkappaB genes, three Rel genes, and five NFAT genes, while Ciona has one gene for each family. The Ciona genome also contains smaller numbers of genes for the NFkappaB regulatory system, i.e. after the split of ascidians/vertebrates, vertebrates evolved a more complex NFkappaB system. The present results therefore provide molecular information for the investigation of complex developmental processes, and an insight into chordate evolution. PMID- 12743821 TI - A genomewide survey of developmentally relevant genes in Ciona intestinalis. VIII. Genes for PI3K signaling and cell cycle. AB - Cell growth and cell divisions are two fundamental biological processes for cells in multi-cellular organisms. The molecules involved in these biological processes are highly conserved within eukaryotes, including plants and unicellular organisms such as yeast. However, some regulatory molecules seem to be innovated during animal evolution. Therefore, to understand how the ubiquitous systems have evolved or have been conserved, we examined genes for the phosphoinositide 3 kinase (PI3K) pathway that is important for cell growth, and genes for cell cycle regulation in the genome of Ciona intestinalis. It was found that the Ciona intestinalis genome contains all the essential constituents of the PI3K pathway. In addition, the class IB PI3K catalytic and regulatory subunits, which had not previously been known in animals other than mammals, were found in the Ciona genome. Similarly, all essential cyclins and CDKs were found in the Ciona genome, while cyclin G and cyclin L were likely to be independently lost in the ascidian lineage, which may be dispensable for the cell cycle. Cyclin F, which was previously known only in vertebrates, was not found in the Ciona genome. Therefore, this gene was probably innovated during the evolution of vertebrates to be involved in vertebrate-specific cell cycle regulation. Since Ciona is regarded as one of the most primitive extant chordates, the present analysis gives us an insight into how these fundamental biological genes are evolved or are conserved during chordate evolution. PMID- 12743822 TI - Loss of CLE40, a protein functionally equivalent to the stem cell restricting signal CLV3, enhances root waving in Arabidopsis. AB - Continuous growth and development of plants is controlled by meristems that harbour stem cell pools. Division of stem cells and differentiation of their progeny are coordinated by intercellular signaling. In Arabidopsis, stem cells in shoot and floral meristems secrete CLAVATA3, a member of the CLE protein family that activates the CLV1/CLV2 receptor complex in underlying cells to restrict the size of the stem cell population. We found that CLE40 encodes a potentially secreted protein that is distantly related to CLV3. While CLV3 transcripts are confined to stem cells of the shoot system, CLE40 is expressed at low levels in all tissues, including roots. Misexpression and promoter swap experiments show that CLE40 can fully substitute for CLV3 to activate CLV signalling in the shoot, indicating that CLV3 and CLE40 are functionally equivalent proteins that differ mainly in their expression patterns. Analysis of cle40 mutants shows that wild type expression levels of CLE40 are insufficient to contribute to CLV signalling. High level expression of CLV3 or CLE40 results in a premature loss of root meristem activity, indicating that activation of a CLV-like signaling pathway may restrict cell fate also in roots. The cellular organization of cle40 root meristems is normal, but mutant roots grow in a strongly waving pattern, suggesting a role for CLE40 in a signaling pathway that controls movement of the root tip. PMID- 12743823 TI - The role of GlsA in the evolution of asymmetric cell division in the green alga Volvox carteri. AB - Volvox carteri, a green alga in the order Volvocales, contains two completely differentiated cell types, small motile somatic cells and large reproductive cells called gonidia, that are set apart from each other during embryogenesis by a series of visibly asymmetric cell divisions. Mutational analysis has revealed a class of genes (gonidialess, gls) that are required specifically for asymmetric divisions in V. carteri, but that are dispensable for symmetric divisions. Previously we cloned one of these genes, glsA, and showed that it encodes a chaperone-like protein (GlsA) that has close orthologs in a diverse set of eukaryotes, ranging from fungi to vertebrates and higher plants. In the present study we set out to explore the role of glsA in the evolution of asymmetric division in the volvocine algae by cloning and characterizing a glsA ortholog from one of the simplest members of the group, Chlamydomonas reinhardtii, which does not undergo asymmetric divisions. This ortholog (which we have named gar1, for glsA related) is predicted to encode a protein that is 70% identical to GlsA overall, and that is most closely related to GlsA in the same domains that are most highly conserved between GlsA and its other known orthologs. We report that a gar1 transgene fully complements the glsA mutation in V. carteri, a result that suggests that asymmetric division probably arose through the modification of a gene whose product interacts with GlsA, but not through a modification of glsA itself. PMID- 12743824 TI - Triacylglycerols determine the unusual storage physiology of Cuphea seed. AB - Many species within the genus Cuphea (Lythraceae) produce seed with high levels of medium-chain fatty acids. Seeds of some Cuphea species lose viability when placed into storage at -18 degrees C. These species tolerate significant drying to 0.05 g/g and may, therefore, be intermediate in their storage characteristics. The thermal properties of seed lipids were observed using differential scanning calorimetry. Species with peak lipid melting temperatures >/=27 degrees C were found to be sensitive to -18 degrees C exposure while those with melting temperatures <27 degrees C were able to tolerate low-temperature exposure. This relationship was determined by the triacylglycerol composition of the individual species. Sensitive species have high concentrations of lauric acid (C(12)) and/or myristic acid (C(14)). Species with high concentrations of capric (C(8)) or caprylic acid (C(10)) or with high concentrations of unsaturated fatty acids tolerate low temperature exposure. Potential damage caused by low temperature exposure can be avoided by exposing seeds to a brief heat pulse of 45 degrees C to melt solidified lipids prior to imbibition. The relationship between the behavior of triacylglycerols in vivo, seed storage behavior and sensitivity to imbibitional damage is previously unreported and may apply to other species with physiologies that make them difficult to store. PMID- 12743825 TI - A 10-kDa class-CI sHsp protects E. coli from oxidative and high-temperature stress. AB - We report on a new cDNA clone (Qshsp10.4-CI) of a Quercus suber L. class-CI small heat-shock protein (sHsp) obtained from cork (phellem), a highly oxidatively stressed plant tissue. The deduced gene product lacks the C-terminal extension and the consensus I region of the alpha-crystallin domain, being the most C terminally truncated sHsp reported to date. In an attempt to prove that a protective function is possible for such a truncated sHsp, we overexpressed in Escherichia coli three recombinant sHsp-CIs, one (rQsHsp10.4-CI) showing the same truncation as Qshsp10.4-CI, a second (rN49) lacking the whole alpha-crystallin domain, and a third (rN153) consisting of a full-length sHsp-CI. The overexpression of rN153 and, remarkably, rQsHsp10.4-CI but not rN49 enhanced cell viability under high temperature and, interestingly, under oxidative stress. These results show that the C-terminal extension and the consensus I region of the alpha-crystallin domain are dispensable, but amino acids 1-41 of the alpha crystallin domain (including the consensus II region) are essential for the protective activity of sHsp-CIs. On the other hand, two-dimensional immunodetection patterns showed accumulation of ca. 10-kDa sHsp-CI immunorelated polypeptides in cork and other oxidatively stressed tissues but not in control and heat-stressed tissues. We discuss the possible role of highly truncated sHsps in relation to oxidative stress. PMID- 12743827 TI - Glycodelin and amniotic fluid transferrin as inhibitors of E-selectin-mediated cell adhesion. AB - Human amniotic fluid contains a variety of glycoproteins. Several of these substances have been shown to exert immunomodulatory effects. Glycodelin, previously known as placental protein 14, is one of these glycoproteins. It has a unique carbohydrate configuration, consistent with fucosylated LacdiNAc structures that are very unusual for mammals. Oligosaccharides with fucosylated LacdiNAc antennae have previously been shown to block selectin-mediated cell adhesion. Another glycoprotein, human transferrin, is also present in amniotic fluid in relatively high concentrations. This transferrin shows a different glycosylation compared with serum transferrin. Amniotic fluid transferrin carries sialylated Lewis X antigens. Glycodelin and transferrin were isolated from amniotic fluid and for comparison from serum of pregnant women by chromatographic methods. The purified proteins were used as ligands to block E-selectin-mediated HepG2 cell adhesion. Two types of binding assays with distinct receptor accommodations (immobilised E-selectin and activated HUVECs) were used to quantify inhibition efficiencies of the different proteins. We found that glycodelin is a strong inhibitor with a 10(3)-fold potency compared to the monovalent tetrasaccharide sialyl Lewis X whereas the potency of transferrin is rather low. PMID- 12743826 TI - Characterisation of the progression of azaserine-induced rat pancreatic adenocarcinoma by proliferative cell nuclear antigen, basement membrane laminin and trypsinogen immunohistochemistry. AB - The progression of azaserine-induced rat pancreatic adenocarcinoma (AC) was characterised using quantitative and semiquantitative immunohistochemistry for proliferating cell nuclear antigen (PCNA), basement membrane laminin (BML) and trypsinogen (TG). Samples were taken 5-20 months after initiation. High PCNA labelling indices (PCNA LIs) were measured 5 months after the induction of atypical acinar cell nodules (AACNs), which decreased later and stagnated until a further decline in the month 10 adenomas. Then a second premalignant proliferative wave was observed (month 13) within the adenoma stage. Later, in month 20 differentiated ACs PCNA LIs fell to the host tissue level but were found highest in the month 20 anaplastic ACs indicating a switch to malignant proliferation. Month 20 invasive ACs showed a number of separate proliferative foci. In early AACNs, BML decreased and remained low till the local maximum in the month 13 adenoma. Invasive ACs did not express BML. Month 5 AACN and differentiated AC were TG deficient but anaplastic AC regained its TG expression. However invasive AC was again TG negative. These results are discussed in combination with our previous data on progressional changes of autophagic capacity and microvessel densities. PMID- 12743828 TI - Effects of Vitreoscilla hemoglobin on the 2,4-dinitrotoluene (2,4-DNT) dioxygenase activity of Burkholderia and on 2,4-DNT degradation in two-phase bioreactors. AB - Expression of vgb, encoding Vitreoscilla hemoglobin (VHb), in Burkholderia strain YV1 was previously shown to improve cell growth and enhance 2,4-dinitrotoluene (2,4-DNT) degradation compared with control strain DNT, especially under hypoxic conditions. In the work reported here, the ratio of 2,4-DNT degraded to oxygen uptake was approximately 5-fold larger for strain YV1 than for strain DNT. The addition of purified VHb to cytosolic fractions of strain DNT increased 2,4-DNT degradation 1.5-fold, compared with 1.1-fold for control bovine Hb, but increased the 2,4-DNT degradation 2.7-fold when added to partially purified 2,4-DNT dioxygenase, compared with 1.3-fold for bovine Hb. This suggests a direct transfer of oxygen from VHb to the oxygenase. In a bioreactor at high 2,4-DNT concentration (using 100 ml oleyl alcohol containing 2 g 2,4-DNT as the second phase) with 1.5 l culture, both strains could remove 0.8 g 2,4-DNT by 120 h; and, under the same conditions in a fed-batch reactor, the degradation increased to 1 g for strain YV1 but not for strain DNT. PMID- 12743830 TI - Visible trends in functional genomics. PMID- 12743831 TI - Peripartum tuberculosis as a form of immunorestitution disease. AB - The aim of this study was to assess the clinical spectrum of peripartum tuberculosis from the perspective of immunorestitution disease. Of 29 patients with peripartum tuberculosis, 27 (93.1%) had extrapulmonary tuberculosis, 20 (69%) of whom were affected in the central nervous system. Twenty-two (75.9%) patients had no clinical features suggestive of tuberculosis during pregnancy. The median time from delivery to the onset of immunorestitution was 4 days, but treatment with anti-tuberculous therapy was delayed for a median time of 27 days after the onset of symptoms. Despite therapy, 11 (38%) patients died and 4 (13.8%) had residual functional deficits. Peripartum tuberculosis is an important differential diagnosis of postpartum fever (of unknown origin) without localized signs. PMID- 12743832 TI - High prevalence of superantigens associated with the egc locus in Staphylococcus aureus isolates from patients with atopic eczema. AB - The present study was aimed at identifying a possible correlation between disease severity and colonization with superantigen-producing Staphylococcus aureus strains in patients with atopic eczema. To this end, Staphylococcus aureus strains from 91 patients with atopic eczema were screened for various staphylococcal superantigens such as SEA, SEB, SEC, SED, TSST1, the recently described enterotoxin gene cluster egc (which encodes the enterotoxins SEG, SEI, SEK, SEM, and SEO), and the see, seh, and sej loci. Swabs were taken from seven different sites in each patient. The rate of colonization with Staphylococcus aureus was 87.9%. Of those patients colonized, 35% were colonized with more than one different strain. Of the 120 genetically different strains investigated, the egc locus was found in 48.3% and the sej locus in 7.5%. The see and seh loci were not found in any strain. The presence of the classical superantigens SEA-SED or TSST1 was found in 38.3%. Overall, 71.3% of the Staphylococcus aureus-positive patients harbored at least one superantigen-producing strain on their skin. There was no difference in the prevalence of superantigens between atopic eczema patients and healthy volunteers. Moreover, there was no difference in the extent of disease expression between patients colonized by superantigen-positive Staphylococcus aureus strains and those with superantigen-negative strains as measured by the SCORAD system. However, patients colonized with Staphylococcus aureus had a significantly higher SCORAD score than those not colonized. PMID- 12743833 TI - Distribution and phylogenetic diversity of the subsurface microbial community in a Japanese epithermal gold mine. AB - Distribution and phylogenetic diversity of microbial communities in hot, deep underground environments in the Hishikari epithermal gold mine, southern part of Kyushu, Japan, were evaluated using molecular phylogenetic analyses. Samples included drilled cores such as andesitic volcanic rock (0.95-1.78 Ma) and the oceanic sedimentary basement rock of Shimanto-Supergroup (100 Ma), as well as geothermal hot aquifer waters directly collected from two different sites: AW site (71.5 degrees C, pH 6.19) and XW-site (85.0 degrees C, pH 6.80) at a depth of 350 mbls (meters below land surface). Based on PCR-amplified 16S rRNA gene clone analysis, the microbial communities in the drilled cores and the hot aquifer water from the XW-site consisted largely of the 16S rRNA gene sequences, closely related to the sequences often found in marine environments, while the aquifer water from the AW-site contained 16S rRNA gene sequences representing members of Aquificales, thermophilic methanotrophs within the gamma-subdivision of the Proteobacteria and uncultivated strains within the beta-subdivision of Proteobacteria. The cultivable microbial community detected by enrichment cultivation analysis largely matched that detected by the culture-independent molecular analysis. PMID- 12743834 TI - Desulfurobacterium crinifex sp. nov., a novel thermophilic, pinkish-streamer forming, chemolithoautotrophic bacterium isolated from a Juan de Fuca Ridge hydrothermal vent and amendment of the genus Desulfurobacterium. AB - A novel thermophilic, chemolithoautotrophic bacterium, designated as NE1206(T), was isolated from a Juan de Fuca Ridge hydrothermal vent sample (tubes of the annelid polychaete Paralvinella sulfincola attached to small pieces of hydrothermal chimney). The cells were rod-shaped (1.2-3.5 x 0.4-0.7 microm), occurring as single motile rods or forming macroscopic aggregates visible as pinkish to brownish streamers. The new isolate was anaerobic. It grew between 50 and 70 degrees C (optimum 60-65 degrees C; doubling time approximately 1 h 15 min at 60 degrees C), between pH 5.0 and 7.5 (optimum pH around 6.0-6.5) and at sea salts concentrations between 20 and 40 g l(-1 )(optimum 30 g l(-1)). Cells grew chemolithoautotrophically in an H(2)/CO(2) atmosphere (80/20, v/v; 200 kPa). Molecular hydrogen was the sole electron donor used by the strain. Nitrate and elemental sulfur served as electron acceptors, yielding ammonia and hydrogen sulfide, respectively (nitrate reduction supported higher growth rates than sulfur reduction). The G+C content of the genomic DNA was 36.7+/-0.8 mol%. Phylogenetic analyses of the 16S rRNA gene located the strain within the genus Desulfurobacterium. However, the novel isolate possesses physiological and biochemical characteristics that differ from the previously described species of this genus. We propose that the isolate represents a novel species, Desulfurobacterium crinifex sp. nov. The type strain is NE1206(T) (DSM 15218(T), CIP 107649(T)). An amendment of the genus Desulfurobacterium description is proposed, based on the phenotypic characteristics of the novel species. PMID- 12743835 TI - Comparative genomic analysis of solvent extrusion pumps in Pseudomonas strains exhibiting different degrees of solvent tolerance. AB - Organic solvents are inherently toxic for microorganisms. Their effects depend not only on the nature of the compound, but also on the intrinsic tolerance of the bacterial species and strains. Three efflux pumps belonging to the RND (resistance-nodulation-cell division) family of multidrug extrusion pumps are the main factor involved in the high intrinsic tolerance to toluene of Pseudomonas putida DOT-T1E. We have analyzed the tolerance to toluene shocks [0.1% and 0.3% (v/v)] of a number of strains belonging to different species of the genus Pseudomonas upon growth in the absence and in the presence of sublethal concentrations of toluene. The strains can be grouped in three categories: (1) highly resistant strains, in which almost 100% of the cells precultured in the presence of sublethal concentrations of toluene withstood a 0.3% (v/v) toluene shock, (2) moderately resistant strains, in which only a fraction (10(-4)-1) of the cells withstood a 0.1% (v/v) toluene shock, but fewer than 1 in 10(7) cells survived a sudden 0.3% (v/v) toluene shock regardless of the growth conditions, and (3) sensitive strains, in which regardless of the growth conditions fewer than 10(-5) cells survived a 0.1% (v/v) toluene shock. We also studied the number and type of efflux pumps in different strains in comparison with the P. putida DOT-T1E strain. PMID- 12743836 TI - Long-term clinical performance and longevity of gold alloy vs ceramic partial crowns. AB - Cast gold partial crowns (CGPC) are an accepted means of restoring posterior teeth. For aesthetic reasons, gold alloys are being increasingly substituted with ceramics. The aim of the present study was to investigate retrospectively the long-term clinical performance and survival of CGPC and compare the results to the ones already reported for ceramic partial crowns (CPC). The CGPC group consisted of 42 patients (24 male, 18 female) randomly sampled from a total of 106 patients with CGPC, with one restoration per patient. The CPC group consisted of 22 patients with a total of 42 restorations. Both types of restoration were done by one experienced dentist. Another two experienced dentists who were not involved in performing the restorations rated both kinds of partial crowns using the modified United State Public Health Service (USPHS) criteria [14]. The Median age of the CGPC was 57 months (range 3-157) and of the CPC and 63 months (range 24-72). Forty-one (98%) of the CGPC and 27 (64%) of the CPC were placed in molars, the rest in premolars. In each group, 40 (95%) restorations were still functioning without any necessity of replacement. Two teeth with CGPC, in situ for 4.5 and 11 years, respectively, had been extracted for periodontal reasons. Two CPC fractured and had to be replaced after 2 and 6.5 years in situ. The USPHS criteria results were similarly good for the gold and ceramic groups. Kaplan Meier analysis revealed survival probabilities of 72+/-21% and 96+/-4% after 13 and 7 years, respectively, for the CGPC. Survival of the CPC was 81+/-15% after 7 years. No statistically significant difference among survival functions of CGPC and CPC was found. From this data, it can be concluded that the longevity of CPC is not inferior to that of gold alloys. However, more long-term studies comparing the clinical performance and longevity of these two types of indirect restoration in the posterior region with larger numbers of restorations are desirable. PMID- 12743837 TI - Decrease in the total antioxidant activity of saliva in patients with periodontal diseases. AB - This study examined the role of free radical-induced tissue damage and the antioxidant defense mechanism of saliva in periodontal disease. Antioxidant activity of saliva was compared in 20 healthy individuals and 17 patients with periodontal diseases. We measured the scavenging capacity of saliva against free radicals generated in vitro by electrolysis, xanthine-xanthine oxidase, or stimulated polymorphonuclear leukocytes. Total protein content and total antioxidant activity of saliva were also determined. The results indicate that stimulated saliva of healthy individuals is significantly more effective (40-50%) than that of patients with periodontal diseases in scavenging a wide variety of free radicals generated in vitro. Under these conditions it appears that the total antioxidant activity of saliva is significantly decreased in these patients despite the fact that the levels of the three main antioxidants (uric acid, ascorbic acid, and albumin) are not significantly affected. We conclude that periodontal diseases are associated with an imbalance between oxidants and antioxidants in favor of the former due to both an increase in free radical production and a defect in the total antioxidant activity of saliva. PMID- 12743839 TI - [The maestro, the method and the memory]. PMID- 12743838 TI - Mycorrhizal promotion of host stomatal conductance in relation to irradiance and temperature. AB - Colonization of roots and soil by arbuscular mycorrhizal (AM) fungi sometimes promotes stomatal conductance (gs) of the host plant, but scientists have had difficulty predicting or manipulating the response. Our objective was to test whether the magnitude of AM influence on gs is related to environmental conditions: irradiance, air temperature or leaf temperature. Stomatal conductances of two groups of uncolonized sorghum plants were compared to gs of plants colonized by Glomus intraradices (Gi) or Gigaspora margarita (Gm) in 31 morning and afternoon periods under naturally varying greenhouse conditions. Stomatal conductance of Gi and Gm plants was often markedly higher than gs of similarly sized nonAM plants. AM promotion of gs was minimal at the lowest irradiances and lowest air and leaf temperatures, but was substantial at intermediate irradiance and temperatures. AM promotion was again low or absent at the highest irradiances and temperatures. Magnitude of AM promotion of gs was not a function of absolute gs. Promotion of gs by Gi and Gm was remarkably similar. Differing phosphorus fertilization did not affect gs. PMID- 12743840 TI - [Research in refractive surgery and quality of vision]. PMID- 12743841 TI - [Cataract surgery: viewpoints of some ophthalmologists]. PMID- 12743842 TI - [Comparative study of two portable tonometers: Tono-Pen XL and Perkins]. AB - PURPOSE: To evaluate the reproducibility of two portable tonometers and to compare the intraocular pressure (IOP) measurements obtained with both instruments taking into account the level of experience of the examiner with this technique. METHOD: Cross-sectional study. IOP was measured in 35 eyes of 35 normal subjects and 20 eyes of 20 ocular hypertensive patients with Perkins aplanation tonometer and Tono-Pen XL digital tonometer. Five measurements were made with Perkins (by an ophthalmologist) and 10 with Tono-Pen (5 by an ophthalmologist and 5 by a general practitioner). RESULTS: Considering the sample as a whole there was no statistically significant difference, and there was a significant correlation between the mean measurements obtained with Perkins tonometer and Tono-Pen and between the mean obtained by doctors with and without previous tonometric experience (p<0.001, ANOVA). Variability of Tono-Pen measurements is greater than the Perkins measurements and it is considerable in both cases (coefficient of variation 7-9%). Tono-Pen measurements tend to show a greater variability in hypertensive subjects, and tend to underestimate high IOP when compared with Perkins measurements. Tono-Pen identified IOP > or =21 mmHg with a sensitivity of 40% and a specificity of 94-97%. CONCLUSION: Tono-Pen allows measurement of IOP to personal with no tonometric experience. There is a strong correlation between measurements obtained with both tonometers. Tono-Pen shows a tendency to underestimate high IOP and offers low sensitivity in detecting IOP greater than 21 mmHg. PMID- 12743843 TI - [Estimation of the efficacy of deep sclerectomy in glaucoma surgery. One-year follow-up]. AB - PURPOSE: To evaluate the efficacy of deep sclerectomy in the surgical treatment of glaucoma one year after surgery. METHODS: 53 eyes of 43 patients (26 male, 27 female) with medically uncontrolled glaucoma of various types, were treated with deep sclerectomy surgery. RESULTS: The mean postoperative intraocular pressure (IOP) was 18.19 S.D. 5.22 mmHg., and the mean preoperative IOP was 26.66 S.D. 5.93 mmHg. We observed a statistically significant reduction in the IOP (p<0.05). Our success criteria were IOP lower than 21 mmHg with or without associated hypotensive medications, no advance in glaucomatous visual field defects and no advance in optic nerve cupping. This was achieved in 88.68% of the cases at 12 months follow-up. CONCLUSIONS: Deep sclerectomy is an efficient technique to control IOP, with few early post-operative complications. PMID- 12743844 TI - [Frequency doubling perimetry in terminal visual field defects]. AB - PURPOSE: To evaluate the Frequency Doubling Technology (FDT) in end-stage glaucomatous visual field defects. METHODS: FDT (C-20 threshold test) was performed in 22 consecutive patients presenting an end-stage visual field defect with 24-2 SITA (0 dB in more than 75% and less than 100% of the visual field). Comparisons of the percentage of non-abolished points, topographic correlation, test duration and global indexes were performed between C-20 and 24-2 SITA test. RESULTS: FDT sensitivity was 100%. C-20 test showed 13.7% more points with sensitivity greater than 0 dB compared to 24-2 test (P = 0.002). Two and a half less minutes were required for C-20 test performing (P<0.001). The mean value of Mean Defect was 11.48 dB better in FDT (P = 0,000). The mean value of the Pattern Standard Deviation was 5.37 dB (SD: 1.92) and 6.35 dB (SD: 3.61) for 24-2 and C 20 test respectively (P = 0.258). The inferotemporal quadrant was the best conserved in both perimetric strategies (Kappa agreement test value = 0.911; P<0.001). CONCLUSIONS: FDT showed greater ability than 24-2 test for end-stage visual field defects evaluation, with the advantage of being less time-consuming. Very good topographic correlation between both strategies was found. Further studies evaluating FDT reproducibility in severe glaucomatous visual field damage are necessary. PMID- 12743845 TI - [Ultrasonic biomicroscopic findings after spontaneous resolution of a traumatic cyclodialysis]. AB - CASE REPORT: We present one patient that received a blunt trauma in his right eye that resulted in ocular hypotension and hypotony maculopathy. UBM demonstrated the presence of a cyclodialysis. Several months later the patient experienced spontaneous improvement. A new UBM documented the presence of an anterior synechia closing the cleft. DISCUSSION: Ciclodyalisis is an infrequent problem. There are several treatment options, including conservative treatment, laser cyclopexy or surgical cyclopexy. Sometimes an spontaneous closure of the cleft may occur. PMID- 12743846 TI - [Proliferative radiation retinopathy]. AB - CASE REPORT: A thirty year-old-woman that had received radiotherapy three years before for a frontal glyoma consulted because of diminished visual acuity in her left eye. Examination showed a proliferative radiation retinopathy in the left eye and non-proliferative radiation retinopathy in the right eye that was confirmed by fluorescein angiography. The patient was treated with panretinal photocoagulation, and her visual acuity remained stable. DISCUSSION: Patients receiving cranial or neck radiotherapy should be followed for long periods of time because radiation retinopathy may appear many years after the treatment. Follow-up may permit early diagnosis of ischemic radiation retinopathy that can benefit from laser photocoagulation. PMID- 12743847 TI - [Intraocular cilia with macular pucker: clinicopathologic correlation]. AB - PURPOSE/METHODS: To report the pathological findings in a case of intravitreous cilia with macular pucker. We considered clinical factors to establish treatment. A pars plana vitrectomy and premacular membrane peeling were performed. Surgical samples were removed and studied with PAS, hematoxylin-eosin, Masson's Trichromic and immunohistochemistry. RESULTS/CONCLUSION: The histopathological examination disclosed the presence of cilia partially surrounded by a cuticle layer and internal limiting membrane with a celullarity increase. Surgical extraction is indicated in those cases in which the intraocular cilia are associated with any surgically treatable process or when it produces ocular inflammation or visual disturbance. PMID- 12743848 TI - [Severe retinal phlebitis in ocular bartonellosis]. AB - PURPOSE/METHODS: To report the clinical case of a 34-year-old male with atypical ophthalmic manifestations of cat-scratch disease (ocular bartonellosis), including an extensive retinal phlebitis, as well as the evolution of the clinical picture with treatment. RESULTS/CONCLUSIONS: The clinical diagnosis of ocular bartonellosis was serologically confirmed. Oral treatment with ciprofloxacine healed the phlebitis and the neuroretinitis, with a residual optic disk pallor. This case exemplifies the diversitiy of ocular manifestations of this disease. The authors recommend considering this condition in the differential diagnosis of posterior uveitis in young patients. PMID- 12743849 TI - [Bilateral carotid occlusion in young woman. Clinical and hemodynamic ocular results]. AB - CASE REPORT: We report the case of a 44-year-old woman, with a history of cigarette smoking (60 per day) and hyperthyroidism. She presented a bilateral ocular ischemic syndrome, and a bilateral occlusive carotid artery disease which showed reverse ophthalmic artery flow that resulted in a steal phenomenon, characterized by a shunt to the low resistance intracranial circuits and reduction of blood flow to the eyes. DISCUSSION: Young patients with severe bilateral occlusive carotid artery disease may be at greater risk of developing ocular ischemic syndrome, while this reverse flow decreases the patient's risk of stroke and death. PMID- 12743850 TI - [Cataract extraction seen by Rembrandt]. PMID- 12743851 TI - [Onchocerciasis]. PMID- 12743852 TI - [Multiple sclerosis: imaging, diagnostic criteria and differential diagnosis]. AB - Multiple sclerosis (MS) is the most common demyelinating inflammatory disease of the central nervous system (CNS), presenting with multifocal, disseminated inflammatory lesions referred to as plaques. Magnetic resonance imaging (MRI) typically depicts multiple, round to oval, circumscript lesions predominantly involving periventricular and subcortical white matter, brainstem and cerebellum. More recent investigations have demonstrated that the macroscopically visible plaques only present the tip of the iceberg: Already early in its course, MS causes neuroaxonal damage and diffusely involves the entire brain parenchyma including normal appearing white matter. These changes are reflected by strongly T1w hypointense lesions and atrophy of early onset, by reduction of the neuronal Marker N-acetylaspartate (NAA) on spectroscopy, by a decrease of the magnetization transfer ratio (MTR), by an increased in diffusibility and decreased anisotropy on diffusion-weighted imaging (DWI). MRI imaging is an important tool in the diagnosis of MS by revealing the characteristic spatial and temporal dissemination of the cerebral and spinal manifestations of this disease. Diagnostic criteria increase the diagnostic specificity and allow better differentiation from other diseases with multifocal white matter abnormalities. PMID- 12743853 TI - [Elastography: Quantitative imaging modality of the elastic tissue properties]. AB - The wide range of elastic tissue properties and the difference in elasticity of tumors and the adjacent tissues have provided motivation for developing elasticity imaging techniques. These research efforts have converged with the development of a new imaging modality, the so-called elastography, with the acquired and processed images referred to as elastograms. This modality was pioneered for ultrasonic imaging, which provides the majority of experience on this topic. MR-elastography has been developed in recent years. The published clinical data range from the characterization of prostate and breast cancer in oncology and dynamic examinations of skeletal muscle to intravascular elastic differentiation of vessel plaques. Advantages of the ultrasonic elastography are ubiquitous applicability and cost-effectiveness. MR elastography offers improved reconstruction and the possibility to assess potential anisotropic properties. The different approaches of elastography, their technical and clinical application and the limitations known so far are reviewed in this paper. PMID- 12743854 TI - Pulmonary balloon angioplasty of chronic thromboembolic pulmonary hypertension (CTEPH) in surgically inaccessible cases. AB - The clinical course of patients suffering from chronic thromboembolic pulmonary hypertension (CTEPH) depends on the distribution pattern of the thromboembolic material. In patients with thromboembolic findings in the central pulmonary segments pulmonary thrombendarterectomy (PTE) has excellent results and acceptable operative risk. This paper presents two surgically inaccessible cases that were successfully treated with balloon pulmonary angioplasty. Balloon angioplasty improved parenchymal perfusion, increased cardiac index (Delta CI +19.2 % [Case 1], and +15.4 % [2]), reduced pulmonary vascular resistance during follow-up (Delta PVRI -25.0 % [1] and -15.9 % [2]), and is discussed as an alternative treatment option for cases not suited for surgery. PMID- 12743855 TI - [The contribution of different postprocessing methods for multislice spiral CT in acute pulmonary embolism]. AB - PURPOSE: To investigate the value of different postprocessing algorithms for multislice spiral CT (MSCT) in diagnosing acute pulmonary embolism. MATERIALS AND METHODS: Forty-eight patients with suspected pulmonary embolism prospectively underwent MSCT using an 8-slice spiral CT. Using a confidence level on a three point scale, three radiologists reviewed 2-mm and 5-mm axial slices, 5-mm and 10 mm axial maximum intensity projections (MIP) and 2-mm coronal slices as well as interactively generated multiplanar reformatted images. A subsequent consensus reading of the primary 1.25-mm axial slices served as gold standard. ROC analysis was applied to the various vascular sections. RESULTS: The ROC analysis revealed a higher diagnostic accuracy of the 2-mm axial sections as compared to the 5-mm axial slices (Az = [0.988;0.976] vs. Az = [0.988;0.802]). Coronal and multiplanar reformations also showed excellent diagnostic accuracy (Az = [0.972;0.949] and Az = [0.997;0.951], respectively) and were significantly superior to the 5-mm axial slices through the segmental and subsegmental arteries (p=0.05). MIP showed the weakest diagnostic accuracy (Az = [0.967;0.802] for 5-mm MIP; Az = [0.879;0.781] for 10-mm MIP). CONCLUSION: Thin axial slices as well as coronal and multiplanar reformations are superior to thick axial slices in the diagnosis of acute pulmonary embolism. MIP is not suited for accurate diagnosis of pulmonary embolic disease. PMID- 12743856 TI - [ECG-triggered MDR-CT for the detection of pulmonary metastases]. AB - PURPOSE: Comparison of multidetector-row CT (MDR-CT) of the chest with and without ECG triggering for the detection of pulmonary metastases. MATERIALS AND METHODS: Fifty patients with malignant tumors underwent CT of the chest. The unenhanced phase was performed with ECG-triggered MDR-CT and the contrast enhanced phase with helical MDR-CT. The ECG-triggered and standard helical scans were interpreted in separate sessions, with the analysis determining the number and demarcation of the intrapulmonary nodules and the delineation of the mediastinal structure (rated 1 = excellent to 5 = poor). RESULTS: ECG-MDR-CT images detected 38 % more pulmonary nodules than MDR-CT. The detection rate for tumors < 5 mm was 64 % higher in ECG-triggered scans. The median demarcation rating of all pulmonary findings was 2 for ECG-MDR-CT and 3 for MDR-CT. The median demarcation rating of tumors < 5 mm was 3 for ECG-MDR-CT and 4 for MDR-CT. Mediastinal structures were better delineated by ECG-triggering. The median demarcation rating of the vessels and left bronchus was 2 for ECG-MDR-CT and 4 for MDR-CT. Detection rate and demarcation rating of pulmonary tumors and the delineation of mediastinal structures were not significant different for either CT technique. CONCLUSION: Our study indicates superiority for ECG-MDR-CT. PMID- 12743858 TI - [Teleradiology for emergency cranial computed tomography]. AB - PURPOSE: We report our experience with the teleradiologic service provided by a center hospital (CH) for emergency cranial computed tomography (CCT) in two regional hospitals (RH) during a 12-month period. The clinical and economic impact of teleradiology will be discussed as well as the acceptance by the clinicians of the regional hospitals. MATERIAL AND METHODS: In 2001, 213 CT-scans in 202 patients were performed and reported using teleradiology. Teleradiologic and final medical diagnosis were analysed by the medical reports. The transfer of the patients to a CH and their further treatment were checked. The referring physicians in the RH were asked to complete a questionnaire about the teleradiological support. RESULTS: 18 (9 %) patients had to be urgently transferred to a CH based on the CT findings in the teleradiological reports. 24 patients (11 %) were transferred to a center hospital during further treatment. 80 % of patients were treated in the RH. CONCLUSION: CT data transfer is reliable and efficient. Teleradiology reduces costs by avoiding unnecessary transport of patients for diagnostic procedures. Teleradiology improves patient management by reducing time from admittance to decide about further treatment. Teleradiology enables rapid selection of the best-equipped center hospital for the management of the patient's disease. Teleradiology is well accepted by the physicians in the RH. In the RH, teleradiology has become an indispensable standard procedure in the emergency diagnostic workup of cerebral trauma patients and in patients with acute unclear neurological symptoms. PMID- 12743859 TI - [Clinical relevance of cranial CT under emergency conditions--basic neuroradiologic investigations]. AB - PURPOSE: To determine the diagnostic accuracy of cranial CT under emergency conditions. MATERIAL AND METHODS: Four hundred cranial CT examinations (cerebrum, petrosal bone, fascial bone, sinuses, orbita) were evaluated retrospectively and correlated with the initial neurologic examination, the discharge and surgical diagnoses, and the results of other imaging modalities (conventional X-Ray, MRI, angiography). The average age was 49 (2 to 89) years, with 42 % male and 58 % female patients. The discharge diagnosis was defined as the gold standard, assuming that clinical examinations and laboratory and imaging studies were maximized to establish the diagnosis, together with any possible surgery. RESULTS: Overall sensitivity, specificity and diagnostic accuracy was 85 %, 70 % and 82 % for cranial CT, 66 %, 45 % and 61 % for the initial neurologic examination, 79 % and 29 % for acute cerebral infarction, and 65% and 13% for intracranial bleeding. Correlation of the CT diagnoses to the discharge and surgical diagnoses was high (82 % and 80 %, respectively; p < 0.01), to the diagnosis of conventional x-ray moderate and to the diagnoses of angiography and MRI unsatisfactory (74 % and 68 % and 60 %, respectively; p > 0.05). Craniotomy was performed in 47 of 400 (12%) patients after cranial CT, within 12 hours in 29, in 12 to 24 hours in 8, and after 24 hours in 10 patients. The risk for intracranial bleeding in patients with a skull fracture was increased by a factor of 3.3. CONCLUSION: The high diagnostic accuracy of cranial CT in establishing a precise, timely and reproducible diagnosis confirms the important role of CT as primary diagnostic method under emergency conditions. PMID- 12743861 TI - [MRI of the lateral ankle ligaments: value of three-dimensional orientation]. AB - PURPOSE: To determine the three-dimensional orientation of the lateral ankle ligaments with MRI. MATERIALS AND METHODS: Twenty healthy volunteers without previous injury to the ankle were included in the study. With the right ankle in the normal anatomic position stabilized in a splint, coronal T2-weighted spin echo sequences (TSE) were obtained. The three-dimensional orientation was determined by placing paths through the ligaments and by measuring the angles between corresponding tangents and the three main imaging planes. RESULTS: Using the calculated angles, full-length visualization of the lateral ligaments of the ankle was achieved. The angles deviating from the axial imaging plane were 18.0 degrees for the anterior talofibular ligament, 52.3 degrees for the calcaneofibular ligament and 28.2 degrees for the posterior talofibular ligament. CONCLUSION: MRI enables the exact determination of the three-dimensional orientation of the lateral ankle ligaments. Orienting the imaging planes according to the calculated angular deviation allows the full-length visualization of the ligaments and is the basis for optimal imaging of the lateral ankle ligaments. PMID- 12743860 TI - [Functional CT imaging: Load-dependent visualization of the subchondral mineralization by means of CT osteoabsorptiometry (CT-OAM)]. AB - PURPOSE: Functional computed tomography for visualization and quantification of subchondral bone mineralization using CT osteoabsorptiometry (CT-OAM). MATERIALS AND METHODS: Tarsometatarsal (TMT) and metatarsophalangeal (MTP) joints of 46 human hallux valgus (HV) specimens were examined (sagittal 1/1/1 mm) on a single slice CT scanner SCT (Somatom Plus 4, Siemens AG). Subchondral bone pixels were segmented and assigned to 10 density value groups (Delta 100 HU, range 200 - 1200 HU) the pixels using volume rendering technique (VRT). The data analysis considered the severity of HV as determined by the radiographically measured HV angle (a. p. projection). RESULTS: CT-OAM could generate reproducible densitograms of the distribution pattern of the subchondral bone density for all four joint surfaces (TMT and MTP joints). The bone density localization enables the assignment to different groups, showing a characteristic HV-angle-dependent distribution of the maximum bone mineralization of the load-dependent densitogram (p < 0.001). CONCLUSION: CT-OAM is a functional CT technique for visualizing and quantifying the distribution of the subchondral bone density, enabling a noninvasive load-dependent assessment of the joint surfaces. Load-dependent densitograms of hallux valgus specimens show a characteristic correlation with an increase of the HV-angle. PMID- 12743862 TI - [Evaluation of a hemostatic device with percutaneous collagen application (VasoSeal) compared to a mechanical compression system (Compressar) after transfemoral catheterization of patients suffering from arterial occlusive disease]. AB - PURPOSE: Comparison of the efficacy of VasoSeal and a mechanical compression system (Compressar) for percutaneous hemostasis after femoral arterial catheterization of patients with arterial occlusive disease. MATERIALS AND METHODS: 60 patients underwent either diagnostic angiography or interventional procedures. The level of anticoagulation, blood pressure, and activation clotting time were recorded, and the time to hemostasis after sheath removal was measured. VasoSeal application was considered "successful" if the compression time was less than two minutes. On the subsequent day as well as 4 months later, color coded Doppler ultrasound was performed to register treatment success and potential (late) complications. RESULTS: 57 patients qualified for inclusion in this study. In 21 of the 26 patients who underwent the procedure with the VasoSeal, immediate hemostasis was achieved within 1.75 minutes. In all 31 patients who had the Compressar applied, hemostasis was successful with a mean compression time of 17.4 minutes. Thus, VasoSeal significantly reduced hemostasis time irrespective of anticoagulation status, but it had a much higher incidence of minor local complications (bleeding, hematoma) compared to the control group (34.6 % vs. 5.8 %). The technical success was lower with VasoSeal than with Compressar (81 % vs. 100 %). Both groups had no severe or late complications. CONCLUSION: According to our results, VasoSeal does not provide a suitable alternative compared to the effective, safe and cheap application of Compressar as a hemostatic device. PMID- 12743863 TI - [Interstitial photodynamic laser therapy for liver metastases: first results of a clinical phase I-study]. AB - PURPOSE: Development and evaluation of a new photodynamic treatment technique for the laser therapy of liver malignancies MATERIAL AND METHODS: The combination with new catheter systems enables the use of the photodynamic therapy (PDT) to treat also tumors in parenchymal organs. So far it is mainly used to treat superficial or endoluminal tumors. The presented study is part of a multicenter phase I-study. We treated 5 patients with colorectal liver metastases with the new photosensitizer SQN 400 and following interstitial photodynamic laser treatment. Evaluation of tumors were performed by contrast-enhanced CT scans. RESULTS: In the contrast enhanced CT scans the development of a complete necrosis within a radius of 1 cm around every single fibre could be shown. Additional the ablation of tumors with the combined use of several fibres is possible. Severe complications or toxicities were not observed. CONCLUSION: The photodynamic laser therapy of liver malignancies is a minimal invasive procedure with little side effects which produces sharply defined yet small volumes of necrosis. PMID- 12743866 TI - [Minimally invasive therapy of adrenal gland metastasis]. PMID- 12743864 TI - [MR-guided biopsies of undetermined liver lesions: technique and results]. AB - PURPOSE: To evaluate safety and precision of liver tumor biopsies performed in an open low field system using different sequence techniques. MATERIALS AND METHODS: In 47 patients with liver tumors, MR-guided biopsies were performed in a low field system (0.2 Tesla, Magnetom Open, Siemens) using two different sequences. The procedure was monitored with T1-weighted FLASH sequences (TR/TE = 100/9; 70(3)) in all patients and with FISP-Rotated-Keyhole-sequence (TR/TE = 18/8; 90(3)) in additional 20 patients. After positioning of the needle tip in the tumors, 166 biopsy specimens were acquired with 16 G cutting needles (Somatex*). The diameter of the biopsied lesions ranged from 1 to 10 cm (mean diameter 3.2 cm). Visibility of the needles and precision of the biopsies were evaluated. RESULTS: All interventional biopsies were performed without vascular or organ injuries. Adequate specimens for histologic interpretation were obtained in 42 cases (89.3%). The biopsy results were non-specific in 2 patients (4.2%) and the lesions missed in 3 patients (6.3%). Mean in-room time was 35 minutes and the intervention time was 8.3 minutes. T1-weighted FLASH images proved optimal for confirming needle-tip placement during the biopsies or punctures. Organs, tumors and vessels were easily identified. The FISP sequence proved to be inferior in visualizing vessels and tumors. CONCLUSION: MR-guided liver biopsies are safely and precisely performed using T1-weighted FLASH-sequences with sufficient visualization of the lesions and might be complementary to US- or CT-guided biopsies. PMID- 12743865 TI - [Transarterial chemoperfusion of inoperable pancreas carcinoma and local recurrence]. AB - BACKGROUND: To evaluate the side effects and tumor response of a locoregional transarterial chemoperfusion with Mitomycin C and Gemcitabine in advanced pancreatic cancer. MATERIALS AND METHODS: Between October 2001 and August 2002, 16 patients were treated in 80 transarterial chemoperfusion sessions with a mean of 5 sessions per patient and a pause of 4 weeks between the sessions. The chemotherapy consisted of a combination of Mitomycin C (8.5 mg/m 2) and Gemcitabine (500 mg/m 2), administered within 1 hour. The tumor response was evaluated by MRT and CT and the quality of life by a patient questionnaire. RESULTS: All patients tolerated the procedure well. The therapeutic outcomes were as follows: in 50 % (n=8) of patients, minor response with a decrease in size of the primary tumor and metastases up to 20 %; in 25 % (n = 4) of patients, stable disease; and in 25 % (n = 4) of patients, progressing metastases with stable primary tumor. Eleven patients (68.75 %) reported a good quality of life without impaired performance of their daily activities. Side effects were moderate and reversible between the treatment cycles. After one year 25 % of the patients were still alive. CONCLUSION: Transarterial chemoperfusion is a minimally invasive treatment for pancreatic carcinomas and local recurrences with little side effects and can be delivered on an outpatient basis. It might be indicated as potential neoadjuvant treatment before surgery or as palliative treatment to provide clinical benefits and to improve the quality of life. PMID- 12743868 TI - [Foreign body ingestion in childhood after dental care with rare sequela of perforation of the small intestine and pancreatic injury]. PMID- 12743867 TI - [Secondary liver involvement in multiple myeloma: diagnostic imaging with magnetic resonance tomography (MRI)]. PMID- 12743869 TI - [Is gastric and duodenal ulcer drug-treatable or surgical disease?]. PMID- 12743870 TI - [Bilateral carcinoma of the breast]. AB - From 1990 to 1996, a total of 223 patients, with breast cancer, were treated. Out of these patients, 7 (3.04%) developed a second primary carcinoma in the controlateral breast. Two (28.5%) had a synchronous manifestation while the remaining 5 (71.5%) had a metachronous manifestation. The seven patients were followed up for periods between 4-8 years: 80% of the metachronous tumours occur within 5 years of follow-up. The 7-year actuarial survival was 100% in the two synchronous tumours and 60% in the five metachronous tumours (two patients died on account of intervening diseases). The prognosis does not depend on the length of the interval between the development of the two carcinomas but it is in relation with the anatomo-clinical state, to the adequate treatment of each primary tumour and to the rigorous clinical-instrumental follow-up, in which the mammography plays a fundamental role. PMID- 12743871 TI - [Primary cancer of the thyroid: natural history and therapy principles]. AB - Thyroid primary cancer involves 1.5% of malignant tumors and it causes yearly 0.5% cancer mortality. It is usually related to eu- or hypothyroidism, whereas it is rarely associated with hyperthyroidism. The Authors deal with controversial issues concerning thyroid oncogenesis and surgical strategies. However, they have not ignored epidemiological data and clinical features most peculiar to various neoplastic histotypes. They also report on their own experience with 156 cases of thyroid CA, out of 2.510 surgically treated thyroid diseases, which were being investigated from 1969 to 1998. The Authors recognize that an appropriate approach to thyroid neoplastic diseases arises from a careful evaluation of all parameters (T, N, M, histotype, patient's, age, intraoperative assessment etc.) characterizing their polymorphism. PMID- 12743872 TI - [Thyroid carcinoma in childhood]. AB - Thyroid carcinoma in childhood is very rare. The Authors report the features of this neoplasm versus other period of the life. There were no undifferentiated type of neoplasm in 69 cases. Almost all the tumors were well differentiated epithelial types especially papillary and only few cases were non epithelial differentiated tumors. Prognosis, that is better on childhood than other ages, depends, besides surgical indications (total thyroidectomy with lymphadenectomy for epithelial types of neoplasm to metabolic radiotherapy even if metastatized. Medullary carcinoma that as after associated to other endocrinopaties has the worst prognosis; total thyroidectomy with bilateral lymphadenectomy as not enough to avoid recurrence that does not responds to complementary treatment. Thyroid surgery in the child exposes to a major risk of iatrogenic lesions than the adult because of anatomical characteristics. PMID- 12743873 TI - [Surgery of the thyroid in short-stay hospitalization]. AB - New trends and improvements in health care system allow to choose short hospital stay even on major surgical operations. Authors report their experience on 200 moderate or good operative risk patients who underwent operation for thyroid disease from 1993 to 1998. Short hospital stay program was achieved after 48-72 hours under observation in 193 patients (96%) who underwent subtotal, total and total with lymphadenectomy, thyroidectomy. All the patients, informed on detail were kept under close phone contact after discharge to be able to take part if need be. PMID- 12743874 TI - [Differentiated cancer of the thyroid: prognostic aspects]. AB - Ten-year span survival from differentiated thyroid cancer has been reported as ranging around 90%. Prognostic factors to be regarded as positive elements are young age, lack of lymph node and hematogenous metastasis, size < 4 cm, and low grading. The Authors investigate those features that have seemed to affect morbidity rate and long-term survival from differentiated thyroid cancer (T, N, M, age, sex, ploidy, benign thyroid lesions, etc.). The Authors deem the appreciation of such prognostic factors as being conducive to a better selection of treatments for any patient in terms of survival span and quality of life. PMID- 12743875 TI - [Preoperative staging of colorectal neoplasms: importance of the echo-endoscopy]. AB - The A. A., after having confirmed how the timely diagnosis of cancer of the Sigma Rectum section is the result a complicated and articulate diagnostic iter, describe the advantages of using the ultrasonography-endoscopy. Therefore they relate their experience, comparing the outcome of this research with that of the TC images and above all with the histo-pathological surveys on the removed sechions. They conclude affirming that a higher percentage of the diagnostic care is a clear evolution in the correct pre-operating stage of these neoplasias. PMID- 12743876 TI - [Gastrointestinal metastasis of melanoma: a case study]. AB - The Authors report on a case of multiple metastases located in the jejunum (where it caused intussusception), the omentum, and retroperitoneum having originated from a malignant melanoma with subungual primary site on first finger of left hand. After thoroughly surveying the specific literature, the Authors deal in more detail with current treatments and therapeutic prospects for this kind of neoplastic disease. PMID- 12743877 TI - [Subclavicular localization of Castleman's disease]. AB - Castelman disease is a rare disease of lymph nodes. There are 2 pathological types: hyalo-vascular asymptomatic form and plasmacytic symptomatic form. Authors report a case of Castelman disease of a subclavicular lymph nodes that underwent surgical operation. Ethiopathogenesis and treatment of the disease are reported. PMID- 12743878 TI - [Day-surgery treatment of pilonidal sinus]. AB - Authors discuss ethiopathogenus of pilonidal disease after a historical background and support acquired-congenital theory. Open and closed treatment are reported on the acute and chronic phases, highlighting economical advantages of closed method. Carbon-dioxide laser on Authors experience was very useful to prevent complications and to shorten healing time in open method. PMID- 12743879 TI - Dietary intake of ten key nutrients for public health, United States: 1999-2000. AB - This report presents dietary intake estimates of 10 nutrients for the U.S. population by sex and age groups. Nutrient intakes are estimated from one 24-hour dietary recall interview conducted in the National Health and Nutrition Examination Survey, 1999-2000. Population means, medians, and standard errors of the mean are weighted to produce national estimates. Assessment of dietary intakes is an important part of monitoring the nutritional status of the U.S. population. PMID- 12743880 TI - Public Private Partnership (PPP) for essential health services. An innovative approach for optimum and efficient use of public and private resources. PMID- 12743881 TI - PPP insights in South Africa. AB - After functioning for some time in an increasingly regulated and structured environment in dealing with the private sector in South Africa, it was important to Government, to carefully review the terminology used in this evolving playing field. As the definitions and mechanisms impacting on this form of interaction became clear, it was essential to find a broader definition to encompass all forms of commercial intervention between the two sectors. In preparation for the first South African National Health Summit during 2001, the term public private interaction became a general term used in this context. In the South African healthcare sectors this term is used specifically to indicate that all forms of interaction between the two sectors should be considered, rather than merely focussing on specific Public Private Partnerships (PPPs), that have a much more narrow definition. Recent health policy documents in South Africa all stress four key goals--equity, coherence, quality of care and efficiency--which provide a useful basis for decision-making about PPIs. The range of public-private interactions that may support or constrain the South African health system's development are set within the overall public/private mix of the country. In developing an equitable, efficient, coherent and high quality health system in South Africa, there is considerable potential for constructive engagement (collaboration and co-operation) between the public and the private health care sectors. Both sectors should embrace this opportunity and therefore it is useful to propose some basic guidelines for engagement based on the vision and goals of the national health system. In deciding whether or not to pursue any new PPI within the health sector, or in evaluating whether an existing PPI should continue or be revised, it is necessary to assess its merits in relation to the achievement of health system goals. PMID- 12743882 TI - Private health care sector investment in Brazil: opportunities and obstacles. AB - The Brazilian health system is based upon the constitutional right formulated in 1988, according to which health is the peoples' right and duty of the State. So being, it is essentially the government's responsibility, expressed in the so called Sistema Unico de Saude--SUS (single health system) Since its creation, however, it admits the existence of a supplementary health system, left to the private sector. In general terms, the public system is considered unsatisfactory in the services it renders. Its resources are distributed heterogeneously, favoring centers of advanced medical practice, to the detriment of basic health care. The supplementary system is considered of better quality, however with great variations and frequent accusations of being essentially profit driven, instead of being driven to the needs of the assisted population. The growing search for health plans is a direct consequence of the image perceived by the population regarding the quality and accessibility of the public services, as well as of the peoples' growing consciousness of their needs, rights and duties as citizens. The need for continuous quality improvement and cost reduction offers numberless opportunities for actions and investments. Initiatives to identify and implement the best medical practices, medical guidelines and actions are essential regarding those illnesses which are most frequent, of higher cost and of greater risk. Health plans and healthcare providers will necessarily have to focus on their common client. Therefore, organizations must be created in order to develop initiatives aimed to the quality of patient care, as well as to the collection and dissemination of data regarding the production and results of the main service providers. Consequently, immense opportunities are being opened for investments in the area of Information Technology, collection, analysis, and data dissemination. This paper analyses the main trends in the Brazilian health sector and from the data and tendencies, draws various conclusions on the opportunities and barriers for private investment. PMID- 12743883 TI - [Hospital emergency and health system organization in France]. AB - There is no legal definition of the term "emergency" in France. Given the vagueness of this term, a law was passed on 6 January 1986 in France defining the principle of urgent medical assistance as "providing the sick, the wounded and pregnant women, wherever they are, with the appropriate emergency care to meet their condition". The Ethical Medical Code of 6 September 1995 (Art. 77) specifies that "regarding constant availability of health care, the duty of every doctor is to take part in the day and night on-call rota". To understand the emergency care system in France, it should be stressed that the health system is a joint system, with a private sector and a public sector jointly organizing and providing health care. The patient can access either system as he chooses. PMID- 12743884 TI - Health insurance and the poor in low income countries. AB - This short paper is not by an economist. It is by a public health doctor who has spent the last ten years working with development agencies and governments to strengthen public health systems in poor and middle-income countries. It is becoming increasingly obvious that while this still needs to be done, the focus of attention must shift to private expenditure and private providers. In most poor and middle income counties--even the most aid dependant--by far the biggest source of finance in the health sector is out of pocket expenditure and the poor in many of these countries spend disproportionately more than the rich as a % of household income on health care. This expenditure is primarily in the private sector or on unofficial user fees in the public sector. This expenditure does not protect the poor from the economic costs of catastrophic illness nor do they appear to get good value for treatment in the primary care setting. This paper is not a detailed synthesis of current knowledge but an overview for those who are not specialists in this area. It gives references to key publications, which have done that synthesis, and draws from them. PMID- 12743885 TI - Tradition and modernity in upgrading the new "Meyer" Children Hospital in Florence. AB - The article intends to illustrate the methodology and the design experience by which the architectural team (C.S.P.E.--Florence + Anshen and Allen--San Francisco) has succeeded in satisfying the competition requirements proposing a balanced combination of a totally new block strictly integrated with the upgraded existing building. Specific emphasis has been attributed to the environmental performances of interior and exterior spaces in order to create a facility characterized by "children dimension". The presence of a monumental garden area, all around the building, has been used as a generator-input of the functional organization and, in this sense, has resulted particularly successful for the Public Client Organization. PMID- 12743887 TI - Weaving a care network. PMID- 12743886 TI - Leadership development: prerequisite for successful public-private partnership. New challenge for medical schools: graduates' outcome competences in health leadership. AB - Medical education stands at the doorstep of profound change, forced to step into an uncertain and potentially hostile new environment. These changes have nothing to do with scholar self-reflection, but rather are a direct consequence of the process of globalization visible also in medical education and the revolution in health care financing for which we use the general term "managed care". On one hand, globalization has penetrated different areas of our life, among others including science, public health and medicine which is a global profession. Medical knowledge, research and education have traditionally crossed national boundaries. Many aptitudes of physicians are universal, as well as the core competencies required by physicians throughout the world. The question is "what kind of core or essential competences are required for "global physicians". Besides medical knowledge and basic clinical skills, professionalism, interpersonal communication skills, context of care, information management, understanding a health system, etc. also need to be addressed. On the other hand, there is a growing concern that the corporate transformation of medical care, especially related to the impact of the changing and more business-oriented health care system, may lead to the decline and death of traditional professional values, such as fidelity, altruism, confidentiality, and integrity. No doubt, there is an inherent clash of values between business and medicine. However, as business interests have already gained an important place in medicine, we can hope that physicians progressively adopt a business mentality without losing professional virtue. While occurring changes inevitably bring some sense of loss, it also brings an opportunity to help reshape medical education to better meet the health needs of society. PMID- 12743888 TI - Broad canvas needs fine detail. PMID- 12743889 TI - Reaching behind the bars. PMID- 12743891 TI - How older people manage stress. PMID- 12743890 TI - Family carers' experiences: reflections on partnership. PMID- 12743892 TI - Tuberculosis and older people. AB - Tuberculosis is a disease with a long history and although concerted efforts have been made to eradicate it, cases have increased in numbers since 1985. This article considers symptoms, diagnosis and treatment and explains how older people with TB present particular difficulties for those caring for them. PMID- 12743893 TI - Assessing blood results in older people: haematology and liver function tests. PMID- 12743894 TI - Interim 2001-based national population projections for the United Kingdom and constituent countries. AB - This article describes new 2001-based national population projections which were carried out following the publication in September 2002 of the first results of the 2001 Census. These "interim" projections, carried out by the Government Actuary in consultation with the Registrars General, take preliminary account of the results of the Census which showed that the base population used in previous projections was overestimated. The interim projections also incorporate a reduced assumption of net international migration to the United Kingdom, informed by the first results of the 2001 Census and taking account of more recent migration information. The population of the United Kingdom is now projected to increase from an estimated 58.8 million in 2001 to reach 63.2 million by 2026. The projected population at 2026 is about 1.8 million (2.8 per cent) lower than in the previous (2000-based) projections. PMID- 12743896 TI - How important are intergenerational cycles of teenage motherhood in England and Wales? A comparison with France. AB - Teenage fertility has fallen substantially in every Western European country except the United Kingdom. This article examines the hypothesis that repetition of teenage motherhood from mother to daughter is a major cause of the UK being the exception. A simple demographic model of fertility across generations is estimated with comparable data from England and Wales and France. The main finding is that mother-daughter repetition can account for only a minor part of the total difference in teenage childbearing between the two countries, especially over the long term. The higher teenage childbearing in England and Wales of those whose mothers began childbearing after their teenage years dominates. PMID- 12743895 TI - Higher qualifications, first-birth timing, and further childbearing in England and Wales. AB - This article examines how strong the association is between the obtaining of higher educational qualifications and later entry to motherhood, and how these are associated with levels and pace of second and subsequent childbearing. Data from the ONS Longitudinal Study are used to estimate these associations for women born in England and Wales between 1954 and 1958. Average age of entry to motherhood is found to be five years later for women with higher qualifications than for those without. Increasing age of motherhood is always associated with a lower likelihood of going on to have another child, but the decline with age is less pronounced for women with a higher qualification. Moreover, for any given age of childbearing, mothers with a higher qualification are more likely than those without to have another child, and are more likely to do so quickly. PMID- 12743897 TI - Oncology update. PMID- 12743898 TI - The Georgia Cancer Center for Excellence. AB - The Georgia Cancer Center is designed to encourage academic study of cancer health disparities and try to develop efficient ways to provide state-of-the-art care. The center has leveraged resources from the public and private sector to create a center for research and clinical excellence. The Georgia Cancer Center will eventually offer unique phase I and phase II drug trials and unique research treatments. Patients from throughout the state will be eligible for such care with referral from their local physicians. PMID- 12743899 TI - Imatinib: the promise of a "magic bullet" for cancer fulfilled. PMID- 12743900 TI - New surgical techniques for the primary management of breast carcinoma. PMID- 12743901 TI - New options in the prevention and treatment of breast carcinoma. PMID- 12743902 TI - New developments in the management of ovarian cancer. PMID- 12743903 TI - Chemoprevention in lung cancer. PMID- 12743904 TI - Genomic medicines for oncology: early lessons. PMID- 12743906 TI - The role of self-administered plaque control in the management of periodontal diseases: 2. Motivation, techniques and assessment. AB - The delivery of oral hygiene advice is a crucial component of the management of patients susceptible to periodontal disease. However, the complexity of the issues surrounding such advice is frequently underestimated. It is not simply a question of manual dexterity; many factors influence compliance and motivation, including lifestyle, beliefs, attitude and understanding, and such factors need to be taken into account in order to effect permanent change in a patient's habits. This article reviews these issues and examines oral hygiene techniques and methods of assessment. PMID- 12743905 TI - Monoclonal antibody therapy of non-Hodgkin's lymphoma: the Rituximab story. AB - The advent of monoclonal antibodies targeted at tumor associated or tumor specific antigens provides a novel approach for the treatment of a broad range of malignancies. Such targeting agents can either be used as unlabeled molecules to induced antibody dependent cytotoxicity, or complement mediated lysis and/or apoptosis, or be conjugated with cytotoxic moieties such as toxins and/or radioactive nucleids. The applicability of this strategy has been expanded beyond the treatment of NHL as demonstrated by the clinical application of the chimeric monoclonal antibody directed at the Her2/neu receptor in breast cancer, the epidermal growth factor receptor in colon and head/neck cancer, and the vascular endothelial growth factor in lung and colon cancer, among others. In a variety of settings this strategy demonstrates clinical anti-tumor activity in patients who have failed to respond to or are refractory to conventional cytotoxic chemotherapy. Furthermore, given the modest toxicity of these naked antibodies, they avail themselves as ideal partners for combining with conventional chemotherapy to produce results that often appear to be greater than additive. With the discovery of newer targets and means to manipulate the immunoglobulin molecule to generate tailor made antibody fragments, the field of antibody based targeted therapy of cancer appears to be on the threshold of making major strides in the fight against cancer. PMID- 12743907 TI - Low-shrink monomers for dental restorations. AB - The main disadvantages of resin-based composites (RBCs) for use in load-bearing posterior restorations include the polymerization shrinkage following curing and inadequate wear resistance in service. These properties are largely influenced by the monomer system and research is currently being undertaken to decrease polymerization shrinkage and improve resin wear characteristics in an attempt to increase RBC restoration longevity. The scope of the current review will identify the development of resin-based restoratives, indicating the reported advantages and disadvantages of resin types routinely used in dental practice today and review the most recent advancements in resin technology. PMID- 12743908 TI - Does the GDP need to know about IOTN? AB - 'Two-tier dental systems planned'; 'Children in need of orthodontic treatment to go without'... recent scaremonger headlines from consumer reports exaggerate the latest potential effects of this government's plans to target financial resources at those in greatest need. This article aims to provide a background to occlusal indices and provide general dental practitioners (GDPs) with an insight into the use of the Index of Orthodontic Treatment Need (IOTN), which it is understood may be introduced into the General Dental Service in the future. PMID- 12743909 TI - Correction of the ptotic chin. AB - Where there is extensive stripping of the mentalis muscle from the mandible during vestibuloplasty procedures the chin tissues often become lax and droop, giving an unsightly ptotic defect. If this soft-tissue defect is combined with excessive chin tissue an excisional surgical procedure is required. This article describes a surgical procedure to produce an acceptable chin profile. PMID- 12743910 TI - The oral health of a group of prison inmates. AB - The oral health of prison inmates in England has come under increased scrutiny with the arrival of joint responsibility between the Home Office and the NHS for prison healthcare. This brief study indicates very high levels of oral disease amongst a group of prisoners attending for treatment in an English prison. Further study of the oral health of prisoners seems timely, as does the exploration of effective oral health promotion for this group of people. PMID- 12743911 TI - The distal extension base denture. AB - The distal extension base denture may be indicated in situations in which the edentulous area to be restored is without a terminal abutment tooth. There may be significant challenges in providing a prosthesis with sufficient support and retention to make it comfortable without damaging the intra-oral tissues. This can be a greater problem in the mandible as the denture-bearing area is usually much smaller than in the maxilla. This paper considers how distal extension removable prostheses can be designed to restore edentulous spaces. PMID- 12743912 TI - Selection for vocational/general professional training--the Dundee undergraduate experience. AB - Vocational training has been mandatory within the dental profession since 1993 but, despite this, there is continued disquiet surrounding the application process. This paper collates and summarizes the experiences of applicants from one dental school, to inform debate and assist in the continued evolution of the process of application for vocational training. PMID- 12743913 TI - Making occlusion work: I. Terminology, occlusal assessment and recording. AB - This is the first paper in a two-part series reviewing some of the relevant theoretical aspects of occlusion and describing its application in clinical practice. This article discusses terminology, clinical examination of the occlusion, articulators and interocclusal records. PMID- 12743914 TI - Choosing a career in dentistry. PMID- 12743915 TI - Choosing a career in dentistry. PMID- 12743916 TI - The last post. PMID- 12743917 TI - Case study--foreign object in palate. PMID- 12743918 TI - Preserving the vital pulp in operative dentistry. PMID- 12743919 TI - Physical signs for the general dental practitioner. Case 3. Spider naevi and jaundiced sclera. PMID- 12743920 TI - Penetrating brain injury in military conflict: does it merit more research? PMID- 12743921 TI - Common medical disorders related to diving--prevention, diagnosis and fundamentals of treatment. Part 1: Diving disorders that do not require recompression. PMID- 12743922 TI - The pipe bomb: a modern terrorist weapon. AB - The use of pipe bombs by terrorist organisations around the world has risen dramatically. The pipe bomb is simple and easy to make and details of bomb construction are freely available from the Internet. Components can be found in most hardware stores and thus give little forensic information. Despite the increasing use of pipe bombs, clinicians may be unfamiliar with the type of injuries they cause. This paper describes two incidents of pipe bomb explosions and illustrates the type of injuries sustained by both terrorists and victims. PMID- 12743923 TI - The case of Edward Revere Osler. AB - In today's military medical environment interoperability is a regular occurrence. During the First World War a small group of eminent medical men from England and America combined their skills to endeavour to save the life of the son of another eminent physician. PMID- 12743924 TI - A drill-free bone screw for intermaxillary fixation in military casualties. AB - Drill-free bone screws are a simple and quick method of establishing intermaxillary fixation requiring a minimum amount of specialist training or equipment. These screws offer significant advantages over other methods of intermaxillary fixation and are well suited for use in military casualties. PMID- 12743925 TI - Command in a field hospital. AB - This paper examines the challenges involved in commanding a field hospital. There are frequent, dynamic tensions between the military culture that is based on a task-focussed, hierarchical structure and the clinical culture that is based on flat, process-focussed, multidisciplinary teams. The paper outlines the cultural environment of the field hospital and then examines the deployment sequence whereby a functioning clinical facility may be created from a group of disparate individuals. There are a number of tools that may assist with this including the personality of the Commanding Officer, individual skills, the creation of an organizational identity and the choice of command structure. PMID- 12743926 TI - Sexuality and soldiery combat & condoms, continence & cornflakes. AB - The results of a short sexual health study of 75 male soldiers undertaking humanitarian aid relief in Africa are presented in an historical context relating to sexuality and soldiery. PMID- 12743927 TI - Combined UK/US field hospital management of a major incident arising from a Chinook helicopter crash in Afghanistan, 28 Jan 2002. AB - On Monday, 28 January 2002, a US Army Chinook helicopter crashed on landing in Afghanistan. Sixteen casualties were airlifted from the scene for treatment at the US Army 274th Forward Surgical Team and the British 34 Field Hospital Troop at Bagram airfield before aeromedical evacuation out of Afghanistan. This was the largest mass casualty incident to be dealt with in a combined fashion by the British and American medical services in Afghanistan during the initial months of Operation ENDURING FREEDOM. It illustrated how multinational surgical teams can successfully manage such incidents by following common and agreed protocols. The lessons learned are relevant to any combined operations in the near future. PMID- 12743928 TI - Use of atypical anti-psychotics in the management of post-traumatic confusional states in traumatic brain injury. AB - The use of atypical anti-psychotics (AAP) in the treatment of organic neuropsychiatric syndromes is little reported. We present a case of post traumatic delirium with delusions treated with Risperidone and discuss the use of AAP's in this situation. PMID- 12743929 TI - A case of a uniocular pterygium related to an unusual occupation. PMID- 12743931 TI - Self-assessment exercises in pre-hospital emergency trauma care. PMID- 12743930 TI - Battlefield advanced trauma life support. PMID- 12743932 TI - Military surgery or surgery in the military? PMID- 12743933 TI - Preparing for the FFAEM examination. AB - While the pass rate for this examination is relatively high compared to previous postgraduate examinations such as membership or fellowship (of whichever college) it should still not be underestimated. It is true that those who have been through a recognised training scheme should pass and failure reflects upon the scheme as well as the candidate, but the consequences of failure are high and fall directly only upon the candidate. Thorough preparation will immensely improve the chances of passing first time and finishing the critical topic reviews well in advance is thoroughly recommended (it would be ideal to do one each year during the first four years of training and then choose the best two for the examination). This will leave time to go through the management books and as many short clinical questions books as possible, those designed for MRCP are especially helpful. While the amount of reading for the critical appraisal section is relatively short it is essential to practice appraising papers and those previously used are most appropriate for this. Since the examination is new advice can be difficult to obtain. An FFAEM course is immensely helpful as it allows the opportunity to ask questions of those who have recently passed the examination. PMID- 12743934 TI - Prepare for an SpR interview. AB - By the time you attend an interview for a military SpR number you should have no real problems but it pays to be prepared. Begin preparations early, reading widely and talk to as many people as possible. Your consultants will have a useful viewpoint on the proceedings and may be able to help you refine your answers to the common questions. Arrive at your interview in a smart and timely fashion and answer questions with confidence and common sense. Avoid confrontation and bluff and be courteous at all times, whatever you may be feeling inside and thank the interview panel as you leave. PMID- 12743935 TI - The Russian Florence Nightingale. PMID- 12743936 TI - The evolution of casualty evacuation in the British Army in the 20th century (Part 3)--1945 to present. AB - This is the third in a series of papers describing the evolution of the British casualty evacuation chain during the 20th century. This period was dominated by the threat of war between NATO and the Warsaw Pact in Central Europe. The Suez Crisis in 1956, the Falklands War in 1982, the GulfWar in 1991 and events in the Balkans during the 1990s demonstrated the requirement for a flexible system for medical support to the UK Armed Forces. PMID- 12743938 TI - Deciphering the "genesis problem": on the dialectical origins of psychic reality. PMID- 12743939 TI - What keeps women from reaching their goals? PMID- 12743940 TI - The changing place of the dream in psychoanalytic history, Part I: Freud, ego psychology, and the interpersonal school. PMID- 12743941 TI - Melancholic secrets: gender ambivalence and the unheimlich. PMID- 12743942 TI - Liberation discourse: a psychoanalysis of prison captivity. PMID- 12743943 TI - Freud and the suicide of Pauline Silberstein. PMID- 12743944 TI - 2003 health care legislation: opportunities for unity. PMID- 12743945 TI - Safe staffing: a serious concern. PMID- 12743946 TI - Nurse/physician communication. PMID- 12743947 TI - 2003 legislative session set to open March 4, 2003. PMID- 12743948 TI - Nurse attorney notes: think before you speak. PMID- 12743949 TI - Out of the darkness. PMID- 12743950 TI - Moving toward a tobacco-free Florida. PMID- 12743951 TI - Preparing yourself for the next decade: the GeroPsychiatric nurse practitioner. PMID- 12743952 TI - Tar wars. PMID- 12743954 TI - Unmet needs for health care. AB - OBJECTIVES: This analysis examines the prevalence of self-reported unmet needs for health care and the extent to which they were attributable to perceived problems with service availability or accessibility or acceptability. DATA SOURCE: Most data are from the 1998/99 cross-sectional household component of Statistics Canada's National Population Health Survey; 1994/95 and 1996/97 cross sectional data are used to present trends from 1994/95 to 1998/99. The primary analysis is based on 14, 143 respondents aged 18 or older. ANALYTICAL TECHNIQUES: Multivariate logistic regression was used to estimate the association of risk factors with the three types of unmet health care need. MAIN RESULTS: In 1998/99, about 7% of Canadian adults, an estimated 1.5 million, reported having had unmet health care needs in the previous year. Around half of these episodes were attributable to acceptability problems such as being too busy. In 39% of cases, service availability problems, such as long waiting times, were mentioned. Just under 13% of episodes were related to accessibility problems (cost or transportation). Unmet needs attributable to service availability problems were not significantly associated with socio-economic status. By contrast, unmet needs due to accessibility problems were inversely associated with household income. PMID- 12743953 TI - Suicide deaths and suicide attempts. AB - OBJECTIVES: This article examines suicide deaths among Canadians aged 10 or older between 1979 and 1998. It also examines hospital records for 1998/99 to provide some insight into suicide attempts. DATA SOURCES: Data are mainly from the Vital Statistics Database, the Hospital Morbidity Database, and the Person-oriented Information Database. Supplementary data are from the Adult Correctional Services and Homicide surveys, the National Longitudinal Survey of Children and Youth, and the World Health Organization. ANALYTICAL TECHNIQUES: Age-standardized rates for suicide deaths and hospitalized suicide attempts were calculated by sex and province/territory for Canadians aged 10 or older. Age- and sex-specific rates for suicide deaths and parasuicide-related hospitalizations were also calculated for seven age groups. MAIN RESULTS: The suicide death rate remained fairly stable between 1979 and 1998. The suicide rate of males was four times that of females, but females were hospitalized for attempted suicide at about one and a half times the rate of males. In 1998/99, about 9% of individuals who were hospitalized for an attempted suicide had been discharged more than once for a suicide attempt the same year. PMID- 12743955 TI - Revascularization and heart attack outcomes. AB - OBJECTIVES: This article focuses on rates of revascularization and mortality among people admitted to hospital after an acute myocardial infarction (AMI). DATA SOURCE: The hospital data are from the Person-oriented information Database. Information on deaths is from the Canadian Mortality Database. ANALYTICAL TECHNIQUES: Hospital records for Nova Scotia, Saskatchewan, Alberta and British Columbia were linked to identify AMI patients admitted between April 1, 1995 and March 31, 1996. Patients with no admission for AMI in the previous 12 months were followed for one year to determine what percentage underwent percutaneous transluminal coronary angioplasty and/or coronary artery bypass graft surgery. The risk of being revascularized and the risk of dying were estimated. MAIN RESULTS: In the year after hospitalization, 25% of AMI patients were revascularized. Rates of revascularization were relatively low for women, very elderly people, and individuals with other health problems. Revascularization was significantly associated with a lower risk of dying for male, but not female, AMI patients. PMID- 12743956 TI - Canadian community health survey--methodological overview. AB - OBJECTIVES: This article describes the design, sampling strategy, interviewing procedures, data collection and processing of the Canadian Community Health Survey (CCHS). SUMMARY: Data collection for cycle 1.1 of the CCHS began in September 2000. This first cycle provides cross-sectional data at the regional level for 136 health regions; the first half of data collected for cycle 1.1 provides data for 133 health regions. In addition to the survey methods, this article reports the sample size and rates of proxy response and non-response for each province, for the first six months of cycle 1.1. A summary of methods used to impute values that were not provided by proxy respondents is provided. A discussion of survey errors and their sources follows. PMID- 12743957 TI - Changes in unmet health care needs. AB - OBJECTIVES: This article examines recent trends in self-reported unmet health care needs among the household population aged 12 or older, and explores various explanations for the increase observed. DATA SOURCES: The data are from the first half (September 2000 through February 2001) of data collection for cycle 1.1 of the Canadian Community Health Survey and from cross-sectional (1994/95 through 1998/99) household components of the National Population Health Survey. ANALYTICAL TECHNIQUES: Weighted frequencies and cross-tabulations were used to estimate the proportion of people aged 12 or older who reported that they did not receive health care when they thought they needed it. Estimates were also produced for the type of care sought, and specific reasons for unmet health care needs. MAIN RESULTS: The percentage of people reporting unmet health care needs rose gradually between 1994/95 and 1998/99, then doubled (from 6% to over 12%) between 1998/99 and 2000/01. Long waiting time was the reason most frequently reported for unmet needs. PMID- 12743959 TI - Community belonging and health. AB - OBJECTIVES: This article explores the relationship between sense of community belonging and self-perceived health. DATA SOURCE: The data are from the first half of cycle 1.1 of the Canadian Community Health Survey (CCHS), collected from September 2000 through February 2001. ANALYTICAL TECHNIQUES: Descriptive information relating socio-demographic variables to sense of community belonging is presented. Multiple logistic regression is used to study the association between sense of community belonging and self-perceived health, while controlling for socio-demographic conditions and other health-related factors. MAIN RESULTS: Just over half (56%) of Canadians report a strong or somewhat strong sense of belonging to their local community. Community belonging is associated with self perceived health, even when controlling for socio-economic status, the presence of chronic disease, health behaviours, stress and other factors. PMID- 12743958 TI - Fruit and vegetable consumption. AB - OBJECTIVES: This article focuses on associations between the frequency of fruit and vegetable consumption and other health-related behaviours or conditions, including physical activity, smoking, obesity and alcohol-dependence. DATA SOURCE: The data are from the first half of cycle 1.1 of the Canadian Community Health Survey, collected from September 2000 through February 2001. ANALYTICAL TECHNIQUES: Weighted means provide information on average frequency of fruit and vegetable consumption in relation to selected health behaviours and conditions, health status and socio-demographic characteristics. Multivariate linear regression is used to model the associations between eating fruit and vegetables and health behaviours, while controlling for other influences. MAIN RESULTS: Women consume fruit and vegetables more often than do men. When other influences are taken into account, the frequency of eating fruits and vegetables is positively related in both sexes to being physically active, not smoking and not being overweight, and in women, to not being alcohol-dependent. PMID- 12743960 TI - In harm's way. The legacy of wartime nursing. PMID- 12743961 TI - Coping with war and terrorism anxiety. PMID- 12743962 TI - A new revolution in health care. Mental health care of older adults is vital. PMID- 12743963 TI - Positive attitude. An essential element for effective care of people with mental illnesses. AB - It is well known that attitudes influence behavior, and attitudes toward people with mental illnesses have been described as negative, stigmatizing, uninformed, and fearful. The general public view people with mental illnesses as dangerous, prone to violence, unpredictable, and in some measure responsible for their illnesses. Health care professionals share these attitudes to a surprising extent, which compromises their ability to deliver competent, compassionate care. Nursing students' perceptions of people with mental illnesses are reflective of those held by the general public. The purpose of this descriptive study was to determine whether a curriculum that embeds psychiatric nursing principles in a course focused on care of people with chronic illnesses and uses specific fear reduction teaching strategies, would have a positive effect on students' attitudes toward people with mental illnesses. Using an adaptation of Yucker's attitude scale, a test of nursing students' (n = 38) attitudes before and after completion of the course was conducted. Students' attitudes were significantly more positive at the completion of the course. The findings suggest that exposure to factual information about mental illnesses, the opportunity to interact with people with mental illnesses, emphasis on treatment options, and use of fear reduction strategies positively affected students' attitudes toward people with mental illnesses. PMID- 12743964 TI - Women's adjustment to widowhood. Theory, research, and interventions. AB - 1. People respond differently to loss and grieve in their own time. 2. Frequently, the hardest time for new widows is after the funeral. 3. Young widows often have no peer group and generally are less prepared emotionally and practically than older widows to cope with the loss. 4. Widowhood often causes financial stress because a major income source is lost with the death of a husband. PMID- 12743965 TI - Promoting self-management of urinary incontinence in a geropsychiatric day treatment program. AB - As the number of older adults in the United States increases, the number of older adults with mental illnesses also will increase. There will be a corresponding increase in prevalence of UI and its associated problems--medical problems, loss of independence or need for institutionalized care, diminished quality of life, and increased costs. Psychiatric nurses are in a position to help older adults with mental illnesses improve their overall health and quality of life by preventing the problems associated with untreated UI. Within their practice, psychiatric nurses have the opportunity to ensure clients receive the comprehensive assessments needed to establish their functional, physical, behavioral, emotional, and social support status--information that forms the foundation for developing individualized treatment interventions. Psychiatric nurses have the expertise to integrate physical and mental health care for older adults with mental illnesses and co-occurring conditions, such as UI. Promoting self-management of UI among older adults with mental illnesses potentially will enable them to participate in psychiatric rehabilitation programs; improve their overall health and quality of life; prevent falls and fractures that often cause them to lose their independent community living status and to be admitted to long term care facilities; and reduce the cost to mental health care providers of managing UI in the treatment setting. PMID- 12743966 TI - Finding effective strategies for working with adolescents. PMID- 12743967 TI - The myths of drug addiction. PMID- 12743968 TI - A collegial mentoring model for nurse educators. AB - TOPIC: A mentoring model for nurse educators. PURPOSE: To describe the current literature on mentoring in nursing and education and present the Collegial Mentoring Model. SOURCES: Published literature and personal and professional experiences of nurse educators in academia. CONCLUSIONS: Commitment to the collegial mentoring relationship can be demonstrated by making time for togetherness so outcomes beneficial to both individuals can be achieved. PMID- 12743969 TI - Cost issues related to American healthcare policy. AB - TOPIC: Diverse issues that affect current and future healthcare costs. PURPOSE: To review the historical, present, and future cost issues related to American healthcare policy. SOURCES: Nursing journals, public policy Web sites, and health policy literature. CONCLUSIONS: Any real change in health care will be hard to make because it always comes down to "Who pays?" Productive and sensible policy reform is preferable to the risk we now run of a healthcare system disconnected from any values except those of the marketplace. PMID- 12743970 TI - A look at nursing leadership through the lens of a dancer. AB - TOPIC: Using the world of dance to describe leadership dimensions. PURPOSE: To explore the art of nursing leadership from the perspective of dance, a form of artistry through movement. SOURCES: Published literature and the author's own experience. CONCLUSIONS: The metaphors of dance bring the concept to life in a new way. PMID- 12743971 TI - Computers: do they help or hinder patient care? PMID- 12743972 TI - A conversation with the family caretaker of a dying man. PMID- 12743973 TI - Social justice, human rights, and nursing education. PMID- 12743974 TI - Environmental education and practice guidelines. PMID- 12743976 TI - Preparing nurses for a 21st century role in genomics-based health care. AB - Advances in genomic research are changing the nature and focus of health care. All health professionals must be prepared with new knowledge and skill competencies to meet the social and scientific demands of clinical practice. Nurses can expect to take on new practice roles that involve family history assessments, screening, and case coordination for clients receiving genetic testing and gene-based therapies. The authors report on successful initiatives since 1995 resulting from efforts of national organizations to advance the diffusion of genetic knowledge and genomics in nursing education and practice. Recommendations to strengthen these developments, along with strategies for nursing leadership in carving a new role, that of Genomics Nurse Case Coordinator, are presented. PMID- 12743975 TI - Technology-based vs. traditional instruction. A comparison of two methods for teaching the skill of performing a 12-lead ECG. AB - The purpose of this study was to compare the effectiveness of an interactive, multimedia CD-ROM with traditional methods of teaching the skill of performing a 12-lead ECG. A randomized pre/posttest experimental design was used. Seventy seven baccalaureate nursing students in a required, senior-level critical-care course at a large midwestern university were recruited for the study. Two teaching methods were compared. The traditional method included a self-study module, a brief lecture and demonstration by an instructor, and hands-on experience using a plastic manikin and a real 12-lead ECG machine in the learning laboratory. The second method covered the same content using an interactive, multimedia CD-ROM embedded with virtual reality and supplemented with a self study module. There were no significant (p < .05) baseline differences in pretest scores between the two groups and no significant differences by group in cognitive gains, student satisfaction with their learning method, or perception of self-efficacy in performing the skill. Overall results indicated that both groups were satisfied with their instructional method and were similar in their ability to demonstrate the skill correctly on a live, simulated patient. This evaluation study is a beginning step to assess new and potentially more cost effective teaching methods and their effects on student learning outcomes and behaviors, including the transfer of skill acquisition via a computer simulation to a real patient. PMID- 12743977 TI - Outcomes of a genetics education program for nursing faculty. AB - A multifaceted educational program designed to teach nursing faculty about genetics was first offered in 1997 and subsequently repeated on an annual basis. The specific aims of the program were to: 1) increase nursing faculty knowledge about genetics and its clinical application, and 2) increase genetics content taught in entry-level nursing education programs. The major components of the program included an annual Genetics Summer Institute (GSI), pre-planned follow-up strategies, and continuing education offerings. Measured outcomes included significant improvement in nursing faculty genetics knowledge and increased amounts of genetics content in their curricula. The majority of surveyed faculty focused curriculum change efforts on lectures or courses for which they were personally responsible. Thirty-one percent were working on, or had developed, elective nursing genetics courses after attending a GSI. These findings indicate that this program provided the necessary foundational instruction and resources to enable nursing faculty participants to bring about change in their curricula. PMID- 12743978 TI - Oncology nursing education: nursing students' commitment of "presence" with the dying patient and the family. AB - During an elective oncology nursing course, students expressed uncertainties about activities that would offer patient and family support during end-of-life care. Following a chaplain's lecture, students in a class reaction paper identified appropriate nurse responses and actions that would offer supportive care to the dying patient and the family. Six processes were extracted from student comments. A core category was identified as the importance of "nurse presence" at the bedside of the dying patient. PMID- 12743979 TI - Facilitating transformation through collaboration. AB - The philosophical underpinnings of transformative education were utilized to redesign a course taught to special education majors and nursing students at a rural southeastern university. The main goal when restructuring the class was to develop a course that would encourage communication and collaboration between education and nursing majors within the school community. The current course, "Health Care Perspectives of Exceptional Students," focuses on specific health care topics, with nurse faculty presenting the health care aspects and education faculty discussing educational implications for the classroom teacher. Every class includes dialogue and activities that require interaction. The last two classes are devoted to first-aid training and CPR certification. During the evaluation phase of the course, both education and nursing majors responded positively to the question of whether or not medically fragile children belong in the classroom. Furthermore, they saw the need for communication and collaboration between the classroom teacher and the school nurse as essential for effectively serving the medically fragile child in the school setting. PMID- 12743980 TI - The development of a community-based baccalaureate curriculum model in a culturally diverse health care delivery area. AB - In 1997, faculty at the University of Texas Health Science Center at San Antonio School of Nursing embarked on the development of a community-based care model within the existing curriculum to serve the culturally diverse populations in rural and urban areas of South Texas. A grant awarded by the Helene Fuld Health Trust in 2000 enabled faculty to develop the requisite knowledge and skills relating to community-based care and develop strategies for the delivery of community-based care in a variety of settings. This article provides an overview of the process of curriculum development and its impact on faculty, students, and a diverse community service area. PMID- 12743981 TI - Phase II trial of pegylated-liposomal doxorubicin (Doxil) in sarcoma. AB - Pegylated-liposomal doxorubicin (Doxil) is a unique form of liposomal doxorubicin in which the liposomes are coated with methoxypoly (ethylene glycol), resulting in a diminished uptake by the reticuloendothelial system, leading to a longer half-life in blood and a different toxicity profile than nonpegylated liposomes. We performed a phase II study of Doxil in sarcoma. The patient population was primarily previously treated or had diagnoses considered unresponsive to chemotherapy. The initial dose per course was 55 mg/m2 every four weeks with dose modification based on mucositis and hand-foot syndrome (the main limiting toxicities). Treatment was generally well tolerated. Of 214 evaluable treatment courses in 47 patients, toxicities were mild and similar to previous reports, but dose reduction was common. No definite cardiac toxicity was observed. There were: 6 osteosarcomas, 3 Ewings, 1 extraosseous osteosarcoma, 1 chondrosarcoma, 2 alveolar soft part sarcomas, 15 gastrointestinal stromal cell tumors (GIST), and 19 other soft tissue sarcomas. Three of the 47 patients received a CR or PR, although 15 of the 47 patients were felt to have derived clinical benefit from the treatment. Some responses were delayed. These data suggest that pegylated liposomal doxorubicin has activity in this population of poor prognosis sarcoma and that this treatment is associated with modest toxicity. PMID- 12743982 TI - A pilot study of edrecolomab (Panorex, 17-1A antibody) and capecitabine in patients with advanced or metastatic adenocarcinoma. AB - Edrecolomab (Panorex) is a monoclonal antibody directed against the 17-1A antigen located on the cell surfaces of carcinomas. Clinical activity has been seen in colon and breast cancer. This trial investigated the feasibility of combining edrecolomab with the oral fluoropyrimidine capecitabine (Xeloda). Patients received a loading dose of edrecolomab 500 mg intravenously (i.v.) on day--14, followed 2 weeks later by 100 mg i.v. every 28 days (day 1). Capecitabine was administered to single-patient cohorts at escalating doses of 1500, 2000, and 2500 mg/m2/day in two equally divided doses for 14 of 21 days, beginning on day 1. Additional patients were enrolled at the 2500 mg/m2/day dose level to better define the toxicities of combination therapy. Toxicity assessment was the primary endpoint. Twenty seven patients with advanced or metastatic adenocarcinoma were enrolled on this study: 20 were evaluable for toxicity and 18 for response. The most common toxicities were elevated liver enzymes, diarrhea, and hand-foot syndrome. In cycle 1, grade 3 hand-foot syndrome was seen in two patients, and grade 3 diarrhea in one patient. Grade 2 toxicities included diarrhea, hand-foot syndrome, anemia, leukopenia, and transaminitis. Cumulative hand-foot syndrome was observed in four patients treated beyond two cycles. Three patients had edrecolomab infusion reactions during the course of treatment. One complete response and two partial responses were seen. Nine patients had disease stabilization lasting a median of 17.5 weeks (range 14.5-28+). Edrecolomab and capecitabine may be safely given in combination to patients with advanced or metastatic adenocarcinoma. Clinical activity is seen in this heavily pretreated patient population. PMID- 12743983 TI - Effect of an intensive chemotherapy followed by mediastinal irradiation on pulmonary and cardiac function in advanced Hodgkin's disease. AB - Mediastinal irradiation combined with chemotherapy in patients with Hodgkin's disease have been associated with cardiopulmonary toxic effects that can last over the years. In this study we monitored pulmonary and cardiac function in 39 patients affected by advanced Hodgkin's disease (stage II B-III and IV) with mediastinal involvement and submitted to an intensive chemotherapy regimen (epirubicin, vincristine, ciclophosphamide, and etoposide) followed by involved field irradiation. Pulmonary function was verified with chest x-ray, spirometric parameters, arterial blood gas analysis, single breath CO transfer factor (DLCO), and its components Dm and Vc. Cardiac function was verified with electrocardiogram (EKG) and left ventricular ejection fraction (LVEF) by means of radionuclide angiocardiography. The median follow-up was 40 months. Spirometric parameters did no show modifications at the end of treatment, on the contrary they improved during the follow-up. Chest x-ray showed radiographic parenchimal damage in 51% of patients. DLCO remained constantly decreased. sEKG did not show significant modification, whereas LVEF significantly decreased at the end of treatment and remained persistently decreased during follow-up. None of the patients with reduction of DLCO or LVEF showed clinical symptoms of heart and pulmonary dysfunctions. One patient, 49 years old, suffered from myocardial infarction 25 months after the completion of radio-chemotherapy. These data indicate that this combined regimen can induce persistent pulmonary and cardiac damages at subclinical level. PMID- 12743984 TI - Combination chemotherapy with gemcitabine and vinorelbine in the treatment of patients with relapsed or refractory small cell lung cancer: a phase II trial of the Minnie Pearl Cancer Research Network. AB - The purpose of the study was to evaluate the combination of gemcitabine and vinorelbine in the treatment of patients with relapsed or refractory small cell lung cancer. Between March 1998 and February 1999, 30 patients with relapsed or refractory small cell lung cancer who had received treatment with one previous combination chemotherapy regimen entered this multicenter, community-based clinical trial. All patients had received previous platinum/etoposide combination chemotherapy; in addition, 12 patients had received paclitaxel as part of their first-line therapy. All patients received gemcitabine 1000 mg/m2 and vinorelbine 20 mg/m2 on days 1, 8, and 15 of each 28-day cycle. Patients were reevaluated for response after two cycles of therapy; those with objective response or stable disease continued treatment for six courses or until disease progression. Three of 28 evaluable patients (10%) had partial responses to treatment. None of the 17 patients with refractory disease responded, while 3 of 12 patients (25%) with relapsed disease had partial responses. Median survival was 5 months. Treatment was generally well tolerated; myelosuppression was the major toxicity, but only two patients developed febrile neutropenia, and there were no treatment-related deaths. Non-hematologic toxicity was uncommon; alopecia did not occur with this regimen. The activity of gemcitabine and vinorelbine in patients with previously treated small cell lung cancer is modest and is limited to patients with relapsed (versus refractory) disease. In patients with relapsed small cell lung cancer, this regimen provides an additional treatment option, with decreased toxicity when compared to other second-line options. However, novel treatment approaches are necessary before substantial improvements in treating this patient population will be realized. PMID- 12743985 TI - Differential effects of cigarette smoke extracts on cell proliferation in gastric and colon cells. AB - Substantial evidence show a higher incidence of gastric cancer in smokers than nonsmokers and that cigarette smoking is highly associated with colon cancer. The present study was designed to examine the effect of cigarette smoke extracts on gastric and colon cancer cell proliferation, which is important for tumor growth. Two different cell lines were used. One was gastric cancer cell line AGS, and the other was colon cancer cell line HT-29. It was found that cigarette smoke extracts stimulated cell proliferation and c-myc expression in AGS cells. Furthermore, this proliferative action was partially blocked by the c-myc antisense. However, the extracts significantly inhibited HT-29 cell proliferation and suppressed c-myc expression. In conclusion, cigarette smoke extracts stimulated AGS cell proliferation, while inhibiting HT-29 proliferation, which were partially mediated by a c-myc-related pathway. The former action may play a contributory role in the carcinogenic action of cigarette smoking in the stomach. PMID- 12743986 TI - A high cytosol value of urokinase-type plasminogen activator (uPA) may be predictive of early relapse in primary breast cancer. AB - BACKGROUND: There is now much data that suggest a relationship between angiogenesis and breast cancer prognosis. Angiogenesis is a multistep process resulting from an ordered set of events and regulated by positive and negative modulators of microvessels growth and by the expression of various proteolytic enzymes. MATERIALS AND METHODS: We prospectively evaluated VEGF and microvessels density on tumor specimen and cytosolic levels of uPA and PAI-1. RESULTS: We enrolled 81 primary breast cancer patients. The median follow-up was 38 months. Using the median value as cutoff for the statistical analysis, we found significant correlation between cytosolic levels of uPA and PAI-1 (r = 0.61; p < .0001), between VEGF and steroid hormone receptor status (p = .01), between PAI-1 and tumor grading (p = .009), and between uPA and tumor size greater than 1 cm (p = .04). With respect to the prognosis, we observed a significant correlation between low uPA levels and RFS and an unforeseen, direct correlation between high VEGF values and better RFS. CONCLUSIONS: Our preliminary results indicate that the cytosolic level of uPA at diagnosis may be predictive of early relapse in primary breast cancer. PMID- 12743987 TI - Synergistic effect of sequential administration of mitoguazone (MGBG) and gemcitabine in treating tissue cultured human breast cancer cells and mammary rat tumors. AB - Modulation of cancer chemotherapeutic drugs has been attempted to increase efficacy and overcome resistance to the chemotherapeutic agent. Studies have shown schedule-dependent interactions in combined use of chemotherapeutic drugs. Mitoguazone (MGBG), an old drug with possible modulating activity, was used in combination with gemcitabine, a relatively new cancer drug, in treating tissue cultured human breast cancer cells and mammary rat tumors. Tissue cultured BOT-2 cancer cells were first treated with varying concentrations of gemcitabine and MGBG, independently. Combinations of the two drugs were then used with different scheduled administrations. Marked synergistic activity was found between gemcitabine and MGBG when the MGBG was given first, followed by gemcitabine 24 hours later. A non-toxic dose of MGBG enhanced the toxicity of gemcitabine by eight orders of magnitude using MTT assays in the tissue cultured human breast cancer cell study. The sequential administration of MGBG and gemcitabine also increased the survival rate of rats bearing mammary tumors in our pilot animal study. PMID- 12743988 TI - Tumor-host interactions accompanying the growth of the G:5:113 fibrosarcoma in the mouse: possibilities for a new therapeutic approach? AB - The experiments were aimed at describing in detail some interactions between a solid tumor growing from subcutaneously transplanted G:5:113 fibrosarcoma cells in vivo and its mouse host. The tumor was found to elevate significantly the number of granulocytes in the peripheral blood of the host after having achieved the volume of about 1 cm3 (day 40 after transplantation). Blood plasma from fibrosarcoma-bearing mice stimulated proliferation of progenitor cells for granulocytes and macrophages (GM-CFC) in vitro and suppressed growth of G:5:113 cell population in culture. Interestingly, both effects were observable as early as week 1 when the tumor was still macroscopically invisible and unpalpable. Conditioned medium from cultures of G:5:113 fibrosarcoma cells stimulated proliferation of GM-CFC in vitro. These findings might represent a starting point for studies aimed at designing new therapeutic approaches for the treatment of fibrosarcoma. PMID- 12743989 TI - Food groups and risk of esophageal cancer in Chaoshan region of China: a high risk area of esophageal cancer. AB - The current study was designed to investigate the role of common foods in the etiology of esophageal cancer in the Chaoshan region of China. A large case control study was conducted to investigate 1248 patients with squamous cell carcinoma of the esophagus and 1248 controls matched by sex, age, and hospitals. After adjusting for the effects of sex, age, occupation, areas of residence, income, alcohol intake, and cigarette usage, a strong association with a clear dose-response relationship was observed between the fermented fish sauce eaten weekly and esophageal cancer (P for trend less than 0.001). The results for pickles were similar to those for fermented fish sauce. A protective effect of fruits on esophageal cancer was observed, and this risk decreased (P for trend less than 0.001) as frequencies of fresh fruit intake increased. The risks from sowbelly and kipper were significantly higher in consumers than in nonconsumer, but the dose-response relationship was not statistically significant. No significant effects, however, were detected from vegetables, smoked foods, and so on. To the authors' knowledge, this is the first report on the relationship between fish juice and high risk of esophageal cancer in a population. Further epidemiological and experimental study are required to find a biological causal relationship between them. PMID- 12743990 TI - Comparative physical and pharmacologic characteristics of iodine-131 and yttrium 90: implications for radioimmunotherapy for patients with non-Hodgkin's lymphoma. AB - Radioimmunotherapy (RIT) is a promising new treatment option for patients with relapsed/refractory non-Hodgkin's lymphoma. Clinical trials have demonstrated that both iodine-131 (131I) and yttrium-90 (90Y) are suitable radionuclides for RIT. Iodine-131 and 90Y differ markedly in their physical properties including half-life, path length, type of energy emissions, intracellular stability, and the organs targeted by the free radionuclide. Both radionuclides can be safely administered in the outpatient setting under current Nuclear Regulatory Commission guidelines. Potential advantages of 131I for RIT include availability, stable chemistry, longer half-life, and an emission spectrum that allows for dosimetric studies and therapy with the same immunoconjugate. By contrast, 90Y has a longer path length and superior intracellular stability compared with 131I. Yttrium-90 may therefore be preferable to 131I for patients with bulky disease, poorly vascularized solid tumors, or when targeting internalized antigens. Although 90Y emits no gamma photon, dosimetry studies for 90Y RIT can be performed using a surrogate radionuclide such as indium-111. Both 131I- and 90Y labeled anti-CD20 antibodies have demonstrated efficacy in treating relapsed/refractory non-Hodgkin's lymphoma. Further studies are needed, however, to determine if the differences in the pharmacology of 131I and 90Y are clinically relevant. PMID- 12743991 TI - The human T-cell lymphoma/leukemia viruses. AB - The primate T-cell lymphoma/leukemia viruses belong to an oncogenic genus of complex retroviruses. Members of this genus have been shown to be pathogenic in man. The human T-cell lymphoma/leukemia virus (HTLV) type I has been linked in the etiology of T-cell malignancies and "autoimmune-like" neurologic and rheumatic disorders; a related virus, HTLV-II, is becoming increasingly associated with similar disorders. Cell transformation is thought to be caused predominantly by the effects of the viral regulatory protein, Tax. An additional induced host cell molecule, adult T-cell lymphoma-derived factor, may contribute to cell immortalization. Like the DNA tumor viruses, HTLV activates transcription of cellular proto-oncogenes and inhibits cellular mechanisms of tumor suppression, cell cycle arrest, and apoptosis. However, individuals who are able to mount a strong cell-mediated immune response and limit viral entry into uninfected cells do not develop associated malignancies. Unfortunately, HTLV induced malignancies are difficult to treat with conventional chemotherapy, and disease progression is often rapid with a median survival of less than 2 years. There are, however, some novel approaches that have yet to be fully tested that may have greater efficacy in the treatment of HTLV-induced diseases. In the future, better screening and detection methods, along with new vaccines and therapies, may contribute to the increased prevention and control of HTLV infection and its associated diseases. PMID- 12743992 TI - Genetically engineered herpes simplex viral vectors in the treatment of brain tumors: a review. PMID- 12743993 TI - Molecular phenotyping of the immune system by microarray analysis. PMID- 12743994 TI - Retinoids, retinoic acid receptors, and breast cancer. AB - Retinoids comprise both naturally and synthetically occurring compounds that have been proven to be differentiation agents for a variety of neoplasias, including breast cancer and promyelocytic leukemia in animal models and humans. They offer a unique panoply of therapeutics for the prevention or treatment of breast cancer. Nonetheless, considerable controversy remains as to the efficacy and potential toxic side-effects and as to which group of patients may most benefit. In this article, we review evidence of retinoid efficacy in breast cancer in humans and in animal models and provide possible mechanisms of retinoid action in breast cancer treatment, focusing on the roles of the different retinoic acid receptors and the metabolic pathways necessary for gene activation and cellular homeostasis. PMID- 12743995 TI - Establishing a palliative care program in a research center: evolution of a model. PMID- 12743996 TI - Treatment of sarcomas: thinking out-of-the-box. PMID- 12743997 TI - Leukemia and myelopathy: the persistent mystery of pathogenesis by HTLV-I/II. PMID- 12743998 TI - The Cancer Genome Anatomy Project: power tools for cancer biologists. PMID- 12743999 TI - Retinoid signaling in breast carcinoma cells. PMID- 12744000 TI - Great expectations. PMID- 12744004 TI - A case of suicide disguised as natural death. AB - A case report describing the suicide of a physician classified initially as a natural death. Faced with the vehement protest of the family of the deceased the magistrate decided to request a 'preventive' forensic autopsy. Forensic investigations revealed the cause of death as being a pentobarbital intoxication and the circumstances favoured the hypothesis of a genuine suicide. This case illustrates that the tendency of magistrates to request or not an autopsy is related to their experience or intuition. PMID- 12744005 TI - The identification of a US serviceman recovered from the Holy Loch, Scotland. AB - During February and March 2000, human remains were recovered from the Holy Loch, Scotland. Police enquiries identified 13 males that had gone missing, presumed drowned in the Holy Loch or the adjoining lochs, over the previous 35 years. Osteological examination of the remains established they were from a male, aged between 15 and 23 and 168-174 cm tall. This information eliminated ten of the known missing persons. DNA profiles, both STR and mitochondrial were generated from the remains and compared to the profiles generated from relatives of the missing men. A positive match between the unidentified individual and one of the maternal relatives identified the remains as belonging to a US serviceman who had gone missing 35 years ago. The successful identification led to the repatriation of the serviceman's remains. PMID- 12744008 TI - [The supplement on 100 years' of ECG--genesis of its inception]. PMID- 12744007 TI - Forensic science coming of age. PMID- 12744009 TI - [One hundred years' of the electrocardiogram--a look at the past and view of the future ]. PMID- 12744010 TI - [Pathways in the development of electrocardiography]. AB - Electrocardiography celebrates hundred years of its existence. The historical article on the cornerstones of its development was written on request of the editorial board of Vnitrni lekarstvi (Internal Medicine) and the Czech Society of Internal Medicine. After summarizing the beginnings of the discovery of bioelectric current and attempts of its registration the author describes the construction of the string galvanometer by Willem Einthoven. In 1901 and 1903 he published the first findings on the application of electrocardiography in man. Therefore that years are considered the years when clinical electrocardiography was born. Next we deal with the development of lead systems and modern electrocardiographs, incl. the discovery of the Braun tube, the construction of monitors, vectorcardiography and spatiocardiography. The author summarizes the principles of rational interpretation of the surface electrocardiogram and analyzes further modifications such as the use of leads from the right hemithorax, oesophageal leads, stress electrocardiography. Classical surface electrocardiography is probably a completed discipline. It will remain also in future a basic auxiliary examination method in internal medicine and allied disciplines. Due to its importance however classical electrocardiographic examination should be shifted into primary care where it may be a great enrichment and acceleration of the diagnostic process. Further development of electrocardiology is focused on non-invasive and invasive examinations and in particular treatment of arrhythmias where surface electrocardiography will be also in future important as a reference signal for the time localization of electric potentials of the intracardial electrocardiogram and programmed electrical stimulation. PMID- 12744011 TI - [The beginnings of electrocardiography at the Prague Medical School]. AB - The authors describe the beginnings of development of electrocardiography at the Prague Medical Faculty of Charles-Ferdinand and Charles University resp. The first results of the new method came from the physiological laboratory of Ewald Karl Konstantin Hering (1834-1918), who contributed to electrocardiography by explaining the mechanism of atrial fibrillation and to Richard Hans Kahn (1876 1941) who was the first to publish electrocardiographic changes after temporary block of coronary artery blood flow in the dog. Clinical application and the promotion of electrocardiography in this country is among others in particular from Vaclav Libensky (1877-1938), Klement Weber (1890-1971) and Frantisek Herles (1900-1991) whose name became for half a century the synonym for electrocardiography. PMID- 12744012 TI - Willem Einthoven--inventor of electrocardiography. AB - Willem Einthoven, the inventor of the string galvanometer electrocardiograph, was born in the Dutch East Indies, studied medicine in Utrecht Holland, and became chairman of the Department of Physiology at the University of Leiden. In 1924 he was awarded the Nobel Price for physiology and medicine. Einthoven became interested in electrophysiology after Waller's demonstration of the human electrogram using Lippmann's capillary electrometer. Because of important limitations of this device, Einthoven decided to construct the string galvanometer to be used for physiological research and in clinical medicine. Einthoven's main achievements in electrophysiology were the description of the normal ECG, some physiological effects on the ECG, the three ECG leads and the triangle rule. In 1906 in cooperation with the university hospital, Einthoven demonstrated the clinical usefulness of the electrocardiograph. PMID- 12744013 TI - A history of atrial fibrillation. AB - Vulpian in France and Hoff and Ludwing in Germany first studied atrial fibrillation in experimental animals. In clinical medicine, de Senac and others, first observed irregular and fast pulse. Hering in 1903 described "pulsus irregularis perpetuus" as a distinct arrhythmia. Hering believed, that this was an extrasystolic rhythm disturbance, while Cushny and Edmunds suggested atrial fibrillation as its cause. Mackenzie was another British author who played an important role in the clarification of atrial fibrillation, particularly after Cushny convinced him, that the so-called nodal rhythm was atrial fibrillation. Einthoven reported the first ECG of atrial fibrillation without recognizing its true nature. Finally in 1909 and 1910 Lewis in London and, Rothberger and Winterberg in Vienna established atrial fibrillation as a clinical entity. PMID- 12744014 TI - [100 years' of ECG or 100 years' of excellence in recording the electrical activity of the human heart]. AB - The authors present a brief history of high-standard records of the electric activity of the human heart. Its beginnings date back to 1892 when Willem Einthoven recorded the first electrocardiogram. PMID- 12744015 TI - [Perspectives in electrocardiography 100 years after Einthoven's discovery. Personal views]. PMID- 12744016 TI - [The electrocardiogram and thrombolytic therapy in patients with acute myocardial infarct]. AB - The objective of the submitted work is to analyze in patients with acute myocardial infarction (AIM) local priority data on ECG markers after admission to hospital, data on some associations of ECG and thrombolytic treatment and to assess in patients with the first AMI data on hospital mortality in connection with some ECG markers. The project was implemented as a prospective multicentre study. An independent audit and collection of data was done in 3123 patients with AIM in 66 departments between Sept. 16 1997 and Sept. 15 1998. The group included patients admitted within 96 hours after development of complaints with the diagnosis or suspicion of AMI who were discharged with the diagnosis of a first/repeated AMI. Elevation of ST segments was recorded in 67.1%, a Q wave in 42.2% and left bundle branch block in 3.7% of the patients. Early diagnosis of AMI based on ECG and data on prolonged stenocardia was made in 55.6% patients. This is the maximal proportion of patients where thrombolytic treatment can be contemplated. Thrombolytic treatment was not administered to 54.9% patients with elevations of the ST segments and in as many as 81.2% patients with left bundle branch block. The hospital mortality in patients with a first AMI is significantly greater in patients with elevations of the ST segment, Q infarction, anterior wall infarction, combined infarction, right ventricular infarction and in patients with bundle branch and fascicular block. It was confirmed that in Slovakia in clinical practice thrombolytic treatment is not always administered consistent with criteria adopted from randomized studies. The result is underutilization or overutilization of thrombolytic treatment to patients with AIM in clinical practice. Underutilization of thrombolytic treatment is generally known. It was demonstrated that attention must be devoted also to overutilization of thrombolytic treatment. All patients where significantly higher hospital mortality was recorded must receive special care already on admission to hospital. PMID- 12744017 TI - [The ECG in prediction of the infarcted artery and left ventricular dysfunction]. AB - OBJECTIVE: To assess the possibilities of ECG in prediction of an infarcted artery in patients with acute myocardial infarction (AIM) and evaluate the relationship of left ventricular function to the number of pathological Qs. METHOD: The trial comprises 216 patients with acute myocardial infarction (AMI) treated by primary PTCA. The diagnosis of AIM was confirmed in addition to ECG also by coronarography and elevation of cardiospecific enzymes. On the baseline 12-lead ECG denivelization of the ST segment > 1 mm was considered significant. The number of pathological Q waves on the ECG tracing on discharge was compared with left ventricular function according to the ejection fraction assessed echocardiographically before discharge. RESULTS: Elevation of ST in leads V1-V4 predicts occlusion of LAD with a sensitivity of 96% and specificity of 69% (p < 0.001 vs. RC segment with a sensitivity of 80% and specificity of 48% (p < 0.001). In inferior AMI the ST elevation in III > II develops in 70% in occlusion of RCA vs. 27% in occlusion of LCX. The mean left ventricular ejection fraction before discharge was in patients with four or fewer pathological Q waves 49 +/- 10% vs. 32 +/- 9% in the presence of pathological Q waves in > 4 leads. CONCLUSION: In patients with AMI according to ECG changes in different leads conclusions may be drawn on the infarcted artery. The presence of pathological Q waves in more than 4 leads predicts severe left ventricular dysfunction. PMID- 12744019 TI - [Role of electrocardiography in the diagnosis of silent myocardial ischemia]. AB - Norman J. Holter attention drew to the possibility to use ambulatory ECG monitoring not only for the diagnosis of arrhythmias but also myocardial ischaemia by already in 1961. It was found that patients with chronic stable angina pectoris have multiple episodes of myocardial ischaemia characterized by transient depressions of the ST segment, very frequently without a clinical correlate. Electrocardiography thus was the first objective method to draw attention to the fact that angina pectoris is associated with approximately one fifth of ischemic episodes of the myocardium in patients with chronic ischaemic heart disease. This was later expressed objectively by further methods which with a different sensitivity and specificity can diagnose current myocardial ischaemia. It was found that developing myocardial ischaemia is gradually manifested within several to tens of seconds by detectable metabolic changes (PET, sampling from the coronary sinus), changes at the level of the microcirculation with impaired perfusion (thalium scan, contrast echocardiography, PET, contrast angiography), impaired diastolic function of the left ventricle (direct assessment of pressures, Doppler echocardiography), regional disorders of left ventricular kinetics (contrast ventriculography, echocardiography, isotope ventriculography). Only after that electrocardiographic signs of myocardial ischaemia develop, in some patients associated with angina pectoris. It is obvious that the diagnosis of myocardial ischaemia by electrocardiographic signs and subjective manifestations is late and not very sensitive. PMID- 12744018 TI - [Risk stratification in unstable angina pectoris and non-Q myocardial infarct: anamnesis, physical examination, electrocardiography, specific cardiac markers]. AB - Patients with unstable angina pectoris or non Q-infarction of the myocardium are a very heterogeneous group. It comprises patients with angina pectoris which developed for the first time in their life and had only an episodic character, as well as patients with a severe clinical course and prognostically serious coronary finding. In this connection risk stratification of patients is useful as a provision from which the further diagnostic and therapeutic procedure develops. Based on an analysis of anamnestic data, physical examination, electrocardiogram and examination of cardiospecific, biochemical markers we can divide the patients into groups with a high, medium and low risk. In patients with a high risk the authors use in addition to standard medicamentous treatment also an early invasive examination with possible revascularization. Coronary angiography without the necessity of immediate implementation is also indicated in the majority of patients with medium risk. Conversely low risk patients do not require hospital admission in some cases, or they may be discharged soon. PMID- 12744020 TI - [What does the ECG reveal in infectious endocarditis?]. AB - The electrocardiogram should be recorded at the very beginning of infectious endocarditis. It provides information on the origin and complications of possible organic disease (aortal stenosis--left ventricular hypertrophy). Moreover regular follow up of ECG tracings can reveal changes signalizing spread of the infection beyond the endocardium. Thus the conduction system may be affected--bundle branch block, atrioventricular blocks grade I to III, ectopic functional tachycardia, extrasystoles, myocardium--by development of typical ECG changes during myocardial infarction after embolization of the vegetation into the coronary artery, or last not least, the pericardium, after spread of the infection into the pericardial cavity with the serious finding of purulent pericarditis with diffuse ST-T elevations on the ECG tracing. PMID- 12744021 TI - [The importance of electrocardiography in cardiomyopathies]. AB - The author discusses possibilities and the importance of electrocardiography in different types of cardiomyopathies. ECG plays a fundamental role in particular in hypertrophic cardiomyopathies where it serves for screening. This pertains to fully developed forms as well as to latent morphologically not yet expressed forms and also to some special types of the disease (apical forms). ECG can be a very important method also in right ventricular arrhythmogenic cardiomyopathies (again above all for screening in paroxysmal ventricular tachycardias in left bundle branch block). In other types of cardiomyopathies the importance of ECG is small. PMID- 12744022 TI - [Electrocardiography after heart transplantation]. AB - In the submitted review the authors describe the position and importance of electrocardiographic examination after transplantation of the heart (HTx). They describe findings recorded in animal experiments, typical ECG characteristics after HTx and their importance as well as the clinical application of this method for diagnosis of electrophysiological abnormalities specially in relation to rejection and reinnervation of the graft. The authors' conclusions are based on a critical analysis of relevant data in the literature as well as their own experimental and clinical experience. PMID- 12744023 TI - [Electrocardiographic changes after heart transplantation]. AB - Electrocardiographic (ECG) changes are described after heart transplantation in almost 75% patients. During the early postoperative period the usual finding are conduction disorders which in 3-5% call for implantation of a pacemaker. The most frequent persisting disorder is bundle branch block which is of clinical importance only when it has a progressive character. The incidence of postoperative atrial fibrillation or flutter is lower as compared with other cardiosurgical operations and their sudden development may be associated with acute rejection. Ventricular arrhythmias develop as a rule as a complication of advanced coronary disease of the graft and are frequently the cause of sudden death. Before the introduction of cyclosporin A a relatively reliable sign of acute rejection was a reduction of the QRS complex voltage. During contemporary treatment ECG changes develop only in severe forms of rejection, incipient changes can be recorded only by an intracardial electrogram. PMID- 12744024 TI - [Importance of the ECG for evaluation of severity of pulmonary embolisms]. AB - OBJECTIVE: To evaluate whether it is possible on the basis of ECG examination to judge the haemodynamic impact of pulmonary embolism. METHOD: To compare by retrospective analysis of hospital documentation of patients at the medical department the assessed ECG changes with the echocardiographic finding of pulmonary hypertension and right ventricular dilatation. As positive an ECG finding was evaluated involving changes of the ST-T segment in VI-V4, S1Q3T3 negative or right bundle branch block (RBBB). As a positive echocardiographic finding the authors evaluated the size of the right ventricle above 30 mm or the shape of the acceleration curve in the outflow tract of the right ventricle corresponding to pulmonary hypertension. RESULTS: The authors analyzed 179 patients with clinically diagnosed pulmonary embolism during the period from Jan. 1 1996 till Dec. 31 2000. A positive ECG finding was recorded in 103 patients, i.e. 57.5%. Echocardiographic examination was made in 112 and 65 of them had a positive ECG. An echocardiographic finding of pulmonary hypertension or dilatation of the right ventricle was recorded in 92 patients. In case of positivity of at least one ECG finding an ECG sensitivity of 62% and specificity of 60% was found. In case of changes in the ST-T segment in V1-V4 the sensitivity and specificity was 47% and 70% resp., for S1Q3T3 negative 27% and 80% resp. and PRT block 25% and 90% resp. CONCLUSIONS: The absence of ECG changes does not rule out serious pulmonary embolism. The finding of typical ECG changes is very frequently associated with pulmonary hypertension, right ventricular dilatation and a serious haemodynamic finding. PMID- 12744025 TI - [Importance of the ECG in pulmonary thromboembolisms in gynecology]. AB - The authors analyze the diagnostic value of ECG examination in pulmonary thromboembolism which is a frequent complication in elderly women in postoperative gynaecological departments. They draw attention in particular to the importance of ECG in the differential diagnosis between cor pulmonale acutum and infarction of the inferior and anterior myocardial wall. PMID- 12744026 TI - [ECG changes in aortic valve defects]. AB - ECG is nowadays no longer the dominant way of diagnosing aortic valve diseases. The basis of accurate diagnosis of these diseases is clinical examination, ECHO and catheterization of the heart, which is essential in the great majority of valve diseases, in particular to rule out coronary changes. ECG is however important for the primary evaluation of left ventricular hypertrophy, left ventricular overburdening, for detection of enlargement or overloading of the left atrium. It is irreplaceable for evaluation of impaired rhythm, or impaired conduction of the impulse. Thus we can prove atrial fibrillation, or flutter bundle branch block, disorders of AV conduction (block of AV transmission grade one to three), supraventricular and ventricular extrasystoles etc. Changes of the ECG tracing provide also some information which cannot be obtained by other means. PMID- 12744027 TI - [Evaluation of the ECG recording in abnormal positions of the heart in the thorax]. AB - Abnormal localization of the heart in the chest is a rare congenital developmental disorder (1,10). Even if the heart is in these instances normally developed, its abnormal position leads in case of its affection by disease to diagnostic and therapeutic difficulties, in particular when an invasive procedure must be used. With regard to the entirely different position of individual cardiac departments it is necessary to differentiate carefully between dextrocardia and dextroversion (15). Dextroposition is only a marginal problem. A very satisfactory diagnostic method in these anomalies is the ECG tracing, where we encounter quite typical pictures. Dextrocardia and dextroversion are characterized by a similar mirror image reversed tracing in the leads from the extremities. It differs, however, fundamentally in the chest lead tracings. In dextrocardia during tracing of standard thoracic leads we follow the predominant negative QRS complex in all leads; when including the dextrolateral thoracic leads the curve "becomes normal". In dextroversion, with regard to the anterior position of the left ventricle the dominating finding is a high R wave in all leads from the left and right side of the chest. PMID- 12744028 TI - [ECG in the diagnosis of supraventricular tachyarrhythmias]. AB - The development of catheter ablation techniques during the last decade provided new data about the mechanism of supraventricular tachyarrhythmias and at the same time, set new requirements for their classification. An accurate diagnosis of individual SVT can usually be made during an electrophysiologic study that precedes catheter ablation. Nevertheless, clinically acceptable differential diagnosis of SVT can be based on analysis of a standard 12-lead electrocardiogram. This may prove useful especially when selecting optimum antiarrhythmic drug according to a suspected mechanism of arrhythmia. At the same time, electrocardiogram during SVT serves as a recording of clinical arrhythmia for catheter ablation. At present, SVTs are divided into 3 main categories: 1. atrial tachyarrhythmias confined solely to atrial tissue, 2. tachycardias involving the AV junction, and 3. AV reentrant tachycardias involving one or more accessory connections with an electric impulse travelling between atria and ventricles. The first category can be further subdivided into: a) macroreentrant atrial tachycardias related to the presence of macroscopic anatomical or functional barriers; b) focal atrial tachycardias arising from a focus of abnormal automaticity or microreentry in the atrium; c) the syndrome of inappropriate sinus tachycardia resulting most probably from hypersensitivity to adrenergic stimulation; d) atrial fibrillation based on the existence of multiple wandering wavelets in the atria. Electrocardiographic differential diagnosis is predominantly based on an analysis of the standard 12-lead ECG. Principal diagnostic features include the presence and timing of the P waves in relation to the QRS complex. Additional criteria comprise the presence or absence of AV block during the tachycardia, an axis orientation of the P waves and their morphology, the appearance of QRS alternans or frequency of tachycardia. PMID- 12744029 TI - [Advantages and disadvantages of ECG diagnosis in left ventricular hypertrophy]. AB - Left ventricular hypertrophy (LVH) develops as a response to press or volume overload of the left ventricle. It is encountered in systemic hypertension, aortal stenosis and aortal insufficiency, mitral valve insufficiency, some congenital cardiac defects and hypertrophic obstructive cardiomyopathy. The most frequent cause of LVH in the adult population is hypertension. Prevalence of left ventricular hypertrophy in hypertensive individuals varies considerably, depending on the applied diagnostic method and characteristic of the group of hypertonic subjects. The presence of ECG signs of LVH is an infaust prognostic sign which at any level of blood pressure causes marked deterioration of the patient's prognosis. Classical ECG criteria for detection of LVH have a satisfactory specificity but low sensitivity. The sensitivity declines dramatically in the presence of obesity. The most sensitive but least specific criterium is high voltage in the precordial leads. The accuracy of ECG diagnosis of LVH improves considerably by multiplying the voltage by the duration of the QRS complex. Most probably the best results are obtained by Cornell's product. Falsely positive ECG signs of LVH (most frequently voltage criteria) are present in asthenic subjects. The author describes the case-history of a patient with hypertension who develops an ECG tracing imitating an infarction of the posterior wall and left ventricular hypertrophy with overburdening. PMID- 12744030 TI - [The electrocardiogram in patients with implanted cardiac pacemakers]. AB - In the submitted review the authors present electrocardiographic record of patients with atrial, ventricular and dual-chamber pacemakers. They also describe specially ECG findings in complications of cardiostimulation. Recommended procedures in these situation: changing pacemaker programme or electric cardioversion or surgical solution--reimplantation of pacemaker generator and/or lead or implantation of second lead. PMID- 12744031 TI - [The importance of electrocardiography in the development of new drugs--prolonged QT intervals as indicators of adverse effects]. AB - Both experimental animal studies and clinical observations have shown that drugs from various indication areas, such as psychotropic agents and antiarrhythmics can induce disturbances of cardiac rhythm and electrocardiographic abnormalities. Particularly, QT interval prolongation is associated with distinctive polymorphic ventricular tachycardias and often causes syncopes or cardiac arrest that represent a high risk of recurrent events including sudden death. In the last decade, several drugs in different indication areas, possessing these effects were withdrawn from the market and approvals were postponed or even refused. These events led to higher regulatory requirements on the cardiovascular safety of new drugs, with a special emphasis on prolongation of the QT interval. This article reviews the use of preclinical and clinical electrocardiography, as well as the current problems during development of novel drugs and future strategy in clinical studies. Even though ECG proved to be a powerful and reliable tool for detection of cardiac serious adverse effects during development of drugs, it also needs to be used with increased attention in all patients treated with new drugs and drug combinations in clinical and ambulatory practice in order to ensure their safe use in a wide population, as well as in patients with concurrent cardiac risk factors. PMID- 12744033 TI - [Will ECG combined with echocardiography detect viable myocardium in patients after myocardial infarct?]. AB - The objective of the submitted trial was to use combined examination of ECG at rest and after exercise to detect a viable heart muscle in patients with a postinfarction Q wave and ST elevations. The authors found that according to the character of changes of the ST elevation in ECG after exercise it is not possible to detect patients with a viable myocardium. During the combined procedure using analysis of the kinetic disorder at rest and increase of the ST segment in ECG after exercise it is possible in patients with hypokinesia or akinesia at the site of the Q wave to differentiate successfully 66% patients with a viable heart muscle. This combined procedure can serve as a selective test for implementation of further more accurate diagnostic tests for the detection of a viable heart muscle because in common clinical practice the above data are available in these patients before indication of more complicated examinations. PMID- 12744032 TI - [Relation between anthropometric indicators and electrocardiogram variability]. AB - AIM: To investigate which combination of anthropometrical parameters influence significantly the variability of the magnitude spatial vectors of atrial (sPmax), activation and ventricular (sQRSmax) activation and ventricular depolarization (sTmax). Quantification of these relationships can improve the reliability of criteria for normal values of ECG and VCG characteristics. SUBJECTS AND METHODS: The study group consisted of 195 healthy boys and men without signs of cardiac disease, with normal ECG and VCG, aged from 7 to 70 years, (median = 19 years). Electrocardiologic variables were obtained by Frank lead system in seated persons from a single heartbeat by the computer system Cardiac 128--PC (METE, Prague) in mid respiration. 30 anthropometrical measurements were taken including biceps, triceps, subscapulare and suprailiacare skinfold thicknesses. Proportionality of body and thoracic shape was evaluated according indexes computed from body height, weight (Body mass index, Rohrer and Ponderal index), circumferences (Relative abdominal circumference, Pignet index), anteroposterior and transversal dimensions (Thoracic index, relative sagittal diameter). Values of the 15th and 85th percentil of relevant indexes were taken as criteria for stratification of the sample into three subgroups demarcating the constitutional types. Differences in maximal spatial vector magnitudes between these subgroups were tested by variance analysis ANOVA. Relationships to anthropometrical and vectorcardiographic parameters were expressed also by regression equations. RESULTS: The average magnitudes of maximal spatial vectors increased during maturation and early adulthood: sPmax, approximately up to 17 years, sQRSmax and sTmax up to 22 years. The magnitude of ventricular vectors decreased after this period. With increasing endomorphy, which is indicated by values of Rohrer index, relative abdominal circumference and fat percentage, magnitudes of sPmax, s QRSmax and sTmax decreased (p < 0.001) on the other hand they increased with higher linearity (Ponderal index) and index of fitness (Pignet index). The greatest differences (by 50%) were found in values of sPmax, whereas distributions of ventricular vectors sQRSmax differed by 20-30%. CONCLUSIONS: The variability of magnitudes of maximal spatial vectors was significantly influenced by the proportionality of body constitution independently of age. The tightest relationship of sTmax was found with fat percentage. Influence of anthropometrical parameters was most pronounced in younger adulthood and should be taken into account when evaluating of electrocardiograms and vectorcardiograms. PMID- 12744034 TI - [Evaluation of the ECG in epidemiologic studies of cardiovascular diseases--the Minnesota code]. AB - The authors evaluate the importance of the Minnesota code for description of the ECG curve in epidemiological surveys. The ECG tracing is not evaluated clinically but is only described and changes are summarized into categories marked arbitrarily as minor and major ischaemic changes. In these categories their predictive value is assessed on a longitudinal basis. It was revealed that minor and major ischaemic changes have a predictive value for cardiovascular morbidity and mortality similarly as other risk factors of ischaemic heart disease. The authors mention some epidemiological surveys where the Minnesota code was used. Assessment of the ECG tracing and its evaluation by the Minnesota code should be done by a person trained in this method. At present it is possible to use also computer evaluation of ECG tracings by the Minnesota code. PMID- 12744035 TI - [The importance of electrocardiographic changes at rest during exercise in deciding to perform immediate ("ad hoc") percutaneous transluminal coronary angioplasty]. AB - In 100 patients (75 men and 25 women, mean age 53 years) who had "ad hoc" percutaneous transluminal coronary angioplasty (PTCA) on one or two vessels retrospectively the ECG records were evaluated (signs of necrosis or ischaemia) at rest and/or after exercise and compared with the finding of major obstruction (complete occlusion, stenosis > 50%) on coronarography. In obstruction of the ramus interventricularis anterior the positive ECG finding was consistent with the area of the impaired blood supply in 87%, in case of obstruction of the right coronary artery in 79%, in obstruction of the ramus circumflexus in 90%. In concurrent obstruction of two coronaries despite a 100% positive ECG no close topical correlation was found between coronarography and ECG. On the whole no reliable consistency was found between ECG and kinetic disorders detected by echocardiography. The results indicate that topical analysis of ECG ischaemic findings in patients with coronary arterial disease before planned coronarography contributes to the decision to make "ad hoc" PTCA in affections of one artery. In affections of two arteries ECG is of no particular value. PMID- 12744036 TI - [Electrocardiography in the diagnosis of genetic heart diseases--the Brugada syndrome]. AB - Electrocardiography celebrates its 100th anniversary but is still a useful and widely used examination method in cardiac diseases. Its importance can be demonstrated on its part in the diagnosis of a relatively newly described disease presented in this paper. It is a genetically conditioned anomaly of sodium channels of the cell membrane, called Brugada syndrome. The latter has a typical electrocardiographic tracing--elevations of the ST segment in leads V1-V3 along with a finding reminding of a right bundle branch block, absence of structural cardiac disease and an increased incidence of serious ventricular arrhythmias and sudden arrhythmic death. PMID- 12744037 TI - [ST segment elevation in the ECG stress test in coronary disease]. AB - The authors investigated in 1986-1991 a group of 122 men after myocardial infarction, their mean age was 56 years, the follow up period 35 and at least 24 months. During the ergometric test "symptom limited" 18 days after the attack it was negative in 55% of the men, in 28% positive (depression of ST and/or coronary pain (and in 17% there were elevations of ST above the pathological Q wave. By the end of the follow up period nobody with a negative baseline test died. 6% with a positive test died, 62% patients with elevation of the ST. All patients with ST had elevations an impaired regional ventricular motility with a low ejection fraction (32%) during radionuclide ventriculography. At the time of examination the availability of coronarography and possible revascularization was minimal. Therefore even a retrospective group is valuable. Patients with elevation of ST after exercise have a generally poorer prognosis and are therefore indicated as a priority group for coronarography or possibly revascularization. ST elevation after a load without a pathological Q wave is less frequent and need not be associated with impaired regional motility and may be the manifestation of a coronary spasm or close proximal coronary stenosis, and is also indicated for angiographic examination, and deepening on the result, for revascularization. PMID- 12744038 TI - [Heart rate turbulence--a new ECG predictor for risk of sudden death]. AB - Heart Rate Turbulence (HRT) is a newly described physiological chronotropic response of sinus rhythm following a single ventricular premature beat (VPB) consisting of early acceleration and later deceleration of heart rate. Using two large independent cohorts of postinfarction patients, the absence of HRT was retrospectively validated to be a potent multivariate risk predictor, stronger than a number of currently available risk stratifiers. Although exact pathophysiological mechanism of HRT remains speculative, it is now believed that HRT arises from the haemodynamic changes and baroreceptor reflexes that occur following a VPB. Therefore, HRT descriptors (Turbulence Onset and Turbulence Slope) may serve as very reasonable, Holter--based surrogates of baroreflex sensitivity available in clinical practice. The aim of this review is to summarise the current knowledge on pathophysiological mechanisms of HRT and to discuss practical problem of its detection. PMID- 12744039 TI - [ECG mapping in clinical practice]. AB - First the authors present a review of important cornerstones in the history of the electrocardiogram (ECG) and ECG mapping. The first to describe the electric cardiac field based on twenty ECGs was A.D. Waller in 1889. The decisive cornerstone for practical use was the introduction of a string galvanometer in 1901 by W. Einthoven and his triaxial lead system. Another very important cornerstone in the development of ECG were the findings of F.N. Wilson. Merits as regards the development and application of ECG mapping are due to B. Taccardi. Workers of the Second Medical Clinic in Prague enhanced after 15 years of studies and comparison of ECG maps with coronarographic findings in subjects with ischaemic heart disease (IHD) and microvascular coronary dysfunction (syndrome X- SyX) substantially the specificity of this method in impaired myocardial vascularization. Better diagnosis was achieved by introduction of diagnostic tests which influence coronary vascularization such as e.g. hyperventilation, as well as other tests. After their application progression of chronic myocardial ischaemia occurs, e.g. by the mechanism of the "steal phenomenon" or restriction of the microcirculation after hyperventilation in patients with SyX. Furthermore the authors present examples of ECG maps after PTCA, after application of diagnostic tests in IHD and SyX and also regression of myocardial ischaemia after marked reduction of total cholesterol. PMID- 12744040 TI - [Effect of adrenergic stimuli on electrocardiographic and vectorcardiographic characteristics of ventricular repolarization]. AB - Autonomic effect of various stimuli on haemodynamic variables is usually tested by changes in blood pressure (BP) and/or heart rate (HR). It is known that increased sympathetic drive of ventricles can interfere with repolarization process. This study was focused on reactive changes of maximal spatial T vector (sTmax), R-R and QTc intervals, in relation to BP changes in 79 boys and men, averaged age 17 +/- 2 years, 36 from them were adolescents with elevated BP (high normal or hypertension I according to WHO/ISH 1999) (ZTK), 19 normotensives (NTK), and 24 normotensive sportsmen. R-R, QTc intervals and maximal spatial T vector were recorded by a PC (Cardiag METE, Prague) with Frank lead system while sitting in mid-respiratory position, during mental arithmetic (MA), after handgrip and during passive head-up tilting to 60 degrees. BP was measured simultaneously by a cuff sphygmomanometer, using phases 1 and 5 of Korotkoff sounds. MA resulted in significant BP increase in all subgroups, however the reactive changes of systolic BP as well as magnitude of R-R shortening, sTmax decline were about two times higher, and in the case of QTc lengthening three times higher in sportsmen. Handgrip provoked in all subgroups in average the less reactive changes of diastolic BP, sTmax, R-R, and QTc intervals. Head-up tilting evoked in all subgroups a significant increase of diastolic BP that was again higher in sportsmen vs. ZTK and NTK. R-R intervals became significantly (p < or = 0.02) and more often shortened in sportsmen vs. ZTK and NTK. Relatively more evident decline of sTmax (more than 20%) in ZTK was the highest change from all observed parameters to all tested stimuli. Our results indicate that for analysis of effect stimuli, which modulate balance in autonomic nervous system, it is helpful to pay attention also to the parameters of repolarization process that may represent a sensitive indicator of sympathetic tonization in myocardial ventricles. PMID- 12744041 TI - [The intracardiac electrocardiogram in the diagnosis of arrhythmias]. AB - During the last 15 years in arrhythmology brisk development of the use of endocardial electrocardiograms (IECG) occurred. At first the objective was to describe the characteristics of the conduction system of the heart (recording of Hisogram), next assessment of pathological features of the conduction system (such as bradyarrhythmias), electric activation of the myocardium in existing arrhythmias with the outcome of the diagnosis and accurate localization of additional pathways in endocardial mapping. The next logical step reacting to knowledge of intracardial mapping was the development of new therapeutic methods- DC and subsequently radiofrequency ablation--non-pharmacological therapeutic antiarrhythmic approaches which are by now commonly used in electrophysiological catheterization laboratories of cardiological departments. Some contemporary pacemakers can also record intracardial atrial and ventricular IECG and make it possible to use additional antiarrhythmic stimulation. A dominant position is held by the IECG in implantable cardioverters-defibrillators where on the high standard recording of IECG a considerable proportion of detection and decision making algorithms of this equipment is based. Recording IECG is an integral part of clinical arrhythmological practice from the diagnostic as well as therapeutic aspect. PMID- 12744042 TI - [Three-dimensional presentation of the electrocardiogram: new possibilities in comparative studies using cardiologic imaging methods]. AB - The aim of the study was to demonstrate a biomathematical model for the topographic presentation of orthogonal electrocardiogram DECARTO, to present the possibilities for visualisation and processing of ECG in relation to others cardiological imaging methods. In DECARTO (Dipolar ElectroCARdioTOpography), the orthogonal ECG is used as an input signal, the signal is processed by the means of a biomathematical model and presented as areas of activated points on a spherical surface closely surrounding the heart, so-called decartograms. Decartograms can be visualised using the projection of the spherical image surface onto geometrically defined planar or 3D surfaces. The selection of the surface depends on the purpose of ECG imaging, on the evaluation of advantages and limitation of the particular possibilities, such as the degree of deformation, and their comparability with other imaging methods. Another possibility of 3D visualisation of decartograms is the utilisation of time as a 3rd dimension of the space. Additionally to analytically defined geometrical surfaces, it is possible to project decartograms onto a real surface of the heart, constructed from data of other image techniques. In this paper we present a superposition of decartogram and SPECT imaging, which combines graphic presentation of two diagnostic methods providing different but complementary information on the heart. The combination of comparable information allows to study the relationship between electrogenesis and perfusion of myocardium and its metabolism, to study diagnostic and prognostic importance of both agreements and disagreements of used methods. The fusion of information on the structure and function of the heart could potentially increase the diagnostic power of both methods. PMID- 12744043 TI - [Lead systems in orthogonal electrocardiography]. AB - The paper deals with evaluating quality of some orthogonal and pseudoorthogonal electrocardiographic lead systems as Frank lead system, signals derived from standard electrocardiographic leads by Levkov's transform and some signals of the standard 12 lead system that roughly fulfill condition of geometrical orthogonality. The signals of the lead systems were assessed by means of a mathematical criterion of two vector orthogonality which was originally proposed for purposes of electrocardiography and proved by Netusil. Experiments done with signals of the CSE database of short-time rest ECG records have revealed that the best ECG lead orthogonality has been found for signal sets of I-aVF-V2, and I-III V2 leads, resp. Unfortunately nearly the worst results were obtained for the widely used Frank orthogonal lead system. It means that this lead system can lead (under some specific conditions) to missing diagnostically important information. PMID- 12744044 TI - [Orthogonal electrocardiography]. AB - The orthogonal electrocardiography was a further development of Einthovens conception of electrocardiography. It eliminates the redundancy of the 12-lead ECG and offers a more precise and illustrative image of the model of an equivalent dipole. It was the prerequisite of the development of vectorcardiography. It improves the diagnostic performance of electrocardiology, especially by decreasing the spread of parameters used to characterize normal cardiac electrogenesis. It may be assumed that clinical electrocardiography will not petrify its standard 12-lead system but will utilize the possibilities offered by the more rational lead systems of orthogonal electrocardiography, as well as its enrichment by vectorcardiographic presentation of the cardiac electric field. PMID- 12744045 TI - [The ECG and acute coronary syndromes in patients under 40 years of age--case reports of an obese smoker in comparison with an active female athlete]. AB - The authors describe two cases of acute coronary syndrome by patients to the age of 40, of which anamnestical data and clinical manifestation were quite different. According to present recommendation of European Society of Cardiology neither one of them fulfilled standards for acute or created myocardial infarction. The discovery of ECG changes or echocardiographical findings lead to an introducing of coronarography examination that proved the important stenosis of coronary arteries and both cases were closed as a created (case 1) and acute (case 2) myocardial infarction. PMID- 12744046 TI - [The electrocardiogram in primary pulmonary hypertension]. AB - Primary pulmonary hypertension is a rare disease characterized by a rise of arterial pressure in the pulmonary artery without a known cause. The authors define, based on a case-history, the etiopathogenesis of the disease, its diagnosis and approach to treatment. They analyze the contribution of ECG examination for detection of right ventricular hypertrophy and for monitoring the therapeutic success with the finding above all of repolarization changes in the right precordium. The patient had a transplantation of the lungs. PMID- 12744047 TI - [Complete atrioventricular block (with Adams-Stokes syndrome) in systemic connective tissue disease. Electrocardiographic development in three patients]. AB - The association of high grade atrioventricular heart block with systemic connective tissue diseases is very rare. To date, only sporadic case reports or reviews appeared in the literature. Three cases of such association observed by the author are described here. The patients were a 51 years old man with systemic sclerosis and two women, a 64 years old patient with visceral nodous seropositive rheumatoid arthritis, and second was a 74 years old patient with systemic lupus erythematosus, the oldest of the group of patients with this disease. In two of the three patients, Adams-Stokes attack was a cause of death. The author did not observe this high grade atrioventricular block in patients with systemic connective tissue diseases the last 32 years. First and second grade heart blocks were, however, o chi asionally seen and responded well to the treatment of the systemic disease. Thus, in patients with systemic connective tissue diseases and emergency symptoms, it is recommended to consider also this rare association. While, two decades ago, patients frequently died after the Adams-Stokes attack, the treatment of the high grade block is now successful due the permanent cardiostimulator. In the last years, there are almost no reports about the association of systemic connective tissue diseases with high grade heart block, presumably because of the efficient new treatment approached to systemic diseases including modern immunomodulation drugs. PMID- 12744048 TI - [The deceptiveness of the electrocardiographic image in patients with Wolff Parkinson-White syndrome]. AB - The urgent character of the clinical importance of the Wolff-Parkinson-White syndrome is among others also due to some treacherous characteristics of the electrocardiographic tracing. Three of them are mentioned in the submitted paper. The lability of the electrocardiographic tracing--spontaneous and induced--makes sometimes early detection difficult. The risk of confounding the ECG tracing of Wolff-Parkinson-White syndrome with myocardial infarction is considerable and sometimes underrated. Diagnosis of the ECG picture of myocardial infarction in a subject with Wolff-Parkinson-White syndrome may be associated with great difficulties. The electrocardiographic picture of the Wolff-Parkinson-White syndrome must be evaluated always with knowledge of the general clinical manifestations to prevent diagnostic, therapeutic and assessment errors. PMID- 12744049 TI - [Is the negative T-wave on the ECG always a sign of ischemia? (human stress cardiomyopathy?) ]. AB - A 74-year-old woman was admitted on account of chest pain which developed after a death in the family. On the ECG on admission there were negative T waves (the picture of extensive subapicardial ischaemia). Coronarography made on the subsequent day was quite normal. The laboratory finding (CK, CK-MB, AST) did not suggest an acute coronary attack. Ultrasound examination of the heart revealed only slight hypokinesia of the anterior wall. ECG returned to normal within 3 months. Four years later the patient is asymptomatic. The ECG finding and clinical course suggest the clinical entity of "stress cardiomyopathy", which is not a well known and unequivocally accepted diagnosis. PMID- 12744050 TI - [The signal-averaged ECG in long-term monitoring of patients with fulminant myocarditis]. AB - A case of a 24 year-old patient with fulminant myocarditis is described. The lymphocytic myocarditis was proved by endocardial biopsy. The patient had been followed for 15 months clinically, by transthoracic ultrasound and by the signal averaged ECG. The clinical and ultrasound parameters of the left ventricular systolic dysfunction subsided within 6 months, STT changes on the routine 12-lead ECG within 2 months. On the signal-averaged ECG, changes were observed in both the time and in the frequency domain. When the results of the initial examination (6 weeks after the beginning of the disease) were compared with the examination at the 7th month of the disease there was a prolongation of the QRS duration (from 95 ms to 104 ms), prolongation of the duration of the filtered ECG below 40 microV (from 21.5 to 36.2 microV) and the decrease of the total sum of the potential in the terminal part of the QRS. Most apparent was the decrease in the total sum of the high-frequency components (filter 70-250 Hz) of the QRS (RMSt: from 53.3 microV to 37.9 microV at the same noise level). In contrast to the changes in the time domain parameters there was no tendency in improvement of the RMSt. In patients with myocarditis the signal averaged ECG may prove to be a suitable tool for the long-term follow-up of the subclinical structural abnormalities of the left ventricular wall. PMID- 12744051 TI - [Signs of extreme vagotonia in the electrocardiogram of an Olympic 5km running champion]. AB - Hypokinesia (lack of exercise)--a disease caused by lack of physical activity leads among others to a predominance of the sympathetic nerve in the circulation. Its manifestations are briefly described according to Kraus-Raab's hypothesis of 1961. The opposite of a hypokinetic man are trained (hyperkinetic) sportsmen, in particular long-distance runners. Their neurovegetative balance is shifted to the side of the vagus (parasympathetic nerve). This is described as training vagotonia. On the example of ECG tracings of an Olympic 5 km runner, the Tunisian Mohamed Gammoudi, the author describes its manifestations at rest, during the orthostatic test and during work, i.e. the step test. These manifestations include in particular bradycardia of 32 beats per minute, prolonged atrioventricular conduction > 0.2 sec., a long period of the electric systole = 0.44 sec. and an enormously prolonged electric diastole T-P = 1.1 sec. This long diastolic filling stage makes it possible in particular during physical work to increase the systolic and diastolic output to double the values of a healthy non trained heart. Training vagotonia is usually not the source for manifestation of dangerous arrhythmias, as it may be the case in constitutional vagotonia. So far expert comparison of these two types of change to parasympathic tonus is lacking, similarly as a comparative study with medicamentously influenced vagotonia by means of beta-blockers. PMID- 12744052 TI - [Professor Valentina Il'inichna Puzik's life and creative work]. PMID- 12744056 TI - [Current views of tuberculosis inflammation]. AB - The value of histological and histochemical studies in the diagnosis of a phase of tuberculosis progression or healing is shown. Electron microscopic study of tuberculous inflammation in different phases of its evolution evaluated the functional status of cellular elements of the lung and granuloma. The body's antituberculous resistance due to molecular genetic mechanisms is realized through intercellular interactions and macrophageal functions. Immune macrophages are characterized by a higher metabolic activity, they suppress the intracellular multiplication of Mycobacterium tuberculosis (MBT) and are more protected from their toxic action. The pathogenetic mechanisms responsible for caseous pneumonia were studied. Three stages of evolution of the process: Stage 1 is the breakdown of defense and adaptive mechanisms: disorganization of connective tissue and alveolar parenchyma; enhanced permeability of blood and lymphatic microvascular walls with developed interstitial and intraalveolar edema, plasma and fibrin exudation, fibrinoid swelling of collagenous fibers, and their lysis; occurrence of lung parenchymal microinfarcts and infarction-pneumonia; type 2 alveolocytic dysfunction with surfactant destruction; Stage 2 is the breakdown of local immunity; exudative and alterative tuberculous inflammation with involvement of immunocompetent organs; suppressed T-cellular immunity, a shift of a T helper/T suppressor ratio to the latter, lymphopenia; impaired intercellular interactions, cellular apoptosis in blood and inflammation areas, and suppressed granulomatous reaction; inhibited L transformation of Mycobacteria tuberculosis, intensive MBT multiplication in the foci of tuberculous inflammation, particularly those which are resistant to many antibiotic drugs, a larger number of associations of the nonspecific microflora and fungi. Stage 3 is caseous pneumonia and generalization of a tuberculous process: a predominance of an alterative reaction of inflammation; the presence of allergic and caseous and necrotic vasculitis, bronchiolitis, and endo-panbronchitis; depressed granulomatous reaction; the development of acute alterative sequestrating pneumoniogenic caverns. Histological, histochemical, and electron microscopic studies of tuberculous inflammation may specify the mechanisms of the pathogenesis of tuberculosis and may serve as the basis for early diagnosis of the disease and for timely correction of performed treatment in order to enhance its efficiency. PMID- 12744057 TI - [Immunopathogenetic mechanisms of action of BCG and BCG-M vaccines]. PMID- 12744058 TI - [Tuberculosis in adolescents: diagnosis, clinical picture, and treatment]. AB - The paper is dedicated to the anniversary of Professor V. I. Puzik, Honored Science Worker. It gives the data of morphological studies from her dissertation and some sections of the dissertations supervised by V. I. Puzik. The paper also presents data on the diagnosis, clinical picture, and treatment of tuberculosis in adolescents. The problems of early detection, differential diagnosis, the significance of concomitant diseases and social factors in the course of tuberculosis are discussed. Emphasis is placed on the importance of drug resistance in adolescent patients with tuberculosis, which has been revealed in 63.3% of cases. Complex treatment with essential, reserve, and combined antituberculous drugs is recommended in relation to the sensitivity of Mycobacterium tuberculosis. It is also advisable to treat concomitant diseases and to prescribe pathogenetic agents enhancing the body's overall resistance. PMID- 12744059 TI - [Role of ultrasmall forms of Mycobacteria in the pathomorphology of tuberculosis]. AB - Microbiological and histological studies made in patients with active tuberculosis revealed that 65.5% of cases had revertants from ultraminor forms (UMF) of Mycobacteria tuberculosis (MBT), which are typically bacterial in shape and referred to as representatives of the genus Mycobacterium tuberculosis. Cultivation of the diagnostic material containing MBT UMF the enriched liquid nutrient medium could detect rod-shaped bacterial forms of MBT in 18% of the patients with tuberculosis. Inoculation of the MBT UMF-containing material isolated from patients with tuberculosis to guinea-pigs revealed productive sarcoid-like changes in the organs of the experimental animals in 30.6% of cases. A postmortem study of the organs of experimental animals inoculated the material containing "visible, but not growing" mycobacteria detected paraspecific tissue reactions and infiltration of lung tissue by multiple cells of the macrophageal series. PMID- 12744060 TI - [Significance of molecular genetic and immunological techniques in risk groups and in children with tuberculosis]. AB - A total of 188 children and adolescents were examined. In all the children, blood Mycobacterium tuberculosis (MBT) DNA was determined by polymerase chain reaction (PCR) and MBT antigens (AG) and antibodies (AB) were by enzyme immunoassay. The studies have shown that it is expedient to concurrently determine MBT DNA and MBT AT in order to identify local forms of tuberculosis in children from risk groups. If the tests are positive, a comprehensive examination for tuberculosis is required; the presence of the syndrome of common disturbances is generally associated with tuberculous infection. When a local form of tuberculosis is excluded, preventive chemotherapy should be performed. Further negative tests for MBT DNA and lower MBT AT may be a criterion for the efficiency of preventive treatment. In children with tuberculosis, the results of repeated blood and urine tests for MBT DNA provide a way of evaluating the course of a tuberculous process and the efficiency of chemotherapy. PCR used to determine blood and urine MBT DNA is a highly specific test as positive results were in 79% of the children with tuberculosis. PMID- 12744062 TI - [The morphological differences in tuberculous inflammation foci, which reflect the body's immunological responsiveness]. AB - A comparative study of morphological reaction in the foci of tuberculous inflammation in decreased patients with caseous pneumonia (n = 2), fibrocavernous pulmonary tuberculosis (n = 19), disseminated tuberculosis (n = 5) and on the material of resected parts of the lung (infiltrate in a phase of separation (n = 2), tuberculomas (n = 22), caverns (n = 5)) has shown that exudative and necrotic reactions with minimum discriminating signs and no epitheliocytes, by accompanying by atrophy of the paracortical area of intrathoracic lymph nodes reflect immediate hypersensitivity. The decrease in the rate of exudative necrotic reactions, the increase in fibroplastic processes, and the appearance of epitheliocytes suggest that immediate hypersensitivity is added by delayed hypersensitivity. Pronounced fibroplastic reactions around the foci of caseosis and on the walls of tuberculous caverns, involvement of epitheliocytes in cellular reactions point to the fact that a tuberculous process develops in delayed hypersensitivity. PMID- 12744061 TI - [Acutely progressive pulmonary tuberculosis: morphological and bacteriological features]. AB - The data of autopsies (n = 431) and morphological (n = 52) and bacteriological (n = 47) studies of the operating material obtained from the lungs of patients with tuberculosis during 1980-1989 and 1990-1999 were analyzed at the Research Institute of Pulmonology and Phthisiology, Republic of Belarus. The analysis revealed an increase in the incidence of acutely progressive forms of pulmonary tuberculosis. Autoptic data indicated that in 83.7% of cases, acute progressive tuberculosis was associated with the development of caseous pneumonia as an independent tuberculous process (31.7%) and as a phase of ensuing development of other forms (52.0%). The distinctive morphological features of today's caseous pneumonia were as follows: a great extent of lesion, an unrestrained progression of a process, a predominance of an alterative-and-exudative component of tuberculous inflammation, a rapid decay and suppuration of caseous masses, no differentiation of caseous and pneumonic foci, and suppressed cellular reactions of tissue immunity. The morphological and bacteriological studies of the operating lung material from patients with pulmonary tuberculosis suggested that there was a predominance of a progressive disseminating process at the moment of surgery. PMID- 12744063 TI - [Results of studying of the pathomorphism of pulmonary tuberculosis in Iakutiia]. AB - The paper summarizes the results of studies into the pathological anatomy and pathomorphism of pulmonary tuberculosis in Yakutia, which have been conducted by the Department of Anatomy, Yakutsk State University Medical Institute, over 36 years. A morphological comparative analysis of autopsies of individuals who died from different forms in the pre-antibacterial and antibacterial periods and surgical materials were used as a major source of data. More than 1,000 autopsies made it possible to define a wide range of behavioral patterns and the morphological features of pulmonary tuberculosis under the extreme climatic conditions of the Far North. These findings have made a significant contribution to the development of the new scientific school Northern Phthisiology. PMID- 12744064 TI - [Clinical manifestations and specific features of the course of adenogenic tuberculosis in adults]. AB - The paper analyzes the clinical manifestations of adenogenic tuberculosis, the methods of its diagnosis and treatment in 44 patients (15 males and 29 females) aged 18 to 75 years. The signs of an active tuberculous process were identified in the bronchus and lung of 33 patients and the remaining patients had posttuberculous adaptogenic changes. A bronchological study revealed infiltrative bronchial tuberculosis in 15 patients, bronchial fistula in 4, inflammatory and cicatrical stenoses in 22, broncholiths in 2, and nonspecific bronchitis in 5. The patients with specific active changes received antituberculous chemotherapy with 4 drugs; those with nonspecific lesions had antirecurrent chemotherapy and, in case of an exacerbation of nonspecific recurrent pneumonias and chronic bronchitis, used antibiotics mainly as inhalants. PMID- 12744065 TI - [Morphological study of tuberculin reaction to determine the body responsiveness in tuberculosis]. PMID- 12744066 TI - [To the problem of abacillary caverns in patients with pulmonary tuberculosis]. AB - Less interest has been recently shown in the so-called openly negative syndrome in patients with pulmonary tuberculosis, which occasionally results in healing of tuberculous caverns to form a cystoid cavity. The materials presented in this paper suggest that in the 1960s the openly negative syndrome occurred after chemotherapy in 8.7% of the patients registered at a tuberculosis facility for destructive pulmonary tuberculosis. This syndrome persists for 3 years or more (according to the data of bacterioscopy, sputum inoculation and biological test on guinea pigs). Of great interest are the data of histological study made under the head of Prof. V. I. Puzik on experimental animals. The absence of specific reactions in response to a biological test and the presence of significant lymphoid focal and diffuse infiltration in the lungs, hyperplasia of follicles of the spleen and lymph nodes allow them to be regarded as immunomorphological reactions in response to the protein components of destroyed Mycobacterium tuberculosis (MBT). The long existing openly negative syndrome is also presented observed in patients with pulmonary tuberculosis but only in those who isolate MBT sensitive to antituberculous agents. In this connection, it is not inconceivable that special reparative processes that do not result in the long (3 years or more) existing openly negative syndrome in patients with destructive pulmonary tuberculosis who isolate resistant MBT forms. PMID- 12744067 TI - [The morphofunctional state of the surfactant system in pulmonary tuberculosis]. PMID- 12744068 TI - [Segmental pathology of pulmonary tuberculosis: resectional and sectional data]. PMID- 12744069 TI - [Data on the comparison of radionuclide and morphological studies in pulmonary tuberculosis]. PMID- 12744070 TI - [Cytomorphologic changes detected in bronchial biopsy specimens in patients with alveolitis]. AB - Eighty-nine patients with alveolitis were followed up. Of them, 60 patients had exogenous allergic alveolitis (EAA) and 29 had idiopathic fibrosing alveolitis (IFA). Transbronchial lung biopsy (TLB) proved to be the most informative method for alveolitis. At the same time a cytomorphologic study of TLB specimens revealed exogenous allergic alveolitis in all phases of the disease in 91.2% of the cases. In idiopathic fibrosing alveolitis, the informative value of TLB was 59.7%. PMID- 12744071 TI - [Some pathomorphological features of pneumofibrosis in tuberculosis]. AB - The processes of organization and fibrosing were investigated in the tuberculosis involved and non tuberculosis-involved lung tissue of the lungs and their parts resected in 307 patients with different forms of tuberculosis by histological, histochemical, luminescence microscopic, biochemical, and morphometric studies. When tuberculous lesions develop and begin fibrosing, there was impairment of final formation of collagenous fibers in their inner area due the low proliferative capacity of epithelioid and giant cells, shortage of fibrogenic cells and precipitation on the argyrophilic fibrillary network of tissue degradation products. Etiologically, pathogenetically, and histogenetically, the development of tuberculous lesions gives rise to posttuberculous fibrosis while that of extratuberculosis lesions leads to metatuberculous fibrosis due to the modification and coarsening of preformed connective tissue. The formation of fibrosis is caused by a combination of definite factors: clinicomorphological forms of tuberculosis, long-term history of the disease, the extent of lung tissue decay, old age, the pattern of antituberculous treatment (regular or regular), the extent of the impaired capillary network, the type of arbitrarily fibrogenic cells functioning inside and outside the area of inflammation. PMID- 12744072 TI - Molecular features of the cytolytic pore-forming bacterial protein toxins. AB - The repertoire of the cytolytic pore-forming protein toxins (PFT) comprises 81 identified members. The essential feature of these cytolysins is their capacity to provoke the formation of hydrophilic pores in the cytoplasmic membranes of target eukaryotic cells. This process results from the binding of the proteins on the cell surface, followed by their oligomerization which leads to the insertion of the oligomers into the membrane and formation of protein-lined channels. It impairs the osmotic balance of the cell and causes cytolysis. In this review the molecular aspects of a number of important PFT and their respective encoding structural genes will be briefly described. PMID- 12744073 TI - Hybrid antibiotics. AB - Hybrid antibiotics that do not occur in nature have been obtained by combining structural genes of antibiotic producers. Some of these substances were effective against pathogenic microorganisms resistant against antibiotics produced by the parent strains. The majority of hybrid antibiotics were obtained by combining genes encoding polyketide synthases. Hybrid peptides with new biological properties have also been synthesized. PMID- 12744074 TI - Biodegradable plastics from renewable sources. AB - Plastic waste disposal is a huge ecotechnological problem and one of the approaches to solving this problem is the development of biodegradable plastics. This review summarizes data on their use, biodegradability, commercial reliability and production from renewable resources. Some commercially successful biodegradable plastics are based on chemical synthesis (i.e. polyglycolic acid, polylactic acid, polycaprolactone, and polyvinyl alcohol). Others are products of microbial fermentations (i.e. polyesters and neutral polysaccharides) or are prepared from chemically modified natural products (e.g., starch, cellulose, chitin or soy protein). PMID- 12744075 TI - Antimycobacterial activity of basic ethyl esters of alkoxy-substituted phenylcarbamic acids. AB - A series of 124 basic ethyl esters of alkoxy-substituted phenylcarbamic acids with the alkoxy group in position 2, 3 or 4 on the phenyl ring, and basic substituents attached to the ethyl moiety in position 2, were evaluated for in vitro antimycobacterial activity against strains of Mycobacterium tuberculosis, Mycobacterium kansasii and Mycobacterium avium. In vitro antimycobacterial activity becomes higher with increasing hydrophobic properties of the alkoxy groups. The p- and m-substituted derivatives were more active than the o substituted ones. Direct relationship between the structure of the basic substituents and the activity was not found. PMID- 12744076 TI - Simple synthesis of novel diphenylsulfapyrimidine acetates from chalcones and their antimicrobial activity. AB - Given the significant low yield (19-43%) in reported results on the cyclocondensation of sulfaguanidine acetate with chalcones, a careful reinvestigation was carried out. A new series of chalcones, bearing electron attracting groups in the aromatic moiety, have been used as precursors in the synthesis of diphenylsulfapyrimidine acetates with good yield. All synthesized compounds were active against G(+)- and G(-)-bacteria, and fungi. Combination of substituents (Cl, OMe, NO2, etc.) enhanced antimicrobial activity. Derivative with two NO2 groups exhibits an activity comparable with sulfadiazine. PMID- 12744077 TI - Evidence for chromosomal determination of fungicidal activity in strains of Lactobacillus brevis and Lactobacillus fermentum isolated from fermented foods. AB - The genetic basis of the fungicidal activity of strains of Lactobacillus brevis and L. fermentum isolated from indigenous fermented foods was determined. A 5.5 kb plasmid was isolated from L. brevis while L. Fermentum was found to harbor no plasmid. Plasmid curing indicated no correlation between the plasmid and the fungicidal activity of the Lactobacillus species. The fungicidal activity of the isolated organisms can be supposed to be mediated by the chromosome. No antibiotic resistance genetic markers were detected on the plasmid and hence it was classified as cryptic. PMID- 12744078 TI - Evaluation of Tolypothrix germplasm for phycobiliprotein content. AB - Twenty Tolypothrix strains, including 15 strains of T. tenuis, three strains of T. ceylonica and one strain each of T. nodosa and T. bouteillei, were evaluated for their phycobiliprotein content and composition. Significant differences among the Tolypothrix strains were found at both inter- and intra-specific levels in the production of phycobiliprotein constituents--phycocyanin (PC), allophycocyanin (APC) and phycoerythrin (PE). Four specific parameters, viz. PC or PE content, total phycobiliprotein and total protein content, and percentage of phycobiliproteins, in a mixture of total proteins were used to select four T. tenuis and one T. ceylonica strain as useful for phycobiliproteins production. PMID- 12744079 TI - Ultrastructure of the cyanobacterium Nostoc muscorum and exploitation of the culture for hydrogen production. AB - An effect of various physico-chemical parameters on nitrogenase-catalyzed oxygen free hydrogen production by Nostoc muscorum was demonstrated. More hydrogen was produced in the light than in the dark. Optimum temperature was 40 degrees C Various sugars increased hydrogen production whereas on easily metabolized nitrogen sources it was inhibited. The production was sensitive to salinity and Fe3+, Cu2+, Zn2+ and Ni2+ ions. Ultrastructural study revealed many electron dense layers outside the cell-wall area that have not been observed earlier. PMID- 12744080 TI - Fatty acid composition of six freshwater wild cyanobacterial species. AB - Hydroxy, n-saturated, branched, dioic, and unsaturated fatty acids in six freshwater wild cyanobacteria (Chroococcus minutus, Lyngbya ceylanica, Merismopedia glauca, Nodularia sphaerocarpa, Nostoc linckia, and Synechococcus aeruginosus) collected from different lakes and springs of Israel have been identified by gas chromatography-mass spectrometry (GC-MS). PMID- 12744081 TI - Long-term fertilization affects the abundance of saprotrophic microfungi degrading resistant forms of soil organic matter. AB - The effect of mineral and organic fertilization on the occurrence of soil microorganisms was determined in a field experiment. The colony-forming unit counts of saprotrophic microfungi, when estimated on a silicate gel medium containing fulvic acid as a sole carbon source, increased significantly with increasing doses of mineral and organic fertilization. Partial correlation analysis indicated that, unlike bacteria and actinomycetes, microfungi utilizing fulvic acid were significantly associated with soil organic carbon. No significant effects on bacteria and microfungi counted on common microbiological media were observed but counts of actinomycetes increased in a manured soil extensively fertilized by a mineral fertilizer. Fulvic acid utilizing microfungi, which are associated with areas rich in organics, play possibly the main role in mineralization of resistant forms of soil organic matter. PMID- 12744082 TI - Competition among Bradyrhizobium strains for nodulation of green gram (Vigna radiata): use of dark-nodule strain. AB - The competitiveness of dual-strain inoculum of Bradyrhizobium strains S24 and GR4 was demonstrated for nodulation of green gram (Vigna radiata). Strain S24 formed pink nodules, GR4 produced visually distinguishable dark-brown nodules. When a mixture of these Bradyrhizobium strains was applied as inoculum, nodules of both pink and dark-brown types were formed on the same root. The strain GR4, which was less competitive than strain S24, was mutagenized with N-methyl-N'-nitro-N nitrosoguanidine to obtain pigment-diverse mutants and six selected mutants were screened for symbiotic parameters. One mutant produced pink nodules and appreciably increased plant dry mass. The competitive ability of this mutant lacking brown pigment was compared with that of strain S24 by using antibiotic resistance markers; it showed increased nodulation competitiveness than its parent strain GR4. The dark-brown nodule-phenotype could be useful in evaluating nodulation competitiveness of "cowpea miscellany" bradyrhizobia in soil where dark-brown nodule-forming strains are not indigenous. PMID- 12744083 TI - Water quality of some wells in Taiz City (Yemen Republic) and its surroundings. AB - Bacteriological and physicochemical parameters were determined in water samples collected from fifty-one producing wells used for human consumption in Taiz City (Yemen Republic) and its surroundings. Most water samples, especially from private wells, contained a high concentration of total coliforms which greatly exceeded the permissible limits recommended by the World Health Organization in 1996. Fecal coliforms were found in the majority of polluted sampled waters (65% of the total of wells examined). PMID- 12744084 TI - Standardized system for quantifying residual dirt on medical appliances cleansed in hospital washers--disinfectors: dirt detection by different methods. AB - An easy-to construct, easy-to-operate standardized system was developed for determining the residual biological contamination of surgical instruments, endoscopes and other medical appliances subjected to hospital cleansing and/or disinfection. It consists of standard-sized pieces of glass, metal or endoscope plastic--dirt carriers--either bare or enclosed in truncated Eppendorf caps to simulate hard-to-access conditions. The surface of the carriers is covered with model dirt simulating biological contamination and the carriers are then affixed to sturdy metal holders. Conventional model dirt were found to peel or flake off the carrier surface, lowering the precision of residual soil determination. A newly developed model dirt consisting of liver mash, lactose and sunflower oil and exhibiting low tendency to peel off surfaces was therefore used. The whole setup was subjected to chemical or enzymic cleansing programs at elevated temperature in hospital washer-disinfectors of two types, and the residual dirt after cleansing was determined by three methods. The method using toxicant-doped dirt that quenches the luminescence of an indicator bacterium Photobacterium phosphoreum gave satisfactory data under laboratory conditions but with hospital washed samples it exhibited excessive fluctuations caused by bacterium--dirt interactions and by physical influences. Both other methods gave better results but displayed some process sensitivity. The luciferin-luciferase-based ATP bioluminescence assay sometimes gave low or even negative dirt level values and showed a low effect of reduced dirt accessibility on cleansing of metal carriers. The Bradford protein assay showed about equal cleansing efficiency for both easily and poorly accessible carriers after enzymic cleansing. Our system can be used for determining low levels of residual contamination of medical appliances after cleansing/disinfection and assessing the efficiency of commercial washer disinfectors; its efficiency can be further increased by using a cleansing process-insensitive method for soil detection and quantification. PMID- 12744085 TI - Detection of Coxiella burnetii by PCR in mice after administration of live M-44 vaccine. AB - Original primers were prepared for the detection of coxiella burnetii by PCR for the amplification of a 752-bp fragment of dnaJ gene. Using these primers we observed the persistence of C. burnetii in different organs of mice after the administration of live Q-fever vaccine over a 7-months period. PMID- 12744086 TI - The presence of insertion elements IS861 and IS1548 in group B streptococci. AB - The presence of insertion elements (IS) IS861 and IS1548 in the collection of 211 Streptococcus agalactiae strains isolated from pregnant women and dairy cows was assayed. IS861 was found in 67 human strains (59%) and 36 bovine strains (37%), IS1548 in 13 human strains (12%) and 16 bovine strains (16%). Two combinations, IS861+ IS1548- and IS861- IS1548-, were widely distributed in both human and bovine strains. The copy number and the restriction fragment length polymorphism (RFLP) of the two IS were determined in human group B streptococcus (GBS) strains. A minimum of 8 copies of IS1548 were detected in GBS strains while the copy number of IS861 varied from 1 to 9. The number of different hybridizing patterns with IS861 and IS1548 probes was 9 and 6, respectively. These hybridization patterns were divided into several clusters. All strains with IS were also clustered according to pulsed field-gel electrophoresis (PFGE) patterns. A correlation was found between the results of PFGE- and IS-based clustering. PMID- 12744088 TI - [Deferred elective colonic resection in complicated acute diverticulitis]. AB - Twenty-three patients with acute diverticulitis complicated by pericolic or paracolic abscesses (Hinchey stage I-II) after a first phase of medical treatment were treated with deferred elective resection of the descending colon and sigmoid plus colorectal anastomosis performed on average 30 days after the onset of the acute episode. The pathologist's investigation of the surgical specimens demonstrated persistence of severe inflammatory lesions despite the apparently satisfactory clinical outcome. These data explain the frequent recurrences and indicate surgical treatment as being the only therapy capable of definitively resolving the condition. As compared with the emergency surgery performed by others, deferred elective resection makes it possible to operate on patients who, once the acute phase has been overcome, can have their hydroelectrolytic balance perfectly restored and be adequately monitored with treatment of associated diseases and perfect colon preparation. This strategy has allowed us to eliminate operative mortality and reduce the postoperative morbidity, both of which are significantly present in emergency surgical operations. Also the overall hospital stay in the two admissions, the interval between which can be reduced in ideal cases, does not significantly differ from that reported for emergency operations. PMID- 12744087 TI - Human tumor cells are selectively inhibited by colicins. AB - The activity in vitro of four types of colicins (A, E1, E3, U) against one human standard fibroblast line and against 11 human tumor-cell lines carrying defined mutations of the p53 gene was quantified by MTT (tetrazolium bromide) assay. Flow cytometry showed that the pore-forming colicins A, E1 and U affected the cell cycle of 5 of these cell lines. Colicins E3 and U did not show any distinct inhibitory effects on the cell lines, while colicins E1 and especially A inhibited the growth of all of them (with one exception concerning colicin E1). Colicin E1 inhibited the growth of the tumor lines by 17-40% and standard fibroblasts MRC5 by 11%. Colicin A exhibited a differentiated 16-56% inhibition, the growth of standard fibroblasts being inhibited by 36%. In three of the lines, colicins A and E1 increased the number of cells in the G1 phase (by 12-58%) and in apoptosis (by 7-58%). These results correlated with the data from sensitivity assays. Hence, the inhibitory effect of colicins on eukaryotic cells in cell selective, colicin-specific and can be considered to be cytotoxic. PMID- 12744089 TI - [Surgical treatment of para-esophageal hernia. Our experience]. AB - The authors report on 12 cases of para-oesophageal hernia surgically managed over the period from 1973 to 2001. They describe the etiopathogenetic factors responsible for the disease and the normal evolution of the clinical picture with the main complications, which are often the first manifestations. The main problems of surgical treatment are discussed with particular regard to the operative approach and to the usefulness of combining an antireflux procedure with the hiatoplasty. PMID- 12744091 TI - [Benign diseases of the thyroid: indications for surgical treatment and the current role of total thyroidectomy]. AB - Total thyroidectomy is the most popular surgical option in a wide range of indications for the treatment of benign disease of the thyroid. The preference for the procedure derives from a long period of observation and clinical experience dating as far back as the early 'seventies and confirms its safety and efficiency. Nevertheless, many doubts still exist as to the indications in the context of benign thyroid disease, as well as the incidence and seriousness of the complications which, though not frequent, are often invalidating. On the basis of the experience gained over the period from January 1994 to November 2002 in 697 patients undergoing surgery for benign disease of the thyroid, the authors analyse the indications for the various different therapeutic options and evaluate the results of total thyroidectomy in terms of therapeutic efficiency, relapse rates and complications. The latter are analysed on the basis of dividing the patients into 2 groups, one comprising 545 patients treated with total thyroidectomy as first-line treatment and the second consisting of 34 patients treated with total thyroidectomy for relapsing goitre after subtotal thyroidectomy. Comparison of the respective incidences of complications reveals a significant difference between the two groups of patients. Total thyroidectomy after subtotal thyroidectomy presented a significantly higher percentage of complications than initial total thyroidectomy. The authors come out in favour of total thyroidectomy with its low incidence of complications and the radical control of the disease it affords, reserving lobectomy-isthmectomy for selected cases of patients with monolateral disease which does not expose the patient to any risk of relapse. PMID- 12744090 TI - [Surgical treatment of breast cancer after neoadjuvant therapy]. AB - Primary chemotherapy represents a new therapeutic strategy that allows the implementation of conservative surgical treatment in locally advanced breast cancer or in cancers measuring > 3 cm. Of 127 patients with breast cancers measuring > 2.5 cm, 86 were treated with primary chemotherapy. This consisted in the administration of the ADM + TAX protocol in 28 cases and CNF in 58 cases. Three complete responses, 8 cases of stable disease and 75 partial responses were observed. Eleven radical mastectomies and 74 conservative treatments were performed. The chemotherapy protocol enabled us to convert 87% of the patients treated to conservative treatment, thus affording a treatment that offers a better quality of life despite the existence of a voluminous cancer and is of considerable psychological help in patients suffering from cancer of the breast. PMID- 12744092 TI - [Laparoscopic Nissen-Rossetti fundoplication: intermediate and long-term outcomes]. AB - The aim of this study was to report our personal experience with laparoscopic antireflux surgery using the Nissen-Rossetti fundoplication technique and to analyse the clinical and functional outcomes. From 1994 to 2002, 52 patients with gastro-oesophageal reflux disease underwent laparoscopic surgery after previously being submitted to endoscopy, gastro-oesophageal pH-metry and oesophageal manometry. A Nissen-Rossetti total fundoplication (short and floppy) was performed in all patients, even in those with defective peristalsis. The mortality rate was O and the mean operative time 72 minutes. Among the complications observed in the follow-up (3-100 months), dysphagia was the most frequent (11 patients, 21.1%); this was transient and invariably mild in 8 cases (15.4%), and persistent in 3 (5.7%), though again mild. The reflux symptoms were completely cured in 98.1% of the patients. This experience suggests that laparoscopic surgery is a safe and effective procedure for the treatment of gastro-oesophageal reflux disease. Precise selection of patients and adequate surgical technique are essential. PMID- 12744093 TI - Liver abscess due to acute cholecystitis. Report of five cases. AB - Acute cholecystitis is one of the most frequent causes of admissions to surgical departments. The development of liver abscesses is an uncommon and underrated complication of acute cholecystitis. In this study we report on our experience with the treatment of 5 cases of liver abscesses secondary to acute cholecystitis. All 5 cases were characterised by a lengthy period between the onset of acute cholecystitis symptoms and the subsequent diagnosis of a secondary liver abscess. In 4 out of 5 patients, admission for liver abscess occurred 12, 30, 50 and 120 days, respectively, after the acute cholecystitis episode. The liver abscesses were successfully treated with percutaneous drainage under US guidance (4 cases) and 4 patients underwent percutaneous cholecystostomy to treat the acute cholecystitis. After resolution of the acute phase, an elective cholecystectomy was performed in 4 out of 5 cases. Failure to diagnose acute cholecystitis at onset or inappropriate treatment of the condition could lead to the development of liver abscesses. PMID- 12744094 TI - [The learning curve in laparoscopic resections of the colon and rectum: results and considerations]. AB - A well-designed learning curve is essential for the success of laparoscopic colorectal surgery for cancer. The aim of this study was to evaluate the results and characteristics of the learning curve in laparoscopic colorectal surgery beginning with benign diseases and eventually going on to include colonic resections for cancer. A total of 60 laparoscopic resections were performed. In the first 33 cases only benign diseases (diverticular disease and polyps) were treated. The next 27 cases included resections for cancer, initially with the following exclusion criteria: obesity, previous abdominal surgery, emergency surgery for occlusion, voluminous tumours or infiltration of surrounding organs. Since January 2002 the only applicable exclusion criteria for laparoscopic resection have been emergency surgery for occlusion and invasion of adjacent organs. The following procedures were performed: 29 left hemicolectomies, 19 sigmoid resections, 7 segmentary resections, 3 abdomino-perineal resections and 2 right hemicolectomies. The conversion rate was 11.6%. The mean length of the segment removed was 21.5 cm. The mean number of lymph nodes harvested (for cancer) was 22.3. Major complications were observed in 3.3% and minor complications in 13.3%. The operative time decreased from a mean of 207 minutes to a mean of 170 minutes in the last group of 20 patients. Laparoscopic resections are safe and give the patient the opportunity to make a rapid recovery with less pain and a better outcome. We suggest performing laparoscopic colorectal resections initially for benign diseases (diverticular disease and polyps). This is needed in order to hone the technique. Resections for cancer can be undertaken only when the surgical team can guarantee an oncologically correct procedure in terms of lymphadenectomy, intraabdominal manipulation and extraction of the diseased segment from the abdomen. PMID- 12744095 TI - [Surgical treatment of complicated sigmoid diverticulitis: our experience]. AB - Traditionally, surgical sigmoid diverticular emergencies used to be treated in stages, but more recently there has been a trend towards definitive surgery with immediate resection plus anastomosis under certain conditions. The aim of this study was to define the morbidity and mortality of resection plus anastomosis with on-table antegrade irrigation and of the Hartmann procedure for complicated sigmoid diverticulitis in relation to the type of peritonitis and to the American Society of Anesthesiologists (ASA) grade of the patients. From April 1999 to April 2002, 38 emergency operations for complicated sigmoid diverticulitis were performed at the San Sebastiano Hospital in Caserta. Six patients underwent operations for obstructions and 32 for perforation (19 Hinchley stage III and 13 Hinchley stage IV). Surgical therapy for obstruction consisted in 4 resections plus anastomosis, 1 subtotal colectomy and 1 Hartmann procedure. Surgical therapy for perforation consisted in 14 resections plus anastomosis and 18 Hartmann procedures. There was 1 case (5%) of anastomotic dehiscence out of 19 primary anastomoses versus 2/19 surgical complications (10%) after the Hartmann procedure. The mortality amounted to 1 death out of 38 (2.6%) in a patient treated with the Hartmann procedure. Left-sided colonic obstruction should be treated by resection plus anastomosis or by subtotal colectomy for ASA II-III patients and by Hartmann's procedure for ASA IV-V patients. ASA II-III patients with localised or generalised non-faecal peritonitis should be treated by resection plus anastomosis, while a Hartmann procedure should be the reasonable option for generalised faecal peritonitis and for ASA IV-V patients with localised or generalised non-faecal peritonitis. PMID- 12744097 TI - [Gastrointestinal stromal tumors: current issues]. AB - Gastrointestinal stromal tumours are rare neoplasms originating from the connective tissue of the digestive tract and constitute most of the non epithelial primitive digestive tumours, with a percentage incidence of less than 1%. The term itself was first used in 1983 by Mazur and Clark to identify a heterogeneous group of tumours, all of them histologically characterised by hyperplastic fused cells, not necessarily leiomuscular ones, but even neural ones. The original stem cell has not been identified yet, but such tumours constantly present the expression of the surface antigens CD34 and CD117 which can be determined immunohisto-chemically. US endoscopy and fine needle aspiration with subsequent immunohistochemical analysis and study of c-kit gene mutation afford the best diagnostic accuracy. Current research is focused mainly on primary cells, probably the cells of Cajal, and on the study of the biological behaviour of gastrointestinal stromal tumours, which can be postulated by assessing several parameters, the most accurate of which seems to be the mitotic index. More recently, the possible therapeutic use of a tyrosine kinase inhibitor has been studied in tumours expressing the c-kit gene. Our experience is based on the histopathological and immunohistochemical study of 11 submucosal tumours (1 liposarcoma, 2 leiomiomas, and 8 gastrointestinal stromal tumours) out of a total of 75 submucosal tumours diagnosed. In the majority of cases a generic diagnosis of submucosal tumour was obtained with oesophagogastro-duodenoscopy and a surgical resection was then performed because of the symptoms. Only in 4 cases was the neoplasm found occasionally in the course of surgery for other reasons. The definitive diagnosis of gastrointestinal stromal tumour, as currently defined, was made only postoperatively by analysis of the histopathological and immunohistochemical findings. In all cases we evaluated the same parameters, i.e. actin, vimentine, S100, CD34, CD117, and Ki67, and confirmed constant high positivity for CD34 and above all for CD117. Even in the absence of unfavourable prognostic indicators, all patients are regularly followed-up. PMID- 12744096 TI - A prospective randomised study of local anaesthetic injection after multiple rubber band ligation of haemorrhoids. AB - One hundred and forty-two patients with second and third degree internal haemorrhoids were randomised to rubber band ligation only (n = 72) or rubber band ligation + local anaesthetic injection (n = 70). Pain was assessed by the patients at intervals of 6 hours and 1, 2, 3 and 4 days after banding. Other symptoms, complications, analgesic requirements and patient satisfaction were also recorded for 10 days following the treatment. There was a significant reduction in pain at 60 minutes and 6 hours after the procedure in the rubber band ligation plus local anaesthetic injection patients compared with the rubber band ligation only group (P < 0.05), but there was no reduction in pain when local anaesthetic was used compared with rubber band ligation only on days 1, 2, 3 and 4 days after ligation. On day 10 after banding, there was no difference between the two groups with respect to symptoms such as nausea, feeling of heaviness and/or tenesmus, fainting; complications, analgesic consumption or overall patient satisfaction. Bupivacaine injection after multiple rubber band ligation may be useful in reducing pain during the first 6 hours of the postbanding period. PMID- 12744098 TI - [Comparison of five synthetic absorbable suture materials in intestinal anastomosis: experimental study in rats]. AB - The purpose of this experimental study was to evaluate the efficacy of five synthetic absorbable suture materials in intestinal anastomoses in rats together with their interference with the normal physiopathological cicatrization process. The materials analysed were polyglycolic acid (Dexon), polygluconate (Maxon), polydioxanone (PDS), polygalactic acid (Vycril) and Glycomer 631 (Byosin). An anatomopathological study, performed in three groups of rats undergoing postmortem examinations after 6, 20 and 90 days showed that the least interference was caused by Glycomer 631, while polyglycolic acid and polygluconate yielded very good results though giving rise to a greater fibrous component. Polydioxanone and polygalactic acid yielded less satisfactory results. In addition, immunohistochemical techniques were applied to Glycomer 631. The evaluation of the surgical characteristics enabled us to divide the materials studied into two different groups, one in which characteristics such as smoothness and elasticity (Glycomer 631, polygluconate and polydioxanone) prevailed, and the second in which ease of knotting, stitch resistance, memory and lower cost (polyglycolic acid, polygalactic acid and Glycomer 63) prevailed. A general assessment showed better results for polyglycolic acid and Glycomer 631, while the other three materials presented slightly inferior performance, particularly polygalactic acid (inferior stitch resistance). PMID- 12744100 TI - Acute abdomen due to wandering spleen infarction: a case report. AB - We report a rare clinical case of acute abdomen due to partial infarction of a wandering spleen in the pelvis in a 60-year-old woman. The patient was suffering from stabbing pain in the external lower quadrant of the abdomen, irradiating back to the lumbosacral area, together with an unremitting feverish state (38 degrees C), sickness and constipation. After carrying out serological examinations, which revealed an increase in CPK and leukocytosis, ultrasonography and CT examinations were performed, revealing a mass in the left iliac cavity, which in all probability was a wandering spleen with an abnormally long pedicle and a dyshomogeneous lower area bearing witness to a splenic infarction. The patient was therefore submitted to surgery consisting in splenectomy after lysis of the adherences, which were plainly inflammatory. A wandering spleen, especially when infarcted, is a very rare clinical condition that may be congenital or acquired. Its presence can be confirmed by serological, ultrasonographical and CT examinations and must be suspected when there is no clearly defined acute abdomen. PMID- 12744099 TI - An unusual case of Cushing's syndrome due to ACTH-independent macronodular adrenal hyperplasia. AB - We report a case of Cushing's syndrome due to ACTH-independent macro-nodular adrenal hyperplasia (AIMAH). The patient, a 51-year-old man, had been suffering from poorly controlled arterial hypertension for the previous 6 years and he complained of progressive weight gain, gynecomastia and impotence. Physical examination revealed classic cushingoid features. Endocrine basal assessment showed increased urinary free cortisol (264-600 micrograms/24 hr). Urinary steroid profile showed an increase of total 17-hydroxycorticosteroids (17-OHCS: 23 mg/24 hr), with a threefold increase of tetrahydrocortisol (THF:9.66 mg/24 hr) and of tetrahydrocorticosterone (THB: 1.35 mg/24 hr). Tetrahydrodesossicortisol was only slightly elevated (THS:0.67 mg/24 hr) and tetrahydrodesossicorticosterone was at the inferior limit of the normal range (THDOC: 0.03 mg/24 hr). Total 17-ketosteroids were decreased (17-KS: 3 mg/24 hr). Plasma cortisol level was elevated and without circadian rhythm (26-29 micrograms/dl in the morning, 26-28 micrograms/dl at 24:00 h). DHEAs and free testosterone levels were significantly reduced (106 ng/dl and 3.9 pg/ml respectively). ACTH was undetectable and unresponsive to CRH. Both dexamethasone and octreotide failed to suppress plasma cortisol levels. Abdomen computed tomography scan demonstrated bilaterally enlarged multinodular adrenal glands. Cerebral magnetic resonance revealed no alteration of the pituitary gland. The patient underwent bilateral adrenalectomy. On macroscopic examination, adrenal glands were occupied by multiple yellow nodules and their compressive weight was 190 g, with left adrenal heavier than the right one (120 g and 70 g respectively). Histologically, nodular lesions were predominantly composed of large clear cells, with small foci of "hybrid" cells and adipose tissue metaplasia. Reticularis zone was atrophic. In the immediate post-operatory course pulmonary embolism occurred, despite prophylaxis with low molecular weight heparin. After having recovered from this complication, the patient showed progressive regression of cushingoid status. The findings of increased THF/THS and THB/THDOC ratios were in agreement with a relative hyperfunction of 11-beta hydroxylase "in vivo", which might have contributed to the hypercortisolism, in addition to the marked increase of secernent adrenal mass. PMID- 12744101 TI - [Nasopharyngeal carcinoma]. AB - The authors report the case of a 62-year-old woman who underwent a course of radiotherapy for an undifferentiated nasopharyngeal carcinoma. One year later the patient developed liver metastasis and underwent liver resection. The authors review the various aspect of this tumour, which is particularly frequent in the countries of South-East Asia but is exceptional in Europe and North America, focusing on the possibility of the operative management of the liver metastasis. PMID- 12744102 TI - Adrenal metastatic placental site trophoblastic tumor. Case report. AB - Placental site trophoblastic tumor is a rare disease. Since its first description in the literature only about 100 cases have been reported. Initially considered a benign pathology, its malignant potential with metastases was later recognized. About 30% of the cases described metastatised, thereby aggravating the prognosis. The therapy is surgical and in cases of metastatised disease has to be supplemented by chemotherapy. A case of placental site trophoblastic tumour with a single metastasis to the left adrenal gland treated with adrenalectomy and chemotherapy is reported. PMID- 12744103 TI - [Von Recklinghausen's disease and intestinal neurofibromatosis: a case report]. AB - We report the case of a patient with von Recklinghausen's disease, who was admitted with a diagnosis of acute abdomen due to small bowel neurofibromatosis. The patient was submitted to an abdominal CT scan that showed a homogeneous round lesion, with a regular margin, probably belonging to the small bowel and with the appearance of a benign lesion that probably caused an intestinal intussusception. The patient was submitted to a surgical procedure that mainly consisted in multiple small bowel resections. The histopathological examination confirmed the benign nature of the lesions. About one third of patients affected by von Recklinghausen's disease present involvement of the bowel, but only 5% of them are symptomatic. The intestinal tumours are usually neurofibromas and are mainly localized in the jejunum. However, there have also been reports of stromal, nervous and endocrine tumours and even other tumours not belonging to these categories, including adenocarcinoma. The overall incidence of intestinal malignancy in patients with von Reckilnghausen's disease is about 10%. The surgical operation, as well as the histopathological and immunochemical examination of the intestinal lesions are of crucial importance for the treatment of the complications of intestinal neurofibromatosis and for the treatment and diagnosis of malignancy. PMID- 12744104 TI - Aneurysms of the gastroduodenal artery: a case report. AB - Aneurysms of the gastroduodenal artery are the rarest of all splanchnic artery aneurysms. A case of gastroduodenal artery aneurysm diagnosed preoperatively and successfully managed surgically is described. A 68-year-old woman was admitted to our institution as a result of recurrent epigastric pain. Arteriography of the coeliac trunk and of the upper mesenteric artery showed a coeliac axis occluded at the origin and a voluminous round aneurysmatic formation of the gastroduodenal artery measuring approx. 3 cm. Such lesions may be asymptomatic and initially pass unnoticed or may appear only on rupturing with a profile of haemoperitoneum. The preoperative diagnosis is difficult: in many cases only non-specific signs such as pain, jaundice or upper digestive tract haemorrhage are present. Selective arteriography, in this case, permitted the definitive diagnosis of a gastroduodenal aneurysm, as well as establishing its size and morphology and the vascular pattern of the lesions, all of which was of great assistance in selecting the surgical strategies to be adopted. The operation consisted in the proximal and distal ligation of the gastroduodenal artery and in resection of the aneurysm after preparation of the portal vein, the common hepatic artery and the VBP. We maintain that, because of the risk of rupture of the aneurysm, which so often has a fatal profile, an aggressive surgical approach is both justifiable and advisable. PMID- 12744105 TI - [Small intestinal polyposis in Peutz-Jeghers syndrome: combined treatment with surgery and intra-operative endoscopy]. AB - Polyps occur throughout the gastrointestinal tract in Peutz-Jeghers syndrome, but the most serious problems are encountered in the management of small bowel polyposis. We report here on a case of Peutz-Jeghers syndrome admitted to hospital for intestinal obstruction and anaemia. The patient was submitted to colonoscopy, oesophagogastro-duodenoscopy and small bowel enema. At laparotomy, multiple intussusceptions were found and we conducted a combined surgical endoscopic approach. Most of the polyps were identified and removed endoscopically (snare polypectomy). Five enterotomies were performed to remove 18 very large polyps (> 3 cm). Finally, a limited portion of the jejunal tract (20 cm) was resected owing to the presence of multiple, large, obstructive polyps. None of the polyps showed cancerous transformation. The shortcomings of the traditional surgical approach include repeated small bowel resections and often early reoperation to manage complications caused by polyps missed at the time of previous surgery. If surgical intervention is required, intraoperative endoscopy is always indicated. Conservative surgical management, the role of intraoperative endoscopy, planned medical follow-up and the need for a national registration system are stressed. PMID- 12744106 TI - Hydatid disease in a very unusual location: the adrenal gland. A case report. AB - Hydatid cysts are most often located in the liver and lungs (55-90%), while their location in the adrenal gland is very rare (0.5%). We observed a patient with a hydatid cyst in the adrenal gland, the diagnosis of which was incidental during ultrasonography and computed tomography. The experience acquired in Sicily where hydatid disease is endemic makes serological tests unnecessary, also because they often require a lengthy waiting period and are expensive. The patient had undergone surgery for the treatment of hepatic hydatid cysts. The adrenal localization may be explained as a consequence of secondary dissemination via the blood stream. The cyst developed inside the gland and caused atrophy of the glandular tissue. The surgical treatment called for adrenalectomy as the adrenal gland was entirely occupied by the cyst. PMID- 12744107 TI - Pancreatic pseudocyst caused by extracorporeal shock wave lithotripsy for right renal pelvic calculi. AB - A 39-year-old white man with a history of right renal pelvic stones treated 1 year before by extracorporeal shock wave lithotripsy, but with no history of major surgery, alcoholism, pancreatitis, hyperparathyroidism or trauma, was admitted, suffering from an abdominal mass. Abdominal and pelvic computed tomography revealed an enlarged pancreatic head (9 cm in transverse diameter) with, inside it, a heterogeneous, cyst-like structure, measuring 7 cm in diameter. It was suspected that this lesion was a cystic neoplasm and the patient underwent a proximal pancreaticoduodenectomy and a cholecystectomy. After the operation, the microscopic findings ruled out the presence of a neoplasm and we were obliged to reconsider the case. Speculating as to the possible role of past extracorporeal shock wave lithotripsy in determining the pancreatic pseudocyst, it was found that damage to the intra-abdominal organs during extracorporeal shock wave lithotripsy has been mentioned in published series, but it was also noted that this case seemed to differ from the other published cases, where cirrhosis and thrombocytopenia, gallbladder stones, or adhesions between the pancreas and surrounding tissue caused by laparotomy were considered the causes of the lithotripsy complications. We suggest a direct traumatic disruption of the pancreas as a result of the extracorporeal shock wave lithotripsy and conclude that the post-lithotripsy follow-up should include periodic ultrasonographic investigation of the pancreas and serum amylase level determinations in order to diagnose pancreatic complications, if any, and plan the correct treatment. PMID- 12744108 TI - Pancreatic rupture following a bull-head caused blunt abdominal trauma. AB - A 33-year-old man was admitted to our hospital with clinical and instrumental diagnostic evidence (CT) of haemoperitoneum following a blunt abdominal trauma due to being butted by a bull. A full-thickness rupture of the pancreas, at the level of the isthmus, was found at laparotomy. The emergency procedure consisted in evacuation of the haemoperitoneum, haemostasis and suture of the pancreatic lesion plus drainage of the lesser sac. A high-output (800 ml/day) post-traumatic pancreatic fistula developed in the following days, with the need for a second operation. In the presence of a large pancreatic defect extensively communicating with the Wirsung duct, a singular surgical procedure was adopted in the form of a Roux-en-Y pancreaticojejunostomy with catheterisation of the pancreatic fistula using a small disposable silicone catheter. The post-operative course was uneventful and healing of the pancreatic fistula was documented at CT scan 39 days after the operation. PMID- 12744109 TI - [A rare case of digestive tract hemorrhage of extraluminal origin after total gastrectomy]. AB - The authors report on a case of digestive bleeding (melaena and enterorrhagia) in a patient undergoing total gastrectomy for gastric cancer and later splenectomy for subcapsular haematoma in a different hospital. The source of bleeding was not intraluminal; the bleeding arose from double erosion of the gastroduodenal artery in the tract above the anterior surface of the pancreas, close to the dehiscent duodenal stump. The blood flowed mainly into the enteric district through the open stump thus causing the clinical signs described. The diagnosis was made during an emergency surgical operation for haemorrhagic shock. The patient underwent haemostasis with two stitches on the gastroduodenal artery, external drainage of the duodenum with a Petzer tube, laparostomy of the infected area and ileostomy. After three months he had completely recovered. PMID- 12744110 TI - Major vessel injuries during laparoscopic cholecystectomy: a case report. AB - The aim of the study was to investigate major vascular injury during laparoscopic cholecystectomy, which is a rare but potentially lethal complication if not recognized and treated swiftly, usually secondary to placement of the Veress needle or trocar. We report on our experience with a 35-year-old woman presenting an aortic laceration of the anterior wall and a puncture wound penetrating the anterior wall of the inferior vena cava. Repair of the injuries was done using a 5-0 Prolene suture mounted on pledgets. In addition, it was necessary to suture three mesenteric vascular injuries in the posterior peritoneum with Vicryl 2-0. The estimated blood loss was 1300 cc. The patient did well and was discharged on postoperative day 13. The distal abdominal aorta, inferior vena cava and large pelvic vessels are particularly prone to injury during laparoscopic cholecystectomy. In most cases, the vascular defect can be closed by direct suturing. Rarely is it necessary to reconstruct the injured vessel with an alloplastic prosthetic patch or a prosthetic implant. PMID- 12744111 TI - [Laparoscopic diagnosis of gallbladder agenesis: clinical case]. AB - Congenital absence of the gallbladder is a rare anomaly. We report a case of agenesis of the gallbladder in a 45-year-old woman incorrectly diagnosed as cholelithiasis on preoperative ultrasound. The diagnosis of agenesis was confirmed laparoscopically with laparoscopic exploration, intraoperative ultrasound and cholangiography. Formal laparotomy was avoided. Difficulties in management during laparoscopic surgery are discussed. A review of the literature is presented. PMID- 12744112 TI - [Ultrasonic scalpel in thyroid surgery]. AB - In order to evaluate the harmonic scalpel during "open" thyroid surgery, a prospective randomized study was performed using the device versus the traditional procedure. One hundred and forty-two multinodular goiter patients were selected for total thyroidectomy and subdivided into two gender- and age matched groups. In the first group of 72 patients, the surgical procedure was performed with the US scalpel without using any electrical device or vascular ligatures; in the second group of 70 patients, the traditional technique was employed. No statistically significant differences were recorded with regard to permanent inferior laryngeal nerve palsy or hypoparathyroidism rates. Use of the harmonic scalpel was associated with a higher incidence of transient nerve injury (P < or = 0.005, 9.7% vs 1.4%), but this difference disappeared with increasing experience with the harmonic scalpel. Using the US device significantly improves surgical duration (87.5% < 1 h) and blood loss (P < or = 0.005). PMID- 12744113 TI - Use of the transcystic guidewire in single-intervention treatment for cholecysto choledocholithiasis. A case report and technical notes. AB - Videolaparoendoscopic treatment of choledocholithiasis in a single stage is an important option for this disease. We currently adopt this approach to choledocholithiasis in our department. We report here the case of a woman with stones in the biliary tract and gallbladder. After videolaparoscopic cholecystectomy we performed a transcystic cholangiography. A guidewire was used to show Vater's papilla during endoscopic papillosphinterotomy, because this was in a duodenal diverticulum that made it impossible to cannulate the papilla. We propose this method in all those cases in which, for anatomical reasons, the papilla cannot be easily cannulated. PMID- 12744114 TI - Prospective randomised study of cephalic vein cut-down versus subclavian vein puncture. Technique in the implantation of subcutaneous venous access devices. PMID- 12744117 TI - See the signs. PMID- 12744116 TI - Wings of change. PMID- 12744115 TI - Looking into alternatives. PMID- 12744119 TI - Money is the answer. PMID- 12744118 TI - Push your IQ up the scale. PMID- 12744120 TI - It's your move. PMID- 12744121 TI - Constipation care. PMID- 12744123 TI - Medical humanities in nurse education. AB - The author describes the way in which nursing knowledge is developing through medical humanities. This is the study of medical themes and concerns in fictional representations of illness and health, to understand the effect of illness on individuals and society. Thus, the study of medical humanities shows how the humanities, such as literature and philosophy, can enhance nurse education. PMID- 12744122 TI - Evidence-based patient handling: systematic review. AB - BACKGROUND: This article summarises the findings of an international systematic review of patient handling. More than 880 papers were read, critically appraised and allocated a quality rating score. The evidence supports the provision of a minimal set of equipment for moving and handling patients, and multifactor intervention strategies based on risk assessment rather than those predominantly based on technique training. CONCLUSION: It is recommended that healthcare providers should review their current equipment provision and approach to managing risks and injuries associated with patient handling activities in the context of this research evidence. PMID- 12744124 TI - Prevention and control of viral hepatitis. AB - Viral hepatitis is a common disease caused by various hepatotropic viruses. An awareness of the characteristics and transmission patterns of these viruses will help nurses to understand the diverse clinical outcomes of infection and develop effective patient education and infection prevention and control strategies. PMID- 12744125 TI - Checks and balances. PMID- 12744127 TI - [Roentgenosurgery for aortic coarctation and open arterial duct with intravascular ultrasonic examination]. AB - Forty-five patients (32 with open arterial duct and 13 with aortic coarctation) were examined. Forty-nine intravascular ultrasonic examinations were performed, qualitative and quantitative analysis of 139 vascular segments was carried out. Despite the fact that mean morphometric indices of open arterial duct and aorta in angiography and intravascular ultrasonic examination had no significant difference, real confidence of the o data was seen only in 13.3% cases for open arterial duct an in 16.7% cases--for aortic coarctation. In 37.5 and 50% patients, respectively, difference in the dimensions was so significant that dictated choice of instruments with difference size. Roentgenoendovascular surgeries were performed in 23 patients with open arterial duct and in 12--with aortic coarctation. Choice of instruments parameters based on data of intravascular ultrasonic examination provided safer and more efficient procedures. PMID- 12744126 TI - [Surgical treatment of resistant ascites in patients with portal hypertension]. AB - Surgical methods of treatment of diureticoresistant ascites in 198 patients with portal hypertension are analyzed: portocaval bypass (n = 16), lymphovenous anastomosis (n = 64), ductolysis (n = 24), and peritoneovenous bypass (n = 94). The most stable positive results was achieved after portocaval bypass, but it can be performed only in a small part of patients. Peritoneovenous bypass and lymphovenous anastomosis demonstrated shorter effect but they had less risk and were better tolerated than portocaval bypass. Efficacy of these surgeries depends not only on accurate indications and contraindications but also on functional resource of the liver and organism's reserves. Following these conditions and individual approach to choice of surgical methods of resistant ascites' treatment may improve quality of life and lifespan of patients with portal hypertension. PMID- 12744128 TI - [Low-intensive laser irradiation in combined treatment of lower limbs atherosclerotic lesions]. AB - Seventy-one subjects entered this study. The control group consisted of 12 healthy subjects, the comparative group included 15 patients who received standard therapy of vascular diseases but without physiotherapy. The study group consisted of 44 patients whose treatment was supplemented with laser irradiation. Angiography, ultrasonic dopplerography, laser flowmetry, oxygenometry were applied for control of treatment efficacy. Regional ischemia was evaluated with detection of pO2 of foot. LT increased oxygenation of foot soft tissues in patients with low primary pO2 and decreased in ones with higher. As a result the number of patients with low pO2 (0 < pO2 < 20) decreased from 13.7 to 4.5%, with middle pO2 (20 < pO2 < 40) increased from 27.3 to 50.0%, with high pO2 (pO2 = 40) decreased from 59.0 to 45.5%. Redistribution in favor of 20 < pO2 < 40 is regarded as normalizing effect of LT. It is concluded that LT increases oxygenation of foot soft tissues in patients with low primary pO2 and decreased in ones with higher. PMID- 12744129 TI - [Diagnosis and treatment of advanced stomach cancer]. AB - 800 patients with stomach cancer were operated in Saratov Military Medical Institute from 1990 to 2000. Results of radical surgery in advanced stomach cancer are presented. The most frequent postoperative complications and causes of lethal outcomes are described. Treatment and diagnostic algorithm in stomach cancer is proposed to optimize work of general surgery units. PMID- 12744130 TI - [A two-layer perforated synthetic polytetrafluoroethylene plate in the treatment of inguinal hernia]. PMID- 12744131 TI - [Surgical policy in nonspecific ulcerative colitis]. AB - Results of surgical treatment of 101 patients with nonspecific ulcerative colitis are analyzed. In 73 (72.3%) patients variants of abdomino-anal coloproctectomy were performed, 22 of them underwent total coloproctectomy with terminal ileo- or colostoma creation. In 21 (20.8%) patients who had no inflammatory-ulcerative process in the rectum resection of the affected part of the colon with ileo--or colorectal anastomosis was performed, in 8 cases suturing device AKA-2 was used. In 7 (6.9%) patients who had undergone total coloproctectomy S-type intestinal reservoir and reservoir-anal anastomosis were created. Rate of postoperative complications was 16.8%, lethality--3.0%. 6 months after creation of reservoir anal anastomosis nearly complete recovery of anal sphincter function was seen that permits to regard this surgery as a method of choice in the treatment of patients with total ulcerative colitis. PMID- 12744132 TI - [High-frequency insufflation of drug aerosol into abdominal cavity in combined treatment of acute general peritonitis]. AB - Method of treatment of abdominal cavity in acute general peritonitis--high frequency insufflation of drug aerosol into abdominal cavity--was developed, tested in animals and used in 126 patients. Results of treatment were compared with a standard method of surgical practice--peritoneal lavage. It is demonstrated that the method of high-frequency insufflation promotes a more favorable course of peritonitis, decreases in the number of purulent-septic complications and reduces time and cost of treatment. PMID- 12744133 TI - [Postoperative complications in patients with peritonitis]. AB - One thousand three hundred and ten patients with peritonitis have undergone surgery: local limited peritonitis was in 230 patients, local unlimited--in 342, general--in 738 patients. After surgery suppuration of the wound was seen in 92 (7.1%) patients, phlegmons of the abdominal wall--in 16 (1.3%), progressive peritonitis--in 40 (3.1%), abscesses of abdominal cavity--in 13 (1.0%), eventration--in 19 (1.5%), adhesive intestinal obstruction--in 17 (1.3%). Laparoscopy was used for diagnosis of intraabdominal complications in 47 patients. This allowed to avoid unjustified laparotomies in 20 patients. Application of laparoscopy is limited by degree of bacterial contamination of peritoneal exudates. Early adhesive intestinal obstruction in 7 patients was treated with endoscopic procedure. Sonography is effective in diagnosis of intraabdominal complications, detection of site of safe puncture before postoperative laparoscopy and drainage of abscesses. PMID- 12744134 TI - [Use of lectin for detection of changes of erythrocytic osmotic resistance and deformity in acute advanced peritonitis]. AB - Thirty-one patients with acute general peritonitis were studied. Lectin (LPM) demonstrated an additional peak on kinetic curves of erythrocytic osmolysis in patients with acute general peritonitis. Deformity of erythrocytes had a tendency to decrease (p > 0.05), but in the presence of LPM this difference increased sharply (p < 0.001). On day 7 after surgery in case of a favorable outcome the peak decreased or disappeared, deformity of erythrocytes also disappeared. It is important for diagnosis and prognosis. PMID- 12744135 TI - [Ways of raising efficacy of nasointestinal drainage in patients with intestinal obstruction and general peritonitis]. AB - Expediency of programmed nasointestinal decompression with gastrointestinal aspirator GOMCO (USA) in patients with intestinal obstruction and general peritonitis in parameters 90 and 120 mm Hg was substantiated. If introduction of the gastric tube into the jejunum is impossible (in patients with adhesive obliteration of higher part of abdominal cavity, severe deformation or obstruction of pyloroduodenal canal) but in absolute indications for long-term intestinal drainage it is expediently to perform extraduodenal intubation of the small intestine by means of creation of posterior minigastrojejunoanastomosis on short loop behind colon. PMID- 12744136 TI - [Etiology, pathogenesis, classification and surgical treatment of diabetic foot syndrome]. AB - Basic points of ethiology and pathogenesis of diabetic foot syndrome (DFS) are given. Classification of DFS is proposed as regards to clinical forms, stages and localization of the process. Algorithm of treatment of each clinical form of DFS including surgical methods is substantiated. Indications for organo-saving surgeries on the foot, and also for high amputation of the leg are determined clearly. Results of treatment of 484 patients with DFS are analyzed. This algorithm can reduce number of high amputations 2 times. It is noted that early hospitalization of patients with initial stages of the disease, organization of curative process depending on DFS clinical form permits to improve results of FDS treatment. PMID- 12744137 TI - [Features of intestinal invagination in children older than one year of age]. AB - Results of treatment of 368 children with intestinal invagination are analyzed for 1976-2001. Features of intestinal invagination are determined. One of the main features is increase to 50% in the number of patients with this disease older than one year. Invagination in these patients progresses more mildly and has a more benigh course leading to diagnostic mistakes before hospitalization in 85.8% cases and ones in surgical units--in 3.5%. Efficacy of conservative treatment of invagination in children older than one year (95.8%) and of recurrence of the disease (100%) is demonstrated. Conservative desinvagination is the method of choice in the treatment of intestinal invagination regardless of age. PMID- 12744138 TI - [Carbapenem antibiotic ertapenem in the treatment of extrahospital intraabdominal infections]. PMID- 12744139 TI - [About classification of anterior resections of the rectum]. AB - Results of anterior resections of rectum are analyzed. It is demonstrated that results of treatment and surgical policy depend on level of resection and anastomosis creation. Own classification of anterior resections of rectum are developed. This classification may be used as the base of coherent treatment policy. PMID- 12744140 TI - [Cystic transformation of vestigial ureter in a child]. PMID- 12744142 TI - [Russian priority in development of knowledge about etiology and pathogenesis of gastric and duodenal ulcers]. PMID- 12744141 TI - [Molecular-genetic factors in cancer of the pancreas]. PMID- 12744143 TI - [German surgeon Ferdinand Sauerbruch]. PMID- 12744145 TI - Altruism and selfishness. AB - Many situations in human life present choices between (a) narrowly preferred particular alternatives and (b) narrowly less preferred (or aversive) particular alternatives that nevertheless form part of highly preferred abstract behavioral patterns. Such alternatives characterize problems of self-control. For example, at any given moment, a person may accept alcoholic drinks yet also prefer being sober to being drunk over the next few days. Other situations present choices between (a) alternatives beneficial to an individual and (b) alternatives that are less beneficial (or harmful) to the individual that would nevertheless be beneficial if chosen by many individuals. Such alternatives characterize problems of social cooperation; choices of the latter alternative are generally considered to be altruistic. Altruism, like self-control, is a valuable temporally-extended pattern of behavior. Like self-control, altruism may be learned and maintained over an individual's lifetime. It needs no special inherited mechanism. Individual acts of altruism, each of which may be of no benefit (or of possible harm) to the actor, may nevertheless be beneficial when repeated over time. However, because each selfish decision is individually preferred to each altruistic decision, people can benefit from altruistic behavior only when they are committed to an altruistic pattern of acts and refuse to make decisions on a case-by-case basis. PMID- 12744144 TI - Mental imagery: in search of a theory. AB - It is generally accepted that there is something special about reasoning by using mental images. The question of how it is special, however, has never been satisfactorily spelled out, despite more than thirty years of research in the post-behaviorist tradition. This article considers some of the general motivation for the assumption that entertaining mental images involves inspecting a picture like object. It sets out a distinction between phenomena attributable to the nature of mind to what is called the cognitive architecture, and ones that are attributable to tacit knowledge used to simulate what would happen in a visual situation. With this distinction in mind, the paper then considers in detail the widely held assumption that in some important sense images are spatially displayed or are depictive, and that examining images uses the same mechanisms that are deployed in visual perception. I argue that the assumption of the spatial or depictive nature of images is only explanatory if taken literally, as a claim about how images are physically instantiated in the brain, and that the literal view fails for a number of empirical reasons--for example, because of the cognitive penetrability of the phenomena cited in its favor. Similarly, while it is arguably the case that imagery and vision involve some of the same mechanisms, this tells us very little about the nature of mental imagery and does not support claims about the pictorial nature of mental images. Finally, I consider whether recent neuroscience evidence clarifies the debate over the nature of mental images. I claim that when such questions as whether images are depictive or spatial are formulated more clearly, the evidence does not provide support for the picture-theory over a symbol-structure theory of mental imagery. Even if all the empirical claims were true, they do not warrant the conclusion that many people have drawn from them: that mental images are depictive or are displayed in some (possibly cortical) space. Such a conclusion is incompatible with what is known about how images function in thought. We are then left with the provisional counterintuitive conclusion that the available evidence does not support rejection of what I call the "null hypothesis"; namely, that reasoning with mental images involves the same form of representation and the same processes as that of reasoning in general, except that the content or subject matter of thoughts experienced as images includes information about how things would look. PMID- 12744146 TI - Clinical and sonographic estimations of uterine weight. AB - Over a 36 month period, all hysterectomies with clinical assessment, sonographic measurement of uterine length and estimated uterine weight were reviewed. Accuracy of these methods was ascertained by: 1) mean error, 2) mean standardized error, and 3) percentage of estimates within 10% of the actual weight. Among 210 hysterectomies, the mean (+/- standard deviation) of uterine weight was 514.9 gm +/- 647.0 gm (range 54-6700 gm) with 135 specimens (64.3%) being 500 gm and 75 > 500 gm. The receiver-operating curves for clinical assessment (0.82 +/- 0.03), uterine length (0.87 +/- 0.03) and Linear Regression Equation (0.82 +/- 0.04) are not significantly different (P > 0.05), having similar abilities to differentiate uterine weight. The clinical estimate of uterine weight is as accurate as ultrasound in determining a uterus weighing > 500 gm but all three methods poorly predict actual uterine weight. PMID- 12744147 TI - Twenty years and counting. PMID- 12744148 TI - Looking back: two cases of abscess of the liver, treated by incision and free drainage. PMID- 12744150 TI - [Influence of different contact ways and extracting conditions on the hemolytic effect of biomaterials]. AB - This assay was aimed to evaluate the influence of different contact ways and extracting conditions on the hemolytic effect of biomaterials. Using direct contact method and extract contact method, we assessed the hemolytic effect of PDLLA and PVC. The extracting conditions included: 37 degrees C 24 h, 37 degrees C 72 h, 37 degrees C 120 h, 50 degrees C 72 h, and 70 degrees C 24 h. After the material or extract had been in contact with the diluted blood of rabbit for certain times, the hemolysis rate was calculated. The results for PDLLA showed there were some differences between direct contact and extract contact at 37 degrees C for different extraction time (P < 0.05), but the hemolysis rates, lower than 5%, were in accord with the requirements of medical devices. However, under the condition of 50 degrees C and 70 degrees C, there were significant differences when extract contact method was compared with direct method (P < 0.01). For PVC, there was no statistically significant difference under all conditions (P > 0.05). Our conclusions: (1) Under the extracting condition of 37 degrees C from 24 h to 120 h, the soluble part of PDLLA and PVC that might influence erythrocyte did not dissolve considerably. (2) Under the extracting condition of 50 degrees C and 70 degrees C, the hemolysis rate may remarkably vary with the chemical characteristics of tested materials; (3) As to an unknown material, it is advisable to adopt two methods at the same time, one for direct contact and the other for extracontact. Thus the hemolytic effect of biomaterials can be evaluated from physical and chemical angles. (4) In case that the chemical property of the sample can endure the test, the extracting condition at 50 degrees C and 70 degrees C may be of benefit to assessing the hemolytic effect of biomaterials. (5) The extract contact method as a supplemental test of direct contact method is of realistic significance. PMID- 12744149 TI - [An investigation of HAP/organic polymer composite coatings prepared by electrochemical co-deposition technique]. AB - An electrochemical co-deposition technique has been developed to prepare a hydroxyapatite (HAP)/organic polymer composite coatings on Ti surface as new biomaterial of hard tissue. The composite coating of organic polymer and calcium phosphate is formed by adding a water soluble polymer of the ethylene series to NH4H2PO4-Ca (NO3)2 solution when conducting an appropriate electrochemical co deposition experiment. The XRD, SEM, XPS, SIMS and nano indent measurements were performed to characterize the morphology, composition, structure and surface stiffness of the composite coating. It was found that the morphology and surface hardness of the coatings showed a remarkable modification when introducing a minor polymer to HAP coating, and the bonding force between the coating and metal substrate was distinctly increased. The incorporation of minor organic polymer into the HAP compound at molecular level will improve the mechanical properties and morphology of the composite coatings, and this may be helpful to raising its bio-activity. PMID- 12744151 TI - [Elimination of methylene blue in plasma by use of adsorbents]. AB - The adsorption of methylene blue (MB) in plasma on cross-linked agarose beads entrapped activated charcoal (CAAC-I), cross-linked agarose coated activated charcoal (CAAC-II) and cross-linked agar beads entrapped attapulgite (CAA) was tested in this study. These adsorbents are intended to be applied to blood purification for eliminating MB from virucidal MB-phototrated plasma units. The experimental results indicated that the adsorption of MB in plasma on CAAC-I and CAA was quick and efficacious, but it was slower on CAAC-II. The relationship between the adsorption time and the adsorption rate was examined with reference to the plasma with a specific concentration of MB flowing over a certain volume of CAA. A flow rate of about 1 ml.min-1 through 10 ml CAA at 25 degrees C could eliminate more than 95% of the MB in 200 ml human plasma (1 mumol.L-1). PMID- 12744152 TI - [Study on the cytocompatibility of biodegradable poly(epsilon-caprolactone) microspheres in vitro]. AB - Poly(epsilon-caprolactone) microspheres were fabricated with an average particle size of 5.08 +/- 0.23 microns. The effect of poly(epsilon-caprolactone) microspheres on apoptosis and cell cycle of fibroblast was studied with flow cytometry. The data obtained clearly indicated that poly(epsilon-caprolactone) microspheres purified in different ways showed different cytocompatibility. The well purified microspheres have good cytocompatibility. PMID- 12744153 TI - [Two sorts of problems on drug controlled release from swellable polymer]. AB - The mechanism of drug release from swellable polymer was studied. The model which can explain the drug release from glassy polymer was developed by coupling drug diffusion equation and solvent diffusion equation. The effect of viscoelastic stress of polymer was also considered. The Deborah number and swelling interface number were used to describe the mechanism of drug release, especially relax controlled mechanism. PMID- 12744154 TI - [A study of bone-like apatite formation on calcium phosphate ceramics in different kinds of animals in vivo]. AB - Bone-like apatite formation on the surface of calcium phosphate ceramics has been believed to be necessary for new bone to grow on the ceramics and to be related to the osteoinductivity of the material. The research of bone-like apatite formation is a great help to understanding the mechanism of osteoinduction. Synthetic porous calcium phosphate ceramics (HA/TCP = 70/30) were implanted intramuscularly in pigs, dogs, rabbits and rats to make a comparative study of the bone-like apatite formation onto the porous HA/TCP ceramics in different animals. Specimens were harvested at 14 days after implantation. Samples were detected for the surface morphology with SEM. The chemical composition of the sample surface after implantation was analyzed with reflection infrared (R-IR). Obvious bone-like apatite formation could be detected in the sections of porous specimens harvested from all animals after 14 days intramuscular implantation. Crystal deposition could be only observed on the surface of the concave regions of the samples collected from dogs, rabbits and rat. On the contrary, evenly distributed flake-shaped crystal could be found on the pore surface and also on the outer surface of the materials implanted in pigs. The morphology of bone-like apatite in pigs was different from that in the others animals. Bone-like apatite was not observed in dense specimen implanted intramuscularly. Bone-like apatite formed faster on specimens implanted in rabbit than that in other animals. This formation sequence is different from the sequence of osteoinductivity of biphasic calcium phosphate ceramics implanted in these animals. The results demonstrated that the formation of bone-like apatite on materials is a prerequisite condition to their osteoinduction but other factors also play important roles in osteoinduction. PMID- 12744155 TI - [Preparation of complex chitosan microcapsule and its application in controlled release of vitamin D2]. AB - In this work a system which consists of chitosan (CS) microcores entrapped within enteric polymer is presented. Vitamin D2, used as a model drug, was efficiently entrapped within CS microcores using spray-drying and then microencapsulated into ethylic cellulose(EC). The morphology and release properties of microcapsules were tested. The influential factors of preparation conditions included molecular weight of chitosan, concentration of chitosan solution, concentration of acetic acid, loading of vitamin D2 were discussed. The results of in vitro release studies showed that the microcapsules prepared in this article could realize sustained release in intestine. PMID- 12744157 TI - [Mechanics analysis of fracture of orthodontic wires]. AB - Fracture problem of orthodontic wires was discussed in this paper. The calculation formulae of bending stress and tensile stress were obtained. All main factors that affect bending stress and tensile stress of orthodontic wires were analyzed and discussed. It was concluded that the main causes of fracture of orthodontic wires were fatigue and static disruption. Some improving proposals for preventing fracture of orthodontic wires were put forward. PMID- 12744156 TI - [A biomechanical model for simulating the deformation of a leukocyte adhered to the surface of a blood vessel under steady shear flow]. AB - The adhesion of leukocytes to substrate is an important biomedical engineering problem and has drawn extensive research. In this study, we have proposed a compound drop model to simulate a leukocyte with a nucleus adhered to the surface of a blood vessel under steady shear flow. A two-dimensional computational fluid dynamics (CFD) is conducted to determine the local distribution of pressure on the surface of the adherent model cell. By introducing the parameter of deformation index (DI), we have investigated the deformation of the model cell and it's nucleus under controlled conditions. Our numerical results show that: (1) the model cell is capable of deformation with the increase of initial contact angle, capillary number, and Reynolds number, and that the cytoplasm is more deformable while the nucleus is more capable of resisting external imposed shear flow; (2) the model cell is not able to deform infinitely with the increase of external shear flow because the deformation index reaches a maximum; (3) pressure distribution confirms that there exists a region downstream of the cell, which produces high pressure to retard continuous deformation and provide a positive lift force on the cell. Our results of nucleus deformation may help to develop a better understanding of how leukocytes transduce external mechanical signal like shear stress into nucleus. PMID- 12744158 TI - [Experimental study on the viscoelasticity of aorta ascendens]. AB - We carried out the stress relaxation and creep experiments on the aorta ascendens of five fresh dead bodies of normal chinese men who had died from impetuous trauma on the head. The stress relaxation and creep data and curves of the aorta ascendens were obtained. Then regression analysis was performed to get the reduced stress relaxation and creep functions. Finally, we analyzed and discussed the results of the experiment. PMID- 12744159 TI - [Temperature control in a combined therapeutic system for tumor]. AB - Tumor combined therapeutic system, a kind of medical instrument to restrain and kill tumor, is based on radio frequency heating technique and biological heating effect theory. It can be used solely or in combination with chemotherapy and actinotherapy for tumor hyperthermia. Combined with hardware measurement and adjustment, as well as application of the improved PID algorithm to the system of temperature control, the required accurate temperature control which directly influences the curative effect is realized. The experimental results presented in this paper demonstrated that the equipment works stably and reliably, and it is of good usefulness. PMID- 12744160 TI - [A project of magnetic coils newly designed to restrain the negative value of the intensity of magnetic induced electric field]. AB - In the non-invasive magnetic nerve stimulation of human brain, the coil of figure "8" is mostly used. The peak maximum is corresponding to the center of the coil, and the negative peak value is corresponding to the twain edges of the coil, the latter one is apt to cause abnormal nerve excitation. In this text, we put emphasis on the study of abnormal stimulation induced in magnetic stimulation and analyse the electric field intensity of the figure-8 coil. Aiming at the large negative value of intensity, we develop a new structure coil to restrain abnormal stimulation. Theoretical simulation is demonstrated here also. PMID- 12744161 TI - [In vivo measurement of rabbits brain impedance frequency response and the elementary imaging of EIT]. AB - The in vivo measurements of rabbit brain tissue impedance were taken under both normal and ischemic conditions by using two-electrode measurement method in the frequency range from 0.1 Hz to 1 MHz. The dynamic images about the resistivity of cerebral ischemia were reconstructed based on a 16-electrode system. The results of in vivo measurement showed that the ratio of impedance increased can be as high as 75% at frequencies lower than 10 Hz. In the range from 1 KHz to 1 MHz, the ratio showed a constant value of 15%. The electrical impedance tomography (EIT) images obtained suggested that the regions of impedance changes highly correspond to the position of ischemia. It is confirmed that the brain function changes caused by local deficiency of blood can be detected and imaged by EIT method. PMID- 12744162 TI - [On the correlation between the texture of contrast agent ultrasonic image and the intracardiac pressure]. AB - Based on the application of box-counting fractal model to the texture analysis of ultrasonic image after intravenous injection of Levovist acoustics contrast agent in a dog, this paper presents a method to calculate the fractal dimension(D) and assistant characteristic (C(L)) of the ultrasonic images of left ventricle. It was found that the D and C(L) changed regularly in the continued cardiac cycles, that is, the maximum value of D appears during diastasis, the minimum value appears during end-systole; as the ventricular systole begins, D changes from maximum to smallness, and as the diastole begins, D changes from smallness to maximum; the maximum value of C(L) appears around end-systole, the minimum value appears around diastasis; from the start of left ventricular systole to end systole, C(l) shows a tendency to change from smallness to maximum, and from the start of diastole to diastasis, it shows a tendency to change from maximum to smallness. The changes of D and C(L) are very similar to the changes of left ventricular pressure. It is thus evident that there is correlation between the fractal texture characteristics of the contrast agent ultrasonic images and the changes of intracardiac pressure. The results demonstrate that it is possible to measure the intracardiac pressure noninvasively. PMID- 12744163 TI - [The preprocessing of subtraction and the enhancement for biomedical image of retinal blood vessels]. AB - Image segmentation is still a difficult problem since its effect would vary with the subjects processed. An approach of subtracting background from the entire image of retinal blood vessels presented in this paper. The background subtraction is based on the real image itself taken photographically and is not dependent on the prior knowledge of system for recording image, the approach achieves the grayscale enhancement of retinal blood vessels in preprocessing and provides a quality image for the next process of binarization. This experiment in the preprocessing of subtraction shows good enhancement effect. PMID- 12744164 TI - [Application of complexity sequence in sleep staging based on sleep EEG data]. AB - In this paper an approach of time-window complexity sequence is applied to sleep EEG analysis. This approach can reduce the loss of state information due to the nonstationarity of EEG signal and the unevenness of state space, and can overcome certain limitations of the complexity itself to some extent. It will help to extract the state features of EEG in different sleep stages. In addition, we preprocess EEG by adopting ICA and wavelet transform (WT). The results show that some physiological artifact in EEG can be eliminated effectively by these methods, and the sleep staging based on sleep EEG data will be more exact. PMID- 12744165 TI - [Recombinant human brain myelin basic protein and its antibody]. AB - We constructed the expression vector by inserting 21.5 KDa MBP human brain full length cDNA coding sequence digested with restriction enzyme EcoR I and Sal I into downstream of pGEX-5T expression vector. The recombinant vector p5TMP was transformed into E. coli and the positive clonies were selected and incubated in LB medium induced by IPTG (isopropyl- -D-thiogalactoside). A new polypeptide band with apparent molecular weight 42 KDa was detected in transformed cell lysates by SDS-PAGE. Western blotting analysis confirmed that this fusion protein reacted specifically with antibodies to MBP, the expression level of MBP was about 414.6 mg/L medium estimated by immuno-dot blot, ELISA and absorbance scanning. Newzealand rabbits were immunized by subcutaneous injection of the purified recombinant MBP. The titer was obtained at 1:16 after 5 injections. The specificity of the antibody to MBP was confirmed by immuno-blot and Western blotting. PMID- 12744167 TI - [Synthesis of a triple helix-forming phosphorothioate oligodeoxynucleotides and its effects on coagulation activity of tissue factor (TF) and TF gene expression in endothelial cells]. AB - This study sought to synthesize a triple helix-forming phosphorothioate oligodeoxynucleotides (TFO-ps) and assess its effects on coagulation activity of the tissue factor(TF) and TF gene expression in endothelial cells. Experiment antiparallel oligodeoxynucleotides T21GTa sequence was designed and synthesized by phosphoramidite method and decorated with all-PS linkage. The affinity of TFO and TFO-ps was determined by electrophoretic mobility shift assays(EMSA). Cellular uptake of [32]P-labeled TFO-ps and the effect of TFO-ps on TF gene expression and coagulation activity of TF were measured in endothelial cell strain ECV304. The results showed that TFO-ps (T21GTa-ps) formed a triplex binding in antiparallel orientation to the puring-rich target strand-SSRE, with Kd value of 3.6 x 10(-10) M. The uptake rate of TFO-ps (T21GTa-ps) in ECV304 was about 11.65%. This compound mainly localized within the nucleus sediment(77.25%), significantly reduced the average OD of the mRNA expression and protein synthesis of TF gene, and obviously decreased the coagulation activity of TF. In conclusion, TFO-ps (T21GTa-ps) shows obvious anti-coagulation activity and its mechanism involves the inhibition of TF gene expression. PMID- 12744168 TI - [Study on the expression of telomerase RNA in leukocyte]. AB - Total cDNA of human telomerase RNA(hTR) gene was cloned by means of RT-PCR and inverted into retroviral vector (pLNCX) to construct the mammalian cell expression plasmid. Then, by using lipofectin-mediated DNA transfection, the obtained expression plasmid was successfully transfected into human normal peripheral blood leukocyte. All data suggested that expression of transfected exogenous hTR gene can not reconstitute telomerase activity. Flow cytometry analysis and data from cell growth curve also indicated that expression of exogenous gene can not prolong the longevity of leukocyte, but rather inhibit the growth of leukocyte and induce its apoptosis. We conclude that expression of exogenous gene may block the coalition of telomerase RNA and its catalytic subunit(hTRT) and block the coalition of telomerase RNA template and telomere DNA, thus affecting telomerase activity and repressing cell proliferation. PMID- 12744166 TI - [The roles of VEGF and C-myc in occurrence, development and metastasis of gallbladder carcinoma]. AB - To investigate the relationship of the expression of vascular endothelial growth factor (VEGF) and C-myc with the occurrence, development and metastasis of gallbladder carcinoma, the expression levels of VEGF and C-myc in gallbladder carcinoma tissue (n = 30) and in normal gallbladder tissue (n = 20) were examined by immunochemistry. Results show that the positive rates of VEGF and C-myc in gallbladder carcinoma tissue were 80% and 63.3% respectively, and 45% and 25% respectively in normal gallbladder tissue. The positive rates of VEGF and C-myc were significantly higher in gallbladder carcinoma than in normal gallbladder tissue. The expression of VEGF and C-myc in gallbladder carcinoma related to the metastasis of gallbladder carcinoma. VEGF and C-myc play important roles in the occurrence, development and metastasis of gallbladder carcinoma. PMID- 12744169 TI - [A logistic cellular automaton for simulating tumor growth]. AB - This paper focuses on a differential equation logistic model simulating tumor growth. We design a kind of tumor dynamic growth model with one-dimensional cellular automata. A discrete logistic model is developed from the continuous logistic model. Based on others' work, we design discrete mathematical growth dynamic model with cellular automaton. In terms of discrete model, we design stochastic evolving rules of cellular automaton. And this paper simulates the tumor growth dynamic model with cellular automata. The theoretic analysis and results of cellular automaton model are in agreement with data from the ideal differential equation logistic growth of cancer. PMID- 12744170 TI - [Cardiomyocytes membrane channel currents and their dynamics]. AB - The mathematical models for simulation of cardiac sodium, potassium and calcium channel kinetics courses and currents were developed to simulate the properties of ionic currents and channel dynamic courses. With modifications of these models, it is possible to make them integrated for simulating the whole process of action potential, thus additional discussion on ionic mechanism could provide a theoretical foundation for further animal experiments and clinical applications. PMID- 12744171 TI - [Simulation application of whole-heart electophysiological model]. AB - We invented an efficiently approach to building whole-heart electrophysiological model with cellular automata style massive parallel computing. In this paper, we introduce the simulation facilities of a model we built and the potential application of such model. The first use is to solve the forward problem of electrocardiogram. Simulating arrhythmia ECG and accurately interpreting the significance of various irregular waveforms will be the key goal. Investigating the dynamic property of cardioelectrical activity at cellular and channel levels is the second application, aiming at revealing the mechanism of the generation and sustentation of arrhythmia. Third, the model can be used to research the impacts of artificial interventions on cardioelectrical activity. Electrical defibrillation and pace-making as well as the use of channel block agents are all cases. PMID- 12744173 TI - [Study of 125I-interleukin-8 distribution in mice]. AB - IL-8 was radiolabeled with 125I via Bolten-Hunter agent and the distribution of 125I-IL-8 in mice was determined in order to understand IL-8 behaviors in vivo. The percentage of 125I-IL-8 in blood, heart, liver, lung, kidneys, bone and spleen was obtained. The half-clearance of fast phase T1/2 alpha and slow phase T1/2 beta were 0.32 h and 8.01 h respectively. Most of 125I-IL-8 was excluded by kidneys. PMID- 12744172 TI - [Effect of shenfu injection on microcirculation]. AB - This study was aimed to assess the effect of Shenfu injection on different circulation state. Using a microcirculation microscope system, we observed mice's auricle micro-artery diameter, density of capillary, blood velocity in different circulation state (i.e. normal state, epinephrine or endotoxin induced microcirculation disturbance state) after administering Shenfu injection into their caudal vein, and we compared the Shenfu group with Shenmai group and Dexamethasone group. The results showed that Shenfu injection causes the auricle microartery diameter to enlarge and the density of capillary and blood velocity to increase in different microcirculation state, and such effect is especially notable on the epinephrine induced microcirculation disturbance group and endotoxin induced microcirculation disturbance group; the effect of Shenfu injection is stronger than that of Shenmai injection and similar to Dexamethasone injection. In addition, Shenfu injection was shown to have remarkable effect on resisting the lowering of limb temperature when the mice are attacked by endotoxin. PMID- 12744174 TI - [The effect of magnitopuncture stimulation on HRV during simulated driving under vibration conditions]. AB - The purpose of this study was to assess the effect of magnitopuncture stimuli for reducing driver mental stress and fatigue using power spectral analysis of the heart rate variability (HRV) and subjective evaluation. The experiments were divided into A-group and B-group. In both groups the subjects performed the simulator for 90 minutes under a vibration conditions with an erect sitting posture in a silent environment, and magnitopuncture was put on the acupoints when performing the task for one hour in A-group. In this study HRV exhibited a significant difference between the two groups after the simulating task (P < 0.05). A conclusion that magnitopuncture stimuli can reduce the driver mental stress and fatigue effectively was drawn. PMID- 12744176 TI - [Component distribution in gradient biomaterial prepared with multi grades energy PIII]. AB - During the preparation of functional gradient materials (FGM) with plasma immersion ion implantation-ion beam enhanced deposition (PIII-IBED), the combination strength between the coating and the substrate would be greatly affected by the implantation dose and the distribution of implanted ions in substrate. According to the requirements of FGM, an idea of multi grades energy implantation had been suggested, with which the Gauss peak could be moved toward the surface, and the concentration of implanted ions could be maximized at the surface. In this study, the distribution of carbon ions implanted into titanic alloy substrate have been simulated theoretically, and the tentative idea have been confirmed experimentally. PMID- 12744175 TI - [Effect of temperature on the heart function of the five-strip eyebrow squirrel]. AB - This is an in vitro study designed to assess the effect of temperature variation on the heart function of the five-strip eyebrow squirrel--amammal of the hibernating kind. A heart model of exsomatizing action and for determining the physiological parameters with four physiological recorders was used to measure the LVSP, LVEDP and (+/- dP/dtmax), respectively. It was found that the curves of the squirrel's heart mechanics property in relation to temperature variation are similar to the curves of the non-hibernating animals'. But, the squirrels' heart contraction function changes with in a much wider range, suggesting that their hearts are more tolerant of temperature variation. PMID- 12744177 TI - [Image reconstruction in electrical impedance tomography based on genetic algorithm]. AB - Image reconstruction in electrical impedance tomography (EIT) is a highly ill posed, non-linear inverse problem. The modified Newton-Raphson (MNR) iteration algorithm is deduced from the strictest theoretic analysis. It is an optimization algorithm based on minimizing the object function. The MNR algorithm with regularization technique is usually not stable, due to the serious image reconstruction model error and measurement noise. So the reconstruction precision is not high when used in static EIT. A new static image reconstruction method for EIT based on genetic algorithm (GA-EIT) is proposed in this paper. The experimental results indicate that the performance (including stability, the precision and space resolution in reconstructing the static EIT image) of the GA EIT algorithm is better than that of the MNR algorithm. PMID- 12744178 TI - [Design and realization of HL7 gateway]. AB - With the development of the technology in medical informatics area, Health Level Seven (HL7) Standard for electronic data exchange in all healthcare environments has become a widely used standard. To bring it into practice in China, we developed an HL7 gateway. The gateway is used to connect two different medical information systems, a Hospital Information System (HIS) and a Laboratory Information Management System (LIMS), via the HL7 message exchanging. The system verified the feasibility of HL7 standard and made a good example for further research and development. PMID- 12744179 TI - [The effect of high pressure on morphology, expression of alpha-actin and proliferating cell nuclear antigen of vascular smooth muscle cells--a new model of common carotid artery exposed to stress in vivo]. AB - To investigate the role and mechanism of mechanical stress on arterial remodeling, a new model of common carotid artery exposed to stress in vivo was established in rat, in which the change of pressure is the only influencing factor. The effect of high pressure on the morphology and expression of alpha actin and proliferating cell nuclear antigen (PCNA) of vascular smooth muscle cells (VSMCs) in the artery was assessed. The arteries were perfused by both high pressure (160 mmHg) and normal pressure (80 mmHg) for 6 hours. The changes of morphology, expression of alpha-actin and PCNA in VSMCs were studied by histology and immunohistochemistry. The results showed that the new model could be controlled well in pressure and frequency. The euchromatin was increased and PCNA positive particles were observed in the nuclei of VSMCs, but the expression of alpha-actin was decreased when the arteries were exposed to the high pressure. The new model has been successfully established, which provides a new tool for studying the effect of mechanical stress on arterial remodeling. In this experiment, VSMCs underwent a transformation from contractile phenotype into synthetic phenotype and tended to proliferate in response to the high pressure. PMID- 12744181 TI - [The research of noninvasive cardiovascular diagnosis and test instrument]. AB - The study of pulse wave, including its quantitative analysis and propagation, is one of the hot topics in the fields of clinical medicine, such as how to detect cardiovascular diseases and make early diagnosis without harm. In this paper are discussed a new type of automatic cardiovascular diagnosis method and the corresponding test instrument. The cardiovascular system parameters can be achieved through the wave changes, related data processing and analyzing. This system includes piezo-film transducer, signal amplifier, data collecting and data analyzing equipment. Various pulse diagrams can be shown in omnidirectional and multifunctional ways. The measurement error can be reduced effectively by introducing Mini-wave analysis in the design of software. The research indicates that combination of this equipment with the traditional Chinese medicine will provide wide practical application. PMID- 12744180 TI - [Research and development on the dynamic system for detecting the force of acupuncture needle during the acupuncture process in the clinical practice of traditional Chinese medicine]. AB - Manipulations are essential to the acupuncture curative effect. With the technique limited, the researches on acupuncture manipulations can only be carried out with respect to the senses and experience of doctors and patients, thus seriously impeding the modernization and internationalization of acupuncture. Both the modern integration technology of transducer and the biomechanical principles are applied to develop a detection system that can measure the interaction force between the manipulator and the manipulated patient on the needle at each kind of manipulation. Through clinical practice, the force waveforms acting on needle with the manipulations of symmetrical twirling rotating and symmetrical lifting-inserting were recorded so as to realize the quantitative, objective and real-time detection of the force during the acupuncture process, which provided a new experimentation means and analysis method for the improvement of clinical curative effect and quantitative research of acupuncture and meridians. PMID- 12744183 TI - [Refit of simple plasma generator]. AB - A simple plasma generator was refitted on the basis of usual ion sputter in biological labs, and uniformity of plasma intensity was verified. Bacteriological dishes were used as an example of material surface modification. The results showed that bacteriological dishes were switched from hydrophobic to hydrophilic after plasma treatment, and they obtained the ability to support cells adhesion and spreading. This demonstrated that the refitted ion sputter can be used as an effective plasma generator for biomaterial surface engineering. PMID- 12744184 TI - [The synthesis of polyanhydrides and its application in biomedical field]. AB - This paper reviews the development of the polyanhydrides as a new biodegradable polymer, and highlights the methodological and technological progress in the synthesis of the polymer. Subsequently, the future researches and developments of polyanhydrides are prospected. PMID- 12744182 TI - [The development of a pocket blood pressure holter based on handhold personal computer]. AB - This paper introduces a kind of pocket blood pressure holter based on handhold personal computer. The measurement of blood pressure is controlled by ultralow power consumption MCU. The system configures interface circuit with handhold personal computer. The BP data can be stored, displayed, analysed and transmitted by handhold personal computer. The measurement parameters can be modified. The hardware is made up of three units: simulation circuit unit, intelligent control unit and interface circuit unit. Every unit structure and its principle are presented. The software of obtaining the BP data and communication with handhold personal computer are also illuminated. The system possesses the characters of portability, intelligence, convenience, high precision, ultralow power consumption and stable capability. It is possible to become an ideal blood pressure monitor in family. PMID- 12744185 TI - [The related research of tissue-engineered tendon in vivo]. AB - Using tissue-engineered tendons to repair tendons and ligaments as well as functional reconstruction is the focus of nowadays researches. The scaffolds must be not only unharmful to health, but also easy for cells attachment, and be able to induce collagen deposition to form a neotendon with mechanic properties similar to those of normal tendon. In recent researches, it has been found that the mechanic properties of the implants change with the degrading and femdonizing of scaffolds. The relationships between collagen deposition, scaffolds degradation and mechanic properties of neotendon need to be defined more clearly. PMID- 12744186 TI - [Researches on PEG-modified copolymer nanoparticle]. AB - Biodegradable polymeric nanoparticles acting as drug carrier have important potential applications such as site-specific drug delivery and controllable drug delivery. However, these carriers cannot generally be used because they are eliminated by the reticulo-endothelial system within seconds or minutes after intravenous injection. To overcome this limitation, more and more researchers introduce hydrophilic polyethylene glyeol(PEG) to modify polymeric nanoparticles for avoiding their uptake by reticulo-endothelial system. Introducing PEG not only changes polymer nanoparticles' biodegradation in vivo, but also influences drug's properties such as drug release, in vivo biodistribution, et. al. In this paper are reviewed the researches of PEG-modified copolymer nanoparticles, including their preparation and size distribution, stability, drug incorporation, drug release, in vivo biodistribution, in vitro cytotoxicty. A prospect for the researches and developments of the PEG-modified copolymer nanoparticles was also made. PMID- 12744187 TI - [Bone tissue engineering scaffolds]. AB - Bone tissue engineering may provide an alternative to the repairs to skeletal defects resulting from disease, trauma or surgery. Scaffold has played an important role in bone tissue engineering, which functions as the architecture for bone in growth. In this paper, the authors gave a brief introduction about the requirement of bone tissue engineering scaffold, the key of the design of scaffolds and the current research on this subject. PMID- 12744188 TI - [Progress in scaffolding materials of bioartificial liver]. AB - The key materials for bioartificial liver (BAL) construction include biomaterials and scaffolding materials. The former mainly refers to hepatocytes, nonparenchymal cells, etc. The latter mainly refers to films and other scaffolding materials, the properties of which correlate directly with hepatocyte growth and functions, and thus are related to the support effects of BAL. Several kinds of scaffolding materials frequently used for BAL construction in recent years are reviewed in this article. PMID- 12744189 TI - [Application of weighted minimum-norm estimation with Tikhonov regularization for neuromagnetic source imaging]. AB - In magnetoencepholography(MEG) inverse research, according to the point source model and distributed source model, the neuromagnetic source reconstruction methods are classified as parametric current dipole localization and nonparametric source imaging (or current density reconstruction). MEG source imaging technique can be formulated as an inherent ill-posed and highly underdetermined linear inverse problem. In order to yield a robust and plausible neural current distribution image, various approaches have been proposed. Among those, the weighted minimum-norm estimation with Tikhonov regularization is a popular technique. The authors present a relatively overall theoretical framework Followed by a discussion of the development, several regularized minimum-norm algorithms have been described in detail, including the depth normalization, low resolution electromagnetic tomography(LORETA), focal underdetermined system solver(FOCUSS), selective minimum-norm(SMN). In addition, some other imaging methods, e.g., maximum entropy method(MEM), the method incorporating other brain functional information such as fMRI data and maximum a posteriori(MAP) method using Markov random field model, are explained as well. From the generalized point of view based on minimum-norm estimation with Tikhonov regularization, all these algorithms are aiming to resolve the tradeoff between fidelity to the measured data and the constraints assumptions about the neural source configuration such as anatomical and physiological information. In conclusion, almost all the source imaging approaches can be consistent with the regularized minimum-norm estimation to some extent. PMID- 12744190 TI - [Review of evaluation techniques for medical image reconstruction]. AB - This article reviews the research progress in qualitative evaluation of medical image reconstruction, especially the binary image reconstruction. According to the affected point and evaluation character, the evaluation criteria are classified into four categories. Based on the brief introduction of all kinds of evaluation criteria, the paper makes an analytical comparison, and points out their merit and fault. This study provides a useful basis for selection and design of medical image reconstruction algorithms. PMID- 12744191 TI - [Application of image analysis system to the study of biomedicine]. AB - Image analysis plays an important role in morphological study of tissues and cells and their quantitative analysis. It also contributes to clinical pathological diagnosis. With the rapid development of computer technology great progress has been made in the image analysis system, its measurement, rapidity, accuracy and the extent of automation have been greatly enhanced. In the meantime advances in medicine give impetus to its improvement. In this paper, the process of development, the basic structure and mechanism of image analysis system are discussed and the application of image analysis system to the study of biomedicinal is presented. PMID- 12744192 TI - [Analysis of DICOM and its application in teleradiology]. AB - Digital Imaging Communications in Medicine(DICOM) was brought forward by ACR and NEMA to solve the joining problem of medical equipment. It is a public international standard now. Using DICOM in teleradiology is very important, especially for China. In this paper the authors analyze the contents and specialty of DICOM and make comments on how to use it in teleradiology and picture access and communication system(PACS). PMID- 12744193 TI - [Mechanism of priming cytotoxic T cell response and strategy for enhancing DNA vaccine potency in DNA immunization]. AB - DNA vaccination that can induce both cellular and humoral immune response has become an attractive immunization strategy against cancer and infectious disease. Elucidation of the precise mechanisms of immune priming will be important in the development of effective DNA vaccines. In this review, we illustrate possible mechanisms in priming cytotoxic T cell response involving the intracellular degradation, processing and presentation of encoded antigen. We also discuss the roles of costimulatory molecules expressed on antigen-presenting cells (APCs) in inducing optimal CTL activity. Hence, a rational strategy for increasing DNA potency would be to facilitate these pathways. Additionally, we focus on recent strategies including rapid degradation of ubiquitin-antigen fusion proteins, direct targeting to APCs for increased DNA uptake, direct routing an antigen into the MHC class I and II processing and presentation pathways, and increasing the immunogenicity of encoded antigen. All of these approaches have resulted in increased potency of DNA vaccines. PMID- 12744194 TI - [Fatigue damage and repair in bone]. AB - Bone is a load-bearing organ in human body. Fatigue damage occurs readily at the modest loads to which bone is subjected during its habitual physiological usage. Even bone fracture may occur during vigorous activity. The nature of fatigue damage is that in bone there are very fine microcracks which are smaller than typical microcracks, and may occur at the level of hydroxyapatite crystals. But bone can repair microdamage by bone remodeling. Osteocytes play an important role of signaling during bone remodeling. Some researchers attempted to describe the process of bone fatigue damage and repair by mathematic, mechanical models in order to understand it well and to apply it well in clinical practice. PMID- 12744196 TI - How human immunodeficiency virus voluntary testing can contribute to tuberculosis control. PMID- 12744197 TI - Smallpox vaccination and the patient with HIV/AIDS. PMID- 12744198 TI - Hope for turning of the tide. PMID- 12744195 TI - TB can be cured. PMID- 12744199 TI - Needed: committed and daring voices. PMID- 12744200 TI - HIV care in 2003: a viewpoint. PMID- 12744201 TI - Will AIDS finally teach us the meaning of sustainable human development (for all)? PMID- 12744202 TI - Introduction. Cancer cytogenetics: methods and protocols. PMID- 12744203 TI - Cytogenetic studies in hematologic malignancies: an overview. PMID- 12744204 TI - The myeloid disorders: background. AB - Myeloid disorders do not usually present quite so many technical challenges to the cytogeneticist as does ALL: the chromosomes are often of a better quality, and white blood cell counts are not usually so high, except in CGL. Unlike in the chronic lymphoid disorders, there is no need for mitogens to include cell division. However, apart from the Ph in CGL, the overall frequency of detected clones is not so high. This has the consequence that a large proportion of patients is denied the diagnostic and prognostic benefit of knowing the cytogenetic abnormalities that are associated with their disease. PMID- 12744205 TI - Cytogenetic techniques for myeloid disorders. PMID- 12744206 TI - Acute lymphoblastic leukemia: background. AB - Acute lymphoblastic leukemia is a fascinating disease for the cytogeneticist, as so many cases have a clone detectable by cytogenetics or FISH, and because identifying the abnormalities provides such useful information to the clinician. However, it is also a frustrating disease, as it has technical challenges such as a marked tendency for the sample to clot during harvesting, frequently poor chromosome morphology, and, especially in the high count cases, failure to provide any divisions at all for analysis. For these reasons, this book includes two chapters on the practical aspects of undertaking cytogenetic studies in ALL to illustrate contrasting approaches. The first is from a laboratory that is a world leader in its success rates, which has an enviably low sample/cytogeneticist ratio, and which is usually able to expect a good-sized sample commensurate with the importance given to a diagnostic cytogenetic study. The second is from a laboratory that also has a good success rate, despite having to cope with a higher workload and often much smaller samples. This is not to imply that each technique is limited to such circumstances; both are worthy of study and emulation. PMID- 12744207 TI - Conventional cytogenetic techniques in the diagnosis of childhood acute lymphoblastic leukemia. PMID- 12744208 TI - Chromosome preparations from bone marrow in acute lymphoblastic leukemia: cytogenetic techniques. PMID- 12744209 TI - Lymphoid disorders other than common acute lymphoblastic leukemia: background. AB - The cytogenetic abnormalities that are found in chronic lymphoid malignancies (and in acute leukemias deriving from relatively mature cells) fall mainly into two groups according to whether the malignant cells are of B-lineage or T lineage. In most of the B-lineage cases. there is some abnormality of the IgH gene which is located at 14q32 or, less frequently, the other immunoglobulin genes located at 2p12 and 22q11. In the T-lineage cases, there is often some abnormality involving the T-cell receptor loci, most frequently those at 14q11. Other abnormalities occur, but few have a close association with a particular disease type, and so do not often contribute to determining a precise diagnosis. This lack of specificity is due partly to deficiencies in our understanding of the biological relationships between different lymphoid disorders, and also to the various classifications that have been used. As a consequence, there has to be some doubt about the diagnosis assigned to many of the cases in published cytogenetic studies. It is also difficult to combine data from series that have used different classification systems. The present unsatisfactory situation greatly limits the clinical usefulness of cytogenetic studies and there is a real need to unravel the complexities of this large family of malignancies. PMID- 12744210 TI - Other lymphoid malignancies: cytogenetic techniques. PMID- 12744211 TI - Cytogenetic and genetic studies in solid tumors: background. AB - Cytogenetic and genetic studies of solid tumors are an area of continuing rapid growth and discovery. They provide an excellent resource for students interested in research, but more importantly have the potential to benefit patients. As has already happened in the leukemias, identifying certain genetic abnormalities can establish a definite diagnosis and/or can indicate likely response to treatment. Other chromosome abnormalities are ubiquitous, such as rearrangements of chromosome 1, and do not appear to correlate with other clinical features; however, even abnormalities of this sort serve to distinguish between malignancy and reactive conditions. Trisomy 7 is also common in many kinds of tumor and has been found as the sole abnormality in cells from tissues surrounding a tumor; the significance of this is not clear: Either trisomy 7 is associated with some reactive response in normal cells, or else it was a primary clonal abnormality and indicated the presence of infiltrating early malignant cells. PMID- 12744212 TI - Human solid tumors: cytogenetic techniques. PMID- 12744213 TI - Analysis and interpretation of cytogenetic findings in malignancy. PMID- 12744214 TI - Cytogenetic studies using FISH: background. PMID- 12744215 TI - FISH techniques. PMID- 12744216 TI - FISH, CGH, and SKY in the diagnosis of childhood acute lymphoblastic leukemia. PMID- 12744217 TI - Solving problems in multiplex FISH. PMID- 12744218 TI - Some difficult choices in cytogenetics. AB - In making a selection of features of these technologies, it is inevitable that some will be omitted that other cytogeneticists feel should have been included. The author could probably justifiably be accused of bias. However, based on experience in a laboratory that has used almost every type of assay mentioned in this chapter, the following opinions are offered about their current value in providing a routine malignancy cytogenetics service: 1. The foundation is still a conventional cytogenetic study, preferably with the use of an automated karyotyping system. 2. Added to this, there should be the capability of performing FISH studies using chromosome paints and gene-specific probes. Cytogenetics and FISH form a powerful partnership when backed by experienced cytogeneticists. MFISH or SKY are also useful if the laboratory can afford the considerable extra expense. CGH and fibre FISH are generally better suited to research projects, and at present have few applications in a routine diagnostics service. 3. At present, molecular methods such as RT-PCR mostly tend to produce results that have a greater need of confirmation by other techniques before they can be used for clinical management. PMID- 12744219 TI - Introduction to the analysis of the human G-banded karyotype. PMID- 12744221 TI - "I'm just a girl who can't say no"?: women, consent, and sex research. AB - Examinations of participant characteristics suggest that particular people volunteer to take part in social science research, and women have been found to be more likely to volunteer than men. However, within sex research it seems that this cooperation diminishes, and women are more likely to refuse to participate (particularly if a study seems personal or invasive). Evidently, this affects the reliability and validity of research and could lead to women's sexual health care decisions based on unrepresentative sampling. Given the need to include women in sex research, this article examines reasons for refusal and suggests improvements in research methods. PMID- 12744220 TI - Neuroticism does not affect cognitive functioning in later life. AB - In a cross-sectional study, Jorm and colleagues (1993. Personality and Individual Differences, 15, 721-723) found that neuroticism was related to poorer cognitive performance in the elderly. The present study was initiated to expand their findings using both cross-sectional and longitudinal data from the Maastricht Aging Study. In contrast to the findings of Jorm and coworkers, neuroticism was not associated with either current cognitive performance or cognitive decline over a period of 3 years. PMID- 12744222 TI - Current status of endothelin blockade for the treatment of cardiovascular and pulmonary vascular disease. AB - With recognition of the significant role of the endothelin system in cardiovascular and pulmonary vascular diseases, attention has turned to the therapeutic application of agents that antagonize this system. Three strategies can be employed: dual endothelin receptor (ETA and ETB) antagonism, selective ETA antagonism and inhibition of endothelin-converting enzymes. The first two strategies have been evaluated in late-phase clinical trials, with mixed results. The use of the dual endothelin receptor antagonist bosentan in pulmonary arterial hypertension has been successful. Available data are less compelling, but nonetheless promising, for the use of bosentan in digital ulceration secondary to systemic sclerosis, and tezosentan (another dual receptor blocker) in acute heart failure. Both types of receptor antagonists, however, have been evaluated in systemic hypertension and chronic systolic heart failure and are unlikely to be further developed in these areas. For the treatment of hypertension, their (moderate) efficacy and safety/tolerability profiles appear no more favorable than existing antihypertensive agents. In terms of treatment for chronic heart failure, these agents appear to offer no further benefit over standard therapy for the treatment of chronic heart failure, and in fact, may be associated with poorer outcomes. The complexity of the endothelin system and its diverse roles in multiple disease states will ensure its position as a target for drug development. PMID- 12744223 TI - Eplerenone. Pharmacia. AB - Pharmacia Corp (formerly GD Searle & Co) is developing eplerenone, an aldosterone receptor antagonist, as a potential treatment for congestive heart failure and systemic hypertension. The compound has been registered for hypertension and Pharmacia plans to submit a supplemental NDA for congestive heart failure in the first half of 2003 based on positive results from a phase III clinical trial. PMID- 12744224 TI - Molecular cloning of alpha-amylases from cotton boll weevil, Anthonomus grandis and structural relations to plant inhibitors: an approach to insect resistance. AB - Anthonomus grandis, the cotton boll weevil, causes severe cotton crop losses in North and South America. Here we demonstrate the presence of starch in the cotton pollen grains and young ovules that are the main A. grandis food source. We further demonstrate the presence of alpha-amylase activity, an essential enzyme of carbohydrate metabolism for many crop pests, in A. grandis midgut. Two alpha amylase cDNAs from A. grandis larvae were isolated using RT-PCR followed by 5' and 3' RACE techniques. These encode proteins with predicted molecular masses of 50.8 and 52.7kDa, respectively, which share 58% amino acid identity. Expression of both genes is induced upon feeding and concentrated in the midgut of adult insects. Several alpha-amylase inhibitors from plants were assayed against A. grandis alpha-amylases but, unexpectedly, only the BIII inhibitor from rye kernels proved highly effective, with inhibitors generally active against other insect amylases lacking effect. Structural modeling of Amylag1 and Amylag2 showed that different factors seem to be responsible for the lack of effect of 0.19 and alpha-AI1 inhibitors on A. grandis alpha-amylase activity. This work suggests that genetic engineering of cotton to express alpha-amylase inhibitors may offer a novel route to A. grandis resistance. PMID- 12744225 TI - [2002 Standards, Options and Recommendations for the use of [18F]-FDG (PET-FDG) in cancerology]. PMID- 12744226 TI - Hemostatic variables, carbohydrate metabolism and lipid profile in women with low body mass index. AB - The aim of this study was to evaluate hemostatic variables in women according to different body mass index (BMI) values, and then correlate them with some metabolic parameters - fasting insulin and glucose, total cholesterol, high density lipoprotein (HDL)-cholesterol, low-density lipoprotein (LDL)-cholesterol and triglycerides. Eighty-four female patients aged 18-39 years were recruited, and agreed to participate in the study. The study group was divided into three subgroups according to BMI: low BMI (BMI < 18.5 kg/m2; n = 43), normal-weight (control) (BMI 18.5-24.99 kg/m2; n = 21) and overweight/obese (BMI > 25 kg/m2; n = 20). BMI was calculated, and the following measurements were taken: International Normalized Ratio, antithrombin III, tissue plasminogen activator (t PA) activity, t-PA-antigen, plasma fibrinogen level, factor VII, Plasminogen activator inhibitor (PAI)-1 activity and antigen and metabolic parameters: fasting insulin and glucose, total cholesterol, HDL-cholesterol, LDL-cholesterol and triglycerides. The results were statistically analyzed. In the low BMI group, a negative correlation between fasting insulin and PAI-1 activity, and a positive correlation between fasting glucose and PAI-1 antigen were observed. Also, a strong negative correlation between PAI-1 activity and insulin/glucose index was found. Plasma insulin levels were significantly lower in the low-BMI women than in the overweight/obese group (p < 0.001) and with no difference compared to the control group. We did not find any difference in fasting glucose level between all groups. HDL-cholesterol showed the highest levels in the normal BMI group and was significantly higher than in the low BMI and obese groups (p < 0.05 and p < 0.01, respectively). PAI-1 activity in the low BMI women revealed increased activity in comparison to control and overweight/obese women (p < 0.001 and p < 0.05, respectively). Lower antigen levels were also shown as compared to both these groups (p < 0.001 and p < 0.001, respectively). Similar results were obtained with t-PA antigen levels (p < 0.001 and p < 0.001, respectively). There were no differences in activity of t-PA in all groups. Obese women showed significantly higher fibrinogen levels than other BMI groups (p < 0.001 and p < 0.001 respectively). Analysis of hemostatic variables in women with a low BMI testify to the impaired fibrinolysis in this group, also showing a strong correlation with carbohydrate metabolism. PMID- 12744227 TI - Of cells, mice, men and radiobiology. PMID- 12744228 TI - Radiolabelled white blood cells and inflammatory bowel disease. PMID- 12744229 TI - Guidelines for the provision of radiopharmacy support to nuclear medicine. PMID- 12744230 TI - Truth, science, and advertising in the information age. PMID- 12744231 TI - Special issue dedicated to Dr. Stanislav Tucek. PMID- 12744232 TI - Abstracts of the 5th International Symposium on Leukemia and Lymphoma. 12-15 March 2003. Amsterdam, The Netherlands. PMID- 12744233 TI - [Proceedings and abstracts of the National Week of Gastroenterology. Queretaro, Mexico, 23-24 November 2002]. PMID- 12744234 TI - Proceedings of the 5th International Symposium on the Biological Oxidation of Nitrogen in Organic Molecules. Munich, Germany, October 4-6, 2001. PMID- 12744235 TI - Bookmark this site. A guide to home care & hospice websites. AB - An information source like no other, the Internet is a place for communication, research, community building, and information sharing. As the Internet has grown in popularity, the number of sites providing useful information and networking has grown exponentially. The challenge to time strapped home care and hospice providers is finding those sites that provide the information you need to know and getting to that information quickly and easily. PMID- 12744236 TI - Discussion on frequency of the HIV-protective CC chemokine receptor 5-Delta 32/Delta 32 genotype is increased in hepatitis C. PMID- 12744237 TI - Frequency of the 32-base pair deletion in the chemokine receptor CCR5 gene is not increased in hepatitis C patients. PMID- 12744238 TI - Discussion of the role of hemochromatosis susceptibility gene mutation in protecting against iron deficiency in celiac disease. PMID- 12744239 TI - Abstracts of the Chemotherapy Foundation Symposium XX: Innovative Cancer Therapy for Tomorrow. November 13-16, 2002, New York, New York, USA. PMID- 12744240 TI - Discussion of the role of hemochromatosis susceptibility gene mutation in protecting against iron deficiency in celiac disease. PMID- 12744241 TI - Discussion on the effect of chronic hypergastrinemia on human enterochromaffin like cells: insights from patients with sporadic gastrinomas. PMID- 12744242 TI - Survivin in liver. PMID- 12744244 TI - [Abstracts of the LVII Congress of the Brazilian Society of Cardiology. Sao Paulo, Brazil, 2002]. PMID- 12744243 TI - [Abstracts of the 109th Congress of the French Society of Ophthalmology. 10-14 May 2003, Paris, France]. PMID- 12744245 TI - Digestive Disease Week. Abstracts of the American Society for Gastrointestinal Endoscopy meeting. 18-21 May 2003, Orlando, Florida, USA. PMID- 12744246 TI - Analgesia for acute myocardial infarction pain. PMID- 12744247 TI - Emergency physician rights and self-disclosure. PMID- 12744249 TI - 2003 Membership list of the Academy of Medicine of Belgium. PMID- 12744248 TI - Use of ultrasonography in the emergency department: time for a reappraisal. PMID- 12744250 TI - Validity of D-dimer tests for pulmonary embolism: better to diagnose or exclude? PMID- 12744251 TI - Emergency department-based atrial fibrillation treatment strategy with low molecular-weight heparin. PMID- 12744252 TI - Typical chest pain and onset of acute myocardial infarction in a prospective cohort of emergency calls. PMID- 12744253 TI - Blood lactate as a prognostic marker in acetaminophen-induced acute liver failure. PMID- 12744254 TI - [Abstracts of the 15th Spring Meeting of the Japanese Society of Allergology. Yokahama, Japan, May 12-14, 2003]. PMID- 12744255 TI - Conflict resolution in emergency medicine. PMID- 12744257 TI - Abstracts of the 76th Annual Meeting of The Japanese Pharmacological Society. March 24-26, 2003. Fukuoka, Japan. PMID- 12744256 TI - Proceedings of the 2003 Spring Meeting of the WPSA UK Branch. PMID- 12744258 TI - Abstracts of the annual main meeting of the Society for Experimental Biology. 31 March-4 April 2003, Southampton, United Kingdom. PMID- 12744259 TI - Abstracts of the 69th Annual meeting of the German Society of Cardiology--Heart and Circulatory Research. 24-26 April 2003, Mannheim, Germany. PMID- 12744260 TI - Displays in space. AB - This chapter describes the human and environmental factors that dictate the way that displays must be designed for, and used in space. A brief history of the evolution of such display systems covers developments from the Mercury rockets to the International Space Station. PMID- 12744261 TI - A comparative bear model for immobility-induced osteopenia. AB - The National Institutes of Health (NIH) and the National Aeronautics and Space Administration (NASA) are seeking solutions to the human problem of osteopenia, or immobility-induced bone loss. Bears, during winter dormancy, appear uniquely exempted from the debilitating effects of immobility osteopenia. NIH and ESA, Inc. are creating a large database of metabolic information on human ambulatory and bedrest plasma samples for comparison with metabolic data obtained from bear plasma samples collected in different seasons. The database generated from NASA's HR113 human bedrest study showed a clear difference between plasma samples of ambulatory and immobile subjects through cluster analysis using compounds determined by high performance liquid chromatography with coulometric electrochemical array detection (HPLC-EC). We collected plasma samples from black bears (Ursus americanus) across 4 seasons and from 3 areas and subjected them to similar analysis, with particular attention to compounds that changed significantly in the NASA human study. We found seasonal differences in 28 known compounds and 33 unknown compounds. A final database contained 40 known and 120 unknown peaks that were reliably assayed in all bear and human samples; these were the primary data set for interspecies comparison. Six unidentified compounds changed significantly but differentially in wintering bears and immobile humans. The data are discussed in light of current theories regarding dormancy, starvation, and anabolic metabolism. Work is in progress by ESA Laboratories on a larger database to confirm these findings prior to a chemical isolation and identification effort. This research could lead to new pharmaceuticals or dietary interventions for the treatment of immobility osteopenia. PMID- 12744262 TI - A general framework for neurobiological modeling: an application to the vestibular system. AB - The otolith organs in the vestibular system are excellent detectors of linear accelerations. However, any measurement of linear acceleration is ambiguous between a tilt in a gravitational field and an inertial acceleration. Angelaki et al. have put forward a general hypothesis about how inertial accelerations can be computed based on vestibular signals (J. Neurosci. 19 (1999) 316). We have constructed a realistic, detailed model of the relevant systems to test this hypothesis. The model produces useful predictions about what kinds of neurons should be found in the vestibular nucleus if such a computation is actually performed in the vestibular system. The model is constructed using general principles of neurobiological simulation (J. Neurophys. 84 (2000) 2113). PMID- 12744263 TI - Biomedical applications of simulated environments. AB - Environmental physiology assumes great significance in our national context in view of the diverse climatic conditions prevailing in different regions. Troops have to operate in diverse environmental conditions guarding the frontiers. Hence, the research in this area has been focused on the usage of field studies in the natural environments or simulated environments in the laboratory. Besides, the application of the simulation chambers in the research on the physiological effects of diverse environments, these studies may have applications in the control and management of certain clinical disorders. Some simulation chambers and specialised set-ups have been designed and developed at the Defence Institute of Physiology and Allied Sciences to carry out simulation studies. This paper describes these developments and the potentials of these biomedical applications of simulated environments. PMID- 12744264 TI - So much to do, so little time: physician capacity constraints, 1997-2001. AB - Signs of tightened physician capacity--or physicians' ability to provide services relative to demand--appeared between 1997 and 2001, according to a study by the Center for Studying Health System Change (HSC). Patients waited longer for appointments, and more physicians reported having inadequate time with patients. Despite signs of tightened physician capacity, the supply of physicians grew modestly, the proportion of physicians working with nurse practitioners and other caregivers increased and doctors spent more time in direct patient care. This seeming contradiction emerged as the retreat from tightly managed care gave patients freedom to seek more care without substantial out-of-pocket cost increases. Current physician capacity constraints may ease if higher out-of pocket costs prompt patients to seek less care. PMID- 12744265 TI - Decision making in the NICU--strategies, statistics, and "satisficing". AB - As science progresses, new issues in bioethics grab the headlines, but in neonatal medicine communication and decision making for sick and premature infants is as important today as it was thirty years ago. Decisions have to be made and made well, despite suboptimal time, data, or knowledge. To this end, the authors propose good communication and a cooperative model as key to interpreting the best interest standard in a way that respects parental rights and responsibility and allows best interest to be decided from an amalgam of medical facts and human values. PMID- 12744267 TI - To show our humanness--relational and communicative competence in pediatric palliative care. AB - The death of a child is perhaps the worst tragedy a family ever has to endure. The communication that occurs among children, parents, and healthcare professionals at the end of a child's life must be grounded in caring, and compassionate relationships. These relationships require particular skills, knowledge and attitudes that are not fully addressed in many approaches to communication training currently available to practitioners. This essay proposes elements of a pedagogy for relational and communicative competence in pediatric palliative care that is rooted in ethical and ethnographic principles. PMID- 12744266 TI - Facing decisions about life and death--communication with parents. AB - How can healthcare professionals, most of whom have never personally been confronted with such tragic situations, best help parents and their children make decisions about life and death that will minimize the suffering of children and their parents? Research studies and the professional experience of those attuned to these problems can be instructive. This paper reviews and gives advice about the communication between healthcare professionals and parents and children facing life and death. PMID- 12744268 TI - Children's participation in healthcare in the UK--gesture, rhetoric, or real involvement? AB - In the United Kingdom (England, Wales, Scotland, and Northern Ireland) children and their best interests are protected through a range of best practice initiatives, and legislation and guidance at country, national, European, and global levels. Some of the recent commitment by the government may be the result of enlightened thinking, but some of it has resulted from the aftermath of at least two major healthcare incidents. This article reviews the UK's recent national and international efforts to protect its thirteen million children and ensure that their voices are heard. PMID- 12744269 TI - Child assent and parental permission for clinical research--some considerations. AB - The success of our future efforts to understand and improve the ethics of pediatric informed consent may depend, in large measure, on our willingness and ability to conceive of child assent and parental permission as joint, mutually affective processes. Given current trends, our empirical efforts may need to unfold at the interface of assent and parental permission, rather than exclusively or even primarily in one domain or the other. This shift will permit researchers to identify those areas in which the two mechanisms function in concert--in the best interests of patients, parents, and clinicians--and those in which they do not. Targeting these problematic areas for intervention and improvement may result in a more effective consent process for clinical research involving minors. PMID- 12744270 TI - Confidentiality for mental health concerns in adolescent primary care. AB - Guidelines from several national professional groups and a patchwork of state laws support the option to provide confidential mental healthcare for adolescents as a way to reduce barriers to treatment. These guidelines do not, however, help doctors decide when and to what extent confidentiality might be appropriate. We propose a set of practical considerations that clinicians can use to develop and justify confidentiality and family involvement in individual cases. Use of this framework may increase clinician comfort in discussing confidentiality and mental health topics with adolescents, and thus reduce barriers to the management of mental health problems in adolescent primary care. PMID- 12744271 TI - Ethical concerns associated with childhood depression. AB - Children always merit special ethical concern in mental health research and treatment. But children with depression are a particularly vulnerable population because of developmental considerations and the severity of the illness. This article reviews ethical concerns regarding assessment of depression, clinical care, and research with children. PMID- 12744272 TI - Case study: James vs. the Board of Education. PMID- 12744274 TI - Educational programs for deaf students. Schools and programs in the U.S. PMID- 12744275 TI - Educational programs for deaf students. Schools and programs in Canada. PMID- 12744276 TI - Educational programs for deaf students. Postsecondary programs. PMID- 12744273 TI - Elements of a neurobiological theory of the hippocampus: the role of activity dependent synaptic plasticity in memory. AB - The hypothesis that synaptic plasticity is a critical component of the neural mechanisms underlying learning and memory is now widely accepted. In this article, we begin by outlining four criteria for evaluating the 'synaptic plasticity and memory (SPM)' hypothesis. We then attempt to lay the foundations for a specific neurobiological theory of hippocampal (HPC) function in which activity-dependent synaptic plasticity, such as long-term potentiation (LTP), plays a key part in the forms of memory mediated by this brain structure. HPC memory can, like other forms of memory, be divided into four processes: encoding, storage, consolidation and retrieval. We argue that synaptic plasticity is critical for the encoding and intermediate storage of memory traces that are automatically recorded in the hippocampus. These traces decay, but are sometimes retained by a process of cellular consolidation. However, we also argue that HPC synaptic plasticity is not involved in memory retrieval, and is unlikely to be involved in systems-level consolidation that depends on HPC-neocortical interactions, although neocortical synaptic plasticity does play a part. The information that has emerged from the worldwide focus on the mechanisms of induction and expression of plasticity at individual synapses has been very valuable in functional studies. Progress towards a comprehensive understanding of memory processing will also depend on the analysis of these synaptic changes within the context of a wider range of systems-level and cellular mechanisms of neuronal transmission and plasticity. PMID- 12744277 TI - University and college programs for personnel in deafness. PMID- 12744278 TI - Programs for deaf-blind children and adults. PMID- 12744279 TI - Programs and services for the deaf. Supportive and rehabilitative programs. PMID- 12744280 TI - Research on deafness. Doctoral dissertations. PMID- 12744281 TI - Avoidance of hidden allergens in processed foods: a challenge for food chemists and manufacturers. PMID- 12744282 TI - Studies on dimerisation of tocopherols under the influence of methyl linoleate peroxides. AB - The stability of plant oils is related to the level of polyunsaturated fatty acids and the presence of native antioxidants--especially tocopherols. During storage, lipids or the fat products undergo oxidation and tocopherol dimers and trimers are formed. These compounds possess reducing and antioxidant properties and participate in oxidation clearly inhibiting this process. In the present study, the correlation between levels of peroxides formed during autoxidation of methyl linoleate and simultaneous decomposition of tocopherols was examined. The peroxide value was investigated. Quantities of decomposed tocopherols and formation of their dimers were determined by high-performance liquid chromatography (HPLC). Mass spectrum analysis confirmed thatthe analysed compounds were dimes. Dimerisation of gamma-T begins at the smaller quantity of the methyl linoleate peroxides than dimerisation of delta-T. At the beginning of methyl linoleate autoxidation dimerisation of gamma-T in relation to its loss was smaller. The quantity of gamma-T dimers with ether bonds in total dimers pointed to faster binding of phenoxy radicals than transformation into the phenyl ones. delta-T dimers with phenyl bonds constitute about 65% of the total. The quantity of peroxides in methyl linoleate, necessary for quantitative and qualitative changes of homologous tocopherols, decreased from delta-T to alpha-T. PMID- 12744283 TI - Antioxidant properties of commercial alcoholic and nonalcoholic beverages. AB - Recent interest in food phenolics has increased greatly, because of their antioxidant and free radical scavenging abilities. Popular beverages in the world include tea, coffee, cocoa, beer, wine and fruit/vegetable juices. All of these beverages contain phenolic compounds. In present study total polyphenol content and in vitro antioxidant properties were investigated in 16 red wines, 5 white wines, 5 lager beers, 3 dark beers, 17 fruit juices and 5 vegetable juices. High polyphenol content was measured in red wines (1720 +/- 546 mg x L(-1)) and in some fruit juices such as elderberry and prunes (5,680 and 1,807 mg x L(-1), respectively). The concentration of polyphenols was between 159 and 5,680 mg x L( 1) in fruit juices and between 255 and 696 mg x L(-1) in vegetable ones, while low level of phenolics was observed in dark and lager beers and white wines (473, 376 and 392 mg x L(-1), respectively). All samples exhibited significant antioxidant properties such as hydrogen-donating ability, reducing power, chelating ability and total antioxidant status (TAS) value. These antioxidant properties strongly correlated with the total polyphenol content of the beverages. PMID- 12744284 TI - In vitro study on digestion of peptides in Emmental cheese: analytical evaluation and influence on angiotensin I converting enzyme inhibitory peptides. AB - A simple in vitro protocol simulating gastrointestinal digestion of proteins and peptides to investigate the effect of digestive enzymes on the biological activity of peptides present in dairy products was developed. This protocol consisted in a 30 min incubation with pepsin followed by a 4 h incubation with trypsin or pancreatin. It was applied to an Emmental cheese water-soluble extract (WSE) and to a casein solution (as a control). Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) allowed to monitor the digestion of proteins. Reversed-phase high-performance liquid chromatography (RP-HPLC) allowed to monitor the conversion of proteins and peptides into peptides and amino acids: it is proposed to use the mean retention time corresponding to the overall retention time distribution of molecules to assess the effect of digestive enzymes. The biological activity focused in this study was the angiotensin I converting enzyme (ACE) inhibitory activity. Digestion of Emmental WSE induced an increase of the ACE inhibition as compared to undigested WSE while a 10 kDa ultrafiltered WSE lost a part of its ACE inhibitory activity after digestion process. These results strongly suggest that digestive enzymes diminished the ACE inhibition by the peptides present in Emmental cheese WSE, while the digestion of peptides of high molecular weight would generate new ACE inhibitory peptides. PMID- 12744285 TI - Nutritional value of broad bean seeds. Part 1: Starch and fibre. AB - This paper characterizes starch and dietary fibre in the seeds of fine-size and large-size seed varieties of broad bean and in edible varieties of pea for comparison. The experimental material included seeds at full physiological maturity. The fine-size seed varieties of broad bean, Gobik and Goral, contained slightly less total starch (62.32% and 62.19% of dry matter) than the large-size seed varieties, Windsor Bialy and Bartom (65.32% and 65.26% of dry matter). It was shown that fine-size seed varieties of broad bean are a good source of resistant starch, which is comparable to large-size seed varieties. As far as the content of total starch is concerned, the share of this form of starch in broad bean seeds amounted to 1/3. Digestible starch in broad beans was in the range of 39-42% of dry matter, pea contained approximately one half less starch of this type. Moreover, it was found that the relation of rapidly digestible starch to slowly digestible starch amounted to 1:1 irrespective of broad bean variety. Dietary fibre in broad beans was in the range of 20.36%-26.79% of dry matter while half of it was found to be concentrated in the seed coat. The soluble fraction of the total content of fibre amounted to 11.81% in the Bartom variety and up to 15.89% in the Gobik variety. PMID- 12744286 TI - Nutritional value of broad bean seeds. Part 2: Selected biologically active components. AB - Selected biologically active non-nutrient components (BANS) were determined in broad bean seeds of fine-size and large-size seed varieties and in pea seeds for comparison. Among the analysed biologically active substances the broad bean seeds of fine-size and large-size seed varieties were differentiated mainly by phenolic compounds (including flavanols and proantocyanidines) which appear in twice as large quantities in large-size seed varieties. It was shown that in comparison to pea, broad bean seeds are characterised by a higher content of phenolic compounds, phytates as well as a higher activity of inhibitors of trypsin and amylases. Moreover, it was found that phenolic compounds accumulate mainly in the dark-colored seed coats of large-size broad bean and this fact is related to higher activity inhibiting-amylases of methanol extracts from this fraction of seeds. PMID- 12744287 TI - Studies on organoleptic properties of food products from fresh egg and egg powder through principal component analysis. AB - The population which is below the poverty line is devoid of nutritious diet. Egg and milk are categorized as complete foods. The defensive organizations are situated in such remote places where fresh food material is not available. Keeping in view these problems, the study of organoleptic variables, viz., color, appearance, aroma, texture and taste in the food products of cake, omelet doughnut, coconut macaroon and mayonnaise from fresh egg and egg powder, was conducted. Principal component analysis was carried out. Organoleptic properties of doughnut prepared from egg powder were superior compared to fresh egg which had better sensory traits for coconut macaroon. The sensory traits like taste, texture and aroma were the most influential traits studied to pronouncing as a panel decision. It is proposed that fresh egg and egg powder should be preferred in the process of preparation of coconut macaroon and doughnut, respectively. PMID- 12744288 TI - Use of starch/xanthan gum combinations as thickeners of cocoa syrups. AB - The influence of starch-xanthan thickening systems on the sensory, textural and rheological properties of cocoa syrups was studied. The results of whole and partial sensory analyses of syrups obtained under laboratory conditions were presented and compared to those of commercially available syrups. In the evaluation of texture such parameters as hardness, adhesiveness, cohesiveness, gumminess and stringiness were considered. The results of rheological measurements were presented in the form of flow curves as well as viscosity/time and viscosity/ temperature relationships. Flow curves were fitted to the models of Ostwald-de Waele and Casson, which allowed characterising the syrups under study to non-Newtonian, pseudoplastic, and thixotropic fluids. It was concluded that the quantity of thickeners used influenced not only theological, but also textural and sensory properties of the syrups. The decrease of pseudoplasticity and hardness with an increase of xanthan addition was a result of high sugars level in the syrups. This phenomenon was a result of competition between sugars (mono-, di-, and oligosaccharides) and hydrocolloids for water molecules in the systems under study. PMID- 12744289 TI - Studies on the amino acid and mineral content of buckwheat protein fractions. AB - Protein fractions were extracted by successive extraction and analysis method in four buckwheat varieties including Japanese spring buckwheat, Japanese summer buckwheat, Yuqiao No. 1 and Yuqiao 6-21. The amino acid and the mineral content of each protein fraction were also analyzed in this paper. The basis for the data on the experiment was the relationship between protein fractions and amino acids, and the mineral contents of protein fractions. The distribution and proportion of amino acids and the minerals in each protein fraction in the buckwheat kernel as well as the nutritional value of buckwheat kernel were discussed. The results showed that there is a high amount of soluble protein and very low amount of prolamin in the buckwheat kernel. Leucine is the first limited amino acid in buckwheat, and buckwheat protein is only a seminutritional protein. The buckwheat kernel is rich in K, Zn in the albumin, Ca, Mg and Mn in the globulin, Na in the prolamin and glutelin. PMID- 12744290 TI - Antifungal activity of peppermint and sweet basil essential oils and their major aroma constituents on some plant pathogenic fungi from the vapor phase. AB - The vapors of peppermint oil and two of its major constituents (menthol and menthone), and sweet basil oil and two of its major constituents (linalool and eugenol), were tested against Sclerotinia sclerotiorum (Lib.), Rhizopus stolonifer (Ehrenb. exFr.) Vuill and Mucor sp. (Fisher) in a closed system. These fungi cause deterioration and heavy decay of peach fruit during marketing, shipping and storage. The essential oils, their major individual aroma constituents and blends of the major individual constituents at different ratios inhibited the growth of the fungi in a dose-dependent manner. Menthol was found to be the individual aroma constituent responsible for the antifungal properties of peppermint essential oil, while menthone alone did not show any effect at all doses. In the case of basil oil, linalool alone showed a moderate antifungal activity while eugenol showed no activity at all. Mixing the two components in a ratio similar to their concentrations in the original oil was found to enhance the antifungal properties of basil oil indicating a synergistic effect. PMID- 12744291 TI - Influence of modified atmosphere packaging on the shelf life of prebaked pizza dough with and without preservative added. AB - The possible effect of different modified atmospheres on the shelf life of prebaked pizza dough, with and without added calcium propionate, was investigated. Three packaging atmospheres were tested: 20% CO2: 80% N2, 50% CO2: 50% N2, 100% CO2, and air (control). Samples were examined daily for visible mold growth and analysed after 2, 8, 17 and 31 days throughout storage (15-20 degrees C and 54-65% relative humidity, RH) for changes in gaseous composition, pH and microbial populations (mesophilic aerobic and anaerobic bacteria, lactic acid bacteria (LAB), and yeasts and molds). Microbiological results showed that molds had a greater sensitivity to CO2 than bacteria and yeasts. Products containing calcium propionate did not show mold growth throughout storage (31 days) when packaged in air or in CO2-enriched atmospheres (20, 50 and 100%). However, in pizza dough without preservative (calcium propionate), mold growth was evident after 7 days, except under 100% CO2 atmosphere (13 days) regardless of the packaging atmosphere. From these results we conclude that the addition of calcium propionate had more and decisive influence on the shelf life extension of prebaked pizza dough. PMID- 12744293 TI - Thermal kinetics of color degradation of mulberry fruit extract. AB - The effects of temperature and pH on color degradation kinetics of the mulberry fruit extract were investigated. The absorbance at 510 nm was decreased with increase of heating time, but that at 420 nm was increased with the increase of heating time at 100 degrees C. The change of the browning index (A510/A420) was increased with increase of pH and was lower at pH 2.0 than that at pH 5.0. The browning index variation was adequately described by both the first-order and the zero-order kinetic. However, the zero-order kinetic model was proposed because of the better fit. According to the Arrhenius model, the activation energies for the browning index in the range of 80-100 degrees C for the four different pH values were 30.68 kJ/mol for pH 2.0, 35.87 kJ/mol for pH 3.0, 42.67 kJ/ mol for pH 4.0, and 43.49 kJ/mol for pH 5.0. PMID- 12744292 TI - In vitro evaluation of antioxidant properties of Cocos nucifera Linn. water. AB - Tender Cocos nucifera L. (Palmacea) water (CW), variety Chandrasankara, was tested for its ability to scavenge free radicals, inhibit lipid peroxidation and protect hemoglobin from nitrite-induced oxidation. Fresh sample of CW scavenged 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenz-thiazoline-6 sulfonic acid) (ABTS) and superoxide radicals but promoted the production of hydroxyl radicals and increased lipid peroxidation. The activity was most significant for fresh samples of CW and diminished significantly upon heat, acid or alkali treatment or dialysis. Maturity of coconut drastically decreased the scavenging ability of CW against DPPH, ABTS and superoxide radicals. CW protected hemoglobin from nitrite-induced oxidation to methemoglobin when added before the autocatalytic stage of the oxidation. Acid, alkali or heat treated or dialyzed CW showed a decreased ability in protecting hemoglobin from oxidation. The scavenging ability and protection of hemoglobin from oxidation may be partly attributed to the ascorbic acid, which is an important constituent of CW. As CW is a rich source of vitamins, amino acids and enzymes, etc., more than one active principle maybe involved. PMID- 12744294 TI - Rapid differentiation of new apple cultivars by headspace solid-phase microextraction in combination with chemometrical data processing. AB - The aim of this study was to test a combination of automated headspace solid phase-microextraction gas chromatography (GC) with chemometrical data treatment for the rapid differentiation of enzyme-inactivated homogenates of new apple cultivars. The four cultivars Pinova, Piflora, Renora and Florina are characterized by different volatile patterns. Differences in the contents of volatiles were especially found for butyl acetate, ethyl butanoate, 2-methyl butanol, ethyl acetate and 6-methyl-5-hepten-2-ol. The used sample preparation method for GC coupled with pattern recognition of chromatograms is a useful tool for rapid and reliable determination of large numbers of samples. PMID- 12744295 TI - Comparison of the quality of some Croatian and German wheat varieties according to the German standard protocol. AB - Ten Croatian and five German wheat varieties were tested. They were carried out using the German standard testing protocol. The results demonstrated that the quality of the Croatian wheat varieties could be ranged in A-C quality groups according to the German testing protocol. Croatian wheat varieties were comparable in many parameters (protein content, hardness, water absorption, dough handling properties and volume yield) with chosen German wheat varieties. Some differences were found in falling number, sedimentation value, flour yield, and ash value number. PMID- 12744296 TI - Protein composition of some Croatian and German wheat varieties and their influence on the loaf volume. AB - Ten Croatian and five German wheat varieties were tested. Wheat protein composition was analysed according to a modified Osborne fractionation. They were extracted in four fractions: NaCl fraction, ethanol fraction, acetic acid fraction and insoluble fraction. The influence of protein and protein composition on the loaf volume was investigated. In protein composition a difference between the Croatian and German wheat varieties were found. Protein content, glutenin content and insoluble glutenin content showed influences on the loaf volume. PMID- 12744297 TI - Lansoprazole overutilization: methods for step-down therapy. AB - OBJECTIVE: To identify the documented indications for long-term therapy with lansoprazole 30 mg twice daily at the Veterans Affairs Pittsburgh Healthcare System, assess compliance with appropriate use criteria, evaluate patients eligible for step-down therapy, and recommend appropriate step-down therapy in order to improve patient care, decrease overprescribing, and reduce medication costs. STUDY DESIGN: Prospective intervention. METHODS: The records of all patients with prescriptions for lansoprazole 30 mg twice daily as of June 2000 were reviewed. Patients were interviewed to assess medication compliance and symptom control and to provide education on lifestyle modifications. Interventions with the providers were completed to encourage step-down therapy in appropriate patients. RESULTS: Two hundred forty-eight patients with active prescriptions for twice-daily lansoprazole were reviewed. Of these patients, 66% (n = 163) did not have an indication compliant with the medical center's guidelines for use of lansoprazole 30 mg twice daily. Of these, 88% (n = 143) had no documented attempt at step-down therapy and 49% (n = 80) had no documented gastrointestinal workup. Interventions for step-down therapy were recommended for 48% (n = 120) of the 248 patients. Forty-six percent (n = 60) of recommendations were accepted, resulting in a cost savings of dollars 85000 per year. CONCLUSIONS: A high rate of clinician noncompliance with the guidelines for appropriate use of lansoprazole 30 mg twice daily was found. These prescribing patterns resulted in significant cost concerns. Our review and interventions led to step-down therapy for almost half of the patients receiving twice-daily lansoprazole. This review of patient records and intervention with primary care providers resulted in cost reduction and offered an opportunity to educate patients on beneficial lifestyle modifications. PMID- 12744298 TI - Reducing costs and improving outcomes of percutaneous coronary interventions. AB - OBJECTIVE: To describe cost reduction and quality improvement efforts in our percutaneous coronary intervention (PCI) program and how risk adjustment was used to assess the effects of these changes. STUDY DESIGN: Single center registry analysis. PATIENTS AND METHODS: Data were collected on 2158 PCIs performed between July 1, 1994, and June 30, 1997. Of these, 1126 PCIs reflected care provided after implementation of competitive bidding for catheterization lab supplies, and efforts to reduce the use of postprocedure heparin and to implement early arterial sheaths removal (postbidding period). Hospital costs were estimated using a microcost accounting method. In-hospital mortality rates during the 2 time periods were compared using standardized mortality ratio estimated with a previously validated risk adjustment model for in-hospital mortality. RESULTS: Compared with the prebidding period, the postbidding period was characterized by a significantly higher utilization of new technology (coronary stents and atherectomy devices 46% vs 25%; abciximab 19.1% vs 3.7, P<.01), and an overall increase in case complexity. Despite these changes, the average and median postbidding cost per case was dollars 1223 and dollars 1444 lower, respectively, than in the prebidding period. After adjustment for comorbidities, procedure variables, complications, and length of hospital stay, multivariate regression modeling identified the postbidding period as an independent predictor of lower hospital costs (P<.001) with an estimated adjusted cost savings of dollars 460. These cost savings were associated with trends toward a lower observed mortality rate, a higher predicted mortality rate, and a significantly lower standardized mortality ratio (SMR .71; 95% CI 0.48-0.9; P<.05). CONCLUSION: Despite an increase in case complexity and utilization of new technology, cost reductions can be achieved through competitive bidding for supplies and modifications of periprocedure care. Risk adjustment appears to be a valid tool for assessing the effectiveness of these efforts independently from changes in case mix. PMID- 12744299 TI - Evaluation of a depression health management program to improve outcomes in first or recurrent episode depression. AB - OBJECTIVES: To evaluate the impact of telephone counseling and educational materials on medication adherence and persistency among members with newly diagnosed depression enrolled in a pharmacy benefit management-sponsored disease management program. STUDY DESIGN: Longitudinal cohort observation. METHODS: The study population comprised 505 members with a new or recurrent episode of depression who consented and enrolled in a depression disease management program. After written consent was obtained, program participants received up to 4 telephone-counseling calls and 5 educational mailings focused on the importance of medication compliance, barriers to medication compliance, quality of life, symptoms, and satisfaction with the program. A control group of 3744 members was selected from client companies that opted not to offer the depression program. Measures of medication adherence, persistency with prescription drug therapy, and patient refill timeliness were computed for both groups and compared. RESULTS: Patients enrolled in the depression disease management program were significantly more likely to adhere to their medication regimen during acute (89.0% vs 67.7%, P < .001) and continuation treatment phases (81.1% vs 57.6%, P < .001). In addition, members enrolled in the program were significantly more likely to continue their therapy after 7 months (77.8% vs 49.5%, P < .001) and refilled their prescriptions on a more timely basis (0 vs 18 days, P < .001). CONCLUSIONS: A pharmacy benefit management-sponsored health management depression program succeeded in encouraging patients with new or recurrent depression to stay on antidepressant medication and to reach treatment goals outlined by best practice guidelines. PMID- 12744300 TI - A prediction model for targeting low-cost, high-risk members of managed care organizations. AB - OBJECTIVE: To describe the development and validation of a predictive model designed to identify and target HMO members who are likely to incur high costs. STUDY DESIGN: Split-sample multivariate regression analysis. PATIENTS AND METHODS: We studied enrollees in a 350000-member HMO with > or = 1 claim in 1998 and 1999. The prediction model uses a combination of clinical and behavioral vaiables and 1998 and 1999 claims data. The prediction model was applied and used to rank low-cost patients (1998 cost < dollars 2000) according to their estimated probability of incurring costs > or = dollars 2000 in 1999. For prospective testing, we applied our models to data that are not available in advance. The same prediction model was applied to rank a different set of low-cost patients (1999 cost < dollars 2000) according to estimated probability of incurring costs > or = dollars 2000 in 2000. Because the predictions were used for disease management purposes, the outcomes of a randomly selected control group not intervened on for the disease management program was analyzed. The predictive accuracy of the model was tested by comparing the percentages of "targeted" vs all low-cost patients who incurred high costs in the subsequent year. RESULTS: Of the low-cost, top-ranked 1998 patients, 47.8% incurred high (> or = dollars 2000) medical expenses in 1999 vs 14.2% of randomly selected patients who were low cost in 1998. Of the top-ranked 1999 patients, 39.7% incurred high costs in 2000 vs 12.2% of the randomly selected low-ranked patients. CONCLUSIONS: The prediction model successfully identifies low-cost, high-risk patients who are likely to incur high costs in the next 12 months. PMID- 12744301 TI - Effect of off-label use of oncology drugs on pharmaceutical costs: the rituximab experience. AB - BACKGROUND: While the off-label use of oncology interventions is widespread, the factors influencing off-label use and the resultant influence on oncology drug expenditures are not well understood. STUDY DESIGN: To assess the indications for rituximab use, a retrospective review was undertaken at a single academic center between September 1998 and June 2001. METHODS: Patient diagnoses were linked to pharmacy records, and each administration of rituximab was classified as either on-label or off-label as defined by FDA-approved indications. The resultant utilization patterns were the foundation for a conceptual model designed to identify factors that influence off-label use of oncology-related therapeutics and forecast the effect of off-label use on aggregate oncology drug expenditures. RESULTS: One hundred one patients received a total of 428 rituximab administrations during the study period. Most (320, 75%) of the administrations were for off-label indications. Although the extent of off-label and on-label use grew at a similar rate initially, off-label utilization increased nearly exponentially over time as on-label uses lessened. A conceptual model that describes factors that promote, inhibit, or have a mixed influence on off-label use may help predict future patterns of off-label utilization and allow better forecasting of oncology drug expenditures. CONCLUSIONS: The off-label use of rituximab is substantial. Projections of oncology-related patterns of care and drug expenditures must account for the potential for off-label use. PMID- 12744302 TI - Different Kv2.1/Kv9.3 heteromer expression during brain and lung post-natal development in the rat. AB - The Kv2.1/Kv9.3 heteromer generates an O2 sensitive potassium channel and induces a slow deactivation that has important consequences for brain and lung physiology. We examined the developmental regulation of Kv2.1 and Kv9.3 mRNAs in brain and lung. Both genes followed parallel expression patterns in brain, increasing progressively through post-natal life. In lung, however, the expression of the two genes followed opposite trends: Kv2.1 transcripts decreased, while Kv9.3 mRNA increased. The Kv9.3/Kv2.1 ratio shows that while in brain the expression of both genes followed a similar pattern, the relative abundance of Kv9.3 increased steadily through post-natal life in lung. Furthermore, there is selective regulation of gene expression during the suckling weaning transition. Our results suggest that different Kv9.3/Kv2.1 ratios could have physiological implications in both organs during post-natal development, and that diet composition and selective tissue-specific insulin regulation modulate the expression of Kv2.1 and Kv9.3. PMID- 12744304 TI - CHO intake alters obesity risk associated with Pro12Ala polymorphism of PPARgamma gene. PMID- 12744305 TI - Chemistry of materials under extreme high pressure-high-temperature conditions. AB - Most of our knowledge of chemistry is derived from experiments carried at the Earth's surface, at pressures near one atmosphere. However, most elements and compounds in the universe exist under conditions of extremely high pressures, often combined with high temperatures, deep within the planets and stars. Under these conditions, new high-density crystal forms occur, species usually known only as molecules become dense covalent or ionic solids, and insulators and semiconductors become metals and even superconductors. Valency states and coordination numbers are changed, and it is expected that chemical bonding and reactivity is modified. Paul McMillan describes how the field of condensed matter chemistry under extreme high pressure conditions now represents a vast new area to be explored. PMID- 12744306 TI - Chemical synthesis of magnetic nanoparticles. AB - Recent advances in the synthesis of various magnetic nanoparticles using colloidal chemical approaches are reviewed. Typically, these approaches involve either rapid injection of reagents into hot surfactant solution followed by aging at high temperature, or the mixing of reagents at a low temperature and slow heating under controlled conditions. Spherical cobalt nanoparticles with various crystal structures have been synthesized by thermally decomposing dicobalt octacarbonyl or by reducing cobalt salts. Nanoparticles of Fe-Pt and other related iron or cobalt containing alloys have been made by simultaneously reacting their constituent precursors. Many different ferrite nanoparticles have been synthesized by the thermal decomposition of organometallic precursors followed by oxidation or by low-temperature reactions inside reverse micelles. Rod-shaped iron nanoparticles have been synthesized from the oriented growth of spherical nanoparticles, and cobalt nanodisks were synthesized from the thermal decomposition of dicobalt octacarbonyl in the presence of a mixture of two surfactants. PMID- 12744303 TI - Nucleoside transporters in absorptive epithelia. AB - There are two families of nucleoside transporters, concentrative (termed CNTs) and equilibrative (called ENTs). The members of both families mediate the transmembrane transport of natural nucleosides and some drugs whose structure is based on nucleosides. CNT transporters show a high affinity for their natural substrates (with Km values in the low micromolar range) and are substrate selective. In contrast, ENT transporters show lower affinity and are more permissive regarding the substrates they accept. Both types of transporters are tightly regulated in all cell types studied so far, both by endocrine and growth factors and by substrate availability. The degree of cell differentiation and the proliferation status of a cell also affect the pattern of expressed transporters. Although the presence of both types of transporters in the cells of absortive epithelia suggested the possibility of a transepithelial flux of nucleosides, their exact localization in the different plasma membrane domains of epithelial cells had not been demonstrated until recently. Concentrative transporters are found in the apical membrane while equlibrative transporters are located in the basolateral membrane, thus strengthening the hypothesis of a transepithelial flux of nucleosides. PMID- 12744307 TI - Phase-transfer alkylation reactions using microreactors. AB - Phase-transfer alkylation in a microreactor proceeds smoothly, and the reaction has been found to be more efficient than that in a round-bottomed flask with vigorous stirring; we have observed by an optical microscope study that an interfacial area provided by organic and aqueous phases is more extended in a microreactor. PMID- 12744308 TI - Bronsted acid-base ionic liquids and their use as new materials for anhydrous proton conductors. AB - Novel Bronsted acid-base ionic liquids, derived from a simple combination of a wide variety of organic amines with bis(trifluoromethanesulfonyl) amide are electroactive for H2 oxidation and O2 reduction at a Pt electrode under non humidifying conditions, which shows the prospect of the use of protic ionic liquids as new materials for anhydrous proton conductors at elevated temperatures. PMID- 12744309 TI - Anion-sealed single-molecule capsules. AB - A new approach to anion recognition utilizing electrostatic and hydrogen bonding interactions has been demonstrated by placement of the whole ion-pair in a molecular capsule. PMID- 12744310 TI - Optimizing the reversibility of hydrazone formation for dynamic combinatorial chemistry. AB - Hydrazones from hydrazines bearing electron withdrawing groups, and aromatic or aliphatic aldehydes form and hydrolyse rapidly in water at neutral pH. PMID- 12744311 TI - A xylobiose-derived isofagomine lactam glycosidase inhibitor binds as its amide tautomer. AB - The atomic-resolution structure of a xylobiose-derived isofagomine lactam in complex with the xylanase Xyn10A from Streptomyces lividans reveals that the lactam is bound to the enzyme as the amide tautomer, with "reversed" protonation states for nucleophile and acid-base. PMID- 12744313 TI - C-c bond formation through oxidatively induced elimination of platinum complexes- a novel approach towards conjugated macrocycles. AB - A novel and effective method for the synthesis of conjugated macrocycles is described. By the self-assembly of conveniently accessible building blocks to a metalla-macrocycle, and subsequent C-C bond formation through elimination of transition metal units, the strained cyclodimeric terthiophene-diyne 4 -as a precursor for cyclo[8]thiophene 5- was synthesized in a good overall yield. PMID- 12744312 TI - Distortion of a cellobio-derived isofagomine highlights the potential conformational itinerary of inverting beta-glucosidases. AB - A cellobio-derived isofagomine glycosidase inhibitor (Ki approximately 400 nM) displays an unusual distorted 2,5B (boat) conformation upon binding to cellobiohydrolase Cel6A from Humicola insolens, highlighting the different conformational itineraries used by various glycosidases, with consequences for the design of therapeutic agents. PMID- 12744314 TI - Oligonucleosides with a nucleobase-including backbone; synthesis and self association of novel dinucleotide analogues. AB - The synthesis and self-association of protected oxymethylene-bridged UA analogues are described. PMID- 12744316 TI - Electrochemically induced ring-closing of photochromic 1,2 dithienylcyclopentenes. AB - An unprecedented combination of photochromism and electrochromism is observed for two 1,2-bis(dithienyl)cyclopentene derivatives; the ring-opening reactions are photochemically driven while the ring-closing reactions can be triggered by electrochemical oxidation. PMID- 12744317 TI - One- and three-dimensional infinite arrays of Cu(I) ions exhibited by [Cu(NH3)2]Br and [Cu(NH3)Cl] in the solid state. AB - In the solid state, [Cu(NH3)Cl] forms a three-dimensional network with each Cu(I) ion being surrounded by three other Cu(I) centres in a trigonal-planar fashion [Cu...Cu = 2.979(1) A; cubic space group I2(1)3], whereas in [Cu(NH3)2]Br the cations establish infinite linear (Cu...Cu) chains spanning the crystal lattice [Cu...Cu = 2.931(1) A; monoclinic space group C2/c]. PMID- 12744315 TI - Cyclopentanes from N-amino-glyconolactams. A synthesis of mannostatin A. AB - Oxidation of a N-amino-ribonolactam with lead tetraacetate yields two cyclopentanes; the major one was transformed into the alpha-mannosidase inhibitor mannostatin A. PMID- 12744318 TI - TEM stereo-imaging of mesoporous zeolite single crystals. AB - Mesoporous zeolite single crystals with intracrystalline mesopores and metal oxide particles located in the zeolite mesopore are characterised by direct TEM stereo-imaging. PMID- 12744319 TI - Highly enantioselective hydrolysis of alicyclic meso-epoxides with a bacterial epoxide hydrolase from Sphingomonas sp. HXN-200: simple syntheses of alicyclic vicinal trans-diols. AB - Hydrolysis of N-benzyloxycarbonyl-3,4-epoxy-pyrrolidine and cyclohexene oxide with the epoxide hydrolase of Sphingomonas sp. HXN-200, respectively, gave the corresponding vicinal trans-diols in high ee and yield, representing the first example of enantioselective hydrolysis of a meso-epoxide with a bacterial epoxide hydrolase. PMID- 12744320 TI - In situ magnetic resonance imaging of electrically-induced water diffusion in a nafion ionic polymer film. AB - By deploying a functioning electrochemical cell inside a Magnetic Resonance Imaging (MRI) instrument, images of the electrically-induced diffusion of water through a Li+ ion-exchanged Nafion ionic polymer film in the form of two dimensional maps of proton density and transverse relaxation time, T2, were generated and changes in these images over time and with respect to changes made to the applied potential were followed. PMID- 12744321 TI - Modified micro-space using self-organized nanoparticles for reduction of methylene blue. AB - By using silica layers bound to the surface of the inner wall of the microchannel via a self-assembly technique which relies on capillary forces to organize the colloids, TiO2 particles were attached to the inner surface of a microcapillary. The reduction rate of methylene blue increased by more than 150 times in the SiO2/TiO2 modified micro-space compared to that in a batch system. PMID- 12744322 TI - Fabrication of a stable inorganic-organic hybrid multilayer film with uniform and dense inorganic nanoparticle deposition. AB - A stable inorganic-organic hybrid multilayer film with homogeneous and dense inorganic nanoparticle deposition was constructed by coating ZrO2 nanoparticles with poly(4-sodium styrenesulfonate) (PSS) and irradiating multilayer film assembled from PSS-coated ZrO2 nanoparticles and a diazo-resin (DR). PMID- 12744323 TI - De novo design of non-hydrogen-bonded helical pseudopeptides composed of oxanipecotic acid oligomers. AB - Ab initio calculation and circular dichroism experiments reveal that Oxa oligomers adopted pronounced non-hydrogen-bonded helical structures. PMID- 12744324 TI - A new dark quencher for use in genetic analysis. AB - A novel, long-wavelength, non-fluorescent quencher (LQ), based on 1,4 diaminoanthraquinone, has been incorporated at the 3' and 5'-termini of oligonucleotides. The quencher has been used in Molecular beacons, efficiently quenching the long wavelength fluorophore, Cy5. PMID- 12744325 TI - A novel peptide synthesis using fluorous chemistry. AB - Three new fluorous supports for peptide synthesis, i.e., the trialkoxybenzhydryl type (6), the Wang-type (7) and the tert-butyl-type support (8), were prepared. A bioactive peptide TRH was easily synthesized by an Fmoc strategy using the benzhydryl-type fluorous support with fluorous chemistry. PMID- 12744326 TI - A free radical initiated optically active vinyl polymer with memory of chirality after removal of the inducing stereogenic center. AB - Free radical polymerization of (+)-2,5-bis[4'-((S)-2 methylbutyloxy)phenyl]styrene yields a chiral polymer with memory of optical activity after the initial stereogenic center in the side group of the monomer is chemically removed. PMID- 12744327 TI - Ag8Cl2[Se2P(OEt)2]6: A rare example containing a combination of discrete clusters and chains. AB - The novel halide-centered Ag(I)8 cubic clusters containing diethyl diselenophosphato ligands are prepared and their solid state structures, a discrete unit or a one-dimensional chain, are dictated by the counter anions. PMID- 12744328 TI - Glyco-helix: parallel lactose-lactose interactions stabilize an alpha-helical structure of multi-glycosylated peptide. AB - Glyco-helix is designed as a novel model system to investigate cis carbohydrate carbohydrate interactions. Adhesive Lac-Lac interactions stabilize alpha-helix of Lac-peptide in the presence of fluorinated alcohols, but no such an interaction was observed in GlcNAc-peptide. PMID- 12744330 TI - Polymer nanosphere lithography: fabrication of an ordered trigonal polymeric nanostructure. AB - A combination of nanosphere lithography, sintering rheology and selective dissolution of self-assembled colloidal arrays was used to fabricate a novel trigonal polymer nanostructure whose trigonal structure became more evident after Ar+ ion etching. PMID- 12744329 TI - Low-temperature synthesis of titanium disulfide nanotubes. AB - A low-temperature gas reaction was used to successfully synthesize TiS2 nanotubes with an outer diameter of approximately 20 nm, an inner diameter of approximately 10 nm, an interlayer spacing of approximately 0.57 nm, and an average length of 2 5 microm. PMID- 12744331 TI - In situ FTIR studies on the electrochemical reduction of halogenated phenols. AB - Electrochemical reduction of a variety of mono- and di-chloro- and bromo- phenols at a palladised titanium electrode afforded phenolate in all cases according to in situ FTIR studies, with the same intermediate species being observed in some cases. PMID- 12744332 TI - Water solubilization, determination of the number of different types of single wall carbon nanotubes and their partial separation with respect to diameters by complexation with eta-cyclodextrin. AB - Complexation of single-wall carbon nanotubes with 12-membered cyclodextrins enables not only their solubilization in water but also their partial separation with respect to diameters and determination of the number of nanotube types on the basis of NMR spectra. PMID- 12744333 TI - Diastereoselective formation of a dipalladium(I) complex supported by a bridging tetradentate ligand, and oxidative addition of RS-H across a phosphine-bridged Pd(I)-Pd(I) bond. AB - The ligand CH2(P(o-C6H4NMe2)2)2 (dmapm) gives access to the first example of a dipalladium(I) complex supported by a tetradentate ligand, Pd2Cl2(mu-N,P,P,N dmapm), which, unlike the well-known Pd2X2(mu-dppm)2 complexes (X = halide, dppm = bis(diphenylphosphino)methane), reacts with thiols to give addition of RS-H across a phosphine-bridged Pd-Pd bond. PMID- 12744334 TI - A novel type of phosphide: synthesis and X-ray crystal structure analysis of (tBu3Si)3P4Li3. AB - The tetraphosphides (tBu3Si)3P4M3 (M = Li, Na) and (tBu2PhSi)3P4Na3 have been synthesized in high yield from the reaction of 3 equivalents of the silanides tBu3SiM (M = Li, Na) and tBu2PhSiNa with P4 in benzene. (tBu3Si)3P4M3 (M = Li, Na) are transformed into the unsaturated triphosphides (tBu3Si)2P3M (M = Li, Na) and tBu3SiPM2 in tetrahydrofuran at ambient temperature. PMID- 12744335 TI - Thermolysis of fluorinated cycloalkylidene fulgides yields a new class of photochromic compounds. AB - A series of fluorinated cycloalkylidene indolylfulgides has been designed, synthesized and characterized; most of the thermolysis products of these fulgides maintain photochromicity and display outstanding thermal and photochemical stability. PMID- 12744336 TI - Homochiral 3D open frameworks assembled from 1- and 2-D coordination polymers. AB - Homochiral 3D open frameworks have been assembled from 1- and 2-D coordination structures via hydrogen bonding and are stable towards the removal of included guest molecules. PMID- 12744337 TI - A highly sensitive and selective fluorescent molecular sensor for Pb(II) based on a calix[4]arene bearing four dansyl groups. AB - A new fluorescent molecular sensor based on a calix[4]arene bearing four dansyl groups exhibits very efficient binding in acetonitrile-water for lead and the changes of emission properties allows a detection limit of 4 microg L(-1). PMID- 12744339 TI - The role of isomorphism in synthetic analysis. Pruning the search tree by finding disjoint isomorphic substructures. AB - Isomorphism is an equivalence relation that is less stringent than identity (equality), and it is useful for synthetic analysis, since it allows one to find reflexive routes for targets even when they do not have an element of symmetry. PMID- 12744338 TI - Water-soluble supramolecular bowls formed by intra-clipping of resorcin[4]arene based ligands with Pd(II) ions. AB - Bowl-shaped superstructures have been constructed by intra-clipping of resorcin[4]arene derivatives with two equivalents of (en)Pd(NO3)2 in water. The binding phenomena of the aromatic carboxylates were proven to be both enthalpically and entropically favoured. PMID- 12744340 TI - Direct observation of oxygen depletion and product formation during photocatalysis at a TiO2 surface using scanning electrochemical microscopy. AB - Using scanning electrochemical microscopy (SECM) we have measured quantitatively the depletion of O2 during the photodegradation of 4-chlorophenol at supported TiO2 films for the first time and established the connection between Cl-formation and O2 depletion rates. PMID- 12744341 TI - Multivalent recognition of bis- and tris-Zn-porphyrins by N-methylimidazole functionalized gold nanoparticles. AB - N-Methylimidazole-functionalized gold nanoparticles behave as multivalent ligands for porphyrin arrays with an increase in binding strength of up to three order of magnitude with respect to a monovalent system. PMID- 12744342 TI - A substituted triaza crown ether as a binding site in DNA conjugates. AB - Synthesis of an asymmetrically substituted triaza crown ether, its incorporation into the 3'-end and 5'-end of ninemer oligonucleotides, and the influence of various alkanediamine ligands on duplex thermostabilities are reported. PMID- 12744343 TI - High tetraalkylaluminate fluxionality in half-sandwich complexes of the trivalent rare-earth metals. AB - Steric factors govern the formation of half-sandwich complexes (C5Me4R)Ln[N(SiHMe2)2]2 according to acid-base reactions utilising Ln[N(SiHMe2)2)3(thf)2 and substituted cyclopentadienes. Subsequent trimethylaluminium-promoted silylamide elimination produces the first half sandwich bis(tetramethylaluminate) complexes (C5Me4R)Ln(AlMe4)2. PMID- 12744344 TI - Versatile synthesis of nanometer sized hollow silica spherest. AB - Using the controlled precipitation of silicic acid on functionalized polystyrene latexes, nanometer sized silica-coated spheres could be prepared and subsequently modified to allow dispersion in non-aqueous solvents; removal of the interior polymer by calcination resulted in the formation of hollow silica spheres. PMID- 12744346 TI - Selective synthesis of conformationally restricted mono-cyclopentadienyl titanium(IV) complexes of p-tBu-calix[6]arene. AB - Reaction of p-'Bu-calix[6]arene with potassium metal in methanol followed by [TiCp2Cl2] affords novel mononuclear and binuclear monocyclopentadienyl titanium(IV) complexes, both having the same inverted double cone conformation with a Cp in one of the cavities, in both complexes. PMID- 12744345 TI - Self assembly, structure and properties of the decanuclear lanthanide ring complex, Dy10(OC2H4OCH3)30. AB - Self assembly produced high yields of the lanthanide ring complex Dy10(OC2H4OCH3)30, the largest lanthanide ring known, characterized by X-ray diffraction methods and by magnetic susceptibility as a function of temperature. PMID- 12744347 TI - First generation of pentazole (HN5, pentazolic acid), the final azole, and a zinc pentazolate salt in solution: A new N-dearylation of 1-(p-methoxyphenyl) pyrazoles, a 2-(p-methoxyphenyl) tetrazole and application of the methodology to 1-(p-methoxyphenyl) pentazole. AB - Ceric ammonium nitrate (CAN) in methanol-water gave a new N-dearylation of a series of substituted 1-(p-methoxyphenyl) pyrazoles and a 2-(p methoxyphenyl)tetrazole producing p-benzoquinone and the parent azole in a mole for mole ratio. Application of this reaction to 1-(p-methoxyphenyl) pentazole at 40 degrees C produced p-benzoquinone. 15N NMR spectra suggest that pentazole, HN5, was also produced and held in solution as N5- with Zn2+ ion. The 15N signal from N5- was -10.0 +/- 2.0 ppm in agreement with calculated values. PMID- 12744348 TI - A new type of three-dimensional framework constructed from dodecanuclear cadmium(II) macrocycles. AB - A new type of three-dimensional framework based on dodecanuclear Cd(II) macrocycles [[Cd2(dpa)(pya)]6(pya)6(dpe)3]n (1) (H2dpa = diphenic acid; Hpya = isonicotinic acid, dpe = 1,2-di(4-pyridyl)ethylene) was prepared by the hydrothermal reaction and in situ synthesis of pya from dpe precursor. PMID- 12744349 TI - Deep brain stimulation for obsessive-compulsive disorder. PMID- 12744350 TI - Hyperoxia: good or bad? PMID- 12744351 TI - Guglielmi detachable coil technology. PMID- 12744352 TI - Cushing disease. PMID- 12744354 TI - Guglielmi detachable coil embolization of cerebral aneurysms: 11 years' experience. AB - OBJECT: The authors report on their 11 years' experience with embolization of cerebral aneurysms using Guglielmi Detachable Coil (GDC) technology and on the attendant anatomical and clinical outcomes. METHODS: Since December 1990, 818 patients harboring 916 aneurysms were treated with GDC embolization at University of California at Los Angeles Medical Center. For comparative purposes, the patients were divided into two groups: Group A included their initial 5 years' experience with 230 patients harboring 251 aneurysms and Group B included the later 6 years' experience with 588 patients harboring 665 aneurysms. Angiographically demonstrated complete occlusion was achieved in 55% of aneurysms and a neck remnant was displayed in 35.4% of lesions. Incomplete embolization was performed in 3.5% of aneurysms, and in 5% occlusion was attempted unsuccessfully. A comparison between the two groups revealed a higher complete embolization rate in patients in Group B compared with that in Group A patients (56.8 and 50.2%, respectively). The overall morbidity/mortality rate was 9.4%. Angiographic follow ups were obtained in 53.4% of cases of aneurysms, and recanalization was exhibited in 26.1% of aneurysms in Group A and 17.2% of those in Group B. The overall recanalization rate was 20.9%. Note that recanalization was related to the size of the dome and neck of the aneurysm. Overall incidence of delayed aneurysm rupture was 1.6%, a rate that improved in the past 5 years to 0.5%. Ten of 12 delayed ruptures occurred in large or giant aneurysms. CONCLUSIONS: The clinical and postembolization outcomes in patients treated with the GDC system have improved in the past 5 years. Aneurysm recanalization, however, is still a major limitation of current GDC therapy. Follow-up angiography is mandatory after GDC embolization of cerebral aneurysms. Further technical and device improvements are mandatory to overcome current GDC limitations. PMID- 12744353 TI - Lack of improvement in cerebral metabolism after hyperoxia in severe head injury: a microdialysis study. AB - OBJECT: The authors investigated the effects of hyperoxia on brain tissue PO2 and on glucose metabolism in cerebral and adipose tissue after traumatic brain injury (TBI). METHODS: After 3 hours of ventilation with pure O2, 18 tests were performed on different days in eight comatose patients with TBI. Lactate, pyruvate, glucose, glutamate, and brain tissue PO2 were measured in the cerebral extracellular fluid (ECF) by using microdialysis. Analytes were also measured in the ECF of abdominal adipose tissue. After 3 hours of increase in the fraction of inspired O2, brain tissue PO2 rose from the baseline value of 32.7 +/- 18 to 122.6 +/- 45.2 mm Hg (p < 0.0001), whereas brain lactate dropped from its baseline (3.21 +/- 2.77 mmol/L), reaching its lowest value (2.90 +/- 2.58 mmol/L) after 3 hours of hyperoxia (p < 0.01). Pyruvate dropped as well, from 153 +/- 56 to 141 +/- 56 micromol/L (p < 0.05), so the lactate/pyruvate ratio did not change. No significant changes were observed in glucose and glutamate. The arteriovenous difference in O2 content dropped, although not significantly, from a baseline of 4.52 +/- 1.22 to 4.15 +/- 0.76 m/100 ml. The mean concentration of lactate in adipose tissue fell significantly as well (p < 0.01), but the lactate/pyruvate ratio did not change. CONCLUSIONS: Hyperoxia slightly reduced lactate levels in brain tissue after TBI. The estimated redox status of the cells, however, did not change and cerebral O2 extraction seemed to be reduced. These data indicate that oxidation of glucose was not improved by hyperoxia in cerebral and adipose tissue, and might even be impaired. PMID- 12744355 TI - Transsphenoidal microsurgical treatment of Cushing disease: postoperative assessment of surgical efficacy by application of an overnight low-dose dexamethasone suppression test. AB - OBJECT: Transsphenoidal adenomectomy with resection of a defined pituitary adenoma has been the treatment of choice for CD for the last 30 years. Surgical resection, however, may not always result in long-term remission of CD. This is particularly important in light of the high risk of morbidity and mortality in patients in the unsuccessfully treated cushingoid state. As such, it is interesting to identify prognostic factors that may predict the likelihood of long-term remission. METHODS: The authors review their series of 174 patients who have undergone transsphenoidal procedures for CD over a period of 20 years with minimum follow-up periods of 5 years. Selection of these patients was based on clinical, imaging, and laboratory criteria that included serum cortisol levels, loss of diurnal variation in serum cortisol levels, urinary free cortisol concentration, and results of a dexamethasone suppression test, petrosal sinus sampling, and corticotroph-releasing hormone stimulation tests as indicated. All patients who met the biochemical criteria underwent transsphenoidal microsurgery. The authors found an overall rate of remission of 74% at 5 years postoperatively. Patients in whom morning serum cortisol concentrations were lower than 3 microg/dl (83 nmol/L) on postoperative Day 3, following an overnight dexamethasone suppression test, had a 93% chance of remission at the 5-year follow-up examination. Patients with cortisol concentrations higher than this level uniformly failed to achieve long-term remission. CONCLUSIONS: Transsphenoidal microsurgery is an effective means of control for patients with adrenocorticotrophic hormone-producing microadenomas. Clinical outcome correlated well with the size of the tumor, as measured on preoperative imaging studies, and with postoperative morning cortisol levels following an overnight dexamethasone suppression test. Postoperative cortisol levels can be used as a useful prognostic indicator of the likelihood of future recurrence following transsphenoidal adenomectomy in CD. PMID- 12744356 TI - Anemia, testosterone, and pituitary adenoma in men. AB - OBJECT: Older men with clinically nonfunctioning pituitary tumors have been noted to be anemic, to have hypopituitarism, and to have low serum levels of testosterone. The authors hypothesized that men with pituitary adenomas and hypogonadism have a physiologically related decrease in hematocrit. METHODS: A retrospective analysis was conducted of 216 patients older than 50 years of age who harbored pituitary adenomas. In 100 men serum testosterone levels and a complete blood (cell) count (CBC) were obtained before treatment; a CBC was also acquired in a series of women with pituitary adenomas. Using clinical laboratory standards, anemia was defined as a hematocrit less than 40% in men and less than 35% in women. Thirty-one (46.3%) of 67 men with low serum concentrations of testosterone were anemic. In men with low levels of testosterone, the average hematocrit was 39.9%, compared with 45.6% for men with normal testosterone levels (p < 0.001). Men with macroadenomas were most likely to have both anemia and a low serum concentration of testosterone. Anemia was associated with a low level of testosterone, adjusting for tumor size (odds ratio 19, 95% confidence interval 4.86-77.03). Of patients with anemia, 84% were men and 16% were women (p < 0.001). The prevalence of anemia in women was low and was not correlated with tumor size. Men receiving testosterone replacement therapy had a significantly higher hematocrit value than men with low or normal testosterone levels. CONCLUSIONS: These findings support a direct relationship between serum testosterone levels and hematopoiesis in men, and demonstrate that hematopoiesis is compromised in men who have low concentrations of testosterone due to a pituitary adenoma. PMID- 12744357 TI - Systematic review of the prevention of delayed ischemic neurological deficits with hypertension, hypervolemia, and hemodilution therapy following subarachnoid hemorrhage. AB - OBJECT: There is uncertainty about the efficacy of hypertension, hypervolemia, and hemodilution (triple-H) therapy in reducing the occurrence of delayed ischemic neurological deficits (DINDs) and death after subarachnoid hemorrhage. The authors therefore conducted a systematic review to evaluate the efficacy of triple-H prevention in decreasing the rate of clinical vasospasm, DINDs, and death. METHODS: The authors systematically reviewed studies identified based on a MEDLINE, EMBASE, and COCHRANE Register search of articles published between 1966 and 2001, and reference lists of identified articles. An independent assessment of each study's methodological quality, population, intervention, and outcomes (rates of symptomatic vasospasm, DINDs, and death) was performed. Summary relative risk estimates were calculated for the main outcomes using fixed- or random-effect models, as appropriate. Only four prospective, comparative studies with a total of 488 patients were identified. The median internal validity score was 0.5 (range 0-2); the median external validity score was 3 (range 2-6). Compared with no prevention, triple-H therapy was associated with a reduced risk of symptomatic vasospasm (relative risk [RR] 0.45, 95% confidence interval [CI] 0.32-0.65), but not DIND (RR 0.54, 95% CI 0.2-1.49). The risk of death was higher (RR 0.68, 95% CI 0.53-0.87). Sensitivity analyses including only randomized, controlled trials showed no evidence of statistically significant results for these major end points. CONCLUSIONS: The paucity of information and important limitations in the design of the studies analyzed preclude evaluation of the efficacy of triple-H prevention and formulation of any recommendations regarding its use for the prevention of cerebral vasospasm. PMID- 12744358 TI - Edema after intracerebral hemorrhage: correlations with coagulation parameters and treatment. AB - OBJECT: Development of edema is known to contribute to poor outcome after spontaneous intracerebral hemorrhage (ICH). Recent research has identified thrombin as a key mediator in the development of edema in animal models; however, little has been published correlating the coagulation cascade and edema in humans. METHODS: In this retrospective clinical study of 80 patients with spontaneous supratentorial ICH, the authors sought to identify factors associated with edema development and outcome, including lesion imaging parameters, anticoagulant use, international normalized ratio and platelet count on hospital admission, and treatment with mannitol and steroid medications. A multivariate model was used to identify edema volume, use of mannitol, elevated blood glucose, and the presence of intraventricular hemorrhage as predictors of poor outcome at the time patients were discharged from the hospital. The authors developed a quadratic model for predicting edema volume against time by using a random coefficients model, and found that edema peaks between Days 5 and 6 after onset of ICH. The volume of the hemorrhage and the platelet count correlated significantly with edema volume within the first 24 hours post-ICH in the multiple regression analysis (p < 0.0001, r2 = 0.75). Edema growth during the first 5 days post-ICH also correlated with the platelet count, with an increasing platelet count associated with an increasing growth of edema (p = 0.0013). CONCLUSIONS: The authors propose that factors released from activated platelets at the site of hemorrhage, for example vascular endothelial growth factor, may interact with thrombin to increase vascular permeability and contribute to the development of edema. PMID- 12744359 TI - Surgical outcomes of 654 ulnar nerve lesions. AB - OBJECT: In this article the authors present a retrospective analysis of 654 surgical outcomes in patients with ulnar nerve entrapments, injuries, and tumors during a 30-year period. METHODS: Data were gathered between 1968 and 1998 at Louisiana State University Health Sciences Center. Mechanisms of injuries or lesions included 460 entrapments at the elbow level (70%), 76 lacerations (12%), 52 stretches/contusions (8%), 34 fractures/dislocations (5%), 12 gunshot wounds (2%), two injection-induced injuries (0.3%), and 13 nerve sheath tumors (2%). In cases of entrapment, direct operative recordings uniformly demonstrated a slowing of conduction at the elbow, even in cases in which preoperative noninvasive studies had been nondiagnostic. Intraoperative electrical "inching" studies also demonstrated significant conduction abnormalities that lie just proximal to and through the olecranon notch rather than distal, beneath the flexor carpi ulnaris muscle. There were only eight exceptions to this. Lesions not in continuity due to the injury required primary or secondary end-to-end sutures or graft repair. Aided by intraoperative nerve action potential recording, lesions in continuity received either external or internal neurolysis and split repair or resection followed by end-to-end suture or graft repair. Functional recoveries of Grade 3 or better were seen in 81 (92%) of 88 patients who underwent neurolysis, 42 (72%) of 58 patients who received suture repair, and 24 (67%) of 36 patients who received graft repair. Nevertheless, fewer Grade 4 or 5 recoveries were reached than those seen in patients with radial or median nerve injuries. Nerve sheath tumors were resected with preservation of preoperative function in five of seven patients. CONCLUSIONS: Although difficult to obtain, useful functional recovery can be achieved with proper surgical management of ulnar nerve entrapments and injuries. PMID- 12744360 TI - Outcomes of surgery in 1019 brachial plexus lesions treated at Louisiana State University Health Sciences Center. AB - OBJECT: Outcomes of 1019 brachial plexus lesions in patients who underwent surgery at Louisiana State University Health Sciences Center during a 30-year period are reviewed in this paper to provide management guidelines. METHODS: Causes of brachial plexus lesions included 509 stretches/contusions (50%), 161 plexus tumors (16%), 160 thoracic outlet syndromes (TOSs, 16%), 118 gunshot wounds (12%), and 71 lacerations (7%). Many features of clinical presentation, including prior treatment, patient's neurological status, results of electrophysiological studies, intraoperative findings, and postoperative level of function, were studied. The minimum follow-up period was 18 months and the mean follow-up period was 42 months. Repairs were best for injuries located at the C 5, C-6, and C-7 levels, the upper and middle trunk, the lateral cord to the musculocutaneous nerve, and the median and posterior cords to the axillary and radial nerves. Conversely, results were poor for injuries at the C-8 and T-1 levels, and for lower trunk and medial cord lesions, with the exception of injuries of the medial cord to the median nerve. Outcomes were most favorable when patients were carefully evaluated and selected for surgery, although variables such as lesion type, location, and severity, as well as time since injury also affected outcome. This was true also of TOSs and tumors arising from the plexus, especially if they had not been surgically treated previously. CONCLUSIONS: Surgical exploration and repair of brachial plexus lesions is technically feasible and favorable outcomes can be achieved if patients are thoroughly evaluated and appropriately selected. PMID- 12744361 TI - Complications of invasive subdural grid monitoring in children with epilepsy. AB - OBJECT: This study was performed to evaluate the complications of invasive subdural grid monitoring during epilepsy surgery in children. METHODS: The authors retrospectively reviewed the records of 35 consecutive children with intractable localization-related epilepsy who underwent invasive video electroencephalography (EEG) with subdural grid electrodes at The Hospital for Sick Children between 1996 and 2001. After subdural grid monitoring and identification of the epileptic regions, cortical excisions and/or multiple subpial transections (MSTs) were performed. Complications after these procedures were then categorized as either surgical or neurological. There were 17 male and 18 female patients whose mean age was 11.7 years. The duration of epilepsy before surgery ranged from 2 to 17 years (mean 8.3 years). Fifteen children (43%) had previously undergone surgical procedures for epilepsy. The number of electrodes on the grids ranged from 40 to 117 (mean 95). During invasive video EEG, cerebrospinal fluid leaks occurred in seven patients. Also, cerebral edema (five patients), subdural hematoma (five patients), and intracerebral hematoma (three patients) were observed on postprocedural imaging studies but did not require surgical intervention. Hypertrophic scars on the scalp were observed in nine patients. There were three infections, including one case of osteomyelitis and two superficial wound infections. Blood loss and the amounts of subsequent transfusions correlated directly with the size and number of electrodes on the grids (p < 0.001). Twenty-eight children derived significant benefit from cortical resections and MSTs, with a more than 50% reduction of seizures and a mean follow-up period of 30 months. CONCLUSIONS: The results of this study indicate that carefully selected pediatric patients with intractable epilepsy can benefit from subdural invasive monitoring procedures that entail definite but acceptable risks. PMID- 12744362 TI - Changes in cerebrospinal fluid hydrodynamics following endoscopic third ventriculostomy for shunt-dependent noncommunicating hydrocephalus. AB - OBJECT: The aim of this study was to analyze physiological changes in cerebrospinal fluid (CSF) dynamics following endoscopic third ventriculostomy (ETV) for shunt-dependent noncommunicating hydrocephalus. METHODS: . Clinical data obtained in 15 patients treated with ETV for shunt malfunction were analyzed. Magnetic resonance imaging studies demonstrated the obstruction of the ventricular system preoperatively. After ETV, the existing shunt system was removed and a continuous extraventricular drain, set at 30 cm H2O in height, was installed to measure daily amounts of CSF outflow. Cerebrospinal fluid dynamics after ETV were also evaluated using 111In-diethylenetriamine pentaacetic acid radioisotope cisternography in six of 15 patients within 1 month of the procedure. Three patients underwent cisternography at 6 months after ETV. Cisternograms were obtained at 1, 5, 24, and 48 hours after injection of the radioisotope. To study CSF absorptive capacity, ratios of radioisotope counts at 48 and 5 hours after injection were calculated (C48:C5). Seven of 15 patients had daily outflows of CSF of less than 20 ml; this volume decreased quickly within a few days. The other eight patients demonstrated an outflow of more than 150 ml of CSF for several days, three of whom had signs of transiently increased intracranial pressure. Their CSF outflow volume decreased gradually and symptoms improved within 1 week. Ratios of C48:C5 were within normal limits in five of six patients who had undergone cisternography 1 month after ETV. These ratios were decreased in all three patientswho had undergone cisternography at 6 months after ETV compared with that measured at 1 month after the procedure. CONCLUSIONS: Our data suggest that CSF dynamics convert from a shunt-dependent state to a shunt independent state within I week following ETV in patients with shunt-dependent noncommunicating hydrocephalus. Nonetheless, intraventricular pressure does not decrease quickly in certain cases. Cerebrospinal fluid absorptive capacity or CSF circulation through the subarachnoid space may show further improvement several months after ETV. PMID- 12744363 TI - Endoscopic third ventriculostomy for shunt dysfunction in occlusive hydrocephalus: long-term follow up and review. AB - OBJECT: Endoscopic third ventriculostomy (ETV) is the treatment of choice for occlusive (noncommunicating) hydrocephalus. Nevertheless, its routine use in patients who have previously undergone shunt placement is still not generally accepted. The authors' aim was to investigate the long-term effects of ETV in a group of prospectively chosen patients. METHODS: Patients who underwent ETV and had previously undergone shunt placement for occlusive hydrocephalus were followed prospectively for at least 3 years (range 36-103 months, mean 63.6 months). Nine female and eight male patients ranging from 8 to 54 years of age (mean 32 years) had undergone shunt placement 0.7 to 23.5 years (mean 8.1 years) before ETV. Fifteen patients were admitted with underdrainage and two with overdrainage. In six cases, ETV was performed as an emergency operation. The origin of hydrocephalus was aqueductal stenosis in 12 cases and aqueductal compression by a tumor in two cases. Three patients suffered from a fourth ventricle outlet syndrome, and in two patients an additional malresorptive component was suspected. Thirteen patients underwent ETV with shunt removal and insertion of an external drain in one session. The drain served as a safety measure; it could be opened if raised intracranial pressure or ventricular dilation was observed on postoperative imaging studies. In the other four patients the shunt was initially ligated and then removed during a second operation. Fourteen patients (82%) have remained shunt free. The other three patients, including the two with an additional malresorptive component, needed shunt reimplantation 3 days, 2 weeks, or 7 months after ETV. CONCLUSIONS: Use of ETV is safe and effective for the treatment for shunt dysfunction in patients with obstructive hydrocephalus. PMID- 12744364 TI - Treatment of staphylococcal ventriculitis associated with external cerebrospinal fluid drains: a prospective randomized trial of intravenous compared with intraventricular vancomycin therapy. AB - OBJECT: Staphylococcal ventriculitis may be a complication in temporary external ventricular drains (EVDs). The limited penetration of vancomycin into the cerebrospinal fluid (CSF) is well known; the pharmacodynamics and efficacy of systemically compared with intraventricularly administered vancomycin is examined in this prospective study. METHODS: Ten patients in whom EVDs were implanted to treat intracranial hemorrhage and who were suffering from drain-associated ventriculitis were randomized into two treatment groups. Five of these patients (median age 47 years) were treated with 2 g/day vancomycin administered intravenously (four infusions/day, Group 1), and the other five(median age 49 years) received 10 mg vancomycin intraventricularly once daily (Group 2). Vancomycin levels were measured in serum and CSF six times a day. The maximum vancomycin level in CSF was 1.73 +/- 0.4 micro/ml in Group 1 and 565.58 +/- 168.71 microg/ml 1 hour after vancomycin application in Group 2 (mean +/- standard deviation). Vancomycin levels above the recommended trough level of 5 microg/ml in CSF were never reached in Group 1, whereas in Group 2 they below the trough level (3.74 +/- 0.66 microg/ml) only at 21 hours after intraventricular vancomycin application. The vancomycin level in the serum was constant within therapeutic levels in Group 1, whereas in Group 2 in most instances vancomycin was almost below a measurable concentration. In both groups bacteriologically and laboratory-confirmed CSF clearance could be obtained. CONCLUSIONS: Intraventricular vancomycin application is a safe and efficacious treatment modality in drain-associated ventriculitis, with much higher vancomycin levels being achieved in the ventricular CSF than by intravenous administration. PMID- 12744365 TI - Pseudotumor cerebri syndrome: venous sinus obstruction and its treatment with stent placement. AB - OBJECT: Pseudotumor cerebri, or benign intracranial hypertension, is a condition of raised intracranial pressure in the absence of a mass lesion or cerebral edema. It is characterized by headache and visual deterioration that may culminate in blindness. Pseudotumor cerebri is caused by venous sinus obstruction in an unknown percentage of cases. The purpose of this study was to investigate the role of cerebral venous sinus disease in pseudotumor cerebri and the potential of endoluminal venous sinus stent placement as a new treatment. METHODS: Nine consecutive patients in whom diagnoses of pseudotumor cerebri had been made underwent examination with direct retrograde cerebral venography (DRCV) and manometry to characterize the morphological features and venous pressures in their cerebral venous sinuses. The cerebrospinal fluid (CSF) pressure was measured simultaneously in two patients. If patients had an amenable lesion they were treated using an endoluminal venous sinus stent. Five patients demonstrated morphological obstruction of the venous transverse sinuses (TSs). All lesions were associated with a distinct pressure gradient and raised proximal venous sinus pressures. Four patients underwent stent insertion in the venous sinuses and reported that their headaches improved immediately after the procedure and remained so at 6 months. Vision was improved in three patients, whereas it remained poor in one despite normalized CSF pressures. CONCLUSIONS: Patients with pseudotumor cerebri should be evaluated with DRCV and manometry because venous TS obstruction is probably more common than is currently appreciated. In patients with a lesion of the venous sinuses, treatment with an endoluminal venous sinus stent is a viable alternative for amenable lesions. PMID- 12744366 TI - Methionine positron emission tomography of recurrent metastatic brain tumor and radiation necrosis after stereotactic radiosurgery: is a differential diagnosis possible? AB - OBJECT: In this study the authors examined how to differentiate radiation necrosis from recurrent metastatic brain tumor following stereotactic radiosurgery by using positron emission tomography (PET) with L-[methyl 11C]methionine (MET). METHODS: In 21 adult patients with suspected recurrent metastatic brain tumor or radiation injury, MET-PET scans were obtained. These patients had previously undergone stereotactic radiosurgery and subsequent contrast-enhanced magnetic resonance (MR) examinations before nuclear medicine imaging. Positron emission tomography images were obtained as a static scan of 10 minutes performed 20 minutes after injection of 370 MBq of MET. On MET-PET scans, the portion of the tumor with the highest accumulation of MET was selected as the region of interest (ROI), and the ratio of tumor tissue to normal tissue (T/N) was defined as the mean counts of radioisotope per pixel in the tumor divided by the mean counts per pixel in normal gray matter. The standardized uptake value (SUV) was calculated using the same ROI in the tumor. The accuracy of the MET-PET scan was evaluated by correlating findings with results of subsequent histological analysis (11 cases) or, in cases in which surgery or biopsy was not performed, with subsequent clinical course and MR imaging findings (10 cases). Histological examinations performed in 11 cases showed viable tumor cells with necrosis in nine and necrosis with no viable tumor cells in two. Another 10 cases were characterized as radiation necrosis because the patients exhibited stable neurological symptoms with no sign of massive enlargement of the lesion on follow up MR images after 5 months. The mean T/N was 1.15 in the radiation necrosis group (12 cases) and 1.62 in the tumor recurrence group (nine cases). The mean SUV was 1.78 in the necrosis group and 2.5 in the recurrence group. There were statistically significant differences between the recurrence and necrosis groups in T/N and SUV. Furthermore, the borderline T/N value was 1,42 according to a 2 x 2 factorial table (high T/N or low T/N, recurrence or necrosis). From this result, the sensitivity and specificity of MET-PET scanning in detecting tumor recurrence were determined to be 77.8 and 100%, respectively. CONCLUSIONS: The use of MET-PET scanning is a sensitive and accurate technique for differentiating between metastatic brain tumor recurrence and radiation necrosis following stereotactic radiosurgery. This study reveals important information for creating strategies to treat postradiation reactions. PMID- 12744367 TI - Trauma-induced tumorigenesis of cells implanted into the rat spinal cord. AB - OBJECT: Findings in several clinical cases have suggested a correlation between tumor formation and previous injury to the central nervous system (CNS); however, the relationship between trauma and tumorigenesis has not been investigated well experimentally. In this study the authors provide evidence correlating tumorigenesis with trauma in the rat spinal cord. METHODS: A glial cell line, C6R G/H, which expresses green fluorescent protein (GFP) and hygromycin phosphotransferase (HPT), was implanted into normal and injured rat spinal cords. In all rats in which the cells were implanted into an injured site, locomotor function deteriorated and histological analysis demonstrated glioblastoma multiforme by 6 weeks; tumorigenesis was correlated with a loss of both GFP expression and resistance to hygromycin treatment. In contrast, no evidence of tumor formation was found at 6 weeks in rats in which the cells were implanted into healthy tissue. When C6R-G/H cells were treated with contused spinal cord extract in culture before implantation, they lost GFP expression and hygromycin resistance, and later formed tumors after implantation into normal spinal cord. CONCLUSIONS: The findings of this study indicate that trauma can induce tumorigenesis. Implantation of C6R-G/H cells into traumatized spinal cords resulted in their transformation, which was signaled by loss of GFP expression and hygromycin resistance accompanied by tumor formation. Exposure to extracts derived from injured spinal cord produced similar transformation and gene expression changes, as well as tumor formation after such cells were implanted into normal cords. Care, therefore, should be taken when cells are implanted into an injured CNS because of potential mutagenesis due to trauma-induced factors. PMID- 12744368 TI - Amyloid beta accumulation in axons after traumatic brain injury in humans. AB - OBJECT: Although plaques composed of amyloid beta (AD) have been found shortly after traumatic brain injury (TBI) in humans, the source for this Abeta has not been identified. In the present study, the authors explored the potential relationship between Abeta accumulation in damaged axons and associated Abeta plaque formation. METHODS: The authors performed an immunohistochemical analysis of paraffin-embedded sections of brain from 12 patients who died after TBI and from two control patients by using antibodies selective for Abeta peptides, amyloid precursor protein (APP), and neurofilament (NF) proteins. In nine brain injured patients, extensive colocalizations of Abeta, APP, and NF protein were found in swollen axons. Many of these immunoreactive axonal profiles were present close to Abeta plaques or were surrounded by Abeta staining, which spread out into the tissue. Immunoreactive profiles were not found in the brains of the control patients. CONCLUSIONS: The results of this study indicate that damaged axons can serve as a large reservoir of Abeta, which may contribute to Abeta plaque formation after TBI in humans. PMID- 12744369 TI - Glial cell line-derived neurotrophic factor-supplemented hibernation of fetal ventral mesencephalic neurons for transplantation in Parkinson disease: long-term storage. AB - OBJECT: Transplantation of fetal dopaminergic tissue is being investigated in animal models and clinical trials for its potential as a treatment for advanced Parkinson disease. At the same time, the availability of fetal tissue is limited, making its storage time prior to transplantation a key practical issue. Although it results in a smaller percentage of surviving cells. a longer storage time enables fetal tissue obtained over several days to be pooled for transplantation in a recipient. Glial cell line-derived neurotrophic factor (GDNF) has been shown to improve survival of human dopaminergic tissue that has been stored prior to transplantation. The objective of this study was to evaluate the effects on fetal dopaminergic tissue of GDNF-supplemented hibernation for extended periods of 6 to 15 days. METHODS: The ventral mesencephalon (VM) was harvested in a total of 27 14-day-old rat fetuses, and three VMs were cultured immediately (fresh control group). The remaining 24 VMs were divided sagittally along the midline to yield 48 equal pieces of hemimesencephalon. Twenty-four pieces were stored with GDNF supplemented hibernation medium for 6, 9, 12, or 15 days, and the 24 "partner" hemimesencephalon pieces were stored in control hibernation medium for the same periods of time. Tissue was cultured for 48 hours and processed for tyrosine hydroxylase (TH) immunoreactivity and double-stained with cresyl violet. Cell counts for all cultures and the percentage of TH-immunoreactive cells were obtained. The percentage of TH-immunoreactive cells for the fresh control group was 6.3 +/- 0.5%. The percentage of TH-immunoreactive cells in cultures derived from tissue stored in GDNF-supplemented medium was significantly increased at 6 and 9 days posthibernation compared with the fresh control group and the "partner" groups stored in hibernation medium only. No significant increase in the percentage of TH-immunoreactive cells was observed in the 12- and 15-day groups. CONCLUSIONS: In this study the authors have demonstrated that fetal dopaminergic tissue can be safely stored for up to 9 days in GDNF-supplemented hibernation medium. Furthermore, the percentage of TH-immunoreactive cells is significantly increased after 6 and 9 days of storage in this medium, improving the yield of TH-immunoreactive cells prior to transplantation. These observations have practical clinical implications for collecting fetal dopaminergic cells and improving their survival after transplantation. PMID- 12744370 TI - Polysialylated neural cell adhesion molecules expressed in human pituitary tumors and related to extrasellar invasion. AB - OBJECT: Pituitary adenomas are usually benign tumors; however, some behave aggressively and metastasize. Until now, no specific marker of aggressive behavior or malignancy has been found. The polysialylated neural cell adhesion molecule (NCAM), which is highly expressed in embryonic tissues such as the brain and pituitary, is detected in some and neuroendocrine tumors. Because polysialylation has been implicated in the regulation of cell growth and migration, polysialylated NCAM expression has been considered as a prognostic marker in such tumors. METHODS: In the present study, the authors analyzed polysialylated NCAM expression in 82 pituitary tumors from humans: 49 secreting adenomas, 32 nonfunctioning adenomas, and one growth hormone and prolactin secreting carcinoma associated with acromegaly and spinal and liver metastases. Based on immunohistochemical analyses, the tumors were classified as somatotropic (22 tumors), prolactinoma (14 tumors), corticotropic (17 tumors), and gonadotropic or so-called null cell adenomas (28 tumors). Assessment of polysialylated NCAM was performed using three different methods (immunohistochemical analysis, Western blot analysis, and enzyme-linked immunosorbent assay) with a specific mouse monoclonal immunoglobulin M (Men B) that recognizes polysialic acid on NCAM. Tumoral NCAM expression was also evaluated with the aid of immunohistochemical analysis. Using this method, NCAM and polysialylated NCAM were studied in six healthy pituitaries. In addition, correlations were investigated using three statistical methods (chi-square test, nonparametric Mann-Whitney U-test, and principal component analysis) to compare tumoral polysialylated NCAM expression and seven parameters (tumor size and type, intrasphenoidal or cavernous sinus invasion, Ki-67 index, mitoses, and patient age and sex). Neural cell adhesion molecules were expressed in the healthy anterior pituitary and in all tumors. In contrast, polysialylated NCAM was not found in the healthy pituitary gland, but was expressed in 46.3% of typical pituitary tumors and 85% of the tumors selected as highly aggressive, including one carcinoma and three tumors with histological characteristics that raised suspicion of malignancy. There was no significant correlation between polysialylated NCAM expression and tumor size, tumor type, Ki-67 index, mitoses, or patient age and sex. In contrast, the expression of polysialylated NCAM, which was sensitive to endoneuraminidase-N treatment, was strongly correlated with tumor invasion (p < 0.0001). CONCLUSIONS: In pituitary tumors in humans, expression of polysialylated NCAM is strongly related to tumor invasion and confirms the clinical diagnosis of aggressiveness. PMID- 12744371 TI - Functional motor neurons differentiating from mouse multipotent spinal cord precursor cells in culture and after transplantation into transected sciatic nerve. AB - OBJECT: One of the current challenges in neurobiology is to ensure that neural precursor cells differentiate into specific neuron types, so that they can be used for transplantation purposes in patients with neuron loss. The goal of this study was to determine if spinal cord precursor cells could differentiate into motor neurons both in culture and following transplantation into a transected sciatic nerve. METHODS: In cultures with trophic factors, neurons differentiate from embryonic precursor cells and express motor neuronal markers such as choline acetyltransferase (ChAT), Islet-1, and REG2. Reverse transcription-polymerase chain reaction analysis has also demonstrated the expression of Islet-1 in differentiated cultures. A coculture preparation of neurospheres and skeletal myocytes was used to show the formation of neuromuscular connections between precursor cell-derived neurons and myocytes both immunohistochemically and electrophysiologically. Following various survival intervals, precursor cells transplanted distal to a transection of the sciatic nerve differentiated into neurons expressing the motor neuron markers ChAT and the alpha1 1.2 (class C, L type) voltage-sensitive Ca++ channel subunit. These cells extended axons into the muscle, where they formed cholinergic terminals. CONCLUSIONS: These results demonstrate that motor neurons can differentiate from spinal cord neural precursor cells grown in culture as well as following transplantation into a transected peripheral nerve. PMID- 12744372 TI - Treatment of patients with intractable obsessive-compulsive disorder with anterior capsular stimulation. Case report. AB - Obsessive-compulsive disorder (OCD) is a common, chronic, disabling anxiety disorder characterized by recurrent obsessive thoughts and uncontrolled repetitive acts. Although many patients respond to various pharmacological treatments, there is a cohort of patients with intractable or refractory disease. The authors present the case of a patient with intractable OCD who was treated with bilateral electrical stimulators, which were stereotactically placed in the anterior limbs of the internal capsules. Following psychiatric consultation and 10 years of empirical medication regimens for OCD, a woman was referred for neurosurgical evaluation. After informed consent had been obtained from the patient, the authors placed bilateral stimulator leads in the anterior limbs of the internal capsules. The stereotactic coordinates were based on data in pertinent current literature. The stimulation parameters, which are presented in this paper, were set at 2 weeks and reviewed at 6 weeks and 3 months postoperatively. No changes were required. Postoperative analysis included evaluation by the patient's referring psychiatrist, a second independent psychiatrist, and pre- and postoperative administration of the Yale-Brown Obsessive Compulsive Scale. A marked improvement was noted in thispatient's OCD symptomatology and general psychosocial function. Previous documentation of patient responses to psychosurgical procedures for intractable or refractory OCD has been met with little enthusiasm, presumably because of the invasiveness and irreversibility of the surgery. In this report the authors suggest that deep brain stimulation of appropriate targets may be an effective and safe treatment for certain patients with OCD and a potentially reversible treatment for those patients who do not obtain therapeutic benefit. PMID- 12744373 TI - Amusia following resection of a Heschl gyrus glioma. Case report. AB - The incidence and character of neurological deficits following resection of glial neoplasms localized to the Heschl gyrus are currently unknown. In this series, the authors report the clinical presentation, management, and postoperative course of three patients with right hemisphere Heschl gyrus gliomas, one of whom developed difficulty with music production and comprehension postoperatively. Resection of right hemisphere Heschl gyms gliomas can result in deficits involving music comprehension. Preliminary evidence suggests that when these deficits occur, they may be transient in nature. PMID- 12744375 TI - Intracranial stent placement for the treatment of a carotid-cavernous fistula associated with intracranial angioplasty. Case report. AB - The authors report a case of an iatrogenic carotid-cavernous fistula (CCF) associated with intracranial angioplasty. Angioplasty was performed using a 3 x 10-mm Open Sail coronary balloon in a patient with high-grade stenosis of the left cavernous internal carotid artery (ICA). After angioplasty, a perforation developed in the cavernous ICA, resulting in a CCF. A 3.5 x 9-mm S670 coronary stent was used to treat the fistula. To the authors' knowledge, this is the first reported case in which a CCF developed after angioplasty was performed using a coronary balloon. Long-term angiographic and clinical evaluation is needed to test the suitability and durability of intracranial angioplasty and stent placement in the treatment of symptomatic intracranial stenosis. PMID- 12744374 TI - Rapid development of Chiari I malformation in an infant with Seckel syndrome and craniosynostosis. Case report and review of the literature. AB - To illustrate the rapidity with which a child can develop a severe, symptomatic Chiari I malformation, the authors present the case of a 3-month-old infant with Seckel syndrome (microcephaly, micrognathia, craniosynostosis, and multiple other abnormalities) and posterior sagittal and bilateral lambdoid synostosis. The infant underwent magnetic resonance (MR) imaging shortly after birth; the initial image demonstrated the cerebellar tonsils in the posterior fossa, with no herniation. He subsequently developed severe apneic episodes and bradycardia; repeated MR imaging at 3 months demonstrated severe tonsillar herniation with compression of the brainstem. The child underwent posterior fossa remodeling surgery, including release of the posterior sagittal and lambdoid sutures and decompression of the Chiari I malformation. The patient's apnea gradually improved; however, he died of complications of pneumonia and sepsis several weeks later. The authors identified from the literature 21 patients in whom there was a documented MR image or other neuroimage that did not reveal evidence of a Chiari I malformation, followed by a subsequent study with clear documentation of the presence of Chiari I malformation. The interval between the initial study and the development of the tonsillar herniation ranged from 11 days to 18.5 years. In most cases, a lumbar cerebrospinal fluid (CSF) diversion had been performed. This patient developed a severely symptomatic Chiari I malformation during a 3-month period. These reports illustrate that the Chiari I malformation can develop rapidly in the face of increased intracranial pressure, craniosynostosis, and spinal CSF diversion. PMID- 12744376 TI - Synchronous subarachnoid drop metastases from a pituitary adenoma with multiple recurrences. Case report. AB - The authors report the case of a 49-year-old man with synchronous drop metastases from a multiply recurrent somatotroph pituitary adenoma. The metastatic lesions were found in the subarachnoid space of the cauda equina and foramen magnum 18 years after the initial diagnosis of the disease. Five transsphenoidal resections had previously failed to cure the sellar tumor. Two of these, performed 4 and 5 years before the patient's current presentation, had been complicated by cerebrospinal fluid rhinorrhea that necessitated lumbar drainage. Resections of the two subarachnoid lesions, separated by 14 months, removed pathologically aggressive pituitary adenomas. There were no signs of local recurrence or subarachnoid dissemination of disease during the postoperative follow-up periods, which lasted 18 and 4 months, respectively. Previous cases of subarachnoid spread of a pituitary adenoma have been associated with multiple intracranial metastases, multiple intraspinal metastases, or widely disseminated disease. This case demonstrates that subarachnoid metastasis of a pituitary adenoma, particularly when it follows multiple operations, is not invariably widely disseminated or associated with a very poor prognosis. PMID- 12744377 TI - A syndrome of spontaneous cerebral and cervical artery dissections with angiolipomatosis. Report of two cases. AB - A primary or systemic arteriopathy is frequently suspected in patients with spontaneous cerebral or cervical artery dissections. The authors report on two patients with such dissections accompanied by angiolipomatosis, a previously unreported association, and propose a common developmental defect in these patients. A 50-year-old man with subcutaneous angiolipomatosis developed painful monocular blindness. Angiography studies revealed a spontaneous extracranial internal carotid artery (ICA) dissection and an ipsilateral fusiform intracranial ICA aneurysm. The ICA dissection was treated with aspirin, and after 6 months a craniotomy was performed. The aneurysm was found to be fusiform; it involved the entire supraclinoid portion of the ICA, and was wrapped with cotton. A 49-year old man with a congenitally bicuspid aortic valve and subcutaneous angiolipomatosis developed posterior neck pain. Magnetic resonance imaging and angiography demonstrated a fusiform distal vertebral artery aneurysm. A craniotomy was performed and the aneurysm was found to incorporate the posterior inferior cerebellar artery as well as a perforating artery: the lesion was wrapped cotton. The tunica media of the arteries of the head and neck as well as the aortic valvular cusps are derived from neural crest cells, and angiolipomatosis has been associated with tumors of neural crest derivation. These associations indicate that a neural crest disorder may be the underlying abnormality in these patients. PMID- 12744378 TI - Vascularized temporalis muscle flap for the treatment of otorrhea. Technical note. AB - The surgical treatment for cerebral spinal fluid (CSF) fistulas provides closure of the bone and dural defects and prevents the recurrence of brain herniation and CSF fistula. The two main approaches used are the transmastoid and middle fossa ones. The authors review the results of performing a modified middle fossa approach with a vascularized temporalis muscle flap to create a barrier between the repaired dural and bone defects. Fifteen consecutive cases of CSF fistulas treated at the authors' institution were retrospectively reviewed. All patients presented with otorrhea. Eleven patients had previously undergone ear surgery. A middle fossa approach was followed in all cases. The authors used a thin but watertight and vascularly preserved temporalis muscle flap that had been dissected from the medial side of the temporalis muscle and was laid intracranially on the floor of the middle fossa, between the repaired dura mater and petrous bone. The median follow-up period was 2.5 years. None of the patients experienced recurrence of otorrhea or meningitis. There was no complication related to the intracranial temporalis muscle flap (for example, seizures or increased intracranial pressure caused by muscle swelling). One patient developed hydrocephalus, which resolved after the placement of a ventriculoperitoneal shunt 2 months later. The thin, vascularized muscle flap created an excellent barrier against the recurrence of CSF fistulas and also avoided the risk of increased intracranial pressure caused by muscle swelling. This technique is particularly useful in refractory cases. PMID- 12744379 TI - Clival osteoblastoma in a child. Case illustration. PMID- 12744380 TI - Remarkable regression of optic glioma in an infant. Case illustration. PMID- 12744381 TI - Transcutaneous coil, stent, and balloon migration following endovascular treatment of a cervical carotid artery aneurysm. Case illustration. PMID- 12744382 TI - Third ventriculostomy for internal hydrocephalus complicated by unrecognized subdural hygroma and hematoma: a case report of a patient treated by Dr. Walter Dandy. AB - The authors review the case of a patient treated by Dr. Walter Dandy. When the patient was a young child he underwent two right transtemporal third ventriculostomies during which he sustained an unrecognized contralateral subdural hygroma and a chronic subdural hematoma with a mild infantile hemiparesis. He was able to complete high school, albeit at a slower pace than usual. As an adult he held several limited employment positions, lived at home for several decades, and was later cared for at a nursing home for a short time. The patient died when he was 66 years of age. PMID- 12744383 TI - Distal PICA aneurysms. PMID- 12744384 TI - Cerebrospinal fluid volume. PMID- 12744385 TI - Prevention of early rebleeding. PMID- 12744386 TI - Prevention of early rebleeding. PMID- 12744388 TI - Prevention of early rebleeding. PMID- 12744387 TI - Prevention of early rebleeding. PMID- 12744389 TI - Spine patient outcomes research trial. PMID- 12744390 TI - Experimental study of noise properties of a Ti:sapphire femtosecond laser. AB - The fidelity of a coherent link between optical and microwave frequencies is largely determined by noise processes in a mode-locked femtosecond laser. This work presents an experimental study of the noise properties of a Ti:sapphire femtosecond laser. It includes measurements of pulse repetition rate fluctuations and shot noise exhibited by the Ti:sapphire femtosecond laser. Based on the results of noise measurements, the fractional frequency stability of a microwave signal produced by the femtosecond laser has been evaluated. PMID- 12744391 TI - Piezoelectric ultrasonic micromotor with 1.5 mm diameter. AB - A piezoelectric ultrasonic micromotor has been developed using a lead zirconate titanate (PZT) ceramic/metal composite tube stator that was 1.5 mm in diameter and 7 mm in length. The micromotor was operated in its first bending vibration mode (approximately 70 kHz), producing speeds from hundreds to over 2000 rpm in both rotational directions. The maximum torque-output was 45 microN-m, which is far superior to previous PZT thin film-based micromotors. This micromotor showed good reliability and stability for more than 300 hours of continued operation. PMID- 12744392 TI - Finite element simulation for a new disc-type ultrasonic stator. AB - This paper is concerned with the development of a new disc-type piezoelectric ultrasonic stator. Linear piezoelectric, mechanical, and piezoelectro-mechanic behaviors of a metal disc structure embedded with piezoelectric actuator are considered. Using a finite element method, a dynamic formulary is modeled for the new disc-type piezoelectric ultrasonic stator. In this model, a 3-dimensional (D) mechanical element with an extra electrical degree of freedom is used to simulate dynamic vibration modes and analyze characteristic responses such as electrical impedance response, phase response, and mechanical frequency response for a new disc-type piezoelectric ultrasonic stator. An adaptive boundary condition, simple support condition with three nonequal-triangular fixed points near the edge for the mechanism design of a new disc-type piezoelectric ultrasonic stator is defined so that a lateral elliptical motion of the contact point between stator and rotor can be realized for driving the rotor. The finite element results have been compared with the experimental measurements. As a result, the analysis model seems to be similar to the real condition. PMID- 12744393 TI - Nanometer stepping drives of surface acoustic wave motor. AB - High resolution (from nanometer to subnanometer) stepping drives of a surface acoustic wave motor are presented. It was shown that step displacement was easily controlled by adjusting a number of driving waves, using a steel ball slider equipped with permanent magnet for preload. By means of this open loop control, the step displacement was controlled from centimeter-order to submicrometer order. In this paper, using a silicon slider equipped with a ball bearing linear guide, the stepping motions of a surface acoustic wave motor were investigated. A laser interferometer equipped with a 2-picometer resolution displacement demodulator was introduced. Motions of the slider ranging from several hundreds of nanometers to several nanometers in each step displacement were observed. Reduction of the driving waves down to 25 cycles, under a 100 Vpeak driving voltage and a 30 N preload condition, generated about 2 nm stepping motion using our experimental setup under an open loop condition. We also demonstrated subnanometer step movements. These experimental results indicated that the surface acoustic wave motor has an ability of subnanometer positioning with a centimeter-level stroke. PMID- 12744394 TI - A precise measurement of some nonlinear effects and its application to the evaluation of nonlinear elastic constants of quartz and GaPO4. AB - This paper deals with a precise measurement of amplitude frequency and intermodulation effects, and its application to the evaluation of nonlinear elastic constants of quartz and gallium orthophosphate (GaPO4). An evaluation is based on the methods used previously concerning determination of the higher-order material constants in the quartz. Using a measurement of the intermodulation products and measurement of drive level dependence of resonant frequency of quartz resonators, we have determined some effective elastic constants of fourth order. The computer-based method of solution of the set of equations gives an access to obtain a number of effective nonlinear stiffnesses of fourth order. The measurements and computer solutions are performed on different Y-cuts resonators, both for quartz and gallium orthophosphate resonators, vibrating in fundamental thickness shear mode. The experimental results are discussed. PMID- 12744395 TI - A comparison of the performance of time-delay estimators in medical ultrasound. AB - Time-delay estimation (TDE) is a common operation in ultrasound signal processing. In applications such as blood flow estimation, elastography, phase aberration correction, and many more, the quality of final results is heavily dependent upon the performance of the time-delay estimator implemented. In the past years, several algorithms have been developed and applied in medical ultrasound, sonar, radar, and other fields. In this paper we analyze the performances of the widely used normalized and non-normalized correlations, along with normalized covariance, sum absolute differences (SAD), sum squared differences (SSD), hybrid-sign correlation, polarity-coincidence correlation, and the Meyr-Spies method. These techniques have been applied to simulated ultrasound radio frequency (RF) data under a variety of conditions. We show how parameters, which include center frequency, fractional bandwidth, kernel window size, signal decorrelation, and signal-to-noise ratio (SNR) affect the quality of the delay estimate. Simulation results also are compared with a theoretical performance limit set by the Cramer-Rao lower bound (CRLB). Results show that, for high SNR, high signal correlation, and large kernel size, all of the algorithms closely match the theoretical bound, with relative performances that vary by as much as 20%. As conditions degrade, the performances of various algorithms differ more significantly. For signals with a correlation level of 0.98, SNR of 30 dB, center frequency of 5 MHz with a fractional bandwidth of 0.5, and kernel size of 2 micros, the standard deviation of the jitter error is on the order of few nanoseconds. Normalized correlation, normalized covariance, and SSD have an approximately equal jitter error of 2.23 ns (the value predicted by the CRLB is 2.073 ns), whereas the polarity-coincidence correlation performs less well with a jitter error of 2.74 ns. PMID- 12744396 TI - Characterization of trabecular bone using the backscattered spectral centroid shift. AB - Ultrasonic attenuation in bone in vivo is generally measured using a through transmission method at the calcaneus. Although attenuation in calcaneus has been demonstrated to be a useful predictor for osteoporotic fracture risk, measurements at other clinically important sites, such as hip and spine, could potentially contain additional useful diagnostic information. Through transmission measurements may not be feasible at these sites due to complex bone shapes and the increased amount of intervening soft tissue. Centroid shift from the backscattered signal is an index of attenuation slope and has been used previously to characterize soft tissues. In this paper, centroid shift from signals backscattered from 30 trabecular bone samples in vitro were measured. Attenuation slope also was measured using a through-transmission method. The correlation coefficient between centroid shift and attenuation slope was -0.71. The 95% confidence interval was (-0.86, -0.47). These results suggest that the backscattered spectral centroid shift may contain useful diagnostic information potentially applicable to hip and spine. PMID- 12744397 TI - Optimization of attenuation estimation in reflection for in vivo human dermis characterization at 20 MHz. AB - In vivo skin attenuation estimators must be applicable to backscattered radio frequency signals obtained in a pulse-echo configuration. This work compares three such estimators: short-time Fourier multinarrowband (MNB), short-time Fourier centroid shift (FC), and autoregressive centroid shift (ARC). All provide estimations of the attenuation slope (beta, dB x cm(-1) x MHz(-1)); MNB also provides an independent estimation of the mean attenuation level (IA, dB x cm( 1)). Practical approaches are proposed for data windowing, spectral variance characterization, and bandwidth selection. Then, based on simulated data, FC and ARC were selected as the best (compromise between bias and variance) attenuation slope estimators. The FC, ARC, and MNB were applied to in vivo human skin data acquired at 20 MHz to estimate betaFC, betaARC, and IA(MNB), respectively (without diffraction correction, between 11 and 27 MHz). Lateral heterogeneity had less effect and day-to-day reproducibility was smaller for IA than for beta. The IA and betaARC were dependent on pressure applied to skin during acquisition and IA on room and skin-surface temperatures. Negative values of IA imply that IA and beta may be influenced not only by skin's attenuation but also by structural heterogeneity across dermal depth. Even so, IA was correlated to subject age and IA, betaFC, and betaARC were dependent on subject gender. Thus, in vivo attenuation measurements reveal interesting variations with subject age and gender and thus appeared promising to detect skin structure modifications. PMID- 12744398 TI - A rapid signal processing technique to remove the effect of dispersion from guided wave signals. AB - Guided acoustic and ultrasonic waves have been utilized in various manners for non-destructive evaluation and testing. If a guided wave mode is dispersive, a pulse of energy will spread out in space and time as it propagates. For a long range guided wave inspection application, this constrains the choice of operating point to regions on the dispersion curves where dispersion effects are small. A signal processing technique is presented that enables this constraint on operating point to be relaxed. The technique makes use of a priori knowledge of the dispersion characteristics of a guided wave mode to map signals from the time to distance domains. In the mapping process, dispersed signals are compressed to their original shape. The theoretical basis of the technique is described and an efficient numerical implementation is presented. The robustness of the technique to inaccuracies in the dispersion data is also addressed. The application of the technique to experimental data is shown and the resulting improvement in spatial resolution is demonstrated. The implications of using dispersion compensation in practical systems are briefly discussed. PMID- 12744400 TI - Simulation of Lamb wave propagation for the characterization of complex structures. AB - Reliable numerical simulation techniques represent a very valuable tool for analysis. For this purpose we investigated the applicability of the local interaction simulation approach (LISA) to the study of the propagation of Lamb waves in complex structures. The LISA allows very fast and flexible simulations, especially in conjunction with parallel processing, and it is particularly useful for complex (heterogeneous, anisotropic, attenuative, and/or nonlinear) media. We present simulations performed on a glass fiber reinforced plate, initially undamaged and then with a hole passing through its thickness (passing-by hole). In order to give a validation of the method, the results are compared with experimental data. Then we analyze the interaction of Lamb waves with notches, delaminations, and complex structures. In the first case the discontinuity due to a notch generates mode conversion, which may be used to predict the defect shape and size. In the case of a single delamination, the most striking "signature" is a time-shift delay, which may be observed in the temporal evolution of the signal recorded by a receiver. We also present some results obtained on a geometrically complex structure. Due to the inherent discontinuities, a wealth of propagation mechanisms are observed, which can be exploited for the purpose of quantitative nondestructive evaluation (NDE). PMID- 12744399 TI - Experimental spatial sampling study of the real-time ultrasonic pulse-echo BAI mode imaging technique. AB - The ultrasonic pulse-echo backscattered amplitude integral (BAI)-mode imaging technique has been developed to inspect the seal integrity of hermetically sealed, flexible food packages. With a focused 17.3-MHz transducer acquiring radio frequency (RF) echo data in a static rectilinear stop-and-go pattern, this technique was able to reliably detect channel defects as small as 38 microm in diameter and occasionally detect 6-microm-diameter channels. This contribution presents our experimental spatial sampling study of the BAI-mode imaging technique with a continuous zigzag scanning protocol that simulates a real-time production line inspection method in continuous motion. Two transducers (f/2 17.3 MHz and f/3 20.3 MHz) were used to acquire RF echo data in a zigzag raster pattern from plastic film samples bearing rectilinear point reflector arrays of varying grid spacings. The average BAI-value difference (deltaBAI) between defective and intact regions and the contrast-to-noise ratio (CNR) were used to assess image quality as a function of three spatial sampling variables: transducer spatial scanning step size, array sample grid spacing, and transducer 6-dB pulse-echo focal beam spot size. For a given grid size, the deltaBAI and CNR degraded as scanning step size in each spatial dimension increased. There is an engineering trade-off between the BAI-mode image quality and the transducer spatial sampling. The optimal spatial sampling step size has been identified to be between one and two times the -6-dB pulse-echo focal beam lateral diameter. PMID- 12744401 TI - Calculation and measurement of electromechanical coupling coefficient of capacitive micromachined ultrasonic transducers. AB - The electromechanical coupling coefficient is an important figure of merit of ultrasonic transducers. The transducer bandwidth is determined by the electromechanical coupling efficiency. The coupling coefficient is, by definition, the ratio of delivered mechanical energy to the stored total energy in the transducer. In this paper, we present the calculation and measurement of coupling coefficient for capacitive micromachined ultrasonic transducers (CMUTs). The finite element method (FEM) is used for our calculations, and the FEM results are compared with the analytical results obtained with parallel plate approximation. The effect of series and parallel capacitances in the CMUT also is investigated. The FEM calculations of the CMUT indicate that the electromechanical coupling coefficient is independent of any series capacitance that may exist in the structure. The series capacitance, however, alters the collapse voltage of the membrane. The parallel parasitic capacitance that may exist in a CMUT or is external to the transducer reduces the coupling coefficient at a given bias voltage. At the collapse, regardless of the parasitics, the coupling coefficient reaches unity. Our experimental measurements confirm a coupling coefficient of 0.85 before collapse, and measurements are in agreement with theory. PMID- 12744402 TI - Tissue stiffness imaging method using temporal variation of ultrasound speckle pattern. AB - Applying vibration to a medium makes it vibrate. The resulting change in scatterer distribution inside the medium due to applied vibration changes the speckle pattern of ultrasound images. In this case, scatterers in a hard medium experience small displacements, and those in a soft medium experience large displacements. As a result, the amount of speckle pattern brightness change in ultrasound images is related to the tissue stiffness. Using this dependency, a two-dimensional profile of relative tissue stiffness can be constructed qualitatively at the display pixel resolution by determining at each pixel the standard deviation and/or the difference between minimum and maximum values over a certain number of consecutive B-mode images. Experiments with phantoms show that the softer the tissue, the larger the standard deviation. The proposed imaging modality is a simple yet practical method of resolving hard cysts surrounded by soft background in a phantom using B-mode frame data only. PMID- 12744403 TI - Dynamic response analysis of pyroelectric sensitive element for thermal imaging. AB - Temperature distributions under periodic thermal excitations and the responsivity of a pyroelectric device consisting of a cover layer, infrared absorber, metal contact, sensitive pyroelectric element, interconnecting column, and bulk silicon are found. Some results of numerical thermal modeling and analysis of exact expressions for a few extreme cases are presented. Pyroelectric responses of real structures are compared with the response of a single pyroelectric element in air as a limiting case of maximum sensitivity. The analytical approximations and numerical simulation show that the frequency response of the multilayered structure consists of different parts with simple frequency dependencies. In the region of high frequencies of light modulation, the responsivity is proportional to omega(-1), at low frequencies approximately omega(-0.5), and, in the region of intermediate frequencies, the voltage responsivity is independent of frequency. PMID- 12744404 TI - Removal of bacterial endotoxin from root surface with Er:YAG laser. AB - PURPOSE: To determine the potential of 2.94 microm Er:YAG laser radiation to remove bacterial endotoxin from root surfaces. MATERIALS AND METHODS: 40 extracted teeth were divided into four groups of 10 samples each. A 16 mm2 area of the root surface on each sample was inoculated with an aliquot of 7 microl of a lipopolysaccharide suspension at a concentration of 50 IU/ml. LPS was derived from a non-oral Escherichia coli strain (E. coli 055:B5). Source of laser radiation was an Er:YAG laser emitting pulsed infrared radiation at a wavelength of 2.94 microm, with a pulse duration of 250 micros, and a pulse repetition rate of 15 pps. Three specimen groups were irradiated with 105 laser pulses at a radiation energy of 60 mJ, 100 mJ and 140 mJ. One specimen group was untreated (control). The LPS concentration with each sample was determined using a chromogenic, quantitative Limulus-amoebocyte-lysate assay. Statistical analysis was ANOVA and Scheffe-test. RESULTS: Mean LPS yield from the untreated control samples was 50.1 (+/- 35.9) IU/ml. Following laser irradiation the average LPS on the root surfaces was 19.86 (+/- 14.4) IU/ml at 60 mJ, 12.86 (+/- 8.1) IU/ml at 100 mJ and 8.58 (+/- 4.9) IU/ml at 140 mJ. PMID- 12744405 TI - Water treeing--a potential mechanism for degradation of dentin adhesives. AB - PURPOSE: To test the hypothesis that water channels are present along the adhesive-dentin interface that can be detected using transmission electron microscopy examination of tracer penetration. MATERIALS AND METHODS: Different total-etch adhesives (Single Bond, Excite, and PQ1) and self-etch adhesives (Prompt L-Pop, Reactmer Bond, and an experimental self-etch adhesive) were bonded to dentin and enamel. Bonded resin-tooth slabs were immersed in 50 wt% conventional silver nitrate or 50 wt% basic, ammoniacal silver nitrate for 24 hours, exposed to a photodeveloping solution, and prepared for transmission electron microscopy. RESULTS: With both types of silver nitrate, all adhesives exhibited nanoleakage within hybrid layers. In addition, water trees in the form of interconnecting, dendritic silver deposits were seen along the surface of the hybrid layers that extended perpendicularly into the adhesive layers. With ammoniacal silver nitrate, additional isolated, unconnected silver grains were observed within the adhesives. PMID- 12744407 TI - Fatigue behavior of the resin-resin bond of partially replaced resin-based composite restorations. AB - PURPOSES: (1) To evaluate different pre-treatment modes for partial resin-based composite repair using a shear bond strength fatigue design on aged specimens, and (2) to investigate the effect of these pre-treatments on aged and partially replaced Class II resin-based composite restorations. MATERIALS AND METHODS: (Study 1) 150 composite discs (Tetric Ceram) were light-cured, secondary cured, and stored in water for 365 days. The specimens were randomly assigned to five pretreatment groups (n=30): (1) Roughening with a finishing diamond bur + dentin bonding system (DBS, Syntac Classic), (2) Roughening with a silicon carbide bur + DBS, (3) Roughening with a silicon carbide bur + DBS + Tetric Flow, (4) silica coating (CoJet) + DBS, and (5) silica-coating + dentin bonding agent + Tetric Flow. The disks were restored with fresh repair composite cylinders and after 24 hour water storage, initial bond strengths (n=10) and fatigue bond strengths (n=20; 5,000 cycles in shear mode) were evaluated and fracture modes determined under a SEM. (Study 2): 40 Class II restorations (EBS Multi/Pertac II) were placed in extracted human third molars. The specimens were stored for 365 days and then replaced partially by removing the proximal box and leaving the occlusal part. The partial repairs (Pertac II) were placed after identical pre-treatment modes A-E (n=8). After thermomechanical loading (100,000 x 50 N, simultaneously x2,500 at 5 degrees C/55 degrees C), marginal quality between old and fresh composite was evaluated by SEM at x200 magnification and dye penetration was observed by light microscopy under x50 magnification. RESULTS: Initial repair bond strengths were significantly higher for the groups with additional application of flowable composite compared to composite only (P<0.05; Mann Whitney U test). Diamond finishing revealed the lowest bond strengths. Fatigue bond strengths exhibited the worst results for Group 1 as well, but were similar for Groups 2-5. In the Class II partial repair simulation, Groups 3 and 5 with additional flowable lining exhibited significantly better marginal quality and significantly lower leakage, whereas the other groups revealed up to 100% separation between aged and freshly applied composite. PMID- 12744406 TI - Fracture strength of cusp replacing resin composite restorations. AB - PURPOSE: To assess the influence of an additional shoulder preparation on the fracture strength of a cusp-replacing direct resin composite restoration in a premolar that previously had an amalgam MOD restoration followed by fracture of a cusp. MATERIALS AND METHODS: Two preparation designs were tested. In extracted sound premolars an MOD amalgam cavity was simulated followed by fracture of a cusp (Group A). Group B was the same as Group A but a shoulder was added along the cervical outline of the preparation. As a control group, cusp fracture was simulated but no MOD amalgam cavities were prepared. For each group, 14 preparations were made using a copy milling technique. All 42 preparations were restored using a direct resin composite restoration technique. Restorations were loaded until fracture. RESULTS: The fracture strength of the preparation with shoulder was not significantly different from the preparation without the shoulder. The fracture strength of the preparations in the reference group, which had no resistance form, was significantly less than in Groups A and B (ANOVA, P<0.05). PMID- 12744408 TI - A 3-year clinical evaluation of glass-ionomer cements used as fissure sealants. AB - PURPOSE: To evaluate the retention and caries-preventive effectiveness of two ionomeric materials (conventional and resin-modified), used as fissure sealants. MATERIALS AND METHODS: 100 children (6-8 years old) with a total of 400 permanent first molars received 200 conventional glass-ionomer (Ketac Bond) and 200 resin modified glass-ionomer (Vitremer) sealants. Additionally, 108 children constituted the control group (432 teeth). Two dentists assisted by dental hygienists performed the sealant application. Clinical evaluations were carried out 6, 12, 24, and 36 months after the sealant application by two other dentists, not carrying out clinical procedures, previously calibrated (Kappa > 0.75). RESULTS: Total retention rates of 26%, 12%, 3%, and 4% for Ketac Bond and 61%, 31%, 14%, and 13% for Vitremer, being 6, 12, 24, and 36 months after clinical evaluation, respectively. The differences between the two materials were statistically significant. The experimental groups showed a caries incidence of 93%, 78%, 49%, and 56% lower than the control group (P< 0.01) for the four evaluation periods, respectively. In conclusion, the retention rates of ionomeric materials were low. Nevertheless, these materials showed a cariostatic effect, supported by statistically lower caries incidence in experimental groups compared to control group. Presence of active incipient caries was statistically associated with caries incidence in the first molars after 36 months, in relation to either experimental or control group. PMID- 12744409 TI - Resin-dentin interfaces of endodontically-treated restored teeth. AB - PURPOSE: To compare by confocal and scanning electron microscopy the post-resin dentin interfaces of endodontically-treated teeth which had been restored with dentin-bonding systems and composite and needed to be extracted after 6 months to 6 years of clinical service. MATERIALS AND METHODS: Sixteen single rooted teeth showing proximal lesions penetrating the pulp chamber were included in the investigation and divided into two groups. Group 1 teeth were endodontically treated, restored with carbon fiber posts and resin-based composite. Group 2 teeth were treated in the same way and covered with porcelain fused-to-metal crowns. RESULTS: Two thirds of the observed interfaces showed resin penetration of the root dentin in the middle and coronal third of the post space preparation, while only one third of the interface showed resin infiltration in the apical third of the post space preparation. Debonding of the adhesive from the resin infiltrated dentin area and debonding of the composite cement from the adhesive were the most frequently observed failure modes and were observed in one third of the observed interfaces. There was no difference between the post-resin-dentin interfaces of teeth covered and teeth not covered with porcelain-fused to metal crowns. Teeth extracted after longer periods of clinical service showed more abundant areas of debonding of the adhesive from the resin-dentin interdiffusion zone. The presence of bacteria in resin infiltrated dentin tubules was observed in two specimens. PMID- 12744410 TI - 3-year evaluation of a new open sandwich technique in Class II cavities. AB - PURPOSE: To evaluate the durability of a new open sandwich restoration with, as shown in a previous scanning electron microscope study, improved interfacial adaptation. A polyacid-modified resin-based composite (PMRC; compomer) was placed as an intermediate layer and covered with resin composite (RC). A direct RC restoration was used as control. MATERIALS AND METHODS: Of 57 patients, each received at least one pair of Class II restorations, one PMRC/RC open sandwich and one RC control. In total, 75 pairs of Class II restorations, 68 premolars and 82 molars, all in occlusion, were placed by two dentists. Most of the cavities were surrounded by enamel. The restorations were evaluated at baseline, 6, 12, 24 and 36 months by a slightly modified USPHS criteria. RESULTS: Five of 148 restorations evaluated during 3 years were rated as unacceptable. Two, one in each group, because of endodontic treatment and one RC restoration was replaced because of tooth fracture. Secondary caries was observed contiguous to one restoration in each group at the 36-month recall. Except for the two patients with pulpitis, none of the others reported postoperative sensitivity. No significant differences were seen between the restoration techniques. For marginal adaptation a significant change occurred between baseline and 6 months in both groups. For marginal discoloration, a significant change was observed at 6 months in the sandwich group and at 36 months in the RC group. Color match of the resin composite material changed significantly in both groups at 36 months. It was concluded that both techniques showed good durability during the 3-year follow up. PMID- 12744411 TI - Efficacy of self-etching primer on sealing margins of Class II restorations. AB - PURPOSE: To evaluate sealing ability of different types of restorative-adhesive combinations and to correlate etch patterns with leakage scores. MATERIALS AND METHODS: 56 molars were selected and divided randomly in four groups of 14 specimens each. A standardized adhesive Class II preparation with the cervical margin placed 1 mm below the CEJ and an occlusal reduction of 2 mm was performed. No bevels were utilized in the preparation. Four combinations of bonding system/restorative material were tested. Group 1: Excite (EX) in combination with Tetric Ceram (TC) as control; Group 2: Prompt-L-Pop (PP1) applied for 15 seconds in combination with TC; Group 3: Etch and Prime 3.0 in combination with Definite restorative material (EP); Group 4: Prompt-L-Pop (PP2) applied for 30 seconds in combination with TC. The bonding systems (Groups 1, 2 and 3) and all restorative materials were used following strictly manufacturers' instructions. The resin composite was applied following an incremental technique. Ten specimens of each group were processed for leakage test. The specimens were sectioned with a diamond saw in three different areas in mesial-distal direction. Two different operators evaluated the sections blindly for scoring leakage at cervical and occlusal margins. The highest score for the sections of each tooth area was selected for scoring and further statistical analysis. The results of the staining measurements were statistically evaluated using the Kruskal-Wallis non parametric analysis of variance with Bonferroni alpha protection. The level of statistical significance was defined as P<0.05. The remaining four specimens of each group were kept in a 37% HCl solution for 48 hours to dissolve the dental structures and to observe the resin replica of the cavities by SEM. RESULTS: EX showed less dye penetration at occlusal margins than the other three groups, while no statistically significant differences were found at the dentin margin. The SEM observations showed rougher and more uniform enamel etch pattern when phosphoric acid (EX) was applied than that obtained with self-etching adhesive systems. Resin tags and adhesive lateral branches were noted in all groups at the dentin site. PMID- 12744412 TI - Effects of a carbamide peroxide agent and desensitizing dentifrices on enamel microhardness. AB - PURPOSE: To evaluate the in vitro microhardness of enamel treated with a 10% carbamide peroxide agent and two desensitizing dentifrices at different bleaching times. MATERIALS AND METHODS: A 10% carbamide peroxide bleaching agent was evaluated (Rembrandt 10%) (REM). A placebo agent was used as a control group (PLA). The bleaching and the placebo agents were applied to human enamel dental fragments for 8 hours per day, followed by immersion for 5 minutes in a slurry solution of desensitizing dentifrices: Sensodyne (S) or Sensodyne Fluor (SF). During the remaining time, the enamel fragments were individually stored in 13.5 ml of artificial saliva. Knoop microhardness measurements were performed at baseline, 8 hours, 7, 14, 21, 28, 35 and 42 days of treatment and at 7 and 14 days of a post-treatment period. RESULTS: Analysis of variance and Tukey's test showed no differences in enamel microhardness for REM + SF (P=0.069) and PLA + SF (P=0.93) within each time interval. The dental fragments treated with REM + S and PLA + S showed an increase in microhardness values within each time interval (P<0.0001). There were significant differences among the treatment agents from the 28th to the 56th day. The use of 10% carbamide peroxide bleaching and a desensitizing dentifrice significantly increased the enamel microhardness values during the bleaching treatment and after 14 days after the completion of the treatment. After the post-treatment period, the enamel fragments treated with a placebo agent and with a 10% carbamide peroxide agent and with a desensitizing fluoride dentifrice maintained the baseline values. PMID- 12744413 TI - Resin adhesion to carious dentin. AB - PURPOSE: To investigate the microtensile bond strength (microTBS) of a self etching priming adhesive system to normal, caries-affected and caries-infected dentin, and to observe the ultrastructure of the resin-dentin interface by transmission electron microscopy (TEM). MATERIALS AND METHODS: Twelve extracted human molar teeth with deep occlusal caries were stained with caries detector solution and ground flat occlusally. The red-stained soft dentin was classified as caries-infected. The surrounding discolored dentin was classified as caries affected dentin. The surrounding normal dentin served as a control. The entire flat surface was bonded with Clearfil Liner Bond 2V (CV) and covered with resin composite to form a composite crown 5 mm high. One day later the specimens were serially sectioned vertically into multiple slabs 0.8 mm thick. Under microscopic observation, the specimens were divided into normal or caries-infected or caries affected dentin. These regions were isolated by cutting away the remaining dentin to form hour-glass shapes with the smallest surface area at the test site. After measuring the areas, the specimens were fixed to a microtensile tester and pulled under tension to failure. Additional slabs that were not used for bond strength tests were processed for TEM. Bond strength data were analyzed by Kruskal-Wallis one-way ANOVA and Student-Newman-Keuls multiple comparisons. RESULTS: The microTBS of CV to normal, caries-affected and caries-infected dentin were 45 +/- 10 MPa, 30 +/- 10 MPa, 10 +/- 5 MPa, respectively. TEM images showed that CV formed thin hybrid layers that were less than 1 microm thick in normal dentin, but that were between 6-8 microm thick in caries-affected dentin. Bacteria were only sparsely observed in the dentin tubules of bonded caries-affected dentin. However, in caries-infected dentin, an unusual interface was seen in which carious bacteria within disorganized non-banded collagen fibrils could be seen embedded by the adhesive. The hybrid layer in caries-infected dentin was found to be 30-60 microm thick. PMID- 12744414 TI - Effects of grit blasting and silanization on bond strengths of a resin luting cement to Belleglass HP indirect composite. AB - PURPOSE: To investigate the extent of bonding possible of a resin luting cement with three opacities of Belleglass HP, representing different filler type and loading levels in a highly cross-linked resin matrix. MATERIALS AND METHODS: Belleglass HP (3 opacities: opaque dentin, translucent dentin, cuspal enamel) was prepared as solid notched discs (Group A) or luted halves to create notched discs (Groups B-F). Luted samples were split through the notch on the Instron for conditions: as-is after molding (B), grit blasted with 50 microm Al2O3 (C), grit blasted and silanated (D) grit blasted, silanated, and stored 1 week (E), and grit blasted and treated with Artglass liquid (F). All the samples were stored in distilled water at 37 degrees C for 24 hours before testing. The fractured surfaces were examined by light and scanning electron microscopy (SEM) to assess the effect of the surface treatments. RESULTS: The mean fracture resistance (+/- S.D) of the tested groups ranged from 7.6 (+/- 1.9 N mm(-1)) for Group A to 13.9 (+/- 5.4 N mm(-1)) for Group D. Two-way analysis of variance showed no significant interaction (P>0.05) between product opacities & surface treatments. The bonded inlay test halves in Groups B and C failed adhesively between the inlay test halves and the luting cement whereas the other three test groups (D,E,F) failed cohesively in the luting cement. One-way analysis of variance with post-hoc paired group Tukey testing revealed that Group D (13.9 +/- 5.4 N mm(-1)) performed significantly better (P<0.05) than Groups A (7.6 +/- 1.9 N mm(-1), B (9.2 +/- 3.7 N mm(-1)) or C (7.8 +/- 1.6 N mm(-1)). No statistical difference was found between Groups D to F (P>0.05). PMID- 12744415 TI - Formation of Streptococcus mutans biofilm following toothbrushing with regular and whitening toothpastes. AB - PURPOSE: To examine the effect of toothbrushing with different brands of regular (Meridol, Colgate) and whitening toothpaste (Pearl Weiss, Rembrant, Email Diamant) on bacterial adhesion onto enamel surfaces in vitro. MATERIALS AND METHODS: Extracted intact human teeth were brushed twice a day, for 3 minutes with the tested toothpaste for a period of 7 days. Sucrose dependent adhesion of Streptococcus mutans to the tested teeth was measured using pre-radioactive labeled bacteria. RESULTS: S. mutans adhered more to enamel surfaces, pre-brushed with the tested toothpastes, than to teeth which were not brushed. Different profiles of bacterial adhesion were recorded. Both Diamant and Rembrant toothpastes increased bacterial affinity toward the enamel surface as compared to the other toothpastes. Meridol and Pearl Weiss demonstrated a similar adhesion effect as the etching control while brushing with Colgate demonstrated the least bacterial adhesion. PMID- 12744416 TI - Direct pulp capping with an auto-cured sealant resin and a self-etching primer. AB - PURPOSE: To evaluate an auto-cured resin as a direct pulp capping material on exposed pulp with or without a self-etching primer. MATERIALS AND METHODS: Class V cavities were prepared on the facial surfaces of 90 intact monkey teeth, and the pulps were intentionally exposed with a carbide bur through the cavity floor. Each exposed pulp was capped with a commercially available adhesive resin system (Clearfil Liner Bond 2V: Group LB) or an auto-cured sealant resin (Teethmate-S) with and without the use of a self-etching primer (Group TMP and Group TM, respectively). The cavities were restored with an adhesive resin and a hybrid resin-based composite (Clearfil AP-X). Inflammatory cell infiltration and dentin bridge formation of the exposed pulp as well as protrusion of the exposed pulp tissue into the cavities were evaluated histologically at 3, 30 and 90 days. RESULTS: Slight inflammatory cell infiltration was the main inflammatory reaction of the exposed pulp, and the exposed area became occluded with dentin bridges as the observation period increased. However, significantly higher incidence of slight inflammatory cell infiltration was found in Group TM than in Group LB at 30 days (P<0.05). Group TM showed significantly higher incidence of protrusion of pulp tissue than Groups LB and TMP through all tested periods (P<0.05). PMID- 12744417 TI - Bioactive molecules and the future of pulp therapy. AB - Direct capping of bioactive molecules or implantation of these molecules in the pulp may induce the formation of reparative dentin and coronal or radicular pulp mineralization. In this review, we summarize what is known and/or assumed on the biological mechanisms of these therapies. We report on the effects which were obtained experimentally in rat maxillary molars by implantation of Bone Sialoprotein (BSP)/collagen pellets and Specific Amelogenin Gene Splice Products [A+4] and [A-4]) adsorbed on agarose beads. The effects were compared with those of BMP-7 (OP-1) and Ca(OH)2. Depending on the molecule that was used, we obtained either the formation of a dentin bridge, or the filling of the pulp in the mesial part of the coronal pulp chamber, or the total mineralization of the root canal. These molecules may provide new therapeutic tools in the next future in dentistry. PMID- 12744418 TI - Effects of physician gender on patient satisfaction. AB - OBJECTIVES: To measure the impact of physician gender on patient satisfaction, controlling for confounding patient variables, and to examine the extent to which differences in satisfaction with male and female physicians can be explained by physician practice styles. METHOD: New adult patients (n=509) were randomized to see male and female primary care physicians at a university medical center outpatient facility. Patient sociodemographics and self-reported health status (using the Medical Outcomes Study Short Form-36) were measured before the initial visit, and satisfaction with the physician was measured immediately following the visit. The entire medical encounter was videotaped and physician practice style was later analyzed using the Davis Observation Code. RESULTS: Female physicians spent a significantly greater proportion of the visit on preventive services and counseling than male physicians did, and male physicians devoted more time to technical practice behaviors and discussions of substance abuse. Visit length was not significantly different for male and female physicians. Patients of female physicians were more satisfied than were those of male physicians, even after adjusting for patient characteristics, visit length, and physician practice style behaviors. CONCLUSION: Patient satisfaction with primary care physicians appears to be influenced not only by patient characteristics and physician behaviors, but also by the gender of the provider. Possible explanations for this may be that psychosocial aspects of the physician-patient interaction are different for male and female physicians. Patients may also bring expectations about female physicians to the encounter, presuming them to be more empathetic, nurturing, and responsive. PMID- 12744419 TI - What happens when health care providers ask about intimate partner violence? A description of consequences from the perspectives of female survivors. AB - OBJECTIVE: To describe positive and negative consequences of health care screening for intimate partner violence from the perspectives of female survivors. METHOD: We conducted 7 semistructured focus group interviews with 41 women in battered women's shelters or intimate partner violence support groups. RESULTS: Positive consequences of screening included: recognizing that the violence was a problem, decreased isolation, and feeling that the medical provider cared. Negative consequences included: feeling judged by the provider, increased anxiety about the unknown, feeling that the intervention protocol was cumbersome or intrusive, and disappointment in the provider's response. CONCLUSION: We found that both positive and negative consequences can result from screening for intimate partner violence and that they are related to provider behavior. The positive consequences described by the participants reflect changes in their attitudes, thoughts, and feelings that may precede help seeking. A better understanding of consequences can help providers tailor screening approaches and interventions for intimate partner violence. PMID- 12744420 TI - A population-based assessment of chronic unexplained vulvar pain: have we underestimated the prevalence of vulvodynia? AB - OBJECTIVES: To estimate the prevalence of unexplained chronic vulvar pain (burning or sharp knife like pain or pain on contact) in an ethnically diverse population-based sample of women. METHODS: We used town census directories to identify 4915 women age 18 to 64 from 5 ethnically diverse Boston communities and asked them to complete a self-administered questionnaire pertaining to current and past chronic lower genital tract discomfort (response rate 68%). We calculated the cumulative incidence and 95% confidence intervals by demographic and reproductive characteristics. Approximately 16% of respondents reported histories of chronic burning, knife like pain, or pain on contact that lasted for at least 3 months or longer, and nearly 7% were experiencing the problem at the time of the survey. Chronic vulvar pain on contact decreased with increasing age, but the cumulative incidence of chronic burning and knife like pain was similar across all ages. Contrary to earlier clinical assessments, white and African American women reported similar lifetime prevalences. However, Hispanic women were 80% more likely to experience chronic vulvar pain than were white and African American women. Women with histories of chronic vulvar pain were 7 to 8 times more likely to report difficulty and great pain with their first tampon use than were women without such histories. Nearly 40% of women chose not to seek treatment, and of those who did, 60% saw 3 or more doctors, many of whom could not provide a diagnosis. CONCLUSION: Chronic unexplained vulvar pain is a highly prevalent disorder that is often misdiagnosed. PMID- 12744421 TI - Biology and mental illness: a historical perspective. AB - A century ago, psychiatrists understood women's susceptibility to mental illness in terms of their unique biology. Although contemporary physicians certainly do not share late 19th-century psychiatrists' biases about women and the social order, the similarities between today's emphasis on women's biology and earlier explanations of the relationship between women's biology and mental illness bear investigation. This paper reviews the history of medical ideas about the connection between women's reproductive organs and their mental health and questions modern assumptions about that connection. PMID- 12744422 TI - Condom use errors and problems among young women who put condoms on their male partners. AB - OBJECTIVES: To comprehensively assess condom use errors and problems reported by women who apply condoms, given the lack of data on this aspect of condom use. METHODS: Data from a questionnaire survey, using a 3-month recall period, was analyzed for a convenience sample of 102 college women who put condoms on their male partner(s) for sex (vaginal, anal, or oral). The percentage of the sample reporting that an error or problem occurred at least once is presented. RESULTS: Fifty-one percent put the condom on after starting sex, and 15% took the condom off before ending sex. Forty-eight percent wanted a condom but didn't have one, 46% did not leave space at the tip, 30% put the condom on wrong side up and had to flip the condom over, 26% did not use a lubricant, and 15% wanted a water based lubricant and did not have any available. Twenty-five percent reported that their partners lost erections in association with condom use. Twenty-eight percent reported breakage, slippage, or both. Higher error scores were found for those who reported: 1) breakage or slippage, 2) partner erection problems, 3) use of another form of contraception, and 4) not having received instruction in correct condom use. CONCLUSION: Educating women as well as men about correct condom use, in addition to promoting consistent condom use, may be an important public health strategy. PMID- 12744423 TI - Postmenopausal osteoporosis risk management in primary care: how well does it adhere to national practice guidelines? AB - OBJECTIVE: The National Osteoporosis Foundation (NOF) practice guideline provides explicit recommendations for postmenopausal osteoporosis risk assessment and management. This study evaluated primary care adherence to the recommendations and factors associated with such adherence. METHOD: Postmenopausal women 40 years old and older were sampled in the primary care clinic of a US tertiary hospital. We received 469 completed questionnaires that measured the respondent's demographics and osteoporosis risk factors; current and historical osteoporosis counseling, testing, and treatment; and whether or not she had a primary care clinician. Based on information provided, we determined whether each woman was receiving care that followed the NOF guideline. Univariate and multivariate analyses identified factors associated with guideline adherence. RESULTS: Mean age was 69 years; 53% were white and 38% black; 15% had prior fractures. Using NOF criteria, 19.8 % were at moderate risk and 80.2 % were at high risk for osteoporosis. Overall, 57.6% (95% confidence interval 53.1, 62.1) of patients received risk management in accordance with the NOF guideline. The adherence rate was higher in the high-risk group (60.9% v 44.1%, p=.003), in those who had primary care clinicians (62.0% v 43.5%, p=.006), and in nonblack patients (white v black 67.1% v 43.0%, p<.001; other v black 65.0% v 43.0%, p=.012). These associations remained statistically significant after multivariate adjustments. Adherence was not associated with the sex of the primary care clinician. CONCLUSIONS: More than half of postmenopausal women in our study were receiving osteoporosis risk management that followed the NOF practice guideline. Those who were at higher risk, who were not black, or who had primary care clinicians were more likely to receive care in accordance with the NOF guideline, regardless of the sex of the clinician. PMID- 12744424 TI - Cardiovascular risk factors among women in Mississippi in the 1990s. AB - OBJECTIVES: To identify the prevalence of cardiovascular disease risk factors among Mississippi women from 1991 to 1999, using data from the Behavior Risk Factor Surveillance Survey (BRFSS) and to examine the probability that certain risk factors would occur in certain groups of African American and white women in Mississippi. METHODS: We extracted self-reported data on 9690 women in Mississippi from the BRFSS for 1991 to 1999. We performed logistic regression analyses to examine the independent effects of age and race as relative risks for cardiovascular disease, controlling for socioeconomic status. RESULTS: Odds ratios for cigarette smoking for some African American women, for high blood pressure for some white women, and for diabetes in some white and African American women were significantly elevated. Prevalence data, however, showed that African American women had a higher prevalence of diabetes and overweight than white women did. CONCLUSION: The prevalence of overweight among Mississippi women seemed to increase as they aged. Decreasing obesity should be a major public health priority, as obesity's association with several chronic diseases is well documented and it is very prevalent among Mississippi women. Implementing programs to address unhealthy behaviors is essential for maintaining good health and thus should be strongly encouraged. PMID- 12744426 TI - Patient choice of provider gender. AB - As the proportion of women physicians in the United States increases, patients have increased access to physicians of either sex, and some patients express a clear preference for female providers. This is especially true in obstetrics/gynecology, where patients may have a variety of reasons for requesting female physicians. This column presents a case in which the patient not only expressed a preference for a female physician, but also, in fact, refused care from any male obstetrician/gynecologist. Possible responses to such a request are examined, with consideration of the competing priorities involved. PMID- 12744425 TI - Women's knowledge and sources of information on the risks and benefits of oral contraception. AB - OBJECTIVE: To assess the knowledge of the risks and benefits of oral contraceptives (OCs) in a heterogeneous group of women and to identify their sources of information. METHODS: A self-administered questionnaire assessing demographics, contraception history, knowledge of risks and benefits of OCs, and information sources was given to literate English- and Spanish-speaking women waiting for appointments at 4 clinics serving distinct populations in Portland, Oregon. RESULTS: Approximately half of the 211 women studied were of the opinion that OCs decreased the risk of acne, dysmenorrhea, and menorrhagia and increased the risk of weight gain, headaches, and thrombosis. Less than 15% knew of the decreased risk of anemia, endometrial cancer, colon cancer and pelvic inflammatory disease, but 28% understood the decreased risk of ovarian cancer. Seven percent to 36% of women used their own experiences in assessing the effect of OCs on a variety of general and reproductive factors. Women relied primarily on printed information for knowledge of OCs' effects on cardiovascular health and cancer. CONCLUSION: Women in this heterogeneous population of women were unaware of several benefits of OCs. Women relied heavily on their own experiences in assessing the risks and benefits of OCs. Women cited printed information more frequently than medical personnel as major sources of information on cardiovascular and oncological risks and benefits of OCs. The Internet, however, played a minimal, if any role in educating women about OCs. PMID- 12744427 TI - Rx for negotiating. PMID- 12744428 TI - Geostatistical ground water monitoring of a point source NO3(-)-N plume entering a restored riparian zone. AB - NO3(-)-N concentrations associated with a point-source ground water plume were monitored as the plume entered a restored riparian zone (RRZ) on the University of Idaho campus in Moscow, Idaho. Seasonal NO3(-)-N data collected for a network of piezometers installed in the RRZ were analyzed geostatistically and then modeled spatially using the conditional simulation method of sequential Gaussian simulation (SGS). SGS was utilized to predict the spatial distribution of ground water NO3(-)-N concentrations at unsampled locations in the RRZ and to quantify the uncertainty associated with the prediction. Maps prepared using SGS results illustrate the short-scale variability, or patchiness, expected of NO3(-)-N concentration distributions in a riparian zone. Manipulation of SGS output provided graphical and quantitative estimates of the likelihood of exceeding a specified NO3(-)-N concentration threshold at a given confidence level for any location within the RRZ. Geostatistical simulation tools for quantifying uncertainty also provide a potential risk assessment methodology for making remediation decisions and reducing remediation costs. PMID- 12744429 TI - Charged body potential monitoring of an electrolyte plume emanating from a dripping source. AB - Hole-surface charged body potential (CBP) measurements were taken over a 173-day period during a drip-injection, tracer experiment in partially saturated, fractured basalt. A continuous, enhanced conductivity, potassium chloride (KCl) solution was dripped into the fractured basalt and energized directly through a current electrode placed in the conductive solution. The constant concentration, KCl solution was introduced above a perched water table at an average rate of 10.07 L/day under a constant hydraulic head for 76 days. The KCl drip period was followed by a 34-day tap water drip period and a 62-day drainage period. Hole surface CBP measurements were taken over time to delineate the evolution of the asymmetrical, vadose zone, plume. A 15 by 15 grid of land surface based, porous pot electrodes (copper sulfate), located symmetrically about the centrally located injection borehole, was used for the hole-surface CBP experiment. Ratios of electrical potentials measured at the land surface over time were contoured and profiled to delineate the evolution of the electrolyte plume. PMID- 12744430 TI - A review of riverine fluxes of hexachlorocyclohexane and DDT to the Azov and Black seas from the former USSR and Russian Federation. AB - While the Azov and Black seas are subject to anthropogenic pollution to a much higher extent than any other seas, this has been little studied with only a few critical reviews of contaminant fluxes to these seas. Riverine fluxes of the organochlorine pesticides (OCPs) hexachlorocyclohexane (HCH) and DDT to the Azov and Black seas from the former Soviet Union and Russian Federation were thus reviewed for 1986 to 1996. The review was based on official data and data obtained by independent specialists. The amount of HCH used, and the intensity of usage, in these river catchments decreased during the review period. Concurrently, OCP concentrations in the rivers and their fluxes also decreased according to both official and independent data. A comparison of the official and the independent data sets for 1988 revealed significant differences, reflecting the need for more rigorous sampling and analytical protocols for both data sets. According to the OGSNK/GSN data, the flux rates of the five largest rivers were ranked (from largest to smallest) as follows: Don > Dnestr > Danube > Kuban > Dnepro (alpha-HCH); Danube > Don > Dnestr > Dnepro > Kuban (gamma-HCH); Dnestr > Danube > Don > Dnepro-Kuban (DDT+DDE). For rivers with lower annual riverine discharges, the DDT fluxes were surprisingly high (0.43 to 1.49 tonnes a(-1)). According to independent data for 1988 the rankings of the rivers was: Danube > Don > Dnepro > Dnestr > Kuban (alpha-HCH); Danube > Don > Dnestr > Dnepro > Kuban (gamma-HCH); Danube > Dnepro > Dnestr > Don > Kuban (DDT); Danube > Dnepro > Don > Kuban > Dnestr (DDE). The DDT flux estimates for small rivers derived from independent data were 19 to 46 times lower than those calculated using OGSNK/GSN data. According to the independent data, the total riverine OCP transport from the Russian Federation into the Azov Sea from 1988 to 1996 was 1.288 tonnes of gamma-HCH+alpha-HCH and 1.693 tonnes of DDT+DDE while for the Black Sea they were 3.830 tonnes and 5.116 tonnes for gamma-HCH+alpha-HCH and DDT+DDE, respectively. PMID- 12744431 TI - Complexation of butylbenzenes with 2-hydroxypropyl-cyclodextrins in aqueous solution. AB - The stability constants for the inclusion of butylbenzenes (BBs) with 2 hydroxypropyl-cyclodextrins (2HP-CDs) in aqueous solution have been determined using the static head-space method. The 1:1 stability constants were obtained by this method. For the complexation of BBs by 2HP-alpha-CD, the order of stability constants was n-BB > isoBB > tert-BB. In contrast, the order for complexation of BBs by 2HP-beta-CD was the opposite: tert-BB > isoBB > n-BB. The order of the stability constants by 2HP-gamma-CD was similar to that of 2HP-beta-CD, but the values of the stability constants were not significantly different in the BB types. In addition, the binding effects are discussed in relation to the different alkyl groups of BBs. PMID- 12744433 TI - Changes in the content of polycyclic aromatic hydrocarbons (PAHs) in light soil fertilized with sewage sludge. AB - In the present work the influence of soil fertilization with sewage sludge on the changes in the content of polycyclic aromatic hydrocarbons (PAH) in this soil was studied during a one-year field experiment. The experiment consisted of six plots to which sewage sludge was introduced at doses of 30 (1%), 75 (2.5%), 150 (5%), 300 (10%) and 600 ton/ha (20%) as calculated for dry mass. The PAH content at a depth of 0-20 and 20-40 cm was determined. With time, an increase in the PAH content in the lower soil layer (20-40 cm) and a gradual decrease of the PAH content in the 0-20 cm layer was observed. In most cases, a decrease by half in the PAH content in the soil fertilised with sewage sludge was observed. PMID- 12744432 TI - Determination of polycyclic aromatic hydrocarbons in indoor and outdoor air with chromatographic methods. AB - Polycyclic aromatic hydrocarbons in indoor and outdoor air of four typical homes in Hangzhou, China were determined with a highly automated chromatographic method. The results indicated that the concentrations of the 15 polycyclic aromatic hydrocarbons in the indoor air were between 1.907 microg/m3 and 14.29 microg/m3, which were much higher than those in the corresponding outdoor air. Because of the popular use of mothball in wardrobes, naphthalene had the highest concentration in all the 15 polycyclic aromatic hydrocarbons, which was up to 13.17 microg/m3 and contributed 68% to the total polycyclic aromatic hydrocarbons in the room. In kitchen, because of the representative cooking method, there had much of three- and four-ring polycyclic aromatic hydrocarbons. The indoor smoking not only led to high concentration of polycyclic aromatic hydrocarbons in the indoor air, but also contributed almost all the benzo(a)pyrene. Naphthalene, acenaphthene and pyrene were considered being generated by the indoor sources in all the four homes. PMID- 12744434 TI - Elemental transport and distribution in soils amended with incinerated sewage sludge. AB - Sewage sludge (SS) is the major solid waste of sewage and wastewater treatment plants in cities around the world. Even though treated effluent water from wastewater treatment plants are utilized for irrigation, disposal of sewage sludge is becoming a serious problem. This is due to its high content of certain heavy metals still posing threat of accumulation in plants and groundwater contamination when it is used as soil amendment or disposed in landfills. Water treatment plants incinerate the dewatered activated sewage sludge (ISS) and dissolve the ash in water to store in ash ponds for long-term storage (WISS). A study was undertaken to evaluate the transport and leaching potential of various elements and their distribution within soil columns amended with various rates of ISS. Results of this study indicates that ISS from wastewater treatment plants can be used as soil amendment on agricultural lands at low to medium rates (< or = 100 Mg ha(-1)) without causing potential loading of metals into groundwater. PMID- 12744436 TI - Heavy metal pollution in the soils of various land use types based on physicochemical characteristics. AB - In this study, soil samples were collected at eight different regional types of Seoul City and analyzed for their physicochemical properties. In addition, the distribution of heavy metal concentrations was analyzed using samples representing both the surface and deep soil layer. The physicochemical properties analyzed for those samples included parameters such as pH, moisture content, apparent (and true) density, pore ratio, solid content, conductivity, ionic strength, total dissolved solid (TDS), total organic carbon (TOC), and total phosphorus (TP). The contents of heavy metal components contained in plant leaves were also analyzed and compared with those measured from different soil layers. Contents of Cu and Cd were highest in the DH area among eight locations investigated and Pb was higher in the surface soil samples of the GS region than any other locations. According to physicochemical properties of the surface and deep soils, acidity was higher in the surface than deep soils. Depending on the selection of treatment method between strong and weak acids, the metal concentrations were larger by 3-5 times in the strong acid than the weak acid treatments. In addition, metals were higher in the deep than in the surface soil and relative metal contents of leaf samples closely resembled those of soil samples. Results of this study suggest that the physicochemical properties of soils determined from different regional types of Seoul area exhibited a close relationship with the land use types and environmental conditions surrounding each region. PMID- 12744435 TI - Soil contamination and plant uptake of heavy metals at polluted sites in China. AB - We investigated heavy metal contamination in soils and plants at polluted sites in China including some with heavy industries, metal mining, smelting and untreated wastewater irrigation areas. We report our main findings in this paper. The concentrations of heavy metals, including Cd and Zn, in the soils at the investigated sites were above the background levels, and generally exceeded the Government guidelines for metals in soil. The concentrations of metals in plants served to indicate the metal contamination status of the site, and also revealed the abilities of various plant species to take up and accumulate the metals from the soil. Substantial differences in the accumulation of heavy metals were observed among the plant species investigated. Polygonum hydropiper growing on contaminated soils in a sewage pond had accumulated 1061 mg kg(-1) of Zn in its shoots. Rumex acetosa L. growing near a smelter had accumulated more than 900 mg kg(-1) of Zn both in its shoots and roots. Therefore these species have potential for phytoremediation of metal-contaminated sites. Our results indicate the need to elucidate the dynamics of soil metal contamination of plants and the onward movement of metal contaminants into the food chain. Also our results indicate that the consumption of rice grown in paddy soils contaminated with Cd, Cr or Zn may pose a serious risk to human health, because from 24 to 22% of the total metal content in the rice biomass was concentrated in the rice grain. Platanus acerifolia growing on heavily contaminated soil accumulated only very low levels of heavy metals, and this mechanism for excluding metal uptake may have value in crop improvement. Sources of metal entering the environmental matrices studied included untreated wastewater, tailings or slurries and dust depositions from metal ore mining, and sewage sludge. Pb, Zn or Cd concentrations declined with the distance from metal smelter in accordance with a good exponential correlation (R2>0.9), and this shows that metal dust deposition is an important contributor to metal contamination of soils. PMID- 12744437 TI - Benthic algae as monitors of heavy metals in various polluted rivers by Energy Dispersive X-Ray Spectrometer. AB - Benthic microalgae assemblages were used as monitors of copper (Cu), zinc (Zn), and chromium (Cr) in various polluted rivers of San-Yeh-Kong, in southern Taiwan, and analyzed using Scanning Electron Microscope and Energy Dispersive X-Ray Spectrometer (SEM-EDS). Under SEM-EDS, the benthic algae from seriously polluted rivers (dominant by the cyanobacteria Oscillatoria chalybea, green algae Euglena acus and diatom Nitzschia palea under light microscopes) revealed the elemental compositions of heavy metals such as Cu, Zn, Cr, Ti, and that of Mg, Al, Si, P, S, Cl, K, Ca, and Fe. In contrast, benthic algae from moderately (dominant by diatoms Cymbella turgidula and Gomphonema globiferm) and lightly polluted river (dominant by diatom Diatoma vulgare) didn't have any heavy metal elements. In addition to the algal samples, unfiltered water and bottom mud were also investigated for comparison. Further experiment involving the benthic algae from seriously polluted station revealed that all dominant species could survive on the 1 mL 60 ppm Cu, 1 mL 60 ppm Zn, and 1 mL 60 ppm Cr separately after five days culture. The data of this preliminary study are sufficient to encourage further experimentation into the potential for detecting benthic algae as a bioindicator under SEM-EDS to provide rapid information about water pollution. PMID- 12744438 TI - Spatial and temporal variabilities of nutrient limitation based on in situ experiments of nutrient enrichment bioassay. AB - In situ experiments of Nutrient Enrichment Bioassays (NEBs) were performed in a morphologically complex reservoir of a temperate region to identify primary limiting nutrient regulating phytoplankton productivity and determine a severeity of the nutrient limitation. Absolute nutrient contents and nutrient ratios of TN:TP and TDN:TDP in the ambient water indicated a potential limitation of phosphorus. This outcome agreed with the results of NEBs; Algal response in the P treatments, expressed as a ratio of CHLf:CHLi, showed significantly (p < 0.05) greater algal response than P + NH4-N or P + NO3-N treatments. The magnitude of the limitation, however, showed large spatial and temporal variations. The response in treatments enriched with phosphorus (P, P + NH4-N, and P + NO3-N) was greatest in the downlake zone and least in the point-source zone, while the response was greater during summer monsoon than any other seasons. Algal growth rate experiments showed that the response in treatments enriched with NO3-N and P + NO3-N never exceeded > 0.50 microg L(-1) per day and was significantly (p < 0.05) less than that in the three treatments with P (1P, 2P, and P + NH4-N), indicating a reduced response in the treatments enriched with NO3-N. The suppression of nitrate on algal growth seemed to be involved with inhibition effect of nitrogenase activity through low-nitrate uptake rate in the high nitrate-rich environment and large additions of potassium with phosphorus in spike. Regression analysis of log10-transformed CHLf:CHLi ratios against ambient nutrient contents showed that in situ algal response in the P treatments was inversely related (r = -0.77; p < 0.001) with ambient TP and positively correlated (r = 0.75; p < 0.001) with TN:TP ratios. This result suggests that in situ response depends largely on external loading of limiting nutrients. PMID- 12744439 TI - Factors affecting the performances of sepiolite and zeolite for the treatment of textile wastewater. AB - In this study, the adsorption mechanism of three reactive azo dyes (Everzol Black B, Everzol Red 3BS, Everzol Yellow 3RS H/C) on natural porous minerals, sepiolite and zeolite, have been examined in order to identify the ability of clay minerals on the removal of colored textile dyes from aqueous solutions. High ion exchange capacity, high surface areas and importantly their relatively cheap prices make them attractive adsorbents. For this purpose, series of batch adsorption tests were carried out as a function of solids concentration, time and dye concentration using natural and modified sepiolites and zeolites. Also, experiments were conducted to determine the amount of solid to be used in adsorption tests. An optimum solids concentration was selected as 0.05 g/mL (5%). Most of the adsorption was found to take place within the first 2h of mixing, taking into account the extreme changes like pH and concentration, the mixing period was selected as 4 h for further testing. The adsorption results indicate that both natural sepiolite and zeolite have limited adsorption capacities of the reactive dyes but are substantially improved by modifying their surfaces with quaternary amines. The mechanism of adsorption process is elucidated on the basis of experimental data. PMID- 12744440 TI - Electrophoretic nature and evaluation of poly-aluminum-chloride-sulfate (PACS) as a coagulant for water and wastewater treatment. AB - A series of poly-aluminum-chloride-sulfates (PACS), which have different OH/Al (gamma) and Al(3+)/SO4(2-) mole ratios, has been prepared using AlCl3 x 6H2O, A(SO4)3 x 18H2O and Na2CO3 as raw materials. The electrophoretic nature of PACS was investigated by electrophoresis. Laboratory experiments were undertaken to evaluate the PACS in comparison with polyaluminum chloride (PAC) for the coagulation of simulating water and actual wastewaters. The experimental results show that the gamma value and the Al(3+)/SO4(2-) mole ratio affect the electrophoretic nature of PACS. PACS has a maximum zeta potential at about a gamma value of 1.5 and Al(3+)/SO4(2-) mole ratio of 12-16. The zeta potential of PACS varies with pH. The performance of PACS as coagulant is affected by gamma value and Al(3+)/SO4(2-) mole ratio. PACS of gamma = 2.1 and Al(3+)/SO4(2-) = 15 gives best turbidity removal effectiveness. In comparison with PAC of gamma = 2.0, PACS of gamma = 2.0 and Al(3+)/SO4(2-) = 16 gives higher removal efficiency for turbidity and COD, and shows the following advantages in the clarification of waters and wastewaters: rapid aggregation velocity, larger and heavier flocs, and lower required dosage. PMID- 12744441 TI - Leaching behavior and ESEM characterization of water-sensitive mudstone in southwestern Taiwan. AB - This investigation attempts to understand the critical soluble salts in natural mudstone and the leaching, microstructural, and microchemical characteristics in soaked mudstone using scanning electron microscopy (SEM)/energy-dispersive X-ray analysis (EDAX), X-ray fluorescence spectrometry (XRF), X-ray diffractometry (XRD), conductivity measurement, ion chromatography (IC), and environmental scanning electron microscopy (ESEM)/EDAX techniques. Natural mudstone probably includes soluble salts such as Na2SO4, NaCl, NaCO3, and CaCO3. The dissolution of Na2SO4 controls water-sensitive mudstone very susceptible to slaking and dispersion. ESEM micrographs clearly show evidence of mudstone-slaking during soaking since the visible pores are filled with small aggregative masses. A calcium-bearing precipitate from the soaked mudstone is speculated to be attributable to the decomposition of the hydrated product of the fresh mudstone. PMID- 12744442 TI - Counterflow co-flocculation flotation for water purification. AB - A new method for potable water treatment was brought forward and studied in this research. The treatment process was named as counterflow co-flocculation flotation (CC-FF). Pilot experiment was conducted and the operational parameters were presented. The optimized operational conditions are as follows: the detention time is 6-11 min with hydraulic load of 9-16 m3/(m2 h); the recycle ratio should be no less than 8% while the distance between the inlet of source water and recycle water should be greater than 1200 mm. If the source water turbidity was lower than 100 NTU, 0.12-0.35 mmol/L Al dosage is enough to maintain efficient turbidity removal. Since the flocculation and flotation processes were carried out in the same tank, this new technique has some advantages than the conventional flocculation-flotation methods. Firstly, the microbubbles released from recycle water will participate in the flocculation of suspended particles, hence the low-density but high shear-force-resistance flocci could be formed. Secondly, the microflocci or suspended particles will be functioned as 'nucleus' during the bubble formation from air-dissolved recycle water. Thirdly, in the midsection of the tank a blanket of bubble-microfloc aggregates could be formed, which will intercept the downward-flow flocci and upward-flow bubbles efficiently, thus keep the renovation and stability of the blanket. PMID- 12744443 TI - Performance of COD removal from acid scarlet BS-containing solution in a novel packed-bed hollow-tube photocatalytic (PHP) reactor. AB - A novel packed-bed hollow-tube photocatalytic (PHP) reactor using TiO2-coated Ti particles as fillers was designed and applied to treat a simulated dye wastewater containing Acid Scarlet BS. The experimental results showed that PHP reactor could efficiently remove chemical oxygen demand (COD) from the dye solution and the COD removal efficiency was considerably dependent on the operating parameters, airflow, initial dye concentration and initial pH value of solution. It was also found that the inserting of the hollow tubes could apparently increase the COD removal efficiency of the packed-bed photo-reactor while the application of external electric field could improve the degradation efficiency of the dye but not obviously promote COD removal. PMID- 12744444 TI - Towards understanding the TiO2-mediated photoredox process of Cu(II)-formic acid solution. AB - The performance of TiO2-film was investigated in the process of simultaneous photocatalytic Cu(II) reduction and COD (chemical oxygen demand) removal from Cu(II)-containing formic acid solution during successive batch runs. It was found that, during the first batch run, the Cu(II) ions could increase the COD removal efficiency, synchronous to the Cu(0) deposition, however, decrease the efficiency by a factor of 22.5% with the continuative proceeding of the deposition for the second batch run. Unexpectedly, after the second run the COD removal efficiency remained a constant, ca. 40%, and no longer continued decrease. The unexpected observation was attributed to the cycle of Cu(II) deposition and Cu(0) re dissolution due to air oxidation at pH 2.73. The attribution was primarily confirmed by the scanning electron microscope image and several fluctuation phenomena in the reaction process, such as pH, Cu(II) concentration and photocurrent. PMID- 12744445 TI - Accumulation and elimination of cadmium and zinc in the Asian periwinkle Littorina brevicula. AB - To elucidate the differences between the detoxification mechanisms of essential metal (Zn) and non-essential metal (Cd) in Littorina brevicula that is highly resistant to a wide range of heavy metal concentrations, Asian periwinkles were exposed to Cd (400 microg/L), Zn (3000 microg/L) and a mixture of both metals. We examined metal accumulation, elimination and subcellular distribution for binding to proteins. The metal concentration in L. brevicula increased gradually with exposure time (up to 70 days), following which accumulated levels reached saturation point. The accumulated Zn content was increased in the presence of Cd, while Cd uptake was decreased when Zn was present. During the depuration period (42 days), Cd was not removed from periwinkles, while Zn was eliminated in a rate of 2.19 microg Zn g(-1) day(-1). This elimination rate was particularly high on exposure to the metal mixture (3.80 microg Zn g(-1) day(-1)). Subcellular distribution studies on Cd and Zn revealed that most Cd (80%) was bound to cytosolic ligand, while more than 75% Zn was distributed in the membrane (insoluble) fraction. An additional difference in sequestering of metal in the cytosol was noted between Cd and Zn; most Cd in the cytosol was bound to metallothionein-like cadmium binding protein, MBP-1 (9.8 kDa), while the profile for Zn distribution revealed the presence of four Zn-binding ligand peaks, specifically, HMW (60 kDa), MBP-1 (9.8 kDa), MBP-2 (5 kDa) and LMW (<1 kDa). Our data confirm that metallothionein-like cadmium binding protein, MBP-1, has the same affinity for Zn, while MBP-2 displays comparatively higher affinity for Zn than Cd. PMID- 12744446 TI - Experimental testing of spray dryer for control of incineration emissions. AB - The research investigated the absorption/adsorption efficiency of sulfur dioxide (SO2), heavy metals, and polycyclic aromatic hydrocarbons (PAHs) with different Ca-based sorbents in a spray dryer during incineration process. For further improving the adsorption capacity of Ca-based sorbents, different spraying pressure and additives were carried out in this study. Experimental results showed that CaO could be used as an alternative sorbent in the spray dryer at an optimal initial particle size distribution of spraying droplet. In the spray dryer, Ca-based sorbents provided a lot of sites for heavy metals and PAHs condensing and calcium and alkalinity to react with metals to form merged species. As a result, heavy metals and PAHs could be removed from the flue gas simultaneously by condensation and adsorption. The additions of additives NaHCO3, SiO2, and KMnO4 were also found to be effective in improving the removal efficiency of these air pollutants. PMID- 12744447 TI - Media attitudes vs. use: the contribution of context to the communication environment in telemedicine. AB - Media attitudes and media use have been the focus of considerable academic research. This article uses this research to explore patient and health care practitioner attitudes toward telemedicine interactions via videoconferencing technology. PMID- 12744448 TI - Rates of return from hospital conversions. AB - This article presents information on the rates of return obtained by purchasers of U.S. hospitals since the mid 1980s. The key hypothesis tested in this study was whether for-profit acquirers are able to purchase hospitals at below-market prices. We test the hypothesis by comparing internal rates of return to an estimate of the weighted cost of capital for all for-profit hospitals in the year the transaction occurred. PMID- 12744449 TI - Outcomes of physician job satisfaction: a narrative review, implications, and directions for future research. AB - The decline in physician job satisfaction is well documented, but its impact on physicians and patients is not. This article reviews 44 articles that address these impacts. Four findings emerge from this study along with recommendations for improving the measurement and management of job satisfaction. PMID- 12744450 TI - Effect of hospital conversion on organizational decision making and service coordination. AB - This study looks into the "black box" of hospital ownership conversion in a "natural experiment." We posed two competing theories about how conversion might influence management practices. Results support complexity theory and not threat rigidity theory. As predicted from complexity theory, MDs and RNs had greater levels of participation and influence over final decision choices in converted hospitals than in nonconverted hospitals. PMID- 12744451 TI - Introduction to The Forum.: health care delivery system in rural communities. PMID- 12744452 TI - The evolution of rural health networks: implications for health care managers. AB - This article examines the development and operation of rural health networks in the United States based on data collected from telephone surveys of rural health networks containing at least one rural hospital in the United States in 1996 and four years later in 2000. The implications of network development for health care managers participating in, or considering participation in, a rural health network are discussed. PMID- 12744453 TI - Strategy as simple rules: understanding success in a rural clinic. AB - Drawing on chaos and complexity theories, we apply the concept of "strategy as simple rules" to analyze the success of one rural health clinic. We document how such rules may emerge as success is viewed retrospectively. We describe four simple rules that emerged in this setting and explicate how our case analysis may assist other rural providers. PMID- 12744454 TI - Developing rural primary health care service areas: an analysis of patient migration patterns. AB - This study explores the potential of health care service areas (HCSAs) developed from claims data as the geopolitical units for the assessment of health professional needs in rural areas. HCSAs reflect consumer shopping patterns for health care and alter traditional health professional needs estimates. PMID- 12744455 TI - The sypA, sypS, and sypC synthetase genes encode twenty-two modules involved in the nonribosomal peptide synthesis of syringopeptin by Pseudomonas syringae pv. syringae B301D. AB - Syringopeptin is a necrosis-inducing phytotoxin, composed of 22 amino acids attached to a 3-hydroxy fatty acid tail. Syringopeptin, produced by Pseudomonas syringae pv. syringae, functions as a virulence determinant in the plant-pathogen interaction. A 73,800-bp DNA region was sequenced, and analysis identified three large open reading frames, sypA, sypB, and sypC, that are 16.1, 16.3, and 40.6 kb in size. Sequence analysis of the putative SypA, SypB, and SypC sequences determined that they are homologous to peptide synthetases, containing five, five, and twelve amino acid activation modules, respectively. Each module exhibited characteristic domains for condensation, aminoacyl adenylation, and thiolation. Within the aminoacyl adenylation domain is a region responsible for substrate specificity. Phylogenetic analysis of the substrate-binding pockets resulted in clustering of the 22 syringopeptin modules into nine groups. This clustering reflects the substrate amino acids predicted to be recognized by each of the respective modules based on placement of the syringopeptin NRPS (nonribosomal peptide synthetase) system in the linear (type A) group. Finally, SypC contains two C-terminal thioesterase domains predicted to catalyze the release of syringopeptin from the synthetase and peptide cyclization to form the lactone ring. The syringopeptin synthetases, which carry 22 NRPS modules, represent the largest linear NRPS system described for a prokaryote. PMID- 12744456 TI - Characterization and expression analysis of genes encoding phosphoenolpyruvate carboxylase and phosphoenolpyruvate carboxylase kinase of Lotus japonicus, a model legume. AB - Phosphoenolpyruvate carboxylases (PEPCs), one form of which in each legume species plays a central role in the carbon metabolism in symbiotic root nodules, are activated through phosphorylation of a conserved residue by a specific protein kinase (PEPC-PK). We characterized the cDNAs for two PEPC isoforms of Lotus japonicus, an amide-translocating legume that forms determinate nodules. One gene encodes a nodule-enhanced form, which is more closely related to the PEPCs in amide-type indeterminate nodules than those in ureide-type determinate nodules. The other gene is expressed in shoots and roots at a low level. Both forms have the putative phosphorylation site, Ser11. We also isolated a cDNA and the corresponding genomic DNA for PEPC-PK of L. japonicus. The recombinant PEPC PK protein expressed in Escherichia coli phosphorylated recombinant maize C4-form PEPC efficiently in vitro. The level of mRNA for PEPC-PK was high in root nodules, and those in shoots and roots were also significant. In situ hybridization revealed that the expression patterns of the transcripts for PEPC and PEPC-PK were similar in mature root nodules, but were different in emerging nodules. When L. japonicus seedlings were subjected to prolonged darkness and subsequent illumination, the activity of PEPC-PK and the mRNA levels of both PEPC and PEPC-PK in nodules decreased and then recovered, suggesting that they are regulated according to the amounts of photosynthates transported from shoots. PMID- 12744457 TI - The Arabidopsis genes RPW8.1 and RPW8.2 confer induced resistance to powdery mildew diseases in tobacco. AB - Plant disease resistance (R) gene products recognize pathogen avirulence (Avr) gene products and induce defense responses. It is not known if an R gene can function in different plant families, however. The Arabidopsis thaliana R genes RPW8.1 and RPW8.2 confer resistance to the powdery mildew pathogens Erysiphe orontii, E. cichoracearum, and Oidium lycopersici, which also infect plants from other families. We produced transgenic Nicotiana tabacum, N. benthamiana, and Lycopersicon esculentum plants containing RPW8.1 and RPW8.2. Transgenic N. tabacum plants had increased resistance to E. orontii and O. lycopersici, transgenic N. benthamiana plants had increased resistance to E. cichoracearum, but transgenic L. esculentum plants remained susceptible to these pathogens. The defense responses induced in transgenic N. tabacum and N. benthamiana were similar to those mediated by RPW8.1 and RPW8.2 in Arabidopsis. Apparently, RPW8.1 and RPW8.2 could be used to control powdery mildew diseases of plants from other families. PMID- 12744458 TI - Members of the Arabidopsis WRKY group III transcription factors are part of different plant defense signaling pathways. AB - WRKY proteins are a large group of transcription factors restricted to the plant kingdom. In Arabidopsis thaliana, the gene family consists of 74 members. Here, we analyzed the expression of all 13 members of one main WRKY subgroup and found that the majority are responsive both to pathogen infection and to salicylic acid. Temporal expression studies during compatible, incompatible, and nonhost interactions and employing plant defense-signaling mutants allowed us to define four distinct WRKY subsets responding to different signaling queues along defense pathways. These subsets did not reflect phylogenetic relationships. Promoter studies of one member, AtWRKY54, using a reporter gene construct in transgenic Arabidopsis plants, revealed that regulatory regions mediating pathogen and SA inducibility are clearly separable. In an AtWRKY54 knockout line, resistance to Peronospora parasitica was not compromised, but the transient expression kinetics of several WRKY genes was affected, suggesting both the existence of functional redundancy and intense cross-talk between signaling networks. PMID- 12744460 TI - The Colletotrichum lagenariu Ste12-like gene CST1 is essential for appressorium penetration. AB - Colletotrichum lagenarium is the causal agent of anthracnose of cucumber. This fungus produces a darkly melanized infection structure, appressoria, to penetrate the host leaves. The C. lagenarium CMK1 gene, a homologue of the Saccharomyces cerevisiae FUS3/KSS1 mitogen-activated protein (MAP) kinase genes, was shown to regulate conidial germination, appressorium formation, and invasive growth. In S. cerevisiae, Ste12p is known to be a transcriptional factor downstream of Fus3p/Kss1p MAP kinases. To evaluate the CMK1 MAP kinase pathway, we isolated the Ste12 homologue CST1 gene from C. lagenarium and characterized. The cst1delta strains were nonpathogenic on intact host leaves, but could form lesions when inoculated on wounded leaves. Conidia of the cst1delta strains could germinate and form melanized appressoria on both host leaf surface and artificial cellulose membrane, but could not produce infectious hyphae from appressoria, suggesting that CST1 is essential for appressorium penetration in C. lagenarium. In addition, matured appressoria of the cst1delta strains contained an extremely low level of lipid droplets compared with that of the wild-type strain. Lipid droplets were abundant in conidia of the cst1delta strains, but rapidly disappeared during appressorium formation. This misscheduled lipid degradation might be related to the failure of appressorium penetration in the cst1delta strain. PMID- 12744461 TI - The role of nod factor substituents in actin cytoskeleton rearrangements in Phaseolus vulgaris. AB - In order to define the symbiotic role of some of the chemical substituents in the Rhizobium etli Nod factors (NFs), we purified Nod metabolites secreted by the SM25 strain, which carries most of the nodulation genes, and SM17 with an insertion in nodS. These NFs were analyzed for their capabilities to induce root hair curling and cytoskeletal rearrangements. The NFs secreted by strain SM17 lack the carbamoyl and methyl substituents on the nonreducing terminal residue and an acetyl moiety on the fucosyl residue on the reducing-terminal residue as determined by mass spectrometry. We have reported previously that the root hair cell actin cytoskeleton from bean responds with a rapid fragmentation of the actin bundles within 5 min of NF exposure, and also is accompanied by increases in the apical influxes and intracellular calcium levels. In this article, we report that methyl-bearing NFs are more active in inducing root hair curling and actin cytoskeleton rearrangements than nonmethylated NFs. However, the carbamoyl residue on the nonreducing terminal residue and the acetyl group at the fucosyl residue on the reducing terminal residue do not seem to have any effect on root hair curling induction or in actin cytoskeleton rearrangement. PMID- 12744462 TI - Rhizobium-lnduced calcium spiking in Lotus japonicus. AB - Legumes and rhizobium bacteria form a symbiosis that results in the development of nitrogen-fixing nodules on the root of the host plant. The earliest plant developmental changes are triggered by bacterially produced nodulation (Nod) factors. Within minutes of exposure to Nod factors, sharp oscillations in cytoplasmic calcium levels (calcium spiking) occur in epidermal cells of several closely related legumes. We found that Lotus japonicus, a legume that follows an alternate developmental pathway, responds to both its bacterial partner and to the purified bacterial signal with calcium spiking. Thus, calcium spiking is not restricted to a particular pathway of nodule development and may be a general component of the response of host legumes to their bacterial partner. Using Nod factor-induced calcium spiking as a tool to identify mutants blocked early in the response to Nod factor, we show that the L. japonicus Ljsym22-1 mutant but not the Ljsym30 mutant fails to respond to Nod factor with calcium spiking. PMID- 12744463 TI - New NodW- or NifA-regulated Bradyrhizobium japonicum genes. AB - A cluster of genes coding for putative plant cell-wall degrading enzymes (i.e., genes for two endoglucanases [gunA and gunA2], one pectinmethylesterase [pme], and one polygalacturonase [pgl]) was identified by sequence similarities in the symbiotic region of the Bradyrhizobium japonicum chromosome. In addition, a systematic screen of the region revealed several genes potentially transcribed by the sigma(54)-RNA polymerase and activated by the transcriptional regulator NifA (i.e., genes for proteins with similarity to outer membrane proteins [id117 and id525] and a citrate carrier [id331 or citA] and one open reading frame without similarity to known proteins [id747]). Expression studies using transcriptional lacZ fusions showed that gunA2 and pgl were strongly induced by the isoflavone genistein in a NodW-dependent manner, suggesting a role of the gene products in early events of the nodulation process; by contrast, gunA and pme expression was very weak in the conditions tested. The gunA2 gene product was purified and was shown to have cellulase activity. beta-Galactosidase activity expressed from transcriptional lacZ fusions to id117, id525, and id747 in the wild type and in nifA and rpoN mutant backgrounds confirmed that their transcription was dependent on NifA and sigma(54). Despite the presence of a -24/-12-type promoter and a NifA binding site upstream of citA, no regulation could be demonstrated in this case. Null mutations introduced in gunA, gunA2, pgl, pme, citA, id117, id525, and id747 did not impair the symbiosis with the host plants. PMID- 12744464 TI - Characterization of a novel barley protein, HCP1, that interacts with the Brome mosaic virus coat protein. AB - Brome mosaic virus (BMV) requires the coat protein (CP) not only for encapsidation but also for viral cell-to-cell and long-distance movement in barley plants. This suggests that BMV infection is controlled by interactions of CP with putative host factors as well as with viral components. To identify the host factors that interact with BMV CP, we screened a barley cDNA library containing 2.4 x 10(6) independent clones, using a yeast two-hybrid system. Using full-length and truncated BMV CPs as baits, four candidate cDNA clones were isolated. One of the candidate cDNAs encodes a unique oxidoreductase enzyme, designated HCP1. HCP1 was found predominantly in the soluble fractions after differential centrifugation of BMV-infected and mock-inoculated barley tissues. A two-hybrid binding assay using a series of truncated BMV CPs demonstrated that a C-terminal portion of CP is essential for its interaction with HCP1. Interestingly, experiments with CP mutants bearing single amino acid substitutions at the C-terminus revealed that the capacity for mutant CP-HCP1 binding correlates well with the infectivity of the corresponding mutant viruses in barley. These results indicate that CP-HCP1 binding controls BMV infection of barley, interacting directly with CP, probably in the cell cytoplasm. PMID- 12744465 TI - Disruption of Botrytis cinerea pectin methylesterase gene Bcpme1 reduces virulence on several host plants. AB - The pectinolytic enzyme pectin methylesterase (PME) hydrolyses pectin in methanol and polygalacturonic acid. In the expressed sequence tag library of Botrytis cinerea T4, we identified a 1,041 bp Bcpme1 cDNA potentially encoding a 346-amino acid protein of 37 kDa showing 46.8% identity with Aspergillus sp. PMEs. Bcpme1 is a single copy gene and is similarly expressed in glucose and pectin containing media. To evaluate the role of Bcpme1 in Botrytis cinerea virulence, a mutant in Bcpme1 was generated by gene disruption. The Bcpme1 mutant showed similar growth on rich medium but reduced growth on pectin medium. Two isozymes of pI 7.4 and 7.1 were detected in pectin liquid-culture supernatants of wild-type strain Bd90 analyzed by isoelectric focusing-polyacrylamide gel electrophoresis, while those of Bcpme1 mutant possessed only the pI 7.1 isozyme. BCPME1, the pI 7.4 isozyme, is the major PME activity, as PME activity is 75% reduced in Bcpme1 mutant. Moreover, the Bcpme1 mutant was less virulent on apple fruits, grapevine, and Arabidopsis thaliana leaves. Those phenotypes were complemented by reintroducing a Bcpme1 copy in the Bcpme1 mutant. These results showed that B. cinerea possessed more than one PME-encoding gene and that BCPME1 is an important determinant of B. cinerea virulence. PMID- 12744466 TI - Morphological correlates of acute and permanent hearing loss during experimental pneumococcal meningitis. AB - In patients with acute bacterial meningitis, hearing loss can be transient but is often permanent. The mechanisms underlying meningitis-associated hearing loss are not fully understood. Therefore, we investigated the morphological correlates of hearing loss in a rat model of pneumococcal meningitis. Transcutaneous intracisternal injection of Streptococcus pneumoniae resulted in a dose-dependent hearing loss (determined by auditory brainstem response audiometry), which was partially reversible during the acute stage. Nevertheless, a severe permanent hearing loss persisted until 2 weeks after infection. Suppurative labyrinthitis was accompanied by blood-labyrinth barrier disruption (determined by cochlear Evans blue extravasation), which correlated closely with hearing loss during the acute stage but not after recovery. Two weeks after infection, spiral ganglion neuronal density was markedly decreased and correlated with the severity of permanent hearing loss. Neuronal loss can be explained by the new finding of meningitis-associated spiral ganglion neuronal necrosis rather than apoptosis (determined by morphology, TUNEL staining, and immunohistochemistry). PMID- 12744467 TI - Vascular patterns in glioblastoma influence clinical outcome and associate with variable expression of angiogenic proteins: evidence for distinct angiogenic subtypes. AB - No data exist on angiogenic patterns and their prognostic impact in human glioblastoma. Such data are relevant for translation of antiangiogenic therapies into clinical applications. Using immunohistochemistry for CD34, we assessed vascular patterns in 114 primary glioblastomas. Vascular patterns comprised unevenly distributed glomeruloid/garland-like/clustered bizarre vascular formations and evenly distributed delicate capillary-like microvessels ("classic" vascular pattern). The combination of low content of bizarre vascular formations and prominent classic vascular pattern (n=29) was an independent factor for longer survival (p=0.006, Cox regression), as well as postoperative high Karnofsky performance status (p=0.005). In patients with a prominent classic vascular pattern, there was no difference of MIB1 labeling index whereas microvessel density and apoptotic index (TUNEL) were significantly higher as compared to all other patients (p<0.05). In addition, diffuse expression of hypoxia-inducible factor (HIF)-1alpha and strong expression of vascular endothelial growth factor were more common (p<0.05, Chi-square test). FISH revealed loss of chromosomes 1p and 19q only in 1/7 long-time survivors with classic pattern. We conclude that vascular patterns in primary glioblastoma influence clinical outcome and associate with variable expression of angiogenic proteins. Our findings denote for the first time distinct angiogenic subtypes of human glioblastoma which may prove relevant for anti-angiogenic therapy approaches. PMID- 12744468 TI - Detection of HIV-1 DNA in microglia/macrophages, astrocytes and neurons isolated from brain tissue with HIV-1 encephalitis by laser capture microdissection. AB - In HIV-1 encephalitis, HIV-1 replicates predominantly in macrophages and microglia. Astrocytes also carry HIV-1, but the infection of oligodendrocytes and neurons is debated. In this study we examined the presence of HIV-1 DNA in different brain cell types in 6 paraffin embedded, archival post-mortem pediatric and adult brain tissues with HIV-1 encephalitis by Laser Capture Microdissection (LCM). Sections from frontal cortex and basal ganglia were stained by immunohistochemistry for CD68 (microglia), GFAP (astrocytes), MAP2 (neurons), and p24 (HIV-1 positive cells) and different cell types were microdissected by LCM. Individual cells or pools of same type of cells were lysed, the cell lysates were subjected to PCR using HIV-1 gag SK38/SK39 primers, and presence of HIV-1 DNA was confirmed by Southern blotting. HIV-1 gag DNA was consistently detected by this procedure in the frontal cortex and basal ganglia in 1 to 20 p24 HIV-1 capsid positive cells, and in pools of 50 to 100 microglia/macrophage cells, 100 to 200 astrocytes, and 100 to 200 neurons in HIV-1 positive cases but not in HIV-1 negative controls. These findings suggest that in addition to microglia, the infection of astrocytes and neurons by HIV-1 may contribute to the development of HIV-1 disease in the brain. PMID- 12744469 TI - Expression of antioxidant enzymes in astrocytic brain tumors. AB - We studied the expression of antioxidant enzymes (AOEs) and related proteins manganese superoxide dismutase (MnSOD), thioredoxin (Trx), thioredoxin reductase (TrxR), and the catalytic (GLCL-c) and regulatory (GLCL-r) subunits of glutamate cysteine ligase (gamma-glutamylcysteinesynthetase) in 433 astrocytomas. Expression of MnSOD was found in 91%, Trx in 46%, TrxR in 66%, GLCL-c 73% and GLCL-r in 89% of the cases. Diffuse astrocytomas showed more intense staining for Trx (p = 0.002), TrxR (p = 0.004), GLCL-c (p = 0.001), GLCL-r (p = 0.04) and MnSOD (p = 0.01) than pilocytic astrocytomas. Within diffuse astrocytomas only Trx (p = 0.0001) and TrxR (p= 0.04) significantly associated with increased malignancy grade. Necrotic tumors were more often immunopositive for Trx (p = 0.001) and TrxR (p = 0.02) and AOE expression was generally higher in mitotically active tumors. Expression of Trx and lack of MnSOD expression was associated with a worse prognosis in diffuse astrocytomas. None of the AOEs had any prognostic value in pilocytic grade I astrocytomas. Familial astrocytomas, which included 23 of the cases studied, did not differ in their expression of MnSOD from sporadic ones. The results show that MnSOD and Trx may influence the biological behaviour of astrocytomas, possibly by modulating cell proliferation and necrosis in these tumors. PMID- 12744470 TI - Soluble factors released by virus specific activated cytotoxic T-lymphocytes induce apoptotic death of astroglioma cell lines. AB - Astrocytomas and astrogliomas represent the most common types of primary tumors in human central nervous system and are associated with high mortality due to the absence of efficient therapy. Here we demonstrate that, upon antigen-specific activation, cytotoxic T-lymphocytes (CTLs) secrete products that inhibit proliferation and induce apoptosis in a significant proportion of astroglioma cell lines. This effect is tumor specific in that normal cultured astrocytes do not develop apoptotic changes upon exposure to supernatant of activated CTLs. Experiments with purified lymphokines and lymphokine specific blocking antibodies indicate that synergistic activities of tumor necrosis factor (TNF)-alpha and interferon (INF)-gamma are required for the apoptosis inducing effect on some astroglioma cell lines. However, this effect appears to be dependent on additional factors produced by activated CTLs. Our results suggest that local application of factors released by activated CTLs or induction of CTL migration and activation in the tumor site may have a therapeutic effect in patients with astrogliomas. PMID- 12744471 TI - Promoter hypermethylation of the DNA repair gene O6-methylguanine-DNA methyltransferase is an independent predictor of shortened progression free survival in patients with low-grade diffuse astrocytomas. AB - The O6-methylguanine-DNA methyltransferase (MGMT) plays a major role in repairing DNA damage from alkylating agents. In several human neoplasms including low-grade diffuse astrocytomas, promoter hypermethylation of MGMT has been shown to correlate with an increased frequency of p53 mutation. In the present study, we analyzed MGMT promoter methylation by the methylation-specific PCR in 49 newly diagnosed WHO grade II astrocytomas and evaluated its clinical usefulness. MGMT promoter methylation was found in 21 (43%) of the 49 tumors. A tight correlation existed between MGMT methylation and p53 protein accumulation (P=0.0424). The presence of MGMT methylation was significantly associated with a shorter progression free survival (PFS) on both univariate analysis (P=0.0014) and multivariate analysis (P=0.0081). It was a more powerful determinant of the PFS than age, sex, performance status, proliferative activity, or p53 expression, and was independent of the extent of surgery. In terms of the overall survival, MGMT methylation demonstrated a prognostic utility in the univariate analysis but not in the multivariate analysis. The present findings indicate that aberrant methylation of the MGMT promoter independently augurs for an unfavorable clinical course in patients with low-grade diffuse astrocytomas. Since the presence of MGMT methylation is expected to predict an increased sensitivity to alkylating chemotherapeutic agents, earlier chemotherapy could serve to improve an unfavorable natural history in tumors with MGMT methylation. PMID- 12744459 TI - Transcriptional changes in response to arbuscular mycorrhiza development in the model plant Medicago truncatula. AB - Significant changes in root morphology and physiology during arbuscular mycorrhiza (AM) development are likely to be controlled by specific gene expression pattern in the host plant. Until now, little was known about transcriptional changes which occur AM-exclusively; that is, they do not occur during other root-microbe associations, nor are they induced by improved phosphate nutrition. In order to identify such AM-exclusive gene inductions of Medicago truncatula, we used a pool of different RNA samples as subtractor population in a suppressive subtractive hybridization (SSH) experiment. This approach resulted in the identification of a number of new AM-regulated genes. None of these genes were expressed in nonmycorrhiza roots or leaves. Electronic data obtained by comparison of the cDNA sequences to expressed sequence tag (EST) sequences from a wide range of cDNA libraries in the M. truncatula EST database (Gene Index, MtGI) support the mycorrhiza specificity of the corresponding genes, because sequences in the MtGI that were found to match the identified SSH-cDNA sequences originated exclusively from AM cDNA libraries. The promoter of one of those genes, MtGst1, showing similarities to plant glutathione-S-transferase (GST) encoding genes, was cloned and used in reporter gene studies. In contrast to studies with the potato GST gene PRP, MtGst 1 promoter activity was detected in all zones of the root cortex colonized by Glomus intraradices, but nowhere else. PMID- 12744472 TI - Granular cell astrocytomas show a high frequency of allelic loss but are not a genetically defined subset. AB - Granular cell astrocytomas (GCA) are an uncommon morphologic variant of infiltrative glioma that contains a prominent population of atypical granular cells. As a rule, they are biologically aggressive compared to similar tumors without granular features. We sought to determine whether GCAs possess distinct genotypic alterations that might reflect their unique morphology or clinical behavior. Eleven GCAs occurring in 7 men and 4 women ranging in age from 46 to 75 years were investigated for genetic alterations of known significance in glial tumorigenesis, including LOH at 1p, 9p, 10q, 17p, and 19q, point mutations of TP53, deletions of p16(CDKN2A) and p14ARF, as well as EGFR amplifications. Tumors included had an infiltrative growth pattern and consisted of large, round cells packed with eosinophilic, PAS-positive granules that varied in quantity, ranging from 30 to 100% of tumor cells. Three tumors were of WHO grade II, one was grade III, and 7 were grade IV lesions. Overall, the tumors showed higher frequencies of LOH at 1p, 9p, 10q, 17p, and 19q than typical infiltrating astrocytomas of similar grades. Losses on 9p and 10q occurred in nearly all cases, including low grade lesions. TP53 mutations were identified in 2 grade IV GCAs, while combined p14ARF and p16(CDKN2A) homozygous deletions were noted in only one grade IV lesion. None showed EGFR amplification. We found no genetic alterations specific for GCA. Instead, it appears that granular cell change occurs across genetic subsets. The high frequency of allelic loss, especially on 9p and 10q, may confer aggressive growth potential and be related to their rapid clinical progression. PMID- 12744475 TI - Peripheral neuropathies in HIV-infected patients in the era of HAART. PMID- 12744476 TI - Human brain parenchymal microglia express CD14 and CD45 and are productively infected by HIV-1 in HIV-1 encephalitis. PMID- 12744474 TI - Regulated expression of sodium-dependent glutamate transporters and synthetase: a neuroprotective role for activated microglia and macrophages in HIV infection? AB - It is now widely accepted that neuronal damage in HIV infection results mainly from microglial activation and involves apoptosis, oxidative stress and glutamate mediated neurotoxicity. Glutamate toxicity acts via 2 distinct pathways: an excitotoxic one in which glutamate receptors are hyperactivated, and an oxidative one in which cystine uptake is inhibited, resulting in glutathione depletion and oxidative stress. A number of studies show that astrocytes normally take up glutamate, keeping extracellular glutamate concentration low in the brain and preventing excitotoxicity. This action is inhibited in HIV infection, probably due to the effects of inflammatory mediators and viral proteins. Other in vitro studies as well as in vivo experiments in rodents following mechanical stimulation, show that activated microglia and brain macrophages express high affinity glutamate transporters. These data have been confirmed in chronic inflammation of the brain, particularly in SIV infection, where activated microglia and brain macrophages also express glutamine synthetase. Recent studies in humans with HIV infection show that activated microglia and brain macrophages express the glutamate transporter EAAT-1 and that expression varies according to the disease stage. This suggests that, besides their recognized neurotoxic properties in HIV infection, these cells also have a neuroprotective function, and may partly make up for the inhibited astrocytic function, at least temporarily. This hypothesis might explain the discrepancy between microglial activation which occurs early in the disease, and neuronal apoptosis and neuronal loss which is a late event. In this review article, we discuss the possible neuroprotective and neurotrophic roles of activated microglia and macrophages that may be generated by the expression of high affinity glutamate transporters and glutamine synthetase, 2 major effectors of glial glutamate metabolism, and the implications for HIV-induced neuronal dysfunction, the underlying cause of HIV dementia. PMID- 12744477 TI - October 2002: 27-year-old female with epilepsy. AB - The October 2002 Case of the Month (COM). The patient was a 27-year-old woman with a history of partial complex seizures at age 7. At age 20 her seizures changed in character and became progressively worse. Neuroimaging studies showed atrophy of the right hemisphere and contralateral cerebellar atrophy. Following a biopsy, she was scheduled for a surgical procedure, but unfortunately she expired at home during her sleep a week later. Examination of the brain confirmed the hemi-atrophy of the right cerebral hemisphere and left cerebellum. Microscopic examination showed severe gliosis and perivascular lymphocytic infiltrates in many areas. A diagnosis of Rasmussen's encephalitis was made. Rasmussen's encephalitis is a chronic neurological disorder, first described in 1958. The active neurological decline lasts from 1 to 20 years and the patients then remain stable with a fixed neurological deficit and residual seizures. Pathological examination shows a chronic encephalitis confined to one hemisphere. In the active phase, neuronophagia, activated microglial cells (rod cells), microglial nodules, and perivascular lymphocytic infiltrates, are present. In the more chronic phase neuronal loss and gliosis predominate. The etiology of Rasmussen's encephalitis is unknown but viral infection and autoimmunity have been implicated. The treatment of choice is functionally complete hemispherectomy with complete disconnection of the frontal and occipital lobes. PMID- 12744473 TI - Changing patterns in the neuropathogenesis of HIV during the HAART era. AB - Rapid progress in the development of highly active antiretroviral therapy has changed the observed patterns in HIV encephalitis and AIDS-related CNS opportunistic infections. Early in the AIDS epidemic, autopsy studies pointed to a high prevalence of these conditions. With the advent of nucleoside reverse transcriptase inhibitors, the prevalence at autopsy of opportunistic infections, such as toxoplasmosis and progressive multifocal leukoencephalopathy, declined while that of HIV encephalitis increased. After the introduction of protease inhibitors, a decline in both HIV encephalitis and CNS opportunistic infections was observed. However, with the increasing resistance of HIV strains to antiretrovirals, there has been a resurgence in the frequency of HIV encephalitis and HIV leukoencephalopathy. HIV leukoencephalopathy in AIDS patients failing highly active antiretroviral therapy is characterized by massive infiltration of HIV infected monocytes/macrophages into the brain and extensive white matter destruction. This condition may be attributable to interactions of anti retrovirals with cerebrovascular endothelium, astroglial cells and white matter of the brain. These interactions may lead to cerebral ischemia, increased blood brain barrier permeability and demyelination. Potential mechanisms of such interactions include alterations in host cell signaling that may result in trophic factor dysregulation and mitochondrial injury. We conclude that despite the initial success of combined anti-retroviral therapy, more severe forms of HIV encephalitis appear to be emerging as the epidemic matures. Factors that may contribute to this worsening include the prolonged survival of HIV-infected patients, thereby prolonging the brain's exposure to HIV virions and proteins, the use of increasingly toxic combinations of poorly penetrating drugs in highly antiretroviral-experienced AIDS patients, and selection of more virulent HIV strains with higher replication rates and greater virulence in neural tissues. PMID- 12744478 TI - November 2002: a 72-year-old woman with a pineal gland mass. AB - The November 2002 COM. A 72-year-old immunocompetent woman presented with recent confusion, memory loss, visual and a gait disturbance. MRI scans demonstrated a T2 hypointense, gadolinium enhancing mass in her pineal region. A whole body CT scan identified a large retroperitoneal mass and significant lymphadenopathy. She underwent a biopsy of the pineal lesion. The intraoperative smear contained discohesive, basophilic malignant cells of moderate size that had only scant cytoplasm, an increased nuclear-to-cytoplasmic ratio and irregular, hyperchromatic nuclei containing prominent nucleoli. Mitotic figures, apoptotic and lymphoglandular bodies were frequent. Histologic examination showed malignant cells infiltrating and almost completely replacing the residual pineal gland. The tumor cells immunoreacted with LCA and CD-20. Scattered CD3 and UCHL-1 positive cells were also present. The diagnosis was stage IV, high grade, large B-cell lymphoma involving the pineal gland and retroperitoneum. Four cycles of systemic chemotherapy significantly reduced the patient's retroperitoneal mass. However, a repeat head MRI scan continued to show gadolinium-enhancement in the pineal region, now with extension into contiguous brain parenchyma and the subependymal region. The patient died 4 months after presentation. At autopsy, the lymphoma encased the pineal region and disseminated along the ventricular walls and into the subarachnoid spread. Pineal-region tumors are rare in the elderly population, and although unusual, lymphomas should be considered in this context. The behavior of this circumventricular organ lymphoma was comparable to both peripheral and primary central nervous system lymphomas. PMID- 12744479 TI - December 2002: 19-year old male with febrile illness after jet ski accident. AB - The December 2002 COM. A 19-year-old healthy male fell into stagnant water of the intercostal waterway (salt water of South Florida), following a jet ski accident. He sustained minor superficial injuries but engulfed significant quantities of water and sediment. A few days later he developed bifrontal headaches, vomiting, a stiff neck and a temperature of 102 degrees F. A CT scan on admission without contrast was negative. The CSF had markedly elevated white count but bacterial and fungal cultures were negative. He became progressively lethargic. On the fifth day he developed seizure activity. He expired the next day despite antibiotics. Gross examination of the brain at autopsy revealed edema, cerebellar tonsillar herniation and purulent meningitis. Microscopic examination revealed a massive leptomeningeal inflammatory infiltrate composed of neutrophils, lymphocytes, and numerous histiocyte-like cells. The inflammatory infiltrate extended into the cerebral parenchyma in numerous areas also involving the cerebellum, brainstem and ventricular system. Given the exposure to stagnant water (later confirmed to be a man-made fresh water lake), and the numerous histiocytic-like cells, suspicion for an amebic etiology of the disease process was raised and the CDC identified the ameba as Naegleria Fowleri. Infection by Naegleria Fowleri, a free-living ameba, occurs after exposure to polluted water in man-made fresh water lakes, ponds, swimming pools, particularly during the warm weather months when the thermophilic ameba grows well. The pathologic substrate of the infection is an acute hemorrhagic, necrotizing meningo encephalitis mainly at the base of the brain, brainstem and cerebellum occurring in young, healthy individuals. PMID- 12744481 TI - Intraoperative detection of traumatic coagulopathy using the activated coagulation time. AB - Traumatic coagulopathy manifests as a hypocoagulable state associated with hypothermia, acidosis, and coagulation factor dilution. The diagnosis must be made clinically because traditional coagulation tests are neither sensitive nor specific and take too long to be used for intraoperative monitoring. We hypothesized that the activated coagulation time (ACT) would reflect the global coagulation status of traumatized patients and would become elevated as coagulation reserves become exhausted. A prospective protocol was used to study 31 victims of major trauma who underwent immediate surgical Intervention. Victims of major head trauma were excluded and patients were selected at random over an 8 month period. At least two serial intraoperative blood samples were obtained at 15-min intervals via indwelling arterial catheters. A Hemochron model 801 coagulation monitor was used to measure the ACT. Of the 31 patients studied, 7 became clinically coagulopathic and 24 did not. The ACT measurements of coagulopathic and noncoagulopathic trauma patients were significantly different by multiple statistical comparisons. Both groups differed from normal, nontraumatized patients. The coagulopathic trauma patients had significantly elevated values when compared with other trauma patients or to normal values. We conclude that a low ACT reflects the initial hypercoagulability associated with major trauma and an elevated ACT is an objective indicator that the coagulation system reserve is near exhaustion. An elevated ACT may represent an indication for considering damage control maneuvers or more aggressive resuscitation. PMID- 12744480 TI - Recombinant human growth hormone increases thyroid hormone-binding sites in recovering severely burned children. AB - Thyroxine (T4), Tri-iodothyronine (T3), and total serum protein levels are reduced in severely burned children. T4 and T3 are carried on serum transport proteins via thyroid hormone-binding sites (THBS). Treatment of bums with recombinant human growth hormone (rhGH) increases albumin (Alb) and prealbumin (PreAlb), which bind nearly 30% of circulating T4 and T3. This study investigated the effect of rhGH on THBS sites in burned children. Records of 11 acutely burned children with a total body surface area burned >40% who were randomized to either a daily subcutaneous injection of rhGH at 0.05 mg/kg/day or placebo for 6 months after discharge from hospital were reviewed. Thyroxine uptake percentage (TU%), Total T4 levels (TT4), free thyroxine index, Alb and PreAlb, and height and weight measurements taken at discharge and 6 months later were compared in both groups. In the six children who received rhGH, mean TU% had decreased from 41 +/- 1 to 33 +/- 1% by 6 months postdischarge, (P < 0.001), mean TT4 increased from 5.8 +/- 0.3 to 8.1 +/- 0.8 microg/dL, (P < 0.02), mean Alb increased from 2.0 +/- (0.6) to 3.5 +/- (0.1) g/dL (P < 0.0001), and mean PreAlb increased from 8.7 +/- 0.7 to 16.5 +/- 2.1 mg/dL, (P < 0.006). There were no significant changes in the five children in the placebo (control group), and height and weight did not significantly change in either group. rhGH significantly increases THBS in severely burned children, possibly through increases in serum Alb and PreAlb. The increases in circulating thyroxine observed in this group may be involved in the attenuation of growth arrest. PMID- 12744482 TI - Antibiotics improve survival and alter the inflammatory profile in a murine model of sepsis from Pseudomonas aeruginosa pneumonia. AB - Differing antibiotic regimens can influence both survival and the inflammatory state in sepsis. We investigated whether the addition and/or type of antimicrobial agent could effect mortality in a murine model of Pseudomonas aeruginosa pneumonia-induced sepsis and if antibiotics altered systemic levels of cytokines. FVB/N mice were subjected to intratracheal injection of pathogenic bacteria and were given gentamicin, imipenem, or 0.9% NaCl 2 h after surgery, which continued every 12 h for a total of six doses. Survival at 7 days (n = 24 in each group) was 100% for mice given gentamicin, 88% for mice given imipenem, and 8% for sham mice treated with 0.9% NaCl (P < 0.0001). Systemic interleukin (IL) 6 levels were assayed 6 h postoperatively on all mice to see if they were predictive of outcome. Plasma IL-6 levels above 3,600 pg/mL were associated with a 100% mortality, levels under 1,200 pg/mL were associated with a 100% survival, and levels between 1,200 and 3,600 pg/mL had no utility in predicting mortality. In a separate experiment, mice were sacrificed at 3, 6, 12 or 24 h after instillation of P. aeruginosa and were assayed for levels of TNF-alpha, IL-6, IL 10, and IL-12. Significant alterations in the proinflammatory cytokines TNF-alpha and IL-6 were present at all time points except 3 h between mice treated with antibiotics and sham controls. In contrast, statistically significant differences in the anti-inflammatory cytokine IL-10 were present between the groups only at 6 h, and levels of IL-12 were similar at all time points. These results indicate that both gentamicin and imipenem increase survival at least 10-fold in a model of pneumonia-induced monomicrobial sepsis, and this is predominantly associated with a down-regulation of proinflammatory cytokines. PMID- 12744484 TI - Hemorrhagic shock induces an S 100 B increase associated with shock severity. AB - S 100 B is a glial marker of cerebral Injury. In a previous clinical study, we found an S 100 B increase within the first 24 h in patients with multiple trauma and hemorrhagic shock but without cerebral trauma. The aim of our current experimental study was to determine whether this posttraumatic S 100 B increase is caused by extracerebral soft tissue injury or by hemorrhagic shock and whether it is associated with the severity of hemorrhagic shock. Hemorrhagic shock was achieved by bleeding anesthetized rats to a mean arterial pressure (MAP) of 30-35 mmHg through a femoral catheter and maintaining this MAP until incipient decompensation. At incipient decompensation, MAP was either increased immediately to 40-45 mmHg (moderate shock) or was maintained until 40% of shed blood had been returned (severe shock), and then increased to 40-45 mmHg. Resuscitation was provided after 40-45 mmHg MAP had been maintained for 40 min. Soft tissue injury was achieved by midline laparotomy performed at the onset of hemorrhagic shock or without shock and was maintained for 30 min. Hemorrhagic shock caused an early S 100 B increase at the onset of decompensation. S 100 B remained increased for 24 h and was significantly higher after severe than after moderate shock. In contrast, soft tissue injury without hemorrhagic shock caused no S 100 B increase. The data presented demonstrate for the first time that the S 100 B increase is induced by hemorrhagic shock and is associated with the severity of shock. PMID- 12744483 TI - Systemic and hepatosplanchnic hemodynamic and metabolic effects of the PARP inhibitor PJ34 during hyperdynamic porcine endotoxemia. AB - Activation of the poly(ADP-ribose)polymerase (PARP), a highly energy-consuming DNA-repairing enzyme, plays a crucial role in the pathogenesis of multiorgan failure. Most results, however, were derived from experiments with hypodynamic shock states characterized by a markedly decreased cardiac output (CO) and/or using a pretreatment approach. Therefore, we investigated the effects of the novel potent and selective PARP-1 inhibitor PJ34 in a posttreatment model of long term, volume-resuscitated porcine endotoxemia. Anesthetized, mechanically ventilated and instrumented pigs received continuous intravenous (i.v.) lipopolysaccharide (LPS) over 24 h. Hydroxyethyl starch was administered to maintain a mean arterial pressure > 65 mmHg. After 12 h of LPS infusion, the animals were randomized to receive either vehicle (Control, n = 9) or i.v. PJ34 (n = 6; 10 mg/kg over 1 h followed by 2 mg/kg/h until the end of the experiment). Measurements were performed before as well as at 12, 18, and 24 h of LPS infusion. In all animals CO increased because of reduced systemic vascular resistance (SVR) and fluid resuscitation. PJ34 further raised CO (P < 0.05 vs. control group) as the result of a higher stroke volume indicating its positive inotropic effect. In addition, it diminished the rise in the ileal mucosal arterial PCO2 gap, which returned to baseline levels at 24 h of LPS, and improved the gut lactate balance (P = 0.093 PJ34 vs. control) together with significantly lower portal venous lactate/pyruvate ratios. By contrast, it failed to influence the LPS-induced derangements of liver metabolism. Incomplete PARP inhibition because of dilutional effects and/or an only partial efficacy when used in post treatment approaches may account for this finding. PMID- 12744485 TI - Neuroprotective effects of coenzyme Q10 at rostral ventrolateral medulla against fatality during experimental endotoxemia in the rat. AB - Coenzyme Q10 (CoQ10, ubiquinone) is a highly mobile electron carrier in the mitochondrial respiratory chain that also acts as an antioxidant. We evaluated the neuroprotective efficacy of CoQ10 against fatality in an experimental model of endotoxemia that mimics systemic inflammatory response syndrome using a novel water-soluble formulation of this quinone derivative. Experiments were conducted in adult male Sprague-Dawley rats that were maintained under propofol anesthesia. Intravenous administration of Escherichia coli lipopolysaccharide (LPS; 30 mg/kg) induced progressive hypotension, with death ensuing within 4 h. The sequence of cardiovascular events during this LPS-induced endotoxemia can be divided into a reduction (Phase I), followed by an augmentation (Phase II; "pro-life" phase) and a secondary decrease (Phase III; "pro-death" phase) in the power density of the vasomotor components (0-0.8 Hz) of systemic arterial pressure signals. Pretreatment by microinjection bilaterally of CoQ10 (1 or 2 microg) into the rostral ventrolateral medulla (RVLM), the medullary origin of sympathetic vasomotor tone, significantly diminished mortality, prolonged survival time, and reduced the slope or magnitude of the LPS-induced hypotension. CoQ10 pretreatment also significantly prolonged the duration of and augmented the total power density of the vasomotor components of systemic arterial pressure signals in Phase II endotoxemia. The increase in superoxide anion production induced by LPS at the RVLM during Phases II and III endotoxemia was also significantly blunted. We conclude that CoQ10 provides neuroprotection against fatality during experimental endotoxemia by reducing superoxide anion production at the RVLM, whose neuronal activity is intimately related to the "life-and-death" process. PMID- 12744486 TI - Endotoxin-induced endothelial cell proinflammatory phenotypic differentiation requires stress fiber polymerization. AB - Endotoxin-induced intercellular adhesion molecule-1 (ICAM-1) and interleukin 8 (IL-8) production in endothelial cells, which is mediated by Toll-receptor signaling, is essential for optimal neutrophil recruitment and migration during sepsis. Endotoxin also causes stress fiber polymerization that has recently been shown to affect intracellular signaling. However, the role of this polymerization process on endothelial-induced neutrophil adhesion and migration is unknown. Human umbilical vein endothelial cells (HUVEC) were stimulated with lipopolysaccharide (LPS). Selected cells were pretreated with cytochalasin D (CD) or lactrunculin A (LA), agents that disrupt actin polymerization. Cellular protein was extracted and analyzed by Westem blot for the phosphorylated form of IL-1-associated kinase (IRAK) and production of ICAM-1. Extracted nuclear protein was analyzed by Western blot and electrophoretic mobility shift assay (EMSA) for nuclear translocation and activity of NF-kappaB. IL-8 production was determined by enzyme-linked immunoabsorbant assay (ELISA). Neutrophil adhesion was assayed fluorometrically using calcein-AM-labeled neutrophils on treated endothelial cells. LPS treatment led to phosphorylation of IRAK, and subsequent NF-kappaB translocation and activation. This cellular signaling was followed by ICAM-1 expression and IL-8 production. Pretreatment of cells with CD or LA led to a significant inhibition of IRAK phosphorylation, and NF-kappaB nuclear translocation and activation. Actin depolymerization also significantly inhibited LPS-induced ICAM-1 and IL-8 production. HUVEC pretreated with CD or LA demonstrated significant inhibition of LPS-induced neutrophil adhesion. Endotoxin induced actin polymerization is essential for optimal intracellular signaling through IRAK and NF-kappaB. Failure of these signaling events is associated with a marked reduction in adhesion molecule production, IL-8 production, and neutrophil adhesion. These findings support the necessity of stress fiber polymerization for optimal recruitment of neutrophils during sepsis. PMID- 12744487 TI - Reduced blood-to-tissue albumin movement after plasmapheresis. AB - We tested the hypothesis that a decrease in the blood-to-tissue movement of albumin contributes to the recovery of plasma albumin and plasma volume after acute plasma protein depletion (plasmapheresis). Awake and unrestrained male Sprague-Dawley rats (220-320 g) fitted with jugular catheters were plasmapheresed, and plasma volume, plasma albumin, and total plasma protein content were measured at 1, 5, 24, and 48 h postplasmapheresis. Plasma volume recovered to baseline within 1 h (4.6 +/- 0.42 vs. 4.7 +/- 0.46 mL/100 g body weight (bw), remained at baseline from 5 h to 24 h but increased to 5.5 + 0.57 mL/100 g bw at 48 h (P < 0.05). Plasma albumin and total protein content recovered rapidly but remained below baseline levels at 1 h (10.05 +/- 0.98 vs. 12.33 +/- 1.29 and 19.75 +/- 1.75 vs. 24.73 +/- 2.56 mg/100 g bw, respectively). Plasma protein content retumed to baseline by 5 h of recovery. Tissue uptake of I125-labeled albumin decreased in the heart, skin, skeletal muscle, and small Intestines of plasmapheresed rats (P < 0.05). These data support the hypothesis that a reduction in albumin efflux from the vascular space contrlbutes to the recovery of plasma albumin and total protein content during plasma volume recovery and eventual expansion after plasmapheresis. PMID- 12744488 TI - Enalapril does not alter adhesion molecule levels in human endotoxemia. AB - The angiotensin-converting enzyme inhibitor (ACE-I) enalapril has been shown to lower elevated levels of circulating adhesion molecules (cAM) in critically ill patients. To delineate the mechanisms of this possibly beneficial effect of enalapril, we studied the acute effects of enalapril in a well-defined model of endotoxin-triggered, cytokine-mediated cAM up-regulation. In a randomized, controlled trial, 30 healthy male volunteers received 2 ng/kg lipopolysaccharide (LPS) after pretreatment with placebo or 20 mg/day enalapril for 5 days or with a single dose of 20 mg of enalapril 2 h before LPS infusion. LPS infusion increased TNF levels 300-fold above normal, circulating (c) E-selectin levels by 425% (CI, 359%-492%), and P-selectin, VCAM-1, ICAM-1, and von Willebrand factor levels by 47%-74%. LPS infusion also enhanced ICAM-1 and CD11b expression 2- to 3-fold on monocytes. However, no differences were seen between treatment groups (P > 0.05), despite 95% inhibition of ACE activity by enalapril. Inhibition of ACE activity by enalapril does not influence plasma indices of endothelial activation after endotoxin infusion in healthy individuals. Our results do not support the concept of a beneficial clinical effect of enalaprilat in septicemia. PMID- 12744489 TI - Serine proteases are involved in the pathogenesis of trauma-hemorrhagic shock induced gut and lung injury. AB - The objective of this work was to test the hypothesis that Intraluminal serine proteases are involved in trauma-hemorrhagic shock (T/HS)-induced intestinal and lung injury. Male Sprague-Dawley rats were administrated the serine protease inhibitor (6-amidino-2-naphthyl p-guanidinobenzoate dimethanesulfate, Nafamostat) either intraluminally into the gut or intravenously after a laparotomy (trauma) and then subjected to 90 min of hemorrhagic shock (T/HS) or sham shock (T/SS). Intestinal and lung injury was assessed at 3 h after resuscitation with Ringer's lactate solution. In a second set of experiments, mesenteric lymph was collected from the groups of rats subjected to T/HS or T/SS and its ability to activate normal neutrophils was tested. Lung permeability, pulmonary myeloperoxidase levels, and the bronchoalveolar lavage fluid protein to plasma protein ratio were increased after T/HS but were significantly decreased in the T/HS rats receiving intraluminal (P < 0.05), but not intravenous, nafamostat. Likewise, T/HS-induced intestinal villus injury was less in the nafamostat-treated shock rats (P < 0.05). Last, the ability of T/HS mesenteric lymph to increase PMN CD11b expression or prime neutrophils for an augmented respiratory burst was significantly reduced by the intraluminal administration of nafamostat. Because intraluminal nafamostat reduced T/HS-induced gut and lung injury as well as the neutrophil activating ability of intestinal T/HS lymph, the presence of serine proteases in the ischemic gut may play an important role in T/HS-induced gut and hence lung injury. PMID- 12744491 TI - Pharmacological preconditioning protects lung injury induced by intestinal ischemia/reperfusion in rat. AB - Intestinal ischemia/reperfusion (IIR) is a critical and triggering event in the development of distal organ dysfunction, frequently involving the lungs. Respiratory failure is a common cause of death and complications after intestinal I/R. Stress protein heme oxygenase-1 (HO-1) confers the protection against a variety of oxidant-induced cell and tissue injuries. The aim of this study was to investigate the hypothesis that the induced HO-1 expression by pharmacological preconditioning with anticancer drug doxorubicin (Dox) could protect the lung injury induced by intestinal I/R. Intravenous administration of Dox induced HO-1 expression in the lungs and high levels of the expression were sustained at least to 48 h after the injection. Therefore, as pharmacological preconditioning, a low dose of Dox was injected intravenously into rats at 48 h before the start of intestinal ischemia. Rats underwent intestinal I/R by superior mesenteric artery occlusion for 120 min followed by 120 min of reperfusion. Preconditioning with Dox significantly ameliorated the lung injury induced by the intestinal I/R. Administration of a specific inhibitor of HO activity reduced the efficacy of the preconditioning. Our results suggest that this improvement may be mediated at least in part by the HO-1 induction. These findings may offer interesting perspectives for patient management In Intestinal surgical operation and intestine transplantation. PMID- 12744490 TI - Attenuation of leukocyte adhesion by recombinant TNF-binding protein after hemorrhagic shock in the rat. AB - Ischemia/reperfusion injury involves a large number of humoral and cellular mediators that activate leukocytes that subsequently migrate to local tissues. Tumor necrosis factor (TNF)-alpha may be one of the most important mediators of this post-shock inflammatory response. In this study, we investigated the influence of a recombinant Type I (55 kDa) TNF-binding protein (TNF-BP) on leukocyte-endothelial interactions in the liver after hemorrhagic shock. Hemorrhagic shock was induced in female Sprague-Dawley rats (40 mmHg for 90 min) and a standardized resuscitation regimen was applied. At the time of resuscitation, animals were treated intravenously with either TNF-BP 4 mg/kg or placebo. The liver microcirculation was investigated using intravital fluorescence microscopy and immunohistochemistry at 5 h and 48 h after reperfusion. At 5 h, treatment with TNF-BP significantly reduced temporary leukocyte adhesion in the liver sinusoids as well as mean adhesion time of leukocytes in the hepatic central vein. In contrast, after 48 h, permanent leukocyte adhesion in the central hepatic vein was significantly reduced in the group receiving TNF-BP, whereas temporary leukocyte adhesion and mean adhesion time did not differ between the two groups. Both types of leukocyte adhesion, rolling adhesion after 5 h and firm adhesion after 48 h, were reduced in the group treated with TNF-BP, thereby suggesting a long-lasting anti-inflammatory effect. PMID- 12744492 TI - Lipopolysaccharide and cecal ligation/puncture differentially affect the subcellular distribution of the pregnane X receptor but consistently cause suppression of its target genes CYP3A. AB - The repressed expression of cytochrome P450 (CYP) enzymes in septic patients contributes significantly to therapeutic failures. Mice treated with sepsis inducing agent lipopolysaccharide (LPS) sequentially express reduced mRNA levels of the pregnane X receptor (PXR) and its target genes Cyp3a(s), suggesting that reduction of Cyp expression is associated with the repression of PXR. The present study was undertaken to determine whether septic rats induced by LPS and cecal ligation/puncture (CLP) express reduced levels of rat PXR protein and whether the subcellular distribution of PXR is altered in septic conditions. Rats were treated with LPS (55 vs. 1 mg/kg) or underwent CLP, and the expression of CYP3A and PXR was determined. In LPS-treated rats, the expression of CYP3A enzymes was consistently decreased regardless of the doses used. In contrast, high dose and repeated low dose of LPS caused significant decreases on the nuclear PXR, whereas the opposite was true with the cytosolic PXR. When rats were administered with only a single low dose of LPS, both nuclear and cytosolic PXR levels were significantly increased. In the CLP model, rats undergoing CLP for 30 h expressed significantly lower levels of CYP3A but the PXR levels were not significantly altered. In addition, when rats were treated with dexamethasone, a significant induction of CYP3A was detected. However, such an induction was markedly antagonized by the treatment with LPS. The differential changes on the levels of the nuclear PXR and CYP3A between LPS and CLP models suggest that PXR plays negligible roles in the constitutive expression of CYP3A. The antagonism of LPS against dexamethasone-mediated CYP3A induction suggests that endotoxemia minimizes the inducibility of PXR target genes. PMID- 12744493 TI - Role of p38 mitogen-activated protein kinase in lung injury after burn trauma. AB - This study was undertaken to evaluate the effect of SB203580, a specific p38 mitogen-activated protein (MAP) kinase inhibitor, on burn-induced lung injury as well as the release of tumor necrosis factor (TNF)-alpha and interleukin (IL) 1beta in rats to characterize the role of p38 MAP kinase in lung injury after burn trauma. Sprague-Dawley rats were divided into three groups: 1) sham group, or rats who underwent sham burn; 2) control group, or rats given third-degree burns over 30% total body surface area (TBSA) and lactated Ringer solution for resuscitation; and 3) SB203580 group, or rats given burn injury and lactated Ringers solution with SB203580 inside for resuscitation. Pulmonary injury was assessed at 24 h by pulmonary capillary permeability determined with fluorescein isothiocyanate-labeled albumin and lung histologic analysis. TNF-alpha and IL 1beta protein in bronchoalveolar lavage fluid and serum were measured by enzyme linked immunosorbent assay and p38 MAP kinase was activity determined in lung by Western blot analysis. These studies showed that significant activation of p38 MAP kinase at 24 h postburn compared with control. Burn trauma resulted in increased pulmonary capillary leakage permeability, elevated levels of TNF-alpha and IL-1beta in bronchoalveolar lavage fluid and serum, and worsened histologic condition. SB203580 inhibited the activation of p38 MAP kinase, reduced the levels of TNF-alpha and IL-1beta, and prevented burn-mediated lung injury. These data suggest that p38 MAP kinase activation is one important aspect of the signaling event that may mediate the release of TNF-alpha and IL-1beta and contributes to burn-induced lung injury. PMID- 12744494 TI - The effect of an intestinal ischemia-reperfusion injury on renal nerve activity among rats. AB - An intestinal ischemia-reperfusion injury (IIR) may induce renal tubular dysfunction and a reduction in renal blood flow that may be related to the alteration of renal-nerve activity. A rat model of IIR injury was established. The superior mesenteric artery was clamped for 120 min, constituting the ischemic period, and was then released for 60 min, thus constituting the reperfusion period. Renal-nerve activity, renal function, and hemodynamic changes were recorded during the different periods. The levels of calcitonin gene-related peptide (CGRP) in portal-vein blood and intestinal tissue were investigated here. In the reperfusion period, the efferent renal-nerve activity (ERNA) was markedly elevated (94.3% +/- 21.6% higher than the baseline value), such an elevation being only partially reversed by fluid expansion (29.3% +/- 5.2% higher than the baseline value). The elevation of ERNA contributed to the renal blood-flow reduction from 6.8 +/- 0.3 mL/min/g to 2.0 +/- 0.4 mL/min/g, and decreased diuretic and natriuretic responses. The afferent renal nerve activity (ARNA) was markedly depressed (45.7% +/- 8.1% lower than the baseline value) during the reperfusion period. This depression was not reversed by fluid expansion, suggesting that the baroreflex was not responsible for this effect. The blunted ARNA also contributed to the elevation of ERNA by way of a renorenal reflex. The potent vasodilator neuropeptide in the gut, CGRP, revealed an increased level in the portal-vein blood (92.2 +/- 4.4 pg/mL vs. 57.8 +/- 0.6 pg/mL) and also in intestinal tissue (655.8 +/- 115.9 pg/mL vs. 60.5 +/- 9.4 pg/mL) with a time matched related pattern with the change to renal-nerve activity, suggesting CGRP's role regarding changes in renal-nerve activity. This study indicates that the elevated ERNA level associated with IIR injury is related to a systemic hypotension-induced baroreflex, the contra-lateral inhibition of ARNA, and possibly also gut-released CGRP. In regards to an IIR injury, the depressed ARNA reflects the involvement of a renal sensory- impairment mechanism. PMID- 12744496 TI - Appendiceal imaging. PMID- 12744495 TI - Superoxide production in the vasculature of lipopolysaccharide-treated rats and pigs. AB - Sepsis is associated with increased production of reactive oxygen species (ROS); however, the metabolic sources of increased ROS are not well understood. We hypothesized that the recently described nonphagocytic NAD(P)H oxidase system could be an important source of the ROS superoxide anion (O2-) during sepsis, and the interaction of O2- with nitric oxide (NO) may contribute to sepsis-induced vascular Injury. To evaluate this issue, we measured O2- production before and after treatment with lipopolysaccharide (LPS) in rats, who are Inducible NO synthase producers (NOSII) and in pigs, who do not produce NOSII. LPS increased O2- production in aorta from rats from 0.38 +/- 0.07 nmol/mg/10 min to 1.18 +/- 0.23 nmol/mg/10 min, (P = 0.001) in rats, and 0.63 +/- 0.05 nmol/mg/10 min to 1.5 +/- 1.6 nmol/mg/10 min (P = 0.001) in carotid arteries from pigs. Components of NAD(P)H oxidase, including p22(phox), gp91(phox), p47(phox), p67(phox), mRNA and p22(phox), and gp91(phox) proteins were present in rat aorta and aorta and carotid arteries from pigs. Expression mildly increased in rats, but not in pigs. In rats, NADH and NADPH greatly increased O2- production with no difference in untreated versus LPS-treated rats. The addition of L-NAME increased NADH dependant O2- production from 75 +/- 3 nmol/O2-/mg/10 min to 113 +/- 7 nmoVO2 /mg/10 min in LPS-treated rats, but had no effect in untreated rats. In pigs, the NADH-stimulated O2- production was 43 +/- 8 nmol/mg/10 min before and 63 +/- 4.3 nmol/mg/10 min after LPS even without L-NAME (P < 0.05). In contrast to LPS treated rats, L-NAME markedly decreased NADH-stimulated O2- production (63 +/- 4 nmol/mg/10 min to 33 +/- 5.6 nmol/mg/10 min, P < 0.01). Luminol-enhanced chemiluminescence was also Increased in porcine carotid arteries after LPS treatment, which is consistent with peroxynitrite formation. Our results indicate that components of NAD(P)H oxidase are present in vessels of pigs and rats and there is substantial NADH-dependent O2- production that is increased after LPS. However, the behavior of NAD(P)H oxidase in NOSII-producing and nonproducing species differs with a reduction of O2- by NO in rats and NO-dependent production in pigs. PMID- 12744497 TI - Surgical evaluation of appendicitis in the new era of radiographic imaging. AB - Appendicitis remains a common disease with over 250,000 cases in the United States per year. Surgical intervention has been the primary treatment for over a century, and clinical evaluation has been the mainstay for diagnosis of acute appendicitis for decades. Appendiceal imaging modalities have become increasingly popular to assist with the diagnosis of appendicitis, and the surgeon is no longer the sole diagnostician for this disease. Radiologists and emergency medical physicians have entered the arena. This review examines the surgical and radiological literature to evaluate the current atmosphere of how surgeons, radiologists, and emergency physicians interact in the diagnosis of appendicitis. PMID- 12744498 TI - Ultrasonography and computed tomography in suspected acute appendicitis. AB - Patients with typical signs and symptoms of acute appendicitis should have a prompt surgical consultation for timely appendectomy. The use of diagnostic imaging tests such as CT scan or ultrasonography should be selective in those with atypical presentation or findings. The high accuracy of imaging tests can be achieved when they are utilized according to well thought out clinical pathways. The use of imaging modalities in suspected acute appendicitis should be complement to, but not replacing, clinical assessment and judgement. The skillful use of clinical assessment and imaging modalities will reduce the negative appendectomy rate yet assure low perforation and mortality rates. PMID- 12744499 TI - Imaging of acute appendicitis in children. AB - Acute appendicitis is the most common acute abdominal condition that results in surgical intervention in childhood. The clinical diagnosis of acute appendicitis in children can be challenging. Approximately one-third of children with the condition have atypical clinical findings and are initially managed nonoperatively. Complications associated with delayed diagnosis of this condition include perforation, abscess formation, peritonitis, sepsis, bowel obstruction, infertility, and death. The use of cross sectional imaging has proven useful for the evaluation of suspected acute appendicitis in children. Both graded compression sonography and CT have been widely utilized in the imaging assessment of the condition. The principal advantages of sonography are its lower cost, lack of ionizing radiation, and ability to assess ovarian pathology that can often mimic acute appendicitis in female patients. The principal advantages of CT include less operator dependency than sonography as reflected by a higher diagnostic accuracy, and enhanced delineation of disease extent in perforated appendicitis. PMID- 12744500 TI - Sonography of acute appendicitis. AB - Graded compression sonography is an established imaging modality in the clinical setting of acute right lower quadrant pain or to diagnose acute appendicitis because of its easy assessability, noninvasiveness, real-time imaging. However, the ability to accurately diagnose appendicitis can be affected by several factors including operator dependence, deeper-located appendix, and obesity or muscularity of the patient. However, adjuvant techniques utilizing advanced equipment and accumulated operator's experience to conventional graded compression sonography will yield more frequent detection of the vermiform appendix and more accurate results of acute appendicitis on sonography. This article introduces adjuvant techniques and various know-how of real field in the detection of the vermiform appendix and diagnosis of acute appendicitis. PMID- 12744502 TI - CT imaging in acute appendicitis: techniques and controversies. PMID- 12744501 TI - Appendiceal imaging: which test is best? AB - Radiologic imaging has been clearly shown to be useful for the evaluation of patients clinically suspected of having appendicitis with CT and ultrasound being the most frequently recommended modalities. However, controversy still exists as to which modality is the technique of choice for appendiceal imaging. Our experience has led us to conclude that a standard helical abdominopelvic CT is the initial test of choice for the evaluation of the adult patient suspected of having acute appendicitis. Focused appendiceal CT with colonic contrast is then utilized as a problem solving technique for equivocal cases after this initial assessment. PMID- 12744503 TI - The CT diagnosis of acute appendicitis. AB - In clinically equivocal cases of acute appendicitis, CT performed after the opacification of the terminal ileum and the cecum offers a significant improvement in diagnostic accuracy when compared to diagnosis based on clinical signs and symptoms alone. The method employed to opacify the bowel has been the subject of discussion in the literature. We believe that the oral administration of contrast, without the use of rectal or intravenous contrast, provides the best combination of diagnostic accuracy and convenience. The CT signs of acute appendicitis are described. Ways of minimizing the difficulties of diagnosing a ruptured appendix are discussed. Because of the radiation involved, CT should not be used routinely in acute appendicitis, but should be reserved for clinically equivocal cases. PMID- 12744505 TI - Coronary artery disease: new insights into the pathophysiology, prevalence and early detection of a monster menace. AB - Coronary artery disease (CAD) is the leading cause of morbidity and mortality in the United States and industrialized countries. In the undeveloped world a similar epidemic is brewing. A new pathophysiologic paradigm has emerged, which assigns the mediators of inflammation a much larger role in the disease process. This paradigm has helped explain the unpredictable nature of many adverse consequences of CAD. The long latent phase of the disease and often sudden initial presentation make efforts at early detection extremely important. Considerable work has been devoted to identify as well as influence predisposing risk factors for developing arteriosclerosis. Novel markers of inflammation, like C-reactive protein, have been identified and compared to traditional risk factors. In addition, new imaging modalities introduce the possibility of screening for sub-clinical disease. Electron-beam and spiral CT scanners, as well as other techniques, are emerging as powerful tools to detect early disease presence and allow intervention to take place before major clinical events occur. Advances in our understanding of the pathophysiology and our ability to image the stages of silent disease will go hand in hand to revolutionize our approach to prevention and treatment of this deadly disease. PMID- 12744504 TI - Review of suspected acute appendicitis in adults and children using CT and colonic contrast material. PMID- 12744506 TI - Nitrogen comes down to earth: report from the 5th European Nitrogen Fixation Conference. AB - For four days and four nights, with almost 50 presentations and more than 175 posters, the 5th European Nitrogen Fixation Conference continued a tradition of excellence, bringing scientists from diverse fields such as microbiology, biochemistry, computational genomics, and plant physiology together to address the complex problems associated with biological nitrogen fixation (BNF). The conference was hosted by the John Innes Center and the University of East Anglia in Norwich, England and took place from September 6 through 10, 2002. A diverse range of topics was presented, from the evolution of rhizobial genomes to the plant genes involved in bacterial and fungal symbiosis, to the structure of nitrogenase, and to the means by which nitrogen is shuttled between the symbiotic bacteria and the plant. Additionally, sessions involving broader issues, such as nitrogen fertilizer use and work being done in developing countries, brought home the importance of the research being carried out in BNF around the world. PMID- 12744507 TI - A profile of putative parasitism genes expressed in the esophageal gland cells of the root-knot nematode Meloidogyne incognita. AB - Identifying parasitism genes encoding proteins secreted from a nematode's esophageal gland cells and injected through its stylet into plant tissue is the key to understanding the molecular basis of nematode parasitism of plants. Meloidogyne incognita parasitism genes were cloned by microaspirating the cytoplasm from the esophageal gland cells of different parasitic stages to provide mRNA to create a gland cell-specific cDNA library by long-distance reverse-transcriptase polymerase chain reaction. Of 2,452 cDNA clones sequenced, deduced protein sequences of 185 cDNAs had a signal peptide for secretion and, thus, could have a role in root-knot nematode parasitism of plants. High throughput in situ hybridization with cDNA clones encoding signal peptides resulted in probes of 37 unique clones specifically hybridizing to transcripts accumulating within the subventral (13 clones) or dorsal (24 clones) esophageal gland cells of M. incognita. In BLASTP analyses, 73% of the predicted proteins were novel proteins. Those with similarities to known proteins included a pectate lyase, acid phosphatase, and hypothetical proteins from other organisms. Our cell specific analysis of genes encoding secretory proteins provided, for the first time, a profile of putative parasitism genes expressed in the M. incognita esophageal gland cells throughout the parasitic cycle. PMID- 12744508 TI - Isolation of a premycorrhizal infection (pmi2) mutant of tomato, resistant to arbuscular mycorrhizal fungal colonization. AB - Arbuscular mycorrhizae (AM) represent an ancient symbiosis between mycorrhizal fungi and plant roots which co-evolved to exhibit a finely tuned, multistage interaction that assists plant growth. Direct screening efforts for Myc- plant mutants resulted in the identification of a tomato (Lycopersicon esculentum L. cv. Micro-Tom) mutant, M20, which was impaired in its ability to support the premycorrhizal infection (pmi) stages. The Myc- phenotype of the M20 mutant was a single Mendelian recessive trait, stable for nine generations, and nonallelic to a previously identified M161 pmi mutant. The M20 mutant was resistant to infection by isolated AM spores and colonized roots. Formation of Glomus intraradices appressoria on M20 roots was normal, as on wild-type (WT) plants, but in significantly reduced numbers. A significant reduction in spore germination was observed in vitro in the presence of M20 exudates relative to WT. Our results indicate that this new mutant shares similar physiological characteristics with the M161 pmi mutant, but has a more suppressive Myc- phenotype response. PMID- 12744509 TI - Peh production, flagellum synthesis, and virulence reduced in Erwinia carotovora subsp. carotovora by mutation in a homologue of cytR. AB - Erwinia carotovora subsp. carotovora is a causal agent of soft-rot diseases in a wide variety of plants. Here, we have isolated a new regulatory factor involved in the virulence of E. carotovora subsp. carotovora by in vivo insertional mutagenesis using a transposon Tn5. The gene was homologous to cytR encoding a transcriptional repressor of nucleoside uptake and catabolism genes in Escherichia coli, Salmonella typhimurium, and Vibrio cholerae. Phenotypic characterization of a nonpolar deletion mutant of the cytR homologue (delta cytR) revealed that the delta cytR mutant produced a reduced level of polygalacturonase (Peh) and lost its motility compared to that in the parental strain. With electron microscopy, the delta cytR mutant was shown to be aflagellate. Furthermore, the expression of fliA and fliC (encoding sigma28 and flagellin, respectively) was also reduced in delta cytR mutant. The virulence of delta cytR mutant was reduced in Chinese cabbage and potato compared to that of the parental strain. These results suggest that the CytR homologue of E. carotovora subsp. carotovora positively controls Peh production and flagellum synthesis and plays an important role in its pathogenicity. PMID- 12744510 TI - An investigation into the involvement of defense signaling pathways in components of the nonhost resistance of Arabidopsis thaliana to rust fungi also reveals a model system for studying rust fungal compatibility. AB - Seventeen accessions of Arabidopsis thaliana inoculated with the cowpea rust fungus Uromyces vignae exhibited a variety of expressions of nonhost resistance, although infection hypha growth typically ceased before the formation of the first haustorium, except in Ws-0. Compared with wild-type plants, there was no increased fungal growth in ndr1 or eds1 mutants defective in two of the signal cascades regulated by the major class of Arabidopsis host resistance genes. However, in the Col-0 background, infection hyphae of U. vignae and two other rust fungi were longer in sid2 mutants defective in an enzyme that synthesizes salicylic acid (SA), in npr1 mutants deficient in a regulator of the expression of SA-dependent pathogenesis related (PR) genes, and in NahG plants containing a bacterial salicylate hydroxylase. Infection hyphae of U. vignae and U. appendiculatus but not of Puccinia helianthi were also longer in jar1 mutants, which are defective in the jasmonic acid defense signaling pathway. Nevertheless, haustorium formation increased only for the Uromyces spp. and only in sid2 mutants or NahG plants. Rather than the hypersensitive cell death that usually accompanies haustorium formation in nonhost plants, Arabidopsis typically encased haustoria in calloselike material. Growing fungal colonies of both Uromyces spp., indicative of a successful biotrophic relationship between plant and fungus, formed in NahG plants, but only U. vignae formed growing colonies in the sid2 mutants and cycloheximide-treated wild-type plants. Growing colonies did not develop in NahG tobacco or tomato plants. These data suggest that nonhost resistance of Arabidopsis to rust fungi primarily involves the restriction of infection hypha growth as a result of defense gene expression. However, there is a subsequent involvement of SA but not SA-dependent PR genes in preventing the Uromyces spp. from forming the first haustorium and establishing a sufficient biotrophic relationship to support further fungal growth. The U. vignae Arabidopsis combination could allow the application of the powerful genetic capabilities of this model plant to the study of compatibility as well as nonhost resistance to rust fungi. PMID- 12744511 TI - Two potato proteins, including a novel RING finger protein (HIP1), interact with the potyviral multifunctional protein HCpro. AB - Potyviral helper-component proteinase (HCpro) is a multifunctional protein exerting its cellular functions in interaction with putative host proteins. In this study, cellular protein partners of the HCpro encoded by Potato virus A (PVA) (genus Potyvirus) were screened in a potato leaf cDNA library using a yeast two-hybrid system. Two cellular proteins were obtained that interact specifically with PVA HCpro in yeast and in the two in vitro binding assays used. Both proteins are encoded by single-copy genes in the potato genome. Analysis of the deduced amino acid sequences revealed that one (HIP1) of the two HCpro interactors is a novel RING finger protein. The sequence of the other protein (HIP2) showed no resemblance to the protein sequences available from databanks and has known biological functions. PMID- 12744512 TI - A protein kinase from Colletotrichum trifolii is induced by plant cutin and is required for appressorium formation. AB - When certain phytopathogenic fungi contact plant surfaces, specialized infection structures (appressoria) are produced that facilitate penetration of the plant external barrier; the cuticle. Recognition of this hydrophobic host surface must be sensed by the fungus, initiating the appropriate signaling pathway or pathways for pathogenic development. Using polymerase chain reaction and primers designed from mammalian protein kinase C sequences (PKC), we have isolated, cloned, and characterized a protein kinase from Colletotrichum trifolii, causal agent of alfalfa anthracnose. Though sequence analysis indicated conserved sequences in mammalian PKC genes, we were unable to induce activity of the fungal protein using known activators of PKC. Instead, we show that the C. trifolii gene, designated LIPK (lipid-induced protein kinase) is induced specifically by purified plant cutin or long-chain fatty acids which are monomeric constituents of cutin. PKC inhibitors prevented appressorium formation and, to a lesser extent, spore germination. Overexpression of LIPK resulted in multiple, abnormally shaped appressoria. Gene replacement of lipk yielded strains which were unable to develop appressoria and were unable to infect intact host plant tissue. However, these mutants were able to colonize host tissue following artificial wounding, resulting in typical anthracnose lesions. Taken together, these data indicate a central role in triggering infection structure formation for this protein kinase, which is induced specifically by components of the plant cuticle. Thus, the fungus is able to sense and use host surface chemistry to induce a protein kinase-mediated pathway that is required for pathogenic development. PMID- 12744513 TI - Flagellin from an incompatible strain of Acidovorax avenae mediates H2O2 generation accompanying hypersensitive cell death and expression of PAL, Cht-1, and PBZ1, but not of Lox in rice. AB - Acidovorax avenae causes a brown stripe disease in monocot plants. We recently reported that a rice-incompatible strain of A. avenae caused hypersensitive cell death in rice and that the flagellin of the incompatible strain was involved in this response. The incompatible strain induced the rapid generation of H2O2 accompanying hypersensitive cell death and the expression of defense genes such as PAL, Cht-1, PBZ1, and LOX, whereas the compatible strain did not. The purified incompatible flagellin also induced the expression of PAL, Cht-1, and PBZ1, but LOX expression was not induced by the incompatible flagellin. PAL and LOX enzymatic activities were increased by inoculation with the incompatible strain, whereas only PAL activity was increased by the incompatible flagellin. Interestingly, the flagellin-deficient incompatible strain lost the ability to generate H2O2 and induce hypersensitive cell death, but PAL, Cht-1, and PBZ1 expression still were induced by inoculation with the deficient strain, suggesting that induction of these genes is regulated not only by flagellin but also by some other signal. Thus, the incompatible flagellin of A. avenae is a specific elicitor in rice, but it is not the only factor capable of inducing the rice defense system. PMID- 12744514 TI - Transcriptional silencing of geminiviral promoter-driven transgenes following homologous virus infection. AB - Promoters isolated from the Tomato leaf curl virus (TLCV) drive both constitutive and tissue-specific expression in transgenic tobacco. Following systemic TLCV infection of plants stably expressing TLCV promoter:GUS transgenes, transgene expression driven by all six TLCV promoters was silenced. Silencing in the TLCV coat protein promoter:GUS plants (V2:GUSdeltaC) was characterized in more detail. Transgene silencing observed in leaf, stem, and pre-anthesis floral tissue occurred with the continued replication of TLCV in host tissues. Infection of the V2:GUSdeltaC plants with heterologous geminiviruses did not result in transgene silencing, indicating that silencing was specifically associated with TLCV infection. Nuclear run-on assays indicated that silencing was due to the abolition of transcription from the V2:GUSdeltaC transgene. Bisulfite sequencing showed that silencing was associated with cytosine hypermethylation of the TLCV derived promoter sequences of the V2:GUSdeltaC transgene. Progeny derived from V2:GUSdeltaC plants silenced by TLCV infection were analyzed. Transgene expression was silenced in progeny seedlings but was partially reactivated in the majority of plants by 75 days postgermination. Progeny seedlings treated with the nonmethylatable cytosine analog 5-azacytidine or the histone deacetylase inhibitor sodium butyrate exhibited partial reactivation of expression. This is the first report of the hypermethylation of a virus-derived transgene associated with a DNA virus infection. PMID- 12744515 TI - A chorismate mutase from the soybean cyst nematode Heterodera glycines shows polymorphisms that correlate with virulence. AB - Parasitism genes from phytoparasitic nematodes are thought to be essential for nematode invasion of the host plant, to help the nematode establish feeding sites, and to aid nematodes in the suppression of host plant defenses. One gene that may play several roles in nematode parasitism is chorismate mutase (CM). This secreted enzyme is produced in the nematode's esophageal glands and appears to function within the plant cell to manipulate the plant's shikimate pathway, which controls plant cell growth, development, structure, and pathogen defense. Using degenerate polymerase chain reaction primers, we amplified and cloned a chorismate mutase (Hg-cm-1) from Heterodera glycines, the soybean cyst nematode (SCN), and showed it had CM activity. RNA in situ hybridization of Hg-cm-1 cDNA to SCN sections confirms that it is specifically expressed in the nematodes' esophageal glands. DNA gel blots of genomic DNA isolated from SCN inbred lines that have differing virulence on SCN resistant soybean show Hg-cm-1 is a member of a polymorphic gene family. Some Hg-cm family members predominate in SCN inbred lines that are virulent on certain SCN resistant soybean cultivars. The same polymorphisms and correlation with virulence are seen in the Hg-cm-1 expressed in the SCN second-stage juveniles. Based on the enzymatic activity of Hg-cm-1 and the observation that different forms of the mutase are expressed in virulent nematodes, we hypothesize that the Hg-cm-1 is a virulence gene, some forms of which allow SCN to parasitize certain resistant soybean plants. PMID- 12744516 TI - Ectopic expression of Ralstonia solanacearum effector protein PopA early in invasion results in loss of virulence. AB - Ralstonia solanacearum OE1-1 (OE1-1) is pathogenic to tobacco. The type III secreted effector protein popA of OE1-1 showed 97.6% identity to popA of R. solanacearum GMI1000, which is not pathogenic to tobacco. Reverse transcription polymerase chain reaction analysis showed that popA in OE1-1 was expressed at 3 h after inoculation (HAI), but not before, in infiltrated-tobacco leaves. Pathogenicity analysis using a popABC operon-deleted mutant of OE1-1 (deltaABC) showed that popABC is not directly involved in the pathogenicity of OE1-1. When Papa, which constitutively expresses popA, was infiltrated into tobacco leaves, popA was expressed by 0.5 HAI. Papa could no longer multiply or spread in tobacco leaves and was no longer virulent. Moreover, the hypersensitive response (HR) and expression of HR-related genes were not induced in Papa-infiltrated leaves. Papa was also avirulent in a tobacco root-dipping inoculation assay. These results suggest that the expression of popA in Papa immediately after invasion triggers the suppression of bacterial proliferation and movement, resulting in loss of virulence. However, Papa retained its virulence when directly inoculated into xylem vessels. This result suggests that tobacco plants can recognize PopA when it is expressed early in disease development, and respond with an effective defense in the intercellular spaces. PMID- 12744517 TI - Nonspecific lipid-transfer protein genes expression in grape (Vitis sp.) cells in response to fungal elicitor treatments. AB - Nonspecific lipid transfer proteins (nsLTPs) are small, basic cystein-rich proteins believed to be involved in plant defense mechanisms. Three cDNAs coding nsLTPs from grape (Vitis vinifera sp.) were cloned by reverse-transcriptase polymerase chain reaction (RT-PCR) and PCR. The expression of nsLTP genes was investigated in 41B-rootstock grape cell suspension, in response to various defense-related signal molecules. Ergosterol (a fungi-specific sterol) and a proteinaceous elicitor purified from Botrytis cinerea strongly and rapidly induced the accumulation of nsLTP mRNAs. Jasmonic acid, cholesterol, and sitosterol also promoted nsLTPs mRNA accumulation, although to a lesser extent, whereas salicylic acid had no effect. High performance liquid chromatography analysis indicated that the amounts of three LTP isoforms (previously named P1, P2, and P4) were increased by ergosterol. None of the four isoforms displayed any significant antifungal properties, with the exception of the P4 isoform, which reduced Botrytis mycelium growth in vitro, but only in calcium-free medium. The results are discussed in the context of plant-pathogen interactions. PMID- 12744518 TI - Ultrastructure and development of Pleistophora ronneafiei n. sp., a microsporidium (Protista) in the skeletal muscle of an immune-compromised individual. AB - This report provides a detailed ultrastructural study of the life cycle, including proliferative and sporogonic developmental stages, of the first Pleistophora species (microsporidium) obtained from an immune-incompetent patient. In 1985, the organism obtained from a muscle biopsy was initially identified as belonging to the genus Pleistophora, based on spore morphology and its location in a sporophorous vesicle. Since that initial report, at least two new microsporidial genera, Trachipleistophora and Brachiola, have been reported to infect the muscle tissue of immunologically compromised patients. Because Trachipleistophora development is similar to Pleistophora, and as Pleistophora was only known to occur in cold-blooded hosts, the question of the proper classification of this microsporidium arose. The information acquired in this study makes it possible to compare Pleistophora sp. (Ledford et al. 1985) to the known human infections and properly determine its correct taxonomic position. Our ultrastructural data have revealed the formation of multinucleate sporogonial plasmodia, a developmental characteristic of the genus Pleistophora and not Trachipleistophora. A comparison with other species of the genus supports the establishment of a new species. This parasite is given the name Pleistophora ronneafiei n. sp. PMID- 12744519 TI - A model of biocomplexity and its application to the analysis of some terrestrial and marsh eukaryotic microbial communities with an emphasis on amoeboid protists. AB - Biocomplexity theory has become increasingly important in understanding ecosystem dynamics as we realize that the interactions among subunits in a multi-component system often produce elaborate states that are not easily explained in terms of the individual parts of the system. A Euclidean geometric model of biocomplexity is presented and illustrated using protistan communities. The model is based on three quantitative biotic dimensions (indices) for small subsamples (0.01 g) taken from each sample core of substratum: (1) richness of morphospecies expressed as mean count per 0.01 g, (2) spatial diversity of protists expressed as then umber of unique morphospecies (i.e. those occurring in only one of the 0.01-g subsamples and not in any of the other subsamples), and (3) patchiness (non-uniform aggregation) of the distribution of protists across the 0.01-g subsamples. These three indices are mapped into a three-dimensional Euclidean space model, and the position of each point and its geometric distance from the origin are used as a general index of biocomplexity. The usefulness of the model is illustrated by applying it to a range of terrestrial and marsh communities. Within the set of 15 samples examined in this study, the marsh rhizosphere samples are among the most complex. PMID- 12744520 TI - Presence of glyoxylate cycle enzymes in the mitochondria of Euglena gracilis. AB - Isocitrate lyase and malate synthase are specific enzymes of the glyoxylate cycle, used here as glyoxysomal markers. Both enzymes were found in the mitochondrial fraction after organelle fractionation by isopycnic centrifugation. Electron microscopy of this fraction indicated that mitochondria were the only recognizable organelles. Using an immunogold labeling method with anti-(malate synthase) antiserum, the only organelles stained in cells were the mitochondria. These results show that the glyoxylate cycle is present in mitochondria in Euglena. PMID- 12744522 TI - Ciliated protozoa in the rumen of Turkish domestic cattle (Bos taurus L.). AB - Rumen contents obtained from 28 domesticated cattle (Bos taurus L.) slaughtered at abatoirs near Izmir, Turkey were surveyed for ciliate protozoa. Protozoa are known to make an appreciable contribution to ruminal fiber digestion in many different geographical areas; however, little if any information is available on their occurrence in Turkish cattle. As a result of our survey, 13 genera including 52 species were identified. Nine of the species were further divided into 36 forma. The average ciliate density in our cattle (52.44 X 10(4)/ml) was higher than that of Turkish domestic sheep and other domestic cattle reported previously from different geographical areas. Entodinium basoglui, Entodinium williamsi f. williamsi, E. williamsi f. turcicum, E. dalli f. rudidorsospinatum, Entodinium imai, Entodinium oektemae, Eudiplodinium dehorityi, Epidinium graini, Ophryoscolex purkynjei f. bifidobicinctus, and Ophryoscolex purkynjei f. bifidoquadricinctus have previously been reported from Turkey and appear to be endemic. All ofthe remaining species represent a new host record for domestic Turkish cattle. This study also reports for the second time the presence of Entodinium constrictum in herbivorous mammals, and is the first record of its occurrence in domesticated cattle. PMID- 12744521 TI - Heparan sulfate proteoglycans mediate the invasion of cardiomyocytes by Trypanosoma cruzi. AB - Cytoadherence is an important step for the invasion of a mammalian host cell by Trypanosoma cruzi. Cell surface macromolecules are implicated in the T. cruzi cardiomyocyte recognition process. Therefore, we investigated the role of cell surface proteoglycans during this invasion process and analyzed their expression after the parasite infected the target cells. Treatment of trypomastigote forms of T. cruzi with soluble heparan sulfate resulted in a significant inhibition in successful invasion, while chondroitin sulfate had no effect. Removal of sulfated glycoconjugates from the cardiomyocyte surface using glycosaminoglycan (GAG) lyases demonstrated the specific binding of the parasites to heparan sulfate proteoglycans. Infection levels were reduced by 42% whenthe host cells were previously treated with heparitinase II. No changes were detected in the expression of GAGs infected cardiomyocytes even after 96 h of infection. Our data demonstrate that heparan sulfate proteoglycans, but not chondroitin sulfate, mediate both attachment and invasion of cardiomyocytes by T. cruzi. PMID- 12744523 TI - An enzyme-linked immunosorbent assay (ELISA) for the identification of Naegleria fowleri in environmental water samples. AB - Naegleria fowleri, a free-living amoeba, is the causative agent of primary amoebic meningoencephalitis, a fatal human disease of the central nervous system often contracted after swimming in fresh water. Identifying sites contaminated by N. fowleri is important in order to prevent the disease. An Enzyme-Linked ImmunoSorbent Assay (ELISA) has been developed for the specific identification of N. fawleri in primary cultures of environmental water samples. Of 939 samples isolated from artificially heated river water and screened by ELISA, 283 were positive. These results were subsequently confirmed by isoelectric focusing, the established reference method. A sensitivity of 97.4% and a specificity of 97% were obtained. These results indicate that this ELISA method is reliable and can be considered as a powerful tool for the detection of N. fowleri in environmental water samples. PMID- 12744524 TI - Dexiotrichides pangi n. sp. (protozoa, ciliophora, scuticociliatia), a new marine ciliate from the north China sea. AB - The morphology, infraciliature, and silverline system of a new marine scuticociliate, Dexiotrichides pangi n. sp. were investigated. The new species is characterized by: size about 45-65 x 20-25 microm in vivo with kidney-like body shape and obliquely truncated semicircle-shaped apical plate; cytostome at bottom of conspicuously depressed oral cavity, which is located at the cell equatorial level; paroral membrane extending anteriorly to membranelle 3; scutica multi rowed; 33-38 somatic kineties; contractile vacuole near ventral side and subcaudally positioned, opening at posterior end of somatic kinety 3; one oval macronucleus and one small micronucleus; caudal cilium positioned in a small pouch; marine habitat. Based on the data obtained, an improved diagnosis for the genus Dexiotrichides is suggested: body with circular cross-section and conspicuous cilia-free apical plate; buccal cavity conspicuously depressed with cytostome located near or at equatorial level; three membranelles transversely orientated each with 2-3 rows; paroral membrane zigzaging structure, extending to about half of the length of buccal field; multi-rowed scutica; somatic kinety one strongly shortened and terminating anteriorly at posterior end of buccal field; basal bodies in equatorial region arranged usually in circular pattern, while in the anterior portion of somatic kinety 2, basal bodies characteristically in pairs and separated from the posterior part of kinety 2; one caudal cilium. PMID- 12744525 TI - Permanent expression of a cyclin B homologue in the cell cycle of the dinoflagellate Karenia brevis. AB - The eukaryotic cell cycle is driven by a set of cyclin-dependent kinases associated with their regulatory partners, the cyclins, which confer activity, substrate specificities and proper localization of the kinase activity. We describe the cell cycle of Karenia brevis and provide evidence for the presence of a cyclin B homologue in this dinoflagellate using two antibodies with different specificities. This cyclin B homologue has an unusual behavior, since its expression is permanent and it has a cytoplasmic location throughout the cell cycle. There is no evidence for translocation to the nucleus during mitosis. However, it appears also to be specifically bound to the nucleolus throughout the cell cycle. The permanent expression and the cytoplasmic localization during mitosis of this cyclin B homologue is similar to p56, a cyclin B homologue previously described in a different species of dinoflagellate, Crypthecodinium cohnii. Here we discuss this unusual behavior of the cyclin B homologue in dinoflagellates, its relationship to the unusual characteristics of dinomitosis, and its potential implications regarding the evolution of cell cycle regulation among eukaryotes. PMID- 12744526 TI - Screening of substrate analogs as potential enzyme inhibitors for the arginine kinase of Trypanosoma cruzi. AB - Arginine kinase catalyzes the transphosphorylation between phosphoarginine and ADP. Phosphoarginine is involved in temporal ATP buffering and inorganic phosphate regulation. Trypanosoma cruzi arginine kinase phosphorylates only L arginine (specific activity 398.9 x mUE-min(-1) x mg(-1)), and is inhibited by the arginine analogs, agmatine, canavanine, nitroarginine, and homoarginine. Canavanine and homoarginine also produce a significant inhibition of the epimastigote culture growth (79.7% and 55.8%, respectively). Inhibition constants were calculated for canavanine and homoarginine (7.55 and 6.02 mM, respectively). In addition, two novel guanidino kinase activities were detected in the epimastigote soluble extract. The development of the arginine kinase inhibitors of T. cruzi could be an important feature because the phosphagens biosynthetic pathway in trypanosomatids is different from the one in their mammalian hosts. PMID- 12744527 TI - Freshwater foraminiferans revealed by analysis of environmental DNA samples. AB - Sediment-dwelling protists are among the most abundant meiobenthic organisms, ubiquitous in all types of aquatic ecosystems. Yet, because their isolation and identification are difficult, their diversity remains largely unknown. In the present work, we applied molecular methods to examine the diversity of freshwater Foraminifera, a group of granuloreticulosan protists largely neglected until now. By using specific PCR primers, we detected the presence of Foraminifera in all sediment samples examined. Phylogenetic analysis of amplified SSU rDNA sequences revealed two distinct groups of freshwater foraminiferans. All obtained sequences branched within monothalamous (single-chambered), marine Foraminifera, suggesting a repeated colonization of freshwater environments. The results of our study challenge the traditional view of Foraminifera as essentially marine organisms, and provide a conceptual framework for charting the molecular diversity of freshwater granuloreticulosan protists. PMID- 12744528 TI - Molecular cloning and sequencing of the merozoite surface antigen 2 gene from Plasmodium falciparum strain FCC-1/HN and expression of the gene in mycobacteria. AB - Strain bacillus Calmette-Guerin (BCG) of Mycobacterium bovis has been used as a live bacterial vaccine to immunize more than 3 billion people against tuberculosis. In an attempt to use this vaccine strain as a vehicle for protective antigens, the gene encoding merozoite surface antigen 2 (MSA2) was amplified from strain FCC-1/HN Plasmodium falciparum genome, sequenced, and expressed in M. bovis BCG under the control of an expression cassette carrying the promoter of heat shock protein 70 (HSP70) from Mycobacterium tuberculosis. The recombinant shuttle plasmid pBCG/MSA2 was introduced into mycobacteria by electroporation, and the recombinant mycobacteria harboring pBCG/MSA2 could be induced by heating to express MSA2; the molecular mass of recombinant MSA2 was about 31 kDa. This first report of expression of the full-length P. falciparum MSA2 gene in BCG provides evidence for use of the HSP70 promoter in expressing a foreign gene in BCG and in development of BCG as a multivalent vectoral vaccine for malaria. PMID- 12744529 TI - The "price" of information. PMID- 12744530 TI - Maternal and perinatal factors influencing the duration of exclusive breastfeeding during the first 6 months of life. AB - The aim of this study was to evaluate the influence of certain factors on the duration of exclusive breastfeeding during the first 6 months of life. In 597 mothers, 26 variables were assessed during the postpartum period. The mothers were interviewed monthly by telephone about how they were feeding their babies. In 539 mothers (92.2%), complete data were obtained until the sixth month. At discharge, 1 month, 4 months, and 6 months, the frequency of exclusive breastfeeding was 97%, 83%, 56%, and 19%, respectively. The median duration was 4 months. A longer duration of exclusive breastfeeding was significantly associated with positive maternal attitudes toward breastfeeding, adequate family support, good mother-infant bonding, appropriate suckling technique, and no nipple problems. These associations persisted after controlling for maternal education and other confounding variables. Certain maternal factors related to a longer duration of exclusive breastfeeding can be identified in the maternity ward and might contribute to the development of more effective breastfeeding policies. PMID- 12744531 TI - A description of the relationship between breastfeeding experiences, breastfeeding satisfaction, and weaning in the first 3 months after birth. AB - This longitudinal study describes the relationships among breastfeeding experiences, maternal breastfeeding satisfaction (measured by the Maternal Breastfeeding Evaluation Scale [MBFES]), and weaning in the first 3 months postpartum. Postal surveys were used to collect data antenatally and at 2 weeks, 6 weeks, and 3 months postpartum from 365 women recruited from 3 public hospitals in Australia. Breastfeeding problems were associated with a reduction in MBFES subscale scores (P < .05) but not with weaning. Infant breastfeeding satisfaction subscale score was negatively associated with perceived inadequate milk supply (P < .05) and positively associated with engorgement (P < .05). Significant predictors of weaning at all points were total MBFES score in the lowest tertile (adjusted odds ratios [ORs]: 2 weeks, 21.9; > 2-6 weeks, 14.6: > 6 weeks to 3 months, 6.4) and leaking milk (adjusted ORs: < or = 2 weeks, 0.23; > 2-6 weeks, 0.19; > 6 weeks to 3 months, 0.15). Perceived inadequate milk was a significantpredictoronly for weaning > 2 to 6 weeks (adjusted OR = 4.2). PMID- 12744532 TI - Long-term duration of breastfeeding in Swedish low birth weight infants. AB - The aim of this study was to describe the long-term incidence of breastfeeding in a geographically defined cohort of low birth weight (LBW) (< 2500 g) Swedish infants (N = 70) at discharge from a neonatal unit and at the postnatal ages of 2, 4, 6, and 8 months. The infants' breastfeeding data were examined retrospectively and compared to annual breastfeeding data for all infants born in the same year and county (N = 2,751). Ninety-three percent of the LBW infants were fed breast milk at discharge and 36% at 6 months, compared to 97% and 75%, respectively, of the county population. Cox regression analysis showed that multiparity was a significant predictor associated with shorter breastfeeding in LBW infants (odds ratio = 2.51. 95% confidence interval, 1.35-4.69). LBW infants had a high breastfeeding incidence but a significantly shorter duration than controls. This result indicates the need for intensified support throughout the breastfeeding period. PMID- 12744534 TI - Duration of breastfeeding in Swedish primiparous and multiparous women. AB - The aim of this study was to describe the effects of sociodemographic factors and maternity ward practices on the duration of breastfeeding in Swedish primiparas (n = 194) and multiparas (n = 294), consecutively selected from hospital birth files for 3 months, who responded to a questionnaire 9 to 12 months after childbirth. The impact of sociodemographic data and maternity ward practices on exclusive and any breastfeeding were examined. Smoking and supplementation without medical reasons influenced the duration of both exclusive and any breastfeeding negatively, whereas early first breastfeeding influenced the duration of both exclusive and any breastfeeding positively, and parity had no significant influence. Late hospital discharge influenced the duration of exclusive breastfeeding positively, and higher maternal age influenced the duration of any breastfeeding positively. These variables altogether explained 11.4% (P < .001) of the variance in the duration of exclusive breastfeeding and 8.2% (P < .001) of the duration of any breastfeeding. PMID- 12744533 TI - The baby-friendly hospital initiative increases breastfeeding rates in a US neonatal intensive care unit. AB - This study evaluated the impact of a Baby-Friendly designation on breastfeeding rates in a US neonatal intensive care unit (NICU). The medical records of all surviving infants directly admitted to the Boston Medical Center's level III, 15 bed NICU in 1995 (before Baby-Friendly policies were implemented) and 1999 (when Baby-Friendly status was granted) were reviewed. Infants receiving any breast milk by any means during the first week of enteral feeds were considered to have initiated breastfeeding. Maternal and infant demographics for 1995 and 1999 were comparable. The NICU breastfeeding initiation rate increased from 34.6% (1995) to 74.4% (1999) (P < .001). Among 2-week-old infants, the proportion receiving any breast milk rose from 27.9% (1995) to 65.9% (1999) (P < .001), and the proportion receiving breast milk exclusively rose from 9.3% (1995) to 39% (1999) (P = .002). The implementation of Baby-Friendly policies leading to a Baby-Friendly designation was associated with increased breastfeeding initiation and duration rates. PMID- 12744536 TI - Is it possible for a breastfed baby to be overweight? PMID- 12744535 TI - Response of breasts to different stimulation patterns of an electric breast pump. AB - To test the effect on milk ejection, an electric breast pump was programmed to provide pumping patterns with frequencies of 45 to 125 cycles/min and vacuums of 45 to -273 mm Hg. The time taken for milk ejection to occur (measured using ultrasound to detect a dilation of a lactiferous duct in the opposite breast) in response to the current Medela electric breast pump pattern (45 cycles/min) was 147 +/- 13 s. For patterns that more closely resemble the sucking frequency of an infant when it first attaches to the breast, milk ejection occurred between 136 +/- 12 and 104 +/- 10 s, although this difference was not statistically significant. Milk ejection in response to breastfeeding occurred after 56 +/- 4 s. The applied vacuum affected the amount of milk that was removed up to 50 to 70 s after milk ejection but not the time for milk ejection. PMID- 12744537 TI - Promoting the exclusive feeding of own mother's milk through the use of hindmilk and increased maternal milk volume for hospitalized, low birth weight infants (< 1800 grams) in Nigeria: a feasibility study. AB - A feasibility study was used to determine (1) if hindmilk feedings of own mother's milk, as reported in the United States, could be instituted in a Nigerian neonatal intensive care unit and result in adequate infant weight gain without exogenous additives; and (2) if the use of a hospital-grade electric breast pump to separate foremilk from hindmilk was feasible in this setting. Mean weight gain for 16 preterm infants during the hindmilk intervention (18.8 g/d) exceeded intrauterine standards without the use of exogenous substances. At the time of hospital discharge, mean maternal daily milk volume was 342 mL/kg/d, indicating that the infants, on average, had 90% more milk available to them than they required. Mean infant weight gains were 14.2 and 16.6 g/d from 1 to 7 and 8 to 23 (x = 14.8) days postdischarge, respectively. Therefore, hindmilk feedings are effective and feasible for hospitalized, low birth weight infants in developing countries, and lactation specialists may use our protocol for further research. PMID- 12744538 TI - Breast milk donors in France: a portrait of the typical donor and the utility of milk banking in the French breastfeeding context. AB - Although information regarding attitudes and characteristics of human blood donors has been researched, little is known about the motivations and demographic and personality characteristics of women who choose to donate their breast milk. Eight milk banks in France participated in a study examining donor characteristics, providing data on 103 women. The results showed that the donors were women of average childbearing age with strong support at home. Almost half did not work outside of the home, compared to the national average of 80% of women in this age group; similarly, a large number (currently working or not) were from the health and social services fields. Reasons for donation were largely altruistic, and a general optimistic attitude prevailed within the participants. The results of this study provide useful information for the recruitment of potential donors as well as information on how to facilitate and provide optimal service through milk donation. PMID- 12744539 TI - Rounding alone: assessing the value of grand rounds in contemporary departments of medicine. PMID- 12744540 TI - Strategies for improving attendance at medical grand rounds at an academic medical center. AB - OBJECTIVE: To evaluate, in this before-and-after study, the results of 5 strategies for improving attendance at medical grand rounds at a tertiary care academic medical center. METHODS: The strategies included (1) using electronic card readers to improve understanding of attendance patterns, (2) conducting yearly needs assessment surveys, (3) developing sessions of topical interest, (4) increasing formal participation by residents and faculty researchers, and (5) enhancing publicity. Attendance at medical grand rounds by Mayo Clinic faculty, fellows, residents, and others was measured by card readers between 1998 and 2001. RESULTS: After implementation of the 5 strategies, the mean +/- SD attendance (as measured by card readers) at medical grand rounds increased 39% from 99.0 +/- 24.6 persons in 1998 to 137.4 +/- 25.2 persons in 2001 (P < .001). CONCLUSION: An organized effort can improve attendance at medical grand rounds at an academic medical center. PMID- 12744541 TI - Factors influencing clinical decisions to initiate thyroxine therapy for patients with mildly increased serum thyrotropin (5.1-10.0 mIU/L). AB - OBJECTIVE: To determine factors that influence clinicians' decisions to treat subclinical hypothyroidism. PATIENTS AND METHODS: We studied patients living within 120 miles of our institution in Rochester, Minn, whose serum thyrotropin value was between 5.1 and 10.0 mIU/L from January 1, 1995, through December 31,1996. Exclusion criteria were history of thyroid disease or prior abnormal thyrotropin level and fewer than 5 follow-up visits or fewer than 2 follow-up thyrotropin measurements. Of the 2655 patients in this cohort, only 55% (1466) had measurement of thyroid microsomal antibodies (TMAb). From this group, records of 539 patients were selected randomly according to antibody status (238 with positive, 199 with negative, and 102 with no TMAb measurement). After exclusion of patients receiving thyroxine replacement and other medications affecting thyroid function, patients with postpartum thyroiditis, and patients in the intensive care unit, data from 450 patients were analyzed. RESULTS: Thyroxine therapy was prescribed for 39% of the patients with thyrotropin levels between 5.1 and 10.0 mIU/L. In multivariate analysis, TMAb status (P < .001), index thyrotropin (P < .002), and free thyroxine (FT4) (P = .03) were the strongest predictors for the decision to treat. Fifty-nine percent of the patients with positive TMAb were treated compared with 24% of those with negative TMAb and 30% of those who did not have a TMAb test. The mean thyrotropin level in the treated group was 7.1 mIU/L compared with 63 mIU/L in the untreated group. The mean FT4 level was 1.08 ng/dL in the treated group who had this measured compared with 1.16 ng/dL in the untreated group. Treated patients were younger than untreated patients (56.5 years vs 60.6 years), and the proportion of female patients was higher in the treated group compared with the untreated group (75.8% vs 65.2%); however, neither of these differences was significant after correction for the multiple comparisons. In multivariate analysis neither age nor sex was a significant predictor of treatment, but in the subgroup comparison, patients aged 31 to 50 years were twice as likely to be treated compared with those aged 61 to 80 years. CONCLUSION: Among patients with serum thyrotropin levels of 5.1 to 10.0 mIU/L, those with positive TMAb values, those with higher thyrotropin values, and those with lower FT4 values were more likely to receive thyroxine therapy. Younger patients were more likely to be treated than older patients. PMID- 12744542 TI - Effect of diabetes on the mortality risk of cardiogenic shock in a community based population. AB - OBJECTIVE: To examine the mortality of diabetic vs nondiabetic patients with anterior myocardial infarction (AMI) among the subsets of this population who did and did not develop cardiogenic shock. PATIENTS AND METHODS: The study population consisted of a consecutive series of 1263 Olmsted County, Minnesota, patients admitted to the coronary care unit at the Mayo Clinic in Rochester, Minn, between January 1, 1988, and July 31, 2000. Of these patients, 73 met the criteria for cardiogenic shock during their hospitalization. In-hospital and postadmission mortality were compared between diabetic and nondiabetic patients within the cardiogenic shock and nonshock patient groups, respectively. RESULTS: In patients with AMI and cardiogenic shock, diabetes was associated with a trend for increased in-hospital mortality (odds ratio, 2.82; 95% confidence interval [CI], 0.90-9.92; P = .08). In 73 patients with cardiogenic shock, estimated survival at 1, 3, and 5 years was 25%, 17%, and 17%, respectively, for diabetic patients, and 50%, 44%, and 36%, respectively, for nondiabetic patients (P = .046). The association between diabetic patients and increased long-term mortality was stronger in patients with cardiogenic shock than in patients without cardiogenic shock (adjusted relative risk, 2.08; 95% CI, 1.11-3.90; P = .02). In diabetic patients without cardiogenic shock, estimated survival at 1, 3, and 5 years was low, at 75%, 61%, and 45%, respectively, compared with 83%, 76%, and 69%, respectively, for nondiabetic patients (adjusted relative risk, 1.29; 95% CI, 1.02-1.62; P = .03). CONCLUSION: The presence of diabetes as a comorbidity in patients with AMI appears to be associated with increased mortality compared with nondiabetic patients, and this relationship may be potentially magnified in patients who develop cardiogenic shock. PMID- 12744543 TI - Granular cell tumor of the sellar and suprasellar region: clinicopathologic study of 11 cases and literature review. AB - OBJECTIVE: To report a series of cases of surgically resected granular cell tumors in an attempt to better characterize their clinical presentation, imaging features, and treatment outcomes with attention to previously published literature. PATIENTS AND METHODS: This is a retrospective study of 11 patients with granular cell tumors. Information obtained from patients' medical records was supplemented by direct physician and patient telephone contact. RESULTS: The study group consisted of 9 women and 2 men, with a mean +/- SD age of 50 +/- 15 years (range, 26-73 years). Seven patients were symptomatic, 3 of whom presented with visual complaints. Four patients presented primarily with endocrine dysfunction (2) or headaches (2). The duration of symptoms varied from 1 to 12 months (mean +/- SD, 6.0 +/- 4.5 months). Formal visual field testing revealed bitemporal hemianopsia in 6 and a right-sided visual field deficit in 2. The tumor appeared as a well-defined, relatively homogeneously (6) or heterogeneously (1) enhancing suprasellar mass on contrast infusion. Tumor size varied from 1.5 to 6.0 cm (mean +/- SD, 3.1 +/- 1.6 cm). At surgery, the mass was noted to be firm and vascular, and in at least 3 patients these features prevented gross total resection. Only 1 patient who had undergone biopsy of his lesion received adjuvant radiation therapy. Follow-up was obtained in all but 1 patient. Nine of these 10 patients are alive and are either free of disease or have had no disease progression. Follow-up in these 9 patients varied from 1 to 16 years (mean +/- SD, 6.5 +/- 53 years). CONCLUSIONS: A granular cell tumor is generally a surprise finding in that it is seldom considered in the differential diagnosis of contrast enhancing, demarcated sellar region masses. Granular cell tumors are benign and slow growing with no pronounced tendency for invasion or recurrence. Because of the indolent growth of granular cell tumors, the surgical goal of tumor decompression is a reasonable therapeutic approach. PMID- 12744544 TI - Effect of breathing supplemental oxygen on motion sickness in healthy adults. AB - OBJECTIVE: To compare the effects of breathing supplemental oxygen vs air on alleviating motion sickness in healthy adults. SUBJECTS AND METHODS: Between April and July 2002, 20 healthy subjects were exposed to a provocative motion on 2 occasions (1-week interval) according to a randomized, counterbalanced, crossover design. During motion, subjects rated their nausea (1, no symptoms, to 4, moderate nausea) every 30 seconds. Once mild nausea occurred, subjects began breathing supplemental oxygen or air through a face mask. Motion ceased when moderate nausea occurred, but subjects continued breathing study gases for 5 minutes while recovering. Recovery was assessed for 20 minutes after motion. RESULTS: There were no significant differences in the rate of increase in symptom severity or the rate of recovery between the 2 conditions. CONCLUSION: Breathing supplemental oxygen had no advantage over breathing air in reducing motion sickness in healthy adults. PMID- 12744545 TI - Primer on medical genomics. Part VII: The evolving concept of the gene. AB - The draft sequence of the human genome was reported 2 years ago, and the task of filling gaps and polishing the sequence is nearing completion. However, despite this remarkable achievement, there is still no definitive assessment of the number of genes contained in the human genome. In part, this uncertainty reflects our growing understanding of the complexity and diversity of gene structure. Examples of complex gene structure are considered in the context of a discussion about the evolution of our understanding of gene structure and function. PMID- 12744546 TI - Walter Gilbert--1980 Nobel Prize for Chemistry. PMID- 12744547 TI - Hepatic veno-occlusive disease (sinusoidal obstruction syndrome) after hematopoietic stem cell transplantation. AB - Hepatic veno-occlusive disease (VOD), increasingly referred to as sinusoidal obstruction syndrome, is a well-recognized complication of hematopoietic stem cell transplantation and contributes to considerable morbidity and mortality. In the Western Hemisphere, VOD, classified as a conditioning-related toxicity, is most commonly caused by stem cell transplantation. VOD has been described after all types of stem cell transplantation, irrespective of the stem cell source, type of conditioning therapy, or underlying disease. Recognition of this disease in the posttransplantation setting remains a challenge in the absence of specific diagnostic features because many other more common conditions can mimic it. Limited therapeutic or preventive strategies are currently available for the management of VOD. In this review, we provide a comprehensive account of the pathophysiology of this disease as we understand it today, risk factors for its development, and the current state of knowledge regarding preventive and therapeutic options. PMID- 12744548 TI - Skin blood flow in adult human thermoregulation: how it works, when it does not, and why. AB - The thermoregulatory control of human skin blood flow is vital to the maintenance of normal body temperatures during challenges to thermal homeostasis. Sympathetic neural control of skin blood flow includes the noradrenergic vasoconstrictor system and a sympathetic active vasodilator system, the latter of which is responsible for 80% to 90% of the substantial cutaneous vasodilation that occurs with whole body heat stress. With body heating, the magnitude of skin vasodilation is striking: skin blood flow can reach 6 to 8 L/min during hyperthermia. Cutaneous sympathetic vasoconstrictor and vasodilator systems also participate in baroreflex control of blood pressure; this is particularly important during heat stress, when such a large percentage of cardiac output is directed to the skin. Local thermal control of cutaneous blood vessels also contributes importantly--local warming of the skin can cause maximal vasodilation in healthy humans and includes roles for both local sensory nerves and nitric oxide. Local cooling of the skin can decrease skin blood flow to minimal levels. During menopause, changes in reproductive hormone levels substantially alter thermoregulatory control of skin blood flow. This altered control might contribute to the occurrence of hot flashes. In type 2 diabetes mellitus, the ability of skin blood vessels to dilate is impaired. This impaired vasodilation likely contributes to the increased risk of heat illness in this patient population during exposure to elevated ambient temperatures. Raynaud phenomenon and erythromelalgia represent cutaneous microvascular disorders whose pathophysiology appears to relate to disorders of local and/or reflex thermoregulatory control of the skin circulation. PMID- 12744549 TI - Microscopic colitis. AB - Microscopic colitis has 2 main subtypes, collagenous colitis and lymphocytic colitis, that are similar clinically and histologically and are distinguished mainly by the presence or absence of a thickened subepithelial collagen band. Microscopic colitis accounts for approximately 10% of chronic watery diarrhea and may be associated with abdominal pain and mild weight loss. It is typically diagnosed in the sixth to eighth decade of life, and there is a female pre dominance that is more striking for collagenous colitis than for lymphocytic colitis. Endoscopic and radiographic findings of the colon appear normal. Diagnosis is made by colonic biopsy, which shows an intraepithelial lymphocytosis and a mixed inflammatory infiltrate in the lamina propria. Many potential treatments have been reported, but few have been subjected to controlled treatment trials. A stepwise approach to therapy often leads to satisfactory control of symptoms. PMID- 12744550 TI - Clopidogrel-induced systemic inflammatory response syndrome. AB - Clopidogrel bisulfate, a widely used inhibitor of platelet aggregation, is considered at least as safe as aspirin. We describe a patient who developed a systemic inflammatory response syndrome consisting of high fever, tachycardia, cellulitis-like rash, impaired liver function, and mild leukopenia after receiving clopidogrel before coronary angiography and stent implantation. The reaction resolved promptly after withdrawal of the drug and recurred shortly after a rechallenge dose was administered, thus making the diagnosis of a clopidogrel-induced reaction highly probable. Recognition of this clopidogrel induced syndrome is extremely important, both for rapid discontinuation of the offending drug and for avoidance of unnecessary drug therapy or invasive procedures. PMID- 12744551 TI - Osteoarticular infection complicating enterococcal endocarditis. AB - Despite the common occurrence of musculoskeletal complaints in patients with infective endocarditis, infectious osteoarticular complications are diagnosed infrequently. Moreover, although enterococcal infection is the third most common cause of infective endocarditis, infectious osteoarticular complications are rare. We report a case of disk space infection in a patient with enterococcal endocarditis. Blood cultures and an L3-4 aspirate grew Enterococcus faecalis, and transthoracic echocardiography revealed a large vegetation on the posterior mitral valve leaflet. The osteoarticular infection resolved with antimicrobial treatment, but worsening heart failure necessitated valve replacement surgery. The patient had an uneventful recovery with no evidence of recurrence or complications. A review of the medical literature from 1966 through 1998 identified 13 additional cases, only 8 of which provided clinical and treatment data. We present the clinical and laboratory findings reported in these cases, along with data from our patient. This report highlights the rare occurrence of osteoarticular infection in the setting of enterococcal endocarditis and emphasizes early recognition and treatment. PMID- 12744552 TI - Upper lobe pulmonary fibrosis associated with high-dose chemotherapy containing BCNU for bone marrow transplantation. AB - Upper lobe fibrotic lung disease is most often associated with sarcoidosis, Langerhans cell histiocytosis, silicosis, and other pneumoconioses but is usually not associated with drug-induced lung disease. Carmustine (BCNU) is a chemotherapeutic agent known to cause pulmonary toxicity. The radiographic pattern is usually diffuse bilateral lung disease predominantly in the lung bases. Upper lobe fibrotic disease associated with BCNU has been reported to occur in children treated for central nervous system gliomas. Often the lung disease occurs years after the exposure. Despite the widespread use of BCNU in the treatment of malignancy in adults, to our knowledge, the complication of upper lobe fibrotic disease has not been reported in adults. We describe a patient who presented with pneumothorax and bilateral upper lobe pulmonary fibrosis that we believe was due to BCNU given for bone marrow transplantation as part of therapy for breast cancer. Bilateral upper lobe pulmonary fibrosis can be associated with chemotherapeutic drugs. PMID- 12744553 TI - 78-year-old woman with fever, weight loss, and rash. PMID- 12744554 TI - Management of the social consequences of seizures. AB - People with seizures experience restrictions in many areas of life, including participation in recreational activities, social situations, employment, and driving. Health care providers and patients should be informed of the potential hazards faced by people with seizures. The patient and practitioner who know the potential risks can make rational decisions about participation in these activities. Risk assessment and participation should be individualized according to the patient's unique situation. Understanding the inherent risk of a particular activity, risk of recurrence of seizures, legal restrictions (some activities may be governed by law or specific code), seizure type, and adverse effects of medication may be useful in making decisions concerning participation in a wide variety of activities. The information in this article should be used to maximize the patient's quality of life. PMID- 12744555 TI - Multiple blood donations and iron deficiency in patients with restless legs syndrome. PMID- 12744556 TI - Long-term survival of patients with anorexia nervosa. PMID- 12744557 TI - Serial measurement of serum tryptase in angiotensin-converting enzyme inhibitor associated angioedema. PMID- 12744558 TI - Cataract surgery output and cost of hospitalization for cataract surgery in the University of Benin Teaching Hospital. AB - Cataract is the most common cause of curable blindness in Nigeria. Nigeria has an overall population of approximately 110 million with a blindness prevalence rate of 1%. Cataract is responsible for 30 - 60% of the blindness. A great deal of blindness prevention activity should therefore centre around cataract surgery if we are to clear out cataract back log of over 1/2 a million individuals. Only 106 cataract surgeries were performed in the 24 months reviewed. There were 75 males and 31 females. This study highlights the meagre contribution of teaching hospitals to the prevention of blindness. Several factors including ignorance, poverty, socioeconomic and political tensions and teaching hospital bureaucracy are no doubt responsible for this. Hospitalization for cataract surgery is becoming very unpopular in the developed world as this tends to increase cost of surgery. The need to establish cataract surgery outreach services and adopt day case surgical procedures in our hospitals cannot be over emphasized as strategies for clearing our national cataract backlog. PMID- 12744559 TI - Correlates of emergency response interval and mortality from severe anaemia in childhood. AB - A retrospective study to determine the influence of blood transfusion emergency response interval on Mortality from childhood severe anaemia was carried out. Admission records of all children with severe anaemia over a 5-year period was reviewed. Those who either died before transfusion or got discharged against medical advice were excluded. A total of 5790 patients were admitted during the 5 year period. Ten percent (10%) had severe anaemia. Malaria, the leading cause of anaemia in this series, was found in 80% of the patients. Twenty patients (3.3%) died before transfusion while 50 discharged from the hospital against medical advice. There was male preponderance. Ages 2-5 years were the peak age group for severe anaemia. No patient had haematocrit less than 5% but 20 (4.2%) had heaematocrit above 20%. The hospital stay for majority (74.8%) of the patients was 72 hours or less. Mortality (Case fatality) increases with increase in transfusion emergency response interval within 24 hours. Based on the transfusion emergency response interval versus mortality curve, a mortality risk assessment scores were derived for use in clinical practice to determine the risk of dying from the disease. We recommend that national or hospital policy on blood transfusion be enunciated to ensure that all patient with severe anaemia get transfused within 2 hours of diagnosis. PMID- 12744560 TI - A comparison of the efficacy of alfentanil and remifentanil analgesic infusions for spinal surgery. AB - The use of alfentanil infusion was compared with that of remifentanil infusion for spinal cord surgery in a retrospective review. The aim was to compare the outcome when methohexitone was used as the only hypnotic agent in the two groups. Over a 3-year period, 5 patients (group 1) had Alfentanil infusion and 11 patients (group 2) had remifentanil infusion for analgesia during spinal cord surgery. Results showed that remifentanil lead to a faster onset of recorvery than alfentanil. It also provided better haemodynamic stability than alfentanil without excesive hypotension (p > 0.05). Our experience here indicated that remifentanil provided better flexibility of use with less tachycardia and respiratory depression than alfentanil for spinal surgery. PMID- 12744561 TI - HIV-1 infection in adults with haematological malignancies in Yaounde, Cameroon. AB - To determine the association between haematological malignancies and the HIV-1 in Yaounde, Cameroon, adult patients (> 16 years) followed up in the Haematology Clinics of two major hospitals were screened for the HIV between 1994 and 1999. There were nine haematological malignancies diagnosed among the 172 patients including Non Hodgkin's lymphomas (31.9%); chronic lymhocytic leukaemia (21.5%); chronic myelogeneous leukaemia (18.0%); acute myelogeneous leukaemia (9.9%); acute lymphoblastic leukaemia (7.6%) and multiple myeloma (7.0%). Burkitt's lymphoma, Hodgkin's disease and myelodysplastic syndrome were less frequently diagnosed. Forty-five of all cases (26.2%) had antibodies to the HIV-1 virus, predominantly in patients with Non-Hodgkin's lymphomas (p < 0,001, OR = 5.8, adjusted for age; CI = 2.7 - 12.4). About 19.4% and 11.8% of cases with chronic and acute myelogenous leukaemia respectively were HIV-1 positive. Although B lineage-derived malignancies are more often associated with the HIV infection, other malignant proliferations of the haematopoietic system may not be coincidental. PMID- 12744563 TI - Leprosy control in the post leprosaria abolition years in Nigeria: reasons for default and irregular attendance at treatment centres. AB - A questionnaire was administered to all patients with leprosy seen at the four leprosy clinics in Anambra State in a face to face interview. The questions covered, among other items, the clinic attendance behaviour and the single most important reason, monthly, for absenteeism in the preceding year. The total and individual frequencies of the reasons for absenteeism were determined for the various behavioural subgroups. The differences in frequencies and associations were analysed. Values of P < 0.05 were considered as significant. The results showed that 27 females and 26 males were interviewed. 39.6% of the patients were irregular attenders 735% were defaulters. Attendance at meetings (P < .001); work at home (P < 0.01) fear/shame/indignation (P < 0.05); no confidence in treatment (P < 0.025) were significant reasons for absenteeism among irregular attenders inter-current illnesses as reasons for absenteeism did not differ significantly between regular and irregular attendees. The association between clinic attendance behaviour and lesion location (revealed Vs concealed) was not statistically significant (X(2)0.3). The findings in this study indicate that in the post leprosaria abolition years, default and irregular clinic attendance by patients with leprosy are numerically large and may compound the problems of control programmes, and thus negate the realization of the global goal of intercepting leprosy transmission. PMID- 12744562 TI - Sources of sexual information and its relevance to sexual behaviour in Nigeria. AB - A study was carried out to identify the various sources of sexual information by adolescents in Nigeria and their influence on the sexual behaviours of the subjects, using: a) coitus prior to marriage b) expectation with first coitus c) freeness to discuss with spouse or anybody as parameters. The study was carried between 1997 and 1998 using subjects randomly selected from three Nigerian communities, viz: Enugu, Benin and Nnewi. Coitus before marriage was significantly higher in those who got their first ever information from peers than those who got it from other sources (P < 0.01). Fulfillment of expectation with first coitus was also significantly higher among those who were taught by parents, peers and teachers than those who sought their information on their own from books, magazines and films (p < 0.05). However, in considering their ability to discuss with anybody, this was found to be significantly higher in those who sought information on their own than those who got their first information by personal contact with parents, peers and teachers (p < 0.05). The latter was found to be more inhibited from discussing sexuality with their spouse or anybody than those who got their information from books/magazines and films. Sex education of adolescents should, therefore, be provided in a cultural, community based setting of which the guardian programme should be only one component. It may be counter-productive in Nigeria if the adolescents continue to learn about sexuality on their own from books, magazines and films. PMID- 12744564 TI - The use of simple diffusion tube samplers for the measurement of nitrogen dioxide in an operating room using nitrogen oxide as an anaesthetic (July - November 1999). AB - Estimates have been made of the amounts of nitrogen dioxide (NO2) in the operating room of University of Benin Teaching Hospital (UBTH) where nitrous oxide (N2O) a potential source of NO is used as an anaesthetic agent Measurements were made using palmes diffusion tubes, a device which is capable of taking samples of NO2 gas from the atmosphere through diffusion or permeation of this compound to the interior and subsequently trapping it by means of adsorption on reactive material, but which does not involve the active movement of the gas through the sampler. Results obtained indicate a low concentration of NO2 in the operating room with a minimum of 5.83 microg/m3 and a maximum concentration of 6.22 microg/m3 NO2. This result therefore suggests that the use of nitrous oxide in the operating room does not contribute significantly to the concentration of NO2. PMID- 12744565 TI - Schwannoma of the left brachial plexus mimicking a cervicomediastinal goiter in a young Nigerian lady. AB - The schwannoma is thought to arise from the schwann cells of the nerve sheath. This tumor is usually solitary and may arise from any cranial or peripheral nerve. It is encapsulated and appears to arise focally on a nerve trunk so that the nerve itself is stretched over the tumor rather than running through it as in neurofibroma. This report is unusual as the tumor started as a cervical swelling which subsequently grew into the mediastinum simulating a retrosternal goiter. The patient, a 25 year-old female was referred to the University College Hospital, Ibadan, 24 hours after an attempted thyroidectomy at a private hospital. The history was of a painless anterior neck swelling of 4 years duration devoid of symptoms of hyperthyroidism with associated dysphagia and weakness of the left hand. Examination showed an asthenic young woman. Her voice was hoarse but there were no eye signs suggestive of thyrotoxicosis. On the anterior neck was a sutured skin-crease scar over a diffuse anterior neck swelling which one could not get below. The left hand showed wasting of the thenar and hypothenar eminences. Thyroid function test results were within normal limits, indirect laryngoscopy showed a left vocal cord paralysis, packed cell volume was 38%. Her chest x-ray showed a huge left retrosternal and apical soft tissue mass displacing the trachea to the right (figure 1). A fine needle aspiration cytology was reported as a chronic lymphocytic thyroiditis. A presumptive diagnosis of thyroid carcinoma with retrosternal extension was made. At surgery, manipulation of the mass was difficult as the tissue was soft, slimy and ruptured easily. Severe hemorrhage was encountered necessitating a median sternotomy to control the bleeding vessels. Her post-operative period was stormy, however she thereafter made gradual progress to warrant her discharge six weeks post surgery. PMID- 12744566 TI - Effects of oral contraceptives on total serum proteins, albumin, globulins and cholesterol levels in Ibadan, Nigeria. AB - Total serum protein, albumin, globulin, albumin/globulin ratio and cholesterol levels were determined in 25 subjects on oral contraceptives and 25 controls. The mean serum total protein, globulin and cholesterol levels were significantly increased in oral contraceptive and their control counterparts. The albumin/globulin ratio in subjects on oral contraceptives users is significantly decreased compared with controls. In view of the findings of this study, it is suggested that the biochemical profile of long-term oral contraceptive users be assessed periodically. PMID- 12744567 TI - The prevalence of glaucoma in an onchoendemic community in South-Eastern Nigeria. AB - METHODOLOGY: With the aim of examining all the adults aged 30 years and above a survey was conducted in Alum-Inyi, a mesoendemic Community in South-Eastern Nigeria. The people were subjected to basic ophthalmic tests for the diagnosis of glaucoma namely corrected and uncorrected visual acuity at 6 metres using an illiterate E-chart, with pinhole assessment if visual acuity < 6/18; direct ophthalmoscopy to assess the vertical cup-disc ratio through undilated pupils and Schiotz indentation tonometry. In addition, static visual field analysis using the MK 1 Friedman visual field analyser was carried out in any glaucoma suspect. RESULTS: Of 664 eligible persons examined (out of the estimated total of 946), 14 definite cases of glaucoma were identified, giving a prevalence of 2.10% in the 30 years of age and older population. The mean vertical cup-disc ratio for the non-glaucoma cases were 0.30 (+0.004) and 0.31 (+/- 0.004) and 0.31 (+/- 0.09) in the right and left eyes respective. Among the glaucoma cases, the average vertical cup-disc ratio in the right and left eye were 0.63 (+/- 0.05) and 0.70 (+/- 0.06 respectively. No physiological cup of > or = 0.6 was recorded in our study. The mean iOP in the right and left eyes respectively were 18.54 (+/- 0.15) and 19.42 (+/- 0.14) in the nonglaucomatous cases and 26.39 (+/- 1.75) and 27.46 (+/- 2.59) in the glaucoma cases. CONCLUSION: Despite its limitations, this study confirms that the prevalence of glaucoma varies from one African population or population of African origin to another. It agrees with other reports that the risk of glaucoma increases with age. PMID- 12744568 TI - The prevalence of hand pain in Ibadan--implications for the carpal tunnel syndrome. AB - Hand pain or brachialgia paraesthetica nocturna (BPN) is a series of symptoms described as "waking up at night due to unpleasant sensations in the fingers" (Dekrom et al, 1992). Usually, these symptoms form the first rungs up the ladder leading up to the carpal tunnel syndrome (CTS) (Skandalakis et al, 1992), and thus are used as part of the diagnostic criteria for the disorder. We describe the results of a questionnaire survey of 422 patients attending two major hospitals in Ibadan, Nigeria for complaints unrelated to hand pathology. Questions related to the bio-data (including type of occupation), general health status, past medical and/or surgical history, and finally hand symptoms of the patients. For a patient to be positive for BPN, the hand symptoms had to correspond to the cutaneous distribution of the median nerve. Positive cases formed 12.75% of all interviewees in one hospital and 19.59% of respondents in the other, with females forming the bulk of those affected. This figure is somewhat higher than the ones previously reported. From earlier work on the predictive value of BPN for the carpal tunnel syndrome we conservatively estimate that between 20% and 38% of individuals in our study group will suffer from CTS. The relationship of heavy hand-use to BPN was a negative one as most sufferers of BPN were light hand-users. However, individual occupations requiring highly repetitive flexion-extension wrist movements recorded many BPN cases. No BPN case was confirmed when the patients presented in hospital, indicating that more awareness of the symptoms and signs of this disorder needs to be created among doctors and the general public. Also, we believe that more investigations, particularly among the general population, are needed to be able to define the prevalence of this condition more accurately. PMID- 12744569 TI - Prevalence of cardiovascular risk factors in an African, urban inner city community. AB - With the epidemiological transition phenomenon, more countries are expected to move from a disease pattern dominated by infectious diseases to one characterised by non-communicable diseases. Many developing countries are contending with infectious diseases as well as non-communicable diseases, yet little is known about the prevalence of cardiovascular risk factors in poor urban communities in developing countries. The objective of this community based study was to determine the prevalence of selected cardiovascular risk factors in an urban inner city community which had been followed up prospectively from 1993 to 1998. Results show that the prevalence of hypertension (Blood Pressure BP > 160/95 mm Hg) was 12.4 percent with an age-adjusted rate of 7.4 percent. This is higher than what is found in the rural parts of the country but much lower than what is generally observed in industrialized countries of the world. Though there was no significant difference in the proportion with hypertension by gender (P > 0.05), the mean systolic BP was significantly higher for men (123.9 +/- 23.9) mm Hg) than for women (120.6 +/- 26.8 mg Hg) t = 2.93, p < 0.01. The mean diastolic BP was similarly higher for men (75.8 +/- 14.9) mm Hg than women (74.0 +/- 14.9) mm Hg t = 2.76 p < 0.01). Eight percent of the study population were obese (BMI > 30), and generally, more women than men were obese. However, at ages 15-19 significantly more males than females were obese (X2 = 73, p < 0.01). Both mean systolic and diastolic BP increased with increasing weight. When adjusting for gender, the association between hypertension and other CVD risk factors remained similar for males and females and gender was not a modifier of the factors. Of the CVD risk factors studied, Age > 40 years and obesity, were significantly associated with the presence of hypertension in this community (P < 0.0001 and P < 0.0001), smoking, alcohol intake and gender were not (P > 0.05). It is necessary to implement a national plan for the control of cardiovascular diseases in order to reduce and keep to minimum cardiovascular diseases and its complications in the country. PMID- 12744570 TI - Caesarean section and birth weight at Korle Bu Teaching Hospital--preliminary report. AB - OBJECTIVE: This preliminary retrospective survey was done to find out whether the indications for Caesarean Section had any bearing on the birth weight. METHOD: The foetal outcome for 673 parturients who delivered at the Korle Bu Teaching Hospital between September 1, 1998 and December 31, 1998 was analysed. RESULT: Caesarean section done for cephalopelvic disproportion and for 2 or more previous sections yielded the highest mean birth weight (3.43 kg +/- 0.02 kg) and the best foetal outcome. Hypertensive disorders yielded the lowest mean birth weight (1.8 kg +/- 0.3 kg) and poorest foetal outcome. The mean parity and age of the parturients were similarly distributed. CONCLUSION: The birth weights appeared to vary with the indications for Caesarean section. Confounding factors such as the gestational age, parity and age of the parturients need to be controlled in a prospective study in future. Good antenatal supervision could improve on the birth weights. PMID- 12744571 TI - Serum hepatitis C virus and hepatitis B surface antigenaemia in Nigerian patients with acute icteric hepatitis. AB - Acute hepatitis is common in Nigeria and hepatitis B virus (HBV) infection has been a major aetiological factor. However, the role of Hepatitis C Virus (HCV) infection is yet undetermined. Forty-five consecutive Nigerian patients with Acute Icteric hepatitis (AIH) attending the Medical Clinic of the University College Hospital, Ibadan, Nigeria and 45 healthy adult Nigerians (controls) were studied for evidence of infection with both viruses. Questionnaire on risk procedures which predispose to acquisition of both HBV and HCV infections were administered to the patients. Blood samples were collected from all the subjects and tested for antibody to HCV (Anti-HCV) and Hepatitis B surface Antigen (HBsAg) using the second generation Enzyme Linked Immunoassay (Monolisa -R, Sansofi, Pasteur; France). Anti-HCV was detected in 21(47%) and 17(38%) of the patients and controls respectively. The corresponding prevalences of HBsAg were 38(84%) and 11(24%), p < 0.001. Hepatitis B virus infection was found to occur more than HCV infection in the patients with AIH but similar among the controls. Combined HBV and HCV infection occurred more frequently among the patients (42.1%) than in the control (11%) (; < 0.001). Although there was no significant difference in the HCV infection between the two groups, isolated HCV infection is commoner in the control than in the patients with AIH, (p < 0.001). Similarly, single HCV infection is commoner than lone HBV infection among the control, p < 0.05. In summary, this study shows that while both HBV and HCV infections are common in Nigeria, AIH may be more associated with HBV than HCV in the country. PMID- 12744573 TI - The pattern of stab injuries in Port Harcourt. AB - All cases of stabbing reporting to the casualty Department of the University of Port Harcourt Teaching Hospital (UPTH) between 1st January 1997 and 31st December, 1997 were studied prospectively. Fifty new cases reported within the study period. Thirteen cases required admission while 37 cases were treated and discharged from the casualty department. The majority of injuries were minor and most victims were male and of these, students in the age group 21 - 30 years formed the greatest number. The most common weapon used was broken bottle and injuries involving multiple anatomic regions were in the majority. All cases requiring major surgical intervention reported within 6 hours of injury. No mortality was recorded. PMID- 12744572 TI - Incidence and pattern of congenital dislocation of the hip in Aseer region of Saudi Arabia. AB - OBJECTIVE: The aim of this paper is to determine the incidence, pattern, predisposing risk factors, treatment modalities and outcome of congenital dislocation of the hip (CDH) in the Aseer region of Saudi Arabia. METHODS: A retrospective study of 300 cases of CDH seen during a 4-year period between 1996 to 1999 was carried out at the Aseer Central Hospital, Abha. The relevant data examined included the details on pregnancy, birth, family history, methods of diagnosis and treatment. Data on live births for the same period was also obtained from the Directorate of Health, Aseer Region, Ministry of Health, Saudi Arabia. RESULTS: During the period of study, 300 children were found to have CDH with an incidence rate of 3.5/1000 live births. Only 32.4% of CDH was diagnosed in first 6 months of life. Mean age at diagnosis was 14.5 +/- 19.7 months while the mean age at treatment was 44.50 +/- 36.41 months. Some 235 cases (78.3%) were females (M:F ratio = 1:3.6) and 292 (97.7%) were Saudi nationals. There was a positive family history in 64 cases (21.3%). Both hip joints were involved in 151 cases (50.3%), the left hip joint 82 cases (27.3%) and the right hip joint in 67 cases (22.3%). Delivery was by spontaneous vaginal delivery in 268 cases 89.3%), caesarean section in 28 cases (9.3%) and breech delivery in 29 cases (10%). Limping and waddling gait were the most common clinical presentation seen in 166 cases (55.3%). In 22 children (7.3%), the parents were blood relatives. First born children constituted 56 out of 216 (25.9%). In the present series, 46% of the children were treated surgically, 42% were treated conservatively and 12% were treated by both. Avascular necrosis (AVN) of the femoral head following the treatment was seen in 6 children (2%). CONCLUSION: Incidence rate of CDH in Aseer Central hospital and by inference in Aseer region of Saudi Arabia was found to be 3.5/ 1000 live births. Since the neonatal screening of CDH in this region is poor, awareness programmes, routine neonatal hip joint examination at birth and up to one year of age and plain x-ray of pelvis after the age of 3 months in high risk babies are strongly recommended. PMID- 12744574 TI - Infections caused by Acinetobacter species and their susceptibility to 14 antibiotics in Lagos University Teaching Hospital, Lagos. AB - Acinetobacter spp are well recognised as causes of nosocomial infections particularly in patients with immature or defective body defence system. Information concerning these organisms are lacking in this environment. For this reason the pattern of infection and the antimicrobial susceptibility profiles of these organisms isolated over a one-year period were studied. A total of 58 (3%) of the 2001 isolates from all clinical specimens received in the laboratory during the year were Acinetobacter spp. The 58 Acinetobacter spp constituted 5.5% of all the 1051 NLF-GNB isolated, and caused 4.6% of all the 1261 nosocomial infections. Thirty-seven (63%) and 17 (30%) of the Acinetobacter isolates were from wound infections and UTI respectively. All the infections were nosocomially acquired and were associated with compromised host immunity, defective body defence, surgery or urinary catheterization; with Acinetobacter baumannii being the predominant species. There was an apparent male predominance over females by a ratio of 1. 9:1 in the infections, particularly from 45 years and above. One hundred percent and 96.6% of the isolates were susceptible to cefoperazone sulbactam and travofloxacin respectively. Forty-five (77.6%) were susceptible to cefotaxime, 49 (84.5%) to ampicillin-sulbactam, 34 (58.6%) to ceftazidime, 38 (65.6%) to ticarcillin-clavulanic acid and 41 (70.7%) to ciprofloxacin. Generally the Acinetobacter spp showed multiple resistance to the range of antibiotics tested. All the isolates produced beta-lactamase. PMID- 12744575 TI - Spinal cord injuries in Ilorin, Nigeria. AB - BACKGROUND: Spinal Cord Injuries (SCI) usually result from road traffic accidents (RTA), falls, sports and some misadventures. This study was carried out to examine the aetiology of SCI in Ilorin, Nigeria; factors contributory to morbidity and mortality and to suggest measures for reducing them. METHODS: Age, sex, mechanism of injury, complications, duration of treatment and eventual outcome of patients admitted for SCI from 1995 to 1999 were restrospectively studied. RESULTS: Thirty-nine patients, age 19 to 60 years (mean 37.3), 36 males and three females were seen. Cervical spine injuries accounted for 46.2% of the cases. Road traffic accidents caused 67% and falls 23%. Accidents involving passengers in open lorries are associated with SCI when the goods fall on passengers as seen in five of the 26 RTA's (19.2%). More falls from kola-nut (44%) than from palm tree (11%) were observed. Limb paralysis and bladder dysfunction were the commonest complications. Ten patients died, 70% of them had cervical spine injuries. Nine of the ten deaths had multiple transfers to different centres before admission. CONCLUSION: This pattern of SCI in Ilorin, Nigeria showed that RTA has surpassed falls from trees, as the most common cause of SCI in Ilorin and probably in Nigeria. Indeed, the predominant tree implicated in this study has been kola nut tree unlike the palm tree in earlier reports. Imperative measures to improve morbidity and mortality include health education on passenger and load carriage, use of manual or motorised wheel barrow as against bearing heavy load on the head, principles of moving spinal Injured patients taught every road traveller and establishment of spinal centres and training of specialised personnel. PMID- 12744576 TI - Nasopharyngeal carriage and susceptibility patterns of Streptococcus pneumoniae in Kumasi, Ghana. AB - Penicillin resistant Streptococcus pneunmoniae poses an increasing problem in paediatrics, particularly in less developed countries. Outside of South Africa, little is known about S. pneumoniae susceptibilities in Sub-Saharan Africa. The objective of this study was to determine the prevalence of pneumococcal colonization and antimicrobial susceptibility among children in urban Ghana. METHODS: Nasopharyngeal pneumococcal colonization was examined in 311 children attending a polyclinic for sick children and an immunization clinic in Kumasi, Ghana. Isolates were tested for antibiotic susceptibility to penicillin, tetracycline, erythromycin, chloramphenicol, cefuroxime, cefotaxime, ceftriaxone, and trimethoprim-sulfamethoxazole. RESULTS: Over half (51.4%) of subjects were colonized with S. pneumoniae and 17% of isolates were resistant to penicillin, all demonstrating intermediate resistance. S. pneumoniae strains were also frequently resistant to trimethoprim-sulfamethoxazole and tetracycline, less so to chloramphenicol and cefuroxime and were almost uniformly sensitive to cefotaxime, cefriaxone and erythromycin. CONCLUSIONS: Our study shows a high rate of pneumococcal nasopharyngeal colonization and a concerning level of penicillin resistance although at a less alarming rate than seen in some other countries. Multiple antimicrobial resistance was also noted especially among drugs readily available and commonly used. These data impact treatment choices in pneumococcal disease. Vaccine may play an important role in disease limitation. An effort to curtail the misuse of antibiotics, by prescription and otherwise, may prevent further increases in resistance rates. PMID- 12744577 TI - A controlled trial of modified electroconvulsive therapy in schizophrenia in a Nigerian teaching hospital. AB - The efficacy of ECT in the treatment of Schizophrenia was investigated in a double blind controlled trial. The ICD-10 criteria for Schizophrenia were fulfilled by the 20 patients who entered the trial. Consecutive individuals who satisfied the inclusion criteria were randomly allocated to a course of (bilateral) six real or simulated ECTs each as applicable. Sixteen patients completed the ECT treatment and 20 weeks follow up period. Analysis of measures of clinical change (BPRS and SANS Scores) showed that both groups of patients improved, but the improvement of patients receiving ECT was not significantly greater than that of the control group. PMID- 12744578 TI - QRS axis deviation in Nigerian women during normal pregnancy. AB - The effect of pregnancy on the heart rate, respiratory rate, QRS axis and QRS complex duration of the ECG was investigated in 41 pregnant compared to 39 non pregnant age and height matched Nigerian subjects. Results obtained show that pregnancy had no significant effect (p > 0.05) on heart rate, respiratory rite and QRS complex duration of the ECG. However, the QRS axis showed significant (p < 0.05) left ward deviation in pregnant subjects compared to non-pregnant subjects. The magnitude of the deviation apparently increases as pregnancy progresses. In addition, chi-squared analysis revealed significant association (p < 0.001) between the incidence of left axis deviation in pregnant subjects compared to non-pregnant subjects. Our findings highlight the need for caution in the interpretation of the ECG during the antenatal period. PMID- 12744580 TI - Temporalis muscle flap and forehead flap for a single stage primary repair of the orbit after exenteration. PMID- 12744579 TI - Laboratory diagnosis of Gardnerella vaginalis vaginosis. AB - An evaluation of various laboratory detection methods and characteristics of Gardnerella vaginalis was made using high vaginal swab samples of 470 out patient clinic--attending women. Gram stain for 'clue cells' showed positive results in 118 (25.1%)cases; culture, in 100 cases, (213%) and Amine Odour (213%), in 26 cases 5.5%, Majority, 71 cases, of the culture-positive results were associated with a pH value of 6 to 7. Gardnerella vaginalis grew predominantly in enriched culture media: Modified peptone-starch dextrose blood agar used for primary culture of organism, and also proteose peptone broth + cooked meat; Brain-heart infusion broth + 5% human serum; Brain heart infusion starch agar + 5% blood, and chocolate agar. Gardnerella vaginalis culture-positive samples also exhibited positive biochemical reactions with the hydrolysis of starch sensitivity to Bacitracin and 50 microg metronidazole, and haemolysis on human blood agar. Carbohydrate fermentation test was positive for all culture-positive cases, 100% for starch and maltose only, and negative for all the cases, 0% for Mannitol and glycerol. PMID- 12744581 TI - Dermatology consultation in Olabisi Onabanjo University Teaching Hospital, Sagamu. PMID- 12744582 TI - Congenital icthyosiform--a case report. AB - Congenital icthyosis is a rare group of disorders of keratinisation. A case of this condition is reported in order to highlight the clinical features and essential components of management: resuscitation, skincare, nutrition and counselling. This case is reported in order to appraise clinicians of its presence because there was delay in the diagnosis and management due to non recognition. PMID- 12744583 TI - Klebsiella-induced purpura fulminans in a Nigerian child: case report and a review of literature. AB - Purpura fulminans (PPF) is a very severe but rare acute thrombohaemorrhagic illness of infants and young children. It occurs mainly, in patients with either congenital or acquired deficiencies of proteins C and S and antithrombin III. Features of PPF include disseminated intravascular coagulopathy, symmetrical necrotic purpura and/or ecchymoses and symmetrical peripheral gangrene; digital and/ or limb(s) amputations and end-organ failure(s) may also occur. The case of a 3.5 year-old Nigerian girl, who developed PPF following Klebsiella rhinoscleromatis septicaemia is reported to illustrate the seriousness of the disease and the need for early diagnosis and management. PMID- 12744584 TI - The conjoined twins of Gusau, Nigeria. AB - A case of Thoracopagus conjoined twins complicated with rupture of the uterus is presented. The patient was admitted after a prolonged second stage of labour. Diagnosis was made at laparotomy. Problems in diagnosis are discussed and the benefits of antenatal care and confinement under skilled supervision are emphasised. PMID- 12744585 TI - Eosinophilic enteritis--a diagnostic dilemma. AB - Eosinophilic enteritis is a rare condition of unknown aetiology, although it is generally believed to be due to intestinal allergy. It may mimic peptic ulcer, subacute (or chronic) intestinal obstruction, gastroenteritis, irritable bowel syndrome, and inflammatory bowel disease. The diagnosis is often difficult to make and most cases are only diagnosed after laparotomy/ laparoscopy and biopsy. It can be successfully treated with corticosteroids. We report a case of Eosinophilic enteritis in a 27 year old woman the symptoms of which appeared within six weeks of childbirth. With repeated episodes of abdominal pain, vomiting, occasional loose stools with weight loss, she was investigated and treated for many weeks in three hospitals without success. All investigations were inconclusive. Finally laparotomy revealed inflamed segments of small bowel, a biopsy of which showed Eosinophilic enteritis. The patient was subsequently treated successfully with Prednisolone. PMID- 12744586 TI - Hydrocephalus, ventriculo-peritoneal shunt and cerebrospinal fluid ascites. AB - A small number of patients with congenital hydrocephalus, who have been treated effectively with ventriculoperitoneal shunts, develop progressive increase in abdominal girth, due to cerebrospinal fluid ascites. This abdominal distension can produce respiratory difficulties that require endotracheal intubation and ventilator support. The respiratory difficulties and the abdominal distension were eliminated when the ventriculoperitoneal shunt was converted to a ventriculoatrial shunt in each of the three cases presented in this paper for discussion. PMID- 12744587 TI - Giant frontal sinus mucocoele with intracranial extension and orbital displacement in an elderly Nigerian. AB - Mucocoele of the frontal sinus presents with initial sign of forehead swelling in about 10% of cases, and cases with huge intracranial extension have been reported to be rare. We present a case of a giant frontal sinus mucocoele with intra cranial and intra-orbital extensions exerting a mass effect on the anterior cranial fossa in a 78 year old Nigerian female with resultant forehead swelling, proptosis and total blindness of the left eye. Diagnosis was made on clinical and radiological basis, and enucleation using coronal flap approach was done. Immediate repair of the resultant frontal bone defect was effected through the use of a curved 0.5mm stainless steel wire fixed in 3 layers across the defective frontal bone over which the soft tissues of the forehead were undermined for primary closure on sound bone. This approach was considered more appropriate than a split rib graft in view of the patient's age. No evidence of recurrence was recorded during a one-year post-surgery follow-up, suggesting that mucocoele, regardless of size can be treated with conservative surgical approach provided all cystic lining and mucocoele are removed. PMID- 12744588 TI - Overview of obesity. AB - The purpose of this article is to provide a succinct overview of obesity. It will be useful to highlight some of the epidemiological issues associated with this disease. Obesity is intertwined between public health issues, increased risk of morbidity and mortality, rising health care costs and expensive treatment options, social stigma, and internal psychological challenges--clearly no easy answers as to how to treat this health dilemma. Even with all of the challenges, bariatric surgery certainly offers a viable option for patients to get their health back, and more important, their lives back. PMID- 12744589 TI - Bariatric surgery options. AB - There are multiple surgical procedures used for weight loss. The concept of achieving malabsorption through manipulation of the gastrointestinal contract is briefly discussed. Various surgical options are considered with their advantages and disadvantages, namely vertical banded gastroplasty, gastric banding, Roux-en y gastric bypass, distal gastric bypass billiopancreatic diversion, and duodental switch. PMID- 12744590 TI - Bariatric weight loss surgery: patient education, preparation, and follow-up. AB - Bariatric weight loss surgery is no small undertaking. It requires a lifelong commitment on the part of the patient. Education is the key to helping patients achieve optimal health and a better quality of life as a result of having undergone weight loss surgery. PMID- 12744591 TI - Critical care of the morbidly obese. AB - This article describes the special needs of the critically ill morbidly obese, with a focus on the care of the postoperative obesity surgery patient. The technique of surgery is described elsewhere in this journal. Details of nursing care complicated by obesity are discussed. Environment of care modifications required for routine care of the morbidly obese in critical care are detailed. Pharmacokinetic factors to be considered are reviewed. A case study is presented to coalesce concepts presented. PMID- 12744592 TI - Obesity: impediment to wound healing. AB - Intrinsic and extrinsic factors affect wound healing. High risk factors for the obese patients include infection, seromas, anastomatic leaks, and incision dehiscence. Tissue perfusion is an issue of great concern and is a key factor in most assessments. Obesity adds another dimension to the needs of the patient and presents challenges to nurses. From routine evaluations to specialized assessments with attention to bariatric equipment needs, a thorough understanding of wound healing and potential problems of obese patients, and knowledge of interventions is needed. Nonjudgmental attitudes are imperative in planning care for the obese patients. Following a review of physiological needs and nursing interventions, a case study details one woman's surgical complications. PMID- 12744593 TI - Nutritional considerations after bariatric surgery. AB - Malnutrition is a risk that is associated with all bariatric surgeries. Malnutrition is largely preventable after these surgeries if proper patient selection, thorough preoperative nutrition education, and postoperative nutritional follow-up take place along with patient compliance. Bariatric surgery is divided into 2 major categories: restrictive or malabsorptive (with or without the restrictive aspect). The more dramatic weight loss is generally associated with procedures that are malabsorptive in nature. There is an increased risk of specific nutritional deficiencies associated with these surgeries. With proper supplementation these deficiencies are largely avoidable. This article reviews the more common bariatric surgeries and the nutritional considerations associated specifically with each surgery. The article then summarizes the typical diet advancement schedule and reviews critical care nutrition in regards to total parenteral nutrition administration for the morbidly obese individual, following bariatric surgery. PMID- 12744594 TI - Psychological aspects of delivering nursing care to the bariatric patient. AB - The purpose of this article is to discuss the physical and emotional challenges in caring for the bariatric patient and the potential biases that may complicate that process. Issues regarding fat biases, physiological and psychological components of eating, as well as nursing-patient dynamics are discussed. The intention is to show how societal pressures, personal struggles with weight, and misperceptions regarding the obese combine to subtly influence our thinking, attitudes, and subsequent behaviors. Providing a better understanding of this interplay may create more empathy for both nurses and the morbidly obese and facilitate a more comfortable atmosphere of caring for both. PMID- 12744595 TI - A personal perspective on the needs of the weight loss surgery patient. AB - Weight loss surgery (WLS) patients, such as those undergoing bileopancreatic diversions and other Roux-en-Y gastric bypass procedures, present a relatively new population for the health care system. WLS creates special needs for this population that health care providers may not be fully aware of. The author, a former WLS patient and health care quality assurance professional, presents observations from his unique personal perspective. He feels that WLS patients, and post-WLS patients receiving other health care services, can be inadvertently jeopardized by the lack of awareness of their special needs. These needs are the result of the permanent postsurgical malnutrition and malabsorption syndrome occurring after the Roux-en-Y gastric bypass. These needs include nutrition, management of electrolytes during surgery, and pain management. Additionally, the morbidly obese patients may possess low self-esteem to the degree that they may not actively and appropriately participate in their own care. The author also offers personal observations concerning his belief that there is discrimination against morbidly obese patients in the health care setting. The author presents a personal case study as a tool for better understanding of the WLS patient by caregivers. PMID- 12744596 TI - Living large: a personal story. AB - This is a personal account of the experience of a bariatric patient with a typical story of frustration with weight loss options who eventually sought bariatric surgery in desperation. Her story traces the steps of her journey to an ideal bariatric center where she derived the many benefits of competent medical care and personal support. She has been married for 7 years is an active mother of 5-year-old twin boys and is expecting her third child in May 2003. PMID- 12744597 TI - Morbid obesity: the disease and comorbidities. AB - The purpose of this article is to discuss the disease of morbid obesity, its causes, and its impact upon an individual's life. Medical personnel must have a clear understanding that morbid obesity is a disease largely due to genetic makeup and not a personality defect or lack of control. The intention is to show how morbid obesity contributes to many comorbidities that are costly, decreases quality of life, and is life-threatening. Providing a better understanding will hopefully facilitate more compassionate care among nurses who care for this population. PMID- 12744598 TI - Building blocks for polyamide nucleic acids: facile synthesis using potassium fluoride doped natural phosphate as basic catalyst. AB - Potassium fluoride doped natural phosphate, inexpensive and environmentally friendly catalyst, is shown to be an efficient basic catalyst for the N1/N9 alkylation of different nucleobases as synthons for PNAs. PMID- 12744599 TI - Synthesis and biological activities of 2-functionalized purine nucleosides. AB - Novel purine nucleosides functionalized at the 2-position have been prepared using new applications of synthetic methodology. The target molecules were designed as potential inhibitors (as their monophosphates) of the enzyme, inosine monophosphate dehydrogenase (IMPDH), and representative inhibition data are presented. Antiviral data of the compounds are discussed. PMID- 12744600 TI - Synthesis of antisense oligonucleotides with minimum depurination. AB - The removal of 4,4'-dimethoxytrityl (DMTr) groups from oligonucleotides at low pH and the acid lability of the glycosidic linkage of purine nucleotides constitute an inherent conflict in preparative oligonucleotide chemistry. The use of a mildly acidic NaOAc buffer (10 mM, pH 3.0-3.2) allows adjustment of the pH in a range where the progress of the DMTr removal reaction can be monitored conveniently by HPLC and the optimum reaction time can be calculated. As a result, oligonucleotides with minimum depurination are obtained. PMID- 12744602 TI - A new method for the synthesis of 2'-O-benzyladenosine using Mitsunobu reaction. AB - A new method to introduce a benzyl group onto the 2'-OH of purine ribonucleoside is described. Thus, 6-chloropurine 3'-O-benzoylriboside and its 5'-O-trityl congener were condensed with benzyl alcohol using the Mitsunobu reaction to give the 2'-O-benzyl derivative. The yields were varied from 4.6 to 62.9% depending on the solvent used. The product was converted to adenosine, indicating that the stereochemistry at C-2' is retained. PMID- 12744601 TI - Synthesis and biological activity of 2-aminopurine methylenecyclopropane analogues of nucleosides. AB - Synthesis and biological activity of racemic 2-aminopurine methylenecyclopropane analogues of nucleosides 4, 5, 10 and 11 is described. One-pot alkylation elimination of 2-aminopurine (6) with dibromide 7 gave a mixture of four isomeric methylenecyclopropanes. The (E, Z)-N9 and (E, Z)-N7 isomers 8 and 9 were resolved by chromatography on silica gel. Deacetylation of 8 afforded the respective (Z) N9 and (E)-N9 isomers 4 and 10 which were separated by chromatography on silica gel. In a similar fashion, (E, Z)-N7 mixture 9 furnished (Z)-N7 and (E)-N7 isomers 5 and 11. The S-(+)-enantiomer 4 was obtained by desulfurization of (S) (+)-6-thiosynguanol (13) with Raney Ni. Compound 13 was obtained from (S)-(+)-2 amino-6-chloro derivative 12 and NaSH in methanol. Racemic analogues 4, 5, 10 and 11 were inactive against HCMV, HSV-1, HSV-2, EBV and VZV. Enantiomer (S)-(+)-4 inhibited replication of HSV-1 in BSC-1 cells (ELISA) with EC50 35 microM and it was non-cytotoxic in KB cells (CC50 > 100 microM). Compound (S)-(+)-4 was also moderately effective against VZV in HFF culture with EC50/CC50 (microM) 60/>460 and it was a substrate for xanthine oxidase. PMID- 12744603 TI - Striking ability of adenosine-2'(3')-deoxy-3'(2')-triphosphates and related analogues to replace ATP as phosphate donor for all four human, and the Drosophila melanogaster, deoxyribonucleoside kinases. AB - In extension of an earlier report, six non-conventional analogues of ATP, three adenosine-2'-triphosphates (3'-deoxy, 3'-deoxy-3'-fluoro- and 3'-deoxy-3' fluoroxylo-), and three adenosine-3'-triphosphates (2'-deoxy-, 2'-deoxy-2'-fluoro and 2'-deoxy-2'-fluoroara-), were compared with ATP as potential phosphate donors for human deoxycytidine kinase (dCK), cytosolic thymidine kinase (TK1), mitochondrial TK2, deoxyguanosine kinase (dGK), and the deoxyribonucleoside kinase (dNK) from Drosophila melanogaster. With one group of enzymes, comprising TK1, TK2, dNK and dCK (with dAdo as acceptor), only 3'-deoxyadenosine-2' triphosphate was an effective donor (5-60% that for ATP), and the other five analogues much less so, or inactive. With a second set, including dCK (dCyd, but not dAdo, as acceptor) and dGK (dGuo as acceptor), known to share high sequence similarity (approximately 45% sequence identity), all six analogues were good to excellent donors (13-119% that for ATP). With dCK and ATP1, products were shown to be 5'-phosphates. With dCK, donor properties of the analogues were dependent on the nature of the acceptor, as with natural 5'-triphosphate donors. With dCK (dCyd as acceptor), Km and Vmax for the two 2'(3')-deoxyadenosine-3'(2') triphosphates are similar to those for ATP. With dGK, Km values are higher than for ATP, while Vmax values are comparable. Kinetic studies further demonstrated Michaelis-Menten (non-cooperative) or cooperative kinetics, dependent on the enzyme employed and the nature of the donor. The physiological significance, if any, of the foregoing remains to be elucidated. The overall results are, on the other hand, highly relevant to studies on the modes of interaction of nucleoside kinases with donors and acceptors; and, in particular, to interpretations of the recently reported crystal structures of dGK with bound ATP, of dNK with bound dCyd, and associated modeling studies. PMID- 12744605 TI - Convenient synthesis of 8-amino-2'-deoxyadenosine. AB - We studied the behaviour of 8-azido-2'-deoxyadenosine and 8-bromo-2' deoxyadenosine in aqueous solutions of ammonia and primary and secondary amines. Unexpectedly, 8-Azido-2'-deoxyadenosine is converted to 8-amino-2'-deoxyadenosine in excellent yields. The use of this reaction for the preparation of 8 aminoadenine derivatives needed for the preparation of oligonucleotides carrying 8-aminoadenine is discussed. PMID- 12744604 TI - Hydrodynamic and spectroscopic studies of substrate binding to human recombinant deoxycytidine kinase. AB - Deoxycytidine kinase (dCK), a cytosolic enzyme with broad substrate specificity, plays a key role in the activation of therapeutic nucleoside analogues by their 5'-phosphorylation. The structure of human dCK is still not known and the current work was undertaken to determine its oligomeric and secondary structure. Biophysical studies were conducted with purified recombinant human dCK. The Mr determined by low-speed sedimentation equilibrium under nondenaturing conditions was 60,250 +/- 1,000, indicating that dCK, which has a predicted Mr of 30,500, exists in solution as a dimer. Analysis of circular dichroism spectra revealed the presence of two negative dichroic bands located at 222 and 209 nm with ellipticity values of -11,900 +/- 300 and -12,500 +/- 300 deg x cm2 x dmol(-1), respectively, indicating the presence of approximately 40% alpha-helix and 50% beta-structure. Circular Dichroism studies in the aromatic and far-ultraviolet range and UV difference spectroscopy indicated that binding of substrates to dCK reduced its alpha-helical content and perturbed tryptophan and tyrosine. Steady state fluorescence demonstrated that deoxycytidine (the phosphate acceptor) and ATP (the phosphate donor) bound to different sites on dCK and fluorescence quenching revealed bimodal binding of deoxycytidine and unimodal binding of ATP. Spectroscopic studies indicated that substrate binding induced conformational changes, with the result that dCK exhibited different affinities for various substrates. These results are consistent with a random bi-bi kinetic mechanism of phosphorylation of dCyd with either ATP or UTP. PMID- 12744606 TI - Structural polymorphism of telomeric DNA regulated by pH and divalent cation. AB - DNA oligonucleotides can form multi-stranded structures such as a duplex, triplex, and quadruplex, while the double helical structure is generally considered as the canonical structure of DNA oligonucleotides. Guanine-rich or cytosine-rich oligonucleotides, which are observed in telomere, centromere, and other biologically important sequences in vivo, can form four-stranded G quadruplex and I-motif structures in vitro. In this study, we have investigated the effects of pH and cation on the structures and their stabilities of d(G4T4G4) and d(C4A4C4). The CD spectra and thermal melting curves of DNAs at various pHs demonstrated that acidic conditions induced a stable I-motif structure of d(C4A4C4), while the pH value did not affect the G-quadruplex structure and stability of d(G4T4G4). The CD spectra of the 1:1 mixture of d(G4T4G4) and d(C4A4C4) indicated that the acidic conditions inhibit the duplex formation between d(G4T4G4) and d(C4A4C4). Isothermal titration calorimetry measurements of the duplex formation at various pHs also quantitatively indicated that the acidic conditions inhibit the duplex formation. On the other hand, the CD spectra and thermal melting curves of DNAs in the absence and presence of Ca2+ indicated that Ca2+ induces a parallel G-quadruplex structure of d(G4T4G4) and then inhibits the duplex formation. These results lead to the conclusion that both the pH and coexisting cation can induce and regulate the structural polymorphisms the oligonucleotides in which they form the G-quadruplex, I-motif, and duplex depending on the conditions. Thus, the results reported here indicate pivotal roles of pH and coexisting cations in biological processes by regulating the conformational switching between the duplex and quadruplexes structures of the guanine-rich or cytosine-rich oligonucleotides in vivo. PMID- 12744607 TI - Synthesis, conformation, and antiviral activity of 5-methoxymethyl-2' deoxycytidine analogs. AB - Analogs of 5-methoxymethyl-2'-deoxycytidine, MMdCyd (1) by substitution at N4 were synthesized to impart resistance against deamination. The anti HSV-1 activity and solution conformation of analogs were determined. N4-Butanoyl-MMdCyd (10) was a potent inhibitor of HSV-1 replication while N4-hexanoyl-MMdCyd (11), N4-propanoyl-MMdCyd (9) and N4-acetyl-MMdCyd (8) had good activity against HSV-1 replication. All other analogs were devoid of activity against HSV-1. PMID- 12744608 TI - Mucosal melanomas. AB - Cutaneous malignant melanoma is a common malignancy and has been increasing at an alarming rate in the United States. Sun exposure is a well-known risk factor related to this disease and much is understood regarding the etiology and epidemiology of cutaneous melanomas. In contrast, primary mucosal melanomas represent an extremely rare malignancy and do not have the same risk factors or behavior patterns. They occur in areas that have no sun exposure and solid predisposing risk factors have not been identified, making this disease very difficult to diagnose or screen for. It is usually diagnosed at a later stage and carries a poor prognosis. Identifying the differences between a primary lesion and a metastatic melanoma is often challenging, because of the lack of definitive criteria, both pathologically and clinically. The rich vascular and lymphatic network surrounding these lesions may be responsible for their aggressive behavior and poor prognosis. In addition, the obscure locations where mucosal melanomas occur are an obvious reason why these lesions often go unnoticed until symptoms develop. Recent literature has raised significant questions regarding recommended treatment strategies. Earlier reports advocated radical surgery as the mainstay of therapy; however, local recurrence and survival were unchanged whether radical surgery or local excision was performed, and the most recent data are favoring the conservative approach when appropriate. Unfortunately, a multitude of adjuvant therapies have been tried without any success. Adjuvant radiotherapy plays a role when combined with surgery, particularly in the head and neck region and female genitalia, but this is reserved for nodal and locoregionally advanced disease and has had no effect when used as a prophylactic method. It is difficult to make significant advances in treatment strategies because of the rarity of the disease. As an example, one in 75 persons born in the year 2000 will develop cutaneous melanoma in his lifetime, compared with four cases per ten million people diagnosed with mucosal melanoma per year in the United States. Possible new therapies based on new biologic and immunologic findings may have future promise on being able to impact this disease. Until then, this aggressive tumor continues to have a poor prognosis, and surgical resection continues to be the mainstay of primary therapy. PMID- 12744609 TI - Ocular melanoma. AB - This article reviews the current pathogenesis, molecular changes, diagnosis, and treatment of ocular melanomas. Ocular melanomas can involve the eyelid, conjunctiva, intraocular structures, and the orbit. The most common eye melanoma involves the uveal tract and is responsible for approximately 13% of melanoma deaths. Uveal melanomas account for 10% of all melanomas. PMID- 12744611 TI - Medical management of melanoma. AB - The treatment for malignant melanoma has undergone significant changes over the past few decades, with biological therapy playing an increasingly important role and replacing the traditional cytotoxic agents as the first-line therapy for this disease, both in advanced disease and adjuvant settings. Despite the developments of new modalities of therapy for melanoma, the outcome for patients with advanced disease remains poor. This article discusses the clinical studies that have shaped our current management of melanoma, both in the adjuvant setting, and in the metastatic setting. Additionally, successes and failures of clinical trials will be discussed, as they will guide the strategic development of future studies in the management of this aggressive disease. PMID- 12744612 TI - Radiation therapy for malignant melanoma. AB - Although surgery remains the primary treatment for patients with localized melanoma, available data indicate that there is a need for improved local regional control in situations where complete surgical resection may be difficult or when high-risk features are noted pathologically. Retrospective and phase II prospective studies have revealed that elective/adjuvant radiotherapy can significantly improve the local-regional control rate in these clinical settings. The impact of elective/adjuvant radiotherapy on the incidence of distant metastasis and overall survival has yet to be determined, however. Additionally, there remains a role for radiotherapy as a primary treatment alternative for elderly patients with large facial lentigo maligna melanoma. The optimal radiation fractionation schedule remains controversial. The hypofractionated regimen is well tolerated, has resulted in improved local-regional control as compared with historical surgical results, and is convenient for a group of patients in whom survival expectations are low. Significant improvements in outcome will require commensurate improvements in systemic disease control. The importance of local control to reduce local morbidity, however, should not be underestimated, and future research goals should include randomized clinical trials to further define the role of adjuvant irradiation alone or in combination with systemic therapy. PMID- 12744613 TI - Immunotherapy of malignant melanoma. AB - Several areas of immunotherapeutic research may ultimately improve the effectiveness of active specific immunotherapy for melanoma and other malignancies. Identification of the most relevant tumor antigens will continue to be a vital component of vaccine design. Optimizing delivery of these antigens by use of adjuvants, dendritic cells, or heat shock proteins will enhance the immunogenicity of vaccines. The use of DNA vaccines to deliver nucleotides that encode relevant antigens and immunologic molecules, such as costimulatory molecules, and the use of targeted therapy with immunocytokines have yielded promising results in animal studies. Finally, cutting-edge techniques such as quantitative polymerase chain reaction and gene/protein microarrays will be used to monitor the response to a vaccine and thereby guide management decisions. Although IFN-alpha 2b is the only FDA-approved adjuvant treatment for AJCC stage IIB/III melanoma, recent data failed to show a benefit in overall survival. For patients with AJCC stage IV melanoma, chemotherapy with dacarbazine is currently the standard of care, with modest response rates of 15% to 20%. The encouraging response rates and low toxicities that were reported in phase I/III trials suggest that active immunotherapy may prove to be the most effective adjuvant therapy. At present, there are no FDA-approved cancer vaccines for malignant melanoma, and the results of ongoing randomized phase III clinical trials are greatly anticipated. PMID- 12744610 TI - Malignant melanoma in non-Caucasians: experience from Hawaii. AB - Data from the Hawaii Tumor Registry suggest that the incidence of melanoma in the non-Caucasian population of Hawaii is not substantially different from that of the remainder of the United States. Our experience indicates that melanoma in this population, although unusual, is not rare. Although lesions on the palms and soles are more common. as are subungal melanomas, primary tumors on other skin sites account for the majority of patients with cutaneous melanoma in the non Caucasian population. The substantial difference in primary tumor thickness suggests the reported poorer outcomes for non-Caucasian patients with cutaneous melanoma may be explained, at least in part, by a delay in diagnosis. Given the evidence that preventive measures and educational efforts have dramatically impacted the diagnosis and outcome of melanoma patients, it is critical to recognize that similar efforts should be directed at the non-Caucasian population. PMID- 12744614 TI - The role of isolated limb perfusion for melanoma confined to the extremities. AB - Isolated limb perfusion with Melphalan is the best treatment option to control symptomatic multiple small in-transit metastases. When lesions are bulky, Isolated Limb Perfusion (ILP) with Tumor Necrosis Factor (TNF) + Melphalan is superior as in soft tissue sarcoma. TNF changes the pathophysiology, greatly enhances the uptake of Melphalan and destructs selectively the vasculature of large tumors. To date, ILP is not indicated in an adjuvant setting. PMID- 12744615 TI - Surgical clinical trials in melanoma. AB - Over the past 30 years, several important surgery clinical trials examining the issues of excision margins, ELND, and regional therapy have given rise to the current practice guidelines for the care of melanoma patients. The development of the SLN selective lymphadenectomy concept and the increasing use of molecular staging techniques have presented new challenges in this field. Clinical trials have been initiated to address these latest issues. In the next decade, these trials should help guide the integration of the new technologies into the surgical management of melanoma. PMID- 12744617 TI - Coping with melanoma--ten strategies that promote psychological adjustment. AB - The coping strategies discussed above are not right for everyone, but there is good evidence that they are generally helpful to patients who are dealing with cancer, including melanoma. The bottom line is that these strategies help patients feel better and stronger. They feel better because they are facing the illness squarely and working through its emotional impacts, and yet also keeping a perspective on it so that it does not define them or take over their lives. Through all the trials and challenges that cancer can bring, they are keeping their wits about them and able to carry on. They feel stronger because they have support, from other people and from within themselves. They have taken stock of their most cherished reasons for living, which strengthens and sustains them in their fight against cancer. And yet they also feel that their survival is not the only important objective; the quality of their lives and relationships, the values they live by, and their spirituality also deserve attention and effort. They have the peace of knowing that their death from cancer, if it comes to that, will not obliterate the meaning, value, and joy that their life has given to them and their loved ones. PMID- 12744616 TI - World Health Organization experience in the treatment of melanoma. AB - The method of randomized trials, as performed by the WHO Melanoma Program, has definitely been a legitimate clinical study design in patients with melanoma from about 1970 to the end of the last century. Three important results of these WHO trials have substantially influenced the approach of the clinician towards melanoma: (1) There is no role for elective regional lymph node dissection, (2) narrow local excision of the primary melanoma does not entail additional risks, and (3) adjuvant treatment with chemotherapy or immunotherapy after radical surgery for regional lymph node metastases has not, until now, shown any substantial benefit. A problem arises because randomized clinical studies require long periods of time for patients accrual and completion. Frequently their legitimacy is challenged due to the appearance of new parameters, both regarding staging (eg, the introduction of new technical method such as sentinel node biopsy) and prognosis. Therefore, it would be better to define different clinical study models to quickly test a hypothesis on a small group of selected patients in order to provide quick results. We believe that this is the future of clinical research in the new millennium, because it does not seem reasonable today to plan large clinical trials that need 10 years or more of accrual and follow-up without reaching definite conclusions. PMID- 12744619 TI - Future perspectives on malignant melanoma. AB - Emphasis for the treatment of melanoma should be shifted more to prevention and early diagnosis, because early melanoma may potentially be cured in most cases. Clinical trials are important to establish more effective adjuvant modalities against melanoma. Multifaceted aspects of micrometastasis, including differentiation of different clones with respect to the primary tumor, acquisition of adhesion molecules, and host interaction with the microscopic tumor, will shed new light on the biology and mechanism of early metastasis. New molecular and genetic tools may be used to dissect the mechanisms of lymphatic and hematogenous routes of metastasis. If such mechanisms can be understood, potential therapeutic maneuvers can be developed to prevent the process of micrometastasis. PMID- 12744618 TI - The Sydney Melanoma Unit--a multidisciplinary melanoma treatment center. AB - The undoubted success of the SMU as a specialist multidisciplinary melanoma treatment center has clearly been the result of many factors. Perhaps chief among these was the vision and commitment that led Dr. Milton to establish it in the first place, and the sharing of that vision and commitment by those who were associated with him and by those who joined the SMU later. Another vitally important element, however, has been the continuing sense of unity and purpose fostered by the weekly SMU clinical meetings, which are truly multidisciplinary, in which all staff are encouraged to participate, and at which the desirability of adherence to agreed, evidence-based treatment guidelines is emphasized. A further influential factor has been the SMU's strong commitment to clinical and basic research as a concomitant of high quality clinical care, with stimulation, encouragement, and advice provided at its monthly multidisciplinary research meetings, where all current and proposed clinical and laboratory studies are discussed. As a result of these activities, despite an ever-increasing number of people working within it, the SMU has been able to present to referring doctors, to patients, and to the community a unified commitment to the best possible patient care and to high quality clinical and laboratory research. These groups have responded by recognizing the SMU as the major referral center for melanoma in Australia, as evidenced by the steadily increasing number of patients referred to it for treatment each year. Melanoma is a more pressing health problem in Australia than elsewhere, because it is the third most common cancer in women (after breast cancer and colorectal cancer), and the fourth most common cancer in men (after prostate cancer, colorectal cancer, and lung cancer). Nevertheless the experiences of the SMU as a large multidisciplinary melanoma treatment center are likely to have relevance and application in other countries, where the incidence of melanoma is lower but continues to rise, and may within a few years approach rates currently recorded in Australia. PMID- 12744620 TI - Prevalence of asthma, asthma-like and allergic symptoms in the urban and rural adult population in Eastern Turkey. AB - There is no study assessing the prevalence rates for asthma, asthma-like symptoms, and allergic symptoms in Eastern Turkey. In this study, we investigated these rates in the urban and rural adult population of Elazig Region in Eastern Turkey. A questionnaire was prepared based on the standard inquiry form of the European Community Respiratory Health Survey. The survey included 3,591 consented adults who were randomly selected. Each questionnaire was interviewed face-to face by a physician. It was found that the cumulative prevalence rates of asthma and asthma-like symptoms of the rural and the urban respondent groups were 29.6% and 25.8%, respectively. The periodic prevalence rates of the same symptoms were 27.3% and 19.3%, respectively. The percentages of the patients living in rural and urban areas diagnosed as asthma by a physician were 5.5% and 3.1%, respectively. The proportions of patients with a history of at least one allergic disorder were 36.4% and 23.7% in the rural and urban groups, respectively. Asthma and asthma-like symptoms in patients with allergic rhinoconjunctivitis were significantly higher than those without (p < 0.01). The cumulative rates of a previous diagnosis of asthma among respondents with a history of asthma-like symptoms were 18.4% in the rural and 11.2% in the urban groups (p < 0.05). Among those who had had periodic asthma-like symptoms, the rural and urban rates of previous asthma diagnosis were 18.6% and 14.3%, respectively (p < 0.01). Unexpectedly, the prevalence rates for asthma, asthma-like and allergic symptoms in adults living in the rural areas of Elazig Region in Eastern Turkey were significantly higher than the rates in the urban population. PMID- 12744621 TI - Sensitization to Dermatophagoides pteronyssinus and Blomia tropicalis extracts and recombinant mite allergens in atopic Thai patients. AB - Mite surveys in Thailand indicated that Dermatophagoides pteronyssinus (Dp) is predominant, but so far there were no data available on Blomia tropicalis (Bt), which is prevalent in the Asia Pacific region. Skin prick testing (SPT) was performed in 40 atopic children, 45 atopic adults and 17 non-atopic volunteers. Skin reactions to Dp were found in 25/40 (62.5%) and 23/45 (51.1%); skin reactions to Bt were found in 15/40 (37.5%) and 18/45 (40%) in atopic children and adults, respectively. SPT to the major sensitizing allergens Der p 1, Der p 2, Der p 5, and Blo t 5 showed positive results in 14/40 (35%), 12/40 (30%), 1/40 (2.5%) and 4/40 (10%) of atopic children, and in 12/45 (26.7%), 13/45 (28.9%), 5/45 (11.1%), 6/45 (13.3%) of atopic adults, respectively. The results indicate that Dp is one of the major sources of allergy, while Bt is a minor one and that Der p 1 and Der p 2 are important mite allergens in Chiang Mai, Thailand. PMID- 12744622 TI - Correlation between the renal C1q deposition and serum anti-C1q antibody: a potential role of anti-C1q antibody in lupus nephritis. AB - The anti-C1q antibody has been shown to be associated with lupus patients with renal involvement. We conducted a study to determine the relationship between the serum anti-C1q titer and the renal deposition of C1q. The serum anti-C1q was measured in 26 healthy controls and 47 systemic lupus erythematosus (SLE) patients who were divided into 2 groups as non-nephritis and nephritis SLE. We analyzed the relationship between the anti-C1q titers and SLE, renal C1q staining and the WHO classification for lupus nephritis. The result revealed that the serum anti-C1q was present in 50.8% of the SLE patients, that its levels in those with renal involvement were significantly higher than in the normal control group (61.540 +/- 87.720 U/ml vs 15.750 +/- 2.530 U/ml, p = 0.005). Besides, the serum anti-C1q levels were higher in the patients with lupus nephritis with C1q deposition in the kidney tissue (66.038 +/- 91.141 U/ml vs 16.652 +/- 3.097 U/ml, p < 0.01). There seems to be evidence supporting that the autoantibody anti-C1q might play a pathogenic role in lupus nephritis. PMID- 12744623 TI - Plasma endothelin-1 in infants and young children with acute bronchiolitis and viral pneumonia. AB - Respiratory syncytial virus (RSV) Infections that occur during the first three years of life have been demonstrated to be associated with the development of childhood asthma. The mechanism of virus-triggered airway inflammation Is not fully understood. Endothelin-1 is a potent bronchoconstrictor involved in many diseases including respiratory tract infections. Infants and young children diagnosed with either viral pneumonia or acute bronchiolitis, their age ranging between 2 months and 3 years, were recruited into this study. Nasopharyngeal aspirates were taken for detection of respiratory virus by antigen immunofluorescence stain, RT-PCR analysis and viral culture. Plasma endothelin-1 (ET-1) was measured by using a commercially available enzyme-linked immunosorbent assay (ELISA). Ten of the nineteen infants and children (52%) were positive for RSV infection, one co-infected with influenza A. Nine Infants (90%) were positive for RSV subtype A. There was only one infant with subtype B. One of the RSV negative individuals was positive for influenza A. In addition, we recruited 10 patients without chronic underlying or respiratory tract illness as controls. ET 1 levels were significantly increased in RSV infection compared to the controls (3.6 +/- 1.2 and 1.2 +/- 1 pg/ml, respectively (p < 0.05). In conclusion, infants and young children who are infected with RSV have an increase in circulating plasma endothelin-1. This in turn may contribute to the subsequent development of childhood asthma. PMID- 12744624 TI - Specific IgG antibody subclasses to Angiostrongylus cantonensis in patients with angiostrongyliasis. AB - Total IgG, IgG1, IgG2, IgG3, IgG4, IgA and IgM specific antibodies against Angiostrongylus cantonensis somatic antigen were determined by enzyme-linked immunosorbent assay (ELISA) in sera from proven human angiostrongyliasis (PA) cases, clinically suspected angiostrongyliasis cases with eosinophilic meningitis (EM) and healthy control (HC). The specific IgA antibody in each of the patient groups was significantly higher than those of the HC group (p < 0.05). The mean ELISA value of the specific IgM in the PA group was not significantly different from that of the HC group (p > 0.05). However, the mean specific IgM ELISA value in the EM group was significantly higher than that of the HC group (p < 0.05). The levels of the specific IgG and IgG subclasses in both patient groups were significantly higher than in the healthy control (HC) group (p < 0.001). Major differences were evident in the distribution of the IgG subclass antibodies between the patient groups. The IgG1 antibody demonstrated the highest sensitivity and specificity while the IgM and IgA responses were generally poor in both patient groups. The levels of the specific IgG antibody subclasses possibly explain immune responses to the parasite. PMID- 12744625 TI - Flexibility in the administration schedule of varicella vaccination in healthy adolescents and young adults. AB - A clinical trial to assess the immunogenicity and reactogenicity of two doses of varicella vaccine (live attenuated Oka-strain, GlaxoSmithKline Biologicals), when either given 8 or 4 weeks apart in healthy seronegative adolescents and young adults, was conducted in Khon Kaen and Bangkok, Thailand. Contrary to seroconversion rates generally reported for this age group, in our study all subjects were already seropositive after the first vaccine dose. After the first vaccine dose, geometric mean titers (GMTs) for anti-varicella antibodies were 78.4 (median 64) for the adolescent group and 136.5 (median 128) for the young adult group. Six weeks after administration of the second dose, anti-varicella GMTs reached 331.7 (median 256) and 636.9 (median 512) for the adolescent and young adult groups, respectively, with a 4.2-4.7-fold increase from pre vaccination titers. The difference in GMTs between post-dose I and dose II was statistically significant for each group. The reactogenicity after the first and second doses of vaccination was low: no varicella rash was seen, in either the shorter or longer schedule. GlaxoSmithKline Biologicals varicella vaccine (Varilix) offered a high flexibility, administration possible at either 4 or 8 weeks interval, whilst eliciting good immunogenicity and good tolerability. PMID- 12744626 TI - Specific monoclonal antibodies to Strongyloides stercoralis: a potential diagnostic reagent for strongyloidiasis. AB - In this study, specific hybridomas secreting monoclonal antibodies (MAb) to antigen of Strongyloides stercoralis filariform larvae were produced. Specific epitopes targeted by the MAb were protein in nature and located in situ in the internal content of the filariform larvae of the parasite but not in the esophagus. The MAb reacted to the homologous antigen in an indirect ELISA but did not reveal any reaction to the SDS-PAGE separated-homologous antigen in a Western blot analysis (WB) suggesting a conformational epitope specificity. The MAb were of IgG1 isotype which is the isotype known to have high affinity to this epitope so they were used in a dot-ELISA to detect the antigen of the parasite. The assay could detect the epitopes in 78 ng or more of the crude filariform larval extract but did not reveal any positive result when applied to detect antigen in stool samples of parasitologically confirmed strongyloidiasis patients. The negative antigen test results can be explained as follows. Either the MAb were filariform stage-specific and thus did not recognize the rhabditiform larval antigen mainly contained in the patient's stool or the amounts of antigen in the stool samples were too small and/or unevenly dispersed. In the latter instance, the MAb developed in this study would have a diagnostic potential if used in an immunological test design where more volume of fresh stool sample could be accommodated in the test, e.g. a sandwich plate ELISA. PMID- 12744627 TI - Production and characterization of a monoclonal antibody against recombinant glutathione S-transferase (GST) of Fasciola gigantica. AB - A monoclonal antibody (MoAb) against a recombinant glutathione S-transferase (rGST) of F. gigantica was produced in BALB/c mice. Reactivity and specificity of this monoclonal antibody was assessed by ELISA and immunoblotting. Six stable clones, namely 3A3, 3B2, 3C6, 4A6, 4B1 and 4D6 were obtained, All these MoAb reacted with rGST and native GST at a molecular weight of 28 kDa and found to be IgG1, kappa-light chain isotypes. These MoAb cross-reacted with Schistosoma mansoni and Schistosoma japonicum antigens at molecular weights of 28 and 26 kDa, respectively, but no cross-reactions were detected with antigens of Eurytrema and Paramphistomum spp. The localization of GST in metacercaria, 7-week-old juvenile and adult F. gigantica was performed by immunofluorescence technique, using MoAb as well as polyclonal antibody (PoAb) to the native protein as probes. In general, all clones of MoAb gave similar results and the pattern was quite similar to staining by PoAb. The fluorescence was intense, which implied the presence of a high concentration of GST in the parenchymal tissue in all stages of the parasite. However, the parenchymal cells were not evenly stained which implied the existence of subpopulations of this cell type with regard to GST production and storage. In addition, in adult and juvenile stages a moderate fluorescence was present in the basal layer of the tegument, while light fluorescence was observed in the caecal epithelium, cells in the ovary, testis and vitelline gland of the adult. In the metacercaria stage, in addition to parenchymal tissue, the tegument and tegumental cells were stained relatively more intense with MoAb and PoAb than in other stages. PMID- 12744628 TI - Evaluation of in-house optimized semi-nested PCR and EIA for direct detection of mycobacterial DNA in CSF. AB - A rapid and correct diagnosis of mycobacterial infections is important for effective patient treatment. Semi-nested-PCR with Fl-16 SOL, 16SOR and 16SNSR primers based on the 16S rRNA gene, under optimized conditions, can detect 499 bp amplified products from all tested mycobacteria. The assay could detect as little as 100 fg of mycobacterial DNA except for rapid growing mycobacteria, whose detection limits ranged from 1 ng to 10 pg. The specificities of the capture probes were assessed with 96 mycobacterial strains (22 species) and 33 nonmycobacterial strains (30 species). The specificities of pAll1, pTbc1 and pMar1 were 94%, 93% and 82%, respectively, and that of pAvi1, pInt1, pChe1 and pFor1 were 100%. The pTbc1 and pAvi1 were tested with DNA from 108 CSF samples, and the sensitivity and specificity of the detection method were 56% and 84% compared to culture and patient histories. The assay should be used for rapid detection and concurrent identification of slow growing mycobacteria without parallel conventional culture verification. PMID- 12744629 TI - ANA negative (Ro) lupus erythematosus with multiple major organ involvement: a case report. AB - Anti-nuclear antibody (ANA) negative systemic lupus erythematosus (SLE) occurs in about 4-13% of SLE cases. A small group of ANA negative SLE patients with positive anti-Ro antibodies usually present with typical vasculitic skin lesions which can be associated with photosensitivity, renal disease, congenital heart block or neonatal lupus. We present a case of a persistently ANA negative patient who presented with joint pain, rashes, mouth ulcer and alopecia. Clinical diagnosis of systemic lupus erythematosus was made even though ANA was negative. She was started on steroids and went into remission. Later, she developed several episodes of convulsions associated with fever and prominent vasculitic lesions. The patient was also found to have microscopic hematuria, proteinuria, anemia and thrombocytopenia. Renal biopsy showed lupus nephritis class 1B. Due to the prominent skin lesions, we performed anti-extractable nuclear antigens (ENA) antibodies test and anti-Ro turned out to be positive. The final diagnosis was ANA negative SLE (Ro lupus) with cutaneous, renal, musculoskeletal, hematological and cerebral Involvement. PMID- 12744630 TI - Cardiac disease in children in Lebanon: the AUB-MC Children's Cardiac Registry experience. AB - OBJECTIVE: To study the epidemiology of cardiac disease in children and their outcome in Lebanon, we established a Children's Cardiac Registry Center (CCRC) at the American University of Beirut-Medical Center. DESIGN/METHODS: The CCRC included prospectively all pediatric patients with congenital heart disease (CHD) and/or acquired heart disease (AHD) who were evaluated at our center, between March 1, 1997 and July 31, 2000. RESULTS: Out of the 1000 patients with cardiac anomalies enrolled in the CCRC, 917 (91.7%) had CHD and the rest had AHD. Ventricular septal defect was the most common cardiac malformation with a relative frequency of 25.3%, followed by pulmonary stenosis (14.6%), aortic anomalies (8%), ASD (8%) and tetralogy of Fallot (7.8%). Complex cardiac lesions like HLHS, TGA and AVC had lower frequencies at 0.4%, 3.7% and 3.5% respectively. The most common AHD was rheumatic heart disease (42.2%). 34.9% of the registry patients with CHD and 10.8% with AHD underwent surgical intervention. There were 4.8% and 2.4% mortality rates in the CHD and AHD groups, respectively during the 40-month study period. CONCLUSION: The prevalence of many of the cardiac malformations in the CCRC was similar to that reported in the literature. However, some of the complex cardiac lesions were less common. The outcome of the two groups of patients is comparable to the outcome of children with cardiac malformation from developed countries. The establishment of a registry at the national level is important. Appropriate identification of the cardiac disease, its epidemiology, and outcome is of utmost importance in guiding adequate care. PMID- 12744631 TI - [Subclinical carditis during an initial attack of acute rheumatic fever: contribution of colored Doppler echocardiography and therapeutic advantages]. AB - OBJECTIVES: To determine and to evaluate valvular involvement, in particular subclinical, as confirmed by colored Doppler echocardiography (CDE) during an initial attack of acute rheumatic fever (ARF). Means of diagnosis and therapeutic implications. MATERIALS AND METHODS: Over a 7-year period, from January 1994 to December 2000, 49 patients (27 females and 22 males), with a mean age of 9.2 years (range 5-14 years), who presented with a first attack of ARF, were diagnosed on the basis of clinical data (history, physical findings, specific laboratory data, EKG, and CDE) to determine the major and minor criteria of acute rheumatic fever. All patients were reinvestigated and controlled clinically and by echocardiography two weeks to three months after the first investigation. RESULTS: Reported clinical major criteria were: Arthritis, 46 cases (94%); carditis, 27 cases (55%); erythema marginatum, 3 cases (6%); subcutaneous nodules, 3 cases (6%) and chorea, 3 cases (6%). CDE abnormalities were identified in 37 cases with cardiac involvement (75% of patients), 10 of them had subclinical evidence of valvular involvement: 6 cases with mild to moderate mitral regurgitation (MR), 2 cases with moderate aortic regurgitation (AR), and 2 cases had both mild to moderate MR and moderate AR. All patients with subclinical disease and evidence of inflammatory process (7 cases) were treated by salicylates. Repeated echocardiography for control showed disappearance of valvular insufficiency in 8 patients with subclinical valvulopathy, and aggravation was observed in 2 other patients. CONCLUSION: CDE is recommended in patients with suspicion of ARF, even in normal cardiac auscultation in order to detect an acute cardiac involvement leading to an early diagnosis. The confirmation of subclinical valvular disease should be considered as major criteria for ARF. Isolated and subclinical mitral and/or aortic regurgitations, with evidence of inflammatory process, should receive corticosteroids and be followed-up regularly, clinically and non-invasively by CDE. PMID- 12744632 TI - Pediatric renal biopsy: ambulatory versus overnight hospitalization. AB - Renal biopsy may be safely performed on selected pediatric patients in an ambulatory care setting. In order to verify the safety of this new procedure, we performed 51 renal biopsies on an ambulatory care basis over a five-year period (from December 1993 to December 1998): twenty-eight biopsies on transplanted kidneys and twenty-three of native kidneys. During the same period, 56 renal biopsies were performed during a regular hospitalization of a minimum of 24 hours. We compared the two groups: Ambulatory procedure group and overnight hospitalization group. There were no significant differences between the two groups in mean age, adequacy of renal samples and complications such as post biopsy pain or hematuria. After this encouraging experience, we are proposing the ambulatory procedure to all pediatric patients in our unit. Parents will be informed that if macroscopic hematuria is noted in the immediate post-biopsy period, hospitalization might be necessary. PMID- 12744633 TI - Value of duodenal endoscopic markers of villous atrophy. AB - Several endoscopic aspects of duodenal mucosa have been described in patients with villous atrophy (VA) and celiac disease. We assessed the accuracy of three endoscopic markers in patients referred for duodenal biopsy. METHODS: From January 1992 to June 2000 all patients undergoing upper gastrointestinal (UGI) endoscopy by a single operator (n = 4102), had the second part of the duodenum examined, without any magnification technique or addition of vital dye staining, for the presence of three markers of VA, (1) reduction or loss of Kerkring's folds, (2) scalloped configuration of the duodenal folds, and (3) mosaic appearance of the mucosa. Biopsy samples were taken from those in whom the mucosa was abnormal (n = 51), and from those in whom it was normal but where biopsy was indicated (n = 230). RESULTS: None of the 230 endoscopically normal controls had VA. Seven out of 51 patients (14%) with an abnormal mucosa had a normal histology, the remaining 44 (86%) had some degree of VA confirmed at histology, partial VA in 11 patients (25%) and subtotal or total VA in 33 patients (75%). 63% of the patients with only one endoscopic marker had VA confirmed at histology whereas the coexistence of more than one marker was always associated with histological villous atrophy. The overall sensitivity, specificity, positive and negative predictive values of the endoscopic markers were respectively, 100%, 97%, 86%, and 100%. CONCLUSION: The appearance of the second part of the duodenal mucosa as assessed during routine UGI endoscopy, is a sensitive and specific indicator of the presence of VA on biopsy, and should alert the endoscopist to the possibility of a celiac disease. PMID- 12744635 TI - The etiologic role of the osteo-meatal complex in the pathogenesis of sinusitis: a retrospective study of clinical, CT and endoscopic findings in 143 patients. AB - The etiological role of the osteo-meatal complex (OMC) in the pathogenesis of sinus disease is a debatable issue. The charts of 143 patients who underwent endoscopic sinus surgery at the American University of Beirut Medical Center between 1992 and 1995 were reviewed. Clinical symptoms included facial pain, nasal obstruction, headache, and postnasal drip. CT scan and endoscopic findings were reviewed. The presence of OMC disease radiologically was correlated with all the clinical, nasal endoscopic and other radiologic findings (disease in the rest of paranasal sinuses, or presence of concha bullosa). Its etiological role in patients with sinusitis is discussed. RESULTS: No endoscopic findings could be statistically correlated with the OMC disease. Facial pain and postnasal drip were the only clinical symptoms that statistically correlated with the OMC disease (p-value = 0.009 and 0.008, respectively). The OMC disease correlated with the radiological evidence of sinusitis in any of the sinuses. PMID- 12744634 TI - [Acute toxicity in 50 patients with prostate cancer treated with conformal radiation therapy]. AB - PURPOSE: To report our experience with 3D conformal radiotherapy for prostate cancer. MATERIAL AND METHODS: We reviewed our first 50 patients diagnosed with prostate cancer. Median follow-up was 27 months (16-40 m). Median age 68 (52-74). T stage was: T1c = 12 ; T2a = 14; T2b = 10; T2c = 2; T3a = 10; T3b = 1 and T3c = 1. Gleason score (GS) 4-6 50% and GS 7-8 50%. Pretreatment PSA value of < 10 ng/ml 36%, 10-20 ng/ml 32% and > 20 ng/ml 32%. Forty patients received androgen ablation therapy 2 to 6 months before radiation. 3D conformal radiotherapy was used to allow a smaller amount of rectum and bladder to be in the high dose volume. An 18 Mv linear accelerator was used. The first 21 patients received 66 Gy, 28 patients received 70 Gy and one 74 Gy. RESULTS: The mean prostate volume was 45 cc for patients who received androgen ablation and 54 for the others (p = 0.02). The percentage of volume receiving more than 50 Gy (V50) was calculated for the rectum and bladder. The median V50 was 30% (10-55) for the rectum and 36% (5-70) for the bladder. Based on the RTOG grading (gr) for acute toxicity, there was no gr 3 gastrointestinal (GI) toxicity and only 1 gr 3 genitourinary (GU) toxicity. There were 9 gr 1 and 5 gr 2 GI toxicity, 10 gr 1 and 5 gr 2 GU toxicity. With our actual follow-up we have 2 late morbidities: gr 2 GU and one erectile failure. CONCLUSION: 3D conformal radiotherapy for prostate cancer has a good toxicity profile. Longer follow-up is needed to assess late toxicity and clinical outcome in this series. PMID- 12744636 TI - [Mesothelioma--asbestos in Lebanon: a problem to be considered]. AB - To estimate the incidence of pleural mesothelioma and its relationship with the occupational and environmental exposure to asbestos in Chekka region. Between 1991 and 2000, 22 cases of malignant mesothelioma were diagnosed at Hotel-Dieu de France Hospital. Eighteen cases were epidemiologically investigated. Fifteen among these 18 patients (83%) had a positive exposure history: exposure was occupational in 11 cases and environmental in 4 cases. The tumor was attributable to Eternit Company in 12 cases among the exposed 15 (80%). These 12 cases were secondary to occupational exposures in 8 and to environmental exposure in 4 cases. Mean latency period between exposition and diagnosis was 29 years. Fifteen patients died from the progression of their disease after a median survival of 8 months. The relationship between pleural mesothelioma and Eternit Company with the related occupational and environmental risk in Chekka region is obvious. The assessment of the incidence needs a national cancer registry. Despite the protective measures taken by the government since 1996, an increase in the incidence is suspected in the coming ten years because of the long latency period of the disease. PMID- 12744637 TI - High blood pressure: definitions, prognostic, and therapeutic implications. AB - Although it is well established that high blood pressure (BP) levels of 140/90 mmHg or higher are associated with increased cardiovascular morbidity and mortality it has not been as well appreciated that lower BP levels, namely those considered to be in the "normotensive" range, also confer an increased risk of cardiovascular disease. Recent data from the Framingham Heart Study have demonstrated that high normal levels (i.e. SBP = 130-139 and/or DBP = 85-89 mmHg) frequently progress to hypertension, are associated with structural and functional cardiovascular alterations, an atherogenic metabolic profile and/or a comorbid condition, and an increased risk of cardiovascular outcomes. Factors that predispose to progression to hypertension include higher SBP and body weight at baseline and weight gain. In low risk subjects with high normal BP, nonpharmacologic measures, especially salt restriction and weight reduction are often adequate to lower BP to < or = 130/80. In those with a high cardiovascular risk profile, BP should be reduced to < or = 120/80 with nonpharmacologic measures and pharmacotherapy. PMID- 12744638 TI - [Castelman's disease. Report of two cases with review of the literature]. AB - Castelman's disease, an unusual condition of unknown cause, consisting of a massive proliferation of lymphoid tissue. Three histologic variants (hyaline vascular, plasma-cell, and mixed) and two clinical types (localized and multicentric) of Castelman's disease have been described. Localized disease can be cured with surgery or radiotherapy, but complete remission in patients with multicentric disease have been achieved only with prednisone or chemotherapy given at the time of diagnosis. The aim of this study is to report two cases of retroperitoneal Castelman's disease with review of the literature. The first patient, of 36 years old, presented for abdominal pain with anorexia and weight loss. The abdomino-pelvic CT scan showed a 6 cm retroperitoneal mass. The biopsy of this lesion suspected a lymphoproliferative disease. At laparotomy total excision of mass was made and the final histology revealed a hyalino-vascular type of Castelman's disease. The second patient, of 26 years old, presented for left lombar pain with weight loss. The abdominopelvic CT Scan showed a 6.5 cm retroperitoneal mass. The biopsy of this lesion showed a Castelman's disease. At laparotomy total excision of mass was made and the final histology confirmed a hyalino-vascular type of Castelman's disease. PMID- 12744639 TI - Popliteal cyst in a patient with total knee arthroplasty: a case report and review of the literature. AB - We report a case of total knee arthroplasty (TKA) failure presenting initially as a large popliteal cyst without bony destruction in a 66-year-old woman. A foreign body gigantocellular inflammatory reaction against polyethylene wear particles was found on histology. The size of the cyst and the absence of bony destruction can be due to an exclusive early intramuscular granulomatous reaction. Progressive loosening of the femoral component and narrowing of the joint space heralded the TKA failure. PMID- 12744640 TI - Retropharyngeal hematoma: a complication of anticoagulant therapy. AB - Retropharyngeal hematoma is a fatal condition if clinically missed. The typical presentation of a huge posterior pharyngeal mass pushing the uvula anteriorly and obstructing the airway is described in the following case report. The clinical symptoms, physical findings, and radiological work-up are presented along with a literature review. PMID- 12744642 TI - CPICS: child and parents' interaction coding system in dyads and triads. PMID- 12744641 TI - [Vertebral actinomycosis: case report and review of the literature]. AB - In this article, the case of a 32-year-old man with a paravertebral actinomycosis is discussed. Initially, the diagnosis was not obvious but it was confirmed later with the repetitive radiologic procedures, the elimination of other etiologies (purulent, mycobacterial or mycotic infections and neoplasia) and the biopsy. Treatment with penicillin initially and then with tetracycline for a long term led to a very good outcome at a 3-year follow-up with a radiologic remission. Following the discussion of the case, a review of the literature concerning the paravertebral actinomycosis, its diagnostic clues and treatment is undertaken. PMID- 12744643 TI - Antimicrobial properties of basil and its possible application in food packaging. AB - Basil (Ocimum basilicum L.) is a popular culinary herb, and its essential oils have been used extensively for many years in food products, perfumery, and dental and oral products. Basil essential oils and their principal constituents were found to exhibit antimicrobial activity against a wide range of Gram-negative and Gram-positive bacteria, yeast, and mold. The present paper reviews primarily the topic of basil essential oils with regards to their chemical composition, their effect on microorganisms, the test methods for antimicrobial activity determination, and their possible future use in food preservation or as the active (antimicrobial), slow release, component of an active package. PMID- 12744644 TI - Preparation of a t,t conjugated linoleic acid methylester (CLA-Me) isomers mixture from synthetic CLA by methylation with BF3/methanol. AB - Mixtures of t,t conjugated linoleic acid methylester (t,t CLA-Me) isomers were prepared from synthetic CLA, consisting of 47.8% t10,c12 CLA; 45.5% c9,t11 CLA; 2.0% t,t CLA; and 4.7% others, by methylation with BF(3)/methanol (designated TT TC/CT) in conjunction with purification at -68 degrees C for 24 h. The amount or composition of the TT-TC/CT was greatly affected by the concentration of BF(3) in methanol and the duration of methylation. The methylation of 50 mg of synthetic CLA for 30 min with 1 mL of 7.0% BF(3)/methanol produced a TT-TC/CT (21.54 mg) with the composition of 1.3% t12,t14; 5.9% t11,t13; 42.7% t10,t12; 44.0% t9,t11; 5.0% t8,t10; and 1.1% t 7,t9 CLA, whereas the methylation for 60 min with 14.0% BF(3)/methanol produced a TT-TC/CT (28.62 mg) with the composition of t,t CLA isomers different from that of TT-TC/CT by methylation for 30 min with 7.0% BF(3)/methanol. A large quantity of TT-TC/CT (14.15 g) with the composition similar to that of TT-TC/CT prepared from 50 mg of synthetic CLA was also prepared from 25 g of synthetic CLA. The purity of TT-TC/CT samples was greater than 98%. These results suggest that TT-TC/CT with a purity greater than 98% was easily prepared from synthetic CLA by BF(3)-catalyzed methylation, and the amount and composition of t,t CLA isomers of TT-TC/CT samples could be controlled by methylation conditions. PMID- 12744645 TI - A sensitive sandwich ELISA for the detection of trace amounts of cashew (Anacardium occidentale L.) nut in foods. AB - Trace amounts of cashew nut protein can provoke severe allergic reactions in sensitive patients. Consequently, commercial food processors and regulatory agencies must be vigilant to prevent cashew nut cross-contamination among foods and ensure proper labeling. Toward this end, we have developed a sandwich enzyme linked immunosorbent (ELISA) to detect the predominant cashew protein fraction (anacardein or cashew major protein, CMP) that can be extracted in aqueous buffer from food matrixes. Protein G-purified goat antiwhole cashew extract IgG and rabbit anti-CMP IgG were used as capture and secondary antibodies, respectively. Immunoadsorption against several nut and seed proteins significantly minimized the inherent cross-reactivity of these reagents. Food samples spiked with cashew flour and CMP were extracted and tested in a sandwich ELISA where standard curves were based on reactivity with CMP. The assay was optimized to detect as little as 20 ng/mL (0.02 ppm) of CMP and was successfully used to quantify CMP, and thus cashew, in various food matrixes. PMID- 12744646 TI - Determination of thiamine and its esters in beers and raw materials used for their manufacture by liquid chromatography with postcolumn derivatization. AB - Thiamine and its mono- and pyrophosphate esters were determined in beer and the raw materials used for its manufacture (brewer's yeast, malt, raw grain, and hops) after separation using reversed-phase liquid chromatography. The method used fluorescence detection and a new amide-based stationary phase, which avoids the need to form ion pairs, leading to narrower peaks and a simpler mobile phase. Analyses were performed by isocratic elution with a phosphate buffer mobile phase and using a postcolumn derivatization system based on the oxidation of thiamine to fluorescent thiochrome with potassium ferricyanide in alkaline solution. Only thiamine was found in the beers and raw products, especially in brewer's yeast and malt. A stability study pointed to a faster decrease in the thiamine content of samples stored at room temperature and in sunlight. PMID- 12744648 TI - Honey characterization and adulteration detection by pattern recognition applied on HPAEC-PAD profiles. 1. Honey floral species characterization. AB - An improved COFRAC (COmite FRancais d'ACreditation) method for the analysis and evaluation of the quality of honey by high-performance anion-exchange chromatography of sugar profiles is proposed. With this method, both minor and major sugars are simultaneously analyzed and the technique is integrated in a new chemometric approach, which uses the entire chromatographic sugars profile of each analyzed sample to characterize honey floral species. Sixty-eight authentic honey samples (6 varieties) were analyzed by high-performance anion-exchange chromatography-pulsed amperometric detection. A new algorithm was developed to create automatically the corresponding normalized data matrix, ready-to-use in various chemometric procedures. This algorithm transforms the analytical profiles to produce the corresponding calibrated table of the surfaces or intensities according to retention times of peaks. The possibility of taking into account unknown peaks (those for which no standards are available) allows the maximum chemical information provided by the chromatograms to be retained. The parallel application of principal component analysis (PCA)/linear discriminant analysis (LDA) and artificial neural networks (ANN) shows a high capability in the classification of the analyzed samples (LDA, 93%; ANN, 100%) and a very good discrimination of honey groups. This work is the starting point of the elaboration of a new system designed for the automatic pattern recognition of food samples (first application on honey samples) from chromatographic analyses for food characterization and adulteration detection. PMID- 12744647 TI - Development of enzyme-linked immunosorbent assays for isocupressic acid and serum metabolites of isocupressic acid. AB - The consumption of ponderosa pine (Pinus ponderosa), lodgepole pine (Pinus contorta), common juniper (Juniperus communis), and Monterey cypress (Cupressus macrocarpa) causes abortions in pregnant cattle. Recent studies have identified isocupressic acid (1) as the primary abortificient compound in these plants. In vitro and in vivo studies using rumen and blood have shown isocupressic acid (1) is rapidly metabolized to agathic acid (3), dihydroagathic acid (4), and tetrahydroagathic acid (5). Rapid and sensitive diagnostic techniques are needed to identify poisoned animals, to study toxicokinetics, and to elucidate the mechanism of isocupressic acid-induced abortion in cattle. In this study, four competitive inhibition enzyme-linked immunosorbent assays for isocupressic acid and its sera metabolites were developed using polyclonal antibodies. One assay is specific to 1, whereas the other three assays show cross-reactivity to 3-5 in addition to 1. The assay specific to 1 had a limit of detection of 44.1 pg. The other assays which demonstrated cross-reactivity to the isocupressic acid blood metabolites also had comparably low limits of detection. One assay was used to follow the absorption and elimination profile of isocupressic acid metabolites in both cow serum and urine after oral dosage of a cow with common juniper. PMID- 12744649 TI - Quantitative determination of sulfur-containing wine odorants at sub parts per billion levels. 2. Development and application of a stable isotope dilution assay. AB - [(2)H(10)]-4-Mercapto-4-methylpentan-2-one (d(10)-1), [(2)H(2)]-3-mercaptohexan-1 ol (d(2)-2), and [(2)H(5)]-3-mercaptohex-1-yl acetate (d(5)-3), deuterated analogues of impact odorants of wines, were used to determine quantitatively the natural compounds in white wines (Muscadet, Sauvignon, and Bacchus) with a stable isotope dilution assay using gas chromatography coupled either with ion trap tandem mass spectrometry (GC-ITMS-MS) or with atomic emission detection monitored on sulfur-selective acquisition (GC-AED). The thiol compounds were recovered from wines by liquid-liquid extraction, then purified from the wine extracts by covalent chromatography, and analyzed. The quantitative determination of 4 mercapto-4-methylpentan-2-one 1 in the wines that were analyzed was performed better with GC-AED than with GC-ITMS-MS under the conditions that were used. However, the detection limit of the method was higher than the odor threshold of 4-mercapto-4-methylpentan-2-one 1 in wine (5 vs 0.8 ng/L). The levels of this compound in the Sauvignon and Bacchus wines were much higher than its odor threshold, but it was not detectable in the Muscadet wines. On the contrary, GC ITMS-MS was much more sensitive than GC-AED for detection of 3-mercaptohexan-1-ol 2 and 3-mercaptohex-1-yl acetate 3, and the detection limits were much lower than their odor thresholds in wine. The former compound was detected in all of the Muscadet wines that were analyzed at levels always higher than its odor detection threshold, while the latter occurred at levels higher than its odor threshold in only one Muscadet wine. PMID- 12744650 TI - A rapid spectrofluorometric screening method for enrofloxacin in chicken muscle. AB - A simple spectrofluorometric method was developed for screening enrofloxacin (ENRO) in chicken muscle. A single-step extraction with acidic acetonitrile gave the best results without further cleanup. Following centrifugation the supernatants were excited at 324 nm and the emission was measured at 442 nm. Using this procedure, 18 chicken breast samples from 3 producers were tested. The results showed background signal levels significantly lower than those corresponding to 300 microg/kg ENRO, the FDA approved tolerance level. Statistical treatment of these data established a threshold which can be used in subsequent screening of ENRO at the tolerance level. The calibration curve revealed a satisfactory linear relationship (R(2) = 0.9991) in a range of 0-700 microg/kg ENRO in fortified chicken breast. ENRO-incurred samples were examined using this approach, and the results agreed with those obtained from more extensive separation followed by high-performance liquid chromatography. Because the threshold can be set at the 3 sigma limit, reliable screening can be accomplished with an error rate of less than 0.26%. Based on this investigation, a high-throughput screening method for ENRO in chicken tissue is proposed. PMID- 12744651 TI - Determination of albendazole and its major metabolites in the muscle tissues of Atlantic salmon, tilapia, and rainbow trout by high performance liquid chromatography with fluorometric detection. AB - A liquid chromatographic procedure for the determination of albendazole ([5 (propylthio)-1H-benzimidazol-2yl]carbamic acid methyl ester) and its major metabolites, albendazole sulfoxide, albendazole sulfone, and albendazole-2- aminosulfone in rainbow trout, tilapia, and salmon muscle with adhering skin tissue is described. The muscle tissue samples are made alkaline with potassium carbonate and extracted with ethyl acetate. The extracts are further subjected to cleanup by utilizing a number of liquid-liquid extraction steps. After solvent evaporation, the residue is reconstituted in mobile phase and chromatographed. The chromatography is carried out on a reversed phase Luna C(18) column, using acetonitrile/methanol/buffer as a mobile phase and a fluorescence detector. The average recoveries from the fortified muscle tissue of the three fish species for albendazole (25-100 ppb), albendazole sulfoxide (15.5-62 ppb), albendazole sulfone (1-10 ppb), and albendazole-2- aminosulfone (10-100 ppb) were 94, 77, 82, and 67%, respectively. The average CV for each compound was < or =10%. The procedure was validated and then applied to the determination of albendazole and its three major metabolites in the muscle tissue of the three fish species obtained after orally dosing with albendazole. PMID- 12744652 TI - Characterization of triterpene alcohol and sterol ferulates in rice bran using LC MS/MS. AB - Ferulic acid esters of triterpene alcohols and sterols in rice bran oil have been extensively studied and reported to possess important pharmacological actions. Inconsistent results on the numbers and structures of ferulates have been reported, primarily because of the analytical procedures employed. Conventional methods for analysis of phytosterol content in oil are carried out by characterization of trimethylsilylated derivatives (TMS) using GC-EI-MS after saponification of oils or individual compound isolated from oils. This study developed an LC-MS/MS method for the direct analysis of triterpene alcohol and sterol esters in rice bran oil. In addition to verifying the results of previous research, nine new relatively polar triterpene alcohol and sterol esters were characterized by their retention behaviors in LC and ESI-MS data from both negative- and positive-ion mode. This is the first evidence for the presence of hydroxylated ferulate esters and caffeate esters as part of gamma-oryzanol in rice bran. The method enables rapid and direct on-line characterization of triterpene alcohol and sterol esters in oils. LC-MS/MS equipped with reverse phase LC and ESI-MS should be well-suited for identification and quantification of the polar metabolites of phytosterols in biological fluids after consumption of rice bran oil or other oils. PMID- 12744653 TI - FTIR-ATR analysis of brewed coffee: effect of roasting conditions. AB - FTIR-ATR was used to study the effect of roasting conditions on the flavor of brewed coffee using Guatemala Antigua coffee beans. The 1800-1680 cm(-1) carbonyl region for vinyl esters/lactones, esters, aldehydes, ketones, and acids was found to provide a flavor-print of the brewed coffee. A study of light, medium, and dark roasts indicated that when the rate of heating to the onset of the first and second cracks was kept constant, the types of carbonyl compounds formed were similar, varying only in their concentration. This difference in concentration is apparently due to the additional heating of the coffee bean beyond the second crack. When the heating rate to the onset of the first and second crack was varied, both the types and concentration of the carbonyl compounds formed during roasting were affected. Thus, heating rates of green coffee beans to the onset of the first and second cracks are important determinants of the basic taste and aroma of brewed coffee. PMID- 12744654 TI - Assays for hydrophilic and lipophilic antioxidant capacity (oxygen radical absorbance capacity (ORAC(FL))) of plasma and other biological and food samples. AB - Methods are described for the extraction and analysis of hydrophilic and lipophilic antioxidants, using modifications of the oxygen radical absorbing capacity (ORAC(FL)) procedure. These methods provide, for the first time, the ability to obtain a measure of "total antioxidant capacity" in the protein free plasma, using the same peroxyl radical generator for both lipophilic and hydrophilic antioxidants. Separation of the lipophilic and hydrophilic antioxidant fractions from plasma was accomplished by extracting with hexane after adding water and ethanol to the plasma (hexane/plasma/ethanol/water, 4:1:2:1, v/v). Lipophilic and hydrophilic antioxidants were efficiently partitioned between hexane and aqueous solvents. Conditions for controlling temperature effects and decreasing assay variability using fluorescein as the fluorescent probe were validated in different laboratories. Incubation (37 degrees C for at least 30 min) of the buffer to which AAPH was dissolved was critical in decreasing assay variability. Lipophilic antioxidants represented 33.1 +/- 1.5 and 38.2 +/- 1.9% of the total antioxidant capacity of the protein free plasma in two independent studies of 6 and 10 subjects, respectively. Methods are described for application of the assay techniques to other types of biological and food samples. PMID- 12744655 TI - Capillary electrophoresis of alpha-lactalbumin in milk powders. AB - A quick and simple method for the extraction and analysis of alpha-lactalbumin in milk protein powders is presented, which permits accurate duty classification of commercial products as required under the U.S. Harmonized Tariff System. An acetic acid buffer medium is utilized to extract the whey proteins, which are then analyzed by capillary electrophoresis using an electrokinetic injection onto an uncoated column. The running buffer is composed of a 50 mM borate buffer at pH 8.0. The separation is rapid (<5 min) and displays good limits of detection (+/ 0.01 mg mL(-1)). Overall, the method provides a simple means for extracting and analyzing alpha-lactalbumin in milk powders. PMID- 12744656 TI - Purification and structural characterization of 3-hydroxypropionaldehyde and its derivatives. AB - The compound 3-hydroxypropionaldehyde (3-HPA), together with HPA hydrate and HPA dimer, in aqueous solution forms a system with interesting chemical properties. Therefore, 3-HPA has attracted attention by the chemical industry for use as a precursor in the production of plastics, acrylic acid, and 1,3-propanediol and by the food industry, in using 3-HPA-producing Lactobacillus reuteri as a probiotic. To produce 3-HPA in high yield from glycerol, L. reuteri was used as a biotransformation system. A convenient chromatographic purification method was developed, and purified 3-HPA was analyzed using electrospray ionization mass spectrometry and (13)C NMR. Quantitative (13)C NMR revealed a concentration dependent distribution of the three compounds forming the HPA system. At concentrations above 1.4 M, the HPA dimer was predominant. However, at concentrations relevant for biological systems, HPA hydrate was the most abundant, followed by the aldehyde form. Our results indicate that the dimeric form with expected antibiotic properties should not be the active form. PMID- 12744657 TI - Volatile metabolites from Salvia fruticosa as antifungal agents in soilborne pathogens. AB - The volatile metabolites of Salvia fruticosa plants, growing wild in 15 localities scattered across Greece, were analyzed by means of GC and GC-MS. The essential oil content ranged from 0.69 to 4.68%, and the results of the analyses showed a noticeable variation in the amounts of the five main components [1,8 cineole, alpha-thujone, beta-thujone, camphor, and (E)-caryophyllene]. The antifungal activities of the essential oils from two localities, belonging in two different groups of cluster and principal component analysis, and their main components (1,8-cineole and camphor) were evaluated in vitro against five phytopathogenic fungi. Both oils were slightly effective against Fusarium oxysporum f. sp. dianthi and Fusarium proliferatum, whereas against Rhizoctonia solani, Sclerotinia sclerotiorum, and Fusarium solani f. sp. cucurbitae the oils exhibited high antifungal activities. PMID- 12744658 TI - Antioxidative and hepatoprotective effects of Antrodia camphorata extract. AB - Antrodia camphorata (A. camphorata) is well-known in Taiwan as a traditional Chinese medicine. The purpose of this study was to evaluate the ability of A. camphorata extracts to protect against oxidative stress in vitro and against carbon tetrachloride (CCl(4))-induced hepatic injury in vivo. An extract of A. camphorata inhibited nonenzymatic iron-induced lipid peroxidation in rat brain homogenates with an IC(50) value about 3.1 mg/mL. It also scavenged the stable free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH). The dose of the A. camphorata extract resulting in a decrease of 0.20 in the absorbance of DPPH was about 31 +/ 0.7 microg/mL. Furthermore, an A. camphorata extract dose-dependently (250-1250 mg/kg) ameliorated the increase in plasma aspartate aminotransferase (GOT) and alanine aminotransferase (GPT) levels caused by chronic repeated CCl(4) intoxication in mice. Moreover, A. camphorata extract significantly improved the CCl(4)-induced increase in hepatic glutathione peroxidase, reductase, and CCl(4) induced decrease in superoxide dismutase activities. It also restored the decrement in the glutathione content and catalase activity of hepatic tissues in CCl(4)-intoxicated mice. Furthermore, it also dose-dependently inhibited the formation of lipid peroxidative products during CCl(4) treatment. Histopathological changes of hepatic lesions induced by CCl(4) were significantly ameliorated by treatment with an A. camphorata extract in a dose-dependent manner. These results suggest that A. camphorata extract exerts effective protection against chronic chemical-induced hepatic injury in vivo, by mediating antioxidative and free radical scavenging activities. PMID- 12744659 TI - Dihydrochalcones: evaluation as novel radical scavenging antioxidants. AB - Dihydrochalcones are a family of bicyclic flavonoids, defined by the presence of two benzene rings joined by a saturated three carbon bridge. In the present study, we systematically examined the antioxidant activities of dihydrochalcones against the stable free radical (1,1-diphenyl-2-picrylhydrazyl) and lipid peroxidation in the erythrocyte membrane. All dihydrochalcones exhibited higher antioxidant activities than the corresponding flavanones. The (1)H NMR analysis indicated that the active dihydrochalcone has a time-averaged conformation in which the aromatic A ring is orthogonal to the carbonyl group, while the inactive dihydrochalcone such as 2'-O-methyl-phloretin has a strongly hydrogen-bonded phenolic hydroxyl group, suggestive of a coplanar conformation. A hydroxyl group at the 2'-position of the dihydrochalcone A ring, newly formed by reduction of the flavanone C ring, is an essential pharmacophore for its radical scavenging potential. PMID- 12744660 TI - Stoichiometric and kinetic studies of phenolic antioxidants from Andean purple corn and red-fleshed sweetpotato. AB - Stoichiometric and kinetic values of phenolics against DPPH (2,2-diphenyl-1 picrylhydrazyl) were determined for Andean purple corn (Zea mays L.) and red sweetpotato (Ipomoea batatas). Both crops had higher antioxidant capacity and antiradical kinetics than blueberries and higher or similar anthocyanin and phenolic contents. The second-order rate constant (k(2)) was 1.56, 1.12, 0.57, and 0.26 (mg antiradical/mL)(-1) s(-1) for red sweetpotato, Trolox, purple corn, and blueberry, respectively. On the molar basis of active hydroxyl groups, k(2)' showed the same order as for k(2). Corn cob and sweetpotato endodermis contributed the most in phenolic compounds and antioxidant capacity. Both crops studied can be considered as excellent novel sources of natural antioxidants for the functional food and dietary supplement markets. PMID- 12744661 TI - Food matrix effects on bioactivity of broccoli-derived sulforaphane in liver and colon of F344 rats. AB - Sulforaphane (SF) is considered to be the major anticarcinogenic component in broccoli. The effects of feeding rats purified SF (5 mmol/kg of diet), broccoli containing SF formed in situ during laboratory hydrolysis (broccoli-HP; 20% freeze-dried broccoli diet, 0.16 mmol of SF/kg of diet), and broccoli containing intact glucosinolates (broccoli-GS; 20% freeze-dried broccoli diet, 2.2 mmol of glucoraphanin/kg of diet) were compared. Rats (male F344 rats, five per group) were fed control (modified AIN-76 B-40), SF, broccoli-HP, or broccoli-GS for 5 days. In rats fed broccoli-GS, quinone reductase activities (QR) in the colon and liver were greater (4.5- and 1.4-fold over control, respectively) than in rats fed broccoli-HP (3.2- and 1.1-fold over control, respectively). Broccoli-GS and SF diets increased QR to the same extent, even though the broccoli-GS diet contained far less SF (as the unhydrolyzed glucosinolate, glucoraphanin) than the purified SF diet. In a second experiment, rats were fed one of six diets for 5 days: (1) control; (2) 20% broccoli-GS; (3) diet 2 + low SF (0.16 mmol/kg of diet); (4) diet 2 + high SF (5 mmol/kg of diet); (5) low SF (0.32 mmol/kg of diet); or (6) high SF (5.16 mmol/kg of diet). In both liver and colon, QR was increased most by high SF plus broccoli-GS; individually, high SF and broccoli-GS had similar effects, and adding the low-dose SF to broccoli-GS had either no effect or a negative effect. In both experiments, urinary SF-mercapturic acid correlated with QR activity, not with dietary intake. It was concluded that all diets were substantially more effective in the colon than in the liver and that broccoli-GS was more potent than SF or broccoli-HP. PMID- 12744662 TI - Endophytic fungi associated with Mediterranean plants as a source of mycelium bound lipases. AB - A screening of endophytic fungi isolated from Mediterranean plants rendered a mycelium-bound lipase from a strain of Rhizopus oryzae that catalyzed the esterification of fatty acids in isooctane. The influence of various factors (water content, temperature, and pH) on ester synthesis was investigated. Catalytic activity was inversely correlated with water content. This enzyme was active over the entire pH range studied, from pH 3 to pH 8, and activity was maximal at pH 4 and pH 7. The enzyme was thermostable, with maximal activity at 60 degrees C. PMID- 12744663 TI - Protection against Trp-P-2 DNA adduct formation in C57bl6 mice by purpurin is accompanied by induction of cytochrome P450. AB - Purpurin, an anthraquinone constituent from madder root, has previously been reported as antimutagenic in the Ames Salmonella bacterial mutagenicity assay and as antigenotoxic in Drosophila melanogaster, against a range of environmental carcinogens. Short-term dietary supplementation with purpurin inhibits the formation of hepatic DNA adducts in male C57bl6 mice after a single dose of the heterocyclic amine dietary carcinogen Trp-P-2 (30 mg/kg). Inhibition of adduct formation was dose-dependent. No DNA adducts were observed in animals treated only with purpurin. The decrease in adduct formation was accompanied by significant, dose-dependent inductions of hepatic cytochrome P450-dependent dealkylations of methoxy- (CYP1A2), ethoxy- (CYP1A1), and pentoxy- (CYP2B) resorufins, total cytochrome P450, and NADPH cytochrome P450 reductase. It is hypothesized that purpurin exhibits chemopreventive potential by inhibiting the cytochrome P450-dependent metabolism of heterocyclic amines to their genotoxic N hydroxylamines. PMID- 12744664 TI - Commercial scale pulsed electric field processing of tomato juice. AB - Effects of commercial scale pulsed electric field (PEF) processing on the quality of tomato juice were studied and compared with those of thermal processing. Tomato juice was prepared by hot break at 88 degrees C for 2 min or by cold break at 68 degrees C for 2 min and then thermally processed at 92 degrees C for 90 s or PEF processed at 40 kV/cm for 57 micros. Thermally processed, PEF processed, and unprocessed control juices were packed into 50 mL sterilized polypropylene tubes in a sanitary glovebox and stored at 4 degrees C for 112 days. Both thermally and PEF processed juices showed microbial shelf life at 4 degrees C for 112 days. The lipoxygenase activities of thermally and PEF processed juices were 0 and 47%, respectively. PEF processed juice retained more ascorbic acid than thermally processed juice at 4 degrees C for 42 days (p < 0.05). No significant differences were observed in the concentration of lycopene, degrees Brix, pH, or viscosity between thermally and PEF processed juices during the storage (p > 0.05). Sensory evaluations indicated that flavor and overall acceptability of PEF processed juice were preferred to those of thermally processed juice (p < 0.05). PMID- 12744665 TI - Chemometric studies of vinegars from different raw materials and processes of production. AB - The phenolic composition, aroma compounds, and organic acid content of 83 vinegars have been determined. Multivariate analysis techniques have been used to classify these vinegar samples according to raw material (white wine, red wine, apple, honey, alcohol, balsamic, and malt) and production process (with and without aging in wood). Cluster analysis grouped the samples according to production process. Only apple and balsamic vinegars were separated from wine vinegars. Alcohol, honey, and malt vinegars were grouped with no aged wine vinegars. Linear discriminate analysis allowed a 88% differentiation according to raw material and 100% according to aging in wood. Besides, from the results obtained, a major role of the volatile compounds in the differentiation of the vinegar samples according to their aging period in wood can be seen. PMID- 12744666 TI - Model studies on the stability of folic acid and 5-methyltetrahydrofolic acid degradation during thermal treatment in combination with high hydrostatic pressure. AB - Stability of folic acid and 5-methyltetrahydrofolic acid in phosphate buffer (0.2 M; pH 7) toward thermal (above 65 degrees C) and combined high pressure (up to 800 MPa)/thermal (20 up to 65 degrees C) treatments was studied on a kinetic basis. Residual folate concentration after thermal and high pressure/thermal treatments was measured using reverse phase liquid chromatography. The degradation of both folates followed first-order reaction kinetics. At ambient pressure, the estimated Arrhenius activation energy (E(a)) values of folic acid and 5-methyltetrahydrofolic acid thermal degradation were 51.66 and 79.98 kJ mol( 1), respectively. It was noticed that the stability of folic acid toward thermal and combined high pressure thermal treatments was much higher than 5 methyltetrahydrofolic acid. High-pressure treatments at room temperature or higher (up to 60 degrees C) had no or little effect on folic acid. In the whole P/T area studied, the rate constant of 5-methyltetrahydrofolic acid degradation was enhanced by increasing pressure, and a remarkable synergistic effect of pressure and temperature on 5-methyltetrahydrofolic acid degradation occurred at temperatures above 40 degrees C. A model to describe the combined pressure and temperature effect on the 5-methyltetrahydrofolic acid degradation rate constant is presented. PMID- 12744667 TI - Thermal decomposition of specifically phosphorylated D-glucoses and their role in the control of the Maillard reaction. AB - One of the main shortcomings of the information available on the Maillard reaction is the lack of knowledge to control the different pathways, especially when it is desired to direct the reaction away from the formation of carcinogenic and other toxic substances to more aroma and color generation. The use of specifically phosphorylated sugars may impart some elements of control over the aroma profile generated by the Maillard reaction. Thermal decomposition of 1- and 6-phosphorylated glucoses was studied in the presence and absence of ammonia and selected amino acids through pyrolysis/gas chromatography/mass spectrometry using nonpolar PLOT and medium polar DB-1 columns. The analysis of the data has indicated that glucose-1-phosphate relative to glucose undergoes more extensive phosphate-catalyzed ring opening followed by formation of sugar-derived reactive intermediates as was indicated by a 9-fold increase in the amount of trimethylpyrazine and a 5-fold increase in the amount of 2,3-dimethylpyrazine, when pyrolyzed in the presence of glycine. In addition, glucose-1-phosphate alone generated a 6-fold excess of acetol as compared to glucose. On the other hand, glucose-6-phosphate enhanced retro-aldol reactions initiated from a C-6 hydroxyl group and increased the subsequent formation of furfural and 4-cyclopentene-1,3 dione. Furthermore, it also stabilized 1- and 3-deoxyglucosone intermediates and enhanced the formation of six carbon atom-containing Maillard products derived directly from them through elimination reactions such as 1,6-dimethyl-2,4 dihydroxy-3-(2H)-furanone (acetylformoin), 2-acetylpyrrole, 5-methylfurfural, 5 hydroxymethylfurfural, and 4-hydroxy-2,5-dimethyl-3-(2H)-furanone (Furaneol), due to the enhanced leaving group ability of the phosphate moiety at the C-6 carbon. However, Maillard products generated through the nucleophilic action of the C-6 hydroxyl group such as 2-acetylfuran and 2,3-dihydro-3,5-dihydroxy-4H-pyran-4-one were retarded, due to the blocked nucleophilic atom at C-6. PMID- 12744668 TI - Volatile emissions of navel oranges as predictors of freeze damage. AB - Volatile emissions of navel orange (Citrus sinensis L. Osbeck cv. Washington) fruit were evaluated as a means for predicting and gauging freeze damage. The fruits were subjected to -5 or -7 degrees C treatments in a laboratory freezer for various time periods of 2-9.5 h and stored at 23 degrees C for 1, 2, or 7 days, after which time the emission of volatiles from the fruit was measured. Following the final day of volatile measurements the fruits were stored at 5 degrees C for an additional 2-3 weeks and then evaluated for fruit quality characteristics. Peel injury in the form of brown lesions, drying of the juice vesicles, a decline in acidity, and a loss of flavor were observed to occur as a result of freezing. Corresponding to the loss in fruit quality were large increases in the emissions of ethanol, ethyl butanoate, methyl hexanoate, and ethyl octanoate. With the exception of methyl hexanoate, for which volatile emissions decreased during storage for 7 days at 23 degrees C, all of the other volatiles were relatively unchanged in amount by storage. Treatment at -7 degrees C caused greater injury, quality loss, and more volatile emanation than did freezing at -5 degrees C. The measurement of volatile emissions appears to be a useful approach to identify freeze-damaged navel oranges. PMID- 12744669 TI - Anthocyanin composition of Cabernet Sauvignon and Tempranillo grapes at different stages of ripening. AB - Changes in anthocyanins during ripening of Cabernet Sauvignon and Tempranillo grapes were studied over a three year period. The accumulation of anthocyanins showed variations during ripening, especially during the first three weeks after veraison, and the accumulation pattern of those molecules changed only slightly from one year to another. On the other hand, the percentages of the different anthocyanins studied were different for each cultivar, and some changes were observed in both cultivars depending on the weather conditions of the growing season. In warm years the percentages of primitive anthocyanins (delphinidin 3-O glucoside and petunidin 3-O-glucoside) were slightly lower than in a relatively cool year. Nevertheless, the anthocyanin fingerprints of Cabernet Sauvignon and Tempranillo grapes seem to be rather stable during ripening, despite the sugar content of the grapes. PMID- 12744670 TI - Effect of selenium on the yield and quality of green tea leaves harvested in early spring. AB - Foliar applications of a fertilizer of selenite or selenate were carried out to determine the influence of selenium on the yield and quality of green tea leaves harvested in early spring. Numbers of sprouts and the yield were significantly increased by the application of selenium. The sweetness and aroma of green tea leaves were also significantly enhanced, and bitterness was significantly decreased by the application of selenium. However, no significant differences were found in sweetness, bitterness, and aroma between tea leaves fertilized with selenite and selenate. Se concentration was significantly increased by selenium fertilization, and tea enriched by sodium selenate had a significantly higher selenium content than did tea enriched by sodium selenite. Total amino acid and vitamin C contents were significantly enhanced by the application of selenium. Tea polyphenol contents were significantly decreased by fertilization with selenium. The marked difference of tea polyphenols was also found between applications of selenite and selenate. PMID- 12744671 TI - Variation in isoflavone of soybean cultivars with location and storage duration. AB - Fifteen soybean [Glycine max (L.) Merrill] cultivars were grown in Seoul, Suwon, and Kyongsan, Korea, in 1998, 1999, and 2000, and their isoflavone contents were assessed. After harvest, the beans were stored for 3 years at room temperature. Soybean isoflavones were analyzed using high-performance liquid chromatography (HPLC) within each crop year and after storage. Total isoflavone contents ranged from 188.4 to 685.6 mg 100 g(-1) in 1998, from 218.8 to 948.9 mg 100 g(-1) in 1999, and from 293.1 to 483.0 mg 100 g(-1) in 2000. The year x variety, and year x location x variety interactions were significantly different in 1998, the year x location, year x variety, and year x location x variety interactions were significantly different in 1999, and the year x variety interaction was significantly different in 2000 for total and individual isoflavone contents. Total isoflavone contents of soybeans stored for 1 year were only slightly higher than those of soybeans stored for 2 or 3 years. However, the concentrations of individual isoflavones, especially 6' '-O-malonyldaidzin and 6' '-O malonylgenistin, decreased markedly in soybeans stored for 2 or 3 years. These data suggest that it may be feasible to improve soybean cultivars with higher antioxidative substances. PMID- 12744672 TI - Kinetics for substrate utilization and methane production during the mesophilic anaerobic digestion of two phases olive pomace (TPOP). AB - A kinetic study of the anaerobic digestion process of two phases olive pomace (TPOP) was carried out in a laboratory-scale completely stirred tank reactor at mesophilic temperature (35 degrees C). The reactor was operated at influent substrate concentrations of 34.5 (substrate I), 81.1 (substrate II), 113.1 (substrate III), and 150.3 g COD/L (substrate IV). The hydraulic retention times (HRT) ranged between 8.3 and 40.0 days for the most diluted substrate (I) and between 10 and 50 days for the other three influent substrate concentrations used (substrates II-IV). The results obtained demonstrated that the rates of substrate uptake and methane production were correlated with the concentration of biodegradable total chemical oxygen demand (COD), through equations of the Michaelis-Menten type. A mass (COD) balance around the reactor allowed the methane yield coefficient and cell maintenance coefficient to be obtained, which gave values of 0.25 L CH(4)/g COD(t) and 0.25 days(-1), respectively. The first one was coincident to that obtained through experimental data of methane production and substrate consumption. The kinetic equations obtained and the proposed mass balance were used to simulate the anaerobic digestion process of TPOP and to obtain the theoretical COD of the reactor and methane production rates. The small deviations obtained (equal or lower than 10%) between the values calculated through the model and experimental ones suggest that the proposed model predicts the behavior of the reactor very accurately. PMID- 12744673 TI - Dissipation of sulfosulfuron in soil and wheat plant under predominant cropping conditions and in a simulated model ecosystem. AB - Environmental fate and dissipation of the sulfonylurea herbicide sulfosulfuron was investigated in soil (inceptisol) and wheat plant under predominant cropping conditions. Studies were conducted in natural field conditions and in a simulated model ecosystem. Thirty days after the wheat seeds had been sown, sulfosulfuron [N-[[(4,6-dimethoxy-2-pyrimidinyl)amino]carbonyl]-2-(ethylsulfonyl)imidazo[1,2 a]pyridine-3-sulfonamide] 75% w/w WG formulation was applied once in the field. The dosages were 25 and 50 g of active ingredient (ai)/ha. Studies were conducted in significantly separated individual plots to avoid contamination. In a predetermined interval, soil samples were collected and analyzed for the residues of sulfosulfuron. At harvest, wheat grain, straw, and soil samples were analyzed for the residues. Similar experiments were conducted in a model ecosystem. Apart from this, after harvest, the succeeding crops coriander (Coriandrum sativum) and edible amaranth (Amaranthus mangostanus L.) were raised in the model ecosystem and studied for the residues. No residues were detected in wheat grain, straw, and soil samples collected at harvest from both experiments or in the succeeding crops coriander and edible amaranth in the model ecosystem when tested at the minimum detection level of 0.001 microg/g. The dissipation of sulfosulfuron was found to have first-order kinetics in soil and plant in both studies. The dissipation data of sulfosulfuron in the model ecosystem were compared with those from the natural field conditions. PMID- 12744674 TI - Predicting regional emissions and near-field air concentrations of soil fumigants using modest numerical algorithms: a case study using 1,3-dichloropropene. AB - Soil fumigants, used to control nematodes and crop disease, can volatilize from the soil application zone and into the atmosphere to create the potential for human inhalation exposure. An objective for this work is to illustrate the ability of simple numerical models to correctly predict pesticide volatilization rates from agricultural fields and to expand emission predictions to nearby air concentrations for use in the exposure component of a risk assessment. This work focuses on a numerical system using two U.S. EPA models (PRZM3 and ISCST3) to predict regional volatilization and nearby air concentrations for the soil fumigant 1,3-dichloropropene. New approaches deal with links to regional databases, seamless coupling of emission and dispersion models, incorporation of Monte Carlo sampling techniques to account for parametric uncertainty, and model input sensitivity analysis. Predicted volatility flux profiles of 1,3 dichloropropene (1,3-D) from soil for tarped and untarped fields were compared against field data and used as source terms for ISCST3. PRZM3 can successfully estimate correct order of magnitude regional soil volatilization losses of 1,3-D when representative regional input parameters are used (soil, weather, chemical, and management practices). Estimated 1,3-D emission losses and resulting air concentrations were investigated for five geographically diverse regions. Air concentrations (15-day averages) are compared with the current U.S. EPA's criteria for human exposure and risk assessment to determine appropriate setback distances from treated fields. Sensitive input parameters for volatility losses were functions of the region being simulated. PMID- 12744675 TI - Sulfentrazone adsorbed on micelle-montmorillonite complexes for slow release in soil. AB - Interactions of the herbicide sulfentrazone with the cationic surfactants octadecyltrimethylammonium (ODTMA), hexadecyltrimethylammonium (HDTMA), and benzyldimethylhexadecylammonium (BDMHDA) have been studied for the design of slow release formulations based on sulfentrazone adsorbed on a micelle-montmorillonite complex. Adsorbed amounts of sulfentrazone on ODTMA- and BDMHDA-montmorillonite complexes were 99.2-99.8% of that added, and desorption of herbicide in water during 24 h was low. After 10 washings in funnels with soil, only 2.6% of herbicide was released from ODTMA-montmorillonite formulations versus 100% release from the commercial formulation. The strong binding of sulfentrazone to micelles was confirmed by pH and spectroscopic measurements and was explained by the formation of ionic pairs between cationic surfactant and anionic herbicide. The ODTMA-clay and commercial formulations of sulfentrazone yield almost complete and 40% growth inhibition of green foxtail, respectively, at 700 g of active ingredient/ha. Hence, the slow release from micelle-clay formulations of sulfentrazone promotes its biological activity and reduces environmental contamination. PMID- 12744676 TI - Evaluation of aroma compounds contributing to muskmelon flavor in Porapak Q extracts by aroma extract dilution analysis. AB - The flavor of the Miyabi variety of Japanese muskmelon was extracted according to the Porapak Q column method (PQM) and evaluated by using aroma extract dilution analysis (AEDA) method. The overall odor of the PQM extracts was perceived as having a natural muskmelon-like odor, suggesting that the PQM was able to extract volatile compounds in muskmelon fruit without degradation of original flavor. Forty-six odorant compounds [Kovats index (KI), 961 < or = KI < or = 2605] were found by GC-sniffing in PQM extracts, confirming the effectiveness of PQM in trapping a wide range of volatile compounds in muskmelon flavor. The 46 odorants could be divided into three groups on the basis of their odor attributes: fruity note (KI < 1300); green, grassy, or cucumber-like note (1300 < KI < 2020); and sweet note (KI > 2020). When the original extracts were diluted in AEDA analysis, seven odorants could still be detected by GC-sniffing at a flavor diluation (FD) factor of 128 or above: one had a fruity note (compound 3); four had a cucumber like, green, or grassy note (compounds 12, 17, 21, and 23); and two had a sweet note (caramel-like or yakitori-like) (compounds 32 and 34). PMID- 12744677 TI - Impact odorants of different young white wines from the Canary Islands. AB - Five young monovarietal white wines from the Canary Islands made from Gual, Verdello, Marmajuelo, white Listan, and Malvasia grape cultivars were studied to determine the characteristics of their most important aromas and the differences among them. The study was carried out using gas chromatography-olfactometry (GC O) to detect the potentially most important aroma compounds, which were then analyzed quantitatively by gas chromatography-flame ionization detection and gas chromatography-mass spectrometry. The strongest odorants in the GC-O experiments were similar in all cases, although significant differences in intensity between samples were noted. Calculation of the odor activity values (OAVs) showed that 3 mercaptohexyl acetate was the most active odorant in the Marmajuelo and Verdello wines, as were 3-methylbutyl acetate in the Gual wine, beta-damascenone in the Malvasia wine, and ethyl octanoate in the white Listan wine. However, the most important differences between varieties were caused by the three mercaptans (3 mercaptohexyl acetate, 3-mercaptohexanol, and 4-methyl-4-mercapto-2-pentanone) and the vinylphenols (4-vinylphenol and 2-methoxy-4-vinylphenol). The correlation between the olfactometric values and the OAVs was satisfactory in the cases when the compound eluted in the GC-O system was well isolated from other odorants and had aromatic importance and the OAVs for the different wines were sufficiently different. PMID- 12744678 TI - Aroma compounds formed from 3-methyl-2,4-nonanedione under photooxidative conditions. AB - The behavior of the prominent aroma compound 3-methyl-2,4-nonanedione under photooxidative conditions was investigated in a model experiment. The four well known aroma compounds 2,3-butanedione, 2,3-octanedione, acetic acid, and caproic acid were identified. The main oxidation product was 3-hydroxy-3-methyl-2,4 nonanedione, an aroma compound with the odor description of rubbery, earthy, and plastic-like (GC-O). Its structure has been tentatively assigned based on mass (GC-MS) and vapor phase infrared spectra (GC-IR). The formal formation pathways are discussed for these compounds, and other origins described in the literature are presented. PMID- 12744680 TI - Aroma extract dilution analysis of Cv. Marion (Rubus spp. hyb) and Cv. Evergreen (R. laciniatus L.) blackberries. AB - Cultivar Marion and Evergreen blackberry aromas were analyzed by aroma extract dilution analysis. Sixty-three aromas were identified (some tentatively) by mass spectrometry and gas chromatography-retention time; 48 were common to both cultivars, and 27 have not been previously reported in blackberry fruit. A comparison of cultivars shows that both have comparable compound types and numbers but with widely differing aroma impacts, as measured by flavor dilution (FD) factors. Ethyl 2-methylbutanoate, ethyl 2-methylpropanoate, hexanal, furanones (2,5-dimethyl-4-hydroxy-3-(2H)-furanone, 2-ethyl-4-hydroxy-5-methyl-3 (2H)-furanone, 4-hydroxy-5-methyl-3-(2H)-furanone, 4,5-dimethyl-3-hydroxy-2-(5H) furanone, and 5-ethyl-3-hydroxy-4-methyl-2-(5H)-furanone), and sulfur compounds (thiophene, dimethyl sulfide, dimethyl disulfide, dimethyl trisulfide, 2 methylthiophene, and methional) were prominent in Evergreen (FD 512-2048). Except for ethyl 2-methylpropanoate, these same compounds were also prominent in Marion, but the FD factors varied significantly (FD 8-256) from Evergreen. The aroma profile of blackberry is complex, as no single volatile was unanimously described as characteristically blackberry. PMID- 12744679 TI - Analysis of volatiles in meat from Iberian pigs and lean pigs after refrigeration and cooking by using SPME-GC-MS. AB - The volatile compounds generated in meat from Iberian and lean pigs after four different treatments (raw, refrigerated, cooked, and refrigerated cooked meat) were analyzed. The different treatments showed different volatile profiles. Methyl alcohols and ketones (such as 2-ethyl-hexan-1-ol, 2-methyl-butan-1-ol, 3 methyl-butan-1-ol, and 3-hydroxy-butan-2-one) were the most representative in refrigerated meat because of the degradation of carbohydrates and proteins together with the Strecker degradation pathway. Lipid-derived volatiles were the most abundant in cooked meat and refrigerated cooked meat. Meat from different pig breeds presented different volatile profiles, probably due to different enzymatic and oxidative deterioration susceptibility. Otherwise, the fat content and its compositional characteristics also played an important role in the generation of volatiles. As compared to samples from lean pigs, muscles from Iberian pigs showed a higher content of heme iron that may have promoted the generation of higher content of total lipid-derived volatiles during the refrigeration of cooked meat. Despite that, the formation of volatiles with low thresholds and related to intense rancidity perception likely to be derived from polyunsaturated fatty acids was higher in lean pork than in meat from Iberian pigs. This might be expected to lead to a more intense development of a warmed over flavor during refrigeration of cooked samples from lean pigs. PMID- 12744682 TI - Distribution of hydroxycinnamic acid conjugates in fruit of commercial eggplant (Solanum melongena L.) cultivars. AB - There is gathering evidence that antioxidant phytonutrients in fruits and vegetables have health-promoting effects. Eggplant fruit have a high content of antioxidant phenolic compounds. We evaluated the main class of eggplant phenolics, hydroxycinnamic acid conjugates, in the fruit of seven commercial cultivars. Fourteen conjugates were quantified and identified by high-performance liquid chromatography, ES(-)-MS, and (1)H NMR data. Significant differences in their content and composition were evident among cultivars and in tissue from stem, middle, and blossom end segments. Chlorogenic acid (5-O-caffeoylquinic acid) was the predominant compound, and its 3-O-, 4-O-, and 5-O-cis isomers were also present. The 10 other phenolics fell into four groups, including 3,5- and 4,5-dicaffeoylquinic acid isomers, four amide conjugates, two unknown caffeic acid conjugates, and 3-O-acetyl esters of 5-O- and 4-O-caffeoylquinic acid. Dicaffeoylquinic and 3-O-acetyl chlorogenic acids were most variable among the cultivars. Dicaffeoyquinic acids were most abundant in the blossom end, whereas 3 O-acetyl esters were highest in the midsection. PMID- 12744681 TI - Distribution of volatile compounds in the pulp, cloud, and serum of freshly squeezed orange juice. AB - The quantitative distribution of volatile compounds in the pulp, cloud, and serum of a freshly squeezed orange juice (cv. Naveline) was measured. Juice monoterpene and sesquiterpene hydrocarbons were primarily recovered from the pulp (74.0 and 87.2%, respectively) and cloud (7.3 and 14.9%, respectively). Esters and monoterpene alcohols were mainly found in the serum (90.4 and 84.1%, respectively). Long chain aliphatic aldehydes tend to concentrate in the pulp. The relative proportions of individual volatile compounds were similar in the pulp and cloud. Pulp and cloud alcohol insoluble residues exhibited similar compositions; half of them are made of nonwall proteins, and the rest are made of cell wall materials. Pulp and cloud total and neutral lipids had similar fatty acids distributions, although the cloud was much richer in total lipids than the pulp. No relationship was found between the retention of aroma compounds in the pulp or cloud and their AIR and lipid content or composition. PMID- 12744683 TI - The effect of SO2 on the production of ethanol, acetaldehyde, organic acids, and flavor volatiles during industrial cider fermentation. AB - SO(2) is widely used in cider fermentation but also in other alcoholic beverages such as wine. Although the authorized limit is 200 ppm total SO(2), the International Organizations recommend its total elimination or at least reduction due to health concerns. Addition of SO(2) to apple juice at levels frequently used in industrial cidermaking (100 mg/L) induced significantly higher acetaldehyde production by yeast than that obtained without SO(2). Although the practical implications of acetaldehyde evolution under cidermaking conditions has been overcome by research and few data are available, this compound reached levels in two 2000 L bioreactors that may have prevented the occurrence of simultaneous alcoholic and malolactic fermentation. It was observed that malolactic fermentation had a positive effect promoting reduction of acetaldehyde levels in cider fermented with juice, SO(2)-treated or not. The addition of SO(2) clearly delayed malolactic fermentation comparing to the control, affecting not the onset of the malolactic fermentation but the rate of malic acid degradation. This compound, however, had a stimulatory effect on alcoholic fermentation. PMID- 12744684 TI - Antiphoto-oxidative activity of sesamol in methylene blue- and chlorophyll sensitized photo-oxidation of oil. AB - The effects and mechanism of sesamol on the methylene blue- or chlorophyll sensitized photo-oxidations of soybean oil have been studied. Sesamol showed strong antiphoto-oxidative activity in both methylene blue-and chlorophyll sensitized photo-oxidations of soybean oil in a dose-dependent manner. The 1.0 x 10(-3) M sesamol treatments showed 84.7 and 43.4% inhibitions of methylene blue- and chlorophyll-sensitized photo-oxidations of soybean oil in methylene chloride. The antiphoto-oxidative activity of sesamol was comparable to that of delta tocopherol in both methylene blue- and chlorophyll-sensitized photo-oxidations, at the same molar basis. Sesamol effectively inhibited rubrene oxidation with a chemical source of singlet oxygen in microemulsion, showing its strong singlet oxygen quenching ability. The results suggested that the antiphoto-oxidative activity of sesamol in the photo-oxidation of oil was, at least in part, due to its singlet oxygen scavenging activity. The singlet oxygen quenching rate constant (k(ox-Q) + k(q)) of sesamol was determined to be 1.9 +/- 0.3 x 10(7) M( 1) s(-1). This represents the first report on the antiphoto-oxidative activity of sesamol in the sensitized photo-oxidation of oil, and its bimolecular singlet oxygen quenching ability. PMID- 12744685 TI - Phaseolin in vitro pepsin digestibility: role of acids and phenolic compounds. AB - Great Northern bean (Phaseolus vulgaris L.) phaseolin proteolysis at 37 degrees C, varying HCl concentrations (10 mM to 1 M), phaseolin:pepsin ratios ranging from 5:1 to 100:1 (w/w), and incubation times up to 24 h was investigated. The results suggest that phaseolin is not resistant to in vitro pepsin hydrolysis. At a phaseolin-to-pepsin ratio of 100:1 (w/w), native phaseolin was completely digested in 24 h when incubated in 50 mM HCl, while heat-denatured phaseolin (30 min at 100 degrees C, boiling water bath) was digested in 1 h under similar conditions. When incubated at 37 degrees C for 24 h, acid alone, even at as low a concentration as 10 mM, caused a partial breakdown of native phaseolin. The degree of phaseolin hydrolysis by HCl was dependent on the acid concentration used. The rate of native phaseolin hydrolysis increased with increasing HCl concentration rather than pepsin concentration. Common food acids were able to partially hydrolyze phaseolin. Among the food acids tested, oxalic acid was the most effective in hydrolyzing phaseolin. Spectroscopic studies revealed a significant change in secondary and tertiary structures when native phaseolin was incubated in dilute HCl. None of the tested phenolic compounds adversely affected phaseolin hydrolysis by pepsin. PMID- 12744686 TI - Assessment of egg nutrient compositional changes and residue in eggs, tissues, and excreta following oral administration of atorvastatin to laying hens. AB - Laying hens were fed a control diet alone or with 0.06 g of atorvastatin, a synthetic 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, per 100 g of diet for 20 days. Compared to controls, egg yolks from treated hens contained greater amounts of amino acids and reduced levels of total fatty acids and cholesterol. In contrast, egg albumen amino acid contents were unaffected by dietary treatments. In a residue study, seven hens each received a single oral dose of approximately 20 microCi of [(14)C]atorvastatin. Approximately 71% of the radioactivity was recovered in the excreta and liver, whereas virtually no radioactivity was detected in kidney, heart, muscle, bile, plasma, or egg albumen at 15 days postdosing. Yolk radioactivity peaked at 4 days postdosing in six of the seven birds and was absent in eggs laid after day 10. Reminiscent of that of certain antibiotic drugs, the atorvastatin egg residue pattern appeared to coincide with the physiological pattern of daily yolk accretion within the ovary. PMID- 12744687 TI - Substrate suitability of different genotypes of sorghum in relation to Aspergillus infection and aflatoxin production. AB - Grain sorghum is often damaged by rain in the field and severely infected by grain mold, which includes Aspergillus infection and aflatoxin production. The objective of the study is to investigate the extent of aflatoxin production with Aspergillus infection in vitro in different sorghum genotypes with different pericarps, red, yellow, and white, the physical and chemical characteristics of grain during infection, and the changes in grain polyphenols and phytic acid in comparison to maize and groundnut. The physical characters and biochemical composition of sorghum grain contribute to make it less susceptible to Aspergillus infection and aflatoxin contamination compared to maize and groundnut. The lowest amounts of aflatoxin and ergosterol were observed in genotypes with red pericarp, whereas higher amounts of aflatoxin and ergosterol were found in white genotypes followed by maize and groundnut. All of the red genotypes differ in polyphenol composition and aflatoxin produced, showing resistance to mold damage. Another indication of resistance in red genotypes was the delayed peaking of aflatoxin production (9 days after infection). In red sorghum genotypes there was a significant, positive correlation existing between polyphenol content and aflatoxin produced at 3 and 6 days after infection, the r values being 0.589 and 0.513, respectively. The starch content decreased whereas the protein content in all sorghum genotypes increased during infection. Maximum phytic acid was observed in white sorghum genotypes. Phytic acid in yellow genotypes was found to have a significant negative correlation (r = -0.569) with aflatoxin produced. PMID- 12744688 TI - Repeated oral administration of high doses of the pomegranate ellagitannin punicalagin to rats for 37 days is not toxic. AB - The water-soluble ellagitanin punicalagin has been reported to be toxic to cattle. Taking into account that this antioxidant polyphenol is very abundant in pomegranate juice (> or =2 g/L), the present study evaluated the possible toxic effect of punicalagin in Sprague-Dawley rats upon repeated oral administration of a 6% punicalagin-containing diet for 37 days. Punicalagin and related metabolites were identified by HPLC-DAD-MS-MS in plasma, liver, and kidney. Five punicalagin related metabolites were detected in liver and kidney, that is, two ellagic acid derivatives, gallagic acid, 3,8-dihydroxy-6H-dibenzo[b,d]pyran-6-one glucuronide, and 3,8,10-trihydroxy-6H-dibenzo[b,d]pyran-6-one. Feedstuff intake, food utility index, and growth rate were lower in treated rats during the first 15 days without significant adverse effects, which could be due to the lower nutritional value of the punicalagin-enriched diet together with a decrease in its palatability (lower food intake). No significant differences were found in treated rats in any blood parameter analyzed (including the antioxidant enzymes gluthatione peroxidase and superoxide dismutase) with the exception of urea and triglycerides, which remained at low values throughout the experiment. Although the reason for the decrease is unclear, it could be due to the lower nutritional value of the punicalagin-enriched diet with respect to the standard rat food. Histopathological analysis of liver and kidney corroborated the absence of toxicity. In principle, the results reported here, together with the large safety margin considered, indicate the lack of toxic effect of punicalagin in rats during the 37 day period investigated. However, taking into account the high punicalagin content of pomegranate-derived foodstuffs, safety evaluation should be also carried out in humans with a lower dose and during a longer period of intake. PMID- 12744689 TI - Fluorescence of dietary porphyrins as a basis for real-time detection of fecal contamination on meat. AB - Digestion of green plants in the gastrointestinal (GI) tract produces degradation products from chlorophyll that cause ingesta and feces to be highly fluorescent. This property was exploited for development and construction of instruments to noninvasively detect minute quantities of feces on meat samples in real time. The presence of feces on meat products is a primary source of foodborne pathogens, such as Escherichia coli O157:H7 and Salmonella. This new technology provides a rapid and accurate alternative to the practice of visual inspection and augments more time-consuming biological testing methods. This innovation can assist meat processors and government inspectors in their efforts to provide safe and wholesome food to consumers. PMID- 12744691 TI - Synthesis, structure, and bonding in polyiodide and metal iodide-iodine systems. PMID- 12744692 TI - Physical mechanisms of generation and deactivation of singlet oxygen. PMID- 12744693 TI - Cyclometalated phosphine-based pincer complexes: mechanistic insight in catalysis, coordination, and bond activation. PMID- 12744694 TI - Conceptual density functional theory. PMID- 12744695 TI - Palladium-assisted routes to nucleosides. PMID- 12744696 TI - Vibrational spectroscopic detection of beta- and gamma-turns in synthetic and natural peptides and proteins. PMID- 12744697 TI - Properties and production of valienamine and its related analogues. PMID- 12744699 TI - Enantiomeric steroids: synthesis, physical, and biological properties. PMID- 12744698 TI - Palladium-catalyzed alkynylation. PMID- 12744700 TI - Unsaturated nitriles: conjugate additions of carbon nucleophiles to a recalcitrant class of acceptors. PMID- 12744703 TI - Molecular dynamics simulations to determine the optimal loop length in the helix loop-helix motif. AB - In this study, 100 ps molecular dynamics simulations were conducted to determine the optimal length of the Gly loop in the helix-loop-helix motif. The results revealed that the length of the loop significantly affected the stability of the two alpha-helices. In addition, the optimal loop length to maintain the highest helicity of the target peptide was found to be the one corresponding to 7 Gly residues. Longer loop length resulted in structural destabilization due to the entropic effect, while shorter loop length was not suitable to achieve intrachain packing of the two alpha-helices. PMID- 12744702 TI - Probing cofactor specificity in phenylalanine hydroxylase by molecular dynamics simulations. AB - Phenylalanine hydroxylase (PAH) is a tetrahydrobiopterin-dependent enzyme that catalyzes the hydroxylation of L-phenylalanine (L-Phe) to L-tyrosine using dioxygen as an additional substrate. The requirement of PAH for a cofactor is absolute, but several cofactor analogs are able to substitute the natural cofactor in catalysis. However, it is only the natural cofactor 6R tetrahydrobiopterin (6R-BH(4)) that induces a negative regulatory effect on the enzyme. In order to get further insights on the molecular basis for this specificity, we studied the structure of the cofactor-enzyme complex and the conformational changes induced by cofactor binding by molecular dynamics simulations. Simulations were carried out on the enzyme alone and complexed with 6R-BH(4) and with two cofactor analogs, 6S-BH(4) and 6-methyl-tetrahydropterin (6M-PH(4)). In the resting unbound enzyme Tyr377 in the catalytic domain is hydrogen bonded to both Ser23 and Glu21 of the autoregulatory N-terminal sequence. This hydrogen bonding network is disturbed by the binding of BH(4), which interacts with Ser23. By doing so, 6R-BH(4) facilitates an interaction between Glu21 and the active site iron, further pulling the N-terminal into the active site of PAH and blocking the L-Phe binding site. Thus, in the 6R-BH(4) complexed enzyme, the N-terminal functions as an intrinsic amino acid regulatory sequence (IARS). Neither 6M-PH(4) nor 6S-BH(4) can interact favorably with Ser23, and do not induce an inhibitory effect on PAH. These simulations thus explain the previous findings that the two hydroxyl groups in the side chain of the 6R epimer of BH(4) are essential for the inhibitory regulatory effect on PAH. PMID- 12744704 TI - Multi-dimensional linguistic complexity. AB - Linguistic complexity is a simple and elegant way of calculating complexity of strings of data. It is based on the concept that the greater the vocabulary one uses, the more complex the data. Until now, it has been used only on one dimensional data, such as DNA and protein sequences and various human language texts. The basic definition can be extended to higher dimensions, thus allowing a practical and simple calculation of linguistic complexity of images, 3D objects and other multi-dimensional data. A simple extension of linguistic complexity is introduced, followed by 2D presentations and a discussion of parametric considerations. An example of linguistic complexity calculations, demonstrating its image processing and medical diagnostic power is presented. The subjects of this paper are patent application pending. PMID- 12744705 TI - Plausible interaction of an alpha-fetoprotein cyclopeptide with the G-protein coupled receptor model GPR30: docking study by molecular dynamics simulated annealing. AB - In this manuscript, the procedure of molecular dynamics simulated annealing is applied to locate a probable receptor and binding site of a cyclicpeptide that inhibits estrogen-stimulated proliferation of breast cancer. The hydrophilic cyclopeptide EMTOVNOGQ (O = 4-hydroxyproline), derived from alpha-fetoprotein, is an inhibitor of estrogen-stimulated proliferation of human breast cancer. This peptide has been shown to act through a mechanism different from that of estrogen; however, its receptor is unknown. We report computer experiments that suggest that this peptide may execute its actions by interacting with GPR30, a G protein-coupled receptor. The subject of this work is the simulation, by molecular dynamics simulated annealing, of the interaction of cyclopeptide EMTOVNOGQ with receptor GPR30 protein. A conformational analysis of the cyclopeptide was undertaken and the final structure was docked on several sites of the GPR30 3D model. Our results show that the cyclopeptide interacts on the pocket located between TM6 and TM7 transmembrane helices of the G-protein, triggering a slight conformational change in the secondary structure of the receptor in the complex. Based on differences in accessible surface areas between GPR30 and its ligand, the residues in the interaction zone were identified. The cyclopeptide is stabilized in the active site by forming a network of hydrogen bonds between Glu, Thr, (1)Pro(OH) and GLn residues of the ligand and Arg-259, Cys-271, Asn-316, Asn-320 and Tyr-324 of the G-protein. Moreover, the study of the electrostatic surface potential on the GPR30 receptor shows that the active site is more positively charged than the other sites. Our modeling indicates a plausible interaction of the cyclopeptide with the seven transmembrane GPR30 protein. This may have profound implications for the treatment of breast cancer. PMID- 12744707 TI - Conformational characteristics and correlations in crystal structures of nucleic acid oligonucleotides: evidence for sub-states. AB - Sugar phosphate backbone conformations are a structural element inextricably involved in a complete understanding of specific recognition nucleic acid ligand interactions, from early stage discrimination of the correct target to complexation per se, including any structural adaptation on binding. The collective results of high resolution DNA, RNA and protein/DNA crystal structures provide an opportunity for an improved and enhanced statistical analysis of standard and unusual sugar-phosphate backbone conformations together with corresponding dinucleotide sequence effects as a basis for further exploration of conformational effects on binding. In this study, we have analyzed the conformations of all relevant crystal structures in the nucleic acids data base, determined the frequency distribution of all possible epsilon, zeta, alpha, beta and gamma backbone angle arrangements within four nucleic acid categories (A-RNA and A-DNA, free and bound B-DNA) and explored the relationships between backbone angles, sugar puckers and selected helical parameters. The trends in the correlations are found to be similar regardless of the nucleic acid category. It is interesting that specific structural effects exhibited by the different unusual backbone sub-states are in some cases contravariant. Certain alpha/gamma changes are accompanied by C3' endo (north) sugars, small twist angles and positive values of base pair roll, and favor a displacement of nucleotide bases towards the minor groove compared to that of canonical B form structures. Unusual epsilon/zeta combinations occur with C2' (south) sugars, high twist angles, negative values of base pair roll, and base displacements towards the major groove. Furthermore, any unusual backbone correlates with a reduced dispersion of equilibrium structural parameters of the whole double helix, as evidenced by the reduced standard deviations of almost all conformational parameters. Finally, a strong sequence effect is displayed in the free oligomers, but reduced somewhat in the ligand bound forms. The most variable steps are GpA and CpA, and, to a lesser extent, their partners TpC and TpG. The results provide a basis for considering if the variable and non-variable steps within a biological active sequence precisely determine morphological structural features as the curvature direction, the groove depth, and the accessibility of base pair for non covalent associations. PMID- 12744706 TI - Coordination and thermodynamics of stable Zn(II) complexes in the gas phase. AB - Ab initio molecular orbital methods in combination with DFT calculations were used to study the structural and thermodynamic properties of 17 complexes containing zinc cation and four first-shell ligands as models of active site of metalloenzymes (e.g. angiotensin converting enzyme, thermolysin). The geometry of the complexes was relaxed by complete optimization by ab initio molecular orbital methods at Hertree-Fock level with 3-21G* basis set. Following single point calculation with tight SCF criteria at the B3LYP level with 6-311+G(2d,p) basis set was used to calculate accurate interaction enthalpies. The structure and thermodynamics of optimized complexes are discussed from the point of view of their biological importance. PMID- 12744708 TI - Ethidium probing of the parallel double- and four-stranded structures formed by the telomeric DNA sequences dG(GT)4G and d(GT)5. AB - Oligonucleotides 3'-d(GT)(5)-(CH(2)CH(2)O)(3)-d(GT)(5)-3' (parGT), containing GT repeats present in the telomeric DNA from Saccharomyces cerevisiae, had been demonstrated to form bimolecular structure, GT-quadruplex (qGT) [O. F. Borisova et al. FEBS Letters 306, 140-142 (1992)]. Four d(GT)(5) strands of the GT quadruplex are parallel and form five G-quartets while thymines are bulged out. The four GT repeats when flanked by guanines, 3'-dG(TG)(4)G-(CH(2)CH(2)O)(3) dG(GT)(4)G-3' (hp-GT), had been shown to form a novel parallel-stranded (ps) double helix with G.G and T.T base pairs (hp-GT ps-DNA) [A. K. Shchyolkina et al. J. Biomol. Struct. Dyn. 18, 493-503 (2001)]. In the present study the intercalator ethidium bromide (Et) was used for probing the two structures. The mode of Et binding and its effect on thermostability of qGT and hp-GT were compared. The quantum yield (q) and the fluorescence lifetime (tau) of Et:qGT (q = 0.15 +/- 0.01 and tau = 24 +/- 1 ns) and Et:hp-GT (q = 0.10 +/- 0.01 and tau = 16.5 +/- 1 ns) indicative of intercalation mode of Et binding were determined. Et binding to qGT was found to be cooperative with corresponding coefficient omega = 3.9 +/- 0.1 and the binding constant Kappa = (6.4 +/- 0.1).10(4) M(-1). The maximum number of Et molecules intercalating into GT-quadruplex is as high as twice the number of innerspaces between G-quartets (eight in our case). The data conform to the model of Et association with GT-quadruplex suggested earlier [O. F. Borisova et al. Mol. Biol. (Russ) 35, 732-739 (2001)]. The anticooperative type of Et binding was observed in case of hp-GT ps-DNA, with the maximum number of bound Et molecules, N = 4 / 5, and the association constant Kappa = (1.5 +/- 0.1).10(5) M(-1). Thermodynamic parameters of formation of Et:qGT and EtBr:hp-GT complexes were calculated from UV thermal denaturation profiles. PMID- 12744709 TI - Na2CO3 influence on DNA double helix stability: strong anion destabilizing effect. AB - Addition of Na(2)CO(3) to almost salt-free DNA solution (5.10(-5)M EDTA, pH=5.7, T(m)=26.5 degrees C) elevates both pH and the DNA melting temperature (T(m)) if Na(2)CO(3) concentration is less than 0.004 M. For 0.004 M Na(2)CO(3), T(m)=58 degrees C is maximal and pH=10.56. Further increase in concentration gives rise to a monotonous decrease in T(m) to 37 degrees C for 1M Na(2)CO(3) (pH=10.57). Increase in pH is also not monotonous. The highest pH=10.87 is reached at 0.04 M Na(2)CO(3) (T(m)=48.3 degrees C). To reveal the cause of this DNA destabilization, which happens in a narrow pH interval (10.56/10.87) and a wide Na(2)CO(3) concentration interval (0.004/1M), a procedure has been developed for determining the separate influences on T(m) of Na(+), pH, and anions formed by Na(2)CO(3) (HCO(3)(-) and CO(3)(2-)). Comparison of influence of anions formed by Na(2)CO(3) on DNA stability with Cl(-) (anion inert to DNA stability), ClO(4)(-) (strong DNA destabilizing "chaotropic" anion) and OH(-) has been carried out. It has been shown that only Na(+) and pH influence T(m) in Na(2)CO(3) solution at concentrations lower than 0.001 M. However, the T(m) decrease with concentration for [Na(2)CO(3)]>/=0.004 M is only partly caused by high pH=10.7. Na(2)CO(3) anions also exert a strong destabilizing influence at these concentrations. For 0.1M Na(2)CO(3) (pH=10.84, [Na(+)]=0.2M, T(m)=42.7 degrees C), the anion destabilizing effect is higher 20 degrees C. For NaClO(4) (ClO(4)(-) is a strong "chaotropic" anion), an equal anion effect occurs at much higher concentrations approximately 3M. This means that Na(2)CO(3) gives rise to a much stronger anion effect than other salts. The effect is pH dependent. It decreases fivefold at neutral pH after addition of HCl to 0.1M Na(2)CO(3) as well as after addition of NaOH for pH greater than 11.2. PMID- 12744710 TI - Theoretical ab initio study of the effects of methylation on structure and stability of G:C Watson-Crick base pair. AB - Methylation of DNA occurs most readily at N(3), N(7), and O(6) of purine bases and N(3) and O(2) of pyrimidines. Methylated bases are continuously formed through endogenous and exogenous mechanisms. The results of a theoretical ab initio study on the methylation of G:C base pair components are reported. The geometries of the local minima were optimized without symmetry restrictions by the gradient procedure at DFT level of theory and were verified by energy second derivative calculations. The standard 6-31G(d) basis set was used. The single point calculations have been performed at the MP2/6-31G(d,p), MP2/6-31++G(d,p), and MP2/6-311++G(2d,2p) levels of theory. The geometrical parameters, relative stability and counterpoise corrected interaction energies are reported. Also, using a variation-perturbation energy decomposition scheme we have found the vital contributions to the total interaction energy. PMID- 12744711 TI - Molecular structure and properties of protonated and methylated derivatives of cytosine. AB - Ab initio techniques were used to determine the effects of protonation and methylation on cytosine's molecular geometry, molecular affinities, relative stability, and structural rigidity. The geometries of local minima were fully optimized by the gradient procedure at DFT and MP2 levels of theory with the medium size 6-31G(d,p) basis set. The results of energetic analysis indicate that N(3)-methyl-cytosine and C(5)-methyl-cytosine are the most stable derivatives for monocationic and neutral species, respectively. The structural rigidity of each species was assessed by an analysis of normal out-of-plane frequencies, the amplitudes, and by the contribution of internal coordinates to the potential energy distributions. The obtained evidence suggests that methylation increases the overall structural flexibility of cytosine and that all molecules in this study populate a non-planar conformation 50% of the time. PMID- 12744713 TI - Marijuana as doping in sports. AB - A high incidence of positive cases for cannabinoids, in analyses for doping control in sports, has been observed since the International Olympic Committee (IOC) included them in the 1989 list of prohibited drugs under the title of classes of prohibited substances in certain circumstances. Where the rules of sports federations so provide, tests are conducted for marijuana, hashish or any other cannabis product exposure by means of urinalysis of 11-nor-delta-9 tetrahydrocannabinol-9-carboxylic acid (carboxy-THC) the main metabolite of delta 9-tetrahydrocannabinol (THC). Concentrations >15 ng/mL (cut-off value) in confirmatory analytical procedures are considered doping. Cannabis is an illicit drug in several countries and has received much attention in the media for its potential therapeutic uses and the efforts to legalise its use. Studies have demonstrated that the use of cannabinoids can reduce anxiety, but it does not have ergogenic potential in sports activities. An increase in heart rate and blood pressure, decline of cardiac output and reduced psychomotor activity are some of the pharmacological effects of THC that will determine a decrease in athletic performance. An ergolytic activity of cannabis products has been observed in athletes of several different sport categories. In Brazil, analyses for doping control in sports, performed in our laboratories, have detected positive cases for carboxy-THC in urine samples of soccer, volleyball, cycling and other athletes. It is our intention to discuss in this article some points that may discourage individuals from using cannabis products during sports activities, even in the so-called permitted circumstances defined by the IOC and some sports federations. PMID- 12744714 TI - Allergic rhinoconjunctivitis in elite athletes: optimal management for quality of life and performance. AB - Allergic rhinoconjunctivitis is a common condition with a peak incidence in the age range of the majority of elite athletes. The condition has been shown to have a significant impact on the quality of life of those affected and poses particular challenges when present in the elite athlete. When an athlete is looking for exceptional performance at events such as the Olympic Games, any factor which affects quality of life by interfering with sleep, decreasing the ability to concentrate, or reducing peak physical fitness, may have a significant impact on the ability to perform at one's best. Optimal management begins with correct diagnosis and identification of triggering factors. There are a number of therapeutic options available to the treating physician. When formulating a management plan for the elite athlete, the physician must consider "doping" rules and the possible effect of medication on athletic performance. Medication choices include the newer, non-sedating antihistamines, used either orally or topically, and the prophylactic use of intranasal corticosteroids. When allergic conjunctivitis is the principal problem, the newer, topical antihistamines are highly effective and have a rapid onset of action. Since avoidance strategies are rarely practical for the athlete, consideration should be given to strategies such as immunotherapy, where long-term benefit is possible. PMID- 12744716 TI - Movement velocity in resistance training. AB - Guidelines for resistance training include the number of exercises, sets, repetitions, and frequency of training, but only vaguely mention movement velocity. Nevertheless, different velocities imply different performances, i.e. a different number of repetitions or different loads. Studies investigating the effect of different movement velocities on resistance training have not reached a consensus. Some studies indicate specificity in strength gains while others indicate generality. Some indicate slow training to be better, others indicate fast training, and still others indicate no differences. Most of these studies were conducted on isokinetic or hydraulic equipment. Very few used isotonic equipment for training, and their results seem to tend towards generality and no differences in gains of strength between velocities. Defining the training velocity is mostly important for athletic performances where a wide range of velocities is needed and transfer of gains would greatly optimise training. At the other end of the spectrum, there are the most frail and elderly, to whom power loss may impair even daily functions, but training with fast velocities might increase injury risk and, therefore, transfer of gains from slow training would be greatly beneficial. Movement velocity for resistance training with isotonic equipment needs to be further investigated so that recommendations may be made. PMID- 12744715 TI - The concept of maximal lactate steady state: a bridge between biochemistry, physiology and sport science. AB - The maximal lactate steady state (MLSS) is defined as the highest blood lactate concentration (MLSSc) and work load (MLSSw) that can be maintained over time without a continual blood lactate accumulation. A close relationship between endurance sport performance and MLSSw has been reported and the average velocity over a marathon is just below MLSSw. This work rate delineates the low- to high intensity exercises at which carbohydrates contribute more than 50% of the total energy need and at which the fuel mix switches (crosses over) from predominantly fat to predominantly carbohydrate. The rate of metabolic adenosine triphosphate (ATP) turnover increases as a direct function of metabolic power output and the blood lactate at MLSS represents the highest point in the equilibrium between lactate appearance and disappearance both being equal to the lactate turnover. However, MLSSc has been reported to demonstrate a great variability between individuals (from 2-8 mmol/L) in capillary blood and not to be related to MLSSw. The fate of enhanced lactate clearance in trained individuals has been attributed primarily to oxidation in active muscle and gluconeogenesis in liver. The transport of lactate into and out of the cells is facilitated by monocarboxylate transporters (MCTs) which are transmembrane proteins and which are significantly improved by training. Endurance training increases the expression of MCT1 with intervariable effects on MCT4. The relationship between the concentration of the two MCTs and the performance parameters (i.e. the maximal distance run in 20 minutes) in elite athletes has not yet been reported. However, lactate exchange and removal indirectly estimated with velocity constants of the individual blood lactate recovery has been reported to be related to time to exhaustion at maximal oxygen uptake. PMID- 12744717 TI - Warm up I: potential mechanisms and the effects of passive warm up on exercise performance. AB - Despite limited scientific evidence supporting their effectiveness, warm-up routines prior to exercise are a well-accepted practice. The majority of the effects of warm up have been attributed to temperature-related mechanisms (e.g. decreased stiffness, increased nerve-conduction rate, altered force-velocity relationship, increased anaerobic energy provision and increased thermoregulatory strain), although non-temperature-related mechanisms have also been proposed (e.g. effects of acidaemia, elevation of baseline oxygen consumption (.VO(2)) and increased postactivation potentiation). It has also been hypothesised that warm up may have a number of psychological effects (e.g. increased preparedness). Warm up techniques can be broadly classified into two major categories: passive warm up or active warm up. Passive warm up involves raising muscle or core temperature by some external means, while active warm up utilises exercise. Passive heating allows one to obtain the increase in muscle or core temperature achieved by active warm up without depleting energy substrates. Passive warm up, although not practical for most athletes, also allows one to test the hypothesis that many of the performance changes associated with active warm up can be largely attributed to temperature-related mechanisms. PMID- 12744718 TI - Anterior cruciate ligament injury in paediatric and adolescent patients: a review of basic science and clinical research. AB - Anterior cruciate ligament (ACL) injuries are recognised with greater frequency in children and adolescents. Non-operative treatment of ACL injuries in children may lead to knee instability and secondary injuries, especially in those who return to sports. ACL reconstruction is controversial in skeletally immature patients because of potential damage to the proximal tibial and distal femoral physes, which may lead to premature arrest and/or leg length discrepancies. This paper reviews studies of ACL injuries in children and adolescents, and examines basic science and clinical studies concerning physeal arrest secondary to ACL reconstruction tunnels. Some animal studies support the conclusion that ACL reconstructions in children have the potential to cause growth disturbances, and there are reports of growth plate complications due to ACL reconstruction in skeletally immature patients. There is evidence that ACL reconstruction can be performed in select skeletally immature patients, but the risk of growth plate complications must be considered. PMID- 12744719 TI - Rab5 and Rab11 mediate transferrin and anti-variant surface glycoprotein antibody recycling in Trypanosoma brucei. AB - The mammalian-infective bloodstream form of Trypanosoma brucei possesses a highly active endocytotic system. Evasion of the host immune response by T. brucei is dependent on antigenic variation of VSG (variant surface glycoprotein), but additional mechanisms for removal of surface-bound antibody also operate. Four Rab proteins, Tb (trypanosomal) RAB4, 5A, 5B and 11 are located to the endosomal system; TbRAB5A and TbRAB11 co-localize with internalized anti-VSG antibody and transferrin. A live cell assay was used to record a single cycle of endocytosis of anti-VSG IgG and transferrin, their subsequent degradation within the endosomal system and exocytosis of the products. TbRAB5A and TbRAB11 were involved in the overall process of endocytosis, degradation and exocytosis, whereas TbRAB5B and TbRAB4 were not implicated. The kinetics of anti-VSG IgG and transferrin recycling depend on the nucleotide state of TbRAB5A and TbRAB11. These data, together with previous work, suggest that IgG and transferrin initially enter a TbRAB5A sorting endosome and are most probably recycled subsequently via a TbRAB11-dependent step. Analysis of the recycled IgG and transferrin demonstrated extensive degradation of these recycled proteins. Degradation of transferrin was enhanced in cells expressing increased amounts of TbRAB5A or TbRAB11 with a Ser-->Asn mutation, but was decreased when active TbRAB11 was overexpressed. The extent of degradation of anti-VSG IgG was found to be unaffected by mutant Rab protein expression. The presence of an efficient mechanism for the removal of IgG bound to the external surface of T. brucei and its subsequent proteolysis within the recycling system suggests a role for this pathway in immune evasion. PMID- 12744720 TI - Mouse matriptase-2: identification, characterization and comparative mRNA expression analysis with mouse hepsin in adult and embryonic tissues. AB - We report the identification and characterization of mouse matriptase-2 (m matriptase-2), an 811-amino-acid protein composed of an N-terminal cytoplasmic domain, a membrane-spanning domain, two CUB (complement protein subcomponents C1r/C1s, urchin embryonic growth factor and bone morphogenetic protein 1) domains, three LDLR (low-density-lipoprotein receptor class A) domains and a C terminal serine-protease domain. All m-matriptase-2 protein domain boundaries corresponded with intron/exon junctions of the encoding gene, which spans approx. 29 kb and comprises 18 exons. Matriptase-2 is highly conserved in human, mouse and rat, with the rat matriptase-2 gene ( r-maltriptase-2 ) predicted to encode transmembrane and soluble isoforms. Western-blot analysis indicated that m matriptase-2 migrates close to its theoretical molecular mass of 91 kDa, and immunofluorescence analysis was consistent with the proposed surface membrane localization of this protein. Reverse-transcription PCR and in-situ hybridization analysis indicated that m-matriptase-2 expression overlaps with the distribution of mouse hepsin (m-hepsin, a cell-surface serine protease identified in hepatoma cells) in adult tissues and during embryonic development. In adult tissues both are expressed at highest levels in liver, kidney and uterus. During embryogenesis m-matriptase-2 expression peaked between days 12.5 and 15.5. m hepsin expression was biphasic, with peaks at day 7.5 to 8.5 and again between days 12.5 and 15.5. In situ hybridization of embryonic tissues indicated abundant expression of both m-matriptase-2 and m-hepsin in the developing liver and at lower levels in developing pharyngo-tympanic tubes. While m-hepsin was detected in the residual embryonic yolk sac and with lower intensity in lung, heart, gastrointestinal tract, developing kidney tubules and epithelium of the oral cavity, m-matriptase-2 was absent in these tissues, but strongly expressed within the nasal cavity by olfactory epithelial cells. Mechanistic insight into the potential role of this new transmembrane serine protease is provided by its novel expression profile in embryonic and adult mouse. PMID- 12744722 TI - Sedation in palliative care - a critical analysis of 7 years experience. AB - BACKGROUND: The administration of sedatives in terminally ill patients becomes an increasingly feasible medical option in end-of-life care. However, sedation for intractable distress has raised considerable medical and ethical concerns. In our study we provide a critical analysis of seven years experience with the application of sedation in the final phase of life in our palliative care unit. METHODS: Medical records of 548 patients, who died in the Palliative Care Unit of GK Havelhoehe between 1995-2002, were retrospectively analysed with regard to sedation in the last 48 hrs of life. The parameters of investigation included indication, choice and kind of sedation, prevalence of intolerable symptoms, patients' requests for sedation, state of consciousness and communication abilities during sedation. Critical evaluation included a comparison of the period between 1995-1999 and 2000-2002. RESULTS: 14.6% (n = 80) of the patients in palliative care had sedation given by the intravenous route in the last 48 hrs of their life according to internal guidelines. The annual frequency to apply sedation increased continuously from 7% in 1995 to 19% in 2002. Main indications shifted from refractory control of physical symptoms (dyspnoea, gastrointestinal, pain, bleeding and agitated delirium) to more psychological distress (panic stricken fear, severe depression, refractory insomnia and other forms of affective decompensation). Patients' and relatives' requests for sedation in the final phase were significantly more frequent during the period 2000-2002. CONCLUSION: Sedation in the terminal or final phase of life plays an increasing role in the management of intractable physical and psychological distress. Ethical concerns are raised by patients' requests and needs on the one hand, and the physicians' self-understanding on the other hand. Hence, ethically acceptable criteria and guidelines for the decision making are needed with special regard to the nature of refractory and intolerable symptoms, patients' informed consent and personal needs, the goals and aims of medical sedation in end-of-life care. PMID- 12744721 TI - N-linked oligosaccharides play a role in disulphide-dependent dimerization of intestinal mucin Muc2. AB - Within the C-terminal domain of many secretory mucins is a 'cystine knot' (CK), which is needed for dimer formation in the endoplasmic reticulum. Previous studies indicate that in addition to an unpaired cysteine, the three intramolecular cystine bonds of the knot are important for stability of the dimers formed by rat intestinal mucin Muc2. The present study was undertaken to determine whether the two N-glycans N9 and N10, located near the first and second cysteines of the knot, also play a role in dimer formation. The C-terminal domain of rat Muc2 (RMC), a truncated RMC mutant containing the CK, and mutants lacking N9 and N10 sites, were expressed in COS-1 cells and the products monitored by radioactive [(35)S]Met/Cys metabolic pulse-chase and immunoprecipitation. Mutation of N9, but not N10, caused increased synthesis of dimers over a 2-h chase period. The N9 mutant remained associated with calreticulin for a prolonged period. About 34-38% of the total labelled products of RMC and its mutants was secreted into the media by 2 h, but the proportion in dimer form was dramatically reduced for the N9 mutant, suggesting lower dimer stability relative to RMC or its N10 mutant. We conclude that under normal conditions the presence of the N9 glycan functions to maintain a folding rate for mucin monomers that is sufficiently slow to allow structural maturation and stability of Muc2 dimers. To our knowledge this report is the first demonstration that a specific N-glycan plays a definitive role in mucin dimer formation. PMID- 12744723 TI - Chemical cleavage reactions of DNA on solid support: application in mutation detection. AB - BACKGROUND: The conventional solution-phase Chemical Cleavage of Mismatch (CCM) method is time-consuming, as the protocol requires purification of DNA after each reaction step. This paper describes a new version of CCM to overcome this problem by immobilizing DNA on silica solid supports. RESULTS: DNA test samples were loaded on to silica beads and the DNA bound to the solid supports underwent chemical modification reactions with KMnO4 (potassium permanganate) and hydroxylamine in 3M TEAC (tetraethylammonium chloride) solution. The resulting modified DNA was then simultaneously cleaved by piperidine and removed from the solid supports to afford DNA fragments without the requirement of DNA purification between reaction steps. CONCLUSIONS: The new solid-phase version of CCM is a fast, cost-effective and sensitive method for detection of mismatches and mutations. PMID- 12744724 TI - Measurement of illumination exposure in postpartum women. AB - BACKGROUND: Low levels of light exposure at critical times are thought to cause seasonal affective disorder. Investigators, in studies demonstrating the usefulness of bright light therapy, also have implicated light's role in non seasonal depression. The precise cause of postpartum depression has not been delineated, but it seemed possible that new mothers would spend reduced time in daylight. The goal of this study was to examine the levels of illumination experienced by postpartum mothers and to discover any relationship between light exposure and mood levels experienced during the postpartum period. METHODS: Fifteen postpartum women, who did not have any baseline indication of depression, wore a wrist device (Actillume) for 72 hours to measure their exposure to light. At the end of the recording period, they completed a self-reported measure of mood. The mean light exposure of these postpartum women (expressed as the 24-hour average logarithm of illumination in lux) was compared with that of a representative sample of women of comparable age, residence, and seasonal months of recording. Mood levels were then rank-ordered and tested for correlation with light exposure levels. RESULTS: There was no significant difference between the amount of light [log10lux] experienced by postpartum (1.01 SD 0.236) and control women (1.06 SD 0.285). Mood was not correlated with illumination in the postpartum sample. CONCLUSIONS: Postpartum women in San Diego did not receive reduced light, nor was low mood related to low illumination. PMID- 12744727 TI - Shape differences rather than size differences between castes in the Neotropical swarm-founding wasp Metapolybia docilis (Hymenoptera: Vespidae, Epiponini). AB - BACKGROUND: Swarm-founding epiponine wasps are an intriguing group of social insects in which colonies are polygynic (several queens share reproduction) and differentiation between castes is often not obvious. However, caste differences in some may be more pronounced in later phases of the colony cycle. RESULTS: Using morphometric analyses and multivariate statistics, it was found that caste differences in Metapolybia docilis are slight but more distinct in latter stages of the colony cycle. CONCLUSIONS: Because differences in body parts are so slight, it is proposed that such variation may be due to differential growth rates of body parts rather than to queens being larger in size, similar to other previously observed epiponines. PMID- 12744725 TI - Biological-effective versus conventional dose volume histograms correlated with late genitourinary and gastrointestinal toxicity after external beam radiotherapy for prostate cancer: a matched pair analysis. AB - BACKGROUND: To determine whether the dose-volume histograms (DVH's) for the rectum and bladder constructed using biological-effective dose (BED-DVH's) better correlate with late gastrointestinal (GI) and genitourinary (GU) toxicity after treatment with external beam radiotherapy for prostate cancer than conventional DVH's (C-DVH's). METHODS: The charts of 190 patients treated with external beam radiotherapy with a minimum follow-up of 2 years were reviewed. Six patients (3.2%) were found to have RTOG grade 3 GI toxicity, and similarly 6 patients (3.2%) were found to have RTOG grade 3 GU toxicity. Average late C-DVH's and BED DVH's of the bladder and rectum were computed for these patients as well as for matched-pair control patients. For each matched pair the following measures of normalized difference in the DVH's were computed: (a) deltaAUC = (Area Under Curve [AUC] in grade 3 patient--AUC in grade 0 patient)/(AUC in grade 0 patient) and (b) deltaV60 = (Percent volume receiving = 60 Gy [V60] in grade 3 patient- V60 in grade 0 patient)/(V60 in grade 0 patient). RESULTS: As expected, the grade 3 curve is to the right of and above the grade 0 curve for all four sets of average DVH's--suggesting that both the C-DVH and the BED-DVH can be used for predicting late toxicity. deltaAUC was higher for the BED-DVH's than for the C DVH's--0.27 vs 0.23 (p = 0.036) for the rectum and 0.24 vs 0.20 (p = 0.065) for the bladder. deltaV60 was also higher for the BED-DVH's than for the C-DVH's- 2.73 vs 1.49 for the rectum (p = 0.021) and 1.64 vs 0.71 (p = 0.021) for the bladder. CONCLUSIONS: When considering well-established dosimetric endpoints used in evaluating treatment plans, BED-DVH's for the rectum and bladder correlate better with late toxicity than C-DVH's and should be considered when attempting to minimize late GI and GU toxicity after external beam radiotherapy for prostate cancer. PMID- 12744728 TI - [Bone marrow endothelial cell-conditioned medium promotes hematopoietic differentiation of mouse embryonic stem cells]. AB - The purpose of this study was to observe the bone marrow endothelial cell conditioned medium (BECM) and cytokines, i.e. vascular endothelial growth factor (VEGF), stem cell factor (SCF) and EPO promoting the generation of hematopoietic precursor cells from mouse embryonic stem cells (ESC) in vitro. Day 4 embryoid body (4dEB) cells were derived from ESC-D3 cell line, a murine ESC line, and then induced with BECM and/or cytokines. Four groups, i.e. BECM, BECM + VEGF + SCF + EPO, VEGF + SCF + EPO and control (spontaneous differentiation), were designed. Immunochemistry staining and flow cytometry were adopted to observe the antigen expression, RT-PCR to detect hematopoietic transcription factors, and hematopoietic progenitor assay to examine hematopoietic differentiation. The results showed that the cells induced from ESC expressed hematopoietic precursor cell antigens (c-kit, Sca-1, Thy-1 and CD34), transcription factors (c-myb, SCL and beta-H1) and generated HPP-CFC and BFU-E. The effect of BECM + VEGF + SCF + EPO was the most potent in the inducing groups according to the numbers of hematopoietic precursor cells and colonies. It is concluded that BECM promotes the differentiation of ESC into hematopoietic precursor cells in vitro, and this effect is the strongest when BECM combining with VEGF + SCF + EPO. PMID- 12744729 TI - [Human bone marrow mesenchymal stem cells express multiple hematopoietic growth factors]. AB - To study the biological role of human cultured bone marrow mesenchymal stem cell (BM-MSC) in hematopoiesis by investigation of its expression of multiple hematopoietic growth factors, RT-PCR was used to analyze the expression of SCF, Flt3-ligand, TPO, LIF, G-CSF, GM-CSF, IL-3, IL-6 and IL-11 at mRNA level for human BM-MSC from healthy donors and patients with leukemia and lymphoma. BM-MSC were incubated with or without hydrocortison (HC). The results clearly showed that the cultured BM-MSC expressed mRNA of SCF, Flt3-ligand, TPO, LIF, IL-6 and IL-11 at passages 3 up to 15, but did not express G-CSF, GM-CSF and IL-3. The same expression pattern of above cytokines was seen also for the patient's BM MSC. HC was able to induce BM-MSC to express G-CSF but not to express GM-CSF. BM MSC seemed not to change morphologically after incubation with HC for up to 21 days. In conclusion, both normal and patient BM-MSC should be potential to promote hematopoiesis according to their expression of multiple hematopoietic cytokines, and HC is able to induce hematopoietic growth factor expression. PMID- 12744726 TI - A high resolution physical and RH map of pig chromosome 6q1.2 and comparative analysis with human chromosome 19q13.1. AB - BACKGROUND: The generation of BAC/PAC contigs in targeted genome regions is a powerful method to establish high-resolution physical maps. In domestic animal species the generation of such contigs is typically initiated with the screening of libraries with probes derived from human genes that are expected to be located in the region of interest by comparative mapping. However, in many instances the available gene-derived probes are too far apart to allow the cloning of BAC/PAC contigs larger than a few hundred kb. High resolution physical mapping allows to estimate the sizes of gaps and to control the orientation of the individual sub contigs, which helps to avoid errors during the assembly of smaller contigs into final Mb-sized contigs. The recently constructed porcine IMNpRH2 panel allowed us to use this approach for the construction of high-resolution physical maps of SSC 6q1.2. RESULTS: Two sequence-ready BAC/PAC contigs of the gene-rich region on porcine chromosome 6q1.2 (SSC 6q1.2) containing the RYRl gene were constructed. The two contigs spanned about 1.2 Mb and 2.0 Mb respectively. The construction of these contigs was monitored by the results provided by the mapping of 15 markers on the IMpRH(7000rad) and 35 markers on the IMNpRH2(12000rad) radiation hybrid panels. Analyses on the IMpRH panel allowed us to globally link and orientate preliminary smaller contigs, whereas analyses on the high resolution IMNpRH2 panel allowed us to finally identify the order of genes and markers. CONCLUSIONS: A framework map of 523 cR12000 was established covering the whole studied region. The order of markers on the framework 1000:1 RH map was found totally consistent with the data deduced from the contig map. The kb/cR ratio was very constant in the whole region, with an average value of 6.6 kb/cR. We estimate that the size of the remaining gap between the two contigs is of about 300 kb. The integrated physical and RH map of the investigated region on SSC 6q1.2 was used for a comparative analysis with respect to the syntenic regions on HSA 19q13.1 and MMU 7 and revealed a perfectly conserved gene order across the entire studied interval. PMID- 12744730 TI - [Proliferation and differentiation of human CD34+ hematopoietic stem/progenitor cells induced by Panax notoginosides]. AB - The object of this study was to explore the effects of Panax notoginosides (PNS) on proliferation and differentiation of human CD34(+) stem/progenitor cells. CD34(+) cells were isolated from human bone marrow by using immune beads of Dynal M- 450 system. The cells were exposed to PNS at different concentrations in both liquid and semi-solid culture for 14 days. The cells were marked with monoclonal antibodies and analyzed by flow cytometry after culture. The CFU-Mix colony formation from CD34(+) cells was assayed. The results showed that: (1) The yield of CD34(+) cells after being selected by immune beads were (1.03 +/- 0.74)% out of bone marrow nuclear cells with purity of 86% - 93%. (2) PNS (10 - 25 mg/L) stimulated the proliferation of CD34(+) cells, and raised the colony numbers of CFU-Mix obviously in vitro. PNS 25 mg/L was the optimal concentration to promote proliferation of CD34(+) cells, the increasing rate of CFU-Mix colony was (34.7 +/- 16.0)%. (3) The differentiation of CD34(+) cells was induced by exposure to PNS (25, 50 and 100 mg/L) in liquid culture for 14 days. The percentages of CD33(+) and CD15(+) cells were increased after PNS exposure, which were significantly higher than those of control (P < 0.01), however CD71(+) and G-A(+) cells were no obviously difference after PNS treatment. In conclusion, Panax notoginosides not only promote the proliferation of CD34(+) cells, but also induce the differentiation committed to granulocytes. PMID- 12744731 TI - [Tissue factor expression in human umbilical vien endothelial cells stimulated by TNF-alpha and its molecular mechanism]. AB - The objective of this study was to explore tissue factor (TF) expression induced by TNF-alpha in cultured human umbilical vien endothelial cells (HUVEC) and its molecular mechanism. TF expression on the surface of HUVEC, TF mRNA and nuclear factor kappaB (NF-kappaB) in HUVEC were detected by flow cytometry, RT-PCR and Western blot respectively. The results showed that TNF-alpha could enhance TF expression on the surface of HUVEC, the TF expression increase was highly consistent with the increased synthesis of TF mRNA, and the increase of TF expression was lately appeared for several hours. It was also found activation of NF kappaB at the time TF mRNA increase. In conclusion, NF-kappaB could be activated promptly after HUVEC incubated with TNF-alpha, then it was bound to TF promotor to start the TF transcription, TF mRNA expression was upregulated, that leaded to the increase of TF expression on the HUVEC surface and activated the coagulation cascade. PMID- 12744732 TI - [Synthesis and function analysis of a new thrombopoietin (TPO) mimic peptide]. AB - In order to find a new TPO-mimic peptide with similar activity to TPO while reducing the side effects, a TPO-mimic peptide (P1) screened from a random peptide library was restructured. The new structure of the TPO mimic peptide (P2) was synthesized. After coupling P2 with Dextran 10 and performing intermolecular oxidation, dextran-coupled and dimerized form of P2 were obtained, naming D-P2 and (P2)(2) respectively. The activities of the peptides in vitro were measured with MTT method. The results showed that the EC50 of P2 was 20 nmol/L, 700 times higher than P1. The EC50 of D-P2 and (P2)(2) were 0.35 nmol/L and 0.14 nmol/L, respectively. After administrating to the mouse, the peptides increased the number of platelets in the blood circulation obviously without influence on other blood cells. In conclusion, the TPO-mimic peptides have prospects in treating diseases related with thrombocytopenia. PMID- 12744733 TI - [Feasibility of real-time quantitative PCR in assessing the efficiency of gene transfection in vivo]. AB - To explore feasibility of real-time quantitative PCR in assessing efficiency of gene transfection, clonal PCR was employed to analyze efficiency of retroviral mediated neo gene transfection in primary myoblast, simultaneous real-time PCR were performed for estimation of transfection efficiency; for measuring integrated gene copy number per cell, linear amplification mediated-PCR (LAM-PCR) and retroviral 5'LTR integration analysis also were used. The results showed that: (1) the data from clonal PCR are similar as that from real-time PCR in low efficiency of transfection (< 36%); but in high efficiency of transfection, it is significantly differentiation between clonal PCR and real-time PCR. (2) One copy of transduced gene per cell was observed in retroviral-mediated gene transfection in primary myoblast. It is concluded that real-time PCR can be used to estimate gene transfer vector in vivo, but it is not available for assessing gene transfection in vitro, because high efficiency of transfection could be obtained in most of gene transfection in vitro. PMID- 12744734 TI - [Analysis of immunophenotype of B-lineage acute lymphoblastic leukemia cells by 4 color flow cytometry]. AB - To investigate the characteristics of immunophenotype of B-lineage acute lymphoblastic leukemia (B-ALL) cells in Chinese cases, B-ALL cells from 181 patients were analyzed by 4-color flow cytometry with CD45/SSC gating. The results demonstrated that the antigen expression frequencies were as follows: CD19 (100%), HLA-DR (98.9%), CD38 (88.5%), CD10 (76.8%) and CD34 (76.8%). CD117 and T antigens (CD2 and CD7) were rarely expressed. Childhood group (or= 19 years) had a higher expression rate of myeloid antigens (CD13 and/or CD33). Subtype CD10(+)/CD34(+) took the highest percentage in three age groups. Percentages of CD10(-)/CD34(+) group increased by age. Subtype CD10( )/CD34(+) had the highest myeloid antigen co-expression. Comparing with CD45 positive cases, those with CD45-negative or CD45(+/-) always had higher CD10 expression. Bcr/abl mRNA was evaluated in 43 samples from patients with B-ALL. Myeloid antigen co-expression was not different in bcr/abl(+) and bcr/abl(-) groups, and m-bcr/abl(+) was mainly observed in subtype CD10(+)/CD34(+). In conclusion, typical B-ALL cells expressed CD19(+), HLA-DR(+), and CD117(-). CD34, CD10 and CD45 expression varied in different maturation stages. The immunophenotypic subtype of B-ALL in adolescents was similar to that in the adults. Blasts with CD45(+) were mostly seen in CD10(-) cases. CD13 and/or CD33 co-expression was fewer in children patients. m-bcr/abl(+) was mainly seen in subtype CD10(+)/CD34(+). PMID- 12744735 TI - [Immunophenotypic features of bcr/abl fusion transcript-positive B-lineage acute lymphoblast leukemia]. AB - To investigate the biological features of leukemic cells in bcr/abl fusion transcript-positive B-lineage acute lymphoblastic leukemia (B-ALL), 3- or 4-color flow cytometry with directly conjugated monoclonal antibodies was used to detect the immunophenotype of the cells in 26 patients with bcr/able-positive B-ALL and 32 patients with bcr/abl-negative B-ALL. bcr/abl fusion transcript was detected by RT-PCR. Immunoglobulin heavy chain (IgH) gene rearrangement was detected by PCR. The results showed that all of the B-ALL patients were positive for CD19. There was significant difference in expression of CD34 (96.2% vs 65.6%), CD10 (96.2% vs 71.8%) and CD38 (43.8% vs 95.4%) between bcr/abl-positive and -negative groups. In bcr/abl-positive B-ALL group, the co-expression rates of CD10(+)/CD19(+)/CD34(+), CD10(+)/CD34(+)/HLA-DR(+) and CD10(+)/CD34(+)/CD38(-) were 92.3% (24/26), 73.1% (19/26) and 56.2% (9/16), respectively. In bcr/abl negative group, co-expression of CD10(+)/CD19(+)/CD34(+) and CD10(+)/CD34(+)/HLA DR(+) were 43.8% (14/32) and 37.5% (12/32), respectively, there were significant differences (P < 0.05) between bcr/abl-positive and -negative groups, but none of the cases co-expressed CD10(+)/CD34(+)/CD38(-). The detection rate of monoclonal IgH gene rearrangement (58.8%, 10/17) was lower in bcr/abl-positive group than that (85.7%, 12/14) in bcr/able-negative group. It is concluded that the expression rates of CD34 and CD10 are higher, and CD38 and IgH gene rearrangement are lower in bcr/abl-positive B-ALL cases, CD10(+)/CD34(+)/CD38(-) is a unique feature of immunophenotype, and this phenotype of leukemia cells is closer to that of early B-lineage progenitor cells. PMID- 12744736 TI - Clinical and laboratory studies of expression of cyclin A in leukemia cells. AB - Uncontrolled cell proliferation is the basic feature of cancer. Some of the prime cell cycle regulators are involved directly in tumorigenesis. Cyclin A, one of the G(1)/S cyclin, can cause transformation. The purpose of this research was to investigate whether cyclin A overexpression was involved in leukemogenesis and proliferation of leukemia cells. The expression of cyclin A at S-phase in leukemia cell line HL-60, blast cells of acute leukemia patients, bone marrow cells of outpatients without malignant hematological disease and peripheral blood cells of healthy donors was investigated by simultaneous indirect immunofluorescence staining of intracellular antigen and DNA. To further investigate whether cyclin A played as a key molecular in cell proliferation, HL 60 cells were exposed to different concentrations of hexamethylene bisacetamide (HMBA). MTT dye absorbance of living cells and cell cycle analysis were adopted to evaluate growth arrest. Differentiation was evaluated by detection of the change of expression of CD11b and CD33 on cell surface. The results showed that overexpression of cyclin A was only found among specimens from acute leukemia and leukemia cell line. There was no elevated cyclin A detection for cyclin A among specimens from outpatients and healthy donors. In HMBA interference experiment, HMBA was able to induce growth arrest and monocytic macrophage differentiation of HL-60 cells in a dose-dependent manner, and all these changes were associated with a marked down-regulation of cyclin A expression. In conclusion, aberrant overexpression of cyclin A at S-phase was only found in leukemia cell lines and blast cells from acute leukemia. The dose-dependent effect of HMBA on cell growth and differentiation of HL-60 cell line which was consistent with the decrease of cyclin A expression in these cells suggested that the molecular mechanisms of HMBA inducement involved downregulation of cyclin A expression. PMID- 12744737 TI - Aberrant expression and deletion of FHIT gene in leukemias. AB - FHIT (fragile histidine triad) gene at chromosome 3p14.2 usually expresses at a very low level in human tissue and cells. A high frequency of abnormalities in FHIT gene has been demonstrated in various cancers. FHIT is proposed as a putative tumor-suppressor gene. To evaluate the expression of the FHIT gene in various leukemias, bone marrow or peripheral blood samples from 98 leukemia patients were tested by RT-PCR: 38 from patients with AML-[M(2)(9), M(3)(12), M(4)(8), M(5)(9)], 16 with ALL, and 34 with CML-[CP(20), AP(4), BC(10)] of various FAB types, as well as 10 patients with other hematological malignancies. To detect a deletion in sequencing the FHIT gene, the representative aberrant PCR products were cloned and then sequenced. The results showed that 22/38 (58%) patients with AML, 9/16 (56%) patients with ALL and 19/34 (56%) patients with CML were detectable of aberrant FHIT mRNA transcripts or deletion of FHIT. In 6 (16%) AML patients, 3 (19%) ALL patients, and 5 (15%) CML patients, the wild-type product was absent. Some patient's samples - 6 (42%) AML, 6 (38%) ALL, and 14 (41%) CML revealed aberrant FHIT transcripts in addition to a normal-sized band. Samples from healthy donors (PB, n = 12; BM, n = 5) did not indicate any abnormal expression. Eleven isolated fragments from various patterns of FHIT gene expression were investigated using cDNA sequencing. Sequence analysis revealed deletion of exon 4-8, exon 5-8, and exon 5-6 in various leukemias, as well as the deletion of the full FHIT gene sequence. The fused transcripts included: exon 3 and exon 9, exon 3 and exon 7, exon 4 and exon 9, exon 5 and exon 7. Sequence analysis of aberrant fragments present in samples from an AML and a CML patients was detected for point mutations and insert mutations located in exons 2, 8 and 10, plus a variety of aberrant transcripts. Deletion or aberrant FHIT mRNA transcripts in 50/98 (51%) leukemia patients were found. All samples with aberrant FHIT lacked gene product. A Kaplan-Meier plot of survival in patients with AML in relation to FHIT expression revealed that aberrance or loss of FHIT gene significantly correlated with a low clinical remission rate and poor overall survival. PMID- 12744738 TI - [Effect of homoharringtonine on expression of human telomerase reverse transcriptase mRNA in HL-60 cells]. AB - To investigate the mechanism and its significance of the depression of telomerase activity in HL-60 cells exposed to homoharringtonine (HHT), the semi-quantitative RT-PCR and PCR-ELISA were used to detect the expression of hTERT mRNA and telomerase activity in HL-60 cells, the apoptotic rate of HL-60 cells was assayed with TUNEL. The results indicated that compared with the control, the level of hTERT mRNA synthesis in HL-60 cells treated with HHT at 0.005 - 0.03 micro g/ml for 12 hours did not change significantly, but it reduced obviously at 0.04 micro g/ml and was undetectable at 0.05 micro g/ml. When HL-60 cells incubated with HHT at 0.02 micro g/ml for various hours, the expression of hTERT mRNA did not decrease after 6 - 18 hours and reduced significantly after 24 hours and was undetectable at 30 hours. The tendency for suppression of telomerase activity was consistent with decrease of the expression of hTERT mRNA in HHT-treated HL-60 cells. The apoptotic rate of HL-60 cells was apparently increased with the depression of hTERT mRNA and telomerase activity. In conclusion, the transcription of hTERT mRNA in HL-60 cells can be inhibited by HHT. The relationship between regulation of hTERT mRNA expression and HHT-induced apoptosis is worth investigating furtherly. PMID- 12744739 TI - [Detection of 20q(-) chromosome abnormality in myelodysplastic syndrome by interphase fluorescence in situ hybridization]. AB - In order to explore the value of interphase fluorescence in situ hybridization (FISH) in the detection of partial deletion of the long arm of chromosome 20 (20q(-)) in patients with myelodysplastic syndrome (MDS), spectrum Green fluorescein directly labeled yeast artificial chromosome (YAC) clone 912C3 which spans the breakpoint cluster region in band 20q12 was used as probes to perform interphase FISH on the marrow cells from 52 cases of MDS and 5 normal controls. 200 to 300 cells were scored for each case and cases which had cells with a green hybridization signal>7.16% were defined as 20q(-) positive. The results of FISH were compared with those of conventional cytogenetics (CC) assay. The results showed that among 52 cases of MDS, 7 (13.5%) cases were positive by FISH, however, of which, 4 cases were positive and the other 3 cases were negative by CC assay. It is concluded that YAC912C3 and interphase FISH providing a powerful technique in the detection of 20q(-) in MDS is an important complement to CC assay. PMID- 12744740 TI - [Preparation and radioimmunoimaging of a monoclonal antibody against human granulocytes]. AB - The aim of this research is to prepare a novel monoclonal antibody against granulocytes by the intraperitional routine procedure and evaluate the usefulness of (99)Tc(m) labelled anti-granulocyte monoclonal antibody (McAb) SZ-102 for the detection of experimental inflammatory areas in rabbits. It was characterized as IgG1 subclass by the double immunodiffusion analysis. The flow cytometry demonstrated that SZ-102 reacted selectively with human granulocytes, monocytes, and with their bone marrow precursors, while human lymphocytes, red blood cells and platelets remained negative. In addition, SZ-102 antigen was expressed on the macrophages of liver, lung, thymus, spleen and lymph node by immunohistochemical SP methods. It is suggested that McAb SZ-102 is mainly against granulocytes. SZ 102 was labelled with (99)Tc(m) using 2-iminothiolane modification McAb and (99)Tc(m) -glucoheptonate (GH) transchelation method. The experimental rabbit model of inflammatory areas was prepared,through injecting with (99)Tc(m)-SZ-102 by ear-edge vein, and then imaged by SPECT. (99)Tc(m) labelled murine IgG was used as a negative control. The inflammatory areas in rabbits were clearly imaged at 2 to 4 hour after injection of (99)Tc(m)-SZ-102, while the control group after injection of (99)Tc(m)-labeled murine IgG was negative. In conclusion, the McAb SZ-102 may be a potential agent for the diagnosis and localization of inflammatory areas of carcinomas and clinically concealed infectious diseases. PMID- 12744741 TI - [Preliminary study on HLA-B genotyping by oligonucleotide chips]. AB - HLA genes constitute a highly polymorphic multigene system. In the present study, HLA-B oligonucleotide chips were manufactured by using a set of sequence-specific oligonucleotide probes derived from polymorphic regions in exon 2 and exon 3 of HLA-B gene spotted by microarrayer onto the aldehyde modified glass slides. In addition, the sequenced HLA-B gene clones used as standard samples were amplified from exon 2 and exon 3 by PCR. Together with the correct hybridization and wash conditions, the PCR products were bound with the array probes on the chip, and the hybridization patterns were transformed to HLA-B genotypes. The results showed that the genotypes of standard samples by the HLA-B oligonucleotide chips were completely identical with the sequenced clones. In conclusion, the oligonucleotide chip method presented here for HLA-B genotyping is a rapid, accurate, sensitive and attractive high throughput biochemical way. PMID- 12744742 TI - [Application of double immunomagnetic positive sorting to ex vivo expansion of marrow CD34(+)CD59(+) cells from patients with paroxysmal nocturnal hemoglobinuria]. AB - Since flow cytometry was not feasible for sorting a huge amount of cells for clinical use, the method of double immunomagnetic positive sorting was used for selection of CD34(+)CD59(+) cells from bone marrow mononuclear cells in patients with paroxysmal nocturnal hemoglobinuria (PNH), which laid the groundwork for clinical ABMT/APBSCT of patients with PNH. Immunomagnetic positive selection was used for two times, the microbeads were removed from the CD34(+) cells selected firstly by means of overnight culture, then the sufficient CD34(+)CD59(+) cells were used for ex vivo expansion. The results showed that the survival, proliferation and colony-forming units of the selected CD34(+)CD59(+) cells by double immunomagnetic positive sorting had no significant difference as compared with that of CD34(+)CD59(+) cells selected by flow cytometry technique. It is suggested that the double immunomagnetic positive sorting promotes the use for separation and purification hematopoietic stem/progenitor cells and other cells with double or multiple markers cells for autologous hematopoietic stem cell transplantation in PNH patients. PMID- 12744743 TI - [A murine model of three mixed allogeneic bone marrow transplantation (A + B + C- >A)]. AB - In this paper, a model of three allogeneic mixed bone marrow transplantation of mice (BALB/c, H-2(d); C57BL/6, H-2(b); and CBA/N, H-2(k)) was established and whether or not prolongation of the survival time in recipient mice was observed. Lethally irradiated mice were transplanted with a mixture of a syngeneic plus two allogeneic bone marrow (A + B + C-->A, in a ratio of 1:4:4). At same time, mixed lymphocyte culture (MLC) with three allogeneic lymphocytes in vitro and two allogeneic antigens stimulated delayed type hypersensitivity (DTH) in vivo were performed. The results showed that the mice receiving mixed bone marrow transplant survived 56.6 +/- 27 days and the longest 103 days, however, only 15 +/- 5 days in the single allogeneic transplantation group. Whether two reacting cells to one stimulating cells or one reacting cells to two stimulating cells, all showed lower activity of MLC than that in one to one control group. In DTH assay, the mice sensitized with two allogeneic antigens showed lower reactivity than that in one antigen stimulated mice. It was concluded that the survival time of recipient mice was significantly prolonged after transplantation with three mixed allogeneic bone marrow. PMID- 12744744 TI - [Animal model of rat-to-mouse xenogeneic bone marrow transplantation with graft versus-host disease]. AB - To observe the graft-versus-host disease (GVHD) in rat-to-mouse model of bone marrow transplantation to build a GVHD model, BALB/c mice were conditioned with 8.5 Gy lethal total body irradiation and divided into two groups. One group of mice was infused with 4 x 10(7) bone marrow cells (BMC) from SD rats. The other group of mice was infused with 4 x 10(7) bone marrow cell and 2 x 10(7) spleen cells from SD rats. GVHD in mice of two groups were observed for 60 days. The results showed that mice in the group infused with only BMC mostly (80%) survived more than 60 days, but in the other group infused with mixed BMC and spleen cells, all mice died within 14 days and showed GVHD with pathologic evidence. In conclusion, to induce GVHD in rat-to-mouse bone marrow transplantation needs additional rat spleen cells. PMID- 12744745 TI - [Analysis on ultrastructure of lyophilized red blood cells]. AB - The objective of the present study was designed to evaluate lyophilized red blood cells of the ultrastructure. Blood was drawn from healthy adult. In group 1, sample was fresh blood; in group 2, sample was added 35% glycerine, stored at -80 degrees C for 24 hours; in group 3, red blood cells stored at 4 degrees C for 5 hours, then were lyophilized for 16 hour. The sample was resuspended for measurements of count and electron microscopy study. The result showed that lyophilized red blood cells possessed relative integrated structure, red blood cell recovery was 53%. The mean diameter, optical density and integral optical density of red blood cell were 4.7 +/- 0.4, 0.14 +/- 0.03 and 1.58 +/- 0.46 in group 1; 4.6 +/- 0.7, 0.14 +/- 0.02 and 2.35 +/- 0.64 in group 2; 4.4 +/- 0.4, 0.17 +/- 0.05 and 2.35 +/- 0.46 in group 3, respectively. There was no significant difference in lyophilized and frozen group, but there was significant difference in lyophilized group and normal group. In conclusion, human red blood cells could be successfully lyophilized and possess relative integrated structure. The mean diameter, optical density and integral optical density of lyophilized red blood cells were similar to that of cryopreservation red cells. PMID- 12744746 TI - [Micro-column gel indirect anti-globulin technique for screening and identification of irregular antibody]. AB - The purpose of this study was to design an antibody screening method based on the micro-column gel indirect anti-globulin technique (MGIAT), using pooled cells and plasma, by comparison with the conventional indirect anti-globulin technique (CIAT) combined with a two-stage papain technique, and to explore the feasibility of the use of plasma instead of serum as test material. The samples of blood recipients in our hospital were screened for irregular antibody using pooled test cells. Screening of the antibodies was identified both by MGIAT and CIAT combined papain technique respectively. The results showed that the irregular erythrocyte antibodies were detected in 20 cases from 5,000 recipients screened by MGIAT, using pooled cells, the positive rate was 0.4%. The specificity of 20 cases of irregular antibodies was as follows: 2 cases of anti-D, 8 cases of anti-E, 1 cases of anti-C, 2 cases of anti-c, 2 cases of anti-Mi(a), 2 cases of anti-Jk(a), 1 case of anti-Le(a) and 2 cases of anti-Fy(a). Antibody was detected from 19 cases using CIAT. Anti-Le(a) was detected with adding complement from Le(a-b-) person. Only 13 cases antibody were found by papain technique. It was concluded that irregular antibody screening by MGIAT using pooled cells can take place of the CIAT combining with papain technique in clinical application. Plasma is superior to serum in antibody screening test. PMID- 12744747 TI - [Study on the serum level of granulocyte-macrophage colony stimulating factor in some hematopathy patients]. AB - In order to explore the serum level of granulocyte-macrophage colony stimulating factor (GM-CSF) in hematopathy patients, radioimmunoassay was used to detect GM CSF level in serum from 163 cases of hematopathy, including 36 chronic aplastic anemia, 42 chronic granulocytic leukemia, 54 acute myeloid leukemia, 31 acute lymphocytic leukemia, and 40 healthy adults as control. The results showed that the serum GM-CSF level increased in chronic aplastic anemia patients, and significantly decreased in acute and chronic leukemia patients. In conclusion, these findings indicated that secreting level of GM-CSF is abnormal in patients with acute/chronic leukemias and chronic aplastic anemia. PMID- 12744748 TI - [Prognostic significance of peripheral blood cell and bone marrow megakaryocyte counts in patients with idiopathic thrombocytopenic purpura]. AB - To clarify the prognostic contribution of peripheral blood cell and bone marrow megakaryocyte counts in patients with idiopathic thrombocytopenic purpura, a series of data of 299 ITP patients including the counts of peripheral white blood cells and platelets, their increase potentials after treatment and the megakaryocytic counts on the bone marrow smears at diagnosis as well were collected and retrospectively analyzed to correlate with the disease development. The results showed that peripheral white blood cell and platelet counts at diagnosis were not associated with the prognosis, but positively associated with the increment of platelet counts after treatment. The cure rate reached up to 94.9% in the group with the platelet level restored to 100 x 10(9)/L in two weeks of therapy. The numerous megakaryocyte counts in bone marrow at diagnosis indicated good prognosis that the cure rate was up to 86.1% when the counts were more than 100 per 1.5 x 3 cm of smear. In conclusion, bone marrow examination on the quantity and quality of megakaryocyte would be critical for diagnosis and evaluation of the prognosis. Consecutive platelet counts during therapy is useful to estimate the disease development. PMID- 12744749 TI - [Diagnosis and treatment of invasive pulmonary aspergillosis in malignant hematological diseases-3 cases reported]. AB - Invasive pulmonary aspergillosis is difficult to diagnose and a critical ill with high mortality. In this paper, the diagnosis and treatment of invasive pulmonary aspergillosis complicated in 3 cases of hematological malignancy (2 acute leukemias and 1 MDS-RA) were retrospectively analysed. All patients had histories of hypoimmunity and were received prophylactic antifungal treatment. Pulmonary aspergillosis infection still occurred and confirmedly diagnosed by sputum examination. After 7 to 14 days of combination treatment of liposomal amphotericin B, itraconazole and flucytosine, 2 cases were cured and another showed effective. In conclusion, early diagnosis and treatment of invasive pulmonary aspergillosis are very critical and the therapeutic effectiveness of combined scheme with liposomal amphotericin B, itraconazole and flucytosine is very effective for pulmonary aspergillosis. PMID- 12744750 TI - [Clinical analysis of curative effect of rhG-CSF on chemotherapy-induced leukopenia in cancer patients]. AB - Myelosupression is the major dose-limiting toxicity in most chemotherapeutic drugs. To evaluate the curative effect of a domestic product of rhG-CSF on chemotherapy-induced leukopenia, 132 patients with malignancies were enrolled, including 80 patients with lung cancer, 35 patients with breast cancer, 10 patients with nasopharyngeal carcinoma, 3 patients with non-Hodgkin's lymphoma, 2 patients with gastric carcinoma and 2 patients with bone metastasis. Total of 528 cycles of chemotherapy were performed. The results showed that according the grade of leukopenia, the different daily doses of the domestic product of rhG-CSF were administered: 75 micro g/d for 3 days, 150 micro g/d for 4 days and 300 micro g/d for 5 days, in grade I-II, grade III and grade IV groups, respectively, the times of recovery to normal level of white blood cells were 2.5, 4.2 and 7 days in 3 groups, respectively. In conclusion, The Chinese product of rhG-CSF shortened the duration of leukopenia and accelerated the hematologic recovery, which shows only slight side effects, so that patients receive the optimal doses of chemotherapy and completed the planned schedule on time. PMID- 12744751 TI - [Improvements of lentiviral vector and its application in gene therapy of hematological diseases]. AB - As lentiviral vector holds the characteristics of higher transfection to non dividing cells, larger capacity of transfer gene fragments, long-term expression of therapeutic gene and lower rate of immunological response, therefore it becomes potential viral vector in gene therapy. Improvements of lentiviral vector, human immunodeficiency virus type-I as example, and its application in gene transfer for gene therapy of hematological diseases are emphasized in this review. PMID- 12744753 TI - Why aren't we doing better? PMID- 12744752 TI - [The approaches of chronic graft-versus-host disease]. AB - Chronic graft-versus-host disease (cGVHD) continues to be a major complication in long-term survivors after allogeneic hematopoietic stem cell transplantation and is the principal cause of morbidity and non-relapse mortality. With the new approaches in the immune mechanisms of cGVHD, the diagnosis, prophylaxis and treatment of cGVHD are involved. This review summarises the current progress of research on cGVHD. PMID- 12744756 TI - Trastuzumab-based combinations in metastatic breast cancer: how to make a choice. AB - Trastuzumab, a humanized monoclonal anti-human epidermal growth factor receptor-2 (HER2) antibody that has proven efficacy as monotherapy in HER2-overexpressing breast cancer, has a favorable toxicity profile characterized by infrequent infusion reactions and a slightly increased incidence of cardiac dysfunction. Regimens that combine trastuzumab with a cytotoxic agent can increase the overall response rate and prolong survival in patients with metastatic breast cancer. Trastuzumab has been investigated in combination with anthracyclines, taxanes, platinum salts, vinorelbine, gemcitabine, capecitabine, and combinations of these agents. Combining trastuzumab with these agents enhances the toxicity of treatment and can especially enhance the risk of cardiotoxicity. This article summarizes data on trastuzumab-based combinations with particular attention to the risk benefit ratio of these therapies. HER2 testing by immunohistochemistry or fluorescence in situ hybridization is discussed in the context of determining the optimal course of treatment for patients. PMID- 12744757 TI - Nonmyeloablative allogeneic hematopoietic stem cell transplantation for metastatic breast cancer. AB - Previously, there was very little interest in investigating allogeneic hematopoietic stem cell transplantation (alloHSCT) in breast cancer because of the significant morbidity and mortality associated with this procedure, as well as the disappointing results observed in clinical trials with high-dose chemotherapy and autologous hematopoietic stem cell transplantation in advanced breast cancer. However, the development of nonmyeloablative (reduced-intensity) conditioning regimens, which have less treatment-related mortality but preserve the T cell-mediated graft-versus-tumor (GVT) effect, has led to the investigation of nonmyeloablative alloHSCT in diseases that had not previously been considered for conventional alloHSCT, including metastatic breast cancer. Laboratory data demonstrate that T cell-mediated responses to breast cancer that inhibit tumor growth are possible and provide the rationale to pursue allogeneic adoptive cellular therapy as a strategy to eliminate breast cancer. Early reports of nonmyeloablative alloHSCT indicate that a clinical GVT effect against breast cancer does exist. The responses appear to be dependent on the development of complete donor lymphoid chimerism, and responses may be delayed. The results from these initial trials must be interpreted cautiously. It is unlikely that nonmyeloablative alloHSCT by itself will result in complete eradication of metastatic breast cancer; however, it may serve as a therapeutic platform to enhance the effects of currently available immunotherapies (eg, trastuzumab administration) and complement existing cytotoxic therapies. Well-designed studies will be necessary to determine the clinical efficacy of nonmyeloablative alloHSCT as adoptive cellular therapy in metastatic breast cancer. PMID- 12744758 TI - Gemcitabine/vinorelbine in metastatic breast cancer patients previously treated with anthracyclines: results of a phase II trial. AB - This phase II study was designed to evaluate the response rate (RR) and toxicity of gemcitabine/vinorelbine in patients with metastatic breast cancer. All patients had previously received anthracyclines. Treatment consisted of gemcitabine 1200 mg/m2 and vinorelbine 30 mg/m2 on days 1 and 8, every 3 weeks. Twenty-five patients were enrolled. Median age was 59 years (range, 33-73 years). Ten patients had received only adjuvant therapy with anthracyclines. The remaining 15 patients had received chemotherapy for metastatic disease, including taxanes in 11 cases. Four patients could not be evaluated for response. By intent to-treat analysis, the overall RR was 44% (95% CI, 24.4%-65%). Median duration of response and median time to treatment failure were 21 and 17 weeks, respectively. The main toxicity was hematologic, with grade 3/4 neutropenia occurring in 13 patients and 1 patient developing febrile neutropenia. Two deaths from pneumonia occurred. These results reveal an encouraging activity with a reasonable toxicity profile in a patient population with an unfavorable prognosis. Our group is conducting a randomized study to compare this combination with vinorelbine alone in patients with metastatic breast cancer after failure to respond to anthracyclines and taxanes PMID- 12744759 TI - Metastasis suppressor genes: basic biology and potential clinical use. AB - Metastatic disease remains a significant contributor to morbidity and mortality in patients with breast cancer. An improved molecular and biochemical understanding of the metastatic process is expected to fuel the development of new therapeutic approaches. The suppression of tumor metastasis, despite tumor cell expression of oncogenes and metastasis-promoting events, has become a diverse and fruitful field of investigation. Although many genetic events promote metastasis, several genes show relatively reduced expression levels in metastatic tumor cells in mouse model systems and in aggressive human tumors. Re-expression of a metastasis-suppressor gene in a metastatic tumor cell line results in a significant reduction in metastatic behavior in vivo with no effect on tumorigenicity. The known metastasis-suppressor gene products nm23, KAI1, mitogen activated protein kinase kinase 4, breast cancer metastasis suppressor-1, KiSS1, RHOGDI2, CRSP3, and vitamin D3-upregulated protein/thioredoxin interacting protein exhibit unexpected biochemical functions that have shed new light on signaling events that are important in metastasis. Most metastasis suppressors function at the translationally important stage of outgrowth of micrometastatic tumor cells at a distant site. We hypothesize that elevation of metastasis suppressor gene expression in micrometastatic tumor cells in the adjuvant high risk population of patients with breast cancer will halt metastatic colonization and have a clinical benefit. DNA methylation inhibitors have shown limited promise in increasing metastasis-suppressor gene expression, and ligands of the nuclear hormone receptor family are currently under investigation in vitro and in vivo. Clinical testing of agents that increase metastasis-suppressor gene expression is expected to require tailored trial designs. PMID- 12744760 TI - Digital mammography: a review of technical development and clinical applications. AB - For detecting and diagnosing breast cancer at its earliest stage, mammography is the most sensitive technique currently available and is therefore the method of choice. Screen-film mammography has been used successfully as a screening test for breast cancer for > 2 decades. However, conventional mammography has substantial limitations and, therefore, digital mammography systems have been developed to improve image quality and overcome the limitations of screen-film technique limitations. Herein we discuss the differences between screen-film and digital mammography systems and the processes related to digital mammography that differ from conventional mammography, including detector technology, digital image formation, image processing, image display, and image archival. Finally, we review the results from currently available clinical trials regarding the performance of digital mammography and discuss clinical implications such as cost effectiveness. PMID- 12744761 TI - Mononuclear cell function and energetics in early traumatic injury. AB - BACKGROUND: The reason for increased infectious complications after traumatic injury is complex and incompletely understood. We propose a relationship between the energetic state of circulating immune cells and immune cell function in traumatic injury. To examine this relationship, cellular adenosine triphosphate (ATP) concentration and cellular functions were measured in peripheral blood mononuclear cells from trauma patients and normal subjects. MATERIALS AND METHODS: Mononuclear cells were isolated within 24 h of injury from trauma patients without (n = 12) or with (n = 10) hypotension (groups 1 and 2, respectively), and a group of normal control subjects (group 3, n = 13). Mononuclear cells were assayed for ATP levels using bioluminescence. Phagocytosis was quantified via flow cytometry after ingestion of fluorescent microspheres and phagocytic index (PI) was calculated (average number of particles ingested per monocyte). Protein synthesis was quantified using incorporation of (35)S-labeled methionine into cultured cells. Comparisons between groups were performed using one-way analysis of variance (ANOVA) with adjustment for multiple comparisons. RESULTS: All but one trauma patient suffered blunt injury. There were significantly more transfusions in group 2 (hypotensive) patients (p = 0.0005). Nosocomial infections, length of stay (LOS), and mortality did not differ between groups 1 and 2. Concentrations of ATP and PI in both groups of traum a patients did not significantly differ from controls. In hypotensive trauma patients, increased duration of hypotension was associated with increased mononuclear cell ATP levels (r(2) = 0.227). A negative correlation between PI and ATP levels in trauma patients was discovered. Incorporation of (35)S was significantly greater in normotensive trauma patients than controls. CONCLUSION: Mononuclear cell ATP levels and measured functions are preserved in early traumatic injury. PMID- 12744762 TI - Microdialysis: a new technique to monitor perioperative human peritoneal mediator production. AB - BACKGROUND: Standardized methods to measure peritoneal cytokine production do not exist. This feasibility study examines the use of microdialysis to monitor perioperative peritoneal mediator production in patients following abdominal surgery for infective or non-infective conditions. MATERIALS AND METHODS: At the beginning of the operation, a microdialysis catheter was placed between the patient's parietal peritoneum and the muscular fascia of the abdominal wall in the connective tissue bed. The device was irrigated (18 microL/h, Ringer's solution/0.05% albumin) for up to 7 days. Samples of the dialysate were collected at least twice a day, and concentrations of interleukin (IL)-6 and monocyte chemoattractant protein (MCP)-1 were measured by an ELISA technique. Four of the nine patients included had proved intra-abdominal infections. RESULTS: In uninfected patients, IL-6 concentrations peaked 8 h after skin incision (mean +/- SEM): 1696 +/- 1292 pg/mL and dropped rapidly to significantly lower concentrations (less than 400 pg/mL) thereafter. MCP-1 concentrations also peaked at 8 h (12787 +/- 6893 pg/mL). In the following days, MCP-1 concentrations were variable between 1000 and 5000 pg/mL. In infected patients, early IL-6 production tended to be higher and that of MCP-1 tended to be lower than in uninfected patients. Catheters were removed between day four and day seven when the system failed or when the patients became mobile without any clinical symptoms of complications. CONCLUSION: The samples derived from microdialysis were suitable to measure sub-peritoneal mediator profiles during surgery and up to 7 days postoperatively. Microdialysis data should be validated for a potential correlation with the clinical course. PMID- 12744763 TI - Sublethal hemorrhage induces tolerance in animals exposed to cecal ligation and puncture by altering p38, p44/42, and SAPK/JNK MAP kinase activation. AB - BACKGROUND: We have shown that SLH induces tolerance to endotoxin in vivo and in vitro, and is associated with alterations in MAP kinase (p38, p44/42, and SAPK/JNK) activation and TNF production. This study investigates the effect of sublethal hemorrhage (SLH) on cecal ligation and puncture (CLP) polymicrobial sepsis and examined the effect of the bioflavinoid, curcumin, a MAP kinase inhibitor, on this relationship. MATERIALS AND METHODS: Sprague-Dawley rats underwent SLH (hemorrhage and MAP = 30 mm Hg for 15 min, with shed blood returned) or sham operation. After 24 h, rats had CLP (cecal base ligation with double puncture). Survival was determined +/- curcumin pretreatment (n = 10/group). Lung tissue, serum, and bronchoalveolar lavage (BAL) fluid were obtained 30 min after SLH and 4 and 12 h after CLP (n = 8/group). Lung tissue was analyzed for p38, p44/42 SAPK/JNK, and HSP-70 phosphorylation (Western). Lung myeloperoxidase (MPO) activity was measured as an index of neutrophil infiltration. TNF ELISA was performed on serum and BAL sample. RESULTS: SLH significantly improved survival after CLP (21.5 vs. 7.5 h vs. sham, p = 0.008), and curcumin reversed this benefit (7.3 h, p = 0.0007 vs. SLH + CLP). MAP kinase activity was significantly greater in SLH rats 4 h post-CLP (p38: 720 vs. 331, p = 0.03, p44/42: 2759 vs. 1295, p = 0.006, SAPK: 413 vs. 254). Curcumin significantly inhibited MAPK activity both 30 min after SLH (p38: 297 vs. 3260, p44/42: 370 vs. 2628, SAPK: 748 vs. 1764, all p < 0.01 vs. SLH 30 min) and 4 h post CLP (p38: 146 vs. 720, p44/42: 616 vs. 2759, all p < 0.01 vs. SLH + CLP4 h). Four hours after CLP, SLH rats expressed more HSP72. Lung MPO activity was significantly lower in SLH + CLP rats at both 4 h (9.5 vs. 15.6, p = 0.02 vs. sham) and 12 h (18.1 vs. 37.5, p = 7 x 10(-5), vs. sham). Serum and BAL TNF levels were higher in SLH rats initially (serum: 145 vs. 28 pg/mL, p = 2 x 10(-5) BAL: 83 vs. 57 vs. sham + CLP4h); however, BAL TNF was significantly lower in SLH animals 12 h post-CLP (37 vs. 72.7 pg/mL, p = 0.003 vs. sham + CLP12h). CONCLUSION: SLH induces tolerance to CLP. This tolerance is dependent on early MAP kinase activation, since the survival benefit is reversed by curcumin. Decreases in tissue cytokine levels and neutrophil infiltration are potential mechanisms by which SLH induces tolerance to CLP (polymicrobial sepsis), attenuates acute lung injury, and improves survival. PMID- 12744764 TI - Pathogenesis of posttraumatic empyema: the impact of pneumonia on pleural space infections. AB - BACKGROUND: Thoracic empyema may result either from primary pneumonic sources or intraabdominal sources of infection that seed the pleural space secondarily. In patients with thoracostomy tubes, empyema may result when blood in the pleural space becomes contaminated during tube insertion. To elucidate the cause of posttraumatic empyema, preoperative bronchoalveolar lavage (BAL)/sputum cultures obtained from patients with posttraumatic empyema were compared with cultures obtained at the time of decortication. MATERIALS AND METHODS: A retrospective study was conducted of trauma patients who developed empyema and underwent either video-assisted thoracoscopy or thoracotomy with decortication following blunt or penetrating trauma. At our level I trauma center, we studied all empyema cases diagnosed from November, 1998 to July, 2001. Data collection included patient demographics, injuries sustained, preoperative BAL/sputum cultures, and culture data obtained at the time of decortication. All BAL/sputum cultures were performed no more than 5 days prior to decortication. RESULTS: Thirty-seven patients (26 blunt/11 penetrating) were identified. No patients had concurrent intra-abdominal sources of infection. All patients had at least one chest tube placed prior to decortication. Preoperative respiratory cultures (BAL/sputum) were obtained in 34 patients. The most common organisms isolated were Staphylococcus aureus in six patients (18%) and Hemophilus influenzae in six patients (18%). Intraoperative cultures were obtained in all 37 patients, with the most common organism being S. aureus isolated in 22 patients (60%). Interestingly, a correlation between preoperative BAL/sputum and intraoperative cultures was found in only seven of the 34 patients (21%) who had concomitant respiratory and pleural cultures. Cultures positive for S. aureus were isolated from five patients, Streptococcus pneumoniae from one patient, and Pseudomonas aeruginosa from one patient. CONCLUSION: Little correlation existed between preoperative BAL/sputum cultures and intraoperative cultures in this series of patients with posttraumatic empyema. This suggests that the causation is most often not a parapneumonic process. Furthermore, since S. aureus was the most common organism recovered from empyema, the source was more likely from inoculation of the pleural space by the injury itself or by tube thoracostomy. PMID- 12744765 TI - Sexual dimorphism in the activation of neutrophils by shock mesenteric lymph. AB - BACKGROUND: After trauma/hemorrhagic shock (T/HS), inflammatory products exit the gut via mesenteric lymph. These products can prime neutrophils (PMN) and predispose to lung injury. Female gender and the proestrous state (PE) may confer protection against lung injury after T/HS. We therefore studied the dependence of T/HS-induced PMN priming on gender and stage of estrous. MATERIALS AND METHODS: T/HS was induced in male (M) and proestrous female (F) rats by laparotomy plus hemorrhagic shock (30 mm Hg, 90 min) followed by reinfusion of shed blood. Six hours later rats were sacrificed and plasma was obtained. Control male rat PMN were primed 5 min in buffer or in the plasma of M-T/HS or F-T/HS rats (n = 4 6/group). PMN were then assayed using DHR for respiratory burst (RB) initiated by sequential MIP-2 and PAF stimulation (MIP/PAF). Because MIP and PAF mobilize cell calcium ([Ca(2+)](i)) in a step crucial for RB initiation, we also assayed PMN [Ca(2+)](i) responses to MIP/PAF. RESULTS: M-T/HS plasma primed PMN RB (208 +/- 8 [SEM] U/sec versus buffer 51 +/- 12 U/sec, p < 0.01, ANOVA/Tukey's). F-T/HS plasma did not (87 +/- 20 U/sec, NS). PMN basal [Ca(2+)](i) was increased by pre incubation in both M-T/HS and F-T/HS plasma (183 +/- 26 and 225 +/- 20 nM, p < 0.02, p < 0.01 compared to buffer [80 +/- 3 nM]). Peak PMN [Ca(2+)](i) response to MIP/PAF was 159 +/- 2 nM without priming. Priming PMN in either M-T/HS or F T/HS plasma increased peak [Ca(2+)](i) responses to MIP/PAF to 274 +/- 35 (p < 0.04) and 330 +/- 24 nM (p < 0.02), but the effects of M-T/HS and F-T/HS plasma on [Ca(2+)](i) mobilization were indistinguishable. CONCLUSION: Plasma from male rats subjected to T/HS primes PMN respiratory burst, but plasma from proestrous females subjected to T/HS does not. In contrast, the male and proestrous female plasma primed PMN [Ca(2+)](i) mobilization by MIP/PAF equally. The decrease in pathologic PMN activation seen after T/HS in proestrous female rats depends on soluble mediators present in plasma. The decreased PMN RB seen after T/HS in proestrous females is mediated by calcium-independent pathways. PMID- 12744766 TI - Infections in 346 consecutive video-assisted thoracoscopic procedures. AB - BACKGROUND: Postoperative infections, as related to risk factors, in patients undergoing video-assisted thoracoscopic surgery (VATS) procedures have been studied infrequently. MATERIALS AND METHODS: We evaluated 346 consecutive patients who underwent VATS procedures between October 1996 and June 2002 at our center. Patients preoperatively were free of chest infections and were divided into two groups: Group A (n = 139) who underwent lung wedge resection; group B (n = 207), who underwent pleural biopsy (n = 183) or biopsy of a mediastinal mass (n = 24). We recorded prospectively the following preoperative infection risk parameters: Hemoglobin concentration, hematocrit, serum albumin concentration, lymphocyte count, length of preoperative stay, duration of surgery, blood transfusion, age, comorbidity, and chronic obstructive pulmonary disease specifically (COPD, measured as FEV(1) <70% of expected). Short-term antibiotic prophylaxis was given to 94% of patients in group A and to 90% of patients in group B. As outcome measures we recorded the occurrence of postoperative infections within 30 days (surgical site infection, pneumonia, empyema) and the final patient outcome. RESULTS: Patients who developed postoperative infections (all the above types included) were 17/346 (4.9%), the difference between group A (5.0%) and group B (4.8%) being not significant. The overall surgical site infection rate was 1.7%. Groups A and B showed a similar incidence of surgical site infection (2.8% vs. 1.0%; p = NS), of pneumonia (2.8% vs. 3.4%; p = NS), and of empyema (0.7% vs. 2.0%; p = NS). Among assessed infection risk parameters, a FEV(1) <70% of expected was the only parameter associated with a significantly increased incidence of surgical site infection (p < 0.05). CONCLUSIONS: This prospective study confirms that the wound infection rate is low (1.7%) after minimally invasive VATS procedures. The cumulative incidence of postoperative infections (including wound infection, pneumonia, empyema) was similar after lung wedge resection and after pleural or mediastinal mass biopsy procedures. Among the infection risk parameters, COPD was the only parameter associated with a significantly increased incidence of postoperative infection. Our results suggest that patients with COPD who undergo VATS for lung wedge resections and for pleural/mediastinal biopsy should receive antibiotic prophylaxis to prevent surgical site infection. PMID- 12744767 TI - Eradication of methicillin-resistant Staphylococcus aureus from pressure sores using warming therapy. AB - BACKGROUND: Nosocomial infection is a major cause of surgical morbidity and mortality. Methicillin-resistant Staphylococcus aureus (MRSA) has become a prominent organism in colonization and infection in surgical patients. Pressure sores are a major reservoir of MRSA. MATERIALS AND METHODS: In this study, 33 patients with full-thickness pressure sores were randomized to receive standard care or radiant heat therapy using a Warm Up device (Augustine Medical, Eden Prairie, MN). Weekly microbial sampling was used for assessment of bacterial presence. None of the patients received antibiotics prior to or during the eight weeks of study. RESULTS: More than 50 species of bacteria were present in the pressure sores with a median of four organisms per sample. Methicillin-resistant S. aureus was found in 14 of the patients' pressure sores. In the warming group (n = 8), MRSA was eradicated in six patients within 2 weeks of warming, whereas in the control group none had eradication (Fisher's exact test, p = 0.01). Eradication was defined as three consecutive weekly swabs without bacterial growth. CONCLUSION: The warming of pressure sores is being assessed as an adjunct to healing, but there is some promise that colonization by MRSA may be eradicated, thereby reducing a potential reservoir of organisms. The risk to surgical patients when patients are harboring MRSA may be minimized by warming therapy. PMID- 12744768 TI - Effect of linezolid versus vancomycin on length of hospital stay in patients with complicated skin and soft tissue infections caused by known or suspected methicillin-resistant staphylococci: results from a randomized clinical trial. AB - BACKGROUND: Complicated skin and soft tissue infections are common surgical indications usually requiring patients to be hospitalized, and are often caused by gram-positive bacteria, including methicillin-resistant staphylococci such as MRSA. Vancomycin has been the standard treatment for methicillin-resistant staphylococcal infections in many countries, but its intravenous-only formulation for systemic infections often confines patients to the hospital for the treatment. Linezolid, a novel oxazolidinone antibiotic available in intravenous and 100% bioavailable oral forms, was shown in a randomized trial to be as efficacious as vancomycin for suspected or proven methicillin-resistant staphylococcal infections. To determine if oral linezolid can reduce length of hospital stay (LOS) when compared to vancomycin, we compared the LOS for the 230 complicated skin and soft tissue infection patients enrolled in this trial. MATERIALS AND METHODS: Patients received up to four weeks of linezolid (intravenous followed by optional oral) or vancomycin (intravenous only), followed by up to four weeks of observation. Unadjusted LOS was estimated using Kaplan-Meier survival functions, whereas the log-logistic survival analysis model was used to estimate the multivariate-adjusted LOS controlling for patient demographics and selected baseline clinical variables. Analysis was done on the intent-to-treat (n = 230) sample as well as on two subsamples of the clinically evaluable (n = 144) and surgical site infection (n = 114) patients. RESULTS: The unadjusted Kaplan-Meier median LOS was five days shorter for the linezolid group than the vancomycin group in the intent-to-treat sample (9 vs. 14 days, p = 0.052). It was eight days shorter (8 vs. 16 days, p = 0.0025) in the clinically evaluable sample, but the difference in the surgical site infection sample was not significant (10 vs. 14 days; p = 0.29). The linezolid group's unadjusted mean LOS was 1.7, 5.3 and 0.8 days shorter in the intentto-treat, clinically evaluable, and surgical site infection samples, respectively. After adjusting for age, gender, race, geographic region, bacteremia, type of inpatient location, and number of concurrent medical conditions using the log-logistic model, between treatment differences in the multivariate-adjusted median LOS decreased to 3, 6, and 3 days, whereas the differences in mean LOS increased to 3.1, 6.5 and 2.5 days for the intent-to-treat, clinically evaluable, and surgical site infection samples (p < 0.01, < 0.01, and < 0.10), respectively. When the between-treatment differences in LOS were expressed as odds ratio of hospital discharges, multivariate-adjustment increased the odds ratios in favor of linezolid for all the three samples. CONCLUSION: Results from this randomized trial show that linezolid can significantly reduce LOS for patients with complicated skin and soft tissue infections from suspected or confirmed methicillin-resistant staphylococci. PMID- 12744769 TI - Society profiles. Interview by Vicki Glaser. PMID- 12744771 TI - GM-CSF induction in human lung fibroblasts by IL-1beta, TNF-alpha, and macrophage contact. AB - Fibroblast-derived cytokines may play crucial roles in airway inflammation. In this study, we analyzed expression of the inflammatory cytokine, granulocyte macrophage colony-stimulating factor (GM-CSF), a major eosinophilopoietin, by normal human lung fibroblast (NHLF) cells and its regulation by monokines and macrophage contact. NHLFs were stimulated with interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) and were cocultured with the U937 myelomonocytic cell line. The expression of GM-CSF transcripts was analyzed by reverse transcription-polymerase chain reaction (RT-PCR), and GM-CSF protein was detected by ELISA. Nuclear translocation of nuclear factor-kappaB (NF-kappaB), an important transcription factor for inflammatory gene expression, was assessed by electrophoretic mobility shift assay (EMSA). Both IL-1beta and TNF-alpha significantly enhanced the production of GM-CSF by NHLF. Coculturing of peripheral blood mononuclear cells (PBMC) with NHLF induced GM-CSF expression. This phenomenon was also seen on coculturing U937 cells or membranes derived from U937 with NHLF but was inhibited when the two types of cells were separated, suggesting a need for cell-cell contact. U937 membranes, as well as IL-1beta and TNF-alpha, induced nuclear translocation of NF-kappaB. These data support a prominent role for macrophage-fibroblast interactions in airway inflammation and fibrosis. PMID- 12744772 TI - CD2-CD48 interactions promote cytotoxic T lymphocyte induction and function: anti CD2 and anti-CD48 antibodies impair cytokine synthesis, proliferation, target recognition/adhesion, and cytotoxicity. AB - The role of CD2 signaling in cytotoxic T lymphocyte (CTL) development was examined by stimulating mouse T cells with anti-CD3 monoclonal antibody (mAb) in the absence or presence of anti-CD2 mAb or anti-CD48 mAb or both. Induction of nonspecific CTL and interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) synthesis were impaired in the absence of CD2-CD48 interactions. Anti-CD2 mAb also inhibited activation-induced expression of the high-affinity IL-2 receptor (IL-2R). In contrast, IFN-gamma receptor (IFNGR) expression was increased in the presence of anti-CD2 mAb. Reduced cytotoxicity by CTL induced in the absence of CD2-CD48 interactions was associated with a diminished ability of CTL to conjugate with target cells and reduced expression of granzyme B and perforin. Anti-CD2 mAb did not affect expression of Fas ligand and tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) by anti-CD3-activated T cells. Cytotoxic effector function and granzyme B and perforin expression were rescued when exogenous IL-2 and IFN-gamma were added in combination with anti-CD2 mAb to anti-CD3-activated T cells at initiation of culture. We conclude that CD2-CD48 interactions during T cell activation are critical for the synthesis of sufficient IL-2 and IFN-gamma to drive CD8(+) T cells to differentiate into functional cytotoxic effector cells. PMID- 12744773 TI - IFN-beta1a (Rebif) modifies the expression of microfilament-associated cell-cell contacts in C6 glioma cells. AB - Multiple sclerosis (MS) is a chronic inflammatory disease characterized by multifocal demyelination and axonal damage in the central nervous system (CNS). The disruption of the endothelial blood-brain barrier (BBB) with consecutive transmigration of inflammatory cells into the brain parenchyma is of critical importance in the pathogenesis of MS. The integrity of the BBB and the adjacent network of glial cells partially depends on the assembly of intercellular contacts between astrocytes. We demonstrate that recombinant interferon-gamma (rIFN-gamma), a proinflammatory cytokine critically involved in the disruption of the BBB, downregulates the expression of the cell adhesion molecules N-cadherin and vinculin in astrocytic C6 cells using Western blot and immunofluorescence microscopy. By contrast, IFN-beta1a, an established treatment for relapsing remitting MS, increases the expression of N-cadherin and vinculin and partly inhibits the downregulation of these adhesion molecules by phytohemagglutinin (PHA)-stimulated IFN-gamma-secreting human T lymphocytes in coculture experiments. In summary, we demonstrate that IFN-beta1a modifies the assembly of N-cadherin- and vinculin-mediated intercellular contacts between astrocytic C6 cells in vitro. This effect may also contribute to the therapeutic action of IFN beta1a in MS. PMID- 12744774 TI - Individual variability in cyclosporin A sensitivity: the assessment of functional measures on CD28-mediated costimulation of human whole blood T lymphocytes. AB - The quantitative analysis of cyclosporin A (CsA) effects might be helpful for optimizing immunosuppressive treatment after allogeneic organ transplantation in individual patients, as rejection can occur despite the existence of CsA blood levels within therapeutic ranges. Previous investigations found that costimulation of the CD28 pathway generally mediates CsA-resistant proliferation of T cell receptor (TCR)-activated T lymphocytes. However, here we describe considerable interindividual variation regarding the immunosuppressive effects of CsA (1000 microg/L) on anti-CD3/CD28 T cell costimulation in a human whole blood assay. In the in vitro study, we found a significant reduction of T cell proliferation, activation marker expression (CD25, CD69) on the T cell surface, and interleukin-2 (IL-2) protein expression in whole blood samples of all healthy subjects (n = 11). However, the investigation of cytokine mRNA profiles revealed variable results of in vitro CsA sensitivity. Whole blood samples of 3 of 11 healthy individuals demonstrated a marked suppression of IL-2 mRNA expression (>50%) and a partial inhibition of IL-4, interferon-gamma (IFN-gamma), and tumor necrosis factor-alpha (TNF-alpha) mRNA expression on addition of CsA. In contrast, the remaining 8 healthy individuals had cytokine mRNA expression levels that were unaffected or even increased when CsA was administered in vitro. In patients undergoing CsA monotherapy (ex vivo study, n = 9), we found a significant suppression of IL-2 mRNA levels in 4 of 9 patients ex vivo. Thus, we cannot confirm a universal CsA resistance of T cells on anti-CD3/CD28 costimulation. Instead, our results suggest an individual degree of CsA sensitivity that might be more consistent with clinical experience. Prospective studies are necessary to determine if individual degrees of CsA sensitivity correlate with clinical events and are associated with a low or high risk of transplant rejection. PMID- 12744775 TI - Cloning and characterization of porcine common gamma chain gene. AB - The common gamma chain, which was originally identified as a component of interleukin-2 receptors (IL-2R), plays a key role in differentiation of T lymphocytes and natural killer (NK) cells. In the present study, cDNA of the porcine common gamma chain gene and its genomic DNA were molecularly cloned and characterized. The porcine common gamma chain gene was found to consist of 8 exons, spanning approximately 3.7 kb, and to encode a 368-amino acid polypeptide. The amino acid sequence showed 82.4%, 71.1%, 86.1%, and 84.8% similarities with that of human, murine, bovine, and canine chains, respectively. The common gamma chain gene was assigned to swine chromosome Xq13 by FISH analysis and was consistent with the result of radiation hybrid (RH) mapping. When various porcine tissues were examined for the expression of this gene, the expression was observed in lymphocytes and lymphocyte-related tissues. Since GATA, T cell factor 1 (TCF-1), Ets-1, activated protein2 (AP-2), and Ikaros2 binding motifs were demonstrated in the 5' upstream region of this gene, promoter activity was investigated using luciferase gene as a reporter. The results indicate that the Ets-1 binding motif in the segment from -95 to -59 (major transcription initiation site: +1) was an essential cis-acting regulatory element for the common gamma chain gene in lymphoid cells. PMID- 12744776 TI - The primary role in biologic activity of the neutrophil chemokines IL-8 and GRO alpha in cultured nasal epithelial cells. AB - Primary nasal epithelial cells were investigated for their ability to synthesize and deliver neutrophil chemotactic factors (chemokines) following tumor necrosis factor-alpha (TNF-alpha) induction. The chemokines interleukin8 (IL-8), growth related oncogene-alpha (GRO-alpha), epithelial cell-derived neutrophil attractant 78 (ENA-78), and granulocyte chemotactic protein-2 (GCP-2) have been detected and characterized and shown to have different potencies in the chemotaxis of neutrophils. Cultures of primary nasal epithelial cells were treated with TNF alpha in concentrations of 20 and 200 ng/ml for 2, 8, 24, and 72 h. The chemokine protein concentrations in the supernatants of the incubations were determined by the ELISA technique. Chemokine mRNA expression in epithelial cells was also measured using the reverse transcriptase-polymerase chain reaction (RT-PCR). The biologic activity of the chemokines was identified using a three-step high performance liquid chromatography (HPLC) technique, a bioassay involving measurement of neutrophil chemotaxis in a single Boyden chamber. Both the IL-8 and GRO-alpha proteins and their respective mRNA appear to be induced by TNF alpha in epithelial cells. The chemotactic responsiveness of both GRO-alpha and IL-8 appears to predominate after 24 h incubation with TNF-alpha. The chemokines GCP-2 and ENA-78 were only weakly induced by TNF-alpha. The neutrophil chemokines IL-8 and GRO-alpha were synthesized in nasal epithelial cell culture induced by TNF-alpha in biologically active concentrations of 0.8 ng/ml and 1.42 ng/ml, respectively. It appears that both the IL-8 and GRO-alpha chemokines may contribute to neutrophil tissue migration in sinusitis, whereas GCP-2 and ENA-78 are of secondary importance to the chemotaxis of neutrophils in this condition. PMID- 12744777 TI - [Ensuring safe blood in the Americas]. PMID- 12744778 TI - [Promoting quality blood services in the Region of the Americas]. PMID- 12744781 TI - Blood services in the Region of the Americas. PMID- 12744783 TI - Basic components of a national blood system. PMID- 12744784 TI - [A study of sociocultural factors related to voluntary blood donation in the Americas]. AB - OBJECTIVE: To obtain baseline data for countries of the Americas on knowledge, attitudes, and practices related to voluntary blood donation as well as on the current level and quality of services that blood banks provide to donors. METHODS: The study was conducted in 15 countries in the Americas: Argentina, Bolivia, Colombia, Costa Rica, Cuba, the Dominican Republic, Ecuador, El Salvador, Guatemala, Jamaica, Nicaragua, Panama, Paraguay, Peru, and Venezuela. Technical cooperation for the study came from the Pan American Health Organization. A qualitative formative methodology was applied, utilizing interviews with donors, health workers, and members of the general public; direct observation; focus groups; knowledge tests; and a review of documents. RESULTS: Information was generated on people's knowledge of donation; their beliefs, perceptions, attitudes, and motivations; and their barriers to donating. Knowledge was also gained as to the best means for disseminating messages supporting voluntary donation. CONCLUSIONS: This information will serve as a foundation for designing a strategy in the countries of the Americas that is aimed at establishing and building the loyalty of voluntary blood donors. This strategy can support the implementation of changes needed in the care of donors, and it can also help in motivating donors to regularly return to donate blood. PMID- 12744785 TI - Availability, safety, and quality of blood for transfusion in the Americas. AB - OBJECTIVES: This article has two objectives: (1) to present for countries and territories of the Region of the Americas data on the number of blood donations, proportion of voluntary blood donors versus remunerated blood donors, coverage of screening for infectious agents, and separation of donated blood into its components and (2) to explore the relationships of those characteristics with economic and organizational factors in the countries and territories. METHODS: We carried out comparative analyses using population and health information gathered annually by the Pan American Health Organization (PAHO) from national health officials from the countries in the Americas, as well as economic information (gross national product (GNP) per capita) obtained from publications of the World Bank. RESULTS: There is a direct correlation between the availability of blood for transfusion and GNP per capita. Seven countries with a GNP per capita above US$ 10 000 per year account for 38% of the Regional population but 68% of the Regional blood donations. Voluntary blood donation is more common in the countries with better blood availability. There is no association between GNP per capita and coverage of screening for infectious agents. Nevertheless, of the six countries with a GNP per capita below US$ 1 000, only one of the six screens all units for human immunodeficiency virus (HIV), hepatitis C virus (HCV), and hepatitis B surface antigen (HBsAg). Countries with a higher proportion of voluntary blood donors tend to have lower prevalence rates of infectious markers. Separation of blood into its components is also more common in countries with higher blood donation rates. CONCLUSIONS: The availability, safety, and quality of blood for transfusion in the Americas needs to be improved. As part of that effort, national policies and strategies must be put into place so that the resources already allocated for blood services are better utilized. PMID- 12744786 TI - [The discarding of blood units and the prevalence of infectious diseases in donors at the Pro-Blood Foundation/Blood Center of Sao Paulo, Sao Paulo, Brazil]. AB - OBJECTIVES: To analyze the changes in the proportion of blood units discarded from 1991 through 2001 at the Pro-Blood Foundation/Blood Center of Sao Paulo (Fundacao Pro-Sangue/Hemocentro de Sao Paulo), which is the largest blood bank in Latin America, and to determine the prevalence of infectious diseases among donors at the Blood Center in November 2001. METHODS: We compiled data concerning the discarding of blood units due to the presence of serological markers for communicable diseases at the Blood Center during the period from 1991 through 2001. To determine the prevalence of infectious diseases, 9 942 screened samples were analyzed in November 2001; all reactive samples underwent confirmatory tests. RESULTS: Over the study period there was a significant decrease in the percentage of units discarded, from 20% in 1991 to 9% in 2001. In November 2001 the prevalence of infectious diseases among donors was: 0.04% for human immunodeficiency virus (HIV), 0.21% for hepatitis C virus (HCV), 0.06% for human T-lymphotropic virus (HTLV), 0.14% for Chagas' disease, and 1.10% for syphilis. For hepatitis B virus, the prevalences found were: 0.14% for anti-HBc and HBsAg, 1.68% for anti-HBc and anti-HBs, and 1.67% for isolated anti-HBc. CONCLUSIONS: The decrease in the discarding of blood units and in infectious diseases among donors at the Blood Center of Sao Paulo reflects the increase in the Center's percentage of repeat donors. PMID- 12744787 TI - [Seropositivity for human T-lymphotropic virus types I and II among donors at the Municipal Blood Bank of Caracas and associated risk factors]. AB - OBJECTIVE: To conduct research at the Municipal Blood Bank of Caracas (MBBC) and find out the proportion of blood units discarded for being seropositive for human T-lymphotropic virus (HTLV) types I and II, the prevalence of that infection among their donors, and the probable risk factors for that infection among those HTLV-positive donors. METHODS: ELISA serological testing was done with 23 413 donors seen at the MBBC between July 2000 and April 2001. Samples that were repeat reactive (RR) with the ELISA underwent supplementary Western blot (WB) testing. Donors who had a positive or indeterminate WB result were scheduled for counseling in order to carry out confirmatory testing using nucleic acid amplification (NAA), to collect data on their risk background, and to advise them concerning their HTLV status. RESULTS: Of the 23 413 MBBC donors, 48 of them (0.2%) had a donation that was RR. Of those 48, 25 of them (52.1%) were positive on the WB (23 for HTLV-I and 2 for HTLV-II), 2 of them (4.1%) were indeterminate on the WB, 14 of them (29.2%) were negative, and 7 (14.6%) could not be evaluated. Of the 27 donors scheduled for counseling, 16 of them actually attended (14 WB-positive for HTLV-I, 1 WB-positive for HTLV-II, and 1 indeterminate). All 16 of them were positive with the confirmatory NAA testing. When these 16 seropositive donors were compared with a control group of seronegative donors, no significant differences were found with regard to age, sex, type of donation, number of previous donations, history of transfusions, and sexual behavior. However, significant differences were found in two areas: the seropositive donors were more likely to have used non-intravenous drugs (P < 0.05), and the seropositive donors were much more likely to have had an extended breast-feeding period (more than 2 years) as a child (P < 0.001). To assess the probability of mother-to-child transmission, six of the mothers of seropositive donors who had had an extended breast-feeding period were tested, and all six of those mothers were also found to be seropositive. With the 16 seropositive donors who were counseled, the spouse or partner of 13 of them was also tested; only 1 of those 13 was positive, but the oldest son of that couple was also HTLV positive. CONCLUSIONS: Of the donated blood, 0.2% of the units were discarded for being positive for HTLV-I or HTLV-II, and the prevalence found among the donors was 0.11%. Sexual transmission between an HTLV-positive donor and a partner or spouse was less frequent than was mother-to-child transmission. At present in Venezuela, blood banks are not required to screen donations for HTLV. Given our results at the MBBC, we believe serious consideration should be given to implementing serological screening for HTLV I/II among blood donors throughout Venezuela. PMID- 12744788 TI - Hemophilia care in the state of Rio de Janeiro, Brazil. AB - In the developing countries of the world, few people with hemophilia receive adequate care. Nevertheless, Brazil has made significant advances in the treatment of hemophilia over the last decade. The provision of factor concentrates imported by the Government of Brazil is gradually increasing, and patients receive the concentrates for free. A national register was established as well as a coordinated program for comprehensive care. Of the 6 297 persons with hemophilia in Brazil who were registered as of January 2001, 689 of them (11.1%) were registered in the state of Rio de Janeiro. Of those 689, 664 of them were being monitored at the state's coordinating blood transfusion center, which is located in the city of Rio de Janeiro. Among those 664, factor VIII inhibitors were identified in 81 of them (12.2%). Among 653 of the Rio de Janeiro patients who were tested for transfusion-transmitted diseases, the overall prevalence found was 41.5%, with the specific rates being 13.3% for human immunodeficiency virus (HIV), 2.9% for hepatitis B virus (HBV), and 39.4% for hepatitis C virus (HCV). The state of Rio de Janeiro has adopted a comprehensive hemophilia management approach that includes medical, psychological, and social care. As a result, the quality of life of hemophilia patients has improved noticeably. For example, the rate of hospitalization among patients fell by 30% between 1998 and 2001, and there has also been a decline in the school and work activities that they have missed. PMID- 12744789 TI - [External evaluation of serology results in blood banks in Colombia]. AB - OBJECTIVE: To analyze the serological results found in Colombian blood banks that participate in the external quality program (EQP) of that country's National Institute of Health, in order to improve the quality of the screening of blood for the main serological markers of transfusion-transmitted infectious diseases. METHODS: Each blood bank received a panel of six sera with different reactivity and positivity to hepatitis B surface antigen (HBsAg), as well as to antibodies to HIV 1-2, Trypanosoma cruzi (the causative agent of Chagas' disease), Treponema pallidum (the causative agent of syphilis), hepatitis B core (HBc) antigen, hepatitis C virus (HCV), and human T-lymphotropic virus (HTLV). The screening techniques used were enzyme-linked immunosorbent assay (ELISA), microparticle enzyme immunoassay (MEIA), and hemagglutination. With the panel sera, the participating blood banks were asked to apply the same tests that they use on a daily basis to screen blood units and to send their results to the National Blood Banks Unit of the Colombian National Institute of Health. RESULTS: Of the 46 blood banks participating in the EQP, 43 of them (93%) returned their results within the requested timeframe. The ELISA test was the one that was used most often (83.0%). There were a total of 49 (5%) false positive results and 12 (3%) false negative results. Of those 12 false negative results, 6 of them corresponded to the detection of syphilis, 2 to Chagas' disease, 2 to anti-HBc antibodies, 1 to anti-HCV antibodies, and 1 to HBsAg. Eighty percent of the discordant results came from 23 blood banks that each collected fewer than 6 000 units of blood per year, and 15% came from 5 blood banks that collected 6 000 to 12 000 units per year. One of the blood banks that collected more than 12 000 units annually had three false positive results, and none of those larger blood banks had any false positive results. CONCLUSIONS: The percentage of false negative results (3%) found during the EQP can be considered high, since tests that are negative during blood screening are not repeated, and the decision to declare a unit of blood suitable for transfusion is based on that single result. There is a need to thoroughly review the procedures for screening blood in Colombia, particularly at the centers that performed poorly in this EQP exercise. PMID- 12744790 TI - Trends in the profile of blood donors at a large blood center in the city of Sao Paulo, Brazil. AB - OBJECTIVE: To describe the trends in the profile of blood donors from 1995 through 2001 at a large blood center in the city of Sao Paulo, Brazil, particularly following the initiation in 1998 of marketing strategies aimed at substituting replacement donors with altruistic repeat donors. METHODS: Using an information system that had been established at the Pro-Blood Foundation/Blood Center of Sao Paulo (Fundacao Pro-Sangue/Hemocentro de Sao Paulo) in 1994, we collected information on sex, age, and type of donation for the years 1995-2001. We classified blood donors as either replacement blood donors (if they stated that the reason for donating was that they had a friend or relative in the hospital) or as altruistic donors. First-time blood donors were those who had not donated in our institution since the establishment of the information system. RESULTS: The percentage of repeat altruistic blood donors increased over time as first-time replacement donors declined for both genders. The proportion of altruistic donors climbed from 20% of all blood donors in 1995 to 57% in 2001. In 2001, first-time blood donors represented only 52% of all donors, as contrasted to 88% in 1995. Female donors increased from 20% to 37% of the donors over the period studied. CONCLUSIONS: Our data provide evidence that the Sao Paulo population has responded well to the marketing strategies that have been introduced in our institution. We believe that similar promotional efforts elsewhere in Brazil would produce comparable, positive results. PMID- 12744791 TI - [External performance evaluation of screening in blood banks in Argentina: results and strategies for improvement]. AB - OBJECTIVES: Because there was no program for the external evaluation of the quality of the screening for transfusion-transmitted infections (TTIs) in blood banks in Argentina, in 1999 the Hemotherapy Service of Garrahan Hospital, in Buenos Aires, launched an ongoing external evaluation program, with the support of the Pan American Health Organization and the Argentine Hemotherapy and Immunohematology Association. METHODS: A panel of 12 samples that were reactive to all the markers screened for in Argentine blood banks was distributed. The panel was delivered to 52 laboratories in 1999, 102 laboratories in 2000, and 118 laboratories in 2001. The participating laboratories were classified into one of four categories according to their performance: A: 0 or 1 false positive (FP) results; B: 2 or 3 FPs; C: 4 or more FPs; and D: at least 1 false negative result. Workshops were held with the participants in order to jointly analyze the results. RESULTS: Out of the laboratories that received the panel of samples, the percentage of laboratories that sent in their results was 92.3% in 1999, 92.2% in 2000, and 83.9% in 2001. These response levels demonstrate the interest in evaluation activities. However, the annual average percentage of the laboratories that received a "D" classification was very high (30%), which indicates that there are problems in the performance of TTI screening. This poor showing could be related to the difficulty in taking corrective measures throughout the system, given the large number of laboratories involved. CONCLUSIONS: Evaluations of this type should be included in Argentina's National Blood Program as a tool for the ongoing assessment of the blood banking situation, for decision-making, and for monitoring the quality of screening. PMID- 12744792 TI - [Neonatal screening for hemoglobinopathies in Rio de Janeiro, Brazil]. AB - OBJECTIVE: To describe the main results obtained in the first 15 months of neonatal screening for sickle cell disease in the state of Rio de Janeiro, Brazil, from August 2000 to November 2001. METHODS: Starting in August 2000, blood samples began to be collected for sickle cell disease screening from all newborns receiving care in primary health care clinics in the state of Rio de Janeiro. The samples were submitted to high-resolution liquid chromatography. If the resulting chromatogram was compatible with sickle cell disease, the child and the parents were referred for diagnostic confirmation and treatment. RESULTS: Between August 2000 and November 2001, 99 260 newborns were screened. There was one case of homozygous Hb C. On average, one of every 27 newborns who were screened presented sickle cell trait (Hb AS). Sickle cell disease was observed in 83 cases, or one new case in each 1 196 births. The 83 consisted of: 62 Hb S, 18 Hb SC, and 3 Hb SD. One child did not appear for diagnostic confirmation. The 82 children who were followed up by the program presented 15 intercurrent illnesses (upper respiratory infections, fever, splenic sequestration crises, hand-foot syndrome, and vascular occlusion), resulting in seven hospital admissions. Blood transfusions were necessary with 15 children, but none developed alloimmunization. All the other babies were doing well with the use of prophylactic penicillin. CONCLUSIONS: Our data show the importance of early diagnosis for sickle cell disease, so as to prevent the frequent infectious complications faced by these patients. PMID- 12744794 TI - [Quality control of immunologic testing of blood in the Region of the Americas]. AB - Assuring the best quality in transfusion medicine necessarily involves exercising exhaustive control over the entire system and developing protocols for procedures and techniques, the reagents and equipment used, personnel training, and so on. Additional guarantees can come from adequately organizing the work and perhaps automating processes in order to avoid errors. Another way to avoid errors in the laboratory is to register and continuously analyze the mistakes that do occur, since that makes it necessary to change procedures in order to avoid repeating them. External quality control programs (EQCP) make it possible to perform a periodic overall assessment of the suitability of techniques, reagents, and training of personnel in relation to the validity of the results obtained. By voluntarily participating in an EQCP, the blood center shows its commitment to the quality of care. In the Region of the Americas, the Pan American Health Organization's Immunohematology Quality Control Program has made it possible to objectively quantify the improvement attained in the formative level reached by participating blood banks in the four years the Program has been in operation. PMID- 12744793 TI - Evaluation of the concomitant use of two different EIA tests for HIV screening in blood banks. AB - OBJECTIVE: In 1998, the Brazilian Ministry of Health made it mandatory for all blood banks in the country to screen donated blood for human immunodeficiency virus (HIV) concomitantly using two different enzyme immunoassay (EIA) tests. Concerned with the best use of available resources, our objective with this study was to evaluate the usefulness of conducting two EIA screening tests instead of just one. METHODS: We analyzed data from 1999 through 2001 obtained by testing 698 191 units of donated blood using two EIA HIV screening tests concomitantly at the Pro-Blood Foundation/Blood Center of Sao Paulo (Fundacao Pro Sangue/Hemocentro de Sao Paulo), which is a major blood center in the city of Sao Paulo, Brazil. All samples reactive in at least one of the two EIA tests were submitted for confirmation by a Western blot (WB) test, and the persons who had donated those samples were also asked to return and provide a follow-up sample. RESULTS: Out of the 698 191 blood units that were donated, 2 718 of them (0.4%) had to be discarded because they were reactive to at least one of the EIA tests. There were two WB-positive donation samples that were reactive in only one HIV EIA screening test. On their follow-up samples, both donors tested WB-negative. These cases were considered false positive results at screening. Of the 2 718 donors who were asked to return and provide a follow-up sample, 1 576 of them (58%) did so. From these 1 576 persons, we found that there were two individuals who had been reactive to only one of the two EIA screening tests and who had also been negative on the WB at screening but who were fully seroconverted on the follow-up sample. We thus estimated that, in comparison to the use of a single EIA screening test, the use of two EIA screening tests would detect only one extra sample out of 410 700 units of blood. CONCLUSIONS: Our data do not support the use of two different, concomitant EIA screening tests for HIV. The great majority of HIV-positive donors have already seroconverted and will be detected using only a single EIA screening test. Only persons who are going through the process of seroconversion may be missed by using a single EIA test and detected using two EIAs for screening. To screen out these individuals and decrease the residual risk of HIV transmission from the blood donated in our center, the use of other techniques, such as nucleic acid testing (NAT) or a p24 antigen assay, would be more effective. PMID- 12744795 TI - [Guidelines for the transfusion of blood and its components]. AB - There is no general consensus on the appropriate use of blood and its components. The resulting lack of transfusion guidelines is a weakness in our hospitals and health centers that directly affects good transfusion practice. Based on a literature review, this piece offers some general guidelines on the appropriate use of transfusion therapy. These guidelines can serve as the basis for preparing more-specific guides that take into account the individual needs of each hospital center. PMID- 12744796 TI - Characteristics of HIV-infected childbearing women in Barbados. AB - OBJECTIVE: To describe the demographic profile, social and family characteristics, and lifestyle traits of HIV-infected childbearing women in the Caribbean nation of Barbados in comparison to a control group of HIV-negative women. METHODS: Data for this report were drawn from the Pediatrics HIV Surveillance Program of the Queen Elizabeth Hospital in Barbados. The data covered all HIV-infected women in the country who delivered between 1986-2000, with similar data coming from a control group of HIV-negative childbearing women. Routine information recorded during antenatal care was obtained from the women's case records. Additional data were collected from interviews with the women. RESULTS: There were 182 HIV-infected women who delivered during the study period, and a group of 202 childbearing women served as controls. In comparison to the control group, the HIV-infected women were younger, more often multiparous, and more likely to have been unemployed at the time of their pregnancy. The HIV infected women also had had an earlier onset of sexual activity, had had more sexual partners during their lifetime, and were more likely to be involved with an older sexual partner. At the time of giving birth most of the HIV-infected women were asymptomatic for AIDS and were living with either their parents (mother or father or both) or the baby's father. In addition, at the time of their six-weeks-postnatal visit, the large majority of the HIV-infected women were involved in caring for their children. The proportion of HIV-infected women who were diagnosed prior to childbirth increased significantly over the study period, rising from 25% during 1986-1990 to 82% during 1996-2000. Slightly over one-fifth of the HIV-infected women had had one or more subsequent pregnancies after they had learned that they were infected. CONCLUSIONS: The early age of sexual activity as well as repeated pregnancies, especially from different and older partners, may have contributed significantly to both vertical and horizontal HIV transmission in Barbados. Future studies of HIV incidence and its trend among childbearing women could be important for monitoring the HIV epidemic in this country. Many of the HIV-infected childbearing women in our study were unemployed, sick, and had multiple children. Therefore, to help them to plan for and cope with the disease and also the care of their children beyond the perinatal period, there is a need to provide the women with repeated counseling with continued follow-up and, where necessary, additional economic, social, and medical support. PMID- 12744797 TI - [Selective vs. mass treatment with antihelminthic drugs: experience in two hyperendemic communities]. AB - OBJECTIVE: To compare the prevalence and intensity of soil-transmitted nematode infection among children from two hyperendemic communities who were treated with antihelminthic drugs under two different regimens: selective or individual treatment was administered to members of one of the communities, and repeated mass treatment was given to the control community. METHODS: The study population was comprised of 909 children of both sexes between the ages of 2 and 13 from two peri-urban communities: Las Lomas and El Abastos, both in Santa Fe, Argentina. A prospective, longitudinal, quasi-experimental study was carried out in these communities. During the 22-month study period, 5 parasitologic controls were performed and results were qualitatively and quantitatively examined. Treatment with antihelminthic drugs was given at controls 0, 2, 3, and 4. At Las Lomas, selective treatment was administered to those in sample A (n = 55) who were positive for parasites; in El Abasto, mass treatment was given to all children, including those in sample B (n = 50). Both samples received follow-up. RESULTS: The prevalence and intensity of A. lumbricoides infections were found to be significant in El Abasto. No differences were detected in the case of T. trichiura. When prevalence and parasitic burden of A. lumbricoides were compared among controls within a single community (at the beginning and end of the study), no significant differences were observed in sample A, but a significant difference was seen in sample B. Insofar as T. trichiura is concerned, there were significant differences among controls in both samples. CONCLUSIONS: Only repeated and massive treatment significantly lowered the prevalence and intensity of A. lumbricoides infections during the study period. PMID- 12744798 TI - [Blood lead levels in children and environmental legacy of a lead foundry in Brazil]. AB - OBJECTIVE: To determine the blood lead levels in children living near an inactive lead foundry in the city of Santo Amaro da Purificacao, state of Bahia, in September of 1998; and to identify factors associated with differences in these levels. METHODS: Cross-sectional study with children between 1 and 4 years of age living within 1 km of the lead foundry. Mothers or guardians of 47 children answered a questionnaire concerning ingestion of clay, soil, plaster and/or other materials (pica), and other relevant epidemiological aspects. The concentration of lead in blood was determined by atomic absorption espectrophotometry. RESULTS: The mean blood lead level was 17.1 +/- 7.3 micro g/dL. Blood lead levels were approximately 5 micro g/dl greater among children with pica, regardless of age, visible presence of scum surrounding the home, employment status of the father, family history of lead poisoning, and malnutrition. CONCLUSIONS: The environmental legacy of the lead foundry, which was shut down in 1993, continues to represent a relevant risk factor for increased blood lead levels in children, especially those presenting pica. PMID- 12744799 TI - Prevalence of overweight and obesity among Costa Rican elementary school children. AB - OBJECTIVE: Given that excessive body weight during childhood influences the development of several chronic diseases in adulthood, this study was conducted to determine the prevalence of overweight and obesity in urban and rural Costa Rican elementary school children. METHODS: The study was carried out from July 2000 to April 2001. A total of 1 718 students ages 7-12 were selected from 34 schools in the capital city of San Jose and in other nearby urban and rural areas. Both younger children (ones aged 7 through 9 years) and older children (ones aged 10 through 12 years) with a body mass index (BMI) at or above the sex-specific 85th percentile were considered overweight. The younger children were classified as being obese if their triceps skinfold was greater than or equal to the 85th percentile for age and sex using the percentiles by age for children in the United States of America as normative standards. The older children were considered obese if they had a BMI at or above the sex-specific 85th percentile and both the triceps and subscapular skinfold thickness at or above the 90th percentile. RESULTS: The prevalence of overweight was 34.5%. Children aged 7-9, boys, children from urban areas, and children of a higher socioeconomic status had a higher prevalence of overweight. The prevalence of obesity was 26.2%. A higher prevalence of obesity was found among children aged 7-9, boys, children from urban areas, and children of middle socioeconomic status. CONCLUSIONS: Given the high prevalence of obesity that we found in the Costa Rican children, primary and secondary prevention measures are needed in order to reduce the proportion of deaths due to chronic nontransmissible diseases among Costa Rican adults in the coming decades. PMID- 12744800 TI - [Helmet use by motorcyclists injured in traffic accidents in Londrina, southern Brazil]. AB - OBJECTIVE: To describe helmet use among motorcyclists injured in traffic accidents in Londrina, a medium-sized city in Parana, a state in southern Brazil, and to identify factors associated with not wearing a helmet at the time of the accident. METHODS: We analyzed data concerning motorcycle users (drivers and passengers) who received care in 1998 from the only pre-hospitalization trauma and emergency care ambulance service in Londrina. The following variables were assessed: helmet use at the time of the accident; age; sex; seating position (driver or passenger); smell of alcohol on the breath; time of the day, day of the week, and month when the accident happened; and location of the accident (in the downtown area or outside of it). RESULTS: The average rate of helmet use was 63.2%. Factors independently associated with not using a helmet were (according to strength of association): being younger than 18 years of age (odds ratio (OR) = 6.61), having alcohol on the breath (OR = 3.93), accident occurring at night (OR = 2.51), accident happening outside the city's downtown area (OR = 2.27), and accident taking placing during the weekend (OR = 2.25). CONCLUSIONS: It is imperative to implement public policies aimed at promoting safety for motorcyclists, ongoing education in defensive driving, and the use of helmets and other safety equipment. Interventions should take into account that driving behaviors depend not just on individuals but also on the overall social and historical context. PMID- 12744803 TI - [The relationships between organizations for technical cooperation in health and private consulting businesses]. AB - In recent years, agencies that provide technical cooperation in health have increased their contractual relationships with private consulting entities. This has made it possible to respond in a timely manner to the support needs that countries have, to develop skills at the national level, and to reduce the operating costs for the cooperation agencies. However, these relationships risk moving the cooperation agencies away from generating ideas and new knowledge, which, until recently, was considered one of their essential roles. Contracting with private enterprises will almost certainly increase in the coming years. This makes it worth reviewing the tasks that correspond to the cooperation agencies in this scenario as well as mechanisms to see that these relationships result in the greatest benefit for deprived groups. Actions that can be undertaken immediately include organizing the "structural capital" (such as programs, databases, strategies, and organizational "culture," structure, systems, and procedures) of the technical cooperation agencies, precisely identifying tasks that cannot be delegated, and adequately designing and controlling terms of reference. PMID- 12744804 TI - [Biotechnologies to advance health in developing countries]. PMID- 12744806 TI - Collection, preservation, and transport of field specimens for molecular investigations. PMID- 12744807 TI - Impaired clearance of apoptotic cells in systemic lupus erythematosus: challenge of T and B cell tolerance. PMID- 12744808 TI - Rationale for the use of cyclooxygenase-2-specific nonsteroidal anti-inflammatory drugs in ankylosing spondylitis: the available evidence. PMID- 12744809 TI - Rheumatic manifestations of diabetes mellitus. AB - An epidemic of diabetes is currently raging in the US and developed countries, and is affecting almost 10% of the US population. The association between several rheumatic disorders and diabetes mellitus is gaining attention, and with recent data showing that more than 30% of patients with type 1 or type 2 diabetes have some hand or shoulder diseases, the magnitude of this problem is becoming more evident. Although some syndromes are observed exclusively in patients with diabetes, the majority of the rheumatic diseases found in patients with diabetes are also seen in the non-diabetes population, albeit at a much lower prevalence. The exact mechanisms by which the specific metabolic abnormalities of diabetes impact on the pathogenesis of its rheumatic manifestations are not clear. Further studies are needed to better understand the role of diabetes on the development of these disorders. PMID- 12744810 TI - Behcet's syndrome: an update. AB - The pathogenic mechanisms in Behcet's syndrome are largely unknown. An autoantigen role for human leukocyte antigen B51 has been proposed. The reasons behind the thrombophilia are also not clear. Endothelial pathology could be the main culprit. The recently proposed association between familial Mediterranean fever and Behcet's syndrome might not be well founded. The long-term prognosis is more guarded among the young and among males. However, the disease burns out in many cases. Clinicians are getting better at management, and have better understanding of the old drugs, such as colchicine, and have new and potent drugs like tumor necrosis factor-alpha inhibitors. PMID- 12744811 TI - Rheumatic manifestations of hepatitis C infection. AB - Among chronic viral infections, hepatitis C virus (HCV) infection is uniquely associated with an array of rheumatic manifestations and autoimmune laboratory findings. These include, among others, arthralgias, arthritis, fatigue, fibromyalgia, vasculitis, and sialadenitis (Sjogren-like). The mechanisms that are involved in the pathogenesis of these diverse manifestations have not yet been clarified. Regardless of the direct or indirect pathogenetic role of HCV in these clinical entities, the concomitant presence of a chronic viral infection creates a number of diagnostic and therapeutic problems. This is particularly true when immunosuppressive therapy is needed for control of disease activity (eg, HCV-associated cryoglobulinemic vasculitis). The emerging treatment options for chronic HCV offer a major chance for viral eradication and conceivably for cure of these HCV-associated conditions. In this review, the recent advances in the epidemiology, pathogenesis, clinical findings, and treatment of HCV associated rheumatic conditions are presented. PMID- 12744812 TI - Rheumatic manifestations of human immunodeficiency virus. AB - Since July 1983, various rheumatic, musculoskeletal, or other immune disorders characterized by dysregulation have been associated with HIV and AIDS. Infections occur, but with a lower frequency than expected in a patient population with a disorder primarily characterized by significant cellular immune deficiencies. Reactive arthritis, psoriatic arthritis, acute nonspecific arthritis, Sjogren syndrome, and inflammatory myositis have been reported. The initial reports of "acute painful joints," however, have not maintained prominence in the literature. A review of the literature over the past several years has reinforced the perception that the initial excitement over a possible association between HIV and AIDS and rheumatic or definable autoimmune disorders remains limited to a small segment of illnesses dominated by the reactive arthritidies. PMID- 12744813 TI - The case for uricase in gout. PMID- 12744814 TI - Hydroxyapatite deposition disease of the joint. AB - Basic calcium phosphate (BCP) crystals include partially carbonate-substituted hydroxyapatite, octacalcium phosphate, and tricalcium phosphate. They may form deposits, which are frequently asymptomatic but may give rise to a number of clinical syndromes including calcific periarthritis, Milwaukee shoulder syndrome, and osteoarthritis, in and around joints. Recent data suggest that magnesium whitlockite, another form of BCP, may play a pathologic role in arthritis. Data from the past year have provided further understanding of the mechanisms by which BCP crystals induce inflammation and degeneration. There remains no specific treatment to modify the effects of BCP crystals. Although potential drugs are being identified as the complex pathophysiology of BCP crystals is unraveled, much work remains to be done in order to translate research advances to date into tangible clinical benefits. PMID- 12744816 TI - Hyperuricemia and gout. AB - Gout is not a new disease for clinicians; nevertheless, there are still many secrets awaiting discovery for improving knowledge with respect to uric acid metabolism and monosodium urate crystal-induced inflammation. This review of the literature will focus on new insights on the pathogenesis of idiopathic hyperuricemia, and on secondary hyperuricemia and gout. There are also important advances on the pathophysiology of acute gout, especially as a self-limited process (switch from monocyte to macrophage, peroxisome proliferator activated receptor-gamma, and nitric oxide), but also of chronic gouty arthropathy. Armaments for treating hyperuricemia and gout may be already improved by losartan or fenofibrate and, in the future, by urate oxydase-polyethylene glycol 20 and renal handling regulatory molecules. Finally, control of hyperuricemia may also be considered in the prevention and treatment of cardiovascular disease. PMID- 12744815 TI - Matrix vesicles and calcification. AB - Matrix vesicles (MVs) are extracellular, 100 nM in diameter, membrane-invested particles selectively located at sites of initial calcification in cartilage, bone, and predentin. The first crystals of apatitic bone mineral are formed within MVs close to the inner surfaces of their investing membranes. Matrix vesicle biogenesis occurs by polarized budding and pinching-off of vesicles from specific regions of the outer plasma membranes of differentiating growth plate chondrocytes, osteoblasts, and odontoblasts. Polarized release of MVs into selected areas of developing matrix determines the nonrandom distribution of calcification. Initiation of the first mineral crystals, within MVs (phase 1), is augmented by the activity of MV phosphatases (eg, alkaline phosphatase, adenosine triphosphatase and pyrophosphatase) plus calcium-binding molecules (eg, annexin I and phosphatidyl serine), all of which are concentrated in or near the MV membrane. Phase 2 of biologic mineralization begins with crystal release through the MV membrane, exposing preformed hydroxyapatite crystals to the extracellular fluid. The extracellular fluid normally contains sufficient Ca2+ and PO4(3-) to support continuous crystal proliferation, with preformed crystals serving as nuclei (templates) for the formation of new crystals by a process of homologous nucleation. In diseases such as osteoarthritis, crystal deposition arthritis, and atherosclerosis, MVs initiate pathologic calcification, which, in turn, augments disease progression. PMID- 12744818 TI - Crystal deposition disease of the shoulder (including calcific tendonitis and milwaukee shoulder syndrome). AB - Calcific tendinitis of the shoulder is a dynamic process. Osteopontin is present in cells surrounding tendon calcifications. Resorption is probably mediated by cathepsin K-containing multinucleated giant cells. Rotator cuff tears are associated with an inflammatory response based on the presence of interleukin-1 and a proliferative synovitis. Metalloproteinases are found in the synovial fluids of patients with rotator cuff tears. Some patients with large rotator cuff tears progress to a severe destructive arthropathy characterized by large joint effusions, which are noninflammatory but contain basic calcium phosphate crystals. These crystals stimulate metalloproteinase production in vitro and also suppress metalloproteinase inhibitor production. Mutations in the ank gene result in decreased extracellular inorganic pyrophosphate in murine progressive ankylosis, and increased extracellular inorganic pyrophosphate in some cases of familial chondrocalcinosis. PMID- 12744817 TI - Calcium pyrophosphate dihydrate and hydroxyapatite crystal deposition in the joint: new developments relevant to the clinician. AB - The major types of crystals containing calcium, which causes arthropathy and periarticular disease, are calcium pyrophosphate dihydrate and basic calcium phosphates, including hydroxyapatite. Exciting advances include the identification of mutations in the gene ANKH associated with disordered inorganic pyrophosphate (PPi) transport in some kindred with familial chondrocalcinosis linked to chromosome 5p. In addition, central basic mechanisms governing cartilage calcification and their relationship to aging and osteoarthritis have now been elucidated. These include the role of plasma cell glycoprotein-1, the PPi-generating ecto-enzyme, in chondrocalcinosis and the linkage of low- grade inflammation to expression and activation of two cartilage-expressed transglutaminase isoenzymes with direct calcification-stimulating activity. This review discusses clinically pertinent new information on pathogenesis. The authors also address, in detail, current diagnostic and therapeutic issues pertaining to calcium pyrophosphate dihydrate and hydroxyapatite crystal deposition in the joint, as well as possible therapeutic directions for the future. PMID- 12744820 TI - Treatment of Extraintestinal Manifestations in Inflammatory Bowel Disease. AB - Extraintestinal manifestations (EIM) of inflammatory bowel disease (IBD) occur rather frequently and may be found in up to 30% of patients. However, surprisingly few randomized, controlled studies have been conducted that were specifically aimed at the treatment of EIM of IBD patients. Therefore, most therapies of EIM are empiric or deduced from studies in populations with other type of patients. EIM may be associated with active IBD. Treatment of active IBD is, therefore, the mainstay of treatment of EIM. Lifestyle modification as a means of therapy is a recent subject of study in chronic conditions, such as IBD. Based on epidemiologic and experimental findings, EIM of various tracts can be modified by optimizing alimentary intake, refraining from sedentary lifestyle, and adapting smoking habits. Not many new drugs for treatment of EIM have been developed during the past few years; the role of infliximab has been extended in particular in Crohn's disease-related EIM. Careful consideration of prescribed drugs remains necessary due to potential interaction with the course of IBD. PMID- 12744819 TI - Treatment of Crohn's Disease of Inflammatory, Stenotic, and Fistulizing Phenotypes. AB - The heterogeneous nature of Crohn's disease (CD) is reflected in the diversity of treatment options available for individual patients. The stratification of CD patients into more homogeneous groups based on disease location and disease behavior may provide clinicians with a more focused approach to therapeutic decision-making. Uncomplicated disease behaviors are typically treated medically. When complications arise and patterns of disease become more aggressive, combined medical and surgical approaches are often necessary and yield favorable results. The surgical management of CD can be as complex as the disease itself, and should involve a surgeon who professes a special expertise in inflammatory bowel disease. Progress in our understanding of the role of the interaction between the environment and the immune system in disease development has led to major advancements in the area of CD therapeutics. Current therapies target the various elements of the inflammatory cascade implicated in the pathogenesis of CD. The anti-inflammatory properties of the pharmacologic therapies presented in this review vary from actions that are extremely broad to those that are cellular or cytokine specific. Maximizing the efficacy of CD-directed therapies while minimizing their toxicity remains the principal objective in developing management strategies for CD patients. Maintaining good quality of life and maximizing adherence to therapies are also important considerations. Despite the various therapeutic options available for CD patients, chosen therapies should be based on the overall treatment goal for individual patients. Therapeutics can be broadly categorized as induction therapies (goal to treat active disease) and maintenance therapies (goal to prevent relapse of disease). PMID- 12744821 TI - Inflammatory Bowel Disease After Age 60. AB - New-onset idiopathic inflammatory bowel disease (IBD) is not uncommon among the elderly, although more common are colonic infection, ischemia, or neoplasia, all of which may mimic IBD. Although the clinical presentation of IBD in the elderly often resembles that of younger subjects, atypical manifestations are common and may lead to difficulty in diagnosis. Much progress has been made in both medical and surgical therapy for IBD, but such therapy poses additional challenges in the elderly, who are more likely to experience adverse effects of medications or complications of surgery. The elderly generally have a favorable outcome to both medical and surgical therapy for IBD. Although concern about possible untoward effects of therapy is warranted, treatment should not be withheld because of fear of complications. PMID- 12744822 TI - Inflammatory Bowel Disease During Pregnancy. AB - Physicians treating patients with Crohn's disease and ulcerative colitis will often need to care for them throughout pregnancy and deal with the surrounding issues of fertility, childbirth, and sexuality. Patients often worry about continuing medications during pregnancy and feel particularly at risk for poor birth outcomes. However, because pregnancy outcomes are most closely tied to disease activity at the time of conception, patients who are in remission when they conceive will have the most successful pregnancies. The overriding principle in treating pregnant patients with inflammatory bowel disease (IBD) is continued and close surveillance of disease activity, with aggressive medical, and if indicated, surgical treatment. With few exceptions, medicines used to induce remission before pregnancy should be continued throughout pregnancy. Pregnant women with active IBD should be followed by a gastroenterologist with experience in the issues surrounding pregnancy, and by an obstetrician with access to a tertiary referral center. Properly treated and followed, patients with IBD can expect outcomes from their pregnancies that approximate those of patients without the disease. PMID- 12744823 TI - Endoscopic Therapy for Inflammatory Bowel Disease. AB - Endoscopic therapy can be employed and may be useful in inflammatory bowel disease patients with dysplastic polyps, inflammatory strictures (enteric and biliary), bleeding, and for treatment of some complications of continent ileostomies. Dysplastic polyps can be removed endoscopically safely and effectively without resorting to colectomy, as long as there is no other detectable dysplasia in flat mucosa and complete removal can be assured (by biopsy of adjacent mucosa and close follow-up). Some colonic strictures in Crohn's disease can be dilated using endoscopes of graded caliber or with through the-scope balloons, with or without stent placement. Endoscopy is most useful in dilating anastomotic strictures and less useful for long strictures in active inflammatory disease. Strictures in ulcerative colitis are suspicious for neoplasia and, if dilated at all, should be biopsied extensively and followed closely. We prefer colectomy to endoscopic dilatation for strictures in ulcerative colitis. Biliary endoscopy can be used to dilate strictures in primary sclerosing cholangitis and to sample these areas for malignancy. Although dilatation may improve morbidity and prolong survival, it may not prevent progression to cirrhosis. In cholangiocarcinomas, endoscopic dilatation with or without stents can offer palliation and perhaps be used to deliver photodynamic therapy. Injection and sclerotherapy can be employed in Crohn's disease bleeding from a discrete site. In obstructed continent ileostomies (both Kock pouches and pelvic ileoanal reservoirs), endoscopy can be employed effectively to both determine the cause of the obstruction and re-establish patency. We do not advocate endoscopic treatment of toxic megacolon because of the heightened risk of perforation. Endoscopic therapy of Crohn's fistulas is a possible emerging technology, but it has not been used in large cohorts of patients. PMID- 12744824 TI - Hirschsprung's Disease. AB - Hirschsprung's disease (HSCR) is the most common congenital malformation of the enteric nervous system and requires early diagnosis and surgical repair for the best comprehensive outcome. The early diagnosis of this disorder permits the use of primary endorectal pull-through (PERPT), which is now the definitive surgical therapy for HSCR. PERPT has become the preferred method of treatment for HSCR, and large numbers of successfully treated patients have been described in the recent medical literature. The rate of postoperative complications is generally similar to that following a two-stage surgical repair, but PERPT patients may be at a slightly higher risk for Hirschsprung's-associated enterocolitis. Despite recent surgical advances in the treatment of HSCR, a two-stage surgical repair involving a temporary diverting colostomy may still be necessary in up to one third of patients. Candidates for a staged repair include those HSCR patients with long-segment or total colonic disease or when there has been a delay in diagnosis that results in a markedly dilated proximal colon or patient clinical instability. Internal anal sphincter hypertonicity, occurring either as isolated primary anal achalasia or as a postoperative complication, can be successfully managed by either botulinum toxin injections or anal myectomy. The measurement of colonic motility in surgically repaired patients with a long-standing postoperative abnormality of bowel function can identify several distinct motility disorders that are amenable to separate and individualized therapies. The single most important element in the management of HSCR remains the clinical judgement of the surgeon of record, who utilizes all discernible clinical data to elect the manner of surgical repair in a given patient. PMID- 12744825 TI - Chronic Anal Fissure. AB - Diagnosis of chronic anal fissure is easy and common in clinical practice. Little is known about the etiology and pathogenesis of this disorder. Current investigations consider anal sphincteric hypertonia and ischemia as primary factors in the appearance and maintenance of this lesion. Recurrence rate after healing is high, so anal fissure may be a chronic disease that evolves depending on sphincteric features. Conservative measures to avoid constipation, including fiber intake, are useful to improve symptomatology, achieve healing, and reduce recurrence. Surgical treatment is the most effective procedure for chronic anal fissure. Lateral internal sphincterotomy achieves healing in most cases (more than 95%) and the recurrence rate is low (1% to 3%). However, permanent fecal incontinence may appear after surgery and available data about this complication are controversial. In recent years, chemical sphincterotomy has been developed as an option in the treatment of chronic anal fissure. This medical option aims to achieve the effectiveness of surgery without side effects, by means of a temporary decrease of anal pressures that allows fissures to heal. Local injection of botulinum toxin into the anal sphincter is the most successful medical option, nearly as effective as surgery and without significant adverse effects (transitory episodes of mild fecal incontinence). Although more studies are needed to establish the method of administering this treatment, in our opinion botulinum toxin is an effective option in a high percentage of cases, especially in patients who risk developing incontinence. Compared with botulinum toxin, topical nitroglycerine ointments, which produce a transitory sphincteric relaxation, have the advantage of being a simple and accessible procedure. However, we think that this option should not be a first choice because its effectiveness is lower compared with surgery (about 60% to 70%), its compliance with the application could be poor, and it has a greater percentage of side effects (eg, headache). Other topical treatments (eg, calcium channel antagonist or cholinergic agonists agents) appear to be as effective as nitroglycerine agents and do not have significant adverse effects, but little data exist about these options. In our opinion, treatment of chronic anal fissure must be individualized, depending on the clinical profile of patients. Medical treatment, especially injection of botulinum toxin, should be taken into account if risk for developing incontinence is suspected. PMID- 12744826 TI - Proctitis and Sexually Transmissible Diseases of the Colon. AB - In patients with gastrointestinal symptoms and a history of anal-receptive intercourse, a careful but explicit history concerning sexual preferences and practices is required. Clinically, four infectious syndromes can be distinguished: perianal disease, proctitis, proctocolitis, and enteritis. Each of these syndromes may be caused by a certain range of pathogens. In immunocompromised patients, additional diseases must be considered. Clinical examination, a rectal swab with incubation in specific transport media, as well as a Gram stain of mucus or pus from a rectal swab, in combination with a rectosigmoidoscopy are helpful in differentiating specific diseases. Stool cultures and serologic examinations may identify specific causes of colitis or enteritis, but are of minor importance in the case of anorectal disease. Good treatments are available for most sexually transmitted anorectal diseases. An additional role for the treating physician and other healthcare providers is to educate and counsel patients and their sex partners on ways to adopt safer sexual behavior. PMID- 12744827 TI - Longevity of granular iron in groundwater treatment processes: changes in solute transport properties over time. AB - Although progress has been made toward understanding the surface chemistry of granular iron and the mechanisms through which it attenuates groundwater contaminants, potential long-term changes in the solute transport properties of granular iron media have until now received relatively little attention. As part of column investigations of alterations in the reactivity of granular iron, studies using tritiated water (3H(2)O) as a conservative and non-partitioning tracer were periodically conducted to independently isolate transport-related effects on performance from those more directly related to surface reactivity. Hydraulic residence time distributions (HRTDs) within each of six 39-cm columns exposed to bicarbonate solutions were obtained over the course of 1100 days of operation. First moment analyses of the data revealed generally modest increases in mean pore water velocity (v) over time, indicative of decreasing water-filled porosity. Gravimetric measurements provided independent estimates of water-filled porosity that were initially consistent with those obtained from 3H(2)O tracer tests, although at later times, porosities derived from gravimetric measurements deviated from the tracer test results owing to mineral precipitation. The combination of gravimetric measurements and 3H(2)O tracer studies furnished estimates of precipitated mineral mass; depending on the assumed identity of the predominant mineral phase(s), the porosity decrease associated with solute precipitation amounted to 6-24% of the initial porosity. The accumulation of mineral and gas phases led to the formation of regions of immobile water and increased spreading of the tracer pulse. Application of a dual-region transport model to the 3H(2)O breakthrough curves revealed that the immobile water-filled region increased from initially negligible values to amounts ranging between 3% and 14% of the total porosity in later periods of operation. For the aged columns, mobile-immobile mass transfer coefficients (k(mt)) were generally in the range of 0.1-1.0 day(-1) and reflected a slow exchange of 3H(2)O between the two regions. Additional model calculations incorporating sorption and reaction suggest that although changes in HRTD can have an appreciable effect on trichloroethylene (TCE) transformation, the effect is likely to be minor relative to that stemming from passivation of the granular iron surface. PMID- 12744828 TI - The effect of local-scale physical heterogeneity and nonlinear, rate-limited sorption/desorption on contaminant transport in porous media. AB - Nonideal transport of contaminants in porous media has often been observed in laboratory characterization studies. It has long been recognized that multiple processes associated with both physical and chemical factors can contribute to this nonideal transport behavior. To fully understand system behavior, it is important to determine the relative contributions of these multiple factors when conducting contaminant transport and fate studies. In this study, the relative contribution of physical-heterogeneity-related processes versus those of nonlinear, rate-limited sorption/desorption to the observed nonideal transport of trichloroethene in an undisturbed aquifer core was determined through a series of miscible-displacement experiments. The results of experiments conducted using the undisturbed core, collected from a Superfund site in Tucson, AZ, were compared to those obtained from experiments conducted using the same aquifer material packed homogeneously. The results indicate that both physical and chemical factors, specifically preferential flow and associated rate-limited diffusive mass transfer and rate-limited sorption/desorption, respectively, contributed to the nonideal behavior observed for trichloroethene transport in the undisturbed core. A successful prediction of trichloroethene transport in the undisturbed core was made employing a mathematical model incorporating multiple sources of nonideal transport, using independently determined model parameters to account for the multiple factors contributing to the nonideal transport behavior. The simulation results indicate that local-scale physical heterogeneity controlled the nonideal transport behavior of trichloroethene in the undisturbed core, and that nonlinear, rate-limited sorption/desorption were of secondary importance. PMID- 12744829 TI - In situ biodegradation determined by carbon isotope fractionation of aromatic hydrocarbons in an anaerobic landfill leachate plume (Vejen, Denmark). AB - Concentrations and isotopic compositions (13C/12C) of aromatic hydrocarbons were determined in eight samples obtained from the strongly anoxic part of the leachate plume downgradient from the Vejen Landfill (Denmark), where methanogenic, sulfate-reducing and iron-reducing conditions were observed. Despite the heterogeneous distribution of the compounds in the plume, the isotope fractionation proved that ethylbenzene and m/p-xylene were subject to significant biodegradation within the strongly anoxic plume. The isotope fractionation factors (alphaC) for the degradation of the m/p-xylene (1.0015) and ethylbenzene (1.0021) obtained from the field observations were similar to factors previously determined for the anaerobic degradation of toluene and o-xylene in laboratory experiments, and suggest that in situ biodegradation is one major process controlling the fate of these contaminants in this aquifer. The isotope fractionation determined for 1,2,4-trimethylbenzene and 2-ethyltoluene suggested in situ biodegradation; however, the isotopic composition did not correlate well with the respective concentration as expressed by the Rayleigh equation. Some other compounds (1,2,3-trimethylbenzene, o-xylene, naphthalene and fenchone) did not show significant enrichments in delta13C values along the flow path. The compound concentrations were too low for accurate isotope analyses of benzene, toluene, 1- and 2-methylnaphthalene, while interferences in the chromatography made it impossible to evaluate the isotopic composition for 4-ethyltoluene, 1,3,5 trimethylbenzene and camphor. In addition to demonstrating the potential of assessing isotopic fractionation as a means for documenting the in situ biodegradation of complex mixtures of aromatic hydrocarbons in leachate plumes, this study also illustrates the difficulties for data interpretation in complex plumes and high analytical uncertainties for isotope analysis of organic compounds in low concentration ranges. PMID- 12744830 TI - Scaling DNAPL migration from the laboratory to the field. AB - A particular problem with the release of dense nonaqueous phase liquids (DNAPLs) into the environment is identifying where the DNAPL is and if it is still moving. This question is particularly important at sites where thousands of cubic meters of DNAPLs were disposed of. To date, results from laboratory models have not been scaled to predict analogous migration at the larger length and time scales appropriate for sites where large volumes of DNAPLs were released. Modified inspectional analysis is a technique for developing scaling relationships through nondimensionalizing the governing equations. It was applied in this study to scale observations of DNAPL migration in a laboratory model to four hypothetical scenarios in the field where large volumes of DNAPL were released. One scenario was compared to a large DNAPL spill site. The length and time scales of DNAPL movement predicted from our analysis are consistent with those predicted from a numerical model of this site. To our knowledge, this is the first application of modified inspectional analysis for release of DNAPLs in a laboratory model. This methodology may prove useful for scaling results from other laboratory investigations of DNAPL migration to field-scale systems. PMID- 12744831 TI - Flow behavior and residual saturation formation of liquid carbon tetrachloride in unsaturated heterogeneous porous media. AB - The formation of residual, discontinuous nonaqueous phase liquids (NAPLs) in the vadose zone is a process that is not well understood. To obtain data that can be used to study the development of a residual NAPL saturation in the vadose zone and to test current corresponding models, detailed transient experiments were conducted in intermediate-scale columns and flow cell. The column experiments were conducted to determine residual carbon tetrachloride (CCl(4)) saturations of two sands and to evaluate the effect of CCl(4) vapors on the water distribution. In the intermediate-scale flow cell experiment, a rectangular zone of the fine grained sand was packed in an otherwise medium-grained matrix. A limited amount of CCl(4) was injected from a small source and allowed to redistribute until a pseudo steady state situation had developed. A dual-energy gamma radiation system was used to determine fluid saturations at numerous locations. The experiments clearly demonstrated the formation of residual CCl(4) saturations in both sands. Simulations with an established multifluid flow simulator show the shortcomings of current relative permeability-saturation-capillary pressure (k-S-P) models. The results indicate that nonspreading behavior of NAPLs should be implemented in simulators to account for the formation of residual saturations. PMID- 12744832 TI - A reaction-transport model for calcite precipitation and evaluation of infiltration fluxes in unsaturated fractured rock. AB - The percolation flux in the unsaturated zone (UZ) is an important parameter addressed in site characterization and flow and transport modeling of the potential nuclear-waste repository at Yucca Mountain, NV, USA. The US Geological Survey (USGS) has documented hydrogenic calcite abundances in fractures and lithophysal cavities at Yucca Mountain to provide constraints on percolation fluxes in the UZ. The purpose of this study was to investigate the relationship between percolation flux and measured calcite abundances using reactive transport modeling. Our model considers the following essential factors affecting calcite precipitation: (1) infiltration, (2) the ambient geothermal gradient, (3) gaseous CO(2) diffusive transport and partitioning in liquid and gas phases, (4) fracture matrix interaction for water flow and chemical constituents, and (5) water-rock interaction. Over a bounding range of 2-20 mm/year infiltration rate, the simulated calcite distributions capture the trend in calcite abundances measured in a deep borehole (WT-24) by the USGS. The calcite is found predominantly in fractures in the welded tuffs, which is also captured by the model simulations. Simulations showed that from about 2 to 6 mm/year, the amount of calcite precipitated in the welded Topopah Spring tuff is sensitive to the infiltration rate. This dependence decreases at higher infiltration rates owing to a modification of the geothermal gradient from the increased percolation flux. The model also confirms the conceptual model for higher percolation fluxes in the fractures compared to the matrix in the welded units, and the significant contribution of Ca from water-rock interaction. This study indicates that reactive transport modeling of calcite deposition can yield important constraints on the unsaturated zone infiltration-percolation flux and provide useful insight into processes such as fracture-matrix interaction as well as conditions and parameters controlling calcite deposition. PMID- 12744833 TI - An improved collocation method for solving the Henry problem. AB - The original Henry problem is characterized with severe (albeit unphysical) sea boundary condition difficult to handle with numerical methods. In this paper, we present an improved numerical solution of the Henry problem. Presented numerical model solves the steady-state dimensionless equations by the collocation method named Fup Fragment Collocation Method (FFCM) and uses R(bf) basis functions of Fup(2) (x, y) type. This method enables application of the classical formulation and high approximation accuracy as proven in comparison with published solutions. Particular difficulty in solving the original Henry problem is in the accurate representation of the seepage face due to the fixed sea concentration at the sea boundary. The results of the original Henry problem formulation and problem with modified boundary conditions indicate the accuracy and robustness of the FFCM in describing the discharge area of the considered problem. PMID- 12744834 TI - Sound analysis in auditory cortex. AB - Not so long ago, the auditory cortex took a back seat to the visual system in neuroscience research. With some notable exceptions outside the primate order, such as the classic work on echo-locating bats, owls and birdsong, the auditory cortex has been overlooked: only a few investigators were involved in understanding the structure and function of the monkey auditory cortex, and even fewer had the means to study its human counterpart. This situation has undergone a dramatic change in the past decade or so. Spurred on by advances in primate neurophysiology and neuroanatomy, and especially by developments in functional neuroimaging, substantial progress is now being made into understanding how the human auditory cortical system works. PMID- 12744835 TI - Response: sound analysis in auditory cortex--from temporal decomposition to perception. PMID- 12744836 TI - Neurotrophins in myelination: a new role for a puzzling receptor. AB - Binding of neurotrophins to p75(NTR) was recently identified as a positive signal for myelination by Schwann cells. This new finding adds yet another distinct biological role to the growing list of functions of p75(NTR) in the nervous system and identifies a novel target for promoting remyelination in peripheral neuropathies or post-nerve-injury. PMID- 12744837 TI - D1 and NMDA receptors hook up: expanding on an emerging theme. AB - In the dominant paradigm for functional interactions between G-protein-coupled receptors (GPCRs) and ligand-gated ion channels, the mantra is that the interactions are indirect, indirect, indirect. It is commonly understood that activating GPCRs engages one or more sets of intracellular signaling cascades that indirectly affect the function or localization of a cohort of ligand-gated ion channels. Conversely, stimulating ligand-gated channels, particularly those that are permeable to Ca(2+), indirectly produces effects on GPCRs and/or on GPCR activated signaling cascades. The foundation of this dichotomous world-view shook (just a little) when it was discovered that D5 dopamine receptors bind directly to GABA(A) receptors, and that this direct interaction reciprocally affects the signaling of both types of receptor. Now, D1 dopamine receptors also have got into the act! PMID- 12744838 TI - Pursuit neurons encode 3D space: is the cortex nervous and tensor? AB - Fukushima and colleagues have found single neurons in monkey frontal cortex that control pursuit eye movements by representing extra-personal space in a 3D coordinate frame. These cells might link the 2D depiction of visual space by the retina with the need of the body to act in the 3D world. We could be on the verge of a new understanding of coordinate transformations serving sensory-motor integration in cerebral cortex. PMID- 12744839 TI - PerleCan fix your muscle AChEs. AB - Efficient and accurate synaptic transmission requires proper localization of numerous signaling proteins in the synaptic membrane. At the neuromuscular junction, the nicotinic ACh receptor mediates postsynaptic depolarization, and acetylcholinesterase (AChE) terminates this process by hydrolyzing ACh. The mechanism by which the nerve directs receptor localization is understood in considerable detail; AChE clustering, by contrast, has received much less attention. Now, in a recent paper in Nature Neuroscience, the laboratories of Yoshiko Yamada and Richard Rotundo report that AChE clustering at the postsynaptic membrane requires perlecan, which binds both AChE and dystroglycan. PMID- 12744840 TI - The fundamental role of memory in olfactory perception. AB - Current emphasis on odorant physiochemical features as the basis for perception largely ignores the synthetic and experience-dependent nature of olfaction. Olfaction is synthetic, as mammals have only limited ability to identify elements within even simple odor mixtures. Furthermore, olfaction is experience-bound, as exposure alone can significantly affect the extent to which stimuli can be discriminated. We propose that early analytical processing of odors is inaccessible at the behavioral level and that all odors are initially encoded as 'objects' in the piriform cortex. Moreover, we suggest that odor perception is wholly dependent on the integrity of this memory system and that its loss severely impairs normal perception. PMID- 12744841 TI - Ischemic tolerance and endogenous neuroprotection. AB - Practically any stimulus capable of causing injury to a tissue or organ can, when applied close to (but below) the threshold of damage, activate endogenous protective mechanisms--thus potentially lessening the impact of subsequent, more severe stimuli. A sub-threshold ischemic insult applied to the brain, for example, activates certain cellular pathways that can help to reduce damage caused by subsequent ischemic episodes--a phenomenon known as 'ischemic preconditioning' (IP) or 'ischemic tolerance' (IT). Although investigated for some time in model organisms, IP/IT has recently been shown in human brain. This opens a window into endogenous neuroprotection and, potentially, a window of opportunity to utilize these mechanisms in the clinic to treat patients with stroke and other CNS disorders. PMID- 12744843 TI - Genetic control of dendritic morphogenesis in Drosophila. AB - How the dendritic branching patterns of different neurons are specified is a fascinating question in developmental neurobiology. This question can now be addressed in detail in Drosophila, owing to technological advances that allow in vivo labeling of the dendrites of identifiable neurons. Recent genetic analyses in flies have uncovered several molecules, including transcription factors, cytoskeleton-associated proteins and membrane receptor-like molecules, that provide a glimpse into the complex regulatory network that controls dendritic morphogenesis. PMID- 12744842 TI - Talking back: dendritic neurotransmitter release. AB - Classical transmitters and neuropeptides can be released from the dendrites of many neuronal populations, to act as retrograde signals that modulate synaptic transmission, electrical activity and, in some cases, morphology of the cell of origin. For the hypothalamic neuroendocrine cells that release vasopressin and oxytocin, the stimuli, mechanisms and physiological functions of dendritic release have been revealed in detail that is not yet available for other neurons. The regulation of dendritic transmitter release is complex and at least partially independent from axon terminal release. Here, we provide an overview of recent findings on the mechanisms and physiological consequences of dendritic neuropeptide release and place this in the context of discoveries of dendritic neurotransmitter release in other brain regions. PMID- 12744844 TI - Sophisticated spinal contributions to motor control. AB - The neural circuitry of the spinal cord is capable of solving some of the most complex problems in motor control. Therefore, spinal mechanisms are much more sophisticated than many neuroscientists give them credit for. A key issue in motor control is how sensory inputs direct and inform motor output,--that is, the sensorimotor process. Other major issues involve the actual control of the motor apparatus. In general, there are at least three basic requirements for motor control: the transformations that map information from sensory to motor coordinates, the specification of individual muscle activations to achieve a kinematic goal, and the control of multiple degrees of freedom. Here, we make the case that the vertebrate spinal cord has the capacity to solve each of these problems to a degree that is relevant for normal behavior. PMID- 12744845 TI - Shedding light on brain mapping: advances in human optical imaging. AB - Several functional brain imaging techniques have been used to study human cortical organization. Optical imaging of intrinsic signals (OIS) offers perhaps the best combination of spatial coverage, resolution and speed for mapping the functional topography of human cortex. In this review, we discuss recent advances in optical imaging technology and methodology that have made human OIS easier to implement and more accessible, including improvements in detector characteristics and the development of sophisticated algorithms for reducing motion artifact. Moreover, we discuss how these advances have helped enhance our understanding of the functional organization of the human brain. We also review newly developed analyses for interpreting and validating optical signals, including refined signal analysis techniques and multimodality comparisons. Combined, these advances have enabled the study of not only primary sensory and motor cortices, but also higher cognitive processes such as language production and comprehension. Continued improvement and implementation of this technique promises to shed new light on the functional organization of human cortex. PMID- 12744846 TI - The use of hepatitis A vaccine in Italy--evidence-based recommendations from an expert panel. PMID- 12744848 TI - Hepatitis A: post-exposure prophylaxis. AB - We report on the findings of an exploratory review of evidence published in English from 1945 to identify the best post-exposure prophylaxis treatment and the longest acceptable interval after exposure for prophylaxis to be effective. We found no evidence that post-exposure administration of currently available immunoglobulins is effective in preventing hepatitis A infection and disease. The use of immunoglobulins for immunoprophylaxis should not be widely recommended until a systematic review of the evidence has been conducted. We recommend that active immunization to secondary contacts of exposed and vaccinated subjects be offered. PMID- 12744847 TI - Risk groups for hepatitis A virus infection. AB - We report the conduct and results of a systematic search for evidence of risk of infection with hepatitis A virus (HAV) among blood transfusion recipients, travellers, the military, healthcare workers, sewage workers, foodhandlers, day care assistants, institutionalised subjects, blood transfusion recipients, drug addicts, homosexuals, prisoners and other risk groups such a liver transplantees. We report our recommendations for the use of the HAV vaccine in these groups. PMID- 12744849 TI - HAV infection in chronic liver disease: a rationale for vaccination. AB - We report the conduct and results of a systematic search of the literature to assess whether exposure to HAV could induce a fatal deterioration of liver diseases. We assessed 30 studies and found evidence of progressive decrease of natural immunity against HAV in Italy. HAV vaccine seems as safe in chronic liver diseases than in general population. On the basis of the evidence we recommend that subjects with chronic liver disease should be vaccinated against HAV, especially if younger than 40 years. PMID- 12744850 TI - The effectiveness and safety of hepatitis A vaccine: a systematic review. AB - We report on the conduct of a systematic review to assess the efficacy and the safety of hepatitis A vaccines in adults and children. We identified, retrieved, and assessed all trials evaluating the effects of hepatitis A vaccines on prevention of cases of hepatitis A, death from hepatitis A, and assessing nature and frequency of adverse events. We included eight randomised trials, four containing efficacy outcomes, three containing only safety outcomes and a single study containing efficacy and adverse events outcomes. Combined inactivated vaccine effectiveness was 86% (95% CI: 63-95%). Combined attenuated vaccine effectiveness was 95% (95% CI: 81-99%). Inactivated vaccine effectiveness in the prevention of HAV secondary cases, compared to non-intervention was 82% (95% CI: 23-96%). Safety profile of vaccines was similar to that of their comparators. Despite poor design and reporting of trials, we found convincing evidence of the effectiveness and safety of inactivated HAV vaccines. PMID- 12744851 TI - The role of vaccine in controlling hepatitis A epidemics. AB - We report the conduct and results of a review of recent literature to describe various types of epidemics and outbreaks in Italy or countries with similar epidemiological profiles and to assess vaccine use in outbreak situations. We identified three scenarios most likely to occur in Italy: outbreaks occurring in small closed communities (nursery or a primary school), outbreaks in communities of limited dimensions (small towns or villages) and open community settings in which epidemics occur at regular intervals (person-to-person transmission). In closed communities we recommend vaccination of family members and school personnel living or working in close proximity to the index case as well as classmates. We also recommend vaccination when there is a proof of secondary transmission within the community. In small open communities we recommend vaccination of more susceptible age groups such as children and adolescents. For large open communities the only practicable alternative strategy is vaccination of close family contacts of acute cases couple with non-immunity boosting control measures. PMID- 12744852 TI - The use of hepatitis a vaccination in Italy: an economic evaluation. AB - We report on the conduct and results of an economic model of two different strategies of vaccination against hepatitis A in Italy. In strategy 1 (universal vaccination), all newborns are vaccinated simultaneously with MMR at 15 months and all 12 year olds are vaccinated with combined hepatitis A and B vaccine. In strategy 2 (contacts' vaccination) only subjects with a close social and spatial relationship with the sick are vaccinated. In strategy 1 costs per avoided case and net costs per avoided case values are always very high. Net costs become very low when newborns and 12 year olds are vaccinated, and become negative (indicating a potential gain) when 12 year olds are vaccinated. From an economic viewpoint, universal vaccination appears recommendable only in epidemic conditions. For strategy 2 our model shows that cost for avoidable case are very low and net costs for avoided case are always negative, showing the potential presence of an absolute benefit. We conclude that in Italy mass vaccination appears economically worthwhile only during epidemics but vaccination of contacts is an economically worthwhile routine measure. PMID- 12744853 TI - Simian immunodeficiency virus: possible effects of deworming and tuberculin extrapolated to HIV/AIDS. PMID- 12744854 TI - Comments on the paper "the effectiveness of serogroup C meningococcal vaccine estimated from routine surveillance data" [P Rivest, R Allard, Vaccine 20 (2002) 2533-2536]. PMID- 12744856 TI - Genetic immunization is augmented by murine polyomavirus VP1 pseudocapsids. AB - To improve immune responses induced by DNA immunization, murine polyomavirus major capsid protein (VP1) pseudocapsids were complexed with a DNA plasmid encoding the p37 (p24 and p17) nucleocapsid proteins of the human immunodeficiency virus type 1 (HIV-1). A 10-fold increase in antibody titer was noted in mice given DNA plasmid together with VP1 pseudocapsids in comparison to animals that received DNA plasmid alone. Cell mediated responses to HIV-1 p24 occurred, but were not significantly augmented by delivering the DNA as a VP1 complex. We have consequently for the first time shown a carrier/adjuvant effect of polyomavirus pseudocapsids that strongly increased the humoral immune response in DNA immunization. PMID- 12744858 TI - Development and clinical testing of multivalent vaccines based on a diphtheria tetanus-acellular pertussis vaccine: difficulties encountered and lessons learned. AB - The widespread use of whole-cell pertussis vaccines in the second half of the 20th century have reduced the incidence of the disease significantly. However, in some countries, concerns about the reactogenicity and potential neurological damage associated with whole-cell vaccines led to a decrease in vaccine acceptance and an increase in morbidity and mortality of pertussis in several countries. This prompted the development of less reactogenic acellular pertussis vaccines combined with diphtheria and tetanus toxoids, initially in Japan and later in other countries. In Europe, the improved diphtheria, tetanus and acellular pertussis (DTPa) vaccine was first introduced in March 1994. The pertussis component of this DTPa vaccine, developed by Glaxo SmithKline, consists of pertussis toxoid, filamentous haemagglutinin and pertactin. The vaccine is well tolerated, with a lower incidence of adverse reactions than after administration of whole-cell vaccines. The long-lasting efficacy and effectiveness of DTPa vaccines have been extensively documented and these are now the cornerstone of a large range of combined vaccines including DTPa-hepatitis B (HBV), DTPa-inactivated polio (IPV) and DTPa-HBV-IPV. A lyophilised Haemophilus influenzae type b (Hib) vaccine can be reconstituted with all of these liquid combinations. The introduction of well-tolerated and efficacious DTPa vaccines and their more polyvalent combinations has improved the acceptance and simplified the implementation of childhood immunisation. This paper is a review of the technical and scientific difficulties encountered and the lessons learned over the 10-year period that it took to develop and introduce six multivalent vaccines using the Glaxo SmithKline DTPa as a building block. PMID- 12744859 TI - Cellular immune responses to helper-free HSV-1 amplicon particles encoding HIV-1 gp120 are enhanced by DNA priming. AB - A single inoculation of herpes simplex virus type-1 (HSV-1) amplicon vectors encoding human immunodeficiency virus type-1 gp120 (HSV:gp120) results in robust, specific immune responses to gp120. To explore further the utility of this novel vaccine delivery system, we examined the kinetics of the cellular immune response by tetramer staining, following a single intramuscular administration of HSV:gp120 particles, and found that it peaks at 9-28 days post-immunization, before declining to a stable memory response. We also examined the utility of prime-boost regimens using packaged amplicon particles and naked amplicon plasmid DNA (DNA:gp120). These experiments showed that two sequential immunizations with HSV:gp120 resulted in a 5-10-fold increase in gp120-specific cellular immune responses and that plasmid DNA priming, followed by amplicon particle boosting, imparted the strongest acute and memory T cell responses, as determined by tetramer analysis. Collectively, these results demonstrate the utility of HSV amplicon vectors in prime-boost regimens for HIV vaccine development. PMID- 12744857 TI - CD40-targeted adenoviral gene transfer to dendritic cells through the use of a novel bispecific single-chain Fv antibody enhances cytotoxic T cell activation. AB - Adenoviral (Ad) transduction of dendritic cells (DC) is a promising vaccination strategy. However, clinical applicability of Ad vectors is hampered by the necessity to use high titers of infectious Ad particles for efficient DC transduction. Here, we report on the production of a bacterially expressed bispecific conjugate, consisting of a fusion of recombinant single-chain (sc) mAb Fv fragments, which bind and neutralize the Ad fiber knob (through the S11 mAb scFv) and retarget Ad to CD40 on the DC surface (through the G28-5 mAb scFv). We show that this bispecific scFv fusion protein significantly enhances transduction efficiency of monocyte-derived DC (MoDC), reduces the amount of virus needed for a given level of transduction, and increases the ability of MoDC to activate CTL in an antigen specific manner. This single-component conjugate may prove to be a valuable immunotherapeutic tool for the targeting of Ad to DC in vivo. PMID- 12744860 TI - Nature, evolution, and appraisal of adverse events and antibody response associated with the fifth consecutive dose of a five-component acellular pertussis-based combination vaccine. AB - We performed a randomized, controlled clinical trial to characterize the evolution of the adverse events associated with the fifth consecutive dose of an acellular pertussis vaccine, and to assess the level of discomfort associated with the injection and the attitude of parents concerning these events. A total of 505 children who had received either four doses of acellular pertussis vaccine or whole-cell pertussis vaccine were given a fifth dose of one of the two vaccines. Adverse events were monitored by parents and collected by telephone or home visit at 4, 8, 12, 24, 48 and 72 h, and 7 and 28 days after immunization. Rates of injection site redness >or=50mm were similar in recipients of five doses of acellular pertussis vaccine (32.8%) or five doses of whole-cell pertussis vaccine (43.3%). Injection site swelling, tenderness, and decreased arm movement were all more frequent in children who received five doses of whole-cell pertussis vaccine. Antibody levels before or after immunization did not predict those children who had increased injection site reactions. The children rated the injection site reactions as significantly more severe after five consecutive doses of whole-cell vaccine. Parent satisfaction was higher after the acellular vaccine. We conclude that a fifth consecutive dose of a whole-cell pertussis vaccine is associated with high rates of tender redness and swelling at the injection site, in contrast to a fifth consecutive dose of an acellular pertussis vaccine which is associated with high rates of non-painful redness. However, parents will still need to be aware of the high rates of injection site reactions expected after a fifth dose of acellular pertussis vaccine. PMID- 12744861 TI - Gene expression after vaccination of mice with formulations of diphtheria toxoid or tetanus toxoid and different adjuvants: identification of shared and vaccine specific genes in spleen lymphocytes. AB - We immunized mice with four different combinations of diphtheria toxoid or tetanus toxoid with aluminum phosphate or Freund's adjuvant and studied the resulting gene expression profiles in spleen lymphocytes. Genes, which are unique for each combination or shared in several combinations, were found activated, with functions in immune response but also in other cellular processes like apoptosis or signal transduction. Using bioinformatic tools we show, that some of the genes may serve as indicators for adverse reactions, while other genes may be new immune response markers. The results also suggest that adjuvant participates in the formation of an immunological memory. PMID- 12744862 TI - Mechanisms of enhanced antigen-specific T cell response following vaccination with a novel peptide-based cancer vaccine and systemic interleukin-2 (IL-2). AB - Systemic interleukin-2 (IL-2) therapy has been shown to enhance the clinical efficacy of peptide-based cancer vaccines. However, the mechanisms involved in this complex response remain poorly defined. IL-2 is known to be a potent T cell growth factor, but recent studies suggest that IL-2 is also involved in the regulation of T cell immune responses by increasing the susceptibility of proliferating T cells to apoptosis. Using an adoptive transfer model, we demonstrate that the administration of systemic IL-2 significantly enhances the primary and memory immune responses following peptide-based vaccination. In order to define the mechanisms of IL-2 therapy on the antigen-specific T cell response, the kinetics of T cell proliferation, apoptosis, and trafficking were explored. Systemic IL-2 therapy did not appear to alter the kinetics of T cell proliferation immediately following vaccination, but did prolong the proliferative response. Furthermore, IL-2 therapy did not significantly influence apoptosis of proliferating T cells. Such therapy did, however, potentiate L selectin (CD62L) downregulation on activated antigen-specific T cells, and altered their trafficking confirming their potential therapeutic value. Our findings support the use of systemic IL-2 following peptide-based vaccination, and suggest that IL-2 therapy enhances the primary and memory immune responses by prolonging the proliferative response and altering the trafficking of antigen specific T cells. PMID- 12744863 TI - Modifications of the catalytic and binding subunits of pertussis toxin by formaldehyde: effects on toxicity and immunogenicity. AB - A panel of pertussis toxin (PT) preparations with varying levels of residual toxicity was prepared by treatment of native PT with formaldehyde (0-1.00% (w/v)) with the purpose of investigating the effects of residual toxicity on immunogenicity. The catalytically inactive mutant PT (PT-9K/129G) was used for comparison. Results from in vitro ADP-ribosyl transferase and Chinese hamster ovary (CHO)-cell toxicity assays demonstrated a formaldehyde-dependent reduction in PT toxicity, and implied that both A and B domain functions of PT were modified. The in vivo histamine sensitisation and leukocyte proliferation tests suggested that the formaldehyde-treated native PT preparations were subject to reversion to toxicity. Reversion was confirmed by in vitro toxicity assays, which demonstrated recovery of A and B domain functions. The presence of high molecular weight aggregated and cross-linked species of PT in these preparations did not appear to be detrimental to the production of a neutralising antibody response. IgG responses to native and non-catalytic mutant PT suggested that low levels of residual activity in the native PT enhanced the antibody response, while higher levels of activity inhibited the response. Using the non-catalytic mutant PT showed that formaldehyde-induced changes were not detrimental to the magnitude of the PT-specific antibody response but did reduce the PT-specific neutralising activity. In conclusion, the residual toxicity of PT preparations following formaldehyde treatment may play an important role in the immune response to pertussis vaccine, potentially altering the quality, class and magnitude of the antibodies produced to PT. PMID- 12744864 TI - Immunogenicity of a Japanese encephalitis DNA vaccine candidate in cynomolgus monkeys. AB - A Japanese encephalitis (JE) vaccine candidate encoding JE virus premembrane (prM) and envelope (E) genes, designated pNJEME, was evaluated for safety and immunogenicity in non-human primate, cynomolgus monkeys. pNJEME was constructed using a vector (pNGVL4a) designed to address some of the safety concerns of DNA vaccine. In two different experiments, two immunizations with 300 microg of pNJEME by intramuscular (i.m.) injection, and 3 microg of pNJEME using a gene gun, and three immunizations by i.m. injection with 500 microg of pNJEME were performed. All the three protocols induced low to high levels of neutralizing antibody, indicating an ability of pNJEME to induce neutralizing antibody in monkeys with a wide individual variation in response to pNJEME. In one experiment designed to compare the DNA vaccine with a commercial inactivated JE vaccine, three immunizations by i.m. inoculation with 300 microg of pNJEME or by gene gun administration with 3 microg of pNJEME induced similar levels of neutralizing antibody to those induced by three immunizations with a human dose of the inactivated vaccine in most monkeys. After intranasal challenge with the Beijing P3 or JaTH160 strain of JE virus, pNJEME-immunized monkeys showed anamnestic neutralizing antibody responses, indicating that pNJEME induced memory B cells which were responsive to infection with JE virus. No systemic and local reactions were observed in any monkeys after i.m. or gene gun inoculations with plasmid DNAs. PMID- 12744865 TI - Oculo-respiratory syndrome following influenza vaccination: evidence for occurrence with more than one influenza vaccine. AB - We assessed the occurrence of oculo-respiratory syndrome (ORS) following two influenza vaccines: Fluviral (Shire Biologics) or Vaxigrip (Aventis Pasteur). ORS was identified amongst 5.3 and 4.6% of recipients, respectively (P=0.54). With both vaccines, the risk of ORS was much greater in individuals who had ORS the previous year (2000) than in those without such history. In multivariate analysis, the odds ratio for ORS for patients with a prior history of ORS varied between 9.4 and 9.6 (P<0.001) whereas that comparing Fluviral and Vaxigrip varied between 1.5 and 1.9 (P=0.02-0.05). ORS is an adverse event that is present with more than one vaccine and may be present with any influenza vaccines to a greater or lesser degree. PMID- 12744866 TI - The clinical spectrum of the oculo-respiratory syndrome after influenza vaccination. AB - Oculo-respiratory syndrome (ORS), a new influenza vaccine associated adverse event, was identified in 2000. The 2000 case definition (ORS-2000) required the presence of bilateral red eyes or respiratory symptoms or facial edema occurring between 2 and 24h following immunization and lasting 24 h), ORS persistors (duration >48 h).Overall, the distribution of symptoms was similar between ORS-2000 and other case categories. ORS-early and ORS-late had less ocular involvement, ORS-late and ORS-persistors had more cough and sore throat, ORS-early had more facial edema and ORS-late had less. In comparison to ORS-2000, ORS-early were younger whereas ORS-persistors and ORS-late were significantly older suggesting that clinical manifestations of ORS vary with age with a more rapid induction of symptoms in younger individuals and longer duration for older ones. PMID- 12744868 TI - Optimization of DNA vaccination immune responses in dairy cows: effect of injection site and the targeting efficacy of antigen-bCTLA-4 complex. AB - The objectives of this study were to evaluate the effects of immunization site and antigen presenting cell targeting on cattle immune responses to DNA immunization. Cows were vaccinated with the plasmid expression vector pCI alone, pCI encoding the bacterial antigen beta-galactosidase (pCI-beta-gal) or pCI encoding bCTLA 4 fused to beta-gal (pCI-bCTLA-hIgG-beta-gal). The plasmids were delivered by intramuscular, intradermal, intramammary gland, or intra supramammary lymph node needle-injection. Both vaccines induced significant humoral and cellular immune responses. pCI-beta-gal elicited a higher IgG response than immunization with pCI-bCTLA-hIgG-beta-gal. Cows injected intramuscularly and intramammary had higher IgG and IgG-1 humoral responses than cows immunized intradermaly or in the lymph nodes. The injection site did not significantly affect the magnitude of the IgG2 and IgM antibody responses, although a trend similar to the IgG results was observed. The lymphocyte proliferation index was higher with pCI-beta-gal but was not affected by the injection site. These results suggest that in bovine, the injection site can affect immune responses but they do not provide evidence that bCTLA-4-hIgG antigen targeting is effective in cattle. PMID- 12744867 TI - Roles of anti-hemagglutinin IgA and IgG antibodies in different sites of the respiratory tract of vaccinated mice in preventing lethal influenza pneumonia. AB - The roles of IgA and IgG antibodies (Abs) against hemagglutinin (HA) in the prevention of lethal influenza pneumonia in vaccinated mice were examined in terms of distribution and concentration of the Abs in the mucus or the serous fluid in different sites of the respiratory tract (RT), mucosa of the nose, trachea, bronchi and bronchioli and the alveolar epithelia of pulmonary acinus. First, the surface areas of the tracheal, bronchial and bronchiolar mucosa and alveolar epithelia were measured to be 20, 260 and 217, 433 mm(2), respectively, using serial tissue sections of the trachea and lungs. Then, the volumes of the tracheal mucus, the bronchial and bronchiolar mucus and the serous fluid of alveolar epithelia were estimated to be 0.2, 2.6 and 21.7 mm(3), respectively, by calculating each from the surface area and an assumed thickness of the mucus layer (0.01 mm) or that of the serous fluid (0.0001 mm). Next, anti-HA IgA and IgG Ab responses in the nasal wash, the trachea-lung wash and the trachea wash were measured in BALB/c mice immunized intranasally with an adjuvant-combined A/PR/8/34 (H1N1) virus vaccine and challenged with a lethal dose of the virus. Then the values of Ab responses were converted to the mucus and serous fluid Ab concentration based on two premises that the serum Abs diffuse at a constant rate to the surface of the tracheal, bronchial and bronchiolar mucosa, and that the active transepithelial transport of IgA Abs does not work in the alveolar epithelia. Results showed that 21.4 microg/ml IgA Abs and 3.6 microg/ml IgG Abs in the tracheal mucus (19.1 and 0.3% of the trachea-lung wash IgA and IgG Ab amounts, respectively), 5.9 microg/ml IgA Abs and 3.6 microg/ml IgG Abs in the bronchial and bronchiolar mucus (66.0 and 3.4% of the trachea-lung wash IgA and IgG Ab amounts, respectively) and about 0.1 microg/ml IgA Abs and 12.3 microg/ml IgG Abs in the serous fluid of alveolar epithelia (14.9 and 96.3% of the trachea lung wash IgA and IgG Ab amounts, respectively) were present in the vaccinated mice, at which concentrations influenza pneumonia was prevented. Thus, 96.3% of anti-HA IgG Abs in the trachea-lung wash work on the alveolar epithelia, whose surface area is about 800 times larger than that of tracheal, bronchial and bronchiolar mucosa and seem to play a more important role than the mucosal IgA Abs in the prevention of lethal influenza pneumonia. PMID- 12744869 TI - Antigen entrapped in the escheriosomes leads to the generation of CD4(+) helper and CD8(+) cytotoxic T cell response. AB - In previous study, we demonstrated the potential of Escherichia coli (E. coli) lipid liposomes (escheriosomes) to undergo membrane-membrane fusion with cytoplasmic membrane of the target cells including professional antigen presenting cells. Our present study demonstrates that antigen encapsulated in escheriosomes could be successfully delivered simultaneously to the cytosolic as well as endosomal processing pathways of antigen presenting cells, leading to the generation of both CD4(+) T-helper and CD8(+) cytotoxic T cell response. In contrast, encapsulation of same antigen in egg phosphatidyl-choline (egg PC) liposomes, just like antigen-incomplete Freund's adjuvant (IFA) complex, has inefficient access to the cytosolic pathway of MHC I-dependent antigen presentation and failed to generate antigen-specific CD8(+) cytotoxic T cell response. However, both egg PC liposomes as well as escheriosomes-encapsulated antigen elicited strong humoral immune response in immunized animals but antibody titre was significantly higher in the group of animals immunized with escheriosomes-encapsulated antigen. These results imply usage of liposome-based adjuvant as potential candidate vaccine capable of eliciting both cell-mediated as well as humoral immune responses. Furthermore, antigen entrapped in escheriosomes stimulates antigen-specific CD4(+) T cell proliferation and also enhances the level of IL-2, IFN-gamma and IL-4 in the immunized animals. PMID- 12744870 TI - Safety and immunogenicity of an oral, inactivated enterotoxigenic Escherichia coli plus cholera toxin B subunit vaccine in Bangladeshi children 18-36 months of age. AB - A phase II safety and immunogenicity study of an oral-formalin inactivated enterotoxigenic Escherichia coli (ETEC) vaccine containing six colonization factors (CFA/I, CS1, CS2, CS3, CS4, CS5) and 1mg of recombinant cholera toxin B subunit (the CF-BS-ETEC vaccine) was carried out in an urban slum of Dhaka city in Bangladesh. The study was carried out in a double blinded, placebo controlled design in 158 children, 18-36 months of age. Children were given two doses of the CF-BS-ETEC vaccine or the placebo which consisted of E. coli K12. The vaccine was well tolerated. The immune response was studied in 60 children (30 each in the placebo and vaccine group). Significant vaccine specific IgA antibody-secreting cell (ASC) responses were seen 7 days after ingestion of the first and second dose of the vaccine. The responses to CFA/I (Por=10 mIU/ml) was noted for 0, 25, 75, and 87.5% of subjects by increasing ISS dose group (P<0.05) for those who received ISS+HBsAg; 1 month after the second dose this increased to 62.5, 100, 100, and 100%, respectively. Geometric mean anti-HBsAg antibody levels by increasing ISS+HBsAg dose were 1.22, 5.78, 24.75, and 206.5 mIU/ml after the first dose and 65.37, 877.6, 1545, and 3045 mIU/ml after the second dose. We conclude that 1018 ISS+HBsAg was well tolerated and immunogenic in this phase I study in healthy adults and may offer the potential for enhancement of hepatitis B virus (HBV) immunization and protection after one or two doses or in individuals who fail to respond to the standard vaccine regimen. PMID- 12744880 TI - Analysis of PorA variable region 3 in meningococci: implications for vaccine policy? AB - Outer membrane protein (OMP) vaccines are being developed against Neisseria meningitidis serogroup B which may provide protection against common circulating serotypes and serosubtypes in some countries. However, limited data is available in Europe from genosubtyping meningococci. We therefore undertook a retrospective analysis of the three main variable regions, VR1, VR2 as well as VR3, of the porA gene from N. meningitidis isolated from different countries, mainly from Scotland and Sweden. Analysis of this gene showed that, amongst 226 strains studied, there were a total of 78 different strains. No new VR1 or VR2 alleles were found but five new VR3 alleles are described. Our data indicates the importance of analysing the VR3 region of PorA in addition to VR1 and VR2 and also highlights, in general terms, the need for genosubtyping meningococci. Such analyses have major implications for the design of new meningococcal vaccines. PMID- 12744881 TI - Protection in dogs against visceral leishmaniasis caused by Leishmania infantum is achieved by immunization with a heterologous prime-boost regime using DNA and vaccinia recombinant vectors expressing LACK. AB - A heterologous prime-boost vaccination regime with DNA and recombinant vaccinia virus (rVV) vectors expressing relevant antigens has been shown to enhance specific cellular immune responses and to elicit protection against a variety of pathogens in animal models. In this paper, we describe the effectiveness of the prime-boost strategy by immunizing dogs with a plasmid carrying the gene for the LACK antigen from Leishmania infantum (DNA-LACK) followed by a booster with a rVV containing the same gene (rVV-LACK). Thereafter, animals were challenged with L. infantum to induce visceral leishmaniasis (VL). In the vaccinated dogs as compared with the controls, the outcome of the infection after challenge with a high inoculum (10(8)) of L. infantum stationary promastigotes was assessed by tissue parasite load, specific anti-Leishmania antibody production, cytokine level and development of clinical signs of leishmaniasis. We observed a 60% protection against infection in dogs immunized by DNA-LACK prime/rVV/-LACK boost while two doses of DNA-LACK did not elicit protection against the disease. The interleukin 4 (IL-4), interferon gamma (IFNgamma) and IL-12 (p40 subunit) cytokine mRNA expression profiles in PBMC as well as lymphocyte proliferative response and the IgG2/IgG1 ratios specific for LACK suggest that in vaccinated animals there is triggering of cellular immune responses. This type of DNA/rVV prime/boost immunization approach may have utility against visceral leishmaniasis in dogs. PMID- 12744882 TI - OM-174, a new adjuvant with a potential for human use, induces a protective response when administered with the synthetic C-terminal fragment 242-310 from the circumsporozoite protein of Plasmodium berghei. AB - The goal of this project was the evaluation of a novel immunomodulatory adjuvant for human use, OM-174, which is a soluble adjuvant derived from Escherichia coli lipid A. For this study, we used a synthetic peptide, known for its safety and reproducibility and the murine model of BALB/c mice. The long peptide (PbCS 242 310) used corresponds to the C-terminal region of the circumsporozoite protein (CSP) that is the major protein on the surface of Plasmodium sporozoites. Subcutaneous injections of PbCS 242-310 in combination with soluble adjuvant OM 174 induced long lasting peptide-specific antibody titres comparable to those obtained by immunization with incomplete Freund's adjuvant (IFA). The ex vivo evaluation of the CD8(+) T cell response by IFN-gamma ELISPOT assay revealed that the injection of polypeptide with OM-174 adjuvant induced, compared to IFA, a similar and an eight-fold increased frequency of peptide-specific lymphocytes in the draining lymph-nodes and in the spleen, respectively. The CD8(+) T-cells are specific for the sequence PbCS 245-253, a well-known H-2K(d)-restricted CTL epitope, and are cytotoxic as shown in a chromium release assay. Immunization of BALB/c mice with this polypeptide in combination with adjuvant OM-174 conferred a protection after challenge with live Plasmodium berghei sporozoites.The strong antibody and CTL responses observed to a synthetic peptide in mice, the safety profile of the adjuvant and its extensive physico-chemical characterization suggest that OM-174 has a potential use in vaccine formulations for humans. PMID- 12744883 TI - CpG-motifs enhance initial and sustained primary tetanus-specific antibody secreting cell responses in spleen and bone marrow, but are more effective in adult than in neonatal mice. AB - Unmethylated CpG oligonucleotides (CpG-ODN) increase adult and neonatal primary antibody responses to T-dependent antigens, at yet unidentified stages of antigen specific B cell differentiation. In adult mice, a single dose of CpG-ODN adjuvanted tetanus toxoid (TT) vaccine markedly enhanced and prolonged splenic TT specific antibody-secreting-cell (ASC) responses and significantly increased the size of the bone marrow (BM) ASC pool. Surprisingly, this was not associated with changes of germinal center (GC) numbers, size, apoptosis or function. In 1-week old mice, CpG-ODN also enhanced TT-specific splenic ASC responses, but failed to correct limitations of the GC reaction and of the development of the BM ASC pool. PMID- 12744884 TI - A single amino acid substitution in a recombinant G protein vaccine drastically curtails protective immunity against respiratory syncytial virus (RSV). AB - Recent studies have indicated a dominant T cell epitope located approximately between amino acids 184 and 203 on the respiratory syncytial virus (RSV) G protein. Using an Escherichia coli-grown plasmid vector encoding a fragment of thioredoxin (Trx) fused to a central region (amino acids 128-229) of the RSV G protein, we employed site-directed mutagenesis to investigate the importance of selected amino acids on vaccine efficacy. By changing two amino acids Arg 188 and Lys 192 to alanine, the ability of the Trx-G 128-229 fusion protein to protect mice against RSV challenge was virtually abolished. Mice immunized with the double mutant protein showed low levels of neutralizing antibodies and no pulmonary eosinophilic infiltrate, in contrast to that observed in mice immunized with wild type protein prior to RSV challenge. While less effective than the double mutant, mutation of either Arg 188 or Lys 192 to Ala drastically impaired the ability of immunized Trx-G 128-229 to induce neutralizing antibodies and to elicit pulmonary eosinophilia associated with RSV challenge. Despite low levels of virus-neutralizing antibodies, G protein-specific antibodies were detected by Western blotting in the sera from mice immunized with either of the single mutants (Arg 188 or Lys 192) but not the double mutant. Finally, immunization of mice with truncated forms of the Trx-G protein, showed partial protection against RSV challenge with Trx-G 128-188 but not with Trx-G 189-229. Taken together, the results indicate an important role for Arg 188 and Lys 192 in the induction of protective immunity and priming for eosinophilia against RSV. Furthermore, while the dominant protective linear epitope on the RSV G protein requires an intact sequence around Arg 188, there are additional, but less potent, protective epitopes upstream of Arg 188. PMID- 12744885 TI - Enhanced tumour growth after DNA vaccination against human papilloma virus E7 oncoprotein: evidence for tumour-induced immune deviation. AB - We have examined the induction of anti-tumour immunity in a murine model using a gene vaccine approach to deliver a well defined tumour antigen. The vaccines expressed the human papilloma virus type 16 (HPV 16) E7 oncoprotein, and protection was measured against HPV 16-expressing C3R tumour cell line in vivo. In control mice injected with saline, C3R cells initially formed tumours but then regressed completely. As expected, animals injected with a peptide that represents the D(b)-presented CTL epitope from E7 (RAHYNIVTF) were completely protected from tumour growth. Contrary to expectation, however, we consistently saw enhanced tumour growth, delayed regression, or tumour outgrowth in mice vaccinated with two different E7-expressing DNA vaccines. We found no evidence for loss of D(b) or K(b) class I MHC molecules from C3R cells recovered from outgrown tumours, and fluorescent MHC/peptide tetramer staining revealed E7 gene vaccination did not delete RAHYNIVTF-specific CD8(+) T cells. However, we did observe an effect on cytokine production. Splenocytes from E7 gene vaccinated animals responded to re-stimulation in vitro with C3R cells by producing IL-4 but background levels of IFN-gamma. We also observed that cytokine production and E7 peptide-specific CTL were only detectable in vaccinated animals after C3R challenge, but not after DNA priming alone. We conclude that 'prime-boosting' is necessary to observe tumour-specific T cell responses with the gene vaccine approach, but that boosting with tumour cells causes skewing of the primed cells in a T2 direction that is incompatible with protective anti-tumour immunity. PMID- 12744886 TI - The domain III fragment of Japanese encephalitis virus envelope protein: mouse immunogenicity and liposome adjuvanticity. AB - The E protein of Japanese encephalitis virus (JEV) is the major antigen used to elicit neutralizing antibody response and protective immunity in hosts. In this study, the domain III protein of the attenuated strain CH2195LA was cloned to the pET32a expression vector and expressed as a thioredoxin (Trx) fusion protein in Escherichia coli. The recombinant protein was unique in forming a large fraction of the soluble recombinant protein in E. coli. The purified domain III fusion protein (TrxD3) was emulsified in Freund's adjuvant (FA) as well as in different charged liposomes for immunization in mice. Immunization of TrxD3 fusion protein emulsified in Freund's adjuvant and only the cationic liposome resulted in eliciting neutralizing antibodies and protective immunity in ICR mice. The cationic liposome can serve not only as a safer but also an effective adjuvant for the TrxD3 protein immunization. These studies can provide useful information for further developing the domain III recombinant protein vaccine against JEV. PMID- 12744887 TI - Antibody response to vaccination and psychosocial stress in humans: relationships and mechanisms. AB - The purpose of this review is to determine the effects of psychosocial stress on antibody response to vaccination in humans, consider possible mechanisms, and identify agenda for future research. Studies of the association between stress and vaccination response in humans were reviewed. There is evidence of a negative association between stress and antibody response to vaccination, which is most apparent with thymus-dependent vaccines and when measured at extended times after vaccination. Preliminary findings implicate the hypothalamic-pituitary-adrenal axis and sympathetic nervous system as potential mechanisms, although a role for unhealthy behaviours cannot be discounted at this stage. Results to date are sufficiently indicative to direct future research to untangling their theoretical ramifications, as well as realising their clinical implications. PMID- 12744888 TI - Three types of immunotherapics against pythiosis insidiosi developed and evaluated. AB - Pythiosis is a granulomatous disease of horses, cattle, dogs, cats and humans identified in tropical and subtropical areas and caused by Pythium insidiosum, a zoosporic fungus. Experimental models of pythiosis in naturally infected species have not yet been reported but, rabbits may be inoculated with zoospores as an experimental model for studying the disease. The present study evaluates the efficacy of three different of immunotherapics in the rabbit model. Approximately 17500 zoospores of oomycete P. insidiosum (CBS 101555 strain) were inoculated in each animal to generate the disease. Immunotherapics were produced from vortexed or sonicated cultures of the same strain. Four groups of five animals were employed: group 1, placebo; group 2, sonicated immunotherapic; group 3, mixed immunotherapic; and group 4, vortexed immunotherapic. All rabbits were inoculated with viable zoospores one month before administration of the immunotherapics. Eight doses of immunotherapic or placebo were used in each animal with a 14 day interval between injections. Rabbits receiving the vortexed immunotherapic were most effectively protected (P<0.05), showing a decrease in the area of coastal nodules due to Pythiosis insidiosum by 71.8% after 26 weeks of evaluation. Moreover, two animals in this group showed complete remission of the infection at the end of the 26 weeks. In contrast to these findings, rabbits given the sonicated immunotherapic did not show any protection and had an increase of 211.8% in the size of lesions. This failure of sonicated immunotherapic may reflect denaturation of protective antigens due to the sonication method. PMID- 12744889 TI - The incidence of shingles and its implications for vaccination policy. AB - A vaccine is now available to prevent varicella-zoster infection, but its place in routine preventive care is not yet determined. The age specific incidence of shingles was examined separately by gender and age groups (15-24, 25-44, 45-64, 65-74 and 75 years and more) over the years 1994-2001. These incidence data were applied to national available data for the UK on current life expectancy to calculate the risk of shingles infections at varying ages. The potential benefit of an effective vaccine was estimated using three models of vaccine efficacy applied separately to males and females at ages 50, 60 and 65 years and assuming vaccination at a single age. Similar calculations were made using a two dose strategy at age 45 and 65 years and at age 50 and 70 years. The cost per case saved was estimated from a vaccination cost of pound 40 per dose. The probability of having had an attack of shingles before age 45 years is 8.6% for males and 10.5% for females, The risk of acquiring shingles over an expected lifetime (assuming no preventive vaccination) for males aged 45 years is 22% and for females 32%. Whichever vaccine efficacy model was chosen, a single vaccination policy at age 65 years was the most favourable option in both males and females. A two age vaccination policy was estimated to increase the cost per case saved by 30% over a single age policy but administration at age 50 and 70 years substantially increased the number of cases saved as compared with a single age policy and was potentially better than vaccination at 45 and 65 years. PMID- 12744890 TI - Murine antibody response to intranasally administered enterotoxigenic Escherichia coli colonization factor CS6. AB - Enterotoxigenic Escherichia coli (ETEC) is the most common cause of bacterial diarrhea worldwide and is an important cause of infant morbidity and mortality in developing nations. ETEC colonization factors (CF) are virulence determinants that appear to be protective antigens in humans and are the major target of vaccine efforts. One of the most prevalent CF, CS6, is expressed by about 30% of ETEC worldwide. This study was designed to compare the immunogenicity between encapsulated CS6 (CS6-PLG) and unencapsulated CS6. Recombinant CS6 was purified and encapsulated in biodegradable poly(DL-lactide-co-glycolide) (PLG) microspheres using current Good Manufacturing Practices (cGMP). CS6-PLG and CS6 were administered intranasally (IN) to BALB/c mice in three vaccinations 4 weeks apart. Enzyme linked immunosorbent assay (ELISA) was used to measure the anti-CS6 response in serum and mucosal secretions following each of the three inoculations. Mice vaccinated with two or three doses of CS6-PLG demonstrated a significantly greater rise in serum anti-CS6 IgG and mucosal IgA titer values than those immunized with two or three doses of CS6 alone. Three doses of CS6-PLG led to anti-CS6 serum IgG and mucosal IgA titer values 14-fold and 4.4-fold greater, respectively, than three doses of CS6 (P<0.02). IN administered CS6 to mice is safe and highly immunogenic either alone or when encapsulated in microspheres. PLG microsphere encapsulation of CS6 significantly augments the antibody response to that antigen when administered to a mucosal surface. PMID- 12744891 TI - Safety and protective efficacy of porcine reproductive and respiratory syndrome recombinant virus vaccines in young pigs. AB - Three porcine reproductive and respiratory syndrome virus (PRRSV) recombinants, generated by mutagenesis of an infectious cDNA clone of the Lelystad virus (LV) isolate, were tested for their safety and protective efficacy as potential PRRSV vaccines in pigs. Recombinant vABV688 contains two amino acid substitutions in the minor structural protein GP(2) resulting in improved growth on cell line CL2621; in recombinant vABV707 the region encoding the ectodomain of the major unglycosylated membrane protein M has been replaced by that of the murine lactate dehydrogenase-elevating arterivirus; recombinant vABV746 lacks the six C-terminal amino acids of the nucleocapsid protein N. First, we determined the safety of these recombinant viruses by monitoring the stability of the introduced mutations in 8-week-old pigs. We showed that the introduced genomic mutations were maintained throughout the viraemic period. Second, the protective efficacy of immunization with the recombinant viruses against challenge with a homologous and a heterologous PRRSV strain was determined in two pigs and compared with the efficacy of vABV437, a virus derived from the parental LV cDNA. The viraemia in pigs immunized with the recombinant viruses was reduced compared to pigs immunized with vABV437. In addition, the length of viraemia was reduced in the sentinel pigs that were introduced into the groups immunized with vABV746, vABV688, and vABV707, however, all of the sentinel pigs became infected. Pigs immunized with vABV707 and vABV437 were protected against challenge with homologous virus LV-Ter Huurne and transmission of the latter virus. None of the immunized pigs were protected against heterologous challenge with the virulent US isolate SDSU#73, but the vABV707- and vABV746-immunized pigs were protected against transmission of this virus from challenged pigs. In conclusion, the obtained viral recombinants are interesting candidates to be further explored for their use as vaccines against PRRSV. PMID- 12744892 TI - The cost-burden of paediatric pneumococcal disease in the UK and the potential cost-effectiveness of prevention using 7-valent pneumococcal conjugate vaccine. AB - We modelled the epidemiology and cost of pneumococcal disease in children in the UK and the cost-effectiveness of immunisation with 7-valent pneumococcal conjugate vaccine (PCV). We estimated the incidence of pneumococcal meningitis, pneumococcal septicaemia, all-cause pneumonia and all-cause otitis media (OM). We further estimated the impact of vaccination with associated costs and outcomes. Vaccine cost was pound 39.25 per dose with a pound 10 administration cost; vaccination schedule and efficacy were taken from a recent trial. We estimated that in each UK annual birth cohort there are 881,146 episodes of these infections and 149 deaths associated with pneumococcal meningitis, pneumococcal septicaemia or all-cause pneumonia and that PCV would prevent 54,384 episodes and 29 deaths. NHS cost per life year gained was estimated at pound 31,512, close to the limit at which PCV would be considered cost-effective. PMID- 12744893 TI - Effect of infant immunisation with meningococcus serogroup C-CRM(197) conjugate vaccine on diphtheria immunity and reactogenicity in pre-school aged children. AB - The majority of Men C conjugate vaccines given in the UK use CRM(197), a mutant diphtheria toxoid, as their protein carrier. We studied the effects of prior immunisation with Men C-CRM(197) conjugate vaccine on immunity to diphtheria in 193 children before and after a booster dose of Men C at 4 years. Baseline diphtheria antibodies were higher in children given four previous doses of Men C (P<0.0001) and tended to be higher following boosting in those who had received three or four doses. This enhanced immunity was not associated with increased reactogenicity. PMID- 12744895 TI - IgG1/IgG2 antibody dichotomy in sera of vaccinated or naturally infected dogs with visceral leishmaniosis. AB - Canine antibody IgG, IgG1 and IgG2 anti-FML responses were investigated in dogs vaccinated with the fucose-mannose ligand (FML)-vaccine of Leishmania donovani and in dogs with naturally acquired visceral leishmaniosis. While similar levels of total IgG antibodies were seen in the seropositive naturally infected dogs and in vaccinees, significant differences between the groups were found regarding their IgG1/IgG2 anti-FML antibody composition (P<0.005). Higher IgG1 absorbencies were seen in infected dogs, while the IgG2 subtype was predominant in pre-immune sera, and in vaccinated animals, both after the first and the third dose (P<0.005). The average ratio between IgG1/IgG2 was then 1.124 for infected animals and 0.733 for FML-vaccinees. Also, a significant increase in IgG2 antibodies was observed from the first to the third vaccine injection (P<0.005). In the infected dogs, a high correlation between their IgG absorbance (Abs) values and the number of symptoms (P=0.017) was disclosed. Thus, the analysis of IgG subclasses disclosed a dichotomous response to visceral leishmaniosis: IgG1 associated to natural infection and IgG2 associated to a humoral response subsequent to the FML-vaccine treatment. An IgG1/IgG2>or=1 would characterize the sera of visceral leishmaniasis infected animals evoluting towards the overt disease while ratios /= 25 kg/m(2); obesity as BMI >/= 30. Associations between sociodemographic characteristics, exercise frequency, health indicators, and weight-related measures were evaluated to identify independent predictors of BMI and exercise frequency. RESULTS: Prevalence of overweight and obesity among lesbians varied by racial/ethnic background. Higher BMI was associated with older age, poorer health status, lower educational attainment, relationship cohabitation, and lower exercise frequency. Higher BMI, perceptions of being overweight, and reporting a limiting health condition were identified as independent predictors of infrequent exercise. Women were generally quite accurate in self-perceptions of weight status. CONCLUSIONS: Correlates of overweight and obesity among lesbians and bisexual women are generally comparable to those observed in studies of heterosexual women. Evidence that lesbians' higher BMI is associated with higher levels of fitness is not supported. PMID- 12744912 TI - Association of blood pressure with intake of soy products and other food groups in Japanese men and women. AB - BACKGROUND: Soy diet has been suggested to have antihypertensive effect in animal studies. The present study examined the cross-sectional relationship between blood pressure and intake of soy products and other food groups in Japanese men and women. METHODS: Blood pressure was measured in Japanese 294 men and 330 women (246 premenopausal and 84 peri- and postmenopausal women) who participated in a health check-up program provided by a general hospital. Intake of various food groups and nutrients was estimated from a validated semiquantitative food frequency questionnaire. RESULTS: In men, soy product intake was inversely significantly correlated with diastolic blood pressure (r = -0.12, P = 0.04) after controlling for age, total energy, smoking status, body mass index, and intake of alcohol, salt and seaweeds. The correlation of soy product intake with systolic blood pressure was of borderline significance (r = -0.10, P = 0.09). Systolic blood pressure was inversely correlated with intake of vegetables (r = 0.12, P = 0.04) and dairy products (r = -0.12, P = 0.05). There were no significant correlations between soy product intake and diastolic blood pressure in women. CONCLUSIONS: These results indicate a mild effect of soy intake on blood pressure reduction in men. PMID- 12744913 TI - Association between physical activity and mental disorders among adults in the United States. AB - BACKGROUND: The objective of this study was to determine the association between regular physical activity and mental disorders among adults in the United States. METHODS: Multiple logistic regression analyses were used to compare the prevalence of mental disorders among those who did and did not report regular physical activity using data from the National Comorbidity Survey (n = 8098), a nationally representative sample of adults ages 15-54 in the United States. CONCLUSIONS: Slightly over one-half of adults reported regular physical activity (60.3%). Regular physical activity was associated with a significantly decreased prevalence of current major depression and anxiety disorders, but was not significantly associated with other affective, substance use, or psychotic disorders. The association between regular physical activity and lower prevalence of current major depression (OR = 0.75 (0.6,0.94)), panic attacks (OR = 0.73 (0.56, 0.96)), social phobia (OR = 0.65 (0.53, 0.8)), specific phobia (OR = 0.78 (0.63, 0.97)), and agoraphobia (OR = 0.64 (0.43, 0.94)) persisted after adjusting for differences in sociodemographic characteristics, self-reported physical disorders, and comorbid mental disorders. Self-reported frequency of physical activity also showed a dose-response relation with current mental disorders. DISCUSSION: These data document a negative association between regular physical activity and depressive and anxiety disorders among adults in the U.S. population. Future research that investigates the mechanism of this association using longitudinal data to examine the link between physical activity and incident and recurrent mental disorders across the lifespan is needed. PMID- 12744914 TI - Impact of lifestyle on perioperative smoking cessation and postoperative complication rate. AB - OBJECTIVE: The aim was to examine to what extent lifestyle, education, social support, and comorbidity predict the ability of perioperative smoking cessation, and are associated with the development of important postoperative complications. DESIGN: The design was a randomized clinical trial. SETTING: University hospitals in Copenhagen, Denmark, were the settings. PARTICIPANTS AND METHODS: One hundred twenty patients scheduled for primary elective hip or knee arthroplasty were randomized to either smoking intervention or standard care. Tobacco and alcohol consumption, exercise and eating habits, level of education, matrimonial status, and the presence of social support were registered. The data gathered concerned smoking cessation/reduction and severe postoperative morbidity. RESULTS: Men and patients with a good social network were more likely to successfully quit smoking. Smoking intervention successfully reduced the incidence of postoperative complications, as did weekly exercise exceeding 4 h, and having a high education level. CONCLUSIONS: This study emphasizes that smoking intervention programs in health care settings are highly effective in reducing postoperative risks in hip and knee arthroplasty. PMID- 12744915 TI - Distribution of daily smokers by stage of change: Current Population Survey results. AB - BACKGROUND: Population-based national estimates of stage of change among daily smokers are unknown. This study described the proportion of U.S. daily smokers, 18 and older, by stage of change. Selected sociodemographic characteristics were delineated. METHODS: Cross-sectional data were collected via telephone or face-to face interview in daily smokers who responded to the Current Population Survey in 1992-1993 (n = 39,706), 1995-1996 (n = 34,865), or 1998-1999 (n = 30,153). Main outcomes included stage of change: (1) Precontemplation-not interested in quitting smoking in next 6 months; (2) Contemplation-interested in quitting smoking in next 6 months but not next 30 days; (3) Preparation-interested in quitting smoking in next 30 days and stopped at least 1 day during past year. RESULTS: During 1992-1993, 59.1% of respondents were precontemplators, 33.2% contemplators, and 7.7% in preparation stage. This distribution was similar in subsequent surveys (1995-1996; 1998-1999). Gender differences were not apparent. Whites were more likely to be precontemplators. As education and income increased, the percentage in precontemplation decreased. Rural residents were more likely in precontemplation and less frequently in preparation. CONCLUSIONS: Among daily smokers, little movement in stage of change was apparent in the United States during the 1990s. Tobacco control efforts must receive high priority to address these static patterns. PMID- 12744917 TI - Internet cigarette purchasing among ninth-grade students in Western New York. PMID- 12744916 TI - Socioeconomic disparities in cancer-risk behaviors in adolescence: baseline results from the Health and Behaviour in Teenagers Study (HABITS). AB - BACKGROUND: This study explores the association between socioeconomic deprivation and five factors associated with long-term risk of cancer, in adolescents. METHODS: BMI, fat intake, fruit and vegetable intake, smoking, and exercise were assessed in 4320 students ages 11 to 12, from 36 schools, in the first year of a 5-year longitudinal study of the development of health behaviors (HABITS study). Neighborhood socioeconomic deprivation for each student's area of residence was matched to their postcode (zip code). We used multiple logistic regression analyses to investigate the relationship between risky behaviors and socioeconomic circumstances. RESULTS: Univariate analyses showed boys and girls from more deprived neighborhoods were more likely to have tried smoking, to eat a high fat diet, and to be overweight. Girls living in more deprived areas were also less likely to eat five servings of fruit and vegetables or to exercise at the weekend. Most differences persisted after controlling for ethnicity. A clear deprivation gradient emerged for each risk factor, indicating the linear nature of the relationship. CONCLUSIONS: This study demonstrates the influence of deprivation on engaging in cancer-risk health behaviors. These patterns may set young people from more socioeconomically deprived social environments on a trajectory leading to increased cancer mortality in adult life. PMID- 12744918 TI - Association between CBE, FOBT, and Pap smear adherence and mammography adherence among older low-income women. AB - BACKGROUND: Adherence to regular and timely mammography screening, especially in older low-income women, continues to fall below objectives. The primary aim of this study was to examine whether engaging in other cancer screenings was associated with mammography adherence for older women. METHODS: Women, ages 52 and over, without a self-reported history of breast cancer (N = 862) were selected from a larger sample of women residing in Washington, DC, census tracts with >/=30% of households below 200% of the federal poverty threshold. A computer assisted telephone survey was used to collect data on health care system factors, demographics, cultural beliefs, clinical breast exam (CBE), Pap smear, fecal occult blood testing (FOBT), and mammography. Adherence was defined as receipt of the last two screening tests within recommended intervals for age. RESULTS: After controlling for other variables, adherence to CBE (OR = 4.15; 95% CI, 2.55-6.73) and Pap smear (OR = 1.82; 95% CI, 1.07-3.12) were highly predictive of mammography adherence. Adherence to FOBT (OR = 1.66; 95% CI, 0.97-2.84) was marginally predictive. CONCLUSIONS: Results of this study indicate that nonadherence to other cancer screenings can help identify women in need of additional interventions to improve mammography adherence. PMID- 12744919 TI - Human placental trophoblast cells express alpha-tocopherol transfer protein. AB - Alpha-tocopherol transfer protein (alpha-TTP), a 30 kDa cytosolic protein first described to be present in the liver and important for alpha-tocopherol trafficking, plays a major role in maintaining alpha-tocopherol levels in plasma, while alpha-tocopherol is known as the major lipid-soluble antioxidant. Expression of alpha-TTP has not only been described in animal model liver, but also in diverse other tissues such as rat brain or pregnant mouse uterus, the latter finding stressing the importance of alpha-TTP for embryogenesis and foetal development. In this study, we report the identification of alpha-TTP in human liver by anti-human alpha-TTP monoclonal antibodies made in rat and the cellular localization of alpha-TTP in term human placenta. By immunohistochemistry, intense staining of alpha-TTP was seen in syncytiotrophoblast as well as in villous and invading extravillous cytotrophoblast, while basal decidual cells showed slighter, but present staining of alpha-TTP. Foetal vessel endothelium remained unstained. It is therefore suggested that alpha-TTP may play a major role in supplying alpha-tocopherol to the foetus prior to delivery and is likely involved in maintaining adequate alpha-tocopherol levels in the foetus. PMID- 12744920 TI - ATP dependent Ca2+ transport across basal membrane of human syncytiotrophoblast in pregnancies complicated by intrauterine growth restriction or diabetes. AB - Neonates born after pregnancies complicated by diabetes or intrauterine growth restriction (IUGR) have increased incidence of hypocalcaemia. Furthermore, IUGR is associated with reduced bone mineralization in infancy and osteoporosis in adult life. We tested the hypothesis that placental calcium transport is altered in these pregnancy complications. Transport of calcium into syncytiotrophoblast basal plasma membrane (BM) vesicles was studied by rapid filtration and protein expression of Ca(2+) ATPase by Western blot. In IUGR Ca(2+) ATPase activity was increased by 48 per cent (n=13; P< 0.05) whereas protein expression was 15 per cent lower (n=13; P< 0.05) than in controls (n=16). Basal membrane ATP dependent calcium transport was unaltered in gestational diabetes (GDM) but increased by 54 per cent in insulin dependent diabetes (IDDM) compared to controls (P< 0.05; n =14). Diabetes did not affect Ca(2+) ATPase expression in BM. We have previously shown that the mid-molecular fragment of parathyroid hormone related peptide (PTHrP midmolecule) stimulates BM Ca(2+) ATPase in vitro. PTHrP midmolecule concentrations in umbilical cord plasma were measured using radioimmunoassay. The concentrations in umbilical cord plasma were increased in IUGR, but unaltered in diabetes. In conclusion, placental calcium pump is activated in IUGR and IDDM, which may be secondary to increased foetal calcium demand. We speculate that PTHrP midmolecule may be one mechanism for activating BM Ca(2+) ATPase in IUGR. PMID- 12744921 TI - Localization of alkaline phosphatase and Ca2+-ATPase in the cat placenta. AB - We localized alkaline phosphatase and plasma membrane calcium-ATPase (PMCA) in the cat placental syncytiotrophoblast to address their polarized distribution and their potential as markers for specific plasma membrane purification. We used enzyme- (alkaline phosphatase) and immuno- (PMCA) histochemistry and, for alkaline phosphatase, compared data to observations on the human placenta. Alkaline phosphatase activity in the cat was localized to the decidual cell membranes, to within the associated interstitial space and on the subjacent apical (maternal facing) plasma membrane of the syncytiotrophoblast. Occasional maternal capillaries were positive on their basal surface and there was focal staining within the syncytiotrophoblast. This widespread distribution is less specific than in the human placenta where alkaline phosphatase was restricted to the apical and basal plasma syncytiotrophoblast membranes, with much greater density on the apical membrane. Expression of PMCA in the cat was restricted to the basal membrane of the syncytiotrophoblast only. This specific localization of PMCA is identical to the human placenta and all other species in which its placental localization has been studied. We conclude that the plasma membranes of the cat syncytiotrophoblast show a broadly similar functional polarization to the human and that PMCA would prove a useful marker in isolation of the cat syncytiotrophoblast basal plasma membrane. PMID- 12744922 TI - Excretion of foetal bilirubin by the rat placenta-maternal liver tandem. AB - Using plasma membrane vesicles from human trophoblast, carrier-mediated transport of unconjugated bilirubin (UCBR) has been reported. In the present work, using the in situ perfused rat placenta-maternal liver tandem, the relevance of this pathway in vivo was investigated. After single-pass perfusion of rat placenta through the umbilical artery with 0.25 micromol [(3)H]-UCBR, approximately 15 per cent of it was taken up by the placenta, detected in maternal serum (>96 per cent was unconjugated) and subsequently secreted into maternal bile (approximately 15 per cent of administered dose; >88 per cent was glucuronidated bilirubin). Co administration through the umbilical artery of 0.25 micromol [(3)H]-UCBR and 2.5 micromol unlabelled UCBR, bromosulfophthalein, cholic acid or biliverdin IXalpha, reduced [(3)H]-UCBR placenta uptake, and the amount of radioactivity found in the maternal serum and bile. Co-administration into maternal jugular vein of 0.1 micromol [(3)H]-UCBR-a dose 3-fold higher than that reaching the maternal compartment in placenta perfusion experiments-and 1.0 micromol bromosulfophthalein, cholic acid or biliverdin IXalpha, resulted in no marked inhibition of the amount of radioactivity bile output. When antipyrine and [(3)H] UCBR were continuously co-infused to the mother, similar antipyrine concentrations in maternal and foetal serum were reached in approximately 15 min, while progressive increase in [(3)H]-bilirubin concentrations in maternal serum above 70 microM was accompanied by a very low transfer of this compound into foetal compartment where [(3)H]-bilirubin concentrations were always <10 microM. These results suggest that the transfer of UCBR across the rat placenta occurs, without biotransformation, via a foetal-to-maternal mainly unidirectional pathway that can be cis-inhibited by UCBR and other cholephilic organic anions. PMID- 12744923 TI - Identification of the CB1 cannabinoid receptor and fatty acid amide hydrolase (FAAH) in the human placenta. AB - Synthetic cannabinoids, the psychoactive components of the Cannabis sativa (marijuana) and their endogenous counterparts, act through two G protein-coupled receptors, CB1 and CB2. The endocannabinoids are metabolized by fatty acid amide hydrolase (FAAH). Previous research has described the impact of cannabis consumption on pregnancy, potential roles of endocannabinoids and abnormalities of FAAH expression in recurrent miscarriage and pregnancy. However, the cellular localization of the CB1 cannabinoid receptor and FAAH in the human placenta has not been determined. We have examined CB1 receptor and FAAH expression in human term placenta by immunohistochemistry. CB1 receptor was found to be present in all layers of the membrane, with particularly strong expression in the amniotic epithelium and reticular cells and cells of the maternal decidua layer. Moderate expression was observed in the chorionic cytotrophoblasts. The expression of FAAH was the highest in amniotic epithelial cells, chorionic cytotrophoblast and maternal decidua layer. Our results suggest that the human placenta is a likely target for cannabinoid action and metabolism. This is consistent with a placental site of action of endocannabinoids and cannabis being responsible, at least in part, for the poor outcomes associated with cannabis consumption and pathology in the endocannabinoid system during pregnancy. PMID- 12744924 TI - Subset classification of mouse uterine natural killer cells by DBA lectin reactivity. AB - Uterine Natural Killer (uNK) cells are a transient lymphocyte population found in the pregnant uteri of human and rodents. The pregnant uterine environment appears to influence migration, differentiation and suppression of the cytolytic activation of uNK cells but the mechanisms involved in these processes are not well understood. Similarities to circulating NK (cNK) cells are limited. The present study sought to discrimate uNK cells from cNK cells in mice by identification of a unique uNK cell marker. Dolichos biflorus (DBA) lectin, which has high selectivity for glycoconjugates containing N-acetyl D-galactosomine in the terminal position, reacted with the plasma membranes of mouse uNK cells. DBA lectin did not react with other uterine lymphocytes or with cNK cell surfaces in Swiss, CBA-J, C57BL/6, SJL, BALB/c, DBA-2 mice strains. DBA lectin staining was useful for both light and electron microscopy and distinguished 4 uNK cell subtypes that appear to be stages of differentiation. Quantitative evaluation of these 4 uNK cell subtypes over early to late gestational times showed dynamic changes between immature and mature forms in different compartments of the implantation sites and indicated the occurrence of microdomains in the uterus capable of controlling uNK cell proliferation and differentiation. This is the first report showing mouse uNK cells expressing specific molecules not found in other NK cells. Use of this reagent should enhance studies of earlier, non granulated forms of uNK cells and provide new strategies for purification of mouse uNK cells for functional and molecular studies. PMID- 12744925 TI - Glycosylation of the materno-foetal interface in the pregnant viviparous placentotrophic lizard Chalcides chalcides: a lectin histochemical study. AB - Glycosylation of the foeto-maternal interface of the skink Chalcides chalcides has been examined at various stages of gestation using lectin histochemistry. Specimens of incubatory chamber or placenta from early, mid-, late- and near-term pregnancy were fixed and embedded in epoxy resin. Areas of foeto-maternal apposition were probed with a panel of biotinylated lectins followed by an avidin peroxidase revealing system to identify various classes of glycan at the interface. Both the external epithelium of unspecialized bilaminar omphalopleure, which forms by early pregnancy, and chorioallantoic membrane which develops by mid-pregnancy, were composed of two phenotypes, one of which secreted a wide range of glycans including high mannose and complex N-glycan, N-acetyl glucosamine, lactosamine and galactosamine, which became less prominent from mid pregnancy onwards. The uterine epithelium also contained a well-developed secretory apparatus producing a similar range of glycans and there were indications that glycosylated secretions were taken up by the overlying chorioallantois. Foetal vasculature was well developed while maternal vessels appeared more contracted, and both were richly sialylated like their therian equivalents. Our findings indicate that this reptile has evolved a true epitheliochorial placenta with many aspects in common with its therian counterparts but also with unique features of its own. PMID- 12744926 TI - Fibrin-type fibrinoid in placentae from pregnancies associated with maternal smoking: association with villous trophoblast and impact on intervillous porosity. AB - Smoking during pregnancy perturbs maternal haemostasis via activated coagulation which could include greater coagulation (fibrin-type fibrinoid deposition) in the placental intervillous space. This might affect intervillous haemodynamics and transport of oxygen and nutrients to the fetus. Fibrin deposits could influence the sizes and numbers of intervillous spaces ('pores') and perivillous fibrin could reflect changes in the nature or activity of trophoblast. Here, we test whether or not smoking is associated with differences in the composition of villous trophoblast, the amounts and patterns of fibrin and, hence, the dimensions and numbers of intervillous pores. Random samples of placentae were taken from pregnancies classified according to smoking status (non-smokers, light smokers, heavy smokers). Stereology was used to estimate volumes of intervillous space and fibrin, test for differences in trophoblast composition and patterns of fibrin deposition at the villous surface, and determine the impact of deposits on the mean volumes and theoretical numbers of intervillous pores. No group differences were found in total volumes or surfaces of trophoblast or total volume of intervillous fibrin. However, the total surfaces of syncytial knots declined in smokers and the surfaces of syncytial bridges increased. Particularly in heavy smokers, this was associated with reduced deposits of perivillous fibrin at syncytial knots. In all placentae, the greatest deposits occurred where there was trophoblast denudation. Little fibrin was seen on thin regions of syncytium. Regardless of smoking status, intervillous fibrin reduced intervillous pore size and increased pore number. However, heavy smokers had larger pores. Reductions in syncytial knots are consistent with reports that smoking reduces the incidence of trophoblast apoptosis whilst increases in syncytial bridges are consistent with enhanced branching angiogenesis. Results confirm that perivillous fibrin accumulates preferentially at denudation sites. They also suggest that smoking perturbs the normal pattern of fibrin deposition, that the impact is greater in heavy smokers and that the placental site is privileged or active in terms of fibrinolytic or anti-coagulatory activity. This activity seems to reside in thin regions of syncytium. PMID- 12744927 TI - Histomorphology of the placenta and the placental bed of growth restricted foetuses and correlation with the Doppler velocimetries of the uterine and umbilical arteries. AB - The aim of the present study was to evaluate the histomorphology of the placenta and the placental bed and to correlate this with the Doppler study of the uterine and umbilical arteries of intrauterine growth restricted pregnancies. The study group consisted of 47 women with intrauterine growth restricted foetuses. Twenty five uneventful pregnancies with appropriate for gestational age foetuses were selected as controls. Doppler studies of umbilical and uterine arteries were performed within the last week before delivery. Placental bed biopsies were obtained at Caesarean section with direct visualization of the placental site. The incidence of pathologic bed biopsies in control, IUGR with normal uterine artery Doppler velocimetry and IUGR with abnormal uterine artery Doppler velocimetry was 0 per cent, 16.6 per cent and 79.3 per cent respectively (P< 0.001). Placentae from IUGR cases with abnormal umbilical artery Doppler velocimetries had a significantly increased number of villous infarcts, cytotrophoblast proliferation and thickening of the villous trophoblastic basal membrane (P=0.001, P=0.038 and P=0.02 respectively). Abnormal placental bed biopsy pathology was significantly associated with abnormal uterine artery velocimetry (OR 33.7, 6.5-173.6; P< 0.001). Abnormal placental pathology was significantly associated with abnormal umbilical artery Doppler velocimetry (OR 21.04, 3.8-115.9;P< 0.001). Women with both abnormal uterine and umbilical artery Doppler velocimetries were delivered earlier and their babies had lower mean birth and placental weight (P< 0.001). In conclusion, placental bed biopsy and placental pathologies are best reflected by abnormal uterine and umbilical artery velocity waveforms, respectively. The most severe clinical outcomes and perinatal mortality are present when both uterine and umbilical districts are altered. PMID- 12744928 TI - Intralobular differences in antioxidant enzyme expression and activity reflect the pattern of maternal arterial bloodflow within the human placenta. AB - The aim was to determine whether the activities and mRNA concentrations of antioxidant enzymes in human placental tissues reflect the prevailing oxygen tension or developmental maturity of the villi. Advantage was taken of contrasting gradients within lobules of the mature placenta. The central region is well-oxygenated compared to the periphery, owing to the direction of maternal blood flow. However, central villi are morphologically and enzymatically immature compared to peripheral villi. Activity of catalase (t=8.72, P< 0.001) and glutathione peroxidase (t=2.17,P< 0.05) was higher in central than peripheral villi, but no difference was detected for total superoxide dismutase (t=1.08, P> 0.05). The degree of change in catalase activity across the lobule correlated closely with the radius (r=-0.70, P< 0.01). The mRNA concentration was higher in the centre for catalase (t=2.81, P< 0.05) and for glutathione peroxidase (t=3.33, P< 0.05), but no differences were found for copper/zinc or manganese superoxide dismutase. In separate experiments, first trimester villi cultured under 10 per cent oxygen contained higher concentrations of catalase mRNA than controls maintained under 2.5 per cent oxygen. We conclude that the activities of catalase and glutathione peroxidase reflect gradients established by the pattern of maternal intralobular bloodflow, and that oxygen tension is one regulatory factor in vitro. PMID- 12744929 TI - Maternal diabetes mellitus is associated with altered deposition of fibrin-type fibrinoid at the villous surface in term placentae. AB - Placentae from control and diabetic patients were used to test three null hypothesis: (1) there are no significant group differences in the volumes of villous syncytiotrophoblast compartments or intervillous fibrin-type fibrinoid, (2) perivillous fibrin-type fibrinoid is deposited randomly at the surface of trophoblast, and (3) amounts and deposition patterns of perivillous fibrin-type fibrinoid do not vary between groups. Term placentae were collected from non diabetic subjects and five groups of diabetic women classified according to duration, severity and insulin dependence. Tissue specimens and sections were obtained by uniform random sampling. Volumes and surface areas of fibrin-type fibrinoid and trophoblast compartments (thin, syncytial knot, syncytial bridge and denuded regions) were estimated stereologically and compared using variance, chi-squared and contingency table analyses. As to null hypothesis (1), no group differences in volumes of trophoblast compartments were found but volumes of intervillous fibrin-type fibrinoid were greater in the non-insulin-dependent diabetic group. As to null hypothesis (2), regardless of group, fibrin-type fibrinoid was deposited preferentially at sites of denudation in every placenta examined. As to null hypothesis (3), villous surface areas occupied by perivillous fibrin-type fibrinoid were greater in type 1 (insulin-dependent) diabetics with complications (diabetic nephropathy or retinopathy). The surfaces of trophoblast occupied by fibrin-type fibrinoid were also notably larger in non insulin-dependent diabetics and type 1 diabetics with complications. Except for the surface of denudation sites (which also increased in diabetes), there were no differences in the surfaces of trophoblast regions. These results confirm that the haemostatic steady state is perturbed in the diabetic placenta, that perivillous fibrin-type fibrinoid is deposited preferentially at sites of epithelial loss/damage, and that some diabetic groups are affected differentially. PMID- 12744930 TI - Expression of pregnancy-associated plasma protein-A (PAPP-A) during human villous trophoblast differentiation in vitro. AB - Pregnancy-associated placental protein-A (PAPP-A), first isolated from maternal serum, has been identified as a metalloprotease cleaving insulin-like growth factor binding protein-4 (IGFBP-4). The source of PAPP-A during pregnancy is unclear. We therefore investigated PAPP-A expression during in vitro human villous cytotrophoblast cell (CT) differentiation into syncytiotrophoblast (ST). CT were isolated from normal first trimester, second trimester and term placentae (n=10) and cultured to form ST. PAPP-A mRNA was quantified by real-time PCR, and PAPP-A protein expression was studied by immunocytochemistry and TRACE technology with specific monoclonal antibodies. PAPP-A mRNA expression in total placental extracts increased during the course of pregnancy. PAPP-A protein was detected in the cytoplasm of both CT and ST. ST formation in vitro was associated with a 19 fold increase in PAPP-A mRNA expression and an 8-fold increase in PAPP-A secretion into the culture medium. No significant difference in PAPP-A production was observed between cultured cells isolated from early and term placentae. In conclusion, PAPP-A production in vitro, is associated to the differentiation of villous cytotrophoblast cells into syncytiotrophoblast, independently of the age of gestation. PMID- 12744931 TI - Pre-eclampsia and maternal anaemia display reduced apoptosis and opposite invasive phenotypes of extravillous trophoblast. AB - During pregnancy extravillous trophoblast invades maternal uterine tissues and remodels spiral arteries. Maternal anaemia and early onset pre-eclampsia are associated with perturbed trophoblast biology. We systematically compared numerical density, invasive depth and apoptosis rates of extravillous trophoblast in uterine tissues taken from hysterectomies following Caesarean section after normal pregnancies (n=4) or pregnancies complicated by pre-eclampsia (n=5) or anaemia (n=6). Full thickness sections of the placental bed were studied by immunohistochemistry using anti-active caspase 3, anti-cytokeratin 7, anti-lamin B, M30, Mib-1, anti-PARP, and by the TUNEL assay. In normal pregnancy extravillous trophoblast invaded 2.04+/-0.19 mm (mean+/-SEM ) from the endometrial-myometrial border into the myometrium; in pre-eclampsia 0.67+/-0.14 mm (P< 0.01), and in anaemia 3.84+/-0.21 mm (P< 0.001). The endometrial trophoblast density in normal pregnancy was 2.44+/-0.37 cells per 60,000 microm(3), in pre-eclampsia was 1.04+/-0.15 (P< 0.01), and in anaemia was 3.10+/ 0.32. The rate of apoptotic extravillous trophoblast (M30-positive) in the endometrium in normal pregnancy was 7.17+/-1.46 per cent, in pre-eclampsia 4.4+/ 0.71, and in anaemia 2.1+/-0.42 (P< 0.01). Maternal anaemia leads to general tissue hypoxia throughout gestation. Increased invasive depth could be explained by hypoxia-stimulated mitosis and decreased apoptosis of extravillous trophoblast. Reduced trophoblast invasion in pre-eclampsia cannot be explained by higher rates of apoptosis. PMID- 12744932 TI - Vascular endothelial growth factor is a chemoattractant for trophoblast cells. AB - A role for angiogenic growth factors in trophoblast invasion has been postulated. Directional motility (chemotaxis) is an important function of trophoblast cells. We have previously shown that vascular endothelial growth factor (VEGF) increases the random movement of trophoblast cells although placental growth factor (PlGF) has no effect. Heparin inhibited this effect of VEGF. Motility of trophoblast cells has been proposed to be mediated by a nitric oxide (NO) pathway. We hypothesized that VEGF but not PlGF would be chemotactic for trophoblast cells. Chemotaxis of a first trimester extravillous trophoblast cell line, SGHPL-4, and primary isolates of first trimester and term trophoblast cells was measured using a Boyden chamber. Initial experiments to optimize the time of the experiment and identify a positive control were performed. Subsequent experiments ran for 20 h, used 0.5 per cent FBS or 10 ng/ml PDGF as negative and positive controls and were performed in triplicate. VEGF (1, 10 and 100 ng/ml+/-1 microg/ml heparin or +/ 100 microM L-NAME) and PlGF (1, 10, 100 ng/ml) were tested. The chamber was placed in a 5 per cent CO(2) in air, 37 degrees C incubator. The number of cells in the lower chamber were counted. There was a dose dependent increase in chemotactic motility of the SGHPL-4 cell line and term trophoblast cells in response to VEGF. PlGF had no effect on the movement of the first trimester trophoblast cell line but did increase the motility of the term trophoblast cells in a dose dependent manner. Heparin increased the cellular motility of both cell types alone. It also further enhanced the chemoactivity of VEGF on the term trophoblast cells but not the cell line. L-NAME did not affect the VEGF stimulated motility of the first trimester cell line. However, in the term trophoblast cells L-NAME increased the directional cellular motility in the absence of, or in the presence of low concentrations of VEGF. In conclusion, the first trimester and term trophoblast cells appeared to respond differently to the various factors tested in the present study that may reflect differential cellular function as gestation progresses. PMID- 12744933 TI - Correlative microscopy of ultrathin cryosections is a powerful tool for placental research. AB - In this report, we describe procedures for correlative fluorescence and electron microscopy in immunocytochemical studies on the human placenta. Ultrathin cryosections of placenta were used for detection of the distribution of antigens by immunofluorescence and subsequently by immunoelectron microscopy of the same ultrathin cryosection. This methodology has certain advantages over conventional immunohistochemistry and immunoelectron microscopy. The advantages are, most notably, that the same exact structures are examined by both imaging modalities. In addition, since the tissue is physically sectioned (50-100 nm thickness), greater resolution for fluorescence can be obtained in the z-dimension than can be obtained by optical sectioning in confocal microscopy. This last point is of particular importance for discriminating between structures closely stacked in the z-dimension. In this report, we have determined the distribution of caveolin 1 in ultrathin cryosections of terminal villi of the human term placenta. We demonstrate that the use of ultrathin cryosections is a powerful approach for immunofluorescence and correlative microscopy for the in situ localization of antigens. PMID- 12744934 TI - GLUT12 expression in human placenta in first trimester and term. AB - The aim of this study was to characterize the expression of a novel glucose transporter protein GLUT12 in human placenta. GLUT12 mRNA expression was identified by RT-PCR in extracts from five normal term placentae and in extracts from cultured cells of the JAR, JEG-3 and HTR-8Svneo cell lines. In further studies, paraffin sections of first trimester tissue from chorionic villus sampling and term tissue obtained after delivery were analysed by immunohistology with a GLUT12 specific polyclonal antibody. GLUT12 immunoreactivity was expressed predominantly in the syncytiotrophoblast and in extra-villous trophoblast cells in first trimester tissues at 10, 11 and 12 weeks' gestation. In term tissue, however, GLUT12 staining was not detected in syncytiotrophoblast and was found predominantly in villous vascular smooth muscle cells and villous stromal cells. These results suggest that there is a dynamic spatial and temporal expression pattern for the novel glucose transporter GLUT12 in human placenta. PMID- 12744935 TI - Evidence of association of the ratio birth weight/placental weight with genetic factors located in the short arm of chromosome 1. PMID- 12744936 TI - Profiling genes expressed in human fetal cartilage using 13,155 expressed sequence tags. AB - OBJECTIVE: To analyze the gene expression profile of human fetal cartilage by expressed sequence tags (ESTs). METHODS: A human fetal cartilage (8-12 weeks) cDNA library was constructed using the lambda ZAP Express vector. ESTs were obtained by partial sequencing of cDNA clones. The basic local alignment search tool algorithm was used to compare all generated ESTs to known sequences. RESULTS: A total of 13,155 ESTs were analyzed, of which 8696 ESTs (66.1%) matched known genes, 53 ESTs (0.4%) were putatively novel (with no match) and the rest matched other ESTs, genomic DNA and repetitive sequences. Importantly, we identified 2448 unique known genes through non-redundancy analysis of the known gene matches, which were then functionally categorized. The tissue specificity of this library was reflected by its EST profile of the extracellular matrix (ECM) proteins. Collagens were the major transcripts, representing 68.5% of the ECM proteins. Proteoglycans were the second most abundant, constituting 9.5%. Collagen type II was the most abundant gene of all. Glypican 3, decorin and aggrecan were the major transcripts of proteoglycans. Many genes involved in cartilage development were identified, such as insulin-like growth factor-II, its receptor and binding proteins, connective tissue growth factor and fibroblast growth factors. Proteases and their regulatory factors were also identified, including matrix metalloprotease 2 and tissue inhibitor of metalloproteinase 1. CONCLUSIONS: The EST approach is an effective way of characterizing the genes expressed in cartilage. These data represent the most extensive molecular information on human fetal cartilage to date. The availability of this information will serve as a basis for further research to identify genes that are essential in cartilage development. PMID- 12744938 TI - The radiographic joint space width in clinically normal hips: effects of age, gender and physical parameters. AB - OBJECTIVE: The aim of the study was to assess the relationship of the radiographic joint space width (JSW) in the hip with age and a variety of physical parameters in a clinically non-arthritic population in order to identify potential age-related changes. DESIGN: One hundred and eighteen patients (58F/60M, age range 20-79 years) who underwent supine abdominal radiography for non-rheumatological indications and had no hip pain were evaluated. Height, weight and leg lengths were measured. JSW was quantified manually by a dial caliper, and femoral head diameters were determined for each hip. RESULTS: Overall, JSW was 3.61mm+/-0.58 (mean+/-SD) in the right (R) and 3.63mm+/-0.59 in the left (L) hip (range 2.34-6.1mm). There was no age-related decline in the JSW, either by decade (P=0.5 and 0.6, for R and L hips, respectively), or by individual age (Spearman's rho=-0.108 and 0.057, P=0.3 and 0.5 for R and L hips, respectively); similarly, no age-related changes were observed when each gender was analyzed separately. Women had significantly narrower hip JSW than men (P=0.001 and 0.01, R and L hips, respectively). However, gender was no longer significant after height was taken into account (P=0.26 and 0.45, for R and L hips, respectively). JSW correlated significantly with height, weight (which also correlated with height) (r=0.31/0.27 and 0.29/0.28 for height and weight R/L, respectively, P<0.004 for each), but not with body mass index (P=0.62 and 0.57, R and L, respectively). Hip JSW significantly correlated also with femoral head diameter and leg length (r=0.38/0.29 and 0.25/0.19 for femoral head and leg length, R/L, respectively). CONCLUSIONS: No effect of aging was detected on the radiographic JSW of the hip among normal individuals even at advanced ages. In contrast, height, femoral head diameter and leg length were directly related to JSW. PMID- 12744937 TI - Functional barrier principle for growth-factor-based articular cartilage repair. AB - OBJECTIVE: Induction of growth-factor-based repair in full-thickness articular cartilage defects can be impaired by the upgrowth of blood vessels and new bone into the cartilaginous compartment. We postulated that if an antiangiogenic factor (suramin) is included in the chondrogenic matrix applied to the cartilaginous compartment of a full-thickness defect, vascular upgrowth and therefore bone formation will be inhibited (functional barrier principle). DESIGN: Full-thickness defects were created in miniature pigs and the bony portion filled with a chondrogenic matrix. The cartilaginous compartment was filled with the same matrix which additionally contained suramin, either in a free form or in free and liposome-encapsulated forms. Animals were sacrificed 8 weeks after surgery and the extent to which bone tissue had encroached on the cartilaginous compartment was graded semiquantitatively using light microscopy. RESULTS: In 63% of the control defects, bone represented more than 50% of the repair tissue present. In 10% of the defects treated with free suramin, bone upgrowth was completely inhibited; in 55%, osseous tissue occupied 1-10% of the cartilaginous space and in the other 35%, it represented 11-50% of the repair tissue present in this compartment. In 69% of the defects treated with free and liposome-encapsulated suramin, bone upgrowth into the cartilaginous compartment was completely inhibited; in the remaining 31%, osseous tissue occupied no more than 1-10% of this space. CONCLUSIONS: To be effective, an antiangiogenic factor needs to be present at a sustained level throughout the chondrogenic treatment course. PMID- 12744939 TI - Glucosamine and chondroitin sulfate: biological response modifiers of chondrocytes under simulated conditions of joint stress. AB - OBJECTIVE: To test the hypothesis that chondrocytes are more responsive to the chondroprotective agents, glucosamine (glcN) and chondroitin sulfate (CS), under in vitro conditions simulating in vivo joint stress. DESIGN: Synthetic and anticatabolic activities of bovine articular cartilage were assayed using 35 sulfate labeling and assaying the specific activity of glycosaminoglycans (GAGs) under the conditions of enzyme-induced matrix depletion, heat stress, mechanical compression and cytokine stress. RESULTS: The response of cartilage to simulated conditions of in vivo stress varies, depending on the type stress and age of the animal. Cartilage from aged animals was more responsive to stress and to glcN and CS. Pronase-induced matrix depletion and mechanical stress increased proteoglycan synthetic activity. Exposure to glcN and CS significantly enhanced this stress response from 85 to 191% and from 40 to 1000%, respectively. Heat stress and stromelysin digestion decreased synthetic activity, which was reversed or normalized on exposure to glcN and CS. Cartilage from young joints was somewhat refractory to the level of stress imposed and to treatment with glcN and CS. CONCLUSION: The metabolic response of cartilage from aged animals to glcN and CS under simulated conditions of in vivo stress is significantly greater than that seen in nonstressed or young tissue. By enhancing the "protective" metabolic response of chondrocytes to stress, glcN and CS may improve its ability for repair and regeneration. These observations suggest that these compounds function as biological response modifiers (BRMs), agents which boost natural protective responses of tissues under adverse environmental conditions. PMID- 12744940 TI - Proteoglycan metabolism and viability of articular cartilage explants as modulated by the frequency of intermittent loading. AB - OBJECTIVE: This study was designed to systematically determine whether and to what extent the frequency of intermittent loading modulates the biosynthesis and release of proteoglycans (PGs), and to assess chondrocyte viability within mature bovine articular cartilage explants exposed to different loading patterns. METHODS: Cultured full-thickness cartilage explants from the weight-bearing area of healthy bovine fetlock joints were exposed to intermittently applied, uniaxial cyclic loads by introducing a sinusoidal waveform of 0.1, 0.5 or 1.0Hz, frequency and a peak stress of 0.5MPa for a period of 6 days. The cyclic loads were applied for 5, 10 or 20s followed by a period of unloading lasting 10, 100 or 1000s. The incorporation of radiolabeled sulfate into glycosaminoglycans (GAGs) during the final 18h, the content of GAGs and DNA, the deformation of loaded explants as well as the viability of chondrocytes within the different zones of explants were determined. RESULTS: PG synthesis and loss of endogenous PGs were non-linearly and independently regulated by the frequency of the chosen intermittent load, whereas the release of newly synthesized PGs remained unaffected. The viability of chondrocytes within the superficial zone decreased drastically under intermittent loading in a manner independent of the frequency applied. CONCLUSIONS: Our results confirm the hypothesis that the frequency of intermittent loading is an important mechanical factor controlling the metabolic activities of chondrocytes. They also implicate that an initially healthy cartilage explant can be mechanically manipulated to generate an in vitro model of degenerative, osteoarthritic-like cartilage. PMID- 12744941 TI - Reliability of a quantification imaging system using magnetic resonance images to measure cartilage thickness and volume in human normal and osteoarthritic knees. AB - OBJECTIVE: The aim of this study was to evaluate the reliability of a software tool that assesses knee cartilage volumes using magnetic resonance (MR) images. The objectives were to assess measurement reliability by: (1) determining the differences between readings of the same image made by the same reader 2 weeks apart (test-retest reliability), (2) determining the differences between the readings of the same image made by different readers (between-reader agreement), and (3) determining the differences between the cartilage volume readings obtained from two MR images of the same knee image acquired a few hours apart (patient positioning reliability). METHODS: Forty-eight MR examinations of the knee from normal subjects, patients with different stages of symptomatic knee osteoarthritis (OA), and a subset of duplicate images were independently and blindly quantified by three readers using the imaging system. The following cartilage areas were analyzed to compute volumes: global cartilage, medial and lateral compartments, and medial and lateral femoral condyles. RESULTS: Between reader agreement of measurements was excellent, as shown by intra-class correlation (ICC) coefficients ranging from 0.958 to 0.997 for global cartilage (P<0.0001), 0.974 to 0.998 for the compartments (P<0.0001), and 0.943 to 0.999 for the condyles(P<0.0001). Test-retest reliability of within-reader data was also excellent, with Pearson correlation coefficients ranging from 0.978 to 0.999 (P<0.0001). Patient positioning reliability was also excellent, with Pearson correlation coefficients ranging from 0.978 to 0.999 (P<0.0001). CONCLUSIONS: The results of this study establish the reliability of this MR imaging system. Test retest reliability, between-reader agreement, and patient positioning reliability were all extremely high. This study represents a first step in the overall validation of an imaging system designed to follow progression of human knee OA. PMID- 12744942 TI - Assessment of progression in knee osteoarthritis: results of a 1 year study comparing arthroscopy and MRI. AB - OBJECTIVE: The objectives of this study were to determine the sensitivity to change of magnetic resonance imaging (MRI) quantification of chondropathy after 1 year in osteoarthritis of the medial tibiofemoral compartment and to assess the predictive value of subchondral bone marrow edema and bone abnormalities on progression of chondropathy. DESIGN: Twenty patients with symptomatic knee osteoarthritis of the medial compartment underwent a prospective, longitudinal study. All patients were evaluated the same day at entry and after 1 year by plain weight-bearing radiographs, MRI with a three-dimensional gradient-echo sequence, using a 0.2-T dedicated MR unit, and arthroscopy. The medial tibiofemoral chondropathy was quantified blindly with MRI and arthroscopy using the French Society of Arthroscopy (SFA) score. Presence of subchondral bone marrow edema and bone abnormalities on initial MRI was recorded in order to evaluate their influence on both unchanged and worsened chondropathy after 1 year. RESULTS: After 1 year, no statistically significant changes were observed with plain radiographs and arthroscopy. At variance, a statistically significant worsening of chondropathy was found with MRI using the SFA-MR score (P=0.01). SFA MR score was the most responsive outcome. Absence of subchondral bone abnormalities and bone marrow edema on initial MR assessment predicted absence of worsening of chondropathy after 1 year. CONCLUSION: MRI appears promising for evaluating progression of knee osteoarthritis. PMID- 12744943 TI - Joint space width of the tibiofemoral and of the patellofemoral joint in chronic knee pain with or without radiographic osteoarthritis: a 2-year follow-up. AB - OBJECTIVES: To assess the interval change of the minimal joint space width (MJS) in radiographs of the tibiofemoral (TF) joint and of the patellofemoral (PF) joint with a 2-year follow-up in middle-aged people with longstanding knee pain with or without radiographic osteoarthritis (OA) and to study the precision of the MJS measurements. DESIGN: In the format of a prospective study of early OA the signal knee in 55 people, 28 men and 27 women (aged 41-57 years, median 50), with chronic knee pain at inclusion was examined with a 2-year interval (median 25 months, range 21-30). The MJS of the TF joint was measured using a flexed PA view in weightbearing and the MJS of the PF joint using an axial view in standing. RESULTS: The MJS of the TF joint decreased medially by 0.056+/-0.44mm (n.s.) and increased laterally by 0.080+/-0.51mm (n.s.) during the time of observation. In knees with an MJS medially that was less or the same as compared with the lateral compartment, the MJS decreased by 0.14+/-0.38mm (p=0.038) and in a subgroup of these knees, without osteophytes, the MJS decreased by 0.14+/ 0.27mm (p=0.018). The MJS of the PF joint decreased by 0.019mm (n.s.) during the time of observation. The coefficient of variation for intra- and interobserver MJS measurements of the TF joint was 1.0 and 1.1% medially and 2.3 and 2.7% laterally, and for measurement error 6.9% medially and 4.8% laterally, respectively. The coefficient of variation for intra- and interobserver MJS measurements of the PF joint was 8.1 and 5.8% medially and 7.5 and 10.1% laterally and for the measurement error it was 8.1% medially and 8.5% laterally, respectively. CONCLUSIONS: A statistically significant reduction of the MJS was only demonstrated in the medial compartment of the TF joint in those individuals who had an MJS in this compartment which was less or the same as compared with the lateral compartment as well as in a subgroup of these knees without osteophytes. The radiographic examinations and the MJS measurements were reproducible. PMID- 12744944 TI - Response to Torrance et al. [Osteoarthritis and Cartilage 2002; 10:518-27]. PMID- 12744946 TI - Attentional resource and processing speed limitations during sentence processing in Parkinson's disease. AB - Several studies have suggested that patients with Parkinson's disease (PD) have sentence comprehension difficulty in part because of their limited executive resources. However, these assessments confound the executive resources contributing to sentence comprehension with the resources needed for task performance. In the present study, we used a word detection technique that minimizes task demands in order to evaluate attentional and processing speed resources during the comprehension of simple sentences without subordinate clauses and sentences containing subject-relative and object-relative center embedded subordinate clauses. We found that PD patients have poor sensitivity to phonetic errors embedded in unbound grammatical morphemes, regardless of the clausal structure of the sentence, suggesting difficulty attending to grammatical morphemes. We also found that PD patients are significantly slowed in their sensitivity to phonetic errors in content words embedded in object-relative center-embedded sentences. Slowed sensitivity to content words in object-relative sentences was correlated with timed executive measures of planning. On a traditional measure of comprehension, these PD patients were impaired for sentences containing object-relative center-embedded clauses compared to sentences with subject-relative center-embedded clauses, and comprehension of object-relative sentences was correlated with executive measures. Our findings are consistent with the claim that limited executive resources for strategic attention and processing speed contribute to the sentence comprehension difficulties of PD patients. PMID- 12744947 TI - A fMRI study of word retrieval in aphasia. AB - The neural mechanisms underlying recovery of cognitive functions are incompletely understood. Aim of this study was to assess, using functional magnetic resonance (fMRI), the pattern of brain activity during covert word retrieval to letter and semantic cues in five aphasic patients after stroke, in order to assess the modifications of brain function which may be related to recovery. Four out of five patients had undergone language recovery, according to standard testing, after at least 6 months of rehabilitation. The cerebral activation of each patient was evaluated and compared with the activation pattern of normal controls studied with the same fMRI paradigm. In the patients, the pattern of brain activation was influenced by the site and extent of the lesion, by the degree of recovery of language, as reflected by task performance outside the scanner, and by task requirements. In the case of word retrieval to letter cues, a good performance was directly related to the activation in Broca's area, or in the right-sided homologue. On the other hand, in the case of semantic fluency, the relationship between performance level and activation was less clear-cut, because of extensive recruitment of frontal areas in patients with defective performance. These findings suggest that the performance in letter fluency is dependent on the integrity of the left inferior frontal cortex, with the participation of the homologous right hemispheric region when the left inferior frontal cortex is entirely of partially damaged. Semantic fluency, which engages the distributed network of semantic memory, is also associated with more extensive patterns of cerebral activation, which however appear to reflect retrieval effort rather than retrieval success. PMID- 12744948 TI - fMRI of developmental stuttering: a pilot study. AB - The purpose of this investigation was to explore the feasibility of fMRI in the study of developmental stuttering. Speech contrasts (loud versus silent reading) and language contrasts (reading of semantically meaningful text versus nonsense words) of six developmental stutterers and six nonstutterers were compared using a commercial 1 Tesla MR-Scanner (Siemens Expert). Results indicate that mapping cortical function in persons who stutter is indeed feasible, even with a 1TMR system. Compared to normals the stutterers seemed to employ different and particularly less differentiated auditory and motor feedback strategies in speech. They apparently rely on auditory processing and on cerebellar contribution as much during silent reading as during reading aloud. Moreover, they showed a greater involvement of the right hemisphere in language processing, activating not only the typical language areas on the left but also and with equal magnitude the right side homologues of these areas. In spite of the promising results, at present several practical problems such as possible movement artifacts and possible masking through scanner noise still hamper a more straightforward use of fMRI in the study of developmental stuttering. PMID- 12744949 TI - Does right hemisphere damaged patients' impaired performance on a sentence insertion task indicate a syntactic or a lexical level deficit? AB - The present paper focuses on two studies, one on English (Schneiderman & Saddy, 1988) and the other on Italian (De Vreese, Neri Rubichi, & Salvioli, 1996), which report on right hemisphere damaged subjects' inability to correctly insert a word or phrase into an already well-formed sentence under certain stimulus conditions. Despite their strikingly similar findings, the results of the studies are interpreted quite differently. Schneiderman & Saddy consider the deficit to be syntactic, while De Vreese et al. view it as occurring at the lexical category or morphological level. An analysis and regrouping of the stimulus items from both studies indicate that a single syntactic interpretation could account for their results. Suggestions are made for further analyses and research to increase our understanding of the syntactic capacities of the intact right hemisphere. PMID- 12744950 TI - Does the right hemisphere take over after damage to Broca's area? the Barlow case of 1877 and its history. AB - In 1877 Thomas Barlow, a London physician, published a remarkable case of functional recovery of speech following brain damage. It involved a 10-year-old boy who had lost his speech, regained it, and lost it again before he died from a disorder that affected his heart and produced embolisms that subsequently affected other organs, including his brain. Examination of the boy's brain revealed two focal regions of softening; one that affected Broca's area and the left facial-motor area, and another, which occurred weeks later, in the homologous regions of the right hemisphere. Although Barlow was most concerned with motor deficits, others at the turn of the century began to cite this case as strong evidence that the corresponding region of the right hemisphere can take over speech functions for Broca's area on the left. Whether this case really provides good support for functional takeover or vicariation theory is critically evaluated in the light of contemporary research, including PET scan studies involving damage to Broca's speech region. PMID- 12744951 TI - The brain generates its own sentence melody: a Gestalt phenomenon in speech perception. AB - Brain processes underlying spoken language comprehension comprise auditory encoding, prosodic analysis and linguistic evaluation. Auditory encoding usually activates both hemispheres while language-specific stages are lateralized: analysis of prosodic cues are right-lateralized while linguistic evaluation is left-lateralized. Here, we investigated to what extent the absence of prosodic information influences lateralization. MEG brain-responses indicated that syntactic violations lead to early bi-lateral brain responses for syntax violations. When the pitch of sentences was flattened to diminish prosodic cues, the brain's syntax response was lateralized to the right hemisphere, indicating that the missing pitch was generated automatically by the brain when it was absent. This represents a Gestalt phenomenon, since we perceive more than is actually presented. PMID- 12744952 TI - Distributed cortical networks for syntax processing: Broca's area as the common denominator. AB - Different types of syntactic information (word category, grammatical gender) are processed at different times during word recognition. However, it is an open issue which brain systems support these processes. In the present event-related fMRI study, subjects performed either a syntactic gender decision task on German nouns (GEN), a word category decision task (WC, nouns vs. prepositions), or a physical baseline task (BASE). Reaction times in WC were faster than in GEN, supporting earlier electrophysiological results. Relative to BASE, both syntactic tasks activated the inferior tip of BA 44. In addition, BA 45 showed activation in GEN, whereas BA 47 was activated in WC. The imaging data indicate that the inferior portion of BA 44 together with type-specific prefrontal areas supports both initial word category related and later syntactic processes. PMID- 12744954 TI - Dyslexia linked to talent: global visual-spatial ability. AB - Dyslexia has long been defined by deficit. Nevertheless, the view that visual spatial talents accompany dyslexia has grown, due to reports of individuals with dyslexia who possess visual-spatial strengths, findings of elevated incidence of dyslexia in certain visual-spatial professions, and the hypothesis that left hemisphere deficits accompany right-hemisphere strengths. Studies have reported superior, inferior, and average levels of visual-spatial abilities associated with dyslexia. In two investigations, we found an association between dyslexia and speed of recognition of impossible figures, a global visual-spatial task. This finding suggests that dyslexia is associated with a particular type of visual-spatial talent-enhanced ability to process visual-spatial information globally (holistically) rather than locally (part by part). PMID- 12744953 TI - Effect of lexical cues on the production of active and passive sentences in Broca's and Wernicke's aphasia. AB - This study compared the sentence production abilities of individuals with Broca's and Wernicke's aphasia in an attempt to explore the extent to which impaired lexical retrieval impedes sentence production. The ability to produce active and passive reversible and non-reversible sentences was examined when varying amounts of lexical information was provided. The results showed that both Wernicke's and Broca's aphasic individuals were impaired in passive sentence production and that these difficulties were not overcome when lexical cues (the relevant nouns and uninflected verb) were provided. However when auxiliary and past tense morphemes were provided along with the verb stem, production of passive sentences improved drastically for both groups. Analysis of error patterns, however, revealed differences between the two groups, suggesting that Broca's aphasic subjects may find passive sentences difficult due to problems with retrieving the relevant grammatical morphemes. Subjects with Wernicke's aphasia may have been unable to automatically access the passive sentence structure. PMID- 12744955 TI - Facilitation and disruption of lateralized syllable processing by unattended stimuli in the opposite visual field. AB - The influence of lateralized unattended stimuli on the processing of attended stimuli in the opposite visual field can shed light on the nature of information that is transferred between hemispheres. On a cued bilateral task, participants tried to identify a syllable in the attended visual field, which elicits a left hemisphere (LH) advantage and different processing strategies by the two hemispheres. The same or a different syllable or a neutral stimulus appeared in the unattended field. Transmission of unattended syllable codes between hemispheres is symmetric, as revealed by equal interference for the two visual fields. The LH is more accurate than the RH in encoding unattended syllables, as indicated by facilitation in the left but not right visual field and a greater frequency of identifiable intrusions into the left than right field. However, asymmetric encoding strategies are different for attended and unattended syllables. PMID- 12744956 TI - Effect of typicality on online category verification of animate category exemplars in aphasia. AB - Normal young, elderly, Broca's aphasic, and Wernicke's aphasic individuals participated in an online category verification task where primes were superordinate category labels while targets were either typical or atypical examples of animate categories or nonmembers belonging to inanimate categories. The reaction time to judge whether the target belonged to the preceding category label was measured. Results indicated that all four groups made significantly greater errors on atypical examples compared to typical examples. Young and elderly individuals, and Broca's aphasic patients performed similarly on the verification task; these groups demonstrated faster reaction times on typical examples than atypical examples. Wernicke's aphasic patients made the most errors on the task and were slowest to respond than any other participant group. Also, these participants were not significantly faster at accepting correct typical examples compared to correct atypical examples. The results from the four groups are discussed with relevance to prototype/family resemblance models of typicality. PMID- 12744957 TI - Weak coherence, no theory of mind, or executive dysfunction? Solving the puzzle of pragmatic language disorders. AB - Deficits in pragmatic language ability are common to a number of clinical populations, for example, right-hemisphere damage (RHD), Autism and traumatic brain injury (TBI). In these individuals the basic structural components of language may be intact, but the ability to use language to engage socially is impaired. Despite the nature of these difficulties being well documented, exactly what causes these difficulties is less clear. Furthermore, the current status of causal explanations for pragmatic difficulties across these populations is divergent and sometimes contradictory. This paper explores the empirical validity of three theories that attempt to explain pragmatic language impairment. It is recommended that a new, more convergent approach to investigating the causes of pragmatic language disability be adopted. PMID- 12744958 TI - Are there differential word length effects in the two visual fields? AB - This study examined whether differential word length effects in the two visual fields imply hemisphere-dependent modes of word recognition. Length was defined as the number of constituent characters of Chinese foreign names (Experiments 1 and 2), as the number of constituent morphemes of three-character words (Experiments 3 and 4), and as that of constituent words of phrases (Experiments 5 and 6). Two types of experimental tasks were adopted, one required linguistic judgments on overall items (Experiments 1, 3, and 5) and the other was target detection tasks performed on the same set of stimuli (Experiments 2, 4, and 6). Five of the six experiments failed to find any kind of interaction between length and visual field. An interaction was observed only for the detection of characters embedded in foreign names, that is, when lexical access is least involved in the task, suggesting that word recognition plays a minimum role in the phenomenon. Other observations suggested that modes of word recognition are more frequency-dependent than hemisphere-dependent, and that Chinese compound words and phrases, although hardly distinguishable, do behave differently. PMID- 12744959 TI - Speed of processing of the visual-orthographic and auditory-phonological systems in adult dyslexics: the contribution of "asynchrony" to word recognition deficits. AB - This study investigated whether "asynchrony" in speed of processing (SOP) between the visual-orthographic and auditory-phonological modalities contributes to word recognition deficits among adult dyslexics. Male university students with a history of diagnosed dyslexia were compared to age-matched normal readers on a variety of experimental measures while event-related potentials and reaction time data were collected. Measures were designed to evaluate auditory and visual processing for non-linguistic (tones and shapes) and linguistic (phonemes and graphemes) low-level stimuli as well as higher-level orthographic and phonological processing (in a lexical decision task). Data indicated that adult dyslexic readers had significantly slower reaction times and longer P300 latencies than control readers in most of the experimental tasks and delayed P200 latencies for the lexical decision task. Moreover, adult dyslexics revealed a systematic SOP gap in P300 latency between the auditory/phonological and visual/orthographic processing measures. Our data support and extend previous work that found SOP asynchrony to be an underlying factor of childhood dyslexia. The present data suggests, however, that among adult dyslexics the between modalities asynchrony occurs at later processing stages than in children. PMID- 12744960 TI - Sensitivity to local sentence context information in lexical ambiguity resolution: evidence from left- and right-hemisphere-damaged individuals. AB - Using a cross-modal semantic priming paradigm, the present study investigated the ability of left-hemisphere-damaged (LHD) nonfluent aphasic, right-hemisphere damaged (RHD) and non-brain-damaged (NBD) control subjects to use local sentence context information to resolve lexically ambiguous words. Critical sentences were manipulated such that they were either unbiased, or biased toward one of two meanings of sentence-final equibiased ambiguous words. Sentence primes were presented auditorily, followed after a short (0 ms) or long (750 ms) interstimulus interval (ISI) by the presentation of a first- or second-meaning related visual target, on which subjects made a lexical decision. At the short ISI, neither patient group appeared to be influenced by context, in sharp contrast to the performance of the NBD control subjects. LHD nonfluent aphasic subjects activated both meanings of ambiguous words regardless of context, whereas RHD subjects activated only the first meaning in unbiased and second meaning biased contexts. At the long ISI, LHD nonfluent aphasic subjects failed to show evidence of activation of either meaning, while RHD individuals activated first meanings in unbiased contexts and contextually appropriate meanings in second-meaning biased contexts. These findings suggest that both left (LH) and right hemisphere (RH) damage lead to deficits in using local contextual information to complete the process of ambiguity resolution. LH damage seems to spare initial access to word meanings, but initially impairs the ability to use context and results in a faster than normal decay of lexical activation. RH damage appears to initially disrupt access to context, resulting in an over reliance on frequency in the activation of ambiguous word meanings. PMID- 12744961 TI - Commentary on A. Beretta et al. 'An ER-fMRI investigation of morphological inflection in German reveals that the brain makes a distinction between regular and irregular forms'. PMID- 12744962 TI - The brain makes a distinction between hard and easy stimuli: comments on Beretta et al. AB - The Beretta et al. study tested an invalid prediction concerning connectionist models of inflectional morphology and the study exhibits a confound between type of stimulus (regular, irregular) and processing difficulty (easy, hard) that invalidates their conclusions. Harder stimuli produced greater activation across a broader range of brain areas, as in previous studies, but the data have no bearing on the rules vs. connections debate. PMID- 12744964 TI - Spectral and temporal degradation of speech as a simulation of morphosyntactic deficits in English and German. AB - Spectral and temporal degradation of the speech stream is increasingly used to model receptive language deficits such as aphasia and developmental language disorders. As with results from patient studies, the specific pattern of receptive deficits can reveal underlying structural and processing characteristics of different languages. Here, we test English- and German speaking college students' auditory comprehension of complex morphosyntactic structures under normal and 'dual-degradation' conditions. The resulting profiles of strength and vulnerability in the two languages highlight the cross-linguistic differences in reliability of syntactic and morphological cues, and closely resemble the deficits observed in previous studies of receptive aphasia. PMID- 12744965 TI - Cognitive neuroscience. PMID- 12744966 TI - Vision and the statistics of the visual environment. AB - It is widely believed that visual systems are optimized for the visual properties of the environment inhabited by the organism. A specific instance of this principle is known as the Efficient Coding Hypothesis, which holds that the purpose of early visual processing is to produce an efficient representation of the incoming visual signal. The theory provides a quantitative link between the statistical properties of the world and the structure of the visual system. As such, specific instances of this theory have been tested experimentally, and have been used to motivate and constrain models for early visual processing. PMID- 12744967 TI - Bayesian models of object perception. AB - The human visual system is the most complex pattern recognition device known. In ways that are yet to be fully understood, the visual cortex arrives at a simple and unambiguous interpretation of data from the retinal image that is useful for the decisions and actions of everyday life. Recent advances in Bayesian models of computer vision and in the measurement and modeling of natural image statistics are providing the tools to test and constrain theories of human object perception. In turn, these theories are having an impact on the interpretation of cortical function. PMID- 12744968 TI - The neural basis of object perception. AB - Humans can recognize an object within a fraction of a second, even if there are no clues about what kind of object it might be. Recent findings have identified functional properties of extrastriate regions in the ventral visual pathway that are involved in the representation and perception of objects and faces. The functional properties of these regions, and the correlation between the activation of these regions and visual recognition, indicate that the lateral and ventral occipito-temporal areas are important in perceiving and recognizing objects and faces. PMID- 12744969 TI - Cortical mechanisms in hearing. AB - Current understanding of neural processing in the auditory cortex has been shaped by a variety of experimental approaches in animals and humans. It remains a daunting challenge to reconcile data as diverse as synaptic properties recorded in a rodent brain slice and functional images of auditory cortex in a behaving human. Nevertheless, the gaps are narrowing through a renewed focus on humans and other primates, a continuing interest in evidence for functional pathways, a broader application of modern imaging techniques, a growing awareness of cortical sensitivity to dynamic features of sounds, and an improved understanding of auditory cortical circuitry. PMID- 12744970 TI - Tracking cognitive processes with functional MRI mental chronometry. AB - Functional magnetic resonance imaging (fMRI) is used widely to determine the spatial layout of brain activation associated with specific cognitive tasks at a spatial scale of millimeters. Recent methodological improvements have made it possible to determine the latency and temporal structure of the activation at a temporal scale of few hundreds of milliseconds. Despite the sluggishness of the hemodynamic response, fMRI can detect a cascade of neural activations - the signature of a sequence of cognitive processes. Decomposing the processing into stages is greatly aided by measuring intermediate responses. By combining event related fMRI and behavioral measurement in experiment and analysis, trial-by trial temporal links can be established between cognition and its neural substrate. PMID- 12744971 TI - Neural correlates of decision processes: neural and mental chronometry. AB - Recent studies aim to explain the duration and variability of behavioral reaction time in terms of neural processes. The time taken to make choices is occupied by at least two processes. Neurons in sensorimotor structures accumulate evidence that leads to alternative categorizations, while other neurons within these structures prepare and initiate overt responses. These distinct stages of stimulus encoding and response preparation support variable but flexible behavior. PMID- 12744972 TI - Cortical mechanisms of space-based and object-based attentional control. AB - Visual attention, the mechanism by which observers select relevant or important information from scenes, can be deployed to locations in space or to spatially invariant object representations. Studies have examined both the modulatory effects of attention on the strength of extrastriate cortical representations, and the control of attention by parietal and frontal cortical circuits. Subregions of parietal and frontal cortex are transiently active when attention is voluntarily shifted between spatial locations or object representations. This transient activity may reflect an abrupt shift in the attentional set of the observer, complementing sustained signals that are thought to maintain a given attentive state. PMID- 12744973 TI - Neural coding of behavioral relevance in parietal cortex. AB - Flexible control of behavior requires the selective processing of task-relevant sensory information and the appropriate linkage of sensory input to action. A great deal of evidence suggests a central role for the parietal cortex in these functions. Recent results from neurophysiological studies in non-human primates and neuroimaging experiments in humans illuminate the importance of parietal cortex for attention, and suggest how parietal neurons might allow the dynamic representation of behaviorally relevant information. PMID- 12744974 TI - Neural correlates of categories and concepts. AB - The ability to readily adapt to novel situations requires something beyond storing specific stimulus-response associations. Instead, many animals can detect basic characteristics of events and store them as generalized classes. Because these representations are abstracted beyond specific details of sensory inputs and motor outputs, they can be easily generalized and adapted to new circumstances. Explorations of neural mechanisms of sensory processing and motor output have progressed to the point where studies can begin to address the neural basis of abstract, categorical representations. Recent studies have revealed their neural correlates in various cortical areas of the non-human primate brain. PMID- 12744975 TI - Basic mechanisms for graded persistent activity: discrete attractors, continuous attractors, and dynamic representations. AB - Persistent neural activity is observed in many systems, and is thought to be a neural substrate for holding memories over time delays of a few seconds. Recent work has addressed two issues. First, how can networks of neurons robustly hold such an active memory? Computer systems obtain significant robustness to noise by approximating analogue quantities with discrete digital representations. In a similar manner, theoretical models of persistent activity in spiking neurons have shown that the most robust and stable way to store the short-term memory of a continuous parameter is to approximate it with a discrete representation. This general idea applies very broadly to mechanisms that range from biochemical networks to single cells and to large circuits of neurons. Second, why is it commonly observed that persistent activity in the cortex can be strongly time varying? This observation is almost ubiquitous, and therefore must be taken into account in our models and our understanding of how short-term memories are held in the cortex. PMID- 12744976 TI - Developmental dyslexia: specific phonological deficit or general sensorimotor dysfunction? AB - Dyslexia research now faces an intriguing paradox. It is becoming increasingly clear that a significant proportion of dyslexics present sensory and/or motor deficits; however, as this 'sensorimotor syndrome' is studied in greater detail, it is also becoming increasingly clear that sensory and motor deficits will ultimately play only a limited role in a causal explanation of specific reading disability. PMID- 12744977 TI - Self-awareness and action. AB - In this review we discuss how we are aware that actions are self-generated. We review behavioural data that suggest that a prediction of the sensory consequences of movement might be used to label actions and their consequences as self-generated. We also describe recent functional neuroimaging experiments and studies of neurological and psychiatric patients, which suggest that the parietal cortex plays a crucial role in the awareness of action. PMID- 12744979 TI - Motor learning and prediction in a variable environment. AB - Traditional studies of motor learning and prediction have focused on how subjects perform a single task. Recent advances have been made in our understanding of motor learning and prediction by investigating the way we learn variable tasks, which change either predictably or unpredictably over time. Similarly, studies have examined how variability in our own movements affects motor learning. PMID- 12744978 TI - Neocortical mechanisms in motor learning. AB - The ability to learn novel motor skills has fundamental importance for adaptive behavior. Neocortical mechanisms support human motor skill learning, from simple practice to adaptation and arbitrary sensory-motor associations. Behavioral and neural manifestations of motor learning evolve in time and involve multiple structures across the neocortex. Modifications of neural properties, synchrony and synaptic efficacy are all related to the development and maintenance of motor skill. PMID- 12744980 TI - Neural population codes. AB - In many regions of the brain, information is represented by patterns of activity occurring over populations of neurons. Understanding the encoding of information in neural population activity is important both for grasping the fundamental computations underlying brain function, and for interpreting signals that may be useful for the control of prosthetic devices. We concentrate on the representation of information in neurons with Poisson spike statistics, in which information is contained in the average spike firing rate. We analyze the properties of population codes in terms of the tuning functions that describe individual neuron behavior. The discussion centers on three computational questions: first, what information is encoded in a population; second, how does the brain compute using populations; and third, when is a population optimal? To answer these questions, we discuss several methods for decoding population activity in an experimental setting. We also discuss how computation can be performed within the brain in networks of interconnected populations. Finally, we examine questions of optimal design of population codes that may help to explain their particular form and the set of variables that are best represented. We show that for population codes based on neurons that have a Poisson distribution of spike probabilities, the behavior and computational properties of the code can be understood in terms of the tuning properties of individual cells. PMID- 12744981 TI - Distinct systems for automatic and cognitively controlled time measurement: evidence from neuroimaging. AB - A recent review of neuroimaging data on time measurement argued that the brain activity seen in association with timing is not influenced by specific characteristics of the task performed. In contrast, we argue that careful analysis of this literature provides evidence for separate neural timing systems associated with opposing task characteristics. The 'automatic' system draws mainly upon motor circuits and the 'cognitively controlled' system depends upon prefrontal and parietal regions. PMID- 12744982 TI - An expanded role for functional neuroimaging in schizophrenia. AB - Functional magnetic resonance imaging is a surprisingly versatile tool in the quest for disentangling the complexities of mental illnesses such as schizophrenia. Yet, the identification of pathognomonic physiological features of the illness or even a consensus regarding the interpretation of reported findings remain unfulfilled goals, in spite of the increasing sophistication of this technology. Nonetheless, by providing quantification of brain function during various cognitive challenges, functional MRI has been used to leap ahead of these quandaries to identify relationships between genetic variation and brain function. By examining recent findings and efforts to link these findings to genes, this article will review these exciting developments in schizophrenia research. PMID- 12744984 TI - Case-based teaching in preventive medicine: rationale, development, and implementation. AB - The importance of prevention teaching is increasingly recognized in medical education, but its implementation in medical school curricula is hampered by its cross-specialty nature, lack of curricular time, and perception as a topic of less importance than the traditional basic and clinical sciences. The Case-Based Series in Population-Oriented Prevention (C-POP) was developed to address national objectives for prevention education in a format that recognizes the students' abilities and preferences for case-based learning. This series uses small-group discussion cases that can be adapted to a variety of settings and instructor capabilities. These cases guide the learners from a specific clinical problem to the broader clinical and population-based prevention issues for the topic. The cases were developed with the use of local health department scenarios and data and have been taught and refined in a number of settings. As part of the curriculum development project, evaluation tools that examined prevention skills and orientation were developed and tested. With its emphasis on small-group learning, clinical relevance, and adaptability to a variety of learner and instructor needs, the C-POP project effectively integrates prevention concepts into medical education. PMID- 12744985 TI - Evaluation of a preventive medicine curriculum: incorporating a case-based approach. AB - BACKGROUND: Evaluating the acquisition of skills in prevention is an increasing priority in prevention education. Assessment instruments were developed to measure student skills before and after an education intervention at State University of New York (SUNY)-Upstate Medical University. METHODS: The evaluation method used three testing instruments that measure preventive medicine skills. We selected three surrogate topics, each their own instrument: sexually transmitted disease, lead toxicity, and ischemic heart disease. All three instruments measure four key preventive medicine skills areas: (1) using and interpreting data sources; (2) measuring disease frequency, including incidence and prevalence; (3) making inferences and identifying bias in data presentations; and (4) identifying appropriate study design and screening tests. Second-year medical students were assessed before and after our preventive medicine course in spring 2002, using our evaluative instruments. RESULTS: Before and after instruction analysis, overall, and by skills area tested revealed a significant increase in student preventive medicine skills (p< or =0.001) in all four categories. On conclusion of the case-based curriculum, students were also asked to rate the cases. The majority (60%) of the students thought the cases were of value to their medical education, and 58% believed that they added to their skills in population prevention. CONCLUSIONS: These instruments can measure change in preventive medicine skills before and after a course in preventive medicine. PMID- 12744986 TI - Measuring orientation to population-based prevention. AB - BACKGROUND: Assessing orientation to population-based preventive alternatives is useful in the evaluation of preventive medicine curricula. METHODS: An instrument was developed using hypothetical situations describing a health issue that could be encountered by a primary care physician. Options to be selected were in three categories: (1) treatment, (2) clinical prevention, or (3) population-based prevention. Respondents allocated 900 points among alternatives for nine situations (100 points per situation). Pilot testing of the instrument was implemented, comparing the responses of public health physicians with two groups (Syracuse and Baltimore) of family medicine physicians. RESULTS: For the overall instrument, the scores on the population scale and population-treatment differential showed statistically significant differences between the public health physicians and both groups of family medicine physicians (p<0.01). For medical students taught the Case-Based Series in Population-Oriented Prevention (C-POP), there was an overall increase in the population scale (p<0.0001) and the population-treatment differential (p<0.0004). CONCLUSIONS: The development of a scenario-based tool to assess changes in population-based prevention orientation shows preliminary promise for future evaluation efforts involving preventive medicine curricula. PMID- 12744987 TI - Integration of the Case-Based Series in Population-Oriented Prevention into a problem-based medical curriculum. AB - This article describes the pilot testing of the Case-Based Series in Population Oriented Prevention (C-POP) and accompanying evaluation instruments in a problem based medical curriculum at the University of Rochester Medical Center. Several of the cases were successfully taught in the curriculum, but the evaluation instruments revealed no change in population-oriented skills or orientation. Issues of implementation and evaluation of the C-POP curriculum are discussed. PMID- 12744988 TI - Preventive medicine teaching cases in the preclinical undergraduate medical curriculum. AB - The implementation of the Case-Based Series in Population-Oriented Prevention (C POP) at the University of California at Davis Medical School is described. The implementation and planning for the integration of a case-based curriculum into a previously didactic-focused preventive medicine course are discussed. Issues include flexibility of the curriculum, student acceptance of case-based learning, and effect of preceptor availability and training on the successful implementation of the C-POP curriculum. PMID- 12744989 TI - Preventive medicine teaching cases for preventive medicine residents. AB - Preventive medicine education is unique in that its successes are measured in groups of people. Conveying this population perspective can be difficult, even to preventive medicine residents, some of whom have been in clinical practice for many years. The Case-Based Series in Population-Oriented Prevention (C-POP) was adapted for use in the New York State Preventive Medicine Residency curriculum. Parts of two of the cases were felt to be too clinical for use in this setting, but the other cases were well received and imparted the desired population perspective. Although the C-POP series was produced for undergraduate medical education, it is generally adaptable to the needs of a preventive medicine curriculum. PMID- 12744990 TI - A user's guide to teaching the C-POP series. PMID- 12744991 TI - Community health assessment. AB - This case-community health assessment-is one of a series of teaching cases in the Case-Based Series in Population-Oriented Prevention (C-POP). It has been developed for use in medical school and residency prevention curricula. The complete set of cases is presented in this supplement to the American Journal of Preventive Medicine. Community health assessment is key to understanding the health problems and priorities of a population. This case outlines a process by which the participants can complete a health assessment of a community using indicator-based methods. Students construct a set of health indicators from a variety of domains, evaluate problems, and report on the health priorities for a community. The students relate identified health issues to underlying behavioral risk factors. PMID- 12744992 TI - Outbreak of tuberculosis in a homeless men's shelter. AB - This case-tuberculosis in a homeless men's shelter-is one of a series of teaching cases in the Case-Based Series in Population-Oriented Prevention (C-POP). It has been developed for use in medical school and residency prevention curricula. The complete set of cases is presented in this supplement to the American Journal of Preventive Medicine. Tuberculosis presents a significant public health challenge. In this teaching case, medical students are given information about four cases of active tuberculosis that occurred over a short period of time in residents of a homeless men's shelter. The students then walk through the steps that a local health department takes to identify and screen those individuals at risk for transmission of tuberculosis during an outbreak. During this process, they learn skills in epidemiology (such as defining "epidemic" and distinguishing uses for incidence and prevalence) as well as in population-based prevention of tuberculosis. Finally students discuss health policy as it relates to the control and prevention of tuberculosis. PMID- 12744993 TI - Racial and ethnic disparity in low birth weight in Syracuse, New York. AB - This case-racial and ethnic disparity in low birth weight-is one of a series of teaching cases in the Case-Based Series in Population-Oriented Prevention (C POP). It has been developed for use in medical school and residency prevention curricula. The complete set of cases is presented in this supplement to the American Journal of Preventive Medicine. Low birth weight is a leading cause of infant mortality. Unfortunately, despite declining rates of infant mortality, racial and ethnic disparities in both low birth weight and infant mortality rates persist. In this teaching case, a clinical vignette is used to draw attention to this public health priority in Syracuse, New York. Students learn essential epidemiology skills such as identifying limitations of sources of data and calculating relative risks, using the example of low birth weight. In performing these skills, students also identify etiologies for such disparity. Finally, students discuss interventions that, when implemented, may decrease infant mortality rates. PMID- 12744994 TI - Sexually transmitted disease in adolescents. AB - This case-sexually transmitted diseases in adolescents-is one of a series of teaching cases in the Case-Based Series in Population-Oriented Prevention (C POP). It has been developed for use in medical school and residency prevention curricula. The complete set of cases is presented in this supplement to the American Journal of Preventive Medicine. Sexually transmitted diseases (STDs) are common among adolescents. This case focuses on the population-based prevention of STDs. The case begins with clinical, diagnostic, and laboratory management of an adolescent female diagnosed with gonorrhea. Students then examine how the diagnosis of an STD can be used as an opportunity to provide preventive care and counseling on safer sexual behaviors. Questions addressing HIV testing and counseling are included in this section. Students proceed to explore how the diagnosis and reporting of an STD calls for interaction, such as partner notification, between clinical providers and public health departments. Finally, students are provided with actual information from local focus groups on attitudes and behaviors of high-risk adolescents. At the conclusion of the case, they are asked to interpret the data and formulate strategies to decrease gonorrhea rates in the community. PMID- 12744995 TI - A critical look at prevention: colorectal cancer screening. AB - This case-colorectal cancer screening-is one of a series of teaching cases in the Case-Based Series in Population-Oriented Prevention (C-POP). It has been developed for use in medical school and residency prevention curricula. The complete set of cases is presented in this supplement to the American Journal of Preventive Medicine. This preventive medicine teaching case discusses the concepts of diagnostic test evaluation, prevention, and screening using the example of colorectal cancer screening. Features of the case include a health policy exercise concerning community screening programs and an exercise in clinical prevention decision making. PMID- 12744996 TI - Bicycle helmet effectiveness in preventing injury and death. AB - This case--bicycle helmet effectiveness--is one of a series of teaching cases in the Case-Based Series in Population-Oriented Prevention (C-POP). It has been developed for use in medical school and residency prevention curricula. The complete set of cases is presented in this supplement to the American Journal of Preventive Medicine. This case examines the cost-effectiveness of three interventions to increase utilization of bicycle helmets to avert head injuries in individuals aged 18 years and under in Onondaga Count NY. Students are initially presented with data on head injuries, hospitalization, and death related to bicycle use. They then appraise a published study on the effectiveness of bicycle helmets in averting head injury. Finally, students work in groups to determine the cost-effectiveness of each intervention by calculating implementation costs and the specific number of head injuries averted associated with intervention. The three interventions are legislative, school, and community based campaigns to increase helmet use. Students are provided with budget estimates and assumptions needed to complete the exercise. Cost-effectiveness analysis, cost-benefit analysis, and related concepts are discussed, including provider versus societal perspectives and importance of sensitivity analysis. PMID- 12744997 TI - Adolescent suicide prevention. AB - This case-prevention of adolescent suicide-is one of a series of teaching cases in the Case-Based Series in Population-Oriented Prevention (C-POP). It has been developed for use in medical school and residency prevention curricula. The complete set of cases is presented in this supplement to the American Journal of Preventive Medicine. This teaching case examines the issue of prevention of adolescent and young adult suicide both at an individual and at a population or community level, using data from the Onondaga County Health Department. In the first section of the case, students are asked to determine whether five deaths related to falling or jumping at a local shopping mall should be considered to be suicidal deaths. Students then develop skills in the reporting as well as in the epidemiology of adolescent suicidal deaths in Onondaga County. As the case progresses, students analyze the results of a local surveillance study of suicidal attempts and ideation. The case concludes with students evaluating a hypothetical screening study intended to reduce the risk of suicidal death and discussing a research design to examine the effectiveness of this prevention strategy. PMID- 12744998 TI - Maternal mortality. AB - This case-maternal mortality-is one of a series of teaching cases in the Case Based Series in Population-Oriented Prevention (C-POP). It has been developed for use in medical school and residency prevention curricula. The complete set of cases is presented in this supplement to the American Journal of Preventive Medicine. Maternal mortality remains an important public health concern, even though it is a rare event. This teaching module introduces five case reports of maternal death to provide a clinical lead into discussions about data sources such as death certificates and their limitations. The students will also calculate maternal mortality rates and explore racial disparities in this health indicator. Finally, the students will develop intervention strategies to identify and prevent maternal mortality. PMID- 12744999 TI - Population-based prevention: a core competency in medical education. PMID- 12745000 TI - Commentary on the C-POP project. PMID- 12745001 TI - Future applications of case-based teaching in population-based prevention. AB - The Case-Based Series in Population-Oriented Prevention (C-POP) introduced in this supplement to the American Journal of Preventive Medicine provides a set of tools to integrate clinical- and community-based prevention into interactive teaching cases. These cases, which address recommended core competencies in prevention education, have been taught to both medical students and preventive medicine residents. Initial experience with the cases indicates that this method is a promising tool to enhance prevention education in medical schools and primary care residencies. Because prevention education is essential in other fields, such as public health and nursing, extension of this approach to other professional schools is possible. Such extension is feasible because the cases are designed to be adaptable for different levels of education, flexible to be tailored to local situations, and expandable to accommodate changes in the field. In addition, the cases can be made accessible to all educators through a national library and in interactive web-based format. This article describes how the C-POP series can be used to strengthen prevention education for a wide audience of physician and nonphysician learners. PMID- 12745002 TI - Familial retinal arteriolar tortuosity: a review. AB - Familial retinal arteriolar tortuosity is a rare disorder with autosomal dominant inheritance. It is characterized by a pathognomic pattern of progressive, pronounced tortuosity of the arterioles of second and third order in the macular and peripapillary area developing during childhood or early adulthood. The disorder can be complicated by intra- or preretinal hemorrhages that occur spontaneously or during physical exertion. These hemorrhages can be asymptomatic if they do not involve the fovea. Clearing of the hemorrhages without treatment and recovery of full visual acuity is the rule, even if the central macula is involved. In most cases, no other vascular malformations or associated systemic diseases have been found. In some cases, retinal hemorrhages may precede the development of the tortuosity or the tortuosity may remain subtle, even if the patients suffer from recurrent hemorrhages. The ophthalmologist should be aware of familial retinal arteriolar tortuosity as a possible cause for retinal hemorrhages. PMID- 12745003 TI - Age-related macular degeneration: etiology, pathogenesis, and therapeutic strategies. AB - Age-related macular degeneration is the principal cause of registered legal blindness among those aged over 65 in the United States, western Europe, Australia, and Japan. Despite intensive research, the precise etiology of molecular events that underlie age-related macular degeneration is poorly understood. However, investigations on parallel fronts are addressing this prevalent public health problem. Sophisticated biochemical and biophysical techniques have refined our understanding of the pathobiology of drusen, geographic atrophy, and retinal pigment epithelial detachments. Epidemiological identification of risk factors has facilitated an intelligent search for underlying mechanisms and fueled clinical investigation of behavior modification. Gene searches have not only brought us to the cusp of identifying the culpable gene loci in age-related macular degeneration, but also localized genes responsible for other macular dystrophies. Recent and ongoing investigations, often cued by tumor biology, have revealed an important role for various growth factors, particularly in the neovascular form of the condition. Transgenic and knockout studies have provided important mechanistic insights into the development of choroidal neovascularization, the principal cause of vision loss in age-related macular degeneration. This in turn has culminated in preclinical and clinical trials of directed molecular interventions. PMID- 12745004 TI - Primary open-angle glaucoma in blacks: a review. AB - Glaucoma is one of the leading causes of blindness worldwide. Primary open-angle glaucoma (POAG) is the most prevalent form of glaucoma and has a particularly devastating impact in blacks. In the black American population, POAG prevalence is estimated to be six times as high in certain age groups compared to whites. POAG is more likely to result in irreversible blindness, appears approximately 10 years earlier and progresses more rapidly in blacks than in whites. Racial differences in optic disk parameters have been reported and show that blacks have larger optic disks than whites. This finding is robust and may account for the reported differences in other optic disk parameters. The existence of racial differences in intraocular pressure remains to be demonstrated, as conflicting findings are reported in the literature. Intraocular pressure may actually be underestimated in blacks, perhaps because they have thinner corneas. The prevalence of diabetes and hypertension is higher in blacks than in whites, and although no causal relationship has been established between POAG and each of these systemic diseases, some reports suggest that they often occur together, perhaps through an indirect relationship with intraocular pressure. Compounding the problem, there is evidence that blacks are less responsive to both drug and surgical treatment for POAG. Finally, they often have reduced accessibility to treatment and are less aware of the risks of having POAG. This article provides a comprehensive review of the current knowledge pertaining to POAG in blacks. PMID- 12745005 TI - Antifibrotics and wound healing in glaucoma surgery. AB - When medical and laser therapy fail to control intraocular pressure, glaucoma filtration surgery needs to be performed. Glaucoma surgery is unique in that its success is linked to interruption of the wound-healing response in order to maintain patency of the new filtration pathway. In this article we will review the wound-healing pathway and the pharmacologic interventions that have been employed clinically and experimentally to interrupt wound healing, particularly steroids and the antifibrotic agents 5-fluorouracil and mitomycin C. A review of the published literature looking at use of these agents to enhance success as well as the associated complications are presented, critiqued, and interpreted in order to put the studies in proper perspective. Future directions and recommendations regarding use of these agents are available and an introduction to newer wound modulating agents such as anti-transforming growth factor beta 2 is presented. PMID- 12745006 TI - Double trouble. AB - A 78-year-old woman presented with vertical double vision. The Bielschowsky Parks' three-step test suggested a left inferior rectus muscle paresis. Diagnostic testing confirmed the diagnosis of myasthenia gravis. Despite treatment with pyridostigmine bromide (mestinon) the diplopia persisted. She refused corticosteroid treatment and sought a second opinion, resulting in three unsuccessful strabismus surgeries to correct her ocular misalignment. The evaluation and management of ocular myasthenia gravis is reviewed and the appropriate timing of strabismus surgery is discussed. PMID- 12745007 TI - Escher and the ophthalmologist. AB - The Dutch graphic artist, Maurits C. Escher (1898-1972) is famous for intricate and sometimes illusory images which challenge our sensibility. Over many years, from the 1920s to the 1960s, he made designs with interlocking figures that confuse the distinction between object and background. His correspondence and writings suggest that these designs were largely self-created until the 1950s when fame brought him increasingly into contact with scholars from disciplines such as mathematics, crystallography, and psychology. One of these contacts was with an ophthalmologist, Johan W. Wagenaar (1911-), who had been using Escher's designs to illustrate lectures about vision during night driving. A correspondence began that extended for almost a decade and altered Escher's concept of his own work. It is an intriguing footnote to the career of this extraordinary artist. PMID- 12745008 TI - Maloccurrence in oculoplastic surgery related to the managed care environment. AB - Conflicts between specialties abound in problems of the periorbita with different specialties claiming expertise in this area. These conflicts include both the diagnosis and the treatment of oculoplastic problems. Cases are presented in which such conflicts have arisen leading to less than optimal care for the patients. Engendering these conflicts are the Managed Care fiats of both insurance companies and hospital territorial conflicts. Ideas are presented for the ethical resolution of both the physician conflicts and for the recognition that managed care induced conflicts lead ultimately to less than optimal care. PMID- 12745012 TI - Synthesis of 2'-deoxy-2'-[18F]fluoro-beta-D-arabinofuranosyl nucleosides, [18F]FAU, [18F]FMAU, [18F]FBAU and [18F]FIAU, as potential PET agents for imaging cellular proliferation. Synthesis of [18F]labelled FAU, FMAU, FBAU, FIAU. AB - An efficient and reliable synthesis of 2'-deoxy-2'-[(18)F]fluoro-beta-D arabinofuranosyl nucleosides is presented. Overall decay-corrected radiochemical yields of 35-45% of 4 analogs, FAU, FMAU, FBAU and FIAU are routinely obtained in >98% radiochemical purity and with specific activities of greater than 3 Ci/micromol (110 MBq/micromol) in a synthesis time of approximately 3 hours. When approximately 220 mCi (8.15 GBq) of starting [(18)F]fluoride is used, 25 -30 mCi (0.93 -1.11 GBq) of product (enough to image two patients sequentially) is typically obtained. PMID- 12745013 TI - Biodistribution of [111In-DTPA-D-Phe1]-octreotide in dogs: uptake in the stomach and intestines but not in the spleen points towards interspecies differences. AB - The purpose of the present study was to establish the tissue distribution in abdominal organs and the excretion of radioactivity after intravenous administration of [(111)In-DTPA-D-Phe(1)]-octreotide in healthy dogs. In five Beagle dogs computed tomography and single photon emission computed tomography (SPECT) at 24 h after injection of [(111)In-DTPA-D-Phe(1)]-octreotide revealed accumulation of radioactivity in the kidneys, gall bladder, gastric fundus and cardia, intestinal tract, but not in the spleen. These findings were confirmed by in vitro scintigraphy of single abdominal organs. This also demonstrated accumulation of radioactivity in the pancreas and located the radioactivity in the gastrointestinal tract primarily in the wall itself. In vitro autoradiography with (125)I-[Tyr(3)]-octreotide on tissue samples in two dogs revealed sst receptors in the medullary part of the kidney, the basal two-thirds of the gastric mucosa of the cardia and fundus, Peyer's patches and neural plexus of the gastrointestinal tract. No sst receptors were demonstrated in the liver, spleen, and pancreas. These results differ to findings in man, where there is uptake in the spleen but not in the stomach, most likely caused by interspecies variations in sst receptor subtype expression. PMID- 12745014 TI - Determination of [18F]FCWAY, [18F]FP-TZTP, and their metabolites in plasma using rapid and efficient liquid-liquid and solid phase extractions. AB - Liquid-liquid and solid phase extraction methods were developed for the accurate and rapid quantitation of radioactive components in human plasma following injection of two PET ligands. A solid phase extraction (SPE) method was developed for the determination of the 5HT(1A) receptor ligand [N-[2-[4-(2-methoxyphenyl) piperazino]ethyl]-N-(2-pyridinyl) trans-4-[(18)F]fluorocyclohexanecarboxamide (FCWAY), and its acidic metabolite, 4-[(18)F]fluorocyclohexane carboxylic acid (FC). In both cases, the extraction method was much faster and easier to use, yet provided results comparable to HPLC and TLC methods. In addition, an easy to perform two-step liquid-liquid extraction was developed for quantitation of 3-(3 ((3-[(18)F]fluoropropyl)thio)-1,2,5-thiadiazol-4-yl)-1,2,5,6-tetrahydro-1 methylpyridine ([(18)F]FP-TZTP), a selective M2 muscarinic agonist. PMID- 12745015 TI - Aminocarboxylate complexes and octreotide complexes with no carrier added 177Lu, 166Ho and 149Pm. AB - Several aminocarboxylate complexes of the "no carrier added" (NCA) radiolanthanides (149)Pm, (166)Ho and (177)Lu were evaluated using our in vitro hydroxyapatite and serum stability model and in vivo in normal CF-1 mice [10]. The aminocarboxylate chelates evaluated with the NCA radiolanthanides for in vitro stability were EDTA, CDTA, DTPA, MA-DTPA and DOTA. In addition, the NCA radiolanthanide complexes with DTPA-octreotide (DTPA-OCT) were synthesized and evaluated, as a model for a peptide conjugated aminocarboxylate complex. The biodistribution studies of the NCA complexes with DTPA, DOTA and DTPA-OCT showed that the in vitro model correctly predicted the in vivo stability of the radiolanthanide complexes, with Ln-DOTA > Ln-DTPA > Ln-DTPA-OCT. PMID- 12745016 TI - Biodistribution of 111in-DTPA-D-Phe1-octreotide in tumor-bearing nude mice: influence of amount injected and route of administration. AB - In nude mice carrying the human carcionoid GOT1 different amounts of (111)In-DTPA Phe(1)-octreotide and routes of administration were studied in relation to uptake in tumor and normal organs. The relative organ uptake varied with given amount; highest in tumor after 0.1 and 1 microg and lowest in muscle, heart and blood after 0.1 microg. The uptake decreased in lungs and spleen with higher amounts of (111)In-DTPA-Phe(1)-octreotide. In all organs studied the tumor-to-normal-tissue activity concentration ratio was maximal at 0.1 and 1 microg, but route of administration influenced the uptake only little. PMID- 12745017 TI - Receptor mediated uptake of a radiolabeled contrast agent sensitive to beta galactosidase activity. AB - Radiolabeling of the MRI contrast agent 1-[2-(beta-galactopyranosyloxy)propyl] 4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane with (111)In, and its evaluation is reported. Radiolabeling was performed in acetate buffer with 50-78% radiochemical yield. In vitro studies revealed that the asialoglycoprotein receptor-poor cell line MH1C1 has low uptake, while the receptor-rich cell lines BNL-CL2 and Hep G2 have higher uptake. In vivo, the uptake of the compound in receptor-rich organ liver was very high. Blocking the receptor in vivo, reduced liver uptake by 90% suggesting that the compound localizes in receptor-enriched tissues by binding to the asialoglycoprotein receptor. PMID- 12745018 TI - Tc-99m-labeled C5a and C5a des Arg74 for infection imaging. AB - The complement anaphylatoxin C5a and its natural metabolite C5a des Arg(74) (C5adR) are involved in several stages of the inflammatory process. Both act on a common receptor expressed on different cell types, including neutrophils and monocytes. The receptor binding affinity of C5a is in the nanomolar range and exceeds that of C5adR by 1-2 orders of magnitude. The biologic potency of C5a is considerably higher than that of C5adR. Here we tested both proteins labeled with (99m)Tc for imaging of infection. METHODS: The proteins were labeled with (99m)Tc via the hydrazinonicotinamide (HYNIC) chelator. The preparations were tested for imaging of infection in a rabbit model of intramuscular infection. Biodistribution of the radiolabel was determined by gamma-camera imaging and by counting dissected tissues at 5 h p.i. RESULTS: C5a and C5adR showed in vivo abscess uptakes of 0.12 and 0.025%ID/g, abscess/muscle ratios of 76 and 14, abscess/blood ratios of 9.1 and 2.6, and ROI derived target-to-background ratios of 5.9 and 2.1, respectively at 5 h p.i. CONCLUSION: For infection imaging (99m)Tc-labeled C5a showed excellent in vivo characteristics. However, C5a is a very bioactive protein, impeding its clinical use as an infection imaging agent. The naturally occurring partial agonist C5adR has less biological effect but showed suboptimal imaging characteristics. The present study showed that for adequate localization of a receptor binding ligand affinities for the receptor in the nanomolar range are required. PMID- 12745019 TI - Synthesis and evaluation of 11C- and 18F-labeled 1-[2-(4-alkoxy-3 methoxyphenyl)ethyl]-4-(3-phenylpropyl)piperazines as sigma receptor ligands for positron emission tomography studies. AB - We prepared sigma(1)-receptor selective 1-([4-methoxy-(11)C]-3,4 dimethoxyphenethyl)-4-(3-phenylpropyl)piperazine ([(11)C]SA4503) and its fluorinated analog 1-([4-methoxy-(11)C]3,4-dimethoxyphenethyl)-4-[3-(4 fluorophenyl)propyl]piperazine ([(11)C]SA5845), and their [(11)C]ethoxy and [(18)F]fluoroethoxy analogs, and evaluated their potential for positron emission tomography studies. [(11)C]SA4503 is most selective for sigma(1) receptors, and the other five showed affinities for sigma(1) and sigma(2) receptors with a different extent. All radioligands showed the receptor-specific binding in the brain, and visualized similar regional brain distributions by ex vivo autoradiography. The [(11)C]ethoxy analogs were relatively labile for metabolism. PMID- 12745020 TI - Synthesis of fluorine-18-labeled ciprofloxacin for PET studies in humans. AB - Ciprofloxacin (1-cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-7-(1-piperazinyl) quinoline-3-carboxylic acid), a widely-prescribed antibiotic, was labeled with fluorine-18 with the aim to perform positron emission tomography studies in humans for pharmacokinetic measurements. Due to a lack of chemical activation of ciprofloxacin for a direct nucleophilic exchange reaction a novel two-step synthetic approach, which employed an activated 6-fluoro-7-chloro substituted precursor molecule, was developed. The radiosynthesis yielded, starting from 52.5 +/- 11.3 GBq of [(18)F]fluoride, 1.3 +/- 0.6 GBq (n = 13) [(18)F]ciprofloxacin ready for intravenous administration in about 130 min synthesis time. A series of analytical tests was performed in order to prove the identity of the radiolabeled compound and its suitability for human applications. PMID- 12745021 TI - N-methylpiperidinemethyl, N-methylpyrrolidyl and N-methylpyrrolidinemethyl esters as PET radiotracers for acetylcholinesterase activity. AB - The N-[(11)C]methylpiperidinyl esters are used as radiopharmaceuticals for measuring brain cholinesterase activity. We have synthesized a series of N methylpiperidinemethyl (1), N-methylpyrrolidinyl (2) and N methylpyrrolidinemethyl (3) esters and examined the effects of sterric constraint and stereochemistry on cholinesterase-mediated cleavage. Acetylcholinesterase exhibited a preference for primary esters 1 and for the R-isomers of both 1 and 2. Biological data for (S)-N-[(11)C]methyl-2-piperidinemethyl acetate (1a) were similar to [(11)C]AMP. These data better define the structure-activity relationships for cholinesterase radiotracers and provide lead compounds for (18)F- labeling. PMID- 12745022 TI - Cellular retention of radioactivity and increased radiation dose. Model experiments with EGF-dextran. AB - Targeting of tumor cells with radiolabeled biomolecules is a possible approach to inactivate disseminated tumor cells. However, rapid degradation of the biomolecules after cellular internalization and subsequent excretion of the radioactivity is a problem. We studied the possibility of using dextran as a carrier of radionuclides to improve the intracellular retention. An EGF-dextran conjugate, aimed for targeting of tumor cells overexpressing the EGF-receptor, was used as model. Retention tests were performed with (125)I on different parts: [(125)I]-EGF-dextran-[(125)I], [(125)I]-EGF-dextran and EGF-dextran-[(125)I]. Comparisons were made with [(125)I]-EGF. The radiolabeled compounds were incubated with cultured glioma cells for different times. The cellular retention of radioactivity was then measured for up to 24 h. Expected radiation doses at the cellular level were calculated assuming that (131)I, instead of (125)I, was coupled to EGF and EGF-dextran. The results indicated that the EGF-part of the conjugate was degraded and the EGF-attached radioactivity was rapidly excreted, whereas radioactivity on dextran was retained intracellularly to a high degree, i.e. 70-80% of the radioactivity bound to dextran was still cell-associated after 24 h. The retention after 24 h was significantly higher (p < 0.001) when the radioactivity was on the dextran instead of the EGF-part. The radiolabeled EGF dextran had a notably high specific radioactivity; up to 11 MBq/microg. There was potential for at least hundred times increased radiation dose per receptor interaction when the radioactivity was on the dextran part. The advantage with radioactivity on the dextran part was the high cellular retention and the high specific radioactivity (higher than previously reported for other residualizing labels) without severe loss of receptor specific binding. Thus, dextran seems suitable as a carrier of radionuclides aimed for therapy and gives potential for a highly increased radiation dose. PMID- 12745023 TI - [18F]Fluoroazomycinarabinofuranoside (18FAZA) and [18F]Fluoromisonidazole (18FMISO): a comparative study of their selective uptake in hypoxic cells and PET imaging in experimental rat tumors. AB - The present study compares the uptake of [(18)F]Fluoroazomycinarabinofuranoside ((18)FAZA), a recently developed hypoxia tracer for PET imaging of tissue hypoxia, with an established tracer [(18)F]Fluoromisonidazole ((18)FMISO) both in vitro, using Walker 256 rat carcinosarcoma cells, and in vivo in experimental rat tumors eleven to twelve days after tumor cell implantation. In vitro studies indicated that hypoxia-selective uptake of both (18)FAZA and (18)FMISO in tumor cells, 20 and 100 minutes post-incubation was of the same magnitude (20 min: 1.24 +/- 0.4% ((18)FAZA); 1.19 +/- 0.7% ((18)FMISO); 100 min: 3.6 +/- 1.6% ((18)FAZA); 3.3 +/- 1.7% ((18)FMISO)). PET imaging reflected a similar radiotracer distribution in rat tumors for (18)FAZA and (18)FMISO one h after radiotracer injection. The concentration of (18)FAZA in the tumors as measured by PET, however, was lower in comparison to (18)FMISO (SUV(FAZA) = 0.61 +/- 0.2 vs. SUV(FMISO) = 0.92 +/- 0.3, p < 0.05) although the tumor to muscle ratios for (18)FAZA and (18)FMISO did not differ in the PET images that were obtained after one h (SUV(FAZA) = 2.5 +/- 0.5 vs. SUV(FMISO) = 2.9 +/- 0.7). A comparison of PET data three h post-injection (SUV(FAZA) = 3.0 +/- 0.5 vs. SUV(FMISO) = 4.6 +/- 1.8, p < 0.05) demonstrated a lower (18)FAZA uptake that indicates a lower sensitivity of (18)FAZA in comparison to (18)FMISO in detecting hypoxic regions at a longer time in this animal model. However, these data also show a faster elimination of (18)FAZA from blood, viscera and muscle tissue, via the renal system. This advantage of a faster reduction of unspecific binding, in light of similar or marginally lower tumor uptake, warrants further investigation of (18)FAZA as a marker of regional hypoxia in tumors. PMID- 12745024 TI - In vivo recognition of cyclopentadienyltricarbonylrhenium (CpTR) derivatives. AB - In vivo metabolism of [(188)Re]tricarbonyl(carboxycyclopentadienyl)rhenium ([(188)Re]CpTR-COOH) and its glycine conjugate ([(188)Re]CpTR-Gly) was investigated to estimate the applicability of cyclopentadienyltricarbonylrhenium (CpTR) compounds to (186/188)Re-labeling reagents for polypeptides and peptides. Both [(188)Re]CpTR derivatives were stable after incubation in a buffered solution and in murine plasma at 37 degrees C for 6 h. Plasma protein binding was hardly observed with the two derivatives. However, different biodistribution and metabolic fates were observed with the two CpTR derivatives. While more lipophilic [(188)Re]CpTR-COOH was excreted by both hepatobiliary and urinary excretion, the majority of less lipophilic [(188)Re]CpTR-Gly was excreted by urinary excretion. In addition, while [(188)Re]CpTR-Gly was rapidly excreted into urine as its intact structure, [(188)Re]CpTR-COOH was metabolized to more hydrophilic compounds including its glycine conjugate, [(188)Re]CpTR-Gly. Renal excretion of [(188)Re]CpTR-Gly was significantly reduced in probenecid retreated mice. The present studies reinforced that CpTR core remained stable under biological environment. CpTR-COOH was partially recognized as an aromatic acid and was metabolized as such. However, glycine conjugation rendered CpTR-COOH hydrophilic enough to be excreted into urine without further metabolism. These findings suggested that radiolabeling reagents that liberate [(186/188)Re]CpTR Gly from covalently conjugated (186/188)Re-labeled polypeptides and peptides by the action of renal brush border enzymes would be useful to reduce renal radioactivity levels. PMID- 12745025 TI - Automated synthesis of the ultra high specific activity of [11C]Ro15-4513 and its application in an extremely low concentration region to an ARG study. AB - We have designed and constructed an automated device for the production of ultra high specific activity (11)C-labeled compounds via [(11)C]CH(3)I synthesized by the single pass I(2) method. The optimum condition for the production of [(11)C]CH(3)I was determined to be 630 degrees C for oven-1 (reaction column), 50 degrees C for oven-2 (iodine column) and 50 ml/min for the He gas flow rate, and gave the maximum conversion ratio of [(11)C]CH(3)I, 44%. [(11)C]Ro15-4513, known as an inverse agonist of the benzodiazepine receptor, was produced under optimized conditions. An i.v. injectable [(11)C]Ro15-4513 solution of 1500 +/- 490 MBq (n = 6) with specific activity 4700 +/- 2500 GBq/micromol and a radiochemical purity of 98.2 +/- 2% was obtained automatically within 25 minutes (from EOB) by irradiating nitrogen gas containing 5% H(2) with 18 MeV protons (14.2 MeV on target) at 20 microA for 20 minutes. The highest specific activity of 9700 GBq/micromol (at EOS) could be achieved, although the radiochemical purity was 92.4%. By the use of the ultra-high specific activity [(11)C]Ro15 4513, the super high affinity binding sites in the rat brain hippocampus could be clearly visualized even at the extremely low concentration of 0.66 pM Ro15-4513 by in vitro autoradiography. PMID- 12745026 TI - Culturally competent treatment of African American clients among a national sample of outpatient substance abuse treatment units. AB - This study measures the level of cultural competency with respect to African American clients that exists among a national sample of outpatient substance abuse treatment (OSAT) units and determines the relationship of cultural competency to various characteristics of these units and their clients. The study utilizes cross-sectional data from the 1995 National Drug Abuse Treatment System Survey (NDATSS). The sample for NDATSS was randomly selected from a comprehensive list of OSAT programs compiled by the Institute for Social Research at the University of Michigan in 1994. Of the nationally representative, stratified sample of 699 units, 618 (88%) participated. Spearman correlation, analysis of variance, Behrens-Fisher t-tests, and chi-square were used for bivariate comparisons. Culturally competent units are typically public, federal-funded organizations. Staffs of culturally competent units are typically college educated with specialized treatment certification. High severity of illness as well as increased social distresses is pervasive among the clients of culturally competent units. Consideration of this client profile may be a key determinant in evaluating the effectiveness of cultural competency for African American substance abusers. PMID- 12745027 TI - Are the treatment goals of culturally competent outpatient substance abuse treatment units congruent with their client profile? AB - This study examines whether organizational treatment goals varied among outpatient substance abuse treatment (OSAT) units identified as providing a level of culturally competent care for African Americans. Cross-sectional data from the 1995 National Drug Abuse Treatment System Survey (NDATSS) was used. Of the nationally representative, stratified sample of 699 units, 618 (88%) participated. Spearman correlation, analysis of variance, Behrens-Fisher t-tests, chi-square, and multivariate logistic regression were used. Culturally competent units, even after controlling for organizational and client characteristics, were more apt than non-culturally competent units to indicate that achieving steady employment, spiritual strength, and physical health were important treatment goals. A congruency exists among culturally competent OSAT units between the client profile, which is more distressful than that for non-culturally competent units, and the orientation of treatment goals, which are more holistic; that is, treating the total individual, rather than the addiction only. PMID- 12745028 TI - Stability and change in dimensional ratings of personality disorders in drug abuse patients during treatment. AB - The objective of the study was to determine the extent of change in dimensional scores of personality disorders during treatment of drug abuse patients. The drug abuse patients were monitored prospectively during treatment. Over a period of 6 years, at 3-monthly intervals, all residents in a therapeutic community for drug abuse patients were administered the Millon Clinical Multiaxial Inventory-II (MCMI-II), a questionnaire developed to measure Diagnostic and Statistical Manual of Mental Disorders (3rd ed., revised) personality disorders. Subjects who completed the MCMI-II at least at four different times were selected for this study (n = 72). Results of the study showed that treatment had resulted in significant changes in the dimensional scores of some personality disorders, whereas other dimensional scores did not change at all. Implications for treatment are discussed. PMID- 12745029 TI - Deception in prison assessment of substance abuse. AB - The relationship of The Paulhus Deception Scales (PDS; ) to the Texas Christian University Drug Screen I was investigated in participants screened for a voluntary in-prison substance abuse treatment program. Mean PDS scores for 35 female and 277 male prison inmates were significantly higher than the inmate norms in the test manual. All three PDS scale scores were negatively related to symptom endorsement and to the diagnosis of drug dependence. Almost 37% of participants produced protocols of questionable validity, with 22% faking-good and 14.7% faking-bad. However, over 90% of participants obtained scores that were not covered by the manual's profile typology. These findings underline the importance of deception to correctional assessment and the need for more information on the psychometrics and operating characteristics of the PDS in correctional settings. PMID- 12745030 TI - Retaining counseling staff at substance abuse treatment centers: effects of management practices. AB - The turbulence created by the emergence of managed care has increased the importance of staff retention for substance abuse treatment centers. This study examines the relationships between management practices, organizational commitment, and turnover intention among substance abuse treatment counselors. Counselors from 345 randomly selected privately funded treatment centers were surveyed about management practices, organizational commitment, and turnover intention. Structural equation model estimates indicate significant associations between these variables. The effects of performance-based rewards and support for creativity, two measures of management practices, on turnover intention are mediated by organizational commitment. Job autonomy has a direct effect on turnover intention. Treatment center administrators may improve staff retention by systematic attention to these factors shown to enhance organizational commitment and reduce counselors' intentions to quit. Such steps may include increasing counselor autonomy, providing rewards for strong job performance, and establishing a work environment that supports creativity and innovation. PMID- 12745031 TI - Responsivity: the value of providing intensive services to high-risk offenders. AB - Tests of the importance of service matching have had varying results, yet little attention has been given to testing the hypotheses about outcomes for clients based on differing risks to recidivate. We set out to test the risk and responsivity principles using a sample of clients from one site of a four-site randomized block experimental design study examining the effectiveness of a seamless system approach vs. traditional criminal justice supervision. Findings from our preliminary examination of official and self-report data from this site suggest the importance of the risk and responsivity concepts in providing substance abuse treatment, particularly for high-risk clients. Because of the relatively low statistical power of the tests employed in this exploratory analysis, many observed relationships were not statistically significant. Nonetheless, findings offer important implications for the delivery of substance abuse treatment. Future analyses are also recommended to further explore the impact of different service delivery systems. PMID- 12745032 TI - Gender differences in outcomes from prison-based residential treatment. AB - This study examines gender similarities and differences in background characteristics, the effectiveness of treatment, and the predictors of post release outcomes among incarcerated drug-using offenders. The sample of 1,842 male and 473 female treatment and comparison subjects came from a multi-site evaluation of prison-based substance abuse treatment programs. Three-year follow up data for recidivism and post-release drug use were analyzed using survival analysis methods. Despite the greater number of life problems among women than men, women had lower three-year recidivism rates and rates of post-release drug use than did men. For both men and women, treated subjects had longer survival times than those who were not treated. There were both similarities and differences with respect to gender and the other predictors of the two post release outcomes. Differences in background characteristics and in factors related to post-release outcomes for men and women suggest the plausibility of gender-specific paths in the recovery process. PMID- 12745033 TI - Internet recovery for substance abuse and alcoholism: an exploratory study of service users. AB - The Internet provides accessible, available, and affordable services for a variety of personal health issues. Use of Internet-based support tools for personal recovery from substance abuse, alcoholism, and mental illness is not new; however, several factors appear to have stalled investigation of the use of these tools in the prevention, outreach, support, recovery, and aftercare for those affected by substance abuse and/or alcoholism. This study was undertaken to begin the process of identifying Internet recovery services (IRS) and describing users of these services. Basic demographic and program affiliation data were obtained via an Internet survey utilizing conservative methodology. An obtained sample of more than 1000 surveys yielded a usable data analysis sample of 928, indicating the widespread use of IRS and diversity of service users, which includes all ethnic groups and age levels within the United States, using more than 70 different recovery programs and services. Future research needs are discussed. PMID- 12745034 TI - Perceived barriers to quitting smoking among alcohol dependent patients in treatment. AB - Little is known about the perceived barriers to quitting smoking among alcohol abusers. In addition to the usual barriers perceived by smokers, alcohol dependent smokers may have a few barriers unique to their addictive lifestyle. The Barriers to Quitting Smoking in Substance Abuse Treatment (BQS-SAT) was administered to 96 alcohol dependent smokers in residential substance abuse treatment. The BQS-SAT is designed to assess perceived barriers to quitting smoking among alcohol abusers using eleven true-false items. One open-ended item was included to gather information about potential additional barriers. The majority of respondents reported withdrawal-related barriers such as expecting to feel irritable, anxious, restless, and about half expected intolerable urges to smoke if they were to quit smoking, as most smokers do. However, concerns about effects on sobriety and needing cigarettes to cope with feeling down were also endorsed by almost half of the patients. Total number of perceived barriers was significantly related to smoking history, expected effects from smoking, and smoking temptation but was not associated with severity of alcohol use or dependence on admission. Providing corrective feedback about these barriers could be useful when addressing smoking with patients who have alcohol abuse or dependence. PMID- 12745035 TI - Prospective evaluation of emergency department patients with potential coronary syndromes using initial absolute CK-MB vs. CK-MB relative index. AB - We compared the predictive properties of an initial absolute creatine kinase-MB (CK-MB) to creatine kinase-MB relative index (CK-MB RI) for detecting acute myocardial infarction (AMI), acute coronary syndromes (ACS), and serious cardiac events (SCE). Consecutive patients > 24 years of age with chest pain who received an electrocardiogram (EKG) as part of their Emergency Department (ED) evaluation had CK and CK-MB drawn at presentation. Patients were followed prospectively during their hospital course. The main outcome was AMI, ACS or SCE (death, AMI, dysrhythmias, CHF, PTCA/stent, CABG) within 30 days. The sensitivity, specificity, PPV and NPV of CK-MB and CK-MB RI to predict AMI, ACS, and SCE were calculated with 95% CIs. We enrolled 2028 patients. There were 105 patients (5.2%) with AMI, 266 (13.1%) with ACS, and 150 with SCE (7.4%). Absolute CK-MB had a higher sensitivity than CK-MB RI for AMI (52.0 vs. 46.9, respectively), ACS (23.5 vs. 20.8, respectively), and SCE (39.6 vs. 36.0, respectively), but a lower specificity than CK-MB RI for AMI (93.2 vs. 96.1, respectively), ACS (93.1 vs. 96.1, respectively) and SCE (93.3 vs. 96.3, respectively); and lower PPV for AMI (35.7 vs. 46.5, respectively), ACS (42.0 vs. 53.4, respectively) and SCE (38.5 vs. 50.5, respectively). The negative predictive values were similar for all outcomes. We conclude that the risk stratification of ED chest pain patients by absolute CK-MB has higher sensitivity, similar NPV, but a lower specificity and PPV than CK-MB relative index for detection of AMI, ACS, and SCE. The optimal test depends upon the relative importance of the sensitivity or specificity for clinical decision-making in an individual patient. PMID- 12745036 TI - Frequency of acute coronary syndrome in patients presenting to the emergency department with chest pain after methamphetamine use. AB - We reviewed the frequency of acute coronary syndrome (ACS) in patients presenting to our emergency department (ED) with chest pain after methamphetamine (MAP) use during a 2-year interval. Thirty-three patients (25 males, 8 females; average age 40.4 +/- 8.0 years) with a total of 36 visits met study inclusion criteria: 1) non-traumatic chest pain, 2) positive MAP urine toxicology screen, 3) admission to "rule-out" myocardial infarction, 4) chest radiograph demonstrating no infiltrates. An ACS was diagnosed in 9 patients (25%). Three patients (8%) (2 ACS and 1 non-ACS) suffered cardiac complications (ventricular fibrillation, ventricular tachycardia, supraventricular tachycardia, respectively). Age, gender, cardiac risk factors, prior coronary artery disease, initial systolic blood pressure and heart rate did not differ significantly in the ACS and non-ACS groups. The initial and subsequent electrocardiograms (EKG) were normal in 1/9 (11%) patients with ACS and 16/27 (59%) without ACS (p < 0.05). Our findings suggest that: 1) ACS is common in patients hospitalized for chest pain after MAP use, and 2) the frequency of other potentially life-threatening cardiac complications is not negligible. A normal EKG lowers the likelihood of ACS, but an abnormal EKG is not helpful in distinguishing patients with or without ACS. PMID- 12745037 TI - Goal-directed abdominal ultrasonography: impact on real-time decision making in the emergency department. AB - The impact of "goal-directed" abdominal ultrasound (US) on real-time decision making in the emergency department (ED) was studied, with specific emphasis on the certainty of diagnosis, treatment, and disposition plans. A prospective, interventional study enrolled 212 patients at a county teaching hospital ED, who underwent bedside US by experienced ED sonographers. A study questionnaire was completed documenting the US indication, working diagnosis, treatment, and disposition plan. The physicians assigned pre-test and post-test levels of certainty for the diagnosis, treatment plan, and disposition on an integral scale from 1 to 10. Scores for diagnosis were further categorized into low (1-3), moderate (4-7) and high certainty of disease. Absolute mean changes in level of certainty for diagnosis, treatment, and disposition were 3.2 (95% CI 3.1-3.3), 2.0 (95% CI 1.9-2.1), and 1.9 (95% CI 1.8-2.0), respectively. The direction of change after US for certainty of diagnosis was evenly split, with 47% increasing and 47% decreasing. The majority of patients categorized as either high or low certainty of disease had US results concordant with the physician's initial assessment. However, 16% moved from either high to low or from low to high certainty categories after US. Patients with moderate certainty moved evenly to either the low or high post-test category in 97% of cases. Treatment and disposition decisions were less impacted by US, with the majority of cases increasing in certainty irrespective of the US results. Bedside ultrasonography in the ED has an important impact on real-time decision-making, particularly in terms of the certainty of diagnosis. PMID- 12745038 TI - Beta-blocker use in the emergency department in patients with acute myocardial infarction undergoing primary angioplasty. AB - Our objectives were to evaluate the frequency of beta-blocker administration in the setting of acute myocardial infarction (AMI) where angioplasty is the primary treatment, and to investigate emergency physician's (EPs) attitudes toward beta blockers. We performed a retrospective chart review of all patients who presented with symptoms and electrocardiogram (EKG) criteria consistent with AMI in the defined study period. Charts were reviewed for beta-blocker administration and other treatments. A survey was subsequently distributed to all EPs to determine self-reported reasons for withholding beta-blockers. There were 91 patients identified. Of those who did not have contraindications, 99% (89/90) received aspirin, 97% (88/91) received heparin, 94% (84/89) received nitrates, but only 28% (19/68) received beta-blockers. Ninety-six percent of beta-blocker-eligible patients received them as inpatients. Eighty-six percent (44/52) of EPs completed the survey. Physicians felt strongly about avoiding beta-blockers in patients with asthma exacerbation, severe congestive heart failure, and high degree AV block. Bradycardia was the most frequent reason for withholding beta-blockers. In this series of patients presenting with AMI, beta-blockers were greatly underutilized. The self-reported reasons of EPs for withholding beta-blocker therapy did not explain why 72% (49/68) of patients without contraindications did not receive beta-blockers. PMID- 12745040 TI - Anticoagulation and spontaneous retropharyngeal hematoma. AB - A 57-year-old man treated with Coumadin for an episode of paroxysmal atrial fibrillation presented to the Emergency Department with progressive dysphagia and odynophagia. Physical examination revealed a large right-sided lateral and posterior pharyngeal wall mass. Laboratory studies were significant for an international normalized ratio (INR) of 5.4 with a white blood cell count of 11,600/muL and a hematocrit of 33.2%. A lateral soft tissue radiograph and computed tomography (CT) scan of the neck demonstrated a retropharyngeal hematoma compromising the airway. Rapid reversal of the coagulopathy was achieved with fresh frozen plasma and vitamin K. Patients with space-occupying retropharyngeal masses present a significant management dilemma. The choice between observation, intubation, or a surgical airway intervention must rely upon knowledge of the natural course and high rate of airway occlusion when faced with a patient who presents with a retropharyngeal hematoma. Once symptoms of airway compromise manifest, the airway must be expeditiously secured before complete obstruction occurs. This diagnosis should be sought when an anticoagulated patient presents with upper airway symptoms. PMID- 12745039 TI - High altitude headache: efficacy of acetaminophen vs. ibuprofen in a randomized, controlled trial. AB - Ibuprofen has been shown to be more effective than placebo in the treatment of high altitude headache (HAH), but nonsteroidal anti-inflammatory agents have been linked to increased incidence of gastrointestinal (GI) side effects and high altitude pulmonary edema (HAPE). We postulated that acetaminophen, which does not share ibuprofen's theorized causal link to GI side effects or HAPE, could provide effective HAH therapy. We conducted a prospective, randomized, double-blind, clinical trial of ibuprofen vs. acetaminophen in the Solu Khumbu, Nepal: Mt. Everest Base Camp, Pheriche, Dingboche (4240 m to 5315 m). Seventy-four consecutive patients (ages 13 to 61 years) were randomized, were assessed with the Lake Louise Acute Mountain Sickness (AMS) criteria, and received a physical examination (which included vital signs, oxygen saturation as measured by pulse oximetry (SpO(2)), and assessment of clinical Lake Louise AMS criteria). Patients then received either 400 mg of ibuprofen (IBU) or 1000 mg of acetaminophen (ACET), and were asked to rate their cephalgia using a 10-cm visual analog scale (VAS). Thirty-nine patients received IBU, and 35 received ACET. Baseline Lake Louise AMS scores were identical in the two groups (mean = 5.9). No differences in mean VAS scores between IBU and ACET groups were noted at time 0 (presentation), 30, 60, or 120 min. No cases of HAPE or high altitude cerebral edema were noted during the study period. In this study population, acetaminophen was as effective as ibuprofen in relieving the pain of HAH. PMID- 12745041 TI - Transient adult jejunal intussusception. AB - Intussusception is a rare cause of abdominal pain in adults. It occurs in fewer than 1% of all cases of adult small bowel obstruction. In the adult population, most cases are the result of some type of intestinal lesion like adhesions, melanomas, lipomas or adenomatous polyps. Idiopathic intussusceptions are an extremely rare occurrence in adults, comprising only 2-23% of diagnosed intussusceptions. This report describes two cases of transient, idiopathic adult jejunal intussusception in a 19-year-old woman and a 39-year-old man, both presenting to the same hospital 1 week apart. Both patients complained of nonspecific abdominal pain and nausea and were diagnosed with intussusception by computed tomography (CT) scan. In both cases, no underlying bowel abnormality was identified and neither required a bowel resection. This report discusses the common presentation, diagnosis, and treatment of adult intussusception and questions whether patients with transient intussusceptions require exploratory laparotomy. PMID- 12745042 TI - Atraumatic Clostridial myonecrosis: case report and literature review. AB - Clostridial myonecrosis (CM) is a rare, life-threatening infection that is most often associated with recent surgery or skeletal muscle trauma. It usually affects patients with some degree of underlying immunocompromise or vascular insufficiency. Occasionally, CM can occur at remote sites, with seeding from a gastrointestinal source in the setting of malignancy. We report a case of a 75 year-old man who developed rapidly progressive myonecrosis in the right shoulder, without prior trauma, caused by Clostridium septicum. On autopsy, this patient was found to have previously undiagnosed radiation colitis with ulcerations and abscess formation, secondary to recent prostate cancer radiation therapy. Although several case reports discuss CM in the setting of bowel malignancy, our case illustrates that non-malignant bowel inflammation may be a sufficient source for the infection. Clinical features of this uncommon disease are discussed, and the relevant literature is reviewed with regard to Clostridium septicum as an etiologic agent. PMID- 12745044 TI - Diagnosis and monitoring of hemorrhagic shock during the initial resuscitation of multiple trauma patients: a review. AB - The initial management of the multiple trauma victim requires evaluation for potential hemorrhage and ongoing monitoring to assess the efficacy of resuscitation and avoid complications related to hemorrhagic shock. A variety of strategies exist to assess circulatory status, including hemodynamic monitoring, tissue perfusion measurement, and use of serum markers of metabolism. We review available technologies used to assess fluid status and tissue perfusion in patients with blood loss or hemorrhagic shock, discuss how these methods can be used effectively and efficiently during initial trauma resuscitation to guide therapy and disposition, and suggest directions for future research to improve outcomes by providing more appropriate and timely care and avoiding unnecessary complications. PMID- 12745043 TI - Thrombotic thrombocytopenic purpura (TTP) presenting as pancreatitis. AB - Thrombotic thrombocytopenic purpura (TTP) is a rare clinical entity. It is a multi-systemic disorder characterized by a clinical pentad of thrombocytopenia, microangiopathic hemolytic anemia, diffuse and nonfocal neurologic symptoms, decreased renal function, and fever. Abdominal pain is an uncommon presenting symptom for TTP. Pancreatitis may occur from TTP or, in a few cases, may trigger TTP. The clinical diagnosis of TTP is generally difficult because there are many varied clinical presentations and the full expression of the pentad may be prolonged. However, once the diagnosis is suspected or confirmed, immediate plasmapherseis with plasma exchange must be performed to reduce the severe morbidity from neurologic disability. PMID- 12745045 TI - Intrathecal baclofen withdrawal mimicking sepsis. AB - Baclofen (Lioresal) is a drug of choice to treat spasticity and is increasingly being administered intrathecally via an implantable pump in cases refractory to oral therapy. Emergency physicians will likely treat patients with baclofen withdrawal or overdose as this treatment becomes more widespread. The syndrome of baclofen withdrawal presents with altered mental status, fever, tachycardia, hypertension or hypotension, seizures, and rebound spasticity, and may be fatal if not treated appropriately. Baclofen withdrawal may mimic other diseases including sepsis, meningitis, autonomic dysreflexia, malignant hyperthermia, or neuroleptic malignant syndrome. Treatment consists of supportive care, reinstitution of baclofen, benzodiazepines, and diagnosis and eventual repair of intrathecal pump and catheter malfunction. PMID- 12745046 TI - Tension pyopneumothorax in a child: a case report. AB - Pneumonia is an infection of the lung parenchyma that may result in pleural thickening, effusion, or an empyema. When there is air or gas in association with purulent exudate in the pleural cavity, a pyopneumothorax exists. The progression to pyopneumothorax under tension is extremely rare. We present a case of tension pyopneumothorax in a child. PMID- 12745047 TI - Methanol ingestion: prevention of toxic sequelae after massive ingestion. AB - Methanol ingestion, a rare but potentially fatal poisoning, is often difficult to diagnose in the emergency department (ED) and historically has been difficult to treat. In this article, we report a methanol ingestion with a blood concentration of 692 mg/dL, which was treated with 4-methylpyrazole (Fomepizole) and dialysis, without sequelae. To our knowledge, such a massive ingestion has never been treated with this modality without development of long-term disability. Another unusual feature of this case is the significantly elevated serum osmolal gap at presentation without elevation in anion gap, demonstrating the effects of co ingestion of ethanol. Additionally, there was a marked disparity between the patient's breath alcohol analyzer level and the blood ethanol concentration, illustrating the inability of the breath alcohol analyzer to differentiate between volatile alcohols. Treatment of the methanol-poisoned patient with Fomepizole is discussed. PMID- 12745048 TI - Comminuted fracture of the ulnar sesamoid of the metacarpophalangeal joint of the thumb: an uncommon injury. AB - Fracture of the ulnar sesamoid of the metacarpophalangeal joint of the thumb is rare. Injury may be caused as a result of hyperextension or, less frequently, direct trauma. A sesamoid may be incompletely ossified, making it difficult to diagnose the fracture on radiographs. Early recognition of this entity leads to appropriate management. The case of a 26-year-old man who sustained a hyperextension injury of the thumb is presented. Radiographs demonstrated a fracture of the ulnar sesamoid of the thumb metacarpophalangeal joint. The literature regarding this uncommon injury is discussed. PMID- 12745049 TI - Droperidol in the emergency department: is it safe? AB - Droperidol is an antipsychotic and antiemetic drug that has been used extensively by emergency physicians, psychiatrists, and anesthesiologists worldwide since 1967. It also has been used effectively for other diverse conditions, such as treatment of headache and vertigo. As of January 2001, Droperidol was no longer available in Europe after its founder, Janssen-Cilag Pharmaceuticals, discontinued its distribution. In December 2001, the United States Food and Drug Administration (FDA) placed a black box warning on the use of Droperidol in response to an association between Droperidol and fatal cardiac dysrhythmias, such as torsade de pointes, resulting from prolongation of the QT interval. In this review we closely examine the pharmacology, indications, use, and complications associated with Droperidol, and speculate on its future use in the Emergency Department. PMID- 12745050 TI - Blunt multi-system trauma. PMID- 12745051 TI - Delayed presentation of traumatic diaphragmatic hernia. PMID- 12745052 TI - Significance of appendicoliths in abdominal pain. PMID- 12745053 TI - The history of anthrax. AB - Anthrax, a potentially fatal infection, is a virulent and highly contagious disease. Descriptions of this disease begin in antiquity, with the best ancient account being by the Roman poet Virgil. During the 19th century, anthrax was the infection involved in several important medical developments. It served as the prototype for Koch's postulates regarding the causation of infectious disease. The first vaccine containing attenuated live organisms was Louis Pasteur's veterinary anthrax vaccine. In the 1900s, human inhalation anthrax occurred sporadically in the United States among textile and tanning workers, but the incidence of the illness had declined dramatically. An outbreak of inhalation anthrax occurred in Sverdlovsk near a Soviet military microbiology facility in 1979. This epidemic represented the largest documented outbreak of human inhalation anthrax in history. In October and November 2001, 22 cases of confirmed or suspected inhalation and cutaneous anthrax were reported associated with the intentional release of the organism in the United States. An additional case of cutaneous disease occurred in March of 2002. PMID- 12745054 TI - Anti-coagulation and spontaneous retropharyngeal hematoma. PMID- 12745055 TI - Very high serum aldesterone level in an infant who expired. PMID- 12745056 TI - Acute acetaminophen poisoning in children: kids aren't just little adults. PMID- 12745057 TI - Parapneumonic pleural effusion and empyema in childhood. PMID- 12745058 TI - Comment on Fox et al's presentation of two invasive group A streptococcal (GAS) infections. PMID- 12745059 TI - Comment on Cory J. Pitre's points about the need for student emergency medical societies. PMID- 12745060 TI - Signal transduction by receptor activator of nuclear factor kappa B in osteoclasts. AB - Receptor activator of nuclear factor kappa B (RANK) is a tumor necrosis factor receptor (TNFR) family protein that plays an important role in the regulation of bone and immune systems. Cellular responses to RANK ligand (RANKL) and the signal transduction pathways of RANK have been well characterized in osteoclasts and osteoclast precursor cells. RANKL induces the differentiation of osteoclast precursor cells and stimulates the resorption function and survival of mature osteoclasts. The RANK signaling mechanisms involved in these responses include the recruitment of TNF receptor-associated factor proteins, the activation of transcription factors (NF-kappaB, AP-1, and NFAT2), the cascades of mitogen activated protein kinases (ERK, JNK, and p38), and the induction of Src- and phosphatidylinositol 3-kinase-dependent Akt activation. Despite the identification of several molecular targets, a comprehensive understanding of RANK signaling requires further studies on more complicated issues such as the temporal and spatial pattern of the engagement of signaling molecules and the precise relationship between the signaling pathway and the cellular response. PMID- 12745061 TI - Antidyslipidemic action of fenofibrate in dyslipidemic-diabetic hamster model. AB - Fenofibrate is the ligand for PPARalpha subtype that mediates the action of its agonists' in lipid metabolism. How fibrate exerts hypolipidemic effect? The mechanism is studied in a newly developed high-fat fructose enriched diet induced dyslipidemia-diabetic hamster model. Fenofibrate lowered the basal plasma lipids like TC, TG, PL, FFA, glycerol, VLDL, and LDL, but HDL was increased. The activity of lipoprotein lipase in liver, adipose tissue, and small intestine was upregulated. However, that of triglyceride lipase was downregulated in liver. It has also improved the insulin secretion and plasma glucose lowering, caused by impairment in insulin secretion due to high-fat load. The drug was found effective in reducing body weight and diet due to rise in leptin level. Fenofibrate also enhanced the fecal excretion of total lipids, cholic acid, and deoxycholic acid probably by the activation of 7alpha cholesterol hydroxylase enzyme. Thus, causing broad-spectrum lipid lowering along with inhibition of hepatic lipid biosynthesis and maintaining lipid-glucose homeostasis. PMID- 12745062 TI - Functional diversity between Rho-kinase- and MLCK-mediated cytoskeletal actions in a myofibroblast-like hepatic stellate cell line. AB - Using a rat myofibroblast-like hepatic stellate cell line, we studied the actomyosin-based cytoskeletal actions mediated by Rho-kinase and/or myosin light chain kinase (MLCK). Calmodulin/MLCK inhibitors W-7 and ML-7 attenuated cell migration dose-relatedly at concentrations from 10(-6) to 10(-4)M and collagen gel-contraction by the cells at 10(-4)M, respectively. Rho-kinase inhibitors Y 27632 and HA1077 attenuated the gel-contraction at concentrations from 10(-6) to 10(-4) M, respectively. These Rho-kinase inhibitors attenuated cell migration at 10(-7)M but enhanced the migration at 10(-4)M, respectively. They altered cell morphology showing prominent peripheral actin bundles and sparse central stress fibers, in comparison with the calmodulin/MLCK inhibitors. Both ML-7 and Y-27632 attenuated phosphorylation of myosin regulatory light chain and cell attachment to extracellular substrate. ML-7 attenuated the activation of GTP-binding protein Rac, while Y-27632 did not. These findings suggest that the actomyosin-based cytoskeletal actions can be functionally diverse depending on the Rho-kinase mediated pathway and the MLCK-mediated pathway. PMID- 12745063 TI - Naringenin inhibits phosphoinositide 3-kinase activity and glucose uptake in 3T3 L1 adipocytes. AB - Previous studies have shown that flavonoids inhibit glucose uptake in cultured cells. In this report, we show that the grapefruit flavanone naringenin inhibited insulin-stimulated glucose uptake in 3T3-L1 adipocytes in a dose-dependent manner. Naringenin acts by inhibiting the activity of phosphoinositide 3-kinase (PI3K), a key regulator of insulin-induced GLUT4 translocation. Although naringenin did not alter the phosphotyrosine status of the insulin receptor, insulin receptor substrate proteins, or PI3K, it did inhibit the phosphorylation of the downstream signaling molecule Akt. In an in vitro kinase assay, naringenin inhibited PI3K activity. A physiologically attainable dose of 6 microM naringenin reduced insulin-stimulated glucose uptake by approximately 20%. This inhibitory effect remained 24h after the removal of naringenin from the culture medium. Collectively, our findings suggest that the regular consumption of naringenin in grapefruit may exacerbate insulin resistance in susceptible individuals via impaired glucose uptake in adipose tissue. PMID- 12745064 TI - Molecular characterisation of six alternatively spliced variants and a novel promoter in human peroxisome proliferator-activated receptor alpha. AB - Peroxisome proliferator-activated receptor alpha (PPARalpha) is a ligand activated transcriptional factor that governs many biological processes, including lipid metabolism, inflammation, and atherosclerosis. We demonstrate here the existence of six variants and multiple transcriptional start sites of the 5(') untranslated region (UTR) of hPPARalpha gene, originating from the use of alternative splicing mechanisms and four different promoters. Three new novel exons at the 5(')-untranslated region of human PPARalpha gene were also identified and designated as Exon A, Exon B, and Exon 2b. In addition, 1.2kb promoter fragment which drives the transcription of 2 variants with Exon B (hPPARalpha4 and 6) was successfully cloned and characterised. Sequencing results revealed promoter B did not contain a conservative TATA box within the first 100 nucleotides from transcriptional start site but has several GC-rich regions and putative Sp1 sites. Using luciferase reporter constructs transfected into HepG2 and Hep3B cell lines, promoter B was shown to be functionally active. Basal transcriptional activity was significantly high in the promoter fragment 341/+34, but lower in the region -341/-1147 as compared to the fragment -341/+34, indicating the presence of an element conferring transcriptional activation between positions -341 and +34 or alternatively, the presence of transcriptional repression between positions -341 and -1147 in the promoter B of hPPARalpha. PMID- 12745065 TI - Muscle type difference in the regulation of UCP3 under cold conditions. AB - We found opposite regulation of uncoupling protein 3 (UCP3) in slow-twitch soleus and fast-twitch gastrocnemius muscles of rats during cold exposure. Namely, the UCP3 mRNA level was downregulated in the soleus muscles, but upregulated in the gastrocnemius muscles after a 24-h cold exposure. In the analysis of UCP3 protein, we first succeeded in detecting UCP3 short-form as well as the long-form in vivo, which levels were decreased markedly in the cold-exposed soleus muscles. However, the levels of UCP3 and cytochrome oxidase subunit IV were well maintained in the cold-exposed gastrocnemius muscles with a rise in the total mitochondrial protein level, suggesting an increase of total oxidative ability. The fast-twitch muscle rather than the slow-twitch one may play an important role in adaptive responses, including thermogenesis under acute cold exposure. PMID- 12745066 TI - Tyrosinase protects human melanocytes from ROS-generating compounds. AB - The effects of two tetrahydroisoquinolines (TIQs), tetrahydropapaveroline (THP) and salsolinol (SAL), on human primary melanocytes were studied. These compounds are naturally occurring alkaloids deriving from the condensation of dopamine with aldehydes. The effects on the viability were studied by treating the cells with variable concentration of THP or SAL; both TIQs were well tolerated up to roughly 30 micro M. At higher concentrations, THP became overtly toxic while SAL showed no cytotoxic effect up to 100 micro M. TIQs treatment determined an impairment of intracellular activity of antioxidant enzymes, like SOD, DT-diaphorase, and glutathione peroxidase. A decrease of alpha-ketoglutarate dehydrogenase activity was also evidenced following TIQs treatment; a very strong diminution was found in THP-treated cells, whose viability was highly decreased. Both TIQs increased tyrosinase-specific mRNA transcription followed, in the case of SAL, by an activation of tyrosinase. In the presence of tyrosinase inhibitors TIQs treatment resulted in a sharp cytotoxic effect even at concentrations normally well tolerated. Taken together these data suggest that tyrosinase represents an outstanding protective mechanism against ROS-generating compounds, once primary detoxifying mechanisms are impaired or not available. PMID- 12745067 TI - Transcribed short tandem repeats occur in couples with strongly preferred registers. AB - Short tandem repeats (STRs) have been widely observed, but most STRs have no recognized organization or function. Here we show that for diverse mRNAs, 84% of (GC)(n) repeats were found unexpectedly coupled with another STR, (GU)(n). These STR couples exhibited preferred polarity and register. In 3(') untranslated mRNA sequences (UTRs) 100% of (GC)(n>6) repeats were tightly coupled with (GU)(n). For (GC)(n), stem folding energy correlated with the length and number of neighboring, non-folding (GU)(n) partners (p=0.014). Approximately 20% of (AU)(n>/=14) repeats were coupled with (GU)(n). The STR couple (AC)(n)(AG)(n) also exhibited polarity and register preferences. The sequence arrangement at STR couple joints was conserved rigorously, suggesting that these sequences were under selection pressure. Some STR couples may function as mRNA processing landmarks, based on alternative transcript comparisons. These observations suggest that some transcribed STRs may be functional UTR signals with predictable organization and usage patterns. PMID- 12745068 TI - Permeability of C2C12 myotube membranes is influenced by stretch velocity. AB - Mechanical signals are critical to the growth and maintenance of skeletal muscle, but the mechanism by which these signals are transduced by the cell remains unknown. This work examined the hypothesis that stretch conditions influence membrane permeability consistent with a role for membrane permeability in mechanotransduction. C2C12 myotubes were grown in conditions that encourage uniform alignment and subjected to uniform mechanical deformation in the presence of fluorescein labeled dextran to evaluate membrane permeability as a function of stretch amplitude and velocity. Within a physiologically relevant range of conditions, a complex interaction between the two aspects of stretch was observed, with velocity contributing most strongly at large stretch amplitudes. This suggests that membrane viscosity could contribute to mechanotransduction. PMID- 12745069 TI - Activation of Nrf2 and accumulation of ubiquitinated A170 by arsenic in osteoblasts. AB - Sub-lethal levels of arsenic induce upregulation of stress proteins. We here report for the first time that inorganic arsenic activates the transcription factor Nrf2, which controls the expression of oxidative stress-induced proteins. Treatment of cultured MC3T3-E1 osteoblasts with arsenite or arsenate induced increase of Nrf2, followed by transcriptional activation of target genes encoding HO-1, Prx I, and A170. We found that arsenate (200-800 micro M) only slightly increased the normal 60kDa A170 protein but markedly increased higher molecular mass forms of A170 (HMM-A170) that appeared as smeared bands. Arsenate also markedly increased ubiquitin-conjugated cellular proteins, suggesting that HMM A170 was one of the poly-ubiquitinated proteins. Arsenite (50-100 micro M) also induced accumulation of HMM-A170 and ubiquitin-conjugated proteins. These results provide the first direct evidence that toxic arsenics impair the normal function of A170. Our findings provide a potential diagnostic tool for monitoring biotoxicity in cells and tissues in response to arsenic compounds. PMID- 12745070 TI - Early embryonic lethality caused by targeted disruption of the mouse PHGPx gene. AB - Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is the only known intracellular antioxidant enzyme that can directly reduce lipid hydroperoxide in membrane. Mitochondrial and non-mitochondrial PHGPx and sperm nuclei GPx are transcribed from one gene by alternative transcription using different first exons Ia and Ib, respectively. To examine the role of PHGPx in development, we generated mice deficient in PHGPx by a targeted disruption of all exons of the PHGPx gene. Heterozygotes are viable, fertile, and appear normal, despite having decreased levels of three types of PHGPx mRNA and protein. Embryos homozygous for PHGPx-null die between 7.5 and 8.5 days post coitum (dpc), probably developing distal apoptosis. We examined the expression of PHGPx in mouse embryos using immunohistochemical analysis with anti-PHGPx mAb. The expression of PHGPx was detected in the embryonic ectoderm and the yolk sac membrane at 7.5dpc. The results demonstrated that PHGPx is expressed in early gastrulation stage at 7.5dpc and that the expression of PHGPx was essential for normal mouse development. PMID- 12745071 TI - Increase in IP3 and intracellular Ca2+ induced by phosphate depletion in LLC-PK 1 cells. AB - The mechanisms by which Pi depletion rapidly regulates gene expression and cellular function have not been clarified. Here, we found a rapid increase in intracellular ionized calcium [Ca(2+)](i) by phosphate depletion in LLC-PK(1) cells using confocal microscopy with the green-fluorescence protein based calcium indicator "yellow cameleon 2.1." The increase of [Ca(2+)](i) was observed in the presence or absence of extracellular Ca(2+). At the same time, an approximately twofold increase in intracellular inositol 1,4,5-triphosphate (IP(3)) occurred in response to the acute Pi depletion in the medium. Furthermore, 2 aminoethoxydiphenyl borate completely blocked the [Ca(2+)](i) increase induced by Pi depletion. These results suggest that Pi depletion causes IP(3)-mediated release of Ca(2+) from intracellular Ca(2+) pools and rapidly increases [Ca(2+)](i) in LLC-PK(1) cells. PMID- 12745072 TI - Increased methylation of endogenous 20-kDa protein in HIT beta-cell during insulin secretion. AB - Enzymatic methylation of endogenous proteins in clonal pancreatic beta-cell, HIT T15, was investigated. When cell extract incubated with S-adenosyl-L-[methyl 3H]methionine was subjected to SDS-PAGE followed by fluorography, endogenous 20 kDa protein was highly [methyl-3H]-labeled. The increase of methylation was correlated with insulin secretion, when the cells were treated with secretagogue; at 5.5mM glucose, insulin secretion increased by 2.5-fold, while the 20-kDa methylation to about 3.2-fold. In the case of forskolin, another secretagogue, at 0.1mM, the methylation increased by approximately 4.5-fold. This increase of 20 kDa methylation was inhibited when the cells were treated with 3mM EGTA to inhibit insulin secretion by depleting extracellular calcium ion, indicating intercausal relation between methylation and insulin secretion. The [methyl-3H] labeled amino acids were identified by thin layer chromatography as N(G) methylated arginines. While arginyl residues in Gly-Arg-Gly sequence are known to be posttranslationally methylated, a synthetic nonapeptide, GGRGRGRGG, competed with the 20-kDa methylation; at 1 and 10 micro M nonapeptides, 62% and 78% of 20 kDa methylation were inhibited, respectively. Furthermore, Western immunoblot analysis of HIT cell extract against GGRGRGRGG antibodies strongly immunoreacted with the 20-kDa protein. These results suggested that methylation of the endogenous 20-kDa protein might play some role in insulin secretion. PMID- 12745073 TI - Teleost fish spermatozoa contain a cytosolic protein factor that induces calcium release in sea urchin egg homogenates and triggers calcium oscillations when injected into mouse oocytes. AB - Established studies in a variety of organisms including amphibians, fish, ascidians, nemerteans, echinoderms, mammals, and even a species of flowering plant, clearly demonstrate that an increase in intracellular egg calcium is crucial to the process of egg activation at fertilization. In echinoderms, egg activation appears to involve an egg phospholipase C gamma (PLCgamma). However, numerous studies in mammalian species suggest that calcium is released from internal egg stores at fertilization by a sperm-derived cytosolic protein factor. Recent studies in the mouse have identified this sperm-derived factor as being a novel sperm-specific PLC isoform with distinctive properties (PLCzeta). Homologues of PLCzeta have since been isolated from human and cynomolgus monkey sperm. In addition, sperm factor activity has been detected in non-mammalian species such as chicken, Xenopus, and a flowering plant. Here we report evidence for the existence of a similar sperm-derived factor in a commercially important species of teleost fish, the Nile tilapia Oreochromis niloticus (L). Using an established bioassay for calcium release, the sea urchin egg homogenate, we demonstrate that protein extracts obtained from tilapia spermatozoa exhibit PLC activity similar to that seen in mammalian sperm extracts, and also induce calcium release when added directly to the homogenate. Further, tilapia sperm extracts induced calcium oscillations when injected into mouse oocytes. PMID- 12745074 TI - Fungicidal effect of indolicidin and its interaction with phospholipid membranes. AB - The fungicidal effect and mechanism of a tryptophan-rich 13-mer peptide, indolicidin derived from granules of bovine neutrophils, were investigated. Indolicidin displayed a strong fungicidal activity against various fungi. In order to understand the fungicidal mechanism(s) of indolicidin, we examined the interaction of indolicidin with the pathogenic fungus Trichosporon beigelii. Fluorescence confocal microscopy and flow cytometry analysis revealed that indolicidin acted rapidly on the plasma membrane of the fungal cells in an energy independent manner. This interaction is also dependent on the ionic environment. Furthermore, indolicidin caused significant morphological changes when tested for the membrane disrupting activity using liposomes (phosphatidylcholine/cholesterol; 10:1, w/w). The results suggest that indolicidin may exert its fungicidal activity by disrupting the structure of cell membranes, via direct interaction with the lipid bilayers, in a salt-dependent and energy-independent manner. PMID- 12745075 TI - Abolishment of the interaction between cyclin-dependent kinase 2 and Cdk associated protein phosphatase by a truncated KAP mutant. AB - The cyclin-dependent kinase (Cdk)-associated protein phosphatase (KAP) is a human dual-specificity protein phosphatase that dephosphorylates Cdk2 on a conserved threonine residue, T160, in a cyclin dependent manner. Several aberrant KAP transcripts with characteristic deletion regions have been identified in hepatocellular carcinoma tissues. In this report, we demonstrated that multiple aberrant KAP transcripts were also present in a hepatoblastoma cell line (HepG2), albeit harboring a totally different set of deletions. By performing yeast two hybrid and co-immunoprecipitation experiments, a KAP-Cdk2 interaction domain located in the amino acid 1-34 region was identified. This interaction domain was different from the major protein interface deduced from crystal structure analysis. Using a yeast three-hybrid system, it was shown that the presence of a truncated KAP mutant encoding this interaction domain abolished the wild-type KAP Cdk2 interaction. In conclusion, a previously unidentified KAP-Cdk2 interaction domain was discovered. Truncated KAP mutants containing this domain interfered with the wild-type KAP-Cdk2 interaction. PMID- 12745076 TI - Identification and characterization of the Cdc42-binding site of IQGAP1. AB - IQGAP1 is a multi-domained protein that integrates signaling of the Rho family GTPase Cdc42 with regulation of the cytoskeleton. Using SPOT analysis and in vitro peptide competition assays we have identified a 24 amino acid region of IQGAP1 that is necessary for Cdc42 binding. Both in vitro and in vivo analyses reveal that deletion of this sequence abolishes binding of IQGAP1 to Cdc42. In addition, the ability of IQGAP1 to increase the amount of active Cdc42 in cells is abrogated upon removal of this region. An IQGAP1 mutant lacking the Cdc42 binding site mislocalizes to the cell periphery. These observations specifically define a short sequence of IQGAP1 that is required for its interaction with Cdc42 and demonstrate that Cdc42 binding is necessary for the normal subcellular distribution of IQGAP1. PMID- 12745077 TI - Comprehensive mutagenesis of HIV-1 protease: a computational geometry approach. AB - A computational geometry technique based on Delaunay tessellation of protein structure, represented by C(alpha) atoms, is used to study effects of single residue mutations on sequence-structure compatibility in HIV-1 protease. Profiles of residue scores derived from the four-body statistical potential are constructed for all 1881 mutants of the HIV-1 protease monomer and compared with the profile of the wild-type protein. The profiles for an isolated monomer of HIV 1 protease and the identical monomer in a dimeric state with an inhibitor are analyzed to elucidate changes to structural stability. Protease residues shown to undergo the greatest impact are those forming the dimer interface and flap region, as well as those known to be involved in inhibitor binding. PMID- 12745078 TI - Genomic organization and characterization of the mouse ELYS gene. AB - Differentiation of hematopoietic stem cells into blood cells is controlled by several transcription factors. Recently, we identified a putative transcription factor, ELYS (for embryonic large molecule derived from yolk sac), using a subtraction strategy. During mouse embryogenesis, ELYS transcripts were predominantly expressed in hematopoietic tissues, such as the yolk sac, aorta gonad-mesonephros (AGM), and liver. Here, we report the cloning and characterization of the mouse ELYS gene. The ELYS gene spanned approximately 60kb encoding 36 exons, and was assigned between D1Mit315 and D1Mit458 markers in chromosome 1. The transcription initiation site was identified as the G residue located 670bp upstream of the translation start codon. A region downstream of the transcriptional start site contributed to high promoter activity. This region contained potential DNA elements for transcription factors such as GATA-1, -2, 3, heat shock factor (HSF) 2, and NF-kappaB, which are known to play important roles in hematopoietic events. PMID- 12745079 TI - Increase of human MTH1 and decrease of 8-hydroxydeoxyguanosine in leukocyte DNA by acute and chronic exercise in healthy male subjects. AB - To investigate the effects of exercise on oxidative DNA damage, we measured the levels of 8-hydroxydeoxyguanosine (8-OH-dG, 7,8,dihydro-8-oxo-deoxyguanosine), and mRNA of its sanitization enzyme, human MutT homologue (hMTH1), after mild acute exercise in healthy male subjects. The basal 8-OH-dG levels of physically active subjects were significantly lower than those of sedentary subjects. After mild exercise for 30min, the 8-OH-dG levels of the sedentary subjects significantly decreased. In correspondence with the change of 8-OH-dG levels, the hMTH1 mRNA levels of the physically active subjects were significantly higher than those of the sedentary subjects. The levels of hMTH1 mRNA of sedentary subjects significantly increased after mild exercise. Furthermore, there was a significant negative correlation between the levels of 8-OH-dG and hMTH1 mRNA. These results suggest that a mild exercise has a beneficial effect on the maintenance of low levels of 8-OH-dG probably by keeping the sanitization system at higher levels. PMID- 12745080 TI - Deactivation of ROCK-II by Y-27632 enhances basolateral pancreatic enzyme secretion and acute pancreatitis induced by CCK analogues. AB - In isolated rat pancreatic acini, protein expression of RhoA and Rho-associated kinase, ROCK-II, and the formation of immunocomplex of RhoA with ROCK-II were enhanced by CCK-8, carbachol, and the phorbol ester TPA. The ROCK-specific inhibitor, Y-27632, did not alter basal amylase secretion, whereas it potentiated CCK-stimulated pancreatic enzyme secretion in vitro. During caerulein-induced pancreatitis occurring in mice in vivo, Y-27632 enhanced serum amylase levels and the formation of interstitial edema and vacuolization at 12-18h after the first injection of caerulein. Y-27632 in turn inhibited the recovery of protein expression of ROCK-II at 18h after the first caerulein injection. These results suggest that RhoA and ROCK-II assemble normal CCK-stimulated pancreatic enzyme secretion and prevent caerulein-induced acute pancreatitis. PMID- 12745081 TI - Interaction of the deafness-dystonia protein DDP/TIMM8a with the signal transduction adaptor molecule STAM1. AB - The Mohr-Tranebjaerg-Jensen deafness-dystonia-optic atrophy protein DDP/TIMM8a is translated on cytoplasmic ribosomes but targeted ultimately to the mitochondrial intermembrane space, where it is involved in mitochondrial protein import. STAM1 is a cytoplasmic signal-transducing adaptor molecule implicated in cytokine signaling. We report here a direct interaction between DDP and STAM1, identified by yeast two-hybrid screening and confirmed by co-immunoprecipitation, fusion protein "pull downs," and nuclear redistribution assays. DDP coordinates Zn(2+), and Zn(2+) was found to stimulate the DDP-STAM1 interaction in vitro. Endogenous STAM1 localizes predominantly to early endosomes, and we found no evidence that STAM1 is imported into mitochondria in vitro. Thus, the DDP-STAM1 interaction likely occurs in the cytoplasm or at the mitochondrial outer membrane. The DDP STAM1 interaction requires a coiled-coil region in STAM1 that overlaps with the immunoreceptor tyrosine-based activation motif (ITAM), a region previously shown to be important for interaction with Jak2/3 and hepatocyte growth factor regulated tyrosine kinase substrate (Hrs). Thus, DDP binding may alter the interactions of STAM1 with several cytoplasmic proteins involved in cell signaling and endosomal trafficking. PMID- 12745082 TI - A novel cytomedical vehicle capable of protecting cells against complement. AB - We have developed "Cytomedicine," which consists of functional cells entrapped in semipermeable polymer, and previously reported that APA microcapsules could protect the entrapped cells from injury by cellular immune system. However, microencapsulated cells were not protected from humoral immune system. Here, we developed a novel APA microcapsule, in which APA microbeads (APA(Ba) microbeads) were modified to contain a barium alginate hydrogel within their centers in an attempt to make it more difficult for antibody and complement to permeate the microcapsules. The permeability of APA(Ba) microbeads was clearly less than that of APA microcapsules, presumably due to the presence of barium alginate hydrogel. Cells encapsulated within APA(Ba) microbeads were protected against treatment with xenogeneic anti-serum. Furthermore, murine pancreatic beta-cells encapsulated in APA(Ba) microbeads remained viable and continued to secrete insulin in response to glucose. Therefore, APA(Ba) microbeads may be a useful carrier for developing anti-complement device for cytomedical therapy. PMID- 12745083 TI - The viral death protein Apoptin interacts with Hippi, the protein interactor of Huntingtin-interacting protein 1. AB - Apoptin, a chicken anemia virus-encoded protein, induces apoptosis in human tumor cells but not in normal cells. The tumor-specific activity of Apoptin is correlated with its nuclear localization in tumor cells. In an attempt to elucidate the molecular mechanism of Apoptin-induced apoptosis, we identified human Hippi, the protein interactor and apoptosis co-mediator of Huntingtin interacting protein 1, as one of the Apoptin-associated proteins by yeast two hybrid screen. We also demonstrated that Hippi could interact with Apoptin both in vitro and in human cells. Furthermore, subcellular localization studies showed that Hippi and Apoptin perfectly colocalized in the cytoplasm of normal human HEL cells, whereas in cancerous HeLa cells most Apoptin and Hippi were located separately in the nucleus and cytoplasm and, thus, showed only a modest colocalization. Mapping studies indicate that Hippi binds within the self multimerization domain of Apoptin, and Apoptin binds to the C-terminal half of Hippi, including its death effector domain-like motif. Our results suggest that the Apoptin-Hippi interaction may play a role in the suppression of apoptosis in normal cells. PMID- 12745084 TI - Inhibition of DNA polymerases and DNA topoisomerase II by triterpenes produced by plant callus. AB - We found that some triterpene compounds could not only selectively inhibit the activities of mammalian DNA polymerase alpha (pol alpha) and beta (pol beta), but could also potently inhibit DNA topoisomerase II (topo II) [Biochem. J. 350 (2000) 757]. Here, we report that natural triterpenes produced by callus from an ancient Chinese medicinal plant were also inhibitors of the enzymes, and some were more selective than others. The natural triterpenes with a carboxyl group equally inhibited the activities of pol alpha, pol beta, and topo II, while the olide-type triterpenes with a ketone group suppressed the activities of pol beta and topo II, but not pol alpha. The other triterpenes from the callus hardly influenced these enzyme activities. As also described previously [J. Biochem. 130 (2001) 657], pol beta and topo II have a three-dimensionally similar triterpene binding region, which is a pocket in which specific compounds can insert. The newly found triterpene inhibitors might structure-dependently insert into the pocket, and the pocket structure of each enzyme might, three-dimensionally but slightly, differ among them. The triterpene frames could be used for screening new inhibitors of the enzymes, and computer-simulated drug design using the frame and pocket structure may in theory be a possible approach to develop new inhibitors. PMID- 12745085 TI - Survivin expression in the stomach: implications for mucosal integrity and protection. AB - Survivin, an apoptosis inhibitor, is highly expressed in a majority of human cancers and is required during embryonic development. Our present studies show that survivin is also expressed in normal gastric mucosa of adult humans and rats. In both human and rat gastric mucosa, survivin is expressed predominantly in the nuclei of mucosal surface epithelial cells. In rats, survivin is also detected in the nuclei of some neck cells, whereas in the humans, survivin is expressed in both nuclei and cytoplasm of chief and parietal cells. Furthermore, survivin is expressed at higher levels in the nuclei of cultured gastric mucosal epithelial cells than in gastric microvascular endothelial cells, which supports the expression pattern in intact tissues. Based on these expression studies, and the known role of survivin as an anti-apoptosis protein, survivin may play a role in maintaining gastric mucosal integrity and regulating cell renewal in the gastric mucosa. PMID- 12745086 TI - Monocyte chemoattractant protein-1 induces scavenger receptor expression and monocyte differentiation into foam cells. AB - Accumulation of monocytes and the entrapment of oxidized-low-density lipoprotein (ox-LDL) in monocytes are important in the differentiation into "foam" macrophages and the pathogenesis of atherosclerosis. We investigated the role of monocyte chemoattractant protein-1 (MCP-1) in the expression of scavenger receptor (SCR) by using resting monocytes prepared by counterflow centrifugal elutriation. Our results showed that: (1) MCP-1 increased the expression of CD36 SCR by flow cytometric analysis. (2) MCP-1 increased incorporation of 125I labeled ox-LDL and oil red O staining. (3) MCP-1 and ox-LDL enhanced in vitro transendothelial monocyte migration. (4) These functions were mediated at least in part via extracellular signal-regulated kinase (ERK) pathway. (5) MCP-1 and ox LDL did not induce monocyte proliferation. Our results imply that MCP-1 is involved in the inflammatory process of atherosclerosis through the induction of SCR expression via the ERK pathway and differentiation of monocytes into foam macrophages, as well as induction of monocyte migration. PMID- 12745087 TI - [2-Sulfo-9-fluorenylmethoxycarbonyl]3-exendin-4-a long-acting glucose-lowering prodrug. AB - Exendin-4, a glucagon-like peptide-1 agonist, is a relatively short-lived species in the circulatory system in vivo. We have linked three moieties of 2-sulfo-9 fluorenylmethoxycarbonyl (FMS) to the three amino functions of exendin-4. FMS(3) exendin-4 thus obtained has about 0.1% the glucose-lowering potency of the native peptide. Upon in vitro incubation at physiological conditions, the FMS moieties undergo hydrolysis generating the parent fully active, exendin-4. A single subcutaneous administration of FMS(3)-exendin-4 to healthy and type II diabetic mice has induced a glucose-lowering profile that exceeds in length several times that obtained by the native peptide. The reduction of blood glucose level began 1h after administration and was maximal 3-4h later. The blood glucose level returned to pre-administered values with t(1/2) of 12+/-0.7, 26+/-2, and 44+/-3h with doses of 1, 10, and 100 micro g/mouse, respectively. In sum, we have synthesized and characterized FMS(3)-exendin-4, a prodrug derivative of the native insulinotropic peptide, and found it to facilitate a prolonged and stable, glucose-lowering action in healthy and diabetic mice. PMID- 12745088 TI - Subcellular localization of collapsin response mediator proteins to lipid rafts. AB - Collapsin response mediator proteins (CRMPs) are involved in signal transduction after exposure of neural cells to the axon guidance molecule Semaphorin 3A/collapsin. All five known CRMPs are expressed in the developing cerebral cortex and neocortical neurons are responsive to Semaphorin 3A. Here, we examine the expression and subcellular localization of CRMPs in neocortical neurons and in neonatal rat brain. In neocortical neurons CRMP-4 was detected in the perikaryon with a diffuse cytosolic distribution. In neurites and at growth cones punctate staining patterns were observed. Extraction of neuron cultures with methyl-beta-cyclodextrin to deplete cholesterol caused rapid redistribution of the punctate CRMP-4 staining into larger patches and abundant growth cone collapse. Western blotting of brain extracts demonstrated for all CRMPs the existence of soluble, detergent-extractable, and Triton X-100-resistant forms. Furthermore, sucrose density gradient centrifugation after solubilization of brain membranes with Triton X-100 revealed that CRMP-1, -3, -5, and to a lower extent CRMP-4 are associated with a detergent-resistant fraction with low buoyant density, but CRMP-2 was not detectable in this fraction. Thus, we propose that lipid rafts form sites for the compartmentalization of signaling events involving specific CRMPs and that the integrity of these membrane microdomains is essential for the maintenance of growth cones. PMID- 12745089 TI - Rat cerebral endothelial cells express trk C and are regulated by neurotrophin-3. AB - Cerebral endothelial cells (CEC) are critical for formation of the vascular system in the mammalian central nervous system (CNS). We focused on the neurotrophin (NT) for its possible involvement in signaling for the regulation of CEC to control formation and maintenance of the vascular system in CNS in comparison of rat cerebral endothelial cells (RCEC) with rat aortic endothelial cells (RAEC). We found that (1) trk C, a receptor for neurotrophin-3 (NT-3), is dominantly expressed in RCEC, but trk B, a receptor for brain-derived neurotrophic factor, is dominantly expressed in RAEC; (2) NT-3 inhibited the proliferation of RCEC; and (3) NT-3 stimulated the production of nitric oxide (NO) with increases in protein expression of endothelial NO synthase. These data indicated that NT may regulate and/or maintain the functions of the brain microvasculature through the regulation of CEC. PMID- 12745090 TI - Nearest neighbour algorithm for predicting protein subcellular location by combining functional domain composition and pseudo-amino acid composition. AB - In this paper, based on the approach by combining the "functional domain composition" [K.C. Chou, Y. D. Cai, J. Biol. Chem. 277 (2002) 45765] and the pseudo-amino acid composition [K.C. Chou, Proteins Struct. Funct. Genet. 43 (2001) 246; Correction Proteins Struct. Funct. Genet. 2044 (2001) 2060], the Nearest Neighbour Algorithm (NNA) was developed for predicting the protein subcellular location. Very high success rates were observed, suggesting that such a hybrid approach may become a useful high-throughput tool in the area of bioinformatics and proteomics. PMID- 12745091 TI - Expression and phosphorylation of the replication regulator protein geminin. AB - It has been described that the replication regulator protein geminin is rapidly degraded at the end of mitosis and newly expressed at the beginning of the next S phase in the metazoan cell cycle. We have performed experiments to investigate the synthesis of geminin in cycling human HeLa cells. The levels of geminin-mRNA vary only modestly during the cell cycle with a 2-3-fold higher mRNA level at the G1/S phase transition, whereas newly synthesized geminin can only be detected in post-G1 phases. Surprisingly, geminin, once synthesized, does not remain stable, but is turned over during S phase with a half-life of 3-4h. We also show that geminin becomes phosphorylated as S phase proceeds and identify by MALDI mass spectrometry two specific major phosphorylation sites. PMID- 12745092 TI - N-terminal extensions of the human AMPD2 polypeptide influence ATP regulation of isoform L. AB - Human tissues and cells express three AMP deaminase (AMPD) isoforms containing divergent N-terminal domains, and each member of the multigene family encoding these enzymes produces alternative transcripts that confer additional N-terminal divergence through extensions and cassette-type substitutions. Available data suggest that divergent N-terminal domains can influence AMPD isoform behavior, but the functional significance for additional divergence within each enzyme is unknown. Three isoform L (AMPD2) variants, 1A/2, 1B/2, and 1B/3, contain N terminal extensions of 47, 128, and 53 amino acids, respectively. This study has determined the kinetic and regulatory behaviors of these three isoform L enzymes in the presence of positive (ATP) and negative (phosphate) allosteric effectors. All display nearly identical kinetic parameters and regulatory responses in the presence of phosphate alone, or in combination with ATP. Regulation by ATP is biphasic and the three isoform L enzymes also exhibit similar activation profiles and maximum initial velocities at 2-3mM in the presence of 1mM phosphate, whereas higher concentrations of phosphate suppress this activation. However, maximum initial velocities are achieved at lower ATP concentrations (0.8-1.5mM) in the absence of phosphate and under these conditions 1B/2 is less active, 1B/3 is more active, and 1A/2 is similarly active when compared to 1mM phosphate over the range of ATP concentrations found in non-muscle cells (0.8-3.7mM). These combined results suggest that isoform L enzymes are designed to function under different metabolic conditions encountered in the non-striated muscle environments where they are expressed. PMID- 12745093 TI - An absolute role of the PKC-dependent NF-kappaB activation for induction of MMP-9 in hepatocellular carcinoma cells. AB - Matrix metalloproteinases (MMPs) play an important role in inflammation, tumor cell invasion, and metastasis. We found that phorbol-12-myristate-13-acetate (PMA)-stimulated invasion of the hepatocellular carcinoma (HCC) SNU-387 and SNU 398 cells and that PMA induced the secretion of MMP-9 in the cells, but did not induce the secretion of MMP-2. The PMA-induced MMP-9 secretion was abolished by treatment of a pan-protein kinase C (PKC) inhibitor, GF109203X, and an inhibitor of NF-kappaB activation, sulfasalazine, and partly inhibited by treatment of inhibitors of ERK pathway, PD98059 and U0126. In addition, the PMA-stimulated activation of the MMP-9 promoter was completely inhibited by a mutation of the NF kappaB site within the MMP-9 promoter, but not completely by mutations of two AP 1 sites. Moreover, the MMP-9 induction by HGF and TNF-alpha was also completely inhibited by GF109203X and sulfasalazine, but not by PD98059 and U0126. These data demonstrate that the PKC-dependent NF-kappaB activation is absolute for MMP 9 induction and that the PKC-dependent ERK activation devotes to increase the expression level of MMP-9, in HCC cells. PMID- 12745094 TI - The fragile X mental retardation protein interacts with U-rich RNAs in a yeast three-hybrid system. AB - We recently identified several ESTs that bind to the fragile X mental retardation protein (FMRP) in vitro. To determine whether they interacted in vivo we performed three-hybrid screens in a Saccharomyces cerevisiae histidine auxotroph. We demonstrate that two of the ESTs support growth on histidine and transduce beta-galactosidase activity when co-expressed with FMRP under selective growth conditions. In contrast, the iron response element (IRE) RNA does not. Likewise, the ESTs do not support growth or transduce beta-galactosidase activity when co expressed with the iron response element binding protein (IRP). Each EST is relatively small and has 40% identity with a sequence in FMR1 mRNA harboring FMRP binding determinants. Interestingly, while neither the ESTs contain a G-quartet structural motif they do contain U-rich sequences that are found in mRNA with demonstrated in vitro binding and in vivo association with FMRP. This indicates that U-rich elements comprise another motif recognized by FMRP. PMID- 12745095 TI - Effectiveness of theophylline prophylaxis of renal impairment after coronary angiography in patients with chronic renal insufficiency. AB - Contrast media can lead to renal impairment that results in longer hospitalization and increased mortality. Adenosine is a crucial mediator of contrast-induced nephropathy (CIN; an increase in serum creatinine of >or=0.5 mg/dl within 48 hours). Therefore, it was the purpose of our study to investigate whether the adenosine antagonist theophylline reduces the incidence of CIN after coronary angiography. We also characterized risk factors for CIN after coronary angiography. One hundred patients with serum creatinine concentrations of >or=1.3 mg/dl randomly received 200 mg IV theophylline or placebo 30 minutes before coronary angiography (amount of contrast medium >or=100 ml). Patients who received theophylline and the controls were comparable with regard to baseline creatinine levels (means +/- SD) (1.65 +/- 0.41 vs 1.72 +/- 0.69 mg/dl) and the amount of contrast medium received (235 +/- 89 vs 261 +/- 139 ml). Theophylline significantly reduced the incidence of CIN (4% vs 20%, p = 0.0138). With placebo, creatinine significantly increased at 12 (1.82 +/- 0.79 mg/dl, p = 0.0057), 24 (1.90 +/- 0.86 mg/dl, p = 0.0001), and 48 hours (1.90 +/- 0.89 mg/dl, p = 0.0007) after administration of contrast medium. With pretreatment with theophylline, mean creatinine only increased 24 hours after contrast medium administration (1.70 +/- 0.40 mg/dl, p = 0.029), but was stable 12 hours (1.65 +/- 0.43 mg/dl, p = 0.99) and 48 hours after contrast medium administration (1.65 +/- 0.41 mg/dl, p = 0.99). The following parameters were significantly associated with contrast induced renal impairment: Cigarroa quotient >5 (contrast medium [milliters] x serum creatinine/body weight [kg]), elevated troponin T, >300 ml of contrast medium, and emergency angiography. In conclusion, theophylline reduces the incidence of CIN in patients with chronic renal insufficiency undergoing coronary angiography. It should be used especially in patients receiving large amounts of contrast medium, and in patients with a Cigarroa quotient of >5 and/or elevated troponin T levels. PMID- 12745096 TI - Time for contrast material to traverse the epicardial artery and the myocardium in ST-segment elevation acute myocardial infarction versus unstable angina pectoris/non-ST-elevation acute myocardial infarction. AB - Although the time for contrast material to fill the epicardial artery in the setting of acute coronary syndromes has been studied extensively, the time for contrast material to fill the myocardium has not been evaluated. We compared differences in myocardial contrast material transit among patients with unstable angina pectoris/non-ST-elevation acute myocardial infarction (UAP/NSTEAMI) with patients with ST-elevation acute myocardial infarction (STEAMI). The time it took for contrast material to first appear and to arrive at peak intensity in the myocardium was compared in 224 patients with STEAMI enrolled in the LIMIT-AMI study versus 430 patients with UAP/NSTEAMI enrolled in the TACTICS-TIMI 18 trial. In patients with STEAMI, there was a delay in both the time for contrast material to first enter the myocardium (5,619 +/- 1,789 vs 4,663 +/- 1,626 ms, p <0.0001) and the time from entrance to peak blush intensity (2,387 +/- 1,359 vs 1,959 +/- 1,244 ms, p = 0.003) compared with patients with UAP/NSTEAMI. STEAMI remained significantly associated with impaired entrance of contrast material into the myocardium (p <0.0001) in a multivariate model controlling for known correlates of impaired epicardial flow (presence of thrombus, percent diameter stenosis, left anterior descending artery location, and contrast material inflow in the epicardial artery [corrected TIMI frame count]). The time for contrast material to enter the myocardium is impaired to a greater degree in STEAMI compared with UAP/NSTEAMI, even after adjusting for other variables known to delay flow in the epicardial artery. These data provide insight into potential mechanistic differences between these 2 clinical syndromes. PMID- 12745097 TI - Comparison of in-hospital and one-year outcomes in patients with left ventricular ejection fractions or=50% having percutaneous coronary revascularization. AB - Outcome studies of percutaneous coronary intervention (PCI) with conventional balloon angioplasty have established increased in-hospital and 1-year mortality in patients with left ventricular (LV) dysfunction compared with others. It is unclear whether recent PCI practice innovations, including stents and adjunctive pharmacotherapy, have made PCI safer and more effective in patients with LV dysfunction. We evaluated the influence of LV ejection fraction (EF) indexes on in-hospital and 1-year outcomes in 1,458 patients within the National Heart, Lung, and Blood Institute-sponsored Dynamic Registry. Patients (n = 300) with acute myocardial infarction were excluded. The remaining 1,158 patients were subdivided into 3 categories: group 1, EF or=50% (n = 866). We determined the frequency of individual and composite adverse events (death/myocardial infarction [MI]/coronary artery bypass grafting) at discharge and 1 year. In the Dynamic Registry patients, mean EF in the 3 groups was 32%, 45%, and 62% and in-hospital mortality was 3.0%, 1.6%, and 0.1%, respectively (p <0.001). The composite end point of death/MI was also significant, but other in-hospital adverse events did not differ between groups. The respective mortality rates were 11.0%, 4.5%, and 1.9% (p <0.001) after 1 year. The composite end points of death/MI and death/MI/coronary artery bypass grafting also occurred more frequently in group 1 patients. Thus, significant LV dysfunction was still associated with increased in hospital and 1-year mortality in patients having contemporary PCI. PMID- 12745098 TI - Concordance/discordance between plasma apolipoprotein B levels and the cholesterol indexes of atherosclerotic risk. AB - The objective of the present study was to examine concordance/discordance among 4 atherogenic indexes of cardiovascular risk: plasma total cholesterol, low-density lipoprotein (LDL) cholesterol, non-high-density lipoprotein (non-HDL) cholesterol, and apolipoprotein B-100 (apoB). Analyses were conducted in a cohort of 2,103 men without coronary artery disease (CAD) at the onset of the Quebec Cardiovascular Study. Although there were strong and highly significant correlations among the 4 risk indexes (0.78 < r < 0.97), only 50% of all subjects had concordant apoB and LDL cholesterol levels (i.e., values that fell into the same quintile of the population distribution). Moreover, concordance/discordance was not the same throughout the range of both variables; it was greater at the extremes of their respective distributions (65%), but significantly less in the midpoints (<40%). ApoB appeared to be more concordant with non-HDL cholesterol than with LDL cholesterol, although >1/3 of all subjects had discordant levels. Kappa analysis confirmed that there was only fair agreement between apoB and total or LDL cholesterol (0.38 and 0.36, respectively) and only moderate agreement between non-HDL cholesterol and apoB (0.47). Finally, a significant proportion of subjects (528 of 2,103) who had disproportionately higher apoB levels than would have been predicted based on their LDL cholesterol concentrations was more obese and manifested several features of the metabolic syndrome. They also had a significantly increased cardiovascular risk. In summary, plasma apoB and the various cholesterol indexes are complementary rather than competitive indexes of atherosclerotic risk and provide further evidence as to why measurement of apoB should be part of a standard lipoprotein assessment of CAD risk. PMID- 12745099 TI - Multiple accessory pathways in pediatric patients with Wolff-Parkinson-White syndrome. AB - The characteristics of multiple accessory pathways in children have not been previously studied. Records were reviewed of 317 consecutive pediatric patients with Wolff-Parkinson-White syndrome who underwent electrophysiologic study and radiofrequency catheter ablation at our institution. Twenty-eight patients (9%) had multiple pathways (a total of 64 pathways: 21 patients had 2, 6 had 3, and 1 patient had 4 pathways). The locations were left free wall (22 pathways), right free wall (19 pathways), posteroseptal (17 pathways), and anteromidseptal (6 pathways). Of these 64 pathways, 55 were ablated successfully without complications, 5 failed ablation, and 4 fasciculoventricular fibers did not require treatment. Three patients had a newly found pathway at the repeat session. Three patients had atrioventricular nodal reentrant tachycardia and the slow pathway area was modified. One patient had failed initial ablation and had a successful ablation using a 3-dimensional electroanatomic mapping system. Compared with patients with a single pathway, those with multiple pathways exhibited a higher incidence of antidromic tachycardia, a shorter anterograde accessory pathway effective refractory period (<250 ms), a longer fluoroscopic time (65 +/- 43 vs 39 +/- 46 minutes, p <0.05), and a larger number of unsuccessful attempts (9 +/- 16 vs 5 +/- 8, p <0.05). Success rate (92% vs 93%) and recurrence rate (1.7% vs 2.1%) were similar in both groups. This study demonstrates that multiple pathways are not rare in pediatric patients and that multiple pathways contrast with a single pathway in a variety of conduction properties. PMID- 12745100 TI - Significance of bundle branch block during atrioventricular nodal reentrant tachycardia. AB - There are very limited data on the effects of bundle branch block (BBB) in patients with atrioventricular nodal reentrant tachycardia (AVNRT). Studies in a total of 155 patients with 162 episodes of AVNRT were retrospectively analyzed. A total of 38 patients (25%) developed spontaneous right BBB, whereas 5 (3%) developed left BBB during tachycardia. Five of the 38 (13%) with right BBB showed near identical prolongation of both the ventriculoatrial (VA) (15 +/- 5 ms; 10 to 23) and His to atrial intervals (HA) (14 +/- 4 ms; 10 to 20) with an identical atrial activation sequence for both right BBB or normal QRS tachycardia complexes. In contrast, all 5 patients with left BBB showed a decrease in the VA (-18 +/- 11 ms; 10 to 36) with unchanged HA comparing left BBB to normal QRS patterns during AVNRT. The magnitude of prolongation of the His to ventricular interval (HV) during left BBB (19 +/- 12 ms; 10 to 40) was nearly identical to the decrease in the VA. In conclusion, prolongation of VA and HA with unchanged HV in patients with AVNRT and right BBB suggests that right BBB is due to a block in the fibers in close proximity to the His recording site. The data suggest that fibers in the His bundle are predestined to activate the right bundle branch, and in AVNRT the lower turnaround point may be within the His bundle. PMID- 12745101 TI - Relation of cardiac sympathetic innervation to proinflammatory cytokine levels in patients with heart failure secondary to idiopathic dilated cardiomyopathy. AB - Experimental studies have shown that cytokine production by the heart may be regulated by sympathetic nervous system stimulation of cardiac beta-adrenergic receptors. Proinflammatory cytokine levels are increased in heart failure, whereas cardiac fixation of 123-I-metaiodobenzylguanidine (MIBG) has been used to study myocardial adrenergic innervation. This study was designed to assess the relation between cardiac MIBG uptake and circulating levels of proinflammatory cytokines in patients with idiopathic dilated cardiomyopathy (IDC). Forty-seven patients (12 women; mean age 56.5 +/- 9 years) with angiographically proved IDC, in New York Heart Association functional classes II to III, and who had left ventricular ejection fraction 30.6 +/- 9.5%, and 20 healthy controls were studied with planar MIBG. The early (10 minutes) and late (4 hours) heart to mediastinum uptake ratio and washout were calculated. Circulating plasma levels of interleukins (IL)-1 and IL-6, tumor necrosis factor-alpha, and solube receptors of TNF (sTNFR) I and II were measured. The patient group had significantly lower values of MIBG uptake at 10 minutes (p <0.001) and 4 hours (p <0.001) and higher washout (p <0.001) than the controls. Late MIBG uptake was significantly correlated with New York Heart Association class (r = -0.42, p = 0.02), left ventricular ejection fraction (r = 0.34, p = 0.01), left ventricular systolic wall stress (r = -0.40, p = 0.05), oxygen consumption at peak exercise (r = 0.32, p = 0.03), IL-1 (r = -0.55, p <0.001), TNF (r = -0.33, p = 0.02), and sTNFRII (r = -0.44, p = 0.001). Multivariate linear regression analysis revealed that MIBG at 4 hours was independently associated with IL-1 levels (p <0.001). Thus, reduced cardiac sympathetic innervation in heart failure is associated with elevated levels of inflammatory cytokines, suggesting that it has a potential inflammatory effect via modulation of the cardiac production of these cytokines. PMID- 12745102 TI - Relation of left ventricular thickness to age and gender in hypertrophic cardiomyopathy. AB - Left ventricular (LV) wall thickening is the most consistent clinical marker of hypertrophic cardiomyopathy (HC), and characteristically increases substantially during adolescence. In this study, we used 2-dimensional echocardiography to develop a cross-sectional profile of LV wall thicknesses in adult patients with HC. We studied a regional community-based cohort of 239 consecutively enrolled patients (aged 18 to 91 years). On average, maximum LV wall thickness decreased relative to increasing age (p = 0.007) within 4 age groups: 22.8 +/- 5.1 mm (18 to 39 years) to 22.1 +/- 5.1 mm (40 to 59 years) to 21.1 +/- 3.7 mm (60 to 74 years) to 20.8 +/- 3.6 mm (>or=75 years). The LV thickness index (summation of wall thicknesses in all 4 segments) also decreased with age (p = 0.017): 63.0 +/- 12.2 mm to 59.8 +/- 11.9 mm to 58.3 +/- 10.4 mm to 57.9 +/- 9.8 mm. Decreasing magnitude of LV hypertrophy was independently associated with increasing age, but not with other relevant disease variables, such as symptoms and outflow obstruction. However, when separated by gender, this inverse relation between age and LV wall thickness was statistically significant only for women (p = 0.007). In conclusion, in an unselected HC cohort, cross-sectional analysis showed a modest but statistically significant inverse relation between age and LV hypertrophy that was largely gender-specific for women. This association constitutes another facet of the natural history of this complex and heterogenous disease and may reflect disproportionate occurrence of premature death in young patients with HC with marked hypertrophy or possibly gradual LV remodeling. PMID- 12745104 TI - Nanette Kass Wenger, MD: a conversation with the editor [interviewed by William Clifford Roberts]. PMID- 12745103 TI - The dark side of platelet glycoprotein IIb/IIIa receptor inhibitors during percutaneous coronary interventions. PMID- 12745105 TI - Comparison of prevalence and severity of coronary calcium determined by electron beam tomography among various ethnic groups. PMID- 12745106 TI - Comparison of 30-day outcome, resource use, and coronary artery disease severity in patients with suspected coronary artery disease with and without diabetes mellitus assigned to chest pain units. PMID- 12745107 TI - Comparative effects of diet and simvastatin on markers of thrombogenicity in patients with coronary artery disease. PMID- 12745108 TI - Effect of single dose of intravenous heparin on plasma levels of angiogenic growth factors. PMID- 12745109 TI - Effect of intravenous insulin administration on left ventricular performance during non-ST-elevation acute coronary events in patients with diabetes mellitus. PMID- 12745111 TI - Role of factor XIII Val34Leu polymorphism in patients <45 years of age with acute myocardial infarction. PMID- 12745110 TI - Effect of short-term estrogen with and without progesterone therapy on circulating markers of endothelial activation and injury in postmenopausal women with unstable angina pectoris. PMID- 12745112 TI - Comparison of supine bicycle exercise and treadmill exercise Doppler echocardiography in evaluation of patients with coronary artery disease. PMID- 12745113 TI - Usefulness of an acute coronary syndrome pathway to improve adherence to secondary prevention guidelines. PMID- 12745114 TI - One-year survival in patients with acute myocardial infarction and a saphenous vein graft culprit treated with primary angioplasty. PMID- 12745115 TI - Comparison of risk profiles and outcomes in women versus men >or=75 years of age undergoing coronary artery bypass grafting. PMID- 12745116 TI - Usefulness of circulating interleukin-18 concentration in acute myocardial infarction as a risk factor for late restenosis after emergency coronary angioplasty. PMID- 12745117 TI - Implications of the presence and length of "geographic miss" on restenosis and the edge phenomenon in the INHIBIT trial. PMID- 12745119 TI - Influence of various percutaneous coronary interventional devices on postinterventional luminal shape and plaque surface characteristics as determined by intravascular ultrasound. PMID- 12745118 TI - Effect of homocysteine-lowering therapy on restenosis after percutaneous coronary intervention for narrowings in small coronary arteries. PMID- 12745120 TI - Prognostic implication of troponin I elevation after percutaneous coronary intervention. PMID- 12745121 TI - Hormone replacement therapy and risk of acute myocardial infarction in postmenopausal women with diabetes mellitus. PMID- 12745122 TI - Usefulness of echocardiography in infants with supraventricular tachycardia. PMID- 12745123 TI - Effect of nortriptyline on the day-night systolic blood pressure difference in hypertensive and normotensive elderly depressed women. PMID- 12745124 TI - Effect of carvedilol on exercise tolerance in patients with chronic heart failure and a restrictive left ventricular filling pattern. PMID- 12745125 TI - Effect of atenolol therapy on left atrial appendage function in patients with symptomatic mitral stenosis. PMID- 12745126 TI - Endothelial function following the Fontan operation. PMID- 12745128 TI - Munchausen syndrome presenting as cardiovascular disease (cardiopathia fantastica). PMID- 12745127 TI - Taming of the shrew. PMID- 12745130 TI - Significance of QT dispersion. PMID- 12745129 TI - Why did green tea not protect against coronary artery disease but protect against myocardial infarction? PMID- 12745131 TI - Blood pressure control and rates of edema following the administration of the cyclooxygenase-2 specific inhibitors celecoxib versus rofecoxib in patients with systemic hypertension and osteoarthritis. PMID- 12745132 TI - Action starts now to control disease due to schistosomiasis and soil-transmitted helminthiasis. PMID- 12745133 TI - Quantification of clinical morbidity associated with schistosome infection in sub Saharan Africa. AB - Health policy making in developing countries requires estimates of the (global) burden of disease. At present, most of the available data on schistosomiasis is limited to numbers of individuals harbouring the infection. We explored the relationship between the presence of schistosome infection and clinical morbidity, in order to estimate numbers of individuals with disease-specific morbidity for Schistosoma haematobium and Schistosoma mansoni infection in sub Saharan Africa. We searched the literature for cross-sectional data from field studies reporting both schistosome infection and morbidity. This was used to derive a functional relationship between morbidity and infection. After standardisation for diagnostic method, the number of individuals with specific types of clinical morbidity or pathology was predicted. As only aggregated prevalences of infection were available for countries or areas, we adjusted for heterogeneity in infection levels within communities in those countries. In total, 70 million individuals out of 682 million (2000 estimate) in sub-Saharan Africa were estimated to experience haematuria in the last 2 weeks associated with S. haematobium infection, and 32 million dysuria. Ultrasound detected serious consequences of S. haematobium, major bladder wall pathology and major hydronephrosis, were predicted at 18 and 10 million, respectively. Infection with S. mansoni was estimated to cause diarrhoea in 0.78 million individuals, blood in stool in 4.4 million and hepatomegaly in 8.5 million. As the associations between prevalence of S. mansoni infection and prevalence of diarrhoea and blood in stool were not very clear, the resulting estimates may be underestimations. Using the very limited data available, we estimated the mortality rates due to non functioning kidney (from S. haematobium) and haematemesis (from S. mansoni) at 150000 and 130000 per year. Given the overall high number of cases with schistosomiasis-related disease and associated death, we conclude that schistosomiasis remains an important public health problem in sub-Saharan Africa. PMID- 12745134 TI - Albendazole, mebendazole and praziquantel. Review of non-clinical toxicity and pharmacokinetics. AB - The pharmacokinetics and toxicity of albendazole, mebendazole and praziquantel are extensively reviewed, drawing on original published work and reviews in the open scientific literature and on assessments by international agencies and official regulatory bodies in Europe and the USA. Information about human and veterinary medical uses and adverse reactions is evaluated. The totality of the non-clinical information available about these long-established drugs may not comply with current official guidelines for new medicines but reasons are given why the "deficiencies" are only apparent and the data gaps can be replaced by other results, largely obtained from the target species and the many years of clinical experience of safe use of these drugs in humans and animals. PMID- 12745135 TI - The impact of chemotherapy on morbidity due to schistosomiasis. AB - Current knowledge on the impact of chemotherapy on schistosomiasis-related morbidity is still fragmentary. In urinary schistosomiasis, reversal of organ pathology follows cure after 6 months and resurgence takes place after at least another 6 months. Retreatment after less than 1 year is, therefore, unnecessary. Also, intestinal schistosomiasis appears to regress promptly after chemotherapy. For the reversal of hepatic morbidity, more than one chemotherapy round appears necessary at least in foci of intense transmission of schistosomiasis. The earlier chemotherapy is given, the higher the chances of reversal of schistosomal pathology, but pathology may regress to some extent also in adults. The regression and resurgence of periportal fibrosis, as detected by ultrasonography, occurs with a delay of 7 months to more than 2 years after therapy. Retreatment after less than 1 year may not permit full assessment of the impact of the first round on hepatic morbidity. Children and adolescents should be the major target population, taking into account that in many foci, children out-of-school must be covered because they are at the highest risk. Repeated treatment during childhood may prevent the development of urinary tract disease in adulthood. However, no data are available on the prevention of genital pathology. Repeated chemotherapy may have a long term effect on re-infection intensities and the development of severe morbidity, even in foci where control has been interrupted for many years. Severe hepatic fibrosis may be prevented even in foci of intense transmission provided more than two rounds of chemotherapy have been given in childhood and that chemotherapy is available on demand. Chemotherapy has an important impact on child development, physical fitness and working capacity. Its effect on growth and anemia is improved by simultaneous treatment of intestinal parasites and the provision of adequate iron supplementation. The impact of chemotherapy on many of the multifaceted manifestations of schistosomiasis has not been assessed systematically. More data are needed on gallbladder pathology, neuroschistosomiasis, endocrinologic disorders, bladder cancer and co-infections with other pathogens. In areas where control has been achieved, the overall morbidity and mortality has decreased with a delay of many years or even decades. PMID- 12745136 TI - Administration of praziquantel to pregnant and lactating women. AB - Praziquantel (PZQ) is the safest of all anti-helminthics and now forms the backbone for all national control programs against schistosomiasis (Med. Res. Rev. 3 (1983) 147-200; Bull. WHO 57 (1979) 767-771; Wegner, D.H.G, Therapeutic Drugs (1991), Churchill Livingstone; Adv. Intern. Med. 32 (1987) 193-206; Drugs 42 (1991) 379-405; Pharmac. Ther. 68 (1995) 35-85; Ann. Intern. Med. 110 (1989) 290-296). Despite its lack of known toxicity, the drug was not tested on pregnant or lactating women prior to release. It is currently listed as Pregnancy Category B by the US FDA, which is a drug presumed safe based in animal studies. Unfortunately, this has been interpreted by most national control programs and WHO (1998) to exclude lactating and pregnant women from treatment. In fact, some experts advocate excluding adolescent girls from mass treatment campaigns over this issue. As a result, a large number of women living in endemic countries are currently left untreated or have treatment significantly delayed. A review of the current known toxicology of PZQ, combined with over two decades of clinical experience with this drug, suggest very low potential for adverse effects on either the mother or her unborn child. In contrast, significant animal and human data are presented in this review that suggest both the pregnant woman and her unborn fetus suffer morbid sequella from schistosomiasis. A double-blind placebo controlled trial that could resolve this issue would require a very large and expensive study and in light of the above facts might not now be ethically appropriate. The author concludes that pregnant women should be treated with PZQ, that women of childbearing age should be included in all mass treatment programs and that lactating women are not systematically excluded from treatment. PMID- 12745137 TI - Impact of mass chemotherapy on the morbidity due to soil-transmitted nematodes. AB - This review summarises current knowledge of the ill-effects of soil-transmitted helminthiasis and takes a detailed look at studies that have been published over the past decade describing the effect of mass anthelminthic use on the health of endemic communities. Mass chemotherapy appears to give maximal returns in terms of improved health in areas where hookworm is a major problem and albendazole is used regularly, along with iron supplements; in children it improves physical growth and iron stores, and in pregnant women it reduces the prevalence of iron deficiency anaemia. In areas where ascariasis is common, the directly attributable benefits of chemotherapy may be minimal, but it can facilitate the entry of other health care programmes in children, because deworming for ascariasis is often much desired and appreciated by the community. In areas with Vitamin A deficiency and endemic ascariasis, Vitamin A supplementation can be combined with deworming: anthelminthics do not impair Vitamin A absorption but the worms may interfere with Vitamin A uptake by reducing fat absorption. Where trichuriasis is a major problem, single dose chemotherapy may take some time to reduce prevalence, but reduction of heavy infections will reduce the incidence of Trichuris Dysentery Syndrome, probably benefit the learning abilities of affected schoolchildren, and may reduce anaemia and stunting. In general, children should be treated as early as possible, and in areas of very high prevalence, thrice yearly mass chemotherapy probably improves health better than twice-yearly treatment. PMID- 12745138 TI - Anthelminthic drug safety and drug administration in the control of soil transmitted helminthiasis in community campaigns. AB - Helminth infections are now recognised as being a major health priority worldwide. Morbidity due to these infections can be controlled at a reasonable cost by means of periodic chemotherapy using effective drugs. Deworming campaigns targeted at high risk groups, such as school-age children, pre-school children and women of child-bearing age, are the mainstay of the control strategy launched by WHO. Anthelminthic drugs can be delivered effectively through the school system, women's associations or other community-based interventions, each of which often lack health personnel supervision. The safety of anthelminthic drugs is, therefore, of paramount importance and side effects have to be recognised and monitored, especially when generic drugs are widespread. Four anthelminthic drugs are considered to provide appropriate single dose treatment against soil transmitted helminthiasis: albendazole, levamisole, mebendazole and pyrantel. Side effects, at the dosage recommended for deworming, have been described as negligible and self-limiting. However, a limited number of reports have associated more severe adverse reactions to the distribution of anthelminthic medicines. Even if the available information cannot confirm a cause-effect relationship, it is essential that these effects are known. Ministries of Health can then set up efficient and safe delivery, monitoring and referral systems, in order to minimise the risk and maximise the benefit of periodic anthelminthic chemotherapy in communities where soil-transmitted helminthiasis is endemic. PMID- 12745140 TI - Methods to sustain drug efficacy in helminth control programmes. AB - Assessment of the efficacy of anthelminthic treatment in public health is a broad concept, which goes beyond parasitological methods and should be clearly defined according to several indicators of morbidity. Several factors may influence the efficacy of anthelminthic drugs. The quality of drug is an issue of great importance, especially when produced locally as a generic product and used in large-scale chemotherapy-based control programmes. Other factors include the drug patient interaction, the host-parasite relationship, the diagnostic method used, genetic variations between parasite strains and induced drug resistance. Veterinary scientists have warned that drug resistance can be selected through frequent mass treatment of sheep and goats and have developed a body of knowledge on evaluation of efficacy and detection of resistance in nematodes of veterinary importance. In soil-transmitted nematodes infections of humans, the egg reduction rate (ERR), the egg hatch assay (EHA) and novel molecular biological techniques may be used to monitor drug efficacy in helminth control programmes and to detect early occurrence of resistance. Evidence of reduced drug efficacy of some anthelminthics has been suggested by recent studies and strategies to prevent or delay the emergence of drug resistance in human soil-transmitted nematodes. PMID- 12745139 TI - Use of benzimidazoles in children younger than 24 months for the treatment of soil-transmitted helminthiasis. AB - Considerable experience and limited quantitative evidence indicate that infections with the soil-transmitted helminths Ascaris lumbricoides and Trichuris trichiura usually start to become established in children aged 12 months and older. Since children living in countries where the infections are endemic are at risk of morbidity, even those as young as 12 months may need to be considered for inclusion in public health programmes designed to reduce morbidity by means of regular anthelminthic chemotherapy. This situation raises the question as to whether such young children should be given anthelminthic drugs. Systems for the absorption, distribution, metabolism and elimination of drugs do not fully develop until children are in their second year of life. Current knowledge, however, reveals that the incidence of side effects linked to benzimidazole drugs in young children is likely to be the same as in older children. Accordingly, we conclude that albendazole and mebendazole may be used to treat children as young as 12 months if local circumstances show that relief from ascariasis and trichuriasis is justified. PMID- 12745142 TI - Experience with school-based interventions against soil-transmitted helminths and extension of coverage to non-enrolled children. AB - This paper reviews the experience with school-based interventions against soil transmitted helminths with regard to reduction in prevalence, intensity of infection and morbidity. It also examines the existing experience with coverage of school-based programmes to non-enrolled children. However, as this experience is limited, the paper also seeks to give an overview of the need for school control programmes to include other segments of the community. The experiences from the programmes indicate that treatment should be performed twice or thrice yearly without prior diagnosis, should be school-based and involving schoolteachers assisted by health staff, if possible. The drugs of choice are a single dose of 400 mg albendazole or 500 mg mebendazole. If intensities of Trichuris trichiura or hookworm infections are high, a double or triple dose of one of these drugs could be considered to maximise reduction in intensities. For the benefit of growth and iron status, it should be considered to supplement with iron and other micronutrients. School-based programmes should include non enrolled school age children and pre-school children, and the system of having 'treatment days' at school, where these groups are invited for treatment, seems to be a promising strategy. While antenatal clinics have been involved in the anthelminthic treatment of pregnant women, they have not covered non-pregnant adolescent girls and women. These could be offered treatment through the 'treatment days' at school mentioned earlier. PMID- 12745141 TI - Treatment and re-treatment strategies for schistosomiasis control in different epidemiological settings: a review of 10 years' experiences. AB - This paper reviews 10 years' experience with schistosomiasis control in different endemic settings in Africa, Asia and the Americas. Research projects, pilot programs and long-term large-scale programs with the objective of controlling morbidity have been included in the review. Major advances in diagnostic tools and rapid assessment techniques have evolved during the decade making it possible to follow changes in pathology after treatment and to get baseline epidemiological information at very low cost. At the same time prices of drugs like praziquantel has declined dramatically (to 80 beats/min were characterized, in comparison with those with lower HR, by reduced carotid SBP, PP, and augmentation index, resulting in a significant increase in PP amplification. In men but not in women, this pattern was associated with higher values of brachial SBP and DBP and by higher incidences of elevated glycemia and atherosclerotic alterations. In the male population, PP amplifications was, independent of HR, associated with the presence of beta blocking agents (negative association) and elevated plasma glucose. CONCLUSIONS: Hypertensive men and women with high HR have significant PP amplifications, principally because of reduced central SBP and disturbed wave reflections. beta-blocking agents and plasma glucose independently alter PP amplification in men but not in women. Whether these opposite patterns influence the gender difference in cardiovascular risk should be prospectively studied. PMID- 12745196 TI - Arterial compliance: a diagnostic marker for atherosclerotic plaque burden? AB - BACKGROUND: Previous studies have shown atherogenesis to be related with increased vessel stiffness. Measures of the arterial compliance can be performed noninvasively from pressure pulse contour analysis of arterial waveforms. In this prospective study we aimed to analyze to what extent vessel compliance can reflect the angiographic coronary artery status. METHODS: Large and small arterial elasticity indices (LAEI in milliliters per mm Hg x 10 and SAEI in milliliters per mm Hg x 100) were measured in 151 patients on the radial artery with the PulseWave Sensor HDI device. All patients were classified into diffuse coronary artery disease (CAD) (defined as stenosis length >15 mm), focal-CAD (defined as stenosis length between 1 and 15 mm), or no-CAD. RESULTS: We found both LAEI and SAEI to be reduced in the diabetic group (LAEI: 11.2 +/- 2.9 v 13.4 +/- 4.5, P =.006; SAEI: 3.7 +/- 1.6 v 4.7 +/- 2.4, P =.01). Inverse association was seen between age and LAEI (r = -0.41; P <.001) and SAEI (r = -0.38; P <.001). No-CAD was found in 31 patients, focal-CAD in 64 patients, and diffuse-CAD in 56 patients. Mean LAEI were 13.8 +/- 3.5, 13.7 +/- 4.7, and 11.3 +/- 3.5 in the groups no-CAD, focal-CAD, and diffuse-CAD, respectively (P =.004), (no-CAD versus diffuse-CAD: P =.04; focal-CAD versus diffuse-CAD: P =.009). Respective SAEI values were 5.6 +/- 2.5, 5.0 +/- 2.1, and 3.1 +/- 1.6 (P <.001), (no-CAD versus diffuse-CAD: P <.001; focal-CAD versus diffuse-CAD: P <.001). Multivariate analysis revealed SAEI (P <.001), hypercholesterolemia (P =.005), systolic blood pressure (BP) (P <.001), mean arterial BP (P <.001), pulse pressure (P =.003), and male gender (P =.001) to be diagnostic markers of the type of vessel disease. CONCLUSIONS: Compliance measurements may be used for identification of patients with diffuse atherosclerotic processes of the coronary arteries. PMID- 12745198 TI - Evaluation of blood pressure and baroreflex sensitivity by radial artery tonometry versus finger arteriolar photoplethysmography. AB - BACKGROUND: Published normative data of noninvasive blood pressures (BPs) and autonomic modulations have been primarily derived from the finger arteriole using the Finapres (Ohmeda Co., Englewood, CO), a device that is no longer manufactured. Currently, beat-to-beat BP are obtained from the radial artery using the Colin tonometer. METHODS: We compared BP and autonomic parameters in a crossover design between the two devices in 29 subjects during seated rest and a 0.1-Hz breathing protocol. In addition, we tested whether finger arteriolar BP differences were due to pressure changes exerted by the radial tonometer. RESULTS: Uniformly, BP measured at the radial artery were significantly higher than those from the finger arteriole. Radial BP (106 +/- 19.5 mm Hg) were higher than finger arteriolar BP (95.8 +/- 13.7 mm Hg) (P <.005). Tonometric baroreflex sensitivity (BRS) (24.0 +/- 18 msec/mm Hg) was higher compared to photoplethysmographic BRS (12.0 +/- 7.7 msec/mm Hg; P <.0003). Systolic BP (radial artery) (115 +/- 25 mm Hg) were higher compared to finger arteriolar BP (97.7 +/- 19 mm Hg; P <.0025) during breathing, as was BRS (25.9 +/- 11.6 msec/mm Hg v 21.5 +/- 11.6 msec/mm Hg; P <.05). Differences in the low frequency systolic BP (LF(SBP)), representative of sympathetic vasomotor modulation, between the two methods, whether absolute, normalized, or log-transformed were not observed. CONCLUSIONS: There were no differences in arteriolar BP values in the presence or absence of radial artery tonometric pressure. These findings indicate that differences exist in systolic BP and BRS using the tonometer (radial artery) versus the Finapres (Ohmeda Co.) (finger arteriole). Furthermore, these differences are not due to pressure exerted by the radial artery tonometer that supplies blood to the finger arteriole. PMID- 12745199 TI - Arterial stiffness and blood pressure self-measurement with loaned equipment. AB - BACKGROUND: A cross-sectional study was carried out in 415 hypertensive and normotensive subjects to determine the correlation between loaned self measurement blood pressure (LSEM), arterial stiffness, and the different factors that contribute to it. METHODS: The LSEM model consists of lending a number of sphygmomanometers, property of the clinic, to patients for 3-day periods. Arterial stiffness was evaluated using the carotid-to-femoral pulse wave velocity (PWV), using an automatic apparatus. To determine the importance of each of these factors, a multiple linear regression analysis was carried out. RESULTS: Of the total number of patients, 78% were women, the average age was 57 +/- 12 years, 55.8% were hypertensive, and 38.8% were diabetic. The PWV average for the whole group was 12.1 +/- 4.2 m/sec. The correlation coefficients between the PVW and the self-measurements were 0.49 (P <.001) for the pulse pressure, and 0.46 (P <.001) for the systolic blood pressure (BP), respectively. Both represented 13.0 % of the total variation. The diastolic BP obtained by self-measurement and the serum creatinine values also had an effect on the stiffness, with 2.3 % (P =.05) each one. CONCLUSIONS: The pulse pressure readings with self-measurement correlate better with the arterial stiffness, compared with the readings taken in the office. PMID- 12745200 TI - Antihypertensive therapy with verapamil SR plus trandolapril versus atenolol plus chlorthalidone on glycemic control. AB - BACKGROUND: There is evidence that diuretics and beta blockers impair glucose tolerance, whereas calcium channel blockers and angiotensin converting enzyme blockers lack this metabolic effect. We compared the effect of a combination therapy with a nondihydropyridine calcium channel blocker plus an angiotensin converting enzyme inhibitor and a beta blocker plus a diuretic on hemoglobin A(1c) (Hb A(1c)) in patients with type 2 diabetes and mild-to- moderate hypertension. METHODS: A total of 463 hypertensive outpatients with non-insulin treated type 2 diabetes on stable antidiabetic therapy for at least 3 months and with HbA(1c) between 6.5% and 10% were recruited. In a randomized, double blind trial patients were treated for 20 weeks with fixed combinations of verapamil sustained release (SR) plus trandolapril and of atenolol plus chlorthalidone following a 2-week placebo run-in period. The main outcome measures were HbA(1c), fasting plasma glucose, and fructosamine levels as well as systolic and diastolic blood pressure. RESULTS: HbA(1c) remained stable at 7.9% after administration of verapamil SR plus trandolapril and increased from 7.8% to 8.6% with atenolol plus chlorthalidone; the differences between treatment groups were significant at 4, 12, and 20 weeks of treatment and at last visit (P <.0001). Mean blood pressure fell from 169/96 to 150/85 and from 168/95 to 145/83 mm Hg after administration of verapamil SR plus trandolapril and atenolol plus chlorthalidone, respectively. Both combinations were well tolerated. CONCLUSIONS: HbA(1c) and other parameters of short- and long-term glycemic control were in a more favorable range after antihypertensive treatment with verapamil SR plus trandolapril as compared with atenolol plus chlorthalidone. PMID- 12745201 TI - Effect of losartan on oxidative stress-induced hypertension in Sprague-Dawley rats. AB - BACKGROUND: Hypertension induced by oxidative stress has been demonstrated in normal rats. In the current study, we investigated the effect of the oral AT(1) receptor blocker losartan (10 mmol/kg/day) on oxidative stress, induced by glutathione (GSH) depletion (using buthionine-sulfoximine, BSO, 30 mmol/L/day in the drinking water), in Sprague-Dawley rats. METHODS: Mean arterial pressure (MAP) was measured by tail-cuff plethysmography and the plasma levels of total 8 isoprostane, nitric oxide, prostacyclin, thromboxane A(2), angiotensin II, aldosterone, and aortic cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) were determined by enzyme immunoassay. Plasma, heart, and kidney GSH were analyzed by high-performance liquid chromatography. Aortic and renal superoxide production was determined by fluorescence spectrometry. RESULTS: In the BSO-treated group, MAP, angiotensin II, isoprostane, thromboxane A(2), and superoxide were elevated; whereas prostacyclin, GSH, cAMP, and cGMP were reduced, compared to control. Losartan alone reduced MAP, and increased renal GSH, plasma nitric oxide, angiotensin II, aldosterone, and aortic cGMP. When administered concurrently with BSO, losartan reversed the BSO-induced elevation of MAP, superoxide, and thromboxane A(2) as well as the reduction in prostacyclin and aortic cAMP levels, but did not significantly alter the reduction in GSH or the elevation in angiotensin II and aldosterone. CONCLUSIONS: Losartan attenuates BSO-induced hypertension, which appears to be mediated, in part, by angiotensin II and the prostanoid endothelium derived factors. PMID- 12745202 TI - Altered insulin-like growth factor-1 and nitric oxide sensitivities in hypertension contribute to vascular hyperplasia. AB - Vascular medial thickening, a hallmark of hypertension, is associated with vascular smooth muscle cell (VSMC) hypertrophy and hyperplasia. Although the precise mechanisms responsible are elusive, we have shown that strain induced regulation of autocrine insulin-like growth factor-1 (IGF-1) and nitric oxide (NO) reciprocally modulate VSMC proliferation. Therefore, we investigated potential IGF-1 and NO abnormalities in young (10-week-old) spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) and their respective VSMC ex vivo. The SHR had increased mean arterial pressure (173 +/- 2 v 128 +/- 3 mm Hg, n = 24, P <.05) but similar pulse pressures (31 +/- 2 v 30 +/- 3 mm Hg; P >.05) v WKY. The SHR exhibited increased aortic wall thickness in comparison with WKY (523 +/- 16 v 355 +/- 17 micro m; P <.05). No differences were seen in plasma combined NO(2) and NO(3) (NO(x)) (0.48 +/- 0.11 mmol/L for WKY v 0.58 +/- 0.18 mmol/L for SHR) or plasma IGF-1 (1007 +/- 28 ng/mL for WKY v 953 +/- 26 ng/mL for SHR). Aortic VSMC from SHR displayed enhanced proliferation in comparison with WKY (P <.05). Underlying this enhanced proliferation was altered SHR VSMC sensitivity to the antiproliferative NO donor 2,2"[Hydroxynitrosohydrazono] bis ethanimine (DETA-NO) (ID(50): 270 +/- 20 mmol/L for SHR; 150 +/- 11 mmol/L for WKY; P <.05). Basal cyclic guanosine monophosphate (cGMP) secretion from SHR VSMC was 65-fold greater than that seen from WKY (P <.001). In response to DETA-NO, cGMP secretion from SHR VSMC increased modestly (1.5-fold; P <.01), whereas treatment of WKY VSMC resulted in a 26-fold (P <.001) increase in cGMP. The SHR VSMC did not respond to exogenous IGF-1, whereas WKY VSMC exhibited a dose dependent increase in proliferation with IGF-1 (10(-10) to 10(-7) mol/L). These data suggest that VSMC hyperplasia in early hypertension is not reflected by imbalances in plasma IGF-1 or NO. Rather, altered SHR VSMC sensitivity to NO is likely responsible in part for the observed hyperproliferation seen in early stages of hypertension. PMID- 12745204 TI - Relevance of the plasma renin hormonal control system that regulates blood pressure and sodium balance for correctly treating hypertension and for evaluating ALLHAT. PMID- 12745203 TI - Placental 11beta-hydroxysteroid dehydrogenase in Dahl and spontaneously hypertensive rats. AB - Studies in normotensive rats showed that excessive fetal exposure to maternal glucocorticoids retards growth and programs hypertension in later life. This excessive exposure is proposed to occur due to a reduction of the placental barrier to maternal glucocorticoids that is provided by 11beta-hydroxysteroid dehydrogenase (11betaHSD). To assess the possible alterations of glucocorticoid placental barrier in two genetic models of hypertension - spontaneously hypertensive (SHR) and Dahl salt-sensitive rats (DS) and their normotensive counterparts Wistar-Kyoto (WKY) and Dahl salt-resistant rats (DR)-we performed real-time reverse transcriptase-polymerase chain reaction analysis and bioactivity measurements of placental 11betaHSD in the last third of gestation. Whereas 11betaHSD2 mRNA expression was not different among the investigated strains, 11betaHSD1 mRNA abundance was 2.4 times higher in WKY than in SHR and 9.6 times higher in DS than in DR placentae. The 11betaHSD2 activity studies performed in placental homogenates revealed activity that did not differ among the strains. Concomitant with 11betaHSD1 mRNA expression 11-oxoreductase activity was clearly evident in all strains and was higher in WKY and DS rats than in SHR and DR, respectively. Nevertheless, the net 11betaHSD activity of tissue fragments (11beta-dehydrogenase minus 11-oxoreductase) was tended toward dehydrogenase action, ie, toward corticosterone inactivation and was significantly lower in DS than in DR rats. The 11beta-dehydrogenase/11 oxoreductase ratio was less than 2:1 in SHR and WKY rats, whereas this ratio was 9:1 in DR and 4.5:1 in DS rats. These data suggest that the placental glucocorticoid barrier is not decreased in SHR rats in comparison with normotensive WKY but is lower in DS than in DR counterparts. It cannot be excluded, therefore, that the placental glucocorticoid barrier in Dahl rats influences the pathways that might lead to the sensitivity of blood pressure to high salt intake in later life. PMID- 12745205 TI - A specialist in clinical hypertension critiques ALLHAT. PMID- 12745206 TI - Why we can't translate clinical trials into clinical practice in hypertension. PMID- 12745207 TI - Squatting: the hemodynamic change is induced by enhanced aortic wave reflection. PMID- 12745209 TI - Cardiac angiotensin II: an intracrine hormone? PMID- 12745210 TI - Focus on noncovalent interactions. PMID- 12745211 TI - Comparative ESI-MS study of approximately 2.2 MDa native hemocyanins from deep sea and shore crabs: from protein oligomeric state to biotope. AB - In the past years, the potential of electrospray ionization mass spectrometry (ESI-MS) for the observation of intact weak interactions, such as non-covalent protein-ligand, protein-protein, protein-DNA complexes, has spread out. The coupling of ESI with time-of-flight (TOF) and quadrupole-time-of-flight (Q-TOF) analyzers has even enabled the detection of larger complexes with molecular weights greatly higher than 200 kDa. In this paper, we report a comparative ESI MS study on the protein quaternary structure of native hemocyanins (Hc) from crabs living in different biotopes: a shore crab (Carcinus maenas) and two deep sea crabs (Segonzacia mesatlantica and Bythograea thermydron). Hc is an extracellular blood protein, composed of several protein chains which can associate in large multimers. The goal of this study is to point out that the oligomerization state of native Hcs is biotope-dependent. Depending on the crab, ESI-MS analyses under non-denaturing conditions reveal different oligomeric forms present in equilibrium in solution. Molecular weights up to 2,235 kDa were measured for the associations of 30 subunits of the Bythograea thermydron Hc. Thanks to ESI-MS analyses, it could be concluded for the first time that the oligomerization state of native Hcs is dependent on the crab environment. The investigation of these different non-covalent self-assemblies is very important for the life history of crabs, since they are directly related with different oxygen binding abilities and thus, with their ability to colonize habitats with different oxygen contents. PMID- 12745213 TI - Mass spectrometric determination of association constants of adenylate kinase with two noncovalent inhibitors. AB - Noncovalent complexes between chicken muscle adenylate kinase and two inhibitors, P(1),P(4)-di(adenosine-5')tetraphosphate (Ap4A) and P(1),P(5)-di(adenosine-5') pentaphosphate (Ap5A), were investigated with electrospray ionization mass spectrometry under non-denaturing conditions. The nonconvalent nature and the specificity of the complexes are demonstrated with a number of control experiments. Titration experiments allowed the association constants for inhibitor binding to be determined. Problems with concentration dependent ion yields are circumvented by a data evaluation method that is insensitive to the overall ionization efficiency. The K(a) values found were 9.0 x 10(4) M(-1) (Ap4A) and 4.0 x 10(7) M(-1) (Ap5A), respectively, in very good agreement with available literature data. PMID- 12745214 TI - Dication induced stabilization of gas-phase ternary beta-cyclodextrin inclusion complexes observed by electrospray mass spectrometry. AB - Electrospray mass spectrometry (ES-MS) is an important tool for characterization of non-covalent binding in the gas phase. In this study, iron (II) has been introduced as a dication to enhance the detection of cyclodextrin (CD) plus aromatic compound complexes in ES-MS. Evidence that a novel ternary complex comprised of one beta-CD, one iron (II) and one toluene exists as an inclusion complex has been compiled via ES-MS and ES-MS/MS experiments as well as by a computational approach. This evidence strongly suggests that iron (II) serves to modify the conformation of the beta-CD ring, and that toluene inclusion is stabilized by dication interaction with the toluene pi-system and by crimping of the beta-CD ring leading to stronger van der Waals interactions with toluene. Mg(II), another dication of similar radius, showed similar behavior, while added group one cations (H(+) and Na(+)) were ineffective at producing observable ions representative of the complex. The ternary beta-CD complex with iron (II) and 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) has also been examined. ES-MS and ES MS/MS experiments suggest that it is the polar portion of 2,4,5-T (i.e., the carboxylic acid moiety) that is favored for inclusion in the beta-CD cavity, rather than the non-polar aromatic part. PMID- 12745212 TI - Diffusion measurements by electrospray mass spectrometry for studying solution phase noncovalent interactions. AB - This study describes a novel approach for monitoring noncovalent interactions in solution by electrospray mass spectrometry (ESI-MS). The technique is based on measurements of analyte diffusion in solution. Diffusion coefficients of a target macromolecule and a potential low molecular weight binding partner are determined by measuring the spread of an initially sharp boundary between two solutions of different concentration in a laminar flow tube (Taylor dispersion), as described in Rapid Commun. Mass Spectrom. 2002, 16, 1454-1462. In the absence of noncovalent interactions, the measured ESI-MS dispersion profiles are expected to show a gradual transition for the macromolecule and a steep transition for the low molecular weight compound. However, if the two analytes form a noncovalent complex in solution the dispersion profiles of the two species will be very similar, since the translational diffusion of the small compound is determined by the slow Brownian motion of the macromolecule. In contrast to conventional ESI-MS based techniques for studying noncovalent complexes, this approach does not rely on the preservation of solution-phase interactions in the gas phase. On the contrary, "harsh" conditions at the ion source are required to disrupt any potential gas- phase interactions between the two species, such that their dispersion profiles can be monitored separately. The viability of this technique is demonstrated in studies on noncovalent heme-protein interactions in myoglobin. Tight noncovalent binding is observed in solutions of pH 10, both in the absence and in the presence of 30% acetonitrile. In contrast, a significant disruption of the noncovalent interactions is seen at an acetonitrile content of 50%. Under these conditions, the diffusion coefficient of heme in the presence of myoglobin is only slightly lower than that of heme in a protein-free solution. A breakdown of the noncovalent interactions is also observed in aqueous solution of pH 2.4, where myoglobin is known to adopt an acid-unfolded conformation. PMID- 12745215 TI - Mass spectrometry and non-covalent protein-ligand complexes: confirmation of binding sites and changes in tertiary structure. AB - An experimental approach is described for determining protein-small molecule non covalent ligand binding sites and protein conformational changes induced by ligand binding. The methodology utilizes time resolved limited proteolysis and the high throughput analysis capability of MALDI TOF MS to determine the binding site in a tetanus toxin C-fragment (51 kDa)-doxorubicin (543 Da) non-covalent complex. Comparing relative ion abundances of peptides released from the time resolved limited proteolysis of tetanus toxin C-fragment (TetC) and the TetC doxorubicin complex every 10 min from 10 to 120 min of digestion revealed that the binding of doxorubicin induced a significant change in surface topology of TetC. Four of the twenty-nine peptides observed by MALDI MS, including amino acids 351-360, 299-304, 305-311 and 312-316, had a lower abundance in the TetC doxorubicin complex relative to TetC from 10 to 100 min of digestion. A decrease in ion abundance suggests doxorubicin obstructs the access of the protease to one or both termini of these peptides, identifying doxorubicin binding site(s). Conversely, five peptide ions, including amino acids 335-350, 364-375, 364-376, 281-298, and 316-328, all had a greater abundance in the digest of the complex, indicating an increase in accessibility to these sites. These five peptides flank regions of decreased ion abundance, suggesting that doxorubicin not only binds to the surface, but also induces a conformational change in TetC. PMID- 12745216 TI - Hydrogen/deuterium exchange on yeast ATPase supramolecular protein complex analyzed at high sensitivity by MALDI mass spectrometry. AB - To evaluate the ability of hydrogen/deuterium exchange of amide protons followed by mass spectrometry (HXMS) to yield topological information about supramolecular protein complexes, this approach has been tested with the 370 kDa hetero oligomeric complex of yeast F1-ATPase. The study was focused on the epsilon subunit (6612 Da) of the complex. Deuterium back exchange due to the chromatographic isolation step of this subunit was strongly reduced by means of fast micro-chromatography, and MALDI-MS was used to analyze either the intact subunit or peptide mixtures resulting from its proteolytic cleavage. A deuterium labeling kinetic study was conducted with epsilon subunit being a part of the F1 native complex. The effect of a secondary structure was also investigated by means of HXMS on the isolated epsilon subunit. Finally, to determine which regions of epsilon subunit are accessible to solvent in F1-ATPase during exchange, the complex was submitted to hydrogen/deuterium exchange, the epsilon subunit was purified by micro-chromatography, digested by pepsin, and resulting peptide fragments were analyzed by MALDI-MS. The combination of hydrogen/deuterium exchange, fast micro-chromatography and MALDI-MS was shown to be a fast and efficient way to obtain detailed topological information for the epsilon subunit when it is engaged in the ATPase complex. PMID- 12745217 TI - Detection of estrogen DNA-adducts in human breast tumor tissue and healthy tissue by combined nano LC-nano ES tandem mass spectrometry. AB - For the first time estrogen DNA-adducts were identified in DNA human breast tumor tissue using nano-LC coupled to nano-Electrospray Tandem Mass Spectrometry. Normal breast tissue was analyzed analogously. The data obtained in the five breast tumor and five adjacent normal tissue samples were compared qualitatively, but no straightforward difference was observed. Prior to LC-MS analysis the DNA was enzymatically hydrolyzed to a nucleoside pool. The DNA-hydrolysates were directly injected onto a column switching system developed for on-line sample clean-up and subsequent analysis of the DNA-adducts. In four patients using Premarin, DNA-adducts of 4-hydroxy-equilenin (4OHEN) were detected. All except three samples contained DNA-adducts from 4-hydroxy-estradiol or 4-hydroxy estrone. Also DNA isolated from eight alcohol fixed and paraffin embedded breast tumor tissue showed the presence of different estrogen DNA-adducts. Worthwhile mentioning is the presence of adducts responding to m/z 570 > m/z 454 transition. This is a well-known SRM-transition indicative for the presence of the 2' deoxyguanosine (dGuo) adduct of Benzo[a]pyrene. PMID- 12745218 TI - Effect of different solution flow rates on analyte ion signals in nano-ESI MS, or: when does ESI turn into nano-ESI? AB - In nano-ESI MS, the qualitative and quantitative characteristics of mass spectra vary considerably upon the use of different spraying conditions, i.e., aperture of the spraying needle and the voltage applied. The major parameters affected by the aperture size is the liquid flow rate which determines the initial droplet size and the current emitted upon the spray process, as described by different models of the ESI process. In the present study, the effect of flow rate on ion signals was studied systematically using mixtures of compounds with different physicochemical properties (i.e., detergent/oligosaccharide and oligosaccharide/peptide). For these model systems, the functional dependence of certain analyte-ion ratios upon the flow rate can be correlated to changes in analyte partition during droplet fission prior to ion release. Analyte suppression is practically absent at minimal flow rates below 20 nL/min. PMID- 12745219 TI - Collision rate constants for polarizable ions. AB - Langevin described a model for the interaction between an ion and a neutral nearly a century ago and since then, many modifications have been introduced to adjust for specific circumstances. This work discusses the induced dipole-induced dipole interaction between an ion and a neutral without a permanent dipole and introduces an anisotropic adjustment. A point polarizable ion model (PPI) and an orientation dependent polarizable ion model (ODPI) are discussed and applied to systems where the ion is highly polarizable and the neutral is only weakly polarizable. Significant deviations from classical Langevin rate constants and significant differences between PPI and ODPI are observed. PMID- 12745221 TI - Low nanogram per liter determination of halogenated nonylphenols, nonylphenol carboxylates, and their non-halogenated precursors in water and sludge by liquid chromatography electrospray tandem mass spectrometry. AB - A new LC-MS-MS method for quantitative analysis of nonylphenol (NP), nonylphenol carboxylates (NPECs), and their halogenated derivatives: brominated and chlorinated nonylphenols (BrNP, ClNP), brominated and chlorinated nonylphenol carboxylates (BrNPE(1)C and ClNPE(1)C) and ethoxycarboxylates (BrNPE(2)C and ClNPE(2)C) in water and sludge has been developed. Electrospray negative ionization MS-MS was applied for the identification of above mentioned compounds. Upon collision-induced dissociation, their deprotonated molecules gave different fragments formed by the cleavage of the alkyl moiety and/or (ethoxy)carboxylic moiety. For halogenated compounds a highly diagnostic characteristic pattern of isotopic doublet signals was obtained and fragmentation yielded, in addition to above mentioned ions, [Br](-) and [Cl](-), respectively. Quantitative analysis was done in the multiple reaction monitoring (MRM) mode, using two specific combinations of a precursor-product ion transitions for each compound. Additionally, for halogenated compounds two specific channels for each transition reaction, corresponding to two isotopes, were monitored and the ratio of their abundances used as an identification criterion. The method has been validated in terms of sensitivity, selectivity, accuracy, and precision and was applied to the analysis of water and sludge samples from drinking water treatment plant (DWTP) of Barcelona (Catalonia, NE Spain). Halogenated NP and NPECs were detected in prechlorinated water in concentrations up to 315 ng/L, BrNPE(2)C being the most abundant compound. In the DWTP effluent non-halogenated compounds were detected at trace levels (85, 12 and 10 ng/L for NP, NPE(1)C, and NPE(2)C, respectively), whereas concentration of halogenated derivatives never exceeded 10 ng/L. Nonylphenol, brominated and chlorinated NPs were found in flocculation sludge in concentrations of 150, 105, and 145 microg/kg, respectively. Acidic polar metabolites were found in lower concentrations up to 20 microg/kg. PMID- 12745220 TI - Indirect assessment of small hydrophobic ligand binding to a model protein using a combination of ESI MS and HDX/ESI MS. AB - Direct mass spectrometric characterization of interactions between proteins and small hydrophobic ligands often poses a serious problem due to the complex instability in the gas phase. We have developed a method that probes the efficacy of ligand-protein interactions indirectly by monitoring changes in protein flexibility. The latter is assessed quantitatively using a combination of charge state distribution analysis and amide hydrogen exchange under both native and mildly denaturing conditions. The method was used to evaluate binding of a model protein cellular retinoic acid binding protein I to its natural ligand all-trans retinoic acid (RA), isomers 13-cis- and 9-cis-RA, and retinol, yielding the following order of ligand affinities: All-trans RA > 9-cis RA > 13-cis RA, with no detectable binding of retinol. This order is in agreement with the results of earlier fluorimetric titration studies. Furthermore, binding energy of the protein to each of retinoic acid isomers was determined based on the measured hydrogen exchange kinetics data acquired under native conditions. PMID- 12745222 TI - Establishing the fitness for purpose of mass spectrometric methods. AB - This report is submitted by a working group sponsored by the ASMS Measurements and Standards Committee. The group responded to a 1998 opinion piece dealing with mass spectrometry in trace analysis (Bethem, R. A.; Boyd, R. K. J. Am. Soc. Mass Spectrom. 1998, 9, 643-648) which proposed that the concept of fitness for purpose addresses the needs of a wide range of analytical problems. There is a need to define fitness for purpose within the current context of mass spectrometry and to recommend processes for developing and evaluating methods according to suitability for a particular purpose. The key element in our proposal is for the interested parties to define in advance the acceptable degree of measurement uncertainty and the desired degree of identification confidence. These choices can serve as guideposts during method development and targets for retrospective evaluation of methods. A series of more detailed recommendations are derived from basic principles and also from reviews of current practice. This report highlights some areas where consensus is evident, but also revealed the need for further work in other areas. The recommendations are aimed primarily for the laboratory analyst but we hope they will be accessible to the non-scientist as well. Our goal was to provide a framework that can support informed decisions and foster discussion of the issues, because ultimately it is the responsibility of the analyst to make choices, provide supporting data, and interpret results according to scientific principles and qualified judgment. PMID- 12745223 TI - Mass spectrometry PittCon 2003. PMID- 12745229 TI - Bioprotectives and probiotics for dry sausages. AB - The microbial stability of dry sausages is determined by the combination and timing of different factors referred to as the hurdle-concept. However, the hurdles present in dry sausage are not sufficient to prevent the survival of Listeria monocytogenes or enterohemorrhagic Escherichia coli O157:H7. Recently bioprotective lactic acid bacteria, which in addition to the production of antimicrobial lactic acid, have been found to contribute to the safety of the dry sausage by producing antimicrobial peptide, i.e. bacteriocins and other low molecular-mass compounds. Furthermore, the possibilities to use probotics in dry sausage manufacturing process has been addressed recently. As one possible mode of action for probiotics is the production of antimicrobial compounds, lactic acid bacteria may act as both probiotic and bioprotective culture as well as fermenting agent in meat product, such as dry sausage. PMID- 12745230 TI - The bacterial flora of fresh and chill-stored pork: analysis by cloning and sequencing of 16S rRNA genes. AB - The composition of the initial bacterial flora of pork and the development of the flora after storage at +4 degrees C for 4 days were analysed by amplification, cloning and sequencing of 16S rDNA. A total of 122 clones were obtained, with lengths of > or =400 nucleotides and > or =95% similarity to database sequences. Nineteen clones were similar to sequences in database not assigned to any genera. Fourteen different genera were represented in clones from fresh meat, with 36.5% of the clones most resembling Acinetobacter and 17.3% resembling Staphylococcus and Macrococcus. After storage, the clones were composed of six different genera, with 44.3% resembling Pseudomonas, 17.1% resembling Aeromonas and only 14.3% resembling Acinetobacter. This study shows that the overall pattern of the initial and chill-stored pork flora, as shown by a molecular approach, was in agreement with results obtained in previous studies using traditional cultivation methods. PMID- 12745231 TI - Use of an electronic tongue to analyze mold growth in liquid media. AB - The feasibility of employing an electronic tongue to measure the growth of mold in a liquid medium was studied. We used the electronic tongue developed at Linkoping University, which is based on pulsed voltammetry and consists of an array of different metal electrodes. Instead of focusing on a single parameter, this device provides information about the condition or quality of a sample or process. Accordingly, the data obtained are complex, and multivariate methods such as principal component analysis (PCA) or projection to latent structures (PLS) are required to extract relevant information. A gas chromatographic technique was developed to measure ergosterol content in mold biomass and was subsequently used as a reference method to investigate the ability of the electronic tongue to measure the growth of mold in liquid media. The result shows that the electronic tongue can monitor mold growth in liquids. In PLS analysis, the electronic tongue signals correlate well with the amount of ergosterol in the mold biomass as well as the microbially induced changes in the pH of the medium. PMID- 12745232 TI - Relation between microbiological quality, metabolite production and sensory quality of equilibrium modified atmosphere packaged fresh-cut produce. AB - The quality of four types of fresh-cut produce, packaged in consumer-sized packages under an equilibrium modified atmosphere and stored at 7 degrees C, was assessed by establishing the relation between the microbial outgrowth and the corresponding production of nonvolatile compounds and related sensory disorders. In vitro experiments, performed on a lettuce-juice-agar, demonstrated the production of nonvolatile compounds by spoilage causing lactic acid bacteria and Enterobacteriaceae. Pseudomonas fluorescens and yeasts, however, were not able to produce detectable amounts of nonvolatile metabolites. The type of spoilage and quality deterioration in vivo depended on the type of vegetable. Mixed lettuce and chicory endives, leafy tissues, containing naturally low concentrations of sugars, showed a spoilage dominated by Gram-negative microorganisms, which are not producing nonvolatile compounds. Sensory problems were associated with visual properties and the metabolic activity of the plant tissue. Mixed bell peppers and grated celeriac, on the other hand, demonstrated a fast and intense growth of spoilage microorganisms, dominated by lactic acid bacteria and yeasts. This proliferation resulted in detectable levels of organic acids and the rejection by the trained sensory panel was based on the negative perception of the organoleptical properties (off-flavour, odour and taste). The applied microbiological criteria corresponded well with detectable changes in sensory properties and measurable concentrations of nonvolatile compounds, surely in the cases where lactic acid bacteria and yeasts were provoking spoilage. Consequently, the freshness of minimally processed vegetables, sensitive for outgrowth of lactic acid bacteria and yeasts (e.g., carrots, celeriac, bell peppers, mixtures with non-leafy vegetables) can be evaluated via analysis of the produced nonvolatile compounds. PMID- 12745233 TI - The effect of abrupt osmotic shifts on the lag phase duration of foodborne bacteria. AB - The effects of osmotic environment and inoculum history on lag times were examined. Abrupt osmotic shifts of cultures were found to induce lag phases in a variety of foodborne bacteria. Relative lag times (RLT; the ratio of lag time to generation time) were used to differentiate the effects of the shift from those of the outgrowth environment. In general, osmotic downshifts induced larger RLTs than equivalent upshifts. An observed reduction in RLT at very low a(w), however, was unexpected. For an osmotic downshift, differences were observed in the RLT response of the Gram-negative and -positive strains tested. RLTs were usually extended for Gram-negative organisms as conditions became less favourable for growth. In comparison, RLT remained relatively unaffected for Gram-positive organisms. The observations reported in this study demonstrate that lag time can be understood in terms of the amount of work to be done to adjust to new environmental conditions and the rate at which that work is done, and are consistent with known strategies for osmoregulation employed by the various organisms studied. PMID- 12745234 TI - The effect of abrupt shifts in temperature on the lag phase duration of Escherichia coli and Klebsiella oxytoca. AB - The effect of temperature of incubation on lag times of two gram-negative foodborne bacteria was investigated. Bacteria were instantaneously transferred between temperatures within and beyond the normal physiological temperature range (NPTR). Abrupt temperature shifts induced lag phases, but the degree of the response was dependent on the direction and magnitude of the shift. Temperature downshifts induced larger relative lag times (RLT; the ratio of lag time to generation time), than equivalent upshifts. The hypothesis of Robinson et al. [Int. J. Food Microbiol. 44 (1998) 83] that lag time can be understood in terms of the amount of work to be done to adjust to new environmental conditions and the rate at which that work is done was supported. Deviation of the reported proportionality between lag time and generation time was observed when late exponential phase cells were subjected to abrupt temperature shifts from beyond the normal physiological range. PMID- 12745235 TI - Use of starter cultures of lactic acid bacteria and yeasts in the preparation of togwa, a Tanzanian fermented food. AB - Starter cultures of lactic acid bacteria (Lactobacillus brevis, Lactobacillus cellobiosus, Lactobacillus fermentum, Lactobacillus plantarum and Pediococcus pentosaceus) and yeasts (Candida pelliculosa, Candida tropicalis, Issatchenkia orientalis and Saccharomyes cerevisiae) isolated from native togwa were tested singly or in combination for their ability to ferment maize-sorghum gruel to produce togwa. All species of bacteria showed an ability to ferment the gruel as judged by lowering the pH from 5.87 to 3.24-3.49 and increasing the titratable acidity from 0.08% to 0.30-0.44% (w/w, lactic acid) in 24 h. Yeasts used singly showed little activity within 12 h, but lowered the pH to 3.57-4.81 and increased the acidity to 0.11-0.21% in 24 h. Yeasts in co-culture with lactic acid bacteria (LAB) had a modest effect on the final acidity (P<0.05). The number of lactic acid bacteria and yeasts increased while the Enterobacteriaceae decreased with fermentation time. The pH was lowered and lactic acid produced significantly (P<0.05) fastest in natural togwa fermentation and in samples fermented by L. plantarum or L. plantarum in co-culture with I. orientalis. The content of fermentable sugars was reduced during fermentation. Most volatile flavour compounds were produced in samples from fermentation by P. pentosaceus and I. orientalis in co-culture with either L. plantarum or L. brevis. PMID- 12745236 TI - Potential use of antioxidants for control of growth and fumonisin production by Fusarium verticillioides and Fusarium proliferatum on whole maize grain. AB - The effect of interactions between two food grade antioxidants butylated hydroxyanisole (BHA) and propyl paraben (PP, 100, 200, 500 microg g(-1)) and water activity (a(w), 0.995, 0.98, 0.95) of irradiated maize on lag phase prior to growth, growth rate and fumonisin production by Fusarium verticillioides and Fusarium proliferatum was evaluated at 25 degrees C. Both antioxidants had an effect on growth characteristics, and fumonisin production. However, this was dependent on the dose used and the a(w) treatment. At 500 microg g(-1) BHA and PP increased the lag phase prior to growth, and reduced the growth rate of both Fusarium species significantly, especially at 0.95 a(w). Both antioxidants significantly reduced the production of fumonisin by both Fusarium species, especially at 0.98 and 0.95 a(w). These results suggest that these antioxidants have potential for treatment of maize grain for controlling growth of these mycotoxigenic species and prevent fumonisin accumulation. PMID- 12745237 TI - Susceptibility of Listeria monocytogenes isolated from food in Italy to antibiotics. AB - The susceptibility of 148 strains of Listeria monocytogenes isolated from food to antibiotics currently used in veterinary and human therapy was determined by standard agar dilution and disk diffusion methods. The antibiotics included amikacin, amoxicillin, cefazolin, chloramphenicol, erythromycin, flumequine, fosfomycin, gentamicin, kanamycin, lincomycin, oxytetracycline, rifampicin, spiramycin, streptomycin, tetracycline, tobramycin and vancomycin. Soussy's breakpoints and MIC(50)-MIC(90) values were used to classify the strains into sensitive, moderately sensitive and resistant groups. This work is part of a wider surveillance program on listeriosis started in Italy in 1995. PMID- 12745238 TI - Antibacterial activity of Capsicum extract against Salmonella typhimurium and Pseudomonas aeruginosa inoculated in raw beef meat. AB - The inhibitory effect of the extract of Capsicum annuum bell pepper type was evaluated against Salmonella typhimurium and Pseudomonas aeruginosa, inoculated in minced beef meat mixed with different concentrations of the extract, and stored at 7 degrees C for 7 days. The combined effect of C. annuum extract and sodium chloride on the bacterial growth was evaluated. The minimum inhibitory concentration of the extract to prevent the growth of S. typhimurium in minced beef was 1.5 ml/100 g of meat; the addition of 1%, 2%, 3% and 4% w/w of sodium chloride did not have any additional inhibitory effect on Salmonella. In the case of P. aeruginosa, a concentration of 0.3 ml of the extract/100 g of meat showed a bacteriostatic effect, while a concentration of 3 ml/100 g of meat showed a bactericidal effect. When 1% w/w of sodium chloride was added to the meat together with the extract, the concentration needed to kill P. aeruginosa was reduced. PMID- 12745240 TI - Cubic phase gel as a drug delivery system for topical application of 5-ALA, its ester derivatives and m-THPC in photodynamic therapy (PDT). AB - The ability of the cubic liquid-crystalline phase to incorporate and control the release of drugs of varying size and polar characteristics makes it an interesting candidate as a drug delivery system. In the present study we investigated a new potential application of the cubic phase (monoolein/water; 70:30, w/w) to deliver pro-drugs and a photosensitizer for topical application in photodynamic therapy (PDT). Therefore the pro-drug 5-aminolevulinic acid (5-ALA, a PpIX precursor), its ester derivatives (hexylester, octylester and decylester), and the chlorine compound meso-tetra(hydroxyphenyl)chlorine (m-THPC) were incorporated into the cubic phase gel of monoolein/water and their physicochemical and spectroscopic properties were investigated at 37 degrees C. Drug stability was monitored for short and long periods of time. 5-ALA and its ester derivatives as non-fluorescent probes had their properties studied after chemical reaction leading to a fluorescent derivative. For all the compounds analyzed in this study the spectroscopic properties were clearly defined with potential photodynamic activity in the gel formulation. We are currently evaluating the potential of monoolein/water as a drug delivery system in the treatment of different cutaneous diseases and other PDT applications. PMID- 12745241 TI - The plasma membrane-associated NADH oxidase (ECTO-NOX) of mouse skin responds to blue light. AB - NADH oxidases of the external plasma membrane surface (ECTO-NOX proteins) are characterized by oscillations in activity with a regular period length of 24 min. Explants of mouse skin exhibit the oscillatory activity as estimated from the decrease in A(340) suggesting that individual ECTO-NOX molecules must somehow be induced to function synchronously. Transfer of explants of mouse skin from darkness to blue light (495 nm, 2 min, 50 micromol m(-1) s(-1)) resulted in initiation of a new activity maximum (entrainment) with a midpoint 36 min after light exposure followed by maxima every 24 min thereafter. Addition of melatonin resulted in a new maximum 24 min after melatonin addition. The findings suggest that the ECTO-NOX proteins play a central role in the entrainment of the biological clock both by light and by melatonin. PMID- 12745242 TI - Endoscopic light-induced autofluorescence spectroscopy for the diagnosis of colorectal cancer and adenoma. AB - To evaluate the new, bio-optical method of light-induced autofluorescence spectroscopy for the endoscopic in-vivo diagnosis of (pre)-cancerous lesions of the colorectum, 311 endogenous fluorescence spectra were obtained from normal, adenomatous and cancerous colorectal tissue in 11 patients with cancer, six patients with familial adenomatous polyposis, and six patients with multiple adenomatous polyps. A light source delivered either white or violet-blue light for excitation of tissue autofluorescence via a flexible endoscope. Endogenous fluorescence spectra emitted by the tissue were picked up with a fiberoptic probe and analysed with a spectrograph. Biopsies were taken for definitive classification of the spectra. Rectal cancer (n=11) as well as adenomas with severe dysplasia (n=19) showed specific differences between the emitted fluorescence spectra as compared with normal mucosa and hyperplastic polyps. Having applied a mathematical algorithm to the spectra, a sensitivity of 96% and a specificity of 93% were obtained for the diagnosis of rectal cancer. The equivalent values for the diagnosis of dysplastic ademomas were 98 and 89%, respectively. Light-induced autofluorescence spectroscopy is a new and promising bio-optical procedure for the endoscopic in-vivo diagnosis of colorectal cancer and dysplasia. PMID- 12745243 TI - Spectral analysis of pigment photobleaching in photosynthetic antenna complex LHCIIb. AB - Light-induced photooxidation of chlorophyll (Chl) a, b and xanthophylls was investigated in LHCIIb, the antenna pigment-protein complex of photosystem II. Absorption difference spectra at normal and low temperatures show initially (at less than 25% Chl a decay) a selective bleaching of a red-shifted Chl b with absorption bands at 487 and 655 nm, Chl b (460/650 nm) and Chl a (433/670 nm), which changes to a less selective photooxidation pattern at deeper bleaching stages. Difference absorption spectra and HPLC analyses indicate different photooxidation rates of pigments in the order neoxanthin>Chl a>lutein approximately Chl b. Despite significant pigment loss as monitored with absorption spectra, CD spectra indicate an essentially complete persistence of the protein secondary structure. Fluorescence excitation spectra suggest the conversion of a small fraction of Chl a into pheophytin a which acts as a fluorescence quencher, possibly through temporary charge separation process. The strong features in the electroabsorption (Stark effect) spectra due to chlorophyll b at 655 nm and a xanthophyll at 510 nm, and the spectral changes mentioned above are assigned to Chl molecules located at several binding sites in LHCIIb protein and are discussed in the context of spatial configuration and interactions of pigment molecules. PMID- 12745244 TI - Photoprotective properties of a hydrophilic extract of the fern Polypodium leucotomos on human skin cells. AB - The effect of a hydrophilic extract of the fern Polypodium leucotomos (PLE) has been investigated in terms of photoprotection against UV-induced cell damage. PLE efficiently preserved human fibroblast survival and restored their proliferative capability when the cells were exposed to UVA light. This effect was specific and dose-dependent. Photoprotection was not restricted to fibroblasts, as demonstrated by its effect on survival and proliferation of the human keratinocyte cell line HaCat. Finally, treatment of the cells with PLE prevented UV-induced morphological changes in human fibroblasts, namely disorganisation of F-actin-based cytoskeletal structures, coalescence of the tubulin cytoskeleton and mislocalization of adhesion molecules such as cadherins and integrins. Our in vitro results demonstrate the photoprotective effect of PLE on human cells and support its use in the preventive treatment of sunburning and skin pathologies associated with UV-mediated damage. PMID- 12745245 TI - The effect of 880 nm low level laser energy on human fibroblast cell numbers: a possible role in hypertrophic wound healing. AB - Low level lasers (LLLs) have been shown to induce therapeutic effects in wound healing. However, there have been few LLL studies on burn wounds which may become unsightly, hypertrophic and impair function. Inhibitory effects on the healing of fibrotic wounds, prone to hypertrophy may be expected to reasonably reduce the problems accompanying hypertrophic scarring. The effects of LLL wavelengths and treatment parameters on wound healing cells in vitro often demonstrate meaningful results and without concurrent ethical difficulties of clinical trials. This experiment investigated the effect of an 880 nm, 16 mW GaAlAs diode at 2.4 and 4 J/cm(2) on cell numbers of two human fibroblast cell lines, derived from hypertrophic scar (HF) and normal dermal explants (NF), respectively. After irradiation by 880 nm LLL, cell numbers were measured utilising methylene blue bioassay and read by the spectrophotometer in the same microculture plates. HF and NF exhibited decreased cell numbers as compared to sham-irradiated controls. HF cell number, after 2.4 J/cm(2), was significantly lower on day 5 (P<0.05). The NF cell numbers were significantly lower on day 4 and/or day 5 (P<0.05). The results have implications on hypertrophic wound healing and further studies are required. PMID- 12745246 TI - TEM evidence of ultrastructural alteration on Pseudomonas aeruginosa by photocatalytic TiO2 thin films. AB - The antibacterial efficiency of longwave UV-irradiated TiO(2) thin films as well as the ultrastructural damage on bacterial cells was evaluated using Pseudomonas aeruginosa as a model. The quantitative antibacterial efficiency assays showed a bacterial inhibition in the range of 32-72% at different times of irradiation. Transmission electron microscopy (TEM) was used to detect the effect of irradiation of TiO(2) thin films on the ultrastructure of the bacterial cell in order to reveal possible cellular damage. After 40 min irradiation, an abnormal cellular division was observed: instead of a normal septum, an 'elongated bridge' was formed. At a longer irradiation time, wavy structures all around the outer cell membrane were observed, and also some bubble-like protuberances, which expelled inner material. The mechanism of irreversible bacterial cell damage caused by the photocatalytic effect of TiO(2) could be related to abnormal cell division, aside from the reported physicochemical alteration of the cell membrane. PMID- 12745247 TI - Photoactive extracts from Thevetia peruviana with antifungal properties against Cladosporium cucumerinum. AB - Seeds of Thevetia peruviana were screened for their antifungal photoactivity. Extracts obtained either with n-hexane or dichloromethane were fractionated by column chromatography and further analysed by thin-layer chromatography. All seed extracts and fractions were tested for inhibition of the fungus Cladosporium cucumerinum for the evaluation of photoactive inhibitory effects. Antifungal light-dependent activity was observed for some of the fractions and both crude extracts. The most photoactive fraction was analysed by capillary gas chromatography with mass spectrometry in order to identify its constituents. Two major groups of compounds were identified, terpenes and fatty acids and derivatives. Pulegone, linoleic acid and palmitic acid were the major compounds. Terpenes seem to be the major substances with antifungal photoactivity. PMID- 12745248 TI - Diastereoisomerism, contact points, and chiral selectivity: a four-site saga. AB - In biology, chiral recognition usually implies the ability of a protein, such as an enzyme or a drug receptor, to distinguish between the two enantiomeric forms of a chiral substrate or drug. Both diastereoisomerism and specific contacts between enzyme/receptor and substrate/drug are necessary. The minimum requirement is for four contact points including four nonplanar atoms (or groups of atoms) in both probe and target. The molecular models described by Easson and Stedman and by Ogston require three binding sites in a plane. A modified model with three binding sites in three dimensions is described. Under certain circumstances this model allows binding of both enantiomeric forms of a substrate or a drug. Enantiomer superposition of two enantiomers at an active site occurs in some specific cases (e.g., phenylalanine ammonia-lyase, isocitrate dehydrogenase) and is likely in others. The nature of enantiomer binding to racemase enzymes is discussed. PMID- 12745249 TI - Gene-targeting reveals physiological roles and complex regulation of the phosphoinositide 3-kinases. AB - Phosphoinositide 3-kinases (PI3Ks) are represented by a family of eight distinct enzymes that can be divided into three classes based on their structure and function. The class I PI3Ks are heterodimeric enzymes that are regulated by recruitment to plasma membrane following receptor activation and which control numerous cellular functions, including growth, differentiation, migration, survival, and metabolism. New light has been shed on the biological role of individual members of the class I PI3Ks and their regulatory subunits through gene-targeting experiments. In addition, these experiments have brought the complexity of how PI3K activation is regulated into focus. PMID- 12745250 TI - Endogenous oxidoreductase expression is induced by aminoglycosides. AB - Oxidoreductases such as glutaredoxin are a major class of enzymes that reversibly catalyze thiol-disulfide exchange reactions. Transfection experiments using geneticin (G418) selection to identify the specific protein S-thiolated substrates of glutaredoxin-1 (Grx-1) noted the curious phenomenon that nontransfected control cells treated with G418 had increased levels of Grx-1 expression. Varied concentrations of gentamicin, kanamycin, and hygromycin increased Grx-1 expression in a time- and dose-dependent fashion in human cultured retinal pigment epithelial cells. Reactive oxygen species formation after aminoglycoside exposure correlated directly to aminoglycoside treatment. Further indication that oxidation regulates Grx-1 expression was noted by the positive effect of phorbol 12-myristate 13-acetate, a known inducer of redox sensitive AP-1 transcription factor. In agreement with this hypothesis was the finding that the physiologic reductant N-acetylcysteine decreased Grx-1 expression whereas tert-butyl hydroperoxide increased Grx-1 expression. Our data suggest that aminoglycosides increased Grx-1 expression in response to oxidative stress. PMID- 12745251 TI - A SWI/SNF-like factor from chicken liver that disrupts nucleosomes and transfers histone octamers in cis and trans. AB - ATP-dependent chromatin remodeling factors have been implicated in nuclear processes involving DNA. Here we report partial purification and characterization of an ATP-dependent chromatin remodeling activity from chicken liver. Nuclear extract from chicken liver was fractionated chromatographically to enrich proteins immunoreacting to antibodies against components of human SWI/SNF, namely BRG1, BAF170, BAF155, and BAF57. Immunoreactivity to these antibodies elutes with a mass of about 2MDa on Sepharose CL-6B gel filtration, suggesting that they constitute a SWI/SNF-like complex (SLC). The SLC displays three chromatin remodeling activities, viz. nucleosome disruption, octamer transfer, and nucleosome sliding (octamer transfer in cis). We further show that components of SLC, as revealed by immunoreactivity to the above antibodies, display a dynamic nucleocytoplasmic distribution and colocalize with RNA polymerase II in the liver nuclei. This report contributes to the understanding of phylogenetic generality of chromatin remodeling factors in eukaryotes. PMID- 12745252 TI - Adenosyl coenzyme and pH dependence of the [4Fe-4S]2+/1+ transition in lysine 2,3 aminomutase. AB - 5'-[N-[(3S)-3-Amino-carboxypropyl]-N-methylamino]-5(')-deoxyadenosine (azaSAM), an analog of S-adenosyl-L-methionine (SAM), was used to study the cofactor dependent reduction of the [4Fe-4S](2+) center in lysine 2,3-aminomutase to the +1 oxidation state. azaSAM has a tertiary nitrogen in place of the sulfonium center of SAM. The analog binds to lysine 2,3-aminomutase with K(d)s of 1.4+/-0.3 microM at pH 8.0 and 2.2+/-0.6 microM at pH 6.5. Reduction of the [4Fe-4S](2+) center in the presence of this analog gives a 10K [4Fe-4S](1+) electron paramagnetic resonance (EPR) signal similar to that seen with SAM or S-adenosyl-L homocysteine (SAH). The pH dependence of cofactor-induced reduction was examined to determine whether ionization of the tertiary nitrogen (pK(a)=7.08) might affect reduction of the [4Fe-4S](2+) center. The results show similar behavior in azaSAM and SAH, demonstrating that ionization of the aza group in azaSAM does not account for pH dependence in cofactor-dependent reduction of the [4Fe-4S](2+) center. The signal shape of the low-temperature EPR signal for the [4Fe-4S](1+) center in the SAM-induced reduction displayed a pH dependence that was not observed in the azaSAM- or SAH-induced spectra. Unique features of the signal are at a maximum at the pH activity optimum of pH 8 and are diminished as the pH is lowered or raised. These features are also absent in the spectra at all pHs examined when reduction is induced by azaSAM or SAH. PMID- 12745253 TI - Purification and characterization of T cell protein tyrosine phosphatase reveals significant functional homology to protein tyrosine phosphatase-1B. AB - We have developed a protocol for rapid purification of T cell protein tyrosine phosphatase (TCPTP) and the structurally related protein tyrosine phosphatase-1B (PTP-1B) from bacterial cells. The pH profile for TCPTP was bell-shaped with an optimum of 5.5. The catalytic domain and full-length versions of TCPTP bound a potent inhibitor with affinities similar to those of PTP-1B. The K(m) values for the catalytic domains of TCPTP and PTP-1B increased with increasing ionic strength, whereas the k(cat) values remained unchanged. Arrhenius plots revealed that TCPTP and PTP-1B possess similar activation energies of 25.3+/-1.2 and 18.4+/-3.0 kJ/mol, respectively. Increasing solvent microviscosity (up to 40% (w/v) sucrose) did not affect k(cat)/K(m) of either enzyme. However, high sucrose concentrations protected both enzymes from thermal inactivation. These studies show that, although they share a 72% amino acid sequence identity within their catalytic domains, TCPTP and PTP-1B are functionally very similar in vitro. PMID- 12745254 TI - Heterologous expression, purification, and enzymatic characterization of the acyclic carotenoid 1,2-hydratase from Rubrivivax gelatinosus. AB - The carotenoid 1,2-hydratase CrtC from Rubrivivax gelatinosus has been expressed in Escherichia coli in an active form and purified by affinity chromatography. The enzyme catalyzes the conversion of various acyclic carotenes including 1 hydroxy derivatives. This broad substrate specificity reflects the participation of CrtC in 1'-HO-spheroidene and in spirilloxanthin biosynthesis. Enzyme kinetic studies including the determination of substrate specificity constants indicate that among the alternative biosynthetic routes to 1'-HO-spheroidene the one via spheroidene is the dominating pathway. In contrast to CrtC from Rvi. gelatinosus, the equivalent enzyme from Rhodobacter capsulatus, a closely related bacterium which lacks the biosynthetic branch to spirilloxanthin and accumulates spheroidene instead of substantial amounts of 1'-HO-spheroidene, is extremely poor in converting 1-HO-carotenoids. The individual catalytic properties of both carotenoid 1,2-hydratases reflect the in situ carotenogenic pathways in both purple photosynthetic bacteria. PMID- 12745255 TI - Aging defect at the QO site of complex III augments oxyradical production in rat heart interfibrillar mitochondria. AB - Complex III in the mitochondrial electron transport chain is a proposed site for the enhanced production of reactive oxygen species that contribute to aging in the heart. We describe a defect in the ubiquinol binding site (Q(O)) within cytochrome b in complex III only in the interfibrillar population of cardiac mitochondria during aging. The defect is manifested as a leak of electrons through myxothiazol blockade to reduce cytochrome b and is observed whether cytochrome b in complex III is reduced from the forward or the reverse direction. The aging defect increases the production of reactive oxygen species from the Q(O) site of complex III in interfibrillar mitochondria. A greater leak of electrons from complex III during the oxidation of ubiquinol is a likely mechanism for the enhanced oxidant production from mitochondria that contributes to aging in the rat heart. PMID- 12745256 TI - cDNA cloning, expression, and functional characterization of a zebrafish SULT1 cytosolic sulfotransferase. AB - Using the reverse transcriptase-polymerase chain reaction technique, a full length cDNA encoding a novel zebrafish sulfotransferase was cloned and sequenced. Sequence analysis indicated that this zebrafish sulfotransferase belongs to the SULT1 cytosolic sulfotransferase gene family. The recombinant form of the zebrafish sulfotransferase, purified from Escherichia coli cells, displayed sulfating activities toward a number of endogenous compounds, in particular dopamine and thyroid hormones, in addition to xenobiotics including some flavonoids, isoflavonoids, and other phenolic compounds. The zebrafish sulfotransferase exhibited substrate dependence in pH optimum. In comparison with those determined with dopamine as substrate, the zebrafish sulfotransferase displayed much lower K(m) and higher V(max) with n-propyl gallate as substrate. A thermostability experiment revealed the enzyme to be relatively stable over a temperature range between 20 and 43 degrees C. Among 10 divalent metal cations tested, Hg(2+), Co(2+), Zn(2+), Cd(2+), Cu(2+), and Pb(2+) exhibited dramatic inhibitory effects on the activity of the zebrafish sulfotransferase. PMID- 12745257 TI - Rapid flip-flop in polyunsaturated (docosahexaenoate) phospholipid membranes. AB - The transbilayer movement (flip-flop) of 7-nitrobenz-2-oxa-1,3-diazol-4-yl phosphatidylethanolamine (NBD-PE) in phosphatidylcholine (PC) membranes containing various acyl chains was measured by dithionite quenching of NBD fluorescence. Of specific interest was docosahexaenoic acid (DHA), the longest and most unsaturated acyl chain commonly found in membranes. This molecule represents the extreme example of a family of important fatty acids known as omega-3s and has been clearly demonstrated to alter membrane structure and function. One important property that has yet to be reported is the effect of DHA on membrane phospholipid flip-flop. This study demonstrates that as the number of double bonds in the fatty acyl chains comprising the membrane increases, so does the rate of flip-flop of the NBD-PE probe. The increase is particularly marked in the presence of DHA. Half-lives t(1/2) of 0.29 and 0.086 h describe the process in 1-stearoyl-2-docosahexaenoylphosphatidylcholine and 1,2 didocosahexaenoylphosphatidylcholine, respectively, whereas in 1-stearoyl-2 oleoylphosphatidylcholine t(1/2)=11.5h. Enhanced permeability to dithionite with increasing unsaturation was also indicated by our results. We conclude that PC membranes containing DHA support faster flip-flop and permeability rates than those measured for other less-unsaturated PCs. PMID- 12745258 TI - Mechanism of binding of warfarin enantiomers to recombinant domains of human albumin. AB - Domain fragments of human serum albumin corresponding to domains 1 and 2 (D12) and domains 2 and 3 (D23) were expressed in yeast. The kinetics of warfarin binding to these fragments were investigated using stopped-flow fluorescence spectroscopy. Binding can be characterized by a two-step process, a rapid diffusion-controlled step and a slower rate-limiting step in which a stable drug protein complex is formed. The equilibrium constant for step 1 is greater for both D12 and D23 than for albumin, probably as a result of reduced steric hindrance offered by the domain fragments. Binding step 2, thought to be the result of a conformational change as warfarin is accommodated by the protein, is faster for D12 and D23. Albumin and the domain fragments show an increased preference for the R enantiomer, but the preference is particularly enhanced for domain fragment D12. These preferences can largely be explained by the domains having different rates for step 2 of the binding process. PMID- 12745259 TI - Comparative expression profiling of 40 mouse cytochrome P450 genes in embryonic and adult tissues. AB - This study is the first systematic investigation of the gestational age-dependent and adult tissue-specific expression patterns of each known mouse CYP family (40 genes) using normalized cDNA panels and uniform reverse transcriptase polymerase chain reaction-based assays. Twenty-seven of the P450s were constitutively expressed during development. The number gradually increased through the phases of gastrulation E7 (n=14), neural patterning and somitogenesis E11 (n=17), organogenesis E15 (n=20), and fetal period E17 (n=21). Cyp2s1, Cyp8a1, Cyp20, Cyp21a1, Cyp26a1, Cyp46, and Cyp51 were detected in each of the four stages studied. Members of family CYP1 demonstrated complex, nonoverlapping embryonic patterns of expression, indicating that Cyp1a1 and Cyp1a2 may not compensate for Cyp1b1 deficiency associated with abnormal eye development. Multiple Cyp forms were found to be constitutively expressed in each of the adult tissues studied: liver (n=31), kidney (n=30), testis (n=26), lung (n=24), and heart (n=13). The tissue-specific P450-expression profiles reported in this study provide a reference for more focused analysis of the tissue-specific and developmental functions of the cytochrome P450 monooxygenases. PMID- 12745260 TI - Fluorescence study of ligand binding to potato tuber pyrophosphate-dependent phosphofructokinase: evidence for competitive binding between fructose-1,6 bisphosphate and fructose-2,6-bisphosphate. AB - The intrinsic fluorescence of potato tuber pyrophosphate:fructose-6-phosphate 1 phosphotransferase (PFP) was used as an indicator of conformational changes due to ligand binding. Binding of the substrates and the allosteric activator fructose-2,6-bisphosphate was quantitatively compared to their respective kinetic effects on enzymatic activity. PFP exhibited a relatively high affinity for its isolated substrates, relative to the enzyme's respective K(m) (substrate) values. There are two distinct types of fructose-1,6-bisphosphate interaction with PFP, corresponding to catalytic and activatory binding. Activatory fructose-1,6 bisphosphate binding shares several characteristics with fructose-2,6 bisphosphate binding, indicating that both ligands compete for the same allosteric activator site. Activation by fructose-1,6-bisphosphate or fructose 2,6-bisphosphate was exerted primarily on the forward (glycolytic) reaction by greatly increasing the enzyme's affinity for fructose-6-phosphate. Binding of substrates and effectors to PFP and PFP kinetic properties were markedly influenced by assay pH. Results indicate an increased glycolytic role for PFP during cytosolic acidification that accompanies anoxia stress. PMID- 12745261 TI - Replication fork-stimulated eIF-4A from Plasmodium cynomolgi unwinds DNA in the 3' to 5' direction and is inhibited by DNA-interacting compounds. AB - Plasmodium cynomolgi DEAD-box DNA helicase 45 (PcDDH45) is an ATP-dependent DNA unwinding enzyme with intrinsic DNA-dependent ATPase activity and is highly homologous to eIF-4A. In this study, we have further characterized and tested the effect of various DNA-interacting compounds on the DNA-unwinding activity of PcDDH45. The results show that PcDDH45 translocates in the 3' to 5' direction along the bound strand, a replication fork-like structure of the substrate stimulates its DNA-unwinding activity, and it failed to unwind blunt-ended duplex DNA. Of various compounds tested, only cisplatin, 4',6'-diamidino-2-phenylindole, daunorubicin, and nogalamycin were inhibitory to the unwinding activity of PcDDH45 with apparent IC(50) values of 1.0, 4.0, 7.5, and 1.7 microM, respectively. These results suggest that the interaction of these compounds with duplex DNA generate a complex that probably impedes the translocation of PcDDH45, resulting in inhibition of unwinding activity. This study is one of the first to demonstrate the effect of various DNA-binding compounds on a malaria parasite DNA helicase and should make an important contribution to our better understanding of the nucleic acid transactions in the parasite. PMID- 12745262 TI - Radical chemistry of epigallocatechin gallate and its relevance to protein damage. AB - The radical chemistry of the plant polyphenolics epigallocatechin gallate (EGCG) and epigallocatechin (EGC) were investigated using electron paramagnetic resonance spectroscopy. Radical species formed spontaneously in aqueous solutions at low pH without external oxidant and were spin stabilized with Zn(II). The spectra were assigned to the gallyl radical and the anion gallyl radical, with only 10% of the signal assigned to a radical from the galloyl ester. Spectral simulations were used to establish a pK(a) of 4.8 for the EGCG radical and a pK(a) of 4.4 for the EGC radical. The electrochemical redox potentials of EGCG and EGC varied from 1000 mV at pH 3 to 400 mV at pH 8. The polyphenolics did not produce hydroxyl radicals unless reduced metal ions such as iron(II) were added to the system. Zinc(II)-stabilized EGCG radicals were more effective protein precipitating agents than unoxidized EGCG and produced irreversibly complexed protein. EGCG and other naturally occurring polyphenolics are effective radical scavengers but their radical products have the potential to damage biological molecules such as proteins. PMID- 12745263 TI - Thermodynamic analysis of the contributions of the copper ion and the disulfide bridge to azurin stability: synergism among multiple depletions. AB - The stabilizing potential of the copper ion and the disulfide bridge in azurin has been explored with the aim of inspecting the ways in which these two factors influence one another. Specifically, whether copper and disulfide contributions to protein stability are additive has been examined. To this aim, the thermal unfolding of a copper-depleted mutant lacking the disulfide bridge between Cys3 and Cys26 (apo C3A/C26A azurin) was studied by differential scanning calorimetry. A comparison of the unfolding parameters of holo and apo C3A/C26A azurin with the apo C3A/C26A protein has shown that the effects of simultaneous copper and disulfide depletion are additive only at two temperatures: T=15 degrees C and T=67 degrees C. Within this range the presence of the copper ion and the disulfide bridge has a positive synergistic effect on azurin stability. These findings might have implications for the rational use of the stabilizing potential of copper and disulfides in copper protein engineering. PMID- 12745264 TI - Treatment of cardiac iron overload in thalassemia major. PMID- 12745265 TI - Rituximab: a new therapeutic tool for primary immune thrombocytopenic purpura? PMID- 12745266 TI - Non-myeloablative conditioning before allogeneic stem cell transplantation in adult acute lymphoblastic leukemia. PMID- 12745267 TI - Economy-class syndrome: media hype or real risk? PMID- 12745268 TI - Comparative effects of deferiprone and deferoxamine on survival and cardiac disease in patients with thalassemia major: a retrospective analysis. AB - BACKGROUND AND OBJECTIVES: Iron-induced cardiac disease remains the main cause of death in patients with thalassemia major, despite chelation therapy with deferoxamine. Deferiprone is an iron chelator that has the potential to be more effective than deferoxamine in removing intracellular iron from the heart. However, to date, no study has been designed to examine the frequency of cardiac complications and survival as the primary outcomes of a comparative study between these two chelators. This retrospective study assessed the survival and the occurrence of cardiac disease in all patients with thalassemia major treated for at least 4 years with deferiprone or deferoxamine at a single center. DESIGN AND METHODS: The patients were, on average, 18.4 years old at the start of the review period and were followed up, on average, for 6 years. At baseline there was no significant difference in the percentage of patients with cardiac disease in the two therapy groups. RESULTS: At the end of the study, cardiac dysfunction, expressed as worsening of pre-existing cardiac abnormality or development of new cardiac disease, was diagnosed in 2 (4%) of the 54 deferiprone-treated patients and in 15 (20%) of the 75 deferoxamine-treated patients, from the first to the last measurement (p = 0.007). The Kaplan Meier analysis of cardiac disease-free survival over the 5-year period was significantly more favorable in the deferiprone group (p = 0.003). INTERPRETATION AND CONCLUSIONS: None of the patients treated with deferiprone died, while 3 of the patients treated with deferoxamine died because of irreversible worsening of their cardiac condition during the study period. Findings from this study suggest that long-term therapy with deferiprone provides a greater cardio-protective effect against the toxicity of iron overload than does subcutaneous deferoxamine. Formal prospective studies are warranted to confirm this effect. PMID- 12745269 TI - Differences between the CD34+ and CD34- blast compartments in apoptosis resistance in acute myeloid leukemia. AB - BACKGROUND AND OBJECTIVES: Altered expression of members of the Bcl-2 family might account for the observed apoptosis resistance to chemotherapy in acute myeloid leukemia (AML). Given the poor prognosis associated with CD34+ expression in AML, we studied the role of spontaneous apoptosis and apoptosis regulatory proteins in sorted CD34+ and CD34- primary AML fractions. DESIGN AND METHODS: The expression levels of apoptosis regulatory proteins and spontaneous apoptosis were measured in primary AML samples by Western blot analysis and flow cytometry. To determine the role of CD34+ cells in apoptosis resistance, spontaneous apoptosis in serum-free conditions and apoptosis regulatory protein levels were measured in CD34+ and CD34- sorted cells from CD34+ primary AML samples. RESULTS: We show that CD34+ AML fractions are more resistant to apoptosis than are corresponding CD34- AML fractions, and that this is paralleled by higher Bcl-2, Bcl-xL, Mcl-1, Pgp and lower Bax expression levels. Interestingly, as the percentage of CD34 cells increased in the primary AML sample, so too did the apoptosis resistance in the corresponding CD34- fraction, which was reflected by an increasing anti apoptosis protein profile. INTERPRETATION AND CONCLUSIONS: The data show that the CD34+ fraction is more resistant to apoptosis than is the corresponding CD34- fraction and secondly that the AML as a whole is more apoptosis resistant with increasing CD34 percentage. PMID- 12745270 TI - Natural killer resistance of a drug-resistant leukemia cell line, mediated by up regulation of HLA class I expression. AB - BACKGROUND AND OBJECTIVES: Drug-resistant leukemia cells may exhibit cross resistance towards immunological effector mechanisms by alterations of apoptosis pathways. This is particularly relevant in allogeneic bone marrow transplantation for leukemia, where the graft-versus-leukemia effect acts on cells pretreated with cytostatic drugs. Here, we clarify the mechanism underlying cross-resistance of drug-resistant variants of the T-leukemia cell line CEM towards natural killer cells. DESIGN AND METHODS: We determined the sensitivity of different CEM sublines to natural killer (NK) cytotoxicity, and separately analyzed the components of the killing machinery by detection of granzyme B-induced caspase cleavage and HLA class I-dependent recognition mechanisms. Furthermore, we studied regulation of HLA class I expression comparing CEM with other cell lines. RESULTS: We found that CEM cells resistant to cytostatic drugs or CD95 were cross resistant towards NK cells from a variety of donors. Granzyme B-induced caspase and PARP cleavage in the sensitive and resistant cells were comparable, indicating that downstream apoptosis pathways were not altered in the drug resistant cells. HLA class I molecules were upregulated in the resistant cells, inhibiting NK cells at the level of killer/target recognition. HLA class I upregulation was not found in other leukemia cell lines. INTERPRETATION AND CONCLUSIONS: This is the first description of HLA class I-mediated NK cross resistance in drug-resistant cells. This finding may have a clinical impact since it may be considered as a possible reason for resistance to a graft-versus leukemia approach in allogeneic bone marrow transplantation PMID- 12745271 TI - High-dose therapy with autologous transplantation for Hodgkin's disease: the Bologna experience. AB - BACKGROUND AND OBJECTIVES: In this work we examine the characteristics and outcome of patients with Hodgkin's disease (HD) treated with high-dose therapy (HDT) and autologous transplantation at our Institute between 1982 to 2000. DESIGN AND METHODS: A retrospective analysis was performed examining patients' characteristics, prior chemotherapy regimens, pre-transplant disease status, HDT regimen, source of stem cells, time for hematopoietic recovery, complications of transplantation, response rates, overall survival (OS) and relapse-free survival (RFS). RESULTS: Ninety-seven patients with HD were treated and had estimated 10 year OS and RFS rates of 32% and 60%, respectively. Disease status (sensitive vs. refractory) before HDT was the most powerful predictive parameter for OS and RFS in both univariate and multivariate analyses. The rate of transplant-related mortality in the whole cohort was only 1% whereas the rate of second malignancies was 3%. INTERPRETATION AND CONCLUSIONS: Our results confirm that HDT with autologous transplantation is associated with a durable RFS in a remarkable proportion of HD patients and that the procedure has a very low global early and late toxicity. PMID- 12745272 TI - Gene abnormalities in multiple myeloma; the relevance of TP53, MDM2, and CDKN2A. AB - BACKGROUND AND OBJECTIVES: Disruption of either the p14ARF- mdm2- p53 or p16INK4A Rb1 pathways produces a breakdown of regulatory mechanisms and creates a gateway for tumorigenesis. Since the incidence and clinical implications of abnormalities of TP53, CDKN2A (encoding for p16 and p14) and MDM2 genes (chromosome 12) in multiple myeloma (MM) is not clear, we investigated allelic loss at the former two loci and gain at the latter locus in a series of 82 MM patients. DESIGN AND METHODS: Dual color fluorescence in situ hybridization (FISH) was applied to bone marrow samples to establish the incidence of changes at the above mentioned loci. The CDKN2A locus was tested using a probe which hybridizes to 9p21 and also targets the p15INK4B gene. RESULTS: FISH analysis revealed the presence of monoallelic TP53 deletions in 12% of patients. Ten percent of patients had hemizygous deletion at 9p21, while a further 8% had loss of 1 of 3 loci in the presence of trisomy 9. MDM2 amplification in the face of chromosome 12 diploidy was seen in 8%, while another 8% had trisomy 12 with an equivalent increase in signals for MDM2. Clinical correlations revealed that allelic loss of TP53 was the only factor associated with resistance to chemotherapy. The presence of 9p21 deletion was associated with an IgA isotype but none of the abnormalities had a significant influence on overall or event-free survival. INTERPRETATIONS AND CONCLUSIONS: P53 and CDKN2A (9p21) allelic loss and amplifications of the MDM2 gene are infrequent events in myeloma. The incidence of the latter two events was, however, higher than previously reported. Deletion of the TP53 gene predicted resistance to chemotherapy, highlighting its importance in this disease process. PMID- 12745273 TI - B-cell compartment as the selective target for the treatment of immune thrombocytopenias. AB - BACKGROUND AND OBJECTIVES: Rituximab is a chimeric anti-CD20 monoclonal antibody active against normal and malignant B cells. Treatment with rituximab is associated with the development of a severe (even if transient) B-cell depletion from peripheral blood and lymphatic tissues. These effects could be useful in autoimmune diseases in order to interfere with the production of pathologic antibodies. DESIGN AND METHODS: To investigate this, we treated 20 patients with rituximab 375 mg/m2 i.v. every 7 days for 4 times. These 20 patients all had active and symptomatic autoimmune thrombocytopenia that had relapsed or was refractory to standard therapies (15 had idiopathic thrombocytopenic purpura, 1 idiopathic thrombocytopenia and neutropenia, 2 thrombocytopenia and concomitant undifferentiated connective tissue disease, and 2 had thrombocytopenia and concomitant B-cell lymphoprolipherative disorders). Only treatment with steroids, if strictly necessary to maintain a safe number of platelets, was allowed during the period of rituximab administration, but only patients who reached steroid discontinuation (previously not possible) were considered responders. RESULTS: Treatment was well tolerated and no acute or delayed toxic events were recorded. Rituximab proved to be active in 13/20 patients, with 9 complete and 4 partial responses. In 10/13 (77%) the response (platelet level > 50x10(9)/L) was prompt, being achieved already after the first of the four planned infusions. After a median follow-up of 180 days (range: 60-480) 4 patients had relapsed. Age < or = 60 years was correlated with a better response rate (p=0.03). No correlation was observed between response and gender, time from diagnosis to treatment (< 12 vs > 12 months), total and CD20+ lymphocyte count, level of CD20 expression on B cells before the therapy and pharmacokinetics of the drug. INTERPRETATION AND CONCLUSIONS: Rituximab appears to be a promising immunotherapeutic agent for the treatment of autoimmune thrombocytopenias. PMID- 12745274 TI - Thrombin generation assessed as endogenous thrombin potential in patients with hyper- or hypo-coagulability. AB - BACKGROUND AND OBJECTIVES: The endogenous thrombin potential (ETP) represents the balance between pro- and anti-coagulant forces operating in plasma and can be used to investigate hyper- and hypo-coagulability. As a preliminary step to larger clinical studies we investigated the effect on ETP of phospholipids, tissue factor (TF) and residual platelets in frozen plasma. DESIGN AND METHODS: We investigated platelet-poor and platelet-rich plasmas from healthy subjects, patients on oral anticoagulants (OA), or with hemophilia and women on oral contraceptives (OC), chosen as examples of the normal, hypo- and hyper-coagulable states in which ETP has been reported to be impaired. RESULTS: Phospholipids had only a slight effect on ETP in all conditions except in women on OC, in whom the best diagnostic efficacy was observed at 0.5 microM. TF had only a slight effect in all conditions except hemophilia, in which an ETP impairment was observed only at low (1 pM) concentration. Residual platelets had considerable effects on ETP in frozen plasmas, but this was abrogated by filtration before freezing. ETP in platelet-rich plasma at 150x103/mm3 was similar to that obtained in filtered plasma with 1.5 microM phospholipids in healthy subjects, patients on OA and patients with severe hemophilia, but not in those with mild- or moderate hemophilia, where the ETP was higher in platelet-rich plasma. INTERPRETATION AND CONCLUSIONS: The results suggest that the method can be used for investigations on the clinical value of ETP. Platelet-rich and platelet-poor plasma are suitable testing materials. The latter should be filtered before freezing to minimize the effect of residual platelets. PMID- 12745275 TI - Allogeneic hematopoietic stem cell transplantation with reduced-intensity conditioning in acute lymphoblastic leukemia: a feasibility study. AB - BACKGROUND AND OBJECTIVES: Numerous studies have reported the feasibility of performing allogeneic hematopoietic stem cell transplantation with reduced intensity conditioning, although results in individual diseases are scarce, with no studies in patients with acute lymphoblastic leukemia (ALL). We sought to analyze the results of reduced intensity conditioning for allografts in adult patients with ALL. DESIGN AND METHODS: We report the results of a reduced intensity conditioning regimen followed by allogeneic hematopoietic stem cell transplantation in 27 adult patients with high-risk ALL who were included in four prospective studies. RESULTS: The median age was 50 years; 23 (85%) patients were beyond first complete remission, 44% were chemorefractory and 41% were Philadelphia chromosome positive. Donors were mismatched related donors or volunteer unrelated donors in 12 cases (44%). The incidence of grades II-IV acute graft-versus-host disease (GVHD) was 48%, and 13 of 18 evaluable patients (72%) developed chronic GVHD. Currently nine patients are alive, with a median follow up of 809 days (range, 381-1375). The 2-year incidence of transplant-related mortality was 23% (95% CI, 11% to 46%), and the 2-year probability of overall survival was 31% (95% CI, 12 to 48%), while the 2-year incidence of disease progression was 49% (95% CI, 33% to 72%). The 2-year incidence of disease progression in patients with and without GVHD was 35% (95% CI, 19% to 57%) and 70% (95% CI, 47% to 100%), respectively (p=0.05). INTERPRETATION AND CONCLUSIONS: This retrospective study suggests that allogeneic hematopoietic stem cell transplantation with reduced intensity conditioning might be a useful therapeutic option for some patients with ALL who are ineligible for standard myeloablative conditioning. However, this treatment modality needs to be evaluated in prospective trials, and should not be employed outside clinical studies. PMID- 12745276 TI - The effects of natural killer-cell depletion on ex vivo expansion of hematopoietic progenitor cells from umbilical cord blood. AB - BACKGROUND AND OBJECTIVES: The small total number of hematopoietic progenitor cells (HPC) in cord blood limited its use in adult recipients. Since the hematopoiesis might be controlled by both positive and negative factors, the finding of negative cellular components and thereafter depletion of them would be of importance for further expansion of HPC from cord blood in vitro. The role of natural killer cells (NK cells) in hematopoiesis remains unclear and needs to be elucidated. DESIGN AND METHODS: Cord blood mononuclear cells were co-cultured in a liquid culture system containing the hematopoietic cytokines interleukin (IL) 1, IL-3, IL-6, stem cell factor (SCF), granulocyte-monocyte colony-stimulating factor (GM-CSF) and G-CSF for a total of 20 days with or without depletion of NK cells. RESULTS: The percentage of CD34+ cells was significantly higher in the NK cell-depleted group at each time point (day 5, day 10, day 15, day 20). This finding was further confirmed by examination of functional HPC including CFU-GM and CFU-GEMM. The intracellular interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha content of CD56+ NK cells increased more than 10-fold after incubation with hematopoietic cytokines (day 4, day 8). Neutralization of IFNgamma or TNFalpha, present in liquid culture, also significantly increased CFU GM formation and CD34+ cell content. INTERPRETATION AND CONCLUSIONS: The results indicate that depletion of NK cells may be beneficial in producing hematopoietic progenitor cells from cord blood in ex vivo expansion systems. The effect of NK cell depletion on ex vivo expansion of hematopoietic stem cells should be further assessed by a repopulating cell assay in NOD/SCID mice. PMID- 12745277 TI - Hematologic passport for athletes competing in endurance sports: a feasibility study. AB - BACKGROUND AND OBJECTIVES: Strategies based on the use of upper thresholds of hemoglobin or hematocrit to detect blood doping in endurance sports have essentially failed to deter this malpractice. With the aim of establishing a more effective strategy, we analyzed the biological variations of hematologic parameters in professional athletes and investigated the possibility of defining subject-specific reference ranges that could distinguish between physiologic and abnormal variability. DESIGN AND METHODS: Hemoglobin concentration, hematocrit, reticulocyte count, serum ferritin and soluble transferrin receptor levels were sequentially evaluated in 923 professional football players. Using the analysis of variance we tested the effect of age, ethnicity, exercise modalities and training phases on hematologic parameters and then estimated components of variation. The significance of the difference between two measures was obtained from the distribution of the within-subject variance (the so-called reference change). Subject-specific reference ranges were centered around the individual mean value with dispersion based on the 95th percentile of the coefficient of variation distribution. RESULTS: A total of 2,506 hematologic determinations were made. Exercise modalities were found to have important effects on hematologic parameters. Hemoglobin and hematocrit values were higher at the beginning of the competition season, and then declined in well-trained athletes. Aerobic exercise was clearly associated with lower values, suggesting that marginally low hemoglobin and hematocrit values should physiologically be found in endurance sports. At least five determinations were required to define subject-specific reference ranges reliably. Considering athletes showing normal indices of red cell production (i.e., reticulocyte count and soluble transferrin receptor), the 95th percentile of the coefficient of variation distribution was lower than 5% for both hemoglobin and hematocrit. Increases exceeding 10% in these latter parameters should to be considered abnormal. Score systems capable of efficiently detecting non-physiologic increases in red cell production were developed. INTERPRETATION AND CONCLUSIONS: Using proper sequential determinations of hematologic variables subject-specific reference ranges can be defined for hemoglobin and hematocrit. Thus, the hematologic passport is feasible and might be employed to exclude athletes with non-physiologic increases in hemoglobin and hematocrit from competitions. The hematologic passport should be used within a global strategy to deter blood doping. PMID- 12745278 TI - Inherited thrombocytopenias: a proposed diagnostic algorithm from the Italian Gruppo di Studio delle Piastrine. AB - BACKGROUND: Although no epidemiological study has so far been performed, inherited thrombocytopenias are considered to be very rare based on the number of case reports in the literature. However, diagnosis of these disorders is often difficult and requires competences that are limited to specialized centers. We, therefore, suspect that inherited thrombocytopenias are underreported because their diagnosis is often missed. OBJECTIVES: The aim of this study was to develop a diagnostic algorithm that utilizes the simplest possible diagnostic tests and that can also be used in centers that are not highly specialized. METHODS: The basic features of the diagnostic algorithm (definition and classification of hereditary thrombocytopenias, laboratory tests, sequence of investigations) were discussed by the members of the Italian Gruppo di Studio delle Piastrine during two meetings held in October 2000 and 2001. The sources of information were literature and personal experience. The final proposed diagnostic algorithm was produced by the authors of this paper and approved during a third meeting in May 2002. PERSPECTIVES: A definite diagnosis for patients with hereditary thrombocytopenias serves many purposes. It is essential to define the prognosis of the patients and to identify the best therapeutic approach. It also defines the risk of transmitting the disorder to progeny and, in many cases, allows prenatal diagnosis. Finally, several pieces of evidence indicate that not all the genetic thrombocytopenias have yet been identified. Exclusion of known disorders by accurate investigation is, therefore, the starting point for the discovery of these new illnesses. PMID- 12745279 TI - Screening for hemochromatosis in a population with abnormal iron status. PMID- 12745280 TI - Possible association between reactive oxygen metabolites and karyotypic abnormalities in myelodysplastic syndromes. PMID- 12745282 TI - Significant potentiation of anticoagulation by flu-vaccine during the season 2001 2002. PMID- 12745281 TI - Thalidomide before autologous stem cell transplantation in VAD-refractory multiple myeloma patients. PMID- 12745284 TI - Unusual sequence of VDJ rearrangement revealed by molecular analysis in a patient with indolent lymphoma. AB - We report a unique case of indolent lymphoma with an unusual VDJ rearrangement. Polymerase chain reaction (PCR) analysis of bone marrow at the time of diagnosis was positive for both BCL-2/JH and CDRIII rearrangements. After treatment, the patient achieved complete remission (CR) with slow disappearance of both rearrangements (CDRIII and then BCL-2/JH). Subsequently, two new CDRIII rearrangements were detected in bone marrow, peripheral blood, and lymph node tissue. After this conversion, fluorescent activated cell sorting (FACS) analysis demonstrated monoclonal disease, suggesting that both CDRIII rearrangements originated from one cell. Histological evidence of a B-cell small lymphocytic lymphoma (B-SLL) infiltrate in the bone marrow became evident approximately 1 year after the two CDRIII rearrangements appeared. Direct sequencing revealed that one of the CDRIII sequences consisted of a VDVDJ rearrangement. This is the first report of such a rearrangement in a case of indolent lymphoma. This type of rearrangement has been described to result from a secondary VDJ recombination in childhood acute lymphoblastic leukemia (ALL) leading towards oligoclonality and poorer prognosis. Our observations suggest that such a finding in an indolent lymphoma patient may precede transformation into an aggressive disease. Early detection by PCR could have substantial impact on the prognosis of such patients. PMID- 12745283 TI - T-cell clonality and myelodysplasia without chromosomal fragility in a patient with features of Seckel syndrome. AB - Seckel syndrome is a rare autosomal recessive disorder with characteristic craniofacial dysmorphism, skeletal defects, mental and prenatal growth retardation. About 50 cases have been reported in the literature. Hematologic abnormalities with associated chromosomal fragility have been noted in about 15% of the reported cases. We report a patient with Seckel syndrome with myelodysplastic features and clonal T-cells in the bone marrow but no evidence of chromosomal fragility. After 5 years of follow-up, this patient remains asymptomatic without any treatment and with stable peripheral blood counts. PMID- 12745286 TI - Sebastian syndrome: report of the first case in a South American family. AB - The Sebastian syndrome (SS) is a MYH9-related disorders, which are an extremely infrequent group of four autosomal dominant illnesses. SS consist of giant platelets, leukocyte inclusions and thrombocytopenia. To our knowledge, there are no case reports of this syndrome in South America. The propositus was a 35-year old Argentine woman with a history of purpuric lesions in her lower limbs and thrombocytopenia. Idiopathic thrombocytopenia purpura (ITP) was previously diagnosed, but she did not respond to treatment with steroids. Family history failed to provide any evidence of hearing loss, easy bruising, nephritis, renal failure or cataracts. The patient and 11 members of her family were evaluated. The diagnosis of SS was established by demonstrating giant platelets, thrombocytopenia and leukocyte inclusions in peripheral smear in two relatives and by peripheral smear and electronic microscopy in the propositus. MYH9-related disorders should be suspected whenever a patient has a low platelet count or a bleeding diathesis of unknown origin. In these cases, the history, carefully peripheral smear exam, immunocytochemistry and electronic microscopy will be of great help. Differentiation ITP with SS is important to avoid unnecessary diagnostic studies and treatments. PMID- 12745285 TI - Regression of metastatic osteosarcoma following non-myeloablative stem cell transplantation. A case report. AB - report the evidence of regression of multiple metastases following non myeloablative stem cell transplantation (NST) from an HLA-identical sibling in a case of relapsed fibroblastic osteosarcoma. The course of NST was well tolerated. Full donor chimerism was achieved on day +150 both for CD15+ and CD3+ cells. Complete remission was achieved on day +116. On day +210 the patient relapsed with a scapular metastasis that was unresponsive to four doses of donor lymphocyte infusion (DLIs). To our knowledge, this is the first reported case showing the achievement of complete remission following NST in an osteosarcoma patient. PMID- 12745288 TI - An identical mutation carried by different genes: Hb Frankfurt [alpha50(CE8)His >Gln]. PMID- 12745289 TI - Massive intestinal infiltration in a patient with bone T-cell rich diffuse large B cell lymphoma. PMID- 12745287 TI - Response to STI571 in chronic myelomonocytic leukemia with platelet derived growth factor beta receptor involvement: a new case report. AB - Based on its ability to inhibit the tyrosine kinase activity of ABL, as well as the c-kit and the Platelet Derived Growth Factor Receptor tyrosine kinases, the spectrum of diseases that may respond to STI571 is increasing. A recently recognized subgroup of myeloproliferative disorders/myelodysplastic syndromes (MPD/MDS) has a t(5;12)(q33;p13) with the activation of the gene for PDGFBR which encodes a receptor tyrosine kinase. Here, we present the case of a patient, with MPD/MDS, and eosinophilia, carrying a translocation t(5;12)(q33;p13) who achieved a complete remission following treatment with STI571, 400 mg daily. At the time of writing he still remains in complete remission with an excellent performance status. There is clearly a need for further studies of STI 571in MPD/MDS with chromosomal translocations involving PDGFBR to confirm this promising initial result. PMID- 12745290 TI - Transfusion of irradiated and stored RBCs to neonates. PMID- 12745291 TI - Frequency of factor VIII intron 1 inversion in a cohort of severe hemophilia A Italian patients. PMID- 12745294 TI - Cancer fatigue and its impact on patients: knowledge within the cancer care team. AB - Previous papers reported on the magnitude of fatigue as a problem for cancer patients and the growing recognition by patients, carers and health care professionals of the impact of cancer fatigue on quality of life. This paper presents findings from a United Kingdom survey of the knowledge of cancer fatigue and its impact on patients amongst cancer nurses, radiographers, haematologists and oncologists. We discuss how the context and chronology of care may influence differences in this knowledge between cancer care team members. Future directions for collaborative care, further research and interprofessional education are outlined. PMID- 12745295 TI - An evaluation of personal medical services: the times they are a changin'. AB - The Personal Medical Services (PMS) pilot sites, launched in England in 1997 by the Secretary of State for the then Conservative government, introduced a local contract for primary care, aimed at promoting flexibility, innovation and policy participation. As part of the National Evaluation of PMS, this paper considers the professional and organisational relationships established between service providers working in those PMS sites which specifically set out to address inequalities in access to primary care for vulnerable populations. The introduction of PMS enabled a change of cultural values in primary care, particularly regarding GPs' relationships with nurses and practice staff. However, PMS has not necessarily led to equal partnerships within primary care teams. Rather,in the selected sites evaluated new interprofessional relationships emerged. There was evidence of intra and interprofessional partnerships being forged, providing the basis for further improved intersectoral collaboration. There was also evidence that the GP based medical model made way for a community oriented/public health model with emphasis on health maintenance for the vulnerable. PMID- 12745296 TI - Of rocks and safe channels: learning to navigate as an interprofessional team. AB - A new interprofessional primary care team was established in a greenfield site. Using anethnographic approach, significant experiences in the formation of the team were documented and specific problem areas were defined and addressed using action research principles. The nature of the problems experienced and the processes used in their resolution are described and related to current policy recommendations and to existing literature on the theory and practice of team development. Problems include: the potentially varied expectations and managerial requirements of collaborating partners, the lack of clarity over professional roles and associated essential competencies, and the appropriate management and development of specialist professional roles. Conclusions drawn concern the complexity of the issues raised by interprofessional team working, and the value of existing techniques and models in this setting, such as the viable systems model, 360 degree appraisal and action research. PMID- 12745297 TI - Contradictions in interprofessional care: possibilities for change and development? AB - A descriptive model is outlined for analysis of contradictions in interprofessional care. Six persons from child psychiatric hospital staff and six hospital school teachers who worked with two teenagers were interviewed over a 7 month period in order to examine the nature of their occupational activity. The 47 interviews (225 pages of transcribed text) were examined using discourse analysis. The results indicate that the possibilities for development, which appear as contradictions in human activity and the efforts to resolve them, can be examined from the point of view of the evolution and change in semantic structures. PMID- 12745299 TI - Developing a virtual interdisciplinary research community in higher education. AB - As multidisciplinary collaboration in both clinical and research settings is becoming a key aspect of contemporary health care, strategies to enhance interprofessional interaction in postgraduate research programmes can offer important experiences to facilitate ongoing interprofessional relationships. This paper provides a retrospective appraisal of a strategy which used computer mediated communication to develop a virtual community network, known as'health_voice' accessed through a web page. The rationale for developing the network is presented, and the process of designing and establishing the web-site through an action research approach is described The outcome of the strategy is reviewed with regard to the relationships between the real' and 'virtual' community. Reflections on the developmental process contextualise the initiative within a concept of a community-of-practice. It is acknowledged that the use of a virtual arena for communication within a research community involves a cultural change in the dynamics of higher degree teaching and learning. Future plans to further embed the virtual environment within a postgraduate research culture are given. PMID- 12745298 TI - Student attitudes to undergraduate interprofessional education. AB - Interprofessional education in health care has been the focus of increasing attention in recent years. However, there is still great debate about when and how to introduce it in undergraduate studies. St George's Hospital Medical School with the Joint Faculty of Health Care Sciences of Kingston University was ideally placed to introduce, as part of its 1996 new curriculum, a Common Foundation Programme (CFP). This incorporated degree students in medicine, radiography, physiotherapy, and nursing learning together for the first term of their courses. As part of the evaluation of the CFP, students' attitudes to the course and each other were surveyed at the beginning and the end of the term, for the 1998 and 1999 intakes. The results showed that students arrive at university with stereotyped views of each other, and that these views appeared to become more exaggerated during the CFP. Students felt that the CFP would enhance interprofessional working, but there were concerns that it forced them to learn irrelevant skills. Students whose parents worked as health care professionals, held stronger stereotyped views. Our findings challenge any notion that students arrive without preconceived ideas about the other professions. Further work is needed to determine how best to break down stereotypes, and to advance our understanding of the most appropriate models for interprofessional education, to enable graduates to work effectively in today's environment. PMID- 12745300 TI - Interprofessional approaches to the prevention of health care errors. PMID- 12745305 TI - The National Psychiatric Morbidity Surveys of Great Britain--strategy and methods. AB - This paper describes the rationale and methodology of the first National Psychiatric Morbidity Surveys to be carried out in Great Britain. The objectives of the surveys were to estimate the prevalence of psychiatric morbidity among adults aged 16-64 living in Great Britain; to identify the nature and extent of social disabilities associated with psychiatric morbidity; to describe the use of health and social services by people with psychiatric morbidity and to investigate the association between mental illness and potential environmental risk factors in a household sample. Four separate surveys were carried out in order to meet the objectives; a private household sample (n = 10,108), a sample of institutions caring for the mentally ill (n = 1191), a sample of homeless people (n = 1166), and a supplementary sample of patients with psychosis living in private households (n = 350). A two-stage assessment procedure was used, in which all subjects were given the Revised Clinical Interview Schedule (CIS-R) administered by lay interviewers to assess neurotic symptoms and disorders and a psychosis screen, including the Psychosis Screening Questionnaire. Those who were positive on the psychosis screen were then interviewed by psychiatrists using the SCAN (incorporating the tenth edition of the Present State Examination). Large scale national surveys such as this augment the inadequate data on psychiatric morbidity that are routinely available and are, therefore, an important source of information upon which to base policy and generate aetiological hypotheses. These surveys provide a possible model for similar surveys in other countries. PMID- 12745306 TI - British Psychiatric Morbidity Survey. PMID- 12745307 TI - Building a picture of psychiatric morbidity in a nation: a decade of epidemiological surveys in Great Britain. PMID- 12745308 TI - The National Psychiatric Morbidity Surveys of Great Britain--initial findings from the household survey. AB - This paper describes the Household Survey from the National Survey of Psychiatric Morbidity. This covered a sample drawn at random from the population of Britain, with the exception of the Highlands and Islands of Scotland. The Postcode Address File was used as the sampling frame. Nearly 13,000 adults aged 16-65 were selected for interview, of which 10,108 (79.4%) were successfully interviewed. Eight percent could not be contacted and 13% refused interview. Psychiatric assessment was carried out by lay interviewers using the CIS-R. Subjects were also screened for psychosis, and screen-positive individuals were examined by psychiatrists using SCAN. Sixteen per cent of subjects scored above the standard cut-off of 12 on the CIS-R. The overall one-week prevalence of neurotic disorder was 12.3% in males and 19.5% in females. Unmarried and post-marital groups had high rates of disorder, as did single parents and people living on their own. Respondents in Social Class I had notably lower rates of neurotic disorder than the remainder of the sample. Unemployment was strongly associated with disorder. Subjects living in urban areas had a higher overall prevalence, but there was no significant variation by region. Black respondents had higher rates of disorders that were entirely explained by their age, family type, and social class. Individual neurotic disorders were all significantly commoner in women, with the exception of panic disorder. The one-year prevalence of functional psychoses was four per 1000, with no sex difference. Alcohol and drug dependence was considerably more prevalent in men. For the first time, the survey provides data on the prevalence and correlates of psychiatric disorder on a nationwide sample that can be used to inform equitable and effective national psychiatric services. PMID- 12745309 TI - Substance misuse and psychiatric comorbidity: an overview of the OPCS National Psychiatric Morbidity Survey. AB - There have been a number of national surveys of psychiatric morbidity, which have included questions on drugs, alcohol, and tobacco. These surveys have helped delineate the overlap between substance use and dependence and other psychological morbidity. There is a strong association reported between high substance consumption and other measures of psychological problems. This article provides an overview of a national household survey, a survey of institutional residents with psychiatric disorders, and a national survey of a homeless population. All three surveys used comprehensive and complex sampling strategies and lay interviewers to conduct structured diagnostic interviews. The household survey included over 10,000 households, the institutional survey interviewed 755 individuals, and the homeless survey of hostels, night shelters, day centres, and private sector leased accommodation interviewed 1,061 individuals. This overview looks at patterns of nicotine, alcohol, and other drug use in the different samples and examines interactions with other psychiatric morbidity. The survey reports that substance-related disorders are some of the commonest disorders in the community, with 5% of the household sample alcohol dependent, 7% alcohol dependent in the institutional sample and over 21% in the homeless sample recorded as alcohol dependent. Tobacco, alcohol and other drug use and dependence were dramatically higher in the homeless sample than in either of the other two samples. Substance use was significantly associated with higher rates of psychological morbidity as measured by the Clinical Interview Schedule Revised. Future service planning needs to take account of the striking disparity of prevalence of psychiatric disorders in different subsections of the population. PMID- 12745310 TI - Nicotine, alcohol and drug dependence, and psychiatric comorbidity--results of a national household survey. AB - There is a well-recognized relationship between substance use disorders and other psychiatric disorders. This relationship has been well documented in a range of population-based studies. This study aims to report on consumption patterns of and dependence on nicotine, alcohol, and non-prescribed drugs and to report on the levels of psychiatric morbidity in these groups. A national household study of psychiatric morbidity was conducted in England and Wales. Psychiatric assessment was based on the Clinical Interview Schedule-Revised (CIS-R). Measures of nicotine, alcohol and drug use and dependence were obtained. This paper compares the levels of psychiatric morbidity in the non-dependent and the nicotine-, alcohol- and drug-dependent cases. Twelve per cent of the non dependent population were assessed as having any psychiatric disorder compared with 22% of the nicotine-dependent, 30% of the alcohol-dependent, and 45% of the drug-dependent population. There is a clear relationship between dependence on nicotine, alcohol, and drugs and other psychiatric morbidity. PMID- 12745312 TI - Psychiatric morbidity among adults living in private households, 2000. PMID- 12745311 TI - Clarifying the relationship between unexplained chronic fatigue and psychiatric morbidity: results from a community survey in Great Britain. AB - The study examined the associations between several sociodemographic and psychosocial variables and unexplained chronic fatigue in the community before and after adjustment for psychiatric morbidity and determined the prevalence of fatigue and rate of disability resulting from fatigue in the general population. The study is a secondary analysis of 1993 data from a household survey of psychiatric morbidity conducted by the Office for Population Censuses and Surveys in Great Britain. The survey included 12,730 subjects age 16-64 years. Unexplained chronic fatigue was used as the dependent variable in a logistic regression analysis, with various sociodemographic and psychosocial variables and psychiatric morbidity as the independent variables. Psychiatric morbidity was assessed by using the Revised Clinical Interview Schedule. Fatigue was measured by using the fatigue section of the Revised Clinical Interview Schedule. A total of 10,108 subjects agreed to cooperate (79.4% participation rate). The prevalence of unexplained chronic fatigue was 9%. Subjects with psychiatric morbidity had higher rates of fatigue. Adjustment for psychiatric morbidity had a minor effect on the associations between sociodemographic factors and chronic fatigue. After adjustment, older subjects, women, and couples with children had higher rates of fatigue. Single subjects, widowed subjects, adults living with parents, and economically inactive subjects had lower rates of fatigue. Fatigue was associated with considerable disability, but the association between fatigue and psychiatric morbidity explained most of this disability. Unexplained chronic fatigue is a common condition, strongly associated with psychiatric morbidity. The close relationship between fatigue and psychiatric morbidity should not obscure the possibility of differences as well as similarities in their aetiologies. PMID- 12745313 TI - The influence of age and sex on the prevalence of depressive conditions: report from the National Survey of Psychiatric Morbidity. AB - Women are consistently reported to have a greater prevalence of depressive disorders than men. The reason for this is unclear, and is as likely to be social as biological. There is some evidence that the excess of depression is greater during women's reproductive lives. Data from the National Survey of Psychiatric Morbidity were used to test the hypothesis that the excess disappeared in the post-menopausal years and that obvious social explanations for this were inadequate. Subjects (n = 9792) from a random sample of the British population provided data for the analysis. Lay interviewers using the CIS-R carried out psychiatric assessment. Subjects with ICD-10 depressive episode or mixed anxiety/depression were compared with the remainder. Social variables that were likely to contribute to a post-menopausal decline in depressive disorders were controlled in logistic regression analyses. There was a clear reversal of the sex difference in prevalence of depression in those over age 55. This could not be explained in terms of differential effects of marital status, childcare, or employment status. This large and representative survey adds considerably to the increasingly held view that the sex difference in prevalence of depression is less apparent in later middle age. This may be linked to the menopause, and our attempts to explain it in terms of obvious conditions among social variables were not successful. More specific studies are required to clarify the finding. PMID- 12745314 TI - The circumstances of adults with a psychotic disorder. AB - This article presents some findings about adults with a psychotic disorder who were identified in the OPCS surveys of psychiatric morbidity. The main aim of the analysis was to identify characteristics associated with differences in the circumstances and health-related behaviour of adults aged 16-64 with psychosis. The analysis covers people who were identified by the various criteria used on the surveys as having a psychotic illness and who were considered to be living in private households. First we describe briefly the survey methods used and how diagnoses of psychosis were derived. We then present results from four topic areas covered in the analysis. These are: use of medication, use of health services, difficulties with activities of daily living, and social support. PMID- 12745315 TI - Socio-economic status, standard of living, and neurotic disorder. AB - Evidence on the association between socio-economic status and the prevalence of neurotic disorder is contradictory. We studied the association between three elements of socio-economic status and the prevalence of neurotic psychiatric disorder in a representative sample of adults aged 16-64 living in private households in the UK. A cross-sectional survey of 10,108 adults aged 16-65 resident in private households in the UK was selected by a multi-stage, clustered, random-sampling design. Neurotic disorders were defined using a standardised interview, the revised clinical interview schedule (CIS-R). Data for 9570 people were available for this study. We used housing tenure and access to cars as measures of standard of living; both were associated with the prevalence of neurotic disorder even after adjustment for other socio-economic and demographic variables, including the Registrar General's Social Class and educational attainment. Those people with no access to a car had an odds ratio for neurotic disorder of 1.4 (95% CI 1.1-1.7), compared with those who had access to two or more cars. People who rented their homes were also at increased risk (1.3 [1.1-1.5]). We estimated that about 10% of the neurotic disorder in the UK could be attributed to the increased prevalence of those without cars who rented their homes. There was a complex interaction between the Registrar General's Social Class and sex, and there was no independent association with educational attainment. There is an independent association between low standard of living and the prevalence of neurotic psychiatric disorder. The UK has experienced one of the largest increases in income inequality within western market economies over the past 20 years, and this inequality may have had adverse consequences for the mental health of the population. PMID- 12745316 TI - Urban-rural mental health differences in Great Britain: findings from the National Morbidity Survey. AB - Studies of urban-rural differences in prevalence of non-psychotic mental disorder have not given consistent findings. Such differences have received relatively little study in Great Britain. Data from 9777 subjects in the Household Survey of the National Morbidity Survey of Great Britain were analysed for differences between urban, semi-rural, and rural areas. Psychiatric morbidity was assessed by scores on the Revised Clinical Interview Schedule (CIS-R), together with alcohol dependence, drug dependence, and receipt of treatment from general practitioners. Associations with other characteristics were examined by logistic regression. Urban subjects had higher rates than rural of CIS-R morbidity, alcohol dependence, and drug dependence, with semi-rural subjects intermediate. Urban subjects also tended to be members of more deprived social groups, with more adverse living circumstances and greater life stress--factors themselves associated with disorder. Urban-rural differences in alcohol and drug dependence were no longer significant after adjustment for these factors by logistic regression, and differences on CIS-R morbidity were considerably reduced. There were no differences in treatment. There are considerable British urban-rural differences in mental health, which may largely be attributable to more adverse urban social environments. PMID- 12745317 TI - Neurotic disorders and the receipt of psychiatric treatment. AB - Access to psychiatric treatment by people with neurotic disorders in the general population is likely to be affected both by the severity of disorder and by sociodemographic differences. In the household component of the National Surveys of Psychiatric Morbidity >10,000 subjects in Great Britain with psychiatric symptoms were interviewed using the CIS-R. They were also asked about difficulties experienced in performing seven types of everyday activity. All subjects classed as having an ICD-10 disorder were questioned about their experience of treatment with antidepressants, hypnotics, and counselling or psychotherapy. Less than 14% of people with current neurotic disorders were receiving treatment for them. Within the previous year, only a third had made contact with their primary care physician for their mental problem: of these <30% were receiving treatment. Overall, 9% of people with disorders were given medication and 8% counselling or psychotherapy. A diagnosis of depressive episode was that most associated with antidepressant medication. Treatment access was affected by employment status, marital status, and age, but the major determinant was symptom severity. Neither sex nor social class influenced which people received treatment. People with psychiatric disorders seldom receive treatment, even when they have consulted their primary care physician about them. In many cases, this must represent unmet needs with a strong claim on health resources. There are also inequalities in the receipt of treatment, although the major influence is the severity of disorder. PMID- 12745318 TI - Unequal access and unmet need: neurotic disorders and the use of primary care services. AB - In this paper we use data from the National Survey of Psychiatric Morbidity to examine how many people with neurotic disorders receive professional evaluation, and how this is affected by clinical and sociodemographic differences. We hypothesized that psychiatric symptoms and attendant dysfunctions would both have an effect on contacting, and that key demographic variables would not. The household component of the British National Surveys of Psychiatric Morbidity was based on a random sample of >10,000 subjects. Lay interviewers using the CIS-R established psychiatric symptoms and ICD-10 diagnosis. Social dysfunction was tapped by asking about difficulties in performing seven types of everyday activity. We examined symptom score, ADL deficit score, and demographic variables in relation to contact with primary care physicians for psychiatric symptoms. The major determinant of contacting a primary care physician was severity, mainly due to the level of psychiatric symptoms, but with an independent contribution from social dysfunction. There were also significant contributions from sex, marital status, age, employment status, and whether the subject had a physical condition as well. The major influence on whether people seek the help of their family doctors for mental health problems is the severity of disorder. Although there are some social inequalities in access to family doctors, these are less important. The most salient finding from our study is that even people suffering from high levels of psychiatric symptoms very often do not have contact with professionals who might help them. PMID- 12745319 TI - The reluctance to seek treatment for neurotic disorders. AB - In previous papers from the National Survey of Psychiatric Morbidity in Great Britain, we have demonstrated that people with neurotic disorders rarely present their symptoms to primary care physicians and when they do, are quite likely not to be given treatment. In this paper, we examined survey respondents' reports of specific instances of reluctance to seek help in relation to sociodemographic, socio-economic and clinical attributes of our subjects. All people in the National Household Survey assessed as having a neurotic disorder were asked if at any time in the previous year they had avoided seeking appropriate treatment. Clinical measures included diagnosis, symptom severity, and deficits in carrying out tasks of daily living. Of nearly 1400 respondents, a quarter said they had not been to see a doctor at some time in the past year when they or their family felt they should have. The major determinant of this reluctance was symptom severity: more severe cases were more likely to report an episode of reluctance. Reasons included those related to ignorance of neurotic disorders and the effectiveness of treatment and to stigma. The attitudes detected in our subjects with neurotic disorder help to explain why people do not always seek effective help for their mental disorders, and are indicators of a worrying public education gap that will be hard to bridge. PMID- 12745321 TI - The prevalence of psychiatric morbidity among homeless adults. PMID- 12745320 TI - The prevalence of psychiatric morbidity among adults living in institutions. AB - This paper presents prevalence data from the 1994 OPCS survey of psychiatric morbidity among adults permanently resident in institutions catering for people with mental health problems in Great Britain. It describes briefly the survey methods used, and how diagnoses of psychiatric morbidity were derived. Its main aim is to show the prevalence of psychiatric morbidity in different types of institutional settings. Residents were eligible for the survey if they were aged 16 to 64 at the date of sampling and were permanently resident at the establishment. Residents were defined as permanently resident if they had been living in the sampled establishment for six months or more, or had no other permanent address, or were likely to stay in the establishment for the foreseeable future. In 1994, about 33,200 adults aged 16 to 64 were permanently resident in accommodation for people with mental health problems. About a third of residents were in NHS hospitals, while about two-thirds were in residential care facilities. About two-thirds of adults interviewed suffered from schizophrenia, delusional and schizoaffective disorders. About 8% suffered from neurotic disorders and 8% suffered from affective psychoses (mainly bipolar affective disorder). The prevalence of schizophrenia, delusional, and schizoaffective disorders was higher in hospitals than in residential care, while the prevalence of neurotic and related disorders was higher in residential accommodation. The prevalence of schizophrenia, delusional, and schizoaffective disorders was higher in NHS psychiatric hospitals and general hospital units than in private hospitals, clinics or nursing homes. PMID- 12745322 TI - Survey of the health and well-being of homeless people in Glasgow. PMID- 12745323 TI - Psychiatric morbidity among young offenders in England and Wales. PMID- 12745324 TI - Non-fatal suicidal behaviour among prisoners. PMID- 12745325 TI - Substance misuse among prisoners in England and Wales. PMID- 12745326 TI - Psychiatric morbidity among women prisoners in England and Wales. PMID- 12745327 TI - Prevalence of pervasive developmental disorders in the British nationwide survey of child mental health. AB - The prevalence of pervasive developmental disorders (PDD) is not well established and needs monitoring. We investigated the prevalence of PDDs in the 1999 nationwide British survey of child and adolescent mental health. A randomized stratified sample of children (n = 12,529) aged 5-15 was generated from Child Benefit Records. Trained interviewers interviewed parents and youths aged 11 or older with a standardized diagnostic interview (Development and Well-Being Assessment) and questionnaire data (Strengths and Difficulties Questionnaire) were obtained from teachers and parents who also completed self-report measures of psychological distress. A team of experienced clinicians using all data sources achieved final diagnostic determination. A total of 10,438 (83%) interviews were conducted. There were two girls with Rett syndrome (weighted prevalence: 3.8/10,000 girls) and 27 children with other PDDs (weighted prevalence: 26.1/10,000). Compared to children with a psychiatric disorder other than PDD, social but not behavioural problems were more frequent in the PDD group. Parents of children with PDDs had higher rates of psychological distress than those from the two comparison groups. Consistent with other recent surveys, PDD rates are higher than those reported 30 years ago. The burden associated with PDDs is very high. PMID- 12745328 TI - Using the Strengths and Difficulties Questionnaire (SDQ) to screen for child psychiatric disorders in a community sample. AB - Child psychiatric disorders are common and treatable, but often go undetected and therefore remain untreated. To assess the Strengths and Difficulties Questionnaire (SDQ) as a potential means for improving the detection of child psychiatric disorders in the community, SDQ predictions and independent psychiatric diagnoses were compared in a community sample of 7984 5-15 year olds from the 1999 British Child Mental Health Survey. Multi-informant (parents, teachers, older children) SDQs identified individuals with a psychiatric diagnosis with a specificity of 94.6% (95% Cl, 94.1-95.1%) and a sensitivity of 63.3% (59.7-66.9%). The questionnaires identified over 70% of individuals with conduct, hyperactivity, depressive, and some anxiety disorders, but fewer than 50% of individuals with specific phobias, separation anxiety and eating disorders. Sensitivity was substantially poorer with single-informant rather than multi-informant SDQs. Community screening programmes based on multi-informant SDQs could potentially increase the detection of child psychiatric disorders, thereby improving access to effective treatments. PMID- 12745329 TI - The Strengths and Difficulties Questionnaire: a pilot study on the validity of the self-report version. AB - The self-report version of the Strengths and Difficulties Questionnaire (SDQ) was administered to two samples of 11-16 year olds: 83 young people in the community and 116 young people attending a mental health clinic. The questionnaire discriminated satisfactorily between the two samples. For example, the clinic mean for the total difficulties score was 1.4 standard deviations above the community mean, with clinic cases being over six times more likely to have a score in the abnormal range. The correlations between self-report SDQ scores and teacher--or parent rated SDQ scores--compared favourably with the average cross informant correlations in previous studies of a range of measures. The self report SDQ appears promising and warrants further evaluation. PMID- 12745330 TI - Prevalence of obsessive-compulsive disorder in the British nationwide survey of child mental health. AB - Obsessive-compulsive disorder (OCD) is a disorder that appears to be under diagnosed and under-treated, despite the evidence for effective treatments. There are variable estimates of OCD prevalence in the under-16s and published rates give little indication of age trends. The aim of this study was to establish the prevalence and associates of OCD in young people aged 5-15 years. The method was a nationwide (UK) epidemiological study of rates of psychiatric disorder in 5-15 year olds (1999 British Child Mental Health Survey): 10,438 children were assessed. Twenty-five children with OCD were identified (weighted overall prevalence 0.25%; 95% CI 0.14-0.35), with prevalence rising exponentially with increasing age. Compared with normal controls, children with OCD were more likely to be from lower socio-economic class and of lower intelligence. Only three of these children had been seen by specialist children's services. Although OCD is rare in young children, the rate increases towards the adult rates at puberty. Children with OCD have additional psychosocial disadvantage. The majority of the childhood cases identified in this survey appear to have been undetected and untreated. PMID- 12745332 TI - Making psychiatric epidemiology useful: the contribution of epidemiology to government policy. AB - This paper aims to discuss the contribution of epidemiology to aspects of public policy that have either a direct influence on mental health and mental disorders, or an indirect effect by influencing environmental factors which influence mental health. Both kinds of public policy will need to be considered by governments wishing to protect, promote, and improve the mental health of their populations. The paper draws on information from both relatively wealthy and low-income countries. The paper defines epidemiology and mental health policy, sets out the range of government policies which may have an impact on mental health, and explores the ways in which epidemiology may contribute to mental health policy in relation to service inputs, processes and outcomes as well as to wider government policies. The paper also examines the advantages and disadvantages of different sources of data. There are a number of reasons to carry out large-scale surveys of psychiatric morbidity. First, effective policy should address the needs of the population, which can best be assessed by the epidemiology and the social and economic causes and consequences of psychiatric morbidity. Secondly, representative information in a defined geographic area can document the use of existing services and can estimate the extent of unmet needs and the services required meeting those needs. Thirdly, valid information on prevalence and associated risk factors of presumed causal importance allow aetiological hypotheses to be generated and tested and models developed for prevention. Finally, by repeating community surveys, it is possible to monitor the health of the population and trends. Epidemiological findings emphasize the importance of mental health policy addressing the key role of primary care, the social context and social consequences of disorder, the importance of addressing services for children, the need to reduce premature mortality from suicide and from physical illness. Epidemiological findings show that mental health and mental disorders are related to the environment both in its structural physical sense and in the sense of the social processes connected to and influenced by particular settings. Thus epidemiology can contribute to general policies on employment and unemployment, housing and homelessness, education, and women's issues. Mental health policy is increasingly recognized as an essential area for countries wishing to enhance their economic, social, and human capital. Epidemiological data are a basic prerequisite to informing such policies. Expert professional and epidemiological advice to ministries is essential if policy is to be rooted in the evidence for population needs, risk factors, effective treatments and services, and measurement of outcomes. It is therefore important to develop the capacity for policy work in the psychiatric profession by including public health, epidemiology, and policy placements for young psychiatrists. PMID- 12745331 TI - Mental health of children and adolescents in Great Britain. AB - The findings described in this report and summarized here focus on the prevalence of mental disorders among 5-15 year olds and on the associations between the presence of a mental disorder and biographic, sociodemographic, socio-economic, and social functioning characteristics of the child and the family. Causal relationships should not be assumed for any of the results presented in this report. PMID- 12745333 TI - Sanitation and hygiene in urban and rural households in East Africa. AB - Latrine possession, disposal of children's faeces and waste-water in 1015 households in 33 sites in Kenya, Tanzania and Uganda were studied in 1997. Assistants conducted interviews and observed the state and use of latrines, disposal of children's faeces, wastewater, and household socio-demographic characteristics. Latrine possession was 92.4% in Uganda, 95% in Kenya and 99.5% in Tanzania. In unpiped sites, 73.5% of Ugandan, 90.5% of Tanzanian and 95% of Kenyan households had latrines. Over 30% of latrines in rural Uganda were contaminated with faeces, compared with 10% in Tanzania. More latrines in urban Kenya and Uganda had contaminated surroundings than in the rural areas. The mean number of people using a toilet in the urban areas (10) was significantly higher than in rural areas (7), (F = 45.5; P < 0.001). Toilets in Kenya and Uganda were more likely to be fouled than in Tanzania. Households where the head was an educated professional or business person, or the toilet had a door, lid or concrete wall or floor or waste water was disposed of in the latrine, were less likely to have fouled toilets. Most households disposed of the faeces safely with a few placing them in the garden or elsewhere. The study emphasises the need to promote appropriate sanitation and hygiene. PMID- 12745334 TI - The disposal of used sharps by diabetic patients living at home. AB - The disposal of sharps generated in the community has been identified as an area of public health and environmental health concern. While there is a large amount of literature on sharps disposal practices in healthcare settings, the sharps disposal practices of diabetic patients living at home has been poorly documented. This study describes the sharps disposal practices of diabetic patients in South Staffordshire, an English health district. A randomly selected sample of 1,348 adult (aged >or= 16 years) diabetic patients were obtained from the district population-based diabetes register. A self-administered questionnaire was posted to the sample. Non-responders received up to two reminders. A response rate of 91% was achieved. Household containers were used by: 34.1% of respondents for syringes; 35.1% for lancets; and 27.6% for needles. Sharps boxes were the least used method of sharps disposal. Many respondents indicated that they had received only verbal information on how to dispose of their sharps. Those who recalled receiving information were more likely to dispose of their sharps safely. The results of this study suggest that sharps are disposed of in the most convenient manner, into the household waste. This contributes to environmental pollution and places people at risk of physical and psychological trauma. PMID- 12745335 TI - Role of environmental pollutants on immune functions, parasitic infections and limb malformations in marine toads and whistling frogs from Bermuda. AB - Soil, water, and amphibian tissues collected between 1995 and 1999 from 15 study sites in Bermuda were analysed for pesticides and heavy metals. The most abundant pesticide residue in soil was p,p'-dichlorodiphenyldichloroethylene (DDE) which was found at all sites in concentrations ranging from 0.003 to 4.023 p.p.m. No pesticide residues were found in water. DDE was also recovered from the livers and fat bodies of marine toads (Bufo marinus) and whistling frogs (Eleutherodactylus johnstonei). Analyses of food sources consumed by these anuran species revealed residue levels of p, p'-DDE ranging from 0.05 to 0.217 p.p.m. Other soil residues included dichlorodiphenyltrichloroethane (DDT) at eight study sites, Dicofol(kelthane) at eight sites, dieldrin at five sites, and polychlorinated biphenyls (PCBs) as Arochlor 1254 and Arochlor 1260 at seven sites. Analyses of toad livers revealed significant concentrations of cadmium, chromium, copper and zinc. Livers of Bermuda toads exhibited altered hepatocytic morphology and an increased number of melanomacrophages and possible granulomas, while spleens showed a marked decrease in white pulp. Spleen cells from Bufo marinus collected at one site having high levels of cadmium exhibited a decreased B cell response to lipopolysaccharide. The incidence of trematode infection in Bufo marinus increased from 53.8% in 1995 to 90% in 1999. Deformity rates in the limbs of subadult and adult toads ranged between 15 and 25%. Examination of 1,995 newly-metamorphosed toads revealed deformity rates as high as 47%. The current comprehensive study suggests that environmental pollutants may account for immunosuppression, increased susceptibility to infections, limb malformations and possible decline in amphibian populations from Bermuda. PMID- 12745336 TI - The application of a facet scale job satisfaction model for environmental health officers in Australia and Scotland. AB - The direction provided from published research emphasises that job satisfaction is multi-dimensional, and the associations with work related and non-work related predictor variables as being complex. The isolation of the discrete influence of individual predictor variables is therefore difficult. The application of multivariate analysis incorporating hierarchical stepdown protocols and interaction effects offers one approach that provides for the discrimination between the explanatory characteristics of these variables. Principal Axis Factoring with oblimin rotation and Structural Equations Modelling were applied to survey data to develop a Measurement Model comprising six latent constructs that characterised the job satisfaction of Environmental Health Officers (EHOs) in Australia. The model was successfully applied to data from the EHOs in Scotland. This paper presents the application of the Measurement Model to examine the association between selected work content and the demographic aspects of the EHOs in Australia and in Scotland, and their job satisfaction. The analysis protocol included Canonical Correlation, Multiple Linear Regression and Factorial Manova and Mancova. The unique variance explained by the work content factors and the demographic aspects of the job were similar. The linear representation of model pathways provided more comprehensive explanatory power than the curvilinear association. The presentation of evidence of multidimensional complexity, including interaction effects, underscored the need for caution before generalising on the influence of individual factors on job satisfaction. The analysis protocol presented in this paper can be applied to other situations in environmental health involving multivariate complexity. PMID- 12745337 TI - Effect of aerobic and anaerobic digestion on the viability of Cryptosporidium parvum oocysts and Ascaris suum eggs. AB - The viability of Cryptosporidium parvum oocysts and Ascaris suum eggs inoculated into aerobic and anaerobic digesters was measured. The digesters were maintained at 37 degrees C, 47 degrees C, and 55 degrees C, with 10-day detention times. Eggs and oocysts were added to each digester in a single spike or in chambers placed in the digesters for varying periods. Oocysts were inactivated very rapidly in all systems as determined by a dye permeability assay, > 99% inactivated after 10 days at 37 degrees C, 4 days at 47 degrees C, and 2 days at 55 degrees C. Eggs were more rapidly inactivated in anaerobic digesters than in aerobic digesters. At 55 degrees C, eggs in both anaerobic and aerobic digesters were > 99% inactivated within 1 h. At 47 degrees C, anaerobic digestion inactivated around 95% eggs in 2 days, but around 25% of the eggs were still viable after 10 days in aerobic digesters. At 37 degrees C, anaerobic digestion inactivated more than 75% of the eggs after 10 days, but in the aerobic digester at 37 degrees C, 10 days of treatment had no effect on viability. The oocysts and eggs added in chambers appeared to behave similarly to these pathogens added directly to the biosolids within the digesters. PMID- 12745338 TI - A theoretical approach to assess microbial risks due to failures in drinking water systems. AB - A failure in treatment or in the distribution network of a surface water-works could have serious consequences due to the variable raw water quality in combination with an extended distribution. The aim of this study was to examine the theoretical impact of incidents in the drinking water system on the annual risk of infection in a population served by a large water treatment plant in Sweden. Reported incidents in the system were examined and a microbial risk assessment that included three pathogens, Cryptosporidium parvum, rotavirus and Campylobacter jejuni, was performed. The main risk incidents in water treatment were associated with sub-optimal particle removal or disinfection malfunction. Incidents in the distribution network included cross-connections and microbial pollution of reservoirs and local networks. The majority of the annual infections were likely to be due to pathogens passing treatment during normal operation and not due to failures, thus adding to the endemic rate. Among the model organisms, rotavirus caused the largest number of infections. Decentralised water treatment with membranes was also considered in which failures upstream fine-pored membranes would have little impact as long as the membranes were kept intact. PMID- 12745339 TI - Water - health relationships in developing countries: a case study in Tulkarem district in Palestine. AB - This study focuses on quantitative and qualitative determinants of drinking water in the Tulkarem district of Palestine, and the effect of water pollution on the health of its inhabitants. Five hundred drinking water samples were obtained from the records of the Public Health Department/Ministry of Health (MoH) at Tulkarem district for the year 1999, which were collected from different sources in the district. The samples were examined for free chlorine residual concentration, total coliforms and faecal coliforms. Data of water-related diseases was obtained from the records of public health clinics in the district. Many of the examined samples were unacceptable according to the Palestinian and WHO standards. It was found that (60.6%) of the samples have concentrations of free chlorine residual less than 0.2 ppm, which is the minimum concentration, recommended by WHO. Out of these samples, 34% and 9.2% were contaminated with total coliforms and faecal coliforms respectively. It was obvious that the prevalence of water-related diseases is much higher in the areas with contaminated or nonchlorinated drinking water than in other areas. PMID- 12745340 TI - PM(10) and its impacts on health - a case study in Mumbai. AB - Air pollution in urban areas is a major health concern. Air pollution in cities has been linked to increased rates of mortality and morbidity in developed and developing countries. There is consistent evidence that the levels of fine particulate matter in the air are associated with risk of death from all causes of cardiovascular and respiratory illness. The present study predicts the health impact due to PM(10) (respirable particulate matter) in an industrial, residential and commercial area of Mumbai city. The estimates of the impacts on health is mainly based on Ostro's approach. PMID- 12745341 TI - The use of an air filtration system in podiatry clinics. AB - A small-scale study was conducted to ascertain the efficiency and effectiveness of an air filtration system for use in podiatry/chiropody clinics (Electromedia Model 35F (A), Clean Air Ltd, Scotland, UK). Three clinics were identified, enabling comparison of data between podiatry clinics in the West of Scotland. The sampling was conducted using a portable Surface Air Sampler (Cherwell Laboratories, Bicester, UK). Samples were taken on two days at three different times before and after installation of the filtration units. The global results of the study indicate the filter has a statistically significant effect on microbial counts, with an average percentage decrease of 65%. This study is the first time, to the authors' knowledge, such a system has been tested within podiatric practice. PMID- 12745342 TI - Lead exposure in day care centres in the Caracas Valley--Venezuela. AB - Lead has been used as an additive to improve the anti-knock quality of gasoline. However, the combustion of lead gasoline produces particles rich in this element. These kinds of particles impact the alveoli and move into the bloodstream, causing serious health problems. The objective of this study is to determine the granulometric distribution, mineralogy, and lead levels in samples such as settled dust and soil, as well as lead concentration in drinking water, in different day care centers located in the metropolitan area of Caracas, which have been previously studied and classified, according to the Total Suspended Particles (TSP) levels by the national environmental office, as high (80 microg m(-3)) and low (50 microg m(-3)) polluted areas. The chemical analysis was performed by atomic absorption spectroscopy and atomic emission spectroscopy inductively coupled plasma. The mineralogy was determined using X-ray differentiation. Results show that all samples are enriched by small particles (<44 microm). Lead concentrations in day care centres located in areas of high vehicular density were higher than those in areas of low vehicular traffic. Lead concentration for drinking water samples was below the standard value reported by the Environmental Protection Agency (EPA) (15 microg Pb/l). PMID- 12745343 TI - The knowledge of healthcare professionals with regard to the treatment and prevention of head lice. AB - This study examined the knowledge and practice of healthcare professionals regarding the prevention and treatment of head lice 18 months after dissemination of local guidelines. A self-administered postal questionnaire was sent to all primary healthcare professionals (general practitioners and practice nurses), community healthcare professionals (community paediatricians, health visitors and school nurses) and pharmacists in South Staffordshire health district, UK. The overall response rate was 48% (range 24-63%). Compared to other groups: community healthcare professionals were more likely to refer to the guidelines and have adequate knowledge of treatment and prevention; pharmacists were least likely to refer to the guidelines; and primary healthcare professionals were most likely to have poor knowledge about prevention and treatment. Overall, healthcare professionals' knowledge regarding prevention methods was significantly better than their knowledge of treatment methods (63% vs. 5%, P <0.00001). In conclusion, the results of this study suggest that healthcare professionals' knowledge of control methods for head lice varies widely and is sub-optimal and may contribute to ineffective head lice control. PMID- 12745344 TI - Bacteriological and chemical quality of swimming pools water in developing countries: a case study in the West Bank of Palestine. AB - Monitoring was carried out during summer 2000 in all the swimming pools in the West Bank of Palestine. Fifty-eight water samples, collected from 46 swimming pools, were examined for Coliforms and bacterial species including Streptococci, Salmonellae, and Staphylococcus. Salmonellae were isolated in 21 out of 23 samples. All of the examined samples from the swimming pools water were unacceptable according to the Palestinian and WHO standards. Extensive efforts are required to improve the water quality of the swimming pools in the West Bank, mainly public awareness, training of governmental inspectors, operators and owners of the swimming pools, in addition to a strict system for monitoring of the water quality. PMID- 12745346 TI - The microbiological quality of ready-to-eat foods with added spices. AB - A microbiological study of ready-to-eat foods with added spices or spice ingredients was undertaken to identify any risk factors in the production, storage and display of this product and to establish their effect on microbiological quality. Examination of 1946 ready-to-eat foods from sandwich bars, cafes, public houses, restaurants, specialist sandwich producers, bakers, delicatessens, market stalls and mobile vendors found that 1291 (66%) were of satisfactory/acceptable microbiological quality, 609 (32%) were of unsatisfactory quality, and 46 (2%) were of unacceptable quality. Unacceptable results were due to high levels of B. cereus and/or other Bacillus spp. (>/=10(5) cfu g(-1)). Unsatisfactory results were mostly due to high Aerobic Colony Counts (up to >/=10(7) cfu g(-1)), Enterobacteriaceae (>/=10(4) cfu g(-1)), Escherichia coli (>/=10(2) cfu g(-1)), and Bacillus spp (>/=10(4) cfu g(-1)). Examination of 750 spices and spice ingredients revealed that B. cereus were present in 142 (19%) samples, other Bacillus spp. in 399 (53%) samples, and Salmonella spp. (S. enteritidis PT 11) in one (<1%) sample. Approximately a third (222) of spice and spice ingredients examined contained high counts (>/=10(4) cfu g(-1)) of B. cereus and/or other Bacillus spp., and appeared to be associated with the corresponding ready-to-eat foods containing similar high counts of these organisms (P<0.0001). Acceptable microbiological quality of ready-to-eat foods to which spices or spice ingredients have been added was associated with premises that had management food hygiene training and hazard analysis in place. Poor microbiological quality was associated with preparation on the premises, premises type, little or no confidence in the food business management of food hygiene, and small premises as indicated by local authority inspectors' confidence in management and consumer at risk scores. PMID- 12745345 TI - Occupational exposure to cholinesterase inhibiting pesticides: a Greek case. AB - The determination of plasma or serum cholinesterase is absolute and it is considered as a reliable index of exposure in workers of organophosphorus pesticides industries. In the present study the plasma cholinesterase of 28 persons working in the packaging of an ortho-thio-phosphate was determined, before and after their exposure to this agent. The results of this study showed a plasma cholinesterase depression of 37%, a decrease which was statistically significant (P<0.001). PMID- 12745347 TI - Characterization of drug-resistant Staphylococcus aureus isolated from poultry processing plants in Western Australia. AB - Poultry processing plants can provide a favourable environment for the survival and transmission of Staphylococcus aureus. It is known that infections due to antibiotic-resistant strains of S. aureus are an increasingly serious problem clinically and, since antibiotic exposure in food-animal species may lead to antibiotic-resistant bacteria, it is possible that processed poultry may constitute a reservoir for disseminating antibiotic-resistance into the community. The present study was undertaken to determine the prevalence of antimicrobial-resistant S. aureus in two poultry processing plants, and to characterize the isolates by antimicrobial susceptibility and chromosomal and plasmid DNA analysis. One hundred and twenty-six S. aureus were isolated from two poultry processing plants in Western Australia. All were sensitive to 14 of the 26 antimicrobials tested and all isolates were resistant to at least one antibiotic and one chemical marker, the prominent resistance combination being to penicillin and cadmium (89%). Forty-six (36.5%) of the isolates were resistant to six or more of the antimicrobial agents tested. Overall there were no consistent resistance patterns for the isolates and no consistent patterns were found between and within the two processing plants. There were 24 epidemiologically unrelated Sma1 contour-clamped homogeneous electric field (CHEF) groups and 17 different plasmid profiles detected among the isolates. All isolates were found to harbour from between one to seven plasmids. The majority of isolates carried at least one large plasmid (22-48 Kb), and one or more small plasmids (1-3 Kb). Some isolates with epidemiologically related CHEF patterns had similar plasmid profiles and resistance patterns. PMID- 12745348 TI - Immunologic and other biological parameters as a function of smoking status and of residence in areas differing in terms of air pollution. AB - In the industrial region of La Louviere (Belgium), healthy persons, presenting at a consultation of preventive medicine, showed differences as a function of smoking and of residence in one of four areas differing in sources of emissions, population density and green zones. Smokers had significantly (P/=2.25, significantly so for the Power Station-Landfill area. A non-significant but consistent trend towards higher lymphocyte and CD8 lys counts and higher complement C3c serum levels further suggested an association between residence in a more polluted area and immunologic features. The number of CD3+CD56+ lys microl(-1), reported to be increased in cancer patients, showed a negative correlation with the CD4/CD8 ratio (r(2)=0.132, P<0.0001). PMID- 12745349 TI - Multiple logistic regression modelling substantiates multifactor contributions associated with sick building syndrome in residential interiors in Mauritius. AB - This paper presents a mathematical model that depicts the relationship between the possibility of occurrence of common health problems and factors leading to Sick Building Syndrome symptoms in domestic interiors in Mauritius. The prevalence of upper respiratory symptoms (dry eyes, runny nose), central nervous system symptoms (headache, nervousness), and musculoskeletal symptoms (pain/stiffness in shoulders/neck) were found to be elevated when responses were statistically regressed to type of building and age of respondents. The model presented here will be useful in helping to identify and quantify the relative role of factors that contribute to Sick Building Syndrome. Thus it may be possible to evaluate the effectiveness of current building operation practices and to prioritise allocations of resources for reduction of risk associated with Indoor Environmental Air Quality. PMID- 12745350 TI - Daily variations in effluent water turbidity and diarrhoeal illness in a Russian city. AB - To assess an association between temporal variations in drinking water quality and gastrointestinal (GI) illness, a cohort study involving 100 randomly selected families (367 individuals) was conducted in the city of Cherepovets, Russia from June through November 1999. Participants maintained daily diaries of gastrointestinal symptoms, water consumption and other behavioural exposure variables, while daily effluent water quality data were provided by the water utility. The cumulative incidence rate of self-reported gastrointestinal diseases, 1.7 cases per person-year, was almost two orders of magnitude higher than that of officially reported GI infections in the city. An interquartile range increase in effluent water turbidity of 0.8 Nephelometric Turbidity Units was associated with a relative risk of self-reported GI illness of 1.47 (95% Confidence Interval 1.16, 1.86) at a lag of 2 days after control for daily rate of consumption of non-boiled tap water, behavioural covariates, day of the week and a seasonally-related linear trend. In the analysis by subsets of study participants stratified by non-boiled tap water consumption, no statistically significant associations between turbidity and GI illness were found for the study participants who always boiled their drinking water. For individuals who drank non-boiled tap water, statistically significant associations between turbidity and GI illness were detected at lags 1, 2 and 7 days. PMID- 12745351 TI - The presence and antimicrobial susceptibilities of human-pathogen Vibrio spp. isolated from blue crab (Callinectes sapidus) in Belek tourism coast, Turkey. AB - Monitoring of Vibrio species by blue crabs (Callinectes sapidus) was carried out during the winter period in a selected area of the Belek, Antalya Gulf. Eighty three blue crabs were examined for Vibrio species. V. alginolyticus (30.1%), V. fluvialis (10.8%), V. damsela (9.6%), V. harveyi (3.6%), V. metschnikovii (3.6%) and V. vulnificus (2.4%) were isolated. V. vulnificus was the highest concentration (5 x 10(8) Vibrio ml(-1)) although it was only 2.4% isolated from blue crabs. The strains of different vibrio species were highly susceptible to doxycycline, tetracycline and ciprofloxacin. PMID- 12745352 TI - Contemplating drug monitoring systems in the light of Australia's "heroin shortage". PMID- 12745354 TI - Patterns and correlates of substance use amongst juvenile detainees in New South Wales 1989-99. AB - In the decade 1989 - 99 there have been significant changes in the patterns of substance use in the Australian community. Juvenile offenders have been a sentinel population of these emerging trends. The social and personal costs associated with adolescent substance use, especially where it leads to increased criminal offending requires urgent attention. This study was a replication of the 1989 and 1994 surveys of young people in detention in New South Wales, Australia. The 300 voluntary participants from nine detention centres had a similar demographic profile to participants of the previous surveys. They were predominantly male (90%) with a mean age of 16.5 years and an over-representation of Aboriginal and Torres Straits Islander peoples. The patterns of lifetime alcohol and tobacco use were stable over the decade, with particular increases in amphetamine, opioid and cocaine use since 1994. The more concerning pattern of at least weekly substance use revealed significant increases in cannabis, opioid and cocaine use since 1994, but a significant decrease in the frequent use of alcohol. This study also reports on high levels of negative health and psychosocial consequences of substance use, including overdose, among this group. High levels of self-reported depression and suicidal behaviours, family and gender issues are also discussed. Encouragingly, there was a relatively high level of self-recognized treatment need for substance use and mental health problems among the sample. This highlights further the growing need for the development and dissemination of novel interventions that harness this willingness and actively engage, motivate and maintain these young people in accessible, appropriate and effective interventions. PMID- 12745355 TI - The relationship of conduct disorder to attempted suicide and drug use history among methadone maintenance patients. AB - In order to examine the effects of a diagnosis of childhood conduct disorder (CD) on history of attempted suicide and drug use, unconfounded by early onset heroin use, 181 methadone maintenance patients who commenced heroin use after the age of 15 were interviewed. CD was diagnosed in 54% of patients. Compared to other patients, CD patients were younger and less educated. The onset of drug use, injecting drug use and heroin use occurred, on average, 2 years earlier than in other patients, and they had broader histories of injecting polydrug use. CD patients were more likely to have attempted suicide and to have been hospitalized after an attempt, and to have attempted suicide while enrolled in their current treatment. The current study indicates that a history of CD increases the risk of attempted suicide over and above the higher risks associated with injecting drug use per se. PMID- 12745357 TI - Similarities in outcomes for men and women after drug misuse treatment: results from the National Treatment Outcome Research Study (NTORS). AB - The National Treatment Outcome Research Study (NTORS) is a prospective, multi site treatment outcome study of drug misusers in the UK. This paper reports the characteristics and problems of male and female clients at intake to treatment, and changes in substance use, health problems, and criminal behaviour at follow up. The sample comprised 753 subjects (552 men and 201 women) followed-up 1 year after starting treatment in 54 programmes chosen to be representative of the main national treatment modalities. Men and women presented to treatment with different problems and characteristics. Women reported more frequent cocaine use, greater health problems, and were more likely to have a drug-using partner and be responsible for children. Despite different profiles of problems, men and women both made significant reductions in their problem behaviours following treatment. At 1-year follow-up, men and women reported reductions in drug use, health problems and criminal behaviour. However, women did not reduce their alcohol consumption significantly, and improvements in crime were less pronounced than for men. After controlling for pretreatment differences, gender was not predictive of any of the outcome measures reported. PMID- 12745353 TI - Patterns of co-morbidity between alcohol use and other substance use in the Australian population. AB - The present study describes patterns of co-morbidity between alcohol use and other substance use problems in the Australian population using data from the 1997 National Survey of Mental Health and Well-Being. Multiple regression analyses examined whether the observed associations between alcohol and other drug use disorders were explained by other variables, including demographic characteristics and neuroticism. We also assessed whether the presence of co morbid substance use disorders affected treatment seeking for a mental health problem. Alcohol use was related strongly to the use of other substances. Those who did not report alcohol use within the past 12 months were less likely to report using tobacco, cannabis, sedatives, stimulants or opiates. Higher rates again were observed among those with alcohol use disorders: half (51%) of those who were alcohol-dependent were regular tobacco smokers, one-third had used cannabis (32%); 15% reported other drug use; 15% met criteria for a cannabis use disorder and 7% met criteria for another drug use disorder. These associations were not accounted for by the demographic and other variables considered here. Co morbid substance use disorders (sedatives, stimulants or opioids) predicted a high likelihood of seeking treatment for a mental health problem among alcohol dependent people. PMID- 12745356 TI - The impact of methadone treatment on registered convictions and arrests in HIV positive and HIV-negative men and women with one or more treatment periods. AB - This study investigates criminality among 331 opiate abusers admitted to Stockholm's methadone maintenance programme (SMMP) between 1988 and 1992, and a comparison group of 1483 untreated opiate abusers. Information on arrests, criminal convictions, and intravenous drug abuse was obtained from official records. For both genders the annual rate of convictions decreased from 2.2 convictions per year during the 4 years prior to the first treatment, to 0.5 convictions during treatment, compared to 2.0 convictions for the comparison group. There was an even greater decrease in the rate of arrests for patients on methadone treatment. The decline was observed for both genders and in both HIV positive and HIV-negative patients. Rates of convictions among patients who had more than one treatment period were clearly reduced during each treatment period, and while the rate increased after they were expelled from treatment it remained at a lower level than during the 4 years prior to treatment. Thus, the methadone treatment is shown to have a profound positive effect on arrests and convictions, not only for patients remaining in treatment but also for those patients who were expelled from treatment involuntarily. PMID- 12745359 TI - Family functioning, alcohol expectancies and alcohol-related problems in a remote aboriginal Australian community: a preliminary psychometric validation study. AB - While a large proportion of Aboriginal Australians do not consume alcohol, those who do frequently show severe alcohol problems, and alcohol problems are associated with family conflict and violence. The aims were to examine the internal coherence and reliability of measures of family and alcohol measures for indigenous Australians in remote communities. This study involved 99 indigenous people from a remote North Queensland community. Questions were drawn from established questionnaires and administered verbally. Principle components factor analysis revealed three coherent and reliable measures of family conflict, independence, and cohesion. Expectancies of affective change had sound internal reliability for drinkers, and covaried with alcohol problems in the expected direction. The Alcohol Use Disorders Identification Test had two distinct factors relating to consumption and problems, but quantity/frequency measures may have limited reliability due to the common practice among drinkers of sharing alcohol. These measures should facilitate the detection and initial assessment of alcohol and family problems, and may be useful for evaluating change in future interventions that target family and alcohol problems. PMID- 12745360 TI - The monitoring of drug trends in Australia. AB - Recently, there has been increased recognition of the importance of drug information systems (DIS), highlighting the need for an internationally coordinated approach to data collection and advocating the regular assessment of a range of areas. Accurate information provides policy makers with the evidence to evaluate current strategies and to plan future strategies. An effective drug information system (DIS) must collect comprehensive, detailed and in-depth data, while also being sensitive to emergent trends and placing these changes into the context of longer-term trends. An integrated and comprehensive system combines both sensitive (or lead) and slower but more reliable lag indicators. This article reviews conceptual frameworks for DIS and developments in international systems. It then considers the range of DIS in Australia and then describes two integrated monitoring systems with an early warning function: the Illicit Drug Reporting System (IDRS) and the Drug Use Monitoring Australia (DUMA) Programme. Both systems collate sensitive lead indicators, and provide timely information about emerging drug trends in Australia. Together, these two systems are best placed to provide effective early warning of new trends in illicit drug markets, and constitute an important component of the overall approach to the monitoring of drug use and associated harms in Australia. PMID- 12745361 TI - Smoking intervention within alcohol and other drug treatment services: a selective review with suggestions for practical management. AB - This selective review was undertaken in order to highlight the need for alcohol and other drug treatment services to provide intervention for tobacco smoking to their clients. The reasons for the failure of treatment services to date to deal with nicotine addiction within their programmes are discussed and positive suggestions for change are proferred. In addition to the transformation of institutional culture which will be required, managers and staff of alcohol and other drug agencies need to know how best to implement smoking intervention within the treatment setting. The paper concludes with some practical suggestions for the management of intervention for tobacco smoking within treatment settings. These suggestions include: making decisions and formulating policies and procedures with regard to how tobacco smoking will be addressed; considering the particular physical, psychological and social/environmental factors that apply to substance abuse clients; building intervention around a simple structure such as the '5 A's'; encouraging and facilitating the use of nicotine replacement therapies; and allowing flexibility to tailor intervention to the individual. A great deal of further research is required to inform us as to how to intervene most effectively for tobacco smoking among this population group. PMID- 12745358 TI - Enough! or too much. What is 'excessive' kava use in Arnhem Land? AB - The objective of the study was to describe parameters for use in monitoring health, social and economic effects of kava use in Arnhem Land Aboriginal communities in the Northern Territory (NT). Kava has been used mainly in eight communities (population > 200), and in smaller associated homeland areas since 1982 with a total population of approximately 6800, using cross-sectional description and comparison using data from three kava-using communities. Interview data combined with health worker assessments were compiled using: (1) a sample (n=136) aged 16-34 years in one community in 2001-02; (2) a sample (n=101) aged 16 years and over in 2000 where physical assessments and biochemical and haematological data were also collected; (3) participant-observation in one community (133 people aged 18 years and over) during 1989-91. Kava, supplied illegally, was still being used in Arnhem Land in 2001-02. In 2000 dermopathy characteristic of heavy use, abnormally low body mass index (BMI), low blood lymphocytes and abnormally high gamma-glutamyl transferase (GGT) occurred more frequently with increased kava use. These acute effects emerge at average consumption levels of from 310-440 g/week of kava powder. When kava users in one community began to consume it at an average of 240-425 g/week from mid-1990, 19% of available cash resources were spent on kava with 11% of cash resources leaving the local community economy. The proportion of men drinking kava reached 70% and women 62% from mid-1990, with 20% of the population spending unprecedented amounts of time (14 + hours/week) in activities where kava was consumed. These parameters may be useful to monitor kava's adverse health, social and economic effects. Their association with increased kava use suggests that approaches to minimizing harm from its abuse may begin fruitfully with controlling supply. PMID- 12745362 TI - Harm reduction and alternative development in the Golden Triangle. AB - The authors of this Digest are anthropologists from Macquarie University, Sydney Australia. At the invitation of the German aid agency GTZ, they have been monitoring opium use and the impact of drug rehabilitation in Muang Sing Laos over the past 3 years. Their role is to provide analyses of how development projects alter the social make-up of their target communities and contribute to ways in which substance use/abuse is understood, practiced and controlled or reconfigured. In their consideration of development projects they take the perspective that harm reduction can and should include pre-emptive concern with factors that promote damaging drug use in the first place and furthermore, that these factors are at times the products of the distinct drug reduction strategies themselves. PMID- 12745363 TI - Evidence-based practice. PMID- 12745365 TI - The Houghton strain of Eimeria tenella: a review of the type strain selected for genome sequencing. AB - A landmark decision was taken in 2002 by the Biotechnology and Biological Sciences Research Council (BBSRC) in the UK to fund the genome sequencing of an eimerian (apicomplexan) parasite from the fowl. The project is a joint collaboration between the Institute for Animal Health (IAH) at Compton, UK and the Wellcome Trust Sanger Institute, Cambridge, UK (see http://www.sanger.ac.uk/Projects/E_tenella/ and http://www.iah.bbsrc.ac.uk/eimeria/). The species chosen, Eimeria tenella, is the best known member of the genus and the Houghton (H) strain is one of the most widely studied. The H strain was isolated in 1949 in the UK. It was maintained initially at the Houghton Poultry Research Station (HPRS), Houghton, and, following closure of that laboratory in 1992, thereafter at the IAH in Compton. At various times the parasite has been provided to other institutions and research groups carrying out coccidiosis research in the fowl. The H strain has been utilised in many fundamental studies on the eimerian life cycle and its relationship with the host. It has also been used to investigate the nature of drug resistance, and to derive attenuated lines-one of which, a "precocious line", is a component of a multivalent live attenuated coccidiosis vaccine (Paracox vaccine, Schering-Plough Animal Health). In this article, some immunological, epidemiological, genetic, and chemotherapeutic investigations with the H strain are reviewed, a summary is provided of the biological characteristics of the parasite and some of the core methods used to prepare purified extracellular life cycle stages for experimental studies or passage are given. PMID- 12745364 TI - Monocytosis is associated with the onset of leukocyte and viral infiltration of the brain in chickens infected with the very virulent Marek's disease virus strain C12/130. AB - Marek's disease (MDV) virus is mainly known for the induction of visceral lymphomas and lymphoid infiltration of peripheral nerves. Recently, additional tropism for the central nervous system has been recognised as a distinct feature of disease induced by very virulent MDV isolates. During the analysis of changes in the peripheral blood leukocyte subpopulations in chickens infected with either a virulent (HPRS-16) or a very virulent (C12/130) strain of MDV, we observed a marked monocytosis in chickens infected with C12/130. Perivascular cuffing in brain and mononuclear cell infiltration into the meninges of chickens infected with C12/130 were associated with the appearance of the monocytosis from 6-10 days post-infection. Our results show that a peripheral blood monocytosis may be a contributory factor in establishing or accelerating the severity of mononuclear infiltration into the meninges and perivascular spaces in the brain during infection by very virulent C12/130 strain of MDV. PMID- 12745366 TI - Reovirus identified as cause of disease in young geese. AB - The pathology, epizootiology and aetiology of a specific disease of young geese, which has been seen in Hungary for more than three decades, were investigated. The disease was characterised by splenitis and hepatitis with miliary necrotic foci during the acute phase, and epicarditis, arthritis and tenosynovitis during the subacute/chronic phase. Clinical signs usually appeared at 2 to 3 weeks of age and persisted for 3 to 6 weeks. From different organs of the affected birds, a reovirus was isolated in embryonated eggs and tissue cultures of Muscovy duck or goose origin, as well as in Vero cells. In experimental infections, the dominant features of the disease were reproduced in day-old and young goslings. The biological and partial molecular characterisation of one of the isolated strains (D15/99) showed that it was related to the reovirus described as the cause of a similar disease of Muscovy ducks. An RT-PCR method suitable for the detection of reoviruses was also elaborated and tested. This is the first report on the involvement of reovirus in arthritis of geese. PMID- 12745367 TI - Intravenous treatment with liposome-encapsulated dichloromethylene bisphosphonate (Cl2MBP) suppresses nitric oxide production and reduces genetic resistance to Marek's disease. AB - In this study the functional effectiveness of in vivo macrophage depletion using liposome-encapsulated dichloromethylene bisphosphonate (Cl(2)MBP) was examined in the chicken. The main target organs for systemic liposome-encapsulated Cl(2)MBP treatment are the spleen and the liver. Intravenous treatment with Cl(2)MBP of B(21)/B(21) chickens, genetically resistant to Marek's disease (MD), before challenge with the very virulent strain RB-1B, increased viral load in the blood and spleen after the first week and up to 6 weeks post-infection. In addition, Cl(2)MBP treatment dramatically increased tumour incidence and tumour load, especially in the spleens and livers of sick animals, but without affecting MD specific mortality of B(21)/B(21) chickens infected with RB-1B at 12 days of age. Nitric oxide (NO) is an important effector of the macrophage and has antiviral and antitumoural properties. NO has been shown to be one of the mechanisms triggered in resistance to Marek's disease. Intravenous treatment with Cl(2)MBP before infection with RB-1B induced a long-lasting decrease in numbers of macrophages and reduction in splenic inducible NO production associated with an absence of nitrate induction in the serum (up to 6 weeks p.i.). These results do not identify macrophage and NO production as major effector components in genetic resistance to Marek's disease, but underline their roles in limiting viraemia and tumour development in organs such as the spleen and the liver. PMID- 12745368 TI - Pathology and frequency of Cheilospirura hamulosa (Nematoda, Acuarioidea) in Galliformes hosts from backyard flocks. AB - This investigation reports data on frequency and pathology related to the nematode Cheilospirura hamulosa in 28 ring-necked pheasants (Phasianus colchicus) and 30 domestic chickens (Gallus g. domesticus) from backyard flocks of five localities of the State of Rio de Janeiro, Brazil. The prevalence of C. hamulosa in pheasants was of 14.3%, with a mean intensity of 1.5 and range of infection of 1-2. In chickens these values were of 26.7%, 4 and 1-12, respectively. Parasitized birds did not present with clinical signs. In spite of the low parasite burdens, the lesions induced by C. hamulosa were severe in the two species of investigated hosts, causing severe gross lesions in the gizzard such as haemorrhages, ulcers and thickening of the mucosa and cuticle, and single yellowish nodules on the caudoventral muscle. The microscopic lesions, also severe, were characterized by chronic diffuse inflammatory processes and ulcers in the mucosa, and granulomas in the muscular, submucosa and serosa layers of this organ. PMID- 12745369 TI - Occurrence of genotypes IV, V, VI and VIIa in Newcastle disease outbreaks in Germany between 1939 and 1995. AB - Forty-five velogenic Newcastle disease virus strains isolated in Germany between 1939 and 1995 were analysed by restriction enzyme digestion and sequencing to shed light on the relationships of past epizootics. Viruses derived from the period prior to 1970 belonged to a clade (IVea) of genotype IV comprising the earliest isolates from Europe, and could be isolated until the late seventies from poultry. Essex'70-like viruses, the prototype of genotype V, were already present at the beginning of the 1970-74 epizootic and in sporadic cases thereafter, indicating that these Newcastle disease outbreaks started in Western Europe. A genotype VI (subtype VIc) isolate was obtained in the early 1980s from a single outbreak in poultry. Outbreaks between 1993-95 were again part of a Western European epizootic caused by a genotype VIIa virus that was prevalent in the Far East. PMID- 12745370 TI - Nucleotide sequences of goose circovirus isolated in Taiwan. AB - We report the complete nucleotide (nt) sequences of eleven goose circovirus (GoCV) isolated in Taiwan. Nine out of the eleven isolates had a genome size of 1821 nt, whereas the remaining two isolates have a size of 1820 nt. Sequence comparisons of the eleven Taiwanese GoCV isolates and a German isolate revealed that these viruses could be divided into three distinct genetic groups. Group I contains the German isolate, group II contains three Taiwanese isolates, and group III contains eight Taiwanese isolates. Nucleotide differences between viruses of different genetic groups ranged from 7.0-7.7%, whereas the differences within the same group were only 0.2-1.0%. The most diversified sequences were found at a region between nt 27-72 of the viral genome, which corresponded to the right one-third of the 5' intergenic region. Open reading frame analysis shows that the genome of all Taiwanese GoCV isolates could encode four proteins: V1 (Rep, 293 amino acids), V2 (37 amino acids), C1 (capsid, 250 amino acids), and C2 (99 amino acids). The sizes of V1, C1 and C2 proteins of all Taiwanese isolates and the German GoCV isolates were identical. However, the size of V2 protein (37 amino acids), although identical in all Taiwanese isolates, was much smaller than that of the German isolate (120 amino acids). Moreover, the initiation codon of the V2 ORF of three Taiwanese isolates was ATA rather than ATG. Our result indicates that GoCV of multiple genetic groups might have been circulating in Europe and Asia, and these viruses differ in their nucleotide sequences, sizes of the genome, and sizes of the V2 ORFs. PMID- 12745371 TI - Passage of Salmonella through the crop and gizzard of broiler chickens fed with fermented liquid feed. AB - In vivo experiments were conducted in order to investigate the passage and bacterial reduction of Salmonella in the crop and gizzard of chickens when fed two different feeds. The chickens were fed dry conventional feed and fermented liquid feed. The fermented feed contains a relatively high concentration of lactic and acetic acid and lactobacilli. One and three week old broiler chickens were necropsied at short intervals after inoculation with Salmonella Enteritidis. Counts of Salmonella from the crop, gizzard, duodenum, caecum and colon/rectum were obtained. This revealed a sharper decrease of Salmonella in the anterior parts of the gastro-intestinal tract in chickens fed with fermented feed than in chickens fed dry feed. It is therefore concluded that fermented feed improves the barrier formed by the crop and gizzard. The reduction of Salmonella is fully realised in the crop and gizzard. The lower intestinal compartment did not show a substantial effect on the reduction of Salmonella. The performed in vivo method appeared to be an appropriate way to study intervention strategies that aim to control Salmonella by improving the barrier function of the upper gastro intestinal tract. PMID- 12745373 TI - A survey of blood lead levels in Mute Swans Cygnus olor. AB - Following bans on the use of most lead angling weights, the incidence of lead poisoning cases in Mute Swans started to fall and the population started to increase. However, surveys of lead levels in blood of rescued swans continue to show that a high proportion of the birds are carrying levels in excess of 1.21 mol/l. Since rescued swans, although rescued for many different reasons, might be a biased sample, a survey was made of apparently healthy birds living in flocks in the summer. These too showed that a high percentage of the birds in most of the flocks sampled had blood lead levels in excess of 1.21 mol/l. No source of this lead has been identified other than lead fishing weights; these may be long lost leads, current, but illegally used weights or "dust-shot" which it is still legal to use. Although many of the birds sampled are carrying lead burdens that are probably not harmful, others are seriously affected. Except in the most serious cases, it is not possible to recognise birds with elevated lead levels without taking a blood sample for assay. PMID- 12745372 TI - Comparison of the immune responses against Salmonella enterica serovar Gallinarum infection between naked neck chickens and a commercial chicken line. AB - The immune responses of indigenous naked neck (NaNa and Nana) and normally feathered (nana) chickens against a Salmonella Gallinarum (SG) infection were evaluated and compared with those of a commercial line (B-380). Groups of 28-day old chickens (NaNa, Nana, nana, and B-380) were immunized orally and subcutaneously with 50 microg of SG antigens. Control non-immunized animals were inoculated with sterile saline solution. All chickens were challenged with 1 LD(50) of SG and mortality was recorded daily for 20 days. Antibodies to SG were measured in sera before immunization, before the challenge, 10 days after the challenge, and at sacrifice. Peripheral blood lymphocyte proliferation assays were performed using concanavalin A and SG antigens. Results showed that non immunized Nana chickens exhibited the best natural resistance to Salmonella infection, since only 30% of them died. In contrast, all control B-380 chickens died by the 13th day. Immunization with SG induced immunity in chickens of all genotypes. Indigenous naked neck and normally feathered chickens showed a higher survival rate when compared with B-380 chickens. Immunized Nana chickens showed the highest antibody titres (P<0.05) as well as the highest thymidine incorporation in peripheral blood lymphocytes stimulated with con A or SG antigens (P<0.05). The results show that Nana chickens are the most resistant to SG infection and the best responders to vaccination with SG antigens. PMID- 12745374 TI - Apparent eradication of Mycoplasma synoviae in broiler breeders subjected to intensive antibiotic treatment directed to control Escherichia coli. AB - A Mycoplasma synoviae (MS)-free flock of broiler breeders was housed for brooding and rearing on an MS endemic farm. PCR revealed that the flock became infected within nine weeks. At 22 weeks the flock was transferred to a clean and disinfected house on a previously depopulated farm. The birds were then subjected to three treatments with fluoroquinolones due to recurrent Escherichia coli peritonitis and from the 32 weeks of age they received 600 ppm of oxytetracycline hydrochloride continuously in the feed. Monitoring by PCR showed a decrease in MS positive birds after 34 weeks of age and MS may have been eradicated as judged by consistent negative results in PCR. We conclude that intensive antibiotic treatments supported by adequate biosecurity could clear MS from infected broiler breeders. PMID- 12745375 TI - Protection of chickens against highly lethal H5N1 and H7N1 avian influenza viruses with a recombinant fowlpox virus co-expressing H5 haemagglutinin and N1 neuraminidase genes. AB - Inactivated whole avian influenza virus (AIV) vaccine provides protection against homologous haemagglutinin (HA) subtype virus, but poor protection against a heterologous HA virus. Moreover, it induces chickens to produce antibodies to cross-reactive antigens, especially nucleoprotein, which is limits AIV serological surveillance. In this study, a recombinant fowlpox virus co expressing HA (H5 subtype) and NA (NI subtype)genes of AIV was evaluated for its ability to protect chickens against intramuscular challenge with a lethal dose of highly pathogenic (HP) AIV. Susceptible chickens were also vaccinated by wing-web puncture with the parent fowlpox vaccine virus. Following challenge 4 weeks later with HPAIV, all chickens vaccinated with recombinant virus were protected, while the chickens vaccinated with either the unaltered parent fowlpox vaccine virus or unvaccinated controls experienced 100% mortality following challenge. This protection was accompanied by the high levels of specific antibody to the respective components of the recombinant vaccine. The above results showed that rFPV-HA-NA could be a potential vaccine to replace current inactivated vaccines for preventing AI. PMID- 12745376 TI - Lead and lead toxicity in domestic and free living birds. AB - At present, domestic and wild fauna are being exposed to aspects and factors which are foreign to the habitat in which they live. One that stands out is the enormous amount and variety of chemical compounds which, in many cases, are highly complex and which are constantly being released into the atmosphere, mainly from agricultural and industrial activity. All these substances affect some species more than others, whether they be plants or animals, from the most insignificant micro-organism to the most evolved species, among them birds. Finally, another cause of mortality in many birds is plumbism, namely death caused by the ingestion of lead. Lead has been one of the main causes of poisoning in man since ancient times due to its use in many activities although it is only recently that this toxicity has been recognized. Moreover, the use of lead pellets for shooting has resulted in the release into the environment of millions of these over many years, with serious repercussions for many bird species populations, which have ingested them either directly or indirectly. Added to this use of lead in cynegetic activities is the fate of the lead weights (sinkers or ballast) used by rod fishers, which sink to the bottom or accumulate on the banks of rivers, lakes, lagoons or reservoirs. The problem arises when these pellets or weights are ingested by birds, mainly Anatidae, which mistake them for the small stones or grit they use to triturate food in their gizzards. Small particles of lead enter the digestive tract, start dissolving in the form of lead salts, are incorporated into the bloodstream and the rest of the body, accumulate in organs like the liver or kidneys, and cause physiological or behavioural changes. When certain concentrations of lead are reached, the birds then die. If lead-poisoned birds are consumed by carrions or predators, the latter also ingest the lead so that they may also be affected or die from plumbism since, being a heavy metal, its degradation and/or elimination is very difficult. There is, therefore, no doubt that millions of birds die annually worldwide from lead poisoning (in the U.S.A., around 3,000,000), this problem being most acute in marshland. The solutions could include the introduction of legislation regulating or banning shooting, in the use of non-toxic ammunition in marshes and protected areas, the substitution of lead pellets for other non-toxic ones, such as steel, bismuth, tungsten or other suitable metals, and to go on studying other possible alternatives to end such a dramatic situation for birds all over the world. PMID- 12745378 TI - Phenotypic and genotypic differentiation of Campylobacter spp. isolated from Austrian broiler farms: a comparison. AB - An identification scheme based on restriction fragment length polymorphism of polymerase chain reaction products (PCR-RFLP) was developed to differentiate isolates of the genera Campylobacter, Arcobacter and Helicobacter. Based on the 16S rRNA gene of these genera, PCR amplified a 1216-bp fragment. The amplicons were digested with the restriction enzymes RsaI and EcoRV. Additional differentiation was obtained using a PCR-assay based on the hippuricase gene. Genotyping was performed on several reference strains from the National Collection of Typing Culture (NCTC), London, and on 130 field isolates. In parallel, a phenotypic differentiation was performed, in order to compare the results. In 119 cases (91.5%) the results obtained from the genotypic characterization were concordant with those from phenotypic testing. Co infections with Campylobacter jejuni and Campylobacter coli in two samples and seven hippurate-negative C. jejuni-strains were identified by the genotypic method. Furthermore, PCR-RFLP assays identified an atypical isolate as Campylobacter fetus/hyointestinalis. PMID- 12745377 TI - The contribution of humoral immunity to the control of avian reoviral infection in chickens after vaccination with live reovirus vaccine (strain 2177) at an early age. AB - The presence of maternal immunity in broilers does not preclude the successful protective immunization with an attenuated live reovirus vaccine at 1-day-old. This was recently demonstrated in a reovirus challenge model based on reovirus isolation from different organs (van Loon et al., 2002). Discrimination between challenge and vaccine virus was made by specific monoclonal antibody. Notably, at the day of challenge the levels of circulating reovirus-specific antibodies proved to be minimal or undetectable. This suggested an antibody-independent vaccination-induced immunity under the chosen conditions. In the present study we assessed the contribution of specific humoral responses in the same experimental model, by evaluating the control of a virulent challenge infection in the complete absence of B cells as a result of cyclophosphamide treatment. These experiments were performed in both SPF white leghorn chickens and commercially available broilers with passively acquired maternal antibodies. Our data demonstrate that the virus is controlled in the absence of actively produced antibodies, and is independent of B lymphocytes. This suggests that cellular immunity is sufficient for protection of broilers with maternal antibodies against reovirus infection following early age vaccination with live reoviral vaccine. PMID- 12745379 TI - Haemagglutinating activity of the lentogenic Newcastle disease virus strain MET95. AB - Using the rapid glass plate method, the Newcastle disease virus strain MET95 showed much weaker haemagglutination (HA) activity for chicken erythrocytes than 69 other Newcastle disease viruses, including 56 field strains isolated from chickens reared in Japan between 1988 and 2001. Using erythrocytes from other avian species, only the MET95 strain failed to show HA activity for erythrocytes from ducks, geese or pigeons. The haemagglutinin-neuraminidase protein of the MET95 strain was shown to have unique substitutions of isoleucine for thereonine and leucine at amino acide residues 216 and 552. It is suggested that these two substitutions might relate to the unique HA activity of the MET95 strain. This HA activity may be a useful marker for this strain. PMID- 12745381 TI - Hepatitis associated with Clostridium difficile in an ostrich chick. AB - A live 19-day-old male ostrich chick was euthanized and necropsied. It was one of 12 chicks in a group in which 8 had died with history of anorexia, diarrhoea and weight loss. The birds had been treated with amikacin, piperacillin and enrofloxacin. Necropsy of the ostrich revealed dehydration, mild ascites and serous atrophy of fat around the heart. The liver had numerous yellow tan foci on the capsular surface as well as on the cut surface. Caecal contents were watery. Microscopic examination of the liver revealed multifocal necrosis of hepatocytes with infiltration of heterophils mixed with fibrin, few lymphocytes, and multinucleated giant cells. A Gram stain of the liver revealed a few gram positive bacilli scattered within the necrotic foci. Clostridium difficile was isolated from the liver, and toxin A was detected by ELISA. A retrospective examination of approximately 1000 ostriches submitted during a seven year period to the laboratory system revealed seven cases of hepatitis due to Clostridium perfringens, two additional cases due to C. difficile and two cases due to C. sordelli. PMID- 12745380 TI - Development of a DIVA (Differentiating Infected from Vaccinated Animals) strategy using a vaccine containing a heterologous neuraminidase for the control of avian influenza. AB - The present paper reports of the development and validation of a control strategy for avian influenza infections in poultry. The "DIVA" (Differentiating Infected from Vaccinated Animals) strategy is based on the use of an inactivated oil emulsion vaccine containing the same haemagglutinin (H) subtype as the challenge virus, but a different neuraminidase (N). The possibility of using the heterologous N subtype, to differentiate between vaccinated and naturally infected birds, was investigated through the development of an "ad hoc" serological test based on the detection of specific anti-N1 antibodies. This was achieved using a baculovirus expressing a recombinant N1 protein. The A/ck/Pakistan/H7N3 virus was used as a vaccine and birds were challenged with the HPAI A/ty/Italy/4580/V99/H7N1 strain. The homologous H group ensured a clinical protection of 93% regardless of the vaccination scheme used, and was able to prevent viraemia and muscle colonization in the clinically healthy challenged birds. However, it was not able to prevent viral shedding. The "ad hoc" serological assay was developed as an indirect immunofluorescence test, and was validated using 608 field sera, and showed an "almost perfect agreement" (Kappa value) with the HI test, with relative sensitivity and specificity values of 98.1 and 95.7, respectively. The results of the present investigation suggest that the "DIVA" control strategy may represent a tool for the control of avian influenza infections in poultry. PMID- 12745382 TI - Characterization of Newcastle disease virus isolates obtained from Eurasian collared doves (Streptopelia decaocto) in Italy. AB - Eurasian collared doves (Streptopelia decaocto) are thought to originate from India and they have colonized, throughout the centuries, the Middle East and, more recently, Mediterranean countries such as Italy and Spain. In the present paper we report of the isolation and characterization of Newcastle disease viruses (NDV) obtained from Eurasian collared doves during 2000-2001, and compare them to isolates obtained from feral pigeons (Columba livia) during the same period. All isolates could be classified as avian paramyxovirus type 1 (APMV1) and belonged to the pigeon variant group (PPMV1), as their haemagglutinating activity was inhibited by mAb 161/617 which is specific for PPMV1. The intracerebral pathogenicity indices ranged from 0.68 to 1.38 and all isolates contained multiple basic amino acids at the deduced cleavage site of the fusion protein, which is a typical feature of virulent viruses. Phylogenetic analysis of the isolates indicate that 18/20 of these form a separate cluster from the isolates obtained from pigeons in the same period. These findings suggest that different lineages are circulating in feral pigeon populations, and that a separate lineage affects Eurasian collared doves. PMID- 12745383 TI - Expression of type X collagen, Indian hedgehog and parathyroid hormone related protein in normal and tibial dyschondroplastic chick growth plates. AB - Tibial dyschondroplasia (TD) is a form of aberrant endochondral ossification in chickens, in that a plug of avascular cartilage (TD lesion) is formed within the growth plate. Histologically, the lesion is filled with apparently transitional chondrocytes that have been unable to differentiate to hypertrophic chondrocytes. We have examined the spatial expression of mRNAs for type X collagen, Indian hedgehog (Ihh) and Parathyroid Hormone-related protein (PTHrP) in the TD growth plate by in situ hybridization in order to ascertain at which stage chondrocyte differentiation is arrested in TD. In the normal growth plate, type X collagen mRNA was expressed by both prehypertrophic and hypertrophic chondrocytes. Indian Hedgehog mRNA was detected in a band of prehypertrophic chondrocytes and PTHrP expression was localized to a narrow band of prehypertrophic chondrocytes and in osteoblasts within the diaphysis. In TD sections, collagen X expression was seen within differentiating cells, within a small number of lesion cells, and within hypertrophic chondrocytes on the diaphyseal side of the lesion. Ihh expression was also seen within the differentiating cells and throughout the lesion. These data indicate that chondrocyte differentiation is arrested at the transitional stage just prior to hypertrophy. Contrary to the previously reported PTHrP expression patterns in TD chicks by immunohistochemistry, PTHrP mRNA was not detected in the TD lesion. This observation probably reflects the cessation of PTHrP gene expression by chondrocytes in the more severe TD lesions. The results from the present study also imply that the arrest of cell differentiation in TD is independent of PTHrP and that endochondral ossification in the post-hatch avian growth plate may involve additional regulatory pathways. PMID- 12745384 TI - Virulence of six heterogeneous-origin Newcastle disease virus isolates before and after sequential passages in domestic chickens. AB - Four serial passages of six Newcastle disease virus (NDV) isolates were performed in two-week-old White Leghorns. The viruses were recovered from chickens (Ckn Live Bird Market and Ckn-Australia isolates), exotic (Yellow Nape [YN] Parrot, Pheasant, and Dove isolates) and wild birds (Anhinga isolate). Infected chickens were monitored clinically and humanely killed to sample tissues for histopathology and immunohistochemistry. Pathogenicity tests, to assess the virulence of the isolates for chickens, and sequence analysis of the fusion protein cleavage site were performed before and after passages. The moderately virulent Dove isolate became highly virulent with serial passage. The originally highly virulent Pheasant isolate had an increase in the intracerebral pathogenicity index (ICPI) and the intravenous pathogenicity index (IVPI) with passages in chickens. Virulence increase was not observed with Ckn-LBM, YN Parrot, Ckn-Australia, or Anhinga isolates after four chicken passages. The results demonstrate the high risk for domestic chickens represented by some NDV infected non-poultry species, such as doves. PMID- 12745385 TI - Functional comparison of heterophils isolated from commercial broiler chickens. AB - Heterophils from two pure lines (A and B) of commercial broiler chickens were isolated on days 1, 4, and 7 post-hatch to evaluate their ability to (1) phagocytose Salmonella enteritidis (SE) (2) degranulate when exposed to immune IgG opsonized SE, and (3) produce an oxidative burst. On days 1 and 4, heterophils from line A were functionally more efficient compared to heterophils from line B (p<0.05). By 7 days post hatch, heterophil functions for both lines were comparable. To further study the inheritance of heterophil functional efficiency, F1 reciprocal crosses (line C=male Bxfemale A; line D=male Axfemale B) were evaluated for functional activity and compared with the immunologically efficient (A) and non-efficient (B) parent lines. Heterophils from line D had a more efficient heterophil function (p<0.05) when compared to heterophils from C. These results suggest that heterophil function and efficiency can be genetically transferred to progeny. Moreover they indicate that heterophil function is sex associated and genetically controlled by the rooster since progeny of line A males maintained immunologically efficient characteristics whereas heterophils from the progeny of line B roosters remained immunologically inefficient. To our knowledge, this is the first report to describe a functional relationship between pure and F1 reciprocal crosses of broiler chickens with regard to heterophils and the innate immune response. PMID- 12745386 TI - Recruitment of rural community-dwelling older adults: barriers, challenges, and strategies. AB - The significant increase in the number of older adults in the USA, particularly in rural areas, has signaled the need for more research to address the health care needs of this complex, heterogeneous, and vulnerable population. However, accessing older adults in rural areas presents an especially difficult challenge for gerontological researchers. Barriers can include the normal physiological changes that accompany aging, prevalence of chronic illness and medication use, lower literacy rates, transportation issues, and a distrust of 'outsiders' and research in general. Specific strategies that may facilitate participation include the use of gatekeepers to gain entry, increased personal contact by researchers sensitive to the unique needs of older adults, and other media approaches tailored to the rural community. Understanding the characteristics of rural older adults, along with careful planning of recruitment strategies, is crucial to obtaining adequate participation and the acceptance of future research efforts. PMID- 12745387 TI - Racial bias in the assessment of cognitive functioning of older adults. AB - This study was undertaken to determine if the difference in assessed cognition between Black/African-American and White older adults was due differential item functioning (DIF) and/or differences in the effect of background variables. Participants were 15257 adults aged 50 and older surveyed in the Study of Asset and Health Dynamics of the Oldest Old (AHEAD) and Health and Retirement Study (HRS). The cognitive measure was a modified telephone interview for cognitive status. The analytic strategy was a multiple group structural equation model grounded in item response theory. Results suggest that most (89%) of the group difference could be attributed to measurement or structural differences, the remainder being not significantly different from zero (p=0.193). Most items displayed racial DIF, accounting for most of the group difference. After controlling for DIF, the group difference that remained could be attributed to heterogeneity in the effect of background variables. For example, low education was more deleterious for Black/African-Americans, and high income conferred an advantage only for Whites. These findings underscore the importance of efforts to generate culture-fair measurement devices. However, culture-fair assessments may attenuate, but not eliminate, group differences in assessed cognition due to the incommensurate action of background variables PMID- 12745388 TI - Caregiver role strain: the contribution of multiple roles and available resources in African-American women. AB - While the roles of spouse, parent, and employee are normative for middle-aged adults, strain associated with each role may increase when the role of caregiver is added. The objectives of the present study were to determine: (1). what is the relationship between the caregiver's other roles (marital, parental, employee), specific combinations of these roles, and role strain; and (2). what is the relationship between available resources (economic, social, and personal) and role strain? The study participants were 148 African-American females who provided care to elderly family members. A model using multiple linear regression was analyzed. Caregivers with higher depressive symptomatology were more strained. However, neither additional multiple roles, nor a combination of roles were significantly related to role strain. African-American caregivers experienced a wide range of caregiver role strain. It would be a disservice to African-American caregivers to operate under the assumption that they provide care with little emotional cost. Continued within group analyses are needed to understand differential outcomes of these caregivers. PMID- 12745389 TI - The dimensions of insight in people with dementia. AB - This study aims to characterize the factors determining the retention or loss of insight in dementia through: (i). a comparison of assessment procedures previously used to quantify loss of insight and (ii). a qualitative analysis of interviews with patients and carers. Semi-structured interviews were carried out with thirty-two people with dementia and their carers, which incorporated assessments via clinical interview, discrepancy ratings between patient and carer on an Activities of Daily Living scale, and prediction of performance on a memory task. The results of these were compared and supplemented with themes arising from qualitative analysis of the interviews. Significant differences were found between insight as measured by a prediction of performance paradigm and other methods of assessment. This may reflect a distinction between implicit and explicit knowledge. Analysis of interviews identified the following factors as contributing to people's awareness of their difficulties: (1). short-term frustration or distress; (2). continuous discontent; (3). lack of concern; (4). normalization of problems; (5). worry and anxiety; (6). defensiveness; (7). explicit denial; and (8). priority given to other problems. We conclude that insight in dementia is a complex, multi-dimensional, and value-laden concept. Emotional and behavioural aspects need to be considered as well as the cognitive, which is emphasized by traditional methods of assessment based on quantitative ratings. Implicit awareness may be accessible through subjects' predictions of performance on cognitive tasks and should be further investigated. An adequate account of insight in dementia should incorporate both objective and subjective measures in order to determine the interrelationships between organic changes, personality factors, and psychological mechanisms. A generally accepted, reliable protocol for the assessment of insight needs to be developed. PMID- 12745390 TI - Coping and defending: age differences in maturity of defence mechanisms and coping strategies. AB - Previous studies have examined either coping strategies or defence mechanisms; however, few have considered both. This research examined age differences in the type of defence mechanisms and coping strategies that people employ. In addition, gender differences, personality, and environmental variables were taken into account. The three age groups used in this study included: 17-23 year olds, 40-47 year olds, and 63-70 year olds. The youngest participants used significantly less mature defence mechanisms and significantly more immature defence mechanisms than the middle-aged and the oldest group. However, there was no significant difference in maturity of defence mechanisms between the middle-aged and the oldest group. In contrast, there were no age differences revealed for effectiveness of coping strategies people employ. One further interesting finding was that people with a higher purpose in life were more likely to use mature defence mechanisms. This research concludes that when developing theories on stress, psychologists might benefit from considering both defence mechanisms and coping strategies. PMID- 12745391 TI - Attitudes towards psychotherapy with older people among trainee clinical psychologists. AB - Therapists have been described as reluctant to work with older people. A legacy of pessimism towards psychotherapy with this group has been traced back to Freud's assertions that older people are no longer educable. It remains unclear to what extent these views continue to influence today's therapists. This study explores the attitudes of trainee clinical psychologists towards psychotherapy with this age group. A cross-sectional postal survey design was used and the data were analyzed using content analysis. Three hundred and seventy-one trainees responded, representing 38% of the total population of trainees in the UK. The majority of the trainees thought that work with older people provided the opportunity to apply psychological knowledge and skills, although therapists needed to respond differently when working with an older person. Factors to take into account included cognitive decline and physical health problems. The trainees described a number of rewards and challenges associated with working with older people. The responses suggest that although a significant minority of trainees continue to hold some negative stereotypes about therapeutic work with older people, positive attitudes are also clearly evident. Further research to explore the attitudes of other healthcare professionals is recommended and the implications for training and supervision are examined. PMID- 12745392 TI - Attitudes towards mental health care in younger and older adults: similarities and differences. AB - It is commonly assumed that older and younger adults have very different attitudes about seeking mental health services and that this is a major factor in reducing the use of mental health services by the elderly. However, little evidence exists to illustrate how elders actually perceive mental health care. Responses from a survey of 474 older adults age 65 and over were compared with data from a national survey of 1001 persons age 21-65. Results indicate similarities in many attitudes including likelihood of seeking treatment for severe mental disorders, importance of mental health care, and concerns about cost and coverage as barriers to care. Differences included use of services, perceptions about less severe disorders, referral sources, and preferred providers. Clinical, policy and public education implications are discussed. PMID- 12745393 TI - The use of sections 2 and 3 of the Mental Health Act (1983) with older people: a prospective study. AB - The use of the UK Mental Health Act (MHA) is under scrutiny with older people, especially in those with dementia and other organic mental disorder. Whilst research into use of the MHA with this group has been sparse, the small body of existing research suggests that the MHA is applied differently to older adults (i.e. those over 65 years). This multi-centre study identified all MHA assessments conducted over a prospective three-month period, and obtained detailed data on the circumstances behind assessment. The findings highlighted that older people assessed under the MHA tend to exhibit different behaviour patterns, circumstances and core characteristics to those under 65; older people were more likely to be detained because of self neglect and physical illness and also more often had a diagnosis of an organic mental disorder. Younger people were unlikely to have a diagnosis of organic mental disorder and were more likely to be judged as a risk to other people. Risk of suicide was particularly highlighted with the under 65 age group. Implications for legislative reform are discussed. PMID- 12745394 TI - Young AIDS migrants in Southern Africa: policy implications for empowering children. AB - Many AIDS-affected children in southern Africa engage in migration when household members fall sick or die from AIDS, or because they are sent to assist relatives. Despite this, little attention has been paid to the consequences of these movements for children's lives. Multi-method research, conducted in Lesotho and Malawi, revealed that children sent to live with kin commonly move over long distances and between urban and rural areas. They are generally not consulted or informed about these migrations and face a range of associated difficulties, particularly with integrating into new families and communities. Severed family ties exacerbate the difficulties faced by children who end up in institutions or on the streets. This paper advocates that policy approaches for those affected by AIDS should be children-centred and take into account the implications of migration at three levels. First, many of the difficulties children face could be overcome if they were familiar with the place and people they were moving to Second, children would be better able to cope with new situations if they were included in family discussions with decision-makers regarding their migration preferences. Third, maintaining ties with kin would ensure that children do not become distanced from their family and cultural heritage, which is essential for post-institutional support. PMID- 12745395 TI - Adherence issues in children and adolescents receiving highly active antiretroviral therapy. AB - The introduction of highly active antiretroviral therapies (HAART) for the treatment of paediatric HIV infection poses additional adherence challenges for children and families living with HIV A preliminary survey of 18 parents of children receiving HAART in Australia showed that although parents report high level of child adherence to HAART, specific features of the medication regimen, such as taste and number of medications made administration of HAART extremely difficult.Moreover, interaction between the treatment regimen and the day-to-day lives of families increases the adherence challenge. While some agreement exists in relation to the concerns families have about negative aspects of HAART; the diversity of issues suggests the need for ongoing and individualized support and information to families. PMID- 12745396 TI - Blood transfusion related paediatric HIV/AIDS in Ile-Ife, Nigeria. AB - This prospective study was aimed at determining the contribution of blood transfusion to paediatric HIV infection in Ile-lfe, Nigeria. It involved HIV screening of consecutive children presenting at the Obafemi Awolowo University Teaching Hospital (OAUTH), Ile-Ife, between March 1996 and March 2001, with any of the signs in the WHO clinical case definition for paediatric AIDS. The HIV serostatus of blood donors at the OAUTH was also extracted from the blood bank records. Of the 263 children who met the criteria for HIV screening, 35 (13.3%) were HIV-positive. Eighteen of the 263 children screened had a history of blood transfusion and 12 (66.7%) of the 18 were HIV-positive. Eleven (91.7%) of the 12 HIV-positive patients were transfused in private hospitals with blood collected from private laboratories. The blood with which the HIV-positive children were transfused was unscreened in three, screened in two and the HIV status unknown in the others. The sole voluntary donor was an HIV-positive father whose child received his unscreened blood. Only two (16. 7%) of the mothers of the previously transfused HIV-positive children were also HIV-positive. 'Paid'donors accounted for 94.3% of total donors in OAUTH blood bank records and cumulative HIV positivity was statistically significantly higher in 'paid' donors than in voluntary donors (p = 0.005). Wl conclude that transfusion with unsafe blood is an important route for HIV infection in symptomatic children and that HIV positivity is higher among paid donors. recommend the establishment of a national blood transfusion service, which is presently non-existent in Nigeria, and the enforcement of laws guiding blood transfusion. Voluntary blood donation should been encouraged and health workers in the private sectors educated on the link between blood transfusion and HIV infection. PMID- 12745397 TI - Patterns of condom use in urban males in Zimbabwe: evidence from 4600 sexual contacts. AB - The objective of this paper is to get detailed information on the dynamics of condom use with various partner types, including consistency of use, and to estimate the frequency of unprotected intercourse. The study is based on a prospective cohort study of 222 urban workers in urban Zimbabwe. Respondents' sexual behaviour and condom use were tracked for a period of six weeks using daily interviewer-administered questionnaires. In total, the observation period covers 9324 person-days, during which 4601 sexual contacts were reported. The results indicate that most sexual encounters with casual partners are protected, and condom use with these partners is very consistent. However, although the majority of sex acts with regular non-marital partners are protected, only four out of ten men with regular partners used condoms consistently. Because many men have both regular and casual partners, fewer than one in seven males used condoms in all their sex acts. We estimate that nearly half of all males have more than 85 unprotected sex acts per year. Although many of these acts are with spouses, who have comparatively low risk, we estimate that about one in five men have 25 or more unprotected acts with regular non-marital partners annually. WI conclude that regular partners are likely to be a major source of HIV transmission in Zimbabwe, partly because condom use is inconsistent and partly because they account for a large share of all sex acts. Given that over one in four adults in the general population are HIV-positive, and that some studies show that married persons have the highest HIV prevalence, it is important for future HIV programmes to focus on increasing levels of protection with regular partners. PMID- 12745398 TI - Serostatus disclosure, sexual communication and safer sex in HIV-positive men. AB - This study assessed HIV-positive men's sexual behaviours with partners at risk for infection, and examined the extent to which safer sex was associated with interpersonal communication variables, namely, (1). disclosure of one's seropositive status and (2). specific communication with partners about safer-sex practices. A total of 105 HIV-positive men (43% homosexual, 38% bisexual, 19% heterosexual), randomly sampled at an HIV outpatient clinic in Los Angeles, completed a behavioural questionnaire assessing events in their most recent sexual encounter with an HIV-negative or unknown serostatus partner. Results indicated that men who disclosed their seropositive status and explicitly discussed the topic of safer sex with their at-risk partners had a significantly higher prevalence of protected anal or vaginal intercourse than did men who disclosed only. The findings suggest that post-test counselling regarding the importance of disclosing one's seropositive status to sex partners should be augmented by behavioural interventions that enhance seropositive persons' skills in communicating explicitly with partners about safer sex to help reduce transmission of HIV. PMID- 12745400 TI - Intentional unsafe sex (barebacking) among HIV-positive gay men who seek sexual partners on the internet. AB - While unsafe sex has been reported throughout the HIV epidemic, the underlying assumption has been that most persons do not seek to purposely ham unprotected sex. Within the gay community, the term 'barebacking' has emerged to refer to intentional unsafe anal sex. The prevalence of barebacking is evidenced among gay men, particularly those who are HIV-positive, by the number of internet sites devoted to barebacking and the number of men seeking sexual partners through the use of the internet. To gain insight into barebacking, a sample of 112 HIV positive gay men were recruited from internet sites where men seek to meet each other for sex. The major it of participants (84%)reported engaging in barebacking in the past three months, and 43% of the men reported recent bareback sex with a partner of unknown serostatus. These results indicate the potential for widespread transmission of HIV to uninfected men by the partners they meet on the internet. Analyses revealed that men who reported bareback sex only with HIV positive partners scored lower in sexual adventurism than those who had bareback sex regardless of partner serostatus. A significant correlation was observed between defining masculinity as sexual prowess and intentional unprotected anal sex. There are serious implications for HIV prevention efforts, in that internet based education should be a priority in order to reach men who rely on this mechanism to find sexual partners. PMID- 12745399 TI - Concerns and coping with HIV: comparisons across groups. AB - To develop scales measuring major concerns about being HIV-positive and how people would cope with diagnosis, items were selected from the Ways of Coping Scale (Folkman et al., 1986)and generated by county health department HIV counsellors. Psychometric scale development involved two diverse samples of HIV test clients. Study I (health department, N = 272) yielded five Concerns with HIV scales and nine Coping with HIV scales. Factor structures did not differ between gay/bisexual men, heterosexual women or heterosexual men. In Study II (private non-profit gay-identified community HIV clinic, N = 227), LISREL confirmatory factor analyses cross-validated the Study I factor structures with no notable differences found. Some mean differences between genders and by sexual orientation were explained by different numbers of HIV-positive people known. Concern and Coping with HIV Scales (CCHIVS) are discussed for HIV/AIDS research and clinical use. PMID- 12745401 TI - Exposure to the 'SIDA dans la Cite' AIDS prevention television series in Cote' d'Ivoire, sexual risk behaviour and condom use. AB - This study assesses factors associated with viewing of 'SIDA dans la Cite', a weekly television soap opera on AIDS in Cote d'Ivoire, and the relationship between 'SIDA dans la Cite' viewing, sexual risk behaviour and condom use. The study uses across-sectional survey of 2150 respondents aged 15-49 in three regions. Logistic regression analysis was used to assess the effect of'SIDA dans la Cite' exposure on condom use. The results show that 65% of the sample had seen at least one 'SIDA dans la Cite' episode. Among viewers, 27% of males and 41% of females had seen ten or more episodes. Persons who had risky sexual partners were particularly likely to watch the programme. Women who had seen ten or more episodes were 1.4 times more likely than non-viewers to have used a condom in last sex. Men who had seen ten or more episodes were 2. 7 times more likely to have used a condom. We conclude that television soap operas on AIDS, such as 'SIDA dans la Cite',can be an important tool for promoting condom use. The programme was most appealing to viewers who engaged in risky behaviour, who are the core transmitters of the virus. HIV prevention programmes that provide continuous information, through multiple media channels or through series of broadcasts, are likely to have the greatest impact on condom use. PMID- 12745403 TI - The reliability and validity of the Modified Condom Outcome Expectancy Scale (MCOES) among an international sample of HIV-negative partners of people living with HIV/AIDS. AB - Safer sexual behaviour is vital in curbing the HlV epidemic in many developing countries where the epidemic is primarily transmitted through heterosexual sex. Reliable and valid assessment of factors related to HIV risk behaviours are important in testing behavioural theories as well as informing effective AIDS prevention programmes. The current study tests the reliability and validity of a modified version of the Condom Outcome Expectancy Scale (COES) among a sample of HI'-negative sexual partners (n = 100) of HIV-infected individuals in Pune, India, Chang Mai,Thailand and Kampala, Uganda. Internal consistency reliability was measured using Cronbach's alpha coefficient, which had a value of 0.80 for the entire sample, with site-specific values of. 0.78 for India; 0.75 for Thailand; and 0.79 for Uganda. Test-retest reliability was conducted to test the scale's stability over time with 60% of the sample, resulting in a Cronbach's alpha coefficient of 0.70. The scale's structure was explored by analyzing response scores on the items using principal components analysis, which yielded a two-factor solution. The study indicates the utility of a modified version of the widely tested COES across international settings. Such research is necessary in understanding intervention targets across international settings. PMID- 12745402 TI - Initial steps to developing the World Health Organization's Quality of Life Instrument (WHOQOL) module for international assessment in HIV/AIDS. AB - This paper reports on the three initial steps taken to develop the World Health Organization's Quality of Life instrument (WHOQOL) module for assessment of persons living with HIV/AIDS (PLWHA). First, a consultation of international experts was convened to review the suitability of the generic WHOQOL-100 for assessment of PLWHA. The experts proposed additional facets that are specific to the lives of PLWHA. Second, 42 focus groups (N = 235) were conducted by six culturally diverse centres-comprising of PLWHA, informal carers and health professionals-to (1). review the adequacy of the WHOQOL for PLWHA, (2). review the additional facets proposed by the experts, and (3). write additional facets and items for a pilot instrument. Third, results of steps 1 and 2 were consolidated, and a total of 115 items, covering 25 new facets and sub-facets for assessment of QoL specific to PLWHA, were prepared for pilot testing. The new facets included symptoms of HIV, body image, sexual activities, work, social inclusion, disclosure, death and dying, and forgiveness. The implications of cross-cultural QoL assessment for PLWHA are discussed PMID- 12745404 TI - Novel low-risk commercial sex practices in the parks of Vietnam. AB - This is a paper about a novel form of commercial sex practice recorded in Vietnam, and its implications for HIV/AIDS prevention strategies. Female commercial sex workers occupy urban parks in the evenings, offering on-site masturbation services to clients at cheap prices even by local standards. All sex acts are performed in public by workers who sit on benches, path edges or stools,often behind bicycles or open umbrellas in the quest for some semblance of privacy. Clients are local men, with only an occasional foreigner involved Many sex workers were involved, some having 15 to 20 or more clients a night. Sex workers and clients in Vietnam, a s elsewhere, operate at high risk of HIV/AIDS infection. As a service which minimizes risks, this form of commercial sex practice should perhaps be given serious consideration by policy makers and authorities for formal deployment in the fight against AIDS. PMID- 12745410 TI - Allergic and asthmatic reactions to alcoholic drinks. AB - Alcoholic drinks are capable of triggering a wide range of allergic and allergic like responses, including rhinitis, itching, facial swelling, headache, cough and asthma. Limited epidemiological data suggests that many individuals are affected and that sensitivities occur to a variety of drinks, including wine, beer and spirits. In surveys of asthmatics, over 40% reported the triggering of allergic or allergic-like symptoms following alcoholic drink consumption and 30 - 35% reported worsening of their asthma. Sensitivity to ethanol itself can play a role in triggering adverse responses, particularly in Asians, which is due mainly to a reduced capacity to metabolize acetaldehyde. In Caucasians, specific non-alcohol components are the main cause of sensitivities to alcoholic drinks. Allergic sensitivities to specific components of beer, spirits and distilled liquors have been described. Wine is clearly the most commonly reported trigger for adverse responses. Sensitivities to wine appear to be due mainly to pharmacological intolerances to specific components, such as biogenic amines and the sulphite additives. Histamine in wine has been associated with the triggering of a wide spectrum of adverse symptoms, including sneezing, rhinitis, itching, flushing, headache and asthma. The sulphite additives in wine have been associated with triggering asthmatic responses. Clinical studies have confirmed sensitivities to the sulphites in wine in limited numbers of individuals, but the extent to which the sulphites contribute to wine sensitivity overall is not clear. The aetiology of wine-induced asthmatic responses may be complex and may involve several co factors. PMID- 12745411 TI - Treatment of nicotine dependence with bupropion SR: review of its efficacy, safety and pharmacological profile. AB - Bupropion hydrochloride, an atypical antidepressant, is the first non-nicotine product that, in its sustained release form (bupropion SR), has been licensed as an aid for smoking cessation. The specialized literature on bupropion SR and smoking cessation is critically reviewed. The pharmacological profile, dosage and administration, contraindications, as well as the clinical efficacy, safety and tolerability data of bupropion are discussed. Recent reports suggest that its mode of actions might be different to what had originally been proposed. When prescribed appropriately, it appears to be a safe, well-tolerated and effective medication in combination with smoking cessation counselling--for a wide range of smokers, as shown in several multicentre double-blind, placebo-controlled clinical trials. Further research is needed on aspects such as the optimal duration of treatment, the potential role of combination therapy with NRT and psychological interventions, and to establish its effectiveness in smokers with other psychiatric disorders or when used with only minimal support by general practitioners. PMID- 12745412 TI - Neuroendocrine correlates of antisocial personality disorder in abstinent heroin dependent subjects. AB - The function of the central alpha-adrenergic, serotoninergic and dopaminergic systems was investigated in 30 heroin-dependent subjects, 6 - 8 weeks after detoxification and in 22 psychophysically healthy controls (group C). Twelve heroin-dependent subjects with antisocial personality disorder (ASPD) (group A), 18 heroin-dependent subjects without other Axis I and II pathologies (group B) were included among abstinent substance abusers. The norepinephrine (NE) function was evaluated by the GH responses to acute stimulation with clonidine (clon); the serotonin (5-HT) function by the PRL and cortisol (CORT) responses to acute stimulation with d-fenfluramine (d-fen) and the dopamine (DA) function was investigated by growth hormone (GH) and prolactin (PRL) responses to acute administration of bromocriptine (brom). Alpha-adrenergic sensitivity, as measured by the GH-clon test, was found significantly reduced in A subjects (ASPD), in comparison with B subjects and controls. PRL and CORT responses to d-fen were significantly blunted both in A and B subjects, in comparison with control subjects. DA receptors sensitivity seems to be reduced significantly in ASPD (A subjects); in contrast, heroin addicts without open psychiatric co-morbidity showed unimpaired responses to brom challenge; a significantly lower GH response to brom and a lack of PRL suppression in ASPD subjects could express D2 postsynaptic receptor hyposensitivity possibly related to DA gene variants associated to co-morbid disorder. In sum, the study of central monoamine function revealed an alteration of the 5-HT system in all detoxified heroin-dependent subjects. A significant reduction of alpha-adrenergic receptors sensitivity and the hyposensitivity of postsynaptic DA receptors in ASPD subjects suggest once again that specific biological correlates of psychiatric co-morbidity may characterize substance abusers subtypes. PMID- 12745413 TI - HIV treatment in drug abusers: impact of alcohol use. AB - Studies of alcohol use in HIV-1 infected patients have resulted in conflicting and limited information regarding prevalence, as well as impact on HIV replication, disease progression and response to antiretroviral therapy. Alcohol, drug abuse and past medical information, including antiretroviral treatment, were obtained using research questionnaires and medical chart review in 220 HIV-1 infected drug users. A physical examination was conducted and blood was drawn to evaluate immune measures and nutritional status. Heavy alcohol consumption, defined as daily or 3 - 4 times per/week, was reported in 63% of the cohort. Men (odds ratio (OR) = 2.6, 95% CI 1.13 - 5.99, p = 0.013), and participants between 35 and 45 years of age were three times more likely to be heavy alcohol users (p = 0.006 and 0.0009, respectively). Low serum albumin levels were more evident in heavy alcohol users than non-drinkers (p = 0.003). Heavy alcohol users receiving antiretroviral therapy were twice as likely to have CD4 counts below 500 than light or non-drinkers (95% CI, 1 - 5.5, p = 0.03), and highly active antiretroviral therapy (HAART)-treated heavy alcohol users were four times less likely to achieve a positive virological response (95% CI, 1.2 - 17, p = 0.04). Alcohol consumption is prevalent in our HIV-1 infected drug user cohort and significantly impacts both immunological and virological response to HAART treatment. PMID- 12745414 TI - Impact of tobacco use on the development of opportunistic respiratory infections in HIV seropositive patients on antiretroviral therapy. AB - The increased risk of developing lung diseases in cigarette smokers has been well recognized. The association between smoking and the risk of developing pulmonary infections in HIV-1-infected patients, however, which has not been established, was evaluated in the present study. Twenty-seven cases with lower respiratory infections (15 Pneumocystis carinii pneumonia (PCP), 12 TB cases) were compared with 27 age, gender, socio-economic and HIV status-matched patients, without history of respiratory diseases. Medical history and physical examinations were obtained every 6 months. Blood was drawn for CD4 and viral load measurements. A substantial number of HIV + smokers who developed PCP (one-third) had been on highly active retroviral therapy (HAART) for more than 6 months and prophylaxis had been discontinued. Multivariate analyses indicated that in HIV-infected people, after controlling for HIV status and antiretrovirals, cigarette smoking doubled the risk for developing PCP (p = 0.01). Multivariate analyses demonstrated that long-term smoking also increased the risk (2 x) of developing tuberculosis (p = 0.04). Moreover, daily tobacco use seemed to attenuate by 40% the immune and virological response to antiretroviral therapies. These findings indicate that tobacco use significantly increases the risk of pulmonary diseases in HIV infected subjects and has a potential deleterious impact on antiretroviral treatment. PMID- 12745415 TI - User views on supervised methadone consumption. AB - To assess the views of opiate-dependent individuals about supervised methadone consumption. Three groups of opinions were sought: (i). new patients referred for assessment and treatment, using rating scales; (ii). the consensus view of the Methadone Alliance (a national users' forum); and (iii). the consensus view of a local service users' forum. All three groups expressed the view that supervised consumption has an important place in methadone treatments. Users understand the need for daily supervision of methadone and are generally willing to accept it. Users' views provide support for the introduction of flexible methadone prescribing regimes incorporating supervised consumption. Privacy in pharmacies and the possibility of moving away from supervision are important elements in an acceptable programme. Supervised consumption is an important component of safe, effective and responsible methadone prescribing. PMID- 12745416 TI - Major disruptions of sleep during treatment of the opiate withdrawal syndrome: differences between methadone and lofexidine detoxification treatments. AB - Sleep disturbance experienced during methadone or lofexidine opiate detoxification was investigated in 118 opiate-dependent patients receiving inpatient detoxification treatment. Sleep was assessed at four time-points during opiate detoxification using a self-report questionnaire. Maximum sleep disruption occurred at completion of detoxification and during the protracted withdrawal period, with patients in the methadone group reporting higher levels of withdrawal symptoms, lower overall sleep, longer sleep latencies and significantly longer periods of time awake than lofexidine patients. Regression analyses demonstrated a significant relationship between sleep disturbance, protracted withdrawal and retention in treatment, in addition to the major treatment benefit of reduced sleep disturbance conferred by lofexidine treatment. PMID- 12745417 TI - Using saccadic eye movements as objective measures of tolerance in methadone dependent individuals during the hydromorphone challenge test. AB - Substitute methadone prescribing is one of the main modes of treatment for opiate dependence. This study examined the relationship between methadone dose (measured by daily dose and methadone's active (R)-enantiomer blood levels) and opiate receptor function. Nine subjects on substitute methadone (30 - 90 mg daily) received three subcutaneous injections 1.5 hours apart (saline, 5 mg and 10 mg hydromorphone, a short-acting opiate agonist) followed by measures of functional response in particular saccadic eye movements (SEMs), as well as self-report measures. Ten mg of hydromorphone significantly slowed SEM parameters (peak velocity by 15%, p < 0.005; peak acceleration by 20%, p < 0.025; peak deceleration by 26%, p < 0.025) and the SEM velocity changes correlated significantly with (R)-methadone levels (r = 0.844, p < 0.005) and with the oral dose of methadone being taken (r = 0.829, p < 0.005). Although a similar trend was observed for 5 mg, this was not significant. These finding suggest that, at higher methadone doses (resulting in higher plasma concentrations), there is significant tolerance to the action of agonists. Such studies may help in refining our understanding of the actions of methadone and the SEM measure could help in defining the degree of tolerance in individuals using street heroin. PMID- 12745418 TI - Relationship between drug use and prefrontal-associated traits. AB - The prefrontal cortex plays an important role in the strategic and emotional regulation of behavior. Both cognitive and neuroimaging studies have implicated prefrontal cortex in processes of reward and addiction. Prefrontal-associated neurobehavioral traits may be measured psychometrically with the Frontal Systems Behavior Scale (FrSBe), so it was hypothesized that self-ratings on this instrument would correlate with parameters of psychoactive drug use in a community sample. Modest but significant correlations were found for various parameters of tobacco use, even after controlling for demographic variables. Significant differences were noted in the mean scores when non-users were compared with users of cannabis, major stimulants (e.g. cocaine, amphetamine), and dissociative hallucinogens (e.g. phencyclidine, ketamine) and polysubstance use, particularly with the Disinhibition subscale. Smokers rated greater dysfunction than non-smokers on all three subscales, with ex-smokers showing intermediate ratings between the two. Polysubstance users showed greater dysfunction on the Disinhibition subscale compared to non-polysubstance users. In summary, this study further supports a relationship between prefrontal dysfunction and drug use in normal individuals, convergently with other methodologies for studying addiction. PMID- 12745419 TI - Effects of acute alcohol withdrawal on memory performance in alcohol-dependent patients: a pilot study. AB - Studies on the neuropsychological performance in detoxified alcoholic patients often begin by acknowledging that there is a cognitive impairment to be found. Only little attention has been paid to date to the question as to how acute alcohol withdrawal might affect cognitive functions. Twenty-nine alcohol dependent inpatients, nine in moderate alcohol withdrawal, treated with carbamazepine (group 1), 10 in mild alcohol withdrawal without pharmacological treatment (group 2), 10 in mild alcohol withdrawal with carbamazepine treatment (group 3) and 31 healthy subjects as controls (group 4) underwent repeated investigations using memory tests. The tests were performed on the first, third, seventh and fourteenth days of withdrawal. Immediate free recall of a word-list was impaired in the three patient groups in comparison with the control group on the 1st day. Thereafter no significant differences could be revealed between patients and controls. In a word-list recognition test the memory functions were not impaired in group 1 and group 2 in comparison with the control subjects. However, patients in group 3 showed impairment in this recognition test in comparison with the healthy subjects on the first and third days. The present study suggests that acute alcohol withdrawal impairs memory functions, especially free recall. This should be considered in treatment interventions in the early days of withdrawal. PMID- 12745420 TI - Reactivity of IGF binding protein-3 isoforms towards concanavalin A in healthy adults and subjects with cirrhosis. AB - The capacity of the liver to synthesize insulin-like growth factors (IGFs) and their binding proteins (IGFBPs) may be compromised by alcohol. The characteristics of IGFBP-3 variants obtained from healthy individuals and patients with alcoholic cirrhosis (ALC) were compared. Concanavalin A (Con A) affinity electrophoresis and ligand blotting demonstrated that there was a gradual change in carbohydrate properties of putative IGFBP-3 with progression of ALC from stages A to C. As many as 12 ionic species of IGFBP-3 could be distinguished, corresponding probably to variously glycosylated and/or phosphorylated isoforms of the core protein. Three of them reacted significantly with the immobilized Con A, the pattern being altered in patients with ALC. Patients with ALC in stage B exhibited the presence of clearly differentiated IGFBP-3 variants less and more Con A reactive, suggesting this stage to be a turning point with the most intensive changes in the IGF - IGFBP system. Because the glycosylation pattern is tissue specific, pathological post-translational modifications found for one glycoprotein (IGFBP-3) are probably shared by others of the same tissue origin. This may affect their susceptibility to proteolysis and subsequently their function. PMID- 12745421 TI - Deaths from drug abuse in northwestern Spain, 1992 - 97. AB - A total of 338 cases of death from acute reactions to drugs in Galicia (NW Spain) from 1 June 1992 to 31 July 1997 were examined. The average annual mortality rate was close to 70 cases. Male victims (90%) prevailed over female ones, the average age at death being 28.8 years. Most of the victims were habitual users who died in their own homes (40%) or nearby (30%). Polydrug use was the most common pattern, the most frequently detected drugs being opioids (281 cases), followed by alcohol (128 cases), benzodiazepines (92 cases) and cocaine (75 cases). Although the intravenous route prevailed (91%), oral and inhalation consumption of the drugs were also significant--the latter has grown significantly in recent times in relation to opioids and other drugs. How accurate the certificate of death can be depends on how thorough the investigation at the crime scene, autopsy room and laboratory are, as well as appropriate knowledge of the individual's history. The coordinated action of different health care institutions and use of available resources are crucial with a view to obtaining such data. PMID- 12745422 TI - No change in apoptosis in skeletal muscle exposed acutely or chronically to alcohol. AB - The pathogenic mechanisms responsible for the deleterious changes in ethanol exposed skeletal muscle are unknown, although apoptosis may be a causal process. We therefore investigated the responses of skeletal muscle to acute or chronic ethanol exposure in male Wistar rats. In acute studies, rats were dosed with ethanol (75 mmol (3.46 g)/kg BW) and killed after either 2.5 or 6 hours. In chronic studies, rats were fed ethanol as 35% of total dietary energy for 6 weeks. Apoptosis was determined by either DNA fragmentation or TUNEL (terminal deoxynucleotidyl transferase mediated dUTP nick end labelling) assays. The results showed that apoptosis was not increased in the ethanol-exposed muscle in both acute and chronic studies compared to appropriate controls. PMID- 12745423 TI - A computer model for reshaping of cells in epithelia due to in-plane deformation and annealing. AB - Although cell reshaping is fundamental to the mechanics of epithelia, technical barriers have prevented the methods of mechanics from being used to investigate it. These barriers have recently been overcome by the cell-based finite element formulation of Chen and Brodland. Here, parameters to describe the fabric of an epithelium in terms of cell shape and orientation and cell edge density are defined. Then, rectangular "patches" of model epithelia having various initial fabric parameters are generated and are either allowed to anneal or are subjected to one of several patterns of in-plane deformation. The simulations show that cell reshaping lags the deformation history, that it is allayed by cell rearrangement and that it causes the epithelium as a whole to exhibit viscoelastic mechanical properties. Equations to describe changes in cell shape due to annealing and in-plane deformation are presented. PMID- 12745424 TI - A model of human muscle energy expenditure. AB - A model of muscle energy expenditure was developed for predicting thermal, as well as mechanical energy liberation during simulated muscle contractions. The model was designed to yield energy (heat and work) rate predictions appropriate for human skeletal muscle contracting at normal body temperature. The basic form of the present model is similar to many previous models of muscle energy expenditure, but parameter values were based almost entirely on mammalian muscle data, with preference given to human data where possible. Nonlinear phenomena associated with submaximal activation were also incorporated. The muscle energy model was evaluated at varying levels of complexity, ranging from simulated contractions of isolated muscle, to simulations of whole body locomotion. In all cases, acceptable agreement was found between simulated and experimental energy liberation. The present model should be useful in future studies of the energetics of human movement using forward dynamic computer simulation. PMID- 12745425 TI - Shear-slip Mesh Update Method: implementation and applications. AB - The Shear-slip Mesh Update Method (SSMUM) is being used in flow simulations involving large but regular displacements of one or more boundaries of the computational domain. We follow up the earlier discussion of the method with notes on practical implementation aspects. In order to establish a benchmark problem for this class of flow problems, we define and report results from a two dimensional viscous flow around a rotating stirrer in a square chamber. The application potential of the method is demonstrated in the context of biomedical design problem, as we perform an analysis of blood flow in a centrifugal left ventricular assist device, or blood pump, which involves a rotating impeller in a non-axisymmetric housing. PMID- 12745426 TI - Three-dimensional trabecular alignment model. AB - Trabecular alignment theory has been used to quantify Wolff's Law of bone remodeling. A three-dimensional finite element scheme was developed to analyze the bone remodeling phenomenon. The mathematical model proposed by Mullender et al. and later modified by Smith et al. was adopted to simulate the surface-based trabecular resorption and formation processes. Enhancements incorporated into the previous model include: mapping into three-dimensions, controlling the remodeling signal's passage through marrow, controlling the finite distance the signal may pass through the bone matrix, and including non-bone material in the finite element model. After the model is explained and thoroughly studied, three dimensional implant surface geometries are simulated. PMID- 12745428 TI - On the effects of cyclic transversal forces on osseointegrated dental implants: experimental and finite element shakedown analyses. AB - This paper is concerned with the mechanical strength of fixed osseointegrated dental implants subjected to cyclic external loads, applied mainly in a direction orthogonal to their axis. Such a loading condition, seen as a basic design action for the implant, has been given little attention so far. Experimental results and numerical simulations, performed on two- and three-dimensional Finite Element models, are discussed. The shakedown theory is used to show that a common implant design (threaded fixture-abutment-connection screw) is susceptible of low-cycle fatigue failure under loading conditions well within the working range, even if the same design is able to withstand loading of the same type, but applied monotonically, much in excess of the working values. The shakedown analyses give an indication of several possible failure modalities: the low-cycle fatigue either of the implant or of the connection screw, or the loosening of the connection screw itself. Experimental and numerical results are in good qualitative agreement, and both suggest that the issue of transversal cyclic loading on fixed dental implants should be carefully reconsidered in the design phase. PMID- 12745427 TI - Material changes in osteoporotic human cancellous bone following infiltration with acrylic bone cement for a vertebral cement augmentation. AB - Bone cement infiltration can be effective at mechanically augmenting osteoporotic vertebrae. While most published literature describes the gain in mechanical strength of augmented vertebrae, we report the first measurements of viscoelastic material changes of cancellous bone due to cement infiltration. We infiltrated cancellous core specimen harvested from osteoporotic cadaveric spines with acrylic bone cement. Bone specimen before and after cement infiltration were subjected to identical quasi-static and relaxation loading in confined and free compression. Testing data were fitted to a linear viscoelastic model of compressible material and the model parameters for cement, native cancellous bone, and cancellous bone infiltrated (composite) with cement were identified. The fitting demonstrated that the linear viscoelastic model presented in this paper accurately describes the mechanical behaviour of cement and bone, before and after infiltration. Although the composite specimen did not completely adopt the properties of bulk bone cement, the stiffening of cancellous bone due to cement infiltration is considerable. The composite was, for example, 8.5 times stiffer than native bone. The local stiffening of cancellous bone in patients may alter the load transfer of the augmented motion segment and may be the cause of subsequent fractures in the vertebrae adjacent to the ones infiltrated with cement. The material model and parameters in this paper, together with an adequate finite-element model, can be helpful to investigate the load shift, the mechanism for subsequent fractures, and filling patterns for ideal cement infiltration. PMID- 12745429 TI - The most appropriate position and number for absolute anchorages for orthodontic tooth movements. AB - Absolute anchorages proved to be very effective for orthodontic tooth movements. We used a 3D digitizer to record each tooth on pre-treatment diagnostic and post treatment predictive setup models and then 3D coordinate system conversion was performed to make the coordinate values comparable. An arithmetic calculation of vector and moment based on the orthodontic forces and the tooth displacement under preliminary premises undertaken to decide the most favorable position and number for absolute anchorages. Position--For two-dimensional and three dimensional calculations, the most appropriate positions for absolute anchorages should theoretically be on the line of resultant force (2D) and the plane (3D) where the total moment effect tends to be zero. Number--As for the number of the absolute anchorages needed, it depends on the number of target teeth. Different combinations of target teeth provide different sets of results. PMID- 12745430 TI - Skeletonization of volumetric angiograms for display. AB - The display of three-dimensional angiograms can benefit from the knowledge of quantitative shape features such as tangent and curvature of the centerline of vessels. These can be obtained from a curve-like skeleton representation. If connectivity and topology are preserved, and if geometrical constraints such as smoothness and centeredness are satisfied, it is possible to estimate length, orientation, curvature, and torsion. It is also required that no part of the original object be left unrepresented. An efficient method for the identification of such shape components is developed. First, a suitable representation is obtained using a voxel coding approach to yield connected and labeled unit-thick paths. The desired features are estimated from a smoothed version of the skeleton produced by a moving average filter. The computational cost is linear, of the order of N(object), the total number of object voxels contained in the binary volumetric data. The method is also shown to be robust to boundary noise. Examples are discussed. PMID- 12745431 TI - The influence of cavity shape on the stresses in composite dental restorations: a finite element study. AB - Dental restoration adhering to the cavity exhibits fundamentally different load transfer mechanisms from non-adhering restorations. It is therefore questionable that traditional cavity designs are optimal from a purely mechanical point of view when working with composite materials. Drawing from general engineering experience, it can be hypothesised that smooth, well rounded designs with bevelled margins are superior. A finite element model is used in the present investigation to determine the stress field in four different cavity designs as it develops during the curing of the restoration. The results show that a significant reduction of the stress along the adhesive interface between the tooth and the restoration can be achieved through the use of a rounded cavity shape. They also show that the adoption of bevelled margins leads to a reduction of the stress concentration at this location. These results are confirmed by a set of experimental results published in the literature. It is concluded that adhering restorations will perform better from a mechanical point of view if an appropriate cavity shape is selected. PMID- 12745432 TI - Constitutive laws and failure models for compact bones subjected to dynamic loading. AB - Many biological tissues, such as bones and ligaments, are fibrous. The geometrical structure of these tissues shows that they exhibit a similar hierarchy in their ultra- and macro-structures. The aim of this work is to develop a model to study the failure of fibrous structures subjected to dynamic loading. The important feature of this model is that it describes failure in terms of the loss of cohesion between fibres. We have developed a model based on the lamellar structure of compact bone with fibres oriented at 0, 45 and 90 degrees to the longitudinal axis of the bone and have studied the influence of the model parameters on the failure process. Bone porosity and joint stress force at failure were found to be the most significant parameters. Using least square resolution, we deduced a phenomenological model of the lamellar structure. Finally, experimental results were found to be comparable with our numerical model. PMID- 12745433 TI - Quantitative approach for the prediction of tooth movement during orthodontic treatment. AB - The orthodontic treatment is aimed to displace and/or rotate the teeth to obtain the functionally correct occlusion and the best aesthetics and consists in applying forces and/or couples to tooth crowns. The applied loads are generated by the elastic recovery of metallic wires linked to the tooth crowns by brackets. These loads generate a stress state into the periodontal ligament and hence, in the alveolar bone, causing the bone remodeling responsible for the tooth movement. The orthodontic appliance is usually designed on the basis of the clinical experience of the orthodontist. In this work, a quantitative approach for the prediction of the tooth movement is presented that has been developed as a first step to build up a computer tool to aid the orthodontist in designing the orthodontic appliance. The model calculates the tooth movement through time with respect to a fixed Cartesian frame located in the middle of the dental arch. The user interface panel has been designed to allow the orthodontist to manage the standard geometrical references and parameters usually adopted to design the treatment. Simulations of specific cases are reported for which the parameters of the model are selected in order to reproduce forecasts of tooth movement matching data published in experimental works. PMID- 12745434 TI - Three-dimensional simulation of fracture repair in the human tibia. AB - Finite element models of bones can be generated based on images obtained non invasively in the clinic. One area where such models may prove useful is in the assessment of fracture healing of long bones. To establish the feasibility of such a proposal, a three dimensional finite element model of a fractured tibia was generated, and a model of tissue differentiation and bone regeneration was used to simulate the progress of healing under two different loading magnitudes. Healing is successful under the lower load and unsuccessful under the higher load -this proves that the model has the potential to identify loads that would cause healing to fail. Following a proposal by Richardson et al. [J. Bone Jt Surg. Vol. 76B (1994) pp. 389-394] that the bending stiffness can be used to assess the extent of healing, the bending stiffness was computed during healing--it was shown that the stiffness changed in a similar manner that observed clinically. In conclusion, the paper establishes that 3D computer simulation could be a tool for assessment of the fracture healing under different orthopedic treatments. PMID- 12745436 TI - CD9-alpha6beta1 interactions in migratory parietal endoderm cells. AB - Tetraspanins modulate the function of a variety of membrane proteins, including integrin receptors. We show here that the tetraspanin CD9 preferentially coimmunoprecipitates with the alpha6beta1 integrin heterodimer in F9-derived parietal endoderm cells in comparison to F9 stem cells. We also show that CD9 function-blocking antibody inhibits parietal endoderm migration in an embryoid body outgrowth assay. In addition, both CD9 and alpha6beta1 colocalize with vinculin to apparent focal adhesion sites in parietal endoderm cells. The data presented here suggests a role for CD9 in localizing the integrin to the focal adhesion. In addition, the data suggest a role for CD9 in alpha6beta1 mediated migration of parietal endoderm. PMID- 12745435 TI - Cell adhesion induces the plasminogen activator inhibitor-1 gene expression through phosphatidylinositol 3-kinase/Akt activation in anchorage dependent cells. AB - Type-I plasminogen activator inhibitor (PAI-1) is the primary inhibitor of both tissue- and urokinase-type plasminogen activators (t-PA, u-PA) and is thus a primary regulator of plasminogen activation and possibly of extracellular proteolysis. In anchorage-dependent cells, the PAI-1 gene was regulated by cell adhesion. PAI-1 gene expression was induced more evidently in cells adhered to the culture plate than in nonadherent cells. In this study, we investigated the signal pathway of the PAI-1 gene expression regulated by cell adhesion. We found the induction of both PAI-1 mRNA and protein, when cells adhered to culture dish, was inhibited by the PI-3 kinase specific inhibitors (Ly294002 and wortmannin). The cells seeded on collagen-1 coated plate with low serum further demonstrated that the PAI-1 gene expression was prolonged by the cell adhesion. The above mentioned PI-3 kinase specific inhibitors also blocked the PAI-1 maintenance when cell adhered to collagen-1 coated plate. In addition, we found that both PI-3 kinase and its downstream molecule, Akt, were activated more evidently in adherent cells than in nonadherent cells. Furthermore, we transfected antisense oligodeoxynucleotides of Akt (AS-ODN-Akt) into cells to block the expression of Akt and found that the induction of PAI-1 mRNA was also inhibited. Hence, we conclude that the induction of PAI-1 gene expression is cell adhesion dependent and is through PI-3 kinase and Akt activation. PMID- 12745437 TI - Differential regulation of membrane bound and soluble ICAM 1 in human endothelium and blood mononuclear cells: effects of interferon beta-1a. AB - The membrane-associated Intercellular Adhesion Molecule 1 (mICAM 1) is fundamental for adhesion of leukocytes to endothelial cells. A soluble form of ICAM 1 (sICAM 1) exists in the human serum, and is seen as marker of disease activity in patients suffering from Multiple Sclerosis (MS). High levels of sICAM 1 have been detected in MS patients benefiting from interferon beta (IFNbeta) treatment, but little is known on the molecular origins of sICAM 1. This study investigated the interrelationship and the mechanisms of production of sICAM 1 and mICAM 1 in human endothelium (Human Umbilical Vein Endothelial Cells, HUVECs) and mononuclear leukocytes (MNL) upon stimulation with IFNbeta-1a and other inducers. We found that the expression of mICAM 1 and the release of sICAM 1 are differentially regulated in both these cytotypes. HUVECs and MNL express specific mRNA for both mICAM 1 and sICAM 1, and modification of the content of each of these transcripts results in regulation of both the ICAM 1 isoforms. We show that IFNbeta-1a is strong regulator of the ICAM 1 RNA splicing machinery. Effect of IFNbeta-1a over expression of the ICAM 1 isoforms might have a relevant immunomoregulatory role in Multiple Sclerosis. PMID- 12745438 TI - Src-/- fibroblasts are defective in their ability to disassemble focal adhesions in response to phorbol ester/hyaluronan treatment. AB - Exogenous hyaluronan promotes a rapid recruitment of Src to lamellae of mutant active H-ras transformed fibroblasts and an Src- and RHAMM (CD168)-dependent increase in random motility. These responses are accompanied by a loss of vinculin-positive lamellae focal adhesions. Nontransformed immortalized wild-type fibroblasts (WT) do not increase random motility in response to hyaluronan alone, but do increase motility in response to a combination of PMA treatment followed by hyaluronan. PMA treatment alone increases the number of lamellae/cell, percentage of cells with lamellae and number of focal adhesions/lamellae. Subsequent addition of hyaluronan does not affect the number of lamellae/cell but reduces both the number of focal adhesion/lamellae and the percentage of cells forming focal adhesion-positive lamellae. These effects are prevented by blocking RHAMM antibodies and mimicked by agonist RHAMM antibodies. Src-/- fibroblasts exhibit a limited response to PMA but do not increase motility or disassemble focal adhesions in response to a subsequent addition of HA. Rescue of Src-/- fibroblasts with either SrcA or c-Src restores response to close to WT levels. These results suggest that Src activity is uniquely required for both PMA and PMA induced hyaluronan regulation of random motility and focal adhesion turnover. PMID- 12745439 TI - Intra- and interactive touch on the face. AB - Experiments were performed in which the subjective size of steel balls was judged by the method of absolute magnitude estimation (AME). The balls were rolled actively ("scripting") by the right index finger pad of the subject over the passively receiving areas of the face, namely the forehead, cheek and the vermilion border of the lower lip. These areas were stimulated on the subject's own person (intra-active touch), on another person's face (interactive touch) and by another person upon the face of the subject (interactive touch). Results indicate that size perception appears to be governed primarily by the mechanical characteristics of the stimuli, including the balls and the sites on the face. When the subject's finger was used to script upon another's face, there were significant differences among or between the sites. When another person scripted upon the subject's face there were no significant differences among any of the sites. The results are discussed considering mode of stimulation and tissue mechanics. PMID- 12745440 TI - Cross-modal recognition of shape from hand to eyes in human newborns. AB - The hypothesis that the ability to coordinate information between tactual and visual modalities is present at birth and dependent on perceptual inherent structures was tested in human newborns. Using an intersensory paired-preference procedure, we showed that newborns can visually recognize the shape of an object that they have previously manipulated with their right hand, out of sight. This is an experimental evidence that newborns can extract shape information in a tactual format and transform it in a visual format before they have had the opportunity to learn from the pairings of visual and tactual experience. This is contrary to a host of theories and models of perceptual learning, both traditional (empiricist philosophers) and modern (connectionist). PMID- 12745441 TI - Thermosensory intensity and affect throughout the perceptible range. AB - The thermosensory system was evaluated psychophysically in 12 healthy volunteers, spanning the full range of tolerable temperatures. Subjects provided ratings of (1) perceived thermal intensity, (2) perceived pleasantness or unpleasantness, and (3) perceived pain intensity after placing either one hand or foot in a temperature controlled water bath. Of particular interest were the interrelationships among the three perceptual measures, and differences between heat and cold. The relationship between perceived intensity and (un)pleasantness was different for hot vs cold stimuli. Specifically, for a given perceived thermal intensity, cold stimuli were rated as less pleasant or more unpleasant than hot stimuli. Similarly, for a given pain intensity, cold stimuli were rated as more unpleasant than hot stimuli. As warm temperatures increased and as cold temperatures decreased, stimuli were perceived as being unpleasant before they were perceived as being painful. The difference in transition temperatures for unpleasantness vs pain for heat averaged 1.4 degrees C, while the same difference for cold averaged 5.6 degrees C. Thus, there was a fourfold difference in the range of unpleasant but non-painful cold vs hot temperatures. Pain intensity and unpleasantness ratings were significantly higher for heat stimuli applied to the foot vs hand. In contrast, there was no significant body site difference for pain intensity or unpleasantness ratings of cold stimuli. All of these results reveal important differences in the processing of cold vs hot stimuli. These differences could be exploited to differentiate processing relevant to discriminative vs affective components of somesthetic perception, in both the innocuous and noxious ranges. PMID- 12745442 TI - Force matching at the elbow joint is disturbed by muscle soreness. AB - These experiments are concerned with the ability of human subjects to match isometric torque in their elbow flexor muscles when biceps of one arm is made sore. Pain was induced by injection of hypertonic saline. Subjects were asked to generate a level of torque, 30% of maximum, with one arm, the reference arm. To achieve the required torque, subjects were given visual feedback. Subjects were then asked to match this torque with their other arm, the indicator arm. In control measurements, subjects were consistent in their matching ability and often were quite accurate. However, when biceps of one arm was made sore, subjects consistently and significantly underestimated the level of torque being generated by the sore arm. Painful heat applied to the skin over biceps produced a similar pattern of errors. Heating skin remote from elbow flexors had no significant effect. One interpretation of these findings is that the nociceptive input from the sore region of skin or muscle leads to reduced excitability of the motor cortex. That, in turn, disturbs the relationship between the centrally generated effort and motor output, leading to matching errors. PMID- 12745443 TI - The vibrations of texture. AB - The Pacinian channel has been implicated in the perception of fine textures (Hollins et al., Somatosens Mot Res 18: 253-262, 2001a). In the present study, we investigate candidate codes for Pacinian-mediated roughness perception. We use a Hall effect transducer to record the vibrations elicited in the skin when a set of textured surfaces is passively presented to the index finger. The peak frequency of the vibrations is found to decrease systematically as spatial period increases. The power of the vibrations--weighted according to the spectral sensitivity of the Pacinian system--increases with spatial period for all but the coarsest surfaces. By varying the scanning velocity, we manipulate the temporal and intensive characteristics of the texture-induced vibrations and assess the effect of the manipulation on perceived roughness. We find that doubling the scanning velocity does not result in the substantial decrease in roughness predicted by a frequency theory of vibrotactile roughness perception. On the other hand, the effects of speed on roughness match those of speed on power. We propose that the roughness of a fine surface (spatial period<200 microm) is a function of the Pacinian-weighted power of the vibrations it elicits. PMID- 12745444 TI - Time-dependence of SI RA neuron response to cutaneous flutter stimulation. AB - Spike discharge activity of RA-type SI cortical neurons was recorded extracellularly in anesthetized monkeys and cats. Multiple applications (trials) of 10-50 Hz sinusoidal vertical skin displacement stimulation ("flutter") were delivered to the receptive field (RF). Analysis revealed large and systematic temporal trends not only in SI RA neuron responsivity (measured as spikes/s and as spikes/stimulus cycle), but also in entrainment, and in phase angle of the entrained responses. In contrast to SI RA neurons, the response of RA skin afferents to comparable conditions of skin flutter stimulation exhibited little or no dynamics. The occurrence and form of the SI RA neuron response dynamics that accompany skin flutter stimulation are shown to depend on factors such as stimulus frequency and the locus of the recording site in the global cortical response pattern. Comparison of recordings obtained in near-radial vs tangential microelectrode penetrations further reveals that the SI RA neuron response dynamics that occur during skin flutter stimulation are relatively consistent within, but heterogeneous across column-sized regions. The observed SI RA neuron response dynamics are suggested to account, in part, for the improved capacity to discriminate stimulus frequency after an exposure ("adaptation") to skin flutter stimulation (Goble and Hollins, J Acoust Soc Am 96: 771-780, 1994). Parallels with recent proposals about the contributions to visual perception of short-term primary sensory cortical neuron dynamics and synchrony in multineuron spike activity patterns are identified and discussed. PMID- 12745445 TI - Sensory function above lesion level in spinal cord injury patients with and without pain. AB - Patients with spinal cord injury (SCI) may or may not develop central neuropathic pain despite having cord lesions of apparently the same site, extension and nature. The consequences of the cord lesion in the central nervous system and the mechanisms underlying pain are unclear. In this study, we examined sensory detection and pain thresholds above injury level in 17 SCI patients with central neuropathic pain, in 18 SCI patients without neuropathic pain, and in 20 control subjects without injury and pain. The SCI pain group had significantly higher cold and warm detection thresholds compared with the SCI pain free group and controls and higher tactile detection thresholds compared with the SCI pain free group. No difference in pain or pain tolerance thresholds was seen among pain and pain free SCI patients. These data suggest changes in somatosensory function in dermatomes rostral to the segmental injury level linked to the presence of central neuropathic pain in SCI patients. The results are discussed in relation to current concepts of pain inhibitory and facilitating systems. PMID- 12745446 TI - Glutamate receptor blockade alters the development of intracortical connections in rat barrel cortex. AB - We tested the hypothesis that glutamate receptor mediated activity is required for the postnatal development of intracortical connections in layers II/III of rodent barrel cortex. To block glutamate receptors, a slow release polymer (elvax) loaded with a glutamate receptor antagonist (D-AP5) was targeted subdurally over the future rat barrel cortex on P0 (day of birth). On P14-16 biotinylated dextran amine (BDA) was injected under the elvax into all layers to label neurons retrogradely. A BDA injection was made stereotactically at the mirror site of the untreated hemisphere of each animal. The animals survived to P22-24. Injection sites and retrogradely labeled cell bodies were identified in tangential sections in relation to the barrel map. D-AP5 treated and untreated hemispheres were matched according to the location of the injection site in the barrel map. Glutamate receptor blockade did not prevent the growth of intrinsic projections, but altered their organization. The normal row-like asymmetry of connections in untreated hemispheres was lacking in the D-AP5 treated cortex (ANOVA, p=0.02). Cortical activity mediated through glutamate receptors contributes to the correct development of connections between barrel columns in layers II/III. PMID- 12745447 TI - Barrels XV. Proceedings and abstracts of a 2-day meeting. Cocoa Beach, Florida, USA. October 31-November 1, 2002. AB - Approximately 80 colleagues attended this 2-day meeting from 31 October to 1 November 2002 in Cocoa Beach, FL. Summaries of the speakers' presentations are followed by abstracts of the 44 posters shown. PMID- 12745448 TI - Short-term exercise and platelet activity, sensitivity to agonist, and platelet leukocyte conjugate formation. AB - Strenuous exercise may be partially responsible for cardio-vascular events. The aim was to investigate the platelet activity, reactivity and different platelet leukocyte-conjugate formation following maximal short-term exercises. Fifteen healthy non-smokers underwent three isokinetic maximal tests on a SRM cycle ergometry system with durations of 15, 45 and 90 s. Blood samples were taken after a 30-min rest, immediately before and after exercise, and 15 min and 1 h after completion of exercise. Platelets were detected flow-cytometrically by CD41, and activated platelets by CD62P. In addition, stimulation of the platelets in vitro with 7.5 microM TRAP-6 was initiated. For testing platelet-leukocyte conjugates, antibodies against CD45, CD14 and CD41 were used. After the exercise tests the percent of non-stimulated CD62P-positive platelets (%PC) was unchanged. In contrast, an increase in %PC (CD62P) TRAP-6 stimulated (15-s test: 37.2+/-10.3 to 46.2+/-12.3%, P < 0.05; 90-s test: 40.6+/-9.5 to 51.7+/-10.2%, P < 0.01) and in platelet-granulocyte, platelet-lymphocyte, and platelet-monocyte conjugate formation 15 min after exercise (45- and 90-s test; P < 0.05) were observed in comparison with the changes on the control day. The changes nearly reversed 1 h after exercise. Maximal short-term exercise only leads to a moderate increase of platelet reactivity and to an increase in the different platelet-leukocyte conjugates. The implications of the changes in platelet-leukocyte conjugate formation should be investigated in future studies. PMID- 12745449 TI - Human platelets contain estrogen receptor alpha, caveolin-1 and estrogen receptor associated proteins. AB - Venous thrombosis is a risk factor for oral estrogen treatment in men and women. Platelet responses mediated through estrogen receptors may be involved in these processes. However, expression of estrogen receptors and estrogen receptor associated protein have not been defined clearly in human platelets. Therefore, the present study was designed to characterize estrogen receptors and estrogen receptor-associated proteins in human platelets. Platelets were obtained from normal female and male blood donors between the ages of 25 and 65 years. Estrogen receptors (ERalpha and ERbeta), heat shock proteins (hsp70 and hsp90), endothelial nitric oxide synthases (eNOS), caveolin-1, -2 and -3 and calmodulin were analyzed in total platelet lysate by immunoblotting. Expression of ERalpha and ERbeta, hsp70, hsp90, eNOS, calmodulin, and caveolin-1 were observed in both sexes. Expression of all proteins was similar in platelets from males and females. Results of the present study suggest for the first time that both estrogen receptors alpha and beta, caveolin-1 and other proteins associated with estrogen receptors and production of nitric oxide are present in platelets of both males and females. PMID- 12745450 TI - CD34 expression on platelets. AB - Flow cytometric analysis of human platelets from healthy volunteers by high resolution immunophenotyping accomplished with enzymatic amplification staining revealed the expression of CD34. Assessment of the platelets with five different monoclonal antibodies to the three different classes of CD34 epitopes suggested that platelets express a unique glycoform of the molecule. Expression of CD34 was verified by immunoblotting. Platelet activation resulted in a marked decrease in surface CD34. CD34 expression has significant implications for the function of platelets as well as for the determination of hematopoietic progenitor cells. PMID- 12745451 TI - Concurrent responses elicited by isolated activation of platelet Gq-coupled receptors, in vitro: a novel approach for their separation and analysis. AB - This study tested the hypothesis that aggregation mediated by activation of a single G(q)-coupled receptor can be studied quantitatively if four concurrent but distinct components of the observed platelet response, autocrine stimulation, shape change (SC), aggregation and deaggregation, are separately measured. Responses mediated by two G(q)-coupled receptors, the TXA(2) and the P2Y(1), were assayed by a novel, kinetics-based turbidimetric approach. Blocking the autocrine stimulation with a cocktail of receptor antagonists revealed rapid and sustained SC that largely masked the aggregation. Mathematical removal of the SC contribution from the changes in optical density indicated that selective activation of either the TXA(2) or the P2Y(1) receptor was sufficient to induce a small aggregation (DeltaOD = 0.1-0.2) that was reversed rapidly by a concurrent deaggregation. Model-derived rate constants of SC, aggregation and deaggregation described the observed complex time course of their concurrency well. Laser light scattering aggregometry confirmed the rapid deaggregation of platelet aggregates following TXA(2) or P2Y(1) receptor-mediated formation. Saturable concentration response curves of net aggregation were elicited at EC(50) values 6-15 higher than those of SC. The utility of this approach was further demonstrated by the separation of the four components of the response mediated by concurrent 5-HT(2A) and alpha(2A)-adrenoceptor activation, as well as the P2Y(1) and alpha(2A) adrenoceptor, and the autocrine stimulation and SC mediated by the 5-HT(2A) receptor. We propose that the novel approach described here is necessary to study the receptor pharmacology and the kinetics of concurrent platelet responses in vitro as demonstrated by platelet aggregation elicited by selective activation of the TXA(2) or the P2Y(1) receptors. PMID- 12745452 TI - A study of whole blood platelet and white cell aggregation using a laser flow aggregometer. AB - Both platelet aggregation and white blood cell aggregation are involved in pathological processes such as thrombosis, atherosclerosis and chronic inflammation. People in older age groups are likely to suffer from cardiovascular diseases and may have increased white cell and platelet aggregation which could contribute to this increased risk. This study aimed to compare white cell and platelet aggregation between different age and gender groups. Whole blood white cell aggregation and platelet aggregation were carried out on healthy volunteers using cytometric techniques. It was found that both white cell and platelet aggregation in the elderly group (white cell aggregation median value, 0.08; range, 0.02-0.14; platelet aggregation median value, 0.32; range, 0.1-0.39) were significantly higher (P = 0.017 for white cell aggregation, P = 0.007 for platelet aggregation) than in the younger group (white cell aggregation median value, 0.05; range, 0.01-0.14; platelet aggregation median value, 0.18; range, 0.07-0.36). No significant differences were found between the gender groups. PMID- 12745453 TI - Giant electron-dense chains, clusters and granules in megakaryocytes and platelets with normal dense bodies: an inherited thrombocytopenic disorder. AB - A woman and her male child were referred because of life-long thrombocytopenia, moderately increased platelet size, and absence of laboratory findings suggesting immune thrombocytopenia or defective platelet function. Evaluation of their platelets in the electron microscope revealed the presence of large organelles never seen before in human platelets. Examination of bone marrow from the mother and her son in an earlier study revealed that the giant platelet organelles originated in megakaryocytes. The present study has focused on the continuing development of the aberrant organelles in circulating platelets. The smallest subunits were electron-dense fragments and hollow-cored bodies observed in the dense tubular system (DTS). The dense fragments formed chains that became thicker, resulting in clusters, and clusters formed the large electron opaque bodies. Hollow-cored, almost hexagonal subunits also formed chains that interacted with each other to form target-like organelles. The multi-layered target organelles tended to become completely electron dense and difficult to distinguish from the opaque bodies. How two different types of aberrant organelles can develop in the same megakaryocyte/platelet system and both originate from channels of the DTS is unknown. Partial clarification stemming from analytical electron microscopy and ultrastructural cytochemistry will be presented in a subsequent report. PMID- 12745454 TI - Rapid and cost-effective identification of Candida species using multipoint inoculation of CHROMagar Candida media, cycloheximide sensitivity and carbohydrate assimilation tests. PMID- 12745455 TI - The biology and diagnostic applications of cadherins in neoplasia: a review. AB - Cadherins are a large family of calcium-dependent "cell adhesion molecules". In addition to their structural role, cadherins have a number of important functions in the control of cell growth and differentiation. This paper reviews the biology of cadherins and their role in neoplastic transformation, development of the invasive and metastatic phenotype, effects on tumour morphology and the applications of cadherin immunolabelling in tumour diagnosis. Potential areas of future investigation and diagnostic applications are discussed. PMID- 12745456 TI - Quantitative evaluation of AgNORs in bone tumours. AB - AIMS: Primary tumours of bone present a significant diagnostic and therapeutic challenge at times. Silver stained nucleolar organiser regions (AgNORs) have been widely used in a variety of tissues but with a limited study on bone tumours. Our study was aimed at the evaluation of AgNOR count in various neoplastic lesions of bone. METHODS: : One hundred biopsies of bone lesions were included in this study. Five samples of foetal lumbar vertebrae obtained from foetal autopsies were taken as control. The study included 58 males and 42 females with age ranging from 5 to 70 years. Fifty-two cases were malignant while 48 were benign in nature. Silver staining for nuclear organiser regions was performed according to one-step silver staining technique in these cases. NORs seen as black dots were counted in the nuclei of 100 cells. RESULTS: Our study revealed that the mean count was highest in malignant lesions (4.00+/-0.69) compared with benign lesions (2.16+/-0.43) and normal bone (1.32+/-0.14). Statistically, the AgNOR count showed a significant difference (P<0.001) in all these lesions. CONCLUSIONS: The results of the current study revealed that malignant lesions had a greater mean AgNOR count than benign tumours and the normal bone. Thus, quantification of AgNORs strongly correlates with the type as well as aggressiveness of the bone tumour and is diagnostically useful in tumour differentiation. PMID- 12745457 TI - A clinicopathological review of 34 cases of inflammatory breast disease showing an association between corynebacteria infection and granulomatous mastitis. AB - AIM: Granulomatous mastitis is a rare condition of unknown aetiology. The great majority of cases has not been associated with bacterial pathogens if women with mammary tuberculosis are excluded. We noted that some women in Auckland with a histological diagnosis of granulomatous mastitis had both microbiological and histological evidence of corynebacteria infection and aimed to study this further. METHODS: Thirty-four women were reviewed who presented with inflammatory breast disease and had microbiological specimens from which corynebacteria were isolated and/or histological specimens containing coryneform bacteria. These 34 cases were compared with 28 controls with similar histology but no evidence of corynebacteria infection. RESULTS: Twenty-seven (79%) of the cases and 21 (75%) of the controls had histological and/or cytological evidence of suppurative granulomas. Fourteen of the 34 cases also had Gram-positive bacilli (GPB), recognisable as coryneform bacteria, in histological sections. In all cases the bacilli were confined to empty spaces, consistent with dissolved lipid, and were surrounded by neutrophils and, frequently, suppurative granulomas. Corynebacterium species were isolated from 52 of 116 microbiological specimens taken from the 34 cases. Forty of these 52 cultures were pure. Twenty-four of the cultures were further classified biochemically and using 16S rRNA gene sequencing. Twenty of the 24 were lipophilic Corynebacterium species and 14 were identified as Corynebacterium kroppenstedtii. The cases were more likely to present with fever or neutrophilia and more often formed sinuses than the controls but other clinical features were similar. Maori and Pacific Islanders accounted for 77% of the women across both groups. CONCLUSION: We suggest granulomatous mastitis can be associated with corynebacteria infection, particularly infection by C. kroppenstedtii. The significance of this finding, which has previously been described in only a single case report, is discussed. PMID- 12745458 TI - Immunohistochemical characterisation of pulmonary hyaline membrane in various types of interstitial pneumonia. AB - AIMS: Hyaline membrane (HM) in diffuse alveolar damage (DAD) pattern is frequently detected in the acute stage of interstitial pneumonia (IP). To determine the exact nature of HM, we investigated immunohistochemically 25 cases of HM-containing IP. METHODS: The cases examined using various kinds of antibodies were four cases associated with rheumatoid arthritis, five with usual interstitial pneumonia, two with dermatomyositis, five with viral infection, one case with progressive systemic sclerosis and eight cases caused by other agents. RESULTS: HM mostly reacted with antibodies to PE10 (SP-A), Factor VIII, KL-6 and EMA and, interestingly, stained for AE1/AE3, CK19, and Hup-1 in some cases, but was negative for PTAH staining. However, the immunoreactivities of HM varied even within the same disease or section. CONCLUSIONS: The immunohistochemical heterogeneity of HM suggests that HM may be formed by different mechanisms in various types of IP. Our findings also suggest that the main components of HM are derived from alveolar epithelial cells and their proteins, some including cytoplasmic element of CK19, and also from serum factors, but not fibrin. The immunohistochemical characteristics of HM in DAD pattern will aid understanding of the significance of HM formation in IP. PMID- 12745459 TI - CD44s expression correlated with the International Neuroblastoma Pathology Classification (Shimada system) for neuroblastic tumours. AB - AIM: CD44 is a cell adhesion molecule that plays an important role in the cascade of metastasis and progression of human malignant tumours. A large family of variants or isoforms, generated by alternative splicing of a single gene, has been reported to be involved in the malignant process by conferring metastatic potential to non-metastatic cells. The objective of this study was to compare the expression of CD44 standard molecule with the International Neuroblastoma Pathology Classification (INPC) for neuroblastic tumours, a histological grading system based on the Shimada system for predicting the clinical outcome in neuroblastic tumours. METHODS: Paraffin blocks of primary neuroblastic tumour were graded according to the INPC system into favourable and unfavourable histological types. Tumour tissues were then analysed with immunostaining using monoclonal antibodies against the CD44 epitope. RESULTS: In this retrospective study, 32 cases of primary neuroblastic tumour were collected. Based on the INPC system, 13 cases (40.6%) had a favourable histology while 19 cases (59.4%) were of unfavourable histology. CD44 standard (CD44s) expression was detected in 69.2% of neuroblastic tumours with favourable histological type while 89.5% of tumours with unfavourable histological type did not express CD44s staining. Statistically, there was significant correlation between morphological grading of neuroblastic tumours based on the INPC and the presence of CD44 staining (Fisher's exact test, P<0.05). CONCLUSION: This study shows that there is a significant correlation between CD44s expression and tumour histology based on the INPC in which CD44s non-expression is correlated with an unfavourable histological type and the expression of CD44s with a favourable type. Therefore, the analysis of CD44s expression should be recommended as an additional biological marker in the initial routine staging of the disease. PMID- 12745460 TI - Adrenocortical tumours in children and adults: a study of pathological and proliferation features. AB - AIMS: To demonstrate the correlation between morphological and proliferation parameters with the biological behaviour of adrenocortical tumours (ACTs) in different age groups. METHODS: Seventy-six cases of ACT were evaluated comparing tumours of children (< or =15 years, n=24) and adults (>15 years, n=52) regarding morphological features and proliferation rates. These parameters were also compared in clinically benign (CB) and clinically malignant (CM) tumours. Tumours were analysed according to Weiss' histological parameters. Proliferation rates were assessed by MIB-1 labelling index and silver binding nucleolar organiser regions (AgNORs) were quantified by image analysis. RESULTS: ACTs in children presented higher proliferative activity than in adults. In adult's ACTs, MIB-1 labelling index (cut-off, 10%) (P=0.037) and the AgNOR pattern (P<0.001) had influence over the disease-free survival (DFS). The AgNOR pattern type I was associated with CB-ACT and the AgNOR pattern type II with CM-ACT. The AgNOR pattern type III was characteristic of children's ACTs and type IV was characteristic of non-neoplasic adrenal glands. CONCLUSION: ACTs in children had a different morphological spectrum and higher proliferative activity than those tumours in adults, which should be always analysed as a separate group. In adults, our findings reinforce that the morphological criteria proposed by Weiss are an efficient way of differentiating CB-ACT from CM-ACT. PMID- 12745461 TI - Epithelioid trophoblastic tumour: report of a case in the fallopian tube. AB - A case of epithelioid trophoblastic tumour (ETT), occurring in a fallopian tube of a 39-year-old woman, is reported. The patient presented with a positive pregnancy test, but continued to have 'periods'. A palpable right adnexal mass was noted that was confirmed on ultrasound. The mass was removed together with the uterus, omentum and associated ovary. Careful examination of the uterus revealed no evidence of either an antecedent tumour or intra-uterine pregnancy. Histologically, the tubal mass displayed sheets and islands of large, relatively uniform, mitotically active polyhedral cells, with surrounding necrosis. The immunoprofile of the tumour was atypical in that alpha-inhibin and epidermal growth factor were weakly positive, but other results were consistent with the diagnosis of ETT. The patient received a foreshortened course of standard EMACO (etoposide, actinomycin-D, methotrexate, vincristine, and cyclophosphamide) combination chemotherapy for high-risk gestational trophoblastic disease. Serum beta-hCG fell from a pre-operative level of 52 000 U/mL to non-pregnant levels within two courses and she remains well and disease-free 12 months post diagnosis. PMID- 12745463 TI - A discussion of cases in the 2001 RCPA-AQAP Chemical Pathology Case Report Comments Program. AB - AIM: We present a descriptive analysis of the 10 case reports distributed in the Royal College of Pathologists of Australasia (RCPA) and the Australasian Association of Clinical Biochemists (AACB) Chemical Pathology Patient Report Comments Program to assess the quality of interpretative commenting in clinical biochemistry in 2001. METHOD: Participants were asked to comment on a given set of biochemistry results attached with brief clinical details. All responses received were translated into key phrases and graphically presented on a histogram. An expert panel was asked to evaluate the appropriateness of the key phrases and to propose a suggested composite comment. RESULTS: While the majority of comments received were felt to be acceptable by the expert panel, some comments were felt to be inappropriate or misleading. As comments on laboratory reports may affect clinical management of patients, it is important that these comments reflect accepted practice and current guidelines. CONCLUSION: The Patient Report Comments Program may play an important role in continuing education and possibly in quality assurance of interpretative commenting. PMID- 12745462 TI - An autopsy case of gallbladder cancer developing in a Japanese man with cerebrotendinous xanthomatosis: genetic analysis of the sterol 27-hydroxylase and p53 genes. AB - Cerebrotendinous xanthomatosis (CTX) is a rare autosomal recessive lipid-storage disorder characterised by xanthomas, neurological dysfunctions and premature atherosclerosis. A case of a well differentiated adenocarcinoma of the gallbladder occurring in a 57-year-old Japanese man with CTX, confirmed clinically, biochemically and at autopsy is reported together with analyses of the sterol 27-hydroxylase (CYP27) and p53 genes. A missense mutation of the p53 (G for C) was detected in the gallbladder adenocarcinoma. Direct sequence analysis also showed a silent mutational substitution of unknown significance, C for A, in CYP27 at codon 89. In the past, CTX patients have only demonstrated this infrequently, indicating no direct relationship between CYP27 dysfunction and tumour development. Thus, the present case of gallbladder cancer appears to be a chance occurrence. PMID- 12745464 TI - Evaluation of a Western blot technique (Helicoblot 2.1) for the diagnosis of Helicobacter pylori infection in children. AB - AIM: We evaluated the performance of Helicoblot 2.1 which differentiates the reactivity to each of the various Helicobacter pylori antigens, and compared the results with those obtained by standard techniques (rapid urease test and histological examination of gastric biopsy) in symptomatic children of different ages living in Antalya, Turkey. METHODS: Eighty-eight children (mean age, 9.15 years) were divided into two groups. The first group included 66 children who were found to be infected with H. pylori. The second group included 22 children who were negative for H. pylori. Serum samples collected from all patients were tested for H. pylori IgG antibodies by immunoblot assay (Helicoblot 2.1). RESULTS: The sensitivity, specificity, positive and negative predictive values for detection of H. pylori infection were 80%, 100%, 100% and 85%, respectively. In children under 7 years of age, the sensitivity of the test was found to be lower than other age groups (P<0.05). No relationship was found between peptic ulcer and cagA antibody positivity (P>0.05). CONCLUSION: Helicoblot 2.1 is a useful non-invasive diagnostic tool for H. pylori infection in children over 6 years of age. PMID- 12745465 TI - Predicting urine culture results by dipstick testing and phase contrast microscopy. AB - AIMS: Urine is the most frequently received clinical specimen for bacterial culture. To determine whether dipstick or microscopy results reliably predicted the presence or absence of a reportable urinary pathogen we performed dipstick testing and phase contrast microscopy on unspun urine from 500 specimens. We also investigated the relationship between the presence of squamous epithelial cells (SECs) and mixed growth on culture. METHODS: For each specimen, the presence of leukocyte esterase and urinary nitrite was recorded as well as the quantity of leukocytes (WBC), SECs, and erythrocytes. All cultures had the number and type of each organism recorded. Pyuria was defined as > or =10 WBC/mm(3). RESULTS: Specimens with <10 SEC/mm(3) had fewer isolates (0.9 isolate per culture) than specimens with > or =10 SEC/mm(3) (two isolates per culture); therefore, SEC contamination was defined as > or =10 SEC/mm(3). Of 500 specimens, 266 (53%) had no growth and 136 (27%) had mixed growth. A total of 288 (58%) specimens had negative dipstick results and completely normal microscopy. Of these, 11 (4%) had a pure growth of a urinary pathogen but the pathogen was present in only three (1%) at > or =10(5) CFU/mL. Of 413 urine specimens without SEC contamination, 323 (78%) had either no detectable growth or pure growth compared with only 41 of 87 (47%) with SEC contamination (P<0.001). Of the 413 urine specimens without SEC contamination, 90 (22%) had > or =2 organisms compared with 46 of 87 (53%) with contamination (P<0.01). Pyuria was present in only 21 of 266 (8%) urine specimens without growth but was present in 60 of 95 (64%) specimens containing a reportable pathogen (P<0.01). CONCLUSIONS: SEC contamination does not reliably predict cultures with mixed growth, urine specimens with negative dipstick results and microscopy rarely contain a reportable urinary pathogen, and screening algorithms are warranted and justified. PMID- 12745468 TI - Testicular epidermoid cyst: a case of triple pathology. PMID- 12745466 TI - Test and teach. Number fifty-three. Diagnosis: Calcifying fibrous pseudotumour. PMID- 12745469 TI - Delayed renal allograft failure due to polyomavirus-associated tubulointerstitial nephritis. PMID- 12745470 TI - Triplet pregnancy with a co-existing hydatidiform mole, two liveborn infants and a single placenta. PMID- 12745471 TI - Cotyledonoid dissecting leiomyoma. PMID- 12745472 TI - Gastrointestinal small vessel vasculitis in a patient with crescentic glomerulonephritis and longstanding idiopathic neutropaenia treated with G-CSF. PMID- 12745473 TI - Publications on pathology by Australian laboratories. PMID- 12745474 TI - Re: Lymph node sampling for flow cytometric analysis. PMID- 12745483 TI - What's new in Nicotine & Tobacco Research? PMID- 12745484 TI - Introduction: studies of tobacco dependence treatment and methodology. PMID- 12745485 TI - My career in psychopharmacology. PMID- 12745486 TI - A minimum 6-month prolonged abstinence should be required for evaluating smoking cessation trials. PMID- 12745487 TI - Effect of smoking reduction on later cessation: a pilot experimental study. AB - It is unclear whether reducing the number of cigarettes in smokers not trying to quit increases or decreases the likelihood of future quitting. In a pilot study, smokers not currently interested in quitting (n=67) were randomized to two groups. Experimental participants received behavioral treatment and nicotine replacement therapy (choice of gum, patch, or inhaler) to reduce smoking by 50% over 4 weeks, followed by brief advice to quit. Usual-care participants received only brief advice to quit and nicotine replacement if they decided to quit. During the 4-week treatment period, nonabstaining reduction participants decreased from 23 to 14 cigarettes per day (p<.01) and maintained their reduction over the 6-month follow-up period. At the 6-month follow-up, 35% of usual-care and 41% of reduction participants (nonsignificant [ns]) moved forward in their stage of change. Over the 6 months, 34% of usual-care participants had at least one 24-h quit attempt, compared with 25% of reduction participants (ns). A total of 9% of usual-care participants remained quit at 6 months vs. 13% in the reduction group (ns). These preliminary results suggest that adding a reduction option neither increases nor undermines interest in cessation. Higher than expected rates of attempted cessation and quitting in the usual-care group suggest that we recruited smokers whose motivation to quit was above average. Thus, a replication test in a less-motivated group of smokers is needed. PMID- 12745488 TI - Discussion of NRT and other antismoking interventions in UK general practitioners' routine consultations. AB - Nicotine replacement therapy (NRT) is an effective smoking cessation treatment, but little information is available about how primary care physicians use the therapy. We investigated the quality of UK general practitioners' (GPs') advice against smoking, including how frequently they recommend NRT to smokers. We compared the characteristics of smokers whom GPs recommended use NRT with smokers who did not receive GP recommendations. We conducted a cross-sectional survey of 3,525 adult patients attending 35 UK GPs over a 16-month period. Of those surveyed, 2,955 eligible patients (83.8%) completed preconsultation questionnaires, and 1,026 (34.7%) were regular smokers, with 927 (90.4%) of the regular smokers completing postconsultation questionnaires. Of those completing the questionnaires, 20.2% (95% CI, 17.7% to 22.9%) or 187 recalled discussion of smoking with their GPs and of these, 66.3% (95% CI, 60.1% to 74.1%) or 124 perceived that they had been given an unequivocal message to stop smoking. Although 60.4% of smokers (606 of 1,004) (95% CI, 57.3% to 63.3%) or smoked more than 10 cigarettes daily, only 17.6% (95% CI, 12.9% to 23.7%) or 33 of those recalling antismoking advice reported discussion of NRT. Accordingly, GPs discussed NRT with only a minority of smokers who might benefit from the therapy. Intending to give up smoking in the next 4 weeks was the only variable independently associated with smokers' recall of discussing NRT (OR=2.58 [95% CI 1.20% to 5.57%]). The study findings thus provide only limited information to support the notion that UK GPs recommend NRT in an evidence-based manner. Now that NRT is available by prescription from UK GPs, further research is needed to monitor whether this effective antismoking therapy is being prescribed appropriately. PMID- 12745489 TI - Effects of the nicotine patch on performance during the first week of smoking cessation. AB - Complaints of feeling unfocused and being unable to concentrate are common during smoking cessation, and such feelings may contribute to a subtle erosion of the motivation to quit. A heterogeneous sample of 21 established smokers (10 women, 11 men) completed this double-blind, placebo-controlled study to test the hypothesis that nicotine replacement during cessation therapy (using a 21-mg nicotine patch) would improve performance on tasks sensitive to nicotine deprivation. Participants were trained to stable performance on simple reaction time, mathematical processing, Sternberg memory, rapid visual-information processing, grammatical reasoning, and the Stroop Color-Word (Stroop) tasks. They received smoking cessation counseling and were randomly assigned to nicotine patch and placebo patch groups. Performance was assessed prior to cessation, and early (days 2 and 3) and late (days 5 through 7) in the first cessation week. The hypothesis was not supported. Increased accuracy was associated with the patch only for grammatical reasoning. No reaction time differences were found in the simple reaction time, grammatical reasoning, and mathematical processing tasks. Reaction time was faster in the placebo group on the more difficult portions of those tasks requiring sustained attention (rapid visual-information processing, Stroop Color-Word, and Sternberg memory tasks). These results differ substantially from those obtained when young adults are allowed to smoke or chew nicotine gum after relatively brief periods of deprivation. Evidence that smoking may interfere with cognition is accumulating; these results support this view. The subjective performance decrements noted by many smokers during cessation may be related to overall negative affect, rather than to direct effects on cognition and attention. PMID- 12745490 TI - Comparison of nicotine patch alone versus nicotine nasal spray alone versus a combination for treating smokers: a minimal intervention, randomized multicenter trial in a nonspecialized setting. AB - This multicenter, randomized, open-label clinical trial was conducted to determine whether the combined use of nicotine patch therapy and a nicotine nasal spray would improve smoking abstinence rates compared to either treatment alone, without behavioral counseling. Data were collected at 15 regional cancer control oncology centers within the North Central Cancer Treatment Group. Of the 1384 smokers randomized to the study, 20% were abstinent from smoking at 6 weeks and 8% were abstinent at 6 months. At 6 weeks, the 7-day point prevalence smoking abstinence rate for the patch alone (21.1%) was superior to the spray (13.6%) but was significantly lower than the rate for combination therapy (27.1%). At 6 months, the 7-day point prevalence abstinence rates were not significantly different among the three groups. Combination nicotine nasal spray and nicotine patches were delivered safely in a nonspecialized outpatient clinical setting and enhanced short-term smoking abstinence rates, but these rates were not sustained at 6 months. PMID- 12745491 TI - Evaluation of an Internet-based smoking cessation program: lessons learned from a pilot study. AB - The potential contribution of the Internet to smoking cessation seems huge, given that a majority of Americans now have both computers and telephones. Despite the proliferation of Web sites offering smoking cessation support, there is little empirical evidence regarding the efficacy of Internet-delivered cessation programs. We developed a cessation Web site and conducted a short-term evaluation of it, examining recruitment approaches, Web site use patterns, alternative retention incentives and re-contact modes, satisfaction, and cessation rate. The intervention included modules on social support and cognitive-behavioral coping skills configured to take advantage of the interactive and multimedia capabilities of the Internet. Cessation and satisfaction data were obtained from a subsample of 370 subjects followed for 3 months. The program was rated as easy to use, and the social support group component was used most frequently. The cessation rate (abstinence for the previous 7 days) at 3 months was 18%, with nonrespondents (n=161) considered smokers. Among a variety of traditional and Internet-based recruitment strategies, the most successful made use of Internet user groups and search engines. Methodological and procedural issues posed in conducting research on the Internet are discussed. PMID- 12745492 TI - Comparison of the effects of combined nicotine replacement therapy vs. cigarette smoking in males. AB - This open study assessed the effects of nicotine replacement therapy (NRT) on the normalizing of exhaled carbon monoxide (CO), transcutaneous partial oxygen tension (tcpO(2)), plasma cotinine and thiocyanate levels, and cardiovascular risk markers in abstinent subjects compared with untreated smokers after 4, 8, 12, and 26 weeks. The trial enrolled 197 subjects in two parallel groups: 164 subjects who received NRT (patch plus gum) for 12 weeks and 33 untreated smokers (controls). At 26 weeks, 123/164 participants in the treatment group had completed the study; 51/123 (41.5%) sustained abstinence from smoking, whereas 72/123 (58.5%) had relapsed. Changes in cotinine (abstainers: 291.6 ng/ml at baseline vs. 27.3 ng/ml at week 26; p<.0001) and thiocyanate levels (abstainers: 10.4 ng/ml at baseline vs. 6.2 ng/ml at week 26; p<.0001) and expired CO (abstainers: 30.4 ppm at baseline vs. 4.2 ppm at week 26; p<.0001) accurately reflected the changes in smoking and/or NRT use in both abstainers and relapsers. After they stopping smoking, tcpO(2) significantly improved in abstainers (34.9 mmHg at baseline vs. 50.4 mmHg at week 26; p<0.0001). Inverse correlations between the number of daily cigarettes and plasma cotinine, thiocyanate, and exhaled CO levels were observed in both relapsers and smokers. A clinically significant increase in HDL cholesterol (39.0 vs. 44.7 mg/dl; p<.0001) occurred in the abstainers between baseline and study end. Use of combination NRT to achieve abstinence resulted in marked improvements in biochemical parameters in abstainers and partial improvements in relapsers. The safety of combination NRT was confirmed by the absence of overdose-related adverse events. PMID- 12745493 TI - A contingent payment model of smoking cessation: effects on abstinence and withdrawal. AB - The present study was designed to characterize nicotine withdrawal during a 5-day period in which smokers who were not trying to quit were offered monetary incentives to abstain while residing in their usual environments. Participants were randomly assigned to one of three groups. In two groups, monetary payment was delivered contingent on breath carbon monoxide levels (CO< or =8 ppm) indicating recent smoking abstinence, with the amount of payment differing between the two groups. The third group was a control group in which payment was delivered independent of smoking status. Participants provided CO samples three times per day (morning, afternoon- and evening) for 5 days (Monday-Friday). At each evening visit, all participants completed a nicotine withdrawal questionnaire and other questionnaires. Contingent payment significantly decreased expired-air CO and salivary cotinine levels as compared with the control group. No significant differences in abstinence were noted as a function of the amount paid. Participants in both contingent payment groups reported significantly more withdrawal symptoms than those in the noncontingent control group, including increases in anxiety and nervousness, impatience and restlessness, hunger, and desire to smoke. Such contingent payment procedures may provide an effective method for studying nicotine withdrawal in smokers that does not require the costly and inconvenient practice of housing research participants on a closed ward to prevent smoking. PMID- 12745494 TI - Predictors of quitting in hospitalized smokers. AB - Hospitalization represents a teachable moment for quitting. The current study examined predictors of quitting among hospitalized smokers. Patients reported smoking history and demographic characteristics during in-hospital baseline interviews. Discharge diagnosis also was collected. Smoking status was ascertained in interviews at 7 days and at 12 months after discharge. A total of 2,350 patients in four Minneapolis and St. Paul (Twin Cities), Minnesota, area hospitals participated in the study; 1,477 patients who provided data at both follow-ups and whose 12-month self-report of quitting was corroborated by cotinine analysis of saliva samples were included in the current analyses. Predictors of both short- and long-term abstinence in the multivariate analysis included smoking-related illness, age (those who were older were more likely to be abstinent), stage of change (precontemplators were least likely to quit, and those initially in action were most likely to quit), and time to first cigarette (those who reported smoking within 5 min of awakening were least likely to quit). The predictors presented few surprises; the most important finding may have been that the experience of hospitalization itself led to substantial long-term quitting for virtually all categories of hospitalized smokers. PMID- 12745495 TI - Efficacy of oral transmucosal nicotine lozenge for suppression of withdrawal symptoms in smoking abstinence. AB - Nicotine replacement therapy relieves withdrawal symptoms, significantly improving smoking cessation rates. Oral transmucosal nicotine (OT-NIC) is a novel nicotine delivery system consisting of a lozenge (OT-NIC unit) containing 4 mg of nicotine, which is dissolved in the cheek pouch, releasing nicotine for absorption through the buccal mucosa. Theoretical advantages of OT-NIC include that it does not require special chewing methods or interfere with dental work, it provides sensory oral effect, and it can be dosed to effect. This study aimed to determine the preliminary safety and efficacy of OT-NIC for suppression of nicotine withdrawal symptoms over 8 days of smoking abstinence and to assess flavor preference, sensory characteristics, and acceptability. In an open-label, within-subjects design, 11 smokers used three different flavors of OT-NIC ad lib in response to withdrawal symptoms and craving. On days 1 and 8, withdrawal symptoms were measured with pre- and postadministration scores for each of the first three OT-NIC units used (three different flavors presented in random order). Flavor preference, desire to smoke, sensory characteristics, acceptability, and overall OT-NIC performance were rated at the end of each day. A generalized estimating equation analysis was conducted to account for the repeated-measures design. Use of OT-NIC resulted in a significant decrease in a composite withdrawal score aggregating scores from all eight symptoms (decline of.57 units in a possible range of 4, p<.01). Withdrawal scores for anxiety, craving, difficulty concentrating, impatience, and restlessness significantly decreased. Depressed mood, increased appetite, and irritability did not change. Order of flavor preference was peppermint over cinnamon over plain. Acceptability and ratings for sensory characteristics were favorable, and OT-NIC was well tolerated with no serious adverse effects. In conclusion, OT-NIC showed promise as a potential new aid for smoking cessation. PMID- 12745496 TI - Naltrexone treatment for alcoholics: effect on cigarette smoking rates. AB - Naltrexone (NTX), by its pharmacological action in the mesolimbic pathways, should decrease reinforcement from nicotine as well as from alcohol. By means of this mechanism, NTX could result in temporary increases in smoking followed by decreased smoking rates among alcoholics not motivated to quit smoking. The change from pretreatment in smoking rates of 73 recently abstinent alcoholics in a 12-week clinical trial of NTX vs. placebo during alcoholism treatment was compared during 8 of the 12 weeks. Only smokers compliant with NTX were included in the analyses. NTX was associated with decreased smoking at every time point, but the effect was significant at only one time point. When alcohol relapsers were excluded, NTX patients showed decreased smoking at every time point, but the effect was significant at only two time points, a reduction of about five cigarettes per day. When smoking stage of change was included in the analyses, NTX showed no significant main or interaction effects on smoking rate. Precontemplators showed significantly less change in smoking rate than all other patients at the first and last four time points. Therefore, NTX alone currently does not show promise for promoting smoking reduction among recently abstinent alcoholics who have not sought or been given smoking cessation treatment. Further research is needed on possible effects with smokers motivated to quit smoking and on other methods of promoting smoking cessation among alcoholics. PMID- 12745497 TI - The demand for nicotine replacement therapies. AB - This paper reports on the first econometric study to examine the determinants of nicotine replacement therapy (NRT) demand. Pooled cross-sectional time-series scanner-based data for 50 major metropolitan markets in the United States covering the period between the second quarter 1996 and the third quarter 1999 are used in the analysis. Fixed-effects modeling is employed to assess the impact of NRT prices, cigarette prices, and other determinants on NRT demand. The estimates indicate that decreases in the price of NRT and increases in the price of cigarettes would lead to substantial increases in per-capita sales of NRT products. The average own-price elasticity of demand, which measures the responsiveness of consumer demand for NRT products to changes in the price of NRT, for Nicoderm CQ and Nicorette, is -2.33 and -2.46, respectively. The average cross-price elasticity of demand, which measures the responsiveness of consumer demand for NRT products to changes in the price of cigarettes, for Nicoderm CQ and Nicorette, is 0.772 and 0.764, respectively. PMID- 12745498 TI - The heritability of failed smoking cessation and nicotine withdrawal in twins who smoked and attempted to quit. AB - The magnitude of the association between nicotine withdrawal and failed smoking cessation remains uncertain, and the potential genetic contribution to that relationship is unknown. We used a twin design study to test if genetic and environmental risk factors contribute to nicotine withdrawal and failed cessation. We then estimated the magnitude of genetic and environmental risk factors and the associated correlations. Cigarette consumption data were obtained in 1992 from 4,112 (1,806 pairs, 496 singletons) male twin pairs from the Vietnam Era Twin Registry. Participants responded to a telephone administration of the Diagnostic Interview Schedule. Odds ratios were computed for failed smoking cessation as a function of 12 individual nicotine withdrawal symptoms. Genetic model fitting was performed on a sample of 1818 twin pairs, all of whom were lifetime smokers; both twins of the pairs had made at least one quit attempt. After adjustment for cigarette consumption, odds of failed smoking cessation ranged from.87 (95% CI.76-.99) for "increased appetite/weight gain" to 1.85 (95% CI 1.55-2.22) for "depressed." Genetic influences accounted for 54% and 29.7% of the variance in risk for failed smoking cessation and nicotine withdrawal, respectively. The correlation between genetic influences was significant (r=.31, 95% CI.17-.45). The magnitude of the association between failed cessation and nicotine withdrawal varied by symptom. The risk of experiencing nicotine withdrawal after a quit attempt was related partly to genetic vulnerability; lifetime risk for both failed cessation and nicotine withdrawal was related partly to genetic factors that were correlated. PMID- 12745500 TI - What's new in Nicotine & Tobacco Research? PMID- 12745499 TI - A factor analysis of the Fagerstrom Test for Nicotine Dependence (FTND). AB - Psychometric study of the Fagerstrom Test for Nicotine Dependence (FTND) provides insight into its structure and the dimensions of nicotine addiction it assesses. We evaluated the factor structure of the FTND in 541 research volunteers, most with histories of polysubstance use. Tetrachoric and phi correlation techniques were utilized and promax- and varimax-rotated solutions are reported. Two factors were found. Factor 1 was defined by questions regarding time to first cigarette in after waking, which cigarette is most preferred, and prominence of morning smoking. Factor 2 was defined by questions regarding difficulty refraining from smoking, amount smoked, and smoking while ill. The question "How soon on waking do you smoke your first cigarette?" loaded substantially on both Factor 2 and Factor 1. Repeating the analyses after stratification by gender did not change the results. The factor structure from our sample population was similar to results reported by previous studies with different types of populations. We propose that Factor 1 assesses the degree of urgency to restore nicotine levels to a given threshold after nighttime abstinence, whereas Factor 2 reflects the persistence with which nicotine levels are maintained at about that threshold during waking hours. Thus, the FTND may assess distinguishable self-reportable pharmacological dimensions of nicotine addiction. These two dimensions may provide indirect assessment of a smoker's daily nicotine intake. Testing this hypothesis requires correlation of biomarkers and responses to specific items of the FTND. PMID- 12745501 TI - The need for dissemination of evidence-based results from research on nicotine and tobacco. PMID- 12745502 TI - Does menthol enhance the addictiveness of cigarettes? An agenda for research. PMID- 12745503 TI - Measures of abstinence in clinical trials: issues and recommendations. AB - A workgroup formed by the Society for Research on Nicotine and Tobacco reviewed the literature on abstinence measures used in trials of smoking cessation interventions. We recommend that trials report multiple measures of abstinence. However, at a minimum we recommend that trial: (a) report prolonged abstinence (i.e., sustained abstinence after an initial period in which smoking is not counted as a failure) as the preferred measure, plus point prevalence as a secondary measure; (b) use 7 consecutive days of smoking or smoking on > or = 1 day of 2 consecutive weeks to define treatment failure; (c) include non-cigarette tobacco use, but not nicotine medications in definitions of failure; and (d) report results from survival analysis to describe outcomes more fully. Trials of smokers willing to set a quit date should tie all follow-ups to the quit date and report 6- and/or 12-month abstinence rates. For these trials, we recommend an initial 2-week grace period for prolonged abstinence definitions; however, the period may vary, depending on the presumed mechanism of the treatment. Trials of smokers who may not be currently trying to quit should tie follow-up to the initiation of the intervention and should report a prolonged abstinence measure of > or = 6-month duration and point prevalence rates at 6- and 12-month follow ups. The grace period for these trials will depend on the time necessary for treatment dissemination, which will vary depending on the treatment, setting, and population. Trials that use short-term follow-ups (< or = 3 months) to demonstrate possible efficacy should report a prolonged abstinence measure of > or = 4 weeks. We again recommend a 2-week grace period; however, that period can vary. PMID- 12745504 TI - The increasing recalcitrance of smokers in clinical trials II: Pharmacotherapy trials. AB - Irvin and Brandon (Nicotine & Tobacco Research, 2, 79-84, 2000) reported a significant decline in reported abstinence rates between 1977 and 1996 from clinical trials of cognitive-behavioral smoking cessation treatments based on coping skills training. The present study extends that approach to the analysis of pharmacotherapy trials. A literature search identified 59 studies, published between 1983 and 2000 and conducted in the U.S., that reported post-cessation abstinence rates after treatment with nicotine gum, nicotine patch or any type of placebo medication. Across all three types of treatment conditions and four post cessation assessment points, negative correlations between publication year and abstinence rates were found. The strongest pattern of negative correlations was found for the placebo conditions. However, the correlations for placebo conditions could be accounted for by the simultaneous shift toward treatments offered in individual rather than group format. No other methodological or subject variable appeared to mediate the declining outcomes. Findings are discussed with respect to the theory that the population of remaining smokers is becoming progressively more dependent and difficult to treat. PMID- 12745505 TI - Hardening and the hard-core smoker: concepts, evidence, and implications. AB - A nascent debate pits researchers who believe that hard-core smokers are coming to dominate the remaining population of smokers against others who perceive the hardening of the target as a far more distant concern. At stake is the future emphasis of tobacco control: should we alter the current allocation of resources between treatment of individual smokers and modification of the psychosocial environment through public education and policy measures? We review the evidence and conclude that: (1) hardening is probably occurring in the sense that, compared with earlier generations, many of today's smokers possibly do have greater difficulty quitting, or are inherently less willing to do so. (2) Hardening may be most usefully construed in the context of specific groups of smokers, such as the mentally ill, who may constitute a growing fraction of the remaining smoking population. (3) Using conventional measures, however, we find little evidence that the population of smokers as a whole is hardening. Cessation rates have not decreased. (4) Truly hard-core smokers necessarily constitute a very small fraction of the population. Quitting-susceptible smokers continue to dominate the smoking population. (5) Hardening and the potential existence of true hard-core smokers recommend creative thinking about, and devotion of resources to, finding new ways to help the most dependent smokers to quit. (6) Sound research recommends the expansion of comprehensive tobacco-control programs in both the public and private sectors, and does not support reallocation of resources from such programs toward more intensive individualized treatment. We can afford both. PMID- 12745506 TI - Nicotine withdrawal and depressive symptomatology during short-term smoking abstinence: a comparison of postmenopausal women using and not using hormone replacement therapy. AB - This study investigated whether taking medications for transdermal hormone replacement therapy (HRT) influenced smoking-cessation variables in postmenopausal women undergoing short-term abstinence from cigarettes. Women were recruited into two groups according to their pre-enrollment medication status- those currently on HRT (n = 17) or those not on HRT (n = 13). The HRT group had their previous medication replaced with a standard 0.1 mg estradiol transdermal system and 2.5 mg of Cycrin daily. After 2 weeks of medication adjustment, participants continued smoking as usual for 1 week, at which time baseline measurements were taken. Participants were then instructed to quit smoking for the remaining 2 weeks. They were provided with smoking-cessation counseling and monitored for abstinence. Data were collected during five clinic visits on all dependent measures: Minnesota Nicotine Withdrawal Scale, Beck Depression Inventory (BDI) scale, Profile of Mood States, Motor Speed Tasks, and Reaction Time Test. Contrary to our hypothesis, the exogenous hormone use did not have a differential effect on most of the dependent variables during the first 2 weeks of smoking abstinence. One exception was depressive symptomatology: the BDI change scores (week 2 - baseline) differed significantly for the HRT and non-HRT groups (p = .045), with women in the HRT group experiencing an increase in depressive symptomatology. This finding, though preliminary, may have clinical implications for postmenopausal women who attempt to quit smoking while on HRT, particularly since depressed mood following abstinence is associated with a relapse to smoking. PMID- 12745507 TI - Depressive symptoms and smoking cessation among inner-city African Americans using the nicotine patch. AB - This study examined the relationship between depressive symptoms and smoking cessation among a sample of inner-city African American smokers using the nicotine patch. Analyses were conducted on data from a previous randomized trial that tested the effects of culturally sensitive vs. standard self-help quitting materials. The study sample consisted of 498 African American smokers (mean age = 42.95, SD = 10.40; 60% female) recruited from a large hospital. Participants in both groups received 8 weeks of nicotine replacement therapy. Level of smoking, quit status, and depressive symptoms (Medical Outcomes Survey Short Depression Screen) were assessed at baseline, week 4 (mid-treatment), and 6-month follow-up. Analyses that controlled for randomization group generally did not support the hypothesis that baseline levels of depressive symptoms predict smoking at Week 4 or Month 6. Cross-sectional analyses at each time point indicated that depressive symptoms were positively associated with smoking level (both: beta = 0.24, p < .05). Changes (increases) in depressive symptoms from baseline to week 4 predicted higher smoking levels at follow-up (beta = 0.19, p < .05). Although the results indicated a significant association between depressive symptoms and level of smoking, they generally did not support the notion that depressive symptoms are associated with failure to quit. PMID- 12745508 TI - A history of depression and smoking cessation outcomes among women concerned about post-cessation weight gain. AB - Because a history of depression has been hypothesized to affect cessation efforts and may be particularly problematic for women concerned about weight gain, we sought to document the prevalence of depression history among weight-concerned women smokers and evaluate its effect on treatment outcome. We also evaluated the impact of baseline depressive symptoms and cessation-related changes in symptoms. Women (N = 219) were classified as depression history positive (Major Depressive Disorder [MDD]) (MDD+) or negative (MDD-) according to responses on the Inventory to Diagnose Depression-Lifetime Version. All women received a group-based smoking cessation treatment. Women provided expired-air carbon monoxide samples, completed measures of depressive symptoms, and were weighed at pretreatment and 1, 3, 6, and 12 months after quitting. Fifty-two per cent (n = 115) reported a lifetime history of major depressive disorder. Although MDD+ women were significantly more nicotine dependent, rates of continuous abstinence did not differ between MDD+ and MDD- women. However, MDD+ women were more likely to drop out of treatment prior to quitting. Additionally, depressive symptoms were associated with abstinence irrespective of depression history. Women who reported an increase in depressive symptoms from pre- to post-treatment were significantly less likely to be abstinent post-treatment, suggesting that depressive symptoms are more predictive of outcome than is previous disorder. Moreover, because of the prevalence of depression history among this subgroup of women smokers and its impact on early attrition, additional engagement and retention strategies may be useful. PMID- 12745509 TI - A study of depressive symptoms and smoking behavior in adult male twins from the NHLBI twin study. AB - Self-report measures of depressive symptoms, such as the Center for Epidemiological Studies-Depression Scale (CES-D), correlate with current and lifetime smoking status. In one previous study of adult female twins, genetic factors accounted for the covariation of liability to a diagnosis of major depressive disorder and liability to lifetime smoking (Kendler, Neale, MacLean, Heath, Eaves, & Kessler, 1993b, Archives of General Psychiatry, 50, 36-43); however, it remained unclear whether genetic effects also account for the covariation between subclinical depressive symptomology and smoking behavior. In this study, we use twin structural equation modeling to explore whether genetic and/or environmental influences contribute to the covariation between depressive symptoms, as measured by the CES-D, and current and lifetime smoking status among 120 monozygotic and 114 dizygotic Caucasian male twin pairs (aged 59-69). In this sample, depressive symptoms showed small but significant correlations with current and lifetime smoking status. Univariate twin analyses indicated that additive genetic and non-shared environmental factors contributed significantly to liability to current and lifetime smoking. However, the majority of variance in CES-D scores was attributable to non-shared (individual) environment. In bivariate analyses, non-shared environmental factors accounted for the majority of covariation between liability to depressive symptoms (CES-D scores > or = 8; above the 75th percentile) and liability to current and lifetime smoking status. Taken together with the previous literature, these results suggest that the etiology of covariation among depressive symptoms and smoking behavior may vary by measurement and severity of depressive symptomology. PMID- 12745510 TI - Psychiatric disorders, familial factors and cigarette smoking: I. Associations with smoking initiation. AB - The aims of this study were to examine associations between smoking initiation (five cigarettes lifetime) and lifetime psychopathology, regular smoking by family members, and psychopathology in family members; to describe the degree to which the onset of the disorder precedes or follows smoking initiation; and to examine whether smoking initiators differ as a function of age of smoking onset. Nine hundred and forty-one participants were interviewed at three time points, beginning in high school and most recently at age 24. Lifetime psychiatric diagnoses were obtained at each assessment, as were data regarding smoking initiation. Biological parents and full siblings were interviewed for lifetime psychopathology and regular smoking. Most measures of lifetime psychopathology were associated with smoking initiation. Rates of initiation were especially elevated in participants with multiple disorders. Regular smoking by mother and a sibling (but not father) was associated significantly with smoking initiation, as were two of four measures of psychopathology in relatives. When all significant univariate variables were examined in a single model, drug use disorders, regular smoking by mother, and regular smoking by a sibling remained significantly associated with smoking initiation. Smoking initiation preceded approximately half of the examined diagnostic categories. Eleven variables differentiated early vs. late smoking initiators. Several interactions with gender were found. In every instance, smoking initiation was more strongly associated with the risk factor for young women than for young men. To our knowledge, this is the first time that the relations of psychopathology and familial factors with smoking initiation have been examined simultaneously or in this much detail. Results underscore the potential importance of assessing and treating psychiatric disorders in smoking prevention and cessation efforts. PMID- 12745512 TI - Gender differences in quit rates following smoking cessation with combination nicotine therapy: influence of baseline smoking behavior. AB - Women are less successful than men at quitting smoking. We examined whether the male vs. female cessation outcome was influenced by baseline smoking behavior in participants who attempted to quit by using nicotine inhaler (NI) plus nicotine patch (NP) combination therapy. This double-blind, randomized, placebo-controlled trial enrolled 196 men and 204 women. Group 1 (99 men, 101 women) received NI plus NP (15 mg nicotine/16 h) for 6 weeks, then NI plus placebo patch (PP) for 6 weeks, then NI alone for 14 weeks. Group 2 (97 men, 103 women) received NI plus PP for 12 weeks, then NI alone for 14 weeks. Outcome measures were continuous self-reported abstinence and expired carbon monoxide concentration < 10 ppm. Baseline nicotine dependence was assessed by the Fagerstrom Test for Nicotine Dependence (FTND), and behavioral dependence by the 18-question Glover-Nilsson Smoking Behavioral Questionnaire (GN-SBQ). Male vs. female complete abstinence rates, regardless of treatment group, were 61.7% vs. 46.6% at 6 weeks (p = .0022), 42.3% vs. 30.9% at 12 weeks (p = .017), 30.1% vs. 17.6% at 6 months (p = .003), and 23.0% vs. 10.8% at 12 months, respectively (p = .001). Men had significantly higher baseline FTND (p = .0180) and lower total GN-SBQ (p < .0001) scores than women. In conclusion, women appear to have higher behavioral, and lower nicotine, dependence than men according to the GN-SBQ and the FTND; thus both nicotine and behavioral treatment should be tailored to women to increase their chances of abstinence. PMID- 12745511 TI - Targeting smokers at increased risk for relapse: treating women and those with a history of depression. AB - Some studies have shown that female smokers and smokers with a history of depression have an increased risk of relapse following smoking cessation treatment. This study examined the efficacy of bupropion sustained-release (SR) and the nicotine patch for smoking cessation in subgroups of smokers at possible risk for relapse. Data for this study were from a previously published randomized, double-blind, placebo-controlled clinical trial in which 893 smokers were randomized to four treatment conditions: placebo tablet + placebo patch, placebo tablet + 21 mg/24-hr nicotine patch, 300mg bupropion SR + placebo patch, and 300mg bupropion SR + 21 mg/24-hr nicotine patch. Study medication continued for 8 weeks after the quit day; brief individual cessation counseling was provided during weekly clinic visits. In comparison to the placebo tablet, bupropion SR approximately tripled 1-year non-smoking rates among women and previously depressed individuals. In contrast, the nicotine patch did not significantly improve cessation rates for any group. We conclude that bupropion SR is a first-line treatment for smoking that has the potential to benefit all smokers, especially women and the previously depressed. PMID- 12745513 TI - Laternal nicotine exposure increases nicotine preference in periadolescent male but not female C57B1/6J mice. AB - Maternal cigarette smoking is a risk factor for adolescent smoking. One possible explanation for increased smoking by human adolescents after maternal nicotine exposure is that exposure increases nicotine preference. However, it is difficult to separate the biological and social causes of smoking behavior in humans. This experiment examined the relationship between maternal nicotine exposure and nicotine preference in periadolescent offspring using a mouse model of oral nicotine consumption. Pregnant females were provided saccharin-flavored water containing 50 microg/ml nicotine (n = 4) or no nicotine (n = 5) from the ninth day of gestation through weaning on postnatal day (PD) 21. Offspring from these females were tested for nicotine preference during periadolescence (PDs 35-42) by providing access to both saccharin-only and nicotine solutions (50 microg/ml) 24 hr a day in the home cage in a two-bottle choice test. Male mice exposed maternally to nicotine (n = 9) exhibited an increased nicotine preference in adolescence compared to non-nicotine exposed controls (n = 12). Maternal nicotine exposure did not alter nicotine preference by periadolescent female mice. Nicotine consumption was confirmed by serum cotinine measurement. These data are consistent with human epidemiological reports that maternal nicotine exposure is associated with increased risk of cigarette smoking. Differential outcomes for males and females suggest that different processes underlie sex differences in nicotine consumption following maternal nicotine exposure. PMID- 12745514 TI - The effect of smoking on endothelial vasodilatory function evaluated by local infusion of metacholine in the forearm is dependent on the duration of smoking. AB - The objective of the study was to evaluate if endothelial vasodilatory function in the human forearm is impaired by regular cigarette smoking. The setting was a tertiary university hospital. Subjects were 56 apparently healthy subjects from a population screening (mean age 50) and 52 young healthy volunteers (mean age 25) who were investigated regarding endothelial-dependent (EDV) and endothelial independent vasodilation (EIDV) by means of local infusion of metacholine (MCh; 2 and 4 mg/min) and sodium nitroprusside (SNP; 5 and 10 mg/min) in the forearm. Forearm blood flow (FBF) was measured by venous occlusion plethysmography. The MCh to SNP FBF ratio was denoted the endothelial function index. In the young subjects, no differences between smokers (n = 12) and non-smokers regarding EDV or EIDV were seen. In the population sample, however, the smokers (n = 8) showed an attenuated endothelial function index when compared with non-smokers (1.0 +/- 0.1 vs. 1.3 +/- 0.3, p = .02). The EDV showed a significant inverse relationship to the duration of smoking (r = -0.52, p < .05), independent of age, when the smokers in both groups were analyzed together. A similar, although not significant, relation was found between the endothelial function index and the duration of smoking (p = -.44). The present study showed that endothelial vasodilatory function was impaired in middle-aged, but not young, smokers, suggesting that the duration of smoking is of major importance for the deleterious effects of smoking on endothelial vasodilatory function. PMID- 12745516 TI - Twelve tips to improve bedside teaching. AB - Bedside teaching has long been considered the most effective method to teach clinical skills and communication skills. Despite this belief, the frequency of bedside rounds is decreasing and it is believed that this is a major factor causing a sharp decline in trainees' clinical skills. Several barriers appear to contribute to this lack of teaching at the bedside and have been discussed extensively in the literature. Concern about trainees' clinical skills has led organizations such as the American Council for Graduate Medical Education (ACGME) and the WHO Advisory Committee on Medical training to recommend that training programs should increase the frequency of bedside teaching in their clinical curricula. Although obstacles to bedside teaching are acknowledged, this article in the '12 tips' series is a detailed description of teaching strategies that could facilitate a return to the bedside for clinical teaching. PMID- 12745515 TI - Seizure associated with sleep deprivation and sustained-release bupropion. AB - This case report describes a generalized seizure associated with sustained release bupropion use and sleep deprivation. The subject, a 31-year-old female smoker, was participating in a clinical trial evaluating an investigational medication for smoking cessation that used sustained-release bupropion as an active control. After 5 weeks of bupropion use, the subject experienced a generalized tonic clonic seizure after staying up nearly all night packing and moving to a new residence. The patient had no other risk factors for seizures. We suggest that sleep deprivation may add to the risk of bupropion-associated seizures. PMID- 12745517 TI - A qualitative study of constructive clinical learning experiences. AB - Little is known about the effectiveness of clinical education. A more educational structure is considered to be potentially beneficial. The following structured components were added to a surgical clerkship: logbooks, an observed student patient encounter, individual appraisals, feedback on patient notes, and (case) presentations by students. The authors organized two focus-group sessions in which 19 students participated to explore their perceptions about effective clinical learning experiences and the newly introduced structured components. The analysis of the transcripts showed that observation and constructive feedback are key features of clinical training. The structured activities were appreciated and the results show the direction to be taken for further improvement. Learning experiences depended vastly on individual clinicians' educational qualities. Students experienced being on call, assisting in theatre and time for self-study as instructive elements. Recommended clerkship components are: active involvement of students, direct observation, selection of teachers, a positive learning environment and time for self-study. PMID- 12745518 TI - Teaching at the bedside: a new model. AB - The patient is at the center of clinical medicine. In order to effectively teach clinical skills a teacher must learn to involve patients in the educational process. It is through this process that learners acquire the skills of observation, communication, examination and professionalism. Despite the importance of teaching with the patient present, many clinical teachers are hesitant to teach at the bedside. This paper describes a workshop on bedside teaching. The authors present barriers and advantages discussed during the workshop as well as a 'Model of Best Bedside Teaching Practices', which emerged after conducting the workshop for over 135 medical educators. The model includes suggested skills for effective bedside teaching that are arranged into three domains: attending to patient comfort, focused teaching and group dynamics. PMID- 12745519 TI - Evaluating an instrument for the peer review of inpatient teaching. AB - The purpose of this study was to assess an instrument for the peer review of inpatient teaching at Mayo. The Mayo Teaching Evaluation Form (MTEF) is an instrument, based on the Stanford seven-category educational framework, which was developed for the peer review of inpatient teaching. The MTEF has 28 Likert scaled items derived from the Stanford Faculty Development Program form (SFDP 26), the Mayo electronic evaluation form and three additional items. In this study three physician-evaluators used the MTEF to evaluate 10 attending physicians on the Mayo general internal medicine hospital services. Cronbach's alphas were used to assess the internal consistency of the MTEF, and Kendall's coefficient of concordance was used to summarize the inter-rater reliability. Results of this study reveal that the MTEF is internally consistent, based on average ratings across all evaluators (Cronbach's alpha=0.894). Stanford categories with the highest alphas are Self-Directed Learning, Learning Climate, Communication of Goals, and Evaluation. Categories with lower alphas are Feedback, Understanding and Retention, and Control of Teaching Session. Additionally, the majority of items on the MTEF show significant agreement across all evaluators, and teacher enthusiasm was among the most reliable items. In conclusion, the MTEF is overall internally consistent for the peer review of inpatient teaching at Mayo. Hence, the MTEF may be a useful element in the peer evaluation of teaching at our institution. PMID- 12745520 TI - Developing and evaluating ambulatory care: problem-based learning cases. AB - Medical educators have raised concerns about the quality of teaching and learning in busy ambulatory care settings. Problem-based learning (PBL), which allows students to learn to diagnose and manage common ambulatory care problems as they discuss patients away from the clinical setting, is one possible solution for addressing these concerns. This article describes a process for developing realistic and well-written PBL cases for an ambulatory care clerkship. The process details specific steps for writing and evaluating cases to ensure they contain relevant learning issues students often encounter in outpatient training sites. Faculty at other institutions can adapt this process to develop and evaluate PBL cases reflecting the common presenting problems and patient issues at their sites. PMID- 12745521 TI - Training pre-hospital trauma care in low-income countries: the 'Village University' experience. AB - A prospective study was carried out of the health effects and sustainability of a low-cost trauma training program for non-graduate village healthcare workers. From 1996 to 1999, a core group of 44 health workers from mine-infested rural communities in Cambodia and Northern Iraq were trained and equipped to deliver low-cost life support to trauma victims. They in turn trained a network of 2800 layman village first responders. Training was done in makeshift camps at village level ('Village Universities'). A total of 813 patients were managed by the rural rescue system from 1997 to 1999. The mortality rate for trauma victims decreased from 22.6% in 1997 to 13.7% in 1999 (95% CI for difference 1.8% to 16.0%). Management by village first responders had a significant impact on in-field response times and trauma mortality. The rescue system replicated itself during the study period as an indicator of sustainability. The study showed that after trauma care training at rural makeshift training centers, non-graduate health workers can build efficient and sustainable rural rescue systems. PMID- 12745522 TI - Emergency procedure skills of graduating medical doctors. AB - In the present study final-year medical students' degree of theoretical knowledge and rate of successful performance of emergency procedures was assessed. A questionnaire was sent to all final-year medical students in Finland in 1997 (n=504) in all five medical faculties. The response rate was 80.2% (n=404). The questionnaire included questions on theoretical knowledge and successful performance of 10 emergency procedures. Over 90% of the final-year medical students knew the theory of emergency procedures, with the exceptions of chest tube insertion (84%), pericardiocentesis (47%), and planning and starting fluid infusion for an infant (83%), and over 90% had successfully performed insertion of an intravenous line (100%) and intubation of an adult (90%). However, fewer than 7% of the students had successfully performed chest tube insertion, planned fluid infusion for an infant, or pericardiocentesis. Males had significantly higher odds ratio than females for performing insertion of the intravenous line and intubation of an infant. Students with working experience had higher odds ratios for performing cardiopulmonary resuscitation. The highest overall frequency of the procedure performance was at the university in which a student logbook was systematically used. Conclusions are that final-year medical students have good theoretical knowledge of emergency procedures, but practical teaching should be encouraged, since even in emergency procedures students' experience of practical measures was low. PMID- 12745523 TI - Professional socialization revised: medical students' own conceptions related to adoption of the future physician's role--a qualitative study. AB - This qualitative study investigated medical students' experiences of and conceptions concerning their professional development during their first clinical year. Twenty-two medical students participated in a portfolio course. Their personal writings were analysed by qualitative content analysis. Self-image as a future doctor rapidly evolved during patient contacts in the first clinical year. At the beginning, students felt lacking in credibility in front of their patients but towards the end they enjoyed their role as student-physicians. The medical students felt intense stress but the majority of this may stem from strong emotional experiences rather than the medical knowledge to be absorbed. Students were afraid of being humiliated by hospital staff and they felt themselves to be outsiders. No cynicism or suppression of feelings was observed in writings. The first clinical year is a period of intense emotional experiences and rapid development. Portfolios as learning tools may help in recognizing key experiences and in supporting professional development. PMID- 12745524 TI - The development and evaluation of a community attachment scheme for first-year medical students. AB - This paper describes the development over 14 years of a Community Attachment Scheme for First Year Medical Students in Sheffield, together with feedback from tutors and students. The scheme involves pairs of students visiting families expecting a baby or experiencing an illness. The families are identified by general practitioners who act as tutors together with a behavioural scientist for groups of eight to 10 students. The scheme provides first-year students with practical experience of sociology and psychology in terms of family dynamics and illness behaviour. Assessment is part of the degree examination, and involves a written assignment on the family, together with tutors' assessments. The development of the attachment scheme took place in three phases, which are described together with feedback from tutors and students, as well as changes in methods of assessment. The basis of the Community Attachment Scheme has been self directed problem-based learning in small groups with continuous assessment, and these principles have now extended to the rest of the medical curriculum in Sheffield, of which the Community Attachment Scheme is an integral part. PMID- 12745525 TI - Interviewing and case presentations in the presence of patients in undergraduate psychiatry: an experiment. AB - A project to teach psychiatry to medical students in a protected workshop is described. The aim was to provide extended exposure to mental illness in the community but also for the students to present their findings to their peers and teacher in the presence of the patient. From a content analysis of the comments of the students and the patients, it emerges that while the experience was a positive one overall, the students were uncomfortable asking personal questions, and had particular difficulty presenting the patient's mental state and diagnosis in the patient's presence. Many of the patients were ultra-orthodox Jews and the conditions of the interviews often caused them religious concern. The experience was important for its emphasis on communication, and the feedback from the students and patients succeeded in identifying areas for change, such as accommodating religious concerns, and discussing with the patient sensitive issues such as the meaning of a diagnosis of schizophrenia. PMID- 12745526 TI - The CIP (comprehensive integrative puzzle) assessment method. AB - This paper describes a novel tool for assessment in medical education, the comprehensive integrative puzzle (CIP). The dual scoring system of the puzzle stresses the integrative elements of diagnostic thinking and clinical reasoning, while preserving the ability to discern proficiency in various disciplinary elements. The CIP has the format of an 'extended matching' crossword puzzle. Its answer sheet is a grid comprising rows and columns. The left-hand column contains diagnoses or brief clinical vignettes. To complete the cells of the grid the student is required to match, stepwise, the various 'disciplinary investigations' to the diagnoses or clinical vignettes. When the puzzle is completed each horizontal row reflects a coherent medical case. The completed horizontal rows reflect integrative ability (diagnostic thinking and clinical reasoning) and the vertical columns measure the student's proficiency in interpreting medical history data, physical examination findings, laboratory test results, ECG, imaging, special tests, pathology and pharmacology. The CIP has been well accepted by teachers and students during the last seven years at the Bruce Rappaport Faculty of Medicine in Haifa, and it has favorably affected both student assessment and teaching. The reliability of the test and its validity will be reported separately. PMID- 12745527 TI - Understanding our mistakes: a primer on errors in clinical reasoning. AB - Clinical reasoning allows physicians to move from areas of clinical uncertainty to points where the medical literature offers guidance, and is equally important in deducing whether the results of clinical trials are applicable to an individual patient. However, studies in the field of cognitive psychology indicate that the reasoning skills of clinicians are imperfect. Moreover, clinicians may be aware of their mistakes but often do not understand the cognitive processes underlying their errors. Greater understanding of the reasoning process has the potential to improve patient care but independent study of clinical reasoning can be difficult, as the literature is complex and unfamiliar to most physicians. This article provides an introduction to diagnostic reasoning and highlights some of the cognitive factors that lead to errors in clinical problem solving. Clinical scenarios are used to illustrate key points and place the material in a readily accessible framework. PMID- 12745528 TI - The design of distance-learning programmes and the role of content experts in their production. AB - This paper describes an effective and efficient approach to the production of distance-learning materials in which content experts, editors and instructional designers collaborate. The approach is based on the development of a clearly defined agreed educational strategy and the use of a template for the programme. This allows the content experts to assemble the first draft of the programme in an appropriate format, with further revisions being carried out by the educationists in collaboration with the content editor. A task-based approach was adopted in the programme and a two-column layout incorporating nine different types of educational enhancements was used. Useful educational strategies which should be considered in the development of the template for a distance learning programme can be categorized into the following three areas: (1). interactivity enhancements including questions posed, think points for reflection, and case scenarios linking theory to practice; (2). enrichment enhancements including illustrations, readings and quotes from experts; (3). action/practice enhancements highlighting the application of portfolio building--provides useful advice on topics covered as they relate to the AFRC examination, and encourages trainees to apply their knowledge. The use made of the enhancements varies in the different modules. This project was challenging and ambitious but our objectives were met with all 39 authors satisfactorily producing the 42 SELECT Units. PMID- 12745529 TI - Promoting self-directed learning for continuing medical education. AB - Self-directed delivery modes for continuing medical education (CME) are the most effective approaches for improving physician performance. However, instructor directed programs are still the most popular methods used for CME. The purpose of the study was to assess the utilization, preferences and barriers to use of nine different CME delivery methods by physicians. A self-administered survey of all licensed physicians in Nevada was conducted over a three-month period. Results were analyzed using SPSS for windows (version 10). In-person conferences (92%) and journal review (64%) were the most frequently utilized modes of instruction. Rural physicians were more likely to use interactive video. The top three ranked preferences were in-person conference, print-based self-study and CD-ROM. It is concluded that computer training, dedicated time in the workplace for self directed methods, and the development of more interactive CD-ROM and Internet programs will encourage the use of self-directed CME. PMID- 12745530 TI - Involving medical students in staff development activities. AB - Staff development is seen as a key aspect of medical education and many teachers/tutors are now required to attend some formal development activity. However, most workshops do not involve the student. In Manchester, the philosophy is to actively involve students in all aspects of the undergraduate course including delivery and evaluation. The authors have also worked with students in staff development. This predominantly descriptive article sets out some of the ways in which students have been involved in staff development, including co facilitating sessions, role-play and discussion groups. Feedback from 10 workshops was analysed, which showed that 76% of the staff found the student involvement to be the most useful aspect of the workshops, particularly as it allowed staff to engage students in discussion about the course. Both these results and informal feedback from participants suggest that involvement of students is highly valued. Feedback from student volunteers also suggests that they gain from the experience. PMID- 12745531 TI - Determining generic standards for specialist registrar training. AB - Requirements for specialist registrar (SpR) training have been developed by adapting standards already determined for senior house officers in the West Midlands. A consensus of views of chairs of higher specialist training committees on generic requirements for SpR training has been reached by using a Delphi technique. There is a large variation between specialties in the extent that they have been able to reach these standards; some have proved easier to establish than others. Acceptance of the new standards should lead to the delivery of a measurable degree of higher quality education, training and experience for SpRs. PMID- 12745533 TI - Teaching evidence-based medicine to medical students. AB - Given the proliferation of published studies and clinical updates, knowing how to use the literature effectively and efficiently is a necessary skill. Many medical schools in the United States are requiring courses in evidence-based medicine (EBM) in an effort to teach medical students how to distinguish high- from low quality studies, how to interpret results from systematic reviews, and how to recognize flaws in study design or in methodology. The Department of Public Health of the Weill Medical College of Cornell University, in collaboration with the Information Service of the Library, initiated a required EBM course in the first year of medical school. This four-week comprehensive course in the concepts and techniques of EBM focused on methods, study design and statistical analysis in assigned articles as well as on instruction in database-searching techniques. This report reviews the process of implementing such a course and factors needed to ensure the realization of its objectives. PMID- 12745532 TI - The application of a core curriculum and options to an undergraduate paediatric course: a personal view. AB - The time allocated to the undergraduate paediatrics curriculum has been reduced in recent years. It is therefore increasingly important to organize a structured curriculum that provides students with the necessary competences to care for sick children in the early years after graduation. In this paper, the educational strategy of a core curriculum and options is discussed and its application to an undergraduate paediatrics course is described. A core curriculum with options can be readily adapted to undergraduate paediatrics but it requires careful planning to provide the student with the necessary competences and a rewarding and enjoyable educational experience. PMID- 12745534 TI - Computer-assisted learning for teaching anatomy and physiology in subjects allied to medicine. AB - Computer-assisted learning (CAL) may provide an effective supplement to conventional methods of teaching, particularly in subjects such as anatomy and physiology. CAL provides the student with an important additional resource and facilitates alternative modes of learning that are well suited to the requirements of students in subjects allied to medicine. A brief survey is presented which examines those studies that have investigated the utility of CAL for teaching anatomy and physiology within subjects allied to medicine. PMID- 12745535 TI - Experience with a program of faculty development. AB - The authors describe the faculty development program at the University of Nebraska Medical Center. Faculty needs were identified in instructional skill development, academic socialization and mentoring. Committees with campus-wide representation designed the instructional activities. Among the total 749 faculty, 59% attended at least one faculty development offering consisting of one and two-day institutes or two-hour luncheon workshops in the past five years. Evaluations ranked each event highly for quality, relevance, impact on teaching and usefulness. Experiences in creating a successful faculty development program at an academic medical center are reported. Success was measured by attendee numbers and increased participation of faculty in teaching and mentoring. Factors contributing to this success include generous financial support by leadership, broad-based planning and administrative support. PMID- 12745542 TI - Human intestinal epithelial and smooth muscle cells are potent producers of IL-6. AB - BACKGROUND: Interleukin-6 (IL-6), a pluripotent cytokine, has traditionally been considered the product of proinflammatory cells. However, many other cell types have been shown to produce IL-6. Since intestinal inflammation is commonly associated with a vigorous systemic inflammatory response, we hypothesized that intestinal epithelial and smooth muscle cells might contribute to that response by producing IL-6. We therefore studied the capacity of differentiated human intestinal epithelial and smooth muscle cell lines to produce IL-6 in response to various proinflammatory stimuli. MATERIALS AND METHODS: CCL-241, a human intestinal epithelial cell line, and HISM, a human intestinal muscle cell line, were grown to confluency and then treated for 24 h with various concentrations of lipopolysaccharide, Clostridium difficile culture extract containing both toxin A and toxin B, recombinant human tumor necrosis factor-alpha (TNF-alpha), or recombinant human interleukin-1 beta (IL-1beta). Supernatants were then collected for IL-6 determination using an enzyme-linked immunosorbent assay. Cell numbers were determined using a Coulter counter. For comparison, parallel studies were performed using phorbol ester-primed U-937 and THP-1 human macrophage cell lines. RESULTS: Both human intestinal epithelial and smooth muscle cells produced IL-6 under basal conditions. In HISM cells, but not in CCL-241 cells, IL-6 release was increased slightly by treatment with C. difficile culture extract containing both toxin A and toxin B and with lipopolysaccharide. In both cell lines, IL-6 production was profoundly stimulated by treatment with IL-1beta and less so with TNF-alpha. Combinations of high-dose TNF-alpha and IL-1beta may have a slightly additive, but not synergistic, effect on IL-6 release. The amount of IL-6 produced by IL-1-stimulated intestinal cell lines was 70-fold higher than that produced by stimulated macrophage cell lines. CONCLUSIONS; Both intestinal epithelial and smooth muscle cells demonstrate the ability to release significant amounts of IL-6. The profound response to IL-1beta and TNF-alpha stimulation by both cell lines suggests that human intestinal parenchymal cells, influenced by paracrine mediators liberated from proinflammatory cells, might significantly contribute to the overall systemic inflammatory response by producing IL-6. PMID- 12745544 TI - Acute effect of hemodialysis on serum levels of the proinflammatory cytokines. AB - Chronic inflammation is a common feature of end-stage renal disease, which carries a heightened risk of atherosclerosis and other co-morbid conditions. Dialysis treatment per se can bring additional risk factors for inflammation, such as increased risk of local graft and fistula infections, impure dialysate or bio-incompatible membranes. Our study was designed to determine whether a hemodialysis session leads to an acute substantial alteration in the plasma levels of the proinflammatory cytokines interleukin (IL)-6, IL-1beta and tumor necrosis factor (TNF)-alpha, the T-lymphocyte activation factor soluble IL-2 receptor (sIL-2R), and an inflammation mediator and chemotactic granulocyte factor, IL-8, in end-stage renal disease patients receiving chronic intermittent HD. In this study, 21 (12 male/nine female) patients undergoing chronic hemodialysis were enrolled. The acute effect of a hemodialysis session on serum cytokine concentrations was assessed by comparison of pre-hemodialysis and post hemodialysis determinations. Serum IL-1beta, sIL-2R, IL-6, IL-8 and TNF-alpha levels were determined with chemiluminescence enzyme immunometric assays. A significant difference was not observed for IL-1beta, IL-6, TNF-alpha, and sIL-2R concentrations in pre-hemodialysis and post-hemodialysis specimens (p>0.05). Serum median (25th-75th percentiles) IL-8 concentration was 69.4 (34.9-110.3) pg/ml before hemodialysis, and decreased to 31.5 (18.0-78.8) pg/ml following hemodialysis (p: 0.006). Clearance of IL-8 increased by 0.47+/-0.08 pg/ml for each unit increase in pre-dialysis IL-8 (p<0.001) and decreased by 5.63+/-2.59 pg/ml for each unit increase in pre-dialysis urea mmol/l (p<0.05). In conclusion, the results of our study demonstrate that a hemodialysis session markedly decreases IL-8 concentration, which is significantly affected by pre-dialysis concentrations, indicating that removal of IL-8 is a concentration gradient dependent action, but does not change the serum levels of IL-1beta, sIL-2R, IL-6, and TNF-alpha, underlining importance of the structural characteristics of the molecules. PMID- 12745543 TI - Correlation of serum tumor necrosis factor-alpha, interleukin-4 and soluble interleukin-2 receptor levels with radiologic and clinical manifestations in active pulmonary tuberculosis. AB - The precise clinical manifestations of tuberculosis are likely to result from a complex interaction between the host and the pathogen. We took serum samples from a group of patients with a variety of clinical and radiological stages of pulmonary tuberculosis in order to characterize tumor necrosis factor-alpha (TNF alpha), interleukin-4 (IL-4) and soluble interleukin-2 receptor (sIL-2R) response. We further evaluated whether the levels of TNF-alpha, IL-4 and soluble IL-2R are related with each other, and also evaluated the levels of TNF-alpha, IL 4 and sIL-2R after anti-tuberculosis therapy and relation with radiologic scores. Forty-three inpatients with active pulmonary tuberculosis and 19 healthy controls participated in the study. Patients were divided into four categories radiologically on chest X-ray (minimal, moderate-advanced, far-advanced and with miliary infiltration). Concentrations of TNF-alpha (20.9+/-10/15.4+/-8 pg/ml) and sIL-2R (2569+/-842/1444+/-514 pg/ml) were statistically different between patients and controls (p=0.02 and p=0.0001, respectively). Before chemotherapy there was a positive correlation between TNF-alpha and sIL-2R (r=0.34), but there was no correlation between IL-4 and TNF-alpha, and between IL-4 and sIL-2R (r= 0.23 and r=-0.22). The TNF-alpha level was not statistically different in four groups before and after chemotherapy. Results of this study provided some evidence confirming the previously reported role of TNF-alpha, IL-4 and sIL 2R in the control of tuberculosis, but these cytokines were not found related with disease severity. PMID- 12745545 TI - Effect of pharmacologically induced smooth muscle activation on permeability in murine colitis. AB - BACKGROUND: Both intestinal permeability and contractility are altered in inflammatory bowel disease. Little is known about their mutual relation. Therefore, an in vitro organ bath technique was developed to investigate the simultaneous effects of inflammation on permeability and smooth muscle contractility in different segments of the colon. METHODS AND MATERIALS: BALB/c mice were exposed to a 10% dextran sulphate sodium drinking water solution for 7 days to induce a mild colitis, while control mice received normal tap water. Intestinal segments were placed in an oxygenated organ bath containing Krebs buffer. Permeability was measured by the transport of the marker molecules 3H mannitol and 14C-polyethyleneglycol 4000. Contractility was measured through a pressure sensor. Smooth muscle relaxation was obtained by salbutamol and l phenylephrine, whereas contraction was achieved by carbachol and 1-(3 chlorophenyl)-biguanide. RESULTS: The intensity of mucosal inflammation increased throughout the colon. Also, regional differences were observed in intestinal permeability. In both normal and inflamed distal colon segments, permeability was diminished compared with proximal colon segments and the non-inflamed ileum. Permeability in inflamed distal colon segments was significantly decreased compared with normal distal segments. Pharmacologically induced relaxation of smooth muscles did not affect this diminished permeability, although an increased motility positively affected permeability in inflamed and non-inflamed distal colon. CONCLUSIONS: Inflammation and permeability is inversely related. The use of pro-kinetics could counteract this disturbed permeability and, in turn, could regulate the disturbed production of inflammatory mediators. PMID- 12745546 TI - Upregulation of expression of the chemokine receptor CCR5 by hydrogen peroxide in human monocytes. AB - The objective of this study was to test the hypothesis that an oxidative stress can serve as a signal to regulate the expression of CCR5. When human monocytes were exposed to graded concentration of hydrogen peroxide (H(2)O(2)), CCR5 mRNA levels increased maximally at 4 h of exposure to 200 microM of H(2)O(2) and decreased by 24 h of treatment. Pretreatment of monocytes with the NF-kappaB inhibitor BAY 11-8072 blocked the H(2)O(2)-induced augmentation of CCR5 mRNA expression, suggesting a role for this transcription factor in the regulation of CCR5 expression. CCR5 protein expression on the plasma membrane was also increased by treatment with H(2)O(2,) as assessed by flow cytometry. This was accompanied by enhanced responsiveness of H(2)O(2)-pretreated monocytes to the CCR5 ligand MIP-1beta in terms of chemotaxis and c-fos gene activation. Our results suggest that oxidative stress may indeed modulate the expression of chemokine receptors and thus contribute to regulation of the inflammatory process. PMID- 12745549 TI - Surviving hepatic rupture in pregnancy--a literature review with an illustrative case report. AB - Spontaneous hepatic rupture in pregnancy is a rare condition associated with significant maternal and perinatal mortality. Patients developing pre-eclampsia and especially HELLP syndrome require close monitoring for prompt diagnosis of hepatic rupture. However, the presenting symptoms and signs, e.g. epigastric pain, shoulder pain, nausea and vomiting, are common. Thus a high index of suspicion and early evaluation with imaging is vitally important. The important lesson to be learnt is that a Pfannenstiel incision does not allow for adequate assessment of the liver. If hepatic rupture is suspected a second upper abdominal incision should be performed. Improved survival can be achieved through early recognition and a multidisciplinary approach. PMID- 12745548 TI - Exhaled nitric oxide in mylar balloons: influence of storage time, humidity and temperature. AB - BACKGROUND: Mylar balloons are used to collect exhaled air for analysis of fractional nitric oxide concentration (FENO). AIM: We studied the effect of storage conditions on the stability of nitric oxide (NO) in mylar balloons. METHODS: Exhaled air samples and calibration gases were stored in mylar balloons at 4, 21 and 37 degrees C, with or without silica gel. NO was measured after 0, 6, 9, 24 and 48 h. Scheffe F-tests were used to compare NO values. RESULTS: NO remained stable in balloons for 9 h at all temperatures, without silica gel. NO increased between 9 and 48 h, but only with low initial FENO. Silica gel increased variability. CONCLUSIONS: FENO in mylar balloons is stable for at least 9 h. The storage temperature is not critical, but silica gel increases variability. PMID- 12745547 TI - Dexamethasone prevents interleukin-1beta-induced nuclear factor-kappaB activation by upregulating IkappaB-alpha synthesis, in lymphoblastic cells. AB - AIMS: Glucocorticoids (GCs) exert some of their anti-inflammatory actions by preventing the activation of the transcription factor nuclear factor (NF)-kappaB. The GC-dependent inhibition of NF-kappaB may occur at different levels, but the mechanisms involved are still incompletely understood. In this work, we investigated whether the synthetic GC, dexamethasone (Dex), modulates the activity of NF-kappaB in the lymphoblastic CCRF-CEM cell line. We also evaluated the ability of Dex to prevent the activation of NF-kappaB in response to the potent proinflammatory cytokine, interleukin (IL)-1beta. RESULTS: Exposure of the cells to Dex (1 microM) induced the rapid degradation of IkappaB-alpha, leading to the transient translocation of the NF-kappaB family members p65 and p50 from the cytoplasm to the nucleus, as evaluated by western blot. Electrophoretic mobility shift assays revealed that, in the nucleus, these NF-kappaB proteins formed protein-DNA complexes, indicating a transient activation of NF-kappaB. Additionally, Dex also induced de novo synthesis of IkappaB-alpha, following its degradation. Finally, when the cells were exposed to Dex (1 microM) prior to stimulation with IL-1beta (20 ng/ml), Dex was efficient in preventing IL-1beta induced NF-kappaB activation. The GC antagonist, RU 486 (10 microM), did not prevent any of the effects of Dex reported here. CONCLUSION: Our results indicate that, in CCRF-CEM cells, Dex prevents NF-kappaB activation, induced by IL-1beta, by a mechanism that involves the upregulation of IkappaB-alpha synthesis, and that depends on the early and transient activation of NF-kappaB. PMID- 12745550 TI - Retinal detachment and the second stage of labour: a survey of regional practice and literature review. AB - Following several referrals for 'assessment', obstetricians in the West Midlands Health Region were surveyed regarding patients perceived to be at high risk of retinal detachment during the second stage of labour. A questionnaire was sent to 95 practicing obstetricians. This was a four-item questionnaire, based on four identifiable categories of referral to the ophthalmologist, by the obstetricians. It also covered the perceptions of the obstetricians of the ocular problem in relation to their obstetric clinical decision. With a 70% response rate, a significant number of obstetricians consider that certain ophthalmic conditions predispose to retinal detachment during labour. Even though they hold this view, the obstetricians generally do not modify their obstetric practice in response to any perceived ophthalmic risk. Such a view is not borne out by current literature, with no evidence to support a causative link between labour and rhegmatogenous (versus serous) retinal detachment, even in eyes predisposed to that condition. PMID- 12745551 TI - Active management of labour revisited: the first 1000 primiparous labours in 2000. AB - This was a prospective observational study in a tertiary referral obstetric unit in Dublin to ascertain the delivery outcome in the first 1000 nulliparous women in 2000. The initial diagnosis of labour was confirmed in the first 1000 consecutive nulliparous women in spontaneous labour with cephalic presentation at term. All patients underwent active management of labour. Active management included strict criteria for the diagnosis of labour, early amniotomy, 2-hourly vaginal examinations, oxytocin augmentation where progress of labour was slow and the presence of a companion (personal nurse) in labour. Epidural analgesia was freely available. Mode of delivery, duration of labour, analgesia usage and maternal and perinatal complications were the main outcome measures. All patients presented with painful uterine contractions, 75% with show in addition, and 36% had spontaneous rupture of membranes on admission. Eighty per cent presented with a cervical dilatation of 12 hours) was 4.3%. Postpartum haemorrhage occurred in 3.8% of mothers and 1.6% of babies were admitted to the special care baby unit. Our study suggests that active management of labour is associated with a low incidence of prolonged labour and a low caesarean section rate. PMID- 12745552 TI - Evaluation of an experimental midwife-led unit in Scotland. AB - This paper reports on the outcome of pregnancy among 3322 low-risk women managed in an 'experimental midwife-led unit' at Forth Park Hospital, Kirkcaldy, Scotland. All women were allocated to receive midwife-led antenatal and intrapartum care. Of the 3322 women, 1786 were nulliparous and 1536 were parous. Of the 1786 nulliparous women, 532 (30%) were transferred to consultant care antepartum, 488 (27%) were transferred intrapartum and the remaining 766 (43%) were delivered in the midwife-led unit (MLU). Of the 1536 multiparous women, 343 (22%) were transferred to consultant care antepartum, 158 (10%) were transferred intrapartum and the remaining 1035 (68%) were delivered in the MLU. Among the intrapartum transfer group, 14% delivered during the first hour after the decision to transfer and another 14% gave birth during the second hour (28% between 0-2 hours). More than half of the women transferred during the second stage delivered within the first hour. Of the 2447 admitted to the MLU, 32% were seen by obstetric team on more than one occasion. Of the 1801 babies delivered in the MLU, 9% required resuscitation and 2% of all the babies were admitted to the special care baby unit. This study suggests that present antenatal criteria are unable to determine who will remain at low risk throughout pregnancy and labour, especially among nulliparous women. The clinical situation could change adversely during labour, requiring the involvement of medical staff. Commissioners of maternity services may wish to utilise these data to formulate local protocols for stand-alone midwife-led units. PMID- 12745553 TI - Diagnostic features of cervical incompetence among women in Maiduguri. AB - The study was a review of the clinical features used to diagnose cervical incompetence. One hundred and forty-one clinical records of patients who had cervical cerclage inserted for cervical incompetence at the University of Maiduguri Teaching Hospital over a 5-year period, were studied retrospectively. Previous mid-trimester abortion occurred in 80.1% of the patients and was consecutive in 98 patients (69.5%). Sixty-eight (50.7%) patients had previous preterm deliveries, while 65 (46.1%) had a cervical cerclage inserted in a previous pregnancy. They had associated characteristic rapid painless expulsions of the fetus in previous pregnancies. One hundred and thirty-six patients had the McDonald's method of insertion (96.5%). There was a significant association between histories of previous midtrimester abortions, consecutive mid-trimester abortions and preterm delivery, with a significantly greater proportion of women with a previous history of preterm delivery having a recurrence in spite of the insertion of the cervical cerclage. PMID- 12745554 TI - The role of androgens in the aetiology and pathology of pre-eclampsia. AB - Pre-eclampsia and eclampsia, along with its complications such as prematurity, intrauterine growth retardation, perinatal asphyxia and abruptio placenta, continues to be one of the major causes of maternal and fetal morbidity and mortality. Due to its morbid course, it is necessary to identify those at risk for the illness and take precautions. However, the lack of knowledge about the aetiology makes it difficult to assess risk factors. We studied the role of the serum androgens in the pathogenesis of pre-eclampsia. Ninety severe pre eclamptic, 10 mild pre-eclamptic and 50 normotensive pregnant women were studied. The patient and control groups' blood total testosterone, free oestriol (FE3), FAI (free androgene index), sex hormone binding globulin (SHBG) and dehydroepiandrostenedione sulphate (DHEA-S) values were compared. The sex, weight, APGAR scores and hospitalisation in the neonatal intensive care unit of the babies were also evaluated. No statistically significant difference in the total testosterone and free oestriol values was found between the groups. SHBG was found to be higher in the pre-eclamptic group, whereas free testosterone and DHEA-S were higher in the normotensive group. Compared to the severe pre eclamptic group, no difference was found in the total testosterone levels, whereas free testosterone levels were significantly higher in the mild pre eclamptic group. We reached the opinion that androgens do not play a clinically significant role in the pathogenesis of pre-eclampsia. PMID- 12745555 TI - Current practice for induction of labour in the United Kingdom: time for a review? AB - This paper presents the results of postal survey of protocols for induction of labour. The drug regimes used for the induction of labour in NHS hospitals in England and Wales have been compared to both RCOG guidelines and to the drug manufacturer's recommendations. A wide variation in practice was found, with many units using dosages that are different from those recommended. The implications of such departures from the norm are discussed in terms of the importance of evidence-based practice and the role of national guidelines in ensuring optimum care. PMID- 12745556 TI - Female genital mutilation and the unborn female child in southwest Nigeria. AB - Female genital mutilation, despite efforts to abolish it, is still widely practised in Nigeria. The risk of female genital mutilation to a female child in southwest Nigeria was investigated by interviewing 430 consecutive pregnant women attending the antenatal clinic of Wesley Guild Hospital Ilesa, Nigeria between July 2001 to October 2001. The results show that 60% of the pregnant women studied had a type of genital mutilation. The decision to mutilate a female child is taken before she is born. Seventy-four (17.2%) of the women and 146 (34%) of their husbands would circumcise their female child. The decision to circumcise a female child is made between the husband and wife but the final decision comes mainly from the husband. Because the majority of the women (58.4%) were yet to decide whether or not to circumcise their female children, they could sway the decision either way before the husband makes up his mind. Therefore, every effort should be taken to involve men in the struggle to eradicate this unwholesome practice. PMID- 12745557 TI - The changing trend in the rate of caesarean section at a teaching hospital. AB - This is a retrospective study analysing the reasons behind the observed increasing rate of caesarean section (C/S) over a 10-year period (1990-99) in the obstetric unit of Jordan University Hospital. All the indications of C/S carried out in the unit were analysed. The duration of the study was divided into 2, of 5 years each (1990-94, 1995-99). There was a 6.9% increase in the C/S rate over the second half of the study period. This was statistically significant (P < 0.001). All the indications contributed significantly to rise. Fetal distress had the highest contribution 33.5%, while repeat C/S and malpresentation contributed to 21.5% and 21.3%, respectively. This increase was not associated with a significant change in the perinatal mortality. The rise in the caesarean section rate was higher in primigravida compared with multigravida (10.9% vs 6.2%). Fetal distress has the highest contribution in primigravida. In multigravida, if we exclude repeat caesarean section, the major indications were fetal distress and malpresentation. The percentage of elective and emergency caesarean section was similar in both study periods. The reasons behind the increase C/S rate couldn't be understood. Probably a lower threshold concerning the decision to perform the C/S rather than a change in obstetric management in responsible for this rise. PMID- 12745558 TI - A randomised, double-blind placebo-controlled trial of ascorbic acid supplementation for the prevention of preterm labour. AB - In a previous study from this institution, patients at high risk for preterm labour were screened for the presence of bacterial vaginosis (BV). When BV was present, they were randomised to receive either treatment (metronidazole) or placebo (vitamin C). There were significantly more patients with preterm labour in the metronidazole group. The aim of this double-blind randomised placebo controlled trial study was to determine whether vitamin C could indeed reduce the recurrence risk of preterm labour. Patients with a history of preterm labour in a preceding pregnancy were randomised to receive 250 mg vitamin C or a matching placebo twice daily until 34 weeks' gestation. They attended a dedicated premature labour clinic. Significantly more women delivered before term in the group that received vitamin C, but there was no difference in the outcome of the babies between the two groups. Supplementation with vitamin C did not prevent premature labour. PMID- 12745559 TI - Goitre prevalence in pregnant women attending antenatal clinic in a teaching hospital. AB - This study aimed to determine goitre prevalence in pregnant women. Seven hundred pregnant women attending the antenatal clinic at Wesley Guild Hospital, Ilesa, were interviewed and examined. The mean age of the subjects was 27.8 years, with mean parity of 2.65. Mean gestation was 28.5 weeks. A total of 97.4% were resident in Ilesa and surrounding towns and villages. Well or stream was the main water supply for nearly all the subjects. Only 7.3% of the subjects showed no thyroid enlargement. Early enlargement (1(a) and 1(b)) occurred in 441 (63%) patients, Grade 2 enlargement in 166 (23.2%) and a grossly enlarged goitre in 40 (5.7%). The goitre rate was correlated with the age and parity of the patients. PMID- 12745560 TI - Screening for gestational diabetes: a retrospective audit. AB - There are various indications for screening pregnant women for gestational diabetes. Screening is then often carried out by performing a timed random blood sugar (t-RBS). In this unit a raised t-RBS is an indication to perform a glucose tolerance test (GTT), and if this is abnormal, the patient is managed with dietary modification or insulin therapy. A t-RBS is considered abnormal if the fasting value (> 2 hours since last meal) is > 5.7 mmol/l, or the non-fasting value (<2 hours since last metal) is > 6.3 mmol/l. However, higher t-RBS values have been recommended by the Diabetic Pregnancy Study Group. This group considers a t-RBS abnormal if the fasting value is > 6.1 mmol/l, or the non-fasting value is > 7.0 mmol/l. A retrospective audit was carried out to determine if cases of gestational diabetes would have been missed if the new guidelines were adopted. There were 112 patients with a fasting t-RBS of 5.8 to 6.1 mmol/l, inclusive, and 196 patients with a non-fasting t-RBS of 6.4 to 7.0 mmol/l, inclusive; 45 patients did not have a GTT result. Therefore 263 patients were included in the study. The number of patients with a normal GTT result was 257 (97.7%), and 6 (2.30%) patients had an abnormal GTT result. Only 4 (1.52%) were labelled as gestational diabetics, and all these cases were managed with diet alone with no adverse obstetric outcome. A large number of GTTs are performed unnecessarily in our unit, and the values recommended by the Diabetic Pregnancy Study Group should be adopted. PMID- 12745561 TI - Adolescent maternal mortality in Sokoto, Nigeria. AB - The aim of this study was to determine the magnitude of adolescent maternal deaths at the Usmanu Danfodiyo University Teaching Hospital (UDUTH), Sokoto. We also studied the associated and causative factors, and recommended strategies for their prevention. This was a retrospective, cross-sectional study of adolescent maternal deaths that occurred at UDUTH, Sokoto from January 1990 to December 1999. There were 946 live births (LB) from adolescents and 46 maternal deaths during the study period. The maternal mortality ratio (MMR) in adolescents was 4863/100000 LB and 2151/100000 LB in the general hospital population. The mean age of the adolescents who died was 17 years. Risk factors included absence of routine antepartum, intrapartum and postpartum care, illiteracy and poverty. Eclampsia and prolonged obstructed labour were responsible for 76% of deaths. Abortion was an uncommon cause of maternal death in adolescents. There is an unacceptably high level of maternal mortality in adolescents at UDUTH, Sokoto. Tragically, most deaths are preventable. PMID- 12745562 TI - Pregnancy outcomes among nulliparous teenagers in suburban Nigeria. AB - Our objective was to study the pregnancy outcomes among teenagers in a rural Nigerian setting. Single births to 1394 nulliparous mothers aged 3 years. Confirmatory HLA typing, CFU and viability testing was successfully performed from the same segment as part of a strategy for product release evaluation. When comparing the segment with its corresponding CB unit, the total colony-forming units (CFU) measured in the two was similar (P = 0.51, paired t test). Three research units purposely sabotaged by an overnight thaw and refreeze had no CFU growth, but viability as measured by Trypan was still 68-98%. DISCUSSION: No deterioration of hematopoietic potential has been detected with storage up to 5 years. The contiguous segment CFU is representative of the product, and thus is a useful tool for quality control and confirmation of product viability. Viability, as measured by Trypan blue dye exclusion may be falsely reassuring. PMID- 12745583 TI - Quantifying levels of transplanted murine and human mesenchymal stem cells in vivo by real-time PCR. AB - BACKGROUND: Previously, we demonstrated that murine mesenchymal stem cells (MSCs) injected intracranially into mice expand throughout the central nervous system (CNS). This paper describes real-time PCR (RT-PCR) assays that enables accurate quantification of transplanted cells in vivo. METHODS: RT-PCR assays that amplify sequences in the mouse Y chromosome or human Alu repeats were developed and used to quantify the number of male, murine, or human MSCs in the CNS at various times after intracranial injection into neonatal mice, or in various organs of adult mice after i.p. injection of cells into 3 day-old embryos. RESULTS: In the CNS, levels of male mouse DNA in female transplant recipients increased on average 30 fold between 3 and 60 days post-injection but then was unchanged at 140 days post transplant (P = 0.107). Male DNA accounted for up to 0.309% of the total DNA content of the brain, representing maximally 600000 donor cells. Human DNA was detected in the CNS up to 300 days post-transplant, but levels never exceeded 7.63 x 10 (-4) % of the total brain content. After in utero transplantation, human DNA levels ranged from 0.36 x 10(-5) % to 2.14 x 10 (-5) % of the total DNA content of liver, kidney and spleen. significantly higher levels were found in heart (P = 0.06), femur and brain (P = 0.0025). DISCUSSION: RT-PCR assays were developed to quantify levels of male, murine and human cells in vivo following sex-mismatched or xeno-transplants. Due to their accuracy, precision, and sensitivity, these assays provide a versatile alternative to measuring stem-cell engraftment in vivo. PMID- 12745584 TI - Phenotypic analysis of human BM T-cell depleted by soybean lectin agglutination and E rosetting with sheep RBC: relative enrichment of NK cells and a CD3(+),CD2( dim) population. AB - BACKGROUND: T-cell depletion (TCD) of BM allows transplantation across HLA barriers. Although different methods are used throughout the world, the optimal application of TCD still remains unclear, partly due to the lack of thorough analyses of the cellular fractions eliminated or retained in each method, and their possible implications regarding GvHD, GvL, or engraftment. We have analyzed the phenotype of the successive fractions of 19 BM samples depleted by soybean lectin agglutination and sheep erythrocyte rosetting (elimination of T cells that form rosettes through CD2), focusing on the final fraction infused to patients. METHODS: Analysis was performed using three-color flow cytometry and strategies for optimal staining and individualism of the subsets of interest. RESULTS: The relative composition of the lymphoid population varied significantly along the successive steps in TCD: at the agglutination step, B cells and CD4 T cells are greatly reduced, while natural killer cells (NK) and TCRgammadelta+ T are augmented. The rosetting steps imply the relative enrichment of CD2-dim T cells, together with a further rise in the proportion of NK and double-negative T cells frequently TCRgammadelta+. DISCUSSION: The presence of minor subsets of CD2- TCRgammadelta+ and CD2- TCRalphabeta T cells has already been described in the peripheral blood of normal individuals. We report that, by using this method of TCD, CD2-dim T cells, frequently TCRgammadelta+, are retained in the grafts and infused in patients, together with NK cells as the main lymphoid population. We discuss the possible implications of these populations in the biology of the graft, regarding GvHD, GvL and engraftment. PMID- 12745585 TI - Ex vivo expansion of neutrophil precursor cells from fresh and cryopreserved cord blood cells. AB - BACKGROUND: Neutropenia following cord blood (CB) transplantation may be abrogated by infusion of granulopoietic progenitor cells. The purpose of this study was to determine whether myeloid progenitors can be obtained by ex vivo expansion of cryopreserved cord blood aliquots, and whether these progenitors present the morphologic, biologic and functional properties of myeloid progenitors at various stages of differentiation. METHODS: The cells, plated for 7 days in serum-free medium with SCF, IL-3, G-CSF, Flt3-ligand and thrombopoietin in various combinations were assessed for the expression of CD34, CD38 and CD13. Maturation of cells into the myeloid lineage was evaluated by the expression of CD15, CD11b and CD16 and by the presence of primary (myeloperoxidase) and secondary granules (lactoferrin). The capacity of cells to phagocyte latex beads was evaluated to assess their functionality. RESULTS: We have shown that a). CD34+ cells isolated from thawed samples were able to produce expansions similar to fresh samples. b). The best combination for the expansion of neutrophil precursor cells was S3FG; c). in these conditions, all stages of myeloid progenitors were represented, but few mature cells were observed. d). However, when the cells were plated on a BM stroma to try to reproduce conditions occurring during transplant, they acquired rapidly the characteristics of mature segmented cells. e). The ex vivo generated granulocytes were able to phagocyte latex beads. DISCUSSION: In conclusion, it seems reasonable to systematically aliquot CB samples before cryopreservation. Some aliquots can then be thawed, enriched in CD34+ cells and ex vivo differentiated into myeloid lineage, while the other aliquots are conserved to be infused without manipulation. PMID- 12745587 TI - Adoptive cellular immunotherapy for non-small cell lung cancer: a pilot study. AB - BACKGROUND: Priming with autologous tumor vaccine followed by ex vivo expansion of activated T cells is a feasible experimental strategy. This paper describes the application of this cellular therapy to treat patients with late-stage non small-cell lung cancer (NSCLC). METHODS: Twenty-one patients with Stages III and IV NSCLC were treated. Tumor samples were obtained surgically (five patients) or by using aco-axial biopsy needle (16 patients). Each course of vaccination consisted of irradiated tumor cells, mixed with GM-CSF and injected intradermally on Day 1, followed by GM-CSF only on Days 2- 5. The course was repeated 10-14 days later. Lymphocytes were collected 10-14 day after the second course and ex vivo expanded using IL-2 and anti-CD3 Ab. The expansion products were then re infused into the patients. RESULTS: Twelve out of 16 biopsies resulted in optimal cell numbers for vaccine preparation. Nineteen out of 21 patients achieved a delayed type hypersensitivity (DTH response after two courses of vaccination. In 18/21 patients, the ex vivo expansion products contained > 1.6 x 10(10) cells. Subset analysis showed 77.0-97.2% T cells with a CD4:CD8 ratio of 0.65-4.0; natural killer cells were 2.0-18.6%. There were no significant toxicities. The median survival of all 21 patients was 18.6 months, with a 1-year survival of 51.6%. CONCLUSION: Autologous tumor cell-vaccination may be combined with ex vivo expansion of lymphocytes as adoptive cellular immunotherapy for advanced NSCLC. Overall survival in this cohort of poor prognosis patients compared favorably with results reported in the literature. PMID- 12745586 TI - Efficacy of granulocyte transfusions for neutropenia-related infections: retrospective analysis of predictive factors. AB - BACKGROUND: The transfusion of G-CSf-primed granulocytes (GTX) might represent an important treatment option for neutropenia-related infections unresponsive to conventional antimicrobial therapies and to recombinant hematopoietic growth factors. However, few studies to date have identified the factors that can predict clinical outcome and the patient populations who are likely to benefit most from GTX. The primary endpoint of the present retrospective study was to evaluate the efficacy of GTX in 22 patients with hematological malignancies who developed neutropenia-related bacterial and fungal infections that were unresponsive to appropriate antimicrobial therapies. METHODS: Peripheral blood granulocytes were collected by continuous-flow leukapheresis from HLA-identical siblings after priming with G-CSF. The response to GTX was classified as 'favorable' if clinical symptoms and signs of infection resolved or 'unfavorable' if clinical symptoms and signs of infection were unchanged or worsened. Control of infection at Day 30 after the enrollment in the GTX program was considered as the outcome variable in multiple regression analysis. RESULTS: Two patients died of infection before receiving the granulocyte concentrates. Bacterial infections (monomicrobial or mixed bacteremias) were documented in 11 patients, whereas fungal infections (fungemia or focal fungal infections) were diagnosed in seven patients. In two patients, no infecting agent could be isolated (clinical infection). Control of infection at Day 30 after the first GTX was achieved in 10 of 20 assemble patients. Overall, 54% of patients with bacterial infections had a favorable response, compared with 57% of patients with fungal infections. No differences in terms of survival were found when comparing patients with bacterial and those with fungal infections at a median follow-up 90 days from the first GTX. In univariate analysis, disease status before GTX, e.g., complete or partial remission, and spontaneous recovery of the neutrophil count were significantly associated with control of infection. when multivariate regression models were formed, the recovery 0.5 x 10 (9)/L PMN was the only parameter that significantly and independently correlated with a favorable response to GTX. DISCUSSION: GTX can be used to successfully treat bacterial as well as fungal infections in severely neutropenic patients when administered early after the onset of febrile neutropenia in patients with remission of the underlying disease and who are likely to recover marrow function. PMID- 12745589 TI - Effects of leukapheresis protocol, cell processing and cryopreservation on the generation of monocyte-derived DC for immune therapy. AB - BACKGROUND: Many clinical trials of DC-based immunotherapy involve administration of monocyte-derived DCs (Mo-DC) on multiple occasion. We aimed to determine tbe optimal cell processing procedures and timing (leukapheresis, RBC depletion and cryopreservation) for generation of Mo-DC for clinical purposes. METHODS: Leukapheresis was undertaken using a COBE Spectra. Two instrument settings were compared - the standard semi-automated software (Version 4.7) (n = 10) and the fully automated software (Version 6.0) (N = 40). Density gradient centrifugation using Ficoll, Percoll, a combination of these methods or neither for RBC depletion were compared. Outcomes (including cell yield and purity) were compared for cryopreserved unmanipulated monocytes and cryopreserved Mo-DC. RESULTS: Software Version 6.0 provided significantly better enrichment for monocytes (P < 0.05) but 25% fewer total monocytes. Final Mo-DC purity was not influenced by leukapheresis or RBC depletion method, but was critically dependent on monocyte adherence. Version 6.0 produced significantly lower RBC and platelet contamination (P < 0.0005) but in vitro RBC depletion could not routinely be omitted. Only 5-6% of monocytes harvested resulted in Mo-DC (95% lost in cell processing or failing to differentiate). DISCUSSION: Cell losses remained significant despite attempts to minimise processing steps during Mo-DC generation. Reduction in RBC and platelets achieved with software version 6.0 was insufficient to offset the disadvantage of the lower monocyte yield. Substantial savings in materials and other costs can be achieved if Mo-DC for multiple treatments are generated from cryopreserved monocytes rather than from fresh monocytes. PMID- 12745588 TI - Functional and immunophenotypic characteristics of isolated CD105(+) and fibroblast(+) stromal cells from AML: implications for their plasticity along endothelial lineage. AB - BACKGROUND: In vitro cultures of BM cells from newly diagnosed patients with AML displayed a defective BM stromal compartment, with a reduced number of fibroblast colony-forming unit (CFU-F: 1 +/- 1.25 SD) and a decreased proliferative ability. The purposes of our study were: 1). to select BM mesenchymal stem cells (MSC) and BM-derived stromal cells (BMDSCs) from AML patients at diagnosis and from healthy subjects, using an immunomagnetic system and either anti-CD105 or anti-fibroblast MAbs; 2). to study the immunophenotypic and functional properties of freshly isolated and cultured mesenchymal cells; 3). to test the in vitro plasticity of the selected cells to differentiate towards an endothelial phenotype. METHODS: Fresh mononuclear cells obtained from BM of 20 patients newly diagnosed with AML and from eight healthy subjects were selected by using anti-fibroblast and anti CD105 MAbs. Freshly isolated cells were analyzed, characterized by flow cytometry using a wide panel of MAbs and seeded in long-term culture medium to assess CFU-F formation. The level of confluence after 30 days and functional capacity in a long-term colony-forming cell culture (LTC-CFC) were tested. Furthermore, the cultured selected cell populations were assayed for their ability to differentiate into an endothelial-like cell phenotype with the addition of vascular endothelial growth factor (VEFG) and endothelial cell growth supplement (ECGS). RESULTS: In normal subjects the selection produced an increase of the CFU F number of 2.6-fold with anti-fibroblast MAb and 2.7-fold with the anti-CD105 MAb. Anti-fibroblast and anti-CD105 MAb selection from AML BM cells resulted in a statistically significant greater count of CFU-F that was respectively 10.6-fold (P = 0.04) and 14.4-fold (P = 0.00001) higher in comparison with the unselected AML samples. Interestingly, in 80% of AML samples immunoselection was also able to restore the capacity of the CFU-F to proliferate and form confluent stromal layers. The isolation of those layers sustained the proliferation and differentiation of hematopoietic stem cells in the LTC-CFC. The phenotypic profile of cultured BMDSCs was different from that of the freshly isolated cells, and changed in relation to the culture conditions: CD105+ selected cells cultured with VEGF and ECGS expressed endothelial markers, a finding that suggests that this cell subpopulation may have the potential to differentiate toward an endothelial-like phenotype. DISCUSSION: We report that immunomagnetic selection represents a valid tool for the selection of BM mesenchymal cells in samples obtained from both healthy subjects and patients with AML. This technique was able to rescue two functional and immunophenotypic compartments related to two different selected populations. In particular, the CD105+ cells isolated in AML displayed, after stimulation with VEGF and ECGS, the ability to change towards an endothelial-like cell phenotype, thus revealing an unexpected plasticity. Both CD105+ and fibroblast+ cells once successfully isolated might represent sources of mesenchymal cells populations useful for in vitro investigations and, above all, as therapeutic devices. PMID- 12745590 TI - Cytokine flow cytometry: multiparametric approach to immune function analysis. AB - More precise quantitation of cellular immune responses has become possible with the advent of single-cell assays of immune function, such as cytokine flow cytometry, enzyme-linked immunospot (ELISPOT), and MHC-peptide multimers. Cytokine flow cytometry is an attractive technique because it allows the detection of responses to whole antigens without regard to MHC restriction, while also collecting additional information on responding cells via multiparameter flow cytometry. In this review, we compare cytokine flow cytometry with other assays of immune function, summarize some of that data that have been collected in various disease states using cytokine flow cytometry, and describe some methodological improvements designed to increase the robustness, throughput, and information content of this technique. We hypothesize that a new generation of automated cytokine flow cytometry assays will allow elucidation of the correlates of protection for diseases involving cellular immunity, through application of these assays in more and large clinical trials. PMID- 12745591 TI - Validation of the single-platform ISHAGE method for CD34(+) hematopoietic stem and progenitor cell enumeration in an international multicenter study. AB - BACKGROUND: Flow cytometric enumeration of CD34+ hematopoietic sterm and progenitor cells (HPC) is the reference point for undertaking apheresis and evaluation of adequacy for PBSC engraftment. An external quality assurance (EQA) scheme for CD34+ HPC enumeration has been operational in Belgium, Netherlands and Luxemburg (Benelux) since 1995. Within this group, a multicenter survey was held to validate the state-of-the-art methodology, i.e., multiparametric definition of HPC based on light scatter, expression of CD34 and CD45, and counting beads (i.e., 'single platform ISHAGE' method). METHODS: 'Real-time' EQA was used to monitor the application of the single-platform ISHAGE method by 36 participants. Three send-outs of stabilized blood with CD34+ cell counts 35-60 cells/microl were distributed to 36 participants, who were required to assay the samples on three occasions using the standard assay and their local techniques. These results were compared with thosed obtained by 111-116 UK NEQAS participants testing the same specimens. RESULTS: Using the single platform ISHAGE methods, between-laboratory coefficients of variations (CVs) as low as 10% were achieved. Intra-laboratory CVs were < 5% for approximately 50% of the participants. Local single-platform techniques yielded between-laboratory CVs as low as 9% in both Benelux and UK NEQAS cohorts. In contrast, the lowest between-laboratory CVs using dual-platform techniques were 17% (Benelux) and 21% (UK NEQAS), respectively. CONCLUSION: The single-platform ISHAGE method for CD34+ cell enumeration has been validated by an international group of 36 laboratories. The observed varation between laboratories allows a meaningful comparison of CD34+ cell enumeration. PMID- 12745594 TI - Intense pulsed light source for the treatment of dye laser resistant port-wine stains. AB - OBJECTIVE: The purpose of this clinical study was to evaluate the efficacy of an intense pulsed light (IPL) irradiator system for the treatment of port-wine stains (PWS) resistant to multiple pulsed dye laser (PDL) treatments. MATERIALS AND METHODS: Fifteen PWS patients, who were previously found to be resistant to multiple PDL treatments, were treated four times with a second generation IPL system. The clinical efficacy was evaluated on close-up photographs 2 months after the last treatment. RESULTS: Patients with dye laser resistant PWS could be divided into two groups: responders to IPL treatments (46.7%) and non responders(53.3%). All responders obtained more than 50% reduction, and 85.7% of the responders obtained between 75% and 100% reduction of their lesions. The group of non-responders was defined as patients who obtained less than 25% clearance. CONCLUSIONS: Approximately half of a group of PWS patients, who did not respond to previous PDL treatments, obtained good or excellent clinical effect after four treatments with the IPL system. All PWS, except those located in the V2 area of the face, responded to the treatments. The IPL treatment modality was found to be safe and efficient for the treatment of PWS, except for those located in the V2 area. PMID- 12745595 TI - Hair removal evaluated with a filterless flashlamp-based system: a preliminary study in 10 patients. AB - BACKGROUND: A relatively inexpensive, portable epilation system based on unfiltered flashlamp technology (Spa Touch, Epilast, Paris, France)was macroscopically and histologically evaluated for efficacy and duration of hair removal. PATIENTS AND METHODS: Ten patients participated (eight female, two male, aged 22-62 years, skin types I-V). The system delivers a 35 ms pulse of 6-7.5 J/cm2 of broadband infrared visible light through a large treatment window in the handpiece, held in light contact with the target skin,without skin cooling or anaesthesia. The skin was shaved before the first treatment, and the subsequent frequency of application over a further four sessions was done every month. RESULTS: Histology revealed the destruction of most of the hair shaft, with regrowth of finer and lighter hair approximately 4 weeks after the final treatment, depending on the area treated. Patients were satisfied with results if epilation was maintained on a monthly basis. A delay in treatment allows hair to grow back. An interesting observation was recorded on white and vellous hair. Nine of the 10 patients noted better skin condition in the treated area. CONCLUSIONS: This system retards hair regrowth and acts as a 'light razor' that can be implemented for speedy epilation at a reasonable cost. PMID- 12745596 TI - Reversal of photodamage with topical growth factors: a pilot study. AB - BACKGROUND: Interest in the reversal of facial photodamage has increased significantly among patients and physicians in the past decade. Though surgical procedures may be very effective, the associated healing time and potential risks have spurred the development of non-surgical treatments. There has also been an increasing depth of knowledge regarding wound healing and its control by growth factors as well as its modulation by the topical application of growth factors. Bioengineered tissue cultures have resulted in the ability to collect naturally occurring human growth factors in their tissue concentrations. OBJECTIVE: The objective of this study is to determine if the twice daily application of a combination of multiple growth factors to photodamaged facial skin results in any evidence of improvement after 60 days. METHODS: Fourteen patients applied a gel containing a mixture of eight different growth factors (Nouricel-MD) to photodamaged facial skin twice daily. Prior to the study and at days there were clinical evaluations of photodamage (Fitzpatrick scale), 3 mm punch biopsies and optical profilometry. Patient questionnaires were answered at 60 days. RESULTS: Eleven of 14 patients showed clinical improvement in at least one facial area. The peri-orbital region showed a statistically significant improvement (p = 0.0003). Optical profilometry showed a statistically significant reduction in Ra measurement (p=0.0075) and shadowing (p=0.02), both indicating a decrease in the depth and number of textural irregularities or fine lines. Biopsies revealed new collagen formation in the Grenz zone (37% increase in thickness) and thickening of the epidermis by 27%. Eight of 14 patients felt their wrinkles were improved, while 12 of 14 felt their skin texture was improved. CONCLUSIONS: The application of a mixture of topical growth factors may stimulate the repair of facial photodamage resulting in new collagen formation, epidermal thickening and the clinical appearance of smoother skin with less visible wrinkling. PMID- 12745597 TI - Er:YAG laser followed by topical podophyllotoxin for hard-to-treat palmoplantar warts. AB - BACKGROUND AND OBJECTIVES: Palmoplantar warts are often hard to treat. They tend to relapse and the course of therapy is frustrating in many cases. The erbium:YAG laser(Er:YAG) with a wavelength of 2.94 rm is capable of achieving a rapid and precise ablation of warts, but about 14% of patients are non-responders as shown in a previous study. Podophyllotoxin is an established antimitotic agent derived from podophyllum plant resin, approved for human papilloma virus (HPV)-induced genital warts. The combination of both ablative Er:YAG laser and topical 0.5% podophyllotoxin solution in hard-to-treat palmoplantar HPV warts was investigated. PATIENTS AND METHODS: Thirty-five patients with hard-to-treat warts(palmar or plantar) with a mean age of (32.2+12.1) years, range 17-50 years, with various pretreatments that had failed, were treated once by Er:YAG laser ablation with a spot size of 3 mm, a frequency between 8 Hz and 10 Hz, and a fluence of 5.7-11.3 J/cm2. After wound healing, topical podophyllotoxin 0.5% solution was applied for 3 days followed by a break of 4 days. Four to six treatment cycles with podophyllotoxin were performed. RESULTS: After laser treatment followed by topical podophyllotoxin cream a complete response was observed in 31 patients (88.6%). Two patients with plantar warts and a complete response showed a relapse within 3 months after treatment (5.7%). None of the patients developed pigmentary changes,wound infections or scarring. CONCLUSION: The therapy of hard-to-treat warts with a combination of Er:YAG laser and topical podophyllotoxin is safe and effective. Compared with laser alone, the CR percentage seems to be higher and the percentage of relapses reduced. PMID- 12745598 TI - Single-pass CO2 laser skin resurfacing of light and dark skin: extended experience with 52 patients. AB - BACKGROUND: Multiple-pass carbon dioxide (CO2) laser skin resurfacing has been a favored treatment modality for photodamaged and acne-scarred skin over the past several years. Its association with numerous side effects and complications, particularly prolonged erythema and dyspigmentation, however, has dampened the initial enthusiasm reserved for its use. By reducing the laser-associated tissue ablation depth and degree of thermal necrosis, it is possible that the incidence of these side effects can also be reduced. PURPOSE: To evaluate the clinical efficacy and side effect profile of single-pass CO2 laser skin resurfacing in a large series of patients. MATERIALS AND METHODS: A total of 52 consecutive patients (skin photo-types I-VI) with mild facial rhytides,atrophic scars, or infraorbital hyper-pigmentation underwent single-pass treatment with a high energy, pulsed CO2 laser. Side effects to treatment were closely monitored and tabulated. Clinical improvement using a quartile grading scale was assessed independently by two masked medical evaluators at 1, 3, 6, and 12 months postoperatively. RESULTS: Significant clinical improvement was seen in all patients, with peak improvement scores noted 12 months. Greater clinical improvement was seen in patients with darker skin tones despite the near universal incidence of transient postoperative hyperpigmentation in these patients. CONCLUSIONS: Single-pass CO2 laser skin resurfacing can improve the appearance of fine rhytides, mild atrophic scars, and infraorbital hyperpigmentation in all skin types. The severity and duration of side effects and complications are reduced with this technique (compared with multiple-pass procedures)and may offer a possible solution to the problem of treating patients with darker complexions. PMID- 12745599 TI - Erbium:YAG laser skin resurfacing: a Pakistani experience. AB - BACKGROUND AND OBJECTIVES: To evaluate the use of the erbium:YAG laser for resurfacing in type IV skin patients in Pakistan. STUDY DESIGN/MATERIALS AND METHODS: Ten patients with skin type IV underwent laser skin resurfacing with an erbium:YAG laser to treat wrinkles, acne/chickenpox scars and hyperpigmentation. An evaluation was done by the treating physician and a photographic evaluation and grading was done by a blinded observer. RESULTS: The treating physician's records show no erythema, pigmentary alteration, infection or scarring at the 3 month follow-up. The blinded observer's evaluation of the 3-month photographs showed moderate to excellent improvement in 80% of patients in their respective underlying condition. CONCLUSION: The erbium:YAG laser is safe and effective in treating wrinkles, acne/chickenpox scars and hyperpigmentation in skin type IV patients. PMID- 12745600 TI - Treatment of lentigo maligna with combination laser therapy: recurrence at 8 months after initial resolution. AB - This is a case report of an 89-year-old woman with a lentigo maligna treated with alexandrite and ruby lasers. The lentigo maligna never completely resolved despite multiple treatments. Marked cosmetic improvement, how-ever, was obtained. No adverse sequelae have occurred. PMID- 12745601 TI - Delayed response of naevus of Ota to Nd-YAG laser patch test. PMID- 12745610 TI - Therapeutic trials in ALS. PMID- 12745611 TI - Stem therapy for ALS: hope and reality. AB - All are agreed that there is pressing need for an effective treatment for Amyotrophic Lateral Sclerosis (ALS; MND). Such treatment may derive from a combination of therapeutic strategies aimed at different aspects of the disorder, and might include drugs directed at the initial, intermediate or terminal cascade of events leading to cell death, as well as the use of stem cells to replace dead motor neurons, or to protect those that remain. The attraction of cell implantation or transplantation is that it might help to overcome the inability of the CNS to replace lost neurons. It is also clear that neural implantation will yield little benefit if the donor cells fail to integrate functionally into the recipient CNS circuitry. In this respect, ALS poses an especially difficult problem. The recent breakthroughs in stem cell research might nevertheless provide possibilities for neural implantation and cell replacement therapy for patients with ALS. The potential impact of these new approaches to neurodegenerative diseases has been emphasised by the many experiments using human foetal cell grafts in patients affected by Parkinson's and Huntington's disease. Clinical benefits in Parkinson's disease seem to be associated with integration of the donor cells into the recipient brain. Despite promising results, however, significant constraints have hampered the use of foetal cells for neural implantation and transplantation. Besides ethical concerns, the viability, purity, and final destiny of the foetal tissue have not been completely defined. Foetal cells are, in addition, post-mitotic and cannot be expanded or stored for long periods, necessitating close synchronisation of tissue donation and neurosurgery. PMID- 12745612 TI - Can we eliminate placebo in ALS clinical trials? AB - BACKGROUND: Previous studies concluded that the decline in strength in patients with amyotrophic lateral sclerosis (ALS) is a linear function. If so, a patient's natural history might serve as the control, instead of placebo, in a clinical trial. METHODS: A placebo-controlled ALS clinical trial included a natural history phase, followed by a 6-month treatment phase. Each patient's forced vital capacity (FVC) score and maximal voluntary isometric contraction (MVIC) raw scores were measured monthly, standardized, and averaged into megascores. For 138 patients, the arm, leg, FVC, arm+leg combination, and arm+leg+FVC combination megascore slopes during the natural history phase and during the placebo phase were compared. RESULTS: The mean slope of megascores during the natural history phase and the mean slope during the placebo phase were not different for the arm, leg, and arm+leg megascores, but were different for the FVC and arm+leg+FVC combination megascores. CONCLUSIONS: Natural history controls may be useful in ALS exploratory trials that use arm megascore slope as the primary outcome measure. However, there are distinct limitations to the use of natural history controls, so that Phase 3 ALS clinical trials require placebo controls. PMID- 12745613 TI - Reduction of the size of the Golgi apparatus of spinal anterior horn cells in patients with X-linked spinal and bulbar muscular atrophy. AB - The Golgi apparatus (GA) of spinal anterior horn cells was examined immunohistochemically in five patients with X-linked spinal and bulbar muscular atrophy (SBMA), in five patients with sporadic amyotrophic lateral sclerosis (ALS) and in five patients without neurodegenerative diseases. In SBMA cases, reduction of the size of the GA was observed in numerous anterior horn cells; however, fragmentation of the GA, previously described in sporadic and familial ALS with SOD1 mutations, was observed only in a few neurons. In addition, motor neurons bearing an intranuclear inclusion showed a normal network of elements of the GA. The frequencies of fragmented GA in counted motor neurons were 0-2.4 % in SBMA cases, 0-3.0 % in normal control cases and 15.7-55.3 % in ALS cases. The different frequency of fragmented GA between SBMA and ALS adds another finding of pathogenetic difference of neurodegeneration in these two motor neuron diseases. PMID- 12745614 TI - A prospective, randomized, placebo-controlled evaluation of corticoneuronal response to intrathecal BDNF therapy in ALS using magnetic resonance spectroscopy: feasibility and results. AB - During the multicenter, phase III trial of intrathecal BDNF in ALS, we evaluated the neuronal marker N-acetylaspartate (NAA) as a surrogate marker of therapeutic efficacy using proton magnetic resonance spectroscopic imaging (MRSI) in a prospective and blinded manner. Selected subjects tolerated the study well without pump malfunction. The NAA to creatine (Cr) intensity ratio (NAA/Cr) was measured in the precentral and postcentral gyri, the superior parietal lobule, the supplementary motor area, and the premotor cortex. After 4.5+/-0.6 weeks treatment, NAA/Cr did not change significantly in any of the regions in the BDNF treated group (n=5) compared to the placebo group (n=6). The lack of change in NAA correlated with the lack of clinical efficacy and supports the validity of NAA/Cr as a surrogate in this setting. MRSI is a feasible and safe method to evaluate intrathecal therapies in ALS. PMID- 12745615 TI - Health-related locus of control: does it change in motor neurone disease (MND)? AB - OBJECTIVES: Previous studies have attempted to describe locus of control beliefs in people with MND. This exploratory, longitudinal study set out to examine some of the possible correlations of health-related locus of control beliefs and the stability of these beliefs. METHOD: 32 people with Motor Neurone Disease completed the Multi-dimensional Health Locus of Control (MHLC) scale, initially on average 10.3 months after diagnosis, and again on average 16.4 months after diagnosis. Physical symptoms were assessed at both times. RESULTS: Initially there were no correlations between MHLC beliefs or disease duration and physical symptomatology, although longer disease duration was associated with greater beliefs in the role of powerful others in health control. At the second assessment, belief in the role of powerful others controlling health had increased, with this increase relating significantly to a worsening in physical symptoms. At this second assessment, neither duration of symptoms nor time since diagnosis correlated with MHLC beliefs. CONCLUSIONS: Whilst health locus of control beliefs do appear to change in MND, current findings suggest that this does not occur simply as a function of the passage of time. How symptoms change seems to be of particular importance when considering health locus of control beliefs in people with MND. Suggestions are made concerning other factors that might usefully be examined in future studies of this type. PMID- 12745616 TI - Nocturnal pulse oximetry: a new approach to establish the appropriate time for non-invasive ventilation in ALS patients. AB - Previous studies have supported non-invasive ventilation (NIV) in amyotrophic lateral sclerosis (ALS), indicating that it prolongs survival. However, criteria for the use of NIV are yet to be defined. The aim of the present study was to evaluate the potential use of nocturnal pulse oximetry (NPO) as a tool for determining the most appropriate time at which to initiate NIV in ALS patients. We conducted a prospective, comparative, historical-controlled clinical study of 64 consecutive ALS patients. Group 1 (historical control group) comprised 44 patients, 14 women and 30 men, with a mean age of 60+/-13 years, in whom we used NIV after detecting early signs of diurnal respiratory insufficiency (RI). Group 2 had 20 ALS patients, 8 women and 12 men, with a mean age of 56+/-11 years, for whom we started NIV after detecting more than 15 periods of nocturnal desaturation/hour by NPO. All patients were periodically evaluated with Norris spinal and bulbar scores, respiratory function tests (RFT) and NPO at three month intervals for one year. They were subsequently followed until death or loss of autonomy from the ventilator. We compared survival time or time to loss of autonomy from the ventilator between both groups. Survival was longer in Group 2 (p<0.002). We concluded that NPO is a valuable screening test to establish the need for NIV. Our results also suggest that the early diagnosis of RI and the early use of NIV increase patient compliance with NIV. PMID- 12745617 TI - Maximum voluntary isometric contraction: investigation of reliability and learning effect. AB - Maximum Voluntary Isometric Contraction (MVIC) is a standardised, objective and sensitive tool for the measurement of muscle strength. The purpose of this study was to investigate different aspects of reliability of MVIC and to determine if a learning effect existed in a relatively new user of the system. Two clinical investigators participated in the study. The inter- and intra-rater reliability of MVIC of 11 muscle groups was tested on healthy subjects (n=35). Intra-class correlation co-efficients (ICCs) were calculated and the statistical methods described by Bland and Altman were applied to the data. ICCs were higher for the more experienced investigator and a learning effect was demonstrated in a relatively new user of the system. Inter-rater reliability was acceptable but lower than intra-rater reliability. Upper limb tests generally yielded higher ICCs and lower ranges of error. The ICC was similar regardless of whether the maximum or average of the two values was taken in a single session. Utilising the statistical methods proposed by Bland and Altman allows estimation of the magnitude of error of MVIC and gives additional information to the ICC. These methods may be useful in the training of investigators and in clinical interpretation of MVIC values. PMID- 12745618 TI - Amyotrophic lateral sclerosis associated with pregnancy: report of four new cases and review of the literature. AB - Pregnancy in women with ALS is rare and is generally considered a potentially dangerous event. We describe four ALS cases associated with pregnancy, together with a review of the literature. Three of the four women described developed ALS during pregnancy. In three cases a normal delivery was performed, with a healthy child. One patient, with severe respiratory failure, underwent an interruption of pregnancy. Seven other cases are reported in the literature, featuring a total of 11 pregnancies. The association between pregnancy and ALS is quite rare, and a pathogenic relationship cannot be excluded. The pregnancy and the delivery may be normal, but respiratory function should be carefully monitored. Generally, ALS does not have deleterious effects on fetal development. However, pregnancy in a woman with severe respiratory failure may precipitate the disease. PMID- 12745619 TI - Expiratory muscle weakness and assisted cough in ALS. AB - Elimination of airway secretion is a major issue in the care of patients with ALS. Sufficient cough flows have to be generated by expiratory muscles to allow airway clearance. Bulbar and expiratory muscle weakness are often reasons for failure of non-invasive ventilation (NIV) and may lead to tracheostomy. Expiratory aids may help to overcome these problems, at least for some time. We report a patient with advanced ALS, receiving nocturnal NIV, who gained much benefit from regular use of a mechanical in-exsufflation device. PMID- 12745620 TI - Mental retardation associated with Brown-Vialetto-Van Laere syndrome. PMID- 12745622 TI - A simple chaotic neuron model: stochastic behavior of neural networks. AB - We have briefly reviewed the occurrence of the post-synaptic potentials between neurons, the relationship between EEG and neuron dynamics, as well as methods of signal analysis. We propose a simple stochastic model representing electrical activity of neuronal systems. The model is constructed using the Monte Carlo simulation technique. The results yielded EEG-like signals with their phase portraits in three-dimensional space. The Lyapunov exponent was positive, indicating chaotic behavior. The correlation of the EEG-like signals was.92, smaller than those reported by others. It was concluded that this neuron model may provide valuable clues about the dynamic behavior of neural systems. PMID- 12745623 TI - Membrane Na+-K+ ATPase inhibition mediated quantal model for brain evolution. AB - The hypothalamus secretes an endogenous membrane Na+-K+ ATPase inhibitor, digoxin. A digoxin-mediated model of quantal perception is proposed. In the quantal state, self replication of self-organized macromolecules is possible. This leads to the origin of molecular organisms like prions. Macromolecules group together to form organelle, which in evolutionary terms are independent bacteria. The organelle/bacteria symbiotically cluster together to form the cell. The human organism, including the brain, can be visualized as an organized cluster or colony of unicellular, symbiotically grouped flagellated bacteria. Synaptic connections form in the bacterial cluster leading to the evolution of the primitive neuronal networks, and later the human brain. The role of quantal perception and the observer function of consciousness in the origin of matter is important. Symbiotically clustered intergalactic magnetotactic bacterial networks are important in the evolution of the universe. PMID- 12745624 TI - Neuroactive steroids, relaxation, and seizure control. AB - Neuroactive steroids alter the excitability of membrane-bound receptors in the nervous system and have a modulatory role in the stress response and in epileptogenic activity. These changes can be detected in brain as well as in plasma. The resulting rapid (<1 min) action of neuroactive steroids might explain the success of some "alternative" approaches in seizure control. Design requirements for research to adequately examine relaxation training in epileptic patients, as well as corresponding changes in neuroactive steroid levels and seizure frequency, are described. PMID- 12745625 TI - Long term production of reactive oxygen species during perinatal asphyxia in the rat central nervous system: effects of hypothermia. AB - The formation of oxygen-derived free radicals in hypoxic and ischemic/reperfused brains has been proposed as an important step that links brain injury to neuronal death. Previously, we have demonstrated that reactive oxygen species (ROS) production was significantly increased in rat neostriatum during acute perinatal asphyxia (PA) in pups. In this article, we have studied the time course of ROS production in the neostriatum and neocortex of adult rats subjected to PA using electron spin resonance spectrometry (ESR) in order to record ROS production. Further more, we analyzed the actions of hypothermia on ROS release in pups and adult rats. We used for this study 6-month-old rats that suffered sub-severe and severe PA when they were pups. The most significant production of ROS was detected either in the neostriatum or neocortex at 19 and 20 min of PA. Hypothermia during 20 and 100 min at 15 degrees C prevented ROS formation either in pups and adult rats. These data further support the concept that free radicals may contribute to the brain injury alterations and that hypothermia can prevent long-term sequelae induced by PA. PMID- 12745626 TI - Hypothalamic digoxin, hemispheric chemical dominance, and the tridosha theory. AB - Ayurveda, the traditional Indian System of Medicine, deals with the theory of the three tridosha states (both physical and psychological): Vata, Pitta, and Kapha. They are the three major human constitutional types that both depend on psychological and physical characteristics. The Pitta state is described as a critical, discriminative, and rational psychological state of mind, while the Kapha state is described as being dominant for emotional stimuli. The Vata state is an intermediate unstable shifting state. The Pitta types are of average height and built with well developed musculature. The Vata types are thin individuals with low body mass index. The Kapha types are short stocky individuals that tend toward obesity, and who are sedentary. The study assessed the biochemical differences between right hemispheric dominant, bihemispheric dominant, and left hemispheric dominant individuals, and then compared this with the patterns obtained in the Vata, Pitta, and Kapha states. The isoprenoid metabolites (digoxin, dolichol, and ubiquinone), glycoconjugate metabolism, free radical metabolism, and the RBC membrane composition were studied. The hemispheric chemical dominance in various systemic diseases and psychological states was also investigated. The results showed that right hemispheric chemically dominant/Kapha state had elevated digoxin levels, increased free radical production and reduced scavenging, increased tryptophan catabolites and reduced tyrosine catabolites, increased glycoconjugate levels and increased cholesterol: phospholipid ratio of RBC membranes. Left hemispheric chemically dominant/Pitta states had the opposite biochemical patterns. The patterns were normal or intermediate in the bihemispheric chemically dominant/Vata state. This pattern could be correlated with various systemic and neuropsychiatric diseases and personality traits. Right hemispheric chemical dominance/Kapha state represents a hyperdigoxinemic state with membrane sodium-potassium ATPase inhibition. Left hemispheric chemical dominance/Pitta state represents the reverse pattern with hypodigoxinemia and membrane sodium-potassium ATPase stimulation. The Vata state is the intermediate bihemispheric chemical dominant state. Ninety-five percent of the patients/individuals in the tridosha, pathological, and psychological groups were right-handed/left hemispheric dominant, however, their biochemical patterns were different--either left hemispheric chemical dominant or right hemispheric chemical dominant. Hemispheric chemical dominance/tridosha states had no correlation with cerebral dominance detected by handedness/dichotic listening test. PMID- 12745627 TI - Hypothalamic digoxin, cerebral chemical dominance and myalgic encephalomyelitis. AB - The isoprenoid pathway was assessed in 15 patients with chronic fatigue syndrome. The pathway was also assessed in individuals with differing hemispheric dominance to assess whether hemispheric dominance had any correlation with these disease states. The isoprenoid metabolites--digoxin, dolichol, and ubiquinone--RBC membrane Na+-K+ ATPase activity, serum magnesium and tyrosine/tryptophan catabolic patterns were assessed. The free-radical metabolism, glycoconjugate metabolism, and RBC membrane composition was also assessed. Membrane Na+-K+ ATPase activity and serum magnesium levels were decreased while HMG CoA reductase activity and serum digoxin levels were increased in myalgic encephalomyelitis (ME). There were increased levels of tryptophan catabolites--nicotine, strychnine, quinolinic acid, and serotonin--and decreased levels of tyrosine catabolites--dopamine, noradrenaline, and morphine in ME. There was an increase in dolichol levels, carbohydrate residues of glycoproteins, glycolipids, total/individual GAG fractions, and lysosomal enzymes in ME. Reduced levels of ubiquinone, reduced glutathione, and free-radical scavenging enzymes, as well as increased lipid peroxidation products and nitric oxide, were noticed in ME. The biochemical patterns in ME correlated with those obtained in right hemi spheric chemical dominance. The role of hypothalamic digoxin and neurotransmitter induced immune activation, altered glycoconjugate metabolism, and resultant defective viral antigen presentation, NMDA excitotoxicity and cognitive dysfunction, and mitochondrial dysfunction related myalgia in the pathogenesis of ME is stressed. ME occurs in individuals with right hemispheric chemical dominance. PMID- 12745628 TI - Cerebral chemical dominance and neural regulation of cell division, cell proliferation, neoplastic transformation, and genomic function. AB - The study assessed the isoprenoid pathway, digoxin synthesis, and neurotransmitter patterns in individuals of differing hemispheric dominance, neurogenetic disorders, and neoplasms. The HMG CoA reductase activity, serum digoxin, magnesium, tryptophan catabolites, tyrosine catabolites, and RBC membrane Na+-K+ ATPase activity were measured in individuals of differing hemispheric dominance. The digoxin status, membrane Na+-K+ ATPase activity, and serum magnesium were assessed in Huntington's disease, trisomy 21, glioblastoma multiforme, and non-Hodgkin's lymphoma (high grade lymphoma). The results showed that right hemispheric, chemically dominant individuals had elevated digoxin synthesis, increased tryptophan catabolites, and reduced tyrosine catabolites, and membrane Na+-K+ ATPase with hypomagnesemia. Left hemispheric, chemically dominant individuals had the opposite patterns. In neurogenetic disorders and neo plasms also hyperdigoxinemia induced membrane Na+-K+ ATPase inhibition, and hypomagnesemia similar to right hemispheric chemical dominance could be demonstrated. The role of hemispheric chemical dominance and hypothalamic digoxin secretion play a key role in the regulation of cell differentiation/proliferation and genomic function. Ninety-five percent of the patients with neurogenetic disorders and neoplasms were right-handed/left hemispheric dominant by dichotic listening test. However, all of them had biochemical patterns similar to right hemispheric chemical dominance. Hemispheric chemical dominance has no correlation to cerebral dominance detected by handness/dichotic listening test. PMID- 12745629 TI - Hypothalamic digoxin, hemispheric dominance, and neurobiology of love and affection. AB - The human hypothalamus produces an endogenous membrane Na+-K+ ATPase inhibitor, digoxin, which can regulate neuronal transmission. The digoxin status and neurotransmitter patterns were studied in individuals with a predilection to fall in love. It was also studied in individuals with differing hemispheric dominance to find out the role of cerebral dominance in this respect. In individuals with a predilection to fall in love there was decreased digoxin synthesis, increased membrane Na+-K+ ATPase activity, decreased tryptophan catabolites (serotonin, quinolinic acid, and nicotine), and increased tyrosine catabolites (dopamine, noradrenaline, and morphine). This pattern correlated with that obtained in left hemispheric chemical dominance. Hemispheric dominance and hypothalamic digoxin could regulate the predisposition to fall in love. PMID- 12745630 TI - Impact of brain injury severity on personality dysfunction. AB - Although the cognitive effects of traumatic brain injury (TBI) have been well investigated, emotional problems are less well understood. The purpose of this study was to explore the relationship of the length of loss of consciousness (LOC) and the severity of cognitive impairment (HRNB) to personality changes after head trauma. The subjects included 320 chronic TBI patients. A multivariate analysis was conducted using HRNB (three levels), as measured by a modified Halstead-Reitan Impairment Index and LOC (four levels), as the independent variables, and 30 scales from the Minnesota Multiphasic Personality Inventory-2 (MMPI-2) as the dependent variables. Multivariate Fs were significant for both HRNB and LOC. Follow-up ANOVAs and t-tests revealed a consistent pattern of findings across both variables. The data indicated that the reported emotional problems increased with the severity of both LOC and HRNB, except in the most severe groups. The severe subjects when defined by either variable displayed high levels of denial and a lack of awareness of their problems. LOC and HRNB contributed independently to the degree of personality problems, however, LOC was found to be a stronger predictor of personality change than was HRNB at milder levels of severity. The implications of these findings for both treatment and understanding of the mechanisms of head injury are discussed. PMID- 12745631 TI - Anxiety and trail making test scores in a sample of cocaine abusers. AB - Anxiety effects on the Trail Making test (TMT), a test often used for screening for cognitive impairments, were examined in a sample of cocaine abusers in drug abuse treatment programs. A mixed race sample of 4306 subjects was drawn from electronic files of data from the Drug Abuse Treatment Outcome Study (DATOS). The DATOS was a naturalistic, prospective cohort study that collected data from 1991 1993 in 96 programs in 11 cities in the United States. Data were analyzed to determine the effects of anxiety on the TMT scores A and B, and also derived indices created by adding, subtracting, multiplying, and dividing parts A and B of the TMT in this large treatment sample of cocaine abusers. The variables of sex, age, ethnicity, and education were included in analyses to control for demographic effects. The ratio derived score was the least sensitive TMT score to the effects of anxiety, but all TMT R-squares were quite small. PMID- 12745632 TI - Targeted expression of temperature-sensitive dynamin to study neural mechanisms of complex behavior in Drosophila. PMID- 12745633 TI - Doublesex gene expression in the central nervous system of Drosophila melanogaster. AB - Despite several behavior-genetic studies that have suggested roles played by doublesex (dsx) in neural tissues, it has not been demonstrated that the products of this gene are actually present in the central nervous system (CNS). In this report, we describe the cellular, spatial, and temporal expression patterns of dsx gene products in the developing and adult CNS by applying RT-PCR and immunohistochemical procedures. dsx gene products were detected in the CNS of 3rd instar larvae, pupae, and adults. DSX-immunoreactive signals were observed within the brain and in both the thoracic plus abdominal ganglia of the ventral nerve cord. Most, but not all, cells inferred to contain DSX proteins (by the results of genetic controls for antibody specificity) were further determined to be neurons (by coexpression of a protein that marks such CNS cell types). Temporally varying expression of DSX was most prominently observed in the rapidly metamorphosing early and mid-pupal stages, suggesting that this gene contributes to establishment of sexually dimorphic neuronal structures which subserve adult sexual behaviors. Elements of the spatial and temporal patterns of DSX immunoreactivity also imply that sexually dimorphic dsx expression in certain neuronal clusters within the adult CNS could participate in ongoing operations of the mature nervous system with respect to the courtship behaviors that are affected by dsx mutations. PMID- 12745634 TI - Altered drug resistance and recovery from paralysis in Drosophila melanogaster with a deficient histamine-gated chloride channel. AB - The recent identification and characterization of two genes, encoding histamine gated chloride channel subunits from Drosophila melanogaster, has confirmed that histamine is a major neurotransmitter in the fruitfly. One of the cloned genes, hclA (synonyms: HisCl-alpha1; HisCl2), corresponds to ort (ora transientless), mutationsin which affect synaptic transmission in the Drosophila visual system. We identified a mutational change (a null mutation) in the genomic and RNA copies of hclA derived from mutants carrying the ort(1) allele. This correlates with new phenotypes observed in the mutant strain. We found hypersensitivity to the avermectin neurotoxins in both the ort(1) adult flies and third instar larvae compared to Oregon R wild-type animals. On the other hand, the mutation makes both male and female adult flies more resistant to treatment with diethyl ether, and the animals show substantially prolonged recovery from paralysis after diethylether anaesthesia, as well as from paralysis after mechanical shock, as revealed by the bang sensitivity test. Altogether, our data give direct evidence that in vivo a HCLA subunit-containing receptor has a distinct role in the neurotoxic action of the avermectins. They also provide new evidence for a function in the response to diethylether anaesthesia and, moreover, that HCLA function is not limited to the visual system. PMID- 12745635 TI - mRNA expression of the lipid and mechano-gated 2P domain K+ channels during rat brain development. AB - mRNAs encoded by genes for the lipid-sensitive mechano-gated K(+) channels TREK 1, TREK-2, and TRAAK were detected in rat brain at different life-cycle stages: 18-day embryos, postnatal days 1, 7, 28, and 60 (adulthood). mRNA expression of TREK-1 or TREK-2 showed no appreciable changes during the development of cortex and hippocampus. TRAAK mRNA expression increased with development and reached an apparent maximum at postnatal day 28 in hippocampus and day 60 in cortex. These data suggest that TRAAK might be important in the development of rat brain. PMID- 12745636 TI - Cash income, intrahousehold cooperative conflict, and child health in central Mozambique. AB - This study presents qualitative data on individual cash income generation and intrahousehold bargaining in a sample of 100 households in central Mozambique. It is now recognized that intrahousehold resource allocation patterns can be critical determinants of children's health in the developing world. Recently developed "bargaining-power" models suggest that individual incomes are often not pooled in households and that decisions are the result of a bargaining process that involves cooperation and conflict between men and women. Women's income, many believe, is more often spent on child welfare. Development projects should target benefits to women for greater impact on child health. Some argue that households consist of separate, gendered spheres of economic responsibility that intersect through a "conjugal contract" that defines the terms of cooperation. The findings here support the "separate-spheres" depiction of the household and reveal women's subordinated position in the external cash economy, which undermines their intrahousehold bargaining power. PMID- 12745637 TI - Sweet blood and social suffering: rethinking cause-effect relationships in diabetes, distress, and duress. AB - I draw upon anthropological engagements with bioscience and embodiment in order to unpack current approaches to defining and preventing diabetes mellitus. The analysis stems from the conviction that carefully considering the symbolic frames through which we conceive of diseases, their origins, their distribution, and their consequences will assist us in planning and implementing interventions to improve population health. I argue that research and interventions focused on the sweetness of blood would benefit from rethinking intersections between diabetes, distress, and duress. In many instances, the lived experience of diabetes is consonant with an understanding of distress (i.e., "social suffering") that expands conventional understandings of population health problems. Diabetes incidence is rising worldwide, but it is rising especially rapidly in Aboriginal and other disadvantaged populations. Notably, diabetes is now three to five times more common in Canada's First Nations population than it is in its non-Aboriginal population. Yet as recently as 50 years ago, diabetes and associated health problems were rare in these groups. To come to grips with such transformations and disparities is to advance the population health research agenda. PMID- 12745638 TI - Gender schema and prostate cancer: veterans' cultural model of masculinity. AB - Coming to terms with disease, chronic illness, and aging may be challenging for men who adhere to an inflexible gender schema. In this study of elder U.S. veterans' ideas about masculinity, we find that prostate cancer patients reaffirm a strongly moral normalizing discourse about "being a man" yet tend to separate roles and values from male physical and sexual attributes. Using systematic data collection methods taken from cognitive anthropology, we map veterans' schema of masculinity and examine the relative importance that cancer patients and non patients give to gender attributes. The results demonstrate the complementarity between cognitive and narrative approaches in medical anthropology. This research also suggests the hypotheses that (1) coming to terms with iatrogenesis may involve a subtle reformulation of masculinity and that (2) men with a fixed view of masculinity may have worse health outcomes than do those who accept the changes accompanying their treatment for prostate cancer. PMID- 12745640 TI - Chemokines and antitumor immunity: walking the tightrope. AB - Chemokines play an important role in the generation of the immune system and in virtually every aspect of an immune response. The role of chemokines in antitumor immunity has been less straightforward to discern. A dichotomy exists in the field. One area of research has focused on the impact of tumor-derived chemokines, implicating them in everything from metastases to immune suppression. Another area of research has been dedicated to the introduction of chemokines into tumor cells in order to facilitate immune cell recruitment. In this review these two areas of investigation will be explored. PMID- 12745641 TI - Redirecting T lymphocyte specificity using T cell receptor genes. AB - Redirecting T cells by transferring T cell receptor (TCR) genes from tumor associated antigen (TAA)-reactive T cell clones into human peripheral blood lymphocytes (PBL) has therapeutic potential for the treatment of diseases, including cancer. T cell specificity can be altered using retroviruses encoding TCRalpha and TCRbeta chain genes, or chimeric immunoglobulin (cIg) genes containing signaling domains of CD3 zeta or Fc epsilon RI-gamma. This review evaluates recent studies using TCRs and cIgs to redirect T cell specificity and discusses some of the technical and biological hurdles that need to be addressed before these approaches can be successfully used to treat patients. PMID- 12745642 TI - Current status of dendritic cell immunotherapy of malignancies. AB - Because dendritic cells (DC) are central to the induction of antigen-specific T cell responses, their use for the active immunotherapy of malignancies has been of considerable interest. Since clinical trials with DC-based vaccines have been initiated, a number of important developmental issues have become apparent. These include the ideal source and type of DC, the form of antigen and method of loading DC, whether to induce maturation, the route and timing of immunization, and the optimal clinical scenario. Clinical responses such as stability of disease and tumor regressions have been reported in some patients, particularly with melanoma, myeloma, and prostate cancer. PMID- 12745643 TI - Monitoring immune responses in cancer patients receiving tumor vaccines. AB - Clinical evaluation of therapeutic tumor vaccines has resulted in examination and comparison of the types of immune function assays required to monitor tumor antigen-stimulated T cell effector function in immunized patients. Three of the most commonly used assays include ELISPOT, tetramer assay, and cytokine flow cytometry (CFC). Discussed are the method and principles for each assay and an assessment of important methodological, reagent, and data acquisition issues that are relevant for the accurate and effective use of the assays. The sensitivity and utility of the assays and present arguments advocating their integrated use in future immunomonitoring studies are also discussed. PMID- 12745644 TI - Spontaneous regression of Hodgkin's disease: two case reports and a review of the literature. AB - We present two cases of patients with Hodgkin's lymphoma who experienced spontaneous regressions of their disease. The first case was a 31-year-old man diagnosed with stage IIIA lymphocyte predominant Hodgkin's disease in 1994, who elected to be followed without any treatment. Over the subsequent 3 years, he experienced significant regression in his lymphadenopathy, and still remains asymptomatic of his disease 70 months after diagnosis. The second case was a 47 year-old man with a bulky anterior mediastinal mass found on a thoracic CT scan, ultimately diagnosed with stage IIB Nodular Sclerosing Hodgkin's Lymphoma. Repeat imaging of the chest performed two months later, just prior to initiating treatment, revealed that the mass had spontaneously decreased by >75% of its original size. Spontaneous regressions of Hodgkin's lymphoma are exceedingly rare. A review of the literature regarding spontaneous regressions of lymphoma and cancer in general is discussed. PMID- 12745645 TI - Cancer therapy: new targets for chemotherapy. AB - The number two cause of mortality in developed countries is cancer. Despite the enormous effort put into cancer prevention, early diagnosis and treatment, it is likely that the incidence of the cancer morbidity and mortality will increase for the foreseeable future. This is due to various factors such as increased life expectancy, changes in environment and also the socio-economic situation around the world. Some cancer attracts more attention than others and increasingly epidemiological information is reaching the general public and is beginning to influence behavior. It is now well recognized that, for example, 1 of 8 women in the industrialized world will be diagnosed with breast cancer. Additionally, a strong correlation was established between lung cancer incidence and smoking and it is broadly accepted that the incidence of colon cancer is directly related to age and diet, and has been increasing over time. The current failure of preventive measures to significantly reduce the increasing incidence of these common tumors illustrates the importance of effective cancer treatment strategies, including chemotherapy. The combination of various anticancer drugs, given together with surgery and radiotherapy, gives hope to many patients. There has been recent evidence of improved therapeutic outcome with recent approaches and newer agents but for continuing effective chemotherapeutic treatment there is a need for a detailed understanding of their mechanisms of action and on the rationale of their application. This review attempts to provide up-to-date information regarding the development of new and innovative treatment strategies for cancer chemotherapy. Virtually, every year several of new targets for cancer therapy on both, cellular and molecular levels, are identified and new drugs enter not only clinical trials but also are included in well accepted and documented therapeutic protocols. As this review is in addition to our review published previously (Medical Principles and Practice 11, 2002, 117-125), we have tried to include new and innovative targets and drugs that attract attention at present. Although it is not possible to provide a complete list of all achievements and cover all work done in this field, we hope to be able to give some insight into this rapidly developing area. PMID- 12745646 TI - Imatinib Mesylate (Gleevec) is a useful agent in the salvage treatment of adults with relapsed/refractory Philadelphia positive acute leukemias. AB - Philadelphia chromosome-positive (Ph+) acute leukemias have a markedly poor prognosis when treated with conventional chemotherapy alone. Even with intensive treatment such as allogeneic transplant, a large proportion of patients relapse. We describe here four cases of relapsed/refractory Ph+ acute leukemias who were treated with Imatinib Mesylate (Gleevec) as monotherapy. Significant clinical and molecular responses were observed in these patients, which allowed us to deliver highly intensive treatments such as second allogeneic stem cell transplant and matched unrelated transplant in these patients. Gleevec may prove to be a useful agent in the salvage therapy of such patients. PMID- 12745647 TI - Outcome and toxicity of salvage treatment on patients relapsing after autologous hematopoietic stem cell transplantation--experience from a single center. AB - Patients with hematological malignancies who relapse after autologous stem cell transplantation (auto-SCT) generally have poor prognosis. Salvage treatment is often associated with severe toxicities. The aim of our study was to evaluate retrospectively the toxicity and outcome of rescue therapy in patients with acute leukemias, non-Hodgkin's lymphoma (NHL), Hodgkin's disease (HD) and multiple myeloma (MM) relapsing after auto-SCT. Fifty-four of the 62 patients who relapsed received some form of salvage chemotherapy. Six (10%) patients were treated by second stem cell transplantation, which was allogeneic in 5 cases. Toxicity of the salvage therapy was significant. As a result of adverse effects, salvage therapy had to be discontinued or reduced in 14 patients (26%). The outcome of salvage was evaluated after 90 days. Of the treated patients, 14 (26%) entered into complete remission with another 5 (9%) reaching partial response. The disease was stabilized in 5 patients (9%) but 30 (56%) patients were in progression or dead. Overall survival of the patients was poor with the median survival of 8.7 months after relapse and the leading cause of death being progressive disease. In conclusion, the development of new, more efficient regimens is critical if disease-free survival is to be increased in patients who relapse after auto SCT. PMID- 12745648 TI - Decreased transfusion requirements in patients given stem cell allografts using a non-myeloablative conditioning regimen: a single institution experience. AB - We report our experience of allogeneic peripheral blood stem cell transplantation using non-myeloablative conditioning regimens delivered and supported on an outpatient basis. A group of 44 patients underwent 47 allograft procedures using peripheral blood stem cells. Approximately one third of the individuals did not require red blood cells transfusions: the median of transfused red blood cells units was 1 (range 0-10). In addition one out of three did not require platelet transfusions either, the median of platelet transfusions being 1 (range 0-6). In fourteen allografts (30%) neither red blood cells nor platelet transfusions were used. An inverse correlation was found between the number of CD34 cells infused and the PRBC and PLT transfusion requirements, those patients receiving high numbers of CD34 cells needing fewer transfusions of both PRBC and platelets. The possibility of conducting allografts without transfusion of blood products in some patients may result in a decrease in both cost and the risks stemming from exposure to human blood derivatives. PMID- 12745650 TI - Neutrophil elastase mutations in congenital neutropenia. AB - Severe congenital neutropenia (SCN) was originally described as an autosomal recessive disorder. Autosomal dominant and sporadic forms of the disease have subsequently been recognized. All forms of the disease are manifest by persistent severe neutropenia and recurrent bacterial infection. Cyclical neutropenia (CyN) is characterized by periodic neutropenia inter-spaced with (near) normal neutrophil counts. Recently, heterozygous mutations in the ELA2 gene encoding neutrophil elastase (NE) have been described in the majority of cases of CyN and sporadic and autosomal dominant SCN. A case of paternal mosaicism has provided genetic "proof" of the pathogenicity of such mutations, but the exact pathogenic mechanism remains elusive. This review will focus on the mosaic proof and examine possible pathogenic mechanisms. The lack of obvious associations and indeed overlap between the mutations that cause the two diseases will also be discussed. Clinically to date, the discovery of an elastase mutation has been of limited value to individual patients. However, it is hoped that further genotype/phenotype studies may improve assessment of patient prognosis. PMID- 12745649 TI - Kinetics of hematopoiesis in bone marrow cultures from patients with chronic myeloid leukemia: effect of recombinant cytokines in dexter-type long-term cultures. AB - Chronic myeloid leukemia (CML) is a hematological neoplasia that results from the transformation of a hematopoietic stem cell. It is characterized by the expansion of the myeloid lineage, which results in the accumulation of mature and immature granulocytes in peripheral blood and bone marrow. However, when CML marrow cells are cultured in Dexter-type long-term cultures (LTMC) hematopoiesis is defective and can be sustained for only a few weeks. One possible explanation for the deficient growth of hematopoietic cells in CML LTMC is that some factors that act as key regulators of hematopoiesis are absent in this experimental system. Thus, we tested this hypothesis by adding recombinant cytokines to these cultures. As a first approach, we added recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF), rhGranulocyte-CSF (rhG-CSF) and rhErythropoietin (rhEPO); each factor was added individually once a week. Addition of rhGM-CSF and rhG-CSF resulted in a significant increase in the levels of nucleated cells and myeloid progenitors; the highest effects were seen in the presence of rhGM-CSF. Interestingly, such a cytokine also induced a significant decrease in the levels of erythroid progenitors. Recombinant hEPO had no significant effects on nucleated cells or myeloid progenitors, however, it induced a significant, although transient, increase in the levels of erythroid cells. The above results indicate that the hematopoietic regulators used here (rhGM-CSF, rhG-CSF and rhEPO) are capable of stimulating the growth of hematopoietic cells in LTMC from CML patients. Thus, this study demonstrates that it is, indeed, possible to manipulate CML LTMC by the addition of recombinant cytokines; this observation may be of particular relevance, since this in vitro experimental system has already been used as a method for purging of leukemic cells in autologous transplant settings. By using specific recombinant hematopoietic modulators it might be possible to make LTMC a more efficient system for such a clinical purpose. PMID- 12745651 TI - Application of the molecular analysis of the T-cell receptor repertoire in the study of immune-mediated hematologic diseases. AB - The basis for the vast recognition spectrum of the T-cell receptor (TCR) can be determined by the rearrangement and recombination of the variable, diversity and joining regions of the variable portions of beta (B) and alpha (A) chains as well as their recombination and modification. Analysis of the TCR rearrangement has been routinely used to detect clonality for the diagnosis of lymphoid malignancies. However, molecular analysis of the TCR repertoire can be a powerful tool in the study of T-cell responses to pathogens and in autoimmune diseases. The concept of the oligoclonality in the context of cellular immune responses is based on the presence of immunodominant T-cell clones within distinct T-cell subpopulations used for analysis. Under normal circumstances, a limited number of clones undergo periodic expansions in reaction to foreign antigens. Under pathologic conditions, though, the derailment of immune regulation allows expansions of specific and potentially pathogenic T-cell clones. For example, large granular lymphocyte (LGL) leukemia illustrates an extreme expansion of a single T-cell clone associated with a distinct autoimmune pathology, which suggests an exaggerated clonal response to a specific antigenic target. In immune mediated bone marrow failure syndromes, clonal rearrangement of the TCR cannot be detected in unseparated blood or marrow. Nevertheless, individual T-cell clones can significantly expand and may allow for demonstration of oligoclonality in selected T-cell populations. These subpopulations are defined, for example, by a specific beta (B)-chain usage or other phenotypic markers. Given the diversity of the TCR recognition spectrum, the task of identifying immunodominant clonotypes derived from unique complementarity determining region-3 (CDR3) sequences is very complex. However, expanded T-cell clones likely represent immunodominant responses which can be detected on the molecular level using analysis of the individual TCR VB-chain representation, CDR3 size fragment skewing, and determination of the frequency of individual clonotypic sequences. In the future, TCR VB clonotypes may be applied as a diagnostic tool, analogous to serologic markers. As an investigative tool in hematology, molecular analysis of the TCR utilization pattern and the detection of immunodominant clonotypes represents a novel approach in the study of immune-mediated hematologic diseases, such as aplastic anemia (AA), some forms of myelodysplasia (MDS), anti-leukemic immune surveillance, graft-versus-leukemia effects and graft-versus-host disease (GvHD). PMID- 12745654 TI - Current clinical practice. DIC 2002: a review of disseminated intravascular coagulation. AB - The turn of the millennium has seen clear advances in the understanding and management of Disseminated Intravascular Coagulation (DIC). The recognition that its pathogenesis stems from sustained thrombin generation in fuelling the cycle between inflammation and coagulation has seen the first successful treatment in severe sepsis through targeting this activity. An advance in treatment brings heightened relevance to laboratory testing, which now emphasises earlier detection and better monitoring to facilitate improved risk-identification and assessment of therapeutic efficacy. This review article also provides insights into future strategies that might build on the foundation of improving prognosis for the patient with DIC. PMID- 12745652 TI - c-Myc oncogene and Cdc25A cell activating phosphatase expression in non-Hodgkin's lymphoma. AB - The product of proto-oncogene c-Myc is a potent activator of cell proliferation. The prognostic importance of the over expression of c-Myc and its transcriptional target Cdc25A in non-Hodgkin lymphoma (NHL) patients remains to be elucidated. To determine the role and the prognostic relevance of c-Myc and Cdc25A over expression in this group, we analyzed the expression of c-Myc oncoprotein by immunohistochemistry and Cdc25A mRNA by reverse-transcription polymerase chain reaction (RT-PCR) in the biopsied lymph nodes of 59 NHL patients. Over expression of c-Myc oncoprotein (P62) was observed in 32 out of 59 samples (54.2%) and Cdc25A in 36 out of 59 (60.1%). The percentage of c-Myc oncoprotein and Cdc25A mRNA over expression was significantly increased from low grade (4/12=25%, 4/16=25%) through intermediate grade (9/20=45%, 10/20=50%) to high grade lymphoma (19/23=82.6%, 22/23=95.6%) respectively (P=0.001 for both). The proportion of patients with positive c-Myc and Cdc25A over expression was significantly higher among patients with elevated serum lactic dehydrogenase (sLDH), and serum beta 2 microglobulin compared to those with normal levels (P<0.05, <0.01, respectively). Moreover, 80 and 90% of NHL patients with bone marrow infiltration at diagnosis had c-Myc and Cdc25A over expression, respectively. On the other hand, positive c Myc, and Cdc25A over expression were not significantly related to the grade of international prognostic index, or the presence of B symptoms or to histopathological type. The expression of c-Myc and Cdc25A was significantly elevated in those who died when compared to survivors (P<0.001 for both). Moreover, positive c-Myc and Cdc25A over expression was associated with shortened overall survival. IN CONCLUSION: over expression of c-Myc and Cdc25A may be poor prognostic factor in NHL and associated with poor outcome. Assessments of c-Myc and Cdc25A expression in NHL at diagnosis are likely to be helpful in predicting patient outcome and selecting optimal therapeutic regimen. PMID- 12745655 TI - Factor V Leiden detection by polymerase chain reaction-restriction fragment length polymorphism with mutagenic primers in a multiplex reaction with Pro G20210A--a novel technique. AB - Factor V Leiden (FVL) R506Q and Prothrombin G20210A are clinically important genetic mutations associated with increased susceptibility to venous thrombosis. The objective of our study was to design a polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP) reaction that allows simultaneous detection of these two mutations. The reaction can be used in routine diagnostic settings. We have analysed 4504 alleles for each mutation with a mutagenic primer based PCR system with a low failure rate. The system eliminates the false positive FVL G1691A results associated with other PCR/RFLP caused by rare confounding mutations adjacent to restriction endonuclease recognition sites. This multiplex PCR/RFLP reaction is rapid, robust and dependable. PMID- 12745656 TI - New strategies for BMT, organ transplantation, and regeneration therapy. AB - Bone marrow transplantation (BMT) is becoming a powerful strategy for the treatment of hematologic disorders, congenital immunodeficiencies, metabolic disorders and also autoimmune diseases. We have previously found using various animal models for spontaneous autoimmune diseases, that allogeneic bone marrow transplantation (allo BMT) can be used to prevent and treat various autoimmune diseases. In addition, we have found that autoimmune diseases are stem cell disorders. However, in MRL/lpr mice, which are radiosensitive (<8.5 Gy), we found that conventional BMT had only a transient effect on autoimmune diseases, which were found to recur. Therefore, we concentrated on discovering new strategies to prevent and treat autoimmune diseases in the radiosensitive and chimeric resistant MRL/lpr mouse. Using MRL/lpr mice, we established a new method for allo BMT. In this method, whole bone marrow cells (BMCs), containing a small number of T cells and mesenchymal stem cells (MSCs), were directly injected into the bone marrow cavity (intra-bone marrow [IBM]-BMT). MRL/lpr mice treated with IBM-BMT survived more than 2 years without showing the symptoms of autoimmune diseases. To apply this BMT method to humans, we have also established a new method for BMC harvesting using cynomolgus monkeys. In this method, BMCs are harvested from the long bones using a "Perfusion Method" (PM) and the whole BMCs (including MSCs) are then injected directly into the IBM. We believe that this new method will become a powerful strategy for the treatment of various intractable diseases, including age-associated diseases such as osteoporosis. PMID- 12745658 TI - Role of plasminogen activator inhibitor-1 (PAI-1) levels in the diagnosis of BMT associated hepatic veno-occlusive disease and monitoring of subsequent therapy with defibrotide (DF). AB - Hepatic veno-occlusive disease (VOD) is a common and potentially fatal complication of high dose chemotherapy with allogeneic/autologous stem cell transplant (SCT). The diagnosis and treatment of hepatic VOD is controversial. Clinical features are non-specific and may be mimicked by a number of other conditions causing hyperbilirubinaemia post-transplantation. Plasminogen activator inhibitor-1 (PAI-1) has been proposed as a specific marker of VOD [1]. Defibrotide (DF) is a polydeoxyribonucleotide, which has been found to have anti thrombotic, anti-ischaemic and thrombolytic properties without causing significant anti-coagulation. Recent evidence [2,3] suggests that use of DF in patients with severe VOD results in a promising response rate without attributable significant toxicity. Between January 1998 and July 1999, PAI-1 levels were measured serially in 16 patients undergoing SCT who had subsequently developed hyperbilirubinaemia. Diagnosis of VOD was made by established clinical criteria [4,5]. At the time of diagnosis, PAI-1 levels (mean+/-SD) were significantly elevated in patients with VOD (90.7+/-47 ng/ml, n=7) when compared with patients with jaundice from other causes post transplantation (12.1+/-6.4 ng/ml, n=9). Five of the patients with VOD received treatment with DF. Four out of five patients showed an initial response to DF (significant fall in bilirubin and improvement in other signs/symptoms) with one of these patients having a complete response (bilirubin < 2.0 mg/dl and full resolution of signs/symptoms and end-organ toxicity). Following treatment with DF, a corresponding fall in PAI 1 levels was noted in those responding, with non-responders maintaining raised levels. CONCLUSION: Raised PAI-1 levels post stem cell transplant are specific for VOD and a subsequent decrease in levels following treatment with DF may be associated with response to treatment. PMID- 12745657 TI - Autologous peripheral blood stem cell transplantation in first remission adult acute myeloid leukaemia--an intention to treat analysis and comparison of outcome using a predictive model based on the MRC AML10 cohort. AB - The role of autologous peripheral blood stem cell transplantation (APBSCT) in acute myeloid leukaemia (AML) remains controversial. The current study evaluated the application of APBSCT in a large consecutive series of patients with untreated AML, and compared outcome with a predictive model based on MRC AML10 data. Of 148 evaluable patients, 118 patients entered complete remission (CR) after induction therapy comprising three cycles of daunorubicin, cytosine arabinoside and oral 6-thioguanine. Of these patients, 68 (57%) proceeded to consolidation therapy with two courses of intermediate dose cytosine arabinoside, and stem cell mobilisation, and 40 of these patients (34%) underwent the APBSCT procedure after high dose busulphan conditioning. Harvest quality was the main factor precluding APBSCT. Five-year event-free survival (EFS) in patients who achieved CR was 38% and in APBSCT patients was 57%. There were no transplant related deaths. No significant differences were demonstrated between observed and expected outcomes at 1 and 2 years, based on the predictive model derived from the MRC AML10 study. These data therefore indicate that only a third of eligible adult patients will undergo APBSCT. However, the results demonstrate favourable survival in such patients, with no transplant-related mortality. PMID- 12745659 TI - Dendritic cell-based immunotherapy for the treatment of hematological malignancies. AB - Dendritic cells (DCs) are professional antigen-presenting cells and are frequently used in current immunotherapy protocols. The administration of DCs loaded with tumor-associated proteins or peptides results in the induction of immune responses against different types of malignant cells. Methods for large scale generation of DCs in a sufficient quality and quantity have permitted their use in clinical experiments. DC-based vaccines have already shown promise in follicular non-Hodgkin's lymphoma, and to some extent, in other hematological malignancies. Several strategies have been developed to boost their potency as a new and relatively non-toxic treatment modality. Our review focuses on clinical trials using DCs in the treatment of hematologic malignancies and on recent studies of the immunophenotype, development, and maturation of DCs may have an important impact on designing DC-based antitumor vaccines. PMID- 12745660 TI - Acute promyelocytic leukemia: a case-based review. AB - Retinoid therapy for acute promyelocytic leukemia (APL) is one of the major achievements of leukemia research in the last 15 years. Use of all trans retinoic acid (ATRA) has changed the prognosis of APL from a fatal leukemia to a highly curable disease. This case-based review examines the available clinical and scientific data to form evidence-based decisions in the management of APL. The main aim of this review is to highlight recent progress made in the management of APL and address the role of maintenance therapy, prognostic factors for relapse and treatment of relapsed disease. PMID- 12745661 TI - Type J CBFbeta/MYH11 transcript in the M4Eo subtype of acute myeloid leukemia. AB - Acute myeloid leukemia (AML) carrying inversion or translocation of chromosome 16 is usually associated with the FAB M4Eo morphological subtype and belongs to AMLs with a relatively favorable prognosis. At the molecular level, it is associated with a disease-specific fusion gene, CBFbeta/MYH11. Previously, 10 different types of CBFbeta/MYH11 fusion transcripts have been described in the literature, 7 of them are still known as unique cases. In the current study, peripheral blood and/or bone marrow samples from 265 AML patients were tested for the presence of the CBFbeta/MYH11 fusion using RT-PCR and 12 (4.5%) positive cases were identified. The most common type A CBFbeta/MYH11 transcript was confirmed in 11 patients. The transcript in the remaining one (a 71-year-old female) was different and sequence analysis allowed us to classify it as CBFbeta/MYH11 type J. In contrast to the first type J case previously reported from Australia, this patient exhibited a typical FAB M4Eo morphology. The evidence of the second case indicates that the type J breakage might be a non-random event within the MYH11 gene. PMID- 12745663 TI - Hematology morphology forum case 2. Mycobacterial spindle-cell pseudotumor of the lymph nodes. PMID- 12745662 TI - Eradication of invasive mucormycosis--effectiveness of the Echinocandin FK463. AB - BACKGROUND: Invasive rhinocerebral mucormycosis is a rare and often fatal opportunistic fungal infection. It is encountered in immunocompromised hosts exemplified by those with diabetes, human immunodeficiency viruses and particularly haematologic malignancies typically after high-dose chemotherapy and stem cell transplantation. In contrast to the more usual outcome with rapid progression and death. We now describe a successful eradication attributable to the use of a newly available antifungal agent. SETTING: Haematology department and bone marrow transplantation unit. MATERIAL AND METHOD: Two patients are contrasted. The first with acute leukaemia developed rapidly progressive facial swelling with mucormycosis proven on biopsy. Treatment over 2 months with maximally tolerated doses of amphotericin failed to halt intracranial extension and death resulted. The second, presented with acute lymphoblastic leukaemia in August 1997, underwent successful autologous bone marrow transplantation in February 1998. Relapse followed in March 1999 and after reinduction and consolidation receive a matched unrelated volunteer allograft in September 1999. A second recurrence was documented in April 2000 and in spite of achieving remission he developed a fever that was managed empirically with intravenous amphotericin and, on discharge, oral itraconazole. Left-sided facial swelling expanded rapidly and biopsy showed extensive invasion of the maxillary sinus with mucormycosis. FK463 was added on 5 June 2000 with gradual reduction in facial pain and within 1 month all clinical signs and resolved. Serial biopsies that included histopathologic investigation and microbiologic cultures confirmed eradication of the invasive mucor. In view of the potential danger of recrudescence this treatment regimen was continued through further chemotherapy and, once again disease-free, a second matched unrelated volunteer allograft took place in August 2000. Full reassessment at the time failed to demonstration any residual fungus. Engraftment was confirmed but neutropenic sepsis resulted in severe inflammatory response syndrome with progression to multiple organ dysfunction to which he succumbed without any evidence of leukaemic or systemic mycosis. CONCLUSION: Echinocandin FK463 is of documented value in managing invasive candidiasis and aspergillosis. This is believed to be the first case of successful outcome with one of the angiotrophic zygomycetes. PMID- 12745664 TI - Beta-cell expansion for therapeutic compensation of insulin resistance in type 2 diabetes. AB - Insulin resistance is the primary cause of type 2 diabetes. However, if compensated by increased insulin production, insulin resistance by itself does not lead to overt disease. Type 2 diabetes develops when this compensation is insufficient, due to defects in beta-cell function and in regulation of the beta cell mass. beta-Cell transplantation, as well as approaches that replenish or preserve the endogenous beta-cell mass, may facilitate the treatment of type 2 diabetes in patients requiring exogenous insulin. PMID- 12745665 TI - Ins1 gene up-regulated in a beta-cell line derived from Ins2 knockout mice. AB - The authors have derived a new beta-cell line (betaIns2(-/-lacZ)) from Ins2-/- mice that carry the lacZ reporter gene under control of the Ins2 promoter. betaIns2(-/-lacZ) cells stained positively using anti-insulin antibody, expressed beta-cell-specific genes encoding the transcription factor PDX-1, glucokinase, and Glut-2, retained glucose-responsiveness for insulin secretion, and expressed the lacZ gene. Analysis of Ins1 expression by reverse transcriptase-polymerase chain reaction (RT-PCR) showed that Ins1 transcripts were significantly raised to compensate for the lack of Ins2 transcripts in betaIns2(-/-lacZ) cells, as compared to those found in betaTC1 cells expressing both Ins1/Ins2. Thus, transcriptional up-regulation of the remaining functional insulin gene in Ins2-/- mice could potentially contribute to the beta-cell adaptation exhibited by these mutants, in addition to the increase in beta-cell mass that we previously reported. We have also shown that lacZ expression, as analyzed by determining beta-galactosidase activity, was up-regulated by incubating betaIns2(-/-lacZ) cells with GLP-1 and/or IBMX, 2 known stimulators of insulin gene expression. These cells thus represent a new tool for testing of molecules capable of stimulating Ins2 promoter activity. PMID- 12745666 TI - The effects of middle cerebral artery occlusion on central nervous system apoptotic events in normal and diabetic rats. AB - Apoptosis and neural degeneration are characteristics of cerebral ischemia and brain damage. Diabetes is associated with worsening of brain damage following ischemic events. In this study, the authors characterize the influence of focal cerebral ischemia, induced by middle cerebral artery occlusion, on 2 indexes of apoptosis, TUNEL (terminal deoxynucleotidyl transferase-mediated deoxyuridine 5 triphosphate nick end-labeling) staining and caspase-3 immunohistochemistry. Diabetes was induced in normal rats using streptozotocin and maintained for 5 to 6 weeks. The middle cerebral artery of both normal and diabetic rats was occluded and maintained from 24 or 48 hours. Sham-operated normal and diabetic animals served as controls. Following 24 to 48 hours of occlusion, the animals were sacrificed and the brains were removed, sectioned, and processed for TUNEL staining or caspase-3 immunohistochemistry. Middle cerebral artery occlusion in normal rats was associated with an increase in the number of both TUNEL-positive and caspase-3-positive cells in selected brain regions (hypothalamic preoptic area, piriform cortex, and parietal cortex) when compared to nonoccluded controls. Diabetic rats without occlusion showed significant increases in both TUNEL-positive and caspase-3-positive cells compared to normal controls. Middle cerebral artery occlusion in diabetic rats resulted in increases in TUNEL positive as well as caspase-3-positive cells in selected regions, above those seen in nonoccluded diabetic rats. Both TUNEL staining and caspase-3 immunohistochemistry revealed that the number of apoptotic cells in diabetic animals tended to be greatest in the preoptic area and parietal cortex. The authors conclude that focal cerebral ischemia is associated with a significant increase in apoptosis in nondiabetic rats, and that diabetes alone or diabetes plus focal ischemia are associated with significant increases in apoptotic cells. PMID- 12745667 TI - Effects of postnatal stress on the development of type 1 diabetes in bank voles (Clethrionomys glareolus). AB - Wild bank voles (Clethrionomys glareolus) kept in the laboratory under barren housing conditions develop high incidences of type 1 diabetes mellitus due to beta cell-specific lysis in association with the appearance of GAD65, IA-2, and insulin autoantibodies. Wild-caught and immediately analyzed voles show no histological signs of diabetes, and the disease may therefore be induced by circumstances related to the housing of the animals in captivity. We tested the possibility that postnatal stress by either maternal separation or water immersion at different intervals would induce diabetes in adult bank voles. We found that low-frequent stress during the first 21 days of life increases, whereas high-frequent stress markedly reduces, the incidence of type 1 diabetes in adulthood. These results differentiate the role of early-experienced stress on subsequent type 1 diabetes development and emphasize that the bank vole may serve as a useful new animal model for the disease. PMID- 12745668 TI - A lipoprotein lipase-promoting agent, NO-1886, improves glucose and lipid metabolism in high fat, high sucrose-fed New Zealand white rabbits. AB - The synthetic compound NO-1886 is a lipoprotein lipase activator that lowers plasma triglycerides and elevates high-density lipoprotein cholesterol (HDL-C). Recently, the authors found that NO-1886 also had an action of reducing plasma glucose in high-fat/high-sucrose diet-induced diabetic rabbits. In the current study, we investigated the effects of NO-1886 on insulin resistance and beta-cell function in rabbits. Our results showed that high-fat/high-sucrose feeding increased plasma triglyceride, free fatty acid (FFA), and glucose levels and decreased HDL-C level. This diet also induced insulin resistance and impairment of acute insulin response to glucose loading. Supplementing 1% NO-1886 into the high-fat/high-sucrose diet resulted in decreased plasma triglyceride, FFA, and glucose levels and increased HDL-C level. The authors also found a clear increased glucose clearance and a protected acute insulin response to intravenous glucose loading by NO-1886 supplementation. These data suggest that NO-1886 suppresses the elevation of blood glucose in rabbits induced by feeding a high fat/high-sucrose diet, probably through controlling lipid metabolism and improving insulin resistance. PMID- 12745669 TI - Development of type 1 diabetes in wild bank voles associated with islet autoantibodies and the novel ljungan virus. AB - Wild bank voles (Clethrionomys glareolus) may develop diabetes in laboratory captivity. The aim of this study was to test whether bank voles develop type 1 diabetes in association with Ljungan virus. Two groups of bank voles were analyzed for diabetes, pancreas histology, autoantibodies to glutamic acid decarboxylase (GAD65), IA-2, and insulin by standardized radioligand-binding assays as well as antibodies to in vitro transcribed and translated Ljungan virus antigens. Group A represented 101 trapped bank voles, which were screened for diabetes when euthanized within 24 hours of capture. Group B represented 67 bank voles, which were trapped and kept in the laboratory for 1 month before being euthanized. Group A bank voles did not have diabetes. Bank voles in group B (22/67; 33%) developed diabetes due to specific lysis of pancreatic islet beta cells. Compared to nondiabetic group B bank voles, diabetic animals had increased levels of GAD65 (P < .0001), IA-2 (P < .0001), and insulin (P = .03) autoantibodies. Affected islets stained positive for Ljungan virus, a novel picorna virus isolated from bank voles. Ljungan virus inoculation of nondiabetic wild bank voles induced beta-cell lysis. Compared to group A bank voles, Ljungan virus antibodies were increased in both nondiabetic (P < .0001) and diabetic (P = .0015) group B bank voles. Levels of Ljungan virus antibodies were also increased in young age at onset of newly diagnosed type 1 diabetes in children (P < .01). These findings support the hypothesis that the development of type 1 diabetes in captured wild bank voles is associated with Ljungan virus. It is speculated that bank voles may have a possible zoonotic role as a reservoir and vector for virus that may contribute to the incidence of type 1 diabetes in humans. PMID- 12745670 TI - Des(1-3)IGF-1 treatment normalizes type 1 IGF receptor and phospho-Akt (Thr 308) immunoreactivity in predegenerative retina of diabetic rats. AB - Little is known about interventions that may prevent predegenerative changes in the diabetic retina. This study tested the hypothesis that immediate, systemic treatment with an insulin-like growth factor (IGF)-1 analog can prevent abnormal accumulations of type 1 IGF receptor, and phospho-Akt (Thr 308) immunoreactivity in predegenerative retinas of streptozotocin (STZ) diabetic rats. Type 1 IGF receptor immunoreactivity increased approximately 3-fold in both inner nuclear layer (INL) and ganglion cell layer (GCL) in retinas from STZ rats versus nondiabetic controls. Phospho-Akt (Thr 308) immunoreactivity increased 5-fold in GCL and 8-fold in INL of STZ rat retinas. In all cases, immunoreactive cells were significantly reduced in STZ des(1-3)IGF-1-treated versus STZ rats. Preliminary results suggested that vascular endothelial growth factor (VEGF) levels may also be reduced. Hyperglycemia/failure of weight gain in diabetic rats continued despite systemic des(1-3)IGF-1. These data show that an IGF-1 analog can prevent early retinal biochemical abnormalities implicated in the progression of diabetic retinopathy, despite ongoing hyperglycemia. PMID- 12745671 TI - Omapatrilat, an angiotensin-converting enzyme and neutral endopeptidase inhibitor, attenuates early atherosclerosis in diabetic and in nondiabetic low density lipoprotein receptor-deficient mice. AB - Omapatrilat inhibits both angiotensin-converting enzyme (ACE) and neutral endopeptidase (NEP). ACE inhibitors have been shown to inhibit atherosclerosis in apoE-deficient mice and in several other animal models but failed in low-density lipoprotein (LDL) receptor-deficient mice despite effective inhibition of the renin-angiotensin-aldosterone system. The aim of the present study was to examine the effect of omapatrilat on atherogenesis in diabetic and nondiabetic LDL receptor-deficient mice. LDL receptor-deficient male mice were randomly divided into 4 groups (n = 11 each). Diabetes was induced in 2 groups by low-dose STZ, the other 2 groups served as nondiabetic controls. Omapatrilat (70 mg/kg/day) was administered to one of the diabetic and to one of the nondiabetic groups. The diabetic and the nondiabetic mice were sacrificed after 3 and 5 weeks, respectively. The aortae were examined and the atherosclerotic plaque area was measured. The atherosclerotic plaque area was significantly smaller in the omapatrilat-treated mice, both diabetic and nondiabetic, as compared to nontreated controls. The mean plaque area of omapatrilat-treated nondiabetic mice was 9357 +/- 7293 microm2, versus 71977 +/- 34610 microm2 in the nontreated mice (P = .002). In the diabetic animals, the plaque area was 8887 +/- 5386 microm2 and 23220 +/- 10400 microm2, respectively for treated and nontreated mice (P = .001). Plasma lipids were increased by omapatrilat: Mean plasma cholesterol in treated mice, diabetic and nondiabetic combined, was 39.31 +/- 6.00 mmol/L, versus 33.12 +/- 7.64 mmol/L in the nontreated animals (P = .008). The corresponding combined mean values of triglycerides were 4.83 +/- 1.93 versus 3.00 +/- 1.26 mmol/L (P = .02). Omapatrilat treatment did not affect weight or plasma glucose levels. Treatment with omapatrilat inhibits atherogenesis in diabetic as well as nondiabetic LDL receptor-deficient mice despite an increase in plasma lipids, suggesting a direct effect on the arterial wall. PMID- 12745672 TI - Scanning electron microscopic characterization of healing and normal rat ligament microstructure under slack and loaded conditions. AB - The objective of this study was to observe and compare behavior of the collagen fiber microstructure in normal and healing ligaments, both in situ and ex vivo, in order to add insight into the structure-function relationship in normal and healing ligaments. Fifty-two ligaments from 26 male rats were investigated. Eleven animals underwent surgical transection of both medial collateral ligaments (MCLs) (22 ligaments), which were allowed to heal for a period of 2 weeks. An additional 15 animals (30 ligaments) were used as normals. Ligaments were placed into six groups: Slack (n = 6 control, n = 6 healing), Reference (n = 4 control, n = 4 healing), Loaded (n = 4 control, n = 4 healing), 15 degrees Flexion (n = 4 control, n = 4 healing), 120 degrees Flexion (n = 4 control, n = 4 healing), and Tissue Strain vs. Flexion Angle (n = 8 normals). All ligaments, except those in the Tissue Strain vs. Flexion Angle group, were prepared for scanning electron microscopy. Tissues were harvested, mounted in a load frame, and chemically fixed in one of five states: (1). slack, (2). reference (onset of loading), (3). loaded, (4). 15 degrees knee flexion, or (5). 120 degrees knee flexion. After fixation the tissues were prepared for electron microscopy (SEM). The micrographs from the slack, reference, and loaded groups show fiber straightening with loading in normal ligaments as well as in both scar and "retracted" regions of healing ligaments. Collagen fibers' diameter and crimp patterns were dramatically changed in the scar region of healing ligaments: Width decreased from 19.4 +/- 1.7 microm to 6.5 +/- 2.1 microm (p <.000001), period from 51.4 +/- 15.1 microm to 11.0 +/- 2.4 microm (p <.000001), and amplitude from 9.8 +/- 0.8 microm to 3.9 +/- 0.8 microm (p <.000001). Normal ligaments fixed in situ show wavy regions at 120 degrees but less so at 15 degrees flexion. Healing ligaments fixed in situ show regions of fiber waviness in the scar region at 120 degrees and also at 15 degrees flexion, indicating ligament laxity persists toward both extremes of the range of motion. The data suggest that straightening of crimped fibers is a functionally relevant phenomenon, not only in normal but also in healing ligaments. PMID- 12745673 TI - Temporal and spatial localization of proteoglycan decorin transcripts during the progression of cholesterol-induced atherosclerosis in Japanese quail. AB - The temporal and spatial distribution of decorin transcripts, endothelial cells, and smooth muscle actin-producing cells were determined during the progression of atherosclerosis in the dorsal aortas of Japanese quail selected for cholesterol induced atherosclerosis. The quail were placed on either a control diet or a diet containing 0.5% added cholesterol at approximately 16 weeks of age. Dorsal aortas were collected at 2-week intervals for 18 weeks after initiating cholesterol feeding. In situ hybridization for decorin showed the presence of decorin transcripts in the dorsal aorta intima of cholesterol-fed birds beginning at 6 weeks and continuing through the duration of the study. However, in the control fed birds, decorin transcripts were not found in the intima but were prominent in the adventitia. In cholesterol-fed birds, immunohistochemical localization for endothelial cells showed that intima areas positive for decorin transcripts colocalized with both the endothelial cells and smooth muscle cell layers. In contrast in the control-fed birds, decorin transcripts aligned with the smooth muscle cell layers and not the endothelial cells. These results are suggestive of a potential role for endothelial cells in intimal decorin expression during atherosclerotic plaque formation. PMID- 12745675 TI - Altered expression of matrix metalloproteinases within mineralizing bone cells in vitro in the presence of fluoride. AB - Fluoride is known to alter mineralization within bone, although the mechanism for its action is unclear. An important stage in the formation of mineralized tissues is the remodeling of the osteoid, facilitating mineral deposition. Using a bone mineralizing culture system derived from rat femur washes, this study investigated the influence of fluoride on MMP expression at a developmental stage relating to the onset of mineralization. Bone cells cultured in the absence of fluoride synthesized an active form of a 45-kD MMP, which was immunoreactive with an antibody to human MMP-1 (although full characterization of this MMP was not achieved), trace levels of an MMP immunoreactive with anti-MMP-3, and a 66-kD proteolytic species. Incubation in 10(-7) and 10(-5) M fluoride resulted in a decrease in expression of the 45-kD MMP, sharp increases in the expression of MMP 3, and the appearance of a band at 110 kD, which showed immunoreactivity for MMP 9. The influence of fluoride on MMP expression is likely to influence the composition of the remodeling matrix and subsequent mineralization and offers a potential mechanism by which fluoride alters mineralization. PMID- 12745674 TI - Cultures of ligament fibroblasts in fibrin matrix gel. AB - The cellular properties of anterior cruciate ligament (ACL) and medial collateral ligament (MCL) fibroblasts have been analyzed in a three-dimensional fibrin matrix gel (FMG) system. The MCL fibroblasts proliferated significantly faster than ACL fibroblasts in 10% fetal bovine serum (FBS). FMG contraction resembles soft-tissue wound contraction. Transforming growth factor-beta1 (TGF-beta1) (5 ng/ml) caused a significantly faster rate of FMG contraction than control (0.5% FBS) in both ACL and MCL fibroblasts. Unlike the cells in 10% FBS, this faster rate of FMG contraction was achieved without increasing the initial cell number. In the FMG, the MCL fibroblasts demonstrated significantly higher collagen synthesis per cell than ACL fibroblasts between the days 2 and 6 of culture. These differences in cellular properties of the ACL and MCL fibroblasts that were observed in vitro may explain the differences in the healing potential of these ligaments in vivo. PMID- 12745677 TI - Effect of reinsertion of activated nucleus pulposus on disc degeneration: an experimental study on various types of collagen in degenerative discs. AB - We examined the emergence and sequential changes in type I, II, and VI collagen production in an experimental rabbit model of disc degeneration. Type I collagen was minimally present initially and did not change over 24 weeks. Type I collagen seemed to have no effect on the degenerative process in this model. Staining for type II collagen was positive circumferentially in chondrocytelike cells and was mild in the early phase of disc degeneration, when the chondrocytelike cells began to appear in the inner layers of the annulus fibrosus. The stain became stronger during the middle phase when the chondrocytelike cells arranged themselves in cluster. Compared with type II collagen, the staining for type VI collagen was relatively strong early in the degenerative process. These findings led us to speculate that these chondrocytelike cells play an active role in the degenerative process. The reinsertion of nucleus pulposus cells cocultured with annulus fibrosus delayed disc degeneration and the emergence of chondrocytelike cells. Considering that the emergence of chondrocytelike cells which produce type II and type VI collagen is delayed in discs with the injection of cocultured nucleus pulposus cells by annulus fibrosus cells, we conclude that chondrocytelike cells that produce type VI collagen also seems to accelerate degeneration. Type VI collagen is produced at an earlier phase than type II collagen and may be both active agent and a marker for disc degeneration. PMID- 12745676 TI - Increase of interstitial collagen in the mouse endometrium during decidualization. AB - Decidualization in the mouse consists of an extensive remodeling of the endometrial extracellular matrix, resulting in a reduction of the extracellular spaces, an increase in the diameter of collagen fibrils, and changes in the relative ratio of different types of glycosaminoglycans. To assess the dynamic changes of the endometrial extracellular matrix during decidualization, collagen was analyzed biochemically and immunochemically in the endometrium of nulliparous and day 5 to day 8 pregnant mice. The amount of collagen per gram dry weight was higher in the endometrium of implantation sites than in interimplantation sites. Collagen types I, III, and V were the main components of the endometrium of nulliparous and pregnant animals. The amount of collagen type V was higher in the endometrium of pregnant animals than in nulliparous ones. A relative unusual homotrimeric form of collagen type V, probably formed by [alpha1(V)](3), was detected in pregnant endometrium by gel eletrophoresis and immunoblotting. PMID- 12745678 TI - A qualitative analysis of crack propagation in articular cartilage at varying rates of tensile loading. AB - A custom-built miniature tensile testing apparatus was used to study the propagation of cracks through the articular cartilage matrix at various loading rates and initial crack lengths. The crack propagation mechanism was observed to be significantly dissimilar to that normally seen in traditional fracture mechanics opening mode, where fracture propagates through the thickness of samples or perpendicularly to the applied load. Instead, an artificially initiated microcrack in the surface layer of an articular cartilage sample grew laterally in the direction of the applied load, stretching about the crack tip, whose initial position remained unchanged throughout the fracture process. A progressive upward pull of the bottom layer toward the surface, which resulted in necking of the specimen, was observed. Our analysis revealed that the rate of necking was the same as that of the lateral stretch of the growing crack. We hypothesize that necking is due to the response of the collagen meshwork especially in the deep zones of the matrix to the tensile load. Our samples exhibited unstable fracture growth immediately after each microcrack grew to the base of the articular surface layer, with very fast crack propagation to failure, thereby indicating that the fracture toughness of the articular cartilage matrix is significantly determined by the toughness of its articular surface. PMID- 12745679 TI - The night float paradigm to decrease sleep deprivation: good solution or a new problem? AB - In the late 1980s physician residency training programs developed the night float rotation, characterized by a sequence of 5 - 15 days of night work without any daytime duties, thereby involving an abrupt reversal of the wake - sleep schedule. We examined the effect of the night float rotation on sleep, mood and performance of pediatric residents. Residents completed sleep diaries daily, and tests of mood (Profile of Mood States) and attention (Conner's Continuous Performance Test) three times a week during the two-week night float rotation, and during equivalent blocks of time of their daytime rotations. Results show that, despite having ample opportunity to sleep during the day, while on night float rotation residents slept less than during the nights of their normal daytime rotations, 6.3 h +/- 2.5 h and 7.2 h +/- 1.7 h, respectively, p < 0.0001. Also, during night float compared to daytime rotations residents had increased fatigue-inertia scores, 8.7 +/- 4.1 and 4.8 +/- 2.4, respectively, p < 0.0001, and decreased vigor-activity scores 10.7 +/- 5.4 and 14.8 +/- 5.3, respectively, p = 0.02. The scores for attention were not significantly different between night float and daytime rotations. The correlation coefficients of fatigue with measures of attention were not statistically significant for daytime rotations. However, for night float fatigue correlated with omission errors, r = 0.51, p = 0.001 and with attentiveness r = - 0.36, p = 0.03. Training programs that adopt the night float rotation must be aware of potential deleterious effects of the night float rotation as they may lead to serious consequences on residents' performance and patients' safety. PMID- 12745680 TI - Need for recovery from work: evaluating short-term effects of working hours, patterns and schedules. AB - In this paper working hours, patterns and work schedules of employees were evaluated in terms of need for recovery from work. Self-administered questionnaire data from employees of the Maastricht Cohort Study on Fatigue at Work (n = 12,095) were used. Poisson regression analyses and multivariate logistic regression analyses revealed that higher working hours a day and working hours a week generally went together with more need for recovery from work. Overtime work was particularly associated with higher need for recovery from work in both genders. Both male and female three-shift or irregular shift workers had higher odds of elevated need for recovery compared to day workers. When additionally controlling for work-related factors, need for recovery levels among shift workers substantially lowered. This study clearly showed that working hours and schedules are associated with need for recovery from work, with different associations for men and women. Especially the associations between work schedules and need for recovery from work were very interrelated with other work related factors. Future studies could further investigate the possibility that shift work might function as a proxy of other work-related factors that explain the different levels in need for recovery from work, or that job demands are perceived higher among shift workers and may therefore lead to more need for recovery from work. PMID- 12745681 TI - Revision of the design of a standard for the dimensions of school furniture. AB - In this study an anthropometric design process was followed. The aim was to improve the fit of school furniture sizes for European children. It was demonstrated statistically that the draft of a European standard does not cover the target population. No literature on design criteria for sizes exists, and in practice it is common to calculate the fit for only the mean values (P50). The calculations reported here used body dimensions of Dutch children, measured by the authors' Department, and used data from German and British national standards. A design process was followed that contains several steps, including: Target group, Anthropometric model and Percentage exclusion. The criteria developed in this study are (1) a fit on the basis of 1% exclusion (P1 or P99), and (2) a prescription based on popliteal height. Based on this new approach it was concluded that prescription of a set size should be based on popliteal height rather than body height. The drafted standard, Pren 1729, can be improved with this approach. A European standard for school furniture should include the exception that for Dutch children an extra large size is required. PMID- 12745682 TI - The identification of knowledge content and function in manual labour. AB - Calls for an alternative conceptualization of cognition for applied concerns retain the core commitment of the basic research community to abstract cognition detached from a physical environment. The present paper attempts to break out of the dominant, narrow view of cognition and cognitive domains, with a cognitive analysis of digging ditches for the utility industry. To illustrate knowledge based cognition in manual labour excerpts are presented from the journal entries of a moderately experienced student working a summer job, organized with a representation that distinguishes between the goals and methods of work. The journal entries illustrate the functions of knowledge for interacting with a physical environment; knowledge enables the selection, execution and monitoring of work methods, the interpretation of perceptual information, the application of task completion criteria and the ability for explanation and generalization. To emphasize the generality of the functions of cognition in ditch digging, comparable functions are indicated in a domain rarely regarded as a form of manual labour: the practice of internal medicine. Discussion of the results includes the implications for cognitive theory as well as practical implications for productivity, training and task analysis. PMID- 12745683 TI - Posture, muscle activity and muscle fatigue in prolonged VDT work at different screen height settings. AB - With the increasing use of video display terminals (VDTs), there is growing concern over the corresponding increase in the number of health problems reported. Although much research has focused on identifying the optimal screen height, there is to date no consensus. This study aimed to investigate the effect of prolonged (89 min) VDT work at four different screen heights on head-neck posture, muscle activity and the development of muscle fatigue. The results show that lowering screen height, starting from 15 cm above the baseline (i.e. top of the screen level with eye height while sitting), decreased the ear-eye angle, increased the viewing angle, increased the viewing angle relative to the ear-eye line, and increased the muscle activity of the neck extensor muscles. There were also some significant time effects on postural angles and muscle activity. In this study there were only rare occurrences of muscle fatigue, defined as a simultaneous increase in EMG amplitude and a shift of the EMG power spectrum to lower frequencies. Muscle activity increased significantly in some muscles and for certain screen heights. PMID- 12745684 TI - Previous history of LBP with work loss is related to lingering deficits in biomechanical, physiological, personal, psychosocial and motor control characteristics. AB - A cross-sectional retrospective study was made of currently asymptomatic workers who perform physically demanding jobs. To further quantify the association between various biomechanical, physiological, personal psychosocial and motor control parameters that linger due to a history of low back disorders. Seventy two workers were recruited from heavy industry, 26 of whom had a history of disabling low back disorders (LBDs) sufficient to miss work while the others did not. The strength of the study lies in the many detailed variables measured. Having a history of low back disorders was found to be associated with a larger waist girth, a greater potential for low back pain chronicity as predicted from psychosocial questionnaires, perturbed flexion to extension strength and endurance ratios, and widespread motor control deficits across a variety of tasks, some of which resulted in high back loads. In those workers who had missed work due to back disorders, the length of time since their last disabling episode was 261 weeks on average, suggesting that multiple deficits may remain for a period of time. Having a history of LBD is associated with changes in attitudes, in body composition, and in the way people move, load their backs and respond to a variety of motor and stability challenges. PMID- 12745685 TI - The use of continuous exposure data for predicting CTS in fish processing operators. AB - Carpal tunnel syndrome (CTS) remains one of the most commonly reported and studied work related musculoskeletal disorders. Categorical representations of exposures has been critical in identifying associations between risk factors and CTS, however, quantification of exposure-response relationships require using continuous exposure data. Also, few interactions between risk factors, especially between risk factor categories, have been investigated. The objectives of this study were to investigate the utility of using continuous exposure data and to identify interaction effects of risk factors, both within and between risk factor categories, for predicting CTS. A cross sectional study was performed at a fish processing facility in which 53 participants were evaluated during normal task performance. Due to task asymmetry, each hand was considered separately, providing 106 hands for analysis. Direct measurement and a questionnaire were used to quantify exposures to common occupational and personal risk factors. Stepwise logistic regression analysis was performed to identify three models for predicting CTS and assess predictive ability using: occupational risk factors only (three-way interactions considered), personal risk factors only (two-way interactions considered), and a mixed model considering two-way interactions across risk factor categories and previously identified significant interactions. Models including only occupational or personal risk factors were moderately accurate overall (73% and 77% respectively), but were not sensitive in differentiating between CTS cases and non-cases (39% and 33% respectively). The mixed model was found to be accurate (88%) and sensitive (78%), though only one interaction effect was included. The results of this study illustrate the importance of using continuous exposure data, especially in job tasks where exposures to occupational risk factors is similar, when differentiating between high and low risk job tasks. PMID- 12745687 TI - Effect of a redesigned two-wheeled container for refuse collecting on mechanical loading of low back and shoulders. AB - The objective of this study was to compare the mechanical and perceived workload when working with a redesigned two-wheeled container and working with a standard two-wheeled container for refuse collecting. The three changes in the design of the container were a displacement of the position of the centre of mass in the direction of the axis of the wheels, a slight increase in the height of the handle and a slight increase in the horizontal distance between the handle and the wheel-axis, and an increase in the diameter of the wheels. The volume of the container remained 0.240 m3. Nine refuse collectors performed some of their most frequent daily activities with both types of containers in the laboratory. Kinematics and exerted hand forces were assessed as input for detailed 3D biomechanical models of the low back and shoulder to estimate net moments at the low back and shoulders, compressive forces at the low back and contact forces at the glenohumeral joint. Also, the refuse collectors rated the ease of handling the two-wheeled containers on a five point scale. The use of the redesigned container resulted in a decrease of the exerted hand forces of 27%, decreases in the net moments at the low back and shoulders of 8% and 20%, respectively, and a decrease of 32% of the contact force at the glenohumeral joint when compared to the standard container. However, pulling an empty redesigned container on to the pavement resulted in an increase of the shoulder moment of more than 100%. No differences between container types were found for the compressive forces at the low back. Pushing and pulling with the redesigned container was rated as easier than pushing and pulling with the standard container. No differences in subjective ratings were found for the tasks of turning the container or pulling an empty container onto the pavement. It is concluded that, provided that empty containers are placed back onto the pavement as infrequently as possible, the introduction of the redesigned container could result in a reduction of the low back and shoulder load for refuse collectors. PMID- 12745688 TI - Effects of chronic low back pain, age and gender on vertical spinal creep. AB - This study investigated the effect of chronic low back pain, age, gender, and time of measurement on the magnitude of vertical spinal creep (VSC) and its recovery. A mixed design, involving three independent variables (chronic low back pain, age, and gender) and one repeated variable (time), was used. One hundred and six subjects of both genders, with and without chronic low back pain, aged between 20 and 60 years, participated in the study. The measurement of VSC and its recovery was performed using a seated stadiometer that allowed continuous measurement of VSC without changing the subject's posture over 25 min. Unloaded VSC was measured during the initial 5 min, followed by 10 min with an additional load of 15% of the subject's body weight and then for a further 10 min after the removal of the load. Subjects were grouped into one of eight categories according to the presence of chronic low back pain, age (20-39 years or 40-60 years) and gender. Repeated measures analysis of variance was computed. A significant increase in VSC with time of measurement was observed (p<0.001). No significant main effects for chronic low back pain, age, or gender were found at any time during the 25-min VSC testing protocol. Significant interactions were found between age and gender during the loaded (p=0.02) and unloaded (p=0.02) phases. A significant interaction was found between chronic low back pain and gender at the end of the unloaded phase (p=0.04). These findings suggest a combined influence of chronic low back pain, age, and gender on VSC and its recovery and that the dominance of each variable changed with the time of the measurement. Thus, subjects who differ in the presence of chronic low back pain, age, and gender should not be combined for statistical analysis of VSC and its recovery. PMID- 12745689 TI - What is a demanding lifting job for manual handling workers in Hong Kong? AB - Although lifting tasks are traditionally evaluated by researchers, through the use of methods which depend on one or more approaches (i.e., psychophysical, biomechanical and physiological methods), none of these approaches makes use of expert workers in the evaluation of lifting activities. Given that 97% of lower back symptoms are aches, pains and discomfort and rely on self-reports, it is intuitive to use expert workers in evaluating the stressfulness of lifting activities. In this investigation, 217 workers from three industries in the Hong Kong area were surveyed to determine what constitutes a demanding lifting job from a worker standpoint. This was achieved by asking workers to map, in numerical terms, the level of lifting task parameter described in linguistic values. For example, the weight of load was described in three linguistic descriptors, namely, 'light', 'moderate' and 'heavy'. Then, each worker was asked to assess the meaning of these linguistic descriptors in numerical terms, based on their perception and experience with manual handling work. In this study, workers were asked to map the physical analogue of linguistic descriptors for seven lifting task parameters which are utilized in the NIOSH lifting equation, that is, weight of load, horizontal distance, vertical travel distance, vertical origin of lift, angle of asymmetry, frequency of handling and task duration. The data obtained from the workers were then subjected to validity testing in relation to norms and values reported in the published literature. On a preliminary basis, it appears that workers' assessments are logical and valid. The results suggest that the worker-based methodology is a promising approach and that it is worthwhile to pursue this approach further in the evaluation of industrial lifting activities. PMID- 12745690 TI - The impact of working technique on physical loads - an exposure profile among newspaper editors. AB - The aim of this study was to investigate the possible associations between working technique, sex, symptoms and level of physical load in VDU-work. A study group of 32 employees in the editing department of a daily newspaper answered a questionnaire, about physical working conditions and symptoms from the neck and the upper extremities. Muscular load, wrist positions and computer mouse forces were measured. Working technique was assessed from an observation protocol for computer work. In addition ratings of perceived exertion and overall comfort were collected. The results showed that subjects classified as having a good working technique worked with less muscular load in the forearm (extensor carpi ulnaris p=0.03) and in the trapezius muscle on the mouse operating side (p=0.02) compared to subjects classified as having a poor working technique. Moreover there were no differences in gap frequency (number of episodes when muscle activity is below 2.5% of a reference contraction) or muscular rest (total duration of gaps) between the two working technique groups. Women in this study used more force (mean force p=0.006, peak force p=0.02) expressed as % MVC than the men when operating the computer mouse. No major differences were shown in muscular load, wrist postures, perceived exertion or perceived comfort between men and women or between cases and symptom free subjects. In conclusion a good working technique was associated with reduced muscular load in the forearm muscles and in the trapezius muscle on the mouse operating side. Moreover women used more force (mean force and peak force) than men when operating the click button (left button) of the computer mouse. PMID- 12745691 TI - Environmental and human factors influencing thermal comfort of office occupants in hot - humid and hot - arid climates. AB - The effects of environmental and individual factors on thermal sensation in air conditioned office environments were analysed for two large, fully compatible thermal comfort field studies in contrasting Australian climates. In the hot - humid location of Townsville, 836 office workers were surveyed; 935 workers participated in hot - arid Kalgoorlie-Boulder. Overall perceived work area temperature and measured indoor operative temperature correlated moderately with thermal sensation for Townsville (T) subjects but only perceived temperature correlated with Kalgoorlie-Boulder (KB) sensation. Multiple regression analyses confirmed that indoor climatic variables (including Predicted Mean Vote) contributed to actual thermal sensation vote (24% T; 15% KB), with operative temperature having more of an effect in T than in KB. Subsequent analyses of individual characteristics showed no linear contributions to thermal sensation. The remaining variances were significantly related to perceived work area temperature (7% additional explained variance in T; 12% in KB). Mann - Whitney analyses (after correction for climatic variables) showed that T subjects with higher job satisfaction had thermal sensations closer to 'neutral'. Males, healthier subjects, non-smokers, respondents with earlier survey times and underweight occupants had lower median thermal sensations in KB. Townsville occupants appeared more adapted to their outdoor climatic conditions than Kalgoorlie-Boulder respondents, perhaps due to limited home air-conditioning. Further research into non-thermal impacts on gender-related thermal acceptability is suggested. PMID- 12745692 TI - Visual attention while driving: sequences of eye fixations made by experienced and novice drivers. AB - Eye fixations were recorded while novice and experienced drivers drove along three types of roads (rural, suburban and dual-carriageway). An analysis of the content of those fixations was performed in order to identify differences in the scanpaths that can be associated with skill acquisition and that can indicate a sensitivity to road type. This analysis itemized the part of the visual scene that was inspected with each fixation, and identified what the driver looked at as a function of what they had looked at previously. Single-fixation, two fixation, and three-fixation patterns of eye-movements were identified. Differences in sequences of fixations were found between novice and experienced drivers on the three types of roads, with experienced drivers showing greater sensitivity overall, and with some stereotypical transitions in the visual attention of the novices. A number of individual sequences were identified, including a roadway preview pattern (alternating fixations between near and far views of the road ahead), and patterns involving mirror inspections that varied according to the road type. PMID- 12745694 TI - Criteria for driver impairment. AB - Most traffic accidents can be attributed to driver impairment, e.g. inattention, fatigue, intoxication, etc. It is now technically feasible to monitor and diagnose driver behaviour with respect to impairment with the aid of a limited number of in-vehicle sensors. However, a valid framework for the evaluation of driver impairment is still lacking. To provide an acceptable definition of driver impairment, a method to assess absolute and relative criteria was proposed to fulfil the paradoxical goal of defining impaired driving which is consistent yet adaptable to interindividual differences. PMID- 12745695 TI - An investigation of stability limits while holding a load. AB - The objective of this study was to investigate the effects of load height and foot placement on the functional base of support (FBOS) limits and the postures that participants used when they reached their FBOS limits. Twelve young male participants were tested while holding a 12-kg load at reach (above their heads), at shoulder and knuckle height, and unladen under both wide and narrow foot placements. The FBOS limits and the centre of pressure (COP) excursion length were calculated based on data from a force platform. Postural angles when participants reached their FBOS limits were calculated from records of a 2-D motion analysis system. The results showed that the load height had greater effect on the posterior FBOS limit. As the load height decreased, the COP excursion length decreased. Participants were prone to using a hip strategy to maintain postural balance when reaching their FBOS limits. Quantitative data of FBOS limits and postural control while participants hold a load at various heights when reaching their FBOS limits is of value for designing a safe materials handling workplace. PMID- 12745696 TI - The effect of work-rest schedules and type of task on the discomfort and performance of VDT users. AB - Consideration of the literature survey indicates that video display terminal (VDT) operators tend to have a high incidence of musculoskeletal problems, visual fatigue, and job stress. Although a number of ergonomic improvements in workstation design and work environment can help to reduce these problems, a proper work-rest schedule deserves consideration since it is easily applicable and inexpensive. The objective of this study was to compare the work-rest schedules for VDT operators considering data entry and mental arithmetic tasks. An experiment was conducted with 10 male college students as participants. The methodology included a discomfort questionnaire and performance measures. The independent variables were the work-rest schedule (60-minute work/10-minute rest, 30-minute work/5-minute rest, and 15-minute work/micro breaks) and the type of task (data entry and a mental arithmetic task). The results were analysed using multiple analysis of variance followed by separate analyses. The 15/micro schedule resulted in significantly lower discomfort in the neck, lower back, and chest than the other schedules for data entry task. The 30/5 schedule followed by 15/micro schedule resulted in the lowest eyestrain and blurred vision. Discomfort in the elbow and arm was the lowest with the 15/micro schedule for the mental arithmetic task. The 15/micro schedule resulted in the highest speed, accuracy, and performance for both of the tasks, compared with the 60/10 and 30/5 schedules. The data entry task resulted in significantly increased speed, accuracy, and performance, and lower shoulder and chest discomfort than the mental arithmetic task. PMID- 12745697 TI - A strategy for improving worker satisfaction and job attitudes in a repetitive industrial task: application of production standards and performance feedback. AB - Worker satisfaction improved significantly as a consequence of the provision of the assigned and participative standards with performance feedback in a repetitive industrial production task. The maximum improvement in worker satisfaction was found for the participative standard and feedback condition. Only this condition had a significant positive effect on worker job attitudes. Monetary incentive, when provided with an assigned or participative standard with feedback, added no incremental worker satisfaction or job attitudes gain. The participative standard with feedback condition emerges as the optimum strategy for improving worker satisfaction and job attitudes in a repetitive industrial production task. PMID- 12745698 TI - Cognitive flexibility and adaptability to environmental changes in dynamic complex problem-solving tasks. AB - People who show good performance in dynamic complex problem-solving tasks can also make errors. Theories of human error fail to fully explain when and why good performers err. Some theories would predict that these errors are to some extent the consequence of the difficulties that people have in adapting to new and unexpected environmental conditions. However, such theories cannot explain why some new conditions lead to error, while others do not. There are also some theories that defend the notion that good performers are more cognitively flexible and better able to adapt to new environmental conditions. However, the fact is that they sometimes make errors when they face those new conditions. This paper describes one experiment and a research methodology designed to test the hypothesis that when people use a problem-solving strategy, their performance is only affected by those conditions which are relevant to that particular strategy. This hypothesis is derived from theories that explain human performance based on the interaction between cognitive mechanisms and environment. PMID- 12745699 TI - A biomechanical analysis of loads on x-ray technologists: a field study. AB - The aim of the study was to determine the biomechanical loads on radiographers in their occupation. Seven x-ray technologists (one male and six females with combined mean age 32.5 years, mean height 164.6 cm and mean weight 68.3 kg) volunteered to be videotaped while doing their jobs (same as normal and others in a simulated manner). X-ray technologists perform all these tasks regularly every day as part of their assignments. The magnitude of the load handled was measured on weighing scale and recorded. The videotape was played back in the laboratory and the initial and final frames of the tasks investigated were frozen for analysis. The joint angles, the height and weight of the technologist, and the load on their hands (weighed before the task was performed) were input into the 3 D Michigan static strength model for calculation of the lumbosacral load and loads on the joints of the extremities. The lumbosacral compression and shear loads were calculated for 16 tasks commonly performed by all x-ray technologists. The x-ray technologists' work was found to be biomechanically quite demanding. Tasks such as repositioning patients horizontally and lifting a patient from a wheelchair caused lumbosacral compression loads of 7,936N and 8,335N respectively, in which exceeded the maximum permissible limit set by NIOSH in 1981. The action limit defined by NIOSH was exceeded by other tasks. It is also of interest that the majority of the x-ray technologists were female. PMID- 12745700 TI - Free shoulder space requirements in the design of high backrests. AB - The objective of this study was to determine the influence of scapular support on the effects of lumbar support and to prove that a high and straight backrest is inappropriate. In literature the importance of a lumbar support is noted, although data about optimal dimensions is an under-researched topic and in earlier studies on force distribution and muscle activity the backrest had a fixed form. The lumbar support is needed to maintain the lumbar lordosis but no studies deal with the question of the precise dimensions of the backrest at shoulder level. With a specially designed apparatus, forces on shoulder and seat were measured separately, and the force on the pelvis calculated, while varying seat and backrest inclination within the range from 0 degrees to 17 degrees. Seat to-backrest angle (at the level of lumbar support) was kept constant at 90 degrees. The distance between the tangent to the lumbar support and the parallel tangent to the scapular support was varied from 0, 2, 4, 6 and 8 cm. This distance is called the free shoulder space. Electromyography was measured at the erector spinae at the levels of the L1, T8 and T5 vertebrae. For all seat angles, a free shoulder space of d=0 cm resulted in the highest back muscle activity. In agreement with the biomechanical model, EMG activity reduced with an increase of seat tilt and increase of free shoulder space. With increasing free shoulder space, a larger part of the total backrest force was carried by the lumbar support. This study shows that a high and straight backrest overrules lumbar support. Offering free shoulder space of at least 6 cm reduces back muscle activity and allows for lumbar support. PMID- 12745702 TI - Long-term adjustment of families following traumatic brain injury where comprehensive rehabilitation has been provided. AB - OBJECTIVES: The present study aimed to examine long-term family and emotional adjustment in close relatives of individuals with TBI, who had access to comprehensive rehabilitation services. It also examined the relative influence thereon of factors including injury severity, handicap and cognitive and behavioural changes in the injured person, relationship with the injured person and caregiver status. METHODS: Participants were 143 TBI individuals and their close relatives. They completed the Family Assessment Device (FAD), Leeds Scales of Anxiety and Depression, Structured Outcome Questionnaire, CHART, SIP Psychosocial Dimension and Novaco Anger Control Questionnaire 2-5 years post injury. RESULTS: Results showed that families were, on average, functioning in the normal range on the FAD. Anxiety and depression were more likely to be present in those responsible for care of their injured relative. There were no differences between spouses and parents. Presence of cognitive, behavioural and emotional changes was the strongest predictor of anxiety and depression in relatives and of unhealthy family functioning. CONCLUSIONS: Every attempt should be made to develop models of long-term support and care that alleviate these sources of burden on relatives. PMID- 12745703 TI - Urinary incontinence after traumatic brain injury: incidence, outcomes and correlates. AB - OBJECTIVES: To document the incidence and outcome of urinary incontinence (UI) and urinary retention (UR) during acute traumatic brain injury (TBI) rehabilitation and determine associated clinical variables. METHODS: Retrospective case series. RESULTS: Eighty-four patients (mean age 44.7 +/- 17.9 years; 66 males, 18 females) with first-ever TBI within 6 weeks of injury. On admission, UI was present in 52 (62%). UR defined as post-void residual urine volumes of > 100 mls was present in eight (9.5%). Forty-six (54.8%) needed external collecting devices and six (7.1%) used indwelling catheters. Bilateral lesions, diabetes mellitus and faecal impaction were present in 70.2, 10.7 and 6.0%, respectively. UI was associated with poorer functional status (measured by the modified Barthel index) and presence of bilateral lesions (p < 0.01). UR was associated with diabetes mellitus and faecal impaction (p < 0.01). On discharge, 31 (36.9%) remained incontinent. CONCLUSION: UI is common after acute TBI and is associated with poorer functional outcome. PMID- 12745704 TI - Relationships between olfactory discrimination and head injury severity. AB - The goal of this study was to examine the relationship between brain injury severity and scores on both an olfactory identification test and on many widely used neuropsychological tests in 367 patients with head injuries of varying levels of severity. It was hypothesized that valid olfactory test scores would correlate highly with injury severity because both the olfactory nerves and the primary olfactory cortices are especially vulnerable to damage in closed head injury. After removing data of doubtful validity from cases failing effort tests, olfactory test scores were related to Glasgow Coma Scale scores (GCS), post traumatic amnesia and radiological abnormalities more strongly than any of the neuropsychological test scores. Based on the assumption that post-traumatic amnesia is caused by a different mechanism than loss of core consciousness, it was also predicted that there would be no cases with a GCS less than 13 and with no post-traumatic amnesia. As predicted, there were no cases in this group. The results support previous studies showing greater olfactory impairment with increased severity of head injury. PMID- 12745705 TI - The use of dopamine enhancing medications with children in low response states following brain injury. AB - PRIMARY OBJECTIVE: The study examines the possible relationship between dopamine enhancing medications and improvement of arousal and awareness in children during persistent low response states (Rancho Los Amigos Levels I, II and III). RESEARCH DESIGN: A retrospective review was conducted of 10 children enrolled in an existing clinical protocol. The Kluge Children's Rehabilitation Center (KCRC) low response protocol provides a double baseline serial measure (A, A, B, B, B) design. Scores on the Western NeuroSensory Stimulation Profile (WNSSP) are the dependent variable. METHODS AND PROCEDURES: Ten children, mean age of 13.7 years low response state (30 days or more) who were treated with dopamine agonists. Co morbid or iatrogenic influences were addressed or ruled out. Seven children had traumatic brain injury, one cerebral vascular accident, one anoxia and one encephalitis. EXPERIMENTAL INTERVENTION: On average, dopamine medications were started 52.9 days post-event. MAIN OUTCOMES AND RESULTS: Paired t-test of WNSSP scores before medications and on medications were significant at p = 0.03 (paired t-test). Also, the distributions of the slopes (rates of change of WNSSP scores over time) were significantly different in the pre-medication and medication phases (Paired T-test, p = 0.02). Random coefficient model comparison of individuals during pre- and medication phase response variability on WNSSP yielded F-test at p = 0.02. CONCLUSIONS: These results suggest a promising relationship between acceleration of recovery for some children in a low response state and administration of dopamine-enhancing medications. PMID- 12745706 TI - Profiles of nursing home residents with traumatic brain injury using the Minimum Data Set. AB - OBJECTIVE: To present comprehensive profiles of nursing home residents with TBI at admission. METHODS: Over 12,300 assessments from the Minimum Data Set were analysed to create these profiles. RESULTS: Residents with TBI were overwhelmingly male and averaged 53.1 years of age at admission, with almost 30% being 40 years or younger. They tended to have substantial physical disability and severe cognitive impairment at admission. Compared to other residents at admission, residents with TBI tended to receive more treatments and rehabilitation therapies. CONCLUSION: Caring for residents with TBI presents major challenges for the staff in the traditional nursing home. PMID- 12745707 TI - Emotional adjustment following cognitive recovery from 'persistent vegetative state': psychological and personal perspectives. AB - Previously, the cognitive recovery of a 26 year old woman, Kate, who developed a severe encephalomyelopathy and was in a 'minimally conscious/persistent vegetative state' for 6 months was reported. After 6 months, Kate began to respond to her environment and, at 2 years post-illness, neuropsychological assessment indicated that Kate was functioning within the normal range on tests of general intellectual functioning, executive functioning and most memory functions (with the exception of visual recognition memory). Although Kate has a severe dysarthria necessitating the use of a communication board and severe physical disabilities that require her to use a wheelchair, she has demonstrated an almost complete cognitive recovery and is among a tiny percentage of minimally conscious patients to do so. This single case report describes the emotional factors central to Kate's rehabilitation. Using a newly developed model of cognitive rehabilitation as a framework, the pivotal role that emotional and psychological factors played in Kate's adjustment to the consequences of her illness and the role of psychotherapeutic intervention in facilitating this adjustment are discussed. PMID- 12745708 TI - Acquired heterotopic ossification in the settings of cerebral anoxia and alternative therapy: two cases. AB - Acquired Heterotopic Ossification (HO) has been well described in the literature as a recognized complication following spinal cord injury, traumatic brain injury and joint arthroplasty. Commonly, large proximal limb joints are affected. The underlying mechanisms for ectopic bone formation remain poorly elucidated. Post stroke hemiplegia as a cause of neurogenic HO is rare, and no published reports of HO occurring after anoxic brain injury in adults have been documented. This study reports two unusual cases of acquired HO: (1) Polyarticular HO involving the ankle joint in a 24-year-old Chinese female who suffered severe anoxic encephalopathy following near drowning which resulted in persistent vegetative state; and (2) Elbow HO in chronic post-stroke hemiplegia occurring as a complication of alternative therapy following repeated forceful manipulation by a traditional practitioner in a 46 year-old male. PMID- 12745709 TI - Crisis and its assessment after brain injury. AB - PRIMARY OBJECTIVE: To develop a measure to assess crisis after acquired brain injury (ABI). RESEARCH DESIGN: A triangulated research strategy, using both qualitative and quantitative methods, was employed to develop the crisis measure. METHODS AND PROCEDURES: The measure was developed in two phases. In the first phase, by using focus group methodology, the experience of crisis following brain injury was described. The second phase involved developing the questionnaire items, pilot testing the measure and conducting initial reliability testing. MAIN OUTCOMES AND RESULTS: The six themes derived from the content analysis led to the creation of the measure, with versions for individuals who have an ABI, family members and professionals. Test-re-test reliability results (n = 40) were adequate. CONCLUSIONS: The results suggest that crisis is experienced as precarious homeostasis with individuals with brain injury, varying in intensity over time, subjectively viewed as never really absent. PMID- 12745710 TI - The mediating effects of effort upon the relationship between head injury severity and cognitive functioning. AB - PRIMARY OBJECTIVE: Given that poor effort may have a profound impact upon the results of neuropsychological assessment, it is of critical importance that assessment of effort is incorporated into the interpretation of a patient's neuropsychological profile. The current study examines the relationship between head injury severity (as measured by length of post-traumatic amnesia) and level of cognitive functioning in patients passing or failing a test of effort (Test of Malingered Memory). RESEARCH DESIGN: A cross-sectional correlational design was employed in a consecutive cohort of 78 persons with ongoing compensation claims. METHODS AND PROCEDURES: Head injury severity was assessed by duration of post traumatic amnesia. All participants received a clinical interview followed by neuropsychological assessment. MAIN OUTCOMES AND RESULTS: A systematic and interpretable negative correlation between head injury severity and intellectual and memory functioning was demonstrated in persons passing the test of effort. However, in persons failing the test of effort no such relationships were demonstrated. CONCLUSIONS: The implications for the interpretation of neuropsychological test data are discussed. It is strongly recommend that valid interpretation of neuropsychological assessment data should include consideration of at least one test of effort. PMID- 12745711 TI - Feasibility of swallowing interventions for tracheostomized individuals with severely disordered consciousness following traumatic brain injury. AB - PRIMARY OBJECTIVE: To report the ability of 12 tracheostomized acute rehabilitation hospital inpatients with severely disordered consciousness post traumatic brain injury (TBI) to participate in an objective swallowing assessment. RESEARCH DESIGN: Post hoc analysis of data from a larger, prospective blinded comparison study. METHODS AND PROCEDURES: Subjects completed a modified barium swallow (MBS) study. Food/drink and tracheostomy tube management recommendations were made. MAIN OUTCOMES AND RESULTS: All subjects participated successfully during an MBS. Post-MBS, 10 subjects began receiving small amounts of food and/or drink. Prior to hospital discharge, all subjects received some food and/or drink and were extubated. Subjects were deemed representative of this patient population and, from a swallowing perspective, other tracheostomized patient populations at the same facility. CONCLUSIONS: Clinicians should routinely consider tracheostomized, acute rehabilitation hospital inpatients with severely disordered consciousness post-TBI potential MBS candidates. Implications and continued research needs are discussed. PMID- 12745712 TI - Clinician expectations for portable electronic devices as cognitive-behavioural orthoses in traumatic brain injury rehabilitation. AB - OBJECTIVE: To ascertain experiences and expectations of clinicians as to uses of portable electronic devices such as hand-held computers as cognitive-behavioural orthoses in TBI rehabilitation. DESIGN: Survey study. PROCEDURES: A survey developed and piloted with the input of clinician focus groups was completed by 81 TBI clinicians from a variety of disciplines and work settings. RESULTS: About half of the respondents reported prior exposure to clients with TBI who had used portable electronic aids. Of 10 key cognitive/behavioural areas, respondents saw most potential for devices in the areas of learning/memory, planning/organization and initiation; less potential was seen for social/interpersonal or behavioural difficulties. However, many respondents also identified learning and memory abilities as essential client characteristics for successful use of portable electronics. Cost was identified as a significant barrier to use of portable devices in rehabilitation. Respondents expressed low confidence overall in their ability to guide clients in use of devices; those who reported personal use were significantly more confident than those who did not. CONCLUSION: Clinicians believe that new electronic devices have potential for treatment of some cognitive impairments, but clinical experimentation may be limited by costs of technology and low clinician confidence. PMID- 12745713 TI - Multisensory integration after traumatic brain injury: a reaction time study between pairings of vision, touch and audition. AB - Traumatic brain injury (TBI) frequently results in deficits in attention and speed of information processing. In order to disentangle the influence of sensory specific factors and the role of cross-modal integration from the supra-modal aspects of cognitive slowing, the present reaction time (RT) study was designed. Simple and choice RT to pairings of visual, auditory and tactile stimuli were measured in 35 TBI patients and 35 matched controls. Results proved a strong influence of sensory-specific and cross-modal factors in the RTs. The tactile modality was more difficult to integrate with the visual and the auditory modality, rather than the visual and the auditory modalities between them. TBI patients showed prolonged simple and choice RTs throughout all tasks, but their difficulty with integrating the tactile modality was disproportionately higher in comparison to controls. PMID- 12745714 TI - Assessing distress in caregivers of people with a traumatic brain injury (TBI): a psychometric study of the Head Injury Behaviour Scale. AB - This study examined the psychometric properties of the Head Injury Behaviour Scale (HIBS) using responses from 242 caregivers. The HIBS is a 20-item, scale describing common behavioural problems following traumatic brain injury (TBI), which are typical of dysexecutive syndrome. A principal components analysis was conducted on caregiver distress ratings for the 20 items comprising the HIBS and produced a two-factor solution with 10 items loading on each factor. Consequently, two sub-scales were derived and labelled Emotional Regulation and Behavioural Regulation. The sub-scales were found to have good internal consistency reliability. It is suggested that the Emotional Regulation sub-scale assesses behaviours reflecting impaired emotional control, occurring during interactions with caregivers. The Behavioural Regulation sub-scale assesses problems typical of dysexecutive syndrome, but which carry less emotional valence for caregivers. The clinical implications of these findings are discussed and normative data provided for the two sub-scales. PMID- 12745715 TI - Development and application of the Family Empowerment Questionnaire in brain injury. AB - BACKGROUND: The present study was an ongoing development of a previous study of the Family Empowerment Questionnaire, which had already indicated a four-factor solution (Skill, Knowledge, Support and Aspiration) empowerment process among family caretakers for their family members with traumatic brain injury (TBI). OBJECTIVE: The Family Empowerment Questionnaire was used to further explore its applicability to rehabilitation professionals through investigating differences between rehabilitation professionals' and family members' perceptions of empowerment in the rehabilitation process of persons with TBI. METHOD AND PROCEDURES: The present study compared the 52-item Family Empowerment Questionnaire scores between 65 rehabilitation professionals with the results obtained from 211 family caretakers. MAIN OUTCOME: Statistical analyses using the factor-based scores of the four sub-scales revealed that professionals and family members differ significantly in their perceptions regarding family empowerment in TBI rehabilitation in any of the four empowerment factors and within individual items of each factor. CONCLUSIONS: Specific activities in which families and professionals' perceptions differed significantly should be explored further and clinical implications of the study are discussed. PMID- 12745716 TI - Diminished expression of aquaporin water channels in ureteral-obstructed kidney in rats. AB - OBJECTIVE: We investigated whether regulation of aquaporin (AQP) water channels is altered in the ureteral-obstructed kidney. MATERIAL AND METHODS: Male Sprague Dawley rats were unilaterally obstructed of their left proximal ureters for 48 h. The protein expression of AQP1-3 channels was determined in the kidney by Western blot analysis. The expression of AQP2 was also determined by immunohistochemistry. In order to specifically determine primary impairment of the pathway leading to an altered regulation of AQP channels stimulated by the arginine vasopressin (AVP)/cyclic adenosine monophosphate (cAMP) pathway, the catalytic activity and protein expression of different parts of the adenylyl cyclase complex were separately determined. RESULTS: In the previously obstructed kidney, urinary osmolality and free water reabsorption were greatly decreased. The expression of AQP2 proteins was decreased in the cortex, outer medulla and inner medulla. Immunohistochemistry also showed a marked decrease in AQP2 expression. The expression of AQP1 and AQP3 was decreased in the outer medulla and inner medulla. cAMP generation in response to AVP, sodium fluoride or forskolin was greatly decreased. The expression of Gsalpha and adenylyl cyclase VI proteins was decreased. The contralateral kidney showed minimal or no changes in these parameters. CONCLUSIONS: The reduced abundance of AQP water channels may at least partly account for the urinary concentration defect in the ureteral obstructed kidney. The primary point of impairment of AQP channels regulated by the AVP/cAMP pathway may lie at the level of the catalytic unit of adenylyl cyclase. PMID- 12745717 TI - Bladder neoplasms--regions at chromosome 9 with putative tumour suppressor genes. AB - OBJECTIVES: To investigate the prevalence of loss of heterozygosity (LOH) at 12 different loci on chromosome 9 in patients with bladder neoplasms using a newly developed fluorescent multiplex polymerase chain reaction. PATIENTS AND METHODS: In a population-based study, freshly frozen tissue was collected from all cases of newly detected bladder neoplasms in the Stockholm region during 1995 and 1996 (n = 538) and 156 representative cases were subsequently studied in the present series. RESULTS: In total, at one or more loci of chromosome 9, 89% (139/156) of the tumours showed LOH. Loss of heterozygosity in informative cases was in the range from 33.1% (41/124) at the 9p21 locus to 67% (77/115) at the 9q31.3-32 loci. When minor LOH was studied, representing a single LOH with retention of heterozygosity at both adjacent markers, relatively frequent losses were detected at 9q22.3 harbouring the PTCH gene (7.7%), at 9q32-33.1 (6.6%) and at 9q33.2 harbouring the DBCCR1 gene (7.5%). In relation to clinical information, LOH at 9p22.1 was statistically significantly correlated with tumour grade (p = 0.01), but not with tumour stage. Replication errors were observed in 14 of 156 (9%) tumours. CONCLUSIONS: Our observation of relatively frequent minor LOH at 9p22.1, 9q22.3 and 9q32-33.1 identifies regions within which putative tumour suppressor genes, including the PTCH and the DBCCR1 genes, may reside. PMID- 12745718 TI - Validity of a population-based cancer register in Sweden--an assessment of data reproducibility in the South-East Region Prostate Cancer Register. AB - BACKGROUND: With a population-based setting, high coverage and accurately recorded data, the validity of a register is guaranteed. The South-East Region Prostate Cancer relies on the National Cancer Register as a basic source of data, thereby ensuring a high coverage of the corresponding geographic area. To assess the reproducibility of the data recorded a random sample of the cases were reviewed a second time and compared to the original recording. MATERIAL AND METHODS: The South-East Region Prostate Cancer Register was started in 1987. In addition to the basic data acquired from the Swedish National Register, it also includes tumour stage, grade, treatment and, since 1992, PSA. In the first stage of quality assessment 10 cases for each of the years 1987-1996 from Linkoping University Hospital were randomly selected for two independent recodings according to the same protocol as the original registration. In the second step 10 cases each for the same years from the remaining 8 hospitals in the region were selected for a single recoding. RESULTS: No systematic deviations were seen between the two independent recodings from Linkoping, a single recoding was therefore considered sufficient for assessing the reproducibility of the data from the remaining hospitals in the region. The Kappa values for agreement between the original registration and the single recoding ranged from 0.589 to 0.869. CONCLUSION: The population-based setting and high coverage guarantees the external validity of the register. The internal validity is ensured by the high reproducibility shown in the present study. PMID- 12745719 TI - Functional bladder capacity after bladder biofeedback predicts long-term outcome in children with nocturnal enuresis. AB - OBJECTIVE: We previously reported a 70% cure rate for bladder biofeedback in children with primary nocturnal enuresis associated with small bladder capacity and detrusor instability. In this paper we report on bladder capacity and incidence of enuresis after 60 months of follow-up and discuss the role of decreased bladder capacity in nocturnal enuresis. MATERIAL AND METHODS: We prospectively evaluated 21 boys and 3 girls (mean age 10.4 years) treated with bladder biofeedback between October 1993 and July 1995. Baseline bladder capacity and capacity at the end of treatment and at 60 months follow-up were determined from a micturition chart. RESULTS: At the end of primary treatment 17/24 patients had stopped bedwetting. In 4/17 responders and 4/7 non-responders the bladder capacity was <90% of normal for age. At 60 months, 4 patients had been lost to follow-up, 15 were dry at night and 4 continued bedwetting. One patient underwent surgery and was excluded from the study. Only 2/15 dry patients but 3/4 patients with persistent nocturnal enuresis had a bladder capacity of <90% of normal. CONCLUSIONS: Bladder biofeedback can be successfully used to treat children with refractory nocturnal enuresis associated with small bladder capacity and unstable detrusor. Normalization of bladder capacity and continuous growth of the bladder in order to keep the capacity normal would seem to be crucial to the long-term resolution of bedwetting in this select patient population. PMID- 12745720 TI - Self-construing in children with primary mono-symptomatic nocturnal enuresis--an investigation of three measures. AB - OBJECTIVE: This study aims to measure different aspects of self-construing in children with primary mono-symptomatic nocturnal enuresis. METHOD: 25 children aged 7-14 years, with nocturnal enuresis were recruited from a paediatric outpatient's unit specialising in enuresis [mean age 10.6 [males], 9.39 [females]] and 25 children acting as controls, were recruited from one primary and one secondary school [mean age 10.08 [males], 9.39 [females]]. MEASURES: The Butler Self Image Profile [SIP], Coopersmith Self Esteem Inventory, Ouvinen Birgerstam "I think I am" Self Perception Scale were administered. DESIGN: This study was a matched controlled group comparison. RESULTS: The only significant difference was the tendency for children with enuresis to construe themselves more negatively on the SIP. There were no significant differences between children with enuresis and the matched controls on all the other measures. CONCLUSION: Children with nocturnal enuresis generally perceive themselves similar to children without nocturnal enuresis, and are not "psychologically or emotionally disturbed." It is important to assess a child's self-construing in order to identify those children who are more vulnerable to a lower self-esteem. Future research needs to take into account self-construing in different clinical populations, i.e. those children who have non-mono symptomatic nocturnal enuresis. PMID- 12745721 TI - Pediatric urolithiasis--evaluation of risk factors in 95 children. AB - OBJECTIVE: Pediatric urolithiasis is a rarely encountered pathology, except in endemic areas such as Turkey. As a recurrent pathology which may reveal functional as well and morphologic changes in the urinary tract, metabolic and environmental factors, in addition to urogenital abnormalities, should be evaluated thoroughly in each patient. In this prospective study, the patient and family histories of 95 children with stone disease were evaluated, together with serum and urine risk factors. MATERIAL AND METHODS: Between 1996 and 2001, 95 children (25 females, 70 males; mean age 7.3 years; age range 0.6-15 years) referred to our department with urolithiasis were evaluated. All patients were investigated with respect to stone localization, associated abnormalities, urinary tract infection (UTI), positive family history and serum and urine risk factors. In addition to standard risk factors (hypocitraturia, hypercalciuria, hyperoxaluria, hyperuricosuria, hypomagnesuria), diet and 24-h urine volume were also assessed in all children. Children with cystinuria were excluded from the study. RESULTS: Stone size ranged from 0.3 to 3.3 cm, with an average value of 2.0 cm. The localization of the stones was classified as unilateral single stone in 37 patients, multiple unilateral stones in six and bilateral multiple stones in 27. Hypocitraturia was the commonest risk factor detected in our patients. A positive family history was present in 51 cases (54%). In addition, UTI was present in 59 cases (62%) and 67 cases had a previous history of recurrent UTI. Associated urogenital abnormality was detected in nine cases (9.4%). There were significant correlations between stone size and urinary citrate excretion (p < 0.05) and between the presence of UTI and urinary phosphate excretion (r = 0.59, p = 0.047). Treatments used were open surgery in seven (7.3%) cases, extracorporeal shock-wave lithotripsy in 39 (41%) and endoscopic surgery in 20 (21%). Following these procedures, 39 (41%) patients were completely stone-free, 11 (11%) had residual stones (<5 mm in diameter) and 12 (14.8%) passed the stone(s) spontaneously. During follow-up, regrowth was seen in four (4.2%) patients and stone recurrence was noted in a further four (4.2%). CONCLUSIONS: In addition to stone removal, treatment of pediatric urolithiasis requires a thorough metabolic and environmental evaluation of all patients on an individual basis. Obstructive pathologies have to be corrected immediately and apparent metabolic abnormalities should also be treated. Children with a positive family history should be followed carefully with respect to stone recurrence. Urine volume increases in parallel with body mass index and medical therapeutic agents which increase urine citrate levels should be encouraged. PMID- 12745722 TI - Outcome of new surgical techniques in hypospadia repair. AB - OBJECTIVE: Retrospectively we evaluated the outcome of all primary hypospadia operations done at Skejby Sygehus, Aarhus University Hospital, in the period May 1999 to March 2001. In that period the concept of treating hypospadias was changed from mainly the Mathiau operation and the preputial skin tube (Duckett) to the method of Barcat (and Koyanagi). MATERIAL AND METHODS: During the study period 101 boys consecutively underwent treatment of hypospadias using mainly the techniques of Koyanagi, Barcat and the Glanular Approximation Procedure (GAP). RESULTS: Cosmetic and functional outcome were excellent. Complications consisted of 12% urethrocutaneous fistula of which the majorities were seen in the first year after taken up the new techniques. There were 3% meatal stenosis and no urethral diverticula. CONCLUSION: The short-term results of the new techniques are superior in cosmetic and functional outcome. The fistula rate was a little high in the start of the period after introducing the new techniques but have fallen to a level comparable to other centres results. The level of fistula is comparable to other centres results. PMID- 12745723 TI - Retroperitoneal abscesses--analysis of a series of 66 cases. AB - OBJECTIVE: To analyze our experience with the management of retroperitoneal abscesses. PATIENTS AND METHODS: A retrospective study was made of 66 patients with retroperitoneal abscesses treated at our hospital from January 1975 to July 2001 for the purpose of analyzing the diagnosis and treatment of these rare infections. In each case, we analyzed patient characteristics, abscess location and origin, predisposing factors, clinical presentation, microbiology, radiographic findings, treatment, and outcome. RESULTS: In our series, the most frequent type of abscess was perinephric (45.4%), and the most frequent origin was the kidney (72.7%), generally renal lithiasis or previous urological surgery. Gram-negative bacilli were the microorganisms most often involved as causal agents of abscesses. CT had the best diagnostic performance (95%). Percutaneous drainage resolved the abscess in 86.3% of the patients in which it was used, compared with 87.5% for traditional surgical drainage. In 4 cases, the only treatment was administration of antibiotics. In all these cases the abscesses were smaller than 3 cm and patients were in good general condition. The mortality rate was excellent (1.5%), probably due to the low rate of comorbidity in our patients. CONCLUSIONS: Gram-negative bacilli were the most frequent microorganisms in our retroperitoneal abscesses. CT was the imaging technique that produced the most reliable and rapid diagnosis. Radiographically-guided percutaneous drainage was a safe and effective therapeutic alternative when used as definitive treatment or preoperatively. PMID- 12745724 TI - Prophylactic effects of systemic oral ephedrine in spinal anesthesia-induced hypotension during transurethral prostatectomy. AB - OBJECTIVE: We investigated the prophylactic effects of systemic oral ephedrine in spinal anesthesia-induced hypotension during transurethral prostatectomy. MATERIAL AND METHODS: Sixty American Society of Anesthesiologists Grade II and III patients scheduled for spinal anesthesia were randomized into one of two groups. Patients in Group I (n = 30) received oral ephedrine 50 mg in addition to premedication whilst those in Group II (n = 30) received only premedication 30 min before spinal anesthesia. Pre-infusion values were measured in order to obtain baseline readings after oral ephedrine administration in Group I and after premedication in Group II. Systolic arterial pressure (SAP) and heart rate (HR) were recorded before and after infusion, during and 5 min after spinal anesthesia and intraoperatively. Hypotension was defined as SAP <100 mmHg and <20% of baseline value. Hypotension was treated with 3 mg ephedrine and bradycardia was corrected with atropine 0.5 mg, given as an i.v. bolus. RESULTS: SAP values were significantly lower in Group II during the spinal anesthesia, post-spinal and intraoperative periods (p < 0.0001). Fifteen patients received ephedrine in Group II and seven in Group I. Supplemental ephedrine was used at doses of 3.42 +/- 0.97 mg in Group I and 8.86 +/- 1.24 mg in Group II. The incidence of hypotension was halved in Group I compared to Group II (23.33% vs 50%, p = 0.003). Six patients received atropine in Group II because of severe bradycardia. Mean HR values were lower in Group II than Group I during the spinal anesthesia, post spinal and intraoperative periods. CONCLUSIONS: We conclude that a prophylactic oral dose of ephedrine 50 mg is effective for minimizing and managing spinal anesthesia-induced hypotension during transurethral prostatectomy. PMID- 12745725 TI - Microalbuminuria in diabetic and hypertensive patients and the general population -consequences of various diagnostic criteria--the Nord-Trondelag Health Study (HUNT). AB - OBJECTIVE: The purpose of this paper was to study the prevalence of microalbuminuria (MA) in males and females of various ages by applying various diagnostic criteria. Three groups of subjects were studied: apparently healthy individuals; self-reported hypertensives; and diabetics. MATERIAL AND METHODS: A total of 9255 individuals (age > or =20 years), all of whom were identified from the large (n = 65 258) population-based Nord-Trondelag Health Study (HUNT) performed in Norway between 1995 and 1997, delivered three morning urine samples for MA analysis. Of these individuals, 651 reported both diabetes and treated hypertension, 944 diabetes only and 5547 treated hypertension only. The remaining 2113 subjects without diabetes or treated hypertension were randomly selected. The albumin:creatinine ratio (ACR) was used as an expression of urine albumin excretion. RESULTS: Applying the classical definition of MA of ACR > or =2.5 mg/mmol in at least two out of three urine samples, the prevalence of MA in those with both diabetes and hypertension was 42.2% in males and 25.9% in females; corresponding values for those with diabetes only were 27.8% and 22.4%, for the hypertensives 19.3% and 11.5% and for the randomly selected sample 5.2% and 4.7%. The prevalence of MA increased strongly with increasing age for both genders in all subgroups. The prevalence of MA changed considerably when applying different cut-off values of ACR and at least one, two or three urine samples with ACRs above the cut-off value. CONCLUSIONS: This study, one of the largest cross sectional screening studies of MA ever performed, clearly illustrates the consequences of applying different diagnostic criteria. The optimal cut-off levels of MA for the prediction of cardiovascular disease still remain to be properly defined, and more follow-up studies are therefore needed. PMID- 12745726 TI - Functional evaluation of the remaining kidney in patients after unilateral nephrectomy. AB - BACKGROUND: Unilateral nephrectomy is quite often surgical procedure. The remaining kidney undergoes a sequel of adaptational processes. The aim of the study was to evaluate kidney function in patients subjected to unilateral nephrectomy. MATERIALS AND METHODS: The study was carried out in 28 subjects allocated into three groups: healthy controls (n = 8) and patients subjected to unilateral nephrectomy evaluated 1 month (n = 10) and 1 year (n = 10) from the surgery. Biochemical as well ultrasonographic and scintigraphic data were recorded. RESULTS: From all evaluated standard biochemical parameters (creatinine, creatinine clearance, urea, microalbuminuria) significant changes were observed in the case of creatinine and microalbuminuria levels at 1 month, which increased from 0.96 mg/ml to 1.05 mg/dl and from 5.14 mg/24 h to 20.0 mg/24 h, respectively. (99)Tc(m)-DTPA plasma clearance was significantly elevated in patients 1 month after unilateral nephrectomy, by 7.5%, with a decrease by 17% in patients 1 year after surgical procedure, in reference to the control subjects. A significant increase in (99)Tc(m)-EC plasma clearance of patients evaluated 1 year from the operation, by 13% (p < 0.05) in comparison to the control group was seen. RI index markedly increased in nephrectomised patients both after 1 month and 1 year from the operation as compared to the controls, from 0.59 to 0.64 (p < 0.05) and 0.63 (p < 0.05), respectively. CONCLUSION: Adaptational changes of the remaining kidney are observed in patients 1 month and 1 year after unilateral nephrectomy. PMID- 12745727 TI - Interleukin-17 stimulates the release of pro-inflammatory cytokines by blood monocytes in patients with IgA nephropathy. AB - OBJECTIVE: Interleukin-17 (IL-17) is a newly discovered cytokine implicated in the regulation of inflammation. The present study was designed to explore whether IL-17 is involved in the immunoregulatory response in IgA nephropathy (IgAN). MATERIAL AND METHODS: We examined the in vitro effect of recombinant human IL-17 (rhIL-17) on pro-inflammatory cytokine release by peripheral blood monocytes (PBM) from patients with IgAN. Measurement of IL-1beta and tumor necrosis factor alpha (TNF-alpha) was performed by means of a sandwich enzyme-linked immunosorbent assay. RESULTS: IL-1beta and TNF-alpha release were upregulated by rhIL-17 in a dose- and time-dependent fashion. Treatment of PBM from patients with IgAN with lipopolysaccharide and rhIL-17 resulted in significant activation and release of IL-1beta and TNF-alpha protein. Levels of IL-1beta and TNF-alpha were significantly higher in IgAN patients with the nephrotic syndrome (NS) than in patients without NS or in healthy subjects. We also provide information indicating that there is excessive production of IL-17 in IgAN patients. When IL 17-activated PBM were incubated in the presence of IL-10, IL-10 exerted a significant suppressive effect on IL-1beta and TNF-alpha release in vitro. CONCLUSION: IL-17 can stimulate the release of pro-inflammatory cytokines from PBM in patients with IgAN. PMID- 12745728 TI - Maintaining patency of tunneled hemodialysis catheters--efficacy of aspirin compared to warfarin. AB - OBJECTIVE: Tunneled hemodialysis catheters (caths) often fail as a result of luminal obstructive thrombus or formation of a fibrin sheath at the tip. Anecdotal and non-randomized studies have indicated that aspirin (A) and/or warfarin (W) can prolong cath patency. We examined the effect of chronic usage of either A or W on primary cath patency. MATERIAL AND METHODS: A prospective cross sectional monitoring of cath patency was conducted over a 3-year period. Patients were grouped according to their long-term usage of either A (325 mg daily) or W. Patients on neither medication served as a control (C). The end point of the study occurred at cannulation of the patients' arteriovenous fistulae, when there was development of cath-related bacteremia or when there was inability to maintain a blood flow of 250 ml/min. RESULTS: Sixty-three patients with a mean age of 57 +/- 15 years completed the study. There were 21 patients in the A group, 11 in the W group and 31 in the C group. Cath survival was 91%, 73% and 29% at 120 days for the A, W and C groups, respectively (A vs C, p < 0.0001; W vs C, p < 0.0001; A vs W, p = NS). The mean durations of cath patency were 114 +/- 18, 111 +/- 17 and 68 +/- 37 days for the A, W and C groups, respectively (A vs C, p < 0.0001; W vs C, p < 0.0001; A vs W, p = NS). Gastrointestinal (GI) bleeding complication rates were 24%, 18% and 0% for the A, W and C groups, respectively (A vs C, p = 0.02; W vs C, p = 0.02; A vs W, p = NS). The relative risk of GI bleeding associated with aspirin was 0.71 [95% confidence interval (CI) 0.11-4.4, p = 0.7] but among elderly aspirin users it was 1.14 (CI 1.0-1.3, p = 0.008). CONCLUSION: Both aspirin and warfarin are equally effective at prolonging cath patency but their routine use for failing caths cannot be unequivocally recommended because of the increased risk of GI bleeding. Further prospective and randomized studies are called for. PMID- 12745729 TI - Malignant transformation in a mature testicular teratoma left untreated for more than 50 years since childhood. AB - Although testicular teratoma in childhood is regarded as a benign tumor, little is known about the consequences of pediatric teratoma being left untreated. We report herein a case of malignant transformation observed in a mature testicular teratoma that was presumed to have remained benign for >50 years. PMID- 12745730 TI - Paratesticular adenofibroma. AB - Paratesticular adenofibroma is an uncommon benign tumour. Although it has previously been reported in the female reproductive organs, to our knowledge only three cases have previously been reported in the male genital organs. Herein we describe an adenofibroma that developed in the tunica vaginalis and rete testis. We discuss its histogenesis and conservative treatment. PMID- 12745731 TI - Seminoma in an intra-abdominal testis misdiagnosed as a kidney tumor and associated with ipsilateral renal agenesis. AB - We present a case of seminoma found in an intra-abdominal testis and associated with ipsilateral renal agenesis and absence of the ipsilateral seminal vesicle. Although malignant tumor degeneration in an undescended intra-abdominal testis is rare, understanding its radiologic appearance is important in order to confirm the diagnosis preoperatively and detect possible malformations of other organs. PMID- 12745732 TI - Urethral erosion of tension-free vaginal tape. AB - We report a case of urinary retention and urethral erosion after placement of tension-free vaginal tape. This procedure is gaining wide popularity in both Europe and the United States. Careful attention is necessary to assure tension free placement of the sling. PMID- 12745733 TI - Successful conservative management of a pyelo-duodenal fistula. AB - Uro-intestinal fistula is a rare entity and mostly managed by means of surgical intervention. We report a case of pyelo-duodenal fistula which was treated conservatively. PMID- 12745734 TI - Multiple myeloma presenting as bilateral renal complex stones. AB - Hypercalcaemia is a frequent complication of multiple myeloma. A mild degree of nephrocalcinosis has been noted in occasional patients and renal calculi are also sometimes present. We report herein a case of multiple myeloma that was revealed by the presence of bilateral calcium renal staghorn calculi. Medical treatment of myeloma, surgical management of this complex lithiasis and the course of multiple myeloma are discussed. PMID- 12745735 TI - Psychological aspects of screening in families with hereditary prostate cancer. AB - OBJECTIVE: Approximately 5-10% of prostate cancer cases are caused by dominantly inherited susceptibility to the disease. Although advances have been made in research concerning the genetic mechanisms of hereditary prostate cancer, little is known about the psychological consequences. The aim of this study was to assess possible negative psychological effects of screening for prostate cancer in a high-risk population. MATERIAL AND METHODS: This study was based on a previous study of risk perception, screening practice and interest in genetic testing among unaffected men in families with hereditary prostate cancer. The present study included 87 men from the previous study who were screened regularly for prostate cancer. Of these, 74 men agreed to receive two further questionnaires, both of which included the Hospital Anxiety and Depression Scale (HAD) and the Impact of Event Scale (IES), one of which was filled in on the day of the next screening visit and the other 4-6 weeks later. RESULTS: The response rate was 77% (57/74). There were no statistically significant differences in total or subscale HAD or IES scores between the two points of measurement. There was a trend towards slightly higher HAD scores on the day of the screening visit, but the difference was so small that we did not consider it clinically relevant. In an attempt to identify risk factors for a negative impact of screening several subgroup analyses were performed, but none of these subgroups had significantly higher scores on the day of the visit than afterwards. CONCLUSION: Most men with a high hereditary risk of developing prostate cancer do not experience severe psychological adverse effects resulting from attendance for screening. PMID- 12745737 TI - Magnetic resonance urography in pediatric urology. AB - OBJECTIVE: To evaluate the efficiency of magnetic resonance urography (MRU) in pediatric urology. MATERIAL AND METHODS: We report retrospectively on 12 children who underwent MRU between January 1999 and November 2001. MRU was performed to accurately evaluate the entire urinary tract because of megaureter, ectopic ureter, vesicoureteral reflux, Y-inverted duplication and hydronephrosis because of pyeloureteral stenosis. T1- and T2-weighted images were obtained in the coronal, sagittal and axial planes. The mean age of the children (8 females, 4 males) investigated was 36 months (range 2-140 months). RESULTS: An accurate anatomical picture of the entire urinary tract could be obtained in all children. The obstructive nature of megaureter could be differentiated. The distal orifice of ectopic ureter could be identified in the vagina. Vesicoureteral reflux into the blind-ending ureteral bud of a duplicated system was accurately identified. Hydronephrosis was demonstrated to be the result of pyeloureteral stenosis. The location of stenoses was easily identified in the sagittal and coronal planes. CONCLUSIONS: MRU is an excellent imaging modality for accurately depicting the urinary tract. MRU is superior to conventional intravenous urography because it does not use ionizing radiation, the gadolinium contrast medium used is not nephrotoxic and the imaging quality is excellent, reproducible and not interfered with by gas superposition. Considering the high costs and diagnostic benefit of MRU compared to intravenous urography, MRU should be performed in patients with impaired renal function, in those with an allergy to contrast medium and if anatomic relationships are not clear prior to reconstructive surgery. PMID- 12745736 TI - Postoperative radiotherapy after prostatectomy--a review. AB - PURPOSE: The management of prostate adenocarcinomas using postoperative irradiation is a controversial question. The purpose of this study was to review the literature on the subject. MATERIAL AND METHODS: A total of 417 articles dealing with postoperative radiotherapy after radical prostatectomy in English literature (1990-2002) were reviewed in aspects of effect on survival, time of irradiation, risk factors, dose and technique and side effects. RESULTS AND DISCUSSION: No randomised studies have been performed and therefore no definitive conclusive data can be made concerning the efficiency of the concept. However, postoperative radiotherapy appears to increase local control preferably in pT3/4 prostatic carcinomas with seminal vesicles involvement and/or positive margins and/or high Gleason score and high postoperative PSA level. It has not been shown to improve survival. Severe side effects are reported in a low frequency. However, postoperative irradiation can cause severe side effects and postoperative adjuvant/salvage treatments should be delivered earliest 3-6 months after surgery and the total dose delivered to the prostate bed should be 65-70 Gy. Postoperative radiotherapy induces improved local control in patients with positive surgical margins and in patients with a local relapse, preferably if the tumour is small (i.e. PSA <1-2 ng/mL). PMID- 12745738 TI - The Canadian Enuresis Study and Evaluation--short- and long-term safety and efficacy of an oral desmopressin preparation. AB - OBJECTIVE: To evaluate the long-term (12 months) efficacy and safety of oral desmopressin (DDAVP). MATERIAL AND METHODS: A total of 256 healthy children (6-18 years old) with nocturnal enuresis with a frequency of > or = 10 wet nights during a 4-week observation period were eligible for inclusion in the study. Initially 0.2 mg of DDAVP was given for 14 nights. Those achieving a > 90% reduction in the number of wet nights over the observation period (full responders) began a 12-week continuous treatment period at this dose. The remaining children received 0.4 mg for an additional 14 nights. Those achieving a > or = 50% reduction in the number of wet nights (responders) commenced a 12-week continuous treatment period at this dose. Children with a < 50% reduction in the number of wet nights at this point were withdrawn from the study. Each 12-week treatment period was followed by a treatment-free period of 7-28 days. Children who remained dry during that period were assigned a full response and terminated the trial. Children with > or = 2 wet nights during that period immediately began a new 12-week treatment period at the previous dose. This was repeated for 12 months and thereafter the medication dose was tapered by halving over a 4-week period. RESULTS: A total of 117/236 children who completed the titration period (49.6%; 95% confidence interval 40-57%) responded (> 50% reduction over baseline). Throughout the study their response rate remained constant at approximately 74%. Continuous treatment reduced the median number of wet nights during the observation period from 5.75 to 1.00 per week. A total of 12.4% of children received the 0.2 mg dose and 87.6% the 0.4 mg dose. The proportion of full responses increased over the course of the study from 5.8% to 37.5%. DDAVP was well tolerated: the majority of reported adverse events were mild, although two adverse events leading to withdrawal were reported. CONCLUSIONS: Oral DDAVP provides an effective and well-tolerated means of providing long-term control in children with nocturnal enuresis. Long-term treatment increases the response rate. PMID- 12745739 TI - Renal pelvic wall thickening in childhood urinary tract infections--evidence of acute pyelitis or vesicoureteral reflux? AB - OBJECTIVE: The aim of this study was to determine whether renal pelvic wall thickening in active childhood urinary tract infections (UTIs), as demonstrated using ultrasound, is caused by acute pyelitis or by vesicoureteral reflux (VUR) related chronic changes. MATERIAL AND METHODS: A total of 41 children with at least unilateral renal pelvic wall thickening as demonstrated using ultrasound during the acute stage of UTI, and confirmed using voiding cystourethrography (VCUG), were analyzed. All cases underwent ultrasound studies at 6 months follow up. RESULTS: Using ultrasound, 50 halves of the pelvis showed renal pelvic wall thickening. In 27 (54%), ipsilateral VUR could be demonstrated using VCUG. The sensitivity, specificity and positive predictive value of renal pelvic wall thickening for predicting ipsilateral VUR were 79.4%, 52.1% and 54%, respectively. Only 2 (7%) cases presented with wall thickening at 6 months follow up. Most of the thickening recovered after clinical improvement, although VUR became persistent in half the cases. CONCLUSIONS: Renal pelvic wall thickening is an abnormal finding in the acute stage of childhood UTI and predominantly indicates acute pyelitis rather than VUR-related chronic changes. Renal pelvic wall thickening is not sufficiently predictive of VUR in acute UTI, although it does provide evidence of upper UTI. PMID- 12745740 TI - TUMT 2.0: results three months and three years after treatment. AB - OBJECTIVE: To examine the durability of the results following low-power transurethral microwave thermotherapy (TUMT). MATERIAL AND METHODS: 28 patients 55 to 83 years of age with lower urinary tract symptoms (LUTS) and marginal/moderate infravesical obstruction and 4 patients with LUTS, peak urinary flow (Qmax) less than than 15 ml/sec and prostate volume below 40 ml. We used a Prostatron version 2.0. RESULTS: transurethral resection of the prostate (TURP) was done shortly after treatment in one patient with acute obstruction caused by necrotic tissue. Three months after treatment 15 patients reported that they were cured from LUTS and 10 experienced improvement of symptoms. A significant reduction of the symptom scores was seen among the 31 patients not operated while Qmax was unchanged. Three years after treatment two patients were still cured from LUTS, 9 hadsome reduction of symptoms, and TURP had been done in further three patients. Symptom scores among the 24 patients still in the study were significantly lower than the baseline values but also significantly higher than the symptom scores three months after treatment. Residual urine was reduced while no changes were seen in Qmax. CONCLUSION: TUMT done by Prostatron version 2.0 causes mainly a reduction of symptoms. Most of the effect disappears after a few years. PMID- 12745741 TI - Variability of the International Prostate Symptom Score in men with lower urinary tract symptoms. AB - OBJECTIVE: The variability of the International Prostate Symptom Score (I-PSS) was tested in patients with benign prostatic hyperplasia (BPH) by comparing questionnaire results obtained in the physician's office and, 1 week later, in the patient's home. MATERIAL AND METHODS: A total of 210 consecutive men with lower urinary tract symptoms (LUTS) completed the I-PSS questionnaire in the physician's office. One week later the questionnaire was mailed to each patient's home, completed by the patient and then returned. Scores were compared specifically in terms of clinically significant differences defined by a total symptom score difference of > or = 6 points and/or a difference of > or = 2 points in the quality-of-life (QOL) measure. RESULTS: The mean patient age was 67 years. Questionnaires were completed and returned by 127/210 (60%) men. Pearson's correlation coefficient for the I-PSS and QOL results was 0.81 and 0.74, respectively. Clinically significant differences in results were seen in 33/127 (26%) patients. When grouping patients into mild, moderate and severe symptom categories based on the I-PSS results, 31/127 (24%) changed categories when comparing "office" and "home" results. CONCLUSIONS: Clinically significant variations in I-PSS and QOL results may exist and may affect treatment decisions in > 25% of men. PMID- 12745742 TI - Effect of varicocele on testicular artery blood flow in men--color Doppler investigation. AB - OBJECTIVE: Varicocele can be defined as an abnormal tortuosity and dilatation of the veins of the pampiniform plexus. Contradictory results have been obtained from experimental animal models and a few clinical human studies on testicular arterial blood flow in varicocele. The purpose of this study was to determine the changes in testicular arterial blood flow parameters in patients with varicocele. MATERIAL AND METHODS: A total of 62 patients with a clinical diagnosis of left varicocele and a scrotal vein with a diameter of > or = 3 mm on color Doppler ultrasonography were included in the study. A total of 44 fertile normal male volunteers served as controls. RESULTS: Median testicular arterial blood flow and median flow rate in milliliters per minute per 100 g of testicular tissue were found to be significantly decreased in the patient group compared to the control group: blood flow, 1.42 and 2.00 ml/min; flow rate, 9.63 and 12.35 ml/min/100 g, respectively (p < 0.05). Positive correlations were found between sperm concentration and left testicular artery blood flow (p < 0.05) and between left testicular volume and testicular artery blood flow (p < 0.05). CONCLUSIONS: Testicular arterial blood flow was found to be significantly decreased in men with varicocele. This may be a reflection of the impaired microcirculation. Following decreased testicular arterial blood flow, impaired spermatogenesis may result from defective energy metabolism in the microcirculatory bed. PMID- 12745743 TI - A new clinical classification for undescended testis. AB - A new classification for undescended testis (UDT), suitable for a clinical setting, is proposed. UDT is categorized into congenital and acquired forms. Congenital forms include intra-abdominal, intra-canalicular, supra-scrotal and ectopic testes. Acquired forms can be divided into primary and secondary types. Primary forms are described as either ascending testes, i.e. those which cannot be manipulated into the scrotum, or high scrotal testes, i.e. those which can still be brought through the scrotal entrance into a high scrotal (unstable) position. Secondary forms are the result of ipsilateral groin surgery and are termed "trapped testes". Congenital forms of UDT should be treated surgically at an early age, preferably at 1 year. Therapy for primary acquired forms remains controversial. Therapeutic modalities include orchidopexy, hormonal treatment (preferably with human chorionic gonadotrophin) or waiting for spontaneous descent during the peripubertal period ("laissez faire policy"). Secondary acquired forms are probably best treated surgically. PMID- 12745745 TI - Subinguinal microsurgical ligation--its effectiveness in pediatric and adolescent varicocele. AB - OBJECTIVE: Treatment of varicocele using a microsurgical vein ligation approach is a successful procedure when applied to adult patients. We performed this procedure in prepubertal and pubertal patients in order to verify its feasibility and effectiveness in a younger population. MATERIAL AND METHODS: Between January 1998 and December 1999, 46 patients (average age 13 years) were operated on for a left varicocele using a microsurgical selective vein ligation procedure. All patients were evaluated clinically and using color Doppler ultrasound (CDUS): 34/46 patients were classified clinically as grade III and the remaining 12 as grade II. We used a magnification loupe (x3) in 34/46 patients and an operating microscope (x 10-16) in the remaining 12. RESULTS: The procedure was successful in all but one patient (2.1%) who showed recurrence of the disease at the time of the first postoperative follow-up CDUS scan and required a repeat procedure. The average operative times were 45 and 60 min for loupe and microscopic procedures, respectively. In three cases (6.5%), early postoperative complications (wound infection and transient hydrocele) occurred and quickly resolved. All the patients were discharged within 24 h, returned to normal daily life within 2 days and resumed physical and sporting activities within 1 week. CONCLUSION: Subinguinal "artery- and lymphatic-sparing" microsurgical varicocelectomy, considered the gold standard treatment in adults, also proved to be successful in this preliminary experience with pediatric patients. The minimally invasive approach ensures the prevention of recurrences and/or postoperative complications, and is associated with a very low morbidity rate. PMID- 12745744 TI - Proliferating cell nuclear antigen in testes of infertile men with varicocele- preliminary results of interrelationship with sperm count before and after varicocelectomy. AB - OBJECTIVE: This study was conducted to assess germ cell kinetics and correlate them with sperm counts before and after varicocelectomy. MATERIAL AND METHODS: Forty-three testicular biopsy specimens were obtained from 47 patients with varicocele during varicocelectomy. Similar specimens were obtained from 8 fertile volunteers. All specimens were immunostained using anti-proliferating cell nuclear antigen (anti-PCNA) antibody. PCNA expression was evaluated by assessing its staining intensity (SI) and labeling index (LI). RESULTS: The varicocele specimens revealed significantly lower SI and LI than the controls. There was a significant correlation between initial sperm concentration and LI but not SI. Coexistence of LI > or = 31 and preoperative sperm count > 1 million/ml correlated with a significant rise in the postoperative sperm count. CONCLUSIONS: PCNA is a useful molecular marker for assessing germ cell kinetics in varicocele patients. The decline in DNA synthesis, as suggested by the lower PCNA SI and LI in varicocele specimens, could be a reason for the disordered spermatogenesis in these patients. PCNA LI may also be considered as a beneficial clinical marker and may help to predict the surgical outcome after varicocele repair. PMID- 12745746 TI - Nitric oxide and endothelin concentrations during intravenous infusion of urological irrigating fluid. AB - OBJECTIVE: To study the possible role of two vasoactive mediators, nitric oxide (NO) and endothelin, in the hemodynamic alterations following absorption of irrigating fluid. MATERIAL AND METHODS: Twelve male volunteers received an intravenous administration of 1.5% glycine, given at a rate of 0.5 ml/kg/min for 30 min. Infusions of 3% mannitol served as controls. Blood pressure, heart rate and the plasma levels of endothelin and nitrite/nitrate (an index of NO activity) were measured before, during and after the infusions. RESULTS: The endothelin concentration at baseline correlated inversely with the systolic arterial pressure (p<0.01). No qualitative differences were found between the two fluids with respect to the NO and endothelin responses, but the observed variations indicated the role of the two fluids during volume loading. In general, an elevation of blood pressure during the infusions was associated with an increase in the endothelin concentration, while a sustained elevation 30 min later was accompanied by a decrease in the NO activity. Reductions in the blood pressure and heart rate were associated with decreased NO concentrations. CONCLUSION: The endothelin/NO axis may increase blood pressure in response to volume loading; it also acts to stabilize the circulation at baseline and probably also when the blood pressure decreases. PMID- 12745747 TI - Intercurrent autoimmune conditions in classic and non-ulcer interstitial cystitis. AB - OBJECTIVE: Interstitial cystitis (IC) is known to be one of the most bothersome conditions encountered in urological practice. It is frequently divided into two subtypes: classic and non-ulcer. Several authors have reported on autoantibodies in patients with IC and clinical and histopathological findings show similarities with those in some autoimmune disorders. Furthermore, IC has been shown to be associated with autoimmune diseases such as systemic lupus erythematosus, Sjogren's syndrome and autoimmune diseases of the thyroid gland. Our aim was to study the occurrence of associated autoimmune conditions in patients with IC diagnosed according to the National Institutes of Health/National Institute of Arthritis, Diabetes, Digestive and Kidney Diseases criteria. MATERIAL AND METHODS: Our sample of IC patients consisted of 129 with classic IC and 93 with non-ulcer IC. Clinical records, including micturition charts, were surveyed, and a more in-depth interview was obtained by means of a telephone call. Autoimmune disorders, diagnosed by Swedish clinicians according to acknowledged diagnostic criteria, were registered. RESULTS: Allergy was the most common IC-associated condition, 41% of all patients with classic IC and 47% of all patients with non ulcer IC having some type or degree of hypersensitivity/allergy. Rheumatoid arthritis occurred in 13% of patients with classic IC and in 4% with non-ulcer IC. Inflammatory bowel disease was not diagnosed in any of the patients with non ulcer IC whereas 2.3% of the patients with classic IC had either ulcerative colitis or Crohn's disease, approximately 33 times the prevalence seen in the general population. CONCLUSION: It appears that systemic and autoimmune disorders are more prevalent in the IC population than in the general population. PMID- 12745748 TI - Urinary stone disease in diabetes mellitus. AB - OBJECTIVE: Diabetes mellitus (DM) and urinary stone disease (USD) are common diseases in the community. The prevalence of USD has not previously been studied in DM patients. MATERIAL AND METHODS: We evaluated 286 diabetics and 111 age matched controls by means of direct urinary system X-ray, ultrasonography and/or intravenous pyelography. We also examined the history, risk factors and clinical aspects of USD in DM patients. RESULTS: The prevalence of USD (21% vs 8%; p < 0.05) and the rate of recurrence (2.1 +/- 2.2 vs 1.3 +/- 0.5 stones/case; p < 0.05) were higher in the diabetics compared to the controls. Family history and male gender were significant risk factors for the development of USD in the DM patients. Increased daily total fluid consumption was not preventive for USD in the DM patients, and the type of fluid consumed did not have an impact on the risk of USD. Alcohol consumption was a significant risk factor for the development USD in the DM patients (odds ratio 3.68; 95% confidence interval 1.29 10.45; p < 0.05). Crystaluria (10% vs 1%; p < 0.05) and positive urine culture (8% vs 1%; p < 0.05) were also more prominent in the diabetics compared to the controls. CONCLUSIONS: The results of this study suggest that DM is a risk factor for the development USD. PMID- 12745749 TI - The comparison of antibody response to influenza vaccination in continuous ambulatory peritoneal dialysis, hemodialysis and renal transplantation patients. AB - BACKGROUND: The immune system in renal transplant (Tx), Continuous Ambulatory Peritoneal Dialysis (CAPD) and hemodialysis (HD) patients have been suppressed and antibody response to vaccination is weaker than that of the normal population. Additionally immune response to vaccination also differs from each other in aforementioned three groups resulting from different levels immunosuppression. In the present study, detection of antibody response to influenza vaccine as an indicator of the level of immunity in Tx, CAPD and HD patients was aimed PATIENTS AND METHODS: Forty-eight patients (17 Tx, 16 CAPD and 15 HD) and 10 healthy adults, as a control group were enrolled into the study. Purified, split-virus, commercial trivalent influenza vaccine (VAXIGRIP--Pasteur Merieux Connaught, single dose of 0.5 ml into the deltoid muscle) containing 15 microg of each hemagglutinin of A/Johannesburg/82/96 (H1N1), A/Nachang/933/95 (H3N2) and B/Harbin/07/94 (B) strains were administered to all subjects. Serum samples were collected before and 1 month after vaccination to determine antibody titers. Hemagglutination-inhibition test (HI) was applied for determination of antibody response. The antibody response against each strain was measured separately. In addition to measurement of antibody response, increments in antibody titer (n-fold increase in titer), proportion of patients with protective antibody levels and seroconversion levels were taken into account. Wilcoxon paired 2 test and Mann-Whitney U test were applied for statistical analysis. p < 0.05 was accepted as significance level. RESULTS: Significant increases in antibody titers for all three antigens were observed in the study groups after vaccination (p = 0.001). However, the increase in titer of H3N2 was lower in Tx, CAPD and HD patients than that of the control group (1.0-2.0 vs 5.00) (p = 0.01). The proportion of protective antibody titers and seroconvertions were increased after vaccination in all subjects. Proportions of patients with protective antibody titers after vaccination were lower in Tx, CAPD and HD groups in comparison to control group. CONCLUSION: Although antibody titers in Tx, CAPD and HD patients presented significant increases after vaccination, the proportions of patients with protective antibody titers were lower in comparison to control group. Tx, CAPD and HD patients should be vaccinated every year to be able avoid potential morbidity and mortality of the influenza infection. Trial of high dose vaccination protocols may be useful to increase the proportion of patients with protective antibody levels. PMID- 12745750 TI - The effects of intravenous iron treatment on oxidant stress and erythrocyte deformability in hemodialysis patients. AB - BACKGROUND: It is well known that free iron causes oxidant stress to increase. However data concerning whether intravenously (I.V) administered iron in maintenance doses (10-20 mg) gives rise to increased oxidant stress and disturbed erythrocyte deformability (EDEF) in hemodialysis (HD) patients is lacking. In the present study, we aimed to evaluate and compare the effects of I.V iron on oxidant stress and EDEF. PATIENTS AND METHODS: Thirteen HD patients (10 males, 3 females, mean age: 49.9 +/- 13.4 years), given I.V iron were included in the study. All patients were undergone three consecutive HD session. The first HD session was performed without iron administration (Group 1), whereas in the following sessions the same patients were given 20 mg (Group 2) and 100 mg (Group 3) iron III hydroxide sucrose (Venofer--Abdi Ibrahim) I.V at the end of the dialysis session. In study periods, 7 blood samples were drawn from each patient: before dialysis, at the end of the dialysis (just after the session), 15, 30, 60, 90 and 120 minutes after each dialysis session. However 15 minute samples were not drawn in the third group, since I.V iron was given by infusion in 30 minutes. EDEF and plasma malondialdehyde (MDA) were studied in all samples. RESULTS: When the results of the session without iron were considered, bivariate correlation analysis did not reveal any correlation between MDA and EDEF. When the course of each parameter were considered separately, MDA levels 90 and 120 minutes after HD session were significantly higher than that of the before and just after the HD session (p < 0.05). Whereas EDEF in 60, 90 and 120 minutes after HD session was found to be worsened when compared to before and just after HD sessions' values (p < 0.05). When results of the session with 20 mg iron were considered, EDEF and MDA values were not found to be correlated and throughout the course. Although EDEF did not present any significant change, MDA levels 60, 90 and 120 minutes after HD session were found to be significantly higher than that of the 15 and 30 minutes after HD session (p < 0,05). When results of the session with 100 mg iron were considered, MDA levels 30, 60, 90 and 120 minutes after HD session were found to be significantly higher than that of the before and just after the HD sessions' (p < 0,05). EDEF in 90 and 120 minutes after HD session was improved and no correlation between MDA and EDEF was observed. When groups were compared with each other, plasma MDA levels in session with 100 mg iron at the beginning, at the end and 30 minutes after HD were significantly lower than that of the without iron group (p < 0.05). Similarly MDA levels in session with 100 mg iron at the beginning, at the end, 30 minutes and 120 minutes after HD were significantly lower than that of the 20 mg iron (p < 0.05). When EDEF values in sessions with 20 mg iron and without iron were considered, only values 60 and 90 minutes after dialysis were significantly improved in 20 mg iron group. The others were statistically similar. CONCLUSION: In the present study, it was observed that I.V administered iron in 20 and 100 mg doses did not cause additional deteriorating effect on oxidant stress and EDEF was even improved by I.V iron. PMID- 12745751 TI - Idiopathic priapism in a young boy. AB - We report a case of recurrent priapism in a boy. There were no recurrences during or after oral treatment with Bricanyl (terbutaline). PMID- 12745752 TI - Pseudosarcomatous fibromyxoid tumor of the prostate. AB - A very rare case of benign infravesical obstruction due to a pseudosarcomatous fibromyxoid tumor of the prostate is presented. This entity is clinically and histologically difficult to distinguish from a malignant lesion. A discussion of the histopathological features and a review of the literature is given. PMID- 12745753 TI - Ureteral wound caused by blunt abdominal trauma. AB - A man fell on icy ground whilst walking to an outdoor toilet. An initial CT scan with intravenous contrast medium was negative. As the man experienced increasing pain a plain abdominal radiograph was performed 2 h later and revealed extravasation of contrast medium emanating from a ureteral injury. This case underlines the possibility that important injuries may not be visible on the initial CT scan that is often used in trauma diagnostics. PMID- 12745754 TI - Wunderlich syndrome as first manifestation of infective endocarditis. AB - Infective endocarditis (IE) presents with several signs and symptoms that are mainly heart-related and the result of bacteremia. We describe the case of a woman with severe renal hemorrhage due to a septic embolic cortical infarction, who was also receiving anticoagulation therapy because of cardiopathy, whose retroperitoneal hematoma was the first manifestation of IE. PMID- 12745755 TI - Renal hypomagnesemia, hypercalciuria and nephrocalcinosis in a middle-aged man. AB - We report a 41-year-old man with hypomagnesemia, hypercalciuria, nephrocalcinosis, myopia and horizontal nystagmus. The hypomagnesemia was due to primary renal magnesium loss. He was diagnosed as having the syndrome of renal hypomagnesemia, hypercalciuria and nephrocalcinosis. This is a rare condition generally diagnosed by the first to third decades of life. Renal failure is common and end-stage renal disease can occur in children or young adults. The patient was treated with oral magnesium, chlorthalidone, potassium citrate and allopurinol and was followed up for 3 years. Treatment resulted in an improvement in hypercalciuria but serum magnesium level could not be normalized. The patient's renal function remains stable, with a mild degree of renal insufficiency. PMID- 12745756 TI - The Norwegian welfare state on the threshold of a new century. PMID- 12745757 TI - Burnout and self-perceived health among Finnish psychiatrists and child psychiatrists: a national survey. AB - AIMS: This study investigated burnout and related health matters among a sample of Finnish physicians working within the field of psychiatry. METHODS: A postal questionnaire was sent to one in three licensed physicians randomly selected from the register of the Finnish Medical Association (FMA). The response rate was 74% (n = 3133). RESULTS: Psychiatrists and child psychiatrists reported burnout, threat of severe burnout, depression, and mental disorder more commonly than other physicians. Moreover, psychiatrists and child psychiatrists reported less often "good" or "rather good" self-perceived health. Depression had a moderate positive correlation with overall MBI score. Lack of possibilities to consult a colleague, and supervision of work, experience of threat of violence, and self reported depression were significantly associated with overall burnout level and emotional exhaustion. CONCLUSIONS: Emotional exhaustion as a symptom of burnout was common among psychiatrists, especially among those working in community care, and child psychiatrists. Problems of general health, as well as mental health, among psychiatrists and child psychiatrists are in need of attention. PMID- 12745759 TI - Functional ability in the elderly Finnish population: time period differences and associations, 1985-99. AB - AIMS: The aim of this study was to determine whether functional ability among 65- to 79-year-old Finnish men and women changed at population level from the 1980s to 1990s and how gender, age, previous occupation, and marital status are associated with functional ability and whether these associations have changed over time. METHODS: Biennial surveys on health behaviour among Finnish elderly people were used to study 10,309 men and women from 1985 to 1999. Age, gender, previous occupation, and marital status were the demographic variables. Logistic regression was used to determine differences. RESULTS: Functional ability deteriorated clearly with age, but was slightly better in the 1990s than 1980s. Gender differences of functional ability were small. Retired office employees had the best functional ability. Moreover, time changes of functional ability between occupational groups were a little more positive among men than among women. Functional ability of divorced and widowed elderly emerged as slightly worse than among married persons. CONCLUSIONS: Young age and non-manual occupation prior to retirement were associated with better functional ability. The overall improvement of self-reported functional ability among elderly people suggests that the onset of disabilities could be postponed, especially if health-related circumstances were more evenly distributed at the start of or even before retirement age. PMID- 12745758 TI - Blood pressure among the Inuit (Eskimo) populations in the Arctic. AB - AIMS: Studies of blood pressure among various Inuit (Eskimo) populations in the Arctic have given inconsistent results. Most studies reported lower blood pressure among the Inuit as compared with the predominantly white national populations. This has been attributed to traditional subsistence practices and lifestyle. This study compared the blood pressure among the major Inuit population groups with other populations and examined the associations with factors like age, gender, obesity and smoking. METHODS: The study comprised four Inuit populations from Alaska, Canada, and Greenland with participation rates ranging from 51% to 73%. In a cross-sectional design, 2,509 randomly selected adults from 31 villages were examined. Blood pressure, anthropometric measurements, smoking, and medication were recorded. RESULTS: Mean systolic blood pressures ranged from 116 to 124 mm Hg among men and 110 to 118 among women in the four populations. Mean diastolic blood pressures ranged from 75 to 78 mm Hg among men and from 71 to 73 among women. Systolic blood pressure increased with age. Male gender, obesity, being a non-smoker, and being on anti-hypertensive treatment were associated with high systolic and diastolic blood pressure. Adjusted for age, body mass index, smoking, and anti-hypertensive treatment, blood pressure differed among the populations (p /= 6 months) and severe dyspareunia in a non-patient population of women, and to explore the rate of recovery as well as the inclination to seek medical care. Another aim was to compare the use of oral contraceptives among women who had ever had dyspareunia and those who had not. METHODS: A total of 3,017 women aged 20-60 participating in a screening program for cervical cancer answered a questionnaire about possible painful coitus. RESULTS: The prevalence was 9.3% for the whole group and 13% for women aged 20-29 and 6.5% for the women aged 50-60, with a risk ratio of 2.0 (95% CI 1.4-2.8) for the youngest age group compared with the oldest. The incidence risk ratio was 9.3 (95% CI 2.8-30.9) for the youngest age group compared with the oldest. Using age specific incidence rates, a rising incidence of dyspareunia in young women was demonstrated. Of the women who had ever had prolonged and severe dyspareunia 28% had consulted a physician for their symptoms; 20% recovered after treatment, while 31% recovered spontaneously. No differences were found in the use of oral contraceptives between the women who had had dyspareunia and those who had not. CONCLUSIONS: Prolonged and severe dyspareunia is a great health problem among all women and especially young women, for whom a rising incidence of dyspareunia is suggested and discussed. Surprisingly few women have consulted a physician, raising the question of why this is the case and what can be done about it. PMID- 12745762 TI - Cost of dementia: impact of disease progression estimated in longitudinal data. AB - AIMS: Several studies have shown that health care costs are higher for demented than for non-demented persons and that health care costs are higher for more severe demented persons than less severe demented persons. However, most studies report on cross-sectional study designs, and thus fail to examine the influence of disease progression on changes in health care costs to individual persons. The objective of this study was, using longitudinal data, to examine changes in total health care costs with disease progression in demented persons. METHODS: We assumed that disease progression could be characterised by transitions between different states of dementia which reflected the degree to which the disease progressed over time. Then, changes in health care costs were regressed on a set of explanatory variables including disease progression. A total of 465 demented and non-demented persons were interviewed twice. The time between interviews was about three years. Before each interview, the participant was examined for dementia and classified by type of dementia (Alzheimer's disease, vascular or other types of dementia) and degree of dementia (very mild, mild, moderate, severe). RESULTS: The results of this longitudinal study confirmed that health care costs increased over time for non-demented as well as for demented persons and that health care costs increased with disease progression. In particular, the health care costs increased when the disease had progressed into the severe state of the dementia. Also, decline in functional abilities was an important factor for explaining changes in health care costs. PMID- 12745763 TI - Social position and health in old age: the relevance of different indicators of social position. AB - AIMS: An analysis was undertaken to investigate social inequalities in health among old men and women in relation to five indicators of social position. METHODS: The study is based on a population-based cross-sectional survey among 748 75-year-old men and women, which was performed as clinical examinations and interviews in 1989 in Glostrup, a suburban area west of Copenhagen. Social position was measured by vocational education, occupation, social class, income, and housing tenure. Health was measured by number of chronic diseases, tiredness in relation to mobility, need of help in relation to mobility, oral health (number of teeth), and well-being (the CES-D Scale). The statistical analysis included bivariate contingency tables and logistic regression analyses. RESULTS: Two material wealth variables (income and tenure) were consistently related to nearly all health measures while the relationships between the other social position variables and health showed no consistent patterns. Multiple logistic regression analyses with tenure and income as independent variables and each of the health variables as dependent variables and control for education and occupation showed different patterns for men and women. In men the odds ratios of housing tenure on four health variables were strong and unaffected by education and occupation while in women the odds ratios of income on three health variables were strong and unaffected by education and occupation. CONCLUSION: This study demonstrates strong, consistent associations between variables of material wealth indicators and various measures of health among 75-year-old men and women. PMID- 12745765 TI - Women's experiences of long term sickness absence: implications for rehabilitation practice and theory. AB - BACKGROUND: In most European countries, spells of long-term absence contribute the largest number of days that are reimbursed as a result of sickness absence. This group is growing and it is constituted mainly of women. AIM: The present study seeks further knowledge about what happens then and there, i.e. how women on long-term sickness absence handle and explain, for themselves and others, this interruption in their daily life. METHODS: Semi-structured interviews were performed with 82 middle-aged women with personal experience of long-term sickness absence. RESULTS: The women's accounts of sickness absence contained interpretations of what had happened to them, how things were at present, and what they thought the future would bring. Three different accounts could be distinguished: crisis, breakpoint, and migration. The perception of their own situation and especially what they thought about their future was associated with their feeling of power to take the initiative, and their well-being. CONCLUSION: From this study the authors have found implications for central topics of importance: time elapse, sense of coherence, reorientation/adaptation, vital goals, and gender. PMID- 12745764 TI - Risk factors of perinatal mortality in Lithuania, 1997-1998. AB - AIM: The aim of this paper is to determine the risk factors of perinatal mortality in Lithuania during 1997-1998. METHODS: The focus of the study is the 1997-1998 Medical Birth Register Database (N = 75,178) covering social demographic factors, the obstetric anamnesis of previous pregnancies, as well as mothers health behaviour and pathology of the current pregnancy, delivery and obstetric aid. The multivariate analysis assessing odds ratios of risk factors for perinatal mortality using logistic regression has been carried out and related population attributable risk fractions were calculated. RESULTS: A statistically significant (p < 0.01) model predicting perinatal mortality has been obtained in which 19 out of 76 analysed factors have been distinguished to be statistically significant. The septic state of the mother during the delivery (OR = 13.3; CI = 8.9-19.9) and haemorrhage due to a premature placental separation (OR = 11.6; CI = 7.9-16.9) were associated with substantial increased risks of the foetus. Factors from other groups (perinatal pathology during previous pregnancies and social, behavioural or environmental factors) had somewhat lower, though significant, risk on perinatal mortality. The highest population attributable risk fraction (PARF) was found for the breech birth and foetal growth retardation (PARF = 13 percent and PARF = 9.7 percent, respectively). It should be pointed out that the impact of marital status on perinatal mortality was very high (PARF = 6.4 percent), even higher than that of some clinical factors, though its odds ratio was among the lowest (OR = 1.5; CI = 1.2-1.7). CONCLUSION: During 1997-1998, the highest risk for perinatal mortality in Lithuania was associated with obstetric aid during delivery and pathology of current pregnancy. These groups of risk factors had also the highest population attributable risk fraction. PMID- 12745766 TI - How does individual smoking behaviour among hospital staff influence their knowledge of the health consequences of smoking? AB - AIMS: This study examined associations between individual smoking habits among hospital staff and their knowledge of the health consequences of smoking and passive smoking. The a priori hypothesis was a higher level of knowledge among non-smokers compared with smokers. METHODS: A survey was undertaken, based on self-administered questionnaires at a Danish hospital (Frederikssund Hospital) in the Copenhagen area. Descriptive statistics, chi-square test and multiple logistic regression analyses were used. A backward stepwise elimination of variables at a 5% level of significance was performed and 95% confidence intervals were calculated. Main outcome measures were knowledge of the health consequences of smoking, passive smoking and other lifestyle factors. RESULTS: A total of 445 of 487 employees (91%) from all professional groups returned the questionnaire. Compared with ex- and never smokers, smokers systematically underestimate the health consequences of smoking and passive smoking independent of profession, department, sex, and age. There is no consistent association between knowledge of the health consequences of smoking and profession and department. There are significant inverse associations between smoking and knowledge of the health effects of excess use of alcohol and lack of physical activity. CONCLUSION: Individual smoking habits among hospital staff strongly influence smoking-related knowledge. No other variables are of consistent importance. These findings are supported by the literature. The validity of the study is good, but a similar study in a bigger population would strengthen the evidence. PMID- 12745767 TI - Hard choices in public health: the allocation of scarce resources. PMID- 12745773 TI - Measuring the mental health status of the Norwegian population: a comparison of the instruments SCL-25, SCL-10, SCL-5 and MHI-5 (SF-36). AB - A great number of questionnaires and instruments have been developed in order to measure psychological distress/mental health problems in populations. The Survey of Level of Living in 1998 conducted by Statistics Norway used both Hopkins Symptom Checklist (SCL-25) and the Short Form 36 (SF-36), including the five-item mental health index (MHI-5). Five-item and 10-item versions of the SCL-25 have also been used in Norwegian surveys. The purpose of this study was to investigate the correlation between the various instruments, and to assess and to compare psychometric characteristics. A random sample of 9735 subjects over 15 years of age drawn from the Norwegian population received a questionnaire about their health containing SCL-25 and SF-36. Response rate was 71.9%. Reliability of the SCLs and MHI-5 were assessed by Cronbach alpha. The scores from full and abbreviated instruments were compared regarding possible instrument-specific effects of gender, age and level of education. The correlations between the instruments were calculated. The capacity of the various instruments to identify cases was assessed in terms of sensitivity, specificity, predictive values, receiver operating characteristics (ROC) and area under the curve (AUC). The reliabilities were high (Cronbach alpha>0.8). All instruments showed a significant difference in the mean scores for men and women. The correlation between the various versions of SCL ranged from 0.91 to 0.97. The correlation between the MHI-5 and the SCLs ranged from -0.76 to -0.78. The prevalence rate was 11.1% for SCL-25 scores above 1.75 and 9.7% for scores below 56 in MHI-5. AUC values indicated good screening accordance between the measures (AUC>0.92). The results suggest that the shorter versions of SCL perform almost as well as the full version. The corresponding cut-off points to the conventional 1.75 for SCL 25 are 1.85 for SCL-10 and 2.0 for SCL-5. MHI-5 correlates highly with the SCL and the AUC indicate that the instruments might replace each other in population surveys, at least when considering depression. An operational advantage of the MHI-5 over the SCL instruments is that it has been widely used not only in surveys of mental health, but also in surveys of general health. PMID- 12745774 TI - The Hopkins Symptom Checklist-25 in screening DSM-III-R axis-I disorders. AB - Simple screening questionnaires for major psychiatric disorders are needed for epidemiological research and clinical work. We describe the characteristics of the Hopkins Symptom Checklist-25 (HSCL-25) as a screening instrument in a two phase epidemiological survey using the Structured Clinical Interview for DSM-III R (SCID) as a diagnostic tool. The material consisted of 1609 subjects aged 31 years who were asked to participate in a health survey. The invitation included the HSCL-25 questionnaire. All "screen-positive" (HSCL-25 mean>/=1.55) subjects and every tenth "screen-negative" subject were invited to participate in the SCID interview. The sensitivity of the HSCL-25 for any present DSM-III-R axis-I psychiatric disorder was 48%. The specificity was 87%. The sensitivity of cases with comorbid psychiatric disorders was 100%. The HSCL-25 is a moderate instrument for screening with present axis-I DSM-III-R psychiatric disorders in a young adult population. It can be recommended for screening of psychiatric disorders. PMID- 12745775 TI - The long-term stability of temperament traits measured after a suicide attempt. A 5-year follow-up of ratings of Karolinska Scales of Personality (KSP). AB - The main aim of the present study was to investigate whether or not temperament dimensions are stable over time. Twenty-six patients (21 women and five men) filled in the Karolinska Scales of Personality (KSP) both at admission after a suicide attempt (index) and at follow-up 5 years later. KSP changes were significantly associated with low severity of psychiatric symptoms and no reported reattempts at follow up. There were significant changes of all five groups of KSP, most prominently in anxiety-related scales. At index, KSP scores did not differ between those who later would repeat a suicide attempt (repeaters) and not, but repeaters more often tended to have a personality disorder and their 24-h urinary cortisol tended to be lower. In this limited sample, repeaters seem to have a protracted high anxiety level as mirrored by high and stable KSP scores over time. PMID- 12745776 TI - Suicidal ideation and behavior in children's homes. AB - The aim of this cross-sectional study is to report the self-destructive and suicidal behavior of 98 children and adolescents in child welfare institutions. The children were evaluated using the Child Behavior Checklist (CBCL), the Children's Global Assessment Scale (CGAS) and questionnaires about suicidal and violent behavior, filled in by the child's key worker. Thirty-two per cent of the sample had presented suicidal thoughts, threats or suicide attempts during the previous 6 months. Suicidality was associated with low general functioning level (CGAS<61), self-mutilating behavior and violence. Furthermore, suicidal children had significantly higher CBCL total, externalizing, internalizing, anxious depressive and aggressive scores. Children with suicide attempts (8% of the sample) had a significantly higher number of different types of traumatic experiences before the placement and higher somatization syndrome scores compared to children with suicidal ideation or non-suicidal children. PMID- 12745777 TI - Specific parenting problems when adolescents have emotional and behavioural disorders. AB - Specific parenting dimensions were explored of parents with adolescents (11-17 years old) referred to outpatient clinics for behavioural and emotional problems. Eighty-seven parent-adolescent pairs participated. Parents completed a parenting interview, and the adolescents were administered a diagnostic interview, the Child Assessment Schedule. Parents of adolescents with behaviour disorders showed impairment in 8/10 parenting dimensions compared to parents of adolescents with emotional disorders or parents of adolescents with no disorder. Parents with behaviour-disordered adolescents had more problems with Appreciation, Involvement, Setting Limits, Consistency, Monitoring Contact, Parenting Priority and Negative Attributions than did other parents. Impaired parenting by the same gender parent was more related to the adolescent's problems than the opposite gender parent. Fathers' lack of contact with their sons strongly correlated with their sons' expression of anger. PMID- 12745778 TI - Parental rearing and individual vulnerability to drug addiction: a controlled study in a Swedish sample. AB - In a convenience sample of 81 healthy subjects vs. a group of 81 heroin addicts from the Methadone program in Stockholm, Sweden, the hypothesis of a possible link between experiences of dysfunctional parental rearing and the subsequent development of dysfunctional assumptions concerning self and others was tested. The subjects (n=162) completed the EMBU to report perceptions of parental rearing behaviour, two measures of dysfunctional assumptions and dysfunctional working models, the Dysfunctional Attitude Scale (DAS) and the DWM-S, and the Screening Drug Career Questionnaire (SDCQ) for assessing different aspects of the drug career in the sample of addicts. Four predictions were made: 1) parental emotional warmth should be negatively correlated with DAS and DWM-S scores; 2) experiences of dysfunctional parental rearing activities should be correlated with high scores on the DAS and the DWM-S; 3) there should be significant differences between the group of addicts and the control group on the EMBU first order factors Rejection, Emotional warmth, Overprotection, and on the EMBU midparent subscale and finally 4) the results should support the hypothesis of an overprotecting mother and a rejecting father. The four predictions were supported by the results. The inter-group differences in quality of rearing shown in this study support the assumption of the impact of parental rearing on the development of dysfunctional working models of self and others. Accordingly, the presented data confirm that a parental rearing behaviour perceived both as Rejecting and Overprotective represents a link between dysfunctional parenting and the development of maladaptive psychosocial behaviour like drug addiction. PMID- 12745785 TI - Child and adolescent psychiatric disorders predicting adult personality disorder: a follow-up study. AB - The objective of this study was to examine associations between childhood and adolescent psychiatric disorders and adult personality disorders in a group of former child psychiatric inpatients. One hundred and fifty-eight former inpatients with a mean age of 30.5 +/- 7.1 years at investigation had their childhood and adolescent Axis I disorders, obtained from their medical records, coded into DSM-IV diagnoses. Personality disorders in adulthood were assessed by means of the DSM-IV and ICD-10 Personality Questionnaire (DIP-Q). The predictive effects of child and adolescent Axis I disorders on adult personality disorders were examined with logistic regression analyses. The odds of adult schizoid, avoidant, dependent,borderline and schizotypal personality disorders increased by almost 10, five, four, three and three times, respectively, given a prior major depressive disorder. Those effects were independent of age, sex and other Axis I disorders. In addition, the odds of adult narcissistic and antisocial personality disorders increased by more than six and five times, respectively, given a prior disruptive disorder, and the odds of adult borderline, schizotypal, avoidant and paranoid personality disorders increased between two and three times given a prior sub-stance-related disorder. The results illustrate an association between mental disorders in childhood and adolescence and adult personality disorders. Identification and successful treatment of childhood psychiatric disorders may help to reduce the risk for subsequent development of an adult personality disorder. PMID- 12745786 TI - The relationship of needs and quality of life in persons with schizophrenia living in the community. A Nordic multi-center study. AB - The relationship between needs for care and support and subjective quality of life was investigated in a cross-sectional multi-center study including 418 individuals with schizophrenia from 10 centers in Nordic countries. Needs in 22 domains were investigated by interviews with key workers and their patients using the Camberwell Assessment of Need scale, and quality of life by the Lancashire Quality of Life Profile. The results showed that key workers rated slightly more needs than patients. To have more unmet needs, as rated by both key workers and patients, were correlated to a worse overall subjective quality of life, while met needs showed no such association. A regression analysis, controlling for clinical and social characteristics of the patients, showed more unmet needs to be associated with a worse quality of life, accounting for 6% out of a total of 41% explained variance in subjective quality of life. Regression analyses of the relationship of unmet needs in specific life domains and overall quality of life showed that unmet needs in five domains as perceived by patients accounted for 17% of the explained variance in overall quality of life. More than half of this variance was related to an unmet need in the domain of social relationships. It is concluded that unmet needs are of specific importance in needs assessment and that attention must be paid to separate met needs for care and services from unmet needs, since the latter seem more important to consider in order to improve outcome of interventions with regard to quality of life. Specific attention should in this context also be paid to unmet needs concerning social relationships and problems with accommodation. PMID- 12745787 TI - Prevalence of depression among general hospital surgical inpatients. AB - Previous studies report a wide range of prevalence rates of depressive illness among general hospital inpatients, all higher than in a non-patient population. Several factors may have influenced on these results. Mixed study population, depression-prone subgroups and continuous shift in what is a surgical inpatient population due to day surgery treatment are examples. In the present study, 108 patients were assessed with the Structured Clinical Interview for DSM-III-R Non Patient (SCID-NP) version for current major depression (CMD) and for dysthymia. Furthermore, a patient self-rating scale for depressive symptoms and anxiety, the Hospital Anxiety and Depression Scale (HADS), was applied. CMD was diagnosed among 14/108 patients (13%). Depressive symptoms (HADS-D> or =8) were seen in 14 patients. Symptoms of anxiety (HADS-A> or =8) were seen in 12/14 CMD patients (86%). Ten of the 14 patients diagnosed as CMD (71%) did not receive any pharmacological antidepressant treatment. After excluding known depression-prone subgroups of patients representing a bias, this study showed that in a community hospital group of adult surgical patients between 18 and 65 years of age, the prevalence of depression is still somewhat higher than in the general population, but not as high as in the previous studies on general hospital patients to which we referred. Thus, this high prevalence of depression in part could be due to increased depression rates in certain population subgroups. However, this hypothesis alone is not sufficient to explain the present results fully. PMID- 12745788 TI - The Finnish version of the Dissociative Experiences Scale-II (DES-II) and psychiatric distress. AB - The confirmatory factor structure of the Finnish version of the Dissociative Experiences Scale II (DES-II) was exactly the same in the non-clinical population (n=924) as was found in the most recent study in the USA (with the original English version) and as suggested by the authors of the original version. The Finnish version of the DES-II has a Cronbach's alpha of 0.92, and there is a clear positive correlation (from 0.31 to 0.66) between all 28 items and the total score (P<0.001 for all items), indicating good internal consistency and reliability. We also investigated dissociative experiences and psychiatric distress with the DES-II and the General Health Questionnaire (GHQ-12). The increased proportion of "pathological dissociation" (DES-T) to "non-pathological dissociation" (DES-NP) was directly linked with increased distress. This finding is meaningful for the debate on the utility of the different DES-II scoring systems. PMID- 12745789 TI - Personality dimensions measured by the Temperament and Character Inventory (TCI) in subjects with social phobia. AB - The present study investigated personality dimensions by means of the Temperament and Character Inventory (TCI) in subjects with social phobia with or without a co existing avoidant personality disorder. Thirty-one individuals with social phobia were recruited through advertisement and diagnosed with the Structured Clinical Interview for DSM-IV I and II psychiatric disorders. Comorbid Axis I psychiatric disorders were diagnosed in 29% of the cases. Co-occurring personality disorders were present in 55.8% of the cases, and avoidant personality disorder in 48.4%. The social phobia subjects, as compared to healthy controls of the same age, scored significantly higher in the TCI dimension measuring Harm avoidance but significantly lower in Persistence, Self-directedness, Cooperativeness and Self transcendence. Presence of avoidant personality disorders in the social phobia subjects was associated with significantly higher Harm avoidance, particularly on the subscale Shyness with strangers. In conclusion, individuals with social phobia were characterized by high comorbidity of avoidant personality disorder and deviations in TCI personality dimensions. Enhanced Harm avoidance was the most prominent personality trait. The observed deviations in TCI dimensions were primarily related to the social phobia itself and not to the presence of concurrent personality disorders. PMID- 12745790 TI - A new priority in psychiatry: focused services for adolescents. AB - Finland was the first European country to give a high priority to the development of specialized psychiatric services for adolescents. The aim of this paper is to 1) describe the development of these services, 2) study the status and the functioning of psychiatric services for adolescents in 2000 and 3) present some future challenges. The data have been collected from the social welfare register, from a questionnaire sent to Health Districts and via telephone interviews. In addition, the functioning of the treatment system has been studied in detail in one Health District by means of interviews and questionnaires. The general targets set in 1987 giving national priority to focused psychiatric services for adolescents were mostly reached in 2000, and resources for adolescents have increased markedly. In spite of this, minors still have to be treated in adult psychiatric wards. Many problems were detected in the functioning of the chains of treatment for adolescents. The differentiation between primary and secondary care was unclear. Primary care had a limited capacity in the early detection of mental disorders in adolescents. The services for adolescents were broken down into small units. Furthermore, there were no comprehensive future plans. The need for specialized psychiatric services for adolescents is evident. In addition to developing adequate resources, the quality of care should also be guaranteed by training and by competent long-term planning of psychiatric service systems. PMID- 12745791 TI - Psychiatric staff members' emotional reactions toward patients. A psychometric evaluation of an extended version of the Feeling Word Checklist (FWC-58). AB - The study evaluated the psychometric properties of an extended version of the Feeling Word Checklist (FWC-58), which measure the therapists' emotional reactions toward patients. Doctors, psychologists, nurses and aides from 23 wards in seven widely different psychiatric departments completed a total of 3012 forms. The original Feeling Word Checklist (Whyte CR, Constantopoulos C, Bevans HG. Br J Med Psychol 1982;55:187-201) was expanded with 28 feeling words covering more feelings of being invaded, idealized, devalued and of being secure. The rating scale was changed from a two-point (yes/no) scale to a five-point scale ranging from not at all (=0) to very much (=4). The factor analysis revealed ten factors with an eigenvalue equal or greater than 1.0. The two first factors were clearly the strongest but a two-factor solution did not satisfactorily reflect the data. The scree test indicated four to seven factors. We chose a seven-factor solution, as this seemed clinically most meaningful. Based on the factors we developed seven indices which were named: Important, Rejected, Confident, Bored, On guard, Overwhelmed and Inadequate. The subscales had satisfactory internal consistency and described meaningful emotional profiles of the different psychiatric wards and the individual patients. The seven subscales seem well worth to use in further research and in a clinical context. PMID- 12745792 TI - The role of the sympathetic nervous system in anxiety: Is it possible to relieve anxiety with endoscopic sympathetic block? AB - The function of the autonomic nervous system is divided so that the parasympathetic system spares central nervous system energy and the sympathetic system makes extra energy available and consumes it. The sympathetic nervous system then prepares our body for emergency and it always functions when our conscious or even unconscious mind notices a need for defence or to provide energy. A surgical procedure, where the upper thoracic sympathetic ganglions are ablated, either with cauterization or clamping with metallic clips, has been used to treat sweating of the hands and facial blushing for decades. Instead of ablating large areas of sympathetic trunk, which can cause severe side-effects such as reflex sweating of the body, the surgical procedure is nowadays carried out in a more precise symptom-mediating level of uppermost thoracic sympathetic ganglia. Blushing, hyperhidrosis of palms and head, and trembling are common in social phobia, and they seem to be provoked by the activation of the sympathetic nervous system. Preliminary studies show that some social phobia patients may benefit from the endoscopic sympathetic block (ESB). If the patient with generalized social phobia has not received help with adequate medication or psychotherapy, the ESB may be a new possible treatment of choice. PMID- 12745793 TI - Chronic pain disorder associated with psychogenic versus somatic factors: a comparative study. AB - Fifty-one consecutive non-depressed patients with chronic pain referred to a multidisciplinary pain clinic were assessed. In 32 patients, pain was judged to be associated with psychogenic factors only, while pain in 19 patients could be attributed solely to a general medical condition. The methods of investigation comprised visual analogue scales (VAS) and pain drawings, the Dysfunctional Attitude Scale (DAS), the Karolinska Scales of Personality (KSP), locus of control (LOC) and EMBU (for assessing perceived parental rearing practices). Mean age, gender distribution, analgesic consumption, pain duration, percentage of body area pain as well as body localization of pain were comparable in both groups. Patients with psychogenic pain reported higher levels of general bodily discomfort but less concentration difficulties and memory disturbances compared with the somatic pain patients. There were no significant intergroup differences on any of the LOC, DAS, KSP or EMBU items. LOC turned out to be extremely external, whereas DAS, KSP and EMBU scores were comparable to normal controls in earlier work. The paucity of differences between the two patient groups and the unremarkable personality structure of patients led to somewhat conflicting conclusions, and the results of the study pose one more piece of evidence for the futility of the dichotomous organic vs. psychogenic distinction of chronic pain disorders. PMID- 12745796 TI - Women--a neglected risk group for atherosclerosis and vascular disease. AB - Women are a neglected group for cardiovascular disease. Whereas young women tend to have lower incidences of coronary artery disease, stroke and myocardial infarctions than men, the situation changes drastically at menopause, after which women are at greater risk than men. Despite this, women at all ages receive less treatment, less attention and not enough information about health risks. Most risk factors, e.g. hypertension, elevated blood lipid levels, diabetes and changes in oestrogen levels, differ between women and men. As a consequence of this, secondary prevention from coronary artery disease is likely to have different effects in women to those in men. Different kinds of antihypertensive therapy, hormone-replacement therapy and lipid-lowering treatment may be more or less suitable in women than in men. The recent development of angiotensin receptor blockers may have beneficial effects which make them particularly effective in women. PMID- 12745797 TI - Microdialysis for neurochemical monitoring of the human brain. PMID- 12745798 TI - The clinical impact of vascular growth factors and endothelial progenitor cells in the acute coronary syndrome. PMID- 12745799 TI - Serum myoglobin/carbonic anhydrase III ratio in the diagnosis of perioperative myocardial infarction during coronary bypass surgery. AB - OBJECTIVE: The purpose of the present study was to evaluate the usefulness of the myoglobin/carboanhydrase III (Myo/CAIII) ratio in the diagnosis of perioperative myocardial infarction during coronary artery bypass surgery. DESIGN: Thirty patients undergoing elective coronary artery bypass grafting (CABG) were included in the series. The patients were randomized in two groups: one received conventional normothermic retrograde blood cardioplegia, while the other was subjected to a 5-min period of ischemic preconditioning before cardioplegia. Biochemical markers for myocardial and skeletal muscle injury were measured in serial blood samples taken postoperatively from 4 h after aortic declamp. RESULTS: Three patients were diagnosed to have suffered from perioperative myocardial infarction on the basis of significant elevations of troponin T and creatine kinase MB-isoenzyme (CK-MB) concentrations. In these particular patients the Myo/CAIII ratio increased rapidly after aortic declamping. In uncomplicated patients, the median value of the Myo/CAIII ratio remained within normal limits. There was a positive correlation between the net output of lactate during the aortic cross-clamping period and postoperative Myo/CAIII ratio. The Myo/CAIII ratio proved to be a more specific indicator for myocardial damage than myoglobin alone. The Myo/CAIII ratio was higher in the preconditioning group than in the control group. CONCLUSION: Myo/CAIII ratio is a sensitive and specific marker for perioperative myocardial infarction increasing rapidly after aortic declamping. This ratio could also be used when assessing the extent of ischemic myocardial injury and comparing different surgical and cardioprotective techniques. PMID- 12745800 TI - Endothelial damage after treatment with low-molecular weight heparins--a morphological study. AB - OBJECTIVE: Recent studies failed to show long-term benefit with low-molecular weight heparins (LMWH) in unstable coronary heart disease. A previous study of vascular effects of the cytostatic agent 5-fluorouracil (5-FU) showed that dalteparin prevented thrombosis induced by 5-FU but endothelial damage was not ameliorated and was present also in animals treated with dalteparin only. This study investigates the influence of LMWH currently in clinical use on arterial endothelium in vivo. DESIGN: Eighty rabbits in four groups were treated with dalteparin, enoxaparin, tinzaparin and saline, respectively. Arterial endothelium was examined after 3, 14, 30 and 60 days with scanning electron microscopy. RESULTS: All three groups treated with LMWH showed moderate damage to the endothelium, with contracted vessel wall and endothelial cells, cell membrane damage, denudation of subendothelium and adhering platelets. Contrarily, the control group exhibited a normal endothelium. CONCLUSION: Morphologic examination of arterial endothelium shows that all investigated LMWH exert a moderate toxic effect on endothelial cells. The clinical impact of these observations, e.g. concerning effect of long-term LMWH treatment, needs to be further elucidated. PMID- 12745801 TI - Adventitial myofibroblasts play no major role in neointima formation after angioplasty. AB - OBJECTIVE: Myofibroblasts migrating from adventitia have been suggested to constitute a majority of neointimal cells after angioplasty. We sought to examine this hypothesis by use of smoothelin, which is a marker for the quiescent smooth muscle cell (SMC) phenotype while not expressed by myofibroblasts. DESIGN: Balloon angioplasty was performed in left iliac arteries of 25 rabbits that were killed after 3-56 days. Arterial cross-sections were immunostained for alpha actin (general marker), smoothelin (quiescent SMC phenotype), and Ki-67 (proliferative phenotype). RESULTS: Adventitial cells became transiently actin positive (myofibroblasts) but did not express smoothelin at any time point. In media, angioplasty induced transient proliferation and coinciding transient decrease in smoothelin expression. Neointimal cells, present 7 days after angioplasty, were initially proliferating and smoothelin-negative but changed to non-proliferating, smoothelin-positive cells after 56 days where 82 +/- 10% of cells stained positive for smoothelin. This phenotypic modulation of medial and intimal cells began in media and moved gradually towards the lumen. CONCLUSION: At late follow-up, the majority of intimal cells are smoothelin-positive indicating that adventitial myofibroblasts play no major role for neointima formation. PMID- 12745802 TI - Conduction properties of accessory atrioventricular pathways: importance of the accessory pathway location and normal atrioventricular conduction. AB - OBJECTIVE: The objective of this study was to determine the conduction properties of the normal atrioventricular (AV) conduction system in relation to accessory pathway (AP) location in patients with symptomatic Wolff-Parkinson-White syndrome. DESIGN: The conduction properties of the AP and the AV node were studied in 356 patients with single manifest AP who underwent successful ablation. RESULTS: Sixty-three percent of the APs were located on the left free wall (226) and the remaining 37% were posteroseptal, anteroseptal and right free wall. AV block (PR > or =220 ms) was observed in 15 patients, 8 cases being associated with the right free wall (15%) compared with 7 on the left free wall (3%, p = 0.002) and none on the anteroseptal (p = 0.02) or posteroseptal (p = 0.007). The PR (182 +/- 30 ms) and AH (102 +/- 25 ms) intervals associated with right free wall AP were longer than left free wall (166 +/- 23, 88 +/- 21 ms, p < 0.0001, respectively), anteroseptal (155 +/- 11, 79 +/- 12 ms, p < 0.0001) and posteroseptal AP (155 +/- 16, 79 +/- 13 ms, p < 0.0001), whereas the PR and AH intervals associated with posteroseptal and anteroseptal AP were shorter than left free wall AP (p < 0.05). When patients with AV block were excluded from the analysis, the PR intervals in patients with right free wall AP were still longer than in those with left free wall (p < 0.005), anteroseptal (p < 0.001) and posteroseptal AP (p < 0.0001), as were the PR intervals associated with left free wall AP compared with data of posteroseptal (p < 0.01) and anteroseptal AP (p < 0.05). Significant differences in AV nodal effective refractory period, anterograde and retrograde AV block cycle length were also observed in relation to AP location. CONCLUSION: The conduction properties of the normal AV conduction system are associated with specific AP location in patients with Wolff-Parkinson White syndrome. PMID- 12745803 TI - Low prevalence of ischemic electrocardiographic findings in a Danish population. AB - OBJECTIVE: Mortality and incidence of cardiovascular disease have declined during the past 35-40 years. The dual aim of this study is to investigate whether the prevalence of electrocardiographic findings is low compared with older studies and to describe the prevalence of electrocardiographic findings in the Danish population, which has not been reported since 1981. DESIGN: Cross-sectional study based on electrocardiograms obtained from a random sample of the population in the district of Ebeltoft, Denmark, December 1991-June 1992. RESULTS: The age and sex stratified prevalence of abnormal electrocardiograms ranged from 6.8% (95% CI: 4.01-10.7%) in women to 15.0% (95% CI: 10.6-20.4%) in men aged 41-51 years. Men had significantly more electrocardiographic changes than women (p = 0.004). Frequent findings were signs of earlier myocardial infarction (3.1%; 95% CI: 2.1 4.5%), axis deviation (3.1%; 95% CI: 2.1-4.5%) and incomplete right bundle branch block (1.2%; 95% CI: 0.6-2.2%). CONCLUSION: Prevalence of ischemic electrocardiographic findings is low when compared with studies from the past 50 years. PMID- 12745804 TI - Decreased lung function and exercise capacity in Fontan patients. A long-term follow-up. AB - OBJECTIVE: The haemodynamic situation for patients with Fontan circulation is characterized by a reduced, non-pulsatile pulmonary blood flow. To evaluate if this has any impact on lung function and exercise capacity, we studied the surviving 20 patients operated upon at our institution between 1980 and 1991. The median age was 17.5 years, the median follow-up time was 11.5 years. DESIGN: Lung volumes, flow-volume curves, the ventilatory distribution and the diffusion capacity for carbon monoxide were obtained using routine methods. The exercise tests were performed on a bicycle ergometer with determination of ventilation, oxygen uptake, carbon dioxide production, respiratory rate and heart rate. RESULTS: The lung volumes, maximal expiratory flows and diffusion capacity were significantly lower than expected. The median maximal oxygen uptake was 1.39 for the females and 1.63 l/min for the males, corresponding to 25.1 and 25.2 ml/kg/min, respectively. The maximal heart rates varied from 104 to 177 beats/min. All patients reached a respiratory exchange ratio above 1.0. CONCLUSION: Fontan patients have small lungs. They have a markedly reduced exercise capacity and a low maximal heart rate. PMID- 12745806 TI - Did mothers begin with an advantage? A study of childbirth and maternal health in England and Wales, 1778-1929. AB - This paper contributes to two ongoing debates among demographers. One deals with the immediate and deferred health effects of childbearing in the past, and the other with competing explanations--the frailty and insult accumulation hypotheses -for differences in individual health later in life. The study population consists of working women who lived at four locales in England and Wales in parts of the period 1778-1929 and who were under observation for incapacitating sickness during and after their childbearing years. Mothers within the study population are contrasted with a comparison group made up principally of non mothers. The mothers began their reproductive careers with an advantage in health that was especially evident in the duration of sickness episodes. Even though individual births were less hazardous than individual sicknesses at the same ages, the cumulative effect of childbearing appears to have eroded the mothers' advantage. By ages 50-74 the mothers resembled the comparison group in health. PMID- 12745807 TI - Why are uneducated women in India using contraception? A multilevel analysis. AB - While women's education continues to be strongly associated with lower fertility in India, an important feature of India's current fertility transition is the spread of contraceptive use among uneducated women. Indeed, changes in their fertility are now making the major contribution to the country's overall fertility decline. We use multilevel statistical procedures to investigate the variation in contraceptive use among uneducated women across India. The analysis suggests that, while many of the expected socio-economic variables play their part, there are also considerable diffusion effects in progress, many of which operate at levels beyond the uneducated women's own individual circumstances. For example, we find significant relationships with others' use of contraception and others' education. Mass media exposure also emerges as an important diffusion channel. The multilevel analysis also reveals significant clustering of contraceptive use at different levels, much of which is accounted for by the variables included in the models. PMID- 12745809 TI - Does an effect of marriage duration on pre-transition fertility signal parity dependent control? An empirical test in nineteenth-century Leuven, Belgium. AB - It has been demonstrated for many pre-industrial populations that the age at marriage, or marriage duration, influences age-specific marital fertility but the reason for this remains unclear. Among the several mechanisms that may be responsible, the following are often cited: secondary sterility or increased subfecundity associated with parity; declining coital frequency; the age difference between the spouses; and, importantly, parity-dependent fertility control. If the latter mechanism were partly responsible for the marriage duration effect in pre-transition populations, it would contradict the concept of the modern fertility transition as the evolution (or revolution) from parity independent to parity-dependent fertility. The study presented in this paper investigates the relative importance of these alternative explanations. The application of multivariate Poisson regression to the fertility data from two birth cohorts in the Belgian city of Leuven shows that a linearly declining or even concave age-specific fertility pattern, disaggregated by age at marriage, does not imply parity-dependent fertility limitation. PMID- 12745810 TI - Do parents really matter? Child health and development in Spain during the demographic transition. AB - Linked life histories of children and of their parents living in Aranjuez (Spain) between 1870 and 1950 are used to assess the health and well-being of children in terms of the survival status of their parents. The loss of a mother leads to dramatic increases in the mortality of young children, especially during the first 2 years of life, while the loss of a father has a rather limited negative impact. Over time the relative importance of the loss of a mother increases sharply, thus affording strong, albeit indirect, evidence of their role for mortality reduction during the demographic transition. Heights of military conscripts are used to assess other elements of health unrelated to survival. Results suggest that orphans were noticeably shorter than non-orphans. Over time this effect diminishes thanks to increasingly effective public assistance for orphans. PMID- 12745811 TI - Do unintended pregnancies carried to term lead to adverse outcomes for mother and child? An assessment in five developing countries. AB - This paper investigates whether children later reported as having been unwanted or mistimed at conception will, when compared with children reported as wanted, show adverse effects when the following criteria are applied: receipt of antenatal care before the sixth month of gestation, supervised delivery, full vaccination of the child, and child growth (stunting). The study uses data from five recent Demographic and Health Survey enquiries in Bolivia, Egypt, Kenya, Peru, and the Philippines. In Peru, children unwanted at conception were found to have significantly worse outcomes than other children, but in the other countries, a systematic effect was found only for receipt of antenatal care. Weak measurement of the complex concept of wantedness may have contributed to these results. Birth order of the child, with which wantedness is inextricably linked, has more powerful and pervasive effects, with first-born and second-born children being much less likely to show adverse effects. PMID- 12745817 TI - Neutralization theory and the denial of risk: some evidence from cannabis use among French adolescents. AB - In contemporary societies, risk culture and risk profiling lead to the stigmatization of unhealthy behaviours as 'risky'. Risk denial theory refers to a cognitive way to deal with risky behaviours and can be considered as an updated variant of Sykes and Matza's neutralization theory. People neutralize the 'risky' label using specific techniques that must be added to those previously enlisted by Sykes and Matza. This paper introduces and discusses three techniques of risk denial: scapegoating, self-confidence and comparison between risks. As it is usually defined and studied as a 'risky behaviour', cannabis use provides a relevant example to illustrate these types of risk denial, thanks to various ethnographic studies (including Becker's seminal work on marijuana smokers) and quantitative French data from the 1999 European School Survey on Alcohol and Other Drugs (ESPAD). In order to deny the 'risky' label, cannabis users scapegoat 'hard drugs' users, they emphasize their own ability to control their consumption personally, or they compare cannabis and alcohol risks. The paper concludes with suggestions for further analyses of risk denial. PMID- 12745816 TI - Risk and panic in late modernity: implications of the converging sites of social anxiety. AB - Comparing moral panic with the potential catastrophes of the risk society, Sheldon Ungar contends that new sites of social anxiety emerging around nuclear, medical, environmental and chemical threats have thrown into relief many of the questions motivating moral panic research agendas. He argues that shifting sites of social anxiety necessitate a rethinking of theoretical, methodological and conceptual issues related to processes of social control, claims making and general perceptions of public safety. This paper charts an alternative trajectory, asserting that analytic priority rests not with an understanding of the implications of changing but converging sites of social anxiety. Concentrating on the converging sites of social anxiety in late modernity, the analysis forecasts a proliferation of moral panics as an exaggerated symptom of the heightened sense of uncertainty purported to accompany the ascendency of the risk society. PMID- 12745818 TI - The revival of death: expression, expertise and governmentality. AB - This paper discusses Walter's (1994) assertion that death in the West has recently undergone a revival. In particular it focuses on his claim that this revival is composed of two different strands: a late modern strand and a postmodern strand. The former, according to Walter, is driven by experts who seek to control death, the latter by ordinary people who seek to express their emotions freely. Describing the history and work of Cruse Bereavement Care, the largest bereavement counselling organization in the UK, we question Walter's distinction. We then problematize Walter's suggestion that the revival of death is caused by general social transformations. In contrast we evoke Rose's (1996) work on 'subjectification' and seek to link recent changes in the management of death and grief to permutations in governmental rationality. PMID- 12745819 TI - Mythscapes: memory, mythology, and national identity. AB - In this paper I seek to challenge the dominant modes of conceiving the relationship between memory and national identity, and in so doing offer analysts of nationalism an improved understanding of the dynamics of national identity formation. The concept of collective memory is invoked regularly in attempts to explain the pervasiveness and power of nationalism. I argue that the concept is misused routinely in this context, and instead I employ a 'social agency' approach to theorizing, whereby memory is conceived in a more limited and cogent manner. I argue that it is important to distinguish clearly between memory and mythology, both of which are essential to understanding national identity, for not only are the two concepts distinct, they can also act in opposition to each other. Following from this I introduce the notion of a 'mythscape', the temporally and spatially extended discursive realm in which the myths of the nation are forged, transmitted, negotiated, and reconstructed constantly. Through employing the idea of a mythscape we can relate memory and mythology to each other in a theoretically profitable way. PMID- 12745820 TI - Intermarriage and the demography of secularization. AB - One way of measuring religious affiliation is to look at rites of initiation such as baptism. English statistics show that for the first time since the Church of England was founded, less than half the nation is Anglican on this criterion. The pattern of formal religious transmission changed during the Second World War. Previously christening was quasi-universal, and the Church of England was the preferred provider. By the end of the war baptism was evidently optional, and chosen principally by parents whose religious identities matched. Further analysis suggests that affiliation now tends to be lost following marriage to someone from a different religious background, though the USA differs from Europe in this respect. A demographic theory of advanced secularization is outlined that specifies a proximal cause for declining religious affiliation, and provides tools for predicting the changes to be expected over future decades. The theory also helps to explain why affiliation may fall most quickly where there is most religious diversity. PMID- 12745821 TI - A life-course perspective on social exclusion and poverty. AB - This article assesses whether it is possible to reconceptualize the traditional research approaches to the relationship between poverty and the life cycle on the basis of different sociological perspectives on the life course found in the literature. While the family-cycle approach, which was originally formulated by Seebohm Rowntree (1902), is criticized for being static, descriptive, normative and inflexible, dynamic poverty research is mostly confined to the quantitative analysis of income trajectories, and thus offers only a partial solution to our problem. However, the life-course perspective allows us to combine the best elements of these traditional approaches and to reconceptualize them into a general framework for the study of social exclusion and poverty. To this end, three sociological perspectives on the life course are considered: the traditional North-American life-course perspective formulated by Elder (1974), the Continental institutional approach, and a combined approach which we label the 'political economy of the life course'. Drawing from these three perspectives, we propose a general framework of analysis and formulate hypotheses regarding the phenomena of social exclusion and poverty over the life course which can subsequently be empirically validated. PMID- 12745822 TI - The so-so construction of sociology. PMID- 12745823 TI - Class, generation and Islamism: towards a global sociology of political Islam. PMID- 12745827 TI - A picture is worth a thousand words: children's representations of family as indicators of early attachment. AB - To ascertain whether attachment representations at age 7 are related to early attachment behaviour, family drawings of 123 7-year-olds of known infant attachment status (25 avoidant, 80 secure, 18 resistant) were scored in four ways. Three of these were based in previous attachment research and one was based on a clinical method. The attachment-based coding schemes included specific markers for each attachment pattern (Kaplan & Main, 1985), global ratings (Fury, Carlson, & Sroufe, 1997) and efforts to classify each drawing as belonging to one of the three primary infant attachment groups (secure, avoidant, resistant). In the clinical scheme, children who had been resistant infants were distinguished from the others by use of overlapping and encapsulated figures. For the attachment based schemes, although individual markers were not successful in discriminating attachment groups, the more global approaches (aggregation of markers, global rating scales and judgments of attachment classification) succeeded in this task. In regression analyses controlling for concurrent child and parent measures, infant attachment did not make a significant contribution to predicting insecurity markers in drawings, although child current emotional functioning did. These findings linking attachment relationships with later representations of family relationships were in accord with the conception that avoidant attachment strategies de-emphasize intimate relationships, while resistant attachment strategies are preoccupied with close relationships. These links are most evident in global interpretive strategies rather than those that rely on specific markers. PMID- 12745826 TI - Early temperament and attachment as predictors of the Five Factor Model of personality. AB - To increase our understanding of developmental aspects of the Five Factor Model (FFM) of personality, prospective relations from infant temperament and attachment security to the 'Big Five' dimensions of personality in middle childhood were studied in a sample of 85 Swedish middle class children. Combined maternal and paternal temperament ratings at infant age 20 months and Strange Situation attachment to mother at 15 months were used as predictors of mother and teacher ratings of personality at child age 8 - 9 years. Also the A1 - B2 versus B3 - C2 classification grouping was used as a measure of infant negative emotionality. The results showed extraversion/surgency to be the dimension most clearly related to infancy data; it was predicted by both temperament and attachment security. Attachment security also predicted neuroticism and openness. The emotionality shown in the Strange Situation was not related to the 'Big Five'. The results were discussed in terms of approach and anxiety systems and internal working models as foundations for the FFM personality traits. PMID- 12745828 TI - Measuring sensitivity moment-by-moment: microanalytic look at the transmission of attachment. AB - Meta-analyses have indicated that the mediating power of sensitivity in the transmission of attachment across generations is not nearly as great as would be predicted by attachment theory (van IJzendoorn, 1995; De Wolff & van IJzendoorn, 1997). To make sense of these findings, the author suggests that current measures of sensitivity should be expanded to include the assessment of mother-infant behavior on a micro-behavioral level. This paper reviews evidence for two central propositions: (1) The internal working model of mothers is expressed in subtle, fine-grained interactive behaviors with their infants, and (2) infants perceive and remember these behaviors in the form of interactional expectancies. Following these propositions, hypothesized representational micromodels for each major attachment organization are then offered, based on the Parallel Distributed Processing (PDP) approach to information processing. In closing, this paper presents some of the most pressing unresolved issues for attachment researchers as they seek to empirically identify the mechanisms by which attachment is transmitted across generations. PMID- 12745830 TI - Using attachment theory to understand patients' responses to a therapist's medical break. AB - The impact of a therapist's planned medical break is described and discussed, using attachment theory. The break is compared to Ainsworth's strange situation test and the responses of patients with avoidant, ambivalent and disorganised patterns of attachment are explored. The paper examines patients' responses at the point where news of the break is given and on reunion and looks at communication through breaks in the frame. An overview is given of the understanding of separation and loss in broader analytic theory and the ways in which these complement Bowlby's theory. The clinical cases are reviewed in the light of several authors. PMID- 12745829 TI - Developmental protective and risk factors in borderline personality disorder: a study using the Adult Attachment Interview. AB - Mental representations and attachment in a sample of adults with Borderline Personality Disorder were assessed using the George, Kaplan and Main (1985) Adult Attachment Interview (AAI). Eighty subjects participated in the study: 40 nonclinical and 40 with Borderline Personality Disorder (BPD). The results obtained showed a specific distribution of attachment patterns in the clinical sample: free/autonomous subjects (F) represented only 7%, dismissing classifications (Ds) reached about 20%, entangled/preoccupied (E) 23% and unresolved with traumatic experiences (U) 50%. The two samples differed in their attachment patterns distribution by two (secure vs. insecure status), three (F, Ds and E) and four-way (F, Ds, E and U) categories comparisons. In order to identify more specific protective or risk factors of BPD, 25 one-way ANOVAs with clinical status as variable (clinical vs. nonclinical) were conducted on each scale of the coding system of the interview. Results support the hypothesis that some developmental relational experiences seem to constitute pivotal risk factors underlying this disorder. Results demonstrated potential benefits in using AAI scales in addition to the traditional categories. Implications for research and treatment are discussed. PMID- 12745831 TI - Remembering the forgotten? Reminiscence, hypermnesia and memory for order. AB - Three experiments established that repeated testing affects item and order retention differently: Hypermnesia was found with repeated free recall tests, whereas net performance declined significantly across successive free reconstruction of order tests. Overall order performance declined over tests under a variety of encoding conditions (pictures, words, and relational and item specific processing) and retrieval conditions (intentional and incidental learning). Although net performance dropped across tests, participants did show reliable order recovery (reminiscence) between tests. The implications of these data for general theories of hypermnesia and order are discussed. PMID- 12745832 TI - Stopping and restarting an unfolding action at various times. AB - The ability to inhibit an unfolding action is usually investigated using a stop signal (or gostop) task. The data from the stop-signal task are often described using a horse-race model whose key assumption is that each process (i.e., go, stop) exhibits stochastic independence. Using three variations of a coincident timing task (i.e., go, gostop, and gostopgo) we extend previous considerations of stochastic independence by analysing the go latencies for prior effects of stopping. On random trials in the gostopgo task the signal sweep was paused for various times at various distances before the target. Significant increases in latency errors were reported on those trials on which the signal was paused (p <.005). Further analyses of the pause trials revealed significant effects for both the stopping interval (p <.001) and the pause interval (p <.05). Tukey post hoc analyses demonstrated increased latency errors as a linear function of the stopping interval, as expected, and decreased latency errors as a nonlinear function of the pause interval. These latter results indicate that the latencies of the go process, as reflected in the latency errors, may not exhibit stochastic independence under certain conditions. Various control mechanisms were considered in an attempt to explain these data. PMID- 12745833 TI - The relative efficacy of different forms of knowledge of results for the learning of a new relative timing pattern. AB - The goal of the present study was to determine the relative efficacy of verbal and auditory knowledge of results for promoting learning of a new constrained relative timing pattern. In a series of four experiments we compared the efficiency of verbal knowledge of results to that of auditory knowledge of results. The results of all four experiments revealed that verbal knowledge of result is a very effective source of information to promote learning of a new imposed relative timing pattern. Auditory knowledge of results favoured learning of a new relative timing pattern in a very limited set of circumstances. In the present study, this was only the case when movement velocity remained constant from one segment of the task to the next and if it resulted in an unfamiliar temporal pattern. The results of all four experiments also provided evidence that movement parameterization and relative timing are independent processes that can be developed in parallel. PMID- 12745834 TI - Creativity and inductive reasoning: the relationship between divergent thinking and performance on Wasons 246 task. AB - This study was an investigation of the relationship between potential creativityas measured by fluency scores on the Alternate Uses Testand performance on Wasons 246 task. As hypothesized, participants who were successful in discovering the rule had significantly higher fluency scores. Successful participants also generated higher frequencies of confirmatory and disconfirmatory hypotheses, but a multiple regression analysis using the stepwise method revealed that the frequency of generating disconfirmatory hypotheses and fluency scores were the only two significant factors in task outcome. The results also supported earlier studies where disconfirmation was shown to play a more important role in the later stages of hypothesis testing. This was especially true of successful participants, who employed a higher frequency of disconfirmatory hypotheses after receiving feedback on the first announcement. These results imply that successful participants benefited from the provision of feedback on the first announcement by switching to a more successful strategy in the hypothesis-testing sequence. PMID- 12745835 TI - Retrieval independence in successive recognition tasks. AB - In three experiments participants studied AB word pairs and completed two recognition tests. In the first recognition test, which was included in all three experiments, the B word had to be discriminated from two distractors that did not appear on the study list. In Experiment 1, in the second recognition test, an AB target was compared with distractors composed of words not on the study list. In Experiment 2, in the second recognition test, an AB target had to be discriminated from two other pairs that were created by randomly re-pairing A and B words that appeared on the study list. In Experiment 3, on the second recognition test, words from the study list were systematically re-paired to form distractors that contained either the same A term or the same B term as the target pair. Recognition of the B word on the first test was always at least partly independent of recognition of the AB pair on the second test. Even when recognition judgements were restricted to those for which the participants were most confident, all experiments demonstrated significant retrieval independence between the two tests. PMID- 12745836 TI - The influence of response--effect compatibility in a serial reaction time task. AB - Participants performed a serial reaction time task, responding to either asterisks presented at varying screen locations or centrally presented letters. Stimulus presentation followed a fixed second-order conditional sequence. Each keypress in the experimental groups produced a contingent, key-specific tone effect. The critical variation concerned the mapping of tones to keys. In Experiment 1, keypresses in one control condition produced noncontingent tone effects, while in another control condition there were no tone effects. In Experiment 2, three different keytone mappings were compared to a control condition without tone effects. The results show that tone effects improve serial learning when they are mapped to the response keys contingently and in a highly compatible manner. The results are discussed with reference to an ideomotor mechanism of motor sequence acquisition. PMID- 12745837 TI - Age of acquisition, ageing, and verb production: normative and experimental data. AB - Young and old adults were shown pictured or written verbs and asked to name them as quickly as possible. Simultaneous multiple regression was used to investigate which of a set of potential variables predicted naming speed. Age of acquisition was found to be an important predictor of naming speed in both young and old adults, and for both word and picture naming. Word frequency predicted picture naming speed only in older adults and failed to make any significant contribution to word-naming speeds for either group of participants. The respective loci and roles of age of acquisition and frequency in lexical processing are discussed in the light of these findings. PMID- 12745838 TI - Variable foreperiods and temporal discrimination. AB - Temporal judgements are often accounted for by a single-clock hypothesis. The output of such a clock is reported to depend on the allocation of attention. In the present series of experiments, the influence of attention on temporal information processing is investigated by systematic variations of the period preceding brief empty intervals to be judged. Two indicators of timing performance, temporal sensitivity, reflecting discrimination performance, and perceived duration served as dependent variables. Foreperiods ranged from 0.3 to 0.6 s in Experiments 1 to 4. When the foreperiod varied randomly from trial to trial, perceived duration was longer with increasing length of foreperiod (Experiments 1 and 3 with brief auditory markers and Experiment 4 with brief visual markers), an effect that disappeared with no trial-to-trial variations (Experiment 2). Longer foreperiods also enhanced performance on temporal discrimination of auditory empty intervals with a base duration of 100 ms (Experiments 1 and 5), whereas discrimination performance was unaffected for auditory intervals with a base duration of 500 ms (Experiment 3). The variable foreperiod effect on perceived duration also held when foreperiods ranged from 0.6 to 1.5 s (Experiments 57). Findings suggest that foreperiods appear to effectively modulate attention mechanisms necessary for temporal information processing. However, alternative explanations such as assimilation or compatibility effects cannot be totally discarded. PMID- 12745840 TI - Sequential effects in the lexical decision task: the role of the item frequency of the previous trial. AB - Two lexical decision experiments were conducted to determine whether there is a specific, localized influence of the item frequency of consecutive trials (i.e., first-order sequential effects) when the trials are not related to each other. Both low-frequency words and nonwords were influenced by the frequency of the precursor word (Experiment 1). In contrast, high-frequency words showed little sensitivity to the frequency of the precursor word (Experiment 2), although they showed longer reaction times for word trials preceded by a nonword trial. The presence of sequential effects in the lexical decision task suggests that participants shift their response criteria on a trial-by-trial basis. PMID- 12745841 TI - Homophone interference effects in visual word recognition. AB - In three lexical decision experiments and one progressive demasking experiment, performance on low-frequency heterographic homophones having a high-frequency mate was compared with performance on non-homophone target words with or without high-frequency orthographic neighbours. Robust homophone interference effects were observed in all experiments, as well as inhibitory effects of neighbourhood frequency. When speed-accuracy trade-offs were reduced, the homophone interference effects were found to be additive with effects of high-frequency orthographic neighbours. Furthermore, the size of homophone interference effects increased when pseudohomophone stimuli were presented among the nonwords. These results are tentatively interpreted within the framework of a bi-modal interactive activation model. PMID- 12745842 TI - Attending to the distractor and old/new discriminations in negative priming. AB - When participants ignore an irrelevant distractor they typically show impaired responding to that item if it becomes the relevant stimulus on a subsequent trial. In Experiment 1 (N = 64), a masked white colour name was presented briefly before a Stroop display. Negative priming in colour naming occurred when the colour of the lettering for the Stroop stimulus matched the colour name displayed in the first display, consistent with the proposal of temporal discrimination theory that negative priming arises because a recurrence of an unattended stimulus cannot readily be classified as old or new. Experiment 2 (N = 32) replicated negative priming in the interleaved-word display where participants had to name the red word from a pair of red and green words. In Experiment 3 (N = 32) and Experiment 4 (N = 28) the participants were required to attend to but not respond to the words in the prime display and name one of two interleaved words in the probe display. Negative priming was observed in this arrangement, consistent with the episodic retrieval theory of negative priming. The temporal discrimination model may need to be extended to situations in which the attended stimuli have different responses attached to them. PMID- 12745843 TI - How does orthographic knowledge influence performance on phonological awareness tasks? AB - Three experiments explored the nature of orthographic influences on performance on phonological awareness tasks. Experiment 1 demonstrated that adults find it easier to perform phoneme deletions on items where there is a direct correspondence between letters and target sounds (e.g., take the /r[se text]/ from struggle) than where there is not (e.g., take the /w[see text]/ from squabble). Analogous results were found in a phoneme reversal task. Spelling production ability tended to correlate more strongly with performance on the former type of item than on the latter, suggesting that elevated performance on phonological awareness tasks is associated with the use of orthographic information. Experiment 2 produced similar results in Grade 5 children. Experiment 3 suggested that adults cannot inhibit orthographic activation when it is disadvantageous to them, as they performed no better on items such as squabble when they were presented in pure blocks than when they were presented in mixed blocks. It is concluded that there are substantial automatic orthographic influences on phonological awareness task performance that need to be taken into account in interpreting data concerning the relationship between phonological awareness and reading. PMID- 12745844 TI - Individual stopping times and cognitive control: converging evidence for the stop signal task from a continuous tracking paradigm. AB - The present study introduces a continuous tracking procedure to investigate cognitive stopping in individual trials. Our measure of stopping performance had a mean similar to mean stopping times estimated in the stop signal paradigm, suggesting a common underlying process. Additional findings indicate that stopping performance and tracking performance were dissociable. First, while stopping times were primarily affected by stop signal modality, tracking performance was primarily affected by tracking difficulty. Second, tracking performance influenced tracking but not stopping in immediately following trials. Stopping influenced neither tracking performance nor stopping in immediately following trials. Finally, there was no correlation between tracking performance and stopping performance, or any dependency between them as found in the conditional means. PMID- 12745845 TI - Priming the rules of spelling. AB - This paper reports three spelling experiments that examined the effect of lexical priming through intervening items. In the first and second experiments, a strong effect of word priming on nonword spelling was found, even when two intervening filler items separated primetarget pairs. In addition, the absolute size of the effect was similar when one intervening item separated primetarget pairs and when two intervening items separated primetarget pairs. A much larger effect was found when no intervening items were used, however. This effect did not appear to be related to filler type, as Experiment 1 used nonword fillers, and Experiment 2 used word fillers. The third experiment examined the same effect with two intervening filler items, but instead used nonwords as primes (and thus examined a subsyllabic repetition effect). A similar-sized effect as that of the first and second experiments was found. The most plausible explanation of these results, which is consistent with the interactive dual-route model of spelling, is that they reflect the priming of soundspelling rules that people use to spell nonwords. PMID- 12745846 TI - Working-memory and auditory localization: demand for central resources impairs performance. AB - Four experiments explored possible roles for working memory in sound localization. In each experiment, the angular error of localization was assessed when performed alone, or concurrently with a working-memory task. The role of the phonological slave systems in auditory localization was ruled out by Experiments 1 and 2, while an engagement of central resources was suggested by the results of Experiment 3. Experiment 4 examined the involvement of visuo-spatial systems in auditory localization and revealed impairment of localization by the concurrent spatial working-memory task. A comparison of dual-task decrement across all four studies suggests that localization places greater demand on central than on spatial resources. PMID- 12745847 TI - The learning of goal-directed locomotion: a perception-action perspective. AB - This study was designed to better understand the process underlying the learning of goal-directed locomotion. Subjects walked on a treadmill in a virtual reality setting and were asked to cross pairs of oscillating doors. The subjects behaviour was examined at the beginning of the learning process (pretest), after 350 trials (intermediate test), and after 700 trials (posttest). The data were analysed at three different levels, each representing a specific aspect of the global response: performance outcome, displacement kinematics, and current arrival condition. While some aspects of performance outcome suggested the presence of a ceiling effect in the intermediate test, both displacement kinematics and current arrival condition clearly highlighted continuous transformations of the control mechanism involved. The learning process is best described as (1) the establishing of a relationship between specific information and a movement parameter and (2) the optimization of this relationship. The optimization process is characterized by the further exploration of the available behavioural repertoire and by the refinement of the dialogue between information and movement. PMID- 12745849 TI - Calcitriol shows greater persistence of treatment effect than betamethasone dipropionate in topical psoriasis therapy. AB - OBJECTIVE: To establish the efficacy and duration of remission post-treatment of calcitriol 3 micro g/g ointment in comparison with betamethasone dipropionate 0.05% ointment. METHODS: A randomized, multicentre trial was conducted in 258 adult patients with chronic plaque psoriasis. Calcitriol 3 microg/g ointment or betamethasone dipropionate 0.05% ointment was applied twice daily for 6 weeks or until complete clearance of lesions. Patients whose psoriasis cleared or were significantly improved and did not require treatment continuation at treatment endpoint were contacted over the following 8 weeks to determine whether relapse had occurred. RESULTS: Both treatments were efficacious; improvement in psoriasis or clearance of lesions (residual erythema was allowed) was recorded for 79% and 82% of patients receiving calcitriol or betamethasone dipropionate, respectively. Global improvement and global severity scores at treatment endpoint showed statistically significant differences in favour of betamethasone dipropionate (p<0.05); however, the absolute reduction in mean PASI (psoriasis area and severity index) was comparable between the groups. A statistically significantly (p<0.01) higher proportion of responders remained in remission (no worsening of the disease warranting new treatment) following calcitriol therapy (48%) than betamethasone therapy (25%). This is of potential importance to patients, physicians and healthcare suppliers. CONCLUSION: Twice-daily applications of either calcitriol 3 microg/g ointment or betamethasone dipropionate 0.05% ointment can be used to good effect in the treatment of chronic plaque psoriasis. However, the beneficial effect is likely to persist for longer following calcitriol treatment. PMID- 12745850 TI - Calcium homeostasis remains unaffected after 12 weeks' therapy with calcitriol 3 microg/g ointment; no correlation with extent of psoriasis. AB - OBJECTIVE: To assess the systemic safety of calcitriol 3 microg/g ointment (Silkis) ointment) in relation to body surface area (BSA) affected by chronic plaque psoriasis. METHODS: In this open-label, multicentre study, patients were divided into three parallel groups: 5% to <15% (n=23), 15% to <25% (n=18), 25% to 35% (n=18) based on BSA involvement. Ointment was applied topically twice daily for 12 weeks; patients were followed up for a further 8 weeks. RESULTS: There was no alteration of calcium homeostasis: the mean values of albumin-adjusted serum total calcium, as well as 24-hour urinary calcium and serum calcitriol levels, remained within normal ranges throughout treatment. No changes in calcium or phosphate homeostasis related to the area of psoriasis being treated with calcitriol ointment were detected. CONCLUSIONS: Reductions in the global severity score and BSA involvement, as well as results of the assessment of improvement, attested to the clinical efficacy of calcitriol 3 microg/g ointment in psoriasis. The study confirms the systemic safety of calcitriol 3 microg/g in psoriatic patients with 5-35% BSA involvement. PMID- 12745851 TI - The effect of PUVA treatment on sister chromatid exchange (SCE) values in psoriasis vulgaris patients. AB - BACKGROUND: Psoralen+ultraviolet A (UVA) (PUVA) is used successfully in the treatment of several skin diseases including psoriasis. PUVA has been reported to cause skin cancers, especially when used in high doses. In vivo and in vitro effects of different mutagens and carcinogens on DNA may be detected by sister chromatid exchange (SCE). OBJECTIVE: To investigate the mutagenic effects of different PUVA doses on DNA with SCE analysis. MATERIALS AND METHODS: Forty-two psoriasis patients under PUVA treatment were included in the study as the study group. The control group consisted of 22 psoriasis patients who did not receive PUVA treatment. The study group was divided into three groups according to PUVA doses. SCE/cell values were compared in the study and control groups and in the three dose-dependent groups of the study group. RESULTS: Mean SCE/cell values of the three dose-dependent patient groups were significantly higher than the control group (p<0.001), whereas there was not a statistically significant difference in themselves. CONCLUSION: PUVA treatment seemed to increase SCE values; however, there was not a correlation between PUVA doses and SCE frequencies. PMID- 12745852 TI - Treatment of cutaneous leishmaniasis with ketoconazole cream. AB - BACKGROUND: Cutaneous leishmaniasis (CL) is endemic in many countries. It has been recognized as a major public health problem in Iran. Many drugs have been suggested for the treatment of CL but most of them used intramuscularly or intravenously. Recently, a ketoconazole tablet has been used for the treatment of CL. OBJECTIVE: To compare the efficacy of ketoconazole cream with placebo in the treatment of CL. METHODS: The study was a double-blind, placebo-controlled trial. Patients with proven CL were treated with ketoconazole cream or placebo cream. The duration of treatment was 21 days and the patients were visited on days 7, 14 and 21. Follow-up was for 1 month after the end of treatment. RESULTS: The study was completed in 73 patients (45 male, 28 female). The mean age of the patients was 19.9 years. A total of 38 patients received ketoconazole and 35 patients were treated with the placebo cream. At the end of treatment (day 21), complete healing had occurred in 15.7% of the ketoconazole group in comparison with 14.3% of the patients who were treated with the placebo cream (p=0.86). One month after the end of treatment the lesions in 28.9% of the ketoconazole group and 22.8% of the placebo group were healed (p=0.55). CONCLUSION: Although the ketoconazole tablet has been reported to be effective in the treatment of some cases of CL, the low response rate in patients receiving ketoconazole cream indicates that it cannot be used as the single agent in the treatment of CL patients. PMID- 12745853 TI - Itraconazole versus terbinafine in the management of onychomycosis: an overview. AB - Ever since the introduction of itraconazole and terbinafine in the management of onychomycosis, there has been a revival of interest in the latter. In order to comprehend the intricate emerging scenario, an endeavor has been made to form a distinct outline in the shape of an overview on several of their facets. The review, therefore, envisages forming and facilitating instant decision-making. PMID- 12745854 TI - Acne in Apert's syndrome: treatment with isotretinoin. AB - Apert's syndrome is an uncommon disease characterized by synostosis of extremities, vertebrae and skull. A clear association between Apert's syndrome and acne vulgaris with resistance to usual acne treatments has been described. A case of Apert's syndrome treated with oral isotretinoin with good results is reported and the pathogenic mechanisms of acne in this syndrome are discussed. PMID- 12745855 TI - Topical nitrogen mustard ointment with occlusion for Langerhans' cell histiocytosis of the scalp. AB - BACKGROUND: Topical nitrogen mustard solution has been used as an effective alternative to corticosteroids for the treatment of cutaneous eruptions of Langerhans' cell histiocytosis (LCH). When used as an ointment under occlusion, nitrogen mustard may still be effective and possess less risk of unwanted side effects. METHODS: A patient with scalp LCH was treated topically with nitrogen mustard ointment 0.01% under occlusion. RESULTS: The lesions cleared in 3 weeks without irritation. CONCLUSION: Topical nitrogen mustard ointment 0.01% under occlusion is a well-tolerated, non-irritating treatment for scalp LCH. PMID- 12745856 TI - Severe resistant subacute prurigo successfully controlled by long-term cyclosporin. AB - The case of a 50-year-old woman with an intensely pruritic subacute prurigo resistant to multiple therapeutic regimens is described. Therapy with cyclosporin was started, achieving complete control of the disease, demonstrating a good safety profile during long-term treatment. This is the first report on the successful use of cyclosporin for subacute prurigo. PMID- 12745857 TI - Cutaneous anthrax associated with facial palsy: case report and literature review. AB - BACKGROUND: Anthrax is primarily an animal disease. Bacillus anthracis, the causal agent in anthrax, is a Gram-positive rod. Humans can acquire anthrax by industrial exposure to infected animals or animal products. METHODS: Reported here is the case of a 48-year-old male farm worker from Iran with a history of direct contact with herds. He presented after 6 days of fever with toxicity and a crusted ulcer on the face that was later confirmed bacteriologically to be cutaneous anthrax. He was treated with large doses of intravenous penicillin and corticosteroids along with multiple subcutaneous epinephrine injections that were used to control the infection and massive facial edema. RESULTS: After 14 days, he partially recovered; however, ipsilateral facial nerve palsy developed and persisted despite therapeutic efforts. CONCLUSION: It is not possible to conclude whether early diagnosis and treatment of anthrax results in a lower risk of complications. Facial palsy can be added to the list of variable complications of the cutaneous effects of anthrax. PMID- 12745858 TI - A case of recurrent acute generalized exanthematous pustulosis due to beta-lactam antibiotics: a case report. AB - A case of acute generalized exanthematous pustulosis (AGEP) is presented. The case is notable for the recurrent episodes of AGEP, caused by three beta-lactam antibiotics (piperacillin, ceftazidime, and meropenem) in septicemic patient. The case represents the first report of the reaction developing in response to these three antibiotics. The report is also notable for the spontaneous resolution of the rash in all the three episodes. PMID- 12745859 TI - Effective treatment of relapsing idiopathic nodular panniculitis (Pfeifer-Weber Christian disease) with mycophenolate mofetil. AB - Relapsing idiopathic nodular panniculitis is the term used to describe a group of diseases that presents as subcutaneous inflammatory nodules, fever and systemic symptoms and histopathologically displays inflammation within the fat lobules. There is no specific test for diagnosis and extensive investigations are required to exclude systemic causes of panniculitis. No uniform effective therapy is available and various drugs used include mainly corticosteroids alone or in combination with other immunosuppressive agents. Presented in this paper is an intractable case of idiopathic nodular panniculitis whose corticotherapy failed and could not be continued because of serious adverse effects. The rapid and good therapeutic response of the patient to mycophenolate mofetil monotherapy is discussed. PMID- 12745861 TI - False-negative myocardial scintigraphy in balanced three-vessel disease, revealed by coronary pressure measurement. AB - In nuclear perfusion imaging of the myocardium, a false-negative test result in patients with balanced three-vessel disease is a well-known pitfall. This paper describes a patient with typical chest pain and a negative myocardial perfusion scintigram. At coronary angiography, intermediate stenoses in the left anterior descending (LAD), left circumflex (LCX), and right coronary (RCA) arteries were present. Fractional flow reserve, measured by coronary pressure measurement, was 0.54, 0.56, and 0.66 respectively for the LAD, LCX, and RCA, unequivocally demonstrating the presence of balanced three-vessel disease. The patient underwent successful bypass surgery and remained event-free thereafter. PMID- 12745860 TI - Use of intravenous abciximab as adjunctive therapy for carotid angioplasty and stent placement. AB - The benefit of intravenous abciximab as an adjunctive to percutaneous coronary intervention has been demonstrated in large-scale randomized studies. The role of intravenous abciximab is being defined in carotid angioplasty and stent placement as the procedure is gaining popularity for the treatment of high-grade carotid stenosis in patients considered high-risk for carotid endarterectomy. This paper summarizes the pathophysiological basis and the available data for the use of abciximab as an adjunct to carotid artery stenting. PMID- 12745863 TI - A tortuous distal carotid artery: how to overcome the problem, with the aim of guaranteeing distal protection. AB - This report describes a case of stent implantation in a right internal carotid artery with severe distal tortuosity, which caused difficulty in advancing a filter protection device. Advancement of the introducing sheath at the level of the carotid bifurcation and exchange for a third-generation distal filter protection device overcame the problem. PMID- 12745862 TI - Safe and effective direct implantation of a new stent through 5 F. guiding catheters with delivery from the radial artery: initial results of a prospective registry. AB - OBJECTIVES: To evaluate the safety and efficacy of the direct implantation of a new stent via the radial artery through a 5 F. guiding catheter. Background advances in the design of stents and stent delivery systems have facilitated the performance of direct stenting and the use of thinner guiding catheters. METHODS: This registry enrolled prospectively 125 patients (147 lesions, 20.4% AHA/ACC class B2/C) who underwent elective percutaneous coronary revascularization procedures for stable or unstable angina between November 2000 and March 2001. RESULTS: Cannulation of the radial artery was attempted in 92.7% and was successful in 91.0% of cases. Direct stenting was successful in 88.7% of lesions and procedural success was 99.3%. In-hospital major adverse cardiac events occurred in 1.6% of cases (one death, one semi-urgent coronary artery bypass operation). The final rate of successful stent implantation through 5 F. guiding catheters was 96.7%. There were no access-site-related complications. Failure to cross the lesion occurred in 10% of attempts. At a mean follow-up of 7 +/- 2.8 months after discharge from hospital, 79% of patients had remained free of angina, and 89% had remained free of ischemic events. CONCLUSIONS: Direct stenting with a new stent design was safe, effective, and could be accomplished through 5 F. guiding catheters with favorable long-term clinical outcomes. PMID- 12745864 TI - The significance of subclavian artery injection prior to surgical myocardial revascularization. AB - Successful arterial revascularization using the internal thoracic artery is dependent on unobstructed inflow through the subclavian artery. Systematic physical examination should discover subclavian stenosis; however, simple routine injection into the orifice of the subclavian artery during the diagnostic catheterization may avoid a catastrophic outcome. PMID- 12745865 TI - Distal embolization at the site of a stenotic distal anastomosis of a saphenous vein graft, leading to dye entrapment. AB - This case report describes the entrapment of contrast media after recanalization of a recently occluded saphenous vein graft with balloon predilation, thrombectomy and stent implantation. Recanalization of the respective coronary artery was performed, and the entrapped contrast media within the saphenous vein graft progressed to the left circumflex artery. PMID- 12745866 TI - Treatment of recurrent coronary rupture by implantation of three coronary stent grafts. AB - The authors report a case of recurrent left anterior descending artery rupture during coronary interventions in a 70-year-old man. Coronary artery rupture was treated successfully by percutaneous coronary stent-graft implantation. Based on this experience, the authors advise against repeat angioplasty of a coronary artery which has ruptured during a prior intervention. Membrane-covered stents should be the first choice in the treatment of life-threatening coronary artery rupture. PMID- 12745867 TI - Prolonged systemic delivery of tirofiban in a thrombus-laden saphenous vein graft. AB - Results from previous trials have shown conflicting results from local delivery of thrombolytic agents to diminish thrombus burden before intervention in native coronary arteries and saphenous vein grafts. We described a patient with an acute coronary syndrome who was treated for 24 hours with systemic tirofiban (Aggrastat), a glycoprotein IIb/IIIa inhibitor, for the treatment of a degenerated saphenous vein graft with a TIMI grade 4 thrombus (large-sized thrombus). Angiographic evaluation 48 hours later revealed complete resolution of the thrombus with normal coronary blood flow. PMID- 12745868 TI - Acute two-vessel coronary closure in a patient with Down's syndrome. AB - Myocardial infarctions are rare in patients with Down's syndrome. This paper reports an unusually aggressive presentation of two-vessel simultaneous coronary occlusion during an intended percutaneous intervention. Since survival in patients with Down's syndrome is improving, encounters with late (and perhaps unusual) sequelae of coronary artery disease are expected to increase. PMID- 12745869 TI - The YUMIKO catheter: a useful tool for angiography of the right internal mammary artery via a right brachial approach. AB - The YUMIKO catheter (Goodman, Nagoya, Japan) was recently developed for a left internal mammary artery (IMA) angiography with a right radial or brachial approach. The present authors experienced an interesting case where the YUMIKO catheter was useful for a right IMA angiography via a right brachial artery. A 53 year-old man with bilateral IMA grafts underwent follow-up coronary angiography via a right brachial artery. Native coronary artery and left IMA angiography were performed without difficulty using the Judkins Right and Left and YUMIKO catheters. Angiography of the right IMA was attempted with the Judkins Right catheter and IMA catheter, resulting in a nonselective angiogram with poor imaging. The YUMIKO catheter, however, enabled smooth cannulation to the right IMA and provided good images of the selective right IMA angiography. PMID- 12745870 TI - Determination of the human cytochrome P450s involved in the metabolism of 2n propylquinoline. AB - 1 2n-Propylquinoline (2nPQ) is a newly developed drug for visceral antileishmaniasis and its activity has been previously evaluated in mice following oral administration. The study was carried out to investigate the kinetic formation of 2nPQ metabolites and to characterize the human liver CYP forms involved in its oxidative metabolism. 2. The inhibition of 2nPQ metabolite formation by specific substrates or inhibitors of CYP forms and correlation studies were performed in human liver microsomes. 2nPQ biotransformation was then studied in human lymphoblasts expressing specific CYPs and microsomal epoxide hydrolase. 3. Three major metabolites were produced by human liver microsomes and their structures were identified by ESI-LC/MS: dihydroxy-2n-propylquinoline, 3' hydroxy-2n-propylquinoline and 1'-hydroxy-2n-propylquinoline. An intermediary metabolite, epoxy-2n-propylquinoline, formed by CYP was also biotransformed by microsomal epoxide hydrolase into dihydroxy-2n-propylquinoline. 4. 2nPQ oxidation follows Michaelis-Menten kinetics. In human liver microsomes, its metabolism was extremely inhibited by pilocarpine, coumarin and diethyldithiocarbamate. From a panel of 12 human liver microsome samples, the rate of 2nPQ oxidation was highly correlated with the activities of CYP2A6 and CYP2E1. Human lymphoblasts expressing specific CYPs showed the involvement of CYP2A6, CYP2E1 and CYP2C19. 5. The results indicate that 2nPQ metabolites are 3'- and 1'-hydroxylated by human liver microsomes and an epoxy-2n-propylquinoline is biotransformed into a dihydroxy-2n-propylquinoline by microsomal epoxide hydrolase. PMID- 12745871 TI - Curcumin is a potent inhibitor of phenol sulfotransferase (SULT1A1) in human liver and extrahepatic tissues. AB - 1. Curcumin has anti-carcinogen effects and is under clinical evaluation as a potential colon cancer chemopreventive agent. The first aim was to see whether curcumin inhibited phenol sulfotransferase (SULT1A1) and, if so, to study the variability of the IC(50) of curcumin for SULT1A1 in 50 human liver samples. For comparative purposes, the inhibition of catechol sulfotransferase (SULT1A3) in five human liver specimens was studied. The second aim was to measure the IC(50) of curcumin against SULT1A1 in five samples of human duodenum, colon, kidney and lung. 2. Curcumin was a potent inhibitor of SULT1A1 in human liver; the mean +/- SD and median of IC(50) were 14.1 +/- 7.3 nM and 12.8 nM, respectively. The IC(50) ranged from 6.2 to 30.6 nM between the 5th and 95th percentiles and the fold of variation was 4.9. The distribution of IC(50) was positively skewed (skewness 1.2) and deviated from normality (p = 0.0004). 3. Curcumin inhibited human SULT1A3, and the inhibition was studied in five liver specimens with an IC(50) of 4324 +/- 1026 nM. This inhibition was greater than the IC(50) of curcumin for SULT1A1 (p < 0.0001). 4. In the extrahepatic tissues, the IC(50) of curcumin for SULT1A1 was 25.9 +/- 4.8 nM (duodenum), 25.4 +/- 6.8 nM (colon), 23.4 +/- 2.2 nM (kidney) and 25.6 +/- 5.6 nM (lung). Inhibition in these tissues is greater than that of curcumin for SULT1A1 in human liver (p < 0.0001). 5. In conclusion, curcumin is a potent inhibitor of SULT1A1 in human liver, duodenum, colon, kidney and lung. The IC(50) of curcumin for SULT1A1 varied 4.9-fold in human liver. The comparison of the present data with those of the literature revealed that the IC(50) of curcumin in the liver and extrahepatic tissues is one order of magnitude lower that the peak serum concentration of curcumin after therapeutic doses of 4 g to humans. PMID- 12745872 TI - Differential metabolism of midazolam in mouse liver and intestine microsomes: a comparison of cytochrome P450 activity and expression. AB - 1. Although multiple cytochrome P450s (CYP) contribute to hepatic phase I metabolism, CYP3A is the principal subfamily present in human and mouse small intestine. 2. Differences in phase I metabolism were investigated using midazolam (MDZ) hydroxylation in mouse liver and intestinal microsomes. The net MDZ metabolite formation rate in intestinal microsomes was approximately 30% that of liver microsomes (at 250 micro M MDZ). 3. Quantitative Western blotting with anti CYP3A1 antibody detected two bands of immunoreactive protein in both liver and intestinal samples, 2.24 +/- 0.27 (mean +/- SD, n = 3) and 0.64 +/- 0.08 pmol mg( 1) protein, respectively. Qualitative Western blotting with anti-CYP2C11 antibody detected a single band of immunoreactive protein in liver microsomes and no signal in intestinal samples (1 micro g sample). 4. Ketoconazole potently inhibited formation of both alpha- and 4-OH-MDZ metabolites in intestinal microsomes (IC(50)' of 0.126 +/- 0.010 and 0.0955 +/- 0.014 micro M, respectively) and of 4-OH-MDZ formation in mouse liver microsomes (IC(50) of 0.041 +/- 0.003 micro M). However, ketoconazole (5 micro M) did not produce 50% inhibition of alpha-OH-MDZ formation in mouse liver microsomes. Inhibition by ritonavir (5 micro M) produced similar results. 5. MDZ hydroxylation is predominately CYP3A dependent in mouse intestine (compared with mouse liver) since CYP2C is not expressed in the intestine. The importance of CYP3A in the mouse intestine appears to mirror that in humans. PMID- 12745873 TI - Uptake of rosuvastatin by isolated rat hepatocytes: comparison with pravastatin. AB - 1. The liver is the target organ for the lipid-regulating effect of rosuvastatin, a new 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, and liver selective uptake of this drug is therefore a desirable property. The uptake kinetics of rosuvastatin were investigated and compared with those of pravastatin using isolated rat hepatocytes. 2. Uptake for both drugs involved both active transport and passive diffusion processes. The Michaelis constant (K(m)) of uptake rate for rosuvastatin (9.17 micro M) was approximately half that for pravastatin (16.5 micro M). However, the maximum uptake rate (V(max)) and carrier mediated uptake clearance (V(max)/K(m)) of rosuvastatin were significantly (p < 0.01) greater than those of pravastatin, and a larger contribution of carrier mediated uptake clearance to total uptake clearance was shown for rosuvastatin (contribution ratio 0.903 versus pravastatin 0.654). 3. Sodium and chloride ions did not play a significant role in the uptake of rosuvastatin and pravastatin, but the uptake of both drugs was inhibited both by depletion of cellular ATP and by organic anions such as bromosulfophthalein. 4. Rosuvastatin competitively inhibited the uptake of pravastatin, with an inhibition constant (K(i)) (2.75 micro M) relatively similar to its K(m). 5. The results suggest that an organic anion transport protein is the main mediator of the hepatic uptake of rosuvastatin and pravastatin, which occurs in an ATP-dependent manner. Our results indicated that rosuvastatin was taken up by the hepatocytes via the same transport systems as pravastatin, but with a greater affinity and efficiency than pravastatin. PMID- 12745874 TI - Glucose inhibition of the induction of CYP2E1 mRNA expression by ethanol in FGC-4 cells. AB - 1. Rats fed intragastrically with ethanol-containing diets made with low levels of carbohydrates have greater CYP2E1 induction than rats fed similar diets made with high carbohydrate levels. 2. FGC-4 rat hepatoma cells were used to test the hypothesis that carbohydrates could down-regulate ethanol-induced CYP2E1 induction. 3. FGC-4 cells grown in a glucose-free media and treated with 1-100 mM ethanol for 24 h exhibited a dose-dependent increase (p < 0.05) in CYP2E1, with maximum mRNA steady-state (3.8-fold) or protein (3.1-fold) levels measured at 30 or 100 mM ethanol, respectively. 4. In cells treated with 30 mM ethanol, a glucose concentration-dependent inhibition (p < 0.05) of CYP2E1 mRNA was observed between 2.5 and 10 mM glucose. 5. Induction by 30 mM ethanol of CYP2E1 protein was reduced in cells co-treated with 1 mM or greater glucose concentration and complete inhibition was measured with 5 mM glucose co-treatment. 6. These data demonstrate that under culture conditions of extremely low carbohydrate concentrations: (1) ethanol treatment of FGC-4 cells results in elevated steady state levels of CYP2E1 mRNA and protein; and (2) glucose inhibits this increase. 7. It is concluded that glucose can negatively regulate CYP2E1 expression and could at least partially explain the greater induction of hepatic CYP2E1 in rats fed low carbohydrate ethanol-containing diets compared with high carbohydrate diets at the same ethanol level. PMID- 12745875 TI - In vivo and in vitro disposition profile of tezosentan, an intravenous dual endothelin receptor antagonist, in humans. AB - 1. The plasma concentration-time profile of the (14)C-labelled endothelin receptor antagonist tezosentan in healthy male volunteers after a 1-h infusion at 100 mg h(-1) followed a biphasic decline with half-lives of 3-5 min for the initial disposition phase and approximately 4 h for the terminal phase. 2. Tezosentan was predominantly excreted unchanged into faeces, whereas less than 5% of the dose was excreted as unchanged drug in urine. Two isomeric, hydroxylated metabolites (M1, M2) were detected in faeces representing 2-5% of the total radioactivity. 3. In vitro, with human liver microsomes and primary hepatocytes, tezosentan was metabolized at very low rates. Upon prolonged incubation with human hepatocytes for 24 h, formation of the hydroxylated metabolite M1 and a glucuronic acid conjugate, M3, was observed. 4. No relevant inhibition of the human cytochrome P450 (CYP) forms, CYP1A2, 2C9, 2C19, 2D6 and 3A4, was observed in vitro at tezosentan concentrations largely exceeding those observed in clinical trials. 5. In human blood, tezosentan was highly bound to plasma proteins, mainly albumin, and hardly penetrated into red blood cells. PMID- 12745877 TI - Hepatotoxicity of ketoconazole in Sprague-Dawley rats: glutathione depletion, flavin-containing monooxygenases-mediated bioactivation and hepatic covalent binding. AB - 1. This study has examined ketoconazole (KT)-induced hepatotoxicity in vivo and in vitro, using male Sprague-Dawley rats with [(3)H]KT (1.5 micro Ci mg(-1)) at 40 and 90 mg KT kg(-1) doses. Blood and liver samples were collected from 0 to 24 h for alanine aminotransaminase (ALT), glutathione (GSH) and covalent binding analyses. 2. Covalent binding occurred as early as 0.5 h, peaked at 2 h (0.026 +/ 0.01 nmol KT mg(-1) protein) and 8 h (0.088 +/- 0.04 nmol KT mg(-1) protein) for 40 and 90 mg KT kg(-1) doses, respectively. ALT levels increased at 0.5 h for the 40 and 90 mg KT kg(-1) doses (44.3 and 56.4 U ml(-1), respectively) relative to control, 22.7 U ml(-1). At 24 h, the 90 mg KT kg(-1) dose reduced hepatic GSH levels from 9.92 +/- 1.1 to 4.76 +/- 0.3 nmol GSH mg(-1) protein. 3. The role of the flavin-containing monooxygenases (FMO) utilized Sprague-Dawley microsomes with 1, 10 and 100 micro M [(3)H]KT. Maximum covalent binding occurring at 100 micro M KT. Heat inactivation of microsomal FMO significantly decreased covalent binding by 75%, whereas 1 mM GSH significantly reduced covalent binding by 65%. 4. Thus, KT-induced hepatotoxicity is dose- and time-dependent and appears to be FMO mediated, in part, to metabolites that may react with protein and, possibly, GSH. PMID- 12745876 TI - Pharmacokinetics, metabolism and excretion of the glycine antagonist GV150526A in rat and dog. AB - 1. The pharmacokinetics, metabolic fate and excretion of 3-[-2(phenylcarbamoyl) ethenyl-4,6-dichloroindole-2-carboxylic acid (GV150526), a novel glycine antagonist for stroke, in rat and dog following intravenous administration of [C14]-GV150526A were investigated. 2. Studies were also performed in bile duct cannulated animals to confirm the route of elimination and to obtain more information on metabolite identity. 3. Metabolites in plasma, urine and bile were identified by HPLC-MS/MS and NMR spectroscopy. 4. GV150526A was predominantly excreted in the faeces via the bile, with only trace metabolites of radioactivity in urine (< 5%). Radioactivity in rat bile was predominantly due to metabolites, whereas approximately 50% of the radioactivity in dog bile was due to parent GV150526. 5. The principal metabolites in bile were identified as glucuronide conjugates of the carboxylic acid, whereas in rat urine the main metabolite was a sulphate conjugate of an aromatic oxidation metabolite. Multiple glucuronide peaks were observed and identified as isomeric glucuronides and their anomers arising from acyl migration and muta-rotation. PMID- 12745878 TI - Effects of 1,2-dibromoethane on haematopoiesis in the chick embryo. AB - 1. Chick embryo in ovo was used to investigate the effects of 1,2-dibromoethane (DBE) on haematopoiesis at a developmental stage where the primitive erythroid cells divide and differentiate in circulation. 2. Early after DBE treatment on embryonic day 3, annexin V/propidium iodide labelling showed acute cell death of erythroid elements, which was subsequently compensated for by the release of immature cells into the circulation. Simultaneously, the comet assay indicated increased DNA damage in DBE-exposed blood cells when compared with controls. 3. After embryonic day 5, there was no indication for ongoing prominent cell death in the DBE-treated group. However, the DNA damage assessed by the comet assay persisted until embryonic day 10 in the peripheral blood cells, and for even longer in cells from thymus and bursa. 4. The kinetics of DNA fragmentation in both erythroid and lymphoid cells implied genotoxic damage by DBE to the stem cells of the definitive elements and transmission of this damage through the successive cell generations. 5. The early chick embryo provides a suitable alternative to mammalian models for investigation of long-term effects of xenobiotics on haematopoiesis. PMID- 12745879 TI - Porphyrin-mediated boron neutron capture therapy: evaluation of the reactions of skin and central nervous system. AB - PURPOSE: Recently, various boronated porphyrins have been shown to preferentially target a variety of tumour types. Of the different porphyrins evaluated, copper tetra-phenyl-carboranyl porphyrin (CuTCPH) is a strong candidate for future preclinical evaluation. In the present study, the responses of two critical normal tissues, skin and central nervous system (CNS), to boron neutron capture (BNC) irradiation in the presence of this porphyrin were evaluated. MATERIALS AND METHODS: Standard models for the skin and spinal cord of adult male Fischer 344 rats were used. CuTCPH was administered by intravenous infusion at a dose of 200 mg x kg(-1) body weight, over 48 h. The thermal beam at the Brookhaven Medical Research Reactor was used for the BNC irradiations. The 20-mm diameter irradiation field, for both the skin and the spinal cord, was located on the mid dorsal line of the neck. Dose-response data were fitted using probit analysis and the doses required to produce a 50% incidence rate of early and late skin changes or myeloparesis (ED(50) +/- SE) were calculated from these curves. RESULTS: Biodistribution studies indicated very low levels of boron (<3 microg x g(-1)) in the blood 3 days after the administration of CuTCPH. This was the time point selected for radiation exposure in the radiobiological studies. Levels of boron in the CNS were also low (2.8 +/- 0.6 microg x g(-1)) after 3 days. However, the concentration of boron in the skin was considerably higher at 22.7 +/- 2.6 microg x g(-1). Single radiation exposures were carried out using a thermal neutron beam. The impact of CuTCPH-mediated BNC irradiation on the normal skin and CNS at therapeutically effective exposure times was minimal. This was primarily due to the very low blood boron levels (from CuTCPH) at the time of irradiation. Analysis of the relevant dose-effect data gave compound biological effectiveness factors of about 1.8 for skin (moist desquamation) and about 4.4 for spinal cord (myeloparesis) for CuTCPH. These values were based on the BNC radiation doses to tissues calculated using the blood boron levels at the time of irradiation. CONCLUSIONS: CuTCPH-mediated BNC irradiation will not cause significant damage to skin and CNS at clinically relevant radiation doses provided that blood boron levels are low at the time of radiation exposure. PMID- 12745880 TI - Bronchoalveolar lavage and interstitial cells have different roles in radiation induced lung injury. AB - PURPOSE: To determine the contribution of intra-alveolar cells as opposed to cells fixed in the interstitium in the development of radiation-induced lung injury. MATERIALS AND METHODS: C3H/HeN mice were irradiated to the thorax with various doses of radiation. The cellular composition and cytokine production were assessed in the two sites by histological staining and RNase protection assay. RESULTS: Following thoracic irradiation, there was an initial decrease in the number of bronchial alveolar lavage (BAL) cells that was followed after 2 months by a dose-dependent increase up to 4 months. Foamy Mac-1 positive macrophages were present early in the BAL populations, which also expressed the pro inflammatory cytokines TNF-alpha, IL-1alpha and IL-1beta, but this response subsided by the time of onset of pneumonitis (3 months). In contrast, in whole lung tissue there was a steady increase in Mac-1 positive cells and increased expression of TNF-alpha, IL-1alpha and IL-1beta mRNAs to maximum levels at 3-4 months. CONCLUSIONS: These data indicate distinct temporal and spatial changes in pro-inflammatory cytokine gene expression in different cellular compartments of the irradiated lung. BAL cells became inflammatory early on, but interstitial cells became involved later and were probably more involved in contributing to the pneumonitis. PMID- 12745881 TI - Evaluation of risk factors for lung tumour induction in rats exposed to either NpO(2) or PuO(2) aerosols. AB - PURPOSE: To compare the incidence of each lung tumour type after inhalation exposure of rats to either NpO(2) or industrial PuO(2) aerosols, which have a similar size. MATERIALS AND METHODS: Male Sprague-Dawley rats were exposed once and followed during their whole life span. At the end of their life, the whole lungs were fixed, embedded and cut into thin sections for histological analysis. The presence of tumours was evaluated on three distinct levels of the lobes for phenotype determination to establish dose-effect relationships. RESULTS: In the range of lung doses studied (0.05 to more than 50 Gy), the general trend was an increased frequency of all types of tumours after inhalation exposure to neptunium compared with plutonium. The linearity of the lower part of the dose effect relationships for all malignant lung tumours leads to the conclusion that NpO(2) is 3.3-fold more carcinogenic than PuO(2). CONCLUSIONS: According to a linear extrapolation of the data on malignant lung tumour incidence collected among all studies reported on actinide oxide carcinogenesis, the risk of lung tumour appears to vary over a factor of about 10 depending on the nature and/or size of the aerosol. This variation has to be taken into account for a realistic assessment of tumour risk. PMID- 12745882 TI - Absorbed fraction of radon progeny in human bronchial airways with bifurcation geometry. AB - PURPOSE: The absorbed fraction, defined as the portion of the initial particle energy which is absorbed in the tissue of interest, was calculated, under bifurcation geometry of the airway tubes, for alpha-particles emitted from radon progeny in the human respiratory tract. The results are given for all branching generations and compared with the data obtained for the commonly used infinite straight cylinders adopted by the International Commission on Radiological Protection (ICRP) Report 66. MATERIALS AND METHODS: A model was created to calculate the absorbed fraction of alpha-particle energy in the human lung using bifurcation geometry. Monte Carlo simulations of alpha-particle propagation in tissue and air were performed. The stopping powers of alpha-particles were adopted from the International Commission of Radiation Units and Measurements (ICRU) Report 49. RESULTS: The absorbed fractions for the bifurcation geometry are given for the 15 generations in the tracheobronchial tree for alpha-particle energies of 6 and 7.69 MeV. The sources were assumed to be the fast and slow moving mucus. CONCLUSIONS: Comparisons with ICRP66 data reveal that the assumption of long, straight cylinders was appropriate in some cases, but not in all. Adoption of the absorbed fractions obtained from the bifurcation model instead of the ICRP66 data caused 'redistribution' of doses in the bronchial (BB) and bronchiolar (bb) regions. PMID- 12745883 TI - Effect of radiation on Ewing tumour subpopulations characterized on a single-cell level: intracellular cytokine, immunophenotypic, DNA and apoptotic profile. AB - PURPOSE: Adhesion molecules, cytokines and their corresponding cell-surface receptors are involved in intercellular signalling pathways, radioresistance and metastasis-mediating mechanisms of malignant cells. The aim was the characterization of changes in the marker profile of Ewing tumour cell subpopulations under the influence of radiation. MATERIALS AND METHODS: Three Ewing tumours were characterized in vitro and in vivo in a xenograft model before and after radiation by five-parameter flow cytometry. Antibodies directed against cell surface and intracellular antigens, apoptosis-associated markers and the DNA dye 7-aminoactinomycin D were used. RESULTS: Tumour cell subpopulations were identified by expression of adhesion molecules and cytokine receptors, intracellular cytokines, apoptotic markers and DNA content. Heterogeneous changes of flow cytometric profile were identified on tumour cell subpopulations after radiation. CONCLUSIONS: The changed profile of tumour cells under radiation might be associated with biological changes of tumour subpopulations in view of radioresistance and metastatic potential and might be useful to identify intercellular regulation mechanisms and to define parameters being predictive for a response to therapy. PMID- 12745884 TI - Variation in radiation-induced apoptosis in ataxia telangiectasia lymphoblastoid cell lines. AB - PURPOSE: To investigate and compare the ability of Epstein-Barr virus-transformed lymphoblastoid cell lines (LCL) from healthy individuals (normals) and ataxia telangiectasia (A-T) patients to undergo apoptosis after exposure to ionizing radiation. MATERIALS AND METHODS: Four normal and eight A-T LCL were exposed to doses of up to 20 Gy ionizing radiation. Apoptosis induction was studied 24 h after irradiation using three different methods: measurement of caspase-3 activity, PARP-1 cleavage and estimation of the sub-G(1) cell fraction. RESULTS: Of the eight A-T LCL tested, all harbouring truncating ATM mutations, five had a higher level of spontaneous apoptosis than the normal LCL as assessed by the sub G(1) cell fraction. Four of the eight A-T LCLs showed a similar level of radiation-induced apoptosis after exposure to 5 Gy as the normal LCL. The other four A-T LCL showed a greater radiation-induced apoptotic response, as assessed by at least one of the three techniques. CONCLUSIONS: LCL from A-T patients can undergo ionizing radiation-induced apoptosis in spite of a defect in ATM-p53 dependent signalling pathways. However, the apoptotic response is characterized by a large degree of variability between the A-T cell lines, the causes of which remain to be established. PMID- 12745885 TI - Radiation-induced chromosome breaks in ataxia-telangiectasia cells remain open. AB - PURPOSE: To determine if broken chromosome-end healing mechanisms through the addition of new telomeric sequences exist in cells having difficulties in rejoining the ends of broken chromosomes. MATERIALS AND METHODS: A full-colour painting protocol of all human chromosomes by FISH was combined with a telomeric and centromeric labelling using PNA probes to characterize the rejoining pattern and telomere status of radiation-induced chromosome breaks in ataxia telangiectasia (A-T) and normal lymphoblastoid cell lines. RESULTS: It was first established that the cell lines used for chromosome healing analysis were chromosomally stable. FISH analysis provided evidence that the frequency of deleted chromosomes, apparently unrejoined, was much higher in A-T than in normal cells, as expected by the role of ATM in cell-cycle control, as well as in DNA repair. In spite of their high frequency, broken chromosome ends in A-T cells do not seem to act as substrates for telomerase since additional terminal telomere sequences (more than the 92 expected pairs) indicative of chromosome healing were never observed. Broken chromosome ends in A-T cells remained open. CONCLUSION: The disability of cells to rejoin broken chromosome ends does not lead to the healing of DSBs by the acquisition of new telomeric sequences. PMID- 12745887 TI - Preparedness to respond to possible acts of nuclear terrorism: some strategies and recommendations. PMID- 12745886 TI - Differential effectiveness of solar UVB subcomponents in causing cell death, oncogenic transformation and micronucleus induction in human hybrid cells. AB - PURPOSE: (1). To determine the biological effectiveness of two solar ultraviolet (UVB) spectra with different lower wavelength thresholds for oncogenic transformation and micronucleus induction in CGL1 cells; (2). to investigate whether the action spectra for short- and long-term effects are similar; and (3). to investigate possible links between transformation and other delayed effects. MATERIALS AND METHODS: Two spectra were derived from a solar UV simulator by using two filters: the first transmitted radiation with lambda > 284 nm, the second with lambda > 293 nm. The resulting spectra have the same UVA, but different UVB components (lambda between 284 and 320 nm, 19 W m(-2), and lambda between 293 and 320 nm, 13 W m(-2)). CGL1 cells were irradiated with 466 J m(-2) with lambda > 284 nm and 1582 J m(-2) with lambda > 293 nm. These doses were approximately equilethal. The endpoints examined were oncogenic transformation, and centromere-positive and -negative micronucleus frequencies in the directly irradiated cells and in transtheir progeny. RESULTS: At equilethal doses, the oncogenic transformation frequency in the directly irradiated cells was greater by a factor of at least 7 for lambda > 284 nm irradiation compared with lambda > 293 nm. The micronucleus induction frequency was also significantly higher with the lambda > 284 spectrum. Consistent with our previous findings, no delayed micronucleus formation was found in the progeny of cells exposed to lambda > 293 nm, while a threefold elevation above controls was seen in the progeny of cells exposed to lambda > 284 nm irradiation. This was also the case for formation of micronuclei with a centromere. CONCLUSIONS: It was found that: (1). for equilethal doses the lambda > 284 nm spectrum was more biologically effective than the lambda > 293 nm spectrum for induction of oncogenic transformation and micronucleus formation; and (2). the higher effectiveness of the lambda > 284 nm spectrum found at equilethal doses for delayed effects in the progeny of irradiated cells resembles that found for transformation. The results suggest that the UVB action spectrum for cell killing is different from that of some delayed effects, and from that of transformation. PMID- 12745889 TI - Editorial: Basic research and clinical practice. PMID- 12745890 TI - Creating and being created: the changing panorama of paediatric rehabilitation. AB - Paediatric rehabilitation as a discipline is rapidly changing, especially during the last decades. In the past, paediatric rehabilitation was characterized by merely adult intervention strategies in a miniaturized form, delivered by a merely adult patients-oriented profession. Theories on childhood development, however, changed, as did the focus of interventions: from impairments to function, from the child itself to family, community and peers. The call for outcome-oriented and evidence-based medicine lastly, changed paediatric rehabilitation into a mature paediatric profession with it's own scientific framework. This is reflected among other things in the increasing number of paediatric measures and instruments specifically geared to the paediatric rehabilitation profession, for example the Gross Motor Function Measure, Paediatric Evaluation of Disability Inventory and Movement ABC. More recently, paediatric exercise physiologists are pointing to the benefits of an active lifestyle and training for patients with chronic diseases and disabilities. Several studies have evaluated the effects of such training programmes and came up with positive results. It shows that paediatric rehabilitation continues to develop as a dynamic profession, having growth, childhood development and childhood activities as it's core business. PMID- 12745891 TI - Estrogen and progesterone as neuroprotective agents in the treatment of acute brain injuries. AB - Estrogen and progesterone are often thought of as steroid hormones that strongly influence reproductive and maternal behaviours. However, the steroids are now showing considerable promise as neuroprotective and neuroregenerative agents in stroke and traumatic brain injuries. Collectively, these two hormones have been reported to reduce the consequences of the injury cascade by enhancing anti oxidant mechanisms, reducing excitotoxicity: altering glutamate receptor activity, reducing immune inflammation, providing neurotrophic support, stimulating axonal remyelinization and enhancing synaptogenesis and dendritic arborization. Estrogen has often been tried as a prophylactic treatment in females for ischemic brain injury, while progesterone has, thus far, been given as a post-injury treatment for both male and female subjects with acute, ischemic and traumatic injuries of the brain and spinal cord. This review compares and evaluates estrogen and progesterone as neuroactive agents in the acute treatment of brain damage caused by stroke and trauma. PMID- 12745894 TI - The practical use of surface topography: following up patients with Scheuermann's disease. AB - PURPOSE: The purpose of this study was to answer the question of whether a course of in-patient rehabilitation of 4-6 weeks results in a significant postural improvement in patients with a structural kyphosis. Since all clinical measuring parameters are liable to high margins of error and measuring the surface with the formetric system has shown high reliability in previous studies, this system has been used to evaluate the results of rehabilitation in patients with Scheuermann's disease. MATERIALS AND METHOD: A pre-/post-interventional study was undertaken including only patients (n = 136) with Scheuermann's kyphosis and a thoracic curve pattern. 62 female patients with an average age of 19 years and 74 male patients of average age 20 years were diagnosed with the Formetric system before an in-patient rehabilitation programme of 4-6 weeks. For this study, the maximal kyphotic angle was evaluated. RESULTS: The average kyphotic angle as measured by the surface topography system before inpatient rehabilitation was 60.7 degrees (SD 11.2) and after 54.9 degrees (SD 12.1) for the whole sample of 136 patients. The improvements where highly significant in the t-test. In the cases of the female patients, the maximal kyphotic angle decreased from 62 degrees to nearly 54 degrees; in the cases of the male patients it had decreased from 60 degrees to nearly 55 degrees by the end of the programme. DISCUSSION: After the in-patient treatment, the kyphotic angle decreased very significantly. The range of change was far greater than the margin of error and, thus, the following conclusion may be reached: In-patient rehabilitation with an intensive programme of treatment may result in a correction of structural kyphoses in which there are clear signs of Scheuermann's disease. Consequently in-patient rehabilitation is recommended for adolescents in cases of Scheuermann's kyphoses. PMID- 12745893 TI - Resting energy expenditure and body composition in children with myelomeningocele. AB - PURPOSE: Myelomeningocele is a complex disease often complicated by obesity for reasons not well understood. The objectives of this study were to determine body composition and energy expenditure of children with MMC. METHODS: Resting energy expenditure (REE), body composition and anthropometry were measured in 19 children with MMC (12 M, 7 F). Total energy expenditure (TEE) was estimated using a 3-day activity record. Energy intake (EI) was measured in seven children (5 M, 2 F) with MMC. Data were then compared with predicted values. RESULTS: Mean REE (n = 19) was 4680 +/- 1452 kJ/day (96.1 +/- 18.1% of predicted REE). The range was large (45.8-125.7% of predicted REE). TEE (n = 7) was 4344 +/- 2376 kJ/day, hence only 73 34% of predicted TEE. EI (n = 7) was 6560 +/- 1329 kJ/day, approximating a normal energy requirement. Overall, BCM was lower than expected values. CONCLUSIONS: REE in children with MMC is variable when compared to predicted values. TEE was found to be lower in children with MMC than predicted values and EI was similar to predicted values in this group of seven children. BCM is reduced in children with MMC when compared to expected values. PMID- 12745892 TI - Incidence of curvature progression in idiopathic scoliosis patients treated with scoliosis in-patient rehabilitation (SIR): an age- and sex-matched controlled study. AB - The goal of this study is to test the hypothesis that physiotherapy-based intervention can reduce incidence of progression in children with IS. Two independent patient groups matched by age and sex at diagnosis were analysed using the outcome parameter, incidence of progression (> or =5 degrees ). One group was untreated and the other received scoliosis in-patient rehabilitation (SIR). Incidence of progression in groups of untreated patients ranged from 1.5 fold (71.2% vs 46.7%) to 2.9-fold (55.8% vs 19.2%) higher than in groups of patients treated with SIR, even when SIR-treated groups included patients with more severe curvatures. Statistically, the differences were highly significant. Efforts to test the hypothesis that physical therapies addressing postural imbalance can be used effectively in the treatment of IS have been limited. The results of this study are consistent with the possibility that a supervized programme of exercise-based therapies can reduce incidence of progression in children with IS. PMID- 12745895 TI - Cerebral atrophy following shaken impact syndrome and other non-accidental head injury (NAHI). AB - PURPOSE OF THE STUDY: To determine the frequency of cerebral atrophy and microcephaly in a group of children with sequential MRI brain scans after surviving a non-accidental head injury (n = 16). METHODS: Serial head circumference measurements (OFC) were extracted and plotted on standard growth charts for each child retrospectively to determine the frequency of secondary microcephaly. Cerebral atrophy was diagnosed and quantified by measurement of the ventricular/cortical ratio on coronal images of the sequential scans. RESULTS: Acquired microcephaly was found in 15 children (93.8%) over a median follow-up period of 67.93 weeks. There was a significant reduction in the median Z-score for the OFC at the most recent follow-up when compared with that at presentation (p < 0.001, Wilcoxon Signed Rank Test). Cerebral atrophy was found to be the cause of the microcephaly in eight of the 15 children and was evident as early as 9 days after presentation. CONCLUSION: A large proportion of the cohort (93.8%) develops acquired microcephaly after an inflicted head injury and cerebral atrophy is responsible in half of these cases. PMID- 12745896 TI - Commentary on Ylvisaker, M. and Feeney, T.: Executive functions, self-regulation and learned optimism in paediatric rehabilitation: a review and suggestions for intervention. PMID- 12745898 TI - The late endocrine effects of childhood cancer treatment. AB - Whilst many children diagnosed with cancer can now realistically hope for long term survival, the consequences of cancer treatment can be particularly devastating as they enter adolescence and adulthood. Disruption of the endocrine system can result from such treatment, including growth hormone deficiency, problems in normal pubertal progression and thyroid dysfunction. Fertility can also be affected by cancer treatment received as a child, which can have a devastating impact as the patient enters adulthood. In addition, these children may encounter disorders of growth and bone metabolism due to both their initial disease and aspects of its treatment, resulting in further morbidity in later life. The aetiology and diagnosis of these problems are discussed in this review, along with therapeutic options in order to reduce their impact. Long term follow up and clinical vigilance in this patient group is vital. We must continue to strive towards improved survival from childhood cancer, but equally we must remain aware of the adverse effects of treatment, particularly in the long term, and must aim to reduce the impact of these effects as children enter adolescence and adult life. PMID- 12745899 TI - Alexithymia in long-term survivors of childhood cancer. AB - OBJECTIVE: To investigate the incidence of alexithymia (difficulties in describing or recognizing one's own emotions, a limited fantasy life, and general constriction in the affective life) in a group of childhood cancer survivors and to explore medical determinants which predict alexithymia. METHODS: Five years after completing therapy, 72 participants were asked to complete the Bermond Vorst Alexithymia Questionnaire (BVAQ). RESULTS: Male cancer survivors scored significantly lower on overall alexithymia compared to healthy males. They also showed higher ability to fantasize, a higher emotional arousal, and were better able to verbalize their emotional reactions. The female survivors did not show differences compared to the normal female population. No medical determinant was associated with alexithymia. CONCLUSIONS: Stress due to childhood cancer does not affect the alexithymia scores of females. However, male cancer survivors score less alexithymic than age matched controls. PMID- 12745900 TI - Identity and role in parenting a child with cancer. AB - In this paper we suggest that the needs and roles of parents of children with cancer have been inadequately conceptualized by traditional approaches to investigating the psychosocial aspects of chronic childhood illnesses such as cancer. Conducted mainly within discourses of psychopathology, traditional approaches have tended to characterize parents' experience of their child's illness in terms of 'maladjustment' and 'coping', but have done little to illuminate the processes involved in how parents live with their child's illness. In other areas, the research literature treats parents solely as proxy sources of their children's views, and the complexity of their roles as caregivers and individuals in their own right has been ignored. We attempt to re-characterize parenting a child with cancer, drawing attention to how the roles, identities and social obligations of parents position them in relation to the medical world, and highlight the emotional work carried out by parents, including protection of their own and their child's identity. Drawing on various bodies of empirical and theoretical work, including the developing field of childhood studies and the literature on informal carers, we suggest ways of rethinking our understanding of the experience of parenting a child with cancer. In doing so, we consider how the narratives of parents can inform the development of measures to assess the impact of the childhood cancer on the quality of parents' lives, and the role that social and organizational aspects of services can play in ameliorating some of the difficulties involved in parenting a child with cancer. PMID- 12745901 TI - Parenting a child with cancer: promotion and prevention-focused parenting. AB - Applying regulatory focus theory (RFT), it was predicted that, among survivors of childhood cancer, quality of life (QoL) may be compromised by prevention-focused parenting (the focus on avoiding negative outcomes), rather than promotion focused parenting (the focus on approaching positive outcomes). Interviews with mothers of survivors of Acute Lymphoblastic Leukaemia (ALL) and tumours of the Central Nervous System (CNS) were coded for reports of parenting and related to child QoL. Parents reported overall more promotion than prevention; however, mothers of children with tumours of the CNS reported more prevention-focus than mothers of children with ALL. Furthermore, prevention focus was related to child QoL, regardless of diagnosis. The study points toward the value of further development of RFT in clinical contexts. PMID- 12745903 TI - Rehabilitation and development after childhood cancer: can the need for physical exercise be met? AB - Clinical and experimental evidence attests to the neurological and psychological benefits of physical exercise. Prescribing exercise for children after cancer has the potential to facilitate recovery, performance and development, with consequent improvement in quality of life and long-term outcome. However, could a school take up the prescription for a child's individual exercise programme? As a preliminary to an intervention study, all primary and secondary schools in two Scottish regions were surveyed. The aim was to evaluate the utility of prescribing exercise as part of a child's individual rehabilitation plan and to investigate awareness in schools of the potential benefits of exercise for cognition and behaviour. Approximately half the respondents indicated that extra physical exercise sessions could be incorporated into the school day. The main difficulties preventing such inclusion were identified as a lack of staff for supervision and timetabling problems. However, there was also a high level (>70%) of lack of information about what a child could be expected to do after cancer. Further, more respondents (approximately 60%) anticipated benefits of exercise for social development than for academic progress (approximately 35%). These results emphasize the necessity of clear and specific liaison with a school when planning and implementing a child's long-term rehabilitation programme. PMID- 12745902 TI - Reduction of distress associated with paediatric bone marrow transplant: complementary health promotion interventions. AB - Despite numerous medical advances in stem cell/bone marrow transplantation (BMT) for children with malignancies or other catastrophic disorders, BMT remains a prolonged and physically demanding procedure that can be associated with high levels of distress for patients and their families. This paper reviews work from our laboratory that documents the high levels of somatic distress and mood disturbance experienced by patients undergoing the procedure, and points to the need for the development of novel interventions to reduce BMT-related distress. Issues of instrumentation and methodology for outcomes assessment are highlighted. Research is reviewed to support the potential benefits of a number of techniques from the field of complementary medicine (relaxation/imagery; massage, humour therapy, expressive therapy) for reduction of transplant-related distress. Finally, we described a series of studies in which complementary health promotion techniques were assessed in pilot trials. Based on patient and parent perception of helpfulness, two complementary treatments, massage and humour therapy, emerged as the most promising approaches for use in the BMT setting and are now the focus of a randomized clinical trial. PMID- 12745904 TI - Segmentation of renal calculi in ultrasound images. AB - An algorithm proposed by Sridhar and Kumaravel is extended to include a framework for the detection of renal calculi. Calculi occur due to abnormal collection of certain chemicals like oxalate, phosphate and uric acid. These calculi can be present in the kidney, ureter or urinary bladder. Performance analysis is done to a set of five known algorithms using parameters such as success rate in calculi detection, border error metric and time. The framework is constructed by combining the best algorithm based on the performance analysis and a procedure to validate the detected calculi using the shadow it casts in ultrasound images. Ultrasound images of 37 patients are used for testing the algorithm. The detected calculi based on the framework match those determined by expert clinicians in more than 95% of the cases. PMID- 12745905 TI - Psychological cue use and implications for a clinical decision support system. AB - Effective clinical decision making depends upon identifying possible outcomes for a patient, selecting relevant cues, and processing the cues to arrive at accurate judgements of each outcome's probability of occurrence. These activities can be considered as classification tasks. This paper describes a new model of psychological classification that explains how people use cues to determine class or outcome likelihoods. It proposes that clinicians respond to conditional probabilities of outcomes given cues and that these probabilities compete with each other for influence on classification. The model explains why people appear to respond to base rates inappropriately, thereby overestimating the occurrence of rare categories, and a clinical example is provided for predicting suicide risk. The model makes an effective representation for expert clinical judgements and its psychological validity enables it to generate explanations in a form that is comprehensible to clinicians. It is a strong candidate for incorporation within a decision support system for mental-health risk assessment, where it can link with statistical and pattern recognition tools applied to a database of patients. The symbiotic combination of empirical evidence and clinical expertise can provide an important web-based resource for risk assessment, including multi disciplinary education and training. PMID- 12745906 TI - Important variable selection techniques with multiple solutions for medical information applications. AB - The purpose of this study is to utilize the strategy of multiple solutions with easily operated tools to make compatible and affordable trustworthy results. This strategy can satisfy general users in extracting specific knowledge (e.g. diagnosis, treatment, health education, hospital administration, etc.). Wide application is one of the key promoters of a successful medical information system. Risk factors of heart disease can be identified by important variable selection techniques. Four techniques, Chi-square test, correlation analysis, stepwise discriminant analysis, and decision trees, were used in this study. One must compile the results of the different methods to deal with specific research questions in a logical way. It is expected that this approach may reduce the uncertainties obtained from a single method. When applying the important index derived from the four different method results to several applications, all results have been confirmed by current medical knowledge or otherwise provide meaningful information. The prime purpose of this study is not disease diagnosis, but to enable the general health practitioner (medical doctor, hospital administrator, etc.) to formulate their hypothesis. PMID- 12745907 TI - Development of an assessment tool to measure the influence of clinical software on the delivery of high quality consultations. A study comparing two computerized medical record systems in a nurse run heart clinic in a general practice setting. AB - A rating scale was developed to assess the contribution made by computer software towards the delivery of a quality consultation, with the purpose of informing the development of the next generation of systems. Two software programmes were compared, using this scale to test their ability to enable or inhibit the delivery of an ideal consultation with a patient with heart disease. The context was a general practice based, nurse run clinic for the secondary prevention of heart disease. One of the programmes was customized for this purpose; the other was a standard general practice programme. Consultations were video-recorded, and then assessed by an expert panel using the new assessment tool. Both software programmes were oriented towards the implementation of the evidence, rather than facilitating patient-centred practice. The rating scale showed, not surprisingly, significantly greater support from the customized software in the consultation in five out of eight areas. However, the scale's reliability measured by Cronbach's Alpha, was sub-optimal. With further refinement, this rating scale may become a useful tool that will inform software developers of the effectiveness of their programmes in the consultation, and suggest where they need development. PMID- 12745911 TI - Characterization of bones by speckle interferometry. AB - Speckle photography interferometry (SPI) is used to evaluate the stress-strain behaviour of animal bones under loads by recording the speckle fields generated on the bone surface and thus finding the relationship between the fractal dimension D(s) and the compression load P. The procedure consists of application of slow and monotonous compression loads, and recording of the deformation levels and vertical displacements of the specimen. A He-Ne laser beam directed at the surface of bone specimen produces light scattering which generates the speckle. The optical fields show significant differences before and after fracture, due to the changes of the bone surface generated by the strain. Our results support previous suggestions of a relationship between fractal characteristics and parameters related to the mechanical behaviour of materials. PMID- 12745908 TI - A comparison of technologies for database-driven websites for medical education. AB - Static web pages may be easy to setup using text processors or user-friendly web editing software. However, some basic knowledge of the implementation (HTML) is usually needed for final editing and maintenance. As a result many static web pages are left without appropriate updating. With a database driven, dynamic approach web contents may change through user interaction, the pages are usually easier to maintain, and design elements are separated from contents. Database driven solutions (or content management systems) for larger organizations may be purchased at many different levels of complexity. For smaller systems, such as separate courses or projects, they may, however, be developed locally. The present study compares four alternative technologies for database driven web pages. Each technology offers advantages and disadvantages and many issues need to be considered when selecting platform. The present study demonstrates that database driven web pages may be effectively deployed using free Open Source software. Details may be found at www.intermed.dk/datadriven. PMID- 12745912 TI - The utility of neural network in the diagnosis of Cheyne-Stokes respiration. AB - The aim of this study was to design a diagnostic model to identify patients with Cheyne-Stokes respiration (CSR-CSA) based on indices of oximetric spectral analysis. A retrospective analysis of oximetric recordings of 213 sleep studies conducted over a one-year period at a Veterans Affairs medical facility was performed. A probabilistic neural network (PNN) was developed from salient features of the oximetric spectral analysis, desaturation events and the delta index. A fivefold cross-validation was used to assess the accuracy of the neural network in identifying CSR-CSA. When compared to overnight polysomnography, the PNN achieved a sensitivity of 100% (95% confidence interval [CI] 85%-100%) and a specificity of 99% (95% 97%-100%) with a corresponding area under the curve of 99% (95% CI 99%-100%). When combined with overnight pulse oximetry, PNN offers an accurate and easily applicable tool to detect CSR-CSA. PMID- 12745913 TI - Measurement of the symmetry of in vitro stent expansion: a stereo-photogrammetric approach. AB - Balloon-expandable stents are used routinely in the treatment of coronary artery disease. Their effectiveness is limited by the occurrence of restenosis. Previous studies have suggested that the level of restenosis may be related to the deployed stent geometry, and in particular to the symmetry of the deployment profile. It is suggested that the symmetry of deployment might be influenced by the folding pattern of the balloon on which the stent is delivered. This paper describes a stereo-photogrammetric system for the three-dimensional reconstruction of stent geometry during expansion, including appropriate specification and calibration procedures. Calibration testing of the system indicated an accuracy of +/-0.05 mm in the reconstruction of the position of a point on the stent surface. Methods for processing the 3D data are described, including a technique for quantitatively differentiating between results from two alternative balloon folding patterns. This study may aid future balloon and stent design with respect to the optimization of stent deployment characteristics. PMID- 12745914 TI - From wired to wireless: a miniature robot for intestinal inspection. AB - Increasingly more robots are used for various medical purposes, one of which is for intestinal inspection. A prototype robotic endoscope is presented along with a locomotion mechanism and relative experiment. Image, pH, pressure and temperature, etc. can be measured by the robot, which is very helpful in diagnoses. A wired robot will be replaced by a wireless robot because it is not comfortable for subjects and the wired robot cannot advance further into the small intestine because of too many folds and projections in the intestine. Key obstacles of the future wireless robot for intestinal inspection are discussed in detail and a reversed tracking method is provided. PMID- 12745915 TI - Respiratory effect on the pulse spectrum. AB - During the last decade, progressive achievements in haemodynamics have shown that radial artery pulse pressure can be used to estimate the vascular properties of the internal organs. Clinical experiments have shown that slow and regular respiration has a large effect on the heart rate variability (HRV). This phenomenon is called respiratory sinus arrhythmia (RSA). It is known that respiration-related oscillations in venous return cause oscillations in stroke volume and blood pressure. It also can be inferred from cardiac output that systemic blood pressure has a similar respiration-related cycle. Moreover we found that the fluctuations of harmonics of arterial pulse are consistent with the fluctuation of HRV. This means that the whole cardiovascular system (CVS) makes rapid adaptation during respiration, and the harmonic proportions of arterial pulse were modified during different breath rates. This result shows that the regular respiration also has a large effect upon Windkessel properties of CVS. PMID- 12745916 TI - Simulation of systolic and diastolic left ventricular dysfunction in a mock circulation: the effect of arterial compliance. AB - Arterial compliance (AC) is expected to play a major role on cardiac efficacy by acute or long-term mechanisms. The aim of this study was to investigate the purely mechanical effect of AC on left ventricular (LV) performance, for different conditions of LV dysfunction (systolic versus diastolic). A hydraulic, Windkessel model of systemic circulation was used. LV function and aortic flow were simulated using a left ventricular assist device (LVAD). Two cases of LV dysfunction were simulated: Case A, systolic and Case B, diastolic dysfunction. In Case A, AC increased from 1.14 to 2.85 ml mm Hg(-1) leading to an increase in LVAD stroke volume up to 6%, while no significant effect was observed in Case B. LVAD systolic work was decreased by 4% in systolic and by 11% in diastolic LVAD dysfunction. The purely mechanical effect of AC changes on LVAD function was different between systolic and diastolic dysfunction. It might be expected that even an acute reduction in arterial stiffness could enhance LV performance by different means in systolic compared to diastolic dysfunction. PMID- 12745919 TI - The fence and picket structure of the plasma membrane of live cells as revealed by single molecule techniques (Review). AB - Models of the organization of the plasma membrane of live cells as discovered through diffusion measurements of integral membrane molecules (transmembrane and GPI-anchored proteins, and lipid) at the single molecule level are discussed. Diffusion of transmembrane protein and, indeed, even lipid is anomalous in that the molecules tend to diffuse freely in limited size compartments, with infrequent intercompartment transitions. This average residency time in a compartment is dependent on the diffusing species and on its state of oligomerization, becoming completely confined to a single compartment upon sufficient oligomerization. This will be of great importance in determining cellular mechanisms for controlling the random diffusive motion of membrane molecules and in understanding signalling processes. PMID- 12745920 TI - Peptidase allergen Der p 1 initiates apoptosis of epithelial cells independently of tight junction proteolysis. AB - Loss of epithelial cell polarity, which can arise following disruption of tight junctions (TJs), is a precursor to the care-fully orchestrated removal of moribund cells from epithelia in apoptosis. Ordinarily, this cycle of events has minimally disruptive effects on the function of the epithelial barrier, but some agents have been identified that induce apoptosis and promote epithelial leakiness. The allergen Der p 1 is a cysteine peptidase that cleaves TJ adhesion proteins and induces apoptosis in epithelial cells. This suggests the possibility that, at least for some inducers of apoptosis, these events might be causally linked. We report here that Der p 1 induces epithelial apoptosis before outright cell detachment and that apoptosis occurs within the same time span as increased paracellular permeability in polarized epithelial monolayers. Whilst TJ-deficient BEAS-2B cells were resistant to Der p 1-induced apoptosis, the cell line 1HAEo-, which was also TJ deficient, was sensitive to Der p 1, providing evidence against TJ proteolysis as a cause of apoptosis. To provide direct evidence, we propagated cells that normally express TJs in low calcium medium that prevented intercellular junction assembly. These cells retained full susceptibility to Der p 1, indicating that Der p 1-induced apoptosis is independent from TJ proteolysis. PMID- 12745921 TI - Challenging accepted ion channel biology: p64 and the CLIC family of putative intracellular anion channel proteins (Review). AB - Parchorin, p64 and the related chloride intracellular channel (CLIC) proteins are widely expressed in multicellular organisms and have emerged as candidates for novel, auto-inserting, self-assembling intracellular anion channels involved in a wide variety of fundamental cellular events including regulated secretion, cell division and apoptosis. Although the mammalian phosphoproteins p64 and parchorin (49 and 65K, respectively) have only been indirectly implicated in anion channel activity, two CLIC proteins (CLIC1 and CLIC4, 27 and 29K, respectively) appear to be essential molecular components of anion channels, and CLIC1 can form anion channels in planar lipid bilayers in the absence of other cellular proteins. However, these putative ion channel proteins are controversial because they exist in both soluble and membrane forms, with at least one transmembrane domain. Even more surprisingly, soluble CLICs share the same glutaredoxin fold as soluble omega class glutathione-S-transferases. Working out how these ubiquitous, soluble proteins unfold, insert into membranes and then refold to form integral membrane proteins, and how cells control this potentially dangerous process and make use of the associated ion channels, are challenging prospects. Critical to this future work is the need for better characterization of membrane topology, careful functional analysis of reconstituted and native channels, including their conductances and selectivities, and detailed structure/function studies including targeted mutagenesis to investigate the structure of the putative pore, the role of protein phosphorylation and the role of conserved cysteine residues. PMID- 12745922 TI - Structure and function of potassium channels in plants: some inferences about the molecular origin of inward rectification in KAT1 channels (Review). AB - Potassium channels in plants play a variety of important physiological roles including K(+) uptake into roots, stomatal and leaf movements, and release of K(+) into the xylem. This review summarizes current knowledge about a class of plant genes whose products are K(+) channel-forming proteins. Potassium channels of this class belong to a superfamily characterized by six membrane-spanning domains (S1-6), a positively charged S4 domain and a region between the S5 and S6 segments that forms the channel selectivity filter. These channels are voltage dependent, which means the membrane potential modifies the probability of opening (P(o)). However, despite these channels sharing the same topology as the outward rectifying K(+) channels, which are activated by membrane depolarization, some plant K(+) channels such as KAT1/2 and KST1 open with hyperpolarizing voltages. In outward-rectifying K(+) channels, the change in P(o) is achieved through a voltage sensor formed by the S4 segment that detects the voltage transferring its energy to the gate that controls pore opening. This coupling is achieved by an outward displacement of the charges contained in S4. In KAT1, most of the results indicate that S4 is the voltage sensor. However, how the movement of S4 leads to opening remains unanswered. On the basis of recent data, we propose here that in plant-inward rectifiers an inward movement of S4 leads to channel opening and that the difference between it and outward-rectifying channels resides in the mechanism that couples gating charge displacement with pore opening. PMID- 12745923 TI - Understanding ATP synthesis: structure and mechanism of the F1-ATPase (Review). AB - To couple the energy present in the electrochemical proton gradient, established across the mitochondrial membrane by the respiratory chain, to the formation of ATP from ADP and Pi, ATP-synthase goes through a sequence of coordinated conformational changes of its major subunits (alpha, beta). These changes are induced by the rotation of the gamma subunit driven by the translocation of protons through the c subunit of the membrane portion of the enzyme. During this process, the F1-portion of the ATP-synthase adopts at least two major conformations depending on the occupancy of the beta subunits: one with two nucleotides, the other with three. In the two-nucleotide structure, the empty beta subunit adopts an open conformation that is highly different from the other conformations of beta subunits: tight, loose and closed. The three-dimensional structures of the F1-ATPase in each of these two major conformations provide a framework for understanding the mechanism of energy coupling by the enzyme. The energetics associated with two different models of the reaction steps, analysed using molecular dynamics calculations, show that three-nucleotide intermediates do not occur in configurations with an open beta subunit; instead, they are stabilized by completing a jaw-like motion that closes the beta subunit around the nucleotide. Consequently, the energy driven, major conformational change takes place with the beta subunits in the tight, loose and closed conformation. PMID- 12745924 TI - Lipoplex size determines lipofection efficiency with or without serum. AB - In order to identify factors affecting cationic liposome-mediated gene transfer, the relationships were examined among cationic liposome/DNA complex (lipoplex) cell interactions, lipoplex size and lipoplex-mediated transfection (lipofection) efficiency. It was found that lipofection efficiency was determined mainly by lipoplex size, but not by the extent of lipoplex-cell interactions including binding, uptake or fusion. In addition, it was found that serum affected mainly lipoplex size, but not lipoplex-cell interactions, which effect was the major reason behind the inhibitory effect of serum on lipofection efficiency. It was concluded that, in the presence or absence of serum, lipoplex size is a major factor determining lipofection efficiency. Moreover, in the presence or absence of serum, lipoplex size was found to affect lipofection efficiency by controlling the size of the intracellular vesicles containing lipoplexes after internalization, but not by affecting lipoplex-cell interactions. In addition, large lipoplex particles showed, in general, higher lipofection efficiency than small particles. These results imply that, by controlling lipoplex size, an efficient lipid delivery system may be achieved for in vitro and in vivo gene therapy. PMID- 12745925 TI - Extent of the selectivity filter conferred by the sixth transmembrane region in the CFTR chloride channel pore. AB - Point mutations within the pore region of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel have previously been shown to alter the selectivity of the channel between different anions, suggesting that part of the pore may form an anion 'selectivity filter'. However, the full extent of this selectivity filter region and the location of anion binding sites in the pore are currently unclear. As a result, comparisons between CFTR and other classes of Cl( ) channel of known structure are difficult. We compare here the effects of point mutations at each of eight consecutive amino acid residues (arginine 334-serine 341) in the crucial sixth transmembrane region (TM6) of CFTR. Anion selectivity was determined using patch-clamp recording from inside-out membrane patches excised from transiently transfected mammalian cell lines. The results suggest that selectivity is predominantly controlled by a single site involving adjacent residues phenylalanine 337 and threonine 338, and that the selectivity conferred by this 'filter' region is modified by anion binding to flanking sites involving the more extracellular arginine 334 and the more intracellular serine 341. Other residues within this part of the pore play only minor roles in controlling anion permeability and conductance. Our results support a model in which specific TM6 residues make important contributions to a single, localized anion selectivity filter in the CFTR pore, and also contribute to multiple anion binding sites both within and on either side of the filter region. PMID- 12745926 TI - Neither lipophilicity nor membrane-perturbing potency of phenothiazine maleates correlate with the ability to inhibit P-glycoprotein transport activity. AB - Although phenothiazines are known as multidrug resistance modifiers, the molecular mechanism of their activity remains unclear. Since phenothiazine molecules are amphiphilic, the interactions with membrane lipids may be related, at least partially, to their biological effects. Using the set of phenothiazine maleates differing in the type of phenothiazine ring substitution at position 2 and/or in the length of the alkyl bridge-connecting ring system and side chain group, we investigated if their ability to modulate the multidrug resistance of cancer cells correlated with model membrane perturbing potency. The influence exerted on lipid bilayers was determined by liposome/buffer partition coefficient measurements (using the absorption spectra second-derivative method), fluorescence spectroscopy and calorimetry. Biological effects were assessed by a flow cytometric functional test based on differential accumulation of fluorescent probe DiOC(2)(3) by parental and drug-resistant cells. We found that all phenothiazine maleates were incorporated into lipid bilayers and altered their biophysical properties. With only few exceptions, the extent of membrane perturbation induced by phenothiazine maleates correlated with their lipophilicity. Within the group of studied derivatives, the compounds substituted with CF(3)- at position 2 of phenothiazine ring were the most active membrane perturbants. No clear relation was found between effects exerted by phenothiazine maleates on model membranes and their ability to modulate P-glycoprotein transport activity. PMID- 12745927 TI - PTS1-independent targeting of isocitrate lyase to peroxisomes requires the PTS1 receptor Pex5p. AB - The targeting of castor bean isocitrate lyase to peroxisomes was studied by expression in the heterologous host Saccharomyces cerevisae from which the endogenous ICL1 gene had been removed by gene disruption. Peroxisomal import of ICL was dependent upon the PTS1 receptor Pex5p and was lost by deletion of the last three amino acids, Ala-Arg-Met. However, removal of an additional 16 amino acids restored the ability of this truncated ICL to be targeted to peroxisomes and this import activity, like that of the full-length protein, was dependent upon Pex5p. The ability of peptides corresponding to the carboxyl terminal ends of wild-type and Delta 3 and Delta 19 mutants of ICL to interact with the PTS1 binding portion of Pex5p from humans, plants and yeast was determined using the yeast two-hybrid system. The peptide corresponding to wild-type ICL interacted with all three Pex5p proteins to differing extents, but neither mutant could interact with Pex5p from any species. Thus, ICL can be targeted to peroxisomes in a Pex5p-dependent but PTS1-independent fashion. These results help to clarify the contradictory published data about the requirement of the PTS1 signal for ICL targeting. PMID- 12745928 TI - Influence of erythrocyte shape on the rate of Ca2+-induced scrambling of phosphatidylserine. AB - Membrane-perturbing agents that cause transformation of biconcave erythrocytes into echinocytes or stomatocytes were used to investigate the influence of erythrocyte shape on the rate of Ca(2+)-induced scrambling of phospholipids. Erythrocytes were treated with a variety of lipid-soluble compounds to induce these shape changes, followed by incubation with calcium and ionomycin to activate lipid scramblase. Prothrombinase activity of the cells was used to monitor the rate of surface exposure of phosphatidylserine, which is taken as a measure of scramblase activity. Echinocytes show an enhanced rate of scrambling, whereas stomatocytes show a reduced rate, relative to normocytes. This phenomenon appears to correlate with enhanced and diminished micro-exovesicle shedding from echinocytes and stomatocytes, respectively. It is concluded that the rate of calcium-induced phosphatidylserine exposure (rate of lipid scrambling) in erythrocytes depends for a considerable part on the cells' ability to form microvesicles. PMID- 12745932 TI - Delayed auditory feedback in the treatment of stuttering: clients as consumers. AB - PURPOSE: To investigate the effect of repeated exposure to delayed auditory feedback (DAF) during a 3-month period outside a clinical environment and with only minimal clinical guidance on speech fluency in people who stutter. METHOD: A pretest-post-test design was used with repeated exposure to DAF during 3 months as the independent variable. Nine stuttering individuals aged between 18 and 45 years served as subjects. Videotaped samples from different speech tasks were collected before and after 3 months' exposure to DAF, each time under two conditions: first during non-altered feedback (NAF) and subsequently during DAF. RESULTS: Before the repeated exposure to DAF, the percentage of stuttered words was significantly higher during NAF than during DAF. After 3 months' exposure to DAF, the percentage of stuttered words during NAF had dropped to a non significant level in comparison with the DAF condition. Comparison of the percentage of stuttered words during NAF before and after repeated exposure to DAF showed significantly less stuttering after the repeated exposure to DAF. The percentage of stuttered words during DAF was generally somewhat higher, but not significantly so, after the 3 months' repeated exposure to DAF than before the exposure to DAF. CONCLUSIONS: Results confirm that DAF is an effective means of reducing stuttering even when employed as the only treatment approach outside a therapeutic environment. They also suggest that DAF continues to promote fluency when used over a longer period, but also that length of exposure and fluency enhancement are probably not in a linear relationship. PMID- 12745933 TI - Writing with voice: an investigation of the use of a voice recognition system as a writing aid for a man with aphasia. AB - BACKGROUND: People with aphasia may experience difficulties that prevent them from demonstrating in writing what they know and can produce orally. Voice recognition systems that allow the user to speak into a microphone and see their words appear on a computer screen have the potential to assist written communication. AIM: This study investigated whether a man with fluent aphasia could learn to use Dragon NaturallySpeaking to write. METHODS & PROCEDURES: A single case study of a man with acquired writing difficulties is reported. A detailed account is provided of the stages involved in teaching him to use the software. The therapy tasks carried out to develop his functional use of the system are then described. Outcomes included the percentage of words accurately recognized by the system over time, the quantitative and qualitative changes in written texts produced with and without the use of the speech-recognition system, and the functional benefits the man described. OUTCOMES & RESULTS: The treatment programme was successful and resulted in a marked improvement in the subject's written work. It also had effects in the functional life domain as the subject could use writing for communication purposes. CONCLUSIONS: The results suggest that the technology might benefit others with acquired writing difficulties. PMID- 12745934 TI - Training volunteers as conversation partners for people with aphasia. AB - BACKGROUND: One of the most disabling consequences of aphasia is the way it excludes the person from conversation. A number of studies have attempted to tackle this problem by training the conversational partners of aphasic people. This study offers an evaluation of this approach. AIMS: Six volunteers were trained in conversing with people with moderate or severe aphasia. The study aimed to evaluate whether training changed the volunteers' knowledge about aphasia and their interactions with aphasic people. It also explored whether changes in the volunteers were matched by improved participation of the aphasic people in conversation. METHODS & PROCEDURES: Volunteers were recruited from an aphasia group in Milton Keynes. They were trained as a group over three sessions (9 hours in total). The training drew on the techniques of Kagan (1998, 1999). It included presentation of information using different media, group discussions, viewing of videos and role play. The course aimed to improve (1) the volunteers' understanding of the nature of aphasia, (2) the volunteers' knowledge about communication strategies to use with aphasic people and (3) the skills of the volunteers in supporting aphasic people in conversation. An additional aim was to increase the aphasic subjects' participation in conversation. Two evaluation methods were employed. Specially designed questionnaires were administered before and after training to evaluate the volunteers' knowledge and understanding. Volunteers were also videotaped in conversation with aphasic people before and after training. The videos were rated by speech and language therapists, using nine-point rating scales, derived from Kagan (1999). OUTCOMES & RESULTS: Significant improvements were seen on the questionnaire scores and in ratings of the volunteers' videos. The rating gains were attributed to the training course, since baseline scores were stable and improvements only occurred after the training period. There were comparable gains in the participation of the aphasic subjects, which again occurred after training. CONCLUSIONS: The study demonstrates that a short training course can change the knowledge and practice of experienced volunteers. The findings have implications for teaching generic skills to volunteers working with aphasic people. PMID- 12745935 TI - Predicting treatment time with the Lidcombe Program: replication and meta analysis. AB - BACKGROUND: The benefits of treating stuttering close to onset have become obvious in recent years, and the Lidcombe Program has emerged as an effective and safe treatment method for children in their preschool years. The benefits of implementing the programme with young children, however, need to be weighed against the knowledge that many children recover from stuttering without treatment. In light of this, speech-language therapists need to know how long treatment is likely to take and whether treatment time can be predicted. In particular, they need to know if adopting a 'watchful waiting' approach--to see if natural recovery occurs--jeopardizes responsiveness to treatment. A recent Australian study of 250 preschool-age children found that stuttering rate was the only significant predictor of treatment time with the Lidcombe Program. In other words, children whose stuttering was more severe took longer to pass through the programme. There were other trends in the data but they did not reach significance. AIMS: The present study, conducted independently in the UK, was designed to replicate the Australian study. Direct replication enabled pooling of the data from the two studies in a meta-analysis. METHODS & PROCEDURES: The study included 66 children who began treatment before 6 years of age. They were treated with the Lidcombe Program at a specialist stuttering clinic in Norwich. Logistic regression analyses were conducted on the data. The data from both the British and Australian cohorts were pooled in a meta-analysis. OUTCOMES & RESULTS: Results indicated that Stage 1 of the Lidcombe Program was completed in a median of 11 clinic visits, which is in line with the findings of the Australian study. Stuttering rate at first clinic visit was again found to be a significant predictor of treatment time. The remaining data trends were similar to those in the Australian data. In the meta-analysis, stuttering rate was once more found to be a predictor of treatment time. Of particular interest, however, was that the increased power provided by the meta-analysis identified an additional predictor, namely onset-to-treatment interval. Contrary to what is known about the responsiveness of children to the Starkweather and Gottwald treatment, and contrary to what might be expected given what we know about natural recovery, children who had been stuttering for more than 12 months took less time to progress through the programme than children who had been stuttering for less than 12 months. CONCLUSIONS: These findings indicate that delaying intervention with the Lidcombe Program for 1 year after onset, within the preschool years, is unlikely to jeopardize responsiveness to treatment. The clinical implications of these findings are discussed. PMID- 12745936 TI - Relationship between gestures and words in children with Down's syndrome and typically developing children in the early stages of communicative development. AB - BACKGROUND: Previous research has emphasized the importance of gesture in early communicative development. These studies have reported that gestures are used frequently during the first two years of life and may play a transitional role in the language acquisition process. Although there are now numerous descriptions of the relationship between gesture and the developing language system in typically developing (TD) children, relatively little is known about the nature and early development of the gesture-language system in children with developmental disorders involving specific profiles of language delay and/or impairment. PRIMARY OBJECTIVE: The aim of this study is to compare early word and gesture use in children with DS and in typically-developing children to investigate potential differences in the relationship between gestural and verbal communication in early language development. METHODS AND PROCEDURES: Ten children from upper middle class families participated in the study. The five children with DS (3 boys and 2 girls) had an average chronological age of 47.6 months, an average mental age of 22.4 months, and an average language age of 18 months. Each child with DS was matched to a typically developing child on the basis of gender, language age, and observed expressive vocabulary size. Children were videotaped for 30 minutes as they interacted spontaneously with their mothers. All communicative and intelligible gestures and words produced by the children were transcribed from the videotapes. Data analyses focused on: a) overall production of gestures and words (i.e., gesture and word tokens); b) the size of children's gestural and verbal repertoires (i.e., gesture and word types); and c) production and informational content of gesture-word combinations. MAIN OUTCOMES AND RESULTS: Although children with DS had significantly smaller gestural repertoires than their language age-matched peers, there was no reliable difference between the two groups in the overall use of gesture. In addition, with DS produced two element combinations (primarily gesture-word combinations) and did so at a rate comparable to that observed among their TD counterparts. However, no two-word combinations were observed among children with DS, and there were also group differences in the information contained in children's gesture-word combinations. CONCLUSIONS: Taken together, these findings suggest that in addition to the well documented global delays in early communicative development, children with DS may exhibit additional pockets of delay, specifically in making the transition from one- to two-word speech. Results are further discussed in terms of their implications for understanding the organization of the developing gesture language system and for the assessment of gesture in young children with communicative delays and disorders. PMID- 12745938 TI - Reliability of scores between stroke patients and significant others on the Reintegration to Normal Living (RNL) Index. AB - PURPOSE: This study measured reliability between stroke patients' and significant others' scores on items on the Reintegration to Normal Living (RNL) Index and whether there were any scoring biases. METHOD: The 11-item RNL Index was administered to 57 pairs of patients and significants six months after stroke rehabilitation. The index was scored using a 10-point visual analogue scale. Patient and significant other demographic information and data on patients' clinical, functional and cognitive status were collected. Reliability was measured using the intra-class correlation coefficient (ICC) and percent agreement. RESULTS: Overall poor reliability was found for the RNL Index total score (ICC=.36, 95% CI .07 to .59) and the daily functioning subscale (ICC=.24, 95% Cl -.003 to .46) and moderate reliability was found for the perception of self subscale (ICC= .55, 95% Cl .28 to .73). There was a moderate bias for patients to rate themselves as achieving better reintegration than was indicated by significant others, although no demographic or clinical factors were associated with this bias. Exact match agreement was best for the subjective items and worse for items reflecting mobility around the community and participation in a work activity. CONCLUSIONS: Caution is needed when interpreting patient information reported by significant others on the RNL Index. The use of a shorter scale to rate the RNL Index requires investigation. PMID- 12745939 TI - The coping responses of the adolescent siblings of children with severe disabilities. AB - PURPOSE: There is a paucity of literature conducted on adolescents' perceptions of their siblings with severe disabilities. The period of adolescence is characterized by personal and emotional conflicts resulting from teenagers' search towards self-identity and autonomy and exploration of the boundaries of parental support. The study attempts to describe the coping responses of these adolescent siblings in their adjustment to the family stressor of having a sibling with a disability; and the adolescents' available coping resources. METHOD: This study is qualitative in nature and open-ended, structured interviews were conducted with 19 adolescents between the ages of 12 and 15 years who had a sibling with severe disabilities. The interviews were then analysed according to categories using an editing analysis style. RESULTS: The results indicated that the subjects reported limited family interaction and were often reticent in expressing their feelings about their sibling with a disability. In addition, they expressed guilt feelings regarding their siblings with disabilities. Furthermore, siblings of children with severe disabilities received limited information and guidance regarding their sibling's disability. CONCLUSION: The need for professional support to facilitate adolescents' coping with the disability and its consequences as well as the establishment of strong support networks for these adolescents became evident. PMID- 12745940 TI - Physiotherapy practice in stroke rehabilitation: a survey. AB - OBJECTIVE: This survey aimed to provide an expert consensus view of the theoretical beliefs underlying physiotherapy practice in stroke rehabilitation the United Kingdom. METHOD: Questionnaires (with sections related to: therapist background, physiotherapy management, theoretical beliefs and gait re-education strategies used) were posted to all senior level physiotherapists working in stroke care (n=1022). RESULTS: The majority of respondents had more than 10 years experience overall, and at least 5 years experience in stroke care. The Bobath concept was the preferred approach (n=67%) followed by an 'eclectic' approach (n=31%). This survey identified four theoretical themes underlying current practice in neurological physiotherapy: the promotion of normal movement, the control of tone, the promotion of function, and the recovery of movement with optimization of compensation. CONCLUSIONS: A consensus was obtained on 16 theoretical beliefs; however the evidence base underlying these beliefs remains sparse. Many of these beliefs require further debate within the physiotherapy profession such as the amount of time spent on preparation for function, the automatic translation of movement into function, carry over outside therapy, and the way in which tasks should be practised. PMID- 12745941 TI - Development and reliability of the General Motor Function Assessment Scale (GMF)- a performance-based measure of function-related dependence, pain and insecurity. AB - PURPOSE: To develop a scale for assessment of three components-dependence, pain and insecurity - related to motor functions of importance for activities of daily living among older rehabilitation patients and to establish its clinical practicality and reliability. METHOD: A General Motor Function Assessment Scale (GMF) with the above aims was constructed. Clinical practicality was explored by questionnaires to 14 physiotherapists. Inter-rater and test-retest reliability was tested on patients in three different forms of geriatric rehabilitation (n=20 25) and analysed by percentage agreement (PA) and a non-parametric statistical method, which provide measures of the random disagreement separately from the systematic part of the disagreement. RESULTS: In the clinical test the GMF was found to be time efficient and clinically adequate. Analysis of reliability showed overall high values of PA (PA> or =70) and of the rank-order agreement coefficient (r(a)>0.82), and low degrees of systematic disagreement. CONCLUSIONS: GMF was found to be a clinically useful assessment scale in geriatric rehabilitation. The statistical analyses indicted a high degree of reliability. Comparison of these results with reliability of comparable rating scales is difficult on account of the statistical methods used in other studies, which commonly do not take into account the non-metric properties of the data. PMID- 12745942 TI - Assessing unilateral neglect: shortcomings of standard test methods. AB - INTRODUCTION: When investigating the incidence of unilateral neglect in a first ever stroke population, we found that some patients showed clinical signs of neglect, but managed to pass our tests. The purpose of this paper is to describe the nature of such signs, and analyse why test instruments were insufficiently corresponding to those signs. METHOD: One hundred and thirty-one consecutive patients with first-ever stroke in a community-based sample were evaluated for the presence of unilateral neglect. We used a test battery consisting of tests for visuo-spatial neglect, personal neglect, and anosognosia. Twenty cases of neglect were discovered by standard methods. We asked our collaborators at the wards to report any behavioural abnormality reminiscent of neglect present in patients who had normal test results. Such patients were evaluated clinically. RESULTS: Nine cases with neglect-like symptoms were discovered. Our clinical evaluation of the nine patients indicated several possible explanations for their behavioural abnormalities, including motor neglect, neglect for far extrapersonal space, disturbances of proprioception, and spatial disturbances other than neglect. CONCLUSION: Standard neglect tests do not cover all clinical forms of neglect. It is therefore important not to rely completely on test instruments when diagnosing neglect. More versatile test instruments are desired. PMID- 12745943 TI - Health related quality of life and related factors in 539 persons with amputation of upper and lower limb. AB - PURPOSE: Limb amputation is followed by an important rehabilitation process, especially when a prosthesis is involved. The objective of this study is to assess the nature of factors related to health related quality of life (HRQL) of persons with limb amputation. METHOD: The Nottingham Health Profile (NHP) treated 1011 subjects with major amputation of one or several limbs. Correlations were sought in multivariate regression model analyses between the six categories of distress explored by the NHP and age, sex, cause and level of amputation and rehabilitation programme. RESULTS: Response rate was 53.3%. HRQL measured by the NHP was mostly impaired in the categories of physical disability, pain and energy level. Controlling for sex and age, young age at the time of amputation, traumatic origin and upper limb amputation were independently associated with better HRQL. CONCLUSION: It is concluded that HRQL is largely related to factors which are inherent to the patient and the amputation. PMID- 12745944 TI - Rates of access to assistive equipment and medical rehabilitation services among people with disabilities. AB - PURPOSE: To determine rates of access to assistive equipment and medical rehabilitation services among people with disabilities in the US, and to determine whether health plan type is associated with rates of access to these health services. METHOD: Results were derived from a nationwide US survey sample of people with cerebral palsy, multiple sclerosis or spinal cord injury. Analyses were restricted to working-age adults (n=500). Need for, and receipt of (1) assistive equipment in the last 12 months, and (2) rehabilitative services in the last 3 months, was determined. RESULTS: Over half of the sample indicated a need for assistive equipment in the last 12 months. Nearly a third of those who indicated a need did not receive assistive equipment every time it was needed. Forty per cent of the sample indicated a need for rehabilitative services in the last 3 months, and over half of those indicating a need did not receive rehabilitative services every time they were needed. Access rates did not differ appreciably between respondents covered by managed care and fee-for-service health plans. CONCLUSION: Emphasis in healthcare for people with disabilities should shift from traditional acute healthcare models that focus on functional restoration, to preventive services, and maintenance of function, health and independence. PMID- 12745945 TI - Self-perceptions of speech language pathologists-in-training before and after pseudostuttering experiences on the telephone. AB - PURPOSE: This survey investigated the effect of 'pseudostuttering' experiences on self-perceptions of 29 female, graduate students enrolled in a graduate seminar in stuttering while in a programme of study to become professional speech language pathologists. METHOD: Perceptions of self prior to, and immediately after, participation in five scripted telephone calls that contained pseudostuttering were measured via a 25-item semantic differential scale. RESULTS: Participants perceived themselves as significantly more (p < 0.002) withdrawn, tense, avoiding, afraid, introverted, nervous, self-conscious, anxious, quiet, inflexible, fearful, shy, careless, hesitant, uncooperative, dull, passive, unpleasant, insecure, unfriendly, guarded, and reticent after their pseudostuttering telephone call experiences. CONCLUSIONS: Findings suggests that the pseudostuttering experiences have an impact on self-perceptions and that the experience of 'adopting the disability of a person who stutters' may provide insight as to the social and emotional impact of communicative failure. It is suggested that pseudostuttering exercises may be a valuable teaching tool for the graduate students, especially for those who do not stutter. PMID- 12745946 TI - Getting the work done: a qualitative study of individuals with multiple sclerosis. AB - THE PROBLEM: Work can be defined as an activity performed to accomplish something in the presence of obstacles that may make accomplishing the goal difficult. For individuals with MS, work is not only limited by physical impairments but also by factors such as fatigue and cognitive changes PURPOSE: The aim of this study is to examine the experiences of individuals with mild to moderate MS as they carry out everyday work activities both inside and outside the home. METHOD: Eleven women and three men were recruited from the community to participate in a series of semi-structured interviews. Using qualitative research methodology that examined the experiences of the participants, two major themes and seven subthemes emerged. RESULTS: The first theme, Defining the work, included Priorities: seeing what's important; Plans: learning about resources and requirements; and PERSPECTIVES: fixing it yourself. The second theme, Changing how things get done, includes Precipitating factors; Awareness; Constructing the strategies; and Evaluating the strategies. CONCLUSIONS: Individuals with MS develop strategies and utilize resources in order to get the work done. Comparisons are made between existing intervention theories or programmes and the experiences described by participants in this study. PMID- 12745947 TI - Disability advocacy organizations: a descriptive framework. AB - PURPOSE: The purpose of this paper is to offer a descriptive framework to analyse the participation of people with disabilities in policy advocacy over the past 20 years. METHODS: The study used a qualitative research design to describe and analyse organizations participating in policy advocacy on behalf of people with disabilities in Canada between 1981 and the present. Data for the study were assembled through interviews with key informants who had participated in three policy cases: the Charter of Rights and Freedoms, 1982 (federal); the Canada Clause of the Charlottetown Accord, 1991 (federal); the Ontario Consent to Treatment, Advocacy and Substitute Decision-making bills, 1993-1995 (provincial). For each of the three cases, key informants were identified who could report with authority on the participation of individuals or groups representing people with disabilities. Interviews with key informants were conducted by the investigators, transcribed and coded to produce five dimensions along which advocates described their organizations. RESULTS: Three ideological dimensions were identified to describe organizations representing people with disabilities: purpose, tactics, and view of disability. Two practical dimensions were also identified: membership and resources. Each of these dimensions is described in detail with quotes from the interviews to illustrate. DISCUSSION AND CONCLUSIONS: These five dimensions result in a descriptive framework for discussion and comparison of organizational characteristics of advocacy organizations representing people with disabilities. Further, the framework affords a vehicle for discussion of the relative success of policy participation on issues affecting the lives of people with disabilities. PMID- 12745948 TI - The possession of technical aids among persons with a somatic chronic disease. AB - PURPOSE: Previous research has highlighted disability as a determinant of the need for technical aids; surprisingly, disease as a potential determinant has been ignored. The goal of the present study was to determine whether the possession of technical aids is dependent on the type of chronic disease, illness duration, co-morbidity, disability, age, and several other factors. METHODS: The study was performed in a representative sample of persons with a somatic chronic disease (n=2262). Type of chronic disease, time post-diagnosis and co-morbidity were assessed by the patient's general practitioner. Disability was assessed with the Sickness Impact Profile. The possession of technical aids and other characteristics were assessed by questionnaire. RESULTS: Type of chronic disease (osteoarthritis, rheumatoid arthritis, diabetes and COPD), disability, age and gender were significantly and independently associated with the possession of technical aids. CONCLUSIONS: Apart from disability, disease-, age-, and gender related characteristics determine the possession of technical aids. A detailed assessment of these characteristics is required in order to ensure that technical aids meet personal needs. PMID- 12745950 TI - Process and outcome during early inpatient rehabilitation after brain injury. AB - PURPOSE: To describe aspects of process and outcome during early inpatient rehabilitation of younger adults after single incident brain injury. METHOD: Analysis of a database of 290 patients discharged from an inner-city hospital based inpatient unit for younger adults after single incident neurological events, over a 5-year period. RESULTS: Analysis showed a stable case-mix of patients over the 5 years surveyed with a preponderance of young male patients after traumatic brain injury. Improvements in patients' disability and dependency, measured by the Barthel Index and Functional Independence Measure, were recorded in the majority of patients. The ethnic diversity of the patients, reflecting the multi-cultural nature of the catchment population, did not appear to affect rehabilitation outcome. Failure to record improvement in 15% of patients was related to the floor and ceiling effects of the instruments. A useful regression equation was produced relating length of stay to Barthel score on admission. CONCLUSIONS: These data demonstrate the changes that occur during early inpatient rehabilitation after single incident brain injury. They explore clinical indicators of dependency and outcome, and aspects of resource utilization that characterize our service. Publication of data of this sort, from this and other units, should encourage the development and improvement of current rehabilitation service delivery after brain injury. PMID- 12745949 TI - Lower-extremity muscle forces measured by a hand-held dynamometer and the risk of falls among day-care users in Japan: using multinomial logistic regression analysis. AB - PURPOSE: To assess the relationship between the lower extremity muscle forces and risk of falls among the elderly who were using day-care services in the long-term care insurance system. METHOD: The study population comprised 96 women aged 69 - 91 years. At baseline, we assessed ADL, functional capacity, isometric muscle forces of knee extension and ankle dorsiflexion obtained with a hand-held dynamometer. After 1 year, we asked subjects about falls during the follow up period. RESULTS: In univariate analysis, only the knee extensor strength (p = 0.003) and ankle dorsiflexor strength (p = 0.02) differed significantly according to the fall status. After adjustment for age using multinomial logistic regression analysis, the weak quadriceps group was 7.50 times more likely than the strong group to fall once (p = 0.02), and 5.00 times more likely to fall twice or more (p = 0.02). The weakest ankle dorsiflexor group was 5.09 times more likely than the strongest group to fall twice or more (p = 0.05). CONCLUSIONS: Our results indicated that the forces of knee extensor and ankle dorsiflexor were strongly associated with falls among day-care users. Physiotherapy should focus on the lower-extremity muscle strength to prevent falls and deterioration of physical ability among elderly persons who are partially dependent and need support. PMID- 12745951 TI - The implementation of the 'International Classification of Functioning, Disability and Health' (ICF) in daily practice of neurorehabilitation: an interdisciplinary project at the Kantonsspital of Lucerne, Switzerland. AB - PURPOSE: The current paper describes the implementation of ICF as a standard language and framework for description of human functioning and disability for common use in every day work by the multiprofessional team. METHOD: An interdisciplinary project team involving all rehabilitation specialities was constituted. The extensive original document of ICF was broken down to a simplified raster for body functions and structures, activities and participation, as well as for contextual factors. These rasters had to cover the most important aspects concerning the patients treated on our unit. Checklists on the basis of these rasters were worked out for use by the different specialized teams. Using these checklists, rehabilitation conferences, form and language of interdisciplinary communication, goal setting and documentation were introduced newly in every day work for the interdisciplinary rehabilitation team, structured strictly based on the ICF-criteria. RESULTS: Since April 2002 the ICF-based processes are implemented in routine work for all members of the rehabilitation staff. First experiences show good acceptance by the team members, improvements in communication and documentation as well as substantial gains in content and handling of rehabilitation conferences. As a result of the implementation we observed, that participation, context and domiciliary interventions gained quite more influence in every day work at the unit. CONCLUSION: Implementation improved considerably the quality of interdisciplinary work processes and contributed to a more systematic approach to rehabilitation tasks by the team members. PMID- 12745952 TI - The impact of Internet access for people with spinal cord injuries: a descriptive analysis of a pilot study. AB - PURPOSE: A pilot study was undertaken regarding the effect of Internet access on health-related quality of life (HRQoL) and self-reported impact for people with spinal cord injuries (SCI). This study is unique in providing free Internet access and looking at benefits for people with disabilities. METHOD: HRQoL was measured using indicators of global health and social isolation at baseline and after 6 to 19 months of use via telephone interviews on a convenience sample of 23 adults with SCI. Additionally, monthly telephone surveys measured usage patterns, recreation, and self-reported impact. Both quantitative and qualitative analyses were conducted. Bivariate tests for differences in proportions and paired T-tests were conducted. Qualitatively, conceptual categories of impact were created using the Constant Comparative Method. RESULTS: Qualitatively, the predominant benefit was quality of life, mentioned by 61% of participants 46% of months surveyed, with quantitative trends towards improved emotional health. Ease of access to information, social connection, and quality of information were also frequently reported, with modest support from quantitative data. CONCLUSIONS: The study's persuasive qualitative results suggest the Internet has particular benefit to people with disabilities and that rehabilitation goals should include leisure. Further scientific research is strongly warranted. PMID- 12745953 TI - An investigation into the physical accessibility to wheelchair bound students of an Institution of Higher Education in South Africa. AB - PURPOSE: A descriptive cross-sectional study was undertaken to establish the ease of accessibility to wheelchair bound students of the campus of a large institution of higher education in South Africa. Accessibility was defined not simply in terms of access to buildings, but also of the added time and distance traveled by wheelchair bound students on the campus. METHODS: Five different faculties were randomly selected and typical routes travelled by a first year student in each faculty established. The shortest possible distances were mapped out for each faculty and measured using a metre wheel. For those in wheelchairs, this distance included the distance to ramps and to lifts. Mean speeds were used to determine the time taken to travel the measured distances. A checklist was used to measure the physical accessibility of the 18 buildings concerned. RESULTS: The total mean distance traveled between lecture theatre changeover by wheelchair-bound students was 402 metres (range 278-689 m for the different faculties), which was a mean difference of 66 m (range 11-145 m) longer than for ambulant students. The mean time taken was 17 minutes (range 11-28 minutes), which was 11 minutes (range 8-19 minutes) longer than for ambulant students. The Faculty of Mechanical Engineering fared the worst, at 689 metres and 28 minutes between changeover. Two of the 18 buildings measured were fully accessible, while three were completely inaccessible. Inaccessible toilets were the most common problem. CONCLUSIONS: Wheelchair-bound students consistently had to travel further and for longer times between lecture theatres in all the faculties measured. The students would therefore be unable to reach their lectures within the 10 minute time allocated by the university. The inaccessibility of the buildings limits the full integration of wheelchair-bound students into campus life. Certain administrative changes might assist in improving the ease of accessibility. Architectural adaptations, although more costly, might also prove to be effective. PMID- 12745954 TI - Walking patterns of hip arthroplasty patients: some observations on the medio lateral excursions of the trunk. AB - PURPOSE: This study examined the angular gait kinematics of the trunk and the pelvis in the frontal plane and their amount of side-to-side asymmetry in patients after total hip replacement arthroplasty. METHOD: The angular gait kinematics of 12 male hip arthroplasty patients (53-70 years) were compared to ten age-matched and ten young (24-35 years) male control subjects. Average step times and medio-lateral oscillation amplitudes of the pelvic and thoracic recordings were calculated for each step. Between successive steps the asymmetry ratio was computed and the mean angle around which the side-to-side oscillations occurred was compared to the angle in a symmetrical standing trial. RESULTS: ANOVA indicated no significant side differences in relative step cycle durations. Patients and senior controls had significantly (p<0.01) less pelvis side-to-side displacements than the younger controls. No significant between-group differences could be detected for the average asymmetry ratio. However, during walking the patients showed a significantly (p<0.01) increased lateral thorax and pelvis deviation. CONCLUSIONS: Hip replacement patients' thoracic and pelvic position is characterized by a lateral shift throughout the gait cycle, while left and right symmetry of angular movements amplitudes remain at about the same value of unimpaired subjects. PMID- 12745955 TI - Impairment, disability, and life satisfaction in Parkinson's disease. AB - PURPOSE: To determine the correlation of motor and musculoskeletal impairments with disability scores and life satisfaction. Musculoskeletal impairments might be due to the impact of motor impairments, however some cases may have these impairments even before the disease is diagnosed. METHODS: 48 consecutive patients (32 male, 16 female) with Parkinson's disease with a mean age of 65.6, mean age of onset of the disease of 59.8, and Hoehn and Yahr staging system 1-4 participated. Unified Parkinson's disease rating scale, modified Schwab and England activities of daily living scale and life satisfaction index were used. Also New York modified mobility rating scale for c-spine and upper extremities and trunk and lower extremities was performed. RESULTS: Motor and musculoskeletal impairments were correlated with the disability score. Bradykinesia, and tremor were correlated with upper and lower extremity range of motion limitation, but rigidity was not. Bradykinesia was correlated with life satisfication. Having had the disease longer was related to upper extremity range of motion limitation. CONCLUSIONS: Motor and musculoskeletal impairments are correlated with disability scores. Bradykinesia and musculoskeletal impairments are correlated with life satisfication. Patients with Parkinson's disease might benefit from a rehabilitation programme focusing on motor impairments such as bradykinesia as well as musculoskeletal impairments even in the early stages of the disease. PMID- 12745956 TI - Paraplegia-related alterations of bone density in forearm and hip in Greek patients after spinal cord injury. AB - PURPOSE: Paraplegia due to spinal cord injury is related with sublesional bone demineralization with an increased incidence of pathologic fractures in lower extremities. This study was carried out in order to evaluate bone density alterations in forearm and hip in Greek paraplegic patients after spinal cord injury and to correlate the findings with the level of injury, the neurological status, the time interval from injury and the performing of physiotherapy and therapeutic standing. METHOD: Fifty-seven paraplegic patients (33 men and 24 women, with injuries sustained from 6 months to 27 years) and 36 able-bodied age matched controls (25 men, 16 women) participated in the study. Bone mineral density (BMD) was measured by dual X-ray absorptiometry (DXA) in the proximal and distal forearm, the femoral neck, the greater trochanter and Ward's triangle. RESULTS: The measurements revealed a significant reduction of BMD of femoral neck (p<0.001 in male, p<0.001 in female paraplegics), greater trochanter (p<0.001 and p=0.001, respectively) and Ward's triangle (p=0.001 and p=0.005, respectively). Proximal forearm depicted non-significantly decreased BMD values and distal forearm depicted a slight increase in BMD values. The degree of demineralization was independent of factors such as complete or incomplete spinal cord injury, level of the lesion, physiotherapy and performing of standing. In addition to that, BMD values in both hip and forearm showed no statistically significant correlation with time after injury. CONCLUSIONS: BMD measurements in Greek paraplegic patients reveal bone loss, which most dramatically occurs in the region of hip with a consequent increase of fracture risk. Forearm measurements depict a non-homogeneous response with limited proximal bone loss and slight distal increase of BMD, the latter being possibly attributed to daily activities. PMID- 12745957 TI - Lifetime physical and sexual abuse in chronic pain patients: psychosocial correlates and treatment outcomes. AB - PURPOSE: This study describes a subgroup of diagnostically heterogeneous chronic pain patients, with a lifetime history of physical and/or sexual abuse, who underwent a pain management programme. A battery of psychosocial and pain measures were assessed, as well as 1-year post-treatment socio economic outcomes. METHOD: The prevalence of a history of abuse was assessed via a semi-structured interview of 162 consecutive patients (112 females and 50 males) presenting for 4 8 weeks of treatment in an interdisciplinary, outpatient rehabilitation programme. Treatment outcome data were gathered immediately, 6 months and 1 year following discharge. The chronic pain patients with a history of abuse were compared to those without a history of abuse on several pre-treatment psychosocial variables--pain severity, psychological distress, DSM-IV Axis I comorbidity and health care utilization. Patient groups were matched on age, race, primary pain diagnosis, time in pain prior to treatment and gender. RESULTS: Results indicated that 61% of patients had a history of lifetime physical and/or sexual abuse. Rates of sexual, and combined sexual and physical, abuse across the lifespan were higher for women than for men. Abused patients had a greater number of psychiatric diagnoses than nonabused patients. Abused patients also reported greater affective distress, less perceived life control, and a greater number of ER visits in the 6 months prior to treatment than their nonabused counterparts. A model consisting of gender (female), a higher number of psychiatric diagnoses, and higher affective distress was found to be a sensitive and relatively accurate predictor of abuse history. Finally, analyses indicated that, despite having greater psychosocial risk factors during the pre-treatment period, chronic pain patients with a history of abuse benefited from treatment and maintained treatment gains to a degree similar to nonabused chronic pain patients. CONCLUSIONS: Chronic patients with an abuse history can successfully complete a rehabilitation programme if the programme is designed to treat their psychosocial distress. Moreover, this also carries over to treatment outcome. A history of abuse does not have to negatively impact long-term treatment outcomes in this population of chronic pain patients. PMID- 12745958 TI - Satisfaction with rollators among community-living users: a follow-up study. AB - PURPOSE: Rollators are used in order to make mobility possible for people with restricted walking ability. The use of rollators is increasing, but little is known about outcomes. The aim of this study was to investigate users' satisfaction with rollators. METHOD: A follow-up study was carried out in seven Danish municipalities. One month after they got their device, 89 users of rollators were interviewed by means of the QUEST 1.0. Three months after the first interview a second interview took place and data from the 64 users available for follow-up were analysed. RESULTS: The users were satisfied with their rollators, and the frequency of use was high. However, many of the users were frail, and some of them were not fully satisfied in all respects. Women especially, users living alone and first time users were likely to be dissatisfied. The main problem identified was handling the rollator, and for several users the physical environment caused accessibility problems. CONCLUSIONS: Rollators are valuable for the users and a relevant societal intervention. However, a better match between person and technology, enhanced user training and follow-up can improve the outcome of the intervention. Furthermore, better rollator design is called for, and buses and the outdoor environment need to be made more accessible. PMID- 12745959 TI - An exploratory examination of an academic PM&R inpatient consultation service. AB - PURPOSE: This study sought to examine an inpatient consultation service delivery system at an academic teaching hospital. METHOD: Descriptive; retrospective; exploratory. Data from a 33 month period were analysed. Demographic profiles of patients receiving consultation were examined. A comparison was also made between alternate methods of delivering physiatric consultation. RESULTS: Only 80% of patients admitted to a teaching hospital during the study period received consultation. Referrals increased by 75% with the institution of a full-time consultation practice model. CONCLUSIONS: The utilization of an inpatient consultation service appears to be dependent upon the service delivery format. Further studies are needed to understand referral patterns and specific challenges to consultation services in an academic setting. Future research should focus on comparing clinical outcomes for patients in diagnostic categories who do and do not receive physiatric consultation. PMID- 12745960 TI - Evaluation and application of the General Motor Function assessment scale in geriatric rehabilitation. AB - PURPOSE: To test General Motor Function assessment scale (GMF) for concurrent validity, to analyse the distribution of scores and its clinical sensitivity in a setting of geriatric rehabilitation. METHOD: A descriptive and comparative study involving comparisons between GMF and Katz Index of ADL, analyses of GMF scores concerning distribution and comparisons between three geriatric care levels and between assessments pre- and post-intervention. One group of 20 in-patients and one group of 154 patients in three different care forms - institutional care, home rehabilitation and day care were included. Non parametric statistics were utilized, including a method which can separately measure the level of change for the group and for the individuals in that group. RESULTS: High correlation with r(s) = 0.80 (p < 0.001) between the subscale Dependence and the Katz Index of ADL verified that this part of the GMF measures variables related to ability in ADL. No floor effects in any of the subscales of the GMF were shown. The GMF was found to be sufficiently sensitive to demonstrate change from pre- and post intervention assessments. CONCLUSION: This study indicates that GMF could be useful as a satisfactorily valid and sensitive tool for physiotherapists' standardized assessments in geriatric practice. PMID- 12745961 TI - The Canadian Occupational Performance Measure as an outcome measure and team tool in a day treatment programme. AB - PURPOSE: To investigate the usefulness of the Canadian Occupational Performance Measure (COPM) in a day treatment programme for clients with rheumatoid arthritis. METHOD: The study was conducted in two parts. In the first part rehabilitation without changes in the programme was performed (n = 16). After that the COPM was introduced to all team members. In part two the COPM was used (n = 40). Clients' experiences of participation in the process were studied via a structured interview 2 - 4 weeks after discharge in both parts. Qualitative interviews were conducted with team members before part one and after completion of part two. RESULTS: Staff expressed that the COPM improved client participation in the rehabilitation process. Goals were formulated distinctly, and focused on activity and performance rather than function. Team conferences were focused on the client's needs. Outcome was considered clear and evident to the client. The changes in client routines demands thorough introduction, support and involvement, and takes time. Involvement and motivation for changing practice were difficult to obtain, this could be a result of a large staff turnover during the data collection period. CONCLUSIONS: The COPM should be seen as an aid to ensuring client participation in the goal formulation process, and facilitating treatment planning and evaluation of outcome. PMID- 12745962 TI - Quality of life after hip fracture: a comparison of four health status measures in 208 patients. AB - OBJECTIVES: We compared four health status measures for the evaluation of quality of life after hip fracture. METHODS: Two hundred and eight elderly hip fracture patients were followed up to 4 months after hospital admission. We used two interviewer-administered instruments (the Rehabilitation Activities Profile (RAP) and the Barthel Index (BI)) that focus on functional status, and two self assessment instruments (the Nottingham Health Profile (NHP) and the COOP/WONCA charts) that additionally include psychological and social health domains. The score distribution, internal consistency, construct validity, and sensitivity to change were investigated. RESULTS: At 4 months only 18% of surviving patients had reached the same level of functioning as before the fracture and, compared with reference values, lower scores of health status were found in the areas of physical mobility and emotional reactions. The number of comorbidities at hospital admission was the most important prognostic factor for recovery of health status at 4 months. The RAP and the BI both performed well in the assessment of functional status in regard to score distribution, internal consistency and construct validity. In contrast to the BI, the RAP also assessed instrumental activities of daily living and perceived problems with existing disabilities. The generic health status measures produced no added value in the assessment of functional status. The NHP covered a wider range of psychological health dimensions (emotion, pain, energy, and sleep) and had better psychometric properties than COOP/WONCA. None of the four instruments performed well in assessing social functioning. CONCLUSIONS: To assess health status after hip fracture, we recommend the RAP for functional status and the NHP for changes in emotion, pain, and energy. These instruments detected poor recovery in functional and emotional status at 4 months after fracture. PMID- 12745963 TI - A protocol for assessment of risk in wheelchair driving within a healthcare facility. AB - PURPOSE: The purpose of this article is to describe how a skilled nursing facility with significant wheelchair traffic developed a protocol for the multidisciplinary assessment and intervention management of patients whose wheelchair use represents a risk to self and others. This article describes the procedure and two case studies illustrating the utility of the protocol. METHOD: Staff of the facility developed a protocol for responding to sentinel events related to patients hurting themselves and others while using a wheelchair or observed using the wheelchair in an unsafe manner. The goal of the protocol is to determine the cause of the event, identify the necessary multidisciplinary interventions, and the evaluation of the effectiveness of the interventions. RESULTS: Case study analysis of the interventions indicated that a multidisciplinary evaluation and treatment plan leads to treatment interventions that suit the patient's medical condition while affording the patient the greatest amount of independence. CONCLUSION: Many patients who live in a long term skilled nursing facility must use either a manual or a power wheelchair for mobility. For these patients, the wheelchair may represent one of the last forms of independence they enjoy. The implementation of the protocol helped staff respond to important changes to the patient's condition and prepare a coordinated intervention. The patients found the protocol to be a helpful component of their treatment. PMID- 12745964 TI - The views of people with a physical disability on day activity centres in the Eastern Region of Ireland. AB - PURPOSE: The overall aim of this study was to evaluate the effectiveness of day activity centres in providing a service to physically disabled clients in the Eastern Region of Ireland. METHOD: Qualitative and quantitative methodologies were used. Four focus groups with clients were undertaken. This informed questionnaire content and design for subsequent quantitative work. Interviewer administered questionnaires were distributed to 156 randomly selected clients. Areas covered in the questionnaire included: demographic characteristics, income, living circumstances, medical information, access to day centre, activities at the centre, staff, outcome of attendance and satisfaction with the centre. RESULTS: There was a very positive response to the survey, all clients selected agreed to participate. The majority of clients were unmarried, unemployed and disabled due to a neurological disorder. While most lived in owner-occupied occupation, they were dependent on state benefits for their principal source of income. The majority of clients attended the centres for social and recreational reasons. Although there was a high level of satisfaction with the service overall, some aspects, such as availability of therapy and lack of continuity of staff are areas identified that need improvement. CONCLUSION: This study highlighted aspects of day activity services that clients value. The results of this evaluation should guide the proposed future development and expansion of this service. PMID- 12745965 TI - Designing health plan contracts for people with disabilities. AB - PURPOSE: In the USA, private businesses and the federal government contract with health plan companies to arrange and provide for specific health benefits and services. Most contracts are designed around acute care services for people without disabilities. We wanted to design a health benefit programme tailored to the needs of people with disabilities. METHOD: We convened two expert panels. The first, comprised of experts on government programme payments and benefit design, identified key services of a programme providing care for people with disabilities. The second panel, comprised of representatives of health plan companies, simulated the contract evaluation process. RESULTS: Model health benefit programmes for people with disabilities are similar to programmes for people without disabilities, but should provide and pay for care co-ordination, non-medical benefits, and unlimited therapy while incorporating self-directed care. As existing health plan companies consider whether or not to sign a contract to provide these programmes, operational issues carry as much weight as financial factors. CONCLUSION: Purchasers seeking to contract with health plans to provide innovative programmes for people with disabilities must be willing to consider pilot projects, and to give the health plan companies projected utilization and cost data so they can predict their financial risk. PMID- 12745966 TI - Training in brain injury rehabilitation. AB - PURPOSE: To evaluate our current training programme for professionals involved in brain injury rehabilitation, which involves disability simulation, and to consider the ethics and consequences of such programmes. METHOD: Seventy-two professionals took part in a brain injury disability simulation exercise and completed questionnaires pre and post the workshop. RESULTS: Results suggest that participants have increased insight into the challenges faced by people with brain injuries. CONCLUSIONS: We believe that brain injury simulation exercises help professionals increase their understanding of the impairments and disabilities that may follow brain injury and can therefore form an important part of a training programme for rehabilitation staff. PMID- 12745967 TI - Accessible cell phone design: development and application of a needs analysis framework. AB - PURPOSE: This research describes the development and use of the Needs Analysis and Requirements Acquisition (NARA) framework to elicit and construct user requirements for the design of cell phones (which are a type of assistive technology) that are both usable and accessible to persons with disabilities. METHOD: Semi-structured interviews and a focus group were used to elicit information and a systematic approach was used to translation information into requirements (construct). Elicitation and construction are the first two stages of NARA. RESULTS: Requirements for general and feature-specific phone attributes were identified, and several requirements were found to match six of the seven universal design principles. CONCLUSIONS: The study demonstrated that NARA is both a straight-forward and cost-effective method to develop user requirements and can be used throughout the development cycle. PMID- 12745968 TI - Thermal stress and radiation protection principles. AB - Exposure to radiofrequency (RF) fields can occur in residential, occupational and medical settings. Since many technologies use RF fields, it is important to fully investigate their effects on the human body. Since the demonstrated effect of RF exposure is heating, it is important to critically evaluate studies of elevated temperature effects on the human body, from the cellular and tissue level to the whole body level, including potential effects on the susceptible groups such as the very young and the very old. WHO convened a Workshop in the Spring of 2002 on the subject of Adverse Temperature Levels in the Human. The goal of the workshop was to evaluate most recent data useful for the development of science-based RF exposure limits. This paper outlines radiation protection principles that underline such an evaluation. It discusses the quality of literature needed for sound scientific reviews, provides the hierarchy of scientific evidence used to establish effects, distinguish between biological effects and adverse health consequences and indicates how evidence is evaluated. In addition, criteria for determining the most sensitive effects, the value of an effect that has a dose response and methods of extrapolation are also described. Finally, the need to account for scientific uncertainty in the formulation of guidance on exposure is discussed. PMID- 12745969 TI - Cardiovascular responses to heat stress and their adverse consequences in healthy and vulnerable human populations. AB - This paper reviews the basic thermoregulatory physiology of healthy people in relation to hazards from external heat stress and internal heat loads generated by physical exercise or radiofrequency (RF) radiation. In addition, members of the population are identified who may be particularly vulnerable to the effects of heat stress. These data are examined in relation to current international guidance on occupational and public exposure to RF radiation. When body temperature rises, heat balance of the body is normally restored by increased blood flow to the skin and by sweating. These responses increase the work of the heart and cause loss of salt and water from the body. They impair working efficiency and can overload the heart and cause haemoconcentration, which can lead to coronary and cerebral thrombosis, particularly in elderly people with atheromatous arteries. These adverse effects of thermoregulatory adjustments occur with even mild heat loads and account for the great majority of heat related illness and death. They are, therefore, particularly relevant to determination of safe population exposures to additional sources of heat stress. It is concluded that exposure to RF levels currently recommended as safe for the general population, equivalent to heat loads of about one tenth basal metabolic rate, could continue to be regarded as trivial in this context, but that prolonged exposures of the general population to RF levels higher than that could not be regarded as safe in all circumstances. PMID- 12745970 TI - Carcinogenic effects of hyperthermia. AB - The purpose of this paper is to assess the evidence for and against the premise that hyperthermia is carcinogenic. The paper is one of several published in this issue of the International Journal of Hyperthermia on the subject of the health risks of hyperthermia. The motivation for this issue of the journal was the result of a World Health Organization workshop that dealt with this issue, as it relates to exposure of the population to RF fields. Since hyperthermia can be a natural consequence of such exposures, the health risks of hyperthermia are relevant in this context. Particularly in the case of carcinogenesis, it is necessary to provide a brief overview of the data that have been generated to examine the carcinogenic risks of RF exposure, so that these results can be compared with studies that have examined the carcinogenic risks of hyperthermia. For this reason, the paper is organized into three sections dealing with: (1) effects of heat on DNA damage/repair and mutations, (2) in vivo studies evaluating the carcinogenic potential of heat alone and combined with other carcinogens, and (3) in vivo studies involving RF exposures. The bulk of the data presented indicate that hyperthermia alone is not carcinogenic. If hyperthermia occurs in the presence of exposure to known carcinogens, such as radiation or chemical carcinogens there is the potential for modulation of carcinogenic effects of those agents. In some circumstances, hyperthermia can actually protect against tumour formation. In other instances, hyperthermia clearly increases incidence of tumour formation, but this occurs following thermal exposures (several degrees C temperature rise for up to 1 h or more) and radiation (therapeutic levels as for treatment of cancer) or chemical carcinogen doses higher than would be encountered by the general population. The extrapolation of these results to the general population, where radiation exposure levels would be at background and temperature rise from incidental RF exposure, such as cell phones (which are estimated to cause no more than 0.1 degrees C temperature rise) is not recommended. Current evidence indicates that the temperature elevations resulting from RF exposure are not carcinogenic. Caution should be used in situations where exposure to known carcinogens is combined with thermal exposures high enough to cause tissue damage. A summary of thermal thresholds for tissue damage from hyperthermia is presented in another paper in this special issue (Dewhirst et al.). No data exist that examine the carcinogenic risks of chronic thermal exposures below the threshold for detectable tissue damage, either alone or in combination with known carcinogens. This is an important goal for future research. PMID- 12745971 TI - Cellular effects of hyperthermia: relevance to the minimum dose for thermal damage. AB - The specific mechanism of cell killing by hyperthermia is unknown, but the high activation energy of cell killing and other responses to hyperthermia suggest that protein denaturation is the rate-limiting step. Protein denaturation can be directly monitored by differential scanning calorimetry and in general there is a good correlation between protein denaturation and cellular response. Approximately 5% denaturation is necessary for detectable killing. Protein denaturation leads to the aggregation of both denatured and native protein with multiple effects on cellular function. PMID- 12745972 TI - Basic principles of thermal dosimetry and thermal thresholds for tissue damage from hyperthermia. AB - This paper is one of several in this Special Issue of the International Journal of Hyperthermia that discusses the current state of knowledge about the human health risks of hyperthermia. This special issue emanated from a workshop sponsored by the World Health Organization in the Spring of 2002 on this topic. It is anticipated that these papers will help to establish guidelines for human exposure to conditions leading to hyperthermia. This comprehensive review of the literature makes it clear that much more work needs to be done to clarify what the thresholds for thermal damage are in humans. This review summarizes the basic principles that govern the relationships between thermal exposure (temperature and time of exposure) and thermal damage, with an emphasis on normal tissue effects. Methods for converting one time-temperature combination to a time at a standardized temperature are provided as well as a detailed discussion about the underlying assumptions that go into these calculations. There are few in vivo papers examining the type and extent of damage that occurs in the lower temperature range for hyperthermic exposures (e.g. 39-42 degrees C). Therefore, it is clear that estimation of thermal dose to effect at these thermal exposures is less precise in that temperature range. In addition, there are virtually no data that directly relate to the thermal sensitivity of human tissues. Thus, establishment of guidelines for human exposure based on the data provided must be done with significant caution. There is detailed review and presentation of thermal thresholds for tissue damage (based on what is detectable in vivo). The data are normalized using thermal dosimetric concepts. Tables are included in an Appendix Database which compile published data for thresholds of thermal damage in a variety of tissues and species. This database is available by request (contact MWD or PJH), but not included in this manuscript for brevity. All of the studies reported are for single acute thermal exposures. Except for brain function and physiology (as detailed in this issue by Sharma et al) one notes the critical lack of publications examining effects of chronic thermal exposures as might be encountered in occupational hazards. This review also does not include information on the embryo, which is covered in detail elsewhere in this volume (see article by Edwards et al.) as well as in a recent review on this subject, which focuses on thermal dose. PMID- 12745973 TI - Effects of heat on embryos and foetuses. AB - OBJECTIVES: This paper reviews the effects of elevated maternal temperature on embryo and foetal development in experimental animals and in humans. CONCLUSIONS: Hyperthermia during pregnancy can cause embryonic death, abortion, growth retardation and developmental defects. Processes critical to embryonic development, such as cell proliferation, migration, differentiation and programmed cell death (apoptosis) are adversely affected by elevated maternal temperatures, showing some similarity to the effects of ionizing radiation. The development of the central nervous system is especially susceptible: a 2.5 degrees C elevation for 1 h during early neural tube closure in rats resulted in an increased incidence of cranio-facial defects, and a 'spike' temperature elevation of 2-2.5 degrees C in an exposure of 1 h during early neurogenesis in guinea pigs caused an increase in the incidence of microencephaly. However, in general, thresholds and dose-response relationships vary between species and even between different strains of the same species, depending on genotype. This precludes rigorous quantitative extrapolation to humans, although some general principles can be inferred. In humans, epidemiological studies suggest that an elevation of maternal body temperature by 2 degrees C for at least 24 h during fever can cause a range of developmental defects, but there is little information on thresholds for shorter exposures. Further experimental and epidemiological studies are recommended, focusing on stage-specific developmental effects in the central nervous system using a variety of sensitive assays. PMID- 12745974 TI - Hyperthermia induced pathophysiology of the central nervous system. AB - This review is focused on the pathophysiology of the central nervous system (CNS) associated with mild-to-moderate hyperthermia (body temperature > 37 degrees C but <40 degrees C) induced thermal stress in Human cases as well as whole body hyperthermia (WBH) in animal studies. Pathological changes can be observed in the nerve cells and glial cells in Humans following mild-to-moderate thermal exposure. On the other hand, morphological changes in the axons, nerve cells, glial cells and vascular endothelium is seen at the cellular and the molecular levels in rats subjected to heat exposure at 38 degrees C for 4 h (body temperature > 40 degrees C but <42 degrees C). This effect depends on the age of the animals and their prior thermal experiences. Taken together, heat stress induced hyperthermia, once believed to be non-toxic in the mammalian CNS, do produce specific alterations in the CNS that may have long-term behavioural, physiological and neuropathological consequences. The probable mechanism(s) underlying hyperthermia induced brain pathology is discussed. PMID- 12745975 TI - Effects of heat stress on cognitive performance: the current state of knowledge. AB - This paper discusses the current state of knowledge on the effects of heat stress on cognitive performance. Although substantial research has been performed, it has proven difficult to describe the literature findings in a systematic manner. This is due to the large number of factors that come into play, such as task type, exposure duration, skill and acclimatization level of the individual and due to the absence of a concise theory on which experimental work can be based. However, two trends have been identified. First, heat stress affects cognitive performance differentially, depending on the type of cognitive task. Secondly, it appears that a relationship can be established between the effects of heat stress and deep body temperature. A number of exposure limits have been proposed during the last decades. These limits are summarized in this paper, with a special emphasis on the most recent one derived by Hancock and Vasmatzidis. This limit, which employs an attentional resource approach, defines exposure duration thresholds as parallel lines. Although this approach appears to be the most promising thus far, it is concluded that much remains to be understood before a limit becomes universally acceptable. PMID- 12745976 TI - Summary, conclusions and recommendations: adverse temperature levels in the human body. AB - In the spring of 2002, The World Health Organization workshop 'Adverse Temperature Levels in the Human Body' brought together scientists with expertise in biological effects of hyperthermia to review the data and determine the evidence that could be used to evaluate potential adverse effects from human exposures to radiofrequency (RF) electromagnetic radiation in the range of 10-300 GHz. Standards for RF exposure in this frequency range are based currently on thermal effects. Information was reviewed on the ability of hyperthermia, either to the whole body or to part of the body to affect physiology, particularly the heart and circulatory system, to induce other thermoregulatory responses such as sweating, to affect the performance of simple and complex mental tasks, to induce various heat-related disorders such as heat stroke and to damage body tissue. Risks to a variety of organs were considered. In addition, thresholds for effects on developing embryos and foetuses and possible carcinogenic effects were also examined. These findings were discussed in the context of known cellular and biochemical responses of cells and tissues to hyperthermia. The experts judged the relevance of each study for informing decision-makers on the scientific basis for establishing safe exposure levels. The consensus was that standards should consider both temperature and time of exposure, whenever possible. PMID- 12745978 TI - Truck driver fatigue risk assessment and management: a multinational survey. AB - As part of an organizational review of safety, interviews and questionnaire surveys were performed on over 700 commercial goods drivers and their managers within a series of related companies operating across 17 countries. The results examine the reported incidence of fatigue-related problems in drivers and their associations with near miss and accident experience as well as work and organizational factors. Experience of fatigue problems while driving was linked to time of day and rotation of shifts, though most associations were small. There were significant associations found between fatigue experiences and driver and management systems of break taking and route scheduling. The quantitative combined with qualitative information suggested that, where feasible, more flexible approaches to managing the scheduling and sequencing of deliveries assisted drivers in managing their own fatigue problems through appropriate break taking. The results are interpreted within the overarching principles of risk assessment and risk control. PMID- 12745979 TI - Effects of wearing aircrew protective clothing on physiological and cognitive responses under various ambient conditions. AB - Heat stress can be a significant problem for pilots wearing protective clothing during flights, because they provide extra insulation which prevents evaporative heat loss. Heat stress can influence human cognitive activity, which might be critical in the flying situation, requiring efficient and error-free performance. This study investigated the effect of wearing protective clothing under various ambient conditions on physiological and cognitive performance. On several occasions, eight subjects were exposed for 3 h to three different environmental conditions; 0 degrees C at 80% RH, 23 degrees C at 63% RH and 40 degrees C at 19% RH. The subjects were equipped with thermistors, dressed as they normally do for flights (including helmet, two layers of underwear and an uninsulated survival suit). During three separate exposures the subjects carried out two cognitive performance tests (Vigilance test and DG test). Performance was scored as correct, incorrect, missed reaction and reaction time. Skin temperature, deep body temperature, heart rate, oxygen consumption, temperature and humidity inside the clothing, sweat loss, subjective sensation of temperature and thermal comfort were measured. Rises in rectal temperature, skin temperature, heart rate and body water loss indicated a high level of heat stress in the 40 degrees C ambient temperature condition in comparison with 0 degrees C and 23 degrees C. Performance of the DG test was unaffected by ambient temperature. However, the number of incorrect reactions in the Vigilance test was significantly higher at 40 degrees C than at 23 degrees C (p = 0.006) or 0 degrees C (p = 0.03). The effect on Vigilance performance correlated with changes in deep-body temperature, and this is in accordance with earlier studies that have demonstrated that cognitive performance is virtually unaffected unless environmental conditions are sufficient to change deep body temperature. PMID- 12745980 TI - Variability in mechanical exposure within and between individuals performing a highly constrained industrial work task. AB - Data on exposure variability is an important remedy for designing and interpreting studies of occupational ergonomics. The present study aimed at retrieving the within- and between-subjects variance of several mechanical exposure parameters in a repeated, short-cycle task. Seven experienced operators repeatedly secured joints using two types of hand-held nutrunners. The joints were placed in three different locations on a rack, simulating automotive assembly. Bilateral muscle activity from the upper trapezius and the lower arm extensors, as well as head and upper arm inclination was continuously monitored. Exposure levels and their variance components were assessed in several data subsets using ANOVA. The results were interpreted in terms of statistical precision and power, and discussed as markers of important ergonomic qualities. A substantial exposure variability was found within and between subjects in all joint locations and for both tools. For mixed work across tools and locations, the necessary number of subjects to arrive at a group mean exposure with 95% confidence limits corresponding to +/- 10% of the mean ranged between 8 and 158, with posture recordings tending to require smaller populations than muscle activity recordings. Within-subject variance increased 2- to 37-fold, depending on exposure parameter, when work was 'enlarged' from securings with a specified location and tool to a mix of all locations and tools. Systematic differences between subjects in variability and responsiveness to 'work enlargement' indicated individualized motor control strategies. The results illustrate the importance of exposure variability data to the design of proper measurement strategies. They also suggest that the sizes of exposure variability per se can be interpreted as operational indices of what is thought to be important ergonomic risk indicators, such as the 'sameness' of repeated operations and the allowance for flexible working techniques. PMID- 12745981 TI - Applying image descriptors to the assessment of legibility in Chinese characters. AB - The current study derived seven descriptors from the pixel matrix of 5401 Chinese characters in the commonly used Ming, Kai, Li, and Hei fonts. A factor analysis was used to reduce the number of descriptors for a validation experiment. The three-factor solution (number of strokes, character height, and character width) explaining 89.54% of the variance was derived from 5401 characters in four fonts. A 4 x 2 x 2 x 2 factorial experiment using fonts, number of strokes, character height, and character width as the independent variables was conducted to evaluate the impact of each critical factor on the legibility of Chinese characters. Each tested character was chosen from the 500 most commonly used characters to minimize the effect of familiarity and ensure that all participants could recognize the character if it was legible. The 16 university students who participated in the experiment were asked to identify a Chinese character initially displayed on a PC screen at its minimum size and enlarged gradually until the participant could recognize it. The analysis of variance suggests that all the main effects of font, number of strokes, character height, and character width are significant. The legibility thresholds of the four type styles from the most to the least legible are in the Hei, Ming, Kai, and Li sequence. Number of strokes is the only significant factor in predicting the legibility threshold for each individual font. However, if predicting the legibility threshold across four fonts, character height is the other significant factor with about the same predictive power as number of strokes. The legible threshold is increased by the number of strokes and decreased by the character height and character width. PMID- 12745982 TI - Responding to a fire emergency in a virtual environment: different patterns of action for different situations. AB - This paper presents an experimental study of participants' response to the sudden appearance of a fire emergency in a virtual environment (VE) and of the adaptivity of their response pattern. A VE has been built in which participants meet two situations: first an explorative navigation and afterwards a hurried escape from the unexpected outbreak of fire. Fire intensity and participants' distance from the exit at the outbreak of fire have been varied as well, to create different degrees of danger and different degrees of difficulty in the task of leaving the premises. Participants' action has been collected automatically for quantitative analysis by registering each individual activation of the interaction devices (a triple button joystick). In addition, the movements in both virtual and real environment of additional groups of participants have been videorecorded for qualitative analysis. Results show that the appearance of the fire emergency triggers important changes in the way people move in the VE, and that such changes are all adaptive responses to an emergency situation. In conclusion, people show recognition of a dangerous situation in a VE and readily produce adaptive responses, making the VE suitable for emergency simulations and for use as an effective training tool. PMID- 12745983 TI - How safe is cycling with a schoolbag? AB - This study was undertaken as a preliminary investigation to compare cycling errors and sudden stopping distance in secondary students traversing an obstacle course on their bicycle with and without a schoolbag, to determine if further studies of cycling accident aetiology amongst children were warranted. Twenty children cycled as fast as they could around an obstacle course. Each child undertook two separate trials, about an hour apart on a morning of the same day. The children traversed the course one at a time in turn using their own bicycle and wearing their own usual school clothes. For the first trial they did not carry a schoolbag. On the second trial they carried a schoolbag weighing 6 kg. There were no significant differences in the number of cycling errors made by the children when traversing the obstacle course with and without a schoolbag. However, the children generally took longer to complete the course when carrying the schoolbag and stopping distances were greater. It was not possible to be certain that these findings were genuine or whether the effect of carrying the schoolbag was reduced by increased familiarity with the course, since all of the children first traversed the course without a schoolbag and then with one. It is concluded that there is some justification for conducting further studies to explore the implications of carrying schoolbags on cycling accident aetiology amongst children. PMID- 12745987 TI - Effects of brevetoxins on murine myeloma SP2/O cells: aberrant cellular division. AB - Massive deaths of manatees (Trichechus manatus latirostris) during the red tide seasons have been attributed to brevetoxins produced by the dinoflagellate Karenia brevis (formerly Ptychodiscus breve and Gymnodinium breve). Although these toxins have been found in macrophages and lymphocytes in the lung, liver, and secondary lymphoid tissues of these animals, the molecular mechanisms of brevetoxicosis have not yet been identified. To investigate the effects of brevetoxins on immune cells, a murine myeloma cell line (SP2/O) was used as a model for in vitro studies. By adding brevetoxins to cultures of the SP2/O cells at concentrations ranging from 20 to 600 ng/ml, an apparent increase in proliferation was observed at around 2 hours post challenge as compared to the unchallenged cell cultures. This was followed by a drop in cell number at around 3 hours, suggesting an aberrant effect of brevetoxins on cellular division, the cells generated at 2 hours being apparently short-lived. In situ immunochemical staining of the SP2/O cells at 1 and 2 hour post challenge showed an accumulation of the toxins in the nucleus. A 21-kDa protein was subsequently isolated from the SP2/O cells as having brevetoxin-binding properties, and immunologically identified as p21, a nuclear factor known to down-regulate cellular proliferation through inhibition of cyclin-dependent kinases. These data are the first on a possible effect of brevetoxins on the cell cycle via binding to p21, a phenomenon that needs to be further investigated and validated in normal immune cells. PMID- 12745988 TI - Fire ant venom alkaloid, isosolenopsin A, a potent and selective inhibitor of neuronal nitric oxide synthase. AB - Massive, multiple fire ant, Solenopsis invicta, stings are often treated aggressively, particularly in the elderly, despite limited evidence of systemic toxicity due to the venom. Over 95% of the S. invicta venom is composed of piperidine alkaloid components, whose toxicity, if any, is unknown. To assess a possible pharmacological basis for systemic toxicity, an alkaloid-rich, protein free methanol extract of the venom from whole ants was assayed for inhibitory activity on the following nitric oxide synthase (NOS) isoforms, rat cerebellar neuronal (nNOS), bovine recombinant endothelial (eNOS), and murine recombinant immunologic (iNOS). Cytosolic NOS activity was determined by measuring the conversion of [(3)H]arginine to [(3)H]citrulline in vitro. Rat nNOS activity was inhibited significantly and in a concentration-dependent manner by the alkaloid rich venom extract. For nNOS, enzyme activity was inhibited by approximately 50% with 0.33 +/- 0.06 microg of this venom extract, and over 95% inhibition of the three isoforms, nNOS, eNOS, and iNOS, was found with doses of 60 microg in 60 microl reaction mixture. These results indicate that the alkaloid components of S. invicta venom can produce potent inhibition of all three major NOS isoforms. Isosolenopsin A (cis-2-methyl-6-undecylpiperidine), a naturally occurring fire ant piperidine alkaloid, was synthesized and tested for inhibitory activity against the three NOS isoforms. Enzyme activities for nNOS and eNOS were over 95% inhibited with 1000 microM of isosolenopsin A, whereas the activity of iNOS was inhibited by only about 20% at the same concentration. The IC(50) for each of three NOS isoforms was approximately 18 +/- 3.9 microM for nNOS, 156 +/- 10 microM for eNOS, and >1000 microM for iNOS, respectively. Kinetic studies showed isosolenopsin A inhibition to be noncompetitive with L-arginine (K(i) = 19 +/- 2 microM). The potency of isosolenopsin A as an inhibitor of nNOS compares favorably with the inhibitory potency of widely used nNOS inhibitors. Inhibition of NOS isoforms by isosolenopsin A and structurally similar compounds may have toxicological significance with respect to adverse reactions to fire ant stings. PMID- 12745989 TI - Changes in cross-fostered Sprague-Dawley rat litters exposed to perchlorate. AB - Ammonium perchlorate is used as an oxidizer in rocket fuel. It has become a groundwater contaminant, dissociating to ammonium cation and perchlorate anion. The perchlorate ion competes with iodide for uptake into the thyroid, reducing thyroid hormone production. Pregnant Sprague-Dawley rats were given either untreated or perchlorate (1 mg/kg-day) treated drinking water beginning on gestation day 2. One set of control and exposed dams was sacrificed on gestation day 20. The litters from the second set of control and exposed dams were crossed immediately after parturition and were sacrificed at postnatal day 10. Dam serum and thyroid, pooled fetal sera, and male and female pup sera were collected and analyzed for perchlorate, thyroid-stimulating hormone (TSH), triiodothyronine (T(3)), and thyroxine (T(4)). Control pups receiving perchlorate through lactation had serum levels at postnatal day 10 of 0.54 microg/ml and 0.56 microg/ml for male and female pups, respectively, whereas exposed fetuses had serum perchlorate levels of 0.38 +/- 0.04 microg/ml. Female pups receiving perchlorate lactationally had significantly lower levels of serum T(4) than control pups and prenatally exposed pups. Serum T(4) levels in male pups were not affected by perchlorate. Serum thyroid hormone levels from gestational perchlorate exposure were restored to control values by postnatal day 10. In utero perchlorate-exposure decreased serum T(4) levels in the fetus. Gestational studies in conjunction with a cross-fostering study design helped discern thyroid hormonal changes caused by perchlorate exposure during the perinatal period. PMID- 12745990 TI - Effects of lead on Na(+)-K(+) ATPase and Ca(+2) ATPase activities and lipid peroxidation in blood of workers. AB - Lead is considered one of the major environmental toxicants that causes hematological, neurological, and gastrointestinal dysfunction. In this study, the authors examined the relationship between lead and lipid peroxidation, lead and Na(+)-K(+) ATPase activity, and lead and Ca(+2) ATPase activity in blood of workers. The working group consisted of 30 male workers occupationally exposed to lead at least for 10 years. The control group consisted of 20 healthy male individuals not involved with job-related lead exposures. Blood lead content of the control group and the working group were 10.0 +/- 1.8 microg/dl and 317.3 +/- 47.6 microg/dl, respectively. Malondialdehyde (MDA) value of the working group (0.57 +/- 0.30 nmol MDA/ml) was significantly greater than MDA value of the control goup (0.17 +/- 0.02 nmol MDA/ml). In the working group, both Na(+)-K(+) ATPase activity (105.0 +/- 47.0 nmol Pi. mg protein(-1) x h(-1)) and Ca(+2) ATPase activity (58.0 +/- 40.0 nmol Pi. mg protein(-1) x h(-1)) were lower compared with the corresponding values of Na(+)-K(+) ATPase activity (247.0 +/- 41.0 nmol Pi. mg protein(-1) x h(-1)) and Ca(+2) ATPase activity (230.0 +/- 41.0 nmol Pi. mg protein(-1) x h(-1)) of normal controls. The results show that lead exposure causes inhibition of Na(+)-K(+) ATPase and Ca(+2) ATPase activities and also results in increased lipid peroxidation. PMID- 12745991 TI - Comparison of in vivo (Draize method) and in vitro (Corrositex assay) dermal corrosion values for selected industrial chemicals. AB - Skin irritation is a common occupational hazard for employees engaged in the manufacture, transport, and use of industrial chemicals. The most common method used to evaluate dermal irritation and/or corrosion has typically been in vivo tests using rabbits (Draize method). Several in vitro test methods have been developed, with Corrositex being the first to gain approval by a regulatory agency (U.S. Department of Transportation). The purpose of this study was to compare the results of in vitro (Corrositex) assays of dermal irritation/corrosion to in vivo test data for several industrial chemical formulations and to determine the predictability and usefulness of the Corrositex assay for these types of products. Twenty-four (24) formulations were qualified, categorized, and evaluated using the Corrositex method and the results compared to available animal data for each of the formulations. The Corrositex assay accurately predicted a corrosive end point in 8 (57.1%) of the 14 formulations identified as corrosive by the in vivo evaluations. Corrositex accurately predicted a noncorrosive end point for 1 (10%) of 10 formulations determined to be noncorrosive in animal studies. The Corrositex assay overpredicted the packing group for 12 (50%) of the 24 formulations, and underpredicted the packing group for 7 (29.2%) of the 24 formulations. Compared to the in vivo results, Corrositex correctly classified as corrosive or noncorrosive 37.5% of the formulations tested. A concordance of 20.8% for the packing group assignments of the evaluated formulations was calculated. The Corrositex assay did not accurately predict a corrosive end point or packing group assignment for all of the formulations used in this study. Manufacturers should assess the relevance of this method to their products prior to relying on it for compliance with hazardous material and worker safety regulations. PMID- 12745992 TI - Widespread arsenic contamination of soils in residential areas and public spaces: an emerging regulatory or medical crisis? AB - A critical review finds government agencies allow, permit, license, or ignore arsenic releases to surface soils. Release rates are controlled or evaluated using risk-based soil contaminant numerical limits employing standardized risk algorithms, chemical-specific and default input values. United States arsenic residential soil limits, approximately 0.4- approximately 40 ppm, generally correspond to a one-in-one-million to a one-in-ten-thousand incremental cancer risk range via ingestion of or direct contact with contaminated residential soils. Background arsenic surface soil levels often exceed applicable limits. Arsenic releases to surface soils (via, e.g., air emissions, waste recycling, soil amendments, direct pesticide application, and chromated copper arsenic (CCA) treated wood) can result in greatly elevated arsenic levels, sometimes one to two orders of magnitude greater than applicable numerical limits. CCA-treated wood, a heavily used infrastructure material at residences and public spaces, can release sufficient arsenic to result in surface soil concentrations that exceed numerical limits by one or two orders of magnitude. Although significant exceedence of arsenic surface soil numerical limits would normally result in regulatory actions at industrial or hazardous waste sites, no such pattern is seen at residential and public spaces. Given the current risk assessment paradigm, measured or expected elevated surface soil arsenic levels at residential and public spaces suggest that a regulatory health crisis of sizeable magnitude is imminent. In contrast, available literature and a survey of government agencies conducted for this paper finds no verified cases of human morbidity or mortality resulting from exposure to elevated levels of arsenic in surface soils. This concomitance of an emerging regulatory health crisis in the absence of a medical crisis is arguably partly attributable to inadequate government and private party attention to the issue. PMID- 12745993 TI - Antibiotic resistance: who is winning the war? Introductory remarks. AB - The evolutionary response of bacteria, fungi, viruses, and parasites to the selective pressure exerted by antimicrobial agents is the emergence of populations that resist the action of the antimicrobial. The emergence and dissemination of such resistance in a variety of these pathogens is a growing public health concern. In response, the scientific community developed an action plan to address this public health issue. Antimicrobial, antifungal, and antiviral drug development for the treatment of diseases caused by resistant pathogens is one component of this strategy. In addition, due to the targeting of specific drugs against resistant pathogens, we may more readily accept a given drugs toxicity profile for the added therapeutic benefit. This symposium provides a discussion of the modes of action and mechanisms of resistance to antimicrobial agents, and the use of surveillance systems to help understand the nature and magnitude of resistance. The goal is to help guide antimicrobial drug product development and use. Specific toxicity issues are presented that should be considered in phase 1 development of antimicrobial drug products for use in clinical medicine and veterinary medicine. Finally, the national and global strategies developed by federal agencies in the Public Health Action Plan to Combat Antimicrobial Resistance are outlined. PMID- 12745994 TI - Animal drug human food safety toxicology and antimicrobial resistance--the square peg. AB - This paper presents the traditional approach for the evaluation of human food safety used for animal drugs intended for food animals, and describes some of the difficulties posed by antimicrobial drug resistance. Like human drugs, animal drugs must be safe and effective for the patient. However, unlike human drugs, food derived from animals treated with the animal drug must also be shown to be safe for human consumption. The Food and Drug Administration has come to realize that antimicrobial drugs used in the treatment of the food animal have the potential to create a unique residue--increased numbers of microorganism that are resistant to antimicrobial drug treatment. The traditional toxicological paradigm for chemical residues does not apply to this unique microbiological residue. Information useful to a food safety evaluation may include the potential for the animal antimicrobial drug to diminish the susceptibility of microorganisms to human antimicrobial drugs, any human medical use of the drug, relationship to other human antimicrobial drugs, and the ability of the animal drug to alter the susceptibility of relevant microorganism to important human antimicrobial drugs. Yet to be developed are standardized approaches to quantify an acceptable level of resistant microorganism in food and to mitigate the hazard to assure that there is a reasonable certainty of no harm following the consumption of the edible food derived from the treated animal. PMID- 12745995 TI - Antimicrobials: modes of action and mechanisms of resistance. AB - After six decades of widespread antibiotic use, bacterial pathogens of human and animal origin are becoming increasingly resistant to many antimicrobial agents. Antimicrobial resistance develops through a limited number of mechanisms: (a). permeability changes in the bacterial cell wall/membrane, which restrict antimicrobial access to target sites; (b). active efflux of the antimicrobial from the cell; (c). mutation in the target site; (d). enzymatic modification or degradation of the antimicrobial; and (e). acquisition of alternative metabolic pathways to those inhibited by the drug. Numerous bacterial antimicrobial resistance phenotypes result from the acquisition of external genes that may provide resistance to an entire class of antimicrobials. These genes are frequently associated with large transferable extrachromosomal DNA elements called plasmids, on which may be other mobile DNA elements such as transposons and integrons. An array of different resistance genes may accumulate on a single mobile element, presenting a situation in which multiple antibiotic resistance can be acquired via a single genetic event. The versatility of bacterial populations in adapting to toxic environments, along with their facility in exchanging DNA, signifies that antibiotic resistance is an inevitable biological phenomenon that will likely continue to be a chronic medical problem. Successful management of current antimicrobials, and the continued development of new ones, is vital to protecting human and animal health against bacterial pathogens. PMID- 12745998 TI - The study of comparative gerontology. PMID- 12746000 TI - Founding and processes of the International Council on Women's Health Issues: attentive partnering. The first 19 years. AB - I present a theoretical analysis and global influence of the first 19 years of the International Council on Women's Health Issues (ICOWHI). To do so, I analyzed observational, documented, and casual data using the constant comparative method of classical grounded theory. All data bits were analyzed with one another, coded, categorized, and reexamined until I discovered a central category that seemed to explain most of the social scene under study. The explanatory power of the central category, attentive partnering, is presented in terms of its applicability to the interactive relationship between participants working for a common goal, and within the cultural, temporal, social, and social structural context of the relationships. Partnering can occur wherever individuals with a common purpose assemble. The conditions under which the interactive process of attentive partnering occurs seem to be the presence of determined, persuasive leaders, who foster growth-enhancing collegial relationships. When this social system is in place, a small volunteer organization can have a global impact. PMID- 12746002 TI - Older women caring for spouses with Alzheimer's disease at home: making sense of the situation. AB - I combined phenomenological and feminist approaches to capture older women's own description of their Alzheimer's disease (AD) caregiving experience. Face-to-face interviews were conducted with 14 women who were 60 or older and cared at home for their spouses diagnosed with AD. Data were analyzed using a modified version of Colaizzi's method. Five major themes were found to define the fundamental structure of these women's experience. Here I address only findings pertaining to the fourth theme: making sense of the situation. These findings reveal aspects unique to older women's/wives' AD caregiving experience. PMID- 12746001 TI - The influence of income on the experience of informal caregiving: policy implications. AB - Examining the need for an eldercare policy is relevant and timely because the population is aging, a focus of care has shifted from institutional to community based, and informal caregivers, primarily women, are increasingly pressured to be responsible for eldercare. The purpose of the study is to examine the differences in the experiences of low-income and those who are not low-income informal caregivers. Three hundred questionnaires were mailed to past and present home care clients and 58 questionnaires were returned (19% response rate). This research revealed that low-income caregivers have increased needs for support and education from those who can afford to pay for their support services. Low-income caregivers experience significantly greater caregiver distress than do caregivers who are not low income. The unique needs of low-income caregivers must be considered in the formation of Canadian eldercare policy as increased health care privatization promotes the growing inequality in health care provision. PMID- 12746003 TI - Physical abuse in low-income women in Aleppo, Syria. AB - Violence against women is a vicious practice present in all societies. Yet data about its occurrence and associated factors are scarce in the Arab world. In this study, we attempt to determine the spread of physical abuse and its sociodemographic correlates among low-income women in Aleppo, Syria. A sample of 411 women was recruited from 8 randomly selected primary care centers in Aleppo. Response rate was 97%, mean age of participants 28 +/- 8 years, and most women (88%) were married. A special questionnaire was used including questions about physical abuse, the self-reporting questionnaire (SRQ-20), and questions about relevant sociodemographic information. Current physical abuse (battering at least 3 times during the previous year) was found in 23% of the investigated and among 26% of married women, while regular abuse (battering at least once weekly) was found in 3.3% of married women. Correlates of physical abuse were women's education, religion, age, marital status, economic status, mental distress, smoking, and residence. Our data show that physical abuse is prevalent in this population and that women's education is the most important modifiable factor. PMID- 12746004 TI - Depression during early recovery from heart surgery among early middle-age, midlife, and elderly women. AB - Theories and studies about the psychological processes of midlife propose a transition that may result in a more integrated personality structure throughout the second half of life. This more integrated structure includes concepts such as generativity, self-assertion, and independence. However, this structure, especially in early middle-age, can be affected by a life-threatening health disruption such as heart disease. One hundred and fifty-five women participated in a cross-sectional survey designed to investigate depression in early middle age, midlife, and elderly women who had undergone heart surgery. Women aged 40-55 years composed the early middle-age group, women aged 56-65 years composed the midlife age group, and women aged 66+ years were identified as the elderly age group. Affective components of depression were measured by asking participants to respond to three items regarding frequency of sadness, depression, and "the blues." Affective depression scores were summed and a mean score derived. Mean scores significantly differed by age group (F [2, 152] = 3.05, p =.05). Older women in the study fared better than their younger counterparts in terms of depression scores after a major cardiac health disruption. Post hoc comparisons indicated that mean scores for the early middle-age participants were significantly higher than for the midlife and older participants. Depression in the context of a cardiac health disruption appears to be linked to the developmental stage of a woman's life trajectory on which it is superimposed. PMID- 12746005 TI - The artistry and ability of traditional women healers. AB - In a phenomenological research study with a purposeful sample, 6 Ojibwa and Cree indigenous women healers from Canada and the United States shared their experience of being a traditional healer. Using stories obtained during open ended, unstructured interviews, in this article I depict the lives, backgrounds, and traditional healing practices of women who, in the past, have not been afforded an opportunity to dialogue about their healing art and abilities. The methods of these women healers, their arts and their gifts, are different from those of Western conventional medicine because of dissimilar world views related to health and illness. An increased awareness of health care providers related to the ancient art of traditional healing currently practiced in communities by gifted women who provide culturally specific holistic healing and health care is essential. PMID- 12746006 TI - Women with heart disease: can the common-sense model of illness help? AB - Noncompliance with cardiac rehabilitation programs is a major concern for female coronary heart disease patients. In this article I argue for the use of the common-sense model of illness in developing interventions to increase compliance with cardiac rehabilitation programs among women with heart disease. First, the common-sense model of illness is discussed. Second, a personal narrative technique that addresses the key components of the common-sense model of illness is presented. I recommend that a modified version of the personal narrative technique be used to increase women's compliance with cardiac rehabilitation programs because this technique is well suited for women's health issues. PMID- 12746007 TI - The U.S. health care system is dysfunctional. PMID- 12746009 TI - Questioning the construction of maternal age as a fertility problem. AB - The belief that fertility problems derive from maternal age, increasing markedly at 35, reflects social constructions of biology in developed nations. These constructions perpetuate a negative view of female aging. However, research since 1985 can be interpreted to suggest that there is no, or minimal, association between maternal age and problems associated with fertility. Differences in problems between pre- and postmaternal age 35 fertility can be explained by social conditions occurring with fertility, notably, parenting decisions, physical problems with the male and the potential child, and medical intervention. Once we look at fertility as a relationship among the woman, the man, and the potential child, rather than the woman only, we can see these factors. These components suggest that maternal age may be related only tangentially to successful fertility. PMID- 12746010 TI - The clientele of traditional birth homes in rural southeastern Nigeria. AB - Although it is widely documented that traditional birth homes (TBHs) do more than deliver babies, little is known about the other functions in addition to child delivery, which TBHs perform. Drawing on in-depth individual interviews with 13 traditional birth attendants (TBAs) and 147 users of TBHs, we profile the characteristics and health conditions of the clientele of TBHs in four rural communities in southeastern Nigeria. We found that TBHs provide their clients, who are mainly less educated women and girls, health services that range from child delivery, child sex selection, and abortion to family planning and cures for vaginal bleeding. Women are attracted to TBHs because the services are low cost, the women require privacy about their conditions, the TBHs are close by, and the women are confident in the abilities of TBHs. Rural women are bound by poverty, culture, and local values in their choices of services. We assert that health interventions to local people will need to be couched within frameworks that are responsive to their socioeconomic and cultural sensitivities if they are to deliver their expected impact. PMID- 12746011 TI - A process model of Depo-Provera use in Canadian women. AB - This study investigated women's experiences with the long-acting, injectable contraceptive Depo-Provera. Fourteen women were interviewed and 57 charts of clients at a Canadian Sexual Health Centre were reviewed to generate a theory of use. A three-stage process model describes women's choice of this contraceptive and decision to continue or discontinue use. This model suggests that women's use of Depo-Provera is determined by a complex interaction of individual cognition, relational processes, and sociocultural contexts. PMID- 12746012 TI - The impact of ectopic pregnancy: a 16-year follow-up study. AB - Ectopic pregnancy is a potentially life-threatening event that represents the loss of a pregnancy and also may have longer-term consequences for fertility. Despite this triple threat to a woman's well-being, almost no systematic research exists on the psychological impact of ectopic pregnancy. We sought to reinterview 18 women who had been interviewed 2 months following an ectopic pregnancy as part of a longitudinal study of pregnancy loss 16 years previously. Thirteen of these women responded to questions about the long-term impact of the ectopic pregnancy on their lives in the intervening years. For many of the women, the ectopic pregnancy was a traumatic experience that impaired their fertility, strained their marriages, and led to a crisis of faith. Yet, despite the trauma of the event, overall the women found ways to interpret it positively and to use it as a source of meaning in their lives. PMID- 12746013 TI - Influence of oral contraceptive use on pain perception and blood pressure. AB - The purpose of this study was to examine the influence of oral contraceptive (OC) use on pain perception and blood pressure. Thirty women (15 using OC and 15 normally menstruating) reported to the laboratory once a week for 4 consecutive weeks. Blood pressure and heart rate were assessed, and then pressure was applied to the left forefinger for 2 minutes with the Forgione-Barber pain stimulator while participants indicated when the stimulus became painful (PT) and rated the intensity of pain (PR). Data were analyzed with a 2 (group) x 4 (trials) ANOVA. Results indicated no significant differences (P > 0.05) for PT or PR between the two groups of women across the 4 weeks. There was a significant trials effect for systolic blood pressure (SBP; P < 0.05), with SBP being higher premenstrually and menstrually compared with postmenstrually in both groups of women. It is concluded that OC use did not significantly influence pain perception or blood pressure. PMID- 12746014 TI - Abortion--it is for some women only! Hmong women's perceptions of abortion. AB - In this article, I explore traditional Hmong explanations about abortion and the ethnomedical knowledge and practices that pertain to it. The article derived from in-depth interviews and participant observation with the Hmong who are now living in Melbourne, Australia. Hmong women are knowledgeable about indigenous fertility control methods including abortion. However, only women who are older and have had many children to ensure the continuity of the lineage have a right to abortion. Younger women do not have this right. Women may wish to control their fertility by means of abortion, but their wishes may be constrained by a societal norm that places a great deal of pressure on women to continue producing children. In a culture that values having many children, abortion is not easily accepted because it upsets the cosmological balance of the society. This has implications for younger Hmong women who may wish to control their fertility in Australia and elsewhere. PMID- 12746015 TI - Out of sync: a generation of first-time mothers over 30. AB - Increasing numbers of women have delayed childbearing until after they reach their thirties. The researchers who studied early maternal role attainment in women over 30 failed to reflect on the challenges of raising a child to adulthood. The use of grounded theory helped us understand the extended mothering experience from the perspective of women over 30. Fifty-three women from Indiana, Illinois, Michigan, California, Connecticut, and Florida composed the total sample. Ages of the women ranged from 30 to 91; ages of the children ranged from 6 weeks to late sixties. We called the dominant category out of sync, with properties that included mortality, vitality, and social structural considerations. The pervasive quality of the central variable makes this account of interest to an international audience. PMID- 12746016 TI - Measuring gynecological morbidity: evaluating two different data sources from Beirut. AB - The lack of consensus over the most appropriate source to use in assessing reproductive morbidity could, in part, explain the inadequacy of available information on the subject. To outline this situation, gynecological morbidity data collected from two different sources in Beirut, Lebanon, namely, population based health interviews (779 ever-married women aged between 15 and 49) and private gynecologists' clinics (867 women with similar characteristics), are described. Although neither source is likely to represent the true prevalence of gynecological conditions, both agree sufficiently to shed light on the importance of some conditions such as menstrual disturbances (15% in both samples), infections/inflammations (17% in the households sample), and infertility-related concerns (12% in the clinics sample). Interestingly, despite the demographic differences, the most common conditions that the women complained about and the most common diagnoses that the gynecologists offered were similar for both samples. Therefore, given that the logistics in the gynecologists' clinic survey were easier, we recommend the use of health service data in settings where a representative sample of providers can be identified and service use is high. PMID- 12746018 TI - Communication patterns between health care providers and their clients at an antenatal clinic in Zimbabwe. AB - We conducted this study to explore communication between pregnant women and health care providers at an antenatal clinic in Zimbabwe. We adopted a field research approach using qualitative strategies for data collection and analysis. Using content analysis, we identified impersonal communication, nonprivate communication, rigid communication, uninformed communication, and authoritative communication as characteristic components of the interaction between the pregnant women and the health care providers. The attitudes reflected by the communication styles at the antenatal clinic must be viewed in relationship to the culture and tradition of Zimbabwe. With further research, we may identify obstacles to communication that are not related to culture. This knowledge may lead to a better understanding of appropriate communication strategies to promote healthy pregnancy. PMID- 12746019 TI - Promoting attentional health: importance to women's lives. AB - Promoting attentional health is an important opportunity to support and empower women to actively participate in their own health care and function effectively in multiple roles. Attention, the ability to focus or concentrate, is a necessary cognitive resource used throughout the life span in participating in self-health care. However, women can inadvertently overuse this essential capacity, especially when responding to life's multiple demands, resulting in mental or attentional fatigue. When women experience this mental fatigue, many daily activities, including self-health care, are more difficult. An attentional perspective in women's health allows approaches to improve daily functioning and reduce the risk of attentional fatigue. In this article we present the conceptual dimensions of directed attention and its relevance to women's health; practice and research implications are identified. PMID- 12746020 TI - Making culture visible: an examination of birthplace and health status. AB - Culture shapes beliefs about health and illness. It provides the context by which an individual evaluates his or her symptoms to decide whether he or she is healthy or ill. We have largely ignored the impact of culture on the health status of U.S.- and foreign-born Black women. We have operationalized the term Black or African American in a way that obscures cultural differences among women of African descent living in the United States. While we know that people of African descent living in the United States are composed of many different ethnic backgrounds (e.g., persons from the Caribbean and African countries), we have not systematically examined these differences using national data. In this study I examine the relationship between culture and health status using the U.S. Department of Health and Human Services 1991 National Health Interview Survey (NHIS). Significant differences were found between U.S.- and foreign-born Black women. Foreign-born Black women reported more favorable health status than U.S. born Black women. Measures of socioeconomic status and social support accounted for many differences between the two groups. These findings suggest that culture may play a role in the evaluation of health status. Therefore, U.S.- and foreign born Black women appear to use different criteria when assessing their health status. PMID- 12746021 TI - The cultural context of gender, identity: female genital, excision and infibulation. AB - Our goal is to explore the practices of female genital excision and infibulation as they relate to gender identity and the acculturation process in Canada. We examined relevant research on these issues and share the results of a nationwide project conducted in 1997-1999 among 162 Canadian immigrants from regions in Africa where practices of excision and infibulation are still in effect. Our discussion of gender identity is inextricably linked to notions about the ways in which girls, women, and virginity are socially constructed. The complexity of the acculturation process along with the integration within a host society is highlighted and the conflicting identities available to women are brought to the fore. PMID- 12746022 TI - Barriers to wellness activities for Canadian women with physical disabilities. AB - Although there is a great deal of interest in women's health, research on the health and well being of women with disabilities has not increased. In this article we present internal and structural barriers to wellness activities experienced by women with disabilities. We also discuss women's actual and recommended strategies to address these barriers. Data were collected in six focus groups in urban and rural Ontario, Canada. The participants represented a diversity of disability, age, and ethnoracial backgrounds. Our findings suggest that individual and structural barriers exist for the women, with structural barriers (physical, informational, and systemic access) being predominant. Barriers prevented women from engaging in desired wellness activities. Women discussed actual strategies to address these barriers, such as collective efforts to buy nutritious foods and recommendations to create greater access (e.g., increase health professionals' training in disability issues). PMID- 12746023 TI - Preparedness of health care practitioners to screen women for domestic violence in a border community. AB - Health care professionals do not always screen patients for domestic abuse. In the absence of screening, the likelihood of misdiagnoses and prescriptions contraindicated for symptoms of abuse survivors is increased. Several factors may contribute to the lack of routine and uniform screening, including a lack of educational preparedness. The qualitative findings presented here are part of a larger quantitative study investigating the skills, beliefs, and expectations about screening for domestic abuse among physicians, dentists, and nurse practitioners from a border community in southwest Texas. Practitioners requested specific information to enhance educational preparedness and improve clinical practice for their abused patients. PMID- 12746024 TI - Attitudes and practices of doctors toward domestic violence victims in South Africa. AB - Our study intended to ascertain doctors' attitudes and practices toward domestic violence in a sample of 402 doctors randomly chosen for a mail survey from a list of registered medical practitioners in South Africa. We found that the mean number of patients treated for domestic violence per month was 11.4 (SD = 13.4); for White doctors 15.8, African Black 6.2, and for Asian doctors 5.3. The high frequency of domestic violence detected in this sample seems to reflect national surveys on domestic violence. Most doctors believed that they should play a role in prevention and treatment, and their attitudes toward victims of domestic violence were generally sympathetic and supportive. Male doctors and long-serving practitioners tended to have more conservative views of the causes and attitudes of spousal assault and of the doctors' role in prevention. Only 9.7% of the doctors had received any training on domestic violence. Further research is needed to establish the prevalence of domestic violence in women presenting to medical practitioners and to investigate how the problem currently is being addressed. PMID- 12746025 TI - Reflections on September 11, 2001. PMID- 12746027 TI - Getting no respect: barriers to mammography for a group of Swedish women. AB - Mammography screening is a highly sensitive and specific method to detect breast cancer at an early stage. If screening campaigns are to be cost effective, compliance is valuable. However, many women do not attend when called for mammography screening. Our aim in this study is to understand and explain why women become nonattenders. A sample of 16 nonattending women, aged between 43 and 73 years, participated in this qualitative study, by interviews or written comments. The core category discovered in the data was "getting no respect." The informants did not feel respected from either the society or the health care system. Below this core category, two categories were identified: the mammography examination and affecting circumstances. The performance and its effects comprised the content of the category of mammography examination. The category affecting circumstances included knowledge about risk factors, prevention, and practical or emotional arguments. PMID- 12746028 TI - Japanese American women: behaviors and attitudes toward breast cancer education and screening. AB - Moores University of California, San Diego, Cancer Center's Asian Grocery Store Based Cancer Education Program trained bilingual, bicultural student health educators to provide breast cancer information to Japanese American women. A subset consented to help evaluate the program by completing baseline and follow up surveys. Study participants reported high adherence to mammography screening guidelines, but lower than optimal adherence to clinical breast examination (CBE) and monthly breast self-examination (BSE) guidelines. While less than half of the women felt they had enough knowledge about breast cancer, nearly all indicated that they would be willing to share any knowledge they gained with loved ones and that their loved ones would be receptive to their information. A limitation of the study is its small sample. PMID- 12746029 TI - The meaning of health in mammography screening for African American women. AB - Inadequate use of mammography screening for early detection of breast cancer is an important factor associated with the disproportionate breast cancer death rates in African American women. To improve understanding of the mammography screening experience and health for African Americans, focus groups were held with 30 African American women. Seven categories emerged: (a) the mind, body, and spirit connection: (b) living your life; (c) looking good; (d) good health-bad health; (e) prevention-detection confusion; (f) being afraid of cancer; and (g) what gets in the way. Implications for developing tailored messages and for addressing system barriers are discussed. PMID- 12746030 TI - Women's views of a breast screening service. AB - We performed this study to better understand women's views of the breast screening experience in order to improve the rate of uptake and the service offered. The sample comprised 27 women aged over 50 who had attended a regional breast screening service and received normal results. Data were collected through four focus groups. Results showed a need for a local, easily accessible breast screening unit with free car park facilities. Thus, women preferred a mobile screening unit that had a "cosy, nonclinical" atmosphere. Staff were reported to be efficient, caring, and helpful. Nonetheless, all women reported discomfort during the mammogram, and the nature of the procedure, during which women had their breasts squashed between two plates, was unexpected. Furthermore, women expressed anxiety about results, receiving recall letters, and about cancer in general. Women suggested the service should be advertised more widely and wanted more detailed information before and during screening. They demonstrated altruistic concern for other women who are currently excluded from the screening programme. PMID- 12746031 TI - Traditional breastfeeding practices of the Ojibwe of Northern Minnesota. AB - The Ojibwe have transitioned over the past 100 years from a woodland people moving with the seasons, to forced confinement on rural reservations, to inner city poverty. Traditionally, Ojibwe women's knowledge has been passed through the generations orally. Using ethnographic methods, data were gathered on traditional infant feeding practices from Ojibwe women (N = 44). Few of these traditions have been documented previously. Some traditions are similar to other indigenous cultures while others are culturally specific. Understanding traditional breastfeeding practices can provide valuable information for those working with indigenous people in a variety of settings, so that they create services that are consistent with traditional values. PMID- 12746032 TI - Incompatible expectations: the dilemma of breastfeeding mothers. AB - The maternal process of managing established breastfeeding and, ultimately, weaning the child from the breast was explored using the grounded theory method. Data were analyzed from interview transcripts from mothers, field notes, postal questionnaires from fathers, and individual and discussion group interviews with child health nurses within a Western Australian context. A common social problem emerged for women in the management of their breastfeeding when personal expectations were found to oppose the expectations of others. Being confronted with these incompatible expectations resulted in confusion, self-doubt, and guilt for mothers. These findings are discussed to assist health professionals support breastfeeding mothers. PMID- 12746034 TI - A global health opportunity with effective leadership. PMID- 12746036 TI - Tobacco counter-advertising: a review of the literature and a conceptual model for understanding effects. AB - The tobacco counter-advertising literature is reviewed as it relates to basic process questions concerning what makes counter-advertisements effective. Limitations in addressing (a) counter-advertisement content and the psychological mediators targeted, (b) counter-advertisement style and the affective reactions targeted, (c) prior smoking experience, and (d) other audience factors are enumerated. A theoretical model based on alcohol advertising research is presented to address those limitations. The model addresses the practical research question of predicting when tobacco counter-advertising will work by examining the independent influence of each of these enumerated factors, as well as how these factors operate in concert, qualifying each other. The model also addresses the process question of explaining how counter-advertising works by identifying affective and cognitive processes as mediators. By understanding the processes that underlie the qualified findings, one can better advise the designers of tobacco counter-advertisements how to be more effective. PMID- 12746037 TI - The effectiveness of gateway communications in anti-marijuana campaigns. AB - Successful anti-marijuana messages can be hypothesized to have two types of effects, namely persuasion effects, that is, a change in people's beliefs about using marijuana, and priming effects, that is, a strengthened correlation between beliefs and associated variables such as attitude and intention. This study examined different sets of anti-drug advertisements for persuasion and priming effects. The ads targeted the belief that marijuana is a gateway to stronger drugs, a belief that is often endorsed by campaign planning officials and health educators. A sample of 418 middle and high school students was randomly assigned to a control video or one of three series of ads, two of which included the gateway message in either an explicit or implicit way. Results did not support the use of the gateway belief in anti-marijuana interventions. Whereas no clear persuasion or priming effects were found for any of the ad sequences, there is some possibility that an explicit gateway argument may actually boomerang. In comparison to the control condition, adolescents in the explicit gateway condition tended to agree less with the gateway message and displayed weaker correlations between anti-marijuana beliefs and their attitude toward marijuana use. The results suggest that the gateway message should not be used in anti-drug interventions. PMID- 12746038 TI - Increasing intention to comply with pharmaceutical product instructions: an exploratory study investigating the roles of frame and plain language. PMID- 12746039 TI - The relationship of parental reinforcement of media messages to college students' alcohol-related behaviors. AB - Although alcohol consumption is a problem on the college campus, beliefs and behaviors predictive of alcohol use are in development in children as young as third grade. Because they develop partially in response to interpretations of media messages, for which parents can have an influence, this study examined how college students' (N=300) recollections of parental reinforcement of media messages associated with alcohol-related beliefs and behaviors. Structural equation modeling showed that recalled positive mediation negatively predicted skepticism, and positively predicted desirability and expectancies. Desirability of media messages predicted more positive norms perceptions, and a lack of skepticism predicted more positive expectancies. With age of first experimentation controlled, expectancies predicted heavier current drinking behavior. Norms did not predict behavior, and positive mediation did not predict norms. The study concludes that to the extent parental communication leads adolescents to interpret media messages less skeptically, they encourage adolescents to find alcohol portrayals appealing. This in turn appears to lead toward more risky behaviors. The results suggest that college-based anti-alcohol campaigns can benefit by acknowledging the appeal of competing messages and by including parents as a campaign target. PMID- 12746040 TI - A qualitative analysis of patient-centered dentistry in consultations with dental phobic patients. AB - Dental phobia is regarded as one of the greatest obstructions to adequate dental care. It has long been established that fearful dental patients are particularly sensitive to dentists' behavior and performance of dental care. There is a need for the establishment of a systematic theory of dentist-patient communication and new methods analyzing how dentists interact with their patients. In this qualitative study, thirty semi-structured interviews were conducted in 1998 and 1999 with five dentists (three male and two female). Dentists consulted on two occasions with 15 newly enrolled, consecutive dental phobic patients (2 male and 13 female) in a Swedish clinic specializing in the treatment of odontophobia. The time interval between consultation one and two was approximately 2-3 weeks. Analysis of the transcribed interviews was based by the principles of Grounded Theory. The study identified one core category, "Holistic perception and understanding of the patient", two categories, "The dentist's positive outlook on people" and "The dentist's positive view of patient contact", and six further subcategories. Findings support previous models of patient-centered medicine and contribute to a better understanding of how patient-centered dentists interact with dental phobic patients. PMID- 12746042 TI - Soluble ICAM-1, MCP-1, and MIP-2 protein secretion by rat pleural mesothelial cells following exposure to amosite asbestos. AB - Pleural inflammation is a sequela of exposure to toxic mineral fibers such as amosite asbestos. This inflammatory response involves the influx of leukocytes from the vasculature into the pleural space. Adhesion molecules such as intercellular adhesion molecule-1 (ICAM)-1 and chemokines such as monocyte chemoattractant protein-1 (MCP)-1 and macrophage inhibitory protein-2 (MIP)-2 are known to be important in pulmonary inflammation following inhalation of particulate matter. However, little is known about their role in pleural inflammation secondary to amosite asbestos exposure. Because the pleural mesothelial cell is believed to be a key target cell of asbestos exposure, the purpose of this study was to determine if ICAM-1, MCP-1, and MIP-2 proteins were secreted by these mesothelial cells following in vitro and in vivo exposure to amosite asbestos. Increased levels of ICAM-1 and MCP-1 protein were measured following 24 or 48 hours exposure of cultured rat pleural mesothelial cells to amosite fibers (1.5 to 5.0 micro g/cm(2)). Increased levels of ICAM-1, MCP-1, and MIP-2 protein were found in pleural lavage fluid from Fischer-344 rats exposed to amosite asbestos for 4 and 12 weeks and after a 12-week recovery period (following the 12-week exposure period). These findings suggest that the secretion of ICAM-1, MCP-1, and MIP-2 by rat pleural mesothelial cells may contribute to amosite-induced pleural inflammation. PMID- 12746043 TI - Different proliferative capacity of lung fibroblasts obtained from control subjects and patients with emphysema. AB - To characterize the possible role of a dysregulated proliferative capacity of pulmonary fibroblasts in insufficient tissue repair in lungs from patients with pulmonary emphysema, the authors undertook in vitro proliferative studies with pulmonary fibroblasts obtained from lung tissue of patients with emphysema. A comparison was made with fibroblasts from control subjects. The authors determined the in vitro proliferative capacity of fibroblasts at basal culture conditions and after modulation with interleukin-1beta, interferon-gamma, transforming growth factor-beta(1), and basic fibroblast growth factor. Proliferative capacity was determined by measurement of 5-bromo-2-deoxyuridine (BrdU) incorporation. BrdU incorporation by fibroblast cultures from both groups was very similar. Fibroblast cultures from control subjects, however, incorporated more BrdU after incubation with interleukin-1beta than cultures from patients with emphysema (P<.05). On the other hand, transforming growth factor beta(1) decreased incorporation of BrdU stronger in fibroblast cultures from control subjects than from patients with emphysema (P<.05). Thus, the proliferative capacity of fibroblast cultures isolated from lung tissue of patients with pulmonary emphysema is different from that of control subjects. Although the difference is small, it may be an essential contribution to the development of pulmonary emphysema that only occurs after repeated smoke-induced injury over many years of an individual's life. PMID- 12746044 TI - Effect of nitric oxide on lung surfactant secretion. AB - Lung surfactant is secreted from epithelial type II cells into alveolar airspace in response to airborne and circulating stimuli. Nitric oxide (NO) can be generated by constitutive and inducible nitric oxide synthases (cNOS and iNOS) in pulmonary endothelial and epithelial cells. The authors therefore examined the effects of NO on lung surfactant secretion using an isolated perfused rat lung model and primary culture of type II cells. Infusion of L-N(G)-nitroarginine methyl ester (L-NAME) (100 micro M), an inhibitor of cNOS and iNOS, via pulmonary circulation for 90 minutes resulted in a decrease of lung surfactant secretion (1.55%+/-0.15% in control versus 0.79%+/-0.16% in L-NAME-treated lungs, P <.05). However, aminoguanide, an inhibitor of iNOS, had no effect, indicating that the decline of lung surfactant secretion is due to the specific blockage of cNOS rather than iNOS activity in perfused lungs. A reduction of cGMP level by 1H [1,2,4] oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) (25 micro M), a specific inhibitor of guanylyl cyclase, inhibited surfactant secretion by 64%. Furthermore, KT5823 (1 micro M), an inhibitor of protein kinase G, depressed surfactant secretion by 40%. These results suggest that physiological concentrations of NO are required for lung surfactant secretion and NO-mediated secretion is at least partly via a rise of cGMP level and activation of protein kinase G. In primary culture of alveolar type II cells, spermine NONOate (SPER/NO), a NO donor, increased basal phosphatidylcholine (PC) secretion in a dose-dependent manner. Maximal stimulation was observed at 1 micro M. However, in the ATP-stimulated type II cells, SPER/NO displayed a biphasic effect on PC secretion. At low concentrations (0.1 to 1 micro M), SPER/NO increased ATP stimulated PC secretion, whereas at a high concentration (100 micro M), SPER/NO inhibited the secretion. The results suggest that NO may play an important role in lung surfactant secretion. PMID- 12746046 TI - Bronchial hysteresis: morphometric study on the rat lung. AB - The bronchus is an imperfect elastic structure because part of the energy transmitted in inspiration is not recovered in expiration. This phenomenon is known as bronchial hysteresis. The aim of this paper is to demonstrate bronchial hysteresis using morphometric techniques: differences in the wall and the bronchial lumen, between inspiration and expiration. Sixty disease-free Fischer rats were used, organized into 4 groups. The lungs from the first and second groups were fixed in inflation at 10 and 20 cm of H(2)O transpulmonary pressure (Ptp), respectively. Those taken from the third and fourth groups were fixed in deflation at 20 and 10 cm Ptp, respectively. The lungs were removed from the thorax, filled with air, and fixed via the pulmonary artery. A morphometric study of the following variables was performed: pulmonary volume (P(v)), dead space (DS) internal bronchial area (A(i)), which is the area of the bronchial lumen, and total wall area (WA(t)), which is the area of the bronchial wall. The lungs fixed in deflation gave higher values for all the variables than those fixed in inflation. The P(v), DS, and A(i) increased/decreased with the rise/fall of the Ptp; A(i) was the only variable to display statistically significant values (P<.05) when the Ptp was modified. WA(t) showed statistically significant differences between the lungs fixed in inflation and deflation and underwent no modification when the Ptp was varied. This study supports the hypothesis that bronchial hysteresis can be demonstrated using morphometric techniques because differences were observed in the lumen and wall of the bronchus between inflation and deflation. PMID- 12746045 TI - Increased spontaneous interleukin-10 release from alveolar macrophages in active pulmonary sarcoidosis. AB - The study was designed to determine whether alveolar macrophages (AM) in acute pulmonary sarcoidosis release in vitro the anti-inflammatory cytokine interleukin (IL)-10. To learn more about the coherence between IL-10 and proinflammatory cytokines in active sarcoidosis, the release of interferon (IFN)-gamma, macrophage inhibitory protein (MIP)-1alpha, and granulocyte-macrophage colony stimulating factor (GM-CSF) was studied and additionally compared to normal controls and patients with pneumonia and interstitial lung fibrosis. AM were obtained by bronchoalveolar lavage from 13 patients with active sarcoidosis, 8 patients with interstitial lung fibrosis, 10 patients with bacterial pneumonia, and 14 normal controls. The spontaneous and stimulated (tumor necrosis factor [TNF]-alpha, IL-1beta) cytokine release was measured in the supernatant of cultured AM by enzyme-linked immunosorbent assay (ELISA). Unstimulated AM from sarcoidosis patients released more IL-10, IFN-gamma, MIP-1alpha, and GM-CSF than normal controls and patients with pneumonia and interstitial lung disease. Stimulation with TNF-alpha or IL-1beta increased the MIP-1alpha and GM-CSF release from AM of normal controls and patients with pneumonia and interstitial lung disease: however, no further enhancement of MIP-1alpha and GM-CSF production was observed in AM from sarcoidosis patients. Exogenous IL-10 reduced the spontaneous and stimulated MIP-1alpha and GM-CSF release in sarcoidosis to a lesser extent than in controls and patients with fibrosis and pneumonia. The up regulated IL-10 in active pulmonary sarcoidosis may be a compensatory response to the enhanced expression of proinflammatory cytokines in order to down-regulate the inflammatory process. The results suggest an involvement of the anti inflammatory cytokine IL-10 in the immunopathogenesis of sarcoidosis. PMID- 12746047 TI - Maternally administered dexamethasone transiently increases apoptosis in lungs of fetal rats. AB - In late gestation, morphological maturation of fetal lung includes septal thinning of potential airspaces, a process accelerated by exogenous glucocorticoids. Apoptosis occurs in normal fetal lung. Glucocorticoids increase apoptosis in several tissues. The authors hypothesized that exogenous glucocorticoids would increase apoptosis in fetal lung, primarily in the interstitium. They administered dexamethasone (DEX), 1 mg/kg, or vehicle (Control) to pregnant rats at 19 days of gestation. Fetuses were delivered at 3, 7, 12, or 24 hours post injection. DEX decreased fetal body weight and lung weight, DNA, and protein 12 hours post injection. Using the terminal deoxynucleotide transferase-mediated dUTP nick-end labeling (TUNEL) reaction to label apoptotic cells in lung, they calculated an apoptotic index (AI, apoptotic cells/1000 total cells) for each fetus. Average DEX AI (3.6+/-2.6, mean+/-SD) was greater than Control (1.7+/-0.5) (P<.02). All DEX AIs were greater than Control AIs at 3, 7, and 12 hours, but were similar to Controls at 24 hours post injection. Apoptotic cells appeared to be interstitial, based on colocalization with vimentin staining. Presence of apoptotic cells was confirmed by electron microscopy and detection of the nucleosomal ladder pattern on DNA electrophoresis. The authors conclude that maternal administration of dexamethasone increases apoptosis in fetal lung, primarily in the interstitium. They speculate that apoptosis may contribute to morphological fetal lung maturation induced by endogenous glucocorticoids. PMID- 12746048 TI - Gene delivery to the lung by means of syngeneic fibroblasts: an experimental model. AB - An experimental model, in which exogenous gene expression in the lung is achieved, has been established. A fibroblast cell line was transfected with the lacZ gene and was administered to syngeneic mice by either intravenous or intratracheal injection. Enzyme-linked immunosorbent assay and histochemical detection of beta-galactosidase revealed efficient gene delivery to the lung by either route. Kinetic studies demonstrated that the expression peaked immediately after the injection, and this high level was maintained for 7 days by intratracheal and for 14 days by intravenous administration. This system may have potential relevance for certain experimental models requiring specific gene delivery to the lung. PMID- 12746049 TI - Is maternal copper supplementation during alveolarization protecting the developing rat lung against the adverse effects of maternal nicotine exposure? A morphometric study. AB - In a previous study, it was shown that maternal nicotine exposure during gestation and lactation interfered with alveolarization and resulted in gradual deterioration of the lung parenchyma, resulting in microscopic emphysema. The aim of this study was thus to investigate the long-term effects of maternal nicotine exposure (1 mg/kg body weight/day, subcutaneous [sc] from the onset of the phase of rapid alveolarization, which occur from postnatal day 4 in rats, on (1) the development of the gas-exchange area of the lungs of the offspring and, (2) whether maternal copper supplementation (1 mg/kg body weight/day, SC) during the same period of time will prevent the effect of maternal nicotine exposure on the development of the neonatal rat lung. Nicotine administration lasted until weaning on postnatal day 21. The day of birth was designated day 0. The offspring were exposed to nicotine via the mother's milk only. The experimental animals received no nicotine or copper after postnatal day 21. The lung tissue of the neonates was collected on postnatal days 14, 21, and 42 and prepared for morphometry. The results obtained show that maternal nicotine exposure had no influence on body weight, chest circumference, crown-rump length, and lung volume, but resulted in bigger alveolar volumes and suppressed alveolarization in the lungs of the offspring. Copper supplementation during this period of lung development reduced the adverse effect of maternal nicotine exposure on neonatal lung development. Even though copper reduced the adverse effects of maternal nicotine exposure during this phase of lung development, it did not prevent the induction of microscopic emphysema. PMID- 12746050 TI - Expression and activity of matrix metallo proteinases 2 and 9 and their inhibitors in rat lungs during the perinatal period and in diaphragmatic hernia. AB - During lung development, the extracellular matrix undergoes dynamic remodeling. Matrix metalloproteinases (MMPs), and tissue inhibitors of matrix metalloproteinases (TIMPs), are important enzymes that participate in regulating tissue remodeling. There is an abnormal balance of the synthesis and degradation of collagen and elastin in perinatal lung associated with congenital diaphragmatic hernia (CDH). This study was designed to (1) determine the expression and gelatinolytic activity patterns of MMPs 2 and 9 and TIMPs 1 and 2 in rat lungs during the perinatal period, and (2) to test the hypothesis that they are abnormal in nitrofen-induced CDH. Measurements were made using reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting, and zymography. The mRNA expression and activity of MMP 2 did not change significantly from embryonic day 16 to postnatal day 14. The most striking feature found was the rapid increase in the expression of MMP 9 soon after birth. Measurements were repeated on lung tissue isolated from embryonic rats with nitrofen-induced CDH. The expression and activity of MMPs and TIMPs were similar to control values and thus we conclude that these proteins appear not to be responsible for the altered extracellular matrix and morphological abnormalities noted in CDH lungs at birth. PMID- 12746052 TI - Physicochemical characterization of influenza viral vaccine loaded surfactant vesicles. AB - The goal of this study was to develop nonionic surfactant vesicles of influenza antigen for nasal mucosal delivery. The study describes the encapsulation of viral influenza vaccine antigen in nonionic surfactant vesicles using dehydration rehydration technique and investigation of the influence of the varying proportion of surfactant, cholesterol, and dicetyl phosphate on the morphology, particle size, entrapment efficiency, and in vitro antigen release from surfactant vesicles. The stability of the antigen was studied using SDS polyacrylamide gel electrophoresis and immunoblotting. The effect of cholesterol concentration and the method of lyophilization on antigen loading and in vitro release of antigen from surfactant vesicles also were studied. PMID- 12746051 TI - An attempt to modulate the microporous diffusion of a model polypeptide by altering its secondary structure. AB - The purpose of the present study was to determine whether intentional alteration of the secondary structure of a model polypeptide, conantokin-G, influenced the rate and extent of aqueous pore diffusion across a synthetic microporous membrane. Use of a microporous synthetic membrane allowed for analysis of polypeptide transport without the confounding variables of protein binding, acid- and/or enzyme-mediated degradation, endocytotic uptake, and enzymatic inactivation associated with a biological membrane. Conantokin-G was intentionally changed from its native random coil structure to the alpha-helix structure using calcium, and both structures were verified using circular dichroism. The alpha-helix structure of conantokin-G was retained even after additional free calcium was removed by equilibrium dialysis. Over the concentration range of 1.25 to 20 mM, there was a linear relationship between the solution calcium concentration and the percent of the alpha-helix conformer present. The apparent permeability, the apparent aqueous diffusion coefficient with and without inclusion of the Renkin function, and the hydrodynamic radii estimated by diffusion and a computer-software program were calculated for the random coil and alpha-helix structures of conantokin-G. Calcium-mediated conversion of conantokin-G to its alpha-helix structure did not significantly (p >.05) change its apparent permeability across a microporous membrane. It is suggested that perhaps complete conversion to the alpha-helix structure of only a fraction of the conantokin-G molecules (only 0.45 or 45% of the molecules can be converted to the alpha-helix structure at Ca(2+) concentrations >or= 20 mM) may have limited the extent of transport of the alpha-helix conformer. PMID- 12746053 TI - Correlation between gel structural properties and drug release pattern in scleroglucan matrices. AB - The drug delivery behavior of scleroglucan hydrogels was correlated with the structural features found by means of steady-state fluorescence studies. Theophylline at 0.4% in weight was used as the model drug and its release was measured using a Franz cell device. The results were fitted with a semi-empirical power law equation, finding significant differences between the 0.5% and higher than 0.8% scleroglucan concentrations (w/w). Four molecular fluorescent probes were used to test microenvironmental properties of the gel cavities, which will play a key role on the release of the pharmaceutical drugs. The results found by the fluorescence analysis are in good agreement with the ones obtained by release kinetics. PMID- 12746054 TI - Nonphospholipid vesicles of Carboplatin for lung targeting. AB - Carboplatin (CPt) was incorporated into nonphospholipid vesicles (NPLV) by the method of membrane dispersion and its content determined spectrophotometrically. The particle size was between 2.0-10.0 microm with a mean value of 3.26 microm and a span of 0.55; and the drug content was 3.965 mg/ml. Dynamic dialysis was used for in vitro release of CPt and the results indicated that the drug release t(1/2) was increased eight times, compared with the original drug. These parameters had little change after lyophilization. When tested with mice, the body distribution parameters showed obviously lung targeting characteristics. The S-180 lung neoplasm models were established by intravenous cancer cells in mice and the number of pulmonary nodules examined for evaluation of the therapeutic efficacy. Antitumor effect was increased and differed significantly (p <.05) compared with the original drug of the same dose. PMID- 12746056 TI - Formulation and in vivo evaluation of membrane-moderated transdermal therapeutic systems of nicardipine hydrochloride using carvone as a penetration enhancer. AB - A membrane-moderated transdermal therapeutic system (TTS) of nicardipine hydrochloride was developed using 2%w/w hydroxy propyl cellulose (HPC) gel as a reservoir system containing 8%w/w of carvone as a penetration enhancer. The permeability flux of nicardipine hydrochloride through ethylene vinyl acetate (EVA) copolymer membrane was found to increase with an increase in vinyl acetate content in the copolymer. The effect of various pressure-sensitive adhesives (MA 31, MA-38, or TACKWHITE A 4MED) on the permeability of nicardipine hydrochloride through EVA 2825 membrane (28%w/w vinyl acetate) or EVA 2825 membrane/skin composite also was studied. The results showed that nicardipine hydrochloride permeability through EVA 2825 membrane coated with TACKWHITE A 4MED/skin composite was higher than that coated with MA-31 or MA-38. Thus, a new TTS for nicardipine hydrochloride was formulated using EVA 2825 membrane coated with a pressure-sensitive adhesive TACKWHITE A 4MED and 2%w/w HPC gel as reservoir containing 8%w/w of carvone as a penetration enhancer. The bioavailability studies in healthy human volunteers indicated that the TTS of nicardipine hydrochloride, designed in the present study, provided steady-state plasma concentration of the drug with minimal fluctuations for 23 hr with improved bioavailability in comparison with the immediate-release capsule dosage form. PMID- 12746055 TI - Effect of liposomal delivery on in vitro antitumor activity of lipophilic conjugates of methotrexate with lipoamino acids. AB - Some selected lipophilic conjugates of the antifolate drug methotrexate (MTX) with lipoamino acids (LAA), previously described, were incorporated in liposomes with a different composition and charge (neutral, positive, or negative). The properties of the liposomal systems were determined. The inhibitory activity of the conjugates after incorporation in the vesicles was determined in a preliminary assessment against a human erythroleukemic cell line (K562 cells) and compared with the activity of the parent drug and of free conjugates. The influence of liposome surface charge and of the type of conjugate (i.e., in the carboxylic or ester form) on the biological effect is discussed. PMID- 12746057 TI - Studies on the development of colon targeted oral drug delivery systems for ornidazole in the treatment of amoebiasis. AB - The aim of the present study is to develop colon-targeted drug delivery systems for ornidazole using guar gum as a carrier. The core formulation containing ornidazole was directly compressed. Compression-coated tablets of ornidazole containing various proportions of guar gum in the coat were prepared. All the formulations were evaluated for hardness and drug content uniformity and were subjected to in vitro drug release studies. The amount of ornidazole released from tablets at different time intervals was estimated by the HPLC method. The compression-coated formulations released less than 8% of ornidazole in the physiological environment of stomach and small intestine. The compression-coated tablets with 85%, 75%, and 65% of guar gum coat released about 21%, 38%, and 73% of ornidazole, respectively, in simulated colonic fluids indicating the susceptibility of the guar gum formulations to the rat caecal contents. The results of the study show that compression-coated ornidazole tablets with either 65% (OLV-65) or 75% (OLV-75) of guar gum coat are most likely to provide targeting of ornidazole for local action in the colon owing to its minimal release of the drug in the first 5 hr. The ornidazole compression-coated tablets showed no change in physical appearance, drug content, or in dissolution pattern after storage at 40 degrees C/75% relative humidity for 6 months. PMID- 12746058 TI - Minoxidil-containing dosage forms: skin retention and after-rinsing hair-growth promotion. AB - Three kinds of topical dosage forms of minoxidil (MXD), namely vesicles, double emulsions, and an inclusion complex with hydoxypropyl-beta-cyclodextrin (HP-beta CD), were prepared. The skin retention of MXD in the preparations was evaluated in vitro using hairless mouse skins. After applying the preparations onto the skin and rinsing it, the amount of the drug left on the skin was determined using HPLC. Retention was the highest when the drug was encapsulated in cationic vesicles. Nonionic vehicle, the double emulsion, and HP-beta-CD left no significant amount of the drug after rinsing the skin. Thus, an ionic interaction between the cationic vehicle and negatively charged skin is likely responsible for the relatively high skin retention. In vivo hair growth-promotion effect of each dosage form was investigated, in which the sample application onto the clipped backs of female mice (C57BL6) and the subsequent rinsing of the backs were done once a day for 30 days. Only MXD in the cationic vesicles had hair growth promotion effect, possibly due to significant skin retention. PMID- 12746062 TI - Material safety data sheets: their use in Quebec workers' compensation appeals involving neurotoxic chemical exposures. PMID- 12746063 TI - National follow-up of occupational health services system in Finland. PMID- 12746065 TI - Characterization of agricultural tasks performed by youth in the Keokuk County Rural Health Study. AB - Injury and illness among youth working on farms are important problems. The types of farm tasks performed by children and the ages at which they begin these activities have not been well characterized. This cross-sectional study characterized agricultural work performed by adolescents in a rural Iowa county to better understand the patterns and extent of exposures to agricultural risk factors. This information will help to develop prevention strategies for agriculture-related injury and illnesses for children working on farms. The Keokuk County Rural Health Study is a prospective cohort study of randomly selected households in Iowa. In Round 2, all youth, aged 12 to 17 years participating in this population-based study, were asked about their use of farm machinery, work with livestock, pesticide handling, and other farm activities. The age at which they actually began these activities, the age they believed youth should start these activities, and sources of health and safety training they received were also ascertained. Adults in the study were asked the same questions about youth. Matched parental reports and opinions were compared to their children's reports and opinions using McNemar's chi-square tests. A total of 143 youth and 684 adults with farming experience completed the interviews. There were 118 pairs of parents and youth. Fifty percent of male youth, and 18 percent of females had performed agricultural work at some time in their life. Twenty-five percent of all male youth, and 5 percent of females were currently working on farms. Close to 30 percent had driven tractors, all-terrain vehicles, and pick-up trucks. Despite the legal prohibition of hazardous work by children under age 16, several younger children reported that they had driven a self propelled combine, worked in silos, or handled and applied fertilizers in the past 12 months. Youth began riding on tractors at a mean age of 7. The mean ages for driving tractors and all-terrain vehicles were 11 and 10, respectively. The mean age for driving a self-propelled combine was 13. Children began applying or handling fertilizers at the age of 12. There were discrepancies between parent and youth reports regarding the frequencies, starting age, and opinions relative to performing agricultural tasks. These results suggest the need for implementing guidelines, particularly for age appropriate agricultural tasks. PMID- 12746067 TI - Asbestos release during removal of resilient floor covering materials by recommended work practices of the resilient floor covering institute. AB - The release of asbestos during maintenance and removal of resilient floor covering is of concern to health professionals and many regulators. This study assesses the asbestos levels observed during removal of resilient floor covering products using the "Recommended Work Practices" (1995) of the Resilient Floor Covering Institute or other methods requiring containment (Controls). The 1995 "work practices" require wet removal or dry heat removal but do not require the use of respirators. Wet removals of sheet vinyl/separated backing, 12" x 12" vinyl asbestos tile/mastic, and 9" x 9" asphalt tiles/mastic were conducted and the air was sampled during each procedure. Settled dust samples were collected at the sites of RFCI square tile removal and pieces of each type of tile were broken in a mini-enclosure to evaluate asbestos emissions. Analyses of the air samples collected during the removals showed that the RFCI methods did not produce asbestos counts significantly different from the Control methods requiring containment. Only a small number (0.7%) of fibers and structures, counted and measured by Analytical Transmission Electron Microscopy, would have been counted using the rules for Phase Contrast Microscopy in the 7400 method specified by Occupational Safety and Health Administration regulations. This indicates workers in similar situations without respirators are likely to have unknown exposure levels. A high percentage of these fibers and structures are 5 micrometers or less in length, smaller than 0.5 micrometer in diameter, and are easily inhaled. The RFCI air sample and settled dust data may cause regulators to consider requiring respiratory protection, cleanup procedures, and methods to control asbestos migration. Other areas that might be addressed are clearance levels and their measurement, removal area size, bulk sample analysis by transmission electron microscopy if polarized light microscopy reports less than 1 percent asbestos, better worker exposure evaluation, and supervisor/worker training in accordance with the Model Accreditation Plan. PMID- 12746066 TI - Workplace airborne hexavalent chromium concentrations for the Painesville, Ohio, chromate production plant (1943-1971). AB - Hexavalent chromium [Cr(VI)] is recognized as an inhalation carcinogen, based primarily on the increased incidence of lung cancer among occupationally exposed workers. To assess the carcinogenic potency of Cr(VI), both the U.S. Environmental Protection Agency and the Occupational Safety and Health Administration have relied on data from a 1930s cohort of workers from the Painesville, Ohio, chromate production plant. However, the exposure information for this cohort has several shortcomings. In an effort to provide better exposure information, we present here recently identified historical exposure data for the Painesville workers. More than 800 measurements of airborne Cr(VI) from 23 newly identified surveys conducted from 1943 to 1971 are presented. The results indicate that the highest Cr(VI) concentrations recorded at the plant occurred in shipping (e.g., bagging of dichromate), lime and ash, and filtering operations, with maximum yearly average Cr(VI) concentrations of 8.9, 2.7, and 2.3 mg/m(3), respectively. The locker rooms, laboratory, maintenance shop, and outdoor raw liquor storage areas had the lowest average Cr(VI) air concentrations over time, with yearly average concentrations that rarely exceeded the historical and current Threshold Limit Value TLV(R) of 0.05 mgCr(VI)/m(3) (0.1 mgCrO(3)/m(3)). Concentrations generally decreased in the plant over time. The average airborne concentration of Cr(VI) in the indoor operating areas of the plant in the 1940s was 0.72 mg/m(3), that from 1957 through 1964 was 0.27 mg/m(3), and that from 1965 through 1972 was 0.039 mg/m(3). Although in some ways limited, these data are of sufficient quality to allow for exposure reconstruction for workers employed at this plant from 1940 to 1972, and to provide the basis for an improved cancer risk assessment. PMID- 12746068 TI - Video exposure assessments demonstrate excessive laboratory formaldehyde exposures. AB - Video exposure assessments were conducted in a comparative anatomy laboratory using formaldehyde-preserved sharks and cats. Work in the facility using time integrated samplers indicated personal and area concentrations generally below the current OSHA permissible exposure limit. However, complaints about room air quality were frequent and routine. Using a photoionization detector with an integral data logger, total ionizables present were sampled as a surrogate for formaldehyde. After synchronizing time tracks from the datalogger concentrations with simultaneously created videotapes of laboratory tasks, composite video exposure overlays were generated. Use of this video exposure method revealed very short-lived, excessively high peak exposure events, whereas conventional time weighted averages indicated the majority (30/32) of personal exposures were below the OSHA limit of 0.75 ppm. These legally acceptable exposure levels were associated with self-reported symptoms of burning nose and eyes and eye irritation. Thus, transient peak formaldehyde concentrations not detected by longer term averaging studies could be responsible for the health effects reported. The video exposure monitoring method demonstrated that close dissection work, opening peritoneal cavities, and specimen selection activities were most likely the causes of elevated student exposures. Teaching assistants' exposures were the highest, exceeding OSHA limits on several occasions. The utility of the video monitoring method for conducting enhanced, critical task exposure assessments is discussed. PMID- 12746069 TI - Analysis of sampling line bias on respirable mass measurement. AB - This study investigated the bias introduced by an inlet sampling line on a respirable mass monitor. The 1.5-m electrically conductive, flexible sampling line conducts aerosol at a flow rate of 2.2 Lpm from a helmet-mounted inlet to a waist-mounted sensor for mass concentration measurement. Particulate transport was modeled for each section of the sampling line and considered the effects of diffusion, gravitational settling, and inertial impaction. An estimate of respirable mass concentration measured with the sampling line was determined by integrating assumed workplace aerosols with the transport curves. The bias introduced by the sampling line was then calculated by dividing the difference between the respirable mass concentration with and without the sampling line by that without the sampling line. For the current sampling line, in which the inner diameter is 4.83 mm, bias was calculated as -0.3 percent, -2.4 percent, -4.6 percent, and -6.7 percent for four test aerosols with mass median aerodynamic diameters of 0.6 microm, 4 microm, 12 microm, and 30 microm, respectively. Optimization studies suggest that increasing the sampling line with a larger inner diameter by a factor of 1.25 to 1.75 will minimize bias to below -3.0 percent. An experimental study confirmed that bias due to the presence of the sampling line is small. PMID- 12746070 TI - Work-related injuries in residential and drywall carpentry. AB - Findings are reported on the first two years of an active injury surveillance project designed to test the utility of active injury investigations in identifying causes of injury among a large cohort of carpenters who did residential building and drywall installation. Occupational Safety and Health Administration recordable injuries were reported by participating contractors. Injured union carpenters were interviewed by experienced journeymen trained in a standard questionnaire protocol. Enumeration of workers and hours worked were provided by the union. These data allowed the definition of a dynamic cohort of 4429 carpenters, their hours worked, detailed information on the circumstances surrounding recordable injuries, and possible preventive measures from the perspectives of the injured worker and an experienced journeyman investigator. The overall estimated injury rate (16.9 per 200,000 hours worked) was considerably higher than recent Bureau of Labor Statistics rates despite less than complete ascertainment of injuries. Injuries most commonly involved being struck by or against something, manual materials handling injuries, and falls. Manual materials handling injuries often involved very heavy objects or tasks and were injuries carpenters most often reported needs for adequate help and coordinated team work to prevent. Falls from heights occurred from a variety of surfaces and were not just injuries of inexperience. Carpenters reported the need for more attention to common fall protection practices, such as the use of more toe boards and guardrails. Poor housekeeping was involved in the majority of same level falls, as well as some manual materials handling injuries. PMID- 12746072 TI - Health hazards to park rangers from excessive heat at Grand Canyon National Park. PMID- 12746073 TI - An endotoxin exposure in the food industry. PMID- 12746074 TI - Data collection and "real-time" learning using handheld computers. PMID- 12746075 TI - Stretching at work for injury prevention: issues, evidence, and recommendations. PMID- 12746076 TI - Safety in design: a proactive approach to construction worker safety and health. PMID- 12746078 TI - A comparison of the sampling efficiencies of a range of atmosphere samplers when collecting polymeric diphenylmethane di-isocyanate (MDI) aerosols. AB - Polymeric diphenylmethane di-isocyanate (PMDI) is a precursor or an intermediate in the manufacture of pure MDI and is used in many industrial applications. Potential health effects of diisocyanates are generally considered to include irritation and respiratory sensitization, dictating the need for low occupational hygiene standards and robust hygiene monitoring methods. A wide range of methods has been developed or adopted for this purpose but questions concerning their ability to accurately sample MDI aerosols in the occupational environment have often been raised. In order to provide such information, studies have been conducted to compare several MDI sampling systems: the Institute of Occupational Medicine (IOM) 25-mm open-faced inhalable dust sampler; the 13-mm Millipore Swinnex Cassette sampler; the 37-mm open-face Millipore cassette; the midget impinger, the glass tube containing glass wool, and two direct reading paper tape monitors. The program was comprised of two phases, the first being a preliminary comparison of the collection efficiencies of the IOM, 13 mm and midget-impinger at a range of orientations to air flow, aerosol particle sizes, and sampling flow rates, using inert polyethylene glycol aerosols. The second phase compared all samplers operating according to each manufacturer's recommendations and sampling PMDI aerosols at a range of particle sizes and concentrations. All studies were conducted in a wind tunnel. All filter methods performed well in atmospheres containing small particles except impingers that required a filter backup. In general, the variability of all the samplers was high for larger particle size ranges. Direct reading monitors showed low efficiencies. PMID- 12746079 TI - Air sampling methodology for asphalt fume in asphalt production and asphalt roofing manufacturing facilities: total particulate sampler versus inhalable particulate sampler. AB - In 2000, the American Conference of Governmental Industrial Hygienists (ACGIH(R)) changed its 1971 threshold limit value (TLV) for 8-hour time-weighted average (TWA) exposure to asphalt from 5 mg/m(3) total particulate (generally < or =40 micrometer [microm] diameter) to 0.5 mg/m(3) inhalable particulate (< or =100 microm aerodynamic diameter) as benzene-soluble aerosol. To date, no inhalable particulate sampling method has been standardized and validated for asphalt fume. Furthermore, much of the historical data were collected using total particulate samplers, and the comparability of total versus inhalable size fractions of asphalt fume is not known. Therefore, the present study compared results from two types of asphalt fume samplers: 1) a traditional total particulate sampler with a 37-mm filter in a closed-face cassette with a 4-mm orifice (NIOSH 5042) versus (2) an inhalable particulate sampler designed by the IOM with a 15-mm orifice. A total of 75 simultaneous pairs of samples were collected, including personal and area samples from 19 roofing and asphalt production facilities operated by 7 different manufacturers. Each sample was analyzed for total mass collected and for benzene-soluble mass. Data from the two sampling methods (total versus inhalable) were comparable for asphalt fumes up to an aerosol concentration of 10 mg/m(3). However, we conclude that the traditional total particulate method is preferable, for this reason: The vast majority of asphalt fume particles are <12.5 microm in diameter. The traditional sampler is designed to collect primarily particles < or =40 microm, while the IOM sampler is optimized for collecting particles < or =100 microm. Thus, the traditional sampler is less likely than the IOM sampler to collect the larger-size fraction of airborne particles, most of which are non-asphalt dust. PMID- 12746080 TI - The application of diffusive sampling combined with thermal desorption in occupational exposure monitoring--field evaluation. AB - Diffusive sampling combined with thermal desorption (DSTD) is considered useful in monitoring workers' exposures to organic vapors, not only due to its simplicity of operation in the field but, also, its simplifying the sample treatment, eliminating the use of extracting solvent, and increasing analytical sensitivity. Herein, the feasibility of applying DSTD in field conditions was assessed through a comparison of the results with those from the well-accepted active/charcoal method. Side-by-side active and diffusive (tube-type, Tenax TA as the adsorbent) samples, both personal and area, were collected and analyzed for xylenes, ethyl acetate, styrene, and n-butyl acetate in four different industrial settings. Statistically significant correlations were found between the two methods in personal samples with the correlation coefficients of 0.92, 0.90, 0.83, 0.88, and 0.97 for m&p-xylenes, o-xylene, n-butyl acetate, styrene, and ethyl acetate, respectively. A paired t-test revealed significant difference between the two methods for n-butyl acetate and styrene. For area sampling, statistical differences (p < 0.05) were found between the two methods, except for xylenes. The DSTD method had, mostly, a lower concentration than the active method, with the bias ranging from 10.2 to -54.4 percent. The possible causes for the discrepancies are discussed. These results suggest that though DSTD protocols can be considered as a simple approach for screening workers' exposures to volatile organics, they should be applied with caution since diffusive sampling is potentially affected by various environmental conditions and adsorptive characteristics. PMID- 12746081 TI - Nail gun injuries among construction workers. AB - Pneumatic nail guns greatly increase worker productivity and are extensively used in wood frame building construction, with especially high use in residential construction. One surveillance report of nail gun injuries in Washington State has been published; however, other literature consists largely of case reports and case series in trauma journals. The major objective of the current study was to investigate the occurrence of nail gun-associated injuries among construction workers and to identify preventable work-related factors associated with these injuries. Nail gun-related injuries occurring among a cohort of 13,347 carpenters in Ohio who worked union hours during the time period January 1, 1994, until September 30, 1997, were identified by matching the cohort with workers' compensation claims made to the Ohio Bureau of Workers' Compensation. We also analyzed workers' compensation claims for North Carolina Home Builders Association members for the period July 1996-November 1999 to identify nail gun related injuries. Analyses included stratified analyses of claims by nature and body part injured, calculation of nail gun injury rates, and analyses of free text descriptions of injuries. Overall, nail gun injuries were responsible for 3.9 percent of workers' compensation claims with 8.3 percent to 25.5 percent of claims involving paid lost work time. The overall rate of nail gun injuries (cases per 200,000 work hours) was 0.33 in North Carolina and 0.26 in Ohio, reflecting the greater concentration of wood frame construction workers in the North Carolina population studied. Higher rates of injury were observed for carpenters in North Carolina and among residential carpenters in Ohio. The predominant body part injured was the hands/fingers, with 80 to 89 percent of injuries being nail punctures. Analyses of free text information for puncture injuries found approximately 70 percent of injuries to occur during the framing/sheathing stage of construction. Our data suggest that approximately 69 percent of puncture injuries may be due to an inadvertent gun discharge or misfire, preventable in large part by the use of sequential triggers. Worker training and education also are important components of nail gun injury prevention. PMID- 12746082 TI - Comparative occupational exposures to formaldehyde released from inhaled wood product dusts versus that in vapor form. AB - Particle boards and other wood boards are usually made with formaldehyde-based resins. Woodworkers are thus exposed to formaldehyde in vapor form as well as from airborne dust once it enters their respiratory tract. These workers remain exposed to formaldehyde released from the dust still present in their upper respiratory tract, even after their work shift. In assessing the risk associated with formaldehyde exposure, one needs to consider the relative importance of these two sources of exposure. This study proposes two kinetic models to estimate and compare the exposures. For various exposure scenarios, one model predicts the amount of formaldehyde absorbed from the ambient vapor form and the other predicts the amount absorbed by the respiratory tract upon its release from wood product dust. Model parameters are determined using data from published studies. Based on a daily work shift of 8 hr, with a dust concentration in air of 5 mg/m(3) and a formaldehyde concentration bound to dust of 9 microg/mg, model simulations predict that the amount of absorbed formaldehyde released from wood dust is approximately 1/100 of the amount absorbed from the ambient vapor form at a concentration level of 0.38 mg/m(3) (0.3 ppm). Since the formaldehyde concentration in wood dust used above is much higher than usually observed while the dust and vapor form formaldehyde concentrations are of the order of acceptable upper values, these results indicate that the formaldehyde exposure from wood dust is comparatively negligible. PMID- 12746083 TI - Prototype sampling system for measuring workplace protection factors for gases and vapors. AB - A prototype sampling system for measuring respirator workplace protection factors (WPFs) was developed. Methods for measuring the concentration of contaminants inside respirators have previously been described; however, these studies have typically involved continuous sampling of aerosols. Our work focuses on developing an intermittent sampling system designed to measure the concentration of gases and vapors during inspiration. This approach addresses two potential problems associated with continuous sampling: biased results due to lower contaminant concentrations and high humidity in exhaled air. The system consists of a pressure transducer circuit designed to activate a pair of personal sampling pumps during inspiration based on differential pressure inside the respirator. One pump draws air from inside the respirator while the second samples the ambient air. Solid granular adsorbent tubes are used to trap the contaminants, making the approach applicable to a large number of gases and vapors. Laboratory testing was performed using a respirator mounted on a headform connected to a breathing machine producing a sinusoidal flow pattern with an average flow rate of 20 L/min and a period of 3 seconds. The sampling system was adjusted to activate the pumps when the pressure inside the respirator was less than -0.1 inch H(2)O. Quantitative fit-tests using human subjects were conducted to evaluate the effect of the sampling system on respirator performance. A total of 299 fit-tests were completed for two different types of respirators (half- and full-facepiece) from two different manufacturers (MSA and North). Statistical tests showed no significant differences between mean fit factors for respirators equipped with the sampling system versus unmodified respirators. Field testing of the prototype sampling system was performed in livestock production facilities and estimates of WPFs for ammonia were obtained. Results demonstrate the feasibility of this approach and will be used in developing improved instrumentation for measuring WPFs. PMID- 12746085 TI - Should opioid analgesics be used in the management of chronic pain in opiate addicts? PMID- 12746086 TI - State substance abuse treatment gaps. AB - This study estimated the adequacy of state substance abuse treatment rates relative to treatment needs. The investigators created composite drug and alcohol treatment need indexes from explicit-mention mortality and substance-defined arrest rates. The indexes were reliable and had evidence of construct validity, but alternative population-at-risk and survey-based need measures did not fair as well. States varied substantially in per capita alcohol and drug treatment needs, although the two did not correlate with each other. While the need indexes correlated significantly with state treatment rates, the adequacy of state treatment rates varied greatly. States with the largest treatment gaps were in the South, Southwest, and northern plains and mountain regions. The failure of the Block Grant formula to reflect the needs of rural states with high-risk minority populations may contribute to disparities in access to services. PMID- 12746087 TI - Comparison of desipramine or carbamazepine to placebo for crack cocaine-dependent patients. AB - The authors compared the effects of desipramine or carbamazepine to placebo in an intensive outpatient program for cocaine abuse. Subjects recruited from an urban drug treatment program were randomly assigned to a double-blind, placebo controlled, eight-week trial of desipramine, carbamazepine, or placebo. Patient ratings, urine drug screens, and blood samples were obtained weekly. Using survival analysis, the three groups did not differ in time to drop out of treatment. While subjects improved over time on all self-ratings related to cocaine use, mood, and craving, only two items related to mood were significantly different over time as a function of treatment group. Subjects in the two treated groups reported significantly more improvement on self-ratings of depression and irritability. No treatment differences were noted for sustained abstinence or for proportion of positive urine drug screens. Desipramine subjects who attained a minimum blood level were retained in treatment significantly longer than placebo or other non-compliant treatment groups. This finding supports previous reports of a possible role for desipramine in cocaine abuse treatment. PMID- 12746088 TI - Compliance to supervised disulfiram therapy: a comparison of voluntary and court ordered patients. AB - We hypothesized that court mandate would significantly enhance compliance with supervised disulfiram therapy. We conducted a twelve-week prospective study of outpatient compliance with court-ordered, monitored disulfiram treatment as compared to voluntary, monitored treatment. The court ordered group (n=19) was significantly more compliant than the voluntary group (n=22). Legally mandated subjects attended an average of 87% (+/-21%) of scheduled visits, versus 42% (+/ 35%) for the group without court order. Court mandate roughly doubles the compliance rate of monitored disulfiram therapy, effectively enhancing clinic attendance during the first twelve weeks of treatment. PMID- 12746089 TI - Psychiatric disorders and cigarette smoking among child and adolescent psychiatry inpatients. AB - We surveyed 120 patients admitted to a child and adolescent psychiatric inpatient unit regarding their cigarette smoking history, reasons for smoking, patterns of use, and psychiatric comorbidity (chart review). We administered the Social Phobia and Anxiety Inventory and the modified Fagerstrom Tolerance Questionnaire. Only six of 47 current smokers had a chart diagnosis of nicotine dependence. Logistic regression derived that the odds ratio for being a current smoker was: Caucasian race=5.56,older age=1.3, depressive disorders=4.02, conduct disorder (CD)=12.96, cannabis abuse/dependence=24.6. Hence, cigarette smoking is highly prevalent but underdiagnosed in this population. PMID- 12746090 TI - Reducing hospitalizations and arrests for substance abusers. AB - This four-year outcome study examines the results of the VA Contracted Residential Substance Abuse Treatment for veterans with substance abuse problems. Race, marital status, housing status, and exposure to combat in the military characterized the male veteran subjects. Hospitalization rates and arrest rates were tracked for two years prior to and two years post-residential treatment. The authors also examined treatment outcomes for those who completed outpatient substance abuse treatment after an episode of residential care. This study shows a 30% reduction of hospitalizations and arrests following residential treatment. Those veterans who completed both residential and outpatient treatment had the best outcomes in terms of number of hospitalizations and arrests in a two-year follow-up. PMID- 12746092 TI - State policy and practice regarding substance abuse treatment services for medicaid recipients with disabilities. AB - This article presents the results of an exploratory study conducted to identify best practice Medicaid managed care models for people with disabilities who need substance abuse treatment services. These results suggest that there is wide variation in the managed care strategies that states use to provide substance abuse treatment services to the SSI disabled population, that state policymakers are often focused on general program management issues rather than addressing specific issues related to providing substance abuse treatment services to people with disabilities, and that although managed care theoretically offers opportunities for creativity,this practice does not appear to be widespread under current Medicaid managed care arrangements for people with disabilities. PMID- 12746091 TI - Factor Analysis of the Attention Deficit Scales for Adults (ADSA) with a clinical sample of outpatient substance abusers. AB - Individuals with adult symptoms of ADHD who are in treatment for substance abuse have been identified as being at greater risk of failure to complete treatment and relapse. The identification of such clients must thus become an important part of assessment and treatment planning. The Attention Deficit Scale for Adults (ADSA) is one device designed to assess ADHD symptomology in adults. This study investigated the application of the ADSA in a substance-abusing population by assessing its psychometric properties with a clinical sample of 268 adult patients (170 males, 92 females, gender missing for 6) in an outpatient substance abuse treatment program. The findings indicate that the ADSA is unidimensional with this population yet possess high reliability (alpha=.93 for entire sample:.89 for males,.94 for females) and correlate consistently with DSM-IV diagnostic criteria. The data tentatively support the use of the ADSA with substance-abusing populations as a reliable and valid assessment tool for clinical use and research. PMID- 12746094 TI - Genetic counseling in assisted reproduction: a case of cystic fibrosis identified after two successful intracytoplasmic sperm-injection pregnancies. PMID- 12746095 TI - Intracytoplasmic sperm injection: stiletto conception or a stab in the dark. AB - To describe the importance of molecular and cellular analyses in intracytoplasmic sperm injection (ICSI) the authors review the literature on biological challenges in ICSI and associated techniques. Several matters can be proposed in molecular and cellular challenges in ICSI for safety and efficacy: (1) a reliable and convenient animal model for understanding the molecular and cellular basis of human ICSI must be established, and molecular and cellular analysis of the first cell cycle of human fertilization should be better understood; (2) a proper assay for human sperm function that contributes to the indication for ICSI should be developed; and (3) de novo and transmitted genetic security in ICSI should be examined. PMID- 12746096 TI - Activation of endothelial nitric oxide synthase in contralateral testis during unilateral testicular torsion in rats. AB - There are controversies about the injury of the contralateral testis during unilateral testicular torsion (UTT). An autonomic reflex arc between bilateral testes has been proposed. The authors focused on the involvement of nitric oxide (NO) in the contralateral testis during UTT. Eight-week-old male Wistar rats underwent unilateral torsion (1 h)-detorsion (up to 24 h). NO synthase (NOS) activity was detected as NADPH-diaphorase activity after fixation by paraformaldehyde. N-nitro-L-Arginine methyl ester (L-NAME, 20 mg/kg) was injected intravenously to the other group of rats. To evaluate the testicular injury, proteolysis of alpha-fodrin production was detected by Western blotting. Apoptosis of the germ cells was evaluated by TUNEL. Long-term effect on spermatogenesis was evaluated by flow cytometry at 60 days after UTT. Transient activation of NOS was detected following the proteolysis of alpha-fodrin in the contralateral testis. L-NAME inhibited these alterations. NADPH-diaphorase activity and eNOS immunoreactivity were co-localized in the endothelial cells. These reactions were not observed in other organs. There was neither enhanced apoptosis nor deteriorated spermatogenesis in the contralateral testis during and 60 days after UTT. In the contralateral testis, eNOS-derived NO regulates the vasomotor function against unilateral testicular torsion, whereas it acts slightly cytotoxic. These results suggest the possible involvement of a testis specific neurovasomotor reflex between the bilateral testes. PMID- 12746098 TI - Complication rate of microsurgical varicocele ligation without delivery of the testis. AB - It is commonly assumed that the complication rate of microsurgical varicocele repair is lower than that of nonmicrosurgical techniques, particularly since magnification allows for preservation of lymphatic vessels and, thus, a theoretically lower incidence of hydrocele. However, review of the literature reveals relatively few papers on this subject. The authors reviewed their experience to determine the incidence of complications following microsurgical varicocele repair without delivery of the testicle in the adult population. From 1 July 1997 to 1 September 2001 139 men underwent microsurgical varicocele ligation without delivery of the testicle, with a mean follow-up of 22 months. Of the 139 men, 4 (2.9%) had complications. One (0.7%) had a recurrence, 1 (0.7%) had a wound infection, 1 (0.7%) had epididymitis, and 1 (0.7%) had an unintentional injury of the testicular artery. None of the men developed a hydrocele. The published complication rate for nonmicrosurgical varicocle ligation ranges from 5.4 to 7.2%, with the most common complication being hydrocele formation. In the authors' series, the overall complication rate was 2.9%, with no patient developing a hydrocele. Thus, microsurgical varicocele ligation without delivery of the testicle results in a lower complication rate and a lower incidence of hydrocele than nonmicrosurgical techniques. PMID- 12746097 TI - Growth hormone, insulin growth factor-1, and igf binding protein-3 axis relationship with bone mineral density among healthy men. AB - The aim of this study was to investigate serum levels of growth hormone (GH), insulin growth factor-I (IGF-I), and insulin growth factor binding protein-3 (IGFBP-3) in 363 healthy caucasian men with and without decreased bone density, who had never experienced fractures. Mean age was 51+/-8.7 years. Height and weight were measured and BMI was calculated using the formula weight (kg)/height (m(2)). Bone mineral density (BMD) was assessed: in 4 skeletal sites (lumbar spine [LS], femoral neck [FN], Ward's triangle [WT], and trochanter [T]) using dual-energy X-ray absorpsiometry (DEXA). After an overnight fasting, blood samples were taken at 8:00 a.m. Serum concentrations of GH, IGF-I, and IGFBP-3 were measured using the immunofunctional (GH) and IRMA (IGF-I and IGFBP-3) methods. The BMD at the 4 skeletal sites is expressed as mean value+/-SD in g/cm(2) and T score. Forty-four men (11%) had bone mineral density (BMD)<-2.5 SD (T score). Mean GH, IGF-I, and IGFBP-3 levels were 0.2+/-0.1, 186.1+/-177.3, and 4990+/-1460 ng/mL, respectively. There were no significant differences between men with normal BMD and men with reduced BMD concerning GH, IGF-I, and IGFBP-3 measurements. In normal men (319), mean GH, IGF-I, and IGFBP-3 levels were 0.4+/ 0.1, 192+/-87, and 4960+/-1530 ng/mL, respectively. In the subgroup with reduced BMD (44), mean GH, IGF-I and IGFBP-3 levels were 0.2+/-0.1, 179+/-72 and 5230+/ 1270 ng/mL, respectively. An age-dependent attenuation of GH, IGF-I, and IGFBP-3 levels was also found. No correlation was revealed between BMD and GH in the 4 skeletal sites tested. On the contrary, a positive correlation was established between BMD and IGF-I levels in 3 skeletal sites (LS, FN, T). The same was true between BMD and IGFBP-3 in 2 skeletal sites (LS, FN). In conclusion, 11% of Greek healthy males had decreased bone density. No fractures were demonstrated in any individuals. No significant differences were found between men with normal and reduced BMD, with regards to serum GH, IGF-I, and IGFBP-3, although these levels decreased with age. No correlation was found between BMD and GH levels in the 4 skeletal sites. A positive correlation was found between BMD and IGF-I levels in 3 skeletal sites and IGFBP-3 in 2 skeletal sites. PMID- 12746099 TI - Extracorporeal shockwave therapy for peyronie disease. AB - While surgery is the mainstay of therapy for Peyronie disease requiring correction of angulation, interest has grown in the application of extracorporeal shockwave therapy (ESWT) as a minimally invasive approach. This article reviews the current literature reporting the use of ESWT for Peyronie disease. PMID- 12746100 TI - Successful treatment of retrograde ejaculation with amezinium. AB - The effect of amezinium, a new type of antihypotensive agent, on retrograde ejaculation was evaluated in 3 patients with retrograde ejaculation. The patients received 10 mg amezinium orally once a day. All patients achieved antegrade ejaculation. Semen analyses revealed 6-50 x 10(6)/mL (mean 28.7 x 10(6)/mL) sperm with a motility of 20-50% (mean 36.7%). The wives of 2 patients became pregnant within 6 months of the initial treatment. None of the patients had any side effects. It would appear that amezinium is a useful treatment for retrograde ejaculation. PMID- 12746101 TI - Improvement of sexual activity, pregnancy rate, and low plasma testosterone after bilateral varicocelectomy in impotence and male infertility patients. AB - To evaluate the effects of bilateral varicocelectomy on sexual activity, testicular volumes, semen quality, and serum hormone levels in impotence and male infertility patients, 48 patients were studied from an outpatient clinic from May 1998 to March 2001. The mean age was 37+/-5.9; 16 patients were complaining of erectile dysfunction and 32 patients were complaining of male infertility. The mean duration of impotence was 3.3+/-2.4 years and for male infertility was 3.8+/ 3.2 years. Sexual and reproductive history was taken for erectile dysfunction and male infertility patients. General, local examination, and laboratory investigations were done for all patients. Preoperative and postoperative testicular volumes; semen parameters, including semen volume, sperm count, and motility; and morphology and hormonal parameters, including LH and FSH, and testosterone levels were measured. All patients were followed up for 3-36 months after varicocele repair. Left and right testicular volume was improved in impotence and male infertility patients and fertility groups, but this improvement was not statistically significant (p>.25). The semen volume was significantly increased in male infertility patients and fertility group (p<.05), but there was no statistical significant difference in impotent patients (p>.25). The sperm count was improved in male infertility patients and fertility group, but this improvement was not statistically significant (p>.25), and in impotent patients there was no significant difference (p>.40). The sperm motility was very significantly increased in male infertility patients and the fertility group (p<.0005), and highly significantly increased in impotent patients (p<.005). The abnormal forms were not statistically significant in impotence and male infertility patients (p>.40), but significantly decreased in the fertility group (p<.05). Serum testosterone was very significantly increased in impotence and male infertility patients (p<.0005) and was highly significantly increased in fertility groups (p<.005). Serum FSH was improved in impotence and male infertility patients, but this improvement was not statistically significant (p>.10), and in fertility groups of male infertility patients, the results showed a statistically significant increase (p<.05). Serum LH was not statistically significant in impotence and male infertility patients (p>.10), and was significantly increased in fertility groups (p<.05). The improvement of sexual activity was 50-75%, the pregnancy rate for their partners was 37% and increased plasma testosterone levels over a period of 3 years of follow-up after varicocele repair. PMID- 12746102 TI - Effect of tyrosine kinase inhibitors on tyrosine phosphorylation and motility parameters in human sperm. AB - Tyrosine phosphorylation has recently been associated with capacitation and suggested as a regulator of sperm movement, especially characterizing hyperactivation. The objective of this study was to verify if tyrosine phosphorylation of human sperm proteins was essentially required for the maintenance of motility as well as the development of hyperactivation. Washed sperm were incubated for 6 h in Ham's F10 + 0.35% HSA at 37 degrees C in 5% CO(2), with and without the tyrosine kinase inhibitors genistein, tyrphostin, erbstatin, or herbimycin A and the wide-spectrum kinase inhibitor staurosporin. The concentrations of the inhibitors used in the experiments did not induce sperm toxicity, as measured by membrane integrity and mitochondrial function assays. Samples incubated without the inhibitors (control), increased their tyrosine kinase activity (ELISA), the number and intensity of tyrosine-phosphorylated (PY) protein bands (Western blot), the incidence of PY-immunoreactive sperm (immunofluorescence), and some of the sperm motion characteristics (CASA), such as velocity (VEL), amplitude of lateral head displacement (ALH), and hyperactivation. Among the selective protein tyrosine kinase inhibitors, genistein was the most active and consistent, inhibiting sperm tyrosine kinase activity, PY proteins, incidence of PY sperm, and sperm motility and motion parameters, such as VEL, ALH, and hyperactivation. The rest of the kinase inhibitors decreased motion characteristics to a varied extent and had different effects on phosphorylation parameters. In general, they decreased PY phosphorylation of 2 proteins (83 and 54 kDa) present in whole sperm extracts, and two sets of proteins of low (39-49 kDa) and medium (55-87 kDa) molecular weight present in the Triton X-100-solubilized sperm protein fraction. This inhibition was evident regardless of the total tyrosine kinase activity of the samples or the incidence of PY-immunoreactive sperm. The described findings further support the association between motility and protein tyrosine phosphorylation in human sperm and point to certain proteins as the main linkers. PMID- 12746103 TI - Constitutive and inducible expression of CYP enzymes in immortal hepatocytes derived from SV40 transgenic mice. AB - 1. The expression of liver-specific transcription factors and cytochrome P450 (CYP) enzymes have been studied in three new hepatocyte-like cell lines derived from SV Delta 202 transgenic mice: AMH-Delta 202 (adult mouse hepatocytes), TAMH Delta 202 (tumour-derived adult mouse hepatocytes) and NMH-Delta 202 (newborn mouse hepatocytes). 2. mRNA levels of liver-enriched transcription factors such as D-element binding protein (DBP), liver-enriched transcription activating protein (LAP) and the hepatic nuclear factors (HNF) 1, 2 and 3 in all Delta 202 transgenic hepatocyte lines were similar to those in the wild-type liver and in primary mouse hepatocytes. 3. Analysis of basal CYP activities and testosterone metabolism revealed that Delta 202 cells showed higher similarities to mouse hepatocytes than Hepa 1c1c7 hepatoma cells. All three Delta 202 cell lines exhibited substantial active CYP1A1/2, CYP2A4/5 and CYP3A11 activities and lower levels of CYP2B, CYP2C and CYP2E1 activities. 4. The Delta 202 cells also responded to model inducers. 3-Methylcholanthrene induced CYP1A1/2 (7 ethoxyresorufin O-deethylation); phenobarbital induced CYP2B (7-benzoxyresorufin O-debenzylation), CYP2A4/5 (testosterone 7alpha -hydroxylation) and CYP3A11 (testosterone 6beta -hydroxylation); and rifampicin and dexamethasone induced CYP3A11 activities in the three Delta 202 cell lines, whereas only AMH-Delta 202 cells reproduced to a limited extent the response of CYP2E1 to ethanol observed in hepatocytes. 5. The results suggest that generation of hepatocyte lines from transgenic animals constitutes a successful approach to obtain in vitro models alternative to primary hepatocytes for drug metabolism and CYP inducibility studies. PMID- 12746104 TI - Prediction of in vivo hepatic clearance from in vitro data using cryopreserved human hepatocytes. AB - 1. Cryopreserved human hepatocytes were used to predict in vivo hepatic clearance (CL(hepatic)) from estimates of in vitro intrinsic clearance (CL' int). 2. (CL' int) was estimated for phenytoin, valproic acid, carbamazepine, theophylline, quinidine and procainamide after their addition to hepatocytes suspended either in human serum or in serum-free media. (CL' int)was estimated from in vitro concentration versus time data fitted to a monoexponential decay model. (CL' int) was estimated from concentrations measured at four time points and from just two point measures, namely the initial concentration (C(0)) and the final concentration measurement (C(last)). 3. Predicted CL(hepatic) was within twofold of reported in vivo values of CL(hepatic) for all substrates. Moreover, predictions were not significantly different whether derived from hepatocytes suspended in serum or in serum-free medium. 4. Two-point estimates of (CL' int) were just as accurate in predicting CL(hepatic) as were multipoint estimates of (CL' int). 5. Although the data set was limited, the findings suggest that the measurement of the disappearance of xenobiotics from serum or serum-free media in which primary human hepatocytes have been suspended provides a physiologically relevant estimate of hepatic clearance that can be employed early in the drug development process to eliminate xenobiotics with unacceptable clearances. PMID- 12746105 TI - Phase II in vitro metabolism of 3-methylindole metabolites in porcine liver. AB - 1. The Phase II in vitro metabolism of 3-methylindole (3MI) metabolites was investigated in pigs to determine the possible relationship between 3MI Phase II metabolism and 3MI accumulation in fat. Sulphation and glucuronidation of five of the seven major metabolites found to be produced by porcine microsomes was investigated using porcine cytosol and microsomes, respectively. The possible formation of glutathione conjugates was also investigated using microsomally activated 3MI intermediate(s). 2. No sulphation or glucuronidation was observed for metabolites 3-hydroxy-3-methyloxindole, 3-methyloxindole, indole-3-carbinol or 2-aminoacetophenone; however, 5-hydroxy-3-methylindole (5-OH-3MI) was conjugated with both sulphate and glucuronic acid. 3. The enzyme responsible for sulphation of 5-OH-3MI was identified as the thermostable form of phenol sulphotransferase (TS-PST) based on its susceptibility to TS-PST inhibitors and the correlation between sulphation of 5-OH-3MI and sulphation of the prototype substrate p-nitrophenol (r = 0.94, p < 0.001). 4. A 3MI-glutathione adduct was identified in microsomal incubations containing 3MI and glutathione. 5. Sulphation of 5-OH-3MI was high in pigs with low levels of 3MI in fat. No relationship was observed between 3MI levels in fat and either glutathione transferase or glucuronidation activities in liver. PMID- 12746106 TI - Identification of novel metabolites of pioglitazone in rat and dog. AB - 1. Four new metabolites of pioglitazone were identified by liquid chromatography mass spectrometry (LC-MS/MS) as being formed by hydroxylation (M-VII and M-VIII), opening of the thiazolidinedione ring (M-X) and by desaturation of the terminal ethyl side chain or tether ethoxy moiety (M-IX), respectively. The structure of one of the hydroxylated metabolites (M-VII) was confirmed by chemical modification using the Jones reaction. 2. Oxidative cleavage of the thiazolidinedione ring is a novel pathway not previously reported for pioglitazone. 3. The hydroxylated M-VII was detected in incubations with rat, dog and human liver and kidney microsomes, and in plasma from rats and dogs dosed orally with [(3)H]pioglitazone. 4. The carboxylic acid derivative of M-VII (M-V) and its taurine conjugate were the major radioactive components in dog bile. PMID- 12746107 TI - Studies on the induction of rat hepatic CYP1A, CYP2B, CYP3A and CYP4A subfamily form mRNAs in vivo and in vitro using precision-cut rat liver slices. AB - 1. Real-time quantitative reverse transcription-polymerase chain reaction methodology (TaqMan(R)) was used to examine the induction of some selected rat hepatic cyto-chrome P450 (CYP) forms in vivo and in vitro using cultured precision-cut liver slices. 2. TaqMan primers and probe sets were developed for rat CYP1A1, CYP1A2, CYP2B1, CYP2B1/2, CYP3A1, CYP3A2 and CYP4A1 mRNAs. 3. To characterize the responsiveness of the rat CYP mRNA TaqMan primers and probe sets, rats were treated in vivo with a single intraperitoneal dose of 500 mg kg( 1) Aroclor 1254 (ARO) and with four daily oral doses of either 50 mg kg(-1) day( 1) dexamethasone (DEX) or 75 mg kg(-1) day(-1) methylclofenapate (MCP). Treatment with ARO produced 22 600-, 5480-, 648-, 52-, 47- and 9-fold increases in levels of CYP1A1, CYP2B1, CYP2B1/2, CYP1A2, CYP3A1 and CYP3A2 mRNA, respectively. DEX treatment produced 97-, 24-, 8- and 4-fold increases, respectively, in CYP3A1, CYP2B1, CYP2B1/2 and CYP3A2 mRNA levels, and MCP produced 339-, 126- and 25-fold increases, respectively, in CYP4A1, CYP2B1 and CYP2B1/2 mRNA levels. All three CYP inducers also increased microsomal CYP content and produced corresponding increases in CYP1A, CYP2B, CYP3A and CYP4A form marker enzyme activities. 4. Rat liver slices were cultured for 6 and 24 h in medium containing 0.1 micro M insulin and 0.1 micro M DEX, and also for 24 h in medium containing only 0.1 micro M insulin (DEX-free medium). Liver slices were cultured in control medium or in medium containing either 10 micro M beta-naphthoflavone (BNF), 10 micro g ml(-1) ARO, 500 micro M sodium phenobarbitone (NaPB), 20 micro M pregnenolone 16alpha -carbonitrile (PCN), 50 micro M Wy-14,643 (WY) or 50 micro M MCP. 5. With the exception of the effect of BNF on CYP1A1 mRNA levels, the induction of all the CYP mRNAs studied was greater after 24- than after 6-h treatment. Generally, the magnitude of induction of CYP mRNA levels was greater after 24 h in liver slices cultured in DEX-free than in DEX-supplemented medium. 6. Treatment of liver slices with BNF and ARO for 24 h in DEX-free medium produced 21- and 35 fold increases, respectively, and 38- and 37-fold increases, respectively, in CYP1A1 and CYP1A2 mRNA levels. NaPB, PCN, WY and MCP did not increase either CYP1A1 or CYP1A2 mRNA levels. 7. After 24 h, levels of CYP2B1/2 mRNA were increased 18-, 20-, 9-, 16- and 13-fold by treatment with ARO, NaPB, PCN, WY and MCP, respectively. PCN also produced 56- and 4-fold increases, respectively, in CYP3A1 and CYP3A2 mRNA levels. 8. Treatment with WY and MCP for 24 h produced 437 and 186-fold increases, respectively, in levels of CYP4A1 mRNA. None of the other CYP inducers studied had any effect on CYP4A1 mRNA levels. 9. The results demonstrate the utility of cultured precision-cut liver slices as an in vitro model system to evaluate the effects of xenobiotics on rat CYP1A, CYP2B, CYP3A and CYP4A form mRNA levels. PMID- 12746108 TI - More potent inhibition of human CYP2A6 than mouse CYP2A5 enzyme activities by derivatives of phenylethylamine and benzaldehyde. AB - 1. A rapid 96-well plate assay method was developed and validated to measure liver microsomal coumarin 7-hydroxylation in vitro. 2. The method was used to test inhibition of human and mouse CYP2A enzymes by three phenylethylamine derivatives 2-(p-tolyl)-ethylamine, amphetamine, 2-phenylethylamine and benzaldehyde, and two of its derivatives, 4-methylbenzaldehyde and 4 methoxybenzaldehyde. 3. The benzaldehyde derivatives were more potent inhibitors of CYP2A5 than the phenylethylamines. The K(ic) value of 4-methylbenzaldehyde was 3.4 micro M and for 4-methoxybenzaldehyde it was 0.86 micro M for CYP2A5. 4. Amphetamine is a weak inhibitor of CYP2A6, whereas benzaldehyde is a suicide inhibitor with K(inact) = 0.16 min(-1) and K(I) = 18 micro M. The K(ic) values of 2-phenylethylamine, 2-(p-tolyl)-ethylamine, 4-methylbenzaldehyde and 4 methoxybenzaldehyde were 1.13, 0.23, 0.36 and 0.73 micro M for CYP2A6, respectively. 5. Novel potent inhibitors were found for CYP2A6 and, except for 4 methoxybenzaldehyde, all the compounds inhibited CYP2A5 and CYP2A6 enzymes differentially. These data add to the refinement of CYP2A enzyme active sites and provide chemical leads for developing novel chemical inhibitors of the CYP2A6 enzyme. PMID- 12746109 TI - Pharmacokinetics and disposition of a novel NMDA glycine site antagonist (UK 240,455) in rats, dogs and man. AB - 1. UK-240,455 ((+) 6,7-dichloro-5-[N-(2-hydroxyethyl)methanesulphonamido]-2,3 (1H,4H)-quinoxalinedione) is a potent, selective N-methyl D-aspartate (NMDA) glycine site antagonist that is being evaluated for the potential treatment of stroke. 2. UK-240,455 is predominately excreted unchanged in urine (58-68%) in rats, dogs and man following intravenous administration. The remainder of the dose is excreted unchanged in the faeces. It is considered that UK-240,455 is predominantly cleared by active renal tubular secretion and active hepatobiliary transport. In man, there is evidence that UK-240,455 undergoes glucuronidation. However, there is no evidence for this in rats and dogs. 3. UK-240,455 has a short elimination half-life in rats, dogs and man (0.4-1.4 h) and clearances of 12, 13 and 6 ml min(-1) kg(-1), respectively. The compound shows limited tissue distribution with volumes of distribution of 0.4-0.8 l kg(-1) in rats, dogs and man. The species' variation in pharmacokinetic parameters was related allometrically when plasma protein binding was taken in to account. Hence, active hepatic or renal clearance processes for this compound were conserved across species. 4. Cerebrospinal fluid and brain concentrations of UK-240,455 were determined in rats. The cerebrospinal fluid/plasma concentration ratio of UK 240,455 was 4.3%, which was similar to the plasma-free fraction of this compound (3%), indicating good blood-brain barrier permeability. Brain tissue concentrations were low (0.7% of the total plasma concentrations). PMID- 12746112 TI - Thresholds in chemical carcinogenesis: what are animal experiments telling us? AB - It appears that the controversy over whether animal experiments demonstrate a threshold for carcinogenicity from chemicals was due to an error in plotting dose response. A linear (arithmetic) scale for the dose of chemicals obscures effects at doses below those used in the experiment and distorts the effect seen over the range of doses used. Gaddum (Nature 156: 463, 1946) pointed out that, empirically, dose should be on a logarithmic scale to effect a linear quantal response. It now is proposed that this logarithmic relationship of dose to effect has a sound basis in chemical thermodynamics (Waddell, Toxicol Sci 68: 275-279, 2002). When the results of major studies (e.g., ED01, etc.) done in the past were reanalyzed with this in mind using the Rozman et al scale (Drug Metab Rev 28: 29 52, 1996), which has a logarithmic scale down to one molecule (10(0)), unequivocal thresholds were demonstrated. Many animal studies now have been reanalyzed by this procedure (e.g., Waddell, Toxicol Sci 72: 158-163, 2003; Waddell, Food Chem Toxicol, in press; Waddell, Hum Exp Toxicol, in press); these studies show thresholds for carcinogenicity at doses ranging from 10(17.1) to 10(21.92) molecules/kg/day. These results require a complete reevaluation of human risk assessment for carcinogenesis from chemicals and a redirection of basic research to discover the system, or systems, that are overwhelmed at these thresholds. PMID- 12746110 TI - Identification of coenzyme A-related tolmetin metabolites in rats: relationship with reactive drug metabolites. AB - 1. It has recently been proposed that acyl coenzyme A thioesters (acyl-CoAs) of xenobiotic carboxylic acids are electrophilic, reactive metabolites that may react with proteins. 2. The primary objective was to investigate the reactivity of the tolmetin acyl coenzyme A thioester (Tol-CoA). The second objective was to identify and quantify tolmetin (Tol) metabolites in vivo that were formed via Tol CoA, e.g. the glycine (Tol-Gly) and taurine (Tol-Tau) conjugates. This finding would be indicative of Tol-CoA formation and thus of other acyl-CoA-related reactions that might occur, e.g. covalent binding to proteins. 3. In order to study the chemical reactivity, Tol-CoA (0.5 mM) was incubated with glutathione (5 mM) in a 0.1 M phosphate buffer (pH 7.4) at 37 degrees C. Tol-CoA reacted rapidly with glutathione in vitro to form the S-acyl glutathione conjugate at a rate of 14.9 +/- 0.7 micro M min(-1) (mean +/- SD, n = 3) from 0 to 10 min. Compared with acyl-CoAs of other xenobiotic carboxylic acids, naproxen and clofibric acid, the rate by which Tol-CoA reacted with glutathione was high. 4. Following administration of (3)H-Tol (100 mg kg(-1), 200 micro Ci kg(-1), p.o.) to male Sprague-Dawley rats, Tol-Tau and Tol-Gly were identified in urine by electrospray ionization MS-MS in both positive- and negative-ion modes. The conjugates were only formed at trace levels (< 0.5%). However, the presence of Tol-Tau and Tol Gly showed the reactive Tol-CoA was formed in vivo. PMID- 12746113 TI - Endothelin antagonist-induced coronary and systemic arteritis in the beagle dog. AB - Two endothelin antagonists, ZD1611 (3-[4-[3-(3-methoxy-5-methylpyrazin-2 ylsulfamoyl)-2-pyridyl]phenyl]-2,2-dimethylpropanoic acid) and ZD2574 (2-(4 isobutylphenyl)-N-(3-methoxy-5-methylpyrazin-2-yl)pyridine-3-sulfonamide), selective for the ET(A) receptor and intended for use in pulmonary hypertension, were tested in Beagle dogs at various doses for periods of up to 4 weeks. These studies included in vivo telemetric hemodynamic assessment, full histopathological and ultrastructural pathological evaluation of coronary arteries. Both drugs produced arteritis in small- and medium-sized coronary arteries after single or multiple doses, some of which were at or below the ED50. The distribution of lesions was predominantly in extramural arteries over the atria and atrioventricular groove of the right side of the heart and consisted of epicardial hemorrhage and arteritis. Systemic arteritis was also present at a lower incidence than the coronary arteritis, was located at different sites and appeared inconsistently. Ultrastructural changes in coronary arteries suggested that damage was the result of mechanical factors. Although these patterns of vascular injury possessed features in common with those induced in dogs by high doses of vasodilating antihypertensive drugs and inotropic agents, they were atypical, as there was no left ventricular myocardial necrosis, papillary muscle damage, or subendocardial hemorrhage suggestive of ischaemia or excessive inotropism. Moreover, physiological monitoring showed no evidence of exaggerated systemic hypotension or reflex tachycardia at doses associated with vascular damage. Consequently, the changes might be the result of a localized pharmacological process such as intense, prolonged vasodilatation in unsupported arteries that are well endowed with endothelin receptors and particularly sensitive to endothelin antagonism. PMID- 12746114 TI - Main vascular changes seen in the saline controls of continuous infusion studies in the cynomolgus monkey over an eight-year period. AB - The aim of this study was to characterize the main vascular lesions seen in the saline controls in continuous infusion studies in monkeys. Data were collected from 17 studies over an 8-year period (ie, around 50 males and 50 females). The study was restricted to lesions at the injection site, in lungs and at the entry point. It showed that the level of the most severe procedure-related lesions (ie, necrosis with abscess formation at the injection site and at the entry point, thromboemboli and probable infarcts in lungs) was low (<10%). Minor changes such as endothelial hyperplasia and intimal thickening at the injection site, and low graded interstitial pneumonitis in lungs were the most frequent changes (40 to 50% of the animals). Thrombi at the injection site were present in around 30% in males and 40% in females. The slightly higher incidence of thrombi in females was not explained by a difference of vessel size or by hematological differences. This study permitted an opportunity to harmonise terminology among pathologists and to define the main procedure-related changes and their incidence, which could help pathologists better interpret changes in future infusion studies. PMID- 12746115 TI - Development of sperm granulomas in the epididymides of L-cysteine-treated rats. AB - We investigated the histopathological effects of excess L-cysteine on the male rat reproductive tract during sexual maturation. Male 6-week-old Sprague-Dawley rats were injected intraperitoneally daily with L-cysteine, 1,000 mg/kg body weight, for 1, 2, 3, and 4 weeks. L-Cysteine-treated rats developed sperm granulomas in the epididymides at an incidence of 0% (0/6), 50% (3/6), 83% (5/6), and 100% (6/6) in rats examined at study weeks 1, 2, 3, and 4, respectively. These sperm granulomas were unilateral or bilateral, and most frequently involved the proximal cauda region of the epididymides. Interestingly, small ducts, indicative of immaturity, were seen frequently in L-cysteine-treated rats. These findings suggest that the maturation of epididymides in L-cysteine-treated rats might be delayed. Additionally, dilated ducts and interstitial edema, suggestive of an increase in intraluminal pressure, were seen often in the epididymides of L cysteine-treated rats. Labeling spermatozoa and epithelial cells with monobromobimane indicated no influence of the thiol-disulfide status of L cysteine to the epididymides. The testes and prostate glands also showed no effects, suggesting that inhibited epididymis maturation was not a result of hormonal deficiencies. We speculate that defective development of the ducts might result in aberrant fluid flow, leading to ductal rupture in the epididymides. In that case, sperm granulomas might form around leaked spermatozoa. PMID- 12746116 TI - Case report of rat true hermaphroditism: colocalization of oocytes and granulosa and sertoli cells in the germinal cord. AB - We describe a case of rat hermaphroditism with bilateral ovotestes. In a 7-week old apparently male Sprague-Dawley rat, both testes were relatively small, and the right testis with a faint protrusion was somewhat round and small as compared with the left testis. Microscopically, the testes contained ovarian tissues within their tunica albugineas in conjunction with spermatogenesis in the seminiferous tubules. As bilateral changes, oocytes surrounded by granulosa-like cells were present in the seminiferous tubule-like germinal cord. Granulosa-like and Sertoli-like cells were layered together on the basal lamina, and theca interna-like cells were occasionally observed around the basal lamina. As unilateral changes, cystic dilatation of the germinal cords with eosinophilic fluid was seen in the lumen, and the theca interna-like cells appeared to be vacuolated. Immunohistochemically, the granulosa-like and Sertoli-like cells showed positive reactions for 3beta-hydroxysteroid dehydrogenase (3beta-HSD) and vimentin, respectively. Theca interna-like cells reacted positively to both 3beta HSD and cytochrome P-450 17alpha-hydroxylase. Ultrastructurally, the granulosa, Sertoli, and theca interna cells were also identified in the ovarian tissue. From these morphological characteristics, the male rat with bilateral ovotestes was diagnosed as true hermaphroditism. PMID- 12746117 TI - Microarray analysis using amplified mRNA from laser capture microdissection of microscopic hepatocellular precancerous lesions and frozen hepatocellular carcinomas reveals unique and consistent gene expression profiles. AB - The indirect labeling cDNA microarray technique was used to evaluate gene expression profiles of pure cell populations from frozen sections of carcinomas and adenomas harvested from precancerous hepatocellular lesions by using laser capture microdissection (LCM). The levels of differentially expressed genes were investigated using a cDNA microarray with 9,984 features with only 2 ug of two round amplified aRNA, equivalent to 35 cells from LCM-adenomas and frozen samples of carcinomas from simian virus 40 (SV40) large T antigen transgenic rats. A total of 855 genes were identified as being 3-fold or more differentially expressed in carcinomas or adenomas as compared to normal tissue controls. Among these 855 genes, 71 genes were differentially expressed in both carcinomas and adenomas. Commonly up-regulated genes in both carcinoma and adenomas were 28 while 41 of the 71 genes were commonly down-regulated. Two genes, Igh1 (immunoglobulin heavy chain 1(Serum IgG2a), Image clone ID: 875880) and EST clone (AI893585, Image clone ID: 596604) were more than 7-fold up-regulated in carcinomas and 6-fold down-regulated in adenomas. In Cy5 and Cy3 reciprocal experiments for screening out false positive signals, the amplified carcinomas showed higher Pearson Correlation Coefficient values (-0.94 and -0.92) than the LCM-amplified adenoma samples (-0.79 and -0.84). LCM-amplified samples provided higher signal intensities over backgrounds and a greater average of Cy5:Cy3 ratios. Expression levels of mRNAs from selected genes, determined by using traditional dot blot analysis, revealed that 36 of 40 tested expression profiles were consistent with the microarray data. Thus, amplified aRNA harvested from homogeneous cell types using LCM can be applied to study gene expression profiles by use of microarray analysis. PMID- 12746118 TI - Histiocytic sarcoma in C57BL/6J female mice is associated with liver hematopoiesis: review of 41 cases. AB - Forty-one cases of histiocytic sarcoma (HS) in C57BL/6J mice were histopathologically studied with special regard to unexpected associated hematopoietic disorders. These cases were retrieved among C57BL/6J female mice used as control mice in a chronic low-dose irradiation experiment. Hematopoietic characteristics were analysed by comparison to 41 disease-free mice from the same cohort. Tumoral involvement of the liver was observed in all 41 HS-bearing mice, followed by infiltration of the spleen (61.8%), lung (32.4%), bone marrow (14.3%), uterus (12.2%), lymph node (9.8%), and kidney (2.4%). By comparative analysis, we were able to demonstrate a significant association of HS with liver hematopoiesis (89.5% in HS group vs 15% in control mice, p < 0.00001), and with central hematopoietic disorders involving the myelocytic cells (decreased in HS, p = 0.003) and erythrocytic cells (increased in HS, p = 0.001). Microscopic characteristics of these 41 cases and physiopathology of the newly described hematopoietic features in HS are further discussed. PMID- 12746120 TI - Hylan G-F 20 induces delayed foreign body inflammation in Guinea pigs and rabbits. AB - Recent clinical evidence suggests that hylan, a modified hyaluronan, and related products potentially elicit foreign body granulomatous inflammation in human soft tissue. We investigated the biocompatibility of hylan G-F 20 (Synvisc) for up to 28 days after intradermal injection in guinea pigs and intramuscular injection in rabbits. Compared to saline and unmodified hyaluronan, hylan induced definitive macroscopic changes in guinea pigs by day 14 or later and in rabbits by 28 days after injection. Histologically, at the injection sites, there was severe granulomatous inflammation in guinea pigs and acute inflammation with minimal infiltration of macrophages and foreign body giant cells in rabbits. Furthermore, specific antibodies against hylan were demonstrated in guinea pigs by passive cutaneous anaphylaxis, and substantial deposits of IgG on hylan were evident by immunohistochemistry. The present results contradict previous reports on biocompatibility of hylan and suggest that hylan may potentially induce similar unfavorable reactions in humans. PMID- 12746119 TI - The effects of ad libitum overfeeding and moderate and marked dietary restriction on age-related spontaneous pituitary gland pathology in Sprague-Dawley rats. AB - This study compared the effects of ad libitum (AL) overfeeding and moderate or marked dietary restriction (DR) on the pathogenesis of aged-related pituitary gland changes in Sprague-Dawley (SD) rats. SD rats were fed Purina Certified Rodent Diet AL (group 1), DR at 72-79% of AL (group 2), DR at 68-72% of AL (group 3) or DR at 47-48% of AL (group 4) for 106 weeks. Interim necropsies were performed at 13, 26, and 53 weeks, after a 7-day 5-bromo-2-deoxyuridine (BrdU) filled minipump implantation. Body weights, organ weights and insulin-like growth factor 1 (IGF-1) serum levels were measured at interim and final necropsies. Serum levels of prolactin (PRL), progesterone, estradiol, luteinizing hormone (LH) and follicle stimulating hormone (FSH) were measured at 53 and/or 106 weeks. In addition to the routine histopathologic examination, determination of 7 stereologic parameters after pituitary immunohistochemistry of PRL, growth hormone (GH) and BrdU was done in both sexes at 13, 26, and 53 weeks. Body and pituitary weights were proportional to the food intake. In AL-fed rats, hyperplastic and neoplastic changes developed early and progressed with age, affecting almost all animals by 106 weeks. These changes were associated with high PRL serum levels. Pituitary adenomas were the most common cause of death in both sexes. In DR rats, a delayed onset and a decreased incidence of pituitary tumors were observed in association with decreased serum IGF-1, PRL, estradiol, and LH levels. The results of the stereological analysis demonstrated that, compared to AL-fed rats, pituitary glands from DR rats contained lower PRL and GH secreting cell volumes, and a lower epithelial cell BrdU labeling index, which correlated with a lower incidence of pituitary tumors at study termination. Moderate and marked degrees of DR delayed the onset of pituitary tumors in a temporal- and dose-related manner. In contrast to marked DR, which dramatically reduced the incidence of hyperplastic and neoplastic pituitary gland changes, moderate DR delayed the onset but did not prevent the development of pituitary tumors. PMID- 12746121 TI - Toxic rhinitis-induced changes of human nasal mucosa innervation. AB - Irritative toxic rhinitis is a nasal disorder induced by chemical compounds like ozone, formaldehyde, nickel, chrome, solvents and tobacco smoke. These noxious stimuli may have effects on the nasal innervation leading to a cascade of neuro immune interactions and an augmentation of the symptoms. Here we examined changes in the neuropeptide content of mucosal parasympathetic, sympathetic and sensory nerves of patients with toxic rhinitis caused by chronic cigarette smoke exposure. Semiquantitative immunohistochemistry using antibodies against calcitonin gene-related peptide (CGRP), substance P (SP), neuropeptide tyrosine (NPY), and vasoactive intestinal peptide (VIP) was carried out on cryostat sections of human nasal mucosa obtained from normal subjects and patients with toxic rhinitis and revealed significant differences between both groups. Toxic rhinitis patients had significantly elevated expression scores for VIP (2.83 +/- 0.31 vs 1.27 +/- 0.47 control group) and NPY (3.17 +/- 0.31 vs 0.91 +/- 0.37 control group) revealing an increase of mediators in distinct subpopulations of airway nerves. In summary, the present studies indicate a differential participation of subclasses of mucosal nerves in the pathophysiology of toxic rhinitis. Airway innervation may have a major role in the pathophysiology of toxic rhinitis associated with chronic cigarette smoke exposure. PMID- 12746122 TI - Development of methodology for the three-dimensional modelling of the metabolic capacity of the rat nasal cavity using glutathione S-transferase M1 as an example. AB - A variety of chemicals induce site-specific lesions in the rodent nasal cavity. In order to explore the reasons for this site-selectivity, methodology for (a) creation of a 3-dimensional (3D) model of a rat nasal cavity, and (b) mapping of semiquantitative data onto the model has been developed. The head of a rat was fixed, decalcified, step-sectioned (every 100 microm) and stained with hematoxylin and eosin. Digital images of the sections were optically captured, and a KS400 image analysis system (Imaging Associates, Thame, Oxford, UK), attached to a standard personal computer, was used to align adjacent images and reconstruct the series in 3D. The final model was anatomically correct, and could be rotated in any plane and manipulated to display individual internal structures. The spatial localization of a glutathione S-transferase (rGSTM1, previously known as GST 3-3) within this model was investigated using immunohistochemistry. Step sections (every 400 microm) were stained, analyzed by imaging densitometry, and the results for the stained regions within the nasal cavity divided into 4 grades representing high to low expression of rGSTM1. The data was mapped onto the 3D model and showed that the highest expression of this enzyme was in the central regions of the nasal cavity at the transition between respiratory and olfactory epithelia. This methodology will allow investigation of the relationship between the in situ localization of bioactivating and detoxifying enzyme systems and the site-specificity of nasal lesions. PMID- 12746123 TI - Three-dimensional mapping of the lesions induced by beta-beta' iminodiproprionitrile, methyl iodide and methyl methacrylate in the rat nasal cavity. AB - The nasal cavity is an important target organ for toxicity, and many chemicals induce site-specific lesions in this region. The factors responsible for this site-selectivity have not been unequivocally identified, but probably include regional dosimetry and bioactivation. The purpose of this study was to map, in 3 dimensions, the lesions induced by beta-beta'-iminodipropionitrile (IDPN), methyl iodide (MeI) and methyl methacrylate (MMA) in the rat nasal cavity. Animals were administered IDPN (150 mg/kg, IP) or exposed via inhalation to MeI (100 ppm, 2 hours) or MMA (400 ppm, 4 hours) and sacrificed after 24 hours. Heads were decalcified, step-sections (1 every 400 microm) cut and stained, and the severity of the epithelial lesion graded as mild (vacuolation and pyknosis), moderate (undulation and mild stripping), or marked (complete stripping). These grades were mapped onto a 3D-model of a rat nasal cavity using the KS400 imaging system (Imaging Associates, Thame, UK). Despite the different routes of exposure the lesions induced by the 3 compounds had very similar distributions, predominantly affecting the dorsal-medial aspects of the ethmoturbinates and, in the case of MMA, the organ of Rodolfo Masera. These results suggest that, with these chemicals, local bioactivation plays a more important role than dosimetry in determining lesion distribution. PMID- 12746124 TI - Food additives and cancer: banging the drum. PMID- 12746126 TI - Biomarkers of renal effects in children and adults with low environmental exposure to heavy metals. AB - The health effects of chronic exposure to heavy metals such as lead, cadmium, and mercury are widely documented, yet few data exist about the renal impact of low environmental exposure to these metals, particularly in children. The aim of this study was to assess renal parameters in children and adults living in an environment known for its past heavy metal contamination around two nonferrous smelters in northern France (Noyelles-Godault and Auby) and to compare their results with age and gender-matched controls living in neighboring municipalities with unpolluted soil (total: 400 children, 600 adults, sex ratio = 1). The integrity of renal function was assessed by measuring the urinary excretion levels of total protein, albumin, transferrin, beta(2)-microglobulin, retinol binding protein, brush border antigen, and the enzyme N-acetyl-beta-D glucosaminidase (NAG). The mean blood concentrations of lead (Pb-B, children 1 suggest the potential to produce OPIDN at doses lower than the LD50. Inhibition of NTE and AChE in hen brain in vivo was studied 24 h after i.m. injection of hens with increasing doses of methyl and butyl derivatives. Analysis of dose-response curves yielded ED50(AChE)/ED50(NTE) ratio of 0.86 for methyl PhP and 22.1 for butyl PhP. These results predict that the butyl derivative should be more neuropathic than the methyl analogue. Excellent correspondence between in vivo and in vitro predictions of neuropathic potential indicate that valid predictive QSAR models may be based on the in vitro approach. Adoption of this system would result in reducing experimental animal use, lowering costs, accelerating data production, and enabling standardization of a biochemically based risk assessment of the neuropathic potential of OP compounds. PMID- 12746137 TI - Effect of a chemical mixture on dermal penetration of arsenic and nickel in male pig in vitro. AB - The effect of a chemical mixture on the dermal penetration of arsenic or nickel was assessed by applying arsenic-73 or nickel-63 alone or with the chemical mixture to dermatomed male pig skin samples in flow-through diffusion cells. The chemical mixture consisted of chloroform, phenanthrene, and toluene for arsenic penetration studies and phenol, toluene, and trichloroethylene (TCE) for nickel studies. These are predominant chemicals found at hazardous waste sites. Arsenic and nickel bind to skin after dermal exposure. Total penetration of arsenic and nickel in the chemical mixture were significantly increased by 33% and 20% compared to arsenic and nickel alone, respectively. While more radioactivity penetrated skin with chemical treatment than metal alone, significantly less radioactivity was loosely adsorbed to skin and could be easily washed off from the skin surface with soap and water. The results of this study indicate that the potential health risk from dermal exposure to arsenic or nickel is enhanced if other chemicals are present. PMID- 12746138 TI - Hormonal, morphological, and physiological responses of yellow perch (Perca flavescens) to chronic environmental metal exposures. AB - The effects of a chronic environmental exposure to metals on the hormonal, physiological, and reproductive status were assessed in yellow perch (Perca flavescens) sampled in six lakes situated along a contamination gradient of Cd, Zn, Cu, Pb, and Ni in the mining region of Rouyn-Noranda, Quebec. Fish were captured in the summer and fall, and sampled before or after a confinement of one hour. Metal concentrations in the kidneys and the interrenal tissues (homologous to mammalian adrenals) were measured to compare tissue-specific metal accumulation. An exposure-related decrease of condition factor, gonadosomatic index (GSI), branchial Na+/K(+)-ATPase activity, plasma thyroxine (T4), triiodothyronine (T3), and 17 beta-estradiol and an impaired capacity to enhance cortisol levels after confinement were observed. Fish from the metal-contaminated lakes possessed gonads at less mature stages and exhibited structural alterations of their gills, interrenal cells, and thyroid follicle epithelium. A comparison of the morphological, biochemical, and physiological endpoints measured in the present study revealed that plasma concentrations of hormones and parameters of gill function were the most affected by metal contamination. The results of this study indicate that lifelong exposures to sublethal concentrations of metals alter the physiological functions of fish and delay reproduction. PMID- 12746139 TI - Potential adverse health effects of genetically modified crops. AB - Genetically modified crops have the potential to eliminate hunger and starvation in millions of people, especially in developing countries because the genetic modification can produce large amounts of foods that are more nutritious. Large quantities are produced because genetically modified crops are more resistant to pests and drought. They also contain greater amounts of nutrients, such as proteins and vitamins. However, there are concerns about the safety of genetically modified crops. The concerns are that they may contain allergenic substances due to introduction of new genes into crops. Another concern is that genetic engineering often involves the use of antibiotic-resistance genes as "selectable markers" and this could lead to production of antibiotic-resistant bacterial strains that are resistant to available antibiotics. This would create a serious public health problem. The genetically modified crops might contain other toxic substances (such as enhanced amounts of heavy metals) and the crops might not be "substantially equivalent" in genome, proteome, and metabolome compared with unmodified crops. Another concern is that genetically modified crops may be less nutritious; for example, they might contain lower amounts of phytoestrogens, which protect against heart disease and cancer. The review of available literature indicates that the genetically modified crops available in the market that are intended for human consumption are generally safe; their consumption is not associated with serious health problems. However, because of potential for exposure of a large segment of human population to genetically modified foods, more research is needed to ensure that the genetically modified foods are safe for human consumption. PMID- 12746140 TI - Cadmium, lung and prostate cancer: a systematic review of recent epidemiological data. AB - Cadmium (Cd) and its compounds were classified as "carcinogenic to humans (Group 1)" by IARC in 1993. The observation of an increased number of lung cancers in a U.S. cohort of cadmium-exposed workers and the finding of tumors in animals exposed to various cadmium compounds apparently played an important role in this assessment. Since this evaluation, several cohorts of cadmium exposed workers have been updated and some additional data regarding environmental exposure to cadmium and cancer risk have been published. The main purpose of this systematic review was to examine whether inclusion of the studies that were not available for the 1993 evaluation might change the overall assessment of the carcinogenic potential of cadmium compounds. A second objective was to examine whether the recent studies are qualitatively better than the older ones and whether they should receive more weight in this assessment. A third issue was to investigate whether a competing effect between nonmalignant respiratory disease (NMRD) and lung cancer may have affected the results for lung cancer in occupationally exposed cohorts. Overall, considering the results of the most recent studies does not suggest that the effect of cadmium on lung cancer increases with improvement of the study design but points to a lower relative risk in the groups exposed to cadmium in the absence of arsenic and nickel. No evidence was found to support the hypothesis that NMRD represents a competing cause of death reducing the mortality from lung cancer. The association between cadmium exposure and prostate cancer was not confirmed in the latest available updates. Studies in environmentally exposed populations do not indicate an increased relative risk of cancer. PMID- 12746141 TI - Science and transscience in carcinogen risk assessment--the European Union regulatory process for trichloroethylene. AB - This is a study of carcinogen risk assessment of the chlorinated solvent trichloroethylene within the European Union existing substances program and the classification and labeling process. The focus is on the most active and influential participants of this process, namely, those from the United Kingdom, Germany, and Sweden, and from industry. The member state and other experts have different opinions regarding the appropriate classification of trichloroethylene for mutagenicity (no classification or category 3) and carcinogenicity (category 3, 2, or 1). In this article these differences are described, as well as how the primary carcinogenicity and mutagenicity data have been interpreted and evaluated by these participants. It is concluded that underlying the different assessments are disagreements about issues that to some degree lie outside the scope of purely scientific considerations. PMID- 12746142 TI - A role for P-glycoprotein in environmental toxicology. AB - P-Glycoprotein (P-gp) is a transmembrane protein, playing significant roles in the process of drug discovery and development and in pest resistance to pesticides. P-gp affects absorption, disposition, and elimination of different compounds and is mainly expressed in intestines, liver, kidneys, heart, colon, and placenta. The expression of P-gp in the blood-brain barrier (BBB) has been associated with the restricted access of many compounds to the central nervous system. Generated knockout mice by disruption of mdr 1a gene, encoding for P-gp, showed that this protein was expressed in the BBB. The absence or the low levels of P-gp elevated drug concentrations in tissues and decreased drug elimination. P gp is responsible for resistance of cells to agents, particularly the anticancer drugs, by removing these drugs from cells. Increased expression of P-gp is implicated in decreased HIV drug availability at certain intracellular sites. The role of P-gp in affecting efficacy and toxicity of environmental toxicants such as pesticides and heavy metals has not been adequately investigated. Studies showed that P-gp contributes to resistance to pesticides in certain pest species, and to decrease toxicity by removing compounds from cells in mammals. Placental drug-transporting P-gp plays a significant role in limiting the transport of toxicants such as potential teratogens to the fetus. Several in vitro or in vivo assays, including using P-gp knockout or naturally deficient mice, were described for testing P-gp modulators. The role of P-gp following concurrent exposure to more multiple compounds needs further research. P-gp modulators should be carefully used, since some modulators that reverse P-gp efflux action in vitro may lead to alterations of tissue function and increase toxicity of xenobiotics in normal tissues. Recent reports from the pharmaceutical studies on the significance of P-gp as transporters in altering the efficacy and toxicity clearly highlight the need for further research in interaction with environmental toxicants. PMID- 12746143 TI - Ecological risk assessment for aquatic organisms from over-water uses of glyphosate. AB - Although the herbicide glyphosate is most widely used in agriculture, some is used for the control of emergent aquatic weeds in ditches, wetlands, and margins of water bodies, largely as the formulation Rodeo. This article presents an ecological risk assessment (ERA) of glyphosate and some of the recommended surfactants as used in or near aquatic systems. Glyphosate does not bioaccumulate, biomagnify, or persist in a biologically available form in the environment. Its mechanism of action is specific to plants and it is relatively nontoxic to animals. As a commercial product, glyphosate may be formulated with surfactants that increased efficacy but, in some cases, are more toxic to aquatic organisms than the parent material. For this risk assessment, three model exposure scenarios--static or low-flow systems such as ponds, flowing waters such as streams, and systems subjected to tidal flows such as estuaries--were chosen and application rates from 1 to 8 kg glyphosate/ha were modeled. Additional measured exposure data from several field studies were also used. As acute exposures are most likely to occur, acute toxicity data were used as effect measures for the purposes of risk assessment. Toxicity data were obtained from the literature and characterized using probabilistic techniques. Risk assessments based on estimated and measured concentrations of glyphosate that would result from its use for the control of undesirable plants in wetlands and over-water situations showed that the risk to aquatic organisms is negligible or small at application rates less than 4 kg/ha and only slightly greater at application rates of 8 kg/ha. Less is known about the environmental fate and toxicology of the surfactants commonly used in combination with the Rodeo formulation of glyphosate. The surfactants used for this purpose were judged not to be persistent nor bioaccumulative in the environment. Distributional analysis of measured deposition concentrations of LI 700, suggest that this surfactant presents an insignificant acute risk to aquatic organisms. Assuming similar applications rates, significant ecological effects would not be expected from the use of some other surfactants such as Induce or X-77. Risks from the use of glyphosate +MON 0818 (Roundup) were slightly greater than those from glyphosate and surfactants such as LI 700; however, in over-water uses, risks were still considered small. Similar small risks were observed for measured concentrations of glyphosate in surface waters resulting from aerial application of Vision (a formulation equivalent to Roundup) to forestry areas in Canada. Concentrations measured after ground application presented a greater risk, but the data were sparse and the assessment is more uncertain. PMID- 12746144 TI - Stochastic mapping of morphological characters. AB - Many questions in evolutionary biology are best addressed by comparing traits in different species. Often such studies involve mapping characters on phylogenetic trees. Mapping characters on trees allows the nature, number, and timing of the transformations to be identified. The parsimony method is the only method available for mapping morphological characters on phylogenies. Although the parsimony method often makes reasonable reconstructions of the history of a character, it has a number of limitations. These limitations include the inability to consider more than a single change along a branch on a tree and the uncoupling of evolutionary time from amount of character change. We extended a method described by Nielsen (2002, Syst. Biol. 51:729-739) to the mapping of morphological characters under continuous-time Markov models and demonstrate here the utility of the method for mapping characters on trees and for identifying character correlation. PMID- 12746146 TI - The geometry of the marmot (rodentia: sciuridae) mandible: phylogeny and patterns of morphological evolution. AB - Marmots have a prominent role in the study of mammalian social evolution, but only recently has their systematics received the attention it deserves if sociobiological studies are to be placed in a phylogenetic context. Sciurid morphology can be used as model to test the congruence between morphological change and phylogeny because sciurid skeletal characters are considered to be inclined to convergence. However, no morphological study involving all marmot species has ever been undertaken. Geometric morphometric techniques were applied in a comparative study of the marmot mandible. The adults of all 14 living marmot species were compared, and mean mandible shape were used to investigate morphological evolution in the genus Marmota. Three major trends were observed. First, the phylogenetic signal in the variation of landmark geometry, which describes mandible morphology, seems to account for the shape differences at intermediate taxonomic levels. The subgenera Marmota and Petromarmota, recently proposed on the basis of mitochondrial cytochrome b sequence, receive support from mandible morphology. When other sciurid genera were included in the analysis, the monophyly of the genus Marmota and that of the tribe Marmotini (i.e., marmots, prairie dogs, and ground squirrels) was strengthened by the morphological data. Second, the marmotine mandible may have evolved as a mosaic of characters and does not show convergence determined by size similarities. Third, allopatric speciation in peripheral isolates may have acted as a powerful force for modeling shape. This hypothesis is strongly supported by the peculiar mandible of M. vancouverensis and, to a lesser degree, by that of M. olympus, both thought to have originated as isolated populations in Pleistocene ice-free refugia. PMID- 12746147 TI - Molecular and morphological phylogenies of ruminantia and the alternative position of the moschidae. AB - The ruminants constitute the largest group of ungulates, with >190 species, and its distribution is widespread throughout all continents except Australia and Antarctica. Six families are traditionally recognized within the suborder Ruminantia: Antilocapridae (pronghorns), Bovidae (cattle, sheep, and antelopes), Cervidae (deer), Giraffidae (giraffes and okapis), Moschidae (musk deer), and Tragulidae (chevrotains). The interrelationships of the families have been an area of controversy among morphology, palaeontology, and molecular studies, and almost all possible evolutionary scenarios have been proposed in the literature. We analyzed a large DNA data set (5,322 nucleotides) for 23 species including both mitochondrial (cytochrome b, 12S ribosomal RNA (rRNA), and 16S rRNA) and nuclear (kappa-casein, cytochrome P-450, lactoferrin, and alpha-lactalbumin) markers. Our results show that the family Tragulidae occupies a basal position with respect to all other ruminant families, confirming the traditional view that separates Tragulina and Pecora. Within the pecorans, Antilocapridae and Giraffidae emerge first, and the families Bovidae, Moschidae, and Cervidae are allied, with the unexpected placement of Moschus close to bovids rather than to cervids. We used these molecular results to assess the homoplastic evolution of morphological characters within the Ruminantia. A Bayesian relaxed molecular clock approach based on the continuous autocorrelation of evolutionary rates along branches was applied to estimate the divergence ages between the major clades of ruminants. The evolutionary radiation of Pecora occurred at the Early/Late Oligocene transition, and Pecoran families diversified and dispersed rapidly during the Early and Middle Miocene. We propose a biogeographic scenario to explain the extraordinary expansion of this group during the Cenozoic era. PMID- 12746148 TI - Outgroup misplacement and phylogenetic inaccuracy under a molecular clock--a simulation study. AB - We conducted a simulation study of the phylogenetic methods UPGMA, neighbor joining, maximum parsimony, and maximum likelihood for a five-taxon tree under a molecular clock. The parameter space included a small region where maximum parsimony is inconsistent, so we tested inconsistency correction for parsimony and distance correction for neighbor joining. As expected, corrected parsimony was consistent. For these data, maximum likelihood with the clock assumption outperformed each of the other methods tested. The distance-based methods performed marginally better than did maximum parsimony and maximum likelihood without the clock assumption. Data correction was generally detrimental to accuracy, especially for short sequence lengths. We identified another region of the parameter space where, although consistent for a given method, some incorrect trees were each selected with up to twice the frequency of the correct (generating) tree for sequences of bounded length. These incorrect trees are those where the outgroup has been incorrectly placed. In addition to this problem, the placement of the outgroup sequence can have a confounding effect on the ingroup tree, whereby the ingroup is correct when using the ingroup sequences alone, but with the inclusion of the outgroup the ingroup tree becomes incorrect. PMID- 12746145 TI - Sampling strategies for delimiting species: genes, individuals, and populations in the Liolaemus elongatus-kriegi complex (Squamata: Liolaemidae) in Andean Patagonian South America. AB - Recovery of evolutionary history and delimiting species boundaries in widely distributed, poorly known groups requires extensive geographic sampling, but sampling regimes are difficult to design a priori because evolutionary diversity is often "hidden" by inadequate taxonomy. Large data sets are needed, and these provide unique challenges for analysis when they span intra- and interspecific levels of divergence. However, protocols have been designed to combine methods of analysis for DNA sequences that exhibit both very shallow and relatively deeper divergences. In this study, we combined several tree-based phylogeny reconstruction methods with nested-clade analysis to extract maximum historical signal at various levels in the poorly known Liolaemus elongatus-kriegi lizard complex in temperate South America. We implemented a recently descrirbed tree based protocol for DNA sequences to test for species boundaries, and we propose modifications to accommodate large data sets and gene regions with heterogeneous substitution rates. Combining haplotype trees with nested-clade analyses allowed testing of species boundaries on the basis of a priori defined criteria. The results obtained suggest that the number of putative species in the L. elongatus kriegi complex could be doubled. We discuss these findings in the context of the advantages and limitations of a combined approach for retrieval of maximum historical information in large data sets and with reference to the yet formidable unresolved issues of sampling strategies. PMID- 12746149 TI - Intraorganismal homology, character construction, and the phylogeny of aetosaurian archosaurs (reptilia, diapsida). AB - Character construction, the methods by which characters and character states are produced from observations of variation, is a crucial but poorly understood step in phylogenetic analysis. Alternative approaches are used in practice, but there has been relatively little investigation of their theoretical bases and analytical consequences. We reviewed three published numerical analyses of the phylogenetic relationships within the Triassic Aetosauria. Combined data from these studies were used to explore the impact of alternative approaches to character construction. Some previous aetosaurian characters represent parallel variations in the morphology of osteoderms from different body regions, and their independence is questionable, leading us to propose more composite alternative constructions. Phylogenetic analyses revealed that inferred relationships within the Aetosauria are in general poorly resolved and weakly supported by the available data and are sensitive to alternative approaches to character construction. Thus, the results from this and previous studies should not, for the most part, be accepted as robust hypotheses of aetosaurian interrelationships. The treatment of systems of intraorganismal (e.g. serial, antimeric) homologues, such as osteoderms, in character construction is discussed. Applied to parallel variations in systems of intraorganismal homologues, previous advice on choosing among alternative character constructions and Hennig's auxiliary principle agree in favoring a more composite approach, in accordance with common practice. PMID- 12746150 TI - Clarifying aetosaur phylogeny requires more fossils, not more trees--reply to intraorganismal homology, character construction, and the phylogeny of aetosaurian archosaurs (reptilia, diapsida). PMID- 12746151 TI - Component coding, three-item coding, and consensus methods. PMID- 12746152 TI - Popper and systematics. PMID- 12746153 TI - Using a null model to recognize significant co-occurrence prior to identifying candidate areas of endemism. PMID- 12746155 TI - Determining "reliable detection limits" (RDLs) for environmental analyses. AB - It is generally accepted that the method detection limit or MDL (defined in 40 CFR 136, Appendix B) provides protection against false positives 99% of the time. This is correct, but only for the next single measurement performed after the MDL is determined. Subsequent measurements are not protected against false positives with the same degree of confidence, and there is no protection for false negatives. This paper provides a simple cost-effective approach for estimating the "reliable detection limit." Unlike the MDL, the statistic may be used for an indefinite number of future measurements and minimizes false negatives. PMID- 12746156 TI - Promoting GMP implementation: developing training materials for the international audience. AB - This paper outlines the development of a CD-ROM training package entitled: The WHO Basic Training Modules on GMP, intended to support the creation of training courses aimed particularly at government compliance officials who inspect pharmaceutical manufacturing facilities. The material was created over a three year period in collaboration with a team of external experts, WHO regional and local offices, and Drug Regulatory Authorities of participating countries. The nine training workshops and courses that contributed to the development and evaluation processes were attended by approximately 240 participants from 47 countries. To date over 5,800 copies of the CD-ROM have been distributed. PMID- 12746157 TI - Information resources on quality available on the internet. AB - The Internet continues to provide an excellent resource for information on quality assurance concepts, regulations, and practices. A search using just the word "quality" produced over 42 million hits. The combination of "quality" and "assurance" yielded over 2 million hits. Presented here is a sampling of 100 quality assurance sites organized alphabetically by site name, and accompanied by a brief description of the information available at the site. The choice of which sites to include was based on the author's experience and familiarity with the QA profession, and was aimed towards providing examples in active areas of QA including business and manufacturing, good practice regulations (i.e., GxPs), information quality, medical practice, software quality, higher education, and quality of research. The 100 sites provide access to a broad array of documents, services, forums, and opportunities to exchange ideas, and include links to major national regulatory and standard setting bodies around the world. PMID- 12746158 TI - Serum zinc and alkaline phosphatase values in pediatric bone marrow transplantation patients. AB - Zinc (Zn) plays an important role in the maintenance of immune functions, including cellular/humoral immunity, and in the prevention of oxidative injury. Therefore, the maintenance of a normal Zn status may be important in bone marrow transplantation (BMT) patients. Serum Zn levels were determined in 35 children during the BMT period. In addition, as Zn-related factors, serum Cu levels and alkaline phosphatase (AP) activity were also measured. There was a significant decrease in Zn and AP values during the immediate post-transplant period (lowest at day +7) when compared to pre-BMT levels (p <.01). The patients who developed hypozincemia were more likely to be transplanted for a diagnosis of malignant disorder and were younger, and adverse events appeared to occur more frequently. This preliminary study suggests that maintaining a normal Zn status may be important in BMT patients and that Zn deficiency may be a risk factor for adverse events. PMID- 12746159 TI - The prevalence of low selenium levels in newly diagnosed pediatric cancer patients. AB - Low selenium (Se) levels have been found in assoiciation with high incidences of various types of adult cancer. Much less is known about this issue among pediatric cancer patients. Forty-two pediatric patients with a variety of newly diagnosed malignancies were divided into two groups, 20 with localized disease (LD) and 22 with widespread disease (WSD). Analysis of serum collected before the commencement of treatment showed that half the patients had low Se serum levels, lower and more common in WSD than in LD. There was no significant difference in the prevalence of low albumin levels among patients with low Se levels, and most of the newly diagnosed children did not suffer from malnutrition. It was concluded that Se deficiency is common among newly diagnosed pediatric cancer patients, Se levels are lower in WSD than LD, and low Se levels are more prevalent in WSD patients than in LD patients. PMID- 12746160 TI - The changing pattern of bloodstream infection in pediatric oncology patients in the United Arab Emirates. AB - A surveillance study was performed on the pattern and antimicrobial susceptibilities of microorganisms isolated from blood cultures in the main pediatric oncology unit in the United Arab Emirates. The only previous data were published in 1995. During 1998-2000, the pattern of bloodstream infection changed. Enterobacter spp. and Enterococcus spp. emerged as frequent pathogens, while Pseudomonas aeruginosa became less common. There was a high incidence (65/198) of polymicrobial bacteremia not previously reported. This study highlights the need for continuous surveillance of the pattern of bloodstream infection in pediatric oncology patients so that changes may be detected and antimicrobial treatment modified appropriately. PMID- 12746161 TI - Sedation with midazolam and ketamine for invasive procedures in children with malignancies and hematological disorders: a prospective study with reference to the sympathomimetic properties of ketamine. AB - Different pharmacological agents have been used for sedation in children undergoing invasive procedures. The authors prospectively evaluated the efficacy, the occurrence of adverse side effects, and cardiovascular parameters in midazolam and ketamine sedation for invasive procedures in children with malignancies and hematological disorders. A total of 183 invasive procedures were performed on 63 children (mean age 9.2 +/- 5.2 years). Intravenous sedation consisted of 0.1 mg midazolam/kg and 1.0 mg ketamine/kg. Incremental dosages of ketamine (0.33 mg/kg) were given if necessary to maintain deep sedation. Systolic and diastolic blood pressure, heart rate, and oxygen saturation were recorded. All 183 invasive procedures were successfully completed with satisfactory sedation levels in 170 procedures (92.9%; 95% CI:88.2-96.2%). In 33 procedures (18%; 95% CI: 12.8-24.4%) sedation was associated with side effects, the most common being oxygen desaturation. One patient experienced a transient episode of laryngospasm. There was a significant increase in both systolic and diastolic blood pressure and heart rate after ketamine medication (p <.01). Procedure and recovery time were correlated to ketamine dosage (p <.01). The combination of midazolam and ketamine is efficacious in achieving deep sedation for painful invasive procedures. Considering the possibility of potentially serious respiratory complications it should be performed only by physicians who are trained in advanced airway management and life support. As opposed to many other sedative drugs with cardio-depressant properties, ketamine causes a rise in both systolic and diastolic blood pressure, and heart rate. PMID- 12746162 TI - Assessment of chosen parameters of the immune system in children with acute lymphoblastic leukemia. AB - The immunosuppressive effect of cytostatics, basic therapeutic agents in the treatment of proliferative diseases of the hematopoietic system, and the rising number of children cured from acute leukaemias form together a need to monitor the status of the immune system following cessation of therapy. Surface antigens in lymphocytes from peripheral blood were assessed in 16 children directly after intensive chemotherapy (i.e., after protocol II of the BMF program) and in 25 children 12-13 months following conclusion of acute lymphoblastic leukemia treatment. The results were compared to data obtained from children never afflicted with neoplastic disease. A significant decrease in the average number of lymphocyte subpopulations was noted in the case of the treated children directly after intensive chemotherapy. The average values of lymphocyte subpopulations in children with concluded treatment are within the norm, with the exception of NK and TS lymphocytes. PMID- 12746163 TI - Glomerular filtration rate in children with solid tumors: normative values and a new method for estimation. AB - Many chemotherapy regimens used in children are nephrotoxic. Accurate dosing of these medications requires that some estimation of glomerular filtration rate (GFR) be performed prior to initiating chemotherapy. However, few studies evaluating normal GFR in children exist. The authors report normal values for GFR for children with nonhematogenous malignancies using a highly accurate method of directly measuring GFR and an equation for estimating absolute GFR in these children. Children with nonhematogenous malignancies with no evidence of renal involvement or prior use of nephrotoxic agents had their GFR measured using an iothalamate infusion methodology. A total of 111 children (males and females) with a mean age = 7.95 years (range 2.8 months-19.5 years) were included in the study. GFR adjusted for body surface area (mL/min/1.73 m(2)) increases in the first 2 years of life and then plateaus at a level comparable to adult values. GFR adjusted for body surface area for males >2 years = 131.3 +/- 22.5, females = 126.8 +/- 24.4 mL/min/1.73 m(2) (p value not significant). Absolute GFR in mL/min can be easily estimated by a simple formula (r(2) =.97) based on the child's weight and serum creatinine: GFR (mL/min) = k sqrt[ ( (agemos+ 6)* wt) / Cr serum ] where agemos is age in months, wt is weight in kg, and k = 1.05 for males and 0.95 for females. The accurate measurement of GFR remains vitally important in the safe and effective treatment of pediatric solid tumors. This study provides a set of normal GFR values for these children and an equation for easy estimate of absolute GFR. PMID- 12746164 TI - Effect of twice weekly versus daily iron treatment in Turkish children with iron deficiency anemia. AB - This study was designed to propose a more practical, effective, safer, inexpensive, and manageable alternative treatment of iron deficiency anemia (IDA) for the developing countries. The study involves 94 children between the ages of 5 months and 6 years who had been seen in the authors' hospital and diagnosed as having iron deficiency anemia. Ninety-four children with IDA were randomly divided into two groups: 48 children comprised the first group, which was administered conventional treatment, and 46 children comprised the second group, which was administered intermittent treatment involving iron administration 2 days a week. Twenty-three children whose age and gender distribution were compatible with the other groups were included in the study as the control group. Both groups were reevaluated for their initial hematologic parameters at the end of the treatment. When the parameters of both groups were compared with the parameters of the control group after the treatment, there were no differences between hemoglobin, hematocrit, red blood cell, mean corpuscular volume, mean corpuscular hemoglobin concentration, serum iron, and ferritin levels of conventional and intermittent treatment groups. With respect to certain parameters, such as red cell distribution, serum iron binding capacity, transferrin saturation, transferrin receptor, and transferrin receptor/log ferritin, however, intermittent treatment was superior to the conventional treatment method (p <.05). In IDA, when a conventional treatment method or an intermittent treatment method is used, there are no differences between the hematological parameters. In fact, the intermittent treatment method has been found to be superior in many parameters. PMID- 12746165 TI - Cerebral atypical teratoid/rhabdoid tumor of infancy: long-term survival after multimodal treatment, also including triple intrathecal chemotherapy and gamma knife radiosurgery--case report. AB - Cerebral atypical teratoid/rhabdoid tumors (AT/RT) of infancy are highly malignant and have a poor prognosis. The authors report on one case with long term survival. The patient was a 1 year-old boy presenting with a large AT/RT in the right temporal lobe. He was treated with complete surgery, followed by multiagent chemotherapy. Later he had a second resection and intrathecal chemotherapy and Gamma knife radiosurgery was added to the treatment. Except for a well-controlled temporal epilepsy, the boy is doing well after 6 years follow up. AT/RT should be treated in a multimodal way. Intrathecal chemotherapy and Gamma knife radiosurgery of single recurrent or residual tumors might increase survival. PMID- 12746166 TI - Primary epidural burkitt lymphoma in a child: case presentation and literature review. AB - Primary spinal epidural Burkitt lymphoma, presenting with signs of spinal cord compression, is very uncommon in childhood. Previously reported pediatric cases with isolated epidural Burkitt lymphoma had a high mortality, and survivors usually suffered serious neurologic sequelae. The authors present a 13-year-old female with isolated epidural Burkitt lymphoma with favorable outcome, and review the pediatric literature. PMID- 12746168 TI - High-dose methylprednisolone treatment of hepatic veno-occlusive disease in a child with Wilms tumor. AB - Veno-occlusive disease (VOD) is a rare complication of chemotherapy in children with cancer. In the literature successful treatment of severe VOD has not been well determined. The authors report a 5-year-old boy with Willms tumor whose condition was complicated with VOD while receiving chemotherapy. He was treated successfully with high-dose methylprednisolone. After therapy, the clinical signs of VOD, such as abdominal ascites, oliguria, and jaundice, disappeared without any adverse effects of therapy. PMID- 12746167 TI - Anti-CD20 monoclonal antibody (Rituximab) for therapy of mediastinal CD20 positive large B-cell non-Hodgkin lymphoma with a local tumor extension into the lung of a 10-year-old girl. AB - Intravenous therapy with the anti-CD20 antibody Rituximab has been recently approved for the treatment of CD20 positive non-Hodgkin's lymphoma (NHL) in adults but not in children. The authors present the benefits of its application for mediastinal NHL CD 20+ with a local extension into the lung of a 10-year-old girl. Receiving the chemotherapy according to study NHL-BFM-95 for high-risk lymphoma the girl did not reach complete remission. Before the last chemotherapy block was started, a computed tomography scan of the thorax showed residue in the right lung. Twenty-five days after the last chemotherapy she received Rituximab at a dose of 375 mg/m(2) by intravenous infusion once a week for a total of four doses without the adverse reactions. Complete remission was achieved. The patient was high risk with lung involvement of lymphoma suggesting a pure prognosis. The results suggest that Rituximab may improve the outcome in high-risk patients and appeared to be safe and effective in children also. PMID- 12746169 TI - Giant-cell tumor of the rib in a 12-year-old girl: a case report. PMID- 12746171 TI - Case report: congenital salivary gland analage tumor presenting with neonatal respiratory distress. AB - Stridor in the newborn is not an unusual entity. This article describes a rare presentation of neonatal respiratory distress caused by a benign nasopharyngeal salivary gland analage tumor. PMID- 12746170 TI - The role of fibroblast transdifferentiation in lung epithelial cell proliferation, differentiation, and repair in vitro. AB - Parathyroid hormone-related protein (PTHrP) expression is necessary for differentiation of mesenchymal lipofibroblasts, which induce epithelial type II (TII) cell differentiation, both of which are necessary for alveolarization. PTHrP deficiency may be associated with bronchopulmonary dysplasia (BPD), characterized by truncation of alveolarization among preterm infants. This is supported by the baboon model of BPD (failure of alveolarization) that manifests PTHrP deficiency. We provide evidence that TII cell PTHrP expression is downregulated by alveolar overdistension, resulting in the transdifferentiation of lipofibroblasts to myofibroblasts, characterized by progressive loss of PTHrP receptor expression and triglyceride content, and sequential upregulation of alpha-smooth muscle actin (alphaSMA), typifying fibrosis. PTHrP reverses the downregulation of the PTHrP receptor and upregulation of alphaSMA, reverting myofibroblasts to a lipofibroblast genotype. When TII cells are co-cultured with lipofibroblasts, they proliferate and differentiate, expressing surfactant protein-B; in contrast, TII cells co-cultured with myofibroblasts fail to develop, mimicking the failed alveolarization associated with BPD. Treatment of myofibroblasts with 15-deoxy-Delta 12, 14 prostaglandinJ(2) (PGJ(2)) stimulates ADRP expression, reconstituting the lipofibroblast phenotype. PGJ(2)-treated myofibroblasts promote TII cell growth and surfactant protein-B expression, indicating that failed alveolarization due to transdifferentiation is reversible. We conclude that alveolar overdistension can cause fibroblast transdifferentiation, resulting in failed alveolarization. PMID- 12746172 TI - Detection of n-myc gene amplification in neuroblastoma by comparative, in situ,and real-time polymerase chain reaction. AB - We have used semiquantitative comparative and real-time quantitative polymerase chain reactions (PCR) to detect n-myc gene-amplification in 20 frozen neuroblastoma biopsies and IMR 32 cell line to predict biological behavior of the tumors. Two primer pairs were used for the semiquantitative method to co-amplify a 520-bp fragment of the beta-globin gene--used as a single copy reference standard--and a 258-bp fragment of the n-myc gene. After 30 cycles the PCR products were electrophoresed through an agarose gel and were compared to each other with use of a gel-densitometer. Real-time quantitative analyses were performed in a LightCycler instrument. A single primer pair was used to amplify a 120-bp fragment of the n-myc oncogene and a LC640-labeled fluorescent probe pair to detect the product. Calibration curve, set up from a serial dilution including samples with 1, 2, 10, 13, 25-fold n-myc oncogene amplification, was used for quantitative analysis. The semiquantitative method did not show distinct difference between tumor groups with no amplification and less than 10-fold amplification, whereas quantitative LightCycler analysis was able to detect even 2-fold amplification. Differentiated neuroblastomas seldom show n-myc amplification. In spite of this, we have found two partly differentiated tumor samples that contained n-myc amplification. In these cases in situ PCRs were performed to examine the tumor heterogeneity. We used biotinated ATP labeling and the same primer pair as for the LightCycler analysis. In both cases differentiated cells did not show n-myc gene amplification, whereas considerable amplification was detected in the neuroblasts. PMID- 12746174 TI - Familial hemophagocytic lymphohistiocytosis: an autopsy study. AB - We describe four classical cases of familial hemophagocytic lymphohistiocytosis (FHL), a macrophage-related, autosomal recessive fatal disorder. Parental consanguinity was present in three cases. All patients presented with fever, neurological involvement of varying degrees, hepatosplenomegaly, cytopenias, deranged liver function tests, and coagulogram, hypofibrinogenemia (three cases), and hyperlipidemia (one case). An antemortem diagnosis could not be made, although it was suspected in one case. Necropsy (done in three cases and postmortem liver biopsy in one case) revealed classical features of FHL. Florid lymphohistiocytic infiltrate exhibiting hemophagocytosis was seen in the bone marrow, liver, spleen, lymph nodes and brain (examined in two case). In addition to this, focal infiltrates were seen in the kidneys, lung, pancreas, testes, adrenals, and skin. Marked lymphoid depletion was seen in one case in the lymph nodes and spleen. PMID- 12746173 TI - Placental calcium pump: clinical-based evidence. AB - Placenta can be considered as a pump of calcium necessary for the normal development of the fetus. We believe that the location of this pump is in the placental basement membrane. The calcification of this membrane has been described only in cases of in utero fetal death. In this study we describe for the first time a case of placental calcification in a living fetus. The fetus of a normal 21-year-old pregnant woman showed heart abnormalities but the genetic analysis showed a normal male karyotype. The histology of the placenta demonstrated multiple intravillous linear and granular calcific incrustations The hemtoxylin/eosin stain of the sections revealed basement membrane calcific incrustations and intravillous calcium deposits. We postulate that the fetal circulation in the villi was impaired and the calcium that reached the villi from the mother was deposited at this level. PMID- 12746175 TI - Pathology of the umbilical cord in adrenal fusion syndrome. AB - We report the placental findings of a newborn with multiple anomalies that include small and fused adrenal glands, meningomyelocele, lobar holoprosencephaly, cerebellar hypoplasia, short pancreas, and small spleen, fitting with the features of the so-called adrenal fusion syndrome. The placenta was small and presented a short, undercoiled and segmentally uncoiled umbilical cord with a short constriction in the middle of its length. We speculate that adrenal fusion syndrome and abnormal spleen lobulation and short pancreas may represent part of the same complex and probably related to the one including midline anomalies and defects of laterality formation in which horseshoe (fused) adrenal gland also may be found. Undercoiled umbilical cord might represent the placental component of this malformation complex. We also underline the value in identifying abnormalities of the umbilical cord to establish the cause of death. PMID- 12746177 TI - Pathology teach and tell: pediatric gastrointestinal stromal tumor. PMID- 12746176 TI - Reproductive toxicity of the psoralens. AB - The psoralens are naturally occurring secondary metabolites in plants, including many fruits and vegetables. Health risks have been associated with handling or ingesting psoralen-containing plants, and with the use of synthetic psoralens in photochemotherapy of skin disorders. Our research has demonstrated that administration of the psoralens bergapten (5-methoxypsoralen) and xanthotoxin (8 methoxypsoralen) in the diet of female rats reduced birthrates, number of implantation sites, pups, corpora lutea, full and empty uterine weight, and circulating estrogen levels in a dose-dependent manner. Psoralens induced mRNAs of the liver enzymes CYP1A1 and UGT1A6, suggesting that enhanced metabolism of estrogens by psoralen treatment may explain the reproductive toxicity and the observed reduction of ovarian follicular function and ovulation. Rats also avoided repeated consumption of a flavored solution associated with psoralen administration. The findings indicate that the psoralens constitute a novel group of ovarian toxicants. Further examination of the safety of their use in photochemotherapy and diet is warranted. PMID- 12746178 TI - Pathology teach and tell: congenital myelodysplastic syndrome with extensive hepatic fibrosis. PMID- 12746179 TI - From the editor--toward increased global dialogue about violence reduction. PMID- 12746180 TI - Consumers/service users: is nursing listening? AB - This article synthesizes some consumer literature and highlights the fact that many of their priorities fall outside the purview of contemporary mainstream psychiatry. Given that there is insufficient integration of consumer information into mental health nursing literature, the author overviews biopsychiatry's emphases and omissions for clues to the marginalization of consumer-based concerns. As nursing traditionally focuses on activities of daily living impeded by illness or disability (unlike psychiatry), the article argues that we have a responsibility to assist consumers manage life difficulties arising from structured constraints such as unemployment, poverty, insecure accommodation, and stigma. PMID- 12746181 TI - Stress discourse and Western biomedical ideology: rewriting stress. AB - Popular psychosocial theories of stress and coping are based on an empiricist paradigm and a Western biomedical framework that reflect Western ideologies and values about health and illness. Problems associated with this discourse lie mainly in the ideologies that emphasize naturalism, individualism, rationalism, and objectivity. We suggest that stress and coping theory should be concerned with the ways in which power relations and social institutions produce the meaning of stress. The inclusion of alternative discourses that attend to the micro and macro social and historical factors is necessary for the further development of stress and coping theory and practice. PMID- 12746182 TI - Assaults and nonpatient-induced injuries among psychiatric nursing staff in Bahrain. AB - This retrospective study of incident reports at the psychiatric hospital in Bahrain was carried out to describe nursing staff injuries that were reported during 1992-1999. The average assault rate (4.4%) was much lower than that reported in Western countries. Of the 111 injuries, 44.0% occurred in wards assigned for patients with acute conditions, 27.5% in chronic condition wards, 5.5% in outpatients, and 22.9% in the community. Staff assaults by patients constituted 60.4% of all injuries, of which 64.2% occurred in the 1992-1995 period. The assault rate was highest in 1992 (6.8/100) and lowest in 1998 (1.1/100), while the rate for all injuries was highest in 1994 (12.0/100) and lowest in 1998 (3.3/100). Bahraini staff had higher assault (7.0/100) and total injury (14.8/100) rates than the non-Bahraini (5.5, 6.8/100, respectively). Odds ratios for assaults vs. nonpatient-induced injuries indicated that assaults were 2.3 times (95% CI: 1.05-4.95) more likely to occur in males, 5 times more likely (95% CI: 1.99-12.15) in non-Bahraini, 1.79 times more likely (95% CI: 0.81-3.95) in staff nurses, and 2.3 times more likely (95% CI: 1.05-5.01) in the evening. Suggestions for reducing assaults and nonpatient-induced injuries are discussed. PMID- 12746184 TI - Shifting physical health care responsibilities at a community mental health center. AB - This study assesses the effects of transferring physical health care of consumers from non-nurse case managers to a nurse case manager at a community mental health center. Using a comparative descriptive design, pre- and postintervention surveys were distributed to clinical staff before and after the transfer of responsibilities to determine differences in responses relating to workload and quality of consumer care. Findings suggested that staff had more time to spend on treatment consistent with their education and training, and experienced improved job satisfaction. They reported that consumers' health care improved in terms of quality, efficiency, access, continuity, and follow-up. A chart review revealed that the number of current annual health histories decreased slightly (6%), but annual physical exams increased by 24%. Types of medical appointments were analyzed to note the complexity of health needs of the consumers, with 38% being for routine care and the remaining 62% for chronic, specialty, and acute care. Nurse case managers responsible for overseeing consumers' physical health care would be a valuable addition to community mental health centers. This study suggests improved consumer care and job satisfaction ramifications. PMID- 12746183 TI - Women's conceptions of coping with major depression in daily life: a qualitative, salutogenic approach. AB - The experience of having a severe disease such as major depression affects all aspects of the individual's life, including family, work, and social functioning. Therefore, the aim of this study was to describe, from a salutogenic approach, women's conceptions of coping with major depression in daily life with the help of professional and lay support. Thirteen women, previously hospitalized for major depression, were included in the study. The women were selected by strategic sampling, and data were analyzed by application of a phenomenographic approach. Four descriptive categories emerged: Self-Healing, Managing, Receiving Social Support, and Finding Meaning. While working their way out of the depression, the women needed to undergo a process of transition, involving both a cognitive and an emotional understanding, which they subsequently translated into health-related actions. The task of psychiatric mental health nurses is to provide care that empowers patients. Future nursing research should explore the circumstances that empower these women to start the transition process, as this process appears to be vital for recovery from major depression. PMID- 12746186 TI - The art in surgery: the relevance of artistic expression in the surgeon's life. PMID- 12746187 TI - William Beaumont: first American surgeon scientist and father of gastric physiology. PMID- 12746188 TI - Upregulation of endogenous heparin-binding EGF-like growth factor (HB-EGF) expression after intestinal ischemia/reperfusion injury. AB - Expression of endogenous heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF), a proven intestinal cytoprotective molecule, was examined in intestinal epithelial cells (IEC) in vitro, and in intestine undergoing ischemia/reperfusion (I/R) injury in vivo. In vitro, cells were exposed to anoxia for 90 min followed by reoxygenation for 1-3 h (A/R). In vivo, total midgut I/R injury was produced in rats by occlusion of the superior mesenteric artery for 30 or 90 min followed by reperfusion for 4 h. In situ hybridization and immunohistochemistry were used to study HB-EGF mRNA expression and protein production. In vitro, normal IEC had no detectable HB-EGF mRNA or protein expression. After anoxia, cells expressed HB-EGF mRNA and protein, with expression reaching a peak 2-3 h after reoxygenation. In vivo, only very low levels of HB-EGF mRNA and no detectable protein were found in normal intestine. Four hours after I/R, HB-EGF protein was detected in villous epithelia subjected to 30 min but not 90 min of ischemia, whereas HB-EGF mRNA was highly expressed after both ischemic intervals. Endogenous HB-EGF is immediately upregulated in IEC after A/R injury and in intestine after I/R injury. Thus, HB-EGF acts as an immediate early gene under these conditions. PMID- 12746189 TI - Peritoneal release of TNFalpha and IL-6 after elective colorectal surgery and anastomotic leakage. AB - The evaluation of postoperative peritoneal drainage fluid tumor necrosis factor (TNF)alpha and interleukin (IL)-6 was studied prospectively over a 7-day period in 25 patients operated on for neoplastic colorectal diseases. In 22 cases, colon or rectum carcinoma was the reason for surgery, and in 3 patients resection was performed because of colonic adenoma. All patients received either an end-to-end colo-colonic or colorectal anastomosis. Of this group, 22 patients were free of complications defined as uneventful postoperative course without any signs of anastomotic leakage until the 14th postoperative day. All of these patients showed a significant rise in peritoneal TNFalpha with maximum on the 7th day during the study period (p <.05). In contrast, peritoneal IL-6 levels remained constant without significant change in time (p >.05). Three patients underwent relaparotomy because of anastomotic leakage. In these patients, peritoneal TNFalpha concentrations showed a rise until the day of operative confirmation of anastomotic leakage. This rise preceded the day of operative confirmation by at least 1 day but did not change significantly in time (p =.59). Peritoneal IL-6 concentrations in patients with anastomotic leakage remained constant and also did not change significantly in time (p =.21). After elective colorectal surgery, neither postoperative abdominal drainage fluid TNFalpha nor IL-6 monitoring is helpful to decide on the need for revision in patients with anastomotic leakage. PMID- 12746191 TI - Recovery of preservation-injured primary human hepatocytes and nonparenchymal cells to tissuelike structures in large-scale bioreactors for liver support: an initial transmission electron microscopy study. AB - This study investigated large-scale regeneration and tissue reorganization of adult human liver cells from preservation injured transplant organs. The use of basement membrane protein gels and growth factor enriched culture medium in standard culture flasks promotes liver tissue formation in isolated rat and pig hepatocytes, resulting in prolongation of phenotypic stability and metabolic competence of primary cells in vitro. A special bioreactor construction for high density three-dimensional cell recovery was developed and isolation of cells from discarded human donor livers was enabled. In vitro regeneration of adult human liver cells isolated from preservation-injured organs took place over a period of 2 weeks in a purpose-built bioreactor. Basement membrane protein and growth factors were avoided. Reorganization of tissue structures was studied using transmission electron microscopy (TEM). This showed regeneration and tissue reorganization of adult human cells from preservation-injured organs by coculture with nonparenchymal cells in the bioreactor. The majority of the aggregated hepatocytes in the bioreactors showed morphological similarities to those in vivo (although not re-formed to hepatocyte plates), exhibiting cell-cell junctions and reconstituted bile canaliculi-like spaces between neighboring hepatocytes. Perfusion channels appeared regularly between cell aggregates. The arrangement of nonparenchymal cells between the hepatocyte aggregates exhibited similarities to liver sinusoids. Endothelial cells often covered the aggregates and formed a borderline to the perfusion channels between the capillaries. Similar to the space of Disse, further nonparenchymal cells were located between the endothelial cells and the parenchymal aggregates. Deposits of biomatrix fibers occurred spontaneously. The regenerated cell mass was close to that of a single liver lobe. In conclusion, the further optimization of bioreactors that enable cell recovery from preservation injury may lead to the utilization of cells from discarded whole or split transplants for extracorporeal temporary liver support therapy or hepatocyte transplantation. PMID- 12746190 TI - Piroxicam-induced regression of intestinal adenomatous polyps in APC(delta474) mice. AB - Mutation of adenomatous polyposis coli (APC) gene results in incidence or development of polyps and colorectal cancer. It has been reported that nonsteroidal anti-inflammatory drugs (NSAIDs) inhibit cell growth, cause cell cycle arrest, and induce apoptosis. The aims of this study are to investigate chemopreventive effects of piroxicam and elucidate its mechanism. All APC(delta474) mice have intestinal polyps. Thirty-five APC(delta474) mice were divided into three groups: 0.005% solution of piroxicam in tap water was given for P group (n = 15) and 0.001% solution for P' group (n = 5), and water without piroxicam for C group (n = 15) from 4 weeks of age to 12 weeks, respectively. All mice were sacrificed at the 12th week after birth. Hematoxylin-eosin staining for number and size of polyps, immunohistochemical staining for cyclooxygenase (COX) 1 and -2, proliferating cell nuclear antigen (PCNA), vascular endothelial growth factor (VEGF), TUNEL method, and Western blot analysis of COX-2 and VEGF were performed. Polyps were divided into two types of large polyps of >or=300 microm in diameter and small polyps of <300 microm. The number of large polyps in P group decreased significantly compared with C group (p <.0001), but without significant difference in small polyps. There were no significant differences in PCNA index in both of large and small polyps among the three groups. Apoptotic index of polyps in P group increased more than those in C group (p <.05). There was immunohistochemically no significant difference in COX-1 positivity of normal intestinal epithelia and adenomas among three groups. Both numbers of VEGF positive cells and COX-2 positive cells in the stroma of the small intestine were significantly downregulated in P group (p <.05). COX-2 expression was inhibited in dose-dependent manner without significant difference. There were no significant differences in VEGF expression between P' and C groups. In conclusion, piroxicam suppressed the development of large polyps in APC(delta474) mice by inducing apoptosis and inhibiting VEGF expression in interstitial cells of polyps. PMID- 12746192 TI - Strangulated ventral hernia model in rats. AB - This study was performed to describe a new model of strangulation obstruction mimicking the situations relevant to abdominal hernias. The strangulation obstruction was induced either by intra-abdominal ligation of an ileal segment 2 cm in length (n = 20) or by a pursestring suture around a fascial defect with a strangulated intestinal loop placed subcutaneously (n = 20). Ten animals served as sham operated controls (n = 10). All animals were euthanized at 12 h postoperatively; strangulated ileum segments were collected for histopathological examination. Microscopic injury scores were similar in both strangulation groups, which were significantly different from the control group (p <.001). The model described here seems to be appropriate for use in further experimental studies concerning strangulation obstruction injury and its consequences, with the added advantage of visualization of the strangulated intestinal loop beneath the skin. PMID- 12746196 TI - UltraPath XI: celebrating twenty years. PMID- 12746193 TI - Partial hepatectomy in the mouse: technique and perioperative management. AB - Hepatic surgery in mice is challenging because of the delicate nature of the liver, lack of intravenous access, and risk of hemorrhage. In order to study the ability of the liver to regenerate after surgical resection, we developed a novel, rapid, and safe technique for partial hepatectomy in mice. We determined the relative contributions of the seven lobes of the mouse liver and resected the three most anterior lobes for a 68% hepatectomy. We used general anesthesia, a small upper midline incision, silk suture to tie off the lobes to be resected, warming pads and lights, as well as subcutaneous saline injection to ensure minimal morbidity. We have performed a safe two-thirds hepatic resection in 288 of 300 C57BL6 mice (96%). Perioperative mortality was due to technical error. Minimal long-term morbidity was appreciated. This technique may be applied to any type of hepatic resection in mice. In addition, the general operative technique and perioperative management of these mice may be applied to all types of murine intra-abdominal procedures used for surgical research. PMID- 12746197 TI - Intestitial cells of Cajal in the human small intestine: immunochemical and ultrastructural study. AB - The stem cell kinase CD117 has recently been found to play an important role in the development of interstitial cells of Cajal (ICC), which are currently regarded as pacemaker cells of the gastrointestinal tract. CD117 is expressed in both gastrointestinal stromal tumors (GIST) and ICC, with the latter regarded by many as the progenitor cells of GIST. The authors investigated immunoreactivity of 25 normal surgically removed small intestinal tissues and correlated the findings with electron microscopy (EM) on 12 cases. In all cases CD117-positive cells were frequently seen around the myenteric plexi either singly or in groups. CD117-positive cells on immunostained sections corresponded to the cells appearing as fibroblast-like or undifferentiated primitive mesenchymal cells around the myenteric ganglia and interstitial spaces by EM. In contrast, S-100 stain revealed a fine network of positive staining throughout the muscularis. Branches of nonmyelinated axons and nerve endings were found regularly between myocytes with direct contact with muscle cells by EM. The cells that we could depict as ICC because of their distribution and staining pattern of CD117 were limited to the nonmuscular mesenchymal cells. No muscle cell-like ICC were found. Instead, the muscle cells in direct contact with nerve endings were often disfigured and the cytoarchitectural contents for muscle cells became less distinct because of lighter staining and loss of definite focal densities among actin filaments. However, these latter cells did maintain most muscle cell features, such as continuous external lamina, caveolae, and some of the peripheral densities. These findings raise a possibility that previous investigators could have included these altered muscle cells into the ICC group. It was also found that intestinal muscularis not only was richly endowed with an elaborate neural network of delicate axonal extensions and dense-core granule containing nerve endings traversing through and between myocytes, but also showed frequent synapse-like direct contact between nerve endings and muscle cells. These findings indicate that enteric nerves may play a major role in the control of intestinal motility, while CD117-positive cells play an accessory role as cells of Cajal as originally speculated. Further studies are necessary to better define and characterize interstitial cells of Cajal, which will be useful in the correlation of the vast number of data concerning the possible role of CD117 positive ICC in the pacemaker function of the intestine and oncogenesis of GIST. PMID- 12746198 TI - Tactile corpuscle-like bodies in colonic mucosa. AB - During the routine examination of a segment of colon resected for adenocarcinoma, a diffuse proliferation of mucosal tactile corpuscle-like bodies was identified. The bodies showed a lamellar structure by light microscopy and were S-100 positive. Electron microscopy demonstrated parallel slender processes with prominent surface caveolae, arising from peripheral cell bodies. Similar structures sometimes occur in neurofibromas but they have not previously been reported in the gastrointestinal tract. PMID- 12746199 TI - Parietal cell carcinoma of gastric cardia: immunophenotype and ultrastructure. AB - The case is reported of a clinically aggressive parietal cell carcinoma of the gastric cardia in a 67-year-old man. Histologically, the tumor was a poorly differentiated adenocarcinoma with a predominantly solid growth pattern, though with areas exhibiting glandular morphology and with extensive lymphatic invasion. The tumor cells had eosinophilic, finely granular cytoplasm, with focal Alcian blue-positive mucin in the gland lumens. Ultrastructural examination of the pleural metastasis and gastrectomy specimen demonstrated many mitochondria, tubulovesicular profiles of endoplasmic reticulum, and intracytoplasmic lumens, which resembled intracellular canaliculi of parietal cells. Immunohistochemically, there was positive staining of tumor cells for the parietal cell specific antibodies to H/K-ATPase and human milk fat globule-2 (HMFG-2). PMID- 12746200 TI - Metastatic epithelioid gastrointestinal stromal tumor: yet another tumor with anemone cell features. AB - Since 1980 when Sibley and coworkers first described a nodal neoplasm of unknown histogenesis with striking surface microvilli for which they introduced the term "anemone cell," a series of reports have appeared in the literature illustrating tumors with similar ultrastructural features. While most reported cases showed differentiation along a particular line, rare cases remained histogenetically unclear. In this report a case is described of epithelioid gastric gastrointestinal stromal tumor metastatic to the liver, showing conspicuous long microvillus-type cell processes partially or circumferentially coating the cell surfaces, thus qualifying as yet another tumor type with anemone cell features. PMID- 12746201 TI - Ultrastructural spectrum of medulloblastoma with immunocytochemical correlations. AB - Electron microscopy was used to examine 72 cases of medulloblastoma to better characterize the ultrastructural spectrum of this tumor. Twenty-four cases showed prominent neural differentiation. Twenty-three cases showed minimal (21) or no (2) recognizable neural differentiation, and the remainder of the cases (25) showed intermediate differentiation. All 42 cases tested stained for neuron specific enolase, 28 for synaptophysin, and 12 for neurofilament protein. All cases showed strong reactivity for glial fibrillary acidic protein (GFAP) within reactive astrocytes. Three cases showed reactivity for GFAP within tumor cells. Medulloblastoma exhibits a broad spectrum of neural differentiation, with nearly all cases showing at least some degree of this change, and it universally exhibits participation of reactive astrocytes which can create a potential for diagnostic confusion. PMID- 12746203 TI - Pneumocystis carinii: an update. AB - Pneumocystis produces respiratory infection in immunocompromised individuals of several species of mammals, including humans. Each mammalian species has its own specific Pneumocystis species, which does not cross-infect other mammals. The species infecting humans has now been renamed P. jerovici, since P. carinii is reserved for one of two species infecting rats. Long believed to be a protozoan, Pneumocystis is now classified as an Archiascomycetous fungus. This is based on new molecular taxonomic techniques using DNA sequence analysis of srRNA genes. Only two of about 140 copies of the gene that exist in Pneumocystis were used for sequencing, so the evidence is not conclusive; however, it is supported by morphological evidence such as fungus-specific nucleus-associated organelles for cell division. There is also ultrastructural evidence of meiotic division and sexual conjugation. Clinically, several lines of evidence suggest the improbability of latent infection. Adult infections appear to be new infections, a fact that invites a new perspective on prevention. PMID- 12746202 TI - Cerebellar central liponeurocytoma. AB - Cerebellar liponeurocytoma is a rare, benign neuroepithelial tumor that occurs exclusively in the cerebellum of adults. Its salient histological features include advanced neuronal/neurocytic differentiation, focal vacuolated cells resembling mature adipose cells, low mitotic activity, and lack of endothelial proliferation and/or necrosis. The morphological appearance of this neoplasm can be confused with that of oligodendroglioma, neurocytoma, ependymoma, medulloblastoma, hemangioblastoma, metastatic renal cell carcinoma, and other clear cell carcinomas. Its full biological potential and histological features, however, have not been fully exploited due to the rarity of this tumor. The authors describe a case with clinical, imaging, histological, immunohistochemical, and ultrastructural features. PMID- 12746204 TI - Absence of nexin links as a possible cause of primary ciliary dyskinesia. AB - Transmission electron microscopy of nasal cilia was performed in three patients, two of them siblings, with repeated respiratory infections. Number of microtubuli and dynein arms were within normal limits and they had an ordered arrangement except for a disarray of the microtubuli in some areas of the biopsies from two of the patients. In the normal areas radial spokes and sheaths were easily found but nexin links could not be discerned in any of the patients. The orientation of the cilia was partly random. As all patients repeatedly and constantly had very low nasal NO (range 9-15 ppb; normal findings for persons <10 years old are > 50 ppb), the diagnoses were very likely primary ciliary dyskinesia (PCD). Absence of nexin links may be an ultrastructural variant of PCD. Deficiency of these structures might be the cause of the microtubular disarray observed in some areas of the biopsies. PMID- 12746205 TI - Titanium particles identified by energy-dispersive X-ray microanalysis within the lungs of a painter at autopsy. AB - A 72-year-old male painter, who complained of his "lungs burning" for 2 weeks, died suddenly. Autopsy examination revealed severe coronary atherosclerosis with plaque rupture as the cause of death. Examination of the lungs revealed emphysema, interstitial fibrosis, and multinucleated giant cells with intra- and extracellular brown-black, crystalline, polarizable foreign material. Energy dispersive X-ray microanalysis showed the material to contain titanium, aluminum, silicon, and iron. An increased incidence of respiratory disease has been reported in professional painters. Titanium is widely used as a pigment in the manufacturing of commercial paints. Cases of pneumoconiosis and alveolar proteinosis have been described in painters in which analysis of lung tissue revealed increased levels of titanium. This case is presented as an example of a rarely reported phenomenon, which may have clinical implications for evaluation and management of lung disease in painters. PMID- 12746206 TI - Success factors for open access. AB - Open access to the peer-reviewed primary research literature would greatly facilitate knowledge transfer between the creators and the users of the results of research and scholarship. Criteria are needed to assess the impact of recent initiatives, such as the Budapest Open Access Initiative. For example, how many open-access research journals exist within a given field, and what is the reputation of each one? And, how many openly-accessible institutional e-print archives have been created and how many are actually are being used by researchers and scholars? A simple approach to an assessment of the open-access portion of the medical literature is described, and some preliminary results are summarized. These preliminary results point to the need for incentives to foster the implementation of initiatives such as the Budapest Open Access Initiative. An example of an incentive model is proposed, where an agency or foundation that provides peer-reviewed grants-in-aid to researchers establishes an e-print archive. Only current grantees of the agency would be eligible to post reports about the results of research projects or programs that have been supported by the agency. Some advantages and implications of this particular model are outlined. It is suggested that incentive models of this kind are needed to increase the likelihood that open access to the primary medical research literature will soon reach a "tipping point" and move quickly toward wide acceptance. PMID- 12746208 TI - A model for online interactive remote education for medical physics using the Internet. AB - BACKGROUND: Medical physics is a relatively small community but it spans great geographical distances, usually with a scarcity of experts whose expertise could greatly benefit students entering into the field. In addition there are many software systems for which an interactive education method would be most advantageous. OBJECTIVE: To develop a process to optimally use the Internet for real-time interactive remote education of medical physics and to present the experience of the study. METHODS: The project is a collaboration of the Department of Medical Physics at the Toronto-Sunnybrook Regional Cancer Centre in Canada and the Department of Radiology at the University of Malaya in Malaysia. A class of medical-physics graduate students at the University of Malaya attended lectures provided by lecturers in Toronto, using the Internet as the main tool of communication. RESULTS: The different methods that can be used to provide the real-time interactive remote education were explored, and various topics including traditional classroom lectures as well as hands-on workshops-were also delivered. CONCLUSIONS: The concept of real-time interactive remote education is viable and holds promise for providing economical and practical tele-education to the medical physics community, but depends heavily on the availability of the Internet in many developing countries. PMID- 12746209 TI - eHealth in Latin America and the Caribbean: development and policy issues. AB - This paper reviews trends and issues in health and in the information and communication technologies (ICT) market as they relate to the deployment of eHealth solutions in Latin America and the Caribbean. Heretofore designed for industrialized countries and large organizations, eHealth solutions are being proposed as an answer to a variety of health-system management problems and health care demands faced by all health organizations including those in developing societies. Particularly, eHealth is seen as especially useful in the operational support of the new health care models being implemented in many countries. The authors examine those developments vis-a-vis the characteristics of the Latin American and the Caribbean health-sector organizational preparedness and technological infrastructure, and propose policy and organizational actions to foster the development of eHealth solutions in the region. PMID- 12746210 TI - Developing countries must invest in access to information for health improvements. PMID- 12746207 TI - LASIK complications and the Internet: is the public being misled? AB - BACKGROUND: LASIK (Laser in Situ Keratomileusis) is a very popular combined surgical and laser procedure, which is used to correct myopia (shortsightedness) and hyperopia (farsightedness). There is concern that the public is being misled regarding the safety of the procedure. OBJECTIVES: To assess the quality and quantity of the information on complications on LASIK Web sites. METHOD: Serial analysis and evaluation of the authorship, content, and technical quality of the information on the complications of LASIK on 21 Web sites. RESULTS: Of the 21 LASIK Web sites visited, 17 were commercial. Of the 21 Web sites, 5 (24%) had no information on complications. Of the 16 sites that had information on complications the author of the information was clearly identified in 5 (31%), the content was only referenced in 2 (12.5%), and evidence of the information having been updated was only seen in 2 (12.5%). The quantity of information is generally minimal and the information itself is generally difficult to understand and locate. CONCLUSION: The quality and quantity of the information on the Web on the complications of LASIK are poor. More work is required to encourage clear, accurate, up-to-date, clearly authored, and well-referenced, balanced ophthalmic information. PMID- 12746211 TI - Use of the Internet in scanning the horizon for new and emerging health technologies: a survey of agencies involved in horizon scanning. AB - BACKGROUND: A number of countries worldwide have structured horizon scanning systems which provide timely information on the impact of new health technologies to decision makers in health care. In general, the agencies that are responsible for horizon scanning have limited resources in terms of budget and staff. In contrast, the number of new and emerging health technologies, i.e. pharmaceuticals, medical devices, and medical and surgical procedures, is growing rapidly. This requires the Horizon Scanning Systems (HSSs) to devise efficient procedures for identification of new health technologies. The role of the Internet for this purpose has as yet not been documented. OBJECTIVE: To describe and analyse how the Internet is used by horizon scanning systems to systematically identify new health technologies. METHODS: A questionnaire was developed and distributed among 10 agencies known to work within this specific area. The questionnaire specifically focussed on type of sites scanned, frequency of scanning, and importance of a site for the identification of a new health technology. RESULTS: A 100% response rate was obtained. Seven out of 10 agencies used the Internet to systematically identify new health technologies, of which 6 provided complete information. A total of 110 web sites were scanned by these 6 agencies. The number of sites scanned per agency ranged from 11 to 27. Most sites were scanned weekly (41%) or monthly (33%). Thirty-one percent (31%) of the total number of sites was considered as highly important. The agencies spent at least 2 hours a week and at most 8 hours per week scanning the Internet. Although each agency's remit differed somewhat in scope, on average the same types of sites were scanned. These include sites from regulatory agencies, sites with information on new drugs or new devices, and sites with news from newswires. However, within these types there was not much correlation between the individual sites that agencies judged important to scan. CONCLUSION: The use of the Internet for identifying new health technologies is increasing in the majority of horizon scanning systems around the world. At the same time there is considerable variation between individual agencies in their approach to this source of information. This can only be partially explained by differences in scope of scanning activities of the individual agencies. A coordinated effort to develop Internet search strategies for either different categories of health technologies or different clinical specialties may improve efficiency and quality of scanning in terms of the number of potentially relevant technologies identified. PMID- 12746212 TI - Use of CD-ROM MEDLINE by medical students of the College of Medicine, University of Lagos, Nigeria. AB - BACKGROUND: Use of information technology in information acquisition, especially MEDLINE on CD-ROM and online, has been evaluated in several localities and regions, especially in the advanced countries. Use of MEDLINE on CD-ROM is still very poor among the medical students of the University of Lagos, Lagos, Nigeria, due to lack of awareness, insufficient personal computers, nonperiodic training, and the high cost of using the facility. Due to financial constraints, MEDLINE online and sufficiently-networked computer systems are not available. OBJECTIVE: To report on the situation in Nigeria, a developing country, so as to compare the current awareness of searching MEDLINE on CD-ROM among the medical students at the University of Lagos with the awareness of their overseas' counterparts. This is the first step toward setting up an online PubMed search as well as expanding the computer systems and network. METHODS: Essentially based on cross-sectional proportional sampling using structured questionnaires, in-depth interviews, and focus-group discussions among the medical students and library staff. The study involved the medical students in their second year to sixth (final) year of study. RESULTS: Of the 250 students interviewed, 130 (52%) were aware of MEDLINE on CD-ROM searches as a means of information retrieval. Only 60 (24%) had used MEDLINE on CD-ROM-2% had used MEDLINE on CD-ROM more than 9 times; 4%, 7 to 9 times; 8%, 4 to 6 times; and 10%, 1 to 3 times. Of the students who used MEDLINE on CD-ROM search, 22% used it in preparing for examinations, 24% in research, 6% in patient care, and 26% in preparation of assignments and clinical cases. Lack of awareness (52%) and cost of undertaking MEDLINE on CD-ROM search (46%) were identified as important factors that discouraged the use of MEDLINE on CD-ROM. CONCLUSION: Though the above factors were recognized as important, it was concluded that the reasons for the poor use of MEDLINE on CD-ROM are multifactorial. Poor use of MEDLINE on CD-ROM could be attributed to these critical underlying factors: nonavailability of networked personal computers, which should be connected to a central server; lack of mandatory assignments to the medical students that would specifically require use of MEDLINE on CD-ROM; financial constraints on the university management; and infrequent periodic orientation on use of MEDLINE on CD-ROM. It was therefore suggested that the number of personal computers should be increased and that the library staff should periodically train the preclinical and clinical medical students in searching MEDLINE on CD-ROM. These steps would enable the medical students to benefit from online PubMed searching when it becomes fully operational in the future. PMID- 12746213 TI - Alterations in lipid kinetics in men with HIV-dyslipidemia. AB - Hypertriglyceridemia is common in individuals with human immunodeficiency (HIV) infection, but the mechanisms responsible for increased plasma triglyceride (TG) concentrations are not clear. We evaluated fatty acid and VLDL-TG kinetics during basal conditions and during a glucose infusion that resulted in typical postprandial plasma glucose and insulin concentrations in six men with HIV dyslipidemia [body mass index (BMI): 28 +/- 2 kg/m2] and six healthy men (BMI: 26 +/- 2 kg/m2). VLDL-TG secretion and palmitate rate of appearance (Ra) in plasma were measured by using stable-isotope-labeled tracer techniques. Basal palmitate Ra and VLDL-TG secretion rates were greater (P < 0.01 for both) in men with HIV dyslipidemia (1.04 +/- 0.07 micromol palmitate x kg-1 x min-1 and 5.7 +/- 0.6 micromol VLDL-TG x l plasma-1 x min-1) than in healthy men (0.67 +/- 0.08 micromol palmitate. kg-1 x min-1 and 3.0 +/- 0.5 micromol VLDL-TG x l plasma-1 x min-1). Basal VLDL-TG plasma clearance was lower in men with HIV-dyslipidemia (13 +/- 1 ml/min) than in healthy men (19 +/- 2 ml/min; P < 0.05). Glucose infusion decreased palmitate Ra (by approximately 50%) and the VLDL-TG secretion rate (by approximately 30%) in both groups, but the VLDL-TG secretion rate remained higher (P < 0.05) in subjects with HIV-dyslipidemia. These findings demonstrate that increased secretion of VLDL-TG and decreased plasma VLDL-TG clearance, during both fasting and fed conditions, contribute to hypertriglyceridemia in men with HIV-dyslipidemia. Although it is likely that increased free fatty acid release from adipose tissue contributes to the increase in basal VLDL-TG concentration, other factors must be involved, because insulin-induced suppression of lipolysis and systemic fatty acid availability did not normalize the VLDL-TG secretion rate. PMID- 12746214 TI - Disruption of microtubules in rat skeletal muscle does not inhibit insulin- or contraction-stimulated glucose transport. AB - Insulin and muscle contractions stimulate glucose transport in skeletal muscle through a translocation of intracellular GLUT4 glucose transporters to the cell surface. Judged by immunofluorescence microscopy, part of the GLUT4 storage sites is associated with the extensive microtubule cytoskeleton found in all muscle fibers. Here, we test whether microtubules are required mediators of the effect of insulin and contractions. In three different incubated rat muscles with distinct fiber type composition, depolymerization of microtubules with colchicine for < or =8 h did not inhibit insulin- or contraction-stimulated 2-deoxyglucose transport or force production. On the contrary, colchicine at least partially prevented the approximately 30% decrease in insulin-stimulated transport that specifically developed during 8 h of incubation in soleus muscle but not in flexor digitorum brevis or epitrochlearis muscles. In contrast, nocodazole, another microtubule-disrupting drug, rapidly and dose dependently blocked insulin and contraction-stimulated glucose transport. A similar discrepancy between colchicine and nocodazole was also found in their ability to block glucose transport in muscle giant "ghost" vesicles. This suggests that the ability of insulin and contractions to stimulate glucose transport in muscle does not require an intact microtubule network and that nocodazole inhibits glucose transport independently of its microtubule-disrupting effect. PMID- 12746215 TI - Interactive effects of PTH and mechanical stress on nitric oxide and PGE2 production by primary mouse osteoblastic cells. AB - Parathyroid hormone (PTH) and mechanical stress both stimulate bone formation but have opposite effects on bone resorption. PTH increased loading-induced bone formation in a rat model, suggesting that there is an interaction of these stimuli, possibly at the cellular level. To investigate whether PTH can modulate mechanotransduction by bone cells, we examined the effect of 10-9 M human PTH-(1 34) on fluid flow-induced prostaglandin E2 (PGE2) and nitric oxide (NO) production by primary mouse osteoblastic cells in vitro. Mechanical stress applied by means of a pulsating fluid flow (PFF; 0.6 +/- 0.3 Pa at 5 Hz) stimulated both NO and PGE2 production twofold. In the absence of stress, PTH also caused a twofold increase in PGE2 production, but NO release was not affected and remained low. Simultaneous application of PFF and PTH nullified the stimulating effect of PFF on NO production, whereas PGE2 production was again stimulated only twofold. Treatment with PTH alone reduced NO synthase (NOS) enzyme activity to undetectable levels. We speculate that PTH prevents stress induced NO production via the inhibition of NOS, which will also inhibit the NO mediated upregulation of PGE2 by stress, leaving only the NO-independent PGE2 upregulation by PTH. These results suggest that mechanical loading and PTH interact at the level of mechanotransduction. PMID- 12746216 TI - Role and regulation of the fibroblast growth factor axis in human thyroid follicular cells. AB - Thyroidal levels of fibroblast growth factor-2 (FGF-2) and fibroblast growth factor receptor 1 (FGFR1) are elevated in human thyroid hyperplasia. To understand the significance of this, effects of FGFR1 activation on normal human thyrocyte growth and function in vitro and the regulation of FGF-2 and FGFR1 expression have been examined. FGF-2 stimulated cell growth, as measured by cell counting, and inhibited thyroid function as measured by 125I uptake. Sensitivity to FGF-2 disappeared after 7 days, although FGFR1 expression was maintained. Thyroid-stimulating hormone (TSH, 300 mU/l) increased FGFR1 mRNA expression within 4 h and protein expression by 8 h. Exogenous FGF-2 decreased FGFR1 protein. Endogenous FGF-2 levels were low (approximately 1-2 pg/microg protein), and TSH treatment decreased these by 50%. Protein kinase C (PKC) activation increased FGF-2 mRNA and FGF-2 secretion within 2 h. This effect was enhanced (4.4-fold) when cells were cultured in TSH. We conclude that TSH stimulates FGFR1 but not FGF-2 expression. PKC activation stimulates FGF-2 synthesis and secretion, and TSH synergizes with PKC activators. Increases in FGFR1 or FGF-2 or in both may contribute to goitrogenesis. PMID- 12746218 TI - Inhibition of lipopolysaccharide-inducible nitric oxide synthase, TNF-alpha and COX-2 expression by sauchinone effects on I-kappaBalpha phosphorylation, C/EBP and AP-1 activation. AB - 1. Sauchinone, a lignan isolated from Saururus chinensis (Saururaceae), is a diastereomeric lignan with cytoprotective and antioxidant activities in cultured hepatocytes. The effects of sauchinone on the inducible nitric oxide synthase (iNOS), tumor necrosis factor-alpha (TNF-alpha) and cyclooxygenase 2 (COX-2) gene expression and on the activation of transcription factors, nuclear factor-kappaB (NF-kappaB), CCAAT/enhancer-binding protein (C/EBP), activator protein-1 (AP-1) and cAMP-response element-binding protein (CREB) were determined in Raw264.7 cells as part of the studies on its anti-inflammatory effects. 2. Expression of the iNOS, TNF-alpha and COX-2 genes was assessed by Northern and Western blot analyses. NO production was monitored by chemiluminescence detection using a NO analyzer. To identify the transcriptional factors affected by sauchinone, the extents of NF-kappaB, C/EBP, AP-1 and CREB activation were measured. Activation of the transcription factors was monitored by gel mobility shift assay, whereas p65 and I-kappaBalpha were analyzed by immunocytochemical and immunoblot analyses. 3. Sauchinone inhibited the induction of iNOS, TNF-alpha and COX-2 by lipopolysaccharide (LPS) (IC500.05). 6. The tyrosine kinase inhibitor genistein (10-20 microM) blocked withdrawal hyper responsiveness. Of six EtOH-sensitive neurons, the mean NMDA-evoked current area after washout was 89+/-6% of control, P>0.05. 7 The protein kinase A (PKA) inhibitor Rp-cAMP (20-500 microM) did not block withdrawal hyper-responsiveness. On washout, the mean NMDA-evoked current area was 124+/-6% of control (n=5, P<0.05). 8 Two broad-spectrum specific protein kinase C (PKC) inhibitors, GF 109203X (0.3 microM) and chelerythrine chloride (0.5-2 nM), blocked withdrawal hyper-responsiveness. Responses on washout were 108+/-7%, n=5 and 88+/-4%, n=4 of control, respectively, P>0.05. 9 NMDA activation during EtOH exposure is necessary for withdrawal hyper-responsiveness. Both tyrosine kinase and PKC, but not PKA, appear to be essential for EtOH withdrawal hyper-responsiveness mediated by postsynaptic NMDA receptors in spinal cord motor neurons. PMID- 12746226 TI - Effects of the neutral endopeptidase inhibitor thiorphan on cardiovascular and renal function in cirrhotic rats. AB - 1. Cirrhosis is associated with cardiovascular and renal dysfunction including sodium retention. Many vasoactive peptides such as atrial natriuretic peptide (ANP) and endothelin-1 (ET-1) are degraded by neutral endopeptidase 24.11 (NEP). We investigated the hemodynamic and renal effects of thiorphan, a NEP inhibitor, in a rat cirrhosis model. 2. Cirrhosis was induced by chronic bile duct ligation, and controls had sham operation. Systemic and renal hemodynamics in conscious, restrained animals were determined using radiolabeled microspheres, and glomerular filtration rate (GFR) was measured by (3)H-inulin clearance. Plasma ANP and ET-1, and renal cGMP and Na(+) - K(+) ATPase activity were assayed. These variables were measured at baseline and after intravenous infusion of thiorphan (0.5 mg kg(-1) loading dose followed by 0.1 mg kg(-1) min(-1) x 30 min). 3. Thiorphan significantly decreased cardiac output, and increased systemic vascular resistance in controls, whereas in cirrhotic rats these variables were unchanged. 4. Compared to the controls, cirrhotic rats showed a decreased baseline GFR and urine sodium excretion, and the latter was significantly increased by thiorphan. 5. Thiorphan increased plasma ET-1 levels in controls, but not cirrhotic rats. ANP levels were not significantly increased in either group by thiorphan. 6. Thiorphan significantly increased cGMP concentrations and decreased Na(+) - K(+) ATPase activity of renal medulla but not cortex in cirrhotic rats; no effect was observed in the control rats. 7. We conclude that thiorphan induces natriuresis in cirrhotic rats by a direct renal medullary mechanism via cGMP and Na(+) - K(+) ATPase, without affecting systemic hemodynamics. This may potentially be useful in patients with ascites. PMID- 12746228 TI - SR33805, a Ca2+ antagonist with length-dependent Ca2+ -sensitizing properties in cardiac myocytes. AB - 1. This study examined the effects of SR33805, a fantofarone derivative with reported strong Ca(2+) -antagonistic properties, on the contractile properties of intact and skinned rat ventricular myocytes. 2. On intact cells loaded with the Ca(2+)-fluorescent indicator Indo-1, the application of low concentrations of SR33805 enhanced the amplitude of unloaded cell shortening and decreased the duration of cell shortening. Amplitude of the Ca(2+) transient was also decreased. 3. These effects were accompanied with a shortening of the action potential and a dose-dependent blockade of L-type calcium current (IC(50)=2.4 x 10(-8) M). 4. On skinned cardiac cells, the application of a low SR33805 concentration (10(-8) M) induced a significant increase in maximal Ca(2+) activated force at the two-tested sarcomere lengths (SLs), 1.9 and 2.3 microm. 5. The application of a larger dose of SR33805 (10(-6)-10(-5) M) induced a significant leftward shift of the tension-pCa relation that accounts for Ca(2+) sensitization of the myofilaments, particularly at 2.3 microm SL. 6. In conclusion, despite its strong Ca(2+)-antagonistic properties SR33805 increases cardiac cell contractile activity as a consequence of its Ca(2+)-sensitizing effects. These effects are attributable to both an increase in the maximal Ca(2+) activated force and a length-dependent Ca(2+)-sensitization. PMID- 12746227 TI - A novel rat lipoxin A4 receptor that is conserved in structure and function. AB - 1. Lipoxin (LX) A(4) and aspirin-triggered-LX (ATL) are endogenous lipid-derived mediators that regulate leukocyte trafficking via specific LXA(4) receptors (ALX), and are involved in endogenous anti-inflammation and resolution. Both LXA(4) and ATL are produced by rat tissues in vitro as well as in vivo. In rats, LXA(4) and ATL exhibit potent physiological and pathophysiological roles. Thus, we set out to determine whether ALX is expressed in rat tissues and its potential role in modulating leukocyte trafficking with LXA(4) and ATL. 2. In rats, a stable analog of ATL, when given intravenously with two consecutive doses at approximately 60 microg kg(-1) each injection, significantly inhibited neutrophil infiltration (approximately 43%) and protein extravasation (approximately 42%) in a casein-induced peritonitis. 3. The rat orthologue of ALX was cloned from peripheral blood leukocytes encoding a putative G protein-coupled receptor (GPCR). It gave approximately 74 and approximately 84% homology, respectively to the deduced amino-acid sequences of the human and mouse ALX. 4. Tissue distribution analysis by RNase protection revealed that this rat receptor is expressed in tissues/cells, where LXA(4) displays physiological and pathophysiological roles, namely, lung, kidney and leukocytes. 5. The rat orthologue of ALX gave specific radioligand binding with [(3)H]LXA(4) and [(125) Tyr]-annexin 1-derived peptide with apparent K(d) values of 5 and 820 nM, respectively, that are at levels comparable to those of the human ALX. 6. Activation of rat ALX inhibited tumor necrosis factor alpha-mediated nuclear factor kappaB activity in a ligand-dependent manner utilizing a luciferase reporter gene system. 7. Together, these results are the first demonstration of a rat ALX that is conserved in both structure and function suggesting that ALX plays key roles in regulating effector immune responses from murine to human species. PMID- 12746229 TI - Pharmacological characterisation of native somatostatin receptors in AtT-20 mouse tumour corticotrophs. AB - 1. The mouse corticotroph tumour cell line AtT-20 is a useful model to investigate the physiological role of native somatostatin (SRIF, Somatotropin release inhibitory factor) receptor subtypes (sst(1) - sst(5)). The objective of this study was to characterise the pharmacological features and the functional effects of SRIF receptors expressed by AtT-20 cells using radioligand binding and cAMP accumulation. 2. [(125)I]LTT-SRIF-28, [(125)I]CGP 23996, [(125)I]Tyr(10) cortistatin-14 and [(125)I]Tyr(3)-octreotide labelled SRIF receptor binding sites with high affinity and in a saturable manner (B(max)=315, 274, 239 and 206 fmol mg(-1), respectively). [(125)I]LTT-SRIF-28 labels significantly more sites than [(125)I]Tyr(10) -cortistatin-14 and [(125)I]Tyr(3) -octreotide as seen previously in cells expressing pure populations of sst(2) or sst(5) receptors. 3. SRIF analogues displaced the binding of the four radioligands. sst(2/5) receptor selective ligands showed much higher affinity than sst(1/3/4) receptor-selective ligands. The binding profile of [(125)I]Tyr(3)-octreotide was different from that of [(125)I]LTT-SRIF-28, [(125)I]CGP 23996 and [(125)I]Tyr(10)-cortistatin-14. The sst(5/1) receptor-selective ligand L-817,818 identified two binding sites, one with subnanomolar affinity (sst(5) receptors) and one with micromolar affinity (sst(2) receptors); however, the proportions were different: 70 - 80% high affinity with [(125)I]LTT-SRIF-28, [(125)I]CGP 23996, [(125)I]Tyr(10)-cortistatin 14, but only 20% with [(125)I]Tyr(3)-octreotide. 4. SRIF analogues inhibited the forskolin-stimulated cAMP levels depending on concentration. sst(2/5) receptor selective ligands were highly potent, whereas sst(1/3/4) receptor-selective ligands had no significant effects. The sst(2) receptor antagonist D-Tyr(8)-CYN 154806 competitively antagonised the effects of SRIF-14 and sst(2) receptor preferring agonists, but not those of L-817,818. 5. The complex binding properties of SRIF receptor analogues indicate that sst(2) and sst(5) receptors are the predominant SRIF receptors expressed on AtT-20 cell membranes with no or only negligible presence of sst(1), sst(3) and sst(4) receptors. In the functional studies using cAMP accumulation, only sst(2) and sst(5) receptors appear to play a role. However, the "predominant" receptor appears to be the sst(2) receptor, although sst(5) receptors can also mediate the effect, when the ligand is not able to activate sst(2) receptors. This clearly adds flexibility to SRIF-mediated functional effects and suggests that the physiological role of SRIF and its analogues may be mediated preferentially via one subtype over another. PMID- 12746230 TI - Different molecular sites of action for the KATP channel inhibitors, PNU-99963 and PNU-37883A. AB - 1. We investigated the mechanism of action of two novel nonsulphonylurea ATP sensitive potassium channel (K(ATP)) inhibitors, PNU-99963 and PNU-37883A, on four types of cloned K(ATP) channels. 2. Whole-cell currents were recorded in a symmetrical potassium (140 mM) gradient in HEK-293 cells stably expressing Kir6.2/SUR1, Kir6.2/SUR2A, Kir6.2/SUR2B or Kir6.1/SUR2B. 3. PNU-99963 potently inhibited the four K(ATP) channel clones. The concentration at which half-maximum current was inhibited (IC(50)) was 66, 41, 43 and 11 nM for Kir6.2/SUR1, Kir6.2/SUR2A, Kir6.2/SUR2B and Kir6.1/SUR2B, respectively. In contrast, PNU-99963 up to a concentration of 3 microM had no significant effect on current generated in HEK-293 cells by transiently expressing Kir6.2Delta26, a C-terminal truncated pore-forming subunit of Kir6.2. 4. PNU-37883A inhibited four types of K(ATP) channels, but to different extents. Inhibition of the putative smooth muscle K(ATP) channel types, Kir6.2/SUR2B (IC(50); 15 microM) and Kir6.1/SUR2B (IC(50); 6 microM), was significantly greater than inhibition of either the pancreatic beta cell or cardiac K(ATP) channel clones. Moreover, PNU-37883A significantly inhibited currents generated by expressing Kir6.2Delta26 alone, with an IC(50) of 5 microM, which was significantly increased to 38 microM when Kir6.2Delta26 was expressed with SUR2B. 5. In conclusion, two structurally different nonsulphonylurea compounds, PNU-99963 and PNU-37883A, inhibit K(ATP) channels via different mechanisms, namely through the sulphonylurea receptor (SUR) and the pore-forming subunits, respectively, although SUR2B reduced the inhibitory effect of PNU-37883A. While PNU-99963 potently inhibits all the four cloned K(ATP) channels, PNU-37883A has a degree of selectivity towards both smooth muscle K(ATP) channels, but could not discriminate between them. PMID- 12746231 TI - Role of the bradykinin B2 receptor for the local and systemic inflammatory response that follows severe reperfusion injury. AB - 1. Bradykinin (BK) appears to play an important role in the development and maintenance of inflammation. Here, we assessed the role of the BK B(2) receptor for the injuries that occur after ischemia and reperfusion (I/R) of the territory irrigated by the superior mesenteric artery. 2. Tissue (lung and duodenum) kallikrein activity increased after ischemia with greater enhancement after reperfusion. A selective inhibitor of tissue kallikrein, Phenylacetyl-Phe-Ser-Arg N-(2,3-dinitrophenyl)-ethylenediamine (TKI, 0.001-10 mg ml(-1)), inhibited kallikrein activity in a concentration-dependent manner in vitro. In vivo, pretreatment with TKI (30 mg kg(-1)) prevented the extravasation of plasma and the recruitment of neutrophils. 3. Similarly, the bradykinin B(2) receptor antagonists, HOE 140 (0.01-1.0 mg kg(-1)) or FR173657 (10.0 mg kg(-1)), inhibited reperfusion-induced increases in vascular permeability and the recruitment of neutrophils in the intestine and lungs. 4. In a model of more severe I/R injury, HOE 140 (1.0 mg kg(-1)) inhibited the increase in vascular permeability, neutrophil recruitment, haemorrhage and tissue pathology. Furthermore, HOE 140 significantly inhibited the elevations of TNF-alpha in tissue and serum and partially prevented lethality. This was associated with an increase in the concentrations of IL-10 in tissue and serum. 5. Thus, our results demonstrate that, following intestinal I/R injury, there is an increase in tissue kallikrein activity and activation of BK B(2) receptors. B(2) receptor activation is essential for the development of inflammatory tissue injury and lethality. These results contrast with those of others showing that BK mostly exerts a protective role during I/R injury. PMID- 12746232 TI - YM-53601, a novel squalene synthase inhibitor, suppresses lipogenic biosynthesis and lipid secretion in rodents. AB - 1. To better understand how it decreases plasma cholesterol and triglyceride levels, we evaluated the effect of (E)-2-[2-fluoro-2-(quinuclidin-3 ylidene)ethoxy]-9H-carbazole monohydrochloride(YM-53601) on lipogenic biosynthesis in the liver and lipid secretion from the liver in rats and hamsters. 2. Single administration of YM-53601 in cholestyramine-treated rats inhibited triglyceride and free fatty acid (FFA) biosynthesis at a similar dose range to that at which it inhibited cholesterol biosynthesis. YM-53601 inhibited both triglyceride and FFA biosynthesis in hamsters treated with cholestyramine. 3. YM-53601 by single oral administration decreased the enhanced plasma triglyceride levels in hamsters induced by an injection of protamine sulfate, which inhibits lipoprotein lipase (LPL) and consequently increases plasma very low-density lipoprotein (VLDL) triglyceride levels. YM-53601 also decreased the enhanced plasma triglyceride and cholesterol levels in hamsters treated with Triton WR1339, which also inhibits the degradation of VLDL. Plasma cholesterol was significantly decreased as soon as 1 h after single administration of YM 53601 in hamsters fed a normal diet. 4. This is the first report that a squalene synthase inhibitor suppresses lipogenic biosynthesis in the liver and cholesterol and triglyceride secretion from the liver in vivo. We therefore suggest that the mechanism by which YM-53601 decreases plasma triglyceride might include these effects. The finding that YM-53601 rapidly decreased plasma cholesterol suggests that this compound may be effective in decreasing plasma cholesterol levels early in the course of treatment of hypercholesterolemia in humans. PMID- 12746233 TI - Actions of LY341495 on metabotropic glutamate receptor-mediated responses in the neonatal rat spinal cord. AB - 1. The group II metabotropic glutamate (mGlu) receptor antagonist (2S,1'S,2'S)-2 (2-carboxycyclopropyl)-2-(9H-xanthen-9-yl)glycine (LY341495) also has activity at group I and III mGlu receptors at higher concentrations and can be used to discriminate between mGlu receptor subtypes. We report the antagonist action of LY341495 on glutamate receptors expressed in the neonatal rat spinal cord preparation and the use of this antagonist to investigate the group III mGlu receptor subtypes responsible for mediating the depression of synaptic transmission in the spinal cord mediated by the group III mGlu receptor agonists (S)-2-amino-4-phosphonobutanoic acid ((S)-AP4) and (1S,3R,4S)-1-aminocyclopentane 1,2,4-tricarboxylic acid (ACPT-I). 2. LY341495 antagonised mGlu receptor agonist induced responses in the spinal cord with a rank order of potency of group II > group III > group I, which is the same as that observed in human cloned mGlu receptor cell lines. Antagonism of group II and III mGlu receptor-mediated effects were time dependent when low-nanomolar concentrations of LY341495 were used. Although the rank order of potency of LY341495 was the same on native rat and cloned human mGlu receptors, there was a compression in the selectivity between group II and III mGlu receptors, expressed in the spinal cord. 3. In agreement with a previous study on cloned ionotropic glutamate receptors 100 microM LY341495 had little or no effect on N-methyl-D-aspartate, (S)-2-amino-3-(3 hydroxy-5-methylisoxazol-4-yl) propionic acid or kainate receptor-mediated responses on motoneurones. 4. LY341495 exhibited low-nanomolar potency antagonist activity against (S)-AP4 and ACPT-I suggesting that these agonists are activating predominantly mGlu8 and that mGlu4 receptors do not play a role in modulating synaptic transmission in the pathways stimulated in the experiments described here. PMID- 12746234 TI - Enhancement of alpha2-adrenoceptor-mediated vasoconstriction by the thromboxane mimetic U46619 in the porcine isolated ear artery: role of the ERK-MAP kinase signal transduction cascade. AB - 1. Alpha(2)-adrenoceptor-mediated contractions in porcine blood vessels can be enhanced in the presence of the thromboxane-mimetic U46619, and forskolin. The aim of this study was to determine the role of U46619 in the enhanced contractions, and to determine whether signalling through the ERK-MAP kinase pathway is involved. 2. Responses to the alpha(2)-adrenoceptor agonist UK14304 (1 micro M) were increased from 22+/-3% of the response to 60 mM KCl to 68+/-12% (n=8, mean+/-s.e.m.) in the presence of a low concentration of U46619 (< 20% of the 60 mM KCl response). 3. Both the direct and the U46619-enhanced UK14304 responses were inhibited by 50 microM PD98059, an inhibitor of the ERK-MAP kinase pathway. UK14304-induced contractions were associated with an increase in ERK2 phosphorylation, indicating an increased activity. In the presence of U46619, there was an enhanced phosphorylation of ERK2. U46619 on its own had no effect on ERK phosphorylation. 4. Both the direct and enhanced UK14304 contractions were inhibited in the absence of extracellular calcium. These conditions also prevented the increase in ERK2 phosphorylation. This indicates a role for calcium influx in the enhanced contractions. 5. In conclusion, this study demonstrates that precontraction with the thromboxane-mimetic U46619 enhances alpha(2) adrenoceptor-mediated vasoconstriction through the enhancement of the ERK-MAP kinase pathway, and influx of extracellular calcium. PMID- 12746235 TI - PPAR-alpha and -gamma but not -delta agonists inhibit airway inflammation in a murine model of asthma: in vitro evidence for an NF-kappaB-independent effect. AB - 1. Peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors that have been proposed to regulate inflammation by antagonising the nuclear factor-kappaB (NF-kappaB) signalling pathway. We investigated the role of PPARs using synthetic agonists in murine models of airway inflammation, and addressed the possible effect on NF-kappaB signalling in vitro using a human epithelial cell line, A549. 2. Sensitised BALB/c mice exposed to an aerosol solution of ovalbumin had an increased number of airway eosinophils, neutrophils and lymphocytes. When given intranasally an hour before the aerosol challenge, a PPAR-alpha (GW 9578) and PPAR-gamma (GI 262570) selective agonist as well as a dual PPAR-alpha/gamma (GW 2331) agonist selectively inhibited allergen-induced bronchoalveolar lavage eosinophil and lymphocyte but not neutrophil influx. In contrast, a PPAR-delta agonist (GW 501516) was inactive. 3. When given intranasally an hour before challenge, PPAR alpha and PPAR-gamma selective agonists as well as a dual PPAR-alpha/gamma agonist did not inhibit lipopolysaccharide-induced bronchoalveolar lavage neutrophil influx or tumour necrosis factor-alpha (TNF-alpha) and KC production. 4. In A549 cells, selective agonists for PPAR-alpha, -gamma and -delta did not inhibit intracellular adhesion molecule-1 expression following stimulation with proinflammatory cytokines. In addition, IL-8 release and the activation of an NF kappaB-responsive reporter gene construct were inhibited only at micromolar concentrations, suggesting that these effects were not PPAR-mediated. 5. Our in vivo data show that agonists of PPAR-alpha and -gamma, but not -delta, inhibit allergen-induced bronchoalveolar lavage eosinophil and lymphocyte influx. In vitro data suggest that this effect might not be mediated by antagonism of the NF kappaB pathway. PMID- 12746236 TI - Alteration of the purinergic modulation of enteric neurotransmission in the mouse ileum during chronic intestinal inflammation. AB - 1. The effect of chronic intestinal inflammation on the purinergic modulation of cholinergic neurotransmission was studied in the mouse ileum. Chronic intestinal inflammation was induced by infection of mice with the parasite Schistosoma mansoni during 16 weeks. 2. S. mansoni infection induced a chronic inflammatory response in the small intestine, which was characterised by intestinal granuloma formation, increased intestinal wall thickness, blunted mucosal villi and an enhanced activity of myeloperoxidase. 3. In control ileum and in chronically inflamed ileum, electrical field stimulation (EFS) of longitudinal muscle strips induced frequency-dependent contractions that were abolished by tetrodotoxin (TTX) and atropine. Carbachol induced dose-dependent contractions that were not affected by TTX but abolished by atropine. 4. In control ileum, adenosine and ATP dose-dependently inhibited the contractions to EFS. Theophylline and 8 phenyltheophylline, P(1) and A(1) receptor antagonists respectively, prevented this inhibitory effect of adenosine and ATP. PPADS, DMPX and MRS 1220, antagonists of P(2), A(2) and A(3) receptors, respectively, did not prevent this inhibitory effect of adenosine and ATP. Adenosine and ATP did not affect the contractions to carbachol. 5. The inhibitory effect of adenosine and ATP on contractions to EFS in control ileum was mimicked by the stable adenosine analogue methyladenosine and by the A(1)-receptor agonist N(6) cyclohexyladenosine, but not by the A3 receptor agonist 2-Cl IB-MECA or by the ATP analogues alphabeta-methylene-ATP and ADPbetaS. The inhibitory effect of adenosine on contractions to EFS was lost after prolonged (90 min) treatment of control ileum with methyladenosine (100 micro M). 6. In chronically inflamed ileum, adenosine, methyladenosine, N(6)-cyclohexyladenosine and ATP all failed to inhibit the cholinergic nerve-mediated contractions to EFS. Also theophylline, 8 phenyltheophylline, PPADS, DMPX and MRS 1220 had no effect on the contractions to EFS and carbachol. The loss of effect of adenosine and ATP was still evident after 52 weeks of infection. 7. These results indicate that in physiological conditions neuronal adenosine A(1) receptors modulate cholinergic nerve activity in the mouse ileum. However, during chronic intestinal inflammation, this purinergic modulation of cholinergic nerve activity is impaired. This suggests that chronic intestinal inflammation leads to a dysfunction of specific neuronal regulatory mechanisms in the enteric nervous system. PMID- 12746237 TI - Environmental tobacco smoke and absenteeism related to respiratory illness in schoolchildren. AB - Household environmental tobacco smoke (ETS) exposure accounts for substantial morbidity among young children, but the ETS-associated morbidity burden among school-age children is less well defined. Illness-related school absenteeism is a measure of a broad spectrum of adverse effects of ETS exposure in school-age children. The authors investigated the relations between ETS exposure, asthma status, and illness-related school absenteeism in a cohort of 1,932 fourth-grade schoolchildren from 12 southern California communities during January-June 1996. Incidence rates and adjusted relative risks of illness-related absences were determined by using an active surveillance system. The effects of ETS exposure on absenteeism were assessed by using stratified incidence rates and Poisson regression to adjust for sociodemographic factors. ETS exposure was associated with an increased risk of respiratory-illness-related school absences (relative risk (RR) = 1.27, 95% confidence interval (CI): 1.04, 1.56). Children living in a household with two or more smokers were at increased risk of such absences (RR = 1.75, 95% CI: 1.33, 2.30). Children's asthma status affected their response to ETS. Compared with unexposed children without asthma, children with asthma were at increased risk of respiratory-illness-related school absences when exposed to one (RR = 2.35, 95% CI: 1.49, 3.71) or two or more (RR = 4.45, 95% CI: 2.80, 7.07) household smokers. Children without asthma also had an increased risk if exposed to two or more smokers (RR = 1.44, 95% CI: 1.04, 2.00). Therefore, ETS exposure is associated with increased respiratory-related school absenteeism among children, especially those with asthma. PMID- 12746238 TI - Invited commentary: Attendance and absence as markers of health status--the example of active and passive cigarette smoking. PMID- 12746239 TI - Age, flight experience, and risk of crash involvement in a cohort of professional pilots. AB - Federal aviation regulations prohibit airline pilots from flying beyond the age of 60 years. However, the relation between pilot age and flight safety has not been rigorously assessed using empirical data. From 1987 to 1997, the authors followed a cohort of 3,306 commuter air carrier and air taxi pilots who were aged 45-54 years in 1987. During the follow-up period, the pilots accumulated a total of 12.9 million flight hours and 66 aviation crashes, yielding a rate of 5.1 crashes per million pilot flight hours. Crash risk remained fairly stable as the pilots aged from their late forties to their late fifties. Flight experience, as measured by total flight time at baseline, showed a significant protective effect against the risk of crash involvement. With adjustment for age, pilots who had 5,000-9,999 hours of total flight time at baseline had a 57% lower risk of a crash than their less experienced counterparts (relative risk = 0.43, 95% confidence interval: 0.21, 0.87). The protective effect of flight experience leveled off after total flight time reached 10,000 hours. The lack of an association between pilot age and crash risk may reflect a strong "healthy worker effect" stemming from the rigorous medical standards and periodic physical examinations required for professional pilots. PMID- 12746240 TI - Alcohol concentration and risk of oral cancer in Puerto Rico. AB - Alcohol consumption is a major risk factor for cancers of the mouth and pharynx (oral cancer), but the differential risks by beverage type are unclear. In this 1992-1995 study, the authors examined oral cancer risk in Puerto Rico, comparing alcohol intake among 286 male cases aged 21-79 years and 417 population-based male controls, frequency matched by age. Heavy consumers of liquor (>/=43 drinks per week) had strongly increased risks of oral cancer (odds ratio = 6.4, 95% confidence interval: 2.4, 16.8); beer/wine showed only modest effects. Among liquor drinkers, risks were consistently greater for those who drank straight (undiluted) liquor than for those who usually drank mixed (diluted) liquor (odds ratio = 4.0, 95% confidence interval: 2.4, 6.7). Risks associated with combined exposure to tobacco were also more pronounced when subjects drank liquor straight. The elevated risks associated with drinking homemade rum were similar to those for other types of liquor. These results suggest that alcohol concentration is a risk factor for oral cancer independent of the total quantity of alcohol consumed. PMID- 12746241 TI - Factors associated with dietary supplement use among healthy adults of five ethnicities: the Multiethnic Cohort Study. AB - Participants of the Multiethnic Cohort Study in Hawaii and Los Angeles, California, a representative sample of African-American, Native Hawaiian, Latino, Japanese-American, and White adults, completed a baseline questionnaire in 1993 1996 assessing dietary supplement use during the past year as well as demographic, dietary, and other lifestyle factors. Factors associated with supplement use were examined among those who reported an absence of chronic disease (n = 100,196). Use of any of eight supplements at least once per week during the past year ranged from 44% among Hawaiian men to 75% among Japanese American and White women. Multivitamins were the most frequently reported supplement; 48% of the men and 56% of the women reported regular use. Dietary supplement use was high across all ethnic groups, although levels and length of regular use varied. In all gender-specific ethnic groups, supplement use tended to increase with age, education, physical activity, fruit intake, and dietary fiber intake and to decrease with obesity, smoking, and dietary fat intake. Participants whose lifestyles were healthier were more likely to use dietary supplements. Therefore, it may be difficult to separate the effects of supplement use from other lifestyle factors when studying disease etiology. PMID- 12746242 TI - Spatial clustering of amyotrophic lateral sclerosis in Finland at place of birth and place of death. AB - Previous evidence for spatial clustering of amyotrophic lateral sclerosis is inconclusive. Studies that have identified apparent clusters have often been based on a small number of cases, which means the results may have occurred by chance processes. Also, most studies have used the geographic location at the time of death as the basis for cluster detection, rather than exploring clusters at other points in the life cycle. In this study, the authors examine 1,000 cases of amyotrophic lateral sclerosis distributed throughout Finland who died between June 1985 and December 1995. Using a spatial-scan statistic, the authors examine whether there are significant clusters of the disease at both time of birth and time of death. Two significant, neighboring clusters were identified in southeast and south-central Finland at the time of death. A single significant cluster was identified in southeast Finland at the time of birth, closely matching one of the clusters identified at the time of death. These results are based on a large sample of cases, and they provide convincing evidence of spatial clustering of this condition. The results demonstrate also that, if the cluster analysis is conducted at different stages of the cases' life cycle, different conclusions about where potential risk factors may exist might result. PMID- 12746243 TI - Risk factors for lower urinary tract symptoms in a population-based sample of African-American men. AB - Previous epidemiologic studies evaluating risk factors for lower urinary tract symptoms (LUTS) have focused on White populations. Between September 1996 and January 1998, in a population-based sample of African-American men aged 40-79 years in Flint, Michigan, the authors assessed the role of putative sociodemographic, lifestyle, and medical history risk factors in moderate to severe LUTS, including the subcategories of obstructive and irritative symptoms. After the exclusion of men with prostate cancer or prior prostate surgery and men who were taking alpha-blockers for urinary tract symptoms, 708 participants provided responses to a structured interviewer-administered questionnaire. After multivariable adjustment, current and former smokers were at increased risk of moderate to severe LUTS, including obstructive symptoms. Heavy alcohol consumption and a history of hypertension or diabetes were positively associated with LUTS, and high income (>/=$30,000) was inversely associated with LUTS and with obstructive and irritative symptoms. A history of heart disease was positively associated with LUTS and with irritative symptoms. To the authors' knowledge, this was the first population-based study undertaken in African American men to evaluate putative risk factors for moderate to severe LUTS, including subcategories of obstructive and irritative urinary symptoms. These results describe associations with specific lifestyle and medical history risk factors. PMID- 12746244 TI - Trends in human immunodeficiency virus seroincidence among street-recruited injection drug users in San Francisco, 1987-1998. AB - This paper reports trends in human immunodeficiency virus (HIV) incidence among street-recruited injection drug users (IDUs) in San Francisco, California, from 1987 through 1998, estimated using a sensitive/less sensitive enzyme immunoassay testing strategy. IDUs were enrolled in 23 semiannual cross-sections from three community sites. For identification of recent infections, less sensitive enzyme immunoassay testing was performed on stored specimens that had previously tested positive for HIV antibodies. Annualized incidence rates were calculated and logistic regression models were fitted for assessment of risk factors for recent HIV infection. Among 8,065 susceptible IDUs, 34 had recent infections, for an incidence rate of 1.2% (95% confidence interval: 0.7, 2.0) per person-year. This rate declined from 2.7% in 1987/1988 to approximately 1% per person-year between 1989 and 1998. Two IDU subpopulations were at highest risk for infection: persons under age 30 years (2.8% per person-year) and men who had sex with men (3.0% per person-year). Participants who reported prior HIV test-result counseling were less likely than others to become infected (adjusted odds ratio = 0.43, 95% confidence interval: 0.21, 0.87). Sensitive/less sensitive enzyme immunoassay testing is an effective tool for assessing HIV incidence. HIV incidence among street-recruited IDUs in San Francisco appears to have remained stable and moderate since the late 1980s. PMID- 12746245 TI - Age at natural menopause and all-cause mortality: a 37-year follow-up of 19,731 Norwegian women. AB - In a cohort of 19,731 Norwegian postmenopausal women, the authors analyzed relations between the age at natural menopause and all-cause mortality. A total of 18,533 women died during the 37 years of follow-up from 1961 to 1997. An inverse relation was found between the age at menopause and the all-cause mortality rate (p = 0.003). The strength of the association was moderate, however, with 1.6% (95% confidence interval: 0.6, 2.7) reduced mortality per 3 years' increase in age at menopause. The impact appeared to be stronger in women with an attained age of less than 70 years (3.7% reduction in risk) than in women aged 80 years or more (1.0%). The inverse relation could not be explained by extreme mortality rates in women with very early (<40 years) or late (>55 years) menopause or by possible confounding variables like birth cohort, place of residence, occupational category (own or husband's occupation), body mass index, age at menarche, and first and last delivery or parity. The smoking prevalence was low in the underlying population, and the use of hormone replacement therapy was very rare. The authors conclude that age at natural menopause is inversely related to all-cause mortality. PMID- 12746246 TI - Sociodemographic and behavioral characteristics associated with timeliness and retention in a 6-month follow-up study of high-risk injection drug users. AB - Timeliness and retention in a 6-month follow-up study were analyzed by subjects' baseline characteristics in a seroincidence study of 263 injection drug users at high risk of human immunodeficiency virus infection. Subjects were recruited from September 1997 to June 1998 in community settings in Philadelphia, Pennsylvania. Of these subjects, 93% were completers: 11% before the targeted date, 38% at the targeted date, 32% within 1 month of delay, and 12% beyond 1 month. Late completers were more likely than other completers to be younger and to live farther away from the study center, less likely to have stayed in a shelter or a welfare residence during the past year, more likely to have a lower income, and more likely to have shared rinse water, cotton, or cooker. By contrast, loss to follow-up was not associated with these variables. Subjects lost to follow-up were more likely than those retained to have a high school diploma and to have moved during the past year; their source of needles was less likely to be a needle exchange program and more likely to be a shooting gallery. None of the drug-related behaviors that increase the risk of human immunodeficiency virus infection was associated with timeliness or retention, suggesting that the study might be minimally biased. PMID- 12746247 TI - Estimating the relative risk in cohort studies and clinical trials of common outcomes. AB - Logistic regression yields an adjusted odds ratio that approximates the adjusted relative risk when disease incidence is rare (<10%), while adjusting for potential confounders. For more common outcomes, the odds ratio always overstates the relative risk, sometimes dramatically. The purpose of this paper is to discuss the incorrect application of a proposed method to estimate an adjusted relative risk from an adjusted odds ratio, which has quickly gained popularity in medical and public health research, and to describe alternative statistical methods for estimating an adjusted relative risk when the outcome is common. Hypothetical data are used to illustrate statistical methods with readily accessible computer software. PMID- 12746248 TI - Reliability and validity of self-report of vitamin and mineral supplement use in the vitamins and lifestyle study. AB - In the United States, dietary supplements contribute a large proportion of micronutrient intakes. Therefore, it is important to collect accurate information on supplement use for studies of micronutrients and disease risk. This report describes the test-retest reliability and validity of a detailed, self administered mailed questionnaire on vitamin and mineral supplement use. Participants (n = 220) completed the questionnaire at baseline and 3 months later. During an in-person interview, participants provided spot urine and blood samples, and interviewers transcribed nutrient information from their supplement bottle labels. The questionnaire had very good test-retest reliability for mean supplement intake over the past 10 years, with intraclass correlations ranging from 0.69 for beta-carotene to 0.87 for vitamin E. Pearson's correlation coefficients comparing current supplemental intakes from the questionnaire and interviews/label transcriptions were high, ranging from 0.58 for beta-carotene to 0.82 for chromium; however, for some nutrients, median intakes from the questionnaire were slightly lower than from the interviews. Beta-carotene, vitamin C, and vitamin E (alpha-tocopherol) showed clear linear trends of increasing blood concentrations with higher self-reported supplemental intakes (Pearson's correlation coefficients adjusted for potential confounding factors and diet = 0.31, 0.29, and 0.69, respectively; all p < 0.0001). Creatinine adjusted spot urinary calcium values were not associated with supplemental calcium intakes (Pearson's r = -0.07). This self-administered questionnaire demonstrated high reproducibility and validity for collecting detailed information on supplement use. PMID- 12746250 TI - Re: "Estimation of risk of cancers before occurrence of acquired immunodeficiency syndrome in persons infected with human immunodeficiency virus". PMID- 12746251 TI - Identification of upper airway anatomic risk factors for obstructive sleep apnea with volumetric magnetic resonance imaging. AB - We used sophisticated volumetric analysis techniques with magnetic resonance imaging in a case-control design to study the upper airway soft tissue structures in 48 control subjects (apnea-hypopnea index, 2.0 +/- 1.6 events/hour) and 48 patients with sleep apnea (apnea-hypopnea index, 43.8 +/- 25.4 events/hour). Our design used exact matching on sex and ethnicity, frequency matching on age, and statistical control for craniofacial size and visceral neck fat. The data support our a priori hypotheses that the volume of the soft tissue structures surrounding the upper airway is enlarged in patients with sleep apnea and that this enlargement is a significant risk factor for sleep apnea. After covariate adjustments the volume of the lateral pharyngeal walls (p < 0.0001), tongue (p < 0.0001), and total soft tissue (p < 0.0001) was significantly larger in subjects with sleep apnea than in normal subjects. These data also demonstrated, after covariate adjustments, significantly increased risk of sleep apnea the larger the volume of the tongue, lateral pharyngeal walls, and total soft tissue: (1) total lateral pharyngeal wall (odds ratio [OR], 6.01; 95% confidence interval [CI], 2.62-17.14); (2) total tongue (OR, 4.66; 95% CI, 2.31-10.95); and (3) total soft tissue (OR, 6.95; 95% CI, 3.08-19.11). In a multivariable logistic regression analysis the volume of the tongue and lateral walls was shown to independently increase the risk of sleep apnea. PMID- 12746252 TI - Composite spirometric-computed tomography outcome measure in early cystic fibrosis lung disease. AB - With the advent of therapies aimed at young patients with cystic fibrosis, who have mildly reduced pulmonary function, the need for improved outcome measures that discriminate treatment effects has become important. Pulmonary function measurements or chest high-resolution computed tomography (HRCT) scores have been separately used to assess interventions. We evaluated these modalities separately and together during a treatment study to develop a more sensitive outcome measure. In a 1-year trial, 25 children randomized either to daily Pulmozyme or to normal saline aerosol were evaluated at randomization and at 3 and 12 months. Outcome variables were pulmonary function test (PFT) results, a global HRCT score, and a composite score incorporating PFTs and HRCT scoring. Regression analyses with generalized estimating equations permitted estimation of the difference in treatment effect between groups over time for each outcome. The largest difference in treatment effects observed at 12 months, measured by the percentage change from baseline, were with the composite total and maximal CT/PFT scores (35.4 and 30.4%), compared with mean forced expiratory flow during the middle half of the FVC (FEF25-75%) (13.0%) and total and maximal global HRCT scores (6.2%, 7.2%). The composite total and maximal CT/PFT scores were the most sensitive outcome measures for discriminating a treatment effect in children with cystic fibrosis with normal or mildly reduced pulmonary function during a 1-year trial of Pulmozyme. PMID- 12746253 TI - Pneumococcal septic shock is associated with the interleukin-10-1082 gene promoter polymorphism. AB - Polymorphisms in the tumor necrosis factor and interleukin-10 genes, linked to cytokine inducibility, may influence the inflammatory response to infection. We studied the biallelic interleukin-10-1082 promoter, the tumor necrosis factor alpha-308 promoter, and the lymphotoxin-alpha polymorphisms with regard to the development of septic shock in pneumococcal infection. Sixty-nine patients with pneumococcal disease (61 patients with community-acquired pneumonia, 5 patients with meningitis, and 3 patients with pneumonia and meningitis) and 50 age-matched control subjects were included. The polymorphisms were determined by the polymerase chain reaction. In patients with pneumococcal disease, the lipopolysaccharide-stimulated tumor necrosis factor and interleukin-10 release from whole blood were measured by ELISA. Sepsis severity was documented according to standard criteria. No significant genotypic differences were seen between patients and control subjects. Thirteen of 69 patients with pneumococcal disease developed septic shock. Interleukin-10 allele G homozygous patients had the highest risk for septic shock (odds ratio of 6.1; 95% confidence interval, 1.4 27.2; corrected p = 0.024). The stimulated interleukin-10 release was highest in interleukin-10 G homozygous patients (p = 0.04). In conclusion, interleukin-10 polymorphism, associated with high interleukin-10 inducibility, might influence the outcome of pneumococcal infection via induced immunosuppression and impaired bacterial clearance. PMID- 12746254 TI - Transforming growth factor-beta1 gene polymorphisms are associated with disease progression in idiopathic pulmonary fibrosis. AB - Transforming growth factor-beta1 (TGF-beta1) is a cytokine that plays a key role in the development of idiopathic pulmonary fibrosis. There have been reports on the presence of two genetic polymorphisms in the DNA sequence encoding the leader sequence of the TGF-beta1 protein, located in codons 10 and 25. The objective of this study was to investigate the association between TGF-beta1 gene polymorphisms in codons 10 and 25 and the susceptibility to idiopathic pulmonary fibrosis and the progression of the disease. Compared with healthy control subjects (n = 140), patients with idiopathic pulmonary fibrosis (n = 128) showed no significant deviations in genotype or allele frequencies. One hundred and ten patients with idiopathic pulmonary fibrosis were followed up for 30.3 +/- 25 months. The presence of a proline allele at codon 10 was independently associated with a significant increase in alveolar arterial oxygen tension difference during follow-up, after controlling for the effect of treatment (coefficient = 0.59; 95% confidence intervals, 0.23 to 0.96; p = 0.002). These findings suggest that (1) TGF-beta1 gene polymorphisms in codons 10 and 25 do not predispose to the development of idiopathic pulmonary fibrosis; and (2) TGF-beta1 gene polymorphisms may affect disease progression in patients with idiopathic pulmonary fibrosis. PMID- 12746255 TI - Use of isoniazid for latent tuberculosis infection in a public health clinic. AB - Isoniazid is an efficacious treatment for latent tuberculosis. Concerns remain, however, regarding hepatotoxicity associated with this medication. In addition, adherence may be suboptimal because at least 6 months of treatment is required. We extracted information from our latent tuberculosis treatment database to determine adverse effects and treatment completion rates associated with the use of isoniazid at a county tuberculosis clinic. Outcomes were available for 3,788 patients started on isoniazid between 1999 and 2002. Six hundred seventy-two patients (18%) experienced one or more adverse effects, including 10 (0.3%) determined to have isoniazid-associated liver injury. No hospitalizations or deaths occurred in patients experiencing an adverse effect. A higher incidence of adverse effects was associated with increasing age. Sixty-four percent of patients completed at least 6 months of isoniazid. Higher completion rates were associated with younger age, Hispanic ethnicity, and non-U.S. country of birth. Lower completion rates were associated with being homelessness, using excess alcohol, and having experienced an adverse effect. In summary, we conclude that in our clinic population isoniazid is a safe therapy for latent tuberculosis, but its effectiveness is limited by modest completion rates. PMID- 12746256 TI - Lack of a functional p21WAF1/CIP1 gene accelerates caspase-independent apoptosis induced by cisplatin in renal cells. AB - The lack of cyclin-dependent kinase inhibitor p21WAF1/CIP1 (p21) in mice increases renal proximal tubular cell death and enhances sensitivity to acute renal failure produced by the chemotherapeutic agent cisplatin. We used primary cultures of mouse renal proximal tubular cells (MPTC) grown in optimized culture conditions to investigate the cellular basis for increased apoptosis in p21 knockout mice. Cisplatin (15 microM) activated caspase-3 but not caspase-8 or caspase-9 and produced phosphatidylserine externalization, chromatin condensation, and nuclear fragmentation in wild-type [p21(+/+)] MPTC. Caspase-3 activation and apoptosis were accelerated in cisplatin-treated MPTC lacking p21 [p21(-/-) MPTC]. In contrast to p21(+/+) MPTC, cisplatin activated caspase-9 but not caspase-8 in p21(-/-) MPTC before caspase-3 activation. The caspase-3 inhibitor Asp-Glu-Val-Asp-fluoromethylketone (DEVD-fmk) inhibited caspase-3 activity but did not abolish apoptosis in p21(+/+) and p21(-/-) MPTC. General caspase inhibitor Z-Val-Ala-Asp(OCH3)-fluoromethylketone (ZVAD-fmk) inhibited caspase activity and decreased chromatin condensation by 51% in p21(-/-) but not in p21(+/+) MPTC. However, cisplatin-induced phosphatidylserine externalization was not inhibited by ZVAD-fmk in p21(-/-) MPTC. We conclude that 1) in the presence of p21, cisplatin activates caspase-3 through a mechanism independent of caspase-8 or caspase-9; 2) in the absence of p21, caspase-9 activation precedes caspase-3 activation; 3) the lack of p21 accelerates caspase-3 activation and cisplatin-induced MPTC apoptosis; and 4) MPTC apoptosis is caspase independent in the presence of p21 but partially dependent on caspases in the absence of p21. PMID- 12746257 TI - Effect of altered Na+ entry on expression of apical and basolateral transport proteins in A6 epithelia. AB - In several in vivo settings, prolonged alterations in the rate of apical Na+ entry into epithelial cells alter the ability of these cells to reabsorb Na+. We previously modeled this load dependence of transport in A6 cells by either decreasing Na+ entry via apical Na+ removal or amiloride or enhancing Na+ entry by chronic short-circuiting (Rokaw MD, Sarac E, Lechman E, West M, Angeski J, Johnson JP, and Zeidel ML. Am J Physiol Cell Physiol 270: C600-C607, 1996). Inhibition of Na+ entry by either method was associated with striking downregulation of transport rate as measured by short-circuit current (Isc), which recovered to basal levels of transport over a period of hours. Conversely, upregulation of Na+ entry by short-circuiting resulted in a sustained increase in transport rate that also returned to basal levels over a period of hours. The current studies were undertaken to determine whether these conditions were associated with alterations in either the whole cell content or apical membrane distribution of sodium channel (ENaC) subunits or on basolateral expression of either of the subunits of the Na+-K+-ATPase. We compared these effects to those achieved by chronic upregulation of Na+ transport by aldosterone. Whole cell levels of ENaC subunits were measured by immunoblot following 18-h inhibition of Na+ entry achieved by either tetramethylammonium replacement of Na+ or apical amiloride or after an 18-h increase in Na+ entry achieved by chronic short circuiting. None of these maneuvers significantly altered the whole cell content of any of the ENaC subunits compared with control cells. We then examined the effects of these maneuvers on apical membrane ENaC expression using domain specific biotinylation and immunoblot. Inhibition of Na+ entry by either method was associated with a profound decrease in apical membrane beta-ENaC without significant changes in apical membrane alpha-or gamma-ENaC amounts. Restoration of apical Na+ and/or removal of amiloride resulted in return of Isc to control levels over 2 h and coincided with return of apical beta-ENaC to control levels without change in apical alpha- or gamma-ENaC. Stimulation of Na+ transport by short-circuiting, in contrast, did not significantly alter apical membrane composition of any of the ENaC subunits. Basolateral expression of Na+-K+-ATPase was also measured by biotinylation and immunoblot and was unchanged under all conditions. Aldosterone increased basolateral expression of the alpha-subunit of Na+-K+-ATPase. These results suggest that chronic downregulation of transport is mediated, in part, by a selective decrease in apical membrane ENaC expression, consistent with our previous observations of noncoordinate regulation of ENaC expression under varying transport conditions in A6 cells. The chronic increase in the rate of Na+ entry is not associated with any of the changes in transporter density at either apical or basolateral membrane seen with aldosterone, suggesting that these two mechanisms of augmenting transport are completely distinct. PMID- 12746258 TI - Angiotensin blocks substance P release from renal sensory nerves by inhibiting PGE2-mediated activation of cAMP. AB - Activation of renal sensory nerves involves PGE2-mediated release of substance P (SP) via activation of the cAMP-PKA pathway. The PGE2-mediated SP release is suppressed by a low- and enhanced by a high-sodium (Na+) diet, suggesting an inhibitory effect of ANG. We now examined whether ANG II is present in the pelvic wall and inhibits PGE2-mediated SP release by blocking PGE2-mediated increases in cAMP. ANG II levels in renal pelvic tissue were 710 +/- 95 and 260 +/- 30 fmol/g tissue in rats fed a low- and high-Na+ diet, respectively. In a renal pelvic preparation from high-Na+-diet rats, 0.14 microM PGE2 produced an increase in SP release from 7 +/- 1 to 19 +/- 3 pg/min that was blocked by 15 nM ANG II. Treating pelvises with pertussis toxin (PTX) abolished the effects of ANG II. In pelvises from low-Na+ rats, neither basal nor bradykinin-mediated SP release was altered by PGE2. However, the bradykinin-mediated release of SP was enhanced by the permeable cAMP analog CPT-cAMP, from 4 +/- 1 to 11 +/- 2 pg/min, a response similar to that in normal-Na+-diet rats. In vivo, renal pelvic administration of PGE2 enhanced the afferent renal nerve activity (ARNA) response to bradykinin in normal- but not in low-Na+ diet rats. CPT-cAMP produced similar enhancement of the ARNA responses to bradykinin in normal- and low-Na+-diet rats, 1,670 +/- 490 and 1,760 +/- 400%.s (area under the curve of ARNA vs. time). Similarly, the ARNA responses to increases in renal pelvic pressure were similarly enhanced by CPT cAMP in normal- and low-Na+-diet rats. In conclusion, renal pelvic ANG II modulates the responsiveness of renal sensory nerves by suppressing PGE2-mediated activation of adenylyl cyclase via a PTX-sensitive mechanism. PMID- 12746259 TI - Immunolocalization of a microsomal prostaglandin E synthase in rabbit kidney. AB - PGE2, the major cyclooxygenase (COX) metabolite of arachidonic acid, is an important paracrine regulator of numerous tubular and vascular functions in the kidney. To date, COX activity has been considered the key step in prostaglandin synthesis and is well characterized. However, much less is known about the recently cloned microsomal PGE2 synthase (mPGES), the terminal enzyme of PGE2 synthesis, which converts COX-derived PGH2 to the biologically important PGE2. Present studies provide the detailed localization of mPGES protein in the rabbit kidney using immunohistochemistry. In the cortex, strong mPGES labeling was found in the macula densa (MD) and principal cells of the connecting segment and cortical collecting tubule but not in intercalated cells. The medulla was abundant in mPGES-positive structures, with heavy labeling in the collecting duct system. In descending thin limbs and renal medullary interstitial cells, mPGES expression was less intense, and it was below the limits of detection in the vasa recta. Expression of MD mPGES, similarly to COX-2, was greatly increased in response to low-salt diet and angiotensin I-converting enzyme inhibition by captopril. These findings suggest autocrine regulation of renal salt and water transport by PGE2 in descending thin limb and collecting tubule and a paracrine effect of PGE2 on the glomerular and medullary vasculature. Similar to other organs, mPGES in the kidney is an inducible enzyme and may be similarly regulated and acts in concert with COX-2. PMID- 12746261 TI - Tissue and cell imaging in situ: potential for applications in pathology and endoscopy. AB - Morphological sciences have recently experienced a significant technological breakthrough that offers new opportunities for cell and tissue imaging in situ but also raises new challenges to pathologists, who must adapt to a rapidly evolving environment. New partners, such as cell and molecular biologists, have provided pathologists with highly powerful tools for cell and subcellular imaging. They include: (a) the adaptation of techniques derived from molecular biology and cytogenetics, (b) the development of new microscopic tools, such as confocal microscopy, and (c) the emergence of new preparative techniques, such as microdissection or tissue arrays. However, recent technological progresses in various fields, from endoscopy to genomics, also raise new challenges to pathologists. Pathologists must therefore be prepared to redefine their area of expertise: this will be achieved through a continuous collaboration with all the partners involved in cell and tissue imaging and analysis but also by emphasising the importance of the informations provided by cell and tissue imaging in situ. PMID- 12746262 TI - Magnification endoscopy, high resolution endoscopy, and chromoscopy; towards a better optical diagnosis. AB - In the past few years, optical magnification endoscopy and chromoscopy have gained renewed interest in the West as a means for the early detection of minute lesions in patients with Barrett's oesophagus and in patients referred for colonic cancer screening. In Barrett's oesophagus, the vast majority of data on the use of chromoscopy deals with the application of methylene blue. Conventional videoendoscopy in combination with methylene blue staining improves the detection of Barrett's mucosa. A correlation has been shown between variation and intensity of staining and histologically verified stages of dysplasia or cancer. Magnification endoscopy and chromoscopy improve the detection of colonic non polypoid lesions associated with neoplasia and carcinoma. Pitt pattern analysis enables the distinction of non-neoplastic non-polypoid lesions (type I and II) from neoplastic type non-polypoid lesions (type III-V) with great accuracy. It is certain that "old fashioned" chromoscopy combined with advanced endoscopic technology carry a great diagnostic potential and should be further put to the test for use in daily clinical practice. PMID- 12746263 TI - Novel methods of enhanced endoscopic imaging. AB - Endoscopy has become an essential part of the practice of gastroenterology. Techniques exploiting previously unused properties of light have demonstrated the potential to enhance the ability to make clinical diagnoses without removing tissue as has been standard practice for decades. The term used for many of these techniques is "optical biopsy" and, although not yet widely available, enthusiasm for such techniques has grown as has research in their potential clinical utility. PMID- 12746264 TI - Virtual magnetic resonance colonography. AB - Colorectal cancer screening has vast potential. Beyond considerations for cost and diagnostic accuracy, the effectiveness of any colorectal screening strategy will be dependent on the degree of patient acceptance. Magnetic resonance (MR) colonography has been shown to be accurate regarding the detection of clinically relevant colonic polyps exceeding 10 mm in size, with reported sensitivity and specificity values exceeding 95%. To further increase patient acceptance, strategies for fecal tagging have recently been developed. By modulating the signal of fecal material to be identical to the signal characteristics of the enema applied to distend the colon, fecal tagging in conjunction with MR colonography obviates the need for bowel cleansing. The review will describe the techniques underlying MR colonography and describe early clinical experience with fecal tagging techniques. PMID- 12746266 TI - Improving endoscopic resolution and sampling: fluorescence techniques. AB - Gastrointestinal tumours are usually associated with a poor prognosis, as most are only diagnosed at an advanced stage. Every effort to improve intraluminal diagnosis in gastroenterology must therefore be aimed at diagnosing gastrointestinal tract tumours at the earliest stage possible. PMID- 12746265 TI - 18F-Fluoro-2-deoxyglucose positron emission tomography in the evaluation of gastrointestinal malignancies. AB - Positron emission tomography with (18)F-fluoro-2-deoxyglucose is an imaging technology that is demonstrating increasing utility in the evaluation of gastrointestinal malignancies. PMID- 12746267 TI - Non-invasive investigation of gastrointestinal functions with magnetic resonance imaging: towards an "ideal" investigation of gastrointestinal function. AB - Gastrointestinal (GI) function is complex and physiological measurements are subject to a variety of technical difficulties and practical limitations. The ideal technique would be non-invasive, widely available, convenient, and reliable and would not expose the subject to ionising radiation. It would permit direct assessment of GI function in the postprandial as well as the resting state, and be able to differentiate between food, secretion, and air in the lumen. GI structure and function are interdependent and the ideal technique would permit simultaneous assessment of these factors. Finally, the bowel operates as a functional whole and assessment of the GI tract proximal and distal to the area of interest is desirable. In this article the authors summarise the development and validation of magnetic resonance imaging techniques that overcome many of the deficiencies of existing methods, and have many characteristics of the "ideal" investigation of GI function. PMID- 12746269 TI - Wireless capsule endoscopy. AB - Gastroscopy, small bowel endoscopy, and colonoscopy are uncomfortable because they require comparatively large diameter flexible cables to be pushed into the bowel, which carry light by fibreoptic bundles, power, and video signals. Small bowel endoscopy is currently especially limited by problems of discomfort and failure to advance enteroscopes far into the small bowel. There is a clinical need for better methods to examine the small bowel especially in patients with recurrent gastrointestinal bleeding from this site. PMID- 12746268 TI - In vivo imaging with oligonucleotides for diagnosis and drug development. AB - Molecular imaging, the science that combines non-invasive in vivo imaging and molecular biology, has begun to use labelled oligonucleotides as radiotracers. Antisense oligonucleotides target gene expression at the RNA level, while aptamer oligonucleotides are designed to hit proteins of interest. Oligonucleotides for imaging cover a large range of applications, from the invention of new contrast agents for diagnosis to exquisite research tools for the development of new drugs. PMID- 12746271 TI - Minireview: overview of the renin-angiotensin system--an endocrine and paracrine system. AB - Since the discovery of renin as a pressor substance in 1898, the renin angiotensin (RAS) system has been extensively studied because it remains a prime candidate as a causative factor in the development and maintenance of hypertension. Indeed, some of the properties of the physiologically active component of the RAS, angiotensin II, include vasoconstriction, regulation of renal sodium and water absorption, and increasing thirst. Initially, its affect on blood pressure was thought to be mediated primarily through the classical endocrine pathway; that is, the generation of blood-borne angiotensin with actions in target tissues. More recently, however, it has become appreciated that a local autocrine or paracrine RAS may exist in a number of tissues, and that these may also play a significant role in regulating blood pressure. Some of the difficulties in studying tissue RAS stem from the limitations of pharmacology in not differentiating between RAS products made systemically from those synthesized locally. However, the development of transgenic animals with highly specific promoters to target the RAS to specific tissues provided important tools to dissect these systems. Thus, this minireview will discuss recent advances in understanding the relationship between endocrine and paracrine (tissue) RAS using transgenic models. PMID- 12746272 TI - Minireview: computer simulations of blood pressure regulation by the renin angiotensin system. AB - Gene targeting experiments in mice have been used by us and others to test whether quantitative changes in gene expression in the renin-angiotensin system affect blood pressure. Surprisingly, these studies showed that blood pressure does not change with mild quantitative changes in the expression of the angiotensin converting enzyme (ACE). Yet, ACE inhibitors are widely used for the treatment of hypertension. This apparent paradox motivated us to develop a simple computer simulation, which qualitatively reconciled the paradox. We have now improved the simulation by including blood pressure as an explicit variable and by adding the kallikrein-kinin system and feedback control of plasma renin via plasma angiotensin II levels. The new simulation now matches quantitative aspects of the experimental data and suggests that a decrease in bradykinin plays an important role in the increased risk of diabetic nephropathy associated with genetically determined higher levels of ACE activity. This emphasizes that the value of these types of simulation lies in the thoughts that they provoke rather than in their ability to replicate experimental data. PMID- 12746273 TI - Minireview: natriuretic peptides during development of the fetal heart and circulation. AB - Atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) are cardiac hormones, secreted by the atria and ventricles, respectively, in the normal adult heart. They participate in the regulation of blood pressure and body fluid homeostasis and modify growth and development of cardiovascular tissues and bone. Levels of ANP are higher in the fetal circulation than in adults, and fetal ventricles express higher levels of ANP and BNP than adult ventricles. The reappearance of ventricular ANP expression in adults is recognized as a marker of the induction of the embryonic gene program in ventricular hypertrophy. The natriuretic peptide system appears to be functional by midgestation, to respond to volume stimuli, and to regulate blood pressure and salt and water balance in the developing embryo. In addition, the natriuretic peptides may help regulate the blood supply to the fetus, acting as vasodilators in the placental vasculature. Peaks of ANP and BNP expression during gestation coincide with significant events in cardiac organogenesis, suggesting a role for ANP/BNP in the formation of the heart. In knockout mice lacking the natriuretic peptide receptor (NPR)-A gene (Npr1(-/-)), survival is reduced, with hearts enlarged at birth and possible cardiac developmental abnormalities. Surviving adult Npr1(-/-) mice have elevated blood pressure and marked cardiac hypertrophy and fibrosis, indicating that the ANP/BNP system is an important regulator of myocyte growth during development. PMID- 12746274 TI - Minireview: adiposity, inflammation, and atherogenesis. AB - Adipose tissue is a dynamic endocrine organ that secretes a number of factors that are increasingly recognized to contribute to systemic and vascular inflammation. Several of these factors, collectively referred to as adipokines, have now been shown regulate, directly or indirectly, a number of the processes that contribute to the development of atherosclerosis, including hypertension, endothelial dysfunction, insulin resistance, and vascular remodeling. Several adipokines are preferentially expressed in visceral adipose tissue, and the secretion of proinflammatory adipokines is elevated with increasing adiposity. Not surprisingly, approaches that reduce adipose tissue depots, including surgical fat removal, exercise, and reduced caloric intake, improve proinflammatory adipokine levels and reduce the severity of their resultant pathologies. Systemic adipokine levels can also be favorably altered by treatment with several of the existing drug classes used to treat insulin resistance, hypertension, and hypercholesterolemia. Greater understanding of adipokine regulation, however, should result in the design of improved treatment strategies to control disease states associated with increase adiposity, an important outcome in view of the growing worldwide epidemic of obesity. PMID- 12746275 TI - Minireview: lipid metabolism, metabolic diseases, and peroxisome proliferator activated receptors. AB - Lipid and carbohydrate homeostasis in higher organisms is under the control of an integrated system that has the capacity to rapidly respond to metabolic changes. The peroxisome proliferator-activated receptors (PPARs) are nuclear fatty acid receptors that have been implicated to play an important role in obesity-related metabolic diseases such as hyperlipidemia, insulin resistance, and coronary artery disease. The three PPAR subtypes, alpha, gamma, and delta, have distinct expression patterns and evolved to sense components of different lipoproteins and regulate lipid homeostasis based on the need of a specific tissue. Recent advances in identifying selective ligands in conjunction with microarray analyses and gene targeting studies have helped delineate the subtype-specific functions and the therapeutic potential of these receptors. PPARalpha potentiates fatty acid catabolism in the liver and is the molecular target of the lipid-lowering fibrates (e.g. fenofibrate and gemfibrozil), whereas PPARgamma is essential for adipocyte differentiation and mediates the activity of the insulin-sensitizing thiazolidinediones (e.g. rosiglitazone and pioglitazone). Recent evidence suggests that PPARdelta may be important in controlling triglyceride levels by sensing very low-density lipoprotein. Thus, uncovering the regulatory mechanisms and transcriptional targets of the PPARs will continue to provide insight into the pathogenesis of metabolic diseases and, at the same time, offer valuable information for rational drug design. PMID- 12746276 TI - Minireview: primary aldosteronism--changing concepts in diagnosis and treatment. AB - Primary aldosteronism affects 5-13% of patients with hypertension. Patients with hypertension and hypokalemia and most patients with treatment-resistant hypertension should undergo screening for primary aldosteronism with a plasma aldosterone concentration to plasma renin activity ratio. A high plasma aldosterone concentration to plasma renin activity ratio is a positive screening test result, a finding that warrants confirmatory testing. For those patients that want to pursue a surgical cure, the accurate distinction between the subtypes (unilateral vs. bilateral adrenal disease) of primary aldosteronism is a critical step. The subtype evaluation may require one or more tests, the first of which is imaging the adrenal glands with computed tomography, followed by selective use of adrenal venous sampling. Because of the deleterious cardiovascular effects of aldosterone, normalization of circulating aldosterone or aldosterone receptor blockade should be part of the management plan for all patients with primary aldosteronism. Unilateral laparoscopic adrenalectomy is an excellent treatment option for patients with unilateral aldosterone-producing adenoma. Bilateral idiopathic hyperaldosteronism should be treated medically. In addition, aldosterone-producing adenoma patients may be treated medically if the medical treatment includes mineralocorticoid receptor blockade. PMID- 12746277 TI - Endochondral bone formation in the heart: a possible mechanism of coronary calcification. AB - During the atherosclerotic process, calcification occurs and is associated with a high likelihood of adverse events. Coronary calcification has been perceived as a passive precipitation of mineral. Recently, calcification associated with atherosclerosis has been found to be the result of an organized, regulated process that is similar to the process of calcification in bone. Mineralization in skeletal tissue can form by endochondral ossification in which mesenchymal cells differentiate into chondroblasts and produce a cartilage matrix which then degenerates and is remodeled to form bone. In this study, hearts from oophorectomized, aged female Sprague Dawley rats were found to contain areas of cartilage. Micro-computerized tomography radiogrammetry provided quantitative images of the architecture and confirmed the calcified tissue. Histological analysis revealed staining for several markers consistent with cartilage and bone tissue: acid phosphatase and bone matrix proteins, osteocalcin, osteopontin, osteonectin, and bone sialoprotein. In addition, cartilage types II, X, and procollagen type I were present. The presence of chondrocytes in the aged rat heart provides insights into the process of calcification in coronary arteries. Many proteins associated with calcification in bone are present in the cartilage that is present in vascular tissue, suggesting that endochondral calcification is another possible mechanism by which calcification of vascular tissue may occur. PMID- 12746278 TI - The role of a conserved region of the second intracellular loop in AT1 angiotensin receptor activation and signaling. AB - The pleiotropic actions of angiotensin II are mediated by the primarily G(q) protein-coupled type 1 angiotensin (AT(1)) receptor. In this study a mutational analysis of the function of the conserved DRYXXV/IXXPL domain in the second intracellular loop of the rat AT(1A) receptor was performed in COS7 cells. Alanine substitution studies showed that single replacement of the highly conserved Asp(125) and Arg(126), but not Tyr(127), moderately impaired angiotensin II-induced inositol phosphate signaling. However, concomitant substitution of both Asp(125) and Arg(126) caused marked reduction of both inositol phosphate signaling and receptor internalization. Alanine scanning of the adjacent residues showed that substitution of Ile(130), His(132), and Pro(133) reduced agonist-induced inositol phosphate signal generation, whereas mutations of Met(134) also impaired receptor internalization. Expression of the D125A mutant AT(1A) receptor in COS7 cells endowed the receptor with moderate constitutive activity, as indicated by its enhanced basal Elk1 promoter activity and inositol phosphate response to partial agonists. Angiotensin II-induced stimulation of the Elk1 promoter showed parallel impairment with inositol phosphate signal generation in receptors containing mutations in this region of the AT(1A) receptor. These data confirm that Ca(2+) signal generation is required for the nuclear effects of angiotensin II-induced ERK activation. They are also consistent with the role of the conserved DRY sequence of the AT(1A) receptor in receptor activation, and of Asp(125) in constraining the receptor in its inactive conformation. Furthermore, in the cytoplasmic helical extension of the third helix, an apolar surface that includes Ile(130) and Met(134) appears to have a direct role in G protein coupling. PMID- 12746279 TI - The angiotensin II type 1 receptor mediates renal interstitial content of tumor necrosis factor-alpha in diabetic rats. AB - A unique microdialysis technique was used to demonstrate that increased levels of angiotensin II (Ang II) and consequent stimulation of the Ang II type 1 (AT(1)) receptor increase the renal content of TNF-alpha in diabetes. Recovery of Ang II and TNF-alpha in renal interstitial fluid (RIF) was measured in conscious rats before and weekly for 12 wk after induction of diabetes with streptozocin and in response to oral valsartan (10 mg/kg.d). Recovery of Ang II in RIF was significantly higher in diabetic rats than in nondiabetic rats. In diabetic rats, RIF recovery of TNF-alpha increased by approximately 67% over baseline, whereas it was unchanged in nondiabetic rats. AT(1) receptor blockade with valsartan prevented the increase in TNF-alpha in the diabetic group. This study shows that diabetes is associated with an increase in the vasoconstrictive hormone Ang II and the inflammatory cytokine TNF-alpha, both of which play a role in accelerating renal function decline in diabetic nephropathy. The study also confirms that valsartan reduces intrarenal level of TNF-alpha by acting on Ang II at the AT(1) receptor level. This finding of a potential antiinflammatory effect for valsartan is new and in addition to its known antihypertensive effects. PMID- 12746280 TI - Urocortin-related peptides increase interleukin-6 output via cyclic adenosine 5' monophosphate-dependent pathways in A7r5 aortic smooth muscle cells. AB - Corticotropin-releasing factor receptor type 2beta, expressed in the rodent cardiovascular system, is a member of the G protein-coupled receptor family. This receptor is coupled positively to adenylate cyclase and is bound preferentially by the urocortin (Ucn)-related peptides (Uncs): Ucn, Ucn II, and Ucn III. In the present study, we investigated the effects of Ucns on IL-6 levels in A7r5 aortic smooth muscle cells. In this cell line, both Ucn and Ucn II induced accumulation of intracellular cAMP via corticotropin-releasing factor receptor type 2beta and also caused a significant increase in IL-6 output levels. The adenylate cyclase inhibitor, MDL-12330A, inhibited this Ucn- or Ucn II-induced increase in IL-6 levels. Although H89 (10 micro M), a protein kinase A inhibitor, had no effect on the increase in IL-6 concentration, bisindolylmaleimide I (10 nM), a protein kinase C inhibitor, was found to significantly inhibit IL-6 output levels. Blockade of Ucn- or Ucn II-induced increases in IL-6 levels by SB203580 (100 nM), a p38 MAPK inhibitor, suggested that the p38 MAPK pathway was involved in this regulation. The cAMP-mediated increase in IL-6 levels was suppressed synergistically by both bisindolylmaleimide I and SB203580. These findings demonstrate that both protein kinase C and p38 MAPK signaling cascades are involved downstream of the Ucns-cAMP pathway in A7r5 aortic smooth muscle cells. PMID- 12746282 TI - The endocrinology and physiology of parturition: understanding the process through mining genome databases. PMID- 12746281 TI - Cardiac function in mice lacking the glucagon-like peptide-1 receptor. AB - Glucagon-like peptide-1 (GLP-1) acts via its G protein-coupled receptor (GLP-1R) to regulate blood glucose. Although the GLP-1R is widely expressed in peripheral tissues, including the heart, and exogenous GLP-1 administration increases heart rate and blood pressure in rodents, the physiological importance of GLP-1R action in the cardiovascular system remains unclear. We now show that 2-month-old mice with genetic deletion of the GLP-1R (GLP-1R(-/-)) exhibit reduced resting heart rate and elevated left ventricular (LV) end diastolic pressure compared with CD-1 wild-type controls. At the age of 5 months, echocardiography and histology demonstrate increased LV thickness in GLP-1R(-/-) mice. Although baseline hemodynamic parameters of GLP-1R(-/-) did not differ significantly from those of wild type, GLP-1R(-/-) mice displayed impaired LV contractility and diastolic function after insulin administration. The defective cardiovascular response to insulin was not attributable to a generalized defect in the stress response, because GLP-1R(-/-) mice responded appropriately to insulin with increased c-fos expression in the hypothalamus and increased circulating levels of glucagon and epinephrine. Furthermore, LV contractility after exogenous epinephrine infusion was also reduced in GLP-1R(-/-) mice. These findings provide new evidence implicating an essential role for GLP-1R in the control of murine cardiac structure and function in vivo. PMID- 12746283 TI - Gene expression profiling of rat uterus at different stages of parturition. AB - A fuller understanding of the process of parturition is needed in view of the current lack of efficient treatment for preterm labor. Using DNA microarrays, we have analyzed patterns of uterine gene expression at d 0 and 20 of pregnancy, at term in labor or not in labor, and at 1 d post partum. Of the 8740 genes analyzed, 562 genes undergoing significant changes were grouped into 5 distinct clusters, each containing many genes not previously known to be involved with uterine functions. Cluster 1 genes were up-regulated at labor and encompassed immune defense and immediate early response genes, including transcription factors NGFI-B/nurr77 and estrogen-responsive gene 1. Cluster 3 genes were acutely suppressed at labor and included extracellular matrix products and genes related to hormonal signaling, implying novel intrauterine mechanisms regulating intracellular cyclic GMP and local steroid hormone concentrations. At labor, more genes were suppressed than activated, indicating that, for the process of labor induction, gene suppression is at least equally important as the more extensively studied processes of gene activation. The study also points to the existence of novel uterine signaling pathways, including Wnt/frizzled and receptor activator of nuclear factor-kappaB (RANK) and its ligand, as well as the involvement of novel signaling molecules such as estrogen-responsive gene 1, decay-accelerating factor 1, and ebnerin. The present results provide the basis for further studies that will enlarge our knowledge of the mechanisms underlying labor and parturition under physiological and pathophysiological conditions. PMID- 12746284 TI - Paracrine prolactin may cause prostatic problems. PMID- 12746285 TI - Prostate hyperplasia in a transgenic mouse with prostate-specific expression of prolactin. AB - Prolactin (PRL) is one of several polypeptide factors known to exert trophic effects on the prostate. We have previously reported a dramatic prostate enlargement with concurrent chronic hyperprolactinemia and elevated serum androgen levels in a PRL transgenic mouse (Mt-PRL) with ubiquitous expression of the transgene. To address the role of local PRL action in the prostate, a new transgenic mouse model (Pb-PRL) was generated using the prostate-specific rat probasin (Pb) minimal promoter to drive expression of the rat PRL gene. Pb-PRL transgenic males developed a significant enlargement of both the dorsolateral and ventral prostate lobes evident from 10 wk of age and increasing with age. Expression of the transgene was restricted to the prostate and detected from 4 wk of age. Low levels of transgenic rat PRL were detectable in the serum of adult Pb PRL animals. Serum androgen levels were normal. The Pb-PRL prostate displayed significant stromal hyperplasia, ductal dilation, and focal areas of epithelial dysplasia. Quantitative analysis of prostatic tissue cellularity demonstrated a marked increase in the stromal to epithelial ratio in all lobes of Mt-PRL and Pb PRL transgenic prostates compared with controls. Microdissections demonstrated an increased ductal morphogenesis in dorsolateral and ventral prostate lobes of Mt PRL prostate vs. Pb-PRL and controls. In conclusion, this study indicates the ability of PRL to promote, directly or indirectly, ductal morphogenesis in the developing prostate and further to induce abnormal growth primarily of the stroma in the adult gland in a setting of normal androgen levels. PMID- 12746286 TI - Gene expression, secretion, and autocrine action of C-type natriuretic peptide in cultured adult rat cardiac fibroblasts. AB - C-type natriuretic peptide (CNP), the third member of the natriuretic peptide family, is known to be synthesized in the central nervous system and vascular endothelial cells, in contrast to atrial natriuretic peptide and brain natriuretic peptide. However, there have been no studies concerning CNP production in cultured cardiac cells. Here, we examined the production and the local effect of CNP in cultured ventricular cells. Under serum-free conditions, adult rat cardiac fibroblasts secreted immunoreactive CNP time dependently. TGF beta1, basic fibroblast growth factor, and endothelin-1 significantly stimulated CNP secretion. Northern blot analysis detected significant expressions of CNP and its specific receptor (guanylyl cyclase-B) mRNA in cardiac fibroblasts. CNP stimulated intracellular cGMP production in fibroblasts more intensely than atrial and brain natriuretic peptides. CNP inhibited both DNA and collagen syntheses of cardiac fibroblasts, and these inhibitory effects by CNP were stronger than by atrial and brain natriuretic peptides. The inhibition by CNP of DNA and collagen syntheses was reproduced by a cGMP analog, 8-bromo cGMP. The present findings demonstrate that CNP is synthesized in and secreted from cardiac fibroblasts and suggest that CNP has a suppressive effect on fibroblast proliferation and extracellular matrix production, probably via the guanylyl cyclase-B-mediated cGMP-dependent process. CNP produced by cardiac fibroblasts may play a role as an autocrine regulator against excessive cardiac fibrosis. PMID- 12746287 TI - High levels of intrauterine corticotropin-releasing hormone, urocortin, tryptase, and interleukin-8 in spontaneous abortions. AB - Stress induces CRH secretion that activates hypothalamic-pituitary-adrenal axis and is also abortogenic. In addition to hypothalamus, CRH and its analog urocortin (Ucn) are also secreted locally outside the brain where they activate mast cells leading to inflammation; however, the level of CRH and Ucn or mast cell mediators has not been examined in products of conception (POC). CRH and Ucn were measured by enzyme immunoassay, tryptase by fluoroenzyme immunoassay, and IL 8 by ELISA in POC of 7-9 wk gestation from Caucasian women; they were divided into group I with elective abortions (n = 4), group II with one spontaneous abortion (n = 12), and group III with at least two spontaneous abortions (n = 7). CRH, Ucn, tryptase, and IL-8 levels were higher (P < 0.05) in group III (8683 +/- 1201 pg/g, 7961 +/- 1499 pg/g, 1553 +/- 572 ng/g, and 8317 +/- 1874 pg/g, respectively) than group II (2561 +/- 314 pg/g, 2349 +/- 394 pg/g, 403 +/- 97 ng/g, and 3199 +/- 449 pg/g, respectively) and group I (163 +/- 162 pg/g, 328 +/- 327 pg/g, 72 +/- 31 ng/g, and 3681 +/- 931 pg/g, respectively). Immunostaining of POC showed significantly more tryptase in group III women. High POC levels of CRH and Ucn under stress in habitual spontaneous abortions may activate uterine mast cells to secrete abortogenic tryptase and IL-8. PMID- 12746288 TI - Female oxytocin-deficient mice display enhanced anxiety-related behavior. AB - Previous studies have suggested that oxytocin (OT) may be anxiolytic in female laboratory rats and mice. The elevated plus-maze was used to compare anxiety related behaviors of OT-deficient (OT-/-) and wild-type (OT+/+) mice. Female OT-/ mice displayed increased anxiety-related behavior compared with OT+/+ mice. The percentage of entries (P < 0.0002) and time spent (P < 0.003) in the open arms was less in female OT-/- than OT+/+ mice. Administration of synthetic OT, 2 ng by intracerebroventricular (icv) injection to female OT-/- mice, increased the percentage of entries (P < 0.003) and time spent (P < 0.004) in the open arms compared with artificial cerebrospinal fluid female OT-/- mice. Administration of an OT receptor antagonist (Atosiban, d[Dtyr(Et)(2), Thr(4)]ornithine vasotocin) 100 ng icv, to female OT+/+ mice increased anxiety-related behavior by decreasing the percentage of entries (P < 0.01) and time spent (P < 0.04) in the open arms compared with artificial cerebrospinal fluid-treated controls. Central infusion of an OT receptor antagonist, 100 ng icv, before administration of synthetic OT, 2 ng icv, in female OT-/- mice blocked the anxiolytic affect of OT. In contrast, male OT-/- mice displayed decreased anxiety-related behavior compared with male OT+/+ mice. The percentage of entries (P < 0.007) and time spent (P < 0.004) in the open arms was greater in male OT-/- vs. OT+/+ mice. Our findings indicate that OT pathways play a role in modulating anxiety in female mice of the C57BL/6 background, and the effect is mediated by the OT receptor. PMID- 12746289 TI - Identification of 3,5-diiodo-L-thyronine-binding proteins in rat liver cytosol by photoaffinity labeling. AB - In this study, we obtained evidence for the presence of cytosolic-binding proteins for 3,5-diiodo-L-thyronine (3,5-T(2)). UV irradiation of rat liver cytosol with [(125)I]3,5-T(2) resulted in specific covalent attachment of (125)I to three polypeptides with apparent molecular masses of 86, 66, and 38 kDa. The photoaffinity labeling of all three proteins was strongly inhibited (by about 90%) when the reaction was carried out in the presence of a 10-fold excess of unlabeled 3,5-T(2) or T(3). However, whereas inhibition by 3,5-T(2) was nicotinamide adenine dinucleotide phosphate reduced (NADPH) independent, T(3) inhibited only in the presence of NADPH. The 38-kDa protein, which showed the greatest affinity for 3,5-T(2), was partially purified by preparative fast performance liquid chromatography. Its binding activity was optimal at pH 7.4, stable between 0 and 37 C, and already maximal after 5-10 min of incubation. The finding that a 38-kDa cytosolic-binding protein binds 3,5-T(2) in the absence of NADPH, but T(3) only in a NADPH-dependent manner, suggests that it may serve to regulate intracellular T(3)/3,5-T(2) translocation in a way that depends on the nicotinamide adenine dinucleotide phosphate/NADPH ratio. PMID- 12746290 TI - Activation of cell adhesion kinase beta by mechanical stretch in vascular smooth muscle cells. AB - We have studied whether activation of cell adhesion kinase beta (CAKbeta) is involved in stretch-induced signaling pathway in cultured rat vascular smooth muscle cells. Cyclic stretch (1 Hz) induced a rapid (within 1 min) phosphorylation of CAKbeta, whose effect was time and strength dependent. Both Ca(2+) and Na(+) ionophores (A23187 and monensin) stimulated phosphorylation of CAKbeta in a similar fashion to mechanical stretch. The stretch-induced phosphorylation of CAKbeta was inhibited completely by an intracellular Ca(2+) chelator [1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis(acetoxymethyl ester)] and largely by gadolinium, but only partially by an extracellular Ca(2+) chelator (EGTA). An angiotensin type 1 receptor antagonist (CV11974) abolished the phosphorylation of CAKbeta stimulated by angiotensin II, but not by mechanical stretch. Mechanical stretch rapidly (within 1 min) increased the association of CAKbeta with c-Src, but not pp125(focal adhesion kinase). Stretch-induced phosphorylation of ERK1/2 was inhibited by EGTA and an inhibitor of the Src kinase family [4-amino-5-(4-chlorophenyl)-7-(t butyl)pyrazolo[3,4-d]pyrimidine], but not by cytochalasin D, to disrupt actin polymerization. 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine or cytochalasin D did not affect stretch-induced phosphorylation of CAKbeta. These data suggest that mechanical stretch stimulates activation of CAKbeta, followed by its association with c-Src, which requires ion influx mainly via stretch-activated nonselective ion channels, thereby leading to activation of the p21(Ras)/ERK1/2 cascade in vascular smooth muscle cells. PMID- 12746291 TI - Cytochrome P450 genes are differentially expressed in female and male hepatocyte retinoid X receptor alpha-deficient mice. AB - To study the functional role of retinoid X receptor alpha (RXRalpha) in hepatocytes, hepatocyte RXRalpha-deficient mice have been established. Characterization has been performed on male mice. In this paper, we show that the expression of CYP450 genes is differentially expressed in male and female hepatocyte RXRalpha-deficient mice; male mice have reduced expression of cytochrome P450 (CYP) CYP4A, CYP3A, and CYP2B mRNAs, but females do not exhibit such phenotypes. To examine the hormonal effects on this sexual dimorphic phenotype, male and female mice were subjected to 17beta-estradiol and 5alpha dihydrotestosterone (DHT) treatment, respectively, and then the expression of the CYP450 genes was studied. Estradiol had no effect on protecting the hepatocyte RXRalpha-deficient mice from reduced expression of the CYP450 genes. In contrast, DHT induced hepatocyte RXRalpha-deficient female mice, but not wild-type female mice, to have the reduced expression of CYP450 mRNAs. In addition, castration prevented the mutant male mice from exhibiting reduced expression of CYP450 mRNAs. wild-type and mutant mouse livers from both genders express androgen receptors (ARs). By transient transfection, DHT-AR could inhibit RXRalpha mediated transcription. Furthermore, by transfection and coimmunoprecipitation, RXR can interact with AR in vivo. These data suggest that testosterone has a negative impact on retinoid signaling when the level of RXRalpha is low, which may in turn reduce the expression of the CYP450 genes. PMID- 12746292 TI - Insulin-like growth factor I (IGF-I) and IGF binding protein-1 modulate prostate cancer cell growth and apoptosis: possible mediators for the effects of diet and exercise on cancer cell survival. AB - Previous studies showed that serum from men consuming a low fat diet and undergoing exercise intervention (DE) reduced LNCaP cell growth and induced apoptosis in vitro. DE also decreased serum IGF-I and increased serum IGF binding protein-1 (IGFBP-1). The present study evaluates the effects of IGF-I and IGFBP-1 on growth and apoptosis of prostate cancer cells in vitro. When IGF-I was added to the post-DE serum, the reduction in LNCaP cell growth and the induction of apoptosis in medium containing post-DE serum alone were reversed. When IGFBP-1 was added to the pre-DE serum samples, LNCaP cell growth was reduced, and apoptosis was induced. IGF-I, long-R(3)-IGF-I (only binds IGF-I receptor), AL(31)Leu(60)-IGF-I (only binds IGFBPs), antihuman IGF-I receptor antibodies, and IGFBP-1 were then added to LNCaP cultures to determine the independent effects of IGF-I and IGFBP-1 on cell growth. Collectively, the results using these agents show that IGF-I and IGFBP-1 exert opposing effects on LNCaP cell growth and apoptosis, and IGFBP-1 acts mainly through an IGF-dependent mechanism. DE results in a decrease in serum IGF-I with increased IGFBP-1 in vivo that is associated with apoptosis and reduced LNCaP and LAPC-4 prostate cancer cell growth in vitro. PMID- 12746293 TI - Estrogen receptor/Sp1 complexes are required for induction of cad gene expression by 17beta-estradiol in breast cancer cells. AB - The cad gene is trifunctional and expresses carbamoylphosphate synthetase/aspartate carbamyltransferase/dihydroorotase, which are required for pyrimidine biosynthesis. Cad gene activities are induced in MCF-7 human breast cancer cells, and treatment of MCF-7 or ZR-75 cells with 10 nM 17beta-estradiol (E2) resulted in a 3- to 5-fold increase in cad mRNA levels in both cell lines. The mechanism of hormone-induced cad gene expression was further investigated using constructs containing the growth-responsive -90 to +115 (pCAD1) region of the cad gene promoter. E2 induced reporter gene (luciferase) activity in MCF-7 and ZR-75 cells transfected with pCAD1, which contains three upstream GC-rich and two downstream E-box motifs. Deletion and mutation analysis of the cad gene promoter demonstrated that only the GC boxes that bind Sp1 protein were required for E2 responsiveness. Results of electrophoretic mobility shift and chromatin immunoprecipitation assays show that both Sp1 and estrogen receptor alpha interact with the GC-rich region of the cad gene promoter. Moreover, in transactivation assays with pCAD1, hormone-induced transactivation was inhibited by cotransfection with dominant-negative Sp1 expression plasmid and small inhibitory RNA for Sp1, which silences Sp1 expression in the cells. These results demonstrate that, in common with many other genes involved in E2-induced cell proliferation, the cad gene is also regulated by a nonclassical ERalpha/Sp1 mediated pathway. PMID- 12746294 TI - Molecular cloning, pharmacological characterization, and brain mapping of the melanocortin 4 receptor in the goldfish: involvement in the control of food intake. AB - We report cloning, pharmacological characterization, tissue distribution, detailed brain mapping, and role in control of food intake of melanocortin 4 receptor in goldfish (gMC4R). The gMC4R protein has 68% identity with the human ortholog and is conserved in important functional domains. Pharmacological profiling showed similar affinities and potency order to hMC4R for MSH peptides, whereas MTII and HS024 were identified as high-affinity agonist and antagonist analogs, respectively. The gMC4R-mRNA was found in brain and some peripheral tissues including the ovary, gill, and spleen. Detailed MC4R-mRNA mapping showed expression in main neuroendocrine and food intake-controlling areas. High expression levels were found in the telencephalon, preoptic area, ventral thalamus, tuberal hypothalamus, and hypothalamic inferior lobe. By RT-PCR, low levels were also detected in the cerebellum, medulla, and spinal cord. Intracerebroventricular MTII administration inhibited food intake in 24-h fasted animals in a dose-dependent manner, whereas HS024 stimulated food intake in fed animals, suggesting that melanocortins exert a tonic inhibitory effect on food intake, which is mediated through central MC4R signaling. The conserved central expression pattern and physiological role in regulation of food intake for the MC4R suggests that neuronal pathways of the melanocortin system may be important for regulation of energy homeostasis in most vertebrates. PMID- 12746296 TI - Follicle-stimulating hormone suppresses cytosolic 3,5,3'-triiodothyronine-binding protein messenger ribonucleic acid expression in rat granulosa cells. AB - FSH plays crucial roles in differentiation of granulosa cells and development of follicles. Considering the broad scope of FSH effects, a large number of genes are likely responsive to the hormone. However, only a limited number of genes have been identified as FSH-regulated genes, particularly during the preantral stage. In an attempt to better define genes involved in follicular development, we examined primary granulosa cell cultures, an undifferentiated rat ovarian granulosa cell line and rat ovaries, using differential display, quantitative RT PCR, Northern blot analysis, and in situ hybridization. We report, for the first time, that nicotinamide adenine dinucleotide phosphate-dependent cytosolic T(3) binding protein mRNA is expressed in the ovary, particularly in the granulosa cell layer of preantral and early antral follicles, but not in large preovulatory follicles. Its expression markedly declines in response to FSH, which is dependent on the period of the exposure. This FSH-responsive down-regulation is dependent on granulosa cell differentiation and follicular development. FSH down regulates the mRNA via the adenylyl cyclase/cAMP pathway, and the down-regulation requires de novo synthesis of a regulatory protein(s). The cytosolic T(3)-binding protein may play a significant role in the regulation of steroidogenesis and follicular development in the mammalian ovary. PMID- 12746295 TI - Estradiol exacerbates hippocampal damage in a model of preterm infant brain injury. AB - We have developed a model for prenatal hypoxia-ischemia in which muscimol, a selective gamma-aminobutyric acid A (GABA(A)) receptor agonist, administered to newborn rats, induces hippocampal damage. In the neonatal rat brain, activation of GABA(A) receptors leads to membrane depolarization and neuronal excitation. Because of our previous detection of sex differences in this model and the considerable interest in the neuroprotective effects of estradiol in the adult brain, we now investigate the effect of pretreatment with high physiological levels of estradiol in our model of prenatal hypoxia-ischemia. We used unbiased stereology to assess neuron number in the hippocampal formation of control, muscimol-treated, and estradiol- plus muscimol-treated animals. Muscimol decreased neuron number in the hippocampus, with damage exacerbated by pretreatment with estradiol. A hippocampal culture paradigm was developed to mirror the in vivo investigation. We observed elevated cytotoxicity (using the lactate dehydrogenase assay) by 48 h after treatment with estradiol plus muscimol, but decreased cytotoxicity between 2 and 24 h after treatment. To determine whether the actions of estradiol on muscimol-induced damage were via the estrogen receptor, hippocampal cultures were pretreated with ICI 182,780, a selective estrogen receptor antagonist. Treatment with ICI 182,780 blocked the potentiating effect of estradiol on the late period of cytotoxicity, but had no effect on the protective actions of estradiol during the early period of cytotoxicity. There appears to be a biphasic action of estradiol in our model of neonatal brain injury that involves early nongenomic, nonreceptor-mediated protection, followed by late deleterious receptor-mediated effects. PMID- 12746297 TI - Microarray and suppression subtractive hybridization analyses of gene expression in pheochromocytoma cells reveal pleiotropic effects of pituitary adenylate cyclase-activating polypeptide on cell proliferation, survival, and adhesion. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) exerts trophic effects on several neuronal, neuroendocrine, and endocrine cells. To gain insight into the pattern of the transcriptional modifications induced by PACAP during cell differentiation, we studied the effects of this neuropeptide on rat pheochromocytoma PC12 cells. We first analyzed the transcriptome of PC12 cells in comparison to that of terminally differentiated rat adrenomedullary chromaffin cells, using a high-density microarray, to identify genes associated with the proliferative phenotype that are possible targets of PACAP during differentiation of sympathoadrenal normal and tumoral cells. We then studied global gene expression in PC12 cells after 48 h of exposure to PACAP, using both cDNA microarray and suppression subtractive hybridization technologies. These complementary approaches resulted in the identification of 75 up-regulated and 70 down-regulated genes in PACAP-treated PC12 cells. Among the genes whose expression is modified in differentiated cells, a vast majority are involved in cell proliferation, survival, and adhesion/motility. Expression changes of most of these genes have been associated with progression of several neoplasms. A kinetic study of the effects of PACAP on some of the identified genes showed that the neuropeptide likely exerts early as well as late actions to achieve the gene expression program necessary for cell differentiation. In conclusion, the results of the present study underscore the pleiotropic role of PACAP in cell differentiation and provide important information on novel targets that could mediate the effects of this neuropeptide in normal and tumoral neuroendocrine cells. PMID- 12746298 TI - The FK506-binding immunophilin FKBP51 is transcriptionally regulated by progestin and attenuates progestin responsiveness. AB - FKBP51 and FKBP52 are large molecular weight FK506-binding immunophilins that have diverse biochemical functions. Best studied is the role that they play as components of steroid hormone receptors. Differential display and gene array screens have identified FKBP51 as a progestin-inducible gene. Here we demonstrate progestin enhancement of FKBP51 mRNA and protein in T-47D cells. FKBP51 mRNA and protein levels were increased 3-fold by 20 nM R5020. Induction of FKBP51 mRNA was unaffected by 1 micro g/ml cycloheximide but was blocked by the progestin receptor (PR) antagonist RU486 (1 micro M). Reporter plasmids containing 3.4 kb and 427 bp of 5'-flanking sequences of the human FKBP51 protein gene (FKBP5) exhibited regulation by progestin in T-47D cells. A construct containing 19 bp of upstream sequence demonstrated diminished basal activity and no stimulation by R5020. To test whether elevated FKBP51 affects progestin responsiveness, HepG2 cells were transfected with human FKBP51, PR, and mouse mammary tumor virus luciferase plasmids, and treated with R5020 (0.03-10 nM). Expression of FKBP51 increased the EC(50) for PR transactivation by 3.2-fold. Expression of FKBP51 from squirrel monkey, a New World primate with naturally occurring progestin resistance, increased the EC(50) more dramatically (11.7-fold vs. control). Expression of FKBP51 bearing a double-point mutation in the tetratricopeptide repeat domain had no effect on PR transactivation. These results suggest that increased expression of FKBP51 by progestin may attenuate progestin responsiveness in hormone-conditioned cells. Furthermore, overexpression of FKBP51 in the squirrel monkey may be a contributing cause of progesterone resistance in this species. PMID- 12746299 TI - Glucosamine induces resistance to insulin-like growth factor I (IGF-I) and insulin in Hep G2 cell cultures: biological significance of IGF-I/insulin hybrid receptors. AB - IGF-I stimulates insulin-like actions directly through its receptor, and it also enhances sensitivity to insulin-mediated effects in vivo. These studies were undertaken to analyze the role of IGF-I, insulin, and insulin/IGF-I hybrid receptors (HRs) in mediating IGF-I and insulin signaling in cells that had been made insulin-resistant by treatment with glucosamine. Human HepG2 cells, which express IGF-I receptors, insulin receptors (IRs), and IGF-I/insulin HRs, were exposed to 20 mM glucosamine; and the effects of IGF-I and insulin in stimulating glycogen synthesis were determined. An overnight exposure to glucosamine markedly attenuated the effects of insulin and IGF-I in stimulating glycogen synthesis. To determine which receptors were mediating this effect, the ability of insulin and IGF-I to stimulate phosphorylation of their respective receptors was analyzed. An 18-h exposure to glucosamine (20 mM) caused a 75% reduction in the ability of IGF I to phosphorylate its receptor but no change in receptor abundance. Glucosamine also caused a major reduction in insulin-stimulated receptor phosphorylation, although, unlike IGF-I, there was also a 50% reduction in IR abundance. Exposure to glucosamine also resulted in a reduction in the ability of IGF-I or insulin to stimulate phosphorylation of insulin IGF-I/HRs. The combination of insulin plus IGF-I was a more potent stimulus of HR phosphorylation than either agent alone, and this combination was also more potent in partially reversing the inhibitory effect of glucosamine. Taken together, these findings indicate that glucosamine induces a loss of sensitivity to stimulation of insulin, IGF-I, or HR tyrosine kinase activity by insulin or IGF-I. Although insulin is able to partially reverse the effect of glucosamine on IR phosphorylation, it has a very minimal effect on glucosamine-induced inhibition of HR phosphorylation. However, the combination of IGF-I and insulin induces a major increase in HR phosphorylation, even in the presence of glucosamine, suggesting that it is improving the sensitivity of the HR to insulin activation. PMID- 12746300 TI - Role of corticotropin-releasing factor receptors type 1 and 2 in modulating the rat adrenocorticotropin response to stressors. AB - We investigated the contribution of corticotropin-releasing factor (CRF) receptors types 1 and 2 (CRF(1) and CRF(2)) in mediating the ACTH response to shock, alcohol injection, or endotoxemia in the rat. Peptidic (Astressin B and Astressin(2)-B) and nonpeptidic (NBI 30775) CRF antagonists were injected iv before the stressors at doses previously shown to be effective in blocking the corresponding receptors. Because NBI 30775, which specifically blocks CRF(1), penetrates the brain following systemic injection, we also compared its effect with that of Astressin B, which primarily, though not exclusively, targets CRF(1) but does not cross the blood-brain barrier. Shocks, alcohol (4.5 g/kg, intragastrically) or lipopolysaccharide (LPS, 1 micro g/kg, iv) all significantly released ACTH. Astressin B or NBI 30775 markedly decreased the effect of shocks or alcohol and also interfered, though less significantly so, with the influence of LPS. In contrast, specific blockade of CRF(2) with Astressin(2)-B, although not significantly altering the overall ACTH response to shocks, alcohol, or LPS, slightly enhanced ACTH levels during the early phase of some of these responses. Interestingly, combined administration of NBI 30775 and Astressin(2)-B decreased ACTH levels more than NBI 30775 alone, although this difference did not reach statistical significance. Finally, blockade of CRF(1) and/or CRF(2) augmented LPS induced TNF-alpha and IL-6 release. Collectively, there results confirm the critical role played by CRF(1) in mediating the ACTH response to shocks, alcohol and LPS, whereas the influence of CRF(2) remains subtle. Finally, we showed that peripheral endogenous CRF restrains the ability of LPS to release cytokines. PMID- 12746301 TI - In vitro inhibition of insulin-degrading enzyme by long-chain fatty acids and their coenzyme A thioesters. AB - Insulin-degrading enzyme is responsible for initiating insulin degradation in cells, but little is known about the factors controlling its activity. Because obesity and high levels of free fatty acids decrease insulin clearance, we examined the effect of some common free fatty acids and their acyl-coenzyme A thioesters on insulin-degrading enzyme partially purified from the livers of male Sprague Dawley rats. Octanoic acid (C8:0) had no effect on activity. Long-chain free fatty acids (C16-C20) inhibited between 50% and 90% of the insulin degradation with IC(50) values in the range of 10-50 micro M. In general, the corresponding acyl-coenzyme A thioesters had lower IC(50) values and were slightly more efficacious. (125)I-insulin cross-linking studies showed free fatty acids did not inhibit hormone binding to insulin-degrading enzyme. Kinetic analysis showed a noncompetitive type of inhibition. Furthermore, fatty acids eliminated the ability of insulin to inhibit the proteasome. These results suggest that when intracellular long-chain fatty acid concentrations are elevated, they may act directly on insulin-degrading enzyme to decrease insulin metabolism and alter insulin action in intact cells. This mechanism may contribute to the hyperinsulinemia and insulin resistance seen with elevated fatty acids and obesity. PMID- 12746303 TI - Insulin-like growth factor-II bound to vitronectin enhances MCF-7 breast cancer cell migration. AB - We have previously reported that IGF-II binds the extracellular matrix protein vitronectin (VN) with an affinity similar to that for the type-1 IGF receptor (IGF-1R). In view of this finding, and given the cited role of VN in cell motility and adhesion, we aimed to elucidate the functional consequences of this interaction on cellular processes relevant to breast carcinoma. We demonstrate that this complex slightly inhibits cell attachment and has little effect on protein synthesis in MCF-7 breast cancer cells. However, prebinding IGF-II to immobilized VN was found to significantly enhance breast cancer cell migration through Transwells. Interestingly, IGF-II bound to VN, and not IGF-II in solution in the presence of VN, seems to be responsible for the effects on cell migration. Furthermore, studies using analogs of IGF-II with reduced affinity for the IGF-1R or IGF binding proteins indicate that this response involves the IGF-1R but is independent of IGF binding proteins. This is the first study demonstrating that IGF-II:VN complexes enhance migration of cells. This may prove to be especially relevant, given that overexpression of IGF-II and VN are features of many tumors. PMID- 12746302 TI - The calcium component of gonadotropin-releasing hormone-stimulated luteinizing hormone subunit gene transcription is mediated by calcium/calmodulin-dependent protein kinase type II. AB - Calcium influx plays a critical role in GnRH regulation of rat LH subunit gene transcription, but the site(s) of action are undefined. We investigated the potential of GnRH acting through calcium to activate calcium/calmodulin-dependent protein kinase type II (Ca/CaMK II) in mouse gonadotrope-derived LbetaT2 cells. GnRH stimulated Ca/CaMK II beta subunit activity 3-fold 2 min after treatment and returned to control values by 45 min. The Ca/CaMK II response to GnRH was blocked by administration of the Ca/CaMK II-specific inhibitor, KN-93. The calcium channel activator Bay K 8644 stimulated a 3-fold increase in Ca/CaMK II activity, similar to GnRH. Blocking calcium influx with nimodipine or depleting intracellular calcium storage pools with thapsigargin each resulted in a partial suppression of GnRH-induced activation of Ca/CaMK II, and in combination, completely suppressed the Ca/CaMK II response to GnRH. KN-93 and nimodipine also suppressed alpha-subunit and LHbeta promoter responses to GnRH by 40-60%. LHbeta promoter constructs containing either proximal or proximal and distal GnRH responsive regions were sensitive to inhibition. These data show for the first time that Ca/CaMK II activation plays an important role in the transmission of GnRH signals from the plasma membrane to the LH subunit genes. PMID- 12746305 TI - Lipoprotein enhancement of ovarian theca-interstitial cell steroidogenesis: relative contribution of scavenger receptor class B (type I) and adenosine 5' triphosphate- binding cassette (type A1) transporter in high-density lipoprotein cholesterol transport and androgen synthesis. AB - The theca-interstitial cells take up plasma high-density lipoprotein (HDL)- and low-density-lipoprotein-derived cholesterol to convert into steroid hormones. The uptake of HDL-derived cholesterol is mediated by the scavenger receptor, class B, type I (SR-BI). In nonsteroidogenic cells, HDL-stimulated efflux of cholesterol has been shown to be mediated by the ATP-binding cassette A1 (ABCA1) transporter. Its expression has not been documented in steroidogenic cells. The goal of the present study was to determine: 1) the role of SR-BI in theca-interstitial cell androgen production; 2) whether theca-interstitial cells express ABCA1 transporter mRNA; and 3) the relative roles of SR-BI and ABCA1 transporter in androgen production. The ABCA1 transporter mRNA expression in rat theca interstitial cells was shown using RT-PCR and Northern blot analyses. The role of SR-BI and ABCA1 in androstenedione production was also examined by treating cells with anti-SR-BI and 2-hydroxypropyl-beta-cyclodextrin in the presence and absence of human chorionic gonadotropin and/or human HDL(3). The treatment of theca interstitial cells with anti-SR-BI antibody blocked more than 90% of HDL plus human chorionic gonadotropin-stimulated androstenedione production, and selective HDL-CE uptake. On the other hand, the use of inhibitors of ABCA1 transporter function had no discernible effect on HDL-supported androgen production. These data demonstrate that, although theca-interstitial cells express both SR-BI and ABCA1 transporter mRNA, the SR-BI pathway supplies the majority of the cholesterol required for androgen production. Furthermore, the present study presents evidence for a crucial role for SR-BI in HDL-mediated androgen production. PMID- 12746304 TI - Estrogen-like activity of metals in MCF-7 breast cancer cells. AB - The ability of metals to activate estrogen receptor-alpha (ERalpha) was measured in the human breast cancer cell line, MCF-7. Similar to estradiol, treatment of cells with the divalent metals copper, cobalt, nickel, lead, mercury, tin, and chromium or with the metal anion vanadate stimulated cell proliferation; by d 6, there was a 2- to 5-fold increase in cell number. The metals also decreased the concentration of ERalpha protein and mRNA by 40-60% and induced expression of the estrogen-regulated genes progesterone receptor and pS2 by1.6- to 4-fold. Furthermore, there was a 2- to 4-fold increase in chloramphenicol acetyltransferase activity after treatment with the metals in COS-1 cells transiently cotransfected with the wild-type receptor and an estrogen-responsive chloramphenicol acetyltransferase reporter gene. The ability of the metals to alter gene expression was blocked by an antiestrogen, suggesting that the activity of these compounds is mediated by ERalpha. In binding assays the metals blocked the binding of estradiol to the receptor without altering the apparent binding affinity of the hormone (K(d) = 10(-10) M). Scatchard analysis employing either recombinant ERalpha or extracts from MCF-7 cells demonstrated that (57)Co and (63)Ni bind to ERalpha with equilibrium dissociation constants of 3 and 9.5 x 10(-9) and 2 and 7 x 10(-9) M, respectively. The ability of the metals to activate a chimeric receptor containing the hormone-binding domain of ERalpha suggests that their effects are mediated through the hormone-binding domain. Mutational analysis identified amino acids C381, C447, E523, H524, N532, and D538 as potential interaction sites, suggesting that divalent metals and metal anions activate ERalpha through the formation of a complex within the hormone-binding domain of the receptor. PMID- 12746306 TI - Reversal of behavioral and metabolic abnormalities, and insulin resistance syndrome, by dietary restriction in mice deficient in brain-derived neurotrophic factor. AB - Dietary restriction (DR) extends life span and improves glucose metabolism in mammals. Recent studies have shown that DR stimulates the production of brain derived neurotrophic factor (BDNF) in brain cells, which may mediate neuroprotective and neurogenic actions of DR. Other studies have suggested a role for central BDNF signaling in the regulation of glucose metabolism and body weight. BDNF heterozygous knockout (BDNF+/-) mice are obese and exhibit features of insulin resistance. We now report that an intermittent fasting DR regimen reverses several abnormal phenotypes of BDNF(+/-) mice including obesity, hyperphagia, and increased locomotor activity. DR increases BDNF levels in the brains of BDNF(+/-) mice to the level of wild-type mice fed ad libitum. BDNF(+/-) mice exhibit an insulin-resistance syndrome phenotype characterized by elevated levels of circulating glucose, insulin, and leptin; DR reduces levels of each of these three factors. DR normalizes blood glucose responses in glucose tolerance and insulin tolerance tests in the BDNF(+/-) mice. These findings suggest that BDNF is a major regulator of energy metabolism and that beneficial effects of DR on glucose metabolism are mediated, in part, by BDNF signaling. Dietary and pharmacological manipulations of BDNF signaling may prove useful in the prevention and treatment of obesity and insulin resistance syndrome-related diseases. PMID- 12746307 TI - Neuroimmune response to endogenous and exogenous pyrogens is differently modulated by sex steroids. AB - The objective of this study was to explore whether and how ovarian hormones interact with the febrile response to pyrogens. Estrogen and progesterone treatment of ovariectomized rats was associated with a reduction in lipopolysaccharide (LPS)-induced fever, compared with ovariectomized controls. LPS-fever reduction was accompanied by reduced levels of the inducible cyclooxygenase-2 (COX-2) protein expression in the hypothalamus as well as reduced plasma levels of IL-1beta. The amount of LPS-induced IL-6 in the plasma was not affected by ovarian hormone replacement. In contrast, hypothalamic COX-2 expression in response to intraperitoneal injection of IL-1beta was potentiated by the ovarian hormone replacement. IL-1beta induced a moderate increase in plasma levels of IL-6 that was suppressed by ovarian hormone replacement. These data suggest that ovarian hormone replacement attenuated the proinflammatory response to LPS by suppressing the LPS-induced IL-1beta production and COX-2 expression in the hypothalamus. The markedly different action of ovarian hormones on IL-1beta and LPS effects suggests that this sex hormone modulation of the immune response is a function of the nature of infection and provides further evidence that LPS actions are different from those of IL-1beta. PMID- 12746308 TI - Transcription factors may frame Aa-nat gene expression and melatonin synthesis at night in the Syrian hamster pineal gland. AB - Pineal melatonin synthesis is stimulated at night following an increase in arylalkylamine-N-acetyltransferase (AA-NAT) activity. Depending on the species, two mechanisms of enzyme activation have been described: a cAMP/phospho-cAMP response element-binding protein-dependent stimulation of Aa-nat gene transcription in the rat, presumed to occur in all rodents, or a posttranslational regulation of AA-NAT protein in ongulates. The present data obtained in the Syrian hamster indicate another route of AA-NAT regulation. Elevated nocturnal levels of Aa-nat mRNA were strongly suppressed following light exposure or adrenergic antagonist administration, demonstrating the involvement of norepinephrine in the stimulation of melatonin synthesis. However, administration of adrenergic agonists during the day did not increase Aa-nat mRNA unless a protein synthesis inhibitor was given during the previous night. This indicates that an inhibitory protein, synthesized at night, prevents melatonin synthesis during the day. By contrast, a protein synthesis inhibitor given at the beginning of the night markedly reduced Aa-nat mRNA, suggesting that a stimulatory protein (transcription factor?) is necessary for Aa-nat gene transcription at night. Noteworthy, hamsters raised in long photoperiod were responsive to adrenergic agonist injection only in the first hour after light onset, a response that may be important in this photoperiodic species in which the melatonin peak extends into the morning hours in a short photoperiod. PMID- 12746309 TI - No stress response of the hypothalamo-pituitary-adrenal axis in parturient rats: lack of involvement of brain oxytocin. AB - During parturition, the basal activity of the hypothalamo-pituitary-adrenal (HPA) axis of Wistar rats is strongly attenuated, whereas the oxytocin system is activated. We investigated the secretory responses of the HPA axis and oxytocin to exposure to a mild emotional stressor (airpuff) comparing virgin female, d 22 pregnant, and parturient rats. Furthermore, as the brain oxytocin system is activated in parturition and oxytocin has been shown to inhibit HPA axis responses in virgin rats, the role of brain oxytocin in the regulation of stress responses during parturition was investigated by intracerebroventricular administration of an oxytocin receptor antagonist before stressor exposure (0.75 micro g/5 micro l). In virgin female rats, exposure to airpuff increased ACTH (2.5 +/- 0.34-fold) and corticosterone (5.1 +/- 2.3-fold) secretion, but in late pregnancy and parturition, the stress-induced increase in ACTH (pregnancy: 1.9 +/ 0.41-fold; parturition: 1.3 +/- 0.13-fold) and corticosterone secretion (parturition: 1.8 +/- 0.40-fold) were strongly attenuated. Oxytocin secretion remained unchanged in response to airpuff in both virgin and parturient rats despite higher overall plasma concentrations in the latter. Oxytocin receptor blockade in the brain elevated basal and stress-induced ACTH secretion in virgin but not pregnant or parturient rats and had no effect on oxytocin secretion either in virgin or parturient rats. We conclude that the reactivity of the HPA axis to external stressors is strongly attenuated during parturition, and this cannot be disinhibited by blocking the receptor-mediated action of brain oxytocin. PMID- 12746310 TI - Thyroid hormone enhances aggrecanase-2/ADAM-TS5 expression and proteoglycan degradation in growth plate cartilage. AB - Effects of thyroid hormone on proteoglycan degradation in various regions of cartilage were investigated. In propylthiouracil-treated rats with hypothyroidism, proteoglycan degradation in epiphyseal cartilage during endochondral ossification was markedly suppressed. However, injections of T(4) reversed this effect of propylthiouracil on proteoglycan degradation. In pig growth plate explants, T(3) also induced breakdown of proteoglycan. T(3) increased the release of aggrecan monomer and core protein from the explants into the medium. Accordingly, the level of aggrecan monomer remaining in the tissue decreased after T(3) treatment, and the monomer lost hyaluronic acid-binding capacity, suggesting that the cleavage site is in the interglobular domain. The aggrecan fragment released from the T(3)-exposed explants underwent cleavage at Glu(373)-Ala(374), the major aggrecanase-cleavage site. The stimulation of proteoglycan degradation by T(3) was less prominent in resting cartilage explants than in growth plate explants and was barely detectable in articular cartilage explants. Using rabbit growth plate chondrocyte cultures, we explored proteases that may be involved in T(3)-induced aggrecan degradation and found that T(3) enhanced the expression of aggrecanase-2/ADAM-TS5 (a disintegrin and a metalloproteinase domain with thrombospondin type I domains) mRNA, whereas we could not detect any enhancement of stromelysin, gelatinase, or collagenase activities or any aggrecanase-1/ADAM-TS4 mRNA expression. We also found that the aggrecanse-2 mRNA level, but not aggrecanase-1, increased at the hypertrophic stage during endochondral ossification. These findings suggest that aggrecanse 2/ADAM-TS5 is involved in aggrecan breakdown during endochondral ossification, and that thyroid hormone stimulates the aggrecan breakdown partly via the enhancement of aggrecanase-2/ADAM-TS5. PMID- 12746311 TI - Control of insulin-like growth factor binding protein-5 protease synthesis and secretion by human fibroblasts and porcine aortic smooth muscle cells. AB - IGF binding protein-5 (IGFBP-5) is an important trophic factor for controlling the actions of IGF-I in human dermal fibroblasts and porcine aortic smooth muscle cells. When IGFBP-5 is associated with extracellular matrix, it acts to enhance the cell growth response to IGF-I. The amount of IGFBP-5 within the extracellular matrix is related in part to the amount that is present in conditioned medium, which is related to its rate of synthesis and degradation. A serine protease that degrades IGFBP-5 is present in the conditioned medium of both of these cell types. Because the IGFBP-5 protease activity that is secreted by fibroblasts has been shown to be due to the complement components C1r and C1s, these studies were undertaken to determine whether smooth muscle cells also secreted these proteases and to identify some of the factors that regulate their secretion by both cell types. Both smooth muscle cells and human fibroblasts were shown to release C1r and C1s into conditioned medium. Both C1r and C1s were detected as activated forms, as determined by SDS-PAGE using reducing conditions. The addition of increasing concentrations of either IL-1beta or TNFalpha resulted in increased synthesis of C1s by fibroblasts and smooth muscle cells, and they each increased C1r release. TNFalpha (50 ng/ml) and IL-1beta (20 ng/ml) resulted in maximum stimulation of release of both proteases. In contrast dexamethasone (10(-7) M) had no effect on C1s release and stimulated C1r release only by smooth muscle cells. To determine the physiological significance of this increase in C1r and C1s, the amount of IGFBP-5 protease activity that was present in conditioned medium was determined before and after exposure to TNFalpha, IL-beta, and dexamethasone. All three compounds resulted in an increase in the amount of IGFBP 5 proteolytic activity. Dexamethasone inhibited the release of C(1) inhibitor from fibroblasts, and this contributed to the net increase in proteolytic activity. TNFalpha inhibited the smooth muscle cell DNA synthesis response to IGF I, but the effect of IGF-I was partially restored by the addition of C1 inhibitor. In conclusion, both C1r and C1s are released by cultured fibroblasts, and the release of each into fibroblast or porcine smooth muscle cells medium is stimulated by TNFalpha and IL-1beta. This increase results in a net increase in IGFBP-5 proteolysis, which has the potential to modify IGF-I and IGFBP-5 actions. PMID- 12746312 TI - Vitamin D and dexamethasone inversely regulate parathyroid hormone-induced regulator of G protein signaling-2 expression in osteoblast-like cells. AB - The PTH/PTHrP receptor stimulates both adenylate cyclase- and phospholipase C dependent signaling pathways via different G proteins. The biological actions of PTH on bone are modified by steroid hormones. PTH induces expression of regulator of G protein signaling (RGS)-2, a putative preferential inhibitor of G(q) mediated phospholipase C activation. We investigated whether steroid hormones interfere with PTH signaling by modulating PTH-induced RGS-2 expression in osteoblast-like UMR 106-01 cells. PTH (1-34) rapidly and transiently induced expression of RGS-2 mRNA and protein via the cAMP/protein kinase A pathway within 30 min, with maximal protein abundance after 2 h. PTH-induced RGS-2 preferentially bound to Galpha(q), compared with Galpha(s) protein. 1,25 (OH)(2)D(3) pretreatment enhanced PTH-induced RGS-2 mRNA and protein accumulation, whereas dexamethasone preincubation had an attenuating effect. These effects were due to modulation of the RGS-2 gene transcription rate, which increased by 35% with 1,25-(OH)(2)D(3) and decreased by 63% with dexamethasone pretreatment. RGS-2 mRNA half-life was not affected by either steroid. The transcriptional effects of dexamethasone and 1,25-(OH)(2)D(3) were independent of PTH/PTHrP receptor activation and were not explained by effects on cAMP accumulation, cAMP response element-binding protein expression or phosphorylation, or the abundance of the osteoblast-specific transcription factor core-binding factor alpha (CBFa1/Runx2), a known activator of RGS-2 expression. In conclusion, glucocorticoids and 1,25-(OH)(2)D(3) inversely modulate PTH induced RGS-2 gene transcription. Regulation of RGS-2 may constitute a novel mechanism by which steroids modulate signaling via the PTH/PTHrP receptor and other G protein-coupled receptors in bone. PMID- 12746313 TI - Substitution of cysteine for selenocysteine in the catalytic center of type III iodothyronine deiodinase reduces catalytic efficiency and alters substrate preference. AB - Human type III iodothyronine deiodinase (D3) catalyzes the conversion of T(4) to rT(3) and of T(3) to 3, 3'-diiodothyronine (T2) by inner-ring deiodination. Like types I and II iodothyronine deiodinases, D3 protein contains selenocysteine (SeC) in the highly conserved core catalytic center at amino acid position 144. To evaluate the contribution of SeC144 to the catalytic properties of D3 enzyme, we generated mutants in which cysteine (D3Cys) or alanine (D3Ala) replaces SeC144 (D3wt). COS cells were transfected with expression vectors encoding D3wt, D3Cys, or D3Ala protein. Kinetic analysis was performed on homogenates with dithiothreitol as reducing cofactor. The Michaelis constant of T(3) was 5-fold higher for D3Cys than for D3wt protein. In contrast, the Michaelis constant of T(4) increased 100-fold. The D3Ala protein was enzymatically inactive. Semiquantitative immunoblotting of homogenates with a D3 antiserum revealed that about 50-fold higher amounts of D3Cys and D3Ala protein are expressed relative to D3wt protein. The relative substrate turnover number of D3Cys is 2-fold reduced for T(3) and 6-fold reduced for T(4) deiodination, compared with D3wt enzyme. Studies in intact COS cells expressing D3wt or D3Cys showed that the D3Cys enzyme is also active under in situ conditions. In conclusion, the SeC residue in the catalytic center of D3 is essential for efficient inner-ring deiodination of T(3) and in particular T(4) at physiological substrate concentrations. PMID- 12746315 TI - Expression and characterization of the extracellular Ca(2+)-sensing receptor in melanotrope cells of Xenopus laevis. AB - The extracellular Ca(2+)-sensing receptor (CaR) is expressed in many different organs in various species, ranging from mammals to fish. In some of these organs, this G protein-coupled receptor is involved in the control of systemic Ca(2+) homeostasis, whereas in other organs its role is unclear (e.g. in the pituitary gland). We have characterized the CaR in the neuroendocrine melanotrope cell of the intermediate pituitary lobe of the South African clawed toad Xenopus laevis. First, the presence of CaR mRNA was demonstrated by RT-PCR and in situ hybridization. Then it was shown that activation of the CaR by an elevated extracellular Ca(2+) concentration and different CaR-activators, including L phenylalanine and spermine, stimulates both Ca(2+) oscillations and secretion from the melanotrope. Furthermore, it was revealed that activation of the receptor stimulates Ca(2+) oscillations through opening of voltage-operated Ca(2+) channels in the plasma membrane of the melanotropes. Finally, it was shown that the CaR activator L-phenylalanine could induce the biosynthesis of proopiomelanocortin in the intermediate lobe. Thus, in this study it is demonstrated that the CaR is present and functional in a defined cell type of the pituitary gland, the amphibian melanotrope cell. PMID- 12746314 TI - Retinoic acid stimulates chondrocyte differentiation and enhances bone morphogenetic protein effects through induction of Smad1 and Smad5. AB - Whereas bone morphogenetic protein (BMP)-signaling events induce maturational characteristics in vitro, recent evidence suggests that the effects of other regulators might be mediated through BMP-signaling events. The present study examines the mechanism through which retinoic acid (RA) stimulates differentiation in chicken embryonic caudal sternal chondrocyte cultures. Both RA and BMP-2 induced expression of the chondrocyte maturational marker, colX, in chondrocyte cultures by 8 d. Though the RA effect was small, it synergistically enhanced the effect of BMP-2 on colX and phosphatase activity. Inhibition of either RA or BMP signaling, with selective inhibitors, interfered with the inductive effects of these agents but also inhibited the complementary pathway, demonstrating a codependence of RA and BMP signaling during chondrocyte maturation. BMP-2 did not enhance the effects of RA on an RA-responsive reporter construct, but RA enhanced basal activity and synergistically enhanced BMP-2 stimulation of the BMP-responsive chicken type X collagen reporter. A similar synergistic interaction between RA and BMP-2 was observed on colX expression. RA did not increase the expression of the type IA BMP receptor but did markedly up regulate the expression of Smad1 and Smad5 proteins, important participants in the BMP pathway. Inhibition of RA signaling, with the selective inhibitor AGN 193109, blocked RA-mediated induction of the Smad proteins and chondrocyte differentiation. These findings demonstrate that RA induces the expression of BMP signaling molecules and enhances BMP effects in chondrocytes. PMID- 12746316 TI - 11beta-hydroxysteroid dehydrogenase complementary deoxyribonucleic acid in rainbow trout: cloning, sites of expression, and seasonal changes in gonads. AB - 11beta-Hydroxysteroid dehydrogenases (11beta-HSDs) are important steroidogenic enzymes for catalyzing the interconversion of active glucocorticoid (cortisol and corticosterone) and inert 11-keto forms (cortisone and 11-dehydrocorticosterone) in mammals. In teleosts, 11beta-HSD also plays a role in the production of the predominant androgen, 11-ketotestosterone, in male fish. In this study we cloned cDNAs encoding rainbow trout 11beta-HSD (rt11beta-HSD) from testes and head kidney. The predicted amino acid sequence, hydrophobicity analysis, and transient transfection assays with rt11beta-HSD in HEK293 cells showed that rt11beta-HSD is a homolog of mammalian 11beta-HSD type 2. rt11beta-HSD transcripts are present in steroidogenic tissues and in a number of other tissues. Strong in situ hybridization signals for rt11beta-HSD transcripts were found in Leydig cells of testes, in thecal cells of the early vitellogenic ovarian follicles, and in thecal and granulosa cells of the midvitellogenic and postovulatory follicles. Weaker signals were also found in head kidney interrenal cells from juvenile rainbow trout. Seasonal changes in rt11beta-HSD transcripts in testes showed a pattern similar to that of stress-induced serum cortisol levels, but not to serum androgen levels. High levels of rt11beta-HSD transcripts were found in ovarian follicles from late vitellogenesis through ovulation. These results raise the possibility of a role for rt11beta-HSD in the protection of developing gonads from the inhibitory effects of stress-induced cortisol. PMID- 12746317 TI - C-terminal Src kinase (CSK) modulates insulin-like growth factor-I signaling through Src in 3T3-L1 differentiation. AB - IGF-I stimulates both proliferation and differentiation of adipocyte-precursor cells, preadipocytes in vivo and in vitro. We have previously shown that IGF-I stimulates proliferation of 3T3-L1 preadipocytes through activation of MAPK and MAPK activation by IGF-I is mediated through the Src family of nonreceptor tyrosine kinases. In addition, we have shown that when 3T3-L1 cells reach growth arrest and are stimulated to differentiate, IGF-I can no longer activate the MAPK pathway. We hypothesized that the loss of IGF-I signaling to MAPK in differentiating 3T3-L1 cells is due to loss of IGF-I activation of Src family kinases. We measured c-Src kinase activity in cell lysates from proliferating, growth-arrested and differentiating 3T3-L1 cells. Src activity increased 2- to 4 fold in IGF-I-stimulated proliferating cells; however, IGF-I had a marginal affect on Src activity in growth-arrested cells and inhibited Src activity localized at the membrane in differentiating cells. C-terminal Src kinase (CSK), a ubiquitously expressed nonreceptor tyrosine kinase, negatively regulates the Src family kinases by phosphorylation of the Src C-terminal tyrosine. IGF-I decreased phosphorylation of the Src C-terminal tyrosine in proliferating cells and increased phosphorylation of this site in differentiating cells. IGF-I stimulated CSK kinase activity 2-fold in differentiating 3T3-L1 cells. An association between CSK and c-Src was detected by immunoprecipitation following IGF-I stimulation of differentiating but not proliferating 3T3-L1 cells. These results suggest that the loss of IGF-I downstream mitogenic signaling in differentiating 3T3-L1 cells is due to a change in IGF-I activation of c-Src and CSK may mediate the inactivation of c-Src by IGF-I in 3T3-L1 adipogenesis. PMID- 12746319 TI - Mechanism of adult primitive mesenchymal ST-13 preadipocyte differentiation. AB - Convincing evidence supports the idea that adipogenesis occurs throughout the life of organisms. However, little is known about the adipogenesis program for adult adipocytes. We examine this issue using mouse adult primitive mesenchymal ST-13 preadipocytes that express the peroxisome proliferator-activated receptor gamma (PPARgamma) gene while in a predifferentiated state. The gene expression of PPARgamma was sustained throughout differentiation when ST-13 preadipocytes were induced to become adipocytes by a PPARgamma ligand. However, the differentiation of pluripotent C3H10T1/2 stem cells and 3T3-L1 embryonic fibroblastic cells was associated with enhanced expression of the PPARgamma gene. Immunoblotting analysis revealed that C3H10T1/2 and 3T3-L1 cells expressed low levels of PPARgamma1 from the early stage, and the amount increased during differentiation, whereas PPARgamma2 appeared at the late stage. In contrast, ST-13 preadipocytes expressed an appreciable amount of PPARgamma1 that significantly decreased on differentiation, and a small amount of PPARgamma2 appeared late in the differentiation process. Furthermore, the standard hormone cocktail containing dexamethasone, methylisobutylxanthine, and insulin induced an increase in PPARgamma1 protein only at the early stage, and a low level of PPARgamma2 protein appeared late in ST-13 cells. However, levels of both PPARgamma1 and PPARgamma2 proteins were significantly induced within 2 d in 3T3-L1 cells in this hormonal adipogenesis. Moreover, exposing ST-13 preadipocytes to dexamethasone and insulin induced differentiation, but failed to induce adipogenesis in 3T3-L1. Adipogenesis in adult rat primary preadipocytes was also induced in a similar manner to that of ST-13. Our results indicate that ST-13 cells and primary preadipocytes derived from adults possess an adipogenesis program distinct from that of 3T3-L1 and C3H10T1/2 cells, and that it may represent the adipogenesis program for adult-specific adipocytes. PMID- 12746318 TI - Dexamethasone differentially inhibits thyroxine- or growth hormone-induced body and organ growth of Snell dwarf mice. AB - Supraphysiological doses of glucocorticoids cause growth retardation in both animals and humans. Many studies have addressed the interaction of glucocorticoids with the GH/IGF system, but little is known about the effect of glucocorticoids on T(4)-stimulated growth. The Snell dwarf mouse is deficient in GH, thyroid-stimulating hormone, and prolactin and therefore allows the study of the effect of glucocorticoids on the growth induced by GH and T(4) without their mutual interaction. Four weeks of treatment with T(4) (1 micro g/d) or human GH (50 mU/d) equally increased nose-tail length (3.1 +/- 0.1 cm and 3.0 +/- 0.2 cm, respectively). Dexamethasone (DXM) had much less impact on T(4)-stimulated growth than on GH-induced growth (T(4) + DXM: 2.4 +/- 0.1 cm vs. GH+ DXM: 1.4 +/- 0.1 cm). Similar data were obtained for body weight gain. T4 and GH had a different effect on the weight of various organs: GH caused a higher increase in liver and lumbar vertebrae weight, and T(4) was a better stimulator for kidney (P < 0.05), thymus, and spleen growth. Remarkably, T(4)-stimulated growth of the organs was less affected by DXM than GH-induced organ growth. GH even potentiated the growth inhibition by DXM in the thymus and tibia. In conclusion, T(4)-stimulated growth in Snell dwarf mice is less affected by DXM than growth stimulated by GH PMID- 12746320 TI - Overexpression of glutamic acid decarboxylase-67 (GAD-67) in gonadotropin releasing hormone neurons disrupts migratory fate and female reproductive function in mice. AB - gamma-Aminobutyric acid (GABA) inhibits the embryonic migration of GnRH neurons and regulates hypothalamic GnRH release. A subset of GnRH neurons expresses GABA along their migratory route in the nasal compartment before entering the brain, suggesting that GABA produced by GnRH neurons may help regulate the migratory process. To examine this hypothesis and the possibility that persistence of GABA production by GnRH neurons may affect subsequent reproductive function, we generated transgenic mice in which the expression of glutamic acid decarboxylase 67 (GAD-67), a key enzyme in GABA synthesis, is targeted to GnRH neurons under the control of the GnRH gene promoter. On embryonic d 15, when GnRH neurons are still migrating, the transgenic animals had more GnRH neurons in aberrant locations in the cerebral cortex and fewer neurons reaching the hypothalamic preoptic region, whereas migration into the brain was not affected. Hypothalamic GnRH content in mutant mice was low during the first week of postnatal life, increasing to normal values during infantile development (second week after birth) in the presence of increased pulsatile GnRH release. Consistent with these changes, serum LH and FSH levels were also elevated. Gonadotropin release returned to normal values by the time steroid negative feedback became established (fourth week of life). Ovariectomy at this time demonstrated an enhanced gonadotropin response in transgenic animals. Although the onset of puberty, as assessed by the age at vaginal opening and first ovulation, was not affected in the mutant mice, estrous cyclicity and adult reproductive capacity were disrupted. Mutant mice had reduced litter sizes, increased time intervals between deliveries of litters, and a shorter reproductive life span. Thus, GABA produced within GnRH neurons does not delay GnRH neuronal migration, but instead serves as a developmental cue that increases the positional diversity of these neurons within the basal forebrain. In addition, the results suggest that the timely termination of GABA production within the GnRH neuronal network is a prerequisite for normal reproductive function. The possibility arises that similar abnormalities in GABA homeostasis may contribute to syndromes of hypothalamic amenorrhea/oligomenorrhea in humans. PMID- 12746322 TI - Osteonectin-null mutation compromises osteoblast formation, maturation, and survival. AB - Osteonectin, also known as SPARC (secreted protein acidic and rich in cysteine) or BM-40, is one of the most abundant noncollagenous proteins in bone. Analysis of osteonectin-null mice revealed that osteonectin is necessary for the maintenance of bone mass and normal remodeling, as osteonectin-null mice have decreased osteoblast number and bone formation rate. Cultures of bone marrow stromal cells and osteoblasts from control and osteonectin-null mice were used to determine the cellular basis for the mutant phenotype. We found that marrow stroma from osteonectin-null mice contains fewer osteoblastic precursors than that of control mice, and the osteonectin-null mutation did not affect the proliferation rate of stromal cells or osteoblasts. Whereas osteonectin-null cells could adopt an osteoblastic phenotype, a smaller proportion of these cells expressed markers of a fully differentiated osteoblast. Mutant cells exhibited decreased formation of mineralized nodules, as well as diminished expression of osteocalcin mRNA and response to PTH. Furthermore, osteonectin-null cells showed an increased tendency to form adipocytes, with enhanced expression of the adipocytic markers adipsin and CCAAT/enhancer binding protein delta. Osteonectin null cells were also more susceptible to environmental stresses. These data indicate that osteonectin is important for osteoblast formation, maturation, and survival. PMID- 12746321 TI - Corticotropin-releasing factor receptor-2-deficient mice display abnormal homeostatic responses to challenges of increased dietary fat and cold. AB - Corticotropin-releasing factor (CRF) and its family of ligands are key regulators of energy balance. These ligands function via activation of their two receptors, CRFR1 and CRFR2. CRFR1 has been shown to be the dominant receptor in activation of the hypothalamic-pituitary-adrenal axis in response to stress as well as a key mediator of anxiety in the limbic system. To specifically examine the role of CRFR2 in energy balance, mice deficient for CRFR2 were exposed to physiological perturbations of homeostasis, including high-fat diet, repeated cold stress, and glucose and insulin challenges, and their responses measured. While on a high-fat diet, CRFR2-mutant mice consumed substantially more food and maintaining the same weight but had significantly lower body fat and lower plasma lipids than their wild-type littermates. These mice were also less inclined to develop diet-induced insulin resistance and more sensitive to changes in plasma glucose, indicating increased insulin sensitivity. Following repeated cold stress, mutant mice had significantly lower body fat and a transient reduction in feed efficiency, despite similar body weights, suggesting a possible preference for fat as an energy substrate. Elevated levels of uncoupling protein-1 in brown adipose tissue as well as smaller white and brown adipocytes from CRFR2-mutant mice were indications of possible increased sympathetic tone. These results demonstrate that CRFR2 plays a critical role in regulation of energy expenditure and is important for responses to homeostatic challenges. PMID- 12746323 TI - Expression of a novel C-type lectin in the mouse vagina. AB - Estrogens regulate the proliferation and differentiation of mouse vaginal epithelial cells. However, the molecular mechanisms underlying estrogen-induced changes have not been elucidated. The goal of this study was to identify estrogen responsive genes related to the proliferation and differentiation of mouse vaginal epithelial cells. We used differential display to reveal specific genes regulated by estrogens and identified a transcript that was designated DDV10. DDV10 encodes a membrane protein with a C-type lectin domain in the carboxyl terminal region; thus, we inferred that it belongs to the C-type lectin family. We analyzed the temporal and spatial expression of DDV10 using RT-PCR, quantitative real-time RT-PCR, and in situ hybridization. Ovariectomy decreased DDV10 mRNA levels, whereas 17beta-estradiol treatment increased expression of DDV10 mRNA in vaginas of ovariectomized mice. DDV10 mRNA was first detected between 20 and 30 d after birth and was found in eye, tongue, stomach, and stratified and cornified vaginal epithelial cells, but not in stromal cells or uterus. DDV10 transcripts were not detected in vaginas of estrogen receptor alpha knockout mice. Taken together, these data suggest that DDV10 encodes a novel, 17beta-estradiol-regulated, C-type lectin in the mouse vagina. DDV10 may play a role in the stratification and/or cornification of epithelial cells during differentiation. PMID- 12746324 TI - Mouse endocrine gland-derived vascular endothelial growth factor: a distinct expression pattern from its human ortholog suggests different roles as a regulator of organ-specific angiogenesis. AB - We recently described human endocrine gland-derived vascular endothelial growth factor (EG-VEGF) as an endothelial cell mitogen with a novel selective activity and an expression pattern essentially limited to steroidogenic glands. Herein we present the identification and characterization of the mouse ortholog. The mouse cDNA and predicted amino acid sequences are, respectively, 86% and 88% identical with the human. Surprisingly, the mouse EG-VEGF transcript is predominantly expressed in liver and kidney. A comparison of human and mouse EG-VEGF promoter sequences revealed a potential binding site for NR5A1, which is known to be a pivotal element for steroidogenic-specific transcription, in the human but not mouse promoter. In situ hybridization studies localized expression of mouse EG VEGF mRNA to hepatocytes and renal tubule cells. Interestingly, capillary endothelial cells in these sites share several common structural features with those found in steroidogenic glands. Within liver and kidney, EG-VEGF receptor expression was largely restricted to endothelial cells. Mouse EG-VEGF promoted proliferation and survival of endothelial cells. We propose that mouse EG-VEGF, like human EG-VEGF, plays a role in regulating the phenotype and growth properties of endothelial cells within distinct capillary beds. PMID- 12746325 TI - Pituitary hormones inhibit the function and differentiation of fetal Sertoli cells. AB - Although the role of pituitary hormones in fetal Sertoli cell proliferation is well understood, their involvement in fetal Sertoli cell differentiation is poorly documented. In this study, we evaluated rat fetal Sertoli cell function by measuring basal transferrin secretion ex vivo and transferrin and anti-Mullerian hormone (AMH) mRNA levels in vivo. The differentiation state of the Sertoli cells was estimated from the amount of transferrin secreted ex vivo after acute stimulation with FSH. Surprisingly, we found that the amount of transferrin secreted by each Sertoli cell in basal condition and after acute FSH stimulation decreased between 18.5 and 21.5 day post coitum (dpc), which corresponds to the onset of pituitary hormone secretion. All of the Sertoli cell parameters measured (basal and FSH-stimulated transferrin secretion ex vivo, transferrin and AMH mRNA levels in vivo) were higher in 21.5-dpc fetuses that had been decapitated on 16.5 dpc than in control littermates. Furthermore, immunostaining for AMH was strongly increased after decapitation. Taken together, these results suggest that pituitary hormones in the fetus and in the immature or adult rat differently regulate Sertoli cells, which suggests that fetal Sertoli cells have their own particular physiology. PMID- 12746326 TI - Ghrelin inhibits the development of mouse preimplantation embryos in vitro. AB - Although ghrelin acts as a modulator of feeding behavior and energy metabolism in the central nervous system, recent studies have implicated the peripheral actions of ghrelin in reproductive tissues. Here, we investigated the expression of ghrelin and its receptor (GHS-R) in mouse oocyte and preimplantation embryos, and we examined the role of ghrelin in the regulation of early embryo development. Both ghrelin and GHS-R mRNAs were detected in morula or more advanced embryo stages. As for the origin of ghrelin, both ghrelin mRNA and protein were identified in the uterine endometrium. The levels of ghrelin in uterine fluid as well as plasma were significantly increased in fasting mice compared with animals with free access to foods. Addition of ghrelin to culture media inhibited the development of two-cell embryos to the hatched blastocysts, and the inhibitory effects of ghrelin were abolished by an antagonist for the GHS-R. In addition, ghrelin significantly decreased the number of total cells, inner cell mass, and trophectoderm cells in blastocysts. These observations suggest that ghrelin could inhibit the development of preimplantation embryos during fasting. Thus, ghrelin may act as a peripheral factor to avoid the excess metabolic demands imposed by pregnancy during malnutritional states. PMID- 12746327 TI - Changes in hypothalamic expression levels of galanin-like peptide in rat and mouse models support that it is a leptin-target peptide. AB - Galanin-like peptide (GALP) is a novel peptide that has been isolated from the porcine hypothalamus. The expression of GALP mRNA is localized to the hypothalamic arcuate nucleus and is thought to be under the regulation of leptin. First, we confirmed by real-time PCR analysis that sc administration of leptin to Wistar rats under food-deprived conditions resulted in a 1.5-fold increase in hypothalamic GALP mRNA levels. Next, GALP mRNA levels were found to be reduced by 50% in 11-wk-old male Zucker obese rats compared with age-matched Zucker lean rats, whereas neuropeptide Y mRNA levels were increased by 55% and proopiomelanocortin mRNA levels were reduced by 53% in Zucker obese rats. Analysis using a two-site enzyme immunoassay revealed a lower level of hypothalamic GALP immunoreactivity in 11-wk-old Zucker obese rats (5.9 fmol/mg protein) than in age-matched Zucker lean rats (19.6 fmol/mg protein). Immunohistochemical studies demonstrated that Zucker obese rats (11 wk old) had a reduced number of GALP immunoreactivity-positive cells (29.4 cells/3 slices) in the arcuate nucleus compared with age-matched Zucker lean rats (115 cells/3 slices). Furthermore, Zucker obese rats showed increased sensitivity to intracerebroventricularly administered GALP compared with Zucker lean rats, in that a lower dose of GALP increased plasma LH levels in male Zucker obese rats, but not in male Zucker lean rats. In addition, a reduction in the level of hypothalamic GALP mRNA was found in db/db and ob/ob mice. The result supports the hypothesis that the hypothalamic GALP gene expression is controlled by leptin signals and suggests possible involvement of GALP in the reproductive abnormalities of the Zucker obese rat. PMID- 12746328 TI - Committing embryonic stem cells to differentiate into thyrocyte-like cells in vitro. AB - The derivation of thyrocyte-like cells in culture is of importance in the basic study of early thyroid embryogenesis and the generation of an unlimited clinical source of thyrocytes for genetic manipulation and cell transplantation. We have established an experimental system, which shows that 6-d-old embryoid bodies (EBs) differentiated from mouse embryonic stem (ES) cells expressed a set of genes traditionally associated with thyroid cells. The genes analyzed included the thyroid transcription factor PAX8, the Na(+)/I(-) symporter, thyroperoxidase, thyroglobulin, and the TSH receptor (TSHR). Immunofluorescent analysis demonstrated the presence of TSHR-positive cells as outgrowths from 8-d-old EBs cultured on chamber slides. Accordingly, this area of cells also expressed PAX8 and another thyroid transcription factor TTF2. Of importance, TSH, the main regulator of the thyroid gland, was necessary to maintain the expression of PAX8 and TSHR genes during EB differentiation. Furthermore, thyroid-specific function, such as cAMP generation by TSH, was maintained in this model. Together, these results suggested that the developmental program associated with thyrocyte development is recapitulated in the ES/EB model system. The differentiation of mouse ES cells into thyrocyte-like cells provides a powerful model for the study of thyrocyte developmental diseases associated with this lineage and contributes to the development of thyroid hormone-secreting cell lines. PMID- 12746330 TI - Human uridine diphosphate-glucuronosyltransferase UGT2B7 conjugates mineralocorticoid and glucocorticoid metabolites. AB - Mineralocorticoid and glucocorticoid hormones are metabolized as glucuronide conjugates. Using labeled [(14)C]uridine diphosphate glucuronic acid and microsomal preparations from human embryonic kidney 293 cells stably expressing the different human and monkey uridine diphosphate glucuronosyltransferase (UGT)2B enzymes, it is demonstrated that the two human allelic variants UGT2B7H((268)) and UGT2B7Y((268)) conjugate aldosterone, its A-ring reduced metabolites (5alpha-dihydroaldosterone and 3alpha,5beta-tetrahydroaldosterone), and both 5alpha- and 5beta-tetrahydrocortisone epimers. The two variants of UGT2B4 also glucuronidate tetrahydroaldosterone, whereas all enzymes tested were inefficient to produce cortisol glucuronide derivatives. Kinetic analyses reveal that UGT2B7 polymorphisms glucuronidate mineralocorticoids with a 5.5- to 20-fold higher affinity than glucocorticoids. For the first time, a significant difference between the two allelic variants of UGT2B7 is described, because UGT2B7H((268)) possesses an 11-fold higher aldosterone glucuronidation efficiency (ratio Vmax((app.))/Km((app.))) than UGT2B7Y((268)). RT-PCR experiments demonstrate the expression of UGT2B7 in human kidney and in renal proximal tubule epithelial cells, suggesting that mineralocorticoids and glucocorticoids are metabolized in their target tissue. Measurement of aldosterone glucuronidation and normalization with the UGT2B protein contents in monkey tissues demonstrate that liver and kidney glucuronidate this hormone with a similar velocity. Immunohistochemical studies performed in monkey kidney cortex reveal a restrictive expression of UGT2B proteins in the epithelial cells of the proximal tubules. Because expression of the mineralocorticoid receptor was detected in the distal tubule epithelial cells, the present data suggest a two-cell mechanism of aldosterone action and metabolism in the kidney. PMID- 12746329 TI - Signaling differences from the A and B isoforms of the insulin receptor (IR) in 32D cells in the presence or absence of IR substrate-1. AB - The A isoform of the insulin receptor (IR) is frequently overexpressed in cancer cells and is activated by IGF-II as well as by insulin, whereas the B isoform is predominant in differentiated tissues and responds poorly to IGF-II. The IR substrate-1 (IRS-1), a docking protein for the IR, is known to send a mitogenic signal and to be a powerful inhibitor of cell differentiation. We have investigated the biological effects of the two IR isoforms in parental 32D hemopoietic cells, which do not express IRS-1, and in 32D-derived cells in which IRS-1 is ectopically expressed. The effects of the two isoforms on cell survival, differentiation markers and nuclear translocation of IRS-1 were compared. The results confirm that the A isoform responds to IGF-II and preferentially sends mitogenic, antiapoptotic signals, whereas the B form, poorly responsive to IGF II, tends to send differentiation signals. PMID- 12746331 TI - Targeted expression of the dominant-negative prolactin receptor in the mammary gland of transgenic mice results in impaired lactation. AB - The F3-short form of the rat PRL receptor (F3-SPRLR) form acts as a dominant negative inhibitor in vitro. We have developed a transgenic mouse model in which the rat F3-SPRLR was expressed in mammary epithelium under the control of the mouse mammary tumor virus promoter. Two lines of mice were characterized and shown to express the transgene in the mammary gland. No developmental abnormalities or differences from wild-type littermates were observed on the basis of size, activity, or fertility. Mice with a low level of transgene expression had a mammary phenotype similar to the wild type. However, mice overexpressing the transgene (levels much higher than those of the endogenous long PRLR transcript) had impaired mammary gland differentiation and lactation. In these mice, whole-mount and histological analyses demonstrated normal ductal development, but severely reduced lobuloalveolar outgrowth. signal transducer and activator of transcription-5 phosphorylation and expression of beta-casein and whey acidic protein gene were decreased. In vivo bromodeoxyuridine incorporation at midpregnancy showed that the reduction in mammary development was not due to an inhibition of ductal growth and side-branching. This model demonstrates for the first time in vivo a function of the SPRLR and a local and targeted effect of PRL on the mammary gland that are essential for its function, but not for its development. PMID- 12746332 TI - A ketogenic diet increases brain insulin-like growth factor receptor and glucose transporter gene expression. AB - A ketogenic diet suppresses seizure activity in children and in juvenile rats. To investigate whether alteration in brain IGF activity could be involved in the beneficial effects of the ketogenic diet, we examined the effects of this diet on IGF system gene expression in the rat brain. Juvenile rats were fed one of three different diets for 7 d: ad libitum standard rat chow (AL-Std), calorie restricted standard chow (CR-Std), or a calorie-restricted ketogenic diet (CR Ket). The calorie-restricted diets contained 90% of the rats' calculated energy requirements. The AL-Std diet group increased in weight, whereas the two CR groups merely maintained their weight during the 7-d diet. Glucose levels were significantly reduced in both CR groups compared with the AL-Std group, but only the CR-Ket group developed ketonemia. IGF1 mRNA levels were reduced by 30-50% in most brain regions in both CR groups. IGF1 receptor (IGF1R) mRNA levels were decreased in the CR-Std group but were increased in the CR-Ket diet group. Brain IGF binding protein (IGFBP)-2 and -5 mRNA levels were not altered by diet, but IGFBP-3 mRNA levels were markedly increased by the ketogenic diet while not altered by calorie restriction alone. Brain glucose transporter expression was also investigated in this study. Glucose transporter (GLUT) 4 mRNA levels were quite low and not appreciably altered by the different diets. Parenchymal GLUT1 mRNA levels were increased by the CR-Ket diet, but endothelial GLUT1 mRNA levels were not affected. Neuronal GLUT3 expression was decreased with the CR-Std diet and increased with the CR-Ket diet, in parallel with the IGF1R pattern. These observations reveal divergent effects of dietary caloric content and macronutrient composition on brain IGF system and GLUT expression. In addition, the data may be consistent with a role for enhanced IGF1R and GLUT expression in ketogenic diet-induced seizure suppression. PMID- 12746333 TI - Developmental and hormonal signals dramatically alter the localization and abundance of insulin receptor substrate proteins in the mammary gland. AB - Insulin receptor substrates (IRS) are central integrators of hormone, cytokine, and growth factor signaling. IRS proteins can be phosphorylated by a number of signaling pathways critical to normal mammary gland development. Studies in transgenic mice that overexpress IGF-I in the mammary gland suggested that IRS expression is important in the regulation of normal postlactational mammary involution. The goal of these studies was to examine IRS expression in the mouse mammary gland and determine the importance of IRS-1 to mammary development in the virgin mouse. IRS-1 and -2 show distinct patterns of protein expression in the virgin mouse mammary gland, and protein abundance is dramatically increased during pregnancy and lactation, but rapidly lost during involution. Consistent with hormone regulation, IRS-1 protein levels are reduced by ovariectomy, induced by combined treatment with estrogen and progesterone, and vary considerably throughout the estrous cycle. These changes occur without similar changes in mRNA levels, suggesting posttranscriptional control. Mammary glands from IRS-1 null mice have smaller fat pads than wild-type controls, but this reduction is proportional to the overall reduction in body size. Development of the mammary duct (terminal endbuds and branch points) is not altered by the loss of IRS-1, and pregnancy-induced proliferation is not changed. These data indicate that IRS undergo complex developmental and hormonal regulation in the mammary gland, and that IRS-1 is more likely to regulate mammary function in lactating mice than in virgin or pregnant mice. PMID- 12746334 TI - Shortened life span, bradycardia, and hypotension in mice with targeted expression of an Igf2 transgene in smooth muscle cells. AB - IGF2 is known to affect the normal development and pathology of the cardiovascular system. We previously created mutant mice with targeted expression of an Igf2 transgene in the smooth muscle cells and showed that these mice spontaneously develop aortic intimal cushions. In the present work, we provide a general description of the phenotype of two independent lines of heterozygous transgenics. These mice showed organomegaly and a shortened life span. The latter trait was stronger in the line with a relatively more marked organomegaly and more pronounced in males than females in both lines. Postmortem histology revealed gross abnormalities of the cardiac architecture, suggesting that transgenic mice may accumulate lethal cardiovascular defects. Accordingly, apparently normal transgenic mice had mild cardiomegaly, an enlarged left ventricle, bradycardia, and hypotension. These observations are discussed in the light of the proposed therapeutic use of IGF2 in human cardiac diseases. PMID- 12746335 TI - Cloning of a functional vitamin D receptor from the lamprey (Petromyzon marinus), an ancient vertebrate lacking a calcified skeleton and teeth. AB - The nuclear vitamin D receptor (VDR) mediates the actions of its 1,25 dihydroxyvitamin D(3) ligand to control gene expression in terrestrial vertebrates. Prominent functions of VDR-regulated genes are to promote intestinal absorption of calcium and phosphate for bone mineralization and to potentiate the hair cycle in mammals. We report the cloning of VDR from Petromyzon marinus, an unexpected finding because lampreys lack mineralized tissues and hair. Lamprey VDR (lampVDR) clones were obtained via RT-PCR from larval protospleen tissue and skin and mouth of juveniles. LampVDR expressed in transfected mammalian COS-7 cells bound 1,25-dihydroxyvitamin D(3) with high affinity, and transactivated a reporter gene linked to a vitamin D-responsive element from the human CYP3A4 gene, which encodes a P450 enzyme involved in xenobiotic detoxification. In tests with other vitamin D responsive elements, such as that from the rat osteocalcin gene, lampVDR showed little or no activity. Phylogenetic comparisons with nuclear receptors from other vertebrates revealed that lampVDR is a basal member of the VDR grouping, also closely related to the pregnane X receptors and constitutive androstane receptors. We propose that, in this evolutionarily ancient vertebrate, VDR may function in part, like pregnane X receptors and constitutive androstane receptors, to induce P450 enzymes for xenobiotic detoxification. PMID- 12746337 TI - Growth hormone-releasing hormone (GHRH) neurons in GHRH-enhanced green fluorescent protein transgenic mice: a ventral hypothalamic network. AB - The hypothalamic GHRH neurons secrete pulses of GHRH to generate episodic GH secretion, but little is known about the mechanisms involved. We have made transgenic mice expressing enhanced green fluorescent protein (eGFP) specifically targeted to the secretory vesicles in GHRH neurons. GHRH cells transported eGFP from cell bodies in the arcuate nucleus to extensively arborized varicose fiber terminals in the median eminence. Patch clamp recordings from visually identified GHRH cells in mature animals showed spontaneous action potentials, often firing in short bursts up to 10 Hz. GHRH neurons received frequent synaptic inputs, as demonstrated by the recording of abundant inward postsynaptic currents, but spikes were followed by large after-hyperpolarizations, which limited their firing rate. Because many GHRH neurons lie close to the ventral hypothalamic surface, this was examined by wide-field binocular epifluorescence stereomicroscopy. This approach revealed an extensive horizontal network of GHRH cells at low power and individual fiber projections at higher power in the intact brain. It also showed the dense terminal projections of the GHRH cell population in the intact median eminence. This model will enable us to characterize the properties of individual GHRH neurons and their structural and functional connections with other neurons and to study directly the role of the GHRH neuronal network in generating episodic secretion of GH. PMID- 12746336 TI - Specific and combined effects of insulin and glucose on functional pancreatic beta-cell mass in vivo in adult rats. AB - We investigated the specific and associated effects of insulin and glucose on beta-cell growth and function in adult rats. By combining simultaneous infusion either of glucose and/or insulin or glucose and diazoxide, three groups of rats were constituted: hyperglycemic-hyperinsulinemic rats (high glucose-high insulin), hyperglycemic-euinsulinemic rats (high glucose), and euglycemic hyperinsulinemic rats (high insulin). All the infusions lasted 48 h. Control rats were infused with 0.9% NaCl (saline controls). In all groups, beta-cell mass was significantly increased, compared with controls (by 70% in high glucose-high insulin rats, 65% in high glucose rats, and 50% in high insulin rats). The stimulation of neogenesis was suggested by the high number of islets budding from pancreatic ducts in high glucose-high insulin and high glucose rats and by the presence of numerous clusters of few beta-cells within the exocrine pancreas in high insulin rats. beta-Cell hypertrophy was observed only in high glucose-high insulin rats. The rate of beta-cell proliferation was similar to that of controls in high glucose-high insulin rats after a 48-h glucose infusion, dropped dramatically in high insulin rats, and dropped to a lesser extent in high glucose rats. In high glucose-high insulin and high glucose rats, beta-cell mass increase was related to a higher beta-cell responsiveness to glucose in vitro as measured by islet perifusion studies, whereas in high insulin rats, no significant enhancement of glucose induced insulin secretion could be noticed. The data show that glucose and insulin may have specific stimulating effects on beta-cell growth and function in vivo in adult rats independently of the influence they exert each other on their respective plasma concentration. PMID- 12746340 TI - As West Nile virus season heats up, blood safety testing lags behind. PMID- 12746339 TI - A piece of my mind. Alice's husband. PMID- 12746338 TI - Neuronal histamine regulates food intake, adiposity, and uncoupling protein expression in agouti yellow (A(y)/a) obese mice. AB - Hypothalamic neuronal histamine and its H(1) receptor (H(1)-R) form a part of the leptin-signaling pathway in the brain and have been shown to regulate body weight and adiposity in diabetic (db/db) and diet-induced obese mice by affecting food intake and uncoupling protein mRNA expression. The proopiomelanocortin (POMC) melanocortin-4 receptor (MC-4R) is also important for leptin signaling. The present study had two aims: first, to clarify the antiobesity action of neuronal histamine in agouti yellow (A(y)/a) obese mice, a model of obesity in which POMC/MC-4R signaling is disrupted by blockade of MC-4R and second, to investigate the functional relationship between neuronal histamine and POMC/MC-4R signaling. Central administration of histamine into the lateral cerebroventricle decreased cumulative food intake and body weight in A(y)/a obese mice. Histamine treatment also decreased mRNA expression of ob gene in epididymal white adipose tissue and up-regulated uncoupling protein 1 mRNA expression in brown adipose tissue. These effects were attenuated in A(y)/a obese mice with histamine H(1)-receptor (H(1) R) knockout. Histamine treatment induced c-Fos-like immunoreactivity in both paraventricular and arcuate nucleus. There was no significant difference in histamine-induced c-Fos-like immunoreactivity in the hypothalamus between A(y)/a obese mice and lean littermates, indicating histamine signaling was not disrupted at the hypothalamic level in A(y)/a obese mice. These results suggest that neuronal histamine have an antiobese action, even in A(y)/a obese mice despite a deficiency in POMC/MC-4R signaling. In addition, it appears that the histamine H(1)-R signaling pathway may be independent or downstream of the POMC/MC-4R signaling. PMID- 12746341 TI - Experts debate widening use of opioid drugs for chronic nonmalignant pain. PMID- 12746346 TI - From the Centers for Disease Control and Prevention. Update: severe acute respiratory syndrome--United States, 2003. PMID- 12746347 TI - From the Centers for Disease Control and Prevention. Update: adverse events following civilian smallpox vaccination--United States, 2003. PMID- 12746348 TI - From the Centers for Disease Control and Prevention. Prevalence of current cigarette smoking among adults and changes in prevalence of current and some day smoking--United States, 1996-2001. PMID- 12746349 TI - Cost-effectiveness of screening for lung cancer. PMID- 12746350 TI - Cost-effectiveness of screening for lung cancer. PMID- 12746351 TI - Cost-effectiveness of screening for lung cancer. PMID- 12746352 TI - Cholinesterase inhibitors for Alzheimer disease. PMID- 12746353 TI - Cholinesterase inhibitors for Alzheimer disease. PMID- 12746354 TI - Cholinesterase inhibitors for Alzheimer disease. PMID- 12746355 TI - Cholinesterase inhibitors for Alzheimer disease. PMID- 12746356 TI - Ethanol in hospital formularies. PMID- 12746359 TI - Hypertension prevalence and blood pressure levels in 6 European countries, Canada, and the United States. AB - CONTEXT: Geographic variations in cardiovascular disease (CVD) and associated risk factors have been recognized worldwide. However, little attention has been directed to potential differences in hypertension between Europe and North America. OBJECTIVE: To determine whether higher blood pressure (BP) levels and hypertension are more prevalent in Europe than in the United States and Canada. DESIGN, SETTING, AND PARTICIPANTS: Sample surveys that were national in scope and conducted in the 1990s were identified in Germany, Finland, Sweden, England, Spain, Italy, Canada, and the United States. Collaborating investigators provided tabular data in a consistent format by age and sex for persons at least 35 years of age. Population registries were the main basis for sampling. Survey sizes ranged from 1800 to 23 100, with response rates of 61% to 87.5%. The data were analyzed to provide age-specific and age-adjusted estimates of BP and hypertension prevalence by country and region (eg, European vs North American). MAIN OUTCOME MEASURES: Blood pressure levels and prevalence of hypertension in Europe, the United States, and Canada. RESULTS: Average BP was 136/83 mm Hg in the European countries and 127/77 mm Hg in Canada and the United States among men and women combined who were 35 to 74 years of age. This difference already existed among younger persons (35-39 years) in whom treatment was uncommon (ie, 124/78 mm Hg and 115/75 mm Hg, respectively), and the slope with age was steeper in the European countries. For all age groups, BP measurements were lowest in the United States and highest in Germany. The age- and sex-adjusted prevalence of hypertension was 28% in the North American countries and 44% in the European countries at the 140/90 mm Hg threshold. The findings for men and women by region were similar. Hypertension prevalence was strongly correlated with stroke mortality (r = 0.78) and more modestly with total CVD (r = 0.44). CONCLUSIONS: Despite extensive research on geographic patterns of CVD, the 60% higher prevalence of hypertension in Europe compared with the United States and Canada has not been generally appreciated. The implication of this finding for national prevention strategies should be vigorously explored. PMID- 12746360 TI - Prevalence and characteristics of chronic pain among chemically dependent patients in methadone maintenance and residential treatment facilities. AB - CONTEXT: Little is known about the prevalence and characteristics of chronic pain among patients with different types of chemical dependency. OBJECTIVES: To estimate the prevalence and to examine the characteristics of chronic severe pain in chemically dependent populations receiving methadone maintenance or inpatient residential treatment. DESIGN, SETTING, AND PARTICIPANTS: Representative samples of 390 patients from 2 methadone maintenance treatment programs (MMTPs) and 531 patients from 13 short-term residential substance abuse treatment (inpatient) programs, all in New York State, were surveyed in late 2000 and early 2001. MAIN OUTCOME MEASURE: Prevalence of chronic severe pain, defined as pain that persisted for more than 6 months and was of moderate to severe intensity or that significantly interfered with daily activities. RESULTS: Chronic severe pain was experienced by 37% of MMTP patients (95% confidence interval [CI], 32%-41%) and 24% of inpatients (95% CI, 20%-28%; P =.03). Pain of any type or duration during the past week was reported by 80% of MMTP patients and 78% of inpatients. Among those with chronic severe pain, 65% of MMTP patients and 48% of inpatients reported high levels of pain-related interference in physical and psychosocial functioning. Among MMTP patients, correlates of chronic pain in a multivariate model were age (odds ratio [OR], 2.08; 95% CI, 1.17-3.70), chronic illness (OR, 1.88; 95% CI, 1.07-3.29), lifetime psychiatric illness (OR, 1.77; 95% CI, 1.06 2.97), psychiatric distress (OR, 1.63; 95% CI, 1.22-2.18), and time in treatment (OR, 2.23; 95% CI, 1.06-4.68). Among inpatients, the correlates of chronic pain were race (blacks vs whites: OR, 0.52; 95% CI, 0.31-0.90; Hispanics vs whites: OR, 0.48; 95% CI, 0.24-0.95), drug craving (OR, 2.78; 95% CI, 1.54-5.02), chronic illness (OR, 2.17; 95% CI, 1.37-3.43), and psychiatric distress (OR, 1.36; 95% CI, 1.03-1.81). Among those with chronic severe pain, inpatients were significantly more likely than MMTP patients to have used illicit drugs, as well as alcohol, to treat their pain complaint (51% vs 34%, P =.005) but were less likely to have been prescribed pain medications (52% vs 67%, P =.01). CONCLUSIONS: Chronic severe pain is prevalent among patients in substance abuse treatment, especially MMTP patients. Pain is associated with functional impairment and correlates of pain vary with the population. Self-medication for pain with psychoactive drugs appears especially problematic among substance users who enroll in drug-free treatment programs. Substance abuse treatment programs need to develop comprehensive and structured pain management programs. PMID- 12746361 TI - Relationship of changes in physical activity and mortality among older women. AB - CONTEXT: Physical activity has been related to reduced mortality, but it is not clear whether changes in physical activity affect mortality among older women. OBJECTIVE: To examine the relationship of changes in physical activity and mortality among older women. DESIGN, SETTING, AND PARTICIPANTS: Prospective cohort study conducted at 4 US research centers (Baltimore, Md; Portland, Ore; Minneapolis, Minn; and Monongahela Valley, Pa) among 9518 community-dwelling white women aged 65 years or older who were assessed at baseline (1986-1988), 7553 of whom were reassessed at a follow-up visit (1992-1994; median, 5.7 years later). MAIN OUTCOME MEASURES: Walking and other physical activities at baseline and follow-up; vital status, with cause of death confirmed by death certificates/discharge summaries, tracked for up to 12.5 years after baseline (up to 6.7 years after the follow-up visit). RESULTS: Compared with continually sedentary women, those who increased physical activity levels between baseline and follow-up had lower mortality from all causes (hazard rate ratio [HRR], 0.52; 95% confidence interval [CI], 0.40-0.69), cardiovascular disease (HRR, 0.64; 95% CI, 0.42-0.97), and cancer (HRR, 0.49; 95% CI, 0.29-0.84), independent of age, smoking, body mass index, comorbid conditions, and baseline physical activity level. Associations between changes in physical activity and reduced mortality were similar in women with and without chronic diseases but tended to be weaker among women aged at least 75 years and among those with poor health status. Women who were physically active at both visits also had lower all-cause mortality (HRR, 0.68; 95% CI, 0.56-0.82) and cardiovascular mortality (HRR, 0.62; 95% CI, 0.44-0.88) than sedentary women. CONCLUSION: Increasing and maintaining physical activity levels could lengthen life for older women but appears to provide less benefit for women aged at least 75 years and those with poor health status. PMID- 12746362 TI - Patterns of functional decline at the end of life. AB - CONTEXT: Clinicians have observed various patterns of functional decline at the end of life, but few empirical data have tested these patterns in large populations. OBJECTIVE: To determine if functional decline differs among 4 types of illness trajectories: sudden death, cancer death, death from organ failure, and frailty. DESIGN, SETTING, AND PARTICIPANTS: Cohort analysis of data from 4 US regions in the prospective, longitudinal Established Populations for Epidemiologic Studies of the Elderly (EPESE) study. Of the 14 456 participants aged 65 years or older who provided interviews at baseline (1981-1987), 4871 died during the first 6 years of follow-up; 4190 (86%) of these provided interviews within 1 year before dying. These decedents were evenly distributed in 12 cohorts based on the number of months between the final interview and death. MAIN OUTCOME MEASURES: Self- or proxy-reported physical function (performance of 7 activities of daily living [ADLs]) within 1 year prior to death; predicted ADL dependency prior to death. RESULTS: Mean function declined across the 12 cohorts, simulating individual decline in the final year of life. Sudden death decedents were highly functional even in the last month before death (mean [95% confidence interval [CI]] numbers of ADL dependencies: 0.69 [0.19-1.19] at 12 months before death vs 1.22 [0.59-1.85] at the final month of life, P =.20); cancer decedents were highly functional early in their final year but markedly more disabled 3 months prior to death (0.77 [0.30-1.24] vs 4.09 [3.37-4.81], P<.001); organ failure decedents experienced a fluctuating pattern of decline, with substantially poorer function during the last 3 months before death (2.10 [1.49-2.70] vs 3.66 [2.94 4.38], P<.001); and frail decedents were relatively more disabled in the final year and especially dependent during the last month (2.92 [2.24-3.60] vs 5.84 [5.33-6.35], P<.001). After controlling for age, sex, race, education, marital status, interval between final interview and death, and other demographic differences, frail decedents were more than 8 times more likely than sudden death decedents to be ADL dependent (OR, 8.32 [95% CI, 6.46-10.73); cancer decedents, one and a half times more likely (OR, 1.57 [95% CI, 1.25-1.96]); and organ failure decedents, 3 times more likely (OR, 3.00 [95% CI, 2.39-3.77]). CONCLUSIONS: Trajectories of functional decline at the end of life are quite variable. Differentiating among expected trajectories and related needs would help shape tailored strategies and better programs of care prior to death. PMID- 12746363 TI - Risk factors for pediatric human immunodeficiency virus-related malignancy. AB - CONTEXT: Although cancers occur with increased frequency in children with human immunodeficiency virus (HIV) infection, the specific clinical, immunological, and viral risk factors for malignancy have not been identified. OBJECTIVE: To identify risk factors for malignancy among HIV-infected children. DESIGN, SETTING, AND PATIENTS: A multicenter case-control study of children with HIV at 26 institutions participating in the Pediatric Oncology Group. Forty-three case patients with a new malignancy and 74 control patients without a malignancy were matched based on the duration of their infection. Patients were enrolled between January 1992 and July 1998. MAIN OUTCOME MEASURES: Clinical and laboratory factors assessed as putative risk factors included demographic characteristics, HIV characteristics, prior antiretroviral treatment, and CD4 cell count. Coviral infections with Epstein-Barr virus (EBV), cytomegalovirus, and human herpesvirus 6 were assessed by semiquantitative polymerase chain reaction assays and serological testing. RESULTS: Case malignancy diagnoses included 28 non-Hodgkin lymphoma, 4 B-cell acute lymphoblastic leukemia, 1 Hodgkin disease, 8 leiomyosarcoma, 1 hepatoblastoma, and 1 schwannoma. Epstein-Barr virus viral load of more than 50 viral genome copies per 105 peripheral blood mononuclear cells was strongly associated with cancer risk but only for children with CD4 cell counts of at least 200/ microL (odds ratio [OR], 11.33; 95% confidence interval [CI], 2.09-65.66, P<.001). High EBV viral load was not associated with cancer for children with CD4 cell counts of less than 200/ microL (OR, 1.12; 95% CI, 0.13 9.62; P =.99). Zidovudine antiretroviral therapy did not confer a significant protective effect for either the high (OR, 0.81; 95% CI, 0.22-3.09; P =.77) or the low CD4 cell count groups (OR, 0.27; 95% CI, 0.04-1.46; P =.16). The route of HIV infection was not associated with increased cancer risk. CONCLUSIONS: Route of infection, demographic characteristics, and zidovudine use were not associated with the development of malignancy in HIV-infected children. High viral burden with EBV was associated with the development of malignancy in HIV-infected children although the effect was modified by CD4 cell count. The pathogenesis of HIV-related pediatric malignancies remains unclear and other contributing risk factors can be elucidated only through further study. PMID- 12746364 TI - Use of the Internet and e-mail for health care information: results from a national survey. AB - CONTEXT: The Internet has attracted considerable attention as a means to improve health and health care delivery, but it is not clear how prevalent Internet use for health care really is or what impact it has on health care utilization. Available estimates of use and impact vary widely. Without accurate estimates of use and effects, it is difficult to focus policy discussions or design appropriate policy activities. OBJECTIVES: To measure the extent of Internet use for health care among a representative sample of the US population, to examine the prevalence of e-mail use for health care, and to examine the effects that Internet and e-mail use has on users' knowledge about health care matters and their use of the health care system. DESIGN, SETTING, AND PARTICIPANTS: Survey conducted in December 2001 and January 2002 among a sample drawn from a research panel of more than 60 000 US households developed and maintained by Knowledge Networks. Responses were analyzed from 4764 individuals aged 21 years or older who were self-reported Internet users. MAIN OUTCOME MEASURES: Self-reported rates in the past year of Internet and e-mail use to obtain information related to health, contact health care professionals, and obtain prescriptions; perceived effects of Internet and e-mail use on health care use. RESULTS: Approximately 40% of respondents with Internet access reported using the Internet to look for advice or information about health or health care in 2001. Six percent reported using e-mail to contact a physician or other health care professional. About one third of those using the Internet for health reported that using the Internet affected a decision about health or their health care, but very few reported impacts on measurable health care utilization; 94% said that Internet use had no effect on the number of physician visits they had and 93% said it had no effect on the number of telephone contacts. Five percent or less reported use of the Internet to obtain prescriptions or purchase pharmaceutical products. CONCLUSIONS: Although many people use the Internet for health information, use is not as common as is sometimes reported. Effects on actual health care utilization are also less substantial than some have claimed. Discussions of the role of the Internet in health care and the development of policies that might influence this role should not presume that use of the Internet for health information is universal or that the Internet strongly influences health care utilization. PMID- 12746365 TI - Effect of a practice-based strategy on test ordering performance of primary care physicians: a randomized trial. AB - CONTEXT: Numbers of diagnostic tests ordered by primary care physicians are growing and many of these tests seem to be unnecessary according to established, evidence-based guidelines. An innovative strategy that focused on clinical problems and associated tests was developed. OBJECTIVE: To determine the effects of a multifaceted strategy aimed at improving the performance of primary care physicians' test ordering. DESIGN: Multicenter, randomized controlled trial with a balanced, incomplete block design and randomization at group level. Thirteen groups of primary care physicians underwent the strategy for 3 clinical problems (arm A; cardiovascular topics, upper and lower abdominal complaints), while 13 other groups underwent the strategy for 3 other clinical problems (arm B; chronic obstructive pulmonary disease and asthma, general complaints, degenerative joint complaints). Each arm acted as a control for the other. SETTING: Primary care physician groups in 5 regions in the Netherlands with diagnostic centers recruited from May to September 1998. STUDY PARTICIPANTS: Twenty-six primary care physician groups, including 174 primary care physicians. INTERVENTION: During the 6 months of intervention, physicians discussed 3 consecutive, personal feedback reports in 3 small group meetings, related them to 3 evidence-based clinical guidelines, and made plans for change. MAIN OUTCOME MEASURE: According to existing national, evidence-based guidelines, a decrease in the total numbers of tests ordered per clinical problem, and of some defined inappropriate tests, is considered a quality improvement. RESULTS: For clinical problems allocated to arm A, the mean total number of requested tests per 6 months per physician was reduced from baseline to follow-up by 12% among physicians in the arm A intervention, but was unchanged in the arm B control, with a mean reduction of 67 more tests per physician per 6 months in arm A than in arm B (P =.01). For clinical problems allocated to arm B, the mean total number of requested tests per 6 months per physician was reduced from baseline to follow-up by 8% among physicians in the arm B intervention, and by 3% in the arm A control, with a mean reduction of 28 more tests per physician per 6 months in arm B than in arm A (P =.22). Physicians in arm A had a significant reduction in mean total number of inappropriate tests ordered for problems allocated to arm A, whereas the reduction in inappropriate test ordered physicians in arm B for problems allocated to arm B was not statistically significant. CONCLUSION: In this study, a practice-based, multifaceted strategy using guidelines, feedback, and social interaction resulted in modest improvements in test ordering by primary care physicians. PMID- 12746367 TI - Defending professional independence: ACCME's proposed new guidelines for commercial support of CME. PMID- 12746366 TI - Chronic hepatitis C virus infection. PMID- 12746368 TI - Hypertension prevalence and stroke mortality across populations. PMID- 12746369 TI - Alcohol advertising in magazines and adolescent readership. AB - CONTEXT: Adolescent drinking is a major public health concern. The federal government does not restrict alcohol advertising to adolescents, but relies on the alcohol industry for self-regulation. OBJECTIVES: To investigate recent alcohol advertising in magazines and to determine whether advertising frequency is associated with adolescent readership. DESIGN, SETTING, AND SUBJECTS: All alcohol advertisements were counted that appeared from 1997-2001 in 35 of 48 major US magazines, which tracked their adolescent readership (3 refused all alcohol advertisements; and advertisement counts were unavailable for 10). Variation was assessed in the advertisement placement frequency for each major category of alcohol (beer, wine and wine coolers, and distilled liquor) by a magazine's adolescent readership (age 12-19 years), young adult readership (age 20-24 years), and older adult readership (age > or =25 years); readership demographics (sex, race, and income); year; frequency of publication; and cost per advertisement. MAIN OUTCOME MEASURE: Variation in alcohol advertising frequency by adolescent readership. RESULTS: Adolescent readership ranged from 1.0 to 7.1 million. The alcohol industry placed 9148 advertisements at a cost of 696 million dollars. Of the 9148 advertisements, 1201 (13%) were for beer, 443 (5%) for wine, and 7504 (82%) for liquor. After adjustment for other magazine characteristics, the advertisement rate ratio was 1.6 times more for beer (95% confidence interval [CI], 1.0-2.6; P =.05) and liquor (95% CI, 1.1-2.3; P =.01) for every additional million adolescent readers. Wine industry advertising was not associated with adolescent readership. CONCLUSIONS: Magazine advertising by the beer and liquor industries is associated with adolescent readership. Industry and federal policymakers should examine ways to regulate advertising that reaches large numbers of adolescents. PMID- 12746370 TI - JAMA patient page. Hepatitis C. PMID- 12746371 TI - Genetic linkage of the penicillinase gene, amp, and blrAB, encoding the regulator of beta-lactamase expression in Aeromonas spp. AB - Aeromonas hydrophila T429125, a human clinical isolate, possesses three coordinately inducible beta-lactamases encoded by ampH (class D beta-lactamase), cepH (class C beta-lactamase) and imiH (class B beta-lactamase). We report that upstream of ampH there are two genes, blrA and blrB, encoding a putative two component regulatory system. PCR studies revealed the same blrAB-amp gene arrangement in all Aeromonas spp. isolates tested; namely, Aeromonas veronii bv. sobria, Aeromonas jandaei, Aeromonas mediae, Aeromonas salmonicida and Aeromonas trota. A dominant mutation in the predicted BlrB kinase domain results in beta lactamase overexpression in A. hydrophila T429125, but in other beta-lactamase overexpressing mutants blrAB remains intact. Relative to the parent strain, A. hydrophila T429125, beta-lactamase- overexpressing mutants show a clear hierarchy of increased beta-lactamase expression: ImiH > CepH > AmpH. The same hierarchy is seen following beta-lactam challenge of A. hydrophila T429125, and correlates with the number of blr-tag sequences (TTCAC) found upstream of each beta lactamase gene: ampH (one), cepH (two) and imiH (three). PMID- 12746372 TI - Antibiotics and hospital-acquired Clostridium difficile-associated diarrhoea: a systematic review. AB - A systematic review of studies that investigated the association of antibiotics with hospital-acquired Clostridium difficile-associated diarrhoea (CDAD) was undertaken to summarize the strength of the evidence for this relationship. The results from the studies identified were considered after critically reviewing the design and conduct of each study. Although the majority of studies found an association with various antibiotics, antibiotic classes or components of antibiotic administration, most were limited in their ability to establish a causal relationship by the use of incorrect control groups, the presence of bias, inadequate control of confounding and small sample sizes. The limitations identified in this review prevented the pooling of results in a meta-analysis. Two studies of reasonable quality suggested an association between clindamycin, cephalosporins, penicillins and CDAD. Well-designed studies grounded in epidemiological principles are needed to identify true risk factors for CDAD and to provide reliable estimates of the strength of association. PMID- 12746373 TI - Effects of antimicrobial therapy on the microbial flora of the adenoids. AB - The core of the adenoids contains polymicrobial aerobic and anaerobic flora and also includes potential respiratory pathogens. Similar flora, although in higher numbers and with a higher frequency of pathogens, are found in inflamed or hypertrophic adenoids and many of these bacteria are resistant to antimicrobial agents. Exposure to antimicrobial therapy can alter the colonization patterns and select for resistant organisms. Production of beta-lactamase is one of the major mechanisms of resistance of these organisms. The adenoids of healthy individuals, in contrast to those with recurrent respiratory tract infections, are generally colonized by aerobic and anaerobic organisms that are capable of interfering with the growth of potential pathogens. Maintaining the beneficial effects of normal flora by avoiding unnecessary exposure to antimicrobial therapy may be a useful tool in preventing colonization of the adenoids by potential pathogens. This review discusses the unique microbiology of the adenoids in individuals with a variety of pathological conditions, the interactions between the various organisms and the effects of antimicrobial therapy on the microbial flora of the adenoids. PMID- 12746374 TI - Improving the mouse model for studying the efficacy of voriconazole. AB - Outbred ICR mice were rendered neutropenic, infected intravenously with Fusarium solani and treated orally with voriconazole. When given alone, voriconazole was not protective up to 40 mg/kg/day. When grapefruit juice was administered before infection, mice were protected by voriconazole. The mechanism may be inhibition of gut mucosal cytochrome enzymes that rapidly degrade voriconazole in the mouse. These murine studies support expansion of voriconazole therapy in other highly resistant systemic mycoses. PMID- 12746376 TI - In vitro activity of faropenem compared with eight agents against fourteen Gram positive and Gram-negative bacteria by time-kill. PMID- 12746375 TI - Intrapulmonary penetration of linezolid. AB - OBJECTIVE: This study was designed to measure the concentrations of linezolid in bronchial mucosa, pulmonary macrophages and epithelial lining fluid and to compare them with simultaneous blood levels. METHODS: Ten adult patients undergoing bronchoscopy for diagnostic purposes were given oral linezolid at a dosage of 600 mg twice a day for a total of six doses. Patients with active lung infection were excluded from the study. Flexible bronchoscopy was carried out between 2 and 8 h after the last dose of linezolid. Bronchial biopsies and bronchoalveolar lavage were carried out and a simultaneous blood sample obtained. Linezolid levels were measured using high-performance liquid chromatography (HPLC). RESULTS: Mean concentrations of linezolid were 13.4 mg/L in serum, 10.7 mg/kg in mucosa, 8.1 mg/L in alveolar macrophages and 25.1 mg/L in epithelial lining fluid. The mean site/serum concentration ratios were 0.79 for bronchial mucosa, 0.71 for macrophages and 8.35 for epithelial lining fluid. CONCLUSIONS: The MIC90 (< or =4 mg/L) of linezolid for Staphylococcus aureus and Streptococcus pneumoniae was exceeded in serum and bronchial mucosa in all subjects, in epithelial lining fluid in nine subjects and in macrophages in six subjects. PMID- 12746378 TI - Bacteraemia due to Bacteroides fragilis with reduced susceptibility to metronidazole. PMID- 12746379 TI - Health promotion in Africa: strategies, players, challenges and prospects. PMID- 12746377 TI - Adherence to local hospital guidelines for surgical antimicrobial prophylaxis: a multicentre audit in Dutch hospitals. AB - OBJECTIVE: To study the adherence to local hospital guidelines for antimicrobial prophylaxis in surgery, and explore reasons for non-adherence. METHODS: A prospective, multicentre audit of elective procedures, without prior suspicion of infection, was carried out in 13 Dutch hospitals. By reviewing medical, anaesthetic and nursing records, and medication charts, the prescription of antibiotics was compared with the local hospital guideline on antibiotic choice, duration of prophylaxis, dose, dosing interval and timing of the first dose. RESULTS: Between January 2000 and January 2001, 1763 procedures were studied. Antibiotic choice, duration, dose, dosing interval and timing of the first dose were concordant with the hospital guideline in 92%, 82%, 89%, 43% and 50%, respectively. Overall adherence to all aspects of the guideline, however, was achieved in only 28%. The most important barriers to local guideline adherence were lack of awareness due to ineffective distribution of the most recent version of the guidelines, lack of agreement by surgeons with the local hospital guidelines, and environmental factors, such as organizational constraints in the surgical suite and in the ward. CONCLUSION: This study shows that, although adherence to separate aspects of local hospital guidelines for surgical prophylaxis in the Netherlands is favourable, overall adherence to all parameters is hard to achieve. Adherence to guidelines on dosing interval and timing needs improvement, in particular. To increase the quality of antimicrobial prophylaxis in surgery, effort should be put into developing guidelines acceptable to surgeons, in adequately distributing the guidelines and to facilitating logistics. Audits of surgical prophylaxis may help hospitals identify barriers to guideline adherence. PMID- 12746380 TI - The value of community participation in disease surveillance: a case study from Niger. AB - A team of researchers, including one behavioral scientist (S.M.N.) and three epidemiologists (L.Q., O.S. and S.N.) conducted community analyses to assess the social and cultural factors that affect the detection and reporting of disease cases in a surveillance system, using acute flaccid paralysis (AFP) surveillance in Niger as a case study. Over a 60-day period in the country, the research team reviewed written field reports and interviewed epidemiologists, nurses, community members and persons in governmental and non-governmental organizations. Overall, we found that the logistical difficulties of travel and communication, which are common in developing countries, constrain the conventional surveillance system that relies on epidemiologists visiting sites to discover and investigate cases, particularly in rural areas. Other challenges include: community members' lack of knowledge about the possible link between a case of paralysis and a dangerous, communicable disease; lack of access to health care, including the low number of clinics and health care workers; cultural beliefs that favor seeking a local healer before consulting a nurse or physician; and health workers' lack of training in AFP surveillance. The quality of surveillance in developing countries can improve if a community-based approach is adopted. Such a system has been used successfully in Niger during smallpox-eradication and guinea worm-control campaigns. In a community-based system, community members receive basic education or more extensive training to motivate and enable them to notify health care staff about possible cases of disease in a timely fashion. Local organizations, local projects and local leaders must be included to ensure the success of such a program. In Niger we found sufficient quantities of this type of social capital, along with enough local experience of past health campaigns, to suggest that a community-based approach can improve the level of comprehensiveness and sensitivity of surveillance. PMID- 12746382 TI - Beijing health promoting universities: practice and evaluation. AB - The aims of this study were to create a health promoting university within the framework of the Ottawa Charter for Health Promotion. Strategies included reforming and issuing healthy policies, creating a healthy physical and social environment, developing personal health skills, reorienting the health services, and implementing intervention activities. To evaluate the study, 180 students and 120 teaching/administrative staff were sampled for an in-depth interview with open-ended questions administered 1 year after the launch of the project. To assess health knowledge and behavior, 2500 students were sampled to answer a questionnaire, both prior to and following project implementation. With respect to policies, environment and health services, 166 students and 117 teaching and administrative staff participated in the in-depth interview. Approximately three quarters (75.90%) of university students considered that the physical environment of the campus had improved significantly and 83.73% reported they had a good social environment. All university administration departments made commitments to health promotion. Consultations on mental health, smoking cessation and STD/AIDS prevention were provided all year round. Health education was included in a curriculum as a selective course with 1-2 credits. Almost two-thirds (60.66%) of teaching/administrative staff reported that they had had a yearly physical examination. In the final stages of the research, significantly more college students reported improved mental health (38.25% compared with 17.93% at baseline) (p < 0.01) and more were knowledgeable about transmission of STDs/AIDS (57.00/35.50% compared with 51.66/28.20% at baseline, respectively) (p < 0.01). Significantly less regular smokers were found (45% compared with 15.81% at baseline) (p < 0.01). However, there was a significant increase in high-fat food intake (44.81% compared with 49.50%) (p < 0.01) and pre-marital sex (5.11% compared with 14.00%), and a significant decrease in physical exercise participation (29.41% compared with 23.50%) (p < 0.01). As a health promotion setting, the university community can benefit greatly from implementing health promotion campaigns based on the principles of the Ottawa Charter. PMID- 12746381 TI - Building capable communities: experiences in a rural Fijian context. AB - This paper discusses the design of a methodology for 'building capable communities' in a health promotion programme context. The design of the methodology builds upon previous work and offers a new approach, through the use of nine 'operational domains', for the assessment and strategic planning of community empowerment. The purpose is to go beyond the rhetoric of participation and empowerment, and to provide a better understanding of how community empowerment goals can be made to be operational in a health promotion programme context. The experiences of implementing the methodology in two rural Fijian communities are discussed briefly. The common themes for the successful application of the methodology have been identified. The paper will be of interest to the planners and evaluators of health promotion programmes that aim to build community capacity and promote empowerment. PMID- 12746384 TI - Evaluating the Fabreville Heart Health Program in Laval, Canada: a dialogue between two paradigms, positivism and constructivism. AB - As part of the Canadian Federal-Provincial Initiative in Heart Health, the goal of the Fabreville Heart Health Program was to sensitize a district of Laval, Quebec's second most populous city, to heart-healthy behaviours. The program was planned and implemented by a committee composed of Fabreville community leaders and professionals from the Public Health Department. Between 1992 and 1994, intervention objectives were defined by the department in terms of changing individual behaviours associated with cardiovascular risk factors, namely diet, sedentariness and smoking, as well as adapting physical and social environments to facilitate these changes. However, from 1994 to its conclusion in 1997, the program was re-oriented to engage the population in mobilizing their own community and taking charge of interventions themselves. Actions then became dependent on the interests and motivation of Fabreville residents to transform their lifestyles and aspects of their physical environment. The initial evaluation process, based on the positivist paradigm, was designed to measure changes in individual behaviours and certain physical environments, such as an increase in designated non-smoking areas. However, following the re-orientation towards community mobilization, it was decided that evaluation should go beyond the professional production of data to include a process of the collective construction of knowledge. Evaluation methodology then became based on the constructivist paradigm. Yet field constraints such as lack of community involvement in both leadership and process evaluation, and the need to ensure evaluation standards and fulfil sponsor obligations, compelled the Public Health Department to return to using a certain number of positivist methods. The ensuing inter-paradigm dialogue helped broaden the scope of evaluation and contributed to gaining a more in-depth understanding of the processes and outcomes of community mobilization. PMID- 12746383 TI - Social context for workplace health promotion: feasibility considerations in Costa Rica, Finland, Germany, Spain and Sweden. AB - We constructed a simple, flexible procedure that facilitates the pre-assessment of feasibility of workplace health promotion (WHP) programmes. It evaluates cancer hazards, workers' need for hazard reduction, acceptability of WHP, and social context. It was tested and applied in 16 workplace communities and among 1085 employees in industry, construction, transport, services, teaching and municipal works in Costa Rica, Finland, Germany, Spain and Sweden. Social context is inseparable from WHP. It covers workers' organizations and representatives, management, safety committees, occupational health services, health and safety enforcement agencies, general health services, non-government organizations, insurance systems, academic and other institutions, regulatory stipulations pertaining WHP, and material resources. Priorities, risk definitions, attitudes, hazard profiles, motivations and assessment methods were highly contextual. Management preferred passive interventions, helping cover expert costs, participating in planning and granting time. Trade unions, workers' representatives, safety committees and occupational health services appeared to be important operational partners. Occupational health services may however be loaded with curative and screening functions or be non-existent. We advocate participatory, multifaceted WHP based on the needs and empowerment of the workers themselves, integrating occupational and lifestyle hazards. Workforce in irregular and shift work, in agriculture, in small enterprises, in the informal sector, and immigrant, seasonal and temporary workers represent groups in need of particular strategies such as community health promotion. In a more general framework, social context itself may become a target for intervention. PMID- 12746385 TI - Factors in creating sustainable intersectoral community mobilization for prevention of heart and lung disease. AB - This paper describes factors facilitating and working against successful community mobilization in the implementation of an integrated prevention programme for cardiovascular disease and lung cancer in four community settings in Quebec, Canada. Implementation evaluation data from several sources showed that over the 3-year period, mobilization was partly achieved in all four communities, although the degree of success varied. The data support those of previous studies showing that several factors are key to effective intersectoral community mobilization: (i) involvement of concerned and influential community members with a commitment to shared goals and a visible community focus; (ii) formation of multi-organization systems among appropriate organizations, recognizing their strengths, resources and competencies, and preserving both their autonomy and interdependence with an appreciation of divergent perspectives; (iii) development of decision-making mechanisms through the setting up of formal structural arrangements to facilitate decisions with clear leadership; (iv) clear definition of objectives, tasks, roles and responsibilities; and (v) official support and legitimization from participating agencies, government authorities, and organizations with adequate resources devoted to partnership building. This study also replicated a number of barriers to the creation of sustainable intersectoral community mobilization, notably the potentially destructive role of power conflicts among the key institutional partners. PMID- 12746386 TI - Critical processes for creating health-promoting sporting environments in Australia. AB - The reach of sporting organizations into the community makes them an ideal vehicle through which to promote health to the general population. There are now a number of documented examples demonstrating that sponsorship can lead to improvements in the health of the sporting environment, but relatively little is known as to why some sponsorships are more successful in achieving these structural changes than others in ostensibly similar sports. The purpose of this study was to identify the processes required for health promotion agencies and sporting organizations working in collaboration to implement structural changes in sporting settings such as smoke-free environments, provision of healthy food choices, responsible alcohol management and sun protection, along with the factors that facilitate and hinder this from being achieved. We conclude that such changes are difficult to achieve, especially in the absence of a programmatic approach to health promotion. PMID- 12746387 TI - A cross-cultural analysis of 'motivation for eating' as a potential factor in the emergence of global obesity: Japan and the United States. AB - This exploratory study compared motivation for eating between individuals from two different cultures that have moved through the nutrition transition at different rates and to different degrees. The analysis was based on a convenience sample of 1218 participants aged >or=18 years attending colleges in the US and Japan. The Motivation for Eating Scale (MFES) was used to evaluate different motivations for eating by nation and gender. The MFES consists of 12 items classified into three subscales: emotional, physical and environmental eating. The questionnaire used in the study also included responses about participants' motivation to lose weight, frequency of dieting, presence of previous or existing eating disorders, and frequency of exercise. Results showed no significant differences in the three MFES subscales for men in the US and Japan. For women, however, significant differences were seen for all three subscales. Women in the US were more likely to initiate eating for emotional reasons, while women in Japan were more likely to eat for physical or environmental reasons. Women and men in the US were more likely than the Japanese respondents to eat in response to watching TV or movies. These results suggest that there are national differences in the cultural environment that may impact individual motivations for eating. As such, various cultural perceptions of food should be considered in attempts to understand more fully the mechanics of the nutrition transition as it operates within a given country. By extension, public health policies and health promotion initiatives that are designed to limit the negative impacts of the nutrition transition may benefit from a greater understanding of the larger role that cultural perceptions of food may play in influencing individual motivations for eating. PMID- 12746388 TI - The 'wonderfulness' of children's feeding programs. AB - When people involved in children's feeding programs were asked to describe them, without exception they were described using phrases that reflected the perception of 'wonderfulness'. This paper critically analyses the 'wonderfulness' of children's feeding programs by examining the language used to describe these programs, and the features of a 'wonderful' program through an analysis of a multi-site, qualitative case study of nine diverse programs in Atlantic Canada. When participants justified their comments about the 'wonderfulness' of children's feeding programs, they did so based upon five perceptions of program strengths: enhanced family coping; providing good food and nutrition; socializing and making friends; behaving well in school; and volunteerism. We suggest that programs can be designed to be innately 'wonderful' if they are community- and charity-based, support a noble cause such as the elimination of child hunger, engage good people as donors and volunteers, and provide a direct service to children apart from their families. We challenge health promoters to beware of the 'wonderful' program; its 'wonderfulness' may actually be masking unintended negative impacts upon its participants. PMID- 12746389 TI - IUHPE partnership in European networks and projects. PMID- 12746390 TI - Mowat-Wilson syndrome. AB - MWS is a multiple congenital anomaly syndrome, first clinically delineated by Mowat et al in 1998. Over 45 cases have now been reported. All patients have typical dysmorphic features in association with severe intellectual disability, and nearly all have microcephaly and seizures. Congenital anomalies, including Hirschsprung disease (HSCR), congenital heart disease, hypospadias, genitourinary anomalies, agenesis of the corpus callosum, and short stature are common. The syndrome is the result of heterozygous deletions or truncating mutations of the ZFHX1B (SIP1) gene on chromosome 2q22. PMID- 12746391 TI - Molecular pathology and genetics of congenital hepatorenal fibrocystic syndromes. AB - The hepatorenal fibrocystic (HRFC) syndromes are a heterogeneous group of severe monogenic conditions that may be detected before birth. Commonly, HRFC syndromes present in the neonatal and paediatric age, with consistent developmental abnormalities mostly involving the liver and kidney. The changes include the proliferation and dilatation of epithelial ducts in these tissues with abnormal deposition of extracellular matrix. In this review, we examine the clinical features and differential diagnoses of this group of syndromes, including autosomal recessive polycystic kidney disease (ARPKD), juvenile nephronophthisis (NPHP), Meckel-Gruber syndrome (MKS), Bardet-Biedl syndrome (BBS), and Jeune asphyxiating thoracic dystrophy (JATD). Extrahepatic manifestations include mostly bone and central nervous system abnormalities, dysmorphic features, and developmental delay. Previously, it has been suggested that ARPKD, JATD, and Ellis-van Creveld syndrome (EvC) may arise from defects in differentiation in a common developmental pathway. We review recent molecular advances in the recessive HRFC syndromes and discuss this hypothesis. PMID- 12746393 TI - Disruption of the neuronal PAS3 gene in a family affected with schizophrenia. AB - Schizophrenia and its subtypes are part of a complex brain disorder with multiple postulated aetiologies. There is evidence that this common disease is genetically heterogeneous, with many loci involved. In this report, we describe a mother and daughter affected with schizophrenia, who are carriers of a t(9;14)(q34;q13) chromosome. By mapping on flow sorted aberrant chromosomes isolated from lymphoblast cell lines, both subjects were found to have a translocation breakpoint junction between the markers D14S730 and D14S70, a 683 kb interval on chromosome 14q13. This interval was found to contain the neuronal PAS3 gene (NPAS3), by annotating the genomic sequence for ESTs and performing RACE and cDNA library screenings. The NPAS3 gene was characterised with respect to the genomic structure, human expression profile, and protein cellular localisation to gain insight into gene function. The translocation breakpoint junction lies within the third intron of NPAS3, resulting in the disruption of the coding potential. The fact that the bHLH and PAS domains are disrupted from the remaining parts of the encoded protein suggests that the DNA binding and dimerisation functions of this protein are destroyed. The daughter (proband), who is more severely affected, has an additional microdeletion in the second intron of NPAS3. On chromosome 9q34, the translocation breakpoint junction was defined between D9S752 and D9S972 and no genes were found to be disrupted. We propose that haploinsufficiency of NPAS3 contributes to the cause of mental illness in this family. PMID- 12746392 TI - Canine tricuspid valve malformation, a model of human Ebstein anomaly, maps to dog chromosome 9. AB - BACKGROUND: Ebstein anomaly of the tricuspid valve is a congenital cardiac malformation characterised by downward displacement of the attachment of the septal and posterior leaflets of the tricuspid valve. Canine tricuspid valve malformation (CTVM) is morphologically similar to Ebstein anomaly; familial occurrence of CTVM has been described. Several observations suggest a genetic cause but most cases appear to be sporadic. METHODS: Three purebred Labrador Retriever kindreds enriched for CTVM underwent clinical examination and echocardiography. DNA was extracted from whole blood. Genotyping was carried out using polymorphic repeat markers with an average spacing of 15 cM and polymorphic information content of 0.74. RESULTS: Pedigree analysis identified CTVM segregating as an autosomal dominant trait with reduced penetrance. Genome wide linkage analysis in one kindred identified a CTVM susceptibility locus on dog chromosome 9 (CFA9) with a maximum multipoint lod score of 3.33. The two additional kindreds showed a conserved disease haplotype. CONCLUSIONS: This study identifies a CTVM susceptibility locus on CFA9 and a founder effect in apparently unrelated Labrador Retriever kindreds. These results provide the basis for a positional candidate cloning effort to identify the CTVM disease gene. Identification of the CTVM gene will permit mutation screening of patients with Ebstein anomaly, which should provide additional insights into the genetic programmes of valve development. PMID- 12746395 TI - A genome scan for developmental dyslexia confirms linkage to chromosome 2p11 and suggests a new locus on 7q32. AB - Developmental dyslexia is a distinct learning disability with unexpected difficulty in learning to read despite adequate intelligence, education, and environment, and normal senses. The genetic aetiology of dyslexia is heterogeneous and loci on chromosomes 2, 3, 6, 15, and 18 have been repeatedly linked to it. We have conducted a genome scan with 376 markers in 11 families with 38 dyslexic subjects ascertained in Finland. Linkage of dyslexia to the vicinity of DYX3 on 2p was confirmed with a non-parametric linkage (NPL) score of 2.55 and a lod score of 3.01 for a dominant model, and a novel locus on 7q32 close to the SPCH1 locus was suggested with an NPL score of 2.77. The SPCH1 locus has previously been linked with a severe speech and language disorder and autism, and a mutation in exon 14 of the FOXP2 gene on 7q32 has been identified in one large pedigree. Because the language disorder associated with the SPCH1 locus has some overlap with the language deficits observed in dyslexia, we sequenced the coding region of FOXP2 as a candidate gene for our observed linkage in six dyslexic subjects. No mutations were identified. We conclude that DYX3 appears to be important for dyslexia susceptibility in many Finnish families, and a suggested linkage of dyslexia to chromosome 7q32 will need verification in other data sets. PMID- 12746396 TI - Application of haplotype pair analysis for the identification of hemizygous loci. AB - An expectation maximisation based prediction algorithm was created to identify unusual haplotypes in patient samples that may be caused by small intragenic deletions. In this approach, unphased SNP genotypes are compared to pairs of canonical haplotypes to identify potentially hemizygous regions. This method was successfully applied to identify five deletions in the 3' region of BRCA1. PMID- 12746394 TI - Novel mutations in DLL3, a somitogenesis gene encoding a ligand for the Notch signalling pathway, cause a consistent pattern of abnormal vertebral segmentation in spondylocostal dysostosis. AB - The spondylocostal dysostoses (SCD) are a group of disorders characterised by multiple vertebral segmentation defects and rib anomalies. SCD can either be sporadic or familial, and can be inherited in either autosomal dominant or recessive modes. We have previously shown that recessive forms of SCD can be caused by mutations in the delta-like 3 gene, DLL3. Here, we have sequenced DLL3 in a series of SCD cases and identified 12 mutations in a further 10 families. These include 10 novel mutations in exons 4-8, comprising nonsense, missense, frameshift, splicing, and in frame insertion mutations that are predicted to result in either the truncation of the mature protein in the extracellular domain, or affect highly conserved amino acid residues in the epidermal growth factor-like repeats of the protein. The affected cases represent diverse ethnic backgrounds and six come from traditionally consanguineous communities. In all affected subjects, the radiological phenotype is abnormal segmentation throughout the entire vertebral column with smooth outlines to the vertebral bodies in childhood, for which we suggest the term "pebble beach sign". This is a very consistent phenotype-genotype correlation and we suggest the designation SCD type 1 for the AR form caused by mutations in the DLL3 gene. PMID- 12746397 TI - Does apolipoprotein E polymorphism influence susceptibility to malaria? AB - Outcome of infection varies greatly among people, and in the case of three very different viruses, it is determined by apolipoprotein E (APOE) genotype. APOE might affect outcome of malaria infection also, since apoE protein and the protozoon (like the viruses) share cell entry mediators (heparan sulphate proteoglycans and/or specific apoE receptors). APOE polymorphisms give rise to protein variants that differ in binding strength to these mediators; thus, the extent of competition between apoE and protozoon for cell entry, and hence magnitude of protozoan damage, might depend on apoE isoform. Genotypes of infants infected with malaria were examined. It was found that APOE epsilon 2 homozygotes became infected at an earlier age than those carrying the other genotypes, the difference being statistically significant. Parasite densities, all of which were low, did not differ significantly. This effect, although based on small numbers, suggests that APOE epsilon 2 may be a risk factor for early infection. PMID- 12746398 TI - The neurobeachin gene is disrupted by a translocation in a patient with idiopathic autism. PMID- 12746399 TI - Rare polymorphic variants of the AGTR2 gene in boys with non-specific mental retardation. PMID- 12746401 TI - Mutations of the Birt-Hogg-Dube (BHD) gene in sporadic colorectal carcinomas and colorectal carcinoma cell lines with microsatellite instability. PMID- 12746402 TI - NF1 mutations and clinical spectrum in patients with spinal neurofibromas. PMID- 12746400 TI - Distinctive audiometric profile associated with DFNB21 alleles of TECTA. PMID- 12746403 TI - Gender specific association of aldosterone synthase gene polymorphism with renal survival in patients with IgA nephropathy. PMID- 12746404 TI - Predictive testing for cognitive functioning in female carriers of the fragile X syndrome using hair root analysis. PMID- 12746405 TI - Mutation analysis in the MECP2 gene and genetic counselling for Rett syndrome. PMID- 12746406 TI - Patients with the R133C mutation: is their phenotype different from patients with Rett syndrome with other mutations? PMID- 12746407 TI - Mapping of a de novo unequal crossover causing a deletion of the steroid 21 hydroxylase (CYP21A2) gene and a non-functional hybrid tenascin-X (TNXB) gene. PMID- 12746409 TI - Experience of discharge from colonoscopy of mutation negative HNPCC family members. PMID- 12746408 TI - Cancer surveillance is often inadequate in people at high risk for colorectal cancer. PMID- 12746410 TI - Women's preferences and consultants' communication of risk in consultations about familial breast cancer: impact on patient outcomes. PMID- 12746411 TI - Evidence for linkage and association of the markers near the LPL gene with hypertension in Chinese families. PMID- 12746412 TI - Haemochromatosis (HFE) gene C282Y mutation and the risk of coronary artery disease and myocardial infarction: a study in 1279 patients undergoing coronary angiography. PMID- 12746413 TI - Comparison of fluorescent SSCP and denaturing HPLC analysis with direct sequencing for mutation screening in hypertrophic cardiomyopathy. PMID- 12746414 TI - Multicolour FISH fine mapping unravels an insertion as a complex chromosomal rearrangement involving six breakpoints and a 5.89 Mb large deletion. PMID- 12746415 TI - Partial trisomy 10q with mild phenotype caused by an unbalanced X;10 translocation. PMID- 12746417 TI - Death in adults with Prader-Willi syndrome may be correlated with maternal uniparental disomy. PMID- 12746416 TI - Partial trisomy of chromosome 22 resulting from an interstitial duplication of 22q11.2 in a child with typical cat eye syndrome. PMID- 12746418 TI - Maternal 677CT/1298AC genotype of the MTHFR gene as a risk factor for cleft lip. PMID- 12746419 TI - Searching for genomic variants in the MESTIT1 transcript in Silver-Russell syndrome patients. PMID- 12746420 TI - A common IL-1 complex haplotype is associated with an increased risk of atopy. PMID- 12746421 TI - Novel sarcoglycan gene mutations in a large cohort of Italian patients. PMID- 12746422 TI - Contribution of connexin26 (GJB2) mutations and founder effect to non-syndromic hearing loss in India. PMID- 12746423 TI - Neuroferritinopathy in a French family with late onset dominant dystonia. PMID- 12746424 TI - ABCB4 gene sequence variation in women with intrahepatic cholestasis of pregnancy. PMID- 12746425 TI - A novel mutation (R218Q) at the boundary between the N-terminal and the first transmembrane domain of the glycine receptor in a case of sporadic hyperekplexia. PMID- 12746426 TI - Mutational and gross deletion study of the MEN1 gene and correlation with clinical features in Spanish patients. PMID- 12746427 TI - Mutation and LOH analysis of ACO2 in colorectal cancer: no evidence of biallelic genetic inactivation. PMID- 12746428 TI - The metal specificity and selectivity of ZntA from Escherichia coli using the acylphosphate intermediate. AB - ZntA from Escherichia coli is a P-type ATPase that confers resistance to Pb(II), Zn(II), and Cd(II) in vivo. We had previously shown that purified ZntA shows ATP hydrolysis activity with the metal ions Pb(II), Zn(II), and Cd(II). In this study, we utilized the acylphosphate formation activity of ZntA to further investigate the substrate specificity of ZntA. The site of phosphorylation was Asp-436, as expected from sequence alignments. We show that in addition to Pb(II), Zn(II), and Cd(II), ZntA is active with Ni(II), Co(II), and Cu(II), but not with Cu(I) and Ag(I). Thus, ZntA is specific for a broad range of divalent soft metal ions. The activities with Ni(II), Co(II), and Cu(II) are extremely low; the activities with these non-physiological substrates are 10-20-fold lower compared with the values obtained with Pb(II), Zn(II), and Cd(II). Similar results were obtained with DeltaN-ZntA, a ZntA derivative lacking the amino terminal metal binding domain. By characterizing the acylphosphate formation reaction in ZntA in detail, we show that a step prior to enzyme phosphorylation, most likely the metal ion binding step, is the slow step in the reaction mechanism in ZntA. The low activities with Ni(II), Co(II), and Cu(II) are because of a further decrease in the rate of binding of these metal ions. Thus, metal ion selectivity in ZntA and possibly other P1-type ATPases is based on the charge and the ligand preference of particular metal ions but not on their size. PMID- 12746429 TI - Characterization of Nkx3.2 DNA binding specificity and its requirement for somitic chondrogenesis. AB - We have previously shown that Nkx3.2, a member of the NK class of homeoproteins, functions as a transcriptional repressor to promote somitic chondrogenesis. However, it has not been addressed whether Nkx3.2 can bind to DNA in a sequence specific manner and whether DNA binding by Nkx3.2 is required for its biological activity. In this work, we employed a DNA binding site selection assay, which identified TAAGTG as a high affinity Nkx3.2 binding sequence. Sequence-specific binding of Nkx3.2 to the TAAGTG motif in vitro was confirmed by electrophoretic mobility shift assays, and mutagenesis of this sequence revealed that HRAGTG (where H represents A, C, or T, and R represents A or G) comprises the consensus DNA binding site for Nkx3.2. Consistent with these findings, the expression of a reporter gene containing reiterated Nkx3.2 binding sites was repressed in vivo by Nkx3.2 co-expression. In addition, we have generated a DNA nonbinding point mutant of Nkx3.2 (Nkx3.2-N200Q), which contains an asparagine to glutamine missense mutation in the homeodomain. Interestingly, despite being defective in DNA binding, Nkx3.2-N200Q still retains its intrinsic transcriptional repressor function. Finally, we demonstrate that unlike wild-type Nkx3.2, Nkx3.2-N200Q is unable to activate the chondrocyte differentiation program in somitic mesoderm, indicating that DNA binding by Nkx3.2 is critical for this factor to induce somitic chondrogenesis. PMID- 12746431 TI - A novel Ca2+-induced Ca2+ release mechanism in A7r5 cells regulated by calmodulin like proteins. AB - Intracellular Ca2+ release is involved in setting up Ca2+ signals in all eukaryotic cells. Here we report that an increase in free Ca2+ concentration triggered the release of up to 41 +/- 3% of the intracellular Ca2+ stores in permeabilized A7r5 (embryonic rat aorta) cells with an EC50 of 700 nm. This type of Ca2+-induced Ca2+ release (CICR) was neither mediated by inositol 1,4,5 trisphosphate receptors nor by ryanodine receptors, because it was not blocked by heparin, 2-aminoethoxydiphenyl borate, xestospongin C, ruthenium red, or ryanodine. ATP dose-dependently stimulated the CICR mechanism, whereas 10 mm MgCl2 abolished it. CICR was not affected by exogenously added calmodulin (CaM), but CaM1234, a Ca2+-insensitive CaM mutant, strongly inhibited the CICR mechanism. Other proteins of the CaM-like neuronal Ca2+-sensor protein family such as Ca2+-binding protein 1 and neuronal Ca2+ sensor-1 were equally potent for inhibiting the CICR. Removal of endogenous CaM, using a CaM-binding peptide derived from the ryanodine receptor type-1 (amino acids 3614-3643) prevented subsequent activation of the CICR mechanism. A similar CICR mechanism was also found in 16HBE14o-(human bronchial mucosa) cells. We conclude that A7r5 and 16HBE14o-cells express a novel type of CICR mechanism that is silent in normal resting conditions due to inhibition by CaM but becomes activated by a Ca2+ dependent dissociation of CaM. This CICR mechanism, which may be regulated by members of the family of neuronal Ca2+-sensor proteins, may provide an additional route for Ca2+ release that could allow amplification of small Ca2+ signals. PMID- 12746430 TI - Sphingosine 1-phosphate, a diffusible calcium influx factor mediating store operated calcium entry. AB - Store-operated calcium entry (SOCE) is a fundamental mechanism of calcium signaling. The mechanisms linking store depletion to SOCE remain controversial, hypothetically involving both diffusible messengers and conformational coupling of stores to channels. Sphingosine 1-phosphate (S1P) is a bioactive sphingolipid that can signal via cell surface G-protein-coupled receptors, but S1P can also act as a second messenger, mobilizing calcium directly via unknown mechanisms. We show here that S1P opens calcium entry channels in human neutrophils (PMNs) and HL60 cells without prior store depletion, independent of G-proteins and of phospholipase C. S1P-mediated entry has the typical divalent cation permeability profile and inhibitor profile of SOCE in PMNs, is fully inhibited by 1 microm Gd3+, and is independent of [Ca2+]i. Depletion of PMN calcium stores by thapsigargin induces S1P synthesis. Inhibition of S1P synthesis by dimethylsphingosine blocks thapsigargin-, ionomycin-, and platelet-activating factor-mediated SOCE despite normal store depletion. We propose that S1P is a "calcium influx factor," linking calcium store depletion to downstream SOCE. PMID- 12746432 TI - Alpha-conotoxins PnIA and [A10L]PnIA stabilize different states of the alpha7 L247T nicotinic acetylcholine receptor. AB - The effects of the native alpha-conotoxin PnIA, its synthetic derivative [A10L]PnIA and alanine scan derivatives of [A10L]PnIA were investigated on chick wild type alpha7 and alpha7-L247T mutant nicotinic acetylcholine receptors (nAChRs) expressed in Xenopus oocytes. PnIA and [A10L]PnIA inhibited acetylcholine (ACh)-activated currents at wtalpha7 receptors with IC50 values of 349 and 168 nm, respectively. Rates of onset of inhibition were similar for PnIA and [A10L]PnIA; however, the rate of recovery was slower for [A10L]PnIA, indicating that the increased potency of [A10L]PnIA at alpha7 receptors is conveyed by its slower rate of dissociation from the receptors. All the alanine mutants of [A10L]PnIA inhibited ACh-activated currents at wtalpha7 receptors. Insertion of an alanine residue between position 5 and 13 and at position 15 significantly reduced the ability of [A10L]PnIA to inhibit ACh-evoked currents. PnIA inhibited the non-desensitizing ACh-activated currents at alpha7-L247T receptors with an IC50 194 nm. In contrast, [A10L]PnIA and the alanine mutants potentiated the ACh-activated current alpha7-L247T receptors and in addition [A10L]PnIA acted as an agonist. PnIA stabilized the receptor in a state that is non-conducting in both the wild type and mutant receptors, whereas [A10L]PnIA stabilized a state that is non-conducting in the wild type receptor and conducting in the alpha7-L247T mutant. These data indicate that the change of a single amino acid side-chain, at position 10, is sufficient to change the toxin specificity for receptor states in the alpha7-L247T mutant. PMID- 12746433 TI - T-cell-derived interleukin-17 regulates the level and stability of cyclooxygenase 2 (COX-2) mRNA through restricted activation of the p38 mitogen-activated protein kinase cascade: role of distal sequences in the 3'-untranslated region of COX-2 mRNA. AB - Although interleukin-17 (IL-17) is the pre-eminent T-cell-derived pro inflammatory cytokine, its cellular mechanism of action remains poorly understood. We explored novel signaling pathways mediating IL-17 induction of the cyclooxygenase-2 (COX-2) gene in human chondrocytes, synovial fibroblasts, and macrophages. In preliminary work, recombinant human (rh) IL-17 stimulated a rapid (5-15 min), substantial (>8-fold), and sustained (>24 h) increase in COX-2 mRNA, protein, and prostaglandin E2 release. Screening experiments with cell-permeable kinase inhibitors (e.g. SB202190 and p38 inhibitor), Western analysis using specific anti-phospho-antibodies to a variety of mitogen-activated protein kinase cascade intermediates, co-transfection studies using chimeric cytomegalovirus driven constructs of GAL4 DNA-binding domains fused to the transactivation domains of transcription factors together with Gal-4 binding element-luciferase reporters, ectopic overexpression of activated protein kinase expression plasmids (e.g. MKK3/6), or transfection experiments with wild-type and mutant COX-2 promoter constructs revealed that rhIL-17 induction of the COX-2 gene was mediated exclusively by the stress-activated protein kinase 2/p38 cascade. A rhIL 17-dependent transcriptional pulse (1.76 +/- 0.11-fold induction) was initiated by ATF-2/CREB-1 transactivation through the ATF/CRE enhancer site in the proximal promoter. However, steady-state levels of rhIL-17-induced COX-2 mRNA declined rapidly (<2 h) to control levels under wash-out conditions. Adding rhIL-17 to transcriptionally arrested cells stabilized COX-2 mRNA for up to 6 h, a process compromised by SB202190. Deletion analysis using transfected chimeric luciferase COX-2 mRNA 3'-untranslated region reporter constructs revealed that rhIL-17 increased reporter gene mRNA stability and protein synthesis via distal regions ( 545 to -1414 bases) of the 3'-untranslated region. This response was mediated entirely by the stress-activated protein kinase 2/p38 cascade. As such, IL-17 can exert direct transcriptional and post-transcriptional control over target proinflammatory cytokines and oncogenes. PMID- 12746434 TI - Inhibition of angiogenesis and angiogenesis-dependent tumor growth by the cryptic kringle fragments of human apolipoprotein(a). AB - Apolipoprotein(a) (apo(a)) contains tandemly repeated kringle domains that are closely related to plasminogen kringle 4, followed by a single kringle 5-like domain and an inactive protease-like domain. Recently, the anti-angiogenic activities of apo(a) have been demonstrated both in vitro and in vivo. However, its effects on tumor angiogenesis and the underlying mechanisms involved have not been fully elucidated. To evaluate the anti-angiogenic and anti-tumor activities of the apo(a) kringle domains and to elucidate their mechanism of action, we expressed the last three kringle domains of apo(a), KIV-9, KIV-10, and KV, in Escherichia coli. The resultant recombinant protein, termed rhLK68, exhibited a dose-dependent inhibition of basic fibroblast growth factor-stimulated human umbilical vein endothelial cell proliferation and migration in vitro and inhibited the neovascularization in chick chorioallantoic membranes in vivo. The ability of rhLK68 to abrogate the activation of extracellular signal-regulated kinases appears to be responsible for rhLK68-mediated anti-angiogenesis. Furthermore, systemic administration of rhLK68 suppressed human lung (A549) and colon (HCT-15) tumor growth in nude mice. Immunohistochemical examination and in situ hybridization analysis of the tumors showed a significant decrease in the number of blood vessels and the reduced expression of vascular endothelial growth factor, basic fibroblast growth factor, and angiogenin, indicating that suppression of angiogenesis may have played a significant role in the inhibition of tumor growth. Collectively, these results suggest that a truncated apo(a), rhLK68, is a potent anti-angiogenic and anti-tumor molecule. PMID- 12746435 TI - Formation of prostaglandins E2 and D2 via the isoprostane pathway: a mechanism for the generation of bioactive prostaglandins independent of cyclooxygenase. AB - It has heretofore been assumed that the cyclooxygenases (COXs) are solely responsible for peostaglandin (PG) synthesis in vivo. An important structural feature of PGH2 formed by COX is the trans-configuration of side chains relative to the prostane ring. Previously, we reported that a series of PG-like compounds termed isoprostanes (IsoPs) are formed in vivo in humans from the free radical catalyzed peroxidation of arachidonate independent of COX. A major difference between these compounds and PGs is that IsoPs are formed from endoperoxide intermediates, the vast majority of which contain side chains that are cis relative to the prostane ring. In addition, unlike the formation of eicosanoids from COX, IsoPs are formed as racemic mixtures because they are generated nonenzymatically. IsoPs containing E- and D-type prostane rings (E2/D2-IsoPs) are one class of IsoPs formed, and we have reported previously that one of the major IsoPs generated is 15-E2t-IsoP (8-iso-PGE2). Unlike PGE2, 15-E2t-IsoP is significantly more unstable in buffered solutions in vitro and undergoes epimerization to PGE2. Analogously, the D-ring IsoP (15-D2c-IsoP) would be predicted to rearrange to PGD2. We now report that compounds identical in all respects to PGE2 and PGD2 and their respective enantiomers are generated in vivo via the IsoP pathway, presumably by epimerization of racemic 15-E2t-IsoP and 15 D2c-IsoP, respectively. Racemic PGE2 and PGD2 were present esterified in phospholipids derived from liver tissue from rats exposed to oxidant stress at levels of 24 +/- 16 and 37 +/- 12 ng/g of tissue, respectively. In addition, racemic PGs, particularly PGD2, were present unesterified in urine from normal animals and humans and represented up to 10% of the total PG detected. Levels of racemic PGD2 increased 35-fold after treatment of rats with carbon tetrachloride to induce oxidant stress. In this setting, PGD2 and its enantiomer generated by the IsoP pathway represented approximately 30% of the total PGD2 present in urine. These findings strongly support the contention that a second pathway exists for the formation of bioactive PGs in vivo that is independent of COX. PMID- 12746436 TI - tbCPSF30 depletion by RNA interference disrupts polycistronic RNA processing in Trypanosoma brucei. AB - Gene expression in eukaryotes requires the post-transcriptional cleavage of mRNA precursors into mature mRNAs. In Trypanosoma brucei, mRNA processing is of particular importance, since most transcripts are derived from polycistronic transcription units. This organization dictates that regulated gene expression is promoter-independent and governed at the posttranscriptional level. We have identified tbCPSF30, a protein containing five CCCH zinc finger motifs, which is a homologue of the cleavage and polyadenylation specificity factor (CPSF) 30-kDa subunit, a component of the machinery required for 3'-end formation in yeast and mammals. Using gene silencing of tbCPSF30 by RNA interference, we demonstrate that this gene is essential in bloodstream and procyclic forms of T. brucei. Interestingly, tbCPSF30-specific RNA interference results in the accumulation of an aberrant tbCPSF30 mRNA species concomitant with depletion of tbCPSF30 protein. tbCPSF30 protein depletion is accompanied by the accumulation of unprocessed tubulin RNAs, implicating tbCPSF30 in polycistronic RNA processing. By genome data base mining, we also identify several other putative components of the T. brucei cleavage and polyadenylation machinery, indicating their conservation throughout eukaryotic evolution. This study is the first to identify and characterize a core component of the T. brucei CPSF and show its involvement in polycistronic RNA processing. PMID- 12746437 TI - Defective discoidin domain structure, subunit assembly, and endoplasmic reticulum processing of retinoschisin are primary mechanisms responsible for X-linked retinoschisis. AB - Retinoschisin is a 24-kDa discoidin domain-containing protein that is secreted from photoreceptor and bipolar cells as a large disulfide-linked multisubunit complex. It functions as a cell adhesion protein to maintain the cellular organization and synaptic structure of the retina. Over 125 different mutations in the RS1 gene are associated with X-linked juvenile retinoschisis, the most common form of early onset macular degeneration in males. To identify molecular determinants important for retinoschisin structure and function and elucidate molecular and cellular mechanisms responsible for X-linked juvenile retinoschisis, we have analyzed the expression, protein folding, disulfide-linked subunit assembly, intracellular localization, and secretion of wild-type retinoschisin, 15 Cys-to-Ser variants and 12 disease-linked mutants. Our studies, together with molecular modeling of the discoidin domain, identify Cys residues involved in intramolecular and intermolecular disulfide bonds essential for protein folding and subunit assembly. We show that misfolding of the discoidin domain, defective disulfide-linked subunit assembly, and inability of retinoschisin to insert into the endoplasmic reticulum membrane as part of the protein secretion process are three primary mechanisms responsible for the loss in the function of retinoschisin as a cell adhesion protein and the pathogenesis of X-linked juvenile retinoschisis. PMID- 12746439 TI - Global iron-dependent gene regulation in Escherichia coli. A new mechanism for iron homeostasis. AB - Organisms generally respond to iron deficiency by increasing their capacity to take up iron and by consuming intracellular iron stores. Escherichia coli, in which iron metabolism is particularly well understood, contains at least 7 iron acquisition systems encoded by 35 iron-repressed genes. This Fe-dependent repression is mediated by a transcriptional repressor, Fur (ferric uptake regulation), which also controls genes involved in other processes such as iron storage, the Tricarboxylic Acid Cycle, pathogenicity, and redox-stress resistance. Our macroarray-based global analysis of iron- and Fur-dependent gene expression in E. coli has revealed several novel Fur-repressed genes likely to specify at least three additional iron-transport pathways. Interestingly, a large group of energy metabolism genes was found to be iron and Fur induced. Many of these genes encode iron-rich respiratory complexes. This iron- and Fur-dependent regulation appears to represent a novel iron-homeostatic mechanism whereby the synthesis of many iron-containing proteins is repressed under iron-restricted conditions. This mechanism thus accounts for the low iron contents of fur mutants and explains how E. coli can modulate its iron requirements. Analysis of 55Fe labeled E. coli proteins revealed a marked decrease in iron-protein composition for the fur mutant, and visible and EPR spectroscopy showed major reductions in cytochrome b and d levels, and in iron-sulfur cluster contents for the chelator treated wild-type and/or fur mutant, correlating well with the array and quantitative RT-PCR data. In combination, the results provide compelling evidence for the regulation of intracellular iron consumption by the Fe2+-Fur complex. PMID- 12746438 TI - Impaired modulation of GABAergic transmission by muscarinic receptors in a mouse transgenic model of Alzheimer's disease. AB - It has long been recognized that muscarinic acetylcholine receptors (mAChRs) are crucial for the control of cognitive processes, and drugs that activate mAChRs are helpful in ameliorating cognitive deficits of Alzheimer's disease (AD). On the other hand, GABAergic transmission in prefrontal cortex (PFC) plays a key role in "working memory" via controlling the timing of neuronal activity during cognitive operations. To test whether the muscarinic and gamma-aminobutyric acid (GABA) system are interconnected in normal cognition and dementia, we examined the muscarinic regulation of GABAergic transmission in PFC of an animal model of AD. Transgenic mice overexpressing a mutant gene for beta-amyloid precursor protein (APP) show behavioral and histopathological abnormalities resembling AD and, therefore, were used as an AD model. Application of the mAChR agonist carbachol significantly increased the spontaneous inhibitory postsynaptic current (sIPSC) frequency and amplitude in PFC pyramidal neurons from wild-type animals. In contrast, carbachol failed to increase the sIPSC amplitude in APP transgenic mice, whereas the carbachol-induced increase of the sIPSC frequency was not significantly changed in these mutants. Similar results were obtained in rat PFC slices pretreated with the beta-amyloid peptide (Abeta). Inhibiting protein kinase C (PKC) blocked the carbachol enhancement of sIPSC amplitudes, implicating the PKC dependence of this mAChR effect. In APP transgenic mice, carbachol failed to activate PKC despite the apparently normal expression of mAChRs. These results show that the muscarinic regulation of GABA transmission is impaired in the AD model, probably due to the Abeta-mediated interference of mAChR activation of PKC. PMID- 12746441 TI - A novel sequence in the coiled-coil domain of Stat3 essential for its nuclear translocation. AB - Stat3 is activated by cytokines and growth factors via specific tyrosine phosphorylation, dimerization, and nuclear translocation. However, the mechanism involved in its nuclear translocation is unclear. In this study, by systematic deletion and site-directed mutagenesis we identified Arg-214/215 in the alpha helix 2 region of the coiled-coil domain of Stat3 as a novel sequence element essential for its nuclear translocation, stimulated by epidermal growth factor as well as by interleukin-6. Furthermore, we identified Arg-414/417 in the DNA binding domain as also required for the nuclear localization of Stat3. This sequence element corresponds to Lys-410/413 of Stat1, a reported sequence for Stat1 nuclear translocation. On the other hand, Leu-411 of Stat3, corresponding to Leu-407 of Stat1, a necessary residue for Stat1 nuclear transport, is not essential for Stat3 nuclear import. The mutant of Arg-214/215 or Arg-414/417 was shown to be tyrosyl-phosphorylated normally but failed to enter the nucleus in response to epidermal growth factor or interleukin-6. The defect, however, can be rescued by the wild-type Stat3 but cannot be compensated by these two mutants. Mutations on Arg-414/417, but not Arg-214/215, destroy the DNA binding activity of Stat3. Our data for the first time identified a sequence element located in the coiled-coil domain that is involved in the ligand-induced nuclear translocation of Stat3. This novel sequence together with a conserved sequence element in the DNA binding domain coordinates to mediate the nuclear translocation of Stat3. PMID- 12746440 TI - Functional distinctions between IMP dehydrogenase genes in providing mycophenolate resistance and guanine prototrophy to yeast. AB - IMP dehydrogenase (IMPDH) catalyzes the rate-limiting step in the de novo synthesis of GTP. Yeast with mutations in the transcription elongation machinery are sensitive to inhibitors of this enzyme such as 6-azauracil and mycophenolic acid, at least partly because of their inability to transcriptionally induce IMPDH. To understand the molecular basis of this drug-sensitive phenotype, we have dissected the expression and function of a four-gene family in yeast called IMD1 through IMD4. We show here that these family members are distinct, despite a high degree of amino acid identity between the proteins they encode. Extrachromosomal copies of IMD1, IMD3, or IMD4 could not rescue the drug sensitive phenotype of IMD2 deletants. When overexpressed, IMD3 or IMD4 weakly compensated for deletion of IMD2. IMD1 is transcriptionally silent and bears critical amino acid substitutions compared with IMD2 that destroy its function, offering strong evidence that it is a pseudogene. The simultaneous deletion of all four IMD genes was lethal unless growth media were supplemented with guanine. This suggests that there are no other essential functions of the IMPDH homologs aside from IMP dehydrogenase activity. Although neither IMD3 nor IMD4 could confer drug resistance to cells lacking IMD2, either alone was sufficient to confer guanine prototrophy. The special function of IMD2 was provided by its ability to be transcriptionally induced and the probable intrinsic drug resistance of its enzymatic activity. PMID- 12746442 TI - Structural (betaalpha)8 TIM barrel model of 3-hydroxy-3-methylglutaryl-coenzyme A lyase. AB - This study describes three novel homozygous missense mutations (S75R, S201Y, and D204N) in the 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) lyase gene, which caused 3 hydroxy-3-methylglutaric aciduria in patients from Germany, England, and Argentina. Expression studies in Escherichia coli show that S75R and S201Y substitutions completely abolished the HMG-CoA lyase activity, whereas D204N reduced catalytic efficiency to 6.6% of the wild type. We also propose a three dimensional model for human HMG-CoA lyase containing a (betaalpha)8 (TIM) barrel structure. The model is supported by the similarity with analogous TIM barrel structures of functionally related proteins, by the localization of catalytic amino acids at the active site, and by the coincidence between the shape of the substrate (HMG-CoA) and the predicted inner cavity. The three novel mutations explain the lack of HMG-CoA lyase activity on the basis of the proposed structure: in S75R and S201Y because the new amino acid residues occlude the substrate cavity, and in D204N because the mutation alters the electrochemical environment of the active site. We also report the localization of all missense mutations reported to date and show that these mutations are located in the beta sheets around the substrate cavity. PMID- 12746443 TI - The chloride channel ClC-4 contributes to endosomal acidification and trafficking. AB - Mutations in the gene coding for the chloride channel ClC-5 cause Dent's disease, a disease associated with proteinuria and renal stones. Studies in ClC-5 knockout mice suggest that this phenotype is related to defective endocytosis of low molecular weight proteins and membrane proteins by the renal proximal tubule. In this study, confocal micrographs of proximal tubules and cultured epithelial cells revealed that the related protein ClC-4 is expressed in endosomal membranes suggesting that this channel may also contribute to the function of this organelle. In support of this hypothesis, specific disruption of endogenous ClC-4 expression by transfection of ClC-4 antisense cDNA acidified endosomal pH and altered transferrin trafficking in cultured epithelial cells to the same extent as the specific disruption of ClC-5. Both channels can be co-immunoprecipitated, arguing that they may partially contribute to endosomal function as a channel complex. These studies prompt future investigation of the role of ClC-4 in renal function in health and in Dent's disease. Future studies will assess whether the severity of Dent's disease relates not only to the impact of particular mutations on ClC-5 but also on the consequences of those mutations on the functional expression of ClC-4. PMID- 12746444 TI - The ATP hydrolysis cycle of the nucleotide-binding domain of the mitochondrial ATP-binding cassette transporter Mdl1p. AB - The ABC transporter Mdl1p, a structural and functional homologue of the transporter associated with antigen processing (TAP) plays an important role in intracellular peptide transport from the mitochondrial matrix of Saccharomyces cerevisiae. To characterize the ATP hydrolysis cycle of Mdl1p, the nucleotide binding domain (NBD) was overexpressed in Escherichia coli and purified to homogeneity. The isolated NBD was active in ATP binding and hydrolysis with a turnover of 25 ATP per minute and a Km of 0.6 mm and did not show cooperativity in ATPase activity. However, the ATPase activity was non-linearly dependent on protein concentration (Hill coefficient of 1.7), indicating that the functional state is a dimer. Dimeric catalytic transition states could be trapped either by incubation with orthovanadate or beryllium fluoride, or by mutagenesis of the NBD. The nucleotide composition of trapped intermediate states was determined using [alpha-32P]ATP and [gamma-32P]ATP. Three different dimeric intermediate states were isolated, containing either two ATPs, one ATP and one ADP, or two ADPs. Based on these experiments, it was shown that: (i) ATP binding to two NBDs induces dimerization, (ii) in all isolated dimeric states, two nucleotides are present, (iii) phosphate can dissociate from the dimer, (iv) both nucleotides are hydrolyzed, and (v) hydrolysis occurs in a sequential mode. Based on these data, we propose a processive-clamp model for the catalytic cycle in which association and dissociation of the NBDs depends on the status of bound nucleotides. PMID- 12746445 TI - Hepatocyte growth factor preserves beta cell mass and mitigates hyperglycemia in streptozotocin-induced diabetic mice. AB - Type I diabetes is an autoimmune disease that results in destructive depletion of the insulin-producing beta cells in the islets of Langerhans in pancreas. With the knowledge that hepatocyte growth factor (HGF) is a potent survival factor for a wide variety of cells, we hypothesized that supplementation of HGF may provide a novel strategy for protecting pancreatic beta cells from destructive death and for preserving insulin production. In this study, we demonstrate that expression of the exogenous HGF gene preserved insulin excretion and mitigated hyperglycemia of diabetic mice induced by streptozotocin. Blood glucose levels were significantly reduced in mice receiving a single intravenous injection of naked HGF gene at various time points after streptozotocin administration. Consistently, HGF concomitantly increased serum insulin levels in diabetic mice. Immunohistochemical staining revealed a marked preservation of insulin-producing beta cells by HGF in the pancreatic islets of the diabetic mice. This beneficial effect of HGF was apparently mediated by both protection of beta cells from death and promotion of their proliferation. Delivery of HGF gene in vivo induced pro survival Akt kinase activation and Bcl-xL expression in the pancreatic islets of diabetic mice. These findings suggest that supplementation of HGF to prevent beta cells from destructive depletion and to promote their proliferation might be an effective strategy for ameliorating type I diabetes. PMID- 12746446 TI - Glucose stimulates the tyrosine phosphorylation of Crk-associated substrate in pancreatic beta-cells. AB - Several years ago, we demonstrated that glucose induced tyrosine phosphorylation of a 125-kDa protein (p125) in pancreatic beta-cells (Konrad, R. J., Dean, R. M., Young, R. A., Bilings, P. C., and Wolf, B. A. (1996) J. Biol. Chem. 271, 24179 24186). Glucose induced p125 tyrosine phosphorylation in beta-TC3 insulinoma cells, beta-HC9 cells, and in freshly isolated rat islets, whereas increased tyrosine phosphorylation was not observed with other fuel secretagogues. Initial efforts to identify p125 were unsuccessful, so a new approach was taken. The protein was purified from betaTC6,F7 cells via an immunodepletion method. After electrophoresis and colloidal Coomassie Blue staining, the area of the gel corresponding to p125 was excised and subjected to tryptic digestion. Afterward, mass spectrometry was performed and the presence of Crk-associated substrate (Cas) was detected. Commercially available antibodies against Cas were obtained and tested directly in beta-cells, confirming glucose-induced tyrosine phosphorylation of Cas. Further experiments demonstrated that in beta-cells the glucose-induced increase in Cas tyrosine phosphorylation occurs immediately and is not accompanied by increased focal adhesion kinase tyrosine phosphorylation. Finally, it is also demonstrated via Western blotting that Cas is present in normal isolated rat islets. Together, these results show that the identity of the previously described p125 beta-cell protein is Cas and that Cas undergoes rapid glucose-induced tyrosine phosphorylation in beta-cells. PMID- 12746447 TI - Crystal structure of the tRNA processing enzyme RNase PH from Aquifex aeolicus. AB - RNase PH is one of the exoribonucleases that catalyze the 3' end processing of tRNA in bacteria. RNase PH removes nucleotides following the CCA sequence of tRNA precursors by phosphorolysis and generates mature tRNAs with amino acid acceptor activity. In this study, we determined the crystal structure of Aquifex aeolicus RNase PH bound with a phosphate, a co-substrate, in the active site at 2.3-A resolution. RNase PH has the typical alpha/beta fold, which forms a hexameric ring structure as a trimer of dimers. This ring structure resembles that of the polynucleotide phosphorylase core domain homotrimer, another phosphorolytic exoribonuclease. Four amino acid residues, Arg-86, Gly-124, Thr-125, and Arg-126, of RNase PH are involved in the phosphate-binding site. Mutational analyses of these residues showed their importance in the phosphorolysis reaction. A docking model with the tRNA acceptor stem suggests how RNase PH accommodates substrate RNAs. PMID- 12746448 TI - Normal sorting but defective endocytosis of the low density lipoprotein receptor in mice with autosomal recessive hypercholesterolemia. AB - Autosomal recessive hypercholesterolemia (ARH) is a genetic form of hypercholesterolemia that clinically resembles familial hypercholesterolemia (FH). As in FH, the rate of clearance of circulating low density lipoprotein (LDL) by the LDL receptor (LDLR) in the liver is markedly reduced in ARH. Unlike FH, LDL uptake in cultured fibroblasts from ARH patients is normal or only slightly impaired. The gene defective in ARH encodes a putative adaptor protein that has been implicated in linking the LDLR to the endocytic machinery. To determine the role of ARH in the liver, ARH-deficient mice were developed. Plasma levels of LDL-cholesterol were elevated in the chow-fed Arh-/- mice (83 +/- 8 mg/dl versus 68 +/- 8 mg/dl) but were lower than those of mice expressing no LDLR (Ldlr-/-) (197 +/- 8 mg/dl). Cholesterol feeding elevated plasma cholesterol levels in both strains. The fractional clearance rate of radiolabeled LDL was reduced to similar levels in the Arh-/- and Ldlr-/- mice, whereas the rate of removal of alpha2-macroglobulin by the LDLR-related protein, which also interacts with ARH, was unchanged. Immunolocalization studies revealed that a much greater proportion of immunodetectable LDLR, but not LDLR-related protein, was present on the sinusoidal surface of hepatocytes in the Arh-/- mice. Taken together, these results are consistent with ARH playing a critical and specific role in LDLR endocytosis in the liver. PMID- 12746450 TI - Mutations in the met oncogene unveil a "dual switch" mechanism controlling tyrosine kinase activity. AB - The met oncogene, encoding the high affinity hepatocyte growth factor receptor, is the only known gene inherited in human cancer that is invariably associated with somatic duplication of the mutant locus. Intriguingly, mutated Met requires ligand stimulation in order to unleash its transforming potential. Furthermore, individuals bearing a germ line met mutation develop cancer only late in life and with incomplete penetrance. To date, there is no molecular explanation for this unique behavior, which is unusual for a dominant oncogene. Here we investigate the molecular mechanisms underlying met oncogenic conversion by generating antibodies specific for the differently phosphorylated forms of the Met protein. Using these antibodies, we show that activation of wild-type Met is achieved through sequential phosphorylation of Tyr1235 and Tyr1234 in the activation loop and that mutagenesis of either tyrosine dramatically impairs kinase function. Surprisingly, oncogenic Met mutants never become phosphorylated on Tyr1234 despite their high enzymatic activity, and mutagenesis of Tyr1234 does not affect their biochemical or biological function. By analyzing the enzymatic properties of the mutant proteins in different conditions, we demonstrate that oncogenic mutations do not elicit constitutive kinase activation but simply overcome the requirement for the second phosphorylation step, thus reducing the threshold for activation. In the presence of activating signals, these mutations result therefore in a dynamic imbalance toward the active conformation of the kinase. This explains why mutant met provides an oncogenic predisposition but needs a second activating "hit," provided by sustained ligand stimulation or receptor overexpression, to achieve a fully transformed phenotype. PMID- 12746449 TI - A carboxyl-terminal mutation of the epidermal growth factor receptor alters tyrosine kinase activity and substrate specificity as measured by a fluorescence polarization assay. AB - The expression of certain COOH-terminal truncation mutants of the epidermal growth factor receptor (EGFR) can lead to cell transformation, and with ligand stimulation, a broader spectrum of phosphorylated proteins appears compared with EGF-treated cells expressing wild-type EGFR. Accordingly, it has been proposed that elements within the COOH terminus may determine substrate specificity of the EGFR tyrosine kinase (Decker, S. J., Alexander, C., and Habib, T. (1992) J. Biol. Chem. 267, 1104-1108; Walton, G. M., Chen, W. S., Rosenfeld, M. G., and Gill, G. N. (1990) J. Biol. Chem. 265, 1750-1754). To address this hypothesis, we analyzed in vitro the steady-state kinetic parameters for phosphorylation of several substrates by both wild-type EGFR and an oncogenic EGFR mutant (the ct1022 mutant) truncated at residue 1022. The substrates included: (i) a phospholipase C gamma fragment (residues 530-850); (ii) the 46-kDa isoform of the Shc adapter protein; (iii) a 13-residue peptide mimic for the region around the major autophosphorylation tyrosine and the Shc binding site (the Y1173 peptide); (iv) a poly(Glu,Tyr) 4:1 copolymer; and (v) the 8-residue peptide, angiotensin II. Our data demonstrate that the steady-state kinetic parameters for the ct1022 mutant differ from those of the wild-type enzyme, and the differences are substrate dependent. These results support the concept that this oncogenic truncation/mutation alters EGFR substrate specificity, rather than causing a general alteration of activity. We performed the experiments using a non radioactive fluorescence polarization assay that quantifies the degree of phosphorylation of peptide as well as natural substrates. The results are consistent with those from the traditional [gamma-32P]ATP/filtration assay. PMID- 12746451 TI - Pulmonary inflammation and edema induced by phospholipase A2: global gene analysis and effects on aquaporins and Na+/K+-ATPase. AB - Victims of snakebite quickly succumb to severe respiratory failure, which can be fatal if left untreated. One of the most toxic components of snake venom is phospholipase A2 (PLA2; EC 3.1.1.4). PLA2 isolated from the elapid, Naja sputatrix, induced pulmonary inflammation and edema when administered intravenously and intratracheally to rats. Analysis of pulmonary gene expression profiles using oligonucleotide microarrays revealed 60 genes whose expression was altered by at least 3-fold in response to intratracheal instillation of PLA2 for 3 h as compared with controls. In addition to genes encoding cytokines and chemokines responsible for inflammatory processes, the Na+/K+-ATPase gene has been found to be involved in edema formation. Real-time PCR, Western blot, and immunohistochemical analyses confirmed that the expression of AQP1 and AQP5 mRNAs and proteins was decreased. Besides providing an experimental model for studies on the pathophysiology of the lung, this investigation yields a clue to the mechanisms by which endogenous PLA2s could mediate inflammation in conditions such as allergy and rheumatoid arthritis. PMID- 12746452 TI - cFLIP-L inhibits p38 MAPK activation: an additional anti-apoptotic mechanism in bile acid-mediated apoptosis. AB - In cholestasis, toxic bile acids accumulate within the liver inducing hepatocyte apoptosis, which exacerbates liver injury. Although bile acids activate both death receptors and mitogen-activated kinase (MAPK) pathways, the mechanistic link between death receptor signaling and MAPK activation in bile acid apoptosis remains unclear. The aim of this study was to ascertain if MAPKs contribute to bile acid cytotoxicity. Although deoxycholate induced apoptosis and activated all three classic mediators of the MAPK pathways including JNK 1/2, p38, and p42/44, only p38 MAPK inhibition attenuated apoptosis. Suppressing FADD expression with siRNA or employing a caspase inhibitor, zVAD-fmk, did not block p38 MAPK activation suggesting its activation was not death receptor-dependent. Unexpectedly, expression of cFLIP-L in a stably transfected cell line blocked apoptosis and p38 MAPK phosphorylation. Based on these data we postulated a direct effect of cFLIP on p38 MAPK activation. The nonphosphorylated but not the phosphorylated/active form of p38 MAPK co-immunoprecipitated with cFLIP-L. In reverse immunoprecipitation experiments, cFLIP-L long but not cFLIP-S co immunoprecipitate with p38 MAPK. In conclusion, these data suggest that cFLIP-L exerts its anti-apoptotic activity, in part, by inhibiting p38 MAPK activation, an additional anti-apoptotic effect for this protein. PMID- 12746453 TI - Cyclin D1 splice variants. Differential effects on localization, RB phosphorylation, and cellular transformation. AB - Cyclin D1 is a proto-oncogene that functions by inactivation of the retinoblastoma tumor suppressor protein, RB. A common polymorphism in the cyclin D1 gene is associated with the production of an alternate transcript of cyclin D1, termed cyclin D1b. Both the polymorphism and the variant transcript are associated with increased risk for multiple cancers and the severity of a given cancer; however, the underlying activities of cyclin D1b have not been elucidated relative to the canonical cyclin D1a. Because cyclin D1b does not possess the threonine 286 phosphorylation site required for nuclear export and regulated degradation, it has been hypothesized to encode a stable nuclear protein that would constitutively inactivate the RB pathway. Surprisingly, we find that cyclin D1b protein does not inappropriately accumulate in cells and exhibits stability comparable to cyclin D1a. As expected, the cyclin D1b protein was constitutively localized in the nucleus, whereas cyclin D1a was exported to the cytoplasm in S phase. Despite enhanced nuclear localization, we find that cyclin D1b is a poor catalyst of RB phosphorylation/inactivation. However, cyclin D1b potently induced cellular transformation in contrast to cyclin D1a. In summary, we demonstrate that cyclin D1b specifically disrupts contact inhibition in a manner distinct from cyclin D1a. These data reveal novel roles for d-type cyclins in tumorigenesis. PMID- 12746454 TI - Detection of tRNA-like structure through RNase P cleavage of viral internal ribosome entry site RNAs near the AUG start triplet. AB - The 9600-base RNA genome of hepatitis C virus (HCV) has an internal ribosome entry site (IRES) in its first 370 bases, including the AUG start triplet at bases 342-344. Structural elements of this and other IRES domains substitute for a 5' terminal cap structure in protein synthesis. Recent work (Nadal, A., Martell, M., Lytle, J. R., Lyons, A. J., Robertson, H. D., Cabot, B., Esteban, J. I., Esteban, R., Guardia, J., and Gomez, J. (2002) J. Biol. Chem. 277, 30606 30613) has demonstrated that the host pre-tRNA processing enzyme, RNase P, can cleave the HCV RNA genome at a site in the IRES near the AUG initiator triplet. Although this step is unlikely to be part of the HCV life cycle, such a reaction could indicate the presence of a tRNA-like structure in this IRES. Because susceptibility to cleavage by mammalian RNase P is a strong indicator of tRNA like structure, we have conducted the studies reported here to test whether such tRNA mimicry is unique to HCV or is a general property of IRES structure. We have assayed IRES domains of several viral RNA genomes: two pestiviruses related to HCV, classical swine fever virus and bovine viral diarrhea virus; and two unrelated viruses, encephalomyocarditis virus and cricket paralysis virus. We have found similarly placed RNase P cleavage sites in these IRESs. Thus a tRNA like domain could be a general structural feature of IRESs, the first IRES structure to be identified with a functional correlate. Such tRNA-like features could be recognized by pre-existing ribosomal tRNA-binding sites as part of the IRES initiation cycle. PMID- 12746455 TI - Erythropoietin receptors associate with a ubiquitin ligase, p33RUL, and require its activity for erythropoietin-induced proliferation. AB - The proliferation and survival of hematopoietic cells is strictly regulated by cytokine growth factors that act through receptors of the Type I cytokine receptor family, including erythropoietin (Epo) and its receptor, EpoR. Mitogenic signaling by these receptors depends on activation of Jak tyrosine kinases. However, other required components of this pathway have not been fully identified. In a screen for proteins that interact with EpoR and Jak2, we identified a novel member of the U-box family of ubiquitin ligases. This receptor associated ubiquitin ligase, RUL, co-precipitated with EpoR from mammalian cells and mediated ubiquitination of EpoR. Also, endogenously expressed RUL was rapidly and transiently phosphorylated on serine after cytokine treatment of factor dependent hematopoietic cells. Expression of ubiquitin ligase-deficient mutants of RUL inhibited Epo-induced expression of c-myc and bcl-2, two immediate-early genes normally associated with Epo-induced cell growth. Consistent with that finding, expression of mutant RUL also inhibited Epo-dependent proliferation and survival of factor-dependent cells. Together, these observations suggest that RUL is a required component of mitogenic signaling by EpoR. We also show that RUL is phosphorylated in response to growth factors that act through non-cytokine receptors, suggesting that RUL may function as a common regulator of mitogenesis. PMID- 12746456 TI - Oligomerization of dopamine transporters visualized in living cells by fluorescence resonance energy transfer microscopy. AB - To examine the oligomeric state and trafficking of the dopamine transporter (DAT) in different compartments of living cells, human DAT was fused to yellow (YFP) or cyan fluorescent protein (CFP). YFP-DAT and CFP-DAT were transiently and stably expressed in porcine aortic endothelial (PAE) cells, human embryonic kidney (HEK) 293 cells, and an immortalized dopaminergic cell line 1RB3AN27. Fluorescence microscopic imaging of cells co-expressing YFP-DAT and CFP-DAT revealed fluorescence resonance energy transfer (FRET) between CFP and YFP, which is consistent with an intermolecular interaction of DAT fusion proteins. FRET signals were detected between CFP- and YFP-DAT located at the plasma membrane and in intracellular membrane compartments. Phorbol esters or amphetamine induced the endocytosis of YFP/CFP-DAT to early and recycling endosomes, identified by Rab5, Rab11, Hrs and EEA.1 proteins. Interestingly, however, DAT was mainly excluded from Rab5- and Hrs-containing microdomains within the endosomes. The strongest FRET signals were measured in endosomes, indicative of efficient oligomerization of internalized DAT. The intermolecular DAT interactions were confirmed by co immunoprecipitation. A DAT mutant that was retained in the endoplasmic reticulum (ER) after biosynthesis was used to show that DAT is oligomeric in the ER. Moreover, co-expression of an ER-retained DAT mutant and wild-type DAT resulted in the retention of wild-type DAT in the ER. These data suggest that DAT oligomers are formed in the ER and then are constitutively maintained both at the cell surface and during trafficking between the plasma membrane and endosomes. PMID- 12746457 TI - Functional analysis of the rat N-methyl-D-aspartate receptor 2A promoter: multiple transcription starts points, positive regulation by Sp factors, and translational regulation. AB - N-Methyl-d-aspartate (NMDA) receptor subunit 2A (NR2A) is an important modulatory component of the NMDA subtype of glutamate receptors. To investigate the transcription mechanism of the NR2A gene, we cloned the 5'-flanking sequence from a rat genomic library. RNA mapping with rat brain RNA revealed two sets of major and several minor transcription start points in a single exon of 1140 bp. Reporter gene and mutation studies indicated that core promoter activity resided in exon 1, whereas the 5'-flanking sequence up to 1.5 kb showed no significant impact on promoter activity. Fragments containing minor transcription start points were able to drive a reporter gene in transfected cells and produce nascent RNAs in an in vitro transcription system. All fragments tested showed more promoter activity in dissociated neurons of the rat embryonic cerebrocortex and cell lines expressing NR2A mRNA than that in glial cultures and non-neuronal cells. Within exon 1 there are three GC-box elements that displayed distinct binding affinity to both Sp1- and Sp4-like factors. Overexpression of Sp1 or Sp4, but not Sp3, significantly increased the activity of the promoter containing these elements. Inclusion of exon 2 and 3 sequences, which contain five short open-reading frames, attenuated promoter-driven reporter activity more than 3 fold but attenuated the level of reporter mRNA less than 1.4-fold. Our results suggest that the core promoter of the rat NR2A gene requires exon 1, that Sp factors positively regulate this core promoter, and that a post-transcriptional mechanism may negatively regulate expression of the gene. PMID- 12746459 TI - Vpu exerts a positive effect on HIV-1 infectivity by down-modulating CD4 receptor molecules at the surface of HIV-1-producing cells. AB - Human immunodeficiencey virus, type 1 (HIV-1) encodes three proteins, Nef, Vpu, and gp160, that down-modulate surface expression of the CD4 receptor during viral infection. In the present study, we have investigated the role of CD4 down modulation in the HIV-1 infection cycle, primarily from the perspective of Vpu function. We report here that, like Nef, Vpu-mediated CD4 degradation modulates positively HIV-1 infectivity. Our data reveal that accumulation of CD4 at the cell surface of Vpu-deficient HIV-1-producing cells leads to an efficient recruitment of CD4 into virions and to an impairment of viral infectivity. This CD4-mediated inhibition of viral infectivity was not observed when a CD4 mutant unable to bind Env gp120 was used or when VSV-G glycoprotein was utilized to pseudotype viruses, suggesting that an interaction between CD4 and gp120 is required for interference. Indeed, protein analysis of Vpu-defective viral particles reveals that CD4 recruitment is associated with an increased formation of gp120-CD4 complexes at the virion surface. Interestingly, we did not detect any difference at the level of total virion-associated Env glycoproteins between wild-type and Vpu-defective virus, indicating that accumulation of CD4 at the cell surface and recruitment of CD4 into Vpu-defective HIV-1 particles exert a negative effect on viral infectivity, most likely by promoting the formation of nonfunctional gp120-CD4 complexes at the virion surface. Finally, we show that both Vpu- and Nef-induced CD4 down-modulation activities are required for production of fully infectious particles in CD4+ T cell lines and primary cells, an observation that has clear implications for viral spread in vivo. PMID- 12746458 TI - Calcium-regulated interaction of Sgt1 with S100A6 (calcyclin) and other S100 proteins. AB - S100A6 (calcyclin), a small calcium-binding protein from the S100 family, interacts with several target proteins in a calcium-regulated manner. One target is Calcyclin-Binding Protein/Siah-1-Interacting Protein (CacyBP/SIP), a component of a novel pathway of beta-catenin ubiquitination. A recently discovered yeast homolog of CacyBP/SIP, Sgt1, associates with Skp1 and regulates its function in the Skp1/Cullin1/F-box complex ubiquitin ligase and in kinetochore complexes. S100A6-binding domain of CacyBP/SIP is in its C-terminal region, where the homology between CacyBP/SIP and Sgt1 is the greatest. Therefore, we hypothesized that Sgt1, through its C-terminal region, interacts with S100A6. We tested this hypothesis by performing affinity chromatography and chemical cross-linking experiments. Our results showed that Sgt1 binds to S100A6 in a calcium-regulated manner and that the S100A6-binding domain in Sgt1 is comprised of 71 C-terminal residues. Moreover, S100A6 does not influence Skp1-Sgt1 binding, a result suggesting that separate Sgt1 domains are responsible for interactions with S100A6 and Skp1. Sgt1 binds not only to S100A6 but also to S100B and S100P, other members of the S100 family. The interaction between S100A6 and Sgt1 is likely to be physiologically relevant because both proteins were co-immunoprecipitated from HEp-2 cell line extract using monoclonal anti-S100A6 antibody. Phosphorylation of the S100A6-binding domain of Sgt1 by casein kinase II was inhibited by S100A6, a result suggesting that the role of S100A6 binding is to regulate the phosphorylation of Sgt1. These findings suggest that protein ubiquitination via Sgt1-dependent pathway can be regulated by S100 proteins. PMID- 12746460 TI - Sphingolipid requirement for generation of a functional v1 component of the vacuolar ATPase. AB - There has been no previous indication that vacuolar ATPases (V-ATPases) require sphingolipids for function. Here we show, by using Saccharomyces cerevisiae sur4Delta and fen1Delta cells, that sphingolipids with a C26 acyl group are required for generating V1 domains with ATPase activity. Sphingolipids in sur4Delta cells contain C22 and C24 acyl groups instead of C26 acyl groups whereas about 30% of the sphingolipids in fen1Delta cells have C26 acyl groups and the rest have C22 and C24 acyl groups. sur4Delta cells have several phenotypes (vacuolar membrane ATPase, Vma-) that indicate a defect in the V ATPase, and vacuoles purified from sur4Delta cells have little to no ATPase activity. These phenotypes are less pronounced in fen1Delta cells, consistent with the idea that the C26 acyl group in sphingolipids is necessary for V-ATPase activity. Other results show that the two V-ATPase domains, V1 and V0, are assembled and delivered to the vacuolar membrane in sur4Delta cells similar to wild-type cells. In vitro assembly studies show that V1 from sur4Delta cells associates with wild-type V0 but the complex lacks V-ATPase activity, indicating that V1 is defective. Reciprocal experiments with V0 from sur4Delta cells show that it is normal. We conclude that sphingolipids with a C26 acyl group are required for generating fully functional V1 domains. PMID- 12746462 TI - Meeting report: NHLBI symposium on phenotyping: mouse cardiovascular function and development. PMID- 12746463 TI - Three-dimensional reconstruction of gene expression patterns during cardiac development. AB - The study of the genetic regulation of embryonic development requires the three dimensional (3D) mapping of gene expression at the microscopic level. Despite the recent burst in the number of methods focusing on 3D reconstruction of embryonic specimens, an adequate and accessible 3D reconstruction protocol for the visualization of patterns of gene expression is lacking. In this communication we describe a protocol that was developed for the 3D visualization of patterns of gene expression determined by in situ hybridization (ISH) on serial sections. The method still requires tissue sectioning, due to penetration limits of the specific staining agents into whole embryo preparations. With regard to expenditure of resources, i.e., hardware, software, and time, the protocol is relatively undemanding. Because the variation between specimens requires the visualization of multiple specimens per stage, it was decided to "do more, less well." The current protocol, therefore, results in reconstructions of sufficient, but not the highest, quality. The use of the protocol is demonstrated on a series of serially sectioned mouse hearts, ranging from embryonic day 8.5 to 14.5. The myocardium of the hearts was identified by ISH using a mixture of specific mRNA probes and reconstructed. PMID- 12746464 TI - Evaluation and applications of radiotelemetry in small laboratory animals. AB - Radiotelemetry is the "state of the art" for monitoring physiological functions in awake and freely moving laboratory animals, while minimizing stress artifacts. For researchers, especially those in the fields of pharmacology and toxicology, the technique provides a valuable tool for defining the physiological and pathophysiological consequences derived from advances molecular, cellular, and tissue biology and in predicting the effectiveness and safety of new compounds in humans. There is ample evidence that radiotelemetry systems for measuring physiological functions has been sufficiently validated. Today, the technology is an important tool for collection of a growing number of physiological parameters, for contributing to animal welfare (reduction and refinement alternatives), and for reducing overall animal research costs. PMID- 12746465 TI - Electrophysiological phenotyping in genetically engineered mice. AB - Advances in transgene and gene targeting technology have enabled sophisticated manipulation of the mouse genome, providing important insights into the molecular mechanisms underlying cardiac conduction, arrhythmogenesis, and sudden cardiac death. The mouse is currently the principal mammalian model for studying biological processes, particularly related to cardiac pathophysiology. Murine models have been engineered harboring gene mutations leading to inherited structural and electrical disorders of the heart due to transcription factor mutations, connexin protein defects, and G protein and ion channelopathies. These mutations lead to phenotypes reminiscent of human clinical disease states including congenital heart defects, cardiomyopathies, and long-QT syndrome, creating models of human electrophysiological disease. Functional analyses of the underlying molecular mechanisms of resultant phenotypes require appropriate and sophisticated experimental methodology. This paper reviews current in vivo murine electrophysiology study techniques and genetic mouse models pertinent to human arrhythmia disorders. PMID- 12746466 TI - Exercise assessment of transgenic models of human cardiovascular disease. AB - Exercise provides one of the most severe, yet physiological, stresses to the intact cardiovascular system and is a major determinant of the utilization of metabolic substrates. The adaptations to exercise are the result of a coordinated response of multiple organ systems, including cardiovascular, pulmonary, endocrine-metabolic, immunologic, and skeletal muscle. With the proliferation of genetically altered murine models of cardiovascular disease, the importance of developing methods of accurate physiological phenotyping is critical. There are numerous examples of transgenic models in which the baseline cardiovascular phenotype is unchanged or minimally changed from the wild type, only to become manifest during the stress of exercise testing. In this review, we cover the basics of the murine cardiovascular response to exercise and the importance of attending to strain differences, compare different exercise methodologies (constant workload treadmill, incremental workload treadmill, swimming) and hemodynamic monitoring systems, and examine the murine response to exercise conditioning. Several examples where exercise studies have contributed to the elucidation of cardiovascular phenotypes are reviewed: the beta-adrenergic receptor knockouts, phospholamban knockout, dystrophin knockout (mdx), and the mutant alpha-myosin heavy chain (R403Q) transgenic. PMID- 12746467 TI - Use of echocardiography for the phenotypic assessment of genetically altered mice. AB - Transgenic mice displaying abnormalities in cardiac development and function represent a powerful new tool for understanding molecular mechanisms underlying normal cardiovascular function and the pathophysiological bases of human cardiovascular disease. Complete cardiac evaluation of phenotypic changes in mice requires the ability to noninvasively assess cardiovascular structure and function in a serial manner. However, the small mouse heart beating at rates in excess of 500 beats/min presents unique methodological challenges. Two dimensional and Doppler echocardiography have been recently used as effective, noninvasive tools for murine imaging, because quality images of cardiac structures and valvular flows can be obtained with newer high-frequency transthoracic transducers. We will discuss the use of echocardiography for the assessment of 1) left ventricular (LV) chamber dimensions and wall thicknesses, 2) LV mass, 3) improved endocardial border delineation using contrast echocardiography, 4) LV contractility using ejection phase indices and load independent indices, 5) vascular properties, and 6) LV diastolic performance. Evaluation of cardiovascular performance in closed chest mice is feasible in a variety of murine models using Doppler echocardiographic imaging. PMID- 12746470 TI - Crescentic glomerulonephritis and subepidermal blisters with autoantibodies to alpha5 and alpha6 chains of type IV collagen. AB - We describe a novel autoimmune disease characterized by severe subepidermal bullous eruption and crescentic glomerulonephritis with autoantibodies directed against the noncollagenous domain of the alpha5 and alpha6 chains of type IV collagen. Biopsy of perilesional skin revealed a subepidermal blister with marked polymorphonuclear infiltrate with linear deposits of IgA and C3. Light microscopy of a kidney biopsy specimen revealed a crescentic glomerulonephritis, and immunofluorescence microscopy showed linear basement membrane staining for IgA (3+), C3 (1+), and IgG (1+). No electron-dense deposits were observed by transmission electron microscopy. The patient's autoantibodies reacted with normal human skin and kidney: IgA (3+) and IgG (1+) antibodies stained the basement membrane zones of skin, renal glomerulus, and some tubules. The identity of the target antigen was determined by immunochemical analyses of candidate antigens using the patient's autoantibodies. The patient's IgA and IgG autoantibodies reacted with a 185- to 190-kDa antigen from a human dermal extract that was distinguished from the other dermal or epidermal antigens, including the 145- to 290-kDa (type VII collagen) epidermolysis bullosa acquisita antigen, the 165- to 200-kDa alpha3 laminin mucous membrane cicatricial pemphigoid antigen, and the 230-kDa and the 180-kDa bullous pemphigoid antigens. Patient's IgA and IgG autoantibodies further reacted with the alpha5(IV) and weakly with the alpha6(IV) chains of type IV collagen by Western blot and ELISA. This report expands the repertoire of bullous skin disorders and provides an explanation for the association of anti-type IV collagen autoantibodies and glomerulonephritis with subepidermal blisters. PMID- 12746468 TI - Dietary fat interacts with QTLs controlling induction of Pgc-1 alpha and Ucp1 during conversion of white to brown fat. AB - To identify novel regulatory factors controlling induction of the brown adipocyte specific mitochondrial uncoupling protein (Ucp1) mRNA in the retroperitoneal white fat depot, we previously mapped quantitative trait loci (QTLs) that control this trait to chromosomes 2, 3, 8, and 19. Since the peroxisome proliferator activator receptor-gamma coactivator-1alpha (PGC-1alpha) regulates Ucp1 and other genes of energy metabolism, we have evaluated whether the QTLs controlling Ucp1 mRNA levels also modulate Pgc-1alpha mRNA levels by analysis of backcross progeny from the A/J and C57BL/6J strains of mice. The results indicate that a locus on chromosome 3 orchestrates expression of Pgc-1alpha and Ucp1 in retroperitoneal fat of mice fed a low-fat diet; however, the effect of this locus on Pgc-1alpha is lost, and a significant correlation between Ucp1 and Pgc-1alpha is severely reduced in mice fed a high-fat diet. An additional QTL located on chromosome 5 has also been identified for the selective regulation of Ucp1 mRNA levels. Similar to the effects of a high-fat diet on the chromosome 3 QTL, linkage of the chromosome 5 QTL is also lost in mice on a high-fat diet. Thus dietary fat has a profound influence on PGC-1alpha-regulated pathways controlling energy metabolism in white fat. The allelic variation observed in the regulation of Ucp1 and Pgc 1alpha expression in brown adipocytes of white fat but not interscapular brown fat suggests that fundamentally different regulatory mechanisms exist to control the thermogenic capacities of these tissues. PMID- 12746471 TI - Heparanase expression correlates with invasion and poor prognosis in gastric cancers. AB - Degradation of basement membrane and extracellular matrix structures are important features of the metastatic process of malignant tumors. Human heparanase degrades heparan sulfate proteoglycans, which represent the main components of basement membranes and the extracellular matrix. Because of the role of heparanase in tumor invasion and metastasis, we examined heparanase expression in primary gastric cancers and in cell lines derived from gastric cancers by immunohistochemistry and RT-PCR, respectively. Four of seven gastric cancer cell lines showed heparanase mRNA expression by RT-PCR. Heparanase protein was detected in both the cytoplasm and the nucleus of heparanase mRNA-positive cells by immunohistochemical staining. Heparanase expression was confirmed in 35 (79.5%) of 44 gastric tumor samples by immunohistochemical staining. However, no or weak heparanase expression was detected in normal gastric mucosa. In situ hybridization showed that the mRNA expression pattern of heparanase was similar to that of the protein, suggesting that increased expression of the heparanase protein at the invasive front was caused by an increase of heparanase mRNA in tumor cells. Analysis of the clinicopathologic features showed stronger heparanase expression in cases of huge growing tumors, extensive invasion to lymph vessels, and regional lymph node metastasis. In gastric cancer, patients with heparanase expression showed significantly poorer prognosis than those without such expression (p = 0.006). In conclusion, our findings suggest that high expression of heparanase in gastric cancer is a strong predictor of poor survival. PMID- 12746472 TI - Telomerase activity and telomerase subunit gene expression levels are not related in prostate cancer: a real-time quantification and in situ hybridization study. AB - Because the mechanisms of telomerase activation in prostate cancer are mainly unknown, we investigated the relationships between telomerase activity and expression levels of human telomerase RNA (hTR) and human telomerase reverse transcriptase (hTERT) mRNA in benign and malignant alterations of the human prostate gland. Using the LightCycler technology, hTERT mRNA expression was quantified in 46 radical prostatectomy and 10 benign prostatic hyperplasia (BPH) cases; hTR expression was quantified in a subset of these tissue samples. Telomerase activity was measured using a quantitative telomeric repeat amplification protocol ELISA assay. Similar to hTR, which was expressed in all tissue samples tested, hTERT mRNA was detected in 98% of the prostate cancer samples and in 30% of the BPH samples. Regarding clinicopathologic variables, telomerase activity was significantly correlated with Gleason score (<7 vs > or =7, p = 0.02). No relationships emerged between normalized hTR or hTERT expression levels and tumor stage, Gleason score, lymph node status, or preoperative serum prostate-specific antigen. Remarkably, one third of all cancer and BPH tissue samples with hTR and hTERT expression lack telomerase activity. Quantitative analyses contradict the assumption that a certain threshold level of hTR or hTERT mRNA is required for telomerase activation, thus indicating that telomerase regulation in prostate cancer occurs more likely on a posttranscriptional level. Nevertheless, the observation that hTR and hTERT mRNA levels are significantly (p < 0.002) correlated suggests some common mechanisms in the up-regulation of hTR and hTERT expression. Because in situ hybridization revealed strong hTERT expression in all cells of the tumor glands but also in high-grade prostatic intraepithelial neoplasia foci, this up-regulation seems to occur early in prostate carcinogenesis. PMID- 12746473 TI - Profile of aberrant CpG island methylation along the multistep pathway of gastric carcinogenesis. AB - To date, several reports on methylation of various genes in gastric cancer (GC) have been published. However, most of these studies focused on cancer tissues or a single gene only and gave no information about the methylation status of specific genes in the premalignant stages or about the concurrent methylation of other genes in specific lesions. We attempted to investigate methylation of multiple genes in a large sample collection of GC (n = 80), gastric adenoma (GA) (n = 79), intestinal metaplasia (IM) (n = 57), and chronic gastritis (CG) (n = 74). We determined the methylation frequency of 12 genes, including APC, COX-2, DAP-kinase, E-cadherin, GSTP1, hMLH1, MGMT, p16, p14, RASSF1A, THBS1, and TIMP3 by methylation-specific PCR. Five different classes of methylation behaviors were found: (1) genes methylated in GC only (GSTP1 and RASSF1A); (2) genes showing low methylation frequency (<12%) in CG, IM, and GA, but significantly higher methylation frequency in GC (COX-2, hMLH1, and p16); (3) a gene with low and similar methylation frequency (8.8-21.3%) in four-step lesions (MGMT); (4) genes with high and similar methylation frequency (53-85%) in four-step lesions (APC and E-cadherin); and (5) genes showing an increasing tendency with or without fluctuation of the methylation frequency along the progression (DAP-kinase, p14, THBS1, and TIMP3). The average number of methylated genes was 2.7, 3.6, 3.4, and 5.2 per 12 tested genes in CG, IM, GA, and GC, respectively. Our results suggest that tumor suppressor genes show a gene type-specific methylation profile and that aberrant CpG island methylation tends to accumulate along the pathway of multistep carcinogenesis. PMID- 12746474 TI - Dysregulation of DNA repair pathways in a transforming growth factor alpha/c-myc transgenic mouse model of accelerated hepatocarcinogenesis. AB - Previous work from our laboratory has implicated oxidative DNA damage and genetic instability in the etiology of transforming growth factor-alpha (TGFalpha)/c-myc associated hepatocarcinogenesis. In contrast, oxidative DNA damage was lower in c myc single-transgenic mice, consistent with less chromosomal damage and with later and more benign tumor formation. We examined whether defects in the DNA repair pathways contribute to the acceleration of liver cancer in TGFalpha/c-myc mice. A cDNA expression array containing 140 known genes and multiplex RT-PCR were used to compare the basal levels of expression of DNA repair genes at the dysplastic stage. Thirty-five percent (8/23) and 43% (10/23) of DNA repair genes were constitutively up-regulated in 10-week-old TGFalpha/c-myc and c-myc transgenic livers, respectively, compared with wild-type controls. The commonly up-regulated genes were OGG1 and NTH1 of base excision repair; ERCC5, RAD23A, and RAD23B of nucleotide excision repair; and RAD50, RAD52, and RAD54 involved in DNA strand break repair. Additional treatment with a peroxisome proliferator, Wy 14,643, known to increase the level of oxidants in the liver, failed to induce a further increase in the expression level of DNA repair enzymes in TGFalpha/c-myc but not in c-myc or wild-type livers. Moreover, expression of several genes, including Ku80, PMS2, and ATM, was decreased in TGFalpha/c-myc livers, suggesting a fault or inefficient activation of the DNA repair pathway upon induction of oxidative stress. Together, the results show that DNA damage response is attenuated in TGFalpha/c-myc mice, creating a condition that may contribute to acceleration of liver cancer in this model. PMID- 12746475 TI - Apoptotic body engulfment by a human stellate cell line is profibrogenic. AB - Hepatocyte apoptosis and stellate cell activation are both features of chronic liver diseases, but a relationship between these events has not been explored. In macrophages, engulfment of apoptotic bodies induces expression of transforming growth factor-beta (TGF-beta), a profibrogenic cytokine. We examined whether a similar response occurs in stellate cells. Fluorescently labeled hepatocyte apoptotic bodies were added to cultures of primary and immortalized human stellate cells. Stellate cells, but not hepatocytes, readily engulfed apoptotic bodies in a time-dependent manner as assessed by confocal microscopy. The activation of primary and immortalized human stellate cells after incubation with apoptotic bodies, as well as their fibrogenic activity, was indicated by an increase in alpha-smooth muscle actin (primary cells), TGF-beta1, and collagen alpha1(I) mRNA (primary and immortalized cells). The profibrogenic response was dependent upon apoptotic body engulfment, because nocodazole, a microtubule inhibiting agent, blocked both the engulfment and the increase of TGF-beta1 and collagen alpha1(I) mRNA. As described in primary rodent stellate cells, up regulation of collagen alpha1(I) mRNA was inhibited by a PI-3K inhibitor (LY294002) and a p38 mitogen-activated protein kinase inhibitor (SB203580) in LX 1 cells. In conclusion, these data support a model in which engulfment of hepatocyte apoptotic bodies by stellate cells leads to a fibrogenic response by eliciting a kinase-signaling pathway. PMID- 12746476 TI - Stromal cell-derived factor-1 from biliary epithelial cells recruits CXCR4 positive cells: implications for inflammatory liver diseases. AB - Although stromal cell-derived factor-1 (SDF-1) plays an important role in hematopoiesis in the fetal liver, the role after birth remains to be clarified. We investigated the role of SDF-1 and its receptor, CXCR4, in 75 patients; this included controls and patients with viral hepatitis, liver cirrhosis, primary biliary cirrhosis, primary sclerosing cholangitis, and autoimmune hepatitis. Interestingly, SDF-1 appeared up-regulated in biliary epithelial cells (BEC) of inflammatory liver disease. Furthermore, in inflammatory liver diseases, SDF-1 was expressed by BEC of interlobular and septal bile ducts and by proliferated bile ductules. The message expression of SDF-1 in BEC was confirmed at a single cell level by RT-PCR and laser capture microdissection. The plasma levels of SDF 1 were significantly higher in patients with liver diseases than in normal controls. Flow cytometric analysis of the surface expression of CXCR4 showed that most liver-infiltrating lymphocytes express CXCR4 and the intensity was up regulated more significantly in liver-infiltrating lymphocytes than in peripheral blood lymphocytes. These results suggest that increased SDF-1 production by BEC may play an important role in the recruitment of CXCR4-positive inflammatory cells into the diseased livers. These data are significant because modulation of the SDF-1/CXCR4 interaction has therapeutic implications for inflammatory liver diseases. PMID- 12746477 TI - Chimerism of murine fetal bone marrow by maternal cells occurs in late gestation and persists into adulthood. AB - Studies of murine severe combined immune-deficient (scid/scid) fetuses gestating in transgene-tagged immune competent dams have established high frequencies of transplacental trafficking of nucleated maternal cells. Maternal cells first appeared in thymus at gestation day (gd) 12.5 and were present in more than 90% of late gestation fetuses. Morphologically heterogeneous maternal cells were located predominantly in bone marrow and thymus and also occasionally in liver, spleen, and nonlymphoid organs. We have now evaluated maternal cell chimerism in offspring with normal lymphoid development. Genetically normal blastocysts from random-bred CD1 mice were transferred to C57BL/6J- lacZ transgene-tagged ROSA26 females. Serial sectioning of fetuses followed by histochemistry for lacZ expressing cells was used to comprehensively define organs containing maternal cells. Fetuses, sectioned in their entirety, had no detectable maternal cells before gd 16.5. Morphologically homogenous, nucleated maternal cells were first present in fetal bone marrow cavities at gd 16.5 and were evident in all offspring in later gestation. Postnatally, maternal cells were also present in bone marrow cavities into adulthood, as determined by lacZ histochemistry and PCR amplification of the maternal transgene. The frequency of maternally derived cells in postnatal bone marrow was increased compared with late gestation, and occasionally, maternal cells were detected in postnatal spleen. The normalcy of maternal cell transfer to genetically immune competent progeny and their long term engraftment is suggestive of a functional role for maternal cells in offspring. PMID- 12746478 TI - Immunopolarization of CD4+ and CD8+ T cells to Type-1-like is associated with melanocyte loss in human vitiligo. AB - Vitiligo is an autoimmune condition characterized by loss of epidermal melanocytes. High frequencies of melanocyte-reactive cytotoxic T cells in the peripheral blood of vitiligo patients and the observed correlation between perilesional T-cell infiltration and melanocyte loss in situ suggest the important role of cellular autoimmunity in the pathogenesis of this disease. We isolated T cells from both perilesional and nonlesional skin biopsies obtained from five vitiligo patients, then cloned and analyzed their profile of cytokine production after short-term, nonspecific expansion in vitro. Perilesional T-cell clones (TCC) derived from patients with vitiligo exhibited a predominant Type-1 like cytokine secretion profile, whereas the degree of Type-1 polarization in uninvolved skin-derived TCC correlated with the process of microscopically observed melanocyte destruction in situ. Detailed analysis of broad spectrum of cytokines produced by perilesional- and nonlesional-derived CD4+ and CD8+ TCC confirmed polarization toward Type-1-like in both CD4 and CD8 compartments, which paralleled depigmentation process observed locally in the skin. Furthermore, CD8+ TCC derived from two patients also were analyzed for reactivity against autologous melanocytes. The antimelanocyte cytotoxic reactivity was observed among CD8+ TCC isolated from perilesional biopsies of two patients with vitiligo. Finally, in two of five patients, tetramer analysis revealed presence of high frequencies of Mart-1-specific CD8 T cells in T-cell lines derived from perilesional skin. Altogether our data support the role of cellular mechanisms playing a significant part in the destruction of melanocytes in human autoimmune vitiligo. PMID- 12746480 TI - Cardiac-restricted ankyrin-repeated protein is differentially induced in duchenne and congenital muscular dystrophy. AB - Cardiac ankyrin-repeated protein (CARP) has been shown to associate with a transcription factor, YB-1, that may activate expression of the ventricular myosin light chain-2 gene during cardiogenesis. CARP is induced in the adult hypertrophic heart subjected to pressure overload, suggesting that CARP may play important functional roles in both embryonic and adult hearts. Although CARP expression was initially believed to be restricted to the heart, we found recently that CARP is induced strongly in human fetal skeletal muscle and in experimentally denervated skeletal muscle, leading us to speculate that CARP may also play important roles in skeletal muscle. In the present study, we found that in rats initially damaged by a single injection of bupivacaine, CARP expression was induced strongly in regenerating muscles with a peak 3 days after the injection, followed by down-regulation to undetectable levels after 28 days. Although CARP was coexpressed with embryonic myosin heavy chain (MHC) in regenerating myofibers, CARP expression persisted even after down-regulation of embryonic MHC expression, whereas it began to decrease before the onset of slow or fast MHC expression, suggesting that CARP is expressed at a specific differentiation stage during muscle regeneration. We analyzed the expression of CARP in muscle biopsy specimens from 14 patients with muscular dystrophy (MD) and detected high expression of CARP in 13 of the 14 cases. CARP-positive myofibers were detected more often in congenital muscular dystrophy (CMD) than in Duchenne muscular dystrophy (DMD). We found that CARP was expressed exclusively, and at a high level, in small regenerating myofibers that express embryonic MHC in DMD, which suggested that CARP could be used as a marker of muscle regeneration in DMD. On the other hand, in CMD, expression of CARP was not limited to regenerating fibers, being detectable in myofibers expressing embryonic MHC and those expressing mature-type MHC. These findings suggest that the differentiation stage of CARP-positive myofibers in DMD and CMD may differ. PMID- 12746479 TI - Alterations of biologic properties and gene expression in nasopharyngeal epithelial cells by the Epstein-Barr virus-encoded latent membrane protein 1. AB - Undifferentiated nasopharyngeal carcinoma (NPC) is closely associated with EBV infection, and the EBV-encoded latent membrane protein 1 (LMP1) is frequently detected in NPC. However, little is known about the pathologic roles of LMP1 in this disease. Recently, we reported the morphologic transformation and increased expression of the LAMC2 and ITGalpha6 genes in LMP1-expressing NPC cell lines. In this study, we further examine the effects of LMP1 in an immortalized nasopharyngeal epithelial cell line called NP69. This cell line was established from primary nonmalignant nasopharyngeal epithelial cells and may represent a model of premalignant nasopharyngeal epithelial cells. LMP1 induced many phenotypic changes in NP69 cells. These include morphologic transformation, increased cell proliferation, anchorage-independent growth, resistance to serum free-induced cell death, and enhanced cell migration and invasion. In addition, expression array analysis identified 28 genes that demonstrated a more than 2 fold difference in expression of NP69 cells expressing LMP1 when compared with a vector control. Two of the up-regulated genes (VEGF and vimentin) identified have been previously reported as LMP1 targets. The majority of the identified genes are associated with cell growth, differentiation, cell shape, and invasion. The present findings support the proposed roles of LMP1 in promoting cell transformation, migration, and invasion in premalignant nasopharyngeal epithelial cells. The present study also indicates the activation of the Ras/MAPK pathway in LMP1-expressing cells, which may be involved in mediating some of the transforming effects of LMP1 observed in nasopharyngeal epithelial cells. PMID- 12746481 TI - Inhibition of cytokine-induced fractalkine production by bacterial invasion of human-dermal fibroblasts. AB - Fractalkine (FKN/CX3CL1) is an atypical chemokine, for which a major biological function has not yet emerged. However, recent data suggest a role in immune responses in the skin. In this study, we analyzed fractalkine (FKN) secretion by human-dermal fibroblasts after exposure to pro-inflammatory cytokines or to invasive and noninvasive strains of Escherichia coli. Incubation of fibroblasts with TNF-alpha and IL-1beta induced a delayed expression of soluble FKN, compared with the rapid secretion of other chemokines including IL-8 (CXCL8), monocyte chemotactic protein-1 (CCL2), and RANTES (regulated upon activation, normal T cell expressed and secreted; CCL5). TNF-alpha and IFN-gamma gamma were more potent at inducing FKN secretion than was IL-1beta. Very little FKN was detected on the cell surface. FKN was not detected after incubation with the bacteria, regardless of the strain used. In contrast, both invasive and noninvasive E. coli triggered the release of IL-8 and monocyte chemotactic protein-1 in a dose response manner, whereas RANTES was produced only in response to the invasive strain. Finally, incubation of fibroblasts with the invasive strain of E. coli inhibited TNF-alpha- and IFN-gamma-induced production of FKN. These results demonstrate for the first time that human-dermal fibroblasts express FKN, and that the characteristics of FKN secretion are distinct from those of other chemokines produced by these cells during immune responses in the dermis. In addition, our data indicate that bacterial invasion of dermal fibroblasts actively modulates FKN expression. PMID- 12746482 TI - Neurokinin-1 receptor expression and its potential effects on tumor growth in human pancreatic cancer. AB - The neurokinin-1 receptor (NK-1R) and its ligand substance P (SP) are involved in the pathogenesis of certain neural tumors. Because nerves are significantly altered in pancreatic cancer, evidence for alteration of this pathway in human pancreatic cancer was sought. Expression of NK-1R was analyzed by real-time quantitative RT-PCR, in situ hybridization, immunohistochemistry, and Western blot analysis in normal human pancreatic and pancreatic cancer tissue samples and in pancreatic cancer cell lines. Furthermore, the influence of SP analogs and of the NK-1R antagonist MEN 11467 on pancreatic cancer cell growth was analyzed by sulforhodamine B (SRB) assay. By real-time quantitative RT-PCR, NK-1R mRNA was increased 36.7-fold (p < 0.001) in human pancreatic cancer samples compared with normal controls. Enhanced NK-1R expression levels were not related to tumor grade but were associated with advanced tumor stage and poorer prognosis. By in situ hybridization and immunohistochemistry, NK-1R mRNA and immunoreactivity were only occasionally weakly present in acinar and ductal cells in the normal pancreas. In contrast, moderate to strong NK-1R mRNA signals and immunoreactivity were present in most cancer cells. By Western blot analysis, NK-1R was increased 26-fold (p < 0.01) in pancreatic cancer samples in comparison to normal controls. NK-1R mRNA was detected in five pancreatic cancer cell lines by real-time quantitative RT PCR, with the highest levels in CAPAN-1 cells and the lowest in ASPC-1 cells. SP analogs stimulated pancreatic cancer cell growth, depending on the NK-1R expression level, and this effect could be blocked by a selective NK-1R antagonist. These findings illustrate that the NK-1R pathway is activated in human pancreatic cancer and has the potential to contribute to cancer cell growth, thus suggesting the existence of a neuro-cancer cell interaction in vivo. PMID- 12746483 TI - Age-associated changes in histology and gene-expression profile in the rat ventral prostate. AB - The incidence of prostate diseases rises dramatically with age in men, yet little is understood of the mechanisms underlying prostatic senescence and its contribution to disease development in the gland. In Noble rats, aging of the ventral prostate (VP) is characterized morphologically by widespread atrophy of acini, increased accumulation of concretions in glandular lumen, infiltration of inflammatory cells, and focal epithelial atypia. We used a cDNA microarray containing 2388 known transcripts, together with the Tyramide Amplification System and t statistics, to identify differentially expressed genes in the VPs of young (3 months old) and old (16 months old) rats. A total of 78 VP genes were found to be differentially expressed by the two groups; in old rats, 65 VP genes (83%) showed reduced expression and 13 genes (17%) showed increased expression compared with young animals. The age-dependent underexpressed genes fell into several functional clusters: those involved in amino-acid metabolism, protein synthesis, protein secretion and degradation, vesicle/membrane trafficking, energy metabolism, signal transduction, spermidine and spermine syntheses, and cellular defense against stress. The overexpressed genes included iduronate 2 sulfatase, HLA class I locus C heavy chain, membrane cofactor protein of the complement system, TRPM-2, cadherin-associated protein-related, and X-CGD. Post hoc analyses confirmed a progressive decline in the expression of ribophorin II and BiP and a gradual increase in the expression of TRPM-2 in rat VPs as animals aged from 3 to 19 months old. In conclusion, the observed widespread declines in expression of genes involved in protein synthesis, protein fidelity maintenance, anabolism, growth inhibition, and energy metabolism, together with increased expression of genes implicated in cell survival in the VPs of senescent rats, may help explain the susceptibility of the prostates of elderly men to development of disease. PMID- 12746484 TI - Rapid identification of differentially expressed genes by combination of SSH and MOS. PMID- 12746485 TI - Pollen tubes exhibit regular periodic membrane trafficking events in the absence of apical extension. AB - The growing pollen tube provides an excellent single cell model system in which to study the mechanisms determining growth regulation, polarity and periodic behaviour. Previously, using FM4-64, we identified periodic movements within the apical vesicle accumulation that were related to the period of oscillatory growth. This suggested a more complex interdependence between membrane traffic, apical extension and periodicity than previously thought. To investigate this a comparison was made between normally growing and Brefeldin-A-treated, non growing, tubes. Brefeldin-A treatment established an intriguing, stable yet dynamic system of membrane aggregations in the pollen tube tip that exhibited regular movements of material with a 5-7 second period compared with the normal approximately 30 second periodicity observed in growing tubes. Heat treatment was found to reduce period length in both cases. After BFA treatment membrane was demonstrated to flow from the extreme pollen tube apex back through a distinct subapical Brefeldin-A-induced membrane accumulation. The effects of Brefeldin-A on the distribution of ER- and Golgi-targeted fluorescent proteins revealed that ER did not contribute directly to the system of membrane aggregations while only certain compartments of the Golgi might be involved. The involvement of membrane derived from the apical vesicle accumulation was strongly implicated. Calcium measurements revealed that Brefeldin-A abolished the typical tip-focused calcium gradient associated with growth and there were no obvious periodic fluctuations in apical calcium associated with the continued periodic Brefeldin-A membrane aggregation associated movements. Our experiments reveal an underlying periodicity in the pollen tube that is independent of secretion, apical extension and the oscillating tip-focused calcium gradient normally associated with growth, but requires an active actin cytoskeleton. PMID- 12746486 TI - A 48 kDa integral membrane phosphoprotein orchestrates the cytoskeletal dynamics that generate amoeboid cell motility in Ascaris sperm. AB - Protrusion of the lamellipod in the crawling sperm of Ascaris is tightly coupled to the localized vectorial assembly and bundling of the major sperm protein cytoskeleton. In cell-free extracts of sperm, vesicles derived from the leading edge membrane reconstitute protrusion by directing the assembly of columnar meshworks of major sperm protein filaments that push the vesicle forward as they elongate. Treatment with proteases or a tyrosine phosphatase abolished vesicle activity, suggesting the involvement of a membrane phosphoprotein. Fractionation of vesicle proteins by sequential detergent lysis, size exclusion chromatography and immunoprecipitation with antiphosphotyrosine antibody identified a 48 kDa integral membrane phosphoprotein as the only sperm membrane component required to nucleate major sperm protein polymerization under physiological conditions. Immunolabeling assays showed that this protein is distributed uniformly in the sperm plasma membrane, but that its active phosphorylated form is located only at sites of major sperm protein polymerization at the leading edge. Because this protein specifies sites of cytoskeletal assembly, we have named it major sperm protein polymerization organizing protein (MPOP). The phosphorylation of MPOP is pH sensitive and appears to require a soluble tyrosine kinase. Comparison of the activity of MPOP to that of analogous membrane proteins in actin-based systems emphasizes the importance of precise transmission of information from the membrane to the cytoskeleton in amoeboid cell motility. PMID- 12746487 TI - CD105 prevents apoptosis in hypoxic endothelial cells. AB - CD105, a marker of endothelial cells, is abundantly expressed in tissues undergoing angiogenesis and is a receptor for transforming growth factorbeta. The pivotal role of CD105 in the vascular system was demonstrated by the severe vascular defects that occur in CD105-knockout mice, but the exact mechanisms for CD105 regulation of vascular development have not been fully elucidated. In light of the function of CD105 and the importance of hypoxia in neovascularisation, we speculated that CD105 is involved in hypoxia-initiated angiogenesis. Using tissue cultured human microvascular endothelial cells, we have investigated the effects of hypoxic stress on CD105 gene expression. Hypoxia induced a significant increase in membrane-bound and secreted CD105 protein levels. CD105 mRNA and promoter activity were also markedly elevated, the latter returning to the basal level after 16 hours of hypoxic stress. Hypoxia induced cell cycle arrest at the G0/G1 phases and massive cell apoptosis after 24 hours through a reduction in the Bcl-2 to Bax ratio, downregulation of Bcl-XL and Mcl-1, and upregulation of caspase-3 and caspase-8. The consequence of CD105 upregulation was revealed using an antisense approach and a TUNEL assay. Suppression of CD105 increased cell apoptosis under hypoxic stress in the absence of TGFbeta1. Furthermore, hypoxia and TGFbeta1 synergistically induced apoptosis in the CD105-deficient cells but not in the control cells. We conclude that hypoxia is a potent stimulus for CD105 gene expression in vascular endothelial cells, which in turn attenuates cell apoptosis and thus contributes to angiogenesis. PMID- 12746488 TI - S100A13 mediates the copper-dependent stress-induced release of IL-1alpha from both human U937 and murine NIH 3T3 cells. AB - Copper is involved in the promotion of angiogenic and inflammatory events in vivo and, although recent clinical data has demonstrated the potential of Cu2+ chelators for the treatment of cancer in man, the mechanism for this activity remains unknown. We have previously demonstrated that the signal peptide-less angiogenic polypeptide, FGF1, uses intracellular Cu2+ to facilitate the formation of a multiprotein aggregate that enables the release of FGF1 in response to stress and that the expression of the precursor form but not the mature form of IL-1alpha represses the stress-induced export of FGF1 from NIH 3T3 cells. We report here that IL-1alpha is a Cu2+-binding protein and human U937 cells, like NIH 3T3 cells, release IL-1alpha in response to temperature stress in a Cu2+ dependent manner. We also report that the stress-induced export of IL-1alpha involves the intracellular association with the Cu2+-binding protein, S100A13. In addition, the expression of a S100A13 mutant lacking a sequence novel to this gene product functions as a dominant-negative repressor of IL-1alpha release, whereas the expression of wild-type S100A13 functions to eliminate the requirement for stress-induced transcription. Lastly, we present biophysical evidence that IL-1alpha may be endowed with molten globule character, which may facilitate its release through the plasma membrane. Because Cu2+ chelation also represses the release of FGF1, the ability of Cu2+ chelators to potentially serve as effective clinical anti-cancer agents may be related to their ability to limit the export of these proinflammatory and angiogenic signal peptide-less polypeptides into the extracellular compartment. PMID- 12746489 TI - Regulation of EDEN-dependent deadenylation of Aurora A/Eg2-derived mRNA via phosphorylation and dephosphorylation in Xenopus laevis egg extracts. AB - Deadenylation is an intimate part of the post-transcriptional regulation of maternal mRNAs in embryos. EDEN-BP is so far the only known member of a complex regulating the deadenylation of maternal mRNA in Xenopus laevis embryos in a manner that is dependent on the 3'-untranslated region called EDEN (embryo deadenylation element). In this report, we show that calcium activation of cell free extracts triggers EDEN binding protein (EDEN-BP) dephosphorylation and concomitant deadenylation of a chimeric RNA bearing Aurora A/Eg2 EDEN sequence. Deadenylation of mRNA deprived of EDEN sequence (default deadenylation) does not change with egg activation. Kinase and phosphatase inhibitors downregulate EDEN dependent deadenylation but they do not substantially influence default deadenylation. Using indestructible Delta90 cyclin B to revert interphase extracts to the M-phase, we show that modulation of EDEN-dependent deadenylation is independent of M-phase promoting factor (MPF) activity. These results suggest that the increase in EDEN-dependent deadenylation following egg activation is achieved, at least partially, via dephosphorylation and/or phosphorylation of regulatory proteins, including EDEN-BP dephosphorylation. This regulation proceeds in a manner independent from MPF inactivation. PMID- 12746490 TI - Glutamate-mediated [Ca2+]c dynamics in spontaneously firing dopamine neurons of the rat substantia nigra pars compacta. AB - The mechanism by which glutamate regulates the cytosolic free Ca2+ concentration ([Ca2+]c) in spontaneously firing dopamine neurons is not clear. Thus we have investigated the glutamate-mediated [Ca2+]c dynamics in the acutely isolated dopamine neurons from the rat substantia nigra pars compacta by measuring [Ca2+]c and spontaneously occurring action potentials (SAPs). The freshly isolated dopamine neurons showed tetrodotoxin (TTX)-sensitive spontaneous firing of 2-3 Hz and the resting [Ca2+]c decreased with abolition of the SAPs. The level of [Ca2+]c was affected by the spontaneous firing rate. In the presence of the Na+ channel antagonist, TTX (0.5 microM), glutamate increased [Ca2+]c by activating different glutamate receptors depending on the glutamate concentration used. Addition of glutamate at low concentrations (<3 microM) raised [Ca2+]c mainly by activating metabotropic glutamate receptors (mGluR), whereas at high concentrations (>10 microM) it raised [Ca2+]c mainly by activating AMPA/kainate receptors. The contribution of NMDA receptors to the glutamate-mediated [Ca2+]c rises was largest at intermediate concentrations of glutamate. Activation of mGluR elicited a Ca2+ release from intracellular Ca2+ stores and continuous Ca2+ influx out of the cell. The spontaneous firing activities were highly enhanced by submicromolar levels of glutamate and abolished at levels above 10 microM. From these results, we conclude that at low glutamate concentrations the [Ca2+]c in the dopamine neurons is mainly governed by mGluR and the firing activities, whose rate is regulated at submicromolar glutamate concentrations, but at higher glutamate concentrations [Ca2+]c is dominantly affected by AMPA/kainate receptors. PMID- 12746491 TI - Centrin deficiency in Chlamydomonas causes defects in basal body replication, segregation and maturation. AB - Centrin, a 20 kDa calcium-binding protein, is a constituent of contractile basal body-associated fibers in protists and of various centrosomal structures. A construct inducing centrin RNAi was used to study the effect of centrin deficiency in Chlamydomonas. Transformants contained variable amounts of residual centrin (down to 5% of wild-type) and lacked centrin fibers. They displayed a variable flagellar number phenotype with mostly nonflagellate cells, suggesting that centrin is required for basal body assembly. Furthermore, basal bodies often failed to dock to the plasma membrane and to assemble flagella, and displayed defects in the flagellar root system indicating that centrin deficiency interferes with basal body development. Multiple basal bodies caused the formation of additional microtubular asters, whereas the microtubular cytoskeleton was disordered in most cells without basal bodies. The number of multinucleated cells was increased, indicating that aberrant numbers of basal bodies interfered with the cytokinesis of Chlamydomonas. In contrast to wild-type cells, basal bodies in centrin-RNAi cells were separated from the spindle poles, suggesting a role of centrin in tethering basal bodies to the spindle. To test whether an association with the spindle poles is required for correct basal body segregation, we disrupted centrin fibers in wild-type cells by over-expressing a nonfunctional centrin-GFP. In these cells, basal bodies were disconnected from the spindle but segregation errors were not observed. We propose that basal body segregation in Chlamydomonas depends on an extranuclear array of microtubules independent of the mitotic spindle. PMID- 12746492 TI - Unexpected roles of a Dictyostelium homologue of eukaryotic EF-2 in growth and differentiation. AB - EF-2 is believed to be indispensable for polypeptide chain elongation in protein synthesis and therefore for cell proliferation. Surprisingly, we could isolate ef2 null cells from Dictyostelium discoideum that exhibited almost normal growth and protein synthesis, which suggests that there is another molecule capable of compensating for EF-2 function. The knock-out of Dictyostelium EF-2 (Dd-EF2H; 101 kDa phosphoprotein) impairs cytokinesis, resulting in formation of multinucleate cells. The initiation of differentiation, including the acquisition of aggregation competence, was delayed in Dd-ef2 null cells compared with that in wild-type. By contrast, Dd-ef2 overexpression enhanced the progression of differentiation, thus indicating a positive involvement of Dd-EF2H in growth/differentiation transition. PMID- 12746493 TI - Functional divergence within the APETALA3/PISTILLATA floral homeotic gene lineages. AB - Changes in homeotic gene expression patterns or in the functions of the encoded proteins are thought to play a prominent role in the evolution of new morphologies. The floral homeotic APETALA3 (AP3) and PISTILLATA (PI) genes encode MADS domain-containing transcription factors required to specify petal and stamen identities in Arabidopsis. We have previously shown that perianth expression of AP3 and PI homologs varies in different groups of angiosperms with diverse floral structures, suggesting that changes in expression may contribute to changing morphology. We have investigated the possibility that changes in the functions of the encoded gene products may also have played a role in the evolution of different floral morphologies. AP3 and PI are members of paralogous gene lineages and share extensive similarity along the length of the protein products. Genes within these lineages encode products with characteristic C-terminal motifs that we show are critical for functional specificity. In particular, the C terminus of AP3 is sufficient to confer AP3 functionality on the heterologous PI protein. Furthermore, we have shown that the evolution of the divergent AP3 C-terminal domain in the core eudicots is correlated with the acquisition of a role in specifying perianth structures. These results suggest that divergence in these sequence motifs has contributed to the evolution of distinct functions for these floral homeotic gene products. PMID- 12746494 TI - Global atmospheric black carbon inferred from AERONET. AB - AERONET, a network of well calibrated sunphotometers, provides data on aerosol optical depth and absorption optical depth at >250 sites around the world. The spectral range of AERONET allows discrimination between constituents that absorb most strongly in the UV region, such as soil dust and organic carbon, and the more ubiquitously absorbing black carbon (BC). AERONET locations, primarily continental, are not representative of the global mean, but they can be used to calibrate global aerosol climatologies produced by tracer transport models. We find that the amount of BC in current climatologies must be increased by a factor of 2-4 to yield best agreement with AERONET, in the approximation in which BC is externally mixed with other aerosols. The inferred climate forcing by BC, regardless of whether it is internally or externally mixed, is approximately 1 W/m2, most of which is probably anthropogenic. This positive forcing (warming) by BC must substantially counterbalance cooling by anthropogenic reflective aerosols. Thus, especially if reflective aerosols such as sulfates are reduced, it is important to reduce BC to minimize global warming. PMID- 12746496 TI - Interaction of soft condensed materials with living cells: phenotype/transcriptome correlations for the hydrophobic effect. AB - The assessment of biomaterial compatibility relies heavily on the analysis of macroscopic cellular responses to material interaction. However, new technologies have become available that permit a more profound understanding of the molecular basis of cell-biomaterial interaction. Here, both conventional phenotypic and contemporary transcriptomic (DNA microarray-based) analysis techniques were combined to examine the interaction of cells with a homologous series of copolymer films that subtly vary in terms of surface hydrophobicity. More specifically, we used differing combinations of N-isopropylacrylamide, which is presently used as an adaptive cell culture substrate, and the more hydrophobic, yet structurally similar, monomer N-tert-butylacrylamide. We show here that even discrete modifications with respect to the physiochemistry of soft amorphous materials can lead to significant impacts on the phenotype of interacting cells. Furthermore, we have elucidated putative links between phenotypic responses to cell-biomaterial interaction and global gene expression profile alterations. This case study indicates that high-throughput analysis of gene expression not only can greatly refine our knowledge of cell-biomaterial interaction, but also can yield novel biomarkers for potential use in biocompatibility assessment. PMID- 12746495 TI - Architecture of initiation-competent 12-subunit RNA polymerase II. AB - RNA polymerase (Pol) II consists of a 10-polypeptide catalytic core and the two subunit Rpb4/7 complex that is required for transcription initiation. Previous structures of the Pol II core revealed a "clamp," which binds the DNA template strand via three "switch regions," and a flexible "linker" to the C-terminal repeat domain (CTD). Here we derived a model of the complete Pol II by fitting structures of the core and Rpb4/7 to a 4.2-A crystallographic electron density map. Rpb4/7 protrudes from the polymerase "upstream face," on which initiation factors assemble for promoter DNA loading. Rpb7 forms a wedge between the clamp and the linker, restricting the clamp to a closed position. The wedge allosterically prevents entry of the promoter DNA duplex into the active center cleft and induces in two switch regions a conformation poised for template-strand binding. Interaction of Rpb4/7 with the linker explains Rpb4-mediated recruitment of the CTD phosphatase to the CTD during Pol II recycling. The core-Rpb7 interaction and some functions of Rpb4/7 are apparently conserved in all eukaryotic and archaeal RNA polymerases but not in the bacterial enzyme. PMID- 12746498 TI - Complete, 12-subunit RNA polymerase II at 4.1-A resolution: implications for the initiation of transcription. AB - The x-ray structure of complete RNA polymerase II from Saccharomyces cerevisiae has been determined, including a heterodimer of subunits Rpb4 and Rpb7 not present in previous "core" polymerase II structures. The heterodimer maintains the polymerase in the conformation of a transcribing complex, may bind RNA as it emerges from the enzyme, and is in a position to interact with general transcription factors and the Mediator of transcriptional regulation. PMID- 12746497 TI - Asialoerythropoietin is a nonerythropoietic cytokine with broad neuroprotective activity in vivo. AB - Erythropoietin (EPO) is a tissue-protective cytokine preventing vascular spasm, apoptosis, and inflammatory responses. Although best known for its role in hematopoietic lineages, EPO also affects other tissues, including those of the nervous system. Enthusiasm for recombinant human erythropoietin (rhEPO) as a potential neuroprotective therapeutic must be tempered, however, by the knowledge it also enlarges circulating red cell mass and increases platelet aggregability. Here we examined whether erythropoietic and tissue-protective activities of rhEPO might be dissociated by a variation of the molecule. We demonstrate that asialoerythropoietin (asialoEPO), generated by total enzymatic desialylation of rhEPO, possesses a very short plasma half-life and is fully neuroprotective. In marked contrast with rhEPO, this molecule at doses and frequencies at which rhEPO exhibited erythropoiesis, did not increase the hematocrit of mice or rats. AsialoEPO appeared promptly within the cerebrospinal fluid after i.v. administration; intravenously administered radioiodine-labeled asialoEPO bound to neurons within the hippocampus and cortex in a pattern corresponding to the distribution of the EPO receptor. Most importantly, asialoEPO exhibits a broad spectrum of neuroprotective activities, as demonstrated in models of cerebral ischemia, spinal cord compression, and sciatic nerve crush. These data suggest that nonerythropoietic variants of rhEPO can cross the blood-brain barrier and provide neuroprotection. PMID- 12746499 TI - Chloride, not sodium, stimulates expression of the gamma subunit of Na/K-ATPase and activates JNK in response to hypertonicity in mouse IMCD3 cells. AB - Hypertonicity induced by NaCl, but not by urea or mannitol, up-regulates expression of the gamma subunit of Na/K-ATPase in cells of the murine inner medullary collecting duct line (IMCD3) by activation of the Jun kinase 2 (JNK2) pathways. We examined the ionic mediators of the osmosensitive response. An increase in osmolality to 550 milliosmoles per kg of water (mosmol/kgH2O) for 48 h by replacement of NaCl with choline chloride did not prevent the up-regulation of the gamma subunit. Neither Na+ ionophores nor inhibitors of cellular Na+ uptake altered the up-regulation of the gamma subunit or JNK activation. Changes in cell cation concentrations driven by incubation in low-K+ medium were effective in up-regulating the alpha1 subunit of Na/K-ATPase but did not have any effect on the gamma subunit. The replacement of NaCl with choline chloride did not down-regulate gamma-subunit expression in cells adapted to hypertonicity. In contrast, the replacement of NaCl with sodium acetate, or pretreatment of cells with the Cl- channel inhibitor 5-nitro-2-(3-phenylpropyl-amino)benzoic acid (NPPB) completely blocked gamma-subunit up-regulation, inhibited JNK activation, and caused a significant decrement in cell survival in hypertonic but not isotonic conditions. In adapted cells, replacement of 300 mosmol/kgH2O NaCl with sodium acetate resulted in down-regulation of the gamma subunit. In conclusion, we describe a Na+-independent, Cl--dependent mechanism for hypertonicity-mediated activation of the JNK and the subsequent synthesis of the gamma subunit of Na/K ATPase, which are necessary for cellular survival in these anisotonic conditions. PMID- 12746500 TI - Specificity of short interfering RNA determined through gene expression signatures. AB - Short interfering RNA (siRNA) is widely used for studying gene function and holds great promise as a tool for validating drug targets and treating disease. A critical assumption in these applications is that the effect of siRNA on cells is specific, i.e., limited to the specific knockdown of the target gene. In this article, we characterize the specificity of siRNA by applying gene expression profiling. Several siRNAs were designed against different regions of the same target gene for three different targets. Their effects on cells were compared by using DNA microarrays to generate gene expression signatures. When the siRNA design and transfection conditions were optimized, the signatures for different siRNAs against the same target were shown to correlate very closely, whereas the signatures for different genes revealed no correlation. These results indicate that siRNA is a highly specific tool for targeted gene knockdown, establishing siRNA-mediated gene silencing as a reliable approach for large-scale screening of gene function and drug target validation. PMID- 12746503 TI - Plant Physiology's Best Paper Award 2002. PMID- 12746501 TI - Myocardial recovery from ischemia is impaired in CD36-null mice and restored by myocyte CD36 expression or medium-chain fatty acids. AB - Long-chain fatty acid uptake, which provides a large part of myocardial energy, is impaired in human and murine hearts deficient in the membrane fatty acid translocase, FAT/CD36. We examined myocardial function in CD36-null mice using the working heart. Fatty acid oxidation and stores of glycogen, triglycerides, and ATP were reduced in CD36-deficient hearts and were restored to WT levels by rescue of myocyte CD36. Under normal perfusion conditions, CD36-null hearts had similar cardiac outputs and end-diastolic pressures as WT or transgenic hearts. After 6 min of ischemia, cardiac output decreased by 41% and end diastolic pressure tripled for CD36-null hearts, with no significant changes in WT or transgenic hearts. Null hearts also failed more frequently after ischemia as compared with WT or transgenics. To dissect out contribution of fatty acid uptake, a perfusate-lacking fatty acids was used. This decreased cardiac output after ischemia by 30% in WT hearts as compared with 50% for CD36-deficient hearts. End diastolic pressure, a negative index of myocardial performance, increased after ischemia in all heart types. Addition to the perfusate of a medium-chain fatty acid (caprylic acid) that does not require CD36 for uptake alleviated poor ischemic tolerance of CD36-null hearts. In summary, recovery from ischemia is compromised in CD36-deficient hearts and can be restored by CD36 rescue or by supplying medium-chain fatty acids. It would be important to determine whether the findings apply to the human situation where polymorphisms of the CD36 gene are relatively common. PMID- 12746504 TI - Agricultural ethics. PMID- 12746505 TI - Value judgments and risk comparisons. The case of genetically engineered crops. PMID- 12746502 TI - Beta3 integrin deficiency promotes atherosclerosis and pulmonary inflammation in high-fat-fed, hyperlipidemic mice. AB - Hyperlipidemia promotes the chronic inflammatory disease atherosclerosis through poorly understood mechanisms. Atherogenic lipoproteins activate platelets, but it is unknown whether platelets contribute to early inflammatory atherosclerotic lesions. To address the role of platelet aggregation in diet-induced vascular disease, we studied beta3 integrin-deficient mice (lacking platelet integrin alphaIIbbeta3 and the widely expressed nonplatelet integrin alphavbeta3) in two models of atherosclerosis, apolipoprotein E (apoE)-null and low-density lipoprotein receptor (LDLR)-null mice. Unexpectedly, a high-fat, Western-type (but not a low-fat) diet caused death in two-thirds of the beta3-/-apoE-/- and half of the beta3-/-LDLR-/- mice due to noninfectious pneumonitis. In animals from both models surviving high-fat feeding, pneumonitis was absent, but aortic atherosclerosis was 2- to 6-fold greater in beta3-/- compared with beta+/+ littermates. Expression of CD36, CD40L, and CD40 was increased in lungs of beta3 /-LDLR-/- mice. Each was also increased in smooth muscle cells cultured from beta3-deficient mice and suppressed by retroviral reconstitution of beta3. These data show that the platelet defect caused by alphaIIbbeta3 deficiency does not impair atherosclerotic lesion initiation. They also suggest that alphavbeta3 has a suppressive effect on inflammation, the loss of which induces atherogenic mediators that are amplified by diet-induced hyperlipidemia. PMID- 12746506 TI - Sandbox ethics in science: sharing of data and materials in plant biology. PMID- 12746507 TI - Sharing publication-related data and materials: responsibilities of authorship in the life sciences. PMID- 12746508 TI - Laser capture microdissection of cells from plant tissues. AB - Laser capture microdissection (LCM) is a technique by which individual cells can be harvested from tissue sections while they are viewed under the microscope, by tacking selected cells to an adhesive film with a laser beam. Harvested cells can provide DNA, RNA, and protein for the profiling of genomic characteristics, gene expression, and protein spectra from individual cell types. We have optimized LCM for a variety of plant tissues and species, permitting the harvesting of cells from paraffin sections that maintain histological detail. We show that RNA can be extracted from LCM-harvested plant cells in amount and quality that are sufficient for the comparison of RNAs among individual cell types. The linear amplification of LCM-captured RNA should permit the expression profiling of plant cell types. PMID- 12746509 TI - Genetic modification removes an immunodominant allergen from soybean. AB - The increasing use of soybean (Glycine max) products in processed foods poses a potential threat to soybean-sensitive food-allergic individuals. In vitro assays on soybean seed proteins with sera from soybean-sensitive individuals have immunoglobulin E reactivity to abundant storage proteins and a few less-abundant seed proteins. One of these low abundance proteins, Gly m Bd 30 K, also referred to as P34, is in fact a major (i.e. immunodominant) soybean allergen. Although a member of the papain protease superfamily, Gly m Bd 30 K has a glycine in the conserved catalytic cysteine position found in all other cysteine proteases. Transgene-induced gene silencing was used to prevent the accumulation of Gly m Bd 30 K protein in soybean seeds. The Gly m Bd 30 K-silenced plants and their seeds lacked any compositional, developmental, structural, or ultrastructural phenotypic differences when compared with control plants. Proteomic analysis of extracts from transgenic seed detected the suppression of Gly m Bd 30 K-related peptides but no other significant changes in polypeptide pattern. The lack of a collateral alteration of any other seed protein in the Gly m Bd 30 K-silenced seeds supports the presumption that the protein does not have a role in seed protein processing and maturation. These data provide evidence for substantial equivalence of composition of transgenic and non-transgenic seed eliminating one of the dominant allergens of soybean seeds. PMID- 12746510 TI - Root exudation and rhizosphere biology. PMID- 12746511 TI - CACTA transposons in Triticeae. A diverse family of high-copy repetitive elements. AB - In comparison with retrotransposons, which comprise the majority of the Triticeae genomes, very few class 2 transposons have been described in these genomes. Based on the recent discovery of a local accumulation of CACTA elements at the Glu-A3 loci in the two wheat species Triticum monococcum and Triticum durum, we performed a database search for additional such elements in Triticeae spp. A combination of BLAST search and dot-plot analysis of publicly available Triticeae sequences led to the identification of 41 CACTA elements. Only seven of them encode a protein similar to known transposases, whereas the other 34 are considered to be deletion derivatives. A detailed characterization of the identified elements allowed a further classification into seven subgroups. The major subgroup, designated the "Caspar " family, was shown by hybridization to be present in at least 3,000 copies in the T. monococcum genome. The close association of numerous CACTA elements with genes and the identification of several similar elements in sorghum (Sorghum bicolor) and rice (Oryza sativa) led to the conclusion that CACTA elements contribute significantly to genome size and to organization and evolution of grass genomes. PMID- 12746512 TI - Expression profiling and bioinformatic analyses of a novel stress-regulated multispanning transmembrane protein family from cereals and Arabidopsis. AB - Cold acclimation is a multigenic trait that allows hardy plants to develop efficient tolerance mechanisms needed for winter survival. To determine the genetic nature of these mechanisms, several cold-responsive genes of unknown function were identified from cold-acclimated wheat (Triticum aestivum). To identify the putative functions and structural features of these new genes, integrated genomic approaches of data mining, expression profiling, and bioinformatic predictions were used. The analyses revealed that one of these genes is a member of a small family that encodes two distinct groups of multispanning transmembrane proteins. The cold-regulated (COR)413-plasma membrane and COR413-thylakoid membrane groups are potentially targeted to the plasma membrane and thylakoid membrane, respectively. Further sequence analysis of the two groups from different plant species revealed the presence of a highly conserved phosphorylation site and a glycosylphosphatidylinositol-anchoring site at the C-terminal end. No homologous sequences were found in other organisms suggesting that this family is specific to the plant kingdom. Intraspecies and interspecies comparative gene expression profiling shows that the expression of this gene family is correlated with the development of freezing tolerance in cereals and Arabidopsis. In addition, several members of the family are regulated by water stress, light, and abscisic acid. Structure predictions and comparative genome analyses allow us to propose that the cor413 genes encode putative G protein-coupled receptors. PMID- 12746514 TI - Mining for single nucleotide polymorphisms and insertions/deletions in maize expressed sequence tag data. AB - We have developed a computer based method to identify candidate single nucleotide polymorphisms (SNPs) and small insertions/deletions from expressed sequence tag data. Using a redundancy-based approach, valid SNPs are distinguished from erroneous sequence by their representation multiple times in an alignment of sequence reads. A second measure of validity was also calculated based on the cosegregation of the SNP pattern between multiple SNP loci in an alignment. The utility of this method was demonstrated by applying it to 102,551 maize (Zea mays) expressed sequence tag sequences. A total of 14,832 candidate polymorphisms were identified with an SNP redundancy score of two or greater. Segregation of these SNPs with haplotype indicates that candidate SNPs with high redundancy and cosegregation confidence scores are likely to represent true SNPs. This was confirmed by validation of 264 candidate SNPs from 27 loci, with a range of redundancy and cosegregation scores, in four inbred maize lines. The SNP transition/transversion ratio and insertion/deletion size frequencies correspond to those observed by direct sequencing methods of SNP discovery and suggest that the majority of predicted SNPs and insertion/deletions identified using this approach represent true genetic variation in maize. PMID- 12746513 TI - In silico identification of putative regulatory sequence elements in the 5' untranslated region of genes that are expressed during male gametogenesis. AB - During pollen development, transcription of a large number of genes results in the appearance of distinct sets of transcripts. Similar mRNA sets are present in pollen of both mono- and dicotyledonous plant species, which indicates an evolutionary conservation of genetic programs that determine pollen gene expression. In pollen, regulation of gene expression occurs at the transcriptional and posttranscriptional level. The 5'-untranslated region (UTR) of several pollen transcripts has been shown to be important for regulation of pollen gene expression. The important regulatory role of 5'-UTR sequences and the evolutionary conservation of genetic programs in pollen led to the hypothesis that the 5'-UTRs of pollen-expressed genes share regulatory sequence elements. In an attempt to identify these pollen 5'-UTR elements, a statistical analysis was performed using 5'-UTR sequences of pollen- and sporophytic-expressed genes. The analysis revealed the presence of several pollen-specific 5'-UTR sequence elements. Assembly of the pollen 5'-UTR elements led to the identification of various consensus sequences, including those that previously have been demonstrated to play a role in the regulation of pollen gene expression. Several pollen 5'-UTR elements were found to be preferentially associated to genes from dicots, wet-type stigma plants, or plants containing bicellular pollen. Moreover, three sequence elements exhibited a preferential association to the 5'-UTR of pollen-expressed genes from Arabidopsis and Brassica napus. Functional implications of these observations are discussed. PMID- 12746515 TI - A role for phosphatidylinositol 3-phosphate in abscisic acid-induced reactive oxygen species generation in guard cells. AB - Guard cells generate reactive oxygen species (ROS) in response to abscisic acid (ABA), which leads to stomatal closing. The upstream steps of the ABA-induced ROS generation pathway remain largely unknown. In animal cells, ROS generation in neutrophils is activated by phosphatidylinositol 3-phosphate (PI3P). Stomatal guard cells contain PI3P and PI 3-kinase activity. In this study, we tested whether PI3P has a role in ROS generation in guard cells exposed to ABA. We found that PI 3-kinase inhibitors wortmannin or LY294002 inhibited ABA-induced ROS generation and stomatal closing. Endosome-binding domain (of human EEA1), which specifically binds to PI3P, also inhibited ABA-induced ROS generation and stomatal closing when overexpressed in guard cells. Hydrogen peroxide partially reversed the effects of wortmannin or LY294002 on ABA-induced stomatal closing. These results support a role for PI3P in ABA-induced ROS generation and stomatal closing movement. PMID- 12746516 TI - Cell death in the unicellular chlorophyte Dunaliella tertiolecta. A hypothesis on the evolution of apoptosis in higher plants and metazoans. AB - Apoptosis is essential for normal growth and development of multicellular organisms, including metazoans and higher plants. Although cell death processes have been reported in unicellular organisms, key elements of apoptotic pathways have not been identified. Here, we show that when placed in darkness, the unicellular chlorophyte alga Dunaliella tertiolecta undergoes a form of cell death reminiscent of apoptosis in metazoans. Many morphological criteria of apoptotic cell death were met, including an increase in chromatin margination, degradation of the nucleus, and DNA fragmentation. Biochemical assays of the activities of cell death-associated proteases, caspases, measured using highly specific fluorogenic substrates, increased with time in darkness and paralleled the morphological changes. The caspase-like activities were inhibited by caspase specific inhibitors. Antibodies raised against mammalian caspases cross-reacted with specific proteins in the alga. The pattern of expression of these immunologically reactive proteins was correlated with the onset of cell death. The occurrence of key components of apoptosis, and particularly a caspase mediated cell death cascade in a relatively ancient linage of eukaryotic photoautotrophs, argues against current theories that cell death evolved in multicellular organisms. We hypothesize that key elements of cell death pathways were transferred to the nuclear genome of early eukaryotes through ancient viral infections in the Precambrian Ocean before the evolution of multicellular organisms and were subsequently appropriated in both metazoan and higher plant lineages. PMID- 12746517 TI - Overexpression of a knotted-like homeobox gene of potato alters vegetative development by decreasing gibberellin accumulation. AB - Potato (Solanum tuberosum) homeobox 1 (POTH1) is a class I homeobox gene isolated from an early-stage tuber cDNA library. The RNA expression pattern of POTH1, unlike that of most other class I knotted-like homeobox genes, is widespread in the cells of both indeterminate and differentiated tissues. Using in situ hybridization, POTH1 transcripts were detected in meristematic cells, leaf primordia, and the vascular procambium of the young stem. Overexpression of POTH1 produced dwarf plants with altered leaf morphology. Leaves were reduced in size and displayed a "mouse-ear" phenotype. The mid-vein was less prominent, resulting in a palmate venation pattern. The overall plant height of overexpression lines was reduced due to a decrease in internode length. Levels of intermediates in the gibberellin (GA) biosynthetic pathway were altered, and the bioactive GA, GA(1), was reduced by one-half in sense mutants. Accumulation of mRNA for GA 20 oxidase1, a key biosynthetic enzyme, decreased in overexpression lines. In vitro tuberization was enhanced under both short- and long-day photoperiods in several POTH1 overexpression lines. Sense lines produced more tubers at a faster rate than controls. These results imply that POTH1 mediates the development of potato by acting as a negative regulator of GA biosynthesis. PMID- 12746519 TI - STA11, a Chlamydomonas reinhardtii locus required for normal starch granule biogenesis, encodes disproportionating enzyme. Further evidence for a function of alpha-1,4 glucanotransferases during starch granule biosynthesis in green algae. AB - In Chlamydomonas reinhardtii, the presence of a defective STA11 locus results in significantly reduced granular starch deposition displaying major modifications in shape and structure. This defect simultaneously leads to the accumulation of linear malto-oligosaccharides (MOS). The mutants of STA11 were showed to lack D enzyme, a plant alpha-1,4 glucanotransferase analogous to the Escherichia coli amylomaltase. We have cloned and characterized both the cDNA and gDNA corresponding to the C. reinhardtii D-enzyme. We now report allele-specific modifications of the D-enzyme gene in the mutants of STA11. These allele-specific modifications cosegregate with the corresponding sta11 mutations, thereby demonstrating that STA11 encodes D-enzyme. MOS production and starch accumulation were investigated during day and night cycles in wild-type and mutant C. reinhardtii cells. We demonstrate that in the algae MOS are produced during starch biosynthesis and degraded during the phases of net polysaccharide catabolism. PMID- 12746518 TI - Clustering of microarray data reveals transcript patterns associated with somatic embryogenesis in soybean. AB - Globular somatic embryos can be induced from immature cotyledons of soybean (Glycine max L. Merr. cv Jack) placed on high levels of the auxin 2,4 dichlorophenoxyacetic acid (2,4-D). Somatic embryos develop from the adaxial side of the cotyledon, whereas the abaxial side evolves into a callus. Using a 9,280 cDNA clone array, we have compared steady-state RNA from the adaxial side from which embryos develop and from the abaxial callus at five time points over the course of the 4 weeks necessary for the development of globular embryos. In a second set of experiments, we have profiled the expression of each clone in the adaxial side during the same period. A total of 495 genes differentially expressed in at least one of these experiments were grouped according to the similarity of their expression profiles using a nonhierarchical clustering algorithm. Our results indicate that the appearance of somatic embryos is preceded by dedifferentiation of the cotyledon during the first 2 weeks on auxin. Changes in mRNA abundance of genes characteristic of oxidative stress and genes indicative of cell division in the adaxial side of the cotyledons suggest that the arrangement of the new cells into organized structures might depend on a genetically controlled balance between cell proliferation and cell death. Our data also suggest that the formation of somatic globular embryos is accompanied by the transcription of storage proteins and the synthesis of gibberellic acid. PMID- 12746520 TI - Enhancement of plant-microbe interactions using a rhizosphere metabolomics-driven approach and its application in the removal of polychlorinated biphenyls. AB - Persistent organic pollutants, such as polychlorinated biphenyls (PCBs), are a global problem. We demonstrate enhanced depletion of PCBs using root-associated microbes, which can use plant secondary metabolites, such as phenylpropanoids. Using a "rhizosphere metabolomics" approach, we show that phenylpropanoids constitute 84% of the secondary metabolites exuded from Arabidopsis roots. Phenylpropanoid-utilizing microbes are more competitive and are able to grow at least 100-fold better than their auxotrophic mutants on roots of plants that are able to synthesize or overproduce phenylpropanoids, such as flavonoids. Better colonization of the phenylpropanoid-utilizing strain in a gnotobiotic system on the roots of flavonoid-producing plants leads to almost 90% removal of PCBs in a 28-d period. Our work complements previous approaches to engineer soil microbial populations based on opines produced by transgenic plants and used by microbes carrying opine metabolism genes. The current approach based on plant natural products can be applied to contaminated soils with pre-existing vegetation. This strategy is also likely to be applicable to improving the competitive abilities of biocontrol and biofertilization strains. PMID- 12746521 TI - The cotton kinesin-like calmodulin-binding protein associates with cortical microtubules in cotton fibers. AB - Microtubules in interphase plant cells form a cortical array, which is critical for plant cell morphogenesis. Genetic studies imply that the minus end-directed microtubule motor kinesin-like calmodulin-binding protein (KCBP) plays a role in trichome morphogenesis in Arabidopsis. However, it was not clear whether this motor interacted with interphase microtubules. In cotton (Gossypium hirsutum) fibers, cortical microtubules undergo dramatic reorganization during fiber development. In this study, cDNA clones of the cotton KCBP homolog GhKCBP were isolated from a cotton fiber-specific cDNA library. During cotton fiber development from 10 to 21 DPA, the GhKCBP protein level gradually decreases. By immunofluorescence, GhKCBP was detected as puncta along cortical microtubules in fiber cells of different developmental stages. Thus our results provide evidence that GhKCBP plays a role in interphase cell growth likely by interacting with cortical microtubules. In contrast to fibers, in dividing cells of cotton, GhKCBP localized to the nucleus, the microtubule preprophase band, mitotic spindle, and the phragmoplast. Therefore KCBP likely exerts multiple roles in cell division and cell growth in flowering plants. PMID- 12746522 TI - A novel family in Medicago truncatula consisting of more than 300 nodule-specific genes coding for small, secreted polypeptides with conserved cysteine motifs. AB - Transcriptome analysis of Medicago truncatula nodules has led to the discovery of a gene family named NCR (nodule-specific cysteine rich) with more than 300 members. The encoded polypeptides were short (60-90 amino acids), carried a conserved signal peptide, and, except for a conserved cysteine motif, displayed otherwise extensive sequence divergence. Family members were found in pea (Pisum sativum), broad bean (Vicia faba), white clover (Trifolium repens), and Galega orientalis but not in other plants, including other legumes, suggesting that the family might be specific for galegoid legumes forming indeterminate nodules. Gene expression of all family members was restricted to nodules except for two, also expressed in mycorrhizal roots. NCR genes exhibited distinct temporal and spatial expression patterns in nodules and, thus, were coupled to different stages of development. The signal peptide targeted the polypeptides in the secretory pathway, as shown by green fluorescent protein fusions expressed in onion (Allium cepa) epidermal cells. Coregulation of certain NCR genes with genes coding for a potentially secreted calmodulin-like protein and for a signal peptide peptidase suggests a concerted action in nodule development. Potential functions of the NCR polypeptides in cell-to-cell signaling and creation of a defense system are discussed. PMID- 12746523 TI - Chimeric RNA/DNA oligonucleotide-based site-specific modification of the tobacco acetolactate syntase gene. AB - Single amino acid substitutions at either of two crucial positions in acetolactate synthase (ALS) result in a chlorsulfuron-insensitive form of this enzyme and, as a consequence, a herbicide-resistant phenotype. Here, we describe the successful in vivo targeting of endogenous tobacco (Nicotiana tabacum) ALS genes using chimeric RNA/DNA and all-DNA oligonucleotides at two different locations. Similar number of conversion events with two different chimeras indicates the absence of restricting influence of genomic target sequence on the gene repair in tobacco. Chlorsulfuron-resistant plants were regenerated from calli after mesophyll protoplast electroporation or leaf tissue particle bombardment with these specifically constructed chimeras. Sequence analysis and enzyme assays proved the resulting alterations to ALS at both DNA and protein levels. Furthermore, foliar application of chlorsulfuron confirmed the development of resistant phenotypes. Lines with proline-196-alanine, threonine, glutamine, or serine substitutions or with tryptophan-573-leucine substitutions were highly resistant at both cellular and whole plant levels, whereas lines with proline-196-leucine substitutions were less resistant. The stability of these modifications was demonstrated by the continuous growth of calli on chlorsulfuron containing medium and by the transmission of herbicide resistance to progeny in a Mendelian manner. Ability of haploid state to promote chimera-mediated conversions is discussed. PMID- 12746524 TI - Ethylene insensitivity modulates ozone-induced cell death in birch. AB - We have used genotypic variation in birch (Betula pendula Roth) to investigate the roles of ozone (O(3))-induced ethylene (ET), jasmonic acid, and salicylic acid in the regulation of tissue tolerance to O(3). Of these hormones, ET evolution correlated best with O(3)-induced cell death. Disruption of ET perception by transformation of birch with the dominant negative mutant allele etr1-1 of the Arabidopsis ET receptor gene ETR1 or blocking of ET perception with 1-methylcyclopropene reduced but did not completely prevent the O(3)-induced cell death, when inhibition of ET biosynthesis with aminooxyacetic acid completely abolished O(3) lesion formation. This suggests the presence of an ET-signaling independent but ET biosynthesis-dependent component in the ET-mediated stimulation of cell death in O(3)-exposed birch. Functional ET signaling was required for the O(3) induction of the gene encoding beta-cyanoalanine synthase, which catalyzes detoxification of the cyanide formed during ET biosynthesis. The results suggest that functional ET signaling is required to protect birch from the O(3)-induced cell death and that a decrease in ET sensitivity together with a simultaneous, high ET biosynthesis can potentially cause cell death through a deficient detoxification of cyanide. PMID- 12746525 TI - Differential effects of elevated ozone on two hybrid aspen genotypes predisposed to chronic ozone fumigation. Role of ethylene and salicylic acid. AB - The role of ethylene (ET) signaling in the responses of two hybrid aspen (Populus tremula L. x P. tremuloides Michx.) clones to chronic ozone (O(3); 75 nL L(-1)) was investigated. The hormonal responses differed between the clones; the O(3) sensitive clone 51 had higher ET evolution than the tolerant clone 200 during the exposure, whereas the free salicylic acid concentration in clone 200 was higher than in clone 51. The cellular redox status, measured as glutathione redox balance, did not differ between the clones suggesting that the O(3) lesions were not a result of deficient antioxidative capacity. The buildup of salicylic acid during chronic O(3) exposure might have prevented the up-regulation of ET biosynthesis in clone 200. Blocking of ET perception with 1-methylcyclopropene protected both clones from the decrease in net photosynthesis during chronic exposure to O(3). After a pretreatment with low O(3) for 9 d, an acute 1.5-fold O(3) elevation caused necrosis in the O(3)-sensitive clone 51, which increased substantially when ET perception was blocked. The results suggest that in hybrid aspen, ET signaling had a dual role depending on the severity of the stress. ET accelerated leaf senescence under low O(3), but under acute O(3) elevation, ET signaling seemed to be required for protection from necrotic cell death. PMID- 12746526 TI - Apoplasmic barriers and oxygen transport properties of hypodermal cell walls in roots from four amazonian tree species. AB - The formation of suberized and lignified barriers in the exodermis is suggested to be part of a suite of adaptations to flooded or waterlogged conditions, adjusting transport of solutes and gases in and out of roots. In this study, the composition of apoplasmic barriers in hypodermal cell walls and oxygen profiles in roots and the surrounding medium of four Amazon tree species that are subjected to long-term flooding at their habitat was analyzed. In hypodermal cell walls of the deciduous tree Crateva benthami, suberization is very weak and dominated by monoacids, 2-hydroxy acids, and omega-hydroxycarboxylic acids. This species does not show any morphological adaptations to flooding and overcomes the aquatic period in a dormant state. Hypodermal cells of Tabernaemontana juruana, a tree which is able to maintain its leaf system during the aquatic phase, are characterized by extensively suberized walls, incrusted mainly by the unsaturated C(18) omega-hydroxycarboxylic acid and the alpha,omega-dicarboxylic acid analogon, known as typical suberin markers. Two other evergreen species, Laetia corymbulosa and Salix martiana, contained 3- to 4-fold less aliphatic suberin in the exodermis, but more than 85% of the aromatic moiety of suberin are composed of para-hydroxybenzoic acid, suggesting a function of suberin in pathogen defense. No major differences in the lignin content among the species were observed. Determination of oxygen distribution in the roots and rhizosphere of the four species revealed that radial loss of oxygen can be effectively restricted by the formation of suberized barriers but not by lignification of exodermal cell walls. PMID- 12746527 TI - Inorganic carbon limitation and light control the expression of transcripts related to the CO2-concentrating mechanism in the cyanobacterium Synechocystis sp. strain PCC6803. AB - The cyanobacterium Synechocystis sp. strain PCC6803 possesses three modes of inorganic carbon (Ci) uptake that are inducible under Ci stress and that dramatically enhance the efficiency of the CO(2)-concentrating mechanism (CCM). The effects of Ci limitation on the mRNA transcript abundance of these inducible uptake systems and on the physiological expression of the CCM were investigated in detail in this cyanobacterium. Transcript abundance was assessed with semiquantitative and real-time reverse transcriptase-polymerase chain reaction techniques. Cells aerated with CO(2)-free air for 30 min in the light, but not in the dark, depleted the total [Ci] to near zero levels. Under these conditions, the full physiological expression of the CCM was apparent within 2 h. Transcripts for the three inducible Ci uptake systems, ndhF3, sbtA, and cmpA, showed near maximal abundance at 15 min under Ci limitation. The transcriptional regulators, cmpR and ndhR, were more moderately expressed, whereas the rbcLXS and ccmK-N operons and ndhF4/ndhD4/chpX and ccaA genes were insensitive to the low-Ci treatment. The combined requirement of low Ci and light for the expression of several CCM-related transcripts was examined using real-time reverse transcriptase-polymerase chain reaction. CmpA, ndhF3, and sbtA were strongly expressed in the light, but not in the dark, under low-Ci conditions. We could find no evidence for induction of these or other CCM-related genes by a high light treatment under high-CO(2) conditions. This provided evidence that high light stress alone could not trigger the expression of CCM-related transcripts in Synechocystis sp. PCC6803. Potential signals triggering induction of the high affinity state of the CCM are discussed. PMID- 12746528 TI - Towards an analysis of the rice mitochondrial proteome. AB - Purified rice (Oryza sativa) mitochondrial proteins have been arrayed by isoelectric focusing/polyacrylamide gel electrophoresis (PAGE), by blue-native (BN) PAGE, and by reverse-phase high-performance liquid chromatography (LC) separation (LC-mass spectrometry [MS]). From these protein arrays, we have identified a range of rice mitochondrial proteins, including hydrophilic/hydrophobic proteins (grand average of hydropathicity = -1.27 to +0.84), highly basic and acid proteins (isoelectric point = 4.0-12.5), and proteins over a large molecular mass range (6.7-252 kD), using proteomic approaches. BN PAGE provided a detailed picture of electron transport chain protein complexes. A total of 232 protein spots from isoelectric focusing/PAGE and BN PAGE separations were excised, trypsin digested, and analyzed by tandem MS (MS/MS). Using this dataset, 149 of the protein spots (the products of 91 nonredundant genes) were identified by searching translated rice open reading frames from genomic sequence and six-frame translated rice expressed sequence tags. Sequence comparison allowed us to assign functions to a subset of 85 proteins, including many of the major function categories expected for this organelle. A further six spots were matched to rice sequences for which no specific function has yet been determined. Complete digestion of mitochondrial proteins with trypsin yielded a peptide mixture that was analyzed directly by reverse-phase LC via organic solvent elution from a C-18 column (LC-MS). These data yielded 170 MS/MS spectra that matched 72 sequence entries from open reading frame and expressed sequence tag databases. Forty-five of these were obtained using LC-MS alone, whereas 28 proteins were identified by both LC-MS and gel based separations. In total, 136 nonredundant rice proteins were identified, including a new set of 23 proteins of unknown function located in plant mitochondria. We also report the first direct identification, to our knowledge, of PPR (pentatricopeptide repeat) proteins in the plant mitochondrial proteome. This dataset provides the first extensive picture, to our knowledge, of mitochondrial functions in a model monocot plant. PMID- 12746529 TI - Isolation and characterization of the neutral leucine aminopeptidase (LapN) of tomato. AB - Tomatoes (Lycopersicon esculentum) express two forms of leucine aminopeptidase (LAP-A and LAP-N) and two LAP-like proteins. The relatedness of LAP-N and LAP-A was determined using affinity-purified antibodies to four LAP-A protein domains. Antibodies to epitopes in the most N-terminal region were able to discriminate between LAP-A and LAP-N, whereas antibodies recognizing central and COOH-terminal regions recognized both LAP polypeptides. Two-dimensional immunoblots showed that LAP-N and the LAP-like proteins were detected in all vegetative (leaves, stems, roots, and cotyledons) and reproductive (pistils, sepals, petals, stamens, and floral buds) organs examined, whereas LAP-A exhibited a distinct expression program. LapN was a single-copy gene encoding a rare-class transcript. A full length LapN cDNA clone was isolated, and the deduced sequence had 77% peptide sequence identity with the wound-induced LAP-A. Comparison of LAP-N with other plant LAPs identified 28 signature residues that classified LAP proteins as LAP-N or LAP-A like. Overexpression of a His(6)-LAP-N fusion protein in Escherichia coli demonstrated distinct differences in His(6)-LAP-N and His(6)-LAP-A activities. Similar to LapA, the LapN RNA encoded a precursor protein with a molecular mass of 60 kD. The 5-kD presequence had features similar to plastid transit peptides, and processing of the LAP-N presequence could generate the mature 55-kD LAP-N. Unlike LapA, the LapN transcript contained a second in-frame ATG, and utilization of this potential initiation codon would yield a 55-kD LAP-N protein. The localization of LAP-N could be controlled by the balance of translational initiation site utilization and LAP-N preprotein processing. PMID- 12746530 TI - The activity of the chloroplastic Ndh complex is regulated by phosphorylation of the NDH-F subunit. AB - Hydrogen peroxide (H(2)O(2)) induces increases, to different degrees, in transcripts, protein levels, and activity of the Ndh complex (EC 1.6.5.3). In the present work, we have compared the effects of relatively excess light, H(2)O(2), dimethylthiourea (a scavenger of H(2)O(2)), and/or EGTA (a Ca(2+) chelator) on the activity and protein levels of the Ndh complex of barley (Hordeum vulgare cv Hassan) leaf segments. The results show the involvement of H(2)O(2) in the modulation of both the protein level and activity of the Ndh complex and the participation of Ca(2+) mainly in the activity regulation of pre-existing protein. Changes in Ndh complex activity could not be explained only by changes in Ndh protein levels, suggesting posttranslational modifications. Hence, we investigate the possible phosphorylation of the Ndh complex both in thylakoids and in the immunopurified Ndh complex using monoclonal phosphoamino acid antibodies. We demonstrate that the Ndh complex is phosphorylated in vivo at threonine residue(s) of the NDH-F polypeptide and that the level of phosphorylation is closely correlated with the Ndh complex activity. The emerging picture is that full activity of the Ndh complex is reached by phosphorylation of its NDH-F subunit in a H(2)O(2)- and Ca(2+)-mediated action. PMID- 12746532 TI - Salicylic acid alleviates the cadmium toxicity in barley seedlings. AB - Salicylic acid (SA) plays a key role in plant disease resistance and hypersensitive cell death but is also implicated in hardening responses to abiotic stressors. Cadmium (Cd) exposure increased the free SA contents of barley (Hordeum vulgare) roots by a factor of about 2. Cultivation of dry barley caryopses presoaked in SA-containing solution for only 6 h or single transient addition of SA at a 0.5 mM concentration to the hydroponics solution partially protected the seedlings from Cd toxicity during the following growth period. Both SA treatments had little effect on growth in the absence of Cd, but increased root and shoot length and fresh and dry weight and inhibited lipid peroxidation in roots, as indicated by malondialdehyde contents, in the presence of Cd. To test whether this protection was due to up-regulation of antioxidant enzymes, activities and transcript levels of the H(2)O(2)-metabolizing enzymes such as catalase and ascorbate peroxidase were measured in control and SA-treated seedlings in the presence or absence of 25 microM Cd. Cd stress increased the activity of these enzymes by variable extent. SA treatments strongly or completely suppressed the Cd-induced up-regulation of the antioxidant enzyme activities. Slices from leaves treated with SA for 24 h also showed an increased level of tolerance toward high Cd concentrations as indicated by chlorophyll a fluorescence parameters. The results support the conclusion that SA alleviates Cd toxicity not at the level of antioxidant defense but by affecting other mechanisms of Cd detoxification. PMID- 12746531 TI - Quantitative trait loci and comparative genomics of cereal cell wall composition. AB - Quantitative trait loci (QTLs) affecting sugar composition of the cell walls of maize (Zea mays) pericarp were mapped as an approach to the identification of genes involved in cereal wall biosynthesis. Mapping was performed using the IBM (B73 x Mo17) recombinant inbred line population. There were statistically significant differences between B73 and Mo17 in content of xylose (Xyl), arabinose (Ara), galactose (Gal), and glucose. Thirteen QTLs were found, affecting the content of Xyl (two QTLs), Ara (two QTLs), Gal (five QTLs), Glc (two QTLs), Ara + Gal (one QTL), and Xyl + Glc (one QTL). The chromosomal regions corresponding to two of these, affecting Ara + Gal and Ara on maize chromosome 3, could be aligned with a syntenic region on rice (Oryza sativa) chromosome 1, which has been completely sequenced and annotated. The contiguous P1-derived artificial chromosome rice clones covering the QTLs were predicted to encode 117 and 125 proteins, respectively. Two of these genes encode putative glycosyltransferases, displaying similarity to carbohydrate-active enzyme database family GT4 (galactosyltransferases) or to family GT64 (C-terminal domain of animal heparan synthases). The results illustrate the potential of using natural variation, emerging genomic resources, and homeology within the Poaceae to identify candidate genes involved in the essential process of cell wall biosynthesis. PMID- 12746533 TI - Plant movement. Submergence-induced petiole elongation in Rumex palustris depends on hyponastic growth. AB - The submergence-tolerant species Rumex palustris (Sm.) responds to complete submergence by an increase in petiole angle with the horizontal. This hyponastic growth, in combination with stimulated elongation of the petiole, can bring the leaf tips above the water surface, thus restoring gas exchange and enabling survival. Using a computerized digital camera set-up the kinetics of this hyponastic petiole movement and stimulated petiole elongation were studied. The hyponastic growth is a relatively rapid process that starts after a lag phase of 1.5 to 3 h and is completed after 6 to 7 h. The kinetics of hyponastic growth depend on the initial angle of the petiole at the time of submergence, a factor showing considerable seasonal variation. For example, lower petiole angles at the time of submergence result in a shorter lag phase for hyponastic growth. This dependency of the hyponastic growth kinetics can be mimicked by experimentally manipulating the petiole angle at the time of submergence. Stimulated petiole elongation in response to complete submergence also shows kinetics that are dependent on the petiole angle at the time of submergence, with lower initial petiole angles resulting in a longer lag phase for petiole elongation. Angle manipulation experiments show that stimulated petiole elongation can only start when the petiole has reached an angle of 40 degrees to 50 degrees. The petiole can reach this "critical angle" for stimulated petiole elongation by the process of hyponastic growth. This research shows a functional dependency of one response to submergence in R. palustris (stimulated petiole elongation) on another response (hyponastic petiole growth), because petiole elongation can only contribute to the leaf reaching the water surface when the petiole has a more or less upright position. PMID- 12746534 TI - Generation and analysis of an artificial gene dosage series in tomato to study the mechanisms by which the cloned quantitative trait locus fw2.2 controls fruit size. AB - It has been proposed that fw2.2 encodes a negative fruit-growth regulator that underlies natural fruit-size variation in tomato (Lycopersicon spp.) via heterochronic allelic variation of fw2.2 expression, rather than by variation in the structural protein itself. To further test the negative regulator and the transcriptional control hypotheses, a gene dosage series was constructed, which produced a wider range of fw2.2 transcript accumulation than can be found in natural tomato populations. Fruit developmental analyses revealed that fw2.2 transcript levels were highly correlated (negatively) with fruit mass, supporting the negative regulator and transcriptional regulation hypotheses. Further, the effect of fw2.2 on fruit mass was mediated by repressing three- and two dimensional cell division in placental and pericarp tissues, respectively. Finally, fw2.2 had little effect on fertility and seed size/number, indicating that fruit size effects of fw2.2 are due largely to expression in the maternal tissues of developing fruit and not mediated through fertility or seed-setting related processes. PMID- 12746535 TI - Chloroplast membrane photostability in chlP transgenic tobacco plants deficient in tocopherols. AB - The phototolerance of three chlP transgenic tobacco (Nicotiana tabacum) lines, affected in geranylgeranyl reductase and, hence, deficient in tocopherols (vitamin E), was estimated by in vivo luminescence and fluorescence measurements and was compared with that of the wild type (WT). Exposure of leaf discs to high light (1 mmol photon m(-2) s(-1)) and low temperature (10 degrees C) led to a rapid inhibition of photosystem II (PSII) photochemistry that showed little dependence on the tocopherol level. PSII photo-inhibition was followed by lipid peroxidation with a time delay of about 4 h, and this phenomenon was exacerbated in the tocopherol-deficient leaves. A linear correlation was observed in these short-term experiments between resistance to photooxidation and tocopherol content. When whole plants were exposed to the same treatment, PSII was severely photo-inhibited in mature leaves of all genotypes. Lipid peroxidation was also observed in all plants, but it occurred much more rapidly in tocopherol-deficient transgenic plants relative to WT plants. The time at which extensive lipid peroxidation occurred was correlated with the tocopherol content of the leaves. The present results show that tocopherols protect thylakoid membranes against photodestruction through lipid peroxidation. However, tocopherol deficiency was compensated in young, developing leaves that were able to photo-acclimate in the long term and did not suffer from photooxidative damage. Soluble antioxidants (glutathione and ascorbate) did not accumulate in photo-acclimated chlP transgenic leaves relative to WT leaves. In contrast, a selective accumulation of xanthophyll cycle pigments was observed in young transgenic leaves, and this could represent a compensatory mechanism for tocopherol deficiency. PMID- 12746536 TI - Nod factor and elicitors activate different phospholipid signaling pathways in suspension-cultured alfalfa cells. AB - Lipo-chitooligosaccharides (Nod factors) are produced by symbiotic Rhizobium sp. bacteria to elicit Nod responses on their legume hosts. One of the earliest responses is the formation of phosphatidic acid (PA), a novel second messenger in plant cells. Remarkably, pathogens have also been reported to trigger the formation of PA in nonlegume plants. To investigate how host plants can distinguish between symbionts and pathogens, the effects of Nod factor and elicitors (chitotetraose and xylanase) on the formation of PA were investigated in suspension-cultured alfalfa (Medicago sativa) cells. Theoretically, PA can be synthesized via two signaling pathways, i.e. via phospholipase D (PLD) and via phospholipase C in combination with diacylglycerol (DAG) kinase. Therefore, a strategy involving differential radiolabeling with [(32)P]orthophosphate was used to determine the contribution of each pathway to PA formation. In support, PLD activity was specifically measured by using the ability of the enzyme to transfer the phosphatidyl group of its substrate to a primary alcohol. In practice, Nod factor, chitotetraose, and xylanase induced the formation of PA and its phosphorylated product DAG pyrophosphate within 2 min of treatment. However, whereas phospholipase C and DAG kinase were activated during treatment with all three different compounds, PLD was only activated by Nod factor. No evidence was obtained for the activation of phospholipase A(2). PMID- 12746537 TI - Identification of novel mitochondrial protein components of Chlamydomonas reinhardtii. A proteomic approach. AB - Pure mitochondria of the photosynthetic alga Chlamydomonas reinhardtii were analyzed using blue native-polyacrylamide gel electrophoresis (BN-PAGE). The major oxidative phosphorylation complexes were resolved: F(1)F(0)-ATP synthase, NADH-ubiquinone oxidoreductase, ubiquinol-cytochrome c reductase, and cytochrome c oxidase. The oligomeric states of these complexes were determined. The F(1)F(0) ATP synthase runs exclusively as a dimer, in contrast to the C. reinhardtii chloroplast enzyme, which is present as a monomer and subcomplexes. The sequence of a 60-kD protein, associated with the mitochondrial ATP synthase and with no known counterpart in any other organism, is reported. This protein may be related to the strong dimeric character of the algal F(1)F(0)-ATP synthase. The oxidative phosphorylation complexes resolved by BN-PAGE were separated into their subunits by second dimension sodium dodecyl sulfate-PAGE. A number of polypeptides were identified mainly on the basis of their N-terminal sequence. Core I and II subunits of complex III were characterized, and their proteolytic activities were predicted. Also, the heterodimeric nature of COXIIA and COXIIB subunits in cytochrome c oxidase was demonstrated. Other mitochondrial proteins like the chaperone HSP60, the alternative oxidase, the aconitase, and the ADP/ATP carrier were identified. BN-PAGE was also used to approach the analysis of the major chloroplast protein complexes of C. reinhardtii. PMID- 12746538 TI - Solubilization of an arabinan arabinosyltransferase activity from mung bean hypocotyls. AB - The biosynthesis of polysaccharides destined for the plant cell wall and the subsequent assembly of the cell wall are poorly understood processes that are currently the focus of much research. Arabinan, a component of the pectic polysaccharide rhamnogalacturonan I, is composed of arabinosyl residues connected via various glycosidic linkages, and therefore, the biosynthesis of arabinan is likely to involve more than one arabinosyltransferase. We have studied the transfer of [(14)C]arabinose (Ara) from UDP-L-arabinopyranose onto polysaccharides using microsomal membranes isolated from mung bean (Vigna radiata) hypocotyls. [(14)C]arabinosyl and [(14)C]xylosyl residues were incorporated into endogenous products due to the presence of UDP-Xyl-4-epimerase activity. Enzymatic digestion of endogenous products with endo-arabinanase released very little radiolabeled sugars, whereas digestion with arabinofuranosidase released some [(14)C]Ara. Microsomal membranes solubilized with the detergent octyl glucoside were able to add a single [(14)C]Ara residue onto (1-->5)-linked alpha-L-arabino-oligosaccharide acceptors. The reaction had a pH optimum of 6.5 and a requirement for manganese ions. However, enzymatic digestion of the radiolabeled oligosaccharides with endo-arabinanase and arabinofuranosidases could not fully release the radiolabeled Ara residue, indicating that the [(14)C]Ara residue was not a (1-->2)-, (1-->3)-, or (1-->5) linked alpha-L-arabinofuranosyl residue. Rather, mild acid treatment of the product suggested that the radiolabeled Ara residue was in a pyranose conformation, and this result was confirmed by thin-layer chromatography of radiolabeled partially methylated sugars. Using microsomal membranes separated on a discontinuous sucrose gradient, the arabinosyltransferase activity appears to be mainly localized to Golgi membranes. PMID- 12746539 TI - The expression of the t-SNARE AtSNAP33 is induced by pathogens and mechanical stimulation. AB - The fusion of vesicles in the secretory pathway involves the interaction of t soluble N-ethylmaleimide-sensitive factor attachment protein receptors (t-SNAREs) on the target membrane and v-SNAREs on the vesicle membrane. AtSNAP33 is an Arabidopsis homolog of the neuronal t-SNARE SNAP-25 involved in exocytosis and is localized at the cell plate and at the plasma membrane. In this paper, the expression of AtSNAP33 was analyzed after different biotic and abiotic stresses. The expression of AtSNAP33 increased after inoculation with the pathogens Plectosporium tabacinum and virulent and avirulent forms of Peronospora parasitica and Pseudomonas syringae pv tomato. The expression of PR1 transcripts encoding the secreted pathogenesis-related protein 1 also increased after inoculation with these pathogens and the expression of AtSNAP33 preceded or occurred at the same time as the expression of PR1. AtSNAP33 was also expressed in npr1 plants that do not express PR1 after pathogen inoculation as well as in cpr1 plants that overexpress PR1 in the absence of a pathogen. The level of AtSNAP33 decreased slightly in leaves inoculated with P. parasitica in the NahG plants, and eds5 and sid2 mutants that are unable to accumulate salicylic acid (SA) after pathogen inoculation, indicating a partial dependence on SA. AtSNAP33 was also expressed in systemic noninoculated leaves of plants inoculated with P. syringae. In contrast to the situation in infected leaves, the expression of AtSNAP33 in systemic leaves was fully SA dependent. Thus, the expression of AtSNAP33 after pathogen attack is regulated by SA-dependent and SA-independent pathways. Mechanical stimulation also led to an increase of AtSNAP33 transcripts. PMID- 12746540 TI - Role of the reversible xanthophyll cycle in the photosystem II damage and repair cycle in Dunaliella salina. AB - The Dunaliella salina photosynthetic apparatus organization and function was investigated in wild type (WT) and a mutant (zea1) lacking all beta,beta epoxycarotenoids derived from zeaxanthin (Z). The zea1 mutant lacked antheraxanthin, violaxanthin, and neoxanthin from its thylakoid membranes but constitutively accumulated Z instead. It also lacked the so-called xanthophyll cycle, which, upon irradiance stress, reversibly converts violaxanthin to Z via a de-epoxidation reaction. Despite the pronounced difference observed in the composition of beta,beta-epoxycarotenoids between WT and zea1, no discernible difference could be observed between the two strains in terms of growth, photosynthesis, organization of the photosynthetic apparatus, photo-acclimation, sensitivity to photodamage, or recovery from photo-inhibition. WT and zea1 were probed for the above parameters over a broad range of growth irradiance and upon light shift experiments (low light to high light shift and vice versa). A constitutive accumulation of Z in the zea1 strain did not affect the acclimation of the photosynthetic apparatus to irradiance, as evidenced by indistinguishable irradiance-dependent adjustments in the chlorophyll antenna size and photosystem content of WT and zea1 strain. In addition, a constitutive accumulation of Z in the zea1 strain did not affect rates of photodamage or the recovery of the photosynthetic apparatus from photo-inhibition. However, Z in the WT accumulated in parallel with the accumulation of photodamaged PSII centers in the chloroplast thylakoids and decayed in tandem with a chloroplast recovery from photo inhibition. These results suggest a role for Z in the protection of photodamaged and disassembled PSII reaction centers, apparently needed while PSII is in the process of degradation and replacement of the D1/32-kD reaction center protein. PMID- 12746541 TI - Growth ring formation in the starch granules of potato tubers. AB - Starch granules from higher plants contain alternating zones of semicrystalline and amorphous material known as growth rings. The regulation of growth ring formation is not understood. We provide several independent lines of evidence that growth ring formation in the starch granules of potato (Solanum tuberosum) tubers is not under diurnal control. Ring formation is not abolished by growth in constant conditions, and ring periodicity and appearance are relatively unaffected by a change from a 24-h to a 40-h photoperiod, and by alterations in substrate supply to the tuber that are known to affect the diurnal pattern of tuber starch synthesis. Some, but not all, of the features of ring formation are consistent with the involvement of a circadian rhythm. Such a rhythm might operate by changing the relative activities of starch-synthesizing enzymes: Growth ring formation is disrupted in tubers with reduced activity of a major isoform of starch synthase. We suggest that physical as well as biological mechanisms may contribute to the control of ring formation, and that a complex interplay of several factors may by involved. PMID- 12746542 TI - Molecular cloning and functional characterization of three distinct N methyltransferases involved in the caffeine biosynthetic pathway in coffee plants. AB - Caffeine is synthesized from xanthosine through N-methylation and ribose removal steps. In the present study, three types of cDNAs encoding N-methyltransferases were isolated from immature fruits of coffee (Coffea arabica) plants, and designated as CaXMT1, CaMXMT2, and CaDXMT1, respectively. The bacterially expressed encoded proteins were characterized for their catalytic properties. CaXMT1 catalyzed formation of 7-methylxanthosine from xanthosine with a K(m) value of 78 microM, CaMXMT2 catalyzed formation of 3,7-dimethylxanthine (theobromine) from 7-methylxanthine with a K(m) of 251 microM, and CaDXMT1 catalyzed formation of 1,3,7-trimethylxanthine (caffeine) from 3,7 dimethylxanthine with a K(m) of 1,222 microM. The crude extract of Escherichia coli was found to catalyze removal of the ribose moiety from 7-methylxanthosine, leading to the production of 7-methylxanthine. As a consequence, when all three recombinant proteins and E. coli extract were combined, xanthosine was successfully converted into caffeine in vitro. Transcripts for CaDXMT1 were predominantly found to accumulate in immature fruits, whereas those for CaXMT1 and CaMXMT2 were more broadly detected in sites encompassing the leaves, floral buds, and immature fruits. These results suggest that the presently identified three N-methyltransferases participate in caffeine biosynthesis in coffee plants and substantiate the proposed caffeine biosynthetic pathway: xanthosine --> 7 methylxanthosine --> 7-methylxanthine --> theobromine --> caffeine. PMID- 12746543 TI - Involvement of hydrogen peroxide and nitric oxide in expression of the ipomoelin gene from sweet potato. AB - The IPO (ipomoelin) gene was isolated from sweet potato (Ipomoea batatas cv Tainung 57) and used as a molecular probe to investigate its regulation by hydrogen peroxide (H(2)O(2)) and nitric oxide (NO) after sweet potato was wounded. The expression of the IPO gene was stimulated by H(2)O(2) whether or not the plant was wounded, but its expression after wounding was totally suppressed by the presence of diphenylene iodonium, an inhibitor of NADPH oxidase, both in the local and systemic leaves of sweet potato. These results imply that a signal transduction resulting from the mechanical wounding of sweet potato may involve NADPH oxidase, which produces endogenous H(2)O(2) to stimulate the expression of the IPO gene. The production of H(2)O(2) was also required for methyl jasmonate to stimulate the IPO gene expression. On the contrary, NO delayed the expression of the IPO gene, whereas N(G)-monomethyl-L-arginine monoacetate, an inhibitor of NO synthase, enhanced the expression of the IPO gene after the plant was wounded. This study also demonstrates that the production of H(2)O(2) stained with 3,3' diaminobenzidine hydrochloride could be stimulated by wounding but was suppressed in the presence of NO. Meanwhile, the generation of NO was visualized by confocal scanning microscope in the presence of 4,5-diaminofluorescein diacetate after sweet potato was wounded. In conclusion, when sweet potato was wounded, both H(2)O(2) and NO were produced to modulate the plant's defense system. Together, H(2)O(2) and NO regulate the expression of the IPO gene, and their interaction might further stimulate plants to protect themselves from invasions by pathogens and herbivores. PMID- 12746546 TI - Iron regulatory proteins as NO signal transducers. AB - The iron regulatory proteins (IRPs) are an example of different proteins regulating the same metabolic process, iron uptake and metabolism. IRP1 is an iron-sulfur cluster-containing protein that can be converted from a cytosolic aconitase to an RNA binding posttranscriptional regulator in response to nitric oxide (NO). IRP2 lacks aconitase activity and its expression is decreased by NO signaling. In macrophages, NO is produced in response to such inflammatory ligands as interferon-gamma, which is expressed in response to mitogenic and antigenic stimuli, and lipopolysaccharide, a marker of bacterial invasion. Until recently, research results predict that the cellular response to increased NO production should be a decrease in ferritin synthesis, due to IRP1 binding to ferritin mRNA, and an increase in transferrin receptor biosynthesis, due to IRP1 binding to the transferrin mRNA. Surprisingly, however, macrophages exhibit decreased transferrin receptor concentration in response to inflammatory ligands. Bouton and Drapier discuss the physiological role and the mechanisms that may underlie this contradictory response. PMID- 12746547 TI - Imaging zinc: old and new tools. AB - As recently as 20 years ago, all zinc in biological systems was believed to be tightly bound to proteins, and the idea of imaging zinc was considered heretical. Beginning with Maske's research with dithizonate staining of the hippocampus in the 1950s, however, zinc-sensitive dyes have indicated that, in mammalian cells, free zinc can exist in at least three separate pools. These pools include vesicular zinc sequestered in presynaptic vesicles and secretory granules, zinc released from these vesicles into the extracellular space after physiological stimulation, and transient increases in zinc in cells in the regions where extracellular release of zinc has occurred. This Perspective covers the zinc imaging tools, from dithizonate to the newest FRET-based sensors, that have galvanized biomedical science. PMID- 12746544 TI - Expression of the plastid-located glutamine synthetase of Medicago truncatula. Accumulation of the precursor in root nodules reveals an in vivo control at the level of protein import into plastids. AB - In this paper, we report the cloning and characterization of the plastid-located glutamine synthetase (GS) of Medicago truncatula Gaertn (MtGS2). A cDNA was isolated encoding a GS2 precursor polypeptide of 428 amino acids composing an N terminal transit peptide of 49 amino acids. Expression analysis, by Westerns and by northern hybridization, revealed that MtGS2 is expressed in both photosynthetic and non-photosynthetic organs. Both transcripts and proteins of MtGS2 were detected in substantial amounts in root nodules, suggesting that the enzyme might be performing some important role in this organ. Surprisingly, about 40% of the plastid GS in nodules occurred in the non-processed precursor form (preGS2). This precursor was not detected in any other organ studied and moreover was not observed in non-fixing nodules. Cellular fractionation of nodule extracts revealed that preGS2 is associated with the plastids and that it is catalytically inactive. Immunogold electron microscopy revealed a frequent coincidence of GS with the plastid envelope. Taken together, these results suggest a nodule specific accumulation of the GS2 precursor at the surface of the plastids in nitrogen-fixing nodules. These results may reflect a regulation of GS2 activity in relation to nitrogen fixation at the level of protein import into nodule plastids. PMID- 12746548 TI - Do we need zinc to think? AB - Chelatable Zn(2+), which is found in the synaptic vesicles of certain glutamatergic neurons in several regions of the forebrain, is released during neuronal activity. Zn(2+) exhibits numerous effects on ligand-gated and voltage dependent ion channels, and released Zn(2+) is therefore likely able to modulate synaptic transmission. The physiologically relevant actions of Zn(2+), however, have remained unclear. Recent research exploiting improved Zn(2+)-sensitive optical probes has suggested some intriguing effects for synaptically released Zn(2+), including heterosynaptic regulation of N-methyl-D-aspartate (NMDA) receptor function, and a novel role as a trans-synaptic second messenger that may enter postsynaptic neurons to modulate various signal transduction pathways. PMID- 12746549 TI - Coronavirus main proteinase (3CLpro) structure: basis for design of anti-SARS drugs. AB - A novel coronavirus has been identified as the causative agent of severe acute respiratory syndrome (SARS). The viral main proteinase (Mpro, also called 3CLpro), which controls the activities of the coronavirus replication complex, is an attractive target for therapy. We determined crystal structures for human coronavirus (strain 229E) Mpro and for an inhibitor complex of porcine coronavirus [transmissible gastroenteritis virus (TGEV)] Mpro, and we constructed a homology model for SARS coronavirus (SARS-CoV) Mpro. The structures reveal a remarkable degree of conservation of the substrate-binding sites, which is further supported by recombinant SARS-CoV Mpro-mediated cleavage of a TGEV Mpro substrate. Molecular modeling suggests that available rhinovirus 3Cpro inhibitors may be modified to make them useful for treating SARS. PMID- 12746550 TI - Stabilization of quaternary structure of water-soluble quinoprotein glucose dehydrogenase. AB - Water-soluble quinoprotein glucose dehydrogenase (PQQGDH-B) is a dimeric enzyme whose application for glucose sensing is the focus of much attention. We attempted to increase the thermal stability of PQQGDH-B by introducing a disulfide bond at the dimer interface. The Ser residue at position 415 was selected for substitution with Cys, as structural information revealed that its side chains face each other at the dimer interface of PQQGDH-B. PQQGDH-B with Ser415Cys showed 30-fold greater thermal stability at 55 degrees C than did the wild-type enzyme without any decrease in catalytic activity. After incubation at 70 degrees C for 10 min, Ser415Cys retained 90% of the GDH activity of the wild type enzyme. Disulfide bond formation between the mutant subunits was confirmed by analyses with sodium dodecylsulfate-polyacrylamide gel electrophoresis in the presence and absence of reductants. Our results indicate that the introduction of one Cys residue in each monomer of PQQGDH-B resulted in formation of a disulfide bond at the dimer interface and thus achieved a large increase in the thermal stability of the enzyme. PMID- 12746551 TI - Efficient mutagenesis method for producing the templates of single nucleotide polymorphisms. AB - DNA templates harboring specific single nucleotide polymorphism (SNP) sites are largely needed as positive controls in practical SNP analysis and in determination of the reliability of newly developed methods in high-throughput screening assays. Here we report a one-step method to produce SNP templates by amplifying a wild-type sequence with primers having single nucleotide mismatches at or near their 3' ends. A short amplicon harboring an EcoRI site was used to evaluate the feasibility of our strategy. Perfectly matched primers and primers with a single base mismatch occurring from the first base to the sixth base of the EcoRI site were used for primer extension. By using polymerase without a proofreading function, we kept mismatched nucleotides from occurring in extended primer products, as confirmed by EcoRI digestion and sequencing analysis. The strategy of using primers with a single mismatched base and exo- polymerase was shown to be an efficient one-step method for preparing SNP templates, either for application in the development of SNP screening assays or as positive controls in practical SNP assays. PMID- 12746552 TI - Varying the nucleic acid composition of siRNA molecules dramatically varies the duration and degree of gene silencing. AB - The utility of short interfering RNA (siRNA) as a means of gene silencing depends on several factors. These include the degree to which a gene can be silenced, the length of time for which the gene remains silenced, the degree of recovery of gene function, and the effects of the silencing process on general cell functions. We hypothesized that changing the nucleic acid composition of the siRNA constructs used for silencing would affect these parameters. With siRNA gene silencing of the glucose-6-phosphate dehydrogenase gene as a baseline, we found that siDNA molecules have an effect that is similar in duration but lesser in degree, whereas hybrid DNA:RNA molecules have an effect that is enormously greater in both duration and degree. PMID- 12746553 TI - Promoter of the rolC gene of Agrobacterium rhizogenes can be strongly regulated in glandular cell of transgenic tobacco. AB - A 577-bp promoter segment of Agrobacterium rhizogenes rolC, previously known as the phloem-specific gene expression promoter, was fused to the 5' end of a reporter gene, beta-glucuronidase (GUS), uidA. This rolC-promoter-driven expression of the GUS gene was found to be significantly strong in glandular cells in transgenic tobacco plants. Analysis of this segment of the promoter sequence revealed a myb response element. PMID- 12746554 TI - Evaluation of optimal expression cassette in retrovirus vector for beta thalassemia gene therapy. AB - Trials of retroviral vector-mediated human beta-globin gene transfer were hampered by low titers, unstable vector transmission, and low-level expression of transferred gene. With the goal of optimizing the retrovirally encoded human beta globin gene expression cassette for gene therapy of beta-thalassemia, we generated 3 series of vector constructs (a total of 12 constructs) and investigated the effects of the proximal promoter, 3' - enhancer, and derivatives from the beta-locus control region or alpha-major regulatory element on virus titer, vector transmission stability, and gene expression. The virus titers for 9 of the 12 vector constructs ranged between 2.8 x 10(4) cfu/mL and 1.0 x 10(6) cfu/mL. We found that proviral DNA was intact in most G418- resistant murine erythroleukemia (MEL) cell clones for 5 vector constructs, while obvious genetic instability was observed for 4 other vector constructs. MEL cells harboring the intact provirus were induced to differentiate, and human beta-globin gene expression was analyzed with RNase protection assay. The percentage of human beta globin transcript relative to endogenous murine alpha-globin transcript were 101.8 +/- 64.3% (n = 10), 40.1 +/- 28.7% (n = 4), 31.1 +/- 31.9% (n = 12), 52.4 +/- 11.2% (n = 12), and 53.6 +/- 8.6% (n = 12) for the 5 constructs, respectively, demonstrating the development of optimized retroviral vectors for beta-globin gene therapy with murine erythroid cell lines as a model. Unexpectedly, we also documented that the point mutation 8700(C-->T) in DNase I hypersensitive site 2 (HS2) core fragment might contribute to low-level expression of the human beta-globin gene, based on a comparison of results from transfected and transduced MEL cells and sequence analysis of proviral DNA. PMID- 12746555 TI - Clinical implication of p53 mutation in lung cancer. AB - Lung cancer development involves multiple genetic abnormalities leading to malignant transformation of the bronchial epithelial cells, followed by invasion and metastasis. One of the most common changes is mutation of the p53 tumor suppressor gene. The frequency of p53 alterations in lung cancer is highest in small cell and squamous cell carcinomas. A genetic "signature" of the type of p53 mutations has been associated with carcinogens in cigarette smoke. The majority of clinical studies suggest that lung cancers with p53 alterations carry a worse prognosis, and may be relatively more resistant to chemotherapy and radiation. An understanding of the role of p53 in human lung cancer may lead to more rational targeted approaches for treating this disease. P53 gene replacement is currently under clinical investigation but clearly more effective means of gene deliver to the tumor cells are required. Novel approaches to lung cancer therapy are needed to improve the observed poor patient survival despite current therapies. PMID- 12746557 TI - Multilocus enzyme electrophoresis: a practical guide. AB - Multilocus enzyme electrophoresis (MEE) uses the relative electrophoretic mobilities of intracellular enzymes to characterize and differentiate organisms by generating an electromorph type (ET). This article presents the chemical conditions that may be useful, a guide to the successful practice of the electrophoretic technique, and analysis of the results. PMID- 12746558 TI - A novel method for preparing single-stranded DNA for pyrosequencing. AB - Pyrosequencing technology is a powerful genotyping tool that requires the generation of single stranded DNA. Currently, two simple, solid-phase-based methods are available for this, but they require special equipment, they are not automated, and they are relatively expensive because of the need for biotinylated polymerase chain reaction primers. In this article, an enzymatic liquid-phase method for the generation of high-quality, single-stranded DNA, and its novel use for Pyrosequencing are described. The method has also been fully automated. PMID- 12746560 TI - Why is the adrenal adrenergic? AB - The adrenal gland is the body's primary source for epinephrine production and release, and the chromaffin cells that comprise the adrenal medulla possess all of the catecholamine biosynthetic machinery, including phenylethanolamine N methyltransferase (PNMT), the enzyme synthesizing epinephrine from norepinephrine. In most species, epinephrine, also known as adrenaline, is the predominant neurotransmitter/neurohormone expressed by chromaffin cells. Present knowledge about "what makes the adrenal adrenergic" is derived from studies of normal and neoplastic adrenal medullary tissue and cells, with the PNMT gene serving as a marker of adrenergic function. The preference for adrenergic expression occurs, in part, because of the juxtaposition of adrenal medulla and adrenal cortex, as the cortex provides high circulating levels of glucocorticoids to the medulla. However, although glucocorticoids and the activated glucocorticoid receptor clearly are critical elements, they are apparently not the sole components defining the adrenergic phenotype. Other factors may include several transcriptional activators of the PNMT gene: Egr-1, AP2, Sp1, and MAZ. The existence of transcription factors that silence PNMT expression in noradrenergic cells has also been postulated. Understanding the requirements for adrenergic expression may provide important insights and potential therapies for disorders in which adrenergic/catecholaminergic dysfunction leads to illnesses refractory to present treatment strategies. PMID- 12746562 TI - Hormone markers in pituitary adenomas: changes within last decade resulting from improved method. AB - The significance of polyclonal antibodies for demonstration of hormone expression in pituitary adenomas was compared with the significance of monoclonal antibodies (MAbs). Adenomas were classified by light microscopic structures in paraffin- and epon-embedded sections, by immunostaining for all pituitary hormones, and in part by electron microscopy. In the first series, 166 adenomas were studied with polyclonal antibodies during 1990 and 1991. In the second series, 572 adenomas were immunostained with MAbs during 1999 and 2000. In the first series, a very typical type-specific hormone expression was demonstrable in 63.2% of adenomas in contrast to 91.1% in the second series. Consequently, in the first series 36.7% of adenomas expressed additional or atypical hormones in >10% of adenoma cells, whereas in the second series only 8.9% contained additional hormones. We conclude that MAbs show a clearer cutoff in immunostainings of pituitary hormones and therefore should be preferred for adenoma classification. PMID- 12746559 TI - The chromogranins: their roles in secretion from neuroendocrine cells and as markers for neuroendocrine neoplasia. AB - Chromogranins are the major components of the secretory granules of most neuroendocrine cells. Within the secretory pathway, chromogranins are involved in granulogenesis, and in sorting and processing of secretory protein cargo prior to secretion. Once secreted, they have hormonal, autocrine, and paracrine activities. The chromogranin family includes chromogranins A (CgA) and B (CgB) and secretogranin II (SgII, once called chromogranin C). The related "granins" NESP55, 7B2, secretogranin III/1B 1075 (SgIII), and secretogranin IV/HISL-19 antigen (SgIV), are also sometimes included when considering the chromogranins. While it is useful to consider the granin proteins as a family with many common features, it is also necessary to examine the distinct features and properties of individual members of the granin family to understand fully their functions, employ them efficiently as tissue, serum, and urinary markers for neuroendocrine neoplasia, and develop an evolutionary-biologic perspective on their contribution to mammalian physiology. Recent advances in chromogranin research include establishing the role of CgA in granulogenesis and the role of CgB in nuclear transcription; new biologic activities for CgA-, CgB-, and SgII-derived peptides; and new marker functions for granins and their proteolytically processed products in endocrine neoplasias. PMID- 12746556 TI - Structural aspects of the metzincin clan of metalloendopeptidases. AB - Metalloendopeptidases are present across all kingdoms of living organisms; they are ubiquitous and widely involved in metabolism regulation through their ability either to extensively degrade proteins or to selectively hydrolyze specific peptide bonds. They must be subjected to exquisite spatial and temporal control to prevent this vast potential from becoming destructive. These enzymes are mostly zinc-dependent and the majority of them, named zincins, possess a short consensus sequence, HEXXH, with the two histidines acting as ligands of the catalytic zinc and the glutamate as the general base. A subclass of the zincins is characterized by a C-terminally elongated motif, HEXXHXXGXXH/D, with an additional strictly conserved glycine and a third zinc-binding histidine or aspartate. Currently, representative three-dimensional structures of six different proteinase families bearing this motif show, despite low sequence similarity, comparable overall topology. This includes a substrate-binding crevice, which subdivides the enzyme moiety into an upper and a lower subdomain. A common five-stranded beta-sheet and two alpha-helices are always found in the upper subdomain. The second of these helices encompasses the first half of the elongated consensus sequence and is therefore termed the active-site helix. Other shared characteristics are an invariant methionine-containing Met-turn beneath the catalytic metal and a further C-terminal helix in the lower subdomain. All these structural features identify the metzincin clan of metalloendopeptidases. This clan is reviewed from a structural point of view, based on the reported structures of representative members of the astacins, adamalysins, serralysins, matrixins, snapalysins, and leishmanolysins, and of inhibited forms, either by specific endogenous protein inhibitors or by zymogenic pro-domains. Moreover, newly available genomic sequences have unveiled novel putative metzincin families and new hypothetical members of existing ones. PMID- 12746563 TI - Distinction between papillary thyroid hyperplasia and papillary thyroid carcinoma by immunohistochemical staining for cytokeratin 19, galectin-3, and HBME-1. AB - The histopathology of papillary thyroid hyperplasia and papillary thyroid carcinoma is similar enough to cause a diagnostic dilemma in a few cases. Both lesions may have papillary fronds with fibrovascular cores, nuclear crowding, and nuclear anisocytosis. Formalin- fixed paraffin-embedded tissues from 30 randomly selected patients with papillary thyroid hyperplasia and an equal number from patients with papillary thyroid carcinoma were analyzed for expression of cytokeratin 19 (CK19), galectin-3, and HBME-1. Cases of papillary thyroid carcinoma had moderate to strong CK19, galectin-3, and HBME-1 reactivity although both CK19 and galectin-3 showed positive staining in a significant number of nonneoplastic thyroid cases. HBME-1 was uncommon in the nonneoplastic cases. These results indicate that HBME-1 may be useful in helping to distinguish papillary thyroid carcinoma from hyperplasia in diagnostically difficult cases. PMID- 12746561 TI - Chromogranin a processing in human pituitary adenomas and carcinomas: analysis with region-specific antibodies. AB - The expression of various chromogranin A (CgA) peptide fragments was examined with region-specific antisera in benign and malignant pituitary tumors. Analysis of the proconvertases responsible for proteolytic processing of CgA, prohormone convertase 1/3 (PC1/3), and PC2 was also performed. Adenomas were studied using tissue microarrays, and a larger tissue section of a subset of the prolactin (PRL) adenomas was used to compare to the tissue microarray analysis. Carcinomas were analyzed using larger tissue sections. There were differences in CgA proteolytic products detected between the functional (PRL, adrenocorticotropic hormone [ACTH], and growth hormone tumors and the nonfunctional (gonadotroph and null cell) tumors, with the former group expressing lower levels of many peptides. These differences were most notable in the PRL adenomas and carcinomas in which the region-specific antisera against vasostatin I and vasostatin II detected these fragments in the lowest percentage of tumors and/or had the weakest immunoreactivity. The CgA peptide fragment detected by CgA 176-195 (chromacin) antiserum was expressed by the highest percentage of most functional and nonfunctional benign and malignant pituitary tumors. ACTH carcinomas (n = 3) were more strongly immunoreactive compared to the ACTH adenomas. These results show that there is differential expression of CgA peptide fragments and PC1/3 among different types of pituitary tumors and that ACTH pituitary carcinomas have higher levels of immunoreactive CgA peptide fragments compared to ACTH adenomas. This study also shows the utility of tissue microarrays in the analysis of a large group of tumors with regionspecific antisera. PMID- 12746564 TI - Decreased expression of calcium receptor in parathyroid tissue in patients with hyperparathyroidism secondary to chronic renal failure. AB - The response of parathyroid cells to serum calcium is regulated by a calcium sensing receptor protein (CaR). In patients with chronic renal failure, hypocalcemia contributes to the parathyroid hyperplasia and increased parathyroid hormone secretion characteristic of secondary hyperparathyroidism (sHPT). However, patients with uremia also display reduced sensitivity to extracellular calcium; this seems to be owing to an alteration of the receptor mechanism. This study examined calcium receptor expression in the parathyroid tissue of patients with sHPT, using immunohistochemical techniques and comparison with normal tissue and parathyroid glands of patients with primary hyperparathyroidism. In all the glands studied, immunostaining was more intense in chief cells than in oxyphilic, transitional, and clear cells. The parathyroid glands of patients with sHPT displayed significantly reduced expression of CaR with respect to morphologically normal ones; a very similar reduction is reported in adenomas. Furthermore, in glands displaying multinodular hyperplasia, expression was less marked in nodule forming cells than in internodular areas. The decreased expression of calcium receptors in the parathyroid tissue of uremic patients was thought to be owing to the different cell populations present; these parathyroid glands contained predominantly transitional, oxyphilic, and clear cells, which normally express fewer receptors than chief cells, which are more abundant in normal glands. PMID- 12746566 TI - A comparative immunohistochemical study of spontaneous and chemically induced pheochromocytomas in B6C3F1 mice. AB - Spontaneously occurring and chemically induced pheochromocytomas are rare in mice. That the mouse pheochromocytoma is a more appropriate animal model than that of the rat for study of human medullary adrenal tumors has been suggested. The expression of phenylethanolamine-N-methyltransferase (PNMT), the enzyme responsible for production of epinephrine from norepinephrine, is common to both mouse and human pheochromocytomas. This investigation assessed the expression of the immunohistochemical markers PNMT, tyrosine hydroxylase (TH), and chromogranin A (CGA) in spontaneously occurring and chemically induced pheochromocytomas in the B6C3F1 mouse. Spontaneous tumors were derived from control animals from 10 different studies and the pheochromocytomas from treated groups from 4 different studies. All tumors were positive for maximal TH expression. A highly significant difference in PNMT expression (p < 0.01) occurred between spontaneously occurring pheochromocytomas classified as benign or "malignant" by the criteria of toxicologic pathology. Chemically induced tumors showed intermediate PNMT staining. A marked reduction in CGA expression occurred in pheochromocytomas induced by technical grade pentachlorophenol, compared to the other three chemicals and the spontaneously occurring tumors. These findings suggest that immunohistochemistry is a reliable tool in investigating the functional capabilities of pheochromocytomas in mice. PNMT expression is a tightly regulated component of the chromaffin cell phenotype and appears to be readily lost in mouse pheochromocytomas, particularly those with aggressive characteristics. PMID- 12746567 TI - An unusual case of multiple giant myelolipomas: clinical and pathogenetic implications. AB - Myelolipomas are benign tumors composed of both mature adipose and myeloid tissues. They typically present as an incidental mass in one of the adrenal glands proper. However, they can occur in ectopic adrenal tissue or, rarely, without associated adrenal tissue in various locations and can grow to weights of several kilograms. These tumors have been linked to endocrinopathies, such as Cushing disease and congenital adrenal hyperplasia, which involve overproduction of adrenocorticotropic hormone. We report a case of three giant adrenal myelolipomas arising in a persistently virilized female with congenital adrenal hyperplasia, supporting a role for hormonal stimuli in myelolipoma formation. PMID- 12746565 TI - Penetrance of inherited medullary thyroid carcinoma and genotype-phenotype correlation in a large multiple endocrine neoplasia type 2A family with C634Y RET mutation. AB - Multiple endocrine neoplasia type 2A (MEN 2A) and familial medullary thyroid carcinoma (FMTC) are characterized by development of medullary thyroid carcinoma (MTC) and caused by germline RET mutations. Patients with MEN 2A also develop pheochromocytoma and/or hyperparathyroidism (HPT). However, MEN 2A-affected individuals could display the FMTC phenotype at first clinical manifestation. To establish the correct phenotype and improve clinical management of patients affected by hereditary MTC, clinical screening, RET mutational analysis, penetrance of MTC, and genotype-phenotype correlation were performed in a large, suspected FMTC kindred of 86 individuals. Germline C634Y RET mutation was confirmed in 22 individuals, 15 of whom were thyroidectomized when high serum calcitonin levels were detected. MTC was confirmed in 12 individuals and C-cell hyperplasia in 3. HPT was detected in two patients. High penetrance of MTC at young age (79% at 30 yr of age) was found. This family was considered to be affected by FMTC for several years because MTC was the sole clinical manifestation. However, our results allowed reclassifying the family as MEN 2A, thereby improving clinical management of family members. Our findings regarding penetrance and genotype-phenotype correlation suggest that patients considered to have FMTC may in fact have MEN 2A in some kindreds. PMID- 12746568 TI - Primary malignant melanoma of adrenal gland in a 41-yr-old woman. PMID- 12746569 TI - Studies of five microelement contents in human serum, hair, and fingernails correlated with aged hypertension and coronary heart disease. AB - Using atomic absorption spectrometry (AAS), five microelements in human serum, hair, and fingernails of aged hypertension, coronary heart disease (diseased group) and aged health control (healthy group) were detected. Results of the t test are as follows: The iron, zinc, and cadmium contents and Zn/Cu (mol/mol) ratio of the diseased group were significantly higher than that of the healthy group in serum (p<0.01, p<0.05, p<0.01, and p<0.05, respectively); the chromium contents in the serum, hair, and fingernails (p<0.05, p<0.01, and p<0.05, respectively); the iron and zinc contents in the hair and fingernails (p<0.01, p<0.001, p<0.05, and p<0.01 respectively) and Zn/Cu ratio in the hair (p<0.01) of the diseased group were significantly lower than that of the healthy group. PMID- 12746570 TI - Selenium protection against mercury-induced apoptosis and growth inhibition in cultured K-562 cells. AB - Selenium and mercuric chloride (MC) interactions regarding effects on cell growth and cell death have been studied. Human K-562 cells were pretreated or simultaneously treated with either selenite (5 or 50 microM) or selenomethionine (10 or 50 microM) and with MC (35 or 50 microM). The 35-microM MC treatments resulted in a clear inhibition of cell growth with no obvious difference between mercury-treated and mercury-selenium-treated cells. Furthermore, the apoptotic frequency was similar at all observations for all selenium treatments with 35 microM MC. In the simultaneously treated selenite and 50- microM MC combinations, a selenite-dependent protection was shown both by increased cell growth and by lower apoptotic frequency at 48 and 96 h of exposure. Both treatments with selenomethionine showed protection observed as an increased cell growth at 48 and 96 h and as decreased apoptotic frequency at 96 h of exposure. PMID- 12746571 TI - Effects of chronic fluorosis on electrocardiogram in sheep. AB - This study was carried out to evaluate the effects of chronic fluorosis by means of the electrocardiograms in sheep. Ten sheep with fluorosis living around a volcanic mountain (Tendurek Mount) in East Anatolia in Turkey and 10 healthy sheep were used. Leads I, II, III, aVR, aVL, aVF, V2, V4, and V10 were recorded in the electrocardiographs of the sheep. All waves were seen in all derivations. The P-Q interval was significantly (p<0.05) prolonged and sinus bradycardia was observed in the sheep with fluorosis. As a result of this, the number of heart beats was decreased significantly (p<0.05); that is, the number of heart beats was 110 +/- 15 in the control group and 75 +/- 10 in sheep with fluorosis. PMID- 12746572 TI - Dietary zinc modifies the characteristics of endothelial dilation in normozincemic rats. AB - Because zinc attenuates endothelial cell dysfunction that proceeds atherosclerosis, depressed zinc status may be involved in the initiation of endothelial dysfunction. However, before recommending a zinc-enriched diet to reduce the risks for atherosclerosis, the effect of excess zinc on endothelial cell functions in normozincemic status should be known. Therefore, in this study, the effect of dietary zinc on normal endothelial cell functions in animals subjected to a diet containing 334 +/- 58 ppm zinc for 30 d was studied to see whether supplemented zinc has an effect on endothelial cells. Despite a slight increase in blood zinc, unaltered aortic and kidney zinc contents were associated with unchanged blood pressure in rats subjected to a zinc-enriched diet. Increased basal nitric oxide and prostacyclin were accompanied by a normal response to phenylephrine. Dietary zinc influenced neither endothelial-dependent nor endothelial-independent relaxations significantly. However, it elevated the share of M1-type cholinoceptor response as well as dilator prostaglandin release, which seems to be nitric oxide dependent. There was a strong correlation (r=0.826, p<0.05) between M1-type cholinoceptor response and prostacyclin release in zinc-treated rings. These results suggested that zinc ions increases M1 mediated prostacyclin release in normal endothelial cells without altering intracellular pathways. PMID- 12746574 TI - Age-dependent effects induced by ouabain in rat heart muscles and cellular Ca2+ concentration. AB - Responsiveness to ouabain of the inotropic and chronotropic effects in rat atrial muscles during development (3-18 wks old) was examined. In spontaneously beating rat right atrial muscles, ouabain (3-30 microM) caused a potent positive inotropic effect in a concentration-dependent manner, but failed to have a chronotropic effect; at 30 microM, 78.6 +/- 3.4% (n = 14, p<0.01) in the contractile force and -1.1 +/- 2.3% (n = 14, p>0.05) in the sinus rate in 10-wk old rats. The myocardium during development increased the responsiveness to ouabain (10 microM) by 27.6 +/- 2.1% (n = 14, p<0.01), 58.7 +/- 3.3% (n = 14, p<0.001), and 47.2 +/- 2.3% (n = 14, p<0.001) in 3-, 10-, and 18- wk-old rats, respectively. However, the response on the sinus rate was not modified in all of the developing stages. Higher frequencies of stimulation caused the more potent inotropic effect in left atrial muscles. In the experiments using a Ca2+ sensitive fluorescent dye (Fura-2), ouabain (10 and 30 microM) increased the cellular Ca2+ concentrations by 3.0 +/- 2.1% (n = 6, p>0.05) and 12.7 +/- 1.5% (n = 6, p<0.05) in 3-wk-old rats and by 13.0 +/- 2.7% (n = 6, p<0.05) and 42.9 +/- 3.1% (n = 6, p<0.01) in 18-wk-old rats, respectively. These results suggest that the ouabain-evoked response is enhanced during development (but tends to decrease from the maximum after maturing), presumably resulting from developmental degrees of cellular mechanisms such as Na+/K+ pump activity and Na+/Ca2+ exchange and is reflected by changes in the cellular Ca2+ concentration. PMID- 12746573 TI - Antioxidant properties of chromium and zinc: in vivo effects on digestibility, lipid peroxidation, antioxidant vitamins, and some minerals under a low ambient temperature. AB - The effects of chromium (chromium picolinate, CrPic) and zinc (ZnSO(4)H(2)O) supplementation on serum concentrations of malondialdehyde (MDA) (an indicator of lipid peroxidation) and serum status of some antioxidant vitamins and minerals of laying hens (Hy-Line) reared at a low ambient temperature (6.8 degrees C) were evaluated. One hundred twenty laying hens (Hy-Line; 32 wk old) were divided into 4 groups, 30 hens per group. The hens were fed either a basal diet or the basal diet supplemented with either 0.4 mg Cr/kg of diet, 30 mg Zn/kg of diet, or 0.4 mg Cr plus 30 mg Zn/kg of diet. Digestibility of nutrients (dry matter [DM], organic matter [OM], crude protein [CP], and ether extract [EE]) increased by supplementation of chromium and zinc (p < 0.05). Supplemental chromium and zinc increased serum vitamins C and E but decreased MDA concentrations (p < 0.05). Additionally, supplemental chromium and zinc caused an increase in the serum concentrations of Fe, Zn, Mn, and Cr (p < 0.05). The present study showed that low ambient temperature causes detrimental effects on the digestibility of nutrients and antioxidant status and that such detrimental effects caused by low ambient temperature can be alleviated by chromium and zinc supplementation, particularly when Cr and Zn were simultaneously included into the diet. Data obtained in the present study suggest that such supplementation can be considered as a protective management practice in a diet of laying hens for alleviating negative effects of cold stress. PMID- 12746575 TI - Dietary selenium levels determine epidermal langerhans cell numbers in mice. AB - Selenium (Se) is a dietary trace element that is essential for effective immunity and protection from oxidative damage induced by ultraviolet radiation (UVR). Langerhans cells (LC) represent the major antigen-presenting cells resident in the epidermis; a proportion migrate from the skin to the draining lymph nodes in response to UVR. Because it is known that Se deficiency impairs immune function, we determined what effect this has on LC numbers. CH3/HeN mice were weaned at 3 wk and placed on diets containing <0.005 ppm of Se (Se deficient) or 0.1 ppm of Se (Se adequate, control mice). After 5 wk on the diet, the epidermal LC numbers in the Se-adequate group were 966 +/- 51 cells/mm2 and LC counts in the epidermis of the Se-deficient mice were 49% lower (p<0.05). Glutathione peroxidase- I (GPx) activity was measured in the epidermis, lymph nodes, and liver. In the epidermis, the activity of GPx in the Se-deficient mice was only 39% (p<0.01) of that seen in epidermis from Se-adequate mice (1.732 U/mg protein). The mice were then irradiated with one dose of 1440 J/m2 of broadband UVB or mock irradiated. After 24 h, the decrease in LC number after UVB was greater in the Se-adequate mice, (40% decrease) compared to the Se-deficient group (10%). Thus, Se deficiency reduces epidermal LC numbers, an effect that might compromise cutaneous immunity. PMID- 12746576 TI - Zinc supplementation attenuates thioacetamide-induced liver injury and hyperglycemia in mice. AB - Zinc supplementation has been shown to improve not only liver dysfunction but also glucose intolerance in subjects with liver cirrhosis. In this study, we investigated the effects of zinc supplementation on the changes in circulating levels of tumor necrosis factor-alpha and total antioxidant capacity in mice with thioacetamide-induced liver injury. The protective effect of concurrent zinc administration for thioacetamide-induced hepatotoxicity was also examined. The results showed that zinc treatment significantly attenuated thioacetamide-induced liver injury and hyperglycemia. Furthermore, thioacetamide-induced hepatotoxicity was markedly weakened by the simultaneous zinc administration. These effects might be attributed to reduced tumor necrosis factor-alpha production and elevated total antioxidant capacity induced by the mineral. Our data suggest that zinc supplementation might be beneficial for the subjects with a high susceptibility to liver injury. PMID- 12746577 TI - Kinetics of the action of three selenides on Staphylococcus aureus growth as studied by microcalorimetry. AB - The effect of three kinds of selenide on Staphylococcus aureus growth was studied by means of microcalorimetry. Differences in their capacities to inhibit the metabolism of this bacterium were observed. The rate constant k (in the log phase) in the presence of the compounds decreased with increasing concentrations of the compounds. The relationship of k and c is nearly linear for the selenium compounds. Judged from the rate constant, k, and the half-inhibitory concentration IC50, the experimental results reveal that the sequence of antibiotic activity of the three tested selenides compounds is (2-hydroxy benzyl imino)ethyl n-hexyl selenide> n-butyl(2- hydroxy benzyl imino)ethyl selenide > bis[(2,4-dihydroxy benzyl imino)ethyl] selenide. PMID- 12746578 TI - Don't believe the hype: get SARS in proportion. PMID- 12746579 TI - The role of district nurses in public health: a discussion. AB - The government has challenged community nurses, through recent policy changes, to become more involved in improving the health of local populations. However, very little has been written about the role of district nurses within this agenda. This article explores these opportunities and discusses some of the challenges facing district nurses if they are to embrace this new agenda. A model is examined which details four approaches to public health in which district nurses could contribute: individual advice-giving, personal support, environmental control and community development. District nurses have many valuable skills and attributes that could be utilized within all four approaches; it is important that they use these skills in collaboration with others. There will be implications for the future education and leadership of district nurses. PMID- 12746580 TI - Suprapubic catheter removal: the cuffing effect of deflated catheter balloons. AB - Supra-pubic catheterization plays an important role in patient care and management when this method of indwelling catheterization is required. However, one area of concern often experienced by nurses is the problem removing supra pubic catheters or not being able to remove it. Catheter balloons, when deflated, incur crease or ridge formation. Removing supra-pubic catheters, a 'cuffing' effect occurs as the catheter is being removed. This seems to affect 100% silicone catheters more than non-silicone catheters. This article looks at the changes 100% silicone catheter balloons undergo following deflation and removal. PMID- 12746581 TI - Patient group directions for antibiotic supply in Walk-in Centres: how legal are they? AB - Until recently antibiotics were only available on prescription. Nurses in NHS Walk-in Centres (WiCs) now supply and administer antibiotics in accordance with patient group directions (PGDs). The aim of this study was to investigate the process of antibiotic supply from WiCs. Antibiotic PGDs from ten WiCs were selected and medical notes of 50 patients who received an antibiotic at these WiCs were examined to determine compliance with PGD specifications. No consistency of approach in format and content of PGDs was found. PGDs from six WiCs complied with 15/20 or fewer PGD requirements. In the examination of patients' records, 65% of all relevant PGD requirements were explicitly recorded. The breadth and variability of PGDs demonstrates local needs and priorities; however, low compliance levels with PGD requirements provide cause for concern. This article discusses the legality of PGDs, equity in both service provision and advice offered, compliance of medical notes with PGD requirements, implications for patients and WiC nurses, and training implications. Incomplete documentation makes it impossible to verify the quality of care provided across and within WiCs. It also suggests that uncertainty remains around the supply of medicines under PGDs. PMID- 12746582 TI - Preventing falls in older people: risk factors and primary prevention through physical activity. AB - Falls among older people represent a major public health issue, which can in part be tackled through an integrated falls service combining both primary and secondary prevention. Many falls can be prevented following comprehensive assessment to identify risk factors and to plan interventions to eliminate them or ameliorate their effect. Community nursing staff are well placed to undertake such risk assessments and can instigate programmes of primary prevention designed to reduce the likelihood of a person falling. Increased physical activity among older people represents one element of a prevention programme. While this is beneficial for the older person's general health and well-being, certain types of exercise can also be used to reduce falls in individuals with muscle weakness, reduced mobility and balance problems. With the exception of balance training the evidence base related to exercise and falls prevention is patchy; Carter et al (2001) suggest that as yet there is insufficient evidence to suggest an optimum exercise programme for falls prevention. Each person should therefore be individually assessed and the results used to identify what type of exercise they might benefit from in order to address a specific risk factor. Once an appropriate form of exercise has been identified, practitioners should put the older person in contact with a physical activity coordinator to assist them in accessing an exercise programme. PMID- 12746583 TI - Fluticasone and beclometasone: what are their effects on children's growth? AB - Fear of growth retardation may account for the underuse of inhaled corticosteroids in children with asthma, despite compelling evidence of their effectiveness. This fear may be reduced with newer agents with lower oral bioavailability if their theoretical advantage of fewer systemic adverse effects than the standard treatment of inhaled beclometasone is realized in practice. This review aims to determine if one of the newer agents, inhaled fluticasone, has less effect on the growth of pre-pubertal asthmatic children than inhaled beclometasone. The outcome measure was growth velocity. Two double blind, randomized controlled trials were identified. In one of the studies the mean growth velocity in the fluticasone group was 0.7 cm/year greater than in the beclometasone group. In the second, smaller study the mean growth velocity in the fluticasone group was 0.8 cm/year greater. There is therefore some evidence that fluticasone has less (if any) adverse effect on growth. PMID- 12746584 TI - An evaluation of clinical tools to measure pain in older people with cognitive impairment. AB - Pain in older people with cognitive impairment is often untreated or under treated despite the fact that the likelihood of having pain increases with advanced age. One reason that pain is poorly managed in older people is that it is not detected. Assessing pain is an integral part of clinical practice and is required for effective pain management. However, systematically measuring pain intensity and location with standardized tools in older people has not been well studied. This article will summarize tools used to assess pain in older people, inducing self-report scales measuring pain intensity, the presence or absence of pain, pain location and pain behaviour scales. Identifying the advantages and disadvantages of these clinical assessment tools has implications for nursing practice, research and education. PMID- 12746585 TI - All epidemics deserve health care attention. PMID- 12746587 TI - It is time to really value care workers for the elderly. PMID- 12746586 TI - RCN votes that nursing should not become all-graduate. PMID- 12746588 TI - Nurse in bad health who did not know who to turn to for help. PMID- 12746589 TI - Medicines and the older person: principles of good practice. AB - The UK has an ageing population--there are currently more people aged over 60 years than aged 16 years and under. The National Service Framework (NSF) for Older People provides standards for health and social services to ensure that older people receive high quality care (Department of Health (DoH), 2001a). An addendum to the NSF describes how the use of medicines for and by older people can be improved (DoH, 2001b). The aim of the document is to ensure that older people achieve the greatest benefit from their medications in order to sustain and increase their quality of life. The document also attempts to ensure that the older person does not suffer needlessly from illness that may be brought about by excessive, inappropriate or inadequate consumption of medicines. This article discusses how medications for older people can be taken in a safe and effective manner in relation to the addendum to the NSF (DoH, 2001b). Nurses must ensure that a multidisciplinary approach is adopted when setting standards and developing protocols for the safe and effective administration of medicines. PMID- 12746590 TI - Interpretation of terms used to describe handwashing activities. AB - A literature review on the use of alcohol-based handrubs highlighted many variations in the terms used to describe the activities related to effective hand hygiene. This suggests that an individual's interpretation of the terms used to describe hand-hygiene activities may lead to a variation in compliance with any protocol or guideline employing them. This article describes an audit that was undertaken to establish whether the definition of the terms handwashing, hygienic handwashing and hygienic handrub varied between grades of staff and disciplines who have direct contact with patients. The aim was to identify different healthcare professionals' understanding of the terms used to describe hand cleansing activities. The results indicate that a variation in understanding of these terms did exist between these grades and disciplines. This may indicate that there is a need for the definition of the terms used to be clearly stated, consistent in their use and universally adopted. PMID- 12746591 TI - Living theory: enhancing the psychological support of patients. AB - This article explores how nurses, involved in a research project incorporating teaching psychological theories and counselling skills to enhance the psychological support of patients with wounds, had, one year on, changed their professional practice. This inquiry was framed by living theory, a concept previously only used in education, which is based on the integration of known knowledge, newly taught knowledge and increased self-awareness. The major principle of living theory is that one's values are questioned, modified, clarified and sometimes changed completely in striving to improve one's professional practice. This research showed nurses creating their own living theories, aspiring to really care for the whole person by developing strong, meaningful relationships with patients. The steps that participants took from first using the enhanced way of working with patients with wounds, to using it to support all patients psychologically, are demonstrated. PMID- 12746592 TI - Case study of alopecia universalis and web-based news groups. AB - This article presents findings from an 18-month case study of a web-based news group used by individuals with alopecia universalis. Content analysis of 228 episodes of web-based communication that occurred in relation to themes of discussion was undertaken, supported by the use of concept mapping (Northcott, 1996). Analysis identified a core concept relating to that of a community of shared experience together with four supportive themes. The themes were the search for understanding and meaning, carrying on, seeking balance between past, present and future, and relating to new self, others, and the world. The article discusses the increased growth in the use of the web as a vehicle for exploring health concerns and the specific ethical and methodological issues raised by research in this area. PMID- 12746593 TI - Legal regulation mechanisms in the control of medicines. AB - In 2002, a case was brought against the Secretary of State--sued on behalf of the Committee for the Safety of Medicine (CSM)--by a mother who in May 1986 gave aspirin to her 6-year-old daughter who was suffering from chickenpox (Smith vs. Secretary of State for Health, [2002]). The child deteriorated and following admission to hospital she was diagnosed as suffering from Reye's syndrome. As a consequence, the girl was left with a serious neurological handicap and was totally dependent on others for her care. In June 1986, the government issued a general public warning advising parents not to give aspirin in any form to children under 12 years old and told chemists to take all junior aspirin preparations off their shelves. This followed advice from the CSM that in this age group aspirin could trigger the fatal disease Reye's syndrome. The CSM had been aware of this danger in September 1982 following evidence in the USA. Further evidence was produced in 1985 and the CSM continued to monitor the situation. In April 1986, a meeting was held between the Department of Health (DoH) and members of the aspirin industry. It was subsequently agreed that the producers would withdraw stocks and there would be cooperation between the industry and the DoH rather than a public warning. In May 1986, the CSM recommended that the DoH should give a general public warning, which was made on 10 June 1986. The mother's case was that there was an unreasonable delay in publishing a warning following the CSM meeting on 26 March 1986. It was accepted that the administration of aspirin was a contributory factor in the development of the girl's neurological condition and that had her mother been aware of the warnings before May 1986 she would not have given her aspirin. Were the Secretary of State and CSM liable? PMID- 12746594 TI - Pressure ulcer prevention: education for nursing home staff. AB - This article describes an education programme for a group of nurses working in several nursing homes located in different areas of the Midlands but each belonging to the same care group. The group's management team had identified that there were patients in the nursing homes who had severe pressure ulcers and that staff were not managing their care adequately in order for healing to occur. It has been identified that 'education is probably the single most effective way of reducing the incidence of pressure ulcers' (Department of Health (DoH), 1993). Although the various nursing homes were able to access the skills of clinical nurse specialists in tissue viability, severe pressure ulcers were failing to heal and nursing home staff requested additional education to help them address this problem. Nurses in the homes expressed a desire to gain a deeper knowledge of the problem, so they would be able to plan and implement appropriate care autonomously and thus raise the standard of pressure ulcer care provided in each home. This article discusses the implementation of a comprehensive education programme that contributed to raising the standards of patient care and to the professional self-worth of the nurses involved. PMID- 12746596 TI - It is important to support nurses in trouble. PMID- 12746595 TI - A new modern drainable appliance for people with ileostomies. AB - Finding the right drainable pouch can be difficult for the person with an ileostomy. There is an extensive range of appliances available on prescription that offer a variety of features and benefits. Welland Medical has introduced a new one-piece drainable pouch that has a secure integral locking device on the outlet. This new pouch also incorporates a unique Dual-Carb filter which combines modified and unmodified carbon to help fight the various odour types that an ileostomy can produce. PMID- 12746597 TI - [Non-biodegradable drug-sustained capsular ring for prevention of secondary cataract. Part I: In vitro evaluation]. AB - PURPOSE: The appearance of secondary cataract constitutes the most common complication of cataract surgery since the advent of phacoemulsification. Prevention is today one of the most crucial challenges of research in ophthalmology. We have developed a drug-sustained capsular ring releasing 5 fluorouracil (5-FU). This system aims to combine mechanical and pharmacological properties with the ability to interfere on the lens epithelial cellular processes of secondary cataract. This first article will report on the in vitro evaluation of this system. MATERIALS AND METHOD: Development of a hydrogel that can serve as a mechanical support and as a release of an active agent requires a comparative trial of multiple hydrophobic products providing rigidity to the ring and of hydrophilic products providing the ability to release a drug. The choice of 5-FU as a non-proliferate agent was based on its well-known pharmacological properties and its wide use in ophthalmology. Evaluation comprised adjustment of the adequate copolymer, then of the loading phase, and finally assessment of 5-FU release. The mechanical characteristics and cytocompatibility of the ring were also assessed. RESULTS: The characteristics of drug release differed depending on the water content and resulting volume of the copolymer. The pharmaceutical and mechanical testing led to selecting the HPMA-MMA combination (75:25). However, a burst effect within the first 2 hours was observed in all cases. That limit was eliminated by creating a biodegradable surface coating of Poly Lactid-co-Glycolid (PLG 50:50) that could provide a positive barrier effect against migration and a controlled continual release of 0.25 microg/h extended to 9 days. Mechanical tests of tension, compression, and traction showed the crucial influence of the ring's hydration state. The cytocompatibility study showed no evidence of cellular toxicity. DISCUSSION/CONCLUSIONS: The synthesized copolymer is able to release a constant, regular and safe rate of 5-FU, higher than CI50 over a period of 9 days for a cell proliferation phase of 3-5 days. We adjusted the ring associating the mechanical properties and the ability to release 5-FU, allowing further evaluation on animal models, the subject of the second part of this report. PMID- 12746598 TI - [Disinfection of gas-permeable contact lenses against prions]. AB - Potential iatrogenic transmission from patients incubating Creutzfeldt-Jakob disease, especially variant CJD, is a major public health issue. Because the ocular route is very efficient for contamination with prions, re-use of rigid contact lenses in ophthalmology constitutes a potential problem. We therefore evaluated the anti-prion activity of different protocols available for disinfection of lenses. These treatments decreased the infectivity retained on the surface of experimentally contaminated lenses by a factor of at least 10 million. They thus represent an important factor in protecting against possible prion infection via the ocular route. PMID- 12746599 TI - [Status of low-vision rehabilitation for age-related macular degeneration by orthoptists in the North of France]. AB - AIM OF THE STUDY: To evaluate the status of low-vision rehabilitation carried out by the orthoptists in the North of France. This study was a preliminary step in setting up a network between different vision professionals for the management of age-related macular degeneration (AMD) the North of France. METHODS: The ASO (Association Septentrionale d'Ophtalmologie) conducted a survey funded by the URCAM (Union Regionale des Caisses d'Assurance Maladie) with the FAQSV (fond d'aide a la qualite des soins de ville). The survey was based on the analysis of two forms sent to orthoptists of the North region of France. RESULTS: The survey analyzed 46 responses (a representative sample with 69% responses) providing a description of the orthoptists of the North of France: 19% males, 81% females whose mean professional experience was 13.3 years. Thirty-four percent of the orthoptists have had training in low-vision rehabilitation. This training was given during the university courses for 21% of responders and during a postgraduate course for 79%. Of the orthoptists surveyed, 64% worked in a private context, 9% in a public context, and 27% in both public and private contexts. Their main activity was in their own private practice for 60%, in an ophthalmologist's office for 20%, in a public institution for 16%, and a private institution for 4%. The mean number of patients treated was 70 per week per orthoptist, with 21% working mostly with children, 36% working mainly with adults between 16 and 60 years of age, and 7% with the elderly, whereas 36% reported no specificity related to patient age. The mean number of patients dealt with for low vision related to AMD was 4.1 per month per orthoptist. The average number of patients dealt with for low vision with no relation to AMD was 1.5 per month. The prescriber of low-vision rehabilitation was an ophthalmologist for 88.9% of the orthoptists and a general practitioner for 11.1%. Questions addressed to AMD patients: at the beginning of the survey, 83.8% of the patients did not have sufficient visual acuity to be able to read a text of current size (Parinaud 4); 40.4% of the patients required help for everyday life, and 59.6% were autonomous. For 7.1% of the patients, low-vision rehabilitation was carried out less than 1 month after the stabilization of retinal lesions, but in 35.3%, rehabilitation was carried out more than 2 years after lesions were stabilized. The main request of the patients involved improvement of near vision (89.9%). CONCLUSION: This survey will be a preliminary step in setting up a regional health network coordinating the ophthalmological and orthoptic management of AMD. PMID- 12746601 TI - [Bacterial contamination: epidemiology in cataract surgery]. AB - PURPOSE: To evaluate anterior chamber (AC) bacterial contamination at the end of cataract surgery in a large series of patients, to determine the influence of operative technique on ocular contamination. METHODS: Retrospective study of 2,624 patients undergoing cataract extraction, 354 extracapsular cataract extraction (ECCE) and 2,270 phacoemulsification. Anterior chamber aspirates were performed on completion of surgery for microbiological studies. RESULTS: One hundred and thirty two patients (5%) had culture-positive anterior chamber aspirates. Coagulase-negative Staphylococcus, Propionibacterium sp. and Corynebacterium sp. were the most commonly isolated organisms. The AC contamination rates during ECCE (5.6%) and phacoemulsification (4.7%) were not statistically different. There was a statistically significantly higher risk of AC contamination in eyes receiving an intraocular lens (IOL) with polypropylene haptics (9.9%) than in eyes receiving the same IOL with polymethylmethacrylate haptics (4.4%). CONCLUSION: Surgical technique had no statistically significant effect on ocular contamination. Polypropylene haptics IOLs were associated with a higher risk of bacterial contamination. PMID- 12746600 TI - [Retinal vein occlusion and hyperhomocysteinemia]. AB - INTRODUCTION: Previous studies have reported that an elevated plasma homocysteine level is a risk factor for vascular disease. The aim of this study is to determine whether hyperhomocysteinemia is a risk factor for retinal vein occlusion (RVO) and whether it is a prognostic factor. PATIENTS AND METHODS: The plasma homocysteine level was measured in 101 patients and compared to the plasma homocysteine level of controls. The relation between plasma homocysteine level and the other known risk factors of retinal vein occlusion was studied, as well as the correlation between the clinical outcome of the RVO and the plasma homocysteine level. RESULTS: The mean plasma homocysteine level was significantly higher in the 101 RVO patients than in the 29 controls (11.9 mmol/l vs 8.6, p<0.001). We found no relation between plasma homocysteine and other risk factors of vascular disease except for the hematocrit level. Hyperhomocysteinemia was more frequent in the ischemic forms and in bilateral RVO, but the difference was not statistically significant. CONCLUSIONS: Hyperhomocysteinemia seems to be an independent risk factor for RVO and was more frequent in severe RVO, but our study did not evidence an association with a severe prognosis. Vitamin therapy can decrease homocysteinemia but its efficacy in the prevention and in the treatment of RVO remains to be demonstrated. PMID- 12746602 TI - [A new model of induced ocular hyperpressure using the minipig]. AB - PURPOSE: To characterize a new animal model of moderate chronic hyperpressure obtained by obstruction of the iridocorneal angle (ICA) in the minipig. METHODS: Intraocular hyperpressure was induced in one eye (left) using an injection of methylcellulose (4%) in the anterior chamber of six healthy adult minipigs. Intraocular pressure (IOP) was measured before injection and at D+60 and D+180. The clinical condition thus created was regularly assessed with the following procedures: fundus photography, electroretinography (ERG) to evaluate retinal function, scanning laser ophthalmoscopy (SLO) angiography to measure the arteriovenous filling times (AVFT). Optical microscopy was also performed to evaluate iridocorneal angle and inner retinal layers. RESULTS: In all instances the injection produced a significant increase in the IOP accompanied by a mydriasis, as well as a significant increase in the AVFT and reduction (abolition in some cases) in the i-wave of the ERG. Fundus examination also revealed a blurred aspect and reduction in the calibre of the retinal blood vessels. Similarly, all experimental eyes showed, at optical microscopy, obstruction of the ICA as well a significant loss of of retinal ganglion cells. CONCLUSION: Our results suggest that the above pathophysiological processes, triggered by the induced hyperpressure, share many similarities with human chronic open-angle glaucoma. Consequently, our model, which is very easy to create, could be used to test new therapeutic agents such as neuroprotective drugs. PMID- 12746604 TI - [Melanoma of the iris and pregnancy]. AB - Iris melanoma is a malignant melanocytic tumor, making up 1.2%-6.6% of uveal malignant melanomas. The growth of choroidal melanoma during pregnancy is described and a hormonal influence is suggested. We present a case of iris melanoma occurring during pregnancy. A 32-year-old woman, in the 28(th) week of her fifth pregnancy, was referred to the ophthalmologist with a 2-month history of blurred vision and pain in her left eye. The visual symptoms became progressively worse with decreasing vision. Her visual acuity was 20/70 in her left eye, with a temporal, pigmented, prominent iris mass extending into the trabecular meshwork and the corneal endothelium. Elsewhere other pigmented localizations were found in the iris and in the trabecular meshwork. There was secondary glaucoma with intraocular pressure of 36mmHg and C/D=0.9. The right eye was normal. A general physical examination found no pigmented lesions. Chest radiography and hepatic ultrasonography revealed no metastasis. The diagnosis of iris malignant melanoma was made from the diffuse involvement extending into the trabecular meshwork and the advanced secondary glaucoma; an enucleation was performed and histopathological examination confirmed the diagnosis of iris melanoma with involvement of the trabecular meshwork. The patient had no further problems (follow-up of 11 months). From this case report, we discuss the differential diagnosis of iris melanoma and the hormonal influence on its growth. PMID- 12746603 TI - [Treatment of uveal melanoma with iodine 125 plaques or proton beam therapy: indications and comparison of local recurrence rates]. AB - INTRODUCTION: This retrospective study compared the rate of local recurrence after irradiation of uveal melanoma treated with iodine 125 plaques or proton beam therapy. PATIENTS AND METHODS: Iodine 125 plaques were used to treat all uveal melanomas between the end of 1989 and 1991. Since 1991, we have used iodine plaques for small anterior tumors and proton beam for other tumors. We use a plaque with a larger diameter than the tumor diameter (2-4mm) with a dose of 90Gy at the apex. Proton beam therapy is used for all tumors at the equator or posterior to the equator not thicker than 12mm. The dose given is 60Gy cobalt equivalent in four fractions. For each patient, the initial size and location of the tumor were noted as well as the follow-up each year: the outcome for the eye (local recurrence, ocular conservation, and functional results), the occurrence of metastasis, and survival. A statistical analysis was performed. RESULTS: Between December 1989 and September 1998, 1272 patients were treated: 926 (72.8%) were treated with proton beam irradiation and 346 (27.8%) with iodine 125 plaques. The median follow-up was 5 years (60 months). For the patients treated with proton beam therapy, the mean age was 58 years, the tumor location was anterior to the equator for 3.8%, at the equator for 43.6%, and posterior to the equator for 52.6%. The mean tumor diameter was 13.4mm and the mean tumor thickness was 5.69mm. For the patients treated with iodine 125 plaques, the mean age was 61.5 years. The location of the tumor was anterior to the equator for 34.4%, at the equator for 46.5%, and posterior to the equator for 19.1%. The mean tumor diameter was 11.5mm and the mean tumor thickness was 5.12mm. The recurrence rate was 4% for the proton beam treatment and 3.75% for iodine plaques. There was no statistical difference. DISCUSSION: In the literature, the rate of local recurrence is usually higher with iodine 125 plaques than proton beam therapy. We discuss the risk factors for local recurrence after iodine 125 plaques: tumor diameter, lower dose to the tumor apex and lower dose rate, and posterior location of the tumor. We found a higher mortality rate in patients who presented local recurrence. CONCLUSION: When we use iodine 125 plaques for anterior tumors with the proper dose and dose rate to the apex of the tumor, we do not find more recurrence than with proton beam therapy. PMID- 12746605 TI - [Diagnosis of paroxysmal episodes of eye rising in a child]. AB - This report presents a case of a 7.5-year-old boy who was given a complete check up with normal results for paroxysmal elevation of the eye with onset during seizures. The clinical aspect showed a tonic upgaze that has never been described in such an old patient. We describe this rare anomaly and review the literature. The main differential diagnosis was the presence of a tic. PMID- 12746607 TI - [Ciliary body melanoma in a black patient: a case report]. AB - Uveal melanomas are rare in the black population. We report the case of an 81 year-old black man who presented a ciliary body melanoma of the left eye. Ophthalmological examination revealed a mass arising from the ciliary body and choroid. Diagnosis of ciliary body melanoma, made from clinical examination and ocular ultrasound, may be difficult in some cases due to its similarity to other eye tumors. Uveal melanomas mainly affect Caucasians but they are not unknown in blacks. PMID- 12746606 TI - [A new case of Susac syndrome and a review of the literature]. AB - We report a single case of Susac syndrome (microangiopathy of the brain, retina, and cochlea). A 26-year-old woman developed branch retinal artery occlusion in the right eye, associated with bilateral hearing loss that mostly involved low frequencies. MRI of the brain revealed small multifocal hyperintensities in the white matter of the cerebrum on T2-weighted images with gadolium enhancement. The treatment consisted of anticoagulation and antiplatelet drugs. Seventy-one cases of Susac syndrome have been reported in the literature. The Susac syndrome is more frequent in females and its etiology remains unknown. However, immune inflammatory disorders, vasospastic phenomena, and coagulopathy could be involved in its pathophysiology. Treatment options are not standardized, ranging from antithrombotic drugs to immunomodulatory therapy. The course of the disease is self-limited after an initial fluctuating active phase. The prognosis of Susac syndrome is good in most cases. PMID- 12746608 TI - [Molecular control of apoptosis]. AB - Apoptotic cell death is a natural event necessary to shaping the developing nervous system and is a feature of neurodegenerative disease pathology. Subtle interactions between pro- and anti-apoptotic molecules are controlled by environmental factors such as trophic factors. The mitochondrion is a major component regulating these interactions. At the time of apoptosis, proteases, called caspases, are activated to ensure cell breakdown. In living cells, intracellular components ensure the inhibition of caspases. As such, caspases are therapeutic targets to induce or to prevent apoptosis. PMID- 12746609 TI - [Apoptosis and the ocular surface]. AB - Apoptosis, or programmed cell death, is an active phenomenon that plays a major role in most mechanisms of regulation, differentiation and wound healing. Mostly studied in the retina, apoptosis is also extensively involved in the anterior segment, especially the ocular surface. Apoptosis of keratocytes is a rapid phenomenon following excimer refractive surgery. Any epithelial aggression stimulates a series of mechanisms leading to death of deep keratocytes. The role of epithelial cell mediators may explain the superiority of LASIK compared to PRK in terms of functional rehabilitation. Conjunctiva is also a major site in which inflammation and apoptosis are combined. Proinflammatory cytokines may both amplify immune reactions and stimulate epithelial apoptosis, which is most likely to result in elimination of injured tissues. Toxic drugs also play a major role and iatrogenic apoptosis should be avoided as much as possible, especially by eliminating preservatives from eyedrops, most of which use both proinflammatory and proapoptotic agents. PMID- 12746610 TI - [Age-related macular degeneration and apoptosis]. AB - Several studies suggest that apoptosis might play a major role in age-related macular degeneration. Apoptosis in retinal pigment epithelium cells undergoing severe oxidative stress has been reported and could therefore be involved in the pathogenesis of AMD. The processes of drusen formation seem to be similar to the processes described in apoptosis. Moreover, apoptosis appears to be involved in the early outgrowth of choroidal neovascular membranes as well as in the development of fibrotic scars at a later stage. Targeting apoptotic pathways should therefore be considered as a possible treatment approach for AMD. PMID- 12746612 TI - [Iris atrophy]. PMID- 12746613 TI - [Hypertrophic eyelid conjunctival scar. A tarsal keloid]. AB - The clinicopathologic case of a 53-year-old female patient with an abnormal tumor growing on the mucous part of the superior right eyelid is reported. The patient was operated on for ten years ago and a whitish mass slowly developed on the conjunctival face of the eyelid disturbing the use of corneal lenses. It was hard, painless and had the shape of a flat mushroom. The removal was performed under local anesthesia and allowed us to resect a hard and fibrous lesion. Histopathology showed that the lesion was made of a fibrous tissue organized like a hypertrophic scar. Surgical treatment of chalazion is frequent and rarely gives rise to abnormal scarring. PMID- 12746615 TI - [Early stages of beta-cell deficiency: the anticipated chronicle to type 2 diabetes]. AB - Type 2 diabetes combines two mechanisms, peripheral insulin resistance and beta cell defect. In this disease, insulin secretion is rather difficult to evaluate because: 1) the multiplicity of tests exploring different aspects of insulin secretion, 2) the complex relationships between beta-cell function and either insulin resistance or plasma glucose level. In glucose tolerant populations at risk of diabetes, insulin secretion has been found heterogeneous with large inter individual variations. However, insulin secretion is decreased in almost 50% of the studies. In glucose intolerant subjects, insulin secretion is more consistently altered, specially acute insulin response to glucose. More subtle abnormalities, as decrease of proinsulin cleavage or pulsatility, are rare at this stage. When diabetes is diagnosed, beta-cell defect is obvious whatever the test used, and progress with time. The markers of the transition to glucose intolerance are both insulin resistance and acute insulin response. The transition to diabetes is associated with the same factors plus the dysregulation of hepatic glucose production. Hyperinsulinism and high proinsulin/insulin ratio are good predictors of diabetes in at risk populations. Loss of beta-cell function is a major factor in the progressive worsening of the disease, specially in glucose intolerant subjects. PMID- 12746614 TI - [Metabolic syndrome: to observe or to act?]. AB - Type 2 diabetes is characterized by a very high cardiovascular risk. This risk has usually been present for a very long time when diabetes is diagnosed. The combination of several metabolic abnormalities known as metabolic syndrome seems to be responsible for this increased risk. Insulin resistance is a major component of metabolic syndrome, which also includes clinical abnormalities such as increased waist circumference, hypertriglyceridemia, high blood pressure. It is very important to screen subjects with metabolic syndrome because of their high risk. Furthermore, some pharmaceutical agents could be effective in reducing the risk related to insulin resistance. Metformin effectiveness has been shown for the reduction of diabetes related complications and mortality, mainly in the cardiovascular field. Its place among the other potentially active agents remains to be determined as well as how it should be used in combination to lifestyle changes and how effective it is on a long-term basis. PMID- 12746616 TI - [Implementation of the "Diabetes Prevention Program", in Europe]. AB - Prevention of type 2 diabetes has been a major issue for healthcare providers because of the high incidence and the high cost of diabetes. High cost is related to diabetes treatments, diabetes complications and the reduction of lifetime expectancy. The Diabetes Prevention Program study has recently shown that the prevention of diabetes is possible in patients with impaired glucose tolerance. In this study, diabetes was prevented either by metformin or by intensive lifestyle changes. Intensive lifestyle changes were even more effective than metformin in preventing diabetes. This article discusses how difficult it would be to implement this program in Europe. The difficulties would be both practical (identification of the high risk population, patients' motivation for intensive lifestyle changes) and economic. The implementation of the program in Europe would be theoretically cost-effective but would be technically very hard to do. PMID- 12746617 TI - [Understanding patients to promote self-regulation in Type 2 diabetes: how to live with an illness beginning before its onset?]. AB - The characteristics of Type 2 diabetes raise major questions on patients' behaviours and their determinants. The goal of this paper is to integrate recent empirical results of behavioural sciences and modern medical therapeutics in diabetes. We should consider all factors which may influence behavioural self care while refer less to the normative logic of compliance. Behavioural sciences have demonstrated that various central factors should be considered to promote health behaviours in Type 2 diabetes: namely negative emotions, coping, personal models of illness and risk perception. All these concepts may constitute targets of practical interventions. These can either improve the quality of life, modify or individualise diabetes constraints, improve the focus of medical information given to the patient, favour a better acceptation of illness or a more active role towards diabetes. Self-monitoring of blood glucose and introduction of insulin may influence psychological determinants of self-care behaviours. These arguments are followed by a set of recommendations for the clinician and the researcher. PMID- 12746620 TI - [History, accuracy and precision of SMBG devices]. AB - Self-monitoring of blood glucose started only fifty years ago. Until then metabolic control was evaluated by means of qualitative urinary blood measure often of poor reliability. Reagent strips were the first semi quantitative tests to monitor blood glucose, and in the late seventies meters were launched on the market. Initially the use of such devices was intended for medical staff, but thanks to handiness improvement they became more and more adequate to patients and are now a necessary tool for self-blood glucose monitoring. The advanced technologies allow to develop photometric measurements but also more recently electrochemical one. In the nineties, improvements were made mainly in meters' miniaturisation, reduction of reaction time and reading, simplification of blood sampling and capillary blood laying. Although accuracy and precision concern was in the heart of considerations at the beginning of self-blood glucose monitoring, the recommendations of societies of diabetology came up in the late eighties. Now, the French drug agency: AFSSAPS asks for a control of meter before any launching on the market. According to recent publications very few meters meet reliability criteria set up by societies of diabetology in the late nineties. Finally because devices may be handled by numerous persons in hospitals, meters use as possible source of nosocomial infections have been recently questioned and is subject to very strict guidelines published by AFSSAPS. PMID- 12746618 TI - [Drug compliance in type 2 diabetes: role of drug treatment regimens and consequences on their benefits]. AB - Compliance is an old issue but crucial in the management of chronic diseases. This is the case in type 2 diabetes mellitus which requires several drugs, either to treat diabetes or to prevent cardiovascular complications. In this review, we discuss the relationships between drug treatment regimens and treatment compliance in type 2 diabetes and other chronic diseases. The greater the number of daily drug intakes, the worst the compliance, even if a single daily intake may cause an increased risk of overdosing. Although the number of tablets or treatments is less frequently linked to compliance level than the number of daily intake, polytherapy is generally associated with a poor compliance. The consequences of a poor compliance on the prognosis or the management of these diseases are analysed based on cardiovascular studies. Even if nearly no studies exist in type 2 diabetes, to improve treatment compliance represents a major challenge in these patients. Such improvement requires to preferentially use once a-day intake, but this is still difficult with several oral anti-diabetics. Fixed combinations such as the glibenclamide plus metformin combination, cause a decrease in the number of daily tablets and this permit a better compliance. Such approaches, to be fully beneficial, should be part of a global management of these type 2 diabetic patients, taking into account all their difficulties to follow their treatments, and based on a strong physician and patient relationship. PMID- 12746621 TI - [Continuous glucose monitoring: different systems, different ambitions]. AB - This paper describes the current approaches towards the development of continuous glucose monitoring system, in which glucose is measured either in blood or in the subcutaneous tissue. These systems may be used as a glycemic holter, or to display continuously the changes in glucose level, to trigger an alarm in case of hypoglycemia, or of risk of hypoglycemia, or finally to pave the way to the development of an artificial beta cell. PMID- 12746622 TI - [Insulin therapy and self-monitoring of blood glucose: therapeutic management and recommendations]. AB - Self-monitoring of blood glucose (SMBG) has been widely spread during the last twenty years, and is at present considered as mandatory for the management of insulin therapy. The ALFEDIAM and the ADA recommend type 1 diabetic patients to perform more than 4 capillary glucose determinations per day, ideally 4 to 6. To be efficient in using these results, several parameters are required: glycemic targets have to be clearly defined, an intensified insulin regimen (3 or 4 injections per day or insulin pump therapy) should be proposed, and the patient should be teached how to modify his insulin doses. Short acting insulin analogs are available, and long acting insulin analog will be soon. This is a real progress in the choice of flexible insulin regimen. The practice of SMBG and its use for insulin adjustment requires continuing education, in order to translate SMBG in real diabetes self-management. This prerequisite is mandatory to obtain a sustained improvement in metabolic control. PMID- 12746623 TI - [Self-monitoring of blood glucose in type 2 diabetic patients. What could we propose according to their treatment?]. AB - Self-monitoring of blood glucose (SMBG) is well acknowledged in type 1 diabetes mellitus (DM). However, although its use is widely recommended in some developed countries, with a significant economical cost, SMBG remains questionable in type 2 DM because its efficacy is not demonstrated by high-quality randomized controlled trials. Type 2 diabetes is an heterogenous disease and its natural history covers several decades. Although type 2 DM represents an important risk of microvascular and cardiovascular complications, it remains asymptomatic for a long time with often a very poor metabolic control and low drug compliance. Treatment of type 2 diabetes varies during these decades, changes in lifestyle at the very beginning, then using oral drugs without risk to develop hypoglycemia (metformin, glitazones, alpha-glucosidases inhibitors) and/or insulin secreting drugs (sulfonylureas or glinides) with potential hypoglycemic risks. At every stage, SMBG could sensitize the patient and possibly could improve glycemias and compliance. SMBG must be adapted to provide informations representing a very useful, motivating and pedagogic tool. Obviously SMBG, in non-insulin-treated type 2 DM, targets other objectives when compared to its use in type 1 DM requiring adapted educational programs to avoid its uselessness and depression of the patient. In insulin treated type 2 DM patients, with a bed time regimen, SMBG represents both a major tool for adjusting the insulin and oral hypoglycemic drugs doses and a comprehensive instrument for controlling and understanding the disease. Thus SMBG is a logical tool for the management of a large proportion of type 2 DM patients but it requires to be proposed in structured counseling educational programs adapted to the psychological and social profile of the patients. These programs must be evaluated by randomized controlled trials. PMID- 12746624 TI - [Today data management in self-monitoring of blood glucose for diabetic patients]. AB - Improving diabetes treatment needs intensive glucose monitoring which is restricting for patients and time-consuming for physicians. Up-to-date tools of data management were developed, following progress in computing technology and home computing. Glucometers with memory and softwares are able to improve data management of self blood glucose monitoring, personalized interactivity with physician. They are very important to develop telemedecine systems in diabetes care. These systems are designed to complement the daily care and intensive management of diabetics through telemonitoring and telecare services. PMID- 12746625 TI - [Self-monitoring of blood glucose in gestational diabetes mellitus]. AB - About 6% of all pregnancies are complicated by gestational diabetes mellitus in France. Consensus is obtained for defining but not for the screening and diagnosis criteria. Management of gestational diabetes mellitus reduce materno foetal morbidity. Postprandial blood glucose peaks are highly predictive of macrosomia and have to be controlled in strict rates: less than 1.40 g/liter one hour after meal and less than 1.20 g/liter two hours after meal. Self-monitoring of blood glucose allows to start more rapidly insulintherapy, to obtain a better glycemic control and to reduce macrosomia. The impact of self-monitoring appears to be lower in the management of diet-controlled gestational diabetes. After delivery, self-monitoring of blood glucose can be used to control normalisation of blood glucose of women with high risk of type 2 diabetes mellitus. PMID- 12746626 TI - [Self-monitoring blood glucose in type 2 diabetes: for whom? And why? Which proofs?]. AB - Though the importance of self-monitoring blood glucose (SMBG) for type 1 diabetes is widely acknowledged, it is still questioned in type 2 diabetes, at last when it is not treated with insulin. Indeed, SMBG appears to be efficient only when it is integrated into a strategy of self-treatment, which leads the patient to adapt his treatment to his blood sugar tests. Passive self-control is useless, and can even favour increasing anxiety or frustration. Numerous problems are still to be solved, be it concerning the adaptation of the treatment - the number of tests per day or per week, the kind of therapeutic adaptation, etc - or concerning the therapeutic education, which should correspond to the patient's personality and to his strategy of treatment. Evaluating the locus of control of the patients could help to single out those who could benefit from SMBG. The development of SMBG, whose cost is financed by social Security in France, is a good mirror of the patients' increasing wish to be informed about their treatment and to take an active part in it. Yet prospective randomised studies are still needed to confirm the efficiency of SMBG in type 2 diabetes. PMID- 12746627 TI - [Blood glucose monitoring in the management of type 1 diabetes in childhood]. AB - In the last twenty years, blood glucose self-monitoring has attained a nearly optimal level, with reliability, easiness, quickness and safety, which makes it tolerable to a greater number of patients and allows young diabetics to make more easily something which is mandatory to the management of type 1 diabetes. The principles of treatment are identical in children, adolescents and adults. However, many patients experience difficulties in applying the ideal therapeutic model. This must be particularly taken into account in children, who are in a stage of full intellectual and psychological development. Inappropriate demands can drive them to failure and go against the main objective at this period of life, which is education. Reference documents help pediatricians to the daily practice of home blood glucose monitoring and, more generally, management of childhood diabetes: ISPAD Consensus Guidelines (ISPAD=International Society for Pediatric and Adolescent Diabetes), which has been translated in French, and Cahiers de l'AJD (AJD Note books), for educating new patients. Blood glucose monitoring is recommended: to evaluate the glycemic response to the effect of insulin; to confirm hypoglycemia; to prevent hyperglycemia in case of disease; in case of physical exercise. The frequency of home blood glucose monitoring should be individualized, depending on the acceptability for the young patients and the insulin therapeutic regimen. PMID- 12746628 TI - [Insulin therapy by insulin pump: continuous or conventional self-blood glucose monitoring? ]. AB - Use of portable pumps is increasing to achieve intensive insulin treatment. Beside unpredictable insulin absorption and insufficient reactivity to changes of insulin flow rate due to the subcutaneous route, this therapy is however limited by the lack of information on blood glucose level provided by self-blood glucose monitoring. Thus, patient interpretation only leads to speculative adaptation of pump flow rate. The lack of alert toward the risk of hypoglycemia or hyperglycemic ketogenic deviations can lead to the occurrence of deleterious metabolic distorsions, among which severe hypoglycemia stands in first rank. Starting experiences of continuous recording of interstitial glucose level by portable systems using glucose-oxidase allow on short time durations the identification of daily periods of poor metabolic control. Retrospective availability of information gives the possibility of more adequate treatment adaptations than conventional capillary blood glucose monitoring, but does not allow immediate prevention of metabolic events. Only devices providing real time or near-real time information to the patient can fulfill this function. However, the absence of tight parallelism between variations of interstitial and blood glucose levels may lead to erroneous decisions. A true continuous real time information on blood glucose level on long-term seems only expectable from implantable glucose sensors. Still under investigation, these systems should be able to insure vigilance toward the risk of hypo- and hyperglycemia on weekly or monthly periods. Initially used as complementary to capillary self-monitoring, their reliability should allow their use as substitutes for conventional monitoring, except for measurements aimed at signal calibration. Pump control by the sensor signal is conceivable if it corresponds to a direct, continuous, real time measurement of blood glucose, and subject to a simultaneous improvement of insulin infusion modes. PMID- 12746629 TI - The challenge of poorly controlled diabetes mellitus. AB - Diabetes is a growing healthcare challenge worldwide, with significant socioeconomic implications in industrialised and developing nations. Epidemiological studies indicate that diabetes is likely to reach epidemic proportions within the next few decades. A considerable proportion of people either have impaired glucose tolerance with a significant risk of development of diabetes, or have undiagnosed Type 2 diabetes. Many are poorly controlled on existing therapies, with significant implications for patients' quality of life and for healthcare expenditure. Pivotal to reducing the risk of morbidity and the development of complications and mortality is the normalisation of both fasting and postprandial blood glucose levels. Various healthcare initiatives address the attainment of this treatment goal; however, there is still a need for better disease management in both Type 1 and Type 2 diabetes. PMID- 12746630 TI - Looking for an insulin pill? Use the BRET methodology! AB - Insulin exerts its biological effects through a plasma membrane receptor that possesses a tyrosine-kinase activity. This tyrosine-kinase activity depends on the autophosphorylation of the receptor on tyrosine residues and on its dephosphorylation by protein tyrosine-phosphatases. The discovery of pharmacological agents that specifically stimulate the autophosphorylation of the insulin receptor or inhibit its dephosphorylation will be of great importance for the treatment of insulin resistant or insulin deficient patients. Bioluminescence Resonance Energy Transfer (BRET) has developed in recent years as a new technique to study protein-protein interactions. In the BRET technique, one partner is fused to Renilla luciferase, whereas the other partner is fused to a fluorescent protein (e.g. YFP, Yellow Fluorescent Protein). The luciferase is excited by addition of its substrate, coelenterazine. If the two partners interact, resonance energy transfer occurs between the luciferase and the YFP, and a fluorescent signal, emitted by the YFP, can be detected. Our work indicates that this methodology could be an important tool for the search of molecules that activate insulin receptor autophosphorylation or that inhibit its dephosphorylation. Indeed, we first showed that the activation of the insulin receptor by different ligands can be monitored using a chimeric receptor with one B-subunit fused to Renilla luciferase and the other B-subunit fused to YFP. The conformational changes induced by different ligands could be detected as an energy transfer (BRET signal) between the luciferase and the YFP, that reflects the activation state of the receptor. This methodology allows for rapid analysis of the effects of agonists on insulin receptor activity and may therefore be used in high-throughput screening for the discovery of molecules with insulin-like properties. More recently, we demonstrated that the BRET methodology could also be used to monitor the interaction of the insulin receptor with protein tyrosine phosphatase 1B, one of the main tyrosine-phosphatase that controls its activity. HEK cells were co-transfected with the insulin receptor fused to Renilla luciferase and a substrate-trapping mutant of PTP1B (PTP1B-D181A) fused to YFP. Insulin-induced BRET signal could be followed in real time for more than 30 min. Therefore, this methodology can also be used in high-throughput screening for the search of molecules that will specifically disrupt the interaction between the insulin receptor and PTP1B. PMID- 12746631 TI - Association study of the vitamin D: 1alpha-hydroxylase (CYP1alpha) gene and type 2 diabetes mellitus in a Polish population. AB - OBJECTIVES: Type 2 diabetes mellitus (T2DM) is a complex disease. Genetic and environmental factors cooperate together to form its clinical picture. Polymorphisms in genes involved in the metabolism of vitamin D may influence susceptibility to T2DM. One of them is the vitamin D 1alpha-hydroxylase (CYP1alpha) gene. In this study we searched for the association of two markers, one in its intron 6 and the another one located upstream from the 5' end of CYP1alpha gene, with T2DM in a Polish population. METHODS: Overall 522 individuals were included in this study: 291 T2DM patients and 231 controls. The sequences, which contain both examined variants, were amplified by polymerase chain reaction (PCR). The T-->C polymorphism in intron 6 was assessed by the dot blotting method using P(32). Genotyping of the other variant in the 5' end of CYP1alpha gene was carried out by restriction fragment length polymorphism (RFLP) method. Since variants of both SNPs were in very strong linkage disequilibrium, haplotypes could be assigned to phase-unknown individuals. The distribution of alleles, genotypes, haplotypes and haplotype combinations was compared between the groups by chi(2) test. RESULTS: The frequency of T/C alleles of the 5'end variant was 81.7%/18.3% in T2DM patients and 82.8%/17.2% in the controls (chi(2)=0.2, 1.d.f., p=0.65). For a T-->C polymorphism in intron 6 the frequency of alleles was 65.1%/34.9% and 67.5%/32.5% in T2DM patients and controls, respectively (chi(2)=0.413, 1.d.f., p=0.669). Distribution of genotypes, haplotypes and haplotype combinations were similar in both groups. In stratified analysis, we observed that the T-C/T-T heterozygous haplotype combination was more prevalent in the subgroup of obese T2DM patients (BMI >=30) than in the controls (41.5% vs 28.6%, p=0.01). CONCLUSIONS: Vitamin D 1alpha-hydroxylase is not a major gene for T2DM in a Polish population. However, this gene may be associated with T2DM in subjects with obesity. Thus, to definitely determine the role of this gene in T2DM further studies are necessary in other populations using larger sample size. PMID- 12746632 TI - Effect of metformin on insulin sensitivity and insulin secretion in female obese patients with normal glucose tolerance. AB - OBJECTIVES: Metformin is recognized as the treatment of chronic obese, insulin resistant type 2 diabetic patients. Whether it improves insulin sensitivity in obese patients with normal glucose tolerance remains unknown. METHODS: Eight obese female patients with normal glucose tolerance were studied during a double blinded, randomized cross-over study including a 2-week administration of metformin and a 2-week administration of placebo. Insulin secretion and insulin sensitivity were assessed after metformin and placebo by means of a 3-hour hyperglycemic clamp. RESULTS: The plasma insulin and C-peptide concentrations during the hyperglycemic clamp were identical after placebo or metformin (both first and second phases). Insulin-mediated glucose disposal, stimulation of glucose oxidation and suppression of endogenous glucose production were identical after metformin and placebo. CONCLUSIONS: Metformin does not improve insulin sensitivity nor insulin secretion in obese female patients with normal glucose tolerance. PMID- 12746634 TI - Comparison of two diagnostic tests for gestational diabetes in predicting macrosomia. AB - OBJECTIVES: To validate a diagnostic test for gestational diabetes which predicts the risk of macrosomia. METHODS: A prospective study was carried out among 354 women at risk for gestational diabetes to compare two glucose tests diagnosing pregnancies at risk of macrosomia. The "practical" test consisted in glucose measurement in the fasting state and two hours after an usual breakfast and the "reference" test was the test proposed in France (O'Sullivan test with or without a 100 g oral glucose tolerance test). Both tests were made between the 24(th) and 28(th) week of gestation. Women at high risk for macrosomia were treated. The first assessment criterion was macrosomia (babies large for gestational age). Because of the presence of women treated for gestational diabetes in our sample, the sensitivity and specificity of the tests in diagnosing pregnancies at risk of macrosomia were calculated using either the incidence of macrosomia observed in our population, or the incidence of macrosomia observed theoretically in the absence of treatment (22% in literature). RESULTS: Macrosomia was diagnosed in 49 neonates (14%). The "practical" test was significantly more sensitive than the reference test (respectively 46.9% versus 16.3%, p=0.0001 in the first case, and 54.3% versus 20.1%, p=0.0001 in the second case). The "reference" test was significantly more specific than the "practical" test (respectively 80% versus 68.2%, p=0.0001 in the first case, and 80.6% versus 70%, p=0.0001 in the second case). CONCLUSION: Our study shows that the simplified "practical" test is more sensitive than the "reference" test currently used in France in screening women at risk of macrosomia. PMID- 12746633 TI - Interpretation of circulating C-reactive protein levels in adults: body mass index and gender are a must. AB - OBJECTIVE: The recently demonstrated association between C-reactive protein (CRP) level and body mass index (BMI) raised the question of the link between CRP and the degree of obesity. In the present study, we measured CRP in a healthy population with a wide range of BMI in order to appreciate the influence of overweight in the interpretation of CRP results in clinical use. METHOD: Blood donors, aged from 19 to 65 years, were included in the study. According to BMI, subjects were classified into 3 groups: A (BMI<25 kg/m(2), n=611); B (25-30, n=147); C (> 30, n=34). RESULTS: CRP values were different among women and men. CRP progressively increased with BMI in women. These results clearly showed that average level of CRP was quite different according to BMI and gender of the subjects and generated different normal ranges of CRP expressed in mg/L (median, 75(th) percentile): Group A: women: 0.44, 0.93; men: 0.40, 0.79, Group B: women: 1.28, 1.84; men: 0.84, 2.17, Group C: women: 3.61, 7.21; men: 1.16, 3.08. CONCLUSION: Our results suggest that for an inflammatory disease diagnosis, a CRP concentration of 5 mg/L is normal for obese women but is five times the 75(th) percentile for normal people. PMID- 12746635 TI - Protection of insulin-secreting INS-1 cells against oxidative stress through adenoviral-mediated glutathione peroxidase overexpression. AB - OBJECTIVES: A large fraction of an islet graft can be lost early following allotransplantation from various non specific mechanisms including oxidative stress. Overexpression of antioxidant enzymes could confer a beneficial effect on islets exposed to reactive oxygen and nitrogen species. We examined the viability of beta cells driven to overexpress glutathione peroxidase (GPx) and exposed to a superoxide donor (hypoxanthine/xanthine oxidase HX/XO) and a nitric oxide donor (3-morpholinosydnonimine SIN-1). METHODS: Cultured INS-1 rat-derived insulin secreting cells were transfected by an E1-deleted adenovirus carrying GPx cDNA (AdGPx). Additional experiments were performed with an adenovector carrying Cu/Zn superoxide dismutase cDNA (AdSOD). Cellular viability was tested by the WST-1 colorimetric assay and functionality by static incubation. RESULTS: AdGPx increased GPx activity within 48 hours from 0 (untransfected cells) to 60 +/- 11 U/g (cells transfected at an MOI of 25: 1). GPx overexpression significantly reduced cytotoxicity induced by HX/XO from 10.81 +/- 1.41 to 5.42 +/- 2.62% at 10 mU/ml and from 61.19 +/- 4.17 to 52.9 +/- 4.39% at 20 mU/ml (p=0.0002, transfected cells vs control cells). Doses of SIN-1 from 600 to 1000 micromol/l resulted in cytotoxicity ranging from 17.66 +/- 3.48 to 45.97 +/- 6.48% in control cells and from 5.65 +/- 1.37 to 35.80 +/- 5.59% in AdGPx transfected cells (p=0.015). The combination of AdGPx and AdSOD did not exhibit any synergistic cytoprotective effect. Control cells exposed to a HX/XO stress exhibited a reduction in glucose-theophylline stimulated insulin secretion by half, while stressed GPx overexpressing-cells maintained the same insulin secretion level than non-stressed cells. CONCLUSIONS: Adenoviral-induced overexpression of GPx enhances the resistance of a rat beta cell line to both reactive oxygen (ROS) and reactive nitrogen species (RNS) cytotoxicity. Transposition of these findings to human islet transplantation with a clinically relevant procedure deserves further investigations. PMID- 12746636 TI - Control of diabetes and cardiovascular risk factors in patients with type 2 diabetes: a nationwide French survey. AB - OBJECTIVES: To evaluate in France in 2001 the therapeutic management and control of diabetes and of modifiable cardiovascular risk factors in patients with type 2 diabetes receiving specialist care. METHODS: The study was proposed to 575 diabetologists across France. The first 8 consecutive ambulatory patients with type 2 diabetes treated by oral antidiabetic drugs (OADs) and/or insulin attending for consultation with a diabetologist were eligible for inclusion in the survey. The following data were collected: demographics, diabetic and cardiovascular history, cardiovascular risk factors, blood pressure, last recorded measurements of HbA(1c) and LDL cholesterol, and details of diabetes medication and cardiovascular medication. RESULTS: 4, 930 patients (53% men) aged 62 +/- 11 years were recruited by 410 specialists in diabetes care. The mean duration of diabetes was 12 +/- 9 years. 71% of patients were treated with OADs, 18% with an OAD + insulin and 9% with insulin alone. Mean HbA(1c) was 7.6 +/- 1.6%; HbA(1c) was<=6.5% in 27% of patients, between 6.6% and 8% in 39% of patients, and > 8% in 34% of patients. Mean blood pressure was 140 +/- 16/80 +/- 9 mmHg. In the study population as a whole the target blood pressure (systolic BP<140 mmHg and diastolic BP<80 mmHg) was attained by 29% of patients. Among the 3, 085 patients (63%) treated for hypertension, this target was attained in only 23% of patients; 40% of patients treated for hypertension received one single antihypertensive treatment, 36% received 2 treatments and 24% received 3 treatments or more. Among the 1, 845 patients considered by the investigators as not having hypertension, the target blood pressure was attained by 39%. A measurement for LDL cholesterol was available in 4, 036 patients (82%). 58% of these patients had LDL cholesterol<1.3 g/l, 29% had values between 1.3 and 1.6 g/l, and 13% had values > 1.6 g/l. 52% of patients were not receiving any lipid lowering agents, 28% were treated with statins, 19% with fibrates, and 1% with statins + fibrates. LDL cholesterol was<1.3 g/l in only 66% of the 646 patients with associated coronary heart disease. CONCLUSION: According to this large nationwide survey, the prevalence of cardiovascular risk factors remains high. Control of glycaemia, blood pressure and LDL cholesterol does not appear to be optimal. This is due in part to the severity of diabetes in these patients seen by specialists in diabetes care; however, both awareness and application of published recommendations need to be reinforced. PMID- 12746637 TI - Accuracy of the continuous glucose monitoring system in inpatient and outpatient conditions. AB - OBJECTIVE: To evaluate in inpatient and outpatient conditions, using only non calibration data, the accuracy of the Continuous Glucose Monitoring System (CG MS). RESEARCH DESIGN AND METHODS: Twelve, 21 and 20 type 1 diabetic patients participated in 3- day inhospital, 6-day ambulatory (i.e., 2 sensors worn consecutively) and 3-day ambulatory studies. Capillary glucose tests (7-8/day) served as comparisons. Pairs of data (excluding the 4 daily pairs used automatically by the CGMS for calibrations) were analyzed using correlation coefficient, error -grid analysis, and mean of absolute differences. RESULTS: Two third and half of the sensing days were qualified as "optimal", i.e., interpretable by the CGMS program in the inpatient and outpatient studies, respectively. Correlation coefficient was 0.92, 0.81 and 0.73 in the inpatient, 6 day outpatient and 3-day outpatient studies. 98%, 93% and 96% of the data fell in the A or B error-grid "clinically acceptable" zones. Mean absolute differences between sensor and glucometer values were 25 +/- 2, 34 +/- 5 and 32 +/- 5 mg/dl. CONCLUSION: CGMS may be used for assessment of glycemic trends, though accuracy is more in the range of glucometers than of laboratory tests. Inhospital utilisation improved accuracy and 6-day rather than 3-day sessions may be more appropriate to evaluate day to day reproducibility of glucose profile. PMID- 12746639 TI - Marital status and family size of type 1 diabetic patients in a French cohort. AB - OBJECTIVE: To compare the marital status, the number of offspring and the cumulative incidence of type 1 diabetes in offspring of type 1 diabetic men and women. METHODS: From the database of patients attending our department, we reviewed the files of all the 352 subjects aged >=40 years with type 1 diabetes and compared male and female patients for whom age, age at diagnosis of diabetes, marital status, socio-economic status, number and age of offspring, diagnosed type 1 diabetes in the offspring could be obtained from patient's record and/or direct interview (86 males and 78 females). RESULTS: In this population, 73% of women and 81% of men were married or living a marital life (NS), and 35% of women versus 8% of men had no offspring (P<0.0001). The proportion of parents with 2 offspring or more was 43% in females and 61% in males (p=0.03) and was not related to the socio-economic status. The number of offspring with diagnosed type 1 diabetes was small (8/229) and did not show significant association with gender of the parent, with a cumulative incidence of 3.2 and 3.7% in offspring of type 1 diabetic mothers and fathers respectively. CONCLUSION: Type 1 diabetic women born before 1960 had fewer children than men. In this cohort, there was no difference in the cumulative incidence of type 1 diabetes in offspring of type 1 diabetic men and women despite reduced family size in women. PMID- 12746638 TI - Mild acute renal failure potentiates metformin accumulation in the diabetic rat kidney without further impairment of renal function. AB - OBJECTIVES: To analyze, in acute renal failure (ARF) in diabetic rats, how moderate functional ARF would modify metformin (MET) pharmacokinetics and if plasma and renal tissue MET accumulation could aggravate renal insufficiency and/or elicit plasma lactate accumulation. METHODS: Streptozotocin-induced diabetic rats were allocated to four groups: control, MET, ARF, ARF-MET (6-7 rats per group). MET (100 mg/kg/day) was given per os for two weeks before ARF was induced by drinking restriction and enalapril treatment. The effects of MET and/or ARF were examined in vivo on renal function in conscious rats (metabolic cages) and ex vivo on renal vascular reactivity (isolated kidney). RESULTS: MET treatment (plasma level: 5.3 +/- 1.4 microg/ml, mean+/-SEM), resulted in biguanide accumulation in cortex and medulla (53 +/- 17 and 80 +/- 40 microg/g respectively). MET was devoid of any effect on creatinine clearance, mean blood pressure or renal vascular resistance, but moderately increased plasma lactate (3.8 +/- 0.5 vs 3.2 +/- 0.2 mM, P<0.05) and decreased angiotensin II-induced renal vasoconstriction. ARF, although mild, decreased renal MET clearance (0.29 +/- 0.05 vs 1.01 +/- 0.31 ml/min/100 g, P<0.05) and increased plasma and renal tissue MET levels (x 2-4). MET however did not worsen the fall in glomerular filtration rate, nor modify renal vascular reactivity. ARF did not change the MET elicited moderate increase in plasma lactate. CONCLUSION: Despite the increase in MET plasma and renal tissue levels subsequent to moderate ARF, no harmful metabolic effect on plasma lactate and no further impairment of renal function was observed in MET-treated diabetic rats subjected to ARF. PMID- 12746640 TI - Obesity or diabetes: what is worse for the mother and for the baby? AB - OBJECTIVES: The aim of the present study is to evaluate pregnancy outcomes in a cohort of Caucasian pregnant women in relation to their body mass index and glucose tolerance status; the role of central fat distribution, as indicated by waist-to-hip circumference ratio, was also considered. METHODS: Seven hundred women were studied; they had gestational diabetes or impaired glucose tolerance (250) or normoglycaemia (450). Among them 117 had pre-pregnancy overweight/obesity (44 were obese), 133 hyperglycaemia, but normal weight, and 117 hyperglycaemia and overweight/obesity (42 were obese). RESULTS: Hypertension, cesarean delivery and prevalence of large-for-gestational age babies were higher in obese (both with normoglycaemia and hyperglycaemia), mainly in those with greater gestational weight gain and central fat distribution (waist-to-hip ratio > 0.90). Normal weight hyperglycaemic women showed better outcomes than obese normoglycaemic women did. In a multiple logistic regression model, obesity (OR=10.6; 95% CI 5.00-22.54) was directly related to hypertension, and independent predictors of cesarean section were: gestational hyperglycaemia (OR=1.78; 95% CI 1.21-2.62), gestational weight gain (OR=1.06; 95% CI 1.02-1.10), and central obesity (OR=1.51; 95% CI 1.02-2.24), while obesity (OR=4.48; 95% CI 2.30-8.71) gestational weight gain (OR=1.08; 95% CI 1.03-1.12) and central fat distribution (OR=1.81: 95% CI 1.12-2.93) were directly related to delivering larger babies, after multiple adjustments. CONCLUSION: These results suggest that pre-pregnancy obesity and gestational hyperglycaemia were independent risk factors for different adverse pregnancy and neonatal outcomes, while central distribution of fat, and gestational weight gain play an additive adverse role on these outcomes. PMID- 12746641 TI - [Asymptomatic glycemic instability: how to measure it and which clinical applications? ]. AB - The glycemic variations observed in nondiabetic subjects are primarily related to the post-prandial metabolic responses generated by the food intake. This physiological glycemic variability is reproducible between two successive days under conditions of similar meals and activities. In insulin-treated diabetic patients, glycemic instability is directly related to loss of glucose homeostasis due to the failure then the deficiency of endogenous insulin secretion, but its physiopathological implication in term of diabetic angiopathy risk remains to be determined. Other exogenous glycemic instability factors are known and have to be corrected (not adapted repartition of the meals and/or the insulin doses during the day, inappropriate choices in insulin regimens, errors in insulin adjustments, inadequate insulin injection). Classical glycemic measurements used to appreciate the quality of the glycemic control do not provide or give partial and erroneous information on quality of glycemic stability. It's particularly the case for glycated hemoglobin and glycemic average. In order to exactly quantify glycemic variability, specific tools of measurement or calculation can evaluate the intra-day glycemic variations or excursions (MAGE), the day-to-day glycemic variations or excursions (MODD), the meal-related glycemic excursions (MIME) or the risk of severe hypoglycemia (LBGI). Some of these tools are easy to use, other much complex or difficult to apply in clinical practice, even by using new methods such as the continuous glucose monitoring system (CGMS). Thus, asymptomatic glycemic variability is related to loss of glucose homeostasis, but also to irrelevant or incompatibility of the treatments used to obtain glucose control in insulin-treated diabetic patients. The evaluation of glucose instability can be provided using new methods of glucose monitoring and could be useful to optimize the therapeutic regimens in some of these diabetic patients. PMID- 12746642 TI - Crohn's disease plus Takayasu's arteritis: more than coincidence? PMID- 12746643 TI - [Respiratory syncytial infections in the elderly. Seven cases and review of the literature]. AB - We report 7 cases of respiratory syncytial virus (RSV) infections among elderly patients hospitalized for acute lower tract infection during the winter months. The median age was 95 years (range: 79-106 years). 6 patients were living in nursing homes. All patients had chronic cardiopulmonary conditions. Clinical symptoms included upper respiratory tract symptoms for 3 of them and lower respiratory tract symptoms for all of them. Three patients developed a severe infection with acute respiratory failure. The chest X ray showed an interstitial infiltrate in 3 cases, a consolidation in one case. All patients received oxygen, respiratory physical therapy, nebulized B-agonists and antibiotics. Corticosteroids were used for 4 patients. Progress was favorable for all patients and none died. Diagnosis was obtained in a few hours by direct antigen detection by enzyme immunoassays (EIA) in sputum. Respiratory syncytial virus infection is a common cause of respiratory tract infection in the elderly, during the winter months. Clinical manifestations are similar to influenza infections. Detection of RSV antigens from sputum specimens may be a useful and rapid diagnostic method and merits further evaluation in the elderly. Early diagnosis of respiratory syncytial virus infection should be encouraged to allow the implementation of effective infection control procedures and avoid inadequate therapies. PMID- 12746644 TI - Association of Takayasu's arteritis and Crohn's disease. Results of a study on 44 Takayasu patients and review of the literature. AB - Takayasu's arteritis and Crohn's colitis are exceptionally associated and characteristics of patients affected with both diseases have not been fully described. In a group of 44 consecutive Takayasu patients, 4 also had Crohn's disease. This 9% prevalence (95% confidence interval=2.5-21.7%) of Crohn's disease in the group is significantly greater than the highest reported prevalence of the disease in the general population. In these 4 patients, fulfilling 5 or more ACR criteria for Takayasu's disease, Crohn's disease was confirmed with colonoscopic examination and biopsies. Takayasu-Crohn patients were younger at diagnosis and tended to have systemic symptoms more frequently. Other clinical, radiological and biological characteristics were not different between the 2 groups. In the literature, 16 cases of this unusual association have been reported. The diagnosis of Takayasu's disease was simultaneous or posterior to Crohn's disease in 87%. Data on vascular lesions was available in 6 out of 16 patients and in the 4 patients of the present study: the distribution of vascular lesions in Takayasu-Crohn patients did not appear to be different from Takayasu patients. Our results and previous reports suggest that this association of rare diseases is not fortuitous. With respect to the indolent course of Takayasu's arteritis, early diagnosis of vascular lesions in Crohn patients is encouraged. PMID- 12746645 TI - [Early response to oral cobalamin therapy in older patients with vitamin B12 deficiency]. AB - PURPOSE: Standard treatment of vitamin B12 deficiency involves regular intramuscular cobalamin administration. The aim of this study was to determine whether oral cobalamin treatment may be an effective therapy for treating older patients with cobalamin deficiency related to nutritional deficiency and food cobalamin malabsorption. PATIENTS AND METHODS: We prospectively studied 20 patients older than 80 years with established cobalamin deficiency related to food-cobalamin malabsorption (n=14) and nutritional deficiency (n=6) who received 1000 micro g of oral cyanocobalamin per day. Levels of serum cobalamin and blood counts were determined at baseline and after the first week of treatment. RESULTS: After an average of 8 days of treatment, 17 out of 20 patients normalized their serum cobalamin levels; the patients had increased their serum cobalamin level (mean increase of 0.23 micro g/L; p<0.01 compared with baseline), reticulocyte count (mean increase of 27400/mm(3); p<0.05), hemoglobin levels (mean increase of 0.7 g/dL; NS), and decreased the mean erythrocyte volume (mean decrease of 0.7 fL; NS). CONCLUSION: Our findings suggest that cyanocobalamin given orally during one week may be an effective treatment for cobalamin deficiency related to food-cobalamin malabsorption and nutritional deficiency and may avoid painful intra-muscular injections in older patients. PMID- 12746646 TI - [Superantigens and vasculitis]. AB - Superantigens are microbial products, that may activate in a polyclonal way, a high number of T lymphocytes bearing a specific VB segment of TCR; the best characterized are staphylococcal and streptogenic toxins. In a review of the literature, we summarize the role of these superantigens in primitive vasculitis, specially in Kawasaki syndrome and Wegener's granulomatosis. This pathogenic model highlights the complex relations between infection and autoimmunity. PMID- 12746647 TI - [Bobigny: the Fifth Franco-American Exchange Meeting in Endocrinology: relationships between hormones and cancers--patient education]. PMID- 12746648 TI - [SERMs and uterus]. AB - The uterus is one of the target organs of sexual steroids synthesized in the ovary. Estrogen is known to stimulate cell proliferation in the endometrium while progesterone has an anti-estrogenic secretory effect on this tissue. Renewed interest in the action of new anti-estrogenic agents on the uterus has arisen over the last decade, but not simply in order to achieve new therapeutic strategies for the prevention or cure of uterine tumors. New compounds were developed for their action on other tissues such as the breast, but it rapidly became clear that they were a source of uterine disease. A clear example is tamoxifen which has a powerful anti-estrogenic effect on breast tissue. It was hoped however that this compound, which behaves either like an antagonist or an agonist, depending on the target tissue, could have an anti-estrogenic effect on the uterus and on the contrary an agonistic estrogenic protective effect on bony and vascular tissue. This approach progressively led to the development of SERMs (Selective Estrogen Receptor Modulators), non-steroidal compounds modulating the action of estrogens. The first member of this new pharmaceutical class was raloxifen, marketed in France under the brand name Evista, which has an estrogenic effect on vertebral bone, warranting its authorization for use in patients with vertebral osteoporosis with or without fracture. Raloxifen thus has a beneficial estrogenic effect, at least on trabecular bone, and an anti estrogenic effect on the uterus and breast. The goal today is to continue the development of new compounds in the SERM family with well targeted, and well understood, agonistic and/or antagonistic actions on different body tissues. PMID- 12746649 TI - Medullary thyroid carcinoma and hormones. AB - Medullary thyroid carcinoma (MTC) is a neuroendocrine tumor. The identification of calcitonin as a specific and sensitive marker in 1968 determined a special interest of endocrinologists. Many hormones have been isolated in MTC tumors. Calcitonin, prostaglandins and somatostatin are of particular interest. Calcitonin is not only a marker, but is indicative of tumor size and prognosis. Moreover the presence of calcitonin receptors in MTC could have a role in the proliferation of this tumor. Somatostatin and the large use of somatostatin analogs in the treatment of neuroendocrine tumors raise the hope that these drugs could be effective in MTC patients. Unfortunately, the benefits of these treatments is not validated. More interestingly, nonsteroidal anti-inflammatory drugs could be of interest in this tumor by comparison of their use in colorectal polyps. By inhibiting Cox or inducing 15 PGDH enzymes, we have demonstrated in vitro its effectiveness. Thus, hormones in MTC are not only markers but probably mediate biological effects involved in tumor growth. PMID- 12746650 TI - Observance in diabetes: from therapeutic education to therapeutic alliance. AB - The aim of this paper is to understand why non-observance is so frequent in diabetic patients, to delineate some of its mechanisms, which should make it possible to propose approaches to prevent non-observance. This will lead us to clarify the relationship between therapeutic education and observance, and to justify on a theoretical basis the concept of therapeutic alliance. PMID- 12746651 TI - [Hepatitis C and tricholeukocyte leukemia: a fortuitous association? A case report]. AB - The association of hepatitis C and certain hemotological diseases (non-Hodgkin B cell lymphoma, villous splenic lymphoma.) is a debated question which remains open due to discordant epidemiological data. We report the case of a new patient with chronic hepatitis C and tricholeukocyte leukemia. PMID- 12746652 TI - [Kikuchi-Fujimoto disease: two cases]. AB - Kikuchi-Fujimoto disease, or necrotizing histiocyte lymphadenitis, is a rare anatomo-clinical entity of unknown cause. The patients present painful cervical lymph nodes, fever, and a flu-like syndrome, but the spontaneous course is usually favorable in six months. We observed two cases within six months in our unit. Both patients improved rapidly after diagnostic adenectomy. PMID- 12746653 TI - [Psoriasis and systemic lupus erythematosus: a rare association with specific therapeutic problems]. AB - The association psoriasis and systemic lupus erythematosus (SLE) is a very uncommon association. We report three cases, diagnosed in an Internal Medicine department between 1993 and 2000. Few cases of psoriasis/SLE have been published in the literature. Psoriasis generally precedes the diagnosis of SLE. Psoriasis can also be associated with discoid lupus erythematosus. In some cases, SLE appears as a complication of ultraviolet phototherapy indicated for the psoriasis. The association psoriasis/SLE does not seem to have distinctive immunologic features. Specific therapeutic difficulties may occur. Indeed, hydroxychloroquine may exacerbate the psoriasis. Systemic use of corticosteroids raises the risk of severe psoriasis relapse during withdrawal. In addition, the diagnosis of psoriasic arthropathy is more difficult in this setting. The psoriasis/SLE association might be a good indication for using methotrexate. PMID- 12746654 TI - [Primary biliary cirrhosis and autoimmune hepatitis overlap syndrome: therapeutic features in 5 patients]. AB - Autoimmune hepatitis (AIH) and primary biliary cirrhosis (PBC) are two different liver diseases, however diagnosis criteria of these two affections can be found in a same patient. The aim of this study is to relate the clinical, serologic and histologic features of patients presenting the overlap syndrome, AIH-PBC, defined by the presence of at least of two main criteria of each disease and to evaluate their response to therapy. PATIENTS AND METHODS: This was a retrospective study concerning patients presenting overlap syndrome diagnosed between January 1998 and December 2001. These patients had been treated with ursodesoxycholic acid (AUDC) or prednisone and azathioprine or a combination of these three products. Clinical and biological criteria were used to assess response to therapy. RESULTS: Five patients fulfilled the diagnostic criteria of overlap syndrome. All patients were females, the median age was 38 years (range: 19-65 years). Three patients were treated by UDCA, a clinical and biological response was noted in only one patient. Two patients were treated by prednisone and azathioprine without any remission. Three patients were treated by a combination of these three products with a good response in two cases. CONCLUSION: Optimal treatment for overlap AIH-PBC syndrome remains to be determined. Treatment with UDCA or immunosuppressor alone is not efficient. A combination of these drugs should be evaluated in further studies. PMID- 12746655 TI - [Progress and stagnation in chemotherapy protocols for primary osteosarcoma]. AB - Thirty years ago, osteosarcoma of a limb meant amputation and death. Two years after diagnosis, 80% of the patients died from pulmonary metastases, despite early amputation. In 2002, more than 70% of these patients can be cured. Successful cure requires adequate and early management centered on en-bloc resection by a specialized surgeon, followed by a long, expensive, and adapted chemotherapy. The cornerstone is high-dose methotrexate. We present data accumulated over the last 30 years on the treatment of osteosarcoma and point out the fundamental steps of this success story. PMID- 12746656 TI - [Role of the nucleosome in the physiopathology of systemic lupus erythematosus]. AB - Evidence is to date accumulating to suggest that the nucleosome, the fundamental unit of chromatin and ubiquitous product of cell apoptosis, plays a key role in the pathogeny of systemic lupus erythematosus (SLE). Nucleosomes play a central role in the antinuclear antibody response in SLE. Lupus anti-dsDNA and antihistone antibodies are directed towards nucleosomes and belong together with nucleosome-specific antibodies to a broad anti-nucleosome antibody family. Besides anti-dsDNA, nucleosome-specific antibodies have a major role in the pathophysiology of SLE and emphazise the role of nucleosome-antinucleosome immune complexes. Antinucleosome IgG antibodies are a more sensitive marker of SLE than anti-dsDNA. High levels of antinucleosome IgG are almost exclusively found in SLE. Antinucleosome IgG3 are strongly correlated with the SLE disease activity index. Nucleosome can bind to the surface of several cell types and mediate the binding of antinucleosome antibodies. Understanding of the key role of the nucleosome has opened new therapeutic intervention in SLE, such a tolerance induction to the subnucleosomal particles. PMID- 12746657 TI - [Classification of dermatologic manifestations in lupus erythematosus]. AB - Various dermatologic manifestations are observed in the different lupus subsets. Lupus lesions are characterized by a dermo-epidermal dermatitis. Other lesions, vascular or non vascular, are essentially present in association with systemic lupus erythematosus. Acute, subacute and chronic lupus erythematosus are distinguishable according to their clinical aspects, pathological features and evolution. Acute lesions are either localized to the midface or widespread. Subacute lesions may be annular or psoriasiform. Chronic lupus erythematosus includes localized or widespread discoid lupus, lupus tumidus, chilblain lupus and panniculitis. Therapy of cutaneous lupus is mainly based on antimalarials and avoidance of sun irradiation. In refractory cutaneous lupus, no universal guidelines are currently available. Except for acrosyndromes and urticaria-like lesions, vascular lesions may be due to vasculitis or thrombosis. An accurate diagnosis is absolutely necessary since therapy is different in thrombosis and vasculitis. Non vascular and non lupus lesions are numerous, some of them require special treatment such as dapsone for bullous lupus. PMID- 12746658 TI - [Ovulation induction therapy and systemic lupus erythematosus]. AB - Improvement in the prognosis of SLE prognosis has led to considering infertility therapy. The earliest reports displayed complications such as SLE revealed by ovulation induction or thrombophlebitis. Fertility is known to be normal in women with SLE, excepting amenorrhea accompanying severe flare-ups, renal insufficiency related hypofertility and ovarian failure secondary to cyclophosphamide therapy. Anti-phospholipid antibodies are suspected to cause defective nidation and placental ischemia. An exponential rise of serum estradiol is observed irrespective of the ovulation induction protocol used, leading to SLE flare-up and thrombosis. We have experience with 114 cycles in 21 women with SLE and/or APS. A complication (fetal loss, SLE flare-up, thrombophlebitis) revealed the underlying disease in 8 women. Eighteen pregnancies led to 9 live-births, 4 fetal deaths and 5 embryonic losses. Pregnancy rate was higher after ovulation induction using gonadotropins (25% per cycle), than clomiphene (4%). Pregnancy rate was similar after IVFETE, whether the protocol was planned or not. However, three-quarters of the pregnancies after unplanned IVFETE led to abortions. On the contrary, 6 out of 7 pregnancies after planned IVFETE led to live-births. Two women developed thrombophlebitis after gonadotropins therapy. A SLE flare-up appeared after 13 out of 62 cycles, with a flare-up rate higher after gonadotropins (27% per cycle) than clomiphene therapy (6%), and after an unplanned (30%) than a planned procedure (10%). In conclusion, ovulation induction therapy can reveal SLE or APS. Clomiphene complications are uncommon. When gonadotropin therapy is considered, a preventive anti-inflammatory therapy should be discussed in SLE patients, in conjunction with heparin and/or anti aggregate therapy for those with asymptomatic anti-phospholipid antibodies or prior thrombotic events. PMID- 12746660 TI - [Insulin autoimmune syndrome revealing systemic lupus erythematosus]. AB - A 22-year-old woman was admitted in August 2001 for loss of consciousness due to hypoglycemia. Her serum insulin level during the hypoglycemic episode was high at 121 mU/l (normal range: 5-25 mU/l). She had never received an insulin injection. Insulin antibodies by radioimmunoassay were positive. During hospitalisation, the patient presented clinical and biological features of systemic lupus erythematosus (SLE). Treatment with high-dose corticosteroids and cyclophosphamide resulted in restoration of euglycemia associated with resolution of circulating anti-insulin antibodies and parallel improvement in clinical and laboratory features of SLE. PMID- 12746659 TI - [Anti-phospholipid syndrome nephropathy]. AB - In the course of the antiphospholipid syndrome (APS), the renal involvement attributable to primary APS, APS nephropathy (APSN), corresponds to vaso occlusive lesions of the intrarenal vessels, associated side-by-side, acute thromboses with chronic arterial and arteriolar lesions, leading to zones of cortical ischemic atrophy. A second retrospective study of 114 lupus patients was undertaken to determine whether APSN can be found in the course of systemic lupus and if there are biologic or clinical predictive factors of the presence of APSN. We showed the following PMID- 12746661 TI - [Hypoalbuminemia and invalidating edema in an elderly subject: treatment with parenteral albumin infusion]. AB - Many factors can lead to edema. Hypoalbuminemia and the inability of the renal distal tubule to excrete salt are a common cause of edema, that can be due to a defect of albumin synthesis or malnutrition, kidney disease, protein-losing enteropathy, or increased protein catabolism. Hypoalbuminemia is frequent amongst elderly people. Its prevalence ranges from 4% to 50% depending on whether they live at home or in hospice. In normal circumstances, albumin has a double function to maintain oncotic pressure and to transport endogenous or exogenous substances. Lack of albumin is thus responsible for numerous pathological states. In this article, we report two clinical observations in patients aged over 65, suffering from generalized, highly invalidating edema, with low albumin states due to malnutrition. A rapid long-term recovery was observed after administrating albumin, and produced a far better quality of life for the patients. The clinical results obtained following this specific indication warrant further research work to verify their scientific validity on a larger scale. PMID- 12746662 TI - [Non-metastatic thrombosis of the superior sagittal sinus complicating Burkitt's lymphoma. A case report]. AB - We report a case of non-metastatic superior sagittal sinus thrombosis complicating a Burkitt's lymphoma of the small bowel, and review similar cases of this cerebrovascular disorder associated with lymphoma. PMID- 12746664 TI - [Vitiligo: nothing new?]. PMID- 12746663 TI - [Pleural effusion as a first sign of Ig D lambda multiple myeloma]. AB - Immunoglobulin D (IgD) multiple myeloma is rare, accounting for less than 2% of all patients with multiple myeloma. The main presenting features are bone pain in 70% of patients. Extramedullary involvement is less common. We report a case of Ig D lambda multiple myeloma in a 74-year-old man that was revealed by pleural effusion and dyspnea. This effusion was found to be caused by multiple myeloma after electrophoretic and cytologic assays. The patient received a course of chemotherapy with melphalan and prednisone. The patient died one month later with signs of septic shock. Pleural effusion as a first sign of Ig D multiple myeloma is rarely described and the prognosis associated with such a localisation is very poor. PMID- 12746665 TI - [Cutaneous necrosis is predictive of cancer in adult dermatomyositis]. AB - INTRODUCTION: Adult dermatomyositis is associated with cancer in 15 p. 100 to 50 p. 100 of cases and, hence, investigations should be systematically performed to search for cancer. A number of predictive factors have been reported. The aim of our study was to search for predictive factors of cancer, among adults with dermatomyositis. METHODS: We prospectively assessed 26 adults presenting with dermatomyositis, hospitalised in our department of dermatology from January 1993 to June 2000. The parameters assessed were: association with a cancer, age, gender, cutaneous necrosis, muscular weakness, electromyographic abnormalities, erythrocyte sedimentation rate, and muscular enzyme levels. RESULTS: Mean age was of 52 years and sex ratio (M/F) was of 0.53. Cancers were diagnosed in eight cases (31 p. 100) (mean age: 59.5 years; sex ratio=1; cancer localization: lung (2), breast (2), ovary, endometrium, bladder, and melanoma). Five patients in the cancer group had cutaneous necrosis and only 2 in the without cancer (p=0.01; PPV=71.4 p.100). Elevation of muscular enzyme was also associated with cancer. CONCLUSION: Our report demonstrates that cutaneous necrosis is closely associated with cancer and it suggests that in selected patients with dermatomyositis and cutaneous necrosis, more exhaustive and repeated investigations should be performed to search for cancer. The interest of elevation in muscular enzyme as a predictive factor of cancer is discussed. PMID- 12746666 TI - [Pseudoxanthoma elasticum: 11 cases]. AB - INTRODUCTION: Pseudoxanthoma elasticum is a congenital dystrophy of the connective tissue. Its clinical expression is cutaneous, ocular and cardiovascular. PATIENTS AND METHODS: The aim of our study was to specify the principle characteristics of this disease and to discuss the interest of various supplementary examinations in its diagnosis and control in a series of 11 patients. RESULTS: The study included 9 women and 2 men (sex ratio: 4.5). The mean age the onset of the symptoms was of 18 years. In 4 cases there was a family history of the disease. All the patients exhibited yellowish, pigskin, and papular lesions on the sides of the neck. The systematic ophthalmologic examination revealed angioid streaks in 7 cases and a pigskin aspect in 4 cases. The systematic cardiovascular and metabolic explorations revealed no abnormalities specific to pseudoxanthoma elasticum. Two cases of asymptomatic nephrocalcinosis were observed. DISCUSSION: In our patients, the disease was probably of autosomal recessive transmission. The predominance of women consulting for the disease would be explained by the esthetic damage. Diagnosis of pseudoxanthoma elasticum is based on clinical, histological and genetic criteria. Supplementary explorations are useful to confirm the diagnosis and also for the search to other, visceral, localizations. Such examinations vary depending on the teams, means and above all the evocative signs. PMID- 12746667 TI - [Immediate hypersensitivity is rarely implicated in drug induced urticaria]. AB - INTRODUCTION: The unexpected appearance of acute urticaria during the course of drug treatment gives rise to the following question: is it an allergic urticaria (due to an immediate hypersensitivity: IgE mediated specific immunity) or is it pseudo-allergic? We report our findings in an immuno-allergological study of patients who were sent for drug intolerance which presented as immediate hypersensivity (urticaria, angiooedema, anaphylactic shock). METHODS: A prospective study was conducted including all the patients who were sent to the unit for urticaria or angiooedema type drug intolerance. Patients were questioned about previous chronic urticaria and also about urticaria after taking different medicines. The clinical examination looked for a dermographism. All the patients then took skin tests for immediate hypersensitivity, the molecule was contra indicated and tests for cross-reactivity were conducted. PATIENTS: Three hundred fifty patients were sent to this unit between February 2000 and April 2001 for drug intolerance, mostly with urticaria/angiooedema but in 7 cases with anaphylactic shock. The incriminated drugs were varied: 50 p. 100 were due mainly to penicillins and cephalosporins. Other drug groups were also involved: non steroid anti-inflammatories, aspirin and paracetamol for the most part, along with local anesthetics, morphine-based products, contrast iodine products, corticosteroids. RESULTS: Of the 350 patients tested, only 22 were allergic and had positive tests for the incriminated drug. In these 22 patients, with the exception of 2 of them, the effects were severe (anaphylactic shock in 7 patients) and the urticaria was only a minor manifestation of the reaction. The drugs responsible were cephalosporin (10 patients), the penicillin (6 patients), insulin (2 patients), gonadorelin (1 patient), carboxymethylcellulose (1 patient), lidocain (1 patient), and sulfamethoxazole (1 patient). The 328 other patients had negative tests and were able to retake the tested molecule without incident. Most of them had antecedents of chronic urticaria or dermographism. DISCUSSION: Only 22 patients of the 350, i.e. 6 p. 100 were genuinely allergic. These patients were those who presented the most severe symptoms. The other patients, i.e. the majority, suffered from pseudo-allergic drug-induced urticaria, which made retaking the medicines possible. PMID- 12746668 TI - [Intravenous pulse methylprednisolone therapy for severe alopecia areata: an open study of 66 patients]. AB - INTRODUCTION: Treatment of alopecia areata is a difficult challenge. Some European publications have shown encouraging results with high dose pulse corticosteroid therapy in extensive plurifocal alopecia areata. We undertook a prospective open study between January 2000 and December 2001 using repeated pulse each month, with the aim of identifying the effects of this repetition and underlining the best indications. PATIENTS AND METHODS: Sixty-six patients aged 9 to 60 years old presenting an extensive alopecia areata exceeding 30% of the scalp surface (n=47), alopecia totalis (n=8), alopecia universalis (n=8), ophiasic alopecia (n=3), for less than 12 months entered this study. The administered treatment was methylprednisolone 500 mg/d during 3 days or 5 mg/kg twice per day during 3 days in children. These pulses were repeated after 4 and 8 weeks, then a second series was carried out or not according to cases. The main evaluation criterion was the percentage of new terminal hair appearing on the bald areas, appreciated by clinical and photographic evaluation at 3 and 6 months. RESULTS: Ophiasic alopecia areata did not respond to treatment. A quarter of patients presenting universal alopecia had a good response (higher than 80 p. 100) followed by a relapse in half the cases. Half of the patients presenting alopecia totalis had a good response, which was maintained three times out of four. Multifocal alopecia areata seems the best indication since the patients under study presented a good response in 63.8 p. 100 of cases (78 p. 100 when it was a first episode and 90.5 p. 100 if the treatment had been started in less than 3 months before). The repetition of the pulses did not appear to increase the number of responders. CONCLUSION: This study provides the best indication of pulse methylprednisolone therapy: first recent episode of extensive plurifocal alopecia areata. These results are less convincing in long term history or other forms of alopecia areata. PMID- 12746669 TI - [Rapp-Hodgkin's syndrome: two cases]. AB - INTRODUCTION: In 1968, Rapp and Hodgkin described a family (a mother and her son and daughter) with a combination of dysplasic hair, nails, hypodontia and hypohidrosis associated with oral clefting and hypoplasia of the maxillary. Forty other cases of this congenital disorder have since been reported, and provide better knowledge of this predominantly hereditary disease. CASE REPORT: We examined a 4 year-old boy for who had suffered since birth from a crusting and inflammatory scalp dermatitis which evolved periodically. He also had a coarse, wiry, light colored and slow growing hair, onychodysplasia, skin atrophy areas on the trunk, and distinctive craniofacial anomalies including a bifid uvula. He had never been seen to sweat. His father presented similar features, but less pronounced, and with a segmentary distribution. DISCUSSION: We report two new cases of Rapp-Hodgkin's ectodermal dysplasia. We suspect a mosaicism in the father who may be the first and sporadic case of the family. These discret forms are frequently misdiagnosed and we hope we have provided some new elements which will help in the description of this rare disease. Early diagnosis permits an early follow-up and avoids numerous complications. PMID- 12746670 TI - [Eccrine angiomatous hamartoma: an atypical case]. AB - BACKGROUND: We report a case of eccrine angiomatous hamartoma. This rare and benign skin disease is histologically characterized by the proliferation of both eccrine glands and vascular structures. CASE REPORT: A 65 year-old man had developed during the previous 8 years post-traumatic, angiomatous, extensive and plurifocal lesions of the left lower limb. There was neither pain or local hyperhidrosis. The diagnosis of eccrine angiomatous hamartoma was made on histological evidence. DISCUSSION: This observation is original because of the onset of the disease after 50 years of age, its widespread and plurifocal character and probable triggering-off by minor local trauma. The classical surgical treatment of eccrine angiomatous hamartoma was not suitable and a laser CO2 therapy was undertaken instead. The results were very discreet. PMID- 12746671 TI - [Systemic necrobiotic xanthogranuloma with initial pericardial and pulmonary involvement]. AB - INTRODUCTION: Necrobiotic xanthogranuloma is a rare disease, usually associated with a monoclonal gammapathy. We report a case with pericardial and pulmonary involvement that preceded the appearance of the cutaneous lesions. OBSERVATION: A 58 year-old woman was hospitalized for cardiac tamponade. She has a past history of a ductal carcinoma of the right breast that had been completely cured. Pericardial and pulmonary samples showed a predominantly histiocyte monomorphous infiltrate. Twelve months later, indurated papular-like cutaneous lesions appeared around the eye orbits and on the thorax. The histological examination of the skin revealed a necrobiotic xanthogranuloma and the diagnosis of cutaneous and visceral necrobiotic xanthogranuloma was retained. The patient exhibited an IgG Kappa monoclonal gammapathy of undetermined significance. Sequential treatment with melphalan and general corticosteroids moderately improved the cutaneous lesions but did not modify the monoclonal peak. DISCUSSION: To our knowledge, this is the first case of pericardial involvement of a necrobiotic xanthogranuloma. The discovery of the disease, revealed by a visceral manifestation at the onset is uncommon. The subsequent appearance of typical cutaneous lesions permits the retrospective diagnosis of systemic necrobiotic xanthogranuloma. Because its diagnosis is difficult in the absence of concomitant cutaneous involvement, visceral localizations of the disease must be recognized. PMID- 12746672 TI - [Cutaneous lymphangitic carcinomatosis and acquired ichtyosis associated with prostatic carcinoma]. AB - BACKGROUND: Prostatic adenocarcinoma is exceptionally associated with cutaneous lesions. We describe a patient with cutaneous lymphangitis and paraneoplastic ichtyosis related to prostatic cancer. CASE REPORT: A 92 year-old man had been treated for five years for a prostatic carcinoma. An angiomatous lesion developed with in 3 months near the right breast. Physical examination revealed axillary node enlargement, a large skin angiomatous lesion, and ichtyosis. The skin biopsy of the angiomatous skin lesion demonstrated its prostatic origin with carcinomatous metastases in the lymphatic vessels. The ichtyosis was considered as paraneoplastic. DISCUSSION: Cutaneous metastases from prostatic carcinoma are rare. Less than 1 p. 100 of cutaneous metastases are of prostate origin despite the high frequency of this cancer in the general population. The clinical aspects - angiomatous lesion and paraneoplastic ichtyosis - are exceptional. PMID- 12746673 TI - [Sensitive chronic inflammatory demyelinating polyradiculoneuropathy in Schnitzler's syndrome]. AB - BACKGROUND: Schnitzler's syndrome is a rare etiology of chronic urticaria. The disease is characterized by the association of chronic urticaria, intermittent chronic fever, bone pain, osteosclerotic bone lesions and IgM monoclonal gammapathy. More than fifty patients with this syndrome have been reported since Schnitzler reported the first case in 1974, but neuropathies are seen only in a few cases. CASE REPORT: Our patient developed, eight years after the diagnosis of Schnitzler'syndrome, a peripheral sensitive neuropathy. Anti-myelin-associated glycoprotein antibodies were not significant. A nerve biopsy specimen has revealed aspecific demyelinization. Direct and indirect immunofluorescence were negative. We conclused is that our patient presented chronic inflammatory demyelinating polyneuropathy. DISCUSSION: We found one published case of Schnitzler's syndrome and myelin-associated glycoprotein reactive peripheral neuropathy. The diagnosis of chronic inflammatory demyelinating polyneuropathy is a diagnosis of elimination. It is not the most common neuropathy associated with monoclonal gammapathy. To the best of our knowledge, it is the first case of Schnitzler's syndrome with this type of neuropathy. But there are some descriptions of chronic inflammatory demyelinating polyneuropathy presenting autoantibody activity against a myelin component, up to two years after the diagnosis of IgM monoclonal gammapathy. PMID- 12746674 TI - [Erythroderma and multiple cutaneous necrosis revealing a dermatomyositis]. AB - INTRODUCTION: We report an original case of dermatomyositis associated with neoplasia, which initial clinical expression was erythroderma and multiple cutaneous necrosis. OBSERVATION: A 64-year-old patient was admitted at hospital for erythroderma. He had a diffuse and inflammatory erythema with thrill, periorbital oedema, periungueal telangiectasia and epidermal necrosis. Physical examination also revealed symmetric proximal muscle weakness as well as hepatomegaly. There were biological signs of myolysis. Complementary investigations revealed a liver carcinoma with lung metastasis. The patient first underwent topical corticosteroid treatment, which provided partial improvement of the clinical and biological signs of disease. Thereafter he was treated with prednisone and tamoxifen. Death occurred at home 4 months after the diagnosis. DISCUSSION: Diagnosis of dermatomyositis was definitely set according to the criteria of Bohan and Peter. Epidermal necrosis, present in our observation, occur classically in dermatomyositis where they are a predictive factor of association with neoplasia. Other recognized predictive factors are the age of the patient and persistent itching. Erythroderma linked to dermatomyositis is a very unusual event: 4 cases have been reported in the literature, only one of which was associated with cancer. On the other hand, numerous cases of diffuse erythema were reported, which are close to erythroderma. It is not possible to set out that this clinical form is a factor of bad prognosis of dermatomyositis. As a matter of fact, in the published cases, one does not find more frequent deceases, pejorative signs or increased frequency of association with neoplasia. PMID- 12746675 TI - [Perianal abcess in infant]. AB - INTRODUCTION: Perianal abcess and fistula-in-ano are particular when happening in infants of less than 1 year of age. A congenital abnormality of crypts of Morgagni may be at the origin of the disease. There is no evidence for predisposing condition at this age. CASE REPORT: A 7-week-old boy had a painful swollen perianal lesion suggesting the presence of perianal abcess. Biological tests did not show any neutropenia nor inflammatory syndrome. The infant was admitted to the surgical center to undergo a perianal abcess drainage. During the intervention, no fistula was found. No recurrence was observed. DISCUSSION: Perianal abcess and/or fistula-in-ano are relatively common conditions in infants of less than 1 year of age. This disease is suspected to originate from anal cryptitis, which will later form a perianal abcess. Androgen excess during the foetal stage could be the cause of the formation of abnormal crypts of Morgagni, which encourages cryptitis and abcess formation. In childhood some predisposing factors can exist, as immunodeficiency or colopathy. On the opposite, a predisposing condition is rare in infancy. Twenty-eight to 85 per cent of infants with perianal abcess may progress to form a fistula. The usual treatment of perianal abcess is incision and drainage; it may be completed by fistulotomy or fistulectomy. The antibiotherapy is not systematic. The identification of the corresponding crypt may be important to avoid recurrence. On the opposite, a recent prospective study proposes a non operative management of perianal abcess and fistula-in-ano in healthy infants. PMID- 12746676 TI - [Tumor of the knee]. PMID- 12746677 TI - [Ulcers of the face and the scalp]. PMID- 12746678 TI - [Congenital hemangiomas]. PMID- 12746679 TI - [Phototests]. PMID- 12746680 TI - [Sclerotherapy of varicose veins of the leg]. PMID- 12746681 TI - [Face-lifting lasers: technical principles and characteristics]. PMID- 12746682 TI - [Lyme's serology]. PMID- 12746683 TI - [Antinuclear antibodies]. PMID- 12746684 TI - [Ursodeoxycholic acid/dapsone interaction]. PMID- 12746685 TI - [The role of the International Committee of Dermatology]. PMID- 12746687 TI - [Neurosurgery for spasticity. Introduction]. PMID- 12746689 TI - [Development and maturation of the pyramidal tract]. AB - The pyramidal tract contains axons that originate from neurons located in layer 5 of the neocortex of the frontal areas 4 and 6 and of the parietal lobe. These neurons are generated during the first half of gestation in humans. The growth of these axons is highly regulated and the mechanisms that control this growth begin to be unravelled. For example, netrins could serve as chemattractants, the adhesion molecule L1 plays a crucial role in the control of axonal decussation at the level of the medulla, the ephrin B3-Eph A4 couple prevents the axons from crossing the midline. During development, the total number of pyramidal axons increases progressively and then decreases by regression of exuberant collaterals. The pyramidal tract is the sole unmyelinated tract in the human spinal cord at birth. This accounts for the protracted central conduction time in newborns. This immaturity of the pyramidal system could explain the existence of specific motor reflexes in newborns (the so-called primary reflexes) that disappear as the pyramidal system matures. PMID- 12746688 TI - [History of neurosurgical treatment of spasticity]. AB - The history of neurosurgery for spasticity is strongly linked to the beginning of neurosurgery. With the discovery of the stretch reflex by Sherrington and the quality of the clinical studies at that time, especially the description of the different kinds of hypertonia by Babinski, the new surgeons of the nervous system started early with interruption procedures on dorsal roots (Foerster) or peripheral nerves (Lorenz, Stoffel). In France, this field of functional neurosurgery grew rapidly. Gros in Montpellier improved the technique of dorsal rhizotomy, while Sindou in Lyons, created the technique of drezotomy after studies on pain mechanisms. The history was then followed in Chicago by Penn and Kroin who developed the technique of intrathecal baclofen which indications are still increasing today. Improvement of knowledge on neurophysiology and control of movement lead to an optimisation of the surgical procedures where French speaking neurosurgery plays an important role. PMID- 12746690 TI - [Anatomical basis of motricity for the study of spasticity]. AB - The spinal motoneurone is under the permanent influence of peripheral afferent fibers, interneurons, and numerous descending projections from supraspinal structures. Motoneuronal activity summarizes these different and convergent modulations at one moment. Spasticity corresponds to exageration of monosynpatic reflex, from IA fiber to motoneuron alpha, associated with spinal hyperexitability. Various lesions of central nervous system give rise to spasticity, specially if they affect supra spinal descending controls, mainly reticulo-spinal tracts. The role of neuronal plasticity to explain the progressive time course of spasticity is also discussed. PMID- 12746691 TI - [Pathophysiology of spasticity]. AB - Lance's definition of spasticity focuses on the exaggeration of the tonic stretch reflex as one component of the upper motor neuron syndrome. In daily practice, many different symptoms are referred to as spasticity. Experimental studies stress the particular role of the premotor cortex and the medial reticular formation for the genesis of spasticity. Physiological studies clearly demonstrate the two components, phasic and tonic, of the stretch reflex. Whatever the pathology, the clinical picture of spasticity seems to depend less upon the etiology of the lesion and more upon its location in the neuraxis. There is a regional organisation of the spinal circuitry according to the function of the segmental nerves and, therefore, a particular clinical presentation related to each spinal segment. The muscular efferents are also heterogeneous and linked to function. This is the fundamental base of focal treatment of spasticity. PMID- 12746693 TI - [Clinical evaluation of spasticity]. AB - Spasticity is one component of the upper motor neurone syndrome. Unlike motor deficit and co-contraction, spasticity is responsive to treatment. Thus it receives more attention than the other features of the upper motor neuron syndrome. In order to treat correctly the patients, we need to know expectations of the patient and family. Detailed clinical evaluation including functional analysis is necessary to determine whether the objectives are reasonable. This evaluation is not always very easy because of complex clinical features. Simply aiming to reduce spasticity is not a final treatment goal. There must be a purpose behind it. It is actually very difficult to evaluate functional effect of spasticity treatments. Few studies show a real functional improvement. Yet patients are satisfied with the results and the goals are often achieved. One explanation is that functional scales usually used do not fit to the spasticity problem. PMID- 12746692 TI - [Pathology of spastic muscles. Study of 26 patients]. AB - Spastic hypertonia is observed in various central neurological diseases. Pathology of spastic muscle is not well known. Tibial selective microsurgical neurotomy is used for the spastic foot treatment. Twenty-six hemiplegic patients, spastic since more than one year, underwent selective tibial microsurgical neurotomy. Biopsies of soleus, lateral and medial gastrocnemius muscles were performed during surgical treatment, after free patient consent and ethical committee approval. The aim of this study was to describe histopathological lesions of spastic muscles, with histological and histoenzymatic techniques. Histological studies revealed myogenic atrophy, concerning both types of fibers or only type II fibers. They also showed type I (slow fibers) to type II (fast fibers) transformation, in the three studied muscles. These results are similar to those described in the literature. PMID- 12746694 TI - [Clinical assessment of spasticity in children]. AB - This article presents the four principle steps of assessment of children with cerebral palsy. Weakness of some muscles groups and spasticity of others and the subsequent functional impairment can be identified by observation. Analytical assessments of spasticity with the Ashworth and Tardieu scales were compared in 30 children with cerebral palsy, mean age 8.5 years. The results pointed out the complimentary nature of these two scales: The Gross Motor Function Measure, which is a validated scale, is specific for children with cerebral palsy. This measure provides a useful assessment of outcome and a convenient means of evaluating the time-course. Joint assessment is also essential to identify muscle contracture. Assessment of spasticity in children with cerebral palsy must be conducted by a multidisciplinary team to take into account all the aspects involved. PMID- 12746695 TI - [Electrophysiological assessment of reflex pathways involved in spasticity]. AB - Various spinal reflexes can be involved in spasticity. Monosynaptic Ia excitation of motoneurons is the most widely studied electrophysiological phenomenon and can be explored by recording the H-reflex, particularly in the soleus muscle. Using conditioning stimuli, the H-reflex technique can be used to study most of the many spinal mechanisms of motor control such as disynaptic, presynaptic, or reciprocal inhibition, post-activation synaptic depression, or facilitation by type I or II muscle afferents. The results obtained in spastic subjects show that the myotactic stretch reflex is exaggerated, but that all the other spinal reflexes are modified. The most recently reported results show an exaggeration of heteronymous facilitation in group II afferents could be one of the main mechanisms leading to spasticity. At the present time, results obtained for the various spinal pathways remain to be compared before concluding about the precise pathophysiology of spasticity, particularly concerning its origin in humans. Despite their complexity, electrophysiological reflex studies offer essential information to reach this goal. PMID- 12746696 TI - [Spastic gait analysis. Contribution of the motion lab]. AB - Etienne-Jules Marey introduced gait motion analysis at the end of the XIXth century. It was rapidly adopted by clinicians and Charcot used it at the beginning of the XXth century in La Salpetriere. Motion analysis was widely used after the first optoelectronic system was built by Furne in 1968. The optoelectronic system calculates the orientation of each body segment in the space after the determination of the space co-ordinates of cutaneous markers placed over them. It is particularly useful for spastic gait. Many disturbances of kinematics and kinetics have been described during spastic gait. They are the consequences of spasticity and other motor and sensory deficits of central nervous system lesions. Motion analysis must be coupled with electromyographic recording of spastic muscles activity which, with kinetic analysis, enables distinguishing the mechanisms underlying gait disturbances. Motion analysis provides clinicians with an indispensable tool for understanding spasticity and evaluating therapeutic efficacy. PMID- 12746697 TI - [Interest of anesthetic blocks for assessment of the spastic patient. A series of 815 motor blocks]. AB - BACKGROUND: The purpose of the study was to emphasize the value of anesthetic blocks in the approach to the spastic patient. The report relates our experience concerning 566 patients (ranging in age from 4 to 72 years, mean 48 years) tested by 815 motor blocks performed within a "spasticity and dystonia evaluation" unit. The spasticity was mainly due to stroke (56%), cerebral palsy (21%) and traumatic brain injury (14%). METHODS: Motor blocks were performed with standardized procedure (specific needle, neurostimulator, localization technique), analytic and functional assessment. RESULTS: The anesthetic was mostly 1% non-adrenalized etidocaine, chosen for its onset and duration of action. Re-injections were few and side effects exceptional. Quality and motor blocks results were technique dependent and required patient cooperation. The spasticity disappeared in blocked muscles. Tardieu and Ashworth modified scale showed constantly decreased spasticity (2 to 3 points) with better sensitivity for the Tardieu modified score. Local anesthetic blocks determined the relative contributions of overactivity and of muscle shortening in the generation of the pathologic posture, the muscle or muscles responsible for the spastic pattern and the level of active performance of the antagonistic muscle. New stability was evaluated by functional assessment of gait posture and prehension. CONCLUSION: At the present time, anesthetic motor blocks represent a necessary and decisive stage procedure as regards spastic patient assessment. This method is particularly useful to anticipate a new functional balance and simulate treatment. Motor blocks provide acute knowledge of the pathological pattern and a better adjustment of therapeutic directions. PMID- 12746698 TI - [Physiotherapy for spasticity]. AB - The aims of physiotherapy techniques used for the treatment of spasticity are to favor sensorimotor recovery and gesture relearning and to lead to an optimal independence in daily life activities. For stroke and head injury patients, there are several techniques sometimes based on opposing principles. The concept of Bobath tries to inhibit the spastic paralysis and the associated reactions to improve the voluntary motricity of limbs with the ultimate goal of enabling exercises in a functional situation, sometimes after a very long period of therapy. On the contrary, according to the concept of Brunnstom, the goal of exercise is to strengthen the spastic paralysis and the associated reactions to enable the upright position and walking as soon as possible. This technique is especially used in very severe deficiencies where the aim is to avoid the bedridden situation. Three active principles can be identified for neurological rehabilitation. Electrical stimulation is not used routinely by rehabilitation teams. It allows to reduce the spasticity of antagonist muscles working against stimulated muscles. It participates in improving the strength of contraction of weak muscles notably in subjects with incomplete paraplegia. Finally, it can be used to improve or replace a functional command (lifting the foot during walking, for example). Nevertheless, electrical stimulation cannot replace basic rehabilitation exercises. PMID- 12746699 TI - [Medical treatment of spasticity]. AB - Spasticity is one of the clinical signs observed after a lesion of the pyramidal tract. Clinical manifestations are polymorphous and depend on the location of the lesion on the pre-motoneuron. Functional consequences are also variable. Only negative effects such as painful spasms, stiffness, distortions, are to be treated. Three different categories of drugs are available: GABA-like (baclofen, benzodiazepine), central alpha 2 agonists (tizanidine, clonidine) and peripheral anti-spastics (dantrolene). Baclofen remains the most commonly used anti-spastic. The preferential indication is spasticity from spinal cord disease, especially when the aetiology is multiple sclerosis. Efficacy of benzodiazepines (diazepam, tetrazepam, clonazepam) is comparable with baclofen; however, side effects (drowsiness) are more frequent. Benzodiazepines are indicated when spasticity is associated with anxiety. Tizanidine is an efficient and well tolerated antispastic. In France, prescription requires a temporary authorization of use. Dantrolen has a peripheral mechanism of action and can be prescribed in the different forms of spasticity. There are other compounds with anti-spastic properties (gabapentine, cyproheptadine, piracetam). Their advantage is rather limited when used alone. Generally, they are administrated in combinaison with usual anti-spastic drugs. PMID- 12746700 TI - [Alcohol neurolytic blocks for pain and muscle spasticity]. AB - Peripheral nerve blockade is one of the therapeutic options for spasticity of various muscles. Percutaneous nerve stimulation allows accurate location of nerves and neurolysis can be performed using intraneural injection of 65% ethanol or 5 to 12% phenol. Spastic contraction of various muscle groups is a common source of pain and disability which prevents efficient rehabilitation. Neurolytic blocks are possible in most of motor nerves of the upper and lower limbs and main indications are spastic sequelae of stroke and spinal trauma but also of multiple sclerosis, cerebral palsy and chronic coma. The use of percutaneous nerve stimulation allows accurate location and four nerves are more frequently treated: pectoral nerve loop, median, obturator and tibial nerves. In patients with spasticity of the adductor thigh muscles, nerve blocks are performed via a combined approach using fluoroscopy and nerve stimulation to identify the obturator nerve. No complications occur and minor side effects are transient painful phenomena during injection. These approaches have proved to be accurate, fast, simple, highly successful and reproducible. Percutaneous neurolytic procedures, should be performed as early as possible, as soon as spasticity becomes painful and disabling in patients with neurological sequelae of stroke, head trauma or any lesion of the motor neurons. PMID- 12746701 TI - [Spasticity and botulinum toxin in 2003. An update]. AB - After the spastic foot in cerebral palsy, there are now wider indications for botulinum toxin injections in spasticity. Post stroke upper limb spasticity has been usefully treated by botulinum toxin in several studies, including double blind placebo-controlled studies. Two serotypes and one serotype B are marketed, with various properties. Botulinum toxin has been studied in multiple etiologies of spasticity. In multiple sclerosis, few studies revealed an efficacy in angulations and comfort. In spinal cord injuries, gait and sphincter disorders can be improved. In post stroke spasticity, lower limb angulations are improved, but gait remained difficult to evaluate. In upper limb spasticity, angulation, function and quality of life were improved in double blind, placebo controlled studies. Comparisons of costs and efficacy are made between botulinum toxin and the other antispastic methods. PMID- 12746702 TI - [Intrathecal baclofen. Experimental and pharmacokinetic studies]. AB - Baclofen, the most effective drug to treat spasticity, is a specific agonist of gamma-aminobutyric acid-B receptors, and is very abondant in the superficial layers of the spinal cord. Given orally, baclofen does not easily penetrate the blood-barrier, and is distributed equally to the brain and spinal cord. After oral administration of baclofen, the drug is resorbed by more than 80-90% in the stomach and bowel and is eliminated by urinary excretion. Failure of oral medication to produce sufficient relief of spasticity is due to the poor passage of the drug across the blood-brain barrier. In animals the concentration in brain is less than 1/10 of the blood levels. The problem of insufficient anti-spastic efficacy (in relation to the rate of side-effects) after systemic medication may be overcome by local application in spinal CSF. Direct intrathecal administration of baclofen in the treatment of severe spasticity was proposed in 1984 by Richard Penn with the objective to carry out a selective spinal distribution of the active principle thus avoiding supraspinal side effects. The pharmacokinetics of baclofen in animal and man after intrathecal administration have been investigated to determine the CSF pharmacokinetic parameters. PMID- 12746703 TI - [Intrathecal baclofen. Literature review of the results and complications]. AB - Intrathecally delivered baclofen has been used as a treatment for severe spasticity since 1984. A systematic literature review was conducted from 1984 to December 2002 to analyze the results of this treatment and to collect data on complications. Studies were included if the following criteria were met: clear selection of patients suffering from spasticity of spinal or cerebral origin, clear measurements of outcome (Ashworth score, Spasm score and/or reflex score and/or functional scales), average follow-up of at least 6 months. Almost all the studies had open follow-up with no control groups (controls were used to examine the effect of test doses of baclofen rather than to assess long-term results). Studies often included heterogeneous patients groups with different causes of spasticity (spinal and/or cerebral etiology) and functional outcome was measured using different scales from one study to another. This literature review shows evidence that intrathecally administered baclofen is effective in reducing the positive signs of spasticity (tone, spasms, reflex activity). Significant reductions in spasm-related pain were noted. The reduction in spasticity led to improvement in ability to transfer and ease of nursing care in the majority of patients. Significant improvements were noted in terms of mobility. Benefits were most notable in bedridden patients who became able to sit in a wheelchair. Many benefited from improved wheelchair mobility, ability to sit down comfortably, and improvement in their ability to transfer. Such benefits were approved by all the patients as an improvement of their quality of life. Ambulatory patients could also benefit from an improved gait but were less often treated because they usually relied upon their spasticity for support during ambulation. Complications were rather rare and mainly were not life-threatening, although there was a high rate of catheter dysfunction (10 to 45%) leading to reoperation. Wound complications were the leading cause of explantation in children with cerebral palsy. Despite the risks, patient satisfaction was high and was related to the improvement of the quality of life. PMID- 12746704 TI - [Intrathecal baclofen and traumatic brain injury. A review]. AB - Severe head injury can induce hypertonia. There are two kinds of hypertonia related to severe injury. The most common one is a spasticity that occurs a long time after the head injury; the second is hypertonia that occurs during the early stage of the post-trauma recovery. Hypertonia is associated with other symptoms and defines post-trauma dysautonomia. Intrathecal baclofen can alleviate spasticity in severe head injury with improvement of the Ashworth score, spasm score or reflex score. But it also improves the number of paroxymal dysautonomic episodes. Real functional improvement is more difficult to show. To be effective, intrathecal baclofen therapy in brain injured patients needs high doses per day, through a spinal catheter in the mediothoracic region, and delivered at least continuously for a few hours to achieve a functional improvement. PMID- 12746705 TI - [Peripheral neurotomies for the treatment of focal spasticity of the limbs]. AB - Introduced by Stoffel in 1912 for the treatment of spastic equinus foot, selective fascicular neurotomy consists in a partial section of the motor collaterals of the muscles presenting excessive spasticity. This peripheral surgery for spasticity is based on two main concepts: 1) Spinal reflexes and muscles are heterogeneous and have a regional organization depending on their role during normal movements. This is the physiological base of focal spasticity and peripheral treatment. 2) There is a differential re-innervation after a partial section, leading to motor recovery without spindles reinnervation (therefore without spasticity). Before performing neurotomies, a careful clinical assessment is required: what kind of spasticity? For what kind of goal (functional or comfort)? The surgery effects can be mimicked by motor nerve blocks (anesthetic drugs) to give to the patient an idea of the expected result. There are many neurotomies, depending on the clinical status, either in the lower or the upper limb. The most frequent is tibial neurotomy for spastic equinus foot. PMID- 12746706 TI - [Percutaneous sacral thermorhizotomy to treat equinism of spastic cerebral palsy children]. AB - We propose to evaluate the effect of sacral percutaneous thermorhizotomy on cerebral palsy children. A prospective study including 29 children followed by a multidisciplinary team was conducted from 1990 to 2000. A thermal radiofrequency lesion of the first sacral root was obtained by percutaneous puncture of the posterior intervertebral foramen with evaluation by stimulation of the best motor response for the minimal threshold (<0.5 volts). The efficacy of the lesion was evaluated by testing and modification of the stimulation threshold obtained (x 2). The results on spasticity was evaluated using the Held score and the functional effect on walking, and classified as "good" (9 cases), "efficacy" (6 cases) and "nil" (4 cases). Indications of the procedure are discussed during the walking acquisition period and growing period of the cerebral palsy child. PMID- 12746708 TI - [Microsurgical drezotomy for spastic limbs]. AB - A microsurgical drezotomy can be useful in some selected cases for the treatment of severe and diffuse spasticity in one or several limbs. This technique has been introduced by M. Sindou in 1972, on the basis of human anatomical studies in the dorsal root entry zone (DREZ). The aim of surgery is to interrupt preferentially the peripheral inputs sustaining myotatic and polysynaptic reflexes which are exaggerated in spasticity, and also nociceptive fibers in the lateral part of the DREZ, whilst sparing most of the medial tactile and kinestetic fibers for the dorsal funiculus. The authors report a retrospective review of 269 patients with severe spasticity in one upper limb (94 cases) treated by a cervical drezotomy, and in one (17 cases) or two lower limbs (158 cases) treated by a lumbosacral drezotomy. The place of this ablative technique inside the neurosurgical armament against spasticity is discussed. PMID- 12746707 TI - [Dorsal rhizotomies in children]. AB - Described for the first time by Foerster in 1908, dorsal rhizotomies are widely performed in spastic children, mainly in diplegics. The different techniques are presented: selective then sectorial rhizotomies developed by Gros in Montpellier, functional rhizotomies developed by Fasano. Literature analysis show a decrease in spasticity for 75% of the cases with a functional improvement in the less affected patients, i.e. those with still effective motor function. The most recent studies are devoted to the improvement of the spinal approach by laminotomies and show an increased interest for preoperative monitoring to limit the extent of the sections. In our experience, this surgery should be limited to children with uncontrolled spasticity which interferes with locomotor performances. PMID- 12746710 TI - [Orthopedic treatment of consequences of upper limb spasticity]. AB - Orthopedic surgery for upper extremity spasticity is based on a precise clinical evaluation with motor and sensitive and cognitive factors. Use of electromyography and lidocain or toxin is necessary. Surgical aims are classified as hygienic or functional. Surgical procedures are divided into neurectomies, muscle, tendon and joint procedures. Results, in spite of heterogeneous populations and nonstandardized assessments, are described about shoulder, elbow and hand. PMID- 12746711 TI - [Physiology and evaluation of overactive bladder]. AB - Detrusor overactivity is a urodynamic observation characterized by involuntary detrusor contractions during the filling phase which may be spontaneous or provoked. There are certain patterns of detrusor overactivity. Phasic detrusor overactivity is defined by a characteristic wave form and may or may not lead to urinary incontinence. Terminal detrusor overactivity is defined as a single involuntary detrusor contraction occurring at cystometric capacity, which cannot be suppressed. Neurogenic detrusor overactivity is qualified as neurogenic when it is a relevant neurological condition (old term is "detrusor hyperreflexia"), idiopathic detrusor overactivity when there is no defined cause (this term replaces "detrusor instability"). In neurogenic patients, detrusor overactivity is secondary to various pathophysiologic factors: interruption of inhibitor pathways issued from cerebral regions, activation de novo of vesical C reflex mediated by unmyelinated capsaicin fibers, and ultrastructural modifications of bladder urothelium. Bladder overactivity treatment is necessary to avoid renal complications and improve quality of life of neurogenic patients. PMID- 12746709 TI - [Surgical treatment of orthopedic deformities due to spasticity in the lower limb]. AB - Orthopedic deformities in the lower limb concern all joints (hip, knee, ankle, foot) with a wide range of clinical forms. Spasticity, contracture, stiffness, laxity, neurological deficit are assessed to establish the surgical procedure. Surgical techniques are adapted to the goals that are detailed with the patient and his family: standing, transferring, walking, hygiene, devices (shoes, orthosis, canes, wheelchair). Surgical procedures can associate: lengthening of contractured muscles (tenotomy with or without sutures, fractional lengthening at the musculo-tendinous junction or desinsertion), strengthening of antagonists (passive or active tendon transfer) and correction of joint deformity (arthrolysis, arthrodesis, arthroplasty). In adults, the most common deformities are the equinus or equinovarus foot, toe curling, hip adductum, knee flessum. Talus or knee recurvatum are less frequently observed. The association of various deformities raises questions concerning the hierarchy of surgical procedure, from an anatomical point of view (do we start with proximal or distal joint first?) as from chronological concerns (shall we do one or more procedures?). Pluridisciplinary assessment using neurological anesthetic blocs and dynamic EMG or gait analysis is necessary to detail the aims of surgery and choose the surgical procedures. PMID- 12746712 TI - [Effect of spasticity on genitosexual and bladder-sphincter function]. PMID- 12746713 TI - [The medical treatment of overactive bladder]. AB - Overactive bladder, very frequent in neurological disorders, leads to very distressing symptoms such as urgency, frequency and incontinence which may dramatically impair the patient's quality of life. The medical approach is essentially pharmacological but the management of the nociceptive factors must not be neglected. In the mild urinary dysfunctions, bladder training can be advised. The pharmacological treatment aims at reducing the parasympathetic activity or at deafferenting the bladder. The antimuscarinic agents are an essential part of the treatment. Oxybutynin is the most widely used medication but recent agents like tolterodin have a better tolerability. Other drugs can also be used such as desmopressin, flavoxate. New molecules are under experiment (darifenacin). In case of troublesome side-effects or resistance to oral medications, local treatments are considered. Intravesical oxybutynin has been tried but has a short-lived efficacy. Intravesical instillation of capsaicine or resiniferatoxin blocks C-fibres afferents and leads to clinical and urodynamic improvement. Recently, injections of botulinum-A toxin in the detrusor have been advocated aiming at blocking the transmission of parasympathetic impulse. The first studies report encouraging results. All these local treatments resulting in bladder paresis are recommended for patients performing self-catheterization. Should these treatments fail, other therapeutic approaches are considered such as intrathecal treatment, neuromodulation, before deciding on neurosurgical or urosurgical procedures. PMID- 12746714 TI - [Sacral root neuromodulation for the treatment of urinary incontinence reported to detrusor hyperactivity]. AB - BACKGROUND AND PURPOSE: This paper reviews therapeutic sacral neuromodulation for treating urinary urge incontinence related to detrusor hyperactivity. METHODS: We reported data from our department and from the international literature on topics such as the physiological basis of neuromodulation, techniques of testing, chronic implantation and clinical results. RESULTS: In an intention to treat analysis , neuromodulation results varied from 21.5 to 25% globally. On implanted patients, the response rate varied from 40 to 88% and was stable. Sub-chronic test morbidity was very rare. Surgical revision rate was reported from 6.25 to 37.7%. CONCLUSIONS: Neuromodulation strongly ameliorates approximately a third of all the patients with urge urinary incontinence due to detrusor hyperactivity. New technical improvements should lead to better results in the future. PMID- 12746715 TI - [Anterior sacral root stimulation with dorsal rhizotomy (Brindley technique)]. AB - Patients with spinal cord lesions are exposed to vesico-sphincter dysfunctions which can aggravate the vital or functional prognosis. The pathophysiological characteristics of these neurogenic bladders explains the usefulness of surgical treatment. The objective of the Brindley technique is to improve both voiding and effective continence. Any patient with a stable supra-sacral spinal cord lesion (paraplegia, tetraplegia) with a reflex bladder (incontinence, vesico sphincter dyssynergia resistant to medical treatment with the risk of upper urinary tract involvement) can benefit from the Brindley technique. The electrodes are placed on the anterior sacral roots in order to obtain the desired micturation. Posterior sacral rhizotomies are indispensable to the technique as they suppress detrusor and sphincter hyperreflexia and improve continence thereby protecting the upper urinary tract. About 90% of the patients considered have an improved quality of life after implantation of the Brindley stimulator. The bladder capacity was constantly improved and the majority of the patients become continent. Micturation was excellent with low residual volume and low rate of urinary tract infections. Complications are analyzed in this review of the literature. The Brindley technique is an excellent alternative to medical treatment in these highly distressed patients. It restores satisfactory continence and improves psychological as well as economical constraints related to auto/hetero catheterisations performed several times a day. PMID- 12746716 TI - [Intrathecal therapy for bladder hyperreflexia]. AB - Intrathecal clonidine was tested for the control of bladder hyperreflexia resistant to pararsympathicolytic drugs in spinal cord injured patients. Urodynamic parameters were significantly improved after acute bolus of intrathecal low doses of clonidine. Unfortunately, the chronic intrathecal infusion of clonidine induced cardiovascular side-effects. Intrathecal baclofen did not modified significantly neurogenic bladder dysfunction from spinal lesion. But intrathecal baclofen modified penile erection quality and impaired the ejaculation reflex induced by penile vibratory stimulation in the same population. PMID- 12746717 TI - [Microsurgical sacral drezotomy for the treatment of hyperactive bladder]. AB - In paraplegics who can benefit from bilateral microsurgical L2-S1 drezotomy for the treatment of harmful spasticity involving the two lower limbs, surgery can be extended to the segments S2-S4 for the treatment of an associated hyperactive bladder. The surgery in the dorsal root entry zone (DREZ) is designed to preferentially interrupt nociceptive and myotatic inputs from the detrusor muscle reaching the sacral spinal segments S2, S3, S4. This work reports a prospective study of 38 patients who underwent bilateral S2, S3, S4 drezotomy with a follow up of 18 months on average. All these patients were unable to have voluntary miction and had sexual dysfunctions. Urge incontinence was present in all of them. A hyperactive bladder was present at the preoperative urodynamic study in all patients. A vesico-ureteral reflux was demonstrated in 13 cases (34%) during cystography. A postoperative urodynamic study was performed at 6 months on average and demonstrated disappearance of hyperactivity of the detrusor muscle in 82% of the cases and increased bladder capacity in 63%, corresponding to patients without a preoperative fibrotic detrusor. They were no cases of postoperative vesico-ureteral reflux at cystography. At 18 months, the urine leakages have disappeared in 89% and decreased to less than half per day in the other 11%. As expected, all the patients required postoperative permanent or intermittent catheterization. The morbidity was acceptable in these severely impaired patients. To conclude, a bilateral sacral (S2-S4) microsurgical drezotomy can be performed in addition to a lumbosacral drezotomy to treat an active bladder associated with harmful spasticity in both legs. The sacral microsurgical drezotomy allows to achieve bladder continence and to protect the upper urinary tract PMID- 12746718 TI - [Indications of neurosurgical procedures for spasticity]. AB - Neurosurgery for spasticity requires perfect knowledge of all the mechanisms involved in the control of movement. Spasticity should be clearly checked in all these aspects: phasic, tonic and flexor reflexes. Its contribution to the patient's handicap should be assessed. Motor block or intrathecal injections of baclofen are useful tests to evaluate the surgical indications for well defined goal and understood by the patient himself. The choice of the procedure depends on the patient and the type of spasticity. Post-operative assessment is very important to improve the surgical procedures in the future, going towards motor restoration. PMID- 12746719 TI - [Neurosurgical treatment of spasticity: indications in children]. AB - Today, we have several efficient neurosurgical treatments of spasticity in children with cerebral palsy. A good indication is possible only if a consensus about the goal of the surgery is found between the surgeon, the child and his/her family, and the reeducation team. This goal is not always functional. Clinical examination is not limited to the analytical assessment of spasticity, but must take into account the general and orthopedic state of the child, and his/her functional evolution, cognitive abilities, habits and general environment. The struggle against spasticity is part of a therapeutical programme which extends over several years. It must be considered before muscular contractures. On lower limbs, in the cases of general spasticity, we propose posterior rhizotomy or intrathecal baclofen administration. Posterior rhizotomy is proposed when a more aggressive intervention is preferred for some muscular groups or when the child's general environment does not allow for the medical care imposed by intrathecal administration. In the case of localized spasticity, botulinum toxin injection permits delaying until the child reaches the age for selective neurotomy. On upper limbs, in children with quadriplegia the indication is essentially physical and occupational therapy. We cannot count on the positive side effects of rhizotomy or intrathecal administration of baclofen on the upper limbs. In children with hemiplegia, with localized or global spasticity of the upper limb, botulinum toxin is proposed as the first form of treatment. Neurotomy or rhizotomy can follow toxin, according to the efficacy of the toxin. PMID- 12746720 TI - [Spasticity: today and tomorrow]. AB - History of the spasticity is evoked including its physiological and medical aspects. The reduction of functional capacity of the patient experiencing spasticity is, for the author, one of the major duties of the medical staff involved in the treatment of this symptom. Some aspects of the measure of the reduction of functional capacity due to this symptom are described. Its seems to the author that the new treatments (neuronal graft.) should increase the number of patients to be treated for spasticity. PMID- 12746721 TI - [Accuracy of intraoperative diagnosis in central nervous system tumors: report of 1315 cases]. AB - Clinicians order neurosurgery frozen sections in order to answer three questions: is the specimen tumor tissue? is it benign or malignant? what is its histological type? We studied the diagnostic accuracy of 1 315 frozen sections of central nervous system tumors, performed between 1988 and 1999, and compared it with data in the literature. Agreement between intraoperative and paraffin-section diagnosis was 96.6% (rate of error: 3.4% for the question tumor tissue or not). The answer was concordant in 92.6% with a 7.4% rate of error for tumor malignancy or benignity. Exact histological concordance was 87.6%. The most frequent errors in histological typing concerned gliomas, hemangioblastomas and metastasis. Our results emphazise the reliability of intraoperative frozen sections in Neurosurgery and the importance of close collaboration between clinicians, radiologists and pathologists. PMID- 12746722 TI - [A retrospective study of 32 lumbar synovial cysts. Clinical aspect and surgical management]. AB - We report a retrospective analysis of a series of 32 cases of lumbar synovial cysts. Clinically, two populations were identified: patients who presented radicular pain mimicking the clinical diagnosis of disk herniation (group I) and patients who presented neurogenic claudication consistent with a diagnosis of lumbar spinal stenosis (group II). The radiological work-up consisted in standard X-rays and CT-scan for all patients. MRI (magnetic resonance imaging) was performed in 14 patients. All the patients were operated on. The surgical technique consisted in foraminotomy for patients in group I and more or less extensive laminectomy for patients in group II. Functional outcome was marked by a significant improvement in 96.9% of the patients and those who presented a motor deficit recovered in 83% of the cases. Based on the findings in this series, we discuss the clinical aspects and the therapeutic management of this pathological entity. PMID- 12746724 TI - [Thoracic spinal stenosis. Clinical and surgical management. Study of 5 cases observed in the Abidjan department of Neurosurgery]. AB - BACKGROUND: Spinal cord compression resulting from thoracic spinal stenosis is relatively frequent in Japan but rare in Europe and little know in Africa. OBJECTIVE: We report five cases of thoracic myelopathy caused by thoracic stenosis diagnosed in black African people. METHODS: This was a retrospective observational study of patients treated in the Abidjan department of Neurosurgery from 01.01.1996 to 31.12.1998. The spinal cord compression resulting from thoracic spinal stenosis was confirmed by myelography or myeloscan. The Japanese orthopaedic association (JOA) score was used to evaluate patient outcome. RESULTS: All of these patients were black Africans. The four men and one woman were aged 55, 47, 53, 52, and 60 years. The clinical presentation of thoracic spinal cord compression was confirmed by myelography in five patients and by myeloscan in four. These investigations were useful for guiding surgical treatment. Outcome was favorable in all cases. CONCLUSION: Signs of thoracic spinal cord compression in adults should suggest the diagnosis of thoracic spinal stenosis. PMID- 12746723 TI - [Use of polyester urethane (Neuro-Patch) as a dural substitute. Prospective study of 70 cases]. AB - Between October 1995 and March 1998, 70 patients were treated with a microporous polyester urethane dura substitute (Neuro-Patch), after brain or spinal surgery. These patients were assessed clinically and radiologicaly 10 days, 6 weeks and 1 year after surgery. Radiological evaluation used CT scan or MRI. All dura substitutes were fixed by continuous suture to the surrounding dura-mater. We studied the handling properties, the incidence of infection and of CSF leakage. Eleven patients underwent craniotomy again. This gave us the opportunity to examine the adhesion to the brain tissue and the integration of the dura substitute. Six sheets underwent histological examination. Our results show good handling properties of the material; 3 infections; 6 out of 9 radiological CSF leakage occurred from infratentorial surgery. During reoperation, no adhesion to the brain tissue or injury to the brain while detaching the dura substitute was noticed. An excellent histological integration was observed: pores of the Neuro Patch were colonized by fibroblasts synthesizing collagen, and there was no immune or inflammatory reaction, with an actual 4 to 6 years follow-up. A Neuro Patch can therefore be recommended as a dura substitute to repair spinal or cranial dural defects. PMID- 12746725 TI - [Glutamate in traumatic brain injuries]. AB - During the past years, studies dealing with the pathophysiology of brain injuries have focused on certain agents that seem to be particularly involved in secondary brain lesions. Of them, some amino acids, and particularly glutamate, appears to deserve special attention. We discuss the role of glutamate neurotransmitters, review glutamate metabolism and actions, and then review in the pertinent literature devoted to the role of glutamate in the onset of brain injuries secondary to head trauma. PMID- 12746726 TI - [Post-traumatic exophthalmos caused by a subperiosteal hematoma of the orbit]. AB - We report a case of a 27-year-old man who developed major exophthalmos associated with ophthalmoplegia occurring 48 hours after head trauma. CT scan showed an orbital roof fracture and a subperiosteal hematoma. Surgical evacuation of the hematoma and reconstruction of the orbital roof was performed via a frontal craniotomy. Two months later, the ophthalmologic examination was normal. We stress the importance of early diagnosis and prompt surgical treatment in such an unusual complication of head trauma. PMID- 12746727 TI - [Spontaneous otogenic pneumocephalus]. AB - Otogenic pneumocephalus is uncommon. Typical causes include trauma, tumor, infection, and nasosinusal or mastoid surgery but spontaneous otogenic pneumocephalus is very exceptional. We report a case of spontaneous otogenic pneumocephalus located in the left temporal lobe revealed by sudden onset aphasia. The temporal pneumatocele was cured through an epidural subtemporal approach with needle puncture of the aeroma and duroplasty. Two years after surgery, no recurrence was observed and the patient remained symptom free. PMID- 12746728 TI - [Spontaneous intrasphenoidal encephalocele. Report of two cases revealed by rhinorrhea]. AB - We report two cases of spontaneous anteromedial temporo-sphenoidal meningo encephalocele. The first patient was a 67-year-old man with a history of treated epiglotic cancer 6 years ago, was referred to us for CSF rhinorrhea. Lateral sphenoidal CSF fistula was confirmed by CT cisternography. Four lumbar taps were performed and rhinorrhea stopped for 2 years. At relapse of rhinorrhea, the patient was operated on via a subtemporal approach. The dural defect was filled by an intradural patch of pericranium. Four years later patient showed no evidence of cerebrospinal fluid leakage. The second patient was a 54-year-old woman referred to us after a one year history of CSF rhinorrhea. CT cisternography confirmed lateral sphenoidal CSF fistula. The patient was operated on via a subtemporal approach. The dural defect was filled by a patch of pericranium. Three years later the patient had no evidence of cerebrospinal fluid leakage. Twenty-one cases of anteromedial temporosphenoidal meningo encephaloceles have been reported in the literature. They probably are of a developmental origin. Some cases have been successfully treated via a transsphenoidal route. However surgical failures related to this approach can be due to the lack of control of the lateral recess of the sphenoid sinus. Therefore, the frontotemporal approach seems more adequate but the transphenoidal endoscopic approach will probably be used in the future. PMID- 12746729 TI - [Intracerebral granulocytic sarcoma. A case report]. AB - Granulocytic sarcoma is a tumor composed of proliferating myeloblastic cells, generally found in the orbit. A brain localization is rare. We report the case of a 11-year-old boy treated in our unit for acute myeloblastic leukemia (AML 4 Eo. FAB). After 21 months of complete remission, he developed headache and facial palsy. The CT scan visualized the presence of two frontal and occipital masses. The spinal tap revealed blastic cells in the CSF. The study of the bone morrow showed medullar relapse. A new medullar and cerebro-meningeal remission was obtained with chemotherapy and radiotherapy. CSF and the bone marrow studies can help avoid stereotaxic biopsy can be avoided in this type of tumor PMID- 12746730 TI - Irritable bowel syndrome genophenomics: correlation of serotonin-transporter polymorphisms and alosetron response. PMID- 12746731 TI - Optimization and individualization of thiopurine therapy in inflammatory bowel disease: the great variability among drug responses. PMID- 12746732 TI - Pharmacoproteomics in drug development. AB - The field of proteomics is taking on increased significance as the relevance of investigating and understanding protein expression in disease and drug development is appreciated. Recent advances in proteomics have been driven by the availability of numerous annotated whole-genome sequences and a broad range of technological and bioinformatic developments that underscore the complexity of the proteome. This review briefly addresses some of the various technologies that comprise Expression Proteomics and Functional Proteomics, citing examples where these emerging approaches have been applied to pharmacology, toxicology, and the development of drugs. PMID- 12746733 TI - Single nucleotide polymorphism genotyping: biochemistry, protocol, cost and throughput. AB - The large number of single nucleotide polymorphism (SNP) markers available in the public databases makes studies of association and fine mapping of disease loci very practical. To provide information for researchers who do not follow SNP genotyping technologies but need to use them for their research, we review here recent developments in the fields. We start with a general description of SNP typing protocols and follow this with a summary of current methods for each step of the protocol and point out the unique features and weaknesses of these techniques as well as comparing the cost and throughput structures of the technologies. Finally, we describe some popular techniques and the applications that are suitable for these techniques. PMID- 12746734 TI - Cebus apella, a nonhuman primate highly susceptible to neuroleptic side effects, carries the GLY9 dopamine receptor D3 associated with tardive dyskinesia in humans. AB - Tardive dyskinesia (TD) is a severe side effect of traditional neuroleptics affecting a considerable number of schizophrenic patients. Accumulating evidence suggests the existence of a genetic disposition to TD and other extra pyramidal symptoms (EPS) most strongly linked to a ser/gly polymorphism in position 9 of the D3 dopamine receptor gene (DRD3). The Cebus apella monkey is the favored animal model to study TD and other EPS because of its high susceptibility to side effects of neuroleptics. We therefore determined the sequence of the DRD3 gene in this species and compared it with that of humans. We found that the highly TD susceptible C. apella monkey (n=21) carries the gly9/gly9 DRD3 genotype that has been associated with TD in humans. Contrarily, C. apella did not carry the ser23 5HT2C allele that has been reported to increase TD susceptibility in humans. PMID- 12746735 TI - Antidepressant-induced mania, rapid cycling and the serotonin transporter gene polymorphism. AB - The genes involved in the serotonin system are major candidates in association studies on affective disorders and responses to antidepressants. We studied a functional polymorphism of the serotonin transporter (5-HTT) gene (a 44 bp insertion/deletion in the 5-HTT-linked polymorphic region (5-HTTLPR)) and lifetime history of antidepressant-induced mania (AIM) in a population of 305 patients with bipolar affective disorder. AIM was defined using a broad definition and a restrictive definition. No association was found between the "s" allele of the 5-HTTLPR and AIM for either definition. However, we found an association between the 5-HTTLPR and lifetime history of rapid cycling in a subsample of patients (for allele and genotype distributions: exact probability, p=0.0009 and chi(2)=9.4; df=1; p=0.002, respectively). These results may help to explain the conflicting association results obtained with the 5-HTT gene polymorphism, in particular with AIM. Indeed, the precise phenotype associated with the 5-HTT gene is unclear. The association between the "s" allele and rapid cycling may provide further evidence for an association between the 5-HTTLPR "s" allele and a pattern of affective instability. PMID- 12746736 TI - Pharmacokinetics and QT interval pharmacodynamics of oral haloperidol in poor and extensive metabolizers of CYP2D6. AB - We studied the pharmacokinetics and QT interval pharmacodynamics of a single 10 mg dose of oral haloperidol in a randomized, double-blind, placebo-controlled, crossover trial of healthy poor (PMs) and extensive (EMs) metabolizers of CYP2D6. There was a statistically significant greater mean QT(c) on haloperidol (421.6+/ 20.1 ms) than on placebo (408.4+/-18.5 ms, P=0.0053) occurring 10 h post haloperidol/placebo administration. Men and women had similar ranges of QT(c) changes from placebo. Despite a statistically significant greater mean elimination half-life (19.1+/-3.6 vs 12.9+/-4.0 h, P=0.04) and lower mean apparent oral clearance (12.8+/-4.1 vs 27.0+/-11.3 ml/min/kg, P=0.02) of haloperidol in CYP2D6 PMs than in EMs, this exposure change did not translate into marked QT(c) changes from baseline that could be considered clinically important. Although the magnitude of the mean QT(c) prolongation on haloperidol relative to placebo is relatively small, it may assume significance in the presence of other risk factors for QT prolongation. PMID- 12746737 TI - Soy and social stress affect serotonin neurotransmission in primates. AB - Stress and sex steroidal milieu can each influence mood in women. The purpose of this study was to compare the effect of long-term conjugated equine estrogen (CEE), soy phytoestrogen (SPE), and social subordination stress on dorsal raphe serotonin neurotransmission of ovariectomized cynomolgus monkeys. Tryptophan hydroxylase (TPH) and serotonin reuptake transporter (SERT) protein content were determined, and the in vitro degradation of macaque SERT protein was examined in the presence and absence of protease inhibitors, serotonin (5-HT), and citalopram. Like CEE, SPE increased TPH protein levels. Social subordinates had markedly lower TPH protein levels than dominants regardless of hormone replacement. Therefore, these two variables had independent and additive effects. CEE and SPE increased SERT, and social status had no effect. Thus, the hormone induced increase in SERT was accompanied by increased 5-HT synthesis and neuronal firing, which appears biologically reasonable as 5-HT prevented SERT degradation in vitro. PMID- 12746738 TI - Laminectomy for cervical myelopathy. AB - STUDY DESIGN: Cervical laminectomy with or without fusion, or laminoplasty, successfully address congenital or acquired stenosis, multilevel spondylosis, ossification of the posterior longitudinal ligament (OPLL), and ossification of the yellow ligament (OYL). To optimize surgical results, however, these procedures should be applied to carefully selected patients. OBJECTIVES: To determine the clinical, neurodiagnostic, appropriate posterior cervical approaches to be employed in patients presenting with MR- and CT-documented multilevel cervical disease. To limit perioperative morbidity, dorsal decompressions with or without fusions should be performed utilizing awake intubation and positioning and continuous intraoperative somatosensory-evoked potential monitoring. SETTING: United States of America. METHODS: The clinical, neurodiagnostic, and varied dorsal decompressive techniques employed to address pathology are reviewed. Techniques, including laminectomy, laminoforaminotomy, and laminoplasty are described. Where preoperative dynamic X-rays document instability, simultaneous fusions employing wiring or lateral mass plate/screw or rod/screw techniques may be employed. Nevertheless, careful patient selection remains one of the most critical factors to operative success as older individuals with prohibitive comorbidities or fixed long-term neurological deficits should not undergo these procedures. RESULTS: Short- and long-term outcomes following dorsal decompressions with or without fusions vary. Those with myelopathy over 65 years of age often do well in the short-term, but demonstrate greater long-term deterioration. Factors that correlated with greater susceptibility to deterioration include advanced age (>70 years at the time of the first surgery), severe original myelopathy, and recent trauma. CONCLUSIONS: Success rates of laminectomy with or without fusion, or laminoplasty may be successfully employed to address multilevel cervical pathology in a carefully selected population of patients. PMID- 12746739 TI - Pediatric spinal cord injury in Sweden: incidence, etiology and outcome. AB - STUDY DESIGN: Retrospective descriptive study. OBJECTIVES: To assess incidence, causes and early outcome of traumatic spinal cord injury (SCI) among children in Sweden, thereby identifying high-risk groups and situations as a basis for preventative measures and improved care. METHODS: Data from population registers, County Habilitation Centers as well as from informal sources were used to estimate the incidence of SCI in Sweden during the years 1985-1996 among children aged 0-15 years. Contacts with the treating hospitals, reviews of medical records and/or personal interviews were used to verify primary data. In total, 92 children were thus identified. RESULTS: The incidence was found to be 4.6/million children/year (95% CI 3.6-5.5). When excluding prehospital fatalities, the incidence was 2.4 (95% CI 1.8-3.1). The main cause of injury among fatalities was traffic accidents. Associated injuries occurred in 41% of the children. Among survivors (10-15 years), sports-related injuries (43%) were as common a cause as traffic accidents (39%). The survivors were treated in 18 different hospitals. CONCLUSION: Pediatric SCI in Sweden is rare, presumably because of effective primary prevention. Preventative measures should be further differentiated for each age group of children in accordance with their differing risk profiles. In contrast to the effective preventative measures that have been implemented in Sweden, care of these patients is still too fractionated and decentralized for sufficient specialization to emerge. PMID- 12746741 TI - Dysphagia in patients with acute cervical spinal cord injury. AB - STUDY DESIGN: Longitudinal observational. OBJECTIVES: (a) To establish a reliable and feasible method to indicate the presence and severity of dysphagia and (b) to establish a course of treatment in individuals presenting with cervical spinal cord injury (CSCI). SETTING: Spinal Cord Injury Center, Werner Wicker Klinik, Bad Wildungen, Germany. PATIENTS AND METHODS: This is a cross-sectional study of 51 patients consecutively admitted to the Intensive Care Unit of the SCI in-patient service. They were subjected to neurological and fiberoptic endoscopic examination of swallowing (FEES). Data concerning artificial respiration, presence of tracheostomy, oral or non-oral feeding were obtained from the medical charts. Statistics were carried out by a calculation of a nonparametric correlation (Spearman). RESULTS: Five levels of dysphagia could be distinguished. At levels 1 and 2, patients presented with a severe impairment of swallowing, in level 3 aspiration was met by a powerful coughing reflex, level 4 comprised a laryngeal edema and/or a mild aspiration of fluids only and at level 5 laryngeal function was not compromised. On admission, 20 patients with CSCI presented with mild (level 4), eight with moderate (level 3) and 13 with severe dysphagia (levels 1 and 2). In 10 no signs of dysphagia could be detected. After treatment, level 1 was no longer detected, one patient showed level 2, two patients showed level 3, all other patients showed only mild or no signs of dysphagia any longer. CONCLUSIONS: Dysphagia of various severities was present in the majority of these patients with CSCI together with respiratory insufficiency. FEES allows for the detection and classification of dysphagia as well as for an evaluation of the therapeutic management. Under interdisciplinary treatment the prognosis of dysphagia is good. PMID- 12746740 TI - Pediatric spinal cord injury in Sweden; how to identify a cohort of rare events. AB - STUDY DESIGN: Register study enhanced and verified by medical records and personal interviews and examinations. SETTINGS: Sweden. OBJECTIVES: To define a method of identifying a study population of rare events. To point out the relative importance of every step, an example is given of identifying persons who sustained traumatic spinal cord injury (SCI) in childhood. METHODS: Cases were identified in seven steps that all needed to be fulfilled, from definition of selection criteria through combination of several data sources, to the use of several verification methods. RESULTS: Initial screening by registers identified 384 possible cases, which however were found by subsequent analysis to include a large number of incorrect cases. At completion of all analytic steps, 35 living cases could be fully verified and 14 deceased cases could be partially verified. CONCLUSIONS: Registers offer a practical initial source for study population identification. The screening of International Classification of Diseases codes defining SCI only included less than 30% of 'true' SCIs. Subsequently, further refinement and quality control is necessary in order to ensure validity. Such further verification is time-consuming, but nevertheless necessary in order to verify a true cohort. PMID- 12746742 TI - Predicting normal lung function in patients with childhood spinal cord injury. AB - STUDY DESIGN: A prospective observational study. OBJECTIVES: To compare the height and arm span measurements in childhood spinal cord injured (SCI) people and examine the subsequent effect on calculating the predicted lung function using standard formulae and to discuss which of the two measurements is the most appropriate to use in these formulae. SETTING: National Spinal Injuries Centre, Stoke Mandeville Hospital, Aylesbury, UK. METHOD: A total of 12 children had lung function tests performed and at the same time had height and armspan measured. The predicted lung function was calculated twice; once using height and then using arm span and compared. The actual lung function test results were expressed as percentage of the two predicted values, respectively, and compared. RESULTS: The difference between the mean height (1499 mm) and arm span (1649 mm) measurements was significant (P<0.001). In all cases, the arm span measurement was greater than the height. The two predicted lung function values (one calculated using height and the other armspan) were significantly different (P<0.001). When lung function test results were expressed as percentage of the two predicted values they gave a very different interpretation of the results. The actual performance was much lower than the predicted values if arm span, rather than height, was used in prediction equations. CONCLUSION: In childhood SCI, the difference in height and arm span is significant. This affects the predicted lung function values significantly and thus changes the interpretation of the lung function test results. The most appropriate measurement to use in prediction equations (height or arm span) in these subjects is yet to be decided. PMID- 12746743 TI - Chondroblastoma of the lumbar spine with cauda equina syndrome. AB - STUDY DESIGN: Case report. OBJECTIVE: To describe the clinical presentations, radiological features and clinical progress of a rare case of chondroblastoma of the lumbar spine. SETTING: Regional Hospital, Hong Kong, China. METHOD: A 54-year old male patient presented with low back pain and left sciatica. X-ray and MRI revealed tumour infiltration of the fifth lumbar vertebrae and left paraspinal muscles, which was found to be a chondroblastoma by repeated open biopsies. The tumour was removed surgically by combined anterior and posterior approaches, followed by spinal fusion and instrumentation. RESULTS: The anterior tricortical bone graft was complicated with fracture and nonunion. Surgical re-exploration confirmed local recurrence of tumour macroscopically and histologically. The patient developed symptoms and signs of cauda equina syndrome gradually despite repeated surgery and irradiation. The patient eventually died of complications of local recurrence and neurological deficit at 3 years and 8 months after the first operation. CONCLUSION: This is the first case report of chondroblastoma of the lumbar spine. The clinical profile of this patient and the evidence from the literature review suggests that spinal chondroblastoma has a very aggressive behaviour with high recurrence and mortality rate. PMID- 12746744 TI - Gluteal neuromuscular stimulation in therapy and prophylaxis of recurrent sacral pressure ulcers. AB - STUDY DESIGN: Case study. OBJECTIVE: Positive influence of electrostimulation on the healing process of the gluteal decubital ulcers. SETTING: Department of Neurosurgery, University of Cologne, Germany. METHOD: The present study reports on the effects of the electrostimulation of the floor-of-the-pelvis-muscles by means of an anal electrode. This procedure was shown to have a positive influence on the healing process of the gluteal decubital ulcers sustained by a patient with incomplete sensomotor paraplegia. Apart from the contraction of the floor-of the-pelvis-muscles, we observed a contraction of the gluteal muscles on both sides. RESULTS: The ulcers, which had been resisting conventional treatments for months, showed signs of a beginning healing tendency. After 4 weeks both ulcers were completely healed up. CONCLUSION: Because of its easy handling and its good amicability, electrostimulation of the gluteal region - one of the most common localisations of pressure-caused ulcers - by means of an anal electrode might be put to good effect even in prophylaxis in the treatment of paraplegic patients. PMID- 12746745 TI - Penile urethral obstruction in a subject with spinal cord injury. AB - STUDY DESIGN: Single case report. OBJECTIVE: To report a case of urethral obstruction because of calculus in a subject with spinal cord injury (SCI). SETTING: Bangalore, India. CASE REPORT: A 25-year-old man sustained complete SCI at C(6) level following a road traffic accident. After 14 months, while on self intermittent catheterization he noticed difficulty in introducing catheter and acute retention of urine. X-ray examination revealed a calculus in the penile urethra at the level of the glans penis. CONCLUSION: Impaction of calculi in penile urethra, although rare, can manifest with acute painless urinary retention in patients with SCI. PMID- 12746746 TI - Atrial fibrillation: emerging possibilities for drug treatment: an overview of current opportunities and recent developments. PMID- 12746747 TI - Atrial electrophysiology and mechanisms of atrial fibrillation. AB - Atrial fibrillation is the most common cardiac arrhythmia in clinical practice, and its management remains challenging. A solid understanding of the scientific basis for atrial fibrillation therapy requires insight into the mechanisms underlying the arrhythmia, about which an enormous amount has been learned over the past 10 years. The basic information presently available about atrial fibrillation mechanisms is reviewed. The particular properties of normal atrial electrophysiology are discussed, including salient ionic determinants of the atrial action potential and key anatomic features. Reviewed are three crucial arrhythmia mechanisms long held to be involved in atrial fibrillation: 1) rapid ectopic activity, 2) single-circuit reentry with fibrillatory conduction, and 3) multiple-circuit reentry. The determinants of each and the evidence for their involvement in clinical and/or experimental atrial fibrillation are noted. The physiological consequences, various contributing mechanisms, and clinical implications of the role of atrial-tachycardia remodeling are analyzed. Atrial tachycardia remodeling links the potential mechanisms of atrial fibrillation, since atrial fibrillation beginning by any mechanism is likely to cause tachycardia-remodeling and thus promote the maintenance of atrial fibrillation by multiple-circuit reentry. Atrial structural remodeling is discussed as a paradigm of atrial fibrillation in which the classic features required for reentry (reduced refractory period and reentrant wavelength) may be lacking. Finally, the importance of recent insights into potential genetic determinants of atrial fibrillation is reviewed. The classic understanding of atrial fibrillation pathophysiology saw the different possible mechanisms as being alternative and opposing hypotheses. We now consider the multiple potential mechanisms as contributing to the pathophysiology of the arrhythmia to a different extent in different clinical settings and interacting with each other in a dynamic way at various stages of the natural history in many patients. It is hoped that this improved mechanistic understanding will lead to the development of improved therapeutic options. PMID- 12746748 TI - Atrial fibrillation: epidemiologic considerations and rationale for conversion and maintenance of sinus rhythm. AB - Atrial fibrillation is now the most common cardiac arrhythmia for which a patient is hospitalized. Clinically, it presents in a form that is paroxysmal, persistent, or permanent and may be symptomatic or asymptomatic, occurring in the setting of either no cardiac disease ("lone atrial fibrillation") or, most often, in association with an underlying disease. Atrial fibrillation is associated with a 2-fold increase in mortality and, in the United States alone, causes over 75,000 cases of stroke per year. The annual prevalence of stroke is 5% to 7%, but the use of adequate anticoagulation can reduce this to less than 1%. Atrial fibrillation is a disorder of the elderly, with almost equal prevalence in men and women. In the United States, 80% of atrial fibrillation occurs in patients over the age of 65 years, and its prevalence tracks that of heart failure, which may be the cause, as well as the result, of the arrhythmia. Both conditions are increasing in epidemic proportions in the aging population. The most common causes of atrial fibrillation are hypertensive heart disease, coronary artery disease, and heart failure with a miscellany of lesser conditions, with about 10% lacking structural heart disease. Unlike other supraventricular arrhythmias, cure by the use of catheter ablation and surgical techniques has not been a reality except in a relatively small number of cases. However, restoration and maintenance of sinus rhythm remain the initial goal of therapy for most patients. Pharmacologic approaches remain the mainstay of therapy for rate control and anticoagulation as well as for maintenance of sinus rhythm following pharmacological or electrical conversion. The changing epidemiology of atrial fibrillation is highlighted, with the focus on its conversion by the use of newer and novel antifibrillatory agents relative to the mechanisms of the arrhythmia, to restore the stability of sinus rhythm. PMID- 12746749 TI - Antiarrhythmic drug therapy of atrial fibrillation: focus on new agents. AB - The precise mechanisms of clinical effect of antiarrhythmic agents and the ideal "molecular targets" against arrhythmias, in particular atrial fibrillation, are poorly understood. Current antiarrhythmic drug development, particularly for drugs expected to be active against atrial fibrillation, has focused on drugs with multiple ionic mechanisms of action, in particular on those that block multiple potassium channels. Investigation of antiarrhythmic agents is complicated by the diversity of animal-disease models studied, by the potential multiple mechanisms of arrhythmias, and by the incompletely understood relationships between risks and benefits of antiarrhythmic drug therapy. Furthermore, rhythm control strategies in large groups of patients with atrial fibrillation have failed to show substantial clinical benefit. Nevertheless, drugs that block multiple potassium channels and appear to have relatively little organ toxicity, such as tedisamil, may represent an important new avenue in the therapeutic approach to highly symptomatic arrhythmias such as atrial fibrillation. PMID- 12746750 TI - Tedisamil in coronary disease: additional benefits in the therapy of atrial fibrillation? AB - Atrial fibrillation has recently come into clinical and research focus. In particular, ventricular rate control has been carefully compared with atrial rhythm control. Additionally, the recent discovery of atrial stunning has initiated clinical and research interest in atrial remodeling. Atrial fibrillation is more likely to occur when the atria are damaged by increased fibrosis. The ideal way to prevent atrial fibrillation and the risk of repetition is by tackling the root causes, such as ischemic heart disease, heart failure, and left ventricular hypertrophy. Tedisamil is an unusual antifibrillatory compound that has a novel mechanism of action by inhibiting the transient outward current (Ito) and the repolarizing potassium currents in the sinoatrial node. Tedisamil works acutely against atrial fibrillation. Importantly, atrial fibrillation is often caused by or related to cardiac ischemia, and conversely, ischemia is caused by the increased oxygen demand of atrial fibrillation. Hence, the double properties of tedisamil as a drug that both inhibits atrial fibrillation and acts in an anti-ischemic mode are an attractive basis for future clinical research. PMID- 12746751 TI - Rhythm or rate control in atrial fibrillation: insights from the randomized controlled trials. AB - Pharmacologic treatment remains the mainstay of therapy in patients with atrial fibrillation for the maintenance of normal sinus rhythm. Initial therapy of atrial fibrillation is often directed toward the maintenance of sinus rhythm by means of cardioversion and the use of antiarrhythmic drugs. Heart rate control is often only pursued when rhythm control fails. Four randomized controlled trials have carefully evaluated the yield of these two treatment strategies as the initial approach to patients with paroxysmal or persistent atrial fibrillation. In essence, all four trials demonstrated that an initial strategy of rate control is equally effective compared to the rhythm control approach in terms of clinically important outcome measures including mortality, stroke prevention, or quality of life. Accordingly, rate control can be considered as an initial approach to therapy in patients with paroxysmal or persistent atrial fibrillation. The four randomized trials clearly demonstrate that continuous anticoagulation is mandatory in all patients with atrial fibrillation and risk factors for stroke, irrespective of the initial therapeutic approach of rhythm or rate control. PMID- 12746753 TI - Androgen-induced vasorelaxation: a potential vascular protective effect. AB - The incidence of morbidity and mortality due to cardiovascular disease is well known to be higher in men than in women. For many years, experimental and clinical studies have focused on the role of androgens in cardiovascular pathology. The clinical implications of androgen deficiency as well as the molecular mechanism of their action have not been well established yet. However, an increasing amount of evidence suggests that androgens may have beneficial effects on the cardiovascular system. The purpose of this article is to review the evidence regarding the vasodilating effect induced by androgens, the mechanism of their action as well as their structure-function relationship. PMID- 12746754 TI - Fat modification in the diabetes diet. AB - The modification of dietary fat in the diet of diabetic patients is of interest with respect to metabolic and other consequences of this modification. To begin with the data are reviewed for the use of monounsaturated fatty acids (MUFA) in the diabetes diet. Compared to a carbohydrate-rich diet, glucose concentrations are lower. Blood pressure was also found to be lower. There were no major differences with respect to lipid concentrations. HDL-cholesterol levels tended to be higher after a MUFA-rich diet. In type-1 diabetic patients, the number of circulating big VLDL particles was greater after a MUFA diet than after a carbohydrate-rich diet. Comparisons were also made between diets enriched with MUFA and with polyunsaturated fatty acids (PUFA). With respect to lipid concentrations, different groups observed different effects. While one group saw no differences in fasting lipids, they measured a higher remnant-like particle cholesterol after a diet enriched with MUFA. Another group found higher total and LDL-cholesterol levels after a PUFA-rich diet than after a MUFA-diet. In their study, fasting glucose, insulin and fasting chylomicrons and postprandial chylomicrons and VLDL were higher following the PUFA diet. A MUFA-rich diet increased endothelium-dependent flow-mediated dilatation in the superficial femoral artery. Alpha-linolenic acid appears to be a precursor of eicospentaenoic and docosahexaenoic fatty acids. As a diet rich in n-6 PUFA reduces this conversion, a n-6/n-3 PUFA ratio not exceeding 4 - 6 should be observed. No prospective data are available for alpha-linolenic acid in diabetic patients. The review summarizes the results of the Lyon Diet Heart Study and the Nurses' Health Study. Both studies saw a reduced cardiovascular risk associated with a higher intake of alpha-linolenic acid. Finally, data on the effects of fish oil are given. The latter has a clearly expressed triglyceride-lowering effect. Data with respect to glucose control are heterogeneous. Major studies did not find any influence in glucose concentrations. Hepatic glucose production and peripheral insulin sensitivity remained constant. Evidently, nerve function can be improved by fish oil. Data have been compiled comparing the effects of fish oil with those of olive oil, linseed oil and sunflower oil. PMID- 12746755 TI - The effects of triiodothyronine on human osteoblast-like cells metabolism and interactions with growth hormone. AB - The expression of thyroid hormones receptors in osteoblasts and osteoclasts has involved these cells as direct targets for triiodothyronine (T3), but thyroid hormones may also interact with other hormones or local growth factors to exert their actions on bone cells. Among these, growth hormone (GH) is recognised as participating in the acquisition and maintenance of bone mass and exerting stimulatory effects on human osteoblastic cells. The aim of this study was to investigate T3 effects on primary human adult osteoblast-like cells (HOB) as well as to test for possible interactions between T3 and GH on bone cell metabolism. Primary human bone cell cultures were obtained by outgrowth from trabecular bone fragments from the hip and knee. Dose-response studies demonstrated enhanced [3H] thymidine incorporation for T3 at 10(-9), 10(-8), 10(-7) and 20(-7) M, with a maximal response of 162.81 +/- 12.97 % with T3 10(-8) M, compared to vehicle (p < 0.001). Time-course studies showed an increased osteoblast-like cell proliferation after 24 h, followed by a decrease of cell proliferation by 48 h and 72 h of culture, respectively, when compared to control cells, with a maximal response after 72 h (T3 10(-10) M: 45.21 +/- 6.97 %, p < 0.01). In addition, T3 markedly increased specific alkaline phosphatase (AP) activity in HOB (10(-10) M: 169.86 +/- 12.14 % vs. control, p < 0.001), but no significant influence on type I procollagen propeptide (PICP) production was observed. At 10(-9) - 10(-7) g/ml, GH significantly enhanced HOB proliferation (p < 0.001) however, GH effects were not dose-dependent. Triiodothyronine, at a high concentration (10(-7) M), stimulated GH-receptor (GHR) mRNA levels by 165.20 +/- 16.54 % after 24 h (p < 0.05). Correspondingly, a synergistic effect of T 3 with the same concentration and GH on cell proliferation in human adult osteoblast-like cells was found. PMID- 12746756 TI - No evidence for involvement of the promoter polymorphism -866 G/A of the UCP2 gene in childhood-onset obesity in humans. AB - Recently, an association between obesity and the G-allele of the - 866 G/A polymorphism in the promoter region of uncoupling protein-2 gene (UCP2) was reported. Both allele frequencies and genotype distributions for this polymorphism differed between obese individuals and never-obese controls. We attempted to confirm this finding. Genotyping was performed by polymerase chain reaction with subsequent restriction fragment length polymorphism analysis (PCR RFLP). We analysed transmission disequilibrium of the (wild type) G-allele for 200 extremely obese children and adolescents from 93 concordant sib pair families using the pedigree disequilibrium test. Additionally, using a one-sided asymptotic Pearson's chi 2-test, we tested whether the G-allele occurs more frequently in 277 extremely obese children and adolescents (including the 93 index patients of the concordant sib pairs) than in 188 never-obese controls. The one-sided asymptotic Cochran Armitage trend test was used to determine differences in genotype frequencies between extremely obese and healthy underweight individuals. The PDT analysis revealed no evidence for transmission disequilibrium in obesity. Allele and genotype frequencies did not differ between the extremely obese and never-obese subjects. In conclusion, we cannot confirm the results of ) in our young sample. PMID- 12746757 TI - Chronic treatment with inhaled corticosteroids does not modify leptin serum levels. AB - AIM: To assess the influence of a short-term treatment with low-dose inhaled corticosteroids on leptin serum levels. PATIENTS: 14 prepubertal children, mean age 5.1 +/- 2.4 years, treated with inhaled fluticasone propionate 100 microg b.d. and 16 prepubertal children, mean age 8.3 +/- 1.3 years, treated with inhaled budesonide 200 microg b.d. METHODS: All children underwent a CRH test with evaluation of leptin, cortisol and ACTH levels before and after 3 months of treatment. RESULTS: Fluticasone group: no difference was found between basal cortisol level, delta and area under the curve (AUC) before and after treatment, though cortisol peak was significantly lower following treatment. Basal ACTH level, peak and AUC were significantly lower after treatment. Budesonide group: no statistically significant difference in any of the parameters regarding cortisol and ACTH secretion was observed before and after treatment. No significant changes in basal serum leptin levels and AUC were observed following treatment in both groups. Furthermore no significant variation in leptin level was observed during both CRH tests. DISCUSSION: Leptin secretion does not seem to be affected by low-dose inhaled corticosteroids; moreover leptin does not seem to be involved in the response of the HPA axis to stress. PMID- 12746758 TI - Endothelin-1 levels in patients with disorders of the thyroid gland. AB - The endothelium derived peptide endothelin-1 (ET-1) is the major isoform of the endothelin peptide family, which is produced and secreted in the endothelial cell system. We measured plasma levels in patients with thyroid diseases and investigated associations between laboratory and clinical markers of thyroid metabolism and ET-1 plasma levels. ET-1 plasma levels were determined in patients with Graves' disease (n = 54), endemic goiter (n = 26), patients with Hashimoto's thyroiditis (n = 21) and compared to healthy controls (n = 60). ET-1 plasma levels were significantly elevated in patients with Hashimoto's thyroiditis (p < 0.0001) and in patients with Graves' disease (p = 0.003), when compared to healthy controls. In patients with endemic goiter, no significant differences were found compared to healthy controls (p = 0.298) and when compared to patients with Graves' disease (p = 0.16). We did not observe an association between ET-1 plasma levels and parameters of thyroid disease (e.g. thyroidea-stimulating hormone, thyroxine, volume of the thyroid). Furthermore, patients with and without endocrine thyroid disease showed no significantly different ET-1 plasma levels (p = 0.78). These data suggest that the autoimmunologically induced inflammatory response of the thyroid gland in Hashimoto's thyroiditis and Graves' disease is responsible for increased ET-1 plasma levels. Furthermore, our data do not support a role for ET-1 as a valid quantitative indicator for stage or progression in endemic goiter, Graves' disease or Hashimoto's thyroiditis. PMID- 12746759 TI - Analysis of the relationship between PPAR-gamma 2 gene variants and severe insulin resistance in obese patients with impaired glucose tolerance. AB - Mutations in the peroxisome proliferator-activated receptor-gamma 2 (PPAR-gamma 2) gene may cause obesity and insulin resistance. Therefore we investigated whether known variants in the PPAR-gamma 2 gene are associated with obesity and extreme insulin resistance in obese patients with impaired glucose tolerance (IGT). The Pro115 Gln, Pro12Ala, Pro467Leu, Val290Met and a silent polymorphism C478 T were examined in 48 subjects with IGT and insulin resistance (IR), characterized by euglycemic hyperinsulinemic clamps, and in 52 healthy insulin sensitive (IS) controls. We found one proband in the IR group with the Pro115 Gln variant. This subject showed a lower whole body glucose uptake (18 micromol/kg per min) compared to the entire IR group (29 micromol/kg per min). The body weight of the proband (BMI 28.5 kg/m2) was within the average of the IR group (30.3 +/- 0.8 kg/m2). The Pro12Ala variant was not associated with differences in BMI, in the degree of insulin resistance between the IR and IS group. The Pro467Leu, Val290Met mutations and the silent polymorphism CAC478CAT were not detected in any group. In conclusion, the Pro115 Gln variant, but not the Pro12Ala mutation in the PPAR-gamma 2 gene, could be a rare cause of severe insulin resistance. PMID- 12746760 TI - Pioglitazone increases serum magnesium levels in glucose-intolerant subjects. A randomized, controlled trial. AB - Although thiazolidinediones and magnesium supplementation improves insulin action and increases HDL-cholesterol, the potential link between serum magnesium and thiazolidinediones has received little attention. Focusing on the increase of serum magnesium, 63 eligible subjects were enrolled and randomly allocated to receive either 30 mg Pioglitazone once daily (Group A) or lifestyle intervention (Group B) during 12 weeks. Subjects were eligible if they were glucose intolerant, and excluded if they had high blood pressure, diabetes or abnormal liver function tests. The personnel assessing outcomes were blinded to group assignment. Of the 63 eligible subjects, 3 dropped out (one in group A, and two in Group B) because they moved out of the city. So, 30 subjects in each group, who satisfactorily completed the follow-up, were included in the analysis of data. There were no serious adverse events or side effects due to Pioglitazone or lifestyle intervention. At baseline, the groups did not differ significantly in serum magnesium levels 1.73 +/- 0.17 versus 1.72 +/- 0.14 mg/dl, p = 0.80. Subjects who received Pioglitazone significantly increased their serum magnesium to 1.93 +/- 0.16 mg/dl whereas in the lifestyle intervention group the increase was 1.74 +/- 0.25 mg/dl, p < 0.0001. What this study showed was a significant increase in the serum magnesium levels of glucose-intolerant subjects who received 30 mg Pioglitazone once daily. PMID- 12746761 TI - Metabolic effects of mealtime insulin lispro in comparison to glibenclamide in early type 2 diabetes. AB - The efficacy and safety of the preprandial injection of insulin lispro was compared with the oral administration of glibenclamide in patients with early type 2 diabetes. In this open-label, multicenter study, 143 patients with a glucagon-stimulated increase in C-peptide of at least 0.4 nmol/L were randomized to receive preprandial insulin lispro (LP) or glibenclamide (GB) for 26 weeks. Seventy-five patients received LP (51 male/24 female; age 40 to 70 years, duration of diabetes 4.4 +/- 2.9 years) and 68 patients received GB (39 male/29 female; age 39 to 70 years; duration of diabetes 4.3 +/- 3.4 years). After 12 weeks, mean 90 minute blood glucose excursions were 0.9 +/- 1.0 mmol/L for LP and 1.8 +/- 1.2 mmol/L for GB (p < 0.0001). After 24 weeks, mean blood glucose excursions were 1.0 +/- 1.1 mmol/L for LP and 1.7 +/- 1.2 mmol/L for GB (p = 0.002). Body weight decreased slightly from 87.2 +/- 2.3 to 86.5 +/- 12.2 kg in the LP group and increased from 84.1 +/- 13.7 to 84.4 +/- 13.3 kg in the GB group. LP versus GB induced changes from baseline to endpoint in fasting C peptide (nmol/L), proinsulin and insulin levels (pmol/L) were - 0.2 +/- 0.4 versus - 0.1 +/- 0.6 (p = 0.04), - 11.2 +/- 26.0 versus - 1.1 +/- 17.3 (p = 0.03), and - 27.8 +/- 147.4 versus + 32.6 +/- 286.2 (not significant), respectively. HbA 1c at baseline was 7.5 +/- 1.0 % for LP and 7.7 +/- 1.2 % for GB and did not change significantly in either group during the investigation. No significant difference was observed between the groups with respect to hypoglycemic episodes. Treatment with LP improved postprandial blood glucose control more than GB without increasing body weight or hypoglycemic episodes. In addition, use of LP was associated with a decrease in fasting C-peptide and proinsulin levels, suggesting a potential down regulation of endogenous insulin production and improved proinsulin processing efficiency. PMID- 12746762 TI - Impairment of cutaneous arteriolar 0.1 Hz vasomotion in diabetes. AB - Arteriolar vasomotion, the cyclic contraction/dilation of terminal arterioles, is disordered in diabetes. The aim of the present study was to characterize the impairment of cutaneous vasomotion in type 1 and type 2 diabetes, especially with regard to the influence of metabolic control and to the response to shear stress. Twenty type 1 and 23 type 2 diabetic patients were investigated. Vasomotion waves were recorded in single capillaries at the dorsal middle phalangeal area of the left ring finger during rest, after warming the skin temperature to 33 degrees C, and after 3-min arterial occlusion by means of laser Doppler anemometry. Suprasystolic occlusion caused an increase in amplitudes of vasomotion only in type 1 diabetic patients (0.12 +/- 0.04 mm/s vs. 0.36 +/- 0.06 mm/s, p = 0.001). In type 1 but not in type 2 diabetic patients, both systolic and diastolic blood pressure correlated positively with amplitudes of resting vasomotion (r = 0.62, p = 0.002 and r = 0.65, p = 0.001, respectively). Amplitudes of vasomotion after warming up at frequencies of 5 - 8 cycles per minute (0.08 - 0.13 Hz) correlated inversely with the levels of glycated hemoglobin (HbA 1c ) (r = - 0.56, p = 0.005) only in type 1 diabetic patients. In conclusion, we found suprasystolic occlusion and increasing blood pressure to provoke vasomotion with a concomitant decrease in effective vascular resistance only in type 1 diabetic patients. The impaired vasomotion response to shear stress in type 2 diabetes might favour the development of skin lesions and arterial hypertension. Insufficient glycemic control seems to be an important factor in the pathogenesis of impaired vasomotion in type 1 diabetes. PMID- 12746763 TI - Adrenal cortical phaeochromocytoma: a case report of a rare entity. AB - Adrenal cortical phaeochromocytomas (pseudo-phaeochromocytomas) are a very rare entity and a diagnostic challenge. Of the few cases previously reported, most have incomplete data or lack clinical and biochemical follow-up documenting the cure of the excess secretion of catecholamines after resection of the tumour. We report herein a 62-year-old patient with clinical and biochemical findings diagnostic of a phaeochromocytoma associated with a 2-cm adrenal mass on CT scan. Surgery revealed the presence of an adrenal cortical adenoma with positive staining for the neuroendocrine marker synaptophysin, but negative for chromogranin, as has been previously reported for these rare cortical phaeochromocytomas. After removal of the tumour the clinical symptoms resolved and biochemical markers normalized, demonstrating the causal relationship between the cortical tumour and the excess production of catecholamines. PMID- 12746764 TI - Triad of acute infusion-related reactions associated with liposomal amphotericin B: analysis of clinical and epidemiological characteristics. AB - We investigated the clinical characteristics and treatment of patients with a distinctive triad of acute infusion-related reactions (AIRRs) to liposomal amphotericin B (L-AMB) via single-center and multicenter analyses. AIRRs occurred alone or in combination within 1 of 3 symptom complexes: (1) chest pain, dyspnea, and hypoxia; (2) severe abdomen, flank, or leg pain; and (3) flushing and urticaria. The frequency of AIRRs in the single-center analysis increased over time. Most AIRRs (86%) occurred within the first 5 min of infusion. All patients experienced rapid resolution of symptoms after intravenous diphenhydramine was administered. The multicenter analysis demonstrated a mean overall frequency of 20% (range, 0%-100%) of AIRRs among 64 centers. A triad of severe AIRRs to L-AMB may occur in some centers; most of these reactions may be effectively managed by diphenhydramine administration and interruption of L-AMB infusion. PMID- 12746766 TI - Fluconazole plus amphotericin B combinations are not contraindicated and may add benefit for the treatment of candidemia. PMID- 12746767 TI - Efficacy of a targeted, oral penicillin-based yaws control program among children living in rural South America. AB - Yaws is endemic in rural Guyana. An observational study was conducted to determine the efficacy of oral penicillin V therapy in treating skin lesions of yaws in children. In 1999, inhabitants of 7 rural villages near Bartica, Guyana, were screened for skin lesions of yaws. Cases were confirmed by serological testing. A control program was implemented in 2000: children < or =14 years old were screened, and those with active lesions were treated with oral penicillin V for 7-10 days. In 2001, children were rescreened and active cases were treated. Prevalence of yaws skin lesions fell from 5.1% (52 of 1020 children screened in 2000) to 1.6% (8 of 516 in 2001), a 71% drop. Sixteen (94%) of 17 children treated in 2000 and reassessed in 2001 had complete resolution of lesions. A targeted, oral penicillin-based treatment regimen can successfully treat dermatologic yaws in individual children and can decrease the prevalence of skin yaws in a community in which it is endemic. This information may aid in the implementation of additional control efforts. PMID- 12746765 TI - A randomized and blinded multicenter trial of high-dose fluconazole plus placebo versus fluconazole plus amphotericin B as therapy for candidemia and its consequences in nonneutropenic subjects. AB - A randomized, blinded, multicenter trial was conducted to compare fluconazole (800 mg per day) plus placebo with fluconazole plus amphotericin B (AmB) deoxycholate (0.7 mg/kg per day, with the placebo/AmB component given only for the first 5-6 days) as therapy for candidemia due to species other than Candida krusei in adults without neutropenia. A total of 219 patients met criteria for a modified intent-to-treat analysis. The groups were similar except that those who were treated with fluconazole plus placebo had a higher mean (+/- standard error) Acute Physiology and Chronic Health Evaluation II score (16.8+/-0.6 vs. 15.0+/ 0.7; P=.039). Success rates on study day 30 by Kaplan-Meier time-to-failure analysis were 57% for fluconazole plus placebo and 69% for fluconazole plus AmB (P=.08). Overall success rates were 56% (60 of 107 patients) and 69% (77 of 112 patients; P=.043), respectively; the bloodstream infection failed to clear in 17% and 6% of subjects, respectively (P=.02). In nonneutropenic subjects, the combination of fluconazole plus AmB was not antagonistic compared with fluconazole alone, and the combination trended toward improved success and more rapid clearance from the bloodstream. PMID- 12746768 TI - Is azithromycin the first-choice macrolide for treatment of community-acquired pneumonia? AB - Combination treatment with a beta-lactam plus a macrolide may improve the outcome for elderly patients with community-acquired pneumonia (CAP). The prognoses and mortality rates for elderly patients with CAP who receive ceftriaxone combined with a 3-day course of azithromycin or a 10-day course of clarithromycin were compared in an open-label, prospective study. Of 896 assessable patients, 220 received clarithromycin and 383 received azithromycin. There were no significant differences between groups with regard to the severity score defined by the Pneumonia Patient Outcomes Research Team (PORT) study group; the incidence of bacteremia was also not significantly different. However, for patients treated with azithromycin, the length of hospital stay was shorter (mean+/-SD, 7.4+/-5 vs. 9.4+/-7 days; P<.01) and the mortality rate was lower (3.6% vs. 7.2%; P<.05), compared with those treated with clarithromycin. There might be a difference in the outcome for patients with CAP depending on the macrolide used. A shorter treatment course with azithromycin may result in better compliance with therapy. PMID- 12746769 TI - Mycoplasma hominis lipid-associated membrane protein antigens for effective detection of M. hominis-specific antibodies in humans. AB - Lipid-associated membrane proteins (LAMPs) from 14 Mycoplasma hominis isolates or strains share similar protein and antigenicity profiles. Of 31 human immunodeficiency virus-infected patients from whose samples M. hominis was cultured, 28 tested strongly positive for serum antibodies to M. hominis LAMPs. The remaining 3 serum samples showed low antibody titer to LAMPs from all of the 14 M. hominis isolates or strains, which was likely the result of the compromised immune systems of the patients. Thus, M. hominis LAMPs as a whole are homogenous in antigenicity within the species, despite having many different serotypes. Serological study involving 564 healthy blood donors and 211 patients attending sexually transmitted disease clinics by LAMPs showed that general populations were widely exposed to M. hominis. Women were infected with M. hominis at a younger age than were men. The prevalence of infection increased markedly among sexually active persons. PMID- 12746770 TI - Imported Lassa fever in Germany: surveillance and management of contact persons. AB - This study sought to assess the risk of secondary transmission after import of Lassa fever into Europe. A total of 232 persons exposed to a case of Lassa fever imported into Germany were identified. The level of exposure was determined for 157 persons (68%), and 149 (64%) were tested serologically. High-risk or close contact was reported by 30 (19%) of 157 persons. No symptomatic secondary infections were observed. However, Lassa virus-specific immunoglobulin G antibodies were detected in a serum sample obtained from a physician who examined the index patient on day 9 of illness. The physician received ribavirin prophylaxis and did not develop symptoms of Lassa fever. On the basis of these data, the contact was classified as having a probable secondary infection. The study indicates a low risk of transmission during the initial phase of symptomatic Lassa fever, even with high-risk exposures. The risk may increase with progression of disease and increasing virus load. PMID- 12746771 TI - Inflammation and coagulation: implications for the septic patient. AB - Sepsis with acute organ dysfunction (severe sepsis) is common, frequently fatal, and associated with a significant national health/economic burden. In addition to standard care, investigators have focused on interrupting the inflammatory and anti-inflammatory cascade associated with this disease. Unfortunately, despite promising preclinical results, interventions directed at the inflammatory elements have not reduced the morbidity and mortality associated with this disease. Inflammation and coagulation are tightly linked. In fact, sepsis associated coagulopathy is almost universal in patients with severe sepsis. Preclinical observations indicate that antithrombotic-targeted therapy has the potential to reduce morbidity and mortality in patients with this disease. Treatment with recombinant human activated protein C (drotrecogin alpha [activated]) was the first antithrombotic-targeted therapy to significantly reduce 28-day all-cause mortality in patients with severe sepsis. The pathophysiological and clinical significance of this evidence and the relationship of coagulation to inflammation are discussed, as are positive and negative results of clinical trials of antithrombotic therapy. PMID- 12746772 TI - Considerations in control and treatment of nosocomial infections due to multidrug resistant Acinetobacter baumannii. AB - We sought to control infection due to multidrug-resistant Acinetobacter baumannii (MDR-Ab) by identifying isolates as clonally related, leading to enhanced infection-control measures, including cohorting, surveillance, contact precaution, initial therapy with ampicillin/sulbactam and local polymyxin B, and, more recently, therapy with synergistic antibiotic combinations. Class restriction of cephalosporins has been associated with a reduction in cephalosporins-cephamycin-carbapenem resistance among nosocomial Klebsiella isolates. This has been supplemented by restriction of carbapenem use after an initial 24-h period in an effort to reduce the selection of porin-deficient, carbapenem-resistant A. baumannii and Pseudomonas aeruginosa. Evidence is reviewed suggesting that eradication of MDR-Ab nosocomial colonization may prevent subsequent infection. Relatively few standard antibacterial drugs remain active against MDR-Ab. Published clinical results of therapy with these agents are reviewed, and in vitro evidence of synergy between them is presented that suggests that combination therapy should be studied for enhanced clinical activity. PMID- 12746773 TI - Inhalational anthrax due to bioterrorism: would current Centers for Disease Control and Prevention guidelines have identified the 11 patients with inhalational anthrax from October through November 2001? AB - A panel of 10 physicians used the nominal group technique to assess the ability of the Centers for Disease Control and Prevention (CDC) interim guidelines for clinical evaluation of persons with possible inhalational anthrax (IA) to retrospectively identify the 11 patients with IA seen during the October 2001 bioterrorism outbreak. The guidelines would not have identified 10 of 11 of these patients, primarily because the guidelines were designed to address only those patients with a known history of exposure or clearly identified environmental or occupational risk. The panel suggested revisions to the guidelines, primarily consisting of broadening the criteria for evaluation to include either known exposure or environmental occupational risk, or to include clinical symptoms consistent with IA. These extensions of the guidelines retrospectively identified 8 of 11 of the patients with IA from October 2001. PMID- 12746774 TI - Antimicrobial approaches for preventing infections associated with surgical implants. AB - Because management of infections associated with surgical implants can be both difficult and costly, prevention of such infections remains a priority. Preventive strategies comprise systemic perioperative administration of antibiotics and local application of antimicrobial agents (antibiotics or antiseptics). Local antimicrobial prophylaxis can be provided in various forms and aims to prevent implant-associated infections by impeding bacterial adherence to the implant surface and/or reducing the concentration of bacteria in the immediate vicinity of the implant. Analysis of the existing clinical practices and the pertinent medical literature indicates that, although some antimicrobial strategies constitute the standard of care for preventing infections associated with surgical implants, such strategies are often applied in a nonstandardized fashion and without clear evidence of clinical efficacy. This review article concludes with a perspective on assessing and preventing such serious infections. PMID- 12746775 TI - Topical microbicides for disease prevention: status and challenges. AB - As the human immunodeficiency virus and sexually transmitted disease epidemics advance relentlessly, it is clear that an armamentarium of therapeutic and preventive methods will be essential to their containment. Topical microbicides- self-administered prophylactic agents applied to the vagina or rectum in various formulations--could be a crucial addition to that necessary armamentarium. This article provides an update on the dramatically broadening array of approaches being pursued in microbicide research and development and identifies critical challenges to progress. PMID- 12746777 TI - Outbreak of tuberculosis among homeless persons coinfected with human immunodeficiency virus. AB - We investigated a cluster of patients with tuberculosis (TB) in North Carolina and determined the extent of transmission of 1 strain of Mycobacterium tuberculosis. A retrospective cohort study was conducted. Homeless shelter attendance and medical records for 1999 and 2000 were reviewed. The period of exposure to M. tuberculosis was determined, and shelter residents were offered TB screening. DNA fingerprinting was performed on 72 M. tuberculosis isolates. In addition to the initial index cluster of 9 patients, another 16 patients were identified. Isolates of M. tuberculosis from all 25 patients shared a matching DNA fingerprint pattern. All but 1 patient was male, 22 (88%) were African American, and 14 (56%) were human immunodeficiency virus-infected. An epidemiological link to a single shelter was identified for all but 1 patient. Earlier recognition of this shelter as a site of M. tuberculosis transmission could have been facilitated through innovative approaches to contact investigation and through genetic typing of isolates. PMID- 12746776 TI - Interferon-ribavirin in association with stavudine has no impact on plasma human immunodeficiency virus (HIV) type 1 level in patients coinfected with HIV and hepatitis C virus: a CORIST-ANRS HC1 trial. AB - A randomized, open-label trial was performed to study virological and intracellular interactions between stavudine and ribavirin in 30 patients coinfected with human immunodeficiency virus (HIV) and hepatitis C virus (HCV). Patients were randomized to receive either interferon and ribavirin or no treatment for HCV infection for 3 months. Intracellular peripheral blood mononuclear cells' stavudine-triphosphate (TP) concentrations were assessed. Plasma HIV RNA levels did not change significantly between baseline and month 3. There was a nonstatistically significant trend for a lower median residual concentration of intracellular stavudine-TP in the treated group, compared with the control group. The same trend was also observed for peak concentrations. Coprescription of ribavirin and stavudine has no short-term impact on plasma HIV RNA level in HIV-HCV-coinfected patients treated with stavudine as a part of their antiretroviral treatment; this coprescription can be safely used, although an in vivo interaction between ribavirin and stavudine is possible. PMID- 12746778 TI - Prevalence and characteristics of hepatitis C virus coinfection in a human immunodeficiency virus clinical trials group: the Terry Beirn Community Programs for Clinical Research on AIDS. AB - The baseline prevalence of hepatitis C virus (HCV) and human immunodeficiency virus (HIV) coinfection among 2705 patients enrolled in HIV clinical trials in the Community Programs for Clinical Research on AIDS (CPCRA) was 16.6%. For men, multivariate logistic regression showed that the baseline prevalence of HIV-HCV coinfection was positively associated with history of injection drug use, older age, antiretroviral therapy naive status, African American or Latino ethnicity, and no history of having sex with men. No association was found with baseline CD4+ cell count or HIV RNA level. The prevalence of HCV coinfection in a diverse HIV clinical trials cohort provides additional information about risk behaviors and demographic factors that can be used in the analysis of clinical and virologic outcomes. PMID- 12746779 TI - Association of hepatitis C infection and antiretroviral use with diabetes mellitus in drug users. AB - We performed a cross-sectional analysis of factors associated with diabetes mellitus among 557 drug users who were recruited from a methadone treatment program and were participating in a longitudinal study of hepatitis C virus (HCV) infection. We found that HCV infection was strongly associated with diabetes mellitus (adjusted odds ratio [OR], 2.9; 95% confidence interval [CI], 1.3-6.4) after controlling for age, race, unemployment, and body mass index. Among HIV infected drug users, receipt of antiretroviral therapy (ART) for >1 year was also associated with diabetes (adjusted OR for receipt of ART without a protease inhibitor, 4.1 [95% CI, 1.1-15.5]; adjusted OR for receipt of ART including a protease inhibitor, 5.5 [95% CI, 1.5-20.4]). PMID- 12746781 TI - Discontinuation of secondary prophylaxis for cryptococcal meningitis in human immunodeficiency virus-infected patients treated with highly active antiretroviral therapy: a prospective, multicenter, randomized study. AB - A prospective, multicenter, randomized study was conducted with human immunodeficiency virus (HIV)-infected patients who were successfully treated for acute cryptococcal meningitis, were receiving secondary prophylaxis, and were naive for antiretroviral therapy. Patients were randomized to continue or discontinue secondary prophylaxis when the CD4 cell count had increased to >100 cells/microL and an undetectable HIV RNA level had been sustained for 3 months. At a median of 48 weeks after randomization, there were no episodes of cryptococcal meningitis in either group. PMID- 12746780 TI - Risk factors for lactic acidosis in HIV-infected patients treated with nucleoside reverse-transcriptase inhibitors: a case-control study. AB - A case-control study was undertaken to determine risk factors for lactic acidosis in human immunodeficiency virus-infected patients treated with nucleoside reverse transcriptase inhibitors (NRTIs). From May 1996 to June 2000, 9 patients with lactic acidosis (defined as a plasma lactic acid level of >5 mM and plasma pH of <7.38) were identified. Control patients were randomly selected from among a large cohort of patients who initiated a dual NRTI regimen in 1996 or after. Two factors were associated with an increased risk of lactic acidosis: first, a creatinine clearance of <70 mL/min before lactic acidosis (OR, 15.8 [range, 3.0 86.5], P<10(-4)), and, second, a low nadir CD4+ T lymphocyte count before the inception of NRTI therapy (OR, 8.4 [range, 1.2-infinity], P=.03). The total cumulative exposure to NRTIs was not associated with an increased risk of lactic acidosis, nor was the cumulative exposure to any of the 4 NRTIs studied. According to these results, monitoring of creatinine clearance, especially in patients with a low nadir CD4+ T lymphocyte count, could lead to modifications in antiretroviral therapy in order to diminish the risk of occurrence of lactic acidosis. PMID- 12746782 TI - Limits of early diagnosis of herpes simplex encephalitis in children: a retrospective study of 38 cases. AB - The prognosis of herpes simplex encephalitis (HSE) depends on the early and appropriate administration of specific antiviral therapy. We retrospectively reviewed 38 cases of children with proven HSE, to evaluate the reliability of polymerase chain reaction results, according to the time of cerebrospinal fluid (CSF) sampling. Initial negative results were observed in 8 of 33 CSF samples drawn before day 3 of the disease and were significantly associated with a low level of protein and <10 leukocytes/mm3 in the CSF. PMID- 12746783 TI - Pneumocystis jiroveci genotypes and primary infection. AB - This study describes the molecular typing of Pneumocystis jiroveci organisms from 5 nonpremature immunocompetent infants who developed a primary infection. Four P. jiroveci internal transcribed spacer (ITS) types were identified. All have been previously described in reports concerning immunosuppressed adults with pneumocystosis. Present data suggest that identical types can be implicated either in first contact or in additional contacts between fungus and host and that both immunocompetent infants and immunocompromised patients may be part of a common human reservoir for the fungus. PMID- 12746784 TI - Anthrax during pregnancy: case reports and review. AB - We review, in detail, 2 cases of anthrax during pregnancy, its maternal and perinatal complications, and its management. Patient 1 was a 33-year-old woman at 32 weeks of gestation. She had a submandibular eschar; extensive edema on her face, neck, and upper thorax that inhibited respiratory function; and fever. The patient was treated with penicillin G and prednisolone after the diagnosis of anthrax. She recovered within 10 days but delivered a preterm baby. Patient 2 was a 29-year-old woman at 33 weeks of gestation. Her anthrax lesion was on her right elbow, and therapy consisted of procaine penicillin. She also delivered a preterm baby. These 2 cases show that anthrax during pregnancy can be successfully managed, but preterm delivery could be a complication. PMID- 12746785 TI - HIV infection in older patients. PMID- 12746786 TI - Staphylococcus lugdunensis breast abscess: is it real? PMID- 12746787 TI - Roseolovirus DNA in the colonic mucosa of HIV-seropositive patients with diarrhea. PMID- 12746788 TI - Resolution of fungemia due to Fusarium species in a patient with acute leukemia treated with caspofungin. PMID- 12746789 TI - Pythium insidiosum reidentified as Gymnascella hyalinospora. PMID- 12746791 TI - Prolonged perilesional edema after treatment of parenchymal neurocysticercosis: methotrexate as a corticosteroid-sparing agent. AB - Treatment of neurocysticercosis with larvicidal agents is commonly complicated by seizures and transient neurologic deficits as a result of the host immune response to dying cysts. We report a case in which treatment with high-dose praziquantel resulted in prolonged perilesional edema requiring use of corticosteroids and corticosteroid-sparing agents for >1 year, suggesting a role for methotrexate in the management of this condition. PMID- 12746790 TI - Contamination of bone marrow products with an actinomycete resembling Microbacterium species and reinfusion into autologous stem cell and bone marrow transplant recipients. AB - Bacterial contamination of bone marrow or peripheral blood stem cell transplant products typically occurs with skin flora or, rarely, gram-negative organisms. We describe a clonal outbreak of contamination in transplant products caused by contamination with an aerobic actinomycete that occurred at our institution during the summer of 2001. From 1 July through 12 September 2001, 73 peripheral blood or bone marrow stem cell products were obtained from 39 patients, and 34 products were found to be contaminated with the outbreak strain. Fourteen patients were reinfused with contaminated cells, and the outbreak strain was isolated from the blood cultures for one patient. Investigation revealed multiple potential sources for contamination during the product cryopreservation process. The outbreak of contamination was aborted upon modification of the cryopreservation process. PMID- 12746792 TI - Improvement of symptomatic human immunodeficiency virus-related lymphoid interstitial pneumonia in patients receiving highly active antiretroviral therapy. AB - To analyze the impact of highly active antiretroviral therapy on lymphoid interstitial pneumonia (LIP), we reviewed the medical files of 5 human immunodeficiency virus (HIV)-infected patients in whom LIP was diagnosed during 1996-2001 who had never previously received antiretroviral treatment. Patients were mildly immunosuppressed at the time of diagnosis of LIP but had high plasma HIV loads and marked circulating and pulmonary CD8 hyperlymphocytosis. All patients improved clinically, radiologically, and functionally; improvement was accompanied by a drastic reduction in the virus load and an increase in the CD4 lymphocyte count. PMID- 12746793 TI - Miller Fisher variant of Guillain-Barre syndrome associated with lactic acidosis and stavudine therapy. AB - We describe the first case of Miller Fisher syndrome (opthalmoplegia, ataxia, and areflexia) associated with lactic acidosis as an adverse effect of receipt of an antiretroviral regimen containing stavudine. We review this syndrome in the context of recent descriptions of neuromuscular toxicities attributed to nucleoside analogue reverse-transcriptase inhibitor-induced mitochondrial toxicity. PMID- 12746794 TI - [Fifty years from the discovery of DNA]. AB - Fifty years have passed from the description of the double helix of the DNA molecule. During these years very important scientific discoveries have been made in genetics and molecular biology. In addition, twenty years ago Mullis invented an important technique, the polymerase chain reaction, which is currently used for replication and amplification of the DNA. Well, this amazing technique has been used by the Human Genome Project to identify approximately 35,000 genes. We are now in the post-genomic era that will enable the achievement of important discoveries in life sciences. PMID- 12746795 TI - [Anderson-Fabry's disease: diagnostic problems, therapeutic relevance, and clinical experience in the treatment of the disease with enzyme replacement therapy in nephropathic patients]. AB - Aim of this study was to confirm the initial results of a clinical trial on the treatment of Fabry's disease carried out in 13 Italian Nephrology Units. Fabry's disease is a rare, X-linked inherited disease, characterized by a-galactosidase (a-GAL) deficiency, a lysosomial enzymatic activity that results in the accumulation of neutral glycosphingolipids in the endothelial cells of the whole body, and causes painful crises, acroparesthesiae, angiokeratomas, corneal and lens dystrophy, and progressive damage to kidneys, heart and central nervous system, as well as potentially leading to death. The present availability of the recombinant form of a-GAL allows us to prevent or stop the long-term complications of this disease. A clinical trial, generously supported by Genzyme, was started on February 2001. In this trial 20 patients affected by Fabry's disease were periodically treated with agalsidase-beta, the commercial form of the enzyme. The initial results of the trial have indicated that the drug is capable of reducing both the number and intensity of painful crises, improving the patient's sensation of well-being, thus suggesting that this therapeutic approach might theoretically increase life expectancy in these patients. PMID- 12746797 TI - [Effect of an ambulatory program devoted to chronic renal insufficiency on the reduction of mobidity and hospitalization among patients at the beginning of dialysis treatment]. AB - BACKGROUND: Late nephrological referral of end-stage renal disease (ESRD) patients is associated with increased risk of emergent dialysis start and poor complications control. However, the relative contribution of pre-dialysis care organization is unknown. METHODS: All 175 consecutive patients who started chronic dialysis for ESRD at our Institution from 1.1.99 to 30.6.02 were grouped as follows: referred ? 3 months before dialysis, (A, n=50); followed by non dedicated specialists (B, n=74) or by pre-dialysis educational program personnel (PEP, n=51). We examined the first six months of hospitalization, uraemic complications control, type of dialysis initiation, and first dialysis modality. RESULTS: There was no difference in baseline characteristics and comorbidities among groups. PEP patients had higher creatinine clearance, haemoglobin, calcemia and BMI at initiation. They also made greater use of ACE-inhibitors and were more likely to have a planned start and choose peritoneal dialysis. Emergent starts were 50% (A 100%, B 45%, PEP 4%, p<0.001). Mean pre-dialysis hospitalization (due to in-patient emergency dialysis onset for unplanned starts and planned for access insertion for elective out-patient starts) was shorter among PEP patients (7days-PEP, 17days-B, 30days-A). Logistic regression confirmed the predictive role of PEP for emergent start (AOR 0.03, 0.001 to 0.101, p<0.001) even excluding late referrals (AOR 0.1, 0.033 to 0.306, p<0.001), independently of baseline characteristics and comorbidities. CONCLUSIONS: Pre-dialysis follow-up by dedicated personnel was more effective than traditional specialist care in reducing morbidity and health care resources utilization in patients starting dialysis. PMID- 12746796 TI - [Genetic basis for malformation-associated uropathy and renal dysplasia]. AB - The wide association between urinary tract malformations and dysplastic kidneys, known as CAKUT (Congenital Anomalies of the Kidney and Urinary Tract), could be caused by a single disorder of the embryonic development of the kidney and urinary tract. These complex patterns of development are under genetic control. A positive family history strongly suggests a genetic origin of these conditions. Linkage studies show an extreme genetic heterogenicity and an important phenotypic and clinical variability of the same mutation. Some urinary tract malformations have been investigated in the context of clinical syndromes. The renal-coloboma syndrome is an autosomal dominant human disease, secondary to mutation of the PAX2 transcription factor, characterized by optic nerve coloboma, renal anomalies and vesicoureteral reflux. However, most of the urinary tract anomalies can occur in isolation. Studies have shown the association of hereditary hydronephrosis with HLA antigens on chromosome 6 and the association of VUR with the mutations in a locus of chromosome 1. The higher frequency and severity of some uropathies in the male gender may be explained by a linkage disequilibrium phenomenon or a X-linked transmission pattern. For example, the mutations in the AGTR2 gene on chromosome X were observed in animal models but not yet confirmed in human subjects. Finally, the ACE gene polymorphism is associated with a higher incidence of congenital hypo-dysplastic kidneys and represents a significant risk factor for the development of progressive renal damage. PMID- 12746798 TI - [The optimization of the treatment and timing of the intervention by a nephrologist in pre-dialytic chronic renal insufficiency]. AB - BACKGROUND: Early referral to nephrologists of patients with chronic renal failure (CRF) reduces morbidity and mortality in dialysis. Aim of this work is to evaluate the condition of early and late referral, and whether the two different conditions can affect the treatments. MATERIALS AND METHODS: This is a prospective study with a 12-month follow-up period. During this time, we verified the prevalence of patients with serum creatinine > 1.5 mg/dL (CRF patients) and the condition of early or late referral, defined as referral to nephrologists for > or < 3 times during follow up, respectively. Diagnosis of diabetes mellitus and/or arterial hypertension, and the use of antihypertensive drugs, insulin, hypoproteic diet and erythropoietin was recorded in each patient. RESULTS: CRF (mean serum creatinine value = 2.11+/-1.52 mg/dL) was observed in 190 patients aged 72.05+/-11.62 years. The prevalence of CRF was 4718 pmp. Diabetes and hypertension were diagnosed in 107 subjects (56.3%) and 152 subjects (80%), respectively. Only 74.2% (no. 141) of the patients with CRF was habitually followed by the nephrologist and the frequency was directly correlated to the degree of CRF: 100% of the patients with Creatinine Clearance (Cr Cl) < 25 mL/min, 70% with Cr Cl >25 < 50, and 0% with Cr Cl >50 < 80 mL/min. Early referral was coupled with a wider use of a hypoproteic diet, erythropietin, and the association ACE-I + Angiotensin II receptor antagonists. CONCLUSION: In conclusion, our data show a prevalence of CRF that is at least 5 times greater than that of dialysis patients. The condition of late referral is present in about 30% of the CRF population from the time of the initial phases of renal disease. Referral time affects the modalities of the treatment. PMID- 12746800 TI - [Using Dialysis and Transplantation Registries for regional and small-area epidemiology]. AB - BACKGROUND: Aim of this review on Italian Dialysis and Transplantation Registries (RDTR), mostly of recent implementation, is to assess the use of epidemiological data for planning purposes, in order to promote local developments of regional registries. METHODS; Medline and reports on the website of the Italian Society of Nephrology. RESULTS: We selected two publications with data from the Italian Registry (RIDT) and 56 articles on RDTR. Most papers refer to RDTR for the Italian regions of Piedmont, Lombardy and Lazio. RIDT reports containing 1998 1999 data for all the regions are published on web. Consistent with international figures, epidemiological data show an increasing incidence and prevalence of end stage renal disease patients on renal replacement therapy (RRT), particularly older patients with diabetes and vascular diseases. Important regional variations are observed (RIDT data): 1995-97 dialysis provision (3-25 centres pmp; private: 2%-83%) - 1999 frequency figures: RRT incidence (75-200 pmp); dialysis prevalence (437-842 pmp), transplant prevalence (21-344 pmp) - 1995 mortality (8%-15%). Data on referral modalities and access to RRT seem scarce. Some studies assess hospital use among dialysis patients, while no papers have been found on the decision-making processes in local planning and policy. CONCLUSIONS: RDTR are still under-utilised for developing local data, both for further investigating epidemiological and clinical differences, and for guiding decisions in regions with different resources and cultural background. We believe that linking RDTR data with other regional databases represents an effective strategy. PMID- 12746799 TI - [The importance of intracellular pH in the regulation of cell function]. AB - Cell life is possible only if intracellular pH (pHi) oscillations are kept within a very narrow range. Measurement of pHi is therefore a very important parameter when examining cell and organ functions. Several methods have been used to monitor pHi; these include fluorescent dyes and pH sensitive electrodes. In addition, many instruments have been applied to the detection of pHi in living organs even at the level of single cells: NMR and confocal microscopy are just an example. Transport proteins located on the cell membrane and intracellular vesicles are responsible for maintaining the correct pHi. In renal tubular cells these include the sodium hydrogen exchanger (NHE), the sodium-dependent and independent chloride-bicarbonate exchanger (Cl--HCO3-), the sodium bicarbonate co transport (Na+-HCO3-), the ATP-coupled proton pump (H+-ATPasi) and the ATP dependent proton-potassium pump (H+-K+-ATPasi). All these carriers have been cloned and it is now clear that there are several isoforms with specific properties. The function of these proteins is closely linked to several hormone blood levels, systemic acid-base status, protein diet content as well as other factors. In addition, these carriers are not homogeneously distributed along the nephron and are sensitive to specific stimuli like interstitial osmolality and luminal fluid flow rate. Finally, it has been recently demonstrated that the pHi may be involved in numerous aspects of cell function, such as metabolism, apoptosis, malignancy and it is implicated in the pathogenesis of particular forms of renal stones (Dent's disease). PMID- 12746801 TI - [Morpho-functional study of peritoneum in peritoneal dialysis]. AB - BACKGROUND: Peritoneal dialysis (PD) impairs both structure and function of the peritoneal membrane (PM). Aim of the study is to examine the relationship among dialytic, histological and functional parameters in PD patients. METHODS: Thirty one PD patients were submitted to peritoneal biopsy (PB) during catheter removal for malfunction or after dropping out of the treatment. For each patient PM transport was evaluated by the last peritoneal equilibration test (PET) prior to PB. Each daily glucose load was calculated. PB was performed at a distance of at least 5 cm from the catheter insertion point. The tissues were promptly embedded in formalin and stained for histological and immunohistochemical studies. RESULTS: 1) Patients with mesothelial impairment had longer treatment time. 2) Patients presenting submesothelial sclerosis (SS) and those with impairment of submesothelial basement membrane and subendothelial vascular membrane (SVM) were submitted to larger daily glucose loads. 3) High transporters were exposed to larger daily glucose loads and presented an SS thickness greater than 50 micron more frequently than medium-high transporters. 4) Mesothelial loss was correlated with SS and vascular alterations. 5) SS and vascular injuries were related to each other and not to inflammatory infiltrate. CONCLUSION: Our study suggests that PD length seems to be mainly involved in mesothelial impairment; glucose load appears to damage mainly the submesothelial layer; SS is not a constant event in PD patients and in itself does not seem to be a decisive factor in PD drop out. PMID- 12746802 TI - [Pre-dialysis arteriovenous fistula results in better patency rate]. AB - BACKGROUND: The timing of creation of the first permanent vascular access is crucial to the clinical history of haemodialysis patients. Our strategy is to create vascular access early enough to allow its maturation before the start of the treatment. METHODS: Aim of the study is to evaluate patency of primary A-V fistulas in patients treated between 1985 and 2000 in our dialysis unit. One hundred and thirty A-V fistulas created before haemodialysis treatment (range 10 540 days) and used at its beginning (pre-HD group) are compared with 74 A-V fistulas created and/or used after the start of the haemodialysis treatment (post HD group). RESULTS: Pre-HD group fistulas resulted in higher patency rate than the post-HD group, immediately at the start of the treatment (94.6% vs. 86.5%, p<0.05), at 6 months (89.2% vs. 75.6%, p<0.025), at 12 months (84.5% vs. 64.6%, p< 0.005), at 24 months (77.2% vs. 54.8%, p< 0.005). CONCLUSIONS: A-V fistula is to be preferred in the choice of primary vascular access for chronic haemodialysis patients. It should be created early enough before the beginning of the treatment (when serum creatinine reaches 6 to 7 mg/dL). This planning avoids central venous catheter placement, preserves vessels and the choice of the best surgical option thus resulting in a better fistula survival. PMID- 12746804 TI - [Historical Archives of Italian Nephrology. The history of instrumentation in nephrology. Part II: microscope and haemodialyzer]. AB - Medicine in the technological era acquired many of the characteristics that concurrently marked other fields. So, by adopting procedures based on information obtained with instruments and devices, medicine developed an approach to illness that transformed it into a special form of technology. The collective effect of instrumentation deserves consideration and offers the historian opportunities for interpreting the interaction between physician and his patients in other than scientific and technological terms. The very construction of instruments and devices depends on the Author's ideas assembled with the basic theories of the time. For instance, at the end of the nineteenth century, when medical instruments became essential, the bacterial origin of diseases revolutionised their construction and application. In this context, the invention and use of the microscope became an outstanding feature of the clinical approach by disclosing the cellular universe. The microscope had become crucial in locating some major causes of physical suffering and death in man, and was considered the pre-eminent diagnostic instrument in medicine. In the nephrological field, the microscope drew the physician into a universe of physical changes that were concealed to the naked eye. The microscope made possible the verification of some of Bright's brilliant ideas, something that helped physicians classify glomerulonephritis. Many nephrologists confessed "how few things are established in this subject (nephrology) and how many more difficulties are established, we have learned by experience with the microscope". The modesty of this claim is striking. In nephrology, as in other fields, the admission of ignorance proved to be the beginning of wisdom. This wisdom, based on the admission of ignorance and assembled through the commitment and ingenuity of the pioneers of the dialysis treatment, led to the treatment of end-stage renal disease and the guarantee of success. The technique of haemodialysis has been enriched by the new lexicon, which expresses different ways and ideas on the removal of the solute. PMID- 12746803 TI - [Epstein Barr viral load monitoring in mononuclear lymphocytes and serum of renal transplant recipients using a quantitative PCR protocol]. AB - BACKGROUND: Post-transplant lymphoproliferative disorders (PTLD), ranging from lymphoid hyperplasia to clonal malignancy, are severe complications arising in solid organ transplant patients; their reported incidence ranges from 1 to 20%, according to factors such as type of transplanted organ and age of recipients. A strong correlation between Epstein-Barrvirus (EBV) infection, the grade and type of immunosuppression and the development of PTLD has been recognized. The detection and quantification of EBV-DNA load in peripheral blood have been utilized as prognostic markers for the development of PTLD, showing a correlation between high levels of EBV-DNA in the blood and the development of PTLD. In this study, we monthly monitored EBV viral load in 15 renal transplant recipients for six months. The number of EBV-DNA copies was measured in peripheral blood mononuclear cells (PBMC) and serum samples by a quantitative PCR protocol developed in our laboratory. METHODS: Our EBV-DNA quantification protocol employs a previous screening of samples containing a significant number of viral DNA copies (>=1000 copies/105 PBMC or 100 mL serum) by semi-quantitative PCR followed by a precise quantification of the only significant samples by quantitative competitive (QC)-PCR. RESULTS: Our 15 renal transplant patients neither developed PTLD nor had recurrent acute illnesses or acute graft rejections during the study. The results obtained in the monthly follow up of EB viral load in PBMC samples confirmed its fluctuation in asymptomatic patients reported in the literature. In particular, 5/14 (35.7%) of EBV seropositive patients had an EBV DNA load equal to 1000 EBV copies /105 PBMC, and 1/14 (7.1%) reached 5000 EBV copies /105 PBMC at least once in our study. In the EBV seronegative patient, EBV DNA in PBMC samples was always undetectable (less than 100 DNA copies/105 PBMC). EBV-DNA load in all serum samples was less than threshold value of our quantification protocol (<100 DNA copies/100 mL serum). With regard to the immunosuppressive treatment, it should be noted that 66.7% of the six patients in whom EBV load reached values equal to or higher than 1000 DNA copies/105 PBMC, were on FK506 whereas only 33.3% of them were on CyA. CONCLUSIONS: Since the high positive predictive value of EB viral load in peripheral blood for diagnosis of PTLD reported by several Authors, and the described absence of correlation between the serological evidence of EBV reactivation and EB viral load, EBV viral load measurement in PBMC and serum samples using quantitative PCR techniques is a powerful diagnostic tool to monitor transplanted patients at risk of developing PTLD. PMID- 12746806 TI - [Isolated peripheral arterial ischaemia and medullary neurostimulation: case report]. AB - Isolated peripheral arterial ischaemia (IPAI) is an unusual pathology of dialysis and peritoneal patients which represents the first sign of a complication of uraemia known as calciphylaxis. Recent studies have revealed an increased incidence of this complication. Risk factors are known but there is no consensus on them: elevated CaxP product, female gender, elevated serum parathormone. We present here the case of a 65-year-old man with 21-year history of dialysis, distal isolated ulceration and without any signs of severe vasculopathy. Our clinical diagnosis was calciphylaxis. In this case, the role of early PTX is not clear and the use of steroids is recommended only in non-ulcerating cases. The therapy gives good results but not in all patients. Electrical stimulation of the posterior roots of the spinal cord is an alternative approach to this case. We hypothesised that the electrical action, through cutaneous vasodilatation of afferent dorsal fibres and release of calcitonin gene-releasing protein, determines the release of prostaglandin E sub 2 that may positively affect the proliferation and activity of epidermal fibroblasts. PMID- 12746810 TI - Diamonds from crude oil? PMID- 12746805 TI - [Post-infectious glomerulonephritis]. AB - Post-infectious glomerulonephrites (GNs) include a wide spectrum of nephropathies, with known etiological agent, bacterial, parasitic, viral. Among GNs secondary to bacterial infections, post-streptococcal GN is the most frequent; nevertheless, its incidence in developed countries has decreased during the last 20 years, while some of the characteristics such as types of infection, exposed subjects, clinical and evolutionary patterns have changed. Prognosis has worsened and is correlated with some clinical and histological parameters. The viral infection-related GNs include those associated with HBV, HCV, HIV plus other rarer forms. Membranous GN (MGN), membranoproliferative GN (MPGN) and IgA nephropathy may occur in the course of HBV infection, while different GNs can be detected in relation to HCV, the most frequent being mixed cryoglobulinemic GN, a MPGN with peculiar morphological features. Multiple glomerular involvements are seen from HIV infection, the more characteristic form being the so-called HIV associated nephropathy (HIVAN), a focal segmental glomerulosclerosis with tuft collapse affecting African subjects, which starts with a nephrotic syndrome and rapidly develops into uraemia. Other GNs derive from HIV-related immunecomplexes, some with diffuse proliferative characteristics, or lupus like, with less severe clinical manifestations compared with HIVAN. Among the rare viral infections, we ultimately, mention the association between Parvovirus B19 and "collapsing" focal segmental glomerulosclerosis. PMID- 12746807 TI - ACHEMA & Angewandte. PMID- 12746811 TI - Catalytic synthesis of ammonia-a "never-ending story"? AB - Nitrogen atoms are essential for the function of biological molecules and thus are and important component of fertilizers and medicaments. Bonds to nitrogen also find nonbiological uses in dyes, explosives, and resins. The synthesis of all these materials requires ammonia as an activated nitrogen building block. This situation is true for natural processes and the chemical industry. Knowledge of the various techniques for the preparation of ammonia is thus of fundamental importance for chemistry. The Haber-Bosch synthesis was the first heterogeneous catalytic system employed in the chemical industry and is still in use today. Understanding the mechanism and the translation of the knowledge into technical perfection has become a fundamental criterion for scientific development in catalysis research. PMID- 12746813 TI - A novel 3D brick-wall coordination network based on nodes with square-pyramidal connectivity. PMID- 12746812 TI - Oxazolidinone structure-activity relationships leading to linezolid. AB - The development of bacterial resistance to currently available antibacterial agents is a growing global health problem. Of particular concern are infections caused by multidrug-resistant Gram-positive pathogens which are responsible for significant morbidity and mortality in both the hospital and community settings. A number of solutions to the problem of bacterial resistance are possible. The most common approach is to continue modifying existing classes of antibacterial agents to provide new analogues with improved attributes. Other successful strategies are to combine existing antibacterial agents with other drugs as well as the development of improved diagnostic procedures that may lead to rapid identification of the causative pathogen and permit the use of antibacterial agents with a narrow spectrum of activity. Finally, and most importantly, the discovery of novel classes of antibacterial agents employing new mechanisms of action has considerable promise. Such agents would exhibit a lack of cross resistance with existing antimicrobial drugs. This review describes the work leading to the discovery of linezolid, the first clinically useful oxazolidinone antibacterial agent. PMID- 12746814 TI - Solving larger molecular crystal structures from powder diffraction data by exploiting anisotropic thermal expansion. PMID- 12746815 TI - A highly selective and practical method for enantiopure sulfoxides utilizing activated and functionally differentiated N-sulfonyl-1,2,3-oxathiazolidine-2 oxide derivatives. PMID- 12746816 TI - A colorimetric and ratiometric fluorescent chemosensor with three emission changes: fluoride ion sensing by a triarylborane- porphyrin conjugate. PMID- 12746818 TI - Nanoporous copper silicates with one-dimensional 12-ring channel systems. PMID- 12746817 TI - Isolation and structural proof of the large diamond molecule, cyclohexamantane (C26H30). PMID- 12746819 TI - Oligosaccharide synthesis on a fluorous support. PMID- 12746820 TI - The double N-arylation of primary amines: toward multisubstituted carbazoles with unique optical properties. PMID- 12746821 TI - Enantio- and diastereoselective ir-catalyzed allylic substitutions for asymmetric synthesis of amino acid derivatives. PMID- 12746822 TI - Infrared fingerprint of protonated benzene in the gas phase. PMID- 12746823 TI - Highly stereoselective tandem aza-Michael addition-enolate protonation to form partially modified retropeptide mimetics incorporating a trifluoroalanine surrogate. PMID- 12746824 TI - Vanadium-catalyzed anti-selective additions of allenols to imines. PMID- 12746825 TI - Molecular recognition of polymers by cyclodextrin vesicles. PMID- 12746826 TI - A multicomponent sensing ensemble in solution: differentiation between structurally similar analytes. PMID- 12746827 TI - Synthesis of a C-glycoside analogue of sTn: an HIV- and tumor-associated antigen. PMID- 12746828 TI - [Co(CO)5][(CF3)3BF]: a stable salt of a homoleptic trigonal-bipyramidal metal carbonyl cation. PMID- 12746829 TI - Large electric-field effects on the dipolar aggregation of merocyanine dyes. PMID- 12746830 TI - Drawing small cations into highly charged porous nanocontainers reveals "water" assembly and related interaction problems. PMID- 12746832 TI - Expression study of three secretory proteins (prostatic secretory protein of 94 amino acids, probasin, and seminal vesicle secretion II) in dysplastic and neoplastic rat prostates. AB - BACKGROUND: Prostatic secretory protein of 94 amino acids (PSP94), probasin, and seminal vesicle secretion II (SVSII) are the three major proteins secreted by the lateral lobe of the rat prostate gland. Among these proteins, rodent PSP94 but not probasin and SVSII has a human homologue and it is also a major secretory protein of the human prostate, in addition to prostatic acid phosphatase and prostate-specific antigen. METHODS: In this study, we examined and compared the mRNA expression of these three secretory markers in three rat models of prostate cancer including the sex steroid-induced dysplasia (prostatic intraepithelial neoplasia or PIN) in Noble (Nb) rat model, an androgen-independent Nb rat prostatic tumor (AIT) and Dunning rat prostatic adenocarcinomas (both androgen dependent and -independent) by in situ hybridization (ISH), reverse transcriptase polymerase chain reaction (RT-PCR), and immunohistochemistry. RESULTS: The transcripts for the three markers were highly expressed in the secretory epithelium of normal lateral prostate (LP). Their hybridization signals became reduced in the epithelial cells in the low-grade PINs and significantly weakened or lost in the high-grade PINs induced in the LP. Interestingly, we observed that some dysplastic cells located at the basal compartment of the PIN lesions, and nests of outpouching epithelial cells in the vicinity of PINs, expressed positive hybridization signals of three markers. In the adenocarcinoma, signals of probasin but not PSP94 and SVSII were detected. No hybridization signals were detected in both Dunning and AIT tumors. By RT-PCR, transcripts for these proteins were still detected but significantly reduced in the Dunning tumors, whereas in the AIT tumor, only SVSII transcripts were detected. Immunohistochemistry of PSP94 also showed a reduced staining in the PIN lesions, but no immunoreactivity was seen in the rat prostatic tumors. CONCLUSIONS: The mRNA expression of the three prostatic secretory markers were decreased in the hormone-induced PINs and in two rat prostatic tumors, indicating that the androgen-regulated secretory differentiation was impaired during the development of the premalignant lesion and further reduced in advanced tumors. The abnormal expression pattern of these secretory markers and androgen receptor (AR) in the basal compartment of the PIN lesions suggests that there is a population of cell types with secretory phenotype appearing in the basal cell layer during the early malignant transformation of the prostatic epithelium. PMID- 12746833 TI - Glutathione S-transferase pi is upregulated in the stromal compartment of hormone independent prostate cancer. AB - BACKGROUND: Glutathione S-transferase (GST) pi is a detoxifying enzyme abundant in normal prostate basal cells but only rarely expressed in prostate cancer cells. The current studies are the first to focus on GST pi in the stromal compartment of prostate tumors. METHODS: We employed immunohistochemical, immunofluorescence, and Western blot analysis to measure GST pi expression and subcellular localization in 21 primary and metastatic tumors from patients with hormone independent prostate cancer, as well as seven lymph node metastases and six prostatectomy specimens. RESULTS: GST pi was detectable in stromal cells in 17 of the 21 hormone independent prostate tumors. GST pi tissue distribution in hormone independent tumors coincided with vimentin staining, suggesting that GST pi is expressed by reactive fibroblasts and/or myofibroblasts. CONCLUSIONS: The current results suggest that prostate cancer cells induce an injury response in the stroma during progression to hormone independence, which results in GST pi expression. Stromal GST pi may contribute to chemoresistence of advanced prostate cancer. PMID- 12746834 TI - Possible mechanism of dexamethasone therapy for prostate cancer: suppression of circulating level of interleukin-6. AB - BACKGROUND: Glucocorticoids may have favorable effects on prostate cancer patients showing clinical and/or biochemical failure after androgen ablation. The efficacy and mechanisms of dexamethasone therapy as possible alternative endocrine therapy were investigated. METHODS: Twenty five patients with prostate cancer treated by androgen ablation and showing a steady increase in serum prostate specific antigen (PSA) were treated with low-dose dexamethasone. RESULTS: Of 25 patients, 11 demonstrated 50% or more decline of serum PSA and 9 showed improvement of pain on dexamethasone therapy. Of 8 patients who responded to dexamethasone therapy, 5 had 80% or more decrease in serum interleukin-6 (IL 6). In contrast, none of 8 non-responders showed remarkable IL-6 suppression. Response of PSA was not correlated to the changes in serum dehydroepiandrosterone, dehydroepiandrosterone sulfate, or androstendione. CONCLUSIONS: Significant suppression of serum IL-6, probably through inhibition of androgen-independent activation of androgen receptor, may be one of the mechanisms for the effect of dexamethasone therapy in prostate cancer patients with progressive disease. PMID- 12746835 TI - A novel method of generating prostate cancer metastases from orthotopic implants. AB - BACKGROUND: Spontaneous metastasis following implantation at the orthotopic site is a highly desired feature in prostate cancer (CaP) models, since it would enable studies of mechanisms associated with tumor cell dissemination. METHODS: LuCaP 23.8 and LuCaP 35, hormone-sensitive CaP xenografts that express PSA and the wild-type androgen receptor, were grown orthotopically in SCID mice. When tumor volumes reached approximately 250-500 mg, primary tumors were removed to allow micrometastases to grow. RESULTS: Using this procedure we generated macroscopic metastases (>20 mg) in 71% (LuCaP 23.8) and 100% (LuCaP 35) of animals, respectively. CONCLUSIONS: These models may be used to evaluate new treatment modalities and study mechanisms associated with development of metastases. PMID- 12746836 TI - Insulin-like growth factor binding protein-related protein 1 (IGFBP-rP1/mac 25) is reduced in human prostate cancer and is inversely related to tumor volume and proliferation index in Lucap 23.12 xenografts. AB - BACKGROUND: In the prostate, insulin-like growth factor binding protein-related protein-1 (IGFBP-rP1/mac 25) appears to be decreased in cancer. Likewise, mice injected with prostate cells over-expressing IGFBP-rP1/mac 25 showed decreased tumor formation and tumor volume. METHODS: Immunohistochemistry was used to determine the presence of IGFBP-rP1/mac 25 in human prostate tissue and in LuCaP 23.12 prostate cancer xenografts. RESULTS: Immunoreactivity for IGFBP-rP1/mac 25 was detected in the cytoplasm and nuclei of benign prostate epithelium. Cancer cells also stained but the level of intensity was less than that observed in benign epithelium (P < 0.05). Within the stroma, peripheral nerves, and endothelial vessels reacted with IGFBP-rP1/mac 25 antibodies. IGFBP-rP1/mac 25 was also observed in the nuclei of LuCaP 23.12 cells. LuCaP xenografts with smaller tumor volumes demonstrated more staining, and larger tumor volumes had less staining (P = 0.0033, R = 0.610). There was also a significant inverse correlation between IGFBP-rP1/mac 25 levels in LuCaP 23.12 cells and the proliferative index, measured by percent of cells staining for 5-bromo-2' deoxyuridine (Br-dU) (P = 0.0045, R = 0.594). CONCLUSIONS: IGFBP-rP1/mac 25 is present in normal prostate epithelium and is decreased in human prostate malignancy and higher IGFBP-rP1/mac 25 levels are associated with reduced tumor growth. PMID- 12746837 TI - PRAC2: a new gene expressed in human prostate and prostate cancer. AB - BACKGROUND: The database of human Expressed Sequence Tags was previously used to identify PRAC (Prostate 47:125-131, 2001), a novel gene specifically expressed in human prostate, prostate cancer, rectum, and distal colon. In this report, we have identified PRAC2, another gene with a similar expression pattern that is located adjacent to the original PRAC gene on chromosome 17q21.3. METHODS: Using a computer-based analysis, a cluster of sequence homologous ESTs was identified that is mainly derived from human prostate cDNA libraries. The tissue specificity was examined by multiple tissue RNA dot blots and RT-PCR. The PRAC2 transcript and protein were identified using Northern blot analysis, RACE-PCR, primer extension, and Western blots. RESULTS: PRAC2 encodes a 564 nucleotide RNA found in prostate, rectum, distal colon, and testis. Weak expression was also found in placenta, peripheral blood leukocytes, skin, and in two prostate cancer cell lines: LNCaP and PC-3. The transcript seems to encode a 10.5-kDa nuclear protein. The PRAC2 gene is located on chromosome 17 at position 17q21, between the Hoxb-13 gene and the recently discovered PRAC gene. CONCLUSIONS: Because of the higher expression of PRAC2 in prostate and its proximity to Hoxb-13, PRAC2 may have a function in prostate growth and development. PMID- 12746838 TI - Three immunoassays based on monoclonal antibodies specific for prostate specific antigen (PSA), alpha-1-antichymotrypsin (ACT), and the PSA-ACT complex. AB - BACKGROUND: Prostate specific antigen (PSA) has been widely used as a biomarker for the screening and diagnosis of prostate cancer. PSA in serum predominantly exists as a complex with alpha-1-antichymotrypsin (ACT), and measurement of free PSA and the PSA-ACT complex may improve the utility of the serum PSA assay for differential diagnosis of prostate cancer and non-malignant prostate diseases, such as benign prostatic hyperplasia (BPH). METHODS: Monoclonal antibodies (MAbs) against PSA, ACT, and the PSA-ACT complex were produced by immunizing mice with an incubated mixture of PSA and ACT, and characterized by Western blot analyses and several enzyme-linked immunosorbant assay (ELISA) methods. RESULTS: The MAbs produced in this study are capable of distinguishing the PSA-ACT complex from free PSA and ACT. Four MAbs have been selected and utilized to construct three ELISA systems for the separate measurements of free PSA, the PSA-ACT complex, and total PSA. CONCLUSIONS: The three PSA assay systems developed in this study can specifically measure free PSA, total PSA, and the PSA-ACT complex with equal molar sensitivity. It is expected that these PSA assay systems could be useful in the diagnosis of prostate cancer. PMID- 12746839 TI - ErbB1 and prostate cancer: ErbB1 activity is essential for androgen-induced proliferation and protection from the apoptotic effects of LY294002. AB - BACKGROUND: Androgens play a critical role in proliferation and survival of prostate cancer cells, but the mechanisms leading to these effects are poorly understood. ErbB receptor tyrosine kinases have been implicated in the development of prostate cancer. METHODS: We examined the interaction between the ErbB receptors and androgens using the LNCaP androgen-sensitive prostate tumor model. RESULTS: In the absence of androgens, the cells have low levels of ErbB1 and relatively high levels of ErbB2. Addition of androgens to the medium reversed the ratio; ErbB1 levels rose and ErbB2 levels dropped in response to treatment with the synthetic hormone, R1881. Expression of ErbB activating ligands was found to be constitutive and androgen-independent. The androgen-induced proliferation of LNCaP cells was completely inhibited by the addition of the small molecule ErbB1 tyrosine kinase inhibitors CGP59326 and the bispecific inhibitor (PKI166) for ErbB1 and ErbB2 to the culture medium. Furthermore, in the absence of androgens the relatively low proliferative level was further significantly reduced in the presence of CGP59326. Inhibition of PI3K activity by LY294002 led to induction of apoptosis in androgen-deprived LNCaP cells. Androgen mediated rescue from LY294002-induced apoptosis was inhibited by addition of CGP59326 to the cells. CONCLUSIONS: These results suggest a model whereby androgens promote an increase in the activity of the epidermal growth factor (EGF)-network by increasing ErbB1 levels, and this activity of is essential for androgen-induced proliferation and survival of the prostate cancer LNCaP cell line. PMID- 12746840 TI - Proteomic analysis of human prostasomes. AB - BACKGROUND: Prostasomes are secretory particles in human seminal fluid. Other than a microscopic description of these secretory particles and an incomplete two dimensional gel electrophoresis (2DE) study, little is known about the composition of proteins in prostasomes. METHODS: We employed a direct iterative approach using Gas phase fractionation and microcapillary HPLC-tandem mass spectrometry (microLC-MS/MS) to catalogue the prostasome proteome. RESULTS: We identified 139 proteins that can be divided into the following categories: (1). enzymes (33.8% of total), (2). transport/structural (19.4% of total), (3). GTP proteins (14.4% of total), (4). chaperone proteins (5.8% of total), (5). signal transduction proteins (17.3% of total), and (6). unannotated proteins (9.4% of total). A total of 128 of the 139 proteins have not previously been described as prostasomal. CONCLUSIONS: The proteins identified can be used as reference dataset in future work comparing prostasome proteins between normal and pathological states such as prostate cancer, benign prostatic hyperplasia, prostatitis, and infertility. PMID- 12746841 TI - Anti-proliferative and apoptotic effects of anandamide in human prostatic cancer cell lines: implication of epidermal growth factor receptor down-regulation and ceramide production. AB - BACKGROUND: Anandamide (ANA) is an endogenous lipid which acts as a cannabinoid receptor ligand and with potent anticarcinogenic activity in several cancer cell types. METHODS: The inhibitory effect of ANA on the epidermal growth factor receptor (EGFR) levels expressed on the EGF-stimulated prostatic cancer cells LNCaP, DU145, and PC3 was estimated by ELISA tests. The anti-proliferative and cytotoxic effects of ANA were also evaluated on these human prostatic cancer cell lines by growth tests, flow cytometric analyses, trypan blue dye exclusion assays combined with the Papanicolaou cytological staining method. RESULTS: ANA induced a decrease of EGFR levels on LNCaP, DU145, and PC3 prostatic cancer cells by acting through cannabinoid CB(1) receptor subtype and this leaded to an inhibition of the EGF-stimulated growth of these cells. Moreover, the G(1) arrest of metastatic DU145 and PC3 growth was accompanied by a massive cell death by apoptosis and/or necrosis while LNCaP cells were less sensitive to cytotoxic effects of ANA. The apoptotic/necrotic responses induced by ANA on these prostatic cancer cells were also potentiated by the acidic ceramidase inhibitor, N-oleoylethanolamine and partially inhibited by the specific ceramide synthetase inhibitor, fumonisin B1 indicating that these cytotoxic actions of ANA might be induced via the cellular ceramide production. CONCLUSIONS: The potent anti proliferative and cytotoxic effects of ANA on metastatic prostatic cancer cells might provide basis for the design of new therapeutic agents for effective treatment of recurrent and invasive prostatic cancers. PMID- 12746842 TI - Prostate cancer expression of runt-domain transcription factor Runx2, a key regulator of osteoblast differentiation and function. AB - BACKGROUND: Prostate cancer (CaP) bone metastases express numerous proteins associated with bone cells. Specific transcription factors, including Runx2, regulate the expression of many bone-related factors in osteoblasts. Expression of these transcription factors in CaP may be linked to the ability of CaP bone metastases to influence bone remodeling. METHODS: CaP tissues and cell lines were analyzed for expression of Runx2 mRNA by RT-PCR and in situ hybridization, and protein by immunohistochemistry, Western blotting, and electrophoretic mobility shift assays (EMSA). RESULTS: Runx2 mRNA and protein were detected in CaP tissues and cell lines. A specific Runx2: OSE2 complex could be formed with PC-3 nuclear extracts. CONCLUSIONS: Expression of Runx2 in CaP may be the molecular switch that is associated with expression of various bone-specific factors in CaP. In turn, expression of these factors can influence bone remodeling and possibly play a role in the growth and survival of CaP in bone. PMID- 12746843 TI - Prediction of extraprostatic cancer by prostate specific antigen density, endorectal MRI, and biopsy Gleason score in clinically localized prostate cancer. AB - BACKGROUND: The present study was designed to identify the preoperative parameters, including PSA-based parameters, and endorectal MRI, predictive of pathological stage in males who underwent radical prostatectomy. METHODS: We studied 114 patients who underwent radical retropubic prostatectomy and pelvic lymphadenectomy for clinically localized prostate cancer. Clinical stage was assessed by DRE, pelvic CT scan, endorectal MRI, and bone scan. The correlation between the preoperative parameters, including PSA-based parameters, clinical stage, and histological findings of biopsy specimens, and the pathological stage was analyzed. Logistic regression analysis was performed to identify a significant set of independent predictors for local extent of disease. RESULTS: Seventy-six (66.6%) patients had organ confined cancer and 38 (33.4%) patients had extraprostatic cancer. Of the 38 patients with extraprostatic cancer, four had seminal vesicle involvement, while, none had pelvic lymph node involvement. Biopsy Gleason score, PSA, PSA-alpha1-antichymotrypsin (PSA-ACT), PSA-density (PSAD), PSA-transition zone density, PSA-ACT density, and PSA-ACT transition zone (TZ) density were significantly higher and percent free PSA was lower in the patients with organ confined cancer than those with extraprostatic cancer (P < 0.01). PSAD showed the largest area under the ROC curve (AUC) among those parameters (AUC = 0.732). Sixty-eight (74.7%) of 91 patients with T2 on endorectal MRI had organ confined cancer, while 15 (65.2%) of 23 patients with T3 had extraprostatic cancer (P < 0.01). Multivariate logistic regression analysis indicated that Gleason score (> or =7 vs. < or =6), endorectal MRI findings, and PSAD were significant predictors of extraprostatic cancer (P < 0.01). CONCLUSIONS: The present study demonstrated that preoperative PSAD was the most valuable predictor among PSA-based parameters for extraprostatic disease in patients with clinically localized prostate cancer. The combination of PSAD, endorectal MRI findings, and biopsy Gleason score can provide additional information for selecting appropriate candidates for radical prostatectomy. PMID- 12746844 TI - Luteinizing hormone beta polymorphism and risk of familial and sporadic prostate cancer. AB - BACKGROUND: Circulating testosterone plays an important role in maintenance and growth of prostate cells. Luteinizing hormone (LH), secreted from the anterior pituitary, signals testicular Leydig cells to secrete testosterone. A genetic variant of the LH-beta protein, LH-betaV, exists in up to 40% of Caucasians and is more bioactive than the wild-type protein. We hypothesized that genetically determined variation in LH function might affect susceptibility to prostate cancer via altered testosterone secretion. METHODS: We determined the frequency of the LH-betaV polymorphism (two linked polymorphisms: Trp(8) --> Arg and Ile(15) --> Thr) in familial prostate cancer patients (n = 446), in sporadic prostate cancer patients (n = 388), and in population-based controls without prostate cancer (n = 510) to assess the role of this polymorphism in susceptibility to prostate cancer. RESULTS: A higher frequency of this variant genotype (LH-betaV: Arg(8)/Thr(15)) was observed in familial prostate cancer patients (18.6%) than in controls (13.7%), and after taking into account the correlation of the familial cases and adjusting for age and body mass index (BMI), there was a weak positive association between the variant LH-beta genotype, and risk of familial prostate cancer (OR = 1.29; 95% CI 0.96-1.75). The sporadic case group was also slightly more likely to have a variant genotype (15.2%) compared to the controls (13.7%), and after adjustment for age and BMI, a similar association with this variant was found (OR = 1.33; 95% CI 0.86-02.07). Surgical cases showed a slightly stronger association for the variant LH-beta genotype compared to non-surgical cases, but among the surgical cases there was little variability in risk across nodal status, stage, and tumor grade. CONCLUSIONS: These data are consistent with the hypothesis that the LH-beta variant is a weak risk factor for prostate cancer. PMID- 12746846 TI - Prostate-specific antigen (PSA) protein does not affect growth of prostate cancer cells in vitro or prostate cancer xenografts in vivo. AB - BACKGROUND: Prostate-specific antigen (PSA) is produced in high amounts by normal and malignant prostate cancer cells. PSA is a serine protease with substrates that include semenogelin I and II, insulin-like growth factor binding protein 3, fibronectin, and laminin. PSA, via its enzymatic activity, may play a role in growth, invasion, and metastasis of prostate cancer cells. Recent data also suggest that the PSA protein itself, independent of enzymatic activity, may also function as an endothelial cell-specific inhibitor of angiogenesis. METHODS: Human (PC3, DU145) and rat (AT2, AT6) prostate cancer cell lines were transfected with the full PSA gene encoding preproPSA protein. PSA-producing clones of each cell line were selected and the amount of enzymatically active PSA produced by each cell line determined using a PSA-specific fluorescent peptide substrate. In vitro and in vivo growth characteristics of PSA-producing transfectants were compared to neomycin controls and wild type cells. RESULTS: All selected clones produced and secreted PSA (5-120 ng/ml/10(5) cells). None of the PSA-transfected cell lines produced detectable amounts of enzymatically active PSA. Production of enzymatically inactive PSA by prostate cancer cell lines did not alter growth kinetics in vitro. PSA-producing xenograft doubling times in vivo were similar to neomycin controls and wild type. CONCLUSION: Although recent reports suggest the PSA protein itself may be antiangiogenic, our results demonstrate that production of PSA protein by prostate cancer cells does not significantly alter growth in vitro or in vivo. PMID- 12746845 TI - Evaluation of SRD5A2 sequence variants in susceptibility to hereditary and sporadic prostate cancer. AB - BACKGROUND: The 5 alpha-reductase type II (SRD5A2) catalyzes the conversion of testosterone into the more potent androgen, dihydrotestosterone (DHT), and is thus believed to be the key enzyme for the control of intracellular DHT level in the prostate. Several single nucleotide polymorphisms (SNPs) in the SRD5A2 gene have been found to alter enzymatic activities and were associated with prostate cancer risk or clinical features in several case-control studies. However, the role of SRD5A2 sequence variants in the susceptibility to hereditary prostate cancer (HPC) has not been evaluated to date. METHODS: Three SNPs in the SRD5A2 gene (A49T, V89L, and C682G) and two microsatellite markers near SRD5A2 were genotyped in 159 HPC families to assess their linkage to prostate cancer. In addition, the three SNPs were also genotyped in 245 sporadic cases and 222 unaffected controls to assess their association with hereditary and sporadic prostate cancer. RESULTS: Weak evidence for linkage in the SRD5A2 chromosomal region was observed in the 159 HPC families (HLOD = 0.87, P = 0.04). Stronger evidence for linkage was observed in Caucasian families (HLOD = 1.10, P = 0.02). When stratified by the SNP A49T, no significant evidence for linkage was observed in families with or without the "T" allele. Similarly, family-based association tests failed to observe significant over-transmission of any risk alleles of SNPs A49T, V89L, and C682G to affected offspring. Finally, no significant differences in the distributions of SNPs A49T, V89L, and C682G were found among the HPC probands, sporadic cases, and controls. CONCLUSIONS: Polymorphisms of SRD5A2 are unlikely to significantly increase susceptibility to hereditary or sporadic prostate cancer in the study populations. PMID- 12746847 TI - Anti-androgenic action by red clover-derived dietary isoflavones reduces non malignant prostate enlargement in aromatase knockout (ArKo) mice. AB - BACKGROUND: Red clover (RC)-derived dietary isoflavones have been implicated as potential preventative agents for the development and prevalence of non-malignant prostate diseases. This study investigated whether dietary isoflavones inhibit prostate growth in vivo in the aromatase knock-out (ArKO) mouse that exhibits lifelong elevation of androgens leading to prostate enlargement. METHODS: Adult (11-week-old) wild-type (WT) and ArKO mice were fed on protein matched isoflavones free (IF) and RC (isoflavone rich) diets for 28 days. Individual prostate lobes and testes were weighed and collected for histological analysis and serum androgens were measured. Responses were compared to castration and estrogen administration to ArKO mice to determine the mechanism of action. RESULTS: ArKO mice fed on IF diet exhibited enlarged prostate lobes and elevated serum androgens compared to WT mice. Following 28 days of RC diet, ArKO VP, AP, and SV weights were reduced to WT weights, although testis and body weights remained unaltered. Stereological analysis of VPs revealed a reduction in all components of the tissue, particularly the lumen. The RC diet reduced ArKO serum testosterone and dihydrotestosterone to WT levels. In comparison to castration and estrogen administration, the dietary isoflavones were shown to be anti androgenic rather than weakly estrogenic, mimicking responses observed in the castrated ArKO, rather than estrogen treated ArKOs. CONCLUSIONS: This study demonstrates that RC-derived isoflavones have a significant effect on prostatic growth, and are capable of reducing the enlarged non-malignant prostate phenotype of the adult ArKO mouse, by acting as anti-androgenic agents rather than weak estrogenic substances. PMID- 12746849 TI - Development of a quantitative RT-PCR method to study 5alpha-reductase mRNA isozymes in rat prostate in different androgen status. AB - BACKGROUND: The incidence of prostate cancer (PCa) and benign prostatic hypertrophy (BPH) continues to rise in the Western world. The development and growth of the prostate gland depends on androgen stimulation. Dihydrotestosterone (DHT) is the primary androgen responsible for prostate development and also for the pathogenesis of benign prostatic hyperplasia (BPH). DHT is synthesized in prostate from circulating testosterone (T) through the action of 5alpha-Reductase (5alpha-R) (EC 1.3.99.5), which occurs as two isozymes, type-1 and type-2. Both types are expressed in the prostate: type-2 isozyme is predominantly expressed in prostate and is implicated in BPH and PCa; type-1 isozyme is also increased in some prostatic adenocarcinomas. In recent years, various inhibitors of type-2 isozyme or of both type-1 and type-2 isozyme have been used in prostatic diseases. METHODS: We present the first published measurements of mRNA levels of steroid 5alpha-R isozymes in the ventral prostate of rats of different androgen status. We used a novel method that combines the high specificity of competitive PCR with the sensitivity of laser-induced capillary electrophoresis (LIF-CE). RESULTS: We demonstrated that T and DHT androgens control the expression of both 5alpha-R isozymes in rat prostrate. CONCLUSIONS: This approach could be of great value for the study of prostate diseases in humans and would allow study at the transcriptional level of the effects of drugs that inhibit either or both of these isozymes. PMID- 12746848 TI - Disabled infectious single cycle herpes simplex virus (DISC-HSV) is a candidate vector system for gene delivery/expression of GM-CSF in human prostate cancer therapy. AB - BACKGROUND: DISC-HSV is a replication incompetent herpes simplex virus that is a highly efficient vector for the transduction of genes in vivo and in vitro. We examine the ability of DISC-HSV to infect human prostate cancer cell-lines and xenograft tumor models, and induce expression of reporter and therapeutic cytokine genes. METHODS: Infection was confirmed by cellular staining for the beta-galactosidase reporter gene product, and by EM. Human GM-CSF production following DISC-hGMCSF infection was measured using ELISA. The metabolic activity of infected cells was determined by NADP/NADPH assay. Cell death was estimated by cell-cycle analysis using flow cytometry with propidium iodide staining. RESULTS: Infection of DU145, PC3 and LNCaP cells with DISC-HSV was dose dependent. Cells infected with DISC-hGM-CSF released significant levels of hGM-CSF for 3 days. NADP/NADPH assay suggested that infected cells continued to be metabolically active for 3 days post-infection, which was consistent with flow cytometry findings that cell death did not occur within 7 days of infection. Tumor xenografts injected with DISC-HSV expressed beta-galactosidase, and intracellular viral particles were demonstrated using EM. CONCLUSIONS: We have previously reported the rejection of established tumors following intra-tumoral injection of DISC-GMCSF. This study demonstrates the ability of DISC-HSV to infect prostate cancer and express GMCSF at significant levels. We suggest that prostate cancer is a potential target for therapy using DISC-HSV containing GM-CSF. PMID- 12746850 TI - Isolation, identification and seasonal distribution of airborne and waterborne fungi in Terkos Lake (Istanbul-Turkey). AB - This paper focuses on isolation and identification of airborne and waterborne fungi from different parts of Terkos Lake located in Istanbul (Turkey). The quantitative and qualitative fungal composition of the air and water of the Lake was surveyed monthly for a year (August 2000-July 2001). Water samples were taken at five different stations at Terkos Lake. Airborne fungal spore levels were estimated by exposing a petri dish containing Rose-Bengal streptomycin agar medium to air for 15 minutes. A total of 2372 fungal colonies (1032 from air and 1340 water) was counted on 216 petri plates. We isolated twenty mould species belonging to 9 genera. Scopulariopsis brevicaulis, Penicillium expansum and Cladosporium herbarum were the most abundant species (22.0%, 13.4% and 12.9%, respectively). Cladosporium herbarum and sphaerospermum are very common in air samples (29.7% and 27.0%, respectively). Many of the species isolated are rarely in the atmospheric and water environment such as Aspergillus niger and Cladosporium variabile. Statistical analysis revealed a positive correlation between total CFUs and a number of environmental factors. PMID- 12746851 TI - Chromate tolerance caused by reduced hydroxyl radical production and decreased glutathione reductase activity in Schizosaccharomyces pombe. AB - The stable Cr(VI)-tolerant chr1-66T mutant of Schizosaccharomyces pombe, which carries one simple gene mutation responsible for Cr(VI) tolerance, accumulated and reduced the chromate anion (CrO(4)(2-)) significantly more slowly than did its parental strain 6chr(+). The mutant chr1-66T proved to be sensitive to oxidative stressors such as H(2)O(2), menadione, tert-butyl hydroperoxide and Cd(2+). Both the Cr(VI) tolerance and the oxidative stress sensitivity were attributed to a decreased specific glutathione reductase activity. These effects were also enhanced with a decrease in the specific mitochondrial Mn-SOD activity. PMID- 12746852 TI - An expression system for the functional analysis of pheromone genes in the tetrapolar basidiomycete Schizophyllum commune. AB - The investigation of putative pheromone genes of basidiomycetes has been difficult since the small open reading frames are essentially annotated on the basis of a C-terminal farnesylation signal. In order to identify the functional reading frame, expression of small DNA fragments in the fungus is necessary. The expression system developed in the presented paper allows fusion to the promoter of the tef1 gene encoding the constitutively and highly expressed translation elongation factor EF1alpha. This system has been shown to be functional using an easily selectable gene, ura1. The application to identification of functional pheromone genes has been shown with the newly detected bap2(4) gene. The Bap2(4) pheromone is the first Balpha pheromone gene activating only a single receptor specificity. PMID- 12746853 TI - Transformations of testosterone and related steroids in Absidia glauca culture. AB - In the following study, the course of transformations of testosterone and its derivatives with an additional C(1)-C(2) double bond and/or 17alpha-methyl group or without 19-methyl group in Absidia glauca culture was investigated. The fungi were observed to hydroxylate these compounds and to oxidise the 17beta-hydroxyl group. The products of 6beta, 7alpha, 7beta, 11alpha, 12beta or 15beta hydroxylation were obtained. 19-Nortestosterone was also hydroxylated at the 10beta position.The position and stereochemistry of the introduced hydroxyl group was dependent on the presence of the additional C(1)-C(2) double bond, while hydroxylation at the C-6beta was unaffected. The incubation of testosterone with Absidia glauca gave hydroxyderivatives of androstenedione: 7alpha hydroxyandrostenedione and 6beta,11alpha-dihydroxyandrostenedione. When 1 dehydrotestosterone and 1-dehydro-17alpha-methyltestosterone were used, the products were hydroxylated at 6beta, 7beta or 15beta position while 17alpha methyltestosterone and 19 nortestosterone products were hydroxylated at 6beta, 7alpha, 11alpha or 12beta position. Two metabolites: 6beta,11alpha dihydroxyandrostenedione and 6beta,12beta-dihydroxy-17alpha-methyltestosterone were dihydroxylated. The presence of 17alpha-methyl group in 17alpha methyltestosterone and 1-dehydro-17alpha-methyltestosterone did not influence the position of hydroxylation. PMID- 12746854 TI - Physiological and biochemical response of freshwater cryptomonads (Cryptophyceae) to Fe deficiency. AB - Cryptomonads show strong seasonal distribution in lakes yet little is known of their nutrient requirements. In this study, we examined the role of Fe nutrition in the growth and photosynthetic properties of two freshwater cryptomonads (Cryptomonas sp. UTCC 337 and C. erosa UTCC 446). Cryptomonas sp. appeared more tolerant to Fe deprivation compared with C. erosa. Growth rates calculated for Cryptomonas sp. provided 100 nM and 1 000 nM Fe did not vary (0.55 d(-1)). Only cultures provided 10 nM Fe displayed significantly lower rates of growth (0.26 d( 1)) and lower cellular yields of chlorophyll. In contrast, cultures of C. erosa provided 10 nM Fe failed to grow, whereas cultures provided 100 nM Fe exhibited a reduced rate of growth (30% reduction) and lower yields of cellular chlorophyll (19% reduction) compared to high Fe (1 000 nM) cultures. Photochemical competency, assessed by measuring DCMU-enhanced fluorescence, was high for cells of Cryptomonas sp. regardless of the level of Fe provided (F(v)/F(m) > 0.7). In contrast, photochemical competency was considerably reduced (F(v)/F(m) = 0.46) for C. erosa provided 100 nM Fe. Consistent with this, levels of the Fe containing electron transfer catalyst ferredoxin were reduced by 2.5 times in C. erosa provided 100 nM Fe compared to Fe-replete cells. By comparison, ferredoxin levels varied only slightly in cells of Cryptomonas sp. provided either 100 nM or 1 000 nM Fe. PMID- 12746855 TI - Diagnosis of Mycobacterium tuberculosis in cases of pulmonary infections from Jordanian hospitals. AB - The aim of this study is to use the diagnostic utility of smear technique for the detection of Mycobacterium tuberculosis in clinical specimens obtained from patients suspected of having pulmonary tuberculosis. A total of 305 respiratory specimens (broncoalveolar lavage, sputum and pleural fluid) were collected from 298 patients having pulmonary infections as bronchitis, tuberculosis, pneumonia and other respiratory diseases. Results revealed that 25 specimens collected from 22 patients were positive (8.2%) by acid fast (AF) smear. Data indicated that the specificity and positive predictive value of the conventional smear assay were 100%, however, the sensitivity of the smear examination was 73.5%. Conventional smear technique actually detected M. tuberculosis in respiratory specimens and it could be applied for early and specific diagnosis of tuberculosis in such patients. PMID- 12746856 TI - Protective role of certain chemicals against UV-B-induced damage in the nitrogen fixing cyanobacterium, Nostoc muscorum. AB - The protective effects of L-cysteine, ascorbic acid, reduced glutathione, L tryptophan, and sodium pyruvate against UV-B-induced damages were studied in the nitrogen-fixing cyanobacterium, Nostoc muscorum. When added to the culture suspension during UV-B treatment, these chemicals caused a significant protective effect on survival and growth of the organism. Sodium pyruvate conferred the strongest protection whereas the weakest effect was elicited by tryptophan. A 20 min exposure of a culture suspension to UV-B completely inactivated nitrogenase activity but the inactivation was strongly prevented by exogenous addition of ascorbic acid or reduced glutathione during UV-B exposure, and weakly prevented by pyruvate, cysteine and tryptophan. In vivo nitrate reductase activity was not completely lost even after 80 min of UV-B exposure, and addition of the test chemicals did not confer any significant protection to this enzyme. Whereas (14)CO(2) uptake was drastically inhibited (78% inhibition) by 30 min exposure to UV-B in the absence of any test chemical, about 76% activity remained when the UV B exposure was given to cultures in the presence of ascorbic acid. These results suggest that the damaging effects of UV-B are substantially minimized by certain reducing agents, the protective effect being particularly strong on the O(2) sensitive enzyme, nitrogenase. Presence of these chemicals in their natural habitat or inside the cells of living organisms may partially protect/repair the damaging effects of UV-B radiation. PMID- 12746857 TI - Identification of deoxynivalenol, 3-acetyldeoxynivalenol, and zearalenone in galactose oxidase-producing isolates of Fusarium graminearum. AB - Fusarium graminearum strains are well known for their role as plant pathogens and for their production of mycotoxins, and less known for their secretion of galactose oxidase, a well-studied and useful enzyme. Three galactose oxidase producing isolates of F. graminearum were grown on rice to identify the production of zearalenone and trichothecenes through the use of thin layer chromatography and gas chromatography coupled to mass fragmentation. Detection and identification of deoxynivalenol, 3-acetyldeoxynivalenol, and zearalenone were accomplished. PMID- 12746858 TI - Bacillus thuringiensis associated with faeces of the Kerama-jika, Cervus nippon keramae, a wild deer indigenous to the Ryukyus, Japan. AB - Faeces of the Kerama-jika (Cervus nippon keramae), a wild shika deer indigenous to the Ryukyus, Japan, were examined for the natural occurrence of Bacillus thuringiensis. Of the ten faecal samples tested, seven contained this organism. The frequency of B. thuringiensis was 8.5% among 387 colonies of spore-forming bacteria belonging to the Bacillus cereus/B. thuringiensis group. Of 33 B. thuringiensis isolates recovered, only one isolate, assigned to the serotype H3abc (serovar kurstaki), exhibited dual toxicity against larvae of the silkworm, Bombyx mori, and the mosquito, Aedes aegypti. All of 32 other isolates were serologically untypable or untestable, and were non-toxic to larvae of the two insect species. PMID- 12746859 TI - Quantitative morphologic classification of layer 5 neurons from mouse primary visual cortex. AB - The understanding of any neural circuit requires the identification and characterization of all its components. Morphologic classifications of neurons are, therefore, of central importance to neuroscience. We use a quantitative method to classify neurons from layer 5 of mouse primary visual cortex, based on multidimensional clustering. To reconstruct neurons, we used Golgi impregnations and biocytin injections, as well as DiOlistics, a novel technique of labeling neurons with lipophilic dyes. We performed computerized 3-D reconstructions of 158 layer 5 cells to measure a series of morphologic variables. Principal component analysis and cluster analysis were used for the classification of cell types. Five major classes of cells were found: group 1 includes large pyramidal neurons with apical dendrites that reach layer 1 with an apical tuft; group 2 consists of short pyramidal neurons and large multipolar cells with "polarized" dendritic trees; group 3 is composed of less extensive pyramidal neurons; group 4 includes small cells; and group 5 includes another set of short pyramidal neurons in addition to "atypically oriented" cells. Our sample included a relatively homogeneous group of 27 neurons that project to the superior colliculus, which clustered mainly in group 1, thus supporting the validity of the classification. Cluster analysis of neuronal morphologies provides an objective method to quantitatively define different neuronal phenotypes and may serve as a basis for describing neocortical circuits. PMID- 12746860 TI - Synaptic targets of commissural interneurons in the lumbar spinal cord of neonatal rats. AB - There is strong evidence that commissural interneurons, neurons with axons that extend to the contralateral side of the spinal cord, play an important role in the coordination of left/right alternation during locomotion. In this study we investigated the projections of commissural interneurons to motor neurons and other commissural interneurons on the other side of the spinal cord in neonatal rats. To establish whether there are direct contacts between axons of commissural interneurons and motor neurons, we carried out two series of experiments. In the first experiment we injected biotinylated dextran amine (BDA) into the lateral motor column to retrogradely label commissural interneurons that may have direct projections to motor neurons. Stained neurons were recovered in the ventromedial areas of the contralateral gray matter in substantial numbers. In the second experiment BDA was injected into the ventromedial gray matter on one side of the lumbar spinal cord, whereas motor neurons were simultaneously labeled on the opposite side by applying biocytin onto the ventral roots. BDA injections into the ventromedial gray matter labeled a strong axon bundle that arose from the site of injection, crossed the midline in the ventral commissure, and extensively arborized in the contralateral ventral gray matter. Many of these axons made close appositions with dendrites and somata of motor neurons and also with commissural interneurons retrogradely labeled with BDA. The results suggest that commissural interneurons may establish monosynaptic contacts with motor neurons on the opposite side of the spinal cord. Our findings also indicate that direct reciprocal connections between commissural interneurons on the two sides of the spinal cord may also exist. PMID- 12746861 TI - Characterization of the cDNA encoding a somatostatin variant in the chicken brain: comparison of the distribution of the two somatostatin precursor mRNAs. AB - Although the existence of two somatostatin variants (SS1 and SS2) has now been demonstrated in the brain of mammals, amphibians, and fish, only one isoform of somatostatin (SS1) has been characterized to date in the brain of birds. Here we report cloning of the cDNA encoding a 101-amino-acid protein (PSS2) that encompasses the somatostatin variant [Pro(2)]somatostatin-14 (SS2) at its C terminus. Sequence analysis indicated that chicken PSS2 is more closely related to fish PSS2 than to mammalian cortistatin precursors. Northern blot analysis showed that the chicken PSS1 gene is expressed in the central nervous system (CNS) and in the pancreas, whereas the PSS2 gene is expressed only in the CNS and not in peripheral organs. In situ hybridization histochemistry revealed that, in the chicken brain, PSS1 mRNA is more widely distributed than PSS2 mRNA. In particular, PSS1 mRNA expression was found in the hippocampus, the hyperstriatum, the preoptic area, the ventricular hypothalamic nuclei, the optic tectum, and several nuclei of the mesencephalon and rhombencephalon. In contrast, the distribution of PSS2 mRNA was restricted to a few regions of the brain, including the paraolfactory lobe, the paleostriatum, and some nuclei of the mesencephalon and rhombencephalon. The fact that the PSS1 and PSS2 genes are differently expressed in the brain and in peripheral organs indicates that, in chicken, the two somatostatin variants likely exert distinct functions. In particular, the observation that PSS1 mRNA, but not PSS2 mRNA, occurs in the preoptic area and in the ventral hypothalamic nuclei suggests that, of the two somatostatin isoforms, only SS1 acts as a hypophysiotropic factor. PMID- 12746862 TI - Axonal pathways to the lateral superior olive labeled with biotinylated dextran amine injections in the dorsal cochlear nucleus of rats. AB - The lateral superior olive (LSO) contains cells that are sensitive to intensity differences between the two ears, a feature used by the brain to localize sounds in space. This report describes a source of input to the LSO that complements bushy cell projections from the ventral cochlear nucleus (VCN). Injections of biotinylated dextran amine (BDA) into the dorsal cochlear nucleus (DCN) of the rat label axons and swellings in several brainstem structures, including the ipsilateral LSO. Labeling in the ipsilateral LSO was confined to a thin band that extended throughout the length of the structure such that it resembled an LSO isofrequency lamina. The source of this labeled pathway was not obvious, because DCN neurons do not project to the LSO, and VCN bushy cells were not filled by these injections. Filled neurons in several brainstem structures emerged as possible sources. Three observations suggest that most of the axonal labeling in the LSO derives from a single source. First, the number of labeled VCN planar multipolar cells and the amount of labeling in the LSO were consistent and robust across animals. In contrast, the number of labeled cells in most other structures was small and highly variable. Second, the locations of planar cells and filled axons in the LSO were related topographically to the position of the DCN injection site. Third, labeled terminal arborizations in the LSO arose from collaterals of axons in the trapezoid body (output tract of planar cells). We infer that planar multipolar cells, in addition to bushy cells, are a source of ascending input from the cochlear nucleus to the LSO. PMID- 12746863 TI - Developmental change in expression and subcellular localization of two shaker related potassium channel proteins (Kv1.1 and Kv1.2) in the chick tangential vestibular nucleus. AB - The chick tangential nucleus is a major avian vestibular nucleus whose principal cells participate in two vestibular reflexes. Intracellular recordings have shown that the principal cells acquire their mature firing pattern gradually during development. At embryonic day 16 (E16), most principal cells fire a single spike, whereas shortly after hatching (H) the vast majority fire repetitively on depolarization. The transition in firing pattern was likely due in part to a downregulation of a low-threshold, sustained, dendrotoxin-sensitive (DTX) potassium current, I(DS). Since the DTX-sensitive potassium channel subunits Kv1.1 and Kv1.2 generate sustained currents, in the present study we applied fluorescence immunocytochemistry and confocal microscopy to characterize their developmental expression at E16, H1, and H9. At E16, both Kv1.1 and Kv1.2 staining were confined to the principal cell bodies. Immunolabeling decreased significantly for both proteins at H1, and more so by H9. Double-labeling with a monoclonal antibody against microtubule-associated protein 2 (MAP2) in hatchlings showed that some Kv1.1 remained as clusters within the cell body, at the base of the dendrites, and in the axon initial segment. In hatchlings, Kv1.2 staining decreased in the cell bodies and simultaneously appeared in the neuropil, colocalized with biocytin-labeled primary vestibular fibers and vestibular "spoon" terminals. Also, double-labeling with synaptotagmin showed that Kv1.2 colocalized with many nonvestibular terminals surrounding the principal cell bodies. These results identified developmental decreases in the staining of these two potassium channel protein subunits and changes in their subcellular localization corresponding to the downregulation of I(DS) defined electrophysiologically around hatching. Accordingly, both of these protein subunits could be involved in regulating excitability of the principal cells. PMID- 12746864 TI - Distribution of NTS3 receptor/sortilin mRNA and protein in the rat central nervous system. AB - The neurotensin (NT) receptor, NTS3, originally identified as the intracellular sorting protein sortilin, is a member of a recently discovered family of receptors characterized by a single transmembrane domain. The present study provides the first comprehensive description of the distribution of NTS3/sortilin mRNA and protein in adult rat brain using in situ hybridization and immunocytochemistry. Both NTS3/sortilin mRNA and immunoreactivity displayed a widespread distribution throughout the brain. High levels of NTS3/sortilin expression and immunoreactivity were found in neuronal cell bodies and dendrites of allocortical areas such as the piriform cortex and hippocampus. Regions expressing both high levels of NTS3/sortilin mRNA and protein also included several neocortical areas, the islands of Calleja, medial and lateral septal nuclei, amygdaloid nuclei, thalamic nuclei, the supraoptic nucleus, the substantia nigra, and the Purkinje cell layer of the cerebellar cortex. In the brainstem, all cranial nerve motor nuclei were strongly labeled. NTS3/sortilin mRNA and immunoreactivity were also detected over oligodendrocytes in major fiber tracts. Subcellularly, NTS3/sortilin was predominantly concentrated over intracytoplasmic membrane-bound organelles. Many of the areas exhibiting high levels of NTS3/sortilin (e.g., olfactory cortex, medial septum, and periaqueductal gray) have been documented to contain high concentrations of NT nerve cell bodies and axons, supporting the concept that NTS3/sortilin may play a role in NT sorting and/or signaling. Other areas (e.g., hippocampal CA fields, cerebellar cortex, and cranial nerve motor nuclei), however, are NT-negative, suggesting that NTS3/sortilin also exerts functions unrelated to NT signaling. PMID- 12746865 TI - Autoradiographic analysis of rat brain kinin B1 and B2 receptors: normal distribution and alterations induced by epilepsy. AB - Kindling-induced seizures constitute an experimental model of human temporal lobe epilepsy that is associated with changes in the expression of several inflammatory proteins and/or their receptors in distinct brain regions. In the present study, alterations of kinin receptors in the brain of amygdaloid-kindled rats were assessed by means of in vitro autoradiography, using (125)I-labeled 3-4 hydroxyphenyl-propionyl-desArg(9)-D-Arg degrees -[Hyp(3), Thi(5), D-Tic(7), Oic(8)]-bradykinin (B(1) receptors) and (125)I-labeled 3-4 hydroxyphenyl propionyl-D-Arg degrees -[Hyp(3), Thi(5), D-Tic(7), Oic(8)]-bradykinin (B(2) receptors) as ligands. Results demonstrate that B(2) receptors are widely distributed throughout the brain of control rats. The highest densities were observed in lateral septal nucleus, median preoptic nucleus, dentate gyrus, amygdala, spinal trigeminal nucleus, mediovestibular nucleus, inferior cerebellar peduncles, and in most of cortical regions (0.81-1.4 fmol/mg tissue). In contrast, very low densities of B(1) receptors were detected in all analyzed areas from control rats (0.18-0.26 fmol/mg tissue). When assessed in kindled rats, specific binding sites for B(2) receptors were significantly decreased (41 to 76%) in various brain areas. Conversely, B(1) receptor binding sites were markedly increased in kindled rats, especially in hippocampus (CA2 congruent with CA1 congruent with CA3), Amy and entorhinal, peririnal/piriform, and occipital cortices (152-258%). Data show for the first time that kindling-induced epilepsy results in a significant decline of B(2) receptor binding sites, accompanied by a striking increase of B(1) receptor labeling in the rat brain. An altered balance between B(1) and B(2) receptor populations may play a pivotal role in the onset and/or maintenance of epilepsy. PMID- 12746866 TI - Immunohistochemical distribution of NTS2 neurotensin receptors in the rat central nervous system. AB - In the present study, we localized the levocabastine-sensitive neurotensin receptor (NTS2) protein in adult rat brain by using an N-terminally-directed antibody. NTS2-like immunoreactivity was broadly distributed throughout the rat brain. At the cellular level, the reaction product was exclusively associated with neurons and predominantly, although not exclusively, with their dendritic arbors. No NTS2 signal was observed over astrocytes, as confirmed by dual confocal microscopic immunofluorescence studies using the astrocytic marker S100beta. High densities of NTS2-like immunoreactive nerve cell bodies and/or processes were detected in many regions documented to receive a dense neurotensinergic innervation, such as the olfactory bulb, bed nucleus of the stria terminalis, magnocellular preoptic nucleus, amygdaloid complex, anterodorsal thalamic nucleus, substantia nigra, ventral tegmental area, and several brainstem nuclei. Most conspicuous among the latter were structures implicated in the descending control of nociceptive inputs (e.g., the periaqueductal gray, dorsal raphe, gigantocellular reticular nucleus, pars alpha, lateral paragigantocellular, and raphe magnus), in keeping with the postulated role of NTS2 receptors in the mediation of neurotensin's supraspinal antinociceptive actions. However, the distribution of NTS2-like immunoreactivity largely exceeded that of neurotensin terminal fields, and some of the highest concentrations of the receptor were found in areas devoid of neurotensinergic inputs such as the cerebral cortex, the hippocampus, and the cerebellum, suggesting that neurotensin may not be the exclusive endogenous ligand for this receptor subtype. PMID- 12746867 TI - Contractile cochlear frame in the gecko Teratoscincus scincus. AB - It is generally accepted that the cartilaginous frame of the reptilian cochlea has only a passive supportive function. In this study, a ribbon of contractile tissue was revealed within the cartilaginous frame of the cochlea of the gecko Teratoscincus scincus. It consisted of tightly packed cells and received an extensive blood supply. The cytoplasm of the cells was filled with cytoskeletal filaments 5-7 nm thick as revealed by electron microscopy. Isolated tissue permeabilized with Triton X-100 or glycerol reversibly contracted in the presence of ATP. Noradrenaline caused slow relaxation of the freshly isolated tissue placed in artificial perilymph. We suggest that slow motility of the contractile tissue may adjust passive cochlear mechanics to sounds of high intensities. PMID- 12746869 TI - Ontogeny of ultraviolet-sensitive cones in the retina of rainbow trout (Oncorhynchus mykiss). AB - In order to facilitate emerging models of retinal development, we developed electroretinogram and in situ hybridization protocols to examine the ontogeny of photoreceptors in the retina of a land-locked salmonid, the rainbow trout (Oncorhynchus mykiss). We cloned cDNA fragments corresponding to the rod opsin and each of the four cone opsin gene families, which we utilized to produce riboprobes. We established the specificity of the in situ hybridization protocol by examining subcellular signal localization and through double-labeling experiments. We confirm the assumption that the accessory corner cones in the square mosaic are the ultraviolet wavelength-sensitive (UVS) cone photoreceptor (i.e., they express an SWS1 opsin) and observed UVS cones throughout the retina of small trout. Larger fish have a decrease in sensitivity to short wavelength light stimuli and the distribution of UVS cones in the mature retina is limited to the dorsal-temporal quadrant. These larger fish also possess differentiated UVS cones in the peripheral germinal zone (PGZ), including within areas peripheral to mature retina lacking UVS cones. These data are consistent with the loss of putative UVS cones from the PGZ of a migratory salmonid of another genus, and thus the disappearance of UVS cones appears to be general to the Family Salmonidae, regardless of life history strategy. The generation, differentiation, and subsequent loss of UVS cones in the smolt PGZ is a dramatic example of the supposition that the mechanisms of PGZ development recapitulate the retinal embryogenesis of that species. PMID- 12746871 TI - Expression of metabotropic glutamate receptors in rat meningeal and brain microvasculature and choroid plexus. AB - This study investigated the distribution of metabotropic glutamate receptors (mGluRs) in meningeal and parenchymal microvasculature and in choroid plexus by means of Western blot analysis and immunohistochemistry. Western blot analysis demonstrated mGluR expression in both rat and human leptomeningeal tissues. In the rat, mGluR expression was developmentally regulated, with only mGluR2/3 showing expression at the embryonic day 19 developmental stage. In contrast, mGluR1 alpha, mGluR2/3, mGluR4a, and mGluR7 were expressed in leptomeninges from adult rats. Immunohistochemical analyses showed intense mGluR1 alpha immunoreactivity in the pia mater and blood vessels in the subarachnoid space and in the arachnoid layer of the meninges. mGluR2/3, mGluR4a, mGluR5, and mGluR7 were also expressed in meningeal microvasculature. In addition, the parenchymal microvasculature and choroid plexus were strongly immunoreactive for mGluR1 alpha, mGluR2/3, mGluR4a, mGluR5, and mGluR7. We used antibodies specific for phenotypic markers of microvascular and glial cells to characterize the cell type(s) immunopositive for mGluRs. Comparison of staining with anti-von Willebrand factor antibody and anti-mGluR antibodies revealed that mGluR immunoreactivity was present in cells that surrounded the luminal surface labeled by the endothelial cell marker. In these cells, smooth muscle actin and mGluR immunoreactivity overlapped, suggesting that, in addition to endothelial cells, pericytes within the microvasculature also express mGluRs. Furthermore, expression of mGluR1 alpha was also observed in pure pericyte cultures isolated from bovine retina. These data suggest that glutamate by means of activation of mGluRs may have a broad sphere of physiological influence in the brain which in addition to modulating synaptic transmission may also have a role in determining microvascular function and dysfunction. PMID- 12746870 TI - GFAP knockout mice have increased levels of GDNF that protect striatal neurons from metabolic and excitotoxic insults. AB - In response to injury and degeneration, astrocytes hypertrophy, extend processes, and increase production of glial fibrillary acidic protein (GFAP), an intermediate filament protein located within their cytoplasm. The present study tested the hypothesis that GFAP expression alters the vulnerability of neurons to excitotoxic and metabolic insult induced by 3-nitroproprionic acid (3-NP), an irreversible inhibitor of mitochondrial complex II activity or the excitotoxin quinolinic acid (QA). In this respect, adult GFAP knockout mice (KO) and wild type control mice (WT) received unilateral intrastriatal injections of 3-NP (200 nmol/microl) or QA (100 nmol/microl) and were killed 1, 2, or 4 weeks later. Lesion volume and neuronal counts were quantified using unbiased stereologic principles. For both QA and 3-NP lesions, a significant decrease in lesion volume and an increase in striatal projection neurons were seen in GFAP KO mice compared with WT mice. Enzyme-linked immunoassay analysis revealed increased basal levels of glial cell derived neurotrophic factor (GDNF) relative to WT mice. In contrast, no differences were observed in the expression of ciliary neurotrophic factor or nerve growth factor. These data strongly suggest that the expression of GFAP is implicated with the production of GDNF to a degree that confers neuroprotection after an excitotoxic or metabolic insult. PMID- 12746872 TI - Patterns of expression of neuropeptides in GABAergic nonprincipal neurons in the mouse hippocampus: Quantitative analysis with optical disector. AB - Neuropeptides are widely distributed in the central nervous system and are considered to play important roles in the regulation of neuronal activity. This study shows the patterns of expression of four neuropeptides [neuropeptide Y (NPY), somatostatin (SOM), cholecystokinin (CCK), and vasoactive intestinal polypeptide (VIP)] in gamma-aminobutyric acid (GABA)-ergic neurons of the mouse hippocampus, with particular reference to the areal and dorsoventral difference. First, we estimated the numerical densities (NDs) of GABAergic neurons containing these neuropeptides using the optical disector. The NDs of NPY- and SOM-positive GABAergic neurons were generally higher than those of CCK- and VIP-positive GABAergic neurons. In the whole area of the hippocampus, the ND of NPY-positive GABAergic neurons showed no significant dorsoventral difference (1.90 x 10(3)/mm(3) in the dorsal level, 2.09 x 10(3)/mm(3) in the ventral level), whereas the ND of SOM-positive GABAergic neurons was higher in the ventral level (1.44 x 10(3)/mm(3)) than in the dorsal level (0.80 x 10(3)/mm(3)). The ND of CCK positive GABAergic neurons was also higher in the ventral level (0.57 x 10(3)/mm(3)) than in the dorsal level (0.33 x 10(3)/mm(3)). Similarly, the ND of VIP-positive GABAergic neurons was higher in the ventral level (0.61 x 10(3)/mm(3)) than in the dorsal level (0.43 x 10(3)/mm(3)). Next, we calculated the proportions of GABAergic neurons containing these neuropeptides among the total GABAergic neurons. In the whole area of the hippocampus, NPY-, SOM-, CCK-, and VIP-positive neurons accounted for about 31%, 17%, 7%, and 8% of GABAergic neurons, respectively. The present data establish a baseline for examining potential roles of GABAergic neurons in the hippocampal network activity in mice. PMID- 12746873 TI - Distribution of neuropeptide Y immunoreactivity in the central and peripheral nervous systems of amphioxus (Branchiostoma lanceolatum Pallas). AB - Immunocytochemistry techniques were employed to investigate the distribution of neuropeptide Y-like-immunoreactive (NPY-ir) cells and fibers in the central and peripheral nervous systems of adult amphioxus. NPY-ir neurons of the commissural type were abundant in the brain and present but more scarce in the spinal cord. These neurons gave rise to conspicuous NPY-ir tracts that coursed along the entire length of the nerve cord. Some fibers exhibited conspicuous Herring body like swellings. In the peripheral nervous system, small NPY-ir neurons and a large number of thin, beaded NPY-ir fibers were observed in the atrial region, indicating the involvement of this substance in visceral regulation. A few NPY-ir fibers, possibly afferent to the spinal cord, coursed in the ventral branches of the spinal nerves of this region, whereas no NPY-ir fibers coursed in the preoral or velar nerves or in the dorsal branches of the other spinal nerves. These results indicate that NPY is widely used as a neuroregulator/neurotransmitter in the central and peripheral nervous systems of this primitive chordate. In addition, this study demonstrates the presence of tall, thin NPY-ir cells in the putative adenohypophyseal homologue, the Hatschek's pit organ, which is located in the roof of the preoral cavity (vestibule). PMID- 12746874 TI - Quantitative trait loci modulate ventricular size in the mouse brain. AB - Cerebral ventricular size in humans varies significantly. Abnormal enlargement of the ventricles has been associated with schizophrenia, and hydrocephalus can lead to serious cognitive and motor deficiencies in humans and animals. In this study, we mapped quantitative trait loci (QTLs) modulating cerebroventricular size in mice. We hypothesized that genes underlying hydrocephalus might also modulate normal variation in ventricular size. By using digital images of mouse brain sections and stereological techniques, we estimated the volume of the combined lateral and third ventricles, as well as the volume of the entire brain, in 228 AXB and BXA recombinant inbred mice and their parent strains (A/J and C57BL/6J). Ventricle size, expressed as percentage of brain volume, is a heritable trait (h(2) = 0.32). We detected a major QTL controlling variance in volume on chromosome (Chr) 8 near the markers D8Mit94 and D8Mit189. We also detected a strong epistatic interaction affecting ventricular volume between loci on Chr 4 (near D4Mit237 and D4Mit214) and on Chr 7 (D7Mit178 and D7Mit191). These three QTLs, labeled Vent8a, Vent4b, and Vent7c, are close to genes that have been previously implicated in hydrocephalus. PMID- 12746875 TI - Expression of the genes GAD67 and Distal-less-4 in the forebrain of Xenopus laevis confirms a common pattern in tetrapods. AB - We investigated whether gamma-amino butyric acidergic (GABAergic) cell populations correlate positionally with specific Dlx-expressing histogenetic territories in an anamniote tetrapod, the frog Xenopus laevis. To that end, we cloned a fragment of Xenopus GAD67 gene (xGAD67, expressed in GABAergic neurons) and compared its expression with that of Distal-less-4 gene (xDll-4, ortholog of mouse Dlx2) in the forebrain at late larval and adult stages. In Xenopus, GABAergic neurons were densely concentrated in xDll-4-positive territories, such as the telencephalic subpallium, part of the hypothalamus, and ventral thalamus, where nearly all neurons expressed both genes. In contrast, the pallium of Xenopus generally contained dispersed neurons expressing xGAD67 or xDll-4, which may represent local circuit neurons. As in amniotes, these pallial interneurons may have been produced in the subpallium and migrated tangentially into the pallium during development. In Xenopus, the ventral division of the classic lateral pallium contained extremely few GABAergic cells and showed only low signal of the pallial gene Emx1, suggesting that it may represent the amphibian ventral pallium, homologous to that of amniotes. At caudal forebrain levels, a number of GABAergic neurons was observed in several areas (dorsal thalamus, pretectum), but no correlation to xDll-4 was observed there. The location of GABAergic neurons in the forebrain and their relation to the developmental regulatory genes Dll and Dlx were very similar in Xenopus and in amniotes. The close correlation in the expression of both genes in rostral forebrain regions supported the notion that Dll/Dlx are among the genes involved in the acquisition of the GABAergic phenotype. PMID- 12746876 TI - Abnormal cerebellar histogenesis in PEX2 Zellweger mice reflects multiple neuronal defects induced by peroxisome deficiency. AB - The form and circuitry of the cerebellum develops by a complex process that requires integration of afferent-target interactions between multiple neuronal populations and migratory patterns established by neuron-glial interactions. Analysis of mice lacking the PEX2 peroxisome assembly gene, in which peroxisomal function is disrupted, reveals abnormal cerebellar histogenesis due to the disturbance of multiple cellular processes within neurons. Defects in cerebellar growth and the rostro-caudal foliation pattern reflect a reduced granule neuron population and abnormal Purkinje cell dendrite development. In granule neurons, there is increased apoptotic cell death and delayed movement from the EGL to IGL that reflects cell cycle, maturational and migrational abnormalities. The underlying Purkinje cells have stunted dendrite arbors with abnormal branching patterns, which may reflect altered inductive influences from the delayed granule neuron translocation. A delayed arborization of mutant olivary climbing fibers and their defective translocation from the perisomatic to the dendritic compartment of Purkinje cells results in numerous spines on the soma and proximal dendrites of Purkinje cells. Distal Purkinje cell dendritic spines also display abnormal morphology. These Purkinje cell dendritic abnormalities are seen in association with persistent and enlarged axonal spheroids, further indicating the presence of a degenerative process within the Purkinje cell. This PEX2(-/-) mouse model for the human peroxisomal biogenesis disorder Zellweger syndrome illustrates the complex interplay of abnormal developmental processes in the cerebellum and the importance of peroxisomal function for neuronal migration, proliferation, differentiation, and survival. PMID- 12746877 TI - Interfacial kinetics of titanium- and cobalt-based implant alloys in human serum: metal release and biofilm formation. AB - The biocompatibility of metallic implant surfaces is governed in large part by the interfacial kinetics associated with metal release and protein binding. The kinetics of metal release from, and protein binding to, cobalt- and titanium based implant alloys in human serum were investigated by (1). measuring the temporal release of Cr and Ti into serum from Co-Cr-Mo (ASTM F-75) and Ti implant alloys (Ti-6Al-4V: ASTM F136, and commercially pure Ti, cpTi: ASTM F67), respectively; (2). examining the composition of human serum proteins adsorbed onto the surfaces of Co- and Ti-based implant alloys; and (3). identifying the serum proteins associated with the binding of soluble Cr and Ti degradation products. Analysis of metal dissolution kinetics found that Cr was released from Co-based implant alloy at an order of magnitude higher than Ti was released from Ti-based implant alloys. Serum became saturated with soluble CR and Ti at levels as high as 3250 ng/mL Ti from cpTi; 3750 ng/mL Ti from Ti-6Al-4V; and 35400 ng/mL Cr from Co-Cr-Mo degradation. The observation that human serum binds more released metal from Co-based alloy dissolution was consistent with the observed differences in biofilm composition between the two alloys, where additional serum protein(s) of approximately approximately 140 (kDa) molecular weight were detected on Co-based implant alloy surfaces. However, both Cr and Ti released from Co- and Ti-based alloys exhibited a bimodal binding pattern to both low molecular weight serum protein(s) (<32 kDa), and to higher molecular weight protein(s) in the 180-250 kDa range. Identification of metal alloy-dependent biofilm compositions and dissolution products provides the basis for understanding the bioavailability and bioreactivity of these implant alloys and their degradation products. PMID- 12746878 TI - Functional groups on polystyrene model nanoparticles: influence on protein adsorption. AB - The surface characteristics of intravenously administered particulate drug carriers decisively influence the protein adsorption that is regarded as a key factor for the in vivo fate of the carriers. Latex nanoparticles were synthesized to study the influence of different basic and acidic functional groups on particulate surfaces on the protein adsorption from human serum. The protein mass adsorbed to the particles was assessed by BCA protein assay, the protein adsorption patterns were analyzed by two-dimensional electrophoresis. Considerable differences in the protein adsorption with regard to preferential adsorbed proteins were detectable for the different functional groups. Possible correlations between the surface characteristics and the protein adsorption are shown and discussed. The knowledge concerning the interactions of proteins and nanoparticles can be used for a rational development of particulate drug carriers and can also be useful for an optimized design of medical devices, e.g., hemodialysis membranes or implants. PMID- 12746879 TI - Efficacy of glow discharge gas plasma treatment as a surface modification process for three-dimensional poly (D,L-lactide) scaffolds. AB - Gas plasma surface modification of three-dimensional poly (D,L-lactide) scaffolds fabricated by a novel vibrating particle fabrication technique was demonstrated to enhance cell adhesion, proliferation, and differentiation over 10 days in culture using human embryonic palatal mesenchyme cells. Characterization of corresponding two-dimensional treated surfaces revealed decreased contact angle measurements of 54.2 +/- 0.6 degrees for treated surfaces compared to 72.3 +/- 0.7 degrees for control surfaces (p < 0.05). SEM of treated surfaces revealed increased surface roughness combined with marked pitting and erosion. This may contribute to increased cell adhesion. WST-1 cell proliferation assay measurements as an index of cell numbers revealed a statistically significant increase in proliferation activity on treated surfaces on days 1 and 4 compared with controls. There was a fivefold increase in WST-1 activity for both control and treated groups over 10 days. Confocal laser micrographs revealed increased cell numbers on treated specimens throughout all layers of the scaffold, indicating that the glow discharge process enhanced cell proliferation throughout the entire scaffold architecture. Scanning electron micrographs demonstrated increased cell adhesion for treated specimens at the polymer surface most evident after days 1 and 4 of culture. Alkaline phosphatase (ALP)-specific activity peaked by day 7 for control and treated surfaces, indicating cellular differentiation. There was a trend for increased protein production on the treated specimens compared with controls at the initial time points although the differences were not statistically significant. These results demonstrated that gas plasma surface modification enhances osteoblast-like cell function in a three dimensional scaffold model. PMID- 12746880 TI - Hydroxylapatite and titanium: interfacial reactions. AB - The chemical reactions between hydroxylapatite (HA) and titanium were studied in three different kinds of experiments to increase understanding of how to bond HA to titanium for implant materials. HA powder was bonded to a titanium rod with hot isostatic pressing. Interdiffusion of the HA elements and titanium was found in concentration profiles measured in the electron microprobe. Titanium was vapor deposited on sintered HA discs and heated in air; perovskite (CaTiO(3)) was found on the HA surface with Rutherford backscattering and X-ray diffraction measurements. Powder composites of HA and titanium and TiO(2) were sintered at 1100 degrees C; again, perovskite was a reaction product, as well as beta Ca(3)(PO(4))(2), from decomposition of the HA. These results demonstrate chemical reactions and interdiffusion between HA and TiO(2) during sintering, resulting in chemical bonding between HA and titanium. Thus, cracks and weakness at HA titanium interfaces probably result from mismatch between the coefficients of thermal expansion of these materials. HA composites with other ceramics and different alloys should lead to better thermal matching and better bonding at the interface. PMID- 12746881 TI - A new quantitative method to evaluate the in vitro bioactivity of melt and sol gel-derived silicate glasses. AB - Two melt-derived glasses (45S5 and 60S) and four sol-gel glasses (58S, 68S, 77S, and 91S) have been synthesized. The activation energy for the silicon release was determined, and a very close correlation was observed between this value and published results of the bioactive behavior of the glasses. This relationship can be explained in terms of the influence of chemical composition, textural properties, and structural density on the silanol group formation and silicon dissolution. These measurements provide a quantitative method to evaluate the in vitro bioactivity of SiO(2)-based glasses. Preliminary studies suggest an activation energy gap (Ea) of 0.35-0.5 eV as a boundary between bioactive and nonbioactive glasses. PMID- 12746882 TI - In vitro osteoblast response to anodized titanium and anodized titanium followed by hydrothermal treatment. AB - In this study, Titanium (Ti) surfaces were modified using anodization. The electrolyte used for anodization was a mixture of calcium glycerophosphate and calcium acetate. The anodized surfaces were divided into three groups. Hydrothermal treatments were performed on two of the anodized groups for either 2 or 4 h. In vitro osteoblast response to anodized oxide and the hydrothermal treated oxide after anodization was evaluated in this study. Calcium and phosphorus ions were deposited on the Ti oxide during anodization. Anodized surfaces following a 4-h hydrothermal treatment were observed to promote the growth apatite-like crystals as compared with anodized surfaces after a 2-h hydrothermal treatment. Cellular function and onset of mineralization, as indicated by protein production and osteocalcin production, respectively, also were observed as enhanced on hydrothermal-treated surfaces. It was thus concluded from this study that calcium phosphate and apatite-like crystals could be deposited on Ti surfaces using anodization and a combination of anodization and hydrothermal treatment. It was also concluded that the phenotypic expression of osteoblast was enhanced by the presence of calcium phosphate or apatite-like crystals on anodized or hydrothermally treated Ti surfaces. PMID- 12746883 TI - Effects of multigrooved surfaces on fibroblast behavior. AB - Microgrooves have been investigated as substrates for the control of cell alignment. However, they are relatively too narrow and shallow for controlling the orientation of extracellular matrices (ECM) such as collagen. Multigrooves, a combination of microgrooves and macrogrooves, are expected to be able to control the orientation of both cells and ECM. This study investigated a method for fabricating multigrooves and evaluated fibroblast behavior on these novel surfaces. Multigrooved patterns were fabricated on a gold-alloy metal die, in which 90-degree V-shaped microgrooves with a 2-microm pitch were cut on trapezoidal macrogrooves. The macrogrooves had a 50- microm ridge width, a 50 microm wall width, a 50-microm bottom width, and a 25-microm depth. The grooves were made by an ultraprecision micromachine using a single crystal diamond. This metal die served as a template for making surface replicas from polystyrene. Microgrooved and smooth polystyrene replicas also were prepared as comparative substrates. Mouse fibroblast L929 cells were cultured in each type of replica substrate for 7 to 21 days. After these periods, the cells were fixed with 2.5% glutaraldehyde, treated with conventional methods, and, finally, observed by SEM. Confocal laser scanning microscopy was performed to investigate ECM formation. The multigrooved metal die exhibited the desired sharp configuration without defects. The dimensional values of the multigrooves on the polystyrene replicas were almost the same as the designed values. The fibroblasts on the multigrooved and microgrooved substrates were aligned parallel to the surface grooves after 7 days of incubation. In contrast to the microgrooved and flat surfaces, a dense extracellular matrix was produced along the multigrooves after 21 days of incubation. These results suggest that multigrooves can control the orientation of ECM as well as cells and thus enhance the production of ECM. PMID- 12746884 TI - Production of interferon-beta by fibroblast cells on membranes prepared with RGD containing peptides. AB - The production of interferon-beta by NB1-RGB fibroblast cells cultured on protein and peptide membranes prepared from silk fibroin, motif peptides of silk fibroin [(AG)(n)] containing arginine-glycine-aspartic acid (RGD) peptide, and Pronectin was investigated. The cell density on various protein and peptide membranes was approximately the same, although the production of interferon-beta depended significantly on the membranes where the cells were cultured. The highest production of interferon-beta was observed when the cells were cultured on (AG)(6)RGD(AG)(7) membranes prepared with hexafluoroacetone (HFA) as the casting solvent. On RGD-containing peptide membranes more centrally located in the peptides, the cells produced more interferon-beta when the peptide membranes were prepared with HFA as the casting solvent. However, there was no enhanced production of interferon-beta by cells on (AG)(6)RGD(AG)(7) membranes prepared with 9 mol/L LiBr or 4.5 mol/L LiClO(4) solution as the casting solvent. Therefore, both the chemical composition and the secondary and higher order structure of the peptide membranes are important for enhanced production of interferon-beta. The blocking of integrin beta(1) on the cells by anti-integrin beta(1) antibody prevented the enhanced production of interferon-beta on (AG)(6)RGD(AG)(7) membranes prepared with HFA. We suggest that the cells must bind to the RGD sequence having the appropriate conformation through their integrin beta(1) for enhanced production of interferon-beta. PMID- 12746885 TI - Mannosylerythritol lipids, yeast glycolipid biosurfactants, are potential affinity ligand materials for human immunoglobulin G. AB - Three mannosylerythritol lipids (MEL-A, -B, and -C), yeast glycolipid biosurfactants, were independently attached to poly (2-hydroxyethyl methacrylate) beads (PHEMA), and the three obtained MEL-PHEMA composites were examined for their binding affinity to human immunoglobulin G (HIgG). Of the three composites, the composite bearing MEL-A exhibited the highest binding capacity for HIgG. The binding amount of HIgG increased with increased applied concentration, reaching 106 mg HIgG (per g of composite), with a binding yield of 81%. Interestingly, the protein binding to the composite appeared to follow two different modes (Langmuir type and Freundlich type) depending on the applied concentration. The binding amount of human serum albumin to the composite was much smaller than that of HIgG. The bound human serum albumin, however, had minimal effect on the subsequent binding of HIgG, indicating that the two proteins have different binding sites onto the composite. More significantly, the bound HIgG was efficiently recovered under significantly mild elution conditions: Approximately 90% of the protein was eluted from the composite with phosphate buffer at pH 7. These results indicate that the glycolipid biosurfactant may have great potential as an affinity ligand material for HIgG. PMID- 12746886 TI - The proteoglycan contents of the temporomandibular joint disc influence its dynamic viscoelastic properties. AB - The collagen fibers and proteoglycans in the disc of temporomandibular joint provide resistance to various loadings. Thus far, however, the role of the proteoglycans in determining the viscoelastic properties of the disc has not been investigated. In the present study the hypothesis was tested that the viscoelastic behavior of the disc decreases by the removal of proteoglycans. In 32 bovine discs, dynamic tensile tests with a wide range of frequencies were performed. Before testing, specimens were treated with different concentrations of alpha-amylase to remove proteoglycans. As the frequency increased from 0.1 to 100 Hz, the disc became more viscoelastic. Increasing the concentration of alpha amylase significantly decreased its viscoelasticity. It was concluded that proteoglycans play an important role in determining the viscoelastic properties of the disc and, therefore, give the disc a greater capacity for distributing and reducing stresses. PMID- 12746887 TI - Expression of adhesion molecules and cytokines in vitro by endothelial cells seeded on various polymer surfaces coated with titaniumcarboxonitride. AB - Although endothelial cell (EC) seeding improves the patency of vascular prostheses, the detachment of adherent ECs after the restoration of circulation remains one of the major obstacles. Polymer surfaces for endothelialization can be optimized. In this study, polyethylene terephthalate (PET), polypropylene (PP), polytetrafluoroethylene (PTFE), polyurethane (PUR), and silicone were coated with a titaniumcarboxonitride (Ti(C,N,O)) layer by a plasma-assisted chemical vapor deposition process to verify the effect of titanium onto human saphenous vein ECs. Almost confluent EC monolayers were evaluated for 1) proliferation activity and 2) expression of adhesion molecules using cellular enzyme-linked immunosorbent assay and release of cytokines. The results showed that all titanium-coated polymers and uncoated PET have no toxic effect on human saphenous vein ECs excepting uncoated PTFE, PP, and silicone. Moreover, growing ECs showed an insignificant decrease in cytokine production and an unessential change in basal expression of adhesion molecules. Tumor necrosis factor-alpha induced response depends on polymer surface: for example, intercellular adhesion molecule-1 expression decreased. E-selectin expression was unchanged for culturing ECs on coated PET, PP, and PTFE and reduced for polyurethane and silicone. Vascular cell adhesion molecule-1 expression was unchanged for coated PUR and silicone and reduced for PET, PP, and PTFE. In summary, titanium-coating layers promote adhesion of human ECs on polymer vascular grafts with no proinflammatory reaction of ECs. PMID- 12746888 TI - Calcium-deficient apatite: a first in vivo study concerning bone ingrowth. AB - Biphasic calcium phosphate (BCP) materials are increasingly used to restore bone loss in surgery. Calcium-deficient apatites (CDA), the precursors of BCP, are closer in structure to biological apatites and can be associated with therapeutic agents to form drug-delivery systems. The purpose of this first in vivo study of CDA was to evaluate the osteoconductive properties of two composites, consisting of 40-80 microm granules carried by a cellulose-derived polymer, used to fill critical size bone defects in rabbit femoral ends. Animals were sacrificed 2 or 3 weeks after implantation. Histomorphometric analysis of scanning electron microscopy implant surface files was performed using gray level threshold that distinguish between bone or materials (white) and noncalcified tissue (black). Quantitative results for new bone formation showed no significant differences between the composites or the implantation periods. However, nearly all of the CDA disappeared early while supporting more extensive bone colonization than biphasic calcium phosphates implanted in the same conditions. PMID- 12746889 TI - Macrophage migration inhibitory factor, MIF alleles, and the genetics of inflammatory disorders: incorporating disease outcome into the definition of phenotype. PMID- 12746890 TI - The role of the interleukin-6 family of cytokines in inflammatory arthritis and bone turnover. PMID- 12746891 TI - Endothelin and endothelin receptor antagonists in systemic rheumatic disease. PMID- 12746892 TI - Meta-analysis of four rheumatoid arthritis genome-wide linkage studies: confirmation of a susceptibility locus on chromosome 16. AB - OBJECTIVE: Susceptibility to rheumatoid arthritis (RA) is likely to involve several genes of weak effect, and consequently, individual studies may have insufficient power to detect linkage. Four major RA genome-wide linkage studies have been carried out, but apart from the well-established HLA susceptibility locus, none of the reported significant regions of linkage has been replicated. We applied a genome-search meta-analysis to 4 RA genome searches to assess linkage across studies, using published results. METHODS: For each study, 120 genomic bins of approximately 30 cM were defined and ranked according to the maximum evidence for linkage within each bin. Ranks were summed across studies and each bin was assessed empirically by the magnitude of summed rank, using a permutation test. A high summed rank indicated a region in which evidence for linkage was consistent across several studies. RESULTS: In addition to the HLA locus (P < 0.00002), the strongest evidence for an RA susceptibility locus was found on chromosome 16 (P = 0.004). This locus was not identified as statistically significant in any of the 4 individual RA genome searches. In total, 12 regions achieved a significant (P < 0.05) summed rank, compared with the 6 bins expected by random chance. Four of these regions (on chromosomes 6p, 16cen, 6q, and 12p) reached a significance value of P < 0.01, suggesting that a subset of these regions contains RA susceptibility loci. CONCLUSION: Using a meta analysis approach, we have identified existing and novel putative RA susceptibility loci. These results can provide a basis for further positional and functional candidate-gene studies, and may prove useful in other complex rheumatic diseases. PMID- 12746893 TI - Dynamic gadolinium-enhanced magnetic resonance imaging of the wrist in patients with rheumatoid arthritis can discriminate active from inactive disease. AB - OBJECTIVE: To determine the efficacy of dynamic gadolinium-enhanced magnetic resonance imaging (MRI) of the wrist in the evaluation of disease activity in patients with rheumatoid arthritis (RA). METHODS: Thirty-six patients with RA (with different degrees of disease activity) and 5 healthy controls were studied. MRI was performed with a low-field (0.2T), extremity-dedicated machine. After an intravenous bolus injection of gadolinium-diethylenetriamine pentaacetic acid, 20 consecutive fast spin-echo images of 3 slices of the wrist were obtained every 18 seconds. RESULTS: The curves of synovial membrane enhancement identified the following 2 groups: controls and RA patients in remission, and RA patients with active or intermediately active disease. Both the rate of early enhancement (REE) and relative enhancement (RE) were significantly higher in patients with active RA than in those with inactive RA and controls. The REE and RE were significantly correlated with the number of swollen joints (P < 0.00001 and P = 0.003, respectively), the number of tender joints (P < 0.00001 and P = 0.004, respectively), the Ritchie index (P = 0.0002 for both REE and RE), the Disease Activity Score (P = 0.0004 and P = 0.0008, respectively), the Health Assessment Questionnaire (HAQ) (P = 0.0002 and P = 0.0007, respectively), early morning stiffness (P = 0.001 and P = 0.009, respectively), the C-reactive protein level (P = 0.015 and P = 0.03, respectively), the erythrocyte sedimentation rate (P = 0.03, RE only), and alpha2 globulins (P = 0.036 and P = 0.028, respectively). CONCLUSION: Our data support use of dynamic MRI for discriminating active from inactive RA. Enhancement curves are associated not only with laboratory and clinical indicators of inflammation, but also with the HAQ, a relevant predictor of RA functional outcome. This technique can be repeated frequently and is an excellent candidate for the ideal method for the followup of patients with RA. PMID- 12746894 TI - Role of metacarpophalangeal joint anatomic factors in the distribution of synovitis and bone erosion in early rheumatoid arthritis. AB - OBJECTIVE: To investigate the role of metacarpophalangeal (MCP) joint anatomic and biomechanical factors in the distribution of synovitis and bone erosion in early rheumatoid arthritis (RA). METHODS: Thirty-three patients with early RA with clinically diagnosed MCP joint disease and 28 healthy controls were examined by magnetic resonance imaging of the second to fifth MCP joints of the dominant hand. T1 and T2 fat-suppressed coronal sequences were obtained to assess erosion, and dynamic contrast-enhanced images were acquired to assess synovitis in all of the RA patients and in 8 of the controls. Erosions were defined as bone defects with sharp margins observed using T1-weighted imaging in 2 planes, with a cortical break seen in at least 1 plane. The location of erosions was recorded. The volume of synovitis surrounding each MCP joint (divided into 8 regions) was calculated by summation of voxels derived from the maximal enhancement parameters. The synovial volumes adjacent to MCP joint collateral ligaments were determined by correcting synovial volumes for the positions of asymmetrically placed flexor tendons. RESULTS: In patients with early RA in whom bone erosions were present, there was a propensity for involvement of the radial side of the second (P < 0.0001), third (P = 0.002), and fourth (P = 0.056) MCP joints, but not the fifth. Fifty-two of the 110 erosions (47.3%) occurred adjacent to the radial collateral ligaments of the second, third, and fourth MCP joints. The volume of synovitis was also greater on the radial side of the second (P < 0.0001) and third (P < 0.001) MCP joints. A predilection for synovitis in all of the MCP joints adjacent to the radial collateral ligaments was evident when the positional effects of the flexor tendon were considered. The position of radial collateral ligaments had an effect on erosion formation that was independent of synovitis. A predilection for radial bone damage was also evident in the controls, although lesions were 5-fold less frequent, were generally smaller, and had well-defined margins. CONCLUSION: This study shows that there is a predilection for both synovitis and bone erosion formation on the radial side of the MCP joints in early RA, and that joint inflammation appears to drive the inherent tendency for bone damage on the radial side of joints. These findings have implications regarding the pathogenesis of joint damage in RA. PMID- 12746895 TI - Epstein-Barr virus load in the peripheral blood of patients with rheumatoid arthritis: accurate quantification using real-time polymerase chain reaction. AB - OBJECTIVE: To determine whether patients with rheumatoid arthritis (RA) have elevated Epstein-Barr virus (EBV) load in their peripheral blood mononuclear cells (PBMCs) and whether it is correlated with the HLA-DR genes they express, we developed an accurate EBV DNA quantitative assay using real-time polymerase chain reaction (PCR) with fluorescent probes. METHODS: We studied the EBV DNA load in the PBMCs of 84 patients with RA, 69 normal controls, and 22 patients with rheumatic conditions other than RA. A 214-bp segment from the long internal repeat of EBV was amplified from 500 ng of PBMC DNA (150,000 cells) and quantified by real-time PCR with fluorescent probes. RESULTS: We demonstrated that in patients with RA, the EBV DNA load in PBMCs is increased almost 10-fold compared with that in normal controls. The EBV load is stable over time and is not obviously influenced by disease-modifying antirheumatic drugs or HLA-DR. CONCLUSION: Patients with RA have elevated EBV load in their peripheral blood. PMID- 12746896 TI - Selective elimination of synovial inflammatory macrophages in rheumatoid arthritis by an Fcgamma receptor I-directed immunotoxin. AB - OBJECTIVE: To determine whether monocyte/macrophages from rheumatoid arthritis (RA) patients can be selectively eliminated by a toxin-conjugated antibody CD64 ricin A (CD64-RiA) directed toward the high-affinity receptor for IgG (FcgammaRI), exploiting the capacity of FcgammaRI to efficiently endocytose antibody which it has bound. METHODS: Mononuclear cells from peripheral blood (PB) and synovial fluid (SF) obtained from RA patients were cultured in the presence of CD64-RiA. Cell death of monocyte/macrophages was measured by phenotypic changes (light-scatter patterns and CD14 and FcgammaRI expression) and apoptosis (nuclear DNA fragmentation). We then tested whether CD64-RiA-induced cell death of macrophages affected their capacity to stimulate antigen-induced lymphocyte proliferation and to secrete cytokines. Additionally, the capacity of CD64-RiA to inhibit proinflammatory activity and cartilage degradation by RA synovial tissue explants was evaluated. RESULTS: Inflammatory macrophages from RA SF expressed elevated levels of FcgammaRI and were selectively eliminated by CD64 RiA via apoptotic cell death. Monocyte/macrophages from RA PB, which had lower levels of FcgammaRI expression, were much less affected. Induction of SF macrophage apoptosis was associated with efficient inhibition of antigen-induced lymphocyte proliferation and a reduction in tumor necrosis factor alpha (TNFalpha) release. Consistent with these effects on SF macrophages, CD64-RiA also inhibited TNFalpha production, interleukin-1beta production, and cartilage degrading activity of RA synovial tissue explants. CONCLUSION: Together, these data underscore the crucial role of synovial macrophages in RA joint inflammation and indicate that selective elimination of these cells through FcgammaRI-directed immunotoxins could be a novel approach to the treatment of RA. PMID- 12746897 TI - beta1 integrin-mediated signaling induces intercellular adhesion molecule 1 and Fas on rheumatoid synovial cells and Fas-mediated apoptosis. AB - OBJECTIVE: Rheumatoid arthritis (RA) synovial cells interact with inflammatory cells, as well as extracellular matrices, through integrins. However, the relevance of beta1 integrin to inflammatory processes in RA remains unclear. We examined the role of beta1 integrin-mediated signaling in RA. METHODS: Expression of cell-surface molecules was assessed by FACScan. Engagement of beta1 integrins was performed by crosslinking using a specific monoclonal antibody (mAb) and ligand matrices such as fibronectin or collagen. To determine the involvement of tyrosine kinases in beta1 integrin-mediated signaling, the cells were pretreated with various inhibitors of intracytoplasmic signaling or were transfected with a wild-type focal adhesion kinase (FAK) or a dominant negative truncation of the FAK expression plasmid via cationic liposome-mediated transfection. Apoptosis of synovial cells was detected by double staining with propidium iodide and annexin V. RESULTS: beta1 integrin was highly expressed on RA synovial cells. Engagement of beta1 integrins by crosslinking as well as by ligand matrices markedly up regulated expression of intercellular adhesion molecule 1 (ICAM-1) and Fas. Up regulation of ICAM-1 and Fas induced by beta1 integrin was mediated by the tyrosine kinase signaling pathway, especially involving FAK. Fas-mediated early apoptotic change in the cells was amplified by beta1 crosslinking. CONCLUSION: Our results suggest that interaction of beta1 integrins with extracellular matrix augments expression of ICAM-1 and Fas on RA synovial cells, as well as Fas mediated apoptosis of synovial cells. This might lead to the spontaneous growth arrest through the Fas/Fas ligand pathway observed in RA synovitis. PMID- 12746898 TI - Responses to the proinflammatory cytokines interleukin-1 and tumor necrosis factor alpha in cells derived from rheumatoid synovium and other joint tissues involve nuclear factor kappaB-mediated induction of the Ets transcription factor ESE-1. AB - OBJECTIVE: To investigate the expression of the novel Ets transcription factor ESE-1 in rheumatoid synovium and in cells derived from joint tissues, and to analyze the role of nuclear factor kappaB (NF-kappaB) as one of the central downstream targets in mediating the induction of ESE-1 by proinflammatory cytokines. METHODS: ESE-1 protein expression was analyzed by immunohistochemistry using antibodies in synovial tissues from patients with rheumatoid arthritis (RA) and osteoarthritis (OA). ESE-1 messenger RNA (mRNA) levels were analyzed by reverse transcriptase-polymerase chain reaction or Northern blotting in human chondrocytes, synovial fibroblasts, osteoblasts, and macrophages, before and after exposure to interleukin-1beta (IL-1beta), tumor necrosis factor alpha (TNFalpha), or lipopolysaccharide (LPS) with or without prior infection with an adenovirus encoding the inhibitor of nuclear factor kappaB (IkappaB). The wild type ESE-1 promoter and the ESE-1 promoter mutated in the NF-kappaB site were cloned into a luciferase reporter vector and analyzed in transient transfections. Electrophoretic mobility shift assays (EMSAs) and supershift assays with antibodies against members of the NF-kappaB family were conducted using the NF kappaB site from the ESE-1 promoter as a probe. RESULTS: Immunohistochemical analysis showed specific expression of ESE-1 in cells of the synovial lining layer and in some mononuclear and endothelial cells in RA and OA synovial tissues. ESE-1 mRNA expression could be induced by IL-1beta and TNFalpha in cells such as synovial fibroblasts, chondrocytes, osteoblasts, and monocytes. Transient transfection experiments and EMSAs showed that induction of ESE-1 gene expression by IL-1beta requires activation of NF-kappaB and binding of p50 and p65 family members to the NF-kappaB site in the ESE-1 promoter. Overexpression of IkappaB using an adenoviral vector blocked IL-1beta-induced ESE-1 mRNA expression. Chromatin immunoprecipitation further confirmed that NF-kappaB binds to the ESE-1 promoter in vivo. CONCLUSION: ESE-1 is expressed in synovial tissues in RA and, to a variable extent, in OA, and is specifically induced in synovial fibroblasts, chondrocytes, osteoblasts, and monocyte/macrophages by IL-1beta, TNFalpha, or LPS. This induction relies on the translocation of the NF-kappaB family members p50 and p65 to the nucleus and transactivation of the ESE-1 promoter via a high affinity NF-kappaB binding site. ESE-1 may play a role in mediating some effects of proinflammatory stimuli in cells at sites of inflammation. PMID- 12746899 TI - The pathobiology of focal lesion development in aging human articular cartilage and molecular matrix changes characteristic of osteoarthritis. AB - OBJECTIVE: To determine if early focal lesions seen in aging exhibit molecular changes in the extracellular matrix that are similar to those seen in osteoarthritis (OA) and to examine the interrelationships between matrix degradation and synthesis and how they relate to cartilage turnover. METHODS: Condylar cartilage was obtained postmortem from lesion-free joints and from the lesion (where present as well as) from areas adjacent to and remote from the lesion of 31 knees without signs of joint injury (damage to ligaments or menisci). Cartilage was graded histologically and assayed for type II collagen and proteoglycan aggrecan glycosaminoglycan (GAG) contents and turnover (specifically, type II collagen denaturation and its cleavage by collagenase), type II collagen synthesis (C-propeptide [CPII] content), and aggrecan turnover (846 epitope content). To study the degradation of aggrecan reflected by the release of GAG, we cultured cartilage samples from 10 knees. RESULTS: The more degenerated cartilage from the lesion and adjacent area exhibited significantly more collagen cleavage by collagenase than did cartilage remote from the lesion. Type II collagen denaturation and synthesis were also significantly elevated in the lesion and adjacent cartilage, but neither cleavage nor denaturation correlated with synthesis. Type II collagen content decreased with increasing degeneration, with the lowest levels present in the lesion. Collagen content was indirectly related to denaturation and cleavage adjacent to and remote from the lesion and to denaturation within the lesion. Collagen cleavage and denaturation adjacent to and remote from the lesion were directly interrelated. Cartilage from the lesion contained significantly less GAG than did cartilage adjacent to and remote from the lesion. Aggrecan turnover (846 epitope) was also elevated in both the lesion and adjacent cartilage, whereas GAG release was elevated only in the lesion. GAG and 846 epitope contents were interrelated only at sites remote from the lesion. There was also a direct correlation between collagen and GAG contents in the lesion and in adjacent sites. This correlation was also seen between collagen synthesis (CPII) and the 846 epitope. CONCLUSION: These results demonstrate that lesions seen in aging exhibit molecular changes in matrix turnover similar to those seen in OA articular cartilage at arthroplasty, but not in healthy normal aging cartilage. The direct relationships between type II collagen cleavage and denaturation and the inverse relationship between type II collagen content and cleavage or denaturation implicate collagenase activity and damage to collagen in this loss of collagen during lesion development. The lack of correlation of the increased synthesis with the degradation or content of type II collagen indicates that these aspects of turnover are not coordinated in the pathologic state. However, the direct relationship between collagen and GAG contents in and adjacent to the lesion illustrates the structural interrelationships of collagen and proteoglycan aggrecan molecules. These results suggest that these focal lesions represent the development of early OA and that this involves the progressive damage to articular cartilage surrounding the lesion as part of the process of the development of idiopathic OA. PMID- 12746900 TI - Involvement of CD44 in induction of matrix metalloproteinases by a COOH-terminal heparin-binding fragment of fibronectin in human articular cartilage in culture. AB - OBJECTIVE: To investigate the mechanism of induction of matrix metalloproteinases (MMPs) by a 40-kd COOH-terminal heparin-binding fibronectin fragment (HBFN-f) containing III12-14 and IIICS domains in human articular cartilage in culture. METHODS: Human articular cartilage was removed from macroscopically normal femoral heads and cultured with HBFN-f. MMP secretion into conditioned media was analyzed by immunoblotting (MMPs 1 and 13) and by gelatin zymography (MMPs 2 and 9). Type II collagen cleavage by collagenase was monitored in culture by immunoassay. Involvement of specific peptide-binding domains in HBFN-f and the involvement of CD44 were assessed with synthetic peptides and an anti-CD44 antibody. Immunofluorescence histochemistry was performed using fluorescein isothiocyanate-conjugated anti-CD44 antibody. RESULTS: HBFN-f stimulated production of MMPs 1, 2, 9, and 13 in association with type II collagen cleavage by collagenase in human articular cartilage. Peptide V (WQPPRARI) of HBFN-f, which can bind cell surface heparan sulfate proteoglycan (HSPG), blocked MMP induction by HBFN-f, while the scrambled peptide V (RPQIPWAR) had no effect. Peptide CS-1 of 25 amino acids in IIICS of HBFN-f caused no significant effect. Treatment of cartilage with anti-CD44 antibody or HSPG resulted in significant inhibition of HBFN-f-stimulated MMP production. Preincubation with peptide V blocked binding of the anti-CD44 antibody to chondrocytes in cartilage. CONCLUSION: Interaction of the peptide V sequence in HBFN-f with glycosaminoglycans, such as those in CD44, plays an important role in HBFN-f stimulated MMP production in articular cartilage. Because CD44 is up-regulated in osteoarthritic and rheumatoid arthritic cartilage, the role of the interaction between CD44 and HBFN-f in these pathologies should be of relevance and should be studied further. PMID- 12746901 TI - Insulin-like growth factor 1-induced interleukin-1 receptor II overrides the activity of interleukin-1 and controls the homeostasis of the extracellular matrix of cartilage. AB - OBJECTIVE: We examined the effect of the insulin-like growth factor 1 (IGF-1)/IGF receptor I (IGFRI) autocrine/paracrine anabolic pathway on the extracellular matrix (ECM) of human chondrocytes and the mechanism by which IGF-1 reverses the catabolic effects of interleukin-1 (IL-1). METHODS: Phenotypically stable human articular cartilage cells were obtained from normal cartilage and maintained in culture in alginate beads for 1 week to reach equilibrium of accumulated cell associated matrix (CAM) compounds. Levels of CAM components aggrecan and type II collagen (CII) and levels of intracellular IGF-1, IL-1alpha, and IL-1beta and their respective plasma membrane-bound receptors IGFRI, IL-1 receptor I (IL-1RI), and the decoy receptor IL-1RII were assayed using flow cytometry to investigate the relationship between the autocrine/paracrine pathways and the homeostasis of ECM molecules in the CAM. The effects of IGF-1 on the expression of IGF-1, IL 1alpha, and IL-1beta and their respective receptor systems, the aggrecan core protein, and CII were determined by flow cytometry. RESULTS: Cause-effect relationship experiments showed that IGF-1 up-regulates the levels of IGF-1, IGFRI, aggrecan, and CII in the CAM. No effects on the expression of IL-1alpha and IL-1beta and their signaling receptor IL-1RI were observed. However, IGF-1 was able to reverse IL-1beta-mediated degradation of aggrecan and the repression of the aggrecan synthesis rate. Interestingly, levels of aggrecan and CII in the CAM strongly correlated not only with IGF-1, but also with IL-1RII, which acts as a decoy receptor for IL-1alpha and IL-1beta. This suggests that IGF-1 and IL-1RII may cooperate in regulating ECM homeostasis. Additional experiments demonstrated that IGF-1 up-regulated IL-1RII, thereby overriding the catabolic effects of IL 1. CONCLUSION: These findings reveal a new paradigm by which IGF-1 influences chondrocyte metabolism, by reversing the IL-1-mediated catabolic pathway through up-regulation of its decoy receptor. PMID- 12746902 TI - Proteoglycan degradation after injurious compression of bovine and human articular cartilage in vitro: interaction with exogenous cytokines. AB - OBJECTIVE: Traumatic joint injury leads to an increased risk of osteoarthritis (OA), but the progression to OA is not well understood. We undertook this study to measure aspects of proteoglycan (PG) degradation after in vitro injurious mechanical compression, including up-regulation of enzymatic degradative expression and cytokine-stimulated degradation. METHODS: Articular cartilage tissue explants were obtained from newborn bovine femoropatellar groove and from adult normal human donor knee and ankle tissue. Following injurious compression of the cartilage, matrix metalloproteinase 3 (MMP-3) and MMP-13 messenger RNA (mRNA) expression levels were measured by Northern analysis, and PG loss to the medium after cartilage injury was measured in the presence and absence of added exogenous cytokine (interleukin-1alpha [IL-1alpha] or tumor necrosis factor alpha [TNFalpha]). RESULTS: During the first 24 hours after injury in bovine cartilage, MMP-3 mRNA levels increased 10-fold over the levels in control cartilage (n = 3 experiments), whereas MMP-13 mRNA levels were unchanged. PG loss was significantly increased after injury, but only by 2% of the total PG content and only for the first 3 days following injury. However, compared with injury alone or cytokine treatment alone, treatment of injured tissue with either 1 ng/ml IL 1alpha or 100 ng/ml TNFalpha caused marked increases in PG loss (35% and 54%, respectively, of the total cartilage PG content). These interactions between cytokine treatment and injury were statistically significant. In human knee cartilage, the interaction was also significant for both IL-1alpha and TNFalpha, although the magnitude of increase in PG loss was lower than that in bovine cartilage. In contrast, in human ankle cartilage, there was no significant interaction between injury and IL-1alpha. CONCLUSION: The cytokines IL-1alpha and TNFalpha can cause a synergistic loss of PG from mechanically injured bovine and human cartilage. By attempting to incorporate interactions with other joint tissues that may be sources of cytokines, in vitro models of mechanical cartilage injury may explain aspects of the interactions between mechanical forces and degradative pathways which lead to OA progression. PMID- 12746903 TI - One of two chondrocyte-expressed isoforms of cartilage intermediate-layer protein functions as an insulin-like growth factor 1 antagonist. AB - OBJECTIVE: Aging and osteoarthritic (OA) cartilage commonly demonstrate enhanced expression of the large, transforming growth factor beta (TGFbeta)-inducible glycoprotein cartilage intermediate-layer protein (CILP) as well as enhanced extracellular inorganic pyrophosphate (PPi) that promotes the deposition of calcium pyrophosphate dihydrate crystals. In normal chondrocytes, TGFbeta induces elevated chondrocyte extracellular PPi. Insulin-like growth factor 1 (IGF-1) normally blocks this response and reduces extracellular PPi. However, chondrocyte resistance to IGF-1 is observed in OA and aging. Because CILP was reported to chromatographically fractionate with PPi-generating nucleotide pyrophosphatase phosphodiesterase (NPP) activity, it has been broadly assumed that CILP itself has NPP activity. Our objective was to directly define CILP functions and their relationship to IGF-1 in chondrocytes. METHODS: Using primary cultures of articular chondrocytes from the knee, we defined the function of the previously described CILP (CILP-1) and of a recently described 50.6% identical protein that we designated the CILP-2 isoform. RESULTS: Both CILP isoforms were constitutively expressed by primary cultured articular chondrocytes, but only CILP-1 expression was detectable in cultured knee meniscal cartilage cells. Neither CILP isoform had intrinsic NPP activity. But CILP-1 blocked the ability of IGF-1 to decrease extracellular PPi, an activity specific for the CILP-1 N-terminal domain. The CILP-1 N-terminal domain also suppressed IGF-1-induced (but not TGFbeta-induced) proliferation and sulfated proteoglycan synthesis, and it inhibited ligand induced IGF-1 receptor autophosphorylation. CONCLUSION: Two CILP isoforms are differentially expressed by chondrocytes. Neither CILP isoform exhibits PPi generating NPP activity. But, increased expression of CILP-1, via N-terminal domain-mediated inhibitory effects of CILP-1 on chondrocyte IGF-1 responsiveness, could impair chondrocyte growth and matrix repair and indirectly promote PPi supersaturation in aging and OA cartilage. PMID- 12746904 TI - Plasticity of clonal populations of dedifferentiated adult human articular chondrocytes. AB - OBJECTIVE: To investigate whether adult human articular chondrocytes (AHACs), dedifferentiated by monolayer expansion, can differentiate toward diverse mesenchymal lineages and, if so, whether this ability is regulated by growth factors during monolayer expansion. METHODS: AHACs were expanded as multiclonal or clonal populations in medium without (control) or with factors enhancing cell dedifferentiation (transforming growth factor beta1, fibroblast growth factor 2, and platelet-derived growth factor type BB [TFP]). Cells were then cultured under conditions promoting chondrogenic, osteogenic, or adipogenic differentiation, and the acquired phenotypes were assessed histologically, biochemically, and by real time reverse transcriptase-polymerase chain reaction. RESULTS: Multiclonal populations of both control- and TFP-expanded AHACs differentiated toward the chondrogenic, osteogenic, and adipogenic lineages. Compared with control-expanded AHACs, TFP-expanded cells displayed enhanced chondrogenic differentiation capacity (2.4-fold higher glycosaminoglycan/DNA content and 2,500-fold higher up regulation of type II collagen) and osteogenic differentiation capacity (9.4-fold higher increase in alkaline phosphatase activity and 12.4-fold higher up regulation of bone sialoprotein), but reduced formation of adipocytes (5.2-fold lower oil red O-positive cells/area). Clonal populations of AHACs could be efficiently expanded in TFP, but not in control medium. Most TFP-expanded clones were able to redifferentiate only into chondrocytes (7 of 20) or were unable to differentiate (6 of 20). However, some clones (2 of 20) differentiated toward all of the lineages investigated, thus displaying characteristics of mesenchymal progenitor cells. CONCLUSION: Dedifferentiated AHACs exhibit differentiation plasticity, which is modulated by growth factors used during monolayer expansion and is highly heterogeneous across different clones. Clonal culture of AHACs in the presence of regulatory molecules could lead to the identification of AHAC subpopulations with enhanced cartilage repair capacity. PMID- 12746905 TI - Inflammatory cytokine gene expression in the urinary sediment of patients with lupus nephritis. AB - OBJECTIVE: Lupus nephritis is characterized by intrarenal inflammation and lymphocyte activation. In the present study, the expression of cytokine genes in the urinary sediment of patients with systemic lupus erythematosus (SLE) was examined. METHODS: We studied 3 SLE patient groups (25 with active lupus nephritis [active group], 25 with inactive SLE and previous renal involvement [remission group], 20 with inactive SLE and no history of renal involvement [nonrenal SLE group]) and 2 control groups (10 patients with noninflammatory renal diseases [non-SLE group] and 10 healthy volunteers [healthy group]). Cytokine gene expression in the urinary sediment was studied by real-time quantitative polymerase chain reaction. RESULTS: Expression of interferon-gamma (IFNgamma) in urinary sediment was significantly higher in the active group than in all other groups (P < 0.001 by Kruskal-Wallis test). Among the SLE patient groups, there was a close correlation between IFNgamma expression and the overall SLE Disease Activity Index (SLEDAI) score (Spearman's r = 0.590, P < 0.001) and the SLEDAI renal score (r = 0.642, P < 0.001). Urinary expression of interleukin 2 (IL-2) in the active group was significantly higher than that in the healthy group (P = 0.046) but not in the remission or nonrenal SLE groups. There was no difference in the levels of IL-4 expression among the SLE groups. CONCLUSION: We found a predominance of Th1 cytokine in the urinary sediment of patients with active lupus nephritis. Measurement of cytokine gene expression in urinary sediment may be a useful noninvasive tool for assessing the severity of renal involvement in SLE. PMID- 12746906 TI - Correlation between circulating CD27high plasma cells and disease activity in patients with systemic lupus erythematosus. AB - OBJECTIVE: Disease activity in systemic lupus erythematosus (SLE) is usually assessed with complex disease activity scores comprising a variety of different parameters. In order to determine whether SLE disease activity correlates with abnormal B lymphocyte activity, B cell subsets were analyzed, and their relationship to clinical and humoral measures of disease activity was assessed. METHODS: The distribution of B cell subsets was determined by fluorescence activated cell sorting analysis and assessed in relation to the autoantibody profile, disease activity measured by the SLE Disease Activity Index (SLEDAI) and the European Consensus Lupus Activity Measure scores, disease duration, and therapy. RESULTS: The number and frequency of CD27(high) plasma cells were significantly correlated with the SLE disease activity indices and with the titer of anti-double-stranded DNA (anti-dsDNA) autoantibodies. Circulating B cell subsets were not influenced by age or sex, but appeared to relate to the duration of disease and the therapeutic regimen, with the number and frequency of CD27(high) plasma cells increasing and those of CD27- naive B cells decreasing over time. Patients were divided into those with a SLEDAI score of 0-8 (low disease activity) and those with SLEDAI score >8 (high disease activity). Patients with high disease activity had an increased frequency of both CD19+ B cells and CD27(high) plasma cells. By using a nonparametric data sieving algorithm, we observed that these B cell abnormalities provided predictive values for nonactive and active disease of 78.0% and 78.9%, respectively. The predictive value of the B cell abnormalities (78.9%) was greater than that of the humoral/clinical data pattern (71.4%), including anti-dsDNA antibody levels, circulating immune complexes, increased erythrocyte sedimentation rate, mucocutaneous involvement, and acute renal involvement. CONCLUSION: Flow cytometric monitoring of B cell subsets in the peripheral blood provides new insights into abnormalities of B cell function in SLE and may also be a diagnostically valuable option for monitoring the activity of this autoimmune disease. PMID- 12746907 TI - Increased ubiquitination and reduced expression of LCK in T lymphocytes from patients with systemic lupus erythematosus. AB - OBJECTIVE: To explore regulation of proximal signaling and composition of lipid rafts in T lymphocytes from patients with systemic lupus erythematosus (SLE). METHODS: The expression, phosphorylation, and degradation of lipid raft associated signaling molecules in T lymphocytes from 50 patients with SLE compared with 28 healthy controls and 22 rheumatoid arthritis patients were investigated. Lipid raft and nonraft fractions from T cells were isolated by ultracentrifugation. Proteins in the lipid raft and nonraft fractions were analyzed by Western blotting and probed for phosphotyrosine activity and for LCK, LAT, and CD3 epsilon. Immunoprecipitation experiments were performed to assess protein ubiquitination in T cell lysates. T cell phenotype and levels of intracellular LCK were determined by flow cytometry. RESULTS: LCK, an essential signaling molecule for T cell activation, was significantly reduced in both lipid raft and nonraft fractions of T lymphocytes from patients with active SLE compared with controls, and the reduction was independent of treatment. To identify the likely causes of reduced LCK, we explored the possibility that chronic activation of T lymphocytes underlies LCK degradation. The results revealed an increase in protein ubiquitination, and specifically LCK ubiquitination, in T cells from SLE patients. However, our findings suggest that the increase in ubiquitination is independent of T cell activation. CONCLUSION: LCK is reduced in T cell lipid rafts from patients with SLE. This reduction appears to be independent of activation and may be associated with abnormal ubiquitin-mediated regulation mechanisms. PMID- 12746908 TI - Deposition of nucleosomal antigens (histones and DNA) in the epidermal basement membrane in human lupus nephritis. AB - OBJECTIVE: Antinuclear autoantibodies complexed to nucleosomes can bind to heparan sulfate (HS) in the glomerular basement membrane. This binding is due to the binding of the positively charged histones to the strongly anionic HS. Nucleosomes and histones have been identified in glomerular deposits in human lupus nephritis. We investigated whether nucleosomes are present in the basement membrane of nonlesional skin of lupus patients. METHODS: Skin biopsy samples from patients with systemic lupus erythematosus (SLE) (30 with active lupus nephritis and 15 with inactive disease) and controls (with parapemphigus or diabetes) were stained for IgG, histones, DNA, and nucleosomes. RESULTS: IgG deposits were found in 87% of the patients with lupus nephritis, in 33% of the patients with inactive disease, and in 71% of the parapemphigus patients. Using polyclonal antihistone antibodies, histones were detected in 87% of lupus nephritis patients, but in none of the other SLE patients or the diabetes controls (P < 0.0001). Among the parapemphigus controls, 14% of samples stained positive in one of the polyclonal antihistone stainings (P < 0.0001). Using monoclonal antibodies, histones and DNA were identified in 21% of the lupus nephritis patients. Although none of the other groups showed positive staining for nucleosomes, 7% of the lupus nephritis biopsy samples were positive using antinucleosome monoclonal antibodies. Colocalization of nucleosomal antigens and IgG was confirmed using confocal laser microscopy. CONCLUSION: These findings suggest that nucleosome-mediated binding of autoantibodies to basement membranes may also occur at sites in the body other than in the glomerulus. PMID- 12746909 TI - Correlation of serum anti-DNA topoisomerase I antibody levels with disease severity and activity in systemic sclerosis. AB - OBJECTIVE: To investigate correlations between serum levels of topoisomerase I specific antibody (anti-topo I) and clinical features of systemic sclerosis (SSc), including disease severity (the total skin score [TSS]) and disease activity. METHODS: Using highly sensitive enzyme-linked immunosorbent assays, we measured the levels of anti-topo I antibody, including total IgG, individual IgG subclasses, and IgA, and analyzed their correlations with the TSS in 59 patients with SSc, all of whom had diffuse cutaneous involvement. Serial serum samples were obtained from 11 of these patients. RESULTS: The titers of anti-topo I antibody, including IgG and IgA, were positively correlated with the TSS, a measure of SSc disease severity. In 8 of the 11 patients from whom serial serum samples were obtained, changes in the levels of both IgG and IgA, when detectable, paralleled changes in the TSS. In 3 patients, an increasing anti-topo I IgG level preceded an increase in the TSS. The level of each IgG subclass also correlated with and tended to parallel the TSS. The patients with very active disease had higher mean IgG (P < 0.001) and IgA (P < 0.05) titers than did those with inactive disease. CONCLUSION: Serum levels of anti-topo I antibody correlate positively with disease severity and disease activity in SSc. PMID- 12746910 TI - Anatomic structures involved in early- and late-stage sacroiliitis in spondylarthritis: a detailed analysis by contrast-enhanced magnetic resonance imaging. AB - OBJECTIVE: To localize inflammatory and chronic changes to defined areas in the sacroiliac joints in patients with early-stage compared with late-stage spondylarthritis (SpA), using magnetic resonance imaging (MRI). METHODS: Using MRI, 93 patients with SpA and inflammatory back pain who had radiographs of the sacroiliac joints were examined, comprising 31 patients with ankylosing spondylitis (AS) and 62 with other SpA subsets, including 48 with undifferentiated SpA (uSpA). MRI was performed using T1-weighted, T2*-weighted, STIR, and dynamic contrast-enhanced (gadolinium diethylenetriaminepentaacetic acid) sequences. Two readers retrospectively analyzed the images by differentiating 9 areas of the sacroiliac joints: the ventral and caudal joint capsule, cavum, subchondral bone, bone marrow, ligament entheses, and ligaments; the sacral and iliac sides were tabulated separately. RESULTS: By MRI, sacroiliitis was more often bilateral in AS (84%) than in uSpA (48%) (P = 0.01). Inflammatory changes were found in a mean +/- SD 4.7 +/- 2.9 regions/joint, with involvement of 4.5 +/- 3.2 regions in early disease versus 5.2 +/- 2.3 regions in late disease (P not significant [NS]). Involvement of the iliac side of the sacroiliac joints was found to be more frequent than the sacral side in early disease (58% versus 48%; P < 0.01) as compared with that in late disease (58% versus 63%; P NS). The dorsocaudal parts of the synovial joint and the bone marrow were the most frequently inflamed structures in early disease (P < 0.001 for ventral versus dorsal joint capsule). In contrast, involvement of the entheses was more common in advanced disease (early 43% versus late 86%; P < 0.001). Similarly, the ligaments were more frequently involved in the late stages (early 26% versus late 40%; P = 0.06). Both patterns of bone marrow inflammation (focal and diffuse) were observed in equal frequencies in early and late disease (17% and 42% versus 26% and 43%, respectively; P NS). HLA-B27-positive patients (n = 80) had more entheseal involvement than did HLA-B27-negative patients (n = 13) (60% versus 39%; P = 0.05). HLA-B27-negative patients had a shorter disease duration (2.2 years versus 4.4 years; P = 0.05) and were more often female (62%; P = 0.02). When all pathologic changes were assessed, the STIR sequence (performed in 62 patients) was less sensitive than the contrast-enhanced sequences in that it was not able to show all relevant changes in 27% of these patients (n = 17), failing to reveal inflammation of the cavum in 15 patients and of the bone marrow and joint capsule in 1 patient each. CONCLUSION: As visualized by MRI, sacroiliitis in SpA is characterized by involvement of different joint structures. Whereas the iliac and the sacral side of the sacroiliac joints are almost equally affected, the dorsocaudal synovial part of the joint is involved significantly more often than the ventral part, especially in early disease. Sacroiliac enthesitis is not a special feature of early sacroiliac inflammation. PMID- 12746911 TI - Two distinctive HLA haplotypes harbor the B27 alleles negatively or positively associated with ankylosing spondylitis in Sardinia: implications for disease pathogenesis. AB - OBJECTIVE: To compare haplotype distribution in HLA-B27-positive patients with ankylosing spondylitis (AS) and healthy control subjects possessing either AS associated HLA-B27 alleles or the non-AS-associated HLA-B*2709 allele. METHODS: DNA samples from 47 HLA-B27-positive patients with AS and 76 HLA-B27-positive healthy controls (19 positive and 57 negative for B*2709) living in different areas of Sardinia were collected and typed for HLA class I and class II alleles. The third exon of the B27 gene was analyzed for the presence of Asp(116) or His(116), which differentiates B*2709 from the other two B27 subtypes (B*2705 and B*2702) that are mostly found in Sardinia. The parents of 6 subjects positive for B*2709 were also typed for HLA class I and class II alleles. Statistical analysis was performed by Fisher's exact test. RESULTS: In Sardinia, the B27 alleles conferring susceptibility to AS appear to be more frequently carried by a haplotype (A2;B27;Cw2;DR16) that reaches its highest frequency in patients with AS (A2 80.8%, B27 100%, Cw2 83%, and DR16 74.5%). Conversely, the non-AS associated B*2709 allele is more frequently found together with other HLA alleles whose frequencies are inversely correlated with the disease (A32 or A30, Cw1, and DR12). Familial analysis of 6 subjects positive for HLA-B*2709 confirmed the existence of a "Sardinian" haplotype that is not associated with AS (A32;B*2709;Cw1;DR12). CONCLUSION: In Sardinia, 2 distinct haplotypes harbor the non-AS-associated HLA-B*2709 allele or the AS-associated B27 alleles. Our findings are compatible with the hypothesis that other genes within the HLA region besides HLA-B27 may play some role in conferring susceptibility to AS. PMID- 12746912 TI - Daily pain and symptoms in children with polyarticular arthritis. AB - OBJECTIVE: To analyze patterns of daily pain, stiffness, and fatigue related to juvenile arthritis; to examine the relationships of demographics, disease severity, and psychological adjustment to daily disease symptoms; and to examine daily disease symptoms as predictors of reduced participation in school and social activity. METHODS: For a 2-month period, 41 children with polyarticular juvenile arthritis completed daily diaries that included measures of symptoms and function. Children also underwent an initial evaluation and 4 followup evaluations that included a joint count, laboratory testing, and completion of questionnaires assessing physical and psychosocial functioning. RESULTS: Children reported having pain an average of 73% of days, with the majority of children (76%) reporting pain on >60% of all days. On average, children described the intensity of their daily pain as being in the mild to moderate range; however, a significant subgroup (31%) reported pain in the severe range. Higher physician global assessment ratings, increased functional disability, and increased anxiety were significantly associated with increased daily pain and other daily symptoms. Multilevel random-effects analyses indicated that increased daily symptoms of pain, stiffness, and fatigue were significant predictors of reduced participation in school and social activities. CONCLUSION: Physicians should consider treating pain more aggressively in children with arthritis, in order to preserve function in school and social domains, as well as physical function. Moreover, optimal pain management in children with arthritis should include therapeutic regimens addressing anxiety as well as standard pharmacologic interventions. PMID- 12746913 TI - Functional and prognostic relevance of the -173 polymorphism of the macrophage migration inhibitory factor gene in systemic-onset juvenile idiopathic arthritis. AB - OBJECTIVE: To address the functional and prognostic relevance of the -173 single nucleotide G-to-C polymorphism of the macrophage migration inhibitory factor (MIF) gene in patients with systemic-onset juvenile idiopathic arthritis (systemic-onset JIA) by evaluating its association with serum and synovial fluid levels of MIF, with glucocorticoid requirement, and with the outcome of the disease. METHODS: A total of 136 patients with systemic-onset JIA were studied, including 98 patients from the British Paediatric Rheumatology Study Group's National Repository for JIA and 38 patients who were followed up at the IRCCS Policlinico San Matteo (Pavia, Italy) and the IRCCS G. Gaslini (Genoa, Italy). The MIF-173 polymorphism was genotyped using SnaPshot ddNTP primer extension and capillary electrophoresis. MIF levels were measured by enzyme-linked immunosorbent assay. The evaluation of the association of the MIF-173 polymorphism with outcome was performed only in Italian patients who were followed up for >5 years, by analyzing retrospectively 1) the number of joints with active arthritis and the number of joints with limited range of motion; 2) the score, at the last visit, on the Italian version of the Childhood Health Assessment Questionnaire (C-HAQ); and 3) data concerning the treatment regimens during the disease course. RESULTS: Systemic-onset JIA patients carrying a MIF 173*C allele had serum and synovial fluid levels of MIF significantly higher than those in patients with the GG genotype. The duration of glucocorticoid treatment on a daily regimen was significantly longer in patients carrying a MIF-173*C allele than in MIF-173 GG homozygous patients. Moreover, the duration of clinical response to intraarticular injection of triamcinolone hexacetonide was significantly shorter in patients carrying a MIF-173*C allele. At the last visit, the numbers of joints with active arthritis, the C-HAQ scores, and the numbers of joints with limited range of motion were significantly higher in patients carrying the MIF-173*C allele. CONCLUSION: Our study shows the functional relevance of the MIF-173 polymorphism and suggests that the MIF-173*C allele is a predictor of poor outcome in systemic-onset JIA. PMID- 12746914 TI - Cytokine gene polymorphisms: association with psoriatic arthritis susceptibility and severity. AB - OBJECTIVE: To determine whether functional cytokine gene polymorphisms influence disease susceptibility and phenotype in patients with psoriatic arthritis (PsA). METHODS: DNA was obtained from 147 PsA patients and 389 controls. Seven functional proinflammatory (interleukin-1beta [IL-1beta] +3953, IL-6 -174, tumor necrosis factor alpha [TNFalpha] -308, TNFbeta +252) and antiinflammatory (IL-10 1082, IL-10 -592, IL-1 receptor antagonist [intron 2, 86 bp, variable-number tandem repeat]) gene polymorphisms were detected by polymerase chain reaction and restriction fragment length polymorphism assays. RESULTS: No significant difference in genotype frequencies was observed between the control and the PsA patient populations, and no association with Steinbrocker functional class, disease classification (polyarticular or oligoarticular), presence of spinal involvement, or age at PsA onset was observed. The presence of joint erosions was significantly associated with the TNFalpha -308 and TNFbeta +252 polymorphisms (P < 0.0001 and P = 0.0017, respectively). Frequencies of the TNFalpha -308 and TNFbeta +252 genotypes were also significantly different (P = 0.0078 and P = 0.0486, respectively) in a group of progressors (patients with early PsA in whom the number of joint erosions in the hands and feet increased over a median interval of 24 months) compared with a group of nonprogressors. Age at psoriasis onset was significantly associated with the TNFbeta +252 and TNFalpha -308 polymorphisms (P = 0.0003 and P = 0.0081, respectively). The TNFB2B2 and TNFalpha -308 AA genotypes were associated with the earliest mean ages at psoriasis onset. CONCLUSION: The TNFalpha -308 and TNFbeta +252 polymorphisms were significantly associated with age at psoriasis onset, presence of joint erosions in PsA, and progression of joint erosions in early PsA. TNF gene polymorphisms may be useful prognostic markers in PsA, and these results support the rationale for using anti TNF treatment in patients with severe, progressive PsA. PMID- 12746915 TI - Intravertebral clefts in osteoporotic vertebral compression fractures. AB - OBJECTIVE: To describe the characteristics, radiographic appearance, and significance of intravertebral clefts in vertebral compression fractures (VCFs) of patients with osteoporosis presenting for vertebroplasty. METHODS: This was a prospective radiographic study of 50 consecutive patients with 82 VCFs who underwent vertebroplasty at a tertiary referral center. Patients underwent imaging preoperatively with standing lateral and supine cross-table lateral radiographs and magnetic resonance imaging (MRI). Standing lateral radiographs were also obtained postoperatively. Clefts were defined at the time of vertebroplasty as confluent reservoirs for polymethylmethacrylate (PMMA). Postoperatively, all images were reexamined for the presence and characterization of intravertebral clefts. RESULTS: Twenty-four of 50 patients (48%) had clefted VCFs, and 30 of 82 VCFs (37%) contained clefts. Clefted VCFs were severe, dynamically mobile, and occurred primarily in the thoracolumbar junction. Clefts were detectable by standing lateral radiography in 14%, by supine cross-table radiography in 64%, and by MRI in 96% of fractured vertebrae. Clefts occurred primarily in the anterosuperior portion of the vertebral body, and cleft margins appeared increasingly sclerotic in persistently mobile VCFs. PMMA fill patterns of clefted and unclefted VCFs were distinct. CONCLUSION: Intravertebral clefts occur frequently in osteoporotic VCFs of patients who present for vertebroplasty. The radiographic characteristics of clefts evolve over time and can be indistinguishable from Kummell's disease in severe, persistently mobile, clefted osteoporotic VCFs. PMID- 12746916 TI - Neurophysiologic evidence for a central sensitization in patients with fibromyalgia. AB - OBJECTIVE: To determine whether abnormalities of peripheral and central nociceptive sensory input processing exist outside areas of spontaneous pain in patients with fibromyalgia (FM) as compared with controls, by using quantitative sensory testing (QST) and a neurophysiologic paradigm independent from subjective reports. METHODS: A total of 164 outpatients with FM who were attending a self management program were invited to participate in the study. Data for 85 patients were available and were compared with those for 40 non-FM controls matched for age and sex. QST was performed using thermal, mechanical, and electrical stimuli at locations of nonspontaneous pain. Pain assessment was 2-fold and included use of subjective scales and the spinal nociceptive flexion reflex (NFR), a specific physiologic correlate for the objective evaluation of central nociceptive pathways. Questionnaires regarding quality of life and the impact of FM were available. RESULTS: Participants were mainly middle-aged women, with a mean disease duration of 8 years. Between-group differences were significant for neurophysiologic, clinical, and quality of life measures. In patients with FM, peripheral QST showed significantly altered cold and heat pain thresholds, and tolerance to cold pain was radically reduced. The median NFR threshold in patients with FM (22.7 mA [range 17.5-31.7]) was significantly decreased compared with that in controls (33 mA [range 28.1-41]). A cutoff value of <27.6 mA for NFR provided sensitivity of 73% and specificity of 80% for detecting central allodynia in the setting of FM. CONCLUSION: Our results strongly, although indirectly, point to a state of central hyperexcitability of the nociceptive system in patients with FM. The NFR can be used to assess central allodynia in FM. It may also help discriminate patients who may benefit from use of centrally acting analgesics. PMID- 12746917 TI - Amyloid-enhancing factor mediates amyloid formation on fibroblasts via a nidus/template mechanism. AB - OBJECTIVE: To determine the mechanism by which amyloid-enhancing factor (AEF) promotes amyloid deposition, and to test whether AEF seeds deposition of serum amyloid A (SAA) and facilitates conversion to beta-sheet structure. METHODS: Fibroblasts were cultured with mouse recombinant SAA1.1 and AEF, SAA1.1, or AEF. AEF was prepared as a glycerol extract of spleen from amyloidotic mice. Amyloid was identified by staining with Congo red and examining for green birefringence under polarized light. SAA was localized immunohistochemically. Texas Red-labeled SAA was visualized in living cultures by fluorescence confocal microscopy. AEF was characterized by Western blot analysis using anti-SAA antiserum and N terminal sequence analysis. Subunits comprising amyloid in fibroblast cultures were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. RESULTS: Amyloid was produced in fibroblast cultures by an AEF-dependent mechanism. AEF, added to culture medium as insoluble protein precipitates, adhered to fibroblast monolayers. SAA bound preferentially to the adherent precipitates. Coincident with SAA binding, precipitates developed an affinity for Congo red. Over time, as more SAA was added, networks of Congo red-positive material producing bright green birefringence also developed outward from AEF precipitates. Amyloid built upon AEF in this manner was composed of full-length SAA. No amyloid was produced in cultures treated with either SAA or AEF alone. SAA and SAA peptides processed in the C-terminal region were the most prominent proteins in the glycerol-extracted AEF preparation. CONCLUSION: AEF binds to fibroblast monolayers and acts as a sink for SAA. SAA that collects on AEF assembles into an amyloid structure. Thus, it is concluded that AEF serves as both a nidus and a template for amyloid formation. PMID- 12746918 TI - Nonsteroidal antiinflammatory drugs and a selective cyclooxygenase 2 inhibitor uncouple mitochondria in intact cells. AB - OBJECTIVE: Uncoupling of isolated mitochondria by nonsteroidal antiinflammatory drugs (NSAIDs) has been considered relevant to the development of gastrointestinal (GI) side effects. We investigated the occurrence of NSAID induced uncoupling of mitochondria in intact cells (rat thymocytes) compared with the effects of a selective cyclooxygenase 2 (COX-2) inhibitor. METHODS: Oxygen consumption and mitochondrial membrane potential were simultaneously measured amperometrically and by distribution of radioactive tracer molecules, respectively, in the presence and absence of pharmacologically relevant concentrations of the NSAIDs indomethacin and diclofenac and the selective COX-2 inhibitor SC-236. Analysis of data by a technique related to top-down elasticity analysis permitted assessment of the influence of these compounds on individual components of cellular energy metabolism. RESULTS: Indomethacin, diclofenac, and SC-236 increased proton leak in isolated mitochondria. Both diclofenac and SC-236 significantly stimulated proton leak in intact cells and simultaneously inhibited substrate oxidation and ATP turnover. Oxygen consumption rates of isolated cells remained unchanged over a wide concentration range of the drugs, despite significant effects on subsystems of cellular energy metabolism. CONCLUSION: NSAIDs and selective COX-2 inhibitors have significant and equally directed effects on cellular energy metabolism. They both uncouple mitochondrial respiration and inhibit substrate oxidation and ATP turnover. However, the topical effect and selective COX-2 inhibition may not be sufficient to cause NSAID-like damage to the GI tract. PMID- 12746919 TI - Genetic basis of tissue specificity of vasculitis in MRL/lpr mice. AB - OBJECTIVE: To clarify the mode of inheritance of the tissue distribution of vasculitis in MRL/Mp-lpr/lpr (MRL/lpr) lupus-prone mice and to identify the susceptibility loci. METHODS: Vasculitis in individual MRL/lpr, C3H/HeJ-lpr/lpr (C3H/lpr), (MRL/lpr x C3H/lpr)F(1), and (MRL/lpr x C3H/lpr)F(2) intercross mice was analyzed by histopathologic grading of main branches of the aorta and of medium-sized arteries in the lower limbs. Genomic DNA samples from F(2) intercross mice were examined by simple sequence-length polymorphism analysis, and the polymorphic microsatellite markers highly associated with vasculitis in each tissue were determined as vasculitis susceptibility loci. RESULTS: A susceptibility locus with significant linkage to vasculitis of main branches of the aorta was mapped on chromosome 4 at D4Mit213 (map position 13.3cM) selectively in males, while vasculitis of medium-sized arteries in the lower limbs was mapped to different chromosomes: at D8Mit31 on chromosome 8 (map position 33.0) selectively in females and at D5Mit36 on chromosome 5 (map position 65.0). All of these were different from the previously defined loci governing susceptibility to vasculitis involving the kidneys. CONCLUSION: Systemic vasculitis in MRL/lpr mice is genetically controlled with cumulative effects of multiple gene loci, each of which has tissue specificity. PMID- 12746920 TI - CD25+ cell depletion hastens the onset of severe disease in collagen-induced arthritis. AB - OBJECTIVE: CD4+,CD25+ T regulatory cells may offer opportunities to intervene in the course of autoimmune disease. We wished to evaluate their potential for influencing systemic and chronic joint inflammation by investigating their involvement in collagen-induced arthritis (CIA). METHODS: We depleted DBA/1 mice of CD25+ regulatory cells by injection of a depleting monoclonal antibody specific for CD25 14 days before a single immunization with type II collagen (CII) in Freund's complete adjuvant. CD4+,CD25+ T cells were adoptively transferred to some groups of mice during immunization. Mice were then scored for signs of arthritis, and blood was taken periodically to measure the amounts of CII-specific antibodies. Splenocytes of treated mice were examined in vitro to determine the effects of depletion on proliferation to CII and control antigens. RESULTS: CD25+ cell-depleted DBA/1 mice had significantly more severe disease than control mice following collagen immunization. The magnified severity was also accompanied by higher antibody titers against collagen, and in vitro tests showed increased proliferation of collagen-specific T cells. Adoptively transferring CD4+,CD25+ T cells into depleted mice was shown to reverse the heightened severity. Control mice, which were depleted and immunized with the neoantigen keyhole limpet hemocyanin (KLH), had neither an increased antibody response toward KLH nor an augmented proliferative response, indicating that CD25+ cell depletion preferentially affects immunity against self antigen. CONCLUSION: These results establish a link between CD4+,CD25+ regulatory cells and CIA and provide a rationale for investigating CD4+,CD25+ T regulatory cells in the treatment and prevention of arthritis. PMID- 12746921 TI - Genetic linkage of primary hip osteoarthritis with restricted areas on chromosome 11q: comment on the article by Chapman et al. PMID- 12746924 TI - Increased mortality in early inflammatory polyarthritis: comment on the article by Goodson et al. PMID- 12746925 TI - CD34-selected versus unmanipulated grafts for severe rheumatoid arthritis: comment on the article by Moore et al. PMID- 12746926 TI - High-dose cyclophosphamide therapy without stem cell rescue for severe refractory autoimmune illnesses: comment on the article by Moore et al. PMID- 12746929 TI - Efficacy of low-dose versus high-dose cyclophosphamide in lupus nephritis: comment on the article by Houssiau et al. PMID- 12746931 TI - Persistent efficacy of tumor necrosis factor alpha blockage therapy in SAPHO syndrome: comment on the article by Wagner et al. PMID- 12746932 TI - Thrombotic thrombocytopenic purpura in a patient with Behcet's disease. PMID- 12746934 TI - Some anti-proinflammatory musings: proinflammatory is inflammatory. PMID- 12746937 TI - Noninvasive ventilation in the pediatric intensive care unit for children with acute respiratory failure. AB - Noninvasive ventilation, a novel treatment to increase alveolar ventilation, is accomplished with either subatmospheric or positive pressure administered via an external interface. In adults with acute respiratory failure, noninvasive positive pressure ventilation (NPPV) is superior to standard therapy in preventing intubation and reducing mortality. The role of NPPV in pediatric-age patients with acute respiratory distress is not as well established. Early case reports showed that NPPV treatment does acutely improve both the clinical manifestations of respiratory distress and respiratory gas exchange in children with respiratory distress. However, it is not clear whether NPPV in this setting can prevent vs. delay endotracheal intubation. Other uses of NPPV in the pediatric intensive care unit include the treatment of upper airway obstruction, atelectasis, and exacerbations of neuromuscular disorders, and to facilitate weaning from invasive mechanical ventilation. Successful use of NPPV in young infants with respiratory distress is impeded by the lack of suitable size interfaces, and the response characteristics of commercially available bilevel ventilators. Despite these challenges, NPPV is a promising alternate to standard therapies in the treatment of acute respiratory distress in the pediatric-age patient. PMID- 12746938 TI - Prevalence of symptoms of asthma and allergies in schoolchildren in Gondar town and its vicinity, northwest Ethiopia. AB - A low level of allergic disorders has been generally reported in children from developing countries. A survey was conducted to determine the prevalence of symptoms of asthma and allergies among schoolchildren in Gondar, northwest Ethiopia. A standardized self-administered questionnaire developed by the International Study of Asthma and Allergies in Children (ISAAC) was used. In total, 3,365 children aged 13-14 years were surveyed at school, with a response rate of 98.4%. The female to male ratio was 1.5:1. The 12-month-period prevalences of wheeze (16.2%), rhino-conjunctivitis (14.5%), and eczema (10.9%) were comparable to findings from Addis Ababa, but significantly higher than reported from Jimma. A striking seasonal pattern of rhinitis symptoms is observed during the months of September-December with a peak in November, corresponding with the main pollen season of the area. Overall, about 36% of the children reported symptoms of atopic disorders. However, the prevalence of diagnosed atopic diseases (8.6%) is less than a quarter of the symptom prevalence rate. This may reflect a generally poor awareness and low health service utilization, or underdiagnosis by health facilities. In conclusion, it appears that the prevalence of atopic diseases in Gondar area is high, and there is also wide variation within the country. This calls for further epidemiological studies with more objective assessments of atopy, such as allergen and bronchial hyperresponsiveness tests and ecological analysis. PMID- 12746939 TI - Oral administration of specific yolk antibodies (IgY) may prevent Pseudomonas aeruginosa infections in patients with cystic fibrosis: a phase I feasibility study. AB - Respiratory infection is the major cause of morbidity and mortality in cystic fibrosis (CF) patients. Chronic Pseudomonas aeruginosa (PA) infections ultimately occur in virtually all patients. It is impossible to eradicate PA when a patient has been chronically colonized. Immunotherapy with specific egg-yolk antibodies (IgY) may be an alternative to antibiotics for the prevention of PA infections. We wanted to determine if treatment with specific IgY can prolong the period between the first and the second PA colonization? And long-term, can the treatment diminish the number of positive PA cultures and postpone the onset of chronic colonization? CF patients gargled daily with an IgY-antibody preparation, purified from eggs of hens immunized with PA bacteria. They were compared to a group of patients who did not gargle with the preparation. Both groups had their first colonization with PA eradicated by antibiotics. The basic treatment was essentially the same in both groups. In the initial study, the period between the first and second colonization with PA was significantly prolonged for the treated vs. the control group (Kaplan-Meier P = 0.015, Breslow test). In the prolonged study, the treated group had only 2.5 sputum cultures positive for PA per 100 months of observation, and none of these patients became chronically colonized with PA. No adverse events were reported. In the control group, 13.7 cultures per 100 months of observation were positive for PA, and 5 (24%) patients became chronically colonized with PA. This feasibility study shows that antipseudomonal IgY has the potential to effectively prevent PA colonization without any severe adverse effects. A phase III study should be initiated. PMID- 12746940 TI - Correlation of six different cystic fibrosis chest radiograph scoring systems with clinical parameters. AB - In past decades, several chest radiograph scoring systems for cystic fibrosis were developed. This study was performed to compare interobserver variability of six different radiograph scores and to correlate them with clinical parameters. Thirty chest radiographs of 30 patients with cystic fibrosis were scored according to Shwachman-Kulczycki scoring, Chrispin-Norman scoring, adjusted Chrispin-Norman scoring, Brasfield scoring, Wisconsin scoring, and the Northern scoring system by two independent observers. Data on clinical parameters such as lung function, nutritional status, and infectious exacerbation rate, obtained simultaneously with the chest radiograph, were reviewed. Interobserver variability was low (Pearson's correlation coefficients, 0.76-0.84; all P < 0.01), and scores had good limits of agreement (Bland and Altman). Correlation of radiograph score with clinical parameters was good for most pulmonary function test data (correlation coefficients from 0.72-0.78 for percent of forced expired volume in 1 sec (FEV(1)%) predicted and from 0.69-0.74 for FVC% predicted) and for infectious exacerbation rate (correlation coefficients from 0.68-0.73). All six radiograph scoring systems, especially the Chrispin-Norman score, showed a low interobserver variability and correlated well with lung function tests, especially FEV(1)% predicted and infectious exacerbation rate, and moderately with maximum work capacity and thoracic mobility. PMID- 12746941 TI - Montelukast does not protect against hyperoxia-induced inhibition of alveolarization in newborn rats. AB - Impaired lung development has been demonstrated in neonatal animals exposed to hyperoxia. High lung cys-leukotriene levels may be a contributing factor towards the increase in oxygen toxicity. We investigated the effect of cysteinyl leukotriene inhibition using the receptor antagonist, montelukast (MK, Singulair), on hyperoxia-induced changes in lung parenchymal structure in neonatal rat pups. Rat pups were exposed to 21% O(2) (air) or 50% O(2) (moderate hyperoxia) from days 1-14 after birth, and were administered the cys-leukotriene receptor antagonist MK (1 mg/kg/day) or normal saline from days 4-14. Somatic growth and morphometric measurements were done on day 15. There was a significant increase in bronchoalveolar lavage fluid cysteinyl-leukotriene levels (+61.9%) when animals were exposed to hyperoxia. O(2) exposure significantly decreased the specific internal surface area by 13%. There was a nonsignificant 5.8% and 19.6% increase in mean chord length and mean alveolar diameter, respectively, as well as an 8.6% decrease in lung volume to body weight ratio. Inhibition of only one arm of the arachidonic-acid cascade by MK was not sufficient to prevent these oxygen-induced changes. PMID- 12746942 TI - Pulmonary manifestations and function tests in children genetically diagnosed with FMF. AB - Familial Mediterranean fever (FMF) is characterized by recurrent episodes of peritonitis, pleuritis, and synovitis. Among the pulmonary manifestations of FMF, pleuritis is the most common. Long-term sequelae of the respiratory system have not been described in FMF patients. We describe the pulmonary manifestations and function tests in a group of children who were found by genetic screening to be homozygous for the FMF gene. We surveyed 48 patients of Mediterranean extraction (aged 6-18 years) who were evaluated for a variety of pulmonary symptoms, and in whom clinical and genetic studies confirmed a diagnosis of FMF. All patients underwent complete pulmonary function tests, which included spirometry, body plethysmography, and single-breath carbon monoxide diffusion (Dlco). Forty percent of the Jewish patients, but only 8% of the Arab patients (P < 0.001), suffered from pulmonary manifestations during an attack of FMF. Jewish patients who were homozygotes for the M694V mutation suffered significantly more from episodes of pleuritis, cough, and rapid, shallow breathing than Arab patients, who were either homozygotes for the V726A mutation or bore any other combination of mutations. Three patients (6%) had mild restrictive lung disease, all of them homozygotes for the M694V mutation. In 3 further patients, obstructive lung impairment was found. Pulmonary manifestations during FMF attacks are significantly more common in the Jewish population bearing the M694V mutation. Restrictive lung impairment was found in a small number of these patients with a severe course of the disease; however, the series is too small to draw conclusions about long-term sequelae of the respiratory system in FMF patients. PMID- 12746943 TI - Early postnatal dexamethasone influences matrix metalloproteinase-2 and -9, and their tissue inhibitors in the developing rat lung. AB - In order to test the hypothesis that early postnatal exposure to dexamethasone (Dex) influences matrix metalloproteinases (MMP)-2 and -9, as well as their tissue inhibitors (TIMP-1 and -2) in the developing rat lung, newborn rats (3 litters/group) were treated with low Dex (0.1 mg/kg/day, IM), high Dex (0.5 mg/kg/day), or equivalent volumes of saline at 5 days postnatal age (P5), P6, and P7. Lung weight and lung MMP and TIMP levels were determined at sacrifice (7 days postinjection, P14; at weaning, P21; and at adolescence, P45, n = 10/group and time). Dex did not adversely affect lung weight or lung MMP-2 levels, which peaked in all groups at P21 and then fell by P45. In contrast, Dex decreased TIMP 2 at all time intervals, but achieved statistical significance only at P45. An imbalance in MMP-2/TIMP-2 ratio was noted at P21, with elevations occurring in the low and high Dex-treated groups. Lung MMP-9 levels remained comparable with controls during low Dex treatment. However, high Dex exposure resulted in elevated lung MMP-9 levels at P21 and P45. Lung TIMP-1 levels increased only with high Dex exposure at P14 and P21, whereas the lung MMP-9/TIMP-1 ratio was elevated at P21 in the high Dex group, and at P45 in both Dex-treated groups. These data provide evidence that early postnatal dexamethasone results in an imbalance between gelatinase-A and -B, and their tissue inhibitors in the developing rat lung. These changes may be responsible, in part, for some of the known maturational effects of steroids on lung structure in the newborn. PMID- 12746944 TI - Clinical application of direct sputum sensitivity testing in a severe infective exacerbation of cystic fibrosis. AB - A 16-year-old male with cystic fibrosis (CF) was admitted to hospital with a severe infective exacerbation. Despite standard management, including conventionally selected intravenous antibiotics for Pseudomonas aeruginosa, chest physiotherapy, and institution of noninvasive ventilation (NIV) for progressive hypercapneic respiratory failure, he continued to deteriorate. Direct sputum sensitivity testing (DSST) revealed a novel combination of antibiotics that resulted in a rapid and remarkable clinical improvement. DSST is a form of "whole" sputum sensitivity testing that provides information on antibiotic synergy, and may more accurately reflect in vivo antibiotic sensitivity patterns in cystic fibrosis. PMID- 12746945 TI - Determination of maximal voluntary ventilation in children with cystic fibrosis. AB - Maximal voluntary ventilation (MVV) may be determined directly by the sprint method or calculated from pulmonary function data, using the functions MVV = forced expired volume in 1 sec (FEV(1)) x 35 or MVV = FEV(1) x 40. The purpose of this paper was to test the validity of the equation over a wide range of lung function in children. Cystic fibrosis (CF), a chronic lung disease where children typically have a wide range of pulmonary function, was chosen as the study requirement. Spirometric data from 332 children with CF who underwent pulmonary function testing between 1987-2000 were stratified according to disease severity, and box-plots comparing the ratio of MVV to FEV(1) for each category were generated. As results indicated that the equation underestimates true MVV proportionally to the degree of airflow limitation, a new function to predict MVV for this population was derived and tested. The new equation was derived using data from patients who were tested on odd-numbered days (group A). The validity of the new equation was then tested on the patients tested on even-numbered days (group B). To test its validity, the results were compared to the "gold standard" sprint values using a Bland and Altman plot. MVV was expressed as a function of FEV(1) and predicted FEV(1): MVV = 27.7(FEV(1)) + 8.8(PredFEV(1)) (R(2) = 0.98, P < 0.05). In this way, the accuracy of the new equation was confirmed. Whenever possible, we recommend MVV be determined by the sprint method in accordance with ATS guidelines. If this is not feasible, we recommend considering the new prediction equation. PMID- 12746946 TI - Teaching of pulmonary auscultation in pediatrics: a nationwide survey of all U.S. accredited residencies. AB - Structured teaching of pulmonary auscultation is greatly underrepresented during internal medicine (IM) or family practice (FP) training. It is not known, however, whether this underrepresentation applies to the other major field of primary care, pediatrics. To answer this question, we surveyed all accredited U.S. residencies in pediatrics by mailing a 1-page questionnaire to 174 pediatrics program directors, and by comparing results to those previously gathered from internal medicine and family practice residencies. Pediatrics directors' response rate was 62%. More pediatrics than family practice residencies offered structured teaching of pulmonary auscultation (21.5% vs. 9.7%, P < 0.004). When compared to internal medicine programs, this difference showed a trend toward significance (21.5% for pediatrics and 14.1%, for internal medicine, P = 0.08). Teaching modalities included: lectures (91.2%); audiotapes (13%); seminars (8.3%); and miscellaneous (21.7%). University-affiliated residencies taught auscultation significantly more frequently than nonuniversity affiliated programs (25.4% vs.10.5%, P = 0.07). Pediatrics directors gave great importance to pulmonary auscultation, and wished for more time devoted to its teaching (5.52 +/- 0.84 and 5.01 +/- 1.07, respectively, on a 1-6 scale, with 6 indicating the highest value). They also attributed great clinical importance to 13 commonly encountered pulmonary auscultatory events (all rated, on average, between 4-5.8 on a 1-6 scale, with 6 indicating highest importance). In summary, training programs in pediatrics offered significantly more structured teaching of pulmonary auscultation than IM or FP residencies. Whether this difference in teaching may have a beneficial impact on the auscultatory proficiency of pediatric residents, as compared to internal medicine and family practice trainees, needs to be determined. PMID- 12746947 TI - Non-CF bronchiectasis: clinical and HRCT evaluation. AB - Childhood bronchiectasis not related to underlying disease is still common in some populations in affluent countries. The aims of the study were to: 1) describe demographics, 2) evaluate the effectiveness of routine investigations, and 3) determine the relationship between spirometry and radiology scoring systems, in children with chronic suppurative lung disease (CSLD) living in Central Australia. Data of children living in Central Australia aged 70%) and early hospitalisation for pneumonia were common (median age, 0.5 years). Previous admissions for pneumonia were almost universally present and significantly more common than bronchiolitis (95% CI for proportional difference, 0.4-0.51). Although the majority did not have a treatable underlying cause, investigations had significant impact on management in 12.3% of children. None of the chest HRCT scores related to any spirometry data. In conclusion, CSLD is unacceptably common in indigenous children of this region, commences early in life, and is associated with significant comorbidities. Spirometry data do not reflect the severity of lung disease in HRCT scans. While improvement in the living standards is of utmost importance, the medical management that includes thorough investigations of these children should not be neglected. PMID- 12746949 TI - Iatrogenic pleuropulmonary charcoal instillation in a teenager. AB - Activated charcoal given through a nasogastric tube is a standard intervention for many types of toxic ingestions in the emergency department. This case study describes a teenage girl whose multidrug overdose was complicated by accidental charcoal instillation into her left lung and pleural space through a misplaced nasogastric tube. The ensuing empyema did not respond to antibiotic therapy alone, probably due to the inherent properties of charcoal, and required a chest tube placement with continuous irrigation. Unlike previously reported cases, this patient did well clinically, without long-term morbidity. PMID- 12746951 TI - [Endocrine diseases: progress in their diagnosis and treatment]. PMID- 12746950 TI - Rapid progression of intrapulmonary arteriovenous shunting in polysplenia syndrome associated with biliary atresia. AB - This report describes a patient with biliary atresia (BA) associated with polysplenia syndrome who showed a rapid progression of intrapulmonary arteriovenous shunting (IPS), resulting in a fatal outcome. Intrauterine ultrasonography at 36 weeks of gestation revealed fetal abnormalities, including situs inversus, absent retrohepatic inferior vena cava, and azygous connection. She was diagnosed postnatally as BA because of persistent acholic stool and neonatal jaundice. She underwent hepatic portoenterostomy at age 158 days. The gallbladder and the hepatic ducts were hypoplastic, and the common bile duct was absent. Magnetic resonance image and operative findings also identified polysplenia and an absent portal trunk. Liver histology showed cirrhotic changes and bile duct proliferation. Postoperatively, she achieved good bile secretion, with gradual decrease of total bilirubin. However, she had repeated febrile episodes, and computerized tomography at age 7 months showed multiple liver cysts. Thereafter, she presented with exertional dyspnea. Contrast-enhanced echocardiography showed IPS with a degree of 2/III at age 8 months and 3/III at 10 months. (99m)Technetium-labeled macroaggregated albumin ((99m)Tc-MAA) scintigraphy revealed a shunt ratio of 25.5% at 9 months and 39.7% at 10 months. Percutaneous transhepatic drainage of the bile cysts was performed without success. Sludged bile was obtained. However, respiratory distress rapidly progressed, and she died at age 11 months. In the present patient, the association of polysplenia syndrome and absent portal vein with BA, as well as liver cirrhosis, seemed to be contributing factors to rapid progression of IPS in early life. PMID- 12746953 TI - [From diagnosis to treatment: the posterior pituitary gland]. PMID- 12746952 TI - [From diagnosis to treatment: The anterior pituitary gland]. PMID- 12746954 TI - [From diagnosis to treatment: the thyroid gland]. PMID- 12746948 TI - Palivizumab prophylaxis of respiratory syncytial virus disease in 2000-2001: results from The Palivizumab Outcomes Registry. AB - The objective of the Registry was to characterize the population of infants receiving prophylaxis for respiratory syncytial virus (RSV) disease by describing the patterns and scope of usage of palivizumab in a cross section of US infants. RSV hospitalization outcomes were also described. The Palivizumab (Synagis, MedImmune, Inc., 25 West Watkins Mill Road, Gaithersburg, MD 20878) Outcomes Registry was a prospective multicenter survey conducted at 63 sites. Demographics, injection history, and RSV hospitalization outcomes were collected on 2,116 infants receiving palivizumab. Infants were enrolled in the Registry between September 1, 2000-March 1, 2001, at the time of their first injection. Infants born at less than 32 weeks of gestation accounted for 47% of infants enrolled, and those between 32-35 weeks accounted for 45%; approximately 8% were greater than 35 weeks of gestation. Lower RSV hospitalization rates were observed in infants who had greater adherence to regularly scheduled injections. Nearly one-half of all hospitalizations occurred within the first and second injection intervals, suggesting the importance of early RSV protection. The confirmed RSV hospitalization rate of all infants in the Registry was 2.9%; the rate was 5.8% in infants with chronic lung disease of infancy, and 2.1% in premature infants without chronic lung disease. In conclusion, these data support the continued effectiveness of palivizumab prophylaxis for severe RSV lower respiratory tract disease in a large cohort of high-risk infants from geographically diverse pediatric offices and clinics. The Palivizumab Outcomes Registry provides an opportunity to assess palivizumab utilization and clinical effectiveness in the US. PMID- 12746955 TI - [From diagnosis to treatment: the parathyroid gland and abnormal calcium metabolism]. PMID- 12746957 TI - [From diagnosis to treatment: the sex glands]. PMID- 12746956 TI - [From diagnosis to treatment: the adrenal gland]. PMID- 12746958 TI - [From diagnosis to treatment: pancreatic endocrine tumor]. PMID- 12746959 TI - [From diagnosis to treatment: multiple endocrine neoplasia (MEN)]. PMID- 12746960 TI - [Current topics in endocrinology: ghrelin]. PMID- 12746961 TI - [Current topics in endocrinology: adipocytokines]. PMID- 12746962 TI - [Current topics in endocrinology: incretin]. PMID- 12746963 TI - [Current topics in endocrinology: transcription factors and endocrine diseases]. PMID- 12746964 TI - [Current topics in endocrinology: blood vessels and bones]. PMID- 12746965 TI - [For more accurate diagnosis of endocrine diseases: discussion]. PMID- 12746966 TI - [Non-autoimmune fulminant type 1 diabetes mellitus presenting with diabetic ketoacidosis after the delivery]. PMID- 12746968 TI - [Crohn's disease presenting arthritis of knees without any gastrointestinal symptoms]. PMID- 12746967 TI - [Destructive thyroiditis with elevated thyroid stimulation blocking antibody]. PMID- 12746970 TI - [Bioterrorism--as a source of infection in clinical medicine]. PMID- 12746969 TI - [Involvement of neuromuscular disorders in maternally inherited diabetes with deafness (MIDD)]. PMID- 12746971 TI - [Mini-transplantation]. PMID- 12746972 TI - [The concept of bone marrow transplantation in autoimmune disease]. PMID- 12746973 TI - SARS hit home. PMID- 12746974 TI - Malignancy-associated changes in lactiferous duct epithelium. AB - OBJECTIVE: To determine whether cells from histologically normal appearing epithelium of the lactiferous duct from women with a remote ductal lesion in the breast provide any clues indicating the existence of such a lesion. STUDY DESIGN: Tissue sections cut to 4 microns and stained with hematoxylin and eosin were prepared from duct tissue of 20 women with breast lesions and of 20 women free of any such lesion who had undergone mammoplastic procedures or resection for benign reasons. One hundred nuclei were measured from each case. Measures of nuclear deviation from normal were computed, discriminant functions were derived, and multivariate significance tests were conducted. RESULTS: Nuclei from histologically normal appearing regions of lactiferous duct epithelium from women harboring distant lesions exhibited changes in the distribution pattern of their nuclear chromatin, indicating the presence of these lesions. The statistical significance of these changes was documented. The changes were clearly evident in all 20 subjects with lesions and were not observed in 19 of the 20 subjects without lesions. CONCLUSION: The results suggest that studies aimed at detecting malignancy-associated changes in cells collected by ductal lavage might lead to a minimally invasive screening procedure for breast lesions. PMID- 12746975 TI - Fine needle aspiration biopsy of the breast. Value of nuclear morphometry after different sampling methods. AB - OBJECTIVE: To study the potential of nuclear morphometry in supporting the interpretation of fine needle aspiration biopsy (FNAB) samples of the breast fixed in 50% ethanol and centrifuged on slides. STUDY DESIGN: Computerized morphometry was used to outline the nuclei of breast epithelial cells in breast cancer, fibroadenoma and fibrocystic disease. The diagnoses were histologically confirmed. We applied 2 different sampling methods (measurements done on cell groups and on free cells). RESULTS: The mean nuclear area of cell groups of malignant samples (23) varied from 42 to 125 microns 2, in fibroadenomas from 30 to 50 microns 2 and in fibrocystic disease from 26 to 57 microns 2. The mean nuclear area of free cells varied as follows: cancer, 66-181 microns 2; fibroadenoma, 33-70 microns 2; fibrocystic disease, 35-60 microns 2. Apocrine metaplasia was excluded from comparison on a morphologic basis. CONCLUSION: The study suggests that if the mean nuclear area of cell groups is < 42 microns 2, the lesion is probably benign; if > 57 microns 2, and apocrine metaplasia is excluded, malignancy should be considered. The differential diagnosis between carcinoma and fibroadenoma could be based on free cells: mean area of free cell nuclei < or = 65 microns 2 suggested a benign lesion, and of > or = 71 microns 2 suggested a malignant lesion. Morphometric nuclear size features (exemplified by nuclear area) appeared efficient in distinguishing between malignant and benign lesions when measured from free cells and cell groups. PMID- 12746976 TI - Diagnostic use of mean nuclear area and nuclear DNA content in preoperative breast cancer cytology. AB - OBJECTIVE: Fine needle aspiration (FNA) cytology of breast lumps is a routine procedure, and the diagnostic accuracy can be 95%. Occasional discrepancies arise, and it would be valuable to have additional parameters for accurate diagnosis. We evaluated nuclear DNA content and mean nuclear area (MNA) using image cytometry in the diagnosis of preoperative breast cancers by FNA in those with a discrepancy between clinical, radiologic and cytologic diagnoses. STUDY DESIGN: One hundred eighteen consecutive preoperative FNA samples were evaluated for nuclear DNA and MNA and were compared to cytologic and postoperative histologic diagnoses. RESULTS: Sensitivity, accuracy and positive predictive value of routine cytology were 95%, 90%, 95% as compared to nuclear DNA (66%, 66%, 96%) and MNA (61%, 61%, 97%). Combining these 3 parameters gave a sensitivity of 97%, accuracy of 94% and positive predictive value of 99%. CONCLUSION: These results demonstrate that nuclear DNA and MNA combined with routine cytology may be useful adjuncts in preoperative breast cancer cytologic diagnosis when discrepancies arise. This may lead to better and more accurate planning of treatment regimens in preoperative breast cancer patients. PMID- 12746977 TI - Nuclear morphometric features of epithelial cells lining keratocysts. AB - OBJECTIVE: To investigate the nuclear morphometric features of epithelial cells lining keratocysts and some other odontogenic cysts. STUDY DESIGN: All cases were selected from the archives of the Department of Pathology, Gulhane Military Medical Academy, as follows: 20 keratocysts and 10 dentigerous and 10 radicular cysts. Nuclear morphometric variables were measured on hematoxylin and eosin stained histologic slides. Basal and intermediate cells of the epithelium were evaluated separately. Nuclei of the cells were outlined interactively and measured using a specially written macro program. Area, feret ratio (ratio of the longest nuclear axis to the shortest one) and circularity (F circle) of the nuclei were calculated. Additionally, nuclear densitometric analysis was performed on the keratocyst cases. RESULTS: The number of cells in the basal layer (cell density) was higher in keratocysts than in other cysts. The mean nuclear area of basal cells was smaller than of intermediate cells in both keratocysts and other cysts (P < .001). The feret ratio values revealed that basal cell nuclei of keratocysts were more ovoid as compared to those of other cysts (P < .001). Nuclear densitometric findings showed that the DNA indices of all keratocyst cases were close to 1.0, and the cells were considered diploid. CONCLUSION: Increased cell density, a more ovoid nuclear shape and more variation in the size of basal layer cell nuclei in keratocysts were helpful in differentiating these lesions from other odontogenic cysts. PMID- 12746979 TI - A systematic approach to multispecies sperm morphometric characterization. AB - OBJECTIVE: To investigate the morphologic variety of sperm heads between several species through a comprehensive set of measurements, some of which are presented here for the first time, and to perform cluster analysis. STUDY DESIGN: Fifty images were obtained for each species, preprocessed, normalized and characterized through a series of morphologic measures. All possible 2 x 2 combinations of measurements were assessed in terms of objective (discriminant analysis) and subjective (visual inspection) methods. RESULTS: The specific potential of each measure and 2 x 2 combinations were identified with respect to 2 main situations: separation between species and separation of alterations within a species, possibly corresponding to anomalies or subpopulations. Several of the measurements led to reasonable separation between species, and some specific measurements allowed the identification of morphologic alterations. CONCLUSION: The potential of relatively more sophisticated measurements and concepts for sperm analysis was corroborated. Although most of the measures considered were observed to allow good separation between species, some of the measurements, such as hydrodynamic coefficients, anterior-posterior and side symmetry, shape factor and first Fourier descriptor, accounted for the identification of secondary clusters within specific species, possibly indicating pathologic alterations or signaling the presence of subpopulations. PMID- 12746978 TI - Analysis of the reliability of manual and automated immunohistochemical staining procedures. A pilot study. AB - OBJECTIVE: To study the variation in the number of stained cells and staining intensity comparing 2 immunostainers and manual staining for estrogen receptor (ER) expression in breast carcinoma. STUDY DESIGN: In 5 cases, 15 consecutive paraffin sections were investigated after simultaneous immunohistochemical ER staining. The slides were evaluated using a CM-2 TV image analysis system (Hund, Wetzlar, Germany). One viewing field, identified around a histologic structure present on all 15 sections, was analyzed. The percentage of immunoreactive cells (PP), mean grey values of the immunopositive (GVpos.) and immunonegative nuclei (GVneg.), and immunohistochemical staining intensity (SI, defined as GVneg. GVpos.) were calculated. RESULTS: The mean PP values were higher for immunostainers A (70.2%) and B (53.8%) than for manual staining (40.8%). The results were significantly different comparing the 2 immunostainers (P = .0143) or immunostainer A and manual staining (P < .0001). Also, the mean SI values were higher for immunostainers A (24.5 +/- 2.8% [CV]) and B (18.5 +/- 31.1%) than for manual staining (10.8 +/- 33.8%). These differences revealed statistical significance comparing the immunostainers with manual staining (.0001 < P = .0048). CONCLUSION: Our results underline the higher staining quality using immunostainers in comparison with manual staining. PMID- 12746980 TI - Digital karyometry in pancreatic adenocarcinoma. AB - OBJECTIVE: To characterize nuclei from pancreatic adenocarcinoma and nonneoplastic pancreatic tissue by digital karyometry, demonstrating specific nuclear signatures for each of them. STUDY DESIGN: Of cells from malignant and nonmalignant pancreatic tissue, 1,300 nuclei were assessed by digital karyometry from paraffin blocks stored at the Pathology Service of Hospital de Clinicas de Porto Alegre. A set of 40 features descriptive of the spatial and statistical distribution of nuclear chromatin was computed for each nucleus. Signatures were created for both types of tissue, and a distance metric from "normal" was defined and calculated for them. RESULTS: There were significant differences in 11 features between the 2 groups, allowing the creation of digital signatures. CONCLUSION: Nuclear chromatin texture signature can offer a specific digital characterization for both pancreatic adenocarcinoma and nonmalignant pancreatic tissue. Several isolated nuclear features serve as markers for the diagnosis of pancreatic adenocarcinoma. The present karyometric study of normal and malignant pancreatic tissue may be of use as a continuing tool to early diagnosis of pancreatic adenocarcinoma as it can be applied to cytologic specimens, also. In the future, studies using this technique should assess the chemopreventive potential of different agents as well as prognosis and treatment options for pancreatic adenocarcinoma. PMID- 12746981 TI - Daily workload guidelines for cytotechnologists utilizing automated, assisted screening technologies. PMID- 12746982 TI - Body mass index--specific weight gains associated with optimal birth weights in twin pregnancies. AB - OBJECTIVE: To formulate maternal weight gain guidelines, by maternal pregravid body mass index (BMI) status, associated with optimal fetal growth and birth weight in twins. STUDY DESIGN: This historical cohort study was based on 2,324 pregnancies with nonanomalous, liveborn twins (4,684 infants) from Ann Arbor, Charleston, Baltimore and Miami. Rates of maternal weight gain and fetal growth were modeled using multiple regression for 0-20 weeks, 20-28 weeks and 28-38 weeks (projected as necessary), controlling for potentially confounding factors. Optimal rates of fetal growth were defined as growth between the singleton and twin 50th percentiles, and optimal birth weights were defined as between the singleton 50th percentile and twin 90th percentile at > or = 36 weeks (2,850 2,950 g). RESULTS: Optimal rates of fetal growth and birth weights were associated with rates of maternal weight gain for underweight women of 1.25-1.75 lb/wk (0.57-0.79 kg/wk) to 20 weeks, 1.50-1.75 lb/wk (0.68-0.79 kg/wk) between 20 and 28 weeks and 1.25 lb/wk (0.57 kg/wk) from 28 weeks to delivery; for normal weight women, 1-1.5 lb/wk (0.45-0.68 kg/wk) to 20 weeks, 1.25-1.75 lb/wk (0.57 0.79 kg/wk) between 20 and 28 weeks and 1.0 lb/wk (0.45 kg/wk) from 28 weeks to delivery; for overweight women, 1-1.25 lb/wk (0.45-0.57 kg/wk) to 20 weeks, 1-1.5 lb/wk (0.45-0.68 kg/wk) between 20 and 28 weeks and 1 lb/wk (0.45 kg/wk) from 28 weeks to delivery; for obese women, 0.75-1 lb/wk (0.34-0.45 kg/wk) to 20 weeks, 0.75-1.25 lb/wk (0.34-0.57 kg/wk) between 20 and 28 weeks and 0.75 lb/wk (0.34 kg/wk) from 28 weeks to delivery. CONCLUSION: Optimal rates of fetal growth and birth weights in twins are achieved at rates of maternal weight gain that vary by period of gestation and maternal pregravid BMI status. PMID- 12746983 TI - Serum androgen markers in preeclampsia. AB - OBJECTIVE: To determine whether maternal serum levels of androgens are associated with preeclampsia in primigravid women. STUDY DESIGN: A case-control study of primigravid women with singleton pregnancies. Women diagnosed with preeclampsia (n = 15) were matched with normotensive controls (n = 30) for age and gestational age. Serum testosterone, sex hormone binding globulin, estradiol and dehydroepiandrosterone sulfate were measured before delivery. The study had 80% power to detect a 30% difference in mean testosterone concentration between cases and controls using a two-tailed test and alpha level of .05. The Student t test, Mann-Whitney U test, Wilcoxon signed ranks test and chi 2 analysis of proportions were used for analysis. RESULTS: Cases and controls did not differ in maternal age, gestational age, body mass index, tobacco use or neonatal sex. As compared with normotensive controls, preeclamptic women exhibited no statistically significant differences in median levels of total testosterone, free androgen index, sex hormone binding globulin, estradiol or dehydroepiandrosterone sulfate. CONCLUSION: Maternal serum levels of androgens do not exhibit an association with preeclampsia in primigravid women. PMID- 12746984 TI - Viral studies on amniotic fluid from fetuses with and without abnormalities detected by prenatal sonography. AB - OBJECTIVE: To study the prevalence of viruses (cytomegalovirus [CMV] adenoviruses and enteroviruses) in amniotic fluid samples from fetuses with and without anomalies detected by prenatal sonography. STUDY DESIGN: Fluid samples obtained aseptically from 474 women undergoing genetic amniocentesis at our institutions from 1995 to 1996 were stored at -20 degrees C. Fetal anomalies (renal, central nervous system, gastrointestinal and cardiac) were detected by ultrasound in 162 of the fetuses. At a later date, the samples were retrieved, blinded, and tested by virus isolation techniques for CMV, adenoviruses and enteroviruses. Fisher's exact test was used for statistical analysis. RESULTS: The prevalence of viral isolation in amniotic fluid samples in fetuses with anomalies was 2.5% for CMV, 1.3% for adenovirus and 1.2% for enterovirus. Structurally normal fetuses had prevalences of 0.3%, 0% and 0%, respectively. CONCLUSION: The prevalence of viruses, especially CMV, appears to be higher in amniotic fluid from fetuses with sonographically detected anomalies. PMID- 12746985 TI - Nonisthmic communicating uteri. Report of 3 new types in a new subclass of communicating uteri. AB - OBJECTIVE: To describe 3 new malformations and propose a new subclass of malformations, nonisthmic communicating uteri. STUDY DESIGN: Review of 17,511 hysterosalpingograms revealed 19 cases of communicating uteri. Three cases were identified that showed nonisthmic communications, previously unreported. RESULTS: The 3 new types of communicating uteri were (1) bicornuate uterus with 2 sites of communication, midcervical and isthmic; (2) septate uterus with a midcorporeal communication; and (3) bicornuate uterus with a low cervical communication and atretic hemicervix. CONCLUSION: The 3 new types of nonisthmic communicating uteri constitute a new subclass of communicating uteri. We are aware of only 1 other reported type of nonisthmic communicating uteri. Nonisthmic communications may arise from a different mechanism than isthmic communications. PMID- 12746986 TI - Management of the perineum during forceps delivery. Association of episiotomy with the frequency and severity of perineal trauma in women undergoing forceps delivery. AB - OBJECTIVE: To examine the association of the frequency and severity of perineal trauma with episiotomy performed at forceps delivery. STUDY DESIGN: This retrospective study analyzed all forceps deliveries at the Semmelweis Women's Hospital Vienna between February 1999 and July 1999. Evaluation of a possible association of episiotomy with the frequency and severity of perineal trauma was the main objective of the study. Episiotomy was not performed routinely and was either midline or mediolateral. RESULTS: In conjunction with forceps delivery episiotomy, 76/87 women (87%) underwent forceps delivery episiotomy; among those, 49/76 (64%) had a mediolateral episiotomy and 27/76 (36%) a midline episiotomy. The frequency and severity of perineal tears were significantly lower in forceps deliveries when an episiotomy was performed. When analyzing the type of episiotomy, the data revealed a statistically significantly lower frequency of perineal trauma when mediolateral episiotomy was performed as compared to midline episiotomy. CONCLUSION: If obstetric indications necessitate forceps delivery, performance of an episiotomy decreases the risk of perineal tears of all degrees. When analyzing the type of episiotomy, mediolateral episiotomy seems to be more protective against perineal trauma in women undergoing forceps delivery. PMID- 12746987 TI - Preventing ovarian hyperstimulation syndrome by inhibiting the effects of vascular endothelial growth factor. AB - OBJECTIVE: To evaluate whether inhibiting the effects of vascular endothelial growth factor can prevent the ovarian hyperstimulation syndrome. STUDY DESIGN: Rates were hyperstimulated with follicle-stimulating hormone injections. On the final day of stimulation, the rats were randomized to receive or not receive exogenous soluble fmslike tyrosine kinase (sFlt-1). Forty-eight hours later capillary permeability was determined by measuring the concentration of Evans blue dye (EB) in peritoneal irrigation fluid 3 minutes after an intravenous injection of EB. RESULTS: The peritoneal EB level was statistically significantly lower in the hyperstimulated group, which received the sFlt-1, than in the stimulation-only group. CONCLUSION: sFlt-1 may play a therapeutic role in management of the ovarian hyperstimulation syndrome by specifically inhibiting the effects of vascular endothelial growth factor. PMID- 12746988 TI - A clinical pathway for laparoscopically assisted vaginal hysterectomy. Impact on costs and clinical outcome. AB - OBJECTIVE: To evaluate the cost of and clinical outcome of implementing a clinical pathway for laparoscopically assisted vaginal hysterectomy. STUDY DESIGN: A retrospective study of the case records of patients who underwent laparoscopically assisted vaginal hysterectomy before (May-December 1997) and after (January 1998-March 1999) implementation of a clinical pathway. Data regarding resource consumption and clinical outcome represented by 10 clinical indicators were collected. Student's t test and the chi 2 test were used, as appropriate. Statistical significance was set at P = .05. RESULTS: After implementation of the laparoscopically assisted vaginal hysterectomy clinical pathway, the average total fee decreased significantly, by 8.1% (P = .03), the average inpatient drug fee decreased by 50.6% (P < .01), and the laboratory fee dropped by 56.2% (P < .01). Furthermore, the length of hospital stay significantly decreased, from 6.90 to 4.08 days (P < .01); the average operation time decreased by 24.8% (P < .01); and the average anesthesia time decreased by 21.6% (P < .01). The pre-clinical pathway and post-clinical pathway complication rate did not differ statistically, but the rate of initiating intravenous antibiotic injections > 48 hours following surgery decreased by 76.2% in the clinical pathway group (P = .02). CONCLUSION: Implementation of a clinical pathway for laparoscopically assisted vaginal hysterectomy can improve health care outcomes by decreasing length of hospital stay and admission fees and by maintaining quality of care. The clinical pathway is a good policy for maintaining cost containment and high-quality patient care. PMID- 12746989 TI - Three-dimensional yolk and gestational sac volume. A prospective study of prognostic value. AB - OBJECTIVE: To evaluate the relative prognostic value of ultrasound findings during the first-trimester scan by univariate and logistic regression analysis in a group of asymptomatic women. STUDY DESIGN: A total of 125 asymptomatic pregnant women with a singleton conceptus, concordance between menstrual age and crown rump length, and documented fetal activity, 25 for each gestational age between 6 and 10 weeks, were enrolled in the study and underwent a transvaginal sonographic examination. The following data were collected: yolk sac mean diameter and volume, gestational sac mean diameter and volume, fetal heart rate, maternal age, gestational age and presence of a retrochorial hematoma. The outcome variable was abortion, defined as pregnancy loss at any time up to 20 weeks' gestation. Normograms were constructed for volumes, mean diameters and fetal heart rate. Receiver-operator characteristic curves were performed in order to dichotomize maternal and gestational age. Univeriate analysis was performed by Fisher's exact test. Logistic regression was performed to test the relationship between independent variables and pregnancy outcome. RESULTS: In univariate analysis, the variables significantly associated with spontaneous abortion were maternal age > 34 years, yolk sac volume outside the 5th to 95th percentile, gestational sac volume < the 5th percentile and fetal heart rate outside the 5th to 95th percentile. In regression analysis only maternal age > 34 years, gestational sac mean diameter < 5th percentile and fetal heart rate outside the 5th to 95th percentile were significant in predicting abortion. CONCLUSION: Our data suggest that new three-dimensional parameters are of no clinical benefit in the prediction of abortion in nonbleeding, first-trimester pregnancy, when conventional sonographic parameters are used. PMID- 12746990 TI - Infant mortality among twins born to teenagers in the United States. Black-white disparity. AB - OBJECTIVE: To determine the magnitude of the disparity in infant mortality between twins born to black and white teenagers in the United States. STUDY DESIGN: Analysis was performed on twins born to adolescents in the United States within the period 1995-1997. The generalized estimating equations framework was used to generate relative risks after capturing the effects of sibling correlations within twin pairs. RESULTS: Infant mortality was 20% higher among black twins as compared to their white counterparts (adjusted OR = 1.20, 95% CI = 1.04-1.39). The black-white disparity in infant mortality occurred exclusively in the neonatal period (adjusted OR = 1.31, 95% CI = 1.11-1.54), with postneonatal estimates comparable (adjusted OR = 0.86, 95% CI = .63-1.17). The higher proportion of low-birth-weight infants--more specifically, those small for gestational age as opposed to preterm--among black twins was the most likely explanation for the lower survival probability among twins born to black teenagers. CONCLUSION: Black-white disparity in infant mortality among twins occurred exclusively during the first 28 days of life rather than throughout infancy. Efforts to bridge the gap should be focused on this critical period and should preferentially target those twins who are small for gestational age. PMID- 12746991 TI - Percutaneous transcatheter angiographic embolization in the management of obstetric hemorrhage. AB - OBJECTIVE: To describe the results of percutaneous arterial embolization in the management of obstetric hemorrhage. STUDY DESIGN: From February 1992 to May 2000, 33 patients with pregnancy-related hemorrhage underwent angiographic embolization to control the hemorrhage. In all cases, hemostatic embolization was performed because of intractable hemorrhage unresponsive to conservative management. All available hospital records were reviewed and detailed, and the clinical data related to complications, clinical status, estimated blood loss and blood replacement requirements, length of time of procedure, emboli used, complications associated with the procedure and results obtained were analyzed. RESULTS: Successful embolization was achieved in 31 of 33 patients. No major complications related to embolization occurred, and 3 women subsequently became pregnant. CONCLUSION: Angiographic embolization is a safe and effective method of controlling pregnancy-related hemorrhagic complications unresponsive to conservative management and maintains reproductive ability. PMID- 12746992 TI - Symphysis-fundus height and weight gain pattern in Japanese women with twin pregnancies. AB - OBJECTIVE: To determine the normogram for symphysis-fundus height and weight gain pattern in Japanese women with twin pregnancies and to examine whether fundus height and weight gain pattern are altered in women who give birth to twin infants complicated by fetal growth restriction (FGR). STUDY DESIGN: A retrospective review of the medical records of 186 twin gestations that had been followed at our institution together with 229 singleton pregnancies as a control. We examined maternal symphysis-fundus height, body weight gain and increase in body mass index (BMI) according to gestational age and compared them between women with and without FGR. RESULTS: As compared to women with singleton pregnancies, those with twins exhibited larger fundus height, larger weight gain and larger increase in BMI, all of which were observed from as early as 16-18 weeks of gestation until delivery. Mothers of twins with at least one FGR infant showed smaller fundus height, smaller weight gain and smaller increase in BMI as compared to those without. CONCLUSION: We constructed the normogram for symphysis fundus height and weight gain pattern in Japanese twin gestations and determined the characteristic change in these parameters in FGR in women with twins. These results may be of use in identifying mothers of twins with FGR. PMID- 12746993 TI - Primary extrarenal rhabdoid tumor of the ovary. A case report. AB - BACKGROUND: Malignant rhabdoid tumors are rare, aggressive neoplasms that consist of both renal and extrarenal subtypes. Although extrarenal rhabdoid tumors have been documented at multiple extrarenal sites, to our knowledge no primary ovarian cases have been reported. CASE: An 18-year-old, Caucasian woman was diagnosed with a pure primary extrarenal rhabdoid tumor of the ovary following diagnostic laparoscopy for pelvic pain. The tumor exhibited rapid growth, failed to respond to chemotherapy and led rapidly to death. CONCLUSION: Although no other reports on primary ovarian extrarenal rhabdoid tumor have been published, the aggressive behavior of the tumor in this patient was similar to that seen in patients with metastatic disease. PMID- 12746994 TI - Angiography as treatment for a high cervical tear. A case report. AB - BACKGROUND: High cervical tears are a rare but serious complication of second trimester induced abortion. Angiographic embolization can successfully treat selected cases. This is the first case of serious hemorrhage successfully treated by angiography alone, without mechanical occlusion, after failed surgical repair. CASE: A 22-year-old primigravida sustained a high cervical tear during dilation and evacuation (D&E) abortion at 21 weeks' gestation. After stabilization with 4 units of packed red cells and vaginal packing, attempted surgical repair failed. The following day, angiography alone pinpointed and stanched the small arteriolar branch of the uterine artery responsible for the bleeding. CONCLUSION: Injury to a tiny arteriolar branch of the uterine artery during D&E abortion may be impossible to palpate or visualize, and hemostatic sutures placed extrinsically on the cervical mucosa may not be therapeutic. On occasion, injection of contrast material, without mechanical occlusive devices, may be corrective. PMID- 12746995 TI - Rupture of a uterine horn after laparoscopic salpingectomy. A case report. AB - BACKGROUND: Uterine rupture after salpingectomy, especially associated with cornual resection, is a rare, serious pregnancy complication. CASE: A spontaneous uterine rupture occurred during the second trimester of pregnancy, following salpingectomy with resection of the interstitial portion. Conservative treatment was performed, and fertility was preserved. CONCLUSION: Postsalpingectomy pregnancies must be carefully and frequently monitored, with ultrasonography used at the slightest clinical symptom. A postsalpingectomy rupture must be treated surgically, preferably with conservative treatment rather than hysterectomy. PMID- 12746997 TI - Vaginal delivery with intrapartum pubic symphysis separation. A case report. AB - BACKGROUND: Pubic symphysis separation is an uncommon complication of pregnancy. It can occur during the antepartum, intrapartum and postpartum period. CASE: A 29 year-old woman, gravida 2, para 1, at 39 weeks' pregnancy, experienced regular labor pain and suprapubic pain for 3 hours. Her clinical presentation and physical examination led to the diagnosis of intrapartum pubic symphysis separation. Vaginal delivery was chosen because there was no cephalopelvic disproportion. The obstetric outcome was favorable, with a healthy female infant of 3,150 g. The patient underwent conservative management during the postpartum period. She was doing well at the 6-week follow-up. CONCLUSION: In the absence of obstetric indications for cesarean delivery, vaginal delivery can be achieved in cases of intrapartum pubic symphysis separation. Conservative management usually results in complete recovery. PMID- 12746996 TI - Successful treatment of recurrent, intractable hyperemesis gravidarum with methylprednisolone. A case report. AB - BACKGROUND: Severe and persistent hyperemesis gravidarum is a disabling condition. Women may request termination of pregnancy because of the intolerable symptoms and stress. CASE: A woman requested termination of pregnancy because of severe hyperemesis gravidarum. Her first three pregnancies were also complicated by severe and persistent vomiting. The vomiting was successfully treated with a short course of methylprednisolone. CONCLUSION: Methylprednisolone is an effective treatment for severe hyperemesis gravidarum and should be considered for women whose vomiting is persistent and refractory to conventional therapy. PMID- 12746998 TI - Superior vena cava syndrome in pregnancy. A case report. AB - BACKGROUND: Superior vena cava syndrome (SVCS) is an unusual condition resulting from upper body venous congestion. Little has been written about this condition and pregnancy. CASE: Despite concerns about cardiopulmonary decompensation from the hemodynamic changes of pregnancy, gestation proceeded uneventfully in a woman with SVCS. CONCLUSION: Pregnancy can be well tolerated in patients with SVCS. PMID- 12746999 TI - Fitz-Hugh-Curtis syndrome after laparoscopic tubal ligation. A case report. AB - BACKGROUND: Minimally invasive techniques are being used throughout all fields of surgery. With the increasing use and complexity of these cases, new complications will also develop. Fitz-Hugh-Curtis syndrome is an uncommon finding from the spread of infection in pelvic inflammatory disease, causing perihepatitis. CASE: A 29-year-old woman presented 2 weeks after an apparently uneventful laparoscopic tubal ligation with a complaint of right upper quadrant pain. She also had elevated liver function tests but normal ultrasound of the gallbladder. Eventually an intravenous pyelogram showed a bladder injury. Computed tomography revealed fluid in the pelvis and enhancement around the liver. During surgery, intense inflammation with multiple adhesions throughout the peritoneal cavity and around the liver were found. CONCLUSION: The findings were similar to the perihepatitis that occurs when Fitz-Hugh-Curtis syndrome complicates pelvic inflammatory disease. The unusual presentation in this patient made diagnosis very difficult and should remind physicians that unusual complications must be considered as technology evolves and spreads throughout all surgical fields. PMID- 12747000 TI - Oral contraceptives and venous thromboembolic events. PMID- 12747001 TI - The harmonic scalpel. PMID- 12747004 TI - Genetically engineered corn rootworm resistance: potential for reduction of human health effects from pesticides. AB - OBJECTIVE AND METHODS: Insecticide use, grower preferences regarding genetically engineered (GE) corn resistant to corn rootworm (CRW), and the health effects of using various CRW insecticides (organophosphates, pyrethroids, fipronil and carbamates) are reviewed for current and future farm practices. RESULTS: Pest damage to corn has been reduced only one-third by insecticide applications. Health costs from insecticide use appear significant, but costs attributable to CRW control are not quantifiable from available data. Methods reducing health related costs of insecticide-based CRW control should be evaluated. As a first step, organophosphate insecticide use has been reduced as they have high acute toxicity and risk of long-term neurological consequences. A second step is to use agents which more specifically target the CRW. CONCLUSION: Whereas current insecticides may be poisonous to many species of insects, birds, mammals and humans, a protein derived from Bacillus thurigiensis and produced in plants via genetic modification can target the specific insect of CRW (Coleoptra), sparing other insect and non-insect species from injury. PMID- 12747002 TI - Protection against hepatic ischemia-reperfusion injury in rats by oral pretreatment with quercetin. AB - OBJECTIVE: To investigate the possible protection provided by oral quercetin pretreatment against hepatic ischemia-reperfusion injury in rats. METHODS: The quercetin (0.13 mmol/kg) was orally administrated in 50 min prior to hepatic ischemia-reperfusion injury. Ascorbic acid was also similarly administered. The hepatic content of quercetin was assayed by high performance liquid chromatography (HPLC). Plasma glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase (GOT) activities and malondialdehyde (MDA) concentration were measured as markers of hepatic ischemia-reperfusion injury. Meanwhile, hepatic content of glutathione (GSH), activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and xanthine oxidase (XO), total antioxidant capacity (TAOC), contents of reactive oxygen species (ROS) and MDA, DNA fragmentation were also determined. RESULTS: Hepatic content of quercetin after intragastric administration of quercetin was increased significantly. The increases in plasma GPT, GOT activities and MDA concentration after hepatic ischemia-reperfusion injury were reduced significantly by pretreatment with quercetin. Hepatic content of GSH and activities of SOD, GSH-Px and TAOC were restored remarkably while the ROS and MDA contents were significantly diminished by quercetin pretreatment after ischemia-reperfusion injury. However, quercetin pretreatment did not reduce significantly hepatic XO activity and DNA fragmentation. Ascorbic acid pretreatment had also protective effects against hepatic ischemia-reperfusion injury by restoring hepatic content of GSH, TAOC and diminishing ROS and MDA formation and DNA fragmentation. CONCLUSION: It is indicated that quercetin can protect the liver against ischemia-reperfusion injury after oral pretreatment and the underlying mechanism is associated with improved hepatic antioxidant capacity. PMID- 12747003 TI - Calcium glucarate prevents tumor formation in mouse skin. AB - OBJECTIVE: Calcium Glucarate (Cag), Ca salt of D-glucaric acid is a naturally occurring non-toxic compound present in fruits, vegetables and seeds of some plants, and suppress tumor growth in different models. Due to lack of knowledge about its mode of action its uses are limited in cancer chemotherapy thus the objective of the study was to study the mechanism of action of Cag on mouse skin tumorigenesis. METHODS: We have estimated effect of Cag on DMBA induced mouse skin tumor development following complete carcinogenesis protocol. We measured, epidermal transglutaminase activity (TG), a marker of cell differentiation after DMBA and/or Cag treatment and [3H] thymidine incorporation into DNA as a marker for cell proliferation. RESULTS: Topical application of Cag suppressed the DMBA induced mouse skin tumor development. Topical application of Cag significantly modifies the critical events of proliferation and differentiation TG activity was found to be reduced after DMBA treatment. Reduction of the TG activity was dependent on the dose of DMBA and duration of DMBA exposure. Topical application of Cag significantly alleviated DMBA induced inhibition of TG. DMBA also caused stimulation of DNA synthesis in epidermis, which was inhibited by Cag. CONCLUSION: Cag inhibits DMBA induced mouse skin tumor development. Since stimulation of DNA synthesis reflects proliferation and induction of TG represents differentiation, the antitumorigenic effect of Cag is considered to be possibly due to stimulation of differentiation and suppression of proliferation. PMID- 12747005 TI - Improving removal efficiency of organic matters by adding phosphorus in drinking water biofiltration treatment. AB - OBJECTIVE: To investigate phosphorus limitation and its effect on the removal efficiency of organic matters in drinking water biological treatment. METHODS: Bacterial growth potential (BGP) method and a pair of parallel pilot-scale biofilters were used for the two objectives, respectively. RESULTS: The addition of phosphorus could substantially increase the BGPs of the water samples and the effect was stronger than that of the addition of carbon. When nothing was added into the influents, both CODMn removals of the parallel biofilters (BF1 and BF2) were about 15%. When phosphate was added into its influent, BF1 performed a CODMn removal, 6.02 percentage points higher than the control filter (BF2) and its effluent had a higher biological stability. When the addition dose was < 20 micrograms.L-1, no phosphorus pollution would occur and there was a good linear relationship between the microbial utilization of phosphorus and the removal efficiency of organic matters. CONCLUSIONS: Phosphorus was a limiting nutrient and its limitation was stronger than that of carbon. The addition of phosphate was a practical way to improve the removal efficiency of organic matters in drinking water biological treatment. PMID- 12747006 TI - Interaction and its solution in individual matching case-control study. AB - OBJECTIVE: To indicate the deficiency of the classical method for analyzing data on individual matching case-control study in consideration of the interaction between the study factor (exposure) and the matching factor, and to find out a proper method for handling this deficiency. METHOD: First, experimental data with 50 pairs of cases and controls were used for strata analysis according to the values of a matching factor to illustrate the possible interaction between a risk factor (exposure) and the matching factor. Second, a detailed procedure was proposed for analyzing such data. RESULTS: Interaction between the study factor and matching factor was demonstrated by using strata analysis and unconditional logistic regression analysis. Therefore the results from the classical analysis for such data might be incorrect. CONCLUSION: Data from individual matching case control study design should be dealt with strata analysis or multivariate analysis to explore and evaluate the possible interaction between the study factor and matching factor. The conclusion would be valid only after such analysis is conducted. PMID- 12747007 TI - Effect of ionizing radiation on the expression of p16, cyclinD1 and CDK4 in mouse thymocytes and splenocytes. AB - OBJECTIVE: To investigate the effect of ionizing radiation on the expression of p16, CyclinD1, and CDK4 in mouse thymocytes and splenocytes. METHODS: Fluorescent staining and flow cytometry analysis were employed for the measurement of protein expression. RESULTS: In time course experiments, it was found that the expression of p16 protein was significantly increased at 8, 24, and 48 h for thymocytes (P < 0.05, P < 0.01, and P < 0.05, respectively) and at 24 h for splenocytes (P < 0.05) after whole body irradiation (WBI) with 2.0 Gy X-rays. However, the expression of CDK4 protein was significantly decreased from 8 h to 24 h for thymocytes (P < 0.05-P < 0.01) and from 8 h to 72 h for splenocytes (P < 0.05-P < 0.01). In dose effect experiments, it was found that the expression of p16 protein in thymocytes and splenocytes was significantly increased at 24 h after WBI with 1.0, 2.0, and 4.0 Gy (P < 0.05-P < 0.01), whereas the expression of CDK4 protein was significantly decreased with 2.0 Gy for thymocytes (P < 0.05) and 0.5 6.0 Gy for splenocytes (P < 0.05-P < 0.01). Results also showed that the expression of CyclinD1 protein decreased markedly in both thymocytes and splenocytes after exposure. CONCLUSION: The results indicate that the expression of p16 protein in thymocytes and splenocytes can be induced by ionizing radiation, and the p16-CyclinD1/CDK4 pathway may play an important role for G1 arrest of thymocytes induced by X-rays. PMID- 12747009 TI - Research development of endocrine disrupting chemicals (EDCs) in water in China. AB - More and more importance has been attached to the problem of endocrine disrupting chemicals (EDCs) since 1960s. This article elaborates the recent research progress of EDCs in water and the trends in the near future in China. PMID- 12747008 TI - 3,4-methylenedioxymethamphetamine (MDMA) abuse markedly inhibits acetylcholinesterase activity and induces severe oxidative damage and liperoxidative damage. AB - OBJECTIVE: To investigate whether 3,4-methylenedioxymethamphetamine (MDMA) abuse produces another neurotoxicity which may significantly inhibit the acetylcholinesterase activity and result in severe oxidative damage and liperoxidative damage to MDMA abusers. METHODS: 120 MDMA abusers (MA) and 120 healthy volunteers (HV) were enrolled in an independent sample control design, in which the levels of lipoperoxide (LPO) in plasma and erythrocytes as well as the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and acetylcholinesterase (AChE) in erythrocytes were determined by spectrophotometric methods. RESULTS: Compared with the average values of biochemical parameters in the HV group, those of LPO in plasma and erythrocytes in the MA group were significantly increased (P < 0.0001), while those of SOD, CAT, GPX and AChE in erythrocytes in the MA group were significantly decreased (P < 0.0001). The Pearson product-moment correlation analysis between the values of AChE and biochemical parameters in 120 MDMA abusers showed that significant linear negative correlation was present between the activity of AChE and the levels of LPO in plasma and erythrocytes (P < 0.0005-0.0001), while significant linear positive correlation was observed between the activity of AchE and the activities of SOD, CAT and GPX (P < 0.0001). The reliability analysis for the above biochemical parameters reflecting oxidative and lipoperoxidative damages in MDMA abusers suggested that the reliability coefficient (alpha) was 0.8124, and that the standardized item alpha was 0.9453. CONCLUSION: The findings in the present study suggest that MDMA abuse can induce another neurotoxicity that significantly inhibits acetylcholinesterase activity and aggravates a series of free radical chain reactions and oxidative stress in the bodies of MDMA abusers, thereby resulting in severe neural, oxidative and lipoperoxidative damages in MDMA abusers. PMID- 12747010 TI - Genotoxic effects of PAH containing sludge extracts in Chinese hamster ovary cell cultures. AB - OBJECTIVE: Many studies have been conducted in order to evaluate the genotoxicity of chemicals and waste materials, which utilized in vivo test protocols. The use of animals for routine toxicity testing is now questioned by a growing segment of society. METHODS: Keeping the above fact in mind, we have conducted in the present study the genotoxicity evaluation of oily sludge samples generated from a petroleum refinery and petrochemical industry and ETP sludge from petroleum refinery using DNA damage, chromosomal aberration, p53 protein induction and apoptosis in short term in vitro mammalian Chinese Hamster Ovary cell cultures. RESULTS: It is evident from the results that the oily sludge compounds derived from petroleum refinery and petrochemical industry could cause DNA damage, chromosomal aberration, p53 protein accumulation and apoptotic cell death on exposure to oily sludge extracts in the presence of metabolic activation system (S-9 mix), however, ETP sludge extract could not cause significant genotoxicity in comparison to oily sludge extract and negative control. CONCLUSION: The effect may be attributed to polycyclic aromatic hydrocarbons present in the samples as evidenced from GC-MS. PMID- 12747011 TI - Modulation of isoflavones on bone-nodule formation in rat calvaria osteoblasts in vitro. AB - OBJECTIVE: To observe the effects of two main isoflavones, daidzein and genistein on the bone-nodule formation in rat calvaria osteoblasts in vitro. METHODS: Osteoblasts obtained from newborn Sprague-dawley rat calvaria were cultured for several generations. The second generation cells were cultured in Minimum Essential Medium supplemented with ascorbic acid and Na-beta-glycerophosphate for several days, in the presence of daidzein and genistein, with or without the estrogen receptor antagonist ICI 182780. Number of nodules was counted at the end of the incubation period (day 20) by staining with Alizarin Red S calcium stain. The release of osteocalcin, as a marker of osteoblast activity, was also determined on day 7 and day 12 during the incubation period. RESULTS: Compared with the control, the numbers of nodules were both increased by incubation with daidzein and genistein. 17 beta-estradiol was used as a positive control and proved to be a more effective inducer of the increase in bone-nodules formation that daidzein and genistein. The release of osteocalcin into culture media was also increased in the presence of daidzein and genistein, as well as 17 beta estradiol on day 7 and day 12 (day 12 were higher). The estrogen receptor antagonist ICI 182780 completely blocked the genistein- and 17 beta-estradiol induced increase of nodule numbers and osteocalcin release in osteoblasts. However, the effects induced by daidzein could not be inhibited by ICI 182780. CONCLUSION: These findings suggest that geinistein can stimulate bone-nodule formation and increase the release of osteocalcin in rat osteoblasts. The effects, like those induced by 17 beta-estradiol, are mediated by the estrogen receptor dependent pathway. Daidzein also can stimulate bone-nodule formation and increase the release of osteocalcin in rat osteoblasts, but it is not, at least not merely, mediated by the estrogen receptor dependent pathway. PMID- 12747012 TI - Effect of rat Schwann cell secretion on proliferation and differentiation of human neural stem cells. AB - OBJECTIVE: To investigate the effect of rat Schwann cell secretion on the proliferation and differentiation of human embryonic neural stem cells (NSCs). METHODS: The samples were divided into three groups. In Group One, NSCs were cultured in DMED/F12 in which Schwann cells had grown for one day. In Group Two, NSCs and Schwann cells were co-cultured. In Group Three, NSCs were cultured in DMEM/F12. The morphology of NSCs was checked and beta-tubulin, GalC, hoechst 33342 and GFAP labellings were detected. RESULTS: In Group One, all neural spheres were attached to the bottom and differentiated. The majority of them were beta-tubulin positive while a few of cells were GFAP or GalC positive. In Group Two, neural spheres remained undifferentiated and their proliferation was inhibited in places where Schwann cells were robust. In places where there were few Schwann cells, NSCs performed in a similar manner as in Group One. In Group Three, the cell growth state deteriorated day after day. On the 7th day, most NSCs died. CONCLUSION: The secretion of rat Schwann cells has a growth supportive and differentiation-inducing effect on human NSCs. PMID- 12747013 TI - Affirmative action. Psychological data and the policy debates. AB - The authors bring psychological research to bear on an examination of the policy of affirmative action. They argue that data from many studies reveal that affirmative action as a policy has more benefits than costs. Although the majority of pro-affirmative action arguments in the social sciences stress diversity, the authors' argument focuses on issues of merit. The merit-based argument, grounded in empirical studies, concludes that the policy of affirmative action conforms to the American ideal of fairness and is a necessary policy. PMID- 12747014 TI - Practicing psychology in the era of managed care. Implications for practice and training. AB - Beginning with the HMO Act of 1973, managed care, a system for controlling health care costs, rapidly expanded and gained influence as the main vehicle for health care delivery in the United States. Implementation of managed care principles in the mental health arena has generated much debate, particularly with respect to issues of quality of care. The authors briefly trace the development of managed care and evaluate its impact on the practice of psychology. The extant literature is reviewed with specific attention to issues of quality of care, confidentiality of patient information, and shifting practice patterns of clinicians. Finally, the future of professional psychology within the context of managed care is examined, and the implications of newly created mental health roles for practitioners, training programs, and organized psychology are discussed. PMID- 12747015 TI - Media ratings for violence and sex. Implications for policymakers and parents. AB - This article reviews research on the implementation of media-rating systems, parents' use and evaluation of them, and the impact of ratings on children. Although half or more of parents report using media-rating systems, understanding of various components of the systems is low, particularly for television ratings. A meta-analysis of national polls shows that parents overwhelmingly prefer that ratings specify content, rather than giving age recommendations. A second meta analysis, of experiments testing the effects of ratings on children's interest in programs, shows that ratings indicating restricted or controversial content have a deterrent effect for children under age 8 but that, by age 11 and especially for boys, the ratings show a small enticement effect. This effect occurs for both age-based and content-based ratings. Implications for policymakers and parents are discussed. PMID- 12747016 TI - Not all psychologists are classist. PMID- 12747018 TI - Comparing cultural and individual learning tendencies. PMID- 12747019 TI - The core of Confucian learning. PMID- 12747021 TI - AWP reimbursement scrutinized by several states. PMID- 12747022 TI - Non-face-to-face services deserve HMO coverage too. PMID- 12747023 TI - Compensation monitor. Economic boom of 90s left many physicians behind. PMID- 12747024 TI - Unprepared for the boomers. PMID- 12747025 TI - Despite Kaiser settlement guidelines on Web unlikely. PMID- 12747026 TI - The thin line between malpractice and benefit determination. PMID- 12747027 TI - FTC and Justice Department find some IPAs go too far in bargaining. PMID- 12747028 TI - Management of moderate to severe plaque psoriasis with biologic therapy. PMID- 12747029 TI - Promising compounds portend major management problems. PMID- 12747030 TI - Managed care outlook. Managing chronic illness becomes more crucial. PMID- 12747031 TI - Gephardt: venturing into no-man's-land. PMID- 12747032 TI - The truth about SARS. PMID- 12747033 TI - Tale of two countries. A Time investigation into what went wrong. PMID- 12747034 TI - Mother nature: political reformer. PMID- 12747035 TI - Steeped in health. Ordinary tea may help fight cancer, heart disease and, now, infection. PMID- 12747036 TI - Bernie's gift. PMID- 12747037 TI - Presidential perspectives: the ABCs of bridge building. PMID- 12747038 TI - Utilization and perceived benefits of postoperative otology in-service training for inpatient nurses. AB - This study examined the perceived effectiveness of in-service training (IST) to improve the nursing care of post-operative otology patients as a function of nursing experience. IST was administered to four distinct units in a large university hospital, examining nurses with a range of experiences in otology care. An eight-item IST evaluation instrument was developed, administered, and validated for this study. The results of this study indicate the following: a variation existed among units in the receptiveness to IST; a 15-20-minute IST session significantly improved the perceived ability to deliver quality patient care; similar effectiveness of the IST was observed for all otology care groups; and an oral IST approach was more effective than a self-study approach. Further, we found that IST increased awareness and utilization of specialty support services. PMID- 12747039 TI - Best practice forum: standard high level disinfection protocol development. AB - Gluteraldehyde-based high-level disinfection (HLD) poses significant safety risks for staff and patients as well as institutional cost concerns. A value analysis team reviewed HLD practices, available products, and environmental requirements in view of the literature and standards developed by the Occupational Safety and Health Administration (OSHA). Institutional areas were identified and standardized unit-specific educational and competency programs were established. The overall use of gluteraldehyde-based HLD was decreased in this institution based on the findings and policy and procedure modification. PMID- 12747040 TI - Highlights from the Hill: NIWI: the Nurse in Washington Internship Program--a call to all SOHN members. AB - Recent findings from the membership survey conducted in 2002 by the Society of Otorhinolaryngology and Head-Neck Nurses (SOHN) revealed that many members of SOHN were unfamiliar with the Nurse in Washington Internship program (NIWI). SOHN has been participating in the NIWI endeavor since the late eighties. To date, SOHN's NIWI interns have been either active members of the Government Relations Committee or the President-Elects of the Society. At the January 2003 Mid-Winter Board Meeting, the Board of Directors voted to expand the opportunity for participation in NIWI to SOHN's members-at-large. This article provides a brief overview of the NIWI program and describes the plan for extending this opportunity to the general membership. PMID- 12747041 TI - Of specialty interest: the Food and Drug Administration: a partner in safe practice. AB - Have you ever wondered what really goes on at the United States Food and Drug Administration (FDA)? At each meeting of the Society of Otorhinolaryngology and Head-Neck Nurses (SOHN) and the American Academy of Otolaryngology-Head and Neck Surgery (AAO-HNS) the FDA is repeatedly mentioned, and not always favorably! This article discusses the process of bringing new medical devices to market and explains how ORL (or ENT) nurses can contribute to the protection of the public health by providing medical device adverse event information to FDA. PMID- 12747042 TI - Marketing your practice. Do you need SWOT team? PMID- 12747043 TI - Make them love you... make them really, really love you. PMID- 12747044 TI - EG-ce-TRRA. How changes in tax laws affect your retirement plan. PMID- 12747045 TI - It's a long queue, too! PMID- 12747046 TI - What is amalgam? PMID- 12747047 TI - Violators? PMID- 12747048 TI - A three-sided debate on medical malpractice reform. PMID- 12747049 TI - Medicare according to Bush: don't use that "P" word. PMID- 12747050 TI - In support of patient care. An interview with Lawrence J. Majka of advocate health care. PMID- 12747051 TI - Hospitals, group purchasing organizations, and the antitrust laws. AB - Hospital members of group purchasing organizations should ensure that their GPO's business practices do not restrain trade. Hospitals should monitor their GPO to minimize their antitrust exposure. GPOs and their hospital members should assess administrative fees, bundling arrangements, duration of contracts with suppliers, and product-breakthrough policies in relation to their impact on competition and product innovation. GPOs and their hospital members should closely monitor activities of federal and state antitrust agencies in light of the agencies' renewed antitrust concerns. PMID- 12747052 TI - Merging the managed care cycle and revenue cycle to meet strategic goals. AB - By integrating the functions of the managed care cycle and the revenue cycle, providers can more readily obtain appropriate payment. The managed care cycle emphasizes strategy, while the revenue cycle emphasizes day-to-day business functions. The four phases of the managed care cycle are interlinked. Management requires information obtained from the revenue cycle to decide upon action needed in the managed care cycle. PMID- 12747053 TI - Meeting the nursing shortage head on. A round table discussion. PMID- 12747054 TI - Operational and design strategies for a reduced workforce. AB - In response to continuing workforce shortages, healthcare financial executives should encourage workflow processes and facility designs that promote staff productivity. The physical layout of care-delivery areas can affect efficiency. Facility planners should investigate opportunities to improve efficiency through consolidation of functions. Adequate staff amenities and support space can foster increased staff satisfaction and thus lead to higher retention and recruitment rates. PMID- 12747055 TI - Are you prepared for a shortage of anesthesia providers? AB - The number of anesthesia providers has not kept pace with increasing demand for their services. The supply of certified registered nurse anesthetists is decreasing. Hospital administrators need to recognize that these declining numbers will give anesthesiologists increased leverage during contract negotiations. Hospitals should prepare for negotiations with anesthesia providers by establishing process guidelines before discussions, defining minimally acceptable terms, and exploring a full range of compensation options. Hospitals that anticipate market changes and avoid hard-line responses position themselves for greatest long-term success. PMID- 12747056 TI - Transforming the revenue cycle. PMID- 12747057 TI - Political insight: contemplating Medicare reform. Interview by Eric C. Reese. PMID- 12747058 TI - Medicare: how you see it depends on where you stand. AB - Stepping outside of one's own frame of reference in regard to Medicare can yield greater understanding of the program's importance, complexities, and strengths. Hospitals in states like Pennsylvania, which has a large proportion of senior citizens, are particularly strapped by declines in payment. A national study finds Medicare beneficiaries tend to be more pleased with their level of service than are those who have private Insurance. Medicare trustees predict the program will remain solvent through 2030, although cash deficits will reemerge in 2016, creating challenges for policy-makers. PMID- 12747059 TI - You don't know what you've got till it's gone. PMID- 12747060 TI - Analyzing information technology value. PMID- 12747061 TI - Are you a center of influence? PMID- 12747062 TI - Are you tracking medication-related incidents? PMID- 12747063 TI - Designing for quality: hospitals look to the built environment to provide better patient care and outcomes. AB - With recent shifts in the marketplace, hospital boards and administrators are increasing their focus on building new facilities or renovating old ones. But rather than stick with models of the past, many of them are beginning to look at the built hospital environment as a way to promote healing, improve clinical outcomes, enhance patient safety, increase patient satisfaction, and reduce costs. At the same time, they also are finding that when changing the physical structure, they must change the internal culture of the organization as well. PMID- 12747064 TI - It's "back to the present" to find peace and contentment for you and your staff. PMID- 12747065 TI - Personal faith demonstrates healing powers. PMID- 12747066 TI - Sometimes it might just pay to say you're sorry. PMID- 12747067 TI - Ten ways to influence a leadership-challenged boss. PMID- 12747068 TI - Recruiting efforts focus on minorities, men. PMID- 12747069 TI - Testing the model for testing competency. AB - The pilot study to demonstrate the utility of the CBRDM in the practice setting was successful. Using a matrix evaluation tool based on the model's competencies, evaluators were able to observe specific performance behaviors of senior nursing students and new graduates at either the novice or competent levels. The study faced the usual perils of pilot studies, including small sample size, a limited number of items from the total CBRDM, restricted financial resources, inexperienced researchers, unexpected barriers, and untested evaluation tools. It was understood from the beginning of the study that the research would be based on a program evaluation model, analyzing both processes and outcomes. However, the meager data findings led to the desire to continue to study use of the model for practice setting job expectations, career planning for nurses, and curriculum development for educators. Although the California Strategic Planning Committee for Nursing no longer has funding, we hope that others interested in role differentiation issues will take the results of this study and test the model in other practice settings. Its ability to measure higher levels of competency as well as novice and competent should be studied, i.e., proficient, expert, and advanced practice. The CBRDM may be useful in evaluating student and nurse performance, defining role expectations, and identifying the preparation necessary for the roles. The initial findings related to the two functions as leader and teacher in the care provider and care coordinator roles led to much discussion about helping students and nurses develop competence. Additional discussion focused on the roles as they apply to settings such as critical care or primary health care. The model is useful for all of nursing as it continues to define its levels of practice and their relationship to on-the-job performance, curriculum development, and career planning. PMID- 12747070 TI - California hospitals face more disclosure legislation. PMID- 12747071 TI - A paradigm shift in cephalometric imaging. PMID- 12747072 TI - Accidental ingestion of a rapid palatal expander. PMID- 12747073 TI - Vibratory stimulation as a method of reducing pain after orthodontic appliance adjustment. PMID- 12747075 TI - Survey of orthodontic students on practice facility preferences. PMID- 12747074 TI - Surgical correction of mesially impacted mandibular second molars. PMID- 12747076 TI - Early orthopedic Class III treatment with a modified tandem appliance. PMID- 12747077 TI - PVD screenings offer revenue opportunities. PMID- 12747078 TI - HealthSouth scandal offers opportunities. PMID- 12747079 TI - Hospital-linked care continuums can create provider feedback loop. PMID- 12747080 TI - Fitch ratings forecasts negative outlook for nursing homes, CCRCs. PMID- 12747081 TI - PwC sees hospital spending jump. PMID- 12747082 TI - Solucient: study shows hospital margins rose in first half of 2002. PMID- 12747083 TI - Expert: hospital target market area must be continually re-evaluated. PMID- 12747084 TI - Hospitals face challenges of smallpox vaccination. PMID- 12747085 TI - Nutritional composition of molokhia (Corchorus olitorius) and stamnagathi (Cichorium spinosum). PMID- 12747086 TI - Nutritional composition of selected wild plants in the diet of Crete. PMID- 12747087 TI - Kanjero (Digera arvensis) and drumstick leaves (Moringa oleifera): nutrient profile and potential for human consumption. PMID- 12747088 TI - Ivy gourd (Coccinia grandis Voigt, Coccinia cordifolia, Coccinia indica) in human nutrition and traditional applications. PMID- 12747089 TI - Acerola (Malpighia glabra L., M. punicifolia L., M. emarginata D.C.): agriculture, production and nutrition. PMID- 12747090 TI - Food-based approaches to prevent and control micronutrient malnutrition: scientific evidence and policy implications. PMID- 12747091 TI - A healthy dose of privacy. A new law tries to protect patients' medical records- but has glaring gaps. PMID- 12747092 TI - 'No' in a needle. New vaccines meant to block drug highs could help break a habit or keep one from starting. PMID- 12747094 TI - [The role of nitric oxide in the pathogenesis of central nervous system damages]. AB - The review considers a role of the modulator nitric oxide--in the pathogenesis of central nervous system damages, first of all, of ischemic type. Multiple nitric oxide influences on cerebral blood flow, oxidant stress system and intracellular signaling mechanisms are shown. Investigation of nitric oxide system opens new perspectives for a therapy of acute and chronic brain ischemia. PMID- 12747093 TI - [Risk factors for subcortical arteriosclerotic encephalopathy]. AB - Subcortical arteriosclerotic encephalopathy (SAE) is a chronic progressive form of brain blood supply deficiency. Risk factors for SAE development were studied in 65 patients (42 men and 23 women, mean age 60.5 +/- 7.5 years). A control group included 31 patients (17 men and 14 women, mean age 59.3 +/- 7.4 years) with isolated clinically meaningful lacunar infarcts. A main risk factor for SAE was arterial hypertension (AH) emerging in 98.5% of the patients, which, according to twenty-four hour monitoring, differed significantly from that in the patients with isolated lacunar infarcts. In SAE, diastolic pressure was higher, systolic AP variability was detected more frequent, physiological AP decreased rarely in the nighttime, but AP fell down extremely more often. A frequency of other risk factors (ischemic disease, atrial fibrillation, diabetes mellitus, smoking, elevation of hematocrit, fibrinogen and platelet aggregation) did not differ significantly comparing to isolated lacunar infarcts. Hypercholesterolemia was detected more frequently in the controls than in the SAE patients. The study revealed that AP with hemodynamic features, pathogenetically crucial for development of disseminated arteriolosclerosis in small brain arteries and arterioles as well as for diffuse white matter damage in brain hemispheres characteristic for SAE, is a main risk factor for SAE. PMID- 12747095 TI - [Apoptosis in neuronal structures and the role of neurotrophic growth factors. Biochemical mechanisms of brain derived peptide preparations]. AB - Phenomenology of neurodegenerative disorders of any genesis corresponds to modern concepts of apoptosis as a morphobiochemical mechanism for programmed death of certain nervous cell populations. Neuroapoptosis is assumed to be a basic cause of all kind of neuropathology. Neuropeptides synthesized in certain brain regions and neurotrophic growth factors playing an important role in brain function control get involved in neurodestructive process realization as pro- or antiapoptotic components. On the basis of above concepts it is suggested that therapeutic efficacy of cerebrolysin, successfully used for therapy of wide spectrum of ischemic, neurodegenerative and other brain pathologies lies in inhibiting influence on apoptosis-dependent processes in the nervous cell. This medication including a neuropeptide and neurotrophic factors complex has many targets, which may be used for neuroapoptosis correction on different stages of pathological process. PMID- 12747096 TI - [A pathogenesis of post-stroke depression]. AB - Thirty-one patients (17 males and 14 females aged 30-74 years, mean age 61 years) with depression were examined 3-6 months after ischemic stroke. Control group included 10 sex- and age-matched healthy subjects. Neurological, psychological and psychometric tests (the Hamilton, Beck, Spilberger, mini mental scales and Quality of Life questionnaire) were conducted. A mean depression level in patients with stroke did not differ from that in the controls and did not depend on lesion size, location, neurological deficit degree. Clinically pronounced, i.e. relatively severe, post-stroke depression was detected in 22% of the patients and its severity was higher in cases of right-hemisphere stroke. Psychosocial factors were found to be the most significant in post-stroke depression. Depression occurred more often in patients, who were single, in women, in preserved intelligence and higher level of personality anxiety. Loss of job as a result of the vascular disaster proved to be not a very important psycho traumatizing factor in the group studied. PMID- 12747097 TI - [Principles of early rehabilitation of patients with stroke]. AB - Basing on general pathophysiological data, the main outlines and principles of early rehabilitation of patients with acute disturbance of cerebral blood flow are considered. Clinical ENMG analysis of peripheral neuromuscular apparatus in 32 patients with right- and left-side location of the lesion in acute and early rehabilitation periods of ischemic stroke is presented. Different variants of ENMG interrelations between paretic and intact extremities and their correlations with functional rehabilitation degree are shown. Different sanogenetic mechanisms of movement functions rehabilitation in patients with right- and left-side lesions are hypothesized. The results of early rehabilitation of patients with stroke in neurological clinic of Russian State Medical University (General Municipal Hospital N20) are presented. PMID- 12747098 TI - [Cerebral ischemic strokes in young patients with neurosyphilis]. AB - In split of a dramatic increase of syphilis incidence over the last time, neurosyphilis cases are reported relatively rare. The data on ischemic stroke of syphilis origin in 5 patients (3 males and 2 females, aged 35-43 years) are presented. A diagnosis was verified on the basis of clinical and MRT data and the results of blood and spinal fluid serological reactions. The patients had mild and moderate headache (5), movement (5) and sensitivity (3) disorders, membrane syndrome (2), hemianopsia (2), coma (1). Because of the absence of anamnesis data on syphilis survived, recognition of stroke origin was delayed and established only after positive serum and liquor tests. One patient died of multifocal brain lesion, 4 patients discharged from the hospital had differently pronounced movement disorders. PMID- 12747099 TI - [Acute disturbances of cerebral blood flow in systemic lupus erythematosus]. AB - Seventy-four patients with systemic lupus erythematosus were studied using a complex clinico-neurological and instrumental examination. Frequency of main syndromes was estimated. Characteristic features of acute cerebrovascular pathology development and risk factors for stroke in patients with systemic lupus erythematosus are presented. PMID- 12747101 TI - [Cerebrovascular reactivity in elderly patients with essential hypertension]. AB - Twenty-six patients (mean age 66.3 +/- 6.5 years) with long-term arterial hypertension were studied. In resting state the indices of cerebral hemodynamics did not differ substantially in the patients comparing to those in controls with normal arterial pressure. However, during functional tests with breath-holding and hyperventilation elderly patients with long-term mild and moderate essential arterial hypertension showed, disturbance of cerebrovascular reactivity manifesting in reduction of reactivity coefficients in response to hypo- and hypercapnic loading, earlier restoration of baseline blood flow speed in medial brain artery after hyperventilation and to breath-holding test (20%). PMID- 12747100 TI - [Disrupted circle of Willis and crisis course of arterial hypertension]. AB - Two patients, aged 17 and 55 years, with partly or completely open circle of Willis were examined using cerebral magnetic resonance angiography. A role of anatomical structure of circle of Willis and other anastomotic peculiarities of brain blood supply in cerebral vascular crises development is demonstrated. The latter can proceed as hypertensive ones on the background of emotional overloading and also reflect other features of brain blood supply in its clinical picture e.g. an absence of one of spinal arteries on the background of atherosclerosis and arterial hypertension. PMID- 12747102 TI - [The use of RR-cardiointervalometry for prognosis of disease course and outcome in patients with acute stage of ischemic stroke]. AB - The use of computed express analysis of RR-cardiointerval duration variability in patients with acute stage of ischemic stroke allowed to obtain the data on state dynamics and predict the disease outcome before the objective changes in clinical symptomatology. The recovery is observed in normalization of sigma, beta 1, beta 2, U, S-ULF, S-LF and S-HF indices. An opposite trend is observed in cases of lethal outcome. PMID- 12747103 TI - [Secondary stroke prophylaxis: the advantages of care in neurological center for specialized outpatient treatment]. AB - In neurological center for specialized course outpatient treatment, a higher level of medical care, comparing to general clinics, is provided for post- stroke patients. The continuous antiaggregants intake and modern antihypertensive therapy allowed to reduce significantly a frequency of secondary stroke in comparison to standard care for such patients in general outpatient clinics (4.6% versus 24% during a year; p < 0.05). A combined treatment in neurological center promoted a decrease of neurological deficit in 92.6% of all the cases. Positive experience of the secondary stroke prevention unit gives grounds for its introduction in clinical practice. PMID- 12747104 TI - [Analysis of brain stroke mortality in Moldova Republic]. PMID- 12747105 TI - [The experience of organization and clinical problems solution in emergency care for cerebrovascular diseases in Krasnodar]. PMID- 12747106 TI - Pros and cons of certificates. American Health Planning Association directory suggests that certificate-of-need process is regulatory in theory, not in practice. AB - As state lawmakers diluted the impact of certificate-of-need laws last year, more and more CON boards are approving a vast array of projects. Almost one-third of the 27 states that regulate acute-care hospitals gave the OK to every project they reviewed. Condell Medical Center, Libertyville, Ill., shown at left and on the cover, spent $99,300 three years ago to win state approval for its expansion project. PMID- 12747107 TI - Price check. Urged by union, CalPERS investigates alleged overpricing at Sutter's northern Calif. hospitals. PMID- 12747108 TI - Trust challenged. AHA considers involvement in charitable trust fight. PMID- 12747109 TI - Savings plan. Debate over Medicare privatization picks up steam. PMID- 12747110 TI - Two exit HCA board. Shareholder proposes pay cut for chairman, CEO. PMID- 12747111 TI - What's good for the goose.... Scrutinized Consorta gives suppliers code of conduct. PMID- 12747112 TI - Internal overhaul. Legislation proposed to reform Slidell Memorial board. PMID- 12747113 TI - Culture clash. Norton, university end plans for joint cancer facility. PMID- 12747114 TI - Not so special. Doc ownership in specialty hospitals scrutinized by healthcare panel. PMID- 12747116 TI - Looking into the future. Predictions don't always come true, but hospitals still must look ahead as aging boomers, booming technology require new solutions. PMID- 12747115 TI - A hole in the homeland defense. Fixated on smallpox, U.S. is unprepared for a more likely terror threat. PMID- 12747117 TI - By the numbers. Largest managed-care organizations. PMID- 12747118 TI - Tyranny of mind. PMID- 12747119 TI - An ocean of genes. Speed-reading all the DNA in the sea--and the gut--could reveal worlds of unknown organisms. PMID- 12747120 TI - Staying put. With CEO turnover at its lowest in years, healthcare industry insiders cite economy, board support as reasons behind the numbers. AB - Observers say the sluggish economy, combined with increased patience from hospital boards, helped push the turnover rate for hospital CEOs to an eight-year low of 14% in 2002. Thomas Dolan, left, president of the American College of Healthcare Executives, says the fallout from Sept. 11, 2001, and tough economic times may have made top executives wary of risking a job change. PMID- 12747121 TI - HHS' inspector general warns providers about joint ventures. With sham deals increasing, office cautions that suspicious cases may trigger antikickback statute. PMID- 12747122 TI - Noose tightens. Link to Scrushy alleged in criminal case. PMID- 12747123 TI - Gephardt's opening shot. More Dem hopefuls likely to offer insurance solutions. PMID- 12747124 TI - Turning volume down. Weak numbers contrast with previous predictions. PMID- 12747125 TI - Skipping the hospital. Implant campaign goes straight to the patient. PMID- 12747126 TI - Casualty of war. Hospitals jeopardized by insurance company's fallout. PMID- 12747127 TI - Notes from the lobbying front. AHA's rumored Medicaid retaliation, a ranking stir the pot in the Beltway. PMID- 12747129 TI - Lightening their load. As teaching hospitals grapple with new rules limiting residents' hours on the job, cost and management issues remain among the challenges. PMID- 12747128 TI - The docs are in charge. Physician executives branch out to lead a diverse array of institutions. PMID- 12747130 TI - Feeble giants. Despite the corporate heft of their parent companies, Amicore, Xceleron highlight the complexities of storming an entrenched info-tech market. PMID- 12747131 TI - Straight talk. New approaches in healthcare. Be aware: Medicare's Ambulatory Payment Classification poses significant risks in both lost revenue and noncompliance. PMID- 12747132 TI - Application of quality improvement theory and process in a national multicenter HIV/AIDS clinical trials network. AB - Effective clinical trials depend on the production of scientifically sound data. Clinical research coordinators monitor various activities to assure that data meet standards for timeliness and quality. Traditional methods of assuring data quality are less than optimal because they are based on correcting mistakes after they occur. Because they are focused on problem prevention, the techniques of continuous quality improvement represent a more effective means of maintaining high quality of data. This article describes the means by which principles of continuous quality improvement were incorporated into an HIV/AIDS clinical research network, as well as outcomes associated with these efforts. PMID- 12747133 TI - Quality of life in patients with pituitary tumors: a preliminary study. AB - Patients with pituitary tumors frequently complain of quality of life problems despite normal hormonal blood levels. The goals of the study were to determine whether the sickness-related quality of life reported by these patients was poorer than that expected by the general population and whether some of these complaints and areas of dysfunction were more troublesome than others. Forty three patients from a southeast Michigan pituitary disorders support group volunteered to complete a survey focusing on demographic, disease data and common complaints plus the Sickness Impact Profile (SIP). The results indicated that mental and physical fatigue were the most troublesome symptoms with sleep and libidinal complaints also of significant concern. The SIP indicated significant discontent among patients with regard to overall quality of life. Although this observational investigation seemed to underscore the fact that patients with pituitary tumors have a decreased quality of life despite apparently normal hormonal status, further subgroup analysis and subsequent surveys will be required to determine the actual significance of these findings. PMID- 12747134 TI - Costs of quality management systems in long-term care organizations: an exploration. AB - The article describes a method for measuring and reporting the costs of quality management in 11 long-term care organizations (nursing homes, home health care organizations, and homes for the elderly) and a national survey in 489 organizations providing long-term care. Site visits and a questionnaire were used to measure the existence of quality management (QM) activities and investigate the costs per QM activity in more detail. Health care organizations differentiate between regular activities and QM activities. The costs of QM activities were found to vary between 0.3% and 3.5% of the budget in three nursing homes. An extrapolation of the costs of QM activities to the entire sector shows that the long-term care sector spent between 0.8% and 3.5% of the overall budget for QM in 1999. The costs of developing and implementing QM activities are higher than the costs of monitoring. Most long-term care organizations have no insight into failure costs (i.e. the costs of quality deviations). This makes it impossible for health care organizations to draw conclusions about the cost-effectiveness of QM. PMID- 12747136 TI - The banning of Frederick Wiseman's movie, "Titicut Follies"--censoring the evidence. AB - Poor quality of care in the form of indifference and indignity can be seen as the filmmaker Frederick Wiseman shows in his classic documentary, "Titicut Follies," which was banned from public showing for 25 years. PMID- 12747135 TI - Cardiovascular point of care initiative: enhancements in clinical data management. AB - The Department of Cardiovascular Surgery at East Alabama Medical Center (EAMC) initiated a program in 1996 to improve the quality and usefulness of clinical outcomes data. After years of using a commercial vendor product and enduring a tedious collection process, the department decided to develop its own tools to support quality improvement efforts. Using a hand-held personal data assistant (PDA), the team developed tools that allowed ongoing data collection at the point of care delivery. The tools and methods facilitated the collection of real time, accurate information that allowed EAMC to participate in multiple clinical quality initiatives. The ability to conduct rapid-cycle performance improvement studies propelled EAMC's Cardiovascular Surgery Program into the Top 100 as recognized by HCIA, now Solucient, for 3 consecutive years (1999-2001). This report will describe the evolution of the data collection process as well as the quality improvements that resulted. PMID- 12747137 TI - Denials of reimbursement for hospital care. AB - Much of the negative perception of managed care focuses on fear of denials of certification for reimbursement. This study examined more than 50,000 concurrent utilization reviews completed over a four-year period (1998-2001) at a large teaching hospital. The results showed a denial rate of less than 1.5% of all patients reviewed, higher denial rates among certain clinical services, higher rates of reviews among certain services only partially explained by volume of admissions, and a lack of patient criteria to receive care in the inpatient setting as the most frequent reason given for denial. PMID- 12747139 TI - The emerging specialty pharmaceutical market segment. PMID- 12747138 TI - Outcomes can be manipulated by careful patient selection. PMID- 12747140 TI - Managing the cost drivers of medical injectables. AB - In recent years, the cost increases of medical injectables--pharmaceuticals and apparatus used to treat rare and chronic diseases--have far outstripped the rises in medical costs in general. Health plans have taken notice, and many are designing and implementing new procedures for managing these treatments. The task is complex and challenging, but identifying and addressing the cost drivers and, in particular, creating specialty pharmacy networks, hold the promise of containing the spiraling cost of medical injectables. PMID- 12747141 TI - Designing and implementing an effective specialty pharmacy service program. AB - The universe of high-tech biologic medications is expanding rapidly, and health plans are struggling to control the rising costs associated with the use of these agents. Specialty pharmaceutical networks are gaining greater visibility as a helpful tool in this effort. The author describes how to write the request for proposal and begin the decision-making and implementation processes for specialty pharmacy services. PMID- 12747142 TI - The effect of selection criteria on study results. PMID- 12747144 TI - Tough questions? Scripts provide easy answers. PMID- 12747143 TI - Disease management positively affects patient quality of life. AB - Health care costs are spiraling upward. The population of the United States is aging, and many baby boomers will develop multiple chronic health conditions. Disease management is one method for reducing costs associated with chronic health conditions. Although these programs have been proven effective in improving patient health, detailed information about their effect on patient quality of life has been scarce. This article provides preliminary evidence that disease management programs for coronary artery disease, chronic obstructive pulmonary disease, diabetes, and heart failure lead to improved quality of life, which correlates with a healthier, more satisfied, and less costly patient. PMID- 12747145 TI - Who we are. Findings from the 2002 member survey. PMID- 12747146 TI - A fresh perspective on HIM. Recent graduates embrace field's opportunities. PMID- 12747147 TI - Fighting burnout. How to identify, reduce stress for you and your employees. PMID- 12747148 TI - Breaking down barriers, boosting visibility. How to move beyond the HIM Department. PMID- 12747149 TI - A spectrum of opportunities: HIM educators needed at all levels. PMID- 12747150 TI - The dawning of a new era. AAMRL develops an international scope. PMID- 12747152 TI - Calculating costs for accounting of disclosures. PMID- 12747151 TI - Practice Brief. Ensuring legibility of patient records. PMID- 12747153 TI - Communicating change with style. PMID- 12747154 TI - Catching up with HIPAA: managing noncompliance. PMID- 12747156 TI - Understanding pelvic adhesions. How to get up to date with procedures, codes. PMID- 12747155 TI - Monitoring improper Medicare payments. New CMS programs build on OIG methods to report errors. PMID- 12747157 TI - Sepsis, related terms cause confusion for coders. PMID- 12747158 TI - SARS attacks, China shudders. PMID- 12747159 TI - Rotary vs. polio. PMID- 12747160 TI - Leadership development: perk or priority? AB - Karen Barton, Zendal Pharmaceuticals' senior vice president of human resources, was livid when COO Dave Palmer slashed her executive education budget by 75%. It must have been a mistake. Without funding there could be no in-house leadership development program, which was to be the first step toward a full-blown Zendal University. But it wasn't a mistake. Not that Palmer was against bold initiatives, but, as he patiently told Barton, sales were down 26%, and there was that $300 million debt Zendal took on when it acquired Premier Pharmaceuticals. As a result, Barton's budget wasn't the only one being cut. Palmer added that it wasn't clear what the return on investment of her proposed program--or any of her current ones for that matter--would be. Barton's analysis had been woefully short on quantitative benefits. Figuring ROI for people isn't the same as calculating the payback from a machine, Barton complained to friend and ally Carlos Freitas, head of the medical devices division. But Freitas disagreed: "If you want dollars, you have to show how you fit in with [management's] plans. You must be willing to fight for resources with the rest of us." Barton bristled: "Don't you see that my department is connected to all the others? Every division benefits from the HR budget." But she knew Freitas was right. She needed to make the case that doubling her budget was a smart move even in tough times. The question was, How? Four commentators--Susan Burnett, an HR executive at Hewlett-Packard; Mike Morrison, dean of the University of Toyota; Noel M. Tichy, professor at the University of Michigan Business School; and David Owens, vice president of Bausch & Lomb's corporate university--offer advice in this fictional case study. PMID- 12747161 TI - IT doesn't matter. AB - As information technology has grown in power and ubiquity, companies have come to view it as ever more critical to their success; their heavy spending on hardware and software clearly reflects that assumption. Chief executives routinely talk about information technology's strategic value, about how they can use IT to gain a competitive edge. But scarcity, not ubiquity, makes a business resource truly strategic--and allows companies to use it for a sustained competitive advantage. You only gain an edge over rivals by doing something that they can't. IT is the latest in a series of broadly adopted technologies--think of the railroad or the electric generator--that have reshaped industry over the past two centuries. For a brief time, as they were being built into the infrastructure of commerce, these technologies created powerful opportunities for forward-looking companies. But as their availability increased and their costs decreased, they became commodity inputs. From a strategic standpoint, they became invisible; they no longer mattered. that's exactly what's happening to IT, and the implications are profound. In this article, HBR's editor-at-large Nicholas Carr suggests that IT management should, frankly, become boring. It should focus on reducing risks, not increasing opportunities. For example, companies need to pay more attention to ensuring network and data security. Even more important, they need to manage IT costs more aggressively. IT may not help you gain a strategic advantage, but it could easily put you at a cost disadvantage. If, like many executives, you've begun to take a more defensive posture toward IT, spending more frugally and thinking more pragmatically, you're already on the right course. The challenge will be to maintain that discipline when the business cycle strengthens. PMID- 12747162 TI - Is silence killing your company? AB - Many times, often with the best of intentions, people at work decide it's more productive to remain silent about their differences than to air them. There's no time, they think, or no point in going against what the boss says. But as new research by the authors shows, silencing doesn't smooth things over or make people more productive. It merely pushes differences beneath the surface and can set in motion powerfully destructive forces. When people stay silent about important disagreements, they can begin to fill with anxiety, anger, and resentment. As long as the conflict is unresolved, their repressed feelings remain potent, making them increasingly distrustful, self-protective, and all the more fearful that if they speak up they will be embarrassed or rejected. Their sense of insecurity grows, leading to further acts of silence, more defensiveness, and more distrust, thereby setting into motion a destructive "spiral of silence." Sooner or later, they mentally opt out--sometimes merely doing what they're told but contributing nothing of their own, sometimes spreading discontent and frustration throughout the workplace that can lead them, and others, to leave without thinking it through. These vicious spirals of silence can be replaced with virtuous spirals of communication, but that requires individuals to find the courage to act differently and executives to create the conditions in which people will value the expression of differences. All too often, behind failed products, broken processes, and mistaken career decisions are people who chose to hold their tongues. Breaking the silence can bring an outpouring of fresh ideas from all levels of an organization--ideas that might just raise the organization's performance to a whole new level. PMID- 12747163 TI - Global gamesmanship. AB - Competition among multinationals these days is likely to be a three-dimensional game of global chess: The moves an organization makes in one market are designed to achieve goals in another in ways that aren't immediately apparent to its rivals. The authors--all management professors-call this approach "competing under strategic interdependence," or CSI. And where this interdependence exists, the complexity of the situation can quickly overwhelm ordinary analysis. Indeed, most business strategists are terrible at anticipating the consequences of interdependent choices, and they're even worse at using interdependency to their advantage. In this article, the authors offer a process for mapping the competitive landscape and anticipating how your company's moves in one market can influence its competitive interactions in others. They outline the six types of CSI campaigns--onslaughts, contests, guerrilla campaigns, feints, gambits, and harvesting--available to any multiproduct or multimarket corporation that wants to compete skillfully. They cite real-world examples such as the U.S. pricing battle Philip Morris waged with R.J. Reynolds--not to gain market share in the domestic cigarette market but to divert R.J. Reynolds's resources and attention from the opportunities Philip Morris was pursuing in Eastern Europe. And, using data they collected from their studies of consumer-products companies Procter & Gamble and Unilever, the authors describe how to create CSI tables and bubble charts that present a graphical look at the competitive landscape and that may uncover previously hidden opportunities. The CSI mapping process isn't just for global corporations, the authors explain. Smaller organizations that compete with a portfolio of products in just one national or regional market may find it just as useful for planning their next business moves. PMID- 12747164 TI - The high cost of accurate knowledge. AB - Many business thinkers believe it's the role of senior managers to scan the external environment to monitor contingencies and constraints, and to use that precise knowledge to modify the company's strategy and design. As these thinkers see it, managers need accurate and abundant information to carry out that role. According to that logic, it makes sense to invest heavily in systems for collecting and organizing competitive information. Another school of pundits contends that, since today's complex information often isn't precise anyway, it's not worth going overboard with such investments. In other words, it's not the accuracy and abundance of information that should matter most to top executives- rather, it's how that information is interpreted. After all, the role of senior managers isn't just to make decisions; it's to set direction and motivate others in the face of ambiguities and conflicting demands. Top executives must interpret information and communicate those interpretations--they must manage meaning more than they must manage information. So which of these competing views is the right one? Research conducted by academics Sutcliffe and Weber found that how accurate senior executives are about their competitive environments is indeed less important for strategy and corresponding organizational changes than the way in which they interpret information about their environments. Investments in shaping those interpretations, therefore, may create a more durable competitive advantage than investments in obtaining and organizing more information. And what kinds of interpretations are most closely linked with high performance? Their research suggests that high performers respond positively to opportunities, yet they aren't overconfident in their abilities to take advantage of those opportunities. PMID- 12747165 TI - Hedging customers. AB - You are a marketing director with $5 million to invest in customer acquisition and retention. Which customers do you acquire, and which do you retain? Up to a point, the choice is obvious: Keep the consistent big spenders and lose the erratic small ones. But what about the erratic big spenders and the consistent small ones? It's often unclear whether you should acquire or retain them and at what cost. Businesses have begun dealing with unpredictable customer behavior by following the practices of sophisticated investors who own portfolios comprising dozens of stocks with different, indeed divergent, histories and prospects. Each portfolio is diversified so as to produce the investor's desired returns at the particular level of uncertainty he or she can tolerate. Customers, too, are assets--risky assets. As with stocks, the cost of acquiring them is supposed to reflect the cash-flow values they are likely to generate. The authors explain how to construct a portfolio based on the notion that a customer's risk-adjusted lifetime value depends on its anticipated effect on the riskiness of the group it is joining. They also show how this approach was used to identify the best prospects for Myron Corporation, a global leader in the personalized business gift industry. The concept of risk-adjusted lifetime value has a transforming power: For companies that rely on it, product managers will be replaced by customer managers, and the current method of accounting for profit and loss- which is by product--will be replaced by one that determines each customer's P&L. Once adjusted for risk, those P&Ls will become the firm's key performance and operational metric. PMID- 12747166 TI - The nonprofit sector's $100 billion opportunity. AB - Imagine what an extra $100 billion a year could do for philanthropic and other nonprofit institutions. According to a new study, the nonprofit sector could free that amount--maybe even more--by making five changes in the way it operates. The study asked two central questions: Does the sector's money flow from its source to its ultimate use as efficiently and effectively as possible? If not, where are the big opportunities to increase social benefit? According to former senator Bill Bradley and McKinsey's Paul Jansen and Les Silverman, nonprofits could save roughly $25 billion a year by changing the way they raise funds. By distributing funds more quickly, they could put an extra $30 billion to work. Organizations could generate more than $60 billion a year by streamlining and restructuring the way in which they provide services and by reducing administrative costs. And they could free up even more money--an amount impossible to estimate--by better allocating funds among service providers. The authors admit that making those changes won't be easy. The nonprofit world, historically seen as a collection of locally focused charities, has become an enormous sector, but it lacks the managerial processes and incentives that help keep the for-profit world on track. And when the baby boomers start to retire in less than a decade, public budgets will be squeezed even more than they are today. If the nonprofit sector is to help the nation cope with the stresses ahead, it must become more efficient and challenge its traditional concepts of stewardship. PMID- 12747167 TI - Don't trust your gut. AB - Making high-stakes business decisions has always been hard. But in recent decades, it's become tougher than ever. The choices facing managers and the data requiring analysis have multiplied even as the time for analyzing them has shrunk. One simple decision-making tool, human intuition, seems to offer a reliable alternative to painstaking fact gathering and analysis. Encouraged by scientific research on intuition, top managers feel increasingly confident that, when faced with complicated choices, they can just trust their gut. The trust in intuition is understandable. But it's also dangerous. Intuition has its place in decision making--you should not ignore your instincts any more than you should ignore your conscience--but anyone who thinks that intuition is a substitute for reason is indulging in a romantic delusion. Detached from rigorous analysis, intuition is a fickle and undependable guide. And while some have argued that intuition becomes more valuable in highly complex and changeable environments, the opposite is actually true. The more options you have to evaluate, the more data you have to weigh, and the more unprecedented the challenges you face, the less you should rely on instinct and the more on reason and analysis. So how do you analyze more in less time? The answer may lie in technology. Powerful new decision-support tools can help executives quickly sort through vast numbers of alternatives and pick the best ones. When combined with the experience, insight, and analytical skills of a good management team, these tools offer companies a way to make consistently sound and rational choices even in the face of bewildering complexity--a capability that intuition will never match. PMID- 12747168 TI - Hiding the patients. PMID- 12747169 TI - The No. 1 killer of women. PMID- 12747170 TI - Unmasking a crisis. PMID- 12747171 TI - Cover yourself. It's getting easier to find affordable health insurance online. Here's how. PMID- 12747173 TI - Take a healthy interest in the risks of surgery. PMID- 12747174 TI - Should nurses practise what they preach? PMID- 12747172 TI - Weathering the storm. PMID- 12747175 TI - Value for money? PMID- 12747176 TI - Neurological observations. AB - A nursing assessment of the patient's neurological status incorporates monitoring of the patient's level of consciousness, pupil reaction and motor function, as well as observation of vital signs. The importance of all these observations, and how the nurse undertakes each stage of the process, is discussed, and the Glasgow Coma Scale is described. PMID- 12747177 TI - What you need to know about....cellulitis. PMID- 12747178 TI - Ear care. PMID- 12747179 TI - Child brain development. AB - Although the human brain is formed before birth, it continues to develop for at least 20 years. Four main developmental stages are described: before birth; birth to five years; five to 10 years; and 10 to 20 years. Specific developmental skills are attained by an individual at each of these stages. Familiarization by carers and healthcare professionals of what is normal for each stage will aid in the identification of potential problems in a child's cognitive development. PMID- 12747180 TI - A community outreach service for people with COPD. AB - This paper describes the development and evaluation of a nurse-led interagency community outreach service for patients with chronic obstructive pulmonary disease. The service improved the quality of life of the patients who accessed it. However, difficulties in maintaining the service arose from the historical structure of care delivery and policy frameworks. PMID- 12747181 TI - How I coped with....nursing and MS. Interview by David Crouch. PMID- 12747182 TI - The Anaphylaxis Campaign: Youth Workshop Programme. AB - Almost 10 million people in Britain exhibit symptoms of allergy (Royal College of Physicians and Royal College of Pathologists, 1994). A significant number have severe allergic reactions to foods and substances including: peanuts, tree nuts (almonds, walnuts, hazelnuts and brazil nuts); sesame; milk; eggs; shellfish; fish; fresh fruit; insect venom; latex; and prescribed drugs. The prevalence of peanut allergy in young children has tripled in the UK during the past decade (Grundy et al, 2002). PMID- 12747183 TI - Heart failure in primary care. AB - Heart failure is a complex syndrome that results from any structural or functional cardiac disorder that impairs the ability of the heart to function as a pump. It affects, depending on definition, between one to five per cent of the UK--half a million to three million people (Cleland et al, 2001). About 63,000 new cases are reported in the UK every year (Cowie et al, 2000). PMID- 12747184 TI - Foot assessment in GP practices for people with diabetes. AB - Diabetes increases the chance of lower-limb amputation: directly, as a result of ulcers caused by trauma and complicated by lack of sensation; and indirectly, by peripheral vascular disease. In any one patient both causes can occur. PMID- 12747185 TI - Polypharmacy and older people. AB - Polypharmacy is defined as the practice of prescribing four or more medications to the same person (Department of Health, 2001). This often occurs with older people who have concurrent disease processes, each needing a specific treatment regime (Box 1). Older people receive more prescriptions per head than any other group. The National Service Framework for Older People (DoH, 2001) shows that 5 17 per cent of hospital admissions are caused by adverse reactions to medicines. It also indicates that 6-17 per cent of older patients in hospital experience adverse drug reactions. PMID- 12747186 TI - Clinical observation on shu xin yi mai capsules for prevention of recurrent stricture after coronary artery introducing treatment. AB - Shu Xin Yi Mai Capsules ([symbol: see text]) combined with western medicine was used in the routine treatment of 22 cases who successfully received coronary artery introducing therapy (Chinese-western medicine group), and the results was compared with the 26 cases treated routinely with simple western medicine in the control group (western medicine group). It was found that both the recurrence rate of angina pectoris and the incidence rate of recurrent stricture in the Chinese-western medicine group were significantly lower than that in the control group (both P < 0.05). There was no significant difference between the two groups in expression of platelet activating molecules CD62P (alpha-granular membrane protein), CD63 (lysosome intact membrane protein) and CD41 (glucoprotein IIb) before the treatment, but with a significant difference after the treatment (P < 0.05). PMID- 12747187 TI - The combined use of tonification and purgation in treatment of fatty liver. PMID- 12747188 TI - TCM treatment of renal amyloidosis--a report of one case. PMID- 12747189 TI - Prof. Cao Hongxin's experience in treating viral myocarditis with xuefu zhuyu tang. PMID- 12747190 TI - Treatment of interstitial peumopathy by fei tong oral liquid in the malignant tumor patients after radio- and/or chemotherapy. AB - Fei Tong Kou Fu Ye ([symbol: see text]) Fei Tong Oral Liquid) was used to treat 30 cases of interstitial pneumopathy after radio- and/or chemotherapy. In comparison with the control group (15 cases) treated with hormones, the therapeutic effects in improving dyspnea, cough, respiratory rate, cyanosis, findings in X-films and CT examination, partial pressure of oxygen in artery, FVC and VC were found significantly better (P < 0.05). The total effective rate obtained was 83.33%. PMID- 12747191 TI - Clinical observation on treatment of rheumatoid arthritis with fengshi xiandan in 53 cases. AB - We have successfully developed Fengshi Xiandan ([symbol: see text]) and used it to treat 53 cases of rheumatoid arthritis (RA), with indomethacin used to treat another 53 cases of RA as the controls. The results showed that the rate of cure and obvious effect was 54.7% and the total effective rate was 92.4% in the treatment group, as compared to 26.4% and 66.0% respectively in the control group, with an obvious difference in curative effect between the two groups (P < 0.01). The improvement of all the main clinical symptoms and signs except arthralgia in the treatment group was superior to that in the control group (P < 0.05 or P < 0.01). The improvement of erythrocyte sedimentation rate (ESR), rheumatoid factor (RF), C-reaction protein (CRP), immunoglobulin (Ig), T lymphocyte subgroups, micro-circulation in nail fold, and red blood cell (RBC) and hemoglobin (Hb) of anemia patients in the treatment group was superior to that in the control group (P < 0.05 or P < 0.01). There was an obvious difference (P < 0.05 or P < 0.01) in superoxide dismutase (SOD) of RBC and lipid peroxide (LPO) of plasma before treatment in the two groups as compared to normal group (P < 0.05 or P < 0.01). The level of SOD and LPO after treatment in the treatment group obviously tended to normal as compared to that before treatment (P < 0.05 or P < 0.01). The reduction of LPO in the treatment group was obviously superior to that in the control group (P < 0.01). The result shows that Fengshi Xiandan ([symbol: see text]) with an obvious curative effect on RA can not only control the symptoms of RA but also improve its pathologic damage and prevent its development. PMID- 12747192 TI - Kang wei granules in treatment of gastropathy related to Helicobacter pylori infection. AB - Kang Wei Granules, a granular preparation for strengthening the spleen and replenishing Qi and for clearing away heat and resolving dampness, was used in the treatment of 288 cases of gastropathy related to Helicobacter pylori infection. The effects were compared with De Nol triple therapy in the control group of 74 cases. The therapeutic results showed that Kang Wei Granules was superior to the western drugs in improving the principal symptoms of deficiency of the spleen and stomach, and retention of damp-heat in the interior (P < 0.05). PMID- 12747193 TI - The therapeutic effects of the radiotherapy plus TCM treatment observed in senile non-parvicellular lung cancer patients at the late stage. AB - 47 senile non-parvicellular lung cancer patients at stage III or IV were randomly divided into a treatment group (26 cases) treated by radiotherapy plus traditional Chinese medicine (TCM) and a control group (21 cases) treated only by radiotherapy for observation of the therapeutic effects. The patients in the treatment group orally took Chinese medicine during and after the radiotherapy. There was no obvious difference in short-term therapeutic effects between the two groups, but the long-term curative effects in the treatment group was obviously superior to that in the control group (P < 0.05 or P < 0.01). CONCLUSION: Radiotherapy plus TCM can prolong the survival period for senile non parvicellular lung cancer patients. PMID- 12747194 TI - TCM treatment of extrasystole with huanglian shengmai yin--a report of 357 cases. AB - The previous experimental studies have demonstrated that addition of Huang Qi ([symbol: see text] Radix Astragali) to the formulated recipe Sheng Mai Yin ([symbol: see text] Decoction for Pulse-activation) exerts the effects of strengthening the myodynamia, increasing the coronary flow, improving myocardial metabolism, and resisting the arrhythmia. The active component of Huang Lian ([symbol: see text] Rhizoma Coptidis) can prolong the myocardial action potential and antagonize the chloroform-, aconitine-, barium chloride-, epinephrine- or coronary ligation-induced arrhythmia by blocking the calcium channel. Ku Shen ([symbol: see text] Radix Sophorae Flavescentis) contains matrine and flavones, which act as quinidine to decrease the excitability of the myocardium, prolong the refractory period, and inhibit the ectopic cardiac rhythm. And Dan Shen ([symbol: see text] Radix Salviae Miltiorrhizae) has the action of improving the ischemic state of the myocardium by dilating the coronary vessels. In conclusion, the definite therapeutic effects of Huang Lian Sheng Mai Yin in treating ventricular, atrial and nodal arrhythmia suggests that the prescription is rational and accords with the therapeutic principle of TCM. Except discomfort in the gastric cavity and poor appetite experienced by some patients, there is no toxic or adverse reaction. PMID- 12747195 TI - Clinical observation on acupunture treatment of piriformis syndrome. PMID- 12747196 TI - Clinical experience in electro-acupuncture treatment. PMID- 12747197 TI - Acupuncture for treating postoperative enteroparalysis. PMID- 12747198 TI - Protrusion of lumbar intervertebral disc treated by ultra short wave and the point injection in 40 cases. PMID- 12747200 TI - Acupuncture treatment of facial paralysis caused by craniocerebral trauma in 50 cases. PMID- 12747199 TI - Clinical application of the jiuwei point. PMID- 12747201 TI - The effects of acupuncture on blood pressure in different patients. AB - Eight-seven patients with different types of diseases were treated with acupuncture, and the changes of their blood pressure were investigated. The results showed that the total level of systolic pressure in the 87 cases decreased (P < 0.05). Especially, the systolic pressure in patients accompanied with hypertension decreased significantly. It is suggested that when acupuncture is adopted to treat other diseases, it can decrease the systolic pressure to a certain extent in patients accompanied with hypertension, but it does not exert significant influence on both systolic and diastolic pressure in patients with normal blood pressure. PMID- 12747202 TI - Treatment of 50 cases of sciatica by needling zanzhu and fengchi. PMID- 12747203 TI - Electroacupuncture at ciliao and huiyang for treating neuropathic incontinence of defecation and urination in 30 cases. PMID- 12747204 TI - Acupuncture treatment for 157 cases of anxiety neurosis. PMID- 12747205 TI - Treatment of phlegm- and heat-induced insomnia by acupuncture in 120 cases. PMID- 12747206 TI - Zusanli point injection for treating leukopenia induced by radio-chemotherapy. PMID- 12747207 TI - Preliminary investigation on regulating effects of different TCM treatments on transcription of the correlated genes of liver cancer in rats. AB - The regulating effects of TCM treatments including clearing away heat and toxic materials, promoting blood circulation and removing blood stasis, and strengthening the spleen and regulating qi on the oncogene transcription were observed in the liver cancer model rats. The preliminary results indicated that the mRNA levels of H-ras N-ras and K-ras, and signal molecules correlated with the ras/MAPK signal transduction pathway were down-regulated by the different TCM treatments in varying degrees. Also, the regulating effects of the treatments on differently-displayed genes were discrepant. It is suggested that the molecular mechanisms of the TCM treatments for liver cancer was complex with different target genes. PMID- 12747208 TI - The present situation of TCM treatment for diabetes and its researches. PMID- 12747210 TI - Why are fruits and vegetables so healthy? PMID- 12747209 TI - Acupuncture treatment of melancholia. PMID- 12747211 TI - Special characteristics of non-nutrient food constituents of plants- phytochemicals. Introductory lecture. PMID- 12747212 TI - Vegetable and fruit: the evidence in their favour and the public health perspective. AB - There is strong evidence that the intake of vegetables and fruits reduces the risk of cardiovascular disease (CVD) and is inversely associated with several forms of cancer. In contrast, information concerning specific macro- or micronutrients in relation to chronic diseases is limited and largely inconclusive. The beneficial role of vegetable and fruit consumption can also be inferred by considering the health effects of two dietary patterns, the Mediterranean and Japanese ones, in both of which the consumption of plant foods holds a prominent position. Time-trend data, retrieved from the DAFNE databank on the vegetable and fruit availability in four European countries (Greece, Ireland, Italy and Norway) indicate that, during the last decade, fruit availability decreased in Greece and Italy and increased in Ireland and Norway, whereas vegetable availability decreased only in Italy. In Greece, Italy and Norway, the daily fruit availability was higher than that of vegetables, a dietary pattern not in accordance to recommendations for higher vegetable consumption. This information, which is crucial for nutrition policies and health education, also demonstrates the value of the DAFNE surveillance system. PMID- 12747213 TI - Plant foods versus compounds in carcinogenesis; observational versus experimental human studies. AB - The protective role of plant foods and its constituents in cancer prevention is under renewed debate since the results of recent observational studies on colorectal cancer as well as large-scale human experimental studies on colorectal adenoma recurrence are disappointing. However, most short-term experimental human studies do show that plant foods favourably modulate potential cancer-preventive mechanisms. Which methodological pitfalls may explain the inconsistencies within and between different study designs? What are the advantages and limitations of the different study approaches? Observational studies do have the advantage to study the population at large with ultimate disease as the study endpoint. These studies are limited by the difficulty to estimate intake of individual compounds by questionnaires and the lack of biological markers of relevant exposure. Controlled experimental short-term studies in humans rely on biological markers of disease as intermediate endpoints. Relatively low sensitivity and specificity of these markers may complicate extrapolation of results. In the case of long term and large-scale human intervention studies with disease endpoints, issues such as time, dose and duration of intervention, compliance and choice of the study population influence the interpretation of results. An integrated approach combining designs, and implementing new techniques to identify biomarkers, may clarify the role of plant foods in carcinogenesis. PMID- 12747214 TI - Transformation of flavonoids by intestinal microorganisms. AB - Fruit, vegetables and cereals contain a wealth of secondary plan metabolites which have been implicated in the promotion of health. To understand the mechanism of their action it is necessary to gain more information on their fate in the body following ingestion. A certain proportion of ingested secondary plant constituents may escape absorption in the small intestine and therefore undergo transformation by intestinal microorganisms or enterohepatic circulation. To study the transformation of secondary plant metabolites by bacteria, Eubacterium ramulus was isolated from human feces and incubated with selected flavonoids. E. ramulus is a strictly anaerobic bacterium which was found to be present in the gastrointestinal tract of most individuals investigated. E. ramulus cleaves the ring system of several flavonols and flavones giving rise to the corresponding hydroxyphenylacetic and hydroxyphenylpropionic acids, respectively, as well as acetate and butyrate. Degradation pathways were proposed based on the intermediates detected by high performance liquid chromatography (HPLC) and HPLC coupled with mass spectrometry (LC-MS) and the detection of enzymes that catalyze reactions such as taxifolin isomerization, phloretin hydrolysis and phloroglucinol reduction. The dearomatizing phloroglucinol reductase, presumably part of all flavonoid degradation pathways, was purified and characterized. The gene encoding phloretin hydrolase was cloned from a E. ramulus gene library taking advantage of a newly developed fluorescence test for activity screening. Moreover, a new intermediate was discovered and identified by MS and 1H and 13C NMR analysis as alphitonin. To investigate the degradational potential of E. ramulus under in vivo conditions, germfree rats were associated with E. ramulus. Following the intragastric application of quercetin-3-glucoside, urine and feces of gnotobiotic rats were analyzed for degradational products originating from quercetin-3-glucoside. In feces of rats monoassociated with E. ramulus, 3,4 dihydroxyphenylacetic acid was found, indicating that this organism is able to cleave quercetin under in vivo conditions. To investigate in which way the dietary flavonoid content affects the cell counts of E. ramulus in the human intestinal tract, twelve human subjects consumed a flavonoid-free diet for one week and at one point during this period a large dose of flavonoids. Fecal samples from both phases of the study were analyzed by in-situ hybridization for total bacterial counts and counts of E. ramulus. Total cell counts and the cell counts of E. ramulus decreased significantly during the flavonoid-free period, while there was an increase in the E. ramulus counts of up to 10-fold during the flavonoid-rich period indicating that dietary secondary plant metabolites may have an influence on the intestinal microflora. E. ramulus is also capable of converting the isoflavonoids genistein and daidzein to the products 2-(4 hydroxyphenyl)-propionic acid and O-desmethylangolensin, respectively. PMID- 12747215 TI - Non-nutritive bioactive food constituents of plants: tocopherols (vitamin E). AB - An increasing body of data, especially the in vitro and cell culture studies support protecting effects of tocopherols. They indicate that there is a selective tocopherol transport in the human liver with a higher affinity for alpha-tocopherol, however, they also show that in food, gamma- and delta tocopherol are a more potent antioxidant than alpha-tocopherol. Tocopherols as a group are the key antioxidants in human cell membranes and are also important in protecting the LDL particles. These LDL particles and other similar intermediate markers of oxidative stress show a good response to tocopherol intakes. Published data underline the role of tocopherols as protecting agents against oxidative stress. They therefore allow the assumption that alpha-tocopherol is probably effective in preventing atherosclerosis, although no proven direct relation to the outcome of the disease itself exists. PMID- 12747216 TI - Non-nutritive bioactive constituents of plants: lycopene, lutein and zeaxanthin. AB - Lycopene, lutein, and zeaxanthin are major carotenoids in human blood and tissues but unlike beta-carotene do not contribute to vitamin A supply. These carotenoids are efficient antioxidants quenching singlet molecular oxygen which is formed in photooxidative processes and thus may contribute to the prevention of light exposed tissue, skin and eyes, from light-induced damage. Increasing lycopene intake by daily consumption of tomato paste over a period of ten weeks provides protection against erythema formation following UV-irradiation. Lycopene and other carotenoids may be used as oral sun protectants and contribute to the maintenance of skin health. The yellow color of the macula lutea is due to the presence of the carotenoid pigments lutein and zeaxanthin. These macula carotenoids are suggested to play a role in protection against light-dependent damage. Filtering of blue light and scavenging of reactive intermediates generated in photooxidation are considered to be the underlying protective mechanisms. Epidemiological studies provide evidence that an increased consumption of lutein is associated with a lowered risk for age-related macular degeneration, a disease with increasing incidence in the elderly. PMID- 12747217 TI - Non-nutritive bioactive food constituents of plants: bioavailability of flavonoids. AB - Flavonoids are polyphenols widely distributed in the plant kingdom, and are present in fruits and vegetables regularly consumed by humans. In vitro metabolic studies of flavonoids in rat liver microsomes identified the 3',4'-dihydroxylated derivatives as the major metabolic endpoint. However, in vivo in rats almost none of this metabolite and only minor amounts of the 4'-monohydroxylated derivative was produced. Flavonoids with the 4'-monohydroxylated structure were generally not metabolised and were excreted unchanged in urine in higher amounts than other flavonoids investigated. It has for long been a controversy, whether flavonoids are absorbed as the intact glycoside or whether they have to be hydrolysed to the free aglycon prior to absorption. Recent data suggest that beta-glucosidases and maybe also lactase phlorizin hydrolase (LPH) in the small intestine are capable of hydrolysing flavonoid glucosides and these compounds are thus taken up as the free aglycon and not as the intact glycosides. LC-MS analyses of 12 dietary flavonoids in human urine showed that no flavonoid glycosides were excreted, and that the citrus flavanones and phloretin are excreted in higher amounts than the flavonols. Furthermore, total flavonoid excretion may be a useful biomarker for habitual fruit and vegetable consumption. PMID- 12747218 TI - Antioxidant actions of polyphenols in humans. AB - Many polyphenols are potent antioxidants in foods and model systems and they have therefore very naturally been linked with the hypothesis that their redox actities may confer them with specific health benefits. Their prevalence in plant derived foods, which are generally accepted as healthy has supported this view and inspired researchers to conduct human intervention trails with polyphenol rich food items in order to investigate their ability to counteract oxidative stress. Several biomarkers have gained widespread use to assess oxidative damage and antioxidative defence capabilities in humans. These markers pioneer our knowledge about factors related to oxidative stress in proteins, lipids and DNA and present results indicate that oxidative damage may be very localised and that refined markers may be necessary in order to disentangle the complex local factors which determine the extent of oxidative damage in different molecular structures. The present text reviews the human short-term intervention studies with polyphenol-rich foods, which address their impact on biomarkers of oxidative damage and antioxidative defence. None of the oxidative damage markers seem to be consistently affected by polyphenol-rich foods or to be consistently related to one another. The most consistent finding regarding antioxidative defence markers is a postprandial effect on plasma antioxidative capacity after ingestion of foods rich in catechins and complex procyanidins. PMID- 12747219 TI - Potential risks and benefits of phytoestrogen-rich diets. AB - Interest in the physiological role of bioactive compounds present in plants has increased dramatically over the last decade. Of particular interest in relation to human health are the class of compounds known as the phytoestrogens, which embody several groups of non-steroidal oestrogens including isoflavones & lignans that are widely distributed within the plant kingdom. Data from animal and in vitro studies provide plausible mechanisms to explain how phytoestrogens may influence hormone dependent states, but although the clinical application of diets rich in these oestrogen mimics is in its infancy, data from preliminary studies suggest potential beneficial effects of importance to health. Phytoestrogens are strikingly similar in chemical structure to the mammalian oestrogen, oestradiol, and bind to oestrogen receptors (ER) with a preference for the more recently described ER beta. This suggests that these compounds may exert tissue specific effects. Numerous other biological effects independent of the ER (e.g. antioxidant capacity, antiproliferative and antiangiogenic effects) have been ascribed to these compounds. Whether phytoestrogens have any biological activity in humans, either hormonal or non hormonal is a contentious issue and there is currently a paucity of data on human exposure. Much of the available data on the absorption and metabolism of dietary phytoestrogens is of a qualitative nature; it is known that dietary phytoestrogens are metabolised by intestinal bacteria, absorbed, conjugated in the liver, circulated in plasma and excreted in urine. Recent studies have addressed quantitatively what happens to isoflavones following ingestion--with pure compound and stable isotope data to compliment recent pharmacokinetic data for soy foods. The limited studies conducted so far in humans clearly confirm that soya isoflavones can exert hormonal effects. These effects may be of benefit in the prevention of many of the common diseases observed in Western populations (such as breast cancer, prostate cancer, menopausal symptoms, osteoporosis) where the diet is typically devoid of these biologically active naturally occurring compounds. However since biological effects are dependent on many factors including dose, duration of use, protein binding affinity, individual metabolism and intrinsic oestrogenic state, further clinical studies are necessary to determine the potential health effects of these compounds in specific population groups. However we currently know little about age related differences in exposure to these compounds and there are few guidelines on optimal dose for specific health outcomes. PMID- 12747220 TI - Non-nutritive bioactive constituents of plants: phytosterols. AB - Normal human diet contains small amounts of phytosterols, mainly sitosterol and campesterol. Intestinal absorption of these plant sterols is low, about one tenth of that of cholesterol, such that their serum concentrations are also low, about 0.1 to 1% of the cholesterol levels. Like cholesterol they are transported by lipoproteins, mainly by LDL, and secreted unchanged in bile. Addition of plant sterols, or especially of their delta-5 saturated derivatives plant stanols into diet as fat-soluble esters inhibit cholesterol absorption and lower serum cholesterol similarly in short-term studies. Long-term consumption of plant stanol esters lowers serum cholesterol to the extent expected to reduce clinical manifestation of coronary heart disease by over 20% without detectable side effects, cholesterol lowering being especially effective in combination with cholesterol synthesis inhibitors statins. PMID- 12747221 TI - Non-nutritive bioactive constituents of plants: dietary sources and health benefits of glucosinolates. AB - A high intake of cruciferous vegetables is associated with a reduced risk of cancer, particularly lung and those of the gastrointestinal tract. This protective effect has been linked to the presence of glucosinolates in these vegetables. Certain metabolites of the glucosinolates, particularly the isothiocyanates and nitriles have been shown to both modify xenobiotic metabolising enzymes and induce cell cycle arrest and apoptosis. Different metabolites are more or less effective in mediating each response depending on the model system employed. It is likely that a combination of these responses explains the chemo-preventive characteristics of Brassicas and that a combination of different cruciferous vegetables will provide optimal protection. PMID- 12747222 TI - Dependence of tocopherol stability on different cooking procedures of food. AB - The influence of the kind of treatment during the preparation of hot meals (roasting, baking, stewing) on the stability of the tocopherols, which were added to the food through the margarines used, were examined in different food matrices. All preparation methods resulted in a loss of tocopherols. They could be divided into a group of only small loss of tocopherols (lower than 20%) and into a second group with nearly 70% loss of tocopherols (roasting of meat, steaming of peas). In the first group losses of tocopherols were directly associated with the fatty acid patterns of the used margarines, especially with the share of polyunsaturated fatty acids. On the other hand, during the roasting of meat nearly 70% of the tocopherols were destroyed by direct oxidation without a prohibitive effect on the lipid oxidation. The extreme high loss of tocopherols during the steaming of peas may be caused by the catalytic effect of the metal surface of the used pot. There were no losses of tocopherols observed when the experiments were repeated under identical conditions in an inert glass pot. Only tocopherol losses during baking may have a nutritional importance because of the great fat amount which are taken in with cookies. PMID- 12747223 TI - Influence of cooking process on phenolic marker compounds of vegetables. AB - Phenolic compounds are secondary plant metabolites which have long been associated with flavor and color characteristics of fruits and vegetables. These phenolic compounds attract great interest due to their postulated health protecting properties. However, adequate intakes and absorption rate of phenolic compounds are necessary for these beneficial effects. Until now, little is known about alterations of phenolic compounds content by the cooking process. In the present study, the influence of different volumes of cooking water on the amount of selected phenolic marker compounds resting in the vegetables was assessed. In zucchini, rutin was quantified as a marker for flavonoid glycosides. Chlorogenic acid, representative of phenolic acids was analyzed in carrots. In beans, rutin and quercitrin, both belonging to flavonoid glycosides, were investigated. In potatoes, chlorogenic and caffeic acid were determined. The cooking of zucchini, beans and carrots with smaller amounts of water resulted in significant higher content of phenolic phytochemicals in the vegetables compared to cooking with larger water volumes. For potatoes, which showed great variations in content of phenolic acids after cooking, no significant differences in phenolic acids was observed. It can be concluded from these observations, that real intakes of phenolic compounds from cooked vegetables are lower and that the amounts consumed are therefore overestimated. PMID- 12747224 TI - Mode of conception, placental morphology and perinatal outcome of twin gestations. AB - OBJECTIVE: To compare the perinatal outcome and placental morphology of twins conceived by assisted reproductive technologies (ART) or natural conception (NC). METHODS: The present retrospective study included 88 twin pairs. Methods of ART included in vitro fertilization, intrauterine insemination, embryo transfer and induced ovulation. Placental morphology was described by chorionicity, amnionicity, placental weight, umbilical cord insertion and the number of cord vessels. Perinatal outcomes included maternal age, gestational age, birth weight (BW), BW discordance and mode of delivery. RESULTS: The mothers of ART twins were 4 years older than NC mothers. ART twins delivered 2.2 weeks earlier than NC twins. The BW of ART twins A and B were less than NC twins A and B. There was no difference in the proportion of discordant twins in either group. Dichorionic diamniotic (DD) placentas accounted for the majority of placentas in both groups. There were no monochorionic-monoamniotic placentas in ART pairs, but there were 2.9% in NC pairs. Mode of conception had no effect on placental weight in DD or monochorionic-diamniotic twins in either group except for larger DD fused placentas in the ART group. CONCLUSION: ART twin pairs were delivered at an earlier gestational age at lower birth weights to older mothers predominantly by cesarean section. This perinatal outcome was not reflected by placental morphology. PMID- 12747225 TI - The effects of birth weight discordance and prematurity on twin neonatal outcome. AB - OBJECTIVE: The purpose of this study was to determine the effect of birth weight discordance (BWD) and prematurity on twin neonatal outcome by measuring Apgar scores and blood glucose concentrations at birth. METHODS: This was a retrospective cross-sectional study. All twins born at NY Methodist Hospital between January 1998 and December 2001 were eligible. Excluded were fetal deaths, maternal systemic disease, maternal infections, major neonatal malformations, neonates with birth weight < 500 g and multiple gestation greater than 2. BWD was defined as intrapair difference in birth weight expressed as percentage of the larger twin greater than 20% and prematurity as gestation of less than 37 weeks. Hypoglycemia was defined as blood glucose concentration < 40 mg/dl. RESULTS: There were 75 (48%) preterm and 80 (52%) term twin pairs. Nineteen (12%) of the preterm and 15 (9.6%) of the term showed BWD. Mean Apgar scores at 1 and 5 minutes were higher for the term twins as expected. Neither prematurity nor hypoglycemia was present predominantly in BWD twins. Twelve of the preterm and six of the term BWD twins showed evidence of hypoglycemia. CONCLUSION: Hypoglycemia in the early hours of postnatal life is relatively common in preterm BWD twins. Blood glucose measurement using reagent strips is an efficient way of monitoring blood glucose concentrations in these infants. PMID- 12747226 TI - Esophageal atresia and tracheoesophageal fistula: the impact of prenatal suspicion on neonatal outcome in a tertiary care center. AB - AIMS: To determine the impact of antenatal suspicion of esophageal atresia (EA) with or without tracheoesophageal fistula (TEF) on neonatal outcome. METHODS: Retrospective review of all neonates with EA who received prenatal care including fetal ultrasound and delivery at our institution from 1990-2001. Cases with suspected EA on prenatal ultrasound (hydramnios and/or an absent stomach bubble) were identified. Neonatal outcome variables for the group suspected antenatally and the group diagnosed postnatally were compared. Mann Whitney U and Fischer exact tests were used in analysis. RESULTS: Twenty-two patients met inclusion criteria. Nine cases (40.9%) had prenatal ultrasound findings associated with EA/TEF. There was no statistically significant difference in the incidence of preterm delivery, intrauterine growth restriction, respiratory distress syndrome, additional anomalies or neonatal death, birth weight, requirement for preoperative and postoperative mechanical ventilation or length of hospital stay between the prenatally suspected and postnatally diagnosed groups. There were two neonatal demises: one had trisomy 18 and one was born prematurely at 29 weeks. CONCLUSIONS: In our experience, prenatal detection of ultrasound findings associated with EA/TEF does not affect neonatal outcome or identify a group at increased risk for neonatal morbidity and mortality. Our favorable outcomes, with or without prenatal suspicion, may reflect the comprehensive care readily available at a tertiary care facility. Larger series need to be studied to exclude the possibility of a type II error. PMID- 12747227 TI - Microbial invasion of the amniotic cavity at birth is associated with adverse short-term outcome of preterm infants. AB - AIMS: To determine the frequency and clinical significance of microbial invasion of the amniotic cavity at the time of delivery in preterm infants. METHODS: Prospective cohort study during June 2001 and January 2002. Preterm infants < 33 + 6 weeks of gestation who had amniotic fluid and placental tissue sampled for culture during cesarean section were included. RESULTS: Of a total of 80 neonates, 42 had negative culture results, 22 had growth of Ureaplasma urealyticum, and 16 had growth of other pathogens. Isolation of Ureaplasma urealyticum was associated with a decreased risk of developing hyaline membrane disease after birth but a more than 20 times increased risk of developing chronic lung disease. Patients with growth of other pathogens had a significantly higher mortality than patients with negative culture results. CONCLUSIONS: Isolation of miroorganisms from the amniotic cavity at birth is associated with an adverse outcome of the preterm infant. In the light of extremely small numbers of positive blood cultures in preterm infants after birth, we consider it reasonable to recommend routine culturing of amniotic cavity tissues/fluid obtained during cesarean section in order to increase the identification rate of pathogens potentially involved in the pathogenesis of perinatal infections. PMID- 12747228 TI - Is treatment with vaginal pessaries an option in patients with a sonographically detected short cervix? AB - OBJECTIVE: The purpose was to determine the effect of vaginal pessaries in patients at risk for spontaneous preterm birth (SPB). STUDY DESIGN: Transvaginal sonography (TVS) was longitudinally performed to measure cervical length (CL) in 258 singleton at risk for SPB and 282 twin pregnancies. Pairs with or without treatment were matched for gestational age and the CL at examination. RESULTS: In 4 singleton and 7 twin pregnancies the CL was < 15 mm before 24 weeks, the mean interval between pessary insertion and delivery was 13 + 2 and 12 + 5 weeks respectively. For the matched control analysis, 12 pairs with singleton and 23 pairs with twin pregnancies were compared. For singleton pregnancies, the mean interval between TVS and delivery was 99 (70-134) days in the treatment and 67 (2 130) days in the control group (p = 0.0184), the mean gestational age at delivery was 38 (36 + 6-41) and 33 + 4 (26-38) weeks respectively (p = 0.02). For twin pregnancies, the interval was 85 (43-129) days in the treatment and 67 (21-100) days in the control group (p = 0.001), gestational age at delivery was 35 + 6 (33 37 + 4) and 33 + 2 (24 + 4-37 + 2) respectively (p = 0.02). Within singleton pregnancies with pessary, there was no SPB < 36 weeks compared to 6/12 cases in the control group (p < 0.001). Within twin pregnancies, the rates were 8/23 cases with SPB < 36 weeks but none < 32 weeks, compared to 12/23 cases with SPB < 36 weeks and 7/23 cases < 32 weeks in the control group (p < 0.001). CONCLUSIONS: Insertion of a vaginal pessary may be a cost-effective preventive treatment in patients at risk for SPB. Prospective controlled trials are needed. PMID- 12747229 TI - The effects of intramuscular administration of corticosteroids on the induction of parturition. AB - Our purpose was to examine the hypothesis that corticosteroids, when administered intramuscularly, can enhance the labor process and reduce the time interval between the induction and the active phase. A randomized, controlled study was conducted on 66 women with gestational age of 41 weeks and over and favorable cervix (bishop score > or = 7). The study group (n = 32) received 10 mg of dexamethasone phosphate intramuscularly in two doses at an interval of 12 hours, and the day after the enrolling administration intravenous oxytocin was given. The control group (n = 33) received only intravenous oxytocin 24 hours after enrolling. The number of patients to enter the active phase of labor was significantly higher in the study group than in the control group (n = 33 [100%] vs n = 29 [87.9%], p < 0.039). The mean time interval between induction of labor and the active phase was significantly shorter in the study group than in the control group (1.7 +/- 1.5 hours vs 4 +/- 1.7, P < 0.0001), and the mean of oxytocin dose was significantly lower in the study group (1.15 +/- 1.5 u vs 4.16 +/- 2.5 u, P < 0.0001). Induction of labor with the use of intramuscularly injected dexamethasone phosphate reduced the time interval between the induction of labor and the active phase. PMID- 12747230 TI - Perinatal significance of diagnosing glucose intolerance during pregnancy with portable glucose meter. AB - OBJECTIVE: To evaluate the effect on perinatal outcome of replacing glucose measurements for the diagnosis of gestational diabetes mellitus in central laboratories with those obtained by portable glucose meter. STUDY DESIGN: Simultaneously, venous blood samples were taken at the 100-g glucose tolerance test, and finger capillary blood glucose levels were also measured with a portable glucose meter in 180 pregnant patients. Patient management was according to the venous plasma glucose results. Following deliveries, the women were reallocated to diagnostic groups by both methods of measurements and by using the same cutoff values, and the outcome data were compared. RESULTS: By using glucose values obtained by portable glucose meters instead of the results attained in the central laboratory, more women were diagnosed with gestational diabetes (33 vs 25) and the perinatal outcome was comparable. CONCLUSIONS: Obstetricians have to identify glucose intolerance during pregnancy in order to prevent macrosomia and its perinatal complications. These goals can be achieved with at least the same efficacy by performing the 100-g glucose tolerance test with a portable glucose meter as with current laboratory methods. Such an approach is time and resource saving and more convenient for both patients and health providers. PMID- 12747231 TI - Biovar diversity of Ureaplasma urealyticum in amniotic fluid: distribution, intrauterine inflammatory response and pregnancy outcomes. AB - OBJECTIVE: The objective of this study was to determine the distribution of two biovars of Ureaplasma urealyticum (parvo and T960) in human amniotic fluid and to examine whether the magnitude of the intrauterine inflammatory response and pregnancy outcomes are different between patients with microbial invasion of the amniotic cavity with "parvo biovar" and those with "T960 biovar". STUDY DESIGN: This cohort included 77 preterm singleton pregnancies (gestational age < 37 weeks) in whom U. urealyticum was detected from amniotic fluid using the polymerase chain reaction (PCR). Amniotic fluid was obtained by transabdominal amniocentesis. Amniotic fluid was cultured for aerobic and anaerobic bacteria as well as mycoplasmas. U. urealyticum was biotyped by PCR methods. Amniotic fluid inflammatory response was determined by amniotic fluid white blood cell count and interleukin-6 concentration. RESULTS: 1) The "parvo biovar" was detected in 82% (63/77) and "T960 biovar" was in 18% (14/77) of cases; 2) U. urealyticum was isolated by conventional culture method from amniotic fluid in 56% (35/63) of cases with positive for "parvo biovar" and in 50% (7/14) of cases with positive for "T960 biovar"; 3) There were no significant differences in the median gestational age at amniocentesis, gestational age at delivery, birth weight, amniotic fluid white blood cell count, amniotic fluid interleukin-6 concentration and the rates of clinical chorioamnionitis, histologic chorioamnionitis, funisitis and neonatal morbidity between patients in the two biovar groups. CONCLUSIONS: 1) The "parvo biovar" is more frequently isolated from amniotic fluid of preterm gestations than the "T960 biovar"; 2) Biovar diversity of U. urealyticum in amniotic fluid was not associated with different pregnancy outcome and magnitude of the intraamniotic inflammatory response. PMID- 12747232 TI - Comparative study on the expression of cytokine--receptor genes in normal and preeclamptic human placentas using DNA microarrays. AB - AIMS: To study the relationship between the expression levels of cytokine/receptor genes in placenta and the pathogenesis of pre-eclampsia. METHODS: The study was performed to compare the mRNA contents of cytokine (receptor) superfamily genes in placentas from 5 patients with pre-eclampsia and 5 strictly matched normal pregnancies. A complementary DNA microarray representing over 220 cytokine-associated genes was employed to complete the detection. RESULTS: It was shown that, among the 221 kinds of cytokine-associated genes, 162 of those including 22 interleukin/interleukin receptor genes presented with a difference of over two times in pre-eclamptic placentas compared to normal placentas. Most of the 22 interleukin/interleukin receptor genes were shown to be highly expressed in preeclamptic placenta, while the expression of IL-2 receptor (IL-2 R alpha, GenBank: X01057) gene in preeclamptic placenta was comparatively lower than that in normal placenta. Furthermore, some tumor necrosis factor (TNF)/receptor superfamily genes, including TNF (GenBank: X02910), TNF ligands (GenBank: U03398, U37518, AF053712, AF055872) and TNF receptors (GenBank: X60592, X63717, M83554, AF016266, AF016267, U81232) were also shown to be highly expressed in pre-eclamptic placenta. Besides interleukin and tumor necrosis factor (receptor) gene superfamily, the mRNA levels of another 39 cytokine and 15 cytokine receptor genes showed a two-fold difference between pre-eclamptic and normal placental tissues. Additionally, most of the genes were up-regulated in pre-eclamptic placenta. CONCLUSIONS: The up-regulation of cytokine-associated genes including interleukin and TNF (receptor) superfamily expression in placenta might be intensively related to the pathogenesis of pre-eclampsia. PMID- 12747233 TI - Chorioangioma--new insights into a well-known problem. I. Results of a clinical and morphological study of 136 cases. AB - AIMS: Chorioangiomas are rare hamartomatous lesions. Possible correlations between their occurrence and the progression of a pregnancy have been objects of discussions for quite some time. METHODS: In a retrospective study 22439 unselected placentas were examined for incidences of chorioangiomas, morphological features and accompanying clinical characteristics. RESULTS: Chorioangiomas occur in 0.61% of pregnancies, they are mainly microscopically small, and 55% of them are localized subchorial. The rate of their occurrence rises almost linearly with maternal age; chorioangiomas are found most often in women who are over 30 years old. Hypertension and diabetes are found more often in combination with chorioangiomas than they are in otherwise normal pregnancies. In 72% of all cases girls were born; in 33% we also observed malfunctions in the maturation processes of the placental parenchyma, in particular arrested and delayed maturation of the villi. Premature births occur approximately three times more often in chorioangioma pregnancies than in normal ones. Chorioangiomas are often found in primipara and twin pregnancies. PMID- 12747234 TI - Chorioangiomas--new insights into a well-known problem. II. An immuno histochemical investigation of 136 cases. AB - AIMS: Chorioangiomas are benign tumors of the hemochorial placenta. They are malformations or hamartomas, formed as a result of defective angiogenesis. They are of clinical importance due to their association with premature placental release and pre-eclampsia. METHODS: Since a link has been established in neoplasias between tumor growth and an increased expression of angiogenic growth factors, 136 samples of chorioangiomas and 136 samples of tumor-free placental tissue were examined in terms of proliferation rate and expression of the growth factors angiopoietin-1 and -2, the angiopoietin-receptor Tie-2, PDGF and the PDFG beta-receptor. RESULTS: The chorioangiomas exhibited differing proliferation rates, whereas tumor-free placental tissue barely proliferated at all. Angiopoietin expression was--morphologically--considerably higher within the chorioangiomas than in the comparison placentas; morphological amounts of the Tie 2 receptor were identical in all samples. Expression of PDGF and its receptor was the same for chorioangiomas and tumor-free placentas. CONCLUSIONS: According to this study and the current literature in the field of hamartomas and some neoplasia, we can assume that increased growth factor expression plays a role in the formation of chorioangiomas, since it stimulates proliferation in a wide variety of cell compartments. PMID- 12747235 TI - Impaired CD14-dependent and independent response of polymorphonuclear leukocytes in preterm infants. AB - Preterm newborn infants are especially susceptible to Gram-negative sepsis that is associated with a lethality of up to 40%. AIMS: We tested whether polymorphonuclear leukocytes (PMN) from preterm infants exhibit an impaired antibacterial response upon stimulation with lipopolysaccharide (LPS) from Escherichia coli when compared to full term newborns or adults. METHODS: We studied the effect of LPS on the expression of the surface proteins CD11b and CD14 and the secretion of elastase by PMN from preterm infants, term infants and adults ex vivo. RESULTS: We found a significantly reduced antibacterial activity of PMN from preterm infants upon stimulation with LPS as indicated by low surface expression of the adhesion molecule CD11b and the reduced secretion of PMN elastase. LPS-induced CD11b expression was dependent on binding of LPS to the surface protein CD14 as CD14 antibodies inhibited LPS dependent CD11b upregulation. Furthermore CD14 expression was lower on PMN from preterm infants than from adults. In addition, CD14 independent upregulation of CD11b in response to tumor necrosis factor (TNF-alpha), N-formyl peptides (FMLP) and phorbol ester (PMA) was impaired. CONCLUSION: PMN from preterm infants are distinctly hyporesponsive to LPS, which may explain the predisposition of these children to invasive disease due to gramnegative bacteria. PMID- 12747237 TI - WHO Multicentre Collaborative Networks for Severe Acute Respiratory Syndrome (SARS) diagnosis. PMID- 12747236 TI - The left portal vein is the watershed of the fetal venous system. AB - Critical fetal organs are preferentially supplied with oxygenated blood from the umbilical vein (UV) by way of the ductus venosus (DV). Under normal conditions a significant part of UV-blood flows steadily forward through the left portal vein (LPV). Blood flow through the LPV could reverse, however, in cases of absent or reversed endodiastolic flow in the umbilical arteries. We tested when fetal blood flow reversal occurs by studying 28 cases with pathological flow in the umbilical artery. In the LPV we observed normal nonpulsatile forward flow in 9 cases, pulsatile forward flow in 10 cases, and reversed flow in 9 cases. Reverse flow in the LPV correlated significantly with an elevated resistance index of the umbilical arteries. This reversal could have major physiological implications: Deoxygenated blood may be added via the LPV to the blood shifted through the DV and ultimately reach critical fetal organs. In extremis there could be a waterhose effect, whereby more blood flows through the DV than the UV that supplies it. The LPV is thus the watershed of the venous circulation of the fetus. PMID- 12747238 TI - WHO annual report on global TB control--summary. PMID- 12747239 TI - JCAHO announces upcoming shift to unannounced surveys. PMID- 12747240 TI - HIPAA prep continues after deadlines pass. PMID- 12747241 TI - Project hinges on top quality hospital data. PMID- 12747242 TI - Responding to customer concerns improves quality. Part 2. PMID- 12747243 TI - Pediatric case management--putting the pieces together. PMID- 12747244 TI - Reducing use of restraints on a rehabilitation unit. PMID- 12747246 TI - A conceptual map for studying long-term exercise adherence in a cardiac population. AB - The failure of people with coronary heart disease to adhere to prescribed programs of long-term exercise is a growing concern among healthcare professionals. The problem is attributed partly to not correctly identifying the specific exercise adherence behavior desired. There is a need in cardiac rehabilitation for different theoretical and practical approaches to the study of patient adherence to short-term and long-term exercise prescriptions. This article is a review of behavioral and cognitive models used to promote exercise in cardiac rehabilitation programs. Concepts are borrowed from these models to design a conceptual map intended to identify variables and relationships associated with exercise adherence behaviors. In this article, the Coronary Artery Disease Exercise Map (CADEM), an evolving conceptual map, addresses exercise behavior within a coronary population. However, this map may also be applied to people who have other chronic conditions. It is believed that the CADEM will provide researchers and healthcare providers with a framework with which to understand, promote, and evaluate adherence to exercise regimes within the rehabilitation population. PMID- 12747245 TI - Understanding how disability is defined and conceptualized in the literature. AB - A goal of rehabilitation nursing is to advance the care of persons with chronic disabling conditions by minimizing disability. There are two distinct perpectives in the literature about how to conceptualize disability. Definitions and models of disability that inform rehabilitation nursing practice are based on a functionalist perspective of illness and locate disability as a problem of individual functioning. Alternatively, there are definitions and models that have evolved from the civil rights and social justice perspectives, and that locate the problem of disability within a disabling society. Recent attempts to integrate these two perspectives are based on earlier rehabilitation models and the functionalist assumptions remain. Rehabilitation nursing research and practice based on either of these two perspectives is fragmented and incomplete. To know how to define and approach disability-related issues, it is important to understand how people living with disability perceive its influences on their lives. PMID- 12747247 TI - Are there different predictors of distinct exercise components? AB - Rehabilitation nurses often recommend to aging adults that they exercise. Despite the recommendations and the widespread evidence of its benefits, few older adults exercise. This study examined the predictors of distinct components of exercise behavior: exercise intensity, frequency per week, duration of sessions, and months per year. Social cognitive theory predictors of exercise were measured with previously developed instruments. Interviews were conducted with community dwelling elders (N = 147). The study constructs accounted for modest to moderate amounts of variance (41% of months per year, 35% of frequency, 35% of exercise intensity, and 20% of episode duration). Self-efficacy was the most important predictor of each construct, with Beta weights ranging from .23 to .53. Barriers significantly predicted months' per year, frequency per week, and exercise intensity, but not the duration of individual exercise sessions. Outcome expectancy predicted only exercise intensity. These findings support the importance of self-efficacy and perceived barriers for most components of exercise behavior. PMID- 12747248 TI - An interdisciplinary pediatric rehabilitation project in Vietnam: the Temple team experience. AB - Four faculty members (two nurses, one physical therapist, one occupational therapist) from the College of Allied Health Professions, Temple University in Philadelphia, PA, were invited to Vietnam in March 2001 to present an interdisciplinary workshop on cerebral palsy to the rehabilitation staff at a large hospital for children in Hanoi. The purpose was to present lectures on pediatric rehabilitation, to demonstrate techniques, to conduct clinical assessment rounds, and to increase the staff's knowledge about outcome evaluation. After adjusting to the cultural differences, the team worked with Vietnamese professionals in their respective fields. The goals of the workshop were met except that it was not possible to evaluate outcomes by customary methods. However, trainees indicated verbally that they understood the new materials and techniques. The workshop was part of an ongoing project in rehabilitation in Vietnam that is sponsored by Health Volunteers Overseas, a voluntary agency in Washington, DC. PMID- 12747249 TI - [Clinical evaluation of a test for plasma fibrin/fibrinogen degradation products (FDP) based on monoclonal anti-FDP antibody technology: an application for the scoring system of the disseminated intravascular coagulation (DIC) diagnostic criteria]. AB - Fibrin/fibrinogen degradation products(FDP) have been measured using serum samples which were specially prepared for the FDP test because of the usage of anti-human fibrinogen antibody for the assay. Since diagnostic criteria for DIC were established by the study group on thrombosis and hemostasis which is supported by the Japanese Ministry of Health and Welfare(JMHW), serum FDP assay have been used as standard methods to diagnose DIC in Japan. Recently, a reagent using an anti-human FDP monoclonal antibody was developed and this has enabled the use of plasma samples for FDP measurement. The comparability, especially of the DIC score, of a new assay, Latex test BL-2 P-FDP, using plasma samples with a conventional assay for serum was investigated. Two sets of DIC scores based on data from the two tests were compared and the correlation was high with 97.5% of the patients being diagnosed with the same DIC status. In four disease groups- DIC, thrombosis, leukemia and solid cancer--high comparability between the two tests was also shown and no significant difference was observed in the correlation coefficient and the slope coefficient between serum and plasma samples. To conclude, it is suggested that "Latex test BL-2 P-FDP" is applicable to the diagnostic criteria for DIC from JMHW without any difficulty. PMID- 12747250 TI - [Factors of the successful radiofrequency catheter ablation to the slow pathway in patients with uncommon atrioventricular nodal reentrant tachycardia]. AB - We studied the clinical features and factors of successful catheter ablation for common and uncommon atrioventricular nodal reentrant tachycardia(AVNRT). The study population consisted of 41 consecutive patients, 33 with common type AVNRT (16 males mean age of 57.8 years), and 8 patients with uncommon type AVNRT (4 males, mean age of 57.1 years). In all patients with common type AVNRT, the earliest atrial activation during tachycardia was recorded at the His bundle region. The effective ablation sites were located in the medial septal area and all cases were successfully ablated without complication. In patients with uncommon type AVNRT, dual pathway was observed in 5 patients(63%) and triple pathway in 3 patients(38%). The earliest atrial activation during tachycardias was recorded at the ostium of the coronary sinus. Radiofrequency ablation therapy was performed during sinus rhythm in 5 patients, and during tachycardia in 3 others. The effective ablation sites were located at the posterior septal area around the coronary sinus ostium. All cases were successfully ablated without any major complication, but one case had second-grade atrioventricular block after ablation. There were no significant differences between the common and uncommon type AVNRT cases with regard to the therapeutic success rate, the mean application number or the total energy applied. However, the successful ablation sites were different between the two groups. We concluded that radiofrequency catheter ablation would be effective in patients with both common and uncommon types AVNRT. Selective ablation at the site of the retrograde slow pathway exit was the most important factor for successful catheter ablation for uncommon type AVNRT. However, it should be performed only after careful analysis, taking into account the complex mechanism of uncommon type AVNRT. PMID- 12747251 TI - [Basic studies on mutation analysis of K-ras codon 12 by use of three-dimensional microarray system]. AB - A next-generation DNA microarray system, FD10 has been developed. It is based around the PamChip, a custom-made microarray, which consists of a solid three dimensional structure that facilitated the incorporation of probe molecules. We applied this microarray system on a detection of K-ras mutation at codon 12 in some cancer cell lines. The PCR products amplified by use of FITC labeled primers were applied onto probe-absorbed microarray. After hybridization, the signal was imaged by CCD camera and analyzed by the exclusive software. We confirmed the microarray results by PCR-SSCP and sequencing analyses. Ten, two and three out of 15 cell lines were homozygous for wild type allele, heterozygous for wild and mutant allele, and homozygous for mutant alleles, respectively. Signals hybridized with antisense probes were stronger than those with sense probes, without PSN1 cell line. The system had a good reproducibility. Essentially, the microarray results were consistent with PCR-SSCP and sequencing results. In conclusion, the FD10 microarray system was easy to operate and short to get results. It might be useful for a focused array applicable for specific purposes. The K-ras mutation detection system worked well and will be applied to clinical specimens soon. PMID- 12747252 TI - Development of peptide nucleic acid mediated polymerase chain reaction clamping (PMPC)--direct sequencing method for detecting lamivudine-resistant hepatitis B virus (HBV) variants with high sensitivity and specificity. AB - Reduced sensitivity of HBV to lamivudine, which causes a viral breakthrough during treatment, is attributed to mutations within the tyrosine-methionine aspartate(YMDD) locus in the reverse transcriptase(rt) domain of HBV polymerase, mainly a methionine(rtM204) substitution. The sensitive detection of such mutations before or early in treatment could assist in optimizing antiviral treatment. For this purpose, we developed peptide nucleic acid(PNA) mediated polymerase chain reaction(PCR) clamping(PMPC) with a PNA probe targeting the YMDD locus. We first tested this method for its sensitivity and specificity in detecting a mutant on HBV DNA standards consisting of serial copy number ratios of a known lamivudine-resistant, mutant clone with rtM204I(ATT) to a wild-type clone with rtM204(ATG). The sensitivity was 0.1 to 0.01% in the coexistence of wild-type clones and the specificity was guaranteed by direct sequencing of the products. We next applied this method to HBV DNA specimens extracted from serum from 4 chronic hepatitis B patients treated with lamivudine. Two of these exhibited a break-through of the HBV mutant with rtM204I(ATT), while the other 2 did not. Before treatment, all 4 patients showed HBV with rtM204I encoded by ATA. During treatment, HBV with the rtM204I(ATT) emerged in the 2 breakthrough patients more than 3 months before the breakthrough, whereas this and other known lamivudine-resistant viruses did not appear in the 2 non-breakthrough patients. Thus, our PMPC-direct sequencing method is highly sensitive and reliable for the early identification of lamivudine-resistant HBV that causes a viral breakthrough. PMID- 12747254 TI - [Information and consultation system in a commercial laboratory]. AB - We, the Japan Clinical Laboratories, Inc., have a department of information and consultation for our customers, medical institutions such as hospitals, clinics and so on. We collect samples from the customers and deliver the data from those samples back to them. Our department of information and consultation often receives various questions and complains from the customers. The contents of the questions and complains are about the clinical significance of the tests, the interpretation of the test results, and so on. We introduce them in this paper, in order to highlight the relations between commercial laboratories and their customers, and to point out the differences between commercial laboratories and the medical laboratories in hospitals. PMID- 12747253 TI - [Roles of the laboratory information division and laboratory technicians in changing medical circumstances]. AB - It is often voiced recently that the medical administration and medical environment are changing drastically. Hospital management in the future must face the challenges of improving efficiency and quality of medical care. Thus, promotion of team care is indispensable, and the clinical path and Nutrition support Team (NST) are rapidly spreading. We consider that the hospital laboratory must also implement and develop a Laboratory Information Division for more comprehensive exchange of information with clinical departments, and move beyond the laboratory by participating in team care. We have planned and organized the Kinki Laboratory Forum from 2000 as an opportunity for information exchange among laboratory technicians who aspire to establish the Laboratory Information Division and recognize the necessity of new actions for participation in team care. Its contents are presented. PMID- 12747255 TI - [Effective cooperation between clinical physicians and laboratory consultation division]. AB - With the increase in clinical examination items, and with the specialization of clinical medicine, physicians' requests for consultations on laboratory medicine is getting more evident. However, at present in many hospitals laboratory staff do not always make appropriate answers to the questions from physicians in various fields because the laboratories are divided in terms of examination fields. In our hospital, we established a consultation division responding to every inquiry from clinical staff with a full-time technologist. At present the division is functioning very well, suggesting that laboratory technologists must build close relations with clinical staff and that the technologist's task is very diverse, not confined to just analyzing samples. PMID- 12747256 TI - [Communication of useful information from laboratory physicians to clinical physicians]. AB - In recent years increasing importance has been placed on the role of hospital clinical research, such as the promotion of intra-laboratory human, material and informational resources, previously restricted to the laboratory, to the whole hospital system, and the appropriate usage of laboratory findings via common consultation systems, or specialized informational consultants in the clinical laboratory department. The volume of clinical laboratory information, which plays an important role in the decision-making process of routine clinical practices, is enormous for each individual hospital, and appropriate use of this information has a major influence on institutional clinical practice efficiency. In response to the need for the communication of useful laboratory information to clinical physicians, departments of laboratory research consultants have been organized in individual hospitals as a way forward. In the near future, laboratory physicians will play a leading role in the communication of research information from the viewpoint of EBLM (evidence-based laboratory medicine). From the work of these laboratory research consultants, it becomes possible to obtain relevant EBLM related information, such as frequently asked questions and opinions, from their users. By replying to these questions and opinions appropriately, laboratory research consultants can provide information that is both advantageous and useful, and which meets the needs of the clinical physician side. Effective communication of useful laboratory research information should not be restricted to either the laboratory physicians or the technicians; it is a job that needs the cooperation and teamwork of both sets of people. Also, they should always keep in mind that communication by itself is not sufficient; they should not assume the useful evaluation of information by the users, but rather ensure that they are presented with information that precisely meets their needs. PMID- 12747257 TI - [Newly-developed drug information services provided by hospital pharmacists]. AB - Modern health care is supported by many health care professionals. Hospital pharmacists are required to accept increasing responsibilities for appropriate medication use, moving into more important roles within the health care system. Accurate drug information is indispensable for appropriate medication use, however, drug information is not always sufficiently utilized by physicians, and thus it is suggested that there is a need for providing individualized drug information (one to one advice to physicians). Hospital pharmacists must have access to patient-specific health care information and must monitor drug therapy directly in order to provide individualized drug information. Hospital pharmacists, with proper access to patient-specific health care information, can promote safer prescribing practices by advising both patients and physicians, and so can reduce patients' risk for medication-related problems and improve the quality of their health care. Optimal drug therapy is the goal for every patient. Together, using their respective knowledge bases, pharmacists and physicians could provide optimal drug therapy, benefiting both patients and the health care system. PMID- 12747258 TI - [The proposed educational reforms of the undergraduate medical school curriculum and the postgraduate clinical training system]. AB - Along with the rapid advances in the fields of life science and technology, the knowledge that the medical students are expected to acquire has expanded in many areas, and also many subjects have been subdivided into special fields, thus resulting in a situation in which the knowledge that doctors obtain at medical school is quickly becoming less uniform. Therefore, the introduction of a standardized medical school curriculum and postgraduate training systems has been discussed over the past several years. The Departments of Education and Technology and Welfare and Labor have started to draw up new curricula and systems in order to ensure the quality of the medical doctors licensed to practice medicine in Japan. These reforms require appropriate changes in the educational programs for such fields as clinical pathology and laboratory medicine on both the undergraduate and the postgraduate levels. This workshop has been organized to introduce the findings of both original and advanced trials being performed by medical schools and educational hospitals with different backgrounds. At first, the progress of these reforms is summarized in order for the participants to obtain a better understanding of the undergoing reforms. PMID- 12747259 TI - [Compliance with the core curriculum of laboratory medicine (private school)]. AB - A model core curriculum for medical education was proposed in March 2001. Medical schools have to revise their curriculum according to the guideline. Showa University School of Medicine revised its curriculum of lectures and clinical practice for laboratory medicine according to the guideline. The GIO of the lecture at 4th grade was for the clinical practice at 5th grade. The GIO of the clinical practice at 5th grade was for the post-graduate clinical training. This revision is expected to improve education and result in better doctors. PMID- 12747260 TI - [The correspondence between the clinical laboratory medicine and the core curriculum of national universities]. AB - A model core curriculum for medical education is proposed to serve as the essential basis of material to be mastered at present and as a reference for curricula to be constructed by individual universities. The author carried out a questionnaire in departments of clinical laboratory medicine in the national universities in order to clarify the actual conditions of countermeasures of this education program for clinical laboratory medicine. As a result, 50% of universities satisfied this program's objectives and the remainder were insufficient. School hours are reduced by half. There were great differences in the methods of clinical laboratory training between universities. In the Toyama Medical and Pharmaceutical University, general clinical laboratory education is administered in the first semester of the 4th year. In clinical training, all the members should study compulsory fundamental laboratory technique in first semester of the 5th year and, in addition, 6th graders in the last 2 months should be able to choose an advanced clinical training course. To this basic curriculum addition of an elective curriculum will be required, providing individual or unique training programs based on student needs and the educational principles of each university. PMID- 12747261 TI - [Mandatory postgraduate medical training in Japan--present state of Nihon University as a private medical school]. AB - This is a time of considerable uncertainty about the future of the postgraduate medical education policy of the Japanese government. Strong and visionary academic leadership of laboratory physicians in private medical schools is needed. The medical schools must not only adapt to a changing health care system, but also maintain excellence in education, patient care, and clinical research. In Japan, tradition has it that the comparatively few faculty members at national medical schools are mostly promoted only on the basis of research in experimental medicine, therefore, young medical graduates are increasingly drawn to bench work or molecular medicine, not to clinical practice. Single-minded specialization tends to produce single track minds, which may lack balanced judgment in approaching the appropriateness of both investigation and management. For continuity of care and containment of costs, a year or two of general professional training after graduation preceded by a broad medical education is an invaluable investment. All medical graduates, whatever their intended or unintended final destination (even if not clinical), should spend more than six months in medicine and four or five months in surgery, at least half of each to be spent in the general disciplines, including responsibilities for acute emergency admissions. As certified laboratory physicians we must attempt to attract graduates into laboratory medicine by developing imaginative training programs including common laboratory procedures such as Gram's stain, Wright Giemsa stain and point of care testing at the patient's bedside or in ambulatory clinics, not only in central clinical laboratories. PMID- 12747262 TI - [Laboratory medicine in the obligatory postgraduate clinical training system- common clinical training program in the department of laboratory medicine in our prefectural medical university hospital]. AB - I propose a postgraduate common clinical training program to be provided by the department of laboratory medicine in our prefectural medical university hospital. The program has three purposes: first, mastering basic laboratory tests; second, developing the skills necessary to accurately interpret laboratory data; third, learning specific techniques in the field of laboratory medicine. For the first purpose, it is important that medical trainees perform testing of their own patients at bedside or in the central clinical laboratory. When testing at the central clinical laboratory, instruction by expert laboratory technicians is helpful. The teaching doctors in the department of laboratory medicine are asked to advise the trainees on the interpretation of data. Consultation will be received via interview or e-mail. In addition, the trainees can participate in various conferences, seminars, and meetings held at the central clinical laboratory. Finally, in order to learn specific techniques in the field of laboratory medicine, several special courses lasting a few months will be prepared. I think this program should be closely linked to the training program in internal medicine. PMID- 12747263 TI - [Postgraduate training program in laboratory medicine at a clinical teaching hospital]. AB - The Tenri Hospital resident system was introduced in 1976 and the training program for laboratory medicine began in 1982. Thus, the author proposes goals for the the future on the basis of experience. It is appropriate that trainees study emergency tests, blood transfusion and microbiology(particularly Gram's stain and sputum culture) as practical matters, and in addition to these, learn how to reply to consultations from physicians, learn the laboratory flow(so called laboratory system), and announce interpretations of laboratory data at reversed clinical pathological conference(R-CPC). The objectives of these training programs are to gain skills for appropriate laboratory utilization and interpretation, and develop communications and consultations with clinical pathologists and medical technologists. The key points of success in the training are close cooperation of the laboratory and teaching divisions. Particularly, cooperation with medical technologists is necessary, and it is essential medical practice for trainees because they will have to work with them in future. Finally it should be emphasized that there training has a limited effect because of the short duration. It is thus important to communicate and discuss clinical matters regularly in medicine. PMID- 12747264 TI - [Laboratory examinations performed in rural practice]. AB - Graduates from Jichi Medical School are obligated to work at rural clinics or hospitals, where most of them are the only medical doctor in the house. To understand how these graduates actually use laboratory examinations, what examinations they found most important in their practice, and when they were confident of their laboratory techniques, we designed a questionnaire to address these questions. Many respondents reported that their institutions had electrocardiographs, abdominal and/or cardiac ultrasonographs, urinalysis test paper, and portable blood glucose meters, and more than half of them reported having used these instruments without assistance in emergency situations. Moreover, a majority of the respondents said that they considered it important that a physician is able to use these instruments without the help of other staff members. Proficiency in many laboratory techniques was obtained and physicians were confident during their first postgraduate clinical practice. These responses clearly show the importance and usefulness of covering examination techniques and the principles of laboratory medicine in medical education and the first postgraduate clinical practice. PMID- 12747265 TI - [Releasing mechanisms of neurotransmitters]. PMID- 12747266 TI - Current issues of basic and clinical glucocorticoid research. PMID- 12747267 TI - Solar light effects on onset/relapses and circannual/circadian symptomatology in rheumatoid arthritis. PMID- 12747268 TI - Low-dose glucocorticoids in early rheumatoid arthritis: discordant effects on bone mineral density and fractures? AB - OBJECTIVE: To investigate the incidence of osteoporotic fractures and effects on bone of low-dose glucocorticoid (GC) monotherapy in a group of previously untreated patients with early active RA we performed a double blind, randomised, placebo-controlled clinical trial. The study duration was 2 years, with an open follow-up during the third year. Patients were randomly allocated to receive 10 mg prednisone or placebo. METHODS: Non-steroidal anti-inflammatory drugs (NSAIDs) were allowed in both groups. After 6 months sulphasalazine (2 gr daily) could be prescribed as rescue therapy in both groups. Except for 500 mg calcium supplement daily, no specific preventive measures were taken. This was a normal procedure at the time the study was designed (1989-1991). At the start of the study and every 6 months, X-rays of the twelfth thoracic and of all lumbar vertebrae were scored using the Kleerekoper method, and every year biochemical parameters of bone metabolism and bone mineral density (BMD, expressed in T-scores) and bone mineral content (BMC, expressed in g/cm) were assessed. RESULTS: In the prednisone group there was a higher incidence during the study of lumbar vertebral fractures than in the placebo group: 7 vs 4 respectively. This difference did not reach statistical significance however, probably because of the small numbers. One patient of the prednisone group suffered an osteoporotic fracture of the pelvis. In the 2-year study and the subsequent follow-up year, no other peripheral fractures were seen in either group. No significant changes from baseline in BMD and BMC of the hips were seen in either group during the study and the follow-up year. In the lumbar spine, BMD in the prednisone group decreased although not statistically significantly during the whole study. No correlation between changes in serum osteocalcin and BMD was observed. CONCLUSION: Low-dose prednisone monotherapy for patients with early active previously untreated RA seems to increase the risk of fractures not only by reducing the BMD but also by changes in bone strength and structure. PMID- 12747270 TI - Diacerein reduces the level of cartilage chondrocyte DNA fragmentation and death in experimental dog osteoarthritic cartilage at the same time that it inhibits caspase-3 and inducible nitric oxide synthase. AB - OBJECTIVE: The primary objective of this study was to evaluate the ex vivo therapeutic efficacy of diacerein and its active metabolite, rhein, on osteoarthritic (OA) cartilage chondrocyte DNA fragmentation and death in the experimental canine model of OA. The study also aimed to explore the effect of the drug on the level of important factors involved in this phenomenon, i.e., caspase-3 and inducible nitric oxide synthase (iNOS). METHODS: OA knee cartilage was obtained from dogs that had received surgical sectioning of the anterior cruciate ligament (ACL) and were sacrificed 12 weeks after surgery. Cartilage explants were cultured in the presence or absence of therapeutic concentrations of diacerein (20 micrograms/ml) or rhein (20 micrograms/ml). Cartilage specimens were stained for TUNEL reaction and immunostained using specific antibodies for active caspase-3 and iNOS. Morphometric analyses were also performed. RESULTS: In OA cartilage specimens, a large number of chondrocytes in the superficial layers stained positive for TUNEL reaction. Treatment with therapeutic concentrations of diacerein (20 micrograms/ml) or rhein (20 micrograms/ml) significantly reduced the level of chondrocyte DNA fragmentation to about the same extent in both treatment groups (P < 0.006, P < 0.002, respectively). The levels of caspase-3 and iNOS in cartilage explants were also significantly decreased (caspase-3, diacerein P < 0.04; caspase-3, rhein P < 0.0003; and iNOS, rhein P < 0.009, respectively) when compared to the control group. CONCLUSIONS: This study shows that diacerein/rhein can effectively reduce the level of OA chondrocyte DNA fragmentation and death under the present experimental conditions. This effect is mediated by a decrease in the level of caspase-3 expression, which could possibly be related in part to the reduced level of iNOS and secondarily to NO production. These findings provide additional new information about the mechanisms of action of diacerein on the progression of OA. PMID- 12747269 TI - Seasonal distribution of relapse onset in rheumatoid arthritis and spondyloarthropathy: the possible effect of the solar factor. AB - BACKGROUND AND OBJECTIVE: The seasonal effect on the relapse of rheumatoid arthritis and spondyloarthropathies is still unclear. To assess the seasonal distribution of relapse onset in rheumatoid arthritis (RA) and spondyloarthropathy (SpA) and its association with solar factors. METHODS: The monthly distribution of relapse onsets during the years 1998-2000 was retrospectively chart reviewed in 364 patients. In 1998 a total of 131 patients were studied; 60 with seropositive (sp) RA, 30 with seronegative (sn) RA and 41 with SpA; 113 patients in 1999: 44 with spRA, 38 with snRA and 31 with SpA; 120 patients in 2000: 56 with spRA, 38 with snRA and 26 with SpA. All of them were treated in the Department of Rheumatology, which serves the population of northwestern Israel. Solar activity was analyzed according to the "Solar Terrestrial Activity Report Charts 1998-2000". The Central Israel Bureau of Statistics provided the sun global radiation data. Data was assessed during the summer (April-September) and winter (January-March, October-December). The correlation between the monthly distribution of disease relapses and solar factors was measured (SPSS-10 for WIN). RESULTS: Relapses in spRA patients occurred mostly during the summer months with peak activity during the month of July 2000. Single monthly peaks of spRA relapse onset were noted in January 1998 1999 and April 1998 and for snRA in January 1998 and June 2000, but there were no seasonal differences for spRA, snRA and SpA in 1998-1999 and for snRA and SpA in 2000. Relapses in spRA patients were associated with a summer bias of increased solar activity and global solar radiation in 2000 compared with lower peak solar activity in 1998-1999. Furthermore, in 2000 we found a significant correlation of the spRA monthly relapse count to solar activity (p = 0.005) and global sun radiation (p = 0.048) unlike snRA and SpA. No above-mentioned association and correlation was noted in 1998-1999. We revealed mild negative correlation (p = 0.046) of SpA relapse count only to peak solar flux (PSF) by analysis of data for 1998-2000 as one united group. CONCLUSIONS: Relapses were more frequent during the summer of 2000 (May-June-July) in spRA but not in snRA and SpA. The reasons are still unclear. No seasonal differences were observed in 1998-1999. Enhanced solar activity in summer-2000 compared with 1998-1999 may be inferred to be the proposed cause but coincidence may occur as well. Outbreak in RA and SpA was not registered despite increased peak solar activity in 2000. We observed mild evidence of reciprocal relation between SpA relapsing and solar activity during 1998-2000. Solar and any other possible contributory factors remain still to be elucidated. PMID- 12747271 TI - Atlantoaxial disorders in rheumatoid arthritis associate with the destruction of peripheral and shoulder joints, and decreased bone mineral density. AB - OBJECTIVE: To evaluate whether cervical spine changes are associated with the destruction of shoulder or peripheral joints and with bone mineral density (BMD) in patients with long-term RA. METHODS: An inception cohort of 67 patients with seropositive and erosive RA were followed up for 20 years. Cervical spine, shoulder, hand and foot radiographs, and the BMD of the lumbar spine and femoral neck were evaluated. RESULTS: A positive relationship was detected between the occurrence of atlantoaxial disorders and the destruction of both shoulder (p < 0.001) and peripheral (p = 0.001) joints. In addition, the severity of anterior atlantoaxial subluxation and atlantoaxial impaction positively correlated with the grade of destruction in the evaluated joints. Furthermore, patients with atlantoaxial disorders presented decreased BMD of the femoral neck (p = 0.019). The occurrences of subaxial subluxations (SAS) and subaxial disc space narrowings only associated with higher onset age of RA. CONCLUSIONS: Patients with severe RA and osteoporosis have an increased risk for atlantoaxial disorders. The co existence of shoulder destruction and cervical spine disorders makes the differential diagnosis of shoulder and neck pain challenging. PMID- 12747273 TI - Topical diclofenac patch in patients with knee osteoarthritis: a randomized, double-blind, controlled clinical trial. AB - OBJECTIVE: To assess the efficacy and safety of a diclofenac hydroxyethylpyrrolidine (DHEP) patch in the treatment of symptomatic osteoarthritis (OA) of the knee joint. METHODS: A double-blind, randomised, placebo-controlled trial was carried out on 103 outpatients for 2 weeks. The main efficacy parameters were spontaneous pain and Lequesne's Index. Secondary endpoints were walking time over a standard distance, global assessment of efficacy and tolerability, and paracetamol consumption. RESULTS: The active treatment group showed a significant improvement in pain, Lequesne's Index, and the physician's and patient's global assessment of efficacy. For these parameters the difference between groups was statistically significant in favour of the DHEP patch. Adverse reactions were seen in a small number of probands and were similar in both groups. CONCLUSIONS: The results of this trial suggest that the DHEP patch appears to be an effective and safe treatment for patients suffering from symptomatic knee OA. PMID- 12747272 TI - Dendritic cells co-localize with activated CD4+ T cells in giant cell arteritis. AB - OBJECTIVE: Giant cell arteritis (GCA) is a vasculitis predominantly affecting medium- and large-sized arteries. Recent data show the co-localization of dendritic cells and Chlamydia pneumoniae in vascular biopsies from GCA patients. Here we define the topographical relation of dendritic cells and these activated T-cells to determine the antigen presenting cell in GCA, and to examine several auxiliary biochemical and genetic aspects relating to the role of bacteria such as C. pneumoniae in eliciting GCA. METHODS: 18 paraffin-embedded temporal artery biopsy specimens from 14 patients with GCA that were PCR-positive for C. pneumoniae were examined by two-color immunohistochemistry for the topographical relationship between dendritic cells and activated T-cells. In addition the presence of GTP-binding proteins. Tumor necrosis factor alpha (TNF alpha), and Toll-like receptor 4 (TLR4) was investigated. 15 temporal artery specimens from 10 patients without GCA served as controls. RESULTS: In all GCA specimens, dendritic cells co-localized in the immediate vicinity of activated CD4+ Talin expressing T cells, and these were predominantly found in granulomatous infiltrates. Confocal microscopy confirmed the cell-cell contact of dendritic cells with activated T cells. Results further showed that RhoA and Rac1 were predominantly present in the region of granulomatous infiltrates. TNF alpha production and expression was found in dendritic cells and macrophages, predominantly in granulomatous infiltrates and in endothelial cells of the vasa vasorum dispersed in the adventitial and medial layers of the temporal artery. No control specimens showed TNF alpha expression. More than 95% of dendritic cells were positive for TLR4; macrophages and endothelial cells localized in the adventitia showed TLR4 production. CONCLUSIONS: The immediate co-localization of dendritic cells and activated T cells indicate a high probability that the former represent the antigen presenting cells in GCA. In addition, because of the presence of Rho A and Rac1 in the granulomatous infiltrates, we speculate that they provide the right environment for cell-cell contact and adhesion, and that they may promote the internalization of bacteria. TNF alpha is expressed at high levels in the granulomatous infiltrates of temporal artery specimens from patients with GCA. Since TLR4 is produced in the same cell types, and predominantly in the adventitial layer of the temporal artery, we suggest that these receptors are coupled to signal transduction pathways that control TNF alpha expression. PMID- 12747274 TI - Systemic sclerosis is not associated with clinical or ambulatory blood pressure. AB - BACKGROUND: Raynaud's phenomenon is often the first symptom and occurs eventually in more than 95% of patients with systemic sclerosis (SSc). Angiographic studies disclose narrowing and obstruction of the digital arteries, which on autopsy histologic study show prominent subintimal connective tissue proliferation without inflammation, as well as adventitial fibrosis. It is also known that primary cardiac problems include pericarditis, left ventricular or biventricular failure, serious supraventricular or ventricular arrhythmias emerge in patients with SSc. It is not known if these patients present hypertension or hypotension and which parameter of the ambulatory blood pressure may influence such a disease course. METHODS: A total of 85 subjects underwent clinical blood pressure (BP) readings, 24-hour ambulatory BP monitoring, left ventricular assessment by echocardiography and measurement of intima media thickness (IMT) of the right left internal carotid arteries (RICA and LICA) and right-left common carotid arteries (RCCA and LCCA). The population consisted of 40 subjects with SSc according to the criteria of the American College of Rheumatology (SCL-group) who were not receiving any antihypertensive treatment and 45 healthy volunteers (control group). The two groups did not differ in age. RESULTS: Clinical systolic and diastolic blood pressure, clinical heart rate, mean 24 h systolic blood pressure, SD systolic blood pressure, mean 24 h diastolic blood pressure, SD 24 h diastolic blood pressure, mean 24 h heart rate, SD 24 h heart rate, pulse pressure 24 h, serum glucose, cholesterol, triglycerides, HDL, LDL, creatinine, urea, potassium and natrium did not statistically significant differ between the two groups. Furthermore, the left ventricular mass/BSA and IMT of both carotid arteries did not show a statistically significant difference between the groups. CONCLUSION: Systemic sclerosis is not associated with clinical blood pressure or the parameter of 24 h blood pressure monitoring. PMID- 12747275 TI - MMP-8 is only a minor gene product of human adult articular chondrocytes of the knee. AB - OBJECTIVE: The initial degradation of collagen fibrils during osteoarthritic cartilage destruction depends on the cleavage at the collagenase site, for which there exist three major candidate enzymes: collagenase 1 (MMP-1), collagenase 2 (MMP-8), and collagense 3 (MMP-13). The objective of this study was to determine the quantitative expression as well as distribution levels in normal and osteoarthritic cartilage and synovium and in cultured articular chondrocytes with and without stimulation by Il-1 beta. METHODS: Conventional and online PCR technology and immunohistochemistry were used to determine MMP-8 expression levels on the mRNA and protein level. RESULTS: Whereas conventional PCR analysis could demonstrate the presence of MMP-8 mRNA in normal and osteoarthritic chondrocytes, online quantitative PCR showed that only very minor amounts of MMP 8 mRNA expression is found in articular chondrocytes in vivo (and in vitro) and that there is no significant upregulation in osteoarthritic cartilage in vivo nor by Il-1 beta in vitro. The in vivo results were confirmed by the absence of significant protein staining with monoclonal antibodies for MMP-8 in normal and osteoarthritic chondrocytes. CONCLUSIONS: The presented results confirm the presence of a very minor MMP-8 expression by articular chondrocytes, but clearly question the hypothesis that MMP-8 is a major cartilage matrix degrading protease and is involved in enhanced cartilage matrix breakdown in osteoarthritic cartilage degeneration or by Il-1 beta stimulation in vitro. PMID- 12747277 TI - Prevalence of Sindbis-related (Pogosta) virus infections in patients with arthritis. AB - OBJECTIVE: To determine the role of Pogosta virus as a triggering infection in non-specific arthritis. METHODS: Serum samples of 142 patients with acute arthritis were screened for the evidence of Pogosta virus infection. Serological tests for Chlamydia trachomatis, salmonella, parvovirus B19, and Borrelia burgdorferi were also carried out. As verified later, 78 of the patients had rheumatoid arthritis and 63 seronegative poly- or oligoarthritis, while one had systemic lupus erythematosus. RESULTS: In the early stage of the joint symptoms 4 patients with rheumatoid arthritis, 1 with seronegative polyarthritis and 1 with systemic lupus erythematosus had recent Pogosta virus infection. Four of them had probably had Pogosta disease at the time of the onset of arthritis. In 11 patients with a diagnosis of seronegative arthritis, serological evidence of preceding infection due to salmonella or Chlamydia trachomatis was found, strongly suggesting classical reactive arthritis in these cases. CONCLUSIONS: Our study suggests that also a Sindbis virus infection may be associated both to an acute joint inflammation as a part of Pogosta disease or chronic arthritis. At present, this possibility still needs further research. PMID- 12747276 TI - Autoantibodies and human immunodeficiency viruses infection: a case-control study. AB - OBJECTIVE: To determine the prevalence of organ-specific and non-specific autoantibodies in HIV-infected patients. DESIGN: A multicentric collaborative case-control study including 105 HIV patients and 100 sex- and age-matched HIV negative healthy volunteers. METHODS: Antinuclear, anti-ds DNA, anti-histone, anti-Sm, rheumatoid factor(IgM), anti-beta 2 glycoprotein 1, antineutrophil cytoplasmic, anti-LKM1, anti-LCA1, anti-gastric parietal cell, antiplatelet, anti intermediate filament, anti-mitotic spindle apparatus, anti-Golgi, anti-ribosome and anti-thyroid autoantibodies were screened in six European laboratories. RESULTS: Only IgG and IgM anticardiolipin, IgG antiplatelet, anti-smooth muscle and anti-thyroglobulin antibodies were statistically more frequent in HIV patients. There was no correlation with the numbers of CD4+ cells except in the case of anti-smooth muscle antibodies. We were unable to find specific autoantibodies such as anti-ds DNA, anti-Sm, AMA, anti-LKM1, anti-LCA1 or anti beta 2 GP1 antibodies in these patients. CONCLUSIONS: Our results indicate that the autoantibody profile of HIV infections is comparable to those of other chronic viral infections. HIV does not seem to be more autoimmunogenic than other viruses. PMID- 12747279 TI - Link between anti-CD36 antibodies and thrombosis in the antiphospholipid syndrome. AB - OBJECTIVE: Some studies have previously suggested the involvement of antibodies directed against CD36 (anti-CD36) in the pathogenesis of thrombosis. The aim of this study was to evaluate the prevalence of anti-CD36 in patients with antiphospholipid antibodies (aPL) and its relationship with thrombosis. METHODS: Anti-CD36 were tested using an indirect MAIPA assay in 62 patients with autoimmune aPL but without SLE; there were 38 with and 24 without thrombosis. Nineteen patients with thrombosis served as an aPL(-) control group and 58 healthy subjects as the normal control group. RESULTS: 15 of 62 aPL patients (24.2%) but only 1 of 58 (1.7%) normal controls had anti-CD36 (p < 0.0005). As compared to normal controls, the prevalence of anti-CD36 was significantly higher in aPL patients with (26.3%, p < 0.0005) or without thrombosis (20.8%, p < 0.01). Anti-CD36 were significantly more frequent in aPL patients with thrombosis than in thrombosis aPL(-) subjects (26.3% vs 0%, p = 0.02). The presence of anti-CD36 seems to be more frequent in aPL patients with recurrent thrombosis than in those with a single episode (36.8% vs 15.8%). CONCLUSION: The presence of anti-CD36 is highly prevalent in patients with autoimmune aPL with a trend to being more frequent in patients with recurrent episodes of thrombosis. PMID- 12747278 TI - Glucocorticoid receptor up-regulation in early rheumatoid arthritis treated with low dose prednisone or placebo. AB - OBJECTIVE: Low or medium dose prednisone in early rheumatoid arthritis (RA), albeit with significant variation in clinical efficacy, reduces the progression of joint damage. The glucocorticoid receptor (GR) number in peripheral mononuclear cells (PBMC) might be helpful to predict which patients will respond to low or medium dose prednisone and therefore do not or will not need higher doses. With this in mind we determined in a double blind, placebo controlled study at baseline and yearly the GR number in PBMC. METHODS: Eighty-one early RA patients (disease duration less than one year) were included. All patients fulfilled the ACR criteria and were disease modifying antirheumatic drugs (DMARD) and glucocorticoid-naive. They were randomly assigned to treatment with 10 mg prednisone daily or placebo. From all patients disease activity (CRP, number of tender and swollen joints), the radiological joint score, bone mineral density, and the GR number in PBMC were measured annually. RESULTS: In females the GR number was up-regulated over time in both the prednisone and the placebo group. The same trend was observed in males. No correlations were found between the GR number in the prednisone users at the start of their treatment and changes in radiological scores or bone density after 2 years of treatment. No correlations were found between the GR number at the start and the clinical characteristics after a follow-up of 2 years. CONCLUSION: The GR number in the PBMC of early RA patients did not predict which patients would be prednisone responders based on clinical or radiological parameters. However, the up-regulation of the GR number in PBMC in early RA patients towards the GR number of healthy subjects during the first two years of their disease course seems to reflect a recovery or compensatory mechanism as a response to an ongoing inflammatory process. This recovery may be not enough to efficiently control the inflammatory situation. PMID- 12747280 TI - Impaired expression of erythrocyte glycosyl-phosphatidylinositol-anchored membrane CD59 in patients with psoriatic arthritis. Relation to terminal complement pathway activation. AB - OBJECTIVE: Complement-mediated injury is regulated by many factors; among these CD59 has been identified as a widely distributed glycoprotein that inhibits membrane C5b-9 (terminal complement component) formation. The aim of the study was to assess erythrocyte CD59 expression in patients with psoriatic arthritis in order to understand the role of CD59 in the pathogenesis. METHODS: Washed erythrocytes from 50 patients with psoriatic arthritis, 8 with cutaneous psoriasis and 24 healthy subjects were incubated with monoclonal anti-CD59 antibody followed by a second FITC conjugated antibody and fluorescence intensity analysed by FAC-Scan flow cytometer to assess their CD59 membrane expression. SC5b-9 levels were measured in the plasma by ELISA and results compared with CD59 values. Immune complexes, complement C3 and C4 and rheumatoid factor were also determined. RESULTS: Impaired expression of erythrocyte membrane-anchored CD59 was found in patients with psoriatic arthritis; the lowest levels were seen in active patients (p < 0.01). Increased SC5b-9 was seen in the plasma of patients with active disease. An inverse correlation was also found between plasma C5b-9 and the CD59 expression levels (r = -0.81, p < 0.001). CONCLUSION: The low CD59 expression on erythrocytes from patients with psoriatic arthritis may be an index of a low tissue CD59 expression. This impairment could facilitate the activation of complement pathway and increase the risk for arthritis. Membrane attack complex formation in deficient membrane bound CD59 may also exacerbate synovial cell injury and inflammation. PMID- 12747281 TI - Rofecoxib exerts no effect on platelet plug formation in healthy volunteers. AB - OBJECTIVE: Although rofecoxib has very high selectivity for cyclo-oxygenase 2 (COX-2), supratherapeutic rofecoxib concentrations (> 1000 mg) inhibit purified human COX-1 in vitro and TXB2 formation in vivo. It is therefore possible that higher doses of rofecoxib may affect platelet function. This could be important if rofecoxib is given to thrombocytopenic patients. In these cases, already moderate inhibition of platelet function could precipitate bleeding complications. We therefore set out to investigate the influence of rofecoxib on platelet function in healthy volunteers. METHODS: We set up a balanced randomised, double-blind, placebo-controlled, two way cross-over study. Peripheral blood was withdrawn from 42 healthy volunteers before and 3 hours after intake of 50, 250, 500 mg of rofecoxib or placebo (n = 14 per group). Platelet function was assessed by a platelet function analyzer (PFA-100) which measures collagen-epinephrine induced closure time (CEPI-CT) under shear stress. RESULTS: CEPI-CT increased by 14% (p = 0.002) and 11% (p = 0.003) three hours after intake of placebo and rofecoxib at dosages of up to 500 mg, respectively. The increase in CEPI-CT versus baseline was not significantly different in the placebo period compared with the active treatment periods (n = 42, p > 0.05). CONCLUSIONS: Rofecoxib does not impair platelet function. Thus, rofecoxib appears to be a valuable analgetic and antipyretic agent in the therapy of patients at risk for bleeding. PMID- 12747282 TI - Biochemical markers of bone turnover, serum levels of interleukin-6/interleukin-6 soluble receptor and bisphosphonate treatment in Erdheim-Chester disease. AB - Erdheim-Chester disease (ECD) is a rare non-Langherans form of histiocytosis characterized radiologically by symmetrical sclerosis of the metaphysis and the diaphysis of long tubular bones. Macrophages are potent interleukin-6 (IL-6) producers and elevated IL-6 serum levels have been described in pathological conditions characterized by increased bone resorption. In a patient with ECD, during the acute phase of the disease we found high serum levels of IL-6 and IL-6 soluble receptor (sIL-6R) and high levels of bone turnover markers. After 5 years of combination therapy with oral prednisone and intravenous clodronate a significant reduction in the above mentioned biological parameters was seen. We suggest that the systemic disorders present in ECD could be related to the high serum levels of IL-6 and sIL-6R. We also propose the use of bisphosphonates in the clinical management of ECD. PMID- 12747283 TI - Paravertebral leiomyosarcoma mimicking a chronic ongoing inflammatory process. AB - Leiomyosarcomas usually present with symptoms associated with the tumor site or as painless soft tissue masses. We report the case of a young woman with spiking fever and elevated acute reaction proteins for months, in the context of a paravertebral high grade leiomyosarcoma. PMID- 12747284 TI - Non-contrast computed tomography-guided intra-articular corticosteroid injections of severe bilateral hip arthritis in a patient with ankylosing spondylitis. AB - Progressively destructive hip arthritis is a common complication of patients with ankylosing spondylitis. Palliative intra-articular corticosteroid hip injections are helpful in younger patients who wish to delay joint replacement surgery. Non contrast computed tomography (CT)-guided intra-articular procedures can potentially minimize radiation exposure compared to fluoroscopy-guided techniques. We describe a CT-guided technique for intra-articular corticosteroid hip injections for severe bilateral hip arthritis which avoids the use of radiologic contrast by documenting the presence of a pre-injection air bubble underneath the fibrous capsule. Significant reduction in symptoms within 48 hours and at 6 weeks post-injection confirmed the successful outcome of this technique. PMID- 12747285 TI - Molecular differences in anticytokine therapies. AB - Biologic agents that inhibit proinflammatory cytokines have made a profound impact on the treatment of rheumatoid arthritis (RA). Of the agents that are currently approved by the US Food and Drug Administration (FDA) for this indication, etanercept and infliximab neutralize tumor necrosis factor (TNF), and anakinra inhibits interleukin-1 (IL-1). Adalimumab, which was just recently approved by the FDA, is also a TNF inhibitor. Despite their common ability to inhibit cytokine bioactivity, the molecular structures and mechanisms of action of these biologic agents are significantly different. The TNF-binding moiety of etanercept is derived from soluble TNF receptor subunits. Infliximab is a chimeric (mouse-human) monoclonal antibody to TNF, while adalimumab is a fully human anti-TNF monoclonal antibody. Anakinra has yet another mechanism of action: it is an IL-1 receptor antagonist. The molecular characteristics of these agents may be relevant to clinical efficacy and safety. These agents are still relatively new: to date, the longest reporting time is 5 years, for etanercept. Additional long-term data will be required to determine the relative benefits and drawbacks of different molecular characteristics in these anticytokine agents. PMID- 12747286 TI - Hyperhomocysteinemia in children with juvenile idiopathic arthritis is not influenced by methotrexate treatment and folic acid supplementation: a pilot study. AB - OBJECTIVE: Our first objective was to compare plasma total homocysteine (tHcy) concentrations in juvenile idiopathic arthritis (JIA) patients requiring methotrexate (MTX) treatment and healthy children. Our second aim was to evaluate the influence of low-dose (10-15 mg/m2/week) MTX treatment combined with folic acid supplementation (1 mg/d) or placebo on tHcy concentrations in JIA patients. METHODS: In 17 JIA patients and 17 age- and sex-matched healthy children, baseline tHcy concentrations were measured. When MTX treatment was initiated, JIA patients were randomly assigned to folic acid 1 mg/d/p.o. followed by placebo (8 weeks each) or vice versa. Blood samples for measurement of tHcy, vitamin B6, B12 and folate were taken after 4 weeks, 12 weeks and 20 weeks of treatment. RESULTS: 1) In the healthy children the mean tHcy concentration was 6.3 +/- 1.68 mumol/l as compared to 9.99 +/- 5.17 mumol/l in JIA patients (p < 0.04). At baseline, 5/17 JIA patients had tHcy concentrations > 10.5 mumol/l, the 99th percentile for teenagers. 3/5 patients even exceeded the upper normal level for adults (tHcy > or = 15 mumol/l). MTX treatment did not result in a significant increase of tHcy and folic acid supplementation had no significant impact on tHcy levels. CONCLUSION: This pilot study shows that patients with JIA requiring MTX treatment have significantly elevated baseline plasma tHcy concentrations compared to age- and sex-matched healthy controls. No significant impact of MTX and folate supplementation on tHcy concentration was found. PMID- 12747287 TI - Differential peptide binding motif for three juvenile arthritis associated HLA-DQ molecules. AB - OBJECTIVE: In the oligoarticular subgroup of juvenile idiopathic arthritis, a strong association has been found with the expression of human leukocyte antigen class II molecules HLA-DQA1 *0401-DQB1*0402 and DQA1*0501-DQB1*0301, whereas DQA1*0501-DQB1*0201 is neutral and DQA1 *0201-DQB1*0201 protective. A presentation of different peptides by these DQ alleles would support their role in the disease process. METHODS: Using a synthetic nonapeptide library, a peptide binding motif was determined for the associated DQA1*0501-DQB1*0301 molecule and compared to the neutral and the protective DQ molecules. RESULTS: A differential motif for the three molecules could be deduced, suggesting that peptides preferentially binding to the associated vs. the neutral/protective DQ-molecules are mutually exclusive. CONCLUSION: These results imply a role for differential peptide presentation in the pathogenesis of oligoarthritic JIA. The search for peptides initiating the disease process might be facilitated which could then lead to therapeutical interventions. PMID- 12747288 TI - Occurrence of chronic inflammatory rheumatic diseases among parents of multiple offspring affected by juvenile idiopathic arthritis. AB - OBJECTIVE: The rarity of reports on extended multiplex families points out that the genetic component in juvenile idiopathic arthritis (JIA) might not be particularly strong. Our objective was to determine the frequency of chronic inflammatory rheumatic diseases among the parents who had two or more offspring affected by JIA. METHODS: During the last 17 years patients with JIA treated at the Rheumatism Foundation Hospital in Heinola and their parents have been systematically asked about the familial occurrence of rheumatic diseases. A total of 45 families with more than one sibling affected by JIA were found among about 2,300 JIA cases. In these "multicase families", 9 parents from 8 families also had a diagnosis of chronic inflammatory rheumatic disease. Their case histories were studied. RESULTS: Four of the parents had had JIA (one subsequently developed ankylosing spondylitis), and 4 had rheumatoid factor-negative chronic arthritis (one had also had chronic iritis since the age of 10, resembling that seen in JIA). Three of them had features of JIA and only one met the classification criteria for rheumatoid arthritis. One had ankylosing spondylitis. CONCLUSIONS: Since the expected number of JIA cases among the 90 parents was about 0.2, there was drastic increase in JIA frequency among the parents in families with multiple offspring also affected by JIA. These results suggest that JIA susceptibility genes may likely be clustered in these families. PMID- 12747289 TI - Familial Mediterranean fever and Behcet's disease. PMID- 12747290 TI - Salivary gland lymphocytic infiltrates and Helicobacter pylori serology in anti SSA/Ro positive patients in Italy. PMID- 12747291 TI - IgA nephropathy associated with eosinophilic fasciitis: report of a case. PMID- 12747292 TI - N-acetylcysteine and vitamin E: an in vitro study of their effect on homogentisic acid polymerization. PMID- 12747293 TI - Sacroiliitis as a manifestation of Hodgkin's disease in young females. PMID- 12747294 TI - Detection of Borrelia burgdorferi by species-specific and broad-range PCR of synovial fluid and synovial tissue of Lyme arthritis patients before and after antibiotic treatment. PMID- 12747295 TI - Efficacy of thalidomide in refractory adult Still's disease: a new case report. PMID- 12747297 TI - Inflammatory polyenthesopathy in a patient with X-linked osteomalacia. PMID- 12747296 TI - A single infusion of infliximab increases the serum endostatin level in patients with rheumatoid arthritis. PMID- 12747298 TI - Anti-Golgi antibodies in adult Still's disease. PMID- 12747300 TI - Epithelial cell-derived neutrophil activator-78 levels in children with familial Mediterranean fever. PMID- 12747299 TI - Drug-induced systemic lupus erythematosus associated with etanercept therapy in a child with juvenile idiopathic arthritis. PMID- 12747301 TI - High risks. PMID- 12747302 TI - Applicability of the International Classification of Nursing Practice (ICNP) in the areas of nutrition and skin care. AB - PURPOSE: To evaluate completeness, granularity, multiple axial content, and clinical utility of the beta version of the ICNP in the context of standardized nursing care planning in a clinical setting. METHODS: An 35-bed acute care ward for infectious diseases at a Swedish university hospital was selected for clinical testing. A convenience sample of 56 patient records with data on nutrition and skin care was analyzed and mapped to the ICNP. FINDINGS: Using the ICNP terminology, 59%-62% of the record content describing nursing phenomena and 30%-44% of the nursing interventions in the areas of nutrition and skin care could be expressed satisfactorily. For about a quarter of the content describing nursing phenomena and interventions, no corresponding ICNP term was found. CONCLUSIONS: The ICNP needs to be further developed to allow representation of the entire range of nursing care. Terms need to be developed to express patient participation and preferences, normal conditions, qualitative dimensions and characteristics, nonhuman focus, and duration. PRACTICE IMPLICATIONS: The practical usefulness of the ICNP needs further testing before conclusions about its clinical benefits can be determined. PMID- 12747303 TI - Environmental nursing diagnoses: a proposal for further development of Taxonomy II. AB - PURPOSE: To propose further development of environmental diagnoses and to offer recommendations for expanding Taxonomy II to include more diagnostic labels that encompass the environmental domain. SOURCES: Literature in the disciplines of nursing, biology, toxicology, public health, sociology, and anthropology. DATA SYNTHESIS: Nurses need language to describe the human responses of individuals, families, communities, and global society to environmental health threats. CONCLUSIONS: New environmental diagnoses will lead to refinement of language that describes the contribution of nursing to an emerging international and community health priority. PRACTICE IMPLICATIONS: Environmental diagnostic labels will allow nurses to name responses and plan interventions that respond to instances or risks of exposure to threats from the physical and sociocultural environment. PMID- 12747304 TI - End-of-life care legislation signed into law. PMID- 12747305 TI - Labor laws unveiled. PMID- 12747306 TI - Assisted living issues and concerns in Michigan. PMID- 12747308 TI - Can diabetes cause depression. PMID- 12747307 TI - Nursing scholarship bills raise few eyebrows. PMID- 12747309 TI - Healthcare Employee Protection Act. PMID- 12747310 TI - Tobacco settlement money for public health. PMID- 12747311 TI - Recognizing nursing's independent license: prescriptive authority for APNs. PMID- 12747312 TI - Environmental quality as a public health issue. PMID- 12747313 TI - Behind the curve? What we know and need to learn from public health systems research. PMID- 12747314 TI - A study of local public health system performance in Texas. AB - In summer 2001, 47 local public health agencies in Texas evaluated their public health systems using a modified version of the Local Public Health System Performance Assessment Instrument. A concurrent investigation sought to describe the performance of these systems in the aggregate and identify local health agency characteristics and other factors associated with system performance. High system performance was associated with larger community size, higher community socioeconomic status, greater public health agency capacity, and agency contribution to system performance. These findings raise questions concerning the expectation that all local public health systems, regardless of the core agency's capacity, should be able to attain the performance standards. PMID- 12747315 TI - Recommendations from testing of the National Public Health Performance Standards instruments. AB - The National Public Health performance Standards Program (NPHPSP) has developed assessment instruments based on the ten essential public health services (EPHS) for state and local health departments. The article reviews validity testing of the state and local instruments. The study employed multiple approaches to validity testing with state and local health departments in Florida, Hawaii, Minnesota, Mississippi, and New York. The New York State validity checks included the judgments of community partners. The study found that the EPHS have content and face validity as a basis for measuring public health system performance. The article includes recommendations for continued development of the NPHPS. PMID- 12747316 TI - A model for public health workforce development using the National Public Health Performance Standards Program. AB - Workforce development programs in public health should link improvements in workers' performance with improvements in their agencies' performance. The "ten essential services" of public health provide criteria for measuring both individual worker training (as in workforce competency standards) and agency performance (as in the Centers for Disease Control and Prevention's National Public Health Performance Standards Program). This shared foundation was the basis for a model strategic training program developed for use in a 500-employee urban county health department. Full implementation of this model as a foundation for assessment, curriculum development, and evaluation requires careful attention to management issues, confidentiality of employee records, and evaluation methodologies. PMID- 12747317 TI - Are the public health workforce competencies predictive of essential service performance? A test at a large metropolitan local health department. AB - Since many training initiatives employ the core public health workforce competencies as objectives, it is important to demonstrate an association between competency and essential service job performance. A cross-sectional survey of 420 employees of a local health department was conducted in 1999, with a response rate of 76 percent. Each of ten essential service performance measures was regressed on four core competency measures, controlling for employee experience and education. The competencies explained 2 percent to 20 percent of the variance in essential service performance. While offering support for the core competencies as a foundation for training program content, the results also make clear the large role that other individual, organizational, and community influences may have. Explaining additional variance in performance will require incorporating these variables into future studies. PMID- 12747318 TI - Are public health services available where they are most needed? An examination of local health department services. AB - Local health departments (LHDs) play a key role in the provision of public health services in the United States. Little is known about the extent to which LHD service availability varies by the socioeconomic characteristics of regional populations. This study merges data from the 1996 National Association of County and City Health Officials LHD profile survey and the Area Resource File system. The empirical analysis suggests that LHDs in low-socioeconomic background counties are more likely to provide services such as family planning. For other services, either LHD involvement is low across the board or the distribution of LHD services does not favor low-socioeconomic background counties. Thus, there is often room for improvements in service availability and targeting. PMID- 12747319 TI - Public health infrastructure system change: outcomes from the turning point initiative. AB - The Robert Wood Johnson and W.K. Kellogg Foundations created the Turning Point initiative to transform and strengthen the public health infrastructure. This study examined 135 public health system changes for their links to multiple sector collaborative engagement, essential public health services, health outcomes, and infrastructure building strategies. An on-line documentation system developed by the University of Kansas was used to record and analyze these linkages. The results showed that each state accomplished notable system changes; the majority involved more than one sector and primarily related to increasing local public health system capacity and organizational change. PMID- 12747320 TI - The Michigan Local Public Health Accreditation Program: many partners--one vision. AB - The Michigan Local Public Health Accreditation Program ensures the quality of local public health in Michigan by identifying and promoting the implementation of public health standards and evaluating and accrediting local departments' ability to meet them. The program is a collaborative effort between the Michigan Public Health Institute and the Michigan Departments of Agriculture, Community Health, and Environmental Quality. This article presents a case study. From the conceptual stage of designing a mode of collaboration to examining each step in the process, the goal is to provide a basic understanding of objectives of the accreditation program. The article concludes with the early results, providing a platform for future study. PMID- 12747321 TI - Using the National Neighborhood Indicators Partnership to improve public health. AB - The National Neighborhood Indicators Partnership (NNIP), a collaborative effort, uses local information in community building and policy making. A local intermediary in 19 NNIP partnership cities builds local data systems. Partners have learned five important lessons: (1) neighborhood-level data are essential for developing public policy, (2) technological advances have made it possible to maintain detailed local databases at relatively low cost, (3) various types of local organizations can serve as local partners, (4) good leadership is critical to building bridges across agencies, and (5) providing data is only the first step. Data must be used in ways that are visible, useful, and responsive to the community if the project is to succeed. PMID- 12747322 TI - A method to measure the impact of primary care programs targeted to reduce racial and ethnic disparities in health outcomes. AB - A retrospective population-based study was designed to test the impact on selected health outcomes of community-based primary care programs targeting racial and ethnic minorities. Zip codes were coded as either "high" or "low" access to targeted primary care programs to create the independent variable of interest. Outcome measures were chosen to represent unique dimensions of primary care. Generalized linear models were developed to compare rates for the outcome measures among blacks in high- and low-access areas. This study provides a useful approach that could be used to evaluate the impact of such programs in other communities. PMID- 12747323 TI - Community readiness to meet Healthy People 2010 targets. AB - Healthy People 2010 national objectives seek to engage public health systems in health improvement. Focusing on "healthy communities," the article describes the readiness of U.S. counties to meet targets and the technical assistance that may be needed. Using rate comparisons and trends, four readiness-to-improve health categories are calculated for nine mortality rates. Greatest readiness for improvement was found for coronary heart disease mortality, where the fewest jurisdictions (35%) were static, having neither met targets nor improved; least readiness (65%) was observed for colon and rectum cancer and suicide. Nevertheless, some counties (50-1,050) were exemplary, already meeting the target and still improving. PMID- 12747324 TI - Amanita muscaria: chemistry, biology, toxicology, and ethnomycology. AB - The fly agaric is a remarkable mushroom in many respects; these are its bearing, history, chemical components and the poisoning that it provokes when consumed. The 'pantherina' poisoning syndrome is characterized by central nervous system dysfunction. The main species responsible are Amanita muscaria and A. pantherina (Amanitaceae); however, some other species of the genus have been suspected for similar actions. Ibotenic acid and muscimol are the active components, and probably, some other substances detected in the latter species participate in the psychotropic effects. The use of the mushroom started in ancient times and is connected with mysticism. Current knowledge on the chemistry, toxicology, and biology relating to this mushroom is reviewed, together with distinctive features concerning this unique species. PMID- 12747325 TI - Double-stranded RNA elements associated with the MVX disease of Agaricus bisporus. AB - Double-stranded RNA (dsRNA) has been isolated from Agaricus bisporus fruit bodies exhibiting a wide range of disease symptoms. The symptoms which occurred singularly or in combination included; bare cropping areas on commercial beds (primordia disruption), crop delay, premature veil opening, off- or brown coloured mushrooms, sporophore malformations and loss of crop yield. All symptoms were associated with loss of yield and/or product quality. Collectively, these symptoms are described as mushroom virus X (MVX) disease. The dsRNA titre was much lower than that previously encountered with the La France viral disease of mushrooms and a modified cellulose CF11 protocol was used for their detection. A broad survey of cultivated mushrooms from the British industry identified dsRNA elements ranging between 640 bp and 20.2 kbp; the majority have not previously been described in A. bisporus. 26 dsRNA elements were identified with a maximum of 17, apparently non-encapsidated dsRNA elements, in any one sample. Three dsRNAs (16.2, 9.4 and 2.4 kbp) were routinely found in mushrooms asymptomatic for MVX. Previously, La France disease was effectively contained and controlled by minimising the on-farm production and spread of basidiospores. Our on-farm observations suggest that MVX could be spread by infected spores and/or mycelial fragments. PMID- 12747326 TI - Genetic structure of Fennoscandian populations of the threatened wood-decay fungus Fomitopsis rosea (Basidiomycota). AB - The genetic structure of five Fennoscandian populations of the threatened wood decay fungus Fomitopsis rosea (Basidiomycota) was investigated using codominant PCR-RFLP, allele specific amplification (ASA) markers, inter simple sequence repeat (ISSR) markers and mating studies. Sequence analyses of a subset of single spore isolates revealed sequence variation in four target sequences; internal transcribed spacer (ITS) and intergenic spacer (IGS1) of the nuclear ribosomal DNA, the translation elongation factor 1 alpha (efa) gene and the super oxide dismutase (sod) gene. No sequence variation was found in amplified portions of the mitochondrial large and small rRNA genes. Genotype distributions were mostly (90%) in accordance with Hardy-Weinberg expectations, and the nrDNA markers (ITS/IGS1), efa and sod were in most cases (87%) in linkage equilibrium, indicating an outcrossing reproductive mode, panmictic conditions and large population sizes of the fungus. Mating tests confirmed that F. rosea exhibits an outcrossing bipolar heterothallic mating system. Mating allele richness was high in two investigated populations. Phylogenetic analyses of ITS and IGS1 sequences from the five geographic populations revealed some geographic sub-structuring of the ITS sequences, but no sub-structuring of IGS1. The nrDNA (ITS/IGS1), efa and sod markers gave a low overall FST (0.013). The ISSR markers gave no clustering of the populations in UPGMA, and the between-population variance component was very low in AMOVA (0.4%), indicating a high level of gene flow. PMID- 12747327 TI - Phylogenetic relationship of Sclerotium rolfsii (teleomorph Athelia rolfsii) and S delphinii based on ITS sequences. AB - The phylogenetic relationships of the stem rot pathogens Sclerotium rolfsii and S. delphinii were examined, based on their rDNA ITS sequences. The ITS regions were cloned and sequenced to identify three distinct ITS types: r-1, r-2, and r 3. Two different ITS types exist within S. rolfsii and S. delphinii strains. Japanese strains and one strain of S. rolfsii from the USA contain types r-1 and r-2, whereas another strain from the USA and one from Chile have only one ITS type, r-2. S. delphinii strains have types r-1 and r-3. We discuss the implications of the common presence of ITS type r-1 for the taxonomy and evolution of this species complex. PMID- 12747328 TI - Shared ITS DNA substitutions in isolates of opposite mating type reveal a recombining history for three presumed asexual species in the filamentous ascomycete genus Alternaria. AB - About 15,000 species of ascomycete fungi lack a known sexual state. For fungi with asexual states in the anamorph genera Embellisia, Ulocladium, and Alternaria, six species have known sexual states but more than 50 species do not. In sexual filamentous ascomycetes, opposite mating type information at the MAT1 locus regulates mating and the opposite mating type genes each have a clonal, non recombining phylogenetic history. We used PCR to amplify and sequence fragments of the opposite mating type genes from three supposedly asexual species, A. brassicae, A. brassicicola and A. tenuissima. Each haploid fungal isolate had just one mating type, but both mating types were present in all the three species. We sequenced the ribosomal ITS regions for isolates of opposite mating type, for the three asexual species and four known related sexual species. In a phylogenetic analysis including other ITS sequences from GenBank, the three asexual species were not closely related to any of the known sexual species. Isolates of opposite mating type but the same species had identical ITS sequences. During any period of asexual evolutionary history, lineages of each mating type would have had a separate evolutionary history and any ITS substitutions shared between isolates of opposite mating type would have had to accumulate by convergence. Allowing for varying substitution rates and assuming a Poisson distribution of substitutions, the probability that isolates of opposite mating type shared an ITS substitution through convergence was low. This suggests that isolates of opposite mating type of A. brassicae, A. brassicicola and A. tenuissima were exchanging substitutions through sexual or parasexual reproduction while the ITS was evolving. If sexuality was lost, it was lost after the period of evolutionary history represented by the shared substitutions. PMID- 12747329 TI - Genetic diversity and virulence of Rhizoctonia species associated with plantings of Lotus corniculatus. AB - Species of Rhizoctonia cause a blight of Lotus corniculatus, a perennial forage legume. We characterized genetic variation and virulence in populations of R. solani and binucleate Rhizoctonia's associated with diseased L. corniculatus in field plantings over several years. Isolates of anastomosis groups AG-1 and AG-4 accounted for the R. solani recovered from diseased leaf and shoot tissues. Isolates of binucleate Rhizoctonia were recovered predominantly from soil and associated plant debris. Isolates of R. solani were more virulent on leaves and shoots of L. corniculatus than were binucleate Rhizoctonia isolates. Numerous unique DNA restriction patterns were observed among binucleate isolates and anastomosis groups of R. solani. Variation in restriction patterns was greater among isolates of AG-1 from the lower plant canopy than from the upper canopy. No restriction pattern was shared by any isolate from AG-1 and AG-4. Allelic and genotypic heterogeneity of AG-1 isolates were also greater in the lower plant canopy. Binucleate isolates exhibited greater heterogeneity than AG-1 isolates from either canopy region. L. corniculatus offers significant opportunities for investigating temporal and spatial dynamics of genetic structure of Rhizoctonia populations in perennial plant systems. PMID- 12747330 TI - Polymorphism of trichothecene biosynthesis genes in deoxynivalenol- and nivalenol producing Fusarium graminearum isolates. AB - Diversity in trichothecene mycotoxin production by 167 isolates of Fusarium graminearum was examined by chemical and molecular methods. Isolates from barley, corn, and wheat grown in Korea produced either deoxynivalenol (DON) or nivalenol (NIV), whereas isolates from corn grown in the United States produced DON only. Southern blotting of MseI-digested genomic DNA's from these isolates was performed using a 0.6-kb fragment of Tri5, a key enzyme for trichothecene production, as a probe. This technique revealed a single-band polymorphism between these isolates, with 1.8- and 2.2-kb bands arising from DON and NIV producers, respectively. The same set of isolates was subjected to previously developed PCR assays using primers derived from Tri7 or Tri13. These assays also revealed a single-band polymorphism between NIV- and DON-producing chemotypes. The polymorphisms at Tri5, Tri7, or Tri13 in all of the US isolates were consistent with their chemotypes as identified by GC-MS. However, for seven Korean isolates, chemical and molecular analyses yielded seemingly inconsistent results. This issue was resolved by Southern blot analysis with the Tri5 probe using two other restriction enzymes and sequence comparison of a 3.8-kb region spanning Tri5. In addition, one of these exceptional isolates was found to carry both DON and NIV chemotype-specific regions, possibly resulting from recombination between the two chemotypes. PMID- 12747331 TI - PCR-based DNA fingerprinting indicates host-related genetic variation in the nematophagous fungus Pochonia chlamydosporia. AB - The mitosporic fungus Pochonia chlamydosporia is a potential biocontrol agent for cyst (Heterodera spp. and Globodera spp.) and root knot (Meloidogyne spp.) nematodes, which are important agricultural plant pests. 54 isolates from diverse geographical regions and several nematode hosts were used in this study. Genetic variation was examined using enterobacterial repetitive intergenic consensus (ERIC) primed PCR and sequences from the internal transcribed spacer (ITS) rRNA region. ERIC PCR yielded 35 scorable binary characters from all the fungi tested and cluster analysis of the data showed that isolates from cyst nematodes were more genetically variable than those from root knot nematodes. The ITS regions were highly conserved, the only significant difference being an extra thymidine in isolates from Meloidogyne spp. Assays with nematode eggs indicated that isolates differ in their ability to infect different nematode genera. The results indicate host related variation in P. chlamydosporia. This finding has significant implications for the application of P. chlamydosporia as a biocontrol agent. PMID- 12747332 TI - Approaches for monitoring the release of Pochonia chlamydosporia var. catenulata, a biocontrol agent of root-knot nematodes. AB - Pochonia chlamydosporia var. catenulata is a potential biocontrol agent against root-knot nematodes. Diagnosis of isolates has relied on morphological identification, and is both time-consuming and difficult. beta-tubulin primers have been developed for the identification of this fungus that were specific enough to distinguish between varieties of the fungus within the same species. Separate primers have been developed for the specific detection of P. chlamydosporia var. catenulata based on ITS sequences, which were able to detect the fungus in soil from various sites in Cuba where the biocontrol agent had been added. When the PCR diagnosis was combined with serial dilution of soil samples on selective medium, colonies were rapidly identified. The fungus was still present, albeit at low densities, in soils inoculated five years previously. The development of a baiting method allowed quick in situ screening of the isolates' ability to infect nematode eggs, and when combined with PCR diagnosis both varieties of the fungus could be detected in infected eggs. RFLP analysis of ITS sequences from P. chlamydosporia provided an extra level of discrimination between isolates. PMID- 12747333 TI - Ascospore release and survival in Sclerotinia sclerotiorum. AB - The release and survival of ascospores of a UK Sclerotinia sclerotiorum isolate were studied. Apothecia placed in a spore clock apparatus with different lighting regimes at 15 degrees C released ascospores continuously with an increasing rate for the duration of experiments (72-84 h). Spore release was not confined to light or dark periods in alternating regimes and occurred in continuous dark or light. Ascospores were released in both saturated air (90-95% rh) and at 65-75% rh. High temperature and rh were detrimental to ascospore survival but spore viability was maintained for longer periods than previously reported. The significance of these results in relation to disease control is discussed. PMID- 12747334 TI - Morphology and physiology of the dimorphic fungus Mucor circinelloides (syn. M. racemosus) during anaerobic growth. AB - The dimorphic Mucor circinelloides requires an anaerobic atmosphere and the presence of 30% CO2 to grow as a multipolar budding yeast, otherwise hyphal growth predominates. Establishing other means to control the morphology would be a distinct advantage in the development of a fermentation process for this organism for the production of heterologous proteins. Thus, conditions suppressing polarised growth while at the same time abolishing the CO2 requirement were investigated in submerged cultivations. It was found that supplementing cultures with mixtures of ergosterol and Tween 80 resulted in yeast like growth under 100% N2. Their impact on growth and morphological development was assessed at a range of concentrations. Maximum biomass levels and the specific growth rate decreased at elevated levels of ergosterol and Tween 80. Possible effects of carbon dioxide and the added fatty acid/sterol mixture on supporting yeast growth by influencing the fluidity of the plasma membrane or affecting polarised growth are discussed. PMID- 12747335 TI - Production of wood-decay enzymes, mass loss and lignin solubilization in wood by tropical Xylariaceae. AB - Tropical xylariaceous taxa in the genera Biscognauxia, Hypoxylon and Xylaria were evaluated for their ability to produce wood-decay enzymes and effect mass loss and lignin solubilization in angiosperm and gymnosperm wood. All xylariaceous taxa were capable of cellulose and xylan hydrolysis, but few produced enzymes involved in lignin breakdown. Xylariaceous fungi were incapable of causing mass loss in gymnosperm wood, but several caused significant mass loss in angiosperm wood during a six month in vitro exposure. Mass loss values obtained were low at approx. 20% of those obtained for basidiomycetes. Lignin was solubilized at similar rates to mass loss by Hypoxylon and Xylaria species, resulting in indices of lignin solubilization between 0.88 and 1.11, which were similar to those obtained for white-rot basidiomycetes and higher than those previously reported for any other xylariaceous fungi. PMID- 12747336 TI - New species, notes and key to the aeroaquatic genera Beverwykella and Ramicephala gen. nov. AB - Two new aeroaquatic fungi, Ramicephala sphaerospora gen. sp. nov. and Beverwykella clathrata sp. nov., are described and illustrated from tropical and subtropical latitudes. B. clathrata, a species similar to B. cerebriformis, deviates from the latter in having a clathrate outermost cell layer instead of tightly appressed cells, a feature unique in Beverwykella. The second species, Ramicephala sphaerospora gen. sp. nov., has a unique combination of features concerning conidial ontogeny and morphology, and is therefore placed in a new genus. The genus most similar to Ramicephala is Beverwykella, from which it differs in having: (1) globose, non-flattened conidia; (2) a large central globose cell giving rise to centrifugally branching, tightly packed chains of cells; and (3) a compact peripheral cell layer, the cells of which are tightly interlocked in a zipper-like form. Similarities of R. sphaerospora to species of Pseudaegerita, Candelabrum and Cristulariella are discussed. Illustrations of and a key to all described species of Ramicephala and Beverwykella are provided. PMID- 12747337 TI - First report of Basidiolum fimbriatum since 1861, with comments on its development, occurrence, distribution and relationship with other fungi. AB - An obscure parasitic fungus, Basidiolum fimbriatum, was found on Amoebidium parasiticum (Amoebidiales) associated with Caenis sp. (mayfly) nymphs, during a survey of gut fungi (Trichomycetes) from a small stream in northeastern Kansas, USA. The hindguts of the nymphs harboured a species of Legeriomycetaceae and Paramoebidium sp. This is the first report of the ectocommensal protozoan, A. parasiticum, associated with the gills of Caenidae (Ephemeroptera), and of B. fimbriatum in the 142 years since its original documentation from Wiesbaden, Germany. B. fimbriatum is recorded from two midwestern USA states (Kansas and Iowa) and the morphological and developmental features of the parasite on its host are compared with Cienkowski's original observations and interpretation. B. fimbriatum is characterized as a parasitic fungus possessing merosporangia that from on a simple pyriform thallus that penetrates and consumes its host via a haustorial network. The hypothesis that B. fimbriatum is most closely related to members of the order Zoopagales sensu Benjamin (1979) is proposed. The importance of future collections and molecular-based phylogenetic approaches to place this parasitic fungus within a current system of classification are highlighted. PMID- 12747339 TI - Volume-dependent or pressure-dependent effect on renal function after spinal anesthesia. PMID- 12747338 TI - Sooty moulds from European tertiary amber, with notes on the systematic position of Rosaria ('Cyanobacteria'). AB - Sooty moulds are described and illustrated from European amber dating back to 22 54 Myr. All the fossils are fragments of superficial subicula composed of brown moniliform hyphae with markedly tapering distal ends. The subglobose cells are identical to those of extant Metacapnodium (Metacapnodiaceae, Capnodiales) species. Also other preserved features, like the type of apical growth, wide angled branching and the production of two distinctive conidial states, supports a placement in this genus. The fossils demonstrate that Metacapnodium hyphae have remained unchanged for tens of millions of years. This confirms that hyphal morphology and conidial states should be accorded considerable classificatory significance in this group of fungi. The following nomenclatural change is made: Metacapnodium succinum comb. nov. (syn. Rosaria succina). The type specimen was initially described as a filamentous cyanobacterium, due to similarities with Rosaria ramosa. Also the systematic position of this attribute is shortly discussed. PMID- 12747340 TI - A randomized, prospective comparison of end-tidal CO2 pressure during laparoscopic cholecystectomy in low and high flow anesthetic system. AB - BACKGROUND: Low flow anaesthesia has been used in anesthetic practice to prevent operation room pollution and also for econormical reasons. Since the safety of low flow technique has not been clearly determined in previous researches this study was performed to compare the end-tidal CO2 pressure during laparoscopic cholecystectomy in low flow as opposed to high flow anesthesia. METHODS: Forty patients, 30-65 years of age, ASA physical status I or II, scheduled for laparoscopic cholecystectomy under general anesthesia were randomly alloted to low flow and high flow groups. End-tidal CO2 pressure, arterial blood pressure, pulse oximetric oxygen saturation, and heart rate were measured before, during and after insufflation of CO2 into the peritoneal cavity. The data were compared between two groups. RESULTS: Pneumoperitoneum caused a decrease in PaO2 and oxygen saturation together with increases in PaCO2 and end-tidal CO2 pressure in both groups but the differences between two groups were not statistically significant. CONCLUSIONS: The result of this study shows that end-tidal CO2 pressure during laparoscopic cholecystectomy using low flow anesthesia system is comparable to that with high flow system. Therefore it can be concluded that low flow anesthesia can be used with relative safety in anesthetic management of patients during laparoscopic cholecystectoy. PMID- 12747341 TI - Fluid administration prevents renal dysfunction during hypotension under spinal anesthesia in a rat model. AB - BACKGROUND: Severe hypotension deteriorates renal functions and renal hemodynamics especially renal cortical blood flow. Systemic hypotension following high level spinal anesthesia may impair renal functions in spite of the blockade of renal sympathetic nerves that may help prevent vasoconstriction. Fluid loading is clinically applied for preventing hypotension but the effects on the changes of renal functions have not been studied. This study was designed to investigate the effects of fluid loading on systemic hemodynamics, renal hemodynamics and functions especially the blood distribution to renal cortex. METHODS: A rat model was used in our study. Intravenous normal saline infusion was started in both control group (5 ml/kg/h, 8 rats) and fluid loading group (15 ml/kg/h, 8 rats) 30 min before spinal anesthesia. A high level (above T4) spinal anesthesia was conducted via a preset intrathecal catheter with 0.5% hyperbaric bupivacaine. Blood pressure, heart rate and renal cortical microvascular blood flow (CMBF) were measured via a laser Doppler probe firmly contacted on renal cortex and recorded continuously after spinal anesthesia. Renal functions including glomerular filtration rates (GFR, by inulin clearance), effective renal plasma flow (ERPF, by P-aminohippurate clearance), urine flow rate (UFR) and electrolytes excretion were measured every 30 min after spinal anesthesia. RESULTS: Severe hypotension was notable within 5-10 min after intrathecal anesthesia and recovered with 30 min in both groups but the difference was not significant between groups. In the control group, GFR and ERPF decreased significantly in the first 30 min by 51.9 +/- 19.8% and 44.3 +/- 13.7% respectively (P < 0.05) and recovered after 60 min. Also the deteriorations of UFR and CMBF were significantly longer (over 60 min). In fluid loading group, ERPF, UFR and CMBF could maintain throughout the experiment but only GFR was affected in the first 30 min. CONCLUSIONS: Fluid administration did not prevent hypotension following high level spinal anesthesia but might have beneficial effects on renal hemodynamics especially on the renal cortical circulation and urine flow rate. PMID- 12747342 TI - Performance evaluation of quality improvement team in an anesthesiology department. AB - BACKGROUND: In health care community, quality improvement pathway has always been treated as critical index to control cost, improve efficiency and promote service quality, particularly in the last decade. From theoretical standpoint, clinical practice as well as research data, quality improvement team has been demonstrated to play an important role in the adaptation to the changing health environment and enhancement of the competition through the improvement process. The purpose of this study was to explore members' job satisfaction, morale, organizational commitment and inventory management through quality improvement team intervention in a department of anesthesiology. METHODS: This study was of a quasi experimental and longitudinal design. The subjects involved 45 nurse anesthetists (the experiment group, intervention of quality improvement team) and 50 operation room nurses (control group) in a general hospital. The quality improvement team had been initiated and implemented pursuant to the quality improvement process for 8 months. GEEs (Generalized Estimating Equations) model was used to examine the differences in job satisfaction, morale, organizational commitment, and the inventory management was also examined between two groups. RESULTS: After control of all variables, except education background, such as age, marital status, education, position and nursing experience, a natural growth effect was observed on quality improvement team. The results revealed that the experimental group showed significant positive effects on both job satisfaction and organizational commitment after the intervention. The morale scale did not differ significantly between two groups. In the inventory management, the experimental group successfully decreased the monthly consumable materials stock with a descending rate of 24.8%, while in the control group, the inventory was increased 16.9% in the basal stock instead. CONCLUSIONS: As other previous reports did, the present study also demonstrated that intervention of the quality improvement team improves the nurse anesthetists' job satisfaction, such as promotion of autonomy, organizational policy and positive member interaction. Moreover, it improves work efficiency, service quality as well as control of the stock inventory. PMID- 12747343 TI - Epidural ropivacaine for postoperative analgesia in Taiwanese patients. AB - BACKGROUND: Ropivacaine is the latest long-acting amide local anesthetic. As it is less cardiovasculotoxic and neurotoxic than bupivacaine it is an attractive anesthetic agent used in clinical anesthesia and postoperative analgesia. This study was undertaken to seek for a suitable dosage of ropivacaine in postoperative analgesia for Taiwanese patients whose average physicality is not entirely compatible with the pharmacopeially recommended dosage for western people. METHODS: For assessment of epidural ropivacaine for postoperative analgesia 105 adult patients were enrolled and randomly allotted to three groups. Patients in Group A were given epidurally 0.15% ropivacaine, while those in Group B and Group C were given 0.125% and 0.10% ropivacaine respectively. Pain was evaluated with visual analogue scale (VAS) and modified Bromage scale, and adverse effects were recorded at the designated points of time during the postoperative 24-hour period. RESULTS: The demographic profiles were comparable among three groups. In VAS score, Group A (3.20 +/- 0.47) and B (3.11 +/- 0.41) did not differ much, while Group C (3.97 +/- 0.71) the score was signally higher than Group A and Group B (P < 0.05). Adverse effects, such as paraesthesia, nausea and urinary retention were observed more in Group A. CONCLUSIONS: From the results of this study, we are of the opinion that 0.125% ropivacaine could provide a postoperative analgesia in Taiwanese patients to their satisfaction with less adverse effects. PMID- 12747344 TI - The analgesic effect of oral morphine or pentazocine for extracorporeal shock wave lithotripsy. AB - BACKGROUND: Extracorporeal shock wave lithotripsy (ESWL) in these days is usually carried out on ambulatory or outpatient basis. With the application of a lithotriptor of modern version an appropriate yet cost-effective analgesia with minimal side effects for ESWL is mandatory. METHODS: The analgesic effect of oral morphine (30 mg) was compared with that of pentazocine (100 mg) in a prospective study comprising 100 patients undergoing ESWL with a lithotripter of improved version for urinary tract stones. All patients received orally lorazepam 1 mg as sedative together with the appointed tested drug 30 min before the procedure. The analgesic effects of both drugs were assessed having recourse to the pain scale and efficacy scale. RESULTS: There were 94% of patients in the pentazocine (mixed agonist-antagonist) group who felt satisfied with the regimen and stood the procedure well without resort to supplemental drug, as compared with the morphine (potent mu-agonist) group in which only 70% of patients did so. Although the adverse effect such as dizziness was found in the pentazocine group, the degree of sleepiness produced by its deeper sedation effect was to the advantage of patients during the lithotripsy procedure. There were no significant changes in intergroup mean blood pressure (MBP), but heart rate (HR) was higher and O2 saturation (SpO2) was lower in the pentazocine group after treatment. Both narcotics did not induce renal colic in our study. Also, pentazocine 100 mg plus lorazepam 1 mg given orally did not induce psychotomimetic reaction intraoperatively or postoperatively. CONCLUSIONS: We concluded that oral pentazocine at 100 mg plus lorazepam 1 mg, could offer satisfactory analgesia in patients undergoing ESWL for urinary tract stones with a lithotripter of improved version. PMID- 12747345 TI - Esophageal perforation after tracheal intubation, spontaneous or iatrogenic?--a case report. AB - Esophageal perforation is a rare but life-threatening complication associated with tracheal intubation, especially after difficult intubation. Esophageal perforation after anesthesia is rare and usually secondary to esophageal instrumentation. Spontaneous esophageal perforation following forceful vomiting (Boerhaave's syndrome) is also extremely rare and has some risk factors. We present a case of perforation of esophagus after cataract surgery under general anesthesia with gentle orotracheal intubation and discuss the possible mechanisms responsible for this unusual disease entity. The patient underwent successful surgical repair and was still alive 4 years after the operation. PMID- 12747346 TI - Accidental subdural catheterization due to complication of epidural anesthesia--a case report. AB - Although accidental subdural injection is a well-recognized complication of epidural block, only a mere handful cases have been substantially proven by radiological evidence. Here we report a case of subdural catheterization during the attempt of epidural anesthesia for a gynecological procedure. Its clinical course and radiological findings are compared with those of the cases previously reported in literature. Whenever there is the occurrence of widespread of sensory block together with respiratory distress and hemodynamic unstability following epidural injection of local anesthetic, a subdural injection should be considered in spite of a negative confirmation. Repeated subdural injection of a local anesthetic at the same site may predispose patients to serious morbidity. Therefore, we recommend that when a subdural injection is evident or suspected, reinsertion of the catheter in the epidural space via another entry or contemplation of a switch to another anesthetic technique is mandatory. PMID- 12747347 TI - Postoperative pulmonary edema, transfusion-related?--a case report. AB - Transfusion-related acute lung injury (TRALI) is a severe reaction between leukocyte antigen and antibody during transfusion of plasma-containing components. Recently, biologically active lipids have been also suggested to cause the disorder. It is a rare, but rather benign pulmonary edema. We report a postoperative pulmonary edema, which was temporally and clinically compatible with TRALI. Because the patient received blood products from 3 or 4 donors and the disorder was not recognized right away, the laboratory task for the definite diagnosis was difficult. Nevertheless, the patient had fully recovered in 36 hours after supportive therapies. Without identifying the blood donor implicated in the disorder, transfusion reactions or TRALI will be inevitable. PMID- 12747348 TI - Delayed hypoxemia after bone cement insertion during total hip replacement under spinal anesthesia--a case report. AB - We report a case of delayed hypoxemia in an aged healthy male patient, which developed 2 hours after cementation of the prosthesis in total hip replacement (THR) under spinal anesthesia. The patient was doing well throughout the operation but unfortunately, progressive tachypnea was noted 1 h after he was transferred to the recovery room (i.e. 2 h after the application of bone cement into the femur). An hour further, distinct wheeze was heard bilaterally on auscultation, which signified bronchospasm. Arterial blood gases analysis revealed a low PaO2 of 71 mmHg and a decrease of oxygen saturation to 91% with supplement of fractional oxygen of 35%. Aerosolization of bronchodilator with terbutaline was administered and supplemental fractional oxygen was increased to 50%. Although wheezing soon subsided, tachypnea and desaturation persisted. He was then transferred to the surgical intensive care unit for further management. Ventilation-perfusion lung scan was performed, which was suggestive of multiple pulmonary embolism. PMID- 12747349 TI - Inquiring into indicators and origin of catastrophic events at stratigraphic boundaries. AB - Since 1982, numerous indicators of catastrophic events have been observed at the main stratigraphic boundaries in China during the Phanerozoic, i.e. Precambrian Cambrian, Permian-Triassic, Cretaceous-Tertiary as well as Ordovician-Silurian and Devonian-Carboniferous boundaries. These markers are boundary clay layer, microspherules, high Ir anomaly, mass extinction of organisms and impact of meteorite. We support the hypothesis of the extraterrestrial origin of catastrophic events and suggest that the events at different stratigraphic boundaries might be different in features and processes. PMID- 12747350 TI - Global summit of national bioethics commissions: Tokyo communique (1999). PMID- 12747351 TI - Mental disorders and genetics: the ethical context: conclusions and recommendations. PMID- 12747352 TI - IVF-related research (executive summary). PMID- 12747353 TI - Composition of the International Bioethics Committee of UNESCO (IBC). PMID- 12747354 TI - Research involving persons with mental disorders that may affect decisionmaking capacity. AB - In December 1998, the US National Bioethics Advisory Commission (NBAC) published the above-named report. The committee made twenty-one recommendations, some of which provide a set of requirements that NBAC believes should be satisfied in all research involving persons with mental disorders, while some are optional protections, that may be considered appropriate, depending on the particular circumstances of the research. The full recommendations are reprinted below. PMID- 12747356 TI - [Transfusion of erythrocyte homologues: products, indications, alternatives]. PMID- 12747357 TI - On euthanasia: blindspots in the argument from mercy. AB - In the euthanasia debate, the argument from mercy holds that if someone is in unbearable pain and is hopelessly ill or injured, then mercy dictates that inflicting death may be morally justified. One common way of setting the stage for the argument from mercy is to draw parallels between human and animal suffering, and to suggest that insofar as we are prepared to relieve an animal's suffering by putting it out of its misery we should likewise be prepared to offer the same relief to human beings. In this paper, I will argue that the use of parallels between human and animal suffering in the argument from mercy relies upon truncated views of how the concept of a human being enters our moral thought and responsiveness. In particular, the focus on the nature and extent of the empirical similarities between human beings and animals obscures the significance for our moral lives of the kind of human fellowship which is not reducible to the shared possession of empirical capacities. I will suggest that although a critical examination of the blindspots in these arguments does not license the conclusion that euthanasia for mercy's sake is never morally permissible, it does limit the power of arguments such as those provided by Rachels and Singer to justify it. I will further suggest that examination of these blindspots helps to deepen our understanding of what is at stake in the question of euthanasia in ways that tend otherwise to remain obscured. PMID- 12747358 TI - Just how unlawful is "euthanasia"? AB - Those who campaign for law reform to permit "euthanasia" may seek different things and at least some of what they seek may already be permissible under the criminal law of England and Wales. In this paper I examine one means whereby the criminal law delivers outcomes acceptable to the euthanasia lobby, that is the curious notion of "causation" deployed by the law, which adds a value override to the more usual notion of factual causation such that, for example, if medical treatment falls within the acceptable range as normal and proper, the pre existing injury or illness is treated as exclusively the cause of death and the doctor escapes criminal liability, even where the medical treatment will shorten life to the certain knowledge, possibly even the wish, of the doctor. Thus the law may already be delivering a range of outcomes--euthanasia in a weak sense- acceptable to the euthanasia lobby. If so, it achieves this by stealth. That is inappropriate to the doctor-patient relationship, which is one of trust. So there is a strong case for greater transparency. Moreover, there are limits to the acceptable outcomes which an unreformed criminal law can deliver and in a range of cases the criminal law condemns the doctor to impotence and the patient to a prolonged, miserable and undignified death. So there is also a case for going beyond the current law and legalising euthanasia in a strong sense. PMID- 12747359 TI - Suicide intervention and non-ideal Kantian theory. AB - Philosophical discussions of the morality of suicide have tended to focus on its justifiability from an agent's point of view rather than on the justifiability of attempts by others to intervene so as to preserve it. This paper addresses questions of suicide intervention within a broadly Kantian perspective. In such a perspective, a chief task is to determine the motives underlying most suicidal behaviour. Kant wrongly characterizes this motive as one of self-love or the pursuit of happiness. Psychiatric and scientifc evidence suggests that suicide is instead motivated by nihilistic disenchantment with the possibility of happiness which, at its apex, results in the loss of the individual's conception of her practical identity. Because of this, methods of intervention that appeal to agents' happiness, while morally benign, will prove ineffective in forestalling suicide. At the same time, more aggressive methods violate the Kantian concern for autonomy. This apparent dilemma can be resolved by seeing suicide intervention as an action undertaken in non-ideal circumstances, where otherwise unjustified manipulation, coercion, or paternalism are morally permitted. PMID- 12747361 TI - Communication in the development of culturally competent palliative care services in the UK. PMID- 12747360 TI - Autonomy, constraining options, and organ sales. AB - Although there continues to be a chronic shortage of transplant organs the suggestion that we should try to alleviate it through allowing a current market in them continues to be morally condemned, usually on the grounds that such a market would undermine the autonomy of those who would participate in it as vendors. Against this objection Gerald Dworkin has argued that such markets would enhance the autonomy of the vendors through providing them with more options, thus enabling them to exercise a greater degree of control over their bodies. Paul Hughes and T.L. Zutlevics have recently criticized Dworkin's argument, arguing that the option to sell an organ is unusual in that it is an autonomy undermining "constraining option" whose presence in a person's choice set is likely to undermine her autonomy rather than enhance it. I argue that although Hughes' and Zutlevics' arguments are both innovative and persuasive they are seriously flawed--and that allowing a market in human organs is more likely to enhance vendor autonomy than diminish it. Thus, given that autonomy is the preeminent value in contemporary medical ethics this provides a strong prima facie case for recognizing the moral legitimacy of such markets. PMID- 12747362 TI - Professional misconduct. PMID- 12747363 TI - Cosmetic surgeon--discipline. PMID- 12747364 TI - Effect of gravitational sedimentation on simulated aerosol dispersion in the human acinus. AB - We studied the effect of gravitational sedimentation on the dispersion of 0.5 and 1 micrometer-diameter particle boluses within a two-dimensional symmetric six generation model of the human acinus. Boluses were introduced at the beginning of a 2-s inspiration immediately followed by a 4-s expiration, in normal gravity (1 G) and in the absence of gravity (0 G). The flow corresponded to a flow rate at the mouth of 500 ml/s. In 0 G, simulated dispersion (Hsim) was 16 ml for both particle sizes. In 1 G, Hsim was 71 and 242 ml for 0.5 and 1 micrometer-diameter particles, respectively, showing the effect of gravitational sedimentation. The difference between experimental data (J. Appl. Physiol. 86 (1999) 1402) and simulations was independent of particle size. This suggests that the residual dispersion was independent of the intrinsic properties of the particles and was more likely due to other mechanisms such as ventilation inhomogeneities, cardiogenic oscillations and alveolar wall motion. PMID- 12747365 TI - Outpatient cardiology care improves survival odds after heart attack. PMID- 12747366 TI - ICU care management by intensivists reduces mortality and length of stay. PMID- 12747367 TI - Not-for-profit dialysis center care carries lower mortality risk, study says. PMID- 12747368 TI - Having pharmacists participate in medical rounds reduces medication errors. PMID- 12747370 TI - HIPAA Possumus. PMID- 12747369 TI - Exposure of normal and transformed cells to nevirapine, a reverse transcriptase inhibitor, reduces cell growth and promotes differentiation. AB - Endogenous, nontelomeric reverse transcriptase (RT) is encoded by two classes of repeated elements: retrotransposons and endogenous retroviruses. Expression of RT coding genes is generally repressed in differentiated nonpathological tissues, yet is active in the mammalian germ line, embryonic tissues and tumor cells. Nevirapine is a non-nucleoside RT inhibitor with a well-characterized inhibitory activity on RT enzymes of retroviral origin. Here, we show that nevirapine is also an effective inhibitor of the endogenous RT in murine and human cell lines. In addition, progenitor and transformed cells undergo a significant reduction in the rate of cell growth upon exposure to nevirapine. This is accompanied by the onset of differentiation, as depicted in F9 and C2C7 progenitor cells cultures in which nevirapine triggers the expression of differentiation-specific markers. Consistent with this, an extensive reprogramming of cell cycle gene expression was depicted in nevirapine-treated F9 cultures. Furthermore, nevirapine exposure rescued the differentiation block present in acute myeloid leukemia (AML) cell lines and primary blasts from two AML patients, as indicated by morphological, functional and immunophenotypic assays. The finding that an RT inhibitor can modulate cell proliferation and differentiation suggests that RT may represent a novel target in the development of therapeutical approaches to neoplasia. PMID- 12747371 TI - Thalidomide administration for the treatment of resistant plasma cell leukemia. PMID- 12747372 TI - Successful treatment of invasive pulmonary aspergillosis by transbronchial injection of urinary trypsin inhibitor and amphotericin B. PMID- 12747374 TI - Research into RIK and Nimbus 3 mattresses. PMID- 12747375 TI - [Re: W.F. Haupt, W. Holling: Diagnosis of brain death: medical and legal aspects under the transplantation law of the German Republic. Fortschr Neurol Psychiat 2002; 70: 583-590]. PMID- 12747373 TI - Effects of recombinant factor VIIa in haemorrhagic complications of urological operations. PMID- 12747376 TI - [Re: W.F. Haupt, W. Holling: Diagnosis of brain death: medical and legal aspects under the transplantation law of the German Republic Fortschr Neurol Psychiat 2002; 70: 583-590]. PMID- 12747377 TI - [Moral harassment at work. (Law no. 2002-73 of January 17, 2002; law no. 2003-6 of January 3, 2003)]. PMID- 12747378 TI - Chronic digestive bleeding in blue rubber-bleb nevus (Bean syndrome). PMID- 12747379 TI - Err or just air? Free intraabdominal air: not always a reason for surgery. PMID- 12747380 TI - [Psychiatric therapy and pharmacology in medieval Islam]. AB - Although psychiatric therapy and pharmacology in Medieval Islam are based on the ancient Greek tradition, the original Arabic contribution in the introduction and employment of new substances is undeniable. Another important aspect which received a decisive impetus by Arab physicians was the concept of psychical therapy. PMID- 12747381 TI - Mechanism of injury is important. PMID- 12747382 TI - Capnography lessons. PMID- 12747383 TI - Whatever happened to "child and adolescent psychiatry"? PMID- 12747386 TI - Tourette syndrome and prognosis in autism. PMID- 12747387 TI - [The origins of the Islamic model of hospital]. AB - In this paper a new perspective is proposed regarding the most relevant factors in the origin and the first developments of the Islamic hospital in Abbasid Baghdad. Notably, the importance of the Persian contribution to the foundation of the first hospitals is questioned and the major focus is put on the role of Eastern Christian assistential institutions. The rise of the Islamic hospital is reconsidered in the frame of the wider process of urbanisation in the first Abbasid century. The Islamic bimaristan brought about a new concept of assistance by offering a medical cure rather than care. PMID- 12747388 TI - Drug use and the role of homelessness in the process of marginalization. AB - The marginalization theory of life histories implies that drug users who are considered as marginalized manifest more serious social, economical, physical, and psychological problems than nonmarginalized drug users. The degree of marginalization is assumed to be an indicator of homelessness. The theory argues that homelessness is a stage in the life of a user that is associated with the loss of control of use. In this paper the effects of the dimensions of marginalization on homelessness are reported. The marginalization theory emerged from ethnographic fieldwork research among the drug users population in Parkstad Limburg, the Netherlands. Ethnographic fieldwork is often restricted to a (selective observed) part of the total population. To verify whether the marginalization theory was valid for the total unknown population, we used quantitative data obtained in 1999 by a two-mode network sample (n = 58). As a conclusion homelessness was more likely to be present among marginalized than nonmarginalized drug users. PMID- 12747389 TI - Homeless drug users in Rotterdam, the Netherlands: profile, way of life, and the need for assistance. AB - Decreasing the number of homeless drug users is one of the main characteristics of inner city drugs policy. The present study selected an urban-ethnographic perspective (the subculture theory) in order to explore why one drug user is homeless and another 7 not, and to attempt to describe and define the homeless and their immediate social environment. These issues were formulated into the following research questions: 1. What are the sociodemographic characteristics of homeless drug users in Rotterdam, and do they differ from domiciled drug users? 2. What are their living conditions? 3. What are the reasons for being homeless? 4. Does the period of homelessness play a role in the need to change one's lifestyle? Five research methods were employed for this study: a literature search, interviews with key persons, field notes from community fieldworkers, a survey among drug users (n = 204), and photographic reports from six homeless users. Data were collected in 1998/1999. The results document that in our study population there were more women, more illegal persons, and more foreigners than among domiciled drug users, and that the homeless group used heroin and cocaine on more days. A large proportion of the homeless users had no identity papers and no health insurance. This did not, however, lead to more self-reported sickness or a higher prevalence of infectious diseases compared with nonhomeless drug users. Easily accessible (low threshold) social care centers and assistance are very important. Few of the homeless had voluntarily chosen a homeless life-most describe an event that was a trigger for their homelessness. The average duration of being homeless was 17 months, and the longer someone had been homeless the less inclined they were to change their situation. This paper also discusses policymaking implications. PMID- 12747390 TI - Unmet drug and alcohol service needs of homeless people in London: a complex issue. AB - Little research has been conducted on the drug use of those who sleep rough (on the streets) in the United Kingdom (UK). During 2000, to fill in the gaps in the knowledge base, researchers at the National Addiction Centre, London, carried out a community survey using a structured questionnaire amongst 389 homeless people recently or currently sleeping rough, in order to investigate their met and unmet drug and alcohol service needs. In total, 265 (68%) had a need for drug services and 97 (25%) for alcohol services. Over half of the current drug users (170/324, 52%) and 88 (33%) of the 264 current alcohol users wanted help with their substance use, but few were currently accessing the appropriate services, other than needle exchanges. The challenge for services is to build these potential clients' motivation to accept health-conferring intervention. PMID- 12747391 TI - Course of alcoholism in homeless men in Munich, Germany: results from a prospective longitudinal study based on a representative sample. AB - In an earlier paper (Fichter, M. M., Quadflieg, N. (1999). Alcoholism in homeless men in the mid-nineties: results from the Bavarian Public Health Study on Homelessness. European Archives of Psychiatry and Clinical Neuroscience 249:34 44), we reported data on alcoholism and comorbidity in 265 homeless men in Munich. There-as in this paper-we divided the sample into three groups based on a lifetime diagnosis of alcohol dependence (N = 187), alcohol abuse (N = 17), and no diagnosis of alcoholism (N = 61) at baseline assessment. This study reports a three-year prospective longitudinal assessment of the original representative sample of homeless men in Munich. Interviews at baseline and at follow-up included the SCID-I and covered several other areas (cognitive impairment, somatic complaints, use of medical services, and other psychosocial variables). Of 247 homeless men still alive, at three-year follow-up, 185 (74.9%) were successfully traced and personally interviewed. Alcohol dependency in homeless men at first wave assessment (as compared to men not manifesting alcohol abuse or dependence) was associated with a higher proportion of homelessness at three-year follow-up, an increase of alcohol consumption at three-year follow-up, reduction of monthly income, higher death rate, and high use of general medical services but very low utilization of (specific substance) user treatment services. Alcoholism in homeless men constitutes a posited risk factor for an unfavorable course over time with regard to such a person's living situation and health status in spite of more utilization of medical services. PMID- 12747392 TI - Homelessness and high-dosage buprenorphine misuse. AB - AIM: To determine whether intravenous drug users (IDUs) are more likely to misuse high dosage buprenorphine (HDB) if they are homeless. DESIGN: We carried out a cross-sectional study using data collected from HDB users between 1998 and 1999. Data were collected by use of a structured questionnaire with questions about demographic characteristics, and use of HDB and other substances. IDUs were considered to be homeless if they did not live on their own or with their parents or friends. SETTING: IDUs were recruited from three centers for the treatment of drug users, three health care networks, one prison, one sleep-in, and two centers that provide psychosocial support for IDUs. PARTICIPANTS: Of the 788 eligible patients, 779 answered the questionnaire (response rate: 98.9%). RESULTS: Homeless IDUs were more likely to have injected HDB than those who were not homeless (67% vs. 47%; p<0.001), and their injection behaviors were more likely to be unsafe. The first HDB injection was more likely to result in medical complications in the homeless group than in the nonhomeless group (58% vs. 38%; p=0.001). Homeless IDUs were less likely to receive medical followed-up and were less well informed about the correct way of using HDB than nonhomeless IDUs. CONCLUSION: Homeless IDUs are more likely to misuse HDB. Thus, HDB maintenance therapy may not be the most appropriate maintenance therapy for this group. PMID- 12747393 TI - Estimating the use of illegal drugs among homeless people using shelters in Denmark. AB - Based on a longitudinal study of approximately 1000 homeless people who used shelters or similar institutions in 1988-89, an estimate was made of the rate of people using illegal drugs in that population. This estimation was made possible by combining information from different registers-criminal records, enrollments at drug user treatment centers, and registers indicating cause of death. The samples use of drugs is also noted. The implications of these findings are discussed in the light of Denmark's current policy towards people using illegal drugs and using shelters. PMID- 12747394 TI - Alcohol and other drug use disorders among homeless people in Australia. AB - This paper describes alcohol and drug-use disorders among 210 homeless people in Australia, and compares the Australian findings with the international literature. While the prevalence of alcohol-use disorders among people who are homeless in Australia is comparable with other international studies, drug-use disorders appear to be more prevalent among Australian homeless than comparable international studies. Reasons for this difference are explored. PMID- 12747395 TI - Deviance over the life course: the case of homeless substance abusers. AB - Characteristics of the homeless population highlight current and emerging social problems in America. It is estimated there are two to three million homeless people in the United States and that between 20% and 80% have substance abuse problems. In this paper I apply Hirschi's social bond theory to a qualitative study of 31 male homeless substance users from New Haven, Connecticut. I interviewed each man once in 1992 and a second time during 1993. The interview data provide evidence of non-normative attachments to families, friends, and institutions where there is little commitment to conventional goals. I conclude that for these men, being homeless and abusing substances is normative to their lives. PMID- 12747396 TI - The individual and beyond: a socio-rational choice model of service participation among homeless adults with substance abuse problems. AB - While substance user service programs can help homeless adults solve their substance use and housing problems, relatively few needy individuals use and complete these programs. The lack of participation is poorly explained by typical empirical studies, most of which consider the role in service participation of various personal traits and client problems. The current article instead seeks to explain service participation through the application of an alternative, "socio rational choice" model. This model has three premises: Clients weigh the costs and benefits of participating in services against alternative uses of their time and resources. The clients' weighing procedures reflect their personal situations and perceptions of the treatment environment. The perceptions of their personal situations and perceptions of the treatment environment are affected by the manner in which clients react to representatives of service systems, members of their social network including both housed and homeless persons, and other individuals. Secondary evidence supports many of the model's hypotheses and generally suggests that homeless clients may be heavily affected by their experiences with individuals and systems with which they come into contact. PMID- 12747397 TI - Homelessness and substance misuse: a tale of two cities. AB - In this article we examine the relationship between alcohol and drug misuse among the literally homeless (those living out of doors and in emergency shelters) in Hartford, Connecticut and Providence, Rhode Island, two northeastern U.S. cities of comparable size. We worked with homeless advocacy organizations in both cities, using a point-in-time census (N = 1058) and random sample (N = 66) in Hartford, and a sampling of clients (N = 82) of six shelters serving residents of Providence (N = 82). We found substance misuse relevant in 47.2% of the homeless in Hartford and in 45.1% of the homeless in Providence. We conclude that there is a great need for substance treatment services inside shelters, soup kitchens, and day centers so that homeless individuals have an opportunity to engage in treatment within their own milieu. PMID- 12747398 TI - Disaffiliation, substance use, and exiting homelessness. AB - This study tested whether social affiliation was associated with exits from homelessness for a county-wide probability sample of 397 homeless adults who were followed over a 15-month period culminating in 1992. For the total sample, support from family or friends and service use were related to an increased likelihood of exiting from homelessness. Surprisingly, exits from homelessness were associated with social affiliation (i.e., support from family/friends and services use) only among homeless adults who did not have current substance-use disorders. This relationship did not hold for those with current substance-use disorders. Findings suggest that homeless adults without current substance-use disorders may be better able to engage services and support from family/friends to exit homelessness than homeless adults who have current substance-use disorders. Perhaps service providers who are targeting homeless adults with substance-use disorders and want to help them exit homelessness need to emphasize initiating substance-use treatment before addressing other issues. PMID- 12747399 TI - Alcohol use and other psychiatric disorders in the formerly homeless and never homeless: prevalence, age of onset, comorbidity, temporal sequencing, and service utilization. AB - Interview survey data were collected on a large (n = 4730) general population sample of adults subsequently classified as "never homeless" (NH) or "formerly homeless" (FH), with the latter group consisting of persons who had past experience of at least a one-month period with no regular place to live. The objective was to analyze differences, as a function of this classification, in the prevalence, age of onset, comorbidity, temporal sequencing, and service utilization pertinent to alcohol-use and other psychiatric disorders. Almost half of the FH group were found to have a one-year DSM diagnosis, nearly twice the rate seen in the NH group. Moreover, at 15.1%, the prevalence of alcohol-use disorder (AUD) comorbid with one or more other psychiatric disorders was five times that reported by NH participants. Subsequent analyses addressed differences betweenthe FH (n = 167) and NH (n = 1031) groups within the subset who met criteria for one or more psychiatric diagnoses. Focusing on drinking behavior, we found that among the FH, dual-diagnosis was associated with elevated rates of alcohol-use problem symptoms and with greater alcohol consumption than were evident in the FH with AUD alone. Also, among the FH with comorbid AUD, as well as among those with two or more psychiatric disorders unrelated to alcohol, there was an earlier onset of psychiatric disorders than in the NH. This earlier onset may have placed these individuals at greater risk for later homelessness and AUD, and may also be indicative of a more severe course of illness. Differences between the FH and the NH suggest the importance of devoting special attention to this unique sample. PMID- 12747400 TI - Homelessness, substance misuse, and access to public entitlements in a soup kitchen population. AB - The study examined the effects of homelessness on access to public entitlements (Medicaid and food stamp programs) in a soup kitchen population. Data were collected between 1997 and 1999 from a sample of 343 adults at two soup kitchen sites in New York City. Five hypotheses, focusing on the effects of housing status (literal homelessness, unstable housing, and domiciled), frequency of drug/heavy alcohol use, drug/alcohol-user treatment history and childcare responsibilities on access to Medicaid and food stamp programs were tested. Multiple logistic regression analysis indicated that both literal homelessness and unstable housing were associated with less access to Medicaid and food stamps. Other significant findings were: current drug/alcohol-user treatment experience was associated with greater access to both Medicaid and food stamps, frequency of drug/heavy alcohol use was associated with less access to Medicaid only, and caring for children was associated with greater access to food stamps only. These findings support the crucial role of housing status in mediating access to entitlements, and the importance of drug/alcohol-user treatment involvement as a cue to seeking entitlements. The need to reduce health disparities through active and sustained outreach programs designed to enhance homeless persons' access to Medicaid and food stamp programs was discussed. PMID- 12747401 TI - Violence, homelessness, and HIV risk among crack-using African-American women. AB - This study compares the characteristics of out-of-treatment, homeless, crack using African-American women with those who are not homeless to determine what risks and protective factors differentiate the two groups. From 1999 to 2001, 683 out-of-treatment, African-American crack-using women (of whom 219 were categorized as homeless) were interviewed and serologically tested. Risk factors that were examined include adverse childhood experiences, psychological distress, physical health, violence and victimization, drug use, and risky sex behaviors. Protective factors that were examined include marital status, education, public assistance, and the responsibility of caring for children. Overall, both groups of women started crack use in their mid-twenties and started drug use with alcohol in their teenage years, though differed significantly on each risk factor examined. Logistic regression analysis found that variables associated with increased odds of being homeless are physical abuse before age 18, crack runs greater than 24 hours, income less than dollars 500 in the last 30 days, depression, and current cigarette smoking. Protective factors found are marital status, living with children under 18, having had a physical in the past year, and receiving money from welfare in the last 30 days. Being sexually assaulted in the past 90 days was marginally associated with homelessness in the model. These findings, specific to crack-using African-American women, suggest that not only do these women overall report painful histories and currently stressful lives, but homeless women are more likely than women who are not homeless to have experienced childhood abuse and are more involved with drug use. Interventions designed for these women need to consider gender, cultural, and contextual issues that not only incorporate aspects of risk reduction related to violence, alcohol use, and comorbid conditions, but also linkages that will address housing issues, education, and skills for independence. PMID- 12747402 TI - Substance use and delinquency during adolescence: a prospective look at an at risk sample. AB - This paper focuses on the relationship between adolescent substance use and delinquent behavior in a sample of homeless young people. Confirmatory factor analyses indicated that delinquency and substance use are best described as discrete factors, and competing theoretical models of the longitudinal association between these two factors were examined using structural equations modeling techniques. The results suggest that delinquent behavior is associated with changes in alcohol, marijuana, and drug use across time. This effect was statistically significant over relatively brief lags in time of six months or less. Combined with previous results, these findings challenge the utility of single-factor explanations of adolescent deviance for at-risk populations and suggest that the relationship between substance use and externalizing across time may be more dynamic than previously thought. Implications for intervention are also discussed. PMID- 12747403 TI - Sequential progression of substance use among homeless youth: an empirical investigation of the gateway theory. AB - We examined the sequence of substance-use initiation in 375 street youth (age 13 21) who were interviewed from 1994-99 in Seattle, Washington. Based on the "gateway theory," participants were categorized into six profiles to describe the order in which they initiated use of various substances (i.e., alcohol, marijuana, other drugs), or classified as nonprogressors if they had not tried all three classes of drugs. Youth progressing in the hypothesized gateway order (i.e., alcohol preceding marijuana, followed by other drugs) initiated their use at an earlier age than youth who had not progressed through all three substance classes. However, there was no relationship between a substance initiation profile and current substance-use. Implications include the recognition that street youth may follow different patterns of use than normative groups, and that interventions geared toward youth who use substances heavily must include contextual factors, in addition to substance-use history. PMID- 12747404 TI - Analysis of conserved non-rRNA genes of Tropheryma whipplei. AB - The causative agent of Whipple's disease, Tropheryma whipplei, is a slow-growing bacterium that remains poorly-understood. Genetic characterization of this organism has relied heavily upon rRNA sequence analysis. Pending completion of a complete genome sequencing effort, we have characterized several conserved non rRNA genes from T. whipplei directly from infected tissue using broad-range PCR and a genome-walking strategy. Our goals were to evaluate its phylogenetic relationships, and to find ways to expand the strain typing scheme, based on rDNA sequence comparisons. The genes coding for the ATP synthase beta subunit (atpD), elongation factor Tu (tuf), heat shock protein GroEL (groEL), beta subunit of DNA dependent RNA polymerase (rpoB), and RNase P RNA (rnpB) were analyzed, as well as the regions upstream and downstream of the rRNA operon. Phylogenetic analyses with all non-rRNA marker molecules consistently placed T. whipplei within the class, Actinobacteria. The arrangement of genes in the atpD and rpoB chromosomal regions was also consistent with other actinomycete genomes. Tandem sequence repeats were found upstream and downstream of the rRNA operon, and downstream of the groEL gene. These chromosomal sites and the 16S-23S rRNA intergenic spacer regions were examined in the specimens of 11 patients, and a unique combination of tandem repeat numbers and spacer polymorphisms was found in each patient. These data provide the basis for a more discriminatory typing method for T. whipplei. PMID- 12747405 TI - Virulence factors in food, clinical and reference Enterococci: A common trait in the genus? AB - The occurrence of several virulence traits (cytolysin, adhesins and hydrolytic enzymes) was investigated in a collection of 164 enterococci, including food and clinical isolates (from human and veterinary origin), as well as type and reference strains from 20 enterococcal species. Up to fifteen different cyl genotypes were found, as well as silent cyl genes. The occurrence of the cyl operon and haemolytic potential seems to be widespread in the genus. A significant association of this virulent trait with clinical isolates was found (p < 0.05). High levels of incidence were also observed for genes encoding surface adhesins (esp, efaA(fs), efaA(fm)), agg and gelE, irrespectively of species allocation and origin of strains. Although gelE behaves as silent in the majority of the strains, gelatinase activity predominates in clinical isolates, whereas lipase and DNase were mainly detected in food isolates pointing to their minor role as virulence determinants. No hyaluronidase activity was detected for all strains. Numerical hierarchic data analysis grouped the strains in three main clusters, two of them including a total of 50 strains with low number of virulence determinants (from 2 to 7) and the other with 114 strains with a high virulence potential (up to 12 determinants). No statistical association was found between virulence clusters and species allocation (p > 0.10), strongly suggesting that virulence determinants are a common trait in the genus Enterococcus. Clinical strains seem to be significantly associated with high virulence potential, whereas food, commensal and environmental strains harbour fewer virulence determinants (p < 0.01). A high level of relative diversity in virulence patterns was observed (Shannon's index varies from 0.95 to 1.0 among clusters), reinforcing the strain-specific nature of the association of virulence factors. Although a low risk seems to be associated with the use of enterococci in long-established artisanal cheeses, screening of virulence traits and their cross-synergies must be performed, particularly for commercial starters, probiotic strains and products to be used by high risk population groups. PMID- 12747406 TI - Teichococcus ludipueritiae gen. nov. sp. nov., and Muricoccus roseus gen. nov. sp. nov. representing two new genera of the alpha-1 subclass of the Proteobacteria. AB - Two bacterial isolates (170/96T and 173/96T) were recovered from the indoor building materials of a children's day care center. Phylogenetic analyses using the 16S rRNA gene sequences of both isolates indicated they both represent new lineages in the alpha-1-subclass of the Proteobacteria, with the highest sequence similarities of 93.7% and 93.6%, respectively to the type strain of Paracraurococcus ruber. When directly compared both isolates showed a 93.4% sequence similarity of their 16S rRNAs. The major respiratory quinone in both strains was a ubiquinone with 10 isoprenoid units and the major whole cell fatty acid of both strains was 18:1 omega7c. Both isolates also contained 18:1 2-OH and other fatty acids typical for members of the alpha-1 subclass of the Proteobacteria. Both strains were heterotrophic and strictly aerobic and formed slightly red-colored colonies on tryptone soy agar. Bacteriochlorophyll a could not be detected by direct spectrophotometric analyses of aerobically grown cells. On the basis of the phylogenetic analyses, physiological and biochemical characteristics, we propose that strains 170/96T and 173/96T represent two new genera and new species of the alpha-1 subclass of the Proteobacteria for which we propose the names Teichococcus ludipueritiae gen. nov. sp. nov., and Muricoccus roseus gen. nov. sp. nov., respectively. PMID- 12747407 TI - Staphylococcus equorum subsp. linens, subsp. nov., a starter culture component for surface ripened semi-hard cheeses. AB - Two staphylococcal strains, RP29T and RP33, were isolated from the main microflora of a surface ripened Swiss mountain cheese made from raw milk. These two strains were differentiated from the most closely related species Staphylococcus equorum on the basis of DNA-DNA hybridisation and phenotypic characteristics and are proposed as Staphylococcus equorum subsp. linens subsp. nov. They could be distinguished phenotypically from S. equorum by their sensitivity to all 14 tested antibiotics, especially to novobiocin, their incapability to ferment alpha-D-lactose, maltose, sucrose, D-trehalose, D-xylose, L-arabinose, salicin, D-ribose, D-raffinose, D-mannitol, and D-alanine. The GenBank accession numbers for the reference sequences of the 16S rDNA and the hsp60 gene used in this study are AF527483 and AF527484, respectively. 30 tons of a semi-hard Swiss cheese were produced with Staphylococcus equorum subsp. linens DSM 15097T as starter culture component in addition to Debaryomyces hansenii, Geotrichum candidum, Brevibacterium linens, Corynebacterium casei for surface ripened cheeses. The products were sensorically and hygienically perfect. Therefore, Staphylococcus equorum subsp. linens DSM 15097T can be proposed as starter culture component for surface ripened cheeses without any detected antibiotic resistances. The type strain of Staphylococcus equorum subsp. linens is DSM 15097T (CIP 107656T). PMID- 12747408 TI - Three new antibiotic producing species of the genus Amycolatopsis, Amycolatopsis balhimycina sp. nov., A. tolypomycina sp. nov., A. vancoresmycina sp. nov., and description of Amycolatopsis keratiniphila subsp. keratiniphila subsp. nov. and A. keratiniphila subsp. nogabecina subsp. nov. AB - Screening of Indian soils for strains producing antibiotics active against methicillin resistant staphylococci resulted in the isolation of the actinomycetes FH 1894T, ST 101170T, and FH 1893T, producing novel glycopeptide antibiotics and a polyenic compound, respectively. Studies of the cellular fatty acids and the absence of mycolic acids as well as the 16S rRNA gene sequence analysis of the producers indicated their membership to the genus Amycolatopsis. Comparison of genomic and metabolic properties of these strains with known species of this genus indicated that they represent new species for which the names Amycolatopsis balhimycina (type strain FH 1894T, DSM 44591T) and Amycolatopsis vancoresmycina (type strain ST 101170T, DSM 44592T) are proposed. The producer of the antibiotic Tolypomycin, strain IFO 14664T, representing the invalid species "Amycolatopsis tolypophorus", was also included in this investigation. As this strain was found to represent a valid species of the genus Amycolatopsis the name Amycolatopsis tolypomycina is proposed for this species, with the type strain IFO 14664T (DSM 44544T). The producer of the antibiotic Nogabecin, strain FH 1893T, was found to be closely related to the type strain of A. keratiniphila DSM 44409T which justifies the description of two subspecies of this species, A. keratiniphila subsp. keratiniphila DSM 44409T and A. keratiniphila subsp. nogabecina DSM 44586T. PMID- 12747409 TI - Description of Devosia neptuniae sp. nov. that nodulates and fixes nitrogen in symbiosis with Neptunia natans, an aquatic legume from India. AB - Neptunia natans is a unique aquatic legume indigenous to tropical and sub tropical regions and is nodulated symbiotically by rhizobia using an unusual infection process unlike any previously described. Previously, isolates of neptunia-nodulating rhizobia from Senegal were characterized as Allorhizobium undicola. Here we report on a different group of neptunia-nodulating rhizobia isolated from India. Sequencing of the 16S rDNA gene from two of these Indian isolates (strains J1T and J2) show that they belong in the genus Devosia rather than Allorhizobium. Currently, the only described Devosia species is D. riboflavina (family Hyphomicrobiaceae, order Rhizobiales). The complete 16S rDNA sequences of strains J1T and J2 are 95.9% homologous to the type strain, D. riboflavina LMG 2277T, suggesting that these neptunia-nodulating strains from India belong to a new Devosia species. This hypothesis was confirmed by further studies of polyphasic taxonomy (DNA-DNA hybridisation, TP-RAPD patterns, SDS-PAGE of cellular proteins, 16S rDNA RFLP patterns, carbon source utilisation, cellular fatty acid analysis and other phenotypic characterisations), all of which support the proposal that these neptunia-nodulating strains constitute a new Devosia species, which we name Devosia neptuniae sp. nov. These gram negative, strictly aerobic short rods are motile by a subpolar flagellum, positive for catalase, oxidase, urease and beta-galactosidase, can utilise several carbohydrates (but not organic acids) as carbon sources and contain C18:0 3-OH, cis-7 C18:1 11 methyl and cis-7 C18:1 as their major cellular fatty acids. Unlike D. riboflavina, the longer-chain C24:1 3-OH and C26:1 3-OH hydroxy fatty acids are not detected. The type strain of D. neptuniae is LMG 21357T (CECT 5650T). Assignment of this new taxon represents the fourth example in the literature of a non-rhizobial genus of bacteria capable of forming a bonafide dinitrogen-fixing root-nodule symbiosis with legume plants. PMID- 12747410 TI - Sinorhizobium americanus sp. nov., a new Sinorhizobium species nodulating native Acacia spp. in Mexico. AB - The sinorhizobia isolated from root nodules of Acacia species native of Mexico constitute a diverse group of bacteria on the basis of their metabolic enzyme electromorphs but share restriction patterns of the PCR products of 16S rRNA genes and a common 500 kb symbiotic plasmid. They are distinguished from other Sinorhizobium species by their levels of DNA-DNA hybridization and the sequence of 16S rRNA and nifH genes. nolR gene hybridization patterns were found useful to identify sinorhizobia and characterize species. A new species, Sinorhizobium americanus, is described and the type strain is CFNEI 156 from Acacia acatlensis. PMID- 12747411 TI - Vibrio tasmaniensis sp. nov., isolated from Atlantic salmon (Salmo salar L.). AB - We describe the polyphasic characterization of four Vibrio isolates which formed a tight AFLP group in a former study. The group was closely related to V. cyclitrophicus, V. lentus and V. splendidus (98.2-98.9% similarity) on the basis of the 16S rDNA sequence analysis, but by DNA-DNA hybridisation experiments it had at maximum 61% DNA similarity towards V. splendidus. Thus, we propose that the isolates represent a new Vibrio species i.e. V. tasmaniensis (LMG 20012T; EMBL under the accession numbers AJ316192; mol% G+C of DNA of the type strain is 44.7). Useful phenotypical features for discrimination of V. tasmaniensis from other Vibrio species include gelatinase and beta-galactosidase activity, fatty acid composition (particularly 14:0), utilisation and fermentation of different compounds (e.g. sucrose, melibiose and D-galactose) as sole carbon source. PMID- 12747412 TI - Thermomonas hydrothermalis sp. nov., a new slightly thermophilic gamma proteobacterium isolated from a hot spring in central Portugal. AB - Several non-pigmented bacterial isolates, with an optimum growth temperature of about 50 degrees C, were recovered from the hot spring at Sao Gemil in Central Portugal. Phylogenetic analyses of the 16S rRNA gene sequence of strain SGM-6T indicated that this organism represents a new species of the gamma-subclass of the Proteobacteria that is closely related to the newly described slightly thermophilic species Thermomonas haemolytica. The major fatty acids of strains SGM-6T and SGM-7 are C15:0 iso, C16:0 iso, C11:0 iso and C11:0 iso 3OH. Ubiquinone 8 is the major respiratory quinone. The new isolates are strictly organotrophic and aerobic. Strain SGM-6T only assimilated D-glucose, D-maltose, D cellobiose, D-furanose, L-glutamate, L-glutamine, L-lysine, L-proline, L ornithine, acetate, L-glutamic acid and pyruvate of sixty-five carbon sources tested. Strain SGM-7 also assimilates L-serine, but does not assimilate L ornithine. On the basis of the phylogenetic analyses, physiological and biochemical characteristics, we propose that strains SGM-6T and SGM-7 represent a new species most closely related to Thermomonas haemolytica for which we propose the name Thermomonas hydrothermalis. PMID- 12747413 TI - Croceibacter atlanticus gen. nov., sp. nov., a novel marine bacterium in the family Flavobacteriaceae. AB - A bright, saffron-colored marine bacterium HTCC2559T was isolated from the Bermuda Atlantic Time Series station in the western Sargasso Sea, Atlantic Ocean by high throughput culturing methods and characterized by polyphasic approaches. Phenotypic data and phylogenetic analyses showed that the strain is a member of the family Flavobacteriaceae. The strain was gram-negative, non-motile, chemoheterotrophic, strictly aerobic, NaCl-requiring, rod-shaped cells that contain carotenoid pigments but not flexirubin. Several kinds of macromolecules (gelatin, DNA, starch, casein, and elastin) were degraded and carbohydrates, sugar alcohols, organic acids, and amino acids were utilized as sole carbon sources. The dominant fatty acids were branched or hydroxy acids, and 3-OH i17:0, i15:0, i15:1, and i17:1 omega9c were abundant. The DNA G+C content of the strain is 34.8 mol%. Phylogenetic analyses using three treeing algorithms based on 16S rRNA gene sequences revealed that the strain formed a very distinct lineage that is allied closely with several seawater environmental clones in the family Flavobacteriaceae. Therefore, it is proposed from the polyphasic studies that strain HTCC2559T (=ATCC BAA-628T = KCTC 12090T) belongs to a new genus and species named Croceibacter atlanticus gen. nov., sp. nov. PMID- 12747414 TI - Clostridium bolteae sp. nov., isolated from human sources. AB - Seven obligately anaerobic, gram-positive, rod-shaped, spore-forming organisms isolated from human sources were characterized using phenotypic and molecular taxonomic methods. Comparative 16S rRNA gene sequencing showed that the strains were genetically highly related to each other (displaying >99% sequence similarity) and represent a previously unknown sub-line within the Clostridium coccoides rRNA group of organisms. Strains of the unidentified bacterium used carbohydrate as fermentable substrates, producing acetic acid and lactic acid as the major products of glucose metabolism. The closest described species to the novel bacterium corresponded to Clostridium clostridioforme, although a 16S rRNA sequence divergence of 3% demonstrated they represent different species. Genomic DNA-DNA pairing studies confirmed the separateness of the unknown species and Clostridium clostridioforme. Based on phenotypic and phylogenetic evidence, it is therefore proposed that the unknown bacterium, be classified as Clostridium bolteae sp. nov. The type strain of Clostridium bolteae is WAL 16351T (= ATCC(T) = BAA-613T, CCUG(T) = 46953T). PMID- 12747415 TI - Genotypic differentiation of twelve Clostridium species by polymorphism analysis of the triosephosphate isomerase (tpi) gene. AB - Housekeeping genes encoding metabolic enzymes may provide alternative markers to 16S ribosomal DNA (rDNA) for genotypic and phylogenetic characterization of bacterial species. We have developed a PCR-restriction fragment length polymorphism (PCR-RFLP) assay, targeting the triosephosphate isomerase (tpi) gene, which allows the differentiation of twelve pathogenic Clostridium species. Degenerate primers constructed from alignments of tpi sequences of various gram positive bacteria allowed the amplification of a 501 bp target region in the twelve Clostridium type strains. A phylogenetic tree constructed from the nucleotidic sequences of these tpi amplicons was well correlated with that inferred from analysis of 16S rDNA gene sequences. The analysis of tpi sequences revealed restriction sites of enzyme AluI that could be species-specific. Indeed, AluI digestion of amplicons from the twelve type strains provided distinct restriction patterns. A total of 127 strains (three to sixteen strains for each species) was further analyzed by PCR-RFLP of the tpi gene, and confirmed that each species could be characterized by one to three restriction types (RTs). The differences between RTs within species could be explained by point mutations in AluI restriction sites of the tpi sequences. PCR-restriction analysis of the tpi gene offers an accurate tool for species identification within the genus Clostridium, and provides an alternative marker to 16S rDNA for phylogenetic analyses. PMID- 12747416 TI - Inter- and intraspecies variations of the 16S-23S rDNA intergenic spacer region of various streptococcal species. AB - The 16S-23S rDNA intergenic spacer regions (ISR) of different streptococcal species and subspecies were amplified with primers derived from the highly conserved flanking regions of the 16S rRNA and 23S rRNA genes. The single sized amplicons showed a uniform pattern for S. agalactiae, S. dysgalactiae subsp. dysgalactiae (serogroup C), S. dysgalactiae subsp. equisimilis (serogroup G), S. dysgalactiae subsp. dysgalactiae (serogroup L), S. canis, S. phocae, S. uberis, S. parauberis, S. pyogenes and S. equi subsp. equi, respectively. The amplicons of S. equi subsp. zooepidemicus, S. porcinus and S. suis appeared with 3, 5 and 3 different sizes, respectively. ISR of selected strains of each species or subspecies investigated were sequenced and multiple aligned. This allowed a separation of ISR into regions, with 7 regions for S. agalactiae, S. dysgalactiae subsp. dysgalactiae (serogroup C), S. dysgalactiae subsp. equisimilis (serogroup G), S. dysgalactiae subsp. dysgalactiae (serogroup L), S. canis, S. phocae, S. pyogenes and S. suis, 8 regions for S. uberis and S. parauberis and mostly 9 regions for S. equi subsp. equi, S. equi subsp. zooepidemicus and S. porcinus. Region 4, encoding the transfer RNA for alanine (tRNA(Ala)), was present and identical for all isolates investigated. The size and sequence of ISR appears to be a unique marker for streptococci of various species and subspecies and could be used for bacterial identification. In addition the size and sequence variations of ISR of S. equi subsp. zooepidemicus, S. porcinus and S. suis allows a molecular typing of isolates of these species possibly useful in epidemiological aspects. PMID- 12747417 TI - Morphologies and phylogenetic classification of cellulolytic myxobacteria. AB - The evolutionary distances of the 16S rDNA sequences in cellulolytic myxobacteria are less than 3%, which units all the strains into a single genus, Sorangium. The size of myxospores and the shape of sporangioles, rather than fruiting body colors or swarm morphologies are consistent with the changes of the 16S rDNA sequences. It is suggested that there are at least two species in the genus Sorangium: one includes strains with small myxospores and spherical sporangioles, and the color of the fruiting bodies is normally orange or brown, though sometimes yellow or black. The second species has large myxospores, polyhedral sporangioles with many inter-cystic substrates, and normally deep brown to black color. PMID- 12747418 TI - Enumeration of Bacteroides species in human faeces by fluorescent in situ hybridisation combined with flow cytometry using 16S rRNA probes. AB - Bacteroides is a predominant group of the faecal microbiota in healthy adults. To investigate the species composition of Bacteroides by fluorescent in situ hybridisation (FISH) combined with flow cytometry, we developed five species specific probes targeting the 16S rRNA. Probes were designed to identify cells belonging to Bacteroides distasonis, B. fragilis, B. ovatus, B. vulgatus and B. putredinis. The species-specificity of the probes was assessed against a collection of reference strains from the Cytophaga-Flavobacterium-Bacteroides group. The results of the FISH experiments showed that the probes were specific as they only detected strains of the target species. Determining the fluorescence intensity of each probe relative to that of the EUB 338 probe (domain bacteria) further showed that each species probe easily accessed the targeted site. The probes were applied to quantify the Bacteroides species in faeces collected from 20 healthy adults. All five species were detected in the faecal samples. Cells hybridised with Bfra 998 were the most frequent as they were observed in 90% of individuals (18/20 samples, mean proportion of 3.9 +/- 2.2%). The cells hybridised with Bvulg 1017 were observed in 85% of individuals (17/20 samples) and represented with a mean proportion of 4.2 +/- 6.1%, the most abundant Bacteroides species in human faeces. Cells hybridising with probes for B. ovatus, B. distasonis and B. putredinis were less frequently detected. The large distribution of B. vulgatus and B. fragilis in human faeces is in accordance with previous reports based on culture or molecular studies. This work showed that fluorescent in situ hybridisation is a tool appropriate for a high-resolution analysis of the species composition of complex ecosystems and especially of the Bacteroides group within the faecal microbiota. PMID- 12747419 TI - Molecular characterisation and antimicrobial resistance of Vibrio vulnificus and Vibrio alginolyticus isolated from mussels (Mytilus galloprovincialis). AB - Twelve Vibrio vulnificus biotype 1 and 11 Vibrio alginolyticus isolated from mussels in Italy were analysed by antimicrobial resistance, plasmid profiles, random amplification of polymorphic DNA (RAPD), and single enzyme amplified fragment length polymorphism (sAFLP). Plasmid DNA was detected in three V. vulnificus and four V. alginolyticus cultures. All isolates were resistant to at least two antimicrobial agents: all isolates were resistant to ampicillin, carbenicillin and streptomycin, except one V. alginolyticus which was sensitive to carbenicillin and two V. alginolyticus which were sensitive to streptomycin. No association was detected between the presence of plasmid DNA and antimicrobial resistance. Seven of the twelve V. vulnificus and two of the eleven V. alginolyticus cultures were susceptible to the 10 microg of the vibriostatic compound O/129; all cultures were susceptible to the 150 microg of O/129. Both RAPD and sAFLP was found to be reproducible. Ten sAFLP and seven RAPD profiles were detected amongst the 12 V. vulnificus cultures: three cultures were identified as indistinguishable by both methods. RAPD and sAFLP analysis of V. alginolyticus generated nine and seven profiles respectively, and these two methods were independent. These results demonstrate extreme variability of V. vulnificus and V. alginolyticus isolated from mussels, and both RAPD and sAFLP provided information on intraspecific differences which will be useful for molecular epidemiological or ecological studies. A combination of methods gave optimal discrimination, although a single method could provide sufficient information to characterise V. vulnificus isolates. PMID- 12747420 TI - Recovery of a new biogroup of Yersinia ruckeri from diseased rainbow trout (Oncorhynchus mykiss, Walbaum). AB - Cultures of a new biogroup of Yersinia ruckeri, the causal agent of enteric redmouth (ERM), were recovered in England from diseased rainbow trout (Oncorhynchus mykiss, Walbaum), which had been previously vaccinated with a commercial ERM vaccine. The bacterial isolates were confirmed as Y. ruckeri by the results of sequencing the 16S rRNA, but differed from the characteristics of the taxon by positivity for the Voges Proskauer reaction and a general lack of motility, and could not be equated with any of the existing serovars. Cultures were pathogenic in laboratory-based infectivity experiments with 100% mortalities occurring in juvenile rainbow trout (average weight = 10 g) within 4-days of intraperitoneal or intramuscular injection with 10(5) cells/fish. Protection against disease was achieved using a formalin-inactivated whole vaccine prepared against a representative isolate. PMID- 12747421 TI - Different Yersinia enterocolitica 4:O3 genotypes found in pig tonsils in Southern Germany and Finland. AB - The distribution of different genotypes of Y. enterocolitica 4:O3 strains recovered from pig tonsils in Southern Germany and Finland in 1999-2000 was investigated. A total of 96 and 207 Y. enterococolitica 4:O3 isolates recovered from 47 and 66 tonsils of finishing pigs in Germany and Finland, respectively, were characterised with PFGE using NotI enzyme. In all, 39 different NotI profiles were obtained, only one of which, NB1, was found in both Germany and Finland. All strains were further characterised with ApaI and XhoI enzymes. When the 54 German and 74 Finnish strains were characterised with all three enzymes, 51 genotypes were obtained. The 23 genotypes found in German strains differed from the 28 found in Finnish strains. These results indicate that Y. enterocolitica 4:O3 genotypes have a differential geographical distribution and thus can be used in epidemiological studies. PMID- 12747422 TI - Molecular typing of Aspergillus ochraceus and construction of species specific SCAR-primers based on AFLP. AB - 70 strains of Aspergillus ochraceus mainly isolated from Brazilian coffee related sources were investigated for genetic relatedness using automated laser fluorescence analysis of AFLP fragments. Cluster analysis of fingerprints revealed a very close relationship among most of the strains. Based on these results, a sub-set of characteristic A. ochraceus strains was chosen for the detection of marker sequences. These sequences were obtained from silver stained AFLPs separated on polyacrylamide gels. A number of bands characteristic for A. ochraceus were detected and cut out from the gels. DNA was reamplified, cloned and fragments were sequenced. Based on these sequences a set of SCAR PCR-primers was constructed. PCRs were optimised for specificity and subsequently tested against a panel of Aspergillus species. Using this approach a PCR specific for Aspergillus ochraceus was developed. PMID- 12747423 TI - Validation of an official control method for enumeration of authorised probiotic yeast in animal feed. AB - An official control method in the framework of Council Directive 70/524/EEC for probiotic yeast used as feed additives was validated in a collaborative study by twenty laboratories in 12 European Countries. A pour plate method following ISO 7954 using chloramphenicol glucose yeast extract (CGYE) and a plate count method using CHROMagar Candida were used. Precision data in terms of repeatability (r) and reproducibility (R) of the method using different feeding stuffs and three inoculation levels were determined. Yeast was present in the samples in mixtures with other probiotic feed additives at a lower, a higher concentration or not present. The enumeration of yeast on CGYE agar showed for the lower and higher concentration a RSD(r) of 2.4-4.9% and a RSD(R) of 7.7-8%, respectively and was preferred by the majority of labs. CHROMagar Candida had a RSD(r) of 1.9-2.8% and a RSD(R) of 1.9-5.9%. For routine analysis the use of the pour plate technique is recommended. CHROMagar Candida can be used for confirmation of the species Saccharomyces cerevisiae. The methods are not recommended for mineral feeds. The results from this study are intended for consideration for adoption as CEN and ISO standards. PMID- 12747424 TI - Ultrasonic imaging of myocardial strain using cardiac elastography. AB - Clinical assessment of myocardial ischemia based on visually-assessed wall motion scoring from echocardiography is semiquantitative, operator dependent, and heavily weighted by operator experience and expertise. Cardiac motion estimation methods such as tissue Doppler imaging, used to assess myocardial muscle velocity, provides quantitative parameters such as the strain-rate and strain derived from Doppler velocity. However, tissue Doppler imaging does not differentiate between active contraction and simple rotation or translation of the heart wall, nor does it differentiate tethering (passively following) tissue from active contraction. In this paper, we present a strain imaging modality called cardiac elastography that provides two-dimensional strain information. A method for obtaining and displaying both directional and magnitude cardiac elastograms and displaying strain over the entire cross-section of the heart is described. Elastograms from a patient with coronary artery disease are compared with those from a healthy volunteer. Though observational, the differences suggest that cardiac elastography may be a useful tool for assessment of myocardial function. The method is two-dimensional, real time and avoids the disadvantage of observer-dependent judgment of myocardial contraction and relaxation estimated from conventional echocardiography. PMID- 12747425 TI - Tissue-mimicking oil-in-gelatin dispersions for use in heterogeneous elastography phantoms. AB - A ten-month study is presented of materials for use in heterogeneous elastography phantoms. The materials consist of gelatin with or without a suspension of microscopic safflower oil droplets. The highest volume percent of oil in the materials is 50%. Thimerosal acts as a preservative. The greater the safflower oil concentration, the lower the Young's modulus. Elastographic data for heterogeneous phantoms, in which the only variable is safflower oil concentration, demonstrate stability of inclusion geometry and elastic strain contrast. Young's modulus ratios (elastic contrasts) producible in a heterogeneous phantom are as high as 2.7. The phantoms are particularly useful for ultrasound elastography. They can also be employed in MR elastography, although the highest achievable ratio of longitudinal to transverse relaxation times is considerably less than is the case for soft tissues. PMID- 12747426 TI - Colorimetry for CRT displays. AB - We analyze the sources of error in specifying color in CRT displays. These include errors inherent in the use of the color matching functions of the CIE 1931 standard observer when only colorimetric, not radiometric, calibrations are available. We provide transformation coefficients that prove to correct the deficiencies of this observer very well. We consider four different candidate sets of cone sensitivities. Some of these differ substantially; variation among candidate cone sensitivities exceeds the variation among phosphors. Finally, the effects of the recognized forms of observer variation on the visual responses (cone excitations or cone contrasts) generated by CRT stimuli are investigated and quantitatively specified. Cone pigment polymorphism gives rise to variation of a few per cent in relative excitation by the different phosphors--a variation larger than the errors ensuing from the adoption of the CIE standard observer, though smaller than the differences between some candidate cone sensitivities. Macular pigmentation has a larger influence, affecting mainly responses to the blue phosphor. The estimated combined effect of all sources of observer variation is comparable in magnitude with the largest differences between competing cone sensitivity estimates but is not enough to disrupt very seriously the relation between the L and M cone weights and the isoluminance settings of individual observers. It is also comparable with typical instrumental colorimetric errors, but we discuss these only briefly. PMID- 12747427 TI - Analytical solution for irradiance due to inhomogeneous Lambertian polygonal emitters. AB - We present an analytic solution for the irradiance at a point due to a polygonal Lambertian emitter with radiant exitance that varies with position according to a polynomial of arbitrary degree. This is a basic problem that arises naturally in radiative transfer and more specifically in global illumination, a subfield of computer graphics. Our solution is closed form except for a single nonalgebraic special function known as the Clausen integral. We begin by deriving several useful formulas for high-order tensor analogs of irradiance, which are natural generalizations of the radiation pressure tensor. We apply the resulting tensor formulas to linearly varying emitters, obtaining a solution that exhibits the general structure of higher-degree cases, including the dependence on the Clausen integral. We then generalize to higher-degree polynomials with a recurrence formula that combines solutions for lower-degree polynomials; the result is a generalization of Lambert's formula for homogeneous diffuse emitters, a well known formula with many applications in radiative transfer and computer graphics. Similar techniques have been used previously to derive closed-form solutions for the irradiance due to homogeneous polygonal emitters with directionally varying radiance. The present work extends this previous result to include inhomogeneous emitters, which proves to be significantly more challenging to solve in closed form. We verify our theoretical results with numerical approximations and briefly discuss their potential applications. PMID- 12747428 TI - Cramer-Rao analysis of orientation estimation: viewing geometry influences on the information conveyed by target features. AB - A methodology for analyzing an imaging sensor's ability to assess target properties is developed. By the application of a Cramer-Rao covariance analysis to a statistical model relating the sensor measurements to the target, a lower bound can be calculated on the accuracy with which any unbiased algorithm can form estimates of target properties. Such calculations are important in understanding how a sensor's design influences its performance for a given assessment task and in performing feasibility studies or system architecture design studies between sensor designs and sensing modalities. A novel numerical model relating a sensor's measurements to a target's three-dimensional geometry is developed in order to overcome difficulties in accurately performing the required numerical computations. The accuracy of the computations is verified against simple test cases that can be solved in closed form. Examples are presented in which the approach is used to investigate the influence of viewing perspective on orientation accuracy limits. These examples are also used to examine the potential accuracy improvement that could be gained by fusing multiperspective data. PMID- 12747429 TI - Cramer-Rao analysis of orientation estimation: influence of target model uncertainties. AB - We explore the use of Cramer-Rao bound calculations for predicting fundamental limits on the accuracy with which target characteristics can be determined by using imaging sensors. In particular, estimation of satellite orientation from high-resolution sensors is examined. The analysis role that such bounds provide for sensor/experiment design, operation, and upgrade is discussed. Emphasis is placed on the importance of including all relevant target/sensor uncertainties in the analysis. Computer simulations are performed that illustrate that uncertainties in target features (e.g., shape, reflectance, and relative orientation) have a significant impact on the bounds and provide considerable insight as to how details of the three-dimensional target structure may influence the estimation process. The simulations also address the impact that a priori information has on the bounds. PMID- 12747430 TI - Rigorous theory of the diffraction of Gaussian beams by finite gratings: TE polarization. AB - A rigorous modal theory for the diffraction of Gaussian beams from N equally spaced slits (finite grating) in a planar perfectly conducting thin screen is presented. The case of normal incidence and TE polarization state is considered; i.e., the electric field is parallel to the slits. The characteristics of the far field diffraction patterns, the transmission coefficient, and the normally diffracted energy as a function of several optogeometrical parameters are analyzed within the so-called vectorial region, where the polarization effects are important. The diffraction pattern of an aperiodic grating is also considered. In addition, one diffraction property known to be valid in the scalar region is generalized to the vectorial region: the existence of constant intensity angles in the far field when the incident beam wave is scanned along the N slits. The classical grating equation is tested for incident Gaussian beams under several conditions. PMID- 12747432 TI - Parallel implementation of the biorthogonal multiresolution time-domain method. AB - The three-dimensional biorthogonal multiresolution time-domain (Bi-MRTD) method is presented for both free-space and half-space scattering problems. The perfectly matched layer (PML) is used as an absorbing boundary condition. It has been shown that improved numerical-dispersion properties can be obtained with the use of smooth, compactly supported wavelet functions as the basis, whereas we employ the Cohen-Daubechies-Fouveau (CDF) biorthogonal wavelets. When a CDF wavelet expansion is used, the spatial-sampling rate can be reduced considerably compared with that of the conventional finite-difference time-domain (FDTD) method, implying that larger targets can be simulated without sacrificing accuracy. We implement the Bi-MRTD on a cluster of allocated-memory machines, using the message-passing interface (MPI), such that very large targets can be modeled. Numerical results are compared with analytical ones and with those obtained by use of the traditional FDTD method. PMID- 12747431 TI - Surface plasmon polaritons on narrow-ridged short-pitch metal gratings in the conical mount. AB - Recent investigations into high-aspect-ratio short-pitch metal grating structures have shown that it is possible to excite surface plasmon polaritons (SPPs) even in the zero-order region of the spectrum. The predominant reason this is possible is that extremely large bandgaps occur in the SPP dispersion curves, which are caused by the large depths, and heights, of the structures. The form of the resultant dispersion curves has also been found to be highly dependent on the shape of the grating profile. We present an extension to a previously published paper that described the nature of the SPPs excited on narrow-ridged short-pitch metal gratings in the classical mount by considering the case in which the radiation is incident at nonzero azimuthal angles (the conical mount). In particular, we consider the case of 90 degrees and 45 degrees azimuthal angles and discuss the coupling to the SPP modes and the way in which polarization conversion is evident on such structures. PMID- 12747433 TI - Atmospheric optical communication with a Gaussian Schell beam. AB - We consider a wireless optical communication link in which the laser source is a Gaussian Schell beam. The effects of atmospheric turbulence strength and degree of source spatial coherence on aperture averaging and average bit error rate are examined. To accomplish this, we have derived analytic expressions for the spatial covariance of irradiance fluctuations and log-intensity variance for a Gaussian beam of any degree of coherence in the weak fluctuation regime. When spatial coherence of the transmitted source beam is reduced, intensity fluctuations (scintillations) decrease, leading to a significant reduction in the bit error rate of the optical communication link. We have also identified an enhanced aperture-averaging effect that occurs in tightly focused coherent Gaussian beams and in collimated and slightly divergent partially coherent beams. The expressions derived provide a useful design tool for selecting the optimal transmitter beam size, receiver aperture size, beam spatial coherence, transmitter focusing, etc., for the anticipated atmospheric channel conditions. PMID- 12747434 TI - Analytical models of optical refraction in the troposphere. AB - An extremely accurate but simple asymptotic description (with known error) is obtained for the path of a ray propagating over a curved Earth with radial variations in refractive index. The result is sufficiently simple that analytic solutions for the path can be obtained for linear and quadratic index profiles. As well as rendering the inverse problem trivial for these profiles, this formulation shows that images are uniformly magnified in the vertical direction when viewed through a quadratic refractive-index profile. Nonuniform vertical distortions occur for higher-order refractive-index profiles. PMID- 12747435 TI - State-estimation approach to the nonstationary optical tomography problem. AB - We propose a new numerical approach to the nonstationary optical (diffusion) tomography (OT) problem. The assumption in the method is that the absorption and/or diffusion coefficients are nonstationary in the sense that they may exhibit significant changes during the time that is needed to measure data for one traditional image frame. In the proposed method, the OT problem is formulated as a state-estimation problem. Within the state-estimation formulation, the absorption and/or diffusion coefficients are considered a stochastic process. The objective is to estimate a sequence of states for the process when the state evolution model for the process, the observation model for OT experiments, and data on the exterior boundary are given. In the proposed method, the state estimates are computed by using Kalman filtering techniques. The performance of the proposed method is evaluated on the basis of synthetic data. The simulations also illustrate that further improvements to the results in nonstationary applications can be obtained by adjustment of the measurement protocol. PMID- 12747436 TI - Inverse problem in optical diffusion tomography. III. Inversion formulas and singular-value decomposition. AB - We continue our study of the inverse scattering problem for diffuse light. In particular, we derive inversion formulas for this problem that are based on the functional singular-value decomposition of the linearized forward-scattering operator in the slab, cylindrical, and spherical geometries. Computer simulations are used to illustrate our results in model systems. PMID- 12747437 TI - Inverse problem in optical diffusion tomography. IV. Nonlinear inversion formulas. AB - We continue our study of the inverse scattering problem for diffuse light. In contrast to our earlier work, in which we considered the linear inverse problem, we now consider the nonlinear problem. We obtain a solution to this problem in the form of a functional series expansion. The first term in this expansion is the pseudoinverse of the linearized forward-scattering operator and leads to the linear inversion formulas that we have reported previously. The higher-order terms represent nonlinear corrections to this result. We illustrate our results with computer simulations in model systems. PMID- 12747438 TI - Theories for the design of a hybrid refractive-diffractive superresolution lens with high numerical aperture. AB - By geometrical optics and the Rayleigh-Sommerfeld diffraction formula, theories for the design of a hybrid refractive-diffractive superresolution lens (HRDSL) with high numerical aperture are constructed. Differences between the profile of the diffractive superresolution element (DSE) with high numerical aperture and that with low numerical aperture are indicated. Optimization theory can obtain a globally optimal solution through a linear programming much more simplified than the corresponding one in Liu et al. [J. Opt. Soc. Am. A 19, 2185 (2002)]. The rules of the structure of the designed DSE are both theoretically proved and numerically verified. Comparison of this optimization theory with the other design theories and examples of designing the HRDSL with high numerical aperture are provided. Last, some limits of optical superresolution with high numerical aperture are set and compared with those for low numerical aperture. PMID- 12747439 TI - General n-dimensional quadrature transform and its application to interferogram demodulation. AB - Quadrature operators are useful for obtaining the modulating phase phi in interferometry and temporal signals in electrical communications. In carrier frequency interferometry and electrical communications, one uses the Hilbert transform to obtain the quadrature of the signal. In these cases the Hilbert transform gives the desired quadrature because the modulating phase is monotonically increasing. We propose an n-dimensional quadrature operator that transforms cos(phi) into -sin(phi) regardless of the frequency spectrum of the signal. With the quadrature of the phase-modulated signal, one can easily calculate the value of phi over all the domain of interest. Our quadrature operator is composed of two n-dimensional vector fields: One is related to the gradient of the image normalized with respect to local frequency magnitude, and the other is related to the sign of the local frequency of the signal. The inner product of these two vector fields gives us the desired quadrature signal. This quadrature operator is derived in the image space by use of differential vector calculus and in the frequency domain by use of a n-dimensional generalization of the Hilbert transform. A robust numerical algorithm is given to find the modulating phase of two-dimensional single-image closed-fringe interferograms by use of the ideas put forward. PMID- 12747440 TI - Efficient excitation of self-collimated beams and single Bloch modes in planar photonic crystals. AB - Using finite-difference time-domain calculations, we investigate out-of-plane coupling between a square-lattice planar photonic crystal and a conventional waveguide located above the photonic crystal. We couple a waveguide oriented in the gamma(chi) direction to a photonic crystal mode in the second band and show that anticrossing takes place. In this way, a self-collimated beam is launched in the planar photonic crystal, with full power transfer. Furthermore, we investigate the coupling between a waveguide oriented in the gamma(mu) direction and a photonic crystal and show that single photonic crystal modes can be selectively excited. PMID- 12747441 TI - Three-dimensional optical-transfer-function analysis of fiber-optical two-photon fluorescence microscopy. AB - The three-dimensional optical transfer function is derived for analyzing the imaging performance in fiber-optical two-photon fluorescence microscopy. Two types of fiber-optical geometry are considered: The first involves a single-mode fiber for delivering a laser beam for illumination, and the second is based on the use of a single-mode fiber coupler for both illumination delivery and signal collection. It is found that in the former case the transverse and axial cutoff spatial frequencies of the three-dimensional optical transfer function are the same as those in conventional two-photon fluorescence microscopy without the use of a pinhole.However, the transverse and axial cutoff spatial frequencies in the latter case are 1.7 times as large as those in the former case. Accordingly, this feature leads to an enhanced optical sectioning effect when a fiber coupler is used, which is consistent with our recent experimental observation. PMID- 12747443 TI - Optimal beam splitters for the division-of-amplitude photopolarimeter. AB - Optimal optical parameters of the beam splitter that is used in the division-of amplitude photopolarimeter are determined. These are (1) 50%-50% split ratio of the all-dielectric beam splitter, (2) differential phase shifts in reflection and transmission delta(r) and delta(t) that differ by +/- pi/2, and (3) ellipsometric parameters (psi(r), psi(t)) = (27.368 degrees, 62.632 degrees) or (62.632 degrees, 27.368 degrees). It is also shown that for any nonabsorbing beam splitter that splits incident unpolarized light equally, the relationship psi(r) + psi(t) = pi/2 is always satisfied. PMID- 12747442 TI - Triply periodic bicontinuous structures through interference lithography: a level set approach. AB - Interference lithography holds the promise of fabricating large-area, defect-free photonic structures on the sub-micrometer scale both rapidly and cheaply. There is a need for a procedure to establish a connection between the structures that are formed and the parameters of the interfering beams. There is also a need to produce self-supporting three-dimensional bicontinuous structures. A generic technique correlating parameters of the interfering beams with the symmetry elements present in the resultant structures by a level-set approach is developed. A particular space group is ensured by equating terms of the intensity equation to a representative level surface of the desired space group. Single- and multiple-exposure techniques are discussed. The beam parameters for certain cubic bicontinuous structures relevant to photonic crystals, viz.,the diamond(D), the simple cubic (P), and the chiral gyroid (G) are derived by utilizing either linear or elliptically polarized light. PMID- 12747445 TI - Probing depth changes following 2 years of periodontal maintenance therapy including adjunctive controlled release of chlorhexidine. AB - BACKGROUND: Multicenter clinical trials have established that the adjunctive use of the subgingival controlled release of chlorhexidine, (CHX chip), significantly reduces probing depth (PD), improves clinical attachment levels, and reduces bleeding on probing compared to scaling and root planing (SRP) alone for periods of up to 9 months. The present report is based on a phase IV clinical trial to examine the adjunctive use of the CHX chip for routine periodontal maintenance therapy (RPMT) over 2 years. METHODS: Eight hundred thirty-five (835) patients were recruited into the study. At baseline a CHX chip was placed in pocket sites with PD > or = 5 mm. The patients were scheduled to receive RPMT at 3-month intervals with repeated CHX chip placement at sites where the PD remained > or = 5 mm. Patients who did not attend the 24-month recall visit or who failed to attend 2 consecutive time frame examinations were excluded from the analyses. RESULTS: The 595 patients included showed a continuous decrease in PD over 2 years of 0.95 mm. After 2 years, 23.2% of patients had at least 2 pockets showing a reduction in PD of 2 mm or more and 58.9% of the sites had been reduced to a PD of < 5 mm. Only 2.9% (n = 57) of the sites showed an increase in PD of > or = 2 mm. Adverse events were mild to moderate in nature and resolved spontaneously without medication. CONCLUSION: The results of this Phase IV or follow-up trial indicate that the adjunctive use of the CHX chip is a clinically safe and effective treatment option for long-term management of chronic periodontitis. PMID- 12747444 TI - Efficacy of controlled-release subgingival chlorhexidine to enhance periodontal regeneration. AB - BACKGROUND: Periodontal regeneration success may be limited by placing bone grafts and membranes in infected sites. The objective of this study was to test the hypothesis that adjunctive subgingival administration of chlorhexidine gelatin bioresorbable chips enhances bone gain when used in conjunction with guided tissue regeneration. METHODS: This was a single center, blinded, 2-arm parallel design study of 44 subjects with one or more sites with probing depth and clinical attachment loss > or = 5 mm following initial therapy and radiographic evidence of bone loss. The patients were randomly assigned to receive either chlorhexidine (CHX) chip or sham chip placement one week prior to regenerative therapy that included graft placement and site coverage with guided tissue membranes. Patients also received CHX or sham chip placement, per their randomization, adjunctively to scaling and root planing or maintenance procedures. Periodontal examinations were completed at baseline (8 weeks prior to surgery); 1 week prior to surgery; and at 3, 6, and 9 months postsurgery. The major outcomes for the study were changes in bone height and bone mass as measured from standardized radiographs used for quantitative digital subtraction radiography over the 11-month study period. RESULTS: Subjects receiving sham chip placement gained a mean bone height of 1.49 +/- 0.22 mm, while patients receiving the CHX chips gained significantly more bone height (3.54 +/- 0.45 mm; P<0.001). Similarly, subjects receiving CHX chips as an adjunct gained significantly more bone mass (5.57 +/- 0.69 mg; P<0.001) than the standard therapy (2.59 +/- 0.34 mg). CONCLUSIONS: These pilot results indicate that locally delivered, controlled release antimicrobial treatment may improve the amount of bone gain during guided tissue regeneration procedures. These data support the evidence that infection control is an important variable in successful regeneration. PMID- 12747446 TI - Permeability and morphology of dentin after erosion induced by acidic drinks. AB - BACKGROUND: The aims of this study were to evaluate 1) the alterations of dentin permeability after single exposure of dentin to several acidic soft drinks with different acid composition; 2) the effectiveness of smear layer on dentin surface to prevent erosion of sound dentin; and 3) the role of brushing procedures. METHODS: Dentin discs from human third molars were prepared. Each disc was treated with 0.5 M neutral EDTA for 5 minutes to remove the smear layer and to calculate the maximum fluid flow rate for each disc (to which an arbitrary value of 100% was assigned) using a pressure apparatus working at 1.0 psi. An homogeneous thin smear layer was then recreated with an abrasive paper under water. The following acidic drinks were applied for 5 minutes onto dentin surface: cola drink (phosphoric acid), orange fruit juice (ascorbic + citric acid), white wine (tartaric acid), vinegar (acetic acid), and mucolytic syrup (benzoic and tartaric acid). Each sample was then brushed for 3 minutes. Finally, each sample was brushed with a toothpaste and, as the final step, etched with phosphoric acid for 1 minute. Permeability was measured after each step. RESULTS: All acidic drinks were able to statistically increase dentin permeability and to open dentinal tubules by removing the smear layer. CONCLUSIONS: The study demonstrated that acidic drinks increased dentin permeability by removing and dissolving the smear layer and smear plugs. The erosion of peritubular dentin and smear plug removal is the main agent responsible for the increase in dentin permeability and probably for clinical dentin hypersensitivity. Brushing procedures reduced dentin permeability, creating a new fine and thin smear layer. Toothpaste may play a protective role in preventing complete smear layer removal and reducing dentin hypersensitivity by producing a new artificial smear layer and deposit inside tubules. The use and the abuse of acidic drinks may damage dentin and increase the risk for dentin hypersensitivity. PMID- 12747447 TI - The effects of cyclosporin on the collagenolytic activity of gingival fibroblasts. AB - BACKGROUND: The immunosuppressive agent cyclosporin is associated with a number of major side-effects including the development of gingival overgrowth. Although the pathogenesis of cyclosporin-induced gingival overgrowth remains unclear, it has been suggested that the finely regulated balance between extracellular matrix synthesis and degradation may be disturbed, resulting in an accumulation of excess connective tissue components within the gingival tissue. The aim of this study was to investigate the effect of cyclosporin on matrix metalloproteinases (MMP)-1 and tissue inhibitors of MMP (TIMP)-1 expression at the mRNA, protein, and enzyme activity levels. METHODS: Gingival fibroblasts were grown to confluence and then cultured in serum-free medium supplemented with cyclosporin over the concentration range of 0 to 2000 ng/ml. MMP-1 and TIMP-1 mRNA levels in cultures were determined by reverse transcription polymerase chain reaction (RT PCR), protein levels in whole conditioned medium were assessed by enzyme-linked immunosorbent assay (ELISA), and collagenolytic activity determined using a 3H acetylated type I collagen degradation assay. Tissue mRNA levels in normal and overgrown gingiva were also determined by RT-PCR. RESULTS: Results indicated that cyclosporin inhibited MMP-1 expression at both the mRNA and protein level in a dose- and time-dependent fashion. The effects on TIMP-1 expression were less clear, cyclosporin inhibiting mRNA expression, but having no effect on TIMP-1 protein levels at any concentration studied. Addition of the drug resulted in reduced levels of collagenolytic activity in the culture medium. MMP-1 mRNA expression was significantly reduced in overgrown compared to normal tissue. CONCLUSIONS: These results add support to the hypothesis that the accumulation of collagen seen in gingival overgrowth can be explained by a cyclosporin-induced inhibition of collagenolytic activity within the gingival tissues. PMID- 12747448 TI - Transforming growth factor-beta 1 expression in the peri-implant soft tissues of healthy and failing dental implants. AB - BACKGROUND: Transforming growth factor-beta (TGF-beta) is composed of a family of multifunctional polypeptide growth factors involved in embryogenesis, inflammation, regulation of immune response, angiogenesis, wound healing, and extracellular matrix formation. TGF-beta1 is the most common isoform found in human tissues. A role of TGF-beta in the pathogenesis of periodontal disease has been suggested. The aim of the present study was a comparative immunohistochemical evaluation of TGF-beta1 in normal keratinized gingiva and in the peri-implant soft tissues surrounding failing non-submerged implants. METHODS: Twenty patients participated in this study. Ten biopsies from healthy keratinized mucosa and 10 biopsies from peri-implant soft tissues surrounding failing implants were obtained (one biopsy per patient). The biopsies were obtained from different patients. RESULTS: In 5 cases of healthy mucosa, the stromal cells were positive between 1 to 5. In 7 cases, the epithelial layers were positive, between 1 and 18 cells. The superficial epithelial layer was negative in all cases. In 9 cases, there was a positivity of the vascular component, between 2 and 16 vessels. In failing implants, the stromal cells were positive in 6 cases, between 1 and 4. In all cases, cells of the epithelial layers were positive, between 15 and 40. The vascular component was positive in all cases, between 12 and 30 vessels. The differences between TGF-beta1 expression in the epithelium around healthy and failing implants were statistically significant (P < 0.0001). The differences between TGF-beta1 expression in the blood vessels in the soft tissues around healthy and failing implants were also statistically significant (P < 0.0001). No statistically significant difference was observed between the 2 groups in the TGF-beta1 expression in the stromal cells (P = 0.88). CONCLUSION: TGF-beta1 may be one of the most important factors in the regulation of the infiltrate, and in the production of tissue repair with a stimulation of fibroblasts and endothelial cells. PMID- 12747449 TI - In vitro evaluation of the mitogenic effect of platelet-derived growth factor-BB on human periodontal ligament cells cultured with various bone allografts. AB - BACKGROUND: Several studies have documented the role of growth factors in periodontal regeneration. It has been shown that platelet-derived growth factor (PDGF) is a potent stimulator of human periodontal ligament (PDL) cells. A variety of bone graft materials are used to treat osseous defects caused by periodontal disease. We evaluated the mitogenic effect of PDGF on human PDL cells cultured with different allografts to determine which of the allografts with or without PDGF promoted periodontal regeneration. METHODS: Two human demineralized freeze-dried allografts of cortical (DFDBA) and cancellous (DFBA) bone and a non demineralized freeze-dried allograft (FBA) from cancellous bone were used alone or supplemented with PDGF-BB. Human PDL cultures were derived from the mid-root of 2 maxillary premolars extracted for orthodontic reasons. Cells were grown separately in 24-well dishes with or without 20 mg of each bone allograft. On day 2 of quiescence, new medium was added with 10 ng/ml of PDGF-BB. DNA synthesis was estimated by measuring [3H] thymidine incorporation to determine the effects of the test agents on cell proliferation. Cells were processed and subjected to scintillation counting after 48 hours of incubation. Counts per minute (cpm/well) were determined for each sample. RESULTS: There was no statistically significant difference (P<0.05) on PDL cell proliferation when the allografts were used alone. PDL cells exhibited significantly greater proliferative responses to the 2 demineralized bone allografts, DFDBA and DFBA, when combined with PDGF-BB. A statistically significant difference on DNA synthesis was noticed when PDGF-BB was added to PDL cells cultured with FBA. PDL cells displayed no significant increase in mitogenic activity when cultured with PDGF-BB alone. CONCLUSIONS: The findings of this study demonstrate the beneficial role of DFDBA, DFBA, and FBA as synergic agents with PDGF-BB to periodontal regeneration. The significant ability of the 2 decalcified bone allografts, DFDBA and DFBA, combined with PDGF to stimulate PDL cell proliferation might be a useful adjunct in the treatment of periodontal defects. PMID- 12747451 TI - Surgical crown lengthening: evaluation of the biological width. AB - BACKGROUND: Previous surgical crown lengthening studies have investigated positional changes of the free gingival margin but not the biological width. Histological studies utilizing animal models have shown that postoperative crestal resorption allowed reestablishment of the biological width. However, very little work has been done in humans. Therefore, the purpose of this study was to evaluate the positional changes of the periodontal tissues, particularly the biological width, following surgical crown lengthening in human subjects. METHODS: Twenty-three (23) patients who needed surgical crown lengthening to gain retention necessary for prosthetic treatment and/or to access caries, tooth fracture, or previous prosthetic margins entered the study. The following parameters were obtained from line angles of treated teeth (teeth requiring surgical crown lengthening) and adjacent teeth with adjacent and non-adjacent sites: plaque and gingival indexes, free gingival margin, probing depth, attachment level, bone level, direct bone level, and biological width. During surgery, the bone level was reduced based on the future prosthetic margin and predetermined biological width; flaps were placed at the bony crest. Patients were examined at baseline and at 3 and 6 months postoperatively. RESULTS: Eighteen patients completed the study. Overall, the amount of bone resected was 1 to 5 mm. At 90% of treated sites, > or = 3 mm of bone was removed. At 3 months, the apical displacement of the free gingival margin at non-adjacent, adjacent, and treated sites was 2.46 +/- 0.25 mm, 2.68 +/- 0.20 mm, and 3.07 +/- 0.16 mm, respectively. There was no significant change in the position of the free gingival margin from 3 to 6 months. The biological width at all sites was smaller at 3 and 6 months compared to baseline (P<0.05) except for the treated sites, which were not significantly different from baseline at 6 months. CONCLUSIONS: During surgical crown lengthening, the bone level was lowered for placement of the prosthetic margin and reestablishment of the biological width. The biological width, at treated sites, was reestablished to its original vertical dimension by 6 months. In addition, a consistent 3 mm gain of coronal tooth structure was observed at the 3- and 6-month examinations. PMID- 12747450 TI - The putative collagen-binding peptide P-15 promotes fibroblast attachment to root shavings but not hydroxyapatite. AB - BACKGROUND: Regenerative periodontal treatment aims to restore the attachment of the periodontal ligament and gingival collagen fibers to both the cementum of the root surface and alveolar bone. Fibroblasts are the predominant cells of the periodontal ligament and gingiva and have important roles in the function and regeneration of the tooth-supporting apparatus. This study investigated whether a putative collagen-based cell-binding peptide (P-15) increases gingival fibroblast attachment to root shavings and bone replacement graft (BRG) materials. METHODS: Gingival and dermal fibroblast attachment to root shavings and BRG materials, and cell proliferation on root shavings and sections were measured fluorometrically. Root shavings and root sections obtained from periodontally healthy teeth were treated with P-15 at 2 concentrations (200 ng/g or 400 ng/g). Citric acid (CA) treated root materials were also compared to untreated root shavings and root sections that served as negative control groups. RESULTS: Attachment of all cells to bone fragments (whether freeze-dried or demineralized) was significantly greater than to hydroxyapatite (HA)-based BRG materials. The addition of P-15 to HA did not significantly increase gingival or dermal fibroblast attachment. At a concentration of 400 ng/g, P-15 significantly increased gingival and dermal fibroblast attachment to root shavings as compared to untreated shavings. Bone fragments, HA-based BRG materials, and untreated root shavings inhibited gingival fibroblast proliferation. Treatment of root sections with P-15 did not have any effect on gingival fibroblast proliferation. CONCLUSIONS: P-15 is a potential alternative to CA for promoting fibroblast attachment to root surfaces. However, P-15 did not enhance fibroblast proliferation on root sections. PMID- 12747452 TI - Mutations of F110 and C126 of the formyl peptide receptor interfere with G protein coupling and chemotaxis. AB - BACKGROUND: Localized aggressive periodontitis (LAgP) is a disease characterized by rapid loss of alveolar bone in teeth of otherwise healthy patients. Neutrophils from LAgP patients have been shown to exhibit diminished chemotaxis and low levels of formyl peptide receptor (FPR) surface expression. A recent study has associated LAgP with 2 polymorphisms in the FPR: 110Phe-->Ser and 126Cys-->Trp. METHODS: We transfected Chinese hamster ovary cells with wtFPR, FPR 110Phe-->Ser, FPR-126Cys-->Trp, or FPR-110Phe-->Ala and determined their surface expression of FPR, their ligand binding affinity, their G-protein coupling, and their chemotaxis toward N-formyl-methionyl-leucyl-phenylalanine (FMLP). RESULTS: FPR-110Phe-->Ser mutants failed to show any significant surface expression or chemotaxis toward FMLP. FPR-126Cys-->Trp mutants exhibited slightly lower than normal binding affinity, markedly lower G-protein coupling response, and markedly lower chemotaxis toward FMLP than that observed with wtFPR. We also analyzed another FPR-Phe110 mutant, FPR-110Phe-->Ala, to ascertain what the effect of mutating this residue might be in a mutant that could be expressed on the cell surface. The FPR-110Phe-->Ala mutant demonstrated markedly lower surface expression, normal ligand binding affinity, markedly lower G-protein coupling, and markedly lower chemotaxis toward FMLP. CONCLUSIONS: Our data substantiate the hypothesis that the chemotactic defects observed in LAgP patients are due at least in part to molecular alterations in the FPR. The FPR-110Phe-->Ser polymorphism appears to be more defective than the FPR-126Cys-->Trp polymorphism, indicating that patients with the former polymorphism might be expected to exhibit a more severe form of aggressive periodontitis. PMID- 12747454 TI - Membrane type (MT) 1-matrix metalloproteinase (MMP) and MMP-2 expression in ligature-induced periodontitis in the rat. AB - BACKGROUND: Matrix metalloproteinases (MMPs) have been shown to be involved in the degradation of the extracellular matrix (ECM). In particular, MMP-2 and MMP-9 (gelatinase A and gelatinase B, respectively) have been identified as the predominant MMPs during periodontitis. Recent studies have indicated that a novel transmembrane MMP, mebrane-type 1 matrix metalloproteinase (MT1-MMP), can activate pro-MMP-2 in tumor metastasis. This study aims to elucidate the presence and localization of MT1-MMP and MMP-2 in periodontitis in a rat model. METHODS: In 2 groups of 40-day-old male Sprague-Dawley rats, periodontitis was initiated by ligating floss around maxillary second molars. A group of control animals were left untreated. Maxillary dentoalveolar segments were isolated after 7 and 21 days postinduction and were prepared for gross and radiographic analysis of bone loss and for histological analysis. Samples were also prepared for gel zymography to detect the presence of MMP-2, and for Northern blot analysis and in situ hybridization with MT1-MMP probes. RESULTS: MMP-2 expression increased at 21 days following ligature placement, in conjunction with MT1-MMP expression. MT1-MMP mRNA expression was observed in epithelial cells, fibroblasts, and in multinucleated cells in the periodontium. CONCLUSION: Our data suggest that MT1 MMP may play a role in extracellular matrix degradation during periodontitis, in concert with MMP-2 and other proteinases. PMID- 12747453 TI - The influence of gingival margin recession on loss of clinical attachment in alcohol-dependent patients without medical disorders. AB - BACKGROUND: The objective of this study was to examine the effects of alcohol and cocaine misuse on periodontal status in a group of alcohol-dependent patients. METHODS: Forty verified alcoholics, either exclusively (n = 10) or with cocaine abuse (n = 30), and a matched comparison group of 25 non-alcoholic subjects, 14 of whom abused cocaine, were entered in the study. All subjects were free from systemic illnesses. Blood levels of gamma glutamyl transpeptidase (GGTP), a liver enzyme indicator of alcohol drinking, were determined. A comprehensive periodontal examination was performed on 6 sites per tooth. The gingival index (GI) and plaque index (PI) were recorded. Attachment levels (AL) were computed as probing depth (PD) plus gingival margin level (GM). RESULTS: No statistically significant differences were noted between the groups for average AL, PD, GM, GI, and PI. In alcoholics, Pearson correlation showed a positive association between GGTP levels and loss of periodontal attachment (P<0.05). A series of regression analyses predicting AL from selected periodontal and demographic factors showed that alcoholics manifest AL by greater increases in GM than non-alcoholics (P<0.07). Severe alcohol use as measured by GGTP >51 iu/l worsens PI (P<0.07), which adversely impacts GM, GI, PD, and ultimately AL. No significant associations were found between cocaine use and AL. CONCLUSIONS: The results suggest that persistent alcohol abuse increases periodontitis development by heightening the loss of attachment through recession of gingival margins. PMID- 12747456 TI - Evaluation of p53, bcl-2, and interleukin-15 levels in gingival crevicular fluid of cyclosporin A-treated patients. AB - BACKGROUND: Apoptosis plays an important role in the maintenance of tissue homeostasis. Considering that apoptosis mediators may play a role in the pathogenesis of drug-induced gingival overgrowth, this study was conducted to evaluate p53, bcl-2, and interleukin-15 (IL-15) levels in gingival crevicular fluid (GCF) of cyclosporin A (CsA)-treated patients. METHODS: Twenty renal transplant patients exhibiting CsA-induced gingival overgrowth and 15 systemically healthy gingivitis patients were included in the study; 15 systemically and periodontally healthy volunteer subjects served as the healthy control group. GCF samples were obtained from one interdental site with gingival overgrowth (GO+) and one site without (GO-) from each CsA-treated patient; hyperplasia index, probing depth, papilla bleeding index, and plaque presence were recorded. One site from each gingivitis patient and healthy control was selected, GCF samples were obtained, and the same clinical parameters were recorded. GCF p53, bcl-2, and IL-15 levels were analyzed by enzyme-linked immunosorbent assay. The results were tested statistically. RESULTS: p53 and bcl 2 levels were below the minimum detectable level in all GCF samples analyzed. CsA GO+ and CsA GO- sites, as well as gingivitis sites, exhibited significantly higher GCF levels of IL-15 compared to healthy controls (P<0.05). The difference between CsA GO+ sites and gingivitis sites was not statistically significant, although the total amount of IL-15 in CsA GO+ sites was lower than gingivitis sites (P>0.05). The total amount of IL-15 in CsA GO- sites was significantly lower than gingivitis sites (P<0.05). No significant correlation was found between the clinical parameters and GCF IL-15 levels (P>0.05). CONCLUSIONS: The pathogenesis of CsA-induced gingival overgrowth is multifactorial. The findings of the present study indicate that IL-15 may play a role in the pathogenesis of CsA-induced gingival overgrowth due to its interactions with CsA and its role in apoptosis and inflammation. PMID- 12747455 TI - A pilot study on antiplaque effects of mastic chewing gum in the oral cavity. AB - BACKGROUND: Chemical plaque control is a useful aid in mechanical oral hygiene, and various chemical agents have been evaluated as antiplaque agents. It has been shown that mastic chewing gum has antibacterial effects on Helicobacter pylori. In this study, the antiplaque effect of mastic chewing gum was investigated. METHODS: Twenty dental students who were both systemically and periodontally healthy participated in this study. The effects of mastic gum were assessed from 2 double-blinded, randomized studies. In the first trial, after mechanical toothbrushing, the inhibitory effect of mastic gum on bacteria in saliva following its use was compared to a placebo gum. Saliva samples were collected at the end of 1, 2, 3, and 4 hours; diluted; inoculated onto 10% horse blood chocolate agar plates; and cultured anaerobically at 37 degrees C for 48 hours. The total number of bacterial colonies on each plate was calculated (n = 20). In the second trial, the effects of mastic gum on de novo plaque formation on tooth surfaces and gingival inflammation were evaluated over a 7-day period without mechanical oral hygiene following random use of either mastic or placebo chewing gum. The degree of plaque accumulation and gingival inflammation were compared between the 2 groups (n = 10). RESULTS: The total number of bacterial colonies was significantly reduced during the 4 hours of chewing mastic gum compared to the placebo gum (P < 0.05, Student t test). The mastic group showed a significantly reduced plaque index (2.69 +/- 0.29 versus 3.15 +/- 0.24; P = 0.001, Student t test) and gingival index (0.44 +/- 0.15 versus 0.66 +/- 0.23, P = 0.021, Student t test) compared to the placebo group. CONCLUSION: These results suggest that mastic chewing gum is a useful antiplaque agent in reducing the bacterial growth in saliva and plaque formation on teeth. PMID- 12747457 TI - Reliability of clinical attachment level recordings: effects on prevalence, extent, and severity estimates. AB - BACKGROUND: Although unreliability of periodontal measurements is considered mainly to concern attempts to detect true changes in longitudinal studies, it is also a source of variability that may substantially decrease the precision of estimates obtained in cross-sectional studies. This study assessed the effects of measurement errors on estimates of prevalence, extent, and severity of clinical attachment loss. METHODS: Four examiners performed repeat attachment level recordings in 128, 122, 134, and 133 adolescents, respectively, who participated in a study of clinical attachment loss among 9,162 high school students from Santiago, Chile. A total of 48,954 duplicate recordings were made. Two examiner teams carried out repeat examinations among 292 and 254 subjects, respectively, corresponding to a total number of 51,600 duplicate recordings for interexaminer reliability assessment. RESULTS: A sizeable imprecision was noted regarding prevalence estimates, particularly at the > or = 1 mm level, whereas examiners were reasonably consistent in the assessment of the number of sites with clinical attachment loss. Ninety-five percent of the differences regarding the number of sites with clinical attachment loss > or = 1 mm were in the range from -5 to 5 sites for intraexaminer comparisons, and from -9 to 8 sites for interexaminer comparisons. These differences should be seen in light of the number of sites affected, which ranged from 0 to 85 sites. CONCLUSIONS: Examiner variation may preclude comparisons of prevalence estimates between studies. Valid comparisons may be made between studies of extent estimates provided that uniform parameters are used to express the extent of periodontal breakdown. PMID- 12747458 TI - Microvascular response in the periodontal ligament following mucoperiosteal flap surgery. AB - BACKGROUND: When the mucoperiosteal flap is elevated, the gingivo-periosteal vascular plexus and periodontal ligament (PDL) vascular plexus sever their connection with the circulatory tracts that pass through alveolar bone. We studied the effect exerted on the PDL vascular plexus during restoration of the circulatory tract. METHODS: We performed experimental mucoperiosteal flap surgery in adult beagle dogs. Histological specimens, prepared after injecting India ink into the blood vessels on postoperative days 5, 7, 14, 21, 28, and 42, were examined under a light microscope. In addition, vascular corrosion cast specimens of the PDL, into which acrylic resin was injected, were observed using a scanning electron microscope. RESULTS: On postoperative day 5, the PDL vascular plexus had formed new blood vessels toward the bone side and root side, and bone resorption of the alveolar bone proper had initiated primarily around the opening of the Volkmann's canal. From postoperative day 7 to 14, the PDL vascular plexus formed new vessels on the bone side and root side accompanied by bone resorption of the alveolus, and demonstrated a complicated vascular architecture, which gradually organized and transformed into a mesh structure from postoperative day 21. Osteogenesis was initiated and encircled the newly formed vessels, and the alveolar bone proper recovered to a flat morphology. Judging from the quantity of new vessels and bone resorption, the width of the PDL space seemed to be the greatest on postoperative day 14. CONCLUSIONS: When the mucoperiosteal flap was elevated, active wound healing was activated because of angiogenesis from the PDL, which possesses a microcirculatory system. Moreover, it was suggested that angiogenesis of the PDL vascular plexus and subsequent bone resorption of alveolar bone might temporarily reduce the tooth-supporting function and cause postoperative mobility. PMID- 12747459 TI - The segmental osteotomy in the management of malposed implants: a case report and literature review. AB - BACKGROUND: The segmental osteotomy provides an important treatment approach for the correction of skeletal-dental malocclusions that would otherwise be unmanageable with conventional orthodontic care. The combined use of orthodontics and orthognathic surgery also has been adapted for use in the correction of developmental and traumatic deformities. Severely malpositioned implant fixtures may not permit restorative correction even with custom prosthetic appliances. As an alternative to implant removal, which can be associated with significant alveolar destruction, orthognathic surgery may offer a treatment modality to improve fixture alignment, thereby enabling a more favorable esthetic and functional prosthesis. This paper presents a case report and review of the segmental osteotomy procedure. METHODS: A 32-year-old female with congenitally missing maxillary lateral incisors underwent implant surgery for prosthetic replacement of these teeth. Due to severe labial positioning of the implant fixtures, acceptable prosthetic treatment was not possible. A segmental osteotomy procedure was used to reposition the implants. The implants were splinted using a ridged interim suprastructure during the 4-month healing period. An occlusal appliance also was worn during the initial 8 weeks of this healing period. RESULTS: The surgical procedure permitted intrusion and palatal repositioning of the implants. Healing proceeded with no untoward effects. Clinical findings at second-stage surgery were consistent with complete osseous repair and integration of the fixtures. Successful prosthetic treatment was subsequently completed for the patient. CONCLUSION: The segmental osteotomy may provide an alternative treatment to the removal of severely malposed implants or their compromised prosthetic restoration. PMID- 12747460 TI - Altered healing following mucogingival surgery in a patient with Crohn's disease: a literature review and case report. AB - BACKGROUND: Crohn's disease is a chronic inflammatory bowel disease characterized by uncertainty in etiology and pathogenesis occasionally with manifestations in oral mucous membranes. This report reviews the literature on Crohn's disease and presents a patient with Crohn's disease on continuous anti-inflammatory and immunosuppressive medication who showed adverse healing response following surgical treatment of gingival recession type defects. METHODS: A 28-year-old male in generally good health apart from his bowel disease requested treatment of multiple maxillary gingival recessions due to esthetic concerns and root sensitivity. Following oral hygiene instruction, 3 coronally advanced flap procedures were performed in the maxillary anterior region to cover the defects. In 2 of the surgical areas, the exposed root surfaces were treated by ethylenediaminetetraacetic acid (EDTA) in combination with enamel matrix derivative (EMD) before coronally positioning the buccal flap. Postoperatively, chlorhexidine gluconate was used for oral hygiene control. RESULTS: The first surgical procedure, performed as a coronally advanced flap, showed delayed and altered healing. Two weeks postoperatively, the flapped tissue remained intensely red and swollen. In the following 2 surgical sites where EDTA and EMD were applied the healing was uneventful. Differences in immediate tissue response, however, did not influence the 3-month treatment outcome with respect to root coverage. CONCLUSIONS: Patients with Crohn's disease on recommended systemic medications may show a delayed and altered wound healing indicating that periodontal surgery must be closely monitored. Treatment planning should take into account the potential wound healing promoting effects of enamel matrix derivative as well as adverse healing effects of chlorhexidine gluconate administration. PMID- 12747461 TI - Severe alveolar bone loss and gingival hyperplasia as initial manifestation of Burkitt cell type acute lymphoblastic leukemia. AB - BACKGROUND: The purpose of this case report is to present severe alveolar bone destruction and gingival enlargement as initial manifestation of Burkitt cell type acute lymphoblastic leukemia (ALL-L3) in a 14-year-old boy. METHODS: The patient was referred to the periodontology department with a 4-week history of gingival enlargement and loosening of teeth. The clinical examination revealed gingival enlargement and expansion of alveolar mucosa particularly in molar regions of both jaws. Almost all teeth had deep periodontal pockets and severe mobility. While the radiographs showed severe alveolar bone loss which extended to apical thirds of many teeth, the microbiologic analysis revealed that the patient did not harbor major periodontopathogenic bacteria species. The results of blood tests and bone marrow aspiration were compatible with ALL-L3. RESULTS: Remission-induction treatment with BFM-90 ALL chemotherapy protocol was started; however, the patient died 4 weeks after the diagnosis due to neutropenic sepsis. CONCLUSIONS: Although no biopsy was performed, it is possible that the severe periodontal destruction and gingival enlargement in this case may have been due to the infiltration of leukemic cells in gingiva, periodontal ligament, and alveolar bone. The similarities of these findings with numb chin syndrome (NCS) and Burkitt's lymphoma (BL) are discussed in this report. PMID- 12747462 TI - An unusual oral chronic graft-versus-host disease-like syndrome following a liver transplant. AB - BACKGROUND: Giving the immunosuppressive drug tacrolimus (FK506) to liver transplant patients has helped to considerably reduce oral side effects such as gingival hyperplasia. Patients taking cyclosporin who suffer from gingival hyperplasia are often switched to tacrolimus. METHODS: We present here a pediatric liver transplantation case study. The patient has been followed for 5.5 years. She developed oral lesions that resulted in the immunosuppressive therapy being changed from tacrolimus to cyclosporin. In clinical terms, the atypical pathology consisted of hyperpigmented patches on the gingival margin, the internal surfaces of the cheeks, and the intraoral surfaces of the lips. When located on the lips, the hyperpigmented patches were associated with pruriginous and edematous lesions. RESULTS: Optical and electronic microscopic examinations of a gingival tissue sample revealed the presence of melanin incontinence and lichenoid lesions with degenerated keratinocytes and a mild infiltrate of lymphocytes. This points to a chronic graft-versus-host disease (cGvHD)-like syndrome linked to tacrolimus. This diagnosis was given further credence by improvement in the lesions following the switch to cyclosporin. CONCLUSION: To our knowledge, this is the first reported case of tacrolimus-associated chronic GvHD-like syndrome occurring in the oral mucosa. PMID- 12747463 TI - Dimensions of peri-implant mucosa: an evaluation of maxillary anterior single implants in humans. AB - BACKGROUND: Attempts have been made to evaluate the biologic dimension of osseointegrated implants, however, most are histologic studies in animals, and the effect of soft tissue support from adjacent teeth on the interproximal dimension of the peri-implant mucosa for anterior single implants has not been addressed. This study clinically evaluated the dimensions of the peri-implant mucosa around 2-stage maxillary anterior single implants in humans after 1 year of function. The influence of the peri-implant biotype was also examined. METHODS: Forty-five patients (20 males and 25 females) with a mean age of 47.3 years were included in this study. A total of 45 maxillary anterior single implant crowns with a mean functional time of 32.5 months (range, 12 to 78) were evaluated. The dimensions of peri-implant mucosa were measured by bone sounding using a periodontal probe at the mesial (MI), mid-facial (F), and distal (DI) aspects of the implant restoration and the proximal aspects (MT, DT) of adjacent natural teeth. In addition, the peri-implant biotype was evaluated and categorized as thick or thin. Statistical analysis was performed using an independent t test (P<0.05). RESULTS: The means and standard deviations of the dimensions of peri-implant mucosa at MT, MI, F, DI, and DT were 4.20 +/- 0.77 mm, 6.17 +/- 1.27 mm, 3.63 +/- 0.91 mm, 5.93 +/- 1.21 mm, and 4.20 +/- 0.64 mm, respectively. The dimensions of peri-implant mucosa in the thick biotype were significantly greater than the thin biotype at MT, MI, and DT (P<0.05). CONCLUSIONS: The mean facial dimension of peri-implant mucosa of 2-stage implants is slightly greater than the average dimension of the dentogingival complex. The level of the interproximal papilla of the implant is independent of the proximal bone level next to the implant, but is related to the interproximal bone level next to the adjacent teeth. Greater peri-implant mucosal dimensions were noted in the presence of a thick peri-implant biotype as compared to a thin biotype. PMID- 12747464 TI - Periodontal terminology revisited. AB - The body of knowledge that was the basis of the science and therapy of periodontics can be found in the literature of the 1950s and 60s. This information conveniently was presented in various textbooks. The evolution of research utilizing histochemistry and electron microscopy, along with disciplined clinical studies, has resulted in an exponential increase in information. A by product of this worldwide effort is confusion resulting from the misuse and interchangeable application of terms disregarding the definitions originally accepted. Even with workshops designed in part to arrive at consensus to preclude confusion, the literature is replete with misleading use of nomenclature. The lack of attention to the appropriate use of terms is, at best, confusing to the student, researcher, and clinician. In the worst case, it may result in poor performance on examinations and improper evaluation of techniques and materials. Many periodontal terms are used inappropriately or are not completely understood, probably because these words have not been adequately described in the literature. Some of the more commonly used and confused terms will be discussed using literature to support the definitions. PMID- 12747465 TI - Re: Rationale for use of antibiotics in periodontics. Walker C, Karpinia K (2002;73:1188-1196). PMID- 12747466 TI - Expression of alphav integrin family in gastric carcinomas: increased alphavbeta6 is associated with lymph node metastasis. AB - To investigate the alterations as to integrin expression in human gastric carcinomas, we analyzed the alphav subunit and 5 types of beta subunits using reverse transcription-polymerase chain reaction (RT-PCR) and competitive RT-PCR. The incidence of alphav, beta6 and beta8 expression was significantly higher in carcinoma tissues than in non-neoplastic gastric mucosal tissues (NGMTs). Out of 18 carcinoma cases with coexpression of alphav and beta6 subunits, which was demonstrated by RT-PCR, 17 cases (94%) showed lymph node metastasis (p = 0.0033). This tendency was confirmed by immunohistochemistry; most cases (23/28, 82%) in which alphavbeta6 integrin was immunohistochemically detected showed lymph node metastasis (p = 0.0193). RT-PCR and immunohistochemical studies showed that gastric carcinoma tissues expressed beta5 subunit in all cases. Furthermore, in a quantitative analysis using competitive RT-PCR, the mean level of beta5 expression was approximately 140 times higher in gastric carcinomas than in NGMTs. Most gastric carcinoma cases (27/38, 71%) were immunohistochemically positive for beta8 subunit. These findings suggest that some members of the alphav integrin family (alphavbeta5, alphavbeta6, alphavbeta8) are up-regulated, and that alphavbeta6 integrin may be involved in the lymphatic metastasis of gastric carcinomas. PMID- 12747467 TI - Expression of cytokeratin 7 and 20 in pathological conditions of the bile tract. AB - Expression of cytokeratin 7 (CK7) and cytokeratin 20 (CK20) helps to establish the origin of biliary and metastatic carcinomas. We investigated the expression of CK7 and CK20 in inflammatory, metaplastic and neoplastic conditions of the bile ducts, and evaluated possible relationships between the CK expression pattern and extrahepatic bile duct/gallbladder carcinomas (EBDCs) or intrahepatic bile duct carcinomas (IBDCs). We used immunohistochemistry for the investigation of 48 formalin-fixed, paraffin-embedded specimens grouped as: A) lithiasic or inflamed surgically resected extrahepatic bile ducts/gallbladders: all were CK7+/CK20+; B) percutaneous liver biopsies from patients with chronic hepatitis C primary biliary cirrhosis and primary sclerosing cholangitis: all were CK7+/CK20 ; C) EBDCs: all were CK7+/CK20+, except for two cases which were CK7-/CK20-; D) IBDCs: all were CK7+/CK20-, except for one case showing CK20 positivity. Metaplastic changes were seen only among specimens in groups A and C: in these cases, CK20 was either focally or diffusely expressed. Our study suggests that the expression of cytokeratins under specific stimuli can be different from normal tissues, and that sometimes CK20 expression can be related to and precede the occurrence of metaplastic alterations. PMID- 12747468 TI - CD44 variant 6 in endometrioid carcinoma of the uterus: its expression in the adenocarcinoma component is an independent prognostic marker. AB - The expression of variant isoforms of CD44 (CD44v) correlates with the metastatic potential of various carcinomas. In endometrial cancer, however, the significance of CD44v-expression as a prognostic indicator has not been fully investigated, nor has it been compared with that of p53, estrogen receptor or Ki67. Surgical material consisted of 14 atypical endometrial hyperplasias (AEH) and 163 endometrial carcinomas (EC). Expression of CD44s, v3 and v6 in carcinoma tissue, and other prognostic markers were immunohistochemically evaluated. The expression in the squamous differentiation was strictly excluded for the evaluation of immunohistochemistry, because the significance was different from that in the adenocarcinoma component. CD44s was frequently expressed in AEH and EC. On the other hand, CD44v3- and v6-positivities were rare or nonexistent in AEH, but were observed in 8 and 35% of EC, respectively. CD44v3-expression correlated significantly with histologic grade and lymph node metastasis. However, there was no correlation between CD44v6 expression and any clinicopathologic factor, nor were other prognostic markers expressed. Univariate analysis revealed that each CD44 was a prognostic determinant in the patients with EC. However, employing multivariate analysis, there were only three independent factors: p53 overexpression, CD44v6 expression and myometrial invasion. CD44v6 expression in the adenocarcinoma component may directly affect the behavior of carcinoma and the prognosis of patients with EC. PMID- 12747469 TI - KAI1 expression in thyroid neoplasms: its linkage with clinicopathologic features in papillary carcinoma. AB - KAI1 is a metastasis suppressor gene located on human chromosome 11p11.2. Previous studies have shown that the down-regulation of KAI1 mRNA and decreased expression of its gene product are significantly linked to carcinoma progression, including metastatic ability. In this study, we investigated KAI1 protein expression in thyroid neoplasms. KAI1 overexpression was observed in 64.0% of papillary carcinoma cases, and the incidence was significantly higher than in cases of follicular carcinoma (20.0%) (p = 0.0001). In papillary carcinomas, decreased KAI1 expression was frequently observed in cases invading beyond the thyroid capsule (p = 0.001), as well as in lymph node metastases (p = 0.0047) and poorly differentiated lesions (p = 0.0299). Furthermore, in anaplastic carcinoma, the incidence of KAI1 overexpression was lower than in papillary carcinoma (p < 0.0001), and only 4.2% of the cases overexpressed this gene. These results suggest that KAI1 down-regulation is significantly related to the progression of papillary carcinoma, including lymph node metastasis, and its anaplastic transformation. PMID- 12747470 TI - Expression of parathyroid hormone (PTH)-related peptide (PthrP) and PTH/PTHrP receptor in osteoclast-like giant cells. AB - Osteoclast-like giant cells (OCGC), which resemble osteoclasts at both the morphologic and immunohistochemical levels, develop in neoplastic tissue. In bone marrow, parathyroid hormone (PTH)-related peptide (PTHrP) can induce osteoclast differentiation by stimulating osteoclast progenitors through the PTH/PTHrP receptor (PPR). To evaluate the possible involvement of PTHrP in OCGC formation in tumors, we analyzed both PTHrP and PPR expression by immunohistochemistry in giant cell tumor of bone (GCTB) and anaplastic thyroid cancer (ATC) containing OCGC. In all cases of either GCTB (n = 5) or ATC (n = 4), intense stainingfor PTHrP was found in OCGC, but only faintly in mononuclear cells. PPR expression in OCGC was also demonstrated in 3 cases of GCTB and 2 cases of ATC. Double staining for PPR and proliferating cell nuclear antigen (PCNA) revealed that PPR was mainly expressed by PCNA-negative mononuclear cells and OCGC in these tumors. This suggests that OCGC might be derived from non-proliferating mononuclear cells by PTHrP stimulation via PPR. Furthermore, the profiles of PTHrP and PPR expression in OCGC were compared with those in the neoplastic GC found in malignancy (n = 6), osteoclasts in bone with osteoarthritis (n = 5), reactive GC, including Langhans-type and foreign body-type in pulmonary tuberculosis (n = 8), and ruptured epidermal cyst (n = 14) in order to clarify whether their distribution pattern was unique to OCGC. In all cases of malignancy, expression of both PTHrP and PPR was observed ubiquitously in neoplastic GC and mononuclear cells regardless of PCNA immunoreactivity. In contrast, in osteoclasts and reactive GC, PTHrP immunoreactivity was seen in all cases and in 7 of 22 cases, respectively, but no PPR expression was observed in either. In situ hybridization confirmed PTHrP expression at the transcriptional level in OCGC and neoplastic GC, but not in osteoclasts. Thus, although PTHrP expression was commonly observed in various types of multinucleated giant cells, their immunohistochemical profiles for PPR were distinct. We conclude that PPR might play a role during OCGC formation in GCTB and ATC. PMID- 12747471 TI - Co-expression of parathyroid hormone and chromogranin A in secondary hyperparathyroidism: a functional marker for secretory activity of hyperplastic nodules. AB - The relationship between secretion of parathyroid hormone (PTH) and biologic characteristics, including cell proliferation or monoclonality, is not yet fully understood. To evaluate secretory activity of glands or nodules histopathologically, we focused on the co-expression of chromogranin A (CgA) and parathyroid hormone (PTH) in each gland or nodule. A total of 55 glands from 38 patients with normal parathyroid glands, hyperplastic glands (diffuse and nodular) and primary adenomas were compared. Co-expression of PTH and CgA was decreased to 44.4% in diffuse hyperplastic glands, and to 39.6% in 91 hyperplastic nodules, in contrast to normal glands and primary adenomas that showed constant co-expression of PTH and CgA. Immunohistochemical study of PTH showed a coarse granular pattern predominantly in PTH-positive/CgA-positive nodules, and a dot-like pattern mainly in PTH-positive/CgA-negative nodules. Laser scanning microscopy and immunoelectron microscopy confirmed that a dot-like pattern is based on a positive reaction of PTH at the Golgi apparatus. MIB-1 LI was 12.6 +/- 11.6 in PTH-positive/CgA-positive, and 19.3 +/- 27.3 in PTH positive/CgA-negative nodules. In conclusion, a combination of PTH and CgA could provide more information about the physiologic state of secretory activity of each nodule than does the simple observation of PTH immunoreactivity. PMID- 12747472 TI - Tumor-to-tumor metastasis of renal cell carcinoma into oncocytic carcinoma of the thyroid. Report of a case and review of the literature. AB - A case of a multifocal oncocytic carcinoma of the thyroid with intratumoral metastases of renal cell carcinoma in a 52-year-old male is reported. Thirteen months before the presentation of the thyroid lesion, the patient underwent nephrectomy for a renal cell carcinoma. The thyroid tumor clinically presented as a palpable nodule. Preoperative fine-needle aspiration cytology showed two cell types: oncocytes and multivacuolated clear cells. The cytologic features were interpreted as a coincidence of an oncocytic tumor of the thyroid and metastasis of a clear cell renal carcinoma. The diagnosis was confirmed by histologic and immunohistochemical examinations. Interestingly, except for metastases within multiple foci of the oncocytic carcinoma, there were no metastatic deposits in nontumoral thyroid. Although the occurrence of tumor-to-tumor metastasis in thyroid gland is exceptionally rare, with only 12 such cases reported to date, one should be aware of this phenomenon to avoid an incorrect diagnosis. To the best of our knowledge, this is the first report of a metastasis into tumor of the thyroid gland, with oncocytic carcinoma being the recipient. PMID- 12747473 TI - Primary histiocytic sarcoma of the spleen associated with erythrophagocytic histiocytosis. AB - We report an exceptional case of a histiocytic sarcoma presenting as a primary isolated spleen tumor in a 71-year-old woman. The neoplastic cells in the cords and sinuses of the red pulp formed multiple lobulated tumors, which were detected in vivo by ultrasound scan. The medium cells, large cells and the giant cells expressed CD68, a histiocyte-associated marker, lysozyme and S100 protein. All these cells were negative for B- and T-cell markers, cytokeratins, melanosome markers (HMB45) and CD1a (Langerhans' cells). Many tumor cells displayed strong erythrophagocytosis and sometimes lymphocytophagocytosis. In addition, numerous histiocytes with morphology indistinguishable from reactive macrophages also exhibited a strong erythrophagocytosis, and were found in the tumor as well as in the normal splenic parenchyma. Despite multi-agent chemotherapy, the patient suffered from a relapse in the liver, with a rapid fatal outcome. A literature review showed that such a primary splenic presentation with multiple tumors is rare. In contrast, in systemic malignant histiocytosis, secondary spleen involvement occurs more frequently but with diffuse infiltration. The association with a reactive histiocytosis with erythrophagocytosis corresponds to "histiocytic medullary reticulosis", as previously described by Scott and Robb Smith. PMID- 12747474 TI - Malignant large cell calcifying Sertoli cell tumor of the testis. AB - A 45-year old man presented with a slow-growing, unilateral beige testicular mass, with a diameter of 4 cm. The testosterone, FSH, LH, estradiol and betahCG serum levels were within normal limits, and there were no associated hormonal syndromes. The patient was treated with inguinal orchidectomy. Microscopically, the tumor was composed of nests of cells with large eosinophilic, slightly granular cytoplasm. There was only a mild degree of atypia and no mitotic activity. The tumor extended into the rete testis. There were intratumoral calcifications, and in the vicinity of the tumor, there was intratubular growth. Although this case is histologically similar to the three previously reported cases of clinically benign large cell calcifying Sertoli cell tumor of the testis with rete testis involvement, the current patient developed right sided para aortic lymph node metastases 18 months after the initial diagnosis. PMID- 12747475 TI - Dose response relationships for acute ionizing-radiation lethality. AB - A review and analysis of the dose response relationship for the probability of acute lethality from prompt or short-term exposure to ionizing radiation is presented. The purpose of this analysis is to provide recommendations concerning estimates of casualties expected from radiation accidents, the use of nuclear weapons, or possible terrorist activities. Previous work on acute ionizing radiation-induced lethality risk together with a collection of dose response relationships are analyzed and presented based on historical case data and expert opinion that have evolved from whole-body radiation therapy experience, radiation exposure accidents, nuclear weapon detonations, and animal experimentation. The nature of the data reviewed ranges from direct individual events to those offered according to collective expert opinion and consensus published as journal articles and in various technical documents and reports. The dose response relationships are expressed as two-parameter (median exposure level and slope) probability distribution models as a function of radiation exposure in terms of a free-in-air dose. Twelve different dose response relationships are presented and discussed, including the impact of some medical care. PMID- 12747477 TI - Characterization of the magnetic fields around walk-through and hand-held metal detectors. AB - Magnetic field strength measurements were made around eight hand-held and 10 walk through metal detectors. The method was similar to that used in previous research for Electronic Article Surveillance units except a Cartesian rather than cylindrical coordinate system was used. Special magnetic field probes specifically designed for metal detector measurements were used. A non-metallic positioning apparatus was designed and fabricated. Magnetic field strength measurements were collected on one hand-held metal detector in the laboratory. The remaining data were collected at airport terminals, federal and state government buildings, and a local high school. Walk-through metal detectors had considerably higher magnetic field strengths [up to 299 Am(-1) p-p (3,741 mG)] than hand-held metal detectors [up to 6 Am(-1) p-p (76 mG)]. The frequencies of the magnetic field signal for walk-through detectors were between 0.1 kHz and 3.5 kHz while those for hand-held detectors were between 89 kHz and 133 kHz. Waveforms for all hand-held metal detectors were sinusoidal; those for walk through metal detectors varied with most being saw-toothed or pulsed. Due to their higher field strengths and the pulsed nature of their magnetic fields, walk through metal detectors likely pose a higher risk for medical device electromagnetic interference than do hand-held units. Root mean squared magnetic field strengths were calculated from the peak-to-peak values and compared to occupational and general public exposure limits. None of these limits were exceeded. Measurement repeatability was examined for one hand-held and two walk through metal detectors. For the hand-held metal detector measurements at the location of the maximum magnetic field strength, measurements by three individuals had a repeatability (percent standard deviation) of 5.9%. Limited repeatability data were collected for on-site measurements of walk-through detectors. One unit showed repeatability of 0.1 to 4.5%; a multi-zone unit showed repeatability of 2.7 to 67.5%. PMID- 12747476 TI - Six-year follow-up of an acute 241Am inhalation intake. AB - A 38-y-old Caucasian male who suffered an acute accidental inhalation intake of 6.3 kBq of 241Am was monitored over 2,135 d using periodic in vivo measurements of the activity in the lungs, liver, and skeleton. Lung clearance was described by a two-compartment exponential model with half-times of 110 d and 10,000 d. The observed uptake of 241Am in the liver (72 Bq) and skeleton (170 Bq) was significantly greater than predicted by the ICRP models for liver (5 Bq) and skeleton (8 Bq). The half-time in the liver was approximately 850 d. Estimates of skeletal activity based on head, wrist, and knee counts generally agreed within 25% over the course of the monitoring period. The half-time in the skeleton was approximately 20,000 d. PMID- 12747478 TI - The monetary value of the averted dose for public exposure assessed by the willingness to pay. AB - The monetary value of the unit averted collective dose at an individual reference dose (as alpha(base)-value) and the aversion against the high individual exposure were assessed by the WTP (Willingness To Pay) method. The original questionnaire and methodology were developed by the CEPN, France, for specialists in the nuclear field. Modifications to the questionnaire were introduced in 2000 to take into account the Hungarian aspects. In 2001, the questionnaire was further modified for use with the public in Hungary. The present paper refers to the results from the most recent studies on public exposure. The questionnaire was provided to 118 persons living in four different regions of Hungary, one near the U-mining site, one near a nuclear power plant, and two others far away from nuclear affected sites. Conversion of the questionnaire to be understandable by the public involved intensive modifications both in form and content. Only 83 to 86 respondents provided usable answers to questions related to the monetary value of the averted dose and the aversion coefficient. The alpha(base)-value was determined from the statistical value of life assessed by the willingness to pay of the respondents for the risk of fatal cancer averted. The mathematical form used to assess the aversion coefficient was a power function with respect to the individual dose, as already introduced in our earlier papers. The advantage of the power function is that the aversion coefficient is independent of the individual reference dose. According to the current results, the mean value of the lognormally distributed alpha-value at the individual reference dose (alpha(base)) takes 10,000 US dollars (person Sv)(-1) with large confidence bounds. The alpha(base)-value is about 50% higher than the alpha(base)-value estimated among the radiation specialists in Hungary, but 5-10 times less than the values obtained in highly developed countries like in France. The normally distributed aversion coefficient for the public assessed to 2.51 +/- 0.56 and is 30% higher than the value obtained by the Hungarian specialists in the nuclear field. Due to the high sensitivity of the alpha-value to the variation of individual reference dose (d0) the derivation of a generic d0-value will depend upon the circumstances of optimization. PMID- 12747479 TI - Transuranic isotopes and 90Sr in attic dust in the vicinity of two nuclear establishments in northern Germany. AB - Attic dust was chosen as the test medium in order to search for traces of man made bone seeking alpha and beta emitters. The samples were taken from 5 houses in the community of Elbmarsch situated at the river Elbe, adjacent to the Krummel nuclear power plant and the nuclear research center of Geesthacht. Five houses in other regions of northern Germany were taken as a control. 238Pu, (239,240)Pu, 241Am, and 244Cm were measured by alpha spectrometry after chemical separation. Additionally, 241Pu was measured by liquid scintillation spectrometry, and the fission product 90Sr was measured in a separate investigation. All nuclides except 244Cm showed activities above the detection limit in the Elbmarsch samples and an elevated mean concentration compared to the control. It can be concluded from the activity ratio 241Am/(239,240)Pu that the Elbmarsch contamination cannot be accounted for by the background levels of transuranic nuclides resulting from weapons fallout. The derived release of alpha emitters is assumed to have contributed to the induction of a leukemia cluster in children, which was observed in Elbmarsch between 1990 and 1996. PMID- 12747480 TI - Inter-species extrapolation of skin heating resulting from millimeter wave irradiation: modeling and experimental results. AB - This study reports measurements of the skin surface temperature elevations during localized irradiation (94 GHz) of three species: rat (irradiated on lower abdomen), rhesus monkey (posterior forelimb), and human (posterior forearm). Two exposure conditions were examined: prolonged, low power density microwaves (LPM) and short-term, high power density microwaves (HPM). Temperature histories were compared with calculations from a bio-heat transfer model. The mean peak surface temperature increase was approximately 7.0 degrees C for the short-term HPM exposures for all three species/locations, and 8.5 degrees C (monkey, human) to 10.5 degrees C (rat) for the longer-duration LPM exposures. The HPM temperature histories are in close agreement with a one-dimensional conduction heat transfer model with negligible blood flow. The LPM temperature histories were compared with calculations from the bio-heat model, evaluated for various (constant) blood flow rates. Results suggest a variable blood flow model, reflecting a dynamic thermoregulatory response, may be more suited to describing skin surface temperature response under long-duration MMW irradiation. PMID- 12747481 TI - Predicting instrument detection efficiency when scanning point and small area radiation sources. AB - Accurate quantification of radionuclides detected during a scanning survey relies on an appropriately determined scan efficiency calibration factor (SECF). Traditionally, instrument efficiency is determined with a stationary instrument and a fixed source geometry. However, as is often the case, the instrument is used in a scanning mode where the source to instrument geometry is dynamic during the observation interval. Procedures were developed to determine the SECF for a point source ("hot particle") and a 10 x 10 cm source passing under the centerline of a 12.7 x 7.62 cm NaI(Tl) detector. The procedures were first tested to determine the SECF from a series of static point source measurements using Monte Carlo N-Particle code. These point static efficiency values were then used to predict the SECF for scan speeds ranging from 10 cm s(-1) to 80 cm s(-1) with a simulated instrument set to collect integrated counts for 1 s. The Monte Carlo N-Particle code was then used to directly determine the SECF by simulating a scan of a point source and 10 x 10 cm area source for scan speeds ranging from 10 cm s(-1) to 80 cm s(-1). Comparison with Monte Carlo N-Particle scan simulation showed the accuracy of the SECF prediction procedures to be within +/-5% for both point and area sources. Experimental results further showed the procedures developed to predict the actual SECF for a point and 10 x 10 cm source to be accurate to within +/-10%. Besides the obvious application to determine an SECF for a given scan speed, this method can be used to determine the maximum detector or source velocity for a desired minimum detectable activity. These procedures are effective and can likely be extended to determine an instrument specific SECF for a range of source sizes, scan speeds, and instrument observation intervals. PMID- 12747482 TI - Radiation dose reduction in invasive cardiology by restriction to adequate instead of optimized picture quality. AB - In this study, the cinegraphic image intensifier entrance dose level for coronary angiography was changed in four steps from dose level A (0.041 microGy frame( 1)), allowing high contrast, but coarse mottled background, to level D (0.164 microGy frame(-1)), affording high transparency and sharpness. Using this new approach throughout the course of 404 consecutive cardiac catheterizations, we reduced patient radiation exposures down to 11 to 16% of currently typical values: i.e., mean dose area products of 5.97 Gy cm2 (n = 91), 6.73 (n = 113), 8.11 (n = 91), and 8.90 (n = 109); cinegraphic dose area products of 2.34, 3.64, 4.56, and 5.49; and cinegraphic dose area products frame(-1) of 13.3, 19.8, 27.0, and 30.2 mGy cm2, for levels A, B, C, and D, respectively. The number of cinegraphic frames ranged within 168 to 182 per case. Our results show that during catheterization interventionalists should vary image intensifier entrance dose levels in accordance with documented structure, angulation, and body mass index. With the exception of cases with special requirements, lower dose levels typically guarantee an adequate image quality. PMID- 12747483 TI - Estimating the depth of embedded contaminants from in-situ gamma spectroscopic measurements. AB - The depth of radiological contamination is information required for decontamination and decommissioning of nuclear facilities. This paper proposes and tests a simple technique for measuring the depth of contamination. The method uses measurements with an HPGe detector at two distances (or heights) from the surface on the same side of the medium. The ratio of the photopeak areas easily can be used to yield the depth of an embedded source. The calculational algorithm was verified by simulations with the MCNP4C code and laboratory experiments for a point source and other special sources. Predictions of the depth for shallowly embedded sources have relatively large uncertainties (approximately 18%) but improve as depth increases. This technique provides a very easy way for a user to quickly estimate the depth of the subsurface contamination on site. PMID- 12747484 TI - Effect of soil amendments on radiocesium transfer to alfalfa. AB - Greenhouse experiments were carried out to evaluate the effect of different soil based countermeasures on radiocesium transfer to Medicago saliva (alfalfa) grown on artificially contaminated loamy-clayey soil. Various rates of potassium, ammonium, and Prussian Blue supplements were applied, and the uptake of radiocesium by control and treated alfalfa plants was monitored during four growth periods. Transfer factors ranging between 0.06 and 0.02 were determined for control plants. Application of potassium at rates higher than 0.1 meq per 100 g soil was found to suppress radiocesium uptake, the effect being more pronounced at increasing fertilization rates. On the contrary, soil treatment with ammonium enhanced the bio-accumulation of radiocesium, indicating that Cs+ ions, previously unavailable to plant roots, were released from soil particles. Prussian Blue supplements had practically no effect on soil-to-alfalfa transfer of the radionuclide. PMID- 12747485 TI - The compact disk as radon detector--a laboratory study of the method. AB - The radon absorption ability and the track etch properties of the polycarbonate material of commercial compact disks make them very useful as sensitive retrospective 222Rn detectors. The basic idea is to remove, after exposure, a surface layer that is thicker than the range of the alpha particles of the 222Rn and 220Rn progenies and to count the electrochemically etched tracks at the corresponding depths (>80 microm). The effects on the response due to differences in pressure, temperature, and humidity have been studied experimentally. The effect of the growing of 210Po after long-term exposures was also estimated. The effect of all listed factors except the temperature is either absent or restricted to maximum--about 10% for the very extreme cases. The variation of the response at 83 microm depth over the temperature interval 15-25 degrees C is +/ 12% around the 20 degrees C value. The dependence of the calibration factor on the etched depth beneath the surface was studied at 4 different temperatures within the range expected indoors. The results show that the depth dependence is exponential with the parameters of the exponent also being dependent on the temperature. In practice, using the track density obtained in two or more depths beneath the compact disk's front surface, an a posteriori temperature correction could be made. By this correction it is possible to substantially reduce the bias in the results due to the unknown temperature during exposure. The results imply that by using home stored compact disks long-term retrospective 222Rn measurements could be made with an uncertainty that could be potentially better than 10%. The useful range of the method starts at about 3 Bq m(-3) (for 10 y exposure time) and appears to cover practically the whole range of indoors 222Rn concentrations. PMID- 12747486 TI - Quality factors for alpha particles in the human respiratory tract. AB - Quality factors of alpha particles emitted by radon progeny in the human lung have been calculated by using the formula recommended by the International Commission on Radiological Protection. Calculations have been carried out for different combinations of sources, energies, and targets. The values obtained are between 20 and 26, with an average of about 24. These are comparable to previously published results for the human lung and for the general consideration of alpha particles emitted in tissue. PMID- 12747487 TI - Investigations of enhanced outdoor radon concentration in Johanngeorgenstadt (Saxony). AB - Since the beginning of the nineties, the German Federal Office for Radiation Protection (Bundesamt fur Strahlenschutz, BfS) has performed extensive measurements of long-term radon concentration in areas influenced by mining. In the region of Johanngeorgenstadt (Saxony) enhanced long-term radon concentrations were measured in the surrounding area of a waste rock pile. To find the explanation for the enhanced radon concentrations both short- and long-term investigations of radon exhalation were performed. To gain information about the local distribution of the radon level, the radon concentrations were measured at a height of 15 cm above ground level. The radon exhalation rate was continuously measured at the toe and the plateau of the waste rock pile. It has been found that radon is extensively released through both diurnal and pronounced seasonal variations. The exhalation pattern is governed by convection processes triggered by the temperature gradient between the waste rock pile and the atmosphere. PMID- 12747488 TI - Robert Hooke's model of memory. AB - In 1682 the scientist and inventor Robert Hooke read a lecture to the Royal Society of London, in which he described a mechanistic model of human memory. Yet few psychologists today seem to have heard of Hooke's memory model. The lecture addressed questions of encoding, memory capacity, repetition, retrieval, and forgetting--some of these in a surprisingly modern way. Hooke's model shares several characteristics with the theory of Richard Semon, which came more than 200 years later, but it is more complete. Among the model's interesting properties are that (1) it allows for attention and other top-down influences on encoding; (2) it uses resonance to implement parallel, cue-dependent retrieval; (3) it explains memory for recency; (4) it offers a single-system account of repetition priming; and (5) the power law of forgetting can be derived from the model's assumptions in a straightforward way. PMID- 12747489 TI - A context-dependent representation model for explaining text repetition effects. AB - The purpose of this article is to review abstract and episodic models of text repetition effects, describe the research supporting these types of models, and propose a new model called the context-dependent representation model, which can explain both abstract-like and episodic-like repetition effects. The basic assumptions of the model are that the surface form and textbase are represented in a context-independent manner, and a coherent situation model binds together the surface features and the textbase and leads to context-dependent representation. When the situation model is well developed, it limits repetition benefits to semantically or contextually similar texts. This produces context dependent repetition effects, which are functionally similar to episodic repetition effects. When the situation model is not well developed or not task relevant, repetition benefits are not limited to contextually similar texts. This produces context-independent repetition effects, which are functionally similar to abstract repetition effects. The context-dependent representation model provides a theoretical basis for explaining past research, and it can serve as a guide for future research aimed at understanding text repetition effects as well as text comprehension and memory. PMID- 12747490 TI - Flexibility versus generalizability in model selection. AB - Which quantitative method should be used to choose among competing mathematical models of cognition? Massaro, Cohen, Campbell, and Rodriguez (2001) favor root mean squared deviation (RMSD), choosing the model that provides the best fit to the data. Their simulation results appear to legitimize its use for comparing two models of information integration because it performed just as well as Bayesian model selection (BMS), which had previously been shown by Myung and Pitt (1997) to be a superior alternative selection method because it considers a model's complexity in addition to its fit. In the present study, after contrasting the theoretical approaches to model selection espoused by Massaro et al. and Myung and Pitt, we discuss the cause of the inconsistencies by expanding on the simulations of Massaro et al. Findings demonstrate that the results from model recovery simulations can be misleading if they are not interpreted relative to the data on which they were evaluated, and that BMS is a more robust selection method. PMID- 12747491 TI - Stimulus and response representations underlying orthogonal stimulus-response compatibility effects. AB - One of the most important findings in recent years regarding response selection is that stimulus-response compatibility (SRC) effects occur for situations in which stimulus and response sets vary along orthogonal dimensions. For two-choice tasks, two types of orthogonal SRC effects are found: an overall advantage for the up-right/down-left mapping, and mapping preferences that vary as a function of position of the response apparatus and responding hand. We review evidence regarding the nature of both types of orthogonal SRC effects. Only asymmetric coding accounts have been proposed for the up-right/down-left advantage, and the evidence indicates that this asymmetry is a property of both verbal and spatial codes. Motoric and coding accounts, as well as a hybrid account based on end state comfort, have been proposed for the second type of orthogonal SRC effect. In this case, the effects of response-apparatus position, hand, and hand posture conform more closely to predictions of the asymmetric coding accounts than to those of the motoric accounts. We also evaluate the mechanisms proposed by the alternative accounts in terms of related literature on the properties of spatial and verbal codes. Evidence indicates that spatial information is represented in categorical and coordinate codes, and both categorical spatial codes and verbal codes are asymmetric. Experiments on mental rotation suggest that it is unlikely that the direction of rotation is determined automatically by movement constraints, as the end-state comfort hypothesis suggests. An explanation in terms of salient features and referential coding can accommodate the range of orthogonal SRC effects. PMID- 12747492 TI - List composition and the word length effect in immediate recall: a comparison of localist and globalist assumptions. AB - Lists of short words usually are recalled better than lists of longer words in immediate recall tasks. Such word length effects might be explained by localist accounts, in which the length of each word in a list affects the recall of that word only, or by globalist accounts, in which the lengths of at least some words affect the recall of other words (e.g., Baddeley, 1986). In a recent localist account, Neath and Nairne (1995) proposed that the recall of each word depends on the likelihood that features within the word are contaminated within the memory representation. We tested this by presenting not only homogeneous lists of short and long words, but also mixed lists, and by including articulatory suppression on some trials. The short-word advantage depended on the composition of the list, ruling out a strictly localist approach. There appear to be several globalist influences on recall, including distinctiveness factors as well as phonological storage and articulation. PMID- 12747493 TI - Perceptual organization influences visual working memory. AB - Previous studies have demonstrated that top-down factors can bias the storage of information in visual working memory. However, relatively little is known about the role that bottom-up stimulus characteristics play in visual working memory storage. In the present study, subjects performed a change detection task in which the to-be-remembered objects were organized in accordance with Gestalt grouping principles. When an attention-capturing cue was presented at the location of one object, other objects that were perceptually grouped with the cued object were more likely to be stored in working memory than were objects that were not grouped with the cued object. Thus, objects that are grouped together tend to be stored together, indicating that bottom-up perceptual organization influences the storage of information in visual working memory. PMID- 12747494 TI - On-line syntactic processing under concurrent memory load. AB - Thirty-six university students were tested in a plausibility judgment task using a self-paced listening paradigm under no-interference and two-digit load conditions. Listening times were longer at syntactically more complex portions of syntactically more complex sentences, and greater loads led to increased listening times. However, listening times at syntactically more complex positions in syntactically more complex sentences did not increase more than listening times at comparable positions in syntactically simple sentences under digit load conditions. The results indicate that a concurrent memory load does not reduce the availability of working memory resources used for on-line syntactic processing and, thus, provide evidence that the working memory system used for assigning syntactic structure is separate from that measured by standard working memory tasks. PMID- 12747495 TI - Concurrent task effects on memory retrieval. AB - Previous studies combining continuous free recall with a concurrent task have generally shown that concurrent tasks impose fairly negligible effects on memory retrieval. By contrast, dual-task studies employing either cued recall or semantic retrieval reveal gross memory impairment and suggest that retrieval is delayed by the centrally demanding phase of the concurrent tasks (i.e., response selection). To explore this conflict, subjects performed continuous free recall while carrying out a serial-choice-response time (RT) task, as in the previous free recall studies. Unlike these previous studies, however, the choice-RT task utilized arbitrary stimulus-response mappings in order to increase the proportion of time devoted to the centrally demanding response selection phase. Recall total was reduced significantly, and recall latency was slowed substantially. PMID- 12747496 TI - Out of sight, out of mind: occlusion and the accessibility of information in narrative comprehension. AB - Do readers encode the perceptual perspectives of characters during narrative comprehension? To address this question, we conducted two experiments using stories that sometimes described situations in which certain information was occluded from the protagonists' views. We generated two related hypotheses concerning the potential impact of occlusion events on text representations. One, the event boundary hypothesis, suggested that any salient narrative event would reduce the accessibility of prior story information. The second, the perceptual availability hypothesis, suggested that accessibility would decrease most for information no longer visible to story protagonists. In Experiment 1, the participants were slowest to respond to verification questions that asked about occluded information. In Experiment 2, we demonstrated that this effect did not extend to other, nonoccluded information. These results suggest that readers encode text information from the perceptual perspective of story protagonists. This is consistent with recent perceptual symbol views of language comprehension. PMID- 12747497 TI - Locus of semantic interference in picture-word interference tasks. AB - Picture-word interference studies typically show that semantically related distractor words embedded within a picture slow picture-naming responses, relative to unrelated ones. This semantic interference effect is commonly interpreted as arising from the competition of lexical-semantic (e.g., Schriefers, Meyer, & Levelt, 1990) or lexical-phonological (e.g., Starreveld & La Heij, 1996) codes. The experiment reported here tests a crucial assumption shared by these accounts--namely, that the effect reflects a lexical, rather than a nonverbal, conceptual conflict. Pictures were named while participants attempted to ignore embedded distractors that were in either verbal or pictorial format. The presence of both words and pictures substantially interfered with naming responses, but only words, not pictures, were found to induce semantic interference. These findings support the claim that for semantic interference to arise, both target picture and distractor have to be lexicalized. Consequently, a general conceptual locus of the effect can be excluded, and the claim that semantic interference is based on a lexical conflict is confirmed. PMID- 12747498 TI - Are implicitly activated associates selectively activated? AB - What constitutes a word's associative past? Words differ in how many associates they activate in memory and, following a brief encounter, those with fewer associates are more likely to be recalled in the presence of related cues. The issue addressed in the present article is whether associative set size effects are produced through the selective activation of strong associates or through the activation of both strong and weak associates. The set size of the strongest associates was varied factorially with the set size of the associates of these associates. We assume that associate set size indexes a word's weaker associates. Experiments 1 and 2 showed that recall varied inversely with both target and associate set sizes. Such results held over variations in study time and participant age. Experiment 3 showed that weak associates of the target had a greater effect on recall when there were more connections among the strongest associates in the set. The findings suggest that activation is not strength selective but includes both weak and strong associates. PMID- 12747499 TI - The effect of dehydroepiandrosterone (DHEA) on recognition memory decision processes and discrimination in postmenopausal women. AB - In this article, the theoretical distinction between recognition memory decision and discrimination processes is used to explore the effect of dehydroepiandrosterone (DHEA) in postmenopausal women. DHEA is an adrenal steroid that diminishes with aging. It has enhanced memory in laboratory animals. An 8 week placebo-controlled, double-blind experiment in which 30 women (ages 39-70) received a 50-mg/day oral dose of DHEA for 4 weeks demonstrated that DHEA made subjects more conservative (i.e., less likely to call test items "old") in their recognition memory decisions and enhanced recognition memory discrimination for items presented briefly. The former result may reflect an empirical regularity (Hirshman, 1995) in which recent strong memory experiences make participants more conservative. The latter result may reflect the effect of DHEA on visual perception, with consequent effects on memory. These results suggest the methodological importance of focusing on decision processes when examining the effects of hormones on memory. PMID- 12747500 TI - Recognizing rotated views of objects: interpolation versus generalization by humans and pigeons. AB - Pigeons and humans were trained to discriminate between pictures of three dimensional objects that differed in global shape. Each pair of objects was shown at two orientations that differed by a depth rotation of 90 degrees during training. Pictures of the objects at novel depth rotations were then tested for recognition. The novel test rotations were 30 degrees, 45 degrees, and 90 degrees from the nearest trained orientation and were either interpolated between the trained orientations or extrapolated outside of the training range. For both pigeons and humans, recognition accuracy and/or speed decreased as a function of distance from the nearest trained orientation. However, humans, but not pigeons, were more accurate in recognizing novel interpolated views than novel extrapolated views. The results suggest that pigeons' recognition was based on independent generalization from each training view, whereas humans showed view combination processes that resulted in a benefit for novel views interpolated between the training views. PMID- 12747501 TI - Decision-bound theory and the influence of familiarity. AB - In this article, we derive a nonparametric prediction from decision-bound theory (DBT). The crucial aspect that is tested is whether or not familiarity of a stimulus affects response time in categorization. We show that, for our design, DBT, extended with some reasonable and testable assumptions, predicts no familiarity effect. Our prediction is nonparametric in that, rather than fit a specific instantiation of general DBT, we posit only some general assumptions of this theory and derive the prediction from these assumptions. It is found that familiarity did have a strong impact on response time for at least half of our participants. We suggest that DBT is in itself incomplete and should be extended to account for the full range of available data. PMID- 12747502 TI - It's not just average faces that are attractive: computer-manipulated averageness makes birds, fish, and automobiles attractive. AB - Average faces are attractive. We sought to distinguish whether this preference is an adaptation for finding high-quality mates (the direct selection account) or whether it reflects more general information-processing mechanisms. In three experiments, we examined the attractiveness of birds, fish, and automobiles whose averageness had been manipulated using digital image manipulation techniques common in research on facial attractiveness. Both manipulated averageness and rated averageness were strongly associated with attractiveness in all three stimulus categories. In addition, for birds and fish, but not for automobiles, the correlation between subjective averageness and attractiveness remained significant when the effect of subjective familiarity was partialled out. The results suggest that at least two mechanisms contribute to the attractiveness of average exemplars. One is a general preference for familiar stimuli, which contributes to the appeal of averageness in all three categories. The other is a preference for averageness per se, which was found for birds and fish, but not for automobiles, and may reflect a preference for features signaling genetic quality in living organisms, including conspecifics. PMID- 12747503 TI - MDFT account of decision making under time pressure. AB - In this paper, decision making under time pressure for multiattribute choice alternatives in a risky environment is investigated. A model, multiattribute decision field theory (MDFT), is introduced that describes both the dynamic and the stochastic nature of decision making and accounts for the observed changes in choice probabilities, including preference reversals as a function of time limit. An experiment in which five different time limits were imposed on the decision maker is presented to test the predictions of the model. It is shown that MDFT is able to account for the complex decision behavior observed in the data. Furthermore, MDFT is compared with the predictions of decision field theory (Busemeyer & Townsend, 1993; Roe, Busemeyer, & Townsend, 2001). PMID- 12747504 TI - Decision making under conflict: decision time as a measure of conflict strength. AB - Conflict and choice are closely related in that choice produces conflict and conflict is resolved by making a choice. Although conflict was invoked in psychological approaches to decision making early on (Lewin, 1931/1964), no generally accepted measure of conflict strength has been established (Tversky & Shafir, 1992). The present study introduces a model (multiattribute decision field theory) that predicts a decision time pattern depending on the conflict situation. In a risky decision-making experiment with multiattribute choice alternatives, decision time is investigated as a possible measure of conflict strength. It is shown that the model can be fitted to a complex choice pattern. PMID- 12747505 TI - Judging confidence influences decision processing in comparative judgments. AB - Current theories of confidence in human judgment assume that confidence and the decision it is based on are inextricably tied to the same process (decisional locus theories) or that confidence processing begins only once the primary decision has been completed (postdecisional locus theories). In the absence of auxiliary assumptions, however, neither class of theory permits the judgment of confidence to affect primary decision processing. In the present study, we examined the effect of rendering confidence judgments on the properties of the decision process in a sensory discrimination task. An examination of the properties of the time taken to determine confidence (i.e., the time taken to render the judgment of confidence) revealed clear evidence of postdecisional confidence processing. Concomitantly, the requirement of confidence judgments was found to substantially increase decisional response times, suggesting that some confidence processing occurs during the primary decision process. We discuss the implications of these findings for contemporary models of confidence in human judgment. PMID- 12747506 TI - Syllogistic reasoning time: disconfirmation disconfirmed. AB - Models of deductive reasoning typically assume that reasoners dedicate more logical analysis to unbelievable conclusions than to believable ones (e.g., Evans, Newstead, Allen, & Pollard, 1994; Newstead, Pollard, Evans, & Allen, 1992). When the conclusion is believable, reasoners are assumed to accept it without much further thought, but when it is unbelievable, they are assumed to analyze the conclusion, presumably in an attempt to disconfirm it. This disconfirmation hypothesis leads to two predictions, which were tested in the present experiment: Reasoners should take longer to reason about problems leading to unbelievable conclusions, and reasoners should consider more models or representations of premise information for unbelievable conclusions than for believable ones. Neither prediction was supported by our data. Indeed, we observed that reasoners took significantly longer to reason about believable conclusions than about unbelievable ones and generated the same number of representations regardless of the believability of the premises. We propose a model, based on a modified version of verbal reasoning theory (Polk & Newell, 1995), that does not depend on the disconfirmation assumption. PMID- 12747507 TI - The multifaceted nature of unsupervised category learning. AB - A substantial portion of category-learning research has focused on one learning mode--namely, classification learning (a supervised learning mode). Subsequently, theories of category learning have focused on how the abstract structure of categories (i.e., the co-occurrence patterns of feature values) affects acquisition. Recent work in supervised learning has shown that a learner's interactions with the stimulus set also plays an important role in acquisition. The present study extends this work to unsupervised learning situations involving simple one-dimensional stimuli. The results suggest that categorization performance is a function of both learning mode (i.e., study conditions) and learning problem (i.e., category structure). Unsupervised learning, like supervised learning, appears to be multifaceted, with different learning modes best paired with certain learning problems. PMID- 12747508 TI - Sequential modulations of stimulus-response correspondence effects depend on awareness of response conflict. AB - In two experiments, sequential modulations of prime-target correspondence effects were investigated in a metacontrast paradigm. Primes were either unmasked and thus consciously discriminable, or entirely masked and thus indiscriminable. Mirroring similar findings from Eriksen- and Simon-type tasks, the influence of prime-target correspondence was reduced in trials that followed a noncorresponding prime-target pair, which suggests that prime-induced response activation can be temporarily suppressed after an incompatible trial. This sequential modulation was independent of prime discriminability in the current trial, but it occurred only when the prime, and thus a conflict between the prime induced and the deliberately to-be-selected response, was consciously experienced in the preceding trial. This suggests that the suppression of automatic response priming is not an immediate consequence of response conflict, but an intention mediated strategy. PMID- 12747509 TI - Inducing blindsight in normal observers. AB - We attempted to induce blindsight in normal observers, in an effort to replicate and extend the findings of Kolb and Braun (1995). In that demonstration, observers were able to localize a target in the absence of visual awareness, indicated by the lack of a correlation between localization accuracy and confidence ratings. Replication of this work seemed essential, given the failed attempt by Morgan, Mason, and Solomon (1997). A key aspect of the present work was the use of a pointing response, which is believed to have access to the unconscious representations subserving blindsight. In the critical rivalrous condition, the display consisted of Gabor patterns presented dichoptically with orthogonal orientation in each eye. Binocular summation of left and right images combined to give the appearance of a uniform plaid, camouflaging the texture defined target. Our attempt to demonstrate blindsight in normal observers was unsuccessful, in that the localization accuracy of the texture-defined target and the observers' confidence ratings were positively correlated. Although the replication was unsuccessful, the results are valuable in that they provide closure to this widely publicized but fruitless line of inquiry. PMID- 12747511 TI - On the determinants of surface brightness. AB - The brightness of an achromatic surface with luminance S on an achromatic background with luminance B varies with S, with B, and with the luminance step deltaL at the border of the surface. In agreement with previous findings indicating that the visual system can perform as a photometer, the results of the two experiments reported here show that S and B determined surface brightness independently of deltaL when the surface was adjacent to and when it was separated from the background. This finding suggests that surface brightness depends on the integration of neural signals representing magnitudes of absolute luminance. A weighted-average model of this integration is proposed. PMID- 12747510 TI - Principles of cross-modal competition: evidence from deficits of attention. AB - How does the attentional system coordinate the processing of stimuli presented simultaneously to different sensory modalities? We investigated this question with individuals with neurological damage who suffered from deficits of attention. In these individuals, we examined how the processing of tactile stimuli is affected by the simultaneous presentation of visual or auditory stimuli. The investigation demonstrated that two stimuli from different modalities are in competition when attention is directed to the perceptual attributes of both, but not when attention is directed to the perceptual attributes of one and the semantic attributes of the other. These findings reveal a differentiated attentional system in which competition is modulated by the level of stimulus representation to which attention is directed. PMID- 12747513 TI - Lung function, airway reactivity, and atopy in newly hired female cotton textile workers. AB - To assess changes in lung function and airway reactivity resulting from exposure to cotton dust, and the role of atopic status in these changes, the authors observed a group of 225 newly hired Chinese textile workers for 1 yr. All workers were female, lifelong nonsmokers, and none of them had been exposed previously to cotton or other occupational dust. Atopic status was determined at baseline. Spirometry, response to methacholine challenge, and total serum immunoglobulin E level were examined at baseline and again after subjects began work in the cotton mills. Obvious cross-shift drops in forced expiratory volume in 1 sec (FEV1.0), and declines in forced vital capacity and FEV1.0 over 1 yr, were observed. Atopic workers had a significantly greater acute drop in FEV1.0 than did nonatopic workers. Both atopic and nonatopic workers had slightly increased airway reactivity at 1 yr, compared with baseline values. The results suggest that exposure to cotton dust is responsible for acute and longitudinal declines in lung function, as well as for slightly increased airway reactivity. Atopy may interact with cotton dust to accentuate the acute lung function response. PMID- 12747512 TI - Analysis of group differences in processing speed: Brinley plots, Q-Q plots, and other conspiracies. AB - Researchers in a growing number of areas (including cognitive development, aging, and neuropsychology) use Brinley plots to compare the processing speed of different groups. Ratcliff, Spieler, and McKoon (2000) argued that a Brinley plot is a quantile-quantile (Q-Q) plot and that therefore Brinley plot regression slopes measure standard deviation ratios rather than relative speed of processing. We show that this argument is incorrect. Brinley plots, by definition, are not Q-Q plots; the former are based on unranked data and the latter are based on ranked data. Furthermore, the relationship between standard deviation ratios and slopes is a general property of regression lines and has no implications for the use of Brinley plot regression slopes as processing speed measures. We also show that the relative speed interpretation of Brinley plot slopes is strongly supported by converging evidence from a metaanalysis of visual search, mental rotation, and memory scanning in young and older adults. As to Ratcliff et al.'s hypothesis that age differences in response time are attributable to greater cautiousness on the part of the elderly, rather than true processing speed differences, this hypothesis has been extensively tested in previous studies and found wanting. PMID- 12747514 TI - Changes in somatosensory evoked potentials, lipid peroxidation, and antioxidant enzymes in experimental diabetes: effect of sulfur dioxide. AB - The effect of sulfur dioxide (SO2) on brain antioxidant status, lipid peroxidation, and somatosensory evoked potentials (SEPs) was investigated in diabetic rats. A total of 40 rats were divided into 4 equal groups: control (C), SO2 + C (SO2), diabetic (D), and SO2 + D (DSO2). Experimental diabetes mellitus was induced by i.v. injection of alloxan at a dose of 50 mg/kg body weight. Ten ppm SO2 was administered to the rats in the sulfur dioxide groups (SO2 and DSO2) in an exposure chamber. Exposure occurred 1 hr/day, 7 days/wk, for 6 wk; control rats were exposed to filtered air during the same time periods. Although SO2 exposure markedly increased copper, zinc Superoxide dismutase activity, it significantly decreased glutathione peroxidase activity in both the diabetic and nondiabetic groups, compared with the C group. Brain catalase activity was unaltered; however, brain thiobarbituric acid reactive substances (TBARS) were elevated in all experimental groups with respect to the C group. SEP components P1, N1, P2, and N2 were significantly increased in all experimental groups, compared with the C group, and these components were also prolonged in the DSO2 group with respect to the other groups. The authors' findings suggest that exposure to SO2, because it increases lipid peroxidation, can change antioxidant enzyme activities and affect SEP components in diabetic rats. PMID- 12747515 TI - Toxic effects of chemical mixtures. AB - Exposures to chemical mixtures have reportedly produced unexpected effects. Examination of new case studies, as well as those previously reported, shows that when the human body is exposed to mixtures of chemicals that include lipophilic and hydrophilic species, the lipophiles facilitate the absorption of the hydrophiles at enhanced levels and produce effects that are not expected from an individual chemical. These effects include enhanced acute and chronic responses, low-level concentration response, and unexpected target organ attack. Octanol:water partition coefficients are predictive of relative lipophilicity and hydrophilicity. The findings have implications for safe drinking water standards, air quality standards, safe industrial and environmental exposure levels, product formulation, product labeling, and protocols for toxicity testing of chemical products. PMID- 12747516 TI - Smoking as a determinant of high organochlorine levels in Greenland. AB - The authors investigated the accumulation of organochlorines among smoking and nonsmoking Inuit hunters (n = 48) in Uummanaq, Greenland, a population with high dietary exposure to persistent organic pollutants (POPs). Human plasma organochlorine levels were positively correlated with age, marine diet, and smoking or plasma cotinine in multiple linear-regression models (p < 0.001). Body mass index was inversely correlated with organochlorine accumulation, independent of smoking status. These findings confirm that the source of POPs among the Inuit in Greenland is diet, but smoking is an important determinant of POP bioaccumulation. Smoking cessation may provide a means to lower the body burden of POPs. PMID- 12747517 TI - Pulmonary function in fur-processing workers: a dose-response relationship. AB - Pulmonary function tests were conducted in 212 male workers exposed to fur dust in a fur-processing factory, and in 148 unexposed male workers. The authors used the cumulative dose of dust exposure (mg-yr) as an exposure index to relate to pulmonary function injury, as measured by pulmonary function tests, in exposed workers. The results showed that fur workers had lower percentages of predicted pulmonary function, as measured by forced vital capacity (FVC), forced expiratory volume in 1 sec (FEV1.0), and maximal flow rate of expiration at 50% and 25% of forced vital capacity (V50 and V25, respectively), compared with controls (p < 0.01). As the cumulative dose of fur dust exposure increased, average levels of pulmonary function declined significantly (p < 0.01), and pulmonary function abnormalities (i.e., < 80% of predicted FVC and FEV1.0, or < 70% of predicted V50 and V25) increased significantly (p < 0.05). Multiple-regression results identified fur dust exposure as the leading risk factor associated with the decline in pulmonary function in the exposed group. The results demonstrated a dose-response relationship between fur dust exposure and respiratory system injury, as measured by pulmonary function tests in fur-processing workers. On the basis of this dose-response relationship and the use of lifetables, the authors proposed an exposure limit of 4 mg/m3 for fur dust. PMID- 12747518 TI - PM10 and asthma medication in schoolchildren. AB - The authors found that the use of asthma medication among elementary schoolchildren was associated with particulate pollution (particulate matter < 10 microm in aerodynamic diameter [PM10]) in a locale where PM10 consisted primarily of coarse fraction material derived from road sanding and reentrained volcanic ash. School nurses' records in 12 neighborhood schools located close to an ambient air monitoring station were abstracted, and the numbers of oral and inhaled doses of asthma medication given daily over a period of 2.5 yr were calculated. Time-series regression models, adjusted for autocorrelation, were developed, with temperature, time trend, day of the week, and month as additional variables. Regression models were estimated, and a lagged moving average of PM10 for 7, 14, 21, and 28 days was used. All models showed positive and significant coefficients for PM10 during periods when asthma medication was administered to the schoolchildren. However, the 21-day moving average was the best fit to the model. PMID- 12747519 TI - Dietary predictors of concentrations of polychlorinated biphenyls in breast adipose tissue of women living in Ontario, Canada. AB - The authors evaluated dietary predictors of concentrations of 11 polychlorinated biphenyls (PCBs) in breast adipose tissue from 190 women with benign breast disease who live in Ontario, Canada. Consumption of fish was positively correlated with, and the most consistent dietary predictor of, breast tissue PCB levels. Dietary predictors accounted for < 8% of the variation in breast adipose tissue PCB levels. Age was positively and most strongly associated with tissue PCB levels, perhaps representing long-term dietary intake and the complex interplay of absorption, metabolism, distribution, and excretion. PMID- 12747520 TI - Hearing loss among workers at an oil refinery in Taiwan. AB - In this study, the authors determined the characteristics of noise-induced hearing loss (NIHL) and its association with hearing-loss-related symptoms (HLRSs) among workers at an oil refinery in Taiwan. Workers from the refinery (N = 384) who had chronic noise exposure (range = 73 to 89 dBa; mean = 81.2, standard deviation = 5.4 dBa) were enrolled in the study. Workers chronically exposed to noise levels that were less than 85 dBa experienced slight hearing loss at high frequencies [(3k + 4k + 6k)/3]. Workers with > or = 1 HLRSs had increased hearing loss at low frequencies [(0.5k + 1k + 2k)/3] and high frequencies (odds ratios = 7.2 and 3.7, respectively). An increased hearing threshold shift for high frequencies (3k, 4k, and 6k Hz) was found in workers who had chronic noise exposure for more than 15 yr. These findings indicate that self reported HLRSs provide valuable early warning signs of NIHL. PMID- 12747521 TI - Do physicians correctly estimate radiation risks from medical imaging? AB - Proper use of medical imaging tools requires knowledge of their associated radiation risks, as well as their possible benefits. The authors assessed physicians' knowledge of the radiation risks associated with bone scintigraphy (bone scan) during an annual meeting of the Israeli Orthopedic Society. The mortality risk of radiation-induced carcinoma from bone scan was identified correctly by less than 5% of respondents. The most frequent answer (38.4%) was the option that was least correct. Senior orthopedists estimated lower risks than did residents. Overall, respondents grossly underestimated the potential radiation risk from bone scan. PMID- 12747522 TI - The theoretical status of latent variables. AB - This article examines the theoretical status of latent variables as used in modern test theory models. First, it is argued that a consistent interpretation of such models requires a realist ontology for latent variables. Second, the relation between latent variables and their indicators is discussed. It is maintained that this relation can be interpreted as a causal one but that in measurement models for interindividual differences the relation does not apply to the level of the individual person. To substantiate intraindividual causal conclusions, one must explicitly represent individual level processes in the measurement model. Several research strategies that may be useful in this respect are discussed, and a typology of constructs is proposed on the basis of this analysis. The need to link individual processes to latent variable models for interindividual differences is emphasized. PMID- 12747523 TI - A symbolic-connectionist theory of relational inference and generalization. AB - The authors present a theory of how relational inference and generalization can be accomplished within a cognitive architecture that is psychologically and neurally realistic. Their proposal is a form of symbolic connectionism: a connectionist system based on distributed representations of concept meanings, using temporal synchrony to bind fillers and roles into relational structures. The authors present a specific instantiation of their theory in the form of a computer simulation model, Learning and Inference with Schemas and Analogies (LISA). By using a kind of self-supervised learning, LISA can make specific inferences and form new relational generalizations and can hence acquire new schemas by induction from examples. The authors demonstrate the sufficiency of the model by using it to simulate a body of empirical phenomena concerning analogical inference and relational generalization. PMID- 12747524 TI - Power, approach, and inhibition. AB - This article examines how power influences behavior. Elevated power is associated with increased rewards and freedom and thereby activates approach-related tendencies. Reduced power is associated with increased threat, punishment, and social constraint and thereby activates inhibition-related tendencies. The authors derive predictions from recent theorizing about approach and inhibition and review relevant evidence. Specifically, power is associated with (a) positive affect, (b) attention to rewards, (c) automatic information processing, and (d) disinhibited behavior. In contrast, reduced power is associated with (a) negative affect; (b) attention to threat, punishment, others' interests, and those features of the self that are relevant to others' goals; (c) controlled information processing; and (d) inhibited social behavior. The potential moderators and consequences of these power-related behavioral patterns are discussed. PMID- 12747525 TI - Unpacking the cognitive map: the parallel map theory of hippocampal function. AB - In the parallel map theory, the hippocampus encodes space with 2 mapping systems. The bearing map is constructed primarily in the dentate gyrus from directional cues such as stimulus gradients. The sketch map is constructed within the hippocampus proper from positional cues. The integrated map emerges when data from the bearing and sketch maps are combined. Because the component maps work in parallel, the impairment of one can reveal residual learning by the other. Such parallel function may explain paradoxes of spatial learning, such as learning after partial hippocampal lesions, taxonomic and sex differences in spatial learning, and the function of hippocampal neurogenesis. By integrating evidence from physiology to phylogeny, the parallel map theory offers a unified explanation for hippocampal function. PMID- 12747526 TI - The cognitive and neural architecture of sequence representation. AB - The authors theorize that 2 neurocognitive sequence-learning systems can be distinguished in serial reaction time experiments, one dorsal (parietal and supplementary motor cortex) and the other ventral (temporal and lateral prefrontal cortex). Dorsal system learning is implicit and associates noncategorized stimuli within dimensional modules. Ventral system learning can be implicit or explicit It also allows associating events across dimensions and therefore is the basis of cross-task integration or interference, depending on degree of cross-task correlation of signals. Accordingly, lack of correlation rather than limited capacity is responsible for dual-task effects on learning. The theory is relevant to issues of attentional effects on learning; the representational basis of complex, sequential skills; hippocampal-versus basal ganglia-based learning; procedural versus declarative memory; and implicit versus explicit memory. PMID- 12747527 TI - A mnemonic theory of odor perception. AB - The psychological basis of odor quality is poorly understood. For pragmatic reason, descriptions of odor quality generally rely on profiling odors in terms of what odorants they bring to mind. It is argued here that this reliance on profiling reflects a basic property of odor perception, namely that odor quality depends on the implicit memories that an odorant elicits. This is supported by evidence indicating that odor quality as well as one's ability to discriminate odors is affected by experience. Developmental studies and cross-cultural research also point to this conclusion. In this article, these findings are reviewed and a model that attempts to account for them is proposed. Finally, the model's consistency with both neurophysiological and neuropsychological data is examined. PMID- 12747528 TI - Recasting (the near-miss to) Weber's law. AB - The authors argue mathematically that a common, power-function model of the just noticeable difference in stimulus intensities is logically inconsistent with an exponent other than 1 in those frequent situations in which a particular averaging over experimental conditions has taken place. The authors show that an alternative power-law model, one which does not share this logical inconsistency, provides a good fit to many well-known, psychoacoustic intensity discrimination data. They also show that the exponent in this alternative model must be nonconstant with the discrimination criterion in experiments implementing this averaging of data. PMID- 12747529 TI - Can anthropomorphic analyses of separation cries in other animals inform us about the emotional nature of social loss in humans? Comment on Blumberg and Sokoloff (2001). AB - M. S. Blumberg and G. Sokoloff's (2001) critical analysis has raised doubt whether emotional feelings can be studied in nonhuman animals, and they have reaffirmed the inappropriateness of anthropomorphic reasoning in animal research. They argue that the ultrasonic distress calls of infant rats may be little more than acoustic by-products of bodily adjustments to physiological stressors. This author argues that comparable vocalizations in other species do index separation distress. Considering that there may be deep homologies in the neural systems that govern such emotional processes in many mammalian species, anthropomorphic zoomorphic reasoning may be a viable cross-species research strategy as long as it is limited to neuroscientific contexts that lead to testable predictions in humans and other animals. PMID- 12747532 TI - Extracorporeal LDL-cholesterol elimination in the treatment of severe familial hypercholesterolemia. AB - The extracorporeal elimination of LDL-cholesterol could be performed using the classic non-selective centrifuge or membrane plasmapheresis. The modern methods are more selective and effective. The atherogenic particules are removed from plasma by active colon or capsula. The methods include: cascade filtration, imunoadsorbtion, heparin-induced precipitation of LDL, thermofiltration, dextran induced precipitation of LDL and direct adsorption of lipids (DALI). The regular LDL-apheresis is the life-saving technique in the treatment of homozygous familial hypercholesterolaemia. It is used in heterozygous familial hypercholesterolaemia when the patients do not respond to diet and drugs therapy, too. The regular LDL-apheresis treatment may be followed by the decreased frequency of angina pain episodes, the reduction of ECG changes during the bicycle ergometry and significant disappearance of tendinous xantomas. Some prospective randomised studies has shown even in this group of patients, resistant to conventional treatment, a significant regression of atherosclerotic changes. PMID- 12747533 TI - Pancreatic-polypeptide in the human pancreas: expression and quantitative variation during development and in ductal adenocarcinoma. AB - AIM: To determine the immunoreactivity of pancreatic-polypeptide (PP) during the development of the human fetal pancreas and ductal pancreatic adenocarcinoma, given that, PP positive cells were demonstrated either into its embryonic anlage or into pancreatic cancer. METHODS: Tissue sections from 15 pancreatic fetal specimens, and equal number of ductal adenocarcinoma specimens, were assessed. RESULTS: The density of positive cells in the primitive exocrine ductal epithelium and endocrine epithelium was significantly higher than the relevant density in the neoplastic pancreatic tissue of mixed (ductal - endocrine) and pure ductal type (p1 = 0.001, p2 < 0.0005, p3 = 0.046 and p4 < 0.0005 respectively). The above values were estimated during the 10th to 12th week. There was no significant difference in the density of positive cells in the mantle zone of the islets from the 13th to the 24th week, and the neoplastic tissue of mixed (p5 = 0.11) and pure ductal type (p6 = 0.23). CONCLUSION: The immunostaining for PP identifies a subgroup of pancreatic ductal adenocarcinomas with a neuroendocrine component, initially considered as pure ductal tumors, and mixed ductal and neuroendocrine tumors. This pattern of expression in neoplasms recapitulates the normal pattern during the embryonal development of the organ, raising the question of therapeutic efficacy of PP and analogues as potential adjuvant treatment of pancreatic cancer. PMID- 12747535 TI - Can we predict maxillary sinus mucosa perforation? AB - This study was carried out to evaluate the prevalence of sinus mucosa perforation occurred during maxillary sinus mucosa elevation surgery, its relation to objective conditions and to the causative medical history, and its influence on postoperative sinusitis, as well. One hundred and forty-six sinus lift procedures have been evaluated in 118 patients. The prevalence of the sinus mucosa perforation was evaluated and subdivided into four groups according to its size and way of treatment. No relation was observed between the perforation and the presence of sinus septa, smoking, radiographic thickening and cyst-like lesions of the maxillary sinus, and previous sinus allergy (P < 0.05). Despite of high prevalence of the perforation of the mucosa (56.16%), no signs of bone graft infection or maxillary sinusitis were noted in any of our patient. PMID- 12747534 TI - Effect of photorefractive keratectomy and laser in situ keratomileusis in high myopia on logMAR visual acuity and contrast sensitivity. AB - PURPOSE: To compare effect of photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK) on contrast sensitivity (CS) and best corrected visual acuity (BCVA) in high myopia. METHODS: 38 myopes (PRK) and 31 patients (LASIK) were examined before and 1, 3, 6, and 12 months postoperatively. Mean preoperative spherical equivalent was -8.0 +/- 1.7D (PRK) and -9.2 +/- 2.1D (LASIK). CS was tested on a computerized system of the Contrast Sensitivity 8010 Type at 6 spatial frequencies (0.74 and 29.55 c/deg), BCVA was measured on logMAR charts. RESULTS: At 12 months postoperatively, mean spherical equivalent was -0.6 +/- 1.0D (PRK) and -1.0 +/- 0.8D (LASIK). Postoperative values of CS were significantly higher in the PRK group, except for spatial frequencies of 3.69 and 7.39 c/deg up to 3 months postoperatively. The initial significant decrease of BCVA lasted up to 6 months after PRK. In the LASIK group BCVA was not significantly different from its preoperative level at the 3-months follow-up. CONCLUSIONS: The significant improvement of CS after PRK suggest that PRK can improve quality of vision in eyes with high myopia. Although recovery of BCVA after LASIK was faster than after PRK, there may be a persistent decrease in CS. PMID- 12747537 TI - Health risk of occupational exposure in welding processes II. Immunological effects. AB - Many of epidemiological studies have certified the relationship between welding and various forms of health damages. In our study we performed an immunological research within a group of twenty men, working in the risky environment of manufacturing of stainless steel constructions (11 welders and 9 grinders, average age was 31 years, 55% of smokers, average time period in welding occupational exposure was 8 years). The exposed group of men was compared with a group of healthy blood donors, marked as the control group (people with various types of employment, living in same locality as a people from exposed group). People within the control group were not occupationally exposed to harmful chemical compounds (from 30 to 100 men were chosen for the individual immune parameters, average age of the whole group was 38 years, 40% of smokers). When compared with the control group, the exposed group of welders and grinders showed higher level of C3 complement (p < 0.001), orosomucoid (p < 0.05), beta-2 microglobulin (p < 0.001), neopterin (p < 0.001) and all fagocytic cells (p < 0.001). On the contrary, in the group of exposed people decreased values of IgA (p < 0.001), IgG (p < 0.001), IgM (p < 0.001), transferin (p < 0.001), alpha-1 antitrypsin (p < 0.001), alpha-2-macroglobulin (p < 0.001), haptoglobulin (p < 0.001) and ceruloplasmin (p < 0.05) were found. Some of these changes were characteristic for the exposed group. They could be considered as precursors of biological markers of effect for given type of exposure. PMID- 12747538 TI - Chemotherapy in a patient with prior history of idiopathic thrombocytopenic purpura. AB - We described a 67 years old small-cell lung cancer patient with a prior history of idiopathic thrombocytopenic purpura (ITP) who was treated with successful chemotherapy. It is probably safe to administer chemotherapeutic agents for some cancer patients with prior history of ITP, but it is important to prevent or minimize the toxicities of these chemotherapeutic agents. PMID- 12747536 TI - Health risk of occupational exposure in welding processes I. Genotoxic risk. AB - The welding processes belong among the important sources of occupational pollutions. The welding fumes are ranked, according to the classification of IARC (International Agency for Research on Cancer), into the group of 2B. In our study we have performed the investigation of twenty men (exposed group) working in the stainless steel welding industrial processes (11 welders and 9 grinders, average age was 31 years, 55% of smokers, average time period of welding occupational exposure was 8 years). The concentrations of chromium (0.557-16.343 mg/m3) and nickel (0.340-10.129 mg/m3) in occupational atmosphere highly exceeded established values of maximum permitted concentrations (0.1 and 1.0 mg/m3, respectively). The concentrations of manganese did not exceed its permitted values. Total concentrations of 12 polycyclic aromatic hydrocarbons (PAH) in occupational atmosphere varied from 300.9 to 961.2 ng/m3. For purposes of biological monitoring, the levels of chromosomal aberrations were determined in the exposed and control group. Healthy blood donors servedas a control group. People from that group were not occupationally exposed to harmful chemical compounds (20 men, average age was 36 years, 40% of smokers). Increased level of chromosomal aberrations of exposed group brought the evidence about higher genotoxic risk of investigated welding processes. PMID- 12747539 TI - Process integration challenges in biotechnology yesterday, today and tomorrow. PMID- 12747540 TI - Back to basics: thermodynamics in biochemical engineering. AB - Rational and efficient process development in chemical technology always makes heavy use of process analysis in terms of balances, kinetics, and thermodynamics. While the first two of these concepts have been extensively used in biotechnology, it appears that thermodynamics has received relatively little attention from biotechnologists. This state of affairs is one among several reasons why development and design of biotechnological processes is today mostly carried out in an essentially empirical fashion and why bioprocesses are often not as thoroughly optimized as many chemical processes. Since quite a large body of knowledge in the area of bio thermodynamics already existed in the early nineties, the Steering Committee of a European Science Foundation program on Process Integration in Biochemical Engineering identified a need to stimulate a more systematic use of thermodynamics in the area. To this effect, a bianual course for advanced graduate students and researchers was developed. The present contribution uses the course structure to provide an outline of the area and to characterize very briefly the achievements, the challenges, and the research needs in the various sub-topics. PMID- 12747541 TI - Integration of physiology and fluid dynamics. AB - The purpose of strategies for the integration of fluid dynamics and physiology is the development of more reliable simulation tools to accelerate the process of scale-up. The rigorous mathematical modeling of the richly interactive relationship between the dynamic response of biosystems and the physical environment changing in time and space must rest on the link between coupled momentum, energy and mass balances and structured modeling of the biophase. With the exponential increase in massive computer capabilities hard- and software tools became available for simulation strategies based on such holistic integration approaches. The review discusses fundamental aspects of application of computational fluid dynamics (CFD) to three-dimensional two-phase turbulence flow in stirred tank bioreactors. Examples of coupling momentum and material balance equations with simple unstructured kinetic models for the behavior of the biophase are used to illustrate the application of these strategies to the selection of suitable impeller configurations. The examples reviewed in this paper include distribution of carbon and energy source in fed batch cultures as well as dissolved oxygen fields during aerobic fermentations. A more precise forecasting of the impact of the multitude of interactions must, however, rest upon a rigorous understanding of the response of the cell factory to the complex dynamic stimulation due to space- and time-dependent concentration fields. The paper also introduces some ideas for fast and very fast experimental observations of intracellular pool concentrations based on stimulus response methods. These observations finally lead to a more complex integration approach based on the coupling of CFD and structured metabolic models. PMID- 12747542 TI - A 'Fine' chemical industry for life science products: green solutions to chemical challenges. AB - Modern biotechnology, in combination with chemistry and process technology, is crucial for the development of new clean and cost effective manufacturing concepts for fine-chemical, food specialty and pharmaceutical products. The impact of biocatalysis on the fine-chemicals industry is presented, where reduction of process development time, the number of reaction steps and the amount of waste generated per kg of end product are the main targets. Integration of biosynthesis and organic chemistry is seen as a key development. The advances in bioseparation technology need to keep pace with the rate of development of novel bio- or chemocatalytic process routes with revised demands on process technology. The need for novel integrated reactors is also presented. The necessary acceleration of process development and reduction of the time-to-market seem well possible, particularly by integrating high-speed experimental techniques and predictive modelling tools. This is crucial for the development of a more sustainable fine-chemicals industry. The evolution of novel 'green' production routes for semi-synthetic antibiotics (SSAs) that are replacing existing chemical processes serves as a recent and relevant case study of this ongoing integration of disciplines. We will also show some challenges in this specific field. PMID- 12747543 TI - Membrane-assisted extractive bioconversions. AB - This chapter summarizes the use of membrane reactors in extractive bioconversions as process integration systems leading to in situ product recovery. Several membrane reactor configurations are analyzed, taking into account the type of bioconversion, biocatalyst type and location (either in the aqueous phase or in the membrane), membrane chemistry and morphology, solvent (extractant) type and its biocompatibility. Modeling of liquid-liquid extractive membrane bioreactors operation is also analyzed considering kinetics and mass-transfer aspects. The chapter includes examples from the authors' laboratory as well as other published in the field. Both enzyme and whole cell-based bioconversions are considered. Relevant aspects related to the solvent (extractant) toxicity and how the membrane could protect the biocatalytic activity are analyzed. Trends in this field are also given. PMID- 12747544 TI - In situ product removal (ISPR) in whole cell biotechnology during the last twenty years. AB - This review sums up the activity in the field of in situ product removal in whole cell bioprocesses over the last 20 years. It gives a complete summary of ISPR operations with microbial cells and cites a series of interesting ISPR applications in plant and animal cell technology. All the ISPR projects with microbial cells are categorized according to their products, their ISPR techniques, and their applied configurations of the ISPR set-up. Research on ISPR application has primarily increased in the field of microbial production of aromas and organic acids such lactic acid over the last ten years. Apart from the field of de novo formation of bioproducts, ISPR is increasingly applied to microbial bioconversion processes. However, despite of the large number of microbial whole cell ISPR projects (approximately 250), very few processes have been transferred to an industrial scale. The proposed processes have mostly been too complex and consequently not cost effective. Therefore, this review emphasizes that the planning of a successful whole cell ISPR process should not only consider the choice of ISPR technique according to the physicochemical properties of the product, but also the potential configuration of the whole process set-up. Furthermore, additional process aspects, biological and legal constraint need to be considered from the very beginning for the design of an ISPR project. Finally, future trends of new, modified or improved ISPR techniques are given. PMID- 12747545 TI - Molecular evolution of adenoviruses. AB - New advances in the field of genetic characterization of adenoviruses originating from different animal species are summarized. Variations seen in the host range and specificity, pathogenicity, genomic arrangement or gene complement are much wider than expected based on previous studies of human adenoviruses. Several exceptional adenoviruses from the two traditional conventional genera are now removed, and proposed to form at least two new genera. The eventual host origin of the new genera, however, is not clarified. Novel results from the genomic and phylogenetic analyses of adenoviruses originating from lower vertebrate species (including reptiles, amphibians and fish) seem to imply that probably five major clusters of adenoviruses exist corresponding to the five major classes of Vertebrata. Adenoviruses, which are now suspected to have common origin with enterobacterium phages from the family Tectiviridae, are perhaps very ancient indeed, and may have undergone a co-evolution with vertebrate hosts. PMID- 12747546 TI - Novel partner proteins of adenovirus penton. AB - Each of the 12 vertices of the adenovirus virion is made of penton, the complex of two oligomeric proteins: a pentameric penton base anchored in the capsid and an antenna-like trimeric fiber extending outwards. Adenovirus penton plays an essential role in the infection of host cells because it is indispensable for virus attachment and internalization. The initial interactions of penton with the primary and secondary receptors are well described. In contrast with that, the role of the penton components downstream of the initial cell contact is not known. This work shows for the first time that two adenovirus structural proteins, fiber and base, are able to interact intimately with different classes of cellular targets. In the case of penton base, a protein responsible for virus internalization, the partners include three ubiquitin-protein ligases that are involved in protein turnover, cell cycle control and endocytosis. Another base protein partner, BAG3, is involved in controlling Hsc70 chaperone activity. Virus attachment protein, fiber, interacts with many different partners, some of them involved in signal transduction and cell growth. Further work will illustrate the implications of these interactions for both the viral and cellular life cycles. PMID- 12747547 TI - Structural studies on adenoviruses. AB - The adenovirus genome encodes more than 40 proteins, of which 11 combine with the viral DNA to form an icosahedral capsid of approximately 150 MDa molecular weight and approximately 900 A in diameter. This chapter reviews the information that structural biology techniques have provided about the adenovirus proteins and capsid. The structures of two capsid proteins (hexon and fiber) and two non structural polypeptides (DNA-binding protein and protease) have been solved by X ray crystallography. Fiber and its knob have been the focus of the latest structural studies, due to their role in host recognition and consequently in virus targeting for human gene therapy. The current model for the large capsid comes from a combination of electron microscopy and crystallography. The resultant images have revealed a surprising similarity between adenovirus and a bacterial virus, which suggests their common evolutionary origin. PMID- 12747548 TI - The multifunctional role of E1A in the transcriptional regulation of CREB/CBP dependent target genes. AB - Oncoproteins encoded by the early region 1A (E1A) of adenoviruses (Ads) have been shown to be powerful tools to study gene regulatory mechanisms. As E1A proteins lack a sequence-specific DNA-binding activity, they modulate viral and cellular gene expression by interacting directly with a diverse array of cellular factors, among them sequence-specific transcription factors, proteins of the general transcription machinery, co-activators and chromatin-modifying enzymes. By making use of these factors, E1A affects major cellular events such as cell cycle control, differentiation, apoptosis, and oncogenic transformation. In this review we will focus on the interaction of E1A with cellular components involved in the cAMP/PKA signal transduction pathway and we will discuss the consequences of these interactions in respect to the activation of CREB/CBP-dependent target genes. PMID- 12747550 TI - Regulation of adenovirus packaging. AB - The application of fundamental concepts about the packaging of the adenovirus genome has contributed significantly to the development of therapeutic viral vectors for gene therapy. The packaging of adenovirus DNA into virus particles requires a cis-acting domain at the left end of the genome. This region contains a series of repeated sequences, termed A repeats due to their AT-rich character, that direct the packaging process. A repeats are believed to represent the binding sites for viral and cellular factors that mediate viral DNA packaging. This review will focus on fundamental aspects of adenovirus DNA packaging as well as how this information has been used and may be used to augment the selectivity of viral DNA packaging in applications pertaining to gene therapy vectors. PMID- 12747549 TI - Adenovirus DNA replication. AB - Replication of the adenovirus genome is catalysed by adenovirus DNA polymerase in which the adenovirus preterminal protein acts as a protein primer. DNA polymerase and preterminal protein form a heterodimer which, in the presence of the cellular transcription factors NFI/CTFI and NFIII/Oct-1, binds to the origin of DNA replication. DNA replication is initiated by DNA polymerase mediated transfer of dCMP onto preterminal protein. Further DNA synthesis is catalysed by DNA polymerase in a strand displacement mechanism which also requires adenovirus DNA binding protein. Here, we discuss the role of individual proteins in this process as revealed by biochemical analysis, mutagenesis and molecular modelling. PMID- 12747551 TI - Adenovirus DNA replication: protein priming, jumping back and the role of the DNA binding protein DBP. AB - The adenovirus (Ad) genome is a linear double-stranded (ds) molecule containing about 36 kilobase pairs. At each end of the genome an approximately 100 base pair (bp) inverted terminal repeat (ITR) is found, the exact length depending on the serotype. To the 5'-end of each ITR, a 55-kDa terminal protein (TP) is covalently coupled. The Ad DNA replication system was one of the first replication systems that could be reconstituted in vitro (Challberg and Kelly 1979). The system requires three virally encoded proteins: precursor TP (pTP), DNA polymerase (Pol) and the DNA binding protein (DBP). In addition, three stimulating human cellular proteins have been identified. These are the transcription factors NFI (Nagata et al. 1982) and Oct-1 (Pruijn et al. 1986) and the type I topoisomerase NFII (Nagata et al. 1983). Ad DNA replication uses a protein primer for replication initiation. The transition from initiation to elongation is marked by a jumping back mechanism (King and van der Vliet 1994), followed by elongation. In order to elongate DBP is required. In this review we discuss the roles of DBP during initiation and elongation and we relate biochemical data on the jumping back mechanism used by Ad Pol to the recently solved crystal structure of a Pol alpha like replication complex (Franklin et al. 2001). We comment on the conditions and possible functions of jumping back and propose a model to describe the jumping back mechanism. PMID- 12747552 TI - The structure and function of the adenovirus major late promoter. AB - The adenovirus major late promoter (MLP) has played a pre-eminent role in the analysis of transcription initiation in mammalian cells, and is an outstanding example of the ways in which the study of adenovirus has led to fundamental insights into general cellular processes. The aim of this chapter is to give a comprehensive review of the structure and function of this model mammalian promoter. After a brief description of late transcription in the adenovirus replication cycle, the experimental evidence for the current consensus on the genetic structure of the MLP, including a consideration of non-primate adenovirus MLPs, will be reviewed. Next, the functions of the MLP in the viral life cycle will be examined, and some of the problems that remain to be resolved will be addressed. The review ends with some ideas on how the knowledge of the structure and function of the MLP can be used in designing virus vectors for specific experimental purposes. PMID- 12747553 TI - Remodelling of the host cell RNA splicing machinery during an adenovirus infection. AB - Adenovirus makes extensive use of RNA splicing to produce a complex set of spliced mRNAs during virus replication. All transcription units, except pIX and IVa2, encode multiple alternatively spliced mRNAs. The accumulation of viral mRNAs is subjected to a temporal regulation, a mechanism that ensures that proteins that are needed at certain stages of the viral life cycle are produced. The complex interaction between host cell RNA splicing factors and viral regulatory elements has been studied intensely during the last decade. Such studies have begun to produce a picture of how adenovirus remodels the host cell RNA splicing machinery to orchestrate the shift from the early to the late profile of viral mRNA accumulation. Recent progress has to a large extent focused on the mechanisms regulating E1A and L1 alternative splicing. Here we will review the current knowledge of cis-acting sequence element, trans-acting factors and mechanisms controlling E1A and L1 alternative splicing. PMID- 12747554 TI - Regulation of mRNA production by the adenoviral E1B 55-kDa and E4 Orf6 proteins. AB - The E1B 55-kDa and E4 Orf6 proteins of human subgroup C adenoviruses both counter host cell defenses mediated by the cellular p53 protein and regulate viral late gene expression. A complex containing the two proteins has been implicated in induction of selective export of viral late mRNAs from the nucleus to the cytoplasm, with concomitant inhibition of export of the majority of newly synthesized cellular mRNAs. The molecular mechanisms by which these viral proteins subvert cellular pathways of nuclear export are not yet clear. Here, we review recent efforts to identify molecular and biochemical functions of the E1B 55-kDa and E4 Orf6 proteins required for regulation of mRNA export, the several difficulties and discrepancies that have been encountered in studies of these viral proteins, and evidence indicating that the reorganization of the infected cell nucleus and production of viral late mRNA at specific intra-nuclear sites are important determinants of selective mRNA export in infected cells. In our view, it is not yet possible to propose a coherent molecular model for regulation of mRNA export by the E1B 55-kDa and E4 Orf6 proteins. However, it should now be possible to address specific questions about the roles of potentially relevant properties of these viral proteins. PMID- 12747555 TI - Adenovirus interaction with its cellular receptor CAR. AB - Representative adenoviruses from four of the five major virus subgroups have been shown to interact with the 46-kDa coxsackievirus and adenovirus receptor (CAR) that is widely expressed on many human cell types, suggesting that the ability to bind CAR may be a conserved feature of many of the approximately 50 known adenovirus serotypes. Receptor binding is a function of the distal 'knob' domain of the trimeric viral fiber protein. Here we review recent structural characterizations of knob, CAR and knob-CAR complexes, and we discuss how knob architecture may have evolved to accommodate opposing selective pressures to vary antigenic structure while conserving receptor binding specificity. In contrast to the hypervariability of the solvent-exposed surface of knob, the CAR receptor was found to be non-polymorphic. PMID- 12747556 TI - Transactivation of human cdc2 promoter by adenovirus E1A. AB - Expression of the adenovirus oncoprotein E1A 12S induces the heterotrimeric transcription factor, NF-Y. NF-Y binds to the two CCAAT motifs upstream of the transcriptional start site of the human cdc2 promoter and is required for activation of the promoter by E1A 12S in cycling cells. The observations that a number of eukaryotic cell cycle regulatory genes also contain the CCAAT motifs and NF-Y binds to them support the notion that E1A 12S could play an important role in deregulated expression of these genes through activation of NF-Y gene in cycling cells. PMID- 12747557 TI - Nuclear perturbations following adenovirus infection. AB - Adenoviruses are processed and assembled in the nuclei of infected cells and thereby produce significant perturbations to their structure and function. As the complex interactions that occur in the nuclei of uninfected cells are not yet fully understood many of the changes seen on infection have been described mainly in morphological terms. This chapter attempts to place more recent findings into this context and demonstrates that adenoviruses are able to hijack many cellular processes and enzymes to their advantage. In particular, modifications to nuclear PODs and nucleoli have more recently been explored in greater detail. PMID- 12747558 TI - The abortive infection of Syrian hamster cells with human adenovirus type 12. AB - Human adenovirus type 12 (Ad12) induces undifferentiated tumors in newborn Syrian hamsters, and this tumor model has been investigated in detail in our laboratory. One of the characteristics of the Ad12-hamster cell system is a strictly abortive infection cycle. In this chapter, we summarize previous and more recent results of studies on the interaction of Ad12 with the nonpermissive BHK21 hamster cell line. The block of Ad12 replication lies before viral DNA replication and late gene transcription which cannot be detected with the most sensitive techniques. Ad12 adsorption, cellular uptake and transport of the viral DNA to the nucleus are less efficient in the nonpermissive hamster cells than in permissive human cells. However, most of the early functions of the Ad12 genome are expressed in BHK21 cells, though at a low level. In the downstream region, the first exon, of the major late promoter (MLP) of Ad12 DNA, a mitigator element of 33 nucleotide pairs in length has been identified which contributes to the inactivity of the MLP in hamster cells and its markedly decreased activity in human cells. The E1 functions of Ad2 or Ad5 are capable of partly complementing the Ad12 deficiencies in hamster cells in that Ad12 viral DNA replication and late gene transcription can proceed, e.g. in a BHK hamster cell line, BHK297-C131,which carries in an integrated form and constitutively expresses the E1 region of Ad5 DNA. Nevertheless, the late Ad12 mRNAs, which are synthesized in this system with the authentic nucleotide sequence, fail to be translated to structural viral proteins. Hence, infectious virions are not produced in the partly complementing system. Probably there is also a translational block for late Ad12 mRNAs in hamster cells. We have recently shown that the overexpression of the Ad12 preterminal protein (pTP) gene or of the E1A gene facilitates the synthesis of full-length, authentic Ad12 DNA in BHK21 cells infected with Ad12. Apparently the pTP has a hitherto unknown function in eliciting full cycles of Ad12 DNA replication even in nonpermissive BHK21 cells when sufficient levels of Ad12 pTP are produced. We pursue the possibility that the completely abortive infection cycle of Ad12 in hamster cells ensures the survival of Ad12-induced hamster tumor cells which all carry, integrated in their genomes, multiple copies of Ad12 DNA. In this way, the viral genomes are immortalized and expanded in a huge number of tumor cells. PMID- 12747559 TI - Perspectives for anaerobic digestion. AB - The modern society generates large amounts of waste that represent a tremendous threat to the environment and human and animal health. To prevent and control this, a range of different waste treatment and disposal methods are used. The choice of method must always be based on maximum safety, minimum environmental impact and, as far as possible, on valorization of the waste and final recycling of the end products. One of the main trends of today's waste management policies is to reduce the stream of waste going to landfills and to recycle the organic material and the plant nutrients back to the soil. Anaerobic digestion (AD) is one way of achieving this goal and it will furthermore, reduce energy consumption or may even be net energy producing. This chapter aims at provide a basic understanding of the world in which anaerobic digestion is operating today. The newest process developments as well as future perspectives will be discussed. PMID- 12747560 TI - Metabolic interactions between methanogenic consortia and anaerobic respiring bacteria. AB - Most types of anaerobic respiration are able to outcompete methanogenic consortia for common substrates if the respective electron acceptors are present in sufficient amounts. Furthermore, several products or intermediate compounds formed by anaerobic respiring bacteria are toxic to methanogenic consortia. Despite the potentially adverse effects, only few inorganic electron acceptors potentially utilizable for anaerobic respiration have been investigated with respect to negative interactions in anaerobic digesters. In this chapter we review competitive and inhibitory interactions between anaerobic respiring populations and methanogenic consortia in bioreactors. Due to the few studies in anaerobic digesters, many of our discussions are based upon studies of defined cultures or natural ecosystems. PMID- 12747561 TI - Kinetics and modeling of anaerobic digestion process. AB - Anaerobic digestion modeling started in the early 1970s when the need for design and efficient operation of anaerobic systems became evident. At that time not only was the knowledge about the complex process of anaerobic digestion inadequate but also there were computational limitations. Thus, the first models were very simple and consisted of a limited number of equations. During the past thirty years much research has been conducted on the peculiarities of the process and on the factors that influence it on the one hand while an enormous progress took place in computer science on the other. The combination of both parameters resulted in the development of more and more concise and complex models. In this chapter the most important models found in the literature are described starting from the simplest and oldest to the more recent and complex ones. PMID- 12747562 TI - Molecular biology of stress genes in methanogens: potential for bioreactor technology. AB - Many agents of physical, chemical, or biological nature, have the potential for causing cell stress. These agents are called stressors and their effects on cells are due to protein denaturation. Cells, microbes, for instance, perform their physiological functions and survive stress only if they have their proteins in the necessary concentrations and shapes. To be functional a protein shape must conform to a specific three-dimensional arrangement, named the native configuration. When a stressor (e.g., temperature elevation or heat shock, decrease in pH, hypersalinity, heavy metals) hits a microbe, it causes proteins to lose their native configuration, which is to say that stressors cause protein denaturation. The cell mounts an anti-stress response: house-keeping genes are down-regulated and stress genes are activated. Among the latter are the genes that produce the Hsp70(DnaK), Hsp60, and small heat protein (sHsp) families of stress proteins. Hsp70(DnaK) is part of the molecular chaperone machine together with Hsp40(DnaJ) and GrpE, and Hsp60 is a component of the chaperonin complex. Both the chaperone machine and the chaperonins play a crucial role in assisting microbial proteins to reach their native, functional configuration and to regain it when it is partially lost due to stress. Proteins that are denatured beyond repair are degraded by proteases so they do not accumulate and become a burden to the cell. All Archaea studied to date possess chaperonins but only some methanogens have the chaperone machine. A recent genome survey indicates that Archaea do not harbor well conserved equivalents of the co-chaperones trigger factor, Hip, Hop, BAG-1, and NAC, although the data suggest that Archaea have proteins related to Hop and to the NAC alpha subunit whose functions remain to be elucidated. Other anti-stress means involve osmolytes, ion traffic, and formation of multicellular structures. All cellular anti-stress mechanisms depend on genes whose products are directly involved in counteracting the effects of stressors, or are regulators. The latter proteins monitor and modulate gene activity. Biomethanation depends on the concerted action of at least three groups of microbes, the methanogens being one of them. Their anti-stress mechanisms are briefly discussed in this Chapter from the standpoint of their role in biomethanation with emphasis on their potential for optimizing bioreactor performance. Bioreactors usually contain stressors that come with the influent, or are produced during the digestion process. If the stressors reach levels above those that can be dealt with by the anti-stress mechanisms of the microbes in the bioreactor, the microbes will die or at least cease to function. The bioreactor will malfunction and crash. Manipulation of genes involved in the anti-stress response, particularly those pertinent to the synthesis and regulation of the Hsp70(DnaK) and Hsp60 molecular machines, is a promising avenue for improving the capacity of microbes to withstand stress, and thus to continue biomethanation even when the bioreactor is loaded with harsh waste. The engineering of methanogenic consortia with stress-resistant microbes, made on demand for efficient bioprocessing of stressor-containing effluents and wastes, is a tangible possibility for the near future. This promising biotechnological development will soon become a reality due to the advances in the study of the stress response and anti-stress mechanisms at the molecular and genetic levels. PMID- 12747563 TI - Molecular ecology of anaerobic reactor systems. AB - Anaerobic reactor systems are essential for the treatment of solid and liquid wastes and constitute a core facility in many waste treatment plants. Although much is known about the basic metabolism in different types of anaerobic reactors, little is known about the microbes responsible for these processes. Only a few percent of Bacteria and Archaea have so far been isolated, and almost nothing is known about the dynamics and interactions between these and other microorganisms. This lack of knowledge is most clearly exemplified by the sometimes unpredictable and unexplainable failures and malfunctions of anaerobic digesters occasionally experienced, leading to sub-optimal methane production and wastewater treatment. Using a variety of molecular techniques, we are able to determine which microorganisms are active, where they are active, and when they are active, but we still need to determine why and what they are doing. As genetic manipulations of anaerobes have been shown in only a few species permitting in-situ gene expression studies, the only way to elucidate the function of different microbes is to correlate the metabolic capabilities of isolated microbes in pure culture to the abundance of each microbe in anaerobic reactor systems by rRNA probing. This chapter focuses on various molecular techniques employed and problems encountered when elucidating the microbial ecology of anaerobic reactor systems. Methods such as quantitative dot blot/fluorescence in-situ probing using various specific nucleic acid probes are discussed and exemplified by studies of anaerobic granular sludge, biofilm and digester systems. PMID- 12747565 TI - Anaerobic granular sludge and biofilm reactors. AB - The long retention time of the active biomass in the high-rate anaerobic digesters is the key factor for the successful application of the high rate anaerobic wastewater treatment. The long solids retention time is achieved due to the specific reactor configuration and it is enhanced by the immobilization of the biomass, which forms static biofilms, particle-supported biofilms, or granules depending on the reactor's operational conditions. The advantages of the high-rate anaerobic digestion over the conventional aerobic wastewater treatment methods has created a clear trend for the change of the role of the anaerobic digestion in the wastewater treatment plants from a pre-treatment method to the main biological treatment method. The application of staged high-rate anaerobic digesters has shown the larger potential among the recent developments in this direction. The most common high-rate anaerobic treatment systems based on anaerobic granular sludge and biofilm are described in this chapter. Emphasis is given to a) the Up-flow Anaerobic Sludge Blanket (UASB) systems, b) the main characteristics of the anaerobic granular sludge, and c) the factors that control the granulation process. Finally, the most innovative staged anaerobic digesters are also presented. PMID- 12747564 TI - Applications of the anaerobic digestion process. AB - At the start of the new millennium waste management has become a political priority in many countries. One of the main problems today is to cope with an increasing amount of primary waste in an environmentally acceptable way. Biowastes, i.e., municipal, agricultural or industrial organic waste, as well as contaminated soils etc., have traditionally been deposited in landfills or even dumped into the sea or lakes without much environmental concern. In recent times, environmental standards of waste incineration and controlled land filling have gradually improved, and new methods of waste sorting and resource/energy recovery have been developed. Treatment of biowastes by anaerobic digestion processes is in many cases the optimal way to convert organic waste into useful products such as energy (in the form of biogas) and a fertilizer product. Other waste management options, such as land filling and incineration of organic waste has become less desirable, and legislation, both in Europe and elsewhere, tends to favor biological treatment as a way of recycling minerals and nutrients of organic wastes from society back to the food production and supply chain. Removing the relatively wet organic waste from the general waste streams also results in a better calorific value of the remainder for incineration, and a more stable fraction for land filling. PMID- 12747566 TI - Potential for anaerobic conversion of xenobiotics. AB - This review covers the latest research on the anaerobic biodegradation of aromatic xenobiotic compounds, with emphasis on surfactants, polycyclic aromatic hydrocarbons, phthalate esters, polychlorinated biphenyls, halogenated phenols, and pesticides. The versatility of anaerobic reactor systems regarding the treatment of xenobiotics is shown with the focus on the UASB reactor, but the applicability of other reactor designs for treatment of hazardous waste is also included. Bioaugmentation has proved to be a viable technique to enhance a specific activity in anaerobic reactors and recent research on reactor and in situ bioaugmentation is reported. PMID- 12747567 TI - Monitoring and control of anaerobic reactors. AB - The current status in monitoring and control of anaerobic reactors is reviewed. The influence of reactor design and waste composition on the possible monitoring and control schemes is examined. After defining the overall control structure, and possible control objectives, the possible process measurements are reviewed in detail. In the sequel, possible manipulated variables, such as the hydraulic retention time, the organic loading rate, the sludge retention time, temperature, pH and alkalinity are evaluated with respect to the two main reactor types: high rate and low-rate. Finally, the different control approaches that have been used are comprehensively described. These include simple and adaptive controllers, as well as more recent developments such as fuzzy controllers, knowledge-based controllers and controllers based on neural networks. PMID- 12747568 TI - Decision tree SAR models for developmental toxicity based on an FDA/TERIS database. AB - Humans are exposed to thousands of environmental chemicals for which no developmental toxicity information is available. Structure-activity relationships (SARs) are models that could be used to efficiently predict the biological activity of potential developmental toxicants. However, at this time, no adequate SAR models of developmental toxicity are available for risk assessment. In the present study, a new developmental database was compiled by combining toxicity information from the Teratogen Information System (TERIS) and the Food and Drug Administration (FDA) guidelines. We implemented a decision tree modeling procedure, using Classification and Regression Tree software and a model ensemble approach termed bagging. We then assessed the empirical distributions of the prediction accuracy measures of the single and ensemble-based models, achieved by repeating our modeling experiment many times by repeated random partitioning of the working database. The decision tree developmental SAR models exhibited modest prediction accuracy. Bagging tended to enhance the accuracy of prediction. Also, the model ensemble approach reduced the variability of prediction measures compared to the single model approach. Further research with data derived from animal species- and endpoint-specific components of an extended and refined FDA/TERIS database has the potential to derive SAR models that would be useful in the developmental risk assessment of the thousands of untested chemicals. PMID- 12747569 TI - Quantitative structure-property relationships for vapor pressures of polybrominated diphenyl ethers. AB - Based on quantum chemical descriptors, by the use of partial least squares regression, quantitative structure-property relationship models for subcooled liquid vapor pressures (PL) of polybrominated diphenyl ether (PBDE) congeners were developed. The Q2cum value of the optimal model obtained is as high as 0.993, indicating a good predictive ability and robustness of the model. Although disagreements were observed between the predicted log PL values and log PL values of validation set, the model obtained can still be used for estimating PL of other PBDE congeners, considering the fact that accurate PL values for compounds with low volatility are extremely difficult to determine experimentally. Intermolecular dispersive interactions play a leading role in governing the values of PL, followed by electrostatic, dipole-dipole and dipole-induced dipole interactions. Intermolecular dispersive interactions also govern the values of enthalpies of vaporization. PMID- 12747571 TI - Quantum descriptors for predictive toxicology of halogenated aliphatic hydrocarbons. AB - In order to improve Quantitative Structure-Activity Relationships (QSARs) for halogenated aliphatics (HA) and to better understand the biophysical mechanism of toxic response to these ubiquitous chemicals, we employ improved quantum mechanical descriptors to account for HA electrophilicity. We demonstrate that, unlike the lowest unoccupied molecular orbital energy, ELUMO, which was previously used as a descriptor, the electron affinity can be systematically improved by application of higher levels of theory. We also show that employing the reciprocal of ELUMO, which is more consistent with frontier molecular orbital (FMO) theory, improves the correlations with in vitro toxicity data. We offer explanations based on FMO theory for a result from our previous work, in which the LUMO energies of HA anions correlated surprisingly well with in vitro toxicity data. Additional descriptors are also suggested and interpreted in terms of the accepted biophysical mechanism of toxic response to HAs and new QSARs are derived for various chemical categories that compose the data set employed. These alternate descriptors provide important insight and could benefit other classes of compounds where the biophysical mechanism of toxic response involves dissociative attachment. PMID- 12747570 TI - E-state modeling of fish toxicity independent of 3D structure information. AB - Topological structure methods are used to model fish toxicity against three classes of organic chemicals. The models were obtained independent of 3D structure information. Further, no mechanism of partitioning was assumed, thus avoiding the problems associated with selection of partitioning system for computation of log P. QSAR models were developed for a set of 92 compounds, including phenols, anilines and substituted aromatic hydrocarbons, yielding excellent statistics: r2 = 0.87, s = 0.25 and q2 = 0.85 leave-one-out (LOO), that are better than those reported in the literature. The model is based on molecular connectivity valence chi-1 index [1chiv], the atom type E-State indices for chlorine [ST(-Cl)] and for ether oxygen [ST(-O-)], and the maximum hydrogen E State atom value in a molecule [Hmax]. Each of the subgroups was also separately well modeled. The model for the full set is validated through use of external validation test sets and ten-fold cross-validation (repeated three times). The quality of the validation statistics supports the claim that the model may be used for estimation of pLC50 values for similar molecules. Detailed structure interpretation is given for the descriptors in the model. These four structure descriptors encode influence of molecular context of groups as well as counts of those groups, in addition to molecular skeletal structure. PMID- 12747572 TI - Quantitative structure-toxicity relationships using TOPS-MODE. 3. Structural factors influencing the permeability of commercial solvents through living human skin. AB - The permeability of a series of 12 commercial solvents through living human skin were studied by using a topological sub-structural approach (TOPS-MODE). We first analyzed the influence of several physicochemical parameters used in describing the skin permeability of the solvents. No single significant relationship was found between any of these physicochemical properties and the permeability of the solvents. A QSAR model using TOPS-MODE descriptors was obtained and validated. This model accounted for more than 95% of the variance in the experimental permeability of these solvents. Using the derived model, the structural factors responsible for the permeability of this series of solvents through living human skin were identified. Methyl groups bonded to heteroatoms or to CH2 groups resulted in the greatest contributions to skin permeability and these groups were considered to be "permeability enhancers". In contrast, groups of the type X = O (X = S, C) were found to be "permeability inhibitors" because they possessed negative contributions to the logarithm of permeability in all of the studied solvents. Drawing on the idea of permeability "enhancers" and "inhibitors", we hypothesized that the solvents needed to orientate themselves in front of the stratum corneum layer first before penetrating through the skin. PMID- 12747573 TI - In-silico screening of high production volume chemicals for mutagenicity using the MCASE QSAR expert system. AB - Computational screening is suggested as a way to set priorities for further testing of high production volume (HPV) chemicals for mutagenicity and other toxic endpoints. Results are presented for batch screening of 2484 HPV chemicals to predict their mutagenicity in Salmonella typhimurium (Ames test). The chemicals were tested against 15 databases for Salmonella strains TA100, TA1535, TA1537, TA97 and TA98, both with metabolic activation (using rat liver and hamster liver S9 mix test) and without metabolic activation. Of the 2484 chemicals, 1868 are predicted to be completely nonmutagenic in all of the 15 data modules and 39 chemicals were found to contain structural fragments outside the knowledge of the expert system and therefore suggested for further evaluation. The remaining 616 chemicals were found to contain different biophores (structural alerts) believed to be linked to mutagenicity. The chemicals were ranked indescending order according to their predicted mutagenic potential and the first 100 chemicals with highest mutagenicity scores are presented. The screening result offers hope that rapid and inexpensive computational methods can aid in prioritizing the testing of HPV chemicals, save time and animals and help to avoid needless expense. PMID- 12747574 TI - Relative molecular similarity in selected chemical carcinogens and the nucleoside triphosphate chain. AB - Several markers of cell toxicity are useful as screening tests for epigenetic carcinogens. The direct effects of chemicals on ATPase and GTPase function are pertinent to the early stages of carcinogenesis. Interference with triphosphate diphosphate exchange mechanisms may result from the interaction of carcinogens with the substrate triphosphate chain. To investigate this hypothesis, a computational chemistry programme is used in this study to investigate molecular similarity in ATPase inhibitors, carcinogens and tumour promoters, in relation to the nucleoside triphosphate chain. The results show that atoms in the investigated molecular structures superimpose on sets of oxygen atoms in the triphosphate chain with interatomic distances < 0.3A. Relative molecular similarity to the substrate triphosphate chain is discussed in terms of the established inhibitory properties of carcinogens/tumour promoters on ATPase function, the carcinogen/ tumour promoting properties of ATPase inhibitors and the prediction of carcinogenic activity from chemical structure. PMID- 12747575 TI - Morphine and d-amphetamine nullify each others' hypothermic effects in mice. AB - We have examined the effects of the psychostimulant d-amphetamine and the neuroleptic haloperidol on hypothermia induced by intraperitoneal injection of the centrally penetrating opioids morphine, fentanyl and sufentanil and the peripherally acting opioid loperamide. Measuring rectal body temperatures, dose response relationships were established for all compounds. Morphine and sufentanil produced hyperthermia at low doses and dose-related hypothermia at higher doses. Fentanyl and loperamide produced dose-related hypothermia. Fixed doses of each opioid producing significant hypothermia were selected for interaction studies. The psychostimulant d-amphetamine was found t o produce biphasic effects with low doses inducing hypothermia and higher doses inducing hyperthemia. Haloperidol produced dose-related hypothermia. The selected doses of the opioids were then injected followed after 15 min. by injection of hypothermia producing doses of d-amphetamine or haloperidol. Hypothermia induced by morphine, fentanyl and sufentanil was reversed by d-amphetamine whereas loperamide-induced hypothermia was unaffected. Rebound hyperthermia was also measured with fentanyl and sufentanil. Haloperidol increased the hypothermic effects of morphine, fentanyl and sufentanil but not of loperamide. In conclusion, the central stimulating effects of opioids and amphetamine may combine resulting in thermogenesis and reversal of hypothermia. Central mechanisms of opioid-induced hypothermia in mice are influenced by drugs which alter the dopamine system, whereas peripheral mechanisms are unaffected. A possible clinical implication for this dopaminergic interaction may be toxicity associated with hyperpyrexia caused by psychostimulant misuse, which is increasingly occurring concomitantly with abuse of opioids. PMID- 12747576 TI - 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine pretreatment attenuates methamphetamine-induced dopamine toxicity. AB - The effects of pretreatment with MPTP (1-methyl4-phenyl-1,2,3,6 tetrahydropyridine) on the acute and long-term effects of methamphetamine on striatal dopamine were evaluated in BALB/c mice. Four subcutaneous injections of a non-toxic dose of MPTP (8 mg/kg, at 2 hr intervals) were followed three days later by a toxic regimen of methamphetamine (four injections of 4 mg/kg, at 2 hr intervals) and mice were sacrificed immediately or three days later. Control mice received saline in place of the MPTP or methamphetamine and mice were observed for acute changes in body temperature, self-injurious behaviour, and striatal dopamine metabolites, or long-term changes in striatal dopamine levels, tyrosine hydroxylase immunoreactivity and glial fibrillary acidic protein. It was observed that pretreatment with MPTP protected mice against the acute increase in body temperature caused by the methamphetamine but, at the same time, delayed the occurrence of self-injurious behaviour following the repeated injections of methamphetamine. Likewise, pretreatment with MPTP attenuated the long-term depletion of striatal dopamine induced by the methamphetamine as well as the large increase in glial fibrillary acidic protein and the reduction in tyrosine hydroxylase immunoreactivity. The MPTP-treatment itself did not alter any of these neurotoxic markers. Finally, the acute decrease in 3,4-dihydroxyphenyacetic acid levels and increased ratio of 3-methoxytyramine/dopamine observed 60 min. after a single injection of methamphetamine (4 mg/kg) were also attenuated in MPTP-treated mice. These results are discussed in the context of the hypothesis that the low-dose treatment with MPTP may modify exchange diffusion across the striatal cell membrane thereby altering the acute and long-lasting effects of methamphetamine. PMID- 12747577 TI - Inhibition of amiodarone-induced lung fibrosis but not alveolitis by angiotensin system antagonists. AB - Earlier work in this laboratory showed that amiodarone induces apoptosis in alveolar epithelial cells by a mechanism inhibitable by angiotensin system antagonists. A variety of recent studies suggests a critical role for alveolar epithelial cell apoptosis in the pathogenesis of lung fibrosis. On this basis we hypothesized that amiodarone-induced alveolar epithelial cell apoptosis and lung fibrosis in vivo might be inhibitable by the angiotensin converting enzyme inhibitor captopril or the angiotensin receptor antagonist losartan. Amiodarone induced lung fibrosis was induced in male Wistar rats by oral administration over six months. Replicate groups of rats received captopril or losartan in addition to amiodarone. Apoptosis was detected by increased total lung activity of caspase 3 and in situ end labeling (ISEL) of fragmented DNA. Collagen was localized and quantitated by the picrosirius red technique. Alveolar epithelial cell apoptosis was detected in amiodarone-treated animals as early as three weeks after the start of amiodarone administration; by six months exposure, the incidence of alveolar epithelial cell apoptosis was significantly reduced by coadministration of captopril or losartan. Alveolar wall collagen accumulation also was significantly attenuated by captopril (100%) or losartan (74%), but neither agent blunted the accumulation of alveolar macrophages evoked by amiodarone (5.3-fold at 6 months). Lung neutrophil content was unchanged by amiodarone treatment for three weeks or six months. These results indicate that amiodarone induces alveolar epithelial cell apoptosis in vivo that is inhibitable by angiotensin antagonists. They also support the hypothesis that blockade of angiotensin formation or function attenuates amiodarone-induced lung fibrosis irrespective of the severity of alveolitis. PMID- 12747578 TI - Effect of acute and chronic lead exposure on apomorpine-induced sniffing in rats. AB - Sniffing is a behaviour which can be induced by dopamine D1/D2 receptor agonists. In order to test the effect of chronic lead exposure on dopamine receptor subtypes, we studied the effects of acute and chronic lead exposure on sniffing induced by apomorphine, a dopamine receptor agonist. Intraperitoneal injection of the dopaminergic receptor agonist, apomorphine (0.25-1 mg/kg), induced dose dependent the sniffing behaviour in rats. Acute administration of lead acetate (50, 100 and 200 mg/kg) deceased the apomorphine-induced sniffing. Chronic lead (0.25%) exposure also decreased the apomophine response Dopamine D1 or D2 receptor antagonists reduced the apomorphine effect. Lead exposure could not potentiate the blockade induced by the dopamine receptor antagonists. It is concluded that the response of lead is not mediated by alteration of dopamine receptors. PMID- 12747579 TI - Effects of dietary beta-cyclodextrin in hypercholesterolaemic rats. AB - Beta-cyclodextrin is a compound that forms inclusion complexes with a variety of molecules, specially bile acids and sterols. This study examines the effects of beta-cyclodextrin on cholesterol and bile acid metabolism in hypercholesterolaemic rats. Male Wistar rats were divided into 4 groups that received during 7 weeks: control diet, 2% cholesterol diet (A), A+2.5% beta cyclodextrin (B) and A+5% beta-cyclodextrin (C). The cholesterol-rich diet induced hepatomegaly and fatty liver and significantly reduced cholesterol, bile acid and phospholipid secretion. Addition of beta-cyclodextrin normalised biliary lipid secretion. Moreover, when compared to A, beta-cyclodextrin significantly lowered plasma phospholipid concentration (B: -21%; C: -29%) and the liver free/total cholesterol molar ratio (B: -40%; C: -38%), increased bile acid faecal output (B: +17%; C: +62%) and enhanced cholesterol 7alpha-hydroxylase activity (B:+50%; C : +100%)and mRNA levels (B: + 14%; C: +29%). 5% beta-cyclodextrin also reduced plasma triglycerides concentration (-38%). However, ALT and AST activities were significantly increased (B: +140% and +280%; C: +72% and +135%) and there was a high incidence of cell necrosis with portal inflammatory cell infiltration. Addition of beta-cyclodextrin to a cholesterol-rich diet results in a triglyceride-lowering action, enhancement of bile acid synthesis and excretion, and normalization of biliary lipid secretion, but produces a marked hepatotoxic effect. PMID- 12747580 TI - Inactivation of cholinesterase induced by chlorpromazine cation radicals. AB - To clarify the mechanism of the side effect of chlorpromazine, we examined the inactivation of cholinesterase induced by chlorpromazine. Cholinesterase was inactivated and its activity was lost in rat serum during interaction of chlorpromazine with horseradish peroxidase and H2O2. When chlorpromazine was oxidized by horseradish peroxidase and H2O2, the reaction solution colored pink and the visible absorption spectrum was consistent with the absorption spectrum of the chlorpromazine cation radical (CPZ*+). Adding cholinesterase immediately decreased the pink color of CPZ*+, indicating that CPZ*+ directly attacked cholinesterase to cause loss of the enzyme activity. Tryptophan residues in cholinesterase sharply decreased during the interaction of cholinesterase with horseradish peroxidase and H2O2. Presumably, loss of tryptophan residues changed the conformation of the cholinesterase protein and then the activity of the enzyme was lost. Other phenothiazine derivatives, including promethazine, triflupromazine, trifluoperazine, trimeprazine, thioridazine and perphenazine, also inactivated cholinesterase during the oxidation by horseradish peroxidase and H2O2. These results suggest that phenothiazine cation radicals participate in toxicological signs caused by the drugs. PMID- 12747581 TI - Pharmacogenetics and pharmacogenomics in drug discovery and development: an overview. AB - Pharmacogenetics is today widely proclaimed as about to revolutionize the face of medicine. In a more realistic assessment, the implementation of molecular genetics and biology will continue to provide us, as it has done already, with better ways to diagnose and treat diseases, but it will do so at a stepwise and evolutionary pace, based on an improved understanding of the nature of disease, allowing more specific treatments, better risk prediction, and the implementation of preventive strategies. As such, future progress in biomedicine will travel the same well-treaded paths of improved differential diagnosis and risk prediction along which it has advanced for the last decades and centuries. So, while meaningful biomedical research today by and large depends on the use of the newly developed tools of genetics and genomics, and the insights we gain through them, it is unlikely to fundamentally change the direction of medical progress. PMID- 12747582 TI - Regulatory gene mutations affecting apolipoprotein gene expression: functions and regulatory behavior of known genes may guide future pharmacogenomic approaches to therapy. AB - A pharmacogenomic approach to therapy requires systematic knowledge of the regulatory regions of the genes, as well as basic understanding of transcriptional regulatory mechanisms of genes. Using the apolipoprotein (apo) A I/CIII gene cluster as a model system, we have identified by in vitro and in vivo studies the regulatory elements and the factors which control its transcription. Studies in transgenic mice established that the hepatocyte nuclear factor (HNF-4) binding site of the apoCIII enhancer, which controls transcription of both genes, is required for the intestinal expression of apoA-I and apoCIII genes, and enhances synergistically their hepatic transcription in vivo. The three Sp1 sites of the enhancer are also required for the intestinal expression of apoA-land apoCIII genes in vivo, and for the enhancement of the hepatic transcription. The regulation of the apoE/apoCI/apoCIV/apoCII cluster is also cited. It is expected that identification of the regulatory regions of genes will be soon accelerated by the sequencing of several mammalian genomes. The functional analyses of the regulatory domains of genes involved in lipid homeostasis, combined with cross species sequence comparisons in the near future, may identify natural regulatory gene polymorphisms in the general population that will permit rational pharmacogenomic approaches for treatment of dyslipidemias. PMID- 12747583 TI - The proteome: anno Domini 2002. AB - We present some current definitions related to functional and structural proteomics and the human proteome, and we review the following aspects of proteome analysis: Classical 2-D map analysis (isoelectric focusing (IEF) followed by SDS-PAGE); Quantitative proteomics (isotope-coded affinity tag (ICAT), fluorescent stains) and their use in e.g., tumor analysis and identification of new target proteins for drug development; Electrophoretic pre fractionation (how to see the hidden proteome!); Multidimensional separations, such as: (a) coupled size-exclusion and reverse-phase (RP)-HPLC; (b) coupled ion exchange and RP-HPLC; (c) coupled RP-HPLC and RP-HPLC at 25/60 degrees C; (d) coupled RP-HPLC and capillary electrophoresis (CE); (e) metal affinity chromatography coupled with CE; Protein chips. Some general conclusions are drawn on proteome analysis and we end this review by trying to decode the glass ball of the aruspex and answer the question: "Quo vadis, proteome"? PMID- 12747584 TI - Cardiac sodium channel diseases. AB - In the last few years, a very active line of research took place after the first identification of SCN5A mutations associated with an inherited form of cardiac arrhythmias and sudden death, the LQT3 variant of the long QT syndrome. Subsequently, two allelic diseases additional to LQT3 were shown to be due to mutations in the same gene, the Brugada syndrome (BrS) and the Lev-Lenegre syndrome (progressive cardiac conduction defect). Genotype-phenotype correlation and in vitro expression studies provide evidence that structure-function relationships of the SCN5A protein are much more complex than initially anticipated. The biophysical characterization of the sodium channel defects associated with different phenotypes and the genotype-phenotype correlation studies brought to the attention of the scientific community a plethora of mechanisms by which even a single amino acid substitution may remarkably affect cardiac excitability. Finally, the evidence of patients harboring an SCN5A mutation and overlapping clinical presentations creates a need for a revision of the traditional classification of the above mentioned diseases. It is now appropriate to consider the "sodium channel syndrome" as a unique clinical entity that may manifest itself with a spectrum of possible phenotypes. PMID- 12747585 TI - Haemophilia B: from molecular diagnosis to gene therapy. AB - Thanks to its typical expression, haemophilia can be identified in writings from the second century AD. Haemophilia B, an X-linked recessive bleeding disorder due to factor IX (FIX) deficiency, has an incidence of about 1:30,000 live male births. The factor 9 (F9) gene was mapped in 1984 on Xq27.1. Haemophilia is diagnosed from prothrombin time, activated partial thromboplastin time, and FIX levels. Carrier females are usually asymptomatic and must be identified only with molecular analysis. Linkage analysis of F9 polymorphisms is rapid and inexpensive but limited by non-informative families, recombinant events, and the high incidence of germline mutations; thus, various procedures have been used for the direct scan of F9 mutations. We set up a novel denaturing high performance liquid chromatographic procedure to scan the F9 gene. This rapid, reproducible procedure detected F9 mutations in 100% of a preliminary cohort of 18 haemophilia B patients. Parallel to the development of more efficient diagnostic tools, the life expectancy and reproductive fitness of haemophilic patients have greatly improved and will continue to improve thanks to the use of less immunogenic recombinant FIX. Hopefully, new approaches based on gene therapy now being evaluated in clinical trials will revolutionise haemophilia B treatment. PMID- 12747586 TI - The use of denaturing high-performance liquid chromatography (DHPLC) for the analysis of genetic variations: impact for diagnostics and pharmacogenetics. AB - Over the past five years, denaturing high-performance liquid chromatography (DHPLC) has emerged as one of the most versatile technologies for the analysis of genetic variations. With the benefit of novel polymer chemistries used for separation, the accuracy, sensitivity, and the throughput of DHPLC for DNA and RNA analysis have greatly improved. DHPLC has been adopted in many laboratories for the screening of mutations and single-nucleotide polymorphisms (SNPs). The ability of DHPLC to detect known and unknown mutations simultaneously has put this technology at the forefront of genetic analysis for a wide variety of diseases. In addition, the high sensitivity of DHPLC combined with the accuracy of the heteroduplex analysis has allowed the development of applications beyond the scope of traditional sequencing or genotyping, e.g., the early detection of cancer. This article reviews the methods, which made DHPLC a widely used tool for diagnosis in molecular genetics and pharmacogenetics. The article provides an overview of current applications in these fields and points to novel applications in areas like epigenetics and the analysis of heteroplasmic mitochondrial DNA, in which DHPLC is becoming the leading technology. PMID- 12747587 TI - Molecular diagnostics by microelectronic microchips. AB - Molecular diagnostics is being revolutionized by the completion of the human genome project and by the development of highly advanced technologies for DNA testing. One of the most important challenges is the introduction of high throughput systems such as DNA chips into diagnostic laboratories. DNA microchips are small devices permitting rapid analysis of genetic information, exploiting miniaturization of all components and automation of operational procedures. The most important biochip applications include gene expression and genetic variation identification and both may improve human molecular diagnostics. Here we review several approaches developed to allow rapid detection of many single nucleotide polymorphisms and mutations in large population samples. Among these, the use of microelectronics seems to best fit with the needs of molecular diagnostics. PMID- 12747588 TI - Molecular beacons as diagnostic tools: technology and applications. AB - Molecular beacons are single-stranded, fluorophore-labeled nucleic acid probes that are capable of generating a fluorescent signal in the presence of target, but are dark in the absence of target. Molecular beacons allow multiplex detection of PCR products in real time in a homogeneous assay format. Real time detection is inherently quantitative and affords a greater dynamic range than end point detection methods. Reactions in a homogeneous assay format are sealed before amplification takes place, providing improved contamination control. A single cycler/reader instrument, coupled with automated sample preparation, results in higher throughput and greater ease of use. A multiplex qualitative assay that detects Chlamydia trachomatis and Neisseria gonorrhoeae, along with an internal control, has been developed. High specificity is achieved through careful selection of primers, probes and assay conditions. Quantitative HIV, HCV, and HBV viral load assays, with sensitivities of 50 copies/ml, 20 IU/ml, and 50 copies/ml, respectively, are achievable. The viral load assays are designed to quantitate all subtype and genotype specimens equivalently. A molecular beacon assay has been designed to detect a single nucleotide polymorphism in the beta2 adrenergic receptor gene. PMID- 12747589 TI - Electrochemical DNA sensor for detection of single nucleotide polymorphisms. AB - In recent years there has been an increased interest in using biosensors for the recognition and monitoring of molecule interactions. DNA sensors and gene chips are particularly relevant for directly applying the information gathered from the genome projects. In this work electrochemical techniques are used to develop methodologies to detect DNA polymorphisms in human genes using cytochrome P450 3A4 (CYP3A4) as a model gene. CYP3A4*1B oligonucleotides were immobilized on the surface of a gold electrode and hybridized with fully complementary oligonucleotide sequences as well as with mismatched sequences corresponding to the CYP3A4*1A reference sequence. The methodology developed is based on double stranded DNA's ability to transport charge along nucleotide stacking. The perturbation of the double helix pi-stack introduced by a mismatched nucleotide reduces electron flow and can be detected by measuring the attenuation of the charge transfer. The methodology developed could identify CYP3A4*1A homozygotes by the 5 microC charge attenuation observed when compared with DNA samples containing at least one CYP3A4*1B allele. PMID- 12747590 TI - Comparison of standard PCR and the LightCycler technique to determine the thrombophilic mutations: an efficiency and cost study. AB - For several years it has been possible to routinely detect numerous mutations in the human genome by different methods. The most common technique is a standard PCR, but real time fluorescence PCR is increasingly being used. The purpose of this paper is to compare these two different techniques from the point of view of reliability, time consumption, and cost. More than 600 DNA samples of prevalence studies and from cancer patients were used to determine mutations in the genes of coagulation factor V Leiden, prothrombin, and methylenetetrahydrofolate reductase using standard PCR. A subset of 132 samples from the same pool was also tested by LightCycler PCR for the same coagulation gene mutations. Originally LightCycler techniques were applied for quantitative PCR by real time fluorescence measuring. Adding a melting curve analysis allows mutation detection. The results were perfectly concordant. The cost for the reagents is nearly the same for both methods but the time consumption for standard PCR is much higher than for the LightCycler method, resulting in higher laboratory personnel costs. PMID- 12747591 TI - Impact of purified water quality on molecular biology experiments. AB - Purified water is a reagent used in a variety of molecular biology experiments, for sample and media preparation, in mobile phases of liquid chromatography techniques, and in rinsing steps. The combination of several technologies in water purification systems allows delivering high-purity water adapted to each application and technique. Through a series of examples, the importance of water quality on biotechnology experiments, such as single nucleotide polymorphism (SNP) analysis by denaturating HPLC, RNA preparation and PCR, is presented. Results obtained on DNA mutation and single nucleotide polymorphism analysis using the denaturating HPLC (DHPLC) technique highlight the benefits of organic removal by UV photooxidation process. Comparative gel electrophoresis data show that ultrafiltration is as efficient as diethylpyrocarbonate (DEPC) treatment for suppressing RNase activity in water. Gel electrophoresis and densitometry measurement also point out the benefits of ultrafiltration to carry out reverse transcriptase-polymerase chain reaction quantitatively. PMID- 12747592 TI - Which are the best tools for specific clinical application (chips, multiplex, mass spec profile, etc.)? PMID- 12747593 TI - Genetic variations observed in arterial and venous thromboembolism--relevance for therapy, risk prevention and prognosis. AB - We undertook genetic and biochemical assays in patients with arterial (n = 146) and venous (n = 199) thromboembolism and survivors of pulmonary embolism (n = 58) to study causation and gene-life style interactions. In the clinical material from North Western Russia, factor V Leiden was found to be a risk factor in venous thrombosis (OR = 3.6), while the methylenetetrahydrofolate reductase (MTHFR) C677T mutation was a significant variable in both venous (p = 0.03) and arterial thrombosis (p = 0.004). Homocysteine levels were determined (n = 84) and hyperhomocysteinemia correlated with the T allele of the MTHFR gene, and with smoking and coffee consumption. Vitamin supplementation reduced homocysteine levels dependent on MTHFR genotype (36% TT, 25% CT, 22% CC). In pulmonary embolism patients, frequency of the -455G/A beta-fibrinogen dimorphism was studied. Carriers of this allele were significantly underrepresented (p < 0.02) among pulmonary embolism survivors (34.5%) compared to controls (56.7%). Additionally, -455AA homozygotes were found in 11.7% controls but only 1.7% of pulmonary embolism patients (p = 0.006). In venous and arterial thrombosis cases, MTHFR and homocysteine data led to effective dietary supplementation with a reduced risk of disease progression. Results from the pulmonary embolism study may indicate that screening tests for the -455G/A beta-fibrinogen genetic variation could be of prognostic value, and may point the way for novel anticoagulation strategies. PMID- 12747594 TI - The TNF-alpha gene NcoI polymorphism at position -308 of the promoter influences insulin resistance, and increases serum triglycerides after postprandial lipaemia in familiar obesity. AB - Tumour necrosis factor alpha (TNF-alpha), acting as a modulator of gene expression in adipocytes, has been linked to the development of insulin resistance and obesity. The aim of this study was to investigate whether the A/G variation at position -308 in the TNF-alpha promoter influences the body weight, insulin resistance, and postprandial lipaemia in Polish Caucasians. One hundred twenty one subjects, 38 men and 83 women, representing 40 obese families, were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). TNF-1 (GG) and TNF-2 (GA and AA) allele carriers were compared with respect to body mass index, fat/lean body mass composition, waist-to-hip ratio, as well as fasting lipids, glucose, leptin, and insulin fasting, and during the oral glucose tolerance test (4 points within 2 hours) and oral lipid tolerance test (OLTT; 5 points within 8 hours). The insulin sensitivity indices HOMA-IR (homeostasis model assessment of insulin resistance), ISI-COMP (whole body insulin sensitivity index), ISI-HOMA (hepatic insulin sensitivity), and DELTA (early secretory response to an oral glucose load) were calculated. We detected 64 GG, 56 GA, and 1 AA genotypes. Significant increases of insulin resistance parameters in obese female TNF-2 allele carriers were observed (significantly increased HOMA-IR and decreased ISI-HOMA, ISI-composite). The male TNF-2 carriers were characterised by significantly increased levels of triglyceride and free fatty acids during OLTT as well as fasting glucose. The A/G variation at position -308 in the promoter region of the TNF-alpha gene could be an important genetic factor predisposing to insulin resistance in obese women and increased levels of glucose, triglyceride, and free fatty acids in men. PMID- 12747595 TI - The -308 G/A tumor necrosis factor-alpha gene dimorphism: a risk factor for unstable angina. AB - Since the inflammatory cytokine tumor necrosis factor-alpha (TNF-alpha) may play a major role in the pathophysiology of acute coronary syndromes, 299 consecutive male patients hospitalized for coronary artery disease (i.e., lumen lost > or = 50%) were genotyped for the functional -308G/A TNF-alpha polymorphism using restriction fragment length polymorphism method, in order to evaluate its potential association with the risk of unstable angina and/or myocardial infarction. A higher frequency of carriers of the A allele was observed in patients with unstable angina (n = 58) when compared to control patients with stable angina (n = 95) (39.66% vs. 23.16% respectively, p = 0.029, odds ratio = 2.2) but not in patients with myocardial infarction (n = 146) (23.97% vs. 23.16%, p = NS). Furthermore, we evidenced an interaction of the polymorphism studied with body mass index in patients with unstable angina. Thus, when stratified analysis was performed, results in patients with a body mass index < or = 27 showed a more striking association between A allele carriage frequency and unstable angina (p = 0.012, odds ratio = 3.0). These results suggest the crucial role of TNF-alpha in the mechanisms responsible for unstable angina in accordance with the concept of vulnerable plaque. On the other hand, mechanisms controlling myocardial infarction appear more complex and heterogeneous. PMID- 12747596 TI - The apolipoprotein AV gene and diurnal triglyceridaemia in normolipidaemic subjects. AB - The newly recognised apolipoprotein (apo) AV gene (APOAV) has been linked to fasting plasma triglyceride (TG) concentrations with some polymorphisms associated with elevated fasting TGs. Since fasting plasma TGs are mainly determined by the hepatic production of TG-rich particles (very low density lipoprotein; VLDL), and fasting TGs are the major determinants of postprandial lipaemia, we have evaluated the effects of an APOAV polymorphism on postprandial triglyceridaemia, which is largely determined by the intestinal production and clearance of chylomicrons. For this purpose, diurnal capillary triglyceridaemia (reflecting postprandial lipaemia) was determined in a cohort of 88 healthy volunteers (48 males and 40 females) in relation with a -1131T>C variant in the promoter of APOAV. Thirteen of these subjects (7 males and 6 females) were carriers of the -1131C allele, which has been associated with higher fasting plasma TG levels. The carriers had higher fasting capillary TG concentrations, although plasma TGs were not significantly different from non-carriers in this cohort. Surprisingly, total diurnal triglyceridaemia calculated as the area under the capillary TG curve was similar in carriers compared to non-carriers but after correction for fasting capillary TG levels, incremental diurnal triglyceridaemia was significantly lower in carriers (1.74 (5.27) mmol/h/l) than in non-carriers (4.91 (4.90) mmol/h/l; p = 0.036). The same trends were found for both males and females when analysed separately. Since dietary intake, which is a major determinant of incremental diurnal triglyceridaemia, did not differ between the two groups, we believe that these differences are at least partly explained by the APOAV. In summary, the APOAV assessed by means of the -1131T>C variant seemed to have a paradoxical effect on postprandial lipaemia when compared to fasting TG levels. PMID- 12747597 TI - Left ventricular size, mass and function in relation to angiotensin-converting enzyme gene and angiotensin-II type 1 receptor gene polymorphisms in patients with coronary artery disease. AB - The objective of the present study was to analyse the potential synergistic influence of the insertion/deletion polymorphism of the angiotensin-converting enzyme gene (I/D ACE) and the A1166C polymorphism of the angiotensin-II type 1 receptor gene polymorphisms (A1166C AT1R) on the left ventricular size and performance. Three hundred sixty and one consecutive, Caucasian patients with angiographically confirmed coronary artery disease (CAD) were enrolled into the study. Left ventricular diameter, mass and function were evaluated by echocardiography. Screening for the I/D ACE and A1166C AT1R genotypes was performed by polymerase chain reaction of genomic DNA, followed by restriction enzyme digestion and agarose gel electrophoresis. The I/D ACE and A1166C AT1R genotypes separately were not significantly associated with the left ventricular size and function parameters in CAD patients. However, trends towards decreased left ventricular ejection fraction (LVEF) as well as increased left ventricular end-diastolic diameter (LVEDD) and left ventricular mass index (LVMI) were observed when patients with genotype DD+CC/AC and DD+CC were compared to patients homozygous only in one locus (DD or CC). Significant increase in LVEDD and LVMI was observed only in patients with a history of anterior myocardial infarction with combined genotype DD+CC/AC or DD+CC. This study does not support the role of the ACE I/D and AT1R A1166C polymorphisms in the determination of the left ventricular size and performance in patients with significant coronary atherosclerosis. However, it indicates that the influence of polymorphisms may be present in specific patient populations. PMID- 12747599 TI - PON1-192 phenotype and genotype assessments in 918 subjects of the Stanislas cohort study. AB - This study describes the factors of variation of the enzymes related to the PON1 192 phenotype assessment, i.e., basal paraoxonase, salt-stimulated paraoxonase and arylesterase activities, and compares the PON1-192 phenotype to the PON1-192 genotype assessments in supposedly healthy subjects issued from the Stanislas cohort study. The studied population included 918 subjects, i.e., 221 families including 441 adults and 477 children aged 4 to 58 years. Potential determinants such as age, gender, body mass index, alcohol and tobacco consumption, and oral contraceptive intake have been studied. The PON ratio (salt-stimulated paraoxonase/arylesterase) was trimodally distributed and the cut-off values used to differentiate the two homozygous (AA and BB phenotypes) from the heterozygous (AB phenotype) subjects were 3.0 and 7.0 in this study. In males, basal paraoxonase and salt-stimulated paraoxonase activities were not affected by alcohol consumption and current smoking, but basal paraoxonase activity was decreased by 15% by current smoking and was increased by 15% by oral contraceptive intake in females as was the salt-stimulated paraoxonase activity. The level of discordance between phenotype and genotype assessments was 7.2% (66/918). Most of the discrepancies were observed between the BB and AB phenotypes (4.25%). PMID- 12747598 TI - Analysis of multiple single nucleotide polymorphisms of candidate genes related to coronary heart disease susceptibility by using support vector machines. AB - Coronary heart disease (CHD) is a complex genetic disease involving gene environment interaction. Many association studies between single nucleotide polymorphisms (SNPs) of candidate genes and CHD have been reported. We have applied a new method to analyze such relationships using support vector machines (SVMs), which is one of the methods for artificial neuronal network. We assumed that common haplotype implicit in genotypes will differ between cases and controls, and that this will allow SVM-derived patterns to be classifiable according to subject genotypes. Fourteen SNPs of ten candidate genes in 86 CHD patients and 119 controls were investigated. Genotypes were transformed to a numerical vector by giving scores based on difference between the genotypes of each subject and the reference genotypes, which represent the healthy normal population. Overall classification accuracy by SVMs was 64.4% with a receiver operating characteristic (ROC) area of 0.639. By conventional analysis using the chi2 test, the association between CHD and the SNP of the scavenger receptor B1 gene was most significant in terms of allele frequencies in cases vs. controls (p = 0.0001). In conclusion, we suggest that the application of SVMs for association studies of SNPs in candidate genes shows considerable promise and that further work could be usefully performed upon the estimation of CHD susceptibility in individuals of high risk. PMID- 12747600 TI - Lipoprotein lipase gene polymorphisms in Croatian patients with coronary artery disease. AB - Modifications in lipoprotein lipase levels lead to elevated triglycerides and reduced high density lipoprotein (HDL), both of which are risk factors for coronary artery disease (CAD). Hence, we examined the influence of the -93T/G, D9N, N291S, and S447X polymorphisms in the lipoprotein lipase (LPL) gene on CAD risk and lipid levels in Croatian patients with and without angiographically confirmed CAD. The N291S polymorphism was significantly associated with CAD (OR = 0.36; 95% CI = 0.13, 0.99; p = 0.048). This association was only moderately affected by adjusting for various lipids (OR = 0.36; 95% CI = 0.12, 1.08; p = 0.068). HDL2-cholesterol and apolipoprotein A-I levels were significantly higher in non-carriers of the -93T/G and D9N polymorphisms in the CAD group (p = 0.017 and 0.028, respectively). The N291S genetic variant did not show any significant difference between carriers and non-carriers in either group studied for any of the lipids. Lower triglyceride and higher HDL2-cholesterol levels in the control group were associated with carriers of the S447X mutation (p = 0.043 and 0.056, respectively). LPL gene polymorphisms might be involved in predisposition to CAD and determination of lipid profiles. PMID- 12747601 TI - Homocysteine, methylenetetrahydrofolate reductase C677T polymorphism and the B vitamins: a facet of nature-nurture interplay. AB - BACKGROUND: Methylenetetrahydrofolate reductase 677 (MTHFR 677) polymorphism may provoke hyperhomocysteinemia when folate status is low. The influence of MTHFR 677 mutation on homocysteine (HCY) levels in relation to vitamin B12 and folate status was investigated in the current study. SUBJECTS AND METHODS: 113 vegetarians, 123 omnivorous Germans, and 117 omnivorous Syrians were recruited. MTHFR 677 genotype, HCY, methylmalonic acid (MMA), total serum vitamin B12, serum folate, and vitamin B6 were determined using conventional methods. RESULTS: Omnivorous Germans displayed the lowest HCY levels compared with vegetarians and Syrians (median 8.0, 10.4, and 11.3 micromol/l, respectively). The highest serum folate and the highest MMA levels were found in vegetarians (median folate = 30.0; MMA = 355 nmol/l). Among vegetarians and Syrians, TT subjects had higher HCY levels than other genotypes which were, however, no longer significant in the highest folate tertiles. When the data were pooled, the odds ratio (OR, 95% CI) for HCY > 12 micromol/l was 3.81 (1.55-9.34) in TT compared with CC subjects. The OR increased to 28.85 (4.63-179.62) in TT subjects who had folate in the lowest tertile, and to 21.84 (4.81-99.1) in TT subjects who had MMA in the highest MMA tertile. CONCLUSION: MTHFR 677 TT individuals are more liable to hyperhomocysteinemia under vitamin B12 deficiency than the other two genotypes. In such a case, relative folate shortage may progressively increase HCY levels. TT individuals may have increased folate and vitamin B12 requirements compared to the other CC and CT genotypes. PMID- 12747602 TI - Which, and how limited number of polymorphisms should be selected per disease, risk assessment, health profile or biological system? PMID- 12747603 TI - Ethical issues: should we give the predictive genetic profile to the citizens? PMID- 12747604 TI - Pharmacogenetics and responders to a therapy: theoretical background and practical problems. AB - In a narrow meaning, responders to a therapy are all those who will react as expected following the administration of this therapy. However, a wider definition is worth considering: all those for whom the administration of the therapy will be beneficial. Innovative therapies are increasingly expensive and hazardous, and limiting prescriptions to responders is both economically and ethically compulsory. The theoretical basis for such an approach exists. The process of defining the profile of responders consists of identifying the characteristics of the patients that interact with the size of the effect and integrating them quantitatively in a predictive model. The effect model, which is the relation between the risks of the event with and without the treatment, can be used for the prediction. It can integrate interactions of the efficacy with risk factors and/or genes. The data to be used to achieve both the identification of the interactions and the building of the predictive model are those from the studied population, the set of patients enrolled in clinical trials. Hence, the process of defining the therapy is an extrapolation from the studied population. To carry out the extrapolation process one can use various available techniques, of which none fully fits the purpose. No method is currently both fully adequate and validated. Finally, the predictive models, which we need to identify responders, do not exist in practice. Fortunately, new research approaches have been developed recently. PMID- 12747605 TI - Pharmacogenetics of drug metabolising enzymes: importance for personalised medicine. AB - The number of polymorphisms identified in genes encoding drug metabolising enzymes, drug transporters, and receptors is rapidly increasing. In many cases, these genetic factors have a major impact on the pharmacokinetics and pharmacodynamics of a particular drug and thereby influence the sensitivity to such drug in an individual patient with a certain genotype. The highest impact is seen for drugs with a narrow therapeutic index, with important examples emerging from treatment with antidepressants, oral anticoagulants, and cytostatics, which are metabolised by the polymorphic enzymes cytochrome P450 2D6 (CYP2D6), cytochrome P450 2C9 (CYP2C9), and thiopurine-S-methyltransferase (TPMT), respectively. In order to apply the increasing amount of pharmacogenetic knowledge to clinical practise, specific dosage recommendations based on genotypes will have to be developed to guide the clinician, and these recommendations will have to be evaluated in prospective clinical studies. Such development will lead to a patient-tailored drug therapy which hopefully would be more efficient and will result in fewer adverse drug reactions. PMID- 12747606 TI - Pharmacogenomics and pharmacogenetics of cholesterol-lowering therapy. AB - Cholesterol-lowering therapy is the central approach in the primary and secondary prevention of cardiovascular disease, the leading cause of death in industrialized countries. 3-Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) are currently the most potent and widely used cholesterol-lowering drugs. Large-scale clinical trials unequivocally demonstrated the efficacy of statin treatment in reducing the risk of cardiovascular events. In general, HMG-CoA reductase inhibitors are well tolerated, although in a minority of patients severe adverse effects like myopathy or rhabdomyolysis may develop. The incidence of this potentially life threatening side effects increases with co-adminstration of drugs that are metabolized via the same pharmacokinetic pathways or at high-dose statin therapy. The recent focus on the pleiotropic effects of statins that are more frequently observed at higher doses and the conclusion drawn from the large statin trials that low-density lipoprotein (LDL)-cholesterol is "the lower the better", may need careful consideration in individuals at risk of adverse drug reactions. On the other hand, not all patients respond to statin therapy with a reduction in coronary heart disease (CHD) risk. It is therefore of interest to develop diagnostic test systems, which would allow to identify patients at increased risk of adverse drug reactions or patients with a lack of therapeutic effect. Beside exogenous factors, genetic variability determines the response of an individual to drug therapy and the analysis of genetic variants affecting pharmacokinetic or pharmacodynamic aspects of drug therapy is the subject of pharmacogenomics. This review summarizes current knowledge of the pharmacology and the pharmacogenomics of statin therapy. PMID- 12747607 TI - Pharmacogenomics of drugs affecting the cardiovascular system. AB - The variability in drug response originates partly from genetics, with possible consequences for drug efficacy, adverse effects, and toxicity. Until now, pharmacogenetics mainly indicated the best known source of variability, that is, the variability caused by drug metabolism. However, simultaneous progress in the knowledge of biochemical targets of drugs and of the human genome, together with the development of new technologies, revealed many new sources of human genetic variation, e.g., in receptors or transporters. Drugs are metabolized by various polymorphic phase I enzymes, including cytochromes P450 (CYP). Among them, the most relevant for the metabolism of cardiovascular drugs are CYP3A4, CYP2C9 or CYP2C19, and CYP2D6. The role of phase II enzymes is limited with regard to cardiovascular drugs biotransformation, but some polymorphisms (glutathion-S transferase; GSH-T) are linked to cardiovascular risk. Phase III proteins or transporters, especially from the ABC family, must also be considered, as their polymorphisms affect cholesterol and other sterols transport. Among pharmacological targets, some proteins were identified as involved in interindividual variations in the response to cardiovascular drugs. Some examples are apolipoprotein E, angiotensin-converting enzyme, and the beta-adrenergic receptor. From the risk concept emphasizing impaired metabolism and adverse effects, we now moved to an approach, which is a personalized, genotype-dependent adaptation of therapy. PMID- 12747608 TI - Allele frequencies for glutathione S-transferase and N-acetyltransferase 2 differ in African population groups and may be associated with oesophageal cancer or tuberculosis incidence. AB - Glutathione S-transferase (GST) and arylamine N-acetyltransferase 2 (NAT2) metabolise many environmental and chemotherapeutic agents, which influence susceptibility to disease. Polymorphisms in these enzymes result in different host phenotypes and contribute to different disease profiles or responses to toxic or chemotherapeutic agents, depending on their frequency in different populations. GST and NAT2 polymorphisms were investigated in different population groups, including African populations, and a range of allelic frequencies have been observed. The GSTM1 null genotype frequency, reported in this paper in two South African ethnic groups, is the lowest reported (0.19-0.21). In contrast, these same groups have a high GSTT1 null frequency (0.41-0.54), which is considerably higher than in African-Americans, or other Africans. The GSTT1 null frequency is comparable to the Chinese, a population with a very high oesophageal cancer incidence, similar to that in the African group. The frequency of the GSTPi Val105 variant in the South African Xhosas was also high (0.53), differing significantly from the low frequency in other Africans. These variants could therefore be associated with high cancer susceptibility. In addition, the high proportion of NAT2 "fast" alleles may partially explain the high tuberculosis prevalence in South Africans, due to reduced isoniazid efficacy in the presence of rapid acetylation. PMID- 12747610 TI - Pharmacogenetics: from bench to bedside. PMID- 12747609 TI - CYP3A4*1B and NAT2*14 alleles in a native African population. AB - Single nucleotide polymorphisms were examined in the cytochrome 450 3A4 (CYP3A4) and N-acetyltransferase 2 (NAT2) genes, which code for major mediators of the metabolism of a wide variety of therapeutic drugs, as well as xenobiotics. We determined, in a population from Guinea-Bissau, the frequencies of CYP3A4 and NAT2 variants expected to be prevalent among Africans, due to the high frequency previously observed in African Americans. The observed frequencies were 72% for CYP3A4*1B and 19.2% for the NAT2 191 G>A variant. The high frequency found for these potentially function-altering polymorphisms suggests the possibility of impaired metabolism through CYP3A4 and NAT2 in this population. Strikingly, the frequency observed for the NAT2 191 G>A single nucleotide polymorphism (SNP), associated with the slow acetylator phenotype, was significantly higher than found in other African populations, suggesting the existence of a west to east gradient across Sub-Saharan Africa. The prevalence of these variants may be relevant with regard to therapeutic efficacy in African populations for it may potentially affect drug clearance and consequently, increase the incidence of side effects and drug-drug interactions. PMID- 12747611 TI - Investigation of radiosurgical beam profiles using Monte Carlo method. AB - An accurate determination of the penumbra of radiosurgery profiles is critical to avoid complications in organs at risk adjacent to the tumor. Conventional detectors may not be accurate enough for small field sizes. The Monte Carlo (MC) method was used to study the behavior of radiosurgical beam profiles at the penumbral region; the BEAM code was also used in this work. Two collimators (2.2- and 0.3-cm diameter) were calculated and compared with empirical measurements obtained with the detectors normally used. The differences found between film dosimetry and MC revealed a systematic error in the reading procedure. In the process, a water phantom was simulated with a layer of the same composition as that of the film. MC calculations with film differed by a small amount from those obtained with the water phantom alone. In conclusion, MC may be used as a verification tool to support dosimetrical procedures with conventional detectors, especially in very small beams such as those used in radiosurgery. Furthermore, it has been proved that the film energy dependence is negligible for fields used in radiosurgery. PMID- 12747612 TI - Patient training in respiratory-gated radiotherapy. AB - Respiratory gating is used to counter the effects of organ motion during radiotherapy for chest tumors. The effects of variations in patient breathing patterns during a single treatment and from day to day are unknown. We evaluated the feasibility of using patient training tools and their effect on the breathing cycle regularity and reproducibility during respiratory-gated radiotherapy. To monitor respiratory patterns, we used a component of a commercially available respiratory-gated radiotherapy system (Real Time Position Management (RPM) System, Varian Oncology Systems, Palo Alto, CA 94304). This passive marker video tracking system consists of reflective markers placed on the patient's chest or abdomen, which are detected by a wall-mounted video camera. Software installed on a PC interfaced to this camera detects the marker motion digitally and records it. The marker position as a function of time serves as the motion signal that may be used to trigger imaging or treatment. The training tools used were audio prompting and visual feedback, with free breathing as a control. The audio prompting method used instructions to "breathe in" or "breathe out" at periodic intervals deduced from patients' own breathing patterns. In the visual feedback method, patients were shown a real-time trace of their abdominal wall motion due to breathing. Using this, they were asked to maintain a constant amplitude of motion. Motion traces of the abdominal wall were recorded for each patient for various maneuvers. Free breathing showed a variable amplitude and frequency. Audio prompting resulted in a reproducible frequency; however, the variability and the magnitude of amplitude increased. Visual feedback gave a better control over the amplitude but showed minor variations in frequency. We concluded that training improves the reproducibility of amplitude and frequency of patient breathing cycles. This may increase the accuracy of respiratory-gated radiation therapy. PMID- 12747613 TI - On the practice of the clinical implementation of enhanced dynamic wedges. AB - Practical aspects of the clinical implementation of enhanced dynamic wedges (EDW) replacing manual wedges are presented and discussed extensively. A comparison between measured and calculated data is also presented. Relative dose distributions and wedge factors were calculated with a commercially available treatment planning system and measured in a water-phantom and with an ionization chamber. Wedge factor calculations and measurements were also compared with an independent method of wedge factor calculations available from the literature. Aspects of the clinical implementation, such as safety and quality assurance, were evaluated. Measurements and calculations agreed very well and were slightly better than results of previous studies. Profiles and percentage depth doses (PDDs) agreed within 1% to 1.5% and within 0.5%, respectively. Measured and calculated wedge factors ratios agreed within 0.5% to 1%. Calculated and measured EDW dose distributions showed excellent agreement, both relative and absolute. However, for safe and practical use, specific aspects need to be taken into consideration. Once the treatment planning system is commissioned properly, the clinical implementation of EDW is rather straightforward. PMID- 12747614 TI - A comparison of methods to calculate biological effectiveness (RBE) from Monte Carlo simulations. AB - The relative biological effectiveness (RBE) of radiation is assessed and easily calculated by Monte Carlo simulations of the passage of radiation through matter. The expression to calculate the RBE provided by microdosimetry requires the use of the energy spectrum of charged particles. This paper compares the RBE values obtained for Palladium-103 (103Pd) and iodine-125 (125I) when calculated with 2 different spectra: the electron slowing-down spectrum and the ejection spectrum. The former yields a value of 10.6%, twice the value obtained with the latter (4.5%). Which spectrum to use is an open question. A theoretical argument is presented in favor of the ejection spectrum. PMID- 12747615 TI - Effect of bite tray impression technique on relocation accuracy in frameless stereotactic radiotherapy. AB - A previously developed method for achieving patient relocation in fractionated stereotactic radiotherapy (attachment of an infrared fiducial system to a bite tray) relies on the integrity of a bite tray system that incorporates moulding to the patient's upper dentition. Reproducible and accurate patient positioning requires stability of the bite tray and mould during the full treatment process, both during the time the bite tray is inserted in the patient's mouth, and between separate bite tray insertions. The optimum construction method for a stable reproducible tray has not been sufficiently investigated. We undertook a study to identify factors which might influence the integrity of the hard palate bite tray system. Reprosil Fast Set Putty was used to construct 3 impression conditions; teeth only; teeth and alveolar sulcus; and teeth, alveolar sulcus, and the hard palate. Reproducibility was assessed by volunteers inserting the impressions multiple times and recording the locations of 8 standard reference points. Our results showed the optimal impression technique (i.e., the one that led to the smallest ranges in positional and rotational errors) was that which incorporated the teeth, alveolar sulcus, and hard palate. PMID- 12747616 TI - Total skin electron irradiation: evaluation of dose uniformity throughout the skin surface. AB - In this study, in vivo dosimetic data of 67 total skin electron irradiation (TSEI) treatments were analyzed. Thermoluminescent dosimetry (TLD) measurements were made at 10 different body points for every patient. The results demonstrated that the dose inhomogeneity throughout the skin surface is around 15%. The homogeneity was better at the trunk than at the extratrunk points, and was worse when a degrader was used. There was minimal improvement of homogeneity in subsequent days of treatment. PMID- 12747617 TI - Craniospinal treatment with the patient supine. AB - Radiotherapy of the craniospinal axis in young children is frequently complicated by the need for access to the patient's airway for sedation and anesthesia delivery or by frequent, unanticipated movement. Positioning the patient supine, instead of in the conventional prone position, allows the use of immobilization facemasks with body molds and more positive patient fixation, and improved airway access. The procedure for establishing the various fields differs from the prone approach. In this paper, we describe the methodology to achieve successful supine positioning. PMID- 12747618 TI - Is one head and neck immobilization system as good as another? One center's experience. AB - The William Buckland Radiotherapy Center has used 2 different immobilization systems for patients requiring radiotherapy to the head-and-neck region. A polycarbonate mask was manufactured for radical treatments and a thermoplastic mask for palliative treatments. This study evaluated field placement accuracy, staff opinion, and production costs of both systems. The manual matching program of Varian PortalVision Electronic Portal Imaging (EPI) System was used to assess field placement accuracy on a daily basis. Radiation therapists (RTs) were surveyed before and after the study to determine their opinions of each system. Production time and required materials were recorded to assess cost. Nineteen patients from each system had daily EPI results compiled with no statistically significant difference observed in field placement accuracy. The thermoplastic system was found to be more cost efficient due to a combination of the reduced production time and reuseability of the masks. User acceptability of the thermoplastic system has increased so that it is now the preferred system. In conclusion, the thermoplastic system is a viable alternative to the polycarbonate system in terms of treatment accuracy and cost. It is recommended that the thermoplastic system be used for all radical and palliative treatments. In addition, RTs prefer the thermoplastic system. PMID- 12747619 TI - Feasible measurement errors when undertaking in vivo dosimetry during external beam radiotherapy of the breast. AB - In vivo dosimetry is a proven reliable method of checking overall treatment accuracy, allowing verification of dosimetry and dose calculation as well as patient treatment setup. We conducted a pilot study to assess the clinical utility of in vivo dosimetry in our department. Diodes (calibrated for typical treatment conditions) were used to record entrance dose measurements on 62 patients representing a variety of treatment sites. Measurements were compared with predictions from the planning system, with results found to be in tolerance for the majority of treatment sites. However, large discrepancies were encountered for measurements performed during breast irradiation (up to 16% for lateral tangential fields). The sensitivity of the recorded entrance dose to the positioning error of the diode placement was examined. The sensitivity of diode signal to small changes in position were compared with feasible variations in other parameters (e.g., dosimetry, FSD at setup). For the breast irradiation technique considered, wedges are used for the majority of fields. It was found that a proportion of error was predominantly due to the use of wedges and the presence of significantly nonuniform patient contours. In combination with diode placement errors, this resulted in increased measurement error. Correct diode placement is critical to ensure accurate data collection. The results of this study indicate the importance of separating errors due to measurement technique from actual treatment/setup errors. PMID- 12747620 TI - Evaluation of the veridose QC phantom. AB - Daily, monthly, and annual quality control (QC) of linear accelerators are part of the major tasks of the medical physicist to verify that patients are receiving proper radiation treatment. The control tests consist of the measurement of beam output, verification of the beam energy, and determination of the beam flatness and symmetry in a linear accelerator. A new device, referred to as QC phantom, was designed and fabricated for the QC of linear accelerators. This device is accompanied by software generating the reports of all measured data, keeping track of day-to-day data, and plotting the results. The accuracy, reproducibility, and linearity of the QC phantom were evaluated in this project. Also, the user friendliness of this device for morning warmup of linear accelerators was tested. PMID- 12747621 TI - Bisphosphonate mucosal injury--the end of the story? PMID- 12747622 TI - Helicobacter pylori infection: the diagnostic dilemma is still going on! PMID- 12747623 TI - Endoscopic 13C-urea breath test for the diagnosis of Helicobacter pylori infection. AB - BACKGROUND: Endoscopic 13C-urea breath test may avoid contamination of oral urease and rapidly discriminate Helicobacter pylori-positive and Helicobacter pylori-negative patients. AIMS: To compare the accuracy of endoscopic 13C-urea breath test with conventional invasive methods in diagnosis of Helicobacter pylori infection. PATIENTS: One hundred patients who attended for routine upper gastrointestinal endoscopy were included. METHODS: 13C-urea was applied to the stomach through the working channel of endoscope at the end of endoscopic examination. Breath samples were collected before endoscopy and 2, 4, 6, 8, 10 min after consumption of 100 or 50 mg 13C-urea. Helicobacter pylori infection was defined as those with positive culture or positive results of both histology and CLO test. RESULTS: The accuracy of 100 mg endoscopic 13C-urea breath test was significantly higher than that of culture and CLO test (100% vs. 88% and 92%, p = 0.02 and 0.03, respectively). The accuracy of 50 mg endoscopic 13C-urea breath test was higher than that of histology and CLO test (98% vs. 90% and 96%, respectively), although the differences were not statistically significant. CONCLUSIONS: Endoscopic 13C-urea breath test has a higher accuracy compared with biopsy-based modalities. It may be a good choice to diagnose Helicobacter pylori infection if endoscopy is indicated for a dyspeptic patient. PMID- 12747624 TI - Change in apoptosis in the gastric surface epithelium and glands after eradication of Helicobacter pylori. AB - BACKGROUND: Change in apoptosis in gastric glands after eradication of Helicobacter pylori has never been reported. AIMS: The purpose of this paper is to investigate the change in apoptosis in gastric glands after eradication of Heliobacter pylori. PATIENTS AND METHODS: We studied 23 Heliobacter pylori positive patients with duodenal and gastric ulcers, who were monitored for 6-12 months after eradication, and eight controls. Biopsies were taken from the antrum and body. Apoptosis was evaluated immunohistochemically using anti-single stranded DNA antibody. Apoptotic index was calculated by counting immunostained cells in surface epithelial and glandular cells. RESULTS: In the surface epithelium, Apoptotic indexes were significantly higher in patients than in controls. In the upper portion of fundic glands, apoptotic indexes were significantly higher in patients with gastric ulcers (14.2% (9.3, 17.8)) (median (1st quartile, 3rd quartile)) than in controls (8.0% (2.0, 9.0), p < 0.01) and decreased significantly after eradication (3.4% (2.0, 5.3)), p < 0.01). In pyloric glands, apoptotic indexes were no different between patients and controls. In the lower portion of fundic glands, apoptotic indexes were very low, both in patients and in controls. CONCLUSIONS: Our results showed that apoptosis, not only of surface epithelial cells but also of glandular cells in the upper portion of fundic glands, increased in Heliobacter pylori-positive patients with gastric ulcers and decreased to normal levels after eradication of Heliobacter pylori. PMID- 12747625 TI - Oral contraceptives and colorectal cancer. AB - BACKGROUND: Only limited information and understanding are available on the potential relation between oral contraceptive use and the risk of colorectal cancer. Further data on the issue are therefore useful and may help informed choice of contraception. METHODS AND RESULTS: Data were derived from a case control study of colorectal cancer conducted between 1992 and 2001 in the Swiss Canton of Vaud, including 131 women with colorectal cancer and 373 controls admitted in the same hospital as the cases with diagnosis of acute, non neoplastic disease, unrelated to long-term modification of diet. Oral contraceptive use was reported by 11% of cases versus 17% of controls, corresponding to multivariate odds ratio of 0.8 (95% CI: 0.4-1.7). The odds ratio was non-significantly below unity across strata of duration, time since first and last oral contraceptive use. CONCLUSION: These findings add further evidence on a possible inverse relation between oral contraceptive and colorectal cancer risk. PMID- 12747626 TI - Basic study of an agent for reinforcement of near-infrared fluorescence on tumor tissue. AB - BACKGROUND AND AIMS: An indocyanine green derivative (ICG-sulfo-OSu) and agents for reinforcement of infrared fluorescence, which can be used as an infrared fluorescent labeling substance suitable for detection of microlesions by an IR fluorescence endoscope, have been developed. The study aims were to confirm the ability of a reinforcement agent, as well as imaging processing, to intensify fluorescence from the labeled antibody on immunohistochemical staining. SUBJECTS AND METHODS: ICG-sulfo-OSu-labeled MUC1 antibody and an IR fluorescence imaging system were employed in the present study. Paraffin sections of gastric cancer were stained with anti-MUC1 antibody by the avidin-biotinylated peroxidase complex method. Among the positive specimens, three cases were used for IR imaging analysis. Octylglucoside was used as a reinforcement agent. RESULTS: The incubation of paraffin sections with ICG-sulfo-OSu-labeled MUC1 antibody resulted in positive staining of the tumor sites by an IR fluorescence imaging system, and the intensity of fluorescence was increased depending on the concentration of octylglucoside and grade of imaging processing. CONCLUSION: A reinforcement agent, and image processing, intensify a labeled antibody excitable by infrared fluorescence in tumor sections and can generate a strong enough fluorescent signal to detect small cancers when examined with an infrared fluorescence endoscope. PMID- 12747627 TI - Gastric metaplasia and chronic inflammation at the duodenal bulb mucosa. AB - BACKGROUND: Chronic inflammation and gastric metaplasia are often observed in biopsy specimens from the duodenal bulb of Heliobacter pylori positive patients with duodenal ulcer disease (DU). AIMS: We set out to investigate the prevalence of these lesions and their associations with other gastric and duodenal histopathological lesions. PATIENTS: A total of 1255 consecutive patients who underwent upper gastrointestinal endoscopy were recruited into the present study. METHODS: Two biopsy specimens were obtained from each of the following sites: duodenal bulb, gastric antrum, gastric body, and distal to the superior duodenal angle. These specimens were stained with hematoxylin-eosin, alcian blue periodic acid Schiff (pH 2.5) and modified Giemsa (Heliobacter pylori infection was determined only by histology). RESULTS: The mean age of the study population was 57 years, and male:female ratio 1:1.6. Overall, 235 (19%) had gastric metaplasia and/or chronic inflammation in the duodenal bulb mucosa, and H. pylori organisms could be found in 17 (1%). In univariate analyses, gastric metaplasia and/or chronic duodenal bulb inflammation positively associated with male sex (p = 0.046), Heliobacter pylori-positive chronic gastritis (p = 0.033), villous atrophy of distal duodenal mucosa, i.e., coeliac disease (p < 0.001), duodenal ulcer (p < 0.001), and duodenal bulb deformity and scarring in endoscopy (p < 0.001), but not with age (p = 0.7) nor use of nonsteroidal anti-inflammatory drugs (p = 0.055). Multivariate analysis revealed that independent risk factors for gastric metaplasia and chronic inflammation in duodenal bulb were duodenal Heliobacter pylori infection (odds ratio 1.6, 95% confidence interval CI 1.1 2.1), and villous atrophy of the distal duodenal mucosa (odds ratio 12.7, 95% CI 4.4-36.5), while chronic atrophic gastritis was protective against them (odds ratio 0.5, 95% CI 0.3-0.8). CONCLUSIONS: In addition to Heliobacter pylori infection, duodenal bulb gastric metaplasia and chronic inflammation may result from predisposition to toxic dietary components in gluten-sensitive subjects. PMID- 12747628 TI - Continuing low incidence of Crohn's disease in Northwest Greece. AB - AIM OF STUDY: The largest population-based study for inflammatory bowel disease in Northwest Greece. MATERIALS AND METHODS: A retrospective survey for the years 1982-1997. RESULTS: Of 400 patients, 334 had ulcerative colitis, 43 Crohn's disease and 23 indeterminate colitis. CONCLUSIONS: Crohn's disease still remains rare in Northwest Greece. PMID- 12747629 TI - Natural course of acute hepatitis C: a long-term prospective study. AB - BACKGROUND: Acute hepatitis C has a high chronicity rate which appears to be significantly reduced by early antiviral treatment. However, it is unclear if all acutely infected patients should be treated, and when. In this prospective study, patients with a well-documented diagnosis of acute hepatitis C were evaluated to define the natural course, the rate of chronicity, and host and virus-related factors which might predict a self-limiting or chronic evolution requiring early antiviral treatment. METHODS: From 1995 to 2000, 40 consecutive patients with a community-acquired AHC were enrolled. Liver tests, anti-hepatitis C virus antibodies and hepatitis C virus RNA levels were monitored. Median follow-up was 35 months (range 12-68). RESULTS: A total of 24/40 patients had symptomatic disease including 20 with jaundice; 13/40 patients had prompt serum hepatitis C virus RNA clearance and ALT normalisation within 12 weeks; in 12/13 patients this pattern remained unchanged during follow-up. Overall, 27/40 patients remained hepatitis C virus RNA positive with fluctuating ALT levels. Older age and jaundice were predictive of resolution whereas there was no correlation with other host factors, viral genotype or viral load. CONCLUSIONS: Our data demonstrate that spontaneous resolution can occur in about 30% of AHC patients. This favourable outcome rarely occurs in patients with anicteric AHC or in those with jaundice but with persistent viremia for more than 12 weeks from onset; early antiviral treatment for these patients may avoid or reduce chronicity. PMID- 12747631 TI - Pathways of cholesterol crystallization in model bile and native bile. AB - Hypersecretion of hepatic cholesterol, chronic supersaturation of bile with cholesterol and rapid precipitation of cholesterol crystals in the gallbladder from cholesterol-enriched vesicles represent the primum movens in cholesterol gallstone formation. Physical-chemical factors and pathways leading to cholesterol crystallization can be investigated in artificial model biles and ex vivo in fresh human bile. Depending on modulatory factors (i.e., lipid concentration, bile salt or phospholipid species, humidity, mucins, etc.), cholesterol can precipitate in several forms (i.e., monohydrate, anhydrous) and habits (i.e., plate-like, needle-like, intermediate arcs, filaments, tubules, spirals). Careful analysis of biliary cholesterol crystals includes biochemical analysis of precipitated crystals, polarizing quantitative light microscopy, and turbidimetric methods. In this paper, recent concepts on cholesterol crystallization in artificial model biles as well as in human bile will be reviewed. PMID- 12747630 TI - A puzzling jaundice. AB - In an adult beta-thalassaemic patient admitted on account of the sudden onset of nausea, vomiting and biliary pain, the concomitant jaundice was considered "obstructive" on the basis of ultrasonographic evidence of cholecysto-choledocal stones. However, when the patient was re-evaluated it was found that concomitant Gilbert's disease, hepatitis C virus related cirrhosis and dilated cardiomyopathy had a detrimental effect on clinical and laboratory findings. PMID- 12747632 TI - Helicobacter pylori stool antigen test before and after eradication therapy. PMID- 12747633 TI - Primary biliary cirrhosis-autoimmune hepatitis overlap syndrome: a cause of resistance to ursodeoxycholic treatment. PMID- 12747634 TI - Controlled bioassay systems for determination of lethal infective doses of tissue homogenates containing Taura syndrome or white spot syndrome virus. AB - In vivo bioassay is the predominant method for evaluating the infectivity of materials potentially harboring viable shrimp pathogens and determining the relative susceptibility of shrimp species to viral infections. A controlled bioassay system for white spot syndrome virus (WSSV) and Taura syndrome virus (TSV) was developed utilizing 260 ml tissue culture flasks modified with an air exchange vent. Individual shrimp (1.00 +/- 0.25 g) were placed in separate flasks containing artificial seawater (100 to 150 ml) and held in an incubator at 27 degrees C. After a 48 h acclimation period, shrimp were either injected intramuscularly with viral inoculum or exposed to virus-laden water. Water was exchanged and shrimp were fed a commercial food pellet daily except 24 h post infection (p.i.). Bioassays were performed with serial dilutions of stock viral preparations and shrimp mortality was recorded for 7 d p.i. Mortality rates of test animals permitted the estimation of the lethal infective doses, LD50 and LD90. The LD50 of the TSV injection preparation was estimated at viral dilutions of 1:7.692 x 10(7) (Trial 1) and 1:6.667 x 10(7) (Trial 2). The LD50s of 2 different WSSV injection preparations were estimated at 1:4.444 x 10(6) and 1:4.505 x 10(6). The LD50 for the TSV waterborne challenge was 1:9916 (Trial 1) and 1:15 710 (Trial 2) at 20 degrees C and 1:1272 at 27 degrees C. A second waterborne TSV inoculum challenge at 27 degrees C produced an LD50 of 1:2857. WSSV doses used in the waterborne challenge only reached 39% mortality, which did not allow for the estimation of effective lethal doses. Bioassay by injection proved to be a more reliable method of estimating viral infectivity compared to waterborne method. The dose-response curves developed can serve as a basis for controlled comparisons of relative levels of viral infectivity of specific tissue preparations and for controlled comparisons of relative susceptibility of shrimp species or stocks to viral pathogens. PMID- 12747635 TI - Relationship between white spot syndrome virus and indicators of quality in Penaeus monodon postlarvae in Karnataka, India. AB - White spot disease (WSD) is a viral disease of shrimp caused by white spot syndrome virus (WSSV). Stocking WSSV-infected seed has been implicated as a major risk factor for outbreaks of WSD. In addition, the quality of postlarvae batches has been proposed as a predictor for good crops. This paper describes the relationship between indicators of quality and WSSV in postlarvae (PL) of Penaeus monodon from Karnataka, India, over the period September 1999 to January 2000. Three outcome variables were considered: the WSSV status of the PL, as determined by PCR, and 2 subjective assessments of PL quality, namely the activity of the PL and the quality of the PL as determined by research assistants and farmers, respectively. Of the 73 batches of PL, 49.3% from a random sample of farms tested positive for WSSV. After adjusting for confounding, stocking earlier in the growing season and duration of transportation were the main risk factors for the presence of WSSV. The quality assessed by farmers and the PL activity assessed by research assistants showed only fair agreement (kappa 0.252) reaffirming the subjective nature of such techniques. The only variables consistently associated with either assessment of quality in univariate analysis were PL length, number per bag and salinity of the water in the delivery bags. After adjusting for confounding, no single variable was consistently associated with PL quality and activity. The research assistants' assessment of PL activity was also associated with the hatchery and a brown-orange hepatopancreas in univariate analysis. After adjusting for confounding, a brown-orange hepatopancreas was still significant and fitted into the model together with the salinity of the water in the PL bags. The farmers' assessment of quality was associated with PL length, date of stocking and duration of transportation in both univariate and multivariable analyses. There was no relationship between quality assessment and WSSV in PCR positive PL. PMID- 12747636 TI - Experimental mycobacteriosis in striped bass Morone saxatilis. AB - Striped bass Morone saxatilis were infected intraperitoneally with approximately 10(5) Mycobacterium marinum, M. shottsii sp. nov., or M. gordonae. Infected fish were maintained in a flow-through freshwater system at 18 to 21 degrees C, and were examined histologically and bacteriologically at 2, 4, 6, 8, 17, 26, 36 and 45 wk post-infection (p.i.). M. marinum caused acute peritonitis, followed by extensive granuloma development in the mesenteries, spleen and anterior kidney. Granulomas in these tissues underwent a temporal progression of distinct morphological stages, culminating in well-circumscribed lesions surrounded by normal or healing tissue. Mycobacteria were cultured in high numbers from splenic tissue at all times p.i. Standard Ziehl-Neelsen staining, however, did not demonstrate acid-fast rods in most early inflammatory foci and granulomas. Large numbers of acid-fast rods were present in granulomas beginning at 8 wk p.i. Between 26 and 45 wk p.i., reactivation of disease was observed in some fish, with disintegration of granulomas, renewed inflammation, and elevated splenic bacterial densities approaching 10(9) colony-forming units g(-1). Infection with M. shottsii or M. gordonae did not produce severe pathology. Mild peritonitis was followed by granuloma formation in the mesenteries, but, with 1 exception, granulomas were not observed in the spleen or anterior kidney. M. shottsii and M. gordonae both established persistent infections in the spleen, but were present at densities at least 2 orders of magnitude less than M. marinum at all time points observed. Granulomas in the mesenteries of M. shottsii- and M. gordonae infected fish resolved over time, and no reactivation of disease was observed. PMID- 12747637 TI - Carriage of potentially fish-pathogenic bacteria in Sparus aurata cultured in Mediterranean fish farms. AB - A bacteriological survey of gilthead sea bream Sparus aurata from different fish farms and culture systems on the Spanish Mediterranean coast was conducted. Three different studies were performed. Study A included hatchery-reared larvae; Study B, periodic examination of randomly sampled growing fish; and Study C, growing fish sampled only during mortality/morbidity events. In Studies B and C, sea cages, earth ponds and indoor tanks were surveyed, and in both cases diseased (showing clinical signs) and non-diseased fish were included. In Study A, a shift from Vibrio spp. (30 d after hatching) to oxidative species (60 d after hatching) was detected, and no mortality events were registered. The percentage of fish yielding bacterial growth were similar in Studies B and C, reaching 57.4 and 61.3%, respectively. A statistically significant relationship between the bacterial carriage and the type of facility was only found in Study B, showing that fish from sea cages had a higher bacterial occurrence than fish from other facilities. A statistically significant relationship between bacterial carriage and signs of disease was found, although the pattern differed in each study. Thus, in Study B only 36.2% of fish yielding abundant bacterial growth were diseased, versus 68.0% in Study C. In total, 25.0% of the fish examined were diseased. Bacterial species composition was similar in asymptomatic and diseased fish, except for a group of V. ichthyoenteri-like isolates that occurred almost exclusively in asymptomatic fish. Dominant bacterial species were V. harveyi and V. splendidus, followed by V. ichthyoenteri-like isolates, Photobacterium damselae ssp. damselae and V. fisheri. Non-fermenters were less frequent but, among them, unidentified halophilic Cytophaga-Flavobacterium isolates and Pseudoalteromonas haloplanktis were the most abundant. An association of individual species with disease was not clear, which suggests the involvement of mixed infections. PMID- 12747638 TI - Development of sensitive, high-throughput one-tube RT-PCR-enzyme hybridisation assay to detect selected bacterial fish pathogens. AB - Bacterial monitoring and surveillance is critical for the early detection of pathogens to avoid the spread of disease. To facilitate this, an efficient, high performance and high-throughput method to detect the presence of femotgram amounts of ribosomal RNA from 4 bacterial fish pathogens: Aeromonas salmonicida; Tenacibaculum maritimum (formerly Flexibacter maritimus); Lactococcus garvieae; and Yersinia ruckeri was developed. The system uses NucleoLink strips for liquid- and solid-phase PCR in 1 tube, to perform RT-PCR-enzyme hybridisation assays (RT PCR-EHA) detecting 4 fg or less of rRNA from pure cultures and between 1 and 9 CFU per 200 microl sample volume from selective-enrichment culture media. The liquid-phase amplicons were visualised by gel electrophoresis and the solid-phase amplicons detected using internal probes and visualised using colorimetric detection and p-nitrophenylphosphate. PMID- 12747639 TI - Infectivity and pathogenicity of the oomycete Aphanomyces invadans in Atlantic menhaden Brevoortia tyrannus. AB - Atlantic menhaden Brevoortia tyrannus develop characteristic skin ulcers in response to infection by the oomycete Aphanomyces invadans. To investigate pathogenicity, we conducted a dose response study. Juvenile menhaden were inoculated subcutaneously with 0, 1, 5, 10, 100, and 500 secondary zoospores per fish and monitored for 37 d post-injection (p.i.). Survival rates declined with increasing zoospore dose, with significantly different survivorship curves for the different doses. Moribund and dead fish exhibited characteristic ulcerous lesions at the injection site starting at 13 d p.i. None of the sham-injected control fish (0 zoospore treatment) died. The LD50 (lethal dose killing 50% of exposed menhaden) for inoculated fish was estimated at 9.7 zoospores; however, some fish receiving an estimated single zoospore developed infections that resulted in death. Menhaden were also challenged by aqueous exposure and confirmed that A. invadans was highly pathogenic by this more environmentally realistic route. Fish that were acclimated to culture conditions for 30 d, and presumably free of skin damage, then aqueously exposed to 100 zoospores ml(-1), exhibited 14% lesion prevalence with 11% mortality. Net-handled fish that were similarly infected had a significantly higher lesion prevalence (64%) and mortality (64%). Control fish developed no lesions and did not die. Scanning electron microscopy of fish skin indicated that zoospores adhered to intact epidermis, germinated and penetrated the epithelium with a germ tube. Our results indicate that A. invadans is a primary pathogen of menhaden and is able to cause disease at very low zoospore concentrations. PMID- 12747641 TI - Parasitic infections in freshwater ornamental fish in Sri Lanka. AB - A total of 1520 ornamental fish of 13 species from 26 export farms in Sri Lanka were collected between October 1999 and March 2000 and examined for parasites. Fish species examined were guppy Poecilia reticulata, goldfish Carassius auratus, platy Xiphophorus maculatus, molly Poecilia sphenops, angel Pterophyllum scalare, swordtail Xiphophorus helleri, tetras Hyphessobrycon species, barbs Capeota and Puntius spp., gourami Colisa sp., carp Cyprinus carpio, fighters Betta spelendens and others (Brachydanio and Astronotus spp.). Nine species of monogenean trematodes (Dactylogyrus extensus, Dactylogyrus cf. extensus, D. vastator, Dactylogyrus cf. vastator Dactylogyrus spp., Gyrodactylus turnbulli, G. katherineri, Gyrodactylus cf. katherineri, Gyrodactylus spp.), 7 protozoan species (Trichodina nigra, Trichodina spp., Tetrahymena corlissi, T. pyriformis, Ichthyophthirius multifiliis, Ichthyobodo necator, Piscinoodinium spp.), 3 species of copepod arthropods (Lernaea cyprinacea, Ergasilus ceylonensis, Argulus foliaceus), 1 metacercarial stage of a digenean trematode (Centrocestus spp.) and 1 nematode (Capillaria spp.) were identified. Parasites were found in fish from 23 of the 26 farms with an overall prevalence of parasitism in 45.3% of fish. The variation in farm prevalence among different parasites was significant (p < 0.01). Fish infection rates with monogenean trematodes, protozoans, copepod crustaceans, digenean trematodes and nematodes were 28.3, 18.4, 4.8, 0.8 and 0.4%, respectively. In all, 50 out of 590 (50/590) guppies were infected with Tetrahymena, compared with 13/930 for all other species, which is a statistically significant result (p < 0.01). Similarly, 13/44 and 18/44 carp were infected with Argulus foliaceus and Lernaea cyprinacea, compared with 7/1476 and 15/1476, respectively, for all other species combined (p < 0.01). Capillaria spp. was found only in guppies (4/590) and angel fish (3/92) while Centrocestus spp. was found in goldfish (12/153) only. PMID- 12747640 TI - Characterization of Cryptocaryon irritans, a parasite isolated from marine fishes in Taiwan. AB - The ciliated protozoan parasite Cryptocaryon irritans infecting marine fishes in Taiwan is described. Developmental characteristics and sequences of the ribosomal DNA regions such as part of 18 S, the entire first internal transcribed spacer, and part of 5.8 S of various Taiwan isolates of C. irritans were investigated. A total of 5 isolates was obtained from different fish-host species and localities, the majority from cultured fish species. C. irritans from Taiwan is able to shift its developmental characteristics, i.e. from non-adherent to adherent tomonts, from individualistic to aggregate-forming tomonts, from infection of the gills only to infection of the gills and body. Thus, it is not possible to classify strains of C. irritans on the basis of these parameters. Premature tomonts that developed from dead fishes were able to produce theronts that could infect fish host. Isolates from Pingtung and the USA had identical nucleotide sequences while an isolate from Malaysia was identical to an Israel isolate. Percentage variation among pairs of Taiwan isolates showed a higher degree of variation than isolate sequences listed in GenBank. Sequence analysis revealed highly aberrant isolates in Taiwan, and a phylogenetic tree distinguished a marine and a low-salinity variant. C. irritans from marine fishes in Taiwan, therefore, display some characteristics not previously reported. Since manipulation of salinity in brackishwater ponds and marine cage sites is not feasible, there is a need to develop new strategies for the control and prevention of cryptocaryoniasis. PMID- 12747642 TI - Isolation and 18S ribosomal DNA gene sequences of Marteilioides chungmuensis (Paramyxea), an ovarian parasite of the Pacific oyster Crassostrea gigas. AB - To develop sensitive detection techniques with the aim of elucidating the life cycle of Marteilioides chungmuensis, an intracellular paramyxean infecting the ovary of the Pacific oyster Crassostrea gigas, we isolated the parasite at the sporont stage from infected oysters using a freeze-thaw procedure at -20 degrees C and differential centrifugations in discontinuous sucrose and Percoll gradients. DNA was extracted from the isolated sporonts, and a PCR amplicon of 18S small subunit ribosomal RNA gene DNA was partially sequenced. In situ hybridization using 3 parasite-specific probes designed from the obtained sequence successfully detected parasite cells in infected oysters, and confirmed that the sequenced DNA was derived from M. chungmuensis. PMID- 12747643 TI - Detection of Myxobolus rotundus (Myxozoa: Myxosporea) in skin mucus of crucian carp Carassius auratus auratus using a monoclonal antibody. AB - Diagnosis of myxosporean Myxobolus rotundus infection was conducted by examining skin mucus from the infected crucian carp Carassius auratus auratus with a monoclonal antibody, MAb 2D12, raised previously against the parasite. A positive reaction was observed in skin mucus collected from infected fish, and spores and pre-spore stages of the parasite were identified by the MAb 2D12. It was also demonstrated that M. rotundus infection can be successfully detected by a simple method, enzyme-linked immunosorbent assay (ELISA), and that skin mucus collected from infected fish skin had a significantly higher optical density (OD) value than that from uninfected fish. PMID- 12747644 TI - Lens epithelial cells express CD95 and CD95 ligand treatment induces cell death and DNA fragmentation in vitro. AB - PURPOSE: Despite advances in intraocular lens design and material, posterior capsule opacification remains one of the major problems in modern cataract surgery. Therefore, the use of antiproliferative agents has been advocated. CD95 ligand (CD95L, Fas, Apo-1) is a death ligand that triggers apoptosis in susceptible target cells. Apoptosis allows for the safe disposal of cells without damaging the surrounding tissue. The goal of this study was to characterize and evaluate CD95L-induced cell death in cultured lens epithelial cells (LEC). METHODS: Expression of CD95 in untreated porcine LEC was investigated by flow cytometry. Cell death after CD95L or CD95 agonistic antibody treatment was assessed by crystal violet assay and DNA fragmentation was measured by comet assay. RESULTS: The presence of CD95 was observed in LEC. CD95L treatment resulted in a time--and concentration-dependent killing of LEC, which was synergistically enhanced by the addition of cyclohexamide. CD95L treatment induced DNA fragmentation. CONCLUSIONS: The present study confirms the use of apoptosis-inducing CD95L in the inhibition of LEC proliferation. Further studies are needed before clinical application of CD95L to inhibit posterior capsule opacification will be feasible. PMID- 12747645 TI - Biopsy of the conjunctiva in dry eye patients establishes a correlation between squamous metaplasia and dry eye clinical severity. AB - PURPOSE: The aim of this work was to evaluate the different grades of squamous metaplasia of the conjunctiva during the clinical course of dry eye syndrome, detecting the most characteristic morphological and morphometric changes by biopsy in order to provide a diagnostic classification. METHODS: The conjunctiva was studied under light microscopy by conventional histological methods and by morphometric analysis in 165 patients and in 33 controls. Patients were classified according to the Schirmer 1 test, break-up time, rose Bengal staining, osmolarity and impression cytology. The epithelium and connective tissue, with their different cells and other structures, were studied. RESULTS: The conjunctiva in dry eye patients showed progressive stratification, hyperplasia, hypertrophy and cellular flattening, with loss of goblet cell density and mucous layer. We found five pathological grades of squamous metaplasia and one normal grade. Clear nuclear alterations (indentation and binucleation) were found in the early grades of dry eye syndrome, but pyknotic nuclei and anucleated cells were only seen in the most severe grades. The smallest epithelial cells were found in the control group and their size increased with the severity of the dry eye syndrome. From the earliest stages to the most severe cases, increases in cellular separation were observed. There was also an increase in the number of inflammatory cells. Blood and lymphatic vessels showed alterations only in the most severe cases. CONCLUSIONS: This is the first grading system proposed for biopsy evaluation of the ocular surface in dry eye patients. These morphological and morphometric studies alone were able, even in the earliest phases of dry eye, to detect the squamous metaplasia that progresses from the surface of the epithelium to the connective tissue. This degenerative or adaptative cellular process was characterized mainly by marked increases in the stratification, epithelial cellular size and a general loss of goblet cells. PMID- 12747646 TI - Optical biometry before and after excimer laser epithelial keratomileusis (LASEK) for myopia. AB - PURPOSE: In order to select the correct intraocular lens (IOL) for implantation, it is important to measure the eye length (biometry). The IOL Master from Zeiss Humphry is frequently used for such measurements. Because this instrument employs an optical method, any irregularities on the corneal surface or any disturbances in corneal transparency could lead to mistakes in the measurements. The aim of this study was to determine whether eye length measurements obtained at the University Eye Clinic Regensburg, Germany with the IOL Master before and after excimer laser epithelial keratomileusis (LASEK) show any changes. METHODS: Axial length was measured on 20 myopic eyes (-2.75 to -8.00 diopters) before and one month after LASEK using the IOL Master. RESULTS: The mean pre-operative eye length was 25.46 mm (SD +/- 1.03) and the post-operative mean length was 25.38 mm (SD +/- 0.99). There was a strong correlation between the pre- and post-operative eye lengths (Pearson correlation coefficient 0.998). CONCLUSIONS: Although LASEK can lead to increased light scattering due to irregularities of the corneal surface and changes in corneal transparency, there is no difference in biometry pre- and post-operatively. PMID- 12747647 TI - Peripheral vasospasm and nocturnal blood pressure dipping--two distinct risk factors for glaucomatous damage? AB - PURPOSE: To evaluate the relationship between peripheral vasospasm and circadian blood pressure rhythm in patients with primary open angle glaucoma (POAG). METHODS: Nail-fold capillaroscopy, combined with a cold provocation test, and 24 hour blood pressure monitoring was carried out in 130 patients with POAG (M:F 58:72; mean age 60 +/- 14 years), 99 with high-tension glaucoma (HTG) and 31 with normal-tension glaucoma (NTG). Peripheral blood flow parameters were compared for patients with a nocturnal fall in mean systemic blood pressure (MBP) of less than 10% (non-dippers), patients with a nighttime MBP fall of 10-20% (dippers), and patients with a nighttime MBP fall of more than 20% (over-dippers). RESULTS: Patients with POAG showed a significantly lower blood flow velocity both at baseline (p < 0.01) and after cold provocation (p < 0.02) and a significantly higher percentage of cold-induced blood-flow standstill (p < 0.0001) in the nail fold capillaroscopy than normal controls. The numbers of non-dippers (50), dippers (66) and over-dippers (14) did not differ between the HTG and NTG group. There were no significant differences between non-dippers, dippers, and over dippers in peripheral blood flow parameters. CONCLUSIONS: Our findings indicate that vasospasm and low blood pressure may be distinct risk factors for glaucomatous damage. It also appears that screening for vascular dysregulation and systemic hypotension should not be restricted to NTG patients alone. PMID- 12747648 TI - Correlation between confocal scanning laser ophthalmoscopy and scanning laser polarimetry in open angle glaucoma. AB - PURPOSE: To correlate the findings of confocal scanning laser ophthalmoscopy and scanning laser polarimetry in diagnosed cases of glaucoma with established visual field defects. METHODS: A total of 53 diagnosed cases of primary open angle glaucoma that had at least two recorded of IOP more than 21 mm Hg on Goldmann applanation tonometry, open angles on gonioscopy and glaucomatous visual field defects on automated perimetry, were examined by confocal scanning laser ophthalmoscopy (HRT-II) and scanning laser polarimetry (GDx-NFA), in random order. RESULTS: The number on GDx advanced analysis had a significant (p < 0.05) correlation with the rim area (r = -0.279; p = 0.043), cup area (r = 0.311; p = 0.023) and the vertical cup: disc ratio (r = 0.376; p = 0.006). The correlation between HRT-II stereometric parameters and GDx advanced analysis parameters was significant (p < 0.05) for more parameters targeting the inferior pole of the disc than the superior pole. Numerically, the worst values of GDx parameters were associated with a worse result on Moorfields regression analysis, but there was poor agreement between the diagnostic labels like within or outside normal limits as obtained on GDx and HRT-II. CONCLUSIONS: Nerve fiber loss as detected on GDx correlates well with topographic optic nerve head changes as measured with the HRT-II. However, automated diagnosis on the two machines showed poor agreement. PMID- 12747649 TI - Changes in blood antioxidants and several lipid peroxidation products in women with age-related macular degeneration. AB - PURPOSE: The aim of this study was to evaluate the ferric reducing ability of plasma (FRAP), selected enzymatic and non-enzymatic components of the antioxidative system, and the intensity of peroxidative processes in the blood of patients with age-related macular degeneration (AMD). METHODS: In the peripheral blood, we evaluated FRAP; concentrations of vitamins C, A, and E; and of thiols. We assayed the activity of enzymatic components of the antioxidative system superoxide dismutase, catalase, ceruloplasmin and the concentration of reduced glutathione as an indicator of glutathione peroxidase activity. In order to determine the intensity of lipid peroxidation, we measured the concentrations of malondialdehyde and hydroxyalkenales (MDA-HNA) and conjugated diens (CD). RESULTS: We found a significant increase in FRAP in patients with AMD compared with the control group. The average concentrations of vitamins A and C were low and vitamins E and GSH were significantly higher in AMD than in the control group. The activity of almost all the antioxidative enzymes was high. We found a significant increase in MDA-HNA but no difference in CD. CONCLUSIONS: The significantly higher concentration of lipid peroxidation products in patients with AMD indicates an important pathogenic role of oxido-reduction disturbance. The high FRAP concentration may be one of the protective mechanisms in oxidation stress. The adaptive increase of the antioxidant barrier mostly involves the enzymatic components. PMID- 12747650 TI - Bottle-cork injury to the eye: a review of 13 cases. AB - PURPOSE: To analyze the anatomic and functional consequences of wine-cork injury to the eye in relation to the patient's age and the type of cork and wine. METHODS: We retrospectively studied 13 patients, six women and seven men, presenting to our department with bottle-cork injury to the eye between January 1999 and June 2001. RESULTS: All patients presented with closed-globe injury according to Kuhn et al's classification. All the cases were injured by bottle corks from sparkling wine: white in ten cases and red in three. Mean visual acuity at admission was 20/100 (range, hand motion to 20/20). The most frequent early injury was anterior chamber hyphema (84.6%), followed by corneal injury (62.2%), ocular hypertension (46.1%), lens subluxation (30.8%), traumatic cataract (23.1%), and post-traumatic retinal edema (23.1%). Mean final visual acuity was 20/25; the follow-up ranged from 3 to 29 months, averaging 16.1 months. Late complications were as follows: pupil motility anomalies (38.5%), traumatic cataract (30.8%), iridodialysis (15.4%), traumatic optic neuropathy (7.7%), post-traumatic glaucoma (7.7%), and traumatic maculopathy (15.4%). Surgical treatment was necessary in two cases (15.4%). CONCLUSIONS: Bottle-cork eye injuries account for 10.8% of post-traumatic hospital admissions to our department. Most of them are due to sparkling white wine served at room temperature. There is no correlation between ocular injury and the eye-bottle distance or the type of cork. PMID- 12747652 TI - Primary vitrectomy for pseudophakic retinal detachment: a prospective non randomized study. AB - PURPOSE: To compare the anatomic and functional results of primary vitrectomy alone or vitrectomy-scleral buckling for pseudophakic retinal detachment (RD). Vitrectomy permits a detailed view of the peripheral retina, so otherwise undetectable holes or additional small lesions can be found which, left untreated, may cause a residual RD. PATIENTS AND METHODS: Twenty-four consecutive pseudophakic eyes with pseudophakic RD were operated by vitrectomy and encircling band (Group I) and 27 additional cases were operated on by vitrectomy alone (Group II). Internal subretinal fluid drainage, using liquid perfluoro-n-octane, endolaser, and/or cryocoagulation and fluid-air exchange with SF6 20%, was applied in all cases. Preoperative findings and intraoperative and postoperative complications as well as final results were analyzed. RESULTS: Preoperatively undetected retinal holes were identified in 7 of the 51 eyes and additional retinal holes were found in 21. The mean follow-up was 14 months for Group I and 11.5 months for Group II. The retina was successfully reattached with a single operation in 22 of 24 eyes (92%) in Group I. One eye had a recurrence of RD due to an unsuccessfully treated preexisting retinal tear. Proliferative vitreoretinopathy (PVR) was observed in one case with recurrence of RD. In both cases, a second operation achieved retinal reattachment. In Group II, the retinas were attached with a single operation and visual acuity improved by an average of four or more lines in 62.5% of the vitrectomy-buckling group and in 55.5% of the vitrectomy group. The most frequent complication was a transient hypertony, in 21 cases. CONCLUSIONS: Surgical treatment of pseudophakic RD, combining vitrectomy and scleral buckling or vitrectomy alone, achieves very good anatomic and functional results. The advantages include more efficient detection of the peripheral detachment causing retinal lesions, and a lower redetachment rate than after extraocular surgery only. PMID- 12747651 TI - Intravitreal triamcinolone acetonide injection in blind painful eyes. Intraocular steroids as a treatment for blind painful red eyes. AB - PURPOSE: Phthisis bulbi results from different ocular conditions. We evaluated intravitreal triamcinolone acetonide as a treatment option in blind painful eyes. METHODS: Thirty-one patients with unilateral phthisis were randomly divided into two groups. Group A received 0.3 ml (12.5 mg) triamcinolone acetonide intravitreally and Group B 0.3 ml balanced salt solution after retrobulbar anesthesia. Treatment success was assessed by subjective response to pain and clinically by biomicroscopic evaluation of conjunctival congestion. Tonometry was done before and after treatment. Follow-ups were at 24 hours, 3 weeks, 3 and 6 months, and 1 and 2 years. RESULTS: Throughout the two-year follow-up, only two patients in Group A reported pain after the procedure and were retreated, one at week 4 and the other at week 7. Conjunctival congestion was significantly lower in Group A. Two patients with hypotony before treatment had normal tension after triamcinolone. All Group B patients were reinjected with triamcinolone because pain continued after balanced salt solution injection. No severe complications were found. CONCLUSIONS: Intravitreal triamcinolone acetonide may be effective and safe for treating blind painful eyes. PMID- 12747653 TI - Goldmann-Favre vitreoretinal degeneration. AB - PURPOSE: To describe a case of Goldmann-Favre vitreoretinal degeneration with typical clinical findings. METHODS: The case report of a healthy 47-year-old woman with typical clinical findings of Goldmann-Favre vitreoretinal degeneration is presented. She had complaints of reduced visual acuity and night blindness. Her parents were first cousins. RESULTS: Fundus examination revealed annular pigmentary degenerative changes, macular edema, and peripheral retinoschisis in both eyes. Electroretinography results were abnormal. Optical coherence tomography scans revealed hyporeflective spaces in the macular area and irregularities of the chorioretinal complex in the degenerative pigmentary areas. CONCLUSIONS: Typical fundus findings combined with night blindness and electroretinogram abnormalities permited the diagnosis of Goldmann-Favre vitreoretinal degeneration. PMID- 12747654 TI - Bilateral Acanthamoeba keratitis with late recurrence of the infection in a corneal graft: a case report. AB - PURPOSE: To report a case of bilateral Acanthamoeba keratitis with late, atypical recurrence after penetrating keratoplasty a chaud. METHODS: A 23-year-old contact lens wearer was treated for bilateral Acanthamoeba keratitis and underwent penetrating keratoplasty in the right eye for descemetocele with impending risk of perforation. The postoperative course was uneventful and topical steroids were combined with neomycin and propamidine. Two months after the operation in the right eye the patient presented with active infection in the left eye. One month later recurrence appeared in the right eye, as a central corneal infiltrate in the graft. RESULTS: Recurrences in both eyes were successfully treated with a combination of hexamidine and neomycin, and with polyhexamethylene biguanide respectively. The right eye was regrafted three months after the recurrence and penetrating keratoplasty was done two years later in the left eye. Both grafts were successful and remained clear. There has been no further recurrence in the long-term follow-up. CONCLUSIONS: Recurrence of Acanthamoeba keratitis after penetrating keratoplasty a chaud may occur even several months after the operation and the manifestation may be atypical. Current antiamoebal therapy was effective and regrafting in the quiet eye was successful. PMID- 12747655 TI - Bilateral central anterior stromal opacity of the cornea: dystrophy or degeneration? AB - PURPOSE: To describe a bilateral, central, oval corneal opacity not resembling any known corneal degeneration or dystrophy. METHODS: Ophthalmic examination, corneal topography, and pachymetry. RESULTS: A 30-year-old woman presented complaining of blurred vision. Biomicroscopic examination revealed bilateral, centrally located, oval, diffuse opacification of the anterior corneal stroma. Corneal topography showed slightly paracentral corneal flattening and irregular astigmatism in both eyes. Pachymetry indicated thinning of the central cornea in the left eye. No evidence of systemic disease was found. Family members had no corneal abnormalities. CONCLUSIONS: This unusual keratopathy closely resembles climatic proteoglycan stromal keratopathy. However, it is not clear whether the condition should be described as a dystrophy or degeneration. PMID- 12747656 TI - Trans-orbital intra-cranial air gun injury. AB - PURPOSE: To describe a case of trans-orbital intra-cranial air gun injury with a discussion on air gun related morbidity and mortality. METHODS: Case report and literature review. RESULTS: The air gun pellet travelled through the orbit without penetrating the globe. It passed into the middle cranial fossa through the superior orbital fissure and lodged in the temporal lobe. The patient was managed conservatively with antibiotics and antiepilectis. CONCLUSIONS: Air gun design has changed in recent years resulting in an increased morbidity and mortality. Stricter legislation on the sale and use of these weapons needs to be implemented. PMID- 12747657 TI - Transient third nerve palsy in a young patient with intracranial arteriovenous malformation. AB - PURPOSE: To describe a patient with transient third nerve palsy as the possible presenting sign of intracranial arteriovenous malformation. METHOD: Case report. RESULT: A 24-year-old female presented to ophthalmic casualty with sudden onset binocular diplopia and was diagnosed to have right sided partial third nerve palsy. Within 30 hours the third nerve palsy had recovered completely. A MRI scan and subsequent carotid angiogram revealed a large, high flow, trans-cortical Spetzler-Martin grade 4 arteriovenous malformation. The feeder vessel of the AVM originated from the right middle cerebral artery. Superficial venous drainage was via the superficial middle cerebral vein to the right transverse sinus. The deep venous drainage was via thalamostriate veins into markedly dilated internal cerebral vein and vein of Galen (Great cerebral vein). Venous reflux was noted around the midbrain from the vein of Galen. CONCLUSIONS: Transient third nerve palsy may rarely occur secondary to intracranial arteriovenous malformation. Ophthalmologists should consider neuroimaging in the investigations for transient cases of III nerve palsy in young patients. PMID- 12747658 TI - Tools for the diagnosis of equine respiratory disorders. AB - Respiratory disorders are among the most common problems leading horse owners to seek veterinary attention. Accurate diagnosis of these conditions allows for proper treatment to be instituted, much to the benefit of the patient and satisfaction of the client. As an introduction to this issue on equine respiratory disorders, we review some of the tools that are available to equine veterinarians for the diagnosis of respiratory disorders. Physical and endoscopic examination, radiology, diagnostic ultrasound, techniques for sampling the respiratory tract, hematology, blood gas analysis, respiratory mechanics, and some modern diagnostic tools are briefly covered. PMID- 12747659 TI - Lower respiratory problems of the neonate. AB - The newborn foal can experience problems of the lower respiratory tract that are unique to the neonate. The transition to extrauterine life usually occurs rapidly and in a coordinated manner, but problems associated with the peripartum period, including placentitis, dystocia, infection, and trauma, can result in conditions that compromise gas exchange in the newborn foal. This article reviews the normal transition and presents some of the problems seen in these small patients. PMID- 12747660 TI - Pneumonia in weanlings. AB - Lower respiratory tract infection is common in weanling- and suckling-aged animals. Increased susceptibility to disease in this age group can result from a delay in the establishment of a competent immune system and environmental factors, such as overcrowding, shipping, and sales. S zooepidemicus and R equi are the two most common bacterial isolates. S equi is primarily a disease of the lymph nodes and upper respiratory tract. Viral agents can compromise the natural defense mechanisms of the respiratory tract, resulting in secondary bacterial infections. The acute respiratory distress syndrome is one of unknown etiology and high mortality. PMID- 12747661 TI - Diagnosis of upper respiratory tract diseases in the performance horse. AB - Wastage of performance horses because of respiratory dysfunction is common. Appropriate identification of the disease is paramount for treatment recommendations. Diagnostic modalities for upper respiratory tract dysfunction include a thorough physical examination, radiographic evaluation when appropriate, and upper respiratory tract endoscopy. Anatomical deviations or structural are easily identified during resting evaluation, while exercise testing is often necessary to assess thedynamic properties of the upper airway. Utilizing the many diagnostic tools available allows the clinician to make an accurate diagnosis and recommend appropriate treatment. PMID- 12747662 TI - Recurrent airway obstruction--heaves. AB - Heaves, or recurrent airway obstruction (RAO), is a chronic respiratory disease featuring lower airway inflammation, bronchoconstriction, and mucus accumulation. Inhaled organic dusts and T helper 2 type immunologic reactions are involved in the complex pathophysiology of RAO. Clinical signs vary and alternate with remission periods. The diagnosis is often based on history and clinical examination in severe cases, but bronchoalveolar lavage may be useful for the detection of early cases. The most important aspect of treatment is to avoid exposure to allergens. Corticosteroids may be administered systemically or by inhalation in combination with bronchodilators and environmental control. PMID- 12747663 TI - Exercise-induced pulmonary hemorrhage. AB - EIPH is a condition affecting virtually all horses during intense exercise worldwide. The hemorrhage originates from the pulmonary vasculature and is distributed predominantly bilaterally in the dorsocaudal lung lobes. As the condition progresses, the lung abnormalities extend cranially along the dorsal portions of the lung. An inflammatory response occurs in association with the hemorrhage and may contribute to the chronic sequela. Although conflicting opinions exist as to its affect on performance, it is a syndrome that is thought to increase in severity with age. The most commonly performed method to diagnose EIPH at the present time is endoscopy of the upper airway alone or in combination with tracheal wash analysis for the presence of erythrocytes and hemosiderophages. Because horses may not bleed to the same extent every time and the bleeding may originate from slightly different locations, these diagnostic procedures may not be extremely sensitive or quantitative. At this time, there is no treatment that is considered a panacea, and the currently allowed treatments have not proven to be effective in preventing EIPH. Future directions for therapeutic intervention may need to include limiting inflammatory responses to blood remaining within the lungs after EIPH. PMID- 12747665 TI - Diseases of the guttural pouches. AB - Guttural pouches are unique anatomic structures in Equidae. Conditions affecting the guttural pouches present with clinical signs that are not observed in other domestic species, and treatment of some of these conditions can be challenging. This article describes the anatomy and examination of the guttural pouches and reviews the presenting signs and treatment of the more common conditions affecting them, including empyema, tympany, mycosis, and stylohyoid arthropathy. PMID- 12747666 TI - Disorders of the pharynx. AB - Pharyngeal disorders are complex and difficult to treat. Disorders that lead to anatomic derangement, such as trauma and neoplasia, can significantly affect the function of this organ. Pharyngeal dysfunction can manifest as dysphagia, persistent palatal displacement, or exercise intolerance. Secondary complications are serious and life threatening and include aspiration pneumonia, weight loss, and death. Pharyngeal disorders that are only recognizable during strenuous exercise are difficult to diagnose and are treated with limited success, even though they are responsible for significant economic losses with performance animals. PMID- 12747664 TI - Lower airway diseases of the adult horse. AB - Lower airway problems of the adult horse are commonly encountered by the practitioner. Particularly susceptible populations include horses transported for any significant distance and young horses grouped together for training and/or competition. This article presents some of the commonly encountered problems of this patient population, including bacterial pneumonia/pleuropneumonia and influenza, and some uncommon ones, including pulmonary edema, pneumothorax/hemothorax, and acuterespiratory distress syndrome. Information is presented that should allow the practitioner to diagnose these problems accurately and initiate rational treatment plans. PMID- 12747667 TI - Disorders of the larynx. AB - The upper respiratory tract is a frequent cause of exercise intolerance in horses, particularly in racing horses. There are a myriad of laryngeal abnormalities that may restrict airflow at the rima glottidis. Careful endoscopic examination is a crucial part of the examination of any racing horse suffering from poor performance. There has recently been interest in spectrum analysis of respiratory sounds. It has been determined that laryngeal hemiplegia and dorsal displacement of the soft palate have unique sound patterns. Therefore, spectrum analysis of respiratory sounds may prove to be useful in the diagnosis of laryngeal disorders in horses. Accurate diagnosis and appropriate surgical intervention are necessary to provide the horse the best chance of returning to its full athletic potential. PMID- 12747669 TI - Sinus disease. AB - The diagnosis and treatment of diseases of the paranasal sinuses and conchae of horses are complicated by the large size of these structures, their complex anatomy, difficulties of access to them, and the advanced state of many diseases before diagnosis is made. Improved diagnostic methods include nuclear scintigraphy, computed tomography, and direct endoscopy of the sinuses. Treatment of some sinus diseases can be enhanced by access through direct sinus endoscopy for focal lesions, use of large frontal bone flaps for diffuse sinus lesions, standing surgery through a frontal flap for a variety of sinus disorders, and intralesional formalin for progressive ethmoidal hematomas. PMID- 12747668 TI - Laryngeal reinnervation in the horse. AB - Left laryngeal hemiplegia is a frustrating condition for the equine athlete and equine veterinarian. Treatment for the past 30 years has centered on the prosthetic laryngoplasty ("tie-back") with or without ventriculectomy. Laryngeal reinnervation has been used successfully in people and has been shown experimentally to benefit affected horses. This article reviews equine laryngeal reinnervation using the nerve muscle pedicle graft and describes the surgical technique, its complications, and the follow-up in 146 cases treated over the past 10 years. Also discussed is ongoing research into stimulation studies to improve the success of equine laryngeal reinnervation. PMID- 12747670 TI - The use of lasers for treatment of upper respiratory tract disorders. AB - Lasers have become important tools for the equine surgeon in the treatment of upper respiratory tract disease in the horse. Multiple wavelengths and delivery systems are available. Indications for the use of lasers in the upper respiratory tract primarily include minimally invasive procedures not possible with conventional surgical instrumentation. New applications for the use of lasers to treat upper respiratory disease are likely to evolve with the development and introduction of new wavelengths and delivery systems. PMID- 12747671 TI - Bioresorbable polymeric stents: current status and future promise. AB - Metal stents and, more recently, polymer-coated metal stents are used to stabilize dissections, eliminate vessel recoil, and guide remodeling after balloon angioplasty and other treatments for arterial disease. Bioresorbable polymeric stents are being developed to improve the biocompatibility and the drug reservoir capacity of metal stents, and to offer a transient alternative to the permanent metallic stent implant. Following a brief review of metal stent technology, the emerging class of expandable, bioresorbable polymeric stents is described, with emphasis on developments in the authors' laboratory. PMID- 12747672 TI - Characterization of poly(ethylene oxide) brushes on glass surfaces and adhesion of Staphylococcus epidermidis. AB - Poly(ethylene oxide) brushes have been covalently bound to glass surfaces and their presence was demonstrated by an increase in water contact angles from fully wettable on glass to advancing contact angles of 54 degrees, with a hysteresis of 32 degrees. In addition, electrophoretic mobilities of glass and brush-coated glass were determined using streaming potential measurements. The dependence of the electrophoretic mobilities on the ionic strength was analyzed in terms of a softlayer model, yielding an electrophoretic softness and fixed charge density of the layer. Brush-coated glass could be distinguished from glass by a 2-3-fold decrease in fixed charge density, while both surfaces were about equally soft. Adhesion of Staphylococcus epidermidis HBH276 to glass in a parallel plate flow chamber was extremely high and after 4 h, 19.0 x 10(6) bacteria were adhering per cm2. In contrast, the organisms did not adhere to brush-coated glass, with numbers below the detection limit, i.e. 0.1 x 10(6) per cm2. These results attest to the great potential of polymer brushes in preventing bacterial adhesion to surfaces. PMID- 12747673 TI - Tissue-culture surfaces with mixtures of aminated and fluorinated functional groups. Part 1. Synthesis and characterization. AB - Surface chemistry of culture dishes can have profound effects on the phenotype of cultured cells. In the present study, chemisorption from aqueous, binary mixtures of organosilanes onto borosilicate glass created surfaces bearing diamine groups (N2), trifluoropropyl groups (F3) and mixtures of the two. Composition of N2-F3 surfaces was controlled by the ratio of monomers in the silanization bath, as confirmed by electron spectroscopy for chemical analysis and by conjugation of surface amines with fluorescein-5-isothiocyanate. Atomic-force microscopy revealed that silanized surfaces are patchy, though their root-mean-square roughnesses do not differ significantly from that of smooth glass (0.3 nm). Surfaces richest in diamine residues were the most hydrophilic, with advancing water-contact angles < or = 90 degrees. The accompanying paper (the next article in this issue) describes the effects of these surface chemistries on the phenotype of transgenic insulinoma cells in vitro. We conclude that chemisorption from the N2-F3 system provides a simple, one-pot method for tailoring the chemistry of glass culture surfaces. PMID- 12747674 TI - Tissue-culture surfaces with mixtures of aminated and fluorinated functional groups. Part 2. Growth and function of transgenic rat insulinoma cells (betaG I/17). AB - Interactions of transplantable cells with synthetic polymers can influence the function of biohybrid artificial organs. This study explored growth and secretion of human insulin by betaG I/17 cells cultured on surfaces bearing diamine groups (N2), trifluoropropyl groups (F3) and mixtures of the two. Cells cultured on high F3 and high-N2 surfaces spread well, grew rapidly and produced >1.8 mol lactate per mol glucose consumed, closely resembling cells grown on the permissive control, glass. On one mixed surface, with a molar ratio of 33 N2 groups:67 F3 groups, cells had a lower lactate/glucose ratio, adopted a rounded form, grew slowly and were quick to form emergent aggregates, similar to cultures on the inhibitory control, untreated polystyrene. Cultures on surfaces with higher F3 content secreted the most insulin and, in the case of the highest-F3 surface, showed improved responsiveness to secretagogues. Hormone secretion was roughly 50% greater when cells were grown on F3 surfaces conditioned by earlier cultures of betaG I/17. Incubation of conditioned surfaces with high concentrations of a polyclonal anti-laminin serum prior to re-plating partially abolished this improvement in secretory function. Polymers bearing trifluoropropyl groups appear to be attractive candidates for use in the artificial endocrine pancreas. Surface coatings that include laminin might promote function of transgenic insulinoma cells in vitro and in vivo. PMID- 12747675 TI - Effect of physiological temperature on the mechanical properties and network structure of biodegradable poly(propylene fumarate)-based networks. AB - Poly(propylene fumarate) (PPF)-based networks have exhibited increases in mechanical properties during their initial stages of degradation. This study was designed to investigate whether physiological temperatures are the source of this reinforcing behavior by influencing the formation of additional crosslinks within the network. Utilizing a model PPF network formed with the crosslinking agent poly(propylene fumarate)-diacrylate (PPF-DA), cylindrical specimens were stored in an inert environment and conditioned at -20 and 37 degrees C while their mechanical properties and network structure were monitored over a six week period. The PPF/PPF-DA specimens exposed to physiological temperatures showed an increase in compressive modulus from 1674 +/- 88 to 2059 +/- 75 MPa. The double bond conversion improved as well, from 64 +/- 1 to 70 +/- 1%, indicating that crosslinks were being formed in the network. The additional reactivity occurred exclusively with unreacted fumarate bonds. PPF/PPF-DA networks stored at -20 degrees C showed no changes in mechanical properties; however, they increased when subsequently conditioned at 37 degrees C. The results were used to explain that PPF-based networks undergo a biphasic degradation behavior due to the competing hydrolytic degradation and thermal induced crosslinking. In addition, heat treating the networks at higher temperatures can be utilized as a means to further reinforce PPF-based materials. PMID- 12747676 TI - Chemically-bound nerve growth factor for neural tissue engineering applications. AB - In order to promote regeneration after spinal cord injury, growth factors have been applied in vivo to rescue ailing neurons and provide a path finding signal for regenerating neurites. We previously demonstrated that soluble growth factor concentration gradients can guide axons over long distances, but this model is inherently limited to in vitro applications. To translate the use of growth factor gradients to an implantible device for in vivo studies, we developed a photochemical method to bind nerve growth factor (NGF) to microporous poly(2 hydroxyethylmethacrylate) (PHEMA) gels and tested bioactivity in vitro. A cell adhesive photoreactive poly(allylamine) (PAA) was synthesized and characterized. This photoreactive PAA was applied to the surface of the PHEMA gels to provide both a cell adhesive layer and a photoreactive handle for further NGF immobilization. Using a direct ELISA technique, the amount of NGF immobilized on the surface of PHEMA after UV exposure was determined to be 5.65 +/- 0.82 ng/cm2 or 3.4% of the originally applied NGF. A cell-based assay was performed to determine the bioactivity of the immobilized NGF. Using pheochromocytoma (PC-12) cells, 30 +/- 7% of the cell population responded to bound NGF, a response statistically similar to that of cells cultured on collagen in the presence of 40 ng/ml soluble NGF of 39 +/- 12%. These results demonstrate that PHEMA with photochemically bound NGF is bioactive. This photochemical technique may be useful to spatially control the amount of NGF bound to PHEMA using light and thus build a stable concentration gradient. PMID- 12747677 TI - Improved detection of Salmonella spp. in foods by fluorescent in situ hybridization with 23S rRNA probes: a comparison with conventional culture methods. AB - This report describes a new technique for the detection and identification of Salmonella species in food with the use of fluorescent in situ hybridization (FISH) with 23S rRNA-targeted oligonucleotide probes. Two species-specific 23S rRNA-targeted oligonucleotide probes (Sal-1 and Sal-3) were selected, and one (Sal-544) was newly designed. The relative specificities of these probes were compared with those of bacterial 23S rRNA sequences from the GenBank database and tested by in situ hybridization with bacterial cell smears of pure cultures. Fifty-one tested reference strains of Salmonella serovars belonging to subspecies I (enterica) hybridized with these probes. No cross-reactions with 46 other strains of the family Enterobacteriaceae or with another 14 bacterial strains from other families were observed. Storage of a Salmonella Panama test strain under various environmental conditions (2, 5, and 15% NaC1; -20 degrees C, 4 degrees C, and room temperature; pHs of 3.3 to 7.4) did not adversely affect the FISH method. No matrix effects were observed with 18 different kinds of foods. FISH was able to detect Salmonella spp. in 52 (probe Sal-1), 56 (probe Sal-3), and 35 (probe Sal-544) of 225 naturally contaminated food samples after 16 h of incubation in a preenrichment broth. When conventional culture and detection methods were used, Salmonella could be isolated from only 30 of these 225 samples. In contrast, FISH failed to identify Salmonella in only two of the culture-positive samples when Sal-1 and Sal-3 were used and in only three of the culture-positive samples when Sal-544 was used. PMID- 12747678 TI - Comparative analysis of acid resistance between susceptible and multi antimicrobial-resistant Salmonella strains cultured under stationary-phase acid tolerance-inducing and noninducing conditions. AB - This study compared acid resistance levels among five antimicrobial-susceptible strains of Salmonella and five strains that were simultaneously resistant to a minimum of six antimicrobial agents. The induction of a stationary-phase acid tolerance response (ATR) was attempted by both transient low-pH acid shock and acid adaptation. For acid shock induction, strains were grown for 18 h in minimal E medium containing 0.4% glucose (EG medium) and exposed to sublethal acid stress (pH 4.3) for 2 h, and subsequently, both shocked and nonshocked cultures were acid challenged (pH 3.0) for 4 h. Acid adaptation was achieved by growing strains for 18 h in tryptic soy broth containing 1.0% glucose (TSB+G), while nonadapted cultures were grown for 18 h in glucose-free tryptic soy broth (TSB-G). Acid adapted and nonadapted inocula were acid challenged (pH 2.3) for 4 h. Initial (0 h) mean populations of nonchallenged Salmonella were 8.5 to 8.7, 8.4 to 8.8, and 8.2 to 8.3 log CFU/ml for strains grown in EG medium, TSB-G, and TSB+G, respectively. After 4 h of acid challenge, mean populations were 3.0 to 4.8 and 2.5 to 3.7 log CFU/ml for previously acid-shocked susceptible and resistant strains, respectively, while corresponding counts for nonshocked strains were 4.3 to 5.5 log CFU/ml and 3.9 to 4.9 log CFU/ml. Following 4 h of acid exposure, acid adapted cultures of susceptible and resistant strains had mean populations of 6.1 to 6.4 log CFU/ml and 6.4 to 6.6 log CFU/ml, respectively, while corresponding counts for nonadapted cultures were 1.9 to 2.1 log CFU/ml and 1.8 to 2.0 log CFU/ml, respectively. A low-pH-inducible ATR was not achieved through transient acid shock, while an ATR was evident following acid adaptation, as adapted populations were 4.2 to 4.8 log units larger than nonadapted populations following acid exposure. Although some strain-dependent variations in acid resistance were observed, results from this study suggest no association between susceptibility to antimicrobial agents and the ability of the Salmonella strains evaluated to survive low-pH stress. PMID- 12747680 TI - Prevalence of Escherichia coli O157:H7 and performance by beef feedlot cattle given Lactobacillus direct-fed microbials. AB - Fecal shedding of Escherichia coli O157:H7, the prevalence of Escherichia coli O157:H7 in pens and on carcasses and hides, and cattle performance as a result of daily dietary supplementation with Lactobacillus-based direct-fed microbials (DFMs) were evaluated in a feeding trial involving 180 beef steers. Steers were evaluated for shedding of E. coli O157:H7 by an immunomagnetic separation technique on arrival at the feedlot, just before treatment with the DFMs, and every 14 days thereafter until slaughter. Composite pen fecal samples were collected every 14 days (alternating weeks with animal testing), and prevalence on hides and carcasses at slaughter was also evaluated. Feedlot performance (body weight gain and feed intake) was measured for the period during which the DFMs were fed. Gain efficiency was calculated as the ratio of weight gain to feed intake. Lactobacillus acidophilus NPC 747 decreased (P < 0.01) the shedding of E. coli O157:H7 in the feces of individual cattle during the feeding period. E. coli O157:H7 was approximately twice as likely to be detected in control animal samples as in samples from animals receiving L. acidophilus NPC 747. In addition, DFM supplementation decreased (P < 0.05) the number of E. coli O157:H7-positive hide samples at harvest and the number of pens testing positive for the pathogen. Body weight gains (on a live or carcass basis) and feed intakes during the DFM supplementation period did not differ among treatments. Gain efficiencies on a live-weight basis did not differ among treatments, but carcass-based gain/feed ratios tended (P < 0.06) to be better for animals receiving the two DFM treatments than for control animals. The results of this study suggest that the feeding of a Lactobacillus-based DFM to cattle will decrease, but not eliminate, fecal shedding of E. coli O157:H7, as well as contamination on hides, without detrimental effects on performance. PMID- 12747679 TI - Reducing Salmonella on apples with wash practices commonly used by consumers. AB - The efficacy levels of practices used by consumers to wash smooth-surface fruits and vegetables were compared. Golden Delicious apples were spot inoculated near the blossom end with 50 microl of a cocktail of six serotypes of Salmonella enterica (with a total inoculum level of approximately 10(9) CFU per apple). The inoculum was dried for 1.5 h, and apples were either treated immediately or held for 24 h prior to treatment. Treatments included wetting with approximately 5 ml of water, vinegar (5% acidity), or a 200-ppm chlorine solution, rubbing for 5 or 30 s, rinsing with 200 to 600 ml of 24 or 43 degrees C water, and drying with a sterile paper towel. Residual populations of Salmonella were determined by rubbing the treated apple for 30 s in 20 ml of Dey-Engley neutralizing broth and plating on tryptic soy agar and bismuth sulfite agar. Rubbing treatments carried out for 5 and 30 s both resulted in a significant reduction in Salmonella populations (1 log10 CFU per apple) relative to populations on samples held for 30 s. A 5-s rub followed by a 200-ml flowing-water rinse reduced populations by 3 log10 CFU per apple. No further decrease in population was obtained by rinsing with 400 or 600 ml of water. Increasing the rinse water temperature to 43 degrees C did not significantly improve microbial removal. Drying the apple with a sterile paper towel resulted in an additional decrease of approximately 0.4 log10 CFU per apple. A reduction of 3.2 log10 CFU was achieved with a combination of wetting with water, rubbing for 5 s, rinsing with 200 ml of water, and drying with a paper towel for apples inoculated just prior to or 24 h before treatment. Reductions obtained for apples treated with 5% vinegar and with a 200-ppm chlorine solution were significantly larger (2.1 to 3.2 log10 CFU per apple, respectively) than those achieved with water. PMID- 12747681 TI - Effects of pH, temperature, and pre-pulsed electric field treatment on pulsed electric field and heat inactivation of Escherichia coli O157:H7. AB - This investigation was undertaken to study the inactivation of Escherichia coli O157:H7 by pulsed electric field (PEF) treatment and heat treatment after exposure to different stresses. E. coli O157:H7 cells exposed to different pHs (3.6, 5.2, and 7.0 for 6 h). different temperatures (4, 35, and 40 degrees C for 6 h), and different pre-PEF treatments (10, 15, and 20 kV/cm) were treated with PEFs (20, 25, and 30 kV/cm) or heat (60 degrees C for 3 min). The results of these experiments demonstrated that a pH of 3.6 and temperatures of 4 and 40 degrees C caused significant decreases in the inactivation of E. coli O157:H7 by PEF treatment and heat treatment (P < 0.05). Pre-PEF treatments, pHs of 5.2 and 7.0, and a temperature of 35 degrees C, on the other hand, did not result in any resistance of E. coli O157:H7 cells to inactivation by PEF treatment and heat treatment (P > 0.05). PMID- 12747682 TI - Irradiation D-values for Escherichia coli O157:H7 and Salmonella sp. on inoculated broccoli seeds and effects of irradiation on broccoli sprout keeping quality and seed viability. AB - Like alfalfa sprouts, broccoli sprouts can be a vehicle for bacterial pathogens, which can cause illness when they are consumed. The gamma irradiation process was used to reduce numbers of bacterial pathogens on broccoli sprouts and seeds, and the effect of this process on the seeds was studied. The irradiation destruct values for Salmonella sp. and for strains of Escherichia coli O157:H7 inoculated on broccoli seeds were determined. Results obtained in this study indicate that a dose of 2 kGy reduced total background counts for broccoli sprouts from 10(6) to 10(7) CFU/g to 10(4) to 10(5) CFU/g and increased the shelf life of the sprouts by 10 days. Yield ratio (wt/wt), germination percentage, sprout length, and thickness were measured to determine the effects of various irradiation doses on the broccoli seeds. Results show a decreased germination percentage at a dose level of 4 kGy, whereas the yield ratio (wt/wt), sprout length, and thickness decreased at the 2-kGy dose level. The radiation doses required to inactivate Salmonella sp. and strains of E. coli O157:H7 were higher than previously reported values. D-values, dose required for a 1-log reduction, for the nonvegetable and vegetable Salmonella sp. isolates were 0.74 and 1.10 kGy, respectively. The values for the nonvegetable and vegetable isolated strains of Escherichia coli O157:H7 were 1.43 and 1.11 kGy, respectively. With the irradiation process, a dose of up to 2 kGy can extend the shelf life of broccoli sprouts. A dose of > 2 kGy would have an adverse effect on the broccoli seed and decrease the yield of broccoli sprouts. PMID- 12747684 TI - Ozone treatment for reduction of Escherichia coli 0157:H7 and Salmonella serotype typhimurium on beef carcass surfaces. AB - The effectiveness of an aqueous ozone treatment in reducing Escherichia coli O157:H7 and Salmonella serotype Typhimurium on hot carcass surfaces was determined with the use of a model carcass spray cabinet. Carcass surface regions were removed from carcasses and inoculated with feces containing 10(6) to 10(7) CFU each of E. coli O157:H7 and Salmonella Typhimurium per g and were then exposed to a water wash or to a water wash followed by a sanitizing ozone treatment. Water washes were applied at 28 degrees C beginning at a pressure of 10 lb/in2 and gradually increasing to 400 lb/in2. Ozone treatment was carried out by spraying surfaces with an aqueous ozone solution (80 lb/in2 at 28 degrees C) containing 95 mg of ozone per liter. Pathogen reductions achieved with ozone treatment were not significantly different from those achieved with a water wash alone. In addition, ozone treatment did not reduce E. coli O157:H7 or Salmonella Typhimurium contamination that was spread over the carcass surface as a result of the water wash. Under the conditions of this study, the aqueous ozone treatment applied resulted in no significant improvement over a water wash in reducing pathogens on beef carcass surfaces. PMID- 12747683 TI - Chemical and irradiation treatments for killing Escherichia coli O157:H7 on alfalfa, radish, and mung bean seeds. AB - In this study, the effectiveness of dry-heat treatment in combination with chemical treatments (electrolyzed oxidizing [EO] water, califresh-S, 200 ppm of active chlorinated water) with and without sonication in eliminating Escherichia coli O157:H7 on laboratory-inoculated alfalfa, radish, and mung bean seeds was compared with that of dry-heat treatment in combination with irradiation treatment. The treatment of mung bean seeds with EO water in combination with sonication followed by a rinse with sterile distilled water resulted in reductions of approximately 4.0 log10 CFU of E. coli O157:H7 per g. whereas reductions of ca. 1.52 and 2.64 log10 CFU/g were obtained for radish and alfalfa seeds. The maximum reduction (3.70 log10 CFU/g) for mung bean seeds was achieved by treatment with califresh-S and chlorinated water (200 ppm) in combination with sonication and a rinse. The combination of dry heat, hot EO water treatment, and sonication was able to eliminate pathogen populations on mung bean seeds but was unable to eliminate the pathogen on radish and alfalfa seeds. Other chemical treatments used were effective in greatly reducing pathogen populations on radish and alfalfa seeds without compromising the quality of the sprouts, but these treatments did not result in the elimination of pathogens from radish and alfalfa seeds. Moreover, a combination of dry-heat and irradiation treatments was effective in eliminating E. coli O157:H7 on laboratory-inoculated alfalfa, radish, and mung bean seeds. An irradiation dose of 2.0 kGy in combination with dry heat eliminated E. coli O157:H7 completely from alfalfa and mung bean seeds, whereas a 2.5-kGy dose of irradiation was required to eliminate the pathogen completely from radish seeds. Dry heat in combination with irradiation doses of up to 2.0 kGy did not unacceptably decrease the germination percentage for alfalfa seeds or the length of alfalfa sprouts but did decrease the lengths of radish and mung bean sprouts. PMID- 12747685 TI - Escherichia coil O157 diversity with respect to survival during drying on concrete. AB - Shiga toxin (Stx)-producing Escherichia coli O157 isolates (n = 123) were divided into groups according to origin, genotype (pulsed-field gel electrophoresis [PFGE] type, or ribotype), type of Stx produced, or phage type (PT). The survival rate ([number of CFU after 24 h of drying/number of CFU before drying] x 100) for each isolate was determined in triplicate after drying on concrete for 24.0 h. The overall mean survival rate among the 123 E. coli O157 isolates studied was 22.9%, but there was a wide range of responses to drying on concrete, with a minimum of 1.2% and a maximum of 61.9% of the initial inocula being recovered after drying. Among the groups, those isolates that originated from cases of human disease were, on average, significantly more sensitive (P < 0.001) to drying (with a mean survival rate of 15.3%) than isolates from the other three sources (with mean survival rates of 27.7, 26.0, and 22.9% for meats, bovine or ovine feces, and bovine hides, respectively). When the isolates were grouped by genotype, three of the PFGE types were, on average, significantly more resistant to drying than two other PFGE types were, and similarly, significant differences in average resistance to drying between groups of E. coil O157 with different ribotypes were seen. There were no differences between the abilities of isolates producing different Stxs (Stx 1 or Stx 1 and Stx 2) to survive drying. E. coli O157 isolates of PT4, PT21/28, and PT32 survived drying on concrete better than groups of other PTs did. Since the E. coli O157 isolates had various abilities to survive drying on concrete, drying could contribute to a kind of E. coil O157 natural selection along the meat chain. This possibility may have significant meat safety implications if a range of E. coil O157 isolates are simultaneously exposed to drying at any point along the meat production chain. Those E. coil O157 isolates that are more able to survive drying could be more likely to pass farther along the meat chain and ultimately reach consumers. PMID- 12747686 TI - rP33 activates bacterial killing by chicken peripheral blood heterophils. AB - The protection of poultry from infection by Salmonella is of major concern with regard to human health because Salmonella is a common bacterial cause of foodborne diseases, and protection without the use of antibiotics is preferable in order to avoid possible complications involving antibiotic resistance. Salmonella immune lymphokine (SILK), produced by stimulated splenic T cells from Salmonella Enteritidis-immunized chickens, has been shown to confer protection against Salmonella infection on day-old chicks without the use of antibiotics. This protection results from the potentiation of an immune response following treatment with SILK. This study was undertaken to analyze a component of SILK, identified as P33, that is the product of the chicken mim-1 gene. A recombinant derivative expressing a domain of P33 (rP33) has been shown to be chemotactic for heterophils and is therefore instrumental in eliciting the immune response characteristic of SILK-induced protection against Salmonella infection in chicks. We report here that rP33 possesses the ability to activate antimicrobial responses from heterophils. The killing of Salmonella Enteritidis by heterophils was increased by in vitro treatment of the cells with rP33. Treatment with rP33 also stimulated the degranulation of heterophils but did not induce an oxidative burst or upregulate phagocytosis. These results indicate that P33 is an active component of SILK, conferring protection against Salmonella Enteritidis by augmenting the antimicrobial activities of heterophils. PMID- 12747687 TI - Serial disinfection with heat and chlorine to reduce microorganism populations on poultry transport containers. AB - A prototype system for the cleaning and decontamination of poultry transport containers was previously developed and evaluated as a means of eliminating foodborne pathogens entering poultry processing plants. While decontamination of the containers once with the use of either hot water (up to 70 degrees C) or sodium hypochlorite (up to 1,000 ppm) resulted in significant reductions in the numbers of coliforms and the elimination of small numbers of Salmonella, complete removal of pathogens was not attained. Therefore, the present study was conducted to determine whether repeated decontamination of the same containers could eliminate coliforms and Salmonella consistently. Individual five-tier containers consisting of galvanized steel frames and fiberglass floors were identified (n = 6) and decontaminated once per day for five consecutive days after being used to haul broilers from farms to the processing plant. Two types of containers were tested in this study: one had previously been used for broiler transportation, and the other had new floors. After each transport, the containers were first precleaned with a cleaning agent using a high-pressure jet (6,094 kPa) to remove debris and to loosen biofilms from surfaces. The containers were then immersed in an aqueous solution of 1,000 ppm of sodium hypochlorite at 70 degrees C for 2 min. Samples obtained from the container surfaces before and after each cleaning and decontamination were analyzed to obtain coliform and Salmonella counts. Coliforms were completely eliminated from both types of containers following one decontamination treatment. Because no Salmonella were detected on the containers, the effect of decontamination in the elimination of Salmonella was not determined. Similar treatments on five successive days also resulted in poultry transport containers that were essentially free of Salmonella and coliforms. This decontamination system involving a combination of heat and sodium hypochlorite can be used as a standard method for cleaning poultry transport containers in the poultry industry. It is recommended that such containers be cleaned after each use to avoid the potential risk of a buildup of significantly higher loads of pathogenic microorganisms and their biofilms. PMID- 12747688 TI - Comparison of recovery methods for freeze-injured Listeria monocytogenes, Salmonella Typhimurium, and Campylobacter coli in cell suspensions and associated with pork surfaces. AB - Cells injured as a result of freezing, heating, and acidification treatments may not grow during conventional microbiological procedures owing to the presence of selective agents, compounds, or dyes in the media, impairing the cell's ability to repair itself and grow. Injured cells can be recovered by combining selective and nonselective media into a single system. With such combinations, the diffusion of the selective compounds or dyes is controlled, allowing for the resuscitation of injured cells of interest while also inhibiting the growth of undesirable background microflora. In this study, Listeria monocytogenes, Salmonella Typhimurium, and Campylobacter coli suspended in buffer or associated with pork surfaces were subjected to a freeze-thaw cycle (-15 degrees C for 24 h, 4 degrees C for 4 h). Following treatments, freeze-injured cells were plated on appropriate media for the overlay (OV), thin agar layer (TAL), and Lutri plate (LP) recovery methods. The levels of L. monocytogenes and Salmonella Typhimurium recovered from cell suspensions and pork surfaces by the TAL, OV, and LP methods following freeze treatments were not statistically different (P > 0.05) from recovery levels associated with nonselective media. Conversely, levels of pathogens on selective media were significantly reduced compared with those for the other methods employed. The TAL method's recovery of C. coli was not significantly different from that achieved with the nonselective media. Overall, the results presented in this study demonstrate that the TAL method not only was easier to perform, but also allowed improved isolation of single colonies for further characterization. This study may provide researchers with better methods to determine the effectiveness of industry-employed chilling processes in reducing pathogenic bacteria associated with red meat surfaces. PMID- 12747689 TI - Predictive model for the combined effect of temperature, sodium lactate, and sodium diacetate on the heat resistance of Listeria monocytogenes in beef. AB - The effects of heating temperature (60 to 73.9 degrees C), sodium lactate (NaL; 0.0 to 4.8% [wt/wt]), and/or sodium diacetate (SDA; 0.0 to 0.25% [wt/wt]) and of the interactions of these factors on the heat resistance of a five-strain mixture of Listeria monocytogenes in 75% lean ground beef were examined. Thermal death times for L. monocytogenes in filtered stomacher bags in a circulating water bath were determined. The recovery medium was tryptic soy agar supplemented with 0.6% yeast extract and 1% sodium pyruvate. Decimal reduction times (D-values) were calculated by fitting a survival model to the data with a curve-fitting program. The D-values were analyzed by second-order response surface regression for temperature, NaL level, and SDA level. The D-values observed for beef with no NaL or SDA at 60, 65, 71.1, and 73.9 degrees C were 4.67, 0.72, 0.17, and 0.04 min, respectively. The addition of 4.8% NaL to beef increased heat resistance at all temperatures, with D-values ranging from 14.3 min at 60 degrees C to 0.13 min at 73.9 degrees C. Sodium diacetate interacted with NaL, thereby reducing the protective effect of NaL and rendering L. monocytogenes in beef less resistant to heat. A mathematical model describing the combined effect of temperature, NaL level, and SDA level on the thermal inactivation of L. monocytogenes was developed. This model can predict D-values for any combination of temperature, NaL level, and SDA level that is within the range of those tested. This predictive model will have substantial practical importance to processors of cooked meat, allowing them to vary their thermal treatments of ready-to-eat meat products in a safe manner. PMID- 12747690 TI - Reducing levels of Listeria monocytogenes contamination on raw salmon with acidified sodium chlorite. AB - The antimicrobial activity of acidified sodium chlorite (ASC) against Listeria monocytogenes in salmon was studied. Raw salmon (whole fish and fillets) inoculated with L. monocytogenes (10(3) CFU/cm2 or 10(4) CFU/g) were washed with ASC solution (50 ppm) for 1 min and stored at -18 degrees C for 1 month (whole salmon) or in ice for 7 days (fillets). L. monocytogenes populations were determined for whole salmon after frozen storage and for fillets on days 1, 3, 5, and 7 of storage. A wash with ASC solution followed by ASC glazing did not reduce L. monocytogenes on the skin of whole salmon during frozen storage. However, the wash resulted in an L. monocytogenes reduction of 0.5 log CFU/g for salmon fillets. The populations of L. monocytogenes in fillets increased slowly during ice storage, but the growth of these populations was retarded by ASC ice. By day 7, the populations were 0.25 log units smaller in fillets stored in ASC ice and 0.62 log units smaller in fillets that had been washed with ASC solution and stored in ASC ice than in control fillets. Treatment with ASC also reduced total plate counts (TPCs) by 0.43 log CFU/cm2 on the skin of whole salmon and by 0.31 log CFU/g in fillets. The TPCs for skin decreased during frozen storage but increased gradually for fillets stored at 5 degrees C or in ice. However, TPCs of ASC-treated samples were lower than those for controls at any point during the study. Washing with ASC solution significantly (P < 0.05) reduced TPCs on the skin of whole salmon and in fillets, as well as L. monocytogenes in fillets. The antimicrobial activity of ASC was enhanced when salmon was washed with ASC solution and stored in ASC ice. PMID- 12747691 TI - Gamma irradiation of fine-emulsion sausage containing sodium diacetate. AB - Listeria monocytogenes, a psychrotrophic foodborne pathogen, is a frequent postprocess contaminant of ready-to-eat (RTE) meat products, including frankfurters and bologna. Ionizing radiation can eliminate L. monocytogenes from RTE meats. Sodium diacetate (SDA) incorporated into fine-emulsion sausages inhibits the growth of L. monocytogenes. Irradiation of L. monocytogenes suspended in SDA solutions resulted in synergistic reductions of the microorganism. L. monocytogenes populations were reduced by > 9 log10 units at a radiation dose of 1.5 kGy when suspended in 0.125% SDA solution. In contrast, the D10-values (the ionizing radiation doses required to reduce the population by 90%) were 0.58, 0.59, 0.57, and 0.53 kGy for L. monocytogenes populations suspended in emulsions containing 0, 0.125, 0.25, and 0.5% SDA, respectively. The D10-values for L. monocytogenes surface inoculated onto frankfurters dipped in 0, 0.125, 0.25, and 0.5% SDA solutions were 0.58, 0.53, 0.54, and 0.52 kGy, respectively. Postirradiation growth of L. monocytogenes suspended in beef bologna emulsion at 9 degrees C was dependent on SDA concentration and ionizing radiation dose. Very small, but statistically significant, changes in bologna redness, lipid oxidation, and shear force were observed for the beef bologna emulsion with the highest SDA concentration (0.5%) and irradiation dose (3.0 kGy). SDA can inhibit the proliferation of L. monocytogenes surviving the irradiation process with minimal impact on fine-emulsion sausage color, lipid oxidation, and firmness when used within regulatory limits. PMID- 12747692 TI - Effect of dry heating on the microbiological quality, functional properties, and natural bacteriostatic ability of egg white after reconstitution. AB - Spray-dried egg white (powder) is widely used in the food industry because of its variety of functional properties and its practical advantages. Moreover, egg white powder is generally considered safe because it can withstand high temperatures that allow for the destruction of all pathogens, especially Salmonella. In France, two types of treatments are used to improve the functional properties (whipping and gelling) of dried egg white: standard storage at 67 degrees C for about 15 days and storage at 75 to 80 degrees C for 15 days. The objective of this study was to investigate the effects of two dry-heating treatments (storage at 67 and 75 degrees C for 15 days) on the subsequent ability of egg white to resist Salmonella growth after reconstitution. The impact on the endogenous microflora of the powder and on its functional properties was also considered. Both dry-heating treatments were efficient in destroying a large number of Salmonella. Dry heating at 75 degrees C affected the bacteriostatic ability of reconstituted egg white to a greater extent than did dry heating at 67 degrees C. This loss of bacteriostatic ability could be attributable to the thermal denaturation of ovotransferrin, resulting in a reduction in its activity as an iron chelator. However, dry heating at 75 degrees C resulted in improved functional properties. Ultimately, no complete compromise between better functional quality and the preservation of the bacteriostatic ability of egg white after reconstitution is possible. Our results underline the importance of the use of hygienic conditions with egg white powder, especially with powder subjected to high-temperature treatments. PMID- 12747693 TI - Growth and germination of proteolytic Clostridium botulinum in vegetable-based media. AB - The growth of proteolytic Clostridium botulinum from spore inocula and changes in spore counts in mushroom, broccoli, and potato purees were monitored. Four strains of proteolytic C. botulinum types A and B were inoculated separately at approximately 10(4) spores per ml in nutrient broth and vegetable purees incubated at 15, 20, and 30 degrees C for up to 52 days. The times for the cell populations to increase 1,000-fold (T1,000) in the tested vegetables (1 to 5 days at 30 degrees C, 3 to 16 days at 20 degrees C, 7 to > 52 days at 15 degrees C) were similar to those for meat or fish. Only temperature significantly influenced growth rate. In contrast, the lag phase depended on the strains and media tested, in addition to temperature. Lag times and T1,000S for proteolytic C. botulinum were longer for potato and broccoli purees than for mushroom puree. These differences were not related to different pHs or redox potentials. The germination level, evaluated as the decrease in the spore count, was low. The addition of a germinant mixture (L-cysteine, L-alanine, and sodium lactate) to some strains inoculated in vegetable purees resulted in an increase in germination, suggesting a lack of germination-triggering agents in the vegetable purees. PMID- 12747694 TI - Combined effect of cooking (grilling and roasting) and chilling storage (with and without air) on lipid and cholesterol oxidation in chicken breast. AB - The oxidation of the lipid fraction and cholesterol in raw and cooked chicken breast samples stored for 0 and 6 days at 4 degrees C under aerobic conditions and in vacuum packaging was studied. The multivariate statistical analysis showed significant effects of both culinary process and storage conditions on the lipid and cholesterol oxidation process, with a significant interaction between the two variables. Aerobic storage increased thiobarbituric acid reactive substances (TBA) from 0.04 to 0.06 ppm for raw samples, from 0.21 to 1.20 ppm for grilled samples, and from 0.24 to 1.62 ppm for roasted samples. During vacuum storage, only roasted samples showed significant increases in TBA. Levels of total cholesterol oxidation products (COP) remained low (2.88 to 4.35 microg/g of lipid) for all raw samples. Cooking increased COP levels to 12.85 and 11.54 microg/ g of lipid for grilled and roasted samples, respectively. Total COP and all individual COP except for cholestanetriol were significantly correlated with TBA and the peroxide index. However, the most extensive effect was attributable to the aerobic storage of cooked samples, which led to COP levels of 92.35 and 88.60 microg/g of lipid in grilled and roasted samples, respectively. Vacuum packaging did not increase COP levels for cooked samples. PMID- 12747695 TI - In vitro assessment of the cytotoxicity of nisin, pediocin, and selected colicins on simian virus 40-transfected human colon and Vero monkey kidney cells with trypan blue staining viability assays. AB - Gram-positive bacterial bacteriocins (nisin and pediocin) and gram-negative bacterial bacteriocins (colicins [Col] E1, E3, E6, E7, and K) were evaluated for cytotoxicity against cultured simian virus 40-transfected human colon (SV40-HC) and Vero monkey kidney (Vero) cells. Bacteriocin-treated cells were assessed for viability by trypan blue staining. Monolayers of SV40-HC and Vero cells were cultured in tissue culture plates (35 degrees C, 10% CO2 in humidified air) with the use of Dulbecco's modified Eagle's medium supplemented with 10% (vol/vol) calf serum. Actively growing cells in the log phase (ca. 10(4) cells per ml) were treated with individual partially purified bacteriocin preparations at 170, 350, and 700 activity units per ml. Duplicate culture plates for each bacteriocin treatment and untreated controls were withdrawn after 16, 32, and 48 h of incubation. Cells were dissociated with trypsin and treated with trypan blue and were then counted in a hemocytometer with the use of a phase-contrast microscope. Viability assays indicated dose-dependent toxicity for some bacteriocins. Nisin, pediocin, and Col E6 were the most cytotoxic bacteriocins; SV40-HC cells demonstrated greater sensitivity than Vero cells did. Some bacteriocins can be toxic to mammalian cells; therefore, bacteriocins intended for use as biopreservatives must be evaluated for toxicity to mammalian cells and for other toxicities. Col E1, Col E3, Col E7, and Col K demonstrated little toxicity at the activities tested, indicating that they are safe and thus have potential for use as food biopreservatives. PMID- 12747696 TI - Improving extraction of fumonisin mycotoxins from Brazilian corn-based infant foods. AB - The current AOAC International methods for the determination of fumonisins have been validated for corn and cornflakes but have produced low recoveries and high variability when applied to processed corn products for infants. Hence, an investigation was undertaken to improve the extraction efficiency for fumonisins by investigating the use of different extraction solvents. Corn-based infant foods containing cornmeal, corn starch, and corn flour were purchased in the city of Campinas, state of Sao Paulo, Brazil, and were analyzed for fumonisins B1 (FB1), B2 (FB2), and B3 (FB3) following extraction with a range of solvents. Comparison of the results from each of the samples indicated that acidified 70% aqueous methanol at pH 4.0 provided the best overall performance, whereas a methanol/boric acid (pH 9.2) mixture displayed poor extraction efficiency. Extraction with acidified 70% aqueous methanol showed seven of eight test samples to be positive for FB1 (range, 30 to 6,127 microg/kg; relative SD, 4.2 to 51.7%), two of eight samples to be positive for FB2 (range, 53 to 1,738 microg/kg; relative SD, 4.5 to 5.3%), and one of eight samples to be positive for FB3 (575 microg/kg). For samples in which extraction with phosphate-buffered mixtures (pH 3) proved superior, the method suffered from poor chromatography due to interfering compounds. The findings indicate that matrix interferences play a significant role in the extractability, cleanup, and chromatography of the fumonisins. PMID- 12747697 TI - Aflatoxin B1 and clinoptilolite in feed for laying hens: effects on egg quality, mycotoxin residues in livers, and hepatic mixed-function oxygenase activities. AB - Ninety-six laying hens were allocated to four groups administered different diets (group 0-0 received a complete diet, group 0-AF received a diet supplemented with 2.5 ppm of aflatoxin B1 [AFB1], group 2-0 received a diet supplemented with 2% clinoptilolite [CPL], and group 2-AF received a diet supplemented with 2% CPL and 2.5 ppm of AFB1) for 4 weeks to evaluate the effect of AFBI and/or CPL on egg quality and the ability of CPL to interact with the oral administration of AFB1. The possible effects of AFB1 on cytochrome P450-dependent hepatic mixed-function oxygenase (MFO) activities were also evaluated. Mycotoxin reduced yolk weight, while CPL influenced albumen percentage relative to that of eggs laid by chickens in group 0-AF Eggs laid by chickens in groups 0-AF and 2-AF had stronger shells and weighed less than the eggs of other groups. The eggs of treated groups were lighter in color than those of the control group (P < 0.01), and the tendency to yellowness in eggs was increased by CPL, probably through the affinity of red pigments for adsorbents and a consequent prevalence of yellow tonality. Color parameters might be connected with AFB1's interference with lipid metabolism and pigment deposition. The livers of hens in groups 0-AF and 2-AF showed very low mycotoxin concentrations that were significantly different (P < 0.01). The highest levels observed were those in the livers of the hens receiving the diet supplemented with the mycotoxin alone. AFB1 did not exert any significant effects on the hepatic MFO activities examined. PMID- 12747698 TI - Reduction of aflatoxins by Korean soybean paste and its effect on cytotoxicity and reproductive toxicity--Part 3. Inhibitory effects of Korean soybean paste (doen-jang) on aflatoxin toxicity in laying hens and aflatoxin accumulation in their eggs. AB - This study was conducted to determine the effects of Korean soybean paste (doen jang [dwen-jahng]) (at concentrations of 0.5, 1, and 5%) on the toxicity of 500 ppb of aflatoxin in the diets of 60 laying hens (Isa Brown) divided into five groups and treated from week 15 to week 67. The aflatoxin-treated hens exhibited many deleterious effects, including reduced body weight; increased relative organ weights; decreased egg production; aflatoxin accumulation in eggs; decreased serum calcium, phosphorus, and alanino amonotransferase (ALT) levels; increased serum gammaglutamil transferase and lactic dehydrogenase levels; and, most significantly, severely altered cell foci and sinusoid dilatation in the liver, relative to control hens. The feeding of 1% soybean paste to hens reduced the adverse effects of aflatoxin on body weight, relative organ weights, egg production, and aflatoxin accumulation in eggs and improved serum calcium and ALT levels and the histopathological lesions of the liver. The feeding of 5% soybean paste to hens resulted in higher levels of the same types of improvements, especially with regard to the histopathological findings for the liver. On the basis of these results, it was suggested that a diet including 5% (and in some cases only 1%) Korean soybean paste protected laying hens and their eggs from the major deleterious effects of 500 microg of aflatoxin per kg of diet and from aflatoxin accumulation. These results indicate that dietary supplementation with Korean soybean paste reduces aflatoxin toxicity in laying hens that ultimately produce human foods such as eggs and poultry. PMID- 12747699 TI - Impact of pH enhancement on populations of Salmonella, Listeria monocytogenes, and Escherichia coli O157:H7 in boneless lean beef trimmings. AB - Boneless lean beef trimmings were inoculated with multiple strains of salmonellae, Listeria monocytogenes, and Escherichia coli O157:H7 at levels of ca. 6 log10 CFU/g. pH enhancement with ammonia gas was then used to increase the pH of the trimmings to ca. 9.6. The product was then frozen, chipped, and compressed into blocks. pH enhancement reduced the populations of salmonellae, L. monocytogenes, and E. coli O157:H7 by approximately 4, 3, and 1 log10 cycles, respectively. After the product had been frozen and compressed into blocks, no salmonellae or E. coli O157:H7 were detectable by enumeration or after enrichment and isolation. The final populations of L. monocytogenes were reduced by ca. 3 log10 cycles relative to the initial populations. When uninoculated pH-enhanced lean boneless trimmings were blended with inoculated ground beef to a final concentration of 15% (wt/wt), pathogen populations in the ground beef were reduced by approximately 0.2 log10 cycles. PMID- 12747700 TI - Microbial quality of ostrich carcasses produced at an export-approved South African abattoir. AB - The aim of this study was to evaluate the microbial quality of ostrich carcasses produced in a South African export-approved ostrich abattoir. Ninety surface samples were collected from 30 ostrich carcasses at three processing points in the abattoir: after skinning, after evisceration, and after chilling. Samples were evaluated for aerobic plate counts, for levels of Pseudomonas spp., Enterobacteriaceae, and Staphylococcus aureus, and for the presence of Escherichia coli. Surface counts (means +/- standard deviations) at postskinning, postevisceration, and postchilling processing points were, respectively, 4.32 +/- 0.62, 4.21 +/- 0.63, and 4.57 +/- 0.48 log CFU/cm2 for total aerobes; 2.82 +/- 1.65, 2.86 +/- 1.53, and 3.75 +/- 0.94 log CFU/ cm2 for Pseudomonas spp.; 2.89 +/ 0.78, 2.90 +/- 0.53, and 2.38 +/- 0.67 log CFU/cm2 for S. aureus; and 2.55 +/- 1.53, 2.78 +/- 1.31, and 2.73 +/- 1.46 log CFU/cm2 for Enterobacteriaceae. Statistically significant differences were detected between the counts for the postskinning and postchilling processing points and between the counts for the postevisceration and postchilling processing points for total aerobes, Pseudomonas spp., and S. aureus. Of practical significance was the increase in Pseudomonas spp. counts on samples collected after chilling. Seventeen of 90 samples (18.8%) tested positive for E. coli. Counts for E. coli-positive samples ranged from 1.0 to 3.79 log CFU/cm2, with a mean count of 2.15 +/- 0.94 log CFU/cm2. The majority of the samples testing positive for E. coli were collected after evisceration. PMID- 12747701 TI - Aflatoxin contamination in shrimp feed and effects of aflatoxin addition to feed on shrimp production. AB - One hundred fifty samples of shrimp feed were collected from the eastern and southern regions of Thailand, and aflatoxins B1, B2, G1, and G2 (AFB1, AFB2, AFG1, and AFG2) in them were analyzed. AFB1 contamination ranged from a nondetectable level (< 0.003 ppb) to 0.651 ppb. Metabolites of AFB1 were less abundant than AFB1. To study the effects of aflatoxin in feed on shrimp production, black tiger shrimp were divided into four groups of 30 shrimp per group, tested in triplicate, and fed diets containing 0 (control), 5, 10, or 20 ppb of AFB1 for 10 consecutive days. After 7 or 10 days of consumption on each diet, the shrimp were weighed and sacrificed for laboratory examination. AFB1 and its metabolites were not detected in shrimp muscle. The mortality rate was slightly higher in the AFB1-treated groups than in the control group. The body weight of the surviving shrimp was decreased to 46 to 59% of the initial body weight in the AFB1-treated groups but not in the control group. Histopathological findings indicated hepatopancreatic damage by AFB1 with biochemical changes of the hemolymph. These results show that aflatoxin contamination in shrimp feed may cause economic losses by lowering the production of shrimp. Feed contaminated at the level of 20 ppb or lower (i.e., at the observed natural contamination level) may pose a very low risk, if any, to human health. PMID- 12747702 TI - Development of a specific monoclonal antibody for grouper (Epinephelus guaza) identification by an indirect enzyme-linked immunosorbent assay. AB - Identification of fish species adulteration is important for consumer protection and the enforcement of food-labeling laws. A monoclonal antibody (MAb) generated against soluble muscle proteins from grouper (Epinephelus guaza) has been used in two indirect enzyme-linked immunosorbent assay (ELISA) formats (microtiter plates and immunostick tubes) for the rapid authentication of grouper fillets. The 3D12 MAb was produced with the use of the hybridoma technique and tested against several commonly consumed fish species by ELISA. The 3D12 MAb specifically reacted with grouper samples and could be useful for the discrimination of grouper among other, less-valued, fish species sold in the marketplace. PMID- 12747703 TI - Autonomous nervous system with respect to dressing of cattle carcasses and its probable role in transfer of PrP(res) molecules. AB - Pathogen prions are widely recognized as the causative agent in bovine spongiform encephalopathy (BSE) and other transmissible spongiform encephalopathies. However, more research on the possible transmission mutes of this agent once it has reached the host is needed. There is evidence based on the anatomy and physiology of the autonomous nervous system (ANS), as well as observations for different animal species, that the ANS might be involved in the axonal drainage of pathogen prions toward the central nervous system. In this context, more attention should be paid to the cranial cervical ganglion, the stellate ganglion, the chain of paravertebral ganglia next to the first six thoracic vertebrae, the chain of the paravertebral ganglia next to loin vertebrae 1 through 6, the vagus nerve in the neck region and in the mediastine, and the esophagus (because of its close connection to the vagus nerve). For a more detailed risk analysis with respect to these tissues, the ANSs of animals having shown clinical signs of BSE might be examined to corroborate the evidence presented here. In the meantime, as a precautionary measure, the tissue addressed should be taken out of the human food chain, taken out of animal feed, and handled as if it were specified risk material. It is technically possible to remove these parts during cutting and dressing. PMID- 12747704 TI - In situ technique for measuring the orthogonality of a plane wave to a substrate. AB - A new compact in situ method of measuring the perpendicularity of a plane wave to a substrate is proposed. Off-axis cylindrical Fresnel lenses are used to focus a portion of the incident plane wave onto target lines. The displacement of the focal line from the targets is determined by the degree of angular misalignment. The proposed design has been incorporated into a 10-mm-thick fused-silica module, which enables us to obtain an alignment precision of better than 0.0083 degrees. This method is designed for use in optical assembly procedures that require an incident collimated beam that is normal to the alignment features. Experimental results are presented. PMID- 12747705 TI - Eigenmodes of fractional Hankel transform derived by the entangled-state method. AB - Using the entangled-state method in quantum mechanics, we find that the eigenmodes of the fractional Hankel transform are two-variable Hermite-Gaussian functions that can be rewritten in a clearer form as Laguerre polynominals. PMID- 12747707 TI - Theory and practice of long-period gratings: when a loss becomes a gain. AB - Losses of cladding modes are part of the mechanism of operation of a long-period grating (LPG) when it is used as an optical filter. We present a LPG computer simulation that accounts for these losses. On the basis of this simulation, we show that losses result in qualitatively different LPG spectral behavior. There is an optimal loss value that provides sidelobe-free, 100% power transfer from the core to the cladding mode for a uniform LPG. We obtained a simple equation that relates this optimum lose value to the LPG length and the cross-coupling coefficient. Based on the results, we propose new approaches to LPG design in a fiber as well as in waveguide platforms for fiber-optic communication and sensor applications. A design of a LPG reconfigurable filter is suggested. PMID- 12747706 TI - Measurement of phase differences between the diffracted orders of deep relief gratings. AB - Measurement of the phase difference between the 0th and the 1st transmitted diffraction orders of a symmetrical surface-relief grating recorded on a photoresist film is carried out by replacement of the grating in the same setup with which it was recorded. The measurement does not depend on lateral shifts of thereplaced grating relative to the interference pattern, on environmental phase perturbations or on the wave-front quality of the interfering beams. The experimental data agree rather well with theoretical results calculated for sinusoidal profiled gratings. PMID- 12747708 TI - Scattering at sidewall roughness in photonic crystal slabs. AB - We have simulated the effect of sidewall roughness in photonic-crystallike structures with different vertical refractive-index contrast. We treated the scattering off a sidewall irregularity as a radiating dipole excited by the incident waveguide mode. We show that the loss that is due to this scattering is significantly larger for structures with a low refractive-index contrast (such as GaAs/AlGaAs waveguides) than for structures with a high vertical index contrast (such as silicon-on-insulators and membranes). PMID- 12747709 TI - Continuous-wave fiber optical parametric wavelength converter with +40-dB conversion efficiency and a 3.8-dB noise figure. AB - We have obtained 40 dB of internal (on-off) conversion gain and a sub-4-dB noise figure (NF) with a continuous-wave (cw) fiber optical parametric wavelength converter. To our knowledge, this is the lowest NF reported for any cw wavelength converter. We have also investigated the properties of NF versus signal input power and pump power. PMID- 12747711 TI - Optical sectioning in wide-field microscopy obtained by dynamic structured light illumination and detection based on a smart pixel detector array. AB - Optical sectioning in wide-field microscopy is achieved by illumination of the object with a continuously moving single-spatial-frequency pattern and detecting the image with a smart pixel detector array. This detector performs an on-chip electronic signal processing that extracts the optically sectioned image. The optically sectioned image is directly observed in real time without any additional postprocessing. PMID- 12747710 TI - Confocal fluorescence spectroscopy and anisotropy imaging system. AB - We report the design and implementation of a laser scanning confocal fluorescence system with spectroscopy and anisotropy imaging capabilities. Confocal spectroscopy is achieved with a fiber pinhole that is inserted into and removed from the detection path as needed. Fluorescence anisotropy imaging is accomplished with a polarizing beam splitter placed after the conventional pinhole. Two orthogonal polarizations are detected simultaneously with balanced photomultiplier tubes. The quality of the axial sectioning that is achieved in the confocal fluorescence spectroscopy mode is demonstrated experimentally, and examples of polarization-sensitive fluorescence imaging are demonstrated in tumor cell monolayers. PMID- 12747712 TI - Projected index computed tomography. AB - Projected index computed tomography (PICT) is a new imaging technique that provides a computed reconstruction of the index of refraction of a sample. PICT makes use of data from standard optical coherence tomography images taken from several view angles to determine a mapping of the refractive indices of the sample. A rectilinear propagation model is assumed, so the data are understood to be related to the line integral of the refractive index in the beam paths. These data thus provide a set of angular projections of the sample. The spatial distribution of the index of the object may then be reconstructed by use of standard filtered backprojection techniques. The resultant PICT images are free of the spatial distortion that is inherent in standard optical cross-sectional images and correspond well to the manufactured dimensions of specific samples. PMID- 12747713 TI - Free-running 9.1-microm distributed-feedback quantum cascade laser linewidth measurement by heterodyning with a C18O2 laser. AB - We report spectral linewidth measurements of a 9.1-microm distributed-feedback quantum cascade laser (QCL). The free-running QCL beam was mixed with a waveguide isotopic C18O2 laser onto a high-speed HgCdTe photomixer, and beat notes were recorded from a radio-frequency spectral analyzer. Beating was performed at two operating conditions, first near the QCL laser threshold (beating with the C18O2 R10 line) and then at a high injection current (beating with the C18O2 R8 line). Overall, beat note widths of 1.3-6.5 MHz were observed, which proves that a free running QCL can have a short-term spectral width near 1 MHz. PMID- 12747714 TI - Compact, broad-bandwidth fiber laser for sub-2-microm axial resolution optical coherence tomography in the 1300-nm wavelength region. AB - A novel, compact, user friendly fiber laser with a broad emission bandwidth (MenloSystems, lambdac = 1375 nm, deltalambda = 470 nm, Pout = 4 mW) was used to achieve unprecedented sub-2-microm axial resolution optical coherence tomography (OCT) in nontransparent biological tissue in the 1300-nm wavelength region. Fresh human skin and arterial biopsies were imaged ex vivo with approximately 1.4 microm axial and approximately 3-microm lateral resolution and 95-dB sensitivity, demonstrating the great potential for clinical OCT applications of this stable, low-cost, and turn-on-key fiber laser. PMID- 12747715 TI - Spatial solitons in optically induced gratings. AB - We study experimentally nonlinear localization effects in optically induced gratings created by interfering plane waves in a photorefractive crystal. We demonstrate the generation of spatial bright solitons similar to those observed in arrays of coupled optical waveguides. We also create pairs of out-of-phase solitons, which resemble twisted localized states in nonlinear lattices. PMID- 12747716 TI - Local second-harmonic generation enhancement on gold nanostructures probed by two photon microscopy. AB - The localization of surface second-harmonic generation (S-SHG) enhancements from granular gold structures that exhibit local plasmon resonance was investigated. A two-photon microscopy technique was used to perform high spatial resolution S-SHG imaging. The magnitude and the spatial density of S-SHG enhancement confined in submicroscopic regions are strongly dependent on the morphology of the gold's surface. Polarization measurements of local S-SHG responses reveal the local field anisotropy in enhancement regions and furthermore prove the incoherent and strongly depolarized nature of the emission, which is attributed to ultrafast fluctuations of the enhancement location in the focal volume. PMID- 12747717 TI - Second-harmonic generation from ellipsoidal silver nanoparticles embedded in silica glass. AB - Second-harmonic generation of uniformly oriented, ellipsoidal silver nanoparticles in a glass matrix was observed and investigated as a function of incidence angle, light polarization, and spatial arrangement of the particles. The results can be explained by the symmetry of the spatial nanoparticle arrangement and by resonance enhancement that is due to the localized surface plasmons of the particles. Second-harmonic enhancement is observed only in sufficiently thin layers (deltakl < pi); on a sample with two separate layers, strong modulation owing to quasi-phase matching is obtained. PMID- 12747718 TI - Ultrashort 1.5-microm laser excited upconverted stimulated emission based on simultaneous three-photon absorption. AB - Simultaneous three-photon excited stimulated emission at (cavityless lasing) approximately 610 nm has been achieved in an organic chromophore solution pumped by approximately 1.5-microm ultrashort coherent radiation. The stimulated emission can be observed only in the forward and backward directions and is characterized by its high directionality and spectral narrowing. The divergence angle for the backward stimulated emission is considerably smaller than that for the forward stimulated emission, which resembles optical phase conjugation. PMID- 12747719 TI - Comparison of the solutions from a novel variational method with numerical results for the study of beam propagation in a Kerr medium with nonlinear absorption. AB - We present results from a novel variational method for the study of beam propagation in a Kerr medium with nonlinear absorption. This new method combines the variational method and a nonlinear absorption equation and gives a concise expression for the combination. The results obtained with this method show good quantitative agreement with numerical solutions obtained with the finite difference method. It is shown that the variational method takes much less time than a numerical simulation with the finite-difference method for analysis of beam propagation in a thick medium with nonlinear absorption and nonlinear refraction. The new method makes detailed analysis of beam propagation in a Kerr medium with nonlinear absorption very simple and fast. PMID- 12747720 TI - Determination of small anisotropy of holographic phase gratings. AB - We show the possibility of detecting small anisotropies in holographic polymer dispersed liquid-crystal samples, using a simple experimental setup that allows us to determine the behavior of the diffraction efficiency versus incident angle for two reading polarizations. This analysis is extremely sensitive to small changes in the parameters that define the grating anisotropy, giving us a way to determine with great accuracy the components of the modulated part of the dielectric tensor. PMID- 12747721 TI - Substrate-strain-induced tunability of dense wavelength-division multiplexing thin-film filters. AB - Classic dense wavelength-division multiplexing thin-film filters can be spectrally tuned through the substrate's strain. We analyze the theoretical shift of the design wavelength of a narrow-bandpass filter when uniform, uniaxial compressive stress is applied to the substrate, and we compare calculated sensitivity with experimental data. We measure the transmittance shape of a 200 GHz standard filter for several loading cases to quantify the increase of insertion losses. PMID- 12747722 TI - Simple reflow technique for fabrication of a microlens array in solgel glass. AB - A simple reflow method for fabrication of refractive microlens arrays in inorganic-organic SiO2-ZrO2 solgel glass is presented. To our knowledge, this is the first report that presents a simple reflow technique for transforming a negatively induced hybrid solgel material into desirable spherical microlenses. It is shown that the microlenses have excellent smooth surfaces and uniform dimensions. The reflow technique is considerably cheaper than use of a high energy beam-sensitive gray-scale mask and is suitable for mass production. PMID- 12747723 TI - Reduction of propagation and bending losses of heterostructured photonic crystal waveguides by use of a high-delta structure. AB - We describe the fabrication and evaluation of a low-loss, high-delta optical waveguide consisting of heterostructured photonic crystals. The waveguide is composed of a multilayer stack of Ta2O5/SiO2 and is prepared by use of the autocloning technique. Light is guided in the waveguide by the difference in the effective refractive indices of the constituent photonic crystals. By improving the design of the core region so that is has a flat multilayer structure, we achieve delta = 3.09% for the in-plane direction and net propagation loss of 0.56 dB/mm at lambda = 1.6 microm. Experiments also suggest that the bending loss of the waveguide can be reduced to less than 0.1 dB if the curvature radius is larger than 700 microm. PMID- 12747724 TI - Optical doughnut for optical tweezers. AB - We describe novel optical doughnuts for optical tweezers. With new phase functions, the proposed doughnut beams have dark cores in specified shapes. The technique can offer a simple method for creating a variety of beam shapes to match the trapped objects. One can rotate the beams directly by revolving their phase structures about their axes on the initial plane. The technique for generating the traditional Laguerre-Gaussian beam can be used to create these novel beams. PMID- 12747725 TI - Coherent measurement of short laser pulses based on spectral interferometry resolved in time. AB - We propose a new method for measuring the phase and the amplitude of a short laser pulse that is based on shearing interferometry in the spectral domain combined with time gating. The method has several features in common with spectral interferometry for direct electric-field reconstruction, in particular, fast and direct reconstruction of the phase. Accurate measurement of the phase added to an 80-fs pulse by a block of F4 glass demonstrates the technique. PMID- 12747726 TI - Control of relative carrier-envelope phase slip in femtosecond Ti:sapphire and Cr:forsterite lasers. AB - We were able to control relative carrier-envelope phase slip among mode-locked Ti:sapphire and Cr:forsterite lasers by employing electronic feedback. The pulse timings of these lasers were passively synchronized with our crossing-beam technique. Since the optical-frequency ratio of Ti:sapphire and Cr:forsterite is approximately 3:2, we can observe the phase relation by superimposing the third harmonic of Cr:forsterite and the second harmonic of Ti:sapphire lasers in time and in space. The spectrum width of the locked beat note was less than 3 kHz, which corresponds to the controlled fluctuation of a cavity-length difference of less than 10 pm. PMID- 12747727 TI - Tunable optimal compression of ultrabroadband pulses by cross-phase modulation. AB - We show how cross-phase modulation between two pulses, combined with optimal pulse shaping at the input of a dielectric medium, can be used to generate nearly single-cycle pulses that are tunable from the ultraviolet to the mid-infrared at the output of the medium, precompensating for dispersion to all orders. PMID- 12747728 TI - 2nd International Meeting on Free Radicals in Health and Disease. The role of oxidants and antioxidants in the regulation of chronic diseases, may 8-12, 2002, Istanbul, Turkey. PMID- 12747729 TI - Derivatives of xanthic acid are novel antioxidants: application to synaptosomes. AB - Xanthic acids have long been known to act as reducing agents. Recently, D609, a tricyclodecanol derivative of xanthic acid, has been reported to have anti apoptotic and anti-inflammatory properties that are attributed to specific inhibition of phosphatidyl choline phospholipase C (PC-PLC). However, because oxidative stress is involved in both of these cellular responses, the possibility that xanthates may act as antioxidants was investigated in the current study. Finding that xanthates efficiently scavenge hydroxyl radicals, the mechanism by which D609 and other xanthate derivatives may protect against oxidative damage was further examined. The xanthates studied, especially D609, mimic glutathione (GSH). Xanthates scavenge hydroxyl radicals and hydrogen peroxide, form disulfide bonds (dixanthogens), and react with electrophilic products of lipid oxidation (acrolein) in a manner similar to GSH. Further, upon disulfide formation, dixanthogens are reduced by glutathione reductase to a redox active xanthate. Supporting its role as an antioxidant, D609 significantly (p < 0.01) reduces free radical-induced changes in synaptosomal lipid peroxidation (TBARs), protein oxidation (protein carbonyls), and protein conformation. Thus, in addition to inhibitory effects on PC-PLC, D609 may prevent cellular apoptotic and inflammatory cascades by acting as antioxidants and novel GSH mimics. These results are discussed with reference to potential therapeutic application of D609 in oxidative stress conditions. PMID- 12747731 TI - Targeting superoxide dismutase to renal proximal tubule cells attenuates vancomycin-induced nephrotoxicity in rats. AB - Vancomycin hydrochloride (VCM), a glycopeptide antibiotic, has a broad spectrum against methicillin-resistant Staphylococcus aureus (MRSA). As it is known to induce renal dysfunction, the dose and the duration of its administration are limited. Moreover, the mechanism of VCM-induced renal dysfunction remains to be unclear. To evaluate the involvement of free radical on VCM-induced renal dysfunction, we carried out analysis with a hexamethylenediamine-conjugated superoxide dismutase (AH-SOD) which rapidly accumulates in renal proximal tubule cells and inhibits oxidative injury of the kidney. Male Wistar rats (weighing 200 210 g) were intraperitonealy administered with 200 mg/kg of VCM twice a day for 7 days. AH-SOD 5 mg/kg/day was subcutaneously injected 5 min before every VCM injection. VCM induced renal injury dose-dependently. Biochemical analyses revealed that plasma levels of blood urea nitrogen and creatinine significantly increased in the VCM-treated group by an AH-SOD-inhibitable mechanism. VCM simultaneously elicited an increase of 8-OHdG levels and chemiluminescence intensity of free radical generation in the kidney. Histological examination revealed that VCM also elicited a marked destruction of glomeruli and necrosis of proximal tubules. AH-SOD inhibited these phenomena in the kidney. These results suggested that oxidative stress might underlie the pathogenesis of VCM-induced nephrotoxicity and targeting SOD and/or related antioxidants to renal proximal tubules might permit the administration of higher doses of VCM sufficient for eradication of MRSA without causing renal injury. PMID- 12747730 TI - Oxidative damage of rat cerebral cortex and hippocampus, and changes in antioxidative defense systems caused by hyperoxia. AB - In order to elucidate the oxidative damage in rat brain caused by oxidative stress, regional changes in the levels of lipid peroxidation products and antioxidative defense systems in cerebral cortex and hippocampus, and in their synapses, which modulate learning and memory functions in the brain, were studied. When rats were subjected to hyperoxia as an oxidative stress, thiobarbituric acid reactive substance (TBARS) in the regions studied increased more than in normal rats by approximately 35%. The values in oxygen-unexposed vitamin E-deficient rats were also higher than in normal rats. It was found that the TBARS contents in synaptosomes isolated from both regions were remarkably higher than in the organs. These results imply that synapses are more susceptible to oxidative stress than the organ itself. This tendency was also observed in the content of conjugated diene. In response to oxidative stress, the status of the antioxidant defense system in each region, i.e. the concentration of vitamin E, and the activities of superoxide dismutase, catalase and glutathione peroxidase, decreased remarkably. On the other hand, in oxygen-unexposed vitamin E-deficient rats, the activities of these enzymes each region tended to increase, except for catalase activity. These results suggest that in response to the oxidative stress, the antioxidant defense systems may be consumed to prevent oxidative damage, and then, may be supplied through the antioxidant network. PMID- 12747732 TI - Angeli's salt induces neurotoxicity in dopaminergic neurons in vivo and in vitro. AB - In this study, we investigated the hypothesis that the pro-oxidative properties of Angeli's salt (AS), a nitroxyl anion (HNO/NO-) releasing compound, cause neurotoxicity in dopaminergic neurons. The pro-oxidative properties were demonstrated in vitro by measuring hydroxylation products of salicylate and peroxidation of lipids under various redox conditions. AS (0-1000 microM) released high amounts of hydroxylating species in a concentration dependent manner. AS also increased lipid peroxidation in brain homogenates at concentrations below 100 microM, while inhibiting it at 1000 microM concentration. The AS induced pro-oxidative effects were completely suppressed by copper (II), which converts nitroxyl anion to nitric oxide, as well as by a potent nitroxyl anion scavenger glutathione. Neurotoxicity towards dopaminergic neurons was tested in rat nigrostriatal dopaminergic system in vivo and by using primary mesencephalic dopaminergic neuronal cultures in vitro. Intranigral infusion of AS (0-400 nmol) caused neurotoxicity reflected as a dose dependent decrease of striatal dopamine seven days after treatment. The effect of the 100 nmol dose was more pronounced whenmeasured 50 days after the infusion. Neurotoxicity was also confirmed as a decrease of tyrosine hydroxylase positive neurons in the substantia nigra. Neither sulphononoate, a close structural analog of AS, nor sodiumnitrite caused changes in striatal dopamine, thus reflecting lack of neurotoxicity. In primary dopaminergic neuronal cultures AS reduced [3H] dopamine uptake with concentrations over 200 microM confirming neurotoxicity. In line with the quite low efficacy to increase lipid peroxidation in vitro, infusion of AS into substantia nigra did not cause increased formation of fluorescent products of lipid peroxidation. These results support the hypothesis that AS derived species oxidize critical thiol groups, rather than membrane lipids, potentially leading to protein oxidation/dysfunction and demonstrated neurotoxicity These findings may have pathophysiological relevance in case of excess formation of nitroxyl anion. PMID- 12747733 TI - Irradiation of cells with ultraviolet-A (320-400 nm) in the presence of cell culture medium elicits biological effects due to extracellular generation of hydrogen peroxide. AB - Biological effects of ultraviolet A (UVA) irradiation have been ascribed to the photochemical generation of singlet oxygen. Not all effects described in the literature, however, are explicable solely by the generation of singlet oxygen, but rather resemble effects elicited by hydrogen peroxide (H2O2). Here, we show that when cells are kept in cell culture media during exposure to UVA, stress kinases, including ERK 1 and ERK 2 as well as Akt (protein kinase B), are activated, whereas there is no or only minor activation when cells are kept in phosphate-buffered saline during irradiation. Indeed, the exposure of cell culture media to UVA (30 J/cm2) results in the generation of significant amounts of H2O2, with concentrations of about 100 microM. H2O2 concentrations are at least three-fold higher in HEPES-buffered culture media after UVA irradiation. From experiments with solutions of riboflavin, tryptophan or HEPES, as well as combinations thereof, it is concluded that riboflavin mediates the photooxidation of either tryptophan or HEPES, resulting in the generation of H2O2. Thus, if signaling effects of UVA radiation are to be investigated in cell culture systems, riboflavin and HEPES/tryptophan should be avoided during irradiation because of artificial H2O2 generation. It should be taken into account, however, that in vivo tryptophan and riboflavin might play an important role in the generation of reactive oxygen species by UVA as both substances are abundant in living tissues. PMID- 12747734 TI - Antioxidant activity of galloyl quinic derivatives isolated from P. lentiscus leaves. AB - The antioxidant properties of galloyl quinic derivatives isolated from Pistacia lentiscus L. leaves have been investigated by means of Electron Paramagnetic Resonance spectroscopy (EPR) and UV-Vis spectrophotometry. Antioxidant properties have been also estimated using the biologically relevant LDL test. The scavenger activities of gallic acid, 5-O-galloyl, 3,5-O-digalloyl, 3,4,5-O-trigalloyl quinic acid derivatives, have been estimated against 1,1-diphenyl-2 picrylhydrazyl (DPPH) radical, superoxide (O2) radical, and hydroxyl (OH) radical. On the whole, the scavenger activity raised as the number of galloyl groups on the quinic acid skeleton increased. The half-inhibition concentrations (IC50) of di- and tri-galloyl derivatives did not exceed 30 microM for all the tested free radicals. All the tested metabolites strongly reduced the oxidation of low-density lipoproteins (LDL), following a trend similar to that observed for the scavenger ability against OH radical. PMID- 12747735 TI - Membrane oxidation assay--a novel lipoxygenase-based evaluation of membrane oxidizability. AB - A membrane oxidation assay is presented which uses isolated erythrocyte membranes ("ghosts") alipoxygenase as a selective catalyst for the transfer of oxygen to cis-cis-1,4-pentadiene-moieties. The latter are, for instance, present in linoleic and arachidonic acids, both of which are integral parts of membranes. These non-conjugated double bonds represent energetically favorable sites for oxidative attack and therefore, may be rearranged and partially consumed during oxidative stress. Consequently, the measurement of oxygen consumption in the course of the lipoxygenase-mediated oxidation provides a tool for the quick and reliable determination of such double bonds. Significant inter-individual differences have been noted in 11 subjects, which also correlate to the total radical antioxidant parameter (TRAP) values obtained. This assay will be helpful in the assessment of oxidizable structures in erythrocyte membranes that may be diminished as a consequence of oxidative damage suffered by an individual. In conclusion, a simple and rapid assay for the assessment of the oxidizability of erythrocyte membranes is presented complementing the TRAP assay for plasma antioxidative status. PMID- 12747736 TI - Effect of docosahexaenoic acid and ascorbate on peroxidation of retinal membranes of ODS rats. AB - Mutant male osteogenic disorder Shionogi (ODS) rats, unable to synthesize ascorbic acid, were fed diets containing a high content of docosahexaenoic acid (DHA) and different amounts of ascorbic acid, to study the effect of DHA on peroxidative susceptibility of the retina and possible antioxidant action of ascorbic acid. ODS rats were fed from 7 weeks of age with diets containing high DHA (6.4% of total energy). A control group received a diet high in linoleic acid. The diets also contained varying amounts of ascorbic acid. Fatty acid compositions and phospholipid hydroperoxides in rod outer segment (ROS) membranes, and retinal ascorbic acid were analyzed. DHA in ROS membranes was significantly increased in rats fed high DHA, compared with the linoleic acid diet. Levels of phospholipid hydroperoxides in the DHA-fed rats were significantly higher than the linoleic acid-fed rats. Ascorbic acid supplementation did not suppress the phospholipid hydroperoxide levels after a high DHA diet, even when the supplement increased the content of retinal ascorbic acid. In conclusion, high DHA feeding induced a marked increase of phospholipid hydroperoxides in ROS membranes of ODS rats. Supplementation of ascorbic acid did not reverse this increase. PMID- 12747738 TI - Alpha tocopherol protects against immunosuppressive and immunotoxic effects of lead. AB - Chronic exposure to lead (Pb) is associated with multiorgan toxicity. The precise mechanism(s) involved, however, remains incompletely defined. The present study was undertaken to analyze the effect of Pb on the immune system and determine the ability of alpha tocopherol (AT) to reverse Pb-induced immunotoxicity. Groups of TO Mice (6 per group) were treated ip for 2 weeks with saline alone, Pb acetate alone, Pb plus AT, or with AT alone. Spleens were then analyzed for (i) cellular composition by flow cytometry, (ii) cellular response to B and T cell mitogens and (iii) production of nitric oxide (NO). Pb treatment resulted in a significant state of splenomegaly associated mainly with an influx of CD11b+ myeloid cells. Surprisingly, however, these cells exhibited no upregulation in expression of activation markers and did not produce NO. The lymphocyte mitogenic responses were inhibited by > or = 70% in Pb-treated group. Concurrent treatment with Pb and AT resulted in almost a complete reversal of Pb-induced splenic cellular influx. Despite this, however, mitogenic responses in Pb + AT treated group were approximately 50% of those observed in normal (saline-treated) controls. We conclude that (1) chronic treatment with Pb acetate induces a state of splenomegaly and decreased proliferation in response to mitogenic stimuli and (2) co-treatment with AT largely reversed the cellular influx but this was associated with only a partial improvement of the mitogenic responses. These results highlight the role of AT as a potentially effective antioxidant in the immune system. PMID- 12747737 TI - Lipid peroxidation inhibition reduces NF-kappaB activation and attenuates cerulein-induced pancreatitis. AB - Increased lipid peroxidation, enhanced nuclear factor kappa-B (NF-kappaB) activation and augmented tumor necrosis factor-alpha (TNF-alpha) production have been implicated in cerulein-induced pancreatitis. We investigated whether lipid peroxidation inhibition might reduce NF-kappaB activation and the inflammatory response in cerulein-induced pancreatitis. Male Sprague-Dawley rats of 230-250g body weight received administration of cerulein (80 microg/kg s.c. for each of four injections at hourly intervals). A control group received four s.c. injections of 0.9% saline at hourly intervals. Animals were randomized to receive either raxofelast, an inhibitor of lipid peroxidation (20 mg/kg i.p. administered with the first cerulein injection) or its vehicle (1 ml/kg of a 10% DMSO/NaCl solution). All these rats were sacrificed 2 h after the last injection of either cerulein or its vehicle. Raxofelast administration (20 mg/kg i.p. with the first cerulein) significantly reduced malondialdehyde (MDA) levels, an index of lipid peroxidation (CER + DMSO = 3.075 +/- 0.54 micromol/g; CER + raxofelast = 0.693 +/ 0.18 micromol/g; p < 0.001), decreased myeloperoxidase (MPO) activity (CER + DMSO = 22.2 +/- 3.54 mU/g; CER + raxofelast = 9.07 +/- 2.05 mU/g, p < 0.01), increased glutathione levels (GSH) (CER + DMSO = 5.21 +/- 1.79 micromol/g; CER + raxofelast = 15.71 +/- 2.14 micronol/g; p < 0.001), and reduced acinar cell damage evaluated by means of histology and serum levels of both amylase (CER + DMSO = 4063 +/- 707.9 U/l; CER + raxofelast = 1198 +/- 214.4 U/l; p < 0.001), and lipase (CER + DMSO = 1654 +/- 330 U/l; CER + raxofelast = 386 +/- 118.2 U/l; p < 0.001), Furthermore, raxofelast reduced pancreatic NF-kappaB activation and the TNF-alpha mRNA levels and tissue content of mature protein in the pancreas. Indeed, lipid peroxidation inhibition might be considered a potential therapeutic approach to prevent the severe damage in acute pancreatitis. PMID- 12747739 TI - Free-radical scavengers and antioxidants from Peumus boldus Mol. ("Boldo"). AB - The dry leaves of Peumus boldus (Monimiaceae) are used in infusion or decoction as a digestive and to improve hepatic complains. Preliminary assays showed free radical scavenging activity in hot water extracts of boldo leaves, measured by the decoloration of a methanolic solution of the 1,1-diphenyl-2-picryl-hydrazyl radical (DPPH). Assay-guided isolation led to the active compounds. Catechin proved to be the main free-radical scavenger of the extracts. Lipid peroxidation in erythrocytes was inhibited by boldo extracts and fractions at 500 microg/ml with higher effect for the ethyl acetate soluble and alkaloid fractions. The IC50 for catechin and boldine in the lipid peroxidation test were 75.6 and 12.5 microg/ml, respectively. On the basis of dry starting material the catechin content in the crude drug was 2.25% while the total alkaloid calculated as boldine was 0.06%. The activity of boldine was six times higher than catechin in the lipid peroxidation assay. However, the mean catechin:total alkaloid content ratio was 37:1. The relative concentration of alkaloids and phenolics in boldo leaves and their activity suggest that free-radical scavenging effect is mainly due to catechin and flavonoids and that antioxidant effect is mainly related with the catechin content The high catechin content of boldo leaves and its bioactivity suggest that quality control of Boldo folium has to combine the analysis of catechin as well as their characteristic aporphine alkaloids. PMID- 12747740 TI - Cyanidin-3-O-beta-glucopyranoside protects myocardium and erythrocytes from oxygen radical-mediated damages. AB - The cyanidin-3-O-beta-glucopyranoside (C-3-G) antioxidant capacity towards reactive oxygen species (ROS)-mediated damages was assessed in tissue and cells submitted to increased oxidative stress. In the isolated ischemic and reperfused rat heart, 10 or 30 degreesM C-3-G protected from both lipid peroxidation (66.7 and 94% inhibition of malondialdehyde (MDA) generation in 10 and 30 microM C-3-G reperfused hearts, respectively, in comparison with control reperfused hearts) and energy metabolism impairment (higher ATP concentration in 10 and 30 microM C 3-G-reperfused hearts than in control reperfused hearts). These effects were associated to C-3-G permeation within myocardial cells, as indicated by results obtained in the isolated rat heart perfused for 30 min in the recirculating Langendorff mode under normoxia with 10 and 30 microM C-3-G. Protective effects were exerted, in a dose-dependent manner, by C-3-G also in 2 mM hydrogen peroxide treated human erythrocytes. With respect to MDA formation, an apparent IC50 of 5.12 microM was calculated for C-3-G (the polyphenol resveratrol used for comparison showed an apparent IC50 of 38.43 microM). The general indications are that C-3-G (largely diffused in dietary plants and fruits, such as pigmented oranges very common in the Mediterranean diet) represents a powerful natural antioxidant with beneficial effects in case of increased oxidative stress, and at pharmacological concentrations it is able to decrease tissue damages occurring in myocardial ischemia and reperfusion. PMID- 12747741 TI - Pulmonary bioavailability of ascorbic acid in an ascorbate-synthesising species, the horse. AB - Vitamin C (ascorbic acid) is a non-enzymatic antioxidant important in protecting the lung against oxidative damage and is decreased in lung lining fluid of horses with airway inflammation. To examine possible therapeutic regimens in a species with ascorbate-synthesising capacity, we studied the effects of oral supplementation of two forms of ascorbic acid, (each equivalent to 20 mg ascorbic acid per kg body weight) on the pulmonary and systemic antioxidant status of six healthy ponies in a 3 x 3 Latin square design. Two weeks supplementation with ascorbyl palmitate significantly increased mean plasma ascorbic acid concentrations compared to control (29 +/- 5 and 18 +/- 7 micromol/l, respectively; p < 0.05). Calcium ascorbyl-2-monophosphate, a more stable form of ascorbic acid, also increased mean plasma ascorbic acid concentrations, but not significantly (23 +/- 1 micromol/l; p = 0.07). The concentration of ascorbic acid in bronchoalveolar lavage fluid increased in five out of six ponies following supplementation with either ascorbyl palmitate or calcium ascorbyl-2 monophosphate compared with control (30 +/- 10, 25 +/- 4 and 18 +/- 8 micromol/l, respectively; p < 0.01). Neither supplement altered the concentration of glutathione, uric acid or alpha-tocopherol in plasma or bronchoalveolar lavage fluid. In conclusion, the concentration of lung lining fluid ascorbic acid is increased following ascorbic acid supplementation (20 mg/kg body weight) in an ascorbate-synthesising species. PMID- 12747742 TI - Cancer Vaccine Collaborative 2002: opening address. PMID- 12747743 TI - Cell surface expression of heat shock protein gp96 enhances cross-presentation of cellular antigens and the generation of tumor-specific T cell memory. AB - Gp96 is an endoplasmic reticular heat shock protein (HSP). We have shown previously that surface expression of gp96 (96tm) on tumor cells led to the activation of dendritic cells and increased anti-tumor immunity. In this report, we have found that protective immunity elicited by 96tm+ tumor cells was tumor specific and long-lasting. Both CD4+ and CD8+ T cell memory were elicited. By immunizing with tumor cells loaded with the chicken ovalbumin (ova) model antigen, we demonstrated that the priming of adoptively transferred ova-specific CD8+ T cells could occur across MHC haplotypes. The efficiency of this cross priming can be significantly increased when mice were immunized with whole cells that express both ova and cell surface gp96 (ova+96tm+). Mere mixture of soluble ova with 96tm-expressing tumor cells (ova-96tm+) was insufficient, arguing for further processing of ova and perhaps the participation of 96tm-ova complexes in this process. We further compared the relative efficiency of two whole cell vaccines based on the manipulation of gp96 expression in one system: 96tm+ whole cells and cells that secrete the gp96-Ig fusion protein. We found that both vaccines are effective in a prophylactic model against tumors. Our study has reinforced the notion that the manipulation of the site of expression of HSPs may be an effective approach for cancer immunotherapy. PMID- 12747744 TI - Concordant down-regulation of proto-oncogene PML and major histocompatibility antigen HLA class I expression in high-grade prostate cancer. AB - Recognition of tumor cells by cytolytic T lymphocytes depends on cell surface MHC class I expression. As a mechanism to evade T cell recognition, many malignant cancer cells, including those of prostate cancer, down-regulate MHC class I. For the majority of human cancers, the molecular mechanism of MHC class I down regulation is unclear, although it is well established that MHC class I down regulation is often associated with the down-regulation of multiple genes devoted to antigen presentation. Since the promyelocytic leukemia (PML) proto-oncogene controls multiple antigen-presentation genes in some murine cancer cells, we analyzed the expression of proto-oncogene PML and MHC class I in high-grade prostate cancer. We found that 30 of 37 (81%) prostate adenocarcinoma cases with a Gleason grade of 7-8 had more than 50% down-regulation of HLA class I expression. Among these, 22 cases (73.3%) had no detectable PML protein, while 4 cases (13.3%) showed partial PML down-regulation. In contrast, all 7 cases of prostate cancer with high expression of cell surface HLA class I had high levels of PML expression. Concordant down-regulation of HLA and PML was observed in different histological patterns of prostate adenocarcinoma. These results suggest that in high-grade prostate cancer, malfunction of proto-oncogene PML is a major factor in the down-regulation of cell surface HLA class I molecules, the target molecules essential for the direct recognition of cancer cells by cytolytic T lymphocytes. PMID- 12747745 TI - MUC1-like tandem repeat proteins are broadly immunogenic in cancer patients. AB - The identification of antigens mediating tumor rejection is an important goal of cancer immunology. The SEREX technology utilizes antibodies from cancer patients to identify candidate antigens from tumor-derived cDNA expression libraries. Using sera from a long-term surviving metastatic melanoma patient vaccinated with irradiated, autologous tumor cells engineered to secrete granulocyte-macrophage colony stimulating factor (GM-CSF), we identified an antigen reported to be a putative opioid growth factor receptor (OGFr). The human immune response to OGFr exhibits three features shared with other tumor antigens. First, the protein is an intracellular antigen found in both nucleus and cytoplasm. Second, part of the antibody response is directed at a putative protein product encoded by an alternative reading frame (ARF). Third, part of the antibody response is directed at a portion of the molecule that bears a striking resemblance to the extracellular domain of MUC1, both with respect to primary structure and size polymorphism. Antibody responses to OGFr and a synthetic peptide representing a putative alternative reading frame product (OGFr-ARF) were frequently found in cancer patients. 11/45 (24%) melanoma patients had antibodies to OGFr and 5/45 (11%) had antibodies to OGFr-ARF. Moreover, 5/24 (21%) lung cancer, 4/25 (16%) prostate cancer, and 5/6 breast or ovarian cancer patients had antibodies to OGFr, the alternative frame product, or both. These data add to the growing list of tumor antigens that appear to be translated in two frames, and suggest that OGFr and OGFr-ARF may be useful targets for vaccination. PMID- 12747746 TI - Depletion of CD25+ regulatory cells results in suppression of melanoma growth and induction of autoreactivity in mice. AB - Treatment with monoclonal antibodies (mAbs) specific for CD25 (anti-CD25 mAb) has been shown to suppress growth of a variety of different tumours in mice. These studies did not however determine whether or not anti-CD25 mAbs facilitate tumour rejection by depletion of regulatory T cells or by binding to tumour-specific effector cells. Using a murine model of melanoma we have found that treatment of mice with anti-CD25 mAb facilitates long-term CD4+ T cell-mediated tumour immunity through depletion of CD25+ regulatory cells. We further show that the effector CD4+ T cells confer long-term tumour immunity even in the presence of CD25+ regulatory cells and do not require CD8+ T cells for tumour rejection. The inhibitory impact of anti-CD25 mAb treatment on tumour growth may be the result of depleting CD25+ regulatory cells that normally inhibit the generation of immune responses to self-antigens that are shared by the tumour. We have performed experiments to determine whether or not immune responses to melanocyte antigens are generated in anti-CD25 mAb-treated, melanoma-immune mice. The results of the experiments indicate that a T cell response to the melanocyte antigen tyrosinase accompanies suppression of tumour growth in mice lacking CD25+ regulatory cells. PMID- 12747747 TI - CD40 triggering increases the efficiency of dendritic cells for antitumoral immunization. AB - Recent studies have shown the importance of triggering CD40 molecules to enhance the efficiency of dendritic cells (DCs) as antigen-presenting cells (APCs). The P198 and P1A tumor antigens, which are expressed by mastocytoma P815, have been assessed for their immunogenicity using different modes of immunization. We measured CTL responses induced in vivo with antigenic peptides P198 and P1A loaded onto bone marrow-derived DCs that had matured as a consequence of CD40 CD40L interactions. CD40L-transfected 3T3 fibroblasts were used as a source of CD40L signal. Our results show that this mode of DC activation considerably improves their ability to induce CTLs against P198 and P1A antigens in vivo as compared to untreated DCs. We also show that immunizations carried out with CD40L activated DCs loaded with the P1A peptide induce a very efficient protection against a lethal challenge with P815 tumor cells, which express P1A. Our results indicate that the efficiency of DC-based vaccines used in clinical trials of cancer immunotherapy could be increased significantly by triggering DCs via CD40 prior to immunization. PMID- 12747749 TI - Immunization against a dominant tumor antigen abrogates immunogenicity of the tumor. AB - To study the role of subdominant epitopes in tumor rejection we have used EL4 tumor cells and their ovalbumin (OVA)-transfected counterpart E.G7. Immunization of mice with irradiated EL4 cells conferred protection against challenge with EL4 and E.G7. Surprisingly, immunization with irradiated E.G7 cells did not protect against a subsequent challenge with EL4 or E.G7. Growth of E.G7 tumors in E.G7 immunized mice was not due to loss of expression of OVA or MHC I by the tumor cells in vivo. Adoptive transfer of OVA-specific transgenic T cells, immunization of mice with native or heat-denatured OVA or infection with a recombinant virus expressing OVA also failed to induce rejection of E.G7 tumors. Lack of immunogenicity of the OVA-expressing tumor could not be overcome by combination of a CD40 activating antibody with immunization against E.G7 or OVA. Our results suggest that immunization against subdominant epitopes is more effective than vaccination against dominant epitopes. PMID- 12747748 TI - In vitro generated cytolytic T lymphocytes reactive against head and neck cancer recognize multiple epitopes presented by HLA-A2, including peptides derived from the p53 and MDM-2 proteins. AB - In previous studies, we were successful in generating HLA-A2-restricted CD8+ CTLs reactive with head and neck carcinomas (HNCs) in 4/10 cases using traditional mixed lymphocyte tumor cultures (MLTCs) employing a semi-allogeneic HLA-A2+ HNC cell line, PCI-13, as the stimulator of normal HLA-A2+ donor T lymphocytes. However, these T cell lines contained only 1-1.5% HLA-A2-restricted, tumor reactive CD8+ CTLs, as assessed by both limiting dilution and IFN-gamma ELISPOT assays. In order to increase the success rate in generating such HNC-reactive CTL lines, we modified the procedure to allow for T cell crosspriming by autologous DCs pulsed with PCI-13 lysates. In all three attempts, HLA-A2-restricted effector T cell lines were obtained that contained PCI-13-reactive CD8+ T cells at frequencies as high as 1 in 6. These cultured bulk lines recognized at least five predominant HLA-A2-restricted epitopes based on ELISPOT fingerprinting of HPLC fractionated, naturally presented PCI-13-derived peptides. Two of these epitopes appear to be derived from the p53 and MDM-2 proteins overexpressed by the PCI-13 cell line. Interestingly, the synthetic wild type sequence p53 (264-272) and MDM 2 (53-61) peptides were able to drive in vitro generation of tumor-specific CTLs from the PBMCs of normal HLA-A2+ donors. However, this MDM-2 peptide was not able to elicit responses from HLA-A2+ patients with HNC in short-term in vitro cultures. Overall, these data suggest that tumor lysate-loaded DCs elicit a broad repertoire of CTL responses, some of which are directed against peptides derived from cell cycle regulatory proteins that may prove to be of clinical significance in the therapy of HNC. PMID- 12747750 TI - Identification of tumor-restricted antigens NY-BR-1, SCP-1, and a new cancer/testis-like antigen NW-BR-3 by serological screening of a testicular library with breast cancer serum. AB - Serological analysis of recombinant cDNA expression libraries (SEREX) has led to the identification of several categories of new tumor antigens. We analyzed a testicular cDNA expression library with serum obtained from a breast cancer patient and isolated 13 genes designated NW-BR-1 through NW-BR-13. Of these, 3 showed tumor-restricted expression (NW-BR-1, -2 and -3), the others being expressed ubiquitously. NW-BR-3, representing 9 of 24 primary clones, showed tissue-restricted mRNA expression, being expressed in normal testis but not in 15 other normal tissues tested by Northern blotting. RT-PCR analysis showed strong NW-BR-3 expression in normal testis, weak expression in brain, kidney, trachea, uterus and normal prostate, and was negative in liver, heart, lung, colon, small intestine, bone marrow, breast, thymus, muscle, spleen, and stomach. NW-BR-3 mRNA expression was found in different tumor tissues and tumor cell lines by RT-PCR, thus showing a 'cancer/testis' (CT)-like mRNA expression pattern. NW-BR-3 shares 71% nucleotide and amino acid homology to a mouse gene cloned from mouse testicular tissue. Based on the mRNA expression pattern, NW-BR-3 represents a new candidate target gene for cancer immunotherapy. NW-BR-1 and NW-BR-2 also showed tumor-restricted mRNA expression. NW-BR-1 is a partial clone of the breast differentiation antigen NY-BR-1 previously identified by SEREX. NY-BR-1 is expressed in normal breast, testis and 80% of breast cancers. NW-BR-2 is identical to the CT antigen SCP-1, initially isolated by SEREX analysis of renal cancer. This study provides further evidence that SEREX is a powerful tool to identify new tumor antigens potentially relevant for immunotherapy approaches. PMID- 12747751 TI - T-cell response to unique and shared antigens and vaccination of cancer patients. AB - Most vaccination studies of cancer patients find no clear association between clinical and immunological responses to the vaccine. We discuss the possible kinetics of the T cell response in melanoma patients against unique or shared tumor antigens. We hypothesize that a response against unique antigens prevails during primary melanoma growth, causing the selection of tumor cells lacking most of these antigens unless these are necessary to maintain the neoplastic state. After a subset of tumor cells metastasize to the lymph nodes, T cells are activated against previously ignored shared, differentiation-like antigens, owing to a new environment where pro-inflammatory cytokines can be present. The development of a T cell response to such normal epitopes then associates with tumor growth, but remains clinically inefficient. We predict that two immunologically different subsets of melanoma patients may exist, one that mounted an early immune response against melanoma antigens and one that did not. A paradox may emerge when vaccination is attempted in these two groups of subjects, with the second group being more prone to develop an effective immune response if the vaccine is potent enough to activate naive T cells, while the first has probably already eliminated most of the tumor antigens potentially recognizable by the host T cells owing to the previous selection made by the immune response developed early during tumor growth. Thus, it is likely that the subgroup of metastatic patients with a high frequency of anti-melanoma memory T cells may not show a clinical response to vaccination. PMID- 12747752 TI - Functional analysis of HLA-A*0201/Melan-A peptide multimer+ CD8+ T cells isolated from an HLA-A*0201- donor: exploring tumor antigen allorestricted recognition. AB - Recent studies in mouse models have suggested that genetic transfer of tumor antigen-specific high affinity T cell receptors (TCR) into host lymphocytes could be a viable strategy for the rapid induction of tumor-specific immunity. A previously proposed approach for the isolation of such TCRs consists in circumventing tolerance to self-restricting HLA/peptide complexes by deriving them from PMBCs of allogenic donors. Towards this aim, we used fluorescent HLA-A2 class-I/peptide soluble multimers to isolate A2-restricted CD8+ T cells specific for a previously described Melan-A peptide enhanced analog (Melan-A 26-35 A27L) from an HLA-A*0201 (A2) negative donor. We isolated two distinct groups of Melan A 26-35 A27L-specific clones. Clones from the first group recognized the analog peptide with high avidity but showed very low recognition of Melan-A parental peptides. In contrast, clones from the second group efficiently recognized Melan A parental peptides. Surprisingly however, most clones recognized not only A2+ Melan-A+ targets, but also A2+ Melan-A- targets suggesting that they can also recognize endogenous peptides other than Melan-A. In addition, one clone showed full cross-recognition of an antigenically unrelated peptide. Together, our data show that HLA-A2/peptide multimers can be successfully used for the isolation of allorestricted CD8+ T cells reactive with tumor antigen-derived peptides. However, as the cross-reactivity of these apparently peptide-specific allorestricted TCRs is presently unpredictable, a careful in vitro analysis of their reactivity to the host's normal cells is recommended. PMID- 12747753 TI - Induction of autoantibodies against tyrosinase-related proteins following DNA vaccination: unexpected reactivity to a protein paralogue. AB - DNA vaccination against tissue-restricted antigens is a strategy for cancer therapy. Immune tolerance and ignorance of self antigens has been a hurdle for this approach. We have shown that immunization with xenogeneic DNA orthologues elicits tumor immunity. One model that we have developed entails immunization of mice against tyrosinase-related protein-2 (Tyrp2) using cDNA encoding homologous human Tyrp2. A subset of mice immunized with human Tyrp2 developed antibody responses to Tyrp1. Unexpectedly, this was not simply due to cross-reactivity, as mice with anti-Tyrp1 antibodies were not usually the same animals with anti-Tyrp2 antibodies. Although autoimmune vitiligo was frequently observed in mice that had been immunized with Tyrp2, its occurrence was not correlated with the development of antibodies to Tyrp1. This implies that the appearance of anti-Tyrp1 antibodies was not simply a consequence of the destruction of melanocytes by T-cells recognizing Tyrp2. This represents an example of intermolecular determinant recognition, but is not simply due to epitope spreading since antibodies against the antigen targeted by DNA vaccination are not typically detected. PMID- 12747754 TI - Identification of a new peptide recognized by autologous cytolytic T lymphocytes on a human melanoma. AB - Melanoma line LG2-MEL expresses several antigens recognized by autologous CTLs. One of them consists of a peptide derived from tyrosinase and presented by HLA B*3503. We have identified another antigen of LG2-MEL as a peptide presented by HLA-B*4403 and resulting from a point mutation in gene OS-9. This gene is expressed in various normal tissues. It is located on chromosome 12 in the vicinity of the CDK4 locus and is frequently co-amplified with CDK4 in human sarcomas. The mutation, a C-to-T transition, changes a proline residue into a leucine at position 446 of the OS-9 protein. Mutated transcripts were found in all the melanoma sublines of LG2-MEL. None of the 184 tumor samples collected from other cancer patients expressed the mutated transcript, indicating that this is a rare mutational event. Interestingly, some of the melanoma sublines of LG2 MEL have lost the wild-type allele of gene OS-9. Those sublines appear to grow faster in vitro than the sublines that retained the wild-type allele, suggesting that this loss of heterozygosity may favor tumor progression. The mutation we have identified in gene OS-9 might therefore participate in the oncogenic process by affecting the function of this potential tumor-suppressor gene. PMID- 12747755 TI - Urine antibody against human cancer antigen NY-ESO-1. AB - NY-ESO-1 is one of the most immunogenic tumor antigens known to date. Spontaneous humoral and cellular immune responses against NY-ESO-1 are detected in a substantial proportion of patients with NY-ESO-1 positive cancers. NY-ESO-1 serum antibody is dependent on the presence of NY-ESO-1+ cancer cells, and antibody titers correlate with the clinical development of the disease. NY-ESO-1 serum antibody is associated with detectable NY-ESO-1-specific CD8+ T cell reactivity. High titers of NY-ESO-1 serum antibodies are found in patients with advanced NY ESO-1+ malignancies. Urine samples of seropositive patients with normal kidney function were tested for NY-ESO-1 antibody by Western blotting and enzyme-linked immunosorbent assay (ELISA). Antibodies to NY-ESO-1 were found in the urine of patients whose NY-ESO-1 serum antibody titers were 1:10,000 or higher by Western blotting. In patients with weak (positive at 1:250, negative at 1:1,000) or no reactivity, urine antibody was not detectable. No urine NY-ESO-1 antibody was found in patients without detectable NY-ESO-1 serum antibody. Our results show that urine analysis for NY-ESO-1 antibody identifies patients with strong NY-ESO 1 immunity. Urine antibody detection may also be of value in the monitoring of spontaneous and vaccine-induced immunity against other defined tumor antigens. PMID- 12747756 TI - Expression of cancer-testis genes in human hepatocellular carcinomas. AB - Cancer-testis (CT) genes are expressed in a variety of human cancers, but not in normal tissues except for testis, and represent promising targets for immunotherapy and gene therapy. We investigated the expression of 10 CT genes (MAGE-1, MAGE-3, MAGE-4, GAGE, NY-ESO-1, SSX-1, HOM-MEL-40/SSX-2, SSX-4, HOM-TES 14/SCP-1, and HOM-TES-85) in 21 hepatocellular carcinoma (HCC) biopsy specimens. The most frequently expressed CT genes were SSX-1 and GAGE, which were found in 8/21 (38%) HCC samples, followed by HOM-TES-14/SCP-1 (6/21 or 29%), MAGE-3 (5/21 or 24%), HOM-TES-85 and MAGE-1 (4/21 or 19% each), whereas SSX-4 and HOM-MEL 40/SSX-2 were only expressed in 2/21 cases each, MAGE-4 in one case, and NY-ESO-1 not at all. Of the 21 HCC cases investigated, only four did not express any of the CT genes tested, 17 (81%) expressed at least one, 9 (43%) coexpressed two, four (19%) coexpressed four, three (14%) coexpressed five and one coexpressed 8 of the 10 CT genes tested. We conclude that a majority of HCC cases might be amenable to specific immunotherapeutic interventions. However, the identification of additional tumor-specific antigens with a frequent expression in HCCs is warranted to develop widely applicable, polyvalent HCC vaccines. PMID- 12747757 TI - Identification of a naturally processed NY-ESO-1 peptide recognized by CD8+ T cells in the context of HLA-B51. AB - NY-ESO-1 is one of the most immunogenic cancer antigens known to date, inducing humoral and cellular immune responses in a high proportion of patients with advanced NY-ESO-1-expressing cancers. The assessment of spontaneous and vaccine induced CD8+ T cell responses has been limited to a small number of known NY-ESO 1 epitopes presented by MHC class I alleles. Recently, a new method to monitor NY ESO-1-specific CD8+ T cell responses was introduced that does not depend on the individual MHC class I status and on predefined peptide epitopes. Antigen presenting cells transduced with recombinant adenoviral vectors encoding NY-ESO-1 were used to stimulate CD8+ selected NY-ESO-1-specific T cells. Effector cells were tested for recognition of autologous B cell targets transfected with NY-ESO 1 using a recombinant vaccinia virus construct. Using a modified approach we identified the NY-ESO-1 p94-102 peptide as being recognized by CD8+ T cells in the context of HLA- B51. NY-ESO-1 p94-102 specific CD8+ T cells recognized naturally processed NY-ESO-1 presented by HLA-B51+ monocyte-derived dendritic and tumor cells. Transfection of target cells with NY-ESO-1 combined with different HLA class I alleles confirmed that the NY-ESO-1 peptide was naturally processed and recognized by HLA-B51-restricted CD8+ T cell lines and clones. Therefore, NY ESO-1 p94-102 is a new candidate peptide antigen for cancer immunotherapy and for the monitoring of spontaneous and vaccine-induced NY-ESO-1-specific T cell responses in HLA- B51+ patients with NY-ESO-1 expressing malignancies. PMID- 12747758 TI - Cytokine enhancement of in vitro antibody-dependent cellular cytotoxicity mediated by chimeric anti-GD3 monoclonal antibody KM871. AB - The chimeric KM871 monoclonal antibody targets the GD3 disialoganglioside antigen and is under investigation for its immunotherapeutic potential in melanoma. Preclinical and phase I studies have demonstrated the biodistribution and specific tumour targeting of KM871 to metastatic melanoma in vivo, with a long half-life and lack of immunogenicity making it an attractive candidate for further clinical trials. In vitro studies have demonstrated KM871 induces high levels of cytotoxicity in both antibody-dependent cellular-cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) assays. In order to investigate the potential for cytokine upregulation of KM871-mediated ADCC, freshly isolated healthy donor PBMC effector cells were cultured in the presence or absence of the cytokines interleukin-2, interleukin-12 and granulocyte/macrophage-colony stimulating factor and the ADCC determined over a 10-day period. In the absence of these cytokines, ADCC activity of 1 micro g/ml KM871 on (51) Cr-labeled SK-MEL 28 target cells could not be detected after 72 hrs of culture of PBMC effector cells in media. In contrast, ADCC mediated by KM871 was significantly enhanced and maintained for the 10-day study period upon culturing PBMCs with media containing IL-2 and/or IL-12, but not with GM-CSF. FACS analysis of the effector cell population indicated CD3-/CD16+56+ NK cells were primarily responsible for the KM871-mediated ADCC activity and a direct correlation was observed between the percentage of NK cells and the level of cytotoxicity mediated by the PBMCs. Furthermore, ADCC was significantly reduced using NK-depleted PBMCs. These results suggest combining IL-2 or IL-12 with KM871 may enhance KM871 immune mediated cell killing in patients with metastatic melanoma. PMID- 12747759 TI - Can hTERT peptide (540-548) -specific CD8 T cells recognize and kill tumor cells? AB - This commentary reviews the data on HLA-A2-restricted CD8 T cells specific for peptide (540-548) derived from hTERT (human telomerase reverse transcriptase). Several studies have reported the successful generation of such T cells (1, 2, 3). However, tumor recognition was observed in some, but not all, studies. More data are required to elucidate whether hTERT peptide (540-548) -specific T cells can indeed recognize and destroy tumor cells. It would be highly useful if telomerase would emerge as a universal tumor antigen that can be targeted in the cancer immunotherapy of HLA-A2 positive patients. PMID- 12747760 TI - Characterization of cells prepared by dendritic cell-tumor cell fusion. AB - Dendritic cells (DCs) are professional antigen-presenting cells currently being discussed as a potent tool for antitumor vaccination strategies. The approach consisting of the in vitro generation of DC-tumor cell hybrids may be advantageous for individualized vaccines since there is no need for the determination of MHC-restricted tumor-associated antigens recognized by T cells. As recent vaccination studies gave varying results, we tested the impact of the fusion treatment on the cells used. Polyethylene glycol-induced fusion, as well as electrofusion, proved to be suitable for generating hybrid cells although at a low frequency. Of note, both methods also gave rise to DCs having phagocytosed apoptotic tumor cells. The expression of surface molecules relevant for specific T cell stimulation was not altered by the fusion procedure and the DCs were still functionally active as demonstrated by the secretion of IL-12 and the uptake of antigen. The cells were able to induce a tumor-specific T cell response in vitro and therefore deserve further investigation as potent tools for immunotherapy trials. PMID- 12747761 TI - Naturally formed or artificially reconstituted non-covalent alpha2-macroglobulin peptide complexes elicit CD91-dependent cellular immunity. AB - Immunization of mice with in vitro reconstituted alpha2-macroglobulin-peptide complexes primes peptide-specific CTL responses. We show here using the H-Y antigenic system that naturally produced, immunogenic alpha2-macroglobulin peptide complexes can be isolated from the sera of normal male mice. As an application of these ideas to cancer immunity, we show that the immunity evoked by alpha2-macroglobulin-peptide complexes reconstituted in vitro is effective in prophylaxis against tumors. Furthermore, complex peptide mixtures isolated from tumor lysates can be reconstituted non-covalently with alpha2-macroglobulin and such complexes elicit potent protective tumor immunity. This approach circumvents the need for prior knowledge of the identity of the immunogenic peptides. The heat shock protein/alpha2-macroglobulin receptor CD91 is shown to be involved in the ability of heat shock proteins or alpha2-macroglobulin to elicit an anti tumor immune response. PMID- 12747762 TI - Cancer/testis (CT) antigens - a new link between gametogenesis and cancer. PMID- 12747763 TI - Redirecting anti-viral CTL against cancer cells by surface targeting of monomeric MHC class I-viral peptide conjugated to antibody fragments. AB - To combine the advantage of both the tumor targeting capacity of high affinity monoclonal antibodies (mAbs) and the potent killing properties of cytotoxic T lymphocytes (CTL), we investigated the activity of conjugates made by coupling single Fab' fragments, from mAbs specific for tumor cell surface antigens, to monomeric HLA-A2 complexes containing the immunodominant influenza-matrix peptide 58-66. In solution, the monovalent 95 kDa Fab-HLA-A2/Flu conjugates did not activate influenza-specific CTL. However, when targeted to tumor cells expressing the relevant tumor-associated antigen, the conjugates induced CTL activation and efficient tumor cell lysis, as a result of MHC/peptide surface oligomerization. The highly specific and sensitive in vitro cytotoxicity results presented suggest that injection of Fab-MHC/peptide conjugates could represent a new form of immunotherapy, bridging antibody and T lymphocyte attack on cancer cells. PMID- 12747764 TI - Towards a cancer immunome database. PMID- 12747765 TI - Humoral immunity to human breast cancer: antigen definition and quantitative analysis of mRNA expression. AB - The ability of the immune system to recognize structurally altered, amplified or aberrantly expressed proteins can be used to identify molecules of etiologic relevance to cancer and to define targets for cancer immunotherapy. In the current study, ninety-four distinct antigens reactive with serum IgG from breast cancer patients were identified by immunoscreening breast cancer-derived cDNA expression libraries (SEREX). A serological profile was generated for each antigen on the basis of reactivity with allogeneic sera from normal individuals and cancer patients, and mRNA expression profiles for coding sequences were assembled based upon the tissue distribution of expressed sequence tags, Northern blots and real-time RT-PCR. Forty antigens reacted exclusively with sera from cancer patients. These included well-characterized tumor antigens, e.g. MAGE-3, MAGE-6, NY-ESO-1, Her2neu and p53, as well as newly-defined breast cancer antigens, e.g. kinesin 2, TATA element modulatory factor 1, tumor protein D52 and MAGE D, and novel gene products, e.g. NY-BR-62, NY-BR-75, NY-BR-85, and NY-BR-96. With regard to expression profiles, two of the novel gene products, NY-BR-62 and NY-BR-85, were characterized by a high level of testicular mRNA expression, and were overexpressed in 60% and 90% of breast cancers, respectively. In addition, mRNA encoding tumor protein D52 was overexpressed in 60% of breast cancer specimens, while transcripts encoding SNT-1 signal adaptor protein were downregulated in 70% of these cases. This study adds to the growing list of breast cancer antigens defined by SEREX and to the ultimate objective of identifying the complete repertoire of immunogenic gene products in human cancer (the cancer immunome). PMID- 12747766 TI - Identification of a tumor-associated contact-dependent activity which reversibly downregulates cytolytic function of CD8+ T cells. AB - Tumors elicit an immune response in hosts and yet, paradoxically, often grow progressively with fatal consequences. This phenomenon has been attributed to the possible expression by tumor cells of immunomodulatory factors that overcome the anti-tumor effector functions of both specific and non-specific immune cells. This study reports on the ability of the methylcholanthrene-induced fibrosarcoma, Meth A, as well as other tumors of varied histological origins to downregulate the lytic activity of CD8+ T cells. The suppressive activity is contact-dependent and reversible. As tumor-bearing hosts are rarely immunosuppressed systemically, these findings may explain how local events within the tumor bed subvert the specific anti-tumor immune response. PMID- 12747767 TI - The strange road to the tumor-specific transplantation antigens (TSTAs). PMID- 12747768 TI - Determinants of efficacy of immunotherapy with tumor-derived heat shock protein gp96. AB - Immunotherapy with gp96 was highly effective in mice bearing methylcholanthrene induced fibrosarcomas (Meth A tumors) when treatment began 7 days or less after tumor challenge, but significantly less effective if the treatment began 9 days after challenge. Immunotherapy of pre-existing tumors showed all the hallmarks of specificity of gp96 and dose-restriction observed previously with prophylactic studies. When mice with large primary Meth A tumors were treated with surgery alone, or with surgery followed by therapy with Meth A-derived gp96, the mice that received surgery and immunotherapy did significantly better than those receiving surgery alone. The relationship between the time of initiation of immunotherapy with gp96 and its efficacy was also tested in a metastatic model of the Lewis lung carcinoma. In this model, immunotherapy with gp96 was very effective if treatment began up to 31 days after tumor challenge, but significantly less so if therapy was initiated day 33 post-tumor challenge. These observations suggest that the regulatory phenomena that interfere with immunotherapy gather momentum with surprising speed. PMID- 12747769 TI - ELISPOT cloning of tumor antigens recognized by cytotoxic T-lymphocytes from a cDNA expression library. AB - The methodology of cloning genes coding for antigens recognized by T-cells from cDNA expression libraries was improved technically by using enzyme-linked immunospot (ELISPOT) assays instead of enzyme-linked immunosorbent assays (ELISA) or bioassays to detect cytokines produced by T-cells in response to antigens. Combining large and small scale ELISPOT assays for expression cloning has the following advantages compared to conventional cDNA expression cloning: i) the number of recombinant plasmids which can be screened is greater than 10,000 per well in a 24-well plate in a large scale ELISPOT assay compared to fewer than 100 per well in a 96-well plate in an IFN-gamma ELISA or a TNF-alpha bioassay; ii) the total number of recombinant plasmids which can be screened in a routine assay is 2 x 10 (5) in only one 24-well plate in a large scale ELISPOT assay compared to 1 x 10 (5) in ten 96-well plates in an IFN-gamma ELISA or a TNF-alpha bioassay. Thus the screening efficiency of large scale ELISPOT cloning is approximately 200 times that of conventional expression cloning approaches. The efficiency of the method was confirmed by detecting the model gene RLakt from a cDNA library of a murine leukemia RL male 1. PMID- 12747770 TI - Alternative roles for interferon-gamma in the immune response to DNA vaccines encoding related melanosomal antigens. AB - Tyrosinase-related proteins-1 and -2 (gp75/TRP-1 and TRP-2) are melanosomal membrane glycoproteins recognized by antibodies and T-cells from patients with melanoma. Xenogeneic DNA immunization against gp75/TRP-1 generates antibody dependent tumor immunity and autoimmune depigmentation. In contrast xenogeneic TRP-2 DNA immunization induces immunity mediated by CD8+ T-cells. The role of IFN gamma in the generation of tumor immunity and autoimmune depigmentation in these two models was investigated. No tumor protection and minimal depigmentation was observed after immunization with human TRP-2 DNA in mice deficient in IFN-gamma ligand. Repletion with recombinant murine IFN-gamma restored tumor immunity. Experiments using IL4 deficient mice demonstrated that tumor immunity was unaffected but that autoimmune depigmentation was potentially accelerated, consistent with down-modulation of autoimmunity against TRP-2 by IL4. In contrast, IFN-gamma was not required for the generation of immunity to gp75/TRP 1. In fact, exogenous IFN-gamma ablated autoantibody responses against gp75/TRP-1 after xenogeneic DNA immunization, consistent with a down-regulatory effect of IFN-gamma. These results show that immunity to TRP-2 following DNA immunization uses an IFN-gamma-dependent Th1 pathway, but immunity to gp75/TRP-1 is down regulated by IFN-gamma. PMID- 12747772 TI - Systematic search and molecular characterization of the antigenic targets of myeloma immunoglobulins: a monoclonal IgA from a female patient targeting sperm specific cylicin II. AB - The identification of the antigenic stimuli of B-cell neoplasms might be of considerable importance since a causal relationship between these neoplasms and antigenic stimulation has been suggested. To date the identification of such antigens has been erratic and accidental. For a systematic search and molecular characterization of human proteins that are antigenic target structures of myeloma-associated immunoglobulins, we applied SEREX (serological analysis of antigens by recombinant cDNA expression cloning) using a testis cDNA expression library and myeloma proteins from 42 patients. A monoclonal IgA from a female patient was shown to target sperm-specific cylicin II. The specificity of the reaction was confirmed by the characteristic staining of the equatorial belt of human sperm heads by the patient's myeloma protein. Serological analysis of recombinantly expressed cDNAs is a straightforward and high throughput approach for the molecular characterization of the targets of myeloma-associated immunoglobulins. The analysis of the antigenic spectrum of immunoglobulins associated with B-cell neoplasms will provide valuable information for the understanding of the pathogenesis of these diseases. PMID- 12747771 TI - Minimal costimulatory requirements for T cell priming and TH1 differentiation: activation of naive human T lymphocytes by tumor cells armed with bifunctional antibody constructs. AB - Direct priming of naive human CD8+ and CD4+ T cells by tumor cells devoid of any intrinsic antigen presentation properties, but passively armed with recombinant proteins mediating primary and costimulatory T cell signals, was investigated. Bifunctional antibody constructs were used to specifically target costimulatory molecules such as B7-1, B7-2 and LFA-3 to the epithelial cell adhesion molecule (EpCAM), a surface antigen successfully used as target for antibody therapy of minimal residual colorectal cancer. T cell priming was monitored by flow cytometric analysis of CD45 isoform expression and confirmed by measuring typical effector functions of primed T cells known to be absent from naive T lymphocytes. Accordingly, CD8+ T cells were tested for cytotoxic activity and secretion of TNF alpha, while secretion of IFN-gamma, IL-5 and IL-4 was determined for CD4+ T cells. B7, known to be required for the initial activation of naive T cells, also proved to be sufficient for T cell priming when present as the only costimulatory molecule together with an appropriate primary signal. The requirement of dendritic and other antigen presenting cells (APCs) for T cell priming through non-APCs such as tumor cells could be ruled out. Under minimal priming conditions, naive CD4+ T cells were found to exclusively enter the TH1 developmental pathway, while several factors thought to favor TH2 polarization, like weak primary signals and B7-2 versus B7-1 costimulation, could be excluded as dominant TH2 promoters. PMID- 12747773 TI - Understanding the structure-activity relationship of the human ether-a-go-go related gene cardiac K+ channel. A model for bad behavior. PMID- 12747774 TI - Identification of 1-arylmethyl-3- (2-aminoethyl)-5-aryluracil as novel gonadotropin-releasing hormone receptor antagonists. AB - Based on SAR from bicyclic GnRH antagonists such as 6-aminomethyl-7 arylpyrrolo[1,2-a]pyrimid-4-ones (1) and 2-aryl-3-aminomethylimidazolo[1,2 a]pyrimid-5-ones (2a,b), a series of novel uracil compounds (4) were derived as the GnRH antagonists. Their syntheses and initial SAR are discussed herein. This is the first time that monocycle-based GnRH receptor antagonists are reported. PMID- 12747775 TI - Synthesis, structure-activity relationship, and biological studies of indolocarbazoles as potent cyclin D1-CDK4 inhibitors. AB - Novel substituted indolocarbazoles were synthesized, and their kinase inhibitory capability was evaluated in vitro. 6-Substituted indolocarbazoles 4 were found to be potent and selective D1/CDK4 inhibitors. 4d and 4h exhibited potent and ATP competitive D1/CDK4 activities with IC50 values of 76 and 42 nM, respectively. Both compounds had high selectivity against the other kinases. These D1/CDK4 inhibitors inhibited tumor cell growth, arrested tumor cells at the G1 phase, and inhibited pRb phosphorylation. PMID- 12747777 TI - Novel cationic transport agents for oligonucleotide delivery into primary leukemic cells. AB - Novel cationic compounds forming complexes with oligodeoxyribonucleotides (ODNs) were prepared, and their ability to transport ODNs into cultured primary leukemic cells was tested. Two cationic porphyrin derivatives (2 and 3) were found to be at least 1 order of magnitude more efficient in this respect than commercially available agents. The ODN transporting capacity of novel compounds was dependent on the magnitude and the nature of their positive charges as well as on the porphyrin/ODN molar ratio. Porphyrin-ODN complexes were internalized into cells, and their dissociation was demonstrated by accumulation of fluorescein isothiocyanate-ODN fluorescence in the nucleus. Importantly, porphyrin 3 significantly protected complexed ODN against degradation and efficiently mediated the specific antisense effect on targeted v-Myb expression, resulting in reproducible growth inhibition of treated cells. Low toxicity, serum compatibility, and water solubility of porphyrin 3 make this compound a promising novel tool for modulation of gene expression in primary leukemic cells. PMID- 12747776 TI - 3D QSAR analyses-guided rational design of novel ligands for the (alpha4)2(beta2)3 nicotinic acetylcholine receptor. AB - Three-dimensional quantitative structure-activity relationship methods, the comparative molecular field analysis (CoMFA) and the comparative molecular similarity indices analysis (CoMSIA), were applied using a training set of 45 ligands of the (alpha4)2(beta2)3 nicotinic acetylcholine receptor (nAChR). All compounds are related to (-)-epibatidine, (-)-cytisine, (+)-anatoxin-a, and (-) ferruginine, and additionally, novel diazabicyclo[4.2.1]nonane- and quinuclidin-2 ene-based structures were included. Their biological data have been determined by utilizing the same experimental protocol. Statistically reliable models of good predictive power (CoMFA r2 = 0.928, q2 = 0.692, no. of components = 3; CoMSIA r2 = 0.899, q2 = 0.701, no. of components = 3) were achieved. The results obtained were graphically interpreted in terms of field contribution maps. Hence, physicochemical determinants of binding, such as steric and electrostatic and, for the first time, hydrophobic, hydrogen bond donor, and hydrogen bond acceptor properties, were mapped back onto the molecular structures of a set of nAChR modulators. In particular, changes in the binding affinity of the modulators as a result of modifications in the aromatic ring systems could be rationalized by the steric, electrostatic, hydrophobic, and hydrogen bond acceptor properties. These results were used to guide the rational design of new nAChR ligands such as 48-52 and 54, which were subsequently synthesized for the first time and tested. Key steps of our synthetic approaches were successfully applied Stille and Suzuki cross-coupling reactions. Predictive r2 values of 0.614 and 0.660 for CoMFA and CoMSIA, respectively, obtained for 22 in part previously unknown ligands for the (alpha4)2(beta2)3 subtype, demonstrate the high quality of the 3D QSAR models. PMID- 12747778 TI - Restricted conformation analogues of an anthelmintic cyclodepsipeptide. AB - Six analogues of the anthelmintic cyclodepsipeptide PF1022A were prepared, each containing a small ring fused to the macrocycle to restrict the number of conformations the larger ring can adopt. It was anticipated that such conformational changes could lead to enhanced biological activity and selectivity. The analogues form two series of three members each. In one series, a carbon-based molecular bridge joins the methyl of a leucine residue with the methyl of its closest lactic acid residue to form five-, six-, and seven-membered lactam rings. In the second series, a leucine residue is replaced with five-, six , and seven-membered nitrogen heterocycles. Decreasing the size of the small ring in the lactam series increasingly distorts the macrocycle and consistently decreases activity relative to PF1022A. In the leucine series, a similar trend is observed. Molecular modeling of PF1022A along with the analogues described herein suggests that the ability to exist in a highly symmetrical conformational state is a necessary condition for biological activity. PMID- 12747779 TI - Calculation of the binding affinity of beta-secretase inhibitors using the linear interaction energy method. AB - It has been shown that the rate-limiting step in the production of beta-amyloid peptide (Abeta) is the proteolytric cleavage of the membrane-bound beta-amyloid precursor protein (APP) by beta-secretase (BACE). Since the accumulation of Abeta has been implicated as one of the key events in the progression of Alzheimer's disease, BACE has become an important therapeutic target. Recently, two crystal structures of BACE cocrystallized with the inhibitors OM99-2 and OM00-3 were published by Tang and co-workers. In addition, the Ghosh group has published binding data on a series of inhibitors based on their initial lead, OM99-2. Using this set as a basis, we have developed a model for the binding affinity of these ligands to BACE using the linear interaction energy method. The best binding affinity model for the full set of ligands had a RMSD of 1.10 kcal/mol. The best model excluding the two charged ligands had a RMSD of 0.87 kcal/mol. PMID- 12747780 TI - Binding structures and potencies of oxidosqualene cyclase inhibitors with the homologous squalene-hopene cyclase. AB - The binding structures of 11 human oxidosqualene cyclase inhibitors designed as cholesterol-lowering agents were determined for the squalene-hopene cyclase from Alicyclobacillus acidocaldarius, which is the only structurally known homologue of the human enzyme. The complexes were produced by cocrystallization, and the structures were elucidated by X-ray diffraction analyses. All inhibitors were bound in the large active center cavity. The detailed binding structures are presented and discussed in the light of the IC50 values of these 11 as well as 17 other inhibitors. They provide a consistent picture for the inhibition of the bacterial enzyme and can be used to adjust and improve homology models of the human enzyme. The detailed active center structures of the two enzymes are too different to show an IC50 correlation. PMID- 12747781 TI - Discovery and structure-activity relationship of oxalylarylaminobenzoic acids as inhibitors of protein tyrosine phosphatase 1B. AB - Protein Tyrosine phosphatase 1B (PTP1B) has been implicated as a key negative regulator of both insulin and leptin signaling pathways. Using an NMR-based screening approach with 15N- and 13C-labeled PTP1B, we have identified 2,3 dimethylphenyloxalylaminobenzoic acid (1) as a general, reversible, and competitive PTPase inhibitor. Structure-based approach guided by X-ray crystallography facilitated the development of 1 into a novel series of potent and selective PTP1B inhibitors occupying both the catalytic site and a portion of the noncatalytic, second phosphotyrosine binding site. Interestingly, oral biovailability has been observed in rats for some compounds. Furthermore, we demonstrated in vivo plasma glucose lowering effects with compound 12d in ob/ob mice. PMID- 12747782 TI - Structure-activity relationships of dynorphin a analogues modified in the address sequence. AB - The peptide [Pro3]Dyn A(1-11)-NH2 2 exhibits high affinity (K(i) = 2.4 nM) and over 2000-fold selectivity for the opioid receptor. Stepwise removal of the C terminal residues from this ligand demonstrated that its positively charged Arg residues, particularly Arg6 and Arg7, were crucial for binding to the kappa receptor. Analogues shorter than seven amino acids lacked significant affinity for opioid receptors. Comparison with a series of truncated analogues of Dyn A showed that the relative losses in binding potency differed only slightly between the two series. The neutral residues Ile8 and Pro10 could be removed without significant loss in affinity for the kappa receptor. Their replacement, in the Pro3 analogue, with additional Arg residues led to analogues with improved kappa affinity (e.g., [Pro3,Arg8]Dyn A(1-11)-NH2 20: K(i)(kappa) = 0.44 nM). This type of modification did not compromise the high kappa selectivity of the Pro3 analogues. These findings support the view that a negatively charged domain in the putative second extracellular loop of the kappa receptor selectively recognizes residues 6-11 of dynorphin through electrostatic interactions. As with parent compound 2, analogue 20 and related compounds displayed kappa antagonist properties. PMID- 12747783 TI - Rational design and synthesis of an orally active indolopyridone as a novel conformationally constrained cannabinoid ligand possessing antiinflammatory properties. AB - A series of unique indazoles and pyridoindolones have been rationally designed and synthesized as novel classes of cannabinoid ligands based on a proposed bioactive amide conformation. This has led to the discovery of the novel indolopyridone 3a as a conformationally constrained cannabinoid ligand that displays high affinity for the CB2 receptor (K(i)(CB2) = 1.0 nM) and possesses antiinflammatory properties when administered orally in an in vivo murine inflammation model. PMID- 12747784 TI - Synthesis of chiral 1-[omega-(4-chlorophenoxy)alkyl]-4-methylpiperidines and their biological evaluation at sigma1, sigma2, and sterol delta8-delta7 isomerase sites. AB - Sumitomo's patented sigma ligand 1-[3-(4-chlorophenoxy)propyl]-4-methylpiperidine (15), which has been claimed as agent for CNS disorders and neuropathies, and its lower homologue 12 were prepared along with related chiral (4 chlorophenoxy)alkylpiperidines. They were tested at sigma1, sigma2, and sterol Delta8-Delta7 isomerase (SI) sites by in vitro radioligand binding assays, to evaluate the influence of a chiral center in the alkyl chain on the selective sigma(1) binding relative to other sigma family sites. Generally high sigma1-site affinities were found, so that the chirality introduced by a methyl substitution resulted in slight differences. Nevertheless, the shorter oxyethylenic chain was beneficial to increase sigma1 selectivity. However, the (-)-(S)-4-methyl-1-[2-(4 chlorophenoxy)-1-methylethyl]piperidine ((-)-(S)-17) reached the highest sigma1 affinity (K(i) = 0.34 nM) and the best selectivity relative to the sigma2 site (547-fold). Compound (-)-(S)-17 displayed also a moderate selectivity (11-fold) relative to the SI site. PMID- 12747785 TI - Modulation of P-glycoprotein-mediated multidrug resistance by flavonoid derivatives and analogues. AB - Flavonoid derivatives were synthesized and tested for their ability to modulate P glycoprotein (Pgp)-mediated multidrug resistance (MDR) in vitro. These compounds belong to various flavonoid subclasses, namely: chromones, azaisoflavones, and aurones. Among the investigated compounds, three showed potent reversing activity. 2-(4-methylpiperazin-1-ylcarbonyl)-5-hydroxychromone (4a), 5,7 dimethoxy-3-phenyl-4-quinolone (5), and 4,6-dimethoxyaurone (6) potentiated daunorubicin cytotoxicity on resistant K562 cells. They were also able to increase the intracellular accumulation of rhodamine-123, a fluorescent molecule which acts as a probe of P-glycoprotein-mediated MDR. This suggests that these compounds act, at least in part, by inhibiting P-glycoprotein activity. The most active compound, 5-hydroxy-2-(4-methylpiperazin-1-ylcarbonyl)chromone (4a) was found to be a powerful reversal agent, more potent than cyclosporin A, used as the reference molecule. No effect was observed on MRP transport nor on cell proliferation. Little apoptosis was induced on K562S cells with 4a compared to K562R, probably due to the extrusion of the compound by Pgp. PMID- 12747786 TI - Unsymmetrical DNA cross-linking agents: combination of the CBI and PBD pharmacophores. AB - A set of 10 compounds, each combining the seco-1,2,9,9a tetrahydrocyclopropa[c]benz[e]indol-4-one (seco-CBI) and pyrrolo[2,1 c][1,4]benzodiazepine (PBD) pharmacophores, was designed and prepared. These compounds were anticipated to cross-link between N3 of adenine and N2 of guanine in the minor groove of DNA. The compounds, which differ in the chain length separating the two alkylation subunits, and the configuration of the CBI portion, showed great variation in cellular toxicity (over 4 orders of magnitude in a cell line panel) with the most potent example exhibiting IC50s in the pM range. Cytotoxicity correlated with the ability of the compounds to cross-link naked DNA. Cross-linking was also observed in living cells, at much lower concentrations than for a related symmetrical PBD dimer. A thermal cleavage assay was used to assess sequence selectivity, demonstrating that the CBI portion controlled the alkylation sites, while the PBD substituent increased the overall efficiency of alkylation. Several compounds were tested for in vivo activity using a tumor growth delay assay against WiDr human colon carcinoma xenografts, with one compound (the most cytotoxic and most efficient cross-linker) showing a statistically significant increase in survival time following a single iv dose. PMID- 12747787 TI - Discovery of a simple picomolar inhibitor of cholesteryl ester transfer protein. AB - A novel series of substituted N-[3-(1,1,2,2-tetrafluoroethoxy)benzyl]-N-(3 phenoxyphenyl)-trifluoro-3-amino-2-propanols is described which potently and reversibly inhibit cholesteryl ester transfer protein (CETP). Starting from the initial lead 1, various substituents were introduced into the 3-phenoxyaniline group to optimize the relative activity for inhibition of the CETP-mediated transfer of [3H]-cholesteryl ester from HDL donor particles to LDL acceptor particles either in buffer or in human serum. The better inhibitors in the buffer assay clustered among compounds in which the phenoxy group was substituted at the 3, 4, or 5 positions. In general, small lipophilic alkyl, haloalkyl, haloalkoxy, and halogen moieties increased potency relative to 1, while analogues containing electron-donating or hydrogen bond accepting groups exhibited lower potency. Compounds with polar or strong electron-withdrawing groups also displayed lower potency. Replacement of the phenoxy ring in 1 with either simple aliphatic or cycloalkyl ethers as well as basic heteroaryloxy groups led to reduced potency. From the better compounds, a representative series 4a-i was prepared as the chirally pure R(+) enantiomers, and from these, the 4-chloro-3-ethylphenoxy analogue was identified as a potent inhibitor of CETP activity in buffer (4a, IC50 0.77 nM, 59 nM in human serum). The simple R(+) enantiomer 4a represents the most potent acyclic CETP inhibitor reported. The chiral synthesis and biochemical characterization of 4a are reported along with its preliminary pharmacological assessment in animals. PMID- 12747789 TI - Metal coordination-based inhibitors of adenylyl cyclase: novel potent P-site antagonists. AB - The adenylyl cyclases (ACs) are a family of intracellular enzymes associated with signal transduction by virtue of their ability to convert ATP to cAMP. The catalytic mechanism of this transformation proceeds through initial binding of ATP to the so-called purine binding site (P-site) of the enzyme followed by metal mediated cyclization with loss of pyrophosphate. Crystallographic analysis of ACs with known inhibitors reveals the presence of two metals in the active site. Presently, nine isoforms of adenylyl cyclase are known, and unique isoform combinations are expressed in a tissue-specific manner. The development of isoform-specific inhibitors of adenylyl cyclase may prove to be a useful strategy toward the design of unique signal transduction inhibitors. To develop novel AC inhibitors, we have chosen an approach to inhibitor design utilizing an adenine ring system joined to a metal-coordinating hydroxamic acid via various linkers. Previous work in our group has validated this approach and identified novel inhibitors that possess an adenine ring joined to a metal-coordinating hydroxamic acid through flexible acyclic linkers (Levy, D. E., et al. Bioorg. Med. Chem. Lett. 2002, 12, 3085-3088). Subsequent studies have focused on the introduction of conformational restrictions into the tether of the inhibitors with the goal of increasing potency (Levy, D. E., et al. Bioorg. Med. Chem. Lett. 2002, 12, 3089 3092). Building upon the favorable spatial positioning of the adenine and hydroxamate groups coupled with potentially favorable entropic factors, the unit joining the carbocycle to the hydroxamate was explored further and a stereochemical-based SAR was elucidated, leading to a new series of highly potent AC inhibitors. PMID- 12747788 TI - Imidazoline binding sites (IBS) profile modulation: key role of the bridge in determining I1-IBS or I2-IBS selectivity within a series of 2 phenoxymethylimidazoline analogues. AB - The alpha- and beta-methyl derivatives of 2-phenylethylimidazoline (compounds 7 and 8) and the corresponding enantiomers were prepared and tested with the purpose of studying the role played by the ethylene bridge in modulating I(1)- and I(2)-IBS selectivity. The alpha-methylation appeared to be extremely critical regarding the affinity and selectivity for the I1-IBS subtypes (I1/I2 = 186 for imidazoline 7) and the stereospecificity of interaction (eudismic ratio (S)-(-) 7/(R)-(+)-7 = 5888). Instead, even if in a more limited fashion, the -methylation tended toward I2-IBS selectivity (I2/I1 = 50 for imidazoline 8). The unsubstituted compound 4 (I2/I1 = 1479) proved to be considerably more potent and selective with respect to I2-IBS subtypes. PMID- 12747790 TI - Carbonic anhydrase inhibitors. Inhibition of tumor-associated isozyme IX by halogenosulfanilamide and halogenophenylaminobenzolamide derivatives. AB - Two series of halogenated sulfonamides have been prepared. The first consists of mono/dihalogenated sulfanilamides, whereas the second one consists of the mono/dihalogenated aminobenzolamides, incorporating equal or different halogens (F, Cl, Br, and I). These sulfonamides have been synthesized from the corresponding anilines by acetylation (protection of the amino group), chlorosulfonylation, followed either by amidation, or reaction with 5-amino-1,3,4 thiadiazole-2-sulfonamide (and eventually deacetylation). All these compounds, together with the six clinically used sulfonamide inhibitors (acetazolamide, methazolamide, ethoxzolamide, dichlorophenamide, dorzolamide, and brinzolamide) were investigated as inhibitors of the transmembrane, tumor-associated isozyme carbonic anhydrase (CA) IX. Inhibition data against the classical, physiologically relevant isozymes I, II, and IV were also obtained. CA IX shows an inhibition profile which is generally completely different from those of isozymes I, II, and IV, with potent inhibitors (inhibition constants in the range of 12-40 nM) among both simple aromatic (such as 3-fluoro-5-chloro-4 aminobenzenesulfonamide) as well as heterocyclic compounds (such as acetazolamide, methazolamide, 5-amino-1,3,4-thiadiazole-2-sulfonamide, aminobenzolamide, and dihalogenated aminobenzolamides). This first detailed CA IX inhibition study revealed many interesting leads, suggesting the possibility to design even more potent and eventually CA IX-selective inhibitors, with putative applications as antitumor agents. PMID- 12747791 TI - Carbonic anhydrase inhibitors. Inhibition of cytosolic isozymes I and II and transmembrane, tumor-associated isozyme IX with sulfamates including EMATE also acting as steroid sulfatase inhibitors. AB - A series of sulfamates or bis-sulfamates incorporating aliphatic, aromatic, polycyclic (steroidal), and sugar moieties in their molecules has been synthesized and assayed as inhibitors of the zinc enzyme carbonic anhydrase (CA), and more precisely of the cytosolic isozymes CA I andII, and the transmembrane, tumor-associated isozymes CA IX. Some of these compounds were previously reported to act as inhibitors of steroid sulfatases, among which estrone sulfatase (ES) and dehydroepiandrosterone sulfatase (DHEAS) are the key therapeutic targets for estrogen-dependent tumors. Very potent (nanomolar) inhibitors were detected against the three investigated CA isozymes. Best CA I inhibitors were phenylsulfamate and some of its 4-halogeno derivatives, as well as the aliphatic compound n-octyl sulfamate. Against CA II, low nanomolar inhibitors (1.1-5 nM) were phenylsulfamate and some of its 4-halogeno/nitro derivatives, n-octyl sulfamate, and estradiol 3,17beta-disulfamate among others. All the investigated sulfamates showed efficient CA IX inhibitory properties, with inhibition constants in the range of 18-63 nM. The best CA IX inhibitor detected so far was 4-chlorophenylsulfamate. These data are critical for the design of novel antitumor properties, mainly for hypoxic tumors that overexpress CA IX, which are nonresponsive to radiation or chemotherapy. The antitumor properties of the ES/DHEAS inhibitors in clinical trials may on the other hand also be due to their potent inhibitory properties of CA isozymes involved in tumorigenicity, such as CA II and CA IX. PMID- 12747793 TI - C(8) substituted 1-azabicyclo[3.3.1]non-3-enes and C(8) substituted 1 azabicyclo[3.3.1]nonan-4-ones: novel muscarinic receptor antagonists. AB - Expedient syntheses of C(8) substituted 1-azabicyclo[3.3.1]non-3-enes and C(8) substituted 1-azabicyclo[3.3.1]nonan-4-ones are reported to begin with 2,5 disubstituted pyridines. Catalytic reduction of the pyridine to the piperidine followed by treatment with ethyl acrylate and Dieckmann cyclization gave diastereomeric mixtures of C(8) substituted 3-ethoxycarbonyl-4-hydroxy-1 azabicyclo[3.3.1]non-3-enes, which were separable by chromatography. We found that the catalytic reduction (PtO2, H2) procedure provided the cis-substituted piperidine but that pyridine reduction was accompanied by competitive cleavage of the C(2) pyridyl substituent. Accordingly, an alternative route was devised that afforded a diastereomeric mixture of the cis- and trans-2,5-disubstituted piperidine. Treatment of the substituted pyridine with m-CPBA gave the pyridine N oxide, which was reduced to the piperidine by sequential reduction with ammonium formate in the presence of Pd-C followed by NaBH3CN. Addition of ethyl acrylate completed the synthesis of the substituted piperidine. The overall four-step reaction gave higher yields (57%) than the two-step procedure (13%) with little cleavage of the C(2) pyridyl substituent. Acid decarboxylation of the bicyclo[3.3.1]non-3-enes provided the C(8) substituted 1-azabicyclo[3.3.1]nonan-4 ones. Structural studies revealed diagnostic 13C NMR signals that permit assignment of the orientation of the C(8) substituent. Pharmacological investigations documented that 3-ethoxycarbonyl-4-hydroxy-1-azabicyclo[3.3.1]non 3-enes efficiently bind to the human M1-M5 muscarinic receptors and function as antagonists. We observed that exo-8-benzyloxymethyl-3-ethoxycarbonyl-4-hydroxy-1 azabicyclo[3.3.1]non-3-ene (3) displayed the highest affinity, exhibiting Ki values at all five muscarinic receptors that were approximately 10-50 times lower than carbachol and approximately 30-230 times lower than arecoline. Receptor selectivity was observed for 3. Compound 3 contained two different pharmacophores found in many muscarinic receptor ligands, and preliminary findings indicated the importance of both structural elements for maximal activity. Compound 3 serves as a novel lead compound for further drug development. PMID- 12747792 TI - Interaction of cis-(6-benzhydrylpiperidin-3-yl)benzylamine analogues with monoamine transporters: structure-activity relationship study of structurally constrained 3,6-disubstituted piperidine analogues of (2,2-diphenylethyl)-[1-(4 fluorobenzyl)piperidin-4-ylmethyl]amine. AB - To explore structure-activity relationships (SAR) of a novel conformationally constrained lead cis-3,6-disubstituted piperidine derivative derived from (2,2 diphenylethyl)-[1-(4-fluorobenzyl)piperidine-4-ylmethyl]amine (I), a series of compounds was synthesized by derivatizing the exocyclic N-atom at the 3-position of the lead. This study led to the formation of substituted phenyl and heterocyclic derivatives. All novel compounds were tested for their affinity at the dopamine transporter (DAT), serotonin transporter (SERT), and norepinephrine transporter (NET) in the brain by measuring their potency in competing for the binding of [3H]WIN 35 428, [3H]citalopram, and [3H]nisoxetine, respectively. Selected compounds were also evaluated for their activity in inhibiting the uptake of [3H]DA. The SAR results demonstrated that the nature of substitutions on the phenyl ring is important in activity at the DAT with the presence of an electron-withdrawing group having the maximum effect on potency. Replacement of the phenyl ring in the benzyl group by heterocyclic moieties resulted in the development of compounds with moderate activity for the DAT. Two most potent racemic compounds were separated by a diastereoisomeric separation procedure, and differential affinities were observed for the enantiomers. Absolute configuration of the enantiomers was obtained unambiguously by X-ray crystal structural study. One of the enantiomers, compound S,S-(-)-19a, exhibited the highest potency for the DAT (IC50 = 11.3 nM) among all the compounds tested and was as potent as GBR 12909 (1-[2-[bis(4-fluorophenyl)methoxy]ethyl]-4-(3-phenylpropyl)piperazine). However, the compound (-)-19a was more selective than GBR 12909 in binding to the DAT compared with binding to the SERT and NET. The present results establish the newly developed 3,6-disubstituted piperidine derivatives as a novel template for high-affinity inhibitors of DAT. Structurally these molecules are more constrained compared to our earlier flexible piperidine molecules and, thus, should provide more insights about their bioactive conformations. PMID- 12747795 TI - Determination of conformer-specific partition coefficients in octanol/water systems. AB - The first conformer-specific experimental partition coefficients are presented for octanol/water, the most widespread solvent system to predict lipophilicity of drugs. Rotamer populations in octanol and water were elucidated from 1H NMR vicinal coupling constants and were combined with classical partition coefficients to obtain the conformer-specific ones. Feasibility of the determination of conformer-specific partition coefficients is exemplified on amphetamine and clenbuterol, two flexible drug molecules. Partition capacities of the amphetamine rotamers have been proven to be essentially equal. The conformers of clenbuterol, however, have been found to be greatly different in partition properties, which could be interpreted in terms of intramolecular interactions between the vicinal polar sites and the solvent-accessibility of the groups. The conformers could be put into order of their membrane-influx and -outflow propensities. Deviations between experimental and calculated log P values could also be interpreted in view of the species-specific partition coefficients. PMID- 12747794 TI - Identification of a novel, selective GABA(A) alpha5 receptor inverse agonist which enhances cognition. AB - In pursuit of a GABA(A) alpha5-subtype-selective inverse agonist to enhance cognition, a series of 6,7-dihydro-2-benzothiophen-4(5H)-ones has been identified as a novel class of GABA(A) receptor ligands. These thiophenes have higher binding affinity for the GABA(A) alpha5 receptor subtype compared to the GABA(A) alpha1, alpha2, and alpha3 subtypes, and several analogues exhibit high GABA(A) alpha5 receptor inverse agonism. 6,6-Dimethyl-3-(2-hydroxyethyl)thio-1-(thiazol-2 yl)-6,7-dihydro-2-benzothiophen-4(5H)-one (43) has been identified as a full inverse agonist at the GABA(A) alpha5 receptor and is functionally selective over the other major GABA(A) receptor subtypes. 43 readily penetrates into the CNS to give selective occupancy of GABA(A) alpha5 receptors. In addition, 43 enhances cognitive performance in rats in the delayed 'matching-to-place' Morris water maze test-a hippocampal-dependent memory task-without the convulsant or proconvulsant activity associated with nonselective, GABA(A) receptor inverse agonists. PMID- 12747796 TI - Design, synthesis, and pharmacology of a highly subtype-selective GluR1/2 agonist, (RS)-2-amino-3-(4-chloro-3-hydroxy-5-isoxazolyl)propionic acid (Cl HIBO). AB - On the basis of structural studies, chloro-homoibotenic acid (Cl-HIBO) was designed and synthesized. Cl-HIBO was characterized in binding and electrophysiology experiments on native and cloned subtypes of GluRs. Electrophysiological selectivities ranged from 275 to 1600 for GluR1/2 over GluR3/4. The potent AMPA receptor activity was strongly desensitizing and the neurotoxicity similar to AMPA. Thus, Cl-HIBO is the most subtype selective agonist reported to date on GluR1/2, and offers a new standard for selectively studying subtypes of AMPA receptors. PMID- 12747797 TI - Distorting malaria peptide backbone structure to enable fitting into MHC class II molecules renders modified peptides immunogenic and protective. AB - The conserved, nonantigenic, nonimmunogenic malaria Merozoite Surface Protein-2 peptide 1, having high affinity for red blood cells, was rendered immunogenic and protective in Aotus monkeys by specifically changing some critical residues. The NMR structure revealed a switch from classical type III' into distorted III' and III beta turns in the protective peptides. These changes may lead to a better fit into the Aotus MHC class II human HLA-DRbeta1 12 molecule equivalent, thus activating the immune system. PMID- 12747798 TI - Nitro and amino substitution in the D-ring of 5-(2-dimethylaminoethyl)- 2,3 methylenedioxy-5H-dibenzo[c,h][1,6]naphthyridin-6-ones: effect on topoisomerase-I targeting activity and cytotoxicity. AB - 5H-8,9-dimethoxy-5-(2-N,N-dimethylaminoethyl)-2,3 methylenedioxydibenzo[c,h][1,6]naphthyridin-6-one exhibits potent TOP1-targeting activity and pronounced antitumor activity. It was hypothesized that replacement of the two methoxyl groups with a nitro substituent would allow for retention of similar activity. In this study 8-, 9-, and 10-nitro-5H-2,3-methylenedioxy-5-(2 N,N-dimethylaminoethyl)dibenzo[c,h][1,6]naphthyridin-6-one and their amino derivatives were synthesized and assessed for their relative TOP1-targeting activity and cytotoxicity. In the case of both the 8- and 9-nitro analogues, their TOP1-targeting activity and cytotoxicity are greater than that of camptothecin and comparable to that of 5H-8,9-dimethoxy-5-(2-N,N dimethylaminoethyl)-2,3-methylenedioxydibenzo[c,h][1,6]naphthyridin-6-one. PMID- 12747799 TI - Higher prevalence of Chlamydia pneumoniae seropositivity in Finnish twins compared with co-twins living in Sweden: relationships with markers of subclinical atherosclerosis. AB - Mortality rates from coronary heart disease are lower in Swedish men and among Finnish migrants who have lived in Sweden for over 20 years than in men living in Finland. Sero-epidemiological studies, investigations of atheromatous plaque specimens, in vitro animal models and anti-chlamydial antibiotic trials have given support to the hypothesis that Chlamydia pneumoniae (Cpn) has a role in atherosclerosis. We investigated whether men with a similar genetic background but living permanently in Finland or Sweden have differences in the prevalence of Cpn seropositivity, and whether chronic Cpn infection is associated with markers of subclinical atherosclerosis. We measured anti-Cpn antibodies and ultrasonographic markers of subclinical atherosclerosis, including carotid intima media thickness, carotid artery compliance and brachial artery flow-mediated dilatation, in a population of 76 migrant-discordant male twin pairs (152 men). The number of men with seropositivity to Cpn infection (defined as IgA>/=1:64 and IgG>/=1:128) was greater in Finland than in Sweden (21.5% compared with 10.5%; P =0.046). Cpn seropositivity accompanied by elevated C-reactive protein (CRP) levels (>1 mg/l) was associated with attenuated brachial artery flow-mediated dilatation (3.3+/-0.3%, compared with 5.5+/-0.4% in men with no signs of Cpn infection; P <0.001).Thus, among Finnish twin brothers discordant for migration to Sweden, the prevalence of Cpn seropositivity is higher for those living in Finland, and men with Cpn seropositivity combined with elevated CRP levels had attenuated endothelial function. These findings offer insight into the mechanism whereby chronic Cpn infection may increase the risk of coronary heart disease. PMID- 12747800 TI - P2X7 receptor activates extracellular signal-regulated kinases ERK1 and ERK2 independently of Ca2+ influx. AB - P2X7 nucleotide receptors modulate a spectrum of cellular events in various cells including epithelia, such as exocrine pancreas. Although the pharmacology and channel properties of the P2X7 receptors have been studied intensively, signal transduction pathways are relatively unknown. In this study we applied a heterologous expression system of rat P2X7 receptors in HEK-293 cells. We followed the receptor expression and function using the enhanced green fluorescent protein (EGFP) tag, activation of intracellular proteins and increases in cellular Ca2+. EGFP-P2X7 receptors localized to the plasma membrane, clusters within the membrane and intracellularly. Stimulation of P2X7 receptors in HEK-293 cells led to an activation of extracellular signal-regulated kinases ERK1 and ERK2 and this activation was seen after just 1 min of stimulation with ATP. Using C- and N-terminal P2X7-receptor mutants we show that the N-terminus is important in activation of ERKs, whereas deletion of the last 230 amino acids in the C-terminus did not effect ERK activation. On the other hand, Ca2+ entry was impaired in C-terminal but not in N-terminal mutants. In cell suspensions prepared from rat pancreas we show that P2X7 receptors also activate ERK1 and ERK2, indicating that these signalling pathways are also turned on in native epithelium. PMID- 12747802 TI - ATP-binding sites in brain p97/VCP (valosin-containing protein), a multifunctional AAA ATPase. AB - VCP (valosin-containing protein) or p97 is a member of the AAA family (ATPases associated with a variety of cellular activities family), a diverse group of proteins sharing a key conserved AAA module containing duplicate putative ATP binding sites. Although the functions of the AAA family are related to their putative ATP-binding sites, the binding of ATP to these sites has not yet been demonstrated. In the present study, the ATP-binding site(s) of brain VCP was characterized using the photoreactive ATP analogue, BzATP [3'- O -(4 benzoylbenzoyl)ATP]. Photo-activation of Bz-[alpha-(32)P]ATP resulted in its covalent binding to a 97-kDa purified soluble or membrane-associated protein, identified by amino acid sequencing as VCP. Bz-[alpha-(32)P]ATP covalently bound to the purified homo-hexameric VCP with an apparent high affinity (74-111 nM). A molar stoichiometry of 2.23+/-0.14 BzATP bound per homo-hexameric VCP (n =6) was determined using different methods for analysis of radiolabelling and protein determination. Nucleotides inhibited the binding of Bz-[alpha-(32)P]ATP to VCP with the following efficiency: BzATP>ATP>ADP>>adenosine 5'-[beta,gamma imido]triphosphate>or=adenosine 5'-[beta,gamma-methylene]triphosphate, whereas AMP, GTP and CTP were ineffective. VCP was observed to possess very low ATPase activity, with nucleotide specificity similar to that for BzATP binding. Conformational changes induced by an alternating site mechanism for ATP binding are suggested as a molecular mechanism for coupling ATP binding to the diverse activities of the AAA family. PMID- 12747801 TI - Identification of ubiquitination sites on the X-linked inhibitor of apoptosis protein. AB - The execution phase of apoptosis is under the control of members of the inhibitor of apoptosis (IAP) family of zinc finger proteins. Several of these proteins contain a C-terminal RING (really interesting new gene) domain that has been postulated to regulate ubiquitination of themselves or their target proteins, thereby modulating thresholds for apoptosis. We demonstrate that the auto ubiquitination sites of the X-linked IAP (XIAP) are Lys(322) and Lys(328), located in the third baculovirus IAP repeat domain of the protein. Modification of these sites to arginine dramatically reduces ubiquitination of XIAP, but has no measurable effect on the ability of ectopically expressed IAP to rescue cells from two independent apoptotic inducers. Our data firmly locate the auto ubiquitination sites, and raise doubts regarding the importance of this event as a mechanism for regulating the levels of XIAP. PMID- 12747803 TI - The three isoenzymes of human inositol-1,4,5-trisphosphate 3-kinase show specific intracellular localization but comparable Ca2+ responses on transfection in COS-7 cells. AB - Inositol 1,4,5-trisphosphate [Ins(1,4,5) P3] 3-kinase catalyses the phosphorylation of InsP3 to inositol 1,3,4,5-tetrakisphosphate. cDNAs encoding three human isoenzymes of InsP3 3-kinase (A, B and C) have been reported previously [Choi, Kim, Lee, Moon, Sim, Kim, Chung and Rhee (1990) Science 248, 64 66; Dewaste, Pouillon, Moreau, Shears, Takazawa and Erneux (2000) Biochem. J. 352, 343-351; Dewaste, Roymans, Moreau and Erneux (2002) Biochem. Biophys. Res. Commun. 291, 400-405; Takazawa, Perret, Dumont and Erneux (1991) Biochem. Biophys. Res. Commun. 174, 529-535]. The localization of InsP3 3-kinase isoenzymes fused at their N-terminus to the green fluorescent protein has been studied by confocal microscopy. The A isoform appeared to associate with the cytoskeleton, whereas the C isoform was totally cytoplasmic. The B isoform had a more complex localization: it appeared in the plasma membrane, cytoskeleton and in the endoplasmic reticulum. The three human isoenzymes of InsP3 3-kinase can thus be distinguished by their N-terminal sequence, sensitivity to Ca2+/calmodulin and localization on transfection in COS-7 cells. We have compared the cytosolic Ca2+ responses induced by ATP in COS-7 cells transfected with the three isoenzymes. Cells expressing high levels of any of the three isoforms no longer respond to ATP, whereas cells expressing low levels of each enzyme showed a reduced response consisting of one to three Ca2+ spikes in response to 100 microM ATP. These effects were seen only in wild-type InsP3 3-kinase-transfected cells. 3-Kinase-dead mutant cells behaved as vector-transfected cells. The results highlight the potential role of the three isoforms of InsP3 3-kinase as direct InsP3 metabolizing enzymes and direct regulators of Ca2+ responses to extracellular signals. PMID- 12747804 TI - Muscarinic receptor-mediated activation of p70 S6 kinase 1 (S6K1) in 1321N1 astrocytoma cells: permissive role of phosphoinositide 3-kinase. AB - In 1321N1 astrocytoma cells, carbachol stimulation of M3 muscarinic cholinergic receptors, coupled to phospholipase C, evoked a persistent 10-20-fold activation of p70 S6 kinase (S6K1). This response was abolished by chelation of cytosolic Ca2+ and reproduced by the Ca2+ ionophore ionomycin, but was not prevented by down-regulation or inhibition of protein kinase C. Carbachol-stimulated activation and phosphorylation of S6K1 at Thr389 were prevented by rapamycin, an inhibitor of mTOR (mammalian target of rapamycin), or by wortmannin, a phosphoinositide 3-kinase (PI3K) inhibitor. Carbachol also stimulated the phosphorylation of eukaryotic initiation factor 4E-binding protein-1 (4E-BP1), a second mTOR-dependent event, with similar potency to its effect on S6K1. This response was blocked by rapamycin, but was not markedly affected by 100 nM wortmannin, implying separate roles for mTOR and PI3K in S6K1 activation. Wortmannin abolished the carbachol-stimulated rise in PtdIns(3,4,5)P3 and greatly reduced unstimulated levels of this lipid. By contrast, an inhibitor of epidermal growth factor receptor kinase, AG1478, which prevents carbachol-stimulated ErbB3 transactivation, PI3K recruitment and protein kinase B activation in 1321N1 cells, reduced activation of S6K1 by no more than 30%. This effect was overcome by 10 nM insulin, which on its own did not stimulate S6K1, but increased cellular PtdIns(3,4,5)P3 concentrations comparably with carbachol alone. These observations distinguish obligatory roles for mTOR and PI3K in regulating S6K1, but imply that minimal PI3K activity is sufficient to permit stimulation of S6K1 by other activating factors such as increased cytosolic Ca2+ concentrations, which are essential to the muscarinic receptor-mediated response. Moreover, 4E BP1 and hence, presumably, mTOR can be regulated independently of PI3K activation through these mechanisms. PMID- 12747805 TI - Enhanced expression of the gene for beta-glycosidase of Thermus caldophilus GK24 and synthesis of galacto-oligosaccharides by the enzyme. AB - The gene (bglT) encoding Tca beta-glycosidase (Thermus caldophilus GK24 beta glycosidase) was overexpressed under the control of the trp promoter on a high copy-number plasmid, pTRPES, in Escherichia coli W3110. The purified Tca beta glycosidase enzyme was used in a galactosyl-transfer reaction to synthesize galacto-oligosaccharides from lactose. The optimum temperature and pH for the enzyme to synthesize galacto-oligosaccharides from 30% (w/v) lactose were 80 degrees C and 6.0, respectively. The major product of the reaction was a trisaccharide. The thermostable Tca beta-glycosidase produced galacto oligosaccharides efficiently during the hydrolysis of lactose. PMID- 12747806 TI - Aromatase expression and role of estrogens in male gonad : a review. AB - The ability of the testis to convert irreversibly androgens into estrogens is related to the presence of a microsomal enzymatic complex named aromatase, which is composed of a specific glycoprotein, the cytochrome P450 aromatase (P450arom) and an ubiquitous reductase. The aromatase gene is unique in humans and contained 18 exons, 9 of them being translated. In the rat testis we have immunolocalized the P450arom not only in Leydig cells but also in germ cells and especially in elongated spermatids. Related to the stage of germ cell maturation, we have shown that the level of P450arom mRNA transcripts decreases, it is much more abundant in pachytene spermatocytes and round spermatids than in mature germ cells whereas the aromatase activity is 2-4 fold greater in spermatozoa when compared to the younger germ cells. Using a highly specific quantitative competitive RT-PCR method we have evidenced that several factors direct the expression of the aromatase gene in Leydig cells, Sertoli cells, pachytene spermatocytes and round spermatids, and it is obvious that promoter PII is the main one but other promoters could be concerned. In the bank-vole testis we have observed a positive correlation between a fully developed spermatogenesis and a strong immunoreactivity for both P450arom and estrogen receptor beta not only in Sertoli cells but also in pachytene spermatocytes and round spermatids. Our recent data obtained from ejaculated human spermatozoa demonstrate the presence of aromatase both in terms of mRNA and protein, and in addition, we suggest that aromatase could be involved in the acquisition of sperm motility. Indeed in men the congenital aromatase deficiency is associated with severe bone maturation problems and sterility. Together with the widespread distribution of estrogen receptors in testicular cells these data clearly show that estrogens play a physiological role in the regulation of spermatogenesis in mammals. PMID- 12747807 TI - The Aging Males' Symptoms (AMS) scale: update and compilation of international versions. AB - BACKGROUND: The interest of clinical research in aging males increased in recent years and thereby the interest to measure health-related quality of life (HRQoL) and symptoms of aging men. The Aging Males' Symptoms scale (AMS) became the most commonly used scale to measure HRQoL and symptoms in aging males in many countries worldwide. The aim of this paper is to review the current state of the instrument particularly concerning versions of the scale in different languages in the light of the quality of the translation process. AMS VERSIONS AVAILABLE: Most of the translations were performed following international methodological recommendations for linguistic & cultural adaptation of HRQoL instruments. Mainly the English version was used as source language for the translation into Dutch, Spanish, Portuguese, Italian, Swedish, and Japanese (attached as additional PDF files). Preliminary versions that were derived only from forward translations are of secondary quality and available in Finnish, Flemish, and Russian. It is recommended to complete the translation process for the latter languages before using them in international studies. TRANSLATIONS IN PROCESS: The AMS scale is in the process of consensus finding of two existing French versions, and the versions in the Korean, Thai, and Indonesian languages have not yet been completed in the translation process. CONCLUSION: The AMS scale is obviously a valuable tool for assessing health related quality of life in aging men, because it is used worldwide. It is a standardized scale according to psychometric norms. Most of the currently available language versions were translated following international standards for linguistic and cultural translation of quality of life scales. Assistance is offered to help interested parties in the translation process. PMID- 12747808 TI - Interfering with TGFbeta-induced Smad3 nuclear accumulation differentially affects TGFbeta-dependent gene expression. AB - BACKGROUND: Transforming growth factor-beta (TGFbeta) plays an important role in late-stage carcinogenesis by stimulating invasive behavior of cancer cells, promoting neo-angiogenesis and by helping cancer cells to escape surveillance by the immune system. It also supports colonization of the bone by metastatic breast cancer cells by increasing expression of osteolytic parathyroid hormone-related protein (PTHrP). Interfering with TGFbeta signalling may thus weaken the malignant properties of cancer cells. We investigated to what extent two inhibitors, SB-202190 and SB-203580, interfere with TGFbeta-signalling in invasive MDA-MB-231 breast cancer cells. These compounds, formerly used as p38 MAPK-specific inhibitors, were recently also demonstrated to inhibit TGFbeta type I receptor kinase. RESULTS: Our results show that these inhibitors delay the onset of TGFbeta-induced nuclear accumulation of Smad3 and reduces its amplitude. This effect was accompanied by a strong reduction in TGFbeta-responsivess of the slow-responder genes pthrp, pai-1 and upa, while the reactivity of the fast responder gene smad7 to TGFbeta remained almost unchanged. Neither was the TGFbeta response of the fast-responder ese-1/esx gene, whose expression we found to be strongly downregulated by TGFbeta, affected by the inhibitors. CONCLUSION: The data show that SB-202190 and SB-203580 suppress TGFbeta-dependent activation of genes that are important for the acquisition of invasive behavior, while having no effect on the expression of the natural TGFbeta inhibitor Smad7. This suggests that these compounds are potent inhibitors of malignant behavior of cancer cells. PMID- 12747809 TI - Calcitonin receptor gene expression in K562 chronic myelogenous leukemic cells. AB - BACKGROUND: The peptide hormone calcitonin (CT) can significantly effect the proliferation rate of CT receptor (CTR) positive human cancer cells. We wish to identify additional human cancers expressing CTRs and assay the effects of CT on their growth rates and signal transduction pathways. RESULTS: The expression of the human calcitonin receptor (hCTR) gene in the chronic myelogenous leukemia cell line K562 was examined. RT-PCR on total RNA extracted from K562 cells detected the presence of hCTR mRNA. Further analysis demonstrated that multiple hCTR isoforms were present. Incubation of K562 cells with salmon calcitonin (sCT), but not amylin, caused an increase in intracellular levels of cAMP similar to that induced by forskolin treatment. We further demonstrated that butyrate induced erythroid differentiation of K562 cells caused a significant decrease in hCTR mRNA levels. However, phorbol myristate acetate (PMA) induced megakaryocytic differentiation of these cells had no significant effect on hCTR mRNA levels. We demonstrated that exposure to various concentrations of sCT had no effect on the cellular proliferation of K562 cells in vitro. CONCLUSION: Chronic myelogenous k562 cells express multiple CTR isoforms. However, CT does not effect K562 proliferation rates. It is likely that the small increase in intracellular levels of cAMP following CT treatment is not sufficient to interfere with cellular growth. PMID- 12747810 TI - Quality of life and metabolic status in mildly depressed women with type 2 diabetes treated with paroxetine: a single-blind randomised placebo controlled trial. AB - BACKGROUND: Depression is prevalent in people with type 2 diabetes and affects both glycemic control and overall quality of life. The aim of this trial was to evaluate the effect of the antidepressant paroxetine on metabolic control, quality of life and mental well-being in mildly depressed women with type 2 diabetes. METHODS: We randomised 15 mildly depressed women with non-optimally controlled type 2 diabetes to a 10-week single-blind treatment with either paroxetine 20 mg per day or placebo. Primary efficacy measurements were glycemic control and quality of life. Glycosylated hemoglobin A1c (GHbA1c) was used as a measure of glycemic control. Quality of life was evaluated using RAND-36. Mental state was assessed using two clinician-rated scoring instruments, Hamilton's Anxiety Scale (HAM-A) and Montgomery-Asberg's Depression Rating Scale (MADRS), and a patient-rated scoring instrument, Beck's Depression Inventory (BDI). RESULTS: At the end of the study no significant difference between groups in improvement of quality of life was found. A trend towards a superior improvement in glycemic control was found in the paroxetine group (p = 0.08). A superior increase in sex-hormone-binding-globuline (SHBG) levels was evidenced in the paroxetine group (p = 0.01) as a sign of improved insulin sensitivity. There was also a trend for superior efficacy of paroxetine in investigator-rated anxiety and depression. This notion was supported by a trend for superior decrease of serum cortisol levels in the paroxetine group (p = 0.06). CONCLUSION: Paroxetine has a beneficial effect on measures of insulin sensitivity and may improve glycemic control. Larger studies of longer duration are needed to verify the benefits of paroxetine in type 2 diabetes. While waiting for more conclusive evidence it seems sensible to augment standard care of type 2 diabetes with paroxetine even in patients who do not fulfil routine psychiatric criteria for initiation of antidepressant drug treatment. PMID- 12747812 TI - [Noninvasive positive pressure ventilation for acute respiratory failure in chronic obstructive pulmonary disease in a general respiratory ward]. AB - BACKGROUND AND OBJECTIVE: In patients with acute exacerbations of chronic obstructive pulmonary disease (COPD), noninvasive ventilatory support (NPPV) with bilevel positive airway pressure (BiPAP) may improve clinical and physiological parameters. The present study used a randomized, prospective design to evaluate the possible benefits of NPPV plus standard therapy versus standard therapy alone in patients admitted with acute hypercapnic respiratory failure in a respiratory unit of a tertiary hospital. PATIENTS AND METHOD: Forty-one patients were included in the study. Of them, 20 were randomly allocated to receive NPPV with a standard mask connected to a BiPAP ventilatory assist device (Respironics Inc, Murrysville, PA) and 21 to standard therapy. Both groups had similar characteristics upon their admission in the hospital. RESULTS: The use of noninvasive ventilation significantly reduced the respiratory rates and improved the conscious level within the first 2 h (p < 0.001). There were significant differences in PaCO2 and pH (p < 0.05) at 6 h of treatment. The need for intubation was 5% in the NPPV group vs 14% in the control group. The length of hospital stay was significantly shorter in the NPPV group (7 vs 10 days; p < 0.01). Nasal NPPV was well tolerated and complications were uncommon and mild. CONCLUSIONS: Early use of noninvasive ventilation in patients with acute exacerbation of chronic obstructive pulmonary disease leads to a more rapid improvement of physiological variables. Moreover, it is possible to apply this treatment in a general respiratory ward. PMID- 12747811 TI - [Effect of enzyme replacement therapy on lipid profile in patients with Gaucher's disease]. AB - BACKGROUND AND OBJECTIVE: Gaucher's disease (GD) is a lysosomal storage disorder, caused by a deficiency of the acid -glucocerebrosidase enzyme, which results in accumulation of lipids within macrophages. GD patients show decreased plasma total cholesterol (TC), low-density lipoprotein cholesterol (LDL-c) and high density lipoprotein cholesterol (HDL-c) levels, as well as decreased apolipoprotein (apo) A-I and B. Conversely, concentrations of plasma apo E and the chitotriosidase (ChT) activity, a chitinase synthesized by activated macrophages, are increased. Enzyme replacement therapy (ERT) is effective and safe in reversing the clinical manifestations in symptomatic patients. The aim of this study was to evaluate the effect of ERT on the lipid profile of GD patients. PATIENTS AND METHOD: 70 patients, from the Spanish Registry of Gaucher's disease (REEG), were divided into two groups: 54 under ERT, according to stablished criteria, and 16 without ERT. Plasma apolipoprotein, lipoprotein and lipid concentrations, and chitotriosidase activity were analyzed in both groups. Statistical analysis was carried out with the U-Mann Whitney non-parametric test for comparison of data. RESULTS: The group of GD patients under ERT showed significant increases of HDL-c (+38%) and apo A-I (+18%) levels, whereas no changes were observed in LDL-c and apo B levels. Conversely, chitotriosidase activity (-58%), plasma-apo E (-32%) and HDL-apo E (-26%) levels were dramatically reduced after ERT. No significant modifications were observed in the group without ERT. CONCLUSIONS: ERT in GD patients displays significant effects on the concentration of plasma lipoproteins, resulting in a less atherogenic lipid profile and in a reduction of activated macrophages. PMID- 12747813 TI - [Measuring psychosocial stress at work in Spanish hospital's personnel. Psychometric properties of the Spanish version of Effort-Reward Imbalance model]. AB - BACKGROUND AND OBJECTIVE: Two main models are currently used to evaluate the psychosocial factors at work: the Demand-Control (or job strain) model developed by Karasek and the Effort-Reward Imbalance model, developed by Siegrist. A Spanish version of the first model has been validated, yet so far no validated Spanish version of the second model is available. The objective of this study was to explore the psychometric properties of the Spanish version of the Effort Reward Imbalance model in terms of internal consistency, factorial validity, and discriminate validity. SUBJECTS AND METHODS: A cross-sectional study on a representative sample of 298 workers of the Spanish public hospital San Agustin in Asturias was performed. The Spanish version of Effort-Reward Imbalance Questionnaire (23 items) was obtained by a standard forward/backward translation procedure, and the information was gathered by a self-administered application. Exploratory factor analysis were performed to test the dimensional structure of the theoretical model. Cronbach's alpha coefficient was calculated to estimate the internal consistency reliability. Information on discriminate validity is given for sex, age and education. Differences were calculated with the t-test for two independent samples or ANOVA, respectively. RESULTS: Internal consistency was satisfactory for the two scales (reward and intrinsic effort) and Cronbach's Alpha coefficients higher than 0.80 were observed. The internal consistency for the scale of extrinsic effort was lower (alpha = 0.63). A three-factor solution was retained for the factor analysis of reward as expected, and these dimensions were interpreted as a) esteem, b) job promotion and salary and c) job instability. A one-factor solution was retained for the factor analysis of intrinsic effort. The factor analysis of the scale of extrinsic effort did not support the expected one-dimension structure. The analysis of discriminate validity displayed significant associations between measures of Effort-Reward Imbalance and the variables of sex, age and education level. CONCLUSIONS: This study is the first one supporting satisfactory psychometric properties of the Spanish version of the Effort-Reward Imbalance model. However, the factorial validity of the extrinsic effort could be questioned. PMID- 12747815 TI - [Medical specialties in Spain: a window to modernization?]. PMID- 12747814 TI - [Association between non-steroid antiinflammatory drugs intake and decompensation of heart failure]. AB - BACKGROUND AND OBJECTIVE: Our purpose was to estimate the association between the intake of non-steriodal anti-inflammatory drugs (NSAIDs) and hospitalization due to heart failure (HF) and to assess the effect of selective COX-2 inhibitors. PATIENTS AND METHOD: This was a case-control study, including 982 patients with HF, and 788 controls, with a history of HF who were hospitalized in 10 hospitals of the community of Madrid. RESULTS: The use of NSAIDs (other than aspirine at low doses) was associated with an increase in the risk of hospitalization due to HF (OR crude = 1.59; 95% interval confidence, 1.20-2.09; OR adjusted, 1.40; 95% interval confidence, 1.03-1.90). CONCLUSIONS: Intake of NSAIDs is associated with an increased risk of hospitalization due to HF, in susceptible patients. PMID- 12747816 TI - [Facilitation of telephone access to 061 emergency Health Services for deaf people]. PMID- 12747817 TI - [Non-alcoholic steatohepatitis]. PMID- 12747818 TI - [Self limited necrotizing sarcoid granulomatosis]. PMID- 12747819 TI - [Acute abdomen due to pica]. PMID- 12747820 TI - [Fever due to Epstein-Barr virus infection in Crohns disease]. PMID- 12747821 TI - [Usefulness of megestrol acetate for weight gain in neoplastic patients with cachexia]. PMID- 12747823 TI - [Acute hepatitis due to bentazepam]. PMID- 12747824 TI - [Current Nuclear Medicine, the result of a solid and laborious past]. PMID- 12747825 TI - [Neurological explorations in Nuclear Medicine. A Spanish survey]. AB - A representative Nuclear Medicine Service in Spain has two or three gammacameras. It performs neurologic studies one or two days at week wich account 2,1% of total workload. Brain Perfusion SPECT, specially in cognitive disorders, is the most frequent application. Neurooncology has a lower but established relevance. Neuroreceptors imaging are increasing in the last months. Emission tomography is obtained using a double-headed camera fitted with high-resolution parallel hole collimators and a half an hour total acquisition time. Datasets are reconstructed by filtered backprojection with Butterworth or Metz filtres. Images are visually interpreted with comparison to MRI and/or CT findings. PMID- 12747827 TI - TOS motif-mediated raptor binding regulates 4E-BP1 multisite phosphorylation and function. AB - BACKGROUND: The mammalian target of rapamycin, mTOR, is a serine/threonine kinase that controls cell growth and proliferation via the translation regulators eukaryotic initiation factor 4E (eIF4E) binding protein 1 (4E-BP1) and ribosomal protein S6 kinase 1 (S6K1). We recently identified a TOR signaling (TOS) motif in the N terminus of S6K1 and the C terminus of 4E-BP1 and demonstrated that in S6K1, the TOS motif is necessary to facilitate mTOR signaling to phosphorylate and activate S6K1. However, it is unclear how the TOS motif in S6K1 and 4E-BP1 mediates mTOR signaling. RESULTS: Here, we show that a functional TOS motif is required for 4E-BP1 to bind to raptor (a recently identified mTOR-interacting protein), for 4E-BP1 to be efficiently phosphorylated in vitro by the mTOR/raptor complex, and for 4E-BP1 to be phosphorylated in vivo at all identified mTOR regulated sites. mTOR/raptor-regulated phosphorylation is necessary for 4E-BP's efficient release from the translational initiation factor eIF4E. Consistently, overexpression of a mutant of 4E-BP1 containing a single amino acid change in the TOS motif (F114A) reduces cell size, demonstrating that mTOR-dependent regulation of cell growth by 4E-BP1 is dependent on a functional TOS motif. CONCLUSIONS: Our data demonstrate that the TOS motif functions as a docking site for the mTOR/raptor complex, which is required for multisite phosphorylation of 4E-BP1, eIF4E release from 4E-BP1, and cell growth. PMID- 12747828 TI - MicroRNAs and other tiny endogenous RNAs in C. elegans. AB - BACKGROUND: MicroRNAs (miRNAs) are small noncoding RNAs that are processed from hairpin precursor transcripts by Dicer. miRNAs probably inhibit translation of mRNAs via imprecise antisense base-pairing. Small interfering RNAs (siRNAs) are similar in size to miRNAs, but they recognize targets by precise complementarity and elicit RNA-mediated interference (RNAi). We employed cDNA sequencing and comparative genomics to identify additional C. elegans small RNAs with properties similar to miRNAs and siRNAs. RESULTS: We found three broad classes of small RNAs in C. elegans: (1) 21 new miRNA genes (we estimate that C. elegans contains approximately 100 distinct miRNA genes, about 30% of which are conserved in vertebrates; (2), 33 distinct members of a class of tiny noncoding RNA (tncRNA) genes with transcripts that are similar in length to miRNAs (approximately 20-21 nt) and that are in some cases developmentally regulated but are apparently not processed from a miRNA-like hairpin precursor and are not phylogenetically conserved; (3) more than 700 distinct small antisense RNAs, about 20 nt long, that are precisely complementary to protein coding regions of more than 500 different genes and therefore seem to be endogenous siRNAs. CONCLUSIONS: The presence of diverse endogenous siRNAs in normal worms suggests ongoing, genome wide gene silencing by RNAi. miRNAs and tncRNAs are not predicted to form complete Watson-Crick hybrids with any C. elegans RNA target, and so they are likely to regulate the activity of other genes by non-RNAi mechanisms. These results suggest that diverse modes of small RNA-mediated gene regulation are deployed in normal worms. PMID- 12747829 TI - Differential activation of the DNA replication checkpoint contributes to asynchrony of cell division in C. elegans embryos. AB - BACKGROUND: Acquisition of lineage-specific cell cycle duration is a central feature of metazoan development. The mechanisms by which this is achieved during early embryogenesis are poorly understood. In the nematode Caenorhabditis elegans, differential cell cycle duration is apparent starting at the two-cell stage, when the larger anterior blastomere AB divides before the smaller posterior blastomere P(1). How anterior-posterior (A-P) polarity cues control this asynchrony remains to be elucidated. RESULTS: We establish that early C. elegans embryos possess a hitherto unrecognized DNA replication checkpoint that relies on the PI-3-like kinase atl-1 and the kinase chk-1. We demonstrate that preferential activation of this checkpoint in the P(1) blastomere contributes to asynchrony of cell division in two-cell-stage wild-type embryos. Furthermore, we show that preferential checkpoint activation is largely abrogated in embryos that undergo equal first cleavage following inactivation of Galpha signaling. CONCLUSION: Our findings establish that differential checkpoint activation contributes to acquisition of distinct cell cycle duration in two-cell-stage C. elegans embryos and suggest a novel mechanism coupling asymmetric division to acquisition of distinct cell cycle duration during development. PMID- 12747830 TI - Flamingo regulates R8 axon-axon and axon-target interactions in the Drosophila visual system. AB - Photoreceptors (R cells) in the Drosophila retina connect to targets in three distinct layers of the optic lobe of the brain: R1-R6 connect to the lamina, and R7 and R8 connect to distinct layers in the medulla. In each of these layers, R axon termini are arranged in evenly spaced topographic arrays. In a genetic screen for mutants with abnormal R cell connectivity, we recovered mutations in flamingo (fmi). fmi encodes a seven-transmembrane cadherin, previously shown to function in planar cell polarity and in dendritic patterning. Here, we show that fmi has two specific functions in R8 axon targeting: it facilitates competitive interactions between adjacent R8 axons to ensure their correct spacing, and it promotes the formation of stable connections between R8 axons and their target cells in the medulla. The former suggests a general role for Fmi in establishing nonoverlapping dendritic and axonal target fields. The latter, together with the finding that N-Cadherin has an analogous role in R7 axon-target interactions, points to a cadherin-based system for target layer specificity in the Drosophila visual system. PMID- 12747831 TI - Superior underwater vision in a human population of sea gypsies. AB - Humans are poorly adapted for underwater vision. In air, the curved corneal surface accounts for two-thirds of the eye's refractive power, and this is lost when air is replaced by water. Despite this, some tribes of sea gypsies in Southeast Asia live off the sea, and the children collect food from the sea floor without the use of visual aids. This is a remarkable feat when one considers that the human eye is not focused underwater and small objects should remain unresolved. We have measured the visual acuity of children in a sea gypsy population, the Moken, and found that the children see much better underwater than one might expect. Their underwater acuity (6.06 cycles/degree) is more than twice as good as that of European children (2.95 cycles/degree). Our investigations show that the Moken children achieve their superior underwater vision by maximally constricting the pupil (1.96 mm compared to 2.50 mm in European children) and by accommodating to the known limit of human performance (15-16 D). This extreme reaction-which is routine in Moken children-is completely absent in European children. Because they are completely dependent on the sea, the Moken are very likely to derive great benefit from this strategy. PMID- 12747832 TI - PROPORZ1, a putative Arabidopsis transcriptional adaptor protein, mediates auxin and cytokinin signals in the control of cell proliferation. AB - Plants generate cells and organs throughout their life cycle. Plant cell proliferation relates to the activity of dividing meristematic cells, which subsequently differentiate in a position- and lineage-dependent manner. The events underlying the regulation of cell division and further differentiation processes are under tight control of both intrinsic and environmental parameters. Among the intrinsic factors, two groups of phytohormones, auxins and cytokinins, exhibit a combined regulatory impact on cell proliferation, as an important determinant for the totipotency of plant cells. Classical experiments demonstrated that application of both growth regulators in appropriate concentrations promotes callus formation. When the ratio between the phytohormones changes, callus cells acquire the competence to regenerate organs. Typically, a high auxin-to-cytokinin ratio promotes the formation of roots, whereas a low auxin-to-cytokinin ratio results in the regeneration of shoots. Conclusively, the concerted, proportional impact of both phytohormones functions as a determinant of plant cell proliferation; they act as cell cycle-promoting mitogens as well as morphogens that control plant organogenesis. Here, we describe PROPORZ1 (PRZ1), an Arabidopsis gene, essential for the developmental switch from cell proliferation to differentiation in response to variations in auxin and cytokinin concentrations. PRZ1 probably acts as a transcriptional adaptor protein that affects the expression of cell cycle regulators and might, thereby, mediate its effect on the control of cell proliferation. PMID- 12747834 TI - Reconstituted skin from murine embryonic stem cells. AB - Embryonic stem (ES) cell lines can be expanded indefinitely in culture while maintaining their potential to differentiate into any cell type. During embryonic development, the skin forms as a result of reciprocal interactions between mesoderm and ectoderm. Here, we report the in vitro differentiation and enrichment of keratinocytes from murine ES cells seeded on extracellular matrix (ECM) in the presence of Bone Morphogenic Protein-4 (BMP-4) or ascorbate. The enriched preparation of keratinocytes was able to form an epidermal equivalent composed of a stratified epithelium when cultured at the air-liquid interface on a collagen-coated acellular substratum. Interestingly, an underlying cellular compartment that belongs to the fibroblast lineage was systematically formed between the reconstituted epidermis and the inert membrane. The resulting tissue displayed morphological patterns similar to normal embryonic skin, as evidenced by light and transmission electron microscopy. Immunohistochemical studies revealed expression patterns of cytokeratins, basement membrane (BM) proteins and late differentiation markers of epidermis, as well as fibroblast markers, similar to native skin. The results demonstrate the capacity of ES cells to reconstitute in vitro a fully differentiated skin. This ES-derived bioengineered skin provides a powerful tool for studying the molecular mechanisms controlling epidermal and dermal commitments. PMID- 12747833 TI - Arabidopsis HEN1: a genetic link between endogenous miRNA controlling development and siRNA controlling transgene silencing and virus resistance. AB - In animals, double-stranded short interfering RNA (siRNA) and single-stranded microRNA (miRNA) regulate gene expression by targeting homologous mRNA for cleavage or by interfering with their translation, respectively. siRNAs are processed from injected or transgene-derived, long, perfect double-stranded RNA (dsRNA), while miRNAs are processed from short, imperfect dsRNA precursors transcribed from endogenous intergenic regions. In plants, both siRNAs and miRNAs activate cleavage of homologous RNA targets, but little is known about the genes controlling their production or action. The SGS2/SDE1 protein contributes to produce transgene siRNA, while DCL1 and HEN1 contribute to endogenous miRNA accumulation. Here, we show that: i) SGS2, SGS3, AGO1, and HEN1 contribute to produce transgene siRNA involved in sense posttranscriptional gene silencing (S PTGS); ii) HEN1, but not SGS2, SGS3, or AGO1, contributes to the accumulation of the endogenous miR171 miRNA and to the cleavage of Scarecrow target mRNA by miR171; iii) SGS2, SGS3, AGO1, and HEN1 contribute to resistance against cucumber mosaic virus, but not to siRNA and IR-PTGS triggered by hairpin transgenes directly producing perfect dsRNA; and iv) the actions of HEN1 in miRNA/development and siRNA/S-PTGS can be uncoupled by single-point mutations at different positions in the protein. PMID- 12747835 TI - Either part of a Drosophila epsin protein, divided after the ENTH domain, functions in endocytosis of delta in the developing eye. AB - Epsin is part of a protein complex that performs endocytosis in eukaryotes. Drosophila epsin, Liquid facets (Lqf), was identified because it is essential for patterning the eye and other imaginal disc derivatives [2]. Previous work has provided only indirect evidence that Lqf is required for endocytosis in Drosophila [2, 3]. Epsins are modular and have an N-terminal ENTH (epsin N terminal homology) domain that binds PIP(2) at the cell membrane and four different classes of protein-protein interaction motifs. The current model for epsin function in higher eukaryotes is that epsin bridges the cell membrane, a transmembrane protein to be internalized, and the core endocytic complex. Here, we show directly that Drosophila epsin (Lqf) is required for endocytosis. Specifically, we find that Lqf is essential for internalization of the Delta (Dl) transmembrane ligand in the developing eye. Using this endocytic defect in lqf mutants, we develop a transgene rescue assay and perform a structure/function analysis of Lqf. We find that when we divide Lqf into two pieces, an ENTH domain and an ENTH-less protein, each part retains significant ability to function in Dl internalization and eye patterning. These results challenge the model for epsin function that requires an intact protein. PMID- 12747836 TI - A novel domain in AMP-activated protein kinase causes glycogen storage bodies similar to those seen in hereditary cardiac arrhythmias. AB - The AMP-activated protein kinase (AMPK) is an alphabetagamma heterotrimer that is activated by low cellular energy status and affects a switch away from energy requiring processes and toward catabolism. While it is primarily regulated by AMP and ATP, high muscle glycogen has also been shown to repress its activation. Mutations in the gamma2 and gamma3 subunit isoforms lead to arrhythmias associated with abnormal glycogen storage in human heart and elevated glycogen in pig muscle, respectively. A putative glycogen binding domain (GBD) has now been identified in the beta subunits. Coexpression of truncated beta subunits lacking the GBD with alpha and gamma subunits yielded complexes that were active and normally regulated. However, coexpression of alpha and gamma with full-length beta caused accumulation of AMPK in large cytoplasmic inclusions that could be counterstained with anti-glycogen or anti-glycogen synthase antibodies. These inclusions were not affected by mutations that increased or abolished the kinase activity and were not observed by using truncated beta subunits lacking the GBD. Our results suggest that the GBD binds glycogen and can lead to abnormal glycogen containing inclusions when the kinase is overexpressed. These may be related to the abnormal glycogen storage bodies seen in heart disease patients with gamma2 mutations. PMID- 12747837 TI - AMPK beta subunit targets metabolic stress sensing to glycogen. AB - AMP-activated protein kinase (AMPK) is a multisubstrate enzyme activated by increases in AMP during metabolic stress caused by exercise, hypoxia, lack of cell nutrients, as well as hormones, including adiponectin and leptin. Furthermore, metformin and rosiglitazone, frontline drugs used for the treatment of type II diabetes, activate AMPK. Mammalian AMPK is an alphabetagamma heterotrimer with multiple isoforms of each subunit comprising alpha1, alpha2, beta1, beta2, gamma1, gamma2, and gamma3, which have varying tissue and subcellular expression. Mutations in the AMPK gamma subunit cause glycogen storage disease in humans, but the molecular relationship between glycogen and the AMPK/Snf1p kinase subfamily has not been apparent. We show that the AMPK beta subunit contains a functional glycogen binding domain (beta-GBD) that is most closely related to isoamylase domains found in glycogen and starch branching enzymes. Mutation of key glycogen binding residues, predicted by molecular modeling, completely abolished beta-GBD binding to glycogen. AMPK binds to glycogen but retains full activity. Overexpressed AMPK beta1 localized to specific mammalian subcellular structures that corresponded with the expression pattern of glycogen phosphorylase. Glycogen binding provides an architectural link between AMPK and a major cellular energy store and juxtaposes AMPK to glycogen bound phosphatases. PMID- 12747838 TI - A new mode of Ca2+ signaling by G protein-coupled receptors: gating of IP3 receptor Ca2+ release channels by Gbetagamma. AB - The most common form of Ca(2+) signaling by Gq-coupled receptors entails activation of PLCbeta2 by Galphaq to generate IP(3) and evoke Ca(2+) release from the ER. Another form of Ca(2+) signaling by G protein-coupled receptors involves activation of Gi to release Gbetagamma, which activates PLCbeta1. Whether Gbetagamma has additional roles in Ca(2+) signaling is unknown. Introduction of Gbetagamma into cells activated Ca(2+) release from the IP(3) Ca(2+) pool and Ca(2) oscillations. This can be due to activation of PLCbeta1 or direct activation of the IP(3)R by Gbetagamma. We report here that Gbetagamma potently activates the IP(3) receptor. Thus, Gbetagamma-triggered [Ca(2+)](i) oscillations are not affected by inhibition of PLCbeta. Coimmunoprecipitation and competition experiments with Gbetagamma scavengers suggest binding of Gbetagamma to IP(3) receptors. Furthermore, Gbetagamma inhibited IP(3) binding to IP(3) receptors. Notably, Gbetagamma activated single IP(3)R channels in native ER as effectively as IP(3). The physiological significance of this form of signaling is demonstrated by the reciprocal sensitivity of Ca(2+) signals evoked by Gi- and Gq coupled receptors to Gbetagamma scavenging and PLCbeta inhibition. We propose that gating of IP(3)R by Gbetagamma is a new mode of Ca(2+) signaling with particular significance for Gi-coupled receptors. PMID- 12747839 TI - Selective activation of thrombin is a critical determinant for vertebrate lens regeneration. AB - The regeneration of structures in adult animals depends on a mechanism for coupling the acute response to tissue injury or removal with the local activation of plasticity in residual differentiated cells or stem cells. Many potentially relevant signals are generated after injury, and the nature of this mechanism has not been elucidated for any instance of regeneration. Lens regeneration in adult vertebrates always occurs at the pupillary margin of the dorsal iris, where pigmented epithelial cells (PEC) reenter the cell cycle and transdifferentiate into the lens, but the basis of this striking preference for the dorsal margin over the ventral is unknown. In this study, we report that a critical early event after lentectomy in the newt is the transient and selective activation of thrombin at the dorsal margin. The thrombin activity was blocked with two different irreversible inhibitors and was shown to be strictly required for cell cycle reentry at this location. The axolotl, a related urodele species, can regenerate its limb, but not its lens, and thrombin is activated in the former context, but not the latter. Our results indicate that selective activation of thrombin is the pivotal signal linking tissue injury to the initiation of vertebrate regeneration. PMID- 12747840 TI - Analysis of Myc bound loci identified by CpG island arrays shows that Max is essential for Myc-dependent repression. AB - The c-myc proto-oncogene encodes a transcription factor, c-Myc, which is deregulated and/or overexpressed in many human cancers. Despite c-Myc's importance, the identity of Myc-regulated genes and the mechanism by which Myc regulates these genes remain unclear. By combining chromatin immunoprecipitation with CpG island arrays, we identified 177 human genomic loci that are bound by Myc in vivo. Analyzing a cohort of known and novel Myc target genes showed that Myc-associated protein X, Max, also bound to these regulatory regions. Indeed, Max is bound to these loci in the presence or absence of Myc. The Myc:Max interaction is essential for Myc-dependent transcriptional activation; however, we show that Max bound targets also include Myc-repressed genes. Moreover, we show that the interaction between Myc and Max is essential for gene repression to occur. Taken together, the identification and analysis of Myc bound target genes supports a model whereby Max plays an essential and universal role in the mechanism of Myc-dependent transcriptional regulation. PMID- 12747841 TI - Europe uneasy on stem cell developments. AB - A first round rejection of human stem cell research by the European Parliament has given supporters the jitters in member states keen to pursue the potential offered by the new field, shown by a recent landmark appeal court decision on stem cell therapy by a team of Britain's top judges. PMID- 12747842 TI - Picture this. AB - Some human populations in south-east Asia have foraged for a living in shallow coastal waters for generations. Could their eyes have adapted to these challenging conditions? PMID- 12747843 TI - Yuh-Nung Jan. PMID- 12747845 TI - Protein disulphide isomerase. PMID- 12747846 TI - The neural crest. PMID- 12747847 TI - Chromosome segregation: clamping down on deviant orientations. AB - Chromosome segregation depends on proper orientation of sister kinetochores. The protein Csm1 is required for mono-orientation of sister kinetochores at meiosis I in budding yeast. Surprisingly, its homologue in fission yeast appears instead of clamp micro-tubule binding sites together on single mitotic kinetochores so that they all face one spindle pole. PMID- 12747848 TI - Patterning: JAK-STAT signalling in the Drosophila follicular epithelium. AB - During Drosophila oogenesis the follicular epithelium becomes subdivided into distinct cell populations. New reports have established that the Janus kinase (JAK) signalling pathway plays an important role in this process. PMID- 12747849 TI - ER export: call 14-3-3. AB - Forward transport of proteins from the ER to the plasma membrane requires escape from the ER's retention machinery. Recent studies suggest that 14-3-3 proteins may mediate ER export of potassium channels destined for the plasma membrane by interfering with dibasic-motif-mediated retention. PMID- 12747850 TI - Stereoscopic vision: solving the correspondence problem. AB - Neurons in early visual areas respond to horizontal disparity in images that do not give rise to stereopsis. False binocular matches, however, are discarded at the apex of the visual pathway: the activity of neurons in the primate inferior temporal cortex correlates directly with conscious depth perception. PMID- 12747851 TI - Secretion: kiss and run caught on film. AB - Recent results have provided graphic support for the hypothesis that vesicle secretion involves a 'kiss-and-run' mechanism. Evanescent field microscopy has shown that, during exocytosis, intravesicular markers escape without collapse of the vesicular membrane into the surface membrane and that the empty vesicle is immediately retrieved back into the cell. PMID- 12747853 TI - Insulin secretion: fatty acid signalling via serpentine receptors. AB - Insulin secretion is thought principally to be regulated by blood glucose concentration. Three recent studies emphasise the additional importance of fatty acids as regulators of insulin secretion, and demonstrate the involvement of a novel G protein-coupled receptor. PMID- 12747852 TI - Plant pathology: monitoring a pathogen-targeted host protein. AB - A plant protein RIN4 is targeted and modified by bacterial pathogens as part of the disease process. At least two host resistance proteins monitor this pathogen interference and trigger the plant's defence responses. PMID- 12747854 TI - Auditory pathways: are 'what' and 'where' appropriate? AB - New evidence confirms that the auditory system encompasses temporal, parietal and frontal brain regions, some of which partly overlap with the visual system. But common assumptions about the functional homologies between sensory systems may be misleading. PMID- 12747855 TI - Control of vesicular trafficking by Rho GTPases. AB - Although vesicular trafficking is essential for a large variety of cellular processes, the regulation of vesicular trafficking is still poorly understood. Members of the Rho family of small GTPases have recently emerged as important control elements of many stages of vesicular trafficking, providing new insight into the regulation of these events. We will discuss the diverse roles played by Rho proteins in membrane trafficking and focus on the biological implications of these functions. PMID- 12747857 TI - Recombinant (2-->3)-alpha-sialyltransferase immobilized on nickel-agarose for preparative synthesis of sialyl Lewis(x) and Lewis(a) precursor oligosaccharides. AB - The specificity of recombinant (2-->3)-alpha-sialyltransferase (ST3Gal-III), expressed in baculovirus-infected insect cells, has been determined with various oligosaccharide acceptors and sugar-nucleotide donors using a fluorescence based assay. Recombinant ST3Gal-III tagged with a polyhistidine tail was immobilized on Ni(2+)-NTA-Agarose as an active enzyme for use in the synthesis of three sialylated oligosaccharides: (i) the divalent molecule [alpha-Neu5Ac-(2-->3)-D Galp-(1-->4)-beta-D-GlcpNAc-O-CH(2)](2)-C-(CH(2)OBn)(2) (12); (ii) the dansylated derivative, alpha-Neu5Ac-(2-->3)-D-Galp-(1-->3)-beta-D-GlcpNAc-O-(CH(2))(6)-NH dansyl and; (iii) the tetrasacharide alpha-Neu5Ac-(2-->3)-beta-D-Galp-(1-->4) beta-D-GlcpNAc-(1-->2)-alpha-D-Manp-O-CH(3). Compound 12 was itself prepared from the divalent N-acetyllactosamine molecule built on pentaerythritol by a chemo enzymatic route. PMID- 12747858 TI - Chemo-enzymatic synthesis of a divalent sialyl Lewis(x) ligand with restricted flexibility. AB - To study the influence of the entropic factor in cluster cooperative effects, a divalent sialyl Lewis(x) ligand with restricted flexilbility was chemo enzymatically synthesized. First, a cyclized precursor with both glucosamine residues bridged together by a succinyl group was readily obtained in 42% yield by treatment of 2,2-bis(benzyloxymethyl)-1,3-bis(3,4,6-tri-O-acetyl-2-amino-2 deoxy-beta-D-glucopyranosyloxy)-propane with succinyl chloride. After deacetylation, this precursor was subjected to stepwise enzymatic elongation utilizing successively, soluble galactosyltransferase, then recombinant sialyltransferase and fucosyltransferase; the latter enzymes immobilized on Ni(2+)-Agarose, to afford, after debenzylation, a divalent sialyl Lewis(x) ligand of restricted flexibility, in 45% overall yield. Following the same enzymatic sequence, a totally flexible ligand, required as a reference compound for evaluation of inhibitory activity toward selectins, was also prepared from 2,2(bis-benzyloxymethyl)-1,3-bis(2-acetamido-2-deoxy-beta-D-glucopyranosyloxy) propane, as well as both related divalent Lewis(x) molecules lacking the sialic acids, the rigid one and the flexible one. PMID- 12747859 TI - A new mannoheptaose containing alpha and beta-(1-->2) linkages isolated from the mannan of Torulaspora delbrueckii: ELISA inhibition studies. AB - Torulaspora delbrueckii starin IFO 0955 was examined with respect to its structural and serological properties of the cell wall mannan (Td-0955-M). Td 0955-M revealed significant reactivities with sera from a commercially available factor serum kit (Candida Check) in ELISA. Td-0955-M was investigated for its chemical structure by acetolysis under conventional and mild conditions. NMR and GC techniques were used as analytical techniques. The mannooligosaccharide fractions eluted from a Bio-Gel P-2 column were found to consist of Man(alpha1 2)Man, M2, Man(alpha1-2)Man(alpha1-2)Man and Man(beta1-2)Man(alpha1-2)Man, M3, Man(alpha1-2)Man(beta1-2)Man(beta1-2)Man(alpha1-2)Man, M5, and a new mannoheptaose, which possesses the structure, Man(alpha1-2)Man(beta1-2)Man(beta1 2)Man(beta1-2)Man(beta1-2)Man(alpha1-2)Man, M7. The results of the inhibition ELISA showed that the M7 oligosaccharide significantly inhibited the reactivities in the Td-0955-M-factor serum systems. PMID- 12747860 TI - Dextran molecular size and degree of branching as a function of sucrose concentration, pH, and temperature of reaction of Leuconostoc mesenteroides B 512FMCM dextransucrase. AB - Reactions of Leuconostoc mesenteroides B-512FMCM dextransucrase with increasing concentrations of sucrose, from 0.1 to 4.0 M, gave a decreasing amount of high molecular weight dextran (HMWD) (>10(6) Da) with a concomitant increase in low molecular weight dextran (LMWD) (<10(5) Da). At 0.1 M sucrose, pH 5.5, and 28 degrees C, 99.8% of the dextran had a MW>10(6) Da and at 4.0 M sucrose, 69.9% had a MW<10(5) Da and 30.1% had a MW>10(6) Da, giving a bimodal distribution. The degree of branching increased from 5% for 0.1 M sucrose to 16.6% for 4.0 M sucrose. The temperature had very little effect on the size of the dextran, which was >10(6) Da, but it had a significant effect on the degree of branching, which was 4.8% at 4 degrees C and increased to 14.7% at 45 degrees C. Both the molecular weight (MW) and the degree of branching were not significantly affected by different pH values between 4.5 and 6.0. PMID- 12747861 TI - Structural and serological studies of the O-antigen of Proteus mirabilis O-9. AB - The following structure of the O-polysaccharide (O-antigen) of the lipopolysaccharide of Proteus mirabilis O-9 was determined by NMR spectroscopy, including 2D 1H,(1)H COSY, TOCSY, ROESY, and 1H,(13)C HMQC experiments, along with chemical methods: [chemical structure: see text] where the degree of O acetylation is approximately 70%. Immunochemical studies using rabbit polyclonal anti-Proteus mirabilis O-9 serum showed the importance of the O-acetyl groups in manifesting the serological specificity of the O-9 antigen. Anti-P. mirabilis O-9 cross-reacted with the lipopolysaccharides (LPS) of P. vulgaris O-25 and Proteus penneri 14, which could be accounted for by a structural similarity of their O polysaccharides. PMID- 12747862 TI - Structure of the xyloglucan produced by suspension-cultured tomato cells. AB - The xyloglucan secreted by suspension-cultured tomato (Lycopersicon esculentum) cells was structurally characterized by analysis of the oligosaccharides generated by treating the polysaccharide with a xyloglucan-specific endoglucanase (XEG). These oligosaccharide subunits were chemically reduced to form the corresponding oligoglycosyl alditols, which were isolated by high-performance liquid chromatography (HPLC). Thirteen of the oligoglycosyl alditols were structurally characterized by a combination of matrix-assisted laser-desorption ionization mass spectrometry and two-dimensional nuclear magnetic resonance (NMR) spectroscopy. Nine of the oligoglycosyl alditols (GXGGol, XXGGol, GSGGol, XSGGol, LXGGol, XTGGol, LSGGol, LLGGol, and LTGGol, [see, Fry, S.C.; York, W.S., et al., Physiologia Plantarum 1993, 89, 1-3, for this nomenclature]) are derived from oligosaccharide subunits that have a cellotetraose backbone. Very small amounts of oligoglycosyl alditols (XGGol, XGGXXGGol, XXGGXGGol, and XGGXSGGol) derived from oligosaccharide subunits that have a cellotriose or celloheptaose backbone were also purified and characterized. The results demonstrate that the xyloglucan secreted by suspension-cultured tomato cells is very complex and is composed predominantly of 'XXGG-type' subunits with a cellotetraose backbone. The rigorous characterization of the oligoglycosyl alditols and assignment of their 1H and 13C NMR spectra constitute a robust data set that can be used as the basis for rapid and accurate structural profiling of xyloglucans produced by Solanaceous plant species and the characterization of enzymes involved in the synthesis, modification, and breakdown of these polysaccharides. PMID- 12747863 TI - Alkaline degradation kinetics and CE-separation of cello- and xylooligomers. Part I. AB - The degradation kinetics of cello- and xylooligomers under alkaline conditions has been studied, and kinetic order, kinetic rate constants as well as activation parameters (E(A), DeltaE(#), DeltaS(#)) for the different oligomers have been determined. The results were corroborated by a mathematical model of the degradation kinetics. A reliable and convenient method for the separation and simultaneous quantification of cello- and xylooligomers, based on capillary electrophoresis (CE) with pre-column derivatization, has been established. p Aminobenzonitrile was used as the UV tag, and 550 mM borate buffer containing hexadimethrine bromide was employed as the running electrolyte. PMID- 12747864 TI - Electrochemical study of the complexes of aspartame with Cu(II), Ni(II) and Zn(II) ions in the aqueous medium. AB - The voltammetric behaviours of aspartame in the presence of some metal ions (Cu(II), Ni(II), Zn(II)) were investigated. In the presence of aspartame, copper ions reduced at two stages with quasi-reversible one-electron and, with increasing the aspartame (L) concentration, Cu(II)L(2) complex reduces at one stage with irreversible two-electron reaction (-0.322 V). Zn(II)-aspartame complex (logbeta=3.70) was recognized by a cathodic peak at -1.320 V. Ni(II) aspartame complex (logbeta=6.52) is reduced at the more positive potential (-0.87 V) than that of the hydrated Ni(II) ions (-1.088 V). In the case of the reduction of Ni(II) ions, aspartame serves as a catalyst. From electronic spectra data of the complexes, their stoichiometries of 1:2 (metal-ligand) in aqueous medium are determined. The greatness of these logarithmic values is agreement with Irwing Williams series (NiZn). PMID- 12747865 TI - Structural analysis of the lipooligosaccharide from the commensal Haemophilus somnus strain 1P. AB - The structure of the lipooligosaccharide (LOS) from the commensal Haemophilus somnus strain 1P was elucidated. The structure of the O-deacylated LOS was established by monosaccharide analysis, NMR spectroscopy and mass spectrometry. The following structure for the O-deacylated LOS was determined on the basis of the combined data from these experiments. [chemical structure: see text] In the structure Kdo is 3-deoxy-D-manno-octulosonic acid, Hep is L-glycero-D-manno heptose and lipid A-OH refers to O-deacylated Lipid A. The elucidation of this structure has increased our understanding of the relationship between the variability in LOS structure and the pathogenic potential of this organism. Specifically, the inability of this commensal strain to sialylate its LOS suggests that LOS sialylation could be a crucial virulence factor for H. somnus. PMID- 12747866 TI - Two different molecular conformations found in chitosan type II salts. AB - The type II structure of chitosan acidic salts prepared from crab tendon in solid state was studied using an X-ray fiber diffraction technique together with the linked-atom least-squares (LALS) technique. The cylindrical Patterson method was applied to confirm the molecular conformation of the chitosan. It was shown that there are two different helical conformations for type II salts. One is the relaxed twofold helix having a tetrasaccharide as an asymmetric unit as found in chitosan.HCl salt, which was previously reported as a conformation of chitosan.HCOOH salt. The other is the fourfold helix having a disaccharide as an asymmetric unit newly found in chitosan.HI salt. PMID- 12747867 TI - Structural elucidation and antitumor activity of a fructan from Cyathula officinalis Kuan. AB - A fructan named CoPS3 was isolated from Cyathula officinalis Kuan. The structure of CoPS3 was determined by methylation, by the reductive-cleavage method combined with GC-MS analysis, and both 1D and 2D 1H and 13C NMR spectroscopy. These results show that CoPS3 is a graminans-type fructan that is comprised of a beta-D fructofuranosyl backbone having residues linked (2-->1)- and (2-->6) with branches and an alpha-D-glucopyranose residue on the nonreducing end of the fructan chain. Each branch is terminated by a beta-D-Fruf residue. Bioassay showed that it could inhibit growth of Lewis pulmonary carcinoma implanted in mice. PMID- 12747869 TI - Structural determination of the O-specific chain of the lipopolysaccharide from the mushrooms pathogenic bacterium Pseudomonas tolaasii. AB - The complete structure of the O-specific polysaccharide of the lipopolysaccharide isolated from the cultivated mushrooms pathogen Pseudomonas tolaasii is described. The structural determination, achieved by chemical and spectroscopical analyses, indicates a novel tetrasaccharide repeating unit built up of two units of 2-acetamido-2,6-di-deoxy-glucopyranose (Quinovosamine, QuipNAc) and two units of 2-acetamido-2-deoxy-gulopyranuronamide (GulpNAcAN), one of which is acetylated at C-3 position: PMID- 12747868 TI - Efficient nitrogen alkylation with carbohydrates. AB - Efficient nitrogen alkylation of various primary and secondary amines, including cyclic, heterocyclic and alkaloid type amines, with a sugar oxetane 3,5-anhydro 1,2-O-cyclohexylidene-alpha-D-xylofuranose is described. As a result, 5-amino-5 deoxy derivatives of xylofuranose were obtained in good yields. PMID- 12747871 TI - Agents of mass destruction found in USA. PMID- 12747872 TI - DNA microarrays in breast cancer: the promise of personalised medicine. PMID- 12747873 TI - Enfuvirtide, a new drug for HIV infection. PMID- 12747874 TI - The G8 and access to medicines: no more broken promises. PMID- 12747875 TI - The dangerous rise of American exceptionalism. PMID- 12747876 TI - New generation antipsychotics versus low-potency conventional antipsychotics: a systematic review and meta-analysis. AB - BACKGROUND: The clearest advantage of new generation, atypical antipsychotics is a reduced risk of extrapyramidal side-effects (EPS), compared with conventional compounds. These findings might have been biased by the use of the high-potency antipsychotic haloperidol as a comparator in most of the trials. We aimed to establish whether the new drugs induce fewer EPS than low-potency conventional antipsychotics. METHODS: We did a meta-analysis of all randomised controlled trials in which new generation antipsychotics had been compared with low-potency (equivalent or less potent than chlorpromazine) conventional drugs. We included studies that met quality criteria A or B in the Cochrane Collaboration Handbook, and assessed quality with the Jadad scale. The primary outcome of interest was the number of patients who had at least one EPS. We used risk differences and 95% CIs as measures of effect size. FINDINGS: We identified 31 studies with a total of 2320 participants. Of the new generation drugs, only clozapine was associated with significantly fewer EPS (RD=-0.15, 95% CI -0.26 to -0.4, p=0.008) and higher efficacy than low-potency conventional drugs. Reduced frequency of EPS seen with olanzapine was of borderline significance (-0.15, -0.31 to -0.01, p=0.07). Only one inconclusive trial of amisulpride, quetiapine, and risperidone and no investigations of ziprasidone and sertindole were identified, but some evidence indicates that zotepine and remoxipride do not lead to fewer EPS than low-potency antipsychotics. Mean doses less than 600 mg/day of chlorpromazine or its equivalent had no higher risk of EPS than new generation drugs. As a group, new generation drugs were moderately more efficacious than low-potency antipsychotics, largely irrespective of the comparator doses used. INTERPRETATION: Optimum doses of low-potency conventional antipsychotics might not induce more EPS than new generation drugs. Potential advantages in efficacy of the new generation drugs should be a factor in clinical treatment decisions to use these rather than conventional drugs. PMID- 12747877 TI - Diagnosis of viral disease. PMID- 12747878 TI - Gene expression predictors of breast cancer outcomes. AB - BACKGROUND: Correlation of risk factors with genomic data promises to provide specific treatment for individual patients, and needs interpretation of complex, multivariate patterns in gene expression data, as well as assessment of their ability to improve clinical predictions. We aimed to predict nodal metastatic states and relapse for breast cancer patients. METHODS: We analysed DNA microarray data from samples of primary breast tumours, using non-linear statistical analyses to assess multiple patterns of interactions of groups of genes that have predictive value for the individual patient, with respect to lymph node metastasis and cancer recurrence. FINDINGS: We identified aggregate patterns of gene expression (metagenes) that associate with lymph node status and recurrence, and that are capable of predicting outcomes in individual patients with about 90% accuracy. The metagenes defined distinct groups of genes, suggesting different biological processes underlying these two characteristics of breast cancer. Initial external validation came from similarly accurate predictions of nodal status of a small sample in a distinct population. INTERPRETATION: Multiple aggregate measures of profiles of gene expression define valuable predictive associations with lymph node metastasis and disease recurrence for individual patients. Gene expression data have the potential to aid accurate, individualised, prognosis. Importantly, these data are assessed in terms of precise numerical predictions, with ranges of probabilities of outcome. Precise and statistically valid assessments of risks specific for patients, will ultimately be of most value to clinicians faced with treatment decisions. PMID- 12747879 TI - Effectiveness and safety of ICL670 in iron-loaded patients with thalassaemia: a randomised, double-blind, placebo-controlled, dose-escalation trial. AB - BACKGROUND: Transfusional iron overload is a potentially fatal complication of the treatment of thalassaemia. We aimed to investigate short-term efficacy, pharmacokinetic/pharma- codynamic (PK/PD) relations, and safety of ICL670, a novel, tridentate, orally active iron chelator. METHODS: We enrolled 24 patients and divided them into three cohorts consisting of a minimum of seven individuals. Patients were admitted to a metabolic unit and consumed a diet with a defined content of iron. Two patients in each cohort were randomly allocated placebo. Five or more patients received one daily dose of ICL670 at 10, 20, or 40 mg x kg( 1) x day(-1), from day 1 to 12. Net iron excretion (NIE) was measured between days 1 and 12. Primary objectives included assessment of safety and tolerability (measured by adverse events and clinical laboratory monitoring), pharmacokinetics (measured as drug and drug-iron complex), and cumulative net iron excretion (measured by faecal and urine output minus food input). Analysis was for efficacy. FINDINGS: ICL670 was absorbed promptly and was detectable in the blood for 24 h. Exposure (area under the curve of plasma concentration) to ICL670 at pharmacokinetic steady state was proportional to dose. All three doses resulted in positive NIE. The NIE achieved at 20mg x kg(-1) day(-1) would prevent net iron accumulation in most patients transfused with 12-15 mL packed red-blood-cells kg( 1) month(-1), equivalent to 0.3-0.5 mg iron kg(-1) x day(-1). A linear relation (PK/PD) was recorded between exposure to ICL670 and total iron excretion, by contrast with placebo (r2=0.54, p<0.0001). Skin rashes were noted in four patients treated at 20 and 40 mg x kg(-1) x day(-1), and one patient also developed grade 2 transaminitis. INTERPRETATION: ICL670 given once daily at 20 mg/kg seems to be an effective orally active iron chelator and is reasonably well tolerated. Long-term studies are now necessary to establish the practical contribution of this drug. PMID- 12747880 TI - Effect of spiritual well-being on end-of-life despair in terminally-ill cancer patients. AB - BACKGROUND: The importance of spirituality in coping with a terminal illness is becoming increasingly recognised. We aimed to assess the relation between spiritual well-being, depression, and end-of-life despair in terminally-ill cancer patients. METHODS: 160 patients in a palliative care hospital with a life expectancy of less than 3 months were interviewed with a series of standardised instruments, including the functional assessment of chronic illness therapy spiritual well-being scale, the Hamilton depression rating scale, the Beck hopelessness scale, and the schedule of attitudes toward hastened death. Suicidal ideation was based on responses to the Hamilton depression rating scale. FINDINGS: Significant correlations were seen between spiritual well-being and desire for hastened death (r=-0.51), hopelessness (r=-0.68), and suicidal ideation (r=-0.41). Results of multiple regression analyses showed that spiritual well-being was the strongest predictor of each outcome variable and provided a unique significant contribution beyond that of depression and relevant covariates. Additionally, depression was highly correlated with desire for hastened death in participants low in spiritual well-being (r=0.40, p<0.0001) but not in those high in spiritual well-being (r=0.20, p=0.06). INTERPRETATION: Spiritual well-being offers some protection against end-of-life despair in those for whom death is imminent. Our findings have important implications for palliative care practice. Controlled research assessing the effect of spirituality-based interventions is needed to establish what methods can help engender a sense of peace and meaning. PMID- 12747881 TI - Immune tolerance after long-term enzyme-replacement therapy among patients who have mucopolysaccharidosis I. AB - BACKGROUND: Enzyme-replacement therapy has been assessed as a treatment for patients who have mucopolysaccharidosis I (alpha-L-iduronidase deficiency). We aimed to investigate the humoral immune response to recombinant human alpha-L iduronidase among these patients. METHODS: We characterised the antibody titres and specific linear sequence epitope reactivity of serum antibodies to alpha-L iduronidase for ten patients with mucopolysaccharidosis I, at the start of treatment and after 6, 12, 26, 52, and 104 weeks. We compared the values for patients' samples with those for samples from normal human controls. FINDINGS: Before enzyme-replacement therapy, all patients had low serum antibody titres to recombinant human alpha-L-iduronidase that were within the control range. Five of the ten patients produced higher-than-normal titres of antibody to the replacement protein during the treatment course (serum antibody titres 130000 500000 and high-affinity epitope reactivity). However, by week 26, antibody reactivity was reduced, and by week 104 all patients had low antibody titres and only low-affinity epitope reactivity. Patients who had mucopolysaccharidosis I with antibody titres within the normal range at 6-12 weeks did not subsequently develop immune responses. INTERPRETATION: After 2 years of treatment, patients who initially had an immune reaction developed immune tolerance to alpha-L iduronidase. This finding has positive implications for long-term enzyme replacement therapy in patients who have mucopolysaccharidosis I. PMID- 12747882 TI - Neonatal epileptic encephalopathy. PMID- 12747883 TI - Development of a standard treatment protocol for severe acute respiratory syndrome. AB - A series of 31 patients with probable SARS, diagnosed from WHO criteria, were treated according to a treatment protocol consisting of antibacterials and a combination of ribavirin and methylprednisolone. Through experience with the first 11 patients, we were able to finalise standard dose regimens, including pulsed methylprednisolone. One patient recovered on antibacterial treatment alone, 17 showed rapid and sustained responses, and 13 achieved improvement with step-up or pulsed methylprednisolone. Four patients required short periods of non invasive ventilation. No patient required intubation or mechanical ventilation. There was no mortality or treatment morbidity in this series. PMID- 12747884 TI - Acquired mutations in GATA1 in neonates with Down's syndrome with transient myeloid disorder. AB - Transient myeloid disorder is a unique self-regressing neoplasia specific to Down's syndrome. The transcription factor GATA1 is needed for normal growth and maturation of erythroid cells and megakaryocytes. Mutations in GATA1 have been reported in acute megakaryoblastic leukaemia in Down's syndrome. We aimed to investigate changes in GATA1 in patients with Down's syndrome and either transient myeloid disorder (n=10) or acute megakaryoblastic leukaemia (n=6). We recorded mutations eliminating exon 2 from GATA1 in all patients with transient myeloid disorder (age 0-24 days) and in all with acute megakaryoblastic leukaemia (age 14-38 months). The range of mutations did not differ between patients with each disorder. Patients with transient myeloid disorder with mutations in GATA1 can regress spontaneously to complete remission, and mutations do not necessarily predict later acute megakaryoblastic leukaemia. PMID- 12747885 TI - US cancer group calls for centralised review of clinical trials. PMID- 12747886 TI - A look at slowing progression of myopia. PMID- 12747887 TI - Canadians still stung by WHO's SARS travel advisory. PMID- 12747888 TI - Europe contemplates US-style disease-control centre. PMID- 12747889 TI - US smallpox vaccine programme stalls as volunteers balk. PMID- 12747891 TI - New HIV vaccine shows promising results in preclinical studies. PMID- 12747892 TI - UK government faces crunch vote on foundation hospitals. PMID- 12747893 TI - Essential hypertension. AB - Hypertension is a frequent, chronic, age-related disorder, which often entails debilitating cardiovascular and renal complications. Blood pressure is usually noted in combination with other cardiovascular risk factors. Diagnosis of hypertension increasingly relies on automated techniques of blood pressure measurement. The pathophysiology of essential hypertension depends on the primary or secondary inability of the kidney to excrete sodium at a normal blood pressure. The central nervous system, endocrine factors, the large arteries, and the microcirculation also have roles in the disorder. Although monogenic forms of blood pressure dysregulation exist, hypertension mostly arises as a complex quantitative trait that is affected by varying combinations of genetic and environmental factors. Non-pharmacological strategies can reduce blood pressure. Antihypertensive drug treatment diminishes the complications of hypertension. The concept that a few major genes will provide the final clue to the pathogenesis of essential hypertension is an oversimplification that contradicts the heterogeneous nature of this disorder. Further integration of genetic, molecular, clinical, and epidemiological research could disclose subsets of patients in whom specific combinations of genetic and environmental factors raise blood pressure, and might lead to more individualised treatment. PMID- 12747894 TI - Homozygous familial hypercholesterolaemia in identical twins. PMID- 12747895 TI - Huntingtin aggregation and toxicity in Huntington's disease. AB - CONTEXT: Huntington's disease is a late onset neurodegenerative disorder for which the mutation is a CAG/polyglutamine (polyQ) repeat expansion in the gene encoding the huntingtin protein. The disease is one of nine inherited neurodegenerative disorders that are caused by this type of mutation, and which include dentatorubral pallidoluysian atrophy, spinal and bulbar muscular atrophy, and the spinocerebellar ataxias 1, 2, 3, 6, 7, and 17. The mutant proteins are unrelated except for the polyQ tract, and aggregated polyQ is a major component of the proteinaceous deposits that are found in patients' brains for all of these diseases. STARTING POINT: Since the discovery of polyQ aggregates, the proposed role of the aggregation process has ranged from being central to disease pathogenesis, to a benign epiphenomenon, or even to being neuroprotective. Attempts to correlate the presence of aggregates with the onset of phenotype have been complicated by the difficulties in detecting and quantifying small aggregated forms of polyQ, and because all possible structural conformers of the repeat are present in the system under analysis. A paper by W Yang and colleagues (Hum Mol Genet 2002; 11: 2905-17) circumvents these limitations and demonstrates that preformed polyQ aggregates are highly toxic when directed to the cell nucleus. Consistent with aggregate toxicity, pharmacological intervention aimed at inhibiting aggregate formation has recently shown beneficial effects in a mouse model of Huntington's disease (I Sanchez and colleagues, Nature 2003; 421: 373-79). WHERE NEXT: The demonstration that polyQ aggregates are toxic is important because it further validates polyQ aggregation as a therapeutic target. To exploit this finding fully, greater understanding of the formation and structure of polyQ aggregates is needed. However, even without this knowledge, establishing high-throughput screens to identify aggregation inhibitors has been straightforward, and early in-vivo experiments that target aggregation have been promising. As the molecular events that contribute to the early stages of the pathogenesis of Huntington's disease are uncovered, such events will be developed as therapeutic targets. The inhibition of huntingtin aggregation should be a major focus in this effort and the practicalities of this approach are likely to unfold over the next 5-10 years. PMID- 12747896 TI - Keeping an eye on the global traffic in human organs. PMID- 12747897 TI - SARS exposed, pandemic influenza lurks. PMID- 12747898 TI - A fair hearing for the Medical Research Council. PMID- 12747899 TI - A fair hearing for the Medical Research Council. PMID- 12747900 TI - Combined salmeterol and fluticasone for COPD. PMID- 12747901 TI - Combined salmeterol and fluticasone for COPD. PMID- 12747902 TI - Combined salmeterol and fluticasone for COPD. PMID- 12747903 TI - Combined salmeterol and fluticasone for COPD. PMID- 12747904 TI - Combined salmeterol and fluticasone for COPD. PMID- 12747907 TI - WHO to reaffirm health for all. PMID- 12747905 TI - Combined salmeterol and fluticasone for COPD. PMID- 12747908 TI - Low back pain in rural Tibet. PMID- 12747909 TI - Mechanical ventilation and multiple organ failure. PMID- 12747910 TI - Low back pain in rural Tibet. PMID- 12747911 TI - Low back pain in rural Tibet. PMID- 12747912 TI - Stenosis and carotid endarterectomy. PMID- 12747913 TI - Stenosis and carotid endarterectomy. PMID- 12747915 TI - Spanish name indexing errors in international databases. PMID- 12747916 TI - Pesticide regulations in Sri Lanka. PMID- 12747917 TI - Culture and infection control. PMID- 12747918 TI - Culture and infection control. PMID- 12747920 TI - Neonatal hip screening. PMID- 12747919 TI - Childhood cancers and radon. PMID- 12747922 TI - Clever idiot? PMID- 12747921 TI - Neonatal hip screening. PMID- 12747923 TI - Blood transfusion in sickle cell disease: a cautionary tale. PMID- 12747925 TI - Robert Frascino. PMID- 12747928 TI - Heavy metal. PMID- 12747931 TI - Morphological evidence that salivary gland degeneration in the American dog tick, Dermacentor variabilis (Say), involves programmed cell death. AB - During the preoviposition and oviposition periods of ixodid ticks, the salivary glands degenerate. It is unclear whether this is a necrotic or a programmed cell death event. We used an in situ TUNEL technique to determine if salivary gland degeneration involves apoptosis. Salivary glands were dissected from replete females at days 3, 5, 8, 11, 13, and 33 post-detachment. There were no differences in tick weight at detachment, suggesting that changes were not due to engorgement abnormalities. The onset of apoptosis began at day 5 and continued through oviposition at day 33. The greatest amount of nuclei containing fragmented DNA was observed on day 8 post-detachment, suggesting this was the peak occurrence of programmed cell death. Further, the temporal organization of programmed cell death suggests that the granule-secreting acini undergo apoptosis first, and that during the first week of oviposition the type I acini do not exhibit programmed cell death. These data suggest that the type I acini may still function in maintaining off-host hydration state of ovipositing females. These data provide morphological evidence that salivary gland degeneration involves a temporal programmed cell death event. PMID- 12747930 TI - Ultrastructure of the putative stem cell niche in rat mammary epithelium. AB - There is now strong evidence that the stem cells of many tissues reside in specialized structures termed niches. The stem cell niche functions to house and regulate symmetric and asymmetric mitosis of stem cells in mammalian skin, mouse and human bone marrow, mouse brain, gut, and hair follicle, and Drosophila ovary and testis. This regulation is effected through the action of various signaling pathways such as Notch, Hedgehog, Wnt and others. The hormones of the estrous cycle, pregnancy and lactation that initiate growth in mouse mammary epithelium appear to act at a paracrine level to regulate mitosis through Notch receptors. Previous work has established that the putative stem cells of the mammary epithelium in several animal species reside near the basement membrane and never make contact with the ductal lumen. We show that these putative stem cells are found in anatomically specialized places created by the cytoplasmic extensions and modifications of neighboring differentiated cells. Such specializations may help to regulate stem cell activity by modulating molecular traffic to putative stem cells and contact with signaling molecules in the basement membrane. The histological characteristics of these putative niches vary as to the kinds of relationships the cells can have with the basement membrane and neighboring cells and as to how many stem or progenitor cells they may contain. This suggests a plasticity that may be relevant to the response of niches to tissue demands, such as wound healing, the periodic growth and regression of mammary epithelium, the process of mammary tumorigenesis therapeutic strategies for breast cancer. PMID- 12747932 TI - Ovicide-induced serosa degeneration and its impact on embryonic development in Manduca sexta (Insecta: Lepidoptera). AB - Eggs of Manduca sexta treated with the ovicide Ov. 165049 turn orange, and the embryos later die. The orange pigmentation is at first confined to the serosa, and is accompanied by pathological changes of serosal cells. Lipid vesicles aggregate and spindle-shaped electron-lucent vesicles-normally forming a single layer below the apical cell surface-greatly accumulate. The mitochondria swell considerably, and their matrices become electron-lucent. Subsequently, the serosal cells develop additional features of necrosis. They form many autophagic vacuoles which contain mostly degradating mitochondria, but also segregated rough endoplasmic reticulum (rER) and glycogen granules. The whole cytoplasm vesiculates, and the cells shrink considerably. The nuclei become less irregular in shape, the chromatin disperses rather evenly whereas the nucleoli persist. Neither chromatin condensation nor the production of apoptotic bodies was observed-further evidence, that the serosal cells die by necrosis rather than apoptosis. At some stage of development the damaged serosa ruptures, retracts from the embryo and forms a sphere beneath it. It is only after the rupture of the serosa, that the embryo also turns orange and disintegrates rapidly. This shows impressively the protective function which the serosa plays for the embryo. Our physiological tests indicate, that the orange pigmentation of the serosa induced by the ovicide results from a disturbance of the tryptophan/ommochrome pathway serving the excretion of potentially toxic metabolites of tryptophan-rich proteins. The results demonstrate first that the serosa represents an important target for ovicide pesticides and second that it plays a vital role as an excretory organ during embryogenesis. PMID- 12747933 TI - Modes of programmed cell death during Ceratitis capitata oogenesis. AB - In the present study, we demonstrate the existence of two distinct apoptotic patterns in nurse cells during Ceratitis capitata oogenesis. One is developmentally regulated and normally occurs during stages 12 and 13, and the other is stage specific and is sporadically observed during stages 7 and 8. The pre-apoptotic manifestation of the first pattern begins at stage 11 and is characterized by the formation of actin bundles. Subsequently, at stages 12 and 13, the nurse cell nuclei exhibit condensed chromatin and contain fragmented DNA, as revealed by TUNEL assay. The apoptotic nurse cell remnants are phagocytosed by the neighboring follicle cells at the end of oogenesis during stages 13 and 14. In the second apoptotic pattern, which occurs sporadically during stages 7 and 8, the nurse cells degenerate and are phagocytosed by the follicular epithelium that contains apoptotic cell bodies. The data presented herein, compared to previous reported results in Drosophila melanogaster and Dacus oleae (Nezis et al., 2000, 2001), strongly suggest that nurse cell apoptosis is a developmentally regulated and phylogenetically conserved mechanism in higher Dipteran. They also suggest that, the sporadic apoptotic pattern consists of a possible protective mechanism throughout oogenesis when damaged or abnormal egg chambers, are eliminated before they reach maturity. PMID- 12747934 TI - Ultrastructure of the testis in Synbranchus marmoratus (Teleostei, Synbranchidae): the germinal compartment. AB - Synbranchus marmoratus, is a protogynic diandric species in which two types of males, primary and secondary, are found. In both types, the germinal compartment in the testes is of the unrestricted lobular type, but in secondary (sex reversed females) males the lobules develop within the former ovarian lamellae. In the present study, the germinal compartment was examined in both types of males using light microscopy as well as scanning and transmission electron microscopy. Germinal compartment is limited by a basement membrane and contains Sertoli and germ cells. During maturation, processes of Sertoli cells form the borders of spermatocysts containing isogenic germ cells. Characteristically, type A and type B spermatogonia have a single nucleolus and grouped mitochondria associated with dense bodies or nuage. Type B spermatogonia, spermatocytes and spermatids are joined by cytoplasmatic bridges and are confined within spermatocysts. Secondary spermatocytes are difficult to find, indicating that this stage is of short duration. Biflagellated spermatozoa have a rounded head, no acrosome, and possess a midpiece consisting of two basal bodies, each of which produces a flagellum with a typical 9+2 microtubular composition. No associations occur between sperm and Sertoli cells. There were no differences between spermatogenesis in primary and secondary males in this protogynic, diandric fish. PMID- 12747936 TI - Apoptotic cell death of oestrogen activated lactotrophs induced by tamoxifen. AB - Stimulation and inhibition of lactotroph cells cause remarkable morphological and functional changes. In keeping with these changes, the size of the lactotroph cell population undergoes striking alterations due to proliferation or cell death. Factors involved in the induction of apoptosis of pituitary cells are not well established. We demonstrated earlier that oestrogens prevent lactotroph cells of female rats to die by apoptosis induced by bromocryptine treatment, a fact that can be reversed in ovariectomised rats. In this study, we developed experimental models for in vivo and in vitro studies to gain further insight on the survival effect of oestrogens on lactotrophs. In rats pretreated with oestrogens, tamoxifen generates a massive cell death by apoptosis as validated by the TUNEL technique and DNA electrophoresis of pituitary gland. On electron microscope observations, numerous lactotrophs exhibited progressive morphological changes in the nuclei compatible with the apoptotic process. The cells remaining intact also exhibit signs of inhibition due to a significant transformation of regular lactotrophs in atypical subtypes. In pituitary cell cultures exposed to tamoxifen and oestrogen simultaneously, most of the lactotrophs displayed features of apoptosis in the nucleus. The present reports gathered new evidences on the apoptogenic potential of tamoxifen on lactotroph cells, and corroborates the contribution of oestrogens to sustain both a balanced population of lactotrophs and a competent secretory activity. The concept that opposed activities, such as inhibition and stimulation, can activate apoptosis is also strengthen by these observations. PMID- 12747935 TI - Relationship between cardiac protein tyrosine phosphorylation and myofibrillogenesis during axolotl heart development. AB - The axolotl, Ambystoma mexicanum, is a useful system for studying embryogenesis and cardiogenesis. To understand the role of protein tyrosine phosphorylation during heart development in normal and cardiac mutant axolotl embryonic hearts, we have investigated the state of protein tyrosine residues (phosphotyrosine, P Tyr) and the relationship between P-Tyr and the development of organized sarcomeric myofibrils by using confocal microscopy, two-dimensional isoelectric focusing (IEF)/SDS-polyacrylamide gel electrophoresis (PAGE) and immunoblotting analyses. Western blot analyses of normal embryonic hearts indicate that several proteins were significantly tyrosine phosphorylated after the initial heartbeat stage (stage 35). Mutant hearts at stages 40-41 showed less tyrosine phosphorylated staining as compared to the normal group. Two-dimensional gel electrophoresis revealed that most of the proteins from mutant hearts had a lower content of phosphorylated amino acids. Confocal microscopy of stage 35 normal hearts using phosphotyrosine monoclonal antibodies demonstrated that P-Tyr staining gradually increased being localized primarily at cell-cell boundaries and cell-extracellular matrix boundaries. In contrast, mutant embryonic hearts showed a marked decrease in the level of P-Tyr staining, especially at sites of cell-cell and cell-matrix junctions. We also delivered an anti-phosphotyrosine antibody (PY 20) into normal hearts by using a liposome-mediated delivery method, which resulted in a disruption of the existing cardiac myofibrils and reduced heartbeat rates. Our results suggest that protein tyrosine phosphorylation is critical during myofibrillogenesis and embryonic heart development in axolotls. PMID- 12747939 TI - Opioids and the apoptotic pathway in human cancer cells. AB - This study was designed to examine the role of opioids in cell survival, with an emphasis on the mechanism of opioid growth factor (OGF, [Met(5)]-enkephalin) dependent growth inhibition. Using three human cancer cell lines: MIA PaCa-2 pancreatic adenocarcinoma, HT-29 colon adenocarcinoma, and CAL-27 squamous cell carcinoma of the head and neck, and OGF and the opioid antagonist naltrexone (NTX) at a dosage (10(-6)M) selected because it is known to repress or increase, respectively, cell replication, the effects on apoptosis (TUNEL, Annexin V) and necrosis (trypan blue) were investigated on days 2, 5, and 7 of exposure. In addition, the influence of a variety of other natural and synthetic opioids on apoptosis and necrosis was examined at a dosage of 10(-6)M. OGF, NTX, naloxone, [D-Pen(2,5)]-enkephalin, [Leu(5)]-enkephalin, dynorphin A1-8, beta-endorphin, endomorphin-1 and -2, and methadone at concentrations of 10(-6)M did not alter cell viability of any cancer cell line. Exposure of cultures to [D Ala(2),MePhe(4),Glycol(5)]-enkephalin (DAMGO), morphine, or etorphine at 10(-6)M significantly increased the number of adherent cells positively stained for TUNEL and Annexin V, as well as the number of necrotic cells in the supernatant, from control levels at all time points studied. The effects of DAMGO, morphine, and etorphine on apoptosis/necrosis were not fully blocked by concomitant administration of naloxone. Despite the increase in cell death in some opioid treated groups, the number of apoptotic and necrotic adherent cells, and the number of necrotic cells in the supernatant, was no more than 1-2% of the total cell population. These results indicate that the inhibitory (OGF) or stimulatory (NTX) action on cell growth in tissue culture is not due to alterations in apoptotic or necrotic pathways. Moreover, although some opioids increased cell death, and dose-effect relationships need to be established, this activity was not of great magnitude and supports the previously reported lack of growth inhibition of many of these compounds. PMID- 12747940 TI - Control by tachykinin NK(2) receptors of CRF(1) receptor-mediated activation of hippocampal acetylcholine release in the rat and guinea-pig. AB - In vivo microdialysis was employed to explore the effects of different selective non-peptides NK(1),NK(2) and NK(3) receptor antagonists on the corticotropin releasing factor (CRF)-induced release of acetylcholine (ACh) in the hippocampus of rats and guinea-pigs. In both species, the intracerebroventricular (i.c.v.) administration of CRF produced a time- and dose-dependent increase in hippocampal ACh release that was totally suppressed by an intraperitoneally (i.p.) pretreatment with the selective non-peptide CRF(1) receptor antagonist antalarmin (30 mg/kg). Pretreatment with the selective NK(2) receptor antagonist SR48968 (1mg/kg, i.p.) significantly reduced the increase of ACh induced by CRF. In contrast, its low-affinity enantiomer SR48965 (1mg/kg, i.p.) or the NK(1) receptor antagonist, GR205171 (1mg/kg, i.p.) did not exert any antagonist effect. Moreover, administration of the selective NK(3) receptor antagonist SR142801 (1mg/kg, i.p.) did not significantly reduce the CRF-induced hippocampal ACh release in guinea-pigs (the only species studied). The selective activity of SR48968 versus GR205171 or SR142801 indicates that NK(2) receptors play a major role in the control of CRF-induced hippocampal ACh release. Moreover, in freely moving rats, two sessions of stroking of the neck and back of the rat for 30 min, at 90 min intervals, known to be a stressful stimulus, produced a marked and reproducible increase in hippocampal ACh release. This effect was prevented by the administration of the two selective non-peptide CRF1 and NK(2) receptor antagonists antalarmin (30 mg/kg, i.p.) and SR48968 (1mg/kg, i.p.), respectively. This suggests that stress-induced activation of the hippocampal ACh system may be under the control of both endogenously released CRF and NKA, and opens the possibility of the existence of a functional interplay between the pathways containing these peptides as we observed in our experiments on anaesthetized animals. PMID- 12747941 TI - Vasoactive intestinal peptide stimulates proliferation in HT29 human colonic adenocarcinoma cells: concomitant activation of Ras/Rap1-B-Raf-ERK signalling pathway. AB - The vasoactive intestinal peptide (VIP) has been shown to regulate cell proliferation and differentiation in many cell types. We previously reported that this neuropeptide inhibited proliferation in HT29 adenocarcinoma cells cultured in serum-containing medium. In addition, it has been demonstrated that VIP induced a potent stimulation of intracellular cAMP production in these cells cultured either in the absence or in the presence of serum. We also demonstrated that VIP induced phosphorylation of the small GTPase Rap1 in these cancerogenous cells. In the present study, the effects of VIP on the proliferation of HT29 cells cultured in the absence of growth factors and various concomitant signalling events were investigated. Under serum-free conditions VIP stimulates HT29 cell proliferation and induced a time- and concentration-dependent ERK activation. Furthermore, VIP induced the activation of the small GTPase Rap1 and of a 95 kDa isoform of the serine/threonine kinase B-Raf. Ras GTPase is also activated in VIP-stimulated cells. We hypothesize that VIP-induced proliferation in HT29 adenocarcinoma cells may involve a cAMP-Rap1/Ras-B-Raf-ERK signalling pathway. PMID- 12747942 TI - Morphine and cocaine influence on CRF biosynthesis in the rat central nucleus of amygdala. AB - The central nucleus of the amygdala is a CRF-containing limbic brain site which mediates both fear-like and avoidance behaviors; moreover it has been hypothesized that atypical stress responses may contribute to compulsive drug use. Therefore, we studied in rat amygdala the level of CRF mRNA by in situ hybrydization, and the level of the peptide using immunocytochemistry after acute and chronic administration of morphine and cocaine and after their withdrawal. Acute injection of morphine (20 mg/kg i.p.) increased CRF mRNA level, but did not change significantly CRF immunoreactivity in the central nucleus of the amygdala. Chronic morphine administration significantly increased the level of CRF mRNA 3, 24 and 48 h after the last dose. Both, acute and chronic cocaine administration increased CRF mRNA, but the peptide level was decreased only after acute cocaine administration. However, in the late withdrawal (48 h after the last dose of cocaine) both mRNA and the peptide levels tended to decrease. The above data suggest that amygdalar CRF system activity is potently activated after administration of morphine and cocaine, and that activation of this system observed at the time of withdrawal from morphine may be responsible for aversion and anxiety related to these states; therefore a CRF1 receptor may be a target for prospective pharmacotherapies of the withdrawal from abused drugs. PMID- 12747943 TI - Complex array of cytokines released by vasoactive intestinal peptide. AB - A complex mixture of five cytokines has been shown to be released by vasoactive intestinal peptide (VIP). Cytokines were measured in paired samples of culture medium and astroglial cytosol by capillary electrophoresis. This is the first description of VIP-mediated release for TNF-alpha, IL-3, G-CSF and M-CSF from astrocyte cultures. Kinetic studies after VIP treatment demonstrated a gradual but incomplete depletion of cytosolic cytokine levels, with differences observed among the cytokines. Significant increases in release were apparent within 15-30 min for all cytokines. As the recognized VIP receptors (VPAC1 and VPAC2) are linked to adenylate cyclase and also interact with pituitary adenylate cyclase activating polypeptide-38 (PACAP-38), both this homologous peptide and 8-bromo cAMP were investigated and compared to VIP-mediated release. Treatment with 1 mM 8-bromo cAMP produced cytokine release similar in amount to 0.1 nM PACAP-38, but significantly less (<50%) in comparison to 0.1 nM VIP. PACAP-38 and VIP exhibited similar EC(50)'s for the release of G-CSF and TNF-alpha; however, the maximal release was 4-6 times greater for VIP than for PACAP-38. This similarity in potency suggested a VPAC-like receptor; however, the greater efficacy for VIP in comparison to PACAP-38, combined with a lack of cAMP production at subnanomolar concentrations of VIP, suggested a mechanism not currently associated with VPAC receptors. For M-CSF, IL-3 and IL-6, the EC(50)'s of VIP were 3-30 times more potent than those of PACAP-38 in producing release. These studies suggested that multiple mechanisms mediate cytokine release in astrocytes: (1) a low efficacy release produced by PACAP-38 that is cAMP-mediated and (2) a high efficacy, VIP preferring mechanism that was not linked to cAMP. In summary, subnanomolar concentrations of VIP released a complex array of cytokines from astrocytes that may contribute to the mitogenic and neurotrophic properties of this neuropeptide in the central nervous system. PMID- 12747944 TI - The structure of the FMRFamide receptor and activity of the cardioexcitatory neuropeptide are conserved in mosquito. AB - Numerous peptides are structurally related to the cardioexcitatory tetrapeptide FMRFamide. One subgroup of FMRFamide-related peptides (FaRPs) contains an FMRFamide C terminus. Searches of the Drosophila melanogaster genome database identified the first invertebrate FMRFamide G-protein coupled receptor (GPCR), DrmFMRFa-R (Cazzamali and Grimmelikhuijzen, Meeusen et al., 2002). In order to explore molecular mechanisms involved in FMRFamide signal transduction we identified a receptor from the malaria mosquito Anopheles gambiae genome (Holt et al., 2002), AngFMRFa-R, and compared its structure to DrmFMRFa-R. The cytoplasmic loops, extracellular loops, and transmembrane regions are highly conserved between these two FMRFamide receptors. Another subgroup of FaRPs is the sulfakinins which are represented by the consensus structure -XDYGHMRFamide, where X is D or E (Nichols, 2003). We compared AngFMRFa-R and DrmFMRFa-R to the A. gambiae sulfakinin receptors, ASK-R1 and ASK-R2 ( Duttlinger et al., 2003), and the D. melanogaster sulfakinin receptors, DSK-R1 and DSK-R2 Brody and Cravchik, 2000; Hewes and Taghert, 2001 ). The cytoplasmic loops, extracellular loops, and the transmembrane regions are not highly conserved between the FMRFamide and sulfakinin receptors. In order to explore the role of FMRFamide in mosquito biology we measured the effect of the tetrapeptide on in vivo heart rate. The tetrapeptide increased the frequency of spontaneous contractions of the larval mosquito heart and, thus, increased heart rate. These data support the conclusion that the structure of the FMRFamide receptor and activity of the cardioexcitatory FMRFamide neuropeptide are conserved in mosquito. PMID- 12747945 TI - Blockade of dorsal hippocampal kappa-opioid receptors increases blood pressure in normotensive and isolation-induced hypertensive rats. AB - Previous research in our laboratory has established a restraining role of the hippocampal kappa-opioid receptor system in the neural control of blood pressure. Chronic or acute hippocampal administration of kappa-agonists has been shown to reduce blood pressure. Isolation of male Sprague-Dawley rats provokes an increase in blood pressure, which has been proposed as a valid model of mild emotionally induced hypertension. It was the purpose of this study to investigate the blood pressure effects of dorsal hippocampal administration of nor-binaltorphimine (nor BNI), a selective kappa-opioid receptor antagonist; in conscious male Sprague Dawley rats subjected to isolation-induced hypertension. Chronic bi-hippocampal microinjection of nor-BNI (10 nmol per side, twice a day for 13 days) caused increases in systolic blood pressure in grouped rats from a mean of 125 to 148 mmHg, and in isolated rats from a mean of 125 to 151 mmHg. This hypertension was similar in magnitude to the increase observed after rats were isolated for 7 days and treated with vehicle in a similar fashion (mean systolic blood pressure 144 mmHg). The increases in blood pressure were accompanied by bradycardia. No significant responses were seen in the blood pressure of grouped rats treated with vehicle. The hypotensive response to a single hippocampal microinjection of the kappa-agonist U62, 066E (10 or 20 nmol) was prevented in anesthetized rats treated previously with nor-BNI, indicating that in the nor-BNI treated rats there was effective blockade of dorsal hippocampal receptors. These data suggest that the rat hippocampal kappa-opioid receptor system and the endogenous neuropeptide dynorphin may be involved in the central neural regulation of blood pressure. PMID- 12747948 TI - Induction of human NK cell-mediated cytotoxicity by CD40 triggering on antigen presenting cells. AB - Engagement of CD40 on antigen presenting cells (APC) is central to the initiation of cell-mediated immune response. Here, we investigated the ability of CD40 ligation on APC to induce NK cell-mediated cytotoxicity in the human system and the mechanism(s) underlying this process. We showed that APC (consisting in adherent peripheral blood mononuclear cells) (PBMC), pre-stimulated with anti CD40 monoclonal antibodies and co-cultured with autologous non-adherent PBMC for 5-9 days, induced CD3-/CD56+ NK cell-mediated cytotoxicity as well as CD3+/CD56+ T cell-mediated unrestricted cytotoxic activity. The generation of NK cell mediated cytotoxicity was independent on cell-to-cell contact between CD40 triggered APC and NK cells. Moreover, we found that IL-12 did not play a role in NK cells induction by anti-CD40 priming, while IL-2 and IL-15 did play a role. Our results provide an insight into the mechanism by which NK cells are activated in peripheral blood and useful informations for therapeutic application of anti CD40 antibodies. PMID- 12747949 TI - A novel tumour model system for the study of long-term protective immunity and immune T cell memory. AB - We present a novel non-transgenic system to be used for studies on anti-tumour adoptive immunotherapy (ADI) and long-term T cell memory. Tumour-reactive donor immune cells against lacZ-transfected syngeneic tumour cells (ESbL-Gal) were generated from a naive T cell repertoire in DBA/2 mice by a well-established priming/restimulation protocol, and transferred to tumour-inoculated athymic nu/nu mice. The donor immune cells efficiently mediated protective anti-tumour immunity involving both CD4(+) and CD8(+) T cells, and anti-metastatic effects were stronger in 4.5 Gy pre-irradiated than in non-irradiated tumour-inoculated hosts. Long-term persistence of beta-galactosidase (Gal)-specific T cells was shown ex vivo by tetramer staining of CD8(+) T cells specific for an immunodominant Gal epitope. Resistance of treated nu/nu mice against tumour rechallenge revealed the existence of long-term protective immune memory. PMID- 12747950 TI - Immunosuppressive effects of a Kv1.3 inhibitor. AB - The voltage gated potassium channel (Kv1.3) has been shown to play a role in immune responsiveness. Blockade of the channel led to diminution of T cell activation and delayed type hypersensitivity. Previous in vitro studies of the blockade were focused on T cell activation and proliferation. In this study we examined other T and monocytic cell mediated events to glean the extent of the immunosuppressive effects of a Kv1.3 specific inhibitor, Margatoxin (MgTX). We found that MgTX inhibited the intracellular production of Th-1 as well as Th-2 cytokines. MgTX can also inhibit IL-2 production and proliferation of T cells upon stimulation with anti-CD3 and VCAM-1. Furthermore, a redirected cytolytic activity was also inhibited by MgTX. However, MgTX did not inhibit generation of CTL to EBV transformed lymphoma cells or antibody-dependent cellular cytolysis mediated by monocytes. It appears that a Kv1.3 blockade does not affect all immune responses, particularly those of innate immunity. PMID- 12747951 TI - Inhibition of tumor rejection by gammadelta T cells and IL-10. AB - Although many tumors express tumor-specific antigens, most fail to stimulate effective immune responses. Tumors generally lack co-stimulatory molecules, which can lead to tolerance of tumor-specific T cells and progressive tumor growth. Here, we demonstrate that the ovalbumin (OVA) transfected EL4 tumor, E.G7-OVA, grows progressively in syngeneic mice even though the tumor can be rejected if the mice are immunized with OVA in adjuvant. E.G7-OVA grew more rapidly in RAG-1 deficient than sufficient mice suggesting that normal mice make an abortive immune response to this tumor. Depletion of gammadelta T cells or IL-10 augmented the ability of B6 mice to reject E.G7-OVA. Spleen cells from normal, but not IL 10 knockout, mice reconstituted rapid tumor growth in gammadelta T cell-deficient mice. Thus, gammadelta T cells play an important role in preventing immune elimination of this tumor by a mechanism that directly or indirectly involves IL 10. PMID- 12747952 TI - In vitro modulation of cytokine production by lymphocytes in human coccidioidomycosis. AB - The modulation of the cytokine response to coccidioidal antigen by lymphocytes from donors with coccidioidomycosis was examined. In initial experiments, samples from 13 healthy immune donors and seven donors with active coccidioidomycosis anergic to the coccidioidal antigen T27K were assessed for CD3 lymphocyte expression of intracellular IFN-gamma using whole blood analysis. Addition of 10 ng/ml of recombinant IL-12 significantly increased response to T27K among immune and anergic subjects (p<0.05), but the percent of cells expressing IFN-gamma was still significantly greater for immune subjects. Among immune donors, the percentage of CD3 lymphocytes expressing IFN-gamma was significantly reduced with the addition of 10 ng/ml of recombinant IL-4, IL-10, TGF-beta, or their combination (for all, p<0.05). Among anergic donors, addition of 10 ng/ml of anti IL-10 significantly increased IFN-gamma production (p<0.05), but addition of anti IL-4 or anti-TGF-beta did not. Among immune donors, the percent of both CD3 lymphocytes and NK cells expressing IFN-gamma after 24h of T27K was increased above control (p<0.05), while the percent of NK cells producing TNF-alpha in response to T27K was not greater than control. Depletion of NK cells from peripheral blood mononuclear cells resulted in significant increases in TNF-alpha and IL-10 (for both, p<0.05) but resulted in no significant decrease in IFN-gamma or IL-2. These data demonstrate a differential response to stimulation with the coccidioidal antigen T27K among donors with coccidioidomycosis that can be manipulated by cell type and cytokine environment. PMID- 12747953 TI - Phosphorylation of activating transcription factor in murine splenocytes through delta opioid receptors. AB - Delta opioid receptors (DORs) modulate TCR signaling through the mitogen activated protein kinases (MAPKs), ERKs 1 and 2. These studies determined whether a DOR agonist alone ([D-Ala(2)-D-Leu(5)]enkephalin; DADLE) affects phosphorylation of the activating transcription factor (ATF-2) and its interaction with the MAPK, c-Jun NH(2)-terminal kinase (JNK). DOR expression was induced on murine splenocytes by anti-CD3 and then quiescent cells were treated with DADLE. DADLE, itself, dose-dependently induced maximal phosphorylation of ATF-2 within 5-10min; naltrindole, a specific antagonist, abolished this. Anti ATF-2 immunoprecipitates from control and DADLE-treated splenocytes showed a dominant 59kDa phosphorylated band and a 71kDa band. DADLE stimulated phosphorylation of both bands, although the 71kDa band was selectively immunoprecipitated by anti-JNK. Thus, DADLE stimulated phosphorylation of 71kDa ATF-2 and its association with JNK, suggesting that JNK is activated through DORs. Along with previous observations, these studies suggest that lymphocyte DORs can affect the activation of MAPKs by TCR-independent stimulation (e.g., JNK) or indirectly by modulating TCR-dependent stimulation (e.g., ERK). PMID- 12747954 TI - Quantitative interplay between activating and pro-apoptotic signals dictates T cell responses. AB - Antigen-presenting cells (APC) can express surface ligands with both T cell activating and inhibitory capacities, prompting the question of how responding T cells integrate opposing trans signals concurrently delivered by APC. To address this question in a quantitative fashion, we turned to protein transfer as a unique experimental approach that is well-suited for addressing such questions from a quantitative standpoint. Costimulatory (either B7-1*Fc(gamma1) or Fc(gamma1)*4-1BBL) and pro-apoptotic (Fc(gamma1)*FasL) Fc fusion proteins were quantitatively "painted" in varying ratios onto surrogate APC pre-coated with palmitated-protein A, the latter serving as a surface anchor. Evaluating the signaling potential of these various painted cells in a standard in vitro T cell proliferation assay, we demonstrated that at a given level of TCR triggering, the quantitative balance between costimulator (B7-1 or 4-1BBL) and FasL dictates the magnitude of the proliferative T cell response. Furthermore, when the costimulator density is kept constant, there is also a quantitative balance between TCR-directed and FasL signals. Interesting species-specific nai;ve versus memory T cell subset differences emerged with regard to susceptibility to Fas mediated apoptosis and costimulator:FasL opposition. Taken together, these data demonstrate for the first time a quantitative interplay between activating and pro-apoptotic trans signals that dictates the magnitude of T cell responses. PMID- 12747955 TI - Does complexity belie a simple decision--on the Efroni and Cohen critique of the minimal model for a self-nonself discrimination. AB - The immune system somatically generates a large and random paratopic repertoire that must be sorted into those specificities (anti-self) which, if expressed, would debilitate the host and those specificities (anti-nonself) which, if not expressed, would leave the host unprotected from infection. The critique of Efroni and Cohen that minimal models are misleading and without heuristic value is evaluated by illustrative examples. PMID- 12747956 TI - Morphological and immunological evidence of a unique selective production and endoplasmic reticular accumulation of interleukin-1alpha in rat peritoneal macrophages induced by Pseudomonas aeruginosa exotoxin A. AB - The immunotoxicity of Pseudomonas aeruginosa exotoxin A (ETA) on macrophages was evaluated by incubating rat peritoneal macrophages (RPM) with 1-100 ng/ml ETA for 3-60 h. Although the overall changes in cell viability and DNA, RNA, and protein synthesis of the ETA-treated RPM (E-RPM) were reduced in a dose- and time dependent manner, there was a transient but evident rebound in RNA and/or protein synthesis at 24-36 h post-incubation (HPI) at 1-50 ng/ml ETA. However, a more apparent enhancement appeared in RNA and protein synthesis at 36-48 HPI in 10 and 50 ng/ml E-RPM after normalized on the basis of viable cell. Most 50-100 ng/ml E RPM underwent necrosis/apoptosis before 24 HPI. By 36 HPI, 41% of 10 ng/ml E-RPM remained viable but were full of cytoplasmic granules due to the accumulation of glycoprotein in segmentally dilated endoplasmic reticulum. Immunological staining of the granules revealed strong IL-1alpha but weak or no signals for IL-1beta, IL 1 receptor antagonist, IL-6, and TNF-alpha. A time-dependent increase in IL 1alpha but no IL-1beta was detected in cell lysate of 10 ng/ml E-RPM; however, neither IL-1alpha nor IL-1beta was detected in culture supernatant. Thus, besides cytopathic and functional effects, ETA could induce a unique selective production and endoplasmic reticular accumulation of IL-1alpha in RPM. PMID- 12747957 TI - Endothelial hyperpermeability in vascular leakage. PMID- 12747958 TI - Role of Ca2+ signaling in the regulation of endothelial permeability. AB - The vascular endothelial cell forms a semipermeable barrier between blood and interstitium. Inflammatory mediators such as thrombin and histamine induce vascular leakage defined as increased endothelial permeability to plasma proteins and other solutes. Increased endothelial permeability is the hallmark of inflammatory vascular edema. Inflammatory mediators that bind to heptahelical G protein-coupled receptors (GPCR) trigger increased endothelial permeability by increasing the intracellular Ca(2+) concentration ([Ca(2+)](i)). The rise in [Ca(2+)](i) activates key signaling pathways, which mediate cytoskeletal reorganization (through myosin light chain (MLC)-dependent contraction) and disassembly of VE-cadherin at the adherens junctions. The Ca(2+)-dependent protein kinase C (PKC) isoform, PKC-alpha, plays a critical role in initiating endothelial cell contraction and disassembly of VE-cadherin junctions. The increase in [Ca(2+)](i) induced by a variety of agonists is achieved by the generation of inositol 1,4,5-trisphosphate (IP3), activation of IP3 receptors (IP3R), release of stored intracellular Ca(2+), and Ca(2+) entry through plasma membrane channels. Recent findings demonstrate that IP3-sensitive Ca(2+) store depletion activates plasma membrane cation channels (i.e., store-operated cation channels (SOC) or Ca(2+) release activated channels) to cause Ca(2+) influx in endothelial cells. This mode of Ca(2+) influx is also known as capacitative Ca(2+) entry (CCE). Store-operated Ca(2+) influx signals increase in permeability and nitric oxide (NO) production and provokes changes in gene expression in endothelial cells. Recent studies have established that the Drosophila transient receptor potential (TRP) gene family of channels expressed in endothelial cells can function as SOC. Deletion of one of the TRP homologues, TRPC4, in mouse caused impairment in store-operated Ca(2+) current and Ca(2+) store release activated Ca(2+) influx in aortic and lung endothelial cells (LEC). In TRPC4 knockout (TRPC4(-/-)) mice, acetylcholine-induced endothelium-dependent smooth muscle relaxation was drastically reduced. In addition, TRPC4(-/-) mice LEC exhibited lack of actin stress fiber formation and cell retraction in response to thrombin activation of proteinase-activated receptor-1 (PAR-1) in endothelial cells. The increase in lung microvascular permeability in response to thrombin receptor activation was inhibited in TRPC4(-/-) mice. These results indicate that endothelial TRP channels such as TRPC1 and TRPC4 play an important role in signaling the increase in endothelial permeability. PMID- 12747959 TI - Rho GTPases and the regulation of endothelial permeability. AB - Endothelial permeability depends on the integrity of intercellular junctions as well as actomyosin-based cell contractility. Rho GTPases have been implicated in signalling by many vasoactive substances including thrombin, tumour necrosis factor alpha (TNF-alpha), bradykinin, histamine, lysophosphatidic acid (LPA), vascular endothelial growth factor (VEGF), and hepatocyte growth factor (HGF). Two Rho family GTPases, Rho and Rac, have emerged as key regulators acting antagonistically to regulate endothelial barrier function: Rho increases actomyosin contractility, which facilitates breakdown of intercellular junctions, whereas Rac stabilizes endothelial junctions and counteracts the effects of Rho. In this review, we present evidence for the opposing effects of these two regulatory proteins and discuss links between them and other key signalling molecules such as cyclic AMP (cAMP), cyclic GMP (cGMP), phosphatidylinositide 3 kinases (PI3Ks), mitogen-activated protein kinases (MAPKs), and protein kinases C (PKCs). We also discuss strategies for targeting Rho GTPase signalling in therapies for diseases involving altered endothelial permeability. PMID- 12747960 TI - Role of tyrosine kinase signaling in endothelial cell barrier regulation. AB - Phosphorylation of proteins on tyrosine acts as a reversible and specific trigger mechanism, forming or disrupting regulatory connections between proteins. Tyrosine kinases and phosphatases participate in multiple cellular processes, and considerable evidence now supports a role for tyrosine phosphorylation in vascular permeability. A semipermeable barrier between the vascular compartment and the interstitium is maintained by the integrity of endothelial monolayer, controlling movement of fluids, macromolecules and leucocytes. Barrier function is regulated by the adjustment of paracellular gaps between endothelial cells (ECs) by two antagonistic forces, centripetal cytoskeletal tension and opposing cell-cell and cell-matrix adhesion forces. Both cytoskeletal filaments and adhesion sites are intimately linked in complex machinery which is regulated by multiple signaling events including protein phosphorylation and/or protein translocation to specific intracellular positions. Tyrosine kinases occupy key positions in the mechanism controlling cell responses mediated through various cell surface receptors, which use tyrosine phosphorylation to transduce extracellular signal. PMID- 12747961 TI - Protein kinase signaling in the modulation of microvascular permeability. AB - The permeability of exchange microvessels is regulated through complex interactions between signaling molecules and structural proteins in the endothelium. Endothelial barrier integrity is maintained by adhesive interactions occurring at the cell-cell and cell-matrix contacts via junctional proteins and focal adhesion complexes that are anchored to the cytoskeleton. Cyclic AMP (cAMP) and cAMP-dependent kinase counteract with the nitric oxide (NO)-cyclic GMP (cGMP) pathway to protect the basal barrier function. Upon stimulation by physical stress, growth factors, or inflammatory agents, endothelial cells undergo a series of intracellular signaling reactions involving activation of protein kinase C (PKC), protein kinase G (PKG), mitogen-activated protein kinases (MAPK), and/or protein tyrosine kinases. The phosphorylation cascades trigger biochemical and conformational changes in the barrier structure and ultimately lead to an opening of the paracellular pathway. In particular, myosin light chain kinase (MLCK) activation and subsequent myosin light chain (MLC) phosphorylation in endothelial cells directly result in cell contraction and shape changes. The phosphorylation of beta-catenin may cause disorganization of adherens junctions or dissociation of vascular endothelial (VE)-cadherin-catenin complex from its cytoskeletal anchor, leading to loose or opened intercellular junctions. Additionally, focal adhesion kinase (FAK) phosphorylation-coupled focal adhesion assembly and redistribution provide an anchorage support for the conformational changes occurring in the cells and at the cell junctions. The Src family tyrosine kinases may serve as common signals that coordinate these molecular events to facilitate the paracellular transport of macromolecules. The critical roles of protein kinases in endothelial hyperpermeability implicate the therapeutic significance of protein kinase inhibitors in the prevention and treatment of diseases and injuries that are associated with microvascular barrier dysfunction. PMID- 12747962 TI - Regulation of vascular permeability by vascular endothelial growth factors. AB - Increased vascular permeability is one of the first stages in both physiological and pathological angiogenesis-the generation of new blood vessels from preexisting vasculature. Although this has been hypothesised to be true in physiological angiogenesis, it is clearly a mark of blood vessel growth in disease. Normal, healthy blood vessel growth (physiological angiogenesis) occurs throughout development as well as during tissue repair and growth in adult tissues. Angiogenesis is also seen in a wide variety of diseases, which include all the major causes of mortality in the West-heart disease, cancer, stroke, vascular disease, and diabetes. Much of this angiogenesis is significantly different from normal blood vessel growth and is termed pathological angiogenesis. Angiogenesis is regulated by vascular growth factors, the most notable being the vascular endothelial growth factor family of proteins (VEGF). These act on specific receptors in the vascular system to stimulate new vessel growth by a number of mechanisms. VEGFs also directly stimulate increased vascular permeability to water and large molecular weight proteins and vasodilatation. These two effects result in a large flux of water and macromolecules from the vasculature to the interstitium, often resulting in oedema. This review will outline the mechanisms by which VEGFs do this and discuss some of the difficulties in interpreting data from VEGF studies due to the conflicting and synergistic effects of these actions. PMID- 12747963 TI - Role of interendothelial adhesion molecules in the control of vascular functions. AB - The function of endothelium is the lining of the vessel wall and the control of vascular permeability, homeostasis and leukocyte emigration from the blood into the surrounding tissue. Different adhesion molecules expressed in a coordinated and regulated way control this function. In this review, we discuss adhesion molecules involved in endothelial junctions and their involvement in leukocyte transendothelial migration. Passage of the leukocyte across the endothelium appears to require delocalization of certain vascular adhesion molecules whereas other molecules interact directly with leukocyte ligands. Understanding of the function of vascular adhesion molecules is further complicated as they transduce signals to the endothelium and interact with the cytoskeleton and adaptor proteins. PMID- 12747964 TI - Vascular pharmacology of acute lung injury and acute respiratory distress syndrome. AB - Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) following sepsis, major trauma and surgery are leading causes of respiratory insufficiency, warranting artificial ventilation in the intensive care unit. It is caused by an inflammatory reaction in the lung upon exogenous or endogenous etiologies eliciting proinflammatory factors, and results in increased alveolocapillary permeability and protein-rich alveolar edema. The interstitial and alveolar inflammation and edema alter ventilation perfusion matching, gas exchange and mechanical properties of the lung. The current therapy of the condition is supportive, paying careful attention to fluid balance, relieving the increased work of breathing and improving gas exchange by mechanical ventilation, but in vitro, animal and some clinical research is done to evaluate the value of anti inflammatory therapies on morbidity and outcome, including inflammatory cell stabilizing corticosteroids, xanthine derivates, prostanoids and inhibitors, O(2) radical scavenging factors such as N-acetylcysteine, surfactant replacement, vasodilators including inhaled nitric oxide, vasoconstrictors such as almitrine, and others. None of these compounds has been proven to benefit survival in patients, however, even though carrying a physiologic benefit, except perhaps for steroids that may improve outcome in the later stage of ARDS. This partly relates to the difficulty to assess the lung injury at the bedside, to the multifactorial pathogenesis and the severity of comorbidity, adversely affecting survival. PMID- 12747965 TI - Targets for pharmacological intervention of endothelial hyperpermeability and barrier function. AB - Many diseases share the common feature of vascular leakage, and endothelial barrier dysfunction is often the underlying cause. The subsequent stages of endothelial barrier dysfunction contribute to endothelial hyperpermeability. Vasoactive agents induce loss of junctional integrity, a process that involves actin-myosin interaction. Subsequently, the interaction of leukocytes amplifies leakage by the leukocyte-derived mediators. The processes mainly occur at the postcapillary venules. The whole microvascular bed, including the capillaries, becomes involved in vascular leakage by the induction of angiogenesis. Plasma leakage results from gaps between endothelial cells as well as by the induction of transcellular transport pathways. Several mechanisms can improve endothelial barrier function, depending on the tissue affected and the cause of hyperpermeability. They include blockade of specific receptors and elevation of cyclic AMP (cAMP) by agents such as beta(2)-adrenergic agents. However, current therapies based on these principles often fail. Recent research has identified several new promising targets for pharmacological therapy. Endogenous compounds were also found with barrier-improving characteristics. Important insights were obtained in the different pathways involved in barrier dysfunction. Such insights regard the regulation of endothelial contraction and endothelial junction integrity: inhibitors of RhoA activation and Rho kinase represent a potentially valuable group of agents with endothelial hyperpermeability reducing properties, and strategies to target vascular endothelial growth factor (VEGF)-mediated edema are under current investigation. In clinical practice, not only tools to improve an impaired endothelial barrier function are necessary. Sometimes, a controlled, temporal, and local increase in permeability can also be desired, for example, with the aim to enhance drug delivery. Therefore, vessel leakiness is also being exploited to enable tissue access of liposomes, viral vectors, and other therapeutic agents that do not readily cross healthy endothelium. This review discusses strategies for targeting signaling molecules in therapies for diseases involving altered endothelial permeability. PMID- 12747966 TI - Changing concepts in the surgical management of the cervical node metastasis. PMID- 12747967 TI - A non-invasive technique for studying oral epithelial Epstein-Barr virus infection and disease. AB - Oral Epstein-Barr virus (EBV) infection is associated with hairy leukoplakia and possibly other oral diseases. Many studies of oral EBV infection utilize surgical specimens. This study tested a non-invasive brush biopsy technique as an alternative to surgical biopsy to study oral EBV infection and disease. Paired, same-site, samples of tongue epithelium were obtained from research subjects, first by brush and then by surgical biopsy. Brush cells and surgical specimens were fixed and prepared for histologic sectioning and/or processed for nucleic acid extraction. Brush cell pellet sections proved equivalent to surgical specimen tissue sections for hairy leukoplakia diagnosis by routine histologic staining and EBV immunohistochemistry or in situ hybridization. Amplification of EBV sequences demonstrated superiority of the brush cells over surgical specimens for both sensitivity (90% vs. 73%) and negative predictive value (93% vs. 82%). This non-invasive brush biopsy technique should facilitate larger, prospective studies of oral EBV infection and pathogenesis. PMID- 12747968 TI - Gene expression of telomerase related proteins in human normal oral and ectocervical epithelial cells. AB - We analyzed telomerase activities and gene expressions of telomerase components: hTERT, hTR, hTEP1, telomeric repeat binding factors: TRF1, TRF2, and c-myc, Max and Mad in human normal oral and ectocervical epithelial keratinocytes, comparing with those of squamous carcinoma cells and HPV16- or SV40-immortalized cells. Significant telomerase activity and hTERT expression were detected in primary keratinocytes. However, both were dramatically down-regulated during serial passages. The down-regulation of hTERT mRNA was associated with augmented expression of TRF1. Expression of c-myc was slightly decreased, whereas Mad was expressed in parallel with that of hTERT during passages. We also detected an alternate splicing of hTERT transcript in two of four cancer cells and normal aged epithelial cells. These results suggest that the senescence of normal oral and ectocervical keratinocytes is accompanied with up-regulation of TRF1 and down regulation of telomerase activity due to transcriptional suppression of active form of hTERT in vitro. PMID- 12747969 TI - Analysis of epithelial-myoepithelial carcinoma based on the establishment of a novel cell line. AB - Epithelial-myoepithelial carcinoma of the salivary gland is a rare, low-grade, neoplasm, composed of ductal and myoepithelial cells. We present two novel cell lines, which have been characterised by immunofluorescence, derived from an epithelial-myoepithelial carcinoma of the parotid gland. A resected mass of the parotid gland was diagnosed as an epithelial-myoepithelial carcinoma by routine histological examination. Part of the specimen was labelled with a panel of antibodies confirming the tumour type. The other part was finely minced and the explants were incubated in DMEM supplemented with penicillin and streptomycin, at 37 degrees C in a humidified 5% CO(2) atmosphere. Two cell types were identified by immunofluorescence-a small cobblestone cell, positive for AE1/AE3 and p53, and a polyhedral cell, positive for vimentin, smooth muscle markers and S-100. Herein two cell lines are presented in order to open up possibilities of new studies and a discussion of the events that culminate in this bimodal neoplasm is also performed. PMID- 12747971 TI - Is there a natural limit of the transformation rate of oral leukoplakia? AB - Recent reviews described a cumulative risk of 0-38% of oral leukoplakias (OL) to transform into oral squamous cell carcinoma (OSCC). The annual transformation rate (ATR) of OL into OSCC amounted up to 6.3%. To elucidate epidemiological consequences of varying ATR. Variations of the prevalence (1-2%) and ATR (0.5-4%) of OL were used to compute the proportional incidences of OSCC developing in OL. These were compared to the actual incidences of OSCC excerpted from European cancer registries. A ratio <50% of predicted to actual incidence overall indicated a plausible ATR. With an ATR>or=3%, the predicted incidences of OSCC would exceed the reported incidences in all countries. With an ATR47 days: 55%, P=0.03), Ki-67 LI (20%: 56%, P=0.006). A significant prognostic impact on locoregional control was noted for the duration of RT (P=0.01), tumor site (P=0.02), and the Ki-67 LI (P=0.02). A low apoptotic index together with higher proliferation rates led to unfavourable local control as low as 25% compared to the patients with higher apoptotic index (70-80%, P=0.009). An imbalance between apoptotic index and proliferation may identify patients with squamous cell carcinoma at high risk for local recurrence after surgery and postoperative RT. Prospective observation of these factors in clinical trials is warranted to further elucidate this phenomenon. PMID- 12747972 TI - Expression of cyclin A is related to progression of oral squamous cell carcinoma in Taiwan. AB - Cyclin A is required for DNA synthesis during the S phase and progression through the G2/M transition. Increased expression of cyclin A protein has been correlated with poor prognosis in a variety of human tumors. To investigate the possible influence(s) of cyclin A protein on the progression and prognosis of oral squamous cell carcinomas (SCCs) in Taiwan. We examined the expression of cyclin A in oral SCC, epithelial dysplasia (ED) and normal oral mucosa (NOM) by immunohistochemistry using antibodies to cyclin A. Results and Conclusions. The mean labeling indices (LI) in NOM, ED and SCCs were 7.0+/-3.1%, 12.1+/-3.9% and 21.3+/-12.3%, respectively. The cyclin A LI for oral SCCs was significantly higher than that for NOM (P=0.002) or ED (P<0.001). In addition, a high LI for cyclin A was found to correlate with advanced stage (P=0.0048), larger tumor size (P=0.0017), lymph node involvement (P=0.0006) and cancer recurrence (P<0.0001). The Kaplan-Meier analysis showed patients with tumors containing more than 15% cyclin A-positive cells had significantly shorter overall survival than those with tumors containing less than 15% cyclin A-positive cells (P<0.00001). These results indicate that overexpression of cyclin A protein is associated with tumor progression and patient prognosis for oral SCC. PMID- 12747973 TI - Epoxide hydrolase genotype and orolaryngeal cancer risk: interaction with GSTM1 genotype. AB - The human microsomal epoxide hydrolase (EH) gene contains polymorphic alleles which are associated with altered EH activity and may be linked to increased risk for tobacco-related cancers. The objective was to examine the role of EH polymorphisms in orolaryngeal cancer risk. The prevalence of the EH codons 113 and 139 polymorphisms were examined in 81 African American and 142 Caucasian incident orolaryngeal cancer patients and 335 controls frequency-matched on age, sex, and race. In Caucasians, a significant risk increase was observed for subjects with the EH(113Tyr) variant (OR=2.1, 95% CI=1.1-4.0) and predicted high activity EH genotypes in heavy-smokers (>or=35 pack-years; OR=3.4, 95% CI=1.2 9.6). A significant association between predicted high EH activity genotypes and orolaryngeal cancer risk was observed in Caucasian subjects with the GSTM1 null (OR=3.5, 95% CI=1.3-9.3) but not GSTM [+] (OR=0.9, 95%CI=0.4-2.1) genotype. These results suggest that EH polymorphisms play an important role in risk for orolaryngeal cancer in Caucasians. PMID- 12747974 TI - Histomorphologic and morphometric changes in minor salivary glands of the rat tongue during 4-nitroquinoline-1-oxide-induced carcinogenesis. AB - 4-Nitroqinoline-1-Oxide (4NQO)-induced tongue carcinogenesis in rats is considered to be a preferred model for study of oral squamous cell carcinoma. Aim of study was to investigate histomorphologic and morphometric 4NQO-induced changes in tongue minor salivary glands. Histopathological examinations of serous and mucous acini and ducts of tongue salivary glands of 26 Wistar-derived rats were performed after 14 (T(1)), 22 (T(2)) and 28 (T(3)) weeks of 0.001% 4NQO administration in drinking water and compared with nine controls. Histomorphological findings were recorded as normal/abnormal acini and as normal/dysplastic ducts. Morphometrical results were expressed as volume fraction (Vv%) of each of the components. Morphometric and histomorphologic changes in the salivary glands were evident only at T(3) and they included a significant (P=0.008) decrease in the Vv of the serous acini compared with the control group accompanied by abnormal acini (Vv=18%). In contrast, mucous acini and ducts did not demonstrate significant changes. In one case (3.8%), dysplastic ducts were found adjacent to islands of invasive squamous cell carcinoma of tongue mucosa origin. The change in saliva composition expected after considerable damage of the serous glands could create a microenvironment that makes entrapment of the carcinogen easier and prolongs exposure of tongue epithelium. Furthermore, the dysplastic changes in the ducts can serve as a reservoir of carcinoma cells. These observations should be considered in human patients diagnosed with oral dysplasia or carcinoma, especially involving the tongue and floor of mouth. PMID- 12747975 TI - Cyclooxygenase-2 (COX-2) expression in high-risk premalignant oral lesions. AB - Emerging data indicate a link between genetic instability and up-regulation of cyclooxygenase-2 (COX-2). To see if individuals at high risk of oral cancer are candidates for treatment with selective COX-2 inhibitors (coxibs), levels of COX 2 expression in healthy, premalignant and cancerous oral mucosa were compared with the occurrence of DNA ploidy status as a genetic risk marker of oral cancer. COX-2 gene product was evaluated immunohistochemically in 30 healthy persons, in 22 patients with dysplastic lesions without previous or concomitant carcinomas, and in 29 patients with oral carcinomas. The immunohistochemical findings were verified by western blotting. COX-2 expression was correlated to DNA content as a genetic risk marker of oral cancer. COX-2 was up-regulated from healthy to premalignant to cancerous oral mucosa. Thus, COX-2 expression was found in 1 case of healthy oral mucosa (3%). All specimens from healthy mucosa had a normal DNA content. In patients with premalignancies. In 29 patients with oral carcinomas, cyclooxygenase-2 expression was observed in 26 (88%), and aneuploidy was observed in 25 cases (94%, P=0.04). Notably, of 22 patients with dysplastic lesions, COX-2 was exclusively expressed in a subgroup of nine patients (41%) identified to be at high risk of cancer by the aberrant DNA content of their lesions. Seven of these patients were followed for 5 years or more. An oral carcinoma developed in six of them (85%; P=0.02). These findings emphasize the need to determine whether coxibs can reduce the risk of oral cancer in patients with high-risk precancerous lesions. PMID- 12747976 TI - Correlation of P-cadherin and beta-catenin expression and phosphorylation with carcinogenesis in rat tongue cancer induced with 4-nitroquinoline 1-oxide. AB - Using biochemical and immunohistochemical techniques, we have investigated P cadherin, beta-catenin, c-src and c-met protein expression, and phosphorylation of beta-catenin in a rat model of tongue cancer induced with 4-nitroquinoline 1 oxide. Six-week-old male Sprague-Dawley rats were given either normal drinking water (controls) or 50 ppm 4NQO solution as drinking water for 16 and 20 weeks. This treatment produced dysplasia and well-differentiated squamous cell cancer in rat tongues after 16 and 20 weeks, respectively. In controls, P-cadherin and beta catenin were expressed only in cell membranes of tongue suprabasal epithelial cells, whereas strong reaction to P-cadherin antibody was observed during carcinogenesis, especially in nests of cancer cells. However, dysplastic and cancer cells expressed beta-catenin not only in cell membranes but also in the nuclear and cytoplasmic compartments. During carcinogenesis, immunohistochemical reaction to phosphotyrosine increased gradually. Reaction to the c-src product was strongest at the dysplastic stage and, to the c-met product, at the cancer stage. In addition, western blotting analysis showed a marked increase in the expression of beta-catenin and phosphotyrosine in dysplastic and cancer cells compared with the controls. Using immunoprecipitation and western blotting techniques, we found that phosphorylated beta-catenin gradually increased during carcinogenesis. These experiments demonstrate that cell-cell adhesion in epithelial cells was reduced by phosphorylation of beta-catenin and that beta catenin overexpression in nuclear and cytoplasmic compartments during carcinogenesis and the production of the c-met product that is associated with the phosphorylation of beta-catenin in tongue cancer. PMID- 12747977 TI - Establishment of an oral squamous cell carcinoma cell line with high invasive and p27 degradation activities from a lymph node metastasis. AB - The extent of lymph node metastasis is a major determinant in the prognosis of oral squamous cell carcinoma (OSCC). We present here a new OSCC cell line, MSCC 1, established from a lymph node metastasis of a patient with OSCC of gingiva. First, we examined the expression of p27, p53 and Ki-67 in non-neoplastic mucosa, primary and metastatic cancer lesions by immunohistochemistry. Metastatic cancer cells in the lymph node showed the reduced expression of p27 in comparison with cancer cells in the primary lesion. Cancer celLs both in the primary and metastatic lesions showed overexpression of p53 and Ki-67. Overexpression of p53 and reduced expression of p27 in MSCC-1 cells were also determined by western blot analysis. To characterize MSCC-1 cells, furthermore, we examined the invasive activity and cell proliferation of MSCC-1, comparing with those of other OSCC cell lines, HSC-2 and HSC-3 cells. The invasive capacity of MSCC-1 cells was significant higher than HSC-2 and HSC-3 cells, but cell growth of MSCC-1 cells was slower than HSC-2 and HSC-3 cells. Moreover, we examined the p27 degradation activity by in vitro degradation assay. Interestingly, MSCC-1 cells have the strongest p27 degradation activity among the OSCC cell lines examined. In the present study, we newly established MSCC-1 cells with strong invasiveness and p27 degradation activity from a metastatic lesion. These findings suggest that high activity of p27 degradation may concern with invasiveness of OSCC cells and that MSCC-1 cells can be a useful cell model for studying the detailed mechanism of p27 degradation, invasion and metastasis of OSCC. PMID- 12747978 TI - Clinicopathological and immunohistochemical analysis of twenty-five head and neck osteosarcomas. AB - The aim of this study was to analyze the clinicopathological features and immunohistochemical expression of p53, MDM2, CDK4, PCNA and Ki67 proteins in 25 head and neck osteosarcomas registered in a single institution. The mean age of the patients was 29 years and the most common site was the mandible (60%). The predominant histological type was the chondroblastic (72%) and 52% of the cases were classified as intermediate-grade of malignancy. The immunohistochemical analysis displayed positivity in 52% of the cases for p53, 24% for MDM2, 84% for CDK4, 92% for PCNA and 88% for Ki-67. The majority of cases were treated with surgery alone or associated with chemotherapy. Five patients developed local recurrences, four had distant metastasis and six had persistent disease after initial treatment. The overall 5-year and 10-year survival rates were 59% and 49%, respectively, and the most important prognostic factors were prior history of radiation exposure and osteoblastic histological type. PMID- 12747979 TI - The use of sentinel node biopsy in the management of epitheloid haemangioendothelioma of the lip. AB - This report describes a rare tumour, an epitheloid haemangioendothelioma affecting the lower lip. This tumour has a predilection for the head and neck region in young adults. Its potential to metastasise is well recognised, but the likelihood of this is currently uncertain. Current management is usually to locally excise the tumour and follow up, although there is a recognised risk that subsequent presentation with metastatic nodal disease can occur. We present a case occurring in the lip of an 18-year-old girl who had a sentinel node biopsy performed as a staging tool in conjunction with excision of a local recurrence. Although clinical examination and CT imaging of the head and neck found no evidence of metastatic disease, the sentinel node was found to contain metastatic tumour. The result of this unexpected finding was that she was investigated further with additional CT scanning of her chest and abdomen. Subsequently, a therapeutic modified radical neck dissection preserving the accessory cranial nerve was undertaken. After 3 years she remains well with no evidence of recurrent tumour. We believe that the consequence of undergoing sentinel node biopsy, which detected early metastatic tumour and her subsequent treatment, suggests a role for sentinel node biopsy in the management of epitheloid haemangioendothelioma. PMID- 12747981 TI - [Do we need office for anaesthesiologist?]. PMID- 12747980 TI - Dermoscopy in the management of pigmented lesions of the oral mucosa. PMID- 12747982 TI - [Survey of office space and computer availability offered to full time practitioners in departments of anaesthesia and intensive care of French university hospitals]. AB - INTRODUCTION: Office space and computer facilities offered to medical practitioners in departments of anaesthesia (DA) belonging to university hospitals in metropolitan France in 2002 were surveyed. METHOD: A questionnaire was mailed to the 72 heads of DA belonging to university hospitals in metropolitan France in order to assess: (1) the number of full time anaesthesiologists sharing each office; (2) whether a computer was provided to all full time anaesthetists who required one; (3) the adequacy of the offices in terms of the DA's needs; (4) the subjective appreciation of the comfort level of the DA office space when compared to other departments within the institution; (5) whether an office space with a computer was specifically reserved for fellows. RESULTS: Sixty-two replies were received (86.1%). Among full time anaesthesiologists surveyed:only 21.8% occupied an office alone; 1.2% had no office; 36.5, 21.7, 8.2, 3.4, 3.9, 3.2% shared one office with 1, 2, 3, 4, 5, more than 5 colleagues, respectively; 25.8% had a personal computer. Fifty percent of DA surveyed did not reserve a specific office for fellow's need; 75.8% of the offices surveyed were evaluated as being of inadequate comfort level; 64.5% of the offices surveyed were evaluated at a lower comfort level when compared to the office space of other departments within the institution. CONCLUSION: A high response rate was obtained. DA were found to be insufficiently provided with offices and computer facilities in french university hospitals. Such a situation, which is both surprising and questionable in an industrialised country, warrants a debate. PMID- 12747983 TI - [Evaluation of 5 years of postoperative pain management in orthopaedic surgery, in a private hospital, following quality standard management]. AB - OBJECTIVE: To report the 5 years' experience of a postoperative pain service according to quality standards management. MATERIAL AND METHODS: The goals of pain treatment were determined from the description of the initial inquiries performed in 1994. Decrease in the postoperative pain for all the patients operated at the first and the third postoperative day, systematic evaluation of the pain, implementation of common therapeutic protocols, information and satisfaction of the patients, improvement in side effects' management, and a bearable managing cost. The deadline of these objectives was 1999. Patients were followed-up by a questionnaire at the third postoperative day, by verbal scale scoring every 4 h, by reporting techniques and analgesic drugs, and side effects. An audit of the nurses, was performed through a questionnaire in 1999. RESULTS: Four thousand sixty-eight patients were included in the audit over 5 years (1995 1999). Twelve objectives out of 15 were reached. Painful patients were less numerous, said themselves less painful, were more quickly relieved, and were satisfied by their management. The incidence of side effects was much lower than the figures of the literature. CONCLUSION: Pain control on mobilization, maintenance of a high quality continuous formation, and development of pain control process in the other surgical departments are the new goals in progress. PMID- 12747984 TI - [In-hospital cardiac arrest resuscitation: medical and paramedical theory skill assessment in an university hospital]. AB - OBJECTIVES: Theory knowledge and attitude assessment about in-hospital cardiac arrest (CA) basic life support by hospital staff. METHODS: We tested medical and paramedical working staff in a 450 beds university hospital with anonymous questionnaire based on training for basic life support. Questions were about presence of a formation before, clinical signs reached in front of unconscious patient, attitude in front of cardiac-arrest, practical experience with basic life support and ward emergency trolley. RESULTS: Five hundred and seventy one on 996 people answered to handed out questionnaires (57%): 158 from medical staff (Med group) and 413 from nurses "Pmed". Seventy one percent people from "Med" group and 64% from "Pmed" received one time at least training about cardiac arrest. Front of unconscious patient, no spontaneous breath was reached explicitly by 55% people from Med group and 19% from "Pmed" group and central pulse was reached explicitly by 70% people from "Med" group and 18% from "Pmed" group. Front of CA, 50% people from "Med" group released airway, 75% began ventilation and 86%, External Heart Compression (EHC) and 42% called for rescue. There were respectively 29, 47, 64 and 60% people from "Pmed" group. Eighty-one percent people from "Med" group thought they knew to do ventilation and 82% did it one time at least. Eighty-eight thought they knew to do EHC and 85% did it one time at least. They were respectively 67, 76, 73 and 78% people from Pmed group. Sixty-four per cent of Med group people know that there is emergency trolley in there department versus 89% for "Pmed" group. CONCLUSION: This study showed that theoretical knowledge of hospital staff about cardiac arrest diagnostic and management are insufficient from the point of view of national and international guidelines. Analysis is difficult because of weak response number and knowledge people overestimation. PMID- 12747986 TI - [Surgical emergencies at Libreville hospital center]. AB - OBJECTIVE: To estimate the waiting times of surgical emergencies and identify causes of delay. Study design. - Prospective study over 36 months. MATERIAL AND METHOD: From January 1996 to December 1998, surgical emergencies arriving to the Libreville Hospital Center have been classified in absolute emergencies (AE) and relative emergencies (RE). Five parameters have been measured: waiting before the first contact with the resident (T1); waiting time for making the decision of operation (T2); waiting time for the admission in the operating block (T3); waiting before the surgical intervention (T4); total duration of waiting between admission in the operating block and the beginning of the surgical intervention (TT). Concerning each one of those stages, the abnormal lengthening causes of time for surgical emergencies have been identified. RESULTS: During that period, 325 surgical emergencies have been received. The mean duration of surgical emergency management has been 504.3 +/- 613.7 min. Among those emergencies, 114 have been classified AE (35.1%) and 211 RE (64.9%) with mean durations of 421.2 +/- 347.0 min for AE and of 549.3 +/- 722.5 min for RE. Some delays in the management of emergencies have been found among 176 patients (54.2%). Their frequency has been 58.8% for AE and 51.7% for RE. The most current cause of delays has been the waiting of complementary medical tests results (44.4%), followed by difficulties in supplying (31.1%) and by technical or staff problems (24.1%). CONCLUSION: This study has permitted to show that the waiting times of surgical emergencies management are abnormally long in Libreville hospital center. There is an important frequency in delays as well for RE as for AE. Socio economic problems are very important. It seems necessary to organize surgical emergencies management in a specific structure and codify the prescription of complementary medical tests. PMID- 12747987 TI - [Which propofol target concentration for ASA III elderly patients for conscious sedation combined with regional anaesthesia?]. AB - OBJECTIVE: To determine the optimal propofol target concentration between 0.9-1.1 and 1.3 mg l(-1), for conscious sedation and amnesia using Diprifusor in ASA III patients over 60 years. STUDY DESIGN: Prospective study. PATIENTS AND METHODS: In ASA III patients over 60 years presented for elective vascular procedures under regional anaesthesia, sedation was induced with propofol TCI by increasing target concentrations from 0.9 to 1.3 mg l(-1) by 0.2 mg l(-1) steps up to a sedation score corresponding to light level (patient awakable with tactile stimulation). At baseline and each step, specific pictures were shown to the patient and clinical parameters and unwanted side effects occurrence were recorded. In PACU, memorisation of pictures and events was looked for. RESULTS: Conscious sedation was obtained in 100% of the patients at 0.9 mg l(-1), 94% at 1.1 and 78% at 1.3 mg l(-1). Amnesia was concentration-dependent and for each concentration was always more important for pictures than for events. Haemodynamic parameters did not change significantly. Incidents occurred in 12% of cases at 1.1 and 39.4% at 1.3 mg l(-1). DISCUSSION: None of those three concentrations was satisfying in 100% of cases for the three criterias (sedation < S2, amnesia and none side effects). These results suggest that propofol target concentration 0.9 microg ml( 1) could be used safely for sedation in elderly ASA III patients. Moreover, we have shown that amnesia for events requires higher propofol concentrations than amnesia for pictures during conscious sedation. PMID- 12747985 TI - [Out-of hospital management of elderly patients]. AB - OBJECTIVE: To evaluate prehospital management of elderly patients, agreement between prehospital and hospital diagnosis and to observe clinical course during hospitalization. TYPE OF STUDY: Retrospective study. PATIENTS AND METHOD: Out-of hospital patients of 65-year-old or more were included. Apart from demographic data, were collected: reasons for call, medicalization length, SAPS score, prehospital management, destination, prehospital and hospital diagnosis and patients evolution. Three groups were defined: G1 (65-74), G2 (75-84), G3 (> 84 year old). Statistical analysis was done by an Anova for quantitative data and by a Chi squared test for qualitative data. RESULTS: Two hundred and seventy-one patients were included (mean age 80 +/- 8 years, 43% of men). Eighty-two per cent of interventions were followed by a medicalized transport. Twelve per cent of patients died in the field. Forty-four per cent were hospitalised in intensive care unit, but patients of more than 84 year-old were significantly less often admitted in intensive care unit. There was no difference between the three groups in term of degree medicalization during transport. Eight per cent of patients required tracheal intubation in the field. Prehospital diagnoses were in agreement with reason for call in 61% of patients and with in-hospital diagnosis in 85% of patients. Fifty three per cent of patients came back home after hospitalisation. CONCLUSION: Analysis of elderly patient evolution after hospitalisation confirms the idea that the age should not influence the decision and the degree of prehospital medicalization. PMID- 12747988 TI - [Morphology and physiology of the blood-brain barrier]. AB - The blood-brain barrier (BBB) is a complex biological system that consists of endothelial cells, pericytes and astrocytes, which are involved in the induction and maintenance of its physiological and ultrastructural characteristics. The BBB plays a primordial role in isolating the cerebral parenchyma as well as in controlling brain homeostasis by its selective permeability to nutriments and other molecules flowing through the cerebral microcapillaries. A better knowledge of this system is crucial in order to improve the efficiency of brain penetration by drugs, and in order to prevent BBB opening, leading to brain edema, in physiopathological situations such as brain ischemia, trauma or inflammatory processes. PMID- 12747989 TI - [Osmotic cerebral oedema: the role of plasma osmolarity and blood brain barrier]. AB - There are five types of oedema: vasogenic, cytotoxic, interstitial, hyperemic and osmotic. The differences lie on the type and localization of the oedema, the state of the blood-brain barrier (BBB) and the pathological context. Under physiological conditions, the osmolarity of extra cellular fluids (ECFs) is equal on both sides of the BBB. However, the pathophysiological variations of circulating osmolarity (including acute hyponatremia and hypernatremia) do not affect, at the same time, the osmolarity of cerebral ECFs. This situation generates an osmotic gradient on either side of the BBB. The latter, if intact, behaves like a semi-permeable membrane allowing water transport according to the osmotic laws. Depending on its direction, water movement could induce cerebral liquid inflation (i.e. osmotic oedema) or cerebral dehydration. In case of osmotic insult, cerebral cell modify their active osmotic molecular contents in order to limit volume variation. There are two types of osmoactive molecules, organic (i.e. ideogenic osmoles: amino acids, polyols and trimethylamines) and non organic (i.e. electrolytes). In the event of plasma hypotonicity, cerebral cells expel active osmotic molecules to reduce the osmotic gradient and water movement thereby reducing edema. The opposite reaction is observed in the case of hypertonic insult. This cerebral osmoregulation becomes more effective, the slower the osmotic disorder. It explains, for example, why patients with chronic and severe hyponatremia could be asymptomatic. Severe osmotic oedema is observed mainly in water intoxication, acute hyponatremia or too rapid reduction of hyperosmolarity. However, osmotic oedema is not limited to extreme clinical circumstances. Hyponatremia, even modest, could modify cerebral blood volume and impair osmoregulation. Generally these minor modifications do not affect normal brain tissue. In the presence of cerebral lesion, osmoregulation operates only in areas of preserved BBB. The pathological zones are therefore exposed to osmotic oedema (even in cases of moderate hyponatremia) with deterioration of both clinical status and intracranial pressure. This authentic phenomenon could be insidious and difficult to differentiate from osmotic central oedema. Hyponatremia constitutes an authentic secondary cerebral insult of systemic origin, an entity clearly identified by experimental studies to justify the choice between crystalloids and colloids in neuroanaesthesia and neurointensive care. These studies have revealed an increase in water content in normal brain tissues after administration of hypotonic solutions. The increase in plasma osmolarity as a treatment modality using mannitol or hypertonic saline is based on the same concepts. The most remote indication is the occurrence of a reactive mydriasis in the context of trauma for example. More recently, therapeutic hypernatremia has been proposed to control intracranial hypertension. PMID- 12747990 TI - [Brain oedema following blood-brain barrier disruption: mechanisms and diagnosis]. AB - Brain oedema following blood-brain barrier (BBB) disruption, or vasogenic oedema, is present in most cases of brain oedema. According to the Starling's law, water, ions and plasma proteins cross the BBB toward the interstitium if the driving forces for transmural bulk flow are excessive (mechanical origin) and/or if the BBB permeability is enhanced (chemical origin). Both mechanisms coexist in most cases. Excessive elevation of the gradient of hydrostatic pressure with lost of cerebral autoregulation has been proved in ischaemia/reperfusion and trauma, and suggested in acute mountain sickness and eclampsia. The BBB permeability can be enhanced by immediate (chemical mediators) or delayed (cellular infiltration) inflammatory response, or by alteration of the membrane integrity. This later can be transient (hyperosmolar BBB disruption), or permanent by activation of matrix metalloproteinase or by neovascularization with BBB breakdown. The reference method for the diagnosis of vasogenic oedema is the MRI diffusion-weighted imaging. PMID- 12747991 TI - [Anaesthesia for patients with intracranial hypertension due to cerebral oedema]. AB - The main objective for anaesthesia in patients with intracranial hypertension (ICH) is to maintain the cerebral perfusion pressure (CPP). Before the operation, the assessment of the level of intracranial pressure relies on the Glasgow coma score and the signs of ICH on the CT-scan. In the perioperative period, repeated transcranial Doppler examinations may help in determining the adequate CPP. Haemodynamic and respiratory complications are common after subarachnoid haemorrhage or head injury. Careful preoperative screening of the cardiovascular and respiratory system is mandatory before anaesthesia. There is no recommended anaesthetic technique for patients with ICH. Nitrous oxide should be avoided in patients with severe ICH or during emergency surgery. Theoretically, intravenous anaesthesia is a better choice than inhalation anesthesia because of the cerebral vasodilatation induced by inhalation agents. In the most severe cases thiopental is the only anaesthetic agent to consider. Treatment of hypovolaemia with fluid loading and the early use of vasoactive agents can be recommended to maintain CPP. Before intracranial surgery, large doses of mannitol have been demonstrated to improve neurological recovery in brain injured patients. The urinary losses due to the infusion of mannitol should be replaced with isotonic saline. Emergence and extubation are best performed in the intensive care unit under close systemic and cerebral haemodynamic control. PMID- 12747992 TI - [Automated external defibrillator use during cardiopulmonary resuscitation during flight]. AB - Automated External Defibrillator (AED) during cardiopulmonary resuscitation should reduce mortality rate after out-of-hospital cardiac arrest. We report a case of defibrillation with AED during flight in a patient suffering cardiac arrest complicating an acute myocardial infarction. Two hours before landing, a 56-years-old man presented sudden cardiac arrest. Flight attendants performed basic cardiac life support, including AED. Five shocks were delivered. After landing, acute myocardial infarction was diagnosed and treated by prehospital thrombolysis and angioplasty with favorable outcome. AED is a crucial link of the chain of survival, especially where advance cardiac live support cannot be performed, like during flight. Despite an increasing AED availability, survival after cardiac arrest during flight remains exceptional. PMID- 12747993 TI - [Paraplegia and pregnancy: anaesthesic management]. AB - Pregnancy is a rare occurrence in patient suffering from spinal cord lesion. The authors report the case of labour and delivery in a paraplegic patient who was suffering from lesion located T6 level. In early labour epidural analgesia was maintained and the vaginal delivery was successful without associated fluctuation of blood pressure or other signs of autonomic hyperreflexia. The epidural catheter was maintained for 48 h post-partum. The main risks and anaesthesic management of pregnancy in paraplegic patients are discussed. PMID- 12747994 TI - [Neuromuscular blockade monitoring at the corrugator supercilii and ocular myasthenia gravis]. AB - A 67-year-old patient suffering from an ocular myasthenia gravis was scheduled for an elective ENT surgery. General anaesthesia was induced intravenously. Neuromuscular responses after train-of-four stimulation were normal at both the adductor pollicis (T(4)/T(1) = 1) and the corrugator supercilii (4 visual responses). Then cisatracurium (0,15 mg kg(-1)) was administered to allow tracheal intubation. The laryngoscopy attempted 45 s after cisatracurium injection (no response at the supercilii, T(1)/T(0) = 1 at the adductor pollicis) was unsuccessful because of closing and moving vocal cords. The second attempt was successful 4 min after cisatracurium injection (no response at the corrugator supercilii, T(1)/T(0) = 0.05 at the adductor pollicis). Residual neuromuscular blockade was antagonized at the end of surgery (1 h long) allowing an uneventful extubation. We concluded that monitoring neuromuscular blockade at the corrugator supercilii to assess the intubating conditions is not recommended in a case of ocular myasthenia gravis. PMID- 12747995 TI - [Prevention of the violence in hospital environment]. PMID- 12747996 TI - [About amniotic fluid embolism]. PMID- 12747997 TI - [Impact of quality system on mortality in anaesthesia: experience of Monastir university hospital]. PMID- 12747998 TI - [When difficult intubation becomes impossible!]. PMID- 12747999 TI - [Use of the lumbar facet joint L4-L5 for spinal anaesthesia in the elderly]. PMID- 12748000 TI - [The decision to cease treatment at the end of life: the hidden face of disease]. PMID- 12748005 TI - I'm not going to take it anymore. PMID- 12748001 TI - Teratology in the 20th century: environmental causes of congenital malformations in humans and how they were established. PMID- 12748006 TI - Principles of treatment of the pineal region tumors. AB - BACKGROUND: A pineal region tumor is an uncommon deep-seated, heterogeneous group of mass lesions of the brain, and the management strategy of any types of these tumors remains controversial. It is the purpose of this communication to present results of our experience in treating patients with these neoplasms. METHODS: From 1976 to 1999 about 700 patients with tumors of the pineal region and posterior third ventricle were managed at the Burdenko Neurosurgery Institute. In more than 330 cases the tumor was removed. In this paper we present results of 287 patients with histologically verified pineal region tumors for the period from 1976 to 1999. All of them had verified tumor histology, excluding only five cases in which stereotactic biopsy procedures were uninformative. There are four main groups of tumors: the germ cell tumors-87 (31%); the pineal parenchymal tumors-75 (27%); the glial tumors-77 (27%); and miscellaneous-43 (15%). There were 255 surgical procedures for tumor removal performed in 244 and stereotactically guided biopsies in 61 patients, 168 (58%) with obstructive hydrocephalus who underwent cerebrospinal fluid shunting. Radiation therapy was administered in 145 (51%) and chemotherapy in 16 patients. Follow-up data were collected in 209 cases (80%). Survival was calculated with the Kaplan-Meier product limit method. RESULTS: The occipital transtentorial and infratentorial supracerebellar approaches were the most commonly used on our series-138 (54%) and 87 (34%), respectively. In the remaining cases, subchoroidal approach, approach through the fourth ventricle, and the combined approaches were used. A total tumor removal was achieved in 148 operations (58%), subtotal in 74 (29%) and partial in 33 (13%). The projected 5-year and 10-year survival rates for patients with malignant pineal tumors, who received irradiation after tumor resection or underwent radiation therapy alone, were: 95% and 88% for pure germinomas, 80% and 50% for high grade gliomas, 44% and 0% for malignant pineal parenchymal tumors, and 20% and 0% for malignant germ cell tumors, respectively. CONCLUSIONS: Benign pineal tumors should be cured with surgery alone. Malignant tumors should be treated with aggressive resection followed with irradiation and chemotherapy. Pure germinomas, which are exquisitely radiosensitive, can be cured by conventional radiation therapy alone. PMID- 12748007 TI - Importance of neuropsychological evaluation after surgery in patients with unruptured cerebral aneurysms. AB - BACKGROUND: We evaluated neuropsychological function before and after surgery in patients with unruptured cerebral aneurysms. METHODS: Neuropsychological functions in 43 patients with unruptured cerebral aneurysms were evaluated before and 1 month after surgery. The neuropsychological examination included the Mini Mental State examination, "Kana-hiroi" test, Kohs Block Design test, and Miyake's Memory test. Then, if scores of even a single test were decreased 1 month after surgery, the tests were performed again 5 months later. In 24 of the 43 subjects, cerebral blood flow (CBF) was measured before and 1 month after surgery by single photon emission tomography. RESULTS: The outcome in all patients was evaluated as good according to the Glasgow Outcome Score (GOS). In 17 (40%) of the 43 patients, neuropsychological function had deteriorated 1 month after surgery. The most sensitive test applied was the Miyake's Memory test. Of 14 patients with neuropsychological deterioration 1 month after surgery, 6 showed complete recovery, 5 showed partial recovery, and 3 still showed cognitive deterioration 6 months after surgery. Patients over 65 years old, those with anterior communicating artery aneurysms, those operated by interhemispheric approach, or those with systemic diseases showed a greater tendency toward a decline in postoperative neuropsychological function than the other patients. The postoperative CBF and vascular response in the frontal lobe of affected-side was decreased in cases showing cognitive deterioration. CONCLUSIONS: These results suggested that the neuropsychological outcomes after surgery for unruptured cerebral aneurysms were not satisfactory. PMID- 12748009 TI - MRI-guided stereotactic biopsy in the diagnosis of glioma: comparison of biopsy and surgical resection specimen. AB - BACKGROUND: Although there has been a dramatic increase in the accessibility and utilization of high-resolution MRI techniques for the evaluation of brain tumors, there is currently only a single report comparing stereotactic brain biopsy specimen to subsequent resection specimen exclusively in the management of gliomas. METHODS: The diagnoses in 43 cases of astrocytic brain tumors were derived using MRI-guided stereotactic biopsy followed by open resection of the lesion. The histologic diagnoses yielded by biopsy were compared with subsequent histologic diagnosis after open tumor resection. All biopsies and histologic diagnoses were made by the same surgeon and pathologist, respectively. RESULTS: In 23 patients undergoing resection <60 days after biopsy, the biopsy diagnosis was consistent with resection diagnosis in 18 cases (79%) and led to the correct treatment in 22 cases (96%). Recurrent glioblastoma multiforme (GBM) was undergraded as anaplastic astrocytoma in 4 patients. GBM was misdiagnosed as radiation necrosis in 1 patient. MR-nonenhancing lesions [10/10 (100%)] yielded histology that correlated with subsequent craniotomy, while only 8/13 (61%) MR enhancing lesions yielded histology that was consistent with that at craniotomy (p < 0.05). In 20 patients undergoing resection because of radiologic tumor progression (mean 7 months after biopsy), 6/6 (100%) biopsy diagnoses of a specific glioma grade correlated with resection diagnosis, while only 6/14 (43%) biopsy diagnoses of radiation effect correlated with resection diagnosis (p < 0.01). CONCLUSION: MRI-guided stereotactic brain biopsy specimen accurately represents the grade of the larger glioma mass sufficiently to guide subsequent therapy. Enhancement on MR may be a negative prognostic indicator of biopsy accuracy. PMID- 12748011 TI - Surgery combined with radiosurgery of large acoustic neuromas. AB - The treatment of acoustic neuromas has been improved by advancements in microsurgical techniques and in radiosurgery. To further elucidate the degree of clinical improvement, we evaluated the treatment results of a combination of surgery and radiosurgery for large acoustic neuromas. METHODS: From January 1994 through December 2000, we treated 14 patients with large acoustic neuromas using a combination of surgery and radiosurgery. Of these, 8 were male and 6 were female patients, with an average age of 47 years (range, 18-64). The average maximum diameter of the tumor was 42 mm (range, 30-58 mm). All patients underwent operations using the retrosigmoid approach, and one patient was retreated using the transpetrosal transtentorial approach. The tumors were removed subtotally in thirteen patients and partially in one who had a very large hypervascular acoustic neuroma. There were no mortality and no surgical complications, such as hemorrhage or CSF leakage. Postoperative facial palsy was avoided in 10 patients (71%). Radiosurgery was performed 1 to 6 months (mean, 2.9 months) after surgery. At the time of radiosurgery, the treatment size (mean diameter) became 19.2 mm (range, 9.8-36.1 mm). The average tumor marginal dose was 12.1Gy (range, from 10 14 Gy). The mean follow-up period was 32 months after radiosurgery. RESULTS: The tumor size decreased in 6 patients, unchanged in 5 patients, and increased in 3 patients. Only 1 patient (7%) with extra large tumor needed surgical resection 1 year after radiosurgery. Excellent facial nerve function (House & Brackmann Grade I or II) was preserved in 12 patients (85.7%) in the final follow-up. CONCLUSIONS: In the case of large acoustic neuromas, subtotal removal and subsequent radiosurgery is one option for maintaining cranial nerve function and long-term tumor growth control. PMID- 12748013 TI - Usefulness of multiple-site venous sampling in the treatment of adrenocorticotropic hormone-producing pituitary adenomas. AB - BACKGROUND: While venous sampling for adrenocorticotropic hormone (ACTH) may be the most accurate way to establish a diagnosis of Cushing's disease, its usefulness for lateralization of adenomas is limited even in combination with simultaneous bilateral sampling methods. Therefore, to increase accuracy in predicting the lateralization of adenomas, we performed a trial in which we carried out simultaneous venous sampling from multiple sites of the cavernous sinus (CS) and inferior petrosal sinus (IPS). Here we report on the usefulness of our method in the correct tumor lateralization in patients with Cushing's disease. METHODS: Eighteen patients with Cushing's disease underwent simultaneous bilateral ACTH sampling. The samples were obtained from the anterior, middle, and posterior CS and the IPS. CS sampling after stimulation with corticotropin releasing hormone (CRH) was also performed. The central-to-peripheral ACTH ratio (c/p ratio) was calculated using sampling data from each site; the lateralizing gradients (right versus left) were calculated using the c/p ratio. Instead of the conventional method where the lateralization gradient is based on sampling data from a single site, we used the distribution of the c/p ratio determined from multiple-site data. RESULTS: There was no significant difference in the mean lateralization gradient obtained from each set of sampling data (p > 0.05). With the conventional method, correct tumor lateralization was obtained in 50% of tumors in the IPS; 72.2 to 77.8% of tumors in the CS; and 77.8% of tumors in the middle CS after CRH. Our method using ACTH contour analysis based on multiple sites produced correct results in all 18 of the patients examined. The difference in correct calls between the conventional method and our method was significant (p < 0.05). CONCLUSIONS: Multiple-site sampling of ACTH is valuable for lateralizing the adenoma in patients with Cushing's disease; it produces more correct results than does single venous sampling. PMID- 12748015 TI - Vertebral artery complications in anterior approaches to the cervical spine: report of two cases and review of literature. AB - BACKGROUND: Cases of lesions to either the carotid artery or the vertebral artery in anterior approaches to the cervical spine are rarely found in medical literature. Two cases of vertebral artery injury in anterior approaches as well as a review of the pertinent literature are presented. In cases of arterial injury, appropriate management strategies are necessary to avoid or minimize harm to the patient. CASE REPORTS: In the first case, the vertebral artery was injured during decompression of a cervical spinal stenosis while drilling the neuroforamen. Local compression provided sufficient control of hemorrhage. Nevertheless, rebleeding from a pseudoaneurysm occurred 2 days later. After removal of the hematoma, the pseudoaneurysm was treated successfully with coils by an endovascular approach. In the second case, misplacement of one screw in screw-fixation of a type II odontoid fracture caused a pseudoaneurysm of the vertebral artery. This led to a fatal subarachnoid hemorrhage 4 days later. CONCLUSIONS: In ventral approaches to the cervical spine, precise preoperative planning and a detailed knowledge of the surgical anatomy are mandatory. In cases of injury to the vertebral arteries, direct surgical repair is most appropriate to prevent complications arising from fistulas, late-onset hemorrhages, pseudoaneurysms, thrombosis, and emboli. Alternatively, endovascular techniques or even clipping or ligation of the affected artery should be considered. PMID- 12748017 TI - Evaluation of intra-aneurysmal blood flow by digital subtraction angiography: blood flow change after coil embolization. AB - BACKGROUND: The natural course of cerebral aneurysms is related to many factors, and it is very important that intra-aneurysmal blood flow is considered. Our group developed a method that allowed the simultaneous evaluation of blood flow in human cerebral aneurysms using digital subtraction angiography (DSA) with no special devices. The intra-aneurysmal blood flow measurement would also be very useful for coil embolization. Since the Guglielmi detachable coil (GDC) was developed, many patients with cerebral aneurysm have been treated with GDC, but coil compaction has sometimes caused a problem after the coil embolization of a cerebral aneurysm. We believed that an intra-aneurysmal flow measurement would suggest the final result of embolization during the procedure. METHODS: We performed DSA to examine 17 aneurysms in 17 patients. The video signal of serial DSA images was stored on a personal computer, and time-density curves were obtained for each individual pixel. The formula, determined by a two-exponential model, was fitted to the time-density curve 1000 times by least square approximation for each individual pixel. We indirectly substituted the coefficient of the flow-in curve for the blood flow. We were therefore able to display the distribution of intra-aneurysmal blood flow in color. We could compare the blood flow in each portion of the cerebral aneurysm and parent artery during coil embolization. RESULTS: The blood flow k(a) in a small aneurysm was faster than that in a large aneurysm, and it slowed in accordance with the coil embolization. The blood flow in a large aneurysm was sometimes accelerated by incomplete coil embolization. CONCLUSION: We can detect the flow distribution in cerebral aneurysms and the flow change during coil embolization, using existing equipment. Our method would be useful in elucidating the natural history of cerebral aneurysms, treating cerebral aneurysms with coils, and following patients after treatment. PMID- 12748019 TI - Multiple cerebral abscesses because of Listeria monocytogenes: three case reports and a literature review of supratentorial listerial brain abscess(es). AB - BACKGROUND: Central nervous system involvement often follows bacteremia because of Listeria monocytogenes. Meningitis is clinically the most common manifestation, while brain abscess occurs in about 1% of patients. Brain abscess is usually solitary but in recent years, probably in part because of the availability of computerized tomography and magnetic resonance imaging, several reports have described two or more separate supratentorial abscesses. METHODS: We have described three patients with listerial brain abscesses and reviewed the North American and European literature of brain abscess(es) because of L. monocytogenes through December 2001. We have evaluated the role of underlying diseases and therapeutic immunosuppression on the development of solitary or greater than one brain abscess. RESULTS: In contrast to meningitis, where immunosuppression does not predispose either to disease incidence or to higher mortality, patients with solitary and particularly those with more than one supratentorial abscess usually are immunosuppressed either by disease or by therapy. Corticosteroids in particular are significant predisposing factors, especially in those patients with two or more brain abscesses. Mortality resulting from listerial brain abscess, whether solitary or multiple, is nearly three times higher than nonlisterial brain abscess, probably in part because of both underlying diseases and immunosuppressive therapy. CONCLUSIONS: Therapy with high-dose ampicillin in combination with gentamicin appear to be the drugs of choice, followed by trimethoprim/sufamethoxazole and vancomycin. In general, antimicrobial therapy appears to be satisfactory treatment without surgical intervention. PMID- 12748020 TI - A modified technique to treat chronic and subacute subdural hematoma: technical note. AB - BACKGROUND: We present a patient on warfarin in whom a drainage port system was attached to the skull, successfully draining a subacute subdural hematoma. CASE DESCRIPTION: An elderly male presented to our institution with right hemiparesis a week following a motor vehicle accident. He was on warfarin for recurrent pulmonary emboli and suffered from severe coronary artery disease. Physical examination demonstrated a grade 3/5 hemiparesis and a computerized tomography (CT) scan confirmed the diagnosis of subacute subdural hematoma. He underwent twist drill craniostomy and attachment of the subdural evacuating port system. Recovery in this patient was dramatic. CONCLUSION: The subdural evacuating port system (SEPS) permits the neurosurgeon to drain subacute or chronic hematomas by a method that is minimally invasive, simple, and safe. The SEPS appears to promote brain expansion without the potential biohazards of other standard techniques. PMID- 12748022 TI - Patients' bill of rights or trial lawyers' bill of delights. PMID- 12748023 TI - "United we stand...". PMID- 12748024 TI - Why everyone picks on doctors. PMID- 12748025 TI - Re: Ausman JI. The death of cerebral aneurysm surgery. Surg Neurol 2001:56;348. PMID- 12748026 TI - Managed care organizations and access to mental health specialists: it's a jungle out there. PMID- 12748027 TI - Managed care organizational complexity and access to high-quality mental health services: perspective of U.S. primary care physicians. AB - This analysis addresses the relationship between perceived access to high-quality specialty mental health and medical services and 2 aspects of managed care organizational complexity at the practice level: 1) gatekeeper requirements for specialty services, and 2) managing multiple contracts. Cross-sectional analysis of a national telephone survey of 7,197 primary care physicians (PCPs) was performed. Access was defined as high-quality specialty services being always or almost always available to the PCP's patients when medically necessary. PCPs rated access to high-quality outpatient specialty mental health services as much lower than that of specialty medical services (28%; 95% confidence interval [CI], 27-29 versus 81%; 95% CI, 80-82). After adjustment for physician, practice, and managed care factors (multiple logistic regression analysis), perceived access to high-quality outpatient mental health services was lowest for practices with the largest number of managed care contracts and when a physician's practice was a "mixed model" with regard to the gatekeeper function. Perceived access to high quality specialty medical services was not as strongly associated with these practice characteristics. PCPs who interact with a large number of managed care plans and different administrative models may have the most difficulty in obtaining high-quality mental health services for their patients. PMID- 12748028 TI - Improving primary care treatment of depression among patients with diabetes mellitus: the design of the pathways study. AB - This paper describes the methodology of a population based study of primary care patients with diabetes mellitus enrolled in a health maintenance organization. The first goal was to determine the prevalence and impact of depression in patients with diabetes. The second goal was to randomize approximately 300 patients with diabetes and major depression and/or dysthymia in a trial to test the effectiveness of a collaborative care intervention in improving quality of care and health outcomes among patients with diabetes and depression. PMID- 12748029 TI - The pilot study of a telephone disease management program for depression. AB - Most depressed patients are seen and treated exclusively by primary care clinicians. However, primary care patients with depression are often not adequately treated. The aims of this pilot study were to measure the impact of a telephone disease management program on patient outcome and clinician adherence to practice guidelines, measure the relationship of clinician adherence to patient outcome, and explore the measurement of patient adherence to clinician recommendations and its impact on patient outcomes. Thirty-five primary care practices in the University of Pennsylvania Health System were randomized to telephone disease management (TDM) or "usual care" (UC). All patients received a baseline and a 16-week follow-up clinical evaluation performed over the telephone. Those from TDM practices also received follow-up contact at least every 3 weeks, with formal evaluations at weeks 6 and 12. These interval contacts were designed to facilitate patient and clinician adherence to a treatment algorithm based on the Agency for Health Research and Quality (AHRQ) practice guidelines. Depressive symptoms evaluated with the Community Epidemiologic Survey of Depression (CES-D) scale as well as guideline adherence were the primary outcome measures. Sixty-one patients were enrolled in this pilot project. The overall effect for CES-D scores over time was significant, (P <.001), indicating that those participating in the trial (both TDM and UC groups) showed significant improvement. The interaction between intervention condition and time was also significant (P <.05), indicating that TDM patients improved significantly more over time than did UC patients. A greater proportion of TDM patients had CES-D scores <16 by Week 16 (66.7 versus 33.3%; chi(2), P <.05). The improvement in depression outcome for the TDM group was related to its impact on improving clinician adherence to depression treatment algorithms. The TDM pilot did not show a statistically significant effect on improving patient adherence to clinician recommendations, however. This preliminary data suggests that TDM for depression improves both clinician guideline adherence and patient outcomes in the acute phase of depression. The effect on patient outcome is at least partially explained by the effect of TDM on clinician adherence to depression treatment algorithms. PMID- 12748030 TI - Psychiatric symptoms, impaired function, and medical care costs in an HMO setting. AB - More information is needed regarding the medical care utilization and costs of individuals who report depressed mood, persistent anxiety, brief anxiety, panic, and trouble controlling violent behavior. We present findings from a 1-year prospective follow-up study of a stratified random sample of adult HMO enrollees (N = 10,377) originally interviewed by telephone. A strong association was observed between these psychiatric symptoms, associated impaired function, and general medical care costs during the year following the interview. After controlling for age, gender, race, medical conditions, and smoking, the mean costs of general medical care were $1,948 for respondents who reported none of the psychiatric symptoms or impaired function: $3,006 for respondents with all 5 symptoms but no impaired function; and $3,906 for those with all 5 symptoms and pervasive functional impairment. Persistent anxiety and depressed mood had the greatest impact on total general medical costs, while impaired function was associated with increased likelihood of hospital admission and emergency room use. We conclude that depressed mood, persistent anxiety, and related impaired function are associated with substantial increases in the use and cost of general medical care. PMID- 12748031 TI - Major depressive disorder in the general hospital: adaptation of clinical practice guidelines. AB - Major Depressive Disorder is particularly frequent among physically ill inpatients. Despite the considerable human burden and financial costs, Major Depressive Disorder remains under-detected and under-treated. To improve this situation, clinical practice guidelines for the management of Major Depressive Disorder were developed for patients in the general hospital. They were adapted from existing good quality guidelines. A literature search has been conducted to identify guidelines and systematic reviews about the management of Major Depressive Disorder. The quality of the existing guidelines was evaluated by means of the AGREE instrument (Appraisal of Guidelines for Research and Evaluation). Complementary literature searches were necessary to answer questions such as "depression and physical illness" or "antidepressants and somatic medication". The guidelines were discussed by a multidisciplinary internal panel. The final version was reviewed by an external panel. This paper presents the development process and a summary of these guidelines for the management of Major Depressive Disorder. The adaptation of good quality guidelines to local needs requires much time, effort and skills. Easier ways for the adaptation and use of high quality guidelines at the local level may result from better coordination, organization and updating of guidelines at a national or supranational level. PMID- 12748032 TI - The role of cognitive impairment in desire for hastened death: a study of patients with advanced AIDS. AB - To examine the relationship between cognitive impairment and desire for death in patients with advanced AIDS we evaluated 128 patients with advanced AIDS consecutively admitted to three long-term care facilities. Participants completed a clinical assessment that included a self-report measure of desire for hastened death in the medically ill (Schedule of Attitudes toward Hastened Death), three measures of cognitive functioning designed to screen for cognitive impairment (Dementia Rating Scale, Mini-Mental State Exam, HIV Dementia Scale), and other measures of physical and psychological functioning. Participants who were classified as cognitively impaired obtained significantly higher scores on the measure of desire for death than did patients without cognitive impairment. This modest association between cognitive impairment and desire for death remained significant even after controlling for the impact of depression on desire for death. Specific aspect of cognitive functioning such as memory and psychomotor coordination appeared to be more salient than executive functioning or abstract reasoning. Cognitive impairment appears to have a modest, but significant impact on patients' desire for hastened death. Aggressive treatment of cognitive symptoms in the terminally ill is necessary in order to disentangle the various factors that may drive end-of-life treatment decisions. PMID- 12748033 TI - Assessing risk for major depression on patients selected for percutaneous transluminal coronary angioplasty: is it a worthwhile venture? AB - We hypothesized that a prior history of a major depressive disorder would not compromise the efficacy of percutaneous transluminal coronary angioplasty (PTCA), a coronary revascularization procedure, in improving quality of life and health status when comparing patients with no previous history. To determine the utility of screening for risk for depression in heart patients scheduled for PTCA, 190 patients were administered a two-item depressive disorders screener prior to PTCA and the SF-36 and Seattle Angina Questionnaire prior to and 6 months post procedure. Results reveal that while those with no prior history of depression had statistically better quality of life and health status outcomes than those with a probable past depression, (P <.05), the clinically meaningful differences as determined by effect size scores showed that those susceptible to recurrent depression benefited from PTCA as well as, and on some measures better than those with no previous history. Prescreening patients at probable risk for a depression is not a valid or helpful enterprise if the purpose is to develop intervention strategies for improving outcome post PTCA. Screening out patients based on history of depression may in fact lead to an inequitable allocation of resources and have no major benefit in enhancing quality of life and improving health status. PMID- 12748034 TI - Erectile dysfunction: prevalence and relationship to depression, alcohol abuse and panic disorder. AB - We studied the prevalence of erectile dysfunction in a sample of 829 Nigerian men, using the International Index of Erectile Function questionnaire. These men were also screened for depression, alcohol abuse and panic disorder using the Patient Health Questionnaire. The prevalence of erectile dysfunction (ED) was 36% in men 30 years and below, 31% in those 31 to 40, 46% in those 41 to 50, and 58% in those 51 to 60. Among those men identified as having ED, 10% were depressed, 10.3% had alcohol abuse while 0.6% had panic disorder. Using a multiple linear regression model, age and depression were found to be good predictors of erectile dysfunction but not alcohol abuse and panic disorder. We suggest that ED may be much more common than is being reported. The use of simple questionnaires by doctors, especially those working in medical and surgical clinics and those in primary health care centers, could help in detecting more cases for whom the negative life impact of ED could be minimized. PMID- 12748035 TI - Do women who screen positive for mental disorders in primary care have lower mammography rates? AB - Disparities in mammography rates have been documented for underserved populations, yet no data are available for women with mental illness in primary care settings. We analyzed data on mammography rates for 526 women age 40-70 who were new patients and completed the Primary Care Evaluation of Mental Disorders (PRIME-MD). There were no significant differences in mammography rates among women who screened negative and positive for any mental illness (56% and 53%, respectively). Screening for mental disorders in primary care does not appear to identify women at risk for nonreceipt of mammography. PMID- 12748036 TI - Injured temporomandibular joint associated with fluoxetine-monotherapy-induced repeated yawning. PMID- 12748037 TI - Impact of exposure duration by low molecular weight compounds on interferon-gamma and interleukin-4 mRNA expression and production in the draining lymph nodes of mice. AB - The local lymph node assay (LLNA) is used to identify allergens by means of dermal exposure. For hazard identification, besides identification also the distinction between contact and respiratory allergens is of importance. We have previously shown that a modified LLNA can be used to identify respiratory allergens, on the basis of Con A induced IL-4 production. Here we show a good qualitative correlation between mRNA expression and production of IFN-gamma and IL-4. This suggests that distinction between contact and respiratory allergens may also be studied at the mRNA expression level. Secondly, another assay, similar to the modified LLNA but differing in the duration and the number of allergen applications as well as in the ex vivo culture conditions, here denoted as 'longer' assay, has been reported to be able to identify contact allergens, on the basis of (spontaneous) IFN-gamma production. In the present study we have compared these assays. Similar to our previous findings, in the modified LLNA exposure to the respiratory allergen trimellitic anhydride (TMA) resulted in a approximately 10-fold higher Con A induced IL-4 production compared with the contact allergen dinitrochlorobenzene (DNCB), while exposure to both allergens resulted in a similar Con A induced IFN-gamma production. In the 'longer' assay, TMA exposure resulted in Con A induced IL-4 production whereas DNCB exposure did not. Importantly, only a 2-fold higher spontaneous IFN-gamma production was induced by DNCB compared with TMA, the difference being not statistically significant. Thus, although the 'longer' assay indeed showed a somewhat higher IFN-gamma induction by DNCB compared with TMA, the magnitude and robustness of this effect question its applicability. These results favor the modified LLNA since it is shorter, and combines identification of allergens (by cell proliferation) with identification of respiratory allergens (by IL-4 production). Compounds that induce cell proliferation with a low concomitant IL-4 production may thus be identified as contact allergens, although the need to positively identity such allergens remain. PMID- 12748038 TI - Examining the relationship between impaired host resistance and altered immune function in mice treated with TCDD. AB - Exposure to TCDD suppresses the immune response to numerous antigens, including bacterial and viral pathogens. Although we administer a non-lethal infection with influenza A virus, we often observe significant mortality in TCDD-treated animals. With the goal of identifying which TCDD-induced defects impair host resistance, we conducted a dose response study to examine whether alteration of particular immunological endpoints could be correlated with mortality. C57Bl/6 mice were treated with vehicle control, or 1, 2.5, 5, 7.5 or 10 microg/kg TCDD 1 day prior to intranasal (i.n.) infection with influenza virus. Survival was monitored for 9 days, when remaining mice were sacrificed and multiple endpoints evaluated. Lymphocyte migration to the lung and the production of virus-specific IgG2a, IgG1, and IgG2b antibodies were significantly diminished, even at the lower doses. IgA was enhanced in all groups treated with TCDD. In contrast, T cell expansion in the lymph node, and the production of IFNgamma and IL-12 were relatively resistant to suppression. Treatment with TCDD also enhanced pulmonary neutrophilia in infected mice. These results suggest that decreased antibody production and hyperinflammation may contribute to the death of TCDD-treated mice, and underscore the importance of evaluating numerous endpoints before concluding that a chemical is or is not immunotoxic. PMID- 12748039 TI - Tritiated thymidine incorporation does not enhance sensitivity of the popliteal lymph node assay. AB - The popliteal lymph node (PLN) assay has been proposed as a tool to predict drugs and chemicals with the potential to induce systemic autoimmune reactions in man. In this assay, weight and cellularity indices typically are the measured endpoints. The present study was conducted to test whether incorporation of tritiated thymidine could improve sensitivity of the PLN assay. Male and female Balb/c mice were injected with 20 microCi of [3H]-methyl-thymidine intravenously 7 days after receiving 0.5, 1 or 2 mg of diphenylhydantoin, streptozotocin, sulfamethoxazole, ofloxacin, phenobarbital, or metformin intradermally. Results obtained with incorporation of tritiated thymidine were compared to weight indices. No consistent or marked differences in these endpoints were noted whatever the compound used. This study shows that incorporation of tritiated thymidine does not improve sensitivity of the PLN assay. PMID- 12748040 TI - Effects of low concentrations of cadmium on immunoglobulin E production by human B lymphocytes in vitro. AB - Exposure to cadmium (Cd) can cause a variety of biological effects including alterations of immune responses in animals and humans. Both immunosuppression and immunoenhancement have been reported. The present study was aimed at investigating the consequences of exposure to Cd on the human immunoglobulin (Ig) E synthesis, using purified peripheral blood B lymphocytes and IL-4 and anti human CD40 monoclonal antibody (a-CD40 mAb) as stimuli. Low concentrations of Cd (0.1-10 microM) markedly inhibited production of IgE in a concentration-dependent manner. IgG production, in contrast to IgE, showed a tendency towards being enhanced by Cd, although with a certain individual variability; IgM production was not affected. Cd failed to alter immediate surface expression of the activation markers CD69 and CD23 indicating that early activation events were not impaired. However, the portion of activated B cells was diminished by Cd after stimulation for more than 24 h, paralleled by a concomitant decrease in viability and a subsequent reduction in proliferation. These data suggest that the mechanism of Cd action on activated B cells involved pathways that interrupted an effectively initiated cell activation and induced a cytotoxic signal. Results from this study thus provide further evidence for and new information on the immunotoxic and immunomodulatory effects of Cd on human immune responses. PMID- 12748041 TI - Species differences in the structure and function of the immune system. AB - With the recent publication of regulatory guidelines from both the FDA and the CPMP addressing the investigation of immunotoxicity of new chemical entities has come the requisite increased application of immunotoxicology protocols. Importantly, the fulfillment of these protocols may require the use of different species, and while in many cases information concerning the structure and function of the immune system can be readily translated across species, there are numerous and significant species differences that need to be considered. In some cases, the generation of meaningful immunotoxicology data can be adversely affected by the choice of a species that does not adequately share the immune function of concern with man. Likewise immunotoxicology testing in one species may produce negative data in one species but positive data in another. Knowing the mechanistic basis through an understanding of species differences in the structure and function of the immune system is pivotal to success. This becomes especially true as pharmaceutical companies design and develop highly specific immunomodulatory molecules that demonstrate species-specific pharmacology. This review is an exploration of various species differences in the structure and function of the immune system and an attempt to identify those differences that may be important in the conduct of immunotoxicity tests. PMID- 12748042 TI - Determination of the immunotoxic potential of pesticides on functional activity of sheep leukocytes in vitro. AB - The effect of eight pesticides with different chemical structure (atrazine, bentazone, chloridazone, dichlofluanid, endosulfan, MCPA, simazine, triallate) on sheep peripheral blood phagocytes and lymphocytes was examined under in vitro conditions by iodo-nitro-tetrazolium reductase test and leukocyte migration inhibition assay. The pesticides, dissolved in DMSO, were tested at the concentrations of 10(-1)-10(-6) M. The significant suppression of metabolic activity of phagocytic cells was registered after exposure to dichlofluanid (10( 1)-10(-3) M), endosulfan, simazine and triallate (10(-1) M). The significant cytotoxic effect (the decrease of spontaneous migration of leukocytes) was registered for bentazone, dichlofluanid, endosulfan and MCPA (10(-1) M); chloridazone (10(-1) M-10(-2) M) and triallate (10(-1)-10(-5) M). The significant immunotoxic effect (the decrease of lymphocyte activation with PHA) was observed for atrazine (10(-1)-10(-2) M); bentazone (10(-2)-10(-4) M); dichlofluanid, endosulfan (10(-2)-10(-3) M); MCPA (10(-2)-10(-6) M) and simazine (10(-1)-10(-4) M). Three of the pesticides tested suppressed both, the metabolic activity of phagocytes and mitogenic activation of lymphocytes (dichlofluanid, endosulfan and simazine). Triallate suppressed the metabolic activity of phagocytes and showed a strong cytotoxic effect. Pesticides atrazine, bentazone and MCPA influenced the mitogenic activation of lymphocytes and chloridazone showed a significant cytotoxic effect. The different chemical structure of pesticides influenced the metabolic activity of phagocytic cells as well as mitogenic activation of lymphocytes to various intensity. PMID- 12748043 TI - Immunotoxicity of epicutaneously applied anticoagulant rodenticide warfarin: evaluation by contact hypersensitivity to DNCB in rats. AB - The immunotoxicity of epicutaneously administered anticoagulant rodenticide warfarin (WF) was examined in this work by using experimental contact hypersensitivity (CHS) reaction to hapten dinitrochlorobenzene (DNCB). WF (0.05 and 0.5 mg/kg) administration 24 h before the induction of CHS does not change expression of CHS evaluated by ear swelling assay. Regional draining lymph node response during sensitization phase was characterized by decreased cellularity but increased spontaneous and IL-2 stimulated proliferation of draining lymph node cells (DLC). No changes in IL-2 production and in numbers of CD25(+) cells were noted and even decreased proliferative index (ratio of IL-2 stimulated to unstimulated DLC proliferation) was detected. Increase in granulocyte activity (MTT reduction and adhesion to plastic) was noted following application of WF solely with further increase following subsequent application of DNCB, when granulocyte activation (NBT reduction) was noted also. Access of WF into general circulation might be responsible for observed changes, what was supported by ex vivo changes in DLC and granulocyte functions assessed before initiation of sensitization and by in vitro effect of exogenous WF as well. Differential effects of WF on lymphocytes and granulocytes noted in this study highlight the need for simultaneous testing of both cell type activity what might constitute a more integrated approach in immunotoxicity studies. PMID- 12748044 TI - Immunomodulation by cocaine and ketamine in postnatal rats. AB - The abuse of cocaine (COC) in combination with ketamine (KET) among pregnant women was shown to be high. Transplacental exposure is not the only route by which a newborn may be exposed to these agents, but they can also distribute into breast milk. Chronic COC exposure is associated with immunological modulation in human and animal models. The effect of sub-chronic exposure to COC and KET alone and in combination on the developing immune system was assessed in neonatal male Sprague-Dawley (SD) rats. To simulate the route of exposure during lactation, newborn male rats were treated orally with saline, COC alone (20 mg/kg), KET alone (50 mg/kg), or KET (50 mg/kg) followed 15 min later by COC (20 mg/kg) from days 1 to 21 of life. Pups were sacrificed 30 min following the last treatment. Total circulating leukocyte and lymphocyte counts were decreased with relative neutrophilia, while spleen/body weight ratio and IgM antibody response to sheep red blood cells (SRBCs) were increased in animals treated with COC. Moreover, treatment with COC alone increased serum interleukin 10 (IL-10) concentration; however, it did not affect serum interferon gamma (IFN-gamma) concentration. On the other hand, KET treatment did not produce any significant change of any of these parameters. However, when co-administered with COC, the immunomodulatory effects of COC were prevented. COC caused a significant increase in serum corticosterone concentration that KET effectively prevented. Lack of significant change of plasma and tissue concentrations of norcocaine (NC) suggested no role for COC metabolism in COC-induced immunomodulation. However, the results of this study indicate that COC-induced immunomodulatory reactions and their prevention by KET most likely occurred through neuroendocrinal mechanisms. PMID- 12748045 TI - The Kluyver effect revisited. AB - Yeast species can grow on various sugars. However, in many cases the growth on certain sugars (especially oligosaccharides) occurs only under aerobic conditions, and not in anaerobiosis or in the absence of respiration. Fermentation is blocked under these conditions. This apparent dependence of sugar utilization on the respiration has been called Kluyver effect, and such 'respiration-dependent' species are called Kluyver effect positive. A yeast may be Kluyver effect positive for some sugars and not for others. The physiological meaning and the molecular basis of the phenomenon are not clear. It has recently been reported that Kluyveromyces lactis, which is Kluyver effect positive for galactose and a few other sugars, could be converted into a Kluyver effect negative form by introduction of relevant sugar transporter genes. Such results offer for the first time a direct support to the hypothesis that the immediate cause of the Kluyver effect may be the low level of sugar transporter activities which is not sufficient to sustain the high substrate flow necessary for fermentative growth, whereas the energy-efficient respiratory growth does not require a high rate of sugar uptake. We examined to what extent this sugar transporter theory of the Kluyver effect can be generalized. PMID- 12748046 TI - Respiratory oscillations in yeast: mitochondrial reactive oxygen species, apoptosis and time; a hypothesis. AB - Oscillatory metabolic activities occur more widely than is generally realised; detectability requires observation over extended times of single yeast cells or synchrony of individuals to provide a coherent population. Where oscillations in intracellular metabolite concentrations are observed, the phenomenon has been ascribed to sloppy control, energetic optimisation, signalling, temporal compartmentation of incompatible reactions, or timekeeping functions. Here we emphasise the consequences of respiratory oscillations as a source of mitochondrially generated reactive O(2) metabolites. Temporal co-ordination of intracellular activities necessitates a time base. This is provided by an ultradian clock, and one result of its long-term operation is cyclic energisation of mitochondria, and thereby the generation of deleterious free radical species. Our hypothesis is that unrepaired cellular constituents and components (especially mitochondria) eventually lead to cellular senescence and apoptosis when a finite number of respiratory cycles has occurred. PMID- 12748047 TI - Role of nitric oxide in the response of Saccharomyces cerevisiae cells to heat shock and high hydrostatic pressure. AB - Nitric oxide (NO) is a simple and unique molecule that has diverse functions in organisms, including intracellular and intercellular messenger. The influence of NO on cell growth of Saccharomyces cerevisiae and as a signal molecule in stress response was evaluated. Respiring cells were more sensitive to an increase in intracellular NO concentration than fermentatively growing cells. Low levels of NO demonstrated a cytoprotective effect during stress from heat-shock or high hydrostatic pressure. Induction of NO synthase was isoform-specific and dependent on the metabolic state of the cells and the stress response pathway. These results support the hypothesis that an increase in intracellular NO concentration leads to stress protection. PMID- 12748048 TI - Outlines for the definition of halotolerance/halophily in yeasts: Candida versatilis (halophila) CBS4019 as the archetype? AB - Candida versatilis (halophila) CBS4019 was chosen to study the physiological reactions of long-term exposure to extremely high salt concentrations. In general, our results show a significant increase in enzyme expression during growth under stress conditions. Although glycerol and mannitol pathways are not under glucose repression, they were found to be metabolically regulated. Glycerol 3P-dehydrogenase used either of its cofactors NADPH or NADH, being in favor of NADPH during growth with high salt concentrations. This ability of interchanging cofactors, an increased fermentation rate, and the observed mannitol pathway activity are suggested to contribute to the yeasts' redox stability. Enzymes per se were not salt-tolerant in vitro. Consistently, intracellular sodium was low and intracellular potassium, a requirement for growth, was high. The concept of halophily and its applicability to yeasts is discussed. PMID- 12748049 TI - High-rate evolution of Saccharomyces sensu lato chromosomes. AB - Forty isolates belonging to the Saccharomyces sensu lato complex were analyzed for one nuclear and two mitochondrial sequences, and for their karyotypes. These data are useful for description and definition of yeast species based on the phylogenetic species concept. The deduced phylogenetic relationships among isolates based on the nuclear and mitochondrial sequences were usually similar, suggesting that horizontal transfer/introgression has not been frequent. The highest degree of polymorphism was observed at the chromosome level. Even isolates which had identical nuclear and mitochondrial sequences often exhibited variation in the number and size of their chromosomes. Apparently, yeast chromosomes have been frequently reshaped and therefore also the position of genes has been dynamic during the evolutionary history of yeasts. PMID- 12748050 TI - Genome-wide expression analyses: Metabolic adaptation of Saccharomyces cerevisiae to high sugar stress. AB - The transcriptional response of laboratory strains of Saccharomyces cerevisiae to salt or sorbitol stress has been well studied. These studies have yielded valuable data on how the yeast adapts to these stress conditions. However, S. cerevisiae is a saccharophilic fungus and in its natural environment this yeast encounters high concentrations of sugars. For the production of dessert wines, the sugar concentration may be as high as 50% (w/v). The metabolic pathways in S. cerevisiae under these fermentation conditions have not been studied and the transcriptional response of this yeast to sugar stress has not been investigated. High-density DNA microarrays showed that the transcription of 589 genes in an industrial strain of S. cerevisiae were affected more than two-fold in grape juice containing 40% (w/v) sugars (equimolar amounts of glucose and fructose). High sugar stress up-regulated the glycolytic and pentose phosphate pathway genes. The PDC6 gene, previously thought to encode a minor isozyme of pyruvate decarboxylase, was highly induced under these conditions. Gene expression profiles indicate that the oxidative and non-oxidative branches of the pentose phosphate pathway were up-regulated and might be used to shunt more glucose-6 phosphate and fructose-6-phosphate, respectively, from the glycolytic pathway into the pentose phosphate pathway. Structural genes involved in the formation of acetic acid from acetaldehyde, and succinic acid from glutamate, were also up regulated. Genes involved in de novo biosynthesis of purines, pyrimidines, histidine and lysine were down-regulated by sugar stress. PMID- 12748051 TI - ATP leakage from yeast cells treated by extracellular glycolipids of Pseudozyma fusiformata. AB - The ustilaginaceous yeast Pseudozyma fusiformata secreted glycolipids which were lethal to many yeasts and fungi more active at pH of about 4.0, and in the temperature range of 20-30 degrees C. Purified glycolipids enhanced non-specific permeability of the cytoplasmic membrane in sensitive cells, which resulted in ATP leakage and susceptibility of the cells to staining with bromocresol purple. Cells of Saccharomyces cerevisiae lost the ability to acidify the medium. Basidiomycetous yeasts were more sensitive to the glycolipids than ascomycetous ones. The minimal effective glycolipid concentration was 0.13 and 0.26 mg ml(-1) for Cryptococcus terreus and Filobasidiella neoformans, while for Candida albicans and Saccharomyces cerevisiae it was 1.0 and 1.6 mg ml(-1). PMID- 12748052 TI - Molecular typing of clinical and environmental Cryptococcus neoformans isolates in the Brazilian state Rio Grande do Sul. AB - In Brazil, 4.5% of the AIDS-related opportunistic infections are caused by Cryptococcus neoformans. This pathogen is a ubiquitous environmental basidiomycetous encapsulated yeast, commonly found in soil and avian excreta. The present study investigates further the population structure of clinical and environmental C. neoformans isolates from south Brazil. One hundred five clinical and 19 environmental (pigeon excreta and Eucalyptus spp.) isolates from the Brazilian state Rio Grande do Sul were characterized based on morphological, biochemical, molecular and serological data. The majority of the clinical and environmental isolates analyzed belonged to C. neoformans var. grubii serotype A (89.5 and 52.6%, respectively), were mating type alpha (98.1 and 94.7%, respectively) and were phospholipase-positive (94.3 and 73.7%, respectively). PCR fingerprinting with the microsatellite-specific primer M13 and the minisatellite specific primer (GACA)(4) grouped the majority of the isolates into the molecular type VNI (89.5 of the clinical and 52.6% of the environmental isolates). Our results add considerable new information to the few available data on ecology, molecular biology and epidemiology of C. neoformans in the southern region of Brazil. PMID- 12748053 TI - Phylogenetic relationships among yeasts of the 'Saccharomyces complex' determined from multigene sequence analyses. AB - Species of Saccharomyces, Arxiozyma, Eremothecium, Hanseniaspora (anamorph Kloeckera), Kazachstania, Kluyveromyces, Pachytichospora, Saccharomycodes, Tetrapisispora, Torulaspora, and Zygosaccharomyces, as well as three related anamorphic species assigned to Candida (C. castellii, C. glabrata, C. humilis), were phylogenetically analyzed from divergence in genes of the rDNA repeat (18S, 26S, ITS), single copy nuclear genes (translation elongation factor 1alpha, actin 1, RNA polymerase II) and mitochondrially encoded genes (small-subunit rDNA, cytochrome oxidase II). Single-gene phylogenies were congruent for well-supported terminal lineages but deeper branches were not well resolved. Analysis of combined gene sequences resolved the 75 species compared into 14 clades, many of which differ from currently circumscribed genera. PMID- 12748054 TI - Cloning and characterization of genes encoding trehalose-6-phosphate synthase (TPS1) and trehalose-6-phosphate phosphatase (TPS2) from Zygosaccharomyces rouxii. AB - In many organisms, trehalose protects against several environmental stresses, such as heat, desiccation, and salt, probably by stabilizing protein structures and lipid membranes. Trehalose synthesis in yeast is mediated by a complex of trehalose-6-phosphate synthase (TPS1) and trehalose-6-phosphate phosphatase (TPS2). In this study, genes encoding TPS1 and TPS2 were isolated from Zygosaccharomyces rouxii (designated ZrTPS1 and ZrTPS2, respectively). They were functionally identified by their complementation of the tps1 and tps2 yeast deletion mutants, which are unable to grow on glucose medium and with heat, respectively. Full-length ZrTPS1 cDNA is composed of 1476 nucleotides encoding a protein of 492 amino acids with a molecular mass of 56 kDa. ZrTPS2 cDNA consists of 2843 nucleotides with an open reading frame of 2700 bp, which encodes a polypeptide of 900 amino acids with a molecular mass of 104 kDa. The amino acid sequence encoded by ZrTPS1 has relatively high homology with TPS1 of Saccharomyces cerevisiae and Schizosaccharomyces pombe, compared with TPS2. Western blot analysis showed that the antibody against S. cerevisiae TPS1 recognizes ZrTPS1. Under normal growth conditions, ZrTPS1 and ZrTPS2 were highly and constitutively expressed, unlike S. cerevisiae TPS1 and TPS2. Salt stress and heat stress reduced the expression of the ZrTPS1 and ZrTPS2 genes, respectively. PMID- 12748055 TI - The origin of the cactus-yeast community. AB - The yeast community found in decaying cactus stems and cladodes is stable in terms of species membership and is similar in composition over space and time. The ecological origins of the three core and four common species in the assemblage were inferred by mapping yeast habitats onto a phylogeny of yeasts reconstructed from rDNA sequences. The members of the community belong to distinct clades and consequently have independent origins. The inferred evolutionary pathways of the taxa originate in either tree-flux or decaying fruit habitats and lead to decaying Opuntia cladode and columnar stem habitats. The reasons for the polyphyletic origins of the cactus-yeast community could be due to unique aspects of cactus chemistry, environmental extremes, vector association and interactions among the members. PMID- 12748056 TI - Myeloid related protein-8/14 stimulates interleukin-8 production in airway epithelial cells. AB - Excessive neutrophil recruitment is implicated in the pathogenesis of chronic lung diseases by causing collateral tissue damage. The cells move from the circulation in response to chemokines, such as interleukin (IL)-8, that are secreted by several lung cell types including epithelial cells. This study has investigated factors present in bronchial secretions that are responsible for IL 8 expression and secretion by epithelial cells and hence initiate or perpetuate the recruitment of neutrophils. A549 epithelial cells were stimulated with proinflammatory molecules likely to be of relevance in the lung. Tumor necrosis factor-alpha, IL-1beta, and lipopolysaccharide stimulated IL-8 production from epithelial cells in a dose- and time-dependent manner, and these effects were abrogated by specific antibodies or inhibitors. Bronchial secretions also stimulated IL-8 production, and lipopolysaccharide accounted for approximately 33% of this activity. An abundant 32-kD protein capable of stimulating IL-8 production was isolated from the secretion and identified as neutrophil cytoplasmic protein myeloid-related protein (MRP)-14, which is the heavy polypeptide chain in the MRP-8/14 heterodimer. Abrogation of MRP-14 activity with a specific antibody also reduced the IL-8-stimulating potential of bronchial secretions, suggesting it was a significant stimulus to IL-8 production in the lung and may amplify the neutrophilic inflammation seen in bronchial disease. PMID- 12748058 TI - Cigarette smoke prevents apoptosis through inhibition of caspase activation and induces necrosis. AB - Emphysema is characterized by enlargement of the distal airspaces in the lungs due to destruction of alveolar walls. Alveolar endothelial and epithelial cell apoptosis induced by cigarette smoke is thought to be a possible mechanism for this cell loss. In contrast, our studies show that cigarette smoke condensate (CSC) induces necrosis in alveolar epithelial cells and human umbilical vein endothelial cells. Furthermore, study of the cell death pathway in a model system using Jurkat cells revealed that in addition to inducing necrosis, CSC inhibited apoptosis induced by staurosporine or Fas ligation, with both effects prevented by the antioxidants glutathione and dithiothreitol. Time course experiments revealed that CSC inhibited an early step in the caspase cascade, whereby caspase 3 was not activated. Moreover, cell-free reconstitution of the apoptosome in cytoplasmic extracts from CSC-treated cells, by addition of cytochrome-c and dATP, did not result in activation of caspases-3 or -9. Thus, smoke treatment may alter the levels of pro- and antiapoptogenic factors downstream of the mitochondria to inhibit active apoptosome formation. Therefore, unlike previous studies, cell death in response to cigarette smoke by necrosis and not apoptosis may be responsible for the loss of alveolar walls and inflammation observed in emphysema. PMID- 12748057 TI - Apoptosis genes in human alveolar macrophages infected with virulent or attenuated Mycobacterium tuberculosis: a pivotal role for tumor necrosis factor. AB - Tumor necrosis factor (TNF)-alpha-dependent apoptosis of alveolar macrophages (AM) after infection with avirulent Mycobacterium tuberculosis (Mtb) results in bacillary death and the destruction of a growth niche for the pathogen. This response is minimized after infection with virulent strains of Mtb. To study the genetic control of Mtb-induced apoptosis, we used microarrays to interrogate the expression profile of infected human AM. Although we found variation in gene expression between different donors of AM, a set of genes were constant for each condition. A group of proapoptotic genes were downregulated after infection by virulent Mtb strain H37Rv, whereas infection with avirulent Mtb H37Ra led to a gene expression profile that would favor macrophage apoptosis. Neutralizing TNF in macrophage cultures infected with H37Ra changed the gene expression profile to one that resembled the profile of macrophages infected with H37Rv. These data reveal that apoptosis-related genes are regulated differently by virulent or attenuated Mtb strains, and are consistent with the hypothesis that virulent Mtb interfere with TNF death signaling. Given the importance of TNF in host defense against tuberculosis, the ability to repress the expression of genes activated by TNF may constitute a bacillary virulence mechanism. PMID- 12748059 TI - CCL20 is an inducible product of human airway epithelia with innate immune properties. AB - Chemokine ligand 20 (CCL20) and human beta-defensins (HBDs) share structural and functional properties, including antiparallel beta-pleated sheet core structures, charge distribution, and signaling to adaptive immune cells via the highly selective CCR6 receptor. Because of their similarities, we hypothesized that in addition to its known adaptive immune signaling functions, CCL20 has antimicrobial properties and participates in pulmonary innate immunity. We found that primary cultures of human airway epithelial and cultured fetal lung explants expressed CCL20 mRNA. Expression of CCL20 transcripts were significantly induced by interleukin (IL)-1beta and tumor necrosis factor-alpha, and inhibited by dexamethasone. Primary cultures of airway epithelia secreted CCL20 both apically and basolaterally, and CCL20 abundance was increased over 30-fold with IL-1beta stimulation, achieving an estimated concentration of 167 ng/ml in airway surface liquid. CCL20 abundance in bronchoalveolar lavage fluid from patients with cystic fibrosis was nearly 90-fold higher compared with bronchoalveolar lavage fluid from healthy volunteers. Interestingly, CCL20 exhibited salt-sensitive antimicrobial activity, mainly against Gram-negative bacteria in low mug/ml concentrations. Additionally, apical washings from IL-1beta-stimulated primary cultures of human airway epithelia had significantly more antimicrobial activity than unstimulated controls. CCL20 rapidly permeabilized bacterial membranes with a time course intermediate to HBD-2 and HBD-3. Thus, CCL20 is a bi-functional peptide with both innate and adaptive immune properties that is regulated by inflammatory mediators, expressed by airway epithelia, and increased in cystic fibrosis airway secretions. PMID- 12748060 TI - Cell-cell communication in heterocellular cultures of alveolar epithelial cells. AB - The mammalian alveolar epithelium is composed of alveolar type I (AT1) and alveolar type II (AT2) cells that together coordinate tissue function. We used a heterocellular culture model of AT1 and AT2 cells to determine pathways for intercellular signaling between these two phenotypes. Gap junction protein (connexin) profiles of AT1 and AT2 cells in heterocellular cultures were similar to those seen in rat lung alveolar sections. Dye coupling studies revealed functional gap junctions between and among each cell phenotype. Localized mechanical stimulation resulted in propagated changes of intracellular Ca2+ to AT1 or AT2 cells independent of the stimulated cell phenotype. Ca2+ communication that originated after AT1 cell stimulation was inhibited by gap junction blockers, but not by an inhibitor of extracellular nucleotide signaling (apyrase). Conversely, Ca2+ communication after stimulation of AT2 cells was not significantly reduced by gap junction inhibitors. However, apyrase significantly reduced Ca2+ communication from AT2 to AT1 cells, but not from AT2 to AT2 cells. In conclusion, AT1 and AT2 cells have unique connexin profiles that allow for functional coupling and distinct intercellular pathways for coordination of Ca2+ signaling. PMID- 12748061 TI - Feeding natural hydrophilic bile acids inhibits intestinal cholesterol absorption: studies in the gallstone-susceptible mouse. AB - We explored the influence of the hydrophilic-hydrophobic balance of a series of natural bile acids on cholesterol absorption in the mouse. Male C57L/J mice were fed standard chow or chow supplemented with 0.5% cholic; chenodeoxycholic; deoxycholic; dehydrocholic; hyocholic; hyodeoxycholic; alpha-, beta-, or omega muricholic; ursocholic; or ursodeoxycholic acids for 7 days. Biliary bile salts were measured by reverse-phase HPLC, and hydrophobicity indices were estimated by Heuman's method. Cholesterol absorption efficiency was determined by a plasma dual-isotope ratio method. In mice fed chow, natural proportions of tauro-beta muricholate (42 +/- 6%) and taurocholate (50 +/- 7%) with a hydrophobicity index of -0.35 +/- 0.04 produced cholesterol absorption of 37 +/- 5%. Because bacterial and especially hepatic biotransformations of specific bile acids occurred, hydrophobicity indices of the resultant bile salt pools differed from fed bile acids. We observed a significant positive correlation between hydrophobicity indices of the bile salt pool and percent cholesterol absorption. The principal mechanism whereby hydrophilic bile acids inhibit cholesterol absorption appears to be diminution of intraluminal micellar cholesterol solubilization. Gene expression of intestinal sterol efflux transporters Abcg5 and Abcg8 was upregulated by feeding cholic acid but not by hydrophilic beta-muricholic acid nor by hydrophobic deoxycholic acid. We conclude that the hydrophobicity of the bile salt pool predicts the effects of individual fed bile acids on intestinal cholesterol absorption. Natural alpha- and beta-muricholic acids are the most powerful inhibitors of cholesterol absorption in mice and might act as potent cholesterol-lowering agents for prevention of cholesterol deposition diseases in humans. PMID- 12748062 TI - PAR-2 modulates pepsinogen secretion from gastric-isolated chief cells. AB - In the present study, we investigated whether activation of protease-activated receptor type 2 (PAR-2) with SLIGRL (SL)NH2, a short mimetic agonistic peptide, directly stimulates pepsinogen secretion from gastric-isolated, pepsinogen secreting (chief) cells. Immunostaining of gastric-dispersed chief cells with a specific anti-PAR-2 antibody demonstrated expression of PAR-2 receptors on membrane and cytoplasm. SL-NH2 and trypsin potently stimulated pepsinogen secretion (EC50 = 0.3 nM) and caused Ca2+ mobilization (EC50 = 0.6 nM). In contrast to SL-NH2, the scramble peptide LSIGRL-NH2 failed to stimulate pepsinogen release. Exposure to SL-NH2 also resulted in ERK1/2 phosphorylation and activation. Exposure of chief cells to phosphotyrosine kinase inhibitors and 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one, a selective MEK inhibitor, significantly reduced secretion induced by SL-NH2. Pepsinogen secretion induced by SL-NH2 was desensitized by pretreating the cells with the mimetic peptide and trypsin, and exposure to SL-NH2 abrogates pepsinogen secretion induced by carbachol and CCK-8, but not secretion induced by secretin and vasointestinal peptide. Exposure to Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2 (substance P) but not to calcitonin gene-related peptide increased pepsinogen release. The neurokinin-1 receptor antagonist, N-acetyl-l-tryptophan 3,5 bis(trifluoromethyl)benzyl ester, inhibited substance P-stimulated pepsinogen secretion, whereas it did not affect secretion induced by SL-NH2. Collectively, these data indicate that PAR-2 is expressed on gastric chief cells and that its activation causes a Ca2+-ERK-dependent stimulation of pepsinogen secretion. PMID- 12748064 TI - A critical role for PKC zeta in endothelin-1-induced uterine contractions at the end of pregnancy. AB - We have previously shown that protein kinase C (PKC) zeta and/or PKC delta are necessary for endothelin-1 (ET-1)-induced human myometrial contraction at the end of pregnancy (Eude I, Paris P, Cabrol D, Ferre F, and Breuiller-Fouche M. Biol Reprod 63: 1567-1573, 2000). Here, we report that the selective inhibitor of PKC delta isoform, Rottlerin, does not prevent ET-1-induced contractions, whereas LY 294002, a phosphatidylinositol (PI) 3-kinase inhibitor, affects the contractile response. This study characterized the in vitro contractile response of cultured human pregnant myometrial cells to ET-1 known to induce in vitro contractions of intact uterine smooth muscle strips. Cultured myometrial cells incorporated into collagen lattices have the capacity to reduce the size of these lattices, referred to as lattice contraction. Neither the selective conventional PKC isoform inhibitor, Go-6976, or rottlerin affected myometrial cell-mediated gel contraction by ET-1, whereas this effect was blocked by LY-294002. We found that treatment of myometrial cell lattices with an inhibitory peptide specific for PKC zeta or with an antisense against PKC zeta resulted in a significant loss of ET-1 induced contraction. Evidence is also presented by using confocal microscopy that ET-1 induced translocation of PKC zeta to a structure coincident with the actin rich microfilaments of the cytoskeleton. We have shown that PKC zeta has a role in the actin organization in ET-1-stimulated cells. Accordingly, our results suggest that PKC zeta plays a role in myometrial contraction in pregnant women. PMID- 12748063 TI - TNF-alpha-induced cell death in ethanol-exposed cells depends on p38 MAPK signaling but is independent of Bid and caspase-8. AB - Alcoholic liver disease is associated with an increase in the number of necrotic and apoptotic liver parenchymal cells. Part of this injury is mediated by TNF alpha. Ethanol exposure sensitizes cells to the cytotoxic effects of TNF-alpha. This may be due, in part, to the increased propensity of the mitochondria in ethanol-exposed cells to induction of mitochondrial permeability transition (MPT) by various agents, including the proapoptotic protein Bax. This idea is supported by the observation that increased cell death induced by TNF-alpha in ethanol exposed cells was dependent on development of the MPT. In the present study, we elucidate the pathways through which ethanol exposure enhances TNF-alpha induction of the MPT and the resulting cytotoxicity. Specifically, ethanol exposed cells display caspase-8- and Bid-independent cell killing during TNF alpha treatment. Moreover, the ethanol-enhanced pathway is dependent on p38 MAPK signaling, which brings about caspase-3 activation, mitochondrial depolarization, accumulation of cytochrome c in the cytosol, and the translocation of Bax to the mitochondria. Additionally, ethanol-exposed cells display a blunting of TNF-alpha induced Akt activation and Bcl-2 antagonist of cell death phosphorylation that may account, in part, for the increased sensitivity of the mitochondria to Bax mediated damage. PMID- 12748065 TI - Hyperosmotic stress activates Rho: differential involvement in Rho kinase dependent MLC phosphorylation and NKCC activation. AB - Hyperosmotic stress initiates adaptive responses, including phosphorylation of myosin light chain (MLC) and concomitant activation of Na+-K+-Cl- cotransporter (NKCC). Because the small GTPase Rho is a key regulator of MLC phosphorylation, we investigated 1) whether Rho is activated by hyperosmotic stress, and if so, what the triggering factors are, and 2) whether the Rho/Rho kinase (ROK) pathway is involved in MLC phosphorylation and NKCC activation. Rho activity was measured in tubular epithelial cells by affinity pulldown assay. Hyperosmolarity induced rapid (<1 min) and sustained (>20 min) Rho activation that was proportional to the osmotic concentration and reversed within minutes upon restoration of isotonicity. Both decreased cell volume at constant ionic strength and elevated total ionic strength at constant cell volume were capable of activating Rho. Changes in [Na+] and [K+] at normal total salinity failed to activate Rho, and Cl depletion did not affect the hyperosmotic response. Thus alterations in cellular volume and ionic strength but not individual ion concentrations seem to be the critical triggering factors. Hyperosmolarity induced mono- and diphosphorylation of MLC, which was abrogated by the Rho-family blocker Clostridium toxin B. ROK inhibitor Y-27632 suppressed MLC phosphorylation under isotonic conditions and prevented its rise over isotonic levels in hypertonically stimulated cells. ML-7 had a smaller inhibitory effect. In contrast, it abolished the hypertonic activation of NKCC, whereas Y-27632 failed to inhibit this response. Thus hyperosmolarity activates Rho, and Rho/ROK pathway contributes to basal and hyperosmotic MLC phosphorylation. However, the hypertonic activation of NKCC is ROK independent, implying that the ROK-dependent component of MLC phosphorylation can be uncoupled from NKCC activation. PMID- 12748066 TI - Nicotinic acetylcholine receptor alpha 7 regulates cAMP signal within lipid rafts. AB - Neuronal nicotinic acetylcholine receptors (nAChRs) are made of multiple subunits with diversified functions. The nAChR alpha 7-subunit has a property of high Ca2+ permeability and may have specific functions and localization within the plasma membrane as a signal transduction molecule. In PC-12 cells, fractionation by sucrose gradient centrifugation revealed that nAChR alpha 7 existed in low density, cholesterol-enriched plasma membrane microdomains known as lipid rafts where flotillin also exists. In contrast, nAChR alpha 5- and beta2-subunits were located in high-density fractions, out of the lipid rafts. Type 6 adenylyl cyclase (AC6), a calcium-inhibitable isoform, was also found in lipid rafts and was coimmunoprecipitated with nAChR alpha 7. Cholesterol depletion from plasma membranes with methyl-beta-cyclodextrin redistributed nAChR alpha 7 and AC6 diffusely within plasma membranes. Nicotine stimulation reduced forskolin stimulated AC activity by 35%, and this inhibition was negated by either treatment with alpha-bungarotoxin, a specific antagonist of nAChR alpha 7, or cholesterol depletion from plasma membranes. The effect of cholesterol depletion was negated by the addition of cholesterol. These data suggest that nAChR alpha 7 has a specific membrane localization relative to other nAChR subunits and that lipid rafts are necessary to localize nAChR alpha 7 with AC within plasma membranes. In addition, nAChR alpha 7 may regulate the AC activity via Ca2+ within lipid rafts. PMID- 12748067 TI - Cloning of a Na+-driven Cl/HCO3 exchanger from squid giant fiber lobe. AB - We extracted RNA from the giant fiber lobe (GFL) of the squid Loligo pealei and performed PCR with degenerate primers that were based on highly conserved regions of Na+-coupled HCO3- transporters. This approach yielded a novel, 290-bp sequence related to the bicarbonate transporter superfamily. Using an L. opalescens library, we extended the initial fragment in the 3' and 5' directions by a combination of library screening and PCR and obtained the full-length clone (1,198 amino acids) by PCR from L. pealei GFL. The amino acid sequence is 46% identical to mammalian electrogenic and electroneutral Na-HCO3 cotransporters and 33% identical to the anion exchanger AE1. Northern blot analysis showed strong signals in L. pealei GFL, optic lobe, and heart and weaker signals in gill and stellate ganglion. To assess function, we injected in vitro-transcribed cRNA into Xenopus oocytes and subsequently used microelectrodes to monitor intracellular pH (pHi) and membrane voltage (Vm). Superfusing these oocytes with 5% CO2-33 mM HCO3 caused a CO2-induced fall in pHi, followed by a slow recovery. The absence of a rapid HCO3- -induced hyperpolarization indicates that the pHi recovery mechanism is electroneutral. Ion substitutions showed that Na+ and Cl- are required on opposite sides of the membrane. Transport was blocked by 50 microM 4,4' diisothiocyanostilbene-2,2'-disulfonic acid (DIDS). The characteristics of our novel clone fit those of a Na+-driven Cl/HCO3 exchanger (NDCBE). PMID- 12748068 TI - Increased phosphorylation of myosin light chain associated with slow-to-fast transition in rat soleus. AB - In striated muscles myosin light chain (MLC)2 phosphorylation regulates calcium sensitivity and mediates sarcomere organization. Little is known about the changes in MLC2 phosphorylation in relation to skeletal muscle plasticity. We studied changes in MLC2 phosphorylation in rats receiving three treatment conditions causing slow-to-fast transitions: 1) atrophy induced by 14 days of hindlimb suspension (HS), 2) hypertrophy induced by 14 days of clenbuterol administration (CB), and 3) 14 days of combined treatment (CB-HS). Three variants of the slow (MLC2s) and two variants of the fast MLC2 (MLC2f) isoform were separated with two-dimensional electrophoresis and identified with monoclonal and polyclonal antibodies specific for MLC2; their relative proportions were densitometrically quantified. In control soleus muscle MLC2s predominated over MLC2f (91.4 +/- 3.9% vs. 8.5 +/- 3.9%) and was separated into two spots, the less acidic spot being 73.5 +/- 4.3% of the total. All treatments caused a decrease of the less acidic unphosphorylated spot of MLC2s (CB: 64.1 +/- 5.6%, HS: 62.4 +/- 6.8%, CB-HS: 56.4 +/- 4.4%), the appearance of a third more acidic variant of MLC2s (representing 3.9-5.9% of total MLC2s), an increase of MLC2f (CB: 30.9 +/- 3.1%, HS: 23.9 +/- 3.3%, CB-HS: 25.3 +/- 3.9%), and the phosphorylation of a large fraction of MLC2f (CB: 30.4 +/- 6.7%, HS: 28.7 +/- 6.5%, CB-HS: 21.8 +/- 2.1%). Treatment with alkaline phosphatase or with protein phosphatase 1 (PP1) removed the most acidic spots of both MLC2f and MLC2s. We conclude that in rat skeletal muscles an increase of MLC2 phosphorylation is associated with the slow to-fast transition regardless of whether hypertrophy or atrophy develops. PMID- 12748069 TI - Higher field strength for proton MR spectroscopy. PMID- 12748070 TI - Glial neoplasms without elevated choline-creatine ratios. PMID- 12748071 TI - Measuring the effect of novel therapies for back pain. PMID- 12748072 TI - The changing MR imaging appearance of polymicrogyria: a consequence of myelination. AB - BACKGROUND AND PURPOSE: During the review of MR studies of multiple patients with polymicrogyria (PMG), it was noted that the patterns of cortical abnormality differed significantly among affected patients. In particular, the cortex appeared very thin in some patients, but was thick in others. The purpose of the present study was to attempt to clarify the cause of the different imaging appearances. METHODS: T1- and T2-weighted images obtained in 17 patients (age range, 3 days to 43 years) with PMG diagnosed on the basis of imaging characteristics were retrospectively reviewed. One patient was examined four times over a period of 21 months. Particular attention was paid to the thickness and signal intensity of the cortex and underlying white matter and how these features varied with maturation of the cortex and white matter. RESULTS: T2 weighted images revealed two patterns of PMG. Pattern 1 showed small, fine, and undulating cortex with normal thickness (3-4 mm) in seven patients, all younger than 12 months; and pattern 2, a bumpy cortex that appeared abnormally thick (6-8 mm) and had an irregular cortical-white matter junction in seven patients older than 18 months. Both patterns were observed in four patients between 15 months and 2 years of age (ie, pattern 1 in the anterior frontal region and pattern 2 in the posterior frontal, parietal, or perisylvian regions). A layer of T2 prolongation (2-3 mm) was recognized between pattern 1 PMG and underlying myelinated white matter in four patients 11 months to 2 years of age. T1-weighted images showed either poor differentiation of the cortex and underlying white matter or pattern 2. Serial MR imaging in one patient depicted longitudinal changes of the PMG from pattern 1 to pattern 2. CONCLUSION: These findings suggest that the two appearances (thin and thick) of the cortex seen in PMG likely represent the same process, with the apparent difference being the result of myelination in subcortical and intracortical fibers that cause a change of the appearance and apparent thickness of PMG on T2-weighted images. PMID- 12748073 TI - Brain imaging in the unsedated pediatric patient: comparison of periodically rotated overlapping parallel lines with enhanced reconstruction and single-shot fast spin-echo sequences. AB - BACKGROUND AND PURPOSE: Periodically rotated overlapping parallel lines with enhanced reconstruction (PROPELLER or PROP) is an effective means of compensating for head motion during MR imaging in adults. The aim of this study was to assess the value of this novel technique in unsedated children. METHODS: PROP T2 weighted fast spin-echo (FSE) imaging (TR/TE/NEX, 4000/83/2; 50 seconds) and T2 weighted single-shot FSE (SS-FSE) imaging (19,929/92/0.5; imaging time, 25 seconds) were performed in 35 unsedated children (mean age, 4.7 years +/- 4.2) who were undergoing brain MR imaging. Two observers assessed unlabelled images for motion artifact, other artifacts, visibility of pathology, and the preferred image overall. Sequences were compared by using the chi(2) test and concordant data from both observers. RESULTS: Both PROP and the SS-FSE imaging offered equal degrees of motion correction. Metallic artifacts were worse on PROP imaging, likely because of a higher receiver bandwidth (P <.001, chi(2) test). Pathology was present in 28 subjects and equally well seen on PROP and SS-FSE images. Overall, PROP was preferred, largely because of its improvements in image contrast (P <.001, chi(2) test). CONCLUSION: SS-FSE imaging and PROP provide equal motion correction, although PROP enables better assessment of the brain parenchyma. PMID- 12748074 TI - Apparent diffusion coefficient determination in normal fetal brain: a prenatal MR imaging study. AB - BACKGROUND AND PURPOSE: Diffusion-weighted MR imaging studies of normal brain development have focused on premature babies who were free of focal lesions on conventional MR images. The condition of prematurity, however, is dissimilar to intrauterine life. We sought to establish normal values of fetal brain apparent diffusion coefficient (ADC) to highlight its abnormal changes in pathologic conditions and to obtain information about normal brain development. METHODS: We measured the ADC, in utero, by using an echo-planar three-axes diffusion sensitized sequence (b factor, 0 and 600 s/mm(2)), in frontal and occipital white matter and basal ganglia gray matter of 15 fetuses. Their gestational ages ranged from 22 to 35 weeks, and the postnatal MR images or sonograms revealed normal brain. RESULTS: Mean ADC value was 1.96 +/- 0.1 micro m(2)/ms (SD) in frontal white matter, 1.95 +/- 0.1 micro m(2)/ms in occipital white matter, and 1.56 +/- 0.1 micro m(2)/ms in basal ganglia. A significant negative correlation between ADC and gestational age was found for basal ganglia, whereas only a trend was present for frontal white matter. CONCLUSION: Although moderately higher, the ADC determinations we obtained are consistent with those reported in the literature in postnatal studies performed in premature babies. PMID- 12748075 TI - White matter injury in the premature infant: a comparison between serial cranial sonographic and MR findings at term. AB - BACKGROUND AND PURPOSE: The accuracy of cranial sonography (US) in characterizing white matter (WM) injury in the premature infant is unclear. This study was aimed to assess the sensitivity and specificity of serial cranial US during the first 6 weeks of life in comparison to MR imaging at term (week of expected delivery) in characterizing the presence of WM injury in a cohort of 96 very low birth weight (VLBW) infants. METHOD: A blinded investigator reviewed serial cranial sonograms for the presence of WM echolucency and echodensity, including its duration and extent. These abnormalities were compared with a second independent investigator's evaluation to determine the sensitivity and specificity of cranial WM abnormalities at US. RESULTS: The presence of prolonged echodensity (>7 days) in the WM on neonatal cranial sonograms demonstrated low sensitivity (26%) and a low positive predictive value (36%) for the presence of noncystic WM injury, as detected on MR images at term. Extensive cystic lesions detected on MR images were all identified during earlier cranial US. CONCLUSION: Neonatal cranial US of the VLBW infant demonstrates high reliability in the detection of cystic WM injury but has significant limitations in the demonstration of noncystic WM injury. This deficiency of neonatal cranial US is important, because noncystic WM injury is considerably more common than cystic WM injury. PMID- 12748076 TI - Neoangiogenesis in association with moyamoya syndrome shown by estimation of relative recirculation based on dynamic contrast-enhanced MR images. AB - BACKGROUND AND PURPOSE: This study examined abnormalities of the recirculation phase of the contrast medium concentration time course curve in patients with moyamoya disease by using the relative recirculation (rR) parameter. Our aims were to increase understanding of the rR parameter and to assess its possible value in the investigation of moyamoya disease. METHODS: Six patients with confirmed moyamoya disease and five normal participants were examined by using dynamic T2*-weighted MR perfusion imaging. Parametric maps of relative cerebral blood volume, time to peak (TTP), and rR were calculated. The width of the normal distribution of TTP and the normal upper limit for rR were derived from the normal participants by identification of the 97.5% confidence limits for the distributions. Major vessels were identified by selection of pixels with relative cerebral blood volume >0.5. Areas of abnormally prolonged TTP and of increased rR were then identified in patients, and the relationship between these parameters and major vessels was examined. RESULTS: The combination of cerebral blood volume >50%, elevated TTP, and elevated rR values accurately identified areas of moyamoya vessel formation seen on angiograms. Larger well-developed meningeal and basal collaterals did not show abnormalities of rR. Areas of prolonged TTP were also observed to occur in the brain distal to arterial occlusion in all cases. In three of these, abnormal elevation of rR was observed in the center of regions of prolonged TTP. CONCLUSION: These findings support those of our previous work, which suggested that rR can act as an indicator of regional microvascular abnormality, such as that seen in areas of angiogenic activity. They also suggest that measurement of rR may provide valuable additional information regarding patients with moyamoya disease. PMID- 12748077 TI - Diffusion tensor imaging in cases of adrenoleukodystrophy: preliminary experience as a marker for early demyelination? AB - BACKGROUND AND PURPOSE: Diffusion tensor imaging measures the proton diffusivity and preferential orientation of the diffusion tensor. X-linked adrenoleukodystrophy is a demyelinating disease for which therapy depends on the onset and extension of demyelination. We investigated the ability of diffusion tensor imaging to detect changes in the demyelinated lesions and in the normal appearing white matter. METHODS: Diffusion tensor imaging of three related boys with X-linked adrenoleukodystrophy and seven age-matched control participants was performed. Isotropic diffusion (D') and fractional anisotropy (FA) values were determined in 18 regions of interest in the white matter of both hemispheres. RESULTS: In all the demyelinated white matter areas, a pattern with increased D' and loss of FA was found. For example, mean D' was 1.772 x 10(-3)mm(2)/s in patient 2 with blindness and extensive demyelination of the occipital white matter and was 0.693 x 10(-3)mm(2)/s in control participants (P =.01). In the same region, mean FA was 0.103 (0.464 in control participants, P <.0001). Significant alterations of D' and FA were also observed in normal appearing white matter. For example, mean D' was 0.802 x 10(-3)mm(2)/s in the parietal white matter of patient 1 with no visible alterations on T2-weighted images (0.715 x 10(-3) mm(2)/s in control patients, P =.03), whereas mean FA was 0.320 (0.400 in control participants, P =.003). CONCLUSION: Elevated D' and loss of FA revealed by diffusion tensor imaging are consistent with severe demyelination in patients with X-linked adrenoleukodystrophy. Significant alterations of D' and FA in normal appearing white matter may indicate early demyelination in areas that are not yet visibly altered on conventional MR images. Further evaluation in a larger series of patients and long-term study are needed. PMID- 12748079 TI - The usefulness of MR imaging in the diagnosis of dysembryoplastic neuroepithelial tumor in children: a study of 14 cases. AB - BACKGROUND AND PURPOSE: Dysembryoplastic neuroepithelial tumors (DNTs) are benign lesions affecting children and are associated with epilepsy. The goal of our study was to better characterize the clinical-radiologic-pathologic spectrum of DNTs (complex and simple forms only) in a series of 14 children. METHODS: Clinical, neuroradiologic, and pathologic features of all cases were retrospectively studied. RESULTS: Eleven cases of complex and three cases of simple DNTs were identified. Mean follow-up was 87 months, and no recurrence was recorded except for one case of simple DNT. We found that some neuroradiologic features may be helpful to support the diagnosis of DNT: presence of "septations," triangular pattern of distribution, and absence of contrast enhancement. CONCLUSION: The evidence of the specific glioneuronal element is found by pathologic examination, but the typical neuroradiologic aspect of DNT suggests this diagnosis preoperatively. Radiologic examination may be helpful for the diagnosis of DNT when pathologic findings are inconclusive. PMID- 12748078 TI - MR imaging features in Marinesco-Sjogren syndrome: severe cerebellar atrophy is not an obligatory finding. AB - BACKGROUND AND PURPOSE: Cerebellar atrophy is considered the most prominent neuroradiologic finding in Marinesco-Sjogren syndrome (MSS). Our purpose was to investigate this neuroradiologic feature in a series of patients with MSS. METHODS: Five patients with MSS (age range, 5-19 years) underwent native MR imaging of the brain. The findings were assessed with particular attention to the cerebellum and the supratentorial structures. RESULTS: Only two patients had slight cerebellar atrophy; the cerebellum was normal in size and configuration in the other patients. Additional supratentorial findings were present in some of the patients, with an apparently small anterior pituitary gland in two and the absence of the posterior pituitary bright spot in three of the patients. CONCLUSION: Cerebellar atrophy is not an obligatory finding in MSS, and almost normal cranial MR imaging results are compatible with the diagnosis. Morphologic changes of the pituitary gland seem to be common in patients with MSS and are not associated with endocrine dysfunction. PMID- 12748080 TI - Tuberous sclerosis and fibrous dysplasia. AB - We present the case of an 11-year-old female patient with tuberous sclerosis who had a right nasal mass. CT examination revealed fibrous dysplasia involving the frontal, ethmoid, sphenoid, and vomer bones. Biopsy findings of the mass confirmed this diagnosis, and follow-up revealed marked expansion of these lesions. The authors emphasize the association of bone abnormalities and tuberous sclerosis and discuss the consideration of fibrous dysplasia as a component of this syndrome. PMID- 12748081 TI - Meningocele-induced positional syncope and retinal hemorrhage. AB - Meningocele is recognized as a rare, usually asymptomatic condition not associated with acute neurologic symptoms. We herein describe the case of a patient with a longstanding history of a lower back "mass" and recurrent syncope who became acutely unresponsive and developed bilateral retinal hemorrhages when she was placed in the supine position to undergo carotid sonography. MR imaging revealed a large, dorsal lumbar meningocele. The episode likely was caused by acutely increased intracranial pressure caused by displacement of CSF from the meningocele intracranially. PMID- 12748082 TI - Unusual presentation of GM2 gangliosidosis mimicking a brain stem tumor in a 3 year-old girl. AB - We report a case of GM2 gangliosidosis revealed by MR imaging of an isolated brain stem abnormality in a 3-year-old girl referred for gait difficulties related to ataxia and pyramidal signs. Brain MR imaging displayed a brain stem lesion with high signal intensity on fluid-attenuated inversion recovery and T2 weighted images, suggesting either a tumor or an inflammatory process. Stereotactic biopsy findings showed the presence of swollen neurons with storage material in lysosomes. Enzyme study revealed deficiency of hexosaminidase A, variant B1. Gangliosidoses should be considered in the differential diagnosis of isolated infiltrating brain stem lesions in childhood. PMID- 12748083 TI - Proton MR spectroscopy in mild cognitive impairment and Alzheimer disease: comparison of 1.5 and 3 T. AB - BACKGROUND AND PURPOSE: Theoretically, proton ((1)H) MR spectroscopy at a higher field strength has the advantages of higher signal-to-noise ratio and improved spectral resolution. We therefore compared the ability of single-voxel (1)H MR spectroscopy at 1.5 and 3 T to diagnostically discriminate among cognitively normal elderly subjects, patients with mild cognitive impairment (MCI), and patients with Alzheimer disease (AD). METHODS: At both 1.5 and 3 T, we studied 41 cognitively normal elderly subjects, 20 patients with MCI, and 20 patients with AD. In each subject, (1)H MR spectroscopy was performed at TEs of 30 and 135 ms and from voxels placed over the posterior cingulate gyri. RESULTS: Average line widths and interexamination variability of metabolite ratios were higher at 3 T than at 1.5 T. Consistent quantification of glutamine (Gln) + glutamate/creatine (Cr) and Gln/Cr peak ratios occurred at 3 T but not at 1.5 T. Choline (Cho)/Cr (at TE = 135 ms) and myo-inositol (MI)/Cr were higher and N-acetylaspartate (NAA)/Cr (at TE = 135 ms) and NAA/MI were lower in patients with MCI than in cognitively normal subjects only at 1.5 T. MI/Cr and Cho/Cr were higher and NAA/Cr and NAA/MI were lower in patients with AD than in cognitively normal subjects at both 1.5 and 3 T. Differentiation of patients with AD from cognitively normal subjects by using the NAA/MI data was similar at both field strengths (P >.05). CONCLUSION: With currently available technology, the diagnostic performance of (1)H MR spectroscopy in patients with MCI and those with AD was not better at 3 T than at 1.5 T. PMID- 12748084 TI - Abnormal brain diffusivity in patients with neuropsychiatric systemic lupus erythematosus. AB - BACKGROUND AND PURPOSE: Neuroimaging techniques have increased our knowledge of the pathogenesis of neuropsychiatric systemic lupus erythematosus (NPSLE) and have been useful in supporting the diagnosis. Nevertheless, new imaging techniques are needed to unravel the exact pathogenesis and to provide diagnostic criteria for NPSLE. In this preliminary study, we investigated whether diffusion weighted imaging (DWI) can depict cerebral abnormalities in patients with a history of NPSLE, and we assessed whether apparent diffusion coefficient (ADC) histograms in these patients differ from those of healthy control subjects. METHODS: Eleven female patients with a history of NPSLE (mean age [+/- SD], 35 years +/- 9) and 10 healthy control subjects (eight female, two male; mean age, 37 years +/- 16) underwent DWI. DWI and ADC images were assessed by means of visual inspection, and histograms were composed from the ADC images. From these, we derived a variety of parameters that quantitatively reflect the diffusivity of brain parenchyma. RESULTS: Visual inspection of ADC images and DWIs did not reveal any abnormalities in either patients with NPSLE or control subjects. In contrast, ADC histograms of the NPSLE group were, on average, significantly lower and broader, with a higher mean ADC value. CONCLUSION: The data suggest an increased general diffusivity in brain parenchyma of patients with NPSLE, probably based on loss of tissue integrity. In addition to increasing our battery of highly wanted diagnostic tools and our understanding of the pathogenesis of NPSLE, the present method seems to be useful in quantifying the disease burden, enabling monitoring in treatment trials and the study of disease progression. PMID- 12748085 TI - Focal lesion in the splenium of the corpus callosum on FLAIR MR images: a common finding with aging and after brain radiation therapy. AB - BACKGROUND AND PURPOSE: Focal high signal intensity in the splenium of the corpus callosum on fluid-attenuated inversion-recovery (FLAIR) images is generally considered an abnormal MR finding. We identified high signal intensity in the splenium on FLAIR images in patients of advanced age with otherwise normal images and in patients who had received brain radiation therapy. We undertook an investigation to determine the frequency of this finding in these patient groups. METHODS: We reviewed the FLAIR images and medical records of 67 patients (group 1) imaged for suspicion of CNS disease and of 18 consecutive patients (group 2) with history of brain radiation therapy. All FLAIR images were evaluated for focal signal intensity abnormalities in the splenium and for diffuse white matter abnormalities. Also, autopsy specimens from two cases not part of either study group were examined. RESULTS: Among the initial 67 patients in group 1, focal high signal intensity in the splenium was associated with aging, radiation therapy, and white matter changes. Focal high signal intensity in the splenium was evident on FLAIR images in 16 of the 18 patients in the post-radiation therapy group. Histologic examination of the splenium in one autopsy case with a history of chest and neck radiation therapy demonstrated isomorphic gliosis. CONCLUSION: High signal intensity in the splenium of the corpus callosum on FLAIR images is a common finding after brain radiation therapy and can be seen with aging. The radiologist should be aware of this common finding and not mistake it for more commonly recognized causes of splenial lesions. PMID- 12748086 TI - Measurement of cerebral blood flow in chronic carotid occlusive disease: comparison of dynamic susceptibility contrast perfusion MR imaging with positron emission tomography. AB - BACKGROUND AND PURPOSE: Our purpose was to evaluate the accuracy of cerebral blood flow (CBF) measurements obtained by using dynamic susceptibility contrast enhanced MR imaging, including the influence of arterial input function (AIF) selection, compared with those obtained by using [(15)O]-H(2)O positron emission tomography (PET) for patients with chronic carotid occlusion. METHODS: MR images and PET scans were obtained of seven patients with unilateral carotid occlusion and were co-registered for region of interest analysis. PET CBF maps were generated by using the autoradiographic method. MR imaging CBF maps were calculated by deconvolution of the susceptibility time curve with a proximal middle cerebral artery AIF and were converted to absolute flow rates either by assuming a constant contralateral white matter CBF value of 22 mL/100 mL/min or by using individually determined PET white matter CBF values. RESULTS: Although CBF values measured by PET and MR imaging were positively correlated for every patient, the slopes and y intercepts of the regression lines varied widely among patients. The correlation was better when individual white matter CBF values measured by PET were used to scale the white matter CBF values measured by MR imaging (r = 0.84, P <.0001) than when constant contralateral CBF values were assumed (r = 0.54, P <.0001). The choice of AIF ipsilateral or contralateral to the occluded carotid artery made no statistically significant difference (P >.05) to the correlation coefficient, slope, or y intercept of the MR imaging versus PET CBF regressions for six of the seven patients. CONCLUSION: Although linearly correlated with CBF values measured by PET, dynamic susceptibility contrast enhanced MR imaging was not accurate for measuring absolute CBF values. AIF selection relative to the side of carotid occlusion did not significantly affect calculated MR imaging CBF values for six of the seven patients. PMID- 12748087 TI - Cavernous carotid artery calcification and white matter ischemia. AB - BACKGROUND AND PURPOSE: The relevance of cavernous carotid artery calcification on unenhanced CT scans of the brain has recently been investigated against the backdrop of the widespread implementation of coronary artery calcification scoring. We sought to determine whether the degree of cavernous carotid artery calcification correlated with scores of white matter hyperintensity seen on MR images. In so doing, we sought to establish a relative risk for future stroke on the grade of carotid calcification. METHODS: Neuroradiologic findings in 187 patients who underwent CT and MR imaging examinations within 1 month of each other were retrospectively reviewed. The degree of circumferential calcification and thickness of calcification were graded for the cavernous carotid arteries on the basis of CT findings. Using the scale developed by the Cardiovascular Health Study, the white matter was graded for degree of disease on the basis of MR findings. Correlation tests and regression analyses were performed to determine the impact of age, race, and sex on results. RESULTS: Although the cavernous carotid calcification scores and the MR imaging white matter scores showed good correlation (P <.001), the effect was mediated by age. With age factored in as a covariant, no correlation was shown between CT calcification scores and MR imaging white matter scores. Sex had no effect, but African American study participants had worse MR imaging white matter scores than did white participants. CONCLUSION: After adjusting for age, cavernous carotid calcification grades and MR imaging white matter scores do not show a significant correlation. The relative risk for future stroke cannot be predicted from cavernous carotid calcifications. PMID- 12748088 TI - Sensitivity and interrater agreement of CT and diffusion-weighted MR imaging in hyperacute stroke. AB - BACKGROUND AND PURPOSE: Previous acute stroke studies found diffusion-weighted (DW) imaging superior to CT for detection of early ischemic signs (EIS). However, these findings were confounded by a large time interval in favor of DW imaging. We compared DW images and CT scans obtained with a short time delay in patients with acute stroke to define the sensitivity and interrater agreement of both imaging techniques. METHODS: CT scans and DW images were obtained within 6 hours of symptom onset in 46 patients with acute stroke. Three neuroradiologists and three neurologists reviewed the images for EIS in five regions of the middle cerebral artery (MCA) territory and estimated the extent of EIS (< or > one-third of the MCA territory). RESULTS: The mean delay between imaging with both modalities was 24.5 minutes (range, 10-41 minutes). Forty-five of 46 patients had an ischemic stroke. EIS were seen on 33 of 45 CT scans (73% sensitivity; 95% confidence interval [CI]: 58-85%) and on 42 of 45 DW images (93% sensitivity; 94% CI: 82-99%). Interrater agreement was moderate (kappa = 0.57) for CT and excellent (kappa = 0.85) for DW imaging. CT studies had a moderate interrater agreement for estimation of EIS greater than one-third of the MCA territory (kappa = 0.40), whereas DW imaging showed good results (kappa = 0.68). Sensitivity for detection of greater than one-third of the MCA territory was equally poor (57%, 95% CI: 29-82%) for both CT and DW imaging. CONCLUSION: DW imaging helped identify EIS with higher sensitivity than that of CT. The interrater variability of the one-third rule was high for CT, and thus the clinical applicability of CT is limited. Our results support the application of stroke MR imaging for the treatment of patients with acute stroke. PMID- 12748089 TI - Ultrasonographically predicting the extent of collateral flow through superficial temporal artery-to-middle cerebral artery anastomosis. AB - BACKGROUND AND PURPOSE: This study was performed to elucidate whether the extent of bypass flow through superficial temporal artery-to-middle cerebral artery (STA MCA) anastomosis could be indirectly estimated by measuring the blood flow velocity in the superficial temporal artery (STA) by using duplex ultrasonography. METHODS: We analyzed 29 patients (31 sides) who underwent STA MCA bypass surgery for occlusive cerebrovascular disease (28 sides) or unclippable cerebral aneurysm that required therapeutic occlusion of the internal carotid artery (three sides). The flow velocities of the STA were measured by using ultrasonography. For patients who underwent the surgery unilaterally, the flow velocity ratios of the operated side to the contralateral side for the individual arteries were calculated. The correlation between these flow velocity parameters and the extent of bypass flow, which was graded based on the findings of cerebral angiography, was investigated. RESULTS: Both the affected STA flow velocity and the STA flow velocity ratio, particularly those in the end diastole, increased in patients with more extensive bypass flow. In patients with extensive, moderate, and poor bypass flow, the end diastolic flow velocities of the operated STA were 27.4 +/- 8.8, 23.0 +/- 7.8, and 13.5 +/- 7.5 cm/s, respectively and the end diastolic flow velocity ratios of the STA were 3.4 +/- 0.8, 2.1 +/- 0.5 and 1.3 +/- 0.4, respectively. The pulsatility index and resistance index of the affected STA were significantly lower in the patients with more extensive bypass flow. The optimal threshold value of the end diastolic flow velocity ratio of STA for the group with extensive bypass flow was 2.75, whereas that for the group with poor bypass flow was 1.60. With the obtained values, the sensitivity and specificity were 87.5% and 93.9% for the group with extensive bypass flow and 95.2% and 95.0% for the group with poor bypass flow, respectively. CONCLUSION: The blood flow velocity in the operated STA seems to be a highly sensitive parameter for predicting the extent of bypass flow in patients undergoing STA-MCA anastomosis. PMID- 12748090 TI - Transarterial embolectomy in acute stroke. AB - BACKGROUND AND PURPOSE: An embolus causing cerebral ischemia is a major cause of death and disability, and the search for methods to reestablish blood flow is of major importance. A technique for the emergent primary treatment of cerebral emboli causing stroke is presented in detail. METHODS: The method developed implies the mechanical extraction of the embolus with a standard vascular retrieval snare via the endovascular route without the use of thrombolytic agents. RESULTS: Five consecutive patients were treated to extract an embolus. All patients had substantial improvement in their clinical status. No revascularization hemorrhages occurred. CONCLUSION: In this small series, the embolectomy method described was reproducible, rapid, and safe. It may offer an alternative to other methods of vascular recanalization. PMID- 12748091 TI - Temporal evolution of diffusion after spontaneous supratentorial intracranial hemorrhage. AB - BACKGROUND AND PURPOSE: The evolution of apparent diffusion coefficient abnormalities during supratentorial intracranial hemorrhage in normal appearing brain tissue has not been described. Recent investigations using diffusion imaging have revealed increased apparent diffusion coefficient in perihematomal tissue. We report brain tissue abnormalities beyond the visibly abnormal region ipsilateral and contralateral to the hematoma. This preliminary effort should generate meaningful clinical prognostic indicators for moderate size hemorrhages in large scale studies. METHODS: Using the neurology patient encounter database at a tertiary care hospital, we retrospectively identified patients who presented with acute focal neurologic deficits, had CT scans of the head that confirmed spontaneous intracranial hemorrhage, and had a MR images obtained within the first 6 hr to 30 days postictus. The regions identified as targets of this investigation were the hemorrhage and surrounding T2 signal intensity abnormality and the visibly normal supratentorial cerebral tissue. RESULTS: Ninety-five patients were admitted during a period of 25 months. Fifteen patients met the criteria for the study. Elevated whole brain diffusion was shown as early as 6 hr after intracranial hemorrhage. This increase in diffusion was comparable in both hemispheres. Diffusion values in the lesion (hematoma plus T2 signal intensity abnormality) increased slowly with peak increases noted 2 to 3 days after the ictus. CONCLUSION: Diffuse early cerebral response occurs in normal appearing brain tissue both ipsilateral and contralateral to the visibly abnormal hematoma, manifested by increased apparent diffusion coefficient. This response is present before the local response is fully developed. Supratentorial intracranial hemorrhage results in an early diffuse brain response with increased apparent diffusion coefficient in normal appearing brain. PMID- 12748092 TI - Parent vessel occlusion for vertebrobasilar fusiform and dissecting aneurysms. AB - BACKGROUND AND PURPOSE: Previous reports of outcome with permanent vessel occlusion (PVO) for large, giant, or fusiform aneurysms in the posterior circulation have been limited. We undertook this study to evaluate the perioperative (within 30 days) and follow-up outcomes for patients treated with permanent occlusion of the vertebral artery for vertebrobasilar fusiform and dissecting aneurysms. METHODS: Thirteen consecutive patients were studied. Two groups were defined for the study. Group I patients underwent PVO to achieve complete thrombosis of the aneurysm. Group II patients underwent PVO to reduce flow to the aneurysm where complete thrombosis was not desirable. Modified Rankin scores were obtained at presentation and at follow-up (follow-up range, 1-76 months; mean, 22.0 months). RESULTS: All group I aneurysms were shown to be thrombosed on the angiograms obtained at the immediate follow-up examinations. Improvement in outcome scores was achieved by all group I patients. Improvement in Rankin scores after endovascular treatment was statistically significant (P =.026). All group II patients had complete occlusion of the vertebral artery; however, continued filling of the fusiform aneurysm was still observed. Four patients in group II died during the follow-up period. Two of these deaths were attributable to the aneurysms. Of the remaining three patients, two experienced clinical worsening and one remained stable. CONCLUSION: In this series, PVO for chronic fusiform and acute dissecting aneurysms of the vertebrobasilar system proved to be a useful therapeutic endovascular technique. Long-term outcomes suggest that patients with aneurysms involving only one vertebral artery, where complete thrombosis can be achieved, have better clinical outcomes than those who have aneurysms involving the basilar artery or both vertebral arteries, where complete thrombosis cannot achieved by using PVO. PMID- 12748093 TI - Thalamic involvement in sporadic Creutzfeldt-Jakob disease: a diffusion-weighted MR imaging study. AB - BACKGROUND AND PURPOSE: Recent neuropathologic research suggests thalamic involvement in sporadic Creutzfeldt-Jakob disease (sCJD), which has been disregarded in imaging studies. Diffusion-weighted (DW) MR imaging has the highest sensitivity for the detection of signal intensity (SI) abnormalities in CJD. We hypothesized that pathologic changes in the thalamus in sCJD can be detected by using a subtle analysis of DW MR imaging. METHODS: Six sCJD patients and nine healthy controls were examined with a 1.5-T system by using DW single shot spin-echo echo planar (b = 0, 1000 s/mm(2)), T2-weighted turbo spin-echo, and fluid-attenuated inversion recovery sequences. One patient was examined serially (3, 4, and 8 months after onset of symptoms). MR images were reviewed for SI changes in the striatum, hippocampus, mediodorsal thalamic nucleus (MD), and pulvinar thalami. Apparent diffusion coefficients (ADCs) were measured in these areas. RESULTS: All sCJD patients showed increased SI on DW images in the striatum bilaterally. ADCs in these areas were significantly reduced. Four of six sCJD patients showed increased SI on DW images in the pulvinar thalami, whereas ADCs were significantly reduced in all patients (mean ADC +/- SEM: in patients with SI changes, 701 +/- 38; in patients without SI changes, 684 +/- 37; in controls, 853 +/- 15 [P <.0001]). No patient showed SI changes in the MD on DW images, whereas ADCs were significantly reduced in all (664 +/- 28 as compared with 800 +/- 24 in controls [P =.0011]). Serial measurements in one sCJD patient showed ADC reduction in the pulvinar thalami preceding the SI changes on DW images. CONCLUSION: A quantitative analysis of DW images with ADC measurements shows slight MR imaging changes in the thalamus in sCJD when abnormal SI may not be present. PMID- 12748094 TI - T1 hyperintensity in the pulvinar: key imaging feature for diagnosis of Fabry disease. AB - BACKGROUND AND PURPOSE: Fabry disease (FD) is an inborn error of glycosphingolipid metabolism. To date, no specific neuroimaging features have been elucidated to help in making the diagnosis of this disorder. The purpose of this study was to determine whether the finding of T1 shortening in the lateral pulvinar is a useful finding in the imaging diagnosis of FD and to deduce the relationship of this finding to the pathophysiology of the disease. METHODS: We studied T1- and T2-weighted images obtained in ten patients (nine male and one female) with FD with an age range of 19-59 years. The images were examined for anatomic aberrations and areas of abnormal signal intensity (SI) in both gray matter and white matter. The SI of deep gray matter was evaluated qualitatively and semiquantitatively, relative to the SI of CSF or the genu of the corpus callosum. Gradient echo MR images and axial noncontrast CT images were available for one patient. RESULTS: Seven of 10 patients showed small areas of T2 prolongation in the white matter of the cerebral hemispheres. Despite the known propensity for vascular disease in these patients, only one had cortical infarction. Bilateral T1 shortening in the lateral pulvinar was recognized in at least seven patients, all over the age of 30 years, who also had small areas of T2 prolongation in the white matter. CT and gradient echo images in one patient revealed no evidence of calcification or metallic deposits in the pulvinar. CONCLUSION: Bilateral T1 shortening in the lateral pulvinar is a common finding in FD and may be useful in suggesting this diagnosis. PMID- 12748095 TI - Clinical brain MR imaging prescriptions in Talairach space: technologist- and computer-driven methods. AB - BACKGROUND AND PURPOSE: Variability in patient head positioning may yield substantial interstudy image variance in the clinical setting. We describe and test three-step technologist and computer-automated algorithms designed to image the brain in a standard reference system and reduce variance. METHODS: Triple oblique axial images obtained parallel to the Talairach anterior commissure (AC) posterior commissure (PC) plane were reviewed in a prospective analysis of 126 consecutive patients. Requisite roll, yaw, and pitch correction, as three authors determined independently and subsequently by consensus, were compared with the technologists' actual graphical prescriptions and those generated by a novel computer automated three-step (CATS) program. Automated pitch determinations generated with Statistical Parametric Mapping '99 (SPM'99) were also compared. RESULTS: Requisite pitch correction (15.2 degrees +/- 10.2 degrees ) far exceeded that for roll (-0.6 degrees +/- 3.7 degrees ) and yaw (-0.9 degrees +/- 4.7 degrees ) in terms of magnitude and variance (P <.001). Technologist and computer generated prescriptions substantially reduced interpatient image variance with regard to roll (3.4 degrees and 3.9 degrees vs 13.5 degrees ), yaw (0.6 degrees and 2.5 degrees vs 22.3 degrees ), and pitch (28.6 degrees, 18.5 degrees with CATS, and 59.3 degrees with SPM'99 vs 104 degrees ). CATS performed worse than the technologists in yaw prescription, and it was equivalent in roll and pitch prescriptions. Talairach prescriptions better approximated standard CT canthomeatal angulations (9 degrees vs 24 degrees ) and provided more efficient brain coverage than that of routine axial imaging. CONCLUSION: Brain MR prescriptions corrected for direct roll, yaw, and Talairach AC-PC pitch can be readily achieved by trained technologists or automated computer algorithms. This ability will substantially reduce interpatient variance, allow better approximation of standard CT angulation, and yield more efficient brain coverage than that of routine clinical axial imaging. PMID- 12748096 TI - Abnormal perfusion of the pituitary gland secondary to dural arteriovenous fistulas in the cavernous sinus: dynamic MR findings. AB - BACKGROUND AND PURPOSE: If venous congestion is the primary cause of pituitary gland enlargement in cases of dural arteriovenous fistulas (AVFs), other abnormal pituitary findings may be detectable on MR images. We sought to investigate the perfusion abnormality of the pituitary gland secondary to dural AVFs in the cavernous sinus and to clarify its clinical importance. METHODS: Nine consecutive patients (all female; age range, 50-77 years) with dural AVFs in the cavernous sinus underwent prospective MR examinations, including dynamic studies, before and after therapy. Their clinical signs and symptoms were recorded. Two radiologists visually evaluated the enhancement patterns of the anterior pituitary gland. Dynamic MR curves were obtained by locating regions of interest at the center and bilateral peripheral areas of the anterior pituitary gland on coronal images. MR images obtained in five healthy individuals served as controls. RESULTS: No patient had symptoms of hypopituitarism or other endocrine abnormalities. Asymmetric pituitary enhancement was found in five patients; the side with the dural AVF was less enhancing. This finding disappeared after therapy. Although asymmetric enhancement was not detected in the remaining four patients, statistical analysis showed significantly delayed enhancement of the pituitary gland in the patients compared with enhancement patterns in control subjects. After treatment, this delay improved significantly. The pituitary gland significantly decreased in size after treatment. CONCLUSION: Perfusion of the pituitary gland is impaired in patients with a dural AVF in the cavernous sinus. This finding is probably due to venous congestion of the pituitary gland caused by high pressure in the cavernous sinus; it is usually not related to pituitary dysfunction. PMID- 12748097 TI - Peritumoral diffusion tensor imaging of high-grade gliomas and metastatic brain tumors. AB - BACKGROUND AND PURPOSE: Diffusion tensor imaging (DTI) is an advanced MR technique that describes the movement of water molecules by using two metrics, mean diffusivity (MD), and fractional anisotropy (FA), which represent the magnitude and directionality of water diffusion, respectively. We hypothesize that alterations in these values within the tissue surrounding brain tumors reflect combinations of increased water content and tumor infiltration and that these changes can be used to differentiate high-grade gliomas from metastatic lesions. METHODS: DTI was performed in 12 patients with high-grade gliomas and in 12 with metastatic lesions. DTI measurements were obtained from regions of interest (ROIs) placed on normal-appearing white matter and on the vasogenic edema, the T2 signal intensity abnormality surrounding each tumor. RESULTS: The peritumoral region of both gliomas and metastatic tumors displayed significant increases in MD (P <.005) and significant decreases in FA (P <.005) when compared with those of normal-appearing white matter. Furthermore, the peritumoral MD of metastatic lesions measured significantly greater than that of gliomas (P <.005). Peritumoral FA measurements, on the other hand, showed no such discrepancy. CONCLUSION: When compared with an internal control, diffusion metrics are clearly altered within the vasogenic edema surrounding both high-grade gliomas and metastatic tumors, reflecting increased extracellular water. Although peritumoral MD can be used to distinguish high-grade gliomas from metastatic tumors, peritumoral FA demonstrated no statistically significant difference. The FA changes surrounding gliomas, therefore, can be attributed not only to increased water content, but also to tumor infiltration. PMID- 12748099 TI - Proton MR spectroscopy of gliomatosis cerebri: case report of elevated myoinositol with normal choline levels. AB - A 69-year-old woman presented with clinical and imaging findings suspicious for gliomatosis cerebri, later confirmed by biopsy (moderately cellular, infiltrating glioma). Single voxel proton MR spectroscopy (TE 20 and TE 135) and spectroscopic imaging (TE 135) performed at admission showed normal choline, decreased N acetyl, and elevated myo-inositol levels relative to creatine. The primary conclusion is that in suspected cases of gliomatosis cerebri, myo inositol/creatine and myo-inositol/N-acetyl should be determined because they may provide evidence of tumor, even though choline/creatine is normal. A corollary to this conclusion is that choline/creatine may be misleading if used to demarcate infiltrating glioma from edema. PMID- 12748098 TI - Unusual MR spectroscopic imaging pattern of an astrocytoma: lack of elevated choline and high myo-inositol and glycine levels. AB - We present the case of a patient with an MR imaging study showing an ill-defined intra-axial mass in the right insula and frontal lobe. The mass showed high signal intensity on T2-weighted and fluid-attenuated inversion recovery images and an unusual proton MR spectroscopic imaging pattern characterized by the presence of high levels of myo-inositol/glycine, no significant elevation of choline, and mildly reduced N-acetylaspartate. The histopathologic diagnosis was of diffuse astrocytoma with oligodendroglial components (World Health Organization grade II). PMID- 12748100 TI - Proton MR spectroscopy in Wernicke encephalopathy. AB - Wernicke encephalopathy is caused by thiamine deficiency. Although the clinical picture has been well established for some time, clinical diagnosis is attained in only 20% of the cases. MR imaging techniques contribute to early diagnosis of Wernicke encephalopathy. We herein report MR imaging and proton MR spectroscopic findings for a patient with clinical and biochemical features consistent with Wernicke encephalopathy. Increased lactate and typical MR imaging findings are discussed in the context of the known pathophysiology of Wernicke encephalopathy. PMID- 12748101 TI - Basilar artery duplication associated with pituitary duplication: a new finding. AB - Pituitary duplication is a rare malformation, reported previously in approximately 18 patients. It is usually unsuspected before imaging, although it occurs most commonly in association with complicated midline and skull base anomalies. It is easily shown by MR imaging. Five new cases of pituitary duplication were diagnosed by using MR imaging studies reviewed at the Hospital for Sick Children. Among the many associated midline abnormalities, partial basilar artery duplication is a previously undescribed finding that we observed in all our cases. Cases of basilar artery duplication or fenestration are associated with altered flow dynamics, leading to a higher incidence of aneurysms. Periodic surveillance for this potential complication may be warranted. PMID- 12748102 TI - Demonstration of an actively bleeding aneurysm by CT angiography. AB - We describe a case of an actively bleeding aneurysm demonstrated by CT angiography (CTA). The active hemorrhage was characterized by a nebulous attenuation in the subarachnoid space adjacent to the aneurysm. The measurements (HU) of the hemorrhage were equal to or slightly less than those of the contrast material in the arteries and diminished as one moved further from the aneurysm. As CTA is used more frequently in the diagnosis of cerebral aneurysms, it is important to understand imaging features of this potentially lethal condition. PMID- 12748103 TI - Diffusion MR imaging changes associated with Wilson disease. AB - We herein report the case of a patient with Wilson disease. The patient underwent echo-planar diffusion MR imaging twice, 1.5 years apart. The lesions were in the putamina and caudate nuclei. At the first examination, undertaken after onset of extrapyramidal symptoms, a restricted diffusion pattern was evident. It is likely that this corresponded to cell swelling caused by the accumulation of copper. On the images obtained 1.5 years later, an opposite pattern (an elevated diffusion pattern) was noted. It is likely that this reflected necrosis, spongiform degeneration, and demyelination, which are among the known histopathologic changes associated with Wilson disease. PMID- 12748104 TI - A variant of the superficial middle cerebral vein mimicking an extraaxial hematoma. AB - We report a case of a 65-year-old man who was admitted to the hospital on an emergent basis because of head trauma. Non-contrast-enhanced CT findings of the brain suggested a small extraaxial hematoma that was shown to be a superficial middle cerebral vein variant on subsequent MR images. PMID- 12748105 TI - Varicella-zoster vasculitis presenting with intracranial hemorrhage. AB - Cerebral vasculitis presenting with intracranial hemorrhage is a rare but known entity. We discuss here the case of a 61-year-old woman presenting with subarachnoid hemorrhage. Cerebral angiography showed vasculitic changes involving the small and medium-sized vessels. She also had a concomitant herpes zoster rash on her back. A 3-month follow-up angiogram revealed partial resolution of the vasculitic changes. PMID- 12748106 TI - Preliminary experience with endovascular reconstruction for the management of carotid blowout syndrome. AB - BACKGROUND AND PURPOSE: Permanent balloon occlusion (PBO) of the carotid artery has been previously shown to be an effective means to treat carotid blowout syndrome (CBS). However, despite the effectiveness of this endovascular technique, concern remains regarding its potential for producing delayed cerebral ischemic complications in 15% to 20% of patients. This significant limitation of carotid PBO led our group to evaluate an alternative management strategy, consisting of endovascular reconstruction of the carotid artery (ERCA) in patients thought to be at particularly high risk for carotid occlusion (ie, provocative balloon test occlusion, angiographic documented incomplete circle of Willis, or contralateral carotid artery occlusion). METHODS: We reviewed all cases of CBS referred to our service, in which ERCA was chosen as a management strategy for patients thought to be at high risk for PBO, based on previously defined criteria. RESULTS: Sixteen carotid blowout events occurred in 12 patients with CBS who were deemed to be at high risk for cerebral ischemic complications, which were managed with ERCA by using a variety of stent devices and techniques. Adjunctive embolization of carotid pseudoaneurysms was performed in five of these patients by using platinum coils or acrylic glue. Hemostasis was achieved in all cases, although one patient with traumatic CBS and three patients with aggressive head and neck cancer-related CBS, required retreatment with ERCA. Recurrent CBS rates were similar to those reported in other studies using PBO. Overall, no treatment-related strokes or deaths occurred. CONCLUSION: CBS managed with ERCA can be performed safely and with efficacy of outcomes at least equivalent to those previously reported in association with conventional carotid PBO, therefore representing an excellent alternative endovascular technique for patients who are at increased risk of stroke after PBO. PMID- 12748107 TI - Guglielmi detachable coil extraction: suction generated by pusher-wire withdrawal after coil detachment within an intracranial aneurysm. AB - We present a case of Guglielmi detachable coil extraction into a microcatheter after coil detachment during embolization of a superior hypophyseal artery aneurysm; extraction was a result of the suction generated during pusher-wire withdrawal. Experimental simulations using many coil and microcatheter combinations were used to identify factors that contribute to the likelihood of this phenomenon. PMID- 12748108 TI - Treatment of acute intracranial vertebrobasilar dissection with angioplasty and stent placement: report of two cases. AB - Acute vertebrobasilar dissection may cause subarachnoid hemorrhage by rupturing through the adventia or cerebral infarct by progressive occlusion of the true lumen. Recent reports on the endovascular management of this condition have focused on treatment of pseudoaneurysms. We report two cases where angioplasty or stent placement was successfully used to improve compromised blood flow secondary to vertebrobasilar dissection. PMID- 12748109 TI - Coil embolization of an aneurysm associated with an infraoptic anterior cerebral artery in a child. AB - We report an unusual case of a ruptured aneurysm from an infraoptic anterior cerebral artery in an 11-year-old child. To our knowledge, this is the second such case treated with Guglielmi detachable coils in the pediatric population that has been described in the literature, excluding that of a vein of Galen aneurysm that resulted in remarkable recovery. We conclude that the endovascular technique is a feasible treatment option for children with ruptured cerebral aneurysms, especially when more complex vascular anomalies are present. PMID- 12748110 TI - Contrast-enhanced carotid color-coded duplex sonography for carotid stenting follow-up assessment. AB - Proper assessment of endovascular patency after carotid stent (CS) placement with carotid color-coded duplex sonography (CCCD) can be difficult. We investigated the usefulness of contrast-enhanced (CE) CCCD for post-CS follow-up. CCCD images could not depict the entire bloodstream in overlapped stents and in highly positioned stents. CE-CCCD images, however, did provide anatomic information almost equivalent to that of intra-arterial angiography. CE-CCCD is useful in screening for post-CS restenosis. PMID- 12748111 TI - Minimally invasive oxygen-ozone therapy for lumbar disk herniation. AB - BACKGROUND AND PURPOSE: Oxygen-ozone therapy is a minimally invasive treatment for lumbar disk herniation that exploits the biochemical properties of a gas mixture of oxygen and ozone. We assessed the therapeutic outcome of oxygen-ozone therapy and compared the outcome of administering medical ozone alone with the outcome of medical ozone followed by injection of a corticosteroid and an anesthetic at the same session. METHODS: Six hundred patients were treated with a single session of oxygen-ozone therapy. All presented with clinical signs of lumbar disk nerve root compression, with CT and/or MR evidence of contained disk herniation. Three hundred patients (group A) received an intradiscal (4 mL) and periganglionic (8 mL) injection of an oxygen-ozone mixture at an ozone concentration of 27 micro g/mL. The other 300 patients (group B) received, in addition, a periganglionic injection of corticosteroid and anesthetic. Therapeutic outcome was assessed 6 months after treatment by using a modified MacNab method. Results were evaluated by two observers blinded to patient distribution within the two groups. RESULTS: A satisfactory therapeutic outcome was obtained in both groups. In group A, treatment was a success (excellent or good outcome) in 70.3% and deemed a failure (poor outcome or recourse to surgery) in the remaining 29.7%. In group B, treatment was a success in 78.3% and deemed a failure in the remaining 21.7%. The difference in outcome between the two groups was statistically significant (P <.05). CONCLUSION: Combined intradiscal and periganglionic injection of medical ozone and periganglionic injection of steroids has a cumulative effect that enhances the overall outcome of treatment for pain caused by disk herniation. Oxygen-ozone therapy is a useful treatment for lumbar disk herniation that has failed to respond to conservative management. PMID- 12748112 TI - Intrathecal injection of contrast medium to prevent polymethylmethacrylate leakage during percutaneous vertebroplasty. AB - The major technical drawback of percutaneous vertebroplasty is the potential for neural compromise from leakage of polymethylmethaorylate into epidural or perivertebral veins. We have combined the procedure of intrathecal injection of contrast medium with vertebroplasty to better delineate spinal canal encroachment during injection when the posterior vertebral wall is compromised by myeloma. PMID- 12748113 TI - Sacroplasty: a treatment for sacral insufficiency fractures. AB - Sacral insufficiency fracture is a debilitating injury for which no active treatment is currently available. We present three consecutive cases of elderly patients with sacral insufficiency fractures whose symptoms were relieved immediately by treatment of the fractures by polymethylmethacrylate injections, a so-called sacroplasty. PMID- 12748114 TI - MR imaging of an infiltrating spinal epidural angiolipoma. AB - Infiltrating spinal epidural angiolipoma is an uncommon benign tumor composed of mature adipose elements admixed with abnormal blood vessel, which tends to invade the surrounding soft tissue and may potentially be mistaken for an aggressive tumor. In this report, we present the MR imaging findings of a pathologically proved infiltrating spinal epidural angiolipoma that appeared largely hypointense on T1-weighted images and enhanced strongly with IV injection of contrast medium, features that suggested a malignant tumor. PMID- 12748116 TI - Forget About "van der Knaap syndrome," forget about glycine. PMID- 12748115 TI - Prospective evaluation of carotid artery stenosis: elliptic centric contrast enhanced MR angiography and spiral CT angiography compared with digital subtraction angiography. AB - BACKGROUND AND PURPOSE: Although digital subtraction angiography (DSA) is the reference standard for assessing carotid arteries, it is uncomfortable for patients and has a small risk of disabling stroke and death. These problems have fueled the use of spiral CT angiography and MR angiography. We prospectively compared elliptic centric contrast-enhanced MR angiography and spiral CT angiography with conventional DSA for detecting carotid artery stenosis. METHODS: Eighty carotid arteries (in 40 symptomatic patients) were assessed. Elliptic centric MR and spiral CT angiographic data were reconstructed with maximum intensity projection and multiplanar reconstruction techniques. All patients had been referred for DSA evaluation on the basis of findings at Doppler sonography, which served as a screening method (degree of stenosis > or = 70% or inconclusive results). Degree of carotid stenosis estimated by using the three modalities was compared. RESULTS: Significant correlation with DSA was found for stenosis degree for both elliptic centric MR and spiral CT angiography; however, the correlation coefficient was higher for MR than for CT angiography (r = 0.98 vs r = 0.86). Underestimation of stenoses of 70-99% occurred in one case with elliptic centric MR angiography (a 70% stenosis was underestimated as 65%) and in nine cases with spiral CT angiography, in comparison to DSA findings. Overestimation occurred in two cases with MR angiography (stenoses of 65-67% were overestimated as 70-75%). With CT, overestimation occurred in seven cases; a stenosis of 60% in one case was overestimated as 70%. Both techniques confirmed the three cases of carotid occlusion. With elliptic centric MR angiography, carotid stenoses of 70% or greater were detected with high sensitivity, 97.1%; specificity, 95.2%; likelihood ratio (LR) for a positive test result, 20.4; and ratio of LR(+) to LR( ), -0.3. With spiral CT angiography, sensitivity, specificity, LR(+), and LR(+):LR(-) were 74.3%, 97.6%, 31.2, and 0.3, respectively. CONCLUSION: Elliptic centric contrast-enhanced MR angiography is more accurate than spiral CT angiography to adequately evaluate carotid stenosis. Furthermore, elliptic centric contrast-enhanced MR angiography appears to be adequate to replace conventional DSA in most patients examined. PMID- 12748117 TI - PDC-109 (BSP-A1/A2) promotes bull sperm binding to oviductal epithelium in vitro and may be involved in forming the oviductal sperm reservoir. AB - Sperm reservoirs have been found in the oviducts of several species of mammals. In cattle, the reservoir is formed by the binding of sperm to fucose-containing glycoconjugates on the surface of oviductal epithelial cells. A fucose-binding molecule was purified from sperm extracts and identified as PDC-109 (BSP-A1/A2), a protein that is secreted by the seminal vesicles and associates with the plasma membrane of sperm upon ejaculation. The objective of this study was to demonstrate that PDC-109 promotes bull sperm binding to oviductal epithelium. PDC 109 was purified from bovine seminal plasma, and polyclonal antibodies were produced in rabbits. The antibodies detected PDC-109 on ejaculated sperm by indirect immunofluorescence and Western blots of extracts, but PDC-109 was not detected on epididymal sperm. When added to epididymal sperm, purified PDC-109 was absorbed onto the plasma membrane overlying the acrosome, as demonstrated by indirect immunofluorescence and by labeling sperm directly with fluorescein conjugated PDC-109. When added to explants of oviductal epithelium, significantly fewer epididymal sperm than ejaculated sperm became bound. Addition of PDC-109 to epididymal sperm increased epithelial binding to the level observed for ejaculated sperm. In addition, binding of ejaculated sperm to oviductal epithelium was inhibited by addition of excess soluble PDC-109. Ejaculated sperm lost the ability to bind to oviductal epithelium after heparin-induced capacitation, but treatment with PDC-109 restored binding. These results demonstrate that PDC-109 enables sperm to bind to oviductal epithelium and plays a major role in formation of the bovine oviductal sperm reservoir. PMID- 12748118 TI - Photoperiod and testosterone regulate androgen receptor immunostaining in the Siberian hamster brain. AB - Day length regulates the effects of gonadal steroids on gonadotropin secretion and behavior in seasonal breeders. To determine whether this influence of photoperiod results from changes in androgen receptor expression in Siberian hamster brain regions that regulate neuroendocrine function, androgen receptor immunostaining was examined in castrated animals given either no androgen replacement or one of three doses of testosterone (T) resulting in physiological serum concentrations. Half of the animals were housed under inhibitory photoperiod conditions, and immunostaining was quantified 11 days later. Measurement of serum gonadotropin and prolactin concentrations confirmed that androgen exerted graded effects on pituitary function but that the animals were killed before photoperiodic influences had fully developed. T significantly increased the numbers of androgen receptor-immunoreactive cells in every brain region examined. Photoperiod exerted no significant influence on androgen receptor-immunoreactive cell number in the arcuate nucleus, bed nucleus of the stria terminalis (BNST), medial preoptic nucleus, or in medial amygdala. An interaction between T and photoperiod was observed in the BNST and in the rostral and middle portions of the arcuate nucleus. Although increasing concentrations of T resulted in more intense cellular immunostaining in the BNST and arcuate, this effect was not influenced by day length. These results indicate that relatively short-duration (11 days) exposure to inhibitory photoperiod triggers localized and regionally specific changes in androgen receptor expression. PMID- 12748120 TI - The activin-follistatin system in the neonatal ovine uterus. AB - Uterine gland development or adenogenesis in the neonatal ovine uterus involves budding and tubulogenesis followed by coiling and branching morphogenesis of the glandular epithelium (GE) from the luminal epithelium (LE) between birth (Postnatal Day [PND] 0) and PND 56. Activins, which are members of the transforming growth factor beta superfamily, and follistatin, an inhibitor of activins, regulate epithelial branching morphogenesis in other organs. The objective of the present study was to determine effects of postnatal age on expression of follistatin, inhibin alpha subunit, betaA subunit, betaB subunit, activin receptor (ActR) type IA, ActRIB, and ActRII in the developing ovine uterus. Ewes were ovariohysterectomized on PND 0, 7, 14, 21, 28, 35, 42, 49, or 56. The uterus was analyzed by in situ hybridization and immunohistochemistry. Neither inhibin alpha subunit mRNA or protein was detected in the neonatal uterus. Expression of betaA and betaB subunits was detected predominantly in the endometrial LE and GE and myometrium between PND 0 and PND 56. In all uterine cell types, ActRIA, ActRIB, and ActRII were expressed, with the highest levels observed in the endometrial LE and GE and myometrium. Between PND 0 and PND 14, follistatin was detected in all uterine cell types. However, between PND 21 and PND 56, follistatin was only detected in the stroma and myometrium and not in the developing GE. Collectively, the present results indicate that components of the activin-follistatin system are expressed in the developing neonatal ovine uterus and are potential regulators of endometrial gland morphogenesis. PMID- 12748119 TI - Altered Sonic hedgehog signaling is associated with morphological abnormalities in the penis of the BB/WOR diabetic rat. AB - Erectile dysfunction (ED) is a common and debilitating pathological development that affects up to 75% of diabetic males. Neural stimulation is a crucial aspect of the normal erection process. Nerve injury causes ED and disrupts signaling of the Sonic hedgehog (Shh) cascade in the smooth muscle of the corpora cavernosa. Shh and targets of its signaling establish normal corpora cavernosal morphology during postnatal differentiation of the penis and regulate homeostasis in the adult. Interruption of the Shh cascade in the smooth muscle of the corpora cavernosa results in extensive changes in corpora cavernosal morphology that lead to ED. Our hypothesis is that the neuropathy observed in diabetics causes morphological changes in the corpora cavernosa of the penis that result in ED. Disruption of the Shh cascade may be involved in this process. We tested this hypothesis by examining morphological changes in the penis, altered gene and protein expression, apoptosis, and bromodeoxyuridine incorporation in the BB/WOR rat model of diabetes. Extensive smooth muscle and endothelial degradation was observed in the corpora cavernosa of diabetic penes. This degradation accompanied profound ED, significantly decreased Shh protein in the smooth muscle of the corpora cavernosa, and increased penile Shh RNA expression in the intact penis (nerves, corpora, and urethra). Localization and expression of Shh targets were also disrupted in the corpora cavernosa. Increasing our understanding of the molecular mechanisms that regulate Shh signaling may provide valuable insight into improving treatment options for diabetic impotence. PMID- 12748122 TI - Oviduct-specific glycoprotein modulates sperm-zona binding and improves efficiency of porcine fertilization in vitro. AB - Oviduct-specific glycoprotein (OGP) displays estrus-associated regional and temporal differences in expression and localizes to the zona pellucida, perivitelline space, and plasma membrane of oviductal oocytes and embryos, suggesting that it may have a role in regulation of fertilization and/or early embryonic development. The aims of this study were to evaluate the effect of exogenous OGP on in vitro fertilization (IVF) and embryo development in the pig using a defined serum-free culture system. In vitro-matured porcine oocytes were incubated with homologous OGP (0, 1, 10, 20, and 40 microg/ml) for 3 h and then washed prior to IVF. Exposure of oocytes to 10 or 20 microg/ml porcine OGP (pOGP) significantly reduced the incidence of polyspermy compared with the control (P < 0.01) while maintaining high penetration rates. When oocytes, spermatozoa, or both were preincubated with 10 microg/ml pOGP prior to IVF, the incidence of polyspermy was similarly reduced (P < 0.01) by all three treatments without affecting penetration rates. The ability of spermatozoa to undergo calcium ionophore-induced acrosome reaction was similar with or without exposure to pOGP. However, significantly fewer spermatozoa (P < 0.01) bound to the zona pellucida when oocytes were preincubated with pOGP. To evaluate the effect of pOGP on embryo development, embryos were cultured in pOGP-supplemented medium for 48 h or 144 h. Both transient and continuous exposure to pOGP significantly enhanced cleavage and blastocyst formation rate compared with the control (P < 0.01). These data demonstrate that exposure of either in vitro-matured oocytes or spermatozoa to pOGP decreased polyspermy and spermatozoa binding while maintaining high penetration rates of pig oocytes fertilized in vitro. Furthermore, pOGP exerted an embryotrophic effect independent of effects demonstrated on spermatozoa and oocytes at fertilization. PMID- 12748121 TI - Ovarian regulation of endometrial gland morphogenesis and activin-follistatin system in the neonatal ovine uterus. AB - Postnatal development of the ovine uterus between birth and Postnatal Day (PND) 56 involves differentiation of the endometrial glandular epithelium from the luminal epithelium followed by tubulogenesis and branching morphogenesis. Previous results indicated that ovariectomy of ewes at birth did not affect uterine growth or initial stages of endometrial gland genesis on PND 14 but did affect uterine growth after PND 28. Available evidence from a number of species supports the hypothesis that the ovary does not affect endometrial gland morphogenesis in the postnatal uterus. To test this hypothesis in our sheep model, ewes were assigned at birth to a sham surgery as a control or bilateral ovariectomy (OVX) on PND 7. Uteri were removed and weighed on PND 56. Ovariectomy did not affect circulating levels of estradiol-17beta. Uterine weight was 52% lower in OVX ewes. Histomorphological analyses indicated that the thickness of the endometrium and myometrium, total number of endometrial glands, and endometrial gland density in the stratum spongiosum stroma was reduced in uteri of OVX ewes. In contrast, the number of superficial ductal gland invaginations and gland density in the stratum compactum stroma was not affected by ovariectomy. The uteri of OVX ewes contained lower levels of betaA subunit, activin receptor (ActR) type IA, ActRIB, and follistatin protein expression but higher levels of betaB subunit. In the neonatal ovary, follistatin, inhibin alpha subunit, betaA subunit, and betaB subunit were expressed in antral follicles between PNDs 0 and 56. These results led to rejection of the hypothesis that the ovary does not influence endometrial adenogenesis. Rather, the ovary and, thus, an ovarian-derived factor regulates, in part, the coiling and branching morphogenetic stage of endometrial gland development after PND 14 and expression of specific components of the activin-follistatin system in the neonatal ovine uterus that appear to be important for that critical process. PMID- 12748123 TI - Identification of developmentally regulated genes in the somatic cells of the mouse testis using serial analysis of gene expression. AB - To identify genes developmentally regulated in the somatic cells of the testis, serial analysis of gene expression (SAGE) has been used to generate gene expression profiles from these cells in the fetal and adult mouse. To avoid germ cell transcripts, a fetal SAGE library was generated from germ cell-free fetal Wv/Wv mice, and an adult SAGE library was generated from adult testes depleted of germ cells with busulfan. The combined SAGE libraries contained 147570 tags identifying 12976 unique transcripts. Of these transcripts, 3607 were present in only the fetal library and 3941 were present in only the adult library. Most of the abundant differentially expressed tags in the adult testis library were from characterized genes, whereas 3' rapid amplification of complementary ends was required to identify most differentially expressed tags in the fetal library. These fetal tags were mostly associated with uncharacterized UniGene clusters. These data provide a comprehensive and quantitative analysis of gene expression in the somatic cells of the fetal and adult testis (including unknown transcripts) and identify genes differentially expressed in these cells during testis development. These differentially regulated genes are likely to provide insight into mechanisms regulating testis function both during development and in the adult animal. PMID- 12748124 TI - Class II transactivator (CIITA) promoter methylation does not correlate with silencing of CIITA transcription in trophoblasts. AB - Trophoblast cells are unique because they do not express major histocompatibility complex (MHC) class II antigens, either constitutively or after exposure to interferon-gamma (IFN-gamma). The absence of MHC class II antigens on trophoblasts is thought to play a critical role in preventing rejection of the fetus by the maternal immune system. The inability of trophoblasts to express MHC class II genes is primarily due to lack of the class II transactivator (CIITA), a transacting factor that is required for constitutive and IFN-gamma-inducible MHC class II transcription. We, therefore, investigated the silencing of CIITA expression in trophoblasts. In transient transfection assays, transcription from the IFN-gamma-responsive CIITA type IV promoter was upregulated by IFN-gamma in trophoblasts, which suggests that CIITA is silenced by an epigenetic mechanism in these cells. Polymerase chain reaction analysis demonstrated that the CIITA type IV promoter is methylated in both the human choriocarcinoma cell lines JEG-3 and Jar and in 2fTGH fibrosarcoma cells, which are IFN-gamma inducible for CIITA. Conversely, methylation of the CIITA type IV promoter was not observed in human primary cytotrophoblasts isolated from term placentae or in mouse or rat trophoblast cell lines. Simultaneous treatment with IFN-gamma and the histone deacetylase inhibitor trichostatin A weakly activated CIITA transcription in mouse trophoblasts. Stable hybrids between human choriocarcinoma and fibrosarcoma cells and between mouse trophoblasts and fibroblasts expressed CIITA following treatment with IFN-gamma. These results suggest that silencing of CIITA transcription is recessive in trophoblasts and involves an epigenetic mechanism other than promoter methylation. The fact that CIITA is expressed in the stable hybrids implies that trophoblasts may be missing a factor that regulates chromatin structure at the CIITA promoter. PMID- 12748125 TI - Disruption of imprinted gene methylation and expression in cloned preimplantation stage mouse embryos. AB - Cloning by somatic cell nuclear transfer requires that epigenetic information possessed by the donor nucleus be reprogrammed to an embryonic state. Little is known, however, about this remodeling process, including when it occurs, its efficiency, and how well epigenetic markings characteristic of normal development are maintained. Examining the fate of epigenetic information associated with imprinted genes during clonal development offers one means of addressing these questions. We examined transcript abundance, allele specificity of imprinted gene expression, and parental allele-specific DNA methylation in cloned mouse blastocysts. Striking disruptions were seen in total transcript abundance and allele specificity of expression for five imprinted genes. Only 4% of clones recapitulated a blastocyst mode of expression for all five genes. Cloned embryos also exhibited extensive loss of allele-specific DNA methylation at the imprinting control regions of the H19 and Snprn genes. Thus, epigenetic errors arise very early in clonal development in the majority of embryos, indicating that reprogramming is inefficient and that some epigenetic information may be lost. PMID- 12748127 TI - Male mice lacking the Theg (testicular haploid expressed gene) protein undergo normal spermatogenesis and are fertile. AB - The testicular haploid expressed gene (Theg) encodes for a novel approximately 42.0-kDa nuclear protein, which is specifically expressed in spermatid cells. Its expression is upregulated by some unknown factor(s) from Sertoli cells. To elucidate the function of Theg protein and its role in spermatogenesis, we disrupted the Theg locus in mouse by homologous recombination. For functional dissection of the domain structure of the Theg protein, two different knockout approaches were undertaken. In the first knockout mouse (Th14), the C-terminal region of the Theg protein (amino acids 137-376) was deleted. Both Th14+/- and Th14-/- mice from genetic backgrounds of C57BL/6J x 129X1/SvJ hybrid and 129X1/SvJ inbred exhibited a normal phenotype and were fertile. The testes of Th14-/- mice were smaller than those of Th14+/- and Th14+/+ mice; however, the testicular morphology and the properties of sperm, including morphology and motility, from Th14-/- mice were similar to those of Th14+/- and Th14+/+ mice. These results demonstrate that the C-terminal region of Theg (amino acids 137 376) does not play an important role in progression of spermatogenesis. In the second knockout mouse (Th15), we deleted the N-terminal domain of the Theg protein, which resulted in complete loss of Theg transcripts. Both Th15+/- and Th15-/- mice from genetic backgrounds C57BL/6J x 129X1/SvJ hybrid, C3H/J congenic, and 129X1/SvJ inbred appeared normal and were fertile, with no gross abnormalities detected in testicular morphology or sperm properties. Our results from both knockout mouse model systems clearly illustrate that Theg is not essential for spermatogenesis in the mouse. PMID- 12748126 TI - Gestational exposure to ethane dimethanesulfonate permanently alters reproductive competence in the CD-1 mouse. AB - Although the adult mouse Leydig cell (LC) has been considered refractory to cytotoxic destruction by ethane dimethanesulfonate (EDS), the potential consequences of exposure during reproductive development in this species are unknown. Herein pregnant CD-1 mice were treated with 160 mg/kg on Gestation Days 11-17, and reproductive development in male offspring was evaluated. Prenatal administration of EDS compromised fetal testosterone (T) levels, compared with controls. EDS-exposed pups recovered their steroidogenic capacities after birth because T production by hCG-stimulated testis parenchyma from prepubertal male offspring was unchanged. However, prepubertal testes from prenatally exposed males contained seminiferous tubules (STs) devoid of germ cells, indicating a delay in spermatogenesis. In adults, some STs in exposed males still contained incomplete germ cell associations corroborating observed reductions in epididymal sperm reserves, fertility ratios, and litter size. Morphometry revealed an EDS induced increase in interstitial area and a concomitant decrease in ST area, but stereology revealed an unexpected decrease in the number and size of the LCs per testis in exposed males. Paradoxically, there was an increase in both serum LH and T production by adult testis parenchyma, indicating that the LCs were hyperstimulated. These data demonstrate permanent lesions in LC development and spermatogenesis caused by prenatal exposure in mice. Thus, although adult mouse LCs are insensitive to EDS, EDS appears to have direct action on fetal LCs, resulting in abnormal testis development. PMID- 12748128 TI - Characterization of ovarian follicular wave dynamics in women. AB - A wave phenomenon of ovarian follicular development in women has recently been documented in our laboratory. The objective of the present study was to characterize follicular waves to determine whether women exhibit major and minor wave patterns of follicle development during the interovulatory interval (IOI). The ovaries of 50 women with clinically normal menstrual cycles were examined daily using transvaginal ultrasonography for one IOI. Profiles of the diameters of all follicles >or=4 mm and the numbers of follicles >or=5 mm were graphed during the IOI. Major waves were defined as those in which one follicle grew to >or=10 mm and exceeded all other follicles by >or=2 mm. Minor waves were defined as those in which follicles developed to a diameter of <10 mm and follicle dominance was not manifest. Blood samples were drawn to measure serum concentrations of estradiol-17beta, LH, and FSH. Women exhibited major and minor patterns of follicular wave dynamics during the IOI. Of the 50 women evaluated, 29/34 women with two follicle waves (85.3%) exhibited a minor-major wave pattern of follicle development and 5 women (14.7%) exhibited a major-major wave pattern. Ten of the 16 women with three follicle waves (62.5%) exhibited a minor-minor major wave pattern, 3 women (18.8%) exhibited a minor-major-major wave pattern, and 3 women (18.8%) exhibited a major-major-major wave pattern. Documentation of major and minor follicular waves during the menstrual cycle challenges the traditional theory that a single cohort of antral follicles grows only during the follicular phase of the menstrual cycle. PMID- 12748129 TI - Epigenetic characteristics and development of embryos cloned from donor cells treated by trichostatin A or 5-aza-2'-deoxycytidine. AB - Development to blastocyst following nuclear transfer is dependent on the donor cell's ability to reprogram its genome to that of a zygote. This reprogramming step is inefficient and may be dependent on a number of factors, including chromatin organization. Trichostatin A (TSA; 0-5 microM), a histone deacetylase inhibitor, was used to increase histone acetylation and 5-aza-2'-deoxycytidine (5 aza-dC; 0-5 microM), a DNA methyl-transferase inhibitor, was used to decrease methylation of chromatin in donor cells in an attempt to improve their reprogrammability. Adult fibroblast cells treated with 1.25 or 5 microM TSA had elevated histone H3 acetylation compared to untreated controls. Cells treated with 0.3 microM 5-aza-dC had decreased methylation compared to untreated controls. Both drugs at 0.08 microM caused morphological changes of the donor cells. Development to blastocysts by embryos cloned from donor cells after 0.08 or 0.3 microM 5-aza-dC treatments was lower than in embryos cloned from untreated control cells (9.7% and 4.2%, respectively, vs. 25.1%), whereas 0.08 microM TSA treatment of donor cells increased blastocyst development compared to controls (35.1% vs. 25.1%). These results indicate that partial erasure of preexisting epigenetic marks of donor cells improves subsequent in vitro development of cloned embryos. PMID- 12748130 TI - Emergency management of diabetic ketoacidosis in adults. AB - The authors propose a regimen for managing diabetic ketoacidosis in adults based on available evidence and their experience in the emergency department. PMID- 12748131 TI - Nitrous oxide in emergency medicine. AB - Safe and predictable analgesia is required for the potentially painful or uncomfortable procedures often undertaken in an emergency department. The characteristics of an ideal analgesic agent are safety, predictability, non invasive delivery, freedom from side effects, simplicity of use, and a rapid onset and offset. Newer approaches have threatened the widespread use of nitrous oxide, but despite its long history this simple gas still has much to offer. "I am sure the air in heaven must be this wonder-working gas of delight". Robert Southey, Poet (1774 to 1843) PMID- 12748132 TI - Randomised controlled trial of single, subacromial injection of methylprednisolone in patients with persistent, post-traumatic impingment of the shoulder. AB - OBJECTIVE: To evaluate the impact on recovery, of single subacromial injection of methylprednisolone in patients with persistent, post-traumatic impingement of the shoulder. DESIGN: Randomised, controlled study. SETTING: Large accident and emergency department in Leicester, UK. PARTICIPANTS: 98 patients with persistent, post-traumatic impingement of the shoulder. INTERVENTION: Single subacromial injection of methylprednisolone with bupivicaine (group S, n=54) or bupivicaine only (group C, n=44). MAIN OUTCOME MEASURES: Pain using a 10 cm visual analogue scale (VAS) and active shoulder abduction. RESULTS: Comparison of pain scores by the 10 cm VAS between group and group C showed no statistical difference at 3, 6, or 12 weeks. Mean patient pain scores at 12 weeks were 1.38 in both groups (p=0.99). There were 16 patients in group S (mean age 52 years) with a 10 cm VAS greater than 1 (95% CI CI 0.17 to 0.43), compared with 13 patients (mean age 57 years) in group C (95% CI 0.17 to 0.45). Comparison of active shoulder abduction between group S and group C showed no statistical difference at 3, 6, or 12 weeks. Mean active abduction at 12 weeks was 168.9 degrees in group S and 170.3 degrees in group C (p=0.8). There were 10 patients in group S (mean age 60.5) with active abduction less than 170 at 12 weeks (95% CI 0.09 to 0.31), compared with five patients (mean age 62 years) in group C (95% CI 0.04 to 0.24). CONCLUSIONS: Single subacromial injection of methylprednisolone has no beneficial impact on reducing the pain, or the duration of immobility in patients with persistent post-traumatic impingement of the shoulder PMID- 12748133 TI - National audit of emergency department child protection procedures. AB - OBJECTIVE: To assess the compliance with national guidelines on child protection procedures and provision of paediatric services in major English emergency departments. BACKGROUND: Victims of child abuse may present to emergency departments, and successful detection and management depends on adequate child protection procedures being in place. Two official documents published in 1999 provide recommendations for child protection procedures and staffing arrangements in emergency departments, and these can be used as standards for audit. METHODS: Structured telephone questionnaire survey of English emergency departments receiving at least 18 000 child attenders per year. RESULTS: Many of the standards are being met. Areas for improvement include: better access to child protection registers with clearer indications for their use; improved communication with other professionals such as the school nurse; more formal training for medical and nursing staff in the identification of potential indicators of child abuse; and improved awareness of local named professionals with expertise in child protection. More consultants with training in paediatric emergency medicine and more registered children's nurses are needed. CONCLUSION: Many nationally agreed recommendations are being met, but there is a need for improved training, increased numbers of specialised staff, and improved communication between professionals. There is considerable variation in practice between departments. PMID- 12748134 TI - Cervical spine injuries to children under 11: should we use radiography more selectively in their initial assessment? AB - OBJECTIVES: To assess the effectiveness of cervical spine radiography in injured children under 11 years old, and suggest improvements. METHODS: Retrospective survey of radiographs and accident and emergency records for children examined during a one year period in a large teaching hospital. RESULTS: No cervical spine fractures occurred in this age group during the year. The recorded clinical findings did not always justify radiography. CONCLUSIONS: Clinical examination appears undervalued by those assessing injured children and is poorly recorded. Radiography can be used more selectively. Initial assessment using a single lateral projection can be followed in doubtful cases by cross sectional imaging. PMID- 12748135 TI - "Why haven't you taken any pain killers?" A patient focused study of the walking wounded in an urban emergency department. AB - OBJECTIVES: (1) To assess the proportion of patients of triage category 3-5 presenting to the minor side of an urban emergency department who present without taking prior pain relief, and (2) to describe the reasons why they do not take pain relief for their presenting complaint METHOD: By patient interview of a convenience sample of 60 adult patients in the setting of an urban emergency department. RESULTS: Fifteen of 60 patients had taken analgesia and 45 of 60 (75%) had not. Sixteen reasons were volunteered to the interviewer. Most patients offered one reason only 39 of 45 (87%). The three commonest single reasons cited for not taking pain relief were "don't like taking tablets" 10 (22%), "run out of tablets" 10 (22%), five (11%) said their "pain not bad enough". Six (13%) patients cited two reasons for not taking pain relief. Only three (6%) patients indicated that they "did not think about pain relief". Six (13%) of patients had inappropriate perceptions of how pain killers may interfere with their care. CONCLUSION: Most patients presenting with painful conditions to the minor side of an urban emergency department had not taken pain relief. The study highlights there are many different reasons for this and staff should not presume that it was because the patient "did not think about it". Ongoing education of staff and patients is needed. PMID- 12748136 TI - Stress and coping in accident and emergency senior house officers. AB - OBJECTIVES: To identify levels of psychological distress in accident and emergency (A&E) senior house officers (SHOs). METHODS: Questionnaire survey given to SHOs at training sessions. MEASURES: General Health Questionnaire (GHQ) and the Brief COPE. RESULTS: Over half of the 37 respondents (51%; 95% CI: 36% to 67%) scored over the threshold for psychological distress on the GHQ. This is considerably higher than for other groups of doctors and for other professional groups. Correlational analysis revealed that the coping style Venting was significantly related to greater anxiety (r=0.34; p<0.05) and depression (r=0.33; p<0.05), while the coping style Active was significantly related to lower anxiety (r=-0.38; p<0.05), somatic complaints (r=-0.46; p<0.001) and years since qualification (r=0.40; p<0.05). CONCLUSIONS: Replicating findings from a study of stress in A&E consultants, this study shows higher levels of overt psychological distress among A&E SHOs than among other groups of doctors. An intervention to improve coping strategies may be useful for this group of doctors. PMID- 12748137 TI - Use of, and outputs from, an assault patient questionnaire within accident and emergency departments on Merseyside. AB - OBJECTIVES: To describe the implementation, use of, and outputs from an assault patient questionnaire (APQ) introduced in accident and emergency (A&E) departments to determine Crime & Disorder and Community Safety priorities on Merseyside, a metropolitan county in north west England, UK. METHODS: Why and how the APQ was implemented, data collected, and information obtained. The subsequent incorporation of the APQ into the Torex Patient Administration System (PAS) at the Royal Liverpool University Hospital A&E department and its routine completion by trained reception staff. RESULTS: Analysis is based upon anonymised data-for example, patient ID and date of birth information is suppressed. A summary of "baseline" information obtained from the data collected is provided. CONCLUSIONS: It is possible for the APQ to be implemented at no extra cost in a large A&E department in an acute general teaching hospital. Valuable intelligence can be obtained for Crime & Disorder Act and Community Safety processes. The APQ forms part of a medium to long term strategy to prevent and reduce violent assaults in the community that subsequently require treatment in an A&E department. Such incidents include assaults both inside and outside licensed premises, attacks by strangers on the street, and domestic violence. Emphasis is also placed upon the feedback of results to staff in A&E departments. PMID- 12748138 TI - How to get on a SpR rotation in emergency medicine (and make the most of it). AB - Emergency medicine is now proving a popular specialty in the United Kingdom. A recent report ranks emergency medicine second in specialties attracting the most applications for specialist registrar (SpR) interview. Numbered posts are becoming increasingly competitive as a result. This paper offers advice to aspiring emergency department SpRs. It identifies areas in which a curriculum vitae may be improved. It should also enable emergency department trainees to set objectives for their early SpR years. PMID- 12748139 TI - Descriptive epidemiology of adult critical care transfers from the emergency department. AB - AIMS: To describe the nature, frequency, and characteristics of adult critical care transfers originating from the emergency department (ED). METHODS: A one year prospective regional descriptive study using multiple data sources of all critically ill adults transferred from an ED or a minor injuries unit (MIU) within the former Yorkshire Regional Health Authority Area or into a regional critical care facility if originating from an ED or MIU elsewhere. RESULTS: 29 EDs transferred 349 adults into the regional critical care facilities. The median number of transfers per department within the region was 18 (range 1 to 42). Seventeen were transferred from outside the region. A total of 263 (75%) patients were transferred for specialist care and 76 (22%) for non-clinical reasons. Altogether 294 (84%) were admitted to intensive care or a high dependency unit at the receiving hospital. The in-hospital documented mortality rate was 26%. A total of 170 patients (49%) had traumatic pathology of which 101 were principally transferred for management of a head injury. Median time in the ED was 3 hours 5 minutes (range 11 minutes to 17 hours 47 minutes). In 146 (42%) patients the decision to transfer was primarily made by the emergency medicine clinician. A total of 251 (72%) patients were intubated. The documented critical incident rate was 15%. CONCLUSION: Trauma is the most common reason for transfer of the critically ill adult from the ED. A significant number of patients are transferred, however, with medical and surgical conditions and for non-clinical reasons. There continues to be problems with the quality of care that these patients receive. Emergency medicine clinicians must be actively involved in the development of regional critical care systems as a significant proportion of all critically ill adults transferred originate from the ED. PMID- 12748141 TI - Fasciotomy in crush injury resulting from prolonged pressure in an earthquake in Turkey. AB - OBJECTIVES: The authors report on the patients treated in a clinic who were injured in the earthquake that took place in north western part of Turkey in 1999 and was reported to be measured at 7.4 on the Richter scale. This catastrophe killed over 16 000 people while injuring more than 23 000 people. The type of housing was varied and entrapment occurred in single, two or more storey buildings. METHODS: 35 patients were admitted to the plastic surgery department after the earthquake. The hospital is about 400 km from the disaster site. Although all the transported victims had intravenous lines, few had adequate volume replacement. Sixteen had an urgent fasciotomy at the time of admission. The procedures were performed between 8 and 21 hours after extrication. After fasciotomy, all non-viable muscle content was removed but an attempt was made to retain as much viable muscle as possible. RESULTS: Amputation was required on four patients (25%). Fasciotomy incisions were closed with skin grafting in eight cases, and with primary closure in four cases. CONCLUSION: Prompt fasciotomy in earthquake victims will be both life saving and can prevent some of the severe and dangerous complications after crush syndrome. PMID- 12748140 TI - Crush syndrome patients after the Marmara earthquake. AB - BACKGROUND: To assess the treatment and outcome of patients with crush injury sustained in the Marmara earthquake. METHODS: Seven hundred eighty three patients were transferred to a university hospital and 25 of them were admitted to the intensive care unit. The medical records of 18 crush injury patients were retrospectively reviewed. RESULTS: The major associated injuries were in the lower extremities, upper extremities, and chest. Seven patients underwent fasciotomy and six patients had amputations. Twelve patients required mechanical ventilation. Adult respiratory distress syndrome developed in four patients. Oliguria occurred in eight patients. Hyperkalaemia was seen in six patients and four of them underwent emergency haemodialysis. One patient died because of hyperkalaemia on arrival to the intensive care unit. Renal failure was treated with haemodialysis or haemoperfusion in 13 patients. Five patients died because of multiple organ failure and two patients because of sepsis. CONCLUSION: Crush syndrome is a life threatening event. The authors believe that early transportation and immediate intensive care therapy would have improved the survival rate. PMID- 12748143 TI - Communication with general practitioners after accident and emergency attendance: computer generated letters are often deficient. AB - OBJECTIVES: Accident and emergency departments see large numbers of patients, and computerised administration systems are a useful tool for producing discharge communication. The purpose of this study was to determine the quality of such correspondence. METHODS: Retrospective review of 300 discharge letters and case notes. RESULTS: 29% of all computer generated discharge information was incomplete or misleading. Twenty five per cent of all correspondence was lacking or unacceptable overall. The principal reasons for substandard correspondence were inaccurate coding of diagnoses and procedures, and failure to include specific information relevant to patients' follow up. CONCLUSIONS: Computer generated discharge communication is often deficient. Staff using such systems should be made aware of the importance of accurate coding, and use added explanatory text to clarify diagnoses, management, and follow up as required. PMID- 12748142 TI - Barking up the wrong tree? A survey of dog bite wound management. AB - OBJECTIVES: Several trials have been published examining the role of antibiotics in dog bite wound management. A meta-analysis of these suggests that there is very little benefit to routine antibiotic prescription in these patients. All papers however incorporated rigorous wound care regimens involving large volume irrigation. METHODS: The authors undertook a telephone survey to investigate wound care and prescribing practice in bite wound management in accident and emergency and minor injury units in the Yorkshire region. RESULTS: Twenty one departments were contacted. Only 10% of departments routinely irrigate these wounds. Antibiotics are prescribed routinely in 71%. CONCLUSIONS: Management of dog bite wounds would not seem to be evidence based in most departments in this sample. PMID- 12748145 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Diamorphine or morphine for ischaemic cardiac chest pain. AB - A short cut review was carried out to establish whether morphine is better than diamorphine at allieviating chest pain after an acute myocardial infarction. Altogether 66 papers were found using the reported search, of which one presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of this best paper are tabulated. A clinical bottom line is stated. PMID- 12748144 TI - Statistical consideration for research. AB - The seventh paper in this series discusses the importance of statistical techniques in research. PMID- 12748147 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Glucagon in tricyclic overdose. AB - A short cut review was carried out to establish whether the addition of glucagon to standard treatments improves clinical outcome in patients who have taken an overdose of tricyclic antidepressants. Altogether 31 papers were found using the reported search, of which three presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 12748148 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Type of oral cortiosteroid in mild to moderate croup. AB - A short cut review was carried out to establish whether oral dexamethasone is better than oral prednisolone at improving outcome in children with mild to moderate croup. Altogether 139 papers were found using the reported search, of which none presented any evidence to answer the clinical question. It is concluded that there is no evidence available to answer this question. Further research is needed. PMID- 12748149 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Colourimetric CO(2) detector compared with capnography for confirming ET tube placement. AB - A short cut review was carried out to establish whether colourimetric carbon dioxide detectors are as reliable as capnometry at verifying tracheal placement of endotracheal tubes after emergency intubation. A total of 69 papers were found using the reported search, of which four presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 12748150 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Glucagon for the treatment of symptomatic beta blocker overdose. AB - A short cut review was carried out to establish whether the intravenous glucagon can support blood pressure in beta blocker overdose. A total of 51 papers were found using the reported search, of which six presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of these best papers are tabulated. A clinical bottom line is stated. PMID- 12748151 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Buscopan (hyoscine butylbromide) in abdominal colic. AB - A short cut review was carried out to establish whether buscopan (hyoscine butylbromide) is better than analgesics at controlling pain in abdominal colic. A total of 31 papers were found using the reported search, of which none presented any evidence to answer the clinical question. It is concluded that there is no evidence available to answer this question. Further research is needed. PMID- 12748152 TI - Observational study of prehospital delays in patients with chest pain. AB - OBJECTIVE: To define and measure patient reported prehospital delay in presentation to the emergency department with chest pain and identify simple strategies that may reduce this delay. The authors investigated the null hypothesis that the patients choice of service to call for acute medical help has no effect on the timing of thrombolysis. METHOD: A prospective observational study of prehospital times and events was undertaken on a target population of patients presenting with acute chest pain attributable to an acute coronary syndrome over a three month period. RESULTS: Patients who decided to call the ambulance service were compared with patients who contacted any other service. Most patients who contact non-ambulance services are seen by general practitioners. The prehospital system time for 121 patients who chose to call the ambulance service first was significantly shorter than for 96 patients who chose to call another service (median 57 min v 107 min; p<0.001). Of the 42 patients thrombolysed in the emergency department, those who chose to call the ambulance service had significantly shorter prehospital system times (number 21 v 21; median 44 v 69 min; p<0.001). Overall time from pain onset to initiation of thrombolysis was significantly longer in the group of patients who called a non ambulance service first (median 130 min v 248 min; p=0.005). CONCLUSIONS: Patient with acute ischaemic chest pain who call their general practice instead of the ambulance service are likely to have delayed thrombolysis. This is likely to result in increased mortality. The most beneficial current approach is for general practices to divert all patients with possible ischaemic chest pain onset within 12 hours direct to the ambulance service. PMID- 12748153 TI - Outcome of diabetic patients treated in the prehospital arena after a hypoglycaemic episode, and an exploration of treat and release protocols: a review of the literature. AB - OBJECTIVES: This review examines current treat and release protocols adopted by the ambulance service, and factors that may predispose patients to hypoglycaemia. METHODS: Online database searches and hand searches of journals led to 241 articles being found, of which eight were used for this article. RESULTS: Out of hospital treatment of hypoglycaemia is safe for most patients, but further studies are needed if positive improvements are to be made. DISCUSSION: There is potential for further research in this area and clarification is needed in the treat and release debate. A definitive set of protocols would be beneficial to the ambulance service. PMID- 12748154 TI - Emergency ambulances on the public highway linked with inconvenience and potential danger to road users. AB - OBJECTIVES: The main aim was to survey interactions between the public highway users and emergency ambulances using lights and sirens. The objectives were to identify negative and positive experiences, and to assess the frequency and consequences of these events. METHODS: Because of a poor response from a random postal pilot, a quota sample of 200 was adopted with a response of 65%. This provided data on demographics, details of previous interactions, and possible third party effects. Participant perception of psychological stress in negative interactions was also recorded. Possible links between the recorded interactions were assessed using the chi(2) tests of association. RESULTS: The passage of an emergency ambulance using lights and sirens caused the public to move from their chosen position in most cases (61%). Horns or sirens were used frequently (86%), but they were not always applied in unison with the warning lights. A significant association was found between the satisfactory handling of events by the ambulance crews and the use of audible warning devices (p<0.001). Twenty per cent of avoidance manoeuvres necessitated reversing, which was associated with a third party affect (p<0.005). Roughly one third of the participants found interactions stressful and felt that the events could have been avoided. Most public road users (91%) believed that they acted in a controlled manner. CONCLUSIONS: The findings suggest that most participants had interacted with the emergency ambulances in a positive manner, while a smaller but significant fraction of the public road users found the interactions difficult to handle. A third party effect was identified in avoidance manoeuvres. Further longitudal research with random sampling is recommended. PMID- 12748155 TI - The Scottish mountain rescue casualty study. AB - AIM: To describe injuries and illnesses in casualties rescued by Scottish mountain rescue teams during 1998 and 1999, with particular emphasis on major trauma. METHODS: Retrospective study. Information from mountain rescue reports, Scottish Trauma Audit Group database, and hospital case notes. RESULTS: Teams undertook 622 emergency callouts in the two years. A total of 333 casualties with injuries and illnesses rescued. There were 57 fatal incidents, 261 (78.4%) rescued with traumatic injuries, 12 (3.6%) suffering from major trauma, and 12 (3.6%) had spinal injuries. Half had lower limb injuries. Twenty six (7.7%) were suffering from non-traumatic medical problems. Forty six (13.8%) were suffering from cold or exhaustion. Fifty three casualties were dead when the rescue team arrived. Four died during or after rescue, one from hypothermia and three from trauma. All major trauma casualties were evacuated by helicopter. DISCUSSION: No previous similar studies identified. Significant numbers of seriously injured and ill casualties are being cared for by mountain rescue team casualty carers, many of whom are not healthcare professionals. The need for improved training, research, and equipment is discussed. CONCLUSION: Scottish MRTs are called upon to provide an advanced level of care for a significant number of casualties. There is a need for formalised opportunities for in hospital training, management protocols, and continuing research and audit-none of which currently exists. PMID- 12748156 TI - Do callers to NHS Direct follow the advice to attend an accident and emergency department? AB - OBJECTIVES: To provide an objective assessment on callers' compliance with NHS Direct advice to attend an accident and emergency (A&E) department. METHODS: A representative three week period in May 2000 was investigated. During this period there were no health scares, major health campaigns, or bank holidays that may have affected the call rate. NHS Direct callers who were advised to attend A&E were identified. Data from the four A&E departments for the same three week period and two additional days were searched and matched to NHS Direct data by surname, date of birth, and post code. This process created three groups: (1) callers triaged to A&E who attended, (2) callers triaged to A&E who did not attend, (3) callers with different triage outcome who attended A&E. The age, sex, relationship of caller, time of call, and distance to nearest A&E were compared for groups (1) and (2). RESULTS: Just less than two third of callers triaged to A&E attended with the same presenting complaint. There were no statistically significant differences between group (1) and (2) in terms of age, sex, relationship of caller, time of call, and distance to A&E. A small number of callers (2.4%) were identified as being given other advice and attending A&E for the same presenting complaint. This group took significantly longer to attend A&E than group (1) (chi(2) =139.01, df=7, p<0.001). CONCLUSIONS: Assessing levels of compliance is difficult. These findings suggest that NHS Direct may have comparatively high levels of compliance compared with other similar services. However, using the single triage outcome as the means of identifying the advice given may oversimplify the range of possible advice given. The delay in attending A&E for the group of callers who were given other advice may indicate they had tried other actions. Further larger studies are needed to assess the appropriateness of referrals through investigation of clinical outcomes. PMID- 12748158 TI - The point of the needle. Occult pneumothorax: a review. AB - The case of a patient with an unusual medical condition and an occult pneumothorax is presented. The evidence for management of occult pneumothorax particularly in patients with underlying lung disease is reviewed and solutions to the acute clinical problems that may arise are suggested. PMID- 12748157 TI - NHS Direct: consistency of triage outcomes. AB - OBJECTIVES: To examine the consistency of triage outcomes by nurses using four types of computerised decision support software in NHS Direct. METHODS: 119 scenarios were constructed based on calls to ambulance services that had been assigned the lowest priority category by the emergency medical dispatch systems in use. These scenarios were presented to nurses working in four NHS Direct call centres using different computerised decision support software, including the NHS Clinical Assessment System. RESULTS: The overall level of agreement between the nurses using the four systems was "fair" rather than "moderate" or "good" (kappa=0.375, 95% CI: 0.34 to 0.41). For example, the proportion of calls triaged to accident and emergency departments varied from 22% (26 of 119) to 44% (53 of 119). Between 21% (25 of 119) and 31% (37 of 119) of these low priority ambulance calls were triaged back to the 999 ambulance service. No system had both high sensitivity and specificity for referral to accident and emergency services. CONCLUSIONS: There were large differences in outcome between nurses using different software systems to triage the same calls. If the variation is primarily attributable to the software then standardising on a single system will obviously eliminate this. As the calls were originally made to ambulance services and given the lowest priority, this study also suggests that if, in the future, ambulance services pass such calls to NHS Direct then at least a fifth of these may be passed back unless greater sensitivity in the selection of calls can be achieved. PMID- 12748159 TI - Caesarean delivery during maternal cardiopulmonary resuscitation for status asthmaticus. AB - A patient who sustained a recurrent cardiopulmonary resuscitation due to status asthmaticus during one pregnancy followed by a birth of an apparently normal infant is described. Promptly performed caesarean delivery might have saved the mother and her infant. Cardiopulmonary resuscitation is less effective in a near term pregnant woman. PMID- 12748160 TI - Three women in a car. PMID- 12748162 TI - Stridor and Down's syndrome. PMID- 12748161 TI - Temperate pyomyositis in an injecting drug misuser. A difficult diagnosis in a difficult patient. AB - The medical care of injecting drug misusers presents many challenges. Though they can be awkward and unreliable they are at risk of serious medical conditions not often seen in the general population. This case report illustrates some of the difficulties in the diagnosis and treatment of a patient with pyomyositis associated with heroin injection. PMID- 12748163 TI - Public health warning: Pancake Day! PMID- 12748164 TI - What is the patient's best telephone number? PMID- 12748165 TI - Cyclizine. PMID- 12748166 TI - Co-trimoxazole induced acute thrombocytopenic purpura. PMID- 12748167 TI - Successful resuscitation of a patient with electrical storm. AB - A 41 year old woman with type 2 diabetes, hypertension, and hyperlipidaemia but no known heart disease received 130 DC shocks for repeated cardiac arrests due to ventricular tachyarrhythmias over 48 hours. She was stabilised by intravenous amiodarone and had a defibrillator implanted. Serial ECGs did not change, but raised troponin I confirmed myocardial infarction as the underlying cause. Electrical storm is an uncommon and dramatic but usually treatable syndrome of recurrent ventricular arrhythmias. Frequent precipitants of electrical storm include recent worsening heart failure, hypokalaemia, hypomagnesaemia and myocardial ischaemia. Amiodarone is the antiarrhythmic agent of choice and implantable cardioverter defibrillator improves long term outcome. PMID- 12748168 TI - HU protein of Escherichia coli has a role in the repair of closely opposed lesions in DNA. AB - Closely opposed lesions form a unique class of DNA damage that is generated by ionizing radiation. Improper repair of closely opposed lesions could lead to the formation of double strand breaks that can result in increased lethality and mutagenesis. In vitro processing of closely opposed lesions was studied using double-stranded DNA containing a nick in close proximity opposite to a dihydrouracil. In this study we showed that HU protein, an Escherichia coli DNA binding protein, has a role in the repair of closely opposed lesions. The repair of dihydrouracil is initiated by E. coli endonuclease III and processed via the base excision repair pathway. HU protein was shown to inhibit the rate of removal of dihydrouracil by endonuclease III only when the DNA substrate contained a nick in close proximity opposite to the dihydrouracil. In contrast, HU protein did not inhibit the subsequent steps of the base excision repair pathway, namely the DNA synthesis and ligation reactions catalyzed by E. coli DNA polymerase and E. coli DNA ligase, respectively. The nick-dependent selective inhibition of endonuclease III activity by HU protein suggests that HU could play a role in reducing the formation of double strand breaks in E. coli. PMID- 12748169 TI - Tumor necrosis factor-induced nuclear factor kappaB activation is impaired in focal adhesion kinase-deficient fibroblasts. AB - Focal adhesion kinase (FAK) is widely involved in important cellular functions such as proliferation, migration, and survival, although its roles in immune and inflammatory responses have yet to be explored. We demonstrate a critical role for FAK in the tumor necrosis factor (TNF)-induced activation of nuclear factor (NF)-kappaB, using FAK-deficient (FAK-/-) embryonic fibroblasts. Interestingly, TNF-induced interleukin (IL)-6 production was nearly abolished in FAK-/- fibroblasts, whereas a normal level of production was obtained in FAK+/- or FAK+/+ fibroblasts. FAK deficiency did not affect the three types of mitogen activated protein kinases, ERK, JNK, and p38. Similarly, TNF-induced activation of activator protein 1 or NF-IL-6 was not impaired in FAK-/- cells. Of note, TNF induced NF-kappaB DNA binding activity and activation of IkappaB kinases (IKKs) were markedly impaired in FAK-/- cells, whereas the expression of TNF receptor I or other signaling molecules such as receptor-interacting protein (RIP), tumor necrosis factor receptor-associated factor 2 (TRAF2), IKKalpha, IKKbeta, and IKKgamma was unchanged. Also, TNF-induced association of FAK with RIP and subsequent association of RIP with TRAF2 were not observed, resulting in a failure of RIP to recruit the IKK complex in FAK-/- cells. The reintroduction of wild type FAK into FAK-/- cells restored the interaction of RIP with TRAF2 and the IKK complex and allowed recovery of NF-kappaB activation and subsequent IL-6 production. Thus, we propose a novel role for FAK in the NF-kappaB activation pathway leading to the production of cytokines. PMID- 12748170 TI - Evidence for the role of a peroxidase compound I-type intermediate in the oxidation of glutathione, NADH, ascorbate, and dichlorofluorescin by cytochrome c/H2O2. Implications for oxidative stress during apoptosis. AB - The release of cytochrome c from mitochondria is a crucial step in apoptosis, resulting in the activation of the caspase proteases. A further consequence of cytochrome c release is the enhanced mitochondrial production of superoxide radicals (O2.), which are converted to hydrogen peroxide by manganese-superoxide dismutase. Recently, we showed that cytochrome c is a potent catalyst of 2',7' dichlorofluorescin oxidation to the fluorescent 2',7'-dichlorofluorescein by these species, leading to the conclusion that 2',7'-dichlorofluorescein fluorescence is a reflection of cytosolic cytochrome c concentration rather than "reactive oxygen species" levels (Burkitt, M. J., and Wardman, P. (2001) Biochem. Biophys. Res. Commun. 282, 329-333). The oxidant generated from cytochrome c has so far not been identified. Several authors have suggested that the hydroxyl radical (*OH) is generated, but others have discussed the possibility of a peroxidase compound I. By examining the effects of various antioxidants (glutathione, ascorbate, and NADH) and "hydroxyl radical scavengers" (ethanol and mannitol) on the rate of 2',7'-dichlorofluorescin oxidation by cytochrome c, together with complementary EPR spin-trapping studies, we demonstrate that the hydroxyl radical is not generated. Instead, our findings suggest the formation of a peroxidase compound I-type intermediate, in which one oxidizing equivalent is present as an oxoferryl heme species and the other as the protein tyrosyl radical previously identified (Barr, D. P., Gunther, M. R., Deterding, L. J., Tomer, K. B., and Mason, R. P. (1996) J. Biol. Chem. 271, 15498-15503). Competition studies involving spin traps indicated that the oxoferryl heme component is the active oxidant. These findings provide an improved understanding of the physicochemical basis of the redox changes that occur during apoptosis. PMID- 12748171 TI - An aminopeptidase, ARTS-1, is required for interleukin-6 receptor shedding. AB - Aminopeptidase regulator of TNFR1 shedding (ARTS-1) binds to the type I tumor necrosis factor receptor (TNFR1) and promotes receptor shedding. Because hydroxamic acid-based metalloprotease inhibitors prevent shedding of both TNFR1 and the interleukin-6 receptor (IL-6Ralpha), we hypothesized that ARTS-1 might also regulate shedding of IL-6Ralpha, a member of the type I cytokine receptor superfamily that is structurally different from TNFR1. Reciprocal co immunoprecipitation experiments identified that membrane-associated ARTS-1 directly binds to a 55-kDa IL-6Ralpha, a size consistent with soluble IL-6Ralpha generated by ectodomain cleavage of the membrane-bound receptor. Furthermore, ARTS-1 promoted IL-6Ralpha shedding, as demonstrated by a direct correlation between increased membrane-associated ARTS-1 protein, increased IL-6Ralpha shedding, and decreased membrane-associated IL-6Ralpha in cell lines overexpressing ARTS-1. The absence of basal IL-6Ralpha shedding from arts-1 knock out cells identified that ARTS-1 was required for constitutive IL-6Ralpha shedding. Furthermore, the mechanism of constitutive IL-6Ralpha shedding requires ARTS-1 catalytic activity. Thus, ARTS-1 promotes the shedding of two cytokine receptor superfamilies, the type I cytokine receptor superfamily (IL-6Ralpha) and the TNF receptor superfamily (TNFR1). We propose that ARTS-1 is a multifunctional aminopeptidase that may modulate inflammatory events by promoting IL-6Ralpha and TNFR1 shedding. PMID- 12748172 TI - Identification and characterization of a nuclear interacting partner of anaplastic lymphoma kinase (NIPA). AB - Anaplastic large-cell lymphoma is a subtype of non-Hodgkin lymphomas characterized by the expression of CD30. More than half of these lymphomas carry a chromosomal translocation t(2;5) leading to expression of the oncogenic tyrosine kinase nucleophosmin-anaplastic lymphoma kinase (NPM-ALK). NPM-ALK is capable of transforming fibroblasts and lymphocytes in vitro and of causing lymphomas in mice. Previously, we and others demonstrated phospholipase C-gamma and phosphatidylinositol 3-kinase as crucial downstream signaling mediators of NPM-ALK-induced oncogenicity. In this study, we used an ALK fusion protein as bait in a yeast two-hybrid screen identifying NIPA (nuclear interacting partner of ALK) as a novel downstream target of NPM-ALK. NIPA encodes a 60-kDa protein that is expressed in a broad range of human tissues and contains a classical nuclear translocation signal in its C terminus, which directs its nuclear localization. NIPA interacts with NPM-ALK and other ALK fusions in a tyrosine kinase-dependent manner and is phosphorylated in NPM-ALK-expressing cells on tyrosine and serine residues with serine 354 as a major phosphorylation site. Overexpression of NIPA in Ba/F3 cells was able to protect from apoptosis induced by IL-3 withdrawal. Mutations of the nuclear translocation signal or the Ser-354 phosphorylation site impaired the antiapoptotic function of NIPA. In NPM-ALK transformed Ba/F3 cells, apoptosis triggered by wortmannin treatment was enhanced by overexpression of putative dominant-negative NIPA mutants. These results implicate an antiapoptotic role for NIPA in NPM-ALK-mediated signaling events. PMID- 12748173 TI - Transcriptional regulation of interleukin (IL)-8 by bradykinin in human airway smooth muscle cells involves prostanoid-dependent activation of AP-1 and nuclear factor (NF)-IL-6 and prostanoid-independent activation of NF-kappaB. AB - Bradykinin (BK) is a potent neutrophil chemotractant, proinflammatory mediator, and angiogenic factor, which acts through G protein-coupled receptors (GPCRs). Here we studied the mechanisms involved in IL-8 generation by BK in human airway smooth muscle cells focusing on the transcription factors involved and role of endogenous prostanoids in transcription factor activation. Transfection experiments with wild-type IL-8 promoter constructs or constructs with NF-kappaB, AP-1, and NF-IL-6 binding site mutations suggested that all three transcription factors were necessary for optimal IL-8 expression. BK increased NF-kappaB, AP-1, and NF-IL-6 binding to the IL-8 promoter by electrophoretic mobility shift assay. NF-kappaB, the most important transcription factor in the current study, was translocated to the nucleus after BK stimulation. Indomethacin, a cyclooxygenase inhibitor, partially inhibited IL-8 release and the promoter binding of AP-1 and NF-IL-6, but not NF-kappaB. Furthermore, exogenous prostaglandin E2 stimulated AP 1 and NF-IL-6 binding to the IL-8 promoter. The anti-inflammatory glucocorticoid dexamethasone inhibited NF-kappaB translocation and the promoter binding of NF kappaB, AP-1, and NF-IL-6. These results are the first to delineate the transcription factors involved in BK induced IL-8 release. Transcriptional activation of the IL-8 promoter by BK involves the prostanoid-independent activation of NF-kappaB, and prostanoid-dependent activation of AP-1 and NF-IL-6 plays a key role in augmenting the response. Endogenous prostanoid generation in response to GPCR ligands such as BK may be an important mechanism whereby GPCRs signal to the nucleus to maximize the transcription of inflammatory response genes. PMID- 12748174 TI - Membrane-anchoring and charge effects in the interaction of myelin basic protein with lipid bilayers studied by site-directed spin labeling. AB - Myelin basic protein (MBP) maintains the compaction of the myelin sheath in the central nervous system by anchoring the cytoplasmic face of the two apposing bilayers and may also play a role in signal transduction. Site-directed spin labeling was done at eight matching sites in each of two recombinant murine MBPs, qC1 (charge +19) and qC8 charge (+13), which, respectively, emulate the native form of the protein (C1) and a post-translationally modified form (C8) that is increased in multiple sclerosis. When interacting with large unilamellar vesicles, most spin-labeled sites in qC8 were more mobile than those in qC1. Depth measurement via continuous wave power saturation indicated that the N terminal and C-terminal sites in qC1 were located below the plane of the phospholipid headgroups. In qC8, the C-terminal domain dissociated from the membrane, suggesting a means by which the exposure of natural C8 to cytosolic enzymes and ligands might increase in vivo in multiple sclerosis. The importance of two Phe-Phe pairs in MBP to its interactions with lipids was investigated by separately mutating each pair to Ala-Ala. The mobility at F42A/F43A and especially F86A/F87A increased significantly. Depth measurements and helical wheel analysis indicated that the Phe-86/Phe-87 region could form a surface seeking amphipathic alpha-helix. PMID- 12748175 TI - Is the Trypanosoma brucei REL1 RNA ligase specific for U-deletion RNA editing, and is the REL2 RNA ligase specific for U-insertion editing? AB - It was shown previously that the REL1 mitochondrial RNA ligase in Trypanosoma brucei was a vital gene and disruption affected RNA editing in vivo, whereas the REL2 RNA ligase gene could be down-regulated with no effect on cell growth or on RNA editing. We performed down-regulation of REL1 in procyclic T. brucei (midgut insect forms) by RNA interference and found a 40-50% inhibition of Cyb editing, which has only U-insertions, as well as a similar inhibition of ND7 editing, which has both U-insertions and U-deletions. In addition, both U-insertion and U deletion in vitro pre-cleaved editing were inhibited to similar extents. We also found little if any effect of REL1 down-regulation on the sedimentation coefficient or abundance of the RNA ligase-containing L-complex (Aphasizhev, R., Aphasizheva, I., Nelson, R. E., Gao, G., Simpson, A. M., Kang, X., Falick, A. M., Sbicego, S., and Simpson, L. (2003) EMBO J. 22, 913-924), suggesting that the inhibition of both insertion and deletion editing was not due to a disruption of the L-complex. Together with the evidence that down-regulation of REL2 has no effect on cell growth or on RNA editing in vivo or in vitro, these data suggest that the REL1 RNA ligase may be active in vivo in both U-insertion and U-deletion editing. The in vivo biological role of REL2 remains obscure. PMID- 12748176 TI - Identification and characterization of heme-interacting proteins in the malaria parasite, Plasmodium falciparum. AB - The degradation of hemoglobin by the malaria parasite, Plasmodium falciparum, produces free ferriprotoporphyrin IX (FP) as a toxic by-product. In the presence of FP-binding drugs such as chloroquine, FP detoxification is inhibited, and the build-up of free FP is thought to be a key mechanism in parasite killing. In an effort to identify parasite proteins that might interact preferentially with FP, we have used a mass spectrometry approach. Proteins that bind to FP immobilized on agarose include P. falciparum glyceraldehyde-3-phosphate dehydrogenase (PfGAPDH), P. falciparum glutathione reductase (PfGR), and P. falciparum protein disulfide isomerase. To examine the potential consequences of FP binding, we have examined the ability of FP to inhibit the activities of GAPDH and GR from P. falciparum and other sources. FP inhibits the enzymic activity of PfGAPDH with a Ki value of 0.2 microm, whereas red blood cell GAPDH is much less sensitive. By contrast, PfGR is more resistant to FP inhibition (Ki > 25 microm) than its human counterpart. We also examined the ability of FP to inhibit the activities of the additional antioxidant enzymes, P. falciparum thioredoxin reductase, which exhibits a Ki value of 1 microm, and P. falciparum glutaredoxin, which shows more moderate sensitivity to FP. The exquisite sensitivity of PfGAPDH to FP may indicate that the glycolytic pathway of the parasite is particularly susceptible to modulation by FP stress. Inhibition of this pathway may drive flux through the pentose phosphate pathway ensuring sufficient production of reducing equivalents to counteract the oxidative stress induced by FP build-up. PMID- 12748177 TI - Valproate induces replication-independent active DNA demethylation. AB - In this report, we demonstrate that valproic acid (VPA), a drug that has been used for decades in the treatment of epilepsy and as a mood stabilizer, triggers replication-independent active demethylation of DNA. Thus, this drug can potentially reverse DNA methylation patterns and erase stable methylation imprints on DNA in non-dividing cells. Recent discoveries support a role for VPA in the regulation of methylated genes; however, the mechanism has been unclear because it is difficult to dissociate active demethylation from the absence of DNA methylation during DNA synthesis. We therefore took advantage of an assay that measures active DNA demethylation independently from other DNA methylation and DNA replication activities in human embryonal kidney 293 cells. We show that VPA induces histone acetylation, DNA demethylation, and expression of an ectopically methylated CMV-GFP plasmid in a dose-dependent manner. In contrast, valpromide, an analogue of VPA that does not induce histone acetylation, does not induce demethylation or expression of CMV-GFP. Furthermore, we illustrate that methylated DNA-binding protein 2/DNA demethylase (MBD2/dMTase) participates in this reaction since antisense knockdown of MBD2/dMTase attenuates VPA-induced demethylation. Taken together, our data support a new mechanism of action for VPA as enhancing intracellular demethylase activity through its effects on histone acetylation and raises the possibility that DNA methylation is reversible independent of DNA replication by commonly prescribed drugs. PMID- 12748178 TI - Subunit structures and stoichiometries of human DNA fragmentation factor proteins before and after induction of apoptosis. AB - DNA fragmentation factor (DFF) is one of the major endonucleases responsible for internucleosomal DNA cleavage during apoptosis. Understanding the regulatory checkpoints involved in safeguarding non-apoptotic cells against accidental activation of this nuclease is as important as elucidating its activation mechanisms during apoptosis. Here we address these issues by determining DFF native subunit structures and stoichiometries in human cells before and after induction of apoptosis using the technique of native pore-exclusion limit electrophoresis in combination with Western analyses. For comparison, we employed similar techniques with recombinant proteins in conjunction with atomic force microscopy. Before induction of apoptosis, the expression of DFF subunits varied widely among the cell types studied, and the chaperone/inhibitor subunits DFF45 and DFF35 unexpectedly existed primarily as monomers in vast excess of the latent nuclease subunit, DFF40, which was stoichiometrically associated with DFF45 to form heterodimers. DFF35 was exclusively cytoplasmic as a monomer. Nuclease activation upon caspase-3 cleavage of DFF45/DFF35 was accompanied by DFF40 homo oligomer formation, with a tetramer being the smallest unit. Interestingly, intact DFF45 can inhibit nuclease activity by associating with these homo oligomers without mediating their disassembly. We conclude that DFF nuclease is regulated by multiple pre- and post-activation fail-safe steps. PMID- 12748179 TI - The human caspase-8 promoter sustains basal activity through SP1 and ETS-like transcription factors and can be up-regulated by a p53-dependent mechanism. AB - Caspase-8, also known as MACH/FLICE/Mch5, is the most upstream-located cysteine aspartyl-protease (caspase) in a caspase cascade involved in apoptosis triggered by members of the tumor necrosis factor receptor superfamily or other stimuli such as chemotherapeutic agents. Regulation of caspase-8 expression on a post translational level has been studied in detail, whereas only little information is available on its control by gene transcription. We identified and cloned the human caspase-8 promoter, determined the transcriptional start site of the caspase-8 gene, and examined the regulatory mechanisms of the promoter with respect to its basal activity as well as to its inducibility upon apoptotic stimuli in human hepatoma cells. We identified two minimal sequences essential for basal transcription of caspase-8 and demonstrate that a single SP1 and an ETS like binding motif mediate this effect. We further show that the caspase-8 promoter is inducible and demonstrate that adenoviral infection increases caspase 8 mRNA levels. However, the increase in caspase-8 gene transcription after adenoviral infection absolutely depends on the p53 status of the hepatoma cell line, implying that caspase-8 is a target gene of p53. We show that delivery of exogenous p53 alone is sufficient to induce the caspase-8 promoter even in p53 deficient Hep3B hepatoma cells. Subsequent promoter deletion analysis in combination with luciferase reporter assays identified a p53-responsive element downstream of the transcriptional start site. We demonstrate that this p53 responsive sequence overlaps with the ETS-like binding site and suggest that an additional p53-inducible, yet unknown factor interacts with this region of the caspase-8 promoter. In summary, our study contributes to the understanding of the transcriptional regulation of the caspase-8 gene by basal (SP1- and ETS dependent) and inducible (p53-dependent) mechanisms. PMID- 12748180 TI - Systemic signaling in tomato plants for defense against herbivores. Isolation and characterization of three novel defense-signaling glycopeptide hormones coded in a single precursor gene. AB - An 18-amino acid peptide in tomato leaves called systemin is a primary signal released at wound sites in response to herbivory that systemically signals the activation of defense genes throughout the plants. We report here the isolation of three hydroxyproline-rich glycopeptides from tomato leaves, of 20, 18, and 15 amino acids in length, that signal the activation of defense genes, similar to the activity of the systemin peptide. The three new peptides cause an alkalinization of suspension-cultured cells and induce the synthesis of defensive proteinase inhibitor proteins when supplied at fmol levels to young tomato plants through their cut stems. This suggests that they are part of the wound signaling of tomato plants that activates defense against herbivores and pathogens. Isolation of cDNAs coding for the tomato peptides revealed that they are all derived from the same pre-proprotein precursor that is systemically wound inducible. The peptides are considered members of the functionally characterized systemin family of defense signals from plants that are synthesized both in wounded leaves and in distal, unwounded leaves in response to herbivory or other mechanical wounding. The precursor deduced from the cDNA exhibits a leader sequence, indicating that it is synthesized through the secretory pathway, where it is hydroxylated and glycosylated. The amino acid sequence of the precursor exhibited weak identity to the precursor of two hydroxyproline-rich defense signals recently found in tobacco, suggesting that the two pre-protein precursors have evolved from a common ancestral protein. The identification of hydroxyproline-rich glycoprotein systemins in tomato indicates that the initiation of wound signaling is more complex than previously thought and appears to involve multiple peptide signals. PMID- 12748181 TI - The different large subunit isoforms of Arabidopsis thaliana ADP-glucose pyrophosphorylase confer distinct kinetic and regulatory properties to the heterotetrameric enzyme. AB - ADP-glucose pyrophosphorylase catalyzes the first and limiting step in starch biosynthesis and is allosterically regulated by the levels of 3-phosphoglycerate and phosphate in plants. ADP-glucose pyrophosphorylases from plants are heterotetramers composed of two types of subunits (small and large). In this study, the six Arabidopsis thaliana genes coding for ADP-glucose pyrophosphorylase isoforms (two small and four large subunits) have been cloned and expressed in an Escherichia coli mutant deficient in ADP-glucose pyrophosphorylase activity. The co-expression of the small subunit APS1 with the different Arabidopsis large subunits (APL1, APL2, APL3, and APL4) resulted in heterotetramers with different regulatory and kinetic properties. Heterotetramers composed of APS1 and APL1 showed the highest sensitivity to the allosteric effectors as well as the highest apparent affinity for the substrates (glucose-1 phosphate and ATP), whereas heterotetramers formed by APS1 and APL2 showed the lower response to allosteric effectors and the lower affinity for the substrates. No activity was detected for the second gene coding for a small subunit isoform (APS2) annotated in the Arabidopsis genome. This lack of activity is possibly due to the absence of essential amino acids involved in catalysis and/or in the binding of glucose-1-phosphate and 3-phosphoglycerate. Kinetic and regulatory properties of the different heterotetramers, together with sequence analysis has allowed us to make a distinction between sink and source enzymes, because the combination of different large subunits would provide a high plasticity to ADP glucose pyrophosphorylase activity and regulation. This is the first experimental data concerning the role that all the ADP-glucose pyrophosphorylase isoforms play in a single plant species. This phenomenon could have an important role in vivo, because different large subunits would confer distinct regulatory properties to ADP-glucose pyrophosphorylase according to the necessities for starch synthesis in a given tissue. PMID- 12748182 TI - Factors controlling the uptake of yeast copper/zinc superoxide dismutase into mitochondria. AB - We have previously shown that a fraction of yeast copper/zinc-superoxide dismutase (SOD1) and its copper chaperone CCS localize to the intermembrane space of mitochondria. In the present study, we have focused on the mechanism by which SOD1 is partitioned between cytosolic and mitochondrial pools. Using in vitro mitochondrial import assays, we show that only a very immature form of the SOD1 polypeptide that is apo for both copper and zinc can efficiently enter the mitochondria. Moreover, a conserved disulfide in SOD1 that is essential for activity must be reduced to facilitate mitochondrial uptake of SOD1. Once inside the mitochondria, SOD1 is converted to an active holo enzyme through the same post-translational modifications seen with cytosolic SOD1. The presence of high levels of CCS in the mitochondrial intermembrane space results in enhanced mitochondrial accumulation of SOD1, and this apparently involves CCS-mediated retention of SOD1 within mitochondria. This retention of SOD1 is not dependent on copper loading of the enzyme but does require protein-protein interactions at the heterodimerization interface of SOD1 and CCS as well as conserved cysteine residues in both molecules. A model for how CCS-mediated post-translational modification of SOD1 controls its partitioning between the mitochondria and cytosol will be presented. PMID- 12748183 TI - Intramolecular higher order packing of parallel quadruplexes comprising a G:G:G:G tetrad and a G(:A):G(:A):G(:A):G heptad of GGA triplet repeat DNA. AB - GGA triplet repeats are widely dispersed throughout eukaryotic genomes and are frequently located within biologically important regions such as gene regulatory regions and recombination hot spot sites. We determined the structure of d(GGA)4 (12-mer) under physiological conditions and founded the formation of an intramolecular parallel quadruplex for the first time. Later, a similar architecture to that of the intramolecular parallel quadruplex was found for a telomere DNA in the crystalline state. Here, we have determined the structure of d(GGA)8 (24-mer) under physiological conditions. Two intramolecular parallel quadruplexes comprising a G:G:G:G tetrad and a G(:A):G(:A):G(:A):G heptad are formed in d(GGA)8. These quadruplexes are packed in a tail-to-tail manner. This is the first demonstration of the intramolecular higher order packing of quadruplexes at atomic resolution. K+ ions, but not Na+ ones, are critically required for the formation of this unique structure. The elucidated structure suggests the mechanisms underlying the biological events related to the GGA triplet repeat. Furthermore, in the light of the structure, the mode of the higher order packing of the telomere DNA is discussed. PMID- 12748184 TI - Hyaluronan-mediated CD44 interaction with RhoGEF and Rho kinase promotes Grb2 associated binder-1 phosphorylation and phosphatidylinositol 3-kinase signaling leading to cytokine (macrophage-colony stimulating factor) production and breast tumor progression. AB - In this study we have examined CD44 (a hyaluronan (HA) receptor) interaction with a RhoA-specific guanine nucleotide exchange factor (p115RhoGEF) in human metastatic breast tumor cells (MDA-MB-231 cell line). Immunoprecipitation and immunoblot analyses indicate that both CD44 and p115RhoGEF are expressed in MDA MB-231 cells and that these two proteins are physically associated as a complex in vivo. The binding of HA to MDA-MB-231 cells stimulates p115RhoGEF-mediated RhoA signaling and Rho kinase (ROK) activity, which, in turn, increases serine/threonine phosphorylation of the adaptor protein, Gab-1 (Grb2-associated binder-1). Phosphorylated Gab-1 promotes PI 3-kinase recruitment to CD44v3. Subsequently, PI 3-kinase is activated (in particular, alpha, beta, gamma forms but not the delta form of the p110 catalytic subunit), AKT signaling occurs, the cytokine (macrophage-colony stimulating factor (M-CSF)) is produced, and tumor cell-specific phenotypes (e.g. tumor cell growth, survival and invasion) are up regulated. Our results also demonstrate that HA/CD44-mediated oncogenic events (e.g. AKT activation, M-CSF production and breast tumor cell-specific phenotypes) can be effectively blocked by a PI 3-kinase inhibitor (LY294002). Finally, we have found that overexpression of a dominant-negative form of ROK (by transfection of MBA-MD-231 cells with the Rho-binding domain cDNA of ROK) not only inhibits HA/CD44-mediated RhoA-ROK activation and Gab-1 phosphorylation but also down-regulates oncogenic signaling events (e.g. Gab-1.PI 3-kinase-CD44v3 association, PI 3-kinase-mediated AKT activation, and M-CSF production) and tumor cell behaviors (e.g. cell growth, survival, and invasion). Taken together, these findings strongly suggest that CD44 interaction with p115RhoGEF and ROK plays a pivotal role in promoting Gab-1 phosphorylation leading to Gab-1.PI 3-kinase membrane localization, AKT signaling, and cytokine (M-CSF) production during HA mediated breast cancer progression. PMID- 12748185 TI - Muc4/sialomucin complex, the intramembrane ErbB2 ligand, translocates ErbB2 to the apical surface in polarized epithelial cells. AB - Muc4/Sialomucin complex (SMC) acts as an intramembrane ligand for the receptor tyrosine kinase ErbB2, inducing a limited phosphorylation of the receptor. Because Muc4/SMC is found at the apical surfaces of polarized epithelial cells and ErbB2 is often basolateral, the question arises as to whether these components become associated in polarized cells. To address this question, we examined the localization of these proteins in polarized human colon carcinoma CACO-2 cells. Dual color immunofluorescence analysis by confocal microscopy demonstrated the basolateral localization of the ErbB2 in these cells; it is primarily co-localized with E-cadherin at adherens junctions. Expression of apical Muc4/SMC in these cells by transient transfection results in the localization of the ErbB2 at the apical surface. Two-color confocal microscopy indicated that ErbB2 is colocalized with Muc4/SMC in the transfected cells but not in untransfected cells in the same culture. The change of localization of ErbB2 was confirmed by cell surface biotinylation of apical and basolateral proteins, followed by streptavidin precipitation and the subsequent detection of ErbB2 by immunoblotting. In contrast, Na+/K+-ATPase maintains its basolateral localization in Muc4/SMC-transfected cells, indicating that the translocation of ErbB2 is not the result of depolarization of the cells. A potential physiological role for the apical localization of ErbB2 is indicated by the fact that ErbB2 phosphorylated at tyrosine 1248 is found predominantly in Muc4/SMC-transfected cells, but not in untransfected cells, and is co-localized with the apical Muc4/SMC. The ability of Muc4/SMC to alter the localization of ErbB2, particularly a phosphorylated form of it, in epithelial cells, suggests that it has an important role in regulating ErbB2 signaling. PMID- 12748186 TI - Deficient regulation of DNA double-strand break repair in Fanconi anemia fibroblasts. AB - Fibroblasts from patients with Fanconi anemia (FA) display genomic instability, hypersensitivity to DNA cross-linking agents, and deficient DNA end joining. Fibroblasts from two FA patients of unidentified complementation group also had significantly increased cellular homologous recombination (HR) activity. Results described herein show that HR activity levels in patient-derived FA fibroblasts of groups A, C, and G were 10-fold greater than those seen in normal fibroblasts. In contrast, HR activity in group D2 fibroblasts was identical to that in normal cells. Western blot analysis revealed that the RAD51 protein was elevated 10-fold above normal levels in group A, C, and G fibroblasts, but was not altered in group D2 fibroblasts. HR activity levels in these former cells could be restored to near-normal levels by electroporation with anti-RAD51 antibody, whereas similar treatment of normal and complementation group D2 fibroblasts had no effect. These findings are consistent with a model in which FA proteins function to coordinate DNA double-strand break repair activity by regulating both recombinational and non-recombinational DNA repair. Interestingly, whereas positive regulation of DNA end joining requires the combined presence of all FA proteins thus far tested, suppression of HR, which is minimally dependent on the FANCA, FANCC, and FANCG proteins, does not require FANCD2. PMID- 12748187 TI - Identification of a novel Kruppel-associated box domain protein, Krim-1, that interacts with c-Myc and inhibits its oncogenic activity. AB - We have used the Ras recruitment system to screen for proteins that interact with the N-terminally located transactivation domain of c-Myc. The Ras recruitment system is based on the activation of the mitogenic RAS signaling pathway in yeast by the mammalian GTPase Ha-Ras. This screen led to the identification of two novel nuclear proteins termed Krim-1A and Krim-1B that both contain an N-terminal KRAB box domain and 12 or 9 Kruppel C2H2 type zinc fingers at the C terminus, respectively. We found that sequences covering the Myc box II homology region are essential for the interaction with the Krim-1 proteins and that the second N terminal zinc finger of Krim-1 is essential for Myc binding. Both Krim-1A and -B genes appear to be expressed ubiquitously with highest levels in spleen and lymph nodes. In particular, Krim-1B and, to a lesser extent, Krim-1A are able to decrease E-box-dependent transcriptional transactivation by c-Myc-Max complexes and also the ability of Myc to malignantly transform primary rat embryo fibroblasts, which is consistent with the functional repressive properties of their KRAB domains. The transcriptional corepressor Tif-1beta is a binding partner for Krim-1 and stabilizes the protein. Our findings suggest that Myc mediated functions can be negatively regulated by Krim-1, potentially in a complex with Tif-1beta. PMID- 12748188 TI - RING finger protein AO7 supports NF-kappaB-mediated transcription by interacting with the transactivation domain of the p65 subunit. AB - In this study, a novel interactor of the p65 subunit (RelA) of NF-kappaB has been explored by performing yeast two-hybrid screen using the transactivation domain (TAD) of p65 located in the C terminus as bait. We have isolated a RING finger motif-containing protein, AO7, previously identified as an interacting protein with a ubiquitin-conjugating enzyme, Ubc5B. We confirmed the protein-protein interaction between p65 and AO7 in vitro and in vivo and found that the C terminal region of AO7 is responsible for the interaction with p65 TAD. AO7 was predominantly localized in the nucleus and activated the NF-kappaB-dependent gene expression upon stimulation with IL-1beta or TNF or overexpression of NF-kappaB inducing kinase. We found that both the RING finger and the C-terminal regions of AO7 were necessary for the transcriptional activation. When cotransfected with plasmids expressing Gal4-p65 fusion proteins containing various functional domains of p65, we found that p65 TAD was essential for the transcriptional activation mediated by AO7. Furthermore, the p65-mediated transactivation was suppressed by a ubiquitination-defective AO7 mutant in which the essential Cys residue within the RING finger motif was substituted by Ser. These data suggest that AO7 interacts with the p65 TAD and modulates its transcriptional activity. PMID- 12748189 TI - Characterization of the DNA damage-inducible helicase DinG from Escherichia coli. AB - The dinG promoter was first isolated in a genetic screen scoring for damage inducible loci in Escherichia coli (Lewis, L. K., Jenkins, M. E., and Mount, D. W. (1992) J. Bacteriol. 174, 3377-3385). Sequence analysis suggests that the dinG gene encodes a putative helicase related to a group of eukaryotic helicases that includes mammalian XPD (Koonin, E. V. (1993) Nucleic Acids Res. 21, 1497), an enzyme involved in transcription-coupled nucleotide excision repair and basal transcription. We have characterized the dinG gene product from E. coli using genetic and biochemical approaches. Deletion of dinG has no severe phenotype, indicating that it is non-essential for cell viability. Both dinG deletion and over-expression of the DinG protein from a multicopy plasmid result in a slight reduction of UV resistance. DinG, purified as a fusion protein from E. coli cells, behaves as a monomer in solution, as judged from gel filtration experiments. DinG is an ATP-hydrolyzing enzyme; single-stranded (ss) DNA stimulates the ATPase activity 15-fold. Kinetic data yield a Hill coefficient of 1, consistent with one ATP-hydrolyzing site per DinG molecule. DinG possesses a DNA helicase activity; it translocates along ssDNA in a 5' --> 3' direction, as revealed in experiments with substrates containing non-natural 5'-5' and 3'-3' linkages. The ATP-dependent DNA helicase activity of DinG requires divalent cations (Mg2+, Ca2+, and Mn2+) but is not observed in the presence of Zn2+. The DinG helicase does not discriminate between ribonucleotide and deoxyribonucleotide triphosphates, and it unwinds duplex DNA with similar efficiency in the presence of ATP or dATP. We discuss the possible involvement of the DinG helicase in DNA replication and repair processes. PMID- 12748190 TI - p21/CDKN1A mediates negative regulation of transcription by p53. AB - The tumor suppressor p53 regulates transcription positively and negatively, depending on the target gene. Whereas p53 induces transcription through direct interaction with promoter DNA, the mechanism of p53-mediated transcriptional repression is less well understood. Early reports described the alleviation of p53-mediated repression by inhibitors of apoptosis, suggesting that negative regulation of transcription might occur only in conjunction with programmed cell death. More recently, it has been proposed that certain genes, such as survivin, are repressed by direct association of p53 with their promoters, followed by recruitment of a repressor complex. We show here that p53-mediated negative regulation of transcription could occur independently of apoptosis. In contrast, the amino-terminal transactivation domain of p53 was required for negative regulation of transcription. Similarly, the p53 homologue p73 diminished the expression of survivin and stathmin, depending on its transactivation domain. Mutation of the putative p53 binding site within the survivin promoter did not impair its repression. These observations raised the hypothesis that activation of an effector gene might be required for repression by p53. Strikingly, when the p53-inducible p21/CDKN1A gene was deleted, p53 no longer repressed any one among 11 genes that it down-regulates otherwise. Most of these genes were also repressed by ectopic p21 in the absence of p53. Overexpressed c-Myc reduced the transcription of p21/CDKN1A and impaired p53-mediated repression but did not abolish repression by ectopic p21. Taken together, these results strongly suggest that increased expression of p21/CDKN1A is necessary and sufficient for the negative regulation of gene expression by p53. PMID- 12748191 TI - Saccharomyces cerevisiae PIP2 mediating oleic acid induction and peroxisome proliferation is regulated by Adr1p and Pip2p-Oaf1p. AB - Saccharomyces cerevisiae genes involved in fatty acid degradation contain in their promoters oleate response elements (OREs) and type 1 upstream activation sequences (UAS1s) that bind Pip2p-Oaf1p and Adr1p, respectively. The promoter of the PIP2 gene was found to contain a potential UAS1 that consists of a tandem array of CYCCRR half-sites in an overlapping arrangement with a previously characterized ORE. Electrophoretic mobility shift analysis demonstrated that Adr1p bound to UAS1PIP2, and Northern analysis in combination with a lacZ reporter gene confirmed that Adr1p influenced the transcription of PIP2. Immunoprecipitation showed that, in adr1delta mutant cells grown on oleic acid, Pip2p was less abundant compared with the corresponding wild-type. In addition, the amount of Pip2p-Oaf1p that bound to a target ORE in vitro was reduced in mutant extracts compared with the wild-type. Transcription of the oleic acid inducible genes SPS19 and CTA1, which rely on both Pip2p-Oaf1p and Adr1p for their regulation, was reduced in adr1delta mutant cells. However, by ectopically restoring levels of Pip2p in adr1delta cells grown on oleic acid medium, transcription of both genes increased 2-fold compared with the control. This partial suppression of the adr1delta mutant phenotype was additionally manifested by moderate utilization of oleic acid. Hence, both the expression as well as the action of the two transcription factors, Adr1p and Pip2p-Oaf1p, are interconnected, which allows for an elaborate control of fatty acid-inducible genes. PMID- 12748193 TI - The role of the SAP motif in promoting Holliday junction binding and resolution by SpCCE1. AB - Holliday junctions are four-way branched DNA structures that are formed during recombination and by replication fork regression. Their processing depends on helicases that catalyze junction branch migration, and endonucleases that resolve the junction into nicked linear DNAs. Here we have investigated the role of a DNA binding motif called SAP in binding and resolving Holliday junctions by the fission yeast mitochondrial resolvase SpCCE1. Mutation or partial/complete deletion of the SAP motif dramatically impairs the ability of SpCCE1 to resolve Holliday junctions in a heterologous in vivo system. These mutant proteins retain the ability to recognize the junction structure and to distort it upon binding. However, once formed the mutant protein-junction complexes are relatively unstable and dissociate much faster than wild-type complexes. We show that binding stability is necessary for efficient junction resolution, and that this may be due in part to a requirement for maintaining the junction in an open conformation so that it can branch migrate to cleavable sites. PMID- 12748192 TI - Phosphorylation of threonine 10 on CKBBP1/SAG/ROC2/Rbx2 by protein kinase CKII promotes the degradation of IkappaBalpha and p27Kip1. AB - In eukaryotic cells, protein kinase CKII is required for progression through the cell division cycle. We recently reported that CKBBP1/SAG/ROC2/Rbx2 associates with the beta-subunit of CKII and is phosphorylated by purified CKII in the presence of ATP in vitro. In this report, we demonstrate that CKBBP1 is efficiently phosphorylated in vitro by purified CKII in the presence of GTP and by heparin-sensitive protein kinase in HeLa cell extract. Mutational analysis indicates that CKII phosphorylates threonine at residue 10 within CKBBP1. Furthermore, CKBBP1 is phosphorylated in vivo and threonine to alanine mutation at residue 10 abrogates the phosphorylation of CKBBP1 observed in vivo, indicating that CKII is a major kinase that is responsible for in vivo phosphorylation of CKBBP1. As compared with the wild-type CKBBP1 or CKBBP1T10E (in which threonine 10 is replaced by glutamate), overexpression of nonphosphorylatable CKBBP1 (CKBBP1T10A) results in accumulation of IkappaBalpha and p27Kip1. Experiments using proteasome inhibitor MG132 and CKII inhibitor 5,6 dichloro-1-beta-d-ribofuranosylbenzimidazole suggest that the accumulation of IkappaBalpha and p27Kip1 results primarily from the reduction of proteasomal degradation in cells expressing CKBBP1T10A, and that CKII-mediated CKBBP1 phosphorylation is required for efficient degradation of IkappaBalpha and p27Kip1. Overexpression of CKBBP1T10A in HeLa cells suppresses cell proliferation and causes accumulation of G1/G0 peak of the cell cycle. Taken together, our results indicate that CKII may control IkappaBalpha and p27Kip1 degradation and thereby G1/S phase transition through the phosphorylation of threonine 10 within CKBBP1. PMID- 12748194 TI - Regulation of the mhp cluster responsible for 3-(3-hydroxyphenyl)propionic acid degradation in Escherichia coli. AB - The mhp gene cluster from Escherichia coli constitutes a model system to study bacterial degradation of 3-(3-hydroxyphenyl)propionic acid (3HPP). In this work the regulation of the inducible mhp catabolic genes has been studied by genetic and biochemical approaches. The Pr and Pa promoters, which control the expression of the divergently transcribed mhpR regulatory gene and mhp catabolic genes, respectively, show a peculiar arrangement leading to transcripts that are complementary at their 5'-ends. By using Pr-lacZ and Pa-lacZ translational fusions and gel retardation assays, we have shown that the mhpR gene product behaves as a 3HPP-dependent activator of the Pa promoter, being the expression from Pr constitutive and MhpR-independent. DNase I footprinting experiments and mutational analysis mapped an MhpR-protected region, centered at position -58 with respect to the Pa transcription start site, which is indispensable for MhpR binding and in vivo activation of the Pa promoter. Superimposed in the specific MhpR-mediated regulation of the Pa promoter, we have observed a strict catabolite repression control carried out by the cAMP receptor protein (CRP) that allows expression of the mhp catabolic genes when the preferred carbon source (glucose) is not available and 3HPP is present in the medium. Gel retardation assays revealed that the specific activator, MhpR, is essential for the binding of the second activator, CRP, to the Pa promoter. Such peculiar synergistic transcription activation has not yet been observed in other aromatic catabolic pathways, and the MhpR activator becomes the first member of the IclR family of transcriptional regulators that is indispensable for recruiting CRP to the target promoter. PMID- 12748195 TI - Characterizing the structural features of RNA/RNA interactions of the F-plasmid FinOP fertility inhibition system. AB - F-like plasmid transfer is mediated by the FinOP fertility inhibition system. Expression of the F positive regulatory protein, TraJ, is controlled by the action of the antisense RNA, FinP, and the RNA-binding protein FinO. FinO binds to and protects FinP from degradation and promotes duplex formation between FinP and traJ mRNA, leading to repression of both traJ expression and conjugative F transfer. FinP antisense RNA secondary structure is composed of two stem-loops separated by a 4-base single-stranded spacer and flanked on each side by single stranded tails. Here we show that disruption of the expected Watson-Crick base pairing between the loops of FinP stem-loop I and its cognate RNA binding partner, traJ mRNA stem-loop Ic, led to a moderate reduction in the rate of duplex formation in vitro. In vivo, alterations of the anti-ribosome binding site region in the loop of FinP stem-loop I reduced the ability of the mutant FinP to mediate fertility inhibition and to inhibit TraJ expression when expressed in trans at an elevated copy number. Alterations of intermolecular complementarity between the stems of these RNAs reduced the rate of duplex formation. Our results suggest that successful interaction between stem-loop I of FinP and stem-loop Ic of traJ mRNA requires that base pairing must proceed from an initial loop-loop interaction through the top portion of the stems for stable duplex formation to occur. PMID- 12748196 TI - Functional characterization and crystal structure of the C215D mutant of protein tyrosine phosphatase-1B. AB - We have characterized the C215D active-site mutant of protein-tyrosine phosphatase-1B (PTP-1B) and solved the crystal structure of the catalytic domain of the apoenzyme to a resolution of 1.6 A. The mutant enzyme displayed maximal catalytic activity at pH approximately 4.5, which is significantly lower than the pH optimum of 6 for wild-type PTP-1B. Although both forms of the enzyme exhibited identical Km values for hydrolysis of p-nitrophenyl phosphate at pH 4.5 and 6, the kcat values of C215D were approximately 70- and approximately 7000-fold lower than those of wild-type PTP-1B, respectively. Arrhenius plots revealed that the mutant and wild-type enzymes displayed activation energies of 61 +/- 1 and 18 +/- 2 kJ/mol, respectively, at their pH optima. Unlike wild-type PTP-1B, C215D mediated p-nitrophenyl phosphate hydrolysis was inactivated by 1,2-epoxy-3-(p nitrophenoxy)propane, suggesting a direct involvement of Asp215 in catalysis. Increasing solvent microviscosity with sucrose (up to 40% (w/v)) caused a significant decrease in kcat/Km of the wild-type enzyme, but did not alter the catalytic efficiency of the mutant protein. Structurally, the apoenzyme was identical to wild-type PTP-1B, aside from the flexible WPD loop region, which was in both "open" and "closed" conformations. At physiological pH, the C215D mutant of PTP-1B should be an effective substrate-trapping mutant that can be used to identify cellular substrates of PTP-1B. In addition, because of its insensitivity to oxidation, this mutant may be used for screening fermentation broth and other natural products to identify inhibitors of PTP-1B. PMID- 12748197 TI - p38 Mitogen-activated protein kinase regulation of JB6 Cl41 cell transformation promoted by epidermal growth factor. AB - The relationship between cell transformation and p38 MAP kinase, a major mitogen activated protein (MAP) kinase pathway converting signals of various extracellular stimuli into expression of specific target genes through activation of transcription factors, still remains unclear. The aim of the present study was to investigate the role of the p38 MAP kinase pathway in epidermal growth factor (EGF)-induced cell transformation in JB6 cells. Our data show that a dominant negative mutant of p38 MAP (DN-p38) kinase inhibits EGF-promoted JB6 Cl41 cell transformation and that SB202190, an inhibitor of p38 MAP kinase, also inhibits JB6 Cl41 cell transformation in a dose-dependent manner. Moreover, our results show that DN-p38 MAP kinase inhibits the phosphorylation of EGF-stimulated activating transcription factor-2 (ATF-2) and signal transducer and activator of transcription 1 (STAT1). Additionally, DN-p38 MAP kinase inhibits EGF-induced phosphorylation of c-Myc (Thr58/Ser62). Gel shift assays indicate that DN-p38 MAP kinase inhibits EGF-induced activator protein-1 (AP-1) DNA binding in a dose dependent manner. These results show that p38 MAP kinase plays a key role in the regulation of EGF-induced cell transformation in JB6 cells through regulation of phosphorylation of p38 MAP kinase and activation of its target genes in phosphorylation, c-Myc cell transformation-related genes, and AP-1 binding ability. PMID- 12748198 TI - A single-stranded DNA-binding protein of bacteriophage T7 defective in DNA annealing. AB - The annealing of complementary strands of DNA is a vital step during the process of DNA replication, recombination, and repair. In bacteriophage T7-infected cells, the product of viral gene 2.5, a single-stranded DNA-binding protein, performs this function. We have identified a single amino acid residue in gene 2.5 protein, arginine 82, that is critical for its DNA annealing activity. Expression of gene 2.5 harboring this mutation does not complement the growth of a T7 bacteriophage lacking gene 2.5. Purified gene 2.5 protein-R82C binds single stranded DNA with a greater affinity than the wild-type protein but does not mediate annealing of complementary strands of DNA. A carboxyl-terminal-deleted protein, gene 2.5 protein-Delta26C, binds even more tightly to single-stranded DNA than does gene 2.5 protein-R82C, but it anneals homologous strands of DNA as well as does the wild-type protein. The altered protein forms dimers and interacts with T7 DNA polymerase comparable with the wild-type protein. Gene 2.5 protein-R82C condenses single-stranded M13 DNA in a manner similar to wild-type protein when viewed by electron microscopy. PMID- 12748199 TI - The Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure: the JNC 7 report. AB - "The Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure" provides a new guideline for hypertension prevention and management. The following are the key messages(1) In persons older than 50 years, systolic blood pressure (BP) of more than 140 mm Hg is a much more important cardiovascular disease (CVD) risk factor than diastolic BP; (2) The risk of CVD, beginning at 115/75 mm Hg, doubles with each increment of 20/10 mm Hg; individuals who are normotensive at 55 years of age have a 90% lifetime risk for developing hypertension; (3) Individuals with a systolic BP of 120 to 139 mm Hg or a diastolic BP of 80 to 89 mm Hg should be considered as prehypertensive and require health-promoting lifestyle modifications to prevent CVD; (4) Thiazide-type diuretics should be used in drug treatment for most patients with uncomplicated hypertension, either alone or combined with drugs from other classes. Certain high-risk conditions are compelling indications for the initial use of other antihypertensive drug classes (angiotensin-converting enzyme inhibitors, angiotensin-receptor blockers, beta-blockers, calcium channel blockers); (5) Most patients with hypertension will require 2 or more antihypertensive medications to achieve goal BP (<140/90 mm Hg, or <130/80 mm Hg for patients with diabetes or chronic kidney disease); (6) If BP is more than 20/10 mm Hg above goal BP, consideration should be given to initiating therapy with 2 agents, 1 of which usually should be a thiazide-type diuretic; and (7) The most effective therapy prescribed by the most careful clinician will control hypertension only if patients are motivated. Motivation improves when patients have positive experiences with and trust in the clinician. Empathy builds trust and is a potent motivator. Finally, in presenting these guidelines, the committee recognizes that the responsible physician's judgment remains paramount. PMID- 12748200 TI - JNC 7--it's more than high blood pressure. PMID- 12748201 TI - Heart failure: a global disease requiring a global response. PMID- 12748202 TI - An unusual case of chest murmur demonstrated with three dimensional volume rendering with 16 row multislice spiral computed tomography. PMID- 12748203 TI - Estimating prognosis in heart failure: time for a better approach. PMID- 12748205 TI - Drug treatment of hypertension: implications of ALLHAT. PMID- 12748207 TI - Non-invasive coronary imaging: computed tomography or magnetic resonance imaging? PMID- 12748208 TI - Multiple coronary rupture after blunt chest trauma. PMID- 12748209 TI - Congenital coronary artery dilatation. AB - Although dilated coronary arteries are uncommon in children, three cases are presented which, even more unusually, were not associated with Kawasaki's disease PMID- 12748211 TI - A thermal view of the arterial wall after coronary angioplasty. PMID- 12748210 TI - Microarray analysis: a novel research tool for cardiovascular scientists and physicians. AB - The massive increase in information on the human DNA sequence and the development of new technologies will have a profound impact on the diagnosis and treatment of cardiovascular diseases. The microarray is a micro-hybridisation based assay. The filter, called microchip or chip, is a special kind of membrane in which are spotted several thousands of oligonucleotides of cDNA fragments coding for known genes or expressed sequence tags. The resulting hybridisation signal on the chip is analysed by a fluorescent scanner and processed with a software package utilising the information on the oligonucleotide or cDNA map of the chip to generate a list of relative gene expression. Microarray technology can be used for many different purposes, most prominently to measure differential gene expression, variations in gene sequence (by analysing the genome of mutant phenotypes), or more recently, the entire binding site for transcription factors. Measurements of gene expression have the advantage of providing all available sequence information for any given experimental design and data interpretation in pursuit of biological understanding. This research tool will contribute to radically changing our understanding of cardiovascular diseases. PMID- 12748212 TI - Predicting mortality in patients with heart failure: a pragmatic approach. AB - OBJECTIVE: To develop a comprehensive and easily applicable prognostic model predicting mortality risk in patients with moderate to severe heart failure. DESIGN: Prospective follow up study. SETTING: Seven general hospitals in the Netherlands. PATIENTS: 152 outpatients with heart failure or patients admitted to hospital because of heart failure, who were included in a randomised trial to assess the impact of a pharmacist led intervention to improve drug compliance. Duration of follow up was at least 18 months. MAIN OUTCOME MEASURES: Multivariable logistic regression modelling was used to evaluate information from history, physical examination (for example, blood pressure), drug use, and quality of life questionnaires that independently contributed to the prediction of death. The area under receiver operating characteristic curves (AUC) was used to estimate the predictive ability of the prognostic models. RESULTS: During the 18 months of follow up, 51 patients (34%) died. Independent predictors of mortality were diabetes mellitus, a history of renal dysfunction (or higher creatinine), New York Heart Association (NYHA) functional class III or IV, lower weight or body mass index, lower blood pressure, ankle oedema, and higher scores on a disease specific quality of life questionnaire. The use of beta blockers was predictive of a better prognosis. These factors were used to derive various prediction formulas. A model based on medical history, weight, presence of oedema, and lower blood pressure had an AUC of 0.77. Addition of use of beta blockers to this model improved the AUC to 0.80. Addition of NYHA class increased the AUC to 0.84. Data on quality of life did not improve the AUC further (AUC 0.85). CONCLUSIONS: A prognostic model produced on the basis of easily obtainable information from medical history and physical examination can adequately stratify heart failure patients according to their short term risk of death. PMID- 12748213 TI - Pattern of ventilation during exercise in chronic heart failure. AB - OBJECTIVE: To determine the pattern of the abnormal ventilatory response in heart failure and how it relates to symptoms by looking at tidal volume (VT) and frequency (f) during exercise. METHODS: 45 patients with heart failure and 21 controls underwent maximal treadmill based exercise testing with metabolic gas exchange analysis. The relation of ventilation (VE) to VT was plotted to look for an inflection point where VT failed to increase further. The slope of the relation before this inflection point was documented. Time to the inflection point, VT, and f at the inflection point were recorded. The relation of symptom scores to f and E was also examined. RESULTS: Peak oxygen consumption (PVO2) (mean (SD)) was lower (19.7 (4.5) v 37.9 (8.6) ml/kg/min; p < 0001) and the ventilation to carbon dioxide production (VE/VCO2) slope was steeper (40.0 (6.5) v 26.0 (1.6); p < 0.0001) in patients with heart failure than in the control group. The patients reached the inflection point of the VE/VT slope sooner during exercise than the controls (271 (110) v 502 (196) seconds; p < 0.0001). Patients had a higher f and a smaller VT at that point and throughout exercise until the peak where f was the same for patients and controls. VT at the inflection point correlated with PVO2 (r = 0.67; p < 0.0001). Despite having an increased sensation of breathlessness for a given E, patients were less symptomatic of f than controls. CONCLUSIONS: Patients with heart failure breathe at a higher f throughout exercise, reaching an apparent maximal VT earlier. The VT at an inflection point on the VE/VT slope predicts PVO2. PMID- 12748214 TI - Prognosis for patients newly admitted to hospital with heart failure: survival trends in 12 220 index admissions in Leicestershire 1993-2001. AB - OBJECTIVE: To examine rates of, prognosis following, and the influences on first hospital admission with heart failure in Leicestershire during 1993-2001. DESIGN: Historical cohort study using record linked discharge and mortality data. SETTING: Leicestershire, England. PATIENTS: 12 220 individual patients newly hospitalised with heart failure between 1 April 1993 and 31 March 2001. MAIN OUTCOME MEASURES: 30 day and one year survival, temporal trends in survival, and the influence on prognosis of age, sex, comorbidity, social deprivation, and year of hospital admission. METHODS AND RESULTS: Between 1993/94 and 2000/01, rates of first hospitalisation increased by 62%, from 29 to 47/10 000 population, confined largely to those aged > 65 years. Rates did not increase after 1998. Median age at presentation increased from 74 years in 1993/94 to 77 years in 2000/01 for men but was unchanged (80 years) for women. Overall one and five year survival was 57% and 27%, respectively. There was a 43-45% increase in risk of death for each decade of age at admission and a 14-17% increase associated with male sex. There was a clear influence on outcome of comorbidity but no influence of social deprivation score. Both one month and one year survival were lower for patients whose first heart failure admission was concomitant with acute myocardial infarction. Between 1993/94 and 2000/01 postdischarge cardiovascular survival improved by 50% (p < 0.001). CONCLUSIONS: Rates of first hospital admission with heart failure reached a plateau in the late 1990s. Case fatality rates remain high and prognosis poor, in particular for those of increasing age, for men, and for patients with concomitant acute myocardial infarction. However, clear trends to improved survival were seen over this time. PMID- 12748215 TI - Different responses to dobutamine in the presence of carvedilol or metoprolol in patients with chronic heart failure. AB - OBJECTIVE: To determine whether patients with congestive heart failure on different beta adrenoreceptor blocking drugs have similar haemodynamic responses to dobutamine. DESIGN: Single centre, single blind, randomised, two period crossover study comparing carvedilol with metoprolol CR/XL. PATIENTS: Ten patients with stable chronic congestive heart failure (ejection fraction < 40%) on chronic treatment with metoprolol CR/XL. METHODS: Patients were treated with carvedilol or metoprolol CR/XL (target dose 50 mg twice daily and 200 mg once daily, respectively) for eight weeks. Stress echocardiography was undertaken at the end of each maintenance period, using dobutamine 5 and 15 microg/kg/min. RESULTS: No significant haemodynamic differences were seen at rest on the two treatments. There was a more pronounced increase in heart rate and cardiac output during dobutamine infusion when the patients were on metoprolol than when they were on carvedilol. Mean arterial pressure increased significantly when the patients were on carvedilol, and cardiac output increased during low dose dobutamine, without further change during high dose dobutamine. During the dobutamine infusion, there was no significant difference in ejection fraction between carvedilol and metoprolol treatment. CONCLUSIONS: Patients with congestive heart failure on a non-selective beta adrenoreceptor blocker or beta1 selective blocker responded differently to the inotropic drug dobutamine: the beta1 blockade caused by metoprolol could be counteracted by dobutamine, whereas with carvedilol a low dose of dobutamine increased cardiac output, and a higher dose of dobutamine caused a pressor effect. These findings may be clinically relevant when choosing an inotropic drug. PMID- 12748216 TI - Coronary calcifications in young patients with first, unheralded myocardial infarction: a risk factor matched analysis by electron beam tomography. AB - OBJECTIVE: To compare the presence and extent of coronary calcifications in young patients with first, unheralded acute myocardial infarction with matched controls without a history of coronary artery disease. METHODS: In 102 patients under 60 years of age (19-59 years, mean 41 years; 88% male), electron beam tomography was done 1-14 days after acute myocardial infarction, before any coronary intervention. Coronary calcifications were quantified using the Agatston score. Age related calcium centiles were determined based on the Mayo Clinic "epidemiology of coronary calcification" study, and results were compared with a group of 102 controls without coronary artery disease, matched for sex, age, and risk factors. RESULTS: Calcifications were present in 95.1% of patients with acute myocardial infarction and in 59.1% of controls (p = 0.008). The mean (SD) Agatston score was 529 (901) in the infarct patients versus 119 (213) in the controls (p < 0.001). An Agatston score above the 50th centile was present in 87.2% of infarct patients and 47.0% of controls (p = 0.006), and above the 90th centile in 60.7% of infarct patients and only 5.8% of controls (p = 0.001). CONCLUSIONS: In young patients with their first, unheralded acute myocardial infarction, the presence and extent of coronary calcium are significantly greater than in matched controls. PMID- 12748217 TI - Comparative follow up of patients with implanted cardioverter-defibrillators after induction of sustained monomorphic ventricular tachycardias or ventricular fibrillation by programmed stimulation. AB - OBJECTIVE: To investigate the prognostic value of induced monomorphic ventricular tachycardia (VT) and ventricular flutter or fibrillation (VF) during programmed electrical stimulation in patients with a high risk for sudden arrhythmogenic cardiac death. DESIGN: Prospective cohort study. PATIENTS: 102 patients at high risk for arrhythmogenic sudden cardiac death who received an automated implantable cardioverter-defibrillator (AICD) were evaluated. 56 patients received the AICD for primary prevention and 46 for secondary prevention. 58 patients had induction of a monomorphic VT (VT group) and 44 had induction of a polymorphic VT, ventricular flutter, or ventricular fibrillation (VF group) during programmed electrical stimulation. Average follow up was 20 months in both groups. MAIN OUTCOME MEASURES: Appropriate AICD protocol. RESULTS: In patients who received the AICD for primary prevention, 16 of 32 patients in the VT group, compared with only four of 24 patients in the VF group, received an appropriate AICD protocol (p = 0.02). In the entire study population, 479 appropriate AICD protocols were recorded in 28 (48%) patients in the VT group and 28 appropriate protocols in 11 (25%) patients in the VF group. Cumulative Kaplan-Meier event free survival curves were significantly different (p = 0.02). CONCLUSION: Induction of VF during programmed electrical stimulation is of no prognostic value even in high risk patients without previously documented ventricular fibrillation. PMID- 12748218 TI - Non-invasive intravenous coronary angiography using electron beam tomography and multislice computed tomography. AB - BACKGROUND: Electron beam computed tomography (EBCT) and multislice computed tomography (MSCT) are both suitable for non-invasive identification of coronary stenoses. OBJECTIVE: To compare intravenous coronary EBCT angiography (EBCTA) and MSCT angiography (MSCTA) with regard to image quality and diagnostic accuracy. METHODS: EBCTA was done using an Imatron C-150 XP scanner in 101 patients following a standard protocol (slice thickness 3 mm, overlap 1 mm, acquisition time 100 ms, prospective ECG trigger). For MSCTA in a different set of 91 patients (using a Siemens Somatom Plus4VZ scanner), the whole volume of the heart was covered in a spiral technique by four simultaneous detector rows. Using retrospective ECG gating, the raw data were reconstructed in (mean (SD)) 215 (12) axial slices acquired in diastole (slice thickness 1.25 mm, overlap 0.5 mm, acquisition time 250 ms/slice). RESULTS: With EBCTA, 76% of predetermined coronary segments in a nine segment model could be assessed with diagnostic image quality, and with MSCTA, 82%. A low contrast to noise ratio with EBCTA, and the presence of motion artefacts with MSCTA were the main reasons for inadequate image quality. Using conventional angiography as the gold standard, 77% of stenoses of > 50% could be identified correctly with EBCTA and 82% with MSCTA. Significant stenoses were correctly ruled out in 93% of segments with EBCTA, and in 96% of segments with MSCTA. The average contrast to noise ratio was higher with MSCTA than with EBCTA (9.4 v 6.5; p < 0.001). CONCLUSIONS: EBCTA and MSCTA show similarly high levels of accuracy for determining and ruling out significant coronary artery stenoses. MSCTA is capable of providing good image quality in more coronary segments than EBCTA because of its better contrast to noise ratio and higher spatial resolution. Motion artefacts seen at heart rates of > 75 beats/min and a higher radiation exposure are the main limitations of MSCTA. PMID- 12748219 TI - Fibromuscular dysplasia in renovascular hypertension demonstrated by multislice CT: comparison with conventional angiogram and intravascular ultrasound. PMID- 12748220 TI - Clinical and angiographic acute and follow up results of intracoronary beta brachytherapy in saphenous vein bypass grafts: a subgroup analysis of the multicentre European registry of intraluminal coronary beta brachytherapy (RENO). AB - OBJECTIVE: To assess clinically and angiographically the feasibility, safety, and effectiveness of vascular brachytherapy (VBT) in saphenous vein bypass grafts (SVG). PATIENTS AND METHODS: 67 of 1098 (6.1%) consecutive patients of the European registry of intraluminal coronary beta brachytherapy underwent treatment for 68 SVG lesions by VBT using a Sr/Y(90) source train (BetaCath). Clinical follow up data were obtained for all of them after a mean (SD) of 6.3 (2.4) months and angiographic follow up was performed in 61 patients (91.0%) after 6.9 (2.0) months. RESULTS: 58 (86.6%) patients were men, their mean (SD) age was 66 (10) years, 28 (41.8%) had unstable angina, and 21 (31.3%) had diabetes. Fifty three (77.9%) lesions were in-stent restenosis, 13 (19.1%) de novo lesions, and 2 (3.0%) non-stented restenotic lesions. Mean (SD) reference diameter before the intervention was 4.19 (0.52) mm, mean (SD) lesion length was 23.56 (20.38) mm, and mean (SD) minimum lumen diameter measured 0.73 (0.62) mm. Mean (SD) acute gain was 3.02 (0.88) mm. The prescribed radiation dose was 20.1 (3.2) Gy. Pullback manoeuvres were performed in 17 (25.0%) of cases. Most patients received combined aspirin and thienopyridin treatment for 6 or 12 months after the procedure. Technical success was obtained in 62 (91.2%) treated lesions and in hospital major adverse cardiac events occurred in 4.5%. At follow up, mean (SD) reference diameter was 4.20 (0.53) mm, minimum lumen diameter 2.94 (1.50) mm, and late loss 0.86 (1.25) mm. The overall major adverse cardiac events rate was 26.7%. CONCLUSION: VBT of SVG is feasible and safe. At follow up the reintervention rate and cardiac morbidity and mortality seem to be favourable, considering that interventions in SVG usually are associated with the highest risks. PMID- 12748221 TI - Left ventricular microaneurysms as a cause of apparently idiopathic ventricular fibrillation. PMID- 12748222 TI - Stenting of the ductus arteriosus and banding of the pulmonary arteries: basis for various surgical strategies in newborns with multiple left heart obstructive lesions. AB - OBJECTIVE: To present an institutional experience with stent placement in the arterial duct combined with bilateral banding of the pulmonary artery branches as a basis for various surgical strategies in newborns with hypoplastic left heart obstructive lesions. DESIGN: Observational study. SETTING: Paediatric heart centre in a university hospital. PATIENTS: 20 newborns with various forms of left heart obstructive lesions and duct dependent systemic blood flow. INTERVENTIONS: Patients underwent percutaneous ductal stenting and surgical bilateral pulmonary artery banding. Atrial septotomy by balloon dilatation was performed as required, in one premature baby by the transhepatic approach. MAIN OUTCOME MEASURES: Survival; numbers of and reasons for palliative and corrective cardiac surgery. RESULTS: One patient died immediately after percutaneous ductal stenting. One patient died in connection with the surgical approach of bilateral pulmonary banding. Stent and ductal patency were achieved for up to 331 days. Two patients underwent heart transplantation and two patients died on the waiting list. Ten patients had a palliative one stage procedure with reconstruction of the aortic arch and bidirectional cavopulmonary connection at the age of 3.5-6 months. There was one death. One patient is still awaiting this approach. Two patients received biventricular repair. In one, biventricular repair will soon be provided. CONCLUSIONS: Stenting the arterial duct combined with bilateral pulmonary artery banding in newborns with hypoplastic left heart or multiple left heart obstructive lesions allows a broad variation of surgical strategies depending on morphological findings, postnatal clinical conditions, and potential ventricular growth. PMID- 12748223 TI - Thirty years after percutaneous closure of patent ductus arteriosus. PMID- 12748224 TI - Biocorrosion of magnesium alloys: a new principle in cardiovascular implant technology? AB - OBJECTIVES: To develop and test a new concept of the degradation kinetics of newly developed coronary stents consisting of magnesium alloys. METHODS: Design of a coronary stent prototype consisting of the non-commercial magnesium based alloy AE21 (containing 2% aluminium and 1% rare earths) with an expected 50% loss of mass within six months. Eleven domestic pigs underwent coronary implantation of 20 stents (overstretch injury). RESULTS: No stent caused major problems during implantation or showed signs of initial breakage in the histological evaluation. There were no thromboembolic events. Quantitative angiography at follow up showed a significant (p < 0.01) 40% loss of perfused lumen diameter between days 10 and 35, corresponding to neointima formation seen on histological analysis, and a 25% re-enlargement (p < 0.05) between days 35 and 56 caused by vascular remodelling (based on intravascular ultrasound) resulting from the loss of mechanical integrity of the stent. Inflammation (p < 0.001) and neointimal plaque area (p < 0.05) depended significantly on injury score. Planimetric degradation correlated with time (r = 0.67, p < 0.01). CONCLUSION: Vascular implants consisting of magnesium alloy degradable by biocorrosion seem to be a realistic alternative to permanent implants. PMID- 12748225 TI - Left main coronary artery fistula exiting into the right atrium. PMID- 12748226 TI - Intramyocardial coronary flow characteristics in patients with hypertrophic cardiomyopathy: non-invasive assessment by transthoracic Doppler echocardiography. PMID- 12748227 TI - Measurements of cardiac troponin T in patients with hypertrophic cardiomyopathy. PMID- 12748228 TI - Brain natriuretic peptide facilitates severity classification of stable chronic heart failure with left ventricular dysfunction. PMID- 12748230 TI - Coronary anomaly imaging by multislice computed tomography in corrected tetralogy of Fallot. PMID- 12748229 TI - Adipocyte derived plasma protein, adiponectin, is associated with smoking status in patients with coronary artery disease. PMID- 12748231 TI - Same day discharge following elective percutaneous coronary intervention in patients with stable angina. PMID- 12748232 TI - Valved conduit from the left ventricular apex to ascending aorta for recurrent congenital subaortic stenosis: 25 years later. PMID- 12748233 TI - The variation of plasma concentrations of a novel, adipocyte derived protein, adiponectin, in patients with acute myocardial infarction. PMID- 12748234 TI - Early detection of left ventricular dysfunction in patients with beta thalassaemia major by dobutamine stress echocardiography. PMID- 12748235 TI - Role of exercise treadmill testing in the management of erectile dysfunction: a joint cardiovascular/erectile dysfunction clinic. PMID- 12748236 TI - Resuscitation. PMID- 12748237 TI - Ethnic differences in cardiovascular disease. PMID- 12748238 TI - A rational basis for selection among drugs of the same class. PMID- 12748240 TI - Transoesophageal echocardiography of a large tricuspid valve vegetation: a perfect image of reality. PMID- 12748241 TI - Novel use of an "insertable" loop recorder. AB - A patient with palpitations and suspected arrhythmia underwent Holter and external loop recorder monitoring. No arrhythmias were detected by these traditional monitoring methods. An insertable loop recorder (ILR) was placed on the patient's chest and used as an extended loop recorder. An arrhythmia was ultimately recorded by the externally placed ILR leading to appropriate treatment. PMID- 12748242 TI - A 28 year old woman with ventricular tachycardia and an abnormal chest radiograph. AB - A 28 year old white woman with no medical history presented to the emergency room with symptomatic non-sustained ventricular tachycardia. She was asymptomatic up to a few days before presentation. Her physical examination was essentially normal and hence did not help with the differential diagnosis of the problem. Bronchoscopic transbronchial biopsy led to the final diagnosis of cardiac sarcoidosis. PMID- 12748243 TI - Gefitinib in recurrent non-small-cell lung cancer: an IDEAL trial? PMID- 12748245 TI - Expression of mucins and cytokeratins in primary carcinomas of the digestive system. AB - To determine the most optimal treatment of cancer patients, it is fundamental to classify human carcinomas according to their primary anatomical site of origin. As for some patients, it is difficult to identify cancers occurring at obscure location and overlapping adjacent sites. The aim of this study is to partition the primary site of 486 patients in cancers of the digestive system by the expression pattern of the mucins and cytokeratins typifying each site. The expressions of MUC1, MUC2, MUC5AC, MUC6, CK7, CK8, CK13, CK14, CK18, CK19 and CK20 were evaluated immunohistochemically in 426 adenocarcinomas and 60 hepatocellular carcinomas using the tissue-array method. The finding of MUC series showed their characteristics in case of MUC2 in the appendix cancer and MUC1 and 5AC in pancreas cancer. As for CKs 7, 13, and 19, and 20 had a feature in cancers of common bile duct, liver, and appendix, respectively. We classified cancers in 11 sites by characteristic expression of antibodies. The sensitivity, specificity, positive predictive value, and diagnostic efficacy of significant antibodies were calculated with deducing the dichotomous tree made by SPSS 10.0. Six of 11 antibodies, CK 7, CK13, CK19, CK20, MUC1, and MUC5AC distinguished 6 groups from 11 sites. We also executed the clustering of cancers to investigate total relationship among cancers. They fell into three categories, which corresponded to embryologic origin. Unlike other sites, the small intestine and colorectum cancers expressed significantly different patterns to their sublocations. Mucins and CKs showed expression patterns to classify the primary sites of digestive cancers and may be helpful in predicting the primary sites of digestive cancers. PMID- 12748244 TI - Multi-institutional randomized phase II trial of gefitinib for previously treated patients with advanced non-small-cell lung cancer (The IDEAL 1 Trial) [corrected]. AB - PURPOSE: To evaluate the efficacy and tolerability of two doses of gefitinib (Iressa [ZD1839]; AstraZeneca, Wilmington, DE), a novel epidermal growth factor receptor tyrosine kinase inhibitor, in patients with pretreated advanced non small-cell lung cancer (NSCLC). PATIENTS AND METHODS: This was a randomized, double-blind, parallel-group, multicenter phase II trial. Two hundred ten patients with advanced NSCLC who were previously treated with one or two chemotherapy regimens (at least one containing platinum) were randomly assigned to receive either 250-mg or 500-mg oral doses of gefitinib once daily. RESULTS: Efficacy was similar for the 250- and 500-mg/d groups. Objective tumor response rates were 18.4% (95% confidence interval [CI], 11.5 to 27.3) and 19.0% (95% CI, 12.1 to 27.9); among evaluable patients, symptom improvement rates were 40.3% (95% CI, 28.5 to 53.0) and 37.0% (95% CI, 26.0 to 49.1); median progression-free survival times were 2.7 and 2.8 months; and median overall survival times were 7.6 and 8.0 months, respectively. Symptom improvements were recorded for 69.2% (250 mg/d) and 85.7% (500 mg/d) of patients with a tumor response. Adverse events (AEs) at both dose levels were generally mild (grade 1 or 2) and consisted mainly of skin reactions and diarrhea. Drug-related toxicities were more frequent in the higher-dose group. Withdrawal due to drug-related AEs was 1.9% and 9.4% for patients receiving gefitinib 250 and 500 mg/d, respectively. CONCLUSION: Gefitinib showed clinically meaningful antitumor activity and provided symptom relief as second- and third-line treatment in these patients. At 250 mg/d, gefitinib had a favorable AE profile. Gefitinib 250 mg/d is an important, novel treatment option for patients with pretreated advanced NSCLC [corrected] PMID- 12748246 TI - Morphology of the internal elastic lamina in arteries from pulmonary hypertensive patients: a confocal laser microscopy study. AB - The development and progression of pulmonary hypertension lesions involve continuous remodeling of the arterial wall, including the extracellular matrix components. The integrity of the internal elastic lamina may represent a barrier to cell migration and formation of intimal proliferative lesions. Some patients with congenital cardiac shunts develop precocious intimal occlusive lesions,whereas others evolve with isolated medial hypertrophy. We studied the 2 D and 3-D morphology of the internal elastic lamina of peripheral pulmonary arteries to search for any difference regarding the type of histological lesion. Fifteen lung biopsies collected for diagnostic purposes from patients with congenital shunts and 6 control lungs (mean ages, 15.8 and 14.7 mo) were studied using the confocal laser scanning microscope, under predetermined conditions of laser intensity, brightness and contrast. We measured the thickness of the internal elastic lamina and determined the number of gaps and projections of elastic tissue towards the medial and intimal layers. The mean internal elastic lamina thickness was significantly higher in arteries from cases with isolated medial hypertrophy when compared with controls and to those with proliferative lesions (P <.05). The number of gaps of the internal elastic lamina was higher in arteries >100 micro m in diameter from the group with intimal lesions when compared to the cases presenting with isolated hypertrophy, but did not differ from the controls. There was a positive linear correlation between the external arterial diameter and the thickness of the internal elastic lamina (r =.74, P <.001) in cases presenting isolated medial hypertrophy. The increased thickness and smaller number of gaps of the internal elastic lamina may act as a barrier that prevents smooth muscle cell migration in patients with pulmonary hypertension without intimal proliferative lesions. On the other hand, a greater number of gaps does not represent, by itself, unrestrained migration, because controls also showed fenestrated laminae. PMID- 12748247 TI - Histopathological and clinical evaluation of serrated adenomas of the colon and rectum. AB - We evaluated the diagnostic utility of the histological characteristics ascribed in the literature to serrated adenomas and developed a practical working model to allow their reliable identification. We also documented the frequency and location of serrated adenomas identified in an unselected series of individuals undergoing colonoscopic evaluation, as well as the clinical characteristics of those individuals. One hundred forty consecutive individuals (prospective polyp data set; 97 male, 43 female; age mean: 63.3 y; age range: 29-98 y) with 255 polyps were identified from 919 individuals undergoing colonoscopy. Further polyps previously removed from these individuals were added for the purpose of histological assessment (extended polyp data set, n = 380). All polyps were assessed by two independent examiners for eight selected architectural and cytological features of serrated adenomas. In the prospective polyp data set, 56 patients had 72 hyperplastic polyps, 7 had 9 serrated adenomas, 3 had 4 admixed polyps, and 98 had 170 conventional adenomas. There was no difference in the age, sex, or cancer association of the seven patients with serrated adenomas when compared with the case of other individuals with polyps. The prevalence of serrated adenomas was 9/919 (1%) in our population, with an average size of 5.8 mm. When assessing serrated adenomas histologically, the combination of nuclear dysplasia and serration of >/=20% of crypts provided the most accurate model for detection of these lesions (sensitivity 100%, specificity 97%). Other criteria provided supportive evidence but did not increase the diagnostic yield. The optimum model for the histological identification of the serrated adenoma includes the presence of a serrated architecture in >/=20% of crypts in association with surface epithelial dysplasia. PMID- 12748248 TI - Immunoblastic lymphoma in persons with AIDS-associated Kaposi's sarcoma: a role for Kaposi's sarcoma-associated herpesvirus. AB - Kaposi's sarcoma-associated herpesvirus, the viral agent of Kaposi's sarcoma, is associated with two lymphoproliferative disorders: primary effusion lymphoma and multicentric Castleman's disease. To identify other lymphoproliferative conditions linked with Kaposi's sarcoma-associated herpesvirus, we studied non Hodgkin's lymphomas arising in individuals with AIDS-associated Kaposi's sarcoma. Formalin-fixed tissues from 24 such lymphomas were examined. As expected, two primary effusion lymphomas were Kaposi's sarcoma-associated herpesvirus-positive, with immunohistochemistry demonstrating the Kaposi's sarcoma-associated herpesvirus latency-associated nuclear antigen in the nuclei of all neoplastic cells. Additionally, three of seven evaluable cases of the immunoblastic variant of diffuse large B-cell lymphoma (immunoblastic lymphoma) showed similar latency associated nuclear antigen staining. These Kaposi's sarcoma-associated herpesvirus-positive immunoblastic lymphomas resembled primary effusion lymphoma histologically but were not known to involve body cavities (sites included lymph nodes, soft tissues of the neck, and spleen). Notably, 5-20% of the neoplastic cells in the Kaposi's sarcoma-associated herpesvirus-positive immunoblastic lymphomas also showed cytoplasmic staining for viral interleukin-6, a biologically active cytokine homologue found in primary effusion lymphoma. We conclude that Kaposi's sarcoma-associated herpesvirus is present in some immunoblastic lymphomas in persons with AIDS-associated Kaposi's sarcoma. PMID- 12748249 TI - Hypermethylation of the p16 gene and lack of p16 expression in hepatoblastoma. AB - Hepatoblastoma is the most frequent pediatric liver tumor that develops mostly in young children. Abnormal regulation of cell cycle regulatory genes including p16 has been described, displaying no p16 mRNA and p16 protein in hepatoblastomas. The inactivation of p16, leading to the disruption of cell cycle control is involved in many types of human malignancies. However, the mechanism of the p16 inactivation in hepatoblastomas has not yet been elucidated. In this present study, we examined the methylation status of the p16 gene promoter by using methylation-specific PCR in 24 cases of hepatoblastomas and in 20 cases of corresponding non-neoplastic liver tissue. Aberrant methylation of 5' CpG islands of p16 was present in 12 of 24 (50.0%) cases of hepatoblastoma. Clinicopathologic parameters were not associated with the methylation status of p16. To correlate the methylation status of p16 with the expression of p16, immunohistochemical staining was done in tumors and non-neoplastic liver tissue. All non-neoplastic liver tissues displayed moderate, but heterogeneous immunoreactivity for p16. Eight of 12 (66.6%) methylation-positive hepatoblastomas showed a complete lack of immunoreactivity for p16. The other 4 methylation-positive hepatoblastomas had heterogeneous immunoreactivity. Nine of 12 (75.0%) unmethylated cases of hepatoblastoma displayed diffuse immunoreactivity, whereas 3 cases of unmethylated hepatoblastoma were not immunostained for p16. Our data indicate that the hypermethylation of p16 is a major mechanism of the transcriptional repression of p16 in hepatoblastomas, and we suggest that the inactivation of p16, leading to the lack of p16, may play an important role in the tumorigenesis of hepatoblastomas. PMID- 12748250 TI - Estrogen receptors (alpha and beta) and 17beta-hydroxysteroid dehydrogenase type 1 and 2 in thyroid disorders: possible in situ estrogen synthesis and actions. AB - Both epidemiological and experimental findings suggest the possible roles of sex steroids in the pathogenesis and/or development of various human thyroid disorders. In this study, we evaluated the expression of estrogen receptors (ER) alpha and beta in normal thyroid glands (N = 25; female: n = 13, male: n = 10, unknown: n = 2) ranging in age from fetus to adult. Furthermore, using immunohistochemistry, we investigated the expression of ERalpha and beta in 206 cases of thyroid disorders, including 24 adenomatous goiters, 23 follicular adenomas, and 159 thyroid carcinomas. In addition, we also studied the mRNA expression of ERalpha and beta and 17beta-hydroxysteroid dehydrogenase Type 1 and 2, enzymes involved in the interconversion between estrone and estradiol, using reverse transcription polymerase chain reaction (RT-PCR), in 48 of these 206 cases (10 adenomatous goiters, 10 follicular adenomas, and 28 papillary thyroid carcinomas) in which fresh frozen tissues were available for examination to further elucidate the possible involvement of intracrine estrogen metabolism and/or actions in thyroid disorders. ERalpha labeling index, or percentage of cells immunopositive for ERalpha, was significantly higher in adenomatous goiter (14.2 +/- 6.4), follicular adenoma (13.4 +/- 5.1), and thyroid carcinoma (16.4 +/ 2.1) than in normal thyroid gland (0; P <.05). Few follicular cells were positive for ERalpha in normal thyroid glands. In papillary carcinoma, ERalpha labeling index was significantly higher in premenopausal women (28.1 +/- 4.5) than in postmenopausal women (14.2 +/- 2.9) and in men of various ages (7.6 +/- 2.7; P <.05). In other histological types of thyroid carcinoma, no significant correlations were detected. ERbeta immunoreactivity was detected in both follicular and C-cells of normal thyroid glands, including those in developing fetal thyroid glands. In addition, ERbeta immunoreactivity was detected in the nuclei of various thyroid lesions. But no significant correlations were detected between ERbeta labeling index and clinicopathological findings including age, menopausal status, gender, and/or histological type of thyroid lesions. 17beta hydroxysteroid dehydrogenase Type 1 expression was detected in 31/48 (64.0%) of the cases examined, whereas Type 2 was detected only in 3/46 (6.3%) of all the cases examined. These results demonstrated that estrogens may influence the development, physiology, and pathology of human thyroid glands, and these effects, especially through ERalpha, may become more pronounced in neoplasms, particularly in papillary carcinoma arising in premenopausal women. PMID- 12748251 TI - Involvement of the chromosomal translocation t(11;18) in some mucosa-associated lymphoid tissue lymphomas and diffuse large B-cell lymphomas of the ocular adnexa: evidence from multiplex reverse transcriptase-polymerase chain reaction and fluorescence in situ hybridization on using formalin-fixed, paraffin-embedded specimens. AB - The chromosomal translocation t(11;18) is a unique chromosomal aberration associated with mucosa-associated lymphoid tissue lymphoma. API2 and MALT1 genes have been identified around this translocation. We attempted to find chromosomal abnormalities focusing mainly on the t(11;18) translocation in formalin-fixed, paraffin-embedded tissues of ocular adnexal lymphoproliferative disorders using multiplex reverse transcriptase-polymerase chain reaction and/or two-color interphase fluorescence in situ hybridization. By these methods, the t(11;18) translocation was detected in 1 of 8 patients with reactive lymphoid hyperplasia (13%), 3 of 23 with mucosa-associated lymphoid tissue lymphoma (13%), and 2 of 14 with diffuse large B-cell lymphoma with/without mucosa-associated lymphoid tissue lymphoma (14%). Moreover, we performed fluorescence in situ hybridization analysis to detect any numerical aberration of chromosomes 3, 7, 12, and 18 on some specimens nonselectively. No numerical chromosomal abnormalities were detected in 3 cases of reactive lymphoid hyperplasia, whereas three of four cases of mucosa-associated lymphoid tissue lymphoma and all four cases of diffuse large B-cell lymphoma with/without mucosa-associated lymphoid tissue lymphoma components exhibited one or more abnormalities. These findings indicate a possibility that at least in the ocular adnexa, some diffuse large B-cell lymphomas are derived from mucosa-associated lymphoid tissue lymphomas. PMID- 12748252 TI - Microtubule-associated protein-2 and class III beta-tubulin are expressed in extraskeletal myxoid chondrosarcoma. AB - Extraskeletal myxoid chondrosarcoma is a rare soft tissue sarcoma of uncertain histogenetic origin. Because recent reports have indicated neural-neuroendocrine differentiation in some extraskeletal myxoid chondrosarcomas, we investigated 25 tumors for expressions of microtubule-associated protein-2 and Class III beta tubulin, which are major components of microtubules and specifically localized in neurons and their derivatives. Immunohistochemical expression of microtubule associated protein-2 and Class III beta-tubulin was studied in extraskeletal myxoid chondrosarcomas using formalin-fixed, paraffin-embedded tissues. Cytoplasmic expressions of microtubule-associated protein-2 and Class III beta tubulin were detected in 21 (84%) and 13 (52%) of the 25 extraskeletal myxoid chondrosarcomas, respectively, although the number of positively stained tumor cells varied. Expression of the Class III beta-tubulin gene was also assessed in two immunohistochemically positive cases by in situ hybridization using an oligonucleotide probe specific for its transcript, and both cases showed expression of Class III beta-tubulin transcript. Another case was examined with immunoelectron microscopy, and immunogold particles for Class III beta-tubulin were localized to microtubular aggregates. Our data indicate that microtubules in extraskeletal myxoid chondrosarcoma are similar to those found in neurons, further supporting the concept that neural-neuroendocrine differentiation occurs in a significant number of extraskeletal myxoid chondrosarcoma. PMID- 12748254 TI - Increased expression of the bcl6 and CD10 proteins is associated with increased apoptosis and proliferation in diffuse large B-cell lymphomas. AB - There is increasing evidence that bcl6 and CD10 expression may be related to apoptosis and cell cycle progression. Therefore, 79 cases of de novo diffuse large B-cell lymphomas were studied for the expression of bcl6 and CD10 proteins in relation to 1) the apoptotic index; 2) the proliferation-associated proteins Ki67, cyclin A, and cyclin B1; and 3) the expression of the bcl2, p53, Rb, p16, and p27 proteins. Expression of bcl6, CD10, and bcl2 proteins was found in 54/79 (68%), 28/79 (35%), and 47/74 (63%) cases, respectively. The bcl6/CD10 patterns were as follows: bcl6+/CD10+ (26 cases, 32%), bcl6+/CD10- (28 cases, 33%), bcl6 /CD10- (23 cases, 31%), and bcl6-/CD10+ (2 cases, 4%). Significant positive correlations were found between bcl6/Ki67 (r =.328, P =.003), bcl6/cyclin A (r =.265, P =.018), bcl6/apoptotic index (r =.327, P =.010), CD10/Ki67 (r =.296, P =.008), and CD10/apoptotic index (r =.397, P =.001). In addition, high expression of bcl6 showed significant correlation with negative (null/low) bcl2 expression (chi(2) test, P =.002). The above findings indicate that increased expression of the bcl6 and CD10 proteins is associated with increased apoptosis and proliferation in diffuse large B-cell lymphomas. The association between increased bcl6 expression and enhanced apoptosis might be due, at least in part, to the null/low bcl2 expression because previous in vitro data showed that bcl6 overexpression induces apoptosis accompanied by bcl2 and bcl-xl downregulation. Moreover, significant correlation was found between increased apoptotic index and the bcl6+/CD10+ pattern (t test: P =.014, Mann-Whitney test: P =.046). This finding and the positive correlation of the apoptotic index with bcl6 and CD10 expression may be related to previous results showing that the expression of these proteins has favorable effects on the clinical outcome of diffuse large B cell lymphomas. PMID- 12748253 TI - Granzyme B in atherosclerosis and transplant vascular disease: association with cell death and atherosclerotic disease severity. AB - Apoptosis of intimal cells is an important contributor to the pathogenesis of atherosclerosis and transplant vascular disease (TVD). Since the activated immune response may be a key regulator of apoptosis in these lesions, we used immunohistochemistry to characterize the presence and localization of granzyme B, a major mediator of the cytotoxic immune response, in advanced atherosclerosis and TVD. Formalin-fixed, paraffin-embedded transverse sections from human left anterior descending coronary arteries were cut serially and stained with antibodies specific for granzyme B, smooth muscle alpha-actin, CD68, and CD3. The amount of granzyme B staining was semi-quantitated on a 0-5+/5+ scale. Also, TUNEL staining and in situ hybridization was performed to visualize cells undergoing cellular damage suggestive of apoptosis, and to localize granzyme B mRNA, respectively. Granzyme B localization was similar in both diseases. This protease was absent in arteries with mild atherosclerosis, but was abundant in the intima and media of vessels with advanced atherosclerosis and TVD. Within the intima, granzyme B localized to TUNEL-positive foam cells surrounding lipid-rich atheromas. Staining of serial sections with granzyme B and either smooth muscle alpha-actin, anti-CD68, or anti-CD3 showed that granzyme B localized to smooth muscle cells, macrophages, and T-cells. Further, in situ hybridization for granzyme B mRNA in TVD cases localized its expression to infiltrating leukocytes and not foam cells. In conclusion, the presence of granzyme B in advanced atherosclerotic lesions and TVD is associated with increasing disease severity and cell death. These observations suggest that granzyme B-mediated apoptosis may contribute to the pathogenesis of these diseases. PMID- 12748255 TI - PNL2, a new monoclonal antibody directed against a fixative-resistant melanocyte antigen. AB - We report the production of a new monoclonal antibody, PNL2, directed against a fixative resistant melanocyte antigen. The analysis of PNL2 immunostaining on a broad range of normal or malignant human tissues and on various melanocytic lesions revealed its high specificity. PNL2 gave a strong cytoplasmic staining of skin and oral mucosae melanocytes, and staining of granulocytes when used at high concentration. PNL2 stained all intra-epidermal nevi irrespective of their histologic type, but common intradermal nevi and the dermal component of compound nevi were largely non-reactive as only scattered nevus cells in the papillary dermis were labeled. PNL2 labeled more than 70% of the neoplastic cells in all primary melanomas irrespective of their histologic type. However, PNL2 did not label desmoplastic melanomas. All metastatic melanomas were also stained but the percentage of labeled cells was occasionally lower than the primary tumor. PNL2, as anti-Melan A and HMB-45 antibodies, stained most of the clear cell sarcoma cells, and a few cells in angiomyolipomas and lymphangioleiomyomatosis. None of the other non-melanocytic lesions tested were labeled. Proteomic approaches showed that the immunoaffinity purified PNL2-binding complexes isolated from melanoma cell lines comprise at least TAP1, Clathrin 17 and prealbumin proteins, but not the gp100 recognized by HMB-45. In conclusion, this new monoclonal antibody, PNL2, is directed against a new fixative resistant melanocyte associated antigen. This antigen is chemically resistant and thus allows immunostaining after melanin bleaching or decalcification. We also demonstrate that it is different from Melan A and from gp100, even if PNL2 and HMB-45 staining patterns are sometimes similar. PMID- 12748256 TI - Prognostic values of galectin-3 and the macrophage migration inhibitory factor (MIF) in human colorectal cancers. AB - This study aims to investigate whether the immunohistochemical levels of expression of galectin-3 and the macrophage migration inhibitory factor (MIF) are associated with prognostic values in human colorectal tumors. This was performed on 99 specimens including 69 colorectal tumors (17 Dukes A, 19 Dukes B, 15 Dukes C and 18 metastatic tumors that we labeled as D), 10 hepatic metastases from colorectal cancers and 20 normal specimens (biopsies). The immunohistochemical levels of expression of MIF and galectin-3 were quantified on routine histological slides by means of computer-assisted microscopy. Separate analyses were performed on epithelial and connective tissue. The levels of expression of both MIF and galectin-3 were very significantly higher in epithelial tumor tissue when compared with normal epithelial specimens. A positive and significant correlation between MIF and galectin-3 expression was evidenced in connective tumor tissue, and in particular in the cases associated with short survival periods (less than 5 years). In the case of the Dukes A or B tumors, we established two new prognostic groups (labeled I and II) on the basis of the levels of galectin-3 expression measured in the tumor epithelium. In the case of the Dukes C or D tumors, we established two other prognostic groups (labeled III and IV) on the basis of the levels of MIF expression measured in the connective tissue. Kaplan-Meyer analyses confirmed the additional prognostic values (as compared with conventional clinical staging) given by this new classification (groups I to IV). They show that the Dukes A or B tumors characterized by low levels of galectin-3 expression in the tumor epithelium are associated with significantly better prognoses than those characterized by high levels. In addition, the Dukes C or D tumors characterized by high levels of MIF expression in the connective tumor tissue are associated with significantly better prognoses than those characterized by low levels. In conclusions, MIF and galectin-3 expression levels in colorectal tumors are related to their levels of biological aggressiveness. These markers could be used to identify patients at risk, for whom more aggressive adjuvant therapy seems to be indicated. PMID- 12748257 TI - S100A6 protein expression is different in Spitz nevi and melanomas. AB - The Spitz nevus is a benign melanocytic lesion that can be identified reliably in many cases by conventional histopathological criteria. However, there are subsets of Spitz nevi and of malignant melanoma that closely resemble each other and represent diagnostic challenges. S100 proteins are of interest because of their involvement in neoplastic processes and their genes are clustered in chromosome 1q21. Chromosome 1 contains mutations in several types of tumors, including melanomas. The expression of different S100 proteins (A2, A6 and A8/A9 or A12) was examined in 42 Spitz nevi, 105 melanomas, and 73 melanocytic nevi to test the hypothesis that their expression differs among these entities and may contribute to the distinction between these entities. The results showed an up-regulation of S100A6 protein in Spitz nevi, melanomas, and melanocytic nevi but with a different percentage of positivity and pattern of immunoreactivity. The differences between these three entities were statistically significant (P <.001). All 42 Spitz nevi (100%) showed strong and diffuse S100A6 protein expression, both in junctional and in dermal components of the nevi. Thirty-three percent of melanomas expressed S100A6 (35/105). The expression was mainly weak (30/35) and patchy in the dermal component and was negative or minimal in the junctional component. Fifty-six percent of different subtypes of melanocytic nevi (41/73) expressed S100A6, almost all of them weakly (40/41) and in the dermal component. Normal intraepidermal melanocytes were negative. The melanocytic cells in these three entities did not express S100A2, S100A8/A9 or A12. However, an up regulation of S100A2 and S100A8/A9 or A12 proteins was observed in normal keratinocytes in the epidermis overlying Spitz nevi and melanomas, without differences. In summary, a simple immunohistochemical test for S100A6 protein differentiated between Spitz nevi, melanomas, and melanocytic nevi. This marker could be used when the distinction is very difficult or controversial in routine studies, especially when there is a junctional component. Further molecular analyses of the S100A6 protein and gene should be performed to study the underlying genetic bases for such differences. PMID- 12748259 TI - Pathologists and patients: can we talk? PMID- 12748260 TI - Comments and reflections on "Pathologists and patients: can we talk?". PMID- 12748258 TI - Leiomyosarcoma of urinary bladder following cyclophosphamide therapy: report of two cases. AB - Leiomyosarcoma of urinary bladder is rare, although it is the most common mesenchymal tumor in adults. We report two cases of this tumor following cyclophosphamide therapy. The first case is from a 53-year-old man with Wegener's granulomatosis treated for 6 years with cyclophosphamide. He presented with painless hematuria, and the initial biopsy of the bladder tumor revealed a malignant spindle cell neoplasm. A final diagnosis of leiomyosarcoma was made on radical cystoprostatectomy. The second example is from a 21-year-old man who had received cyclophosphamide in early infancy for a bilateral retinoblastoma. He also presented with painless hematuria, and a bladder tumor was resected transurethrally and diagnosed as leiomyosarcoma. He underwent partial cystectomy two months later. Cyclophosphamide, when used for a neoplastic or non-neoplastic condition, is associated with an increased risk of developing bladder cancer. The distribution of histologic subtypes differs from that seen in spontaneous bladder tumors. A review of the literature shows an increased proportion of squamous cell carcinomas and sarcomas, especially leiomyosarcomas in cyclophosphamide exposed patients. Acrolein, a cytotoxic metabolite of cyclophosphamide excreted in urine, is regarded as the most likely causative agent. PMID- 12748261 TI - The expanding role of common respiratory viruses in human disease. PMID- 12748262 TI - Five postulates for resolving outbreaks of infectious disease. AB - Outbreaks of infection challenge the surveillance of infectious disease, but they also offer opportunities to improve and refine it. An outbreak may be the first sign of an emerging pathogen or it may draw attention to a new risk group or route of infection. Postulates analogous to those used a century ago by Robert Koch to prove the microbial aetiology of infectious diseases can be employed to verify the existence of an outbreak, demonstrate its cause and pinpoint its origins. In doing this, high-resolution molecular finger printing of micro organisms has now assumed a crucial role. Without formal analysis based on postulates, the existence, extent and source of outbreaks may be overlooked and public health interventions misapplied or lost. PMID- 12748263 TI - Development of acquired immunity to Salmonella. AB - Salmonella enterica serovar Typhi (S. typhi) causes human typhoid fever, a serious and widespread disease in developing countries. Other Salmonella serovars are associated with food-borne infections. The recent emergence of multi-drug resistant Salmonella strains highlights the need for better preventive measures, including vaccination. The available vaccines against Salmonella infection do not confer optimal protection. The design of new Salmonella vaccines must be based on the identification of suitable virulence genes and on knowledge of the immunological mechanisms of resistance to the disease. Control and clearance of a vaccine strain rely on the phagocyte oxidative burst, reactive nitrogen intermediates, inflammatory cytokines and CD4(+) TCR-alphabeta(+) T cells and are controlled by genes including NRAMP1 and MHC class II. Vaccine-induced resistance to reinfection requires the presence of Th1-type immunological memory and anti Salmonella antibodies. The interaction between T and B cells is essential for the development of resistance following vaccination. The identification of immunodeficiencies that render individuals more susceptible to salmonellosis must be taken into consideration when designing and testing live attenuated Salmonella vaccines. An ideal live Salmonella vaccine should therefore be safe, regardless of the immunological status of the vaccinee, but still immunogenic. PMID- 12748264 TI - NapA protects Helicobacter pylori from oxidative stress damage, and its production is influenced by the ferric uptake regulator. AB - The Helicobacter pylori protein NapA has been identified as a homologue of the Escherichia coli protein Dps. It is shown in this study that, like Dps, NapA is produced maximally in stationary phase cells and contributes to the ability of H. pylori to survive under oxidative stress conditions. Moreover, NapA co-localizes with the nuclear material, suggesting that it can interact with DNA in vivo. Furthermore, it is demonstrated that repression of NapA production by iron starvation was not so pronounced in a H. pylori fur mutant, suggesting that the ferric uptake regulator (Fur) is involved in napA regulation, and a potential fur box by which this control could be mediated is identified. This finding is consistent with the regulation of iron-binding proteins by Fur and also the modulation of Fur during oxidative stress, thus allowing NapA levels to be increased in the environmental conditions under which its ability to protect DNA from attack by toxic free radicals is most beneficial to the cell. PMID- 12748265 TI - Crystallization of urine mineral components may depend on the chemical nature of Proteus endotoxin polysaccharides. AB - Formation of infectious urinary calculi is the most common complication accompanying urinary tract infections by members of the genus Proteus. The major factor involved in stone formation is the urease produced by these bacteria, which causes local supersaturation and crystallization of magnesium and calcium phosphates as carbonate apatite [Ca(10)(PO(4))(6).CO(3)] and struvite (MgNH(4)PO(4).6H(2)O), respectively. This effect may also be enhanced by bacterial polysaccharides. Macromolecules of such kind contain negatively charged residues that are able to bind Ca(2+) and Mg(2+), leading to the accumulation of these ions around bacterial cells and acceleration of the crystallization process. The levels of Ca(2+) and Mg(2+) ions bound by whole Proteus cells were measured, as well as the chemical nature of isolated LPS polysaccharides, and the intensity of the in vitro crystallization process was compared in a synthetic urine. The results suggest that the sugar composition of Proteus LPS may either enhance or inhibit the crystallization of struvite and apatite, depending on its chemical structure and ability to bind cations. This points to the increased importance of endotoxin in urinary tract infections. PMID- 12748266 TI - Biosynthesis of chondroitinase and hyaluronidase by different strains of Paracoccidioides brasiliensis. AB - The biosynthesis of chondroitinase and hyaluronidase by different isolates of Paracoccidioides brasiliensis was investigated in 20 strains isolated from patients (17 strains), a penguin (Pygocelis adeliae, one strain), an armadillo (Dasypus novemcinctus, one strain) and the environment (dog food, one strain). All the P. brasiliensis isolates studied had the ability to produce chondroitinase and hyaluronidase, although differences in colony morphology and enzyme production were detected among them. These results suggest that further investigations should be carried out in the clinical field in order to clarify the potential role of P. brasiliensis enzyme production in the pathogenesis of paracoccidioidomycosis. PMID- 12748267 TI - Lysogeny and bacteriophage host range within the Burkholderia cepacia complex. AB - The Burkholderia cepacia complex comprises a group of nine closely related species that have emerged as life-threatening pulmonary pathogens in immunocompromised patients, particularly individuals with cystic fibrosis or chronic granulomatous disease. Attempts to explain the genomic plasticity, adaptability and virulence of the complex have paid little attention to bacteriophages, particularly the potential contribution of lysogenic conversion and transduction. In this study, lysogeny was observed in 10 of 20 representative strains of the B. cepacia complex. Three temperate phages and five lytic phages isolated from soils, river sediments or the plant rhizosphere were chosen for further study. Six phages exhibited T-even morphology and two were lambda-like. The host range of individual phages, when tested against 66 strains of the B. cepacia complex and a representative panel of other pseudomonads, was not species specific within the B. cepacia complex and, in some phages, included Burkholderia gladioli and Pseudomonas aeruginosa. These new data indicate a potential role for phages of the B. cepacia complex in the evolution of these soil bacteria as pathogens of plants, humans and animals, and as novel therapeutic agents. PMID- 12748268 TI - Incidence of virulence determinants in clinical Enterococcus faecium and Enterococcus faecalis isolates collected in Sardinia (Italy). AB - Enterococci are widely distributed in the environment; within the human body, they are normal commensals of the oral cavity, gastrointestinal tract and vagina. In recent years, enterococci have become one of the most frequent causes of acquired nosocomial infections worldwide. The molecular mechanism of virulence of these bacteria is still not completely understood. The aims of this work were to characterize phenotypically 47 isolates of Enterococcus faecalis and Enterococcus faecium collected in Sardinia (Italy) by their abilities to adhere to different epithelial cell lines (Vero and Caco-2 cells) and to associate their phenotypes with the presence of known virulence genes detected within their genomes by PCR. The following genes were amplified: AS (aggregation substance), esp (surface protein gene), ace (accessory colonization factor), efaA (E. faecalis endocarditis antigen) and gelE (gelatinase). The virulence genes were detected in E. faecalis isolates only, with the exception of esp, which was found in both species. The phenotypic and genotypic results were also compared with the susceptibility of isolates to various antibiotics. PMID- 12748269 TI - Genotypic and phenotypic characterization of attaching and effacing Escherichia coli (AEEC) isolated from children with and without diarrhoea in Londrina, Brazil. AB - Attaching and effacing Escherichia coli (AEEC) have been implicated in diarrhoea in humans in several countries. A total of 919 E. coli strains, isolated from 125 children with diarrhoea and 98 without diarrhoea, was investigated by PCR for the presence of the EAF, bfp, eae and stx genes. Thirty-four of these isolates were found to carry the eae gene; they were isolated from 27 (79.4 %) children with diarrhoea and seven (20.6 %) controls, in the city of Londrina, Brazil. These strains were investigated for their genotypic and phenotypic characteristics. Different genetic profiles were observed; strains containing the eae gene alone were most common (47.1 %). The characteristic genetic profile of typical enteropathogenic E. coli (EPEC), eae, bfp and EAF, was only found in isolates from children with diarrhoea. The stx gene was not detected in any of the 34 strains studied. Ten (29.4 %) strains were negative in the fluorescent actin staining test. Localized adhesion (LA) was the most common pattern of adhesion (44.1 %), followed by the aggregative adhesion (AA) (23.5 %) and localized adhesion-like (LAL) (14.7 %) patterns. The results showed a strong association between strains presenting the LA pattern and diarrhoea. Forty-seven per cent of the strains studied belonged to classical O-serogroups of EPEC. The most common serotype found was O119 : H6; these isolates all showed the LA pattern, were positive for fluorescent actin-staining and were associated with diarrhoea. Intimin beta was detected in seven strains, four of which belonged to serotype O119 : H6 and three to serotype ONT : H7; all were associated with diarrhoea. On the other hand, intimin epsilon was detected in two strains of serotype O111 : H38 and one of serotype ONT : H19, isolated from children without diarrhoea. To our knowledge, this is the first report of the occurrence of intimin epsilon in strains of E. coli isolated from humans in Brazil. PMID- 12748270 TI - Nucleotide sequence-based typing of meningococci directly from clinical samples. AB - The unpredictable characteristics of meningococcal disease (MD) make outbreaks complicated to monitor and consequently lead to high levels of public anxiety. Traditional molecular techniques have been utilized in order to understand better the epidemiology of MD, but some have disadvantages such as being highly specialized and labour-intensive, with low reproducibility. Some of these problems have been overcome by using multilocus sequence typing (MLST). This technique exploits the unambiguous nature and electronic portability of nucleotide sequencing data for the characterization of micro-organisms. The need for enhanced surveillance of MD after the introduction of serogroup C conjugate vaccines means that it is important to gain typing information from the infecting organism in the absence of a culture isolate. Here, the application of MLST for the laboratory confirmation and characterization of Neisseria meningitidis directly from clinical samples is described. This involved using a newly designed set of primers that were complementary to nucleotide sequences external to the existing MLST primers already in use for culture-based MLST of meningococci. This combination has produced a highly sensitive procedure to allow the efficient genotypic characterization of meningococci directly from clinical samples. PMID- 12748271 TI - Evaluation of selective media for the isolation of Brachyspira aalborgi from human faeces. AB - The purposes of this study were to identify a solid medium that supports improved growth of the anaerobic intestinal spirochaete Brachyspira aalborgi, to modify this for use as a selective isolation medium and then to test the medium for its effectiveness in isolating B. aalborgi from patients' faeces. Of the media evaluated, brain heart infusion agar (BHIA) with 10 % bovine blood (BB) was the most effective base-supplement combination for growth, with colonies attaining 1.2 mm in diameter by 21 days. Incubation in an anaerobic jar (94 % H(2), 6 % CO(2)) permitted growth of larger colonies than incubation in an anaerobic chamber (80 % N(2), 10 % H(2), 10 % CO(2)). Growth was improved only slightly at 38.5 degrees C compared with 37 degrees C. Selection of B. aalborgi from artificially seeded faeces was achieved equally well on eight different solid media containing spectinomycin (400 micro g ml(-1)) alone or in combinations with polymyxin B (5 micro g ml(-1)), colistin (25 micro g ml(-1)) and rifampicin (12.5 micro g ml(-1)). By using BHIA 10 % BB with spectinomycin plus polymyxin B, B. aalborgi was isolated from one of five human faecal samples that were positive for B. aalborgi by PCR amplification. This is the first report of the isolation of B. aalborgi from human faeces. PMID- 12748272 TI - Tracing clonality of Helicobacter pylori infecting family members from analysis of DNA sequences of three housekeeping genes (ureI, atpA and ahpC), deduced amino acid sequences, and pathogenicity-associated markers (cagA and vacA). AB - Helicobacter pylori, a Gram-negative bacterium, is a causal agent of peptic ulcers and is estimated to infect the gastric mucosa of at least half of the world's population. As primary infections are acquired mainly by household contact, studies on family clusters provide a model for investigating transmission and the natural history of initial infection. Here, sequence typing exploiting genetic variation in core fragments of three key housekeeping loci (ureI, atpA and ahpC) was used to determine clonal descent amongst isolates of ten members of four families in Northern Ireland and a family with three generations in central England. Phylogenetic analysis of each locus for 73 strains of H. pylori from 11 countries indicated high background intraspecific diversity, apart from identical paired isolates from five unrelated patients and strains with identical sequence types (STs) detected in adult members of two families. In several families carrying strains with different STs, evidence of residual clonal descent was detected at one or two loci by comparison of nucleotide and amino acid sequences. Pathogenicity-associated genotypes were heterogeneous with respect to ST and amino acid type. Analysis of these three housekeeping genes provides unique evidence for precise tracing of clonal descent in isolates of H. pylori in family groups. PMID- 12748273 TI - Increasing incidence of group A streptococcal infections amongst injecting drug users in England and Wales. AB - During 2000, the UK witnessed a sudden increase in severe infections and related deaths in injecting drug users (IDUs), sparking off a UK-wide investigation. A worrying upward trend in severe group A streptococcal (GAS) infections has recently been observed in IDUs based upon isolate referrals to the PHLS Respiratory and Systemic Infection Laboratory. Most cases were young male adults who presented with skin sepsis and bacteraemia. Serotyping revealed a diverse range of M types, with higher types predominating in some geographical areas. The data suggest that GAS invasive soft-tissue infections may present in an epidemic fashion among IDUs in the absence of a common source. PMID- 12748274 TI - Molecular comparison of bacterial isolates from blood with strains colonizing pharynx and intestine in immunocompromised patients with sepsis. AB - Most causative organisms of sepsis in immunocompromised patients are the same species as those that colonize their own nasopharynx or intestinal tract. To determine whether the strains recovered from blood originate mainly from patients' own flora, isolates from blood and throat and/or stool were investigated by genomic analyses. Surveillance cultures of throat and stool were taken prospectively from cancer patients being treated with intensive chemotherapy followed by haematopoietic stem-cell transplantation. In those cases of sepsis in which the isolate from blood was the same species as that from the throat and/or stool, the genomic profiles of the isolates were compared by PFGE. Ten cases of blood culture-positive sepsis were documented in six of 14 subjects during a 2 year period; isolates of Pseudomonas aeruginosa, Staphylococcus epidermidis, Enterococcus sp., viridans streptococci and Fusobacterium sp. were recovered from blood. In five of seven cases in which the blood isolate was the same species as that from the throat or stool, the genotypes of the isolates from both sites were identical. In the majority of immunocompromised patients, the causative organisms of bloodstream infections originated mainly from their own flora. PMID- 12748275 TI - Role of elastase in a mouse model of chronic respiratory Pseudomonas aeruginosa infection that mimics diffuse panbronchiolitis. AB - Pseudomonas aeruginosa frequently colonizes the respiratory tract of patients suffering from cystic fibrosis (CF) and diffuse panbronchiolitis (DPB). However, the relationship between lung inflammation and extracellular products of P. aeruginosa is not well-defined. To assess the role of elastase released by P. aeruginosa in DPB, a murine model of DPB was employed in this study. Mice were inoculated with either P. aeruginosa PAO1 or PAO-E64; the latter produces elastase with greatly reduced enzymic activity. Throughout the 90-day experiments, counts of viable bacteria from the PAO1- and PAO-E64-infected mice were found to be equivalent. However, the number of lymphocytes isolated from the lungs of PAO-E64-infected mice was significantly lower than the number isolated from the lungs of PAO1-infected animals. Histopathological examination of the lungs of mice infected by PAO1 on day 90 revealed an intense accumulation of chronic respiratory cells surrounding the bronchi, in sharp contrast to the more localized inflammatory response found in those mice infected by PAO-E64. These data suggest that P. aeruginosa elastase (PE) is a potent inflammatory factor in a mouse model of DPB and that the control of PE release by P. aeruginosa may be beneficial for patients with DPB. PMID- 12748276 TI - Identification of E-box factor TFE3 as a functional partner for the E2F3 transcription factor. AB - Various studies have demonstrated a role for E2F proteins in the control of transcription of genes involved in DNA replication, cell cycle progression, and cell fate determination. Although it is clear that the functions of the E2F proteins overlap, there is also evidence for specific roles for individual E2F proteins in the control of apoptosis and cell proliferation. Investigating protein interactions that might provide a mechanistic basis for the specificity of E2F function, we identified the E-box binding factor TFE3 as an E2F3-specific partner. We also show that this interaction is dependent on the marked box domain of E2F3. We provide evidence for a role for TFE3 in the synergistic activation of the p68 subunit gene of DNA polymerase alpha together with E2F3, again dependent on the E2F3 marked box domain. Chromatin immunoprecipitation assays showed that TFE3 and E2F3 were bound to the p68 promoter in vivo and that the interaction of either E2F3 or TFE3 with the promoter was facilitated by the presence of both proteins. In contrast, neither E2F1 nor E2F2 interacted with the p68 promoter under these conditions. We propose that the physical interaction of TFE3 and E2F3 facilitates transcriptional activation of the p68 gene and provides strong evidence for the specificity of E2F function. PMID- 12748277 TI - The Rad51 pathway of telomerase-independent maintenance of telomeres can amplify TG1-3 sequences in yku and cdc13 mutants of Saccharomyces cerevisiae. AB - In the yeast Saccharomyces cerevisiae, Cdc13, Yku, and telomerase define three parallel pathways for telomere end protection that prevent chromosome instability and death by senescence. We report here that cdc13-1 yku70delta mutants generated telomere deprotection-resistant cells that, in contrast with telomerase-negative senescent cells, did not display classical crisis events. cdc13-1 yku70delta cells survived telomere deprotection by exclusively amplifying TG(1-3) repeats (type II recombination). In a background lacking telomerase (tlc1delta), this process predominated over type I recombination (amplification of subtelomeric Y' sequences). Strikingly, inactivation of the Rad50/Rad59 pathway (which is normally required for type II recombination) in cdc13-1 yku70delta or yku70delta tlc1delta mutants, but also in cdc13-1 YKU70(+) tlc1delta mutants, still permitted type II recombination, but this process was now entirely dependent on the Rad51 pathway. In addition, delayed senescence was observed in cdc13-1 yku70delta rad51delta and cdc13-1 tlc1delta rad51delta cells. These results demonstrate that in wild-type cells, masking by Cdc13 and Yku prevents the Rad51 pathway from amplifying telomeric TG(1-3) sequences. They also suggest that Rad51 is more efficient than Rad50 in amplifying the sequences left uncovered by the absence of Cdc13 or Yku70. PMID- 12748278 TI - p27Kip1 inhibition of GRB2-SOS formation can regulate Ras activation. AB - p27(Kip1) (p27) is often inappropriately downregulated in aggressive human cancers. Although p27 can inhibit cyclin-dependent kinases (CDKs), low p27 does not always correlate with increased CDK activity. Furthermore, cells derived from p27(-/-) mice respond to antimitogens, maintain restriction point control, and do not deregulate CDKs. Thus, disruption of a p27 function other than CDK inhibition may contribute to the disease state. A yeast two-hybrid screen identified growth factor receptor-bound protein 2 (GRB2) as a p27 binding partner. We now demonstrate that p27 can inhibit GRB2 function by blocking its association with the guanine nucleotide exchange factor SOS. Endogenous p27 is rapidly exported from the nucleus to the cytoplasm in response to mitogen stimulation, where it binds GRB2 concomitant with a decrease in GRB2-associated SOS. As predicted, mitogen-stimulated p27(-/-) cells maintained their GRB2-SOS complexes for significantly longer. The Ras/mitogen-activated protein kinase pathway does not appear to be deregulated in cells lacking p27 despite excess GRB2-SOS, suggesting that additional control mechanisms are present. A transient-transfection approach was employed to show that p27 can inhibit Ras activation by targeting GRB2 and further revealed that the CDK and GRB2 inhibitory functions of p27 are separable and distinct. Thus, p27 downregulation may compromise control of Ras, one of the most common oncogenic events in human cancer. PMID- 12748279 TI - Protein tyrosine phosphatase 1B attenuates growth hormone-mediated JAK2-STAT signaling. AB - Protein tyrosine phosphatase-1B (PTP-1B) attenuates insulin, PDGF, EGF, and IGF-I signaling by dephosphorylating tyrosine residues located in the tyrosine kinase domain of the corresponding receptors. More recently, PTP-1B was shown to modulate the action of cytokine signaling via the nonreceptor tyrosine kinase JAK2. Transmission of the growth hormone (GH) signal also depends on JAK2, raising the possibility that PTP-1B modulates GH action. Consistent with this hypothesis, GH increased the abundance of tyrosine-phosphorylated JAK2 associated with a catalytically inactive mutant of PTP-1B. GH-induced JAK2 phosphorylation was greater in knockout (KO) than in wild-type (WT) PTP-1B embryonic fibroblasts and resulted in increased tyrosine phosphorylation of STAT3 and STAT5, while overexpression of PTP-1B reduced the GH-mediated activation of the acid-labile subunit gene. To evaluate the in vivo relevance of these observations, mice were injected with GH under fed and fasted conditions. As expected, tyrosine phosphorylation of JAK2 and STAT5 occurred readily in the livers of fed WT mice and was almost completely abolished during fasting. In contrast, resistance to the action of GH was severely impaired in the livers of fasted KO mice. These results indicate that PTP-1B regulates GH signaling by reducing the extent of JAK2 phosphorylation and suggest that PTP-1B is essential for limiting the action of GH during metabolic stress such as fasting. PMID- 12748280 TI - Progesterone and glucocorticoid receptors recruit distinct coactivator complexes and promote distinct patterns of local chromatin modification. AB - It is well established that steroid receptor function requires interaction with coactivators. However, the mechanisms through which steroid receptors elicit precise assembly of coactivator complexes and the way the steroid activation signal is transduced remain elusive. Using a T47D cell line stably integrated with a mouse mammary tumor virus-chloramphenicol acetyltransferase (MMTV-CAT) reporter, we demonstrate that specific steroid receptors exhibit preferential recruitment of SRC-1 family coactivators, which determines the subsequent recruitment of specific downstream coregulator molecules. Upon ligand treatment, progesterone receptor (PR) interacted preferentially with SRC-1, which recruited CBP and significantly enhanced acetylation at K5 of histone H4. In contrast, activated glucocorticoid receptor (GR) preferentially associated with SRC-2 (TIF 2/GRIP-1), which subsequently recruited pCAF and led to specific modification of histone H3, suggesting that specific coactivators recruit distinct histone acetyltransferases to modulate the transcription of steroid-responsive genes. Loss-of-function experiments further support the predicted roles of SRC-1 and SRC 2 in, respectively, PR- and GR-mediated transcription on the MMTV promoter. This study indicates that differential recruitment of coactivators by nuclear receptors determines the assembly of coactivator complexes on target promoters to mediate specific transcription signals. PMID- 12748281 TI - Regulation of BRCA1 transcription by specific single-stranded DNA binding factors. AB - Since the majority of high-grade breast cancers express reduced levels of BRCA1 mRNA, we investigated the factors regulating BRCA1 transcription. Factors with specific affinity for the previously identified positive regulatory region (PRR) in the BRCA1 promoter were purified from whole-cell extracts. Identified proteins included replication protein A and a series of related factors with affinity for the sense strand of PRR. A subset of the identified factors activated the BRCA1 promoter. Identification of these families of proteins regulating the BRCA1 promoter represents an important step in the comprehension of the mechanisms responsible for breast cancer development. PMID- 12748282 TI - Polyphosphate loss promotes SNF/SWI- and Gcn5-dependent mitotic induction of PHO5. AB - Approximately 800 transcripts in Saccharomyces cerevisiae are cell cycle regulated. The oscillation of approximately 40% of these genes, including a prominent subclass involved in nutrient acquisition, is not understood. To address this problem, we focus on the mitosis-specific activation of the phosphate-responsive promoter, PHO5. We show that the unexpected mitotic induction of the PHO5 acid phosphatase in rich medium requires the transcriptional activators Pho4 and Pho2, the cyclin-dependent kinase inhibitor Pho81, and the chromatin-associated enzymes Gcn5 and Snf2/Swi2. PHO5 mitotic activation is repressed by addition of orthophosphate, which significantly increases cellular polyphosphate. Polyphosphate levels also fluctuate inversely with PHO5 mRNA during the cell cycle, further substantiating an antagonistic link between this phosphate polymer and PHO5 mitotic regulation. Moreover, deletion of PHM3, required for polyphosphate accumulation, leads to premature onset of PHO5 expression, as well as an increased rate, magnitude, and duration of PHO5 activation. Orthophosphate addition, however, represses mitotic PHO5 expression in a phm3delta strain. Thus, polyphosphate per se is not necessary to repress PHO transcription but, when present, replenishes cellular phosphate during nutrient depletion. These results demonstrate a dynamic mechanism of mitotic transcriptional regulation that operates mostly independently of factors that drive progression through the cell cycle. PMID- 12748283 TI - Tristetraprolin and its family members can promote the cell-free deadenylation of AU-rich element-containing mRNAs by poly(A) ribonuclease. AB - Eukaryotic mRNA stability can be influenced by AU-rich elements (AREs) within mRNA primary sequences. Tristetraprolin (TTP) is a CCCH tandem zinc finger protein that binds to ARE-containing transcripts and destabilizes them, apparently by first promoting the removal of their poly(A) tails. We developed a cell-free system in which TTP and its related proteins stimulated the deadenylation of ARE-containing, polyadenylated transcripts. Transcript deadenylation was not stimulated when a mutant TTP protein was used that was incapable of RNA binding, nor when a mutant ARE was present that did not bind TTP. The ability of TTP to promote transcript deadenylation required Mg(2+), but not ATP or prior capping of the RNA substrate. Cotransfection and additivity studies with the poly(A) RNase (PARN) demonstrated that TTP promoted the ability of this enzyme to deadenylate ARE-containing, polyadenylated transcripts, while having no effect on transcripts lacking an ARE. There was no effect of TTP to act synergistically with enzymatically inactive PARN mutants. We conclude that TTP can promote the deadenylation of ARE-containing, polyadenylated substrates by PARN. This interaction may be responsible for the ability of TTP and its family members to promote the deadenylation of such transcripts in intact cells. PMID- 12748284 TI - Inactivation of dual-specificity phosphatases is involved in the regulation of extracellular signal-regulated kinases by heat shock and hsp72. AB - Extracellular signal-regulated kinase 1 (ERK1) and ERK2 (ERK1/2) dramatically enhance survival of cells exposed to heat shock. Using Cos-7 cells and primary human fibroblasts (IMR90 cells), we demonstrated that heat shock activates ERKs via two distinct mechanisms: stimulation of the ERK-activating kinases, MEK1/2, and inhibition of ERK dephosphorylation. Under milder heat shock conditions, activation of ERKs proceeded mainly through stimulation of MEK1/2, whereas under more severe heat shock MEK1/2 could no longer be activated and the inhibition of ERK phosphatases became critical. In Cos-7 cells, nontoxic heat shock caused rapid inactivation of the major ERK phosphatase, MKP-3, by promoting its aggregation, so that in cells exposed to 45 degrees C for 20 min, 90% of MKP-3 became insoluble. MKP-3 aggregation was reversible and, 1 h after heat shock, MKP 3 partially resolubilized. The redistribution of MKP-3 correlated with an increased rate of ERK dephosphorylation. Similar heat-induced aggregation, followed by partial resolubilization, was found with a distinct dual-specificity phosphatase MKP-1 but not with MKP-2. Therefore, MKP-3 and MKP-1 appeared to be critical heat-labile phosphatases involved in the activation of ERKs by heat shock. Expression of the major heat shock protein Hsp72 inhibited activation of MEK1/2 and prevented inactivation of MKP-3 and MKP-1. Hsp72DeltaEEVD mutant lacking a chaperone activity was unable to protect MKP-3 from heat inactivation but interfered with MEK1/2 activation similar to normal Hsp72. Hence, Hsp72 suppressed ERK activation by both protecting dual-specificity phosphatases, which was dependent on the chaperone activity, and suppressing MEK1/2, which was independent of the chaperone activity. PMID- 12748285 TI - Transcription of endogenous and exogenous R2 elements in the rRNA gene locus of Drosophila melanogaster. AB - R2 retrotransposons insert into the rRNA-encoding units (rDNA units) that form the nucleoli of insects. We have utilized an R2 integration system in Drosophila melanogaster to study transcription of foreign sequences integrated into the R2 target site of the 28S rRNA genes. The exogenous sequences were cotranscribed at dramatically different levels which closely paralleled the level of transcription of the endogenous R1 and R2 elements. Transcription levels were inversely correlated with the number of uninserted rDNA units, variation in this number having been brought about by the R2 integration system itself. Females with as few as 20 uninserted rDNA units per X chromosome had expression levels of endogenous and exogenous insertion sequences that were 2 orders of magnitude higher than lines that contained over 80 uninserted rDNA units per chromosome. R2 insertions only 167 bp in length exhibited this range of transcriptional regulation. Analysis of transcript levels in males suggested R2 insertions on the Y chromosome are not down-regulated to the same extent as insertions on the X chromosome. These results suggest that transcription of the rDNA units can be tightly regulated, but this regulation gradually breaks down as the cell approaches the minimum number of uninserted genes needed for survival. PMID- 12748286 TI - Inhibition of p300/CBP by early B-cell factor. AB - Early B-cell factor (EBF) is a DNA binding protein required for early B-cell development. It activates transcription of several B-cell-specific genes, including the lambda5 gene, which encodes a protein necessary for signaling by the pre-B-cell receptor. In an effort to understand the mechanism by which EBF activates transcription, we examined its interaction with the coactivator protein p300/CBP. We found that two domains of EBF each bind the histone acetyltransferase (HAT)/CH3 domain of p300/CBP both in vitro and in vivo. Surprisingly, transcriptional activation by EBF was not sensitive to E1A, a potent p300/CBP inhibitor. In fact, overexpressed EBF mimicked E1A by severely repressing the activity of several other transcription factors, including E47, a protein that acts cooperatively with EBF to promote transcription of the lambda5 gene. This broad inhibitory profile correlated with EBF's ability to repress the HAT activity of p300/CBP in vivo and in vitro. However, such a repressed complex is not likely to form at the lambda5 promoter in vivo since (i) EBF could not bind p300/CBP and DNA simultaneously and (ii) the cooperativity imparted by E47 was sensitive to E1A. Our data reveal an intriguing inhibitory property of EBF-a property shared only by E1A, Twist, Pu.1, and the Hox family of homeodomain proteins-and suggest that E47 and EBF play distinct roles during lambda5 promoter activation. PMID- 12748287 TI - Identification of tetratricopeptide repeat 1 as an adaptor protein that interacts with heterotrimeric G proteins and the small GTPase Ras. AB - The biological functions of heterotrimeric G proteins and small GTPases are modulated by both extracellular stimuli and intracellular regulatory proteins. Using Saccharomyces cerevisiae two-hybrid screening, we identified tetratricopeptide repeat 1 (TPR1), a 292-amino-acid protein with three TPR motifs, as a Galpha16-binding protein. The interaction was confirmed both in vitro and in transfected mammalian cells, where TPR1 also binds to several other Galpha proteins. TPR1 was found to interact with Ha-Ras preferentially in its active form. Overexpression of TPR1 promotes accumulation of active Ras. TPR1 was found to compete with the Ras-binding domain (RBD) of Raf-1 for binding to the active Ras, suggesting that it may also compete with Ras GTPase-activating protein, thus contributing to the accumulation of GTP-bound Ras. Expression of Galpha16 strongly enhances the interaction between TPR1 and Ras. Removal of the TPR1 N-terminal 112 residues abolishes potentiation by Galpha16 while maintaining the interaction with Galpha16 and the ability to discriminate active Ras from wild-type Ras. We have also observed that LGN, a Galphai-interacting protein with seven TPR motifs, binds Ha-Ras. Thus, TPR1 is a novel adaptor protein for Ras and selected Galpha proteins that may be involved in protein-protein interaction relating to G-protein signaling. PMID- 12748290 TI - Two distinct phosphorylation pathways have additive effects on Abl family kinase activation. AB - The activities of the related Abl and Arg nonreceptor tyrosine kinases are kept under tight control in cells, but exposure to several different stimuli results in a two- to fivefold stimulation of kinase activity. Following the breakdown of inhibitory intramolecular interactions, Abl activation requires phosphorylation on several tyrosine residues, including a tyrosine in its activation loop. These activating phosphorylations have been proposed to occur either through autophosphorylation by Abl in trans or through phosphorylation of Abl by the Src nonreceptor tyrosine kinase. We show here that these two pathways mediate phosphorylation at distinct sites in Abl and Arg and have additive effects on Abl and Arg kinase activation. Abl and Arg autophosphorylate at several sites outside the activation loop, leading to 5.2- and 6.2-fold increases in kinase activity, respectively. We also find that the Src family kinase Hck phosphorylates the Abl and Arg activation loops, leading to an additional twofold stimulation of kinase activity. The autoactivation pathway may allow Abl family kinases to integrate or amplify cues relayed by Src family kinases from cell surface receptors. PMID- 12748288 TI - Loss of oncogenic H-ras-induced cell cycle arrest and p38 mitogen-activated protein kinase activation by disruption of Gadd45a. AB - The activation of p53 is a guardian mechanism to protect primary cells from malignant transformation; however, the details of the activation of p53 by oncogenic stress are still incomplete. In this report we show that in Gadd45a(-/ ) mouse embryo fibroblasts (MEF), overexpression of H-ras activates extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) but not p38 kinase, and this correlates with the loss of H-ras-induced cell cycle arrest (premature senescence). Inhibition of p38 mitogen-activated protein kinase (MAPK) activation correlated with the deregulation of p53 activation, and both a p38 MAPK chemical inhibitor and the expression of a dominant-negative p38alpha inhibited p53 activation in the presence of H-ras in wild-type MEF. p38, but not ERK or JNK, was found in a complex with Gadd45 proteins. The region of interaction was mapped to amino acids 71 to 96, and the central portion (amino acids 71 to 124) of Gadd45a was required for p38 MAPK activation in the presence of H-ras. Our results indicate that this Gadd45/p38 pathway plays an important role in preventing oncogene-induced growth at least in part by regulating the p53 tumor suppressor. PMID- 12748289 TI - Reduced proliferative capacity of hematopoietic stem cells deficient in Hoxb3 and Hoxb4. AB - Several homeobox transcription factors, such as HOXB3 and HOXB4, have been implicated in regulation of hematopoiesis. In support of this, studies show that overexpression of HOXB4 strongly enhances hematopoietic stem cell regeneration. Here we find that mice deficient in both Hoxb3 and Hoxb4 have defects in endogenous hematopoiesis with reduced cellularity in hematopoietic organs and diminished number of hematopoietic progenitors without perturbing lineage commitment. Analysis of embryonic day 14.5 fetal livers revealed a significant reduction in the hematopoietic stem cell pool, suggesting that the reduction in cellularity observed postnatally is due to insufficient expansion during fetal development. Primitive Lin(-) ScaI(+) c-kit(+) hematopoietic progenitors lacking Hoxb3 and Hoxb4 displayed impaired proliferative capacity in vitro. Similarly, in vivo repopulating studies of Hoxb3/Hoxb4-deficient hematopoietic cells resulted in lower repopulating capability compared to normal littermates. Since no defects in homing were observed, these results suggest a slower regeneration of mutant HSC. Furthermore, treatment with cytostatic drugs demonstrated slower cell cycle kinetics of hematopoietic stem cells deficient in Hoxb3 and Hoxb4, resulting in increased tolerance to antimitotic drugs. Collectively, these data suggest a direct physiological role of Hoxb4 and Hoxb3 in regulating stem cell regeneration and that these genes are required for maximal proliferative response. PMID- 12748291 TI - PIM3 proto-oncogene kinase is a common transcriptional target of divergent EWS/ETS oncoproteins. AB - Despite significant structural diversity, present evidence suggests that EWS/ETS fusion proteins promote oncogenesis by transcriptionally modulating a common set of target genes. In order to identify these genes, microarray expression analyses were performed on NIH 3T3 polyclonal populations expressing one of three EWS/ETS fusion genes. The majority of these genes can be grouped into seven functional categories, including cellular metabolism and signal transduction. The biologic significance of these target genes was pursued. The effects of modulating genes involved in metabolism were assessed by flux studies and demonstrated shifts in glucose utilization and lactate production as a result of EWS/FLI1 expression. The proto-oncogene coding for serine/threonine kinase PIM3 was found to one of several genes encoding signal transduction proteins that were up-regulated by EWS/ETS fusions. PIM3 was found to be expressed in a panel of human Ewing's family tumor cell lines. Forced expression of PIM3 promoted anchorage-independent growth. Coexpression of a kinase-deficient PIM3 mutant attenuated EWS/FLI1 mediated NIH 3T3 tumorigenesis in immunodeficent mice. PMID- 12748292 TI - Yeast Pak1 kinase associates with and activates Snf1. AB - Members of the Snf1/AMP-activated protein kinase family are activated under conditions of nutrient stress by a distinct upstream kinase. Here we present evidence that the yeast Pak1 kinase functions as a Snf1-activating kinase. Pak1 associates with the Snf1 kinase in vivo, and the association is greatly enhanced under glucose-limiting conditions when Snf1 is active. Snf1 kinase complexes isolated from pak1Delta mutant strains show reduced specific activity in vitro, and affinity-purified Pak1 kinase is able to activate the Snf1-dependent phosphorylation of Mig1 in vitro. Purified Pak1 kinase promotes the phosphorylation of the Snf1 polypeptide on threonine 210 within the activation loop in vitro, and an increased dosage of the PAK1 gene causes increased Snf1 threonine 210 phosphorylation in vivo. Deletion of the PAK1 gene does not produce a Snf phenotype, suggesting that one or more additional protein kinases is able to activate Snf1 in vivo. However, deletion of the PAK1 gene suppresses many of the phenotypes associated with the deletion of the REG1 gene, providing genetic evidence that Pak1 activates Snf1 in vivo. The closest mammalian homologue of yeast Pak1 kinase, calcium-calmodulin-dependent protein kinase kinase beta, may play a similar role in mammalian nutrient stress signaling. PMID- 12748294 TI - The histidine triad protein Hint is not required for murine development or Cdk7 function. AB - The histidine triad (HIT) protein Hint has been found to associate with mammalian Cdk7, as well as to interact both physically and genetically with the budding yeast Cdk7 homologue Kin28. To study the function of Hint and to explore its possible role in modulating Cdk7 activity in vivo, we have characterized the expression pattern of murine Hint and generated Hint-deficient (Hint(-/-)) mice. Hint was widely expressed during mouse development, with pronounced expression in several neuronal ganglia, epithelia, hearts, and testes from embryonic day 15 onward. Despite this widespread expression, disruption of Hint did not impair murine development. Moreover, Hint-deficient mice had a normal life span and were apparently healthy. Histological examination of tissues with high Hint expression in wild-type animals did not show signs of abnormal pathology in Hint(-/-) mice. Functional redundancy within the HIT family was addressed by crossing Hint(-/-) mice with mice lacking the related HIT protein, Fhit, and by assaying the expression levels of the HIT protein gene family members Hint2 and Hint3 in Hint(+/+) and Hint(-/-) tissues. Finally, Cdk7 kinase activity and cell cycle kinetics were found to be comparable in wild-type and Hint(-/-) mouse embryonic fibroblasts, suggesting that Hint may not be a key regulator of Cdk7 activity. PMID- 12748293 TI - Interleukin-13 induction of 15-lipoxygenase gene expression requires p38 mitogen activated protein kinase-mediated serine 727 phosphorylation of Stat1 and Stat3. AB - Interleukin-13 (IL-13) is a cytokine secreted by Th2 lymphocytes that is capable of inducing expression of 15-lipoxygenase (15-LO) in primary human monocytes. We recently demonstrated that induction of 15-LO requires the activation of Jak2 and Tyk2 kinases and Stats 1, 3, 5, and 6. Since IL-13-induced 15-LO expression was inhibited by H7 (a serine-threonine kinase inhibitor), we predicted that Stat serine phosphorylation may also be crucial for 15-LO expression. In this study, we present evidence indicating that IL-13-induced 15-LO mRNA expression was detectable as early as 1 h by real-time reverse transcription-PCR. We found that IL-13 induced a time-dependent serine phosphorylation of both Stat1 and Stat3, detectable at 15 min after IL-13 treatment. In addition, the activation of p38 mitogen-activated protein kinase (MAPK) was detected in a time-dependent fashion, with peak phosphorylation at 15 min after IL-13 treatment. SB202190, a p38 MAPK specific inhibitor, markedly inhibited IL-13-induced Stat1 and Stat3 serine phosphorylation as well as DNA binding. Furthermore, treatment of cells with Stat1 or Stat3 decoys significantly impaired IL-13-induced 15-LO expression. Taken together, our results provide the first evidence that IL-13 induces p38 MAPK phosphorylation/activation, which regulates Stat1 and Stat3 serine 727 phosphorylation. Both of these events are important steps in IL-13-induced 15-LO expression in human monocytes. PMID- 12748295 TI - A functional screen in human cells identifies UBF2 as an RNA polymerase II transcription factor that enhances the beta-catenin signaling pathway. AB - beta-Catenin signaling plays an important role in the development of many organisms and has a key part in driving the malignant transformation of epithelial cells comprising a variety of cancers. beta-Catenin can activate gene expression through its association with transcription factors of the lymphoid enhancer factor 1 (LEF-1)/T-cell factor (TCF) family. We designed a screen in human cells to identify novel genes that activate a beta-catenin-LEF/TCF responsive promoter and isolated the high-mobility group box transcription factor, UBF2. UBF1 and UBF2 are splice variants of a common precursor RNA. Although UBF1 has been shown to activate RNA polymerase I-regulated genes, the function of UBF2 has remained obscure. Here, we show for the first time that both UBF1 and UBF2 activate RNA polymerase II-regulated promoters. UBF2 associates with LEF-1, as shown by coimmunoprecipitation experiments, and potentiates transcriptional activation stimulated by LEF-1/beta-catenin from a synthetic promoter with multimerized LEF/TCF binding sites and a natural cyclin D1 promoter with consensus LEF/TCF binding sites. Downregulation of endogenous UBF expression using an RNA interference approach reduces transcriptional activation of a beta catenin-LEF/TCF-responsive promoter by means of overexpressed beta-catenin, further implicating UBF as a transcriptional enhancer of the beta-catenin pathway. PMID- 12748297 TI - Cohesins determine the attachment manner of kinetochores to spindle microtubules at meiosis I in fission yeast. AB - During mitosis, sister kinetochores attach to microtubules that extend to opposite spindle poles (bipolar attachment) and pull the chromatids apart at anaphase (equational segregation). A multisubunit complex called cohesin, including Rad21/Scc1, plays a crucial role in sister chromatid cohesion and equational segregation at mitosis. Meiosis I differs from mitosis in having a reductional pattern of chromosome segregation, in which sister kinetochores are attached to the same spindle (monopolar attachment). During meiosis, Rad21/Scc1 is largely replaced by its meiotic counterpart, Rec8. If Rec8 is inactivated in fission yeast, meiosis I is shifted from reductional to equational division. However, the reason rec8Delta cells undergo equational rather than random division has not been clarified; therefore, it has been unclear whether equational segregation is due to a loss of cohesin in general or to a loss of a specific requirement for Rec8. We report here that the equational segregation at meiosis I depends on substitutive Rad21, which relocates to the centromeres if Rec8 is absent. Moreover, we demonstrate that even if sufficient amounts of Rad21 are transferred to the centromeres at meiosis I, thereby establishing cohesion at the centromeres, rec8Delta cells never recover monopolar attachment but instead secure bipolar attachment. Thus, Rec8 and Rad21 define monopolar and bipolar attachment, respectively, at meiosis I. We conclude that cohesin is a crucial determinant of the attachment manner of kinetochores to the spindle microtubules at meiosis I in fission yeast. PMID- 12748296 TI - Activin induces x-zone apoptosis that inhibits luteinizing hormone-dependent adrenocortical tumor formation in inhibin-deficient mice. AB - Inhibin and activin are members of the transforming growth factor beta (TGF-beta) family of ligands produced and secreted primarily by the gonads and adrenals. Inhibin-null (INH(-/-)) mice develop gonadal tumors and-when gonadectomized adrenocortical carcinoma. The mechanisms leading to adrenal tumorigenesis have been proposed to involve the lack of a gonadal factor and/or a compensatory increase in gonadotropins. In order to achieve elevation of gonadotropins without the concomitant loss of a gonadal hormone, we crossed INH(-/-) mice with a transgenic mouse strain that has chronically elevated luteinizing hormone (LH) levels (LH-CTP). Compound INH(-/-)-LH-CTP mice die within 6 weeks of age from severe cancer cachexia induced by large, activin-secreting ovarian tumors. Unexpectedly, INH(-/-)-LH-CTP mice not only fail to develop adrenal tumors but have smaller adrenals, with a regressed x zone, indicating that elevated LH levels are not sufficient to induce adrenal tumor formation. However, following gonadectomy, INH(-/-)-LH-CTP mice develop large, sex steroid-producing adrenal tumors that arise from the x zone, indicating a growth-promoting effect of high levels of LH on the adrenal cortex in the absence of ovarian tumors. In addition, in vivo and in vitro data indicate that activin induces apoptosis specifically in the adrenal x zone. The restricted expression of activin receptor subunits and Smad2 in cells of the adrenal x zone, together with the elevated activin levels in INH(-/-)-LH-CTP mice, supports the conclusion that activin inhibits adrenal tumor growth by inducing x-zone regression. PMID- 12748298 TI - Spatial and temporal cellular responses to single-strand breaks in human cells. AB - DNA single-strand breaks (SSB) are one of the most frequent DNA lesions produced by reactive oxygen species and during DNA metabolism, but the analysis of cellular responses to SSB remains difficult due to the lack of an experimental method to produce SSB alone in cells. By using human cells expressing a foreign UV damage endonuclease (UVDE) and irradiating the cells with UV through tiny pores in membrane filters, we created SSB in restricted areas in the nucleus by the immediate action of UVDE on UV-induced DNA lesions. Cellular responses to the SSB were characterized by using antibodies and fluorescence microscopy. Upon UV irradiation, poly(ADP-ribose) synthesis occurred immediately in the irradiated area. Simultaneously, but dependent on poly(ADP-ribosyl)ation, XRCC1 was translocated from throughout the nucleus, including nucleoli, to the SSB. The BRCT1 domain of XRCC1 protein was indispensable for its poly(ADP-ribose) dependent recruitment to the SSB. Proliferating cell nuclear antigen and the p150 subunit of chromatin assembly factor 1 also accumulated at the SSB in a detergent resistant form, which was significantly reduced by inhibition of poly(ADP-ribose) synthesis. Our results show the importance of poly(ADP-ribosyl)ation in sequential cellular responses to SSB. PMID- 12748299 TI - Hybrid embryonic stem cell-derived tetraploid mice show apparently normal morphological, physiological, and neurological characteristics. AB - ES cell-tetraploid (ES) mice are completely derived from embryonic stem cells and can be obtained at high efficiency upon injection of hybrid ES cells into tetraploid blastocysts. This method allows the immediate generation of targeted mouse mutants from genetically modified ES cell clones, in contrast to the standard protocol, which involves the production of chimeras and several breeding steps. To provide a baseline for the analysis of ES mouse mutants, we performed a phenotypic characterization of wild-type B6129S6F(1) ES mice in relation to controls of the same age, sex, and genotype raised from normal matings. The comparison of 90 morphological, physiological, and behavioral parameters revealed elevated body weight and hematocrit as the only major difference of ES mice, which exhibited an otherwise normal phenotype. We further demonstrate that ES mouse mutants can be produced from mutant hybrid ES cells and analyzed within a period of only 4 months. Thus, ES mouse technology is a valid research tool for rapidly elucidating gene function in vivo. PMID- 12748300 TI - CtBP-independent repression in the Drosophila embryo. AB - There are three mechanisms of transcriptional repression in eukaryotes. The first is quenching, whereby repressors and activators co-occupy closely linked sites and then the repressor inhibits adjacent activators. The second is direct repression, in which repressors block the function of the core transcription complex. The third is competition, in which repressors compete with activators for a common DNA-binding site. Previous studies have shown that the Drosophila melanogaster CtBP corepressor (dCtBP) is essential for the quenching activity of three short-range sequence-specific repressors in the early Drosophila embryo: Kruppel, Knirps, and Snail. Here we demonstrate that dCtBP is dispensable for target enhancers that contain overlapping activator and repressor binding sites. However, it is essential when Kruppel and Knirps repressor sites do not overlap activator sites but are instead located adjacent to either activators or the core promoter. These findings provide evidence that competition is distinct from quenching and direct repression. Quenching and direct repression depend on dCtBP, whereas competition does not. PMID- 12748302 TI - Evolutionary divergence of platelet-derived growth factor alpha receptor signaling mechanisms. AB - Receptor tyrosine kinases (RTKs) direct diverse cellular and developmental responses by stimulating a relatively small number of overlapping signaling pathways. Specificity may be determined by RTK expression patterns or by differential activation of individual signaling pathways. To address this issue we generated knock-in mice in which the extracellular domain of the mouse platelet-derived growth factor alpha receptor (PDGFalphaR) is fused to the cytosolic domain of Drosophila Torso (alpha(Tor)) or the mouse fibroblast growth factor receptor 1 (alpha(FR)). alpha(Tor) homozygous embryos exhibit significant rescue of neural crest and angiogenesis defects normally found in PDGFalphaR-null embryos yet fail to rescue skeletal or extraembryonic defects. This phenotype was associated with the ability of alpha(Tor) to stimulate the mitogen-activated protein (MAP) kinase pathway to near wild-type levels but failure to completely activate other pathways, such as phosphatidylinositol (PI) 3-kinase. The alpha(FR) chimeric receptor fails to rescue any aspect of the PDGFalphaR-null phenotype. Instead, alpha(FR) expression leads to a gain-of-function phenotype highlighted by ectopic bone development. The alpha(FR) phenotype was associated with a failure to limit MAP kinase signaling and to engage significant PI3-kinase response. These results suggest that precise regulation of divergent downstream signaling pathways is critical for specification of RTK function. PMID- 12748301 TI - In vitro import of a nuclearly encoded tRNA into mitochondria of Solanum tuberosum. AB - Some of the mitochondrial tRNAs of higher plants are nuclearly encoded and imported into mitochondria. The import of tRNAs encoded in the nucleus has been shown to be essential for proper protein translation within mitochondria of a variety of organisms. Here, we report the development of an in vitro assay for import of nuclearly encoded tRNAs into plant mitochondria. This in vitro system utilizes isolated mitochondria from Solanum tuberosum and synthetic tRNAs transcribed from cloned nuclear tRNA genes. Although incubation of radioactively labeled in vitro-transcribed tRNA(Ala), tRNA(Phe), and tRNA(Met-e) with isolated potato mitochondria resulted in importation, as measured by nuclease protection, the amount of tRNA transcripts protected at saturation was at least five times higher for tRNA(Ala) than for the two other tRNAs. This difference in in vitro saturation levels of import is consistent with the in vivo localization of these tRNAs, since cytosolic tRNA(Ala) is naturally imported into potato mitochondria whereas tRNA(Phe) and tRNA(Met-e) are not. Characterization of in vitro tRNA import requirements indicates that mitochondrial tRNA import proceeds in the absence of any added cytosolic protein fraction, involves at least one protein component on the surface of mitochondria, and requires ATP-dependent step(s) and a membrane potential. PMID- 12748304 TI - The effects of vinflunine, vinorelbine, and vinblastine on centromere dynamics. AB - Vinflunine is a novel fluorinated Vinca alkaloid currently in Phase II clinical trials, which in preclinical studies exhibited superior antitumor activity to that of two clinically useful Vinca alkaloids, vinorelbine and vinblastine. All three of the drugs block mitosis at the metaphase/anaphase transition, leading to apoptosis. The mechanism of the mitotic block is not known. On the basis of results with purified microtubules and in living interphase cells, we hypothesized that it involves suppression of spindle microtubule dynamics. Here we measured the effects of the three Vinca alkaloids on dynamics of centromeres and spindle kinetochore-microtubules by a novel approach involving quantitative time-lapse confocal microscopy in living mitotic human U2OS cells. Green fluorescent protein-labeled centromere-binding protein B was used to mark centromeres and kinetochore-microtubule plus ends. In controls, pairs of centromeres on sister chromatids alternated under tension between increasing and decreasing separation (stretching and relaxing). All three of the Vinca alkaloids suppressed centromere dynamics similarly at concentrations that block mitosis. At concentrations approximating the IC(50)s for mitotic accumulation (18.8 nM vinflunine, 7.3 nM vinorelbine, and 6.1 nM vinblastine), centromere dynamicity decreased by 44%, 25%, and 26%, respectively, and the time centromeres spent in a paused state increased by 63%, 52%, and 36%, respectively. Centromere relaxation rates, stretching durations, and transition frequencies all decreased. Thus all three of the drugs decreased the normal microtubule-dependent spindle tension at the centromeres/kinetochores, thereby preventing the signal for mitotic checkpoint passage. The strong correlation between suppression of kinetochore microtubule dynamics and mitotic block indicates that the primary mechanism by which the Vinca alkaloids block mitosis is suppression of spindle microtubule dynamics. PMID- 12748303 TI - Role of SODD in regulation of tumor necrosis factor responses. AB - Signaling from tumor necrosis factor receptor type 1 (TNFR1) can elicit potent inflammatory and cytotoxic responses that need to be properly regulated. It was suggested that the silencer of death domains (SODD) protein constitutively associates intracellularly with TNFR1 and inhibits the recruitment of cytoplasmic signaling proteins to TNFR1 to prevent spontaneous aggregation of the cytoplasmic death domains of TNFR1 molecules that are juxtaposed in the absence of ligand stimulation. In this study, we demonstrate that mice lacking SODD produce larger amounts of cytokines in response to in vivo TNF challenge. SODD-deficient macrophages and embryonic fibroblasts also show altered responses to TNF. TNF induced activation of NF-kappaB is accelerated in SODD-deficient cells, but TNF induced c-Jun N-terminal kinase activity is slightly repressed. Interestingly, the apoptotic arm of TNF signaling is not hyperresponsive in the SODD-deficient cells. Together, these results suggest that SODD is critical for the regulation of TNF signaling. PMID- 12748305 TI - Novel vitamin E analogue decreases syngeneic mouse mammary tumor burden and reduces lung metastasis. AB - A nonhydrolyzable ether analogue of RRR-alpha-tocopherol, 2,5,7,8-tetramethyl-2R (4R, 8R, 12-trimethyltridecyl)chroman-6-yloxyacetic acid, called RRR-alpha tocopheryloxyacetic acid or RRR-alpha-tocopherol ether-linked acetic acid analogue (alpha-TEA), exhibits antitumor activity in vitro and in vivo using a syngeneic BALB/c mouse mammary tumor model (line 66 clone 4 stably transfected with green fluorescent protein). Treatment of cells with 5, 10, and 20 micro g/ml alpha-TEA for 3 days produced 6, 34, and 50% apoptosis, respectively, and treatment of cells with 10 micro g/ml for 2, 3, 4, and 5 days produced 20, 35, 47, and 58% apoptosis, respectively. A liposomal formulation of alpha-TEA administered by aerosol reduced s.c. tumor growth and lung metastasis. Alpha-TEA treated animals showed a significant decrease in tumor volumes over 17 days of aerosol treatment (P < 0.001). Forty percent of aerosol as well as untreated control mice had visible, macroscopic lung metastases versus none (0%) of the alpha-TEA-treated mice. On the basis of fluorescence microscopic examination of the surface (top and bottom) of flattened whole left lung lobes, an average of 60 +/- 15 and 102 +/- 17 versus 11 +/- 4 fluorescent microscopic metastases was observed in aerosol control and untreated control versus alpha-TEA-treated animals, respectively. Alpha-TEA formulated in ethanol + peanut oil (5 mg/mouse/day) delivered by gavage did not reduce s.c. primary tumor burden; however, fluorescent microscopic lung metastases were significantly reduced (P < 0.0021). In summary, alpha-TEA formulated in liposomes and delivered by aerosol is a potent antitumor agent and reduces lung metastasis. PMID- 12748306 TI - An anti-tumor necrosis factor-alpha antibody inhibits the development of experimental skin tumors. AB - The proinflammatory cytokine tumor necrosis factor-alpha (TNF-alpha) was originally considered to have activity against malignant disease. However, recent studies suggest TNF-alpha may also act as an endogenous tumor promoter. In the present work, mice deficient in TNF-alpha either genetically (TNF-alpha(-/-)) or after blockade with a neutralizing antibody (cV1q) were used to investigate the role of TNF-alpha in skin tumor development. Papillomas were induced in wild-type (wt) mice after treatment of skin with the initiating agent 9,10-dimethyl-1,2 benzanthracene followed by promotion with 12-O-tetradecanoylphorbol-13-acetate (TPA) for 15 weeks. TNF-alpha(-/-) mice were resistant to papilloma development when compared with wt mice on C57Bl/6J, 129/SvEv, and BALB/c genetic backgrounds. Primary murine keratinocytes (newborn keratinocytes) and skin homogenates were used to characterize TPA-stimulated TNF-alpha expression. TPA induced TNF-alpha protein in newborn keratinocytes in vitro and epidermis in vivo. Neutralization of TNF-alpha protein with cV1q in vivo for 0-15 weeks of promotion significantly decreased skin tumor development after 9,10-dimethyl-1,2-benzanthracene/TPA treatment. cV1q treatment during the early stages of tumor promotion (0-6 weeks) was equally effective. These data suggest that early induction of TNF-alpha is critical for skin tumor promotion. cV1q also reduced TPA-stimulated expression of matrix metalloproteinase 9 and granulocyte macrophage colony-stimulating factor, proteins that are differentially regulated in wt and TNF-alpha(-/-) epidermis. Treatment of the 410.4 transplantable breast carcinoma with cV1q reduced tumor growth in vivo, illustrating that inhibition of tumor growth through neutralization of TNF-alpha is not limited to skin carcinogenesis. These results provide further evidence for procancer actions of TNF-alpha and give some rationale for use of TNF-alpha antagonists in cancer prevention and treatment. PMID- 12748307 TI - Dominant negative signal transducer and activator of transcription 2 (STAT2) protein: stable expression blocks interferon alpha action in skin squamous cell carcinoma cells. AB - We have demonstrated previously that suppression of some or all of the IFN stimulated gene factor 3 (ISGF-3) proteins in skin squamous cell carcinomas is an early event in squamous skin carcinogenesis. This finding led to the hypothesis that suppressed expression of ISGF-3 proteins may lead to reduced IFN responsiveness, which in turn may contribute to skin malignancy by conferring a growth and/or survival advantage. To test this hypothesis, we have developed a skin cell-based model for inhibiting the IFN-alpha signaling pathway through the forced expression of a dominant negative-acting signal transducer and activator of transcription 2 (dnSTAT2) protein. Expression of dnSTAT2 suppressed cell growth inhibition with a pharmacologically achievable concentration (100 IU/ml) of IFN-alpha in the IFN-alpha-sensitive skin squamous cell carcinoma cell line SRB12-p9. dnSTAT2 also suppressed the IFN-alpha-induced phosphorylation of signal transducer and activator of transcription (STAT) 1 and STAT2, which are early events following IFN-alpha treatment, but did not suppress the IFN-gamma-induced phosphorylation of STAT1. Finally, the dnSTAT2 protein suppressed the up regulation of several IFN-alpha-inducible genes that were identified in this system by cDNA microarray screening. We conclude that the cell growth-inhibitory effect of IFN-alpha in skin cells requires an intact STAT2 protein and is therefore mediated by the ISGF-3 complex. These results support STAT2 as an important molecular target for skin cancer chemoprevention. Furthermore, we propose that these dnSTAT2-expressing cells provide a novel in vitro model for the study of type I IFN action in human skin cells. PMID- 12748308 TI - Src family kinase activity is required for signal tranducer and activator of transcription 3 and focal adhesion kinase phosphorylation and vascular endothelial growth factor signaling in vivo and for anchorage-dependent and independent growth of human tumor cells. AB - The Src family kinases (SFKs) Src and Yes are believed to play critical roles in tumor growth, angiogenesis, invasion, and dissemination. Using a panel of highly selective and structurally diverse Src inhibitors, we found that phosphorylation of signal transducer and activator of transcription 3 [STAT3 (Y705)] and focal adhesion kinase [FAK (Y861)] was SFK dependent in cultured human colon, breast, lung, and ovarian tumor cells. These findings were reproduced in vivo in target modulation studies using tumors derived from fibroblasts overexpressing activated Src. Additionally, treatment of mice with multiple Src inhibitors resulted in inhibition of phosphorylation of FAK (Y861) and of a putative Src autophosphorylation epitope (Y419) in HT-29 human colon tumor xenografts. Next we pharmacologically examined the requirement for SFKs in asynchronous proliferation of human tumor cells. At concentrations sufficient to selectively inhibit Src, structurally diverse Src inhibitors inhibited growth of cultured human colon, breast, and lung cells on plastic under low serum conditions. In addition, these compounds inhibited anchorage-independent growth of HT-29 human colon tumor cells in soft agar. The role of SFK activity in vascular endothelial growth factor signaling was also evaluated. Inhibition of SFK signaling using structurally distinct Src inhibitors resulted in complete inhibition of vascular endothelial growth factor-dependent vascular permeability in vivo. These data demonstrate that STAT3 (Y705) and FAK (Y861) phosphoepitopes are SFK-dependent in tumor cells and reveal a requirement for SFK function in tumor cell proliferation and vascular permeability. PMID- 12748309 TI - SU11248 inhibits KIT and platelet-derived growth factor receptor beta in preclinical models of human small cell lung cancer. AB - The purpose of this study was to evaluate the activity of the indolinone kinase inhibitor SU11248 against the receptor tyrosine kinase KIT in vitro and in vivo, examine the role of KIT in small cell lung cancer (SCLC), and anticipate clinical utility of SU11248 in SCLC. SU11248 is an oral, multitargeted tyrosine kinase inhibitor with direct antitumor and antiangiogenic activity through targeting platelet-derived growth factor receptor (PDGFR), vascular endothelial growth factor receptor, KIT, and FLT3 receptors. Treatment of the KIT-expressing SCLC derived NCI-H526 cell line in vitro with SU11248 resulted in dose-dependent inhibition of stem cell factor-stimulated KIT phosphotyrosine levels and proliferation. The biological significance of KIT inhibition was evaluated in vivo by treating mice bearing s.c. NCI-H526 tumors with SU11248 or another structurally unrelated KIT inhibitor, STI571 (Gleevec), which is also known to inhibit Bcr-Abl and PDGFRbeta. SU11248 treatment resulted in significant tumor growth inhibition, whereas inhibition from STI571 treatment was less dramatic. Both compounds reduced phospho-KIT levels in NCI-H526 tumors, with a greater reduction by SU11248, correlating with efficacy. Likewise, phospho-PDGFRbeta levels contributed by tumor stroma and with known involvement in angiogenesis were strongly inhibited by SU11248 and less so by STI571. Because platinum-based chemotherapy is part of the standard of care for SCLC, SU11248 was combined with cisplatin, and significant tumor growth delay was measured compared with either agent alone. These results expand the profile of SU11248 as a KIT signaling inhibitor and suggest that SU11248 may have clinical potential in the treatment of SCLC via direct antitumor activity mediated via KIT as well as tumor angiogenesis via vascular endothelial growth factor receptor FLK1/KDR and PDGFRbeta. PMID- 12748310 TI - Exisulind-induced apoptosis in a non-small cell lung cancer orthotopic lung tumor model augments docetaxel treatment and contributes to increased survival. AB - We reported previously a significant increase in survival of nude rats harboring orthotopic A549 human non-small cell lung cancer tumors after treatment with a combination of exisulind (Sulindac Sulfone) and docetaxel (D. C. Chan, Clin. Cancer Res., 8: 904-912, 2002). The purpose of the current study was to determine the biochemical mechanisms responsible for the increased survival by an analysis of the effects of both drugs on A549 orthotopic lung tumors and A549 cells in culture. Orthotopic A549 rat lung tissue sections from drug-treated rats and A549 cell culture responses to exisulind and docetaxel were compared using multiple apoptosis and proliferation analyses [i.e., terminal deoxynucleotidyl transferase mediated nick end labeling, active caspase 3, the caspase cleavage products cytokeratin 18 and p85 poly(ADP-ribose) polymerase, and Ki-67]. Immunohistochemistry was used to determine cyclic GMP (cGMP) phosphodiesterase (PDE) expression in tumors. The cGMP PDE composition of cultured A549 cells was resolved by DEAE-Trisacryl M chromatography and the pharmacological sensitivity to exisulind, and additional known PDE inhibitors were determined by enzyme activity assays. Exisulind inhibited A549 cell cGMP hydrolysis and induced apoptosis of A549 cells grown in culture. PDE5 and 1 cGMP PDE gene family isoforms identified in cultured cells were highly expressed in orthotopic tumors. The in vivo apoptosis rates within the orthotopic tumors increased 7-8-fold in animals treated with the combination of exisulind and docetaxel. Exisulind increased the in vivo apoptosis rates as a single agent. Docetaxel, but not exisulind, decreased proliferative rates within the tumors. The data indicate that exisulind-induced apoptosis contributed significantly to the increased survival in rats treated with exisulind/docetaxel. The mechanism of exisulind induced apoptosis involves inhibition of cGMP PDEs, and these results are consistent with a cGMP-regulated apoptosis pathway. PMID- 12748311 TI - Near-infrared optical imaging of proteases in cancer. AB - Near-infrared optical imaging is a newer imaging technique that, coupled with sensitive enzymatically specific fluorescent beacons, shows much promise for earlier detection of many cancers and their in situ characterization. On the basis of animal studies demonstrating visualization of micrometastasis-sized tumors and the ability to evaluate therapeutic enzyme inhibition real-time, such imaging may be incorporated in the clinical imaging paradigm in the future, both to improve cancer screening as well as for monitoring therapy in individual patients. This review details some of the related biology, optical probe design, and required hardware, with in vivo cathepsin and matrix metalloprotinease imaging used as examples. PMID- 12748312 TI - Multivoxel magnetic resonance spectroscopy of brain tumors. AB - Magnetic resonance spectroscopy (MRS) is becoming more widely available for clinical applications and is able to provide information about the metabolic properties of regions of normal and abnormal tissue morphology. A critical question for the clinical management of patients with brain tumors is whether multivoxel MRS is able to add new information to the high-quality anatomical data provided by conventional MR imaging techniques and whether this information is relevant for the diagnosis and clinical management of such patients. In this article, the state of the art for acquiring and analyzing multivoxel MRS data is reviewed and placed in context relative to imaging findings for metastatic and primary brain tumors. The MRS data are seen to provide unique information that when combined with high-quality anatomical MR images has implications for defining tumor type and grade, directing biopsy or surgical resection, planning focal radiation or biological therapies, and understanding the mechanisms of success and failure of new treatments. PMID- 12748313 TI - Managing SARS amidst uncertainty. PMID- 12748314 TI - SARS-associated coronavirus. PMID- 12748315 TI - SARS and Carlo Urbani. PMID- 12748316 TI - Images in clinical medicine. Radiologic manifestations of severe acute respiratory syndrome. PMID- 12748317 TI - Heart failure. PMID- 12748319 TI - SARS, the Internet, and the Journal. PMID- 12748318 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 15-2003. A 47-year-old man with waxing and waning pulmonary nodules five years after treatment for testicular seminoma. PMID- 12748320 TI - Platelet storage--time to come in from the cold? PMID- 12748321 TI - The use of corticosteroids in SARS. PMID- 12748323 TI - Pulmonary-artery catheters in high-risk surgical patients. PMID- 12748322 TI - SARS in Northern Vietnam. PMID- 12748324 TI - Peritoneal dialysis and epithelial-to-mesenchymal transition. PMID- 12748325 TI - Outbreaks of infection associated with bronchoscopes. PMID- 12748326 TI - Classification of human papillomavirus. PMID- 12748328 TI - Herpes zoster. PMID- 12748327 TI - Pharmacogenetics. PMID- 12748329 TI - Marked increase in serum IgM during treatment of Waldenstrom's macroglobulinemia with cladribine. PMID- 12748330 TI - C-reactive protein and cardiovascular risk: an update on what is going on in cardiology. PMID- 12748331 TI - Update on interleukin-6 and its role in chronic renal failure. PMID- 12748332 TI - Surgical alternatives to central venous catheters in chronic renal replacement therapy. PMID- 12748333 TI - Skin cancers after transplantation. PMID- 12748334 TI - Reflections on the HEMO study: the American viewpoint. PMID- 12748335 TI - Dose of dialysis, convection and haemodialysis patients outcome--what the HEMO study doesn't tell us: the European viewpoint. PMID- 12748336 TI - Renal replacement therapy in Hungary: the decade of transition. PMID- 12748337 TI - Increased production of chemotactic cytokines and elevated proliferation and expression of intercellular adhesion molecule-1 in rat mesangial cells treated with erythrogenic toxin type B and its precursor isolated from nephritogenic streptococci. AB - BACKGROUND: Previous reports have demonstrated the presence of streptococcal erythrogenic toxin type B (ETB) as well as proliferation and expression of adhesion molecules along with leukocyte infiltrations in biopsies from patients with acute post-streptococcal glomerulonephritis (APSGN). The purpose of the present study was to correlate infiltrative and proliferative events with interactions between ETB or its precursor (ETBP) and intrinsic mesangial cells. METHODS: Rat mesangial cells were cultured with ETB or ETBP (50 micro g/ml) while measuring production of monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-2 (MIP-2) and while examining proliferation and expression of intercellular adhesion molecule-1 (ICAM-1). After 24, 48 and 96 h of incubation, MCP-1 and MIP-2 in culture supernatants were assessed by enzyme linked immunosorbent assay (ELISA). Cells were assessed for proliferation by incorporation of radioactive thymidine and expression of ICAM-1 was measured by indirect immunofluorescence and by cellular ELISA. RESULTS: Compared with controls, treatment with either ETBP or ETB significantly increased MCP-1 and MIP 2 levels in mesangial cell cultures. Mesangial cells also showed elevated proliferation at 96 h of culture when treated with streptococcal proteins. Although production of MCP-1 and MIP-2 was not correlated with proliferation, treatment with ETBP resulted in a significant correlation between MCP-1 production and proliferation. Immunofluorescence studies revealed an increased expression of ICAM-1 in ETBP/ETB-treated mesangial cells. In addition, cellular ELISA studies showed increased absorbance in cultures treated with ETBP/ETB. Finally, low serum concentrations in the culture medium potentiated the stimulatory effect of ETB on MCP-1 production. CONCLUSIONS: Our findings, by demonstrating a role for cationic streptococcal ETB or ETBP in the induction of chemotactic molecules as well as the proliferation and expression of adhesion molecules, delineate an additional possible pathway for the pathogenesis of APSGN. PMID- 12748338 TI - Uraemic medium accelerates proliferation but does not induce apoptosis of endothelial cells in culture. AB - BACKGROUND: Chronic renal failure patients exhibit accelerated atherosclerosis, which is associated with a high incidence of cardiovascular death. We investigated the potential effect of uraemic medium on cell proliferation and apoptosis of endothelial cells in culture (ECs), two key processes in the development of atherosclerosis. Phosphorylation kinetics of the mitogen-activated protein kinase (MAPK) p42/44 and p38 were also evaluated. METHODS: ECs were cultured with growth media supplemented with pooled sera from healthy donors. Semiconfluent ECs were incubated for 24 h with media supplemented with pools of control or uraemic sera. Cell proliferation was assessed through morphometric analysis and by flow cytometry evaluation of cell cycle. To investigate if uraemic medium induces apoptosis in ECs, we used a combination of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) assay and activation of caspase-3 using flow cytometry. Changes in the phosphorylation levels of MAPK were evaluated in cell lysates by western blotting. RESULTS: Exposure to uraemic media caused an alteration in the morphology of ECs, showing irregular shape and size. The number of ECs at S+G(2)M phase in the cell cycle was found to be increased when exposed to uraemic media for 24 h (28.4+/-2.9 vs 20.2+/-2.6% in control ECs). There was a transient increase in levels of phosphorylation of MAPK in both cells, although these levels were significantly higher in ECs exposed to uraemic media, especially after 5 min. In contrast, no signs of apoptosis were observed in ECs incubated with uraemic medium at the conditions applied. CONCLUSIONS: Under our experimental conditions, uraemic medium accelerates proliferation of ECs, but it does not seem to induce apoptosis. The increased proliferation observed could be related to a higher MAPK activity in these cells. Although the enhanced atherosclerosis cannot be explained on the basis of an apoptotic process, the proliferative status could contribute to intimal proliferation, which is considered to be an earlier step in the development of atherosclerosis. PMID- 12748340 TI - Diversity in rat tissue accumulation of vitamin B12 supports a distinct role for the kidney in vitamin B12 homeostasis. AB - BACKGROUND: Vitamin B(12) in plasma is complexed to the carrier proteins transcobalamin (TC) and haptocorrin. The TC-B(12) complex is filtered in the glomeruli and reabsorbed in the renal tubules by receptor-mediated endocytosis, providing a route for a significant renal accumulation of vitamin B(12). The present study investigates the role of the rodent kidney in B(12) homeostasis by examining the distribution of vitamin B(12) in rats during vitamin B(12) depletion or B(12) load, and compares kidney accumulation with the vitamin distribution in other tissues including brain, liver, testes, intestine, spleen and plasma. METHODS: Fifteen rats were fed on a diet containing different concentrations of B(12) supplemented with s.c. injections of B(12). Twenty four hours prior to sacrifice, all animals were injected with [(57)Co]B(12). The vitamin contents of kidneys, liver, spleen, brain, testis, intestine, skeletal muscle, serum and urine were analysed. Both total tissue vitamin B(12) accumulation and [(57)Co]B(12) were determined to compare steady-state B(12) and the distribution of an acutely injected dose. In the kidney, free and protein bound B(12) was determined by gel filtration. RESULTS: The rat kidneys accumulated more B(12) during normal and loaded conditions than any other tissue. A 110-fold increase in vitamin content was observed from the deficient to the loaded conditions in the kidney compared with a 3.5-fold increase in the liver. In contrast to all other organs, significantly smaller amounts of acutely injected B(12) accumulated in the kidneys in the vitamin-deprived state compared with both the normal and the vitamin-loaded condition. CONCLUSIONS: The present study suggests a significant role for the rodent kidney in vitamin B(12) metabolism. We propose a model for rat tissue uptake consistent with the presence of two different TC-B(12) receptors and renal uptake following filtration of TC B(12) in the glomeruli. The presented model allows for the reduced renal uptake and accumulation in vitamin-deprived conditions, thus reserving the vitamin for other tissues, including nerve tissue and bone marrow, which are more sensitive to vitamin B(12) deficiency. PMID- 12748339 TI - Different effects of amino acid-based and glucose-based dialysate from peritoneal dialysis patients on mesothelial cell ultrastructure and function. AB - BACKGROUND: Peritoneal dialysis fluid (PDF) containing amino acids has been introduced recently aiming to improve the nutritional status of PD patients. Dextrose-based PDFs have been implicated in progressive functional and structural deterioration of the peritoneal membrane. Limited data are currently available regarding the effect of amino acid-based PDF on the function and ultrastructure of human peritoneal mesothelial cells (HPMCs), which play a critical role in peritoneal membrane pathophysiology. METHODS: We investigated the effects of two commercially available PDFs, which utilized dextrose (1.5% Dianeal) or amino acids (1.1% Nutrineal) as the osmotic agent, obtained from patients after a 4 h dwell, on HPMC proliferation (MTT assay and cell counting) and viability [lactate dehydrogenase (LDH)release], interleukin-6 (IL-6) secretion (commercial enzyme linked immunosorbent assay) and ultrastructure (scanning and transmission electron microscopy). RESULTS: Exposure of HPMCs to 1.5% Dianeal reduced cell proliferation, total cellular protein synthesis, IL-6 secretion and cell attachment, but prolonged the cell doubling time on recovery, and increased LDH release (P<0.001, P<0.001, P<0.0001, P<0.0001, P<0.001 and P<0.001, respectively). The 1.1% Nutrineal reduced HPMC proliferation (P<0.001) and increased IL-6 secretion (P<0.0001), but did not affect cell attachment, LDH release, protein synthesis or cell doubling time. Ultrastructural studies of HPMCs exposed to Dianeal showed cell flattening, increased cell surface area, reduced microvilli, and intracellular organelles compatible with dysfunctional mitochondria. In contrast, the ultrastructural morphology of HPMCs was relatively preserved after incubation with Nutrineal. CONCLUSIONS: Our results showed that HPMC ultrastructure, viability and protein synthesis were better preserved with amino acid-based PDF, compared with conventional dextrose-based PDF. The significance of IL-6 induction by Nutrineal remains to be elucidated. PMID- 12748341 TI - Dynamics of secretion and metabolism of PTH during hypo- and hypercalcaemia in the dog as determined by the 'intact' and 'whole' PTH assays. AB - BACKGROUND: Recent evidence has shown that the assay for 'intact' parathyroid hormone (I-PTH) not only reacts with 1-84 PTH but also with large non-1-84 PTH fragments, most of which is probably 7-84 PTH. As a result, an assay specific for 1-84 PTH named 'whole' PTH (W-PTH) has been developed. The present study was designed: (i) to determine whether the W-PTH assay reliably measures PTH values in the dog; (ii) to evaluate differences between the W-PTH and I-PTH assays during hypo- and hypercalcaemia; and (iii) to assess the peripheral metabolism of W-PTH and I-PTH. METHODS: In normal dogs, hypocalcaemia was induced by EDTA infusion and was followed with a 90 min hypocalcaemic clamp. Hypercalcaemia was induced with a calcium infusion. RESULTS: I-PTH and W-PTH values increased from 36+/-8 and 13+/-3 pg/ml (P=0.01) at baseline to a maximum of 158+/-40 and 62+/-15 pg/ml (P=0.02 vs I-PTH) during hypocalcaemia. The W-PTH/I-PTH ratio, 38+/-4% at baseline, did not change during the induction of hypocalcaemia, but sustained hypocalcaemia increased (P<0.05) this ratio. During hypercalcaemia, maximal suppression for I-PTH was 2.0+/-0.5 and only 5.7+/-0.6 pg/ml for W-PTH, due to a decreased sensitivity of the W-PTH assay at values <5 pg/ml. The disappearance rate of PTH was determined in five additional dogs which underwent a parathyroidectomy (PTX). At 2.5 min after PTX, W-PTH was metabolized more rapidly, with a value of 25+/-2% of the pre-PTX value vs 30+/-3% for I-PTH (P<0.05). CONCLUSIONS: (i) The W-PTH/I-PTH ratio is less in the normal dog than in the normal human, suggesting that the percentage of non-1-84 PTH measured with the I-PTH assay is greater in normal dogs than in normal humans; (ii) the lack of change in the W-PTH/I-PTH ratio during acute hypocalcaemia is different from the situation observed in humans; and (iii) the dog appears to be a good model to study I-PTH and W-PTH assays during hypocalcaemia. PMID- 12748342 TI - Tonsillar IgA1 as a possible source of hypoglycosylated IgA1 in the serum of IgA nephropathy patients. AB - BACKGROUND: There are many reports of incompletely glycosylated O-linked oligosaccharides on the IgA1 hinge region in certain IgA nephropathy patients. In addition, other reports have noted a relationship between tonsillectomy and IgA nephropathy. METHODS: Immunoglobulins from extracts of tonsillectomized tissue and other sources were analysed by isoelectric focusing (IEF) and by enzyme linked immunosorbent assay (ELISA). RESULTS: The IEF profile of tonsillar IgA differed from that of serum IgA and it was enriched in cationic IgA. However, extracts from tonsillitis controls and IgA nephropathy patients exhibited profiles that were very similar. Enzymatic removal of sialic acid induced a shift of the peaks to the cathode side. The profiles of IgA from treated tonsillar extract and treated serum were closely overlapped. In addition, asialo Galbeta1,3GalNAc was clearly present in cationic IgA from tonsillar extract and in aberrant IgA1 from serum following enzymatic transfer of sialic acid to IgA1. Serum IgA also contained partly sialylated IgA1. Quantitative analysis of IgA and IgG in the extracts indicated that IgA was significantly higher, whereas IgG was significantly lower in IgA nephropathy patients. CONCLUSIONS: We found that the IgA1 produced in tonsillar tissue differed from serum IgA1. Furthermore, an overproduction of asialo IgA1 resulted from the disordered balance between IgA- and IgG-producing cells in the tonsils from the IgA nephropathy patient. Although it is unclear how such asialo IgA1 molecules are transferred from tonsil tissue to serum, a tonsillar source may produce a few micrograms of aberrant IgA1 that then appears in serum. PMID- 12748343 TI - Effect of losartan and amlodipine on proteinuria and transforming growth factor beta1 in patients with IgA nephropathy. AB - BACKGROUND: Transforming growth factor-beta1 (TGF-beta1) is the major profibrotic cytokine involved in many renal diseases, and urinary TGF-beta1 reflects intrarenal TGF-beta1 production. Urinary TGF-beta1 excretion is reported to be significantly increased in patients with immunoglobulin A (IgA) nephropathy. The aim of the present study was to compare the effects of losartan and amlodipine on proteinuria, as well as on serum and urine TGF-beta1 levels in IgA nephropathy patients with hypertension and proteinuria. METHODS: The initial 4 week washout period was followed by 12 weeks of active treatment, in which patients were randomized to once-daily treatment with losartan 50 mg (group 1, n=20) or amlodipine 5 mg (group 2, n=16). Urinary protein and TGF-beta1 excretion, serum TGF-beta1 and other clinical parameters were determined at baseline and during 12 weeks of active treatment. RESULTS: Both treatments controlled blood pressure (BP) to a similar degree, and renal function and other biochemical parameters did not change during the study period. Urinary protein and TGF-beta1 excretions were significantly elevated in IgA nephropathy patients. Losartan significantly reduced urinary protein (from 2.3+/-1.5 g/day at baseline to 1.2+/-1.5 g/day at 12 weeks, P<0.05) and urinary TGF-beta1 excretion (from 31.2+/-14.0 pg/mg creatinine at baseline to 22.1+/-13.5 pg/mg creatinine at 12 weeks, P<0.05). In contrast, amlodipine had no affect on urinary protein and TGF-beta1 excretion. Both losartan and amlodipine failed to reduce serum TGF-beta1 levels. CONCLUSION: Losartan and amlodipine, with similar control of BP, showed different effects on urine protein or TGF-beta1 excretion. Whereas losartan improved both urinary parameters, amlodipine did not. These differences might be important for the management of IgA nephropathy. PMID- 12748344 TI - Novel COL4A4 splice defect and in-frame deletion in a large consanguine family as a genetic link between benign familial haematuria and autosomal Alport syndrome. AB - BACKGROUND: Alport syndrome (AS) is a common hereditary cause for end-stage renal failure due to a defect in type IV collagen genes. The molecular pathogenesis of benign familial haematuria (BFH) is not fully understood. Evidence from linkage analyses and mutation studies point to a role of the COL4A3/COL4A4 genes. The present study describes molecular changes of the COL4A4 gene that cause both diseases: autosomal recessive AS and BFH in a consanguine family with a 400-year old history of haematuria. METHODS: RNA and DNA were isolated and analysed by RT PCR, PCR, DNA and cDNA sequencing, and Southern blotting. Evaluation of family members comprised creatinine clearence, urine analysis, audiometry and past medical history. RESULTS: Forefathers of this family moved to a German village in the 17th century. Sporadic episodes of macrohaematuria have been reported ever since. Numerous family members with haematuria including the parents of the index family were heterozygous for a splice defect eliminating exon 25 from the alpha4(IV) cDNA. The daughter (15 years old, creatinine clearence 27 ml/min, proteinuria 5 g/day, hearing loss) was homozygous for the mutation, while the son (22 years old, creatinine clearance 68 ml/min, proteinuria 11 g/day, hearing loss, splitted and thickened glomerular basement membrane) was heterozygous. Further analysis showed a second mutation, an 18 bp in-frame deletion in exon 25, for which numerous family members were heterozygous, and both children were homozygous. CONCLUSIONS: The COL4A4 splice defect causes BFH-phenotype in heterozygous, and AS in homozygous state. The clinical spectrum of heterozygous individuals reaches from macrohaematuria, intermittent microhaematuria to isolated deafness. The 18 bp in-frame deletion aggravates the phenotype in the compound heterozygous son. These results give further evidence that BFH and autosomal AS are in fact both type IV collagen diseases. PMID- 12748345 TI - A retrospective 5-year study in Moldova of acute renal failure due to leptospirosis: 58 cases and a review of the literature. AB - BACKGROUND: Renal involvement [as acute renal failure (ARF)] is a prominent feature of both mild and severe leptospirosis-a re-emerging infectious disease. Few large series describe in detail clinical and laboratory features of cases with ARF and their outcome. METHODS: We performed a retrospective analysis (1997 2001) of all consecutive, serological confirmed leptospirosis cases with ARF (n=58, 53 male, age 44+/-13 years, rural residents=31%, animal contact=88%. RESULTS: Clinical manifestations (>50% prevalence): oliguria 95%, fever and jaundice 93%, nausea and vomiting 83%, haemorrhagic diathesis 80%, headache, hepatomegaly 76%, myalgias, abdominal pain 70%, hypotension 62%, disturbed consciousness 50%. A pattern of multiple organ failure (MOF) was frequent: ARF together with hepatic failure in 72%, respiratory failure in 38%, circulatory failure in 33%, pancreatitis in 25% and rhabdomyolysis in 5% of cases. Renal dysfunction: 35% of cases had a renal K(+)-wasting defect and 43% a FE(Na)(+)>1% and low-osmolarity urine despite volume depletion. Haematuria was encountered in 12 and mild proteinuria in 10 subjects. OUTCOME: 26% deaths, 64% normal hepatic and renal function at 90 days from presentation (however 29% maintained the initial tubular defect), 10% persistent mild renal failure. All deceased patients had, beside ARF, at least two other organ failures, affected consciousness, and haemorrhagic diathesis vs a prevalence for the above features of only 34, 33, and 72%, respectively, in the survivors group (P<0.05). CONCLUSIONS: Leptospirosis presenting with ARF is a severe disease, frequently leading to MOF and to death in one-third of the patients. In particular, the haemorrhagic diathesis and cerebral involvement are markers for unfavourable patient and renal outcomes. PMID- 12748346 TI - High serum concentrations of the acyclovir main metabolite 9 carboxymethoxymethylguanine in renal failure patients with acyclovir-related neuropsychiatric side effects: an observational study. AB - BACKGROUND: Acyclovir (ACV) has been used for over two decades to treat herpes virus infections. Serious neurological adverse side effects have occurred during ACV treatment in patients with renal failure, but the cause of the symptoms remains unknown. We hypothesized that increased concentrations of the ACV main metabolite 9-carboxymethoxymethylguanine (CMMG) correlated to these symptoms. METHODS: We conducted an observational study from 1991 to mid 1999 based on samples sent for analysis of ACV concentration from various hospital departments in Sweden. Patients with neuropsychiatric symptoms (NS+, n=49) were compared with patients without symptoms (NS-, n=44). ACV and CMMG concentrations were analysed by HPLC. Medical records were analysed for symptoms and compared with pertinent cases identified from Medline. RESULTS: The serum CMMG levels were significantly higher in the NS+ group (mean=34.1 micro mol/l, 95% confidence interval 23.4 46.1) compared with the NS- group (mean=4.7 micro mol/l, 95% confidence interval 3.3-6.6; P<0.001). CMMG was the strongest predictor in a receiver-operating characteristics curve analysis (ROC), based on 77 patients, of ACV-related neuropsychiatric symptoms. The ROC curve for CMMG demonstrated that neuropsychiatric symptoms could be predicted with a sensitivity of 91% and a specificity of 93% with the use of a cut-off value of 10.8 micro mol/l of CMMG. Thirty-five of 49 patients in the NS+ group showed levels exceeding this concentration compared with only three of 44 of patients in the NS- group (P<0.001). ACV exposure, ACV concentration, creatinine clearance and creatinine concentration were weaker but statistically significant predictors. Haemodialysis reduced CMMG and ACV levels and relieved the symptoms. CONCLUSIONS: The determination of CMMG levels in serum may be a useful tool in supporting the diagnosis of ACV-associated neuropsychiatric symptoms. Furthermore, the monitoring of CMMG levels may prevent the emergence of symptoms. PMID- 12748347 TI - PAI-1 4G/5G and ACE I/D gene polymorphisms and the occurrence of myocardial infarction in patients on intermittent dialysis. AB - BACKGROUND: Myocardial infarction (MI) is a leading cause of death, particularly in high-risk settings such as uraemia, in which it is not yet known to what extent genetic factors contribute to the overall risk of MI. We have prospectively evaluated the effect of plasminogen activator inhibitor-1 (PAI-1) 4G/5G and angiotensin-converting enzyme (ACE) insertion/deletion (I/D) polymorphisms on the occurrence of MI in uraemics. METHODS: All patients undergoing intermittent dialysis in an Italian district were enrolled as subjects. From the same area, 1307 individuals served as controls. Genomic DNA was obtained and ACE I/D and PAI-1 4G/5G gene polymorphisms were determined. After a baseline evaluation, patients were followed for 28.8+/-9.8 months. MIs and other causes of death were recorded. RESULTS: A total of 461 patients (417 on haemodialysis and 44 on peritoneal dialysis) were investigated. At entry, their mean age was 58.2+/-16.2 years and dialytic age was 82+/-69 months. Genotype frequencies were not different between controls and uraemics and, in the latter group, between patients with or without cardiovascular diseases at baseline evaluation. During the follow-up, 22 fatal and 16 non-fatal MIs were recorded (mean incidence 1.99 and 1.45%/year, respectively). The adjusted risk of fatal and total MI was related to the presence at entry of a history of MI [hazard ratios (HR) 4.3; 95% confidence interval (CI) 1.5-12.0 and HR 6.8; 95% CI: 3.3 14.0, respectively] and to the PAI-1 4/4 genotype (HR 2.8; 95% CI 1.2-6.9 and HR 2.1; 95% CI 1.1-4.2, respectively). CONCLUSIONS: In end-stage renal disease, PAI 1 4G/5G gene polymorphism may have a significant role in the occurrence of fatal and non-fatal MI. PMID- 12748348 TI - Prevalence of atherosclerotic renal artery stenosis in patients starting dialysis. AB - BACKGROUND: Atherosclerotic renal artery stenosis (ARAS) can lead to end-stage renal failure (ESRF). We determined the prevalence of ARAS in patients 45 years of age or older starting renal replacement therapy. METHODS: Forty-nine of 80 consecutive patients (37 males, 12 females) starting renal replacement therapy in our centre gave informed consent and underwent spiral computed tomographic angiography of their renal arteries. A renal artery diameter reduction of 50% or more assessed by two radiologists was considered as a significant stenosis. RESULTS: Twenty of 49 patients (41%) had an ARAS, and in eight cases (16%) this was bilateral or unilateral with a single kidney. Women were more likely to have an ARAS than men; 75 (9/12) vs 30% (11/37, P<0.01). However, relatively more women declined participation. Non-participants and participants did not differ in respect to other relevant clinical data. Nonetheless, findings in these patients would be negative, the prevalence of ARAS would still be 31% in women and 22% in men (NS). In 13 patients with ARAS the registered diagnosis of ESRF either was hypertension, renovascular disease or unknown. Assuming that in these patients atherosclerotic renovascular disease was the cause of renal failure, a total of 13 patients (13/49, 27%) entered the dialysis programme because of this problem. CONCLUSIONS: These results suggest that ARAS is an important cause of ESRF. PMID- 12748349 TI - Assessment of vascular calcification in ESRD patients using spiral CT. AB - BACKGROUND: Dialysis patients have increased vascular calcification of the coronary arteries and aorta by electron beam CT scan. The purpose of the present study was to utilize an alternative machine, spiral CT, to assess calcification in end-stage renal disease (ESRD) patients. METHODS: Two groups of patients with ESRD were evaluated: group 1, those receiving a renal transplant (n=38); and group 2, those remaining on dialysis (n=33). All patients underwent quad-slice spiral CT with retrospective gating to evaluate coronary artery and aorta calcification scores. Both area (Agatston method) and volume calculations were utilized, with retrospective gating in all but 16 subjects. Laboratory tests, medications and clinical characteristics were analysed. RESULTS: Using spiral CT, the intra-reader variability for coronary artery calcification (after correction for very low scores) was 0.9% mean / 0% median using the area (Agatston method) and 2.9% mean / 0% median using volume calculations. Group 1 patients were younger, more likely to be Caucasian and on peritoneal dialysis, had lower serum calcium and higher C-reactive protein levels than group 2. In patients without vs those with coronary artery calcification, only longer duration of dialysis (34+/ 64 vs 55+/-50 months, P=0.004; r=0.39, P=0.005) and increasing age (39+/-13 vs 54+/-10 years, P<0.001; r=0.29, P=0.039) were associated, whereas only increasing age was associated with aorta calcification. CONCLUSION: In ESRD patients, the factors correlating with coronary calcification were duration of dialysis and advancing age, whereas only age correlated with aorta calcification. Spiral CT offers an alternative technique for the assessment of these changes. PMID- 12748350 TI - Spectrum of renal bone disease in end-stage renal failure patients not yet on dialysis. AB - BACKGROUND: During the last few years the spectrum of renal osteodystrophy (ROD) in dialysis patients has been studied thoroughly and the prevalence of the various types of ROD has changed considerably. Whereas until a decade ago most patients presented with secondary hyperparathyroidism (HPTH), adynamic bone (ABD) has become the most common lesion within the dialysis population over the last few years. Much less is known about the spectrum of ROD in end-stage renal failure (ESRF) patients not yet on dialysis. METHODS: Transiliac bone biopsies were taken in an unselected group of 84 ESRF patients (44 male, age 54+/-12 years) before enrolment in a dialysis programme. All patients were recruited within a time period of 10 months from various centres (n=18) in Macedonia. Calcium carbonate was the only prescribed medication in patients followed up by the outpatient clinic. RESULTS: HPTH was found in only 9% of the patients, whilst ABD appeared to be the most frequent renal bone disease as it was observed in 23% of the cases next to normal bone (38%). A relatively high number of patients (n=10; 12%) fulfilled the criteria of osteomalacia (OM). Mixed osteodystrophy (MX) was diagnosed in 18% of the subjects. There was no significant difference between groups in age, creatinine, or serum and bone strontium and aluminium levels. Patient characteristics associated with ABD included male gender and diabetes, whilst OM was associated with older age (>58 years). CONCLUSIONS: In an unselected population of ESRF patients already, 62% of them have an abnormal bone histology. ABD is the most prevalent type of ROD in this population. In the absence of aluminium or strontium accumulation the relatively high prevalence of a low bone turnover as expressed by either normal bone or ABD and OM is striking. PMID- 12748351 TI - White blood cells as a novel mortality predictor in haemodialysis patients. AB - BACKGROUND: Many conventional cardiovascular risk factors in the general population are not as predictive in end-stage renal disease (ESRD). As absolute neutrophil count and total white blood cell (WBC) count are associated with adverse cardiovascular outcomes and all-cause mortality, this analysis was undertaken to explore the associations of WBC variables with mortality risk in ESRD. METHODS: Of a total study population of 44 114 ESRD patients receiving haemodialysis during 1998 at facilities operated by Fresenius Medical Care, North America, 25 661 patients who underwent differential white cell count and had complete follow-up were included. Information on case mix (age, gender, race), clinical (diabetes, body mass index), and laboratory variables (haematocrit, albumin, creatinine, potassium, calcium, phosphorus, bicarbonate, ferritin, transferrin saturation and differential WBC count) was obtained. Associations between lymphocyte count, neutrophil count and demographic and clinical variables were examined using linear regression. Associations between WBC variables and survival were estimated using Cox proportional hazard regression. RESULTS: A higher lymphocyte count was associated with higher serum albumin and creatinine, lower age and black race. High neutrophil count was associated with lower serum albumin and creatinine, younger age and white race (all Ps <0.0001). Cox proportional hazard regression showed an increased lymphocyte count was associated with reduced mortality risk [HR 0.86 (0.83-0.89) per 500/ml increase in lymphocyte count] and an increased neutrophil count was associated with increased mortality risk [HR 1.08 (1.06-1.09) per 1000/ml increase in neutrophil count]. CONCLUSIONS: An increased neutrophil count is strongly associated with, and reduced lymphocyte count associated less strongly with, many surrogates of both malnutrition and inflammation. An increased neutrophil count and reduced lymphocyte count are independent predictors of increased mortality risk in haemodialysis patients. PMID- 12748352 TI - Reducing vascular access morbidity: a comparative trial of two vascular access monitoring strategies. AB - BACKGROUND: Thrombosis is the primary cause of access failure in polytetrafluoroethylene grafts and arteriovenous fistulas. It can lead to significant patient and access morbidity and mortality, and is difficult to prevent medically. Intervention is largely limited to maximizing access patency by detecting culprit lesions early and intervening with angioplasty or surgical revision. The most efficacious monitoring strategy is undetermined. METHODS: This 3 year prospective study took advantage of a change in monitoring strategy used in a large dialysis centre to compare the efficacy of two methods used to monitor grafts and fistulas in order to prevent access thrombosis. Accesses were monitored using Duplex ultrasonography in year 1, while the saline ultrasound dilution technique (Transonic) became the primary monitoring strategy in year 3 (year 2 was a transition year). Risk factors for thrombosis were determined using multivariate survival analysis, and the performance of Duplex ultrasonography and Transonic monitoring was assessed. RESULTS: A total of 303 656 access days at risk were assessed, with 344, 385 and 425 accesses in years 1, 2 and 3, respectively. The total thrombosis rate was 1.01/1000 access days in year 1 compared with 0.66/1000 access days in year 3. This was accomplished despite a reduction in procedure rates of 55% for angiograms, 13% for angioplasties and 31% for thrombolysis. CONCLUSION: Low flow rates detected using Transonic monitoring were associated with increased thrombosis, while stenosis detected using Duplex ultrasonography was not a strong predictor of incipient thrombosis; however, these different access characteristics were compared using monitoring techniques that may be ideal in different clinical situations. PMID- 12748353 TI - Patterns of CD4/CD8 T-cell ratio in dialysis effluents predict the long-term outcome of peritonitis in patients undergoing peritoneal dialysis. AB - BACKGROUND: The peritoneal immune compartment is a microenvironment with a particular T-cell repertoire and susceptible to local inflammation. To clarify the role of T lymphocytes in peritoneal immunity, the changes in T-cell subpopulations in peritoneal dialysis effluents (PDEs), and their influence on the response to the treatment of peritonitis and on its prognosis were studied in patients undergoing long-term, continuous ambulatory peritoneal dialysis (CAPD). METHODS: A cohort of 36 patients treated with CAPD and who had histories of peritonitis were divided into a group with rapid and a group with delayed response to antibiotics, and were followed for 3 years. CD4/CD8 T-cell ratios, T cell cytokine mRNA expression patterns and transforming growth factor-beta1 (TGF beta1) concentrations were examined in PDE during bouts of peritonitis. The change in 4 h D/P creatinine during the peritoneal equilibration test (PET) between year 0 and year 3 was expressed as deltaD/P creatinine. RESULTS: The serial changes in T-cell subsets in PDE during peritonitis showed two patterns: (i) pattern 1, manifest as a progressive increase in the CD4/CD8 ratio, and associated with a rapid response to treatment; and (ii) pattern 2, manifest as a progressive decrease in the CD4/CD8 ratio, and associated with a delayed response to treatment. The major T-cell phenotypes in PDE during peritonitis were Th1 CD4(+) and Tc2-CD8(+), determined by cloning techniques, RT-PCR and double immunofluorescence staining. TGF-beta1 in the effluent was undetectable in pattern 1 after 7-8 days, but remained detectable at 2 weeks in pattern 2. Pattern 2 patients had a significantly greater decrease (deltaD/P creatinine: 0.198+/-0.086) in solute transport than pattern 1 patients (deltaD/P creatinine: 0.036+/-0.077, P<0.05). CONCLUSIONS: These results suggest that a progressive decrease of the CD4/CD8 ratio in PDE correlates with a persistent expression of TGF-beta1, and plays a pathogenetic role in the evolution of peritonitis, PET deterioration and peritoneal fibrosis. Therefore, patterns of CD4/CD8 T-cell ratio in PDE may predict clinical outcomes of peritonitis in CAPD patients. PMID- 12748354 TI - Incidence of polyomavirus-nephropathy in renal allografts: influence of modern immunosuppressive drugs. AB - BACKGROUND: In recent years an increasing number of cases with polyomavirus (PV) nephropathy after renal transplantation were reported from several transplant centres. New, highly potent immunosuppressive drugs like tacrolimus or mycophenolate mofetil were accused as risk factors for this increase. However, data about the incidence of PV-nephropathy in correlation to different immunosuppressive therapy concepts are lacking. METHODS: All renal transplant biopsies performed at Hannover Medical School between 1999 and 2001 (n=1276) were immunohistochemically screened for the presence of PV-specific proteins. The results were correlated to the different immunosuppressive therapy protocols and patients with PV-nephropathy were compared with a matched control group. RESULTS: PV-nephropathy was found in <1% of all investigated allograft biopsies (11/1276) and in approximately 1% of all patients (7/638), respectively. All patients being immunohistochemically positive for PV-specific proteins also showed the typical morphological changes of PV-nephropathy. Four out of seven patients with PV nephropathy were under triple immunosuppression comprising tacrolimus and mycophenolate mofetil. Under this immunosuppressive therapy protocol an eight times higher incidence and a 13 times higher risk (multivariate odds ratio 12.7) of PV-nephropathy was observed in our patients compared with the control group. CONCLUSIONS: PV-nephropathy is a rare but serious complication after renal transplantation. A small group of patients under intensive immunosuppression comprising tacrolimus in combination with mycophenolate mofetil has a significantly increased risk of acquiring this deleterious complication. PMID- 12748355 TI - Mycophenolate mofetil is associated with less death with function than azathioprine in cadaveric renal transplantation. AB - BACKGROUND: A previous study has argued that mycophenolate mofetil (MMF) is associated with a reduced incidence of death with function when compared to azathioprine (AZA) in cadaveric renal transplantation. This study was designed to verify this result because methodological issues bring these findings into question. METHODS: The data used in this study was derived from records of renal transplants performed in 1995 and 1996 as recorded in the UNOS Scientific Renal Transplant Registry and supplied by the United States Renal Data System (USRDS). Univariate and multivariate survival analysis was used to compare rates of death with function. Covariate characteristics of the donor, recipient, procedures, early outcomes and the transplant centre were considered. RESULTS: 12,251 recipients of cadaveric renal transplants were identified as having received either MMF or AZA, but not both. The relative risk of death with function calculated by the Kaplan-Meier method was 21% less for MMF patients (P=0.005). MMF had from 21% (P=0.008) to 24% (P=0.001) reductions in relative risk by multivariate methods. CONCLUSIONS: The use of MMF is associated with a reduction in the incidence of death with a functioning graft in cadaveric renal transplantation. These results verify previous analyses. PMID- 12748356 TI - A compartmental pharmacokinetic model of cyclosporin and its predictive performance after Bayesian estimation in kidney and simultaneous pancreas-kidney transplant recipients. AB - BACKGROUND: Therapeutic drug monitoring of cyclosporin A (CsA) is an obvious necessity because of its unpredictable absorption and narrow therapeutic window. The use of limited sampling models (LSMs) has improved the estimation of the systemic exposure [area under curve (AUC)] compared with C(0h) monitoring, but these equations are rigid and not reliable in patients with an abnormal absorption profile. We developed and validated a limited sampling (t=0, 2 and 3 h) strategy, based on a compartmental population pharmacokinetic (PK) model for CsA after kidney transplantation alone (KTA) and simultaneous pancreas-kidney transplant (SPKT) recipients, a group of patients with unpredictable absorption kinetics. METHODS: A two-compartment model with lag time and first-order absorption was calculated using a PK software package from data of 20 KTA and SPKT recipients and validated prospectively in 20 KTA and 20 SPKT recipients. Calculated population PK parameters were individualized for each of the remaining 40 patients based on their CsA dosing and on one or a combination of measured CsA blood concentrations using the Bayesian fitting method. AUCs were calculated from individualized PK parameters. AUCs were also calculated using previously published LSMs. Relationships between AUCs calculated by the models and the 'golden standard' AUC (trapezoidal rule) were investigated by Pearson correlation test. RESULTS AND CONCLUSIONS: A population two-compartment model is presented to reliably estimate the CsA AUC in KTA and SPKT recipients. The performance of the model to estimate the AUC is comparable to the performance of two published LSMs in KTA patients, but markedly better in SPKT patients. Combined with Bayesian fitting, the model is very flexible since sampling times are not rigid and can be varied as long as dosing and sampling times are recorded accurately. The model has already proven to be clinically useful and is currently used to further investigate CsA in an integrated pharmacokinetic/pharmacodynamic model. PMID- 12748358 TI - Symptomatic lactic acidosis due to relapse of T-cell acute lymphoblastic leukaemia in the kidney. PMID- 12748357 TI - Predictors of serum creatinine in haemodialysis patients: a cross-sectional analysis. AB - BACKGROUND: C-reactive protein (CRP) levels, an acute phase response index, predict cardiovascular outcome and are inversely related to visceral proteins, including albuminaemia in haemodialysis patients. Less definite is the relationship between inflammation and markers of somatic proteins such as serum creatinine in such patients. To explore these questions, a cross-sectional analysis of potential predictors of serum creatinine was performed. METHODS: One hundred and seventy-nine prevalent haemodialysis patients as of June 2001 were included in the cohort. Midweek pre-dialysis blood samples were collected during the months of June, September through to December 2001 inclusive, and determinations of serum urea (urease method), creatinine (alkaline picrate method) and CRP levels by means of a high sensitivity immunonephelometric method were performed. Furthermore, pre- and post-dialysis body weights were recorded and 2 min post-dialysis serum urea levels were determined three times. They were utilized for the calculation of single pool Kt/V and of normalized protein catabolic rate (nPCR). Each of the data represents the mean of three determinations made every 3 months in the study period. RESULTS: The analysis of multivariate linear regression was able to validate our model characterized by a dependent variable, serum creatinine and four independent variables (age, CRP, Kt/V and nPCR) (R(2)=0.60; F=24.10; P<0.00001; SE=1.94). Age (-0.08 mg/dl decrease in serum creatinine per 1-year increase in age), Kt/V (-0.25 mg/dl decrease in serum creatinine per 0.1 increase in Kt/V) and nPCR (0.10 mg/dl increase in serum creatinine per 0.1 g protein/kg/day increase in nPCR) were independently predictive of serum creatinine (P<0.00001). CRP and dialysis vintage did not predict serum creatinine. Stratifying the patients for the effects of CRP, only CRP values 4 mg/l were not. A further insight was given by the stratification of the patients for the effects of the interquartile ranges of CRP: it showed a progressive and statistically significant reduction of beta-coefficient inversely related to the increasing CRP values (P=0.003). Thus, the nature of the correlation between CRP and serum creatinine changes with increasing CRP values: from being a direct one, it shows a trend towards a transformation into an indirect one with beta=0 at a CRP value of approximately 9 mg/l. However, this indirect relationship does not reach statistical significance. CONCLUSIONS: The present cross-sectional study suggests that the activation of acute phase response does not influence creatinine metabolism in haemodialysis patients; in contrast, age, Kt/V and nPCR predict serum creatinine levels. Larger prospective trials are needed to achieve a definitive answer about the relationship between somatic proteins, acute phase response activation and nutrition in dialysis patients. PMID- 12748359 TI - Solving electrolyte disturbances with the Ehrlich reagent. PMID- 12748360 TI - A young patient with unexplained acute hepatorenal dysfunction. PMID- 12748361 TI - Painful haematuria following central venous catheter infection. PMID- 12748362 TI - Fistulizing TB peritonitis during CAPD. PMID- 12748363 TI - Initiation of dialysis: is the problem solved by NECOSAD? PMID- 12748365 TI - Pitfalls of the glucose pump test for access flow measurements. PMID- 12748367 TI - Aldactone therapy in a peritoneal dialysis patient. PMID- 12748369 TI - Aldactone therapy in a peritoneal dialysis patient. PMID- 12748370 TI - Mycophenolate mofetil in treatment of idiopathic stages III-IV membranous nephropathy. PMID- 12748371 TI - About the pleiotropic effects of statins in human. PMID- 12748372 TI - Acute renal failure in severe leptospirosis. PMID- 12748374 TI - The living kidney donor: giving life, avoiding harm. PMID- 12748373 TI - Severe rhabdomyolysis and acute renal failure due to multiple wasp stings. PMID- 12748377 TI - Vam10p defines a Sec18p-independent step of priming that allows yeast vacuole tethering. AB - YOR068c, termed VAM10 (altered vacuole morphology), lies within the VPS5 gene on the opposite DNA strand. VAM10 deletion causes vacuole fragmentation in vivo. The in vitro fusion of purified yeast vacuoles is stimulated by recombinant Vam10p and blocked by antibody to Vam10p. Vam10p acts early in the priming stage of fusion, independent of Sec18p. After priming, recombinant Vam10p will not stimulate fusion and anti-Vam10p antibodies will not inhibit; Vam10p provides a functional marker for this Sec18p-independent priming step. Pure Vam10p restores normal, Ypt7p-dependent tethering to vacuoles from a vam10Delta strain. PMID- 12748378 TI - Codon bias and frequency-dependent selection on the hemagglutinin epitopes of influenza A virus. AB - Although the surface proteins of human influenza A virus evolve rapidly and continually produce antigenic variants, the internal viral genes acquire mutations very gradually. In this paper, we analyze the sequence evolution of three influenza A genes over the past two decades. We study codon usage as a discriminating signature of gene- and even residue-specific diversifying and purifying selection. Nonrandom codon choice can increase or decrease the effective local substitution rate. We demonstrate that the codons of hemagglutinin, particularly those in the antibody-combining regions, are significantly biased toward substitutional point mutations relative to the codons of other influenza virus genes. We discuss the evolutionary interpretation and implications of these biases for hemagglutinin's antigenic evolution. We also introduce information-theoretic methods that use sequence data to detect regions of recent positive selection and potential protein conformational changes. PMID- 12748379 TI - Trehalose 6-phosphate is indispensable for carbohydrate utilization and growth in Arabidopsis thaliana. AB - Genes for trehalose metabolism are widespread in higher plants. Insight into the physiological role of the trehalose pathway outside of resurrection plant species is lacking. To address this lack of insight, we express Escherichia coli genes for trehalose metabolism in Arabidopsis thaliana, which manipulates trehalose 6 phosphate (T6P) contents in the transgenic plants. Plants expressing otsA [encoding trehalose phosphate synthase (TPS)] accumulate T6P whereas those expressing either otsB [encoding trehalose phosphate phosphatase (TPP)] or treC [encoding trehalose phosphate hydrolase (TPH)] contain low levels of T6P. Expression of treF (encoding trehalase) yields plants with unaltered T6P content and a phenotype not distinguishable from wild type when grown on soil. The marked phenotype obtained of plants accumulating T6P is opposite to that of plants with low T6P levels obtained by expressing either TPP or TPH and consistent with a critical role for T6P in growth and development. Supplied sugar strongly inhibits growth of plants with reduced T6P content and leads to accumulation of respiratory intermediates. Remarkably, sugar improves growth of TPS expressors over wild type, a feat not previously accomplished by manipulation of metabolism. The data indicate that the T6P intermediate of the trehalose pathway controls carbohydrate utilization and thence growth via control of glycolysis in a manner analogous to that in yeast. Furthermore, embryolethal A. thaliana tps1 mutants are rescued by expression of E. coli TPS, but not by supply of trehalose, suggesting that T6P control over primary metabolism is indispensable for development. PMID- 12748380 TI - Immunization with hepatitis C virus-like particles protects mice from recombinant hepatitis C virus-vaccinia infection. AB - We have recently demonstrated that immunization with hepatitis C virus-like particles (HCV-LPs) generated in insect cells can elicit both humoral and cellular immune responses in BALB/c mice. Here, we evaluate the immunogenicity of HCV-LPs in HLA2.1 transgenic (AAD) mice in comparison to DNA immunization. HCV-LP immunization elicited a significantly stronger humoral immune response than DNA immunization. HCV-LP-immunized mice also developed stronger HCV-specific cellular immune responses than DNA-immunized mice as determined by using quantitative enzyme-linked immunospot (ELISpot) assay and intracellular cytokine staining. In BALB/c mice, immunization with HCV-LPs resulted in a >5 log10 reduction in vaccinia titer when challenged with a recombinant vaccinia expressing the HCV structural proteins (vvHCV.S), as compared to 1 log10 decrease in DNA immunization. In HLA2.1 transgenic mice, a 1-2 log10 reduction resulted from HCV LP immunization, whereas no reduction was seen from DNA immunization. Adoptive transfer of lymphocytes from HCV-LP-immunized mice to naive mice provided protection against vvHCV.S challenge, and this transferred immunity can be abrogated by either CD4 or CD8 depletion. Our results suggest that HCV-LPs can induce humoral and cellular immune responses that are protective in a surrogate HCV challenge model and that a strong cellular immunity provided by both CD4 and CD8 effector lymphocytes may be important for protection from HCV infection. PMID- 12748381 TI - Mice lacking methyl-CpG binding protein 1 have deficits in adult neurogenesis and hippocampal function. AB - DNA methylation-mediated epigenetic regulation plays critical roles in regulating mammalian gene expression, but its role in normal brain function is not clear. Methyl-CpG binding protein 1 (MBD1), a member of the methylated DNA-binding protein family, has been shown to bind methylated gene promoters and facilitate transcriptional repression in vitro. Here we report the generation and analysis of MBD1-/- mice. MBD1-/- mice had no detectable developmental defects and appeared healthy throughout life. However, we found that MBD1-/- neural stem cells exhibited reduced neuronal differentiation and increased genomic instability. Furthermore, adult MBD1-/- mice had decreased neurogenesis, impaired spatial learning, and a significant reduction in long-term potentiation in the dentate gyrus of the hippocampus. Our findings indicate that DNA methylation is important in maintaining cellular genomic stability and is crucial for normal neural stem cell and brain functions. PMID- 12748383 TI - Human cathepsin S, but not cathepsin L, degrades efficiently MHC class II associated invariant chain in nonprofessional APCs. AB - MHC class II-restricted antigen presentation plays a central role in the immune response against exogenous antigens. The association of invariant (Ii) chain with MHC class II dimers is required for proper antigen presentation to CD4+ T cells by antigen-presenting cells. MHC class II complexes first traffic through the endocytic pathway to allow Ii chain degradation and antigenic peptide loading before their arrival at the cell surface. In recent years, a considerable effort has been directed toward the identification of proteases responsible for Ii chain degradation. Targeted gene deletion in mice has allowed a precise description of the cysteine proteases involved in the last step of Ii chain degradation. By using nonspecialized cellular models expressing MHC II molecules, we are now exploring the contribution of known cysteine proteases to human Ii chain processing. Surprisingly and contrary to the situation in mouse, cathepsin S was found to be the only human cysteine protease able to efficiently degrade the Ii p10 fragment in epithelial cells. This selectivity has implications for thymic selection and indicates that differences between man and mice are probably more profound at this level than expected. PMID- 12748382 TI - Short- and long-term enhancement of excitatory transmission in the spinal cord dorsal horn by nicotinic acetylcholine receptors. AB - Spinal administration of nicotinic agonists can produce both hyperalgesic and analgesic effects in vivo. The cellular mechanisms underlying these behavioral phenomena are not understood. As a possible explanation for nicotinic hyperalgesia, we tested whether nicotinic acetylcholine receptors (nAChRs) could enhance excitatory transmission onto spinal cord dorsal horn neurons. Whole-cell patch-clamp recordings were performed in neonatal rat spinal cord slices. Activation of nAChRs enhanced glutamatergic synaptic transmission in 59% of dorsal horn neurons tested, and this effect was blocked by methyllycaconitine (10 nM), suggesting a key role for alpha7 nAChRs. Inhibition of acetylcholinesterase with methamidophos also enhanced transmission, demonstrating a similar effect of endogenous acetylcholine. nAChR activation also enhanced transmission by dorsal root entry zone stimulation, suggesting that alpha7 nAChRs on the central terminals of DRG afferents mediate this effect. Paired pre- and postsynaptic stimulation induced long-term potentiation of excitatory inputs to some of the dorsal horn neurons. Long-term potentiation induction was much more prevalent when nicotine was applied during stimulation. This effect also depended on both alpha7 nAChRs and N-methyl-d-aspartate glutamate receptors. Our findings demonstrate that alpha7 nAChRs can contribute to both short- and long-term enhancement of glutamatergic synaptic transmission in the spinal cord dorsal horn and provide a possible mechanism for nicotinic hyperalgesia. PMID- 12748384 TI - Targeted mutation of the outer membrane protein P66 disrupts attachment of the Lyme disease agent, Borrelia burgdorferi, to integrin alphavbeta3. AB - Borrelia burgdorferi, the agent of Lyme disease, expresses several adhesion molecules that are probably required for initial establishment of infection in mammalian hosts, and for colonization of various tissues within the host. The B. burgdorferi outer membrane protein P66 was previously identified as a ligand for beta3-chain integrins by using a variety of biochemical approaches. Although the earlier data suggested that P66 is an adhesin that mediates B. burgdorferi attachment to beta3-chain integrins, lack of genetic systems in B. burgdorferi precluded definitive demonstration of a role for P66 in beta3 integrin attachment by intact borreliae. Recent advances in the genetic manipulation of B. burgdorferi have now made possible the targeted disruption of the p66 gene. Mutants in p66 show dramatically reduced attachment to integrin alphavbeta3. This is, to our knowledge, the first description of the targeted disruption of a candidate B. burgdorferi virulence factor with a known biochemical function that can be quantified, and demonstrates the importance of B. burgdorferi P66 in the attachment of this pathogenic spirochete to a human cell-surface receptor. PMID- 12748386 TI - Crosstalk between cytosolic and plastidial pathways of isoprenoid biosynthesis in Arabidopsis thaliana. AB - In plants, the formation of isopentenyl diphosphate and dimethylallyl diphosphate, the central intermediates in the biosynthesis of isoprenoids, is compartmentalized: the mevalonate (MVA) pathway, which is localized to the cytosol, is responsible for the synthesis of sterols, certain sesquiterpenes, and the side chain of ubiquinone; in contrast, the recently discovered MVA independent pathway, which operates in plastids, is involved in providing the precursors for monoterpenes, certain sesquiterpenes, diterpenes, carotenoids, and the side chains of chlorophylls and plastoquinone. Specific inhibitors of the MVA pathway (lovastatin) and the MVA-independent pathway (fosmidomycin) were used to perturb biosynthetic flux in Arabidopsis thaliana seedlings. The interaction between both pathways was studied at the transcriptional level by using GeneChip (Affymetrix) microarrays and at the metabolite level by assaying chlorophylls, carotenoids, and sterols. Treatment of seedlings with lovastatin resulted in a transient decrease in sterol levels and a transient increase in carotenoid as well as chlorophyll levels. After the initial drop, sterol amounts in lovastatin treated seedlings recovered to levels above controls. As a response to fosmidomycin treatment, a transient increase in sterol levels was observed, whereas chlorophyll and carotenoid amounts decreased dramatically when compared with controls. At 96 h after fosmidomycin addition, the levels of all metabolites assayed (sterols, chlorophylls, and carotenoids) were substantially lower than in controls. Interestingly, these inhibitor-mediated changes were not reflected in altered gene expression levels of the genes involved in sterol, chlorophyll, and carotenoid metabolism. The lack of correlation between gene expression patterns and the accumulation of isoprenoid metabolites indicates that posttranscriptional processes may play an important role in regulating flux through isoprenoid metabolic pathways. PMID- 12748387 TI - Dictyostelium and Acanthamoeba myosin II assembly domains go to the cleavage furrow of Dictyostelium myosin II-null cells. AB - How myosin II localizes to the cleavage furrow of dividing cells is largely unknown. We show here that a 283-residue protein, assembly domain (AD)1, corresponding to the AD in the tail of Dictyostelium myosin II assembles into bundles of long tubules when expressed in myosin II-null cells and localizes to the cleavage furrow of dividing cells. AD1 mutants that do not polymerize in vitro do not go to the cleavage furrow in vivo. An assembly-competent polypeptide corresponding to the C-terminal 256 residues of Acanthamoeba myosin II also goes to the cleavage furrow of Dictyostelium myosin II-null cells. When overexpressed in wild-type cells, AD1 colocalizes with endogenous myosin II (possibly as a copolymer) in interphase, motile, and dividing cells and under caps of Con A receptors but has no effect on myosin II-dependent functions. These results suggest that neither a specific sequence, other than that required for polymerization, nor interaction with other proteins is required for localization of myosin II to the cleavage furrow. PMID- 12748385 TI - An EP2 receptor-selective prostaglandin E2 agonist induces bone healing. AB - The morbidity and mortality associated with impaired/delayed fracture healing remain high. Our objective was to identify a small nonpeptidyl molecule with the ability to promote fracture healing and prevent malunions. Prostaglandin E2 (PGE2) causes significant increases in bone mass and bone strength when administered systemically or locally to the skeleton. However, due to side effects, PGE2 is an unacceptable therapeutic option for fracture healing. PGE2 mediates its tissue-specific pharmacological activity via four different G protein-coupled receptor subtypes, EP1, -2, -3, and -4. The anabolic action of PGE2 in bone has been linked to an elevated level of cAMP, thereby implicating the EP2 and/or EP4 receptor subtypes in bone formation. We identified an EP2 selective agonist, CP-533,536, which has the ability to heal canine long bone segmental and fracture model defects without the objectionable side effects of PGE2, suggesting that the EP2 receptor subtype is a major contributor to PGE2's local bone anabolic activity. The potent bone anabolic activity of CP-533,536 offers a therapeutic alternative for the treatment of fractures and bone defects in patients. PMID- 12748391 TI - The automation of a commercial Fourier transform mass spectrometer to provide a quick and robust method for determining exact mass for the synthetic chemist. AB - Automation of a commercially available Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometer for the routine analysis of the synthetic products from high-speed chemistry is described. The automation includes software written by the instrument manufacturer and in-house developed software; allowing electronic submission of samples from the chemist and e-mailing of results back to the chemist. The use of samples of relatively high concentration (ca 1 mg x mL(-1)) is possible due to the protocol that has been developed, which includes dilution by the autosampler during sample injection. Though high concentrations are used for speed and convenience the amount of sample consumed is still small ca 15 microg per injection. The results from this method have been shown to be both accurate (average error +/- 0.91 ppm) and precise (-0.70 ppm to 2.26 ppm). The system is capable of analysing up to 800 samples per 24 hours. As high speed chemistry becomes more highly utilised within discovery the number of samples requiring accurate mass analysis will rise, and the method we have described will prevent high resolution mass spectrometry becoming the bottleneck in new chemical entity production. The accuracy and precision demonstrated by this method allows high confidence levels in assigned molecular formulae for expected compounds and reduces the number of possible formulae to consider when working with a compound that is not the desired product of a given reaction. PMID- 12748389 TI - Positional cloning of the young mutation identifies an essential role for the Brahma chromatin remodeling complex in mediating retinal cell differentiation. AB - Zebrafish with the young (yng) mutation show a defect in retinal cell differentiation. Here we demonstrate that a mutation in a brahma-related gene (brg1) is responsible for the yng phenotype. Brahma homologues function as essential subunits for SWI/SNF-type chromatin remodeling complexes. Our analysis indicates that brg1 is required for the wave of mitogen-activated protein kinase activity that precedes retinal cell differentiation. Using specific inhibitors of the mitogen-activated protein kinase pathway we show this signal has a direct role in retinal cell differentiation. Lastly, through investigations of mutants in other chromatin remodeling subunits, we provide genetic evidence for gene and tissue specificity of the Brahma chromatin remodeling complex. PMID- 12748388 TI - Mice lacking dipeptidyl peptidase IV are protected against obesity and insulin resistance. AB - Dipeptidyl peptidase IV (DP-IV), a member of the prolyl oligopeptidase family of peptidases, is involved in the metabolic inactivation of a glucose-dependent insulinotropic hormone, glucagon-like peptide 1 (GLP-1), and other incretin hormones. Here, we investigated the impact of DP-IV deficiency on body weight control and insulin sensitivity in mice. Whereas WT mice displayed accelerated weight gain and hyperinsulinemia when fed a high-fat diet (HFD), mice lacking the gene encoding DP-IV (DP-IV-/-) are refractory to the development of obesity and hyperinsulinemia. Pair-feeding and indirect calorimetry studies indicate that reduced food intake and increased energy expenditure accounted for the resistance to HFD-induced obesity in the DP-IV-/- mice. Ablation of DP-IV also is associated with elevated GLP-1 levels and improved metabolic control in these animals, resulting in improved insulin sensitivity, reduced pancreatic islet hypertrophy, and protection against streptozotocin-induced loss of beta cell mass and hyperglycemia. Together, these observations suggest that chronic deletion of DP IV gene has significant impact on body weight control and energy homeostasis, providing validation of DP-IV inhibition as a viable therapeutic option for the treatment of metabolic disorders related to diabetes and obesity. PMID- 12748393 TI - Analysis of polypropyleneglycols using electrospray ionization mass spectrometry. Effects of cationizing agents on the mass spectra. AB - In electrospray ionization mass spectrometry (ESI-MS) of polypropyleneglycol (PPG), effects of cationizing agents were examined. When NaI was used as a cationizing agent, the distribution of multiply-charged ions in the spectra was greatly affected by the ratio of cationizing agent and PPG. However, the distribution was not affected by the use of CH(3)COONH(4). With an increase of cone voltage, fragmentation occurred by in-source collision-induced dissociation (CID) when CH(3)COONH(4) was used. On the contrary, no decomposition of the PPG backbone was observed with NaI. Instead, the intensity of the lower-charged ions, whose mass-to-charge (m/z) ratios are larger, increased because of the elimination of Na(+) with increase of cone voltage. Under optimum conditions for ESI-MS analysis, PPGs that have different molecular weights, different initiators or end groups were easily and accurately characterized. A tandem mass spectrometry (MS/MS) study of NH(4)(+) adduct ions of PPG indicated that a vinyl terminated linear structure is formed at the end group during the fragmentation. PMID- 12748394 TI - Profiling of cyclic hexadepsipeptides roseotoxins synthesized in vitro and in vivo: a combined tandem mass spectrometry and quantum chemical study. AB - High-performance liquid chromatography and tandem mass spectrometry (HPLC/MS/MS) was used for the detection of cyclic hexadepsipeptides roseotoxins produced by Trichothecium roseum. Roseotoxins were found in both submerged standard cultivation on CzapekDox medium and in vivo cultivation extract obtained from an apple. Roseotoxin chromatographic profiles from these two experiments were compared. Product-ion collision-induced dissociation (CID) spectra obtained on an ion trap (electrospray ionisation, ESI) were used for the identification of natural roseotoxins A, B, C and of minor destruxins A and B. The dissociation behavior of roseotoxins is discussed in terms of a fragmentation scheme proposed for describing the dissociation pathways of cyclic peptides. This scheme involves opening of the cyclopeptide ring via formation of oxazolone derivatives and fragmentation of the resulting linear species, which have a free N-terminus and an oxazolone ring at the C-terminus. Some aspects of this fragmentation scheme are underlined by modeling the dissociation channels of roseotoxin A using quantum chemical calculations. The structures of roseotoxin A and destruxin B were verified by nuclear magnetic resonance (NMR) spectroscopy. Structures of three new minor natural roseotoxins [Val(4)]RosA, [MeLxx(4)]RosA and [MeLxx(4)]RosB were deduced by ion cyclotron resonance Fourier transform mass spectrometry (ICR-FT-MS) and ion trap tandem mass spectrometry by examining the pre-separated roseotoxin fraction. PMID- 12748395 TI - Sustained off-resonance irradiation collision-induced dissociation of linear, substituted and cyclic polyesters using a 9.4 T Fourier transform ion cyclotron resonance mass spectrometer. AB - The fragment ions obtained from sustained off-resonance irradiation collision induced dissociation of linear polyesters, substituted polyesters and cyclic polyesters have been characterized using a 9.4 T Fourier transform ion cyclotron resonance mass spectrometer. Charge-induced and charge-remote fragmentation channels, together with the participation of other nucleophilic groups, are proposed for the substituted polyesters. The linear polyesters were found to fragment at equivalent positions along the polymer chain whereas, under the experimental conditions employed, the cyclic polyester produced a single fragment. PMID- 12748392 TI - Effects of internal hydrogen bonds between amide groups: protonation of alicyclic diamides. AB - The proton affinity (PA) of cyclopentane carboxamide 1, cyclohexane carboxamide 2 and their secondary and tertiary amide derivatives S1, S2, T1 and T2, was determined by the thermokinetic method and the kinetic method [PA(1) = 888 +/- 5 kJ mol(1); PA(2) = 892 +/- 5 kJ mol(1); PA(S1) = 920 +/- 6 kJ mol(1); PA(S2) = 920 +/- 6 kJ mol(1); PA(T1) = 938 +/- 6 kJ mol(1); PA(T2) = 938 +/- 6 kJ mol(1)]. Special entropy effects are not observed. Additionally, the effects of protonation have been studied using an advanced kinetic method for all isomers 37 of cyclopentane dicarboxamides and cyclohexane dicarboxamides (with the exception of cis-cyclopentane-1,2-dicarboxamide) and their bis-tertiary derivatives T3T7 by estimating the PA and the apparent entropy of protonation Delta(DeltaS(app)). Finally, the study was extended to bicyclo[2.2.1]hepta-2,5-diene-2,3 dicarboxamide 8 and its bis-tertiary derivative T8, to all stereoisomers of bicyclo[2.2.1]heptane-2,3-dicarboxamide 9, their secondary and tertiary amide derivatives S9 and T9, and to endoendobicyclo[2.2.1]heptane-2,5-dicarboxamide 10 and the corresponding secondary and tertiary derivatives S10 and T10. Compared with 1 and 2, all alicyclic diamides exhibit a significant increase of the PA (DeltaPA) and special entropy effects on protonation. For alicyclic diamides, which can not accommodate a conformation appropriate for building a proton bridge, the values of DeltaPA and Delta(DeltaS(app)) are small to moderate. This is explained by ion / dipole interactions between the protonated and neutral amide group which stabilize the protonated species but hinder the free rotation of the amide groups. If any of the conformations of the alicyclic diamide allows formation of a proton bridge, DeltaPA and Delta(DeltaS(app)) increase considerably. A spectacular case is cis-cyclohexane-1,4-dicarboxamide 7c which is the most basic monocyclic diamide, although generation of the proton bridge requires the unfavorable boat conformation with both amide substituents at a flagpole position. A pre-orientation of the two amide groups in such a 1,4 position in 10 results in a particularly large PA of < 1000 kJ mol(1). The observation of comparable values for Delta(DeltaS(app)) for linear and monocyclic diamides indicates that a major part of the entropy effects originates from freezing the free rotation of the amide groups by formation of the proton bridge. This is corroborated by observing corresponding effects during the protonation of dicarboxamides containing the rigid bicyclo[2.2.1]heptane carbon skeleton, where the only internal movements of the molecules corresponds to rotation of the amide substituents. PMID- 12748390 TI - The receptor kinases LePRK1 and LePRK2 associate in pollen and when expressed in yeast, but dissociate in the presence of style extract. AB - After pollen grains germinate on the stigma, pollen tubes traverse the extracellular matrix of the style on their way to the ovules. We previously characterized two pollen-specific, receptor-like kinases, LePRK1 and LePRK2, from tomato (Lycopersicon esculentum). Their structure and immunolocalization pattern and the specific dephosphorylation of LePRK2 suggested that these kinases might interact with signaling molecules in the style extracellular matrix. Here, we show that LePRK1 and LePRK2 can be coimmunoprecipitated from pollen or when expressed together in yeast. In yeast, their association requires LePRK2 kinase activity. In pollen, LePRK1 and LePRK2 are found in an approximately 400-kDa protein complex that persists on pollen germination, but this complex is disrupted when pollen is germinated in vitro in the presence of style extract. In yeast, the addition of style extract also disrupts the interaction between LePRK1 and LePRK2. Fractionation of the style extract reveals that the disruption activity is enriched in the 3- to 10-kDa fraction. A component(s) in this fraction also is responsible for the specific dephosphorylation of LePRK2. The style component(s) that dephosphorylates LePRK2 is likely to be a heat-stable peptide that is present in exudate from the style. The generally accepted model of receptor kinase signaling involves binding of a ligand to extracellular domains of receptor kinases and subsequent activation of the signaling pathway by receptor autophosphorylation. In contrast to this typical scenario, we propose that a putative style ligand transduces the signal in pollen tubes by triggering the specific dephosphorylation of LePRK2, followed by dissociation of the LePRK complex. PMID- 12748396 TI - Atmospheric pressure electrospray ionization mass spectra of glucose and 2 fluorodeoxyglucose for quantitative analysis of 2-fluorodeoxyglucose by high performance liquid chromatography. AB - 2-Fluoro-2-deoxy-D-glucose (FDG) labeled by fluorine-18 is the most widely used radiopharmaceutical for positron emission tomography (PET). For high-performance liquid chromatography (HPLC)/MS assay and quality control, the mass spectra of FDG and glucose (Glc) in organic + water solutions were studied by flow injection analysis (FIA) and in a chromatographic eluate. In acetonitrile (MeCN) + 0.025% ammonium formate (NH(4)HCO(2)) solvent (80 : 20), electrospray ionisation (ESI) of glucose-FDG provides M.NH(4)(+) and 2M.Na(+) (M = Glc or FDG) as the most intense positive ions. Formation of the latter ions and also of M.MeCN.Na(+) and 2MeCN. Na(+) is typical of the presence of NaCl in the ESI inlet. The positive ions include heavier ions corresponding to the impurities separated by HPLC and also to the cross-ring fragmentation of complexes (2FDG. aMeCNX)L, where a = 0 or 1, L is either Na(+) or NH(4)(+) and X is a fragmented pyranose or anhydropyranose residue. The second most abundant Glc negative ion is m/z = 359 which was interpreted as (2GlcH(+))(). The negative-ion spectrum of FDG has dominating lines due to FDG.HCO(2)() ions at m/z 227 and also (2FDGH(+))() at m/z 363. The m/z 363 signal is suppressed in the presence of NaCl at a molar ratio of 4 : 1 to NH(4)HCO(2), while the ions at m/z 217 and 219, i.e. FDG.Cl(), become three times more intense than FDG.HCO(2)(). The latter ion appears to be most suitable as an analytical signal for chemical analysis of FDG at m/z 226 and 227. Limits of FDG quantitation (LOQ) of 19 ng and 21 ng were found for the 200(+) and 227() ion signals, respectively, and are wholly adequate for verification of total FDG content in radiopharmaceuticals. PMID- 12748397 TI - Towards the standard-module approach to disulfide-linked polypeptide nanostructures. I. Methodological prerequisites and mass spectrometric characterization of the test two-loop structure. AB - Potentially biologically-active nanostructures can be created from single chains of unmodified peptides by cross-linking different regions of the chain by disulfide bonds and cleaving the chain at specified sites to obtain the final configuration. The availability of techniques for assembly and characterization of such structures was tested on a two-loop structure created from a 21-residue linear peptide. Directed intra-molecular disulfide bond formation was performed by inserting partial sequences favoring intra-molecular SS bond formation ("loops") separated by partial sequences disfavoring such a process ("spacers") into the precursor sequence. Peptide bond cleavage by partial acid hydrolysis at specific sites (GG, NP/DP) inside the loops opened them; the same process in the spacer separated the loops. Synthesis, oxidation and bond cleavage were monitored by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI ToF MS). The hydrolysis fragments of the produced nanostructures were characterized by tandem electrospray ionization Fourier transform mass spectrometry (ESI FT-MS) with collisional and electron capture dissociations. The latter technique was especially useful as it cleaves SS bonds preferentially. The feasibility of the proposed synthesis approach and the adequacy of the analysis techniques for the test structure were demonstrated. PMID- 12748398 TI - Derivatization in mass spectrometry--1. Silylation. AB - This is the first of a series of reviews on the application of derivatization in mass spectrometry. A description is given of advances in silylation as a powerful tool used for increasing the volatility, thermal and thermo-catalytic stability, and chromatographic mobility of polar and unstable organic compounds. In addition to chemical aspects of silylation, mass spectral properties of silyl derivatives useful for structure determination and quantitation of various organic and biologically-active compounds, mainly by GC/MS, are described. Practically all tested and widely used silylating agents are described. The role of comprehensive libraries containing reference mass spectra for various silyl derivatives and search systems in structure determination is emphasized. Applications of silylation for particular analyses are summarised. PMID- 12748399 TI - Mass-selected resonance-enhanced multiphoton ionisation spectra of laser-desorbed molecules for environmental analysis: 16 representative polycyclic aromatic compounds. AB - Mass-selected gas-phase UV spectra of laser-desorbed molecules at room temperature have been measured via resonance-enhanced multiphoton ionisation (REMPI) and time-of-flight mass selection. The wavelength range of 260 to 320 nm is optimal for detection of polycyclic aromatic hydrocarbons (PAHs) using REMPI mass spectrometry. A new laser desorption/laser ionisation source has been used which features a compact size and thermal equilibrium of the desorbed molecules. 16 PAHs have been investigated which have been selected by the US Environmental Protection Agency (EPA). These 16 EPA-PAHs are commonly used world-wide to characterise the PAH-load of environmental samples. PMID- 12748400 TI - Atmospheric pressure matrix-assisted laser desorption/ionisation ion trap mass spectrometry of synthetic polymers: a comparison with vacuum matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. AB - Atmospheric pressure matrix-assisted laser desorption/ionisation quadrupole ion trap (AP-MALDI/QIT) mass spectrometry has been investigated for the analysis of polyethylene glycol (PEG 1500) and a hyperbranched polymer (polyglycidol) in the presence of alkali-metal salts. Mass spectra of PEG 1500 obtained at atmospheric pressure showed dimetallated matrix/analyte adducts, in addition to the expected alkali-metal/PEG ions, for all matrix/alkali-metal salt combinations. The relative intensities of the desorbed ions were dependent on the matrix, the alkali-metal salt added to aid cationisation and the ion trap interface conditions [capillary temperature, in-source collisionally-induced dissociation (CID)]. These data indicate that the adducts are rapidly stabilised by collisional cooling enabling them to be transferred into the ion trap. Experiments using identical sample preparation conditions were carried out on a vacuum MALDI time-of-flight (ToF) mass spectrometer. In all cases, vacuum MALDI ToF spectra showed only alkali-metal/PEG ions and no matrix/analyte adducts. The tandem mass spectrometry (MS/MS) capability of the ion trap has been demonstrated for a lithiated polyglycol yielding a rich fragment-ion spectrum. Analysis of the hyperbranched polymer polyglycidol by AP-MALDI/QIT reveals the characteristic ion series for these polymers as also observed under vacuum MALDI-ToF conditions. PMID- 12748402 TI - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of dextran and dextrin derivatives. AB - Application of matrix-assisted laser desorption/ionization (MALDI) to the analysis of dextran and dextrin derivatives, specifically glucose saccharides, by time-of-flight (TOF) mass spectrometry has been reported. MALDI-TOF analysis was carried out on alpha-, beta- and gamma-cyclodextrin, two O-methylated-beta cyclodextrins of differing degrees of substitution (DS) and dextrans (a linear glucose saccharide), as pure and doped solutions and as mixtures of two or more of these. Doping was carried out with trace amounts of inorganic salts. The purpose of the analysis of the cyclodextrins was to determine whether they would form inclusion complexes with the various cations added, or whether less specific cation addition/exchange was occurring either prior to desorption or in the gas phase. PMID- 12748401 TI - Structural characterisation by both positive- and negative-ion electrospray mass spectrometry of partially methyl-esterified oligogalacturonides purified by semi preparative high-performance anion-exchange chromatography. AB - The off-line coupling of high-performance anion-exchange chromatography (HPAEC) to electrospray ionisation/ion trap mass spectrometry (ESI-ITMS) is described. The Dionex carbohydrate membrane desalter (CMD) has been assessed as an on-line chromatographic desalting system to remove the high sodium concentration necessary for the HPAEC separation of partially methyl-esterified oligogalacturonides. The developed HPAEC configuration proved to be suitable for indirect coupling with ESI-ITMS. This paper provides some interesting features of positive- and negative-ion multistage tandem mass spectrometry (MS(n)) analysis of these acidic oligosaccharides. The spectra acquired in both negative- and positive-ion modes show characteristic fragment ions resulting from glycosidic bond and cross-ring cleavages. Some new mass spectrometric fragmentation routes are also described. The positive-ion mode gave more complex spectra but was as informative as the negative-ion mode. ESI-ITMS was revealed to be, as previously reported from direct use on an unseparated enzymatic digest, a powerful sequencing technique for the determination of linkage type and the methyl ester distribution of partially methyl-esterified oligogalacturonides. Moreover, unlike matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-ToF MS), it gives valuable information on the elution behaviour of these oligomers in relation to their structure, namely the HPAEC co-elution of isomeric structures. PMID- 12748403 TI - Maximum information approach to scale description for affective measures based on the Rasch model. AB - Using the Rasch model for ordered categories, this paper provides a method for qualitative interpretations of data from an affective measure such as an attitude scale or survey instrument. The Bock procedure is first used to partition the total item information to each response category. The location of each response category is determined by maximizing the information associated with this category. The response categories that cluster around a given point on the latent trait are then used to provide a qualitative description of this point. The description is explicit in terms of the behaviors or activities that are likely to be displayed by a respondent at this point. An illustration is provided using an alumni survey used at a large university in the southeast. PMID- 12748404 TI - Measuring client satisfaction with public education I: meeting competing demands in establishing state-wide benchmarks. AB - By its very nature, a large-scale evaluation of client satisfaction with public education using a quantitative approach, places almost impossibly competing demands on the research methodology. This paper reports on the use of a suite of Rasch measurement techniques to meet the competing demands in establishing state wide benchmarks relating to the School Opinion Survey carried out over 1200 government schools in one state of Australia. Although the evaluation had to establish system-wide representative parent and student benchmarks, meaningful quantitative estimates of client satisfaction had to be provided at the smallest public schools. The final 20-item School Opinion Survey Parent and Student Forms were designed following feedback from the administration of trial forms. Instrument development was monitored by the results of Rasch modeling. The Rasch modeling property of specific objectivity was empirically verified when calculation of identical benchmark estimates resulted from the construction of simulated population proportional samples using sample:population size weightings. PMID- 12748405 TI - Developing an initial physical function item bank from existing sources. AB - The objective of this article is to illustrate incremental item banking using health-related quality of life data collected from two samples of patients receiving cancer treatment. The kinds of decisions one faces in establishing an item bank for computerized adaptive testing are also illustrated. Pre-calibration procedures include: identifying common items across databases; creating a new database with data from each pool; reverse-scoring "negative" items; identifying rating scales used in items; identifying pivot points in each rating scale; pivot anchoring items at comparable rating scale categories; and identifying items in each instrument that measure the construct of interest. A series of calibrations were conducted in which a small proportion of new items were added to the common core and misfitting items were identified and deleted until an initial item bank has been developed. PMID- 12748406 TI - Breakthrough measuring neighborhoods. AB - An empirical strategy is presented for transforming ordinal counts and percentages to interval scale measures by recoding them as ordered categories and estimating Rasch model rating scale parameters. This strategy is demonstrated for a neighborhood construct socioeconomic disadvantage operationally defined by eight characteristics of Chicago neighborhoods (N = 77). Results show surprisingly sound model fit and satisfactory scale invariance between 1980 and 1990 census. A striking finding obscured by traditional methods is many Chicago neighborhoods are four times more disadvantaged than official U.S. poverty threshold. Intramodel construct validation confirms this scale structure is consistent with sociological expectations about property values, income, and race. A general benefit of this approach over conventional categorical socioeconomic indices is neighborhood measurement on a linear scale. PMID- 12748407 TI - Rasch fit statistics as a test of the invariance of item parameter estimates. AB - The invariance of the estimated parameters across variation in the incidental parameters of a sample is one of the most important properties of Rasch measurement models. This is the property that allows the equating of test forms and the use of computer adaptive testing. It necessarily follows that in Rasch models if the data fit the model, than the estimation of the parameter of interest must be invariant across sub-samples of the items or persons. This study investigates the degree to which the INFIT and OUTFIT item fit statistics in WINSTEPS detect violations of the invariance property of Rasch measurement models. The test in this study is a 80 item multiple-choice test used to assess mathematics competency. The WINSTEPS analysis of the dichotomous results, based on a sample of 2000 from a very large number of students who took the exam, indicated that only 7 of the 80 items misfit using the 1.3 mean square criteria advocated by Linacre and Wright. Subsequent calibration of separate samples of 1,000 students from the upper and lower third of the person raw score distribution, followed by a t-test comparison of the item calibrations, indicated that the item difficulties for 60 of the 80 items were more than 2 standard errors apart. The separate calibration t-values ranged from +21.00 to -7.00 with the t-test value of 41 of the 80 comparisons either larger than +5 or smaller than -5. Clearly these data do not exhibit the invariance of the item parameters expected if the data fit the model. Yet the INFIT and OUTFIT mean squares are completely insensitive to the lack of invariance in the item parameters. If the OUTFIT ZSTD from WINSTEPS was used with a critical value of | t | > 2.0, then 56 of the 60 items identified by the separate calibration t-test would be identified as misfitting. A fourth measure of misfit, the between ability-group item fit statistic identified 69 items as misfitting when a critical value of t > 2.0 was used. Clearly relying solely on the INFIT and OUTFIT mean squares in WINSETPS to assess the fit of the data to the model would cause one to miss one of the most important threats to the usefulness of the measurement model. PMID- 12748408 TI - Measuring attitudes and behaviors to studying and learning for university students: a Rasch measurement model analysis. AB - A Studying and Learning Scale was created using a model of Motivation (sets of ordered stem-items based on Striving for Excellence, Desire to Learn and Personal Incentives), with each item answered from three self-reported perspectives (an Ideal Self-view, a Capability Self-view, and a Studying and Learning Self-view). The response categories were the number of subjects studied. The stem-item sample was 23, each answered in three aspects, so each stem-item had three 'difficulties', making an effective item sample of 69. The person convenience sample was 372 students in education at an Australian university. The 69 items fit a Rasch measurement model and formed a scale in which the 'difficulties' of the items were ordered from 'easy' to 'hard' and the student measures of Studying and Learning were ordered from 'low' to 'high'. The person separation reliability was high at 0.94. The response categories were answered consistently and logically and the results supported many (but not all) of the conceptually ordered-by-difficulty item patterns. Students found it 'easy' to form a high view of How they would like to be, much 'harder' to form a high view of What they think they are capable of doing and even 'harder' to perform, at a high level, their Studying and Learning behavior for all stem-items, in accordance with the model. PMID- 12748409 TI - Rasch techniques for detecting bias in performance assessments: an example comparing the performance of native and non-native speakers on a test of academic English. AB - The use of common tasks and rating procedures when assessing the communicative skills of students from highly diverse linguistic and cultural backgrounds poses particular measurement challenges, which have thus far received little research attention. If assessment tasks or criteria are found to function differentially for particular subpopulations within a test candidature with the same or a similar level of criterion ability, then the test is open to charges of bias in favour of one or other group. While there have been numerous studies involving dichotomous language test items (see e.g. Chen and Henning, 1985 and more recently Elder, 1996) few studies have considered the issue of bias in relation to performance based tasks which are assessed subjectively, via analytic and holistic rating scales. The paper demonstrates how Rasch analytic procedures can be applied to the investigation of item bias or differential item functioning (DIF) in both dichotomous and scalar items on a test of English for academic purposes. The data were gathered from a pilot English language test administered to a representative sample of undergraduate students (N= 139) enrolled in their first year of study at an English-medium university. The sample included native speakers of English who had completed up to 12 years of secondary schooling in their first language (L1) and immigrant students, mainly from Asian language backgrounds, with varying degrees of prior English language instruction and exposure. The purpose of the test was to diagnose the academic English needs of incoming undergraduates so that additional support could be offered to those deemed at risk of failure in their university study. Some of the tasks included in the assessment procedure involved objectively-scored items (measuring vocabulary knowledge, text-editing skills and reading and listening comprehension) whereas others (i.e. a report and an argumentative writing task) were subjectively-scored. The study models a methodology for estimating bias with both dichotomous and scalar items using the programs Quest (Adams and Khoo, 1993) for the former and ConQuest (Wu, Adams and Wilson, 1998) for the latter. It also offers answers to the practical questions of whether a common set of assessment criteria can, in an academic context such as this one, be meaningfully applied to all subgroups within the candidature and whether analytic criteria are more susceptible to biased ratings than holistic ones. Implications for test fairness and test validity are discussed. PMID- 12748410 TI - Iodine deficiency in vegetarians and vegans. AB - Iodine content in food of plant origin is lower in comparison with that of animal origin due to a low iodine concentration in soil. Urinary iodine excretion was assessed in 15 vegans, 31 lacto- and lacto-ovovegetarians and 35 adults on a mixed diet. Iodine excretion was significantly lower in alternative nutrition groups - 172 microg/l in vegetarians and 78 microg/l in vegans compared to 216 microg/l in subjects on a mixed diet. One fourth of the vegetarians and 80% of the vegans suffer from iodine deficiency (iodine excretion value below 100 microg/l) compared to 9% in the persons on a mixed nutrition. The results show that under conditions of alternative nutrition, there is a higher prevalence of iodine deficiency, which might be a consequence of exclusive or prevailing consumption of food of plant origin, no intake of fish and other sea products, as well as reduced iodine intake in the form of sea salt. PMID- 12748411 TI - Mechanism of increased plasma glucose levels after oral glucose ingestion in normal-weight middle-aged subjects. AB - BACKGROUND/AIMS: To determine the decline in glucose tolerance in normal-weight middle-aged subjects, we performed a cross-sectional study using double-labelled oral glucose tolerance tests in 8 middle-aged (46.3 +/- 0.9 years) and in 8 young (23.6 +/- 0.5) subjects with similar normal body weight. METHODS: Plasma glucose was labelled by an infusion of dideuterated glucose started 120 min before ingestion of 1 g/kg of naturally (13)C-enriched corn starch glucose. Glucose levels, substrate oxidation (indirect calorimetry) and exogenous glucose oxidation ((13)C enrichment of expired CO(2)) were monitored for 330 min. RESULTS: In the middle-aged subjects, the appearance of exogenous glucose was reduced (723 +/- 52 mg/kg/330 min) compared to young subjects (864 +/- 38; p < 0.05), and systemic glucose production was normal. Plasma glucose levels were increased due to a reduced glucose disappearance rate (middle aged: 1,046 +/- 61 mg/kg/330 min vs. young 1,242 +/- 67; p < 0.05), concerning both oxidative and non-oxidative disposal. This reduction was no longer apparent when the results were normalized for fat-free mass. Insulin levels were similar in young and middle-aged subjects. CONCLUSION: In normal-weight middle-aged individuals, glucose intolerance is mainly due to the reduction in the mass of fat-free glucose-utilizing tissues. The higher plasma glucose levels enable normal glucose supply to peripheral tissues, and increase splanchnic glucose uptake. PMID- 12748412 TI - Effects of short-term modified fasting on sleep patterns and daytime vigilance in non-obese subjects: results of a pilot study. AB - BACKGROUND: Periodically repeated short-term fasting is a frequently practised tradition worldwide. Empirical reports suggest that during fasting periods the quality of sleep and daytime performance are improved. The effects of a home based 1-week modified fasting on sleep patterns and daytime vigilance and performance were analysed in 15 healthy non-obese volunteers. METHODS: Sleep was measured by polysomnography before and after a 7-day fasting period; sleep inventories with assessment of daytime performance were collected throughout the observation period. Blood samples and urine were drawn at the beginning and at the end of fasting. RESULTS: 13 subjects (12 females, 1 male; age 41.2 +/- 13.4 years; BMI 23.9 +/- 4.2 kg/m(2)) completed the fasting period; weight decreased from 66.5 +/- 11.7 kg to 63 +/- 11.9 kg. Compared to baseline, a significant decrease in arousals, a decrease in periodic leg movements (PLM) and a non significant increase in REM sleep were observed at the end of fasting. Subjective sleep ratings showed a fasting-induced increase in global quality of sleep, daytime concentration, vigour and emotional balance. Clinical laboratory tests showed a decrease in serum magnesium; urinary melatonin excretion decreased moderately. CONCLUSION: This open pilot study demonstrates that along with a decrease in sleep arousals a 1-week fasting period promotes the quality of sleep and daytime performance in non-obese subjects. The observed decrease in PLM might point to a nutritional modification of brain dopaminergic functions. In terms of evolutionary development, an improved daytime performance during periods of food deprivation could have been beneficial for the success in search for food. PMID- 12748413 TI - Copper supplementation has no effect on markers of DNA damage and liver function in healthy adults (FOODCUE project). AB - BACKGROUND/AIMS: Copper is routinely used in the laboratory to promote oxidation in vitro. However, copper concentrations are million-fold higher than physiological concentrations and, in contrast, accumulating evidence suggests that copper may have an antioxidant role in vivo. The aim of this study was to provide data on how increased intake of copper affected mononuclear leukocyte DNA damage and liver function in healthy young free-living men and women. METHODS: The study design was a double-blind repeated crossover trial with treatment and intervening placebo periods, each of 6 weeks' duration. The following supplementations were given orally in sequence: CuSO(4) at a dose of 3 mg copper/day and copper amino acid chelates at doses of 3 and 6 mg copper/day. Oxidative DNA damage was assessed using a modification of the alkaline Comet assay incorporating an endonuclease III digestion step. The assessment of liver function was by measurement of the liver enzymes, alanine aminotransferase and L gamma-glutamyltransferase. RESULTS: There was no significant alteration in mononuclear leukocyte DNA damage or on liver function after 6 weeks of copper supplementation at two doses (3 and 6 mg/day). CONCLUSIONS: Copper supplementation (giving total copper intake at the highest level of 7 mg/day) did not induce DNA damage or adversely affect liver function in healthy adults. PMID- 12748414 TI - Effects of a dietary thermally oxidized fat on thyroid morphology and mRNA concentrations of thyroidal iodide transporter and thyroid peroxidase in rats. AB - BACKGROUND/AIM: Recent studies demonstrated that feeding oxidized fats increases the concentrations of total and free thyroxine in blood of rats and pigs. This finding suggested that oxidized fats affect the function of the thyroid gland. This study investigates the effects of a thermally oxidized dietary fat on the morphology of the thyroid gland and on the expression of proteins (Na(+)/I(-) symporter, thyroid peroxidase) involved in the synthesis of thyroid hormones in rats at different dietary iodine concentrations. METHODS: An experiment was conducted with 48 growing male Sprague-Dawley rats which were allotted to four groups of 12 animals each. According to a bifactorial experimental design, the rats received semisynthetic diets with 10% of either a fresh or an oxidized fat, with low (50 microg/kg) or adequate (400 microg/kg) iodine concentrations, over a period of 38 days. The oxidized fat was prepared by heating sunflower oil at a temperature of 55 degrees C for a period of 5 weeks. The oxidized fat had much higher concentrations of lipid peroxidation products than the fresh fat as assessed by determining the peroxide concentrations (877 vs. 33 mEq O(2)/kg) and those of thiobarbituric acid reactive substances (25 vs. 0.7 micromol/kg). RESULTS: Rats fed the diets containing oxidized fat had higher concentrations of total and free thyroxine in plasma, a greater height of thyroid epithelial cells, a smaller diameter of thyroid follicle lumen, a lower Na(+)/I(-) symporter mRNA concentration, and a higher thyroid peroxidase mRNA concentration than rats fed the fresh fat (p < 0.05 for all effects). The concentrations of triiodothyronine and thyrotropin were not different between rats fed the fresh fat and those fed the oxidized fat. The dietary iodine supply also had significant effects on some of the parameters analyzed. There were no interactions between type of fat and dietary iodine concentrations. CONCLUSION: The rat model used here shows that dietary oxidized fats affect the morphology and the function of the thyroid gland, irrespective of the dietary iodine supply. PMID- 12748415 TI - Blood pressure and vitamin C and fruit and vegetable intake. AB - BACKGROUND: A high vitamin C intake was recently associated with lower blood pressure levels. We examined this association and compared it with that between blood pressure and fruit and vegetable intake among German adults. METHODS: Complete data were available for 1,628 women and 1,340 men, aged 18-79 years, who participated in the German Nutrition Survey, a subsample of the National Health Survey 1998. The participants completed a health and lifestyle questionnaire and underwent a medical examination and a comprehensive dietary interview. Multiple regression models were used to analyze the association between blood pressure and vitamin C and fruit and vegetable intakes. The associations were adjusted for age and body mass index and in addition for smoking among women and alcohol intake and sport activity among men. RESULTS: Systolic blood pressure showed a significant inverse association with fruit and vegetable but not with vitamin C intake among women. If information about vitamin C and fruit and vegetable intakes was considered simultaneously, a high fruit and vegetable intake was stronger associated with lower systolic blood pressure levels as compared with a high vitamin C intake among women. Among men, we did not observe significant associations between blood pressure and vitamin C and fruit and vegetable intakes. CONCLUSION: The total vitamin C intake seems to be less associated with blood pressure levels as compared with the intake of fruit and vegetables among women. PMID- 12748416 TI - Use of olive oil based emulsions as an alternative to soybean oil based emulsions in total parenteral nutrition and their effects on liver regeneration following hepatic resection in rats. AB - BACKGROUND/AIM: The main objective of this study was to compare the effects of olive oil to those of soybean oil on liver tissue regeneration following hepatic resection in rats. METHODS: Seventy albino Wistar rats were randomly assigned to seven groups which contained 10 rats each. Group 1 was the sham-treated group; groups 2 and 3 received total parenteral nutrition (TPN) containing soybean oil lipid emulsions (20% Lipofundin MCT/LCT) for 48 or 72 h; groups 4 and 5 received TPN containing olive oil (80%)/soybean oil (20%) lipid emulsions (ClinOleic 20%) for 48 or 72 h; group 6 was the control group for 48 h, and group 7 was the control group for 72 h. TPN was given via internal jugular vein, and 70% hepatic resection was performed in the study groups. In addition, hepatic resections with no TPN were performed in the control groups, except the sham group. Relative liver weight, mitotic index, proliferating cell nuclear antigen labeling index, and carnitine levels in liver tissue samples were used to assess hepatic regeneration. Thiobarbituric acid reactive substances were measured as an index of lipid peroxidation and oxidative tissue damage. Alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase were parameters for the liver function. RESULTS: The relative liver weight increased significantly with minimal hepatosteatosis in the olive oil treated groups. Lipid peroxidation significantly decreased with near-normal serum levels of the liver function parameters in the olive oil/soybean oil treated groups, and mitotic index and proliferating cell nuclear antigen labeling index reached their maximum level in rats receiving TPN containing any kind of lipid emulsions for 48 h following resection. CONCLUSIONS: The use of olive oil/soybean oil lipid emulsions in TPN has important beneficial effects on the liver regeneration, and the antioxidant properties of olive oil originating from its natural components indirectly contribute to the liver regeneration in rats. PMID- 12748418 TI - Severe acute pancreatitis: when to be concerned? AB - The fate of a patient with acute pancreatitis largely depends on early recognition of the severity of the disease. Acute pancreatitis is severe when organ failure and/or pancreatic necrosis occur. Whereas simple and low cost parameters are available for the detection of organ failure, the detection and extent of pancreatic necrosis requires a costly contrast-enhanced computed tomography. This is not always available in all institutions. This review article which considers when the clinician should be concerned about his patient diagnosed with acute pancreatitis, summarizes possibilities for diagnosing clinical severe (that is organ failure) and radiological severe (that is the necrotizing form of the disease) pancreatitis using simple and inexpensive parameters which are available everywhere. At present, a CT scan should be performed in those patients with alcohol etiology, those admitted to hospital early (time interval between the onset of symptoms and admission to hospital of less than 24 h, those presenting with rebound tenderness and/or guarding, a lipase >1,000 U/l, as well as a raised hematocrit and blood glucose. The evaluation, furthermore, shows that some parameters have a sufficiently high negative predictive value of >90%, which may render a contrast-enhanced CT scan unnecessary in the early stages, unless the patient fails to improve. These parameters include non-alcohol etiology, time interval between onset and admission to hospital longer than 24 h, no guarding or rebound tenderness on admission, low or normal hematocrit and nonelevated blood glucose. It is necessary to look further on simple low cost and more valid parameters on admission in order to reliably distinguish between necrotizing pancreatitis and interstitial pancreatitis. PMID- 12748417 TI - Angiotensin-converting enzyme inhibition induces death receptor apoptotic pathways in erythroid precursors following renal transplantation. AB - BACKGROUND: Posttransplant erythrocytosis (PTE) is a condition that occurs in kidney transplant patients and is characterized by increase in hematocrit above 51%. While its pathogenesis remains unclear, angiotensin-converting enzyme inhibitors (ACEI) have been used successfully in the treatment of PTE. We have previously shown that ACEI induce apoptosis in the peripheral erythroid precursors from patients with PTE. In the current study we elucidate the molecular mechanisms of ACEI-induced apoptosis. METHODS: Peripheral CD34+ cells were obtained from four normal controls, five normal kidney transplants, and six kidney transplants with PTE, before and after treatment with ACEI. We evaluated the expression of a variety of apoptotic factors by quantitative reverse transcription-multiplex polymerase chain reaction, Western blot and immunocytochemistry. RESULTS: ACEI resulted in a significant induction of Fas, FADD, and TRADD mRNAs in renal transplant patients with or without PTE. No changes were noted in the expression of mRNAs encoding Bcl-2, Bcl-xL, Bax, caspase 8, caspase 3, or GAPDH. ACEI also resulted in a significant upregulation of Fas, FADD and TRADD protein expression, and their localization predominantly at the plasma membrane. CONCLUSIONS: Our results suggest that ACEI therapy induces apoptosis in erythrocyte progenitor cells of renal transplant patients at least in part via induction of death receptor apoptotic cascades. PMID- 12748419 TI - Natural history of organ failure in acute pancreatitis. AB - The majority of deaths from acute pancreatitis are a consequence of multiple organ dysfunction syndrome (MODS). In order to influence the outcome of this condition, we need to have a better understanding of the natural history of acute pancreatitis, particularly when complicated by MODS. This paper reviews the recent literature dealing with the natural history of acute pancreatitis and the relationship between MODS, pancreatic necrosis and mortality. PMID- 12748420 TI - Useful markers for predicting severity and monitoring progression of acute pancreatitis. AB - BACKGROUND: The main problem in staging acute pancreatitis is the lack of accurate predictors of disease severity and of markers for progression of acute pancreatitis. METHODS: We reviewed the literature for all candidate markers of acute pancreatitis and graded their usefulness and practicability for prediction of severe pancreatitis and for monitoring disease progression. RESULTS: Several markers can differentiate mild and severe cases of acute pancreatitis with a high positive predictive value. Trypsinogen activation peptide and procalcitonin show significant differences in patients with mild and severe disease already on admission. While most parameters peak early and decrease rapidly thereafter, C reactive protein (CRP), phospholipase A(2), procalcitonin and serum amyloid A are reliable predictors with persistently elevated levels in severe disease. CRP is still the reference parameter of all predictors indicating severe disease and pancreatic necrosis. So far, no single parameter has been developed which is suitable for early prediction of infected pancreatic necrosis. CONCLUSION: Of all markers available today, CRP is the 'gold standard' in predicting the severity of acute pancreatitis, but procalcitonin seems to be a promising tool to monitor the progression of the disease. CRP has already been established in clinical routine. For procalcitonin, a practicable assay is also available and could easily be adopted into clinical routine. PMID- 12748421 TI - Outcome of open necrosectomy in acute pancreatitis. AB - Twenty percent of all acute pancreatitis patients present with necrotizing pancreatitis. Infected necrosis is responsible for 80% of deaths in the course of the disease though, thanks to antibiotic prophylaxis, the infection rate is decreasing. When infection occurs, the prognosis is poor, and the need for debridement and drainage of the necrosis is mandatory. The aim of this study was to review the most recent literature in order to present an update of open surgical management of infected necrosis. PMID- 12748422 TI - Guiding the reluctant. A primer on guidelines in general and pancreatitis in particular. AB - Within the past 5 years, no fewer than 6 guidelines for the management of acute pancreatitis have appeared in the literature, including the current submission from the International Association of Pancreatology. When these collected guidelines are subjected to comparison, however, marked similarities emerge between the proposals, and significant differences are rare. Surprisingly, neither the passage of time nor the application of evidence-based medicine techniques to the creation of guidelines for acute pancreatitis has resulted in substantive changes to the original guidelines offered by the Atlanta Symposium. Moreover, it is important to realize that, despite claims of objectivity, opinion may enter into the process of guideline creation whenever proposals are based upon lesser levels of evidence. Until the guidelines common to these collected proposals have been supported by randomized controlled trials, it is unlikely that we need any more guidelines for acute pancreatitis. PMID- 12748423 TI - Factors predicting mortality in severe acute pancreatitis. AB - Acute pancreatitis (AP) is a common disorder in which ensuing serious complications may lead to a fatal outcome in patients. BACKGROUND/AIMS: To describe a large series of patients with severe AP (SAP) who were admitted to our hospital and to identify factors predicting mortality. PATIENTS AND METHODS: In a retrospective study, all patients with SAP diagnosed between February 1996 and October 2000 according to the Atlanta criteria were studied. RESULTS: Out of a total of 363 AP patients, 67 developed SAP. The mean age of the patients was 69; the commonest etiology was biliary; 55.2% developed necrosis; the commonest systemic complication was respiratory failure (44.7%), followed by acute renal failure (35.8%) and shock (20.9%). A total of 31.3% of the patients died. Factors significantly related to mortality were age, upper digestive tract bleeding, acute renal failure, respiratory failure and shock by univariate analysis. However, pseudocysts seemed to have a protective effect. By multivariate analysis, independent prognostic factors were age, acute renal failure and respiratory failure. CONCLUSIONS: Patients with SAP mainly died due to systemic complications, especially acute renal failure and respiratory failure. Necrosis (in the absence or presence of infection) was not correlated with increased mortality. A pseudocyst was found to be a protective factor, probably because the definition itself led to the selection of patients who had survived multiorgan failure. PMID- 12748424 TI - A study on the activation peptide released from procarboxypeptidase B (CAPAP) and anionic trypsinogen in patients with acute abdominal disorders of non-pancreatic origin. AB - BACKGROUND: The activation peptide released from procarboxypeptidase B, CAPAP, is a marker of the activation of pancreatic enzymes in acute pancreatitis while anionic trypsinogen (AT) levels in urine relate to leakage of unactivated proenzymes. Data on these markers in patients suffering from severe acute abdominal disorders of non-pancreatic origin are lacking. PURPOSE: To examine levels of CAPAP and AT in serum and urine from patients with severe acute abdominal disorders of non-pancreatic origin in order to better define the diagnostic specificity of these two markers in severe acute pancreatitis in relation to other acute intra-abdominal disorders. SUBJECTS AND METHODS: The study included 54 patients with severe acute abdominal disorders of non pancreatic origin with an APACHE II score >3. Immunoreactive CAPAP (irCAPAP) and immunoreactive AT (irAT) were measured in serum and urine using specific immunoassays. RESULTS: In urine, irCAPAP levels were mildly increased (>2 nmol/l) in 13% of the patients with severe acute abdominal diseases of non-pancreatic origin, but on no occasion did the increase approach the cutoff levels described for severe acute pancreatitis (>100 nmol/l). However, irAT levels in serum and urine were increased (>50 micro g/l) in 54% of the cases. CONCLUSION: Contrary to what is found for irAT, patients with acute abdominal pain of non-pancreatic origin rarely have markedly increased levels of irCAPAP in serum and urine. PMID- 12748425 TI - Oxygen radicals promote ICAM-1 expression and microcirculatory disturbances in experimental acute pancreatitis. AB - BACKGROUND/AIMS: The course of pancreatitis is paralleled by a drastic reduction in organ perfusion and increased ICAM-1-mediated leukocyte-endothelial interaction. We aimed to evaluate the effect of oxygen radicals on ICAM-1 expression and the microcirculation in severe acute pancreatitis using the oxygen radical scavenger dimethylsulfoxide (DMSO). MATERIALS AND METHODS: Severe pancreatitis was induced in rats (n = 32) who were randomly assigned to one of two groups: either 4 ml/kg 50% DMSO/saline (v/v) started 3 h after induction of pancreatitis or 4 ml/kg saline (control). Microcirculation was evaluated by intermittent intravital microscopy. Serum amylase and lipase, histomorphometric changes, immunohistochemistry for ICAM-1 expression and 24-hour survival were investigated. RESULTS: Leukocyte adherence was significantly reduced (4.4 +/- 0.47 vs. 5.58 +/- 0.69 sticker/100 micro m, p < 0.05), and mean capillary (0.96 +/- 0.06 vs. 0.45 +/- 0.13 mm/s; p < 0.01) and venous erythrocyte velocity (1.16 +/- 0.12 vs. 0.58 +/- 0.16 mm/s, p < 0.01) were significantly increased by DMSO treatment. Microcirculatory disturbances were paralleled by an increase in endothelial ICAM-1 expression, whereas DMSO reduced ICAM-1 expression. CONCLUSION: DMSO improves pancreatic microcirculation and reduces ICAM-1 expression and subsequent leukocyte adhesion, suggesting an important role of oxygen free radicals in the pathway of endothelial ICAM-1 expression and microcirculatory disturbances in acute pancreatitis. PMID- 12748426 TI - Effects of octreotide pretreatment in experimental acute pancreatitis. AB - BACKGROUND: Severity of systemic lesions and mortality of experimental acute pancreatitis (AP) are reduced after pancreatic enzyme content reduction induced by cerulein administration. Octreotide has been used both prophylatically and therapeutically in AP. The possible effects of octreotide on pancreatic enzyme content and its influence on pulmonary lesions of experimental AP were assessed in this study. METHODS: Wistar male rats were divided in two branches: BRANCH I - Animals divided into three groups: Group Sa (n = 10) intravenous saline infusion; Group Ce (n = 10) intravenous cerulein infusion, (0.133 micro g/kg(-1).h(-1)) and Group Oc (n = 10) SC octreotide (10 micro g/kg(-1)). Trypsin, elastase and amylase pancreatic contents as well as serum amylase were determined thereafter in all three groups; BRANCH II - Rats treated as in branch I, were submitted to sodium taurocholate AP (Groups Sa+AP, Ce+AP and Oc+AP). Two hours thereafter amylase and TAP assays were performed in serum, ascites and pancreatic tissue in eight animals of each group. Pulmonary histology was studied by morphometry 24 h after AP in the remaining animals. RESULTS: Increased serum amylase and pancreatic enzyme contents were observed in octreotide-treated animals when compared to animals receiving saline or cerulein. After AP increases of serum and ascitic fluid amylase and of pancreatic TAP were observed in octreotide pre treated animals when compared to saline and cerulein groups. Pulmonary interstitial and alveolar edema after AP was significantly increased in rats receiving octreotide as compared to the cerulein group. CONCLUSION: Octreotide administration acutely increases the enzymatic content of the pancreas and thus may have a potential deleterious influence in the evolution of AP. PMID- 12748428 TI - Perspectives of evidence-based surgery. AB - The assessment of the optimal treatment option based on best current knowledge is called evidence-based medicine (EBM). Considering the cost explosion in public health systems, EBM should also incorporate proper utilization of the restricted economical resources and should enforce quality assurance in medicine. It is imperative that surgeons realize that randomized controlled trials are applicable to the operative specialties in a large scale, and are necessary to provide evidence-based surgery. So far, only 3.4% of all publications in the leading surgical journals are randomized controlled trials. Furthermore, only 44.1% of the published surgical randomized controlled studies compared different surgical procedures, whereas 55.9% of the articles compared medical therapies in surgical patients. Evidence-based surgical therapy is essential for further development of a high- quality surgical standard, which will also provide quality assurance in future surgical care. This article presents the definition of EBM and discusses specific problems involved in the introduction of its principles into the surgical discipline. PMID- 12748427 TI - Systematic isolation of genes differentially expressed in normal and cancerous tissue of the pancreas. AB - BACKGROUND: There is increasing knowledge about the genetic basis of pancreatic cancer (PaCa). Tumor suppressor genes (TSGs; e.g. p53 and DPC4) and oncogenes (e.g. K-ras) have been shown to be involved in the development of PaCa. However, the extent of chromosomal changes (gains and losses) implicates that many more genes may be involved in the multistep progression of PaCa. Identification of these genes is essential for understanding the molecular events in the development of PaCa. METHODS: We assembled public and proprietary libraries of more than 4 million expressed sequence tags using newly developed software tools. RESULTS: We identified a total of 249 genes with specific expression patterns in normal and cancerous tissue of the pancreas. Of these, 27 genes were found to be preferentially expressed in normal tissue of the pancreas, while 222 genes showed significant upregulation of expression in PaCa. Of the 249 genes, 232 (93.2%) were found to represent known human genes or putative human homologues of genes characterized previously in other species, while 17 (6.8%) represent putative new genes. CONCLUSION: These genes may represent a valuable source to identify novel TSGs and oncogenes involved in the carcinogenesis of PaCa. PMID- 12748430 TI - Giant sclerosing mesenteritis. PMID- 12748429 TI - Minimally invasive retroperitoneal pancreatic necrosectomy. AB - INTRODUCTION: Open surgery for pancreatic necrosis is associated with considerable morbidity and mortality. We report the results of a recently developed minimally invasive technique that we adopted in 1998. METHODS: A descriptive explanation of the approach is given together with the results of a retrospective analysis of patients who underwent a minimally invasive retroperitoneal pancreatic necrosectomy (MIRP) between August 1998 and April 2002. PATIENTS: There were 24 patients with a median (range) age of 61 (29-75) years. The initial median (range) APACHE II score was 8 (2-21). All patients had infected pancreatic necrosis with at least 50% pancreatic necrosis. In three patients it was not possible to complete the first MIRP because of technical reasons. RESULTS: A total of 88 procedures were performed with a median (range) of 4 (0-8) per patient. Twenty-one (88%) patients developed 36 complications during the course of their illness. Five patients required an additional open procedure: 2 for subsequent distant collections, 2 for bleeding and 1 for persisting sepsis and a distant abscess. Six (25%) patients who had MIRP died. The median (range) post-operative hospital stay was 51 (5-200) days. CONCLUSIONS: MIRP is a new technique that has shown promising results, and could be preferable to open pancreatic necrosectomy in selected patients. However, unresolved issues remain to be overcome and the exact role of MIRP in the management of pancreatic necrosis has yet to be defined. PMID- 12748431 TI - A retrospective study comparing the different surgical procedures for the treatment of hydatid disease of the liver. AB - BACKGROUND/AIMS: The purpose of this study was to compare the security and effectiveness of the different surgical procedures used for treatment of the liver infestations with larvae of the dog tapeworm (Echinoocccus granulosus). METHODS: 55 patients with echinococcal cysts in the liver were treated surgically at our department between 1987 and 1999. The patients were grouped according to the mode of surgical technique and comprised 16 anatomical liver resections, 26 pericystectomies and 13 endocystectomies. The results of the different procedures have been analyzed statistically. RESULTS: The decision which procedure was to be performed was based on the severity of the disease. Complications requiring relaparotomy occurred in 5% of all cases, not showing any predisposition to a certain technique. Blood loss and duration of hospitalization was lowest in the endocystectomy group; however, the differences were not statistically significant. At a median follow-up period of 84 months, recurrences were noted in 2 cases (4%), 1 after pericystectomy and 1 after resection. CONCLUSION: There was no substantial evidence that the more radical procedure carries a lower risk of recurrence, and there was convincing evidence that endocystectomy is the most careful procedure in regard to blood loss and duration of hospitalization. PMID- 12748432 TI - Randomized clinical trial of postoperative subfascial infusion with bupivacaine following ambulatory open mesh repair of inguinal hernia. AB - BACKGROUND: Wound pain remains the commonest problem after ambulatory open repair of inguinal hernia. Postoperative subfascial infusion of the wound with bupivacaine extends local analgesia at home and may achieve superior analgesia compared with oral analgesics alone. The objective of the present trial was to evaluate the efficacy of postoperative subfascial infusion of the wound with 0.5% bupicavaine at 2 ml per hour for 48 h after operation. METHODS: Forty-four patients who underwent ambulatory open tension-free mesh hernioplasties were randomized to two arms of treatment. The pump group had an infusion pump containing 100 ml 0.5% bupivacaine being placed between the external oblique aponeurosis and the Prolene mesh, whereas the nonpump group was treated with oral analgesics alone. Assuming that an observed difference of 2.0 existed between the mean pain scores of the two groups, the estimated sample size would be at least 20 patients in each group. RESULTS: Postoperative pain scores at rest and on coughing were significantly lower in the pump group than in the nonpump group on days 0 and 1 after surgery (p < 0.01). Before being discharged, none of the pump group patients requested analgesics, but 6 patients of the nonpump group required analgesic supplement (p = 0.025). Ten patients (50%) of the pump group experienced no pain during the period of bupivacaine infusion. Recovery variables, including time taken to resume ambulation and micturition, were comparable between the two groups. The pump and nonpump group patients returned to their normal activities after a median of 3 and 4 days, respectively (p = 0.217). The postoperative morbidity rates of the two groups were similar. CONCLUSION: Postoperative subfascial infusion of the wound with 0.5% bupivacaine achieved superior analgesia compared with oral analgesics alone. Portable infusion pump is a safe technique to continue local analgesia at home after ambulatory open repair of inguinal hernia. The drawbacks of the ON-Q Pain Management System included its high cost and frequent seepage of blood-stained anesthetic fluid into the wound dressing. PMID- 12748433 TI - Offering preconceptional cystic fibrosis carrier couple screening in the absence of established preconceptional care services. AB - OBJECTIVE: To investigate the feasibility and acceptability of different modes of offering preconceptional carrier screening for cystic fibrosis (CF) in the absence of established preconceptional care services. METHODS: Individuals aged 20-35 years were invited by mail, either by the Municipal Health Services (MHS) or by their own general practitioner (GP) to participate in a screening program with their partner. Pretest education was provided either during a group educational session or during a GP consultation. The reasons given by participants and nonrespondents for (not) responding to the invitation for screening, sociodemographic characteristics, and their attitudes were assessed by means of questionnaires. RESULTS: Of 38,114 individuals who received a first invitation, approximately 20% had a partner with whom they were planning to have children. The response rate, as measured by attendance at either a group educational session or a GP consultation, was not affected by whether the letter was sent by the MHS or the person's GP. However, the response rate was about 2.5 times higher when the letter invited people to make an appointment with their GP for a consultation regarding CF carrier screening than when it invited them to attend an educational group session. A total of 559 couples (96%) consented to have the test after education. Repetition of the invitation increased the response. The main reason given by couples for not responding was "lack of time to attend" or "forgot about it" (48%). Another reason given was that they did not want to know their test results (28%). Eighty-nine percent of participants and 69% of nonrespondents believed that screening should be offered routinely to couples planning to have children. The GPs consulted (n = 18) reported no negative experiences, but due to the extra workload, 11 of them would not consider it to be part of their task. CONCLUSIONS: Among couples planning to have children, there is generally a positive attitude towards routinely offering population-based CF carrier screening. Preconceptional CF carrier screening appeared feasible, both in terms of practical achievements and target group accessibility. Participation varied according to the pretest education setting, with the primary care setting producing the highest rate of attendance. PMID- 12748434 TI - The simultaneous presence of alpha- and beta-thalassaemia alleles: a pitfall of thalassaemia screening. AB - OBJECTIVE: To compare the efficacy of routine haematological tests and molecular analysis in the diagnosis of double heterozygous alpha- and beta-thalassaemia. METHODS: Screening was carried out in extended family members from 125 families registered in the National Thalassaemia Registry, known to have both alpha- and beta-thalassaemia carriers. RESULTS: Eighty-three individuals from 59 families were identified to be double heterozygous for alpha- and beta-thalassaemia only upon molecular analyses. Among 40 married individuals, 1 was at 25% risk for having beta-thalassaemia major children and 6 for having Bart's hydrops pregnancies. CONCLUSION: Molecular analysis must be used for the accurate diagnosis of double heterozygous alpha- and beta-thalassaemia for proper risk ascertainment, especially in regions with a high prevalence of both types of thalassaemia. PMID- 12748435 TI - Public knowledge regarding the role of genetic susceptibility to environmentally induced health conditions. AB - OBJECTIVE: Diseases thought to be caused by exposure to environmental factors are also influenced by genetic susceptibility. It is not clear to what extent the public recognizes the role of genetics in causing these diseases. METHODS: We asked 2,353 people in a national survey to indicate their level of agreement with statements about the genetic contribution to four health conditions typically considered to be environmentally induced. RESULTS: 206 (9%) respondents believed that genetic susceptibility contributes to all four health conditions, while 751 (32%) believed that genetics plays no role in causing any of the conditions. Respondents were more likely to believe that genetics contributes to adverse drug reactions and smoking-related illnesses than to infectious diseases and diseases resulting from exposure to environmental agents. CONCLUSIONS: This study suggests that the public views genetic susceptibility as playing only a limited role in human disease induced by environmental factors. Increasing awareness of the role of genetic factors in these diseases will be necessary for translating gene discovery into effective personal and public health actions. PMID- 12748436 TI - Attitudes towards genetic screening for predisposition to colon cancer among cancer patients, their relatives and members of the community. Results of focus group interviews. AB - OBJECTIVE: To compare knowledge of and interest in genetic testing for hereditary colon cancer syndromes. METHODS: Colorectal cancer patients, first-degree relatives of colon cancer patients and controls were recruited from a familial cancer registry. Focus groups explored attitudes about genetic testing. RESULTS: All three groups conveyed interest in testing, but lacked knowledge about testing and its implications. After receiving information regarding the potential benefits and costs of testing (including insurance and employment issues) all three groups were disinclined to be tested. The reasons varied among risk groups. CONCLUSIONS: When informed about the costs and implications of testing, individuals may be reluctant to undergo genetic testing, regardless of baseline risk. Barriers to testing will vary depending on the perceived risk of carrying a mutation. PMID- 12748437 TI - Genebanks: a comparison of eight proposed international genetic databases. AB - OBJECTIVE: To identify and compare population-based genetic databases, or "genebanks", that have been proposed in eight international locations between 1998 and 2002. A genebank can be defined as a stored collection of genetic samples in the form of blood or tissue, that can be linked with medical and genealogical or lifestyle information from a specific population, gathered using a process of generalized consent. METHODS: Genebanks were identified by searching Medline and internet search engines with key words such as "genetic database" and "biobank" and by reviewing literature on previously identified databases such as the deCode project. Collection of genebank characteristics was by an electronic and literature search, augmented by correspondence with informed individuals. The proposed genebanks are located in Iceland, the United Kingdom, Estonia, Latvia, Sweden, Singapore, the Kingdom of Tonga, and Quebec, Canada. Comparisons of the genebanks were based on the following criteria: genebank location and description of purpose, role of government, commercial involvement, consent and confidentiality procedures, opposition to the genebank, and current progress. RESULTS: All of the groups proposing the genebanks plan to search for susceptibility genes for complex diseases while attempting to improve public health and medical care in the region and, in some cases, stimulating the local economy through expansion of the biotechnology sector. While all of the identified plans share these purposes, they differ in many aspects, including funding, subject participation, and organization. The balance of government and commercial involvement in the development of each project varies. Genetic samples and health information will be collected from participants and coded in all of the genebanks, but consent procedures range from presumed consent of the entire eligible population to recruitment of volunteers with informed consent. Issues regarding confidentiality and consent have resulted in opposition to some of the more publicized projects. None of the proposed databases are currently operational and at least one project was terminated due to opposition. CONCLUSIONS: Ambitious genebank projects have been proposed in numerous countries and provinces. The characteristics of the projects vary, but all intend to map genes for common diseases and hope to improve the health of the populations involved. The impact of these projects on understanding genetic susceptibility to disease will be increasingly apparent if the projects become operational. The ethical, legal, and social implications of the projects should be carefully considered during their development. PMID- 12748438 TI - Where there's a web, there's a way: commercial genetic testing and the Internet. AB - The Internet has become a "global marketplace", enabling consumers to purchase health care products and services, including genetic testing, through a variety of national and international sources. A web search for commercial (for-profit) genetic testing companies found 12 with a web presence that were offering adult genetic susceptibility testing, of which 3 offered direct-to-consumer access. In this paper, Canada--with its educated population and universal health care system -will serve as a case study for illustrating the social, ethical and policy issues (e.g., information privacy, just access to health care, product safety, and access to unbiased health information) arising with Internet-based access to commercial genetic testing. Health professionals, policy makers and consumers in all developed nations will be faced with complex technical, social and ethical issues, but without further discussion it will not be possible to determine how best to manage and maximise the benefits of this increased accessibility and choice, while minimising the associated personal and social costs. PMID- 12748439 TI - Frequency of consanguineous marriages in patients with hereditary blood disorders in southern Turkey. PMID- 12748440 TI - [Salmonella non-typhoid infection: new epidemiological findings] AB - Food-borne infections are the most serious food safety problem in the world. In fact, they are responsible of millions of illnesses and thousand of deaths. Non typhoid Salmonella infection is frequent world-wide and, although mild and self limiting illness in normal subjects, it may cause a severe disease in patients with an immune-deficiency. Changes in the agents and in the vehicles of transmission and a higher number of patients with immune-depression have determined a world-spread of non-typhoid Salmonella infection in the last decades. The increased frequency of international travels and food commerce have been associated with outbreaks of unusual serotype of Salmonella. Moreover, drug resistant Salmonella are emerged recently, as Ampicillin and Doxiciclin-resistant S. enteritidis or DT-104 multidrug-resistant S. typhimurium. The outbreak of Salmonella disease is also linked to diffusion of HIV infection and of other immunodeficiencies. The lack of controls in food industry, the frequent contamination of mass-distributed food products and the decreased opportunities to transmit for instruction on food safety, both in school and inside the family, are the causes of large-scale outbreaks PMID- 12748441 TI - Value of the test for serum HCV-RNA in clinical practice. AB - The test to identify genomic RNA is the only tool currently available to directly evidence the presence of the hepatitis C virus (HCV) in infected subjects. In this review we examined the most commonly used qualitative and quantitative methods to detect HCV-RNA in serum or plasma, focusing particular attention on commercially available methods. Furthermore, we report the clinical conditions in which the viremia test is useful or even mandatory for the diagnosis of HCV infection, and comment on the usefulness of the test for monitoring patients in the course of antiviral therapy. PMID- 12748443 TI - [Bacterial pneumonia in HIV-infected patients] AB - Patients and methods: This is a retrospective study performed on HIV-positive patients discharged from our Institution from January 1993 through December 1998 with a diagnosis of bacterial pneumonia. Cases of TB or atypical micobacterial infection were excluded from this analysis. Causative organisms were identified, when possible, by taking into account positive cultures from diverse sources (blood, sputum, pleural fluid and others). Results: In the 6-yr period we considered, 120 patients were identified. Among them, we were able to obtain clinical and imaging data on 98 cases. Focal infiltrates on chest X-ray studies were present in 87.7% of cases, 24.5% had a pleural effusion, 9.2% nodular lesions, 4.1% cavitary images and in 2 patients only hilar lymphadenopaties were noted. Causative agents resulted to be S. aureus (14 cases); P. aeruginosa (11); S. pneumoniae (6); R. equi (4); Enterobacter spp. and K. pneumoniae (3 cases each); E. coli, Peptostreptococcus spp and Enterococcus spp. (1 case each). No causative organism was isolated in 54 patients (55.1%) and the diagnosis was based on clinical and therapeutical grounds. Around half of cases (46.9%) responded well to therapy, while 11 (11.2%) died because of the lung infection. In 3 cases other opportunistic infections were the cause of death and 22 cases of relapse were recorded as well. Five patients resulted lost to follow-up. Conclusions: This retrospective study demonstrated a high prevalence of S. aureus lung infections along with the presence of otherwise rare causative organisms such as R. equi. Radiologic appearance of lung lesions did not differ substantially from the one of HIV negative patients. A relatively good response to antibiotic therapy was also noted. PMID- 12748442 TI - [Central venous catheter-related sepsis: one year experience in a large university hospital] AB - Five hundred two central venous catheters (CVC) inserted in 366 patients were evaluated prospectively over a one-year period to determine the frequency and risk factors associated with CVC-related sepsis. A CVC-related infection was suspected in 190 cases (190/502, 38%). A catheter-related sepsis was established in 50 patients (10%). The infection rate was 0.8 cases of sepsis per 100 catheterdays. Staphylococcus epidermidis, Staphylococcus aureus, and Candida spp. were the most frequently isolated etiological agents of sepsis. On univariate analysis, six variables affecting the rate of catheter-related sepsis were identified: neutropenia > 8 days (p < 0.01); AIDS (p < 0.001); haematological malignancies (p < 0.001); total parenteral nutrition (p = 0.001); duration of site use (p = 0.04); high APACHE II score (p = 0.04). The logistic regression analysis revealed that AIDS and haematological malignancies were independent risk factors of catheter-related sepsis. In conclusion, although the incidence of established catheter infection is much lower than the incidence of suspected infection, in most cases it is wise to change the catheter with the guidewire technique and wait for culture of the tip, rather than to remove the catheter immediately. PMID- 12748444 TI - [Sepsis: our series] AB - Sepsis understood as persistent bacteremia with considerable clinical symptomatology that can develop into septic shock, retains its clinical, diagnostic and therapcutic meaning. Its most recent definition as a systemic inflammatory secondary reaction to a verified infection may help us to understand the reactive events of the host against infections and prevent septic shock. We report clinical cases of sepsis with positive haemoculture observed in the Caltagirone (CT) Hospital in the last 15 years: 186 patients (117 males and 69 females), with particular focus on those over 61 years old and on patients admitted to the ICU. The clinical isolates and groups of the inpatients are listed. In particular, two cases of toxic shock syndrome and six cases of bacterial endocarditis are described. Mortality was about 30% for septic shock despite rational antibiotic therapy, support therapy and hospital admission to the intensive care unit PMID- 12748445 TI - [Correlation between CD4 T-cell counts and HIV-1 RNA plasma levels in HIV-1 patients receiving highly active antiretroviral therapy (HAART)] AB - The prognostic value of plasma HIV-1 RNA baseline levels in patients who are going to receive HAART has been recently questioned. In the present study the authors correlated the baseline counts of viremy and CD4 with the viral suppression induced by HAART in an ongoing cohort of HIV-1 positive patients. Data resulting from the study suggest that the HAART effect on CD4 T-cells depends both on the immunological status before starting therapy and on the degree of viral suppression. After briefly discussing about the possible causes of disconnection between CD4 T-cells count and plasma HIV-1 RNA levels, authors conclude that the viral suppression is the desired goal of antiretroviral treatment and that the maximum effect of HAART can be achieved by carefully clinically evaluating patients and individuating the best therapy. PMID- 12748446 TI - [Microbiological survey in an Intensive Care Unit] AB - An epidemiological survey on 417 sample from ICU patients, was carried out from January '97 to December 98 to verify the epidemiology of bacteria responsible for nosocomial infections and to monitor the susceptibility patterns to antimicrobial drugs during this period. Microbiological cultures were positive in 47.7% of the samples. P. aeruginosa was the most frequent pathogen, followed by Staphylococcus aureus, Candida spp, coagulase-negative staphylococci, E.coli, with a clear prevalence of Gram-negative over Gram-positive isolates. The evaluation of the antibiotic susceptibility profile of this bacterial pathogens suggests the importance of a costant epidemiological surveillance in ICU. PMID- 12748447 TI - [Eosinophilia and Strongyloides stercoralis infestation: case report] AB - Authors report about a case of Strongyloides stercoralis infestation. The patient, a 65-year old man, presented with a clinical history of eosinophilia but without symptomatology. Several stool specimen showed the presence of rhabditiform larvae of S. stercoralis and after therapy the patient had a normalization of the white blood ceIls count. The authors wish to point out the importance of the diagnosis because a change in immune status may convert a previously asymptomatic infection to hyperinfection. PMID- 12748448 TI - [A case of sepsis caused by BCG (Bacillus Calmette Guerin) after its bladder instillation in a 71-year-old patient affected by bladder carcinoma] AB - Sometimes the Calmette-Guerin bacillus can be used with success as local immunotherapy for superficial bladder carcinoma. BCG is a living attenuated strain of Mycobacterium bovis and therefore its biadder instillation may give rise, in some particular cases, to serious systemic effects, caused either by hypersensitivity reaction or by systemic dissemination in case with low efficiency of cellular immunity. We describe a case of sepsis in a 71-year-old patient after bladder instillation of BCG with a rare complication of granulomatous hepatitis. The etiological diagnosis of granulomatous hepatitis with PCR on liver biopsy is very important for the specific therapy before the patient can undergo a steroid regime. PMID- 12748449 TI - [Plaut-Vincent's angina] AB - The clinical appearance of fuso-spirillar infection in patients with a serious impairment of the immunitary system was the starting-point for a historical study. At the end of nineteenth century two european researchers clarified the cause of many infections. Association of a spindle-shaped bacillus with a spirillar one was shown to cause several pathologies: necrotic ulcerous stomatitis, pseudomembranous angina, hepatic abscess, otitis, pneumonia, etc. The identification of such bacteria contributed to improving further differential diagnosis with diphteria PMID- 12748450 TI - A new image. PMID- 12748451 TI - Herbs commonly used by women: an evidence-based review. AB - OBJECTIVE: To review the evidence of herbs commonly used by women. DATA SOURCES: Articles were located by searching Medline, Cochrane Database of Systemic Reviews, and the Combined Health Information Database and by hand searching the reference lists of recent systematic reviews. The databases were searched in January 2000 and October 2000 by using the Latin and common name of each herb. METHODS OF STUDY SELECTION: Preference was given to randomized, placebo controlled trials. When available, English language studies were reviewed. If not, data are presented from review articles that summarize the foreign study. RESULTS: Many women use herbal therapies. In the United States, herbs are considered dietary supplements. The Food and Drug Administration (FDA) cannot remove them from the market unless they are proven unsafe. The herb industry plans to improve monitoring. Many prospective randomized controlled trials are being funded. Gingko biloba seems to slow the progression of dementia but increases the risk of bleeding. St John's Wort is efficacious for treating mild to moderate depression but has many drug interactions. Ginseng seems to improve well being in perimenopausal women, but it is often impure and has side effects and drug interactions. Garlic slightly lowers blood pressure and lipids. Echinacea slightly decreases the duration of colds but does not prevent them. Valerian is beneficial for insomnia, but there is no long-term safety data. Black cohosh may help the symptoms of perimenopause, and chasteberry may improve premenstrual syndrome. More study is needed on both herbs. CONCLUSION: Some herbs are medically useful, but the American public would benefit from increased regulation. Manufacturers should be able to ensure that herbs contain pure ingredients. Side effects and drug interactions should be listed. Well-designed studies are being conducted. The results will be helpful to physicians and patients when the clinical evidence becomes available. PMID- 12748452 TI - An integrative medicine approach to premenstrual syndrome. AB - Complementary and alternative medicine (CAM) approaches are widely used by women with premenstrual syndrome (PMS) and premenstrual dysphoric disorder (PMDD). This article provides a comprehensive review of the medical literature on clinical applications of CAM for these conditions. The information was collected via a Medline review dating back to 1966 and subsequent selected review of bibliographies from these articles for non-Medline referenced but relevant clinical studies. For many of the therapies discussed, there is a lack of conclusive evidence either confirming or refuting efficacy. For other therapies, including certain herbal and nutritional approaches, the use of exercise, and the use of mind-body approaches, there is substantial evidence of efficacy. This review will be relevant to the practicing clinician seeking to become aware of and to understand the relevance of the complementary/alternative therapies being used by his/her patients for PMS and PMDD. PMID- 12748453 TI - Phytoestrogens and breast cancer prevention: what is the evidence? AB - The use of phytoestrogens as a natural source of estrogen has been rapidly accepted by consumers for various remedial or preventive purposes, including breast cancer prevention. Phytoestrogens exhibit mixed weak estrogen agonist/antagonist properties and antioxidative activity. They may play a significant inhibitory role during the initiation and promotional phases of cancer development. Experimental and physiologic studies provide much of the evidence on the inverse association between breast cancer risk and phytoestrogen intake. Research in humans has been limited to observational (case-control) epidemiologic studies and is far from conclusive. A critical evaluation through controlled trials of phytoestrogens' breast cancer-protective role needs to be performed before they are adopted as chemopreventive agents. PMID- 12748456 TI - Progestins and breast cancer. AB - The relationship between the use of menopausal hormone therapy (ERT, unopposed estrogen therapy; HRT, combined estrogen and progestin therapy) and the development of breast cancer remains controversial. Mechanistic studies examining progestins in human breast cancer cell lines have demonstrated a biphasic cellular response to progesterone; initial exposure to hormone results in a proliferative burst with sustained exposure resulting in growth inhibition. To date, there is no definitive evidence that progestins act in the pathogenesis of breast cancer. Epidemiologic studies have produced inconsistent results, and data from randomized, placebo-controlled trials are limited. Although recent results from the continuous combined therapy arm of the Women's Health Initiative trial showed a small increase in the risk of invasive breast cancer in women on therapy for 5 years or more, a clear consensus regarding the relationship between HRT and breast cancer risk cannot yet be drawn from existing data. Studies have consistently documented that HRT use is associated with improved mortality and survival rates for women with breast cancer. Large-scale, randomized studies on different progestin regimens are needed to critically assess the effect of progestin on breast cancer. PMID- 12748454 TI - Early identification of mild traumatic brain injury in female victims of domestic violence. AB - BACKGROUND: Domestic violence and traumatic brain injuries (TBIs) are major societal problems with public health implications. The inter-relationship between this specific form of violence and TBI has not been explored. However, a clinically relevant relationship is suggested by recent outcome data showing that TBI survivors who are victims of assault are more prone to a suboptimal outcome. METHODS: This observational study screened women who presented to 3 metropolitan emergency departments for health issues associated with domestic violence for a possible secondary mild TBI. RESULTS: Over a 7- to 9-month survey period, a total of 169 women with health issues directly attributable to domestic violence and who were referred to sexual assault-domestic violence health staff were identified. Thirty percent (n = 51) agreed to participate in this survey. A positive loss of consciousness on at least one occasion was reported in 30% of the respondents, and 67% of women reported residual problems that were potentially head-injury related. CONCLUSION: These data argue for greater vigilance in screening domestic violence victims for mild TBI because earlier identification and treatment of any postconcussive syndrome symptoms seems to diminish their adverse impact on outcome. PMID- 12748458 TI - Innervation of the levator ani muscles in the female squirrel monkey. AB - OBJECTIVE: The purpose of this study was to characterize the innervation of the levator ani muscles in the female squirrel monkey and to investigate its usefulness as an animal model of pelvic organ prolapse. STUDY DESIGN: Eleven nulliparous female squirrel monkeys with no pelvic organ prolapse were used in this study. Detailed pelvic dissections were conducted (n = 3), and the Koelle stain for acetylcholinesterase was used to identify the motor endplate zone in the levator ani muscles (n = 2). Unilateral levator ani (n = 4) and pudendal (n = 2) neurectomies were performed; changes in levator ani muscle mass and myocyte diameter were examined 14 days after neurectomy. Nerve biopsy specimens from each animal were processed for microscopy. RESULTS: The levator ani nerve originated from the S2 spinal root and entered the pelvic cavity adjacent to the pelvic nerve between the flexor caudalis brevis and iliocaudalis muscles. The levator ani nerve then projected caudally and bifurcated to penetrate the iliocaudalis and pubocaudalis. A single motor endplate zone in each muscle correlated with the point of levator ani nerve penetration. The pudendal nerve originated from the S1 S2 spinal roots to innervate the urethral and anal sphincters, clitoris, and perineum, but not the iliocaudalis or pubocaudalis. Significant atrophy and myocyte shrinkage occurred in the iliocaudalis and pubocaudalis ipsilateral to the levator ani nerve transection (P <.05). Pudendal neurectomy produced no levator ani muscle changes. CONCLUSION: Intrapelvic skeletal muscles in the female squirrel monkey are similar to humans and have distinct innervation with no contribution from the pudendal nerve. The squirrel monkey is likely to be a useful model of pelvic organ prolapse and warrants further study. PMID- 12748457 TI - Contrasting effects of two hormone replacement therapies on the cardiovascular and mammary gland outcomes in surgically postmenopausal monkeys. AB - OBJECTIVE: The purpose of this study was to compare the effects of two hormone replacement therapies on the intermediate end points of coronary heart disease and mammary gland hyperplasia in postmenopausal monkeys. STUDY DESIGN: Surgically postmenopausal cynomolgus monkeys were fed an atherogenic diet for 12 months while receiving no treatment (control, n = 19), conjugated equine estrogens plus continuous medroxyprogesterone acetate (n = 19), or ethinyl estradiol plus norethindrone acetate (n = 21) at doses that were scaled from those doses taken by women. RESULTS: Quantitative coronary angiography revealed that the arteries of the control group and the conjugated equine estrogens plus continuous medroxyprogesterone acetate-treated animals constricted in response to acetylcholine (-5.4% +/- 1.4% and -6.2% +/- 1.5%, respectively), whereas those arteries in the animals in the ethinyl estradiol plus norethindrone acetate group did not (P =.002). The incidence of dobutamine-induced ST-segment depression in the ethinyl estradiol plus norethindrone acetate group (10.5%) was significantly less than in the control group (68.8%, P =.001) or the conjugated equine estrogens plus continuous medroxyprogesterone acetate group (50%, P =.01). Conjugated equine estrogens plus continuous medroxyprogesterone acetate, but not ethinyl estradiol plus norethindrone acetate, induced diffuse epithelial tissue proliferation in the mammary glands (P =.0006). CONCLUSION: Ethinyl estradiol plus norethindrone acetate protected against atherosclerosis-induced endothelium mediated vasoconstriction of coronary arteries and heart rate-induced myocardial ischemia and did not induce epithelial tissue proliferation (tissue density) in the mammary gland. PMID- 12748459 TI - Predominance of serotype V and frequency of erythromycin resistance in Streptococcus agalactiae in Ohio. AB - OBJECTIVE: The purpose of this study was to test the serotype distribution and antibiotic sensitivity patterns of group B Streptococcus from three Ohio regions in comparison to other areas of the United States. STUDY DESIGN: Three hundred forty-nine group B Streptococcus isolates from three Ohio hospitals were serotyped specifically, and disk diffusion was used to determine antibiotic susceptibility. RESULTS: Serotype V was isolated most frequently (27%); major types Ia, Ib, II, and III had frequencies of 18%, 9%, 11%, and 17%, respectively. Erythromycin and clindamycin resistance was 12% (n = 200 isolates) and 8% (n = 49 isolates), respectively. CONCLUSION: The prevalence shift of resistant serotype V group B streptococci in Ohio reflects regional increases and suggests that alternative therapies for patients who are allergic to penicillin may be inadequate. PMID- 12748460 TI - Postmolar surveillance at a trophoblastic disease center that serves indigent women. AB - OBJECTIVE: The purpose of this study was to determine compliance with postmolar pregnancy surveillance in our indigent population. STUDY DESIGN: Data for all women who were diagnosed with molar pregnancy from January 1996 through December 2000 were entered prospectively into a database. After remission, postmolar pregnancy surveillance was continued for 6 months. Patients whose condition required chemotherapy for gestational trophoblastic tumor had 12 months of follow up. Medical records were reviewed. RESULTS: Molar pregnancies occurred in 121 women: 103 Hispanic women (85%), 12 African American women (10%), and 6 white women (5%). Eighty-two women (68%) achieved remission without chemotherapy; 23 women (19%) were lost to follow-up without achieving remission, and 16 women (13%) had gestational trophoblastic tumor. Fifty-six Hispanic women (54%) completed postmolar pregnancy surveillance, compared with two African American women (11%, P <.01). Hispanic patients who were fluent in Spanish only were more likely to complete follow-up than bilingual Hispanic patients (62% vs 41%, P <.01). CONCLUSION: Hispanic women who were fluent in Spanish only were most likely to complete the recommended postmolar human chorionic gonadotropin surveillance. PMID- 12748461 TI - The role of covering gowns in reducing rates of bacterial contamination of scrub suits. AB - OBJECTIVE: This study was undertaken to determine whether covering gowns reduce the rates of contamination of surgical scrubs. STUDY DESIGN: Seventy-five clinicians had pieces of fabric from clean scrubs attached to two areas of their scrub suits. Participants wore a covering garment when wearing scrub suits off of designated areas (n = 25), did not wear a covering garment (n = 25), or wore scrub suits outside the hospital (n = 25). Subsequently, the fabric was assessed with culture in enhanced broth media and blood agar. RESULTS: Although there was a trend toward lower rates of contamination in the group that did not wear a covering garment, the difference was not significant. At no point, and at neither site of fabric attachment, did those who wore a covering garment demonstrate any advantage in regard to levels or frequency of contamination. CONCLUSION: Wearing covering garments over scrub suits does not reduce rates of contamination. PMID- 12748462 TI - Dual fluorescence analysis of DNA apoptosis in sperm. AB - OBJECTIVE: The objective was to compare fluorochrome Hoechst 33342 (Ho342) with combined Ho342/propidium iodide (PI) stains for assessment of sperm quality. STUDY DESIGN: Washed donor sperm cells were incubated in either 0, 0.15, or 15 micromol/L camptothecin (CAM) or 0.37 or 3.7 mmol/L genistein (GEN) for 4 hours at 37 degrees C. The sperm cells were analyzed for cycle-independent apoptosis and necrosis by single- compared with dual-stained fluorescence microscopy to contrast the relative effectiveness of these two approaches. RESULTS: The single stain procedure did not detect viability differences (overall 76.1% +/- 2.2% live). In contrast, the dual-stain procedure identified a dose-dependent decrease in viability and increased necrozoospermia for CAM and GEN treatments. Apoptosis was 2-fold higher with topoisomerase inhibitor treatment. CONCLUSION: The two topoisomerase inhibitors were associated with increased apoptosis and dose dependent necrosis. The data suggested that the dual-stain combination Ho342/PI was more sensitive than the single Ho342 stain analysis and permitted quantifying the apoptosis and necrosis events in sperm. PMID- 12748463 TI - A randomized controlled trial of second- versus third-generation oral contraceptives in the treatment of acne vulgaris. AB - OBJECTIVE: This study was undertaken to compare the clinical efficacy of second- versus third-generation oral contraceptives in the treatment of acne. STUDY DESIGN: Thirty-four women with acne were randomly selected to receive an oral contraceptive containing 0.3 mg of ethinyl estradiol (EE)/0.15 mg of desogestrel or 0.3 mg of EE/0.15 mg of levonorgestrel for 9 months. Acne was scored by lesion counting by a single examiner, and serum was analyzed for sex hormone-binding globulin (SHBG), dehydroepiandrosterone sulfate (DHEAS), and free and total testosterone at baseline and 3-month intervals. RESULTS: At baseline, the two treatment groups did not differ in the mean age, body mass index, acne lesion counts, SHBG, DHEAS, or free and total testosterone. Mean acne lesion counts decreased significantly in both groups from baseline (P <.02). In subjects completing 9 months of therapy, acne decreased by 52.8% in the EE/levonorgestrel group (n = 9) and by 58.5% in the EE/desogestrel group (n = 7) (between groups: P not significant). Mean SHBG increased by 46.3 nmol/L in the EE/desogestrel group (P not significant), and 20.0 nmol/L in the EE/levonorgestrel group (P <.05). Decreases in free testosterone from baseline occurred in each group, but these differences did not reach statistical significance. CONCLUSION: Oral contraceptives containing EE/desogestrel and EE/levonorgestrel were both effective in treating acne. PMID- 12748464 TI - The effect of training on the provision of elective abortion: a survey of five residency programs. AB - OBJECTIVE: We investigated the correlation of residency abortion training and other variables with abortion provision. STUDY DESIGN: We surveyed obstetrician gynecologists who graduated from five residency programs. Subjects were asked about demographic characteristics, residency abortion training, and abortion provision. We used a logistic regression model that included all variables that were correlated significantly with abortion provision in univariate analyses. RESULTS: Of 161 physicians (61%) who responded, 131 physicians (83%) participated in abortion training. Seventy-six physicians (47%) provided abortions currently, and 95 physicians (58%) had provided abortions at some time since residency. Three variables were independently, positively correlated with abortion provision: the number of abortions that were performed during residency, the gestational age limit of abortion training, and urban practice. Negatively correlated with provision were membership in a restrictive practice and training outside the teaching hospital. CONCLUSION: Variables that were indicative of more extensive residency abortion training were associated positively with the current provision of abortion. PMID- 12748465 TI - Local anesthetic reduces the pain of colposcopic biopsies: a randomized trial. AB - OBJECTIVE: This study was undertaken to determine whether administration of local anesthetic reduces pain associated with colposcopically directed cervical biopsies. STUDY DESIGN: Fifty-six patients undergoing colposcopically directed cervical biopsies were randomly selected prospectively to receive a 1% lidocaine injection or no injection before the biopsy procedure. Pain was assessed by using a 10-cm visual analog scale at various points during the procedure. Data were analyzed with the Student t test. RESULTS: Injection of lidocaine resulted in a reduction in pain scores for cervical biopsies (4.004 to 1.166, P: <.001), endocervical curettage (5.757 to 3.900, P =.0018), and overall procedure scores (4.911 to 3.145, P =.0013). CONCLUSION: This pilot study demonstrates that injection of local anesthetic at the site of colposcopically direct cervical biopsies decreases the pain perceived by patients. PMID- 12748466 TI - A comparison of two models of gynecology service consultation to the emergency department in an academic medical center. AB - OBJECTIVE: This study was undertaken to determine differences in resource use and outcomes when emergency department (ED) physicians consult the gynecology service routinely versus selectively. STUDY DESIGN: In July 2000, an ED policy of "routine" gynecology consultation for pregnant women less than 20 weeks' gestation with pain and/or bleeding complaints changed to a policy of "selective" consultation. Resource use and outcomes were compared for 222 women who received care during the 9 months before the protocol change with 268 women who presented during the 9 months after the protocol change. RESULTS: With selective consultation, patients receiving gynecology evaluations decreased from 74% to 39%. Return visits to the ED increased from 9% to 21%. Ultrasound studies performed by ED physicians and radiologists increased, whereas studies by gynecologists decreased. Patients waited longer and received more unnecessary human chorionic gonadotropin studies. CONCLUSION: A policy of selective gynecology consultation, compared with routine gynecology consultation in the ED, increases diagnostic study resource use and patient length of stay. PMID- 12748467 TI - Extremes of body mass index do not affect first-trimester pregnancy outcome in patients with infertility. AB - OBJECTIVE: The study was undertaken to ascertain whether body mass index (BMI) affects first-trimester pregnancy outcome in patients with infertility. STUDY DESIGN: Records of 494 patients with a singleton gestation after treatment for infertility at a single academic center were retrospectively reviewed. Patients were classified with regard to BMI and treatment protocol. Outcomes were defined as ongoing pregnancy, spontaneous abortion, or ectopic pregnancy. RESULTS: The average rate of spontaneous abortion was 21.5% and of ongoing pregnancy 75.1%. This did not differ significantly in any of the BMI groups. When patients were further subdivided according to diagnosis or fertility treatment, the spontaneous abortion rate remained the same. CONCLUSION: The likelihood of a spontaneous abortion, ongoing pregnancy, or ectopic pregnancy in singleton gestations in the first trimester, after treatment for infertility, was not affected by BMI. PMID- 12748468 TI - Characteristics predictive of response to ovarian diathermy in women with polycystic ovarian syndrome. AB - OBJECTIVE AND STUDY DESIGN: Ovarian needle diathermy was used in the treatment of clomiphene-resistant polycystic ovarian syndrome (PCOS). Factors predictive of pregnancy in our study group included younger age, lower body weight, and surgery before 1998. RESULTS AND CONCLUSION: Insulin resistance and adhesions at surgery were negatively associated with pregnancy. Sixty-six percent of the study group cycled spontaneously after surgery and 50% became pregnant. PMID- 12748469 TI - Expression of cyclooxygenase-2 in cervical, endometrial, and ovarian malignancies. AB - OBJECTIVE: This study was undertaken to compare the presence of cyclooxygenase-2 (COX-2) levels between normal and malignant cervix, endometrium, and ovary. STUDY DESIGN: Semiquantitative immunofluorescent assays for COX-2 were performed on sections of cervical squamous cell carcinoma (n = 12), endometrial adenocarcinoma (n = 13), and ovarian serous adenocarcinoma (n = 9). Levels of immunofluorescence for each sample were objectively measured, categorized as high, moderate, or negative expression and compared with normal cervical (n = 14), endometrial (n = 15), and ovarian (n = 13) tissue with the Fisher exact test. RESULTS: Normal cervical tissue expressed COX-2 more frequently than cervical cancer (50% vs 23%), but the difference was not significant (P =.247). COX-2 was rarely present in normal endometrium (7%) and normal ovarian epithelium (0%) and was usually present in endometrial adenocarcinoma (69%, P <.001) and ovarian serous cystadenocarcinoma (89%, P <.001). CONCLUSION: In this cohort, COX-2 expression is significantly more common in endometrial adenocarcinoma and ovarian serous cystadenocarcinoma, but not in cervical squamous carcinoma, compared with normal tissue. PMID- 12748470 TI - Cancer patients' satisfaction with physicians: Princess Margaret Hospital Satisfaction with Doctor Questionnaire results. AB - OBJECTIVE: The study was performed to examine the satisfaction of a specific population of oncology patients with their physicians and to quantify its association with characteristics of their disease. STUDY DESIGN: A descriptive design was used in which patients attending a weekend cancer support retreat completed the Princess Margaret Hospital Satisfaction with Doctor Questionnaire (PMH-PSQ-MD). Demographic information was requested separately. RESULTS: Of 48 patients, 96% completed the PMH-PSQ-MD, and 67% mailed in the additional demographics. Average overall score was 3.08 (SD = 0.56, 4 being most satisfied). Patients reported a desire for more time with physicians and that their pain be better understood. Patients were most satisfied with the physician's honesty, thoroughness, and communication. Satisfaction scores did not correlate with intensity of treatment or time since diagnosis. Scores declined as financial burden of therapy increased, but the trend was not statistically significant. CONCLUSION: Patients attending this retreat were generally satisfied with their physicians, regardless of treatment intensity. Increasing financial burden may be associated with physician dissatisfaction. PMID- 12748471 TI - Loop excision for high-grade squamous intraepithelial lesion on cytology: correlation with colposcopic and histologic findings. AB - OBJECTIVE: Our purpose was to examine the correlation between colposcopic and histologic findings in patients who have undergone loop electrosurgical excision of the cervix (LEEP) for high-grade dysplasia on cytology without prior colposcopically directed biopsy. STUDY DESIGN: A retrospective review was performed of all patients who underwent LEEP for high-grade squamous intraepithelial lesion (HGSIL) on Papanicolaou (Pap) smear without a prior cervical biopsy over a 17-month period. We correlated the histologic result with colposcopic findings at the time of LEEP. RESULTS: Of 104 patients undergoing LEEP, 63 patients (61%) had cervical intraepithelial neoplasia (CIN) grade 2 or greater. Thirty-four (54%) of these 63 patients with histologically proved high grade dysplasia had a normal or low-grade colposcopic examination. There were 10 patients with CIN 1 to CIN 1-2 and 1 patient with microinvasive squamous cell carcinoma. CONCLUSION: Despite lack of correlation between colposcopic and histologic results, HGSIL on Pap smear is an appropriate indication for LEEP. PMID- 12748473 TI - A cost-effectiveness analysis of management strategies for cervical intraepithelial neoplasia grades 2 and 3. AB - OBJECTIVE: This study was performed to compare the cost-effectiveness of strategies for the management of cervical intraepithelial neoplasia, grades 2 and 3 (CIN 2 and CIN 3). STUDY DESIGN: A decision analysis compared six management strategies. The model was applied to hypothetic cohorts of 100,000 patients with CIN 2 and CIN 3. RESULTS: In both analyses, cryotherapy (CRYO) was the least expensive and least effective strategy; total vaginal hysterectomy was the most expensive and most effective strategy. For CIN 2, loop electrosurgical excision procedure (LEEP) was more effective than CRYO but at a cost of $31,437 per additional cure and $1.8 million per additional cancer prevented. For CIN 3, LEEP was more effective than CRYO, but at a cost of $17,592 per additional cure and $1.0 million per additional cancer prevented. CONCLUSION: CRYO is a cost effective strategy that is appropriate in resource-poor settings. LEEP is also cost-effective, but the improved efficacy compared with CRYO comes at a significant cost. Total vaginal hysterectomy is very effective but is economically unsound because of excessively high costs. PMID- 12748472 TI - A technique to evaluate the thickness and density of nonvascular smooth muscle in the suburethral fibromuscular layer. AB - OBJECTIVE: The suburethral fibromuscular layer (SUFML) contains nonvascular smooth muscle that may play an important role in the continence mechanism. This study was undertaken to develop a method to systematically quantify differences in the thickness and density of nonvascular smooth muscle within the SUFML and to demonstrate its use in assessing variation. STUDY DESIGN: Urethral and anterior vaginal wall sections from the urethrovesical junction of fresh cadavers were stained to define alpha actin in smooth muscle cells. The thickness of the SUFML was microscopically measured and the density of the nonvascular smooth muscle was determined with computer-aided analysis. RESULTS: The mean thickness of the SUFML was 3.04 mm (range 1.74-3.78 mm, SD +/-0.63 mm). The mean density of nonvascular smooth muscle within it was 17.94% (range 10.48%-42.84%, SD +/-9.4%). The mean age was 55 years (range 36-69 years, SD +/-10.75 years). CONCLUSION: The thickness and density of the nonvascular smooth muscle within the SUFML can be quantified and there is variation among individuals. PMID- 12748474 TI - Irregular menses: an independent risk factor for gestational diabetes mellitus. AB - OBJECTIVE: Our purpose was to determine whether a history of irregular menses predicts gestational diabetes mellitus independently of traditional risk factors. STUDY DESIGN: We analyzed demographic characteristics, body mass index, and menstrual history of 85 pregnant women with gestational diabetes mellitus and compared them with 85 systematically selected control subjects who were matched for age, race, and delivery year. Subjects with pregestational diabetes mellitus, previous gestational diabetes mellitus, family history of diabetes mellitus, weight >200 pounds, previous macrosomic infants, or previous stillbirth were excluded. RESULTS: Demographic characteristics between case and control groups were similar. Mean body mass index was higher among cases (26.5 kg/m(2)) versus control subjects (24.5 kg/m(2), P =.004). Irregular cycles were more prevalent in the cases (24% vs 7%, P =.006). With the use of body mass index as a stratification factor, menstrual irregularity maintained a strong association with gestational diabetes mellitus (P =.014). CONCLUSION: A history of irregular menstrual cycles was a significant independent predictor of gestational diabetes mellitus. If selective screening is implemented for gestational diabetes mellitus, such history should be considered in the decision of whom to test. PMID- 12748476 TI - CA 125 levels in the preoperative assessment of advanced-stage uterine cancer. AB - OBJECTIVE: The purpose of this study was too evaluate preoperative levels of CA 125 in for the prediction of advanced uterine cancer. STUDY DESIGN: We conducted a retrospective analysis of the correlation of preoperative CA 125 with grade, depth of invasion, lymph vascular space involvement, lymph node status, and stage. RESULTS: High CA 125 levels correlated with advanced-stage (P <.0001) and positive (P <.0001) lymph node status. High levels of CA 125 also correlated with the deepest myometrial invasion, the presence of lymph vascular space involvement, and the highest grade. Receiver-operator characteristic curves demonstrated that depth of invasion, lymph vascular space involvement, and grade accurately predicted advanced-stage disease 73%, 77% and 80% of the time, respectively. CA 125 levels, however, correctly predicted advanced stage 94% of the time. The sensitivity and specificity of a CA 125 cutoff level of 37 IU/mL were 95% and 90%, respectively, with a positive predictive value of 78% and a negative predictive value of 97%. CONCLUSION: CA 125 appears to be a significant independent predictor of positive lymph node status and the extrauterine spread of disease. PMID- 12748475 TI - Predictors of success with methotrexate treatment of tubal ectopic pregnancy at Grady Memorial Hospital. AB - OBJECTIVE: The purpose of this study was to determine our institutional success rate with single-dose intramuscular methotrexate therapy for the treatment of ectopic pregnancy and to identify predictors of treatment outcome. STUDY DESIGN: This was a retrospective review of consecutive patients who were treated with methotrexate from January 2000 until April 2002. Successful treatment was defined as the resolution of the ectopic pregnancy without surgical intervention. RESULTS: Our overall success rate was 85% (69/81 patients). The median pretreatment serum beta-human chorionic gonadotropin level was lower in those women in whom treatment was successful compared with those women with treatment failure (793 vs 3804 mIU/mL, P <.002). The presence of a yolk sac that was visualized on vaginal ultrasonography was the only significant predictor of treatment failure (adjusted odds ratio, 19.3; P <.002). CONCLUSION: Our institutional success rate (85%) with methotrexate for treatment of tubal ectopic pregnancy is comparable with other published results. However, a finding unique to this study was that visualization of a yolk sac was a risk factor for failure. PMID- 12748477 TI - Effect of sleep deprivation on medical resident and student cognitive function: A prospective study. AB - OBJECTIVE: The purpose of this study was to determine whether cognitive function test results decrease after a typical night of on-call duty, on the basis of the amount of sleep that is obtained. STUDY DESIGN: Two standard cognitive functions tests (the Grooved Pegboard and California Verbal Learning Test II) were administered to the 30 participants who were members of the obstetrics and gynecology house staff and to medical students at Emory University School of Medicine. Each test was administered twice, before and after on-call duty. The data were analyzed with a paired two-tailed Student t test. RESULTS: For the California Verbal Learning Test II, trial 5 showed a decrease in score of 0.8 (P =.05) after on-call duty. Trials 1 through 5 showed a decrease in score of 3.5 (P =.01). CONCLUSION: This finding suggests that cognitive function test scores do decrease after on-call duty. Scrutiny of this issue by the Accreditation Council of Graduate Medical Education is justified, and the issue should continue to be evaluated. Our finding suggests that a change in our current educational structure is warranted. PMID- 12748478 TI - Patient satisfaction and the impact of written material about postpartum contraceptive decisions. AB - OBJECTIVE: The purpose of this study was to evaluate the impact of written educational material about patient satisfaction and decision-making regarding postpartum contraception. STUDY DESIGN: Baseline patient satisfaction and effectiveness of contraceptive counseling on a postpartum unit was evaluated with the use of a self-administered questionnaire. Data were compared with patients who received additional comprehensive written educational material during their postpartum hospitalization. RESULTS: A total of 109 women participated in the study (control subjects, 53; intervention group, 56). Demographics and patient satisfaction with contraceptive counseling were similar between groups. Thirty four percent of the control subjects reported having received some sort of written information; all women in the intervention group received a standardized comprehensive booklet of information during their postpartum stay (P <.01). Among the women who received written information, the intervention group was more likely to state that written material contributed to their ultimate choice in birth control (P <.01). CONCLUSION: The postpartum distribution of written material about contraceptive options increases a woman's ability to make an informed decision regarding birth control. PMID- 12748479 TI - Frequency and predictors for postpartum fecal incontinence. AB - OBJECTIVE: The goal of this study was to describe the frequency of postpartum fecal incontinence and to examine associated delivery events in a group of women participating in a validation study of a postpartum fecal incontinence survey. STUDY DESIGN: A self-administered survey of fecal incontinence symptoms and delivery events was administered to 50 women seen for their 6-week postpartum appointments. An analysis of association between postpartum fecal incontinence and individual delivery characteristics was performed. RESULTS: The frequency of at least one fecal incontinence symptom post partum was 38%. Specifically, 24% of women reported incontinence to flatus and 10% incontinence to stool. Vaginal delivery and perineal laceration were associated with an increased risk of reporting at least one symptom. Neither infant birth weight nor parity was associated with an increased risk of postpartum fecal incontinence. Cesarean delivery was associated with a lower frequency of overall fecal incontinence symptoms. CONCLUSION: The frequency of postpartum fecal incontinence was higher than previously reported. Similar to other studies, perineal laceration, especially extension into the anal sphincter, was an important risk factor for postpartum fecal incontinence. PMID- 12748480 TI - Estrogen increases inducible nitric oxide synthase gene expression. AB - OBJECTIVE: The goal of this study was to determine whether estrogen increases the expression of the inducible nitric oxide synthase gene. STUDY DESIGN: An inducible nitric oxide synthase fusion gene was created with its promoter and the reporter gene, luciferase. COS cells were transfected transiently with the fusion gene and cotransfected with an estrogen receptor-alpha expression plasmid to ensure the presence of an estrogen receptor. Cells were then exposed to estradiol (1 nmol/L and 10 nmol/L) or a cytokine mix that consisting of tumor necrosis factor-alpha, interleukin-1beta, and interferon gamma. Gene expression was measured in relative light units. RESULTS: Estradiol increased the expression of inducible nitric oxide synthase by an average of 31.2% in the COS cells that were cotransfected with estrogen receptor compared with -10.4% in cells without estrogen receptor (P =.006). CONCLUSION: Inducible nitric oxide synthase expression was increased with the addition of estrogen. These data support previous studies that demonstrated the inflammatory effects of estrogen and provides further insight into the mechanism by which estrogen might have an impact on the cardiovascular system. PMID- 12748481 TI - Expression of placental low-density lipoprotein receptor-related protein and scavenger receptors AI/AII transcripts in the baboon. AB - OBJECTIVE: This study was undertaken to assess potential secondary lipoprotein dependent mechanisms, scavenger receptor AI and AII (SR-AI and SR-AII), and low density lipoprotein (LDL) receptor-related protein (LRP) in the baboon (Papio sp) placenta. STUDY DESIGN: Baboon placental villous tissue and enriched fractions of syncytiotrophoblast cells were collected at early (approximately day 60, n = 3), mid (approximately day 100, n = 4), and late (approximately day 160, n = 4) pregnancy (term = approximately 184 days). The abundance of messenger RNA (mRNA) transcripts for SR-AI, SR-AII, and LRP were determined relative to constitutively expressed, glyceraldehyde-3-phosphate-dehydrogenase (GAPDH). RESULTS: No quantitative differences (P >.05) in SR-AI, SR-AII, or LRP mRNAs were noted in respect to stage of gestation. However, SR-AII/GAPDH mRNA ratios in syncytiotrophoblast cells (0.38 +/- 0.09, n = 11) were significantly (P <.05) higher than in villous tissue (0.15 +/- 0.04, n = 11), whereas LRP/GAPDH mRNA ratios were higher in villous tissue (0.43 +/- 0.10, n = 11) than in syncytiotrophoblast cells (0.18 +/- 0.02, n = 11). CONCLUSION: SR-AII mRNA in baboon placenta appears to be expressed, in greatest abundance, in the endocrinologically active syncytiotrophoblast. PMID- 12748482 TI - Postdates pregnancy in an indigent population: the financial burden. AB - OBJECTIVE: The objective of this study was to determine the comparative financial burden of twice-weekly fetal testing from 41 weeks of gestation until delivery, as compared with early dating ultrasound evaluation in an indigent population. STUDY DESIGN: All women who were seen for antepartum testing for postdating pregnancy at Lyndon Baines Johnson Hospital were enrolled. Patient age, parity, gestational age at initiation of prenatal care, the number of prenatal visits, gestational age at first ultrasound scan, and the number of biophysical profiles that were performed before delivery were recorded. The labor and delivery database was searched for all deliveries at >41 weeks of gestation. The charge for a single ultrasound scan at <20 weeks of gestation was compared with twice weekly testing in the population as a whole with the use of three strategies (no dating ultrasound scans and biophysical profiles until delivery, routine dating ultrasound scan and routine induction at 41 weeks of gestation, and current practice at our institution). RESULTS: One hundred twenty-seven subjects with postdated pregnancy were enrolled (mean age, 25.2 years; median parity, 0 [range, 0-6]). The mean gestational age at the initiation of prenatal care was 21.2 +/- 10.5 weeks. Forty-seven women (38.0%) initiated care at <20 weeks. The mean number of biophysical profiles performed before delivery was 1.5 +/- 1.34; the mean gestational age at delivery was 42.1 +/- 0.87 weeks (spontaneous labor, 39.6%; induced labor, 40.4%). The charge for a biophysical profile is $492.90 US dollars and $551.00 US dollars for a 20-week ultrasound scan; there is no difference in the charge for induced or spontaneous labor. During the 4-month study period, 1638 patients were delivered at our hospital; 341 patients were delivered at >41 weeks of gestation. The estimated financial burden of antenatal testing of 341 patients from 41 weeks to delivery was calculated to be $252,118 US dollars, compared with $902,538 US dollars for a single ultrasound scan at 20 weeks for the entire population of 1638 patients. The estimated financial burden of current practice (10% of patients with no prenatal care, 38% of patients with examination at <20 weeks who were eligible for dating ultrasound scanning, and 37% of patients with examination for postdate testing) was $402,457 US dollars. CONCLUSION: Patients who were seen for postdate antepartum testing in an indigent population lack early initiation of prenatal care and early ultrasound scans. Because on average only 1.5 biophysical profiles are performed per patient before delivery, routine early ultrasound scanning and routine induction at 41 weeks of gestation would add considerable financial burden to the system. PMID- 12748484 TI - The usefulness of the TDx/TDxFLx fetal lung maturity II assay in the initial evaluation of fetal lung maturity. AB - OBJECTIVE: This study was undertaken to evaluate the usefulness of the TDx/TDxFLx Fetal Lung Maturity II (FLM II) assay (Abbot Laboratories, Abbot Park, Ill) in the initial evaluation of fetal lung maturity. STUDY DESIGN: One hundred twenty seven women who underwent amniotic fluid collection through amniocentesis or vaginal pool for fetal lung maturity studies were evaluated for inclusion. The uncontaminated amniotic fluid specimens were analyzed within 24 hours with the use of the TDx/TDxFLx FLM II assay with values 55 mg/g or greater considered mature, values between 40 and 54 mg/g considered intermediate, and values 39 mg/g or less considered immature. Eighty-two women with singleton gestations who were delivered within 72 hours were included in the final analysis. We compared the TDX/TDxFLx FLM II test with the presence of neonatal respiratory distress syndrome (RDS). RESULTS: Seven of the 82 (8.5%) neonates had RDS develop. None of the 60 neonates with mature values had RDS develop. Two of 10 (20%) neonates with intermediate values and 5 of 12 (42%) neonates with immature values had RDS develop. With the use of a cutoff value of 55 mg/g, the TDx/TDxFLx FLM II assay had sensitivity and a negative predictive value of 100%, a specificity of 80%, and a positive predictive value of 31.8%. CONCLUSION: The TDx/TDxFLx FLM II assay is a useful tool in determining fetal lung maturity by using a cutoff value of 55 mg/g. PMID- 12748483 TI - Screening for postpartum depression in an inner-city population. AB - OBJECTIVE: This study was undertaken to determine the prevalence of positive screens for postpartum depression by using the Edinburgh Postnatal Depression Scale (EPDS) in an inner-city population. STUDY DESIGN: At their postpartum visits, women who were seen in two inner-city practices between February 20 and April 30, 2002, self-administered the EPDS. A threshold of 10 or more points was selected as a positive screen for postpartum depression. If the patient responded that she had thoughts of harming herself or others, she was referred immediately for social work or psychiatric evaluation. Otherwise, providers were blinded to the results of the screen. RESULTS: Of the 443 patients who were scheduled for postpartum visits during this time period, 285 (64%) patients presented for their postpartum visit. One hundred twenty-one (42%) of these patients participated in the study and 1 patient did not complete the questionnaire. Twenty-seven (22%) of the 121 patients had a positive screen for postpartum depression. Eight patients (6.6%) reported experiencing suicidal ideation within the previous week. Without use of the screening tool, providers identified 16 (13%) patients who they thought were at risk for postpartum depression. Of those 16 patients, 14 had positive screens on the EPDS. Eight patients eventually had clinical depression diagnosed. There was a significant relationship between women with elevated scores on the EPDS and a personal history of depression (P <.0006), history of prior treatment for depression (P <.0001), and feeling depressed during the pregnancy (P <.0001). Providers failed to comment on the patient's mood in 34 (28%) of the 121 patients. CONCLUSION: A larger than expected number of women had a positive screen for postpartum depression in our population. This unexpected finding suggests the need for more resources to be directed toward the research, identification, and treatment of postpartum depression in inner-city populations. PMID- 12748485 TI - Complications associated with peripherally inserted central catheter use during pregnancy. AB - OBJECTIVE: The purpose of this study was to evaluate the complications of peripherally inserted central catheter use in an obstetrics population at a single institution over a 5-year period. STUDY DESIGN: We conducted a descriptive retrospective review of all obstetrics patients with peripherally inserted central catheter insertion during the antepartum period at Good Samaritan Regional Medical Center, from January 1, 1997, through September 30, 2001. RESULTS: During the 5 years of the study, complete data regarding the primary admission diagnoses for peripherally inserted central catheter placement and associated complications were available for 52 cases. An associated complication rate of 50% (n = 26 cases) was noted. The complications included culture-proved line infection (n = 9/52 complications, 17%), presumed line infection (n = 6/52 complications, 12%), cellulitis (n = 4/52 complications, 8%), mechanical line failure (n = 4/52 complications, 8%), pain that required line discontinuation (n = 2/52 complications, 4%), and superficial thrombophlebitis (n = 1/52 complications, 2%). A chi(2) analysis was performed, and preterm labor may be associated with an increased risk for peripherally inserted central catheter complication (P =.012). CONCLUSION: Patients who receive a peripherally inserted central catheter during pregnancy are at significant risk for infectious complications. Alternative treatment approaches are suggested, where appropriate. PMID- 12748486 TI - Is placenta previa more common in multiple gestations? AB - OBJECTIVE: This study was undertaken to compare the occurrence of placenta previa at delivery between singleton and multiple gestations. STUDY DESIGN: A historical cohort study was performed of placenta previa cases at Good Samaritan Regional Medical Center, Phoenix, Ariz, from January 1, 1997, to December 31, 2000. RESULTS: During the study, 29,268 deliveries (28,372 singleton and 896 multiple gestations) occurred at Good Samaritan Regional Medical Center. Placenta previa complicated 51 (0.18%) singleton gestations and 4 (0.46%) multiple gestations (P =.09). Singleton and multiple gestations with placenta previa were similar with respect to maternal demographic variables, prior uterine surgery (excluding cesarean section), and gestational age at delivery. Multiple gestations with placenta previa were more likely to have had a prior cesarean section (P <.001). There were no differences between the singleton and multiple gestations with placenta previa and need for cesarean hysterectomy and blood transfusion. CONCLUSION: The occurrence and complications of placenta previa do not differ between singleton and multiple gestations. PMID- 12748487 TI - The association of fetal heart rate patterns with nucleated red blood cell counts at birth. AB - OBJECTIVE: The purpose of this study was to evaluate the relationship between fetal heart rate patterns and fetal nucleated red blood cell counts. STUDY DESIGN: Data were collected prospectively from March through September 2000. Umbilical cord blood was used for nucleated red blood cell analysis. The fetal heart rate pattern was analyzed for reactivity; presence, duration, and type of decelerations; and time interval between the last acceleration and delivery. RESULTS: Two hundred seventy-nine singleton term pregnancies formed the study population, 67% of which were delivered vaginally. The median nucleated red blood cells per 100 white blood cells was 7 (range, 0-158). The univariate analysis indicated that nucleated red blood cells are correlated significantly with abnormal fetal heart rate patterns, time interval between the last acceleration and delivery, fetal growth restriction, the presence of meconium, and a 1-minute Apgar score of < or =7. However, the stepwise regression analysis identified the time interval between the last acceleration and delivery as the only variable that independently predicted elevated nucleated red blood cell count at birth (P <.0001, R (2) = 0.26). CONCLUSION: Earlier studies have demonstrated an association between adverse perinatal outcome and elevated nucleated red blood cell count. In view of the high false-positive rate that is associated with the prediction of adverse perinatal outcome by fetal heart rate abnormalities, our results support previous studies that indicate that the presence of fetal heart rate accelerations is a reliable predictor of the nonhypoxic fetus. PMID- 12748488 TI - Perceived quality of life in pregnant adolescent girls. AB - OBJECTIVE: The purpose of this study was to describe the perceived quality of life and functional status of pregnant adolescent girls. STUDY DESIGN: The Medical Outcomes Survey-Short Form 36, version 2, is a multi-item scale that measures a subject's perception of eight health components. These questionnaires and demographic surveys were completed by teen patients aged 14 to 18 years during antenatal visits to a university-based resident continuity clinic. Mean scores (0 points indicates the poorest perceived health and functional status, 100 points indicates the highest perceived health and functional status) were then compared with mean scores for women in the general population of all ages and, specifically, women aged 18 to 24 years. RESULTS: Forty-two pregnant adolescent girls participated (median age, 16 years; median education level, 10th grade). Ninety-two percent of the girls belonged to a racial-ethnic minority, and all qualified for Medicaid. Pregnant adolescent girls had slightly lower overall scores for all components of perceived quality of life, with the exception of the vitality component. Scores were significantly lower than normative scores (by >1 SD) for only the physical functioning component. CONCLUSION: There appears to be little change in perceived quality of life in pregnant adolescent girls. This information may be useful in the provision of comprehensive care to pregnant adolescent girls. PMID- 12748489 TI - Interleukin-18 in the plasma of women with preeclampsia. AB - OBJECTIVE: Interleukin-18 (IL-18) is a proinflammatory cytokine capable of stimulating interferon gamma and tumor necrosis factor-alpha production. Our purpose was to determine whether abnormal levels of IL-18 in maternal plasma correlate with the diagnosis of preeclampsia. STUDY DESIGN: A case control study design was used to enroll 61 patients: controls (n = 31) and preeclamptic women (n = 30). A standard enzyme-linked immunosorbent assay measured plasma IL-18. Statistical methods included Student t tests and chi(2) tests. RESULTS: Mean IL 18 levels were lower in preeclampsia than in controls (185 +/- 74 pg/mL vs 224 +/ 75 pg/mL, P =.053). Administration of betamethasone (BMZ) and/or hydralazine (HYD) was significantly associated with a lower IL-18 compared with controls (159 +/- 50 pg/mL vs 224 +/- 75 pg/mL, P =.002). After women who received BMZ or HYD were excluded, levels of IL-18 in preeclampsia were similar to those of controls (213 +/- 87 pg/mL, P =.69). There was no association between gestational age and IL-18. CONCLUSION: Lower IL-18 was associated with administration of either BMZ or HYD. After patients receiving these medications were excluded, IL-18 levels were similar in controls and preeclamptic women. IL-18 was not associated with gestational age. PMID- 12748490 TI - Infant mortality, low birth weight, and prematurity among Hispanic, white, and African American women in North Carolina. AB - OBJECTIVES: The study was undertaken to compare Hispanic birth outcomes with those of white and African American women in North Carolina and to examine variables associated with adverse birth outcomes among Hispanic women. STUDY DESIGN: Retrospective comparison of birth outcomes by ethnicity/race, from linked birth/infant death certificates in North Carolina (1993-1997) was conducted. Multivariate, binary logistic regression and chi(2) analysis were used to examine relationships between available medical and sociodemographic index values and composite birth outcomes among Hispanic women. RESULTS: Infant mortality rates were lowest among Hispanic women. Low birth weight and prematurity rates were similar to those of white women and lower than those of African American women. Variables significantly related to healthy composite birth outcomes among Hispanic women included higher education, no preterm delivery history, prenatal care, marriage, and no daily tobacco use. CONCLUSION: Hispanic birth outcomes in North Carolina were better than those of African American women and similar to those of white women, despite use of prenatal care and socioeconomic characteristics similar to African American women. PMID- 12748491 TI - Sperm exposure and development of preeclampsia. AB - OBJECTIVE: Length of sperm exposure has been proposed to influence the risk of preeclampsia. The main objective was to determine the relationship between extent of exposure to sperm, both before and during pregnancy, and the risk of preeclampsia. STUDY DESIGN: A case-control design was used where women with preeclampsia (cases) were matched with two women without preeclampsia (controls) by age and parity. Data were analyzed by Student t test, chi(2) test, and logistic regression analysis. RESULTS: A total of 113 cases were compared with 226 controls. Women with a short period of cohabitation (<4 months) who used barrier methods for contraception had a substantially elevated risk for development of preeclampsia compared with women with more than 12 months of cohabitation before conception (odds ratio 17.1, P =.004). CONCLUSION: Fewer than 4 months of cohabitation among users of barrier methods for contraception is associated with a significantly increased risk for preeclampsia. PMID- 12748492 TI - The impact of urodynamic stress incontinence and detrusor overactivity on marital relationship and sexual function. AB - OBJECTIVE: We assessed and compared the quality of life, marital relationship, and sexual function of women who had urodynamic stress incontinence (USI) or detrusor overactivity (DO) diagnosed. STUDY DESIGN: Women with the following urodynamic diagnoses, normal, USI, or DO, were compared by using the psychometric questionnaires: King's Health Questionnaire (KHQ, for quality of life), Dyadic Adjustment Scale (DAS, for marital relationship), and Derogatis Sexual Functioning Inventory (DSFI, for sexual function). RESULTS: Women with DO (n = 29) were younger than healthy women (n = 28) and women with USI (n = 36) (P <.05). Women with USI or DO had a poorer quality of life (P <.05), poorer marital relationship (P <.05), and less sexual satisfaction (P <.05), despite having no change in sexual drive (P >.05). Regression analyses showed that only sexual satisfaction and incontinence-related emotions were associated with marital relationship (P <.05). CONCLUSION: Marital relationship and sexual function were negatively affected in women who had USI or DO. PMID- 12748493 TI - Ultrasound assessment of endometrial thickness and endometrial polyps in women on hormonal replacement therapy. AB - OBJECTIVE: This study was performed to evaluate the thickness and the sonographic features of the endometrium in postmenopausal women on hormone replacement therapy (HRT). STUDY DESIGN: This prospective study was based on 483 vaginal ultrasound examinations performed in 238 consecutive postmenopausal women on HRT. RESULTS: Endometrial polyps were diagnosed by ultrasound in 16.3% of the patients; 43.6% were asymptomatic. In the nonpolyp group, a significant difference in endometrial thickness between sequential and continuous schemes was found. In sequential HRT, a three-layer ultrasound pattern of the endometrium was seen in 47% of the women in the estrogen-alone phase of cycle versus 11% in the estrogen-progestogen phase. CONCLUSION: Patients on HRT need other reference values for endometrial thickness compared with postmenopausal women without HRT. In women on sequential HRT, a transvaginal sonographic evaluation performed in the estrogen-alone phase of cycle may optimize the accuracy for focal lesion detection. The significance of the incidental finding of polyps in an important number of asymptomatic women on HRT remains unsettled. PMID- 12748494 TI - Persistent low levels of human chorionic gonadotropin: A premalignant gestational trophoblastic disease. AB - OBJECTIVE: This study was undertaken to evaluate the significance of persistent low-level human chorionic gonadotropin (hCG) titers (usually <50 IU/L) in the absence of clinical evidence of pregnancy or gestational trophoblastic disease. STUDY DESIGN: The USA hCG Reference Service consulted on 114 cases with persistent low levels of hCG; 51 had false-positive hCG results. The remaining 63 cases had real hCG results and are presented here. RESULTS: Antecedent gestational events included hydatidiform mole (27), pregnancy (35), and gestational trophoblastic neoplasm (1). Forty of the 63 (64%) cases received therapy, including chemotherapy (38), hysterectomy (2), or both (10). Despite treatment, in all cases, low hCG titers persisted. After 1 to 4.5 years of low titers, four women had a sudden rapid increase in hCG levels, and malignant disease was confirmed or clearly suggested (gestational trophoblastic neoplasm [3] and placental site trophoblastic tumor [1]). Invasive trophoblast antigen (ITA) is a marker of invasive cytotrophoblast cells. ITA was measured in 38 of the cases with persistent low hCG, in all cases ITA accounted for less than 25% of the hCG concentration. It was also determined in the 4 cases indicated with malignant disease, accounting for more than 80% of the hCG. CONCLUSION: The presence of persistent low-level hCG titers defines a subset of women with preinvasive or quiescent gestational trophoblastic disease. ITA effectively detected the presence or absence of invasive cells in these cases. The recommended management of the quiescent disease is close surveillance without therapy until malignant disease detected. PMID- 12748495 TI - Vaginoplasty with Interceed absorbable adhesion barrier for complete squamous epithelialization in vaginal agenesis. AB - OBJECTIVE: The purpose of this study was to present our experiences of an innovative surgical approach for vaginal agenesis with the use of Interceed absorbable adhesion barrier (Ethicon) to achieve a satisfactory neovagina. STUDY DESIGN: The current study involved 10 subjects who were diagnosed with vaginal agenesis. After the creation of a vaginal tunnel, a mold that had been wrapped with Interceed was placed in the neovagina. RESULTS: No operative and postoperative complications were encountered. The duration of the operation was < or =30 minutes, and blood loss was minimal. The postoperative hospital stay was only 2 days. Epithelialization of the neovagina was achieved 1 to 4 months after the operation, and all patients were satisfied with the outcome. The neovagina that was created with this procedure was not much different from the normal adult vagina as far as histologic and physiologic conditions are concerned. CONCLUSION: This innovative surgical procedure may be a potential alternative approach for the therapy of vaginal agenesis with the use of the absorbable adhesion barrier Interceed with excellent results. PMID- 12748496 TI - Combined use of magnetic resonance imaging, CA 125 assay, histologic type, and histologic grade in the prediction of lymph node metastasis in endometrial carcinoma. AB - OBJECTIVE: The aim of this study was to predict retroperitoneal lymph node metastasis during the preoperative examination of patients with endometrial carcinoma and to determine whether lymphadenectomy must be performed. STUDY DESIGN: This study was carried out on 214 patients with endometrial carcinoma. Preoperative evaluators were volume index, depth of myometrial invasion (as assessed by magnetic resonance imaging), serum CA 125 level, histologic type, and histologic grade. With the use of receiver operating characteristic curves, cutoff values of volume index and serum CA 125 levels were determined. The relationships of these evaluators with pelvic lymph node metastasis were investigated by multivariate analysis with a logistic regression model. The relationships of these evaluators with para-aortic lymph node metastasis were investigated in the same way. RESULTS: Histologic type, volume index, histologic grade, and serum CA 125 level were found to be independent risk factors for pelvic lymph node metastasis; serum CA 125 level and volume index were found to be independent risk factors for para-aortic lymph node metastasis. Among 110 cases with no risk factors for pelvic lymph node metastasis, pelvic lymph node metastasis was observed in 4 cases (3.6%). On the other hand, only 1 case of 128 cases (0.7%) with no risk factors for para-aortic lymph node metastasis actually had metastasis. CONCLUSION: Careful consideration of the possibility of the elimination of the requirement of retroperitoneal lymphadenectomy is needed in cases with no risk factors for lymph node metastasis. However, our results suggest that para-aortic lymphadenectomy may not be necessary in cases with no risk factors for para-aortic lymph node metastasis. PMID- 12748497 TI - Prevention of ureteral injuries in gynecologic surgery. AB - Pelvic surgery is the most common cause of iatrogenic ureteral injury. The majority of patients with ureteral injuries have no identifiable predisposing risk factors. A simple maneuver that has been taught successfully at our institution that facilitates the identification of the ureter is described. When injury is discovered during surgery, correction of the injury can be repaired with minimal risk of long-term sequelae. Postoperatively, patients with ureteral injury typically present with costovertebral angle tenderness, ileus, fever, and flank pain with a minimal rise in serum creatinine. To prevent ureteral injuries, the surgeon must have a thorough knowledge of the location of the ureter during various pelvic procedures and the specific regions where it is most susceptible to injury. PMID- 12748498 TI - Administration of raloxifene does not influence 24-hour ambulatory blood pressure of postmenopausal women with osteopenia: a double-blind placebo-controlled study. AB - OBJECTIVE: Because estrogens may decrease 24-hour blood pressure of postmenopausal women, we tested the effect of the selective estrogen receptor modulator raloxifene on ambulatory blood pressure. STUDY DESIGN: Postmenopausal women with osteopenia who were otherwise healthy were assigned randomly in a double blind-fashion to receive placebo (n = 16 women) or raloxifene (60 mg/d, n = 16 women). Before and after 4 months, the blood pressure of each woman was monitored every 30 minutes for 41 hours with the use of an ambulatory device. RESULTS: Similar to the placebo outcome, the raloxifene administration did not modify 24-hour daytime (7 AM -11 PM) and nighttime (11 PM -7 AM) blood pressure and heart rate values. Day-night differences and the 24-hour rhythmic variation of mean blood pressure, which was evaluated by cosinor analysis, were also not affected by placebo or by raloxifene. CONCLUSION: Raloxifene does not influence 24-hour blood pressure of postmenopausal women. These data are reassuring for the cardiovascular safety of the long-term raloxifene administration. PMID- 12748499 TI - Inhibitors that target protein kinases for the treatment of ovarian carcinoma. AB - OBJECTIVE: Ovarian cancer is the leading cause of death from gynecologic malignancies in the United States. In an attempt to develop drugs that suppress ovarian cancer cells, we examined the effect of selective inhibitors of protein tyrosine kinases-tyrphostins, which are likely to play a role in ovarian cancer cells. STUDY DESIGN: We examined the cellular and biochemical effects of tyrphostins AG1478, PP2, AGL2592, and AG490 from four different families on the ovarian carcinoma cell line OV1063. RESULTS: We found that the AG1478, PP2, AGL2592, and AG490 tyrphostins suppressed cell proliferation and altered cell cycle distribution of the OV1063 cells in a dose- and time-dependent manner. Immunoblotting analysis indicated that AG1478 effectively inhibited epidermal growth factor receptor autophosphorylation, that AG490 decreased the level of Jak2 and phosphorylated Stat3, and that PP2 decreased the level of pp60Src protein. AGL2592 decreased the level of constitutive activated epidermal growth factor receptor and pStat3, but its molecular targets have not been identified completely. CONCLUSION: The growth-arresting properties of these tyrphostins identify them as possible candidates for signal transduction therapy. PMID- 12748501 TI - Treatment of nonatypical and atypical endometrial hyperplasia with a levonorgestrel-releasing intrauterine system. AB - OBJECTIVE: A "frameless" intrauterine drug delivery system that releases 14 microg/d of levonorgestrel was used to treat nonatypical and atypical endometrial hyperplasia in 12 women. STUDY DESIGN: This noncomparative study had up to 3 to 4 years of follow-up. RESULTS: The cure rate was 100%, as confirmed by repeat endometrial biopsy. CONCLUSION: This method could be considered an alternative to hysterectomy. PMID- 12748500 TI - Orally and transdermally replaced estradiol improves endothelial function equally in middle-aged women after surgical menopause. AB - OBJECTIVE: Improvement in endothelial function may be an important mechanism by which estrogen replacement therapy protects postmenopausal women against coronary artery disease. We determined whether the vascular effects of estradiol depend on the route of administration. STUDY DESIGN: Six weeks after surgically induced menopause, 43 healthy women were assigned randomly to 28 weeks of treatment by either orally or transdermally replaced estradiol. Endothelium-dependent and endothelium-independent dilation were calculated with the use of the diameters of the brachial artery that were measured at rest by high resolution ultrasound scanning after reactive hyperemia and after sublingual glyceryl trinitrate. RESULTS: Endothelium-dependent dilation increased after oral estradiol replacement from 6% +/- 3.9% to 13.2% +/- 4.4% (P <.0001) and after transdermal estradiol replacement from 7% +/- 4.9% to 14.9% +/- 5.6%(P <.0001). Endothelium independent dilation did not change significantly in either group. The improvements in endothelium-dependent dilation after estrogen substitution were independent of the changes in blood lipids and lipoproteins. CONCLUSION: Both oral and transdermal long-term replacement of estradiol lead to improved endothelial function in healthy middle-aged women after surgically induced menopause. PMID- 12748502 TI - The effect of placenta previa on neonatal mortality: a population-based study in the United States, 1989 through 1997. AB - OBJECTIVE: The purpose of the study was to explore the associations of placenta previa with preterm delivery, growth restriction, and neonatal survival. STUDY DESIGN: A retrospective cohort study was performed of live births in the United States (1989-1991 and 1995-1997) that used the national linked birth/infant death records from 22,368,235 singleton pregnancies. The diagnosis of previa was restricted to those live births that were delivered (> or =24 weeks) by cesarean delivery. We evaluated gestational age and birth weight-specific risk of neonatal deaths (within the first 28 days) in relation to placenta previa. Fetal growth was assessed in centiles of birth weight (<3rd, 3rd-4th, 5th-9th, 10th-90th, and >90th centile), adjusted for gestational age. All analyses were adjusted for the confounding effects of the year of delivery, maternal age, gravidity, education, prenatal care, marital status, and race/ethnicity. RESULTS: Placenta previa was recorded in 2.8 per 1000 live births (n = 61,711). Neonatal mortality rate was 10.7 with previa, compared with 2.5 per 1,000 among other pregnancies (relative risk, 4.3; 95% confidence interval, 4.0,4.8). At 28 to 36 weeks, babies born to women with placenta previa weighed, on average, 210 g lower than babies born to women without placenta previa (P <.001). Compared with babies born to women without previa, the risk of death from placenta previa was lower among preterm babies (<37 weeks of gestation), with a crossover at 37 weeks where the mortality rate was higher for babies born to women with placenta previa than for babies born to women without placenta previa. This crossover also persisted in an analysis by birth weight and term births (delivered at > or =37 weeks of gestation). Mortality rates for term births were higher among babies born to women with placenta previa than among babies born women without placenta previa who were at the 10th to 90th centile (relative risk, 1.9; 95% confidence interval, 1.3, 2.8), and those at >90th centile (relative risk, 3.6; 95% confidence interval, 1.3, 9.6). Among preterm births, however, placenta previa was not associated with increased neonatal mortality by fetal growth centiles. CONCLUSION: The risk of neonatal mortality was higher for babies born to women with placenta previa than for babies born to women without placenta previa who were delivered at > or =37 weeks of gestation. Pregnancies that are diagnosed with placenta previa must be monitored carefully, especially as they approach term. PMID- 12748504 TI - Noninvasive tests to predict fetal anemia: a study comparing Doppler and ultrasound parameters. AB - OBJECTIVE: This study was undertaken to compare test characteristics of ultrasound and Doppler parameters in the prediction of fetal anemia in alloimmunized pregnancies. STUDY DESIGN: In a prospective cohort study, 16 nonhydropic fetuses with red blood cell alloimmunization were evaluated with ultrasound and Doppler imaging. Middle cerebral artery (MCA) peak systolic velocity, intrahepatic umbilical venous (IHUV) maximum velocity, liver length, and spleen perimeter were measured. Results before first fetal blood sampling (FBS) or delivery were analyzed. Fetal anemia was defined as hemoglobin deficit 5 SD or greater. Sensitivity and specificity were calculated. RESULTS: Six fetuses were anemic and required intrauterine transfusion, and 10 were not severely anemic at birth. MCA Doppler imaging was the best predictor of fetal anemia (100%), followed by IHUV (83%). Sensitivity was low for spleen perimeter (66%) and liver length (33%). CONCLUSION: Doppler evaluation of MCA peak systolic velocity is better than IHUV maximum velocity, liver, or spleen size in the prediction of fetal anemia in red blood cell alloimmunization. PMID- 12748503 TI - Placenta previa: neonatal death after live births in the United States. AB - OBJECTIVE: The purpose of this study was to describe neonatal mortality rates among live births that were complicated by placenta previa in the United States. STUDY DESIGN: This was a population-based retrospective cohort study of 1997 United States singleton live births. Neonatal deaths among pregnancies that were complicated by placenta previa were compared with deaths among pregnancies with no placenta previa. Adjusted and unadjusted hazard ratios were generated from a proportional hazards regression model. RESULTS: Of 3,773,369 live births, 9656 were complicated by placenta previa (2.6 cases per 1000). Among cases of placenta previa, 114 neonatal deaths occurred (11.8 per 1000) versus 14951 (4 per 1000) among non-placenta previa neonates (P <.0001). The adjusted relative risk of death was three times higher among placenta previa neonates (hazard ratio, 3.06; 95% CI, 2.40-3.94). Placenta previa-related death was mediated through preterm delivery rather than small for gestational age. CONCLUSION: Placenta previa triples the rate of neonatal mortality, which is mediated mainly through preterm birth. PMID- 12748506 TI - Intrauterine growth restriction is accompanied by decreased renal volume in the human fetus. AB - OBJECTIVE: Intrauterine growth-restricted fetuses are at risk for the development of adult hypertension and related cardiovascular diseases. Congenital oligonephropathy has been postulated as the primary mechanism. The objective of our study was to determine whether ultrasonically obtained in utero measurements of renal volume or renal artery Doppler blood flow differ between fetuses that are intrauterine growth restricted and fetuses that are not. STUDY DESIGN: The study population consisted of women who were referred for a prenatal ultrasound evaluation at a large community medical center. The women were divided into two groups: women with fetal biometry that was consistent with intrauterine growth restriction and women with biometry within normal range. Information was collected on maternal demographics and other factors known to affect fetal growth. We performed detailed fetal renal anthropomorphic and Doppler blood flow measurements in addition to standard fetal biometric measurements on all patients, specifically comparing renal volume and renal artery flow data between the two groups. RESULTS: No differences were observed in maternal age, race, parity, or fetal gestational age. Renal volume in the intrauterine growth restricted fetuses was 31% (95% CI, 20%-40%), which was less than that in the group of fetuses that were not intrauterine growth restricted after an adjustment was made for gestational age. The ratio of renal volume to estimated fetal weight was 15% (95% CI, 1%-26%), which was less than the same ratio in the fetuses that were not intrauterine growth restricted. There were no differences seen in the renal artery Doppler measurements. CONCLUSION: Intrauterine growth restriction appears to be associated with a decrease in fetal renal volume. Because renal volume is a likely proxy for nephron number, this study supports the hypothesis that intrauterine growth restriction may be linked to congenital oligonephropathy and potentially to hypertension in later life. PMID- 12748505 TI - Medical abortion at 64 to 91 days of gestation: a review of 483 consecutive cases. AB - OBJECTIVE: The purpose of this study was to assess the uptake and outcome of medical abortion in the late first trimester of pregnancy. STUDY DESIGN: We conducted a review of the cases of 483 consecutive women in a university hospital who underwent medical abortion at 64 to 91 days of gestation and who used mifepristone that was followed 36 to 48 hours later by repeated doses of misoprostol. RESULTS: A total of 891 abortions were carried out at 64 to 91 days of gestation from October 2000 to April 2002; of these, 483 cases (54.2%) were undertaken medically. Complete abortion occurred in 458 cases (94.8%). Efficacy decreased with advancing gestational age. Surgical evacuation was carried out in 1 woman (0.9%) at 9 to 10 weeks of gestation, in 8 women (5.3%) at 10 to 11 weeks of gestation, in 7 women (6.2%) at 11 to 12 weeks of gestation, and in 9 women (7.9%) at 12 to 13 weeks of gestation. Indications for surgery included ongoing pregnancy in 8 cases (1.7%), missed abortion in 3 cases (0.6%), incomplete abortion in 13 cases (2.7%), and emergency curettage for bleeding in 1 case (0.2%). The mean number of misoprostol doses used was 2.3; of those women who had a complete abortion, 152 women (32.6%) aborted within 4 hours of receiving the misoprostol. The mean induction to abortion interval was 5.5 hours; most cases (452; 93.6%) were treated as day cases. CONCLUSION: Medical abortion between 64 and 91 days of gestation is effective and has a high uptake. Consideration should be given to extending the availability and choice of medical abortion to women in this gestational group. PMID- 12748507 TI - Leptin concentrations in maternal serum and cord blood in diabetic and nondiabetic pregnancy. AB - OBJECTIVE: The purpose of this study was to examine the relationships between maternal and cord leptin concentrations, maternal and neonatal outcomes, and measures of glycemic control in diabetic and nondiabetic pregnancy. STUDY DESIGN: This was a prospective study of 60 type 1 diabetic and 50 nondiabetic pregnancies in a university teaching hospital. Serum leptin and hemoglobin A(1c) were measured serially throughout pregnancy; leptin, insulin, insulin-like growth factor-1, and C-peptide in venous cord blood were measured at delivery. Leptin was measured with the use of enzyme-linked immunosorbent assay. Data were analyzed with specific computer software. RESULTS: Maternal leptin levels correlated with cord leptin levels in the nondiabetic group only. Cord leptin levels correlated with cord C-peptide, cord insulin-like growth factor-1, birth weight, birth weight corrected for gestational age, and neonatal anthropometry in both groups and with hemoglobin A(1c) in the diabetic group only. Cord leptin levels increased significantly with increasing birth weight corrected for gestational age but remained significantly higher at all birth weights in the diabetic group. CONCLUSION: There are strong associations between cord leptin levels and other measures of fetal growth in both groups and with glycemic control in the diabetic group. PMID- 12748508 TI - Stage-based treatment of twin-twin transfusion syndrome. AB - OBJECTIVE: The purpose of this study was to compare the outcomes of patients with twin-twin transfusion syndrome who were treated with either serial amniocentesis or selective laser photocoagulation of communicating vessels according to disease severity (stage). STUDY DESIGN: Centers that were experienced in the treatment of twin-twin transfusion syndrome were invited to share stage-based perinatal outcome data. All patients met basic standard sonographic criteria for twin-twin transfusion syndrome (polyhydramnios maximum vertical pocket, > or =8 cm; oligohydramnios maximum vertical pocket, < or =2 cm). Gestational age at first treatment was <27 weeks of gestation. RESULTS: Three centers submitted stage based data, for a total of 173 patients (serial amniocentesis, 78 patients from all 3 centers) and 95 selective laser photocoagulation of communicating vessels (1 center). The distribution of patients by stage was similar in the two groups. Successful pregnancy outcome (at least 1 surviving infant) was correlated inversely with stage in the serial amniocentesis but not in the selective laser photocoagulation of communicating vessels group and was significantly lower in the serial amniocentesis (66.7%) than in the selective laser photocoagulation of communicating vessels group (83.2%). Neurologic morbidity was related directly to stage in the serial amniocentesis group but not in the selective laser photocoagulation of communicating vessels group and was significantly higher in the serial amniocentesis (24.4%) than in the selective laser photocoagulation of communicating vessels (4.2%) group. Intact neurologic survival (at least 1 surviving infant without neurologic morbidity) was significantly lower in the serial amniocentesis group than in the selective laser photocoagulation of communicating vessel group (51.3% vs 78.9%), particularly in stage III and stage IV (23.5% vs 72.7% in stage IV). Patients who were treated with selective laser photocoagulation of communicating vessels were 2.4 times more likely to have at least one survivor than those treated with serial amniocentesis. CONCLUSION: Our study suggests a relationship between perinatal morbidity and mortality rates and stage in serial amniocentesis but not in selective laser photocoagulation of communicating vessel-treated twin-twin transfusion syndrome patients. These findings could be used to tailor the treatment of twin-twin transfusion syndrome. A clinical trial to confirm these results is being organized by our research groups. PMID- 12748509 TI - Intimate partner violence and adverse pregnancy outcomes: a population-based study. AB - OBJECTIVES: The purpose of this study was to measure the prevalence of exposure to intimate partner violence during pregnancy and to determine whether such exposure is associated with adverse pregnancy outcomes. STUDY DESIGN: We measured the prevalence of exposure to intimate partner violence and fear of a partner during pregnancy among 4750 residents of Vancouver, British Columbia, who gave birth between January 1999 and December 2000. We undertook a multivariate analysis to examine the associations with second- or third-trimester hemorrhage, preterm labor and delivery, intrauterine growth restriction, and perinatal death. RESULTS: We report a prevalence rate of 1.2% for exposure to physical violence by an intimate partner during pregnancy and 1.5% for fear of a partner. Physical violence was associated with an increased risk of antepartum hemorrhage (adjusted odds ratio [OR]: 3.79, 95% CI 1.38-10.40), intrauterine growth restriction (OR: 3.06, 95% CI 1.02-9.14), and perinatal death (OR: 8.06, 95% CI 1.42-45.63). Fear of a partner in the absence of physical violence was not associated with an elevated risk of adverse pregnancy outcomes. CONCLUSION: Our study confirms prior work reporting an association of physical abuse during pregnancy with intrauterine growth retardation and, in addition, reports an association with antepartum hemorrhage and perinatal death. PMID- 12748510 TI - Effect of maternal docosahexaenoic acid supplementation on postpartum depression and information processing. AB - OBJECTIVE: The purpose of this study was to determine the effect of docosahexaenoic acid supplementation on plasma phospholipid docosahexaenoic acid content and indices of depression and information processing for women who breast feed. STUDY DESIGN: Mothers who planned to breast-feed their infants were assigned randomly in a double-masked fashion to receive either docosahexaenoic acid (approximately 200 mg/d) or placebo for the first 4 months after the delivery. Major outcome variables included plasma phospholipid fatty acid patterns and scores on a self-rating questionnaire of current depression symptoms. A structured clinical interview of depression, scores on another self rating questionnaire of depression symptoms, and a laboratory measure of information processing were obtained in subgroups of the total population. RESULTS: Plasma phospholipid contents of docosahexaenoic acid at baseline were 3.15 +/- 0.78 and 3.31 +/- 0.70 (mg/dL of total fatty acids) in the docosahexaenoic acid and placebo groups, respectively. After 4 months, the plasma phospholipid docosahexaenoic acid content of the docosahexaenoic acid group was 8% higher (3.40 +/- 0.97 mg/dL), whereas that of the placebo group was 31% lower (2.27 +/- 0.87 mg/dL). Despite the higher plasma phospholipid docosahexaenoic acid content of the supplemented group after 4 months, there was no difference between groups in either self-rating or diagnostic measures of depression; information processing scores of the two groups also did not differ. CONCLUSION: Docosahexaenoic acid supplementation ( approximately 200 mg/d) for 4 months after the delivery prevented the usual decline in plasma phospholipid docosahexaenoic acid content of women who breastfeed but did not influence self-ratings of depression, diagnostic measures of depression, or information processing. PMID- 12748511 TI - Association between second-trimester isolated high maternal serum maternal serum human chorionic gonadotropin levels and obstetric complications in singleton and twin pregnancies. AB - OBJECTIVE: The purpose of this study was to examine the clinical significance of high maternal serum human chorionic gonadotropin levels in the second trimester in singleton and twin pregnancies within the Ontario maternal serum screening program. STUDY DESIGN: The study group comprised 564 women with singleton pregnancies with total maternal serum human chorionic gonadotropin levels of > or =4.0 multiples of the median (MoM) and serum marker alpha-fetoprotein levels of <2.0 MoM. The cases were matched with 1692 control subjects who had both serum marker alpha-fetoprotein levels and maternal serum human chorionic gonadotropin levels of <2.0 MoM. The second part of the study comprised 93 twin pregnancies with maternal serum human chorionic gonadotropin levels of > or =5.0 MoM and serum marker alpha-fetoprotein levels of <4.0 MoM; the control group (n = 1496) had serum marker alpha-fetoprotein levels of <4.0 MoM and maternal serum human chorionic gonadotropin levels of <5.0 MoM. The final part of the study included 25 women with extremely high maternal serum human chorionic gonadotropin levels (> or = 14;10 MoM). RESULTS: Of the singleton pregnancies with maternal serum human chorionic gonadotropin levels of > or = 14;4.0 MoM, 22.5% had severe adverse obstetric outcomes, compared with only 10.9% of the matched control population (P =.001). Women with markedly elevated maternal serum human chorionic gonadotropin levels had significantly increased risks of having spontaneous miscarriage, small-for-gestational-age infants, pregnancy-associated hypertensive disorder, and preterm delivery. Of the women with twin pregnancies with high maternal serum human chorionic gonadotropin levels (> or =5.0 MoM), 71% had at least one complication (such as miscarriage and preterm delivery) compared with 55.3% in the control group. Finally, 23 of 25 women with extremely high maternal serum human chorionic gonadotropin levels (> or = 14;10 MoM) had serious adverse outcomes (such as fetal abnormalities, pregnancy-associated hypertensive disorder, premature separation of placenta, intrauterine growth restriction, neonatal respiratory distress syndrome, and neonatal jaundice). CONCLUSION: Pregnancies with an elevated maternal serum human chorionic gonadotropin level are associated with adverse obstetric outcomes. Increased maternal and fetal surveillance is warranted in these pregnancies. PMID- 12748512 TI - The effect of isosorbide dinitrate, a donor of nitric oxide, on maternal cerebral blood flow in gestational hypertension and preeclampsia. AB - OBJECTIVE: The purpose of this study was to investigate the effect of a nitric oxide donor on cerebral perfusion pressure and other blood flow index values in the maternal middle cerebral and basilar arteries. STUDY DESIGN: Sublingual tablets of 5-mg isosorbide dinitrate were administered to 19 hypertensive pregnant patients. Doppler velocimetry of the middle cerebral and basilar arteries was obtained with a transcranial Doppler with the use of the transtemporal and suboccipital approach. RESULTS: The mean (+/-SEM) flow velocity in the middle cerebral artery significantly decreased from a baseline of 65.9 +/- 3.1 cm/s to 55.2 +/- 2.2 and 52.0 +/- 2.1 cm/s at 10 and 20 minutes (P <.0001). Resistance area product insignificantly increased from a baseline of 2.0 +/- 0.1 before isosorbide dinitrate to 2.19 +/- 0.11 at 20 minutes. No significant changes were observed in the resistance and pulsatility indices in the middle cerebral artery. The cerebral perfusion pressure did not change significantly after isosorbide dinitrate (84.5 +/- 7.3, 80.8 +/- 6.6, and 78.5 +/- 5.0 mm Hg at 0, 10, and 20 minutes, respectively) nor did the cerebral blood flow index. CONCLUSION: The results obtained demonstrate that cerebral perfusion pressure is unaltered by isosorbide dinitrate, despite significant changes in maternal blood pressure and in blood flow velocities in the middle cerebral artery. PMID- 12748513 TI - Screening for fetal well-being in a high-risk pregnant population comparing the nonstress test with umbilical artery Doppler velocimetry: a randomized controlled clinical trial. AB - OBJECTIVE: The purpose of this study was to evaluate the ability of two different modes of antepartum fetal testing to screen for the presence of peripartum morbidity, as measured by the cesarean delivery rate for fetal distress in labor. STUDY DESIGN: Over a 36-month period, all patients who were referred to the Fetal Assessment Unit at BC Women's Hospital because of a perceived increased fetal antepartum risk at a gestational age of > or =32 weeks of gestation were approached to participate in this study. Fetal surveillance of these women was allocated randomly to either umbilical artery Doppler ultrasound testing or nonstress testing as a screening test for fetal well-being. If either the umbilical artery Doppler testing or the nonstress testing was normal, patients were screened subsequently with the same technique, according to study protocol. When the Doppler study showed a systolic/diastolic ratio of >90th percentile or the nonstress testing was equivocal (ie, variable decelerations), an amniotic fluid index was performed, as an additional screening test. When the amniotic fluid index was abnormal (<5th percentile), induction and delivery were recommended. When the Doppler study showed absent or reversed diastolic blood flow or when the nonstress test result was abnormal, induction and delivery were recommended to the attending physician. Statistical comparisons between groups were performed with an unpaired t test for normally distributed continuous variables and chi(2) test for categoric variables. RESULTS: One thousand three hundred sixty patients were assigned randomly to groups in the study; 16 patients were lost to follow up. Six hundred forty-nine patients received Doppler testing and 691 received nonstress testing. The mean number of visits for the Doppler test and nonstress test groups was two versus two, respectively. The major indications for fetal assessment included postdates (43%), decreased fetal movement (22%), diabetes mellitus (11%), hypertension (10%), and intrauterine growth restriction (7%). The incidence of cesarean delivery for fetal distress was significantly lower in the Doppler group compared with the nonstress testing group (30 [4.6%] vs 60 [8.7%], respectively; P <.006). The greatest impact on the reduction in cesarean deliveries for fetal distress was seen in the subgroups in which the indication for testing was hypertension and suspected intrauterine growth restriction. CONCLUSION: Umbilical artery Doppler as a screening test for fetal well-being in a high-risk population was associated with a decreased incidence of cesarean delivery for fetal distress compared to the nonstress testing, with no increase in neonatal morbidity. PMID- 12748514 TI - Macrosomic births in the united states: determinants, outcomes, and proposed grades of risk. AB - OBJECTIVE: We describe maternal risk factors for macrosomia and assess birth weight categories to determine predictive thresholds of adverse outcomes. STUDY DESIGN: We analyzed linked live birth and infant death cohort files from 1995 to 1997 for the United States with the use of selected term (37-44 weeks of gestation) single live births to mothers who were US residents. We compared macrosomic infants (4000-4499 g, 4500-4999 g, and >5000 g infants) with a normosomic control group of infants who weighed 3000 to 3999 g. RESULTS: Maternal risk factors for macrosomia included nonsmoking, advanced age, married, diabetes mellitus, hypertension, and previous macrosomic infant or pregnancy loss. The risks of labor complications, birth injuries, and newborn morbidity rose with each gradation of macrosomic birth weight. Infant mortality rates increased significantly among infants weighing >5000 g. CONCLUSION: Although a definition of macrosomia as >4000 g (grade 1) may be useful for the identification of increased risks of labor and newborn complications, >4500 g (grade 2) may be more predictive of neonatal morbidity, and >5000 g (grade 3) may be a better indicator of infant mortality risk. PMID- 12748515 TI - Response of uterine and umbilical circulations to physical exercise. PMID- 12748517 TI - Much research is needed to provide fully informed consent about mode of delivery. PMID- 12748520 TI - Reducing operating costs. PMID- 12748521 TI - Pediatric anaphylaxis. PMID- 12748525 TI - Medical condition list and appropriate use of air medical transport. PMID- 12748526 TI - Life Flight Network celebrates 25 years of caring. PMID- 12748527 TI - Out of the box: Group rethinks ECMO transport for girl's sake. PMID- 12748528 TI - Care in consortiums. PMID- 12748529 TI - Presumption of death by air medical transport teams. AB - BACKGROUND: The purpose of this study was to investigate nationwide trends and factors influencing the determination of death practice by rotor-wing air medical transport programs. METHODS: A survey was mailed to all Association of Air Medical Service members concerning demographics, crew configuration, team leader, patient population, field death determination protocols, and other possible associated factors. All rotor-wing air medical transport programs that carry out scene transports were included. RESULTS: The most common field presumption criteria were no response to advanced cardiac life support (77%), no signs of life on scene (65%), and asystole in 2 EKG monitor leads (61%). The most frequent reasons cited not to presume a patient dead in the field were political issues (71%) and signs of life on scene (56%). Criteria other than medical condition that were considered in the decision to presume death were ground personnel input (55%) and program policy/medical control (39%). The following factors did not significantly affect the presumption rate: crew configuration, team leader, transport time, billing, and type of medical control. CONCLUSION: Medical criteria appear to determine presumption of death in the field. Nonmedical factors, such as billing, response, and transport times, do not affect this process. PMID- 12748531 TI - The airway pipeline: How do you know where your ETT is? PMID- 12748530 TI - Use of the autolaunch method of dispatching a helicopter. AB - INTRODUCTION: Autolaunch is a method of dispatching whereby the dispatcher can send the helicopter to a scene, as opposed to traditional request-driven dispatch. The purpose of this study was to investigate differences in patient outcomes when autolaunch was used. A 2-year period, July 1997 through June 1999, was studied. METHODS: A case control design was used. A retrospective chart review included 17 autolaunch and 16 traditionally dispatched patients. Patients were matched using Injury Severity Scores, Glasgow Coma Scale, and age. Eight matched pairs were used for statistical analysis. Three research questions were answered. RESULTS: The difference in time from accident to helicopter arrival was 3.64 minutes faster for autolaunch (statistical significance P =.336). Mortality data showed no statistical significance difference (P =.302). Intensive care unit (ICU) and hospital length of stays were both decreased with the use of autolaunch, although not statistically significantly. DISCUSSION: Sample size was small, making statistical significance difficult to achieve despite decreased length of stays and quicker time to the scene. CONCLUSION: Although statistical significance was not found with the use of autolaunch, patient outcomes still were improved by this method. Information provided could be used by helicopter programs considering implementing autolaunch. PMID- 12748535 TI - Count your fingers. More stuff on restraints. PMID- 12748532 TI - A case study of Brugada syndrome. PMID- 12748536 TI - Ambulance crashes: protect yourself and your patients. PMID- 12748537 TI - EMS: when it matters most. Spotlight on EMS Week 2003. PMID- 12748538 TI - All that glitters is not gold. PMID- 12748540 TI - Silent MCI. EMS agencies cope with unique obstacles to patient triage & treatment. PMID- 12748542 TI - The pelvic sheet wrap. Initial management of unstable fractures. PMID- 12748541 TI - EMS Marketing Made Easy: A 4-step plan to kick start your service's strategy-plus EMS experts from New York to Washington reveal their best marketing ideas. PMID- 12748543 TI - BLS-ALS EMT bridge. One city's unique way to transition BLS EMTs to an ALS system. PMID- 12748544 TI - Spirits in the sky. How non-traditional partnerships helped 4 areas creatively develop EMS helicopter services for their communities. PMID- 12748545 TI - A clean ride. Building EMS vehicles for easier cleaning & decontamination. PMID- 12748548 TI - Let's get physical. The importance of staying fit for EMS. PMID- 12748546 TI - Burn management. A comprehensive review of the epidemiology and treatment of burn victims. PMID- 12748549 TI - Devices & methodologies for capturing data. PMID- 12748552 TI - California organizes readiness for terrorism/MCI responses. PMID- 12748554 TI - Allergic rhinitis: broader disease effects and implications for management. AB - Allergic rhinitis is a burdensome disease for a significant part of the population in both adults and children. Poorly controlled allergic rhinitis can trigger exacerbations of asthma, sinusitis, and otitis media, diseases with which it shares common pathophysiologic elements. Consequently, early diagnosis and treatment should be a priority for patients and physicians, not only to control the symptoms of allergic rhinitis but also to improve the management of associated diseases. Several pharmacologic therapies can be considered in an armamentarium that includes antihistamines (intranasal and systemic), intranasal cromolyn, intranasal anticholinergic agents, intranasal steroids, systemic steroids, immunotherapy, and, most recently, leukotriene receptor antagonists. Often, combinations of these treatments are used to maximize control of refractory symptoms. PMID- 12748553 TI - Injection snoreplasty: extended follow-up and new objective data. AB - OBJECTIVE: Injection snoreplasty was recently introduced as a safe, effective, and minimally invasive treatment for primary snoring. Extended follow-up data (19 months) are presented from the initial pilot study cohort, and pretreatment/posttreatment objective data are presented on a new prospective patient cohort. STUDY DESIGN AND SETTING: The successfully treated patients from the initial pilot study (n = 25) were surveyed regarding their current snoring level, overall discomfort, and overall satisfaction. A new patient cohort with primary snoring (Respiratory Disturbance Index <10) was prospectively treated with injection snoreplasty. Snoring was objectively measured pretreatment and posttreatment using a take-home acoustical analysis device. RESULTS: Subjective success dropped from 92% to 75% at a mean of 19-month follow-up with a snoring relapse rate of 18%. Objective analysis of a new patient cohort (n = 17) confirmed statistically significant decreases in palatal flutter snoring and palatal loudness after injection. Preprocedure measurements were highly correlated to eventual treatment success. CONCLUSIONS AND SIGNIFICANCE: Success and snoring relapse rates of injection snoreplasty are similar to those of other current treatments. Objective analysis confirms the procedure is effective in reducing palatal flutter snoring. Preprocedure snoring analysis may predict patient response to palatal stiffening treatments. PMID- 12748555 TI - Caspase 3 activation in nasal capillary in patients with epistaxis. AB - OBJECTIVE: We investigated whether an apoptosis of nasal microvessels contributes to probable mechanism of the onset of epistaxis. STUDY DESIGN AND SETTING: Nasal septal mucosa of Little's areas taken from patients without (n = 19) and with (n = 26) epistaxis were examined. Active caspase-3 in the mucosa was detected according to the methods of immunohistochemistry and Western blotting. On Western blot analysis of the homogenates of the mucosa, we also sought probable signaling factors after caspase-3 activation. RESULTS: Marked activation of caspase-3 was detected in the capillaries and its neighboring muscle cells of Little's area from patients with epistaxis, and the activation was due to enhanced expression of procaspase-3 protein and progressive cleavage of the precursor. As a result of Western blotting of signaling factors, enhanced expressions of caspase-9 and Bax protein in the homogenates of Little's area in epistaxis group were found compared with those in control group. Increased levels of cytochrome c released into a cytosol were also detected in the capillaries in epistaxis group. CONCLUSION: In the present study, caspase-3 activation was found in the capillaries of Little's area from patients with epistaxis, suggesting that an apoptosis of capillaries may contribute to a mechanism of the onset of epistaxis. Moreover, alterations of some apoptotic factors such as caspase-9, Bax, and cytochrome c in the tissues demonstrated participation of mitochondrial disturbance in one of the apoptotic mechanisms. SIGNIFICANCE: Further explorations of the pathobiologic mechanism of capillary apoptosis can lead not only to an identification of risk factors in the onset of epistaxis but also to the development of medical therapy of epistaxis. PMID- 12748556 TI - The lateral crural J-flap repair of nasal valve collapse. AB - OBJECTIVES: Nasal valve collapse is a common cause of nasal airway obstruction. Although many techniques have been devised to correct both the functional and aesthetic aspects of this problem, none is uniformly successful. I propose that the true locus and solution to the problem lies lateral to the nasal valve angle in the fibroareolar tissue that connects the lateral aspect of the lateral crus to the bony pyriform aperture. Age-related or surgically induced weakening and loosening of these structurally important, fibroareolar connections result in a prolapse of the lateral aspect of the lateral crura away from the pyriform margin and toward the septum with a consequent narrowing of the nasal valve region. A new method of repair is described. METHODS: Bilateral marginal incisions are made following the curvature of the palpated caudal edge of the lower lateral cartilage starting at the dome region medially. The incision is extended laterally until the entire lateralmost aspect of the lateral crus is freed, generating a J-shaped chondrocutaneous flap that is medially and superiorly based. Following this, supraperichondrial dissection of the lateral crus is carried out, exposing the dome region and the upper lateral cartilage. The J-flap is then pulled caudally and laterally, and the excess overlapping tissue is evaluated. Two composite strips of vestibular skin and cartilage are resected: one parallel to the marginal incision and the second at the lateral edge of the J flap. The flap is transposed and sutured into position, pulling open the nasal valve angle. RESULTS: Preliminary results are presented for 18 patients who underwent 19 lateral crural J-flap repairs of nasal valve collapse. Nine patients underwent concomitant septal, sinus, or turbinate procedures. Two patients underwent unilateral J-flap repair. One patient required revision. Eighty-nine percent of patients reported markedly improved nasal airway patency and elimination of the subjective sensation of inspiratory collapse. Mean time to follow-up was 257 days. There has thus far been no noticeable decrement in benefit over time. CONCLUSION: This new technique for the treatment of nasal valve collapse may offer a simple and effective approach to the problem of nasal valve collapse with minimal morbidity and a high rate of success. The technique is based on a new view of the structural etiology of nasal valve collapse. PMID- 12748558 TI - Multivariate analysis of prognostic factors in T3 N0 laryngeal carcinoma treated with total laryngectomy. AB - OBJECTIVE: The aim of our study was to clarify the real importance of 16 possible prognostic factors analyzed retrospectively for the disease-free interval and survival of a total of 327 patients with T3 N0 M0 laryngeal carcinoma treated with total laryngectomy. METHODS: The role of each possible prognostic factor and their joint effect was explored by Cox proportional hazard survival analysis. Results and conclusions In a Cox univariate analysis for the whole group, tumor site, pattern of growth, tumor size, histologic grading, lymph node status, and the occurrence of postoperative complications were predictive of the risk of tumor recurrence. In univariate analysis, all these factors except for pattern of tumor growth, neck treatment and postoperative complications maintained their prognostic value when analyzed as predictors of survival; however, the loco regional control of disease was the most significant one. In a Cox multivariate analysis tumor size, histologic grading and postoperative complications had a significant impact on disease-free survival, whereas only histologic grading and loco-regional failure appeared to be prognostic with a significant decrease in overall survival. PMID- 12748557 TI - Kikuchi's disease: a review and analysis of 61 cases. AB - OBJECTIVE: Kikuchi's disease (KD), or histiocytic necrotizing lymphadenitis, is a rare self-limiting disorder that typically affects the cervical lymph nodes. It has occasionally been misdiagnosed as malignant lymphoma or another serious diseases; hence, clinicians should be made more aware of this disease. METHODS: From January 1986 to January 2001, a series of 61 patients who underwent a biopsy of a cervical lymph node and proved histologically to have KD were enrolled in this study. We retrospectively reviewed the clinical records and pathologic parameters of each patient. The patients were followed up from 6 months to 14.3 years (mean, 6.9 years). RESULTS: There were 34 women and 27 men (1.26:1 ratio; age range, 6 to 46 years; mean age, 21 years). The affected cervical lymph nodes were commonly located in the posterior cervical triangle (54 of 61, or 88.5%). Unilateral and bilateral cervical lymph nodes were affected in 54 and 7 patients, respectively. The dimensions of affected lymph nodes were commonly in the range of 0.5 to 4 cm (57 of 61, or 93.4%). In 2 patients, the size of the enlarged lymph nodes was greater than 6 cm. Leukopenia was observed in 14 patients (23%); fever, in 18 patients (29.5%). Five patients had a past history of tuberculosis. Two patients developed systemic lupus erythematosus, 1 month and 5 years later, respectively. The cervical lymphadenopathy usually resolved without any medical treatment within 6 months after definite diagnosis was made. In 3 patients, the enlarged lymph nodes eventually disappeared after more than 1 year. No recurrence has since been noted. CONCLUSION: The results of this study support the theory that KD is a self-limiting disorder that does not require any specific management. The female predominance was not as striking as in the studies performed in Western countries. We suggest that the patients with KD require a systemic survey and regular follow-up for several years; 2 of our patients developed systemic lupus erythematosus. An effective communication between the otolaryngologist and pathologist is needed because the clinical and pathologic characteristics of KD are essential in making an accurate diagnosis. PMID- 12748559 TI - Nonmelanoma cutaneous malignancy with regional metastasis. AB - OBJECTIVE AND STUDY DESIGN: We performed a retrospective study of 28 patients to evaluate the management and outcome of regional metastasis from nonmelanoma cutaneous malignancies. RESULTS: There were 25 squamous cell, 1 basal cell, and 2 eccrine carcinomas. The most common primary locations were the temple, ear, and cheek. Median interval from primary resection to regional metastasis was 9 months, and such were commonly of the parotid, level IB, and level IIA nodes. Twenty-four patients underwent parotidectomy and/or neck dissection; 19 patients, postoperative radiation. Ten patients (36%) died from the disease, 2 patients (7%) have persisting disease, 5 (18%) have died from intercurrent disease, and 11 (39%) have no evidence of disease (minimum follow-up, 22 months; median, 34 months). CONCLUSIONS AND SIGNIFICANCE: Nonmelanoma cutaneous malignancies with regional metastasis have a poor prognosis despite aggressive therapy. When high risk characteristics are detected, examination of the nearest "sentinel node," prophylactic lymphadenectomy, and/or regional irradiation might be justified; further study is warranted. PMID- 12748560 TI - Gene polymorphisms in detoxification enzymes as susceptibility factor for head and neck cancer? AB - OBJECTIVE: Impaired detoxification of carcinogens found in tobacco smoke appears to increase the risk for tobacco associated cancer. The objective of this study was to investigate concomitant polymorphisms in genes encoding for various detoxification enzymes in patients with head and neck squamous cell carcinoma (HNSCC). METHODS: In 187 patients with HNSCC and in 139 healthy control subjects, the polymorphisms of cytochrome P450 1A1 (CYP1A1), cytochrome P450 2D6 (CYP2D6), and glutathione S-transferase mu1 and Theta (GSTM1, GSTT1) were detected by polymerase chain reaction. RESULTS: No significant association were identified between CYP1A1 and CYP2D6 gene polymorphisms and HNSCC. Patients with laryngeal cancer revealed the GSTM1 null genotype more frequently than did the control subjects (P < 0.05). The coincidence of GSTM1 and GSTT1 null genotype was found twice as great in patients as in control subjects (P < 0.05). CONCLUSIONS: It is assumed that detoxification enzymes are functionally redundant and only the simultaneous deficiency of several detoxification enzymes increase the risk for HNSCC in alcohol- and tobacco-exposed individuals. PMID- 12748561 TI - Anterior skull base surgery without prophylactic airway diversion procedures. AB - OBJECTIVE: Although anterior skull base surgery has become a relatively safe and effective procedure, postoperative complications remain a serious problem. One of the most devastating complications of anterior skull base procedures is tension pneumocephalus (TP). In order to prevent TP, authors have recommended the use of prophylactic airway diversion procedures, such as prolonged endotracheal intubation or prophylactic tracheostomy. However, these procedures may mask neurologic deterioration, delay treatment, and prolong rehabilitation. The purpose of this study was to determine the need for airway diversion procedures in anterior skull base surgery. STUDY DESIGN: Eighty-five patients underwent anterior skull base operations through the subcranial approach without prophylactic airway diversion. Sixty-four patients underwent resection of tumors, 12 patients underwent repair of cerebrospinal fluid leak, 6 patients underwent surgery due to anterior skull base fungal infections, and 3 patients underwent anterior skull base reconstruction procedures. RESULTS: The complication rate of TP was 1.2% (1/85). This complication rate is similar to that previously reported for operations performed with airway diversion procedures. CONCLUSION: Prophylactic airway diversion procedures are unnecessary in routine anterior skull base operations. Airway diversion should be indicated only when factors that might predispose the patient to risk of TP have been identified (ie, chronic cough or obstructive pulmonary diseases). PMID- 12748562 TI - New endoscopic secondary tracheoesophageal voice prosthesis placement technique. AB - OBJECTIVE: The aim of this study was to present a new technique of secondary vocal prosthesis placement on an outpatient basis without general anesthesia by means of digestive endoscopy. METHODS: It is a prospective study, 35 laryngectomized patients were sedated with midazolam and underwent digestive endoscopy and tracheoesophageal punch with vocal prosthesis insertion. RESULTS: A success rate of 94.2% was achieved with this surgical technique. The mean procedure time was estimated at 12 minutes, and no serious complications due to the prosthesis insertion were observed. CONCLUSION: The advantages of this new technique over the classic technique are lack of use of general anesthesia, performance of procedure on an outpatient basis, lower complication risks (including hemorrhage, mediastinitis, vertebral fracture, esophageal perforation; and minor oropharyngeal, and esophageal mucosal trauma), and direct visualization of the prosthesis in the esophageal lumen. PMID- 12748563 TI - Changes of esophageal motility after total laryngectomy. AB - OBJECTIVE: Total laryngectomy completely interrupts the continuity of the proximal digestive tract and may lead to derangement in esophageal motility. The purpose of this investigation was to find out how total laryngectomy changes the resting and the maximum contracting pressures of the upper esophageal sphincter muscle and how it affects the coordination of the contraction and the relaxation between the pharynx and the upper esophageal sphincter muscles. If changes in the function of the upper esophageal sphincter muscle should occur, this study will also demonstrate how it affects the motility of the esophagus and the lower esophageal sphincter muscle. METHODS: In an attempt to explain postoperative motility changes, the stationary pull through method of manometric evaluation was used to quantify the alteration in esophageal motility. For the manometric evaluation of the esophagus, a polyethylene catheter with 8 internal tubes was used. The study was performed on a group of 15 patients with total laryngectomy and 15 people without esophageal disease or symptoms as the control group. RESULTS: There was a statistically significant difference between the laryngectomy group and the control group for both the resting and maximum contraction pressures as well as for coordination and relaxation of the upper esophageal sphincter. (P < 0.05) In the laryngectomy group, 3 patients who complained of postoperative dysphasia showed more severe functional changes. The proximal esophageal body pressure and peristaltic waves were significantly decreased in the laryngectomy group. No significant difference between the laryngectomy group and the control group was noted in terms of the lower esophageal resting sphincter pressure and the postdeglution pressure. There also was no significant difference between the two groups in the degree of lower esophageal sphincter coordination and relaxation. CONCLUSION: From these results, it may be concluded that interruption of the cricopharyngeal muscle and pharyngeal plexus after laryngectomy not only may produce local derangement of upper esophageal sphincter function but also may produce abnormalities in peristalsis of the proximal esophageal body. However, the function of lower esophageal sphincter did not show any significant difference between the laryngectomy group and the control group. PMID- 12748564 TI - Oncologic and functional results of near-total laryngectomy. AB - OBJECTIVE: To report the oncologic and functional outcome of patients undergoing near-total laryngectomy. STUDY DESIGN AND SETTING: A retrospective analysis was carried out from 1991 through 1998. RESULTS: Eighty-seven patients underwent near total laryngectomy. The Kaplan-Meier overall survival, cause-specific survival, and relapse-free survival estimates at 5 years were 48.2%, 75.8%, and 72.4%, respectively. Univariate analysis revealed prognosis significance for location, stage, and pathologic cervical lymph node status. Five percent of the patients developed local recurrence, 16% recurrence in cervical lymph nodes, and 10% distant metastasis. Pharyngocutaneous fistula was the most frequent complication (48%). Seventy-seven percent of cases achieved voice preservation. Symptomatic aspiration was noted in 12.3% of cases. CONCLUSION: Near-total laryngectomy can be a successful surgical procedure for selected patients; it does not replace total laryngectomy but reduces its indications. Voice preservation can be achieved in most cases. SIGNIFICANCE: Near-total laryngectomy enhances the speech rehabilitation options for the laryngectomy patient by adding a physiologic, nonprosthetic tissue technique. PMID- 12748565 TI - Clinical associations between tinnitus and chronic pain. AB - OBJECTIVE: We sought to estimate the prevalence and severity of tinnitus in patients with chronic pain. STUDY DESIGN AND SETTING: We conducted a prospective nonrandomized study in which a survey and the Tinnitus Handicap Inventory (THI) were distributed at a tertiary chronic pain clinic. RESULTS: Seventy-two patients participated. 50 women (mean age, 53 years) and 22 men (mean age, 47.5 years); 54.2% reported having tinnitus. There was an even distribution of patients reporting the onset of tinnitus as before versus after the onset of pain. Four patients reported a direct association between tinnitus and pain. The mean THI score was 27 (of 100) (n = 35). Fifteen subjects scored less than 16, indicating no handicap, and 4 scored over 58, indicating a severe handicap. CONCLUSION: The study results suggest a high incidence of tinnitus within this population. There were few strong associations between pain and tinnitus. Tinnitus does not significantly handicap the majority of these patients. SIGNIFICANCE: Tinnitus is a common symptom in the chronic pain population but is not a significant problem for these patients. PMID- 12748566 TI - Minimally invasive laser contraction myringoplasty for tympanic membrane atelectasis. AB - OBJECTIVE: We sought to develop a minimally invasive surgical technique using the CO(2) laser to reduce or eliminate tympanic membrane atelectasis in a select group of patients. STUDY DESIGN: Thirty-seven ears with varying degrees of tympanic membrane atelectasis underwent CO(2) laser myringoplasty with the patients under intravenous sedation in the operating room setting. Atelectasis severity was graded for each patient and documented before and after laser myringoplasty through photodocumentation. Patients were followed for 1 year with comparison tympanic membrane photography. SETTING: The study was conducted in a tertiary care private otology-neurotology practice. RESULTS: Laser myringoplasty significantly reduced retraction pocket severity in most patients. No patients required resection of the retraction pocket or tympanoplasty. The most favorable outcomes were observed in patients with atelectasis addressed early rather than later in its more advanced stages. CONCLUSION: Laser contraction myringoplasty can reduce or eliminate atelectatic areas of the tympanic membrane through immediate contraction and "tightening" of the tympanic membrane tissues. Clinicians should use a standardized tympanic membrane atelectasis grading format. SIGNIFICANCE: A minimally invasive surgical technique for addressing tympanic membrane atelectasis is described, and a tympanic membrane atelectasis grading system is presented based on size, location, and depth of the atelectatic region. PMID- 12748567 TI - Systematic approach to benign paroxysmal positional vertigo in the elderly. AB - OBJECTIVE: We evaluated the effectiveness of a management approach that combines the canalith repositioning maneuver (CRM) and vestibular rehabilitation (VR) in the treatment of benign positional paroxysmal vertigo (BPPV) in elderly persons. STUDY DESIGN AND SETTING: Forty-seven patients (>/=70 years old) with the diagnosis of unilateral posterior semicircular canal BPPV formed the study population. This study has 2 parts. In the first part, patients were randomly assigned to 1 of 2 groups: the CRM and avoidance (no treatment). Patients were evaluated 1 month after the first visit. Those patients not responding to treatment were enrolled in the second part of the study, treated with an individualized combination of CRM and VR, and then reevaluated 3 months later. RESULTS: Statistically significant improvement of vertigo and provoked nystagmus in 64% of patients in the CRM group compared with the no-treatment group. After the addition of VR, 77% of all patients improved. CONCLUSION: A combination of CRM and VR improves BPPV in the elderly. SIGNIFICANCE: These findings suggest that although CRM is more effective than no treatment, VR can be added to improve the results in the treatment of BPPV. PMID- 12748568 TI - Bone wax prevents nystagmus after labyrinthine fenestration in guinea pigs. AB - OBJECTIVE: We examined the effect of 3 methods of closure of labyrinthine fenestrations. STUDY DESIGN AND SETTING: A guinea pig animal model was developed to examine the effect of 3 methods of closure of labyrinthine fenestrations. These methods included bone wax alone, fascia and fibrin glue, and fascia alone. Nystagmus, a reproducible sign of labyrinthine injury, was chosen as an end point for comparing materials commonly used in human ear surgery. RESULTS: Animals repaired with bone wax showed no postoperative nystagmus, whereas animals repaired with other materials or that underwent canal fenestration without repair showed 3 to 5 days of postoperative nystagmus. CONCLUSIONS: Although it is premature to extrapolate our results to human surgery, postoperative nystagmus can be eliminated in guinea pigs by sealing labyrinthine fenestrations with bone wax. SIGNIFICANCE: Given the intimate relationship between the vestibular and cochlear systems, it is possible that prevention of vestibular irritation might be associated with preservation of cochlear function. PMID- 12748569 TI - Rate of nitrous oxide exchange across the middle ear mucosa in monkeys before and after blockage of the mastoid antrum. AB - OBJECTIVES: We tested the hypothesis that mastoid volume buffers the rate of change in middle ear pressure caused by transmucosal, inert gas exchange. STUDY DESIGN: Twelve monkeys were randomly assigned to group 1 or group 2. Right ears of group 1 had sham surgery and of group 2 had obstruction of the mastoid antrum. Before and after surgery, the time constant for transmucosal N(2)O exchange was estimated from N(2)O breathing experiments. The hypothesis predicts that the postoperative time constant measured for right ears of group 2 but not group 1 is greater than that measured before surgery. RESULTS: Mastoid antrum block significantly decreased right middle ear volume but did not affect the time constant for transmucosal N(2)O exchange. CONCLUSION: A mastoid gas-reserve function is not supported by the experimental data. SIGNIFICANCE: These results for monkeys and the theory developed to explain the effect of mastoid volume on transmucosal inert gas exchange suggest that the results for previous experiments in humans interpreted as evidencing a mastoid gas-reserve function are consistent with alternative explanations. PMID- 12748570 TI - The biochemical analysis of tympanosclerotic plaques. AB - OBJECTIVE: We conducted a biochemical analysis of tympanosclerotic (TS) plaques. METHODS: Forty-five samples containing TS plaques were obtained from 45 patients. Qualitative and quantitative biochemical analyses were performed. TS plaques were evaluated macroscopically in respect to color and structure. RESULTS: Phosphate, calcium, ammonium, and cholesterol were detected in 44, 43, 35, and 7 samples, respectively. Mean calcium and phosphate levels and protein levels of samples were 2.5 +/- 2.6 mg/100 mg tissue, 0.16 +/- 0.11 mg, and 3.4 +/- 3.4 mg, respectively. TS plaques were dirty white in 20 samples, brunette in 13, yellowish in 4, white in 7, and brown in 1. The structure of TS plaques was hard in consistency in 23 samples, fibrous in 14 samples, and soft in 8 samples. CONCLUSION: The wider range of values and different mineral contents in our series might be explained by the fact that we had a much larger variety of tissues in various stages of the TS process. The color and the consistency of TS plaques are due to the discrepancy in their content and the amount of mineral, cholesterol, and protein. PMID- 12748571 TI - Ankyloglossia: the adolescent and adult perspective. AB - OBJECTIVES: We sought to characterize examination findings and functional limitations due to ankyloglossia in adolescents and adults and to evaluate frenuloplasty in this group. Study design A prospective study was conducted of 15 individuals with ankyloglossia aged 14 to 68 years. Baseline symptoms were recorded by questionnaire, and tongue mobility measures were compared with that of 20 control subjects. Six subjects were reassessed postfrenuloplasty. RESULTS: Thirteen of 14 patients with uncorrected ankyloglossia (93%) noted symptoms including speech problems (50%) and mechanical limitations (57%), such as difficulty licking the lips. Mean tongue protrusion and elevation at baseline measured 15.5 +/- 6.0 mm and 13.6 +/- 8.0 mm, respectively, for patients and 32.0 +/- 3.9 mm and 30.3 +/- 4.9 mm for control subjects (P < 0.001). Postfrenuloplasty, tongue function improved both subjectively and objectively in 6 of 6 patients, with a mean gain of 9.2 mm for protrusion (P < 0.05) and 13.0 mm for elevation (P < 0.001). CONCLUSION: Symptoms related to ankyloglossia are prevalent in this age group and respond favorably to frenuloplasty. PMID- 12748573 TI - Thoracoscopic management of cervical thoracic duct injuries: an alternative approach. PMID- 12748572 TI - Surgery for prolonged parotid duct obstruction: a case report. PMID- 12748574 TI - Cervicofacial hemangioma and its association with PHACE syndrome. PMID- 12748575 TI - Internal jugular phlebectasia surrounded by mature adipose tissue. PMID- 12748576 TI - Concholith: an unusual case. PMID- 12748577 TI - An unusual sharp foreign body in the esophagus and its removal. PMID- 12748578 TI - Tracheal stomal recurrence of tonsillar cancer. PMID- 12748581 TI - Pediatric cancer survivors: Past history and future challenges. PMID- 12748582 TI - Late effects of treatment for cancer during childhood and adolescence. PMID- 12748583 TI - Longitudinal risk-based health care for adult survivors of childhood cancer. PMID- 12748584 TI - [Molecular and quantitative analyses of Malassezia microflora on the skin of atopic dermatitis patients and genotyping of M. globosa DNA]. AB - To elucidate the role of Malassezia species in atopic dermatitis (AD) requires investigation of the Malassezia microflora on the skin of AD patients. Previously, M. furfur was considered the dominant species in the microflora, however, this microorganism has been reclassified into five species and reanalysis of the microflora based on the current Malassezia taxonomy is therefore needed. Malassezia is more difficult to isolate and culture than other pathogenic yeasts such as Candida and Cryptococcus, making it difficult to elucidate the microflora of AD patients accurately. We developed a PCR-based non culture method that does not require the use of isolation or culture techniques. Of the members of the genus Malassezia, M. globosa colonized the skin of both AD patients and healthy subjects more frequently than other Malassezia species. In addition, we found polymorphisms in the intergenic spacer 1 region of the M. globosar RNA gene. The genotypes of the microorganisms obtained from AD patients were significantly different from those obtained from healthy subjects. We believe that a specific genotype of M. globosa is responsible for exacerbation of AD. PMID- 12748585 TI - [Malassezia and atopic dermatitis]. AB - Although many exacerbating factors for atopic dermatitis (AD) have been discussed, we are focusing on fungus antigen as a pathogenesis for this condition. About half of the patients were sensitized by Candida albicans and/or Malassezia furfur (MF) using IgE. Patients with severe eruption tended to have a higher concentration of specific IgE. IgE to purified antigens such as manganese superoxide dismutase (MnSOD), cyclophilin, and Malf2 from MF was also detected, while the pattern of positive IgE was varied among the patients so that the major allergen could not be determined. Skin testing gave a positive reaction to MF after 24 hours as well as an immediate type reaction; this delayed type reaction was AD specific since a small number of patients with bronchial asthma showed a positive response to MF. Peripheral mononuclear cells co-cultured with crude MF antigen in vitro produced IL-5 in some AD patients. This response was correlated with the severity of facial eruption, indicating that Th2 type response to MF might make these eruptions worse. MF was easily detected from various skin regions,but we were not able to explain why fewer colonies were obtained from a region with dermatitis than from a non-dermatitis region. From these results, we speculate there are patients who have IgE and Th2 cells which respond to MF. The exact mechanism, however, is still obscure as to how normal flora such as MF can react and exacerbate AD. Further investigations should be done to learn more about the relationship between AD and MF. PMID- 12748586 TI - [Atopic dermatitis and Malassezia species: a study of antigenic components of Malassezia species for immunoglobulin E of patients with atopic dermatitis]. AB - Antigenic components extracted by treatment with beta-mercaptoethanol from M. furfur, M. globosa, M. restricta, M. slooffiae, and M. sympodialis were studied for immunoglobulin E antibodies in sera of patients with atopic dermatitis. CBB staining and lectin blots of the extracts showed that each Malassezia species contained species-dependent components at the protein level. In a Western blot with the 2-ME extracts,IgE antibodies against the Malassezia species were found in sera of 83% (for M. globosa), 74% (for M. sympodialis), 65% (for M. furfur), 56% (for M. restricta) and 50% (for M. slooffiae) of the AD patients. In the Western blot inhibition test, the 2-ME extract of M. globosa partially inhibited the reaction of the antigenic components of other Malassezia species with the patients IgE antibodies. These results indicated that Malassezia species contained both species-specific and common antigenic components at the IgE antibody level. A major component of M. globosa was isolated from the 2-ME extract of this fungus by ion-exchange column chromatography and was referred to as Malg46b. Dot blot with the Malg46b containing fraction immunologically reacted with 69% of the sera of the patients, and with 83% of the sera of those who were positive for IgE antibodies to the 2-ME extract of M. globosa in Western blot. The intensities generated for each dot correlated well with the total intensities generated for the 2-ME extract of M. globosa in Western blot (r=0.763) The polyclonal antibody to Malg46b reacted strongly only with the 2-ME extract of M. globosa and reacted slightly with M. restricta. These results indicate that a glycoprotein, Malg46b of M. globosa, is dominantly expressed in this fungus and is a possible major antigen for IgE antibodies in patients with AD. PMID- 12748587 TI - [Seborrheic dermatitis--clinical diagnosis and therapeutic value of different drugs]. AB - Seborrheic dermatitis (SD) is a disease characterized by erythema and accompanied by greasy scale in the seborrheic region. The mechanism by which the disease occurs is still unknown. The genus Malassezia is involved in aggravating SD. Objective diagnosis of SD has yet been established. Atopic dermatitis and psoriasis or contact dermatitis are often confused with SD. One method to differentiate SD from other skin diseases is direct microscopic examination. Mild corticosteroids are effective in treatment of this condition, although, many cases recur within a few days. Antifungal agents are also effective in the treatment of SD by reducing the number of spores, which results in prolongation of the time to recurrence. It is my recommendation that antifungal agents be the first choice of therapy. PMID- 12748588 TI - [Genomic analysis in Candida albicans]. AB - With the recent advances in DNA sequencing technology, a succession of entire genome sequences have been published. A number of genome projects are underway in pathogenic fungi. From these, we present the history and current status of the genomic analysis of Candida albicans. The sequencing project for this organism has been undertaken at Stanford University, and is now nearing the end. PMID- 12748589 TI - [Azole resistance in Candida spp]. AB - The emergence of azole-resistant Candida spp. is a significant problem after long term treatment of recurrent oropharyngeal candidiasis in HIV-infected patients. Several mechanisms can cause this resistance. An important mechanism of azole resistance is reduced intracellular accumulation of the drug. Among the multidrug efflux transporters, ABC transporters and the major facilitator superfamily are reported to cause the resistance. Erg11p, sterol C14 alfa-demethylase, is a target of azole derivatives. It was reported that ERG11 over-expression had only a modest effect on the development of azole resistance. However, mutations in the ERG11 gene can cause the resistance, probably by reducing binding of azole to the target enzyme. We sequenced the ERG11 gene in a high-level azole resistant C. albicans strain, Darlington, and found that two amino acid substitutions, Y132H and I471T, had been encoded in the Darlington ERG11 gene. To assess the significance of these substitutions, we replaced one of the two copies of ERG11 gene in anazole-susceptible strain of C. albicans with a copy of the Darlington ERG11 and this resulted in a modest increase in azole resistance. Furthermore, to estimate the effect of Y132H and I471T individually, ERG11 genes with either or both mutations were expressed in S. cerevisiae. The I471T substitution, not previously described, conferred azole resistance when overexpressed alone and increased this resistance when added to the Y132H substitution. Alterations in the sterol biosynthetic pathway are another resistance mechanism. Inhibition of 14 alfa-demethylase by azole results not only in ergosterol depletion but also in accumulation of methylated sterol 14 alfa-methylergosta-8, 22(28)-dien-3 beta, 6 alfa-diol. We deleted the ERG3 gene, which encodes a sterol 5, 6-desaturase, in C. albicans, and the deletion resulted in reduced susceptibility of the mutant to azoles.Sterol analysis revealed that erg3 mutant lost both ergosterol and diol when cultured with fluconazole. PMID- 12748590 TI - [Subtractive gene cloning and gene-disruption for elucidation of pseudohyphal formation in Candida tropicalis]. AB - The dimorphic transition from yeast to pseudohyphae in the petroleum-assimilating yeast Candida tropicalis occurs following the addition of ethanol to glucose semi defined medium. Subtractive gene cloning was performed on the cDNA from the yeast growing control culture and on that from the ethanol-supplemented one (the ethanol culture). A homologue of Schizosaccharomyces pombe nmt1+ or Saccharomyces cerevisiae THI5 was isolated from the cDNA fraction as a preferentially expressed gene for the ethanol culture. This homologue was tentatively called Ctnmt1+, since exogenous thiamine repressed its expression in C. tropicalis growth media. The ethanol culture showed a biphasic pattern of growth phases and the expression of Ctnmt1+ occurred at the first growth phase. The supplementation of thiamine to the ethanol culture at the first phase was followed by repression of Ctnmt1+ expression and also delay of pseudohyphal growth: filamentous growth was inhibited and chains of yeast cells were formed. A Ctnmt1+ disruptant of this organism did not show thiamine auxotrophy and produced pseudohyphal filaments even in the control culture. The supplementation of oxythiamine, an analog of thiamine, to the control culture was followed by the appearance of pseudohyphal filaments, indicating the participation of thiamine during the process of pseudohyphal growth in this organism. PMID- 12748591 TI - [Isolation and molecular characterization of the CaPHO85 gene: a negative regulator of phosphate metabolism (PHO system) in Candida albicans]. AB - The PHO system is an ingenious mechanism by which the yeast Saccharomyces cerevisiae regulates the expression of a set of genes involved in phosphate metabolism in response to the change of phosphate concentrations in the environment. A key factor in this mechanism is the Pho85 kinase, which has been discovered as a negative regulator of the PHO system. One of the genes isolated in our laboratory in screening the protein kinase genes from Candida albicans was identified as a homologue (CaPHO85) of the PHO85 of S. cerevisiae, based on the following results. a) Pho85 is the polypeptide with the highest homology to CaPho85 (62% identity) among the S. cerevisiae genome sequence. b) The position of insertion of the intron is quite similar between CaPHO85 (in the 7th codon of the N-terminal MTGSSSQ) and S. cerevisiae PHO85 (in the 6th codon of the N terminal MSSSSQ). c) The nucleotide sequences in the intron possess the consensus sequences for yeast intron: the 5-splice-site, internal, and the 3-splice-site sequence. d) CaPHO85 complemented the S. cerevisiae pho85 mutation. e) CaPho85 contains all of the consensus sequences for the ATP-binding domain and for the kinase domain found in S. cerevisiae Pho85. PMID- 12748592 TI - [Evaluation of targets for antifungal drugs using a system to regulate gene expression]. AB - A system to regulate gene expression is a convenient tool to explore gene function(s) not only in prokaryote but also in eukaryote. Such manipulation tools are scarce in the medical mycology field due to its complexity and diploidity. Although systems to regulate gene expression have been constructed, most of them are restricted in their application to particular culture conditions due to the nature of the promoter used. This motivated us to establish a new regulatable expression system that can function regardless of culture conditions, including in a host. In this review, a new system using tetracycline or its derivative as a molecular switch is introduced, which can function in several culture conditions, and in a host. We also show that the system can be applied to the selection of antifungal drug targets, which is the first step in a target-based strategy for drug discovery. PMID- 12748593 TI - [Guidelines for clinical evaluation of topical antifungal agents]. AB - The Japanese Society for Medical Mycology (JSMM) decided in 2002 to establish guidelines for the clinical evaluation of antifungal agents. The JSMM committee presents here guidelines for the clinical evaluation of topical antifungal agents in the dermatology field. Guidelines for the Clinical Evaluation of Antibiotic Agents established by the Japanese Society of Chemotherapy were referred to, and the diseases subjected to clinical evaluation include tinea (tinea pedis and tinea glabrosa), cutaneous candidiasis, and pityriasis versicolor. Among superficial mycoses, tinea pedis is viewed as the pivotal disease because it is intractable and is the most common. Therefore, the clinical efficacy of antifungal agents for external use in this condition should be established, and tinea pedis is subjected to phase III clinical studies. If efficacy of the antifungal agents is confirmed in the treatment of tinea pedis, a comparative study need not necessarily be performed for tinea glabrosa. If the number of patients is adequate for statistical analysis, a comparative study should be considered for both cutaneous candidiasis and pityriasis versicolor. However, if the number of patients is low, the efficacy of the agents should be evaluated based on their antifungal activity on pathogens and the results of open trials, and a comparative study is not necessarily performed for such diseases. The safety should be strictly evaluated. PMID- 12748594 TI - [A short-term treatment of tinea corporis and tinea cruris with oral terbinafine]. AB - We studied the effectiveness of short-term treatment of tinea corporis and tinea cruris with oral terbinafine at 250 mg/day for 2-3 days. The treatment on an open study basis consisted of two groups: the first group (n=17) was given 250 mg/day for two consecutive days, and the second group (n=24) was given the same dose for three consecutive days. No patient was treated topically. Effectiveness was evaluated at the end of the second week both clinically and mycologically (KOH examination and culture). In the two-day group, five cases showed an excellent response, three had a good response and nine had a fairly good response. Patients with good response or better comprised 47.1% of the total, while those with excellent response stood at 29.4%. The negative mycological examination ratio was 47.1%. In the three-day group, 12 cases showed an excellent response, four a good response while eight had a fairly good response. Patients with a good response or better comprised 66.7% of the group, while cases with an excellent response comprised 50.0%. The negative mycological examination ratio stood at 66.7%. The overall effectiveness evaluation showed no statistically significant difference between the two treatment groups in the Wilcoxon's rank sum test. No side effect was observed in either group. These findings showed that terbinafine therapy of tinea cruris is effective even with a short-term treatment of 2-3 days at a small dose. PMID- 12748596 TI - The role of chlamydospores of Paracoccidioides brasiliensis. AB - The role of chlamydospores in the conversion process from a mycelial-to-yeast form using the slide culture method was studied. Three clinical isolates and two other isolates from armadillo, belonging to the fungal species Paracoccidioides brasiliensis, were cultured on Sabouraud dextrose agar (SDA), potato dextrose agar (PDA) and brain heart infusion dextrose agar (BHIDA). Initially, the mycelial forms of each isolate were grown at 25 degrees C for 7, 14, 30 or 60 days on slide cultures and then the temperature was shifted to 35 degrees C. Interestingly, the slide cultures of all the isolates at 25 degrees C formed chlamydospores on either SDA or BHIDA, whereas, on PDA medium, aleurioconidia were formed. If the slide cultures on BHIDA were incubated at 35 degrees C for 7 to 14 days, multiple budding forms could be observed. This phenomenon was not evident in the slide cultures of SDA or PDA. The results of this morphological study indicate that in P. brasiliensis, chlamydospores may play an important role in the conversion process from a mycelial-to-yeast form. PMID- 12748595 TI - [Prophylactic efficacy of a basidiomycetes preparation AHCC against lethal Candida albicans infection in experimental granulocytopenic mice]. AB - The prophylactic effects of a Basidiomycetes preparation, AHCC, against lethal Candida albicans infection were investigated in non treated or immunosuppressed mice. In the cyclophosphamide-or 5-fluorouracil (5-FU)-treated leukopenic mice and nontreated mice, the intraperitoneal administration of AHCC prior to C. albicans infection clearly prolonged the survival periods of the infected mice. In doxorubicin-treated mice, AHCC was less but significantly effective. On the other hand, in prednisolone-treated mice, AHCC was not effective. Oral administration of AHCC also protected the 5-FU-treated mice from lethal Candida infection, as indicated by prolongation of the survival periods and inhibition of Candida growth in the kidneys of these mice and by the increase in a number of neutrophils in their peripheral blood. These results suggested that AHCC may display a protective role against opportunistic fungal infection in leukopenic hosts. PMID- 12748598 TI - Don't mention the war. PMID- 12748597 TI - Immunomagnetic isolation of Cryptococcus neoformans by beads coated with anti Cryptococcus serum. AB - Immunomagnetic separation (IMS) was utilized for the selective isolation of Cryptococcus neoformans from environmental sources, such as soils and pigeon droppings. Magnetic beads coated with anti-cryptococcal IgG (serotypes A and B) were used to isolate the fungus. In a modeled spiking experiment using C. neoformans serotype A strain and anti-serotype A antibody, the recovery rate of the cells was more than 47%. Specificity experiments using C. neoformans and Candida albicans showed that the beads, when coated with specific antibody for C. neoformans, were highly effective for the separation of C. neoformans strains from C. albicans (more than 97%). The IMS of serotype B cells with purified anti serotype B antibody indicated a high specificity. When this IMS technique was applied to soils and pigeon droppings, C. neoformans cells were selectively isolated from 3 out of 8 samples, and C. neoformans DNAs were identified by PCR. Therefore C. neoformans cells were thus selectively isolated and the efficiency of the technique further confirmed. PMID- 12748599 TI - Gene patents and the public good. PMID- 12748601 TI - Researchers rue hasty destruction of wheat. PMID- 12748600 TI - Biologists take tentative steps towards bespoke cancer drugs. PMID- 12748602 TI - Cash crisis casts pall over synchrotron X-ray source. PMID- 12748603 TI - US support for Spain triggers unease over fusion project. PMID- 12748605 TI - Asteroid probe to test technologies. PMID- 12748604 TI - Medical council pins hopes on public advocate of science. PMID- 12748606 TI - Europe dithers as Canada cuts cod fishing. PMID- 12748607 TI - Low embryo count fuels US stem-cell debate. PMID- 12748608 TI - MIT pulls out of Asian Media Lab in argument over role. PMID- 12748610 TI - The man they love to hate. PMID- 12748611 TI - Conservation biology: Mock turtles. PMID- 12748612 TI - Making a song and dance about emotion. PMID- 12748613 TI - Early work on elephant gait not to be forgotten. PMID- 12748614 TI - Japanese system buries the individual researcher. PMID- 12748620 TI - Mental self: The person within. PMID- 12748621 TI - Plant reproduction: Sex and self-denial. PMID- 12748622 TI - Cosmology: A just-so story. PMID- 12748623 TI - Stem cells: Self-renewal writ in blood. PMID- 12748625 TI - Genomics: Yeast rises again. PMID- 12748627 TI - Planetary science: Jupiter's moonopoly. PMID- 12748626 TI - Molecular biology: Disruptive influence. PMID- 12748631 TI - Planetary science: Mission to Earth's core--a modest proposal. PMID- 12748629 TI - Obituary: Charles A. Janeway Jr (1943-2003). PMID- 12748632 TI - Aetiology: Koch's postulates fulfilled for SARS virus. PMID- 12748633 TI - Sequencing and comparison of yeast species to identify genes and regulatory elements. AB - Identifying the functional elements encoded in a genome is one of the principal challenges in modern biology. Comparative genomics should offer a powerful, general approach. Here, we present a comparative analysis of the yeast Saccharomyces cerevisiae based on high-quality draft sequences of three related species (S. paradoxus, S. mikatae and S. bayanus). We first aligned the genomes and characterized their evolution, defining the regions and mechanisms of change. We then developed methods for direct identification of genes and regulatory motifs. The gene analysis yielded a major revision to the yeast gene catalogue, affecting approximately 15% of all genes and reducing the total count by about 500 genes. The motif analysis automatically identified 72 genome-wide elements, including most known regulatory motifs and numerous new motifs. We inferred a putative function for most of these motifs, and provided insights into their combinatorial interactions. The results have implications for genome analysis of diverse organisms, including the human. PMID- 12748634 TI - An abundant population of small irregular satellites around Jupiter. AB - Irregular satellites have eccentric orbits that can be highly inclined or even retrograde relative to the equatorial planes of their planets. These objects cannot have formed by circumplanetary accretion, unlike the regular satellites that follow uninclined, nearly circular and prograde orbits. Rather, they are probably products of early capture from heliocentric orbits. Although the capture mechanism remains uncertain, the study of irregular satellites provides a window on processes operating in the young Solar System. Families of irregular satellites recently have been discovered around Saturn (thirteen members, refs 6, 7), Uranus (six, ref. 8) and Neptune (three, ref. 9). Because Jupiter is closer than the other giant planets, searches for smaller and fainter irregular satellites can be made. Here we report the discovery of 23 new irregular satellites of Jupiter, so increasing the total known population to 32. There are five distinct satellite groups, each dominated by one relatively large body. The groups were most probably produced by collisional shattering of precursor objects after capture by Jupiter. PMID- 12748635 TI - Chaos-assisted capture of irregular moons. AB - It has been thought that the capture of irregular moons--with non-circular orbits -by giant planets occurs by a process in which they are first temporarily trapped by gravity inside the planet's Hill sphere (the region where planetary gravity dominates over solar tides). The capture of the moons is then made permanent by dissipative energy loss (for example, gas drag) or planetary growth. But the observed distributions of orbital inclinations, which now include numerous newly discovered moons, cannot be explained using current models. Here we show that irregular satellites are captured in a thin spatial region where orbits are chaotic, and that the resulting orbit is either prograde or retrograde depending on the initial energy. Dissipation then switches these long-lived chaotic orbits into nearby regular (non-chaotic) zones from which escape is impossible. The chaotic layer therefore dictates the final inclinations of the captured moons. We confirm this with three-dimensional Monte Carlo simulations that include nebular drag, and find good agreement with the observed inclination distributions of irregular moons at Jupiter and Saturn. In particular, Saturn has more prograde irregular moons than Jupiter, which we can explain as a result of the chaotic prograde progenitors being more efficiently swept away from Jupiter by its galilean moons. PMID- 12748636 TI - A theory of power-law distributions in financial market fluctuations. AB - Insights into the dynamics of a complex system are often gained by focusing on large fluctuations. For the financial system, huge databases now exist that facilitate the analysis of large fluctuations and the characterization of their statistical behaviour. Power laws appear to describe histograms of relevant financial fluctuations, such as fluctuations in stock price, trading volume and the number of trades. Surprisingly, the exponents that characterize these power laws are similar for different types and sizes of markets, for different market trends and even for different countries--suggesting that a generic theoretical basis may underlie these phenomena. Here we propose a model, based on a plausible set of assumptions, which provides an explanation for these empirical power laws. Our model is based on the hypothesis that large movements in stock market activity arise from the trades of large participants. Starting from an empirical characterization of the size distribution of those large market participants (mutual funds), we show that the power laws observed in financial data arise when the trading behaviour is performed in an optimal way. Our model additionally explains certain striking empirical regularities that describe the relationship between large fluctuations in prices, trading volume and the number of trades. PMID- 12748637 TI - Measurement of the displacement field of dislocations to 0.03 A by electron microscopy. AB - Defects and their associated long-range strain fields are of considerable importance in many areas of materials science. For example, a major challenge facing the semiconductor industry is to understand the influence of defects on device operation, a task made difficult by the fact that their interactions with charge carriers can occur far from defect cores, where the influence of the defect is subtle and difficult to quantify. The accurate measurement of strain around defects would therefore allow more detailed understanding of how strain fields affect small structures-in particular their electronic, mechanical and chemical properties--and how such fields are modified when confined to nanometre sized volumes. Here we report the measurement of displacements around an edge dislocation in silicon using a combination of high-resolution electron microscopy and image analysis inherited from optical interferometry. The agreement of our observations with anisotropic elastic theory calculations is better than 0.03 A. Indeed, the results can be considered as an experimental verification of anisotropic theory at the near-atomic scale. With the development of nanostructured materials and devices, we expect the use of electron microscopy as a metrological tool for strain analysis to become of increasing importance. PMID- 12748638 TI - Detection of bromine monoxide in a volcanic plume. AB - The emission of volcanic gases usually precedes eruptive activity, providing both a warning signal and an indication of the nature of the lava soon to be erupted. Additionally, volcanic emissions are a significant source of gases and particles to the atmosphere, influencing tropospheric and stratospheric trace-gas budgets. Despite some halogen species having been measured in volcanic plumes (mainly HCl and HF), little is known about bromine compounds and, in particular, gas-phase reactive bromine species. Such species are especially important in the stratosphere, as reactive bromine-despite being two orders of magnitude less abundant than chlorine-accounts for about one-third of halogen-catalysed ozone depletion. In the troposphere, bromine-catalysed complete ozone destruction has been observed to occur regularly during spring in the polar boundary layers as well as in the troposphere above the Dead Sea basin. Here we report observations of BrO and SO2 abundances in the plume of the Soufriere Hills volcano (Montserrat) in May 2002 by ground-based multi-axis differential optical absorption spectroscopy. Our estimate of BrO emission leads us to conclude that local ozone depletion and small ozone 'holes' may occur in the vicinity of active volcanoes, and that the amount of bromine emitted from volcanoes might be sufficiently large to play a role not only in the stratosphere, but also in tropospheric chemistry. PMID- 12748639 TI - Parallel extinction risk and global distribution of languages and species. AB - There are global threats to biodiversity with current extinction rates well above background levels. Although less well publicized, numerous human languages have also become extinct, and others are threatened with extinction. However, estimates of the number of threatened languages vary considerably owing to the wide range of criteria used. For example, languages have been classified as threatened if the number of speakers is less than 100, 500, 1,000, 10,000, 20,000 or 100,000 (ref. 3). Here I show, by applying internationally agreed criteria for classifying species extinction risk, that languages are more threatened than birds or mammals. Rare languages are more likely to show evidence of decline than commoner ones. Areas with high language diversity also have high bird and mammal diversity and all three show similar relationships to area, latitude, area of forest and, for languages and birds, maximum altitude. The time of human settlement has little effect on current language diversity. Although similar factors explain the diversity of languages and biodiversity, the factors explaining extinction risk for birds and mammals (high altitude, high human densities and insularity) do not explain the numbers of endangered languages. PMID- 12748640 TI - Rapid worldwide depletion of predatory fish communities. AB - Serious concerns have been raised about the ecological effects of industrialized fishing, spurring a United Nations resolution on restoring fisheries and marine ecosystems to healthy levels. However, a prerequisite for restoration is a general understanding of the composition and abundance of unexploited fish communities, relative to contemporary ones. We constructed trajectories of community biomass and composition of large predatory fishes in four continental shelf and nine oceanic systems, using all available data from the beginning of exploitation. Industrialized fisheries typically reduced community biomass by 80% within 15 years of exploitation. Compensatory increases in fast-growing species were observed, but often reversed within a decade. Using a meta-analytic approach, we estimate that large predatory fish biomass today is only about 10% of pre-industrial levels. We conclude that declines of large predators in coastal regions have extended throughout the global ocean, with potentially serious consequences for ecosystems. Our analysis suggests that management based on recent data alone may be misleading, and provides minimum estimates for unexploited communities, which could serve as the 'missing baseline' needed for future restoration efforts. PMID- 12748641 TI - Attractor dynamics of network UP states in the neocortex. AB - The cerebral cortex receives input from lower brain regions, and its function is traditionally considered to be processing that input through successive stages to reach an appropriate output. However, the cortical circuit contains many interconnections, including those feeding back from higher centres, and is continuously active even in the absence of sensory inputs. Such spontaneous firing has a structure that reflects the coordinated activity of specific groups of neurons. Moreover, the membrane potential of cortical neurons fluctuates spontaneously between a resting (DOWN) and a depolarized (UP) state, which may also be coordinated. The elevated firing rate in the UP state follows sensory stimulation and provides a substrate for persistent activity, a network state that might mediate working memory. Using two-photon calcium imaging, we reconstructed the dynamics of spontaneous activity of up to 1,400 neurons in slices of mouse visual cortex. Here we report the occurrence of synchronized UP state transitions ('cortical flashes') that occur in spatially organized ensembles involving small numbers of neurons. Because of their stereotyped spatiotemporal dynamics, we conclude that network UP states are circuit attractors--emergent features of feedback neural networks that could implement memory states or solutions to computational problems. PMID- 12748642 TI - Turning on and off recurrent balanced cortical activity. AB - The vast majority of synaptic connections onto neurons in the cerebral cortex arise from other cortical neurons, both excitatory and inhibitory, forming local and distant 'recurrent' networks. Although this is a basic theme of cortical organization, its study has been limited largely to theoretical investigations, which predict that local recurrent networks show a proportionality or balance between recurrent excitation and inhibition, allowing the generation of stable periods of activity. This recurrent activity might underlie such diverse operations as short-term memory, the modulation of neuronal excitability with attention, and the generation of spontaneous activity during sleep. Here we show that local cortical circuits do indeed operate through a proportional balance of excitation and inhibition generated through local recurrent connections, and that the operation of such circuits can generate self-sustaining activity that can be turned on and off by synaptic inputs. These results confirm the long-hypothesized role of recurrent activity as a basic operation of the cerebral cortex. PMID- 12748643 TI - A progeroid syndrome in mice is caused by defects in A-type lamins. AB - Numerous studies of the underlying causes of ageing have been attempted by examining diseases associated with premature ageing, such as Werner's syndrome and Hutchinson-Gilford progeria syndrome (HGPS). HGPS is a rare genetic disorder resulting in phenotypes suggestive of accelerated ageing, including shortened stature, craniofacial disproportion, very thin skin, alopecia and osteoporosis, with death in the early teens predominantly due to atherosclerosis. However, recent reports suggest that developmental abnormalities may also be important in HGPS. Here we describe the derivation of mice carrying an autosomal recessive mutation in the lamin A gene (Lmna) encoding A-type lamins, major components of the nuclear lamina. Homozygous mice display defects consistent with HGPS, including a marked reduction in growth rate and death by 4 weeks of age. Pathologies in bone, muscle and skin are also consistent with progeria. The Lmna mutation resulted in nuclear morphology defects and decreased lifespan of homozygous fibroblasts, suggesting premature cell death. Here we present a mouse model for progeria that may elucidate mechanisms of ageing and development in certain tissue types, especially those developing from the mesenchymal cell lineage. PMID- 12748644 TI - DNA helicase Srs2 disrupts the Rad51 presynaptic filament. AB - Mutations in the Saccharomyces cerevisiae gene SRS2 result in the yeast's sensitivity to genotoxic agents, failure to recover or adapt from DNA damage checkpoint-mediated cell cycle arrest, slow growth, chromosome loss, and hyper recombination. Furthermore, double mutant strains, with mutations in DNA helicase genes SRS2 and SGS1, show low viability that can be overcome by inactivating recombination, implying that untimely recombination is the cause of growth impairment. Here we clarify the role of SRS2 in recombination modulation by purifying its encoded product and examining its interactions with the Rad51 recombinase. Srs2 has a robust ATPase activity that is dependent on single stranded DNA (ssDNA) and binds Rad51, but the addition of a catalytic quantity of Srs2 to Rad51-mediated recombination reactions causes severe inhibition of these reactions. We show that Srs2 acts by dislodging Rad51 from ssDNA. Thus, the attenuation of recombination efficiency by Srs2 stems primarily from its ability to dismantle the Rad51 presynaptic filament efficiently. Our findings have implications for the basis of Bloom's and Werner's syndromes, which are caused by mutations in DNA helicases and are characterized by increased frequencies of recombination and a predisposition to cancers and accelerated ageing. PMID- 12748645 TI - The Srs2 helicase prevents recombination by disrupting Rad51 nucleoprotein filaments. AB - Homologous recombination is a ubiquitous process with key functions in meiotic and vegetative cells for the repair of DNA breaks. It is initiated by the formation of single-stranded DNA on which recombination proteins bind to form a nucleoprotein filament that is active in searching for homology, in the formation of joint molecules and in the exchange of DNA strands. This process contributes to genome stability but it is also potentially dangerous to cells if intermediates are formed that cannot be processed normally and thus are toxic or generate genomic rearrangements. Cells must therefore have developed strategies to survey recombination and to prevent the occurrence of such deleterious events. In Saccharomyces cerevisiae, genetic data have shown that the Srs2 helicase negatively modulates recombination, and later experiments suggested that it reverses intermediate recombination structures. Here we show that DNA strand exchange mediated in vitro by Rad51 is inhibited by Srs2, and that Srs2 disrupts Rad51 filaments formed on single-stranded DNA. These data provide an explanation for the anti-recombinogenic role of Srs2 in vivo and highlight a previously unknown mechanism for recombination control. PMID- 12748648 TI - The complexities of skeletal biology. AB - For a long time, the skeleton was seen as an amorphous tissue of little biological interest. But such a view ignored the large number of genetic and degenerative diseases affecting this organ. Over the past 15 years, molecular and genetic studies have modified our understanding of skeletal biology. By so doing this progress has affected our understanding of diseases and suggested in many instances new therapeutic opportunities. PMID- 12748649 TI - Deciphering skeletal patterning: clues from the limb. AB - Even young children can distinguish a Tyrannosaurus rex from a Brontosaurus by observing differences in bone size, shape, number and arrangement, that is, skeletal pattern. But despite our extensive knowledge about cartilage and bone formation per se, it is still largely a mystery how skeletal pattern is established. Much of what we do know has been learned from studying limb development in chicken and mouse embryos. Based on the data from such studies, models for how limb skeletal pattern is established have been proposed and continue to be hotly debated. PMID- 12748650 TI - Cranial skeletal biology. AB - To artists, the face is a mirror of the soul. To biologists, the face reflects remarkable structural diversity--think of bulldogs and wolfhounds or galapagos finches. How do such variations in skeletal form arise? Do the same mechanisms control skeletogenesis elsewhere in the body? The answers lie in the molecular machinery that generates neural crest cells, controls their migration, and guides their differentiation to cartilage and bone. PMID- 12748651 TI - Developmental regulation of the growth plate. AB - Vertebrates do not look like jellyfish because the bones of their skeletons are levers that allow movement and protect vital organs. Bones come in an enormous variety of shapes and sizes to accomplish these goals, but, with few exceptions, use one process--endochondral bone formation--to generate the skeleton. The past few years have seen an enormous increase in understanding of the signalling pathways and the transcription factors that control endochondral bone development. PMID- 12748652 TI - Osteoclast differentiation and activation. AB - Osteoclasts are specialized cells derived from the monocyte/macrophage haematopoietic lineage that develop and adhere to bone matrix, then secrete acid and lytic enzymes that degrade it in a specialized, extracellular compartment. Discovery of the RANK signalling pathway in the osteoclast has provided insight into the mechanisms of osteoclastogenesis and activation of bone resorption, and how hormonal signals impact bone structure and mass. Further study of this pathway is providing the molecular basis for developing therapeutics to treat osteoporosis and other diseases of bone loss. PMID- 12748653 TI - The genetic basis for skeletal diseases. AB - We walk, run, work and play, paying little attention to our bones, their joints and their muscle connections, because the system works. Evolution has refined robust genetic mechanisms for skeletal development and growth that are able to direct the formation of a complex, yet wonderfully adaptable organ system. How is it done? Recent studies of rare genetic diseases have identified many of the critical transcription factors and signalling pathways specifying the normal development of bones, confirming the wisdom of William Harvey when he said: "nature is nowhere accustomed more openly to display her secret mysteries than in cases where she shows traces of her workings apart from the beaten path". PMID- 12748654 TI - Control of osteoblast function and regulation of bone mass. AB - The skeleton is an efficient 'servo' (feedback-controlled/steady-state) system that continuously integrates signals and responses which sustain its functions of delivering calcium while maintaining strength. In many individuals, bone mass homeostasis starts failing in midlife, leading to bone loss, osteoporosis and debilitating fractures. Recent advances, spearheaded by genetic information, offer the opportunity to stop or reverse this downhill course. PMID- 12748655 TI - Evolving concepts of rheumatoid arthritis. AB - Rheumatoid arthritis is the most common inflammatory arthritis and is a major cause of disability. It existed in early Native American populations several thousand years ago but might not have appeared in Europe until the 17th century. Early theories on the pathogenesis of rheumatoid arthritis focused on autoantibodies and immune complexes. T-cell-mediated antigen-specific responses, T-cell-independent cytokine networks, and aggressive tumour-like behaviour of rheumatoid synovium have also been implicated. More recently, the contribution of autoantibodies has returned to the forefront. Based on the pathogenic mechanisms, specific therapeutic interventions can be designed to suppress synovial inflammation and joint destruction in rheumatoid arthritis. PMID- 12748656 TI - Biotech's balancing act. PMID- 12748657 TI - Language lessons: Heidelberg. PMID- 12748658 TI - HLA-identical stem cell transplantation: is there an optimal CD34 cell dose? AB - A review of the published literature, supplemented with a recent analysis of Fred Hutchinson data, has been undertaken to investigate the association of infused CD34 cell dose with various clinical outcomes after HLA-identical transplantation. Separate assessments for unrelated vs related donors and the use of bone marrow or mobilized G-PBMC have been incorporated. The three primary findings are: (1) higher CD34 dose results in better neutrophil and platelet recovery in all settings; (2) high CD34 doses (>8 x 10(6)/kg) are associated with the development of more chronic GVHD when using related G-PBMC; (3) higher CD34 dose is correlated with improved survival after bone marrow transplantation, especially with unrelated donors. This is not seen when using G-PBMC. The data suggest that the CD34 content of the graft can have a significant impact on clinical outcome after allogeneic transplantation, but optimal dose is dependent on both donor type and stem cell source. PMID- 12748659 TI - Low-intensity hematopoietic stem-cell transplantation across human leucocyte antigen barriers in dyskeratosis congenita. AB - Since the results of conventional hematopoietic stem-cell transplantation (HSCT) for patients with dyskeratosis congenita (DC) are poor owing to the high incidence of transplant-related complications, we explored the use of a low intensity HSCT regimen. We report two children with DC with severe cytopenia, who underwent successful HSCT from a matched unrelated donor after conditioning with fludarabine, cyclophosphamide, and antithymocyte globulin. Graft-versus-host disease (GVHD) prophylaxis consisted of corticosteroids and cyclosporin A. The regimen was well tolerated, no significant transplant-related complications were observed, and engraftment was rapid and complete. At 15 and 16 months after HSCT, the children were fully engrafted, in excellent clinical condition, full-donor chimerism, and no signs of GVHD. We conclude that a low-intensity regimen is sufficient to induce durable engraftment using matched unrelated donor HSCT in DC patients, with minimal 1-year transplant-related toxicity. Longer follow-up will determine whether this regimen also reduces long-term toxicity. PMID- 12748660 TI - Influence of preapheresis clinical factors on the efficiency of CD34+ cell collection by large-volume apheresis. AB - We evaluated 120 leukapheresis procedures (93 patients), in order to detect clinical factors that influence the efficiency of CD34+ collection using Cobe Spectra trade mark cell separators. Hematocrit was >27% and platelet count >30 000/microl in >95% of patients. Platelet transfusions were given if the postprocedure count was &<20 000/microl. Multiple regression analysis was used to analyze putative factors, and a predictive equation defined by stepwise regression modeling. The mean efficiency was 0.59 (s.d. 0.27). Sex (M>F; P=0.01), the volume processed (inversely; P=0.01) and CD34+ cell count (inversely; P=0.04) were associated with efficiency, whereas hematocrit, platelet or leukocyte count, catheter type and patient weight were not. The effect size for predictive factors was small (R(2)=0.21). Adverse events were limited to hypocalcemia. We conclude that female sex, volume processed and CD34+ cell count adversely influence the efficiency of CD34+ cell leukapheresis. However, the impact of volume and CD34+ cell count is small, and likely to be offset by the influence of these same factors on overall yield. Leukapheresis appears to be safe and efficient for autologous blood and marrow transplantation patients with hematocrit >27% and platelet count >30 000/microl. PMID- 12748662 TI - Ideal or actual body weight to calculate CD34+ cell doses for autologous hematopoietic stem cell transplantation? AB - The number of CD34+ cells infused influences the speed of hematologic recovery post-transplant. There are limited data on whether ideal (IBW) or actual (ABW) body weight should be used to calculate CD34+ cell dose. We compared the correlation between recovery to 0.5 x 10(9)/l neutrophils and the CD34+ cell dose based upon ABW as well as IBW in 87 patients autografted for cancer. ABW was >or=25% over IBW in 43% of patients. The median number of CD34+ cells administered was 3.6 x 10(6)/kg ABW and 4.2 x 10(6)/kg IBW. The time to neutrophil recovery was 8-15 days (median 10). There was a stronger inverse correlation between CD34+ cell dose/IBW and neutrophil recovery (r(2)=0.308; P<0.0001) than between CD34+ cell dose/ABW and neutrophil recovery (r(2)=0.267; P<0.0001). The median time to neutrophil recovery was comparable for those receiving >or=2 x 10(6)/kg CD34+ cells/kg IBW as well as ABW (10 days) and those receiving >or=2 x 10(6)/kg CD34+ cells/kg IBW but <2/kg ABW (10 days), but was significantly slower for those receiving <2 x 10(6)/kg CD34+ cells/kg IBW (12 days). These data show that the CD34+ cell dose based on IBW is a better predictor of neutrophil recovery after autotransplantation. PMID- 12748661 TI - Allogeneic bone marrow stem cell transplantation following CD34+ immunomagnetic enrichment in patients with inherited metabolic storage diseases. AB - T-cell depletion is an essential step in reducing the risk of graft-versus-host disease (GVHD) in patients with inherited metabolic storage diseases (IMSD) undergoing hematopoietic stem cell transplantation. This goal can be achieved either by selective removal of T cells or by positive selection of CD34+ cells. Large-scale preparations of purified CD34+ cells from bone marrow products have not been extensively described. We report our results with bone marrow CD34+ cell enrichment using the CliniMACS system in eight children with IMSD. The median recovery of positively selected CD34+ cells was 46.2% with a purity of 97.5%, and a residual T cell content of 0.04 x 10(6). A median of 5.5 x 10(6)/kg of CD34+ cells was infused. All patients engrafted at a median time of 12 days and none of the patients developed GVHD. This method is technically feasible and can be successfully used to transplant children with IMSD. PMID- 12748663 TI - Analysis of immunodominance among minor histocompatibility antigens in allogeneic hematopoietic stem cell transplantation. AB - In major histocompatibility complex (MHC)-matched allogeneic hematopoietic stem cell transplantation (HSCT), donor responses are directed against multiple host minor histocompatibility antigens (mHAgs), producing graft-versus-host disease (GVHD) and graft-versus-tumor (GVT) effects. We studied MHC-matched, mHAg mismatched C3H.SW>C57BL/6 HSCT in which three mHAg are molecularly defined (B6dom1, H3, H13) to determine if there is a hierarchy of immunodominance among the mHAgs and to learn the contribution of each to GVHD. We found that B6dom1 was the immunodominant mHAg. B6dom1 did not block responses to the subdominant mHAgs H3 and H13. The mechanism of immunodominance was not mHAg avidity or affinity for class I. B6dom1 elicited a broader variety of Vbeta clonotypes than either H3 or H13. Severe GVHD could occur in the absence of a strong B6dom1 response. Alloreactivity to isolated B6dom1, H3 or H13 differences did not produce severe GVHD. We concluded that immunodominance is explained by both mHAg density on host cells and the repertoire of donor T cells capable of responding to the mHAgs. Clinically significant GVHD requires donor responses to multiple mHAgs. Modulation of responses to a single immunodominant mHAg is insufficient for the prevention of GVHD, while immunotherapies directed against isolated mHAgs may not provoke severe GVHD. PMID- 12748664 TI - Allogeneic bone marrow transplantation for acute myeloblastic leukaemia in remission: risk factors for long-term morbidity and mortality. AB - In this single-centre retrospective study, we analysed risk factors for nonrelapse long-term morbidity and mortality in patients with acute myeloblastic leukaemia (AML) who had undergone allogeneic transplantation. A total of 112 patients with de novo AML in first complete remission (CR1), n=90 or second complete remission (CR2, n=22) who received un-manipulated bone marrow grafts from human leukocyte antigen identical siblings between January 1985 and August 2000 were included. Of these, 97 patients alive and disease-free for at least 100 days after transplant were selected for the purpose of this long-term analysis. The use of an intensified conditioning regimen, Gram-negative bacteriaemia before transplantation, year of transplantation and number of pretransplant chemotherapy courses for patients in CR1 significantly affected the 7-year event-free survival which was 57%. 7-year transplant-related mortality TRM was 22%. Significant predictors for TRM were: bacterial infections before transplantation, major ABO blood group incompatibility, late severe bacterial infections, and chronic (graft versus-host disease) GvHD. Predictive factors for late severe bacterial infections were infections before transplant, total body irradiation and GvHD. Incidence and risk factors for other late events including, chronic GvHD, late infections, osteonecrosis, cataract, endocrine- cardiac- and lung-complications, cancer and performance status at last follow-up were also studied. The analysis strongly suggests that the combination of pretransplant factors such as chemotherapy and conditioning, and posttransplant factors such as chronic GvHD had a major impact on late nonrelapse morbidity and mortality. PMID- 12748665 TI - Haploidentical vs autologous hematopoietic stem cell transplantation in patients with acute leukemia beyond first remission. AB - This is a retrospective comparison of partially mismatched related donor transplantation (PMRDT) and autotransplantation (ABMT) in advanced acute leukemia. Patients underwent T-cell-depleted PMRDT (n=164) or ABMT (n=131) for acute myeloid leukemia (n=130) or acute lymphoblastic leukemia (n=165). Fewer PMRDT patients were in remission (29 vs 85%; P<0.0001). The 5-year cumulative incidence of transplant-related mortality (TRM) was 52% after PMRDT and 16% after ABMT (P<0.0001). The 5-year cumulative incidence of relapse was 32% after PMRDT and 54% after ABMT (P=0.006). The actuarial unadjusted 5-year disease-free survival (DFS) was 16% after PMRDT and 30% after ABMT. In Cox's regression analysis, PMRDT (P<0.0001) and age >15 years (P=0.002) were associated with higher TRM, active disease (P=0.0021), ABMT (P=0.0074) and male sex (P=0.011) with higher relapse, and age >15 years (P=0.0007) and PMRDT (P=0.047) with lower DFS. Amongst second remission patients, TRM was higher after PMRDT (P=0.0003), relapse was higher after ABMT (P=0.034), and 5-year DFS was comparable (32% ABMT and 25% PMRDT). ABMT, if feasible, may be preferable to PMRDT in advanced acute leukemia patients since lower relapse after PMRDT is offset by higher TRM. If an autograft is not feasible because of nonavailability of autologous cells or very advanced disease, PMRDT is a potential alternative. PMID- 12748666 TI - Role of pretransplant interferon-alpha(IFN) treatment in the outcome of stem cell transplantation (SCT) from related donors in chronic myelogenous leukemia (CML): results from three Turkish transplant centers. AB - Since transplantation cannot be performed immediately after the diagnosis of chronic myelogenous leukemia (CML), interferon treatment is usually required. This study aims to analyze the effects of interferon-alpha (IFN) treatment on allogeneic stem cell transplantation (SCT) outcome. A total of 106 patients aged 16-47 years and transplanted from HLA-identical sibling donors for CML in chronic phase (CP) were evaluated. In all, 48 had received IFN-alpha for a median duration of 5 months (1-18 months) until a median of 1 month prior to transplantation. Of the patients, 50 have received bone marrow transplant (BMT) whereas 56 have received peripheral blood stem cells (PBSCT) between 1991 and 1999 in three major transplant centers in Turkey. Patient characteristics in both groups were similar. More hematological responders were present in the IFN(+) patients (P=0.0001). No difference was found in engraftment kinetics. The incidences of acute or chronic graft-versus-host disease (GVHD), relapse and graft failure were similar in all patients regardless of stem cell source. Overall survival (OS) and disease-free survival (DFS) at 2 years were similar for both IFN(+) or (-) patients following SCT. With multivariate analysis, pretransplant IFN-alpha use, stem cell source, transplant year and CD34+ cell content were not found to be risk factors for OS. In conclusion, prior IFN exposure did not impair BMT or PBSCT outcome. PMID- 12748667 TI - Prognostic value of cyclic GMP in patients undergoing allogeneic bone marrow transplantation after conditioning with total body irradiation. AB - This study was performed to investigate whether measurement of cyclic GMP (cGMP), a marker for nitric oxide production, before and after allogeneic bone marrow transplantation (BMT) with total body irradiation (TBI) conditioning was of prognostic value. cGMP levels were monitored in 23 consecutive patients who received TBI as conditioning for BMT, and were compared with the outcome. cGMP became positive during the aplastic phase after BMT in 12 patients. In nine of these 12 patients, cGMP level decreased during the recovery phase. Eight of the nine patients survived, one dying after relapse. In three other patients, the cGMP level continued to increase even during the recovery phase and they died of severe complications. cGMP became positive on day 0 of BMT and during the leukocyte recovery phase after BMT in two and seven of the 23 patients, respectively. Subsequently, all patients died of severe complications. The two patients who were negative for cGMP both before and after BMT survived without complications. These results suggest that monitoring cGMP from early after BMT may be useful for predicting outcome and that it may be a useful prognostic marker. PMID- 12748668 TI - A potential graft-versus-leukemia effect after allogeneic hematopoietic stem cell transplantation for patients with Philadelphia chromosome-positive acute lymphoblastic leukemia: results from the French Bone Marrow Transplantation Society. AB - SUMMARY: Philadelphia chromosome (Ph) acute lymphoblastic leukemia-positive (ALL) is a subgroup of ALL with a poor prognosis. We sought to evaluate the results of allogeneic hematopoietic stem cell transplantation (HSCT) in this situation. From 1992 to 2000, 121 patients with Ph- positive ALL enrolled in one of the three main French prospective ALL chemotherapy trials and receiving allogeneic HSCT were reported to the registry of the French Society of Bone Marrow Transplantation (SFGM-TC). The median age was 35 years (range, 1-53). In all, 76 patients received HSCT in first complete remission and 45 in a more advanced disease stage. Minimal residual disease was evaluated just before the graft: 35 patients of the 52 patients in first remission had persistent BCR-ABL transcript detectable while 17 had no detectable minimal residual disease. Bone marrow and/or peripheral blood samples from 94 patients were submitted for reverse transcriptase polymerase chain reaction analysis at variable points after transplantation. Estimated 2-year survival and relapse rate for patients in CR1 were 50 and 37%, respectively, significantly better than for patients with more advanced disease (P=0.0001 and 0.01, respectively). There was no difference in survival or in relapse rates in terms of the donor used. Relapse was the most common cause of treatment failure. Hematological status at the time of transplant and the occurrence of acute graft-versus-host disease (GvHD) were the only two prognostic factors identified for relapse (P=0.02 and 0.02, respectively). Detection of BCR-ABL transcripts after transplantation had a predictive value on relapse occurrence. Finally, donor lymphocyte infusions were successfully used to treat some relapses. The graft-versus-leukemia effect of HSCT in Ph-positive ALL appears to be supported by (1) the lack of prognostic significance of pretransplant BCR-ABL transcript detection, (2) the efficacy of donor lymphocyte infusions in cases of relapse, and (3) the role of GvHD as protecting against relapse. PMID- 12748670 TI - Successful treatment of severe hemorrhagic cystitis after hemopoietic cell transplantation by selective embolization of the vesical arteries. AB - Hemorrhagic cystitis (HC) is a common and sometimes life-threatening complication of hemopoietic cell transplantation (HCT) occurring in 7-52% of transplant recipients. In this setting it is usually either related to the use of cyclophosphamide or to a viral infection (BK, JC viruses and adenovirus type 11). Treatment is based on hyperhydration, platelet and blood-cell transfusions, bladder irrigation and pain management. Where these measures have failed to control HC, numerous therapeutic approaches including surgery have been tried with poor success. We report two HCT patients with severe HC successfully treated with selective embolization of the vesical arteries. PMID- 12748669 TI - Mesial temporal sclerosis--a late complication in four allogeneic pediatric recipients with persistent seizures after an acute episode of cyclosporine-A neurotoxicity. AB - Mesial temporal sclerosis (MTS) is a common finding in patients with intractable temporal lobe epilepsy (TLE). In this report, we retrospectively reviewed the neuroimaging results of four children who underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT), and who developed recurrent, partial, intractable seizures following a first event caused by cyclosporine-A (CSA) neurotoxicity. Neuroradiologic findings of MTS were demonstrated in all these patients. We suggest that MTS may be a consequence of CSA neurotoxicity, which induces repeated seizures, associated with other predisposing conditions, as well as being a consequence of the underlying disease and its treatment, and of severe graft-versus-host disease (GvHD). PMID- 12748671 TI - Central venous catheter pinch-off and fracture: recognition, prevention and management. AB - The so-called pinch-off syndrome is observed in up to 1% of all central venous catheters (CVCs), and is a valuable warning prior to fragmentation, which occurs in approximately 40% of the respective cases. As long-term indwelling CVCs are used with increasing frequency, this paper describes the necessity of pinch-off monitoring following the experiences of a case study and a review of the current literature on this specific topic in order to point out preventive practice guidelines. Besides easy preventive practices such as a high level of suspicion and adequate X-ray controls, findings give strong evidence that the most important specific factor might be the adequate approach. In our hands, the supraclavicular technique has provided the best results with regards to percutaneous introduction of large bore CVCs. PMID- 12748672 TI - B-cell lymphoma developing in the donor 9 years after donor-origin acute myeloid leukemia post bone marrow transplantation. AB - Donor-cell leukemia post bone marrow transplantation is a rare event. Most of the cases reported to date have developed in cells from an HLA-matched sibling, who had no evidence of malignant disease before or following the occurrence of donor origin leukemia. We describe a 17-year-old female who developed B-cell lymphoma 9 years following the occurrence of donor-origin acute myeloid leukemia in her brother for whom she had donated marrow. Cytogenetic analysis of the tumor revealed multiple chromosomal aberrations. The donor was heterozygous for the Ashkenazi mutation of Bloom's syndrome, suggesting that donor-type leukemia could have resulted from genomic instability in the donor cells. PMID- 12748673 TI - Peracute onset of severe tumor lysis syndrome immediately after 4 Gy fractionated TBI as part of reduced intensity preparative regimen in a patient with T-ALL with high tumor burden. AB - We report a 30-year-old patient with therapy-refractory T-ALL undergoing unrelated allogeneic PBSCT. He developed severe tumor lysis syndrome (TLS) with extreme biochemical changes, cardiac and neurological symptoms and dialysis dependent acute renal failure after TBI (4 Gy) on the first day of reduced intensity conditioning (RIC) for unrelated allogeneic PBSCT. The patient's clinical condition was stabilized after beginning daily hemodialysis and treatment for disturbed electrolytes, metabolic acidosis and plasma coagulation, as well as reduction of uric acid by rasburicase. The conditioning therapy and the allogenic PBSCT were scheduled according to the preparative regimen. According to our knowledge, severe TLS induced by 4 Gy TBI has not been reported so far. Regimen-related toxicity using RIC regimen was mild, allowing 30-50% of the patients to have an entirely outpatient transplantation. However, we would like to point out that severe TLS could also complicate PBSCT using RIC regimens in patients with relatively radiation-sensitive malignancies and high tumor burden. PMID- 12748675 TI - Regression of an unresectable pancreatic tumor following nonmyeloablative allogeneic peripheral-blood stem-cell transplantation. AB - A 59-year-old female with an unresectable, large pancreatic tumor (10.0 x 8.0 cm(2) on CT scan) underwent nonmyeloablative allogeneic peripheral-blood stem cell transplantation from her HLA-identical sibling. Pronounced tumor regression and relief from pain without acute graft-versus-host disease (GVHD) were observed following transplantation. The patient is surviving (more than 300 days) after transplantation, with extensive chronic GVHD, and has tumor regression with an 80% reduction in tumor size. The observed clinical course may suggest a graft versus-tumor effect on the pancreatic tumor following allogeneic stem-cell transplantation. PMID- 12748674 TI - Interferon-gamma for delayed pulmonary toxicity syndrome resistant to steroids. AB - Delayed pulmonary toxicity syndrome, characterized by interstitial pneumonia and pulmonary fibrosis, is common following high-dose bischloroethylnitrosourea (BCNU) (carmustine, [1,3-bis (2-chloroethyl)-1-nitrosourea]) containing chemotherapeutic regimens. Depending upon the treatment protocol, it may develop in over 70% of patients. Early and aggressive corticosteroid treatment leads to improvement in the majority of patients. However, up to 8% of affected patients may fail to respond to corticosteroids and develop progressive respiratory failure leading to death. No alternatives to corticosteroids have thus far been shown useful. We report the symptomatic and physiological improvement of a patient with severe steroid-resistant delayed pulmonary toxicity syndrome, following treatment with interferon-gamma. PMID- 12748676 TI - Concurrent presence of both patient and donor t(14;18) in a follicular lymphoma patient after undergoing allogeneic BMT: implications for minimal residual disease detection post-transplant. AB - We report the case of a t(14:18)(+) follicular lymphoma (FL) patient in long-term clinical remission after undergoing an allogeneic bone marrow transplantation (allo-BMT) from a human leukocyte antigen (HLA)-identical sibling donor who was the normal healthy carrier of a t(14:18)(+) B cell clone. Using real-time quantitative PCR (RQ-PCR) and gel electrophoresis, we document the temporal disappearance of the patient's t(14:18)(+) clone early post-transplant with the concomitant emergence and long-term persistence of the donor's t(14:18)(+) clone in the patient's peripheral blood. This report indicates that the use of PCR based techniques to measure minimal residual disease in FL patients post-alloBMT should incorporate pretransplant screening of the donor for t(14;18). Furthermore, it suggests that healthy individuals with t(14:18) need not be excluded as donors for FL patients treated with allo-BMT. PMID- 12748678 TI - NTP Toxicology and Carcinogenesis Studies of Benzyl Acetate (CAS No. 140-11-4) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - Benzyl acetate, a water-white liquid with a pear-like odor, is a natural constituent of several essential oils and flower absolutes extracted from jasmine, hyacinth, gardenia, tuberose, ylang-ylang, cananga, and neroli. Commercial benzyl acetate, a liquid prepared synthetically from benzyl chloride, acetic acid, and triethylamine is used primarily as a component of perfumes for soaps and as a flavoring ingredient. This compound is practically insoluble in water but is miscible in alcohol and ether and soluble in benzene and chloroform. Toxicology and carcinogenesis studies of benzyl acetate (>99% pure) were conducted by administering benzyl acetate in corn oil gavage to groups of 50 male and 50 female F344/N rats at doses of 0, 250, or 500 mg/kg body weight and to groups of 50 male and 50 female B6C3F1 mice at doses of 0, 500, or 1,000 mg/kg once daily five days per week for 103 weeks. Dose selection for the 2-year study was based on mean body weight gain depression and decreased survival observed at higher doses in 13 week studies. The absence of any observable adverse effect of benzyl acetate on the survival or mean body weight gains of the rats or mice in the 2-year studies suggests that both the rats and the mice of each sex could have tolerated higher doses. An infection in the genital tract was probably responsible for the deaths of 26/35 control, 14/32 low-dose, and 8/20 high-dose female mice before the end of the study. Acinar-cell adenomas in the pancreas of male rats occurred with a positive trend (P<0.01), and the incidence in the high dose group (37/49, 76%) was significantly (P<0.01) higher than in the vehicle controls (22/50, 40%). The incidence of these tumors in the low-dose group (27/50, 54%) was comparable to that in the gavage controls. Acinar-cell hyperplasia of the pancreas was observed in 37/50 control, 34/50 low-dose, and 36/49 high-dose male rats. No acinar-cell hyperplasia or adenoma of the pancreas was observed in female rats. The incidence of retinopathy and cataracts in the high-dose male rats was increased compared with the controls (retinopathy: 1/50; 0/50; 20/50; cataracts: 0/50; 0/50; 13/50). Low-dose female rats had an increased incidence of retinopathy (18/50). Retinopathy and cataracts in rats have been associated with proximity to fluorescent light in this and previous studies. Preputial gland neoplasms occurred with a positive trend (P<0.05) in male rats (cystadenocarcinoma: 0/50; 0/50; 3/50; all adenocarcinoma: 0/50; 1/50; 4/50; adenocarcinoma or carcinoma combined: 1/50; 1/50; 6/50). However, the incidence of all preputial gland tumors was not significantly elevated (2/50; 1/50; 6/50). For female rats the incidence of clitoral gland neoplasms was marginally increased (2/50; 0/50; 5/50). Hepatocellular adenomas occurred in mice of each sex with statistically positive trends (males: 0/50; 5/49; 13/50; females: 0/50; 0/50; 6/50), and the incidences in the high-dose groups were greater than those in the controls (males: P<0.001; females: P<0.05). Hepatocellular carcinomas were marginally elevated in dosed male and high-dose female mice (males: 10/50; 14/49; 12/50; females: 1/50; 0/50; 4/50). Squamous cell papillomas or carcinomas of the forestomach (uncommon neoplasms) occurred with a positive trend (P<0.05) in male mice (4/49; 4/48; 11/49). The incidence of these tumors was also marginally (P=0.054) increased in the high-dose female mice (0/50; 0/50; 4/48). The incidences of these tumors in both the high-dose male and the high-dose female mice were considerably higher than the historical corn oil gavage control rates at this laboratory (males, 2/296, 0.7%; females, 2/297, 0.7%) and throughout the program (males, 14/1,070, 1.3%; females, 3/1,073, 0.3%). Forestomach hyperplasia occurred at increased incidences in dosed mice of either sex (males: 1/49, 7/48, 22/49; females: 1/50, 6/50, 17/48). The neoplasms and hyperplasia of the forestomach were probably related to administration of benzyl acetate. In a separate metabolism study, benzyl acetate was absorbed from the gastrointestinal traolism study, benzyl acetate was absorbed from the gastrointestinal tract of rats and mice, with approximately 90% of the administered dose recovered as various metabolites in the urine within 24 hr. The primary metabolite was hippuric acid, with minor amounts of a mercapturic acid, and one or more unidentified metabolites. This capacity for absorption, metabolism, and disposition was unaffected by the amount or number of doses administered. Benzyl acetate was not mutagenic in strains TA100, TA98, TA135, or TA137 of Salmonella typhimurium in the presence or absence of Aroclor 1254-induced Sprague-Dawley rat or Syrian hamster S9 when tested according to the preincubation protocol. Benzyl acetate did not induce sister-chromatid exchanges or chromosomal aberrations in Chinese hamster ovary cells in the presence or absence of Aroclor 1254-induced Sprague-Dawley rat liver S9. Benzyl acetate was mutagenic in the mouse lymphoma L5178Y/TK± assay in the presence, but not in the absence, of Aroclor 1254 induced Fisher 344 rat liver S9. An audit was conducted on the experimental data and the draft technical report for these 2-year studies on benzyl acetate. Based on the results of this audit additional pathology examinations were conducted on all target organs in male rats and male and female mice. The Technical Report reflects these final pathology evaluations. The overall conclusions regarding the toxicology and carcinogenicity of benzyl acetate did not change as a result of this evaluation. Under the conditions of these gavage studies, benzyl acetate increased the incidence of acinar-cell adenomas of the exocrine pancreas in male F344/N rats; the gavage vehicle may have been a contributing factor. There was no evidence of carcinogenicity for female F344/N rats. For male and female B6C3F1 mice there was some evidence of carcinogenicity in that benzyl acetate caused increased incidences of hepatocellular adenomas and squamous cell neoplasms of the forestomach. Synonyms: alpha-acetoxytoluene; benzyl ethanoate; acetic acid, benzyl ester PMID- 12748677 TI - Epstein-Barr virus-associated B-cell lymphoproliferative disorder in CLL patients after treatment with fludarabine and cyclophosphamide followed by high-dose chemotherapy with autologous stem cell transplantation. PMID- 12748679 TI - NTP Lifetime Carcinogenesis Studies of Amosite Asbestos (CAS No. 12172-73-5) in Syrian Golden Hamsters (Feed Studies). AB - Carcinogenesis studies of amosite asbestos were conducted by administering diets containing 1% of the asbestos in pellets from the conception of the mothers through the lifetime of male and female Syrian golden hamsters. Control groups consisted of 127 male and 126 female hamsters and the amosite asbestos groups consisted of 252 male and 254 female hamsters. No adverse effect on body weight gain or survival was observed from treatment with amosite asbestos. Neither of the amosite asbestos groups showed increased neoplasia in any organ or tissue compared to the control groups. Under the conditions of these studies, the ingestion of amosite asbestos at a level of 1% in the diet for their lifetime was not toxic and did not cause a carcinogenic response in male and female Syrian golden hamsters. Levels of Evidence of Carcinogenicity: Male Hamsters: Negative Female Hamsters: Negative PMID- 12748680 TI - NTP Toxicology and Carcinogenesis Studies of Tris(2-ethylhexyl)phosphate (CAS No. 78-42-2) In F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - Tris(2-ethylhexyl)phosphate is one of a family of triakyl phosphates that have been widely used as fire retardants and plasticizers. Another triakyl phosphate, tris(2,3-dibromopropyl)phosphate (Tris-BP), once used as a flame retardant in children's sleepwear, has been shown to be carcinogenic, but tris(2 ethylhexyl)phosphate has not been previously studied. Tris(2 ethylhexyl)phosphate, a clear, viscous liquid, is used as a component of vinyl stabilizers, grease additives, and flame-proofing compositions; however, it is used primarily as a plasticizer for vinyl plastic and synthetic rubber compounds. In 1974, approximately 3 million pounds of tris(2-ethylhexyl)phosphate was produced in the United States; imports during that year were negligible. Substantial human exposure probably occurs during production of tris(2 ethylhexyl)phosphate and during the manufacture and use of products containing it, but data on the magnitude of exposure are not available. Two-year toxicology and carcinogenesis studies of tris(2-ethylhexyl)phosphate were conducted by administering the test chemical in corn oil by gavage, 5 days per week for 103 weeks, to groups of 50 male and 50 female F344/N rats and B6C3F1 mice. Male rats received doses of 2,000 or 4,000 mg/kg body weight, female rats received 1,000 or 2,000 mg/kg, and male and female mice received 500 or 1,000 mg/kg. Fifty vehicle control animals of each sex and species received 10 ml/kg body weight (rats) or 3.3 ml/kg (mice) corn oil by gavage on the same schedule. Inflammation of the gastric mucosa in mice and mild weight depression in rats and mice were the only dose-related effects observed in the preliminary studies. In the 2-year studies, survival rates and mean body weight gains of dosed female rats and dosed mice were comparable to those of their perspective controls. Survival rates of dosed male rats were comparable to that of the vehicle controls, but body weight gains were depressed. One nonneoplastic lesion, follicular cell hyperplasia of the thyroid, was observed at increased incidences in dosed male and female mice. Two compound-related increased incidences of neoplasms could not be discounted. In male rats, the incidence of pheochromocytoma of adrenal glands increased with dose (2/50, 4%; 9/50, 18%; 12/50, 24%). There were also two additional malignant pheochromocytomas in the high dose group. However, the incidence of adrenal pheochromocytoma in vehicle controls of this study (2/50, 4%) was low compared with the 25% incidence observed in two previous studies in this laboratory or the overall historical incidence of 18% observed throughout the Program, and thus the evidence of carcinogenicity was considered to be equivocal. In female mice, the incidence of hepatocellular carcinoma (0/48; 4/50; 7/50) in high dose animals (1,000 mg/kg) was significantly increased relative to that of the vehicle controls. Decreased incidences were observed for acinar cell adenomas of the pancreas in dosed male rats (14/50, 28%; 5/48, 10%; 2/49, 4%) and for fibroadenomas of the mammary glands in low dose female rats (11/50, 22%; 2/50, 4%; 7/50, 14%). Hemangiosarcomas of the circulatory system in male mice (7/50, 14%; 0/50; 1/49, 2%) and lymphomas of the hematopoietic system in female mice (14/49, 29%; 10/50, 20%; 6/50, 12%) were decreased compared with vehicle controls. A decrease in the incidence of lymphomas and an increased incidence of carcinomas of the liver in female mice (both seen in this study) were observed in studies of di(2-ethylhexyl)adipate. Increased incidences of liver carcinomas and decreased incidences of mammary fibroadenomas were observed also in female rats in the di(2-ethylhexyl)phthalate studies. A possible link among these three chemicals may be metabolic conversion to 2-ethylhexanol. Tris(2 ethylhexyl)phosphate was not mutagenic in Salmonella typhimurium strains TA98, TA100, TA1535, or TA1537 in the presence or absence of 9000 x g (S9) fractions from Aroclor 1254-induced Sprague-Dawley rat or Syrian hamster liver. An audit of the experimental data from these carcinogeneoclor 1254-induced Sprague-Dawley rat or Syrian hamster liver. An audit of the experimental data from these carcinogenesis studies was conducted by the National Toxicology Program. No data discrepancies were found that significantly influenced the final interpretations of these experiments. Under the conditions of these studies, a comparison of concurrent and historical controls indicated that there was equivocal evidence of carcinogenicity in male F344/N rats receiving 2,000 and 4,000 mg/kg tris(2 ethylhexyl)phosphate, as evidenced by increased incidences of pheochromocytomas of the adrenal glands. There was no evidence of carcinogenicity in female F344/N rats or in male B6C3F1 mice receiving tris(2-ethylhexyl)phosphate. There was some evidence of carcinogenicity in female B6C3F1 mice that received 1,000 mg/kg tris(2-ethylhexyl)phosphate, as shown by an increased incidence of hepatocellular carcinoma. Tris(2-ethylhexyl)phosphate was associated with increased incidences of follicular cell hyperplasia of the thyroid gland in male and female B6C3F1 mice. Synonyms and Trade Names: TOF; trioctyl phosphate; phosphoric acid tri(2 ethylhexyl) ester; Flexolreg. TOF; Kronitexreg. PMID- 12748681 TI - NTP Toxicology and Carcinogenesis Studies of Trichloroethylene (CAS No. 79-01-6) in Four Strains of Rats (ACI, August, Marshall, Osborne-Mendel) (Gavage Studies). AB - Trichloroethylene is an industrial solvent used primarily for vapor degreasing and cold cleaning. It was selected for study because of its industrial use and for potential for human exposure. (An estimated 3.5 million workers are exposed to trichloroethylene.) In an earlier study trichloroethylene (stabilized with epichlorohydrin and 1,2-epoxybutane) administered by gavage caused hepatocellular carcinomas in male and female B6C3F1 mice. Trichloroethylene administration did not increase the incidence of tumors in male or female Osborne-Mendel rats. However, the survival of dosed rats was reduced, thereby compromising the sensitivity of the study to detect a carcinogenic effect. The studies described in this report were conducted to compare the sensitivities of four strains of rats (ACI, August, Marshall, and Osborne-Mendel) to diisopropylamine-stabilized trichloroethylene. The results of the present studies demonstrate that long-term administration of trichloroethylene produces nephrotoxicity in four strains of rats and that the susceptibilities of these strains to the nephrotoxic effects of the chemical are similar. Because of chemically induced toxicity, reduced survival, and incomplete documentation of the experimental data, the studies are considered inadequate for either comparing or assessing trichloroethylene-induced carcinogenesis in these strains of rats. Toxicology and carcinogenesis studies of trichloroethylene (more than 99% pure, stabilized with 8 ppm diisopropylamine) were conducted by administering the chemical in corn oil gavage at doses of 0, 500, or 1,000 mg/kg per day, 5 day per week, for 103 weeks to groups of 50 male and 50 female ACI, August, Marshall, and Osborne-Mendel rats. The doses were selected on the basis of results from 13-week gavage studies in which groups of 10 male and 10 female ACI, August, and Marshall rats received daily doses or trichloroethylene (male: 125-2,000 mg/kg; female: 63-1,000 mg/kg). Doses for Osborne-Mendel rats were selected to conform with doses used in an earlier carcinogenicity study in that strain (TR-2). In the 13-week studies, male ACI and August rats receiving 2,000 mg/kg trichloroethylene and male and female Marshall rats receiving 1,835 mg/kg had final mean body weights 12%-17% lower than those of the vehicle controls. All other dose groups had body weights comparable to those of the vehicle controls. Three male August rats dosed with 2,000 mg/kg died. Histopathologic evaluation of tissues revealed no lesions attributable to trichloroethylene administration in the 13-week studies. This absence of histopathologic findings did not accurately predict the nephrotoxic effects of long-term administration of trichloroethylene to rats. Body Weight and Survival in the Two-Year Studies: In the 2-year studies, all dosed groups exhibited some reduction in mean body weights relative to the vehicle controls. Survival relative to vehicle controls was significantly reduced in 7/16 dosed groups (see page 6 of the Technical Report). Also, the survival of high dose male Marshall rats was reduced by a large number of accidental deaths. Nephrotoxicity, reduced survival, and central nervous system toxicity (characterized by sedation, loss of consciousness, tremors, and convulsions) showed that the doses of trichloroethylene selected for the 2-year studies were too high. Renal Effects in the Two-Year Studies: Trichloroethylene caused tubular cell cytomegaly in 82% 100% of all dosed animals. In addition, trichloroethylene produced toxic nephropathy (which was distinguishable from age-related nephropathy) in 17%-80% of the dosed animals. Cytomegaly, karyomegaly, or toxic nephropathy was not found in untreated or vehicle control animals. Trichloroethylene administration was also associated with increased incidences of renal tubular cell adenomas and adenocarcinomas. The incidences of renal lesions are shown in the following table (see page 7 of Technical Report). Other Pathologic Effects in the Two-Year Studies: An increased incidence of interstitial cell tumors of the testis was observinterstitial cell tumors of the testis was observed in high dose male Marshall rats (untreated control, 16/46; vehicle control, 17/46; low dose, 21/48; high dose, 32/48; P=0.002). The incidences of pheochromocytomas of the adrenal gland were significantly reduced in male ACI, female August, female Marshall, and male and female Osborne-Mendel rats. Genetic Toxicology: Trichloroethylene did not cause mutations in Salmonella typhimurium strains TA98, TA100, TA1535, or TA1537 with or without metabolic activation. In Chinese hamster ovary cells, trichloroethylene did not induce chromosomal aberrations; the results for sister chromatid exchanges were considered positive. Trichloroethylene was mutagenic to mouse L5178Y lymphoma cells in the presence of rat liver S9. Data Audit: Audits of the experimental data for these 2-year studies of trichloroethylene were conducted by the National Toxicology Program (see Appendix Q of the Technical Report). The results of the audits revealed evidence that the doses of trichloroethylene were too high. In addition, there was insufficient documentation of animal breeding, clinical observations, environmental conditions, and analytical chemistry data. Also, individual animal identification was not always verifiable. Conclusions: Under the conditions of these 2-year gavage studies of trichloroethylene in male and female ACI, August, Marshall, and Osborne-Mendel rats, trichloroethylene administration caused renal tubular cell cytomegaly and toxic nephropathy in both sexes of the four strains. However, these are considered to be inadequate studies of carcinogenic activity because of chemically induced toxicity, reduced survival, and deficiencies in the conduct of the studies. Despite these limitations, tubular cell neoplasms of the kidney were observed in rats exposed to trichloroethylene and interstitial cell neoplasms of the testis were observed in Marshall rats exposed to trichloroethylene. Synonyms: acetylene trichloride; 1-chloro-2,2-dichloroethylene; 1,1-dichloro-2 chloroethylene; ethinyl trichloride; ethylene trichloride; 1,1,2 trichloroethylene; trichloroethene Trade names of formulations: Algylen; Anamenth; Benzinol; Blacosolv; Blancosolv; Cecolene; Chlorilen; Chlorylea; Chorylen; Circosolv; Crawhaspol; Densinfluat; Dow-Tri; Dukeron; Fleck-Flip; Flock Flip; Fluate; Gemalgene; Germalgene; Lanadin; Lethurin; Narcogen; Narkogen; Narkosoid; Nialk; Perma-A-Chlor; Perm-A-Clor; Petzinol; Philex; Threthylen; Threthylene; Trethylene; Tri; Triad; Trial; Triasol; Trichloran; Trichloren; Triclene; Tri-Clene; Trielene; Trielin; Triklone; Trilen; Trilene; Triline; Trimar; Triol; TRI-plus; TRI-plus M; Vestrol; Vitran; Westrosol Target Organs & Incidences from 2-year Studies PMID- 12748682 TI - NTP Toxicology and Carcinogenesis Studies of Propylene (CAS No. 115-07-1) in F344/N Rats and B6C3F1 Mice (Inhalation Studies). AB - Propylene is used as a starting material in the production of polypropylene plastics and various other chemicals, including acrylonitrile, isopropyl alcohol, propylene oxide, butyraldehyde, cumene, dodecane, nonene, and allyl chloride. The major derivatives are polypropylene (25%), acrylonitrile (15%), isopropyl alcohol (10%), and propylene oxide (10%). It is also a valuable feed-stock chemical for the production of gasoline. Other miscellaneous applications include use as a starting material for polymerization reactions to form vinyl chloride copolymers and low-molecular-weight homopolymers that are used as additives in lubricating oils and in the manufacture of hydroquinone. The chemical is also used as an aerosol propellant or component. The major end uses of propylene are in the production of fabricated plastics (50%) and fibers (15%). Toxicology and carcinogenesis studies of propylene (greater than 99% pure) were conducted by exposing groups of 50 F344/N rats and 49 or 50 B6C3F1 mice of each sex to propylene in air by inhalation at concentrations of 5,000 or 10,000 ppm, 6 hours per day, 5 days per week, for 103 weeks. Other groups of 50 rats and 50 mice of each sex in chambers received air only on the same schedule and served as chamber controls. The highest concentration of propylene that was considered safe for these studies was 10,000 ppm because of the risk of explosion that can occur at higher concentrations. The survival of exposed and control rats and mice was comparable. Throughout most of the studies, mean body weights of exposed male and female rats were slightly lower (0%-5%) than those of the controls, but the decrements were not concentration related. After week 59 of the study, mean body weights of 10,000-ppm male mice were usually slightly lower (5%) than those of the controls, whereas those in other exposed groups of male and female mice were generally comparable with those of the controls. No compound-related adverse clinical signs were observed in either species. An increased incidence of squamous metaplasia of the nasal cavity was observed in female rats exposed at the 5,000-ppm and 10,000-ppm concentrations (control, 0/49; low, 15/50; high, 6/50) and in male rats exposed at 5,000 ppm (2/50; 19/50; 7/50). Epithelial hyperplasia of the nasal cavity was increased in female rats exposed at the 10,000-ppm concentration (0/49; 4/50; 9/50); the incidences in male rats were 2/50, 2/50, and 5/50. Inflammation of the nasal cavity, characterized by an influx of lymphocytes, macrophages, and granulocytes into the submucosa and by granulocytes into the lumen, occurred at increased incidences in low concentration and high concentration male rats and in high concentration female rats. Chronic focal inflammation of the kidneys occurred at an increased incidence in low concentration and high concentration mice of each sex. Hemangiosarcomas were found in one low dose male mouse (liver), two high dose male mice (spleen), and three high dose female mice (subcutis, spleen, and uterus). Hemangiomas were found in one low dose and in one high dose female mouse (liver). Vascular tumors were not found in control mice of either sex. The low incidences of vascular tumors and their occurrence in a variety of organs suggest that they are not related to administration of propylene. The occurrence of uterine endometrial stromal polyps in female mice showed a positive trend (P<0.05; 0/47; 0/47; 3/48); the incidence in the 10,000-ppm group was not significantly greater than that in the concurrent control group, but the incidence was higher than the mean historical control rate (22/2,411, 0.9%) and was within the range (0%-6%) observed in studies throughout the Carcinogenesis Program. The occurrence of endometrial stromal polyps in three high concentration female mice was not considered to be clearly related to exposure to propylene. The incidence of male mice with alveolar/bronchiolar adenomas or carcinomas (combined) occurred with a negative trend (P<0.05; 16/50; 4/49; 7/50), and the reduced incidences in both exposed groups wegative trend (P<0.05; 16/50; 4/49; 7/50), and the reduced incidences in both exposed groups were less (P<0.05) than that in the control group. The control incidence of these tumors in an inhalation study conducted concurrently at the same laboratory was similar (15/50), suggesting a possible exposure-related decrease. The biologic significance of this decrease in male mice is difficult to assess; the incidences seen in these control and exposed animals are within the range of incidences (2% 34%; mean, 16.7%) observed in control male mice in other studies throughout the Carcinogenesis Program. An audit of the experimental data was conducted for these carcinogenesis studies on propylene. No data discrepancies were found that influenced the final interpretations. Under the conditions of these studies, there was no evidence of carcinogenicity in male and female F344/N rats or in male and female B6C3F1 mice exposed to propylene by inhalation at concentrations of 5,000 or 10,000 ppm for 103 weeks. In the nasal cavity, propylene induced squamous metaplasia of the respiratory epithelium in male and female rats and epithelial hyperplasia in female rats. Synonyms: propene; methylethylene; methylethene PMID- 12748683 TI - NTP Toxicology and Carcinogenesis Studies of HC Blue No. 1 (CAS No. 2784-94-3) in F344/N Rats and B6C3F1 Mice (Feed Studies). AB - Toxicology and carcinogenesis studies of HC Blue No. 1 (97% pure), a semipermanent hair dye, were conducted by administering the test chemical in feed for 103 weeks to groups of 50 F344/N rats and 50 B6C3F1 mice of each sex. The dietary concentrations used were 0, 1,500, or 3,000 ppm for rats and male mice and 0, 3,000, or 6,000 ppm for female mice. These concentrations were selected on the basis of results from single-administration gavage studies and 14-day and 13 week feed studies. The survival of male and female rats and male mice was not affected by administration of HC Blue No. 1. Survival of high dose female mice was reduced (P<0.05); the early deaths in this group are believed to have been caused by hepatocellular carcinomas. Body weights of high dose rats and dosed mice were lower than those of the respective control groups; female rats and mice were more affected than were males. Administration of HC Blue No. 1 produced significant positive trends in the incidences of male rats with hepatocellular neoplastic nodules/carcinomas (neoplastic nodules: control, 0/49; low dose, 0/50; high dose, 3/50; neoplastic nodules/carcinomas: 1/49; 0/50; 6/50). In male and female mice, both doses of HC Blue No. 1 increased the incidences of hepatocellular carcinoma (male: 11/50; 20/50; 30/50; female: 1/50; 24/48; 47/49) and the low doses increased the incidences of hepatocellular adenomas (male: 4/50; 17/50; 10/50; female: 2/50; 11/48; 4/49). HC Blue No. 1 produced dose related increases in the incidences of proliferative lesions of the lungs (adenomatous hyperplasia and alveolar/bronchiolar adenomas or carcinomas) in female rats (hyperplasia: 2/50; 5/49; 8/50; adenoma/carcinoma: 1/50; 3/49; 7/50). In male mice, HC Blue No. 1 at the 6,000-ppm dose increased the incidences of thyroid gland follicular cell hyperplasia and adenomas (hyperplasia: 3/47; 7/49; 14/50; adenoma: 0/47; 0/49; 5/50). HC Blue No. 1 was mutagenic in strains TA97, TA98, and TA100 of Salmonella typhimurium in the presence or absence of Aroclor induced male Sprague-Dawley rat or Syrian hamster liver S9; HC Blue No. 1 was negative in strain TA1535. HC Blue No. 1 was mutagenic in the absence of activation in the L5178Y/TK+/- mouse lymphoma assay and induced unscheduled DNA synthesis in rat hepatocytes in vitro. An audit of the experimental data was conducted for these carcinogenesis studies on HC Blue No. 1. No data discrepancies were found that influenced the final interpretations. Under the conditions of these feed studies, there was equivocal evidence of carcinogenicity in male F344/N rats, since HC Blue No. 1 caused a marginal increase in the incidence of hepatocellular neoplastic nodules/carcinomas. For female F344/N rats, there was some evidence of carcinogenicity in that HC Blue No. 1 induced increased incidences of alveolar/bronchiolar neoplasms. There was clear evidence of carcinogenicity of HC Blue No. 1 for male and female B6C3F1 mice as shown by increased incidences of hepatocellular carcinomas. The incidences of follicular cell adenomas of the thyroid gland were also increased in male mice receiving HC Blue No. 1. Synonym: 2,2'((4-(methylamino)-3-nitrophenyl)imino)bis (ethanol) PMID- 12748684 TI - NTP Toxicology and Carcinogenesis Studies of Telone II(R) (Technical-Grade 1,3 Dichloropropene [CAS No. 542-75-6] Containing 1.0% Epichlorohydrin as a Stabilizer) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - Toxicology and carcinogenesis studies of Telone II(R) (a soil fumigant containing approximately 89% cis- and trans-1,3-dichloropropene, 2.5% 1,2-dichloropropane, 1.5% of a trichloropropene isomer, and 1.0% epichlorohydrin) were conducted by administering the commercial-grade formulation in corn oil by gavage to groups of 52 male and 52 female F344/N rats at doses of 0, 25, or 50 mg/kg and to groups of 50 male and 50 female B6C3F1 mice at doses of 0, 50, or 100 mg/kg. Doses were administered three times per week for 104 weeks. Ancillary studies were conducted in which dose groups containing five male and five female rats were killed after receiving Telone II(R) for 9, 16, 21, 24, or 27 months. Mean body weights of high dose male rats were about 5% lower than those for the vehicle control and low dose male rats; no differences in body weights were observed among groups of female rats. Survival was comparable for the different groups of male and female rats. For male and female mice, the dosed groups initially weighed 6%-22% less than did the vehicle controls; the weight differential decreased to 5%-9% by the end of the studies. Twenty-five vehicle control male mice died during weeks 48-51 from suppurative inflammation of the heart (myocarditis). At the end of the studies, the survival of male mice was as follows: vehicle control, 8/50; low dose, 28/50; high dose 31/50. Survival of female mice was lower (P<0.05) in the high dose group than in the vehicle controls (46/50; 45/50; 36/50). The primary organs affected were the forestomach (rats and mice), urinary bladder (mice), lung (mice), and liver (rats). Compound-related nonneoplastic lesions included basal cell or epithelial hyperplasia of the forestomach (rats and mice), epithelial hyperplasia of the urinary bladder (mice), and kidney hydronephrosis (mice). Neoplastic lesions associated with administration of Telone II(R) included squamous cell papillomas of the forestomach (male rats: 1/52; 1/52; 9/52; female rats: 0/52; 2/52; 3/52; female mice: 0/50; 1/50; 2/50), squamous cell carcinomas of the forestomach (male rats: 0/52; 0/52; 4/52; female mice: 0/50; 0/50; 2/50), transitional cell carcinomas of the urinary bladder (female mice: 0/50; 8/50; 21/48), alveolar/bronchiolar adenomas (female mice: 0/50; 3/50; 8/50), and neoplastic nodules of the liver (male rats: 1/52; 6/52; 7/52). Although the study in male mice was considered inadequate due to the deaths at weeks 48-51 of 25/50 vehicle control animals, 2/50 of the high dose males had transitional cell carcinomas of the urinary bladder. Furthermore, increases were seen in the incidences of alveolar/bronchiolar neoplasms of the lung (1/50; 13/50; 12/50) and of squamous cell papillomas of the forestomach (0/50; 2/50; 3/50). These findings plus the finding of nonneoplastic lesions in two of these organs (basal cell or epithelial hyperplasia of the forestomach: 0/50; 0/50; 4/50; epithelial hyperplasia of the urinary bladder: 0/50; 9/50; 18/50) suggest that Telone II(R) may have been responsible for the development of these lesions in male mice. Development of lesions of the forestomach (basal cell hyperplasia and squamous cell papilloma) was observed to be time dependent. The results of the scheduled kills supported the findings of the carcinogenesis studies. When the results from the scheduled kills at all time points were pooled with those of the 24-month carcinogenesis studies, the incidences were as follows: basal cell or epithelial hyperplasia of the forestomach-- male rats: 3/77; 13/77; 31/77; female rats: 1/75; 5/77; 35/77; squamous cell papilloma of the forestomach-- male rats: 1/77; 1/77; 13/77; female rats: 0/75; 2/77; 8/77; neoplastic nodules of the liver--male rats: 1/77; 6/76; 8/77; female rats: 6/75; 8/77; 12/77. cis- and trans-1,3- Dichloropropane are the principle components (89%) in Telone II(R), but the 1.0% epichlorohydrin, a direct-acting mutagen and carcinogen added as a stabilizer, may have influenced the development of the forestomach lesions. 1,3 Dichloropropene was mutagenic in Salmonella typhimuriumnt of the forestomach lesions. 1,3-Dichloropropene was mutagenic in Salmonella typhimurium strains TA98, TA100, and TA1535 without metabolic activation and in TA100 and TA1535 with metabolic activation by Aroclor-induced male Sprague-Dawley rat and Syrian hamster liver S9. No mutagenic response was seen in TA1537. Sex-linked recessive lethal mutations were observed in Drosophila melanogaster, and 1,3 dichloropropene did not induce reciprocal translocations in D. melanogaster. A data audit was conducted on the Telone II® 2-year carcinogenesis studies in rats and mice and the ancillary studies in rats. Except for the already known problem of survival of male vehicle control mice, no other discrepancies or problems that would compromise the validity of the findings or alter the interpretations of these studies were found. Under the conditions of these gavage studies, there was clear evidence of carcinogenicity for male F344/N rats, as indicated by Telone II®-related increased incidences of squamous cell papillomas and carcinomas of the forestomach, as well as an increased incidence of neoplastic nodules of the liver. In female F344/N rats, there was some evidence of carcinogenicity because Telone II® caused an increased incidence of squamous cell papillomas of the forestomach. The experiment in male B6C3F1 mice was considered to be an inadequate study of carcinogenicity because of reduced survival in the vehicle control group. However, there was some indication in the male mice of Telone II®-related increases of transitional cell carcinomas of the urinary bladder, squamous cell papillomas of the forestomach, and alveolar/bronchiolar adenomas and carcinomas of the lung. There was clear evidence of carcinogenicity for female B6C3F1 mice, since Telone II® caused increased incidences of transitional cell carcinomas of the urinary bladder; Telone II® also increased the incidences of alveolar/bronchiolar adenomas of the lung and of squamous cell papillomas or carcinomas of the forestomach in the female mice. Telone II®-related nonneoplastic lesions included basal cell or epithelial cell hyperplasia in the forestomach of male and female rats and male and female mice and epithelial hyperplasia of the urinary bladder in male and female mice. Synonyms: cis,trans-1,3-dichloropropene, D-D® Soil Fumigant; Telone® Soil Fumigants; Vorlex® Soil Fumigants PMID- 12748685 TI - NTP Toxicology and Carcinogenesis Studies of Propylene Oxide (CAS no. 75-56-9) in F344/N Rats and B6C3F1 Mice (Inhalation Studies). AB - Propylene oxide is a volatile, colorless liquid used as an intermediate in the production of polyether polyols, polyurethane foams, and unsaturated polyester resins and also as a fumigant for sterilizing a variety of materials ranging from plastic medical instruments to foodstuffs. In the United States, propylene oxide is registered as a fumigant for packaged dried prunes and glace fruits such as candied cherries and as an insecticidal and fungicidal fumigant for bulk quantities of cocoa, gums, and processed spices. The 2-year carcinogenesis studies of propylene oxide (greater than 99.9% pure) were conducted by exposing groups of 50 F344/N rats and 50 B6C3F1 mice of each sex to air containing propylene oxide at concentrations of 0 (chamber control), 200, or 400 ppm for 6 hours per day, 5 days per week, for 103 weeks. The survival of rats exposed to propylene oxide was comparable with that of the controls; terminal body weights were lower than those of the controls for high dose males (8%) and high dose females (6%). Survival of exposed male and female mice decreased relative to that of the controls (male: control, 42/50; low dose, 34/50; high dose, 29/50; female: 38/50; 29/50; 10/50), but the difference was significant only for animals in the high dose groups. High dose female mice had a mean terminal body weight 22% below that of the controls. The respiratory epithelium of the nasal turbinates was one of the primary tissues affected in male and female rats; exposure-related increases occurred in the incidences of suppurative inflammation, epithelial hyperplasia, and squamous metaplasia. Papillary adenomas, involving the respiratory epithelium and underlying submucosal glands of the nasal turbinates, were observed in three female rats and two male rats exposed to propylene oxide at 400 ppm. The incidence of adenomas in females was significant by the trend tests. The proportions of high dose female rats with C-cell adenomas and with C cell carcinomas of the thyroid gland were increased, but only the combined incidence of these tumors was significant (2/45; 2/35; 7/37). These tumors were not considered to be related to exposure to propylene oxide because there was no other evidence for C-cells' being a target tissue and because there was no increase in C-cell hyperplasia. The combined incidences of female rats with endometrial stromal polyps and endometrial stromal sarcomas of the uterus were significantly increased in the dosed groups (3/49; 12/50; 10/47). However, the occurrence of these lesions in the dosed groups was similar to the average (306/1,502, 20%) seen in untreated controls in NTP carcinogenesis studies, and hence this increase was not regarded as being related to exposure to propylene oxide. The respiratory epithelium of the nasal turbinates was also one of the primary tissues affected in male and female mice; exposure-related increases occurred in the incidences of inflammation, and squamous metaplasia was observed in one low dose male and two high dose female mice. One squamous cell carcinoma and one papilloma occurred in the nasal cavity of different high dose male mice, and two high dose female mice had adenocarcinomas of the nasal cavity. The endothelial cells of the submucosal vascular plexus in the nasal turbinates also appeared to be a major site affected in high dose male mice. Three high dose male and three high dose female mice had a saccular dilation (classified as angiectasis) of submucosal turbinate vessels. Further, hemangiomas were seen in the nasal cavity of 5/50 high dose male mice and 3/50 high dose female mice, and hemangiosarcomas were found in the nasal cavity of 5/50 high dose male mice and 2/50 high dose female mice. The increased incidences of hemangiomas in males and females and of hemangiosarcomas in males were statistically significant. Vascular tumors were not present in the nasal turbinates of any low dose or control mice. Under the conditions of these studies, there was some evidence of carcinogenicity for F344/N rats, as indicated by increased incidences of papillary adenomasncreased incidences of papillary adenomas of the nasal turbinates in male and female rats exposed to propylene oxide at 400 ppm. For male and female B6C3F1 mice, there was clear evidence of carcinogenicity, as indicated by increased incidences of hemangiomas or hemangiosarcomas of the nasal turbinates at 400 ppm. In the respiratory epithelium of the nasal turbinates, propylene oxide also caused suppurative inflammation, hyperplasia, and squamous metaplasia in rats and inflammation in mice. PMID- 12748686 TI - NTP Toxicology and Carcinogenesis Studies of Monuron (CAS No. 150-68-5) in F344/N Rats and B6C3F1 Mice (Feed Studies). AB - Carcinogenesis studies of monuron (greater than 99% pure), a substituted urea herbicide, were conducted by feeding diets containing 0, 750, or 1,500 ppm monuron to groups of 50 F344/N rats of each sex and 0, 5,000, or 10,000 ppm to groups of 50 B6C3F1 mice of each sex for 103 weeks. Survivors then were fed a control diet for 1 week, killed, and examined. Throughout most of the studies, mean body weights of dosed rats and mice of each sex were lower than those of the controls. Survival rates of low dose female rats and high dose male and female mice were increased relative to those of the controls. In 13-week toxicity studies, the lympho/hematopoietic system of rats and mice was the primary site affected. The lymphoid depletion found in these animals was not seen in rats or mice surviving to the end of the 104-week studies. Nonneoplastic changes associated with the long-term administration of monuron to rats included renal tubular cell cytomegaly, mainly involving the proximal convoluted tubules in male and female rats, and dose-related hepatic cytoplasmic changes in male rats. In the 104-week study, the kidneys and liver of male rats were the primary tissues affected. Long-term administration of monuron was associated with an increase in renal tubular cell adenomas (control, 0/50; low dose, 2/50; high dose, 7/50) and renal tubular cell adenocarcinomas (0/50; 1/50; 8/50). Administration of monuron to male rats was associated with increased incidences of neoplastic nodules of the liver (1/50; 6/49; 7/50) and of neoplastic nodules or carcinomas (combined) of the liver (1/50; 6/49; 9/50). Dosed male and female rats had decreased incidences of mononuclear cell leukemia; dosed male rats had lower incidences of pheochromocytomas of the adrenal glands and C-cell carcinomas of the thyroid gland; dosed female rats had reduced incidences of mammary gland fibroadenomas. In male mice, dose-related decreases occurred in the incidences of hepatocellular carcinomas (6/50; 5/49; 2/50) and hepatocellular adenomas or carcinomas (12/50; 8/49; 6/50); incidences of hepatocellular tumors in low dose female mice were reduced in dosed female mice (16/50; 8/50; 7/50). Monuron was not mutagenic in Salmonella strains TA98, TA100, TA1535, or TA1537 in the presence or absence of Aroclor 1254-induced rat liver S9. Monuron did induce chromosomal aberrations and sister chromatid exchanges in cultured Chinese hamster ovary cells. The data, documents and pathology materials from the 2-year studies of monuron have been audited. The audit findings show that the conduct of the studies is documented adequately and support the data and results given in this Technical Report. Under the conditions of these 2-year feed studies, there was clear evidence of carcinogenicity for male F344/N rats in that monuron caused increased incidences of tubular cell adenocarcinomas of the kidney, tubular cell adenomas of the kidney, and neoplastic nodules or carcinomas (combined) of the liver. Monuron induced cytomegaly of the renal tubular epithelial cells in both male and female F344/N rats. There was no evidence of carcinogenicity for female F344/N rats or for male or female B6C3F1 mice. Synonyms and Trade Names: N'-(4-chlorophenyl)-N,N dimethylurea; 1,1-dimethyl-3-(p-chlorophenyl)urea; CMU; Karmex Monuron Herbicide; Telvar PMID- 12748687 TI - NTP Toxicology and Carcinogenesis Studies of 1,2-Dichloropropane (Propylene Dichloride) (CAS No. 78-87-5) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - 1,2-Dichloropropane is a chemical intermediate widely used in the production of tetrachloroethylene and carbon tetrachloride. It is an oil and fat solvent in certain furniture finishes, dry cleaning fluids, and paint removers and has been used to fumigate grain and soil and to control peach tree borers. Approximately 77 million pounds of 1,2-dichloropropane were produced in the United States in 1980. The current 8-hour time-weighted-average concentration to which workers may be exposed to is 75 ppm. Carcinogenesis studies of 1,2-dichloropropane (<99% pure) were conducted by administering the chemical in corn oil by gavage to groups of 50 female F344/N rats and 50 male and 50 female B6C3F1 mice at doses of 125 or 250 mg/kg body weight and to groups of 50 male F344/N rats at doses of 62 or 125 mg/kg body weight. Doses were administered five times per week for 103 weeks. Vehicle control groups of 50 rats and 50 mice of each sex received an equivalent amount of corn oil by gavage on the same dosing schedule. Survival was reduced for high dose female rats (P<0.001) and for high dose female mice (P<0.05) relative to controls; 16/50 high dose female rats and 26/50 high dose female mice survived to the end of the experiment. Survival in the other groups was comparable to the control groups. In female mice, ovarian, uterine, or multiple organ infections may have contributed to the deaths of 5/11 vehicle control, 9/14 low dose, and 14/22 high dose animals that died before the end of the study. There was no evidence of an adverse effect on survival in male rats or in male mice. Mean body weights of high dose male (-14%) and high dose female ( 24%) rats were lower than those of the controls. Low dose rats and all groups of mice had mean body weights comparable to the controls. High dose female rats had increased incidences of both clear-cell changes and necrosis of the liver; but there was no increase in the incidence of liver tumors in the female rats. There was no treatment-related effects observed in the male rats, other than decreased body weights. Dose-related increases were observed for adenomas of the liver in both male (control, 7/50; low dose, 10/50; high dose 17/50) and female (1/50, 5/50, 5/50) mice. The increase in the frequency of liver carcinomas supported the evidence that there was a neoplastic response in the mouse liver for both sexes (males: 11/50, 17/50, 16/50, females: 1/50, 3/50, 4/50). Hepatocytomegaly and hepatic necrosis were increased in male mice, but not in female mice. A dose related increase in adenocarcinomas of the mammary gland was observed in female rats (control, 1/50; low dose, 2/50; high dose, 5/50) with the majority of these tumors being found at the end of the study (1/37, 3%; 2/43, 5%; 4/16, 25%). The incidence of mammary adenocarcinomas was increased when compared to the historical controls for this laboratory (3/150, 2.0%) and for all laboratories combined (11/895, 1.2%). Mammary fibroadenomas were decreased in the high dose treated female rats (15/50, 20/50, 7/50). The mutagenic activity of 1,2 dichloropropane was marginal. The compound was tested in strains TA100, TA98, TA1537, and TA1535 of Salmonella typhimurium in the presence or absence of S9 and no clearly positive response was obtained. Chromosomal aberration and sister chromatid exchange data showed that 1,2-dichloropropane caused increases in both in the absence or presence of S9. Under the conditions of these 2-year gavage studies, there was no evidence of carcinogenicity for male F344/N rats receiving 62 or 125 mg/kg. For female rats there was equivocal evidence of carcinogenicity in that 250 mg/kg 1,2-dichloropropane caused a marginally increased incidence of adenocarcinomas in the mammary gland; these borderline malignant lesions occurred concurrent with decreased survival and reduced body weight gain. There was some evidence of carcinogenicity for male and female B6C3F1 mice exposed to 1,2 dichloropropane, as indicated by increased incidences of hepatocellular neoplasms, primarily adenomas. Synonym: propylene dichloride PMID- 12748688 TI - NTP Toxicology and Carcinogenesis Studies of Chlorobenzene (CAS No. 108-90-7) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - Chlorobenzene is a colorless, volatile liquid under standard environmental conditions (vapor pressure=11.8 mm Hg at 25 degrees C, 760 mm Hg). It is used primarily as a solvent (e.g. resins, dyes, pesticides, and perfumes), a degreasing agent, and a chemical intermediate, particularly in the synthesis of nitrobenzenes. Although still considerable, estimates of the yearly production volume of chlorobenzene in the United States indicate declining use in recent years, due to the reduced demand for organochlorine pesticides utilizing chlorobenzene as an intermediate. Toxicology and carcinogenesis studies of chlorobenzene (<99% pure) were conducted by administering the test chemical in corn oil by gavage to groups of 50 male and 50 female F344/N rats and 50 female B6C3F1 mice at doses of 60 or 120 mg/kg. Groups of 50 male B6C3F1 mice received 30 or 60 mg/kg. Chlorobenzene was administered five times per week for 103 weeks. Groups of 50 rats and 50 mice of each sex received corn oil by gavage on the same schedule and served as vehicle controls, and additional groups of 50 rats and 50 mice of each sex served as untreated controls. The chlorobenzene doses were chosen on the basis of 90-day studies, in which doses 2-fold or greater in excess of the doses used in the 2-year study caused death, hepatocellular necrosis, renal tubular injury, thymic necrosis, or lymphoid or myeloid depletion of bone marrow, spleen or thymus. Mean body weights of dosed rats and mice were essentially the same or greater than those of the controls during the 2-year studies. Survivals of low dose male rats, dosed female rats, dosed male mice, and dosed female mice were not adversely affected by administration of chlorobenzene. Survival of high dose male rats in the 2-year study was significantly (P=0.033) lower than that of the vehicle controls. No chlorobenzene-induced toxic lesions responsible for this reduction in survival were observed. Based on the prechronic results and on the above data, the doses used in the 2-year study were considered to be adequate for carcinogenicity testing. Male rats dosed with chlorobenzene exhibited a significant (P<0.05) increase in the incidence of animals with neoplastic nodules of the liver (overall incidences: untreated control, 4/50 (8%); vehicle control, 2/50 (4%); low dose, 4/49 (8%); high dose, 8/49 (16)). Increased incidences of hepatocellular carcinomas in male rats or of neoplastic nodules or hepatocellular carcinomas in female rats were not observed. No increased tumor incidences were observed in female rats or in male or female mice. Under the conditions of these studies, chlorobenzene administration increased the occurrence of neoplastic nodules of the liver in high dose (120 mg/kg/day) male F344/N rats, providing some but not clear evidence of carcinogenicity of chlorobenzene in male rats. Carcinogenic effects of chlorobenzene were not observed in female F344/N rats or in male or female B6C3F1 mice. Levels of Evidence of Carcinogenicity: Male Rats: Equivocal Female Rats: Negative Male Mice: Negative Female Mice: Negative Synonyms: monochlorobenzene; chlorobenzol; phenyl chloride; benzene monochloride PMID- 12748689 TI - NTP Carcinogenesis Studies of Ethyl Acrylate (CAS No. 140-88-5) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - Ethyl acrylate is a monomer used to produce polymers and copolymers for use in latex paints, textiles, paper coatings, fabric finishes, dirt release agents, and specialty plastics. In 1980, 268 million pounds of ethyl acrylate were produced in the United States of which 209 million pounds were used by the producers and 59 million pounds sold. Carcinogenesis studies of ethyl acrylate were conducted by administering this test chemical in corn oil by gavage to groups of 50 male and 50 female F344/N rats and B6C3F1 mice at doses of 100 or 200 mg/kg. Ethyl acrylate was administered five times per week for 103 weeks. Groups of 50 rats and 50 mice of each sex received corn oil by gavage on the same schedule and served as vehicle controls. Survival of dosed male and female rats and mice was comparable with that of the corresponding vehicle controls. There was no evidence of systemic toxicity in the prechronic or in the 2-year studies. Compound-related increased incidences of hyperkeratosis, inflammation, and hyperplasia of the forestomach were observed in rats and mice in the prechronic as well as 2-year studies. In the 2-year studies, squamous cell papillomas and squamous cell carcinomas of the forestomach occurred at the site of chemical deposition with significant positive trends and increased incidences in dosed groups versus vehicle controls for both sexes of rats and mice. Nonneoplastic and neoplastic forestomach lesion frequencies were related to the concentration of ethyl acrylate in dosing solutions used. Significant negative trends for several common rodent tumors were found in treated animals in the 2-year studies. Under the conditions of these studies, ethyl acrylate was carcinogenic for the forestomach of F344/N rats and B6C3F1 mice, causing squamous cell carcinomas in male rats and male mice, squamous cell papillomas in male and female rats and male mice, and squamous cell papillomas or carcinomas (combined) in male and female rats and mice. Evidence for carcinogenicity was greater in males than in females. Ethyl acrylate also caused irritation of the forestomach mucosa in male and female rats and mice. Levels of Evidence of Carcinogenicity: Male Rats: Positive Female Rats: Positive Male Mice: Positive Female Mice: Positive Synonym: 2-propanoic acid, ethyl ester PMID- 12748690 TI - NTP Toxicology and Carcinogenesis Studies of Diglycidyl Resorcinol Ether (Technical Grade) (CAS No. 101-90-6) In F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - Diglycidyl resorcinol ether (DGRE), a pale, yellow, translucent, amorphous solid at room temperature, is used as a liquid spray epoxy resin, as a diluent in the production of other epoxy resins used in electrical, tooling, adhesive, and laminating applications, and as a curing agent for polysulfide rubber. Approximately 3,000 workers are exposed to DGRE. The quantity of DGRE produced in the United States is not known. Toxicology and carcinogenesis studies of technical grade diglycidyl resorcinol ether (81% pure) were conducted by administering the chemical in corn oil by gavage to groups of 50 male and 50 female F344/N rats at doses of 25 or 50 mg/kg and to groups of 50 male and female B6C3F1 mice at doses of 50 or 100 mg/kg. A supplemental study of similar design in male and female rats (0 or 12 mg/kg) was started approximately 12 months later because of high mortality in the 50 mg/kg dose groups. Doses were administered five times per week for 103 weeks. Groups of 50 rats and 50 mice of each sex received corn oil by gavage on the same dosing schedule and served as vehicle controls. Throughout most of the primary study, mean body weights of high dose male and female rats and female mice were lower than those of the corresponding vehicle controls. In the supplemental study, body weights of both sexes of the dosed rats were unaffected by administration of DGRE. Survival of dosed rats of each sex in the primary study was dose related and was shorter (P<0.001) than that of the vehicle controls. No high dose male rats and only 1/50 high dose female rats lived to the end of the study. Bronchopneumonia was the most frequent cause of early death among the rats and may have resulted from the animals' aspiration of corn oil containing diglycidyl resorcinol ether. Survival of the dosed male rats in the supplemental study was reduced (P<0.005) when compared to controls. There was no significant difference in survival between dosed and control female rats in the supplemental study. Survival of dosed and control mice was comparable but poorer in females, with 20/50 (40%) of the controls, 13/50 (26%) of the low dose, and 10/50 (20%) of the high dose groups alive at the end of 2 years. These early deaths were due to suppurative and necrotizing inflammation of the reproductive tract, possibly caused by a Klebsiella sp. infection. The incidences of rats and mice with hyperkeratosis and hyperplasia of the forestomach were compound related. For rats and mice of each sex, incidences of animals with squamous cell papillomas, squamous cell carcinomas, or both occurred with statistically significant positive trends and the incidences observed in other organs in dosed groups relative to the controls. An audit of the experimental data was conducted for the 2-year studies of diglycidyl resorcinol ether. No data discrepancies were found that influenced the final interpretations. Under the conditions of these 2-year gavage studies, technical grade diglycidyl resorcinol ether caused hyperkeratosis and hyperplasia of the forestomach in rats and mice. DGRE was carcinogenic for male and female F344/N rats and for male and female B6C3F1 mice, causing both benign and malignant neoplasms of the forestomach. Levels of Evidence of Carcinogenicity: Male Rats: Positive Female Rats: Positive Male Mice: Positive Female Mice: Positive Synonym: DGRE PMID- 12748691 TI - NTP Toxicology and Carcinogenesis Studies of 1,2-Dichlorobenzene (o Dichlorobenzene) (CAS No. 95-50-1) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - The major use of 1,2-dichlorobenzene is as an intermediate in the synthesis of several organic compounds (e.g. 3,4-dichloroaniline) and in the syntheses of the herbicides propanil, diuron, and neburon. It is used as an industrial solvent for toluene diisocyanate, an additive to degreasing agents, a heat exchange medium, a deodorant for garbage and sewage, an ingredient in paint removers, an engine cleaner and de-inking solvent, and a solvent and intermediate in dye manufacture). 1,2-Dichlorobenzene is also used as an insecticide and a fumigant to control peach tree borers, bark beetles, grubs, and termites and to kill mites and other insects in poultry houses and animal sleeping quarters. Because of its properties as both an insecticide and a solvent, 1,2-dichlorobenzene has been used in low-pressure aerosol formulations of insecticides. Approximately 49 million pounds of 1,2-dichlorobenzene were produced in the United States in 1980. First reported commercial production began in 1921. In 13-week studies using F344/N rats and B6C3F1 mice, 500 mg/kg of 1,2-dichlorobenzene (<99% pure) decreased survival in male and female mice and female rats when administered in corn oil by gavage five times per week. At this dose, 1,2-dichlorobenzene produced centrolobular necrosis of the liver, hepatocellular degeneration, and depletion of lymphocytes in the thymus and spleen of both sexes of rats and mice. At a dose of 250 mg/kg, necrosis of individual hepatocytes was observed in both sexes of rats and in male mice. Minimal hepatocellular necrosis was observed in a few rats at a dose of 125 mg/kg, but no hepatic alterations were observed in mice at this dose. Renal tubular degeneration was observed in male rats at 500 mg/kg, and multifocal mineralization of the myocardial fibers of the heart and skeletal muscle were seen in mice. The only hematologic changes considered notable were slight decreases in hemoglobin and hematocrit in the 500 mg/kg male and female rats and in red blood cell counts in the 500 mg/kg male rats; no other marked hematological changes were observed in either species. Two-year toxicology and carcinogenesis studies of 1,2-dichlorobenzene were conducted by administering the test chemical in corn oil gavage five times per week for 103 weeks to groups of 50 male and 50 female F344/N rats and B6C3F1 mice at doses of 60 and 120 mg/kg. Groups of 50 rats and 50 mice of each sex received corn oil by gavage on the same schedule and served as vehicle controls. Survivals of female rats, male mice, and female mice were comparable to those of the corresponding vehicle controls in the 2-year study, but survival of high dose male rats was (P<0.001) shorter than that of the vehicle controls. In this group there were three accidental deaths and five deaths probably due to the gavage process; in addition aspiration of 1,2 dichlorobenzene in corn oil into the lungs may have been a contributing factor to the deaths of 12 high dose male rats. The 120 mg/kg dose level of 1,2 dichlorobenzene did not affect body weight in rats or mice of either sex or survival of mice or female rats. An increase in tubular regeneration in the kidney of high dose male mice was observed in the 2-year study (control, 8/48, 17%; low dose, 12/50, 24%; high dose, 17/49, 35%). No other compound-related nonneoplastic histological lesions were noted in the 2-year study. The incidence of pheochromocytoma of the adrenal gland in low dose male rats was elevated (P<0.05, life table test) relative to controls (9/50, 16/50, 6/49). However, the incidence in the high dose group was lower than that of the controls and the dose response trend was not statistically significant. Therefore, the increase in pheochromocytoma in the low dose male rats is not regarded as related to administration of 1,2-dichlorobenzene. A dose-related increase (P<0.05) in malignant histiocytic lymphoma was observed in male mice (control, 0/50, 0%; low dose, 1/50, 2%; high dose, 4/50, 8%) and in female mice (0/49, 0%; 0/50, 0%; 3/49, 6%); however, comparisons of the numbers of animals with all typnt;; 3/49, 6%); however, comparisons of the numbers of animals with all types of lymphomas is considered to be a more appropriate comparison. 1,2-Dichlorobenzene did not increase the incidence of all types of lymphomas (combined) in male mice (8/50, 16%; 2/50, 4%; 4/50, 8%) or female mice (11/49, 22%; 11/50, 22%; 13/49, 27%). Therefore, the increase in histiocytic lymphomas was discounted. Under the conditions of these two-year gavage studies, there was no evidence of carcinogenicity of 1,2-dichlorobenzene for male or female F344/N rats or B6C3F1 mice receiving 60 or 120 mg/kg per day. Levels of Evidence of Carcinogenicity: Male Rats: Negative Female Rats: Negative Male Mice: Negative Female Mice: Negative Synonym: o-dichlorobenzene PMID- 12748692 TI - Carcinogenesis Studies of Allyl Isovalerate (CAS No. 2835-39-4) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - Allyl isovalerate, a synthetic fragrance and flavoring ingredient in use since the 1950's, may be found in various products at the following concentrations: soap, 30 ppm; detergent, 3 ppm; creams, 15 ppm; perfume, 50 ppm; nonalcoholic beverages, 9 ppm; ice cream, 18 ppm; candy, 22 ppm; baked goods, 15-48 ppm; and gelatins and puddings, 1 ppm. A colorless liquid with an apple-like odor and taste, allyl isovalerate is approved by the U.S. Food and Drug Administration for use in foods. Specific production figures are not available, but U.S. production in 1980 exceeded 1,000 pounds. Carcinogenesis studies of allyl isovalerate (96% pure) were conducted by administering the test chemical in corn oil gavage to groups of 50 male and 50 female F344/N rats and to groups of 50 male and 50 female B6C3F1 mice at doses of 31 or 62 mg/kg. The doses selected were based on the chemically-induced toxic effects and depressed weight gains obtained from the 13-week studies. Doses were administered five times per week for 103 weeks. Groups of 50 rats and 50 mice of each sex received corn oil by gavage on the same dosing schedule and served as vehicle controls. Survival and mean body weight gain of rats of each sex and male mice were not adversely affected by the administration of allyl isovalerate. The significantly lower survival (P=0.001) and the lower mean body weight of low-dose female as compared with controls are likely consequences of the high incidence of a genital tract infection in the low dose females. This infection was probably responsible for the deaths of 11/19 control, 22/33 low-dose, and 13/25 high-dose female mice that died before the end of the study. Squamous cell papillomas and epithelial hyperplasia of the nonglandular stomach were observed in dosed male mice in the 2-year studies (squamous cell papillomas: 0/50, 1/50, 2%, 3/48, 6%; epithelial hyperplasia: 1/50, 2%, 1/50, 2%, 7/48, 15%). The papillomas occurred with a significant positive trend (P<0.05). The incidence of high-dose male mice with squamous cell papillomas of the nonglandular stomach was also higher (P<0.01) than the historical rate for vehicle control male B6C3F1 mice in the Bioassay Program (5/881, 0.6%). Forestomach lesions were also observed in female mice: squamous cell papillomas (1/50, 0/50, 2/50) and epithelial hyperplasia of the nonglandular stomach (0/50, 2/50, 3/50). Pancreatic acinar-cell adenomas occurred at higher incidences in the dosed male rats than in the controls (control, 1/50, 2%; low dose, 4/50, 8%; high-dose, 2/50, 4%). Pancreatic acinar-cell tumors were not observed in female rats. Preputial gland adenomas were observed in increased incidence in low-dose male rats (0/50, 4/50, 8%; P<0.05, 1/50, 2%). Mononuclear cell leukemias in rats and lymphomas in mice occurred with increased incidences. This consistent dose-response increase among both rats and mice indicates that allyl isovalerate adversely affects the hematopoietic system. Cholangiofibrosis, nodular regeneration, cirrhosis, focal necrosis, fatty metamorphosis, and cytoplasmic vacuolization were observed at increased incidences in the livers of high-dose male and female rats in the 2-year study. No compound-related nonneoplastic lesions were observed in the mice of either sex. Liver neoplasms were not increased in either dosed rats or mice of either sex. Significant (P<0.05) decreases in tumor incidences were observed in male mice for hepatocellular carcinomas (18/50, 6/50, 9/50), for alveolar/bronchiolar adenomas or carcinomas (13/50, 6/50, 5/49), and for follicular-cell adenomas of the thyroid gland (5/47, 0/46, 1/49). Allyl isovalerate was not mutagenic for Salmonella typhimurium (tester strains TA 98, 100, 1535, and 1537) with or without metabolic activation. Under the conditions of these studies, allyl isovalerate was carcinogenic for F344/N rats and B6C3F1 mice, causing increased incidences of hematopoietic system neoplasms (mononuclear-cell leukemia in male rats and lymphoma in female mice). Levels of Evidence of Carcinogenicity: Male Rats: Positive Female Rats: Negativen female mice). Levels of Evidence of Carcinogenicity: Male Rats: Positive Female Rats: Negative Male Mice: Negative Female Mice: Positive PMID- 12748693 TI - NTP Carcinogenesis Studies of Food Grade Geranyl Acetate (71% Geranyl Acetate, 29% Citronellyl Acetate) (CAS No. 105-87-3) in F344/N Rats and B6C3F1 Mice (Gavage Study). AB - Geranyl acetate (3,7-dimethyl-2,6-octadiene-1-ol acetate) is a colorless liquid prepared by fractional distillation of selected essential oils or by acetylation of geraniol. It is a natural constituent of more than 60 essential oils, including Ceylon citronella, palmarosa, lemon grass, petit grain, neroli bigarade, geranium, coriander, carrot, and sassafras. Geranyl acetate is used primarily as a component of perfumes for creams and soaps and as a flavoring ingredient. On the U.S. Food and Drug Administration's list of substances "generally recognized as safe," the Food Chemicals Codex (1972) specifies that geranyl acetate must contain at least 90% total esters. Carcinogenesis studies of food-grade geranyl acetate (containing approximately 29% citronellyl acetate) were conducted by administering the test chemical in corn oil by gavage to groups of 50 male and 50 female F344/N rats at doses of 1,000 or 2,000 mg/kg body weight and to groups of 50 male and 50 female B6C3F1 mice at doses of 500 or 1,000 mg/kg. Doses were administered five times per week for 103 weeks. Groups of 50 rats and 50 mice of each sex received corn oil by gavage on the same dosing schedule and served as vehicle controls. The cumulative toxicity of geranyl acetate in the 2-year study was indicated by the significantly shorter survival of high dose male rats (control, 34/50; low dose, 29/50; high dose, 18/50) and of high dose male mice (control, 31/50; low dose, 32/50; high dose, 0/50) and of dosed female mice (38/50; 15/50; 0/50) when compared with controls. Throughout most of the 2-year study, mean body weights of high dose rats and mice of each sex were lower than those of the controls. The occurrence of retinopathy or cataracts in the high dose male rats and low dose female rats as compared with the controls does not appear to be related to the administration of geranyl acetate but rather the proximity of the rats to fluorescent light. The incidence of retinopathy or cataracts (combined) was: males: control, 0/50, 0%; low dose, 1/50, 2%; high dose, 11/50, 22%; females: control, 1/50, 2%; low dose, 13/50, 26%; high dose, 2/50, 4%. Kidney tubular cell adenomas, an uncommon tumor type, were found in 2/50 (4%) low dose male rats. The historical incidence of male corn oil gavage control F344/N rats with kidney tumors is 1/250 (0.4%) at this laboratory and 4/998 (0.4%) in the program. Squamous cell papillomas in the skin were increased marginally in low dose male rats (control, 0/50; low dose, 4/50, 8%; high dose, 1/50, 2%). In addition, one low dose male rat had a squamous cell carcinoma of the skin. The incidence of low dose male rats with either squamous cell papillomas or carcinomas was greater (P<0.05) in comparison with the controls. The historical incidence of squamous cell papillomas or carcinomas (combined) in gavage control male F344/N rats is 3.6% (9/250) at this laboratory and 2.5% (25/999) throughout the program. The incidence of all epidermal tumors was not significantly elevated in dosed male rats relative to controls (control, 3/50, 6%; low dose, 6/50, 12%; high dose, 1/50, 2%). All high dose (1,000 mg/kg) male and female mice were dead by week 91 as a result of accidentally being administered 2,800 mg/kg for 3 days during week 91; survival of low dose and control male mice was comparable. Survival of high dose male and dosed female mice may have been inadequate for the detection of late-appearing tumors. No evidence of any carcinogenic effect was found in either low or high dose mice of either sex. An infection of the genital tract was probably responsible for the deaths of 14/22 control and 8/32 low dose female mice before the end of the study. Cytoplasmic vacuolization was increased in the liver and in the kidney of male and female mice and was considered to be compound related (liver-- male: control, 1/50, 2%; low dose, 7/50, 14%; high dose, 47/50, 94%; female: 1/50, 2%; 27/50, 54%; 46/50, 92%; kidney or kidney tubule--male: 0/50; 0/50; 41/50, 82%; female: 0/50; 24/49, 49%; 37/50, 74%). Under the conditions of these studies, geranyl 4%; 46/50, 92%; kidney or kidney tubule--male: 0/50; 0/50; 41/50, 82%; female: 0/50; 24/49, 49%; 37/50, 74%). Under the conditions of these studies, geranyl acetate was not carcinogenic for F344/N rats or B6C3F1 mice of either sex; however, the reduced survival observed in high dose male rats, high dose male mice, and high and low dose female mice lowered the sensitivity of these studies for detecting neoplastic responses in these groups. In male rats the marginal increases of squamous cell papillomas of the skin and tubular cell adenomas of the kidney may have been related to administration of geranyl acetate. Levels of Evidence of Carcinogenicity: Male Rats: Negative Female Rats: Negative Male Mice: Negative Female Mice: Negative PMID- 12748694 TI - NTP Toxicology and Carcinogenesis Studies of Commercial Grade 2,4 (80%)- and 2,6 (20%)- Toluene Diisocyanate (CAS No. 26471-62-5) in F344/N Rats and B6C3F1 Mice (Gavage Studies). AB - Toluene diisocyanate (TDI) is commercially produced as an approximate 80:20 mixture of the 2,4- and 2,6-isomers. In 1980, 580,000 pounds of this chemical were produced in the United States, primarily for use in the manufacture of flexible polyurethane foams. These foam elastomers are found in furniture and automobile cushions, carpet underlays, pillow filling, mattresses, insulation, shoes, purses, and toys. TDI is also used to produce polyurethane coatings for lacquers and wood finishes. Groups of 50 female F344/N rats and 50 B6C3F1 mice were administered commercial grade toluene diisocyanate (80% 2,4- and 20% 2,6-) in corn oil by gavage at doses of 60 or 120 mg/kg body weight, 5 days per week for 105 or 106 weeks. Groups of 50 male F344/N rats received 30 or 60 mg/kg and groups of 50 male B6C3F1 mice received 120 or 240 mg/kg on the same schedule. Dosage analyses of toluene diisocyanate indicated that the chemical had reacted in the corn oil vehicle, resulting in actual gavage concentrations 77% to 90% of theoretical values. Groups of 50 rats and 50 mice of each sex received corn oil only and served as vehicle controls. Survival in all groups of dosed rats in the 2-year studies were shorter (P5) LDL:HDL ratios and the development of lens opacities (p<0.0003). The risk of falling into the cataract subgroup if the individual's LDL:HDL ratio exceeded 5 was 2.35 (95% CI = 1.09-5.04; p = 0.014). CONCLUSION: This study strongly suggests that an association exists between low levels of HDL cholesterol and high LDL:HDL ratios on one hand and the development of adult lens opacification on the other. PMID- 12748743 TI - Long-term follow-up of single-lead VDD pacing. AB - Long-term outcomes of single-lead VDD pacing were studied retrospectively and partly prospectively. Records were analysed of 81 patients out of 133 in whom a single-lead VDD pacemaker was implanted between January 1993 and December 1997 and who attended a follow-up clinic more than two years after the implant. Forty eight of them attended a prospective follow-up 54 +/- 15 months after the implant. Sinus rhythm was present in 91.5% of the patients and atrial fibrillation in the remaining 8.5%. A-V synchronous pacing was documented in 91.9 to 94.9% at different follow-up periods; however, an intermittent asynchronous ventricular (VVI) pacing of more than 10% occurred intermittently in 19.1% of the patients. Chronic sensed P-wave amplitude was significantly lower than the implant P-wave amplitude (by 70%) and did not correlate with the implant amplitude. Postural changes (supine, sitting, standing, with normal breathing and during deep inspiration) did not have a significant impact on sensed P-wave amplitude more than four years after the implant. Rate histograms were remarkably stable over the years, with dominant heart rate 70 to 79 beats per minute observed for 25 to 30% of the monitored periods. Single-lead VDD pacing was found to be a reliable method of long-term physiological pacing in patients with heart block who returned for follow-up. Routine testing more than four years after the implant does not require postural manoeuvres. PMID- 12748745 TI - Dietary markers of hypertension associated with pulse pressure and arterial compliance in black South African children: the THUSA Bana Study. AB - AIM: The aim of the study was to determine which dietary factors contribute to the impairment of arterial compliance, stroke volume, total peripheral resistance and pulse pressure, and could thereby be identified as risk markers in the development of hypertension in black children. METHODS: Children aged 10 to 15 years were recruited from 30 schools in the North West Province over two years (2000 to 2001). These children comprised 321 black males and 373 females from rural to urbanised communities and 40 male and 79 females subjects with identified high-normal to hypertensive blood pressure. Blood pressure was measured by means of a Finapres apparatus. Through analysis with the Fast Modelflo software program, systemic arterial compliance, pulse pressure, total peripheral resistance and stroke volume were obtained. A 24-hour dietary recall questionnaire and weight and height measurements were taken. RESULTS: In a stepwise regression analysis the following nutrient were significantly associated (p < or = 0.05) with cardiovascular parmeters of hypertensive subjects: protein, carbohydrates, total fat, polyunsaturated fat, mono-unsaturated fat, saturated fat, fibre, vitamin A, nicotinic acid, biotin, vitamin B(12), ascorbic acid, vitamin E, magnesium, manganese, phoshorus and iron. No significant dietary markers were indicated for the normotensive groups. Dietary intakes of most of these nutrients were below the dietary reference intakes for all groups. CONCLUSIONS: The results indicate strong associations of protein, polyunsaturated fats, fibre, vitamin A, vitamin C, vitamin E, nicotinic acid, vitamin B(12), biotin and phosphorus with the rate of hypertension in black South African children. PMID- 12748744 TI - Mechanisms of opioid delta (delta) and kappa ( kappa) receptors' cardioprotection in ischaemic preconditioning in a rat model of myocardial infarction. AB - It has been demonstrated that brief periods of coronary artery occlusion before a prolonged period of sustained occlusion paradoxically protect the myocardium against infarction. The mechanisms involved in this phenomenon, termed "ischaemic preconditioning" (IPC) are still not clear, although it has been established that opioid receptors are involved. The aim of this study was to probe some of the plausible mechanisms involved in the phenomenon by using an in vivo model of myocardial infarction in intact rat, a model that allows electro-cardiographic and enzymatic in addition to morphometric evaluation of the development of 24 hour myocardial infarction. Selective opioid delta-receptor agonist (DADLE) and antagonist (natrindole), and opioid kappa-receptor agonist (U-50488H) and antagonist (nor-BNI) were used. To clarify some of the mechanisms of IPC, we used selective inhibitors of the anticipated cellular systems involved. Pertussis toxin (inhibitor of adenylate cyclase G(I/o) protein), glibenclamide (inhibitor of K(ATP ) channel) and chelerythrine (inhibitor of PKC) were used. Results obtained showed that: Both opioid delta- and kappa-receptors were involved in the beneficial effect of IPC, although we were unable to differentiate between opioid receptor subtypes (delta1, delta2 and kappa1, kappa2). Opioid delta- and kappa receptors displayed different effects in IPC. After 30 minutes of left coronary occlusion and 2-hour reperfusion, opioid delta-receptor agonist DADLE significantly decreased (p < 0.05) the infarct size (by 66%--from % IS/AAR 59.80 in the control, untreated infracted rats to % 20.40), without a significant effect (p > 0.05) on the occurrence of early arrhythmias. Opioid kappa-receptor agonist U-50488H produced mainly antiarrhythmic effects. It decreased % IS/AAR by 44%, reduced the occurrence of early arrhythmias by 77%, and decreased ventricular ectopic beats by 80%. Both opioid delta- and kappa-receptor agonists significantly reduced (p < 0.05 ) early (2-hour) mortality by 22% and 19% respectively. The above opioid delta- and kappa-receptor cardiac effects were abolished by the use of respective specific opioid delta- and kappa-receptor antagonists. The beneficial effects of opioid delta- and kappa-receptor agonists persisted for at least 24 hours post-infarction. It is most likely that both opioid delta- and kappa-receptors act via common cellular mechanisms involving: activation of ATP-sensitive (sarcolemmal) K+ channel via G(I/o) proteins (based on the results of our experiments with K(ATP) channel antagonist, glibenclamide); phosphatidylinositol pathway via activation of protein kinase C (judging from the results of our experiments with the inhibitor of PKC, chelerythrine); and the recently proposed "cross talk" between beta (1)-adrenergic and opioid receptors in cardiac myocytes (involving inhibition of adenylate cyclase by G(I/o) proteins). Exploring the possibility of this signaling pathway will be the next step in our experimental studies. PMID- 12748746 TI - "Low pressure" left ventricular tamponade in a patient with rheumatic mitral stenosis and HIV-related acute pericarditis. AB - A case report of isolated left ventricular tamponade in a patient with rheumatic mitral stenosis and effusive pericarditis is presented. The haemodynamics and management of this under-diagnosed pathology is discussed. PMID- 12748747 TI - Long-term follow-up of single-lead VDD pacing. PMID- 12748749 TI - The determination of water in crude oil and transformer oil reference materials. AB - The measurement of the amount of water in oils is of significant economic importance to the industrial community, particularly to the electric power and crude oil industries. The amount of water in transformer oils is critical to their normal function and the amount of water in crude oils affects the cost of the crude oil at the well head, the pipeline, and the refinery. Water in oil Certified Reference Materials (CRM) are essential for the accurate calibration of instruments that are used by these industries. Three NIST Standard Reference Materials (SRMs) have been prepared for this purpose. The water in these oils has been measured by both coulometric and volumetric Karl Fischer methods. The compounds (such as sulfur compounds) that interfere with the Karl Fischer reaction (interfering substances) and inflate the values for water by also reacting with iodine have been measured coulometrically. The measured water content of Reference Material (RM) 8506a Transformer Oil is 12.1+/-1.9 mg kg(-1) (plus an additional 6.2+/-0.9 mg kg(-1) of interfering substances). The measured water content of SRM 2722 Sweet Crude Oil, is 99+/-6 mg kg(-1) (plus an additional 5+/-2 mg kg(-1) of interfering substances). The measured water content of SRM 2721 Sour Crude Oil, is 134+/-18 mg kg(-1) plus an additional 807+/-43 mg kg(-1) of interfering substances. Interlaboratory studies conducted with these oil samples (using SRM 2890, water saturated 1-octanol, as a calibrant) are reported. Some of the possible sources of bias in these measurements were identified, These include: improperly calibrated instruments, inability to measure the calibrant accurately, Karl Fischer reagent selection, and volatilization of the interfering substances in SRM 2721. PMID- 12748750 TI - Introduction of high carbon content solvents into inductively coupled plasma mass spectrometry by a direct injection high efficiency nebuliser. AB - A preliminary investigation on the introduction of high carbon content solvents into inductively coupled plasma mass spectrometry (ICP-MS) by a direct injection high efficiency nebuliser (DIHEN) is presented. Ethanol, hexane, toluene and a natural gas condensate were introduced using a flow injection system. The performance for determinations of total concentrations of mercury present in organic solvents as different species was evaluated. The most critical operating parameters were the nebuliser gas flow rate and amount of oxygen added to the plasma. For the DIHEN a nebuliser gas flow rate of 0.3 L min(-1) and 50 mL min( 1) of oxygen added to the plasma auxiliary gas flow gave stable conditions and high analyte sensitivity. Species recoveries for HgCl(2), CH(3)HgCl, (CH(3))(2)Hg and Hg(0) in hexane were 99+/-4, 101+/-4, 95+/-4 and 104+/-7%, respectively. Detection limit for mercury in hexane was 85 pg mL(-1) based on 3 sigma of a (201)Hg-spiked blank. A gradual deposition was observed at the nebuliser tip, partly blocking the gas annulus. The rate of deposition seemed to be related to the amount of carbon introduced through the nebuliser. With the optimised conditions used in this work, the nebuliser could be used for approximately 50 hexane samples before cleaning was necessary. PMID- 12748751 TI - The role of organic colloids in herbicide transfer to rivers: a quantitative study of triazine and phenylurea interactions with colloids. AB - For moderately hydrophobic compounds such as most pesticides adsorption on colloids (<0.2 microm) may play a key role in pesticide mobility as well as in their degradation by chemical and microbiological processes. However, until now, pesticide-organic colloid interactions are poorly understood. Quantitative data for sorption equilibria on colloids of two series of herbicides including triazines (atrazine, simazine, terbutylazine, prometryne, desethylatrazine, and desisopropylatrazine) and phenylureas (isoproturon, linuron, neburon, and diuron) sampled in the Seine river (urban zone) and the Marne river (agricultural zone) are presented. Partition coefficient of herbicides on colloids (K(com)), were evaluated by solid-phase extraction coupled with high-performance liquid chromatography-UV diode-array detection (SPE-HPLC-UV/DAD). In the case of triazines a satisfactory log-log correlation was found between K(com) and octanol water coefficient (K(ow)) values. Phenylureas did not obey this correlation, with K(com) values being about two times higher than those of triazines. The existence of two distinct types of adsorption behaviour on colloids partly explains the different occurrence of triazines and phenylureas in surface waters. PMID- 12748752 TI - Three-phase plasma arc atomic-emission spectrometric analysis of environmental samples using an ultrasonic nebulizer. AB - Combination of an ultrasonic nebulizer and plasma excitation sources for spectrochemical analysis offers desirable features of low detection limits, high sample throughput, wide dynamic range of operation, acceptable precision and accuracy, and simultaneous quantitative analytical capabilities. Moreover, the ultrasonic nebulizer does not require sample preconcentration. Recently we have developed a three-phase plasma arc (TPPA) for atomic emission spectrochemical analysis. In the present work, to increase the analytical utility of the three phase plasma system, an ultrasonic nebulizer was used for sample introduction. The effects of the argon gas flow rate, current, excitation temperature have been studied. The analytical calibration curves are obtained for Ca, Cr, Fe, Mg and Mn, and detection limits have been calculated. The present technique is used to determine the concentration of the elements Ca, Cr, Fe, Mg and Mn in airborne samples. PMID- 12748753 TI - Characterization of carbohydrates in mucilage samples from the northern Adriatic Sea. AB - Carbohydrate contents in seawater, mucilage, and mucilage interstitial waters were analyzed during episodes of mucilage formation in the summers of 2000 and 2001 in the northern Adriatic Sea off Pesaro and in the Gulf of Trieste using 3 methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH) and 2,4,6-tripyridyl-s triazine (TPTZ) assays. The significant presence of polysaccharides in seawater in the presence of mucilage has an important impact on the agglomeration processes forming gelatinous material (macrogels). Characterization of oligosaccharides in the water-soluble fraction of mucilage using HPLC/RI revealed maltose and pentaose as the main components. PMID- 12748754 TI - Development of an EnFET for the detection of organophosphorous and carbamate insecticides. AB - A biosensor for the detection of insecticides based on an ion-sensitive field effect transistor (ISFET) was developed. The resulting device combines the simplicity of potentiometric sensors and the use of associated electronic systems as powerful tools for the acquisition and the processing of data. The enzyme acetylcholinesterase (AChE) was entrapped in a membrane placed on the gate of the ISFET forming an enzyme field-effect transistor (EnFET). The biosensor is applied to the determination of pesticides in spiked real samples. Organophosphorous and carbamate insecticides were measured with a detection limit of 10(-8) mol L(-1). The measurement is based on the production of hydrogen ions due to the hydrolysis of acetylthiocholine by the enzyme. The resulting local pH change is picked up by the underlying pH-sensitive ISFET and transduced as potential variations. The preparation of the membrane is simple and reproducible. The analysis in spiked real samples was performed in tap water and showed detection limits comparable to those obtained by other researchers. PMID- 12748755 TI - Chemometric investigation of the efficiency of different TiO(2)-based catalysts as principal components of TOC photochemical sensors under development. AB - There is an increasing demand for measurement of organic carbon in solutions both in industry and in environmental research for the purpose of continuous water quality monitoring. Practically all the methods used are based on a catalytic system in which metal oxides play a major role. The development of a new TOC/DOC measurement system, in our work, uses a mixed anatase-rutile form of TiO(2 )as catalyst of the mineralisation process, and direct measurement of the CO(2) produced, by a gaseous diffusion electrode. The entire research is based on heterogeneous catalysis using an immobilised catalyst, which can offer considerable advantages over other methods of catalysis. Four different catalytic systems were analysed. Three involved thermal immobilisation of TiO(2) on glass supports (glass spheres and glassy particulates) or on an Al metal grid. One further system consisted of direct oxidation of a Ti grid at high temperatures. The system was illuminated using a 350 nm UV source (350 mW cm(-2)) contained in a home-made measurement cell. Five molecules-malic acid, pentachlorophenol, sodium dodecylsulfate, hydroquinone, and citric acid-which were deemed to be representative and had been investigated in several previous studies, were used as photodegradation targets. However, it was not possible to obtain a direct expression of "catalytic efficiency" simply by reading the data obtained, nor any indirect molecular "recalcitrancy" scale. Chemometric analysis, by principal components analysis, allows the five used catalytic systems to be easily compared each other and a single PC component is able to perform classification. PMID- 12748757 TI - Post-pubertal emergence of disrupted latent inhibition following prenatal immune activation. AB - RATIONALE: There is evidence pointing to an association between prenatal exposure to infection and increased liability to schizophrenia, and it has been suggested that the maternal immune response, in particular, the release of pro-inflammatory cytokines, may interfere with normal fetal brain development. Impaired capacity to ignore irrelevant stimuli is considered one of the central deficits in schizophrenia, and is manifested, among others, in disrupted latent inhibition (LI). OBJECTIVES: To test the effects of prenatal immune activation on LI in juvenile and adult offspring. METHODS: Pregnant rats were injected with the synthetic cytokine releaser polyriboinosinic-polyribocytidilic acid (poly I:C, 4 mg/kg) on gestational day 15. LI was assessed in 35-day and 3-month-old offspring using a thirst motivated conditioned emotional response procedure. RESULTS: Consistent with the characteristic maturational delay of schizophrenia, prenatal immune activation did not affect LI in the juvenile offspring but led to a post pubertal emergence of LI disruption. In addition, pronounced alterations in hippocampal morphology resembling those found in schizophrenia, were evident in the adult offspring. CONCLUSIONS: These results support the hypothesis that immune activation during pregnancy may lead to long-term abnormalities mimicking those observed in schizophrenia. PMID- 12748760 TI - Comments on "Pediatric intensive care: result of a European survey". PMID- 12748758 TI - Health-related quality of life and symptom experience in tacrolimus-based regimens after renal transplantation: a multicentre study. AB - Tacrolimus is increasingly used as a baseline immunosuppressant after renal transplantation. This multicentre study assessed health-related quality of life and symptom experience in renal transplant patients on tacrolimus-based therapy, using the SF-36 and Euroqol 5 dimensions (EQ-5D) and the 'modified transplant symptom occurrence and symptom distress scale', respectively. Symptoms of depression were assessed with the short form of the Beck Depression Inventory and physical activity with the Baecke questionnaire. Overall, 350 patients with a median post-transplant status of 16.7 months were enrolled. Results revealed that patients experienced lower SF-36 scores than the general population, except in terms of bodily pain. Univariate and multivariate analyses demonstrated that a higher degree of depressive symptoms and female gender were consistently related to a health status perceived as being worse and a higher rate of symptom experience. These findings are in accordance with previous quality-of-life reports that assessed patients under various immunosuppressive therapies. Therefore, interventions, including the screening and treatment for depression and the addressing of gender-specific issues, can enhance quality of life. PMID- 12748762 TI - Effects of native forest regeneration practices on genetic diversity in Eucalyptus consideniana. AB - Impacts of forest harvesting and regeneration practices on genetic diversity in the Australian native forest species Eucalyptus consideniana Maiden (yertchuk) were examined using 29 Mendelian DNA markers (18 RFLPs and 11 microsatellites). Two replicate logging coupes were studied from each of the two most commonly employed silvicultural treatments: clear felling with aerial re-sowing and the seed tree system. For each coupe, genetic diversity measures were compared between a sample of the sapling regeneration and a corresponding control sample from bordering unharvested trees. When calculations were performed over all 29 loci, significant reductions of allelic richness (AR), effective number of alleles (AE) and/or expected heterozygosity (HE) were detected on one or both of seed tree coupes, but on neither of the clear falls. When calculations were performed over the 11 microsatellites alone, all three measures, AR, AE and HE, were significantly reduced on both of the seed replicates but on neither of the two clear falls. In contrast, when the RFLPs were examined separately, there were no significant reductions of diversity on either of the two seed tree coupes or on the two clear falls. These results suggest that genetic erosion is more likely under the seed tree system than under clear-felling with aerial re-sowing and that there is greater statistical power to detect it with microsatellites than with RFLPs. A Monte Carlo simulation to test the statistical significance of the number of apparently lost or gained alleles showed that significant losses of alleles above specified threshold frequencies occurred only in the two seed tree replicates. Three of the four control and regeneration population pairs were significantly differentiated, as indicated by exact tests or by pairwise FST estimates. Comparisons of CONTML dendrograms, constructed for the regeneration populations only versus the control populations only, indicated that genetic drift was significantly promoted under forest management. No significant decreases in observed heterozygosity, or increases in the panmictic index (f), were observed in any of the comparisons suggesting that inbreeding was not promoted by a single rotation of forest management. PMID- 12748763 TI - Construction of a deep coverage BAC library from Capsicum annuum, 'CM334'. AB - A bacterial artificial chromosome (BAC) library consisting of 235,000 clones with an average insert size of 130 kb was constructed from Capsicum annuum, 'CM334'. Based on a pepper haploid genome size of 2,702 Mbp/C, the BAC library is estimated to contain approximately 12 genome equivalents and represents at least 99% of the pepper genome. Screening of the library with mitochondrial DNA probes (coxII, coxIII, atp6 and atp9) and chloroplast DNA probes (atpB, rbcL) indicated that contamination with cytoplasmic DNA was less than 0.5%. To estimate the possibility of isolating a specific clone, the library was screened with single or low-copy gene-specific probes and RFLP probes. Screening of high density BAC filters with RFLP markers linked to L (TMV resistance), y (fruit color), C2 (fruit color) and C (pungency) loci under high stringency conditions revealed that at least three positive BAC clones were found per each probe. This fact indicates that the library is highly reliable and represents a resource for map based cloning, physical mapping, and characterization of upstream and downstream regulations of the chili pepper genes. PMID- 12748764 TI - Brassica napus lines with rearranged Arabidopsis mitochondria display CMS and a range of developmental aberrations. AB - Numerous Brassica napus (+) Arabidopsis thaliana somatic hybrids were screened for male sterility and aberrant flower phenotypes. Nine hybrids were selected and backcrossed recurrently to B. napus. The resulting lines displayed stable maternal inheritance of flower phenotypes. Nuclear and organellar genomes were characterized molecularly using RFLP analysis. No DNA from A. thaliana was found in the nuclear genome after six back-crosses, whilst the mitochondrial genomes contained rearranged DNA from both A. thaliana and B. napus. Each line tested had a unique RFLP pattern of the mitochondrial DNA (mtDNA) that remained unchanged between the BC(3) and BC(6) generation. The plastid genomes consisted of B. napus DNA. Five lines of the BC(5) generation were subjected to more comprehensive investigations of growth, morphology and fertility. On the basis of these investigations, the five CMS lines could be assigned to two groups, one represented by three lines displaying reduced vegetative development, complete male sterility, and homeotic conversions of stamens into feminized structures. The second group, represented by the other two lines, were not completely male sterile but still displayed severely affected flower morphologies. These two lines did not display any reduction in vegetative development. For both groups only stamens and petals suffered from the morphological and functional aberrations, while the sepals and pistils displayed normal morphology. All plants were fully female-fertile. Different rearrangements of the mitochondrial genome disturbed nuclear-mitochondrial interactions and led to various types of aberrant growth and flower development. The existence of numerous CMS lines with different mitochondrial patterns involving a species with a sequenced genome offers new opportunities to investigate the genetic regulation of CMS and its associated developmental perturbations. PMID- 12748765 TI - Unilateral compatibility and genotypic difference in crossability in interspecific hybridization between Dianthus caryophyllus L. and Dianthus japonicus Thunb. AB - Reciprocal interspecific crosses were carried out between six lines of Dianthus caryophyllus L. and one line of Dianthus japonicus Thunb. Although no seed was set when D. japonicus was used as the seed parent, six seedlings were successfully obtained from 2,380 immature ovules by applying the embryo-rescue technique. However, they showed seed parent-like morphology and no evidence for the hybridity by flow cytometry and RAPD analyses. When six lines of D. caryophyllus were used as seed parents, a total of 192 seedlings were successfully obtained without using the embryo-rescue technique. Among these seedlings, 12 out of 25 progenies obtained from the carnation line '98sp1651' were confirmed to be the hybrids. The remaining 13 progenies of this line, and the total 167 progenies obtained from the other carnation lines, had carnation like morphology without any evidence of hybridity by flow cytometry and RAPD analyses. The progenies confirmed as hybrids had intermediate characters of the parents with respect to leaf width and flower size, but they had a uniform flower color, reddish purple, which was different from that of either parent. Since the hybrids obtained in the present study have some profitable characters such as vigorous growth in summer time, upright robust stem, broad leaves and early flowering, they are expected to be used for the breeding of carnation which is suitable for growing under the Japanese climate. PMID- 12748766 TI - Identification of genetically linked RGAs by BAC screening in maize and implications for gene cloning, mapping and MAS. AB - The resistance gene analogue (RGA) pic19 in maize, a candidate for sugarcane mosaic virus (SCMV) resistance gene (R gene) Scmv1, was used to screen a maize BAC library to identify homologous sequences in the maize genome and to investigate their genomic organisation. Fifteen positive BAC clones were identified and could be classified into five physically independent contigs consisting of overlapping clones. Genetic mapping clustered three contigs into the same genomic region as Scmv1 on chromosome 6S. The two remaining contigs mapped to the same region as a QTL for SCMV resistance on chromosome 1. Thus, RGAs mapping to a target region can be successfully used to identify further linked candidate sequences. The pic19 homologous sequences of these clones revealed a sequence similarity of 94-98% on the nucleotide level. The high sequence similarity reveals potential problems for the use of RGAs as molecular markers. Their application in marker-assisted selection (MAS) and the construction of high-density genetic maps is complicated by the existence of closely linked homologues resulting in 'ghost' marker loci analogous to 'ghost' QTLs. Therefore, implementation of genomic library screening, including genetic mapping of potential homologues, seems necessary for the safe application of RGA markers in MAS and gene isolation. PMID- 12748767 TI - Identification of SCAR markers linked to Pl-w mildew resistance in apple. AB - Resistance to powdery mildew is an important objective for cultivar improvement programmes of apple and several different major genes for resistance to mildew are available. Molecular markers linked to such key traits can be used to screen progenies for resistant individuals. A progeny derived from the crab apple 'White Angel' (the source of Pl-w) was screened for resistance to mildew for two seasons in the glasshouse and four seasons in the field. DNA bulks of resistant and susceptible seedlings were screened with 176 AFLP primer combinations. Seven AFLP markers were identified that differentiated the bulks, and two of these markers were developed into SCARs, EM M01 and EM M02, mapping at 4.6 and 6.4 recombination units from Pl-w. PMID- 12748768 TI - Development of PCR-based markers linked to dominant genes for male-fertility restoration in Pampa CMS of rye (Secale cereale L.). AB - Cytoplasmatic male sterility (CMS) is the basis for commercial hybrid seed production of rye. Nuclear restorer genes are indispensable for a complete restoration of fertility of the CMS lines. The drawbacks of current European restorer lines require the utilisation of new genetic resources that have been recently detected in an Iranian primitive rye population (IRAN IX) and an Argentinean landrace (Pico Gentario). The introgression of these effective restorer genes (Rfp1 and Rfp2, respectively) into breeding material can be facilitated by marker-assisted selection. Using two F(2) populations based on crosses between the non-restorer inbred line Lo6 and the restorer IRAN IX, as well as Pico Gentario, RAPDs and AFLPs were screened and led to a closely linked marker set for each of these genes. The conversion of the closest markers into fragment-specific sequence-characterised amplified region (SCAR) markers resulted in flanking ranges of 2.9 cM (Rfp1) and 5.2 cM (Rfp2). The application of these markers in backcross programmes is discussed. PMID- 12748769 TI - Microsatellite genotyping of carnation varieties. AB - A set of 11 sequence-tagged microsatellite markers for carnation (Dianthus caryophyllus) was developed using a DNA library enriched for microsatellites. Supplemented with three markers derived from sequence database entries, these were used to genotype carnation varieties using a semi-automated fluorescence based approach. In a set of 82 cultivars, the markers amplified 4-16 alleles each. The effective number of alleles varied from 1.9 to 6.0. For the eight best scorable markers, heterozygosity was between 0.51 and 0.99. The markers were able to distinguish all cultivars with a unique combination of alleles, except for sport mutants, which were readily grouped together with the original cultivar. In addition, one group of three and one group of six cultivars each had the same combination of 'allelic peaks'. The cluster of three varieties concerned original cultivars and their mutants. The cluster of six consisted of four mutants from the same cultivar and two other varieties. PMID- 12748770 TI - Genetic dissection of pathotype-specific resistance to ascochyta blight disease in chickpea (Cicer arietinum L.) using microsatellite markers. AB - Ascochyta blight is an economically important disease of chickpea caused by the fungus Ascochyta rabiei. The fungus shows considerable variation for pathogenicity in nature. However, studies on the genetics of pathotype-specific resistance are not available for this plant-pathosystem. The chickpea landrace ILC 3279 has resistance to pathotype I and II of the pathogen. In order to understand the inheritance of pathotype-specific resistance in this crop, both Mendelian and quantitative trait loci analyses were performed using a set of intraspecific, recombinant inbred lines derived from a cross between the susceptible accession ILC 1272 and the resistant ILC 3279, and microsatellite markers. We identified and mapped a major locus (ar1, mapped on linkage group 2), which confers resistance to pathotype I, and two independent recessive major loci (ar2a, mapped on linkage group 2 and ar2b, mapped on linkage group 4), with complementary gene action conferring resistance to pathotype II. Out of two pathotype II-specific resistance loci, one (ar2a) linked very closely with the pathotype I-specific resistance locus, indicating a clustering of resistance genes in that region of the chickpea genome. PMID- 12748771 TI - Genetic diversity impacts of forest fires, forest harvesting, and alternative reforestation practices in black spruce (Picea mariana). AB - Benchmarks were established for genetic diversity inherent in natural mature populations, and genetic diversity impacts of forest fires, clearcut harvesting and alternative natural and artificial silvicultural regeneration practices were determined in black spruce (Picea mariana). Allozymes of 32 loci were used to determine and compare genetic diversity and genetic relationships of adjacent or nearby four stand types: post-fire natural mature (FNM), post-fire natural young (FNR), post-harvest natural young (HNR) and post-harvest plantation (PLT), of black spruce at each of the four study sites located in two ecoregions in Manitoba: Ecoregion 90-Lac Seul Upland (Eastern) and Ecoregion 158 - Mid-Boreal Lowland (Northern). Both allelic- and genotypic-based genetic diversity parameters, as well as latent genetic potential, were determined. Black spruce populations showed typical moderate to high levels of allozyme genetic diversity. The mean genetic diversity parameters over the 16 black spruce populations sampled were as follows: percent loci polymorphic - 67%, mean number of alleles per locus - 2.52, effective number of alleles per locus - 1.70, observed heterozygosity - 0.222, expected heterozygosity - 0.308, mean number of observed genotypes per locus - 3.65, mean number of expected genotypes per locus - 5.03, genotype additivity (observed) - 116.8, genotype additivity (expected) - 161, genotype multiplicity (observed) - 6.16 x 10(15), genotype multiplicity (expected) - 2.06 x 10(19) and latent genetic potential - 26.12. The four stand types (FNM, FNR, HNR and PLT) had comparable and statistically similar genetic diversity levels at each of the four study sites as well as overall. No significant differences in black spruce genetic diversity levels were observed between the two ecoregions in Manitoba, as well as between the post-fire and post harvest regenerated stands. No particular order of genetic relatedness among the four stand types was observed. Black spruce populations showed some sort of site related differentiation in their genetic constitution. Allelic heterogeneity and genetic distances among populations within stand types and among four stand types suggest that the genetic diversity was maintained at the landscape level in black spruce. The results of our study demonstrate that forest fires and currently used clearcut harvesting, and alternative natural and artificial silvicultural regeneration practices, do not adversely affect genetic diversity in black spruce, and that the genetic diversity effects of clearcut harvesting are not significantly different from those due to forest fires in black spruce. PMID- 12748772 TI - Molecular linkage maps of Vitis vinifera L. and Vitis riparia Mchx. AB - Two linkage maps for grape (Vitis spp.) have been developed based on 81 F(1) plants derived from an interspecific cross between the wine cultivar Moscato bianco (Vitis vinifera L.) and a Vitis riparia Mchx. accession, a donor of pathogen resistance traits. The double pseudotest-cross mapping strategy was applied using three types of molecular markers. The efficiency of SSRs to anchor homologous linkage groups from different Vitis maps and the usefulness of AFLPs in saturating molecular linkage maps were evaluated. Moreover, the SSCP technique was developed based on sequence information in public databases concerning genes involved in flavonoid and stilbene biosynthesis. For the maternal genetic map a total of 338 markers were assembled in 20 linkage groups covering 1,639 cM, whereas 429 loci defined the 19 linkage groups of the paternal map which covers 1,518 cM. The identification of 14 linkage groups common to both maps was possible based on 21 SSR and 19 AFLP loci. The position of SSR loci in the maps presented here was consistent with other published mapping experiments in Vitis. PMID- 12748773 TI - Frequency of alleles conferring resistance to Bt maize in French and US corn belt populations of the European corn borer, Ostrinia nubilalis. AB - Farmers, industry, governments and environmental groups agree that it would be useful to manage transgenic crops producing insecticidal proteins to delay the evolution of resistance in target pests. The main strategy proposed for delaying resistance to Bacillus thuringiensis ( Bt) toxins in transgenic crops is the high dose/refuge strategy. This strategy is based on the unverified assumption that resistance alleles are initially rare (<10(-3)). We used an F(2) screen on >1,200 isofemale lines of Ostrinia nubilalis Hubner (Lepidoptera: Crambidae) collected in France and the US corn belt during 1999-2001. In none of the isofemale lines did we detect alleles conferring resistance to Bt maize producing the Cry1Ab toxin. A Bayesian analysis of the data indicates that the frequency of resistance alleles in France was <9.20 x 10(-4) with 95% probability, and a detection probability of >80%. In the northern US corn belt, the frequency of resistance to Bt maize was <4.23 x 10(-4) with 95% probability, and a detection probability of >90%. Only 95 lines have been screened from the southern US corn belt, so these data are still inconclusive. These results suggest that resistance is probably rare enough in France and the northern US corn belt for the high-dose plus refuge strategy to delay resistance to Bt maize. PMID- 12748774 TI - The genetics of phytate and phosphate accumulation in seeds and leaves of Arabidopsis thaliana, using natural variation. AB - Phytate (myo-inositol-1,2,3,4,5,6-hexakisphosphate, InsP6) is the most abundant P containing compound in plants, and an important anti-nutritional factor, due to its ability to complex essential micro-nutrients, e.g. iron and zinc. Analysis of natural variation for InsP6 and Pi accumulation in seeds and leaves for a large number of accessions of Arabidopsis thaliana, using a novel method for InsP6 detection, revealed a wide range of variation in InsP6 and Pi levels, varying from 7.0 mg to 23.1 mg of InsP6 per gram of seed. Quantitative trait locus (QTL) analysis of InsP6 and Pi levels in seeds and leaves, using an existing recombinant inbred line population, was performed in order to identify a gene(s) that is (are) involved in the regulation of InsP6 accumulation. Five genomic regions affecting the quantity of the InsP6 and Pi in seeds and leaves were identified. One of them, located on top of chromosome 3, affects all four traits. This QTL appears as the major locus responsible for the observed variation in InsP6 and Pi contents in the L er/Cvi RIL population; the L er allele decreases the content of both InsP6 and Pi in seeds and in leaves. The InsP6/Pi locus was further fine-mapped to a 99-kb region, containing 13 open reading frames. The maternal inheritance of the QTL and the positive correlation between InsP6 and total Pi levels both in seeds and in leaves indicate that the difference in InsP6 level between L er and Cvi is likely to be caused by a difference in transport rather than by an alteration in the biosynthesis. Therefore, we consider the vacuolar membrane ATPase subunit G, located in the region of interest, as the most likely candidate gene for InsP6/Pi. PMID- 12748775 TI - Synthesis of intergeneric hybrids and establishment of genomic affinity between Diplotaxis catholica and crop Brassica species. AB - Intergeneric hybrids of the wild crucifer Diplotaxis catholica (2n = 18, D(C)D(C)) as female with two crop Brassica species, namely Brassica rapa (2n = 20; AA) and Brassica juncea (2n = 36; AABB) as male, were developed, using ovary and sequential culture. Reciprocal crosses were not successful, suggesting unilateral cross incompatibility. Morphologically, the hybrid plants resembled the crop brassica parents, but were nearly male- as well as female-sterile. Induction of amphiploidy helped to improve pollen fertility for the D. catholica x B. rapa cross (73%), but less so for the D. catholica x B. juncea cross (35 40%). Female fertility was also higher in both the amphiploids. Cytological analysis of the F(1) hybrids revealed aberrant meiosis with predominant occurrence of the univalents. Partial genomic homoeology between the A genome of B. rapa and the D(C) genome of D. catholica was indicated by the presence of up to five bivalents in 14.7% of the PMCs in the D. catholica x B. rapa hybrid, and 1-2 trivalents or a quadrivalent in nearly 44% of the PMCs in the derived amphiploid. In the second cross, D. catholica x B. juncea, up to six bivalents and one trivalent were observed indicating homoeology between the A/B genomes of B. juncea and the D(C) genome of D. catholica. The possibility of introgression of desirable genes from D. catholica into crop Brassica species exists in view of significant affinity between the D(C) and A/B genomes. PMID- 12748776 TI - A new Hessian fly resistance gene (H30) transferred from the wild grass Aegilops triuncialis to hexaploid wheat. AB - A new Hessian fly (Mayetiola destructor Say) resistance gene from Aegilops triuncialis and its transfer to hexaploid wheat via interspecific hybridisation is described. The transfer line TR-3531 (42 chromosomes), derived from the cross [(Triticum turgidum x Ae. triuncialis) x Triticum aestivum] and carrying the Heterodera avenae resistance gene Cre7, showed a high level of resistance to the M. destructor biotype prevailing in the SW of Spain. A single dominant gene (H30) seems to determine the Hessian fly resistance in this introgression line, and its linkage with an isozyme marker (Acph-U1) has also been studied. It has been demonstrated that the resistance gene H30 in the TR-3531 line is non-allelic with respect to the genes H3, H6, H9, H11, H12, H13, H18 and H21, present in wheat cultivars from the Uniform Hessian Fly Nursery (UHFN), as well as to H27, carried by the introgression line H-93-33. Advanced lines with the H30 gene were obtained by backcrossing the transfer line and different commercial wheats as recurrent parents. Several of them showed a high yield in tests carried out in the infested field. Electronic Supplementary Material is available if you access this article at http://dx.doi.org/10.1007/s00122-002-1182-z. On that page (frame on the left side), a link takes you directly to the supplementary material. PMID- 12748777 TI - Estimates of conserved microsynteny among the genomes of Glycine max, Medicago truncatula and Arabidopsis thaliana. AB - A growing body of research indicates that microsynteny is common among dicot genomes. However, most studies focus on just one or a few genomic regions, so the extent of microsynteny across entire genomes remains poorly characterized. To estimate the level of microsynteny between Medicago truncatula (Mt) and Glycine max (soybean), and also among homoeologous segments of soybean, we used a hybridization strategy involving bacterial artificial chromosome (BAC) contigs. A Mt BAC library consisting of 30,720 clones was screened with a total of 187 soybean BAC subclones and restriction fragment length polymorphism (RFLP) probes. These probes came from 50 soybean contig groups, defined as one or more related BAC contigs anchored by the same low-copy probe. In addition, 92 whole soybean BAC clones were hybridized to filters of HindIII-digested Mt BAC DNA to identify additional cases of cross-hybridization after removal of those soybean BACs found to be repetitive in Mt. Microsynteny was inferred when at least two low-copy probes from a single soybean contig hybridized to the same Mt BAC or when a soybean BAC clone hybridized to three or more low-copy fragments from a single Mt BAC. Of the 50 soybean contig groups examined, 54% showed microsynteny to Mt. The degree of conservation among 37 groups of soybean contigs was also investigated. The results indicated substantial conservation among soybean contigs in the same group, with 86.5% of the groups showing at least some level of microsynteny. One contig group was examined in detail by a combination of physical mapping and comparative sequencing of homoeologous segments. A TBLASTX similarity search was performed between 1,085 soybean sequences on the 50 BAC contig groups and the entire Arabidopsis genome. Based on a criterion of sequence homologues <100 kb apart, each with an expected value of < or =1e-07, seven of the 50 soybean contig groups (14%) exhibited microsynteny with Arabidopsis. PMID- 12748778 TI - Rapid identification of 1-aminocyclopropane-1-carboxylate (ACC) synthase genotypes in cultivars of Japanese pear (Pyrus pyrifolia Nakai) using CAPS markers. AB - In Japanese pear (Pyrus pyrifolia Nakai), fruit storage potential is closely related to the amount of ethylene produced. We have developed a rapid and accurate method for analyzing genes involved in high ethylene production during fruit ripening in Japanese pear. This involves cleaved-amplified polymorphic sequences (CAPS) of two 1-aminocyclopropane-1-carboxylate (ACC) synthase genes (PPACS1 and PPACS2). Two CAPS markers (A for PPACS1 and B for PPACS2), associated with the amount of ethylene produced, were identified. Marker A was associated with high ethylene producers and marker B with moderate ethylene producers. The absence of these two markers enabled the identification of low ethylene producers. Using these markers, we have identified ethylene genotypes for 40 Japanese pear cultivars and two Chinese pear (P. bretschneideri) cultivars that are commercially important and used in breeding programs. Furthermore, we performed linkage analysis of these two genes in the F(2) population, which revealed that the recombination frequency between the two markers was 20.8 +/- 3.6%. This information is critical to the selection of parents and in breeding strategies to improve storage ability of Japanese pears. PMID- 12748779 TI - A first linkage map of olive (Olea europaea L.) cultivars using RAPD, AFLP, RFLP and SSR markers. AB - The first linkage map of the olive (Olea europaea L.) genome has been constructed using random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphisms (AFLP) as dominant markers and a few restriction fragment length polymorphisms (RFLP) and simple-sequence repeats (SSR) as codominant markers. Ninety-five individuals of a cross progeny derived from two highly heterozygous olive cultivars, Leccino and Dolce Agogia, were used by applying the pseudo test cross strategy. From 61 RAPD primers 279 markers were obtained - 158 were scored for Leccino and 121 for Dolce Agogia. Twenty-one AFLP primer combinations gave 304 useful markers - 160 heterozygous in Leccino and 144 heterozygous in Dolce Agogia. In the Leccino map 249 markers (110 RAPD, 127 AFLP, 8 RFLP and 3 SSR) were linked. This resulted in 22 major linkage groups and 17 minor groups with fewer than four markers. In the Dolce Agogia map, 236 markers (93 RAPD, 133 AFLP, 6 RFLP and 4 SSR) were linked; 27 major linkage groups and three minor groups were obtained. Codominant RFLPs and SSRs, as well as few RAPDs in heteroduplex configuration, were used to establish homologies between linkage groups of both parents. The total distance covered was 2,765 cM and 2,445 cM in the Leccino and Dolce Agogia maps, respectively. The mean map distance between adjacent markers was 13.2 cM in Leccino and 11.9 cM in Dolce Agogia, respectively. Both AFLP and RAPD markers were homogeneously distributed in all of the linkage groups reported. The stearoyl-ACP desaturase gene was mapped on linkage group 4 of cv. Leccino. PMID- 12748780 TI - QTL mapping provides evidence for lack of association of the avoidance of leaf rust in Hordeum chilense with stomata density. AB - In cereals, rust fungi are among the most harmful pathogens. Breeders usually rely on short-lived hypersensitivity resistance. As an alternative, "avoidance" may be a more durable defence mechanism to protect plants to rust fungi. In Hordeum chilense avoidance is based on extensive wax covering of stomata, which interferes with the induction of appressorium formation by the rust fungi. High avoidance levels are associated with a higher stoma density on the abaxial leaf epidermis. The avoidance level was assessed as the percentage of germ tube/stoma encounters that did not result in appressorium differentiation by Puccinia hordei, the barley leaf rust fungus. One hundred F(2) individuals from the cross between two H. chilense accessions with contrasting levels of avoidance showed a continuous distribution for avoidance of the rust fungus and for stoma density, indicating quantitative inheritance of the traits. No significant correlation was found between avoidance and stoma density in the segregating F(2) population. In order to map quantitative trait loci (QTLs) for both traits, an improved molecular marker linkage map was constructed, based on the F(2) population. The resulting linkage map spanned 620 cM and featured a total of 437 AFLP markers, thirteen RFLPs, four SCARs, nine SSRs, one STS and two seed storage protein markers. It consisted of seven long and two shorter linkage groups, and was estimated to cover 81% of the H. chilense genome. Restricted multiple interval mapping identified two QTLs for avoidance and three QTLs for stoma density in the abaxial leaf surface. The QTLs for avoidance were mapped on chromosome 3 and 5; those for stoma density on chromosomes 1, 3 and 7. Only the two QTLs regions located on chromosome 3 (one for avoidance and the other for stoma density) overlapped. The wild barley H. chilense has a high crossability with other members of the Triticeae tribe. The knowledge on the location of the QTLs responsible for the avoidance trait is a prerequisite to transfer this favourable agronomic trait from H. chilense to cultivated cereal genomes. PMID- 12748782 TI - Molecular mapping of genomic regions underlying barley yellow dwarf tolerance in cultivated oat (Avena sativa L.). AB - Barley yellow dwarf (BYD) is one of the most important viral diseases in small grains, including oat (Avena sativa L.). Breeding for BYD tolerance is an effective and efficient means to control the disease. Characterization of major sources of tolerance, and identification of marker and the trait associations, will directly benefit breeding for BYD tolerance. Genomic regions underlying BYD tolerance were mapped and characterized in an oat population consisting of 152 recombinant inbred lines from the cross of 'Ogle' (tolerant)/MAM17-5 (sensitive). Tolerance was evaluated in replicated field trials across 2 years under artificial inoculation with viruliferous aphids harboring BYD virus isolate PAV IL. Composite interval mapping was used for quantitative trait loci (QTLs) analysis with a framework map consisting of 272 molecular markers. Four QTLs, BYDq1, BYDq2, BYDq3 and BYDq4, for BYD tolerance were identified on linkage groups OM1, 5, 7 and 24, respectively. All but BYDq2 were consistently detected across both years. Significant epistasis was found between some QTLs. The final model including the epistatic effect explained 50.3 to 58.2% of the total phenotypic variation for BYD tolerance. Some QTLs for BYD tolerance were closely linked to QTLs for plant height and days to heading. Potential problems with QTL mapping for BYD tolerance have been discussed. The identified association of markers and tolerance should be useful to pyramid favorable alleles for BYD tolerance into individual oat lines. PMID- 12748781 TI - Identification of AFLP and STS markers closely linked to the def locus in pea. AB - The recessive mutation of the def gene of pea (Pisum sativum L.) leads to the loss of the hilum, the abscission zone between the seed and the pod. Thereby, it reduces the free dispersal of the seeds through pod shattering. As a prerequisite for a gene isolation via a map-based cloning approach, bulked segregant analysis followed by single plant analyses of over 200 homozygous individuals of a population of 476 F2 plants derived from a cross between 'DGV' (def wild-type) and 'PF' (def mutant), were used to detect markers closely linked to the def locus. The AFLP technique in combination with silver staining was used to maximize numbers of reproducible marker loci. Fifteen AFLP loci showed a genetic distance less than 5 and two of them less than 1 centiMorgans (cM) to the gene of interest. AFLPs were converted into sequence tagged sites (STSs) and into a newly refined AFLP-based single locus marker named the 'sequence specified AFLP' (ssAFLP). PMID- 12748783 TI - Intra-clonal variation and a similarity threshold for identification of clones: application to Salix exigua using AFLP molecular markers. AB - Although molecular methods are a major advance over phenological or root connectivity studies in the identification of clonal plants, there is still a level of ambiguity associated with two types of error: misidentification of genetically similar seedlings as clones and misidentification of dissimilar fingerprints from clones as genetically distinct individuals. We have addressed the second of these error types by determining the level of variation for AFLP fingerprints in Salix exigua, and then by developing a threshold value of Jaccard's similarity index for assigning individuals to clones or to siblings. Variation in AFLP banding patterns among clones was partitioned into three potential sources; clones, stems within-clones and foliage within-stems. Most of the variation was attributable to clones and then to stems within-clones. To provide an objective means of identifying clones, we developed a method for establishing a threshold similarity index to assign individuals to the same clone. Our method yielded a Jaccard similarity threshold of 0.983 that resulted in a potential pairwise error rate of 8.1% putative clone assigned to siblings and 1.5% sibling assigned to clones. The method was tested on independent clonal and sibling individuals resulting in the same threshold value and similar error rates. We applied our method to assign individuals to clones in a population of S. exigua along the Cosumnes River, California. A total of 11 clones were identified, with one clone including 43% of the individuals sampled. Our results show that this approach can be useful in the accurate identification of clones. PMID- 12748784 TI - Comparative analysis on the genetic relatedness of Sorghum bicolor accessions from Southern Africa by RAPDs, AFLPs and SSRs. AB - In order to get an overview on the genetic relatedness of sorghum (Sorghum bicolor) landraces and cultivars grown in low-input conditions of small-scale farming systems, 46 sorghum accessions derived from Southern Africa were evaluated on the basis of amplified fragment length polymorphism (AFLPs), random amplified polymorphic DNAs (RAPDs) and simple sequence repeats (SSRs). By this approach all sorghum accessions were uniquely fingerprinted by all marker systems. Mean genetic similarity was estimated at 0.88 based on RAPDs, 0.85 using AFLPs and 0.31 based on SSRs. In addition to this, genetic distance based on SSR data was estimated at 57 according to a stepwise mutation model (Deltamu-SSR). All UPGMA-clusters showed a good fit to the similarity estimates (AFLPs: r = 0.92; RAPDs: r = 0.88; SSRs: r = 0.87; Deltamu-SSRs: r = 0.85). By UPGMA clustering two main clusters were built on all marker systems comprising landraces on the one hand and newly developed varieties on the other hand. Further sub-groupings were not unequivocal. Genetic diversity (H, DI) was estimated on a similar level within landraces and breeding varieties. Comparing the three approaches to each other, RAPD and AFLP similarity indices were highly correlated (r = 0.81), while the Spearman's rank correlation coefficient between SSRs and AFLPs was r = 0.57 and r = 0.51 between RAPDs and SSRs. Applying a stepwise mutation model on the SSR data resulted in an intermediate correlation coefficient between Deltamu-SSRs and AFLPs (r = 0.66) and RAPDs ( r = 0.67), respectively, while SSRs and Deltamu-SSRs showed a lower correlation coefficient (r = 0.52). The highest bootstrap probabilities were found using AFLPs (56% on average) while SSR, Deltamu-SSR and RAPD-based similarity estimates had low mean bootstrap probabilities (24%, 27%, 30%, respectively). The coefficient of variation (CV) of the estimated genetic similarity decreased with an increasing number of bands and was lowest using AFLPs. PMID- 12748786 TI - [Mesenteric ischemia. Surgical epidemiology--when to take it into consideration?]. AB - Diagnostic and therapeutic treatment modalities have developed over the last few years. However, the mortality rate of mesenteric ischemia is still high. The incidence amounts to 2-4 patients in 100,000 inhabitants. This is due to the fact that disturbances in the blood circulation of the intestine are not recognized early enough. While clinical symptoms are different during the acute and chronic course of mesenteric ischemia, the diagnostic and therapeutic consequences are similar. In order to decrease the mortality rate, it is important to keep mesenteric ischemia in mind when confronted with abdominal pain in elderly patients. This is particularly important as life expectancy in the western population is increasing. PMID- 12748785 TI - AFLP genetic polymorphism in wild barley (Hordeum spontaneum) populations in Israel. AB - The genetic diversity produced by the amplified fragment length polymorphism (AFLP) method was studied in 94 genotypes of wild barley, Hordeum spontaneum (C. Koch) Thell., originating from ten ecologically and geographically different locations in Israel. Eight primer pairs produced 204 discernible loci of which 189 (93%) were polymorphic. Each genotype had a unique banding profile and the genetic similarity coefficient varied between 0.74 and 0.98. The phenogram generated from these similarities by the UPGMA method did not group genotypes strictly according to their geographical origin, which pattern was also seen in the principal coordinate (PCO) plot. Genetic diversity was larger within (69%) than among (31%) populations. Associations between ecogeographical variables and the mean gene diversity were found at one primer pair. The results are discussed and compared with data obtained by the simple sequence repeat (SSR) method. PMID- 12748787 TI - [Emergency diagnostic imaging in mesenteric ischemia]. AB - Diagnostic imaging in patients with suspected acute mesenteric ischemia is started with abdominal ultrasound including duplex sonography of the mesenteric vessels. Despite low sensitivity even in experienced hands, ultrasound is used because operative treatment can be initiated without further imaging if a positive diagnosis is made. Plain abdominal X-rays are usually unspecific in acute mesenteric ischemia and are mainly used to rule out differential diagnoses. Spiral CT (ideally using a multislice technique) can accurately demonstrate morphology of the arterial and venous mesenteric vessels, changes in the bowel wall, and additional mesenteric or peritoneal findings. Therefore, CT has the potential for diagnosis of mesenteric ischemia on a pathological basis. Furthermore, CT is successfully used to confirm or exclude most other causes of acute abdominal conditions. Magnetic resonance imaging (MRI) may be as accurate as CT for the diagnosis of acute mesenteric ischemia and its differential diagnoses. However, MRI is not widely available and therefore not used as an emergency imaging modality so far. Catheter angiography remains the diagnostic gold standard for mesenteric vasculature when spiral CT is not available. PMID- 12748788 TI - [Mesenteric ischemia. Is diagnostic laparoscopy of value?]. AB - In the past decade laparoscopic surgery replaced many open operations in general surgery. Apart from therapeutic uses in cholecystectomy, appendectomy, hernia surgery, gastric fundoplication, and increasingly also large intestine surgery, it is indicated diagnostically first of all for unclear abdominal findings and for staging of intra-abdominal malignancies. To date laparoscopy has been used occasionally for diagnosis and therapy of mesenteric ischemia. Patients suffering from mesenteric ischemia are usually old and have comorbid conditions. Quick diagnosis and therapy are necessary due to the pathogenesis of the disease. The low rate of morbidity as well as the easy availability of laparoscopy in principle favor the employment of laparoscopy also for mesenteric ischemia. Against the background of increasing experience in the area of laparoscopic surgery, this study gives an overview of the present value of laparoscopy for mesenteric ischemia. PMID- 12748789 TI - [Partial ischemia. Occlusive and nonocclusive mesenteric ischemia, ischemic colitis, systemic lupus erythematosus]. AB - Partial mesenteric ischemia is defined as an incomplete occlusion of the superior mesenteric artery resulting in acute abdominal pain, distended abdomen,and bowel hypomotility on auscultation. This disease can be acute or chronic and is caused by vascular occlusion or non-occlusive mechanisms. CT scan and ultrasound show a thickening of the ischemic bowel wall. On endoscopy, initially mucosal edema is observed which may proceed to necrosis. Therapy modalities depend upon the clinical findings: prevailing acute abdominal pain and peritonitis result in emergency laparotomy; prevailing cramping abdominal pain without clinical signs of peritonitis allows time for further diagnostic steps such as mesenteric angiography and interventional procedures. Laparoscopy should be performed in exceptional situations only. PMID- 12748790 TI - [Acute mesenteric ischemia. Resection or reconstruction?]. AB - Acute mesenteric ischemia represents an intensive medical emergency which, when untreated, leads to sepsis and multiorgan failure. Predictive factors for survival are the time duration between onset of symptoms and therapy, etiology,patient age, and immediate therapy with anticoagulants in case of acute mesenteric thrombosis. Pathologically raised laboratory parameters (lactate, acidosis, leukocytosis) are signs of advanced mesenteric ischemia. Therapy aims at reinstating arterial perfusion of ischemic and potentially life-threatening intestinal regions and at avoiding further ischemias or infarctions. Immediate vessel reconstruction (embolectomy, thrombectomy,TEA,bypass) can prevent mesenteric infarction and is therefore preferable to intestinal resection alone, as long as irreversible ischemia or sepsis are not present. If the degree of reperfusion is in question, the indication for "second-look laparotomy" is to be used generously. PMID- 12748792 TI - [Is primary total thyroidectomy justified in benign multinodular goiter? Results of a prospective quality assurance study of 45 hospitals offering different levels of care]. AB - INTRODUCTION: After subtotal resection of multinodular goiter, rates of up to 40% are reported for recurrent goiter in the long-term follow-up. Because of the increased morbidity of surgery for recurrent goiter, this study evaluated the preconditions that would justify total thyroidectomy as part of the primary therapy concept for benign multinodular goiter. MATERIAL AND METHODS: The Quality Assurance Study of Benign and Malignant Goiter covering the period from 1 January to 31 December 1998 assessed 5195 patients treated for benign goiter by primary bilateral resection. With respect to the extent of resection three groups were analyzed: bilateral subtotal resection (ST+ST, n=4580), subtotal resection with contralateral lobectomy (ST+HT, n=527), and total thyroidectomy (TT, n=88). RESULTS: The age of the patients was significantly higher (60.3 years) in the TT group than in the ST+ST (52.5 years) and ST+HT (55.6 years) groups. ASA classification grades III and IV were significantly more frequent in the TT group. The postoperative morbidity increased with the extent of resection. The rate of permanent recurrent laryngeal nerve (RLN) palsy was 0.8% for ST+ST, 1.4% for ST+HT, and 2.3% for TT and of permanent hypoparathyroidism 1.5% for ST+ST, 2.8% for ST+HT, and 12.5% for TT. Multivariate analysis showed that the extent of resection significantly increased the risk of RLN palsy (transient RR 0.5, permanent RR 0.4) and hypoparathyroidism (transient RR 0.2,permanent RR 0.08). The surgeon's experience (RR 0.6) and identification of the RLN (RR 0.5) reduced the risk of permanent RLN palsy. Additionally, the development of permanent hypoparathyroidism was reduced if at least two parathyroid glands (RR 0.4) were identified. CONCLUSION: Total thyroidectomy is associated with an increased rate of RLN palsies and hypoparathyroidism in comparison to less extensive thyroid surgery. In the hands of well-trained surgeons using an appropriate intraoperative technique, primary thyroidectomy is justified if the patient has an increased risk of recurrent goiter. Due to the increased postoperative morbidity after total thyroidectomy, subtotal thyroid resection based on the morphologic changes in the thyroid gland is still recommended as the standard treatment regimen for multinodular goiter. PMID- 12748791 TI - [Descending intramedullary nailing for the treatment of displaced supracondylar humeral fractures in children]. AB - BACKGROUND: Supracondylar humeral fractures are the most common lesions in childhood. Severely displaced fractures are commonly treated by crossed K-wire osteosynthesis. Such stabilized fractures require a cast and often involve postoperative complications such as iatrogenic lesions of the n. ulnaris and secondary displacements,sometimes leading to a consecutive cubitus varus. This study analyzed possible advantages of elastic stabile intramedullary nailing (ESIN) with postoperative release of motion. PATIENTS AND METHOD: All children aged 1-14 years suffering from a supracondylar humeral fracture, with a displacement in at least two planes,were included in this study (period: 1 June 1999-30 April 2001). Movement was permitted for all patients postoperatively. A follow-up examination was scheduled at least 6 months after trauma. RESULTS: This study included 20 female and 30 male patients. Neither iatrogenic lesions of the n. ulnaris nor secondary displacements occurred. Five slight technical pitfalls (perforating wire, gapping with consecutive axis deviation) occurred. Postoperative X-rays most often revealed a remaining axis deviation in the sagittal plane. Comminuted fractures with corresponding zones resulted in cubiti vari and valgi. Of the 50 patients, 47 appeared for follow-up (94%). The ROM coincided with the acquired radiological data. Only one patient (2%) showed a functional deficit greater than 10 degrees. CONCLUSION: As soft tissue swelling does not hinder the surgeon, ESIN shows a high rate of closed stabilizations. No cast has to be applied and free ROM can be permitted for all patients. Avoidance of iatrogenic lesions of the n. ulnaris and secondary displacements are other advantages of this method. PMID- 12748793 TI - [Indicators of quality in surgical treatment of rectal carcinoma]. AB - INTRODUCTION: For the assessment of surgical therapy for rectal cancer, perioperative and oncological as well as aspects of quality of life have to be taken into consideration. The objective of the present analysis was to identify indicators allowing assessment of the quality of the surgical intervention. PATIENTS AND METHODS: Between March 1997 and August 1998,50 patients with adenocarcinoma of the rectum were operated on by five surgeons according to the concept of total or partial mesorectal excision. In 4 patients an anterior, in 35 a low anterior resection, and in 11 an abdominoperineal resection was performed. There was a stage IV (UICC) present in 9 cases, and in 18 patients the tumor extension was restricted to the wall. The demographic and perioperative data as well as the results of the follow-up examination were registered prospectively. The median follow-up period amounted to 44 months (5-57). RESULTS: Intraoperative parameters showed no differences concerning the individual operating surgeons. General complications arose in 9 (50%) and surgical complications in 34 (68%) patients. Anastomotic leakage (5/39), perineal wound infection, and bladder dysfunction (requiring catheterization for bladder emptying) occurred as frequent complications and there were differences with regard to the surgeon. The rate of local recurrence was different and was between 0% and 75% for the individual surgeons. CONCLUSIONS: For the assessment of surgery for rectal carcinoma, only a few parameters are necessary. As indicators of quality after rectal resection, the rate of anastomotic leakage should be registered; after abdominoperineal resection, the rate of perineal wound infection, the ratio of postoperative bladder dysfunction, and the locoregional rate of recurrence should be registered. PMID- 12748794 TI - [Toxic epidermal necrolysis. A case for the burn intensive care unit]. AB - Toxic epidermal necrolysis is a rare but life-threatening exfoliating disease of the skin and mucous membranes, commonly considered a drug sensitivity reaction. In this review of the literature we discuss the epidemiology, etiology, and pathology. We show diagnostic problems and current treatment strategies. Many of the problems associated with the disease, such as wound infection, sepsis, nutritional support, and pain management, are similar to problems in severely burned patients. Burn centers provide optimal logistics and knowledge to diagnose and treat this serious disease entity. PMID- 12748795 TI - [Primary midline closure after excision of a pilonidal sinus is associated with a high recurrence rate]. AB - BACKGROUND: There is a high incidence of postoperative complications and late recurrences after operative therapy of a pilonidal sinus. The optimal treatment strategy is still matter of discussion. We studied the long-term results after excision of a pilonidal sinus and primary midline closure compared with the open surgical procedure. MATERIALS AND METHODS: A total of 73 patients (62 male and 11 female, mean age 26.6 years) underwent a total of 79 operations between 1992 and 2001. Thirty patients (38%) were previously operated on because of a pilonidal sinus.Twenty-four were treated in our institution by an open procedure (five after simple abscess incision, 19 after sinus excision) and 52 by primary midline closure. Another three patients received skin flap procedures. RESULTS: Follow-up was possible for 65 patients (82%) for a median of 50 months.Recurrent pilonidal sinus occurred in 22 cases: 18 after primary midline closure (42%) and four after open procedure (21%, P=0.4). We found no relapse following the three skin flap procedures. There was a significantly higher relapse rate in patients operated with recurrent disease (12/25 vs 10/40; P<0.05). CONCLUSIONS: Despite of numerous previously operated patients (38%), there was a high recurrence rate (42%) after excision of a pilonidal sinus and primary midline closure. Alternative operative techniques creating a lateral wound or the various skin flap procedures may be promising alternatives. We are in the process of changing our treatment strategy for patients suffering from a pilonidal sinus. PMID- 12748796 TI - [Hyperaldosteronism persisting after subtotal adrenalectomy]. AB - BACKGROUND: Primary aldosteronism is known to be caused by aldosterone-producing adenoma (APA). Total adrenalectomy is the standard procedure. In contrast to bilateral adrenal diseases (e.g., MEN II pheochromocytomas), there is no consensus about the effect of subtotal adrenalectomy. CASE REPORT: A 44-year-old patient with primary aldosteronism caused by APA underwent subtotal adrenalectomy including removal of one adenoma. Because hypertension and hypokalemia did not disappear and hyperaldosteronism persisted, the patient had to undergo reoperation in which the adrenalectomy was completed. DISCUSSION: Subtotal adrenalectomy in patients with Conn's syndrome is an interesting therapeutic option,whereas its effect is much higher in hereditary diseases of the adrenal gland. The benefit of preserved adrenal tissue has to be weighed against a possible persistence of hyperaldosteronism, especially in cases with normal opposite adrenal glands. PMID- 12748797 TI - [Tuberculosis of the gall bladder]. AB - We report a case of gall bladder tuberculosis in a 64-year-old male. The gall bladder is an extremely rare localization of an infectious disease seen frequently worldwide--tuberculosis. The reason for this special resistance against the mycobacteria is not clear and is controversial. In imaging, the disease can mimic acute or chronic cholecystitis or carcinoma of the gall bladder. It is important to consider tuberculous cholecystitis in differential diagnosis and to do tuberculin skin tests in case of suspicion. This test is technically easy and cost-effective. Since 1968, isolated tuberculosis of the gall bladder has not been reported in western civilization. This case study was done because of the rareness of the disease,and we review the literature on this topic. PMID- 12748798 TI - Plasma endothelin-1 levels in patients with biliary atresia: possible role in development of portal hypertension. AB - BACKGROUND: Biliary atresia (BA) is a severe neonatal liver disease characterized by progressive extrahepatic biliary tract and intrahepatic inflammatory process. Hepatic fibrosis and portal hypertension (PH) still occur despite the disappearance of jaundice following successful hepatic portoenterostomy. Endothelin-1 (ET-1) is a potent vasoconstrictor and has been reported to stimulate hepatic collagen synthesis. The aim of this study was to demonstrate the potential role of ET-1 in the pathogenesis of the progressive inflammation, fibrosis and PH in BA. METHODS: Thirty pediatric patients with biliary atresia post-hepatic portoenterostomy and 12 healthy children were examined. The ET-1 level was determined by commercially available enzyme-linked immunosorbent assay kits. RESULTS: Endothelin-1 levels were elevated in the patients compared with those of the controls (5.45+/-3.34 vs. 2.74+/-2.17 pg/ml, P = 0.01). Moreover, patients with PH also had greater levels of ET-1 than those without PH (6.73+/ 3.27 vs. 3.26+/-2.2 pg/ml, P = 0.004). Patients with abnormal transaminase enzymes had significantly higher ET-1 levels than those with normal enzymes (6.43+/-3.33 vs. 3.17+/-2.1 pg/ml, P = 0.01). In the jaundice-free group, endothelin-1 levels were elevated in the patients with PH compared with those without PH (5.93+/-2.15 vs. 2.88+/-2.1 pg/ml, P = 0.02). CONCLUSIONS: Our findings showed elevation of plasma ET-1 levels in patients with BA, especially in those with PH. ET-1 levels were also higher in patients with elevated transminase enzymes as well as in the jaundice-free group with PH. ET-1 might play a role in the pathogenesis of the progressive inflammation, fibrosis and PH in BA. PMID- 12748799 TI - Small bowel perforation in the premature neonate: congenital or acquired? AB - To determine the potential aetiological factors of small bowel perforation in the premature neonate, we performed a retrospective chart review of those neonates with spontaneous intestinal perforation (SIP) of the small bowel seen in our tertiary paediatric hospital between January 1980 and December 2000. Data were collected on gestational feto-maternal health, medical interventions prior to perforation and the subsequent operative and laboratory findings. There were 23 patients with SIP of the small bowel over the 21-year review; 65% were male. There were 7 twin pregnancies but no cases linked to maternal drug abuse. The median gestational age was 27 weeks, the median birth weight 973 g, 19 neonates required ventilation, 15 steroids and 13 indomethacin. The median age at diagnosis was 7 days, heralded by rapid development of abdominal distension in 22 patients. Surgical intervention in addition to insertion of a peritoneal drain was required in 19 patients. Positive microbiological cultures of blood or peritoneal fluid at operation were documented in 8 patients; 5 grew Staphylococcus epidermidis and 4 Candida species. Perforations were located in the ileum in 20 and the jejunum in 1. Deficiency of the muscularis propria was found in 6 patients. Of the 6 deaths, 2 neonates had significant co-morbidity in addition to extreme prematurity. Small bowel SIP occurs in the premature neonate after the first week of life and usually presents with abdominal distension. Putative risk factors identified included twin gestation, neonatal ventilation, use of steroids and indomethacin, infection with Staphylococcus epidermidis and Candida species and deficiency of enteric smooth muscle. PMID- 12748800 TI - Pierre-Robin syndrome associated with Chiari type I malformation. AB - CASE REPORT: Pierre-Robin syndrome (PRS) is a rare congenital malformation that shows severe micrognathia and cleft soft palate. A 15-year-old boy who was admitted with occipital headache and gait disturbance was diagnosed with PRS. Radiological evaluation revealed severe herniation of the cerebellar tonsil and multiple craniovertebral osseous anomalies. We carried out foramen magnum decompression (FMD) with duroplasty. Postoperative MRI showed the cerebellar tonsil was freed from strangulation. RESULT AND CONCLUSION: This patient presented with the common finding between PRS and Chiari type I on the embryological aspect by illustrating pathophysiology of the Chiari I malformation. To our knowledge, this is the first reported case of PRS associated with Chiari malformation type I. PMID- 12748801 TI - [Diabetic retinopathy. Pathophysiology and therapy of hypoxia-induced inflammation]. AB - Diabetic retinopathy is the most common chronic microvascular complication associated with diabetes mellitus. The development of diabetic retinopathy is a consequence of metabolic dysregulation. Hyperglycemia is a critical factor which is involved in basement membrane thickening, loss of pericytes and endothelial cells, and retinal capillary nonperfusion. We review the molecular basis of diabetic retinopathy and maculopathy and elaborate the role of growth factors and cytokines in the development of diabetic vascular alterations, their specific influence on the cellular interaction between retinal endothelial cells and pericytes, and the role of intravascular blood components. PMID- 12748802 TI - [Angioproliferative retinal disease caused by ischemia]. AB - Ischemia is a major stimulus for angiogenesis, a biological response mechanism that describes the formation of new blood vessels from existing vessels. An ischemic cell communicates with endothelial cells by soluble factors such as VEGF (vascular endothelial growth factor) and its receptors. A major transcriptional factor for VEGF is HIF-1 (hypoxia inducible factor). Proliferation of endothelial cells alone does not result in stable vascular tubes, this is only achieved by recruiting additional cells such as pericytes. The stabilisation and destabilisation of vessels, which are important prerequisites for vascular growth, are in a dynamic equilibrium which can be modified by additional growth factors such as angiopoietins. In this review we discuss some of the molecular mechanisms leading from ischemia to proliferative retinopathy with a special focus on retinopathy of prematurity and the closely related mouse model of hyperoxia-induced retinopathy. This model is very useful when developing new antiangiogenic therapies based on the increasing understanding of the molecular pathogenesis of ischemic proliferative retinopathy. PMID- 12748803 TI - [Angiographic-histological correlation of late exudative age-related macular degeneration]. AB - AIMS: To correlate the fluorescein angiographic (FAG) classification in age related macular degeneration (AMD) with the histology of surgically excised choroidal neovascularization (CNV) membrane specimens. METHODS: 36 subretinal lesions surgically excised during macular translocation were investigated by light microscopy. Histologic results were correlated with fluorescein angiographic findings. RESULTS: Classic CNV lesions showed a subretinal fibrovascular membrane on histological examination. About 50% of the investigated classic CNV lesions had additional subpigment epithelial components, too. Occult CNV lesions showed a subpigment epithelial fibrovascular membrane on histological examination. A common histological component in lesions of exudative age-related macula degeneration was the appearance of diffuse deposits at the choroidal side of the retinal pigment epithelium. CONCLUSION: Different FAG characteristics in exudative AMD lesions were comparable with the histological results. In correspondence with FAG classification the CNV membranes were localized in different positions in relation to the RPE. PMID- 12748804 TI - [Photodynamic therapy: extended indication]. AB - OBJECTIVE: Evaluation of efficacy of photodynamic therapy (PDT) for classic choroidal neovascularization (CNV) beyond the common indication. METHODS: A group of 256 patients was treated with photodynamic therapy from January to December 2000. Ten of these patients had a predominantly classic CNV due to angioid streaks (5), parafoveal telangiectasia (2), or chorioretinitis (3). In two patients an idiopathic origin of the CNV was presumed. Another 21 patients showed a juxtafoveal classic membrane (12) or a recurrence after laser photocoagulation (9). Visual acuity was evaluated, also in correlation to age and CNV size. Continuous follow-up of CNV and leakage size was performed with fluorescein angiography. RESULTS: Overall 85% of the patients showed visual stabilization or improvement during an average observation period of 10.5 months. A correlation was found between age and visual improvement favoring younger patients; however, no correlation was found between visual improvement and CNV size. CNV and leakage size decreased moderately over the 9-month follow-up from 1.81/3.45 mm(2) to 1.61/2.60 mm(2). CONCLUSION: PDT shows efficacy for classic choroidal neovascularization beyond the common indication. PMID- 12748805 TI - [Impairment of safety in navigation caused by alcohol: impact on visual function]. AB - BACKGROUND: So far in Germany, no legally binding standards for blood alcohol concentration exist that prove an impairment of navigability. The aim of our interdisciplinary project was to obtain data in order to identify critical blood alcohol limits. In this context the visual system seems to be of decisive importance. MATERIALS AND METHODS: 21 professional skippers underwent realistic navigational demands soberly and alcoholized in a sea traffic simulator. The following parameters were considered: visual acuity, stereopsis, color vision, and accommodation. RESULTS: Under the influence of alcohol (average blood alcohol concentration: 1.08 per thousand ) each skipper considered himself to be completely capable of navigating. While simulations were running, all of the skippers made nautical mistakes or underestimated dangerous situations. Severe impairment in visual acuity or binocular function were not observed. Accommodation decreased by an average of 18% ( p=0.0001). In the test of color vision skippers made more mistakes ( p=0.017) and the time needed for this test was prolonged ( p=0.004). CONCLUSIONS: Changes in visual function as well as vegetative and psychological reactions could be the cause of mistakes and alcohol should therefore be regarded as a severe risk factor for security in sea navigation. PMID- 12748806 TI - [Impairment of safety in navigation caused by alcohol: pupillography and measurement of pupil movements]. AB - OBJECTIVE: To evaluate data in an interdisciplinary project (ophthalmology, forensic medicine, internal medicine, psychology,and nautical science) in order to identify critical blood alcohol limits in sea navigation. METHODS: A sea traffic simulator was employed for realistic nautical demands on 21 professional experienced skippers under sober and alcoholized conditions (target blood alcohol concentration: 1.0 per thousand ). After simulated navigation, pupil light reflex, spontaneous pupil movements,nystagmus,and saccades were evaluated by pupillography. Modification of the pupillograph enabled us also to measure optokinetic nystagmus. RESULTS: Evaluation of the pupil light reflex revealed obvious changes in the extent of relative contraction and in redilatation time under the influence of alcohol. Diminished vigilance could be observed in all of the skippers when optokinetic nystagmus was tested. CONCLUSION: The pupillograph represents a suitable device for measuring functions of the visual and vegetative systems. Thus, the impact of these functions on nautical capability can be demonstrated. If further investigations such as ophthalmological, medical, psychological, and nautical evaluations are taken into account, it could be determined that blood alcohol levels of 1.0 per thousand may exclude safe navigation. PMID- 12748807 TI - [Vigabatrin-associated bilateral simple optic nerve atrophy with visual field constriction. A case report and a survey of the literature]. AB - CASE REPORT: A 70-year-old male with optic disc pallor and visual field constriction up to 35 degrees bilaterally was previously treated with vigabatrin. The cumulative dosage was 3.7 kg. The b-wave was reduced in photopic and scotopic electroretinography and the patient had delayed visual evoked potentials. The retinal nerve fiber thickness was pathologically reduced in the Heidelberg retinal tomography (HRT) and the retinal nerve fiber layer analyzer (GDx). CONCLUSION: Vigabatrin is an antiepileptic drug and may increase the intraretinal GABA level due to an inhibition of the enzyme GABA-transaminase and may induce an ascending simple optic nerve atrophy. Vigabatrin-associated visual field defects may be irreversible. A visual field control is recommended before starting the treatment with vigabatrin and at regular 6-month intervals thereafter. In cases of visual field constriction, the therapy should be stopped. If the cumulative dosage is more than 3 kg of vigabatrin, the visual field controls should be performed more frequently because of the dosage-toxicity relationship. PMID- 12748808 TI - [Nd-YAG laser iridotomy in pigment dispersion syndrome and pigment dispersion glaucoma]. AB - Pigment dispersion syndrome is characterized by iris transillumination defects, Krukenberg spindels, and dense trabecular pigmentation. Additional features are bilaterality, myopia, concavity of the peripheral iris, a higher incidence in men than in woman,and young age of onset. A mechanism of reverse pupillary block causes iridozonular friction. Laser iridotomy is recommended as treatment to prevent further pigment dispersion and pigmentary glaucoma. PMID- 12748809 TI - [Basic methods of strabismology]. AB - Basic examination techniques for manifest and latent strabismus as well as for paralytic strabismus are discussed. Usually the medical history gives the first clues for the form of strabismus. The examination starts with an evaluation of the head posture. The objective angle is estimated through corneal reflections and the corrective saccade (cover test). Uncover and cover testing allows the differentiation between manifest and latent strabismus. The Lang test, Bagolini's test and the light red glass test provide information regarding quality of binocularity (abnormal and normal retinal correspondence). Estimation of changes in the angle of strabismus in the diagnostic gaze positions by corneal reflections, cover test and the limbus test of Kestenbaum is complemented by testing of the visual acuity and fixation. Without evaluation of fixation neither the cover test nor tests for binocularity can be interpreted adequately. PMID- 12748810 TI - Urgent thoracic aortal dissection and aneurysm: treatment with stent-graft implantation in an angiographic suite. AB - The aim of this study was to evaluate the feasibility of endoluminal stent-graft placement in an angiographic suite for the treatment of emergent type-B aortic dissections and ruptured thoracic aortal aneurysms. Twenty-six patients with either urgent type-B dissection (n=8) or aneurysms (n=18) of the descending thoracic aorta were chosen for stent-graft implantation. All patients received a multidetector-row CT angiography of the whole aorta and pelvic arteries prior to stent-graft implantation. All procedures were performed in a fully equipped digital subtraction angiography (DSA) suite under general anesthesia. In 20 patients Talent LPS tube grafts and in 4 patients an Excluder graft were used. Access was achieved via surgical cut-down in the left (n=7) or right (n=19) groin. Sealing was successful in 24 patients. The proximal covered portion of the stent graft was placed across the left subclavian artery in 2 patients. Procedural success was achieved in 23 of 24 patients. One patient required a second stent-graft placement before the aneurysm was sealed. One patient with an acute perforation of the descending aorta died due to cardiac failure prior to stent-graft implantation. In 1 patient stent-graft delivery failed due to severe calcification of both common iliac arteries. Endoluminal treatment of both urgent type-B aortic dissections and thoracic aortal aneurysms with stent graft is an attractive alternative treatment to surgical repair. The placement of stent grafts in an angiographic suite is a safe and feasible method with good clinical effectiveness and, so far, good clinical outcome. PMID- 12748811 TI - Unstable pelvic insufficiency fracture in a patient with rheumatoid arthritis. AB - The occurrence of pelvic insufficiency fractures in patients with rheumatoid arthritis has not previously been well emphasized. These fractures are difficult to detect clinically, and appropriate radiological investigation is necessary for diagnosis. A 72-year-old woman with rheumatoid arthritis presented with severe left groin pain. Pelvic radiographs showed parasymphyseal fractures, and marked instability of these fractures was observed at the follow-up 2 weeks later. Computed tomographic scan of the sacrum showed a widened linear fracture gap in the left sacral ala. Because the patient's pain was so severe that she could not change position, external fixation was performed to achieve rapid pain relief and early mobilization. Although most patients with these fractures respond well to simple conservative treatments, parasymphyseal fractures combined with sacral fractures may cause disruption of the pelvic ring and occasionally need operative management. PMID- 12748812 TI - The Nag1 N-acetylglucosaminidase of Trichoderma atroviride is essential for chitinase induction by chitin and of major relevance to biocontrol. AB - The nag1 gene of the mycoparasitic fungus Trichoderma atroviride encodes a 73-kDa N-acetyl-beta- d-glucosaminidase, which is secreted into the medium and partially bound to the cell wall. To elucidate the role of this enzyme in chitinase induction and biocontrol, a nag1-disruption mutant was prepared. It displayed only 4% of the original N-acetyl-beta- d-glucosaminidase activity, indicating that the nag1 gene product accounts for the majority of this activity in T. atroviride. The nag1-disruption strain was indistinguishable from the parent strain in growth and morphology, but exhibited delayed autolysis. Northern analysis showed that colloidal chitin disruption does not induce ech42 gene transcription in the nag1-disruption strain. Enzyme activities capable of hydrolysing p-nitrophenyl- N, N'-diacetylchitobioside and p-nitrophenyl- N, N' diacetylchitotriose were also absent from the nag1-disruption strain under the same conditions. Retransformation of the T. atroviride nag1-disruption strain with the nag1 gene essentially led to the parent-type behaviour in all these experiments. However, addition of N-acetyl-beta- d-glucosaminidase to the medium of the nag1-disruption strain did not rescue the mutant phenotype. The disruption nag1 strain showed 30% reduced ability to protect beans against infection by Rhizoctonia solani and Sclerotinia sclerotiorum. The data indicate that nag1 is essential for triggering chitinase gene expression in T. atroviride and that its functional impairment reduces biocontrol by T. atroviride by a significant extent. PMID- 12748813 TI - Polyadenylation of rRNA- and tRNA-based yeast transcripts cleaved by internal ribozyme activity. AB - Polyadenylation, an important step in 3' end-processing of mRNA in eukaryotes, results in a poly(A) tail that ensures RNA transport into the cytoplasm and subsequent translation. Addition of a poly(A) tail is restricted to transcripts that are synthesized by RNA polymerase II. Here, we demonstrate that the 3' ends of yeast transcripts based on rRNA and tRNA, respectively, can be polyadenylated in vivo. The transcripts were modified by insertion of a self-cleaving hammerhead ribozyme sequence in the corresponding gene. Both the rDNA-based transcript and the tRNA transcript were cleaved efficiently by the hammerhead ribozyme, resulting in two stable cleavage products. The 5' cleavage product was found to be polyadenylated in both cases. This demonstrates that, in yeast, transcripts that are usually synthesized by RNA polymerase I or III can be polyadenylated if the 3' end of the transcript has been generated independently by a ribozyme. PMID- 12748814 TI - Topographical anatomy on the communicating branch between the spinal accessory nerve and the anterior root of the first cervical nerve. AB - The aim of this study was to clarify the morphological characteristics of the communicating branch between the spinal accessory nerve and the anterior root of the first cervical nerve (McKenzie branch), a structure severed during intradural selective rhizotomy for treatment of spasmodic torticollis. The right and left sides of 50 upper cervical segments of spinal cords were dissected and examined under a surgical microscope. Seven of the total 100 samples were found to have the McKenzie branch. Two types of McKenzie branch, which run close to the vertebral artery, were also identified, which might be helpful for improving the surgical results of intradural selective rhizotomy. PMID- 12748815 TI - Measurements of the lumbar pedicles in the Eastern Anatolian population. AB - Pedicle screw fixation of the spine has become one of the most stable and versatile methods of spine fixation, and knowledge of pedicle morphology is crucial for the safe application of these systems. We undertook this study because only a few reports have investigated Eastern populations. Lumbar pedicle anatomy, i.e., pedicle width (PW) and pedicle height (PH), transverse and sagittal pedicle angles (TPA, SPA), and pedicle length (PL), were assessed in the following two groups: (1) computed tomography scans of 29 normal adults, and (2) 16 dried lumbar spines obtained from the Anatomy Department. Interpedicular distance was different in each group. PW ranged from 4 mm to 14 mm. In both groups, the narrowest PH was 8.2 mm, the widest 19.7 mm. TPA ranged from 6(o) to 19(o) and increased from L1 to L5. In the sagittal plane, the pedicles angled caudally at L5. PL was longest at L1 and shortest at L5. In conclusion, pedicle dimensions and angles may show individual and structural differences. Our data were not significantly different from previous reported data. A detailed knowledge of these relationships is important for any surgery involving screw purchase via a pedicle, to prevent screw cutout and failure of fixation or neurological injury. Selection of the proper diameter of screw is an important issue for safe placement. Knowledge of the pedicle axis length is essential in choosing screw lengths but should always be checked intraoperatively with fluoroscopic control during screw insertion. PMID- 12748816 TI - The anatomy of the carotico-clinoid foramen and its relation with the internal carotid artery. AB - The carotico-clinoid foramen is the result of ossification either of the carotico clinoid ligament or of a dural fold extending between the anterior and middle clinoid processes of the sphenoid bone. It is anatomically important due to its relations with the cavernous sinus and its content, sphenoid sinus and pituitary gland. In this study the ossification state of the carotico-clinoid ligament, the diameter of the internal carotid artery and the carotico-clinoid foramen has been studied on 50 autopsy cases. Of the 100 carotico-clinoid foramina examined, in 27 sides (15 right, 12 left) the carotico-clinoid ligament was completely ossified, in 18 sides (9 right, 9 left) the carotico-clinoid ligament was incompletely ossified and in 55 sides (26 right, 29 left) it was a ligamentous structure. The correlation of the dimensions of the carotico-clinoid foramen and the internal carotid artery showed no statistical significance, except between the carotico clinoid foramen with a fibrous carotico-clinoid ligament and the internal carotid artery on the right side (p=0.007, r=0.51). The existence of a bony carotico clinoid foramen may cause compression, tightening or stretching of the internal carotid artery. Further, removing the anterior clinoid process is an important step in regional surgery; the presence of a bony carotico-clinoid foramen may have high risk. Therefore, detailed knowledge of the type of ossification between the anterior and middle clinoid processes can be necessary to increase the success of regional surgery. PMID- 12748828 TI - Magnetic resonance imaging of the growth plate in late-onset tibia vara. AB - We used Magnetic resonance imaging (MRI) in five patients (six knees), mean age 13.2 (12-15) years, with late-onset tibia vara (Blount's disease), to study the growth plate and its abnormalities. The MRI study was classified for severity of disease and compared with a radiographic classification. In severely involved knees, MRI indicated severe growth-plate changes on both sides of the knee joint. Widening in the entire proximal tibial growth plate, involvement of the distal femoral growth plate, as well as cartilage invaginations into the metaphyses, were constant findings. Three knees were treated operatively with oblique tibial osteotomy and three with lateral hemiepiphysiodesis. Two severely involved patients treated initially with hemiepiphysiodesis required additional surgery. The three patients with mild disease treated with tibial osteotomy had good clinical and functional results. This study suggests that extensive growth-plate changes in severe, late-onset tibia vara preclude successful treatment by tibial hemiepiphysiodesis. In addition, oblique osteotomy, which was successful in mild cases, was problematic in severe cases. PMID- 12748829 TI - PET and restaging of malignant lymphoma including residual masses and relapse. AB - Differentiation of post-therapeutic scar tissue from active lymphoma is unsatisfactory when using only morphological imaging approaches. Positron emission tomography (PET) with fluorine-18 fluorodeoxyglucose (FDG) may provide superior clinical information by enabling biochemical tissue characterisation, with enhanced FDG uptake in viable post-therapeutic lymphoma masses and very low uptake in indolent fibrotic tissue. With this in mind, 15 recently published studies reporting the results of differentiation of viable lymphoma from scar tissue in 723 patients were analysed. Sensitivity of FDG-PET for detection of active disease was 71-100%, and the specificity was 69-100%. Accordingly, FDG-PET had a high negative predictive value of 80-100%. In contrast, the specificity and positive predictive value (PPV) of computed tomography were low (4-31% and 19 60%, respectively, except in one study in which the PPV was 82%). Residual masses that were positive on PET were associated with a progression-free survival of 0 40%. Although not perfect, the biochemical approach for characterisation of residual masses in lymphoma with FDG-PET at present seems the most accurate way to differentiate scar tissue from viable residual lymphoma. PMID- 12748830 TI - Impact of technology on the utilisation of positron emission tomography in lymphoma: current and future perspectives. AB - Positron emission tomography (PET) has now gained a place in the management of patients with cancer, including those with Hodgkin's disease and non-Hodgkin's lymphoma. Restaging studies and those addressing the monitoring of response to treatment are especially in focus. Most of the knowledge gained has been achieved with dedicated BGO-based PET technology, but there are a number of developments that will impact on the use of this metabolic imaging technique in the investigation of patients with lymphoma. The challenges ahead are determined by the need for high-quality whole-body imaging associated with increased patient throughput and the need to investigate the role of new labelled ligands. The latter are likely to yield new insights into tumour cell characterisation, tumour behaviour and tumour outcome assessment. The study of new radiolabelled ligands will impose further demands for rapid dynamic data acquisition and accurate tracer quantification. Current and future developments in PET technology range from the use of new detector materials to different detector geometries and data acquisition modes. The search for alternatives to BGO scintillation materials for PET has led to the development of PET instruments utilising new crystals such as LSO and GSO. The use of these new detectors and the increased sensitivity achieved with 3D data acquisitions represent the most significant current developments in the field. With the increasing demands imposed on the clinical utilisation of PET, issues such as study cost and patient throughput will emerge as significant future factors. As a consequence, low-cost units are being offered by the manufacturers through the utilisation of gamma camera-based SPET systems for PET coincidence imaging. Unfortunately, clinical studies in lymphoma and other cancers have already demonstrated the limitations of this technology, with 20% of lesions <15 mm in size escaping detection. On the other hand, the recent development of combined PET/CT devices attempts to address the lack of anatomical information inherent with PET images, taking advantage of further improvement in patient throughput and hence cost-effectiveness. Preliminary studies using this multimodality imaging approach have already demonstrated the potential of the technique. Although the potential exists, certain technical issues with PET/CT require refinement of the methodology. Such issues include organ movement (such as respiratory motion), which strongly influences the image fusion of a rapidly acquired CT scan with the slower acquisition of a PET dataset, and the derivation of CT-based attenuation coefficients in the presence of contrast agents or metallic implants. The application of the technology for radiotherapy planning also poses a number of associated challenges. Finally, the development of dedicated PET systems based on planar detector arrangements with new detector components has the potential to improve clinical throughput by over 100%, but clinical trials using such systems have still to be carried out in order to establish the associated whole-body image quality. PMID- 12748831 TI - Limitations of PET for imaging lymphoma. AB - The uptake of fluorine-18 fluorodeoxyglucose (FDG) is increased in processes with enhanced glycolysis, including malignancy. It is this property of FDG which is exploited in positron emission tomography (PET) imaging for lymphoma. FDG, whilst a good oncology tracer, is not perfect and there are limitations to its use. FDG may have low uptake in some types of lymphoma, predominantly low-grade lymphomas. High physiological uptake may occur within the bowel, urinary tract, muscle, salivary glands and lymphoid tissue. FDG is not specific for malignancy and increased uptake occurs in benign conditions with increased glycolysis such as infection, inflammation and granulomatous disease. Benign conditions usually have lower uptake than malignancy but there is overlap. These limitations of FDG mean that tumour may be 'missed', 'masked' or 'mimicked' by other pathology. These limitations are described in this article and methods to circumvent them where possible are discussed. These include performing baseline scans at presentation with lymphoma for comparison with post-treatment scans, simple manoeuvres to reduce physiological uptake such as administration of frusemide and diazepam and remaining alert to the possibility of alternative pathology in immunosuppressed patients. Patients with disease secondary to human immunodeficiency virus are a particular challenge in this regard as they often have dual or multiple pathology. One of the most important skills in PET reporting may be to recognise its limitations and be clear when a definitive answer cannot be given to the referring clinician's question. This may require using PET to direct the clinician to biopsy the site most likely to yield the correct diagnosis. PMID- 12748832 TI - Role of ventilation scintigraphy in diagnosis of acute pulmonary embolism: an evaluation using artificial neural networks. AB - The purpose of this study was to assess the value of the ventilation study in the diagnosis of acute pulmonary embolism using a new automated method. Either perfusion scintigrams alone or two different combinations of ventilation/perfusion scintigrams were used as the only source of information regarding pulmonary embolism. A completely automated method based on computerised image processing and artificial neural networks was used for the interpretation. Three artificial neural networks were trained for the diagnosis of pulmonary embolism. Each network was trained with 18 automatically obtained features. Three different sets of features originating from three sets of scintigrams were used. One network was trained using features obtained from each set of perfusion scintigrams, including six projections. The second network was trained using features from each set of (joint) ventilation and perfusion studies in six projections. A third network was trained using features from the perfusion study in six projections combined with a single ventilation image from the posterior view. A total of 1,087 scintigrams from patients with suspected pulmonary embolism were used for network training. The test group consisted of 102 patients who had undergone both scintigraphy and pulmonary angiography. Performances in the test group were measured as area under the receiver operation characteristic curve. The performance of the neural network in interpreting perfusion scintigrams alone was 0.79 (95% confidence limits 0.71-0.86). When one ventilation image (posterior view) was added to the perfusion study, the performance was 0.84 (0.77-0.90). This increase was statistically significant ( P=0.022). The performance increased to 0.87 (0.81-0.93) when all perfusion and ventilation images were used, and the increase in performance from 0.79 to 0.87 was also statistically significant ( P=0.016). The automated method presented here for the interpretation of lung scintigrams shows a significant increase in performance when one or all ventilation images are added to the six perfusion images. Thus, the ventilation study has a significant role in the diagnosis of acute lung embolism. PMID- 12748833 TI - Preoperative mapping of cortical language areas in adult brain tumour patients using PET and individual non-normalised SPM analyses. AB - In patients scheduled for the resection of perisylvian brain tumours, knowledge of the cortical topography of language functions is crucial in order to avoid neurological deficits. We investigated the applicability of statistical parametric mapping (SPM) without stereotactic normalisation for individual preoperative language function brain mapping using positron emission tomography (PET). Seven right-handed adult patients with left-sided brain tumours (six frontal and one temporal) underwent 12 oxygen-15 labelled water PET scans during overt verb generation and rest. Individual activation maps were calculated for P<0.005 and P<0.001 without anatomical normalisation and overlaid onto the individuals' magnetic resonance images for preoperative planning. Activations corresponding to Broca's and Wernicke's areas were found in five and six cases, respectively, for P<0.005 and in three and six cases, respectively, for P<0.001. One patient with a glioma located in the classical Broca's area without aphasic symptoms presented an activation of the adjacent inferior frontal cortex and of a right-sided area homologous to Broca's area. Four additional patients with left frontal tumours also presented activations of the right-sided Broca's homologue; two of these showed aphasic symptoms and two only a weak or no activation of Broca's area. Other frequently observed activations included bilaterally the superior temporal gyri, prefrontal cortices, anterior insulae, motor areas and the cerebellum. The middle and inferior temporal gyri were activated predominantly on the left. An SPM group analysis ( P<0.05, corrected) in patients with left frontal tumours confirmed the activation pattern shown by the individual analyses. We conclude that SPM analyses without stereotactic normalisation offer a promising alternative for analysing individual preoperative language function brain mapping studies. The observed right frontal activations agree with proposed reorganisation processes, but they may also reflect an unspecific recruitment of the right-sided Broca's homologue in the effort to perform the task. PMID- 12748834 TI - Renal cell carcinoma in long-term survivors of advanced stage neuroblastoma in early childhood. AB - BACKGROUND: Renal cell carcinoma (RCC) is rare in children and comprises only 1 3% of all pediatric primary renal tumors. Recently, several case reports have described RCC developing in patients previously treated for advanced stage neuroblastoma (NB). METHODS AND RESULTS: Our experience with four patients treated for advanced stage NB during early childhood who developed RCC later in life are added to 14 others in the literature. CONCLUSION: These patients and our review of the literature suggest an association between RCC and NB that warrants further study. PMID- 12748835 TI - A flow rate cut-off value as a criterion for the accurate non-invasive measurement of bladder pressure using a condom-type catheter. AB - We developed a condom-type catheter to non-invasively measure the bladder pressure during interruption of the flow rate. The aim of the present study was to establish a minimum flow rate value at which a reliable bladder pressure measurement can be made with this catheter. We reanalysed data from 43 patients who completed a pressure-flow study and a non-invasive test. The patients voided without straining. During the test, we simultaneously measured the bladder pressure (invasively) and the condom pressure (non-invasively). The pressure increase in the condom after interruption of the flow rate was analysed in 40 of the 43 patients. A plot of the difference between the bladder pressure and the maximum condom pressure as a function of the flow rate revealed that in 70% of the patients who voided with a maximum flow rate exceeding 5.4 ml/s, the condom pressure accurately reflected the bladder pressure (+/-14 cmH2O). We conclude that to accurately and non-invasively measure the bladder pressure with a condom type catheter, the maximum flow rate should exceed 5.4 ml/s. PMID- 12748836 TI - Cystine crystal volume determination: a useful tool in the management of cystinuric patients. AB - We prospectively determined cystine crystal volume (Vcys) in urine specimens from all consecutive patients with cystine urolithiasis followed at our institution over the past decade, in order to assess its predictive value as to the risk of recurrent cystine stone formation. A total of 57 patients (29 males, 28 females) with homozygous cystinuria entered in the study between January 1990 and December 2000, including 15 children aged less than 15 years and 42 patients aged 15 years or more. The clinical and radiological course was followed until December 2001, for a total of 243 patient-years of follow-up. From study entry until the end of follow-up, we serially examined first voided morning urine specimens in all patients, with determination of the number of cystine crystals per mm3, and the average size of crystals, thus allowing us to calculate Vcys using a simple formula based on crystal geometry. Recurrence was diagnosed on the basis of serial radiographic examinations using X-rays and echography. Overall, cystine crystals were present in 179 (39%) of the 460 examined urine specimens. Cystine crystalluria was significantly more frequent among the 27 patients who developed new cystine stones (SF) than in the other 30 who remained stone-free (63.3 vs 25.5% of samples, P<0.001). The presence of crystals in > or =50% of serially examined urine samples was more frequently found in patients with recurrent stone formation than in non-recurrent patients (24/27 vs 2/30, P<0.001). The average Vcys value was significantly higher in recurrent SF than in stone-free patients (8,173 +/- 1,544 vs 233 +/- 150 micro3/mm3, P<0.001) and there was no overlap in the individual values of recurrent vs stone-free patients. A Vcys value > or =3,000 micro3/mm3 was observed at least once prior to each of the 63 stone recurrences observed in 27 patients (2.3 per patient on the average). In addition, Vcys reflected the efficacy of treatment, with Vcys mean values of 12,097 +/- 3,214 micro3/mm3 at baseline, falling to 2,648 +/- 658 micro3/mm3 on basic therapy (hyperdiuresis plus alkalinization) alone, 1,141 +/- 522 micro3/mm3 on tiopronin therapy (median dose 1,000 mg/day) and 791 +/- 390 micro3/mm3 on D penicillamine therapy (median dose 900 mg/day) whereas captopril had no effect (5,114 +/- 2,128 micro3/mm3). Based on the results of the present study, cystine crystalluria appears to accurately reflect active stone formation in cystinuric patients. Determination of total Vcys provides a simple, cheap and accurate means of predicting the risk of cystine stone recurrence with a Vcys value > or =3000 micro3/mm3 as the threshold risk value. We propose that serial Vcys determination be performed simultaneously with the measurement of urine pH and specific gravity to optimally monitor the medical treatment of cystine patients. PMID- 12748837 TI - Obstacle avoidance during human walking: H-reflex modulation during motor learning. AB - The goal of this study was to investigate changes of H-reflex amplitudes during a motor learning task. Subjects with reduced vision were instructed to step over an obstacle on a treadmill as low as possible, while the soleus H-reflex was elicited. Acoustic warning and feedback signals about performance were provided. Performance improvement was associated with a decrease of muscle activity, needed to step over the obstacle (rectus femoris, biceps femoris, tibialis anterior and gastrocnemius medialis muscles), and of foot clearance, while joint angle trajectories from knee and ankle became more stable. The experiment consisted of five runs, three with normal treadmill walking and two with randomly stepping over the obstacle (100 times). H-reflexes were elicited at early and late stance phase before stepping over the obstacle. H/M ratio, latency and duration were determined. The values of these measures were calculated for the onset and end of a run and their course over time was evaluated using a correlation coefficient. The largest adaptations with a significant increase of reflex amplitude occurred during the first obstacle run. This increase lasted only briefly and the reflex amplitudes decreased to their previous values. During the later obstacle run, no H-reflex modulation occurred. It is concluded that a motor learning task causes adaptational effects not only on performance, but also on H-reflex responses. The results indicate that most of the modulation of H-reflexes is probably due to supraspinal influences on reflex transmission. The observations made are probably less specific for this motor task (stepping over the obstacle), but rather associated with the increased attention required by the motor learning task during the first obstacle run. PMID- 12748839 TI - Preliminary assessment of arbuscular mycorrhizal fungal diversity and community structure in an urban ecosystem. AB - Arbuscular mycorrhizal fungal (AMF) species richness, composition, spore density and diversity indices were evaluated in the Phoenix metropolitan area, Arizona, USA at 20 sampling sites selected to represent the four predominant land-use types found in the greater urban area: urban-residential, urban non-residential, agriculture and desert. AMF spores were extracted and identified from soil samples and from trap cultures established using soil collected at each site. Data were analyzed according to land use, land-use history, soil chemistry and vegetation characteristics at each site. Current agricultural sites were associated with decreased spore densities and historically agricultural sites with decreased species richness. Overall species composition was similar to that previously reported for the Sonoran desert, but composition at each sampling site was influenced by the vegetation from which samples were collected. Sites with the highest degrees of similarity in AMF species composition were also similar to each other in native plants or land use. Conversely, sites with the lowest similarity in AMF composition were those from which the majority of samples were collected from non-mycorrhizal plants, predominately ectomycorrhizal plants or bare soil. Spores of Glomus microggregatum were most abundant in urban sites, while those of G. eburneum were most abundant in desert and agricultural sites. Further studies are needed to determine the functional implications of shifts in AMF communities in urban ecosystems, including effects on plant primary productivity. PMID- 12748838 TI - Connectivity of the goldfish optic tectum with the mesencephalic and rhombencephalic reticular formation. AB - The optic tectum of goldfish, as in other vertebrates, plays a major role in the generation of orienting movements, including eye saccades. To perform these movements, the optic tectum sends a motor command through the mesencephalic and rhombencephalic reticular formation, to the extraocular motoneurons. Furthermore, the tectal command is adjusted by a feedback signal arising from the reticular targets. Since the features of the motor command change with respect to the tectal site, the present work was devoted to determining, quantitatively, the particular reciprocal connectivity between the reticular regions and tectal sites having different motor properties. With this aim, the bidirectional tracer, biotin dextran amine, was injected into anteromedial tectal sites, where eye movements with small horizontal and large vertical components were evoked, or into posteromedial tectal sites, where eye movements with large horizontal and small vertical components were evoked. Labeled boutons and somas were then located and counted in the reticular formation. Both were more numerous in the mesencephalon than in the rhombencephalon, and ipsilaterally than contralaterally, with respect to the injection site. Furthermore, the somas showed a tendency to be located in the area containing the most dense labeling of synaptic endings. In addition, labeled boutons were often observed in close association with retrogradely stained neurons, suggesting the presence of a tectoreticular feedback circuit. Following the injection in the anteromedial tectum, most of the boutons and labeled neurons were found in the reticular formation rostral to the oculomotor nucleus. Conversely, following the injection in the posteromedial tectum, most of the boutons and neurons were also located in the caudal mesencephalic reticular formation. Finally, boutons and neurons were found in the rhombencephalic reticular formation surrounding the abducens nucleus. They were more numerous following the injection in the posteromedial tectum. These results demonstrate characteristic patterns of reciprocal connectivity between physiologically different tectal sites and the mesencephalic and rhombencephalic reticular formation. These patterns are discussed in the framework of the neural substratum that underlies the codification of orienting movements in goldfish. PMID- 12748840 TI - Arbuscular mycorrhiza colonization and development at suboptimal root zone temperature. AB - Temperature has a strong influence on the activity of living organisms. This study, involving two indoor experiments, evaluated the effects of root zone temperature (10, 15 and 23 degrees C) on the formation and development of arbuscular mycorrhizae (AM). In the first trial, greenhouse-grown sorghum [ Sorghum bicolor (L.) Moench] was either colonized by Glomus intraradices Schenck & Smith or left non-mycorrhizal. Root length, root and shoot weight and root colonization were measured after 5, 10 and 15 weeks of plant growth. Although suboptimal root zone temperatures reduced growth in both mycorrhizal and non mycorrhizal plants, mycorrhizal plants were larger than non-mycorrhizal plants after 15 weeks at 15 and 23 degrees C. At suboptimal root zone temperatures, mycorrhizal inoculation sometimes slightly reduced root development. AM colonization was more affected than root growth at suboptimal root zone temperatures. Colonization was markedly reduced at 15 degrees C compared with 23 degrees C, and almost completely inhibited at 10 degrees C. The second experiment was conducted in vitro using transformed carrot ( Daucus carota L.) roots supporting G. intraradices. Mycelium length and spore number were measured weekly for 15 weeks. Spore metabolic activity (iodonitrotetrazolium reduction), root length and percentage root colonization were measured after 15 weeks. G. intraradices sporulation was reduced at temperatures below 23 degrees C, while spore metabolic activity was significantly reduced only at 10 degrees C. Root length and in particular percentage colonization were decreased at suboptimal temperatures. A negative interaction between AM hyphal growth and root growth resulting in reduced probability of contact at suboptimal root zone temperatures is proposed to explain the greater reduction observed in root colonization than in root and hyphal growth. PMID- 12748841 TI - Effect of air temperature on forecasting the start of the Betula pollen season at two contrasting sites in the south of Europe (1995-2001). AB - In order to survive periods of adverse cold climatic conditions, plant requirements are satisfied by means of physiological adaptations to prevent cells from freezing. Thus, the growth of woody plants in temperate regions slows down and they enter into a physiological state called dormancy. In order to identify the chilling and heat requirements to overcome the dormancy period of Betula in the south of Europe, a comparative study was carried out with aerobiological pollen data of a 7-year (1995-2001) period in Vigo (Spain) and Perugia (Italy). To satisfy chilling requirements, base temperatures of 7 degrees C and 5.75 degrees C showed a lower standard variation coefficient: 3.94% and 2.36% in Perugia and Vigo respectively. In the case of heat accumulation, the sum of mean temperatures in Perugia and the sum of maximum temperatures in Vigo were the parameters that showed a minor coefficient of variation (11.13% and 14.51% respectively). PMID- 12748842 TI - Face cooling by cold wind in walking subjects. AB - The effects of low to moderate wind speeds on face temperature, thermal and pain sensations while subjects walked on a treadmill during cold exposure were studied in eight healthy men. The purpose of the study was to evaluate further the risk of frostbite at different activity levels. The walking speed was 2.8 km h(-1) and two inclination levels were used, 0 degrees and 6 degrees. The subjects were exposed to -10 degrees C and 0, 1 or 5 m s(-1) wind for 60 min dressed in cold protective clothing with only the face unprotected. Results from previous experiments with the same subjects standing for 30 min were included in the analysis of the data. Each individual was exposed to all combinations of air velocity and activity level. The exposure to -10 degrees C and the highest wind speed used would carry no risk of frostbite according to the wind chill index. Cold lowered the skin temperature of the face significantly and wind further increased skin cooling. The activity level did not affect forehead and cheek temperatures, but the average nose skin temperature was higher and pain sensations were reduced at a higher work rate. The predicted risk of frostbite in the nose, based on average responses, would thus be less at a higher work rate. However, the results indicate that exercise does not necessarily protect all individuals from frostbite at moderate air speeds, since the nose skin temperature of 25% of the subjects dropped to 0 degrees C at 5 m s(-1) during both standing and walking. Thus the potential individual risk of frostbite in the nose is similar during light exercise and standing. Moreover, the risk of frostbite seems to be underestimated by the wind chill index under the conditions tested in this study. PMID- 12748843 TI - The impact of a small lake on heat stress in a Mediterranean urban park: the case of Tel Aviv, Israel. AB - Field observations of air and surface temperatures, relative humidity, solar radiation and wind were performed in the daytime hours of the warm season around a pond of 4 ha, located in Begin Park, in the city of Tel Aviv, Israel. Observations were carried out at screened meteorological stations on four randomly selected days, all associated with moderate heat stress. Two of them, one representing a warm and dry day, and other, representing a sultry day, are analyzed in detail. At the downwind side of the pond, lower temperatures, a higher relative humidity and a lower heat stress index were observed consistently when compared with stations located upwind of the pond. This effect is regarded here as the "lake effect". The fact that no significant change was noted in the water vapor pressure during most of the daytime hours indicates that the lake effect was related mainly to cooling rather than to moisture transport from the pond. A positive relationship was found between the lake effect and wind speed in both types of weather. The maximum effect of the wind's speed on the lake effect was observed at midday, at which time the temperature drop reached 1.6 degrees C, while the relative humidity rose by 6%. As a result, the heat stress index dropped by 0.8-1.1 degrees C. It is suggested that the temperature drop induced by the pond during the warmest hours of the day was mainly the result of a truncation of the sensible heat flux from the underlying surface when the air, which had previously passed over hot surfaces, swept over the relatively cool water. During the late afternoon and evening hours, when the water became warmer than the surrounding surfaces, latent heat cooling resulting from evaporation became the dominant source of the lake effect, and the lake effect resulted in increasing heat stress. It is concluded that even small bodies of water have a relieving effect on humans in the daytime hours, within the range of 40 m, under both dry and humid hot weather conditions. PMID- 12748844 TI - Effective chemical preservation of morphology of urinary erythrocytes. AB - Between 2 and 4 h after miction the morphology of urinary erythrocytes has changed, clouding the distinction between glomerular and non-glomerular bleeding in pediatric patients. Glomerular bleeding is characterized by microscopically visible alteration in urinary erythrocytes due to glomerular disease. Fixation by Cellfix, a formaldehyde-based fixative, allows the preservation of the morphology for at least 24 h and can be recommended for clinical practice. In our experience, thiomersal was not effective for preservation of the morphology of erythrocytes. PMID- 12748845 TI - Clinical quiz. Leptospirosis. PMID- 12748847 TI - Determination of intact splenic weight based on morcellated weight. AB - BACKGROUND: Comparisons of splenic size based on splenic weight are difficult after laparoscopic splenectomy, which results in a morcellated specimen. We report the results of a direct comparison between morcellated and intact splenic weights. METHODS: Porcine spleens were harvested via a midline laparotomy, and an intact splenic weight was obtained, which served as the control. The spleen then was placed into an impermeable retrieval bag and returned to the peritoneal cavity. A separate 10-mm incision was made and the spleen mechanically morcellated with a uterine forceps. This design most faithfully recreates the morcellation process during laparoscopic splenectomy in humans. The aggregate weight of the fragments was compared with intact splenic weight. RESULTS: Intact and morcellated weights were obtained from 58 porcine spleens. The mean intact splenic weight was 145 g, and the mean morcellated weight was 78 g. For a given morcellated weight achieved at laparoscopic splenectomy, an estimated intact weight can be determined by the following formula: intact weight (g) = morcellated weight (g) x 1.34 + 45. CONCLUSIONS: On the basis of our calculations, a normal spleen weighing 150 g would have a mean morcellated weight of 78 g, and splenomegaly (intact spleen weighing 250 g or more) would be defined by a morcellated weight exceeding 153 g. PMID- 12748848 TI - Molecular characterization and differentiation of opisthorchiid trematodes of the species Opisthorchis felineus (Rivolta, 1884) and Metorchis bilis (Braun, 1790) using polymerase chain reaction. AB - Adult specimens of the opisthorchiid liver fluke species Opisthorchis felineus and Metorchis bilis could be identified for the first time by molecular biological methods using species specific primers (OF and MB primers) in the polymerase chain reaction (PCR). The OF or MB primers were based on a part of the mitochondrial cytochrome c oxidase I gene. A specific product of approximately 200 bp could be amplified for O. felineus by means of the specific O. felineus primers. By contrast, the amplification of M. bilis DNA with MB primers produced a fragment of approximately 110 bp. A specificity of 100% could be demonstrated for both primer pairs. The sensitivities of the PCRs were 10 pg for the O. felineus DNA and 100 fg for the M. bilis DNA. PMID- 12748849 TI - No evidence of Wolbachia endosymbiosis with Loa loa and Mansonella perstans. AB - Endosymbiotic Wolbachia bacteria from different filarial species, including major pathogens of humans such as Wuchereria bancrofti, Brugia malayi and Onchocerca volvulus, seem to play an important role in the development, viability and fertility of these worms. Wolbachia trigger inflammatory host responses as well as adverse reactions against standard treatment regimens and are therefore under investigation as novel treatment targets. We investigated whether Wolbachia are also endosymbiotic in Loa loa and Mansonella perstans. In both male and female adult L. loa, we found no evidence of bacteria by light or transmission electron microscopy. Furthermore, Wolbachia-specific PCR was negative in both L. loa and M. perstans microfilariae. The absence of Wolbachia in both filarial species therefore discourages the use of antibiotics as an adjunct or alternative approach to current treatment concepts for both loiasis and mansonelliasis perstans. PMID- 12748850 TI - Growth-promoting effect of muscarinic acetylcholine receptors in colon cancer cells. AB - PURPOSE: G-protein-coupled receptors are known to mediate cell growth via divergent signaling pathways. It has been reported that colon cancer cells express muscarinic acetylcholine receptor (mAChR) although their functional role is largely unknown. The aim of this study is to elucidate possible mechanisms responsible for the growth-promoting effect of mAChRs in colon cancer cells by using colon cancer cell line T84. METHODS: Carbachol, a stable mAChR agonist, dose-dependently induced cell growth with a maximal effect observed at 100 microM, equipotent with 1 nM EGF. 4-DAMP, a specific antagonist of subtype 3 mAChR, inhibited the stimulatory effect by carbachol, suggesting that the growth promoting effect was receptor-mediated. Carbachol also dose-dependently stimulated extracellular signal-regulated protein kinase (ERK) activation. This effect was inhibited by PD98059, an inhibitor of extracellular signal-regulated protein kinase kinase, which also blocked carbachol activation of cell proliferation, indicating that the p21Ras-ERK pathway is an important signaling cascade in the mitogenic effect. To investigate how mAChR activated the p21Ras ERK pathway, transactivation of epidermal growth factor receptor (EGFR) was examined. RESULTS: Carbachol induced tyrosine phosphorylation of EGFR, which was abolished by an EGFR tyrosine kinase inhibitor AG1478. Transactivation by carbachol was also abrogated by a metalloproteinases (MMPs) inhibitor GM6001 or an EGFR-blocking antibody (LA-1), suggesting that binding of EGFR ligand(s) produced by MMPs may initiate transactivation in a manner dependent on EGFR tyrosine kinase. The tyrosine-phosphorylated EGFR was immunoprecipitated together with GRB2 and tyrosine-phosphorylated Shc, indicating that transactivated EGFR is able to generate downstream signals. AG 1478 and LA-1 inhibited carbachol stimulation of cell growth. CONCLUSIONS: Taken together, our results indicate that the growth-promoting effect of subtype 3 mAChR in colon cancer cells may depend on transactivated EGFR-ERK pathways. EGFR not only receives external stimuli but also serves as a scaffold for downstream signaling molecules. PMID- 12748851 TI - The incidence, prognosis, clinical and histological characteristics, treatment, and outcome of patients with bilateral germ cell testicular cancer in Hungary. AB - PURPOSE: To examine the incidence, prognosis, clinical and histological characteristics, treatment, and outcome of patients with bilateral testicular cancer in the referral center in Hungary, to determine which parameters might predict a second testicular tumor. METHODS: . Clinical parameters-such as time of original surgery, histology of primary tumor, extent of the disease, serum marker concentrations, history of testicular abnormalities, treatment, response to treatment, follow-up period, data on second carcinoma-of bilateral testicular tumors among the 2,386 patients with testicular cancer treated between November 1988 and November 1998 were analyzed. RESULTS: The incidence of patients with synchronous testicular tumor was 0.8% (19 of 2,386 patients). The clinical stages were 8 I/A, 5 I/B, 1 II/A, 2 II/B, 1 III/A, and 2 III/B. Median follow-up time was 93 months and the 5-year overall survival was 84%. The incidence of patients with metachronous testicular cancer (median age 28 years and 35 years at first and second tumor diagnosis) was 2.2% (53 of 2,386 patients) and the median time to second tumor was 76 months (range 18-203 months). The clinical stages at the first and second tumor diagnosis were: 14 I/A, 21 I/B, 15 II/A, 2 II/B, and 1 III/B, and 26 I/A, 16 I/B, 3 II/A, 1 II/B, 7 III/B, respectively. The median follow-up time was 42 months and the 5-year overall survival was 93%. In thirteen patients with metachronous cancers, two family histories of testicular cancer, five cases of undescended testicles, seven cases of testicular atrophy, and one case of azoospermia were detected. There was a non-significant trend to a longer cancer interval after chemotherapy and radiotherapy and a tendency to a greater incidence of asynchronous seminoma after chemotherapy. Clinical stage I tumors were more frequent in the surveyed group than among patients not followed up according to the institutional protocol ( P = 0.01), but the survival rate was good in both groups. Seminoma as a second tumor was diagnosed in an older age group (median 38 years, range 25-49 years) than nonseminoma (median 32 years, range 21-51 years, P < 0.045). The interval till the appearance of a metachronous testicular cancer depended on tumor histology: in seminoma cases it was longer than in nonseminoma cases (median time: 121 months versus 50 months, P = 0.002). CONCLUSIONS: The overall incidence of bilateral testicular cancer in the referral center in Hungary was 3%. We could not identify clinical factors which predicted a higher risk for metachronous testicular cancer. With regular follow-up the early diagnosis of second testicular tumors is probable; therefore education, self-examination of the remaining testicle, and long-term follow-up are important in early detection. PMID- 12748853 TI - A new case of mucopolysaccharidosis VII presenting as non immune hydrops fetalis. PMID- 12748852 TI - Megadose methylprednisolone for Kasabach-Merritt syndrome. PMID- 12748855 TI - Substrate specificity of the violaxanthin de-epoxidase of the primitive green alga Mantoniella squamata (Prasinophyceae). AB - The substrate specificity of the enzyme violaxanthin de-epoxidase (VDE) of the primitive green alga Mantoniella squamata (Prasinophyceae) was tested in in vitro enzyme assays employing the following xanthophyll mono-epoxides: antheraxanthin (Ax), diadinoxanthin (Ddx), lutein-epoxide (LE), cryptoxanthin-epoxide (CxE), 9- cis neoxanthin (cNx), all- trans neoxanthin (Nx), and xanthophyll di-epoxides: 9- cis violaxanthin (cVx), all- trans violaxanthin (Vx), cryptoxanthin-di-epoxide (CxDE). The data presented in this study show that the VDE of M. squamata not only exhibits a low affinity for the mono-epoxide Ax, as has been reported by R. Frommolt et al. (2001, Planta 213:446-456), but has a reduced substrate affinity for the mono-epoxides Ddx, LE, CxE, and Nx as well. On the other hand, xanthophylls with a second epoxy-group (Vx, CxDE) can be de-epoxidized with a higher efficiency. Such a preference for xanthophyll di-epoxides cannot be observed for the higher-plant VDE, where, in general, no marked differences in the pigment de-epoxidation rates between xanthophyll mono- and di-epoxides are visible. Despite this substantial difference between the VDEs of M. squamata and S. oleracea there are also features common to both enzymes. Neither VDE is able to convert xanthophylls with a 9- cis configuration in the acyclic polyene chain and both rely on substrates in the all- trans configuration. Both enzymes furthermore exhibit a dependence of enzyme activity on the polarity of the substrate. Highly polar (Nx) or non-polar (CxE) xanthophylls are de-epoxidized with greatly reduced rates in comparison to substrates with an intermediate polarity (Vx, Ax, LE, Ddx). This dependence on substrate polarity becomes more obvious when the higher-plant VDE is examined, as the substrate affinity of the VDE of M. squamata is more strongly influenced by the existence or absence of a second epoxy-group. In summary, the data presented in this study underline the fact that different VDEs, although in general catalyzing the same reaction sequence, are functionally diverse. PMID- 12748854 TI - Conserved expression of Hoxa1 in neurons at the ventral forebrain/midbrain boundary of vertebrates. AB - The previously described expression patterns of zebrafish and mouse Hoxa1 genes are seemingly very disparate, with mouse Hoxa1 expressed in the gastrula stage hindbrain and the orthologous zebrafish hoxa1a gene expressed in cell clusters within the ventral forebrain and midbrain. To investigate the evolution of Hox gene deployment within the vertebrate CNS, we have performed a comparative expression analysis of Hoxa1 orthologs in a range of vertebrate species, comprising representatives from the two major lineages of vertebrates (actinopterygians and sarcopterygians). We find that fore/midbrain expression of hoxa1a is conserved within the teleosts, as it is shared by the ostariophysan teleost zebrafish (Danio rerio) and the distantly related acanthopterygian teleost medaka (Oryzias latipes). Furthermore, we find that in addition to the described gastrula stage hindbrain expression of mouse Hoxa1, there is a previously unreported neurula stage expression domain, again located more anteriorly at the ventral fore/midbrain boundary. A two-phase expression profile in early hindbrain and later fore/midbrain is shared by the other tetrapod model organisms chick and Xenopus. We show that the anterior Hoxa1 expression domain is localized to the anterior terminus of the medial longitudinal fasciculus (MLF) in mouse, chick, and zebrafish. These findings suggest that anterior expression of Hoxa1 is a primitive characteristic that is shared by the two major vertebrate lineages. We conclude that Hox gene expression within the vertebrate CNS is not confined exclusively to the segmented hindbrain and spinal cord, but rather that a presumptive fore/midbrain expression domain arose early in vertebrate origins and has been conserved for at least 400 million years. PMID- 12748856 TI - (Patho)physiological implications of the novel epithelial Ca2+ channels TRPV5 and TRPV6. AB - The epithelial Ca(2+) channels TRPV5 and TRPV6 constitute the apical Ca(2+) entry mechanism in active Ca(2+) (re)absorption. These two members of the superfamily of transient receptor potential (TRP) channels were cloned from the vitamin-D responsive epithelia of kidney and small intestine and subsequently identified in other tissues such as bone, pancreas and prostate. These channels are regulated by vitamin D as exemplified in animal models of vitamin-D-deficiency rickets. In addition, the epithelial Ca(2+) channels might be involved in the multifactorial pathogenesis of disorders ranging from idiopathic hypercalciuria, stone disease and postmenopausal osteoporosis. This review highlights the emerging (patho)physiological implications of these epithelial Ca(2+) channels. PMID- 12748857 TI - Impaired PTH-induced endocytotic down-regulation of the renal type IIa Na+/Pi cotransporter in RAP-deficient mice with reduced megalin expression. AB - Inorganic phosphate (P(i)) reabsorption in the renal proximal tubule occurs mostly via the Na(+)/P(i) cotransporter type IIa (NaP(i)-IIa) located in the brush-border membrane (BBM) and is regulated, among other factors, by dietary P(i) intake and parathyroid hormone (PTH). The PTH-induced inhibition of P(i) reabsorption is mediated by endocytosis of Na/P(i)-IIa from the BBM and subsequent lysosomal degradation. Megalin is involved in receptor-mediated endocytosis of proteins from the urine in the renal proximal tubule. The recently identified receptor-associated protein (RAP) is a novel type of chaperone responsible for the intracellular transport of endocytotic receptors such as megalin. Gene disruption of RAP leads to a decrease of megalin in the BBM and to a disturbed proximal tubular endocytotic machinery. Here we investigated whether the distribution of NaP(i)-IIa and/or its regulation by dietary P(i) intake and PTH is affected in the proximal tubules of RAP-deficient mice as a model for megalin loss. In RAP-deficient mice megalin expression was strongly reduced and restricted to a subapical localization. NaP(i)-IIa protein distribution and abundance in the kidney was not altered. The localization and abundance of the NaP(i)-IIa interacting proteins MAP17, PDZK-1, D-AKAP2, and NHE-RF1 were also normal. Other transport proteins expressed in the BBM such as the Na(+)/H(+) exchanger NHE-3 and the Na(+)/sulphate cotransporter NaSi were normally expressed. In whole animals and in isolated fresh kidney slices the PTH-induced internalization of NaP(i)-IIa was strongly delayed in RAP-deficient mice. PTH receptor expression in the proximal tubule was not affected by the RAP knock-out. cAMP, cGMP or PKC activators induced internalization which was delayed in RAP deficient mice. In contrast, both wildtype and RAP-deficient mice were able to adapt to high-, normal, and low-P(i) diets appropriately as indicated by urinary P(i) excretion and NaP(i)-IIa protein abundance. PMID- 12748859 TI - Efflux and compartmentalization of zinc by members of the SLC30 family of solute carriers. AB - All of the members of this family are thought to facilitate zinc efflux from the cytoplasm either into various intracellular compartments (endosomes, secretory granules, synaptic vesicles, Golgi apparatus, or trans-Golgi network) or across the plasma membrane. Thus, these transporters are thought to help maintain zinc homeostasis and facilitate transport of zinc into specialized intracellular compartments. Counterparts of the SLC30 family are found in all organisms. Most of the members of this class are predicted to have 6 transmembrane domains with both N- and C-termini on the cytoplasmic side of the membrane. Expression of rodent Znt1, Znt2 or Znt4 cDNAs in mammalian cells can confer resistance to zinc toxicity. Loss of function of the mouse Znt1 is embryonic lethal, loss of mouse Znt3 prevents accumulation of zinc in synaptic vesicles, nonfunctional mouse Znt4 ( lethal milk) results in zinc-deficient milk, and Znt5-null mice display bone abnormalities and heart failure. No mutations in human counterparts of any of the members of the SLC30 family have been described. PMID- 12748860 TI - The SLC36 family: proton-coupled transporters for the absorption of selected amino acids from extracellular and intracellular proteolysis. AB - Whilst Na(+) has replaced H(+) as a major transport driving force at the plasma membrane of animal cells, the evolutionarily older H(+)-driven systems persist on endomembranes and at the plasma membrane of specialized cells. The first member of the SLC36 family, present in both intracellular and plasma membranes, was identified independently as a lysosomal amino acid transporter (LYAAT1) responsible for the export of lysosomal proteolysis products into the cytosol and as a proton/amino acid transporter (PAT1) responsible for the absorption of amino acids in the gut. In addition to LYAAT1/PAT1, the family comprises another characterized member, PAT2, and two orphan transporters. Both PAT1 and PAT2 mediate 1:1 symport of protons and small neutral amino acids such as glycine, alanine, and proline. Their mRNAs are broadly and differentially expressed in mammalian tissues. The PAT1 protein localizes to lysosomes in brain neurons, but is also found in the apical membrane of intestinal epithelial cells with a role in the absorption of amino acids from luminal protein digestion. In both cases, protons supplied by the lysosomal H(+)-ATPase or by the acidic microclimate of the brush border membrane drive transport of the amino acids into the cytosol. The subcellular localization and physiological role of PAT2 have still to be determined. SLC36 transporters are related distantly to other proton-coupled amino acid transporters, such as the vesicular neurotransmitter transporter VIAAT/VGAT (SLC32) and system N transporters (SLC38 family). PMID- 12748858 TI - The sodium/glucose cotransport family SLC5. AB - The sodium/glucose cotransporter family (SLCA5) has 220 or more members in animal and bacterial cells. There are 11 human genes expressed in tissues ranging from epithelia to the central nervous system. The functions of nine have been revealed by studies using heterologous expression systems: six are tightly coupled plasma membrane Na(+)/substrate cotransporters for solutes such as glucose, myo-inositol and iodide; one is a Na(+)/Cl(-)/choline cotransporter; one is an anion transporter; and another is a glucose-activated ion channel. The exon organization of eight genes is similar in that each comprises 14-15 exons. The choline transporter (CHT) is encoded in eight exons and the Na(+)-dependent myo inositol transporter (SMIT) in one exon. Mutations in three genes produce genetic diseases (glucose-galactose malabsorption, renal glycosuria and hypothyroidism). Members of this family are multifunctional membrane proteins in that they also behave as uniporters, urea and water channels, and urea and water cotransporters. Consequently it is a challenge to determine the role(s) of these genes in human physiology and pathology. PMID- 12748862 TI - Actin filaments regulate the stretch sensitivity of large-conductance, Ca2+ activated K+ channels in coronary artery smooth muscle cells. AB - Using the inside-out patch-clamp technique, large-conductance Ca2+ -activated K+ channel (BK(Ca)) currents were recorded from coronary artery smooth muscle cells. Cytochalasin D, an actin filament disrupter, increased channel activity ( NP(o), where N is the number of channels and P(o) the open probability), and this increase was reversed by phalloidin, an actin filament stabilizer. NP(o) was also increased by colchicine, a microtubule disrupter, and decreased by taxol, a microtubule stabilizer. With the stepwise increase of negative pressure in the patch pipettes, the activity of BK(Ca) gradually increased: the maximum effect (527% increase in NP(o)) was achieved at -40 cmH(2)O and the half-maximum effect at -25 cmH(2)O. The increase in NP(o) in response to negative pressure was abolished by phalloidin but not by taxol. These results imply that both actin filaments and microtubules inhibit the opening of BK(Ca) in coronary artery smooth muscle cells, but that only actin filaments are involved in the stretch sensitivity of BK(Ca). PMID- 12748861 TI - The SLC39 family of metal ion transporters. AB - SLC39 proteins are members of the broader ZIP family of metal ion transporters found in organisms at all phylogenetic levels. Most ZIP transporters have eight predicted transmembrane domains and a similar predicted topology. Their biochemical mechanism(s) of substrate transport are not yet known. Where characterized, these proteins have been found to transport metal ions from the cell exterior or lumen of intracellular organelles into the cytoplasm. Furthermore, members of the ZIP family have been implicated in the transport of zinc, iron, and/or manganese indicating that these proteins have diverse functions. There are 14 SLC39-related proteins encoded by the human genome. Studies of SLC39A1, SLC39A2, and SLC39A4, encoding the proteins hZip1, hZip2, and hZip4, have indicated roles in zinc uptake across the plasma membrane of various cell types. Genetic studies have specifically implicated SLC39A4 in the uptake of dietary zinc into intestinal enterocytes. Mutations in SLC39A4 have been identified in patients with acrodermatitis enteropathica, a genetic disease of zinc deficiency. PMID- 12748864 TI - Overall and differentiated ratings of perceived exertion at the respiratory compensation threshold: effects of gender and mode. AB - Research on gender differences in ratings of perceived exertion (RPE) has been equivocal with few studies comparing exercise modes and differentiated RPE. The current study examined gender differences in overall and differentiated RPE at the respiratory compensation threshold (RCT) during cycling and treadmill exercise. Each minute during a maximal treadmill and maximal cycling test, men (n=18) and women (n=16) estimated RPE corresponding to overall (RPE-O), legs (RPE L), and breathing/chest (RPE-C) exertion. A 2 (gender) x 2 (mode) x 3 (RPE-O, RPE L, RPE-C) repeated measures MANOVA revealed no significant mode x gender or RPE x gender interactions. The exercise mode x RPE interaction approached significance (P=0.055) when cycling [mean (SD) 14.8 (2.9)] and treadmill exercise [12.8 (2.9)] were compared. No main effects for gender [men: 13.7 (2.6), women: 13.4 (2.6)] were detected. Main effects for mode showed RPE to be significantly greater during cycling [14.4 (2.8)] versus treadmill exercise [12.7 (2.9)]. Main effects for differentiated RPE showed RPE-L [13.8 (2.6)] to be significantly greater than RPE-O [13.5 (2.6)] and RPE-C [13.3 (2.6)]. Results suggest that overall and differentiated RPE at the RCT are not significantly different between genders during cycling or treadmill exercise. While RPE-L was statistically greater than RPE-O and RPE-C, the magnitude of the differences makes this result of little practical significance. The marginal interaction suggests greater RPE-L values might be expected at the RCT during cycling versus treadmill exercise. However, results suggest that minimal RPE differences exist between men and women during cycling and treadmill exercise. PMID- 12748863 TI - A rapid enzymatic method for the isolation of defined kidney tubule fragments from mouse. AB - The increasing number of available genetically manipulated mice makes it necessary to develop tools and techniques for examining the phenotypes of these animals. We have developed a straightforward and rapid method for the isolation of large quantities of single tubule fragments from the mouse kidney. Immunohistochemistry, electron microscopy, and fluorescence microscopy were used to evaluate the viability, functional characteristics, and morphology of proximal tubules (PT), and collecting ducts from cortex (CCD) and inner stripe of the outer medulla (ISOMCD). Tubules were isolated using a modified collagenase digestion technique, and selected under light microscopy for experimentation. Electron microscopy and trypan blue exclusion showed that a large portion of unselected proximal tubules were damaged by the digestion procedure. The selected tubules, however, all excluded trypan blue, indicating that the plasma membrane had remained intact. Immunocytochemistry on isolated CCD showed normal distribution of H(+)-ATPase, pendrin, and anion exchanger-1 (AE-1) staining. The pH-sensitive dye 2',7'-bis(2-carboxylethyl)-5(6)-carboxyfluorescein (BCECF) was used to measure Na(+)-dependent and -independent intracellular pH (pH(i)) recovery rates in PT, and in single intercalated cells of CCD and ISOMCD fragments. Na(+)-dependent pH(i)-recovery was 0.144+/-0.008 (PT), 0.182+/-0.013 (CCD), and 0.112+/-0.010 pH units/min. (ISOMCD). Na(+)-independent pH(i) recovery was found in all three segments (PT: 0.021+/-0.002, CCD: 0.037+/-0.002, ISOMCD: 0.033+/-0.002 pH units/min) and was sensitive to concanamycin. In summary, we have developed a new technique for rapid and straightforward preparation of large quantities of defined tubule fragments from mouse kidney. Using this technique, the first measurements of plasma membrane vacuolar H(+)-ATPase activities in mouse PT and collecting duct were made. This technique will facilitate further characterization of kidney function in normal and genetically manipulated animals. PMID- 12748865 TI - Fatal truck-bicycle accident involving dragging for 45 km. AB - Vehicle-bicycle accidents with subsequent dragging of the rider over long distances are extremely rare. The case reported here is that of a 16-year-old mentally retarded bike rider who was run over by a truck whose driver failed to notice the accident. The legs of the victim became trapped by the rear axle of the trailer and the body was dragged over 45 km before being discovered under the parked truck. The autopsy revealed that the boy had died from the initial impact and not from the dragging injuries which had caused extensive mutilation. The reports of the technical expert and the forensic pathologist led the prosecutor to drop the case against the truck driver for manslaughter. PMID- 12748866 TI - Validation of the X-chromosomal STR DXS6809. AB - This paper presents sequence and population genetic data of the microsatellite marker DXS6809 (GDB 365492) obtained from a German population sample ( n=725 chromosomes). DXS6809 is a highly polymorphic X-linked tetranucleotide polymorphism presenting 12 alleles in our population. Sequencing of 77 PCR products covering 12 alleles (by length), characterised DXS6809 as a marker with a complex repeat sequence structure. A polymorphism information content (PIC) of 0.825 and a mean exclusion chance (MEC) of 0.815 were obtained. A deviation from the Hardy-Weinberg equilibrium (HWE) could not be detected and male and female samples exhibited a similar allele distribution. Kinship testing revealed a typical X-linked inheritance and 2 mutations were found in 394 meioses. DXS6809 is located 90.18 Mb, i.e. 102.3 cM, from the Xp-telomere (Xp-tel), corresponding to Xq21.33. The presented data qualify DXS6809 as a useful supplement to the known forensic ChrX marker panel. PMID- 12748868 TI - Women's attitudes towards psychosocial support in labour in United Arab Emirates. AB - OBJECTIVE: The objective of this study was to determine women's attitudes and preferences regarding psychosocial support during childbirth in United Arab Emirates (UAE). METHODS: A consecutive sample of 400 mothers with singleton normal pregnancies delivered vaginally during 2 months was interviewed postpartum about their experience and satisfaction with supportive care during labour using structured questionnaire. RESULTS: Birth attendant continuously accompanied 237 (59.3%) participants including mother (59.5%), sister (31.2%), friend (7.2%), other family member (1.3%) or husband (0.8%). Preference in the no-companion group ( n=163, 40.7%) was health professional (56.4%), mother (25.8%), sister (16.6%) or husband (1.2%). Labour was significantly shorter ( P<0.0001) with less need for analgesia ( P<0.0001), oxytocin augmentation ( P<0.0001) and neonatal intensive care ( P=0.03) in the companion group. Rates of instrumental delivery, episiotomy and perineal tear were similar in both groups. Three hundred and fifty (87.5%) subjects felt that psychosocial support during childbirth is essential and best provided by non-professional attendant (companion group) or midwife/obstetrician (no-companion group). 59.3% and 19.7% of mothers, respectively, reported less satisfaction and negative feelings about their perinatal experience that was more frequent in the no-companion group ( P=0.001, P<0.0001; respectively). CONCLUSION: Perceptions, experiences and outcomes of companion support during childbirth in UAE, although relatively less available, are therefore not different from those described elsewhere. PMID- 12748869 TI - Primary hydatid disease of the quadriceps muscle: a rare localization. AB - BACKGROUND: Hydatidosis is a protozoal infestation which generally involves the liver and the lungs. Primary skeletal muscle hydatidosis without involving the thoracic and abdominal organs is extremely rare. METHODS: A 48-year-old farmer presented with a mass in the quadriceps muscle that had persisted for 10 years. Clinical and radiological investigations revealed a primary hydatid cyst of the vastus medialis of the quadriceps muscle. We did not find any visceral organ involvement. Wide excision was performed without destroying the cyst wall. Then 400 mg albendazole was given daily for 3 months postoperatively. RESULTS: At the 7th postoperative month, the patient was symptom-free, and the laboratory test results were in the normal ranges. CONCLUSION: Primary muscular hydatidosis is a rare disease and should be kept in mind in the diagnostic work-up of a cystic mass of a skeletal muscle. PMID- 12748867 TI - Head and neck intramuscular haemangioma: report of two cases with unusual localization. AB - Head and neck intramuscular haemangioma (IMH) is a benign, uncommon neoplasm arising most frequently in the masseter and trapezius muscles. It usually appears as an enlarging mass without specific clinical features and is therefore frequently misdiagnosed. We report two cases of IMH at unusual localizations: one involving the pterygoid muscles in a 30-year-old female and another one originating from the thyrohyoid muscles in a 38-year-old male. Preoperative evaluation included routine ENT examination, ultrasound (US) and CT scan. The preoperative clinical diagnoses were lymphangioma of the parapharyngeal space in the first case and atypical thyroid duct cyst in the second. Both patients were treated with open-neck approaches. The histopathologic diagnosis was capillary type IMH in the first patient and cavernous-type IMH in the second. Postoperative courses were uneventful, and the patients were discharged within a few days. Clinical and US follow-ups were negative at 50 and 20 months, respectively. Preoperative diagnosis of head and neck IMH is rendered difficult by its rarity and non-specific clinical findings. Ample surgical excision is required for an adequate cure due to the frequent microscopically infiltrative pattern of diffusion of such a tumor into skeletal muscles, and long-term follow-up is mandatory to promptly treat any recurrence. PMID- 12748870 TI - The influence of walking speed on kinetic and kinematic parameters in patients with osteoarthritis of the hip using a force-instrumented treadmill and standardised gait speeds. AB - BACKGROUND: It is difficult to identify objective parameters for assessing the joint function when dealing with the evaluation of orthopaedic procedures, especially endoprosthetic hip replacement. Clinical gait analysis enables parameters of force and movement to be quantified. However, the influence of gait speed on these parameters has hardly been taken into consideration so far. The objective of the present study was therefore to investigate the effect of gait speed on gait parameters and to simplify the clinical conditions in patients with osteoarthritis of the hip by determining a standardised gait speed. METHODS: A total of 28 patients with severe unilateral osteoarthritis of the hip were investigated at different gait speeds. The gait analysis equipment used consisted of an infinitely adjustable treadmill with force plates and an infrared video system. A special control mechanism permitted adjustment of the treadmill speed to a patient's self-determined pace. RESULTS: The mean gait speed of all patients with osteoarthritis of the hip was set at 2.20 km/h (0.61 m/s). Eight of the 10 gait parameters assessed increased significantly with changing gait speed. Pathological changes in gait patterns were found at the three gait speeds investigated, with the changes more accentuated at higher speeds. CONCLUSIONS: Dependence of gait parameters on gait speed could be concluded for a group of patients and for control subjects. Use of a force-instrumented treadmill is necessary for the setting of a standard gait speed, which should be set as high as achievable by patients without inducing pain and problems of coordination or balance. With the usage of standardised speeds, clinical gait analysis becomes easier to perform. Furthermore, one can assess the expected biomechanical advantages of newer prostheses, thus providing the surgeon with a basis for further decisions. PMID- 12748872 TI - DNA microarrays in clinical practice: past, present, and future. AB - Gene expression is a central concept in molecular biology and forms part of our knowledge of the role of genes in human diseases. Genome-wide monitoring of gene expression using DNA microarrays (allowing the simultaneous assessment of the transcription of tens of thousands of genes, and of their relative expression between normal cells and pathological cells) represents one of the latest breakthroughs in experimental molecular biology and provides unprecedented opportunity to explore the biological processes underlying human diseases by providing a comprehensive survey of a cell's transcriptional landscape. This revolutionary technology ultimately leads to the discovery of new biomarkers for disease diagnosis and prognosis prediction, and of new therapeutic tools. This paper provides an overview of microarray technology and describes some of its recent applications in medicine. PMID- 12748871 TI - Immunohistochemical and ultrastructural study of chordoid glioma of the third ventricle: its tanycytic differentiation. AB - A chordoid glioma in the third ventricle was studied immunohistochemically and ultrastructurally. In this report, special attention is paid to the histogenesis in relation to the pathological appearance and unique anatomic location of this tumor. Light microscopic and immunohistochemical findings were similar to those reported previously. Ultrastructurally, microvilli were frequently seen, but three types of abnormal cilia were rarely observed. Basement membrane around the tumor cells and microvessels was extensive. Poorly to moderately developed intermediate (adherent) junctions were frequently seen. Resemblance of these ultrastructural features of the tumor to embryonic tanycytes suggests the tanycytic differentiation of chordoid glioma. Neuroradiologically, all of the previously reported cases of chordoid gliomas seem to arise in the anterior part of the third ventricular floor. This region includes the lamina terminalis, infundibular recess and median eminence, which corresponds to a tanycyte-rich area. These findings suggest a tanycytic origin of chordoid glioma. PMID- 12748873 TI - Gene therapy for hematological malignancies. AB - Since cancer is the result of genetic mutations, it should be well suited for correction through gene therapy. Hematological malignancies in which human gene transfer has been performed are leukemias, lymphomas, graft-versus host disease after allogeneic bone marrow transplantation in leukemia, and multiple myeloma. Gene therapy may be used to induce or enhance an antitumor immunological reaction, to correct a genetic defect in the tumor cells, to render the malignant disease more susceptible to conventional therapies, to make the normal host cells more resistant to conventional therapies, or to track cells used for therapy. Gene therapy will probably be most valuable for the eradication of minimal residual disease after the use of conventional therapies. PMID- 12748874 TI - Human peritoneal macrophages in culture: a model for studying inflammatory disorders in vitro. AB - In this study we characterized a model of human peritoneal macrophages maintained in culture for up to 48 h that can be used to study different functions of this cell population in vitro. The cells remained viable and functionally active over time, with well-preserved phagocytic properties. They expressed a macrophage marker, CD14. Once in culture, human peritoneal macrophages secreted C1q and nitric oxide in a pattern described in murine, guinea pig, and rat peritoneal macrophages. The described model can be used to study physiology and pathophysiology of peritoneal macrophages in vitro, offering all the advantages of the use of a human cell population. PMID- 12748875 TI - Effect of macrophage colony stimulating factor overexpression on oxidative injury/resistance of RAW264.7 cells. AB - Oxidative injury to monocytes/macrophages is considered one of the key factors in atherogenesis. Macrophage colony stimulating factor (M-CSF) also plays an important role in the stages of atherosclerosis. Some researchers showed that M CSF accelerated pathological changes in the early stages of atherosclerosis. However, other reports suggested that exogenous M-CSF could prevent the progress of atherosclerosis. To further investigate the role of M-CSF in atherogenesis and to elucidate the effect of M-CSF on the oxidative injury to monocytes/macrophages, RAW264.7 cell lines overexpressing M-CSF were established by applying the lipofectin transfection method. The oxidative injurious effect of tert-butylhydroperoxide on the established cell lines was investigated. Two M-CSF transfected RAW264.7 cell lines secreted large amounts of M-CSF. Compared with the non-transfected RAW264.7 cells, M-CSF-overexpressing RAW264.7 cells were more vulnerable to oxidative injury. We conclude that M-CSF could aggravate the oxidative injury due to macrophages in some situations. PMID- 12748876 TI - The clinical role of interleukin-6 and interleukin-6 soluble receptor in human follicular fluids. AB - In order to investigate the role of interleukin-6 (IL-6) and interleukin-6 soluble receptor (sR) in human ovulation, we evaluated the concentrations in human follicular fluid and analyzed the correlation of IL-6 and IL-6 sR with oocyte maturation. The oocytes were obtained from the follicular fluid of 45 women undergoing in vitro fertilization and embryo transfer. The concentrations of IL-6 and IL-6 sR in follicular fluid were measured by ELISA. In addition, granulosa cells obtained from the follicular fluid were cultured and treated with forskolin and 12- o-tetradecanoylphorbol 13-acetate for 24-48 h. The concentration of IL-6 was significantly higher in the follicular fluid than in the serum (P<0.01). In contrast, the concentration of IL-6 sR was significantly lower in the follicular fluid than in the serum (P<0.001). The concentrations of IL-6 and IL-6 sR were significantly higher in the follicular fluid containing mature oocytes than in fluid containing immature oocytes (P<0.05). The production of IL-6 was markedly increased over the basal level after 24 h of treatment with forskolin (P<0.001) and 48 h of treatment (P<0.01) with cultured granulosa cells. Our data suggest that IL-6 and IL-6 sR may play an important role in follicular growth and development in human preovulatory processes. It is possible that IL-6 in particular may be regulated by cAMP. IL-6 and IL-6 sR might also be valuable biochemical markers in the evaluation of oocyte maturation. PMID- 12748877 TI - Anticardiolipin, glutamic acid decarboxylase, and antinuclear antibodies in epileptic patients. AB - To explore the hypothesis that raised anticardiolipin antibodies, glutamic acid decarboxylase, and antinuclear antibodies may be associated with epilepsy and/or pharmacoresistance, we studied titers in 74 epileptic patients and 50 controls. Epileptic patients were divided into two groups according to their response to anticonvulsant therapy. Group I included 52 children (30 females and 22 males with a mean age+/-SD of 7.0+/-2.4 years) suffering from different types of epilepsy who were treated with various anticonvulsants. Group II included 22 children (10 females and 12 males with a mean age of 6.2+/-3.6 years) suffering from therapy resistant epilepsy. We found that the prevalence of anticardiolipin antibodies was significantly higher in epileptic patients than in controls, while there was no significant difference between patients who were seizure free and those with uncontrolled epilepsy. No significant difference was found in glutamic acid decarboxylase antibodies between epileptic children and controls, and between patients who were seizure free and those with uncontrolled epilepsy. A significant difference in the incidence of antinuclear antibodies was found between epileptic children and controls, while no difference was found between well-controlled and drug-resistant epilepsy. In conclusion, the prevalence of anticardiolipin and antinuclear antibodies was higher in patients with epilepsy than in controls. There was no significant difference in serum glutamic acid decarboxylase antibodies between epileptic children and controls, and between patients who were seizure free and those with uncontrolled epilepsy. PMID- 12748878 TI - T cell activation and enhanced apoptosis in non-ST elevation myocardial infarction. AB - Recent studies have shown that inflammation plays a major role in coronary plaque destabilization and in the induction of thrombosis in acute coronary syndromes. The aim of this study was to evaluate circulating lymphocyte activation and apoptosis in patients with non-ST elevation myocardial infarction (NSTEMI) in comparison with subjects with stable angina and with age-matched healthy controls. We considered T cell subpopulations, T cell surface HLA-DR and CD69 expression (evaluated by flow cytometry), lymphomonocyte spontaneous apoptosis (evaluated by ELISA), and IL2 production (evaluated by ELISA) in peripheral blood within 6 hours of onset of NSTEMI. We also investigated Fas expression on T cells (evaluated by flow cytometry) and FasL mRNA (evaluated by RT-PCR), as well as Fas functionality. In NSTEMI patients we found a significant increase of HLADR+ CD3+ and CD69+CD4+ cells. Spontaneous apoptosis was significantly increased in NSTEMI patients in comparison with the two control groups and was associated with an increased expression of Fas, an increased susceptibility to Fas agonist (CH11), and a normal production of IL2 in cell cultures. These data suggest that the enhanced apoptosis is due to a mechanism of "active" antigen-driven death, induced by the expression of death cytokines and not by the failure of cell growth factors. We conclude that peripheral lymphocytes are activated in NSTEMI and undergo an enhanced programmed cell death due to activation mechanisms. It is likely that lymphocyte activation occurs before the onset of acute ischemia and contributes to the plaque rupture and to the myocardial ischemic insult. PMID- 12748879 TI - Different effect induced by treatment with several statins on monocyte tissue factor expression in hypercholesterolemic subjects. AB - Platelets and monocytes are involved in atherothrombosis via tissue factor expression. Moreover, they are activated in hypercholesterolemia, a classic risk factor for atherothrombosis. Cholesterol-lowering drugs (statins) reduce cardiovascular risk either by decreasing cholesterol or non-lipidic actions, such as platelet and monocyte activity. The aim of our study was to evaluate the effect of several statins on platelet and monocyte activity in hypercholesterolemic subjects. Platelet activity (P-selectin, cytofluorimetric detection), tissue factor levels (ELISA) and activity (detected in whole blood and cellular preparations by a specific clotting assay) were measured in hypercholesterolemic subjects (41 males, 23 females, aged 34-65 years, total cholesterol 6.86+/-0.60 mmol/l) treated with atorvastatin 10 mg, simvastatin 20 mg, fluvastatin 40 mg, or pravastatin 40 mg for 6 weeks. P-selectin and tissue factor expression in whole blood and isolated cells were increased in hypercholesterolemic subjects with respect to controls (all P<0.001). Simvastatin, atorvastatin, and fluvastatin reduced monocyte procoagulant activity in whole blood and P-selectin (P<0.01). Tissue factor antigen and activity in isolated cells were further reduced (all P<0.05) independently of cholesterol lowering. Pravastatin decreased tissue factor expression in whole blood in direct relationship to reduction of P-sel and cholesterol (P<0.05). Our data show a different impact of several statins on monocyte tissue factor expression in whole blood, suggesting a possible role of decreased platelet activity and a direct action on monocytes. In contrast, pravastatin decreased monocyte procoagulant activity with relation to cholesteroldependent modifications of platelet function. PMID- 12748880 TI - Culture collections over the world. AB - Culture collections have the crucial role of providing the authenticated biological material upon which high quality research is based. Importantly, they serve as repositories for strains as part of patent deposits, providers of safe and confidential services to store key organisms for research and industry, and sources of organisms cited in scientific papers that can be used in the confirmation of results and for further study. The demands upon culture collections change as new technologies and uses of organisms are discovered. Many are becoming Biological Resource Centres, as defined by the OECD Biological Resource Centre (BRC) Initiative, in that they operate according to international quality criteria, carry out essential research, enhance the value and applications of strains and provide a vital information resource. In a changing international scientific environment, many collections are under threat of extinction because of inadequate funding, changing government support strategies and the cost of new technologies. We are also suffering a decline in the number of biosystematists, who are needed to form a sound base for molecular technologies and to aid in identifying, and characterizing microbial diversity. In this environment, collections must work together to make the best use of new technologies and to contribute to the description of the 1.4 million fungi yet to be discovered. At the current rate, this will take 700 years. New technologies and novel ways of funding this task must be engaged and, above all, scientists must collaborate. Common policies are necessary to address the regulatory demands on collections, to control access to dangerous organisms, and, in particular, to enforce the Convention on Biological Diversity. Countries that hold the majority of biodiversity require support in building the facilities required to explore their hidden resource. The World Federation for Culture Collections (WFCC) and, in Europe, the European Culture Collection Organisation (ECCO) have a key role to play. The world must benefit from its microbial diversity, which is crucial to solving increasing problems in food provision, public health and poverty alleviation. PMID- 12748882 TI - Quality control in neurosurgery training. PMID- 12748881 TI - Anion concentration modulates the conformation and stability of the molten globule of cytochrome c. AB - Anions induce collapse of acid-denatured cytochrome c into a compact state, the A state, showing molten globule character. Since structural information on partially folded forms of proteins is important for a deeper understanding of folding mechanisms and of the factors affecting protein stabilization, in this paper we have investigated in detail the effects of anions on the tertiary conformation of the A-state. We have found that the salt-induced collapse of acid denatured cytochrome c leads to a number of equilibria between high-spin and low spin heme states and between two types of low-spin states. The two latter states are characterized by conformations leading to a native-like Met-Fe-His axial coordination and a bis-His configuration. The equilibrium between these two A states is dependent on the concentration and/or size of the anions (i.e. the bigger the anion, the greater its effect). Further, on the basis of fast kinetic data, a kinetic model of the folding process from the acid-unfolded protein to the A-state (at low and high anion concentration) is described. PMID- 12748883 TI - CSF shunt infections in children: experiences from a population-based study. AB - The objective was to identify risk factors for shunt infections, and establish the rate of infection for shunt procedures carried out under standardized conditions in a well-defined population. All (407) paediatric shunt operations (primary and revisions) performed within a total population of 630000 inhabitants between January 1, 1986 and December 31, 1996, were analysed retrospectively. 11 shunt infections were diagnosed in 10 patients, giving an overall infection rate of 2.7% per procedure and 6.2% per patient. Infections were significantly correlated with age, type of operation, and a etiology of hydrocephalus. Thus, infections occurred more frequently during the first 6 months of life, more often following primary shunt insertions compared with revisions, and children with myelomeningocele had a higher infection risk than children with hydrocephalus due to other causes. There was a highly significant male preponderance in the patient material. CONCLUSION: The overall infection rate was relatively low. The risk factors for shunt infections appear to relate to epidemiological characteristics rather than to surgical factors. PMID- 12748884 TI - Direct costs of microsurgical management of radiosurgically amenable intracranial pathology in Germany: an analysis of meningiomas, acoustic neuromas, metastases and arteriovenous malformations of less than 3 cm in diameter. AB - BACKGROUND: The purpose of the analysis was to appreciate and compare the effective direct costs of microsurgical treatment of intracranial pathology potentially amenable to radiosurgery as they arose in 1998-99. METHOD: Treatment costs of 127 microsurgically treated patients harbouring an arteriovenous malformation (AVM), acoustic nEuroma, meningioma or brain metastasis potentially amenable to radiosurgery were reviewed. Costs for the surgical procedure, ICU care, medical and nursing care on the ward, interclinical bills (ICB) for services provided by other departments and the overhead for basic hotel service were added. For comparison Gamma Knife costs were calculated by dividing the global operating cost of the Gamma Knife centre by the number of patients treated in 1999. FINDINGS: Average hospitalisation time for the entire microsurgical patients was 15,4+/-8,6 days. The patients spent an average of 1,2+/-2,8 days on ICU. Average operating time for all patients, including preparation, was 393+/ 118 minutes. Average costs for the microsurgical therapy were Euro10.814+/-6.108. These consisted of Euro1417+/-426 for the surgical procedure, Euro1.188+/-2.658 for ICU care, Euro2.333+/-1.582 for medical and nursing care on the ward, Euro1.671+/-1.433 for interclinical bills and Euro 4.204+/-2.338 for basic hotel service (overhead, Euro273/day). 70% of the microsurgically treated patients needed ancillary inpatient rehabilitation or radiotherapy resulting in an average additional cost for all patients of Euro2.744. Furthermore 20% of the microsurgically treated patients required an unplanned readmission after discharge, resulting in an average additional costs for all patients of Euro1.684. Average overall costs per patient including ancillary therapy and unplanned readmissions amounted to Euro15.242. For comparison, Gamma Knife treatment costs per patient amounted to Euro7.920 in 1999. INTERPRETATION: The current analysis showed that for established radiosurgical indications the primary costs of microsurgery exceeded the costs of radiosurgery. Differences with regard to additional expenses as a consequence of disability were not addressed in this study. Microsurgical management as well as Gamma Knife radiosurgery have potential for economic improvement. PMID- 12748886 TI - Craniocervical junction instability: instrumentation and fusion with titanium rods and sublaminar wires. Effectiveness and failures in personal experience. AB - BACKGROUND: The aim of the study was to evaluate the effectiveness, pitfalls and failures of instrumentation and fusion with titanium wires and rods in 12 h patients with craniovertebral junction instability. METHODS: Among nine adult patients (mean age 48.11 years) with craniovertebral junction instability, four had basilar impression, three metastatic disease, one rheumatoid arthritis and one Down's syndrome. Three children (mean age 7.33 years) with genetic (Down's syndrome, 2 cases) and metabolic (mucopolisaccarydoses type IV, i.e. Morquio Syndrome, 1 case) disease were studied as well. Each patient underwent preoperative radiological evaluation by means of X-Ray, CT scan and MRI of the craniocervical region. Occipitocervical instrumentation with a titanium U-shaped wired rod was performed in each patient. Autologous bone fusion was performed in all but the two cancer patients, in whom polymethylmetacrylate was used. Postoperatively, all the patients used an external orthosis for 3-6 months. Post operative X-Ray, CT and MRI were performed on each patient. The Frankel clinical scale was used to asses the outcome at follow-up which ranged from 1 to 10 years. At maximum follow up, there was either clinical improvement or stabilization recorded in all but one patient. This patient with basilar impression transiently worsened from grade D to C and a spinal cord lesion was already evident before the operation on MRI examination. INTERPRETATION: The effectiveness of surgical management of craniovertebral junction instability by instrumentation and fusion was demonstratedly in our experience. Nevertheless, the choice of the surgical technique should be made with caution when a spinal cord lesion is revealed by preoperative neuroimaging studies. PMID- 12748885 TI - Nitric oxide metabolites in cisternal CSF correlate with cerebral vasospasm in patients with a subarachnoid haemorrhage. AB - BACKGROUND: The pathogenesis of cerebral vasospasm is likely to be multifactorial. Exposure of the adventitia of large cerebral arteries to blood breakdown products initiates a cascade of changes in both morphology and vasomotor regulation of the exposed vessels. The role of nitric oxide (NO) in development of cerebral vasospasm process is controversial. Basal cerebral vascular tone requires the continuous release of NO, nevertheless NO is involved in free radical mediated injury of endothelial cell membrane. Concentrations of nitrate/nitrite (stabile endproducts of NO metabolism) were studied in cisternal cerebrospinal fluid (cCSF) in patients suffering from aneurysmal subarachnoid haemorrhage (SAH). METHOD: 21 patients suffering from aneurysmal SAH were investigated. Treatment included aneurysm clipping, cisternal drainage of CSF and intravenous nimodipine in all patients as well as tripple H therapy when indicated. TCDS was performed on a daily basis. A mean flow velocity of more than 150 cm/sec and the development a delayed neurological deficit was defined as vasospasm. CSF samples were collected on the day of surgery and for the 7 days following. NO-M (nitrite and nitrate) were measured using a commercially available test kit. FINDINGS: 5 of 21 patients developed clinically symptomatic vasospasm. There was a significant difference in NO levels between the groups. Patients with cerebral vasospasm showed significantly higher levels of NO-M in CSF than patients with a uncomplicated follow-up between day 2 and 8. INTERPRETATION: Our preliminary results indicate that SAH leads to an increase in NO-M in CSF. This increase of NO-M significantly correlates with the flow velocities in TCDS measurement suggesting that NO plays an important role in the pathogenesis of cerebral vasospasm. PMID- 12748887 TI - Large sphenocavernous meningiomas: is there still a role for the intradural approach via the pterional-transsylvian route? AB - BACKGROUND: Large-sized sphenocavernous meningiomas represent a surgical challenge. Although the role of skull base techniques with combined extra- and intradural steps has been recently emphasized, pure intradural resection tactics via the pterional route constitute the traditional microsurgical approach for resection of such tumours. METHOD: We report the application of the pterional transsylvian approach in 13 patients with sphenocavernous meningiomas. This series is unique because it includes only patients with tumours exceeding 5 cm in their greatest dimension. FINDINGS: A gross total resection was accomplished in 10 patients (77%). Eight patients had a good outcome, one had a persistent mild hemiparesis, and one died. No recurrences occurred in this group. Three patients (23%) had subtotal resections owing to invasion of the cavernous sinus in one instance and encasement of the middle cerebral artery in the others. Two had a good outcome and one died. In these patients minimal asymptomatic tumour progression was seen 3 and 6 years after surgery. The overall surgical outcome was good in 10 patients (77%), fair in one, and death in two. INTERPRETATION: In our experience, large sphenocavernous meningiomas may be operated on adopting pure intradural resection tactics via the pterional-transsylvian route with rates of gross total removal and surgical complications related to brain retraction or vascular manipulation comparable to those of extensive skull base approaches. The traditional intradural pterional transsylvian approach continues to have a place in the treatment of these lesions. PMID- 12748888 TI - Extended pterional orbital decompression in severe orbital cellulitis. AB - BACKGROUND: Bacterial orbital cellulitis is a relatively uncommon infective process, which can threaten the function of orbital structures. Apart from antibiotic therapy, sinus surgery with or without abscess drainage via an orbito otorhinolaryngological approach might be necessary. CASE DESCRIPTION: We present three cases of severe orbital cellulitis, leading to increasing loss of vision, proptosis, afferent pupillary disturbances and restriction of extra-ocular movements, despite antibiotic therapy. After extended pterional orbital decompression and reducing the orbital pressure by removal of the lateral and superolateral orbital walls, all patients showed distinct improvement of initial symptoms, without any complications related to the operation. INTERPRETATION: Extended pterional orbital decompression represents an effective treatment alternative and supplement in cases of a severe, threat to ocular function due to orbital cellulitis, where acute reduction of pressure on orbital, neural and vascular structures is intended. PMID- 12748889 TI - Simple invasive fixation device for fractionated stereotactic LINAC based radiotherapy. AB - BACKGROUND: The aim of this study was to develop a relocatable fixation device for linear accelerator (LINAC) based fractionated stereotactic radiotherapy. METHOD: The device consists of a CT- and MRI-compatible stereotactic frame, monocortical titan bone screws, four frame-posts with a lock for the fixation pins and a modified head clamp with additional arms that allows the exact and rigid placement of the frame in the desired final position prior to the final placement of the bone screws. By simply disconnecting the lock from the posts, the frame can be dismounted after treatment planning and after each treatment session. The accuracy of reposition was assessed prospectively, using phantom studies and also by comparison of isocenter movements during fractionated radiotherapy in 10 patients with an intracranial lesion. FINDINGS: No adverse events were seen after the surgical procedure and the screws were well tolerated throughout the course of treatment. The mean isocenter shifts observed during phantom reposition studies were x=0.05 mm, y=-0.32 mm, z=0.18 mm and the mean isocenter shifts during fractionated treatment were x=0.67 mm, y=0.65 mm, z=0.44 mm. INTERPRETATION: This new fixation device provides excellent accuracy of reposition during stereotactic radiotherapy. It appears superior to non-invasive, mask fixation techniques. Safety margins as small as 1-1.5 mm may therefore be sufficient for this method of stereotactic radiotherapy. PMID- 12748890 TI - The arcade of Struthers: an anatomical study. AB - BACKGROUND: The arcade of Struthers has been described as possible cause of compression of the ulnar nerve but several authors and our own experience have doubt cast on its existence. We, therefore, performed an anatomical study to determine whether or not the arcade of Struthers exists. METHODS: Ten arms from fresh frozen cadavers were dissected. Special attention was given to the fascial layers of the medial aspect of the upper arm and the medial intermuscular septum and their relation ships to the ulnar nerve. FINDINGS: An arcade of Struthers was not found in any specimen. INTERPRETATION: The arcade of Struthers does not exist and is not an anatomical structure, but a man-made construct. PMID- 12748891 TI - Do rapid systemic changes of brain temperature have an influence on the brain? AB - BACKGROUND: The purpose of the present study was to examine the influence of cooling and rewarming conditions using an accurate brain temperature control system. METHOD: The brain temperature of animals was measured with a thermometer while feedback regulation was achieved with a cold (4( degrees )C) and hot (50( degrees )C) water on-off flow system. Brain temperature was well controlled throughout the experiment by using both cold water and hot water simultaneously. Three groups were studied, as follows: 1) the standard group (cooled to 24( degrees )C for 1 hour, kept at 24( degrees )C for 2 hours and rewarmed to 37( degrees )C for 1 hour), 2) the rapid-cooling group (cooled to 24( degrees )C for 30 min, kept at 24( degrees )C for 2 h, and rewarmed to 37( degrees )C for 1 h), 3) the rapid-rewarming group (cooled to 24( degrees )C for 1 h, kept at 24( degrees )C for 2 h, and rewarmed to 37( degrees )C for 30 min) and the normal control group. FINDINGS: An increase of MAP-2 immunoreactivity of the CA1 neurons in the dorsal hippocampus was observed one week but not one month after hypothermia in the rapid-rewarming group. There was also a significant increase in the glutamate and lactate value at the end of rewarming compared with the baseline in the rapid-rewarming group (p<0.01). INTERPRETATION: Our results suggest that rapid rewarming after hypothermia triggered an uncoupling of cerebral circulation and metabolism, inducing an increase of extracellular glutamate and lactate, consequently reversible neuronal cell damage. PMID- 12748892 TI - Bilateral meningiomatous lesions of the spinal accessory nerves. AB - BACKGROUND: Meningiomas arising from cranial nerves with no dural attachment are exceedingly rare. The authors present a patient with bilateral meningiomatous lesions originating symmetrically from both spinal accessory nerves. CASE REPORT: A 61-year old woman presented with a one-year history of spinal ataxia and minimal left-sided motor impairment. Magnetic resonance imaging demonstrated two extrinsic lesions dorsolaterally of the medulla. Surgical exposure via a midline suboccipital approach with C1 laminectomy revealed the lesions arising from the spinal accessory nerves and in direct contact with the vertebral arteries. Histological investigation showed hypocellular fibrous lesions with proliferating meningothelial cells, psammoma bodies and immunoreactivity for vimentin, S-100 protein and epithelial membrane antigen. INTERPRETATION: To the authors' knowledge this is the first report of intradural tumours of the spinal accessory nerves not derived from Schwann cells and the first report of bilateral intracranial meningiomatous lesions without dural attachment. PMID- 12748893 TI - Idiopathic symptomatic epidural lipomatosis of the lumbar spine. AB - BACKGROUND: Symptomatic spinal epidural lipomatosis (SEL) of the lumbar spine is a rare disease, often associated with steroid overload. Idiopathic lipomatosis is even much less frequent. Signs and symptoms depend upon the level and degree of nerve root compression. Diagnosis is best based on MRI. Weight reduction can be curative, however after failure of medical treatment or in severe cases surgical decompression should be performed. METHOD: Four patients with severe symptoms of lumbar spinal epidural lipomatosis were treated by surgical decompression. Patient history and neurological examination are described, diagnostic imaging is demonstrated, surgical treatment and outcome are documented. Different surgical techniques including laminectomy, interlaminar fenestration and lateral recess decompression were applied and are discussed. FINDINGS: All four patients improved after surgery. No surgical complications were observed. Even though limited to four cases this is the second largest series of operated idiopathic spinal epidural lipomatosis. INTERPRETATION: Surgical decompression was effective in improving symptoms in severe lumbar idiopathic spinal epidural lipomatosis. PMID- 12748894 TI - Multifocal intradiploic cavernous hemangioma of the skull associated with nasal osteoma. AB - OBJECTIVE: To present a 42-year-old female patient with multifocal cavernous hemangioma of the skull associated with nasal osteoma. DESIGN: A case report. INTERVENTION METHODS: X-rays, computerized tomography, magnetic resonance imaging, and histopathology were used to achieve the diagnosis of this rare entity. The multiple cavernous hemangiomas were resected en-bloc and a curettage biopsy was obtained from the nasal osteoma. RESULTS: The patient healed well after the operation. No recurrences of the cavernous hemangiomas were observed after one-year follow-up. CONCLUSION: Multifocal cavernous hemangiomas are rare benign lesions of the calvarium, arising from the intrinsic vasculature of the bone. Although they are benign, radiological findings are not always characteristic and their multiple presentation may easily make surgeons consider the other malignancies of the skull in the differential diagnosis. Histopathologic confirmation of the tumor is the definitive method for diagnosis. The treatment of choice is early en bloc resection of the tumour where it is possible. PMID- 12748895 TI - Non-rigid immobilisation of odontoid fractures. AB - Despite various reports on the management of odontoid fractures, there is no consensus on the subject, and the appropriate treatment still remains controversial. While untreated fractures or fractures treated only with a cervical orthosis seem to have the highest rate of non-union, the need for rigid external stabilisation has never been substantiated. In a retrospective analysis we reviewed 26 patients with acute type II and III fractures of the odontoid, treated with a cervical orthosis only. Study inclusion was limited to fractures that had a fracture gap of less than 2 mm, an initial antero-posterior displacement of less than 5 mm and angulation of less than 11 degrees, less than 2 mm displacement on lateral flexion/extension views, and were without neurological deficits. These fractures were defined as stable. There were 19 (73.1%) type II and 7 (26.9%) type III fractures; in 10 (38.5%) of these fractures the odontoid was displaced and/or angulated. The overall complication rate was 11.4% ( n=3). One patient suffered from pulmonary embolism, in two patients (7.7%) with initially minimally displaced fractures, secondary internal stabilisation had to be performed because of persistent instability. In 20 (77%) of the remaining fractures healing was uneventful. In 4 nondisplaced fractures (15%) fibrous union was documented. Three of these patients were over 65 years old. The overall fusion rate was 73.7% for type II and 85.7% for type III fractures. At follow-up 39% of the patients were free of symptoms; however, the clinical outcome did not correlate with the radiological findings. According to our findings, stable type II and type III fractures of the odontoid can be successfully treated with non-rigid immobilisation, even if they are displaced. A thorough assessment of the stability of the odontoid with lateral flexion/extension views or dynamic fluoroscopy is recommended to evaluate the appropriate treatment. Non-rigid immobilisation may be an option in selected cases with stable injuries. PMID- 12748896 TI - Back pain remains a common symptom in old age. a population-based study of 4486 Danish twins aged 70-102. AB - Back pain (BP) has been rated among the most important factors affecting physical health status in old age. Yet there is an under-representation of the older population in the BP literature. We present extensive interview data from the Longitudinal Study of Aging Danish Twins, dealing with a population-based sample of Danish twins aged 70-102, and describing the 1-month prevalence of BP and the development of BP over time. The associations between BP and education, self rated health, other health problems, lifestyle factors, and physical and mental function were also investigated. Data were analysed in a cross-sectional analysis for all answers given at entry into the study and in a longitudinal analysis for participants in all four surveys. Associated factors were analysed for the cross sectional sample using univariate and multivariate analysis accounting for the non-independence of twins in complete pairs. The overall 1-month prevalence of BP was 25% and differed significantly between men and women. The variations in prevalence between the age groups and over time were negligible. The majority of participants in all four surveys had either not experienced BP during the previous month or had done so on one occasion only. Education was not associated with BP. Self-rated health was associated with BP in a significant "dose response" like pattern. BP was associated with bone and joint disorders, migraine headaches, lung disease, cardiovascular disorders and gastric ulcer, but not neurologic or endocrinologic diseases. BP sufferers had significantly lower scores on physical but not on mental functioning. We conclude that BP is a common symptom in old age; however, the prevalence does not change with increasing age. BP may be part of a more general syndrome of poor health among the old. PMID- 12748897 TI - Spinal fusion in patients with Duchenne's muscular dystrophy and a low forced vital capacity. AB - Traditionally, spinal fusion has been denied to patients with scoliosis secondary to Duchenne's muscular dystrophy (DMD) when their forced vital capacity (FVC) is less than 30-40% of predicted values (PFVC). The reasons for this decision are a theoretically increased risk of adverse events from a prolonged anaesthetic and extensive surgery. This paper presents a retrospective analysis of 30 patients with DMD scoliosis who underwent posterior spinal fusion at the Royal National Orthopaedic Hospital. Two subgroups of patients were compared: those with more than 30% PFVC (17 patients) and those with less than 30% PVFC (13 patients). One patient in each group required a temporary tracheotomy and there were nine complications in total. The post-operative stay for patients in each group was similar (24 days in the >30% group, 20 days in the <30% group) and the complication rate was comparable with other published series. We conclude that spinal fusion can be offered to patients with DMD even in the presence of a low FVC. PMID- 12748898 TI - [SARS--episode or catastrophe?]. PMID- 12748899 TI - [Clinical and angiographic one-year follow-up of vascular beta-brachytherapy for coronary lesions treated by a stent with a very high risk for restenosis]. AB - BACKGROUND: Vascular brachytherapy (VBT) has been proven to reduce restenosis rate and unwanted cardiac events in several randomized trials. Long-term data on populations at high risk for re-interventions are few. The aim of this study was to assess the acute and one-year outcome of beta-radiation in coronary in-stent restenoses with a high likelihood of recurrence. METHODS: In 79 patients, VBT using 90Yttrium/Strontium or 32Phosphorus, was performed. Clinical and angiographic follow-up was carried out after 6 months and 1 year. RESULTS: 44.4 % of patients had three-vessel coronary artery disease and a high prevalence of cardiovascular risk factors and comorbidity. Mean lesion length was 36.8+/-18.9 mm. VBT was successful in all patients. Fractionation of VBT was necessary in 2,5 %. Acute gain of luminal diameter was 2.15+/-0.89 mm. During the hospital stay one acute myocardial infarction (AMI) not associated with VBT occurred. After 6 months loss of luminal diameter measured 0.39+/-0.47 mm, equaling a restenosis rate (RR) of 16.8 % (1 year: 0.60+/-0.56 mm, RR 33.5 %). 18.9 % of patients required revascularization of the target lesion (1 year: 29.5 %). After 6 months, all patients survived, three had an AMI after discontinuation of clopidogrel, one of them was asymptomatic (1 year: 1 cardiac death, 2 symptomatic AMI). CONCLUSION: Beta-VBT in patients at a high risk for recurrence after angioplasty is feasible and safe. Though the clinical and angiographic results at 1 year showed some impairment as opposed to the 6-months-follow-up, they nevertheless are largely superior to those patients from historic controls not treated with VBT. PMID- 12748901 TI - [Persistent polyclonal B-cell lymphocytosis]. AB - HISTORY AND CLINICAL FINDINGS: A 54-year-old woman was referred for ambulant checkup after an episode of acute renal failure due to severe gastroenteritis and recurrent arthralgias. Physical examination was unremarkable except for the presence of palpable small cervical lymph nodes. INVESTIGATIONS: Serum IgM levels showed a polyclonal increase. All the other routinely examined parameters were within normal limits. Microscopical blood smear examination revealed binucleated lymphocytes. Immunophenotyping of peripheral blood showed a polyclonal B-cell lymphocytosis despite normal numbers of leukocytes and lymphocytes. PCR analysis identified cells with a t(14;18) translocation (bcl-2/IgH rearrangement). DIAGNOSIS: A routine medical checkup disclosed the diagnosis of persistent polyclonal B-cell lymphocytosis. This rare benign lymphoproliferative disorder is characterized by binucleated lymphocytes, polyclonal expansion of B-cells, and a polyclonal increase in serum IgM. The diagnosis was established despite the lack of leukocytosis or lymphocytosis in the peripheral blood. CONCLUSIONS: Because of its benign and indolent course without the need for chemotherapy, it is important to discriminate the disorder of persistent polyclonal B-cell lymphocytosis from other malignant lymphoproliferative diseases. PMID- 12748902 TI - [Current diagnosis of poisoning]. PMID- 12748900 TI - [Clinical presentation and management of the severe acute respiratory syndrome (SARS)]. AB - BACKGROUND AND OBJECTIVE: In February 2003, a newly emerged infectious disease was described, the etiology of which was initially unknown. It is referred to under the term SARS. In the beginning, it spread in some regions South-East Asia. Import infections appeared in many other parts of the world. Based on the first cases in Germany, this report illustrates the clinical appearance, the diagnostic results and the management of this new disease. PATIENTS AND METHODS: We analysed the data of two patients with SARS and one suspected patient. The results of radiological, laboratory, microbiological and physical examinations were abstracted and compared with the data obtained in other regions. RESULTS: Two of the three patients under our care developed SARS disease. This is characterised by fever of sudden onset lasting for more than 5 days, rapidly changing consolidations in chest x-ray not affected by antimicrobial therapy, leuko-, lympho- as well as thrombopenia with a compromised pulmonary function later in the course. Close contacts with SARS patients does not regularly result in full development of the disease. Secretion of a coronavirus could be detected in respiratory samples during the febrile phase and in feces for a longer time. It is still an open question whether bedrest and antibiotic prophylaxis by themselves or an additional administration of ribavirin and corticosteroids can improve the outcome. CONCLUSION: SARS is a new and highly contagious lung disease. It is crucial to be able to recognize the clinical appearance and the diagnostic features of this disease at an early stage, in order to prevent a further dissemination of the disease. PMID- 12748903 TI - [Obstructive sleep apnea, autonomic dysfunction and cardiovascular risk]. PMID- 12748904 TI - [Use of low-molecular-weight heparins in patients with prosthetic heart valves]. PMID- 12748905 TI - [Fatty liver and serum lactate elevation in an HIV patient--is the routine determination of thiamine and riboflavin during HAART useful?]. PMID- 12748906 TI - Two modes of germline instability at human minisatellite MS1 (locus D1S7): complex rearrangements and paradoxical hyperdeletion. AB - Minisatellite MS1 (locus D1S7) is one of the most unstable minisatellites identified in humans. It is unusual in having a short repeat unit of 9 bp and in showing somatic instability in colorectal carcinomas, suggesting that mitotic replication or repair errors may contribute to repeat-DNA mutation. We have therefore used single-molecule polymerase chain reaction to characterize mutation events in sperm and somatic DNA. As with other minisatellites, high levels of instability are seen only in the germline and generate two distinct classes of structural change. The first involves large and frequently complex rearrangements that most likely arise by recombinational processes, as is seen at other minisatellites. The second pathway generates primarily, if not exclusively, single-repeat changes restricted to sequence-homogeneous regions of alleles. Their frequency is dependent on the length of uninterrupted repeats, with evidence of a hyperinstability threshold similar in length to that observed at triplet-repeat loci showing expansions driven by dynamic mutation. In contrast to triplet loci, however, the single-repeat changes at MS1 exclusively involve repeat deletion, and can be so frequent--as many as 0.7-1.3 mutation events per sperm cell for the longest homogeneous arrays--that alleles harboring these long arrays must be extremely ephemeral in human populations. The apparently impossible existence of alleles with deletion-prone uninterrupted repeats therefore presents a paradox with no obvious explanation. PMID- 12748908 TI - [Psychosocial distress and symptoms of single mothers and their children in a large community sample]. AB - OBJECTIVES: As in other western countries the proportion of single parent families in Germany is growing. International studies showed lower socioeconomic status and elevated psychic distress in single mothers as well as increased problem behavior in their children. This study was performed to make up for the lack of epidemiologic investigations on this issue in Germany. METHODS: The family status of a complete cohort of school beginners in Duesseldorf was screened (recruitment rate 97.5%, N = 5,048). Single mothers and their children were compared to a control group of married mothers (N = 278) with respect to social variables and psychic distress (SCL-90-R, CBCL). RESULTS: 907 (18.0%) of all children lived in single parent families. Socioeconomic status (income, education, receiving welfare) of single mothers (total N = 891) who fulfilled the inclusion criteria (N = 531) was lower, while psychic distress was significantly elevated (GSI 0.45 vs. 0.28). Single mothers without additional personal support for their child, younger as well as poor single mothers showed higher values of GSI. Statistically significant increased behavior problems were found only in boys of single mothers. CONCLUSIONS: Single mothers are socially disadvantaged. They show an elevated mean psychic distress compared to married mothers. Psychosocial support for higher impaired subgroups of single mothers should be developed. PMID- 12748907 TI - Association and interaction of the IL4R, IL4, and IL13 loci with type 1 diabetes among Filipinos. AB - In the search for genes involved in type 1 diabetes (T1D), other than the well established risk alleles at the human leukocyte antigen loci, we have investigated the association and interaction of polymorphisms in genes involved in the IL4/IL13 pathway in a sample of 90 Filipino patients with T1D and 94 controls. Ten single-nucleotide polymorphisms (SNPs), including two promoter SNPs in the IL4R locus on chromosome 16p11, one promoter SNP in the IL4 locus on chromosome 5q31, and four SNPs--including two promoter SNPs--in the IL13 locus on chromosome 5q31 were examined for association, linkage disequilibrium, and interaction. We found that both individual SNPs (IL4R L389L; odds ratio [OR] 0.34; 95% confidence interval [CI] 0.17-0.67; P=.001) and specific haplotypes both in IL4R (OR 0.10; 95% CI 0-0.5; P=.001) and for the five linked IL4 and IL13 SNPs (OR 3.47; P=.004) were strongly associated with susceptibility to T1D. Since IL4 and IL13 both serve as ligands for a receptor composed, in part, of the IL4R alpha chain, we looked for potential epistasis between polymorphisms in the IL4R locus on chromosome 16p11 and the five SNPs in the IL4 and IL13 loci on chromosome 5q31 and found, through use of a logistic-regression model, significant gene-gene interactions (P=.045, corrected for multiple comparisons by permutation analysis). Our data suggest that the risk for T1D is determined, in part, by polymorphisms within the IL4R locus, including promoter and coding sequence variants, and by specific combinations of genotypes at the IL4R and the IL4 and IL13 loci. PMID- 12748909 TI - [Anxiety and depression in intake interviews of East and West German psychotherapy patients]. AB - OBJECTIVES: The present paper describes a study on the intake interview narratives of psychotherapy patients with assisted content analysis methods. METHODS: 47 psycho-dynamic diagnostic interviews of patients suffering from depression or anxiety disorders were analysed using a Gottschalk-Gleser technique with the Dresdner Angstworterbuch (DAW) as well as with the Affektives Diktionar Ulm (ADU). RESULTS: Comparisons between different subgroups of this sample were made: Anxiety vs. depression patients, East Germans (Magdeburg) vs. West Germans (Dusseldorf) and men vs. women. The comparisons showed that feelings of anxiety were important in all subgroups. Depressed patients not only spoke more often about depressed feelings in comparison with the other subgroups, but also seemed more able to express emotions of other kinds. Among the differences according to the diagnostic dichotomy there were also significant results which demonstrated the influence of cultural background and sex of the interviewees on the contents of the intake interviews. CONCLUSIONS: The results confirm our hypothesis that the psychotherapist-patient-interaction sequences, which are the basis of diagnostic categorization, consist of elements specific for the disorders under study as well as elements specific for sex and culture (east vs. west). Some content analysis markers of these elements are identified in our study. PMID- 12748910 TI - [Disturbed regulation of self-esteem in patients with overt versus covert self destructive behaviour]. AB - OBJECTIVES: According to psychoanalytic models self-harming patients are characterised by an unstable self-system and a disturbed regulation of self esteem. This is presumed to be denied or dissociated to a greater degree by those who harm themselves secretly (factitious patients) as compared to those who show open self-harm. It is hypothesised and empirically tested that self-destructive patients have more profound disorders of narcissistic self-regulation than patients without self-destruction, and that this should be more evident in patients with overt self-destructive behaviour. METHODS: The sample consists of 354 psychosomatic patients, 32 of whom demonstrated self-destructive behaviour (18 exclusively overt and 6 exclusively covert types of behaviour, according to Willenberg et al.). The narcissism inventory was applied. RESULTS: Self destructive patients showed higher levels on the "threatened self"-dimension than psychosomatic patients without self-harm. Overtly self-harming patients showed a higher degree of narcissistic self-regulation than covertly self-destructive patients. CONCLUSIONS: This supports theoretical assumptions of a disturbed regulation of self-esteem in self-destructive patients, especially in overtly self-harming patients. PMID- 12748911 TI - [Decision-making capacity and informed consent in living organ donation- difficult constellations in the psychosomatic and medico-ethical assessment of potential donors]. AB - Organ donation from relatives or other persons with a close relationship to the patient has become a fairly routine procedure in many countries. However, donors should be chosen carefully not only to avoid medical or psychosocial risks but also ethically questionable outcomes. Prerequisites for living organ donation are reported together with a short review of the known risks for donors. Under the central maxim of averting harm for the donor, six cases are described to illustrate typical difficulties in the psychosomatic-ethical consultation-liaison service, and our own procedures are discussed. Suggestions are made as to how to approach the information, evaluation and selection of potential donors. PMID- 12748912 TI - [Body image and mental structure--the ascertainment of body experience in psychodynamic diagnostics]. AB - The present paper introduces a new method for diagnosing the body image and reference to the integrational level of personality structure. First the body image is defined using five criteria: consciousness, flexibility, intersubjectivity, representational links to thought contents, and level of structural integration of the personality. Using axis four of the Operationalised Psychodynamic Diagnostics (OPD) as a starting point, dimensions are defined and described that allow assessment of the body image according to structural dimensions of the personality. The structural diagnostics of the body image can serve as a basis for a new rating instrument of body image integration. PMID- 12748915 TI - Case study: left eyelid swelling and apnea in a neonate. PMID- 12748916 TI - Homeless adolescents: common clinical concerns. AB - Homeless youth are at alarmingly high risk for a myriad of physical and psychological problems as a result of both the circumstances that prededed their homelessness, and as a direct consequence of life on the streets. Sexually transmitted infections (STIs), pregnancy, trauma, tuberculosis, uncontrolled asthma, and dermatologic infestations are a few of the health problems with which these youth commonly present. These somatic problems are compounded by high rates of drug and alcohol abuse as well as depression and suicide. Despite the obvious need for medical services, homeless youth often do not receive appropriate medical care due to numerous individual and systems barriers impeding health care access by this population. In addition to the barriers experienced by the adult homeless population, homeless adolescents confront further hurdles stemming from their age and developmental stage. Some of these impediments include a lack of knowledge of clinic sites, fear of not being taken seriously, concerns about confidentiality, and fears of police or social services involvement. Improved access to appropriate health care is necessary if we are to better support and care for this population of young people. To effectively manage and treat homeless youth, individual providers must be aware of the diagnoses associated with homelessness, as well as the community resources available to these youth. Finally, providers need to be the voices advocating for improved services for this disadvantaged and silent population. PMID- 12748918 TI - Ectoparasitic infestations in homeless children. AB - Most human ectoparasites live on the surface of their host and depend on that host to complete their life cycle. The most common ectoparasitic infestations of medical importance in humans include pediculosis, scabies, myiasis, and tungiasis. Different host factors are related, with increased risk of acquiring ectoparasitic infestation occurring among the homeless. Although these ectoparasitic infections can be found worldwide, their prevalence is affected significantly by environmental conditions in different geographical areas. This review focuses on the epidemiology, clinical presentation, diagnosis, and treatment of common ectoparasitic infestations among homeless children and their families. The most frequent bacterial infections associated with these infestations also are discussed. PMID- 12748917 TI - Human immunodeficiency syndrome and hepatitis B and C infections among homeless adolescents. AB - The lifestyle of homeless adolescents places them at high risk for contracting a variety of serious illnesses. The purpose of this cross-sectional study was to determine the predictors of human immunodeficiency virus (HIV) and hepatitis infections among homeless youth. Anonymous and confidential interviewer administered psychosocial surveys were conducted with 150 homeless youth (ages 14 to 23 years; 70% male). Venous blood samples were collected as well. Most respondents (95%) had engaged in sexual intercourse, with 13 years as the median age at first coitus. Approximately 36 percent indicated they had exchanged sex for food, shelter, or drugs. Although youth reported a high rate of consistent condom use, 16 percent of the sample tested positive for HIV, 17 percent tested positive for hepatitis B (HBV), and 12 percent tested positive for hepatitis C (HCV). Homeless youth should be considered a high-risk group for contracting HIV, HBV, and HCV infections. The positive rates for HIV, HBV, and HCV observed in this study are higher than those of many previous reports among street youth. Because HIV prevention and hepatitis B vaccination programs have been targeted at school-based youth, most homeless youth are missed by these efforts. Targeted programs are needed to reach this high-risk group. Suggested guidelines for clinicians involved in the provision of medical care to homeless children and adolescents are provided. PMID- 12748919 TI - Children orphaned by AIDS: a global perspective. AB - Internationally, the orphan crisis caused by the human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) pandemic remains a serious issue with long-term social consequences. At the end of 2001, an estimated 14 million children worldwide had lost their mother or both parents to AIDS or related causes. Sub-Saharan Africa is the most severely affected, accounting for more than 80 percent of those orphaned as a result of AIDS. Without the care of parents or an appointed caregiver, children are likely to face extraordinary risks of malnutrition, poor health, inadequate schooling, migration, homelessness, and abuse. Strengthening existing family and community capacity to assist orphans in Africa should be the first priority. Community support must be coupled with support for education for orphans. Combining local and international responses to deliver protection and services to all orphans and vulnerable children is critical. In addition, saving the lives of parents through access to antiretroviral therapies in resource-poor countries in conjunction with bold support for alleviation of poverty and education must be an integral part of the global response to the orphan crisis in sub-Saharan Africa. PMID- 12748920 TI - The parallel universe of homeless and HIV-positive youth. AB - The HIV/AIDS crisis among teens in this country is alarming, but the rates are even more staggering when these youth are homeless. They tend to live in a world typically considered by those trying to care for them-family, friends, and healthcare providers-as unreachable and hopeless. This article seeks to present "their world" to health professionals in attempts to depict it not as inaccessible, but as a sensitive one that takes great care and support in order for contact to be successful. Their words and those of individuals who have tried to make this connection are used to facilitate the presentation. PMID- 12748921 TI - Health services for homeless adolescents. AB - This prospective review was designed to determine the effectiveness of a broad spectrum health intervention program for homeless and runaway youth. Diagnosis, treatment, and counseling for drug use, sexually transmitted diseases (STDs), and other health issues were provided to all new admissions to a residential care facility during a 7(1/2)-year enrollment. Education was continued during a minimum follow-up period of 9 months based on the program entitled Bright Futures, previously developed and published by the National Center for Education in Maternal and Child Health. Fifty-four percent of the study residents had STDs on admission, and 9% developed new STDs after completing therapy and undergoing counseling. Drug dependence was reduced from 47% to 4%, and 46% achieved full time or part-time employment. Sixty-three percent completed hepatitis B immunization with the 3-dose series. This experience suggests that an organized program of interventions in a residential care facility for homeless teenagers can significantly reduce drug dependence and STDs. PMID- 12748922 TI - Boston HAPPENS Program: HIV-positive, homeless, and at-risk youth can access care through youth-oriented HIV services. AB - The Boston HAPPENS Program is a collaborative network of care consisting of multiservice outreach agencies; community health centers; and hospitals for HIV positive, homeless, and hard-to-reach youth. In four years of data collection, the program served more than 2,000 youth, including 54 HIV-positive youth. The youth were 19.9 +/- 2.9 years old; 64 percent female; 45 percent youth of color; 11 percent gay/lesbian, bisexual, or undecided; and 13 percent homeless or runaway. Homeless youth were much more likely to have been involved with a mental health system (47% vs. 12%, P < 0.001), the criminal justice system (20% vs. 2%, P < 0.001), high-risk sexual behaviors (21% vs. 3%, P < 0.001), and substance abuse (25% vs. 6%, P < 0.001) than were other youth served by the program. Comprehensive networks of care offering a continuum of services and a variety of entry routes and types of care sites are needed to connect under-served youth to health care. Outreach and human immunodeficiency virus (HIV) counseling and testing services can offer important portals of entry into health services for at risk youth. Support services such as outreach, case management, and mental health services are needed to complement medical services by all youth at-risk for contracting HIV. Support services are necessary for the initiation and retention of youth in care so that early case identification and complex treatment regimens can be initiated and tailored to the individual. PMID- 12748923 TI - Appropriate use of nonculture tests for the detection of sexually transmitted diseases in children and adolescents. AB - The introduction of nucleic acid amplification tests (NAATs) has been the most important advance in the diagnosis of sexually transmitted diseases (STDs) since tissue culture replaced inoculation of eggs for culture and isolation of Chlamydia trachomatis from clinical specimens. Because nucleic acid amplification is exquisitely sensitive, theoretically capable of detecting as little as a single gene copy, and highly specific, it offers the opportunity to use noninvasive sampling (ie, urine). This ability in turn means that these tests might be used in nontraditional settings, such as schools and in the field, which may be very helpful in providing services to homeless adolescents. However, none of these tests is approved or recommended by the manufacturers for rectal specimens from adults, and they are not approved for rectogenital specimens from children. In addition to medical implications, the identification of a sexually transmitted disease (STD), especially in a young child, also has legal implications. Because of the legal implications, the highest specificity is needed and is more important than is sensitivity in this situation. Data on the use of these tests in prepubertal children are insufficient to permit their use at this time. What may be appropriate for screening a sexually active adult in an STD clinic may not be appropriate for evaluating a child victim of suspected sexual abuse. Although missing possible sexual abuse is a major concern, the ramifications of a false-positive test for an STD, which can lead to erroneous reports of sexual abuse and possibly unjustified prosecution and incarceration, also must be considered. PMID- 12748924 TI - Biography: Selman A. Waksman, PhD (1888-1973): pioneer in development of antibiotics and Nobel Laureate. AB - Selman Abraham Waksman is regarded today as the foremost authority on soil biology. A Nobel Prize recipient for his work on antibiotics, he also coined the term. Waksman started life in humble surroundings in Priluka, the Ukraine, and later, as a young man, emigrated to the United States, where he later became a renowned scientist. His work on antibiotics has changed the lives of many people, far more than he could possibly have imagined. This brief biography seeks to honor this individual and his contributions to the war against infectious diseases. PMID- 12748926 TI - The adventure of being an editor. PMID- 12748927 TI - Professional practice: Is nursing a profession or a commodity? PMID- 12748928 TI - Research: Altruistic service in research. PMID- 12748929 TI - Public policy: National health care quality initiatives: shaping or critiquing? PMID- 12748930 TI - Nursing and the opportunities of a lifetime. PMID- 12748931 TI - A mentored experience (KO1) in maternal-infant research. AB - Funding mechanisms that require a mentor provide a unique opportunity to implement the research mentoring that is recognized as increasingly important in nursing. Little has been written about how to create and sustain the roles of mentor and principal investigator within a funded arrangement. This article analyzes one research mentoring relationship focused on maternal-infant interaction research and implemented through the Federal KO1 (Mentored Research Scientist Development Award) grant mechanism. Methods used are personal reflections on the 4-year mentorship experience and literature review using the National Library of Medicine database from 1990 to 2001. The mentoring relationship within the National Institutes of Health KO1 award is characterized by the personal commitment, common interests, and long-term professional development that are essential qualities of traditional mentoring but differs in that it is structured by the aims, activities, and length of each award. Effective participation requires organization, focus, and self-direction on the part of the mentored principal investigator and expertise, altruism, and generativity on the part of the mentor. A grant-based mentoring relationship can provide mutual benefits to the researcher and mentor but both must attend to sustaining and enriching the relationship. PMID- 12748933 TI - Portfolio evaluation for professional competence: credentialing in genetics for nurses. AB - The use of professional portfolios, comprised of a wide variety of materials and evidence to profile the scope and depth of a clinician's practice competence, is gaining popularity. The usual methods of showing professional competence via paper and pencil/computerized testing, oral presentations, or performance observations provide a picture of competence at a given point in time based on didactic content recall. Portfolios present an opportunity for presentation of a larger number of competency evaluation points. Although examinations can be validated with psychometrics, providing accuracy and reliability of evaluation of portfolios is a more complicated matter. This article discusses the experiences of the Credentialing Committee of the International Society of Nurses in Genetics as they created and validated the evaluation of professional portfolios to provide a quality credential for nurses in genetics. PMID- 12748932 TI - Faculty practice and roles of staff nurses and clinical faculty in nursing student learning. AB - The perceptions of staff nurses and clinical nurse faculty on the roles they play in nursing student learning were examined. The study also sought to explore how faculty practice status affected these perceived roles. Nursing schools with generic baccalaureate nursing programs in the middle Atlantic region composed the study population. A convenience sample of two schools that expected their undergraduate clinical faculty to engage in faculty practice and two schools that did not expect faculty practice participated in the study. Clinical faculty members (15) and nursing education administrators (4) from each school participated, as well as staff nurses (22) and nursing service administrators (4) from each school's related hospital. Focus groups and individual interviews were conducted. Manifest and latent content analyses were used to examine the issues. Staff nurses who worked with practicing faculty reported fewer instances of role overload, conflict, and ambiguity than when they worked with nonpracticing faculty. Lack of clear communication and delineation of expectations among administrators, staff, faculty, and students were cited as major factors in perceived role problems. Administrators, faculty, and staff nurses viewed high student/faculty ratios as a causative factor in the faculty's inability to meet all teaching objectives with students. PMID- 12748934 TI - Developments in nursing in Ireland: the emergence of a disciplinary discourse. AB - This article presents a conceptualization of nursing as it is presently being constructed in Ireland. Official discourses on nursing are explored and the ways in which these discourses have been further shaped through consultation is highlighted. This is further supported by reference to research examining practicing nurses' discourses. There is some evidence to suggest that Irish nurses may be beginning to internalize many of the nursing concepts articulated at an international and national level over the past 20 years. However, barriers to actualization in practice are identified. Emerging discourses in Ireland suggest the conceptualization of nursing as promoting and maintaining health with nurses as coordinators of care who also engage in the development of therapeutic relationships with patients. The interpersonal aspect of nursing is considered central to the nursing role. PMID- 12748935 TI - Finnish students' perceptions of a visiting professor. AB - Although nurse educators enthusiastically endorse visiting professors, cross school collaboration, cross-cultural teaching, and international faculty exchange programs, no previous studies have documented students' perceptions of visiting faculty. An open-ended questionnaire was used to identify Finnish students' perceptions of a visiting Fulbright professor and how faculty and students can better prepare for educational exchanges. Four themes emerged from the content analysis of responses to the questionnaire: course content, teaching methods, the role of the teacher, and expectations and developmental challenges. Generally, students were satisfied with the course content and teaching methods. They indicated that it was important that the teacher was encouraging and positive and challenged them to speak up in class. Students commented that the visiting professor should perhaps have been more knowledgeable about and therefore able to make applications to Finnish culture, education, and the health care system. Others noted that it was helpful to learn about the American health care system and then to make applications to Finnish culture themselves. Overall, the findings of this project support continuation of visiting professors and continued research on methods of increasing visiting faculty program effectiveness. PMID- 12748936 TI - Recruiting vulnerable populations for research: revisiting the ethical issues. AB - The successful recruitment of participants is essential to the research process. To increase the rate of subject participation, investigators have to rely on help from health care providers who often serve as gatekeepers to potential research study subjects. These well-meaning professionals may limit subject participation in a study involving vulnerable populations under the guise of protecting these individuals from harm. We describe some of the characteristics of vulnerable populations affecting their recruitment into research studies. We examine the ethical conflicts that occur when health care professionals control access to these subjects during recruitment for research studies. Finally, we discuss the implications for practice and research designed to protect the rights of vulnerable populations participating in research. We identify collaboration and dialogue between researchers, health care providers, and members of the potential subject populations as most important in recruiting sufficient numbers of subjects to ensure the scientific merit of the study. PMID- 12748937 TI - Strategies for the enrichment and identification of basic proteins in proteome projects. AB - Two-dimensional gel electrophoresis (2-DE) is currently the method of choice for separating complex mixtures of proteins for visual comparison in proteome analysis. This technology, however, is biased against certain classes of proteins including low abundance and hydrophobic proteins. Proteins with extremely alkaline isoelectric points (pI) are often very poorly represented using 2-DE technology, even when complex mixtures are separated using commercially available pH 6-11 or pH 7-10 immobilized pH gradients. The genome of the human gut pathogen, Helicobacter pylori, is dominated by genes encoding basic proteins, and is therefore a useful model for examining methodology suitable for separating such proteins. H. pylori proteins were separated on pH 6-11 and novel pH 9-12 immobilized pH gradients and 65 protein spots were subjected to matrix-assisted laser desorption/ionization-time of flight mass spectrometry, leading to the identification of 49 unique proteins. No proteins were characterized with a theoretical pI of greater than 10.23. A second approach to examine extremely alkaline proteins (pI > 9.0) utilized a prefractionation isoelectric focusing. Proteins were separated into two fractions using Gradiflow technology, and the extremely basic fraction subjected to both sodium dodecyl sulphate-polyacrylamide gel electrophoresis and liquid chromatography (LC) - tandem mass spectrometry post-tryptic digest, allowing the identification of 17 and 13 proteins, respectively. Gradiflow separations were highly specific for proteins with pI > 9.0, however, a single LC separation only allowed the identification of peptides from highly abundant proteins. These methods and those encompassing multiple LC 'dimensions' may be a useful complement to 2-DE for 'near-to-total' proteome coverage in the alkaline pH range. PMID- 12748938 TI - "Two-in-one" gel for spot matching after two-dimensional electrophoresis. AB - Two-dimensional electrophoresis (2-DE) is one of the most commonly used techniques in proteomic investigations. However, due to the complex interplay of incidence including significant biological sample variations, lengthy steps involved in performing 2-DE as well as exposure time with silver staining, it is sometimes difficult to differentiate authentic differences caused by drug treatment with those artifacts caused by sample variations, running conditions of 2-DE as well as treatment time in silver staining etc. If we can compare pooled samples of control and treatment groups run in a single gel and stained together, we would be more comfortable with our findings. We propose here a low cost and highly effective method for locating differentially expressed proteins before and after drug treatment. This "two-in-one gel" technique might partially solve the problems mentioned above. PMID- 12748939 TI - ProteomeWeb: a web-based interface for the display and interrogation of proteomes. AB - The analysis of proteomes, i.e., the proteins expressed by biological organisms under a given set of conditions at a given time, requires separating complex protein mixtures into discrete protein components, measuring their relative abundances, and identifying the individual protein components. Many types of data are generated during the course of proteome analysis, including graphic images of the protein profiles, flat files containing numeric data, spreadsheets for assimilating numeric data, and relational database tables for integrating data from multiple experiments. As part of a project to describe the proteomes of microbes of interest to the U.S. Department of Energy, a World-Wide Web-based interface has been developed for the display of protein profiles generated by two dimensional gel electrophoresis. The web interface is capable of obtaining protein identifications on the fly, interrogating the quantitative data in the context of available genome sequence information, and relating the proteome data to existing metabolic pathway databases. Analysis of protein expression profiles is expedited, providing the capability to efficiently determine the gene locations for proteins modulated in abundance in response to different growth conditions and to locate the positions of the proteins within specific metabolic pathways. The proteome of the archaeon Methanococcus jannaschii, a microbe for which the complete genome sequence is available, is used to demonstrate the capabilities of this evolving web interface (http://proteomeweb.anl.gov). PMID- 12748940 TI - A strategy for the comparative analysis of serum proteomes for the discovery of biomarkers for hepatocellular carcinoma. AB - Many of the emerging technologies for the global evaluation of gene expression, at both the RNA and protein level, are being applied to the problem of finding biomarkers for human disease progression. These analyses can be made difficult, however, by variation between samples that arises from both technical and nondisease related physiological or genetic causes. In an effort to identify serum polypeptides whose presence or absence correlates with the clinical status of patients at high risk for hepatocellular carcinoma (HCC), we have developed a strategy that helps to focus the analysis on meaningful changes in protein levels above the background of variation. For the current study we divided the patient population into four clinically defined diagnostic groups that represent a generally increasing risk for HCC. Chronic infection with hepatitis B virus (HBV) is a major risk factor for HCC and our groups included patients with no indication of liver disease (healthy), those with inactive chronic HBV, those with active chronic HBV, and patients with a diagnosis of HCC and history of chronic HBV infection. Serum polypeptides from these patients were first analyzed in two-dimensional gels by combining the serum from patients in each of the four groups to generate composite gel profiles. Analysis of these composite gels allowed us to identify two relatively abundant features that were reduced in the HCC group as compared to the healthy group. Tryptic fragment mass fingerprinting identified the features as a carboxy terminal fragment of complement C3 and an isoform of apolipoprotein A1. These two features were examined by two-dimensional gel electrophoresis of serum from each individual in the four groups in order to verify that the inter-group differences seen in composite gels reported changes in abundance for most members of the group, rather than extreme changes for a small fraction of the group. These preliminary studies suggest that a proteomic methodology can be used for the identification of serum biomarkers for HCC and other liver disease. PMID- 12748941 TI - Navigated laser capture microdissection as an alternative to direct histological staining for proteomic analysis of brain samples. AB - Proteomic analysis of the brain is complicated by the need to obtain cells from specific anatomical regions, or nuclei. Laser capture microdissection (LCM) is a technique that is precise enough to dissect single cells within a tissue section, and thus could be useful for isolating specific brain nuclei for analysis. However, we and others have previously demonstrated that histological staining protocols used to guide LCM have detrimental effects on protein separation by two dimensional electrophoresis (2-DE). Here we describe a new LCM method called navigated LCM. This microdissection method uses fixed but unstained tissue as starting material and thus enables us to avoid artifacts induced by tissue staining. By comparing 2-DE results obtained from fixed, unstained LCM brain tissue samples to those obtained from manually dissected samples, we demonstrated that this microdissection process gave similar protein recovery rates and similar resolution of protein spots on 2-DE gels. Moreover, matrix-assisted laser desorption/ionization-time of flight mass spectrometry analysis of selected spots from gels derived from control and fixed, LCM samples revealed that the fixation LCM process had no effect on protein identification. Navigated LCM of tissue sections is therefore a practical and powerful method for performing proteomic studies in specifically defined brain regions. PMID- 12748942 TI - Proteome analysis of secreted proteins during osteoclast differentiation using two different methods: two-dimensional electrophoresis and isotope-coded affinity tags analysis with two-dimensional chromatography. AB - Bone is maintained by two cell types, bone-forming osteoblasts and bone-resorbing osteoclasts. Osteoblasts express two factors, osteoprotegerin and receptor activator of NF-kappaB ligand (RANKL), inhibiting and promoting osteoclast differentiation, respectively. In contrast, modulators of bone resorption expressed by osteoclasts have not been so well studied enough. In the present study, we demonstrate proteome analysis of secreted proteins during osteoclast differentiation to elucidate the molecular mechanism of bone resorption and bone remodeling. To achieve this objective, we chose RAW264.7 cells with RANKL as a homogeneous osteoclast differentiation model and used two methods, two dimensional gel electrophoresis (2-DE) and isotope-coded affinity tags (ICAT) analysis with two-dimensional liquid chromatography. We found 23 spots in 2-DE and 19 proteins in ICAT analysis which were expressed differently during osteoclast differentiation. These two methods gave us closely related but different information about proteins, suggesting they are complementary or at least supplementary methods at present. Cathepsins, osteopontin, legumain, macrophage inflammatory protein-1alpha, and other proteins were observed as up- or down-regulated proteins and are discussed in the context of osteoclast differentiation and bone resorption. In addition to confirming previous observations, this study indicates novel proteins related to osteoclast differentiation which are potential therapeutic targets for the treatment of bone diseases, such as osteoporosis. PMID- 12748943 TI - Cellular and extracellular proteome analysis of Streptococcus mutans grown in a chemostat. AB - The oral pathogen, Streptococcus mutans, was grown under glucose limitation in a chemostat at pH 7.0 and a dilution rate of 0.1 h(-1) to mimic the conditions prevailing in a healthy human oral cavity in between meal times. Solubilized cellular and extracellular proteins were separated by two-dimensional gel electrophoresis (2-DE) and, following tryptic digestion, 421 protein spots analyzed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry or electrospray ionization-tandem mass spectrometry. Analyses of the mass spectral data showed that the proteins matched the translation products of 200 different open reading frames (ORFs) deduced from contigs of the S. mutans UA159 genome and thus represented proteins derived from approximately 11% of the total ORFs of the bacterium. Of the identified proteins, 172 (including one surface protein) were characterized in the cellular fraction, and the remaining 28 (including two surface proteins) were uniquely identified from the culture fluid. The expression and therefore the existence of 30 proteins previously designated as 'hypothetical' or with no known function was confirmed. 2-DE of whole cell lysates revealed only a single intrinsic membrane protein. This is consistent with proteomic analyses of other Gram-positive bacteria where hydrophilic proteins represent the vast majority of those characterized. PMID- 12748944 TI - Dissecting DNA-protein and protein-protein interactions involved in bacterial transcriptional regulation by a sensitive protein array method combining a near infrared fluorescence detection. AB - The protein array methodology is used to study DNA-protein and protein-protein interactions governing gene expression from the Bacillus stearothermophilus PargCo promoter-operator region. Using probes labelled with near-infrared fluorescence dyes with exitation characteristics close to 700 or 800 nm, it is possible to detect signals from proteins (purified or non-purified in Escherichia coli cell extracts) immobilised on a nitrocellulose membrane with a high sensitivity (almost 12 amol of a spotted protein for protein-DNA interactions). Protein array data are confirmed by other methods indicating that molecular interactions of the order 10(-7) M can be monitored with the proposed protein array approach. We show that the PargCo region is a target for binding at least three types of regulatory proteins, ArgR repressors from thermophilic bacteria, the E. coli RNA polymerase alpha subunit and cyclic AMP binding protein CRP. We also demonstrate that the high strength of the PargC promoter is related to an upstream element that binds to the E. coli RNA polymerase alpha subunit. PMID- 12748945 TI - Susceptibility to oxidative stress: proteomic analysis of bronchoalveolar lavage from ozone-sensitive and ozone-resistant strains of mice. AB - Previous studies have shown that the pulmonary response to ozone (O(3)) varies greatly among strains of mice, but the factor(s) and the mechanism(s) that are responsible for this differential susceptibility have not yet been clearly identified. The present study explores the molecular bases for this differential O(3) susceptibility by studying the expression of proteins associated to the epithelial lining fluid (ELF) from two strains of mice, C57BL/6J and the C3H/HeJ, respectively described as O(3)-sensitive and O(3)-resistant. The ELF proteins of these two strains were displayed by two-dimensional gel electrophoresis (2-DE) of bronchoalveolar lavage fluids (BALFs) and the protein patterns obtained with BALF samples of both strains were compared. Two major differences were observed between the BALF 2-DE protein maps obtained from C57BL/6J and C3H/HeJ strains. First, two isoforms of the antioxidant protein 2 (AOP2) were detected in a strain dependent manner: C3J/HeJ possesses only AOP2a (isoelectric point 5.7) and C57BL/6J exhibits only AOP2b (isoelectric point 6.0). Second, the levels of anti inflammatory and immunosuppressive Clara cell protein-16 (CC16) were 1.3 times higher in the BALF from resistant C3H/HeJ than from sensitive C57BL/6 mice. Moreover, two 6 kDa isoforms of CC16 with isoelectric points of 4.9 (CC16a) and 5.2 (CC16b) are detected in both strains. Interestingly, the C57BL/6J strain had a twice decreased level of the acidic isoform of CC16 compared to C3H/HeJ. Our results suggest that AOP2 and CC16 might participate in the protection of the pulmonary tract to O(3)-induced lung injury. The possible differential contribution of specific protein isoforms in the differential susceptibility to oxidative stress is discussed. PMID- 12748946 TI - Serum biomarkers of hepatitis B virus infected liver inflammation: a proteomic study. AB - Hepatitis B virus (HBV), a serious infectious and widespread human pathogen, represents a major health problem worldwide. Chronic HBV infection has a very high risk of evolving into hepatocellular carcinoma. Although considerable progress was made during the recent past, the pathogenesis of HBV infection is still elusive and a definite diagnosis of HBV infected liver information still relies on biopsy histological test. In this report, we used proteomics technology to globally examine HBV infected serum samples aiming at searching for disease associated proteins that can be used as serological biomarkers for diagnosis and/or target proteins for pathogenetic study. By comparing with normal and HBV negative serum samples, we found that at least seven proteins were significantly changed in HBV infected sera. These greatly altered proteins were identified to be haptoglobin beta and alpha2 chain, apolipoprotein A-I and A-IV, alpha1 antitrypsin, transthyretin and DNA topoisomerase IIbeta. The alteration of these proteins is displayed not only in quantity but also in patterns (or specificity), which can be correlated with necroinflammatory scores. In particular, apolipoprotein A-I presents heterogeneous change in expression level with different isoforms and alpha1-antitrypsin produces evidently different fragments implying diverse cleavage pathways. These unique phenomena appear specific to HBV infection. A combination simultaneously considering the quantities and isoforms of these proteins could be a useful serum biomarker (or index) for HBV diagnosis and therapy. PMID- 12748947 TI - Modulation of gene expression by extracellular pH variations in human fibroblasts: a transcriptomic and proteomic study. AB - Homeostasis of the intracellular ionic concentration, in particular that of hydrogen ions, is pivotal to the maintenance of cell function and viability. Nonetheless, pH fluctuations in both the intracellular and the extracellular compartments can occurr during development, in physiological processes and in disease. The influence of pH variations on gene expression has been studied in different model systems, but only for a limited number of genes. We have performed a broad range analysis of the patterns of gene expression in normal human dermal fibroblasts at two different pH values (in the presence and in the absence of serum), with the aim of getting a deeper insight into the regulation of the transcriptional program as a response to a pH change. Using the Affymetrix gene chip system, we found that the expression of 2068 genes (out of 12 565) was modulated by more than two-fold at 24, 48 or 72 h after the shift of the culture medium pH to a more acidic value, stanniocalcin 1 being a remarkable example of a strongly up-regulated gene. Genes displaying a modulated pattern of expression included, among others, cell cycle regulators (consistent with the observation that acidic pH abolishes the growth of fibroblasts in culture) and relevant extracellular matrix (ECM) components. Extracellular matrix protein 2, a protein with a restricted pattern of expression in adult human tissues, was found to be remarkably overexpressed as a consequence of serum starvation. Since ECM components, whose expression is controlled by pH, have been used as targets for biomolecular intervention, we have complemented the Affymetrix analysis with a two-dimensional polyacrylamide gel electrophoresis analysis of proteins which are differentially secreted by fibroblasts at acidic or basic pH. Mass spectrometric analysis of more than 650 protein spots allowed the identification of 170 protein isoforms or fragments, belonging to 40 different proteins. Some proteins were only expressed at basic pH (including, for instance, tetranectin), while others (e.g., agrin) were only detectable at acidic pH. Some of the identified proteins may represent promising candidate targets for biomedical applications, e.g., for antibody-mediated vascular targeting strategies. PMID- 12748948 TI - Proteomic analysis of the reactive phenotype of astrocytes following endothelin-1 exposure. AB - Reactive gliosis is an invariant feature of the pathology of central nervous system (CNS) injury and a major determinant of neuronal survival and regeneration. To begin to understand the alterations in astrocyte protein expression that drive glial changes that occur following injury, we used an established model system (endothelin-1 stimulation of hypertrophy) and proteomic analysis to define a discrete set of differentially expressed proteins and post translational modifications that occur as the astrocytes change from a quiescent to a reactive state. This orchestrated set of changes included proteins involved in cytoskeletal reorganization (caldesmon, calponin, alpha B-crystallin, stathmin, collapsing response mediator protein-2), cell adhesion (vinculin, galectin-1), signal transduction (RACK-1) and astrocyte differentiation (glutamine synthetase). Using proteomic analysis to understand what drives astrocyte expression of these functionally divergent molecules may offer insight into the mechanisms by which astrocytes can exhibit both pro-regenerative and anti-regenerative activities following CNS injury. PMID- 12748949 TI - A reference map of a human pituitary adenoma proteome. AB - In order to compare the proteomes from different cell types of pituitary adenomas for our long-term goal to clarify the molecular mechanisms that participate in the formation of pituitary adenoma, and to detect any tumor-related marker for an "early-stage" diagnosis, the two-dimensional gel electrophoresis (2-DE) reference map of a pituitary adenoma tissue proteome is described here. A vertical, two dimensional (2-D) polyacrylamide gel electrophoresis system and PDQuest image analysis software have been used to provide a high level of between-gel reproducibility and to accurately array each protein expressed in a pituitary adenoma tissue. Mass spectrometry (matrix-assisted laser desorption/ionization time of flight MALDI-TOF and liquid chromatography-electrospray ionization quadrupole-ion trap LC-ESI-Q-IT) and protein databases were used to characterize each protein in the 2-D gel. The results demonstrate that a good reproducibility of the 2-D gel pattern was attained. The position deviation of matched spots among four 2-D gels was 1.95 +/- 0.45 mm in the isoelectric focusing direction, and 1.70 +/- 0.53 mm in the sodium dodecyl sulfate-polyacrylamide gel electrophoresis direction. A total of ca. 1000 protein spots were separated by 2 DE, and 135 protein spots that represent 111 proteins were characterized with mass spectrometry (96 spots for MALDI-TOF, 39 spots for LC-ESI-Q-IT). The characterized proteins include pituitary hormones, cellular signals, enzymes, cellular-defense proteins, cell-structure proteins, transport proteins, etc. Those proteins were located in the cytoplasmic, cellular membrane, mitochondrial, endoplasmic reticulum, nuclear, ribonucleosome, extracellular fractions, or were secreted in plasma, etc. Those identified proteins contribute to a functional profile of the pituitary adenoma proteome. These data will be used to expand the proteome database of the human pituitary, which can be accessed in the website http://www.utmem.edu /proteomics. PMID- 12748950 TI - Proteomic study of human umbilical vein endothelial cells in culture. AB - The endothelium is a single layer of cells lining the inside face of all blood vessels. It constitutes a major metabolic organ which is critically involved in the generation and the regulation of multiple physiological and pathological processes such as coagulation, hemostasis, inflammation, atherosclerosis, angiogenesis and cancerous metastasis dissemination. In order to increase our knowledge about the protein content and the main biological pathways of human vascular endothelial cells, we have undertaken the proteomic analysis of the most explored present endothelial cell model, i.e. primocultures of human umbilical vein endothelial cells (HUVECs). Using low levels of protein loads (~ 30 nug), the association of two-dimensional electrophoresis with matrix-assisted laser desorption/ionization-time of flight mass spectrometry, liquid chromatography tandem mass spectrometry and database interrogations allowed us to identify 53 proteins of suspected endothelial origin in quiescent HUVECs. Beside cytoskeletal proteins such as actin, tubulin, tropomyosin and vimentin, we identified various proteins more especially implicated in cellular motility and plasticity (e.g. cofilin, F-actin capping protein and prefoldin), in regulation of apoptosis and senescence (protease inhibitor 9, glucose related proteins, heat shock proteins, thioredoxin peroxidase, nucleophosmin) as well as other proteins implicated in coagulation (annexin V, high mobility group protein), antigen presentation (valosin containing protein and ubiquitin carboxyl terminal hydrolase isozyme L1) and enzymatic capabilities (glutathione-S-transferase, protein disulfide isomerases, lactate deshydrogenase). The presented annotated 2-D maps of HUVECs will be soon available on the web at http://www. huvec.com. PMID- 12748951 TI - Identification of metastasis-associated proteins by proteomic analysis and functional exploration of interleukin-18 in metastasis. AB - Very little is currently known about mechanisms underlying cancer metastasis. In the present study, metastasis-associated proteomes were separated and identified by comparative proteomic analysis, and the metastasis-related function of candidate protein interleukin-18 (IL-18) was further elucidated. First, a pair of highly and poorly metastatic sublines (termed PLA801D and PLA801C, respectively), originating from the same parental PLA801 cell line, was identified by spontaneous tumorigenicity and metastasis in vivo and characterized by metastatic phenotypes analysis in vitro. Subsequently, a proteomic approach was used to compare the protein expression profiles between PLA801C and PLA801D sublines. Eleven proteins were identified and further verified by one-dimensional Western blotting, Northern blot and/or semiquantitative reverse transciptase polymerase chain reaction analysis. Compared with those in poorly metastatic PLA801C subline, cytokeratin 18, tissue transglutaminase, Rho GDP-dissociation inhibitor 1, tropomyosin, fibroblast type, IL-18 and annexin I were significantly up regulated, while protein disulfide isomerase, heat shock protein 60, peroxiredoxin 1, chlorine intracellular channel protein 1 (CLI1) and creatine kinase, B chain were significantly down-regulated in the highly metastatic PLA801D subline. Intriguingly, all the identified candidate proteins except for CLI1 have been shown to be somehow associated with distinct aspects of tumor metastasis such as cell growth, motility, invasion, adhesion, apoptosis and tumor immunity, etc. Considering that IL-18 was present in highly metastatic PLA801D but absent in poorly metastatic PLA801C, the association of IL-18 with metastasis was further elucidated by introducing IL-18 sense/IL-18 antisense into PLA801C/PLA801D sublines simultaneously. The results demonstrated that ectopically expressed IL-18 promoted cell motility in vitro and down-regulated E cadherin expression of PLA801C transfectants, while IL-18 antisense remarkably decreased cell invasion potency in vitro and notably increased E-cadherin expression of PLA801D transfectants, indicating that IL-18 might play a role in metastasis by inhibiting E-cadherin expression. PMID- 12748952 TI - Proteome analysis of male gametophyte development in rice anthers. AB - We used proteomic analysis to investigate the changing patterns of protein synthesis during pollen development in anthers from rice plants grown under strictly controlled growth conditions. Cytological analysis and external growth measurements such as anther length, auricle distances and days before flowering were used to determine pollen developmental stages. This allowed the collection of synchronous anther materials representing six discrete pollen developmental stages. Proteins were extracted from the anther samples and separated by two dimensional gel electrophoresis to produce proteome maps. The anther proteome maps of different developmental stages were compared and 150 protein spots, which were changed consistently during development, were analysed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry to produce peptide mass fingerprint (PMF) data. Database searches using these PMF data revealed the identities of 40 of the protein spots analyzed. These 40 proteins represent 33 unique gene products. Four protein spots that could not be identified by PMF analysis were analysed by N-terminal microsequencing. Multiple charge-isoforms of vacuolar acid invertase, fructokinase, beta-expansin and profilin were identified. These proteins are closely associated with sugar metabolism, cell elongation and cell expansion, all of which are cell activities that are essential to pollen germination. The existence of multiple isoforms of the same proteins suggests that during the process of pollen development some kind of post translational modification of these proteins occurs. PMID- 12748953 TI - Fragmentation of dihydroxyacetone kinase 1 from Saccharomyces cerevisiae indicates a two-domain structure. AB - Global protein expression in Saccharomyces cerevisiae strains either deleted for both yeast dihydroxyacetone kinases (DAK1 and DAK2) or overexpressing DAK1, was characterized by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). We found protein expression in the double deletion strain to be highly similar to wild-type. In the strain overexpressing Dak1p, nine spots representing fragments of the Dak1p protein in the size range 40-20 kDa and amounting to approximately 30% of total Dak1p, were discovered (native size Dak1p migrates at roughly 60 kDa). Fragments were characterized by matrix-assisted laser desorption/ionization mass spectrometry and electrospray mass spectrometry analyses to represent either the N- or the C-terminal part of the DAK1 protein. Cleavage points, predicted from mass spectrometry and 2-D PAGE data, mapped almost exclusively in the middle region showing low sequence conservation between Dak1p and its closest homologues. We hypothesize that observed Dak1p fragments represent stable structural domains shielded from access by native endoproteases. Furthermore, overexpressing Dak1p with the non-native N-terminus (M)A-, resulted in native size Dak1p and N-terminal Dak1p fragments appearing in two major 2-D PAGE forms of approximately equal size and abundance, but with slightly different isoelectric points. However, when overexpressing Dak1p with the native N-terminus (M)S-, only the more acidic 2-D PAGE form appeared. In the N-terminal acetyltransferase mutant nat1delta, (M)A-Dak1p species were converted into the basic form, arguing twin spots to represent forms with acetylated and deacetylated N-termini. Data thus indicated that (M)A-N-termini, in the Dak1p context, were NatA substrates recognized with 50% lower efficiency than (M)S-N termini. PMID- 12748954 TI - Enrichment of Escherichia coli proteins by column chromatography on reactive dye columns. AB - The reliable identification and analysis of the low abundance proteins expressed by a cell remains a key challenge in the study of cellular proteomes. The analysis of low abundance proteins is a particular problem when using two dimensional gel electrophoresis (2-DE) to resolve the cellular proteins since the technology is unable to display the wide dynamic range of protein levels typically synthesized by cells. We have investigated the use of reactive dye compounds for the enrichment of low abundance cellular proteins prior to analysis by 2-DE. The capacity of reactive dye compounds to bind specific protein species was used as the basis for a general chromatographic tool for protein enrichment. Six reactive dye compounds were investigated in detail for the analysis of Escherichia coli proteins. Whole bacterial cell lysates were passed down columns prepared with the reactive dye compounds. The bound proteins were eluted with 1.5 M NaCl and analyzed by 2-DE. Distinctive protein profiles were observed for the bound proteins recovered from the different reactive dye compounds. Selected proteins enriched by these methods were identified by peptide mass mapping. The enrichment procedure developed using reactive dye compounds were used to investigate acid-induced changes in the proteome of E. coli grown at either pH 7.0 or pH 5.8. Increased levels of expression were observed for a number of proteins (for example, GdhA, PanC, ProC, TkrA, EF-TS and YodA) were observed for E. coli grown at pH 5.8. Five identified proteins (AroG, FabI, GlyA, PurA and EF Tu) showed reduced levels of synthesis for bacteria grown at pH 5.8 compared to pH 7.0. In the case of PanC and FabI the altered expression profiles were only reliably demonstrated using the enrichment protocols. One theme emerging from these data was that the expression of proteins concerned with one-carbon metabolism was perturbed at pH 5.8, which may point to a previously unrecognized affect of low pH stress on the physiology of E. coli cells. We conclude that the prefractionation of cell lysates on reactive dye columns will serve as a valuable generic tool for the analysis of low abundance proteins expressed by both prokaryotic and eukaryotic cells. PMID- 12748955 TI - Analysis of the Shewanella oneidensis proteome by two-dimensional gel electrophoresis under nondenaturing conditions. AB - Proteomes are dynamic, i.e., the protein components of living cells change in response to various stimuli. Protein changes can involve shifts in the abundance of protein components, in the interactions of protein components, and in the activity of protein components. Two-dimensional gel electrophoresis (2-DE) coupled with peptide mass spectrometry is useful for the analysis of relative protein abundance, but the denaturing conditions of classical 2-DE do not allow analysis of protein interactions or protein function. We have developed a nondenaturing 2-DE method that allows analysis of protein interactions and protein functions, as demonstrated in our analysis of the cytosol and crude membrane fractions of the facultative anaerobe Shewanella oneidensis MR-1. Our experiments demonstrate that enzymatic activity is retained under the sample and protein separation methods described, as shown by positive malate dehydrogenase activity results. We have also found protein interactions within both the soluble and membrane fractions. The method described will be useful for the characterization of the functional proteomes of microbial systems. PMID- 12748956 TI - Proteome analysis of the purine stimulon from Lactococcus lactis. AB - A comparative expression proteome analysis was carried out by analyzing differential expression patterns of pulse-labelled proteins on two-dimensional gels under standard conditions and during purine nucleotide starvation, followed by mass spectrometric identification of regulated proteins. Based upon the expression patterns, three stimulons could be identified in Lactococcus lactis subsp. cremoris. The Psu proteins (purine starvation up-regulated) had increased synthesis during purine depletion in a purine auxotroph. Among these proteins were enzymes of the purine biosynthesis pathways (PurE, PurS, PurM, PurL), and enzymes involved in the generation of C1 units (GlyA, Fhs). C1 units are primarily required for purine biosynthesis. Upon analysis of the nucleotide sequence preceding the structural genes for these proteins in the L. lactis IL1403 genome sequence showed that all contained PurBox-Pribnov box structures resembling the PurR activated promoters for the purDEK and purCSQLF operons. Most, and possibly all members of the Psu stimulon are thus members of the PurR regulon. Five Psu proteins could not be identified. The second stimulon, the Psd stimulon (purine starvation decreased), whose members are down-regulated during purine depletion, contained proteins related to protein synthesis (PpsB, EF-TS, trigger factor), or to GTPases (FtsZ, EF-TS); or are involved in energy metabolism (GapB, CcpA). No common regulatory elements could be found for members of this stimulon. Two Psd proteins escaped identification. The last, Dcu (decoynine up-regulated), stimulon contained proteins whose synthesis escaped the severe general depression during inhibition of the GMP synthetase by decoynine. This regulon was comprised of mostly glycolytic enzymes (fructose bisphosphate aldolase, enolase, pyruvate kinase) and translation elongation factors (GTPases: EF-TU, EF-G). Two Dcu proteins could not be identified. Out of 28 proteins subjected to mass spectrometry, 19 could be readily identified despite the fact that only the genome sequence of a strain of L. lactis subsp. lactis was available. The two subspecies share about 85% sequence identity, comparable to the genetic distance between Escherichia coli and Salmonella typhimurium. A success rate of 68% indicates that it may be feasible to perform proteomics based upon genomic sequences of relatives outside the genus. PMID- 12748957 TI - Identification of novel proteins in culture filtrates of Mycobacterium bovis bacillus Calmette-Guerin in the isoelectric point range 6-11. AB - Two-dimensional gel electrophoresis and mass spectrometry were used to identify proteins in the isoelectric point range 6-11 in culture filtrates of Mycobacterium bovis bacillus Calmette-Guerin (BCG). Twelve proteins were identified, three of which had not been described previously. The expression of the identified proteins was comparatively analyzed in culture filtrates of BCG in different growth phases and culture conditions. For some of these proteins, the relative protein abundance in the different culture filtrate preparations was significantly different. The differential expression of the identified proteins is discussed in relation to their putative localization and/or biological function. PMID- 12748961 TI - Sox18 mutations in the ragged mouse alleles ragged-like and opossum. AB - The ragged (Ra) spontaneous mouse mutant is characterised by abnormalities in its coat and cardiovascular system. Four alleles are known and we have previously described mutations in the transcription factor gene Sox18 in the Ra and Ra(J) alleles. We report here Sox18 mutations in the remaining two ragged alleles, opossum (Ra(op)) and ragged-like (Ragl). The single-base deletions cause a C terminal frameshift, abolishing transcriptional trans-activation and impairing interaction with the partner protein MEF2C. The nature of these mutations, together with the near-normal phenotype of Sox18-null mice, suggests that the ragged mutant SOX18 proteins act in a dominant-negative fashion. The four ragged mutants represent an allelic series that reveal SOX18 structure-function relationships and implicate related SOX proteins in cardiovascular and hair follicle development. PMID- 12748962 TI - Sonographic staging of the developmental status of mouse embryos in utero. AB - In mouse developmental studies it is frequently desirable to isolate embryos of a specific age. However, the traditional staging of embryonic development based on postcoital dates often erroneously predicts the embryonic age, resulting in unwarranted sacrifice of the pregnant mother. Here we report a noninvasive way of staging embryonic development in utero. A clinical 14 MHz ultrasound system was employed to assess the morphology and size of developing embryos from embryonic day 7.5 to 18.5. We demonstrate that the developmental age of the mouse embryos can be accurately determined based on the sonographic morphology and size of the embryos. This noninvasive ultrasound application requires no anesthesia of the mice and the entire process of staging can be completed within 5-10 min. Empirically, this approach is applicable to mice of various genetic backgrounds and significantly enhances the efficiency of studying murine embryogenesis. PMID- 12748963 TI - Expression of Sox3 throughout the developing central nervous system is dependent on the combined action of discrete, evolutionarily conserved regulatory elements. AB - SOX3 is one of the earliest neural markers in vertebrates and is thought to play a role in specifying neuronal fate. To investigate the regulation of Sox3 expression we identified cis-regulatory regions in the Sox3 promoter that direct tissue-specific heterologous marker gene expression in transgenic mice. Our results show that an 8.3 kb fragment, comprising 3 kb upstream and 3 kb downstream of the Sox3 transcriptional unit, is sufficient in a lacZ reporter construct to reproduce most aspects of Sox3 expression during CNS development from headfold to midgestation stages. The apparently uniform expression of Sox3 in the neural tube depends, however, on the combined action of distinct regulatory modules within this 8.3 kb region. Each of these gives expression in a subdomain of the complete expression pattern. These are restricted along both the rostral-caudal and dorso-ventral axes and can be quite specific, one element giving expression largely confined to V2 interneuron precursors. We also find that at least some of the regulatory sequences are able to drive expression of the transgene in the CNS Xenopus laevis embryos in a manner that reflects the endogenous Sox3 expression pattern. These results imply that the underlying mechanism regulating early CNS patterning is conserved, despite several substantial differences in neurogenesis between mammals and amphibians. PMID- 12748964 TI - MISSING FLOWERS gene controls axillary meristems initiation in sunflower. AB - The initiation and growth of axillary meristems are fundamental components of plant architecture. Here, we describe the mutant missing flowers (mf) of Helianthus annuus characterized by the lack of axillary shoots. Decapitation experiments and histological analysis indicate that this phenotype is the result of a defect in axillary meristem initiation. In addition to shoot branching, mutation affects floral differentiation. The indeterminate inflorescence of sunflower (capitulum) is formed of a large flat meristem which produces floret primordia in multiple spirals. In wildtype plants a bisecting crease divides each primordium in two distinct bumps that adopt different fate. The peripheral (abaxial) part of the primordium becomes a small leaf-like bract and the adaxial part becomes a flower. In the mf mutant, the formation of flowers at the axil of bracts is precluded. Histological analyses show that in floret primordia of the mutant a clear subdivision in dyads is not established. The primordia progressively bend inside and only large involucral floral bracts are developed. The results suggest that the MISSING FLOWERS gene is essential to provide or perceive an appropriate signal to the initiation of axillary meristems during both vegetative and reproductive phases. PMID- 12748965 TI - Single gene controlling black eyes found from the intercross of two yellow-eyed strains of Heliothis virescens. AB - Black eyes of the moth of Heliothis virescens were controlled by a single, autosomal recessive gene, b. Black-eyed moths were discovered among progeny in an outcross made to test for allelism of two known genes ye, conferring yellow eyes, and yes, conferring yellow eyes and scales. Complementation to the wildtype gray eye color was observed in 686 (99.1%) of the progeny; however, six progeny of one mating exhibited the new phenotype, black eyes. Two black-eyed females mated to a wildtype sibling produced descendents displaying golden eyes, striped eye, purple eyes, white eyes, and "cat's" eyes. No black-eyed progeny were observed in the F2 generation of lines segregating for y, ye, and yes, confirming that black eye was not a combination of those other genes. These newly discovered genes could be useful in basic studies of developmental genetics or in applied transgenesis. PMID- 12748966 TI - Paracrine action of FGF4 during periimplantation development maintains trophectoderm and primitive endoderm. AB - FGF4, a member of the fibroblast growth factor (FGF) family, is absolutely required for periimplantation mouse development, although its precise role at this stage remains unknown. The nature of the defect leading to postimplantation lethality of embryos lacking zygotic FGF4 is unclear and little is known about downstream targets of FGF4-initiated signaling within the various cellular compartments of the blastocyst. Here we report that postimplantation lethality of Fgf4(-/-) embryos is unlikely to reflect strictly mitogenic requirements for FGF4. Rather, our results suggest that FGF4 is required to maintain trophectoderm and primitive endoderm identity at embryonic day 4.5. This result is consistent with the reported in vitro activity of FGF4 in maintaining trophoblast stem cells and with the requirement for receptor tyrosine kinase signaling in primitive endoderm formation. Thus, postimplantation lethality of Fgf4(-/-) embryos likely results from the failure of proper differentiation and function of extraembryonic cell types. PMID- 12748967 TI - Targeted disruption of the heat shock transcription factor (hsf)-2 gene results in increased embryonic lethality, neuronal defects, and reduced spermatogenesis. AB - Heat shock transcription factors (Hsfs) are major transactivators of heat shock protein (Hsp) genes in the response to stress stimuli, but are also thought to be involved in embryonic development and spermatogenesis. Among the three known mammalian Hsfs, Hsf1 is recognized as the most effective transactivator of Hsps in response to thermal challenge, but the role of Hsf2 in regulation of genes under normal or increased stress conditions in vivo remains elusive. To study its physiological function in vivo, we generated mice deficient in hsf2 by gene targeting. We report here that hsf2(-/-) mice exhibit multiple phenotypes, including an increased prenatal lethality occurring between mid-gestation to birth, with fetal death probably due to central nervous system defects including collapse of the lateral ventricles and ventricular hemorrhages. Approximately 30% of hsf2(-/-) animals surviving to adulthood exhibited brain abnormalities characterized by marked dilation of the third and lateral ventricles. In addition, disruption of hsf2 resulted in reduced female fertility; however, despite ubiquitous expression in the testes and markedly reduced testis size and sperm count, only a small reduction in fertility was apparent in hsf2(-/-) male mice. Immunoblotting and gene expression microarray analysis of hsf2(-/-) embryos did not reveal reduced Hsp expression levels, indicating that the defects observed in hsf2(-/-) embryos may not result from disruption of Hsp expression. These findings suggest that hsf2 has a major function in controlling expression of genes important for embryonic development and maintenance of sperm production. PMID- 12748968 TI - Large-scale characterization of genes specific to the larval nervous system in the ascidian Ciona intestinalis. AB - The central and peripheral nervous systems (CNS and PNS) of the ascidian tadpole larva are comparatively simple, consisting of only about 350 cells. However, studies of the expression of neural patterning genes have demonstrated overall similarity between the ascidian CNS and the vertebrate CNS, suggesting that the ascidian CNS is sufficiently complex to be relevant to those of vertebrates. Recent progress in the Ciona intestinalis genome project and cDNA project together with considerable EST information has made Ciona an ideal model for investigating molecular mechanisms underlying the formation and function of the chordate nervous system. Here, we characterized 56 genes specific to the nervous system by determining their full-length cDNA sequences and confirming their spatial expression patterns. These genes included those that function in the nervous systems of other animals, especially those involved in photoreceptor mediated signaling and neurotransmitter release. Thus, the nervous system specific genes in Ciona larvae will provide not only probes for determining their function but also clues for exploring the complex network of nervous system specific genes. PMID- 12748969 TI - Experience with breast cancer, pre-screening perceived susceptibility and the psychological impact of screening. AB - This prospective study examined whether the psychological impact of organized mammography screening is influenced by women's pre-existing experience with breast cancer and perceived susceptibility (PS) to the disease. From a target population of 16,886, a random sample of women with a normal screening finding and all women with a false positive or a benign biopsy finding were included (N=1942). Data were collected with postal questionnaires 1-month before screening invitation and 2 and 12 months after screening. Response rate was 63% at baseline; 86, and 80% of the baseline participants responded to the follow-ups. Psychological impact was measured as anxiety (STAI-S), depression (BDI), health related concerns (IAS), and breast cancer-specific beliefs and concerns. Data was analyzed with repeated measures analyses of variance, with estimates of effect size based on Eta-squared. Women with breast cancer experience had higher risk perception already before screening invitation; after screening they were also more distressed. Women with high PS were more distressed than women with low PS also at pre-invitation. The distress was not alleviated by screening, but instead remained even after normal mammograms. Experience and PS did not influence responses to different screening findings. Of the finding groups, false positives experienced most adverse effects: their risk perception increased and they reported most post-screening breast cancer-specific concerns. Furthermore, they became more frequent in breast self-examination (BSE) despite a simultaneous decrease in BSE self-efficacy. Our findings suggest that women with high PS and women with false positive screening finding may need individualized counseling and follow-up as much as women with a family history of breast cancer. Besides medical risk factors, women's own perceptions of susceptibility should be discussed during the screening process. PMID- 12748970 TI - An interactive process model of psychosocial support needs for women living with breast cancer. AB - Psychosocial support is acknowledged as an important aspect of the care and recovery process for women diagnosed with breast cancer. To develop an understanding of support needs, a series of focus groups were conducted with a total of 80 Australian women living with breast cancer. The psychosocial needs identified in discussion were summarised into four main categories, Organisation of Care, Sense of Control, Validation of Experience and Feeling of Reassurance. From these themes an interactive model of psychosocial support needs was developed. As a process model it demonstrates that the provision of effective support is, necessarily, an integrated, not a piecemeal, process. The categories of support are broad, and are made more inclusive through interactions with each other, providing room to accommodate individual needs and styles of adjustment. The model is not esoteric or complicated - it is an accessible overview of the process for both client and helper. PMID- 12748971 TI - The Cancer Coping Questionnaire: a self-rating scale for measuring the impact of adjuvant psychological therapy on coping behaviour. AB - The Cancer Coping Questionnaire is a brief, self-rating scale designed to measure coping strategies taught in Adjuvant Psychological Therapy. This paper describes the development of the 21 item Cancer Coping Questionnaire (CCQ) in a sample of 201 patients with mixed cancers. The construct validity and reliability of the instrument are reported from work on 3 samples (a mixed cancer group, n=42; women with breast cancer, n=50; and a group of patients referred for psychological help, n=48). The CCQ showed very good internal reliability and test-retest reliability. As hypothesised cancer patients with more psychological morbidity demonstrated lower CCQ scores, and the CCQ correlated with measures of adjustment to cancer. Compared with an established coping inventory (the Coping Responses Indices; CRI) the CCQs overall individual scale (items 1-14) assessed similar coping areas, particularly in relation to the CRIs foci of coping. The CCQ correlated with Active Behavioural Coping methods on the CRI. The study did not demonstrate sufficiently consistent results concerning the Interpersonal Scale of the CCQ to confirm its validity. Further psychometric work is needed, but the study demonstrated the reliability and validity of the CCQ, supporting the view that change in CCQ scores with cognitive therapy indicates improvement in coping. PMID- 12748972 TI - Construct validity of newly developed quality of life assessment instrument for child and adolescent cancer patients in Taiwan. AB - The purpose of this study was to test the convergent, discriminant, and clinical validity of the Quality of Life in Childhood Cancer (QOLCC) instrument for measuring the quality of life of Taiwanese children who suffer from cancer. In total, 160 patients were recruited for the study, including 105 male and 55 female. Overall, QOLCC consisted of generic measure and disease-specific domains to assess the Quality of Life (QOL) for children treated for cancer. The QOLCC is a symptom or problem-based questionnaire with the conceptual framework that health-related problems can be solved from both a biomedical perspective (e.g. changing the patient's medical treatment can enhance the QOL) and from a biobehavioral perspective (e.g. problem solving on a daily basis). This QOLCC, which can be administered in 15 min, is the first documented measure of the QOL administered directly to Taiwanese Children. It demonstrates acceptable psychometric properties. Application of the QOLCC to Taiwanese children with cancer produced encouraging results, validation from a larger independent parent population is still necessary. PMID- 12748973 TI - The group psychotherapy and home-based physical exercise (group-hope) trial in cancer survivors: physical fitness and quality of life outcomes. AB - Physical exercise has been shown to enhance quality of life (QOL) in cancer survivors using pretest-posttest designs and compared to usual care (i.e. no intervention). In the present study, we conducted a randomized controlled trial to determine if exercise could improve QOL in cancer survivors beyond the known benefits of group psychotherapy (GP). We matched 22 GP classes (N=108) on content and then randomly assigned 11 (n=48) to GP alone and 11 (n=60) to GP plus home based, moderate-intensity exercise (GP+EX). Participants completed a physical fitness test and QOL measures (e.g. Functional Assessment of Cancer Therapy scales) at the beginning and end of GP classes (about 10 weeks). We had excellent recruitment (81%), retention (89%), and adherence (84%) rates and a modest contamination (22%) rate. Using intention-to-treat repeated measures analyses of variance, we found significant Time by Condition interactions for functional well being, fatigue, and sum of skinfolds. We also found borderline significant interactions for physical well-being, satisfaction with life, and flexibility. All interactions favored the GP+EX condition. We conclude that a home-based, moderate intensity exercise program may im-prove QOL in cancer survivors beyond the benefits of GP, particularly in relation to physical and functional well being. PMID- 12748974 TI - Doctors and their patients: a context for understanding the wish to hasten death. AB - There is a paucity of research that has directly examined the role of the health professional in dealing with a terminally ill patient's wish to hasten death (WTHD) and the implications of this for the support and services needed in the care for a dying patient. Themes to emerge from a qualitative analysis of interviews conducted on doctors (n=24) involved in the treatment and care of terminally ill patients were (i). the doctors' experiences in caring for their patients (including themes of emotional demands/expectations, the duration of illness, and the availability of palliative care services); (ii). the doctors' perception of the care provided to their respective patients (comprising themes concerning satisfaction with the care for physical symptoms, for emotional symptoms, or overall care); (iii). the doctors' attitudes to euthanasia and (iv). the doctors' perception of their patients' views/beliefs regarding euthanasia and hastened death. When responses were categorised according to the patients' level of a WTHD, the theme concerning the prolonged nature of the patients' illnesses was prominent in the doctor group who had patients with the highest WTHD, whereas there was only a minority of responses concerning support from palliative care services and satisfaction with the level of emotional care in this group.This exploratory study presents a set of descriptive findings identifying themes among a small group of doctors who have been involved in the care of terminally ill cancer patients, to investigate factors that may be associated with the WTHD among these patients. The pattern of findings suggest that research investigating the doctor-patient interaction in this setting may add to our understanding of the problems (for patients and their doctors) that underpins the wish to hasten death in the terminally ill. PMID- 12748975 TI - Research interests in the field of behavioral, psychosocial, and policy cancer research. AB - The growth of research in behavioral, psychosocial, and policy aspects of cancer is quite evident through examination of the literature. Although it is necessary to recognize the importance of past achievements in these areas, it is equally essential to identify the current interests and future areas that will form a new research agenda. The present study is the first attempt to shed light on these issues by surveying a group of behavioral, psychosocial and policy researchers from 1997 (n=714) and 2002 (n=1102). Questions were posed that dealt with current and future research interests. Results indicate a stable core of research interests in the field, as well as changing trends in interest that may affect the direction of future research. Prevention and detection, psychosocial issues, and quality of life were clearly the most recognized research interests. Emerging areas of research interest seem to be special populations and health care delivery. Examination of these findings may help with research planning, funding allocation, management, beginning researcher education, and practice. PMID- 12748976 TI - Reporter gene assays and their applications to bioassays of natural products. AB - Medicinal plants represent precious resources from which bioactive compounds can be isolated and developed into invaluable therapeutic agents. With the advent of modern drug discovery technologies such as combinatorial chemistry and high throughput drug screening platforms, there is an increasing interest in utilizing medicinal plants as a source of drug leads. A wide spectrum of bioassays can be employed for the detection of bioactivity in extracts, fractions, as well as purified compounds of herbal origin. Amongst the different types of bioassays, reporter gene assays are highly versatile and reliable. The present review provides an overview of the most popular reporter genes in terms of their basic methodology, capacities and limitations. The different types of intracellular and extracellular reporter gene products and their potential applications in bioassays of natural products are also discussed. PMID- 12748977 TI - Screening of 25 compounds isolated from Phyllanthus species for anti-human hepatitis B virus in vitro. AB - Using an HBV-producing cell line and inhibition of the expression of the HBsAg and HBeAg as antiviral indicators, a study was conducted on 25 compounds isolated from four Phyllanthus (Euphorbiaceae) plants, including P. amarus Schum. & Thonn., P. multi florus Willd., P. tenellus Roxb. and P. virgatus Forst. f. It was found that niranthin (1), nirtetralin (3), hinokinin (5) and geraniin (13) at the non-cytotoxic concentration of 50 micro m, suppressed effectively both HBsAg and HBeAg expression, with the highest inhibition at 74.3%, 45.3%; 69.6%, 33.9%; 68.1%, 52.3%; 32.1%, 46.6%, respectively. Of these, niranthin (1) showed the best anti-HBsAg activity, while the most potent anti-HBeAg activity was observed with hinokinin (5). PMID- 12748979 TI - Screening of Tanzanian plant extracts for their potential inhibitory effect on P glycoprotein mediated efflux. AB - For years, many efforts have been made to discover new drugs using plants as natural screening libraries. In this study, extracts of 43 Tanzanian medicinal plants were screened for their potential inhibitory effect on P-gp, using the secretory transport of Cyclosporin A (CsA) in the Caco-2 system as a measure of the functionality of P-gp efflux. Two out of these 43 plant extracts (extracts of Annickia kummeriae and Acacia nilotica) appeared to have a modulatory effect on P gp related efflux carriers. In presence of the extract of Annickia kummeriae, a concentration dependent decrease on the polarity in transport of CsA was observed; the inhibitory effect of this extract on P-gp was comparable to that of valspodar, a known P-gp inhibiting agent. The exact nature of the active components of these botanicals remains to be identified. PMID- 12748978 TI - Short-term control of capsaicin on blood and oxidative stress of rats in vivo. AB - Capsaicin (8-methyl-n-vanillyl-6-nonenamide), a pungent component found in red pepper can induce body heat and possibly enhance blood flow as well as increase energy expenditure, and prevent oxidative stress. Male Wistar rats were divided into vehicle, 1 mg/kg body weight capsaicin and 3 mg/kg body weight capsaicin groups. Samples were taken from the animals on day 1 of i.p. treatment with capsaicin and on 3 consecutive days of i.p. treatment with capsaicin. Our investigation demonstrated that blood flow measurements in rats was negatively correlated with LDL after treatment with capsaicin. Although capsaicin did not show a noticeable effect on the serum total cholesterol level, LDL decreased while HDL and triglyceride increased in rats treated with 3 mg/kg capsaicin for 3 days. The antioxidant effect of capsaicin was not shown when the rats were treated with 1 mg/kg body weight capsaicin. However, rats treated with 3 mg/kg body weight capsaicin for 3 days showed a reduction of oxidative stress measured as malondialdehyde in the liver, lung, kidney and muscle. Liver glycogen was found to decrease after 3 days treatment with 3 mg/kg body weight capsaicin. From this study, it is hypothesized that capsaicin can be a potent antioxidant and aid in lowering LDL even when consumed for a short period. PMID- 12748980 TI - In vitro and in vivo antioxidant effects of mustard leaf (Brassica juncea). AB - To investigate the antioxidant activity of mustard leaf (Brassica juncea), we prepared four fractions (CH(2)Cl(2), EtOAc, BuOH and H(2)O fractions) and examined their radical scavenging activities in vitro and in vivo. Based on the in vitro results of spin trapping and 1,1-diphenyl-2-picrylhydrazyl radical, we carried out an in vivo study with the BuOH fraction to investigate its effect on oxidative stress in rats with streptozotocin-induced diabetes. We found that in comparison with untreated diabetic control rats, oral administration of the BuOH fraction (100 or 200 mg/kg body weight/day for 10 days) induced a significant decrease in serum glucose and glycosylated protein, which is glycosylated with hemoglobin as an indicator of oxidative stress. Moreover, administration of the BuOH fraction also effectively reduced the serum superoxide and nitrite/nitrate levels. Furthermore, the levels of thiobarbituric acid-reactive substances in serum and liver were also significantly lower than in the control group. These results indicate that the BuOH fraction of mustard leaf controls glucose metabolism and reduces lipid peroxidation as well as the level of oxygen radicals, ameliorating the damage caused by oxidative stress in diabetes. PMID- 12748981 TI - Cytotoxic activities of some Greek Labiatae herbs. AB - Nineteen methanolic crude plant extracts of Labiatae family, collected in Greece from different locations, were evaluated for cytotoxic activity against brine shrimps and three human cancer cell lines along with a normal mouse cells as a control cell line. In the brine shrimp lethality test, Mentha pulegium was the only sample found to be active with an LC(50) value 347.3 micro g/ml, while all remaining samples had LC(50) values greater than 1000 micro g/ml. In case of Caco 2 and HepG2 cell lines, only one sample, namely Thymus parnassicus Halacsy, was active with LC(50) values 44.6 and 50.3 micro g/ml respectively, while against MCF-7 cell line, two samples, namely, Clinopodium vulgare L. (LC(50): 60.4 micro g/ml), and Thymus parnassicus Halacsy (LC(50): 54.7 micro g/ml), were found active. PMID- 12748982 TI - Powdered green tea has antilipogenic effect on Zucker rats fed a high-fat diet. AB - The effect of powdered green tea on the lipid metabolism was studied in male Zucker rats fed a 50% sucrose diet containing 15% butter. The oral treatment of 130 mg powdered green tea per day depressed body weight increase and various adipose tissue weights but food intake was unaffected. The treatment also slightly increased the plasma triglycerides. These effects were likely mediated by the inhibition of lipogenesis in the adipose tissues. The experimental group had a higher concentration of liver total lipid, triglycerides and plasma protein, and lower liver weights than the controls. This may be due to lipid deposition in the liver because of the reduction in adipose tissue weights. It was found that powdered green tea lowered the plasma total cholesterol but liver total cholesterol was unaffected. The results indicate that the hypocholesterolemic activity of powdered green tea might be due to the inhibition of the synthesis of cholesterol in the liver. PMID- 12748983 TI - In vitro peroxynitrite scavenging activity of diarylheptanoids from Curcuma longa. AB - Peroxynitrite is a cytotoxic intermediate produced by the reaction between the superoxide anion (O(2)) and nitric oxide (NO). The aim of this study was to investigate the scavenging effects of Curcuma longa L. on authentic peroxynitrite, and further studies are planned that will attempt to identify the active principles from the active fractions. The methanolic extract of C. longa showed 50% scavenging activity (IC(50)) at concentration of 1.7 +/- 0.08 micro g/ml, and was thus fractionated with several solvents. The peroxynitrite scavenging activity potential of the individual fraction was in the order of ethyl acetate > dichloromethane > water fraction. The ethyl acetate soluble fraction exhibiting strong scavenging activity was further purified by repeated silica gel column chromatography. Peroxynitrite scavenging diarylheptanoids, curcumin I (1), curcumin II (2), and curcumin III (3) were isolated as active principles. Compounds 1-3 showed the peroxynitrite scavenging activities with IC(50) values of 4.0 +/- 0.04, 6.4 +/- 0.30, and 29.7 +/- 1.29 micro M, respectively. Penicillamine as positive control exhibited IC(50) value of 2.38 +/ 0.34 micro M. The structure-activity relationship of diarylheptanoids on peroxynitrite was also discussed. PMID- 12748984 TI - Inhibitory activity of plant extracts on nitric oxide synthesis in LPS-activated macrophages. AB - Nitric oxide (NO) produced in large amounts by inducible nitric oxide synthase (iNOS) is known to be responsible for the vasodilation and hypotension observed in septic shock and inflammation. Inhibitors of iNOS, thus, may be useful candidates for the treatment of inflammatory diseases accompanied by overproduction of NO. We prepared alcoholic extracts of woody plants and screened the inhibitory activity of NO production in lipopolysaccharide (LPS)-activated macrophages after the treatment of these extracts. Among 83 kinds of plant extracts, 23 kinds of extracts showed potent inhibitory activity of NO production above 60% at the concentration of 80 micro g/ml. Some of potent extracts showed dose dependent inhibition of NO production of LPS-activated macrophages at the concentration of 80, 40, 20 micro g/ml. Especially, Artemisia iwayomogi, Machilus thunbergii, Populus davidiana and Populus maximowiczii showed the most potent inhibition (above 70%) at the concentration of 40 micro g/ml. Inhibitory activity of NO production was concentrated to nonpolar solvent fractions (ethyl ether and/or ethyl acetate soluble fractions) of Artemisia iwayomogi, Machilus thunbergii and Morus bombycis. These plants are promising candidates for the study of the activity-guided purification of active compounds and would be useful for the treatment of inflammatory diseases and endotoxemia accompanying overproduction of NO. PMID- 12748985 TI - Pilot study on the clinical effects of dietary supplementation with Enzogenol, a flavonoid extract of pine bark and vitamin C. AB - Flavonoids are naturally occurring plant compounds with established in vitro antioxidant properties and potential cardioprotective effects. We carried out a 12-week pilot study on the effects of dietary supplementation with an extract of bioflavonoids prepared from the bark of Pinus radiata trees [Enzogenol] containing added vitamin C. Data was collected from 24 healthy subjects aged between 55-75 years at baseline and at 6 and 12 weeks and included, routine biochemical and haematological indices, and anthropometric, blood pressure, forearm blood flow and haemorheological measurements. Enzogenol supplementation at a dosage of 480 mg/day of pine bark extract and 240 mg/day vitamin C did not result in changes in any biochemical or haematological indice and was associated with a significant reduction in the means of body weight, percentage body fat, systolic blood pressure and plasma viscosity. Basal and hyperaemic blood fl ow in forearm resistance vessels measured by plethysmography increased significantly during the study. The findings of this pilot study indicate that dietary supplementation with Enzogenol is safe and well tolerated and is associated with a number of beneficial effects on a range of established cardiovascular risk factors. These changes need to be validated by a placebo-controlled study but are consistent with other studies that have reported beneficial clinical effects following supplementation with bioflavonoids. PMID- 12748986 TI - Biological activity of persimmon (Diospyros kaki) peel extracts. AB - Fractionated extracts of persimmon (Diospyros kaki) peels were studied for cytotoxic activity, multidrug resistance (MDR) reversal activity, anti-human immunodeficiency virus (HIV) activity and anti-Helicobacter pylori (H. pylori) activity. The potent cytotoxic activity against human oral squamous cell carcinoma cells (HSC-2) and human submandibular gland tumor (HSG) cells was found in the acetone fractions (A4 and A5) with IC(50) ranging from 21 to 59 micro g/mL. However, the cytotoxic activity was not correlated with the radical intensity of the fractions. Three 70% MeOH extract fractions (70M2-4) produced radical and efficiently scavenged the O(2)(-) produced by hypoxanthine and xanthine oxidase reaction. All of the fractions tested were not effective for anti-H. pylori and anti-HIV. Fractions H3 and H4 of hexane extract, and M2 and M3 of MeOH extract showed a remarkable MDR reversal activity comparable with that of (+/-)-verapamil (a positive control). These results indicate the therapeutic value of persimmon peel extracts as potential antitumor and MDR-reversing agents. PMID- 12748987 TI - Biological activity of some naturally occurring resins, gums and pigments against in vitro LDL oxidation. AB - Naturally occurring gums and resins with beneficial pharmaceutical and nutraceutical properties were tested for their possible protective effect against copper-induced LDL oxidation in vitro. Chiosmastic gum (CMG) (Pistacia lentiscus var. Chia resin) was the most effective in protecting human LDL from oxidation. The minimum and maximum doses for the saturation phenomena of inhibition of LDL oxidation were 2.5 mg and 50 mg CMG (75.3% and 99.9%, respectively). The methanol/water extract of CMG was the most effective compared with other solvent combinations. CMG when fractionated in order to determine a structure-activity relationship showed that the total mastic essential oil, collofonium-like residue and acidic fractions of CMG exhibited a high protective activity ranging from 65.0% to 77.8%. The other natural gums and resins (CMG resin 'liquid collection', P. terebinthus var. Chia resin, dammar resin, acacia gum, tragacanth gum, storax gum) also tested as above, showed 27.0%-78.8% of the maximum LDL protection. The other naturally occurring substances, i.e. triterpenes (amyrin, oleanolic acid, ursolic acid, lupeol, 18-a-glycyrrhetinic acid) and hydroxynaphthoquinones (naphthazarin, shikonin and alkannin) showed 53.5%-78.8% and 27.0%-64.1% LDL protective activity, respectively. The combination effects (68.7%-76.2% LDL protection) of ursolic-, oleanolic- and ursodeoxycholic- acids were almost equal to the effect (75.3%) of the CMG extract in comparable doses. PMID- 12748988 TI - Antifertility effects of Ricinus communis (Linn) on rats. AB - The antifertility effects of 50% ethanol extracts of Ricinus communis have been studied in male rats. There was a drastic reduction in the epididymal sperm counts. Alteration in the motility, mode of movement and morphology of the sperms were observed. Reductions in the fructose and testosterone levels were suggestive of reduced reproductive performance. Reversibility tests showed that the antifertility effect of Ricinus communis was completely reversible on withdrawal of the drug. The ethanol extracts of Ricinus communis did not cause any hepatotoxicity since the hepatic GOT and GPT levels were unaltered. PMID- 12748989 TI - Carpobrotus edulis methanol extract inhibits the MDR efflux pumps, enhances killing of phagocytosed S. aureus and promotes immune modulation. AB - Although alkaloids from the family Aizoaceae have anticancer activity, species of this family have received little attention. Because these alkaloids also exhibit properties normally associated with compounds that have activity at the level of the plasma membrane, a methanol extract of Carpobrotus edulis, a common plant found along the Portuguese coast, was studied for properties normally associated with plasma membrane active compounds. The results of this study show that the extract is non-toxic at concentrations that inhibit a verapamil sensitive efflux pump of L5178 mouse T cell lymphoma cell line thereby rendering these multi-drug resistant cells susceptible to anticancer drugs. These non-toxic concentrations also prime THP-1 human monocyte-derived macrophages to kill ingested Staphylococcus aureus and to promote the release of lymphokines associated with cellular immune functions. The extract also induces the proliferation of THP-1 cells within 1 day of exposure to quantities normally associated with phytohaemagglutinin. The potential role of the compound(s) isolated from this plant in cancer biology is intriguing and is currently under investigation. It is supposed that the resistance modifier and immunomodulatory effect of this plant extract can be exploited in the experimental chemotherapy of cancer and bacterial or viral infections. PMID- 12748990 TI - Cytotoxic constituents of Brachistus stramoniifolius. AB - Phytochemical investigation of Brachistus stramoniifolius (Kunth) Miers (formerly Witheringia stramoniifolia Kunth) (Solanaceae) was initiated following primary biological screening and in view of the absence of prior phytochemical studies conducted on this species. Fractionation of an ethyl acetate extract from the roots of the plant, guided by in vitro cytotoxic activity using cultured KB (human nasopharyngeal carcinoma) cells, led to the isolation of three known compounds of the 13,14-seco-16,24-cyclosteroid type, physalins B (1), F (2) and H (3). Their structures were characterized by comparison of their physical and spectral data to published values, with 1D- and 2D-NMR experiments being performed to assure unambiguous resonance assignments. Biological evaluation of these three compounds against a panel of human and murine cancer cell lines demonstrated their broad cytotoxic activity. PMID- 12748992 TI - Adaptogenic activity of a novel withanolide-free aqueous fraction from the roots of Withania somnifera Dun. (Part II). AB - In the Indian traditional system of medicine Withania somnifera Dun. is widely regarded as the Indian Ginseng. A new withanolide-free hydrosoluble fraction was isolated from the roots of Withania somnifera Dun. and was evaluated for putative antistress activity against a battery of tests to delineate the activity of this fraction. The latter fraction exhibited significant antistress activity in a dose related manner (Singh et al., 2001) and was further studied against chemical and physical induced stress in rats and mice. The extract of Withania somnifera root (a commercial preparation available locally) was also used to compare the results. A preliminary acute toxicity study in mice showed a good margin of safety with a high therapeutic index. PMID- 12748991 TI - Plasma lipoproteins in transport of silibinin, an antioxidant flavonolignan from Silybum marianum. AB - To assess the role of plasma lipoproteins in the transport of silibinin, an antioxidant flavonolignan, (125)I-labelled silibinin ((125)I-SB) administered perorally to the rat was used. The plasma (125)I-SB derived radioactivity was distributed among plasma lipoproteins according to their lipophilicity (TAG-rich lipoproteins 30-40% > LDL 15% > HDL 5%), and in the fraction of d > 1.215 containing albumin and other proteins a minority amount of radioactivity was found. Administration of (125)I-SB in a complex with phosphatidylcholine resulted in proportionally higher radioactivities in all fractions as well as in tissues. Dietary olive oil had a slightly decreasing effect on plasma concentrations of silibinin measured by HPLC as well as on (125)I-SB derived radioactivity in plasma and liver. In the TAG-rich lipoprotein fraction and HDL no effects of olive oil on the levels of (125)I-SB derived radioactivities were observed, however, at a 30 min interval the levels of (125)I-SB derived radioactivity in LDL and the heart were significantly decreased in the olive oil group. These results suggest that (i) silibinin is not resorbed by the chylomicron pathway, and (ii) the endogenous lipoprotein pathway VLDL --> LDL may play a role in the transport of silibinin from the liver to the extrahepatic tissues concurrently facilitating the lipoprotein antioxidant influence of silibinin. PMID- 12748993 TI - Reducing effect of Salvia miltiorrhiza extracts on alcohol intake: influence of vehicle. AB - A previous study demonstrated that an extract of Salvia miltiorrhiza, a medicinal herb highly valued in Chinese folk medicine for the treatment of different pathologies, including insomnia, was capable of reducing voluntary alcohol intake in selectively bred Sardinian alcohol-preferring (sP) rats. The present study was designed to evaluate the suitability of different emulsifying, suspending agents and solvents as vehicles through which Salvia miltiorrhiza extracts can exert their reducing effect on alcohol intake. A single dose (100 mg/kg) of a standardised extract of Salvia miltiorrhiza was dissolved in either pure Polysorbate 80, arachis oil, PEG 400, or Polyoxyl 35 castor oil, or suspended in 0.5% CMC in water, and administered acutely by gavage to sP rats. A significant and specific reduction in alcohol intake was recorded only in rats treated with the combination of Polysorbate 80 plus the Salvia miltiorrhiza extract. A further experiment demonstrated that the ability of the combination of Polysorbate 80 in water plus the Salvia miltiorrhiza extract to decrease alcohol intake was dependent upon the concentration of Polysorbate 80. The results of the present study demonstrate that Polysorbate 80 is a proper vehicle for unravelling the reducing effect of Salvia miltiorrhiza extracts on alcohol intake. The ability of Polysorbate 80 to form micelles with the active ingredient(s) of the Salvia miltiorrhiza may explain these results. They may also offer relevant information for pharmaceutical preparation of Salvia miltiorrhiza extract to be used in future clinical trials. PMID- 12748994 TI - Effect of Choto-san, a Kampo medicine, on impairment of passive avoidance performance in senescence accelerated mouse (SAM). AB - Effect of Choto-san (TJ-47), a Kampo medicine, on impairment of learning performance was evaluated by means of a step-through passive avoidance task in SAMP8 mice, a senescence-prone substrain. Tokishakuyaku-san (TJ-23), another Kampo medicine, was also employed for comparison. SAMP8 mice at the age of 10-12 months showed a poorer passive avoidance response than SAMR1 mice, a senescence resistant substrain, in the memory-retention test, but not in the memory acquisition test. Improved response in the memory-retention test was observed in SAMP8 mice treated with TJ-47, and almost equal degree of improvement was also observed after treatment with TJ-23. These results suggest that a long-term administration of Choto-san could improve to some extent the impairment of memory caused by aging. PMID- 12748995 TI - Inhibitory effects of aromatic herbs on lipid peroxidation and protein oxidative modification by copper. AB - Aromatic herbs have been used as carminatives. Oxygen free radicals are generated in ischaemia/reperfusion injury in the stomach, and induce lipid peroxidation or protein oxidative modification. Several aromatic herbs were shown to have inhibitory effects on the generation of oxygen free radicals. It was shown that several aromatic herbs, Caryophylli Flos, Cinnamomi Cortex, Foeniculi Fructus and Zedoariae Rhizoma, have inhibitory effects on lipid peroxidation or protein oxidative modification by copper. PMID- 12748996 TI - Inhibitory activity on lipid peroxidation of extracts from marine brown alga. AB - Several extracts from marine brown alga Sargassum micracanthum (Kuetzing) Endlicher were screened for their inhibitory activity on lipid peroxidation. In an in vitro study, methanol extract (Sm-M), chloroform/ methanol (3:1) extract and ethyl acetate fraction of Sm-M inhibited lipid peroxidation in rat liver homogenates. The IC(50) values were 0.70, 0.70 and 0.37 micro g/mL, respectively. These inhibitions were stronger than vitamin C and E. These extracts showed reductive activity on DPPH, the IC(50) values were 34, 37 and 11 micro g/mL, respectively. In an in vivo study, Sm-M had the effect on CCl4 induced liver injury in rats and Sm-M (120-1200 mg/kg, p.o.) lowered dose-dependently the level of lipid peroxidation in liver. PMID- 12748997 TI - Alteration of lethal effects of gamma rays in Swiss albino mice by Tinospora cordifolia. AB - Tinospora cordifolia is widely used in Ayurvedic medicines. It is known for its immunomodulatory, antihepatotoxic, antistress and antioxidant properties. It has been used in combination with other plant products to prepare a number of Ayurvedic preparations. The present study was undertaken to evaluate the radioprotective effect of an aqueous extract of Tinospora cordifolia (TC) against (60)Co gamma radiation. Oral administration of TC 5 mg/kg body wt to Swiss albino mice 1 h and 15 days prior to whole body radiation exposure (8 Gy) produced a significant protection in terms of survival percentage. After oral administration of TC 10 mg/kg body wt/day to mice 7 days prior to whole body irradiation (8 Gy) there was no mortality until day 13 and 50% of the animals survived until day 30. Mice exposed to radiation (8 Gy) without TC pretreatment exhibited signs of radiation sickness such as anorexia, lethargy, ruffled hair, diarrhoea and these animals died within 14 days of irradiation. The results from the present study suggest that Tinospora cordifolia has a radioprotective effect in Swiss albino mice, thereby enhancing the survival of mice against a sublethal dose of gamma radiation. PMID- 12748998 TI - Clinical evaluation of Acalypha ointment in the treatment of superficial fungal skin diseases. AB - In an open non-comparative study to evaluate the efficacy and safety of Acalypha wilkesiana ointment in superficial fungal skin diseases, 32 Nigerian patients with clinical and mycological evidence of superficial mycoses were recruited. Twelve patients defaulted and were lost to follow up, while one patient withdrew because of intolerable excoriation at the site of the lesion. Of the 19 patients that completed the trial, clinical cure was achieved in 73.3% of the patients. The ointment was very efficacious in the treatment of Tinea pedis, Pityriasis versicolor and Candida intetrigo where the cure rate was 100% in each condition. It is recommended that Acalypha ointment can be used for the treatment of these superficial mycoses. PMID- 12748999 TI - Antibacterial efficacy of Rumex nepalensis Spreng. roots. AB - The antibacterial property of Rumex nepalensis Spreng. was evaluated against some strains of bacteria. The methanol extract of the roots (tested at 200-1000 micro g/disc) showed significant concentration-dependent antibacterial activity. PMID- 12749000 TI - Antiviral potency of mistletoe (Viscum album ssp. album) extracts against human parainfluenza virus type 2 in Vero cells. AB - Various extracts from the leaves of mistletoe (Viscum album L. ssp. album) were investigated for their antiviral activity on human parainfluenza virus type 2 (HPIV-2) growth in Vero cells. Plant extracts were prepared using distilled water, 50% ethanol, petroleum ether, chloroform and acetone. The 50% effective dose (ED(50)) of aqueous extract for HPIV-2 replication was 0.53 +/- 0.12 micro g/mL, and the antiviral index (AI), which was based on the ratio of the 50% inhibitory concentration (CD(50)) for host cell viability to the ED(50) for parainfluenza virus replication, was 10.05. The aqueous extract was found to be the most selective inhibitor. Furthermore, the aqueous extract at a concentration of 1 micro g/mL was found to inhibit HPIV-2 replication and the virus production was suppressed to more than 99% without any toxic effect on host cells. The chloroform extract was also found to be moderately active. In an effort to further analyse the mechanism of antiviral activity, the effectiveness of the aqueous extract on different steps of virus replication was examined. The antiviral activity could neither be attributed to the direct inactivation of the HPIV-2 nor to the inhibition of adsorption to Vero cells. The active aqueous extract has shown a dose-dependent antiviral activity on virus replication. PMID- 12749002 TI - Superoxide dismutase activity enhanced by green tea inhibits lipid accumulation in 3T3-L1 cells. AB - We studied the effect of powdered green tea on intracellular superoxide dismutase activity in the adipose conversion of 3T3-L1 cells. By the 14 days of culture with insulin, the triglyceride concentration was increased. When powdered green tea and insulin were added simultaneously, the increased triglyceride content was decreased (p < 0.05), and the superoxide dismutase activities were significantly enhanced (p < 0.05). These data suggest that green tea may have an antilipogenic activity due to its radical scavenging activity mechanism. PMID- 12749001 TI - Hepatoprotective phenolic constituents of Rhodiola sachalinensis on tacrine induced cytotoxicity in Hep G2 cells. AB - Two hepatoprotective phenolic compounds, kaempferol (2) and salidroside (4), were isolated from the roots of Rhodiola sachalinensis together with two inactive compounds cinnamyl alcohol (1) and daucosterol (3) based on the hepatoprotective activity against tacrine-induced cytotoxicity in human liver-derived Hep G2 cells. The EC(50) values of compounds 2 and 4 were 33.5 and 51.3 micro m, respectively. Silybin as a positive control showed an EC(50) value of 68.4 micro m. PMID- 12749003 TI - Inhibitory effect of Adonis amurensis components on tube-like formation of human umbilical venous cells. AB - Antiangiogenic activity-guided fractionation and isolation carried out on the methanol extract of Adonis amurensis led to the identification of three compounds, namely cymarin, cymarol, and cymarilic acid. Amongst the three compounds, cymarilic acid was isolated from this plant for the first time. This compound showed no significant cytotoxicity against tumor cell lines but was found to be strongly inhibitory toward tube formation induced by human umbilical venous endothelial (HUVE) cells. Cymarin and cymarol exhibited potent cytotoxicity against a human solid tumor cell line A549 (human lung carcinoma), while being inactive on murine leukemic cells (L1210). PMID- 12749004 TI - Glycosmis arborea extract as a hepatoprotective agent. AB - Glycosmis arborea is a plant possessing various medicinal properties. The aim of the present study was to investigate the hepatoprotective efficacy of the butanol extract obtained from the aerial parts of the plant. The test sample was prepared by extracting the material through different steps. The extract thus obtained was dissolved in normal saline. Albino rats were prophylactically treated with the extract (i.p.) for 3 weeks. At the end of 3rd week all the groups were injected with hepatotoxic agents. After 48 h of injection, blood was collected and livers were taken out. Different enzymes in the serum were assayed and histopathological study was performed with liver. Glycosmis arborea extract was able to overcome the toxic effects of hepatotoxic agents in terms of lowering the levels of serum GPT, alkaline phosphatase and increased level of SOD in serum. TBARS generation in liver was also altered. Moreover, necrosis of liver produced by carbon tetrachloride was reversed by the extract. PMID- 12749005 TI - Two isoflavones and bioactivity spectrum of the crude extracts of Iris germanica rhizomes. AB - In vitro biological activities including bactericidal, fungicidal and insecticidal activities as well as phytotoxicity and brine shrimp toxicity of the petroleum ether, chloroform and ethyl acetate extracts of Iris germanica L. were determined. The bactericidal activity of the extracts was assayed by the agar well diffusion test. In the fungicidal test, the agar tube dilution method was used. The insecticidal activity was determined by the exposure method. The toxicity of the extracts was evaluated by the phytotoxicity test as well as the brine shrimp toxicity test. The chloroform and ethyl acetate extracts of I. germanica rhizomes exhibited bactericidal activity, while the petroleum ether extract did not exhibit any bactericidal, fungicidal and insecticidal activities. It was also inactive in the brine shrimp toxicity test, whereas it showed significant phytotoxicity against the plant Lemna aequinoctialis Welv. Two known isoflavones were isolated from the chloroform extract of the plant. PMID- 12749007 TI - Biology, clinical, and hematologic features of acute megakaryoblastic leukemia in children. AB - To assess the incidence, clinical features at presentation, hematologic, immunophenotypic, and cytogenetic characteristics of AMKL in children we prospectively studied 834 consecutive non selected children with newly diagnosed acute leukemia (AL) admitted to the Hematology Department at the Instituto Nacional de Pediatria (INP), Mexico, D.F. We found 682 cases (81.8%) with a typical ALL immunophenotype, and the remaining 152 (18.2%) were considered to have AML. In 29 of the 152 patients with AML studied, a diagnosis of AMKL was established. These 29 cases represented 19.1% of the cases of AML and 3.48% of the total cases of AL during the time span covered by the study. Twenty-four percent of the cases occurred in infants 2 years old or younger and 41.4% occurred in children 41 months of age or younger. In contrast, in only 18.6% of the patients with AML (M0-M6), the diagnosis was established before 42 months of age and in 17% before their second year of life. Clinical presentation was not strikingly different than that observed in patients with other types of AML, and the time interval from onset of symptoms to diagnosis was also similar, though in a small subset of patients, the clinical course was characterized by a chronic slowly progressive disorder extending over weeks or months resembling smoldering leukemia or chronic myelofibrosis with agnogenic myeloid metaplasia. Bone marrow (BM) fibrosis was a constant features in our patients; 75% of the patients studied showed this complication at the time of diagnosis. Some rather unusual findings in this study were intense skeletal pains from multiple osteolytic lesions, the presence of soft-tissue tumor, and the presence of cohesive scanty clusters of primitive-looking blast cells in BM aspirates. Several interesting cytogenetic findings in our study were t(1;22)(p13;q13) in a 14-year-old boy, t(9;22)(q34;q11) in one patient, and monosomy 7 in two patients. Another important finding in our study was the clinical association with colonic adenocarcinoma in one patient, an association that to our knowledge has not been reported previously. In conclusion, our data suggest that the incidence of AMKL in Mexico might be higher than those reported in Caucasian white pediatric population, and that biologic and cytogenetic profile may differ from those of western countries, but more studies are needed to corroborate cytogenetic heterogeneity, ethnic and geographic diversity. Early onset of the disease, low WBC counts, slight thrombocytopenia or normal platelet counts, and BM fibrosis were characteristic distinctive features of at least half of the patients with this subtype of AML. PMID- 12749008 TI - Factor V Leiden and prothrombin gene G20210A mutation in children with cerebral thromboembolism. AB - We investigated whether there is an association between factor V Leiden (FVL) and/or prothrombin gene G20210A mutation (PT20210A) and cerebral thromboembolism in a pediatric Argentinean population. From May 1992 to January 2002, 44 consecutive children with arterial ischemic stroke (AIS) and 23 children with cerebral sinovenous thrombosis (SVT) were prospectively studied at a single center. The prevalence of both mutations was compared with a 102 age-matched controls. In children with AIS, the frequencies (patients vs. controls), odds ratio (OR), and 95% confidence interval (95% CI) for the presence of FVL were as follows: 2.3% vs. 2%, OR/95% CI, 1.16/0.2 to 13.2; P value = 0.99. No cases of PT20210A were found in this group. In children with SVT, the frequencies (patients vs. controls), OR, and 95% CI were as follows: FVL (4.3% vs. 2%, OR/95% CI, 2.27/0.22 to 6.2; P value = 0.99) and PT20210A (4.3% vs. 1%; OR/95% CI, 4.6/0.3 to 76.3; P value = 0.3354). One child with PT20210A also had an inherited protein C deficiency. In 12 (18%) out of the 67 children with cerebral thromboembolism, without the aforementioned mutations, other prothrombotic disorders were detected. Although a multi-center prospective study with a large number of Argentinean pediatric patients is needed to obtain considerable evidence, no association between factor V Leiden and/or prothrombin gene G20210A mutation and cerebral thromboembolism was found in this pediatric series. PMID- 12749010 TI - Intravenous immunoglobulin therapy results in post-infusional hyperproteinemia, increased serum viscosity, and pseudohyponatremia. AB - Intravenous immunoglobulin (IVIG) therapy is associated with rare reports of thromboembolic events and severe hyponatremia. We hypothesized that IVIG therapy may result in hyperproteinemia, increased serum viscosity, and pseudohyponatremia. We conducted a prospective observational study to evaluate the incidence of hyperproteinemia occurring after IVIG therapy and its relationship to serum sodium, viscosity, osmolality, and the serum osmolar gap. Eighteen IVIG infusions at a standard dose of 2 g/kg administered over 2-5 days were evaluated. Serum glucose, sodium, protein, viscosity, osmolality, and a calculated osmolar gap were obtained prior to therapy, 6 hr after the initiation of therapy, 24 hr after the conclusion of therapy, and on post-treatment day 10. Paired t-testing revealed a statistically significant increase in serum protein and viscosity and decrease in serum sodium and calculated osmolality 24 hr after the completion of IVIG therapy. The calculated serum osmolar gap increased insignificantly. In multivariate analysis, hyperproteinemia at the 6-hr time point predicted hyponatremia (P < 0.000), and hyperproteinemia at the 24-hr time point predicted both hyponatremia and increased serum viscosity (P = 0.024). These data demonstrate that increased serum viscosity occurs following IVIG therapy due to hyperproteinemia, and the rare hyponatremia reported is a pseudohyponatremia also due to hyperproteinemia. PMID- 12749009 TI - Clinicopathologic analysis of ocular adnexal lymphomas: extranodal marginal zone b-cell lymphoma constitutes the vast majority of ocular lymphomas among Koreans and affects younger patients. AB - The majority of ocular adnexal lymphomas (OAL) are primary marginal zone B-cell lymphomas (MALT lymphomas). The present study correlated the clinicopathological variables with the histologic subtypes by World Health Organization (WHO) classification with emphasis on MALT lymphomas in OALs of Koreans. There were 68 cases (31 males and 37 females), with a mean age of 45.9 years (range 7-89 years). Histologically, 61 MALT-type, 2 diffuse large B-cell (DLBCL), 2 mantle cell type (MCL), 1 anaplastic large-cell (ALCL), and 2 NK/T-cell lymphomas (NK/T L) were counted among them. Fifty-seven were primary cases (P-OAL), and 11 were secondary cases (S-OAL). Nearly all P-OALs were MALT lymphomas (n = 56, 98%), with an exception of 1 MCL. Eleven S-OALs included 5 MALT type, 2 DLBCL, 1 ALCL, 1 MCL, and 2 NK/T-L. All MALT lymphoma patients were alive (n = 59) except for 2 after a mean duration of follow-up of 27.6 months (range: 0-108 months): one died of an unrelated cause and one died of recurrence. One non-MALT type P-OAL was alive with no evidence of disease (42 months). Of the 11 S-OAL, 4 had marrow involvement and 5 had progression or relapse outside the orbit. Compared with the other subtypes, MALT lymphoma was more likely to present with local disease (P = 0.001), achieve complete remission (CR) (0.022), and be alive at last follow-up (0.197), and less likely to experience recurrence (P = 0.06). In conclusion, OALs in Koreans are characterized by a preponderance of primary lymphomas over systemic lymphomas, striking predominance of MALT type lymphomas, and young age of occurrence. Histologic subtype by WHO classification has a significant correlation with the final outcome, with the most favorable outcome associated with OALs of the MALT type. PMID- 12749011 TI - Prognostic significance of bcl-2, bax, and p53 expression in diffuse large B-cell lymphoma. AB - Diffuse large B-cell lymphoma (DLCL) exhibits heterogeneous clinical features and varies markedly in response to treatment and prognosis. Because apoptosis-related proteins may play an important role in predicting the prognosis of DLCL, the current study investigated the prognostic significance of a high level of bcl-2, bax, and p53 expression in relation to clinical characteristics in patients with DLCL. Paraffin-embedded specimens from 94 patients with de novo DLCL were analyzed immunohistochemically for bcl-2, bax, and p53 gene expression. Cases with a positive immunohistological stain in more than 50% of the tumor cells were considered to have DLCL-positive expression. Patients were treated optimally, i.e., with radiotherapy including brief cycles of CHOP or CHOP-like regimens for patients with stage 1-2A diseases and with at least 6 cycles of CHOP or CHOP-like regimens for stage 2B-4 diseases. The responses to therapy and survival were then analyzed in 94 uniformly staged patients. bcl-2 expression was identified in 24 patients (26.4%), bax expression in 35 patients (37.6 %), and p53 expression in 21 patients (22.6%). bax expression proved to be a statistically significant prognostic factor in predicting the overall survival (OS) (P = 0.0015) and disease-free survival (DFS) (P = 0.0052), regardless of other clinical factors or immunohistological results. There was no significant difference in the OS (P = 0.0682) or DFS (P = 0.088) between the bcl-2-positive (n = 24) and bcl-2-negative (n = 67) groups. However, bcl-2 expression was found to be unfavorably associated with the OS (P = 0.0054) in a confined group with low (n = 51) or low intermediate (n = 22) IPI scores. The expression of p53 exhibited no statistical correlation with the OS or DFS. A multivariate analysis revealed that IPI score, bulky mass, and bax expression were all significantly associated with the DFS or OS. bax and bcl-2 should be considered as independent biologic prognostic parameters in DLCL, thereby aiding in the identification of patient risk groups. As such, bcl-2-positive patients with a low or low intermediate IPI score, or without a high level of bax expression could be candidates for more intensive therapy or alternative therapeutic approaches. PMID- 12749012 TI - Health status and health-related quality of life associated with von Willebrand disease. AB - Von Willebrand disease (VWD) is the commonest inherited disorder of hemostasis and the majority of women with this disorder experience excessive uterine bleeding. Yet very little information is available on the health-related quality of life (HRQL) in individuals with VWD. To test the a priori hypotheses that these individuals will have poorer HRQL than members of the general population, and that this burden of morbidity will correlate with the severity of VWD, a cross-sectional study was undertaken of a population-based cohort in a regional hemophilia program in Ontario, Canada. A survey was made of individuals over 13 years of age with VWD who self-reported their health status using a standard 15 item questionnaire. The responses were converted to levels in the Health Utilities Index Mark 2 (HUI2) and Mark 3 (HUI3) health status classification systems to form multi-element vectors from which single attribute morbidity and overall HRQL utility scores were determined. As a group, individuals with VWD were shown to have poorer HRQL than members of the general population and those with Type 2 disease carried a greater burden of overall morbidity than those with Type 1 disorder. Morbidity was evident mainly in the attributes of emotion, cognition with pain. A striking difference was observed between males and females, with the latter having overall HRQL utility scores similar to those reported previously for HIV positive, severe hemophiliacs. It is possible that this remarkable burden of morbidity reflects chronic iron deficiency associated with menorrhagia. A national study has been proposed to address this likelihood as it offers an opportunity for effective therapeutic intervention (iron supplementation) with a concomitant gain in health status and HRQL. PMID- 12749013 TI - Chemotaxis of non-compressed blood polymorphonuclear leukocytes from an adolescent with severe leukocyte adhesion deficiency. AB - We have defined the defect in a child with severe leukocyte adhesion deficiency-1 (LAD) as resulting from a single amino acid shift in CD18 (from a C to T mutation at position 533) that prevents heterodimerization with the CD11 antigens to produce beta(2) integrins-the first reported patient homozygous for this defect. Although beset by frequent infections, the patient has survived to adolescence despite the lack of these important adhesion molecules. Consistent with his clinical course is the ability of his PMN to respond chemotactically in slide preparations, albeit with difficulty because of their poor purchase on substrate. The operant adhesins are unknown; his polymorphonuclear leukocytes (PMN) remain chemotactically responsive in the presence of antibodies to alphavbeta(3) and beta(1) integrins and to integrin-associated protein (IAP). These findings indicate that not all patients with severe LAD are candidates for early bone marrow transplantation. PMID- 12749014 TI - Role of phlebotomy in the management of hemoglobin SC disease: case report and review of the literature. AB - Marked variability is a keynote in the disease course of patients with hemoglobin SC (Hb SC) and hemoglobin S/beta(+)-thalassemia (Hb S/beta(+)-thal), with some patients having a frequency of complications and painful episodes similar to patients with homozygous sickle cell (Hb SS) disease. One possible explanation is that the higher hematocrit in these syndromes may contribute to an increase in blood viscosity, leading to vaso-occlusive pain episodes as well as an increased incidence of thromboembolic complications and retinopathy. We present a patient with Hb SC disease with an excellent baseline functional status who developed splenic infarction at a high altitude. Following splenectomy, the patient developed a sustained increase in hematocrit, an increase in the frequency of painful episodes, as well as new-onset dizziness and malaise. We initiated a therapeutic phlebotomy program in order to lower the hematocrit to pre splenectomy values, as well as to induce iron deficiency. Repeated phlebotomy resulted in a dramatic decrease in symptoms. Our patient no longer requires narcotic analgesics for pain, has resolution of constitutional symptoms, and has not required further hospitalizations for vaso-occlusive pain crises. The correlation between symptoms and hematocrit levels supports the importance of blood viscosity in contributing to this patient's symptoms. A trial of phlebotomy to reduce viscosity in patients with higher hematocrit values should be considered as an intervention for symptomatic patients with sickle cell disease. PMID- 12749015 TI - Natural killer cell-type body cavity lymphoma following chronic active Epstein Barr virus infection. AB - We describe a 69-year-old female who developed natural killer cell-type body cavity lymphoma following chronic active Epstein-Barr virus (CAEBV) infection. Examination of the patient's pleural effusion revealed large abnormal lymphocytes, which were CD2(+), CD7(+), CD30(+), CD56(+), CD3(-), and CD4(-). No rearrangement of T cell receptor genes was detected. Clonal proliferation of Epstein-Barr virus (EBV)-infected cells in pleural effusion was demonstrated by Southern blot hybridization analysis. Human herpesvirus type-8 (HHV-8) DNA was not detected in these cells. The patient achieved a complete remission with combination chemotherapy. Prior to the clinical onset of lymphoma, high fever of unknown origin had persisted for 21 months. IgG antibodies to EBV-viral capsid antigen and to EBV-early antigens, types D and R were not high (1:160 and less than 1:10, respectively). Two months after the onset of fever, however, retrospective quantitative PCR assay revealed a high EBV DNA load in plasma, indicating that CAEBV infection had been the cause of the patient's recurrent fever. The remarkable features of this case are (i) the development of lymphoma following CAEBV infection that demonstrated a normal pattern of EBV-specific antibodies, (ii) the development of HHV-8-negative body cavity lymphoma, and (iii) the effectiveness of combination chemotherapy. PMID- 12749016 TI - Pulmonary and intracerebral plasmacytomas in a patient without multiple myeloma: a case report. AB - Extramedullary plasmacytoma of the lung or brain parenchyma is rare. We report herein a case of extramedullary plasmacytoma involving both the lung and parenchymal brain. We review the literature, comparing and contrasting the current case. PMID- 12749017 TI - Complete recovery from refractory immune thrombocytopenic purpura in three patients treated with etanercept. AB - Management of patients with immune thrombocytopenic purpura (ITP) who have persistent, severe, and symptomatic thrombocytopenia following splenectomy is difficult and empirical. No single agent or regimen provides long-term success for most patients, and for most treatments it is difficult to assess whether benefits outweigh risks. We report three consecutive patients with critical chronic refractory ITP, who responded promptly and completely following treatment with etanercept, an inhibitor of tumor necrosis factor-alpha. These patients had failed 6-11 previous treatments. In the first patient, etanercept was given for its approved indication: a flare of co-existing rheumatoid arthritis. The next two patients were treated with etanercept because of successful outcomes in the previous patients. Although etanercept appeared to be effective treatment for ITP in these 3 patients, the experimental nature of this treatment and the potential risks must be emphasized. On the basis of these case reports, a clinical trial has been initiated to systematically evaluate the efficacy and risks of etanercept in the management of children and adults with chronic ITP. PMID- 12749018 TI - Charcot Leyden crystals in acute myeloid leukemia. PMID- 12749019 TI - Changes in subcellular distribution of protocadherin gamma proteins accompany maturation of spinal neurons. AB - Protocadherins gamma (Pcdhgamma) are a family of transmembrane proteins in which variable extracellular domains are associated with an invariant cytoplasmic domain, potentially allowing these proteins to trigger common cellular responses through diverse extracellular signals. We studied the expression of the family by in situ hybridisation and immunohistochemistry for the conserved portion of the mRNA or protein. During mouse development, Pcdhgamma expression is highest in neural tissues, but is also present in some nonneural tissues. In the adult, Pcdhgamma expression is maintained at high levels in brain, in particular in hippocampus and in the Purkinje cells of the cerebellum, whereas it is downregulated in spinal cord. Using antibodies against the conserved cytoplasmic domain, we show that in cultured embryonic spinal cord neurons, Pcdhgamma protein is present initially in both axonal and dendritic growth cones. At later stages of differentiation in vitro, Pcdhgamma distribution becomes polarised to the somatodendritic compartment. We propose that members of the Pcdhgamma family may play roles in neuronal growth and maturation. PMID- 12749020 TI - EGF and NGF injected into the brain of old mice enhance BDNF and ChAT in proliferating subventricular zone. AB - The response of cells localized in the brain subventricular zone (SVZ) to growth factor stimulation has been largely described for development and adult life, whereas no information on their behavior during aging is available. To address the question of whether the cells in the SVZ of old mice respond to the intracerebroventricular administration of epidermal growth factor (EGF) and nerve growth factor (NGF), we studied the distribution of proliferating cells and the effects on ChAT and brain-derived neurotrophic factor (BDNF) synthesis in forebrain and SVZ. It was found that the conjoint administration of EGF + NGF produced a major increase in ChAT expression in both forebrain and SVZ. The ChAT mRNA levels and the number of ChAT positive cells localized in the ventricular border and in the parenchyma of SVZ area were also increased significantly in the mice receiving EGF + NGF. Enhanced numbers of SVZ cells expressing proliferative markers were also discovered in EGF + NGF treated mice and some of these cells expressed cholinergic markers, as demonstrated by double immunostaining. In addition, EGF and NGF treatments significantly upregulate BDNF protein and mRNA levels in this brain region. The present study demonstrates that cells localized in SVZ of aged mouse brain retain the capacity to respond to EGF and NGF and that after stimulation with these two growth factors, the synthesis of ChAT and BDNF also increases. The implication that cells of the SVZ remain a reservoir of cholinergic and BDNF-positive neurons in aged brain opens a new perspective for understanding the role of growth factors during neurodegenerative disorders associated with aging. PMID- 12749021 TI - Rapid method for culturing embryonic neuron-glial cell cocultures. AB - A streamlined, simple technique for primary cell culture from E17 rat tissue is presented. In an attempt to standardize culturing methods for all neuronal cell types in the embryo, we evaluated a commercial medium without serum and used similar times for trypsinization and tested different surfaces for plating. In 1 day, using one method and a single medium, it is possible to produce robust E17 cultures of dorsal root ganglia (DRG), cerebellum, and enteric plexi. Allowing the endogenous glial cells to repopulate the cultures saves time compared with existing techniques, in which glial cells are added to cultures first treated with antimitotic agents. It also ensures that all the cells present in vivo will be present in the culture. Myelination commences after approximately 2 weeks in culture for dissociated DRG and 3-4 weeks in cerebellar cultures. In enteric cultures, glial wrapping of the enteric neurons is seen after 3 weeks (2 weeks in ascorbic acid), suggesting that basal lamina production is important even for glial ensheathment in the enteric nervous system. No overgrowth of fibroblasts or other nonneuronal cells was noted in any cultures, and myelination of the peripheral nervous system and central nervous system cultures was very robust. PMID- 12749022 TI - Treatment with metallothionein prevents demyelination and axonal damage and increases oligodendrocyte precursors and tissue repair during experimental autoimmune encephalomyelitis. AB - Experimental autoimmune encephalomyelitis (EAE) is an animal model for the human demyelinating disease multiple sclerosis (MS). EAE and MS are characterized by significant inflammation, demyelination, neuroglial damage, and cell death. Metallothionein-I and -II (MT-I + II) are antiinflammatory and neuroprotective proteins that are expressed during EAE and MS. We have shown recently that exogenous administration of Zn-MT-II to Lewis rats with EAE significantly reduced clinical symptoms and the inflammatory response, oxidative stress, and apoptosis of the infiltrated central nervous system areas. We show for the first time that Zn-MT-II treatment during EAE significantly prevents demyelination and axonal damage and transection, and stimulates oligodendroglial regeneration from precursor cells, as well as the expression of the growth factors basic fibroblast growth factor (bFGF), transforming growth factor (TGF)beta, neurotrophin-3 (NT 3), NT-4/5, and nerve growth factor (NGF). These beneficial effects of Zn-MT-II treatment could not be attributable to its zinc content per se. The present results support further the use of Zn-MT-II as a safe and successful therapy for multiple sclerosis. PMID- 12749024 TI - Vasoactive intestinal peptide and nitric oxide promote survival of adult rat myenteric neurons in culture. AB - Several motility disorders originate in the enteric nervous system (ENS). Our knowledge of factors governing survival of the ENS is poor. Changes in the expression of vasoactive intestinal peptide (VIP) and nitric oxide synthase (NOS) in enteric neurons occur after neuronal injury and in intestinal adaptation. The aim of this study was to evaluate whether VIP and nitric oxide (NO) influence survival of cultured, dissociated myenteric neurons. Neuronal survival was evaluated after 0, 4, and 8 days in culture. Influence of VIP and NO on neuronal survival was examined after culturing in the presence of VIP, NO donor, VIP antiserum, or NOS inhibitor. A marked loss of neurons was noted during culturing. VIP and NO significantly promoted neuronal survival. Corroborating this was the finding of an enhanced neuronal cell loss when cultures were grown in the presence of VIP antiserum or NOS inhibitor. PMID- 12749023 TI - Myelination and motor coordination are increased in transferrin transgenic mice. AB - Myelin deficiency in the central nervous system (CNS) can cause severe disabling conditions. Most of the transgenic mice models overexpressing myelin components have limitations for investigators of myelin deficiency and myelin therapy as they severely alter CNS architecture. It has been postulated that transferrin (Tf) is involved in oligodendrocyte (OL) maturation and myelinogenesis. Because Tf is not an intrinsic myelin constituent, we decided to investigate if its overexpression could have an impact on the myelination process without affecting myelin integrity. We generated transgenic mice containing the complete human Tf gene specifically overexpressed in OLs. This overexpression leads to more than a 30% increase in myelin components, such as galactolipids, phospholipids, and proteins. Electron microscopy showed that myelin is structurally normal in terms of thickness and compaction. Behavior analysis showed that mice do not display significant modifications in their locomotion and cognitive and emotional abilities. Furthermore, in one of the genetic background, animals presented a significant increase in motor coordination. We did not find any modification in OL number during early postnatal development, suggesting that Tf does not act on OL proliferation. In addition, the levels of iron and ferritin remained unchanged in the brain of transgenic mice compared to control mice. Our findings indicate that, besides its known iron transport function, Tf is able to influence myelination process and induce behavioral improvements in mice. PMID- 12749025 TI - Glucagon-like peptide-1 decreases endogenous amyloid-beta peptide (Abeta) levels and protects hippocampal neurons from death induced by Abeta and iron. AB - Glucagon-like peptide-1(7-36)-amide (GLP-1) is an endogenous insulinotropic peptide that is secreted from the gastrointestinal tract in response to food. It enhances pancreatic islet beta-cell proliferation and glucose-dependent insulin secretion and lowers blood glucose and food intake in patients with type 2 diabetes mellitus. GLP-1 receptors, which are coupled to the cyclic AMP second messenger pathway, are expressed throughout the brains of rodents and humans. It was recently reported that GLP-1 and exendin-4, a naturally occurring, more stable analogue of GLP-1 that binds at the GLP-1 receptor, possess neurotrophic properties and can protect neurons against glutamate-induced apoptosis. We report here that GLP-1 can reduce the levels of amyloid-beta peptide (Abeta) in the brain in vivo and can reduce levels of amyloid precursor protein (APP) in cultured neuronal cells. Moreover, GLP-1 and exendin-4 protect cultured hippocampal neurons against death induced by Abeta and iron, an oxidative insult. Collectively, these data suggest that GLP-1 can modify APP processing and protect against oxidative injury, two actions that suggest a novel therapeutic target for intervention in Alzheimer's disease. PMID- 12749026 TI - Effects of deep hypothermia on nitric oxide-induced cytotoxicity in primary cultures of cortical neurons. AB - Nitric oxide (NO) is thought to play a major role during cerebral ischemia. However, the protective efficacy of hypothermia against NO-induced neurotoxicity remains to be examined. In the present study, the degree of neurotoxicity induced by NO was analyzed in two temperature groups (normothermia, 37 degrees C; deep hypothermia, 22 degrees C) of cultured E16 Wistar rat cortical neurons. Two different NO donors, 1-hydroxy-2-oxo-3-(N-ethyl-2-aminoethyl)-3-ethyl-1-triazene (NOC-12) and 1-hydroxy-2-oxo-3-(3-amynopropyl)-3-isopropyl-1-triazene (NOC-5), that have equal half-lives at 37 degrees C and 22 degrees C, respectively, were used. Cultured neurons in each temperature group were exposed to 30 and 100 micro M NOC for three different time courses, 6 hr, 12 hr, and 24 hr. The survival rates of neurons were evaluated by assessing viable neurons on photomicrographs before and after the experiments. The highest survival rate (approximately 93%) was seen in both temperature groups when neurons were exposed to 30 micro M NOC for 6 hr and 12 hr, and there was no significant difference observed between these two groups (P > 0.05). Almost equal survival rates were observed in both temperature groups following exposure to 30 micro M NOC for 24 hr (at 37 degrees C, 80.4% +/- 2.6%; at 22 degrees C, 83.2% +/- 1.6%; P > 0.05). During exposure to 100 micro M NOC, although the survival rate linearly decreased (approximately from 70% to 5%) in both temperature groups when exposed for 6-24 hr, there were no significant intergroup differences observed (P > 0.05). In conclusion, hypothermia does not provide adequate protection to the neurons by acting on the mechanisms evoked by NO, so we speculate that hypothermia may not confer neuroprotetcion once NO is released during ischemia. PMID- 12749027 TI - Effect of antipsychotic drugs on brain-derived neurotrophic factor expression under reduced N-methyl-D-aspartate receptor activity. AB - Brain-derived neurotrophic factor (BDNF) promotes a variety of neuromodulatory processes during development as well as in adulthood. This neurotrophin has been associated with synaptic plasticity, suggesting that its regulation may represent one of the mechanisms through which psychotropic drugs alter brain function. Because reduced glutamatergic function represents a major feature of schizophrenia, we investigated the effects of the concomitant administration of haloperidol or olanzapine with the N-methyl-D-aspartate (NMDA) receptor antagonist MK-801 on BDNF expression. MK-801 reduces the hippocampal expression of the neurotrophin; this effect was exacerbated by haloperidol, but it was normalized by olanzapine. Our data reveal a fine tuning of BDNF biosynthesis and a differential modulation by antipsychotic drugs when NMDA-mediated transmission is reduced, suggesting that haloperidol and olanzapine can produce different effects on brain plasticity through the modulation of BDNF expression. PMID- 12749028 TI - Light exposure during daytime modulates expression of Per1 and Per2 clock genes in the suprachiasmatic nuclei of mice. AB - The suprachiasmatic nuclei (SCN) of the hypothalamus contain the master circadian clock in mammals. Nocturnal light pulses that reset the circadian clock also lead to rapid increases in levels of Per1 and Per2 mRNA in the SCN, suggesting that these genes are involved in the synchronization to light. During the day, when light has no phase-shifting effects in nocturnal rodents, the consequences of light exposure for Per expression have been less thoroughly studied. Therefore, the effects of light exposure during the day were assessed on Per1 and Per2 mRNA in the SCN of mice. Expression of Per1 and Per2 was generally increased by 30-min light pulses during the subjective day, with more pronounced effects in the morning. One exception was noted for a transient decrease in Per2 expression after a short light pulse applied at midday. Prolonged light exposure (up to 3 hr) starting at midday markedly increased Per2 expression but not that of Per1. Moreover, the amplitude of the daily variations of both Per and the duration of Per1 peak was increased in mice exposed to a light-dark cycle compared with those transferred to constant darkness. Finally, the amplitude of the daily variations of both Per and the basal level of Per1 were increased in mice under a light-dark cycle compared with animals synchronized to a skeleton photoperiod (i.e., with daily dawn and dusk 1-hr exposures to light). Taken together, the results indicate that prolonged light exposure during daytime positively modulates daily levels of Per1 and Per2 mRNA in the SCN of mice. PMID- 12749029 TI - Altered CTX-catalyzed and endogenous [32P]ADP-ribosylation of stimulatory G protein alphas isoforms in postmortem bipolar affective disorder temporal cortex. AB - Reports of elevated Gs alpha subunit (alpha(s)) immunolabeling and cAMP-mediated hyper-functionality in autopsied cerebral cortical brain regions from bipolar affective disorder (BD) patients suggest signal transduction abnormalities occur in this disorder. Because covalent modification of alpha(s) can affect its turnover and levels, we determined whether CTX-catalyzed and endogenous [(32)P] adenosine diphosphate (ADP)-ribosylation of alpha(s) isoforms are altered in temporal and occipital cortical regions, which show elevated alpha(s) levels in BD as compared to nonpsychiatric subjects. Reduced CTX-catalyzed [(32)P]ADP ribosylated alpha(s-S) and endogenous [(32)P]ADP-ribosylation of a 39-kDa alpha(s)-like protein were found in BD temporal cortex compared to controls. These findings suggest that clearance of these alpha(s) isoforms through ADP ribosylation may be decreased in BD temporal cortex. Although no differences were observed in mean levels of endogenous and CTX-catalyzed [(32)P]ADP-ribosylation of alpha(s-L) in BD temporal cortex, alpha(s-L) immunolabeling was elevated significantly and correlated inversely with the degree of endogenous [(32)P]ADP ribosylation of this subunit. In addition, endogenous [(32)P]ADP-ribosylation of an exogenous substrate, myelin basic protein, was similar in BD and comparison subject temporal cortex. Taken together, these observations suggest that elevations of alpha(s) in BD brain are more likely related to factors affecting the disposition or availability of alpha(s) to this posttranslational enzymatic modification. PMID- 12749030 TI - Tendon chitosan tubes covalently coupled with synthesized laminin peptides facilitate nerve regeneration in vivo. AB - We have developed tendon chitosan tubes having the ability to bind peptides covalently, and the effectiveness of laminin peptides coupled to these tubular wall on nerve regeneration was examined in vivo. Bridge graft implantation (15 mm) into the sciatic nerve of SD rats was carried out using chitosan tubes having a triangular cross section containing either covalently bound intact laminin or the laminin peptides CDPGYIGSR or CSRARKQAASIKVAVSAD or being nontreated (N = 20 in each group). As a control, isografting (N = 5) was carried out. Three rats in each experimental group were sacrificed for histology observations after 1, 2, 4, 6, and 8 weeks. The total area of regenerating tissue in the tube and the length of the area where regenerating tissue attached to the inner surface of the tube were measured. In five rats from each experimental and control group, the latency quotient between the implanted and the nontreated site was determined 12 weeks after implantation. Furthermore, the percentage of myelinated axon area was measured at a 10-mm distance from the distal anastomosed site. Histological findings suggest that the immobilized laminin, confirmed by immunostaining as long as 12 weeks postoperatively, as well as laminin oligopeptides may effectively assist nerve tissue extension. According to statistical analysis of the percentage neural tissue found in relation to evoked action potentials, the sequential treatments with YIGSR first followed by IKVAV matched the effectiveness of intact laminin in enhancing nerve regeneration. However, when compared with that after isografting, the enhancement of regenerated axon growth was less sufficient. PMID- 12749031 TI - Cell-free fetal DNA concentration in plasma of patients with abnormal uterine artery Doppler waveform and intrauterine growth restriction--a pilot study. AB - OBJECTIVE: To evaluate if an increased amount of fetal DNA concentration can be found in women screened positive for intrauterine growth restriction because of abnormal uterine artery Doppler waveforms. METHODS: We enrolled eight pregnant women (each bearing a male fetus), with the evidence of abnormal uterine artery Doppler waveforms, and 16 control patients for a case-control study matched for gestational age (1 : 2). Uterine artery Doppler was carried out at 20 to 35 weeks' gestation (median 29). The mean uterine artery resistance index (RI) was subsequently calculated, and a value >0.6 was considered positive for the clinical features of pre-eclampsia. The SRY locus was used to determine the amount of male fetal DNA in the maternal plasma at the time of Doppler analysis. RESULTS: Two controls (normal Doppler) were excluded from the final analysis because they had a pre-term delivery. One case (abnormal Doppler) had evidence of intrauterine growth restriction at the time of enrolment. In four out of eight cases (abnormal Doppler), intrauterine growth restriction was subsequently observed. Multiples of median (MoM) conversion of the fetal DNA values showed an increase of 1.81 times in the cases when compared to the controls. An increase of 2.16 times was instead observed for the cases with a growth-restricted fetus (5 cases out of 8) in comparison with the controls (14 cases). CONCLUSIONS: In subjects positive to uterine artery Doppler velocimetry analysis (Doppler analysis for pre-eclampsia screening), the fetal DNA concentration is higher than expected, in the absence of any other clinical feature. Since the increase in fetal DNA seems to be related to the presence or to the future development of intrauterine growth restriction, this paper suggests a possible integration between ultrasound and molecular markers for predicting the disease in some cases. PMID- 12749032 TI - A new report of mesomelic camptomelia, polydactyly and Dandy-Walker complex in siblings. AB - Two male siblings with several malformations are reported. The anomalies detected in both fetuses were mesomelic camptomelia, postaxial hexadactyly and Dandy Walker complex. There was only one similar previous report in the literature. This combination could represent a specific pattern of malformation or a new syndrome, with different variants. The parents' consanguinity and the recurrence in a subsequent pregnancy suggest an autosomal recessive inheritance pattern. PMID- 12749033 TI - Prenatal diagnosis of de novo terminal deletion of chromosome 7q. AB - OBJECTIVES: To present the prenatal diagnosis and perinatal findings of a de novo terminal deletion of chromosome 7q. CASE: Amniocentesis was performed at 21-weeks gestation owing to a positive result of maternal serum multiple-marker screening. The 30-year-old woman, gravida 2, para 1, had a maternal serum multiple-marker screening test at 18-weeks gestation. The risk of Down syndrome was 1/11 calculated from the gestational age, maternal age, a maternal serum alpha fetoprotein level of 1.026 multiples of the median (MOM), and a maternal serum free beta-human chorionic gonadotrophin (hCG) level of 8.678 MoM. Cytogenetic analysis of the cultured amniotic fluid cells revealed a de novo terminal deletion of 7q, 46,XX,del(7)(q35). Ultrasonography showed intrauterine growth restriction, microcephaly, and tetralogy of Fallot. The pregnancy was terminated subsequently. Grossly, the placenta was normal. On autopsy, the proband additionally manifested a prominent forehead, hypertelorism, epicanthus, upslanting palpebral fissures, a flat and broad nasal bridge, micrognathia, large low-set ears, overriding toes, and a normal brain. Radiography demonstrated a normal spine. Fluorescence in situ hybridization analysis demonstrated a 7q terminal deletion. Genetic marker analysis showed a maternally derived terminal deletion of chromosome 7(q35-qter). CONCLUSION: Fetuses with a de novo 7q terminal deletion may be associated with a markedly elevated maternal serum hCG level and abnormal sonographic findings of intrauterine growth restriction, microcephaly, and congenital heart defects in the second trimester. PMID- 12749034 TI - Sub-lethal hydrops as a manifestation of dehydrated hereditary stomatocytosis in two consecutive pregnancies. AB - Dehydrated hereditary stomatocytosis (DHS) is a rare congenital hemolytic anemia mapping to 16q23-q24. We showed recently that it is part of a pleiotropic syndrome likely to display pseudohyperkalemia and/or different forms of fetal and placental fluid collections. Here, we report a woman with DHS. She had two consecutive pregnancies associated with severe fetal hydrops. Hydrops would probably have been lethal in the absence of appropriate removal of ascites and excess amniotic fluid. In utero exchange transfusion, performed once, was useless, because anemia was not pronounced enough to be the cause of the hydrops. In both newborns, ascites resolved within a week following birth and never recurred. The association of hydrops and hemolytic anemia suggests the possibility of DHS. Symptomatic treatment of the hydrops assists survival until spontaneous resorption occurs. PMID- 12749035 TI - Frequency and clinical consequences of extremely high maternal serum PAPP-A levels. AB - A multicentre study was carried out to determine the frequency and clinical consequences of extremely high maternal serum pregnancy-associated plasma protein (PAPP)-A. There was a total of 79 pregnancies with PAPP-A exceeding 5.0 multiples of the gestation-specific median in a series of 46 776 pregnancies tested (0.2%) at the 7 collaborating centres. Five pregnancies were lost to follow-up, one miscarried and one with Noonan's syndrome was terminated. Of the remaining 72 that ended in a live birth, one infant had gastroschisis and five pregnancies had obstetric complications: pre-eclampsia, pregnancy-induced hypertension, gestational diabetes and two with growth retardation. Among women with high PAPP A and no complications or adverse outcomes, there was no evidence of a substantial change in the levels of other Down syndrome markers or the extent of nuchal translucency. Three analytical methods were used to assay PAPP-A and yielded different frequencies of extremely high levels (0.05%, 0.4% and 0.6%) possibly owing to cross-reaction with another substance. We conclude that women with high PAPP-A can be reassured that there is no reason to suppose that the outcome of pregnancy will differ from those with normal levels, provided other markers are normal. If, as more centres move their Down syndrome screening practice to the first trimester, additional cases emerge with Noonan's syndrome or gastroschisis and raised PAPP-A, this advice will need to be modified. PMID- 12749036 TI - Centre-specific ultrasound nuchal translucency medians needed for Down syndrome screening. AB - Nuchal translucency (NT) measurements were compared between 13 centres participating in a multi-marker Down syndrome screening program. Results from 4765 women were analysed, and there were highly statistically significant between centre differences after allowing for gestation (P < 0.0001). Examination of maternal serum marker levels, expressed in multiples of the median (MoM) for gestation, showed that this was not due to gestational errors. Regression analysis was carried out to derive an equation with a centre-specific component that could be used to express NT in MoMs. Use of this equation reduced the variance of logNT by 15% compared to a published equation. The equation can be readily modified for use in other centres. PMID- 12749037 TI - Prenatal detection of fetal hemoglobin E gene from maternal plasma. AB - In order to provide a noninvasive prenatal diagnosis of the hemoglobin E (Hb E) related disorder, we have evaluated the possibility of identifying the fetal beta(E)-globin gene in maternal plasma. The analysis was performed during 8 to 18 weeks of gestation using DNA extracted from 200 micro L of plasma from pregnant women whose husbands carried Hb E. The beta(E)-globin mutation in maternal plasma was detected by a nested PCR amplification followed by the Mnl I restriction analysis. The result was compared with that of routine analysis of the CVS specimens. Among the five pregnant women examined, the fetal beta(E)-globin gene was identified in maternal plasma in three of them and the result was completely concordant with the conventional CVS analysis. This simple noninvasive prenatal detection of the fetal beta(E)-globin gene should prove useful in a prevention and control program of Hb E/beta-thalassemia in countries where the beta(E) globin gene is prevalent. PMID- 12749038 TI - Morphological and cytogenetic analysis of intact oocytes and blocked zygotes. AB - We examined cytological and cytogenetic parameters of 1076 oocytes and 385 zygotes that failed to develop post in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). Out of 1076 oocytes, 894 (83%) arrested oocytes showed a first polar body and were thus assumed arrested at metaphase II while the remainder showed no polar body. In the group of oocytes with a polar body, 20.5% had an abnormal karyotype. Cytologically, premature sperm chromosome condensation was noted in 28.3% of uncleaved oocytes. This high PCC can be explained by the different grades of oocyte maturity from one center to another. Oocytes from older women showed no increased aneuploidy but did show increased premature chromosome condensation. Analysis by classical technique of 220 uncleaved zygotes showed 91 with highly condensed chromosomes, 53 with asynchrony of condensation, 31 with pulverized chromosomes, and 45 arrested at the first somatic metaphase. Out of 385 arrested zygotes, 165 were explored by in situ hybridization. FISH using a set of 7 chromosome-specific probes showed aneuploidy in the chromosomes analyzed (13, 16, 18, 21, 22, X, Y) in 21.8% of blocked zygotes (19-25% depending on morphology). Extrapolating to other chromosomes, we expect that a vast majority of blocked zygotes and oocytes probably carry chromosome abnormalities. These data demonstrate the contributions of chromosome disorder in early embryo development blocking and implantation failure. Certainly, the issue of cytoplasm and nuclear immaturity and their relation to each other and to chromosome abnormalities provides a fertile area for future investigation in ART. PMID- 12749039 TI - Chaotic and periodic analysis of fetal magnetocardiogram recordings in growth restriction. AB - OBJECTIVE: We studied how chaotic and periodic heart rate dynamics differ between normal fetuses (n = 19) and intrauterine growth restricted fetuses (n = 11) at 34 to 37 weeks of gestation. We quantified the chaotic dynamics of each heart rate time series obtained by fetal magnetocardiography (FMCG) using correlation dimension. METHODS: The FMCG was recorded digitally by a single-channel biomagnetometer in an electrically shielded room of low magnetic noise. The position of the fetal heart was determined using ultrasonography. RESULTS: The correlation dimension was significantly lower in IUGR than in normal fetuses (p < 0.001, t-test). The periodic dynamics were also obtained by FMCG and measured by power spectrum. The low-frequency components and therefore the periodicity of the low-frequency range were significantly higher in IUGR than in normal fetuses (p < 0.001, t-test). CONCLUSIONS: The analysis of FMCG recordings may offer important perspectives to understand significant features of the heart function of the fetuses. This technique improves the recognition of IUGR fetuses over healthy ones and may help improve perinatal morbidity and mortality. PMID- 12749040 TI - Identification of expressed sequence tags preferentially expressed in human placentas by in silico subtraction. AB - OBJECTIVES: To identify expressed sequence tag (EST) clusters preferentially expressed in placentas. METHODS: The National Center for Biotechnology's online UniGene database contains 14 placenta libraries. In silico (computer-based) subtraction compared placenta libraries against the remaining libraries to identify transcripts preferentially expressed in placentas. For known genes, placental expression or their use in prenatal diagnosis was then explored online using LocusLink and PubMed. RESULTS: Placentas preferentially expressed 475 EST clusters. Of these, 18 EST clusters with no known function were expressed exclusively in placentas. Of the remaining 457 EST clusters, 90 showed preferential placental expression by >/=25 times. Of these 90, literature searches on the 45 EST clusters with known functions showed 44 linked to placental physiology or proposed as markers for prenatal diagnosis [i.e. beta hCG, pregnancy-specific glycoproteins, human placental lactogens, pregnancy associated plasma protein A (PAPP-A)]. Selected genes with known function in pregnancy but whose preferential placental expression fell below the factor of 25 threshold were also identified. CONCLUSION: In silico subtraction identified 44 previously studied genes involved in placental physiology as well as 63 EST clusters preferentially expressed in placental tissue, which may serve as targets for future studies seeking novel markers for prenatal diagnosis or to better understand placental genetics. PMID- 12749041 TI - Using Down syndrome serum screening results to predict low birthweight. AB - OBJECTIVE: To investigate the use of prenatal maternal serum screening results for Down syndrome, for the prediction of low (<2500 g) and very low (<1500 g) birthweight. DESIGN: Record linkage of maternal serum screening results with the corresponding birth records. PARTICIPANTS: 42 259 women whose pregnancies had been screened for the risk of Down syndrome. SETTING: Three East London maternity units, between February 1989 and August 1998. RESULTS: Estimates were made of the effectiveness of single markers only for the prediction of low birthweight, and of multiple markers together with mother's weight and smoking habit. As reported previously, high levels of the single markers alpha-fetoprotein and total human chorionic gonadotrophin, inhibin A, and low levels of unconjugated oestriol were associated with low birthweight. However, the best prediction was obtained when multiple serum markers comprising alpha-fetoprotein, unconjugated oestriol, and inhibin A were used in combination together with mother's weight and adjustment for smoking habit. For a false-positive rate of 5%, this combination predicted 23% of low birthweight and 39% of very low birthweight babies, possibly the best method of prediction to date. CONCLUSION: Prediction of low birthweight derived from Down syndrome screening could be used, for little extra cost, to advise on place of delivery or to select candidates for randomised clinical trials of low birthweight prevention. PMID- 12749042 TI - Enlarged chromosome 13 p-arm hiding a cryptic partial trisomy 6p22.2-pter. PMID- 12749043 TI - Early amnioinfusion for anhydramnios after CVS. PMID- 12749044 TI - Effect of indomethacin on amniotic fluid prostaglandin and aldosterone levels in a fetus with Bartter syndrome. PMID- 12749045 TI - Case report: clinical utility of ultrasound nasal bone determination in the prenatal diagnosis of Down syndrome. PMID- 12749046 TI - Neoplasms associated with alterations in fibroblast growth factor receptors. PMID- 12749047 TI - Rubinstein-Taybi syndrome medical guidelines. AB - Children and adults with Rubinstein-Taybi Syndrome have specific medical conditions that occur with greater frequency than the general population. Based on the available information from the literature and clinical experience, recommendations for specific surveillance and interventions are made to guide those clinicians caring for individuals with Rubinstein-Taybi Syndrome. This is a first attempt at medical guidelines for individuals with RTS in the United States. On-going research is needed in many areas to guide decisions in medical care and allow for refinement of these medical guidelines. PMID- 12749048 TI - Genetic and clinical characterization of patients with an interstitial duplication 15q11-q13, emphasizing behavioral phenotype and response to treatment. AB - The clinical significance of an interstitial duplication of (15)(q11-q13) remains unclear and controversial. The reported phenotypes vary widely and appear to be influenced by the parent of origin of the duplication. Aside from cases of dup(15) reported with autism, the behavioral phenotype of individuals with dup(15) has not been described. We present three families, two with intrachromosomal duplication (15)(q11-q13) ascertained because of developmental delay in a relative. Two families show clear evidence of multigenerational maternal inheritance. The individuals discussed in this paper have minor anomalies and developmental delays. In addition, we describe a behavioral phenotype which often includes attention deficit hyperactivity disorder (ADHD) and autistic spectrum disorder. Responses to medications used to manage these behaviors are also described, including a positive response to methylphenidate and a poor response to fluoxetine. The duplication in each presenting individual, and available family members, was investigated utilizing cytogenetic and molecular techniques including high resolution cytogenetics, fluorescence in situ hybridization (FISH), DNA methylation studies, and quantitative fluorescence PCR. High resolution cytogenetic techniques alone missed some cases, demonstrating the need to confirm results with other methods. PMID- 12749049 TI - Several genomic regions potentially containing QTLs for bone size variation were identified in a whole-genome linkage scan. AB - Bone size is an important determinant of osteoporotic fractures. For a sample of 53 pedigrees that contains more than 10,000 relative pairs informative for linkage analyses, we performed a whole-genome linkage scan using 380 microsatellite markers to identify genomic regions that may contain QTLs of bone size (two dimensional measurement by dual energy X-ray absorptiometry). We conducted two- and multi-point linkage analyses. Several potentially important genomic regions were identified. For example, the genomic region 17q23 may contain a QTL for wrist (ultra distal) bone size variation; a LOD score of 3.98 is achieved at D17S787 in two-point analyses and a maximum LOD score (MLS) of 3.01 is achieved in multi-point analyses in 17q23. 19p13 may contain a QTL for hip bone size variation; a LOD score of 1.99 is achieved at D19S226 in two-point analyses and a MLS of 2.83 is achieved in 19p13 in multi-point analyses. The genomic region identified on chromosome 17 for wrist bone size seems to be consistent with that identified for femur head width variation in an earlier whole-genome scan study. The genomic regions identified in this study and an earlier investigation on one-dimensional bone size measurement by radiography are compared. The two studies may form a basis for further exploration with larger samples and/or denser markers for confirmation and fine mapping studies to eventually identify major functional genes and the associated etiology for osteoporosis. PMID- 12749050 TI - IL-1 alpha (-889) promoter polymorphism is a risk factor for osteomyelitis. AB - As osteomyelitis (OM) induces the synthesis of inflammatory cytokines and IL-1 mediates bone resorption by osteoclasts we determined if there is an association between certain common polymorphisms of the genes encoding proinflammatory cytokines (IL-1 alpha and beta, IL-6, TNF-alpha) and OM in adults. The IL-1 alpha (-889) TT genotype was significantly more frequent among 52 OM patients than in 109 healthy controls (13/52, [25.0%] vs. 9/109, [8.3%], P = 0.0081, chi(2) = 7.01, OR = 3.7, 95% CI, 1.35-10.34). Patients who were homozygous for the T allele were younger than the rest of the OM patients (mean age 35.7 +/- 11.5 vs. 58.1 +/- 18.6 years, P = 0.001). IL-1 beta TT (+3953) polymorphism was also more frequent in OM patients (P = 0.014, chi(2) = 5.12, OR = 5.1, 95% CI, 1.21-52.14), but IL-1 beta is in linkage disequilibrium with the IL-1 alpha *T (P < 0.001). Route of infection, chronicity of the infection, type of microorganism isolated, and frequency of relapses were similar in patients with and without the IL-1 alpha TT genotype. There were no associations between OM and polymorphisms of other cytokines genes. IL-1 alpha serum levels were significantly increased in all the OM patients independently of their IL-1 genotype compared to the controls (P = 0.021). Although IL-1 alpha serum levels were not significantly higher in patients with the IL-1 alpha (-889) polymorphism, this does not exclude a difference in production of IL-1 alpha by osteoclasts or other inflammatory cells at the site of infection. PMID- 12749051 TI - Food fortification with folic acid and twinning among California infants. AB - This study sought to determine whether a change in twinning prevalence was associated with mandatory folic acid fortification. As of January 1998, it became mandatory in the United States that grain products be fortified with folic acid. The effectiveness of this fortification for neural tube defects has been explored, but other possible implications of fortification has not. We computed the prevalence of twinning among more than 2.5 million California births occurring from 1990 to 1999. The prevalence of twin births in California increased over the last decade with increases observed among most maternal race/ethnic, age, and parity groupings. Unadjusted prevalence measures did not reveal substantial increases in twinning prevalence among these groupings after the fortification period relative to the period immediately preceding it. Analyses that simultaneously adjusted for age, parity, race/ethnicity, sexes of twin pair (as a measure of zygosity), year of birth, and fortification period also did not reveal an association between fortification and twinning. Our results do not suggest a change in twinning prevalence associated with folic acid fortification of the US food supply among women delivering in California. PMID- 12749053 TI - Low penetrance of the 14484 LHON mutation when it arises in a non-haplogroup J mtDNA background. AB - The penetrance in Leber's hereditary optic neuropathy (LHON) pedigrees is determined primarily by a mutation in the mitochondrial genome (mtDNA), but secondary factors are also necessary for manifestation of the disorder. It has been proposed that mtDNA polymorphisms affect penetrance in LHON pedigrees. In particular, it has been postulated that one or more polymorphisms associated with European haplogroup J mtDNAs substantially increase the penetrance of the primary LHON mutation at nucleotide 14484. We report here a haplogroup H matrilineal pedigree (VIC14) in which the single affected member carries the 14484 LHON mutation, but who manifested a milder and atypical optic nerve disorder. In addition, during a population screen, we identified an individual who carried the 14484 mutation but who had normal vision. Finally, the 14484 mutation is under represented among haplogroup H mtDNAs that carry a LHON mutation. These results, in conjunction with other studies that are reviewed, indicate that 14484 LHON mutations have a low penetrance when they arise in a haplogroup H mtDNA background. PMID- 12749054 TI - Ethanol consumption and DRD2 gene TaqI a polymorphism among socially drinking males. AB - The dopaminergic system in the human brain is thought to play a major role in the development of alcohol consumption habits and alcoholism. It has been reported that homozygous D2-/- knock-out mice lacking D2 receptors consume about 50% to 60% less ethanol than wild-type D2+/+ mice, and heterozygous mice have an intermediate level of alcohol consumption. The DRD2 gene TaqI A polymorphism has been suggested to associate with a low D2 receptor density in post mortem and in vivo measurements. Numerous association studies on this polymorphism and alcoholism have shown most controversial results. We studied whether DRD2 TaqI A genotype affects alcohol consumption in an ethnically homogeneous, representative sample of 1,019 Finnish Caucasian males. After excluding the abstainers from the study, the self-reported alcohol consumption among the remaining 884 non abstainers was compared in the TaqI A genotype groups (A1/A1, A1/A2, A2/A2). The alcohol consumption of the homozygous A1/A1 group was about 30% lower than in A1/A2 group, and 40% lower than in A2/A2 group (P = 0.042 and 0.041 in a sociodemographic variable-adjusted multivariate model). The results indicate an association between DRD2 genotype and alcohol consumption habits in humans. These results in the large sample of non-alcoholic males are also opposite to some previous findings on the higher A1 allele frequency among alcoholic populations. PMID- 12749055 TI - What does "a gene for heart disease" mean? A focus group study of public understandings of genetic risk factors. AB - There is growing concern in the medical community about potential genetic determinism in the patient population. Limited information about the public understanding of genetic factors in disease formation is available. To access public perceptions of potentially deterministic phrasing of genetic risk factors, we sought to establish interpretations of the phrase, "a gene for heart disease." Focus groups in urban, suburban, and rural communities were conducted from July through October, 2001 in Georgia. A total of 108 participants were recruited. Participants were recruited to balance sex and racial representation. We used three outcome measures for participants understandings of the phrase: (1) participants' statements of the meaning of the phrase; (2) the level of determinism assigned to genetic factors by participants; and (3) participant reports of the health consequences of having "a gene for heart disease." Participants did not report a single interpretation of the phrase. There were dominant participant interpretations under each outcome measure: (1) "a gene for heart disease" was interpreted as meaning genetic and environmental factors both played roles in disease formation; (2) genetic predisposition was perceived as heightened, not absolute, risk; (3) the perceived health impact was a greater risk of becoming sick. Minority interpretations were found under each measure. Overall, naming "a gene for heart disease" does not appear to have a deterministic impact on a plurality of participants' perceptions of risks associated with genetic factors. Genetic fatalism in patient populations may be confined to a sizable minority. Important considerations for provider intervention and patient education are indicated. PMID- 12749052 TI - Mortality and apolipoprotein E in African-American, and White elders: an attempted replication. AB - We have tried, with only partial success, to confirm findings in a recently reported study in this journal on the relationship of APOE genotype to mortality in community representative Hispanics (n = 659), Whites (n = 272), and African Americans (n = 450), aged 65 and over, living in Northern Manhattan, New York. That study found that using proportional hazards models adjusted for sex and lipid levels, Hispanics and Whites with the E2/E3 genotype, but not African Americans, had the lowest mortality risk. Those under age 75 had risks comparable to those over age 75, suggesting minimal survivor bias. Nearly 50% of the mortality risk associated with the APOE genotype appeared to act through heart disease, diabetes, and stroke. The current study of African-Americans (n = 1,083) and Whites (n = 915) aged 71 and over living in the more rural Southeastern US, found no protective effect of the E2/E3 genotype for either African-Americans or Whites. Among younger Whites (age 71-75), point estimates suggested that the E2/E3 genotype might be protective, but at a nonsignificant level; self-reported African-American race, but not genotype, was a risk factor for mortality in this age group. Neither lipid level nor health condition attenuated the effect of APOE genotype. Differences in findings may reflect issues of sampling, age, the relative distribution of the APOE alleles, or some other factor. Until such time as studies use truly representative samples and include younger ages, findings in this area must be treated with caution. PMID- 12749056 TI - Isolated noncompaction of the left ventricular myocardium in the adult is an autosomal dominant disorder in the majority of patients. AB - Isolated noncompaction of the ventricular myocardium (INVM, MIM 300183 and 604169) is a congenital unclassified cardiomyopathy with numerous prominent trabeculations and deep intertrabecular recesses in a hypertrophied and hypokinetic myocardium. Mutations in the G4.5 gene result in a wide spectrum of severe infantile X-linked cardiomyopathic phenotypes including Barth syndrome with dilated cardiomyopathy and INVM. Molecular genetic analysis of INVM has only been performed in pediatric patients. Although adult INVM patients show similar cardiac abnormalities, the influence of genetic factors, especially of mutations in G4.5, is unknown. We analyzed 25 adult INVM patients for the presence of mutations in the G4.5 gene and performed a pedigree analysis of probands. Mutations were not found in the coding sequence or splice sites of G4.5. Systematic analysis of relatives from seven of nine probands showed multiple affected members consistent with an autosomal dominant pattern of inheritance in the majority of cases. We conclude that INVM in the adult is an autosomal dominant disorder rarely caused by mutations in G4.5 and therefore genetically distinct from infantile X-linked cases. PMID- 12749057 TI - Coenzyme Q10 levels in Prader-Willi syndrome: comparison with obese and non-obese subjects. AB - Coenzyme Q10 (CoQ10) is an essential component of the mitochondrial respiratory chain and an important scavenger of reactive oxygen species. Low levels are found in individuals with reduced energy expenditure, cardiac and skeletal muscle dysfunction, and mitochondrial disorders, many of these manifestations are seen in individuals with Prader-Willi syndrome (PWS). In addition, CoQ10 supplementation frequently is given to individuals with this syndrome. To determine if CoQ10 levels are decreased in PWS, we studied plasma CoQ10 levels in 16 subjects with PWS, 13 with obesity of unknown cause, and 15 subjects without obesity but of similar age and compared with body composition. Plasma CoQ10 levels were significantly decreased (P < 0.05), using several statistical approaches in subjects with PWS (0.45 +/- 0.16 microg/ml), compared to subjects without obesity (0.93 +/- 0.56 microg/ml), but not different from subjects with obesity (0.73 +/- 0.53 microg/ml). When plasma CoQ10 was normalized relative to cholesterol, triglyceride, and creatinine levels and fat and lean mass [determined by dual energy X-ray absorptiometry (DEXA)] in the subjects with either PWS or obesity, no significant differences were observed. However, a lower muscle mass was found in the PWS subjects. PMID- 12749058 TI - Common variant in betaine-homocysteine methyltransferase (BHMT) and risk for spina bifida. AB - Neural tube defects (NTD) are common malformations resulting from incomplete closure of the neural tube in the first month after conception. Since genetic deficiencies in folate-dependent homocysteine metabolism have been identified in NTD families, we investigated a common variant in betaine-homocysteine methyltransferase (BHMT), 742G-->A (R239Q), as a genetic modifier of NTD risk. Genotypes, nutrient levels, and plasma total homocysteine (tHcy) were assessed in 54 patients with spina bifida, 57 mothers of patients, 93 control children, and 86 mothers of controls. The QQ genotype (present in 17% and 7% of the control and case mothers, respectively, and in 12% and 6% of the control and case children, respectively) was associated with a decreased risk of NTD (odds ratios of 0.52 (95% CI 0.13-2.05) for children and 0.37 (95% CI 0.11-1.22) for mothers). The small sample size limited the statistical power of the analyses, but these decreases, although not statistically significant, are compatible with a protective effect. We did not observe statistically-significant genotype dependent differences in plasma homocysteine, although women with the QQ genotype did have lower homocysteine; in children, the mean homocysteine level was higher in the QQ group. This inconsistency could be explained by the fact that age is a strong determinant of homocysteine in children and the QQ group was on average older than the other genotype groups. Our study suggests that the Q allele of the R239Q mutation may decrease risk of the condition. This warrants further investigation of its relationship with the development of NTD. PMID- 12749059 TI - Absence of 12q21.2q22 deletions and subtelomeric rearrangements in cardiofaciocutaneous (CFC) syndrome patients. AB - Recent publications described two patients with a CFC-like phenotype and the same deletion of chromosome region 12q21.2q22 [Rauen et al., 2000, 2002]. The patients did not have the classical CFC phenotype and presented other signs not usually seen in CFC patients: the first patient had hydrocephalus, and the second, a history of olygohydramnios, normal stature, pyloric stenosis, cutaneous syndactyly of toes and bilateral transverse palmar creases. In order to verify if classic CFC patients with normal chromosomes in conventional preparations have microdeletions within the 12q21.2q22 chromosome region, we performed FISH analysis using 12 BAC probes to screen this area. The average interval between the probes was of approximately 1 Mb. No deletions were found in any of the 17 classical CFC patients we examined. We conclude that the region 12q21.2q22 is not a candidate region for CFC syndrome and that the patients described by Rauen et al. [2000, 2002] probably have a different condition, i.e., an aneuploidy syndrome, with some phenotypic resemblance to the CFC syndrome. To further evaluate the possibility of other chromosome imbalances, we performed a subtelomeric analysis, by FISH technique, of all chromosomes, and did not find any subtelomeric rearrangements. PMID- 12749060 TI - Angelman syndrome associated with oculocutaneous albinism due to an intragenic deletion of the P gene. AB - Angelman syndrome (AS) is a neurodevelopmental disorder characterized by mental retardation, speech impairment, ataxia, and happy disposition with frequent smiling. AS results from the loss of expression of a maternal imprinted gene, UBE3A, mapped within 15q11-q13 region, due to different mechanisms: maternal deletion, paternal UPD, imprinting center mutation, and UBE3A mutation. Deletion AS patients may exhibit hypopigmentation of skin, eye, and hair correlating with deletion of P gene localized in the distal part of Prader-Willi (PWS)/AS region. Our patient presented developmental delay, severe mental retardation, absence of speech, outbursts of laughter, microcephaly, ataxia, hyperactivity, seizures, white skin, no retinal pigmentation, and gold yellow hair. His parents were of African ancestry. The SNURF-SNRPN methylation analysis confirmed AS diagnosis and microsatellite studies disclosed deletion with breakpoints in BP2 and BP3. All of the 25 exons and flanking introns of the P gene of the patient, his father, and mother were investigated. The patient is hemizygous for the deleted exon 7 of the P gene derived from his father who is a carrier of the deleted allele. Our patient manifests OCA2 associated with AS due to the loss of the maternal chromosome 15 with the normal P allele, and the paternal deletion in the P gene. As various degrees of hypopigmentation are associated with PWS and AS patients, the study of the P gene in a hemizygous state could contribute to the understanding of its effect on human pigmentation during development and to disclose the presence of modifier pigmentation gene(s) in the PWS/AS region. PMID- 12749061 TI - Association of microphthalmia and esophageal atresia: description of a patient and review of the literature. AB - Since 1988, when Rogers first described a boy with anophthalmia associated with esophageal atresia, eight similar cases have been reported. These patients lend support to the hypothesis that this association of congenital anomalies constitutes a discrete entity, although the etiology is still unknown. We report a patient with this combination of malformations as well as a marked hypoplasia of the entire left half of the body. PMID- 12749062 TI - Melorheostosis in a family with autosomal dominant osteopoikilosis: report of a third family. AB - We describe a three-generation family with clinical and radiological findings of osteopoikilosis in five and melorheostosis in one individual. The co-occurrence of both rare bone disorders suggests that both conditions might be related as suggested previously by Butkus et al. [1997: Am J Med Genet 72:43-46] and Nevin et al. [1999: Am J Med Genet 82:409-414]. The findings in this family strengthen the hypothesis that osteopoikilosis is an autosomal dominant condition and that an early postzygotic second hit mutation in the second allele results in melorheostosis. PMID- 12749063 TI - Malignant hematological disorders in children with Wolf-Hirschhorn syndrome. AB - Wolf-Hirschhorn syndrome (WHS) is a rare chromosomal disorder attributable to a deletion at the short arm of chromosome 4. This syndrome is associated with characteristic facial appearance, multiple congenital abnormalities, mental retardation, feeding difficulties and failure to thrive. We report two girls with WHS who developed myelodysplastic syndrome (MDS). According to the "Category, Cytology, Cytogenetic (CCC)"classification of childhood MDS, patient 1 had refractory cytopenia with ring sideroblasts at the age of 6 years, while patient 2 had refractory cytopenia with dysplasia at the age of 5-1/2 years. Patient 1 progressed to refractory cytopenia with excess blasts within a year, while patient 2 progressed to acute lymphoblastic leukemia within 1 month of presentation. It is possible that allelic loss of a tumor suppressor gene such as WHSC1 and/or FGFR3 from the deleted segment 4p16.3 plays a critical role in the process of malignant transformation. To our knowledge, this is the first report of severe hematological complications like MDS and leukemia in children with WHS and may be an important genetic model for understanding malignant hematological transformation. This report also underscores the importance of evaluating children with WHS for hematopoietic dysfunction. PMID- 12749064 TI - New syndrome of simplified gyral pattern, micromelia, dysmorphic features and early death. AB - We report two sisters with a new syndrome of simplified gyral pattern, normal head circumference at birth but with subsequent development of microcephaly, intractable seizures, and early death. Dysmorphic features included coarse face, hypertrichosis, short nose, paranasal widening, long philtrum, short neck, upper limb micromelia, single transverse palmar lines, and clasp thumbs. The proband had repeated convulsions from shortly after birth and she required continuous artificial ventilation. Neurological examination showed absent sucking, rooting, Moro and grasping reflexes. MRI revealed a diffuse simplified gyral pattern with apparent agyria over the frontal lobes. Biochemical screening gave normal results. Her older sister had bilateral renal pelvic dilatation on prenatal ultrasound. She also developed severe convulsions on the first day of life, and she had to be artificially ventilated for 38 days. She had severe developmental retardation and neurological examination showed absence of spontaneous movements and Moro reflex, weak sucking reflex, and hypertonicity. CT scan of the brain showed a simplified gyral pattern. At 3 months, she developed hypocalcemia and hyperphosphatemia with normal levels of vitamin D and alkaline phosphatase, and parathyroid hormone level was low. Other biochemical tests gave normal results. She died at 5 months due to a massive aspiration event. Based on the unique clinical and radiological features found in our patients, we propose that this is a new syndrome. PMID- 12749065 TI - Subcortical band heterotopia with simplified gyral pattern and syndactyly. AB - We describe a girl with an unusual form of subcortical band heterotopia (SBH) and a complex malformation syndrome. SBH had an irregular inner margin, organized in contiguous fascicles of migrating neurons, sometimes giving the appearance of many small contiguous gyri. The true cortex had decreased thickness and showed a simplified gyral pattern with decreased number of gyri, which were usually of increased width, and shallow sulci. The cerebellum was hypoplastic. Additional features included epicanthal folds, hypertelorism, small nose with hypoplastic nares, bilateral syndactyly of the toes, pulmonary valve stenosis, atrial and ventricular septal defects. At the age of 1 year the patient had severe developmental delay and epilepsy. Chromosome studies and mutation analysis of the DCX and LIS1 genes gave negative results. This observation delineates a new multiple congenital abnormalities mental retardation syndrome and confirms genetic heterogeneity of SBH. PMID- 12749066 TI - Unusual congenital abdominal wall defect and review. AB - Abdominal wall defects distant from the umbilicus are very unusual. We describe a patient with a congenital defect adjacent and external to the lateral border of the recti muscles, rounded in shape, bilateral and symmetrical, but affecting different structures in each side, musculocutaneous agenesis in the left side and only muscle agenesis in the right one. The morphologic characteristics of the defect and the absence of associated anomalies, suggest that this anomaly may be the result of an event acting during phenogenesis. We consider that this defect is different from other structural abnormalities of the anterior abdominal wall which occur much earlier at certain periods of the embryogenesis. Though the etiology is speculative, we could postulate a disruptive origin of the defect of unknown cause. PMID- 12749067 TI - Primary hypergonadotropic hypogonadism, partial alopecia, and Mullerian hypoplasia: report of a second family with additional findings. AB - Two sisters with primary hypergonadotropic hypogonadism associated with microcephaly, flat occiput, partial alopecia, absent or streak ovaries, and Mullerian hypoplasia are reported. Their parents are first cousins. Despite some clinical differences, their features were very close to a family described with such an association by Al-Awadi et al. [1985: Am J Med Genet 22:619-622] in Kuwait. PMID- 12749068 TI - Osseous fragility in Marshall-Smith syndrome. AB - Marshall-Smith syndrome is characterized by accelerated osseous maturation, craniofacial anomalies, failure to thrive, psychomotor delay, hypotonia, pulmonary dysfunction, and limited life expectancy. We describe a 7-year-old girl who, in addition to meeting these criteria for Marshall-Smith syndrome, had multiple fractures and skeletal anomalies. The purpose of this report is to draw attention to Marshall-Smith syndrome as one of the skeletal dysplasias characterized by osseous fragility. PMID- 12749069 TI - Polysyndactyly, complex heart malformations cardiopathy, and hepatic ductal plate anomalies: an autosomal recessive syndrome diagnosed antenatally. AB - A distinct syndrome was ascertained in a 3-year-old girl and her brother. The proband was the first child of first cousin parents. She was born after an uneventful pregnancy. At birth, multiple congenital anomalies were noted: ptosis of the left eyelid, hypertelorism, anteverted nares, large fontanel, long philtrum, ungueal hypoplasia, polysyndactyly, single transverse crease, complex cardiopathy, and hepatic cysts. During another pregnancy of the mother, fetal ultrasonographic examination showed an hypertrophy of the right ventricle and atria, a dextroposition of the aorta, a bilateral renal pelvis dilatation, and a club foot. After termination of the pregnancy, necropsy showed facial anomalies, a small penis, a polysyndactyly, a ventricular septum defect, and a malformation of the ductal plate. Bonneau et al. [1983: J Genet Hum 2:93-105] described a family in which three sibs had a complex cardiac malformation, hexadactyly of the first toe, and syndactyly of the third and fourth fingers. Rajab [1997: Clin Dysmorphol 6:85-88] described two sibs with similar features in an Omani family. The sibs described in this report had anomalies of the ductal plate which were not reported in the two other families. These new findings are in favor of autosomal inheritance of this condition which is amenable to antenatal diagnosis. PMID- 12749070 TI - Nevi flammei affecting two contralateral quadrants and nevus depigmentosus: a new type of phacomatosis pigmentovascularis? AB - Here, we report on a 64-year-old-woman with nevi flammei affecting two contralateral quadrants, venous insufficiency of the right leg, and asymmetry of the arms, in association with a linear nevus depigmentosus. This combination of vascular and pigmentary abnormalities may represent a novel type of phacomatosis pigmentovascularis caused by non-allelic twin-spotting. PMID- 12749071 TI - Chiari I malformation and cloacal exstrophy: report of a patient with both defects of blastogenesis. AB - We report on child with Chiari I malformation (CIM) and cloacal exstrophy, a combination of findings that has not been reported previously. CIM and cloacal exstrophy both demonstrate abnormalities that represent maldevelopment of the midline field. This combination of anomalies in this patient suggests an impairment of midline development during blastogenesis. PMID- 12749072 TI - Male patient with non-mosaic deleted Y-chromosome and clinical features of Turner syndrome. AB - Turner syndrome is hypothesized to result from haplo-insufficiency of a gene or perhaps multiple genes present on the sex chromosomes; however, the frequent association of mosaicism with deletions of the sex chromosomes prevents establishing useful genotype/phenotype correlations. In this clinical report, we present a male with a de novo, non-mosaic deletion of the Y-chromosome. The phenotype of this patient is unlike any similar cases previously reported in the literature. This patient exhibits many classical clinical features of Turner syndrome including short stature, characteristic facial anomalies, and webbed neck with low posterior hairline, aortic valve abnormality, and hearing impairment. Detailed molecular characterization of this deleted Y-chromosome could provide important information towards establishing genotype/phenotype correlations in Turner syndrome. PMID- 12749073 TI - Acrofrontofacionasal dysostosis: report of the third Brazilian family. PMID- 12749074 TI - Association study of autistic disorder and chromosome 16p. PMID- 12749075 TI - Successful carnitine therapy for Raynaud's phenomenon in velo-cardio-facial syndrome. PMID- 12749078 TI - Molecular composition and surface properties of storage lipid particles in wax bean (Phaseolus vulgaris). AB - Lipid particles have been isolated from seeds of wax bean (Phaseolus vulgaris), a species in which starch and protein rather than lipid are the major seed storage reserves. These lipid particles resemble oil bodies present in oil-rich seeds in that > 90% of their lipid is triacylglycerol. Moreover, this triacylglycerol is rapidly metabolized during seed germination indicating that it is a storage reserve. The phospholipid surfaces of oil bodies are known to be completely coated with oleosin which prevents their coalescence, particularly during desiccation of the developing seed. This would appear to be necessary since lipid is the major storage reserve in oil seeds, and there are very few alternate types of storage particles in the cytoplasm of oil seed endosperm to provide a buffer against coalescence of oil bodies by isolating them from one another. The present study indicates that the surfaces of lipid particles from wax bean are not completely coated with oleosin and feature regions of naked phospholipid. This finding has been interpreted as reflecting the fact that lipid particles in wax been seeds are less prone to coalescence than oil bodies of oil-rich seeds. This arises because the individual lipid particles are interspersed in situ among highly abundant protein bodies and starch grains and hence less likely to come in contact with one another, even during desiccation of the developing seed. PMID- 12749080 TI - Crystallochemical characterization of calcium oxalate crystals isolated from seed coats of Phaseolus vulgaris and leaves of Vitis vinifera. AB - Calcium oxalate crystals are a major biomineralization product in higher plants. Their biological function and use are not well understood. In this work, we focus on the isolation and crystallochemical characterization of calcium oxalate crystals from seed coats of Phaseolus vulgaris (prisms) and leaves of Vitis vinifera (raphides and druses) using ultrastructural methods. A proposal based on crystal growth theory was used for explaining the existence of different morphologies shown by these crystals grown inside specialized cells in plants. PMID- 12749079 TI - Coumarin inhibits the growth of carrot (Daucus carota L. cv. Saint Valery) cells in suspension culture. AB - We used a carrot (Daucus carota L. cv. Saint Valery) cell suspension culture as a simplified model system to study the effects of the allelochemical compound coumarin (1,2 benzopyrone) on cell growth and utilisation of exogenous nitrate, ammonium and carbohydrates. Exposure to micromolar levels of coumarin caused severe inhibition of cell growth starting from the second day of culture onwards. At the same time, the presence of 50 mumol/L coumarin caused accumulation of free amino acids and of ammonium in the cultured cells, and stimulated their glutamine synthetase, glutamate dehydrogenase, glucose-6-phosphate dehydrogenase and phosphoenolpyruvate carboxylase activities. Malate dehydrogenase, on the other hand, was inhibited under the same conditions. These effects were interpreted in terms of the stimulation of protein catabolism and/or interference with protein biosynthesis induced by coumarin. This could have led to a series of compensatory changes in the activities of enzymes linking nitrogen and carbon metabolism. Because coumarin seemed to abolish the exponential phase and to accelerate the onset of the stationary phase of cell growth, we hypothesise that such allelochemical compounds may act in nature as an inhibitor of the cell cycle and/or as a senescence-promoting substance. PMID- 12749081 TI - Age-dependent response of maize leaf segments to cadmium treatment: effect on chlorophyll fluorescence and phytochelatin accumulation. AB - The relationship between the age of leaf tissue and response of the photosynthetic apparatus and phytochelatin accumulation to Cd treatment was studied. Studies were carried out with seedlings of Zea mays L. cv. Hidosil grown in the presence of 100-200 mumol/L Cd for 14 days under low light conditions. The third leaf was divided into 3 segments of equal length differing in the stage of tissue maturity and used for measurements of chlorophyll content, chlorophyll fluorescence, glutathione and phytochelatin content and Cd accumulation. A close relationship between the age of leaf tissue and response of the photosynthetic apparatus to Cd was shown. Cadmium (200 mumol/L) reduced photochemical processes more in older than younger leaf segments as seen in the Chl fluorescence parameters Fv/F0, and t1/2, while the chlorophyll fluorescence decrease ratio (Rfd) was inhibited more strongly in younger ones. Fv/Fm was slightly affected. Cd-induced enhancement of GSH content was correlated with higher phytochelatin accumulation to a greater extent in younger than in older leaf segments. Phytochelatin level corresponded to changes of photochemical processes in older leaves. The peptide thiol:Cd molar ratio for the phytochelatins varied depending on Cd concentration and age of leaf segments. The protective role of phytochelatins for the photosynthetic apparatus is discussed. PMID- 12749082 TI - Induction of myrosinase gene expression and myrosinase activity in radish hypocotyls by phototropic stimulation. AB - The role of myrosinase (beta-thioglucoside glucohydrolase, EC 3.2.3.1) in the phototropic response in radish hypocotyls was investigated. Unilateral illumination with blue light abruptly up-regulated the activity of myrosinase, which releases bioactive 4-methylthio-3-butenyl isothiocyanate (MTBI) from inactive 4-methylthio-3-butenyl glucosinolate (MTBG), in the illuminated halves of radish hypocotyls 10 min after onset of phototropic stimulation, peaking after 30 min and decreasing thereafter. The myrosinase activity in the shaded halves also increased, but was significantly lower than that in the illuminated halves. Furthermore, whether blue light illumination induces myrosinase gene expression was studied. Northern blotting analysis indicated that myrosinase mRNA levels were increased markedly in unilaterally illuminated hypocotyls, reaching maximum signal intensity within 10 min after onset of blue illumination, declining nearly to the control level thereafter. These results suggested that phototropic stimulation promotes myrosinase gene expression and myrosinase activity in the illuminated side, resulting in the conversion of inactive MTBG to active MTBI and simultaneously producing more active raphanusanins, causing a phototropic response. PMID- 12749083 TI - Pollination-induced ethylene promotes the early phase of pollen tube growth in Petunia inflata. AB - In Petunia inflata, a species with gametophytic self-incompatibility, pollination triggers two phases of ethylene production by the pistil, the first of which peaks 3 hours after pollination with compatible or incompatible pollen. To investigate the physiological significance of the first phase of ethylene production, pollinated flowers were treated with 2,5-norbornadiene (NBD), an inhibitor of ethylene action. Treatment with NBD reduced pollen tube growth in a dose-dependent manner during the first six hours after pollination; however, pollen tube growth was insensitive to NBD if the treatment was applied 6 hours or more after pollination. Simultaneous application of exogenous ethylene substantially offset the inhibitory effects of NBD in flowers pollinated for 4 hours. Another inhibitor of ethylene action, 1-methylcyclopropene (1-MCP), also produced a strong inhibition of pollen tube growth during the first six hours of pollination. The experiments with 1-MCP pretreatment indicate that pistil tissues are the primary target of the pollination-induced ethylene. PMID- 12749084 TI - Relationships between leaf chlorophyll content and spectral reflectance and algorithms for non-destructive chlorophyll assessment in higher plant leaves. AB - Leaf chlorophyll content provides valuable information about physiological status of plants. Reflectance measurement makes it possible to quickly and non destructively assess, in situ, the chlorophyll content in leaves. Our objective was to investigate the spectral behavior of the relationship between reflectance and chlorophyll content and to develop a technique for non-destructive chlorophyll estimation in leaves with a wide range of pigment content and composition using reflectance in a few broad spectral bands. Spectral reflectance of maple, chestnut, wild vine and beech leaves in a wide range of pigment content and composition was investigated. It was shown that reciprocal reflectance (R lambda)-1 in the spectral range lambda from 520 to 550 nm and 695 to 705 nm related closely to the total chlorophyll content in leaves of all species. Subtraction of near infra-red reciprocal reflectance, (RNIR)-1, from (R lambda)-1 made index [(R lambda)(-1)-(RNIR)-1] linearly proportional to the total chlorophyll content in spectral ranges lambda from 525 to 555 nm and from 695 to 725 nm with coefficient of determination r2 > 0.94. To adjust for differences in leaf structure, the product of the latter index and NIR reflectance [(R lambda)( 1)-(RNIR)-1]*(RNIR) was used; this further increased the accuracy of the chlorophyll estimation in the range lambda from 520 to 585 nm and from 695 to 740 nm. Two independent data sets were used to validate the developed algorithms. The root mean square error of the chlorophyll prediction did not exceed 50 mumol/m2 in leaves with total chlorophyll ranged from 1 to 830 mumol/m2. PMID- 12749085 TI - Electrolyte leakage and lipid degradation account for cold sensitivity in leaves of Coffea sp. plants. AB - Five Coffea genotypes differing in their sensitivity to low positive temperatures were compared with regard to the effects of chilling on membrane integrity, as well as their ability to recover from cold-induced injury upon re-warming. Membrane damage was evaluated through electrolyte leakage, changes in membrane lipid composition and malondialdehyde (MDA) production in control conditions (25/20 degrees C, day/night), after a gradual temperature decrease period to 15/10 degrees C, after chilling treatment (3 nights at 4 degrees C) and upon re warming to 25/20 degrees C during 6 days (recovery). C. dewevrei showed the highest electrolyte leakage at 15/10 degrees C and after chilling. This was due mainly to lipid degradation observed at 15/10 degrees C, reflecting strong membrane damage. Furthermore, MDA production after chilling conditions indicated the occurrence of lipid peroxidation. A higher susceptibility of C. dewevrei to cold also was inferred from the complete absence of recovery as regards permeability, contrary to what was observed in the remaining plants. Apoata and Piata presented significant leakage values after chilling. However, such effects were reversible under recovery conditions. Exposure to cold (15/10 degrees C and 3 x 15/4 degrees C) did not significantly affect membrane permeability in Catuai and Icatu. Furthermore, no significant MDA production was observed even after chilling treatments in Apoata, Piata, Catuai and Icatu, suggesting that the four genotypes had the ability to maintain membrane integrity and/or repair membrane damage caused by low temperatures. Apoata, Piata and, to a lower extent, Catuai, were able to cope with gradual temperature decrease through an enhanced lipid biosynthesis. After acclimation, Piata and Catuai showed a lowering of digalactosyldiacylglycerol to monogalactosyldiacylglycerol ratio (MGDG/DGDG) as a result of enhanced DGDG synthesis, which represents an increase in membrane stability. The same was observed in Apoata after chilling, in spite of phospholipids decrease. The studied parameters clearly indicated that chilling induced irreversible membrane damage in C. dewevrei. We also concluded that increased lipid synthesis, lower MGDG/DGDG ratio, and changes in membrane unsaturation occurring during acclimation to low temperatures may be critical factors in maintenance of cellular integrity under chilling. PMID- 12749087 TI - cDNA-AFLP display for the isolation of differentially expressed genes during chicory root development. AB - To identify genes expressed during root enlargment during the early vegetative growth of chicory (Cichorium intybus L.), we used the cDNA-AFLP technology. The radial pattern of chicory roots was investigated by histological analysis to determine the spatial vascular cambium setting. In young plantlets, serial root sections showed that differentiation of secondary tissues occurred along a gradient extending from the apex to the crown. The cDNA-AFLP technique was carried out on total mRNAs extracted from root tissues producing secondary structures and root tissues developing primary structures only. This study reports on the isolation of two transcript-derived fragments (TDFs) referred to as Y-16 and Y-21. Sequence analysis at the protein level showed that Y-16 carries a sequence highly homologous (93% identities) to the amino acid transporter-like protein 1 (AATL 1) from Arabidopsis and Y-21 presents 72% identity to AAD25141.1 Arabidopsis protein. The expression pattern of both these TDFs was analysed by northern blot and showed an over-expression during early development of chicory roots. PMID- 12749086 TI - Up-regulation of glutathione metabolism and changes in redox status involved in adaptation of reed (Phragmites communis) ecotypes to drought-prone and saline habitats. AB - The glutathione (GSH) metabolic characteristics and redox balance in three ecotypes of reed (Phragmites communis), swamp reed (SR), dune reed (DR), and heavy salt meadow reed (HSMR), from different habitats in desert regions of northwest China were investigated. The DR possessed the highest rate of GSH biosynthesis and metabolism with the lowest levels of total and reduced GSH and its biosynthetic precursors, gamma-glutamylcysteine (gamma-EC) and cysteine (Cys), of the three reed ecotypes. This suggests that a higher rate of GSH biosynthesis and metabolism, but not GSH accumulation, might be involved in the adaptation of this terrestrial reed ecotype to its dry habitat. The HSMR shared this profile although it exhibited the highest reduced thiol levels of the three ecotypes. Two key enzymes in the Calvin-cycle possessing exposed sulfhydryl groups, NADP(+)-dependent glyceraldehydes-3-phosphate dehydrogenase (G3PD) and fructose-1,6-bisphosphatase (FBPase), and other two key enzymes in the pentose phosphate pathway (PPP), glucose-6-phosphate dehydrogenase (G6PDH) and 6 phosphogluconate dehydrogenase (6-PGD), had very similar activities in the three reed ecotypes. Compared to the SR, the DR and HSMR had higher ratios of NADPH/NADP+ and NADH/NAD+, indicating that a more reduced redox status in the plant cells might be involved in the survival and adaptation of the two terrestrial reed ecotypes to long-term drought and salinity, respectively. These results suggest that changes of GSH metabolism and redox balance were important components of the adaptation of reed, a hydrophilic plant, to more extreme dune and saline habitats. The coordinated up-regulations of the rate of GSH biosynthesis and metabolism and reduction state of redox status of plant cells, conferred on the plant high resistance or tolerance to long-term drought and salinity. PMID- 12749089 TI - Chloroplasts in seeds and dark-grown seedlings of lotus. AB - In most higher plants, mature dry seeds have no chloroplasts but etioplasts. Here we show that in a hydrophyte, lotus (Nelumbo nucifera), young chloroplasts already exist in shoots of mature dry seeds and that they give rise to mature chloroplasts during germination, even in darkness. These shoots contain chlorophyll and chlorophyll-binding proteins CP1 and LHCP. The unique features of chloroplast formation in N. nucifera suggest a unique adaptive strategy for seedling development correlated with the plant's habitat. PMID- 12749088 TI - Metabolic and ultrastructural responses of lupine embryo axes to sugar starvation. AB - Embryo axes isolated from germinating lupine seeds were cultivated in vitro for 24-96 h over media containing either 60 mmol/L sucrose or no sucrose. Ultrastructural studies showed that large vacuoles were accumulating in a central region of primary parenchyma cells in sucrose starved lupine embryo axes, whereas cytoplasm along with organelles were forced to a periphery of the cells. We suggest that the autolysis of cytoplasmic proteins contributes to the accumulation of the vacuoles and this suggestion is consistent with the results of the characterisation of protein content. The level of cytosolic proteins was reduced by 50% and the activity of cytosolic marker enzyme, PEP carboxylase, was reduced by 46% in starved embryos as compared to control. The mitochondria from starved tissues were not degraded. The level of mitochondrial proteins was reduced by only 10% and the activity of mitochondrial NAD-isocitrate dehydrogenase decreased by 8% as a result of starvation. As demonstrated by the results of Percoll density gradient centrifugation, sucrose starvation caused an increase of 49% in many of the higher density mitochondria fractions, whereas many of the lower density mitochondria fractions were decreased by 33%. The samples of mitochondria from starved embryo axes were determined to have higher respiration activity in the presence of glutamate and malate as compared to control samples. EPR-based analyses of free radicals showed the presence of free radicals with a signal at g = 2.0060 in embryo axes. The level of the radical was two times higher in sucrose-starved embryo axes than in control (the level of this radical increased in senescing plant tissues as well). The results of EPR based quantitation of Mn2+ ions revealed that the level was a few times higher in starved material than in control. Starved embryo axes, however, do possess a number of adaptive mechanisms protecting them from oxidative damage. Densitometric analyses of gels revealed an increase in the activity of SOD in sugar-starved embryos, whereas CAT and POX activities were lower in axes grown without sucrose as compared to control. Superoxide dismutase, catalase and peroxidase zymogram analyses showed that synthesis of new isoforms was not induced by sugar starvation. An accumulation of phytoferritin was found in plastids of sucrose starved embryos. These results are discussed in relation to the metabolic changes observed in senescing plant tissues. PMID- 12749090 TI - [Counseling--an independent treatment field between old misunderstandings and new positions]. AB - The german term "Beratung" (Counselling) with it's widespread meaning has led to misunderstandings between different professions. Therefore Counselling is defined to be an autonomous area within therapeutic, social, and psychosocial work. The still common perspective which views counselling as a "small" therapy approach derived from therapeutic theory and practice is objected. Besides the international counselling-debates counselling has it's own tradition, theories, concepts, and practices in german discuss too. A difference between counselling and therapy is developed: While therapy is linked to discourses on healing, counselling is linked to discourses on help and support. The ways of providing support and help are changing within a changing society. Besides a still therapeutic oriented view on counselling, and an information oriented perspective a network perspective on counselling-theory, and concepts is proposed to be a frame for coming debates. PMID- 12749091 TI - [Resource oriented counseling--conceptual and methodological considerations]. AB - In the last years resources have become one of the most discussed topics, nearly a magic word, in psychotherapy and in psychosocial practice. The orientation towards human strength and potentials can be considered as a central dimension within the reflections about an independent psychosocial understand of counselling apart the field of psychotherapy. The point of view connected therewith is not only fixed on personal resources as it mostly the case within the methods of psychotherapy but also on the interaction between the individual strength and abilities and the resources regarding the context which is mainly arranged by the social network. In order to realize this interactive view counselling makes use of different methods and strategies according to schools of psychotherapy, especially narrative psychology and community psychology. The perspective of empowerment gives direction of the development of an integrative concept. PMID- 12749092 TI - [Contract oriented help in institutional child rearing and family counseling]. AB - The present paper discusses the concept of Developing Clinical Contracts as a useful means to reflect professional psychosocial help focussing especially on participial forms of help. Asking for help is seen as expression of client expertness. Goals and perspectives of change are based on this client expertness. Contextual frames, premises, and proceedings will be outlined and discussed. Developing Clinical Contracts will be characterised as an effective realisation of concerted action, participially designed to produce desired change fitting into the clients' everyday life. The general attitude could be summarised as some kind of intrepid respecting. PMID- 12749093 TI - [Psychological school counseling]. AB - The work of school-counsellors is widespread, because they work with individuals having problems in school as well as with the school as system. Diagnoses and dyadic counselling on one hand, and organizational counselling on the other hand are landmarks of their profession. Within a case study this counselling approach is developed, showing a problemsolving process within the school-system and family-system. Working within these contexts, linking them and using solution oriented an resource-oriented methods is presented as a primary goal of school counselling. PMID- 12749094 TI - [Cultural approach in nursing care of North African Muslims needing palliative care]. AB - Working in a multi-cultural medical establishment, concerning both patients and nursing staff, calls for deliberations regarding the intercultural as well as palliative approach in nursing care. These deliberations are: 1) intra-personal, in questioning the nursing staff on his/her values, beliefs, convictions, personal culture and identity, as well as opinions regarding health, illness and death. 2) Interpersonal, in recognizing the other person through his/her culture, values, individuality, while taking into account his/her opinions on health, illness and death. The goal of this project is to initiate this intercultural approach by first studying the culture from the perspective of North African Muslims currently hospitalized. It will develop in three steps: The first step to consider consists in training health-care professionals. The second step will lead to the preparation of pedagogical support concerning "nursing care and North African culture". The third step should significantly demonstrate the intercultural approach in clinical nursing procedures. In the future, the project's aim should progress towards a health-care collaboration in which each patient feels implicated regardless of his/her condition, culture, beliefs, education or lifestyle. PMID- 12749095 TI - [For competent cultural care: Purnell's Transcultural Model]. PMID- 12749096 TI - [Elderly patients suffering from dementia in nursing homes: a daily challenge for nurses]. AB - In partnership with the EMS of French-speaking Valais, Switzerland, this study has brought to light certain characteristics in the care of elderly patients suffering from dementia. The interviews were conducted between two professional groups: 19 certified nurses' aides (CNA) and 19 registered nurses (RN). The medical facilities were investigated on the basis of psycho-sociological representations, time, professional security, interactions, the types of services offered, and work satisfaction. The CNAs confuse the depictions of aging and dementia, which corresponds to the belief of the general public. The RNs identify with a professional ideal which, as a consequence, fails to differentiate between the individuals who have been treated. These accounts give a reliable perspective for the two professions, but also serve as a screen for the diversity of situations involving elderly patients suffering from dementia. Concerning the time needed for the supplementary workload, the CNAs and RNs favor its efficiency, and the time needed is justified by the end results. The nurses' aides go about their work essentially on the basis of the tasks and services required by the institution, which is largely inspired by the hospital model. Confronted by the unconventional behavior of the residents suffering from dementia, the risk of assault results in deliberations for CNAs, whereas the unpredictability of the situations is fundamental for RNs. The lack of security measures is equally due to the shortcomings of the medical profession regarding dementia and the specific types of care required. These interactions, while regarded as essential by both CNAs and RNs, are not considered therapeutic, despite the fact that communication is a primary function! Both groups share the types of care offered. Ils limitations are chiefly in the area of satisfaction regarding psychological needs. This health-care project is better able to answer the needs of institutionalized patients rather than in providing individual attention. CNAs and RNs are said to be satisfied by their work with patients suffering from dementia with regards to its utility and professional ideal. In conclusion, the particulars of health care are first of all relative to the context (structure, make-up of personnel, type of residents), relying to a lesser degree on the kind of training obtained by the health care professionals. PMID- 12749097 TI - [Representations of breastfeeding in the patient/nurse midwife relation]. AB - At the crossroads of medical, cultural and psychological fields, breastfeeding can often provoke intense reactions. In the course of my profession as a midwife, I have observed difficulties in the encounters between patients and midwives. In the context of social representations, I wished to explore and compare the representations of breastfeeding of both patients and midwives, which will test the hypothesis that the incongruity in patient/midwife relationships derives from a difference in their respective representations of breastfeeding. In order to demonstrate this, I conducted partially structured interviews with five hospitalized patients three days post natal, and five midwives from the Geneva maternity ward. The qualitative thematic analysis of the discussions has shown large differences in comprehension between the patients and the midwives, notably in the subject of experience, the influence of contextual factors, emotions, and normative ideas. We can concur that, from the beginning, this hypothesis has in large part been validated. Propositions were formulated, and are the road to improvement. Even after childbirth, breastfeeding, apart from nourishing a child, not only serves as a catalyst for emotions, but also for tensions, conflicts, as well as cultural paradoxes, both domestic and cognitive. PMID- 12749098 TI - [Validation of a triage scale: first step in patient admission and in emergency service models]. AB - INTRODUCTION: At present, most emergency services handle the multitude of various demands in the same unity of place and by the same team of nurses aides, with direct consequences on the waiting time and in the handling of problems of varying degrees of importance. Our service examines other administrative models based on a triage of time and of orientation. METHODS AND RESULTS: In a prospective study on 679 patients, we have validated a triage tool inspired from the ICEM model (International Cooperation of Emergency Medicine) allowing patients to receive, while they wait, information and training, based on the resources provided, in order to deal with their particular medical problem. CONCLUSION: The validation of this tool was carried out in terms of its utilization as well as its reliability. It appears that, with the type of triage offered, there is a theoretical reserve of waiting time for the patients in which the urgency is relative, and which could be better used in the handling of more vital cases. PMID- 12749099 TI - [Political problems in nursing]. PMID- 12749100 TI - [Nursing is also the issue]. PMID- 12749101 TI - [How can health remain affordable? (interview by Urs Luthi)]. PMID- 12749102 TI - [Human needs are the primary concern]. PMID- 12749103 TI - [Helping acutely confused patients get back on track]. PMID- 12749105 TI - [Night watch]. PMID- 12749104 TI - [Exceptional conditions for patients and nurses]. PMID- 12749106 TI - [Searching dialogue for wishes]. PMID- 12749107 TI - [For a true system of health]. PMID- 12749108 TI - [Toward strong political engagement of ASI?]. PMID- 12749109 TI - ["What shame to find myself on the ground"!]. PMID- 12749111 TI - [Portrait of a profession to aid travelers]. PMID- 12749110 TI - [The school nurse confronting new challenges once again]. PMID- 12749113 TI - [The end of a particular case]. PMID- 12749112 TI - [Payment of costs for indigent dental care patients]. PMID- 12749114 TI - [Learning: an adventure without end]. PMID- 12749118 TI - Criteria for determining disability in infants and children: failure to thrive. PMID- 12749120 TI - [Andrology in the 21st century: keeping pace with the time]. AB - This editorial briefly reviewed the history of andrology, its rapid development in recent years, the conception of male health, international andrology studies, the achievements of Chinese andrologists and the existing problems. It is highly necessary for Chinese andrology community to strengthen the international academic communication, join the International Society of Andrology and attend its activities actively. PMID- 12749119 TI - Criteria for determining disability in infants and children: short stature. PMID- 12749121 TI - [Changes of 5 alpha-reductase type II activity in sexual gland of diabetic male rats]. AB - OBJECTIVES: To study the changes of sexual gland 5 alpha-reductase type II activity in pubertal and adult rats with diabetes. METHODS: We selected 40 and 90 days old male Wistar rats as pubertal and adult animal model respectively, 30 rats in each group. The rats were randomly divided into three groups: control group (C), diabetic group (D) and diabetes with insulin replacement group (ID). The activity of 5 alpha-reductase type II was measured with thin layer chromatography in the epididymis, prostate and testis. RESULTS: 1. In all sexual glands of pubertal rats, the activity of 5 alpha-reductase type II in D group is significantly lower than that in C and ID groups. 2. In all sexual glands of adult rats. there is no difference in the activity of 5 alpha-reductase type II among these groups. CONCLUSIONS: The activity of 5 alpha-reductase type II is likely to be influenced by metabolic environment, hormonal levels and local specific factors in pubertal rats, but it is relatively stable in adult rats. PMID- 12749122 TI - [Application of two-dimensional electrophoresis and mass spectrometry in the research of testicular proteins]. AB - OBJECTIVES: To evaluate the application of two-dimensional electrophoresis and mass spectrometry in the research of proteins expressed in testis. METHODS: Protein from adult ICR mouse testes was extracted by two-dimensional electrophoresis. Two protein spots were cut from the gel, then the proteins were digested in-gel by enzyme and the generated peptides were measured by matrix assisted laser desorption/ionization-time of flight mass spectrometry. The proteins were identified through database searching. RESULTS: Two spots in Commasie Brilliant Blue-stained gel were identified as serum albumin and protein disulfide isomerase by database searching. CONCLUSIONS: This rapid high resolution and efficient method is a very powerful way to analyze testicular proteins. PMID- 12749123 TI - [Detection and clinical significance of phospholipase A2 in semen of male infertile patients]. AB - OBJECTIVES: To explore the clinical application of anti-human seminal plasma phospholipase A2 (PLA2) monoclonal antibody (McAb) for male infertility. METHODS: Enzyme-linked immunoabsorbent assay (ELISA), immunocytochemistry(ICC), as well as flow cytometry (FCM) analysis were established using two strains anti-human seminal plasma PLA2 McAb prepared by our laboratory to detect the PLA2 content in human seminal plasma and the anterior head region of spermatozoa, respectively. Then the PLA2 content in male infertile patients were compared with that in normal control with fertility. The seminal routine analysis was performed by computer-assisted semen analysis (CASA). RESULTS: The PLA2 content of infertile groups were (31.13 +/- 14.49) ng/ml in azoospermic patients, (17.71 +/- 12.45) ng/ml in oligospermic patients and (16.46 +/- 11.31) ng/ml in patients with normal sperm density, which were all higher than that of normal controls [(8.09 +/- 3.15) ng/ml, P < 0.01]; There was significantly negative correlation between PLA2 content in seminal plasma and sperm density(r = -0.602, P < 0.05), while there was insignificant correlation between PLA2 and sperm motility or percentage of motility. The PLA2 content in the anterior head region of spermatozoon of male infertile groups was significantly lower than that of normal controls by ICC and FCM(P < 0.01). CONCLUSIONS: PLA2 in human seminal plasma is closely related to male fertility, and the PLA2 deficiency in the head of spermatozoa may be one of the reasons causing male infertility. The methods detecting PLA2 content in seminal plasma and the head of spermatozoa can provide powerful evidences for exploring the mechanism of male infertility. PMID- 12749124 TI - [Modified cystectomy with preservation of erectile and ejaculatory functions in men with nonmalignant bladder disease]. AB - OBJECTIVES: To evaluate modified cystectomy with preservation of erectile and ejaculatory functions in men with nonmalignant bladder disease. METHODS: Seven cases with average age of 27 years presented with bladder disease necessitating cystectomy, including 2 cases of tuberculous contractile bladder, 1 case of extensive polypoid cystitis glandularis, 4 cases of late stage of neurogenic bladder. All patients wished to maintain erectile and ejaculatory functions after the operation. We performed a modified simple cystectomy with preservation of the vasa deferens, seminal vesicles, prostate and neurovascular bundles, as well as construction of an Indiana pouch or ileal neobladder. RESULTS: Average operative time was 5 h 45 min without perioperative complications in this group. Follow-up ranged from 9 to 60 months. Erectile and ejaculatory functions were normal in all cases. All patients remained completely continent and no dysuria in neobladder, and there was no difficulty in inserting catheter to empty pouch. Upper urinary tract was in good condition 3 and 24 months after operation. CONCLUSIONS: Modified cystectomy with preservation of the vasa deferens, seminal vesicles, prostate and neurovascular bundles is an effective and reliable option for the patients who wish to maintain their fertility and erectile function after surgery. PMID- 12749125 TI - [Efficacy and safety of PGE1 cream in the treatment of erectile dysfunction]. AB - OBJECTIVES: To evaluate the efficacy and safety of PGE1 cream[Befar, NexMed Pharmaceuticals(Zhongshan) Ltd] on men with ED of various etiologies in China. METHODS: This was a double-blind, randomized (1:1, placebo: PGE1 cream), placebo controlled clinic study of PGE1 cream performed at Peking University Pepole's Hospital for 8 weeks. A total of 42 subjects suffered from erectile dysfunction of psychologic, organic or mixed etiology were screened and randomized, and visited occurred at weeks -4, 0, 2 and 4 weeks covering a 4-week no treatment run in period and a 4-week period of double blind treatment. RESULTS: At week four PGE1 cream was shown to be significantly (P < 0.01) effective over placebo in the sexual function endpoints analyses. The primary efficacy variables (Questions 3 and 4 from IIEF) revealed a statistically significant (P < 0.01) improvement over placebo along with a clinical efficacy change score with an effective rate of 63.16% on PGE1 cream vs 9.52% on placebo. The secondary efficacy variables supported the conclusion of the primary efficacy (assessing the proportion of successful attempts at sexual intercourse 68.42% on PGE1 cream vs 19.05% on placebo), and the global assessment question (treatment had improved their erections, 73.68% on PGE1 cream vs 19.05% on placebo). PGE1 cream was well tolerated when given prn. Subjects in the study had a low discontinuation rate (4.76%), only one subject (2.38%) discontinued due to adverse events. The incidence of adverse events was higher for PGE1 cream (30.00%) than for placebo (4.76%). The common adverse events were mild pain of penis and urethra. CONCLUSIONS: PGE1 cream is an effective, safe and well-tolerated treatment in subjects with erectile dysfunction of organic, psychologic or mixed etiology. PMID- 12749126 TI - [Evaluation for clinical staging parameters of prostate neoplasm]. AB - OBJECTIVES: To evaluate significance of clinical parameters in prostate cancer staging. METHODS: One hundred and twelve patients of prostate cancer were diagnosed by transrectal ultrasound-guided prostate needle biopsies. These cases were staged by pathologic diagnosis, MRI and bone scan. Clinical significance of serum PSA, Gleason score of biopsy, percentage of positive biopsy cores in prostate cancer staging were evaluated. RESULTS: Of 112 patients, 30.4% (34/112) underwent radical retropubic prostatectomy. The serum PSA, Gleason score of biopsy and percentage of positive biopsy cores, were significant correlation with staging prostate cancer (r = 0.698, r = 0.674, r = 0.671, P < 0.001), and no significant difference between staging B and staging C (chi 2 = 2.675, P = 0.096; chi 2 = 0.704, P = 0.401). PSA in patients with stage D had significant difference with others (chi 2 = 5.135, P = 0.023; chi 2 = 4.593, P = 0.032). The sensitivity, specificity and accuracy of PSA were 76.7%, 50.0% and 71.4% respectively. Those of Gleason score and percentage of positive biopsy cores were 83.3%, 77.3%, 82.1% and 77.8%, 54.5%, 73.2% respectively. CONCLUSIONS: The serum PSA, Gleason score of biopsy and percentage of positive biopsy cores had clinical significance in the staging of prostate cancer. Gleason score of biopsy in staging was more accurate than that of the other two parameters and the serum PSA can better predict prostate cancer metastasis. PMID- 12749127 TI - [Measurement of the reactive oxygen species and cytokines in the seminal plasma of leukocytospermic patients]. AB - OBJECTIVES: To detect the levels of reactive oxygen species (ROS), superoxide dismutase(SOD) and interleukin 8(IL-8) in seminal plasma of infertile patients, and evaluate the possible relationship between those levels. METHODS: Semen was collected from normal donors (15 cases), infertile men without infection (16 cases), and infertile men with infection (leukocytospermia, 11 cases). The routine analysis of semen was accomplished, and then the levels of IL-8, malondialdehyde (MDA), SOD, and white blood cell (WBC) were examined. The correlative analysis between the level of ROS and other parameters in these populations was made. RESULTS: In leukocytospermic group, the levels of MDA, WBC, and IL-8 were higher than those in the other two groups (P < 0.001). Significantly positive correlation was observed between IL-8 and MDA (r = 0.852, P < 0.001) and between the levels of IL-8 and WBC. CONCLUSIONS: These findings suggest that increased oxidative stress in patients with leukocytospermia may cause the increase of IL-8(r = 0.818, P < 0.01). The increased oxidative stress may be due to defect in ROS scavenging system. PMID- 12749128 TI - [Effect of aging on the expression of nitric oxide synthase I and the activity of nitric oxide synthase in the rat penis]. AB - OBJECTIVES: To investigate the effect of aging on the expression of nitric oxide synthase I (NOS I) and the activity of nitric oxide synthase in the rat penis. METHODS: Thirty male rats from three age groups(adult, old and senescent) were investigated: 1. The expressions of NOS I protein and mRNA in the penis were detected by Western blot and RT-PCR, respectively. 2. NOS activity in the penis was detected with ultraviolet spectrophotometry. RESULTS: In rats of the old and the senescent groups, the expression of NOS I protein decreased significantly as compared to that of the adult group. NOS I mRNA expression was well related to its protein expression. NOS activity had no statistical difference between the adult group and the old group, but it reduced significantly in the senescent group as compared to that of the adult group(P < 0.01). CONCLUSIONS: It is maybe one of the main mechanisms of erectile dysfunction in the aging male that the aging causes the decreases of NOS I protein and mRNA expression and NOS activity. PMID- 12749129 TI - [Three-piece inflatable penile prosthesis implantation for the treatment of penile erectile dysfunction]. AB - OBJECTIVES: To introduce the method and experience of three-piece inflatable penile prosthesis for treatment of erectile dysfunction (ED). METHODS: Both AMS700CX/CXM and MENTOR ALPHA I three-piece inflatable prosthesis were used for 48 cases of ED. RESULTS: There were no rejection, mechanical problems, urethral injury and penile skin necrosis after surgery. All patients had satisfaction of sexual life. Only one case had trauma of tunica albuginea (2.08%) and the implantation had to be cancelled. There were 2 cases of post-operative infection (4.17%), 10 cases of prepuce edema(20.8%), 8 cases of penile pain(16.67%). The effective rate was 97.9% and satisfaction rate was almost 100%. CONCLUSIONS: Three-piece inflatable penile prosthesis was more compatible to human physiology. It is natural, easy to hide and use, and it caused little trauma in surgery with high rate of successful implantation and patient satisfaction. It was indicated for all kinds of ED, especially suitable for refractory and drug incurable ED patients. PMID- 12749130 TI - [Cloning and eukaryotic expression of human lipocalin-type prostaglandin D synthase in Pichia Pastoris]. AB - OBJECTIVES: To express human testis Lipocalin-type prostaglandin D synthase in Pichia Pastoris for further research on biological function and clinical applications. METHODS: Human testis L-PGDS gene coding region was amplified from plasmid pGEX-2T/htL-PGDS by PCR with a deletion of the signal peptide sequence. The DNA fragment was inserted into pPIC9 to construct yeast expression plasmid followed by transformation of the yeast GS115 strain with electroporation. The recombinant his-tag protein was induced to express by methanol. RESULTS: The sequence of the amplified DNA fragment was identical to that of human testis L PGDS previously reported. The recombinant protein was found with a molecular mass of 27,000 on SDS-PAGE, which was identical to that of native L-PGDS. CONCLUSIONS: Secretory expression of human L-PGDS was obtained in Pichia Pastoris. PMID- 12749131 TI - [Effects of antihypertensive drugs on erectile function and number of nNOS containing nerve fibers in spontaneously hypertensive rats]. AB - OBJECTIVES: To investigate the effects of antihypertensive drugs on the erectile function and number of nNOS-containing nerve fibers in spontaneously hypertensive rats(SHRs). METHODS: Eighteen male SHRs of six-weeks old were randomized into three groups: Vilsartan intervention group, Aldactone intervention group and control group. Vilsartan [30 mg/(kg.d)] and Aldactone [20 mg/(kg.d)] were administered by gavage to respective groups. After twelve weeks treatment, all SHRs were tested for erectile function and then killed for the detection of nNOS containing nerve fibers by Streptaridin-peroxidase immunohistochemistry techniques (SP method). RESULTS: The erection frequency in Vilsartan group was higher, compared with other two groups(P < 0.05), in which no significant difference was observed (P > 0.05). There was no significant difference in erection rates among these three groups. The number of nNOS-containing nerve fibers in two intervention groups was higher than the control group(P < 0.01), but no significant difference existed between these two groups(P > 0.05). CONCLUSIONS: Vilsartan improves erectile function in SHRs while Aldactone doesn't. The difference can't be explained by the number of nNOS-containing nerve fibers, and it is probably related to the vascular reconstruction induced by angiotensin II receptor antagonists. PMID- 12749132 TI - [Etiology and management of persistent hematospermia: a pilot study]. AB - OBJECTIVES: To study the etiology of persistent hematospermia and to evaluate the efficacy of transrectal ultrasonography (TRUS)-guided transperineal needle aspiration and irrigation for diagnosis and treatment of persistent hematospermia. METHODS: Twelve patients were included in the study, with a mean age of (36.4 +/- 10.8) years old, and a mean duration of the disease of (13.9 +/- 6.4) months. After the expressed prostatic secretion (EPS) by prostatic massage was cultured, patients with recurrent hematospermia received TRUS-guided transperineal needle aspiration for seminal vesicle fluid (SVF), which was sent for bacteriological and cytological examination. If the EPS culture were positive, certain antibiotics according to the drug sensitivity assay were injected into the abnormal seminal vesicle(s) via TRUS-guided transperineal needle puncture. The treatment would be repeated one month later if the patients still had hematospermia. The patients were followed up every three months. RESULTS: Abnormal images were found in left seminal vesicle (SV) in 4 cases, right in 3 cases, bilateral in 2 cases, and no abnormal findings in 3 cases. The abnormal findings included: 7 cases of SV and/or ejaculatory duct dilation, 3 cases of thickening SV wall, 3 cases of calcification or calculi of SV, and 1 case of Mullerian duct cyst. SVF cultures were positive in 7 cases: methicillin resistant Staphylococcus aureus (MRSA) 4 cases, methicillin-resistant coagulase negative Staphylococcus (MRCNS), E. Coli, Proteus mirabilis 1 case, respectively. In five of these 7 cases, bacteriological cultures of SVF and EPS showed the same results. All patients were treated by TRUS-guided transperineal injection of certain antibiotics into SV. Seven cases were injected once, 5 cases twice. The mean follow-up period of 10 patients was (16.7 +/- 5.9) months. Hematospermia disappeared in 6 cases. CONCLUSIONS: SV infection of bacteria, especially infection of the drug resistant strains was one of the main causes of persistent hematospermia. The difficulties in treatment of persistent hematospermia were due to infection of drug resistant bacteria, calcification or calculi of SV, obstruction of ejaculatory duct. TRUS-guided transperineal aspiration of SVF was helpful to the etiologic diagnosis of persistent hematospermia. PMID- 12749133 TI - [The effect of the period of cryopreservative storage on the cryosurvival of human spermatozoa]. AB - OBJECTIVES: To study the effect of cryopreservative period on the cryosurvival of human spermatozoa and find out the optimal recovery time of cryopreservation. METHODS: Eighty-eight semen samples were collected from normal donors and divided randomly into 5 groups according to the period of cryopreservative storage (1 d, 7 d, 30 d, 180 d, 300 d) in liquid nitrogen after being frozen by the programized, three-step freezing method. Fresh and frozen-thawed semen were examined by the routine analysis of semen and then the sperm recovery rate were calculated. RESULTS: There were no significant differences in sperm recovery rate between group I and the others (P > 0.05). The period of cryopreservative storage in liquid nitrogen had no correlation with the cryosurvival of human spermatozoa (r = 0.05, P > 0.05). CONCLUSIONS: It was indicated that freezing-thawing after 24 h would be helpful to the screening of semen donors in batches for donor insemination of human sperm bank. PMID- 12749134 TI - [A powerful tool for study of sperm--soft X-ray microscopy]. AB - Soft X-ray microscopy is a microimaging technique using soft X-ray as illuminative source. It fills the gap between optical and electron microscopy. Soft X-ray microscopes have better resolution than visible microscopes. In comparison with electron microscopes, it can examine thick (up to 10 micrometers for biological samples) and wet specimens in their natural states without being dehydrated, sectioned and stained. In addition, soft X-ray microscopy can map elements and analyze the biological macromolecules such as protein and DNA in the examined samples. In this paper, the advantages of soft X-ray microscopy for biology are briefly described. The applications of soft X-ray microscopy to the analysis of mammal and human sperm are illustrated. PMID- 12749135 TI - [The involvement of endocrine disrupting chemicals in spermatogenic cell apoptosis]. AB - Elimination of spermatogenic cells via apoptosis occurs spontaneously under normal physiologic conditions and is often aggravated after chemical-induced testicular impairment. A great amount of pollutants is released into the environment by modern industry, and many of these substances have been confirmed possessing reproductive toxicity, which can affect the reproduction and development of organism. These chemicals have been categorized to endocrine disrupting chemicals(EDCs). Studying spermatogenic cell apoptosis induced by EDCs will enrich and expand the pathway to identify EDCs, and will put forward new expounding of its mechanism. It has important meaning in the field of reproduction toxicology and male fertility. PMID- 12749136 TI - [Relationship between plasminogen activator, plasminogen activator inhibitor and Sertoli cells]. AB - Plasminogen activator(PA) and plasminogen activator inhibitor(PAI) are involved in many physiological or pathological events. The Sertoli cells, the important elements within the seminiferous epithelium, are thought to play a key role in spermatogenesis. The Sertoli cells secrete PA and PAI. The levels of them are modulated by hormonal and cell-mediated influences. They play a fundamental role in the maintenance of spermatogenesis, sperm motility and fertilization. PMID- 12749138 TI - [The effect of Chinese medicine yiqihuoxuetang on T-lymphocyte subpopulation in peripheral blood of infertile men with antisperm antibodies]. AB - OBJECTIVES: To study whether Chinese Medicine Yiqihuoxuetang(YQHXT) could inhibit antisperm antibodies in infertile men, and to explore the therapeutical mechanism of YQHXT. METHODS: Thirty infertile men with antisperm antibodies took YQHXT continuously for 60 days. Indirect immuno-fluorescence technique (IFT) was used to detect the levels of CD3, CD4, CD8 and CD4/CD8 ratio before and after treatment. RESULTS: CD4 value and CD4/CD8 ratio after treatment were significantly lower than before treatment (P < 0.05); CD8 value became significantly higher(P < 0.05). CONCLUSIONS: The results indicated that YQHXT could inhibit antisperm antibodies by keeping the balance of T-lymphocyte subpopulation in immunoinfertile men. PMID- 12749137 TI - [Significance of neutrophil elastase in diagnosis of male genital tract infection]. AB - Neutrophil Elastase(NE) is a kind of neutral proteinase. Its value in the diagnosis of male genital tract infection has attracted increasing attention in recent years. NE has special diagnostic value in screening the typical genital tract infection caused by Neisseria gonorrhoeae, Chlamydia trachomatis, Ureaplasma urealyticum and Trichomonas, as well as other kinds of genital infection, such as leukocytospermia and periorchitis. NE has been considered as a specific marker of the male genital tract infection. PMID- 12749139 TI - Georgia hospitals prepare for April 14 HIPAA implementation deadline. PMID- 12749140 TI - Formal hearing required to revoke physician's license in Florida anti-kickback case. PMID- 12749141 TI - [Endoscopic parathyroid surgery. The time of controversy has passed]. PMID- 12749142 TI - [A researcher in his garden: Henri Dutrochet (1776-1847), the discoverer of the cell]. PMID- 12749143 TI - [Classification of acute coronary syndromes]. AB - The classification of acute coronary syndromes takes into account new data concerning pathophysiology and management. Acute coronary syndromes have a common substrate, namely rupture or erosion of an atherosclerotic plaque leading to thrombus formation and distal embolization, which is responsible for microinfarction with release of myocardial proteins (enzymes and troponins). In the context of acute chest pain, two different presentations of acute coronary syndromes are distinguished on the basis of electrocardiographic findings, according to wether or not there exist persistent ST-segment elevation. Acute coronary syndromes with permanent ST-elevation generally lead to Q-wave infarction and require urgent reperfusion. Acute coronary syndromes without permanent ST-elevation can be further subdivided into: non-Q-wave infarction, when there is release of markers of myocardial cell death, with a prognosis warranting aggressive medical therapy, early coronary angiography and revascularization whenever possible; unstable angina, in the absence of release of markers of cell death, with a more favourable prognosis allowing ambulatory management. PMID- 12749144 TI - [Acute coronary thrombosis]. AB - Atherosclerosis is an inflammatory disease of the arterial wall. Ischaemic manifestations of atherosclerosis are mainly due to thrombus formation upon a superficially eroded (denudation of luminal endothelium, 40% of cases) or deeply ruptured (fibrous cap rupture, 60% of cases) plaques. Recent studies have unraveled potentially critical roles for both inflammatory and apoptotic processes in plaque unstability leading to thrombus formation. Pro-inflammatory mediators have been particularly implicated in the loss of smooth muscle cells and the promotion of collagen degradation that are responsible for fibrous cap rupture, whereas apoptosis has been identified as one of the major determinants of plaque thrombogenicity. PMID- 12749145 TI - [Epidemiology of acute coronary syndrome]. AB - As for myocardial infarction, the epidemiology of acute coronary syndromes is very different according to whether it concerns hospitalized patients only or acute coronary syndromes as a whole: many patients die before any medical intervention. The epidemiology of acute coronary syndromes is not well known since we have almost no specific data regarding unstable angina. The annual incidence of acute coronary syndromes in France is greater than 280 per 100,000 men and 60 per 100,000 women. The 1-month lethality rate is about 50%, increasing sharply with age. PMID- 12749147 TI - [Clinical aspects and risk stratification of acute coronary syndromes without ST segment elevation]. AB - The clinical classification of acute coronary syndromes without ST-segment elevation has long been a topic of debate. Braunwald's classification, however, is now widely adopted and permits to recognize the most severe forms of acute coronary syndromes. Risk stratification is essential and is based upon clinical and biological markers. Among them, recent and repeated anginal attacks, ST segment modifications on admission electrocardiogram, and increased markers of myonecrosis (particularly increased troponine levels) are strong predictors of untoward outcome. These variables can be used to construct risk scores, among which the Timi risk score is the most widely used. PMID- 12749146 TI - [Acute coronary syndrome with ST-segment elevation]. AB - Acute coronary syndromes with ST-segment elevation were known as acute myocardial infarction in the recent past. Their understanding and management are of paramount importance to achieve the earliest reperfusion treatment, using pharmacological (fibrinolysis) or mechanical therapeutics (percutaneous transluminal coronary angioplasty). New fibrinolytics and optimisation of antithrombotic regimen in association with coronary stents have dramatically reduced in-hospital mortality. PMID- 12749148 TI - [Medical treatment of acute coronary syndrome without ST-segment elevation]. AB - Unstable angina is the most frequent acute coronary syndrome. Risk stratification to predict coronary morbidity and mortality and the risk of major haemorrhage are the key steps of the medical approach. Combined antithrombotic therapy (including aspirin, clopidogrel, low-molecular weight heparins and, eventually glycoprotein IIb/IIIa receptor antagonists) has led to a substantial reduction of major coronary events with a good tolerance because of the short duration of such aggressive strategy. This combined antithrombotic also allowed to increase the benefit of an early invasive strategy including coronary angiogram with stent percutaneous coronary angioplasty. PMID- 12749149 TI - [Myocardial revascularization in acute coronary syndrome without ST-segment elevation]. AB - Currently, an invasive strategy is highly recommended in the management of patients with an acute coronary syndrome without persistent ST segment elevation. This early invasive strategy using angiocoronarography allows the identification of the culprit lesion which can be very frequently treated by stent implantation. The use of glycoprotein IIb/IIIa receptor inhibition is recommended especially for high risk patients, not only for the early benefit during medical treatment but also for the additional protection during percutaneous coronary intervention. Patients with left main or three-vessel disease, especially those with associated left ventricular dysfunction or diabetes, are usually managed with coronary artery bypass graft. PMID- 12749150 TI - [Drug prescription after acute coronary syndrome]. AB - Each year in France, 150,000 to 180,000 new patients are the subject of prescriptions following acute coronary syndrome with or without ST segment elevation. There are two targets of the treatment, 1) atherosclerosis, a diffuse, evolving trouble which, in this situation, is coming out of an unstable phase, and 2) the myocardium, which has often been revascularized and has suffered deterioration of its contractile and electrophysiological characteristics to a greater or lesser extent. Prescriptions, based on proven factors and always centred around hygiene and dietary advice and the use of a combination of statins and aspirin, are adapted to suit the atherosclerotic and myocardial risk assessed for the individual patient. The prescription starts off the secondary preventive phase. It marks the first stage of the follow up, which is inevitable though of variable duration, for a disease which may evolve. It is the first step in the accompaniment of an attentive, informed patient whose confidence has been restored and whom must now avoid falling into the double trap of not taking the treatment sufficiently seriously or of obsessively overreacting. PMID- 12749151 TI - [Advice for students and young physicians]. PMID- 12749152 TI - [Diagnosis of cancer. Signs and paraclinical investigations; stratification; prognosis]. PMID- 12749153 TI - [Intestinal obstructive syndrome]. PMID- 12749154 TI - [Arthritis]. PMID- 12749156 TI - [Acute agitation and delirium]. PMID- 12749155 TI - [Travel in tropical countries. Advice before departure. Diseases upon return: fever, diarrhea]. PMID- 12749157 TI - FDA clarifies importation law as Internet pharmacies proliferate. PMID- 12749158 TI - HRSA reorganization could affect 340B program. PMID- 12749159 TI - GAO suggests Medicare make separate payments for clotting factors, delivery. PMID- 12749160 TI - Medications commonly cause problems for community-dwelling elderly. PMID- 12749161 TI - Smallpox: a review of clinical disease and vaccination. AB - The clinical course of smallpox infection and the current and future roles of vaccination and strategies for controlling smallpox outbreaks are reviewed. Close personal contact is required for transmission of variola, the DNA virus that causes smallpox. Following an incubation period, infected persons have prodromal symptoms that include high fever, back pain, malaise, and prostration. The eruptive stage is characterized by maculopapular rash that progresses to papules, then vesicles, and then pustules and scab lesions. The mortality rate for smallpox is approximately 30%. Patients having a fever and rash may be confused with having chickenpox. The most effective method for preventing smallpox epidemic progression is vaccination. Until recently, only 15 million doses of smallpox vaccine--manufactured 20 years ago--were available in the United States. The vaccine is a live vaccinia virus preparation administered by scarification with a bifurcated needle. The immune response is protective against orthopoxviruses, including variola. Vaccination is associated with moderate to severe complications, such as generalized vaccinia, eczema vaccinatum, progressive vaccinia, and postvaccinial encephalitis. Efforts for vaccine production are now focused on a live cell-culture-derived vaccinia virus vaccine. Although smallpox was eradicated in 1980, it remains a potential agent for bioterrorism. As a category A biological weapon, its potential to devastate populations causes concern among those in the public health community who have been actively developing plants to deal with smallpox and other potential agents of biological warfare. The only proven effective strategy against smallpox is vaccination. PMID- 12749162 TI - Effect of an educational program on the treatment of RSV lower-respiratory-tract infection. AB - The effectiveness and outcomes of an educational program to decrease ribavirin and antimicrobial prescribing rates and associated costs for patients with respiratory syncytial virus (RSV) lower-respiratory-tract infection are described. An educational program on the appropriate treatment for RSV infections was conducted for attending physicians and medical residents with multiple methods and forums during the 1994-95 RSV season. A retrospective chart review of 2396 patients admitted to a pediatric teaching hospital from July 1, 1991, through June 30, 1998, was conducted to measure the frequencies of ribavirin and antimicrobial prescribing in infants and young children hospitalized with RSV lower-respiratory-tract infection. The results before and after the educational program were compared. Ribavirin was prescribed for 37.9% of patients before the program, and only 9.0% received it afterward (p < 0.001). Before the program, 24.8% of patients with no risk factors received ribavirin compared with 1.6% of patients after the program (p < 0.001). However, more patients with three or more risk factors for morbidity and mortality received ribavirin before the program than afterward (97.8% versus 39.2%, respectively). A broad-spectrum i.v. antimicrobial was prescribed for 85.6% of patients before the program while 60.6% received one afterward (p < 0.001). The cost savings for ribavirin and antimicrobials during the three-year period after the program were $1,235,484 and $34,839, respectively. Hospital length of stay decreased from 5.6 to 5.1 days (p < 0.001). No readmissions occurred during the study period. A multifaceted educational intervention program may have been somewhat effective in modifying physician's prescribing habits for the treatment of RSV lower-respiratory-tract infection. PMID- 12749163 TI - Practical guide to bar coding for patient medication safety. AB - Bar coding for the medication administration step of the drug-use process is discussed. FDA will propose a rule in 2003 that would require bar-code labels on all human drugs and biologicals. Even with an FDA mandate, manufacturer procrastination and possible shifts in product availability are likely to slow progress. Such delays should not preclude health systems from adopting bar-code enabled point-of-care (BPOC) systems to achieve gains in patient safety. Bar-code technology is a replacement for traditional keyboard data entry. The elements of bar coding are content, which determines the meaning; data format, which refers to the embedded data and symbology, which describes the "font" in which the machine-readable code is written. For a BPOC system to deliver an acceptable level of patient protection, the hospital must first establish reliable processes for a patient identification band, caregiver badge, and medication bar coding. Medications can have either drug-specific or patient-specific bar codes. Both varieties result in the desired code that supports patient's five rights of drug administration. When medications are not available from the manufacturer in immediate-container bar-coded packaging, other means of applying the bar code must be devised, including the use of repackaging equipment, overwrapping, manual bar coding, and outsourcing. Virtually all medications should be bar coded, the bar code on the label should be easily readable, and appropriate policies, procedures, and checks should be in place. Bar coding has the potential to be not only cost-effective but to produce a return on investment. By bar coding patient identification tags, caregiver badges, and immediate-container medications, health systems can substantially increase patient safety during medication administration. PMID- 12749164 TI - Not too perfect: hard lessons and small victories in patient safety. PMID- 12749165 TI - Role of angiotensin-converting-enzyme inhibitors in stroke prevention. PMID- 12749166 TI - John A. Williams. 76th President of APS. Presidential address. PMID- 12749167 TI - Visas for visiting scientists and students: current situation. PMID- 12749168 TI - UCSD dog labs do have value. PMID- 12749169 TI - President Bush proposes FY 2004 spending plan. PMID- 12749170 TI - [The role apoptosis in the extent of endometriosis and its clinical symptomatology]. AB - OBJECTIVE: To evaluate the presence and intensity of apoptosis in lesions of peritoneal endometriosis. To consider the role of different intensity of apoptosis in the progression of the disease and in the manifestation of clinical symptoms. DESIGN: Prospective study. SETTING: Department of Obstetrics and Gynecology and Department of Histology and Embryology, 1st Medical Faculty, Charles University and General Faculty Hospital, Prague. METHODS: Lesions of peritoneal endometriosis were excised at laparoscopy in women with 1st to 3rd stage of the disease. Specimens were fixed in the Karnowski solution. The presence of apoptosis was assessed immunohistochemically. RESULTS: Biopsies of peritoneal endometriosis were consecutively taken in 48 women. Altogether, 29 patients were symptomatic, and 19 did not complain of any symptoms. The average duration of symptoms was 16.6 months. The presence of apoptosis was detected in 11 out of 35 evaluated specimens. In comparing the groups with and without apoptosis, no differences were found in the stage of the disease, in the proportion of asymptomatic patients, and in the manifestation of different subjective complaints. CONCLUSIONS: The study confirmed a frequent presence of apoptosis in biopsies of peritoneal endometriosis. However, there were no relationships found between the presence of apoptosis and the stage of the disease or manifestation or character of subjective complaints. PMID- 12749171 TI - [Long-term follow-up after complete treatment of peritoneal endometriosis with the CO2 laser]. AB - OBJECTIVE: To evaluate the effect of laparoscopic CO2 laser ablation of peritoneal endometriosis in the treatment of pelvic pain with a long-term follow up. To differentiate the effect of surgery on different types of pelvic pain. DESIGN: Prospective observational study. SETTING: Department of Obstetrics and Gynecology, 1st Medical Faculty, Charles University and General Faculty Hospital, Prague. METHODS: Patients with 1st to 3rd stage endometriosis, with manifestation of pelvic pain, and with complete excision of peritoneal endometriosis lesions, were included in the study. All visible lesions were vaporized by CO2 laser following adhesiolysis and complete visualization of the pelvis. After the procedure, patients were followed up at 6-month intervals. The severity of pelipathia, dyspareunia, dysmenorrhea, pain during micturition, and pain during defecation were monitored using a visual analog score of 10 points. RESULTS: A total of 31 patients were included in the study. After 6, 12, and 18 months after surgery, the recurrence of pelvic pain was found in 12 (39%), 15 (48%), and 19 (61%) patients, respectively. Improvement or disappearance of complaints was documented 18 months after the surgery in 11 cases of dysmenorrhea (50%), 9 cases of dyspareunia (50%), 14 cases of pelipathia (58%), 12 cases of pain during micturition (71%), and in 14 cases of pain during defecation (87.5%). The proportion of recurrences increases with the length of the interval after the procedure, mainly in dysmenorrhea and dyspareunia. CONCLUSIONS: The effect (improvement or disappearance of pelvic pain) of a complete CO2 laser ablation of peritoneal endometriosis continues 18 months after the surgery in about 40% of patients. A graduated increase in the number of recurrences is apparent during follow-up, most significantly in dysmenorrhea and dyspareunia. The effect of surgery on different types of pelvic pain varies. A small number of recurrences was found in pain during micturition and pain during defecation, on the other hand, less success was apparent in the treatment of dysmenorrhea. PMID- 12749172 TI - [Importance of regular screening for genital chlamydiosis in adolescents]. AB - OBJECTIVE: Determination of prevalence of genital chlamydiosis in similar group of adolescents as in that examined two years ago. Comparison of the results with the previous ones to confirm positive impact of a repeated screening on drop of prevalence of the infection. Another objective is to make the epidemiological data on genital chlamydiosis in the countries of the "Eastern Block" more accurate, and at the same time to assess the most suitable screening method. DESIGN: Prospective epidemiological study aimed at finding whether the repeated screening examinations of genital chlamydiosis in adolescents will result in decreased incidence of the infection. SETTING: Veterinary Research Institute, Department of Clinical Microbiology of the Faculty Hospital, Regional Hygienic Station, Department of Gynecology and Obstetrics, Medical Faculty, Masaryk University, Brno. METHODS: The groups under testing comprised male and female students of two secondary medical schools and one private grammar school older than 18 (n = 232, of which 33 were boys and 199 girls). Direct detection of C. trachomatis was carried out from the urinary sediment of the sample of morning urine using ligase reaction, polymerase chain reaction, ELISA and direct immunofluorescence. RESULTS: Identical positive result of C. trachomatis detection by four methods was obtained 4 times, i.e. 1.7%. The result of LCR and PCR were positive identically (4 times), ELISA and IF were, besides the above results, positive in other 1 resp. 5 cases. CONCLUSION: Screening examinations of genital chlamydiosis were performed in adolescents in the town of Brno. Morning samples of urine from 232 students of secondary schools were examined on C. trachomatis using the following methods: LCR, PCR, ELISA, and IF. The LCR and PCR were positive 4 times, ELISA 5 times, IF 9 times. Positive results by LCR and PCR were identical. ELISA and IF were positive in other cases. Noninvasive methods are suitable for screening of latent genital chlamydiosis and for estimation of its prevalence. The authors of the study recommend the ligase chain reaction (LCR) as a most suitable screening method. PMID- 12749174 TI - [Celiac disease and fertility disorders in women]. AB - OBJECTIVE: To determine incidence of subclinical forms of celiac disease in women with decreased fertility. DESIGN: Screening test. SETTING: Department of Gynecology and Obstetrics, University Hospital, Palacky University, Olomouc. METHODS: 137 patients with fertility problems were included to the study. There were divided into two groups, patients with infertility and patients with repeated pregnancy loss. Screening test for celiac disease, serum level of antibodies tissue transglutaminase (tTGA) were performed in all of them. Positive test was confirmed by serological level of anti endomysium antibodies (EmA) and final diagnosis of silent celiac disease was done by enterobiopsy. RESULTS: Celiac disease was found in two infertile patients (1.67%). In one patient the silent form was diagnosed by enterobiopsy. The second disease was latent form. In patients with repeated pregnancy loss we did not observe positive screening test. CONCLUSION: We confirmed higher incidence of celiac disease in women with impaired fertility. PMID- 12749173 TI - [Routine monitoring of congenital defects in children in the Czech Republic. History and present status]. AB - OBJECTIVE: Analysis of quarterly incidences of selected types of birth defects in the Czech Republic in 1994-2000. SETTING: Institute for the Care of Mother and Child, Prague; Department of Medical Genetics, Thomayer Hospital, Prague and Institute of Health Information and Statistics, Prague. TYPE OF STUDY: Retrospective analysis of quarterly data from registers of birth defects in the Czech Republic. METHODS: Processing and analysis of incidences of selected birth defects in the Czech Republic in relative numbers (per 10,000 liveborn infants). The ratio of observed/expected values was calculated. RESULTS: During the period between Jan. 1 1994 and Dec. 31 2000 a total of 654 695 liveborn infants were born (for 2000 preliminary data). This number included 18,008 infants with one or more birth defects. The mean incidence of all cases during the observation period was 262.99 per 10,000 liveborn infants. In the analysis the authors discuss in detail also the frequency and its changes of eleven selected types of birth defects. CONCLUSION: Continuous registration and analysis of the incidence of inborn defects are essential as the basis for evaluation of the influence of the mentioned factors on the incidence of inborn defects. PMID- 12749175 TI - [Occurrence of bleeding in women using combined hormonal contraceptives (ethinylestradiol 35 micrograms/norgestimate 250 micrograms in relation to regularity of administration and cycle start day]. AB - OBJECTIVE: To assess the frequency of irregular intermenstrual bleeding in combined oral contraceptive (ethinylestradiol 35 micrograms/norgestimate 250 micrograms, COC) users and the influence of regularity of pill use on this frequency; to assess the occurrence of withdrawal bleeding during weekends in women using the COC from the first Sunday in the cycle (Sunday start method). DESIGN: Prospective, open, non-comparative, multicenter study in 27 centers. METHODS: The first day of the pill use, occurrence of intermenstrual and withdrawal bleeding and regularity of use were assessed by means of patient's bleeding diary. Body weight, blood pressure and side effects were monitored before the oral contraceptive use and after the third cycle. RESULTS: 358 (94%) of 382 women completed the study. Frequency of intermenstrual bleeding was generally low (6.7%, 5.0% and 5.0% in the first, second and third cycle) and highly influenced by regularity of pill use (2.6%, 0% a 1% in regular users versus 30.2%, 32.1% a 24.2% in irregular users). 28%, 40% and 47% of Sunday start users achieved bleeding-free weekends after the first, second and third cycle. Body weight and blood pressure did not change during the study. CONCLUSION: Frequency of intermenstrual (breakthrough) bleeding during the first three months of COC use is highly influenced by regularity of use. In regular users of monophasic COC containing ethinylestradiol 35 micrograms/norgestimate 250 micrograms the frequency of intermenstrual bleeding is bellow 2.6%. In Sunday start users the proportion of women with bleeding-free weekends (i.e. weekends without the occurrence of withdrawal bleeding) increases to 47% after the third cycle. PMID- 12749176 TI - [Preimplantation genetic diagnosis (PGD) of chromosomal aberrations using the fluorescence in situ hybridization method (FISH)--introduction to problems, sampling methods and examination techniques]. AB - OBJECTIVE: Presentation of preimplantation genetic diagnosis and the set of laboratory processes like aspiration, preparation and evaluation of polar bodies, sperm cells and blastomeres using FISH method (fluorescent in situ hybridization) in ART. DESIGN: Review. SETTING: Sanatorium REPROMEDA, Brno, Veterinary Research Institute, Brno. METHODS: Overview of published data and own clinical experience with the cell aspiration methods, evaluated sample preparation and the proper chromosomes visualisation using FISH method. CONCLUSION: The review brings an overview of conditions and methods including sample obtaining, FISH analysis preparation and implementation, processed during PGD. PMID- 12749177 TI - [Effectiveness and safety of the tension-free vaginal tape (TVT) operation during the learning period]. AB - OBJECTIVE: To evaluate the safety and efficacy of the tension-free vaginal tape procedure in the treatment of female stress urinary incontinence during the learning phase. DESIGN: Retrospective clinical trial. SETTING: Obstetrics and Gynecology Department, 2nd Medical Faculty, Charles University and Faculty Hospital Motol, Prague. METHODS: A group of first 20 patients with stress urinary incontinence who underwent tension-free vaginal tape procedure was studied. Surgical procedure was performed according to Ulmsten's technique under the spinal anaesthesia. All patients were operated on by one surgeon and none had undergone any previous anti-incontinence surgery. Preoperative evaluation consisted of urodynamic examination, Q-tip test, stress test, age, parity, body mass index, history of previous pelvic surgery and hormonal status. The cure rate and complications were determined in short-term postoperative follow-up. RESULTS: The mean age was 59.5 (43-74) years, the median parity was 1.55 (1-2), 16 (80%) patients were slightly overweight (BMI 25-30 kg/m2) and others had normal weight (20-25 kg/m2). The cure rate in 6 months follow-up was 95% (19 of 20 patients). The complications occurred in 7 (35%) cases. 5 (25%) patients had mild early postoperative complications (two lower urinary tract infections, one defect healing of vaginal suture, one urge symptomatology, one short-time urinary retention) and 2 (10%) had serious late postoperative complications (urethral diverticulum, retropubic haematoma) which required surgery (diverticulectomy, transabdominal evacuation of the haematoma). All complications were resolved and the patients were 6 month after the procedure free of negative postoperative symptoms. CONCLUSIONS: This study shows that tension-free vaginal tape procedure is an effective and safe minimally invasive surgical procedure in the treatment of stress urinary incontinence in short-term follow-up. The study also suggests that a learning phase of approximately 20 operations performed in a standard manner by one surgeon is required for good results. The operative technique is simple but the TVT procedure should be performed by experienced urogynecologists who are able to resolve intraoperative and postoperative complications. PMID- 12749178 TI - [Para-aortic lymphadenectomy in gynecology. Theoretical principles, basic approach and practical suggestions]. AB - OBJECTIVE: To present a review of para-aortic lymphadenectomy in gynecology. SUBJECTS: Review. SETTING: Department of Obstetrics and Gynecology, Department of Urology, Medical Faculty Hradec Kralove, Charles University, Prague; Bristol Myers Squibb, Department of Oncology, Prague. SUBJECT AND METHOD: Discussion about current evidence from literature and our own experience. CONCLUSION: In the past 20 years, pelvic and paraaortic lymphadenectomy has become part of the standard surgical treatment of ovarian cancer. Overall, about 40% of patients with ovarian cancer will have nodal involvement. Regarding to this real probability is para-aortic lymphadenectomy very important part of surgical treatment in this disease. PMID- 12749179 TI - [Sperm antibodies and immunologic intolerance to metals in infertile couples]. AB - OBJECTIVE: Verification of the hypothesis of a relationship between the presence of antibodies against sperm cells and immunological reactivity to some metals in infertile couples by the MELISA test. SETTING: Department of Obstetrics and Gynecology, Medical Faculty, Charles University and Faculty Hospital, Plzen. METHOD: From 23 female patients and 21 men (a total of 44 subjects treated for infertility) with confirmed serum antibodies against sperm cells the authors isolated lymphocytyes from the peripheral blood stream, divided them into individual cultures and investigated them by the MELISA test using different metal compounds. RESULTS: The outcome of the MELISA test are values of the stimulation index (SI) by means of which the authors investigated the reactivity of the organism to the given metal. Special attention was devoted to compounds of organic and inorganic mercury. The SI values were subsequently compared with different data obtained from a detailed anamnestic questionnaire which was focused specially on contact with metals and on allergic reactions. In the investigated group of patients the authors detected a positive immune reactivity to inorganic mercury, Ag, Al, Fe. In some subjects they found a very high positive immune reactivity to inorganic mercury, Ni, Al, Cd and Ti. The control groups were formed by healthy fertile subjects without antibodies against sperm cells and with physiological SI values. CONCLUSION: The authors did not prove a direct relationship between the intensity of the laboratory reactivity to metals and the presence of antibodies against sperm cells which cause deterioration of fertility. An exogenous load of metals could in case of genetic predisposition be only one of the factors which participate in the formation of antibodies against sperm cells. The investigation proved that its is not essential, contrary to the view of many stomatologists, to eliminate metal compounds completely from dental practice. PMID- 12749180 TI - [Obstetrical aspects of peripartum cardiomyopathy]. AB - OBJECTIVE: The authors demonstrate a case report of the peripartum cardiomyopathy that occurred at a so far healthy woman after the delivery by the caesarean section. SUBJECT: Case report. SETTING: Department of Obstetrics and Gynecology, 1st Medical Faculty, Charles University and General Faculty Hospital, Prague. METHODS: The gravidity of a 28-year old primipara, gemini, was finished by the caesarean section because of cardiologic (benign cardiac extrasystoles) and obstetric (gemini clashing position) indication. After the delivery a heart failure started to develop. The diagnosis was closed as peripartum cardiomyopathy. The clinical state of patient stabilized after initiating adequate therapy, the follow up has been still going on. RESULTS: Peripartum cardiomyopathy is a rare disease that occurs as a left ventricular cardiac failure in pregnant women or in woman in childbed. Etiology of the disease is unknown, the disease management requires collaboration among obstetricians, cardiologists, anesthesiologists and neonatologists. Prognosis is uncertain. PMID- 12749181 TI - [Internal jugular vein thrombosis in a patient with ovarian hyperstimulation syndrome]. AB - OBJECTIVE: To describe a case of right internal jugular vein thrombosis complicating ovarian hyperstimulation syndrome (OHSS). DESIGN: Case report. SETTING: 1st Medical Faculty, Charles University and General Faculty Hospital, Prague. PATIENT: A 27-year-old primiparous woman undergoing in vitro fertilisation (IVF). PMID- 12749182 TI - [Does grapefruit juice increase the bioavailability of orally administered sex steroids?]. AB - OBJECTIVE: To verify if and to which extent the interaction with grapefruit juice can increase bioavailability of orally administered sexual steroids. DESIGN: Pilot pharmacokinetics study. SETTING: Department of Obstetrics and Gynecology and Institute of Pharmacology, Medical Faculty, Palacky University, Olomouc; Department of Nuclear Medicine, University Hospital, Olomouc. METHODS: 2 mg of estradiol valerate and 100 mg of micronized progesterone were given to eight healthy postmenopausal volunteers. Blood samples were collected at time 0, 2, 3, 5 and 24 hours after tablets application. The same trial was repeated a week later but tablets were swallowed with 200 ml of grapefruit juice. Serum levels of estradiol and progesterone were measured by RIA. Results were statistically evaluated using the Wilcoxon's nonparametric paired test. RESULTS: Though grapefruit juice on average slightly increased serum levels of estradiol (E2) and progesterone, this increase reached statistical significance only for the E2 level 24 hours after application of tablets. The mean area under curve (AUC) of estradiol rose significantly to 117%. The even greater increase in the mean AUC of progesterone (to 125%) was not statistically significant because of marked individual variability of response. CONCLUSIONS: Our results suggest that grapefruit juice may increase bioavailability of orally administered estradiol and progesterone. The response varies markedly between individuals. This observation may be of some importance also for users of OC and HRT. PMID- 12749183 TI - [Oocyte donation: ethical and practical questions]. AB - AIM OF THE STUDY: This review will summarize the current view on ethical and practical aspects involved in ovum donation. STUDY DESIGN: Literature review. INSTITUTION: Center of Reproductive Medicine, Zlin; Department of Gynecology and Obstetrics, Palacky University, Olomouc. CONCLUSION: Today there are three possible ways of recruiting ovum donors: 1) volunteer ovum donors, 2) related donors (searched by couples in need), 3) in vitro fertilization patients (egg sharing). PMID- 12749184 TI - [Recommendations for hormone replacement therapy in perimenopause and postmenopause]. PMID- 12749185 TI - [Minimally invasive therapy of benign uterine tumors: strategies and facts]. PMID- 12749186 TI - [An alternative method of treating myomas using uterine artery embolization- considerations after 3 years]. PMID- 12749187 TI - [Hysteroscopy in the therapy of uterine myomas]. PMID- 12749188 TI - [Nurses and geriatrics. Aging and health]. PMID- 12749189 TI - [Simultaneous determination of five penicillins in muscle, liver and kidney from slaughtered animals using liquid chromatography coupled with electrospray ionization tandem mass spectrometry]. AB - A simple and rapid method for the simultaneous determination of five penicillins (ampicillin, penicillin G, penicillin V, oxacillin and cloxacillin) in muscle, liver and kidney tissues using high-performance liquid chromatography electrospray tandem mass spectrometry (LC-ESI-MS/MS) was developed. Mass spectral acquisition was done in the negative ion mode by applying selected reaction monitoring (SRM). The five penicillins were extracted with water, and the extracted solution was cleaned up on a C18 cartridge. Phenethicillin was added as an internal standard, and the extract was diluted with water for injection into the LC-ESI-MS/MS. The recoveries of the five penicillins were in the range of 77.3-99.8% from muscle, liver and kidney fortified at 10-250 ng/g. The detection limits for ampicillin were 6 ng/g in muscle and kidney and 15 ng/g in liver. For penicillin G and penicillin V, the detection limits were 2 ng/g in muscle and kidney and 5 ng/g in liver. For oxacillin and cloxacillin, the detection limits were 4 ng/g in muscle and kidney and 10 ng/g in liver. Twenty-three muscle, fourteen liver and twenty-two kidney samples from the markets were analyzed by this method. No penicillins were detected in any sample. PMID- 12749190 TI - [Effects of rice cleaning and cooking process on the residues of flutolanil, fenobucarb, silafluofen and buprofezin in rice]. AB - We studied the effect of cleaning and cooking on the residues of flutolanil, fenobucarb, silafluofen and buprofezin in rice. The rice had been sprayed in a paddy field in Wakayama city, with 3 kinds of pesticide application protocols: spraying once at the usual concentration of pesticides, repeated spraying (3 times) with the usual concentration of pesticides and spraying once with 3 times the usual concentration of pesticides. The residue levels of pesticide decreased during the rice cleaning process. Silafluofen, which has a higher log Pow value, remained in the hull of the rice. Fenobucarb, which has a lower log Pow value, penetrated inside the rice. The residue concentration of pesticide in polished rice was higher than that in pre-washed rice processed ready for cooking. During the cooking procedure, the reduction of pesticides in polished rice was higher than that in brown rice. PMID- 12749191 TI - A simple and reproducible testing method for dialkyl phthalate migration from polyvinyl chloride products into saliva simulant. AB - We describe a method of mechanical agitation to determine rates of dialkyl phthalate migration from polyvinyl chloride (PVC) products into saliva simulant. The method consists of rotary shaking of a sample with 30 mL of saliva simulant (pH 7.0) at 35 degrees C in a 50 mL glass tube at 300 rpm for 15 min, then measuring the amount of dialkyl phthalate in the saliva simulant by HPLC with a UV detector. The migration rates of diisononyl phthalate (DINP), di-2-ethylhexyl phthalate (DEHP) and di-n-butyl phthalate (DBP) from PVC plates containing about 45% (w/w) plasticizer (molded in our laboratory) were identical. However, the migration rates from molded plates containing 13% (w/w) DBP were almost double those of DINP and DEHP at the same ratios. In addition, the amounts of DINP that migrated in vitro after rotary shaking for 15 min were equivalent to those in vivo determined in saliva from volunteers who chewed plates for 60 min. The migration rates of dialkyl phthalates from 11 commercially available toys ranged from 15.6 to 85.2 micrograms/cm2/h [relative standard deviation (RSD), 3 to 12%]. PMID- 12749192 TI - [Simultaneous analysis of four kinds of emulsifiers in beverages by GC/MS]. AB - An analytical method using GC/MS for the detection of 4 kinds of dietary emulsifiers, glycerin, sucrose, sorbitan and propylene glycol monoesters of fatty acids (GE, SuE, SE, PGE), in beverages was developed. The emulsifiers were extracted from beverages with tetrahydrofuranethyl acetate (6:4) by homogenizing. The extract was cleaned up on a silica gel column and subsequently a C8 cartridge column, followed by acetylation. The derivatives were then detected by GC/MS. Our newly established method enabled to characterize 4 kinds of emulsifiers and also to identify their fatty acids without hydrolysis or de-esterification. When this method was applied to various beverages on the market, many GE and SuE with different fatty acids were detected. These results suggested that several dietary emulsifiers are used as food additives at the same time in beverages on the market. PMID- 12749193 TI - [Determination method of tricresyl phosphate in polyvinyl chloride]. AB - A simple and rapid method using HPLC was developed for the determination of tricresyl phosphate (TCP) in polyvinyl chloride (PVC) articles. A test sample was extracted with acetonitrile at 37 degrees C overnight. The extract solution diluted with an equivalent amount of water was applied to a Sep-Pak C18 cartridge, and TCP was eluted with acetonitrile-water (2:1) mixture. The eluate was analyzed by HPLC with an Inertsil Ph-3 column, using 65% acetonitrile/water as the mobile phase, with UV detection (264 nm). The calibration curve was rectilinear from 0.5 to 100 micrograms/mL. The recoveries of TCP added to various kinds of PVC articles at the level of 1,000 micrograms/g were 84.7-98.6%. The determination limit of TCP was 50 micrograms/g in samples. This method was applied to products including 3.1, 6.6 and 8.8% TCP and the recoveries of TCP were 87.3-91.4%. This method is very simple, and it seems suitable for a regulatory test. PMID- 12749194 TI - [Simulation of migration from internal can coatings]. AB - Migration from can-coatings into retorted canned food simulants (canned oil and water, 121 degrees C, 30 min) was investigated through HPLC with a fluorescence detector and evaporative light scattering detector, and by measurements of residue on evaporation and consumption of potassium permanganate. HPLC analysis revealed that migration into the canned oil was hundreds of times more than that into n-heptane (25 degrees C, 60 min, the official test conditions according to the Japanese Food Sanitation Law), whereas it was similar to the migration into isooctane-butyl acetate mixtures (60 degrees C, 60 min), and that migration into the canned water was several times more than that into water (95 degrees C, 30 min, the official test conditions). Residue on evaporation for the n-heptane extract was several-fold lower than 30 ppm (the official limit), whereas that for the isooctane-butyl acetate mixtures exceeded 30 ppm. Consumption of potassium permanganate for the canned water was 30 times higher than that for the water extract (95 degrees C, 30 min). The official test conditions for can-coatings, in particular the use of n-heptane as an oil simulant, were suggested to lead to substantial underestimation of migration into canned food. PMID- 12749195 TI - [Improved method for determining bisphenol A in polycarbonate products without using dichloromethane]. AB - An improved method for determination of bisphenol A (BPA), phenol (PH), p-tert butylphenol (PTBP) and diphenylcarbonate (DPC) in polycarbonate products was developed without using the hazardous solvent dichloromethane. Polycarbonate samples were ground to powder or cut into small pieces, and 0.5 g was soaked in 5 mL of acetonitrile for 24 hours at 40 degrees C. The test solution was then filtered and subjected to HPLC analysis. The proposed method was evaluated by comparison of the results with those of the standard method for 14 polycarbonate products. Extraction ratios (average values obtained by the present method/average values obtained by the standard method) of BPA, PH, PTBP and DPC were 0.89-1.19, 0.89-1.14, 0.94-1.30 and 1.08-1.11, respectively. While 120 mL/sample of organic solvent is required in the standard method, only 5 mL/sample of acetonitrile was used in our new method. PMID- 12749196 TI - [Discrimination of psychoactive fungi (commonly called "magic mushrooms") based on the DNA sequence of the internal transcribed spacer region]. AB - 'Magic mushrooms' (MMs) are psychoactive fungi containing the hallucinogenic compounds, psilocin (1) and psilocybin (2). Since June 6, 2002, these fungi have been regulated by the Narcotics and Psychotropics Control Law in Japan. Because there are many kinds of MMs and they are sold even as dry powders in local markets, it is very difficult to identify the original species of the MMs by morphological observation. Therefore, we investigated the internal transcribed spacer (ITS) region in the ribosomal RNA gene of MMs obtained in Japanese markets to classify them by a genetic approach. Based on the size and nucleotide sequence of the ITS region amplified by PCR, tested MMs were classified into 6 groups. Furthermore, a comparison of the DNA sequences of the MMs with those of authentic samples or with those found in the databases (GenBank, EMBL and DDBJ) made it possible to identify the species of tested MMs. Analysis by LC revealed that psilocin (1) was contained at the highest level in Panaeolus cyanescens among the MMs, but was absent in the Amanita species. PMID- 12749197 TI - Effects of exogenous polyamines on growth, toxicity, and toxin profile of dinoflagellate Alexandrium minutum. AB - The effect of exogenous polyamines (cadaverine, putrescine, norspermidine, spermidine, and spermine) on the growth, toxicity, and toxin profile of the dinoflagellate Alexandrium minutum T1 was examined. It was found that cadaverine at concentrations of 0.1-2.0 mumol/L enhanced the growth of A. minutum T1. Putrescine and norspermidine at a low level (0.1 mumol/L) also promoted the algal growth. Spermidine depressed the algal growth. However, the cell toxicity levels of A. minutum T1 cultured with or without cadaverine, putrescine, norspermidine, and spermidine were almost the same. The toxic components of A. minutum T1 were GTXs 1-4 only, and GTXs 1 and 4 were predominant (74.6 +/- 7.1%) in all cultures. On the other hand, spermine did not effect the growth of A. minutum T1, though it decreased the cell toxicity and the ratio of GTX 2 + GTX 3 (15.0 +/- 6.6%). PMID- 12749198 TI - [New method for the observation of gas-production using fiber-stuffed tube for coliform detection and EC-test]. AB - Gas production in lactose-containing medium (e.g., BGLB, LB, or EC medium) is the most important characteristic in the E. coli/Coliform group (C.F.G.) test on food and water. Generally a Durham tube is used as the fermentation tube, and the collected gas is analyzed. However, difficulties can arise, such as insufficient gas volume or muddy precipitates. Since air clings well to fibers in water, the Tsunoda tube (T-tube), a stalk-like tube stuffed with synthetic fibers which can capture the gas more precisely and easily, was designed. With this T-tube, air elimination during the preparation of the medium and detection of gas, even in small quantities, are both simplified. Our results show that the T-tube can replace the Durham tube as a device for detecting gas-production by the E. coli/Coliform group, with improved accuracy and sensitivity. PMID- 12749199 TI - Determination of antioxidant activity of herbs by ESR. AB - Water extracts of 32 herbs that are constituents of curry and curry powder were screened for superoxide anion radical (O2.-) scavenging activity. Among the screened samples, only clove, allspice, and basil were shown to decrease DMPO-O2. adduct yields by more than 50% at 0.25 mg/mL as measured by an ESR spin trapping technique based on the HPX-XOD reaction. To study the mechanism of the O2.- scavenging activity, Km values were obtained from a Lineweaver-Burk plot for XOD in the presence of different concentrations of HPX, and the IC50 values at different DMPO concentrations were compared. Clove and basil directly eliminated O2.- like superoxide dismutase (SOD), whereas allspice reduced the amount of O2.- by inhibition of formation of O2.-. PMID- 12749200 TI - [Analysis of imazalil and its major metabolite in citrus fruits by GC-FTD]. AB - A simplified simultaneous analytical method of imazalil (IZ) and its major metabolite, alpha-(2,4-dichlorophenyl)-1H-imidazole-1-ethanol (IZM), in citrus fruits was developed, and commodities samples were investigated. A homogenate of citrus fruits was extracted with ethyl acetate under basic conditions. The crude extract was partitioned between 0.025 mol/L of sulfuric acid and ethyl acetate. The analytes were extracted from the aqueous fraction under basic conditions with ethyl acetate. The extract solution was purified with an ENVI-Carb cartridge, and then analyzed by GC-FTD and GC/MS. Recoveries of IZ and IZM added to grapefruit at the level of 0.05 microgram/g were 90.0 and 108.7%, and those in the case of lemon were 100.4 and 93.0%, respectively. The detection limits were 0.01 microgram/g in samples. By this method, IZ and IZM were analyzed in 46 citrus fruits on the market and were detected simultaneously in some samples. PMID- 12749201 TI - [Determination of residual avoparcin in chicken muscle by HPLC with ultraviolet and amperometric detection]. AB - An analytical method was developed for determination of residual avoparcin in chicken muscle by measuring alpha- and beta-avoparcin, major components of the pharmaceutical preparation avoparcin, using HPLC with UV and amperometric detectors. The analytical HPLC was run on a Cosmosil 5C18-AR column (4.6 mm x 25 cm) with a gradient formed from A: 2.5% acetic acid, 0.01 mol/L sodium heptane sulfonic acid-acetonitrile (88.5:11.5) (pH 4.0) and B: 2.5% acetic acid acetonitrile (10:90), using UV and amperometric detection (AMD) with glassy carbon electrode (+900 mV). Avoparcin was extracted from chicken muscle by homogenization with methanol-0.2 mol/L sulfuric acid (6:4) followed by centrifugation after pH adjustment to 4 with 1 mol/L sodium hydroxide. The supernatant was evaporated to dryness, and the residue was dissolved in water. The aqueous layer was adjusted to pH 4 by adding 1 mol/L sodium hydroxide. Then it was purified on a Sep-Pak tC18 plus ENV cartridge. The cartridge was washed with water, and retained substances were eluted with 50% methanol. The eluate was evaporated to dryness under reduced pressure. The residue was dissolved in water and determined by HPLC. Recoveries of avoparcin spiked in chicken muscle were 73.1-88.1% at levels of 2-10 micrograms/g. The detection limits were 0.5 microgram/g (UV) and 0.2 microgram/g (AMD). PMID- 12749202 TI - [Determination of methanol in gardenia red and gardenia blue]. AB - The natural food color gardenia red is manufactured through the hydrolysis of the methyl ester of iridoid glucoside. Gardenia blue is also made from iridoid glucoside. Therefore, there is a possibility that the commercial products contain methanol. To determine methanol in gardenia red and gardenia blue, a headspace GC method with standard addition was developed. In the case of gardenia blue with the methyl ester, methanol may be formed during the analytical procedures for methanol. Thus, conditions in which methanol would not be produced in the headspace-GC method were investigated. Vials containing 1 g of the color preparation, 1 mL of water, and a standard solution were sealed. Equilibrium temperature was an important factor among the conditions for analyzing methanol in gardenia blue. Although at room temperature and 50 degrees C, the contents of methanol were equal, the content increased 1.2 times at 80 degrees C. Methanol contents determined at 50 degrees C were 8 and 9 micrograms/g in two gardenia red products and 25-34 micrograms/g in three gardenia blue products, which were below the residual limit of 50 micrograms/g set for many other natural food additives. PMID- 12749203 TI - [Properties of agar as media for bacteria cultivation and for electrophoresis]. PMID- 12749204 TI - [Current worldwide regulation for mycotoxins]. PMID- 12749205 TI - [Investigation of the causative agents related to complaints and accidents using SPME-GC/MS]. PMID- 12749206 TI - Maternity legal case reports. PMID- 12749207 TI - Away from the Westminster hothouse. PMID- 12749208 TI - Middle colic artery-gastroepiploic artery bypass for compromised collateral flow in distal pancreatectomy with celiac artery resection. AB - BACKGROUND/AIMS: Radical distal pancreatectomy with en-bloc resection of the common hepatic, celiac, and left gastric arteries for pancreatic body cancer that involves these arteries does not routinely require arterial reconstruction because the collateral pathways via the pancreatoduodenal arcades from the superior mesenteric artery are recruited immediately. However, accidental injury to the pancreatoduodenal artery compromises collateral blood flow and may lead to fatal complications. This article describes the middle colic artery gastroepiploic artery bypass as an emergent salvage procedure for restoring collateral flow. METHODOLOGY: The inferior pancreatoduodenal artery was accidentally injured in 2 of 9 patients who underwent the radical procedure between 1997 and 2001. Microvascular anastomosis between the left branch of the middle colic artery and the gastroepiploic artery in an end-to-side fashion was employed. RESULTS: The pulsation of the gastroepiploic artery and the color of the stomach recovered immediately after completion of the middle colic artery gastroepiploic artery bypass. No ischemia-related complication developed postoperatively. Postoperative angiography showed the middle colic artery gastroepiploic artery bypass supplying arterial flow to the liver, stomach, duodenum, and pancreas. CONCLUSIONS: The middle colic artery-gastroepiploic artery bypass is an excellent alternative restoring compromised collateral flow via the pancreatoduodenal arcades when microsurgical technique is available. PMID- 12749209 TI - Serum levels of vascular endothelial growth factor, basic fibroblast growth factor and endostatin in human metastatic liver tumors. AB - BACKGROUND/AIMS: Angiogenic factors are related to the malignant potential of tumors. This study aimed to investigate the relationship of serum angiogenic factors to liver metastasis. METHODOLOGY: The serum levels of vascular endothelial growth factor, basic fibroblast growth factor and endostatin were measured using EIA in 25 patients with metastatic liver tumors and were compared with those of 12 cancer patients without metastasis and 15 controls. RESULTS: The serum vascular endothelial growth factor concentration was significantly higher in the liver metastasis group (503 +/- 84 pg/mL) than in the no metastasis group (205 +/- 38 pg/mL) and the control group (201 +/- 26 pg/mL). The three groups had similar serum basic fibroblast growth factor concentrations. There was no significant difference in serum levels of endostatin among the liver metastasis group (18.8 +/- 1.5 ng/mL), the no metastasis group (23.9 +/- 4.9 ng/mL), and the control group (17.1 +/- 1.5 ng/mL). CONCLUSIONS: Angiogenic response is more prominent than anti-angiogenic responses in liver metastasis. These findings support the rationale for anti-angiogenesis therapy such as endostatin therapy in patients with liver metastasis. PMID- 12749210 TI - Extended operation with or without intraoperative (IORT) and external (EBRT) radiotherapy for gallbladder carcinoma. AB - BACKGROUND/AIMS: Gallbladder carcinoma is a rare disease with dismal prognosis. However, lately improved survival has been reported after extended operation including liver resection and lymphadenectomy in addition to cholecystectomy. The aim of this study was to evaluate such a surgical strategy with and without adjuvant intra- and postoperative radiotherapy (IORT/EBRT). METHODOLOGY: 20 patients underwent extended operation and the last 10 of them IORT/EBRT in addition. Tumor staging was done using the TNM system, determination of histological tumor differentiation and immunohistochemical assessment of p53, Ki67, metallothionein, deleted in colorectal cancer and carcinoembryogenic antigen in tumor tissue. RESULTS: There was no hospital mortality. Postoperative complications occurred in 3 patients (15%). Actuarial 5-year survival was 47% in the radiotherapy group and 13% after operation only (NS). The corresponding figures for median survival are 28.8 and 20.2 months, respectively. Five patients are still alive in the radiotherapy group. There was no difference in tumour stages of the two groups irrespective of the way of evaluation. CONCLUSIONS: The results suggest that extended operation for gallbladder carcinoma +/- IORT/EBRT can be done safely. The tendency to longer survival after adjuvant radiotherapy was not statistically significant. PMID- 12749211 TI - Biliary drainage tube evaluation after common bile duct exploration for choledocholithiasis. AB - BACKGROUND/AIMS: We studied the postoperative evaluation of transcystic duct tube drainage (C-tube), T-tube drainage (T-tube), and retrograde transhepatic biliary drainage after common bile duct exploration for patients with choledocholithiasis. METHODOLOGY: We analyzed the preoperative clinical features of patients, intraoperative findings, postoperative status and management, daily output of bile, liver function, postoperative infections, and postoperative complications for patients who underwent common bile duct exploration including 16 C-tube, 17 T-tube, and 8 retrograde transhepatic biliary drainage cases. RESULTS: There were no significant differences in the preoperative clinical features, intraoperative findings, or the daily output of bile from the tube. The removal day of the biliary drainage tube and postoperative hospital stay were shorter in the C-tube group than in the T-tube and retrograde transhepatic biliary drainage groups. Aspartate amino-transferase level and body temperature in the C-tube group on day 7 were lower than those in the T-tube group, and the total bilirubin level in the C-tube group on day 14 was lower than in the T-tube and retrograde transhepatic biliary drainage groups. Moreover, postoperative complications occurred significantly less frequently in the C-tube group (25.0%) than in the T-tube group (76.5%). CONCLUSIONS: C-tube drainage is thought to be most useful after common bile duct exploration for patients with choledocholithiasis. PMID- 12749212 TI - Recurrence of primary biliary cirrhosis after orthotopic liver transplantation. AB - BACKGROUND/AIMS: The incidence of graft failure due to recurrence of primary biliary cirrhosis after liver transplantation and the clinical value of histological and serological parameters indicating a recurrence are still discussed controversially in the literature. METHODOLOGY: In a retrospective study 18 patients who had received orthotopic liver grafts for primary biliary cirrhosis were investigated for recurrence of disease. Histological findings, the appearance of primary biliary cirrhosis associated autoimmune diseases, the course of antimitochondrial antibodies, serological parameters of liver function and HLA status were evaluated. RESULTS: During a median follow-up period of 114 months, 6 of 18 patients developed a recurrence of primary biliary cirrhosis as indicated by liver biopsies, one patient developed clinical graft failure together with histological recurrence. Twelve patients developed autoimmune diseases. Antimitochondrial antibodies were present in all patients within a period of 12 months after transplantation. Serological parameters were elevated in 16 of 18 patients. Histological findings, serological parameters and associated diseases did not correlate with each other and with clinical symptoms of primary biliary cirrhosis recurrence. CONCLUSIONS: Graft failure due to recurrent disease can occur, but, despite a follow-up period of nearly 10 years, it is not possible to define predictive factors on the basis of histology or serology. PMID- 12749213 TI - Reduction of the risk of unretrieved stones during laparoscopic cholecystectomy. AB - Laparoscopic cholecystectomy entails the risk of gallbladder rupture and consequent loss of stones within the abdominal cavity, which is not an uncommon complication. The development of intraperitoneal abscesses due to the spilled gallstones is one of its major complications. When gallbladder was injured during laparoscopic cholecystectomy, gallbladder was dissected on the medial and lateral side, or from the fundus of the gallbladder in the original position to reduce the spillage of stones. After putting the removed gallbladder into the endoscopic bags, hepatorenal fossa and right subphrenic space was thoroughly examined using retractor and oblique view scope. We performed these procedures in 30 consecutive patients with gallbladder ruptured during operation. Dropped stones were noted in 5 patients and were retrieved successfully. Reduction of stone spillage and the retrieval of spilled stones were essential. It is advisable to retrieve as many gallstones as possible after gallbladder perforation during laparoscopic cholecystectomy. PMID- 12749214 TI - Biliary stricture with hepatolithiasis as a late complication of retrograde transhepatic biliary drainage. AB - A 63-year-old man was admitted to a community hospital complaining of fever and epigastric pain. He had undergone cholecystectomy and choledocholithotomy with retrograde transhepatic biliary drainage 7 years previously. Referred to our hospital after demonstration of hepatolithiasis by computed tomography, he underwent further imaging that showed a dilated left lateral anterior segmental bile duct (B3) with hepatolithiasis. After he underwent percutaneous transhepatic biliary drainage via the B3 segmental bile duct, cholangiography performed through the drainage catheter revealed a biliary stricture at the confluence of B3 associated with intrahepatic stones. percutaneous transhepatic cholangioscopy showed a stricture and a cholangioscopic biopsy specimen contained no malignant cells. After performing cholangioscopic lithotomy, an endoprosthesis was inserted and connected to a subcutaneously placed reservoir. Repeat percutaneous transhepatic cholangioscopy 10 months later demonstrated a decreased degree of the stricture, so the endoprosthetic catheter could be removed. Retrospective review of computed tomography images obtained just after the first operation indicated that the retrograde transhepatic biliary drainage catheter had passed close to the B3, and that intrahepatic bile duct dilation was not present. Therefore, we suspect that biliary stricture was caused by an old bile duct injury due to retrograde transhepatic biliary drainage catheter placement. Percutaneous transhepatic cholangioscopy effectively managed this stricture and associated hepatolithiasis. PMID- 12749215 TI - Cholangiocarcinoma arising from preexisting biliary hamartoma of liver--report of a case. AB - We describe the case of a 72-year-old asymptomatic man with a cholangiocarcinoma arising from a biliary hamartoma, also referred to as "von Meyenburg's complex". The patient was clinically diagnosed as having a cystadenocarcinoma, but the tumor had already been present as a uniformly low-density area on computed tomography taken four years previously, as revealed by retrospective examination of the computed tomography films that had been taken annually after surgery for pulmonary emphysema. The low-density area had continued to enlarge year after year, and a high-density area was observed to have emerged inside the low-density area on computed tomography. Histopathological examination demonstrated that the high-density area corresponded to the cholangiocarcinoma and the low-density area to a biliary hamartoma. This is the first case in which it was possible to confirm the presence of cholangiocarcinoma inside a biliary hamartoma that had continued to increase in size. PMID- 12749216 TI - Outcome after resection of central cholangiocarcinoma: preliminary experience with 46 patients in a single center. AB - BACKGROUND/AIMS: Despite the enormous progress in surgery the management of cholangiocarcinoma remains a problem. In this study we present our preliminary experience in the surgical treatment of central cholangiocarcinoma. METHODOLOGY: Between May 1997 and December 2001, 175 patients with central cholangiocarcinoma were admitted to our center. Of the 175 patients, 54 cases subjected to surgery, the remaining 131 patients did not undergo surgery because of advanced disease, advanced liver cirrhosis and poor general condition. Forty-six patients underwent surgical excision, their mean age was 53 +/- 3 years, and male to female ratio was 74-26%. All patients presented with jaundice associated with pain (30%) or biliary stones (32%). Surgical resection of the bile duct with or without part of segment IV were done in fourteen patients and bile duct resection together with major hepatectomy had been done in the remaining 28 patients. RESULTS: Hospital mortality occurred in 10.8%; the main cause of mortality was hepatic cell failure. The most common complications were hepatic cell failure that occurred in 7 cases (15%), biliary leakage in 8 cases (17%), gastrointestinal bleeding in 3 cases (6.5%) and wound infection in 3 cases (6.5%). Late complication in the form of recurrence occurred in 12 cases (29.5%). cholangitis in 8 cases (19.5%), hepatic cell failure in 5 (12%) and gastrointestinal bleeding in 2 (4.8%). At the end of the study, 19 patients (46%) were alive with mean follow-up of 16.6 +/- 9 months. The survival rate at 6, 12, 18, 24 months was 92, 82, 52, 25, 18%, respectively. Recurrence and cholangitis were found significantly higher in the group without hepatectomy than the group after hepatectomy (p < 0.0001, p < 0.0016, respectively). CONCLUSIONS: From our results we can conclude that major hepatectomy with excision of the extrahepatic bile duct system and caudate lobe resection may be recommended for the surgical treatment of central cholangiocarcinoma in selected cases. PMID- 12749217 TI - Variations of intrahepatic and proximal extrahepatic bile ducts. AB - BACKGROUND/AIMS: The modern segmental concept of the liver with a segmental bile duct distribution is of practical importance for biliary specialists and should include information on variations of intrahepatic and extrahepatic bile ducts. The aim of this study was to find biliary variations, especially on a segmental and sectorial level, and to try to arrange them into types. METHODOLOGY: We analyzed 51 corrosion casts of the human liver, which enabled the three dimensional study of extrahepatic, sectorial, segmental and subsegmental bile ducts, their variations in course and confluencing, and the relationship between the structures of the portal pedicle. RESULTS: The types of confluence and their frequency were determined separately for the left, right, right anterior sectorial, right posterior sectorial and common hepatic ducts. "Normal" left confluence was formed in 82% and three variations in 18%. "Normal" right confluence was formed in 75% and four variations in 25%. A complete ("normal") anterior sectorial duct was present in 35% and four variations in 65%. A complete ("normal") posterior sectorial duct was present in 61% and four variations in 39%. CONCLUSIONS: The study showed that the biliary tree variations are quite frequent, and therefore clinically important, and that they could be arranged into several types. PMID- 12749218 TI - Octreotide augments the effects of hepatic arterial occlusion in treating a rat liver tumor model. AB - BACKGROUND/AIMS: To observe if octreotide can augment the effects of hepatic arterial occlusion for transplanted cancer in rat's liver. METHODOLOGY: Walker 256 carcinosarcoma was transplanted into rat's liver to create the liver cancer model. Hepatic arterial ligation was used to block the hepatic arterial blood supply. Rats bearing tumor were divided into three groups: control group, HAL (hepatic arterial ligation) group, and HAL plus octreotide group. Change of tumor volume and tumor growth inhibiting rate after therapy were evaluated. Hoechst 33342 labeling assay was used to analyze the blood perfusion of tumor (the labeled cells' number presenting blood perfusion). Expression of vascular endothelial growth factor and matrix metalloproteinase-1 mRNA was detected by in situ hybridization, and the level of serum vascular endothelial growth factor was assayed by ELISA. RESULTS: Six days after hepatic arterial ligation, the mean tumor volume in control group, HAL group, and HAL plus octreotide group was 0.103 +/- 0.043 cm3, 0.030 +/- 0.018 cm3, and 0.016 +/- 0.005 cm3, respectively. The tumor volume in the two behind groups was smaller than that in the control group (P < 0.01), and the tumor growth-inhibiting rate was 70.8%, and 84.5%, respectively. Compared with the HAL group, the tumor volume in HAL plus octreotide group decreased significantly (P < 0.05). Hoechst 33342 labeled cells' number in control group, HAL group, and HAL plus octreotide group was 369.7 +/- 30.2, 344.1 +/- 26.0, and 323.2 +/- 40.4, respectively. The number in HAL combined with octreotide group decreased significantly compared with that in control group (P < 0.05), which suggested that the blood perfusion of tumor in HAL plus octreotide group decreased significantly. The expression of vascular endothelial growth factor and matrix metalloprotenase-1 mRNA decreased slightly, but not significantly in HAL plus octreotide group compared with that in HAL group (P > 0.05). CONCLUSIONS: The results suggest that octreotide can promote the effects of hepatic arterial occlusion therapy for transplanted cancer in rat's liver. Decreasing the blood perfusion of tumor after hepatic arterial blockage maybe one of its major mechanisms. PMID- 12749219 TI - C-reactive protein and gallium scintigraphy in patients after abdominal surgery. AB - BACKGROUND/AIMS: Early detection of post-surgical infection is important to decrease mortality in patients after operation. Both C-reactive protein test and gallium-67 scan (gallium scan) are sensitive examinations in the detection of infection. In this study, we compared the diagnostic accuracy of the two modalities in the detection of infection after abdominal surgery. METHODOLOGY: Forty-six patients undergoing abdominal surgery were enrolled in this study. All patients received blood examination for C-reactive protein test and were referred to our department for gallium scan because of unknown fever after surgery. RESULTS: Of the 46 patients with abdominal surgery, 22 (47.8%) were diagnosed to have infection including 6 intra-abdominal abscesses, 8 wound infection and 8 with both intra-abdominal abscesses and wound infection. To achieve better diagnostic results, C-reactive protein value of 2.8 mg/dL was chosen as cut-off value. The diagnostic sensitivities for both gallium scan and C-reactive protein test were 100%. The diagnostic specificity of gallium scan was superior to C reactive protein test (83.3% vs. 54.2%). The overall diagnostic accuracy of gallium scan and C-reactive protein test were 92.6% and 76%, respectively. CONCLUSIONS: Both C-reactive protein test and gallium scan have good sensitivity in the detection of infection after abdominal surgery. Gallium scan has better diagnostic specificity than the C-reactive protein test. PMID- 12749220 TI - High incidence of colic anastomotic leakage complicating upper abdominal en bloc evisceration for cancer: a 47-patient series. AB - BACKGROUND/AIMS: The purpose of this study was to analyze the early outcome of en bloc extended resection for upper abdominal locally advanced cancer. METHODOLOGY: A retrospective medical chart review was performed in 47 consecutive patients who underwent an upper abdominal en bloc resection for cancer involving multiple organs or structures at Paoli-Calmettes Institute and Conception Hospital from October 1988 through April 1997. A third of patients underwent a resection of 4 sus mesocolic organs or more. RESULTS: The postoperative morbidity and mortality rate were respectively, 57% and 19%. Despite a high number of theoretically risky procedures including pancreatic resection and pancreatojejunostomy, total gastrectomy and esophagojejunostomy, total hepatectomy and liver transplantation, the higher percentage of complication was found with colic anastomosis. Five of the 30 patients (17%) who underwent a colic anastomosis developed a colic anastomotic leakage, 4 patients were reoperated and 2 patients died. CONCLUSIONS: Because of this unacceptably high rate of complications, we propose to systematically perform a protective stoma when an upper abdominal evisceration includes a colic anastomosis. PMID- 12749221 TI - Resected case of a double cancer, a hepatocellular carcinoma and a cholangiocellular carcinoma, and their spread to the skin. AB - Combined hepatocellular and cholangiocellular carcinomas are rare. Moreover, double cancer cases of hepatocellular carcinoma and cholangiocellular carcinoma are very rare. This report describes a patient with double cancer. A correct clinical diagnosis was made with successful resection, and cutaneous metastases occurred near the exit site of an abdominal drain after the resection of the tumor. The patient, a 66-year-old man with chronic hepatitis C, was admitted to our hospital because he was suspected of having primary liver cancer. Two liver masses in the anteroinferior segment were detected by using angiography, computed tomography during angiography, and computed tomography during arterioportography. These clinical findings indicated that the tumor in the right lobe was hepatocellular carcinoma. A resection of the S5 subsegmentectomy was performed. One mass was diagnosed histologically as hepatocellular carcinoma, and the other mass was diagnosed as cholangiocellular carcinoma. One year after the operation, the patient palpated a hard subcutaneous nodule 4.0 cm in diameter in the right lower abdominal wall. A subcutaneous tumor was excised, and a histological examination revealed moderately differentiated hepatocellular carcinoma. The patient is currently doing well without further recurrence of hepatocellular carcinoma or cholangiocellular carcinoma, 18 months after subsegmentectomy and six months after excision of the subcutaneous tumor. PMID- 12749222 TI - Autofluorescence endoscopy images of pancreas cancer: report of a case. AB - This is the first report of the observation of pancreas cancer with an autofluorescence endoscopic imaging system (excitation: 437 nm). A case of intraductal papillary adenocarcinoma of pancreas was presented. After pancreatectomy, the resected pancreas was used to test the endoscope (16Fr) in the pancreatic duct. The normal pancreatic duct was seen as light blue and the protruding cancerous lesion was observed as a dark red image. In previous studies, cancerous lesions of the gastrointestinal tract, bronchial tree and bile duct also appeared dark red when examined by autofluorescence endoscopy. In the pancreatic duct, the cancer lesion was also detected as dark red color. PMID- 12749223 TI - Multilocular cystic hepatocellular carcinoma (CHCC) mimicking mucinous cystadenocarcinoma. AB - Hepatocellular carcinoma with cystic formation is very rare. Only 5 cases have been reported previously in the literature. We present a 55-year-old man with a symptomatic large cystic hepatic mass. Both abdominal ultrasound and computed tomography revealed a multilocular cystic mass with solid component, occupying most of the left lobe of the liver. An extended left hepatectomy was performed on the basis of a presumptive diagnosis of a cystadenocarcinoma. Pathologic and immunohistochemical evaluation however confirmed a cystic hepatocellular carcinoma without cirrhosis. The incidence and prognosis of this rare entity is unknown. PMID- 12749224 TI - Carcinoid of the papilla of Vater; a case report. AB - A 68-year-old Japanese man, without any symptoms, was found to have a carcinoid tumor of the Ampulla of Vater. A physical examination indicated no anemia or jaundice and no abnormal findings at all in the chest or abdomen. Except for glucose intolerance, the routine laboratory data were normal. An endoscopic biopsy was performed that suggested malignant tumor cells. There were no signs of carcinoid syndrome. A pylorus-preserving pancreatoduodenectomy with extensive lymph node dissection was performed. Histological and immunohistochemical studies resulted in the diagnosis of a carcinoid of the papilla of Vater, without regional lymph node metastases. Although postoperative, an anastomotic leakage of pancreaticogastrostomy was noted; the pancreatic fistula was closed seven weeks later to use the somatostatin analogue. PMID- 12749225 TI - Extended right hepatectomy and total gastrectomy with double Roux-en-Y reconstruction for hilar bile duct and gastric carcinomas. AB - A 70-year-old man with double carcinomas of the hilar bile duct and stomach underwent extended right hepatectomy and total gastrectomy with double Roux-en-Y reconstruction in a one-stage procedure 15 days after right portal embolization. The postoperative course was uneventful, and he was discharged 33 days after surgery. This report describes details of the surgical techniques employed in this patient. PMID- 12749226 TI - Extended abdominoperineal resection with partial prostatectomy for T3 rectal cancer. AB - BACKGROUND/AIMS: To describe and discuss extended abdominoperineal resection with partial prostatectomy using the ultrasonic harmonic scalpel as an alternative to total pelvic exenteration in lower rectal T3 cancer contiguous with the prostate. METHODOLOGY: Extended abdominoperineal resection with partial prostatectomy was performed in 4 patients with lower rectal clinical stage T3 cancer contiguous with the prostate using the ultrasonic harmonic scalpel. The ultrasonic harmonic scalpel permitted en bloc dissection of the rectum and the posterior part of the prostate, removing all tumor and disruption of the normal urinary stream. RESULTS: Blood loss ranged from 600 to 2500 mL. The final pathologic stage was T2 in 1, T3 in 2 and T4 in 1 patient, and the surgical margins were clear in all patients. Urethral injury occurred in 1 patient, and 2 patients had postoperative urinary dysfunction. No patients required a urinary stoma or suffered recurrence. CONCLUSIONS: Extended abdominoperineal resection with partial prostatectomy using the ultrasonic harmonic scalpel is an attractive alternative to total pelvic exenteration for patients with lower rectal T3 cancer contiguous with the prostate. PMID- 12749227 TI - Effects of idiotypic human anti-mouse antibody against in vitro binding and antitumor activity of a monoclonal antibody-drug conjugate. AB - BACKGROUND/AIMS: Because the HAMA (human anti-mouse antibody) response following administration of murine monoclonal antibodies represents mainly isotypic HAMA production, idiotypic HAMA responses have not been thoroughly analyzed. METHODOLOGY: In the present study we examined the effect of idiotypic HAMA that arose in patients who had repeatedly received murine monoclonal antibody conjugated to an anticancer drug against in vitro binding and antitumor activity of the conjugate. HAMA that had developed after administration of tumor-specific murine monoclonal antibody A7 conjugated with neocarzinostatin were measured in patient serum. The inhibitory effect of HAMA on tumor binding and antitumor activity of the A7-neocarzinostatin conjugate was examined in cultures of human colonic carcinoma cells. RESULTS: The serum concentration of idiotypic HAMA in patients administered A7-neocarzinostatin was significantly higher than that in a control group. Binding activity and antitumor activity of A7-neocarzinostatin against target cells was reduced in the presence of sera containing HAMA. CONCLUSIONS: Repeated use of A7-neocarzinostatin can be expected to show less antitumor effect than the first use of the conjugate. PMID- 12749228 TI - Surgical strategy to save patients with colon perforation with chronic renal failure on long-term hemodialysis. AB - BACKGROUND/AIMS: The aim of the current study is to clarify a strategy to save patients with diffuse peritonitis due to colonic perforation, who have been undergoing long-term hemodialysis with chronic renal failure. METHODOLOGY: Five patients with colon perforation with chronic renal failure on long-term hemodialysis were studied and their clinical courses and outcomes after surgical treatment were investigated. RESULTS: The proportion of salvage for patients in our cases was 60%. The chief reasons of failure in salvage in two patients were mainly a delay of surgical treatment and the compromised condition with recurrence of the neoplasm. In the current study, level of serum concentration of C-reactive protein of the patients on admission was found not to be reliable to decide a surgical indication, and delay of the surgical treatment was fatal. CONCLUSIONS: The physical examination is most important and reliable to decide the surgical indication for patients with colon perforation, and even with suspicion of peritonitis requiring a laparotomy, an immediate surgical treatment should be courageously performed for patients with chronic renal failure on long term hemodialysis. PMID- 12749229 TI - Budding as a useful determinant of the optimal treatment for T1 rectal carcinomas. AB - BACKGROUND/AIMS: If a locally excised specimen has poorly differentiated or undifferentiated histology, positive vascular invasion, or massive invasion to the cut end or a positive margin, additional surgery is recommended for the treatment of T1 rectal carcinoma. However, positive predictive values of these histological criteria are low. This study was undertaken to clarify more reliable risk factor(s) for lymph node metastasis in T1 rectal carcinomas. METHODOLOGY: In 58 patients with T1 rectal carcinoma undergoing local excision or radical surgery, the associations between lymph node metastasis or intrapelvic recurrence and clinicopathological features were studied using multiple regression analysis with special reference to tumor budding at the invasive front. RESULTS: Lymph node metastasis was observed in 1 of 9 patients undergoing additional bowel resection after local excision, and in 2 of 24 patients undergoing radical surgery alone. Intrapelvic extrarectal recurrence was observed in 3 of 25 patients undergoing local excision alone. Logistic regression analysis revealed that budding at the invasive front alone was significantly associated with lymph node metastasis or intrapelvic recurrence (p = 0.0484). CONCLUSIONS: Additional bowel resection with lymph node dissection should be recommended for locally excised T1 rectal carcinoma with budding at the invasive front. PMID- 12749230 TI - Mean corpuscular volume (MCV) and the risk of colorectal adenoma in menopausal women. AB - BACKGROUND/AIMS: It has been reported that alcohol intake and folate deficiency are associated with an increased risk of colorectal adenomas and carcinomas. Mean corpuscular volume is increased under these conditions. We have reported that the mean corpuscular volume was higher in patients with adenoma than without adenoma in middle-aged men. The aim of this study was to assess the association between mean corpuscular volume and risk of colorectal adenoma in menopausal women. METHODOLOGY: The subjects were 415 menopausal women who underwent both barium enema examination and total colonoscopy, and their blood samples were analyzed. The subjects were divided into two groups with or without adenoma, and were divided into four groups according to the mean corpuscular volume value. Various variables were compared among the groups, and the odds ratios of adenoma were calculated. RESULTS: The mean corpuscular volume was higher in patients with adenoma than without adenoma (P = 0.002). As for the mean corpuscular volume value, the odds ratio (95% CI) of patients with adenoma was 1.00 (referent); (mean corpuscular volume (fl) < 90), 1.50 (0.93-2.07); (> or = 90 but < 92.5), 1.52 (0.97-2.07); (> or = 92.5 but < 95) and 2.87 (2.25-3.45); (> or = 95). CONCLUSIONS: Mean corpuscular volume > or = 95 may be used as an index of the risk for colorectal adenomas in menopausal women. PMID- 12749231 TI - Oncologic outcome of laparoscopic surgery for T1 and T2 colorectal carcinoma. AB - BACKGROUND/AIMS: Laparoscopic surgery for colorectal carcinoma remains controversial because of the technical difficulties and uncertainty regarding the long-term oncologic outcome after laparoscopic surgery. The objective of this study was to evaluate the feasibility for the laparoscopic surgery in the treatment of pT1 and pT2 colorectal carcinoma. METHODOLOGY: A review was performed of a prospective registry of 226 patients who underwent curative laparoscopic resection for pT1 and pT2 colorectal carcinoma between December, 1992 and December, 2001. Patient demographics and outcomes were recorded prospectively. RESULTS: The median follow-up was 43 months. Three patients (2.0%) in the pT1 group and 3 patients (3.9%) in the pT2 group developed recurrence of carcinoma. The expected five-year survival and disease-free survival rates in the pT1 group were 98.9% and 97.6%, respectively, whereas they were 93.6% and 93.4% in the pT2 group. No patient had port-site or peritoneal recurrence during the follow-up period. CONCLUSIONS: The findings of current study demonstrate that oncologic outcome of laparoscopic surgery for patients with pT1 and pT2 colorectal carcinoma appear to be comparable with conventional surgery. Laparoscopic surgery is oncologically appropriate at least for patients with pT1 and pT2 colorectal carcinoma. PMID- 12749232 TI - A case of duodenal duplication in an adult. AB - We report a rare case of duodenal duplication, manifested in an adult with aspecific symptoms, whose diagnosis was possible with ultrasound at the beginning and then with gastrointestinal barium study, endoscopic ultrasonography and contrast enhanced CT scan; it was confirmed surgically. PMID- 12749233 TI - 3p21, 5q21, 9p21 and 17p13.1 allelic deletions are potential markers of individuals with a high risk of developing adenocarcinoma in Barrett's epithelium without dysplasia. AB - BACKGROUND/AIMS: A common genetic abnormality detected in Barrett's adenocarcinoma is LOH (loss of heterozygosity) at the sites of known or putative tumor suppressor genes. Thus, some deletions have also been determined in peritumoral Barrett's epithelium. These findings suggest that a tissue field of somatic genetic alterations precede the histopathological phenotypic changes of carcinoma. We investigated 32 cases of Barrett's esophagus with no evidence of dysplasia for LOH at 5q21 (APC), 3p21, 9p21 (p16) and 17p13.1 (p53) chromosomal regions. METHODOLOGY: Two groups were randomly selected and compared: 16 cases of Barrett's epithelium adjacent to adenocarcinoma and 16 cases of Barrett's epithelium with no evidence of malignant transformation in a 5-10 years follow-up period. In three adenocarcinomas cases several previous endoscopic biopsies of Barrett's esophagus were available. RESULTS: We determined frequent allelic losses in adenocarcinomas at p53 (54%), p16 (50%), 3p21 (40%) and 5q21 (33%). Identical LOH was present in most cases in the Barrett's epithelium adjacent to adenocarcinoma. LOH at these loci was unusual in Barrett's epithelium with no evidence of malignant transformation. However, in cases where sequential endoscopic biopsies were performed in advance to the adenocarcinoma diagnosis LOH was already present in the Barrett's epithelium. CONCLUSIONS: We suggest that LOH at these loci may be present before the onset of the malignant growth and LOH studies may supplement the histopathological evaluation of Barrett's epithelium. LOH at 3p21, 5q21, 9p21 and 17p13 chromosomal regions in cells of Barrett's epithelium without dysplasia may have a role as a potential marker for individuals with a high risk of developing adenocarcinoma. PMID- 12749235 TI - Anti-neutrophil cytoplasmic antibodies in Brazilian patients with inflammatory bowel disease. AB - BACKGROUND/AIMS: Anti-neutrophil cytoplasmic antibody has been observed in the sera of patients with inflammatory bowel disease, but its prevalence depends on the population being studied and the method employed for its detection. METHODOLOGY: We evaluated the prevalence of antineutrophil cytoplasmic antibody by immunofluorescence assay in a series of Brazilian patients with inflammatory bowel disease: 40 patients with ulcerative colitis and 36 with Crohn's disease. We also correlated the presence of this antibody with duration of symptoms, site of the disease and inflammatory activity. Thirty healthy individuals comprised the control group. RESULTS: Anti-neutrophil cytoplasmic antibody was detected in 27.5% of the patients with ulcerative colitis, and in 14.3% of those with Crohn's colitis. Perinuclear staining pattern was the most common, but atypical and nuclear patterns were also observed. There was no correlation between the presence of this antibody and any of the studied clinical variables. No patient of the control group presented positive test. CONCLUSIONS: A positive anti neutrophil cytoplasmic antibody test would not be helpful in the differential diagnosis between ulcerative colitis and Crohn's colitis, since it has a moderate specificity of 86% for ulcerative colitis patients. PMID- 12749236 TI - The usefulness of technetium-99m hexamethylpropyleneamineoxide labeled white blood cell abdomen scan to differentiate periappendiceal abscess from acute appendicitis--a preliminary report. AB - BACKGROUND/AIMS: The purpose of this study is to assess the usefulness of Tc HMPAO (technetium-99m hexamethylpropyleneamineoxide) labeled white blood cells abdomen scan to differentiate acute appendicitis from periappendiceal abscess. METHODOLOGY: Forty-five patients with atypical symptoms and signs of appendicitis were included in this preliminary study. At 4 hours after an intravenous injection of Tc-HMPAO labeled white blood cells, static images over the anterior abdomen were obtained using a gamma camera. Two regions of interest were selected, one was over the lumbar spine and the other was over the appendiceal area. The mean count ratio of the appendiceal area over the mean pixel count of the lumbar spine was calculated. Sixteen patients were diagnosed with acute appendicitis and 19 patients with periappendiceal abscess by final operative findings. Meanwhile, the other 10 patients with acute abdomen but with no appendicitis served as controls. RESULTS: The mean count ratio of controls, acute appendicitis and periappendiceal abscess was 0.65 +/- 0.05, 0.82 +/- 0.07, and 1.25 +/- 0.11, respectively. If the mean count ratio > or = 1.00 was defined as the cutoff value to diagnose periappendiceal abscess, the specificity and the positive predictive value was 100%. CONCLUSIONS: We concluded that Tc-HMPAO white blood cells abdomen scan should be a potential tool to differentiate periappendiceal abscess from acute appendicitis. PMID- 12749234 TI - Gastric tube cancer after esophagectomy for esophageal squamous cell cancer and its relevance to Helicobacter pylori. AB - BACKGROUND/AIMS: Improvement of surgical skills and postoperative management has allowed longer postoperative survival for patients with esophageal cancer, among those some develop gastric tube cancer. We analyzed the characteristics of such patients we encountered as well as of reported cases of Japan. Furthermore, we investigated if Helicobacter pylori plays a role in carcinogenesis of the gastric tube in our cases. METHODOLOGY: We analyzed the clinicopathological features of our 8 patients with gastric tube cancer from 1991 to 2000 as well as the status of H. pylori on the gastric tube biopsy. Moreover the features of gastric tube cancer from domestic reported cases up to the year 2000 were also summarized. RESULTS: According to the review of our cases, the frequent tumor location was the distal portion of the gastric tube. Seventy-eight percent were detected in early stage during postoperative follow-up, 71% of those were treated endoscopically. No cases showed H. pylori positivity. From the previous domestic reports, early cancer is increasing as the screening becomes popular. Type 0-IIa and 0-IIc were the most popular images for early cancer, while type 2 and 3 were for advanced cancer. CONCLUSIONS: The carcinogenesis of the gastric tube seemed not to be related to H. pylori. Subdermal route of reconstruction at esophagectomy seemed superior regarding early recognition of gastric tube cancer and easiness of its treatment. PMID- 12749237 TI - Elevated serum interleukin 6 levels in patients with acute intestinal ischemia. AB - BACKGROUND/AIMS: Early diagnosis of patients with acute intestinal ischemia may be possible by measuring serum cytokine levels. METHODOLOGY: Forty-six patients presenting in emergency with an acute abdomen where intestinal ischemia was a possible diagnosis were evaluated. A single blood sample was collected in emergency prior to any intervention and the patients were then followed prospectively. Serum tumor necrosis factor-alpha and interleukin-6 levels were determined at a later date. Serum levels in patients with proven acute intestinal ischemia were compared to patients with other diagnoses. RESULTS: Serum tumor necrosis factor-alpha levels were moderately increased in patients with acute intestinal ischemia compared to controls 96.9 +/- 98.9 pg/mL vs. 60.8 +/- 63.7 pg/mL, P = 0.16. Serum interleukin-6 levels were significantly increased in patients with acute intestinal ischemia, 15.778 +/- 21.349 pg/mL vs. 2.844 +/- 5.625 pg/mL, P = 0.01. CONCLUSIONS: Serum interleukin-6 levels may prove useful in diagnosing patients with acute intestinal ischemia. PMID- 12749238 TI - Validation of endoscopic 13C-urea breath test with nondispersive infrared spectrometric analysis in the management of Helicobacter pylori infection. AB - BACKGROUND/AIMS: 13C-EUBT (endoscopic 13C-urea breath test), that combines the conventional urea breath test with endoscopy, provides excellent accuracy for detection of H. pylori. Recently, a simpler, less expensive and isotope-selected nondispersive infrared spectrometer has been developed for the urea breath test. We validated the diagnostic performance of 13C-EUBT using nondispersive infrared spectrometer in the management of H. pylori infection. METHODOLOGY: EUBT was performed in 232 patients by first collecting a baseline breath sample followed by upper gastrointestinal endoscopy. A 20-mL aliquot of 13C-urea solution was sprayed over the entire stomach under endoscopic guidance. Breath samples taken 0 and 20 minutes after spraying were analyzed using nondispersive infrared spectrometer. H. pylori infection was assessed by rapid urease test and histology. The cutoff level was determined by a receiver-operating characteristic curve analysis. Forty-four samples were also analyzed by the conventional isotope ratio mass spectrometer to compare results from both analyzers. We also applied the nondispersive infrared spectrometer-based EUBT to evaluate the efficacy of eradication therapy. RESULTS: Employing 2.4 per mL as the best cutoff value, the EUBT yielded an excellent diagnostic accuracy, with a sensitivity of 99% and specificity of 99%. The sensitivity and specificity of the test for assessing eradication therapy were 94% (16/17) and 100% (57/57), respectively. There was a high linear correlation between nondispersive infrared spectrometer and isotope ratio mass spectrometer (r = 0.998, p < 0.0001). CONCLUSIONS: 13C-EUBT using nondispersive infrared spectrometer is a highly accurate and rapid method for the assessment of H. pylori eradication as well as for detecting H. pylori infection. We believe that nondispersive infrared spectrometer gives more rapid and less expensive method for the management of H. pylori infection with the EUBT. PMID- 12749240 TI - The Zollinger-Ellison syndrome with acute bleeding pancreatitis. AB - A 44-year-old Japanese man was diagnosed to have Zollinger-Ellison syndrome. His main complaint was anemia due to gastrointestinal bleeding. After performing gastrointestinal endoscopy, duodenal ulcers were found located in the posterior wall of the duodenal bulbus. Three months before presentation, he had undergone surgery at our hospital due to acute bleeding pancreatitis. A case of Zollinger Ellison syndrome with acute bleeding pancreatitis is rare, and there have so far been few reports of such cases in the English medical literature. PMID- 12749239 TI - Comparison between technetium-99m hexamethylpropyleneamineoxide labeled white blood cell abdomen scan and abdominal sonography to detect appendicitis in children with an atypical clinical presentation. AB - BACKGROUND/AIMS: Diagnosing acute appendicitis in children with equivocal signs and symptoms is usually difficult. The usual approach to the patient is hospital observation and frequent reexamination. However, many surgeons are reluctant to delay surgery because of the risk of perforation and a negative laparotomy. The aim of this study is to assess and compare the value of the Tc-99m HMPAO (technetium-99m hexamethylpropyleneamineoxide) labeled white blood cell abdomen scan and abdominal sonography in the diagnosis of acute appendicitis in children with an atypical clinical presentation. METHODOLOGY: Forty children with acute abdomen and possible acute appendicitis but atypical findings were included in this study. After an intravenous injection of Tc-99m HMPAO white blood cells, serial anterior abdomen scans at 30 min, 60 min, 120 min and 240 min were obtained using a gamma camera. Meanwhile, at the point of maximal tenderness, abdominal sonography was performed with a graded compression technique for both longitudinal and transverse images. RESULTS: Thirty-three children received operation for surgical and pathological diagnoses. The other 17 children did not receive operation but follow-up of at least a one-month period. The overall sensitivity, specificity, and accuracy of Tc-99m HMPAO white blood cell scan to diagnose acute appendicitis in children with atypical findings is 96.7%, 80.0%, and 90.0%, respectively. The overall sensitivity, specificity, and accuracy for abdominal sonography is 86.6%, 90.0%, and 88.0%, respectively. CONCLUSIONS: The Tc-99m HMPAO white blood cell abdomen scan provides a more sensitive and accurate method for the diagnosis of appendicitis in children with atypical clinical presentation when compared with abdominal sonography. PMID- 12749241 TI - Mucinous cystadenocarcinoma of the appendix. The controversy persists: a review. AB - Mucinous cystic neoplasms of the appendix continue to engender considerable debate in their diagnosis, management, and role in pseudomyxoma peritonei. This review addresses ongoing controversies with these unique neoplasms. Case reports and institutional series of mucinous cystadenocarcinoma were retrieved from the literature using Medline Search (1985-1996) using the following key words: appendix; mucinous cystadenocarcinoma; pseudomyxoma peritonei. 284 patients with mucinous cystadenocarcinoma were identified. Abdominal pain was the most common presenting symptom and objective preoperative diagnosis was difficult. There were 22 patients with concomitant appendiceal and ovarian mucinous neoplasms. Sixty seven patients presented with mucinous cystadenocarcinoma and pseudomyxoma peritonei. However, a unified definition of pseudomyxoma peritonei was lacking. The recurrence rate was higher when an appendiceal neoplasm was associated with pseudomyxoma peritonei. In the reported literature, about an equal number of patients were subjected to appendectomy alone or right hemicolectomy. However, there was a survival advantage for right hemicolectomy versus appendectomy alone. Management of disseminated pseudomyxoma peritonei remains controversial. Mucinous cystadenocarcinoma appears to be a spectrum of low-grade malignant tumors. For correct diagnosis, a high index of suspicion must be maintained. The appropriate treatment for a patient with mucinous cystadenocarcinoma should be a right hemicolectomy. Aggressive debulking may improve survival when associated with pseudomyxoma peritonei. A better definition of pseudomyxoma peritonei is needed. In evaluation of different treatment modalities and prognosis, patients should be grouped according to underlying primary pathology. PMID- 12749242 TI - Effects of portal vein embolization before major hepatectomy. AB - BACKGROUND/AIMS: Major hepatectomy can now be successfully performed after portal vein embolization, but the effects of portal vein embolization have not been clearly delineated. Our objective is to examine whether portal vein embolization really contributes to the success of major hepatectomy. METHODOLOGY: Thirty-eight patients underwent portal vein embolization and hepatectomy of two subsegments or more. They all belonged to a high-risk group according to a prognostic score. We selected 9 of 38 patients with liver metastases (PE-meta group) and 32 patients who had undergone hepatectomy without portal vein embolization (non-PE-meta group) during the study period to compare the serum levels of total bilirubin after hepatectomy. Fifteen of 38 patients had the levels of polymorphonuclear leukocyte elastase and thrombin-antithrombin complex examined after hepatectomy (PE group) and so did 20 patients without portal vein embolization (non-PE group). RESULTS: The maximum levels of total bilirubin in non-PE-meta group correlated with the percentage of hepatic parenchyma to be resected. In the patients receiving portal vein embolization, the pre-PE and post-PE levels were both below the regression. Similar shifts were seen in the graphs of polymorphonuclear leukocyte elastase and thrombin-antithrombin complex. CONCLUSIONS: The effects of preoperative portal vein embolization on safety in major hepatectomy were proved by its suppression of rise in total bilirubin, polymorphonuclear leukocyte elastase and thrombin-antithrombin complex after hepatectomy. PMID- 12749243 TI - Fas-mediated apoptosis in acute alcoholic hepatitis. AB - BACKGROUND/AIMS: The Fas-mediated apoptosis pathway has been implicated in liver diseases. The aim of the study was to investigate the role of this system in alcoholic hepatitis. METHODOLOGY: The expression of Fas, Fas ligand, and intracellular signaling molecules for apoptosis were determined by immunoblot analysis in fresh frozen liver samples from 19 patients with alcoholic liver disease. RESULTS: Fas and Fas ligand expression was significantly increased in the liver of patients with alcoholic hepatitis (n = 11) as compared with alcoholic liver disease patients without hepatitis (n = 8). Similarly, there were significant differences in the expression of FADD, ICE, and CPP32 in the liver between the two groups. There were significant positive correlations between the Fas ligand and the FADD, ICE, or CPP32 levels in the liver. The expression of Fas, Fas ligand, FADD, ICE, or CPP32 correlated with serum markers of hepatic injury. Plasma soluble Fas levels in patients with alcoholic hepatitis (median: 15.1 U/mL, range: 9.7-19.2 U/mL) were significantly higher than in normal controls (n = 9) (median: 2.8 U/mL, range: 1.9-3.7 U/mL; p < 0.001). There was a significant positive correlation between plasma soluble Fas levels and the hepatic expression of FADD in these patients. CONCLUSIONS: These results indicate that Fas-mediated apoptosis may play an important role in alcoholic hepatitis. PMID- 12749244 TI - High-dose interferon-alpha 2b plus ribavirin combination therapy for GB virus C/hepatitis G virus infection--a study in patients with chronic hepatitis C. AB - BACKGROUND/AIMS: The efficacy of interferon-alpha and ribavirin combination therapy for GB virus C/hepatitis G virus infection is not well understood. We previously conducted a double-blind, placebo-controlled trial using high-dose interferon-alpha 2b with or without ribavirin for patients with interferon-alpha relapsed chronic hepatitis C. METHODOLOGY: Fifty-two patients were randomly assigned and completed the 24-week treatment of interferon-alpha 2b (6 million units three times per week) plus ribavirin (1000 to 1200 mg/daily), or plus a matched placebo. Patients were then followed for an additional 24 weeks. RESULTS: Of the 52 patients, 5 patients (9.6%) had hepatitis G virus viremia before or during enrollment. Two patients received interferon-alpha 2b alone and three patients received interferon-alpha 2b and ribavirin combination therapy. At the end of treatment, all of the 5 patients had undetectable hepatitis G virus RNA in sera. Early loss of hepatitis G virus RNA at week 4 of treatment was observed in the 2 patients on combination therapy. Hepatitis G virus RNA reappeared at the end of follow-up in these 5 patients. CONCLUSIONS: In conclusion, interferon alpha 2b and ribavirin combination therapy could induce earlier loss of hepatitis G virus RNA than interferon-alpha 2b alone. Either interferon-alpha 2b alone or interferon-alpha 2b and ribavirin combination therapy achieved transient but not sustained virological response to hepatitis G virus viremia. PMID- 12749245 TI - Percutaneous transhepatic metallic stent placement for malignant portal vein stenosis. AB - BACKGROUND/AIMS: We investigated the safety and efficacy of percutaneous transhepatic angioplasty with self-expanding metallic stent implantation to treat malignant portal vein stenosis. METHODOLOGY: Since 1995, we have performed metallic stent implantation in 7 cases of malignant portal vein stenosis (cholangioma, 3 cases; and cancer of the hepatic hilum, pancreatic cancer, metastatic liver tumor, and hepatocellular carcinoma, 1 case each) and in 1 case of obstruction by local recurrence of a hilar tumor. In the 7 cases of portal vein stenosis, a Wallstent was implanted percutaneously transhepatically. In the case of portal vein obstruction, a Gianturco-Rosch Z stent was implanted via the ileocolic vein during laparotomy. RESULTS: There were no complications related to stent placement, and the portal vein pressure decreased after stent implantation in all cases. The patency of the stent was documented by ultrasonography in all 7 cases of portal vein stenosis and was confirmed at autopsy in two cases. No patient treated for portal vein stenosis developed symptoms of portal hypertension. CONCLUSIONS: Percutaneous transhepatic angioplasty with stent insertion is a safe and useful treatment for malignant portal vein stenosis and preserves quality of life by preventing symptoms of portal hypertension. PMID- 12749246 TI - Insulin resistance in patients with steatohepatitis. AB - BACKGROUND/AIMS: Steatohepatitis is a disease characterized with liver damage due to fat accumulation in more than 5% of the liver weight associated with alcohol intake (consumption) and other (various) causes. However, alcoholic and non alcoholic steatohepatitis do not differ histologically. The aim of study was to investigate whether or not insulin resistance exists in patients with steatohepatitis. Further investigations are needed to show what the mechanisms leading to insulin resistance are. METHODOLOGY: We used a hyperinsulinemic euglycemic clamp to tissue sensitivity to insulin in 11 patients (10 males and one female) and 10 healthy age- and sex-matched individuals. A diagnosis of steatohepatitis was made by clinical, laboratory and histological findings. RESULTS: The amount of glucose infused which is required to keep euglycemia (M value) was markedly low in the study group than in the controls (2.24 +/- 0.33 mg/kg/min and 7.45 +/- 0.82 mg/kg/min, respectively p < 0.0001) suggesting insulin resistance (M value < 4 mg/kg/min). CONCLUSIONS: Our findings demonstrate that insulin resistance does exist in patients with steatohepatitis and the pathophysiological mechanisms causing resistance is not clear but insulin action is an issue of further investigation. PMID- 12749248 TI - High initial dose combination regimen with interferon-alpha and ribavirin in chronic hepatitis C. AB - BACKGROUND/AIMS: Combination of interferon-alpha and ribavirin becomes the antiviral therapy of choice for chronic hepatitis C. The aim of this trial was to assess the efficacy of two combination treatment strategies: standard regimen Interferon-alpha 3 MU three times per week (group A) and initial high dose Interferon 6 MU daily for 2 weeks followed by intermittent administration 3 MU three times per week (group B), plus Ribavirin in naive and relapsed patients with chronic hepatitis C. METHODOLOGY: Twenty-four patients (group A, 6; group B, 18) received medication for median 6 months (range, 5-12) and followed for 6 months. Primary end-point of therapy was a sustained virologic response. Statistical analysis was performed with a t test or non-parametric Mann-Whitney and chi-square test. RESULTS: At the end of the follow-up the overall sustained response rate was 42%: 6 of 18 (33.3%) patients with induction and 4 of 6 (66.6%) with standard regimen. Patients with sustained virologic response received higher interferon dose (423.7 +/- 102.2 MU) at extended duration of therapy (9.8 +/- 2.9 months) versus patients with non-sustained response (p < 0.05). The difference in sustained response rate in both treatment regimens was nonsignificant. CONCLUSIONS: High initial dose therapy does not enhance virologic response, compared to standard combination regimen. PMID- 12749247 TI - Cimetidine reduces impairment of cellular immunity after transcatheter arterial embolization in patients with hepatocellular carcinoma. AB - BACKGROUND/AIMS: Natural killer cells have an intrinsic ability to recognize and attack some tumor cells and infected cells. Natural killer cell cytotoxicity is depressed in patients with hepatocellular carcinoma. In particular, cell-mediated immunity is impaired after transcatheter arterial embolization, which is an effective therapy for hepatocellular carcinoma. There have been reports that cimetidine can activate cellular immunity and increase the survival time in patients with some cancers. Therefore, we investigated whether cimetidine could improve cellular immunity after transcatheter arterial embolization, especially in relation to natural killer activity. METHODOLOGY: Thirty-four patients with hepatocellular carcinoma and cirrhosis who underwent transcatheter arterial embolization at our hospital were studied prospectively after giving informed consent. The patients were divided into three groups. In group A, 14 patients were administered 800 mg of cimetidine a day. In group-B, 13 patients were administered 40 mg of famotidine or 300 mg of nizatidine a day. In group-C 7 patients did not receive histamine 2 receptor antagonists. Natural killer cell activity in peripheral blood was measured before transcatheter arterial embolization and on days 1 and 7. RESULTS: The % natural killer cell activity on day 1 was 74 in group A, 52 in group B, and 52 in group. The % activity on day 7 was 98 in group A, 71 in group B, and 82 in group C. Cimetidine group showed the significant higher % natural killer cell activity on day 1 (p = 0.032). CONCLUSIONS: Our study raises the possibility that cimetidine has the effect to preserve cell-mediated immune response during transcatheter arterial embolization. PMID- 12749249 TI - Long-term effect of hepatocyte transplantation on fulminant hepatic failure in rats. AB - BACKGROUND/AIMS: Effect of hepatocyte transplantation on long-term survival after fulminant hepatic failure was studied in rats. METHODOLOGY: Sprague-Dawley rats were divided into: Group I (n = 65), intrasplenic hepatocyte transplantation followed by fulminant hepatic failure; Group II (n = 31), intrasplenic saline injection followed by fulminant hepatic failure; Group III (n = 24), 70% hepatectomy. For survival, 35 animals of Group I and 19 of Group II were observed. Six animals of each group were euthanized on postoperative days 1, 7, 14 and 28 to study biochemistry, liver growth rate, labeling index of proliferating cell nuclear antigen, hepatocyte growth factor and transforming growth factor. RESULTS: Postoperatively, Group I had a better survival than Group II. Group I also showed a better biochemical profile on day 1 as compared with Group II, and on day 28, Group I had a normal profile. On day 28, the remnant liver in Group I reached 97% of the original liver weight. Group I had a better proliferating cell nuclear antigen labeling index than Group II on day 1 and exceeded Group III on day 14. On day 1, Group I had lower levels of hepatocyte growth factor and transforming growth factor than Group II while hepatocyte growth factor on days 7, 14 and 28 showed no difference between Group I and Group III. CONCLUSIONS: Hepatocyte transplantation has achieved a long-term survival and improved the liver regeneration in rats with fulminant hepatic failure. PMID- 12749250 TI - Repeated liver resection for recurrent metastases from colorectal cancer. AB - BACKGROUND/AIMS: Repeat hepatectomy is the most effective treatment for recurrent colorectal liver metastases. We aim to assess how repeated liver resections increase survival, without unacceptable surgical risk. METHODOLOGY: Between December 1992 and December 1998, among 19 patients, 5 underwent secondary resection of recurrent metastatic disease. Following the primary liver surgery, three patients had systemic chemotherapy with 5-fluorouracil and two locoregional chemotherapy via Port-a-cath in the gastroduodenal artery. We evaluated survival and we compared time of surgery, duration of Pringle maneuver, blood losses and postoperative stay in the hospital between first and second liver surgery. RESULTS: Perioperative mortality at second liver resection was nil; morbidity minor; mean duration of surgery 320 vs. 260 min; Pringle maneuver 35 vs. 25 min; blood losses 1300 vs. 650 mL; postoperative stay 12.6 vs. 11.5 days. Mean total survival from time of colon resection was 50 months. As an interesting secondary finding, we observed prolonged inhibition of liver regeneration following treatment with Methotrexate. CONCLUSIONS: Repeated hepatic resection is a safe procedure for selected patients. Surgical risk is slightly increased, but the risk/benefit ratio is definitely in favor of as many repeated resections as needed, whenever there is a chance of curative surgery. PMID- 12749251 TI - Clinical significance of cell differentiation in hepatocellular carcinoma. AB - BACKGROUND/AIMS: Hepatocellular carcinoma is commonly found in Asian countries and prognosis still remains unsatisfactory due to recurrence after surgical tumor resection. METHODOLOGY: We try to demonstrate the recurrence and survival time in 99 surgical patients grading by tumor cellular differentiation from surgical specimen. RESULTS: The rates of well, moderate, and poor differentiation were encountered in 21 cases (21.2%), 61 cases (61.6%) and 17 cases (17.7%), respectively. Small tumor (< 3 cm) was found in only one (5.9%) in the poor differentiation group and 38.1% and 37.7% in the well and moderate differentiation groups. Capsular invasion was found in 13 (61.9%), 39 (63.9%) and 7 (41.1%) in the well, moderate and poor differentiation group, respectively. We found 41.9% (18/43) and 22.4% (13/58) to be tumor free in capsule invasion (-) and (+) after a period of 18.1 and 29.9 months, respectively. However, the recurrent time was 10.6 and 11.3 months, respectively with no significant difference (p > 0.05). Vascular invasion was more frequent in the poor differentiation group (76.5%) than the well (23.8%) and moderate (60.7%) differentiation groups (P < 0.05). We found 23.5% (4/17) and 35% (21/60) to be tumor free but the recurrence time was 6.5 and 14.1 months for the vascular invasion (-) and (+), respectively. The residual median survival times were 6.5 and 14 months after recurrence, respectively. The tumor recurrence rates were 52.7% (11/21), 52.4% (32/61), and 35.5% (6/17) and recurrence times were 11.7, 11.9, and 4.5 months for the well, moderate and poor differentiation group respectively totally. The recurrence time of young age group (< 39 years old) was shorter than the others and there was no patient of well differentiation less than 40 years old. The recurrence time was shorter in the poor differentiation group but there was no significant difference according to age group. The median survival times were 22.2, 22.9, and 9.5 months for each group, respectively. CONCLUSIONS: Differentiation of hepatocellular carcinoma cell had a clinical significance and was found to be positively correlated with the invasive proclivity. The median survival time was longer in both the well and moderate differentiation group, but not in the poor differentiation group. The clinical data revealed that the extended operations performed upon the patients with poor differentiation effected the recurrence time but not the survival time. PMID- 12749252 TI - Radiofrequency ablation of hepatocellular carcinoma in cirrhotic patients. AB - BACKGROUND/AIMS: Radiofrequency ablation is useful for local destruction of primary and secondary hepatic tumors. However there is little information about long-term outcome for patients with hepatocellular carcinoma and cirrhosis. The aims of this study included the analysis of efficacy in local control of tumors, patterns of recurrence and survival rate related to severity of cirrhosis and tumor features. METHODOLOGY: From January 1998 to January 2001 we ablated 65 hepatocellular carcinomas in 53 patients with cirrhosis. Cirrhosis was related to HCV, HBV and alcohol intake in 32, 4 and 17 patients, respectively. According to Child-Pugh classification 24 and 29 patients belong to A and B class, respectively. Patients in Child-Pugh C class were excluded from the study. The mean diameter of lesions was 4 cm (range: 1-7 cm). In all cases radiofrequency ablation was performed with the percutaneous approach under ultrasound guidance. Treatment efficacy was estimated with dual phase computed tomography and alpha fetoprotein dosage. RESULTS: Complete necrosis rate after first treatment was 90.9, 74.4 and 36.4% in hepatocellular carcinoma smaller than 3 cm, between 3 and 5 cm and larger than 5 cm, respectively (p = 0.01). After multiple treatment complete necrosis was achieved in 100, 93 and 63.6% (p = 0.01). The mean number of treatments was 1.5 (range: 1-4) per patients. Four lesions of 58 (6.8%) developed local recurrence during a mean follow-up of 18 months. No local recurrence was observed in tumors smaller than 4 cm. New tumors were observed in 15 patients (28.3%). There were no treatment-related deaths and 11 patients (20.7%) suffered minor complications. The survival rate of patients after treatment was 87, 63 and 45% at 12, 24 and 36 months, respectively. Survival was significantly related to Child-Pugh class (p = 0.01). CONCLUSIONS: Radiofrequency ablation is a safe and effective treatment of hepatocellular carcinoma smaller than 5 cm with complete necrosis in most lesions. No local recurrences were observed in lesions smaller than 4 cm. After 3 years survival was 83% in Child Pugh A cirrhotic patients and 31% in Child-Pugh B patients. PMID- 12749254 TI - Hepatic resection for hepatocellular carcinoma in severely cirrhotic livers. AB - BACKGROUND/AIMS: Liver resection for hepatocellular carcinoma is typically avoided in patients with severe cirrhosis, as this may predispose patients to pronounced surgical risks and worse long-term outcomes. This study examined our results of hepatectomy for hepatocellular carcinoma in patients with different degrees of impaired liver function. METHODOLOGY: A retrospective comparison of background characteristics and operative results in patients with resected hepatocellular carcinoma was performed, with patients classified as having either severely impaired liver function (High Group, preoperative indocyanine green 15 minute retention rate > or = 30%; n = 22) or normal liver function (Low Group, indocyanine green 15-minute retention rate < or = 10%; n = 37). RESULTS: The preoperative liver function of those in the High Group was significantly worse than those in the Low Group. Despite similar tumor diameters between groups, the amount of liver resected in the High Group was significantly smaller than in the Low Group, primarily because of more limited resections with narrow margins in the former. However, patients in the High Group had similar recurrence and survival rates when compared to those in the Low Group whose resections included wide surgical margins. In analyzing those that did or did not undergo transcatheter arterial chemoembolization prior to resection, a better disease free and actuarial survival rate was observed in the High Group with preoperative transcatheter arterial chemoembolization compared to those without transcatheter arterial chemoembolization or those in the Low Group with transcatheter arterial chemoembolization. CONCLUSIONS: For patients with limited liver function reserve, such as those in the High Group, resection of hepatocellular carcinoma with preservation of liver parenchyma may take priority over a wide resection margin. In addition, we suggest the use of preoperative transcatheter arterial chemoembolization in selected patients with severe cirrhosis undergoing resection of hepatocellular carcinoma. PMID- 12749253 TI - Postoperative evaluation of C-tube drainage after hepatectomy. AB - BACKGROUND/AIMS: In spite of recent advances in liver surgery, biliary complications remain a common cause of major morbidity after hepatectomy. METHODOLOGY: We studied the postoperative evaluation of 28 hepatectomied patients with transcystic duct tube (C-tube) drainage (C-group), compared with 38 hepatectomied patients without C-tubes (NC-group), in terms of preoperative clinical profiles of patients, intraoperative findings and procedures, postoperative management and bile leakage, daily output of bile, liver function and postoperative infections. RESULTS: There were no significant differences in the preoperative clinical profiles of patients and postoperative management between the two groups. In intraoperative findings and procedures, the tumor size, weight of the resected liver, operation time and operative blood loss were higher in the C-group than those in the NC-group. Therefore, the operative procedure was more serious in the C-group than that in the NC-group. However, bile leakage was observed in only one of 28 patients (3.6%) in the C-group and 10 of 38 patients (26.3%) in the NC-group, that is, bile leakage occurred less frequently in the C-group than in the NC-group. The daily output of bile in the C group was thought to be enough to decompress the biliary tree. In liver function, aspartate aminotransferase and alanine aminotransferase had lower levels in the C group than those in the NC-group with bile leakage. White blood cell count, C reactive protein and body temperature were closer to the normal range in the C group than those in the NC-group with bile leakage. CONCLUSIONS: C-tube drainage after hepatectomy is thought to be useful for decreasing postoperative complications, especially bile leakage. PMID- 12749255 TI - Effects of marker for hepatic fibrosis and viral status on recurrence after resection of hepatitis B virus-related hepatocellular carcinoma. AB - BACKGROUND/AIMS: We investigated the relationship between the serum concentration of type IV collagen 7s domain (7s collagen) and viral status and the effects of these factors on recurrence after resection of hepatitis B virus-related hepatocellular carcinoma. METHODOLOGY: Serum concentration of 7s collagen was measured in 39 patients who underwent liver resection for hepatitis B virus related hepatocellular carcinoma. The clinicopathologic findings and tumor-free survival rate were compared between patients with a low serum concentration of 7s collagen (< 7.0 ng/mL, group 1) and those with a high serum concentration of 7s collagen (> or = 7.0 ng/mL, group 2). RESULTS: Aspartate aminotransferase activity and the proportion of patients with a high viral load and cirrhosis were significantly higher in group 2 than in group 1. The tumor-free survival rate was significantly lower in group 2 than in group 1 (P = 0.0050). The survival rate was significantly lower in patients with a high viral load than in patients with a low viral load (P = 0.0006). CONCLUSIONS: A high serum concentration of 7s collagen was closely related with a high viral load of hepatitis B virus. A high serum concentration of 7s collagen and a high viral load of hepatitis B virus are risk factors for recurrence after resection of hepatitis B virus-related hepatocellular carcinoma. PMID- 12749256 TI - The long-term efficacy of combined chemoembolization and local irradiation in the treatment of patients with large hepatocellular carcinoma. AB - BACKGROUND/AIMS: To investigate the effects and toxicity of combined transcatheter arterial chemoembolization and local radiotherapy for patients with unresectable hepatocellular carcinoma. METHODOLOGY: From June 1994 to June 1999, 28 patients with unresectable hepatocellular carcinoma (the largest dimension of tumor ranged from 3 to 10 cm) were treated with transcatheter arterial chemoembolization followed by local irradiation. Response was assessed by computed tomography scan and ultrasound, together with serum alpha-fetoprotein level in cases with increased baseline values. Complications associated with treatment were monitored. Cumulative survival was determined by Kaplan-Meier method from the first day of treatment. RESULTS: An objective response (reduction of tumor area greater than 50%) was achieved in 57.0% of the cases. In 63.0% of the cases with increased alpha-fetoprotein values, alpha-fetoprotein level underwent a marked reduction (more than 50%). The cumulative survival rates at 1, 3, and 5 years were 81.9%, 42.4%, and 21.2%, respectively (median survival, 31 months, 95% CI: 27.8-34.2 months). The combination therapy was generally well tolerated. CONCLUSIONS: The results suggest that combined transcatheter arterial chemoembolization with local radiotherapy is an effective and safe treatment for unresectable hepatocellular carcinoma. Further study is required to compare the therapeutic efficacy of this regimen to transcatheter arterial chemoembolization or radiotherapy alone. PMID- 12749257 TI - The effect of beta-blocker on intractable ascites in cirrhotic patients undergoing hepatectomy for hepatocellular carcinoma. AB - BACKGROUND/AIMS: Intractable ascites is one of the serious complications after hepatectomy. Only little is known about their effect on postoperative ascites in patients with liver cirrhosis although beta-blockers have been used for cirrhotic complications including ascites. METHODOLOGY: Here, we report five cases of intractable ascites after hepatectomy, which were treated by propranolol (1 mg/kg/body). RESULTS: In three patients, plasma renin activity and aldosterone concentrations were markedly increased before propranolol administration, but fell to normal levels thereafter. Ascites subsided in all subjects except one, who developed cardiac dysfunction. CONCLUSIONS: Beta-blockers might be a promising drug for intractable ascites in cirrhotic patients undergoing hepatectomy. PMID- 12749258 TI - Infected hepatic cyst. AB - We describe an unusual case involving an infected hepatic cyst. An 88-year-old woman presented with acute onset of right upper quadrant abdominal pain, mild left lower abdominal pain, diarrhea, and fever. On admission, computed tomography revealed multiple hepatic cysts including an 8-cm cyst located in the left medial segment of the liver, which demonstrated a thickened wall enhanced with contrast media. Ultrasonography showed an 8-cm hypoechoic lesion which differed in appearance from the other, anechoic hepatic cysts. The serum concentration of C reactive protein was 29.8 mg/dL; white blood cell count, 12,800/microL; CA19-9, 96 U/mL; and CEA, 2.2 ng/mL. Diagnosis of infected hepatic cyst was made by percutaneous transhepatic drainage of the cyst. Milky fluid was obtained and the patient's right upper quadrant abdominal pain resolved after drainage. The cyst fluid CA19-9 concentration was 18,000 U/mL. Cytology of the cyst fluid was negative. Serum CA19-9 (41 U/mL) and CEA (1.8 ng/mL) concentrations were improved 1 week after drainage. Escherichia coli was cultured from the drainage fluid. The patient was discharged 27 days after admission. Percutaneous transhepatic drainage is effective in the treatment of infected hepatic cysts. PMID- 12749259 TI - Transplantation of a ninety-three-year-old donor liver. Case report. AB - Advanced donor age has been considered a risk factor for the use of organs for transplantation. We report the case of an orthotopic liver transplant performed using a 93-year-old donor. The donor had been admitted to the intensive care unit 3 days before due to cerebral hemorrhage. History, viral serology, liver function tests and hemodynamics were normal. At laparotomy, the liver appeared macroscopically normal; histology showed mild parenchymal congestion and focal signs of steatosis (less than 10%). The liver was therefore procured and transplanted into a 52-year-old recipient with alcoholic, post-hepatitis C. cirrhosis. Cold and warm ischemia times were 8 hours 20 min and 67 min, respectively. Production of bile was observed after reperfusion. Six months post transplantation there is clinical and histological evidence of hepatitis C virus recurrence. Nevertheless, the patient enjoys an acceptable quality of life. Even very old donor livers can be used for transplantation, although it is still debatable whether hepatitis C virus-positive patients are good recipients of such livers. PMID- 12749260 TI - Early medical treatment is life saving in acute Budd-Chiari due to polycythemia vera. AB - Here we describe two women with polycythemia vera presented with fulminant hepatic failure due to acute Budd-Chiari syndrome. Both had a history of severe abdominal pain and distention of short duration. Clinical and laboratory findings showed fulminant hepatic failure due to acute Budd-Chiari syndrome. Diagnosis was confirmed with abdominal ultrasonography and Doppler ultrasonography showing ascites, hepatomegaly, portal hypertension and total occlusion of hepatic veins. Complete blood count and other clinical findings were compatible with polycythemia vera in both patients. Patients were treated successfully with early administration of continuous heparin infusion, repeated phlebotomies and hydroxyurea. We emphasize here early diagnosis and effective treatment in such fulminant cases may be life saving. PMID- 12749261 TI - Complete recovery from fulminant hepatic failure with severe coma by living donor liver transplantation. AB - In Japan, living donor liver transplantation has been established as a therapeutic strategy for the rescue of terminal liver disease, including fulminant hepatic failure that shows no signs of recovery. We performed living donor liver transplantation for a subacute type fulminant hepatic failure patient, who had developed a hepatic coma of grade V (no right reflex, no response to pain stimuli). The electroencephalogram indicated almost flat waves. However, cranial computed tomography revealed that brain edema was not severe in this case. The recipient did not have hepatitis virus and had not taken medication that had been determined to cause hepatitis. The recipient was a 12 year-old boy, 165.5 cm in height and 45.5 kg in weight. The donor was his mother, who was 42 years old; her blood type, type B, was identical to that of the boy. The mother's right hepatic lobe was transplanted to her son (the recipient). The post-transplantation condition of recipient was quite excellent. He recovered consciousness 3 days after liver transplantation, and rapidly attained normal hepatic function. The donor was discharged on the 20th postoperative day without any problems. The recipient was discharged on the 79th postoperative day without any neurological deficits. This case suggests that deep coma without electroencephalogram waves may not be a contraindication for living donor liver transplantation in fulminant hepatic failure patients, if the brain edema is not severe. PMID- 12749262 TI - Large primary liver cancer treated by chemoembolization combined with radiotherapy successfully. AB - For unresectable primary liver cancer, transcatheter arterial chemoembolization is the most effective method in reducing the size of tumor and become the most popular forms in Asia, but, it is not a curative approach and additional therapy is needed to kill the residual tumor cells. In this paper, we report a case in which a patient with a single very large liver cancer has been successfully treated by chemoembolization combined with radiotherapy. The results suggest that this is a very promising therapeutic approach for single large primary liver cancer. PMID- 12749263 TI - Surgical repair of a liver injury in a patient: accompanied with tricuspid regurgitation. AB - A 42-year-old man with acute myocarditis developed a right lung abscess and tricuspid regurgitation with valvular vegetations. A chest tube was inserted from the 8th intercostal space on the anterior axillary line to drain acute thoracic empyema caused by rupture of the lung abscess. Six hours after the insertion, the patient went into shock. Since abdominal ultrasonography showed intraabdominal bleeding, an urgent laparotomy was performed. A large amount of blood was present in the peritoneal cavity. Pulsatile bleeding from the hepatic laceration and penetration of diaphragm were observed. Mattress sutures were used to close the hepatic laceration and the rent in the right hemi-diaphragm. Because the patient was hemodynamically stable postoperatively tricuspid valvular replacement was performed 5 days after the laparotomy. Four months after valve replacement, the patient is in good condition without any complaints. PMID- 12749264 TI - Acute intraoperative hemolysis and hemoglobinuria during radiofrequency ablation of hepatocellular carcinoma. AB - Radiofrequency ablation is a safe and effective treatment for small primary or secondary liver tumors. Development of new probes has allowed the use of radiofrequency ablation for ablation of tumors > 5 cm in the liver. We present a case of acute intravascular hemolysis and hemoglobinuria during radiofrequency ablation of a large 8-cm hepatocellular carcinoma via laparotomy. The hemolysis was recognized during the operation by change of urine color, and prompt management was initiated to prevent acute renal failure. Literature search revealed that radiofrequency ablation can induce hemolysis in experimental setting, but this is the first clinical report of acute hemolysis during radiofrequency ablation for liver tumors. PMID- 12749265 TI - Tube cholangio-jejunal bypass in unresectable hepatic hilar carcinoma for a better quality of life. AB - A 76-year-old male patient with jaundice was diagnosed as having hepatic hilar cholangiocarcinoma. The patient underwent percutaneous transhepatic biliary drainage of the left and the right intrahepatic bile duct. He could not have a percutaneous transhepatic endoprosthesis placed because it was impossible to pass the guidewire through the stenotic portion of the lesion. The patient was operated, but the tumor was considered to be unresectable. Along the intrahepatic routes formed by the preexisting two percutaneous transhepatic biliary drainage tubes, silicon tubes were inserted. Through the lumen of a long jejunal limb, Roux-en-Y, the tubes with five to six side holes were passed in the distal direction and drawn out from the lumen of the jejunal limb and passed through the abdominal wall to outside. The tubes were occluded and buried in the subcutaneous space after a few clamping tests. He died of liver failure in his house four months after the operation without any symptoms of jaundice, fever or of obstruction of the tubes. PMID- 12749266 TI - Conformal radiation therapy for liver metastasis of esophageal carcinoma. AB - Recently, aggressive hepatectomies or hepatic arterial infusion chemotherapy for liver metastasis from gastric or colorectal carcinoma have been performed, and the number of successful studies of liver metastasis have increased. However, there have been few successful cases of liver metastasis from esophageal carcinoma by surgery or chemotherapy. Herein, we show the benefits of radiation therapy for the treatment of liver metastasis from esophageal carcinoma. A 60 year-old woman with a 5-cm solitary liver metastasis from esophageal squamous cell carcinoma was treated with radiation therapy. The treated volume was encompassed by the anteroposterior and right lateral opposing fields, shaped by a multileaf collimator. The daily fraction size was 1.8 Gy, 5 days per week, for a total dose of 54 Gy. During the course of treatment, the patient did not experience any complications. After radiotherapy, abdominal computed tomography showed that the enhanced solid tumor had changed to a very low-density mass lesion with a clear margin, and the size was decreasing gradually between the 6 months. Radiotherapy could be a treatment of choice in patients with liver metastasis from esophageal squamous cell carcinoma. PMID- 12749267 TI - Peculiar glucagon processing in the intestine is the genesis of the paradoxical rise of serum pancreatic glucagon in patients after total pancreatectomy. AB - BACKGROUND/AIMS: Although glucagon has been detected even in the serum of totally pancreatectomized patients and the origin was suggested to be the intestine, the kinetics of glucagon are not well known after pancreatectomy. In the present study, we investigated the kinetics of glucagon and glucagon-related peptides in pancreatectomy patients and discuss the glucagon processes. METHODOLOGY: Ten patients who had undergone total pancreatoduodenectomy reconstruction using Billroth II type procedures (group PX) and 12 normal subjects (group C) were also enrolled in this study. All patients received a 75-g oral glucose tolerance test in the early morning fasting state. Serum glucagon levels were assessed using the glucagon specific C-terminal (immunoreactive glucagon: IRG) and nonspecific N terminal (glucagon-like immunoreactivity: GLI) radioimmunoassays. The molecular forms of these glucagon-related peptides were also estimated using the gel filtration chromatography method before and after the oral glucose load. RESULTS: After the glucose load, serum GLIs were increased significantly in group PX suggesting that these were affected by the alimentary tract reconstructions. Serum IRGs were significantly increased in group PX, but decreased in group C after oral glucose load suggesting that these paradoxical increased responses in group PX might be associated with the insulin secretion deficiencies, but not associated with the alimentary tract reconstruction. CONCLUSIONS: The paradoxical rise in IRGs based on the findings of gel filtration chromatography in group PX were possibly due to the generated peculiar glicentin-like peptide from the glucagon precursor, preproglucagon, after total pancreatectomy, which might be processed in intestines in association with the insulin deficiencies. PMID- 12749268 TI - P53 mutation but not p16/MTS1 mutation occurs in intraductal papillary mucinous tumors of the pancreas. AB - BACKGROUND/AIMS: Intraductal papillary mucinous tumors of the pancreas are rare lesions, which typically show a benign clinical course. However, some of these tumors have a malignant nature and grow in an invasive manner. The purpose of the study was to determine the prevalence of p53-, p16/MTS1- and K-ras mutations in benign and malignant intraductal papillary mucinous tumors with intent to value their importance for tumor progression. METHODOLOGY: Thirteen different archival tumor specimens were obtained at the Department of Pathology, University of Ulm. Three cases showed an invasive component of the tumor. Genomic DNA was extracted after laser capture microdissection of tumor cells from paraffin-embedded tissue sections. The corresponding sequences of p53 (exon 5, 6, 7, 8) and p16/MTS1 (exon 2) were amplified by polymerase chain reaction and subjected to single strand conformation polymorphism analysis. Codon 12 of K-ras was analyzed by the enrichment polymerase chain reaction-restriction fragment length polymorphism method. Positive samples were further investigated by sequencing. RESULTS: K-ras mutations occurred in benign and malignant intraductal papillary mucinous tumors (4/13), whereas an alteration of the coding p53 gene sequence could only be detected in the intraductal and invasive component of one malignant tumor. None of the tissue specimens revealed mutations in exon 2 of p16/MTS1. CONCLUSIONS: In contrast to K-ras mutations, alterations in the p53 gene may characterize ductal papillary mucinous carcinomas, which could be of major interest for their early diagnosis. The lack of mutations in the p16/MTS1 gene suggests that other genes may be involved in the formation of intraductal papillary mucinous neoplasias. PMID- 12749269 TI - Pancreaticogastrostomy: unreliable long-term pancreatic duct patency. AB - BACKGROUND/AIMS: Long-term patency of pancreaticogastrostomy remains unclear. The purpose of this study is to clarify the long-term patency of pancreaticogastrostomy based on change in the main pancreatic duct diameter. METHODOLOGY: Seventeen of 33 patients who underwent pancreaticogastrostomy in our institution were divided into 2 groups according to the preoperative diameter of main pancreatic duct: the non-dilated group (phi main pancreatic duct < or = 4 mm, 11 patients) and the dilated group (phi main pancreatic duct > or = 5 mm, 6 patients). Clinical and radiological parameters were assessed by a postoperative comparison between the 2 groups. RESULTS: phi main pancreatic duct after operation was dilated in all 11 patients in the non-dilated group, and the difference was significant (p = 0.0003, mean dilatation ratio = 2.6) when compared with phi main pancreatic duct before operation. In contrast, the mean phi main pancreatic duct decreased after operation (mean dilatation ratio = 0.9) in the dilated group. Clinical symptoms (5 patients), impaired endocrine and exocrine pancreatic function (4 and 3 patients, respectively), elevated serum amylase level (6 patients), and pancreatic parenchymal changes on radiological examinations (5 patients) were observed only in the non-dilated group. CONCLUSIONS: Degree of dilatation of main pancreatic duct seems to correspond to likelihood of anastomotic stenosis of pancreaticogastrostomy. Pancreaticogastrostomy has less reliability in terms of the long-term patency in cases having non-dilated main pancreatic duct. PMID- 12749270 TI - Regional chemotherapy for advanced pancreatic carcinoma. AB - BACKGROUND/AIMS: Since 1990, 14 patients with advanced pancreatic cancer were treated by regional chemotherapy. Five patients had locally advanced unresectable cancer and 9 had locally advanced disease with liver metastasis. METHODOLOGY: Fourteen patients underwent laparotomy, splenic artery catheterization and received a mean therapy of 3.8 cycles and 3 patients had completed 6 cycles of chemotherapy. Every cycle given monthly consisted of 5-fluorouracil 600 mg/m2 3 days, mitomycin 10 mg/m2 1 day, and cisplatinum 60 mg/m2 1 day. Tumor response was evaluated on the basis of imaging methods, tumor markers and life quality marker pain relief. RESULTS: Four patients had stabilization, 4 had partial response, 5 had progression and 1 had complete response. Median survival was 8 months for the whole group. Palliation for pain was successful and 71.4% of the patients had pain palliation. One patient had complete, 1 patient had partial, 8 patients had stable pain relief. Four patients had poor response to treatment and had progressive pain. Side effects, mainly leukopenia and thrombocytopenia, occurred in 5 patients and responded to medical measures. Mild to moderate nausea and vomiting was common and they responded well to antiemetic treatment. CONCLUSIONS: Although the objective response rate of regional chemotherapy for overall survival is low, in an individual patient it may produce an adequate response and acceptable toxicity so that the patient experiences overall improvement in symptoms, and rarely as in one case in our group may be cured by this method. PMID- 12749271 TI - Results of treating severe acute pancreatitis with gabexate is associated with neutrophil apoptosis activity. AB - BACKGROUND/AIMS: A delay in polymorphonuclear neutrophil apoptosis has been implicated in the pathogenesis of systemic inflammatory reactions in certain conditions. Gabexate mesilate has been proven effective in the treatment of severe acute pancreatitis with organ dysfunction. In this study, we attempted to answer the questions of whether neutrophil apoptosis is associated with the conditions of various major organs in patients with severe acute pancreatitis and receiving gabexate. METHODOLOGY: A total of 45 patients were included in this study. We divide the patients into two groups. Group A included patients with > or = 2 complications after one-week treatment (n = 31), and Group B included patients with < 2 complications after one-week treatment (n = 14). Serum interleukin-6 and interleukin-8 was detected at day 1, 3, and 7 of treatment using enzyme-linked immunosorbent assay kits. The neutrophil CD18 expression and apoptosis activity were evaluated flowcytometrically at day 1, 3, and 7 of treatment. RESULTS: At day 7 of treatment, interleukin-6 levels were significantly higher in Group B while interleukin-8 levels were not different. The neutrophil CD18 expression was significantly higher and delayed ex vivo apoptosis was significantly lower in the group B than that of group A at day 7 of treatment. CONCLUSIONS: In patients of severe acute pancreatitis with organ dysfunction and receiving gabexate treatment, neutrophil apoptosis is associated with the severity of organ dysfunction. PMID- 12749272 TI - Virtual pancreatoscopy of pancreatic cancer. AB - BACKGROUND/AIMS: Virtual endoscopy is a new method of diagnosis using computer processing of three-dimensional images data sets. However, there are few reports about the clinical application of virtual endoscopy for the pancreas. In this study, we evaluated the feasibility of surface-rendered magnetic resonance virtual endoscopy for pancreatic cancer. METHODOLOGY: Twenty-six cases of pancreatic cancer were studied. Fifteen patients had pancreatic head cancer, 7 had pancreatic body cancer, and 4 had pancreatic tail cancer. Twelve patients underwent surgical resection of the pancreas. Magnetic resonance imaging data were acquired with a 1.5-T clinical imager (Signal.5; GE Medical Systems, USA). We used a multislab single-shot fast spin-echo sequence. Section thickness was between 2 and 3 mm in the coronal plane. Three-dimensional reconstructed images and virtual endoscopic images were generated with Advantage Windows by GE. RESULTS: Virtual endoscopic images could be generated in 20 patients with pancreatic cancer (76.9%). In these cases, we were able to observe the inner surface of the pancreatic duct and the stricture from not only the pancreatic head but also the pancreatic tail. Clear virtual images could not be generated in 6 cases. We were able to divide the 20 cases in which images could be generated into groups according to the appearance of the stricture. The edge of the stricture appeared to be protruding in 4 cases (15.4%), and appeared to be polygonal in 13 cases (50.0%). In 3 cases, we recognized the existence of a stricture, but the detail of the stricture was unclear. CONCLUSIONS: Virtual endoscopy caused minimal discomfort compared to real endoscopic examination, and it can access cystic lesions and the pancreatic duct behind the stricture. It is concluded that virtual endoscopy for pancreatic cancer has potential clinical utility. PMID- 12749273 TI - Resection of cancer of the body and tail of the pancreas. AB - BACKGROUND/AIMS: To report our results of resection of cancer in the body and tail of the pancreas and review the literature. METHODOLOGY: Thirteen patients with a median age of 62 years with cancer of the body and/or tail of the pancreas. The diagnosis was made by ultrasonography, computed tomography, endoscopic retrograde cholangiopancreatography and angiography. Eleven patients had distal or subtotal pancreatectomy and two patients total pancreatectomy. The surgical procedure included extensive dissection of lymph nodes and the connective tissue in the peripancreatic region. Main outcome measures were postoperative morbidity and mortality, median and 5-year survival rates. RESULTS: Ten of the resections were considered to be curative. Postoperative complications occurred in seven patients and one patient died in the postoperative period. The median survival time of operative survivors was 392 days. Two patients survived five years, and one was alive ten years after surgery. Eight patients died of recurrence. CONCLUSIONS: Long-term survival may be achieved in a quarter of the resectable patients. PMID- 12749275 TI - Intrahepatic hemorrhage and subcapsular hematoma developing in acute pancreatitis. AB - Pancreatitis with hemorrhagic complication is a rarely reported situation. It is always devastating and possibly fatal if massive bleeding occurs. We report a case of acute pancreatitis associated with renal failure who developed intrahepatic hemorrhage and subcapsular hematoma. Bleeding was successfully controlled by emergent embolization via proper hepatic artery. The patient recovered uneventfully later. PMID- 12749274 TI - A phase II study of docetaxel-irinotecan combination in advanced pancreatic cancer. AB - BACKGROUND/AIMS: Chemotherapy provides dismal results in advanced pancreatic cancer patients, even when new compounds, such as gemcitabine, are used. Phase I studies of single-drug therapy with docetaxel or irinotecan suggested a response rate of about 15% in these patients. We report here a phase II study of docetaxel irinotecan combination in advanced pancreatic cancer patients. METHODOLOGY: Docetaxel 60 mg/m2 was given in combination with irinotecan 250 mg/m2 every 3 weeks. Prednisolone premedication and anti-HT3 drugs were systematically administered. Hematopoietic growth factors were given in case of febrile neutropenia or grade 4 neutropenia at the previous cycle. Endpoints were response rate, progression-free survival, and tolerance. RESULTS: Twenty-seven patients were enrolled, of whom 25 had metastatic disease. We observed 3 partial responses and 11 stabilizations. The median progression-free survival was 4.3 months. Myelosuppression was the main toxicity with 18% of patients experiencing a grade 3-4 event. One patient died of neglected febrile neutropenia. Gastrointestinal toxicity was well controlled. Other toxicities were mild. CONCLUSIONS: This combination has acceptable tolerance and, despite an 11% response rate, some partial responses and prolonged stabilizations were observed. The treatment induced clinical benefit in 33% of the patients. Further trials should focus on docetaxel or irinotecan, possibly used in combination with more conventional strategies (gemcitabine). PMID- 12749276 TI - Laparoscopic splenectomy for variceal bleeding with non-cirrhotic portal vein thrombosis: a case report. AB - A 57-year-old man was referred to our hospital for treatment of refractory gastric bleeding from gastric varices secondary to portal vein thrombosis. The patient's liver function tests and coagulation profile were normal. The venous phase of the superior mesenteric arteriogram, on the other hand, showed superior mesenteric vein-portal vein occlusion with surrounding hepatopetal variceal collaterals. The venous phase of the splenic arteriogram additionally showed splenic vein occlusion and collateral vessels from the gastric and retroperitoneal regions flowing into a portal cavernous transformation. Gastroscopy confirmed that the patient had gastric varices in the cardia. We performed laparoscopic splenectomy to treat refractory gastric bleeding from varices and symptomatic hypersplenism. The postoperative course was uneventful; the patient's gastric varices were less prominent on follow-up gastroscopy and the hematologic profile returned to normal. Extrahepatic portal vein thrombosis is the leading cause of variceal hemorrhage in patients with healthy livers. There is a consensus in the literature that splenectomy alone is of minimal value in preventing variceal bleeding in portal vein thrombosis. Splenectomy is, however, indicated in cases in which the patient has hepatopetal collaterals from the mesenteric vein system and whose hemorrhagic gastric varices are related to splenic vein thrombosis as in our case. PMID- 12749277 TI - Inhibition of peptic ulcer relapse by ranitidine and ecabet independently of eradication of Helicobacter pylori: a prospective, controlled study versus ranitidine. AB - BACKGROUND/AIMS: The recent increase in resistant strains of Helicobacter pylori has become a serious problem. Ecabet is a novel anti-ulcer agent that acts directly on the gastric mucosa, has bactericidal activity, and inhibits adhesion of Helicobacter pylori to the gastric mucosa. These actions result from inhibition of urease and ATPase in Helicobacter pylori, a mechanism distinct from that of antibiotics. METHODOLOGY: Sixty-three patients positive for Helicobacter pylori who had been cured of gastric ulcers and duodenal ulcers were randomly assigned to receive maintenance therapy with ranitidine alone or a combination of ranitidine and ecabet. Ulcer relapse was studied in these patients. RESULTS: The cumulative relapse rates in the ranitidine group and the ecabet plus ranitidine group were respectively, 29.6% and 4.4% after 1 year of treatment and 66.1% and 13.0%, after 2 years. These differences were significant (p = 0.006). Multivariate analysis of factors potentially related to relapse showed that outcome was significantly related only to treatment (p = 0.020) and not to other characteristics, such as age, diagnosis, or sex. CONCLUSIONS: We conclude that maintenance therapy with a combination of ranitidine and ecabet prevents ulcer relapse in Helicobacter pylori-positive patients. Controlled studies comparing ulcer relapse rates between eradication treatment and maintenance therapy with ranitidine and ecabet are awaited. PMID- 12749278 TI - The effect of different frequency electrical acu-stimulation on gastric myoelectrical activity in healthy subjects. AB - BACKGROUND/AIMS: Acupuncture has been used to treat gastrointestinal symptoms in oriental countries for many years. The underlying mechanism is still not fully understood. METHODOLOGY: Fifteen healthy male volunteers were enrolled into this study. A cutaneous electrogastrography recording was obtained. Two frequencies (2 Hz and 100 Hz) of electrical stimulation were applied to Zusnali or Shousanli point in different sessions. Another non-acu-point (2 cm lateral to Zusanli) without electrical stimulation was used for a control study. RESULTS: There was a significant increase in the percentage of normal frequency during 2 Hz of electrical stimulation on Zusnali (baseline vs. acupuncture, 82.49 +/- 12.87% vs. 93.18 +/- 8.40%, p < 0.01). The percentage of normal frequency did not change significantly, during or after acupuncture, with 100 Hz of electrical stimulation on Zusnali, or 2 Hz and 100 Hz of electrical stimulation on Shousanli. In addition, the percentage of tachygastric frequency and power ratio also changed significantly during 2 Hz of electrical stimulation on Zusnali. However, the change in the percentage of bradygastria and dominant frequency was not statistically significant among all five study sessions during each stage. CONCLUSIONS: The results of this study demonstrated that electrical stimulation, with a frequency of 2 Hz, on Zusanli might enhance the regularity of gastric myoelectrical activity. The effect of acupuncture on gastric myoelectrical activity was acu-point-specific, and that effect was observed with 2 Hz but not 100 Hz electrical stimulation. PMID- 12749279 TI - Gastric remnant cancer compared with primary proximal gastric cancer. AB - BACKGROUND/AIMS: Recently, detections of early-stage gastric remnant cancer and small proximal gastric cancer are increasing. The aim of this study was to compare pathologic and prognostic data of gastric remnant cancer with those of primary proximal gastric cancer including upper gastric cancer based on a recent 15-year experience at a single institute in Japan. METHODOLOGY: Among 698 patients who underwent gastrectomy for cancer between 1984 and 1998, 15 (2.1%) were patients with gastric remnant cancer. During the same period, 139 patients underwent primary gastrectomy for proximal gastric cancer which included 71 with upper gastric cancer confined to the upper one-third of the stomach. Clinicopathologic findings of gastric remnant cancer were compared with those of proximal gastric cancer. RESULTS: Of 15 gastric remnant cancers, 8 (53%) were stage I tumors. Although gastric remnant cancer and proximal gastric cancer was not different in several clinicopathologic factors, gastric remnant cancer and upper gastric cancer confined to the upper one-third of the stomach was different with regard to the frequency of tumor size > or = 4 cm (60% vs. 32%, p < 0.05), poorly differentiated type (67% vs. 38%, p < 0.05), serosal invasion (40% vs. 11%, p < 0.01), lymph node metastasis (47% vs. 20%, p < 0.05), stage III or IV disease (47% vs. 10%, p < 0.01), and noncurative gastrectomy (20% vs. 1%, p < 0.01). The 5-year survival rate of gastric remnant cancer (69%) was higher than that of proximal gastric cancer (57%) and lower than that of upper gastric cancer (81%), although the differences were not statistically significant. CONCLUSIONS: In our recent series, a half of gastric remnant cancers are stage I tumors. Although gastric remnant cancers are similar to proximal gastric cancers, they are more advanced and their surgical results are less satisfactory when compared with upper gastric cancers confined to the upper one-third of the stomach. PMID- 12749281 TI - Is dental care really that expensive? PMID- 12749280 TI - Premalignant lesions and conditions for gastric adenocarcinoma: diagnosis, management and surveillance guidelines. AB - Despite universal trends of declining mortality rates, gastric cancer remains a major health problem. The advent of endoscopy has made a major impact in the diagnosis and management of early gastric cancer as well as of the premalignant lesions of the stomach. Nevertheless the disagreement between Japanese and Western histopathologists regarding the nomenclatures of the gastric precancerous lesions resulted in different recommendations and therapeutic guide lines. On this background we review the current concepts related to premalignant lesions, especially intestinal metaplasia and gastric dysplasia as well as the critical role of Helicobacter pylori in the evolution of these lesions. Additionally, this extensive review deals with the majority of precancerous conditions related to gastric adenocarcinoma and provides surveillance and therapeutic guidelines. PMID- 12749282 TI - "Grin and bear it"--the frustrated denture wearer response to ethical dilemma #42. PMID- 12749283 TI - Oral and maxillofacial pathology case of the month. Gingival cyst of the adult. PMID- 12749284 TI - [Outcome predictors in patients with normal pressure hydrocephalus]. AB - Despite the accumulation of knowledge over the years, the postoperative results of shunt implantation in patients with normal pressure hydrocephalus (NPH) have shown little improvement. This means that reliable predictors of the course of the disease need to be identified. In a prospective study carried out between 1982 and 2000 we re-examined 155 (78%) of 200 NPH patients treated by shunt implantation, 7 months after their operation. On the basis of the results of the intrathecal infusion test NPH was graded early stage (no brain atrophy) or late stage (brain atrophy). We looked at the following factors as possible predictors: patient's age, disease duration, idiopathic or secondary aetiology, clinical signs such as gait ataxia, dementia and urinary incontinence, results of spinal tap, valve type and valve infection, and resistance to cerebral spinal fluid outflow and postoperative changes in ventricular size. As a measure for outcome we used the NPH recovery rate, and the Pearson chi-square test for statistical evaluation. 80 patients with early stage NPH, a history < 1 year, absence of dementia and an implanted Miethke dual-switch valve proved to be significant predictors of a positive outcome. Outflow resistance proved to have only minimal impact on outcome. The 75 patients with late-stage NPH had better outcome when dementia was absent, outflow resistance was > 20 mmHgmin/ml, the CSF tap test was positive, and a Miethke dual-switch valve was implanted. PMID- 12749285 TI - Intraocular microablation of choroidal tissue by a 308 nm AIDA excimer laser for RPE-transplantation in patients with age-related macular degeneration. AB - Age-related macular degeneration (AMD) is the leading cause of legal blindness in the western nations beyond 50 years of age. The most frequent cause for severe visual loss is the growth of neovascular membrances from the choroid into the subretinal space. This usually results in irreversible degeneration of the overlying retina. Surgical removal of the membrane is feasible, however, usually results in functional loss of apposing retinal photoreceptors since retinal pigment epithelial (RPE) cells are removed concurrently due to their tight adherence to the neovascular complex. Therefore, various attempts have been undertaken to fill the resulting RPE cell defect with either heterologous or autologous RPE cell transplants. So far cell survival, function and subsequent visual function has been disappointing. To minimize trauma and resulting dedifferentiation harvesting in the eye and transplantation in whole sheets and without temporary removal from the eyes would be desirable. This may be achieved by isolating grafts consisting of choroid, Bruch's membrance and RPE cells from the peripheral retina and transplantation of this graft under the neurosensory retina after removal of the choroidal neovascularization. However, the choroidal component of such a graft would be expected to interfere with diffusion of metabolites to and from the retina. Therefore, outcome would be expected to be better if the choroidal tissue would be removed before translocation. In preclinical experiments we used a 308 nm UV AIDA excimer laser to microablate choroidal tissue from such a graft in human donor eyes. PMID- 12749287 TI - [Numerical study of tension and strain distribution around rat molars]. AB - A knowledge of the mechanical processes triggered in the bone and periodontal ligament (PDL) by orthodontic forces applied to a tooth is of decisive importance for an understanding of the subsequent remodelling around the tooth. To investigate these mechanical relationships, three-dimensional finite element (FE) models of the first lower molar in the rat were established. On the basis of digitized serial histological sections, these FE models were generated semi automatically. Using various simplified geometrical variations, an appropriate FE model for the analysis of the stress and strain distributions was established. The numerical analyses were carried out under a mesially directed force of 0.1 N. Stress distributions in the bone and PDL showed a similar pattern, while strains in the bone were lower than in the PDL by a factor of 10-5. The data confirm the assumption that strain patterns in the PDL may be the key stimulus of bone remodelling. PMID- 12749286 TI - [Measuring opening pressure in lumbar puncture--a new method]. AB - Determination of the opening pressure (OP) during diagnostic lumbar puncture (LP) yields additional information that may impact on treatment and prognosis in disorders affecting the central nervous system (e.g. meningitis). Established methods contain systematic errors as well as risks to the patient. We therefore present a new procedure that allows measurement of the OP by timing the flow of cerebrospinal fluid through a capillary attached to an LP needle. A resistance located between needle and capillary slows down the flow of cerebrospinal fluid so that it becomes independent of the capillary forces acting on it. The time required for the fluid to travel between two marks on the capillary (defining a given volume) can be used to calculate the flow. Since the combined resistance of needle and resistance can be calibrated, the pressure driving the flow--in this case the opening pressure--can be calculated. A simple model was used to evaluate the impact of different resistances and different needles on OP determination. The effects of cellular elements and proteins in the CSF are discussed. PMID- 12749288 TI - [Influence of the posterior tibial tendon on the medial arch of the foot: an in vitro kinetic and kinematic study]. AB - INTRODUCTION: The respective contributions of the active and passive structures of the foot to the stability of the medical arch were investigated using an in vitro kinetic and kinematic model. The effect of the tibialis posterior tendon on foot and ankle movements, and plantar pressure distribution of the foot were tested in a cadaveric human foot. METHOD: The stance phase from heel-contact to toe-off of normal walking gait and after tibialis posterior tendon rupture was simulated in eight roentenographically normal human feet (age 66 +/- 19 years, males). Ground reaction force and tibial inclination was simulated by means of a tilting angle and force-controlled translation stage. Plantar pressure was measured using a pressure-measuring platform. The force developed by the flexors and extensor muscles of the foot were simulated via cables attached to 7 force controlled hydraulic cylinders. Tibial rotation was produced by an electric servo motor, and foot movements measured with an ultrasonic analysis system. RESULTS: The model was verified against the plantar distribution and kinematics of healthy subjects measured during normal gait. Tibialis posterior deficit did not result in any detectable changes in pressure or force-time integral in the medial regions of the foot--a common sign of flat foot (pressure: midfoot 0.2 < or = 0.9; medial forefoot 0.5 < or = p < or = 0.9; hallux 0.5 < or = p < or = 0.9; force-time integral: midfoot p = 0-871; medial forefoot p = 0.632; hallux p = 0.068). Only small tendential changes in the kinematics of the talus and calcaneus were observed in dorsiflexion (0-58 sec; talus 0.1 < or = p < or = 0.6; calcaneus 0.4 < or = p < or = 0.06) and eversion (talus: 0-60 sec. 0.1 < or = p < or = 0.6; calcaneus: 37-60 sec. 0.2 < or = p < or = 0.7). CONCLUSION: The results of this in vitro study show that defective tibialis posterior alone does not produce significant changes in the kinetics or kinematics of the stance phase of normal gait. This suggests that the development of flat foot observed in degeneration of the tibialis posterior tendon occurs only after fatigue of the passive structures of the foot. PMID- 12749289 TI - [Drive principles of a triaxial physiological hip joint simulator for wear testing of hip endoprostheses]. AB - The long-term performance of hip joint replacements is a major criterion for quality assessment. Aseptic loosening as a result of wear has a considerable impact on the long-term stability of the prosthesis. Constant improvements in hip joint endoprostheses necessitate the use of hip joint simulator to evaluate and assess the suitability of newly developed materials. In the present paper, a triaxial physiological hip joint simulator is presented, and detailed information provided, on the applied control concepts, adaptation of various hard- and software systems and the accuracy of the kinematic and force parameters achievable. PMID- 12749290 TI - Cyclooxygenase-2 mediates mucin secretion from epithelial cells of lipopolysaccharide-treated canine gallbladder. AB - Biliary mucin was regarded as a major contributing factor in formation of pigment stones as well as cholesterol ones. The aim of this study was to elucidate the mechanism of biliary mucin secretion in canine gallbladder epithelial (CGBE) cells treated by lipopolysaccharides (LPS) with special reference to cyclooxygenase (COX) -2. Confluent CGBE cells were incubated with following compounds for 8, 12, and 24 hr: (1) serum-free medium, (2) serum-free medium containing LPS (100 microm/ml), (3) serum-free medium containing LPS (100 microm/ml) with NS-398 (10 microM), and (4) serum-free medium containing LPS (100 microm/ml) with indomethacin (10 microM). Mucin assay and western blots for COX-1 and COX-2 were performed. Production of PGE2, and cAMP was also measured. Mucin secretion increased with time. At 12 hr, mucin secretion increased to 200% of control (from 100 +/- 5 to 200 +/- 45%, P < 0.05). LPS treatment significantly stimulated the COX-2 expression (P < 0.05). The productions of PGE2 and cAMP were increased from 299 +/- 68 to 524 +/- 163 pg/mg (P < 0.05) and from 0.2 +/- 0.1 to 0.92 +/- 0.4 pmol/ml (P < 0.05), respectively. NS-398, which completely inhibited COX-2 expression, significantly suppressed the level of PGE2 and cAMP as well as mucin secretion (P < 0.05). Indomethacin, which partially inhibited COX-2 expression, suppressed the production of PGE2, but not cAMP and mucin secretion. In conclusion, our results suggested that the PGE2 induced by COX-2 might play a role in mucin secretion from the gallbladder epithelium through the increment of cAMP. PMID- 12749291 TI - Modified astigmatism dial diagram for locating eccentric fixation in patients with central scotoma. AB - PURPOSE: Patients with central scotoma often develop eccentric fixation on a preferred retinal locus (PRL). Identifying the PRL is one of the first steps in low vision rehabilitation training. We present our evaluation of a simple test designed to locate the eccentric fixation in eyes with central scotoma. METHODS: This was a prospective case series of consecutive patients with age-related macular degeneration and bilateral central scotoma. A numeral was added in the center of an astigmatism dial diagram. After one eye was patched, patients with central scotoma were asked to fixate the dial and describe it, then to look at the 12 o'clock position, and then around the clock. The eccentricity at which the central numeral was best seen was compared with the one determined by scanning laser ophthalmoscopy (SLO). The modified astigmatism dial test and SLO were done independently by two masked investigators. The results of the two methods were expressed in clock hour positions and were considered to be in agreement when they did not differ by more than one hour. RESULTS: Nine consecutive patients (18 eyes) with severe age-related macular degeneration and bilateral central scotoma were tested. The six women and three men ranged in age from 61 to 86 years (mean 75.8 years). The pattern test correlated with SLO findings in 12 (66%) of the 18 eyes. When considering the best-seeing eye of each patient, results showed agreement in eight (88%) out of nine cases. CONCLUSIONS: The modified astigmatism dial test appears useful for establishing the location of the eccentric fixation in the best-seeing eye of patients with bilateral central scotoma, allowing visual rehabilitation training to be started without delay. PMID- 12749292 TI - Micromechanics-based digitally controlled tunable optical beam shaper. AB - A digitally controlled programmable optical beam shaper that uses a two dimensional (2-D) small-tilt micromirror device is introduced. The optical intensity profile at any spatial position can be independently modified by use of a macropixel, incorporated with binary pulse-width modulation. An experimental proof-of-concept optical beam shaper with a commercial 2-D digital micromirror device has shown that the spatial profile of a He-Ne laser beam can be digitally manipulated. Investigation of the analog intensity control showed 256-level nonlinear degamma behavior with a measured 24.5-dB optical contrast ratio and a 10-bit spatial resolution. The performance of this tunable optical beam shaper is limited by the dimensions of the micromirror, the intermirror distance, the size of the optical beam, the number of bits used to control the micromirror, the diffraction effect, and the quality of the imaging optical system. PMID- 12749293 TI - The effect of excimer laser keratectomy on corneal glutathione-related enzymes in rabbits. AB - Glutathione related enzymes are involved in the metabolism and detoxification of cytotoxic and carcinogenic compounds as well as reactive oxygen species. Excimer laser is a very useful tool for the treatment of refractive errors and removing superficial corneal opacities. Previous studies have shown that excimer laser may initiate free radical formation in the cornea. In the present study, we evaluated the effect of excimer laser keratectomy on corneal glutathione-related enzyme activities in rabbits. Animals were divided into five groups, and all groups were compared with the controls (group 1), after epithelial scraping (group 2), transepithelial photorefractive keratectomy (PRK) (group 3), traditional PRK (group 4) and deep traditional PRK (group 5). Corneal glutathione peroxidase (GPx), glutathione S-transferase (GST) and glutathione reductase (GR) activities were measured after 24h. Corneal GPx and GR activities significantly decreased only in group 5 (p < 0.05) but GST activities significantly decreased in all groups when compared with the control group (p < 0.05). In conclusion, excimer laser inhibits the glutathione dependent defense system in the cornea, this effect becomes more prominent after high doses of excimer laser energy and antioxidants may be useful to reduce free radical mediated complications. PMID- 12749294 TI - [Born to see, referred for scanning--the ophthalmological patient in a diagnostic center]. PMID- 12749297 TI - [Caution on joining a group regarding civil rights]. PMID- 12749296 TI - [Targeted patient marketing]. PMID- 12749298 TI - Mediastinal mass with dysphagia in an elderly patient. PMID- 12749299 TI - Anthrax, botulism and tularemia in Italy. PMID- 12749300 TI - IFN-alpha: therapeutic options in HIV and HIV/HCV, HIV/HBV dual infection. AB - The antiviral effects of interferon-alpha (IFN-alpha) are well documented for the treatment of hepatitis B or C infections, whereas antiretroviral effects for treatment of HIV-1 infection have been investigated up to now only in small-sized preclinical studies. New preclinical and clinical data on IFN-alpha for HIV, HCV, HBV and especially on co-infections of HIV with HCV or HBV were presented at a symposium of the ConVir conference 2002. This short paper summarizes the presented data on IFN-alpha treatment for HIV and/or HCV or HBV infections. PMID- 12749302 TI - Low-energy extracorporeal shock-wave therapy for chronic plantar fasciitis. PMID- 12749301 TI - [Immunonutritive enteral feeding in operative intensive medicine]. PMID- 12749303 TI - [Can anesthesia be monitored by nursing personnel?]. PMID- 12749304 TI - Walking, jogging, occupational twisting, and other predictors of sciatic pain. PMID- 12749306 TI - High-velocity resistance training for increasing peak muscle power in elderly women. PMID- 12749305 TI - Courage and caring: step up to your next level of nursing excellence. PMID- 12749308 TI - Bibliography. Current world literature. Respiratory system. PMID- 12749307 TI - [Evidence based medicine and the treatment of ARDS]. PMID- 12749309 TI - Ten ways to improve information technology in the NHS. Any change in information technology in general practice requires extensive piloting. PMID- 12749310 TI - Ten ways to improve information technology in the NHS. Article states the obvious. PMID- 12749311 TI - Ten ways to improve information technology in the NHS. Current situation in the United Kingdom need not be so. PMID- 12749312 TI - Ten ways to improve information technology in the NHS. Primary care doctors need to become aware of training opportunities. PMID- 12749313 TI - ["Blackened apartment" phenomenon--practical environmental medicine approach based on a case report]. PMID- 12749314 TI - Ten ways to improve information technology in the NHS. Computers in the workplace are of great value. PMID- 12749315 TI - Intermittent peroneal mononeuropathy due to proximal tibiofibular synostosis. PMID- 12749317 TI - Closing the digital divide. Financial and technical obstacles must be eliminated. PMID- 12749318 TI - Unilateral disappearance of essential tremor after cerebral hemispheric infarct. PMID- 12749319 TI - On Johnson M, Wester K: Full remission of tardive dyskinesia following general anaesthesia. PMID- 12749320 TI - Thalamic involvement of Epstein-Barr virus encephalitis demonstrated by MRI. PMID- 12749321 TI - A case of relapsing neuromyelitis optica treated with glatiramer acetate. PMID- 12749322 TI - Abnormal cutaneous thermal thresholds in patients with restless legs syndrome. PMID- 12749323 TI - Reversible leukoencephalopathy due to vitamin B12 deficiency in an acromegalic patient. PMID- 12749324 TI - Chronic cough. Don't forget pertussis. . . PMID- 12749326 TI - Chronic cough. . . .tuberculosis . . . PMID- 12749325 TI - Non-Picks frontotemporal dementia imitating schizophrenia in a 22-year-old man. PMID- 12749327 TI - In favor of a role of lactic acid on the generation of the exercise pressor reflex. PMID- 12749329 TI - Adenosine and the exercise pressor reflex: a controversy resolved? PMID- 12749328 TI - On S. Seygert, V. Kunzmann, N. Schwertfeger, H. C. Koch, A. Faulstich: Determinants of lumbar CSF protein concentration. PMID- 12749330 TI - Chronic cough. . . .and tropical pulmonary eosinophilia. PMID- 12749332 TI - Research highlights from the literature. PMID- 12749331 TI - Neurotological findings in a family with episodic ataxia. PMID- 12749333 TI - Medical experts and the criminal courts. Clinical forensic medicine needs to become part of the syllabus. PMID- 12749335 TI - [Do politicians feel pain?]. PMID- 12749334 TI - [The activities of Santa Maria Della Scala through 1318 status and its frescoes]. AB - Siena's Hospital was created in the 10th or 11th century, and still owns a rich documentation of his history, stored at the city's state Archives the general organization of the Hospital in the 14th century is accurately described in the 1318 statue. Moreover, during the 15th century frescoes representing its activities were commissioned by the Hospital. They decorated the Pellegrinaio, the main room and the infirmary. The comparison of these two sources from different periods enables us to ask ourselves why the hospital felt compelled to issue what could appear as redundant illustration of the hospital activities. PMID- 12749336 TI - [Health and illness of children in the foundling hospital of Santo Spirito in Sassia in Rome during the XVIII century]. AB - The article deals with the history of the foundling Hospital of S. Spirito in Sassia in Rome during XVIII century. The Hospital sheltered homeless, illegitimate and orphan children, whose health conditions there were often dramatically poor. PMID- 12749337 TI - [The birth of modern hospital]. AB - The modern hospital was born in France, at the end of the XVIII century. The article analyzes the debate promoted by the Academie des Sciences to ascertain what was necessary to rebuild the the hotel Dieu, destroyed by fire. This was the occasion to definitively condemn the medical conception of hospital. PMID- 12749339 TI - Multinational Association of Supportive Cancer Care (MASCC). PMID- 12749338 TI - [The Rome Policlinico Umberto I: an innovative project]. AB - The Rome Policlinico Umberto I is an example of the tendency, already in place at the end of the XIX century, to reunite in a single architectural compound hospital-clinics, labortories, teaching rooms, and libraries, so as to satisfy the needs of University research and education. Guido Baccelli was the promoter of the project: his political and scientific commitment are proved by the imposing architectural structure and by the economic effort that were needed to build the Policlinico. PMID- 12749340 TI - Whole grains health claims in the USA and other efforts to increase whole-grain consumption. AB - In response to the 1990 Nutrition Labeling and Education Act, the Food and Drug Administration approved seven health claims that addressed the relationship between broad food categories and risk of certain chronic diseases. These claims are based on scientific consensus that includes epidemiological, animal and clinical research. The Food and Drug Administration also established a process to petition for new health claims that address substance-disease relationships supported by adequate scientific and specific regulatory requirements. The whole grain-cancer and heart disease authoritative statement health claim approved in July 1999 followed a completely different process mandated by the Food and Drug Administration Modernization Act of 1997. It is based on an authoritative statement made by a government body that represents scientific consensus and is supported by other scientific agencies and organizations. The scientific basis for the claim published in Diet and Health reflects a comprehensive and deliberative review of epidemiological, animal and human studies by the National Academy of Sciences Committee on Diet and Health. Health claims used on whole grain products can attract the attention of health-conscious consumers and are important tools in communicating health messages. However, the US public consumes substantially fewer whole-grain servings than recommended by US dietary guidance. Reasons given by consumers for not purchasing wholegrain foods include colour, price, softness, texture, moisture content and taste. Developing tastier value added wholegrain foods along with simple coordinated messages from industry, the scientific community, public health experts and government will help consumers identify, purchase and consume more wholegrain products. PMID- 12749341 TI - Wholegrain health claims in Europe. AB - Wholegrain foods are important sources of nutrients and phyto-protective components, which are in short supply in many member states of the EU, including the UK. Encouraging the public to increase consumption of wholegrain foods is a positive health message that has critical public health implications. In February 2002 the UK Joint Health Claims Initiative (JHCI) published its authoritative endorsement that wholegrain foods are associated with a healthy heart (Joint Health Claims Initiative, 2002). This new health claim reflects a similar one in the USA based on the accumulation of epidemiological evidence between 1996 and 2001 from several very large cohort studies in the USA, Finland and Norway, which show a consistent protective effect of whole grain and reduced risk of CHD. The JHCI code of practice on health claims requires that the claimed benefit must be scientifically valid, with evidence supporting efficacy of the food in human consumers, under typical conditions of use. The evidence-based approach consists of the identification of studies, an evaluation of individual references, a critical evaluation of the totality of the evidence and a statement that there is significant scientific agreement to establish the validity of the claim. The studies suggest that an intake of three servings per d may have an important cardio-protective effect. The development of a process for the substantiation of health claims in the UK and in the EU is important to underpin regulatory developments, which should protect the consumer, promote fair trade and encourage innovation in the food industry. The present paper sets out the format of the scientific dossier that was presented to the JHCI and includes a call to promote further research to identify the important protective components in the whole grain 'package' and the biological mechanisms behind the observed beneficial effects on health. The major sources of whole grain in the UK are bread and breakfast cereals, and > 90 % of adults in the UK consume less than three servings per d. Increasing the variety and availability of acceptable wholegrain foods could lead to greater consumption levels, which has important public health implications and offers an attractive and food-based dietary strategy for targeting the whole population. PMID- 12749342 TI - Resistant starch as a prebiotic and synbiotic: state of the art. AB - Non-infectious diseases such as CHD and certain cancers have become major causes of death and disability in affluent countries. Probiotics (principally lactic acid bacteria; LAB) may assist in lowering the risk of these diseases. Experimental studies with probiotics have given generally inconclusive outcomes for infectious disease and for biomarkers for non-infectious disease. In part this situation may reflect their inability to colonise the adult human gut effectively. Prebiotics can assist in promoting colonisation, and resistant starch (RS), as a high-amylose starch, is a prebiotic and synbiotic. This starch exerts its synbiotic action through adhesion of the bacteria to the granule surface. Consumption of RS assists in recovery from infectious diarrhoea in man and animals. A rice porridge, high in RS, appears to modify the autochthonous porcine large-bowel microflora favourably through lowering Escherichia coli and coliform numbers. Many of the beneficial effects of RS on large-bowel function appear to be exerted through short-chain fatty acids (SCFA)formed by bacterial fermentation. In man LAB are found in relatively highest numbers in milk-fed infants where the profile of fermentation products differs quite markedly from that in adults. It appears unlikely that ingestion of current probiotics will alter either total SCFA or the proportions of the major acids. More emphasis needs to be given to the investigation of the effects of complex carbohydrates, including RS, on the autochthonous microflora of the human large bowel. PMID- 12749343 TI - Oligosaccharides: state of the art. AB - Oligosaccharides, consisting of a mixture of hexose oligomers with a variable extent of polymerisation, are food products with interesting nutritional properties. They may be naturally present in food, mostly in fruits, vegetables or grains, or produced by biosynthesis from natural sugars or polysaccharides and added to food products because of their nutritional properties or organoleptic characteristics. The dietary intake of oligosaccharides is difficult to estimate, but it may reach 3-13 g/d per person (for fructo-oligosaccharides), depending on the population. The extent of resistance to enzymic reactions occurring in the upper part of the gastrointestinal tract allows oligosaccharides to become 'colonic nutrients', as some intestinal bacterial species express specific hydrolases and are able to convert oligosaccharides into short-chain fatty acids (acetate, lactate, propionate, butyrate) and/or gases by fermentation. Oligosaccharides that selectively promote some interesting bacterial species (e.g. lactobacilli, bifidobacteria), and thus equilibrate intestinal microflora, are now termed prebiotics. The pattern of short-chain fatty acid production in the caeco-colon, as well as the prebiotic effect, if demonstrated, are dynamic processes that vary with the type of oligosaccharide (e.g. extent of polymerisation, nature of hexose moieties), the duration of the treatment, the initial composition of flora or the diet in which they are incorporated. Experimental data obtained in vitro and in vivo in animals, and also recent data obtained in human subjects, support the involvement of dietary oligosaccharides in physiological processes in the different intestinal cell types (e.g. mucins production, cell division, immune cells function, ionic transport) and also outside the gastrointestinal tract (e.g. hormone production, lipid and carbohydrates metabolism). The present paper gives an overview of the future development of oligosaccharides, newly recognised as dietary fibre. PMID- 12749344 TI - Dietary fibre and prebiotics in infant formulas. AB - Prebiotics have the potential to promote immediate and long-term effects on the health and well-being of infants. They have been added to infant formulas in Japan for 20 years and have only recently been used in Europe. The objective is to change the intestinal microflora in order to mimic the bacteriological effect and, thus, the functional effects of human milk. There is, however, a potential risk of long-term effects of the use of these infant formulas. The consequences of using prebiotics in infants, during the first months of life, on the composition and development of the intestinal microflora, and on the resulting bacterial-bacterial and bacterial-host interactions are not known. Attempts have been made to improve infant and follow-on formulas by prebiotic supplementation, but intervention studies are needed to evaluate their immediate and long-term beneficial effects and demonstrate the absence of harmful consequences from their use. PMID- 12749345 TI - Importance of processing for physico-chemical and physiological properties of dietary fibre. AB - There is considerable loss of DM during wet heat treatment of vegetables, leading to an increase in dietary fibre. Correction for the loss of DM indicates that the effects on total dietary fibre are minor. There is, however, depolymerization of the dietary fibre polysaccharides. The degradation is related to the severity of the heat treatment. Souring, freezing and mild microwave treatment have no effects. The viscosity is in general related to the extent of polymerisation. Microwave treatment has different effects on various cultivars of green beans, and the addition of salt (NaCl and CaCl2) to the boiling water changes the physico-chemical properties of soluble fibre in carrots, depending on the cation. The higher viscosity of the soluble fibre in raw carrots may partly explain the lower glucose and hormonal responses observed in healthy subjects when compared with blanched and microwave-cooked carrots. In studies on rats the amount of butyric acid in the distal colon has been shown to be higher with dietary components containing high amounts of resistant starch. Further, the fermentability is lower and the butyric acid concentration higher with composite foods than with the corresponding purified fibre fractions. In human studies the faecal concentration of butyric acid has been shown to increase in patients with ulcerative colitis when [beta-glucan-enriched oat bran (20 g fibre) is added to the diet for 12 weeks. Also, an improvement of symptoms was reported. PMID- 12749347 TI - The glycaemic index: importance of dietary fibre and other food properties. AB - An increasing body of evidence suggests that a low-glycaemic-index (GI) diet has a therapeutic as well as a preventive potential in relation to the insulin resistance syndrome. The implementation of a low-GI diet, however, will require an extended list of low-GI foods to be available on the market. The tailoring of low-GI bread products offers a particular challenge due to their generally high GI and abundance in the diet. Low-GI bread products can be tailored by, for example,enclosure of cereal kernels, sour dough fermentation and/or addition of organic acids, or use of cereal genotypes with elevated contents of amylose or f glucans. Low-GI cereal foods appear to vary in effect on 'second-meal' glucose tolerance in healthy subjects. In addition to the slow-release properties of such foods, the content of dietary fibre appears to play a role. The low glycaemia to starch in a pasta breakfast (GI 54) promoted a higher glucose tolerance and lowered triacylglycerol levels at a standardized lunch ingested 4 h later, compared with a white-wheat-bread breakfast (GI 100). The metabolic benefits of the low GI properties per se have been demonstrated also in the longer term. Thus, a reduction in dietary GI improved glucose and lipid metabolism and normalized fibrinolytic activity in type 2 diabetics, while maintaining a similar amount and composition of dietary fibre. However, the higher dietary fibre content frequently associated with low-GI foods may add to the metabolic merits of a low-GI diet. Consequently, a low-GI barley meal rich in dietary fibre (GI 53) improved glucose tolerance from evening meal to breakfast, whereas an evening meal with pasta had no effect (GI 54). The exchange of common high-GI bread for low-GI high-fibre bread, as the only dietary modification, improved insulin economy in women at risk of type 2 diabetes. These results are in accordance with epidemiological evidence of a reduced risk of type 2 diabetes with a low-GI diet rich in cereal fibre. It is concluded that low-GI cereal foods developed should preferably be rich in dietary fibre. PMID- 12749346 TI - Rye, lignans and human health. AB - Rye bran contains a high content not only of dietary fibre, but also of plant lignans and other bioactive compounds in the so-called dietary fibre complex. Blood concentrations of lignans such as enterolactone have been used as biomarkers of intake of lignan-rich plant food. At present,evidence from studies in human subjects does not warrant the conclusion that rye, whole grains orphyto oestrogens protect against cancer. Some studies, however, have pointed in that direction,especially in relation to cancers of the upper digestive tract. A number of prospective epidemiological studies have clearly shown a protective effect of wholegrain cereals against myocardial infarctions. A corresponding protective effect against diabetes and ischaemic stroke(brain infarct) has also been demonstrated. It seems reasonable to assume that these protective effects are associated with one or more factors in the dietary fibre complex. PMID- 12749348 TI - The active fraction of psyllium seed husk. AB - A series of experiments and evaluations of fractions isolated from psyllium seed husk (PSH) were used to test the overall hypothesis that a gel-forming component of PSH is not fermented and that it is this component that is responsible for the laxative and cholesterol-lowering properties of PSH. A gel is isolated from human stools collected during a controlled diet study when PSH is consumed but not when the control diet only is consumed. Evaluations of three fractions isolated from PSH suggest that gel-forming fraction B, which is about 55% of PSH, is poorly fermented and is the component that increases stool moisture and faecal bile acid excretion, the latter leading to lower blood cholesterol levels. Fraction C, representing < 15% of PSH, is viscous, but is rapidly fermented. Fraction A is alkali-insoluble material that is not fermented. In concentrations comparable with their presence in PSH, fractions A and C do not alter moisture and bile acid output. The active fraction of PSH is a highly-branched arabinoxylan consisting of a xylose backbone and arabinose- and xylose-containing side chains. In contrast to arabinoxylans in cereal grains that are extensively fermented, PSH possesses a structural feature, as yet unidentified, that hinders its fermentation by typical colonic microflora. PMID- 12749349 TI - Extricating handicapped drivers. PMID- 12749351 TI - Astrocytes act as signal transducer in neuronal activation and cerebral arteriole vasodilation. PMID- 12749350 TI - Egas Moniz (1874-1955). PMID- 12749352 TI - ["Tissue engineering": Cultivated heart valves are still a future vision]. PMID- 12749353 TI - [Historical transformation of social attitudes and changements in therapy and assistance of Santa Maria della Pieta Hospital]. AB - In view of a renovation of psychiatric assistance, the authors briefly deal with the history of the social attitude to this issue. Some evidence is taken from the archives of Santa Maria della Pieta, the major institution in Rome for the control of "madness". PMID- 12749355 TI - Children and cancer. PMID- 12749354 TI - [The historical archive of the Psychiatric Hospital of Santa Maria Della Pieta di Roma]. AB - The paper illustrates the archival work on the repository of historical documentation kept in the Hospital (documents from the end of the XIX century to 1978, the year of the Basaglia reform of psychiatric asylums). The paper also deals with the history of the Hospital, 1870 to our days, highlighting the 1913 opening of the 'Nuovo Manicomio Provinciale', and its subsequent evolution and administration, first by the Provincia di Roma then by USL and ASL. PMID- 12749357 TI - The restoration of trust. PMID- 12749356 TI - [Research, training and community links in today's hospitals]. AB - The role of hospitals in research, medical training and their relationship with the community is challenged by socio-economical, epidemiological, demographic and technological changes western health care systems are facing today. This paper enhances that: 1. beside the experimental, high tech and biotechnological model of research, there is still a great need for studies in areas such as: doctor patient relationship, disability, access to care and patient satisfaction; 2. medical training should open to community settings in order to avoid excess in medical specialisation and future shortage of primary health care professionals; 3. a close link with community health care should be established through strategic and operational planning between hospital and community services and a formalised role for family physicians within hospital-based Departments of Family and Community Medicine. PMID- 12749358 TI - Society profile. Interview by Vicki Glaser. PMID- 12749360 TI - Medical Laboratory of the Year. Best little lab by a dam site. PMID- 12749359 TI - Regulation of cellular differentiation and apoptosis by fatty acids and their metabolites. AB - We have reviewed the literature regarding the effects of fatty acids and their metabolites on cellular differentiation and apoptosis. Results obtained in different studies have been variable, but some generalizations can be made. Differentiation was increased by incubation of cells with arachidonic acid (AA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), prostaglandin E1 (PGE1), prostaglandin E2 (PGE2), or leukotriene D4 (LTD4). Effects of these agents on differentiation could be magnified with the simultaneous addition of other differentiation-inducing agents like dimethylsulfoxide or retinoic acid. AA and gamma-linolenic acid increased apoptosis while the effects of n-3 fatty acids (EPA and DHA) and of eicosanoids varied from stimulation to inhibition. These inconsistencies are attributed to the differences in methods used to evaluate differentiation and apoptosis, concentrations of fatty acids and serum, exposure time and the cell models used. Studies using the physiological concentrations of the fatty acids and standardized experimental conditions need to be conducted to establish effects of fatty acids and their metabolites on these cellular processes. PMID- 12749362 TI - Current awareness on yeast. PMID- 12749361 TI - Refrigerators and chillers. PMID- 12749363 TI - Contemplating total artificial heart inactivation in cases of futility. AB - Though currently an experimental technology, there is the potential for implantation of 100,000 total artificial replacement hearts each year in the United States once regulatory approvals are obtained. Although these devices are intended to lengthen life and improve its quality, clinical scenarios can emerge in which the device is no longer serving these goals and termination of life support, including inactivation of the implant, must be contemplated. Although the literature is replete with guidance on the withdrawal of non-implantable therapies, such as dialysis and artificial ventilation, there has been minimal discussion involving the deactivation of implanted therapies. Here, guidance is offered regarding the withdrawal of total artificial heart therapy. PMID- 12749365 TI - More alcohol tax news. PMID- 12749364 TI - Mianserin and trazodone significantly attenuate the intensity of opioid withdrawal symptoms in mice. AB - The aims of this study were to evaluate the effects of trazodone and mianserin on opioid-withdrawal symptoms in morphine-dependent mice. We used a comparative study of the effect of each drug on withdrawal symptoms in one model of acutely high-dose morphine-dependent mice, and two models (high-dose and lu-dose) of chronically morphine-dependent mice at the Tel Aviv University Sackler School of Medicine's laboratory.Trazodone, mianserin or both were given to the morphine dependent mice together with a high dose of naloxone. Intensity of withdrawal symptoms was evaluated by tail-flick assay latencies and three behavioural measurements (rearing, jumping and grooming) in each group. Trazodone and mianserin, each separately,significantly attenuated withdrawal symptoms in all three models. However, the combined treatment of trazodone together with mianserin was not superior to each drug alone. The combination of trazodone and mianserin has no additive value to each drug alone in the control of withdrawal symptoms in opiate-dependent mice undergoing detoxification. When used in clinical settings, caution is needed in order to prevent the unknown influence of opioid-like drugs in medication-assisted detoxification programmes if complete opiate detoxification is the aim. PMID- 12749366 TI - [Current status of tailor-made medicine and the role of clinical pharmacology]. PMID- 12749367 TI - A place apart. PMID- 12749368 TI - The big send-off. PMID- 12749369 TI - Techs-book stuff. PMID- 12749370 TI - Commission possible. PMID- 12749371 TI - Clinical expertise in the era of evidence-based medicine and patient choice. PMID- 12749372 TI - Current awareness in human psychopharmacology. PMID- 12749373 TI - 12th conference on Applied Surface Analysis--AOFA 12. PMID- 12749374 TI - [For a history of medical professions: nursing in modern hospitals]. AB - The article offers an overview about the rapid changes occurring in nursing in recent times. The developments of the nursing profession in the XX century involve bioethical, technical and social issues. PMID- 12749375 TI - Inactivating a total artificial heart: special moral problems. AB - It seems generally correct that patients (or their valid surrogates) should be able to withdraw consent for the use of the total artificial heart (TAH) just as they presently may withdraw consent for other life-prolonging technologies such as the ventilator, but lingering moral problems remain with such decisions. First, should patients be permitted to demand actual removal of the TAH rather than mere deactivation? Second, foregoing other life-prolonging technologies is normally considered "indirect" killing and is therefore judged legal (as well as moral to those who accept only indirect killing). As long as the society includes irreversible stoppage of the heart as one of the criteria for death, however, stopping a TAH will be considered direct killing, i.e., murder. To circumvent this inconsistency, society must change its conclusion that stopping other life prolonging technologies is merely indirect killing, explicitly legalize direct killing by means of stopping a TAH, or revisit the definition of death to eliminate the cardiac standard for death from the definition of death. Assuming that the stopping of the TAH with patient or surrogate consent is acceptable, we must then face the question of whether physicians who believe the TAH is serving no purpose can unilaterally stop the device against the wishes of a patient or surrogate who believes it is still serving a worthwhile purpose. Clinicians should be presumed authoritative in determining the predicted effect of the TAH, but, if the TAH can temporarily prolong life for a patient in a way that is desired by the patient or surrogate, case law, professional society recommendation, and moral analysis all support the conclusion that the TAH must be continued even against the conscientious objection of the physician PMID- 12749376 TI - Replying to Veatch's concerns: special moral problems with total artificial heart inactivation. AB - Moral problems arise when contemplating the inactivation of total artificial heart technology: however, an ethical obligation to explant the device as part of therapy withdrawal is not one of them. Further, arguments will be presented justifying that inactivation of the device is not morally equivalent to active killing of the patient. When device inactivation is clinically and ethically warranted, this decision should not be unilaterally made by the physician but through thorough discussion with the patient (if possible), the patient's surrogate, and the medical team. Consultation with legal counsel and the hospital ethics committee may also be appropriate PMID- 12749377 TI - Many helping hearts: an evaluation of peer gatekeeper training in suicide risk assessment. AB - Literature reviews on suicide prevention programs have presented conflicting results on the efficacy of school-based prevention programs. Gatekeeper training and peer helping are both recommended as part of a comprehensive school-based prevention program, yet there is no literary evidence of the systematic evaluation of gatekeeper training for peer helpers. This study evaluated the efficacy of such training with high school peer helpers using a repeated measures design. Significant gains in knowledge about suicide and skills for responding to suicidal peers were evident immediately after training and 3 months later. There was also a significant improvement in positive attitudes toward suicide intervention following training. Although there was no control group, the research offers tentative support for the efficacy of training peer helpers in suicide risk assessment and indicates the importance of additional training for peer helpers. PMID- 12749378 TI - Death anxiety and attitudes toward the elderly among older adults: the role of gender and ethnicity. AB - The article investigated the relationship between death anxiety, attitudes toward older adults, and personal anxiety toward one's own aging in a group of 197 older men and women. As predicted, negative attitudes toward other older adults were predicted by personal anxieties about aging and death, and, more specifically, fear of the unknown. In addition, several distinctive anxieties were noted for particular subgroups of respondents. Older women scored higher on the Fear of the Dead subscale of the Multidimensional Fear of Death Scale (MFODS) than did men. Caucasian participants displayed higher Fear of the Dying Process than did older African American participants. Lastly, older African American participants reported higher levels of death anxiety on 3 of the subscales of the Multidimensional Fear of Death Scale (Fear of the Unknown, Fear of Conscious Death, and Fear for the Body after Death) when compared with older Caucasian participants and also tended to accord less social value to the elderly. These findings are interpreted in terms of patterns of socialization, and their implications for end-of-life care preferences are noted. PMID- 12749379 TI - Developmental toxicity of ribavirin/IFalpha combination therapy: is the label more dangerous than the drugs? PMID- 12749380 TI - From recognition to responsibility: Josef Warkany, David Smith, and the fetal alcohol syndrome in the 21st century. PMID- 12749381 TI - Developmental toxicology in the 21st century: multidisciplinary approaches using model organisms and genomics. PMID- 12749382 TI - High exogenous homocysteine modifies eye development in early chick embryos. AB - BACKGROUND: Homocysteine is a nonessential aminoacid whose increase is related to the appearance of neural tube defects in humans. In chick embryos, high levels of homocysteine produce neural tube defects and alteration of neural crest cell migration. METHODS: In our study, 8 microl of L-homocysteine thiolatone (20 micromol) was added to chick embryos of Stages 3-8/10 (Hamburger and Hamilton, 1951), (1238 hr of incubation). Three days later, 50 embryos, externally normal or carrying isolated spinal neural tube defects, were sectioned and stained by hematoxilin-eosin or anti-fibrillin-1 antibody. RESULTS: The eye showed alterations of the optic cup as microphthalmia, or lens dislocation. In both cases, the incidence of alterations diminished with the age of the homocysteine increased embryos. Optic cup modifications are probably associated with central nervous system alterations, because most of the affected embryos exhibited isolated spinal neural tube defects and had altered neural crest cells. We have shown for the first time that high exogenous homocysteine during early development could produce a caudally-displaced lens axis before the zonule is formed. Fibrillin-1 is the main component of elastic microfibrils, and in the adult human it is seen as a protein particularly susceptible to homocysteine attack. CONCLUSIONS: Antibody staining against fibrillin-1 showed no evident morphological differences in distribution between experimental and control embryos in the lens, suggesting that fibrillin-1 was not the cause, and malformations may be attributed to other mechanisms. PMID- 12749383 TI - Regulation of AP-2 and apoptosis in developing eye in a vitamin A-deficiency model. AB - BACKGROUND: Eye malformations induced by vitamin A deficiency (VAD) during pregnancy is a major part of the VAD syndrome. But the signaling role of retinoic acid (RA) in ocular tissues is poorly understood. The goal of this study was to determine the role of retinoic acid receptor (RAR) in the development of eye and the possible signaling pathway. METHODS: Time-pregnant mice were treated with 1 mg/kg dose of RAR antagonist AGN193109 (AGN) on 8 days postcoitum (dpc). Newborn mice and 18-dpc embryos were used for phenotype studies. Embryonic eyes of 18 dpc were sectioned for histological study. With immunohistochemistry and TUNEL method, we monitored the alternation of AP-2 expression and apoptotic cells in sections of 12- to 18-dpc embryos. RESULTS: Treatment with AGN resulted in severe craniofacial and eye malformations in virtually all exposed fetuses. The ocular abnormalities included severe defects in anterior segments such as focal corneal thickening and eversion, absence of corneal endothelium and anterior chamber, differentiation defects of lens, as well as defects in posterior segment such as persistent hyperplastic primary vitreous and retinal eversions. The percentage of AP-2-positive cells in ocular tissues on 12, 14, and 18 dpc was significantly (P < 0.05) reduced in AGN-treated eyes compared to control ones. Additionally, the number of apoptotic cell was significantly (P < 0.05) increased in AGN-treated eyes. CONCLUSIONS: The blocking of RAR function can lead to ocular abnormalities that depict partial phenocopies of vitamin A-deficiency syndrome. Both an inhibition of expression of AP-2 and an enhancement of cell death contribute to AGN-induced ocular defects. PMID- 12749384 TI - Expression of cyclooxygenase-2 in embryonic and fetal tissues during organogenesis and late pregnancy. AB - BACKGROUND: Cyclooxygenase (COX) catalyzes the committed step in prostaglandin biosynthesis and exists as two related but unique isoforms, COX-1 (constitutive) and COX-2 (inducible). Prostaglandins (PGs) are known to have many important functions in reproduction, such as placentation and decidualization. Studies with the COX-1 and COX-2 knockout mice have demonstrated that COX-2, but not COX-1, is crucial for normal ovulation, implantation, and decidualization, suggesting that COX-2-derived PGs are important during the initial stages of pregnancy. Although the COX-2 knockout mice did not exhibit any abnormalities at birth, relatively little information exists with regard to the expression of COX-2 in the fetus during development. METHODS: In order to understand the role of COX-2 throughout pregnancy, we characterized the cell type and the temporal expression of inducible COX-2 throughout embryonic and fetal development in the rat (n = 22) by immunohistochemistry and in situ hybridization. RESULTS: High levels of COX-2 expression were seen in decidualized uterine tissue on gestation days 7-13 and then in the fetal membranes on gestation days 17-20. Cyclooxygenase-2 expression was not detectable in any tissues from developing embryos during gestation days 7 13, but was observed in the fetal growth period (gestation days 15-20) in the skin, heart, cartilage, and the kidney. CONCLUSIONS: No COX-2 expression was seen in fetal tissues at days 7-13 of gestation, but was seen in various tissues at days 15-17 of gestation. These observations suggest that COX-2 may be important in mid to late pregnancy through an effect on fetal organ growth, but not in the organogenetic phase of fetal development. PMID- 12749385 TI - Defects in cervical vertebrae in boric acid-exposed rat embryos are associated with anterior shifts of hox gene expression domains. AB - BACKGROUND: Previously, we showed that prenatal exposure to boric acid (BA), an industrial agent with large production, causes alterations of the axial skeleton in rat embryos, reminiscent of homeotic transformations. Indeed, Sprague-Dawley rats exposed in utero to BA on gestation day 9 (GD 9) had only six, rather than the normal seven, cervical vertebrae. This finding, observed in 91% of GD 21 fetuses, suggests posterior transformations of vertebrae. The present study attempts to determine if these skeletal alterations could be explained by modifications of the hox code, involved in the establishment of positional information along the craniocaudal axis of the embryo. METHODS: Pregnant rats were treated by gavage with BA (500 mg/kg, twice) on GD 9. Embryos were collected on GD 11 or GD 13.5 and processed for in situ hybridization. Several hox genes were selected according to the position of their cranial limit of expression in the cervical and thoracic region. RESULTS: At GD 13.5, we detected a cranial shift of the anterior limit of expression of hoxc6 and hoxa6. We observed no difference between control and treated embryos in the location of the cranial limit of expression of the other genes: hoxd4, hoxa4, hoxc5, and hoxa5. CONCLUSIONS: Our results demonstrate that following in utero exposure to BA on GD 9, a disturbance of the expression of hox genes involved inthe specification of most anterior vertebrae is observed at GD 13.5. Based on their expression domain and on their implication in the definition of the cervicothoracic vertebral boundary, it is likely that the anteriorization of hoxc6 and hoxa6 reported here is correlated to the morphological phenotype observed in BA-exposed fetuses at GD 21. PMID- 12749386 TI - Epidemiologic analysis of maternal factors and amniotic band defects. AB - BACKGROUND: The group of defects identified as amniotic bands includes amnion rupture sequence (ARS) and body wall complex (BWC). Little is known about risk factors for either ARS or BWC, except that maternal age has been shown to affect risk inversely. METHODS: The present analysis used data collected from 1976 to 1998 as part of an ongoing case control study of birth defects in the metropolitan areas of Boston, Philadelphia, and Toronto. There were 73 cases with ARS and 11 cases with BWC. ARS cases were further subdivided according to affected structures: there were 53 with only limbs affected (ARS-L) and 20 with nonlimb defects with or without limb defects (ARS-NL). The control group comprised 12,227 subjects with other major malformations. Mothers were interviewed within 6 months of delivery about demographic, reproductive, medical, and behavioral factors. RESULTS: Multivariate adjusted odds ratios for BWC were increased more than threefold for maternal age < 25 years and maternal education < 12 years, but neither estimate was statistically significant. Corresponding estimates for ARS-L and ARS-NL ranged from 13 to 1.5 and also were not statistically significant. Cases were less likely to be white non-Hispanic than controls and the odds ratio for ARS-NL excluded the null. The multivariate adjusted odds ratio (MVOR) for unplanned pregnancy and BWC was 1.9 (95% confidence interval, 0.5-6.7) compared to 1.2 and 1.0 for ARS-L and ARS-NL, respectively. Neither parity nor maternal smoking was associated with any case group. The MVORs for first trimester acetaminophen use in relation to ARS-L and ARS-NL risks were 2.1 (1.1-3.9) and 3.4 (1.1-10.3), respectively. Such use was less common among BWC cases (MVOR was 0.4; 0.1-1.4). CONCLUSIONS: Risk estimates tended to be similar for ARS-L and ARS-NL cases but different for BWC cases, suggesting different etiologies. These data suggest that young maternal age, low maternal education, unplanned pregnancy, and non-white/non-Hispanic race/ethnicity might increase the risk of BWC in offspring. Inceased risks for acetaminophen use should be interpreted with caution because they may be confounded by indication for use. PMID- 12749387 TI - Association between prepregnancy maternal body mass index and the risk of having an infant with a congenital diaphragmatic hernia. AB - BACKGROUND: A previous study observed that women who are underweight prior to conception were significantly more likely to have infants affected by congenital diaphragmatic hernia (CDH). The objective of the current study was to examine the association between maternal body mass index and the risk of a CDH-affected offspring based on a larger number of cases of CDH. METHODS: A case control study was conducted using data collected by the Boston University Slone Epidemiology Center Birth Defects Study, which identifies infants with major malformations who are born in the metropolitan area surrounding Boston, Massachusetts; Philadelphia, Pennsylvania; and Toronto, Ontario, Canada. Control infants were selected from infants without malformations who were born in the same areas. The study included 85 cases of CDH and 655 controls delivered between 1993 and 1997. RESULTS: After adjustment for maternal education and maternal age, we observed that women who were thin or underweight for their height (body mass index < or = 19.0 kg/m2) had an increased risk of having an infant with isolated CDH (OR = 1.9; 95% CI = 0.92-4.1), but no increase in the risk of having a CDH infant with multiple defects (OR = 0.86; 95% CI = 0.29-2.6) compared with all other women. CONCLUSIONS: Although our finding for isolated CDH was not statistically significant (P = 0.08), it suggests that the offspring of thin or underweight women may have an increased risk of isolated CDH. This finding requires confirmation. If it is confirmed, further research on factors associated with being underweight should be explored such as diet, exercise, and use of drugs. PMID- 12749388 TI - Male periconceptional ribavir-ininterferon alpha-2b exposure with no adverse fetal effects. PMID- 12749390 TI - Quantitative assessment of mild-to-moderate coronary atherosclerosis by computerized border detection in 3D IVUS. PMID- 12749389 TI - Validation of an automated system for luminal and medial-adventitial border detection in three-dimensional intravascular ultrasound. AB - The precise tomographic assessment of coronary artery disease by intravascular ultrasound (IVUS) is useful in quantitative studies. Such studies require identification of luminal and medial-adventitial (MA) borders in a sequence of IVUS images. We have developed a three-dimensional (3D) active-surface system for border detection that facilitates the analysis of many images with minimal user interaction. To assess the validity of the technique, luminal and MA borders in 529 end-diastolic images from nine coronary arterial segments (58.8 +/- 14.2 images per patient) were traced manually by four experienced observers. The computer-detected borders were compared with borders determined by the four observers using a modified Williams' index (WI), the ratio of inter-observer variability to computer-observer variability. While manual tracing required 49.2 +/- 12.1 min for analysis, the analysis system identified luminal (R2 = 0.92) and MA borders (R2 = 0.97) in 13.8 +/- 4.0 min, a decrease of 35.4 min (p < 0.000001). The computer minus observer differences in lumen area and MA area were -0.88 +/- 0.90 and -0.07 +/- 0.63 mm2. Therefore, the computer system underestimated both lumen and MA area, but this effect was very small in MA area. The WI values and 95% confidence intervals were 0.98 (0.89,1.06) for luminal border detection and 0.99 (0.95,1.04) for MA border detection. Plaque volume measurements, a common endpoint of clinical trials, also verified the accuracy of the technique (R2 = 0.98). The proposed 3D active-surface border detection system provides a faster and less-tedious alternative to manual tracing for assessment of coronary artery anatomy in vivo. PMID- 12749391 TI - Detection of acute myocardial ischemia during pharmacological stress in graded coronary artery stenosis: analysis of left ventricular asynchrony using Fourier phase imaging in pigs. AB - BACKGROUND: Stress echocardiography is increasingly used to identify coronary artery disease, but quantitative techniques are required to improve the accuracy of this method. Current algorithms used to analyze wall motion usually neglect motion asynchrony that is found in acute ischemia. Fourier phase imaging of echocardiographic images may offer the possibility to detect asynchrony, but its feasibility with dobutamine stress echocardiography (DSE) is undefined. The aim of this study was to investigate whether the extent of left ventricular asynergy can be used to quantify the severity of regional myocardial dysfunction and to detect functionally significant coronary artery stenoses during DSE. METHODS: Regional wall motion abnormalities were induced by graded coronary stenoses (mild and severe) of the left anterior descending coronary artery (LAD) in seven open chest anesthetized pigs. DSE (10-40 microg/kg/min) was performed under control conditions and during sustained ischemia. Coronary flow was measured under resting conditions and during maximal hyperemic response due to intravenous infusion of adenosine. Functional significance of stenoses was defined as mild when coronary flow reserve (CFR) was reduced but > 1.5 and severe when CFR was < or = 1.5. Echocardiographic cine loops were mathematically transformed using a first-harmonic Fourier algorithm displaying the sequence of wall motion as phase angles in parametric images and regional phase histograms. The phase difference (PD) of the first Fourier harmonic of posterior vs. anterior myocardial wall motion was calculated as an index of left ventricular asynchrony. Segmental fractional area shortening (FAS) and wall thickening (WT) as ratio of stress to rest served as a reference method of regional wall motion. RESULTS: The increase in FAS (1.62 +/- 0.6 vs. 0.42 +/- 0.2, p = 0.0002) and WT (1.92 +/- 0.5 to 0.3 +/ 1.1; p = 0.004) in anterior regions during DSE was significantly higher in the control group compared to severe ischemia but not compared to mild ischemia. During graded ischemia, profiles of phase angles were consistently modified, showing a delayed onset in regional contraction. The mean PD during DSE in the control group was 10.4 +/- 7 degrees. PD rose in mild ischemic segments (CFR > 1.5) to 28.9 +/- 10 degrees (p = 0.003) and to 89.6 +/- 25 degrees (p = 0.0002) in severely ischemic segments (CFR < or = 1.5). There was a significant inverse correlation between the PDs and WT (r = -0.87, p < 0.0001). Normal WT ratios yielded low phase angles while segmental phase angles increased with decreased WT. The intraobserver variability of phase analysis was 2.7 +/- 24 degrees (mean +/- 2SD). CONCLUSIONS: These results suggest that left ventricular asynchrony is an indicator of acute ischemia. Echocardiographic Fourier phase imaging is feasible to quantify wall motion displaying contraction sequence in a simple and objective format and is a promising approach for the clinical interpretation of stress echocardiograms. PMID- 12749392 TI - Fourier phase and amplitude analysis for automated objective evaluation of myocardial contrast echocardiograms. AB - AIMS: Objective methods for evaluating myocardial contrast echocardiography (MCE) are not yet widely available. We applied a Fourier analysis to myocardial contrast echocardiograms to identify myocardial perfusion defects. METHODS: Harmonic power-Doppler contrast echocardiograms were performed in 21 patients undergoing Tl-201-SPECT imaging and in 13 controls. Images were transformed using Fourier analysis to obtain phase of the first harmonic sinusoidal curve displayed as color coded sequence of myocardial intensity changes. Means and standard deviations of regional phase angles were measured. The method was validated in an in vitro model. A contrast filled latex balloon was imaged at different gain settings mimicking defined time-intensity curves. An intraoperative porcine infarction model served to prove feasibility of Fourier transformation to analyze real-time pulse inversion contrast echocardiography. RESULTS: In patients, phase imaging and intensity analysis showed focal areas with marked phase shifts (106 +/- 90 degrees) and heterogeneous distribution of phase angles (SD 66 +/- 17 degrees), correctly identifying 13/14 perfusion defects. The in vitro validation yielded increasing phase angles with increasing beta-values. This method was successfully applied to real-time MCE, identifying all infarction areas during occlusion of the left anterior descending artery. CONCLUSION: Phase analysis can be used to display dynamics of myocardial opacification. PMID- 12749393 TI - Prognostic value of nitrate enhanced Tc99m MIBI SPECT study in detecting viable myocardium in patients with coronary artery disease. AB - BACKGROUND: Assessing the viability in akinetic myocardium is vital for predicting functional recovery after therapeutic management in patients with chronic coronary artery disease (CAD) and depressed left ventricular (LV) function. The present study aimed to evaluate the efficacy of Tc99m MIBI SPECT enhanced with nitroglycerine infusion in detecting myocardial viability, as well as to asses the relationship between the myocardial viability and the subsequent treatment and outcome of patients. METHODS AND RESULTS: Sixty-seven consecutive patients with CAD and LV dysfunction (LV ejection fraction 36.6 +/- 8.4%) underwent Tc99m MIBI imaging--at rest and during intravenous nitroglycerine infusion--for viability assessment. Fourteen patients were treated pharmacologically (Group I), and fifty-three (Group II) were submitted to coronary revascularization (PTCR or CABG). Fifteen major cardiac events were observed during 25 months of the follow-up. A significantly worse event-free survival was registered in the subjects of Group I than in Group II subjects. The prognostic predictors of cardiac events were: (1) the number of viable, non revascularized segments in perfusion imaging (p < 0.001), (2) the severity of the disease assessed by coronary angiography (p < 0.05). CONCLUSIONS: Viability detection in nitroglycerine infusion enhanced Tc99m MIBI imaging offers significant prognostic value in patients with CAD after myocardial infarction. Patients with preserved viability showed better prognosis after revascularization than those treated pharmacologically. PMID- 12749394 TI - Evaluation of myocardial viability in chronic ischemic cardiomyopathy. PMID- 12749396 TI - Gadolinium susceptibility artifact causing false positive stenosis isolated to the proximal common carotid artery in 3D dynamic contrast medium enhanced MR angiography of the thorax--a brief review of causes and prevention. AB - Due to the close proximity of arteries and veins in the superior mediastinum and upper extremities, T2* shortening effects of the gadolinium within the central veins may cause artifactual vascular stenosis (susceptibility artifact) on arterial-phase MR angiographic images of the major branches of the aortic arch. We report a case of artifactual stenosis isolated to the origin of the left common carotid artery on arterial-phase MR angiography, secondary to susceptibility artifact from non-diluted gadolinium in the adjacent brachiocephalic vein. The cause of the artifact, its identification and prevention is reviewed. PMID- 12749395 TI - Magnetic resonance imaging of the main pulmonary artery: reliable assessment of dimensions in Marfan patients on a simple axial spin echo image. AB - OBJECTIVE: To investigate if a simple axial spin echo (SE) image can be used for reliable assessment of pulmonary artery dimensions in patients with Marfan syndrome. METHODS: Fifty Marfan patients (mean age 33 +/- 10 years; 34 men, 16 women) and 15 normal subjects (mean age 28 +/- 4 years; nine men, six women) underwent cardiac magnetic resonance imaging (MRI). Pulmonary artery dimensions were obtained on axial SE images at two different levels: (1) the level of the pulmonary artery root, and (2) the level of the pulmonary artery bifurcation. To evaluate the accuracy of axial plane measurements 10 Marfan patients also underwent contrast-enhanced MR angiography (CE-MRA). RESULTS: In the 10 Marfan patients who also underwent CE-MRA, the mean diameter at the pulmonary bifurcation assessed with CE-MRA (31.5 +/- 3.6 mm) was almost equal to mean diameter assessed with axial SE (30.7 +/- 3.6 mm). Agreement of methodology according to Bland and Altman analysis showed a 95% confidence interval ranging from -2.6 to + 4.4 mm for all distances of the pulmonary artery root. In Marfan patients the mean right-left diameter measured on both axial SE images and CE-MRA was approximately 2.5 mm larger than the anterior-right and anterior-left diameters (p < 0.001). CONCLUSIONS: Axial SE MRI is a reliable and easy acquisition to measure pulmonary artery dimensions in patients with Marfan syndrome, and could be used for follow-up, especially in patients with severe involvement of the cardiovascular system. Not only the pulmonary artery trunk but also the asymmetric pulmonary root should be measured, although the clinical relevance of the asymmetric root is not yet known. PMID- 12749397 TI - Prospective navigator gating with a dual acceptance window technique to reduce respiratory motion artifacts in 3D MR coronary angiography. AB - A prospective navigator algorithm with a dual acceptance window (DAW) technique was developed to reduce image artifacts induced by respiratory motion without increasing imaging time. A phantom study shows that the ghost level measured by ghost-to-image ratio was reduced by 27.1 % (p < 0.005) using the DAW technique compared to the conventional single acceptance window technique. This DAW technique can also be used to reduce imaging time while maintaining comparable ghosting level. PMID- 12749398 TI - Detection of repolarization abnormalities in patients with cardiomyopathy using current vector mapping technique on magnetocardiogram. AB - OBJECTIVES: We have previously developed current vector maps of tangential components on the magnetocardiogram (MCG) to obtain cardiac current distribution images. The present study was conducted to detect repolarization abnormalities in patients with cardiomyopathy using the current vector map. SUBJECTS AND METHODS: Thirteen patients with cardiomyopathy (nine males and four females aged 7-16 years, mean, 11.5 +/- 3.1 years, +/- SD), and 15 age- and sex-matched normal subjects were studied. Normal components (Bz) of MCG were measured at rest with a multi-channel superconducting quantum interface device (SQUID) system, and differentiated in the tangential direction to obtain current vector maps. Homogeneity of current in the heart during repolarization was investigated. The direction of the maximum current vector was also calculated in each case. RESULTS: In all normal subjects, the current vector consistently showed a left downward direction on the frontal chest plane during the repolarization process. On the other hand, 8 out of 13 patients with cardiomyopathy showed different patterns; four of these patients showed multi-dipoles, and the other four showed a shift in the current vector direction. One of the eight cases showed no abnormality on electrocardiogram (ECG). CONCLUSIONS: Repolarization process in patients with cardiomyopathy was apparently different from those in normal subjects on the current vector map. It was easy to visualize the repolarization process as a projection to the frontal plane, including regional abnormalities, by the current vector maps, which might be more useful for early detection of repolarization abnormalities than ECG. PMID- 12749399 TI - Guidance of aortic ablation using optical coherence tomography. AB - PURPOSE: There is a significant need for an imaging modality that is capable of providing guidance for intravascular procedures, as current technologies suffer from significant limitations. In particular, laser ablation of in-stent restenosis, revascularization of chronic total occlusions, and pulmonary vein ablation could benefit from guidance. Optical coherence tomography (OCT), a recently introduced technology, is similar to ultrasound except that it measures the back-reflection of infrared light instead of sound. This study examines the ability of OCT to guide vascular laser ablation. METHODS: Aorta samples underwent laser ablation using an argon laser at varying power outputs and were monitored with OCT collecting images at 4 frames. Samples were compared to the corresponding histopathology. RESULTS: Arterial layers could be differentiated in the images sequences. This allowed correlation of changes in the OCT image with power and duration in addition to histopathology. CONCLUSIONS: OCT provides real time guidance of arterial ablation. At 4 frames, OCT was successfully able to show the microstructural changes in the vessel wall during laser ablation. Since current ablation procedures often injure surrounding tissue, the ability to minimize collateral damage to the adjoining tissue represents a useful advantage of this system. This study suggests a possible role for OCT in the guidance of intravascular procedures. PMID- 12749400 TI - Tumor extending through inferior vena cava into the right atrium. A late recurrence of renal cell carcinoma. AB - Renal cell carcinoma is a tumor with a distinct feature; it can invade through renal vein to the inferior vena cava (IVC) and growths intravascularly extending sometimes into the right cardiac chambers. Previous reports of renal cancer with extension to the right atrium at the time of diagnosis have been published. We present the case of a patient with a right atrial mass arising from the IVC found by echocardiography, which was subsequently proved to be a recurrence of renal cell carcinoma 5 years after radical nephrectomy. PMID- 12749401 TI - Cytotoxic 4'-aminochalcones and related compounds. AB - A series of 4'-aminochalcones 1 and related maleamic acids 2 and Schiff bases 3 were designed and synthesized as candidate cytotoxic agents. The atomic charges on different atoms of representative compounds were calculated. Evaluation of the enones 1-3 against human Molt 4/C8 and CEM T-lymphocytes as well as murine P388 and L1210 leukemic cells revealed that approximately 40% of the IC50 values generated were less than 10 microM. In some cases cytotoxicity was correlated with the Hammett sigma values of the aryl substituents and less frequently with the aryl Hansch pi values. Evidence was obtained that in general these compounds displayed selective toxicity for certain malignant cells and were well tolerated in mice. This study has revealed various directions whereby the project may be amplified in the future with a view to finding compounds with increased cytotoxicity to tumour cells. PMID- 12749402 TI - Synthesis and pharmacological investigation of some novel 2,3-disubstituted quinazolin-4(3H)-ones as analgesic and antiinflammatory agents. AB - A series of novel 2-substituted quinazolin-4(3H)-ones have been synthesized by condensing the aromatic primary amine of 2-substituted quinazolines with different aldehydes and ketones. The synthesized compounds were confirmed by their spectral data (IR, NMR and MS) and the purity was ascertained by elemental analysis. When these compounds were evaluated for analgesic and antiinflammatory activities, compounds I-VIII exhibited more potent analgesic activity than diclofenac sodium, while the compounds IV, V, VI and VII exhibited more potent antiinflammatory activity than diclofenac sodium. The activity values were found to be significant as compared to those of controls. PMID- 12749403 TI - Synthesis, pharmacological activity and chromatographic separation of some novel potential beta-blockers of the aryloxyaminopropanol type. AB - Following our previous structure-activity relationship studies, some novel compounds of the aryloxyaminopropanol type, derived from 2- or 4 hydroxyphenylalkanones, with phenethyl or 3,4-dimethoxyphenethyl groups in the hydrophilic part of the molecule were synthesized and pharmacologically evaluated. The compounds were prepared by means of two methods and their structures were confirmed by the interpretation of their IR, UV and 1H NMR spectra. The enantiomers were separated by HPLC on vancomycin (Chirobiotic V) and teicoplanin (Chirobiotic T) chiral stationary phases. The affinity of the prepared racemic compounds to beta1- and beta2-adrenergic receptors was pre determined on isolated guinea pig atria and trachea. The assumed cardioselectivity was expressed as the beta1/beta2 ratio. Reciprocal changes in the position of the phenoxysubstituents did not influence the antiisoprenaline activity of the compounds. On the other hand, the increase of the N-substituent size in the hydrophilic part of molecule (3,4-dimethoxyphenethyl moietyled to a substantially higher affinity for cardiac (beta1) than for tracheal (beta2) tissue. PMID- 12749405 TI - Study and analytical application of ion-pair formation in the system fluoxetine pyrocatechol violet and fluvoxamine-pyrocatechol violet. AB - Pyrocatechol violet (PCV) reacts in aqueous media with fluoxetine (FLX) and fluvoxamine (FLV) forming coloured ion-association complexes, which are insoluble in water but quantitatively extracted into chloroform-n-butanol mixture. The composition of the compounds, studied by spectrophotometric methods showed that the molar ratio PCV : FLX and PCV : FLV is 1 : 1. The compounds were characterized by UV-VIS, IR and NMR spectrometry. Under optimal experimental conditions fluoxetine and fluvoxamine were determined in the range 1.3-18.0 microg/ml and 2.6-39.1 microg/ml, respectively. The proposed methods have been succesfully applied to the determination of these drugs in pharmaceuticals and natural samples. PMID- 12749404 TI - HPLC assay in plasma and preliminary pharmacokinetic study of a quinazolinone derivative with spasmolytic effect. AB - An analytical method for analysis of 3-(o-bromo)-phenyl-2-(2',3' dihydroxypropylthio)-4(3H)-quinazolinone from rat plasma using HPLC with reversed phase C18 and liquid-liquid extraction was developed. This method was used for a pharmacokinetic study in rat. PMID- 12749406 TI - Kinetics of degradation of quinapril hydrochloride in tablets. AB - The effect of temperature and of humidity on the stability of quinapril hydrochloride (QHCI) in tablets as well as on the durability of the degradation process was investigated. The investigation has been performed by means of the "enhanced ageing" test (in the temperature range from 383 to 333 K, at a relative humidity between and 76%). From the results of the study it follows that QHCI in tablets undergoes decomposition according to a first order reaction model. The activation energies as well as enthalpy and entropy (for temperature 298 K) have also been determined. It is concluded that the decomposition of QHCI in tablets proceeds by way of hydrolysis of the ester group and an intramolecular cyclization. PMID- 12749408 TI - Development and optimization of the drug release determination of iron from iron prolonged release tablets. AB - The aim of this research was to develop and optimize the drug release determination of iron from iron prolonged release tablets. First, solubility characteristics of ferrous sulfate in different aqueous media with a pH in the range of 1 to 8 were studied. According to the results obtained different physicochemical conditions that influence drug release of iron from iron prolonged release tablets were checked. Various aqueous media with a pH in the range of 1 to 7.4, different volumes of dissolution media, various rotation speeds of stirring elements, different dissolution apparatus (apparatus 1/basket apparatus and apparatus 2/paddle apparatus according to USP/Ph. Eur.) were verified. For evaluation of dissolved iron two different methods were checked: atomic absorption spectrometry and redox titration. Redox titration was proved to be a reproducible, discriminatory, selective and cost effective method for evaluation of dissolved iron from iron prolonged release tablets and can be applied in the quality control of solid dosage forms containing iron compounds. PMID- 12749407 TI - Proteolytic enzyme conjugated to SC-glucan as an enzymatic transdermal drug penetration enhancer. AB - The objective of this study was to investigate the effect of papain, a proteolytic enzyme, on the percutaneous absorption of drugs. To guarantee the enzyme stability during the skin penetration, papain was modified by the conjugation to SC-glucan. The enhancing activity of drug penetration was evaluated using antipyrine and indomethacin as hydrophilic and hydrophobic model drugs, respectively. The SC-glucan-papain conjugate was found to be very effective for facilitating the percutaneous absorption of antipyrine. Microscopic observations showed that the thickness of stratum corneum and viable epidermis was increased by the treatment of the SC-glucan-papain conjugate. Moreover, it induced phase separation, lacuna formation, and lamellar disruption within the stratum corneum interstices. These structural changes by the SC-glucan-papain conjugate are likely to be induced from hydrolysis of extensive crosslinking of corneocyte envelopes and intracellular proteins. However, the SC-glucan-papain conjugate showed no skin irritation according to the Draize test, which may be due to the difficulty of the SC-glucan-papain conjugate in penetrating into the skin. PMID- 12749409 TI - Studies on gynaecological hydrophilic lactic acid preparations. Part 5: The use of Eudragit E-100 as lactic acid carrier in intravaginal tablets. AB - Intravaginal tablets based on hydrophilic methylcellulose and containing lactic acid complexed with Eudragit E-100 undergo deformation under standard conditions. The high flow--limit of gel originating from the tablets as well as its dynamic viscosity should enable durability of this dosage form on the vaginal mucosa. By selecting either 1 : 1 or 2 : 1 ratios of lactic acid to Eudragit E-100 it is possible to obtain tablets that disintegrate into a gelform at pH 3.8-4.4, i.e. the pH remains within the physiological range. Increasing the amount of lactic acid in the complex in relation to the polymer to a 4:1 ratio results in gels with a lower pH while possessing an acid reserve that can neutralize the excess of alkali present in severe vaginal infections. PMID- 12749411 TI - Inhibitory effect of Bergenia ligulata on influenza virus A. AB - Methanol water extract from rhizomes of Bergenia ligulata, a plant used in Nepalese ethnomedicine, inhibited in vitro the replication of influenza virus in a dose dependent manner and did not show virucidal activity at effective concentration. Pretreatment of cells with B. ligulata extract was shown to be most effective to prevent cell destruction. The extract inhibited viral RNA synthesis and reduced viral peptide synthesis at 10 microg/ml. The virus inhibitory effect is related to the presence of condensed tannins in the extract. PMID- 12749412 TI - 12alpha-hydroxystigmast-4-en-3-one: a new bioactive steroid from Toona ciliata (Meliaceae). AB - In a bioassay guided phytochemical investigation of Toona ciliata (Fam. Meliaceae), a new hydroxy steroidal ketone, 12alpha-hydroxystigmast-4-en-3-one (1) was isolated from the petroleum ether extract of the plant together with two known steroids and three C-methyl coumarins. The structure of 1 was established by means of spectroscopic analyses. Compound 1 was found cytotoxic in a brine shrimp lethality bioassay with LC50 of 9.9 microg/ml and it also showed significant antitumor activity with Ti50 value of 14.1 microg/ml in a potato disc bioassay. PMID- 12749410 TI - Evaluation of the mutagenicity of antimalarial products isolated from Solanum nudum (Solanaceae). AB - Diosgenone is a major component of the hexane extract from the plant Solanum nudum (Solanaceae). The products from degraded and acetylated diosgenone that showed in vitro antimalarial activity against the FCB-2 strain of Plasmodium falciparum and methanol, dichloromethane and ethereal extracts of Solanum nudum were tested for their mutagenic activity using the Ames test with the TA-97a, TA 98, TA-100 and TA-102 strains of Salmonella typhimurium. These compounds were not mutagenic at the tested concentrations. PMID- 12749413 TI - Anatomical study on Vaccinium arctostaphylos L. AB - Vaccinium arctostaphylos L. (Ericaceae), locally named Qaraqat, is widely used in Iranian folk medicine as antidiabetic and antihypertensive agents. The anatomical study on the different organs of the plant was performed to give a clear standard for identification of the drug. Various diagnostic elements such as particular cells, rubiaceous stomata, covering and glandular trichomes, Ca-oxalate crystals were found in different parts. PMID- 12749415 TI - Effects of polysorbates on antiviral and antibacterial activity of monoglyceride in pharmaceutical formulations. PMID- 12749414 TI - Validated HPLC method for the determination of ranitidine in human serum and its application in a clinical pharmacokinetic study. PMID- 12749416 TI - The critical micellar concentration of derivatives of piperidinoethylesters of 2 alkoxyphenylcarbamic acid. Study of local anesthetics. Part 162. PMID- 12749417 TI - Inhibitors of farnesyltransferase: 5-arylacryloylaminobenzophenones show antimalarial activity. PMID- 12749418 TI - Role of reactive oxygen species in the pathophysiology of human reproduction. AB - OBJECTIVE: To summarize the role of reactive oxygen species (ROS) in the pathophysiology of human reproduction. DESIGN: Review of literature. SETTING: Fertility research center and obstetrics and gynecology department in a tertiary care facility. RESULT(S): ROS plays an essential role in the pathogenesis of many reproductive processes. In male-factor infertility. oxidative stress attacks the fluidity of the sperm plasma membrane and the integrity of DNA in the sperm nucleus. Reactive oxygen species induced DNA damage may accelerate the process of germ cell apoptosis, leading to the decline in sperm counts associated with male infertility. ROS mediated female fertility disorders share many pathogenic similarities with the ones on the male side. These similarities include a potential role in the pathophysiology of endometriosis and unexplained infertility. High follicular fluid ROS levels are associated with negative IVF outcomes, particularly in smokers. Moreover, oxidative stress may be responsible in hydrosalpingeal fluid mediated embryotoxicity as well as poor in vitro embryonic development. CONCLUSION(S): High levels of ROS are detrimental to the fertility potential both in natural and assisted conception states. PMID- 12749420 TI - Infertility medications and the risk of breast cancer. PMID- 12749419 TI - Infertility drugs and the risk of breast cancer: findings from the National Institute of Child Health and Human Development Women's Contraceptive and Reproductive Experiences Study. AB - OBJECTIVE: To determine the association between infertility drug use and invasive breast cancer in a population-based case-control study. DESIGN: Multicenter case control study. SETTING: Women aged 35 to 64 years in metropolitan Atlanta, Detroit, Los Angeles, Philadelphia, and Seattle. PATIENT(S): The 4,575 case patients had histologically confirmed primary invasive breast cancer. The 4,682 control subjects were women without breast cancer identified in the same geographic locations using randomized-digit dialing. INTERVENTION(S): A standardized questionnaire focusing on reproductive health and family history as well as use of oral contraceptives and other hormones and infertility drugs was administered to all subjects. Data on the type of breast cancer were also obtained. MAIN OUTCOME MEASURE(S): Odds ratios examining the association between use of various infertility drugs and invasive breast cancer. RESULT(S): Overall, a history of infertility drug use was not associated with the risk of developing breast cancer. Compared with women who never used any fertility medication, however, women using human menopausal gonadotropin (hMG) for > or = 6 months or for at least six cycles had a relative risk of breast cancer ranging between 2.7 to 3.8. CONCLUSION(S): Long-term use of certain infertility drugs could adversely affect risk of breast cancer. Additional confirmatory studies are needed. PMID- 12749421 TI - Hormonal contraception can suppress natural antimicrobial gene transcription in human endometrium. AB - OBJECTIVE: To determine the effect of hormonal contraception with a combined oral contraceptive pill and levonorgestrel intrauterine system on the expression of the natural antimicrobials secretory leukocyte protease inhibitor, beta-defensins 1 and 2, and granulysin in human endometrium. DESIGN: Observational study. SETTING: Day case ward in a department of obstetrics and gynecology. PATIENT(S): Fifty seven women undergoing gynecologic procedures for benign conditions; 24 received no contraception for more than 3 months, 20 received a combined oral contraceptive for more than 3 months, and 13 wore a levonorgestrel intrauterine system for more than 3 months. MAIN OUTCOME MEASURE(S): Endometrial samples were collected from all women. Messenger RNA was extracted and quantitative polymerase chain reaction was used to investigate expression of secretory leukocyte protease inhibitor, beta-defensin 1, beta-defensin 2, and granulysin. Immunohistochemistry for secretory leukocyte protease inhibitor was performed. RESULT(S): All antimicrobials varied cyclically. The level of secretory leukocyte protease inhibitor was maximal in the late secretory and menstrual phase, beta-defensin 1 in the mid secretory phase, granulysin in the late secretory phase, and beta defensin 2 in the menstrual phase. Use of a combined oral contraceptive or levonorgestrel intrauterine system use decreased messenger RNA expression of beta defensin 1 and 2 and granulysin but not secretory leukocyte protease inhibitor. CONCLUSION(S): Endogenous and exogenous sex-steroid hormones, in the form of a combined oral contraceptive or levonorgestrel intrauterine system, influence gene transcription of secretory leukocyte protease inhibitor, beta-defensin 1, beta defensin 2, and granulysin in the endometrium. PMID- 12749422 TI - Contraceptive activity of a spermicidal aryl phosphate derivative of bromo methoxy-zidovudine (compound WHI-07) in rabbits. AB - OBJECTIVE: To determine the vaginal contraceptive activity of WHI-07 in the rabbit model. DESIGN: Prospective, controlled study. SETTING: Center for advanced preclinical sciences. ANIMAL(S): Subgroups of 15, 16, or 24 New Zealand White does and 24 bucks per experiment. INTERVENTION(S): Ex vivo (Experiment 1) and in vivo (Experiments 2 and 3) treatment of semen with WHI-07 or Nonoxynol-9 (N-9). In Experiment I, ovulated does in subgroups of 15 were artificially inseminated with semen mixed with WHI-07 or vehicle. In Experiment 2, ovulated does in subgroups of 24 were artificially inseminated within 2 min after intravaginal administration of 2% WHI-07 gel-microemulsion or 2% N-9 gel and allowed to complete term pregnancy. In Experiment 3, ovulated does in subgroups of 16 were artificially inseminated at 15, 30, or 60 minutes. MAIN OUTCOME MEASURE(S): The numbers of implanted embryos on postinsemination day 8 or the proportion of does that became pregnant and delivered newborn rabbits; the litter size, weight, growth, and viability of pups until lactation day 5. RESULT(S): Exposure of semen to WHI-07 at the time of artificial insemination completely inhibited pregnancy rates (WHI-07-pretreated, 0%, vs. control, 60%) and embryo implantation (WHI-07 pretreated, 0/175 vs. control, 68/170). Intravaginal administration of a 2% WHI 07 gel-microemulsion or 2% N-9 gel before artificial insemination significantly inhibited pregnancy rates (81% and 85% inhibition, respectively) when compared with control. Furthermore, the 2% WHI-07 gel-microemulsion provided >90% inhibition of fertility even when insemination was delayed until 60 minutes after intravaginal application. Rabbits that delivered litters despite intravaginal application of 2% WHI-07 gel-microemulsion had healthy offsprings with no perinatal or postnatal repercussions. CONCLUSION(S): WHI-07 is a potent contraceptive agent in vivo. Intravaginal use of WHI-07 gel-microemulsion has clinical potential as a safe prophylactic contraceptive, in addition to its microbicide activity to curb the sexual transmission of HIV. PMID- 12749423 TI - Exposure to high levels of luteinizing hormone and estradiol in the early follicular phase of gonadotropin-releasing hormone antagonist cycles is associated with a reduced chance of pregnancy. AB - OBJECTIVE: To compare ongoing implantation rates under two different GnRH antagonist protocols. DESIGN: Randomized controlled trial. SETTING: Tertiary referral center. PATIENT(S): One hundred eleven women undergoing ovarian stimulation for in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI). INTERVENTION(S): Ovarian stimulation with 150 IU recombinant-FSH (rec FSH) starting on day 2 of the cycle and GnRH antagonist starting either on day 6 of stimulation (fixed group) or when a follicle of > or = 15 mm was present after at least 5 days of stimulation (flexible group). In the flexible group, the rec FSH dose was increased to 250 IU when the antagonist was initiated. MAIN OUTCOME MEASURE(S): Ongoing implantation and pregnancy rate. RESULT(S): In patients with no follicle of > or = 15 mm present on day 6 of stimulation, a significantly lower ongoing implantation rate was observed if the flexible scheme was applied as compared with the fixed scheme of administration (8.8% vs. 23.9%, respectively). Exposure of the genital tract to LH or E2 from initiation of stimulation to antagonist administration was able to distinguish between pregnant and nonpregnant patients in the population studied. CONCLUSION(S): High exposure of the genital tract to LH and E2 in the early follicular phase is associated with a reduced chance of pregnancy in cycles stimulated with recombinant FSH and GnRH antagonist for IVF/ICSI. PMID- 12749424 TI - Subcutaneous versus intramuscular administration of human chorionic gonadotropin during an in vitro fertilization cycle. AB - OBJECTIVE: To confirm that hCG levels in follicular fluid and serum would be comparable between i.m. and s.c. administration of purified hCG. DESIGN: In a prospective study, serum and follicular fluid levels of hCG after an i.m. or s.c. injection of 10,000 IU of hCG were evaluated 36 hours after injection, that is, at the time of oocyte retrieval. SETTING: This study was carried out in a university-affiliated IVF program. PATIENT(S): Forty women undergoing oocyte retrieval were entered into the study at the time of egg retrieval, that is, 36 hours after hCG administration. INTERVENTION(S): S.c. or i.m. injection of hCG. MAIN OUTCOME MEASURE(S): Serum and follicular fluid concentrations of hCG were evaluated 36 hours after injection at the time of oocyte retrieval. RESULT(S): There was a significantly higher serum hCG level in the s.c. group (348.6 +/- 98 IU/L) vs. the i.m. group (259.0 +/- 115 IU/L) and a significantly higher follicular fluid hCG level in the s.c. vs. the i.m. group (233.5 +/- 85 vs. 143.4 +/- 134 IU/L). CONCLUSION(S): After purified hCG administration via the s.c. route, both serum and follicular fluid levels are greater compared with the i.m. route. PMID- 12749425 TI - Prospective evaluation of two stimulation protocols for low responders who were undergoing in vitro fertilization-embryo transfer. AB - OBJECTIVE: To compare two stimulation protocols designed for low responders undergoing IVF. DESIGN: Randomized, prospective study. SETTING: University hospital IVF unit. PATIENT(S): Sixty low responders who were recruited on the basis of results in previous cycles. INTERVENTION(S): Modified flare protocol in which a high dose of GnRH agonist was administered for the first 4 days, followed by a standard agonist dose, or a modified long protocol in which a standard agonist dose was used until pituitary down-regulation, after which the agonist dose was halved during stimulation. MAIN OUTCOME MEASURE(S): Number of oocytes retrieved. RESULT(S): Twenty-nine cycles were performed with the modified flare protocol and 31 were performed with the modified long protocol. Significantly more oocytes were obtained with the modified long protocol than the modified flare protocol (4.42 +/- 2.6 vs. 3.07 +/- 2.15). The number and quality of embryos available for transfer was similar in both groups. One clinical pregnancy (3.4%) was achieved with the modified flare protocol, and 7 pregnancies (22.5%) were achieved using the modified long protocol. CONCLUSION(S): These preliminary results substantiate the poor prognosis and outcome for low responders undergoing IVF. A modified long "mini-dose" protocol appears to be superior to a modified mega-dose flare protocol in terms of oocyte yield and cycle outcome. PMID- 12749426 TI - Transfer of frozen-thawed blastocysts that underwent quarter laser-assisted hatching at the day 3 cleaving stage before freezing. AB - OBJECTIVE: To examine the pregnancy potential of frozen-thawed blastocysts that underwent quarter laser-assisted hatching (AH) at the cleaving stage before freezing and to compare clinical and embryo characteristics between the groups that succeeded in and failed to achieve pregnancy. DESIGN: Prospective observational study. PATIENT(S): Thirty-four of 112 patients with frozen blastocysts requiring transfer of thawed embryos between January 2000 and December 2001. SETTING: Assisted reproductive technology unit in a tertiary medical center. INTERVENTION(S): Embryos of patients undergoing blastocyst transfers routinely underwent quarter AH using a nontouch 1.48-microm diode laser. Blastocysts not transferred were cryopreserved using a six-step freezing protocol with glycerol as the cryoprotectant. MAIN OUTCOME MEASURE(S): Postthaw embryo survival and zona pellucida (ZP) maintenance, implantation rate, and clinical pregnancy rate per transfer of thawed blastocysts. RESULT(S): A total of 118 frozen blastocysts was thawed. Of these, 89 (75.4%) embryos survived and were transferred in 35 cycles. Assisted hatching-manipulated ZP tolerated the freeze thaw procedures without shape distortion in surviving embryos. Eleven (31.4%) clinical pregnancies with 15 intrauterine gestational sacs occurred, for an implantation rate of 16.9%. Those who succeeded in pregnancy tended to have more embryos available before freezing, more original blastocysts of good quality for freezing, and more surviving blastocysts after thawing for transfer than did those who failed to achieve pregnancy. CONCLUSION(S): An acceptable clinical pregnancy rate was obtained from transfer of thawed blastocysts that underwent laser AH on the ZP at the day 3 cleaving embryo stage in fresh cycles in selected patients. Embryo characteristics before freezing played major roles in determining implantation potential of thawed embryos. PMID- 12749427 TI - Assessment of leukemia inhibitory factor levels by uterine flushing at the time of egg retrieval does not adversely affect pregnancy rates with in vitro fertilization. AB - OBJECTIVE: To assess intrauterine levels of leukemia inhibitory factor (LIF) by uterine flushing at the time of egg retrieval and to confirm that the procedure has no detrimental effect on pregnancy rates. DESIGN: Prospective study. SETTING: Assisted reproductive unit of a university hospital. PATIENT(S): Uterine flushing was performed in 148 IVF patients. The first 100 patients were compared with a matched control group. INTERVENTION(S): Uterine flushing at the time of egg retrieval. MAIN OUTCOME MEASURE(S): IVP-ET results, pregnancy rates, and intrauterine LIF levels. RESULT(S): Pregnancy rates were not different in the group of patients with (27%) or without uterine flushing (28%). Leukemia inhibitory factor was detected in 60 patients (46%). Pregnancy rates did not differ between patients' detectable LIF and those in whom LIF was undetectable. Mean levels of LIF were 30.1 +/- 49.3 pg/mL and 28.6 +/- 51.2 pg/mL in pregnant and nonpregnant patients respectively. CONCLUSION(S): The flushing procedure at the time of egg retrieval did not adversely affect pregnancy rates. Leukemia inhibitory factor was detected in 46% of patients at the time of egg retrieval, but no correlation were observed with better pregnancy rates in patients with detectable LIF. Mean LIF levels did not differ in pregnant and nonpregnant women. Access to endoluminal secretions of the endometrium during IVF-ET may represent a new research in human implantation. PMID- 12749428 TI - Effects of low concentrations of inhibin B on the outcomes of testicular sperm extraction and intracytoplasmic sperm injection. AB - OBJECTIVE: To study the effects of undetectable inhibin B concentrations on the outcomes of testicular sperm extraction (TESE) and of intracytoplasmic sperm injection (ICSI). DESIGN: Retrospective study. SETTING: Obstetrics, gynecology, and reproductive biology departments. PATIENT(S): We carried out TESE on 75 men with nonobstructive azoospermia: 42 men had an inhibin B concentration of or = 15 pg/mL (group 2). Twenty-five ICSI cycles were carried out using sperm from men in group 1 (group A1), and 35 ISCI cycles were carried out using sperm from men in group 2 (group A2). The outcomes of ICSI in groups A1 and A2 were compared with those of 81 ICSI cycles performed for obstructive azoospermia (group B). INTERVENTION(S): Testicular sperm extraction, testicular spermatozoa cryopreservation, and ICSI. MAIN OUTCOME MEASURE(S): Testicular sperm extraction outcome, pregnancy, and delivery. RESULT(S): Sperm were significantly less likely to be successfully recovered from men in group 1 than from those in group 2 (21% vs. 48%). The inhibin B concentration was significantly lower in men in whom TESE failed, but the FSH concentration did not differ. The implantation rate per embryo transferred was twofold lower in group A1 (7.4%) than in group B (16%), but this difference is not statistically significant. CONCLUSION(S): Patients with undetectable inhibin B concentration should be informed of the low chances of positive testicular biopsy, and more embryos should be transferred to improve the success rate. PMID- 12749429 TI - Sexual dysfunction in men undergoing infertility evaluation: a cohort observational study. AB - OBJECTIVE: To study psychosexual problems in men undergoing infertility evaluation. DESIGN: A cohort observational study. SETTING: Male infertility diagnostic center. PATIENT(S): Four hundred twelve men undergoing infertility evaluation between 1999 and 2001. INTERVENTION(S): Baseline and follow-up data on sexual functions. Semen analysis for samples collected by masturbation. A second analysis was requested in 2 weeks upon finding an abnormality of semen parameters. Penile vibratory stimulation to help men who failed to collect semen on their second or subsequent appointments. MAIN OUTCOME MEASURE(S): Sexual functions (erection and orgasm) during the time of infertility evaluation. RESULT(S): Seven of 412 men were excluded from the analysis due to a past history of sexual dysfunction. Of the remaining 405 men, 46 (11%) failed to collect semen by masturbation for a second semen analysis after repeated (2-4 times) attempts at 2- to 3-day intervals. Nine of these men (20%) were able to collect semen using vibratory stimulation. All 46 men experienced problems with erection or orgasm and had severe anxiety during attempts to masturbate and during sexual contact with their partners. CONCLUSION(S): Our study indicates that some men may experience sexual dysfunction of a psychogenic nature in response to the diagnosis of infertility. PMID- 12749430 TI - Meiotic behavior of the sex chromosomes in a 45,X/46,X,r(Y)/46,X,dic r(Y) patient whose semen was assessed by fluorescence in situ hybridization. AB - OBJECTIVE: To determine the meiotic behavior of a ring Y chromosome in a semen sample from a 45,X/46,X,r(Y)/46,X,dic r(Y) patient and the possible interchromosomal effects of the ring on other chromosome pairs. DESIGN: Retrospective analysis. SETTING: Universitat Autonoma de Barcelona. PATIENT: An oligoasthenoteratozoospermic patient who presented for infertility consultation. MAIN OUTCOME MEASURE(S): The sex chromosome content of spermatogenic cells, meiotic figures, and spermatozoa in the ejaculate and the possible interchromosomal effects on chromosomes 13, 18, and 21 were analyzed by using multicolor fluorescence in situ hybridization. Germ-cell aneuploidies were scored. RESULT(S): X0 cells are meiotically incompetent. All meiotic figures were exclusively XY, and 80% showed unpaired sex chromosomes. A high proportion of postreductional cells were XY (45.5%) or nullisomic for sex chromosomes (13.92%). This percentage decreased in spermatozoa to 14.89% and 27.66%, respectively. A statistically significant increase in X-bearing versus Y-bearing cells both in postreductional cells (23.9% vs. 14.3%) and spermatozoa (41.9% vs. 19.3%) was also observed. Evidence for an interchromosomal effect on chromosome 21 was detected. CONCLUSION(S): Data suggest that this patient had a generalized increase incidence of chromosome anomalies, underscoring the importance of incorporating screening for sperm aneuploidies in genetic analysis of affected patients. PMID- 12749431 TI - Relationship between serum leptin concentrations and bone mineral density as well as biochemical markers of bone turnover in women with postmenopausal osteoporosis. AB - OBJECTIVE: To determine whether leptin is involved in bone remodeling in patients with postmenopausal osteoporosis. DESIGN: Cross-sectional study. SETTING: Department of Obstetrics and Gynecology, Faculty of Medicine, Cairo University. PATIENT(S): Ninety postmenopausal osteoporotic women (37 obese and 53 nonobese) and 30 healthy premenopausal women from the same clinic served as controls. Lumbar spine bone mineral density (LS-BMD) of osteoporotic patients was more than 2.5 SD below the normal mean of healthy premenopausal women. MAIN OUTCOME MEASURE(S): Serum levels of leptin, osteocalcin (OC), bone alkaline phosphatase (B-ALP), urinary deoxypyridinoline (DPyr), and N-telopeptide of type 1 collagen (NTX) as well as LS-BMD using dual energy X-ray absorptiometry (DEXA). RESULT(S): The serum leptin level in obese postmenopausal osteoporotic patients was significantly increased compared with nonobese osteoporotic patients. There were no significant differences of bone formation markers (B-ALP, OC), bone resorption markers (DPyr, NTX), or LS-BMD between the obese and nonobese groups. There were no significant correlations between serum leptin and any biomarkers of bone turnover and BMD. CONCLUSION(S): In postmenopausal osteoporotic patients with increased bone turnover, serum leptin concentration is not correlated with BMD or with the biomarkers of bone formation or bone resorption. PMID- 12749432 TI - Correlation of androgen receptors, aromatase, and 5-alpha reductase in the human vagina with menopausal status. AB - OBJECTIVE: To determine whether aromatase and 5alpha-reductase mRNAs are expressed in human vagina and to evaluate the presence of androgen receptors in human vaginal tissue based on age and menopausal status. DESIGN: Prospective study. SETTING: Specimens obtained from clinical renal urology practice. PATIENT(S): Premenopausal and postmenopausal women undergoing surgery for prolapse or incontinence. MAIN OUTCOME MEASURE(S): Expression of aromatase and 5alpha-reductase type 1 and 2 mRNAs was evidenced by reverse transcriptase polymerase chain reaction (RT-PCR), and the density of androgen receptors was measured by semiquantitative immunohistochemistry. RESULT(S): The mRNAs for aromatase and 5alpha-reductase isotypes 1 and 2 were detected in vaginal specimens. Androgen receptors were present in vaginal mucosa, submucosa, stroma, smooth muscle, and vascular endothelium. Expression was significantly greater in vaginal submucosa. A negative correlation existed between age and androgen receptor density. CONCLUSION(S): Expression of genes encoding for enzymes involved in testosterone metabolism in the human vagina, as well as androgen receptor location, density, and changes with menopausal status, suggests that androgens may play a role in regulating vaginal smooth muscle and vaginal blood flow. PMID- 12749434 TI - Evaluation of the impact of laparoscopic ovarian drilling on Doppler indices of ovarian stromal blood flow, serum vascular endothelial growth factor, and insulin like growth factor-1 in women with polycystic ovary syndrome. AB - OBJECTIVE: To study the serum levels and correlation of vascular endothelial growth factor (VEGF), insulin-like growth factor 1 (IGF-1), hormonal profile, and Doppler blood flow changes within the ovarian stroma before and after laparoscopic ovarian drilling (LOD) in women with clomiphene-resistant polycystic ovary syndrome (PCOS). DESIGN: Prospective controlled study. SETTING: University teaching hospital. PATIENT(S): Twenty-five women with clomiphene-resistant PCOS (group 1) and 20 women with regular menstrual cycles as a comparison group (group 2). INTERVENTION(S): Laparoscopic ovarian drilling. MAIN OUTCOME MEASURE(S): Serum levels of VEGF, IGF-1, and Doppler indices of ovarian stromal blood flow. RESULT(S): The serum levels of VEGF, IGF-1, T, and LH were significantly higher in group 1 before LOD than in group 2. The Doppler indices (pulsatility index and resistance index) of ovarian stromal blood flow were also significantly lower in group 1 before LOD than in group 2. The serum levels of VEGF, T, and LH were significantly reduced in group 1 after LOD compared with in group 1 before LOD. Doppler indices (pulsatility index and resistance index) of ovarian stromal blood flow were significantly increased after LOD. The VEGF levels before LOD were positively correlated with IGF-1, LH, and T. After LOD, the VEGF levels were positively correlated with LH and T. CONCLUSION(S): Higher serum levels of VEGF and IGF-1 may explain the increased vascularity that was demonstrated by Doppler blood flow measurements in PCOS. Laparoscopic ovarian drilling reduced serum VEGF, IGF-1, T, and LH and reduced ovarian blood flow velocities, which may explain the reduction of ovarian hyperstimulation syndrome in women with PCOS after LOD. PMID- 12749433 TI - Troglitazone decreases adrenal androgen levels in women with polycystic ovary syndrome. AB - OBJECTIVE: To determine whether amelioration of insulin resistance in polycystic ovary syndrome (PCOS) with the insulin sensitizer troglitazone (TGZ) decreases circulating adrenal androgens (AAs), as reflected by DHEAS levels. DESIGN: Prospective, randomized, double-blind clinical trial. SETTING: Multicenter study. SUBJECT(S): Three-hundred five women with PCOS. INTERVENTION(S): Subjects were randomly assigned to receive either placebo (PBO; n = 73) or TGZ in doses of 150 mg/day (TGZ-150; n = 78), 300 mg/day (TGZ-300; n = 77), or 600 mg/day (TGZ-600; n = 77) for 20 weeks. Blood was sampled before (week 0) and at week 20 of treatment. MAIN OUTCOME MEASURE(S): DHEAS, insulin, and glucose levels were determined in the blood samples. RESULT(S): There were no differences in age, body mass, or racial composition among the groups. Our results indicate that basal insulin declined in a dose-related fashion. Likewise, TGZ administration caused a dose-related decrease in DHEAS levels. To detect extreme effects, we subsequently subdivided patients receiving PBO or TGZ-600 into tertiles according to initial DHEAS levels. Patients receiving PBO in the lowest (n = 27) and highest (n = 22) DHEAS tertiles experienced a 16.8% +/- 62.0% and a -11.1% +/- 17.4% change in DHEAS levels during the study, respectively. Alternatively, patients with PCOS receiving TGZ-600 in both the lowest (n = 29) and the highest (n = 23) DHEAS tertiles experienced a drop in DHEAS levels (-18.7% +/- 27.2% and 26.4% +/- 17.2%, respectively), a significant difference from PBO. CONCLUSION(S): In conclusion, improving the insulin resistance-related hyperinsulinemia of PCOS with TGZ results in a decrease in DHEAS levels, regardless of initial DHEAS level. Whether the observed suppression is the direct result of decreased insulin levels or whether it reflects other direct and indirect effects of TGZ remains to be determined. PMID- 12749435 TI - Use of cyproterone acetate, finasteride, and spironolactone to treat idiopathic hirsutism. AB - OBJECTIVE: To compare the effectiveness of cyproterone acetate, finasteride, and spironolactone in the treatment of idiopathic hirsutism. DESIGN: Prospective randomized clinical study. SETTING: University hospital. PATIENT(S): Forty-one women (median age, 21 years [range, 18-34 years]) with idiopathic hirsutism who had requested to use an oral contraceptive. INTERVENTION(S): Patients were randomly assigned to receive cyproterone acetate (12.5 mg/d for the first 10 days of the cycle), finasteride (5 mg/d), or spironolactone (100 mg/d) for 12 months. Follow-up was done at the end of therapy. MAIN OUTCOME MEASURE(S): Ferriman Gallwey score before treatment, at 6 and 12 months of treatment, and 1 year after the end of treatment, and androgenic profile before and after treatment. RESULT(S): At the end of therapy, the Ferriman-Gallwey score decreased by 38.9%, 38.6%, and 38.5% in patients who used cyproterone acetate, finasteride, and spironolactone, respectively. One year after therapy, the Ferriman-Gallwey score of patients who used spironolactone was significantly lower (6.74 +/- 1.41) than that of patients who used either cyproterone acetate (7.92 +/- 1.08), or finasteride (9.08 +/- 0.99). The androgenic profile did not change significantly during treatment. CONCLUSION(S): In patients with idiopathic hirsutism, the short term results of treatment with cyproterone acetate, finasteride, and spironolactone are similar, but spironolactone is effective for a longer time. PMID- 12749436 TI - Hyperandrogenicity is an alternative mechanism underlying oligomenorrhea or amenorrhea in female athletes and may improve physical performance. AB - OBJECTIVE: To evaluate endocrine mechanisms underlying oligomenorrhea or amenorrhea in female athletes. DESIGN: Cross-sectional study. SETTING: Women's health clinical research unit at a university hospital. PATIENT(S): Age- and BMI matched groups of athletes active in endurance sports with and without menstrual disturbances and regularly cycling sedentary controls. INTERVENTION(S): Groups were compared with respect to endocrine status, body composition, and physical performance. MAIN OUTCOME MEASURE(S): Identification of a subgroup of oligomenorrheic or amenorrheic athletes with increased androgen levels and anabolic body composition. RESULT(S): A subgroup of 8 of 25 athletes with menstrual disturbances had significantly higher serum levels of free and total testosterone, androstenedione, LH-FSH ratio, and lower SHBG levels than did all other groups. Other oligomenorrheic or amenorrheic athletes had normal values comparable to those in regularly menstruating athletes and controls. The hyperandrogenic subgroup showed a more anabolic body composition, with higher total bone mineral density and upper-lower fat mass ratio than did oligomenorrheic or amenorrheic athletes with normal androgen levels. The hyperandrogenic subgroup had the highest VO2 max and the highest performance values in general. CONCLUSION(S): Menstrual disturbances in female athletes are often explained as a consequence of hypothalamic inhibition and caloric deficiency. We suggest that essential hyperandrogenism is an alternative mechanism underlying oligomenorrhea or amenorrhea in some female athletes and may imply an advantage for physical performance. PMID- 12749437 TI - Metformin has direct effects on human ovarian steroidogenesis. AB - OBJECTIVE: To investigate the possibility of direct effects of metformin on ovarian steroidogenesis. DESIGN: Cultured ovarian cells. SETTING: Academic research environment. PATIENT(S): Women undergoing bilateral salpingoophorectomy for benign gynecological disease. MAIN OUTCOME MEASURE(S): Estradiol and P were measured in granulosa cell (GC) conditioned medium and androstenedione (A) and P in theca conditioned medium. RESULT(S): The effect of addition of metformin alone to GCs was variable, but significant inhibition of both P and E2 was seen (range 0%-30%). Metformin dose-dependently inhibited gonadotrophin and insulin stimulated P and E2 production (range 25%-50%). In theca, metformin inhibited A production (0%-40%) with no effect on P. In the presence of insulin, A was inhibited dose-dependently and P increased by a similar magnitude. CONCLUSION(S): These results demonstrate a direct effect of metformin on ovarian steroidogenesis. The inhibitory effects on androgen production in particular would be beneficial in polycystic ovary syndrome (PCOS). PMID- 12749438 TI - Therapeutic effects of the levonorgestrel-releasing intrauterine system in the treatment of idiopathic menorrhagia. AB - OBJECTIVE: To investigate the effect of the levonorgestrel-releasing intrauterine system (LNG-IUS) in the treatment of idiopathic menorrhagia. DESIGN: Measurements of menstrual blood loss (MBL), hemoglobin, and serum ferritin before and after LNG-IUS insertion. SETTING: National Research Institute for Family Planning and Beijing Gynecology and Obstetrics Hospital, Beijing, People's Republic of China. PATIENT(S): Thirty-four patients with MBL over 80 mL. INTERVENTION(S): Insertion of the LNG-IUS on cycle days 5-7 and follow-up at 3-month intervals for 3 years. MAIN OUTCOME MEASURE(S): Measurement of MBL, serum ferritin, and hemoglobin for evaluation of efficacy of treatment. RESULT(S): A significant reduction of MBL to 23.4 mL (78.7% decrease), 26.4 mL (83.8% decrease), 2.7 mL (97.7% decrease), and 13.7 mL (85.0% decrease) at 6, 12, 24, and 36 months, respectively. After 6 months, one-third of the patients experienced amenorrhea, and one-fourth, spotting. Hemoglobin increased significantly from 121.5 g/L preinsertion to 135.5 g/L after 36 months, while serum ferritin levels increased significantly from 21.9 ng/mL before insertion to 92.8 ng/mL after 36 months. In women using the LNG IUS for 3-4 years, the E2 levels in 20 samples were 239.4 pmol/L, P levels were 11.1 nmol/L, and serum LNG levels were maintained at an average of 511 pmol/L. CONCLUSION(S): The significant reduction of MBL and the increase in hemoglobin and serum ferritin levels in the treatment of menorrhagia with the LNG-IUS has great implications for women's reproductive health, particularly in developing countries. PMID- 12749439 TI - Temporal and morphologic characteristics of pinopod expression across the secretory phase of the endometrial cycle in normally cycling women with proven fertility. AB - OBJECTIVE: To assess the temporal and morphologic characteristics of pinopod expression on the surface of endometrium across the secretory phase, in LH-timed endometrial samples in normal, healthy women. DESIGN: Prospective, randomized study. SETTING: Academic teaching hospital. PATIENT(S): Sixty-eight healthy volunteers with proven fertility. INTERVENTION(S): Urinary LH-timed endometrial and blood sampling was performed on each subject on a randomly selected day of the secretory phase. MAIN OUTCOME MEASURE(S): Histologic dating, assessment of pinopods using scanning electron microscopy, and comparison with serum P levels. RESULT(S): Eighty-six endometrial tissue samples obtained from 68 subjects were evaluated under scanning electron microscopy. Pinopods were first observed on luteal day 5, corresponding with the onset of the midluteal phase increase in serum P levels. Pinopods persisted for the entire duration of the secretory phase, but their morphology changed as the cycle advanced. CONCLUSION(S): The present findings demonstrate that pinopods are a characteristic feature of the mid to late secretory phase endometrial epithelium, exhibit cycle-dependent changes in morphology, and are most prominent during the putative implantation interval. PMID- 12749440 TI - Serum concentrations of interleukin-2R (IL-2R), IL-6, IL-8, and tumor necrosis factor alpha in patients with ectopic pregnancy. AB - OBJECTIVE: To investigate the diagnostic relevance of serum cytokine concentrations in ectopic pregnancy (EP). DESIGN: Cohort study. SETTING: University hospital. PATIENT(S): Seventeen women with EP, 22 women with miscarriage, and 33 women with normal intrauterine pregnancy, at comparable stages of gestation. INTERVENTION(S): Interleukin (IL)-2 receptor, IL-6, IL-8, and tumor necrosis factor alpha (TNF-alpha) determination by immunoradiometric assay. MAIN OUTCOME MEASURE(S): Serum concentrations of progesterone, beta-hCG, IL-2R, IL-6, IL-8, and TNF-alpha. RESULT(S): Serum levels of IL-6 were higher in women with EP than in those with miscarriage and normal pregnancy. Serum levels of TNF-alpha were higher in women with EP than in those with miscarriage and normal pregnancy. Serum levels of IL-8 were higher in women with EP than in those with miscarriage and normal pregnancy. An IL-8 cutoff of >40 pg/mL predicted EP with a sensitivity of 82.4%, a specificity of 81.8%, and positive and negative predictive values of 58.3% and 93.8%. No difference in serum IL-2R levels was found among the groups. CONCLUSION(S): Serum IL-8, IL-6, and TNF-alpha concentrations are higher in women with EP than in those with miscarriage and normal pregnancy. Further studies are needed to determine their diagnostic value. PMID- 12749441 TI - Human chorionic gonadotropin level as a predictor of trophoblastic infiltration into the tubal wall in ectopic pregnancy: a blinded study. AB - OBJECTIVE: To determine the relationship between gestational age, tubal ultrasonographic diameter, and serum hCG levels and different stages of trophoblastic infiltration of the tubal wall in ectopic pregnancy. DESIGN: Blinded prospective study. SETTING: University-based clinic in Italy. PATIENT(S): Thirty-seven consecutive patients with an ampullary ectopic pregnancy. INTERVENTION(S): Laparoscopic salpingectomy. MAIN OUTCOME MEASURE(S): Gestational age, diameter of the tubal mass as determined by transvaginal ultrasonography. and hCG level on the day of surgery. Ectopic pregnancy was classified according to the depth of trophoblastic infiltration: trophoblast limited to the tubal mucosa (stage I), extension to the tubal muscularis (stage II), or complete tubal wall infiltration up to the serosa discontinued by trophoblastic cells (stage III). RESULT(S): Fifteen patients (40.5%) had stage I tubal infiltration, 14 (37.8%) had stage II infiltration, and 8 (21.6%) had stage III infiltration. Gestational age and diameter of the tube did not differ among the three groups. The median hCG level was 1,710.5 mIU/mL (range, 113-5,635 mIU/mL) for patients with stage I infiltration. 4,690.0 mIU/mL (range, 150-21,531 mIU/mL) for patients with stage II infiltration, and 15,700.0 mIU/mL (range, 13,809-21,650 mIU/mL) for patients with stage III infiltration. All the patients with hCG levels > 6,000 mIU/mL had stage II or III invasion. CONCLUSION(S): These findings may explain why the conservative treatment of ectopic pregnancy is less successful in patients with high hCG levels than in patients with low levels. Use of radical procedures may be justified in the former group. PMID- 12749442 TI - Plasminogen activator/plasminogen activator inhibitor-1 and cytokine modulation by the PROACT System. AB - OBJECTIVE: To examine the effects of the PROACT treatment on the fibrinolytic system and inflammatory cytokines in human peritoneum. DESIGN: Controlled clinical study. SETTING: University hospital. PATIENT(S): Nine subjects undergoing laparotomy had peritoneal samples taken at the incision. INTERVENTION(S): The PROACT applicator was inserted through the peritoneal incision, and treatment of peritoneum was performed twice. A peritoneal sample was taken from one treated area. At closure, the second treated sample and an additional control sample were taken. All four samples were snap frozen in liquid nitrogen. Samples were homogenized and protein content extracted. MAIN OUTCOME MEASURE(S): Concentrations of total and active transforming growth factor-beta 1 (TGF-beta1), tumor necrosis factor-alpha (TNF-alpha), tissue-type plasminogen activator (t-PA), urokinase plasminogen activator (uPA), and plasminogen activator inhibitor 1 (PAI-1) were obtained. RESULT(S): Total TGF-beta1 at opening was 30% less in treated samples. At closure, active TGF-beta1 increased significantly (163%) in control samples and not in treated samples. Tumor necrosis factor alpha was detectable only in control samples at closure. During surgery, tPA levels showed a marked decrease in control samples vs. a small increase in treated samples. Levels of uPA increased significantly only in the control samples. In control samples, tPA/PAI-1 ratio was two thirds of treated sample ratio. CONCLUSION(S): Heating of the peritoneum with the PROACT System modulates the biologic tissue response to induce effects that would be consistent with inhibition of postoperative adhesion development. PMID- 12749444 TI - Use of strict sonohysterographic methods for preoperative assessment of submucous myomas. AB - OBJECTIVE: To assess the diagnostic accuracy of sonohysterography (SHG) and transvaginal sonography versus diagnostic hysteroscopy in preoperative assessment of submucous myomas. DESIGN: Prospective pilot study. SETTING: University hospital outpatient center. PATIENT(S): Forty-eight symptomatic (bleeding, infertility) premenopausal patients with submucous myomas. INTERVENTION(S): Preoperative grading of submucous myomas with a strict SHG methodology and standard transvaginal sonography compared with hysteroscopic grading of submucous myoma before hysteroscopic myomectomy. MAIN OUTCOME MEASURE(S): SHG and sonographic agreement with hysteroscopic findings. RESULT(S): Forty-eight patients were enrolled (mean age +/- SD = 41 years +/- 10.2). The median duration of SHG was 12 minutes (interquartile range, 9-16). The mean number of submucous myomas was 1 (range, 1-3) per woman. In all cases, a successful SHG was performed, with no, mild, or moderate pain in 38 (79%), 8 (17%), and 2 (4%) patients, respectively. No patients experienced severe pain or vasovagal reaction. All cases were correctly diagnosed by SHG compared with the final hysteroscopic diagnosis (kappa = 1.0; SE = 0.105). Simple transvaginal ultrasound was inaccurate in six cases (kappa = 0.81; SE = 0.103). CONCLUSION(S): Strict and reproducible SHG diagnostic procedures proved to be as effective as hysteroscopy and well tolerated in preoperative grading of submucous myomas. PMID- 12749443 TI - Use of carbon dioxide versus normal saline for diagnostic hysteroscopy. AB - OBJECTIVE: To compare CO2 and normal saline as distention medium in diagnostic hysteroscopy. DESIGN: Prospective randomized study. SETTING: University-based artificial insemination and sterility center in Italy. PATIENT(S): Seventy-four women who underwent hysteroscopy. INTERVENTION(S): Hysteroscopy was performed with CO2 or normal saline. MAIN OUTCOME MEASURE(S): Quality of intrauterine images, cervical dilatation, local anesthesia, and duration of the test. In addition, each patient evaluated pain during and after the examination, irritation of the phrenic plexus, analgesic use, and side effects. RESULT(S): The quality of the hysteroscopic image was statistically similar for both media. Dilatation of the cervical canal and use of local anesthesia was more often necessary in the CO2 group. The procedure time was 5.96 +/- 1.55 minutes in the CO2 group and 3.12 +/- 0.96 in the normal saline group. The CO2 group reported pain more frequently during and after the examination, shoulder pain, greater analgesic use, and more side effects. CONCLUSION(S): For hysteroscopy, normal saline is technically equal to CO2 in terms of image quality and ease of use but offers more advantages. Hysteroscopy with normal saline is more acceptable to patients, quicker to perform, and entails fewer additional procedures. PMID- 12749445 TI - Association between the length of the vas deferens excised during vasectomy and the risk of postvasectomy recanalization. AB - OBJECTIVE: To evaluate the association between the length of the vas deferens excised during vasectomy and the risk of recanalization. DESIGN: Nested case control study. SETTING: Hospital-based Family Planning Clinic in Quebec City (Canada). PATIENT(S): Among 870 vasectomized men, all 47 cases of spontaneous recanalization and 188 controls whose first semen analysis showed either azoospermia (controls A) or <1 x 10(6)/mL nonmotile sperm (controls B). MAIN OUTCOME MEASURE(S): Spontaneous recanalization defined as a semen analysis showing any motile sperm 6 weeks or more after vasectomy. RESULT(S): Individual vas segments excised ranged from 5 to 20 mm in 227 (97%) of the 235 participants. The mean +/- SD of the average of both segments for each man was 12 +/- 4 mm, identical in cases and in controls. In cases and controls A, the risk ratio (95% confidence interval [CI]) of recanalization with an average of segments of <10 mm and 10-14 mm was 0.6 (0.1-2.0) and 0.6 (0.2-1.6) when compared to 15 mm or more, respectively. In cases vs. controls B, the risk ratio of recanalization was 1.6 (0.4-7.7) and 0.6 (0.2-1.7), respectively. CONCLUSION(S): In this cohort, there was no association between the length of vas segment excised and the risk of recanalization. PMID- 12749446 TI - Success of laparoscopic ovarian wedge resection is related to obesity, lipid profile, and insulin levels. AB - OBJECTIVE: To evaluate the effects of laparoscopic ovarian wedge resection on hormonal and metabolic parameters of polycystic ovary syndrome (PCOS) and to compare profiles of women who achieved pregnancy with those who did not. DESIGN: Prospective study. SETTING: University hospital. PATIENT(S): Thirty-three women with PCOS. INTERVENTION(S): Laparoscopic ovarian wedge resection using harmonic scalpel. MAIN OUTCOME MEASURE(S): Pregnancy; levels of testosterone, DHEAS, gonadotropins, sex hormone-binding globulin (SHBG), and glucose and insulin during 2-hour glucose tolerance test; lipid profile; body mass index; and waist to-hip ratio. RESULT(S): Twenty-two women (67%) achieved clinical pregnancy within the mean of 4.9 months after surgery. Baseline parameters of women who became pregnant differed from those who did not: those who became pregnant were less obese, had lower levels of total cholesterol, low-density lipoprotein, and triglycerides; higher levels of SHBG; lower levels of fasting insulin; lower insulin area under the curve; and higher insulin sensitivity index. Subjects not pregnant by 12 weeks after surgery underwent repeat endocrine and metabolic evaluations. In these women, wedge resection was followed by declines in testosterone, LH, and insulin sensitivity index. Wedge resection had no significant effect on SHBG, DHEAS, or lipid profile. CONCLUSION(S): Laparoscopic wedge resections are associated with the highest pregnancy rates among less obese subjects with favorable lipid profiles and lower insulin. In this study, the postoperative decline of serum testosterone and LH is not attributable to improvement of insulin sensitivity. PMID- 12749447 TI - Ultrasonography as a predictor of embryo implantation after in vitro fertilization: a controlled study. AB - OBJECTIVE: To investigate the predictive value of ultrasonographic parameters as prognostic indicators of implantation after IVF when measured on the day of embryo transfer. DESIGN: Comparative, observational study. SETTING: University teaching hospital. PATIENT(S): Two hundred eighty patients undergoing IVF. INTERVENTION(S): Ovarian stimulation, IVF. MAIN OUTCOME MEASURE(S): Variables related to patients' clinical characteristics, treatment characteristics, ovarian response, ovum retrieval, outcome of IVF and ICSI, embryo transfer, ultrasonographic and Doppler endometrial measurements, and uterine blood flow that have been proposed as potential predictive factors of implantation. All transvaginal ultrasonographic assessments were performed on the day of embryo transfer. RESULT(S): Among 240 patients finally evaluable, 67 (group 1) became pregnant after IVF, and 173 (group 2) failed to conceive. The 111 nonpregnant patients who had the same embryo score per replacement (group 3) as did patients in group 1 were selected for comparison purposes. The only significant differences between groups 1 and 3 were the type A endometrium and the absence of a protodiastolic notch in the uterine arteries, both of which were more frequently found in group 1. However, a considerable overlap existed between conception and nonconception cycles regarding both variables. CONCLUSION(S): Ultrasonographic parameters as predictors of implantation in assisted reproduction have a limited value in the clinical setting. PMID- 12749448 TI - Sonovaginography is a new technique for assessing rectovaginal endometriosis. AB - OBJECTIVE: To evaluate the efficacy of a new technique, the sonovaginography, for the assessment of rectovaginal endometriosis. DESIGN: Prospective study. SETTING: University hospital. PATIENT(S): Forty-six women were scheduled for laparotomic or laparoscopic surgery because of rectovaginal endometriosis suspected on the basis of patient history and/or clinical examination. INTERVENTION(S): Before surgery, all the women underwent transvaginal ultrasonography and then sonovaginography. The latter is based on transvaginal ultrasonography combined with the introduction of saline solution to the vagina that creates an acoustic window between the transvaginal probe and the surrounding structures of the vagina. Ultrasound findings were compared with the results of surgical exploration and histological examination. MAIN OUTCOME MEASURE(S): We assessed the accuracy of transvaginal ultrasonography and of sonovaginography for the detection and the location and extension assessment of rectovaginal endometriotic lesions, as well as compared patient compliance between the procedures. RESULT(S): Sonovaginography diagnosed rectovaginal endometriosis more accurately than did transvaginal ultrasonography, with a sensitivity and specificity of 90.6% and 85.7%, respectively, whereas the transvaginal ultrasonography has shown a sensitivity and specificity of 43.7% and 50%, respectively. Patient discomfort did not differ significantly between the procedures. CONCLUSION(S): Sonovaginography is a reliable and simple method for the assessment of rectovaginal endometriosis and provides information on location, extension, and infiltration of the lesions, which are important factors in selecting the kind of surgery. PMID- 12749449 TI - Novel treatment of a patient with secondary infertility due to retained fetal bone. AB - OBJECTIVE: To describe a simple and previously unreported treatment for retained fetal bone fragments as a cause of secondary infertility. SETTING: Fertility center at a Canadian teaching hospital. DESIGN: Case report. PATIENT(S): A 36 year-old woman with a 15-year history of secondary infertility. INTERVENTION(S): A second dilation and curettage (D+C) performed under abdominal ultrasound guidance was performed where the curette could be directed for the removal of echogenic endometrial foci. MAIN OUTCOME MEASURE(S): Resolution of long-term infertility. RESULT(S): Spontaneous pregnancy 4 months after ultrasound-guided D+C and subsequent term delivery. CONCLUSION(S): If an echogenic area is discovered in the endometrium, it is now standard to look at the uterine cavity by hysteroscopy. However, if the hysteroscopy is normal, we suggest that a D+C with intraoperative abdominal ultrasound assistance be done to ensure that all the abnormal tissue is removed. PMID- 12749450 TI - Significance of one pronucleus before fertilization. AB - OBJECTIVE: To describe a case of primary infertility associated with oocytes having one pronucleus before fertilization on repeated IVF attempts. DESIGN: Case report. SETTING: A university-based assisted reproduction unit. PATIENT(S): A 30 year-old woman with primary infertility and oocytes containing one pronucleus before fertilization. INTERVENTION(S): Oocyte donation. MAIN OUTCOME MEASURE(S): Pregnancy. RESULT(S): Conceived triplets after transfer of three embryos using donor oocytes. CONCLUSION(S): This patient's infertility was likely associated with an oocyte abnormality, as evidenced by the premature formation of one pronucleus before fertilization. In the future, more studies on the appearance of a single pronucleus before fertilization will be needed to determine its overall significance on fertility. PMID- 12749451 TI - Peritonitis after a ruptured left pyosalpinx in a patient undergoing in vitro fertilization. AB - OBJECTIVE: To report acute abdomen in a young woman with infertility due to small asymptomatic bilateral hydrosalpinges who was on the 12th day of an IVF cycle. DESIGN: Case report. SETTING: Private IVF clinic. PATIENT(S): A 35-year-old woman with asymptomatic bilateral hydrosalpinges. INTERVENTION(S): Abdominal laparoscopy. MAIN OUTCOME MEASURE(S): Pus and a ruptured left pyosalpinx. RESULTS: The patient suddenly developed acute abdomen while lifting a disabled person at a nursing home where she worked as a nurse. About 200 mL of pus was aspirated, and the patient had a ruptured left pyosalpinx. Bilateral salpingectomy was done by laparoscopy; the patient's postoperative recovery was uneventful. CONCLUSION: We report a rare case of acute abdomen due to peritonitis after a ruptured pyosalpinx in an IVF patient who lifted a disabled person. PMID- 12749452 TI - Use of recombinant follicle-stimulating hormone (Gonal F) and recombinant luteinizing hormone (Luveris) for multiple follicular stimulation in patients with a suboptimal response to in vitro fertilization. PMID- 12749453 TI - Power enough? Confidence intervals for uncertainty. PMID- 12749454 TI - Power enough? Confidence intervals for uncertainty. PMID- 12749455 TI - Power enough? Confidence intervals for uncertainty. PMID- 12749456 TI - "Floating denominators"--effect of verification bias on accuracy estimates for preimplantation diagnosis. PMID- 12749457 TI - "Floating denominators"--effect of verification bias on accuracy estimates for preimplantation diagnosis. PMID- 12749458 TI - Total reproductive potential of a single cycle--to include fresh and frozen embryos? PMID- 12749459 TI - Risks of monochorionic pregnancies after assisted hatching? PMID- 12749460 TI - Conceptual interrelatedness and caricatures. AB - Concepts are interrelated to the extent that the characterization of each concept is influenced by the other concepts, and are isolated to the extent that the characterization of one concept is independent of other concepts. The relative categorization accuracy of the prototype and caricature of a concept can be used as a measure of concept interrelatedness. The prototype is the central tendency of a concept, whereas a caricature deviates from the concept's central tendency in the direction opposite the central tendency of other acquired concepts. The prototype is predicted to be relatively well categorized when a concept is relatively independent of other concepts, but the caricature is predicted to be relatively well categorized when a concept is highly related to other concepts. Support for these predictions comes from manipulations of the labels given to simultaneously acquired concepts (Experiment 1) and of the order of categories during learning (Experiment 2). PMID- 12749461 TI - On the generality of optimal versus objective classifier feedback effects on decision criterion learning in perceptual categorization. AB - Biased category payoff matrices engender separate reward- and accuracy-maximizing decision criteria Although instructed to maximize reward, observers use suboptimal decision criteria that place greater emphasis on accuracy than is optimal. In this study, objective classifier feedback (the objectively correct response) was compared with optimal classifier feedback (the optimal classifier's response) at two levels of category discriminability when zero or negative costs accompanied incorrect responses for two payoff matrix multiplication factors. Performance was superior for optimal classifier feedback relative to objective classifier feedback for both zero- and negative-cost conditions, especially when category discriminability was low, but the magnitude of the optimal classifier advantage was approximately equal for zero- and negative-cost conditions. The optimal classifier feedback performance advantage did not interact with the payoff matrix multiplication factor. Model-based analyses suggested that the weight placed on accuracy was reduced for optimal classifier feedback relative to objective classifier feedback and for high category discriminability relative to low category discriminability. In addition, the weight placed on accuracy declined with training when feedback was based on the optimal classifier and remained relatively stable when feedback was based on the objective classifier. These results suggest that feedback based on the optimal classifier leads to superior decision criterion learning across a wide range of experimental conditions. PMID- 12749462 TI - Domain differences in the structure of artifactual and natural categories. AB - In three experiments, different methodologies, measures, and items were employed to address the question of whether, and to what extent, membership in a semantic category is all or none (i.e., absolute) or a matter of degree (i.e., graded). Resemblance theory claims that categorization is based on similarity, and because similarity is graded, category membership may also be graded. Psychological essentialism asserts that categorization is based on the presumption of the category essence. Because artifactual (e.g., FURNITURE) and natural (e.g., FRUIT) categories have different sorts of essences, artifacts and natural kinds may be categorized in qualitatively different manners. The results converged onthe finding of a robust domain difference in category structure: Artifactual categories were more graded than natural categories. Furthermore, typicality reliably predicted absolute category membership, but failed to predict graded category membership. These results suggest that resemblance theory and psychological essentialism may provide a concerted account of representation and categorization across domains. PMID- 12749463 TI - Convex hull and tour crossings in the Euclidean traveling salesperson problem: implications for human performance studies. AB - Recently there has been growing interest among psychologists in human performance on the Euclidean traveling salesperson problem (E-TSP). A debate has been initiated on what strategy people use in solving visually presented E-TSP instances. The most prominent hypothesis is the convex-hull hypothesis, originally proposed by MacGregor and Ormerod (1996). We argue that, in the literature so far, there is no evidence for this hypothesis. Alternatively we propose and motivate the hypothesis that people aim at avoiding crossings. PMID- 12749465 TI - Effects of wording and stimulus format on the use of contingency information in causal judgment. AB - There are four kinds of contingency information: occurrences and nonoccurrences of an effect in the presence and in the absence of a cause. Previous studies have shown that these four kinds are not given equal weight in causal judgment. The present research was designed to test two hypotheses about this unequal weighting: that weightings are influenced by the form of the question and other features of the stimulus materials and that unequal weightings occur, in part, because individual differences in the use of contingency information are not evenly distributed across the four kinds of information. Support was found for both hypotheses. However, the effects of question wording were not always as had been predicted, indicating that more needs to be learned about how people interpret the task, instructions, and materials they are given. PMID- 12749464 TI - Does analogical transfer involve a term-to-term alignment? AB - According to dominant models of reasoning by analogy, analogical transfer requires subjects to first define a full one-to-one correspondence between the base and the target problems. Accordingly, these models predict that if there is a cross-mapping between the base and the target (cross-mapping exists when similar or identical elements in the base and the target play different roles), the time spent transferring a property from the base to the target will be greater. The present results are inconsistent with this prediction. Indeed, if the mapping task is more difficult in the cross-mapping condition than in the control condition, the time needed to make the transfer is not affected by the presence of a cross-mapping. Consequently, we conclude that the mapping phase is not a necessary condition for transfer. PMID- 12749466 TI - Is probability matching smart? Associations between probabilistic choices and cognitive ability. AB - In three experiments involving over 1,500 university students (n = 1,557) and two different probabilistic choice tasks, we found that the utility-maximizing strategy of choosing the most probable alternative was not the majority response. In a story problem version of a probabilistic choice task in which participants chose from among five different strategies,the maximizing response and the probability-matching response were each selected by a similar number of students (roughly 35% of the sample selected each). In a more continuous, or trial-by trial, task, the utility-maximizing response was chosen by only one half as many students asthe probability-matching response. More important, in both versions of the task, the participants preferring the utility-maximizing response were significantly higher in cognitive ability than were the participants showing a probability-matching tendency. Critiques of the traditional interpretation of probability matching as nonoptimal may well help explain why some humans are drawn to the nonmaximizing behavior of probability matching, but the traditional heuristics and biases interpretation can most easily accommodate the finding that participants high in computational ability are more likely to carry out the rule based cognitive procedures that lead to maximizing behavior. PMID- 12749467 TI - Egocentric organization of spatial activities in imagined navigation. AB - Studies on spatial frameworks suggest that the way we locate objects in imagined environments is influenced by the physical and functional properties of the world and our body. The present study provides evidence that such an influence also characterizes imagined navigation. In Experiment 1, participants followed spatial directions to construct an imagined path, while either keeping constant or updating their orientation at each step. A pattern of step times diagnostic of spatial frameworks was obtained in the updated-orientation but not in the constant-orientation condition. In Experiment 2, participants performed the updated-orientation condition with two levels of external support for the reference frame being used. Step times conformed to the predictions of spatial frameworks in both conditions. Both experiments also provided support that the processes involved in imagined navigation exhibit the operator-operand dynamics of other mental skills previously documented in the mental arithmetic domain. These results reinforce Piaget's (1954) notion that spatial displacements and integer arithmetic share a set of structural characteristics PMID- 12749468 TI - Is there a "strength effect" in automatic semantic priming? AB - According to spreading activation models of automatic priming, highly related associates should yield stronger priming effects than weakly related associates. The strength of relations is usually based on word association norms. However, this strength effect has been found in some studies but not in others. The present study suggests that one factor that might be responsible for this inconsistency is whether the weak associate is the primary or nonprimary response in the word association norms This possibility was explored by comparing priming effects of weak nonprimary and primary associates with those of strong primary associates. Comparable priming effects were found for the strong and weak primary associates whereas the weak nonprimary associates did not yield any priming effects. These results were obtained both in paired (Experiment 1) and single (Experiment 2) presentation priming procedures. Thus, the rank of the associate is an important factor in predicting the magnitude of the priming effect. PMID- 12749469 TI - All parts of an item are not equal: effects of phonological redundancy on immediate recall. AB - The process of redintegration is thought to use top-down knowledge to repair partly damaged memory traces. We explored redintegration in the immediate recall of lists from a limited pool of partly phonologically redundant pseudowords. In Experiment 1, four kinds of stimuli were created by adding the syllable /ne/ to two-syllable pseudowords, either to the middle (/tepa/ vs. /tenepa/) or to the end (/tepane/), or adding a different syllable to each item (/tepalo/, /vuropi/). The repeated syllable was thought to be available for redintegration. Lists of two-syllable pseudowords were recalled best, items with a redundant end were intermediate, and items with a redundant middle-syllable were as hard as nonredundant three-syllable items. In Experiment 2, the last syllable was predictable from context but not shared between all stimuli, reducing phonological similarity between items. Performance did not differ from the situation with identicallast syllables. In Experiment 3, a shared first syllable had a detrimental effecton memory. An error analysis showed that beneficial redundancy effects were accompanied by harmful similarity effects, impairing memory for nonredundant syllables. The balance between the two effects depended on syllable position. PMID- 12749470 TI - The effect of feature frequency on short-term recognition memory. AB - We report two experiments using Sternberg's (1969) multitrial recognition-memory paradigm. We used colored shapes as stimuli and manipulated the frequency of the shapes (but not of the colors) across trials. For lures containing an extralist shape (i.e., a shape not studied in the current study list), responses were faster if the shape had occurred infrequently than if it had occurred frequently in the preceding trials. For lures containing an extralist color and a studied shape, by contrast, the frequency of the shape in the preceding trials was irrelevant. We conclude that correct rejections depend solely on contradictory evidence. Furthermore, low-frequency target items were recognized more easily than high-frequency targets. Both the interaction of frequency with the features of the lures and the main effect of frequency for the targets are problematic for current accounts of recognition. PMID- 12749471 TI - Predicting and postdicting the effects of word frequency on memory. AB - In the experiments reported here, I replicate and extend recent results that reveal that judgments about the memorability of common and uncommon words differ qualitatively depending on whether they are made during study or elicited during a recognition test (Guttentag & Carroll, 1998). When assessing recognition ability for individual words, subjects predict superior performance for common words, but postdict better performance for uncommon words. This interaction suggests that subjects rely on different cues when making judgments during study than they do when making analogous judgments during the recognition test, and that the cues utilized during recognition lead judgments to be more accurate. The shift is then evident in later predictions Subjects who make postdictions consequently correctly predict superior recognition performance for uncommon words on a subsequent study list. When subjects are asked to make later predictions about recall performance, however, having made postdictions on a test of recognition does not mislead subjects into predicting superior recall performance for uncommon words. PMID- 12749472 TI - Incidental formation of episodic associations: the importance of sentential context. AB - The influence of relevant semantic context on the incidental formation of episodic associations between words was probed in two experiments. In Experiment 1, we examined the influence of associations formed incidentally between unrelated words presented either in isolation or embedded in a sentential context on subsequent explicit paired-associate leaning tested by cued recall. The results of Experiment 1 showed that the cued-recall rate of words studied in sentential context was higher than that of words co-occurring in isolated pairs. A subsequent single-items recognition test showed equal item memory for words studied in sentences than for words studied in isolated pairs, suggesting that the sentential context effect in cued recall indeed reflected stronger associations between paired words rather than better memory for single words. In Experiment 2, we ruled out memory for the entire sentence as an alternative explanation for the results of Experiment 1. We suggest two possible mechanisms to account for this advantage: First, pairs embedded in a sentence undergo semantic elaboration that might lead to the incidental formation of an association between them. Second, words embedded in a sentence enjoy the conjoint activation of compatible semantic features, a fact that may also facilitate the formation of an episodic association between them. The implications of these results for computational models using word representations based on co occurrence data are discussed. PMID- 12749473 TI - Themes, events, and episodes in autobiographical memory. AB - The process by which experience is divided into events was examined. Experiment 1 involved diarists recording their experiences over a 3-month period. Diary entries were later transcribed onto cards and the diarists arranged their cards so as to define events they had experienced, and in a separate phase arranged their cards so as to describe the themes that reflected their life. Examination of event- and theme-building strategies indicated that boundaries were frequently used, and events and themes were often formed from clusters of experience combined using content association rather than temporal sequence. Experiment 2 involved photographs taken by the participants, employed the same procedures as in Experiment 1, and revealed event- and theme-building strategies similar to those identified in Experiment 1. In Experiment 3 the size of the stimulus set from which events and themes were constructed was manipulated; this did not influence construction strategies. Overall, the experiments show that both autobiographical events and themes frequently consist of episodes taken from more than 1 day. PMID- 12749474 TI - Update: Severe acute respiratory syndrome--United States, 2003. AB - CDC continues to work with the World Health Organization (WHO) and other partners to investigate cases of severe acute respiratory syndrome (SARS). This report updates information on reported SARS cases worldwide and among U.S. residents and summarizes information on one additional case with laboratory evidence of infection with the SARS-associated coronavirus (SARS-CoV). PMID- 12749475 TI - Update: Adverse events following civilian smallpox vaccination--United States, 2003. AB - During January 24-April 18, 2003, smallpox vaccine was administered to 33,444 civilian health-care and public health workers in 54 jurisdictions to prepare the United States for a possible terrorist attack using smallpox virus. This report updates information on vaccine-associated adverse events among civilians vaccinated since the beginning of the program and among contacts of vaccinees, received by CDC from the Vaccine Adverse Event Reporting System (VAERS) as of April 18. PMID- 12749476 TI - Nationwide measles vaccination campaign for children aged 6 months-12 years- Afghanistan, 2002. AB - The public health infrastructure in Afghanistan has been devastated by 23 years of civil war, and both the infant mortality rate (165 per 1,000 live-born infants) and the mortality rate for children aged <5 years (256 per 1,000 live born infants) are among the highest in the world. The major causes of death among children aged <10 years are diarrhea (32%), measles (25%), respiratory tract infections (13%), and other causes (30%), including malnutrition, scurvy, chronic diseases, and fever of unknown origin. Measles accounts for an estimated 30,000 35,000 deaths each year in Afghanistan. To reduce measles-related mortality, during 2002, the Ministry of Health (MoH) of the Interim Government of Afghanistan, with the support of international organizations, organized a nationwide measles vaccination campaign for children aged 6 months-12 years. This report describes the planning, implementation, and impact of this campaign. The findings suggest that the campaign had a major impact on reducing measles-related mortality. Similar campaigns might be feasible in countries affected by complex emergencies. PMID- 12749477 TI - Progress toward global eradication of poliomyelitis, 2002. AB - Since the 1988 World Health Assembly resolution to eradicate poliomyelitis globally through 2002, the number of countries where polio is endemic declined from 125 to seven, and the estimated incidence of polio decreased >99%. In 2002, the European Region became the third World Health Organization (WHO) region certified as polio-free, joining the Region of the Americas and the Western Pacific Region, certified polio-free in 1994 and 2000, respectively. Despite these achievements, a provisional total of 1,920 polio cases were reported during 2002, a substantial increase from 483 in 2001, reflecting primarily the large polio epidemic in India. This report summarizes global progress achieved in polio eradication during 2002 and describes remaining challenges. PMID- 12749478 TI - Insecticide-treated bed nets. PMID- 12749479 TI - The efficacy of permethrin-treated bed nets on child mortality and morbidity in western Kenya I. Development of infrastructure and description of study site. AB - Randomized controlled trials in sub-Saharan Africa have shown that permethrin treated bed nets and curtains reduce all-cause child mortality by 15-33% in areas with low or high but seasonal malaria transmission. This report describes the study site for a community-based, group-randomized, controlled trial in an area of high and year-round malaria transmission in western Kenya. We outline the development of the human and physical infrastructure required to conduct this trial and discuss some of the difficulties encountered and lessons learned in conducting it. PMID- 12749480 TI - The efficacy of permethrin-treated bed nets on child mortality and morbidity in western Kenya II. Study design and methods. AB - This paper describes the study design and methods used in a large community based, group-randomized, controlled trial of permethrin-treated bed nets (ITNs) in an area with intense, perennial malaria transmission in western Kenya conducted between 1996 and 1999. A multi-disciplinary framework was used to explore the efficacy of ITNs in the reduction of all-cause mortality in children less than five years old, the clinical, entomologic, immunologic, and economic impact of ITNs, the social and behavioral determinants of ITN use, and the use of a geographic information system to allow for spatial analyses of these outcomes. Methodologic difficulties encountered in such large-scale field trials are discussed. PMID- 12749481 TI - Impact of permethrin-treated bed nets on entomologic indices in an area of intense year-round malaria transmission. AB - The effect of permethrin-treated bed nets (ITNs) on malaria vectors was studied as part of a large-scale, randomized, controlled trial in western Kenya. Indoor resting densities of fed Anopheles gambiae s.l. and An. funestus in intervention houses were 58.5% (P = 0.010) and 94.5% (P = 0.001) lower, respectively, compared with control houses. The sporozoite infection rate in An. gambiae s.l. was 0.8% in intervention areas compared with 3.4% (P = 0.026) in control areas, while the sporozoite infection rates in An. funestus were not significantly different between the two areas. We estimated the overall transmission of Plasmodium falciparum in intervention areas to be 90% lower than in control areas. Permethrin resistance was not detected during the study period. As measured by densities of An. gambiae s.l., the efficacy of bed nets decreased if one or more residents did not sleep under a net or if bed nets had not been re-treated within six months. These results indicate that ITNs are optimally effective if used every night and if permethrin is reapplied at least biannually. PMID- 12749482 TI - Efficacy of permethrin-treated bed nets in the prevention of mortality in young children in an area of high perennial malaria transmission in western Kenya. AB - A group-randomized controlled trial of insecticide (permethrin)-treated bed nets (ITNs) was conducted in an area of high perennial malaria transmission in western Kenya to test the effect of ITNs on all-cause mortality in children 1-59 months of age. Child deaths were monitored over a two-year period by biannual household census in Asembo (1997-1998) and in Gem (1998-1999). Overall, 1,722 deaths occurred in children 1-59 months followed for 35,932 child-years. Crude mortality rates/1,000 child-years were 51.9 versus 43.9 in control and ITN villages in children 1-59 months old. The protective efficacy (PE) (95% confidence interval) adjusted for age, study year, study site, and season was 16% (6-25%). Corresponding figures in 1-11- and 12-59-month-old children in control and ITN villages were 133.3 versus 102.3, PE = 23% (11-34%) and 31.1 versus 28.7, PE = 7% (-6-19%). The numbers of lives saved/1,000 child-years were 8, 31, and 2 for the groups 1-59, 1-11, and 12-59 months old, respectively. Stratified analysis by time to insecticide re-treatment showed that the PE of ITNs re-treated per study protocol (every six months) was 20% (10-29%), overall and 26% (12-37%) and 14% ( 1-26%) in 1-11- and 12-59-month-old children, respectively. ITNs prevent approximately one in four infant deaths in areas of intense perennial malaria transmission, but their efficacy is compromised if re-treatment is delayed beyond six months. PMID- 12749483 TI - Comparison of government statistics and demographic surveillance to monitor mortality in children less than five years old in rural western Kenya. AB - Estimates of mortality in children less than five years old using government civil registration statistics (passive surveillance) were compared against statistics generated by active demographic surveillance during a randomized controlled trial of permethrin-treated bed nets (ITNs) in western Kenya. Mortality rates were two-fold lower when estimated through civil registration compared with active prospective surveillance (rate ratio [RR] = 0.51, 95% confidence interval [CI] = 0.44-0.59). While civil registration underestimated deaths, particularly in the neonatal period, the age distribution of deaths in children 1-59 months of age was the same as with active surveillance. Seasonal mortality trends were also similar. There was no agreement between cause of death recorded by active and passive surveillance. Verbal autopsy estimated that half of all deaths were associated with malaria and pneumonia, but civil registration markedly under-reported these illnesses; incidence RR (95% CI) = 0.18 (0.14 0.24), and 0.05 (0.03-0.08), respectively, while over-reporting deaths due to measles (RR = 15.5 [95% CI = 7.3-33.2]). Government statistics under-represent mortality, particularly neonatal mortality, in children less than five years of age in rural areas of Kenya. They can provide accurate information on the age distribution of deaths among children 1-59 months old, and on seasonal trends, but not on disease-specific mortality. PMID- 12749485 TI - Diagnostic and prescribing practices in peripheral health facilities in rural western Kenya. AB - Health facility ledgers of 11 rural health facilities in western Kenya were reviewed to evaluate diagnostic and prescribing practices. Clinics lacked laboratory facilities. Of 14,267 sick child visits (SCVs), 76% were diagnosed with malaria and/or upper respiratory infections. Other diagnoses were recorded in less than 5% of SCVs. Although two-thirds of malaria cases were diagnosed with co-infections, less than 3% were concomitantly diagnosed with anemia. Chloroquine and penicillin constituted 94% of prescriptions. Half of children given a sole diagnosis of measles or pneumonia were prescribed chloroquine, and 22% of children with a sole diagnosis of malaria were given penicillin. Antimalarials other than chloroquine were rarely prescribed. Only 12% of children diagnosed with anemia were prescribed iron supplementation, while 53% received folic acid. This study highlights limited diagnostic and prescribing practices and a lack of adherence to national treatment guidelines in rural western Kenya. PMID- 12749484 TI - Impact of permethrin-treated bed nets on the incidence of sick child visits to peripheral health facilities. AB - During a randomized controlled trial of insecticide (permethrin)-treated bed nets (ITNs) in an area with intense malaria transmission in western Kenya, we monitored 20,915 sick child visits (SCVs) by children less than five years of age visiting seven peripheral health facilities. The SCVs were monitored over a four year period both before (1995-1996) and during the intervention (1997-1998). Results are used to estimate the effect of ITNs on the burden of malaria in this community and to evaluate the potential role of these facilities in assessment of the impact of large-scale public health interventions. Compared with baseline, a 27% greater reduction in the incidence of SCVs was seen in ITN villages than in control villages (37% versus 10%; P = 0.049). A similar reduction was observed in SCVs diagnosed as malaria (35% reduction in ITN villages versus 5% reduction in controls; P = 0.04). Two-hundred sixteen SCVs per 1,000 child-years were prevented; three-fourths of these were in children less than 24 months old. As a consequence of lack of laboratory facilities, severe anemia was rarely (< 2%) diagnosed, regardless of intervention status. No effect of ITNs on the incidence of respiratory tract infections, diarrhea, and other commonly diagnosed childhood illnesses was observed. The ITNs reduced the number of SCVs due to malaria, but had no effect on other illnesses. Routine statistics from these facilities provided useful information on trends in malaria incidence, but underestimated the burden of severe anemia. PMID- 12749486 TI - Reduction of malaria during pregnancy by permethrin-treated bed nets in an area of intense perennial malaria transmission in western Kenya. AB - The impact of insecticide (permethrin)-treated bed nets (ITNs) on malaria in pregnancy was studied in a rural area in western Kenya with intense perennial malaria transmission. All households in 40 of 79 villages were randomized to receive ITNs by January 1997. The ITNs were distributed in control villages two years later. Complete data on birth outcome were available on 2,754 (89.6%) of 3,072 deliveries. Women (n = 780) were followed monthly throughout pregnancy in 19 of 79 villages. Among gravidae 1-4, ITNs were associated with reductions of 38% (95% confidence interval [CI] = 17-54%) in the incidence of malaria parasitemia and 47% (95% CI = 6-71%) in the incidence of severe malarial anemia (hemoglobin level < 8 g/dL with parasitemia) during pregnancy. At the time of delivery, mean hemoglobin levels were 0.6 g/dL (95% CI = 0.01-1.2 g/dL) higher, the prevalence of placental or maternal malaria was reduced by 35% (95% CI = 20 47%), and the prevalence of low birth weight was reduced by 28% (95% CI = 2-47%) in gravidae 1-4 from ITN villages. No beneficial impact was observed in gravidae five or higher. In areas of intense perennial malaria transmission, permethrin treated bed nets reduce the adverse effect of malaria during the first four pregnancies. PMID- 12749487 TI - Effects of permethrin-treated bed nets on immunity to malaria in western Kenya I. Antibody responses in pregnant women and cord blood in an area of intense malaria transmission. AB - As part of a community-based group-randomized trial on the impact of permethrin treated bed nets (ITNs) on malaria in pregnancy in a holoendemic area of western Kenya, we assessed their effects on antibody responses to Plasmodium falciparum pre-erythrocytic antigens (recombinant circumsporozoite protein [CSP] and peptides complimentary to the repeat region of the liver stage antigen-1 [LSA-1]) and blood stage antigen (recombinant C-terminal domain of the merozoite surface protein-1 [MSP-1(19) kD]) in paired maternal/cord plasma samples obtained from 296 deliveries (157 from ITN villages and 139 control villages). Levels of total IgG and IgG subclasses 1-3 to LSA-1 and total IgG and IgG3 to MSP-1 were lower, whereas those of total IgG to CSP were significantly higher in women from ITN villages than those from control villages. In cord plasma, levels of total IgG and IgG2 to LSA-1 and IgG3 to MSP-1 were lower in ITN villages than in control villages, but antibody responses to CSP were similar. Our results suggest that the use of ITNs decreases antibody responses to LSA-1 and MSP-1 antigens in pregnant women with associated reductions in levels of the same antibodies in cord blood. In contrast, ITN use was found to be associated with increased antibody responses to CSP in pregnant women, but had no effect on antibody levels to CSP in cord blood. PMID- 12749488 TI - Impact of permethrin-treated bed nets on malaria, anemia, and growth in infants in an area of intense perennial malaria transmission in western Kenya. AB - As part of a community-based, group-randomized, controlled trial of insecticide treated bed nets (ITNs) in an area with intense malaria transmission in western Kenya, a birth cohort (n = 833) was followed monthly until the age of 24 months to determine the potential beneficial and adverse effects of reduced malaria exposure during pregnancy and infancy. Malaria transmission and morbidity were comparable pre-intervention. The ITNs reduced malaria attack rates (force of infection) in infancy by 74%, and delayed the median time-to-first parasitemia (4.5 to 10.7 months; P < 0.0001). The incidence of both clinical malaria and moderate-severe anemia (hemoglobin level <7 g/dL) were reduced by 60% (P < 0.001 for both). Protective efficacy was greatest in infants less than three months old and similar in older infants and one-year-old children. Efficacy was lowest in the dry season. Infants from ITN villages experienced better height and weight gain. In areas of intense perennial malaria transmission, ITNs substantially reduce exposure to malaria and subsequent malaria-associated morbidity in children less than 24 months old. Reduced malaria exposure during infancy did not result, with continued ITN use, in increased malaria morbidity in one-year-old children. PMID- 12749489 TI - Impact of permethrin-treated bed nets on growth, nutritional status, and body composition of primary school children in western Kenya. AB - Insecticide-treated bed nets (ITNs) have been demonstrated to reduce morbidity and mortality in children less than five years of age. They have also been shown to improve the nutritional status of these children, but little is known about their impact on the nutritional status of school-age children. We evaluated the impact of ITNs on growth, nutritional status, and body composition of primary schoolchildren less than 13 years of age living in an area of intense perennial malaria transmission in western Kenya. The ITNs did not have a significant impact on linear growth or summary measures of protein-energy malnutrition in this age group. This lack of efficacy most likely relates to the reduced burden of malaria in this age group in a setting of stable transmission pressure. Use of ITNs was associated with a change in body composition with an increase in percent lean body mass (1.2%; P = 0.04). This may be consequent to reduced exposure to malaria with subsequent reduced elaboration of pro-inflammatory cytokines known to promote muscle wasting. PMID- 12749490 TI - Permethrin-treated bed nets in the prevention of malaria and anemia in adolescent schoolgirls in western Kenya. AB - The impact of insecticide (permethrin)-treated bed nets (ITNs) on the health of adolescent schoolgirls was investigated during a community-based, randomized, controlled trial of ITNs in western Kenya. Two school-based cross-sectional surveys were conducted to determine the prevalence of malaria and anemia in 644 schoolgirls 12-18 years old in a rural area with intense perennial malaria transmission. In 12- and 13-year-old schoolgirls, ITNs were associated with a reduced prevalence of all cause anemia (hemoglobin level <12 g/dL, 16.9% versus 31.4%, adjusted odds ratio [OR] = 0.38, 95% confidence interval [CI] = 0.21, 0.69%) and a 0.34 g/dL (95% CI = 0.02, 0.66) increase in mean hemoglobin concentrations. No beneficial effect on all-cause anemia (adjusted OR = 0.79, 95% CI = 0.43, 1.45) or hemoglobin concentrations (difference in mean = 0.14 g/dL, 95% CI = -0.24, 0.53) was evident in older girls. In all age groups, no effect was found on malaria parasite prevalence or density, clinical malaria, all-cause morbidity, standard measures of nutritional status and growth, or the use of antimalarials and other medications. ITNs approximately halved the prevalence of mild anemia in young, school-attending, non-pregnant, adolescent girls, but had no impact in older girls or on other malaria-associated morbidity or nutritional status. PMID- 12749492 TI - Impact of permethrin-treated bed nets on malaria and all-cause morbidity in young children in an area of intense perennial malaria transmission in western Kenya: cross-sectional survey. AB - Information on the impact of insecticide (permethrin)-treated bed nets (ITNs) from randomized controlled trials in areas of intense perennial malaria transmission is limited. As part of a large-scale, community-based, group randomized controlled trial of the effect of ITNs on childhood mortality in a holoendemic area in western Kenya, we conducted three cross-sectional surveys in 60 villages to assess the impact of ITNs on morbidity in 1,890 children less than three years old. Children in ITN and control villages were comparable pre intervention, but after the introduction of ITNs, children in intervention villages were less likely to have recently experienced illness requiring treatment (protective efficacy [95% confidence intervals] = 15% [1-26%]), have an enlarged spleen (32% [20-43%]), be parasitemic (19% [11-27%]), have clinical malaria (44% [6-66%]), have moderately severe anemia (hemoglobin level < 7.0 g/dL; 39% [18-54%]), or have a pruritic body rash, presumably from reduced nuisance insect bites (38% [24-50%]). Use of ITNs was also associated with significantly higher mean weight-for-age Z-scores and mid-upper arm circumferences. There was no evidence, however, that ITNs reduced the risk of helminth infections, diarrhea, or upper or lower respiratory tract infections. The ITNs substantially reduced malaria-associated morbidity and improved weight gain in young children in this area of intense perennial malaria transmission. PMID- 12749491 TI - Prevalence and severity of malnutrition in pre-school children in a rural area of western Kenya. AB - We determined the nutritional status of children less than five years of age in an area in rural western Kenya with intense malaria transmission, a high prevalence of severe anemia and human immunodeficiency virus, and high infant and under-five mortality (176/1,000 and 259/1,000). No information is available on the prevalence of malnutrition in this area. Three cross-sectional surveys were conducted between 1996 and 1998 to monitor the effect of insecticide-treated bed nets on child morbidity. Anthropometric indices are presented for 2,103 children collected prior to and during intervention (controls only). The prevalence of stunting (Z-scores for height-for-age [HAZ] <-2), wasting (Z-scores for weight for-height [WHZ] <-2) and being underweight (Z-scores for weight-for-age [WAZ] < 2) was 30%, 4%, and 20%, respectively. This was severe (Z-score <-3) in 12% (stunting), 1% (wasting), and 5% (underweight) of the children. Few children less than three months of age were malnourished (<2%), but height-for-age and weight forage deficits increased rapidly in children 3-18 months of age, and were greatest in children 18-23 months old (44% stunted and 34% underweight). While the mean HAZ and WAZ stabilized from 24 months of age onwards, they still remained substantially below the reference median with no evidence of catch-up growth. Malnutrition is likely to interact with infectious diseases, placing children 3-24 months of age at high risk of premature death in this area. PMID- 12749493 TI - Effects of permethrin-treated bed nets on immunity to malaria in western Kenya II. Antibody responses in young children in an area of intense malaria transmission. AB - As part of a large community-based trial on the impact of insecticide (permethrin)-treated bed nets (ITNs) on childhood morbidity and mortality in an area of intense perennial malaria transmission in western Kenya, we assessed the effects of ITNs on malaria-specific humoral responses in young children. The IgG responses to Plasmodium falciparum pre-erythrocytic antigens circumsporozoite protein (CSP) and liver stage antigen-1 (LSA-1) and the blood stage antigen merozoite surface protein-1 (MSP-1(19) kD) in children less than three years old were investigated during a series of cross-sectional surveys. At 14 and 22 months after the introduction of ITNs, the frequencies and levels of IgG to CSP and LSA 1 were significantly lower in children from ITN villages than in children from control villages (P < 0.001). In contrast, the prevalence of IgG to MSP-1 was significantly higher in children from ITN villages at 14 months (P = 0.0069), but not at 22 months. Our results show that decreased exposure by ITNs reduces IgG responses to pre-erythrocytic antigens, but there was no evidence that two years of ITN use compromises IgG responses to blood stage antigens in these young children in this malaria holoendemic area. PMID- 12749494 TI - Effect of permethrin-treated bed nets on the spatial distribution of malaria vectors in western Kenya. AB - The effect of insecticide (permethrin)-treated bed nets (ITNs) on the spatial distribution of malaria vectors in neighboring villages lacking ITNs was studied during a randomized controlled trial of ITNs in western Kenya. There was a trend of decreased abundance of Anopheles gambiae with decreasing distance from intervention villages both before (P = 0.027) and after (P = 0.002) introduction of ITNs, but this trend was significantly stronger after ITNs were introduced (P = 0.05). For An. funestus, no pre-intervention trend was observed (P = 0.373), but after the intervention, a trend of decreased abundance with closer proximity to intervention compounds developed (P = 0.027). Reduction in mosquito populations in villages lacking ITNs was most apparent in compounds located within 600 meters of intervention villages. Sporozoite infection rates decreased in control areas following the introduction of ITNs (P < 0.001 for both species), but no spatial association was detected between sporozoite rates and distance to nearest intervention village. We conclude that high coverage of ITNs is associated with a community-wide suppression of mosquito populations that is detectable in neighboring villages lacking ITNs, thereby affording individuals residing in these villages some protection against malaria. PMID- 12749495 TI - Community-wide effects of permethrin-treated bed nets on child mortality and malaria morbidity in western Kenya. AB - Spatial analyses of the effect of insecticide (permethrin)-treated bed nets (ITNs) on nearby households both with and without ITNs was performed in the context of a large-scale, group-randomized, controlled mortality trial in Asembo, western Kenya. Results illustrate a protective effect of ITNs on compounds lacking ITNs located within 300 meters of compounds with ITNs for child mortality, moderate anemia, high-density parasitemia, and hemoglobin levels. This community effect on nearby compounds without nets is approximately as strong as the effect observed within villages with ITNs. This implies that in areas with intense malaria transmission with high ITN coverage, the primary effect of insecticide-treated nets is via area-wide effects on the mosquito population and not, as commonly supposed, by simple imposition of a physical barrier protecting individuals from biting. The strength of the community effect depended upon the proportion of nearby compounds with treated nets. To maximize their public health impact, high coverage with treated nets is essential. PMID- 12749496 TI - Community reactions to the introduction of permethrin-treated bed nets for malaria control during a randomized controlled trial in western Kenya. AB - Prior to implementation of a randomized controlled trial of insecticide (permethrin)-treated bed nets (ITNs) in western Kenya, ethnographic studies were conducted to understand local perceptions of disease, sleeping patterns, and other factors that might affect use of ITNs. Educational activities took place prior to distribution, but immediately after distribution in Asembo only approximately half of the ITNs were in use. A qualitative study was then conducted to identify the community's perceptions about ITNs and the ITN project. While participants ranked malaria as important and recognized that malaria prevention could be beneficial, they believed ITNs would be only partly effective due to the perception that malaria has multiple causes. Concerns expressed included fear of the insecticide, thought by some to be a toxic family planning aid, the taking of blood during clinical studies, and the mixing up of family ITNs during net re-treatment, which would violate cultural taboos. Attempts were made to allay fears by improved communication on these subjects and modification of the study design. PMID- 12749497 TI - Factors affecting use of permethrin-treated bed nets during a randomized controlled trial in western Kenya. AB - Adherence with permethrin-treated bed net (ITN) use and their proper deployment was directly observed in 2,178 individuals (784 households) participating in a large-scale trial of ITNs on child mortality in western Kenya. The ITNs were distributed free of charge to ensure high coverage, resulting in a ratio of 1.46 persons per ITN. Approximately 30% of ITNs present were unused. The overall percentage adherence was 72.3%. The probability of adherence by individuals depended strongly on age (relative risk [RR] = 0.86, 95% confidence limit [CL] = 0.78-0.94), in which children less than five years of age were less likely to use ITNs than older individuals, and temperature, in which ITNs were more likely to be used in periods of cooler weather. A marginally significant diminution in adherence during the second year of the project was also observed (RR = 0.83, 95% CL = 0.68-1.01). Mosquito numbers, relative wealth, number of house occupants, and the educational level of the head of the household had no effect on adherence. In unstructured questioning of house residents, excessive heat was often cited as a reason for not deploying the child's ITN. The most important reason for non-adherence was disruption of sleeping arrangements, indicating that ITNs were not readily redeployed in the face of shifting sleeping patterns due to visitors, funerals, house construction, and other events. Challenges faced by health education programs to maximize adherence with ITN use are discussed. PMID- 12749498 TI - Perceptions of bed nets and malaria prevention before and after a randomized controlled trial of permethrin-treated bed nets in western Kenya. AB - A study of mothers' perceptions regarding bed nets and malaria was conducted before and after a randomized controlled trial of insecticide (permethrin) treated bed nets (ITNs) in western Kenya. Awareness about the trial and the rationale for bed net use increased by the end of the trial. Knowledge that mosquitoes caused malaria also increased; however, a higher proportion of mothers from control, rather than intervention villages, cited this (44.4% versus 27.9%; P < 0.001). Mothers from intervention villages were more knowledgeable about the use and maintenance of bed nets and re-treatment with insecticide. Both groups specified advantages of ITNs. Mothers from intervention villages noted practical advantages such as protection against bedbugs and falling roof debris. Few (< 1%) mothers indicated that ITNs protected children against malaria. Intervention homes used significantly fewer mosquito coils, insect spray, medicines, and burned cow dung less often compared with those in control villages. Mothers were willing to pay approximately 4.5 U.S. dollars for a regular bed net, but only 10.5 U.S. cents (intervention) and 0.036 (control) for re-treating a bed net. This study suggests that, despite two years of experience of use, bed nets and insecticides would not be purchased as a household priority in this impoverished rural community. PMID- 12749499 TI - The household-level economics of using permethrin-treated bed nets to prevent malaria in children less than five years of age. AB - We measured the two-week household-level economic impact of insecticide (permethrin)-treated bed nets (ITNs) used to prevent malaria among children less than five years of age in Asembo, Kenya. The ITNs induced a two-week reduction of 15 Kenyan shillings (KSH) (0.25 U.S. dollars; P < 0.0001) in health care expenditures, but a statistically insignificant 0.5 day (P = 0.280) reduction in household time lost due to caring for sick children. The equivalent annual threshold cost was estimated at 6.50 U.S. dollars (95% confidence interval = 3.12 9.86). If the actual purchase price and maintenance costs of ITNs were greater than this threshold, then households would pay more than they would save (and vice-versa). Both seasonal effects and number of children per household had larger impacts than ITNs on health care expenditures and time lost from household activities. Health care expenditures by a household without ITNs and one child were only 32 KSH per two weeks (0.50 U.S. dollars; P = 0.002), leaving little opportunity for household-level, ITN-induced direct savings. The widespread adoption of the ITNs will therefore probably require a subsidy. PMID- 12749500 TI - The cost-effectiveness of permethrin-treated bed nets in an area of intense malaria transmission in western Kenya. AB - This study compared the costs and effects of insecticide (permethrin)-treated bed net (ITN) use in children less than five years of age in an area of intense, perennial malaria transmission in western Kenya. The data were derived from a group-randomized controlled trial of ITNs conducted between 1996 and 1999. The annual net cost per life-year gained was 34 U.S. dollars and the net annual cost per all-cause sick child clinic visit averted was 49 U.S. dollars. After taking into account a community effect (protection from malaria afforded to non-ITN users who lived within 300 meters from users) these estimates decreased to 25 U.S. dollars and 38 U.S. dollars, respectively. This study provides further evidence that ITNs are a highly cost-effective use of scarce health care resources. PMID- 12749502 TI - A review of the effects of almotriptan and other triptans on clinical trial outcomes that are meaningful to patients with migraine. AB - BACKGROUND: Traditional end points in clinical trials of migraine therapy, such as 2-hour pain response, may not fully address the outcomes patients consider most important: rapid and sustained freedom from pain over 24 hours, and a low, placebo-like incidence of adverse events. A composite efficacy measure such as the sustained pain-free rate (no pain by 2 hours after dosing, no recurrence, no use of rescue medication from 2 to 24 hours after dosing) may be more appropriate. OBJECTIVE: Clinically relevant differences between almotriptan and other triptans were reviewed in the context of the attributes of acute migraine treatment that patients consider most important. METHODS: This review was based on published reports of open-label and placebo-controlled clinical trials of almotriptan, results of a survey concerning the attributes patients consider most important in a migraine medication, and a published meta-analysis of 53 placebo controlled clinical trials of triptans involving >24,000 patients. RESULTS: Almotriptan was effective and well tolerated in the placebo-controlled clinical trials; results of the 6- and 12-month open-label studies supported its good tolerability profile. A respective 87% and 86% of respondents to the patient survey indicated that they considered complete freedom from pain and no recurrence among the most important attributes of migraine treatment, both of which are included in the sustained pain-free rate. In the meta-analysis, almotriptan had a favorable efficacy and tolerability profile compared with other triptans, particularly with respect to sustained pain-free rate, which was significantly higher with almotriptan 12.5 mg compared with sumatriptan 100 mg (25.9% vs 20.0%, respectively; P < 0.05). In addition, the placebo-subtracted rate of adverse events was significantly lower with almotriptan compared with sumatriptan (1.8% vs 4.4%, respectively; P < 0.05). Results of a head-to-head placebo-controlled trial of almotriptan 12.5 mg and sumatriptan 100 mg supported the balance of efficacy and tolerability observed for almotriptan in the meta analysis. CONCLUSIONS: Data from clinical trials suggest that almotriptan is effective and well tolerated in the treatment of acute migraine pain. Based on a sustained pain-free rate that is among the highest and an adverse-event rate that is among the lowest for the triptans, almotriptan represents a therapeutic option for the initial treatment of acute migraine with or without aura. PMID- 12749501 TI - Implications of the western Kenya permethrin-treated bed net study for policy, program implementation, and future research. AB - The fifth, and probably last, large-scale, group-randomized, controlled trial of insecticide (permethrin)-treated bed nets (ITNs) showed that ITNs are efficacious in reducing all-cause post-neonatal mortality in an area of intense, perennial malaria transmission. The trial helped to define pregnant women and infants as target groups for this intervention in high transmission settings. High population coverage with ITNs in both target and non-target groups may be critical to enhance health and survival in pregnant women and infants. The proportion of households with ITNs (coverage), the proportion of individuals properly deploying ITNs each night (adherence), and the proportion of nets properly treated with insecticide (treatment) are the three key determinants of effectiveness of large-scale ITN programs. These three simple outcomes should serve as the basis for program objectives and monitoring and evaluation efforts. Coverage effects and economic analysis support the proposition that ITNs may be viewed as a public good, worthy of public support. Research should continue to improve the intervention tools (the net, the insecticide, and methods for durable treatment and re-treatment) and their deployment. PMID- 12749503 TI - Thalidomide: a review of approved and investigational uses. AB - BACKGROUND: Thalidomide is best known as a major teratogen that caused birth defects in up to 12,000 children in the 1960s. More recently, this agent has been approved by the US Food and Drug Administration for the treatment of erythema nodosum leprosum (ENL) through a restricted-use program. Its immunomodulatory, anti-inflammatory, and antiangiogenic properties are currently under study in a number of clinical conditions. OBJECTIVE: This article reviews the pharmacology of thalidomide; its approved and off-label uses in dermatologic, oncologic, and gastrointestinal conditions; and adverse events associated with its use. METHODS: Relevant articles were identified through searches of MEDLINE (1966-June 2002), International Pharmaceutical Abstracts (1970-June 2002), and EMBASE (1990-June 2002). Search terms included but were not limited to thalidomide, pharmacokinetics, pharmacology, therapeutic use, and teratogenicity, as well as terms for specific disease states and adverse events. Further publications were identified from the reference lists of the reviewed articles. Abstracts of recent symposia were obtained from the American Society of Clinical Oncology Web site. RESULTS: Thalidomide is thought to exert its therapeutic effect through the modulation of cytokines, particularly tumor necrosis factor-alpha. In addition to its approved indication for ENL, thalidomide has been studied in various other conditions, including graft-versus-host disease, discoid lupus erythematosus, sarcoidosis, relapsed/refractory multiple myeloma, Waldenstrom's macroglobulinemia, myelodysplastic syndromes, acute myeloid leukemia, myelofibrosis with myeloid metaplasia, renal cell carcinoma, malignant gliomas, prostate cancer, Kaposi's sarcoma, colorectal carcinoma, oral aphthous ulcers, Behcet's disease, Crohn's disease, and HIV/AIDS-associated wasting. Adverse events most frequently associated with its use include somnolence, constipation, rash, peripheral neuropathy, and thromboembolism. CONCLUSIONS: Use of thalidomide is limited by toxicity, limited efficacy data, and restricted access. Evidence of its efficacy in conditions other than ENL awaits the results of controlled clinical trials. PMID- 12749504 TI - Drotrecogin alfa (recombinant human activated protein C) for the treatment of severe sepsis. AB - BACKGROUND: The search for a life-preserving drug to treat sepsis has increased understanding of the pathogenesis of the process but produced little in the way of successful treatments. The prospective, randomized, double-blind, placebo controlled, Phase III, multicenter Recombinant Human Activated Protein C Worldwide Evaluation in Severe Sepsis (PROWESS) trial suggested that drotrecogin alfa--recombinant human activated protein C--significantly improved 28-day mortality rates in acute sepsis (P = 0.005). OBJECTIVES: The goals of this drug review were to summarize the recent findings regarding the pathogenesis of sepsis and septic shock, as well as the results of select immunomodulator drug trials, and to offer a comprehensive review of the mechanism of action, pharmacokinetic profile, efficacy and safety profile, and pharmacoeconomics of drotrecogin alfa. METHODS: The English-language literature was searched using the EMBASE and MEDLINE databases. In EMBASE, the subject headings drotrecogin, activated protein C, and sepsis were used to search publications from 1980 through September 2002. In MEDLINE, the MeSH heading protein C and subject heading sepsis were used to search publications from 1966 through September 2002. Published abstracts of recent meetings and proceedings of the US Food and Drug Administration were also reviewed. RESULTS: Drotrecogin alfa mimics the endogenous protein depleted during acute sepsis. Its activity as an antithrombotic, anti-inflammatory, and profibrinolytic agent appears to diminish the negative outcomes of acute sepsis, notably mortality at 28 days. The results of the PROWESS trial support this finding. A bleeding risk was noted during Phase II and III trials despite efforts to exclude those patients at high risk of bleeding. CONCLUSIONS: Drotrecogin alfa is the first adjunctive agent for the treatment of sepsis to display clinically and statistically significant effects on mortality rates at 28 days. Many questions remain regarding which patients are ideal candidates for treatment. New research and treatment guidelines are necessary to address these questions. PMID- 12749505 TI - Efficacy and tolerability of 5-day, once-daily telithromycin compared with 10 day, twice-daily clarithromycin for the treatment of group A beta-hemolytic streptococcal tonsillitis/pharyngitis: a multicenter, randomized, double-blind, parallel-group study. AB - BACKGROUND: Telithromycin, a ketolide antibacterial, has been developed for the treatment of community-acquired respiratory infections. OBJECTIVE: This study compared the efficacy and tolerability of 5-day, once-daily telithromycin with 10 day, twice-daily clarithromycin in adolescents and adults with acute tonsillitis/pharyngitis caused by group A beta-hemolytic streptococci ([GABHS] Streptococcus pyogenes). METHODS: In this multicenter, randomized, double-blind, parallel-group study, adolescent (aged > or = 13 years) and adult patients with a diagnosis of GABHS tonsillitis/pharyngitis received once-daily telithromycin 800 mg for 5 days (followed by placebo for 5 days) or twice-daily clarithromycin 250 mg for 10 days. Bacteriologic and clinical outcomes were assessed at a test-of cure visit (days 16 to 23) and a late posttherapy visit (days 31 to 45). RESULTS: A total of 526 patients were enrolled in the study, of which 463 (288 females, 175 males) were randomized to receive treatment (telithromycin, n = 232; clarithromycin, n = 231). The mean age of the telithromycin group was 30.9 years; in the clarithromycin group, it was 30.0 years. Bacterial eradication was achieved in 91.3% of telithromycin-treated patients and 88.1% of clarithromycin recipients (difference, 3.2%; 95% CI, -4.5 to 11.0). Clinical cure was achieved in 92.7% of telithromycin recipients and 91.1% of clarithromycin-treated patients (difference, 1.6%; 95% CI, -5.5 to 8.6). Bacteriologic and clinical cures for the 2 treatment groups also were similar at the late posttherapy visit. Treatment related adverse events occurred more frequently in the telithromycin group than the clarithromycin group (67.2% vs 57.5%, respectively); diarrhea, nausea, and vomiting were significantly more common with telithromycin than with clarithromycin (P = 0.004, 0.010, and 0.001, respectively). Adverse events were generally mild. CONCLUSION: This study demonstrates that telithromycin 800 mg once daily for 5 days was an effective and generally well-tolerated treatment for tonsillitis/pharyngitis caused by GABHS, providing similar bacteriologic and clinical efficacy to clarithromycin 250 mg twice daily for 10 days in the per protocol population. PMID- 12749506 TI - A multicenter, randomized, double-blind, double-dummy, placebo- and active controlled, parallel-group comparison of diclofenac-K and ibuprofen for the treatment of adults with influenza-like symptoms. AB - BACKGROUND: The alleviation of influenza-like symptoms, such as fever, headache, and muscle/joint aches and pains, is important so that sufferers can return to their normal daily activities. A flexible dosing regimen is proposed, starting with an initial dose of 2 tablets (2 x 12.5 mg), followed by 1 to 2 tablets every 4 to 6 hours as needed, to a maximum daily dose of 75 mg for up to 3 days for fever and 5 days for pain. This flexible dosing regimen matches the existing over the-counter dosing regimen of ibuprofen, which allows the patient to adjust the treatment according to the type, duration, and severity of symptoms. OBJECTIVE: This study assessed the efficacy and tolerability of diclofenac-K 12.5 mg versus ibuprofen 200 mg and placebo against influenza-like symptoms (oral temperature > or = 38.1 degrees C, and at least moderate headache and muscle/joint aches and pains). METHODS: This was a 3-day, multicenter, randomized, double-blind, double dummy, placebo- and active-controlled, parallel-group trial conducted in Germany. The flexible dosing regimens comprised 2 tablets of diclofenac-K (12.5 mg), ibuprofen (200 mg), or placebo, then 1 to 2 tablets every 4 to 6 hours as needed, to a maximum of 6 tablets/d. Primary efficacy outcomes were fever reduction after the initial dose and an end-of-study global assessment of overall symptom relief. Secondary efficacy outcomes included reduction of feverishness; relief of headache; and relief of muscle/joint aches and pains, after the first dose and at the end of days 1, 2, and 3. Use of rescue medication (paracetamol 500-mg tablets) and usage patterns of study medication were also studied. Tolerability was monitored by recording of patients' adverse events. RESULTS: A total of 356 patients were enrolled (n = 121, 120, and 115 patients in the diclofenac-K, ibuprofen, and placebo groups, respectively). All patients were white except 1 Asian patient in the diclofenac-K group; 55% to 60% of patients in all 3 groups were male; the mean age in each treatment group was approximately 40 years. At baseline, mean oral temperature ranged from 38.65 degrees C in the placebo group to 38.74 degrees C in the diclofenac-K group. Mean oral temperatures in both active groups were significantly lower than that of the placebo group from 30 minutes through 6 hours (P < 0.001), dropping 0.85 degrees C after 4 hours in the diclofenac-K group and 0.76 degrees C in the ibuprofen group versus 0.32 degrees C for placebo. In the end-of-study global treatment assessment, 89.0% of diclofenac-K and 89.1% of ibuprofen patients rated global efficacy as "good" to "excellent" versus only 32.1% for placebo. Diclofenac-K was superior to placebo (P < 0.001) and similar to ibuprofen on all direct assessments of fever and aches and pains. Both active treatments were as well tolerated as was placebo. CONCLUSIONS: In this 3-day study, diclofenac-K 12.5 mg taken in a flexible dosing regimen was more effective than placebo in relieving influenza-like symptoms, with comparable tolerability Efficacy and tolerability of diclofenac-K were similar to those of ibuprofen 200 mg. PMID- 12749507 TI - An open-label, randomized, three-way crossover trial of the effects of coadministration of rosuvastatin and fenofibrate on the pharmacokinetic properties of rosuvastatin and fenofibric acid in healthy male volunteers. AB - BACKGROUND: Rosuvastatin and fenofibrate are lipid-regulating agents with different modes of action. Patients with dyslipidemia who have not achieved treatment targets with monotherapy may benefit from the combination of these agents. OBJECTIVE: The effect of coadministration of rosuvastatin and fenofibrate on the steady-state pharmacokinetics of rosuvastatin and fenofibric acid (the active metabolite of fenofibrate) was assessed in healthy volunteers. METHODS: This was an open-label, randomized, 3-way crossover trial consisting of three 7 day treatment periods. Healthy male volunteers received one of the following treatment regimens in each period: rosuvastatin 10 mg orally once daily; fenofibrate 67 mg orally TID; and rosuvastatin + fenofibrate dosed as above. The steady-state pharmacokinetics of rosuvastatin and fenofibric acid, both as substrate and as interacting drug, were investigated on day 7 of dosing. Treatment effects were assessed by construction of 90% CIs around the ratios of the geometric least-square means for rosuvastatin + fenofibrate/rosuvastatin and rosuvastatin + fenofibrate/fenofibrate for the area under the plasma concentration-time curve (AUC) and maximum plasma concentration (derived from analysis of variance of log-transformed parameters). RESULTS: Fourteen healthy male volunteers participated in the study. When rosuvastatin was coadministered with fenofibrate, there were minor increases in the AUC from 0 to 24 hours and maximum concentration (Cmax) of rosuvastatin: the respective geometric least square means increased by 7% (90% CI, 1.00-1.15) and 21% (90% CI, 1.14-1.28). The pharmacokinetic parameters of fenofibric acid were similar when fenofibrate was dosed alone and with rosuvastatin: the geometric least-square means for fenofibric acid AUC from 0 to 8 hours and Cmax decreased by 4% (90% CI, 0.90 1.02) and 9% (90% CI, 0.84-1.00), respectively. The treatments were well tolerated alone and in combination. CONCLUSION: Coadministration of rosuvastatin and fenofibrate produced minimal changes in rosuvastatin and fenofibric acid exposure. PMID- 12749508 TI - Comparison between repaglinide and glimepiride in patients with type 2 diabetes mellitus: a one-year, randomized, double-blind assessment of metabolic parameters and cardiovascular risk factors. AB - BACKGROUND: Repaglinide and glimepiride are relatively new oral hypoglycemic agents. Few data are available concerning their effects on metabolic parameters other than measures of glycemic control. OBJECTIVES: In addition to assessing the effects of repaglinide and glimepiride on glycemic control in patients with type 2 diabetes mellitus, this study also examined the effects of these agents on 3 metabolic parameters known to be cardiovascular risk factors--lipoprotein(a) (Lp[a]), plasminogen activator inhibitor-1 (PAI-1), and homocysteine (Hcy). METHODS: This randomized, placebo-controlled, double-blind trial was conducted at a single center in Italy. Eligible patients were nonsmokers; had no hypertension or coronary heart disease; were taking no hypolipidemic drugs, diuretics, beta blockers, or thyroxin; and had normal renal function. After an initial 4-week placebo washout period, patients were randomized to receive repaglinide 1 mg/d or glimepiride 1 mg/d. The dose of study drug was optimized over an 8-week titration period, which was followed by a 12-month treatment period. Measures of glycemic control (glycated hemoglobin [HbA1c], fasting plasma glucose [FPG], postprandial plasma glucose [PPG], fasting plasma insulin [FPI], postprandial plasma insulin [PPI]) and the other metabolic parameters of interest were assessed after 6 and 12 months of treatment. RESULTS: One hundred twenty-four patients (63 women, 61 men) completed the study, 62 in each treatment group. There were no significant differences in demographic characteristics between groups. After 6 and 12 months of treatment, FPG levels and HbA1c values were significantly reduced from baseline in both groups (6 months, P < 0.05; 12 months, P < 0.01). After 6 months, PPG levels were significantly decreased only in the repaglinide group (P < 0.05 vs baseline); at 12 months, however, PPG levels were significantly reduced from baseline in both groups (P < 0.01 repaglinide, P < 0.05 glimepiride). No significant changes from baseline in FPI or PPI levels were seen in either group at 6 months, although FPI levels were significantly increased in the repaglinide group at 12 months (P < 0.05). Repaglinide significantly lowered levels of Lp(a), PAI-1, and Hcy after 12 months (all, P < 0.05 vs baseline). Glimepiride significantly lowered levels of Lp(a) and Hcy after 6 months (both, P < 0.05 vs baseline) and levels of Lp(a) (P < 0.01 vs baseline), Hcy (P < 0.01 vs baseline), and PAI-1 (P < 0.05 vs baseline) after 12 months. CONCLUSIONS: Repaglinide and glimepiride improved glycemic control and reduced levels of other metabolic parameters of interest in this population of patients with type 2 diabetes. It is possible that the reductions in Lp(a), PAI-1, and Hcy were the result of improved glucose metabolism; however, the possibility that repaglinide and glimepiride may have a direct effect on these parameters should not be excluded. PMID- 12749509 TI - Levofloxacin compared with imipenem/cilastatin followed by ciprofloxacin in adult patients with nosocomial pneumonia: a multicenter, prospective, randomized, open label study. AB - BACKGROUND: Therapy of nosocomial pneumonia is usually empiric and includes > or = 1 broad-spectrum antimicrobial agent. When considering the use of fluoroquinolones in these difficult-to-treat infections--in which drug delivery to the site of infection may be impaired or organisms with higher minimum inhibitory concentrations may be present--an agent should be chosen whose pharmacodynamics ensure maximal drug exposure. Use of the 750-mg dose of levofloxacin should enhance therapeutic benefit in patients with nosocomial pneumonia. OBJECTIVE: The goal of this study was to compare the efficacy and safety of levofloxacin 750 mg and imipenem/cilastatin followed by ciprofloxacin in adult patients with nosocomial pneumonia. METHODS: This was a multicenter, prospective, randomized, open-label trial conducted in North America. Patients were randomly assigned to 1 of 2 treatment arms: levofloxacin 750 mg QD given i.v. and then orally for 7 to 15 days or imipenem/cilastatin 500 mg to 1 g i.v. every 6 to 8 hours, followed by oral ciprofloxacin 750 mg every 12 hours for 7 to 15 days. Adjunctive antibacterial therapy was mandatory in patients with documented or suspected Pseudomonas aeruginosa or methicillin-resistant Staphylococcus aureus infection. The primary predefined outcome measure was the clinical response (cure, improvement, failure, or unable to evaluate) in microbiologically evaluable patients 3 to 15 days after the end of therapy. RESULTS: The study enrolled 438 adult patients (315 men, 123 women; mean [SD] age, 55.7 [20.04] years). Two hundred twenty patients received levofloxacin, and 218 received the comparator regimen. Demographic and baseline clinical characteristics were similar in the intent-to-treat and clinically evaluable populations. In patients evaluable for microbiologic efficacy, clinical success (cure or improvement) was achieved in 58.1% (54/93) of patients who received levofloxacin, compared with 60.6% (57/94) of patients who received the comparator regimen (95% CI, -12.0 to 17.2). Similar clinical results were seen in patients evaluable for clinical efficacy and in the intent-to-treat population. In the 187 patients evaluable for microbiologic efficacy, eradication was achieved in 66.7% (62/93) of patients receiving levofloxacin and 60.6% (57/94) of patients receiving the comparator regimen (95% CI, -20.3 to 8.3). CONCLUSION: In this study, levofloxacin was at least as effective and was as well tolerated as imipenem/cilastatin followed by ciprofloxacin in adult patients with nosocomial pneumonia, as demonstrated by comparable clinical and microbiologic success rates. PMID- 12749511 TI - Glycemic control in patients with type 2 diabetes mellitus switched from twice daily immediate-release metformin to a once-daily extended-release formulation. AB - BACKGROUND: The extended-release formulation of metformin (MXR) prolongs drug absorption in the upper gastrointestinal tract and permits once-daily dosing in patients with type 2 diabetes mellitus. This newer formulation may enhance patient compliance with oral therapy and improve long-term control of diabetes compared with the conventional immediate-release formulation of metformin (MIR). OBJECTIVE: The goal of this study was to determine the effects on glycemic control of a switch to MXR therapy in patients with type 2 diabetes currently treated with MIR. METHODS: This was a multicenter, randomized, double-blind, parallel-group study in patients with established type 2 diabetes. Eligible patients were to have a glycated hemoglobin (HbA1c) value < or = 8.5% and mean fasting plasma glucose (FPG) concentrations < or = 200 mg/dL while receiving MIR 500 mg BID for at least 8 weeks. After a 2-week, single-blind lead-in period, patients were randomly assigned to receive MXR 1000 or 1500 mg QD for 24 weeks or to continue MIR 500 mg BID for 24 weeks. The primary efficacy variable was change in HbA1c from baseline to week 12. Other variables included change in FPG levels; change in HbA1c; distribution of HbA1c values; mean daily blood glucose concentrations (self-monitored); levels of fructosamine, serum insulin, and lipids; and body weight. RESULTS: Two hundred seventeen patients were randomized to treatment. The mean change from baseline in HbA1c values at weeks 12 and 24 were small and similar in the 3 treatment groups. At week 12, the mean change from baseline in HbA1c was 0.15% for MIR, 0.23% for MXR 1000 mg, and 0.04% for MXR 1500 mg. The corresponding mean changes at week 24 were 0.06%, 0.25%, and 0.14%. CONCLUSIONS: In this study, patients with type 2 diabetes who had been receiving twice-daily MIR achieved comparable glycemic control when therapy was switched to once-daily MXR at the same or a greater total daily dose. PMID- 12749510 TI - Granisetron versus granisetron/dexamethasone combination for the treatment of nausea, retching, and vomiting after major gynecologic surgery: a randomized, double-blind study. AB - BACKGROUND: Granisetron, a selective 5-hydroxytryptamine3 antagonist, is effective for the treatment of patients with postoperative nausea and vomiting. Dexamethasone decreases chemotherapy-induced emesis when added to an antiemetic regimen. OBJECTIVE: This study compared the efficacy of granisetron alone with granisetron/dexamethasone combination for the treatment of nausea and vomiting after major gynecologic surgery. METHODS: In this randomized, double-blind study, patients who were experiencing emetic symptoms during 0 to 3 hours after the end of anesthesia administration received granisetron 40 microg/kg i.v. either alone or in combination with dexamethasone 8 mg. Patients then were observed for 24 hours after study drug administration, with emetic episodes recorded and tolerability assessments performed by nursing staff blinded to treatment. RESULTS: A total of 120 women were enrolled (n = 60 in each treatment group; mean [SD] age in the granisetron group, 44 [9] years [range, 23-63 years]; combination group, 45 [8] years [range, 21-65 years]). No significant differences in patient demographic characteristics were found between the 2 treatment groups. However, the percentage of patients free of emetic symptoms (nausea, retching, vomiting) was higher in the granisetron/dexamethasone combination group than in the granisetron group (95.0% and 80.0%, respectively; P = 0.012). No clinically serious adverse events attributed to the study drugs were observed in either group. CONCLUSION: In this study, the granisetron/dexamethasone combination was more effective than was granisetron alone for the management of nausea and vomiting during 0 to 3 hours after anesthesia in women undergoing major gynecologic surgery. PMID- 12749512 TI - A post hoc analysis of the impact on hostility and agitation of quetiapine and haloperidol among patients with schizophrenia. AB - BACKGROUND: Quetiapine, a drug with a broad pharmacologic profile (similar to that of clozapine), may show benefits for agitation in patients with psychoses. Also, quetiapine may be superior to placebo and either equal or superior to haloperidol in treating this symptom. Available data for other second-generation antipsychotic agents show that quetiapine may have better efficacy in improving agitation compared with haloperidol. OBJECTIVE: This reanalysis of a previously reported pivotal clinical trial assessed whether quetiapine or haloperidol has benefits for the treatment of hostility and agitation among patients experiencing an acute exacerbation of schizophrenia. METHODS: Patients aged 18 to 65 years of either sex and any ethnicity who had a diagnosis of schizophrenia based on the Diagnostic and Statistical Manual of Mental Disorders, Revised Third Edition criteria and who were experiencing an acute exacerbation were recruited into the study. A priori, data from patients assigned to 4 therapeutically effective quetiapine treatment groups (150, 300, 600, and 750 mg) in a previously reported 6-week, double-blind, placebo-controlled clinical trial were combined and compared with data from patients given haloperidol 12 mg or placebo on an agitation measure derived from the Brief Psychiatric Rating Scale (BPRS). Patients who received at least 2 weeks of treatment who had a baseline assessment and at least 1 postbaseline assessment after the 2 weeks of treatment were included. An analysis of variance with the baseline hostility score and center as covariates was used to assess treatment effects of quetiapine or haloperidol versus placebo for changes in agitation scores. A path analysis was used to separate the direct from the indirect effects (via improvements in psychoses and/or overall psychopathology) on agitation scores of quetiapine relative to haloperidol. RESULTS: A total of 257 patients (193 men, 64 women) were studied. The combined quetiapine groups comprised 175 patients; the haloperidol group, 42 patients; and the placebo group, 40 patients. Quetiapine treatment reduced agitation scores significantly among patients with acute psychoses compared with placebo. A slight reduction in agitation scores was found when haloperidol treatment was compared with placebo, but this difference was not statistically significant. Compared with haloperidol, quetiapine treatment had a direct and significant effect on agitation that was independent of the improvement in psychotic symptoms. CONCLUSIONS: The data in this study suggest that quetiapine treatment has benefits for hostility and agitation among patients experiencing an acute exacerbation of schizophrenia. Furthermore, the path analysis indicated that, relative to haloperidol, quetiapine appeared to have direct effects on agitation that were independent of improvements in psychoses or overall psychopathology, as assessed by the BPRS. PMID- 12749513 TI - A comparison of 5-day courses of dirithromycin and azithromycin in the treatment of acute exacerbations of chronic obstructive pulmonary disease. AB - BACKGROUND: Short-term use of antibiotics has become a common component of the management of acute exacerbations of chronic bronchitis (AECB), particularly in complex cases with productive cough or purulent phlegm. The macrolide antibiotics, particularly second-generation agents such as dirithromycin and azithromycin, are among the antibiotic classes frequently recommended and used to treat upper and lower respiratory infections, including AECB. OBJECTIVE: This study compared the clinical efficacy and tolerability of 5-day courses of dirithromycin and azithromycin given once daily for the treatment of acute exacerbations of chronic obstructive pulmonary disease (COPD). METHODS: This randomized, investigator-blinded, parallel-group clinical trial was conducted at 5 centers in the United States. Eligible patients were adult (age >35 years) smokers or ex-smokers (smoking history of at least 10 pack-years) with chronic bronchitis and an acute exacerbation, defined by the occurrence of increased dyspnea and/or productive cough and feverishness within 48 hours of enrollment. Before randomization, an attempt was made to obtain a sputum specimen from each patient for Gram's staining and culture. Patients were randomized to receive dirithromycin 500 mg QD for 5 days or azithromycin 500 mg QD on day 1 and 250 mg QD on days 2 to 5. Clinical efficacy was assessed separately by patients and physicians at early (days 7-10) and late (days 25-35) posttreatment visits. RESULTS: Eighty-six patients (48 women, 38 men; mean age, 55 years) with a mean smoking history of 31 pack-years were included in the intent-to-treat analysis. Forty-six (54%) patients were randomized to dirithromycin and 40 (47%) patients to azithromycin. Clinical efficacy was reported in a high proportion of patients in both treatment groups, both at the early posttreatment visit (84.8% dirithromycin, 75.7% azithromycin; difference dirithromycin - azithromycin, 9.1%; 95% CI, -8.2 to 26.4) and the late posttreatment visit (95.5% and 86.5%, respectively; difference dirithromycin - azithromycin, 9.0%; 95% CI, -3.7 to 21.6). A similar proportion of patients required a second course of antibiotics over the study period (20.5% dirithromycin, 27.0% azithromycin; difference dirithromycin - azithromycin, -6.6%; 95% CI, -25.2 to 12.1). Only 42 (48.8%) patients were able to produce a sputum sample before receiving study treatment, and of these, only 20 (47.6%) demonstrated a preponderance of neutrophils on Gram's staining. Both treatments were well tolerated. CONCLUSIONS: The results of this study suggest comparable clinical efficacy between 5-day courses of once daily dirithromycin and azithromycin in acute exacerbations of COPD. There were insufficient data to permit meaningful comparison of the bacteriologic efficacy of these macrolide antibiotics. PMID- 12749514 TI - Pilot study of a survey of US residents purchasing medications in Mexico: demographics, reasons, and types of medications purchased. AB - BACKGROUND: Medications sold only by prescription in the United States are easily obtained in Mexican pharmacies without prescriptions. Few studies have researched the impact of this phenomenon in the United States. OBJECTIVE: The aim of this pilot study was to determine the demographics of Americans traveling to Mexico to purchase medications, their reasons for doing so, and which medications they purchased. METHODS: US residents purchasing medications in Nuevo Progreso, Mexico, were given a voluntary, self-administered questionnaire. Some questions had more than 1 potential answer. RESULTS: A total of 103 pharmacy consumers were approached, of whom 100 agreed to participate; no data were gathered on the 3 pharmacy consumers who declined to participate. The survey results indicated that 60.0% of participants (60 participants) had private insurance, 18.0% (18 participants) had Medicare, 3.0% (3 participants) had some other form of insurance, and 19.0% (19 participants) had no insurance at all. The primary reason cited by participants for traveling to Mexico to purchase medications was lower prices (accounting for 66.7% of all reasons given). Infection was the primary disease treated, accounting for 28.2% of types of conditions cited (42/149), and ampicillin was the drug most frequently purchased, accounting for 16.8% (25/149) of all medication purchases. CONCLUSIONS: This pilot study indicates that US residents travel to Mexico to purchase medications for many reasons and for many different medical conditions, including chronic conditions. The most frequently cited reason for purchasing medications in Mexico was lower prices, even among participants with some form of health insurance. This finding suggests that until drug pricing is further addressed in the United States, residents will continue to seek lower-cost alternatives for purchasing their medications, including travel to Mexico. PMID- 12749515 TI - Comparison of peptic-ulcer drug use and expenditures before and after the implementation of a government policy to separate prescribing and dispensing practices in South Korea. AB - BACKGROUND: The South Korean government instituted a new policy, the separation of prescribing and dispensing (SPD) of medications, on July 1, 2000, to provide greater differentiation between the roles of physicians and pharmacists than had historically existed in South Korea. It was hoped that this policy would promote the rational use of medications and reduce medication expenditures, which accounted for approximately 30% of the total health care expenditures before the implementation of SPD. OBJECTIVE: The purpose of this study was to assess the effects of SPD on drug market share and expenditures for branded and generic medications by comparing the use of and expenditures for peptic-ulcer medications before and after the implementation of SPD. METHODS: Data on expenditures and quantity of use in January and December 2000 (in terms of defined daily dose [DDD]) of peptic-ulcer medications were obtained from the Korean National Health Insurance claims database. These data were derived using a 3-stage probability sample of prescription data from medical clinics in South Korea. RESULTS: The number of prescription drug claims for peptic-ulcer drugs increased by 13.9% after the introduction of SPD. Medication expenditures increased by 98.4% for peptic-ulcer medications. The use of more expensive drugs and branded products, even when generic products were available, accounted for most of this increase. In particular, the use of branded ranitidine 150 mg (measured by DDD) increased from 6.3% of the market share before SPD to 27.6% of the market share after the implementation of SPD. CONCLUSIONS: The implementation of SPD increased both prescription drug claims and expenditures for peptic-ulcer medications. A principal factor contributing to the increase in expenditures was the use of branded medications. PMID- 12749516 TI - Factors associated with hospitalization costs for patients with community acquired pneumonia. AB - BACKGROUND: Combined mortality rates for pneumonia and influenza suggest that the 2 conditions represent the sixth leading cause of death in the United States. The total cost of pneumonia, including indirect costs, was estimated to be approximately $23 billion per year in 1994. OBJECTIVE: The objective of this study was to assess variables that may be significantly associated with the cost of treating patients hospitalized with community-acquired pneumonia (CAP). We also assessed the impact of treatment guidelines for management of CAP (developed by managed care plans) on total costs. METHODS: Patients in 3 managed care plans who were hospitalized with a primary or secondary diagnosis of CAP in Maryland and Washington, DC, between January 1, 1997, and April 30, 1997, or between January 1, 1998, and April 30, 1998, were identified based on International Classification of Diseases, Ninth Revision codes. Clinical data were abstracted from patients' medical charts by nurses, and billing data were acquired from these plans. A retrospective data analysis was carried out using billing data from 3 managed care plans and clinical data from hospitals associated with the plans. A multivariate regression model was developed using the natural logarithm of cost as the dependent variable. Independent variables that were studied included severity of illness, days in the intensive care unit (ICU), triage per guidelines, drug therapy per guidelines, mortality, and managed care plan identifiers. RESULTS: The charts of 569 patients were assessed. The mean age of the study sample was 75.3 years. ICU days (P < 0.001), mortality, and drug therapy (both P < 0.01) per guidelines significantly affected costs. As expected, an increase in the number of ICU days led to an increase in costs. However, patients who received drug therapy recommended by the guidelines had significantly lower costs than patients not treated according to the guidelines (P = 0.001). CONCLUSIONS: The findings of this study suggest that guidelines for CAP management, such as those developed by managed care plans, may help reduce costs by minimizing unnecessary ICU admissions and appropriately managing patients with CAP. PMID- 12749517 TI - A cost-utility analysis of mitoxantrone hydrochloride and interferon beta-1b in the treatment of patients with secondary progressive or progressive relapsing multiple sclerosis. AB - BACKGROUND: Information on the cost utility of interferon beta-1b and mitoxantrone hydrochloride, 2 disease-modifying agents approved by the US Food and Drug Administration for the treatment of secondary progressive (SP) multiple sclerosis (MS), is limited. OBJECTIVE: The aim of this study was to compare the cost utility of i.v. mitoxantrone hydrochloride administered every 3 months, s.c. interferon beta-1b administered every other day, and routine supportive care from the perspectives of both the insurer and society. METHODS: We used a Markov model with health states based on the Kurtzke Expanded Disability Status Scale (EDSS) scores from both an insurer's and a societal perspective (including direct and total costs, respectively). Theoretical patients entered the model with an EDSS score of 3; their progression was followed for 10 years. Transition probabilities were derived from clinical trial data. Cost and utility inputs were taken from the literature. Sensitivity analyses were conducted on all variables. RESULTS: From the insurer's perspective, the incremental cost-utility ratio of mitoxantrone hydrochloride therapy compared with routine supportive care was 58,272 dollars per quality-adjusted life year (QALY) gained. From a societal perspective, mitoxantrone hydrochloride was more effective and less costly than supportive care. From the perspectives of insurers and society, the cost-utility ratios of interferon beta-1b compared with routine supportive care were 338,738 dollars and 245,700 dollars per QALY gained, respectively. When compared with mitoxantrone hydrochloride, interferon beta-1b had an incremental cost-utility ratio of 741,331 dollars and 658,402 dollars per QALY from the insurer's and society's perspectives, respectively. Cost-utility ratios for mitoxantrone hydrochloride were sensitive to acquisition and administration costs of therapy and to effectiveness at slowing disease progression. Cost-utility ratios for interferon beta-1b were not sensitive to any of the variables included in the model. CONCLUSIONS: Mitoxantrone hydrochloride is likely to be a cost-effective treatment for patients with SPMS or progressive relapsing MS from an insurer' perspective and is cost saving from a societal perspective. Interferon beta-1b is not likely an efficient treatment using conventional comparisons for cost effectiveness. This analysis has potentially important implications for policy implementation; however, decisions about which agent to use for each patient should consider the treatment's adverse-event profile, the method of administration, and the patient's preferences for these factors. PMID- 12749518 TI - A pharmacoeconomic comparison of the efficacy and costs of pantoprazole and omeprazole for the treatment of peptic ulcer or gastroesophageal reflux disease in The Netherlands. AB - BACKGROUND: The focus of treatment of patients with peptic ulcer or gastroesuphageal reflux disease has changed during the last 15 years, with a shift from histamine2-receptor antagonists to proton-pump inhibitors (PPIs). From 1993 to 2000, expenditures for omeprazole (90% of total market share of PPIs) increased in The Netherlands from 68 million euros to 230 million euros. In 1999, expenditures for pantoprazole accounted for the majority of the rest of the market share for PPIs. OBJECTIVE: The objective of this study was to compare the efficacy and costs of treatment with pantoprazole and omeprazole in The Netherlands. METHODS: First, we reviewed clinical studies that compared the efficacy of different dosages of omeprazole and pantoprazole. Second, we analyzed data from a nationwide database of drug prescriptions to determine the dosages used in daily practice in 1999. The data were based on a representative sample of approximately 40% of the Dutch community pharmacies. Third, we modeled the outcome of potential substitution of pantoprazole for omeprazole and the corresponding scenarios for nationwide cost savings using the prescription information from the nationwide database. Potential savings within the Dutch health care system were estimated. RESULTS: The 1999 prescription data indicated that pantoprazole treatment cost a mean 1.59 euros/d, compared with 2.12 euros/d for omeprazole (1.00 euro = 1.0487 US dollars). The mean cost per defined daily dose of omeprazole was 1.65 euros, compared with 1.59 euros for pantoprazole. Following the summary of product characteristics, treatment with pantoprazole appeared to be less costly for all indications. The projected annual cost savings for substituting pantoprazole for omeprazole on 90% of treatment days were estimated at 40.8 million euros. However, these projected savings may be offset by the costs of switching and the costs of upward dose adjustments that some patients may require. CONCLUSIONS: Based on the available documentation about effectiveness and costs of omeprazole and pantoprazole, pantoprazole may provide a more favorable pharmacoeconomic profile than omeprazole. However, this is only true if the substitution of omeprazole by pantoprazole can be achieved without loss of efficacy or tolerability. PMID- 12749519 TI - A model analysis of costs of blood pressure destabilization and edema associated with rofecoxib and celecoxib among older patients with osteoarthritis and hypertension in a Medicare Choice population. AB - BACKGROUND: Economic analyses consider all costs relevant to the use of a particular treatment or treatments. Recently, head-to-head, randomized, controlled trials have shown a significantly higher incidence of blood pressure (BP) destabilization and clinically significant edema with rofecoxib than with celecoxib among older, hypertensive patients with osteoarthritis (OA). OBJECTIVE: The objective of this analysis was to estimate the COX-2 specific inhibitor medication costs, in addition to the costs of drugs and physicians' fees, for BP destabilization and clinically significant edema associated with the use of rofecoxib 25 mg QD and celecoxib 200 mg QD in patients with OA and hypertension in a Medicare Choice population (aged > or = 65 years). METHODS: A decision analysis model was constructed to determine the costs (from the payer's perspective) of treating patients in this population with either of the 2 regimens for 6 weeks. The analysis used pooled data from 2 recent, independently conducted, multicenter, double-blind, randomized, controlled trials of OA patients aged > or = 65 years with treated hypertension who received either celecoxib 200 mg QD or rofecoxib 25 mg QD for 6 weeks. In the individual trials, rofecoxib was associated with significantly higher rates of destabilized BP (P < 0.032 and P < 0.001) and edema (P < 0.01 and P = 0.045) than celecoxib. RESULTS: For a 100,000-member Medicare Choice population, an estimated 25,630 persons would have OA and hypertension (stages I-III), and an estimated 5126 of these patients would use celecoxib or rofecoxib. The estimated costs were 33,938 dollars (6.2%) higher if all hypertensive patients with OA were treated with rofecoxib rather than celecoxib for 6 weeks. The cost per day of use was 0.16 dollars less with celecoxib, and per-patient, per-month costs were 4.79 dollars lower. CONCLUSION: Celecoxib was a less costly treatment option than rofecoxib among OA patients with hypertension aged > or = 65 years, based on our model of the direct costs of COX-2 specific inhibitor therapy combined with those associated with physician monitoring and treatment of edema and BP destabilization. PMID- 12749520 TI - Outcomes research in oncology: improving patients' experiences with cancer treatment. AB - Outcomes research in oncology is a relatively young field, but its potential for expanding our understanding of patients' experiences with cancer gives it increasing relevance to clinical oncology research. We provide a brief overview of the growing prevalence of oncology outcomes research, and we discuss some of the key areas of inquiry currently engaging outcomes researchers. In doing so, we introduce the articles in this supplemental section, which address some of the unique concerns of outcomes researchers and outline the most important challenges confronting this research community. PMID- 12749521 TI - Outcomes research related to patient decision making in oncology. AB - For outcomes research, what are the implications of seeing the patient as a decision maker? In the current medical environment, greater emphasis is placed on the role played by the patient in clinical decision making. In the past 2 decades, considerable work has been done to identify and measure decision-related outcomes, including knowledge about the treatment options (risks and benefits), satisfaction, anxiety, decisional conflict, and involvement in the decision process. Attempts to improve these decision-related outcomes involve patient decision aids, which convey patient-specific information and sometimes help patients proceed through an explicit decision-making process. These interventions have produced positive results, especially with respect to improving patient knowledge. Future research is needed to understand which aspects of the interventions work and for what types of patients. Research is also needed to better understand the decision making process of patients who do not use decision aids. PMID- 12749523 TI - Outcomes research at the National Cancer Institute: measuring, understanding, and improving the outcomes of cancer care. AB - What is the National Cancer Institute (NCI) doing to enhance the state of the science for measuring and understanding patient-centered outcomes of cancer care and to make this information useful for improved decision making? The NCI has a new focus on research that describes, interprets, and predicts the impact of various influences, especially interventions, on end points that matter to decision makers. The research includes end points such as survival, health related quality of life, satisfaction and patient experience, and economic burden. To further this work, NCI supports and conducts research to (1) identify valid, reliable, responsive, and feasible end-point measures; (2) collect high quality evidence about the impact of interventions on the end points of interest; (3) improve our understanding of the effects of other factors that shape this interaction; and (4) expand our capacity to translate research findings into information that is useful to patients, clinical policy makers, payers, regulators, standard setters, and providers of cancer care. PMID- 12749522 TI - Trends in clinical trials in surgical oncology: implications for outcomes research. AB - Outcomes analysis is emerging as a crucial tool in understanding and improving health care. To this end, outcomes research is focused on methodologies for assessing the impact and quality of clinical services and the penetration of new findings. Clinical trials are an important mechanism for generating data to support outcomes research activities. In clinical trials in surgical oncology, there has been a general broadening of the spectrum of end points that are more focused on the needs of providing data to outcomes researchers. For example, in addition to traditional "mechanical" end points reflecting the physical dimensions of disability, awareness of the importance of end points related to quality of life has increased. Moreover, a variety of innovative study designs and methodologies have found more widespread use, including noninferiority trials, preresection staging designed to direct therapy more precisely, planned early reports of conditional results, prognostic studies, assessment of an event status at a point in time, and large simple trials. The goal of this article is to summarize and assess some of the trends in clinical trials in surgical oncology. Examples of ongoing studies or of trials in development from the American College of Surgeons Oncology Group are discussed. PMID- 12749524 TI - A randomized trial of telenursing to reduce hospitalization for heart failure: patient-centered outcomes and nursing indicators. AB - This trial compared 3 post-hospitalization nursing care models for reducing congestive heart failure (CHF) readmission charges during 180-days of follow-up. Subjects received in-person visits at baseline and 60 days plus 1 of 3 care modalities in the interim: (a) video-based home telecare; (b) telephone calls; and (c) usual care. CHF-related readmission charges were > 80% lower in the telenursing groups compared to usual care, and these groups also had significantly fewer CHF-related emergency visits. In-person visits were more than 3 times longer than telenursing visits (p < 0.0001), only partially due to added travel time. Patient self-care adherence, medications, health status, and satisfaction did not significantly differ between groups. Telenursing can reduce CHF hospitalizations and allow increased frequency of communication with patients. PMID- 12749525 TI - A comparative analysis of disability measures and their relation to home health care use. AB - This study examines the relationship between various definitions of disability and the use of home health care. Using data from the 1993 Survey of Asset and Health Dynamics among the Oldest Old, four measures of disability are constructed based on the concepts of difficulty and assistance. Logistic regression techniques are used to examine differences across measures in the probability of home health care use for specific subgroups of the older population (e.g., married Black females, unmarried White males). Home health care use was most likely when the presence of both assistance and difficulty were included in the definition of disability. Regardless of the measure used, women, Whites, and unmarried people had higher likelihoods of using home health care than men, Blacks, and married people respectively. If used as a service eligibility criterion, an inclusive measure of disability would result in a large number of potential service users. Efforts to control expenditures through the use of a narrower measure that requires the presence of assistance may disadvantage some subgroups of the older population. PMID- 12749526 TI - Frequency and determinants of urgent requests to home care agencies for community dwelling elderly. AB - We analyzed a one-year case series and performed a longitudinal (4 month) cohort analysis of urgent requests made to home care agencies by and for their > or = 65 years old clients in order to estimate the frequency of unscheduled services delivered by home care agencies and to identify risk factors. All 40 home care agencies located in a Swiss region were included in the study and we registered 3,816 urgent requests (75/1,000 > or = 65 years residents per year). Among home care users, the presence of a urinary catheter, incontinence and the need for assistance in bathing were predictors of unscheduled services. Resources should be planned in order to help home care teams to handle unexpected, disruptive clusters of urgent requests that may compromise their scheduled activities. PMID- 12749527 TI - Evolving the delivery of acute care services in the home. AB - To serve escalating acute care caseloads, physicians affiliated with one Canadian home care program have piloted a project to integrate physician services into the home (IPSITH). This paper presents the 18-month qualitative evaluation. Phenomenological methodology and in-depth interviewing were used to construct a holistic interpretation of the implementation from the experiences of all involved: patients, family caregivers, physicians, case managers, community nurses and the project's nurse practitioner. Findings revealed the central role of the nurse practitioner, who served as a clinical expert, care coordinator and case manager. Several unsolved issues were identified: the extent to which home care is a viable alternative to hospitalization, the feasibility of physician involvement, redundancies with hospital emergency services, and the limitations of system resources for funding such services. The researchers conclude that full scale long-term integration of physician services in the home may require macro level decisions about system design, resource allocation, and professional regulations. PMID- 12749528 TI - Acute health care service use among elderly home care clients. AB - Utilization of acute health care services accounts for a substantial proportion of health expenditures in Canada, and is associated with compromised health and autonomy for older persons. Using the Resident Assessment Instrument for Home Care (RAI-HC), this cross-sectional study of 683 elderly home care recipients sought to distinguish clients who were more likely to use acute health care services; i.e., hospital admissions, emergency room visits. Clients with nutritional problems were 2.58 times more likely to have used acute health care services than clients without nutritional problems. Among clients with a poor social support system, those with nutrition problems were 5.95 times as likely to have used acute health care services. Poor self-rated health, and greater functional dependency were also signif- icantly associated with acute health care use. This study provides a profile of elderly home care clients who are at risk of using acute health care services, which may facilitate targeted efforts to prevent unplanned acute health care use. PMID- 12749529 TI - Clonal exhaustion as a result of immune deviation. AB - An overwhelming virus infection that spreads within a few days throughout the host can cause deletion of the specific cytotoxic T lymphocytes (CTL). This phenomenon is known as 'clonal exhaustion'. Current explanations for this phenomenon are 'clonal', and consider either the terminal differentiation of the virus-specific CTL to an effector phenotype, or the lack of help and antigen presentation for a specific CTL clone. The virus remains controlled by some other form of immunity in the exhausted state. Candidates are innate immunity (especially NK cells and macrophages) and a T helper type 2 based immune response. Surprisingly, the role of this other form of immunity in causing exhaustion has been ignored so far. Developing a mathematical model, we here investigate the possibility that this inter-clonal immunity is responsible for exhaustion by down regulating the CTL response. The model is based on previously published exhaustion data for Lymphocytic choriomeningitis virus as an in vivo model. We demonstrate that several complicated experiments on clonal exhaustion are consistent with inter-clonal regulation. By interpreting the available data with a mathematical model, we compare this novel mechanism with the mechanisms suggested previously. PMID- 12749530 TI - Proliferation and competition in discrete biological systems. AB - We study the emergence of collective spatio-temporal objects in biological systems by representing individually the elementary interactions between their microscopic components. We use the immune system as a prototype for such interactions. The results of this detailed explicit analysis are compared with the traditional procedure of representing the collective dynamics in terms of densities that obey partial differential equations. The simulations show even for very simple elementary reactions the spontaneous emergence of localized complex structures, from microscopic noise. In turn the effective dynamics of these structures affects the average behaviour of the system in a very decisive way: systems which would according to the differential equations approximation die, display in reality a very lively behaviour. As the optimal modelling method we propose a mixture of microscopic simulation systems describing each reaction separately, and continuous methods describing the average behaviour of the agents. PMID- 12749531 TI - The duration of a sequence of epidemics. AB - The typical duration of an epidemic in a sequence of linearly ordered populations shows a surprising nonmonotonic behaviour with respect to population size, which was noted by Swinton (1998) [Bull. Math. Biol., 60, 215-230]. This paper gives the sketch of a proof of the phenomenon. PMID- 12749532 TI - Optimal scheduling of radiotherapy and angiogenic inhibitors. AB - We incorporate a previously validated mathematical model of a vascularized tumor into an optimal control problem to determine the temporal scheduling of radiotherapy and angiogenic inhibitors that maximizes the control of a primary tumor. Our results reveal that optimal antiangiogenic monotherapy gives a large initial injection to attain a 20:1 ratio of tumor cell volume to supporting vasculature volume. It thereafter maintains this 20:1 ratio via a continuous dose rate that is intensified over time. The optimal radiation monotherapy schedule is characterized by a modest dose intensification over time. The best performance is achieved by our optimal combination regimen, where the antiangiogenic treatment again maintains a constant tumor-to-vasculature ratio, but is administered in a dose-intensified manner only during the latter portion of the radiation fractionation schedule. PMID- 12749533 TI - Quantification of the spatial aspect of chaotic dynamics in biological and chemical systems. AB - The need to study spatio-temporal chaos in a spatially extended dynamical system which exhibits not only irregular, initial-value sensitive temporal behavior but also the formation of irregular spatial patterns, has increasingly been recognized in biological science. While the temporal aspect of chaotic dynamics is usually characterized by the dominant Lyapunov exponent, the spatial aspect can be quantified by the correlation length. In this paper, using the diffusion reaction model of population dynamics and considering the conditions of the system stability with respect to small heterogeneous perturbations, we derive an analytical formula for an 'intrinsic length' which appears to be in a very good agreement with the value of the correlation length of the system. Using this formula and numerical simulations, we analyze the dependence of the correlation length on the system parameters. We show that our findings may lead to a new understanding of some well-known experimental and field data as well as affect the choice of an adequate model of chaotic dynamics in biological and chemical systems. PMID- 12749534 TI - Growth and function of fungal mycelia in heterogeneous environments. AB - As decomposer organisms, pathogens, plant symbionts and nutrient cyclers, fungi are of fundamental importance in the terrestrial environment. Moreover, in addition to their well-known applications in industry, many species also have great potential in environmental biotechnology. The study of this important class of organisms is difficult through experimental means alone due to the heterogeneity of their natural growth habitat and the microscopic scale of growth. In this work we present a mathematical model for colony expansion that is derived through consideration of the growth characteristics on the microscale. The model equations are of mixed hyperbolic-parabolic type and are treated with a numerical scheme that preserves positivity and conserves mass. The numerical solutions are compared against experimental results in a variety of environments. Thus the effect of different translocation mechanisms on fungal growth and function are identified. The derivation and analysis of an approximation to the full model yields further results concerning basic properties of mycelial growth. Finally, the acidification of the growth habitat is considered and the model thus provides important predictions on the functional consequences of the redistribution of internally-located material. PMID- 12749535 TI - Multiple attractors in stage-structured population models with birth pulses. AB - In most models of population dynamics, increases in population due to birth are assumed to be time-independent, but many species reproduce only during a single period of the year. A single species stage-structured model with density dependent maturation rate and birth pulse is formulated. Using the discrete dynamical system determined by its Poincare map, we report a detailed study of the various dynamics, including (a) existence and stability of nonnegative equilibria, (b) nonunique dynamics, meaning that several attractors coexist, (c) basins of attraction (defined as the set of the initial conditions leading to a certain type of attractor), (d) supertransients, and (e) chaotic attractors. The occurrence of these complex dynamic behaviour is related to the fact that minor changes in parameter or initial values can strikingly change the dynamic behaviours of system. Further, it is shown that periodic birth pulse, in effect, provides a natural period or cyclicity that allows multiple oscillatory solutions in the continuous dynamical systems. PMID- 12749536 TI - Biodiversity, habitat area, resource growth rate and interference competition. AB - For the majority of species, per capita growth rate correlates negatively with population density. Although the popular logistic equation for the growth of a single species incorporates this intraspecific competition, multi-trophic models often ignore self-limitation of the consumers. Instead, these models often assume that the predator-prey interactions are purely exploitative, employing simple Lotka-Volterra forms in which consumer species lack intraspecific competition terms. Here we show that intraspecific interference competition can account for the stable coexistence of many consumer species on a single resource in a homogeneous environment. In addition, our work suggests a potential mechanism for field observations demonstrating that habitat area and resource productivity strongly positively correlate to biodiversity. In the special case of a modified Lotka-Volterra model describing multiple predators competing for a single resource, we present an ordering procedure that determines the deterministic fate of each specific consumer. Moreover, we find that the growth rate of a resource species is proportional to the maximum number of consumer species that resource can support. In the limiting case, when the resource growth rate is infinite, a model with intraspecific interference reduces to the conventional Lotka-Volterra competition model where there can be an unlimited number of coexisting consumers. This highlights the crucial role that resource growth rates may play in promoting coexistence of consumer species. PMID- 12749537 TI - Traveling waves of infection in the hantavirus epidemics. AB - Traveling waves are analyzed in a model of the hantavirus infection in deer mice. The existence of two kinds of wave phenomena is predicted. An environmental parameter governs a transition between two regimes of propagation. In one of them the front of infection lags behind at a constant rate. In the other, fronts of susceptible and infected mice travel at the same speed, separated by a constant delay. The dependence of the delay on system parameters is analyzed numerically and through a piecewise linearization. PMID- 12749538 TI - Dynamical analysis of a degenerate primary and secondary humoral immune response. AB - Lymphocyte receptor response to antigen is degenerate. Each receptor can have a high affinity to more than one antigen. The optimal level of degeneracy was previously modeled using different methods; all showing that the degeneracy level should be inversely proportional to the probability that an antigen belongs to the self repertoire. Here we develop a new formalism, reproducing the results of previous models, which enables us to study the relation between receptor degeneracy and the pathogen-immune cell interaction dynamics, in primary and secondary response. We begin by developing a general formalism and reproducing the results obtained by Nemazee: (1) that an optimal immune system will have a capacity which is inversely proportional to the fraction of self-antigens and (2) that the number of self-reactive cells that the body destroys is tuned by this capacity optimization to be 63%. We then use our extended framework to relate the minimal number of B cell precursor required to mount an immune response to the naive B cell production rate. Finally, we analyze the dynamics of the interaction between the immune system and a pathogen and show that memory cells may be used as the first line of defense, while newly created cells are used later to refine the immune response. PMID- 12749539 TI - Quantifying consequences of chronic sleep restriction. PMID- 12749540 TI - In pursuit of knowledge. PMID- 12749541 TI - Competency-based goals for sleep and chronobiology in undergraduate medical education. PMID- 12749542 TI - 2003 National Sleep Disorders Research Plan. PMID- 12749543 TI - Long-lasting enhancement of rapid eye movement sleep and pontogeniculooccipital waves by vasoactive intestinal peptide microinjection into the amygdala temporal lobe. AB - STUDY OBJECTIVES: The effect of a vasoactive intestinal peptide (VIP) microinjection into the amygdaloid central (CN) and basal nuclei (BN) on sleep organization and on the number and pattern of occurrence of pontogeniculooccipital (PGO) waves was analyzed. DESIGN: One group of 8 cats was studied in baseline conditions and after the microinjection of two doses of VIP applied into the CN and BN. SETTING: Sleep research laboratory. PARTCIPANTS AND INTERVENTIONS: Eight cats were prepared with sleep-recording electrodes and with guide tubes in both amygdalae for saline and VIP microinjections. Neuropeptide doses of 0.10 microg/1 microl (30 microM) and 0.33 microg/1 microl (99.24 microM) were employed. MEASUREMENTS AND RESULTS: Once the microinjection was applied, 23 hour polygraphic sleep recordings were performed for 5 consecutive days. Concomitantly the PGO waves were tape-recorded on each day and computationally analyzed. Results show that the 0.10 microg/1 microl microinjection produced no change. Unilateral VIP 0.33 microg/1 microl injection into the CN provoked a significant and lasting increase in the percentage of slow-wave sleep with PGO waves. Bilateral application of VIP increased the percentage of slow-wave sleep with PGO waves and rapid eye movement sleep for 5 days. Bilateral microinjection of the neuropeptide into the BN only enhanced the percentage of slow wave sleep with PGO waves. For both amygdaloid nuclei, we observed that VIP increased the number and modified the PGO wave pattern of occurrence during slow-wave sleep with PGO waves and during rapid eye movement sleep. CONCLUSIONS: The VIP microinjection into both the CN and BN induces increased amounts of rapid eye movement sleep, PGO waves, and slow-wave sleep with PGO waves, having a more robust effect on all of these three variables when applied into the CN. PMID- 12749544 TI - Sleep in female mice: a strain comparison across the estrous cycle. AB - STUDY OBJECTIVE: Studying inbred strains of mice has proven useful in uncovering genetic variation in the expression of sleep patterns. However, although genetic influence on many behaviors has been shown to be gender specific, to date, sleep patterns in different strains of female mice have not been reported. In order to perform such studies in female mice, the estrous cycle must be taken into account in view of the effects of reproductive hormones on sleep. The aim of this study was thus to determine sleep patterns in female mice of different inbred strains over the estrous cycle. DESIGN: Three strains of mice were used. Vaginal smears were performed to determine the estrous cycle stage; electroencephalographic and electromyographic activity, as well as body temperature and locomotor activity collected during a full estrous cycle, were analyzed. MEASUREMENTS AND RESULTS: We report a major impact of the genetic background in the regulation of non-rapid eye movement sleep over a 24-hour period and clear strain differences in rapid eye movement sleep distribution over the light-dark cycle. In contrast, the estrous cycle had less influence on non-rapid eye movement sleep and rapid eye movement sleep, and these effects were dependent on the genotype of the mice. CONCLUSIONS: Sleep regulation in female mice is influenced primarily by genetic background and, to a lesser extent, by hormonal variations associated with the estrous cycle. PMID- 12749546 TI - Narcolepsy-like symptoms in a patient with down syndrome and without obstructive sleep apnea. AB - OBJECTIVE: The study reports on a 33-year-old Caucasian female with Down syndrome and narcolepsy-like symptoms. METHOD: After medical and genetic screening, docturnal polysomnography followed by a Multiple Sleep Latency Test and HLA typing were performed. The patient was medication free and reported a number of cataplexy attacks everyday. Each time that she came to the sleep disorders clinic, she was observed to have cataplexy. She also felt extremely drowsy. A mean sleep latency of 8.8 minutes with 4 sleep-onset rapid eye movement periods in the Multiple Sleep Latency Test, with no other sleep disorder that accounts for the symptoms, was obtained. The patient was DQB1*0301, DQB1*0602, as revealed by the last high-resolution serologic typing. PMID- 12749545 TI - Fatigue, sleep restriction, and performance in automobile drivers: a controlled study in a natural environment. AB - OBJECTIVES: To test the neurobehavioral consequences of sleep restriction combined with fatigue from long-distance driving (1000 Km/600 miles). DESIGN: Counterbalanced study involving 3 experimental conditions: laboratory after controlled habitual sleep (8.5 hours), driving after controlled habitual sleep (8.5 hours) (Road 1), and driving after reduced sleep (2 hours) (Road 2). SETTING: Sleep laboratory and open French highway. PARTICIPANTS: 10 male participants (mean age 22 years, range 18-24 years, mean driving distance per year 15000 Km/9000 miles) free of sleep disorders. MEASUREMENTS: Simple reaction time, prospective self-assessment of performance, and instantaneous fatigue and sleepiness ratings measured at 2-hour intervals. RESULTS: A two-way repeated ANOVA with time of day and condition indicated a significant main effect for time of day (p < 0.05). The interaction between the two factors (condition * time of day) was also significant (p < 0.05). The effects of time of day were significant only in the condition of driving after sleep restriction, (p < 0.05). Under sleep restriction, some drivers presented an increase of 650 milliseconds compared to the laboratory condition, representing an increase of 23 meters in breaking distance at a speed of 75 miles per hour. Correlation analyses showed a significant linear correlation between self-assessment and reaction time in the laboratory condition (r = -0.58, p < 0.01) but not in the road conditions. Self ratings during the breaks showed a significant increase in instantaneous self rated fatigue and sleepiness between Road 1 and Road 2 conditions (Wilcoxon's test, Z = - 6.47, p < 0.0001 and Z = - 6.26, p < 0.0001). CONCLUSIONS: Sleep restriction combined with fatigue significantly affects reaction time. The lack of correspondence between reaction time and prospective self-evaluation of performance suggests that self-monitoring in real conditions is poorly reliable. PMID- 12749547 TI - Decreased hypocretin-1 (Orexin-A) levels in the cerebrospinal fluid of patients with myotonic dystrophy and excessive daytime sleepiness. AB - STUDY OBJECTIVE: Myotonic dystrophy type 1 is a multisystem disorder with myotonia, muscle weakness, cataracts, endocrine dysfunction, and intellectual impairment. This disorder is caused by a CTG triplet expansion in the 3' untranslated region of the DMPK gene on 19q13. Myotonic dystrophy type 1 is frequently associated with excessive daytime sleepiness, sharing with narcolepsy a short sleep latency and the presence of sleep-onset rapid eye movement periods during the Multiple Sleep Latency Test. Since narcolepsy is characterized by a dysfunction of the hypothalamic hypocretin system, we investigated whether patients with myotonic dystrophy type 1 with excessive daytime sleepiness have abnormalities in the hypocretin system. DESIGN/PARTICIPANTS: Six patients with myotonic dystrophy type 1 complaining of excessive daytime sleepiness and 13 healthy controls without a sleep disorder were included. The patients with myotonic dystrophy type 1 were evaluated using clinical interviews, nocturnal polysomnograms, and Multiple Sleep Latency Tests. All patients had a confirmed genetic diagnosis for DM1 and were HLA typed. Cerebrospinal fluid hypocretin-1 levels were measured using a direct radioimmunoassay in patients and controls. SETTING: University hospital sleep laboratory. INTERVENTIONS: N/A. MEASUREMENT AND RESULTS: The mean sleep latency on Multiple Sleep Latency Tests was abnormal in all patients (< 5 minutes in 2, < or = 8 in 4) and 2 sleep-onset rapid eye movement periods were observed in 2 subjects. All patients were HLA-DQB1*0602 negative. Hypocretin-1 levels were significantly lower in patients versus controls (p < 0.001); 1 case with 2 sleep-onset rapid eye movement periods had hypocretin-1 levels in the range generally observed in narcolepsy (< 110 pg/mL). Three cases had intermediate levels (110-200 pg/mL). Hypocretin-1 levels did not correlate clinically with disease severity or duration or with subjective or objective sleepiness reports. CONCLUSIONS: A dysfunction of the hypothalamic hypocretin system may mediate sleepiness and abnormal Multiple Sleep Latency Test results in patients with myotonic dystrophy type 1. PMID- 12749548 TI - Relationship of sleep apnea to functional capacity and length of hospitalization following stroke. AB - STUDY OBJECTIVES: Recent evidence indicates that sleep apnea is common in patients with stroke. We hypothesized that the presence of sleep apnea among stroke patients would be associated with a greater degree of functional disability and longer hospitalization following stroke. DESIGN: Prospective study. SETTING AND PATIENTS: Sixty-one stroke patients admitted to a stroke rehabilitation unit. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Sleep studies were performed on all patients, and sleep apnea was defined as an apnea-hypopnea index of 10 or more per hour of sleep. Patients underwent functional assessments, including the Functional Independence Measure. Sleep apnea was found in 72% of patients; 60% had predominantly obstructive sleep apnea, while 12% had predominantly central sleep apnea. Although the severity of stroke was similar in the 2 groups, compared to patients without sleep apnea, those with sleep apnea had lower functional capacity [Functional Independence Measure score (mean +/- SEM) 80.2 +/- 3.6 versus 94.7 +/- 4.3, p < 0.05 at admission, and 101.5 +/- 2.8 versus 112.9 +/- 2.7, p < 0.05 at discharge] and spent significantly more days in rehabilitation (45.5 +/- 2.3 versus 32.1 +/- 2.7 days, p < 0.005). In addition, multiple regression analysis showed that obstructive sleep apnea was significantly and independently related to functional impairment and length of hospitalization. CONCLUSIONS: Sleep apnea is very common among stroke patients undergoing rehabilitation, and its presence is associated with worse functional impairment and a longer period of hospitalization and rehabilitation. These data suggest that sleep apnea may be contributing to functional impairment and prolonged hospitalization following stroke. PMID- 12749549 TI - The neuropsychological effects of obstructive sleep apnea: a meta-analysis of norm-referenced and case-controlled data. AB - STUDY OBJECTIVES: The research literature on the neuropsychological effects of obstructive sleep apnea (OSA) has yielded seemingly contradictory findings, and narrative reviews of this literature are prone to interpretive errors. We used sophisticated meta-analytic models to minimize such errors, with the goal of clarifying the effect of OSA on neuropsychological functioning. DESIGN: Meta analytic review of research through 2001. PARTICIPANTS: We reviewed studies of neuropsychological functioning among adults with untreated OSA. Twenty-five studies met review criteria, representing 1092 patients with OSA and 899 healthy controls. MEASUREMENTS AND RESULTS: Two sets of effect sizes were generated. One compared OSA group means against those of healthy controls in case-controlled studies. The other compared all OSA group means against published normative data. Within each data set, 10 neuropsychological outcome domains were coded. In both data sets, untreated OSA was found to have a negligible impact on intellectual and verbal functioning but a substantial impact upon vigilance and executive functioning. Data were mixed with regard to visual and motor functioning; posthoc inspection of the data suggested that tests of fine-motor coordination or drawing were more sensitive to OSA than were tests of fine-motor speed or visual perception. Data were also mixed with regard to memory functioning, probably related to methodologic differences across studies. CONCLUSIONS: Etiologic models should emphasize mechanisms known to affect vigilance, executive functioning, and motor coordination but not intelligence, verbal functioning, or visual perception. Clinicians should be alert to OSA symptoms in patients with declines in vigilance, executive functioning, or coordination. PMID- 12749550 TI - Sleep during titration predicts continuous positive airway pressure compliance. AB - STUDY OBJECTIVES: Poor compliance with continuous positive airway pressure (CPAP) has been identified as a significant obstacle in the treatment of obstructive sleep apnea. While previous studies have focused on diagnostic screening variables, side effects, health beliefs, and measures of disease severity, investigators have generally ignored sleep parameters assessed during CPAP titration as predictors of compliance. As the titration night represents patients' initial exposure to nocturnal CPAP treatment, we hypothesized that nocturnal polysomnographic (PSG) variables, representing improved sleep at this time, would predict higher subsequent compliance. DESIGN: Prospective analyses of a sequential case series were undertaken using nocturnal PSG variables during titration as early predictors of CPAP compliance. SETTING: Accredited sleep center. PATIENTS: Seventy-one patients with sleep apnea, aged 31-78 years, with a mean respiratory disturbance index of 62.0 +/- 32.2. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Compliance was calculated as mean hours per night of CPAP use over the initial follow-up period (mean 46.9 days). Standard PSG variables and subjective reports of sleep were used as predictive variables in multivariate analyses. Mean objective compliance was 5.04 hours per night +/- 2.59. Consistent with our hypothesis, the best predictor of compliance was change in sleep efficiency (SE) from diagnostic to titration night [F (1,66) = 17.31, p < .000 (r = .48)], indicating that patients whose sleep improved most on the titration night had the highest levels of compliance. This relationship was also significant after controlling for measures of disease severity obtained during the diagnostic testing night. Importantly, individuals whose sleep improved on the CPAP titration night had nightly compliance rates of approximately 2 hours greater than patients whose sleep did not improve during titration. CONCLUSIONS: The findings suggest that patients' initial experience with CPAP treatment and, in particular, the degree of improvement in sleep during CPAP titration may be crucial factors in determining their subsequent use of this treatment modality. PMID- 12749551 TI - Sleep EEG power spectra, insomnia, and chronic use of benzodiazepines. AB - STUDY OBJECTIVES: The present study examined the sleep microstructure in relation to insomnia and chronic use of benzodiazepines in older adults. PARTICIPANTS: The participants were 46 older adults, aged 55 or older (mean age = 62.9), who were divided into three groups: insomnia sufferers using BZ chronically (n = 15), drug free insomnia sufferers (n = 15), and self-defined good sleepers (n = 16). DESIGN: Participants completed 3 consecutive nights of polysomnography in the laboratory. Spectral analyses were carried on the second night of sleep recordings. Stages 2, 3, and 4 of the first 4 cycles of the second night were retained for the analysis. RESULTS: Results showed no significant differences between drug-free insomnia sufferers and good sleepers. However, benzodiazepine users exhibited significantly less delta and theta activity over the night than did good sleepers. When compared to drug-free insomnia sufferers, benzodiazepine users had less delta and theta activity within cycle 2 only. Regarding high frequency bands, benzodiazepine users had more beta 1 activity within cycle 3 than did good sleepers and more than both drug-free insomnia sufferers and good sleepers within cycle 4. CONCLUSIONS: The findings show that spectral analysis is an efficient tool to detect and quantify the effects of benzodiazepine use on sleep structure, particularly with older adults, a group for whom macrostructure sleep alterations due to physiologic aging are hard to distinguish from sleep changes induced by insomnia and the use of hypnotic drugs. In addition, these results raise important questions about the effects and indications of prolonged use of benzodiazepine medications in older adults with insomnia complaints. PMID- 12749552 TI - Sleep disorders in the elderly with and without chronic airflow obstruction: the SARA study. AB - OBJECTIVE: Our objectives were to test the hypothesis that, in the geriatric population, chronic airway obstruction is associated with a higher prevalence of sleep disturbances; to identify the main correlates of sleep disturbances, and to verify whether asthma and COPD patients have different patterns of sleep disturbances. METHODS: The EPESE questionnaire was administered to 734 patients aged 65 years and over with asthma or chronic obstructive pulmonary disease (cases) and 1237 individuals of comparable age who were free of respiratory disease but not of other chronic conditions (controls). Four sleep disturbances were quantified: difficulty in falling asleep, nocturnal awakening, morning tiredness, and early awakening. Multidimensional assessment of demographic data, personal history, clinical, and functional status was performed. Independent correlates of sleep disturbances were identified by logistic regression analysis. RESULTS: One or more sleep disturbances were reported by 445 cases and 697 controls (60.6% vs. 56.4%, ns). Morning tiredness and early awakenings were more prevalent among cases (38% vs. 27.8%, p < 0.001, and 35.1% vs. 28%, p < 0.001, respectively). Depression, as assessed by the 15-item Geriatric Depression Scale, was the most significant independent correlate for all sleep scores. Both being a case and having arthritis were independent correlates of three out of the four sleep disturbances. CONCLUSIONS: Selected sleep disturbances are more common among elderly patients with chronic airway diseases than in those with chronic non-respiratory diseases. Depressed mood and coexisting arthritis are the most relevant independent correlates of sleep disturbances. PMID- 12749553 TI - Comparison of subjective and objective measures of insomnia in monozygotic twins discordant for chronic fatigue syndrome. AB - STUDY OBJECTIVES: To examine the objective and subjective measures of insomnia in chronic fatigue syndrome (CFS). DESIGN: Monozygotic co-twin control study. SETTING: Academic medical center. PATIENTS OR PARTICIPANTS: Twenty-two pairs of monozygotic twins where 1 member of the pair had CFS and the other did not. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Twenty-two CFS-discordant twin pairs completed a Sleep Disorders Questionnaire, overnight polysomnography, and a postpolysomnography sleep survey. Mean and percent differences in the sleep measures were compared between the CFS and healthy twins using matched-pair methods of analysis. Compared with their healthy co-twins, the CFS twins more frequently endorsed 8 subjective measures of insomnia and poor sleep (all p < or = 0.05). However, the CFS and healthy twins did not differ in objective polysomnographic measures of insomnia, including sleep latency, total sleep time, sleep efficiency, arousal number, arousal index, hypnogram awakenings, rapid eye movement (REM)-sleep latency, and percent stages 1, 2, and 3-4 (delta). Percent stage REM sleep was increased in the CFS twins compared with the healthy twins (27.7% vs. 24.4%, p < or = 0.05). On the postpolysomnography survey, CFS twins reported that they had slept fewer hours (6.2 vs. 6.7; p < or = 0.05), and were less well rested (p < or = 0.001) compared to their co-twins. CONCLUSIONS: CFS patients had worse subjective sleep than their co-twins despite little objective data supporting this discrepancy, suggesting they suffer from an element of sleep state misperception. The higher percentage of REM sleep in the CFS twins implies that REM sleep may play a role in this illness. PMID- 12749554 TI - Effect of ethnicity on sleep: complexities for epidemiologic research. AB - STUDY OBJECTIVES: The goal of this study was to examine whether there were ethnic differences in polysomnographically recorded sleep, either in the controlled laboratory environment or in the home setting. DESIGN: Prospective study of ethnic differences in stress physiology and sleep. SETTING: Two sleep recordings were performed on consecutive nights in a hospital-based sleep laboratory, followed 1 to 4 weeks later by a third sleep recording in the subject's home. PARTICIPANTS: 51 employed healthy adult subjects, aged 15 to 50 years. 24 self identified as black, and 27 as white. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: Blacks had less slow wave sleep than did whites in both the sleep laboratory and in the home. Blacks had significantly more slow wave sleep at home compared to the hospital setting, while the reverse was true for whites. This location-by-ethnicity interaction could not be accounted for by depression ratings or social class. CONCLUSIONS: The home setting is generally considered to be more ecologically valid than the controlled hospital-based laboratory setting for the monitoring of sleep. These data suggest that ethnicities may respond differentially to the sleeping environment. This observation may need to be taken into account in future epidemiologic studies of sleep. PMID- 12749555 TI - Competency-based goals for sleep and chronobiology in undergraduate medical education. AB - STUDY OBJECTIVES: Sleep and circadian rhythms are biologic processes operative in health and disease, but as yet there is no articulated curriculum for undergraduate medical education. DESIGN: A multidisciplinary expert-opinion approach was utilized to assess and define education objectives and the potential for implementation. SETTING: N/A. PATIENTS OR PARTICIPANTS: National Institutes of Health Sleep Academic Awardees. INTERVENTIONS: N/A. RESULTS: Four competencies with examples of instruction objectives were identified relating to sleep processes and sleep need, the impact of sleep and sleep disorders on human illness, the sleep history, and the application of sleep physiology and pathophysiology to patent care. Various strategies and tools are currently available for implementation and assessment of learning objectives for these knowledge and skills. CONCLUSION: The core competencies can be designed to improve physician knowledge and skills in recognizing and intervening in sleep problems and disorders. Learning objectives can be immediately incorporated into most medical school curricula. At the same time, these competencies serve as an important bridge across multiple medical content areas and disciplines and between undergraduate and postgraduate training. PMID- 12749556 TI - Practice parameters for the role of actigraphy in the study of sleep and circadian rhythms: an update for 2002. AB - Actigraphy is a method used to study sleep-wake patterns and circadian rhythms by assessing movement, most commonly of the wrist. These evidence-based practice parameters are an update to the Practice Parameters for the Use of Actigraphy in the Clinical Assessment of Sleep Disorders, published in 1995. These practice parameters were developed by the Standards of Practice Committee and reviewed and approved by the Board of Directors of the American Academy of Sleep Medicine. Recommendations are based on the accompanying comprehensive review of the medical literature regarding the role of actigraphy, which was developed by a task force commissioned by the American Academy of Sleep Medicine. The following recommendations serve as a guide to the appropriate use of actigraphy. Actigraphy is reliable and valid for detecting sleep in normal, healthy populations, but less reliable for detecting disturbed sleep. Although actigraphy is not indicated for the routine diagnosis, assessment, or management of any of the sleep disorders, it may serve as a useful adjunct to routine clinical evaluation of insomnia, circadian-rhythm disorders, and excessive sleepiness, and may be helpful in the assessment of specific aspects of some disorders, such as insomnia and restless legs syndrome/periodic limb movement disorder. The assessment of daytime sleepiness, the demonstration of multiday human-rest activity patterns, and the estimation of sleep-wake patterns are potential uses of actigraphy in clinical situations where other techniques cannot provide similar information (e.g., psychiatric ward patients). Superiority of actigraphy placement on different parts of the body is not currently established. Actigraphy may be useful in characterizing and monitoring circadian rhythm patterns or disturbances in certain special populations (e.g., children, demented individuals), and appears useful as an outcome measure in certain applications and populations. Although actigraphy may be a useful adjunct to portable sleep apnea testing, the use of actigraphy alone in the detection of sleep apnea is not currently established. Specific technical recommendations are discussed, such as using concomitant completion of a sleep log for artifact rejection and timing of lights out and on; conducting actigraphy studies for a minimum of three consecutive 24 hour periods; requiring raw data inspection; permitting some preprocessing of movement counts; stating that epoch lengths up to 1 minute are usually sufficient, except for circadian rhythm assessment; requiring interpretation to be performed manually by visual inspection; and allowing automatic scoring in addition to manual scoring methods. PMID- 12749557 TI - The role of actigraphy in the study of sleep and circadian rhythms. AB - In summary, although actigraphy is not as accurate as PSG for determining some sleep measurements, studies are in general agreement that actigraphy, with its ability to record continuously for long time periods, is more reliable than sleep logs which rely on the patients' recall of how many times they woke up or how long they slept during the night and is more reliable than observations which only capture short time periods. Actigraphy can provide information obtainable in no other practical way. It can also have a role in the medical care of patients with sleep disorders. However, it should not be held to the same expectations as polysomnography. Actigraphy is one-dimensional, whereas polysomnography comprises at least 3 distinct types of data (EEG, EOG, EMG), which jointly determine whether a person is asleep or awake. It is therefore doubtful whether actigraphic data will ever be informationally equivalent to the PSG, although progress on hardware and data processing software is continuously being made. Although the 1995 practice parameters paper determined that actigraphy was not appropriate for the diagnosis of sleep disorders, more recent studies suggest that for some disorders, actigraphy may be more practical than PSG. While actigraphy is still not appropriate for the diagnosis of sleep disordered breathing or of periodic limb movements in sleep, it is highly appropriate for examining the sleep variability (i.e., night-to-night variability) in patients with insomnia. Actigraphy is also appropriate for the assessment of and stability of treatment effects of anything from hypnotic drugs to light treatment to CPAP, particularly if assessments are done before and after the start of treatment. A recent independent review of the actigraphy literature by Sadeh and Acebo reached many of these same conclusions. Some of the research studies failed to find relationships between sleep measures and health-related symptoms. The interpretation of these data is also not clear-cut. Is it that the actigraph is not reliable enough to the access the relationship between sleep changes and quality of life measures, or, is it that, in fact, there is no relationship between sleep in that population and quality of life measures? Other studies of sleep disordered breathing, where actigraphy was not used and was not an outcome measure also failed to find any relationship with quality of life. Is it then the actigraph that is not reliable or that the associations just do not exist? The one area where actigraphy can be used for clinical diagnosis is in the evaluation of circadian rhythm disorders. Actigraphy has been shown to be very good for identifying rhythms. Results of actigraphic recordings correlate well with measurements of melatonin and of core body temperature rhythms. Activity records also show sleep disturbance when sleep is attempted at an unfavorable phase of the circadian cycle. Actigraphy therefore would be particularly good for aiding in the diagnosis of delayed or advanced sleep phase syndrome, non-24-hour-sleep syndrome and in the evaluation of sleep disturbances in shift workers. It must be remembered, however, that overt rest-activity rhythms are susceptible to various masking effects, so they may not always show the underlying rhythm of the endogenous circadian pacemaker. In conclusion, the latest set of research articles suggest that in the clinical setting, actigraphy is reliable for evaluating sleep patterns in patients with insomnia, for studying the effect of treatments designed to improve sleep, in the diagnosis of circadian rhythm disorders (including shift work), and in evaluating sleep in individuals who are less likely to tolerate PSG, such as infants and demented elderly. While actigraphy has been used in research studies for many years, up to now, methodological issues had not been systematically addressed in clinical research and practice. Those issues have now been addressed and actigraphy may now be reaching the maturity needed for application in the clinical arena. PMID- 12749558 TI - Design and implementation of a modulator-based free-space optical backplane for multiprocessor applications. AB - Design and implementation of a free-space optical backplane for multiprocessor applications is presented. The system is designed to interconnect four multiprocessor nodes that communicate by using multiplexed 32-bit packets. Each multiprocessor node is electrically connected to an optoelectronic VLSI chip which implements the hyperplane interconnection architecture. The chips each contain 256 optical transmitters (implemented as dual-rail multiple quantum-well modulators) and 256 optical receivers. A rigid free-space microoptical interconnection system that interconnects the transceiver chips in a 512-channel unidirectional ring is implemented. Full design, implementation, and operational details are provided. PMID- 12749559 TI - Holographic diffuser by use of a silver halide sensitized gelatin process. AB - Diffusers play an important role in liquid crystal display (LCD) application as a beam-shaping device, a brightness homogenizer, a light-Scattering device, and an imaging screen. The transmittance and diffusing angle of the diffusers are the critical aspects for the applications to the LCD. The holographic diffusers by use of various processing methods have been investigated. The diffusing characteristics of different diffusing materials and processing methods have been evaluated and compared. The microstructures of holographic diffuse have been investigated by use of using scanning electron microscopy. The holographic diffusers by use of the silver halide sensitized gelatin (SHSG) method have the structural merits for the improvement of the quality of diffusers. The features of holographic diffuser were exceptional in terms of transmittance and diffusing angle. The replication method by use of the SHSG process can be directly used for the manufacturing of diffusers for the display application. PMID- 12749560 TI - Noniterative two-dimensional phase-retrieval method from two Fourier intensities by use of an exponential filter. AB - A noniterative method of retrieving the two dimensional phase of a wave field from two intensity measurements is proposed. In the measurements, one records two far-field intensities of the wave field modulated and unmodulated with an exponential filter. The phase retrieval method is based on the solution of the simultaneous equations with unknown coefficients of the two-dimensional discrete Fourier transform for the phase. Then there is no need for the information about the wave field, which is used in iterative phase-retrieval methods. The usefulness of this method is shown in computer-simulated examples of the reconstruction of two-dimensional complex amplitude objects. PMID- 12749562 TI - Analysis of illumination coherence properties in small-source systems such as synchrotrons. AB - Modern synchrotron beamlines often take the form of critical illumination systems, where an incohrent source of limited spatial extent is re-imaged to an experimental plane of interest. Unique constraints of synchrotron sources and beamline, however, may preclude the use of the simple Zernike approximation for calculating the object-image coherence relationship. Here, we perform a rigorous analysis of the object-image coherence relationship valid for synhrotron beamlines. The analysis shows that beamline aberrations have an effect on the coherence properties. Effects of various low-order aberrations on the coherence properties are explicitly studied. PMID- 12749561 TI - Phase unwrapping by region growing. AB - The phase unwrapping problem consists in mingling out an integer field whose values make the original wrapped phase field continuous. Even if in principle the problem is very simple--a direct integration of the wrapped phase field suffices- in the presence of noise and/or undersampling, the solution is no longer unique and the direct integration methods usually fail to find an acceptable solution. This work presents what is to my knowledge a new unwrapping algorithm that attempts to find the solution by iteratively merging and shifting the continuous areas until a single region is built or no further moves are possible. Unlike the tile methods, the regions can have arbitrary shape and need not be ingle connected so that, by removing the predefined size and shape constraint, the algorithm is very robust. The greater freedom of the regions' shape makes their handling more problematic, so that certain implementation aspects, critical to algorithm performance, are presented here. Some unwrapping examples are also presented and memory requirements are discussed. PMID- 12749563 TI - Wide-viewing integral three-dimensional imaging by use of orthogonal polarization switching. AB - A wide-viewing integral three-dimesional (3D) imaging system that adopts orthogonal polarization switching is proposed and demonstrated. In our scheme,the polarizing sheet attached to the lens array and the orthogonal polarization switching of the elemental image array perform elemental lens switching. The experimental results document that the viewing angle becomes remarkably wider than that of the conventional method. The distinguishing feature of our system is that it requires no mechanical moving part. In addition, because a commercially available polarization shutter screen is used for electrical switching, it is easy to implement this as a practical system. We believe that the proposed method facilitates the practical use of this wide-viewing integral 3D imaging system. PMID- 12749564 TI - Comment: absolute measurement of roughness and lateral-correlation length of random surfaces by use of the simplified model of image-speckle contrast. AB - This comment demonstrates several errors in the derivation of speckle contrast and corresponding measurements reported by Cheng et at. (Appl. Opt. 41, 4148 (2002)] for a 4f optical system. In particular, the theoretical derivation is wrong: It is used outside of its domain of validity, and the experimental results do not support the theoretical analysis. PMID- 12749566 TI - Analysis and evaluations of logical instructions called in parallel digital optical operations based on optical array logic. AB - The authors have analysed and evaluated a two-dimensional instruction set of parallel operation based on optical array logic (OAL), which is a digital optical computing paradigm, to clarify efficient composition of an optical computing system based on OAL. To evaluate parallel operation based on OAL, the author have introduced new indices and evaluated a logical instruction set of various parallel operations with the indices, so that a guideline for composing a simple and efficient OAL computing system is clarified. Also, the authors have proposed the reduced operation kernel set correlation technique to perform parallel operations more efficiently by a simple OAL computing system. It has been clarified that the technique can reduce the required hardware necessary for an OAL computing system for efficient general-purpose processing. PMID- 12749565 TI - Laser excess noise reduction in optical phase-shift measurements. AB - We describe an optical detection scheme that reduces laser excess noise in measurements of small optical phase shifts. The scheme improves the sensitivity of the balanced differential detection. Analysis of the scheme and comparison with the conventional detection are presented. The scheme is applied in electro optic probing of electrical signals in integrated circuits, where the excess laser noise is reduced by approximately 20 dB. PMID- 12749567 TI - Experimental demonstration of gray-scale sparse modulation codes in volume holographic storage. AB - We discuss experimental results of a versatile nonbinary modulation and channel code appropriatefor two-dimentional page-oriented holographic memories. An enumerative permutation code is used to provide a modulation code that permits a simple maximum-likelihood detection scheme. Experimental results from the IBM Demon testbed are used to characterize the performance and feasibility of the proposed modulation and channel codes. A reverse coding technique is introduced to combat the effects of error propagation on the modulation-code performance. We find experimentally that level-3 pixels achieve the beet practical result, offering an 11-35% improvement in capacity and a 12% increase in readout rate as compared with local binary thresholding techniques. PMID- 12749568 TI - Perceived lack of ADEA House of Delegates resolutions that focused on policy issues. PMID- 12749569 TI - Reexamining educational philosophy: the issue of professional responsibility, "Cleveland First". AB - This paper proposes a shift of emphasis in the dental curriculum from measures to protect and improve the oral health of individuals to measures to protect and improve the oral health of the community or society. This shift represents a fundamental change in educational philosophy of the dental school. To illustrate this shift in emphasis, this paper describes a demonstration project to test the feasibility of this approach involving all seventy first-year students in the Case Western Reserve University School of Dentistry in a four-week experience placing dental sealants in erupting molars of second and sixth graders in fifty schools of the Cleveland City School System. In future years, the program is expected to reach all second and sixth graders in the Cleveland School System. The experience is a required integral component of the curriculum, involving every student in the class, and is designed to make a demonstrable difference in oral health in the City of Cleveland. The experience is reinforced with course material on professional responsibility. The school is developing additional intensive experiences for second-, third-, and fourth-year classes involving smoking prevention for adolescents, oral health maintenance for nursing home residents, and dental care delivery in the inner city. The initial year of the program has had effects on students' responses to other elements of the first year curriculum that go beyond the experience of placing sealants in children's teeth. The focused efforts of dental students every year are expected to have a measurable effect on the disparities in oral health found in the City of Cleveland as well as a measurable effect on dental students' and dentists' attitudes concerning professional responsibility. PMID- 12749570 TI - A comparison of private and public dental students' perceptions of extramural programming. AB - This project was undertaken to compare the opinions of private and public dental school students' perceptions concerning extramural programming, which is defined as any aspect of the curriculum in which undergraduate dental students provide dental care outside the main dental facility. A survey instrument was used to collect data from undergraduate students at a private (N = 267; 88.4 percent response rate) and at a public (N = 213; 67.2 percent response rate) dental school. When asked to rate the value of various extramural sites in making them a better dentist, both groups rated private dental offices the most valuable and prisons the least valuable. When questioned about the amount of time students should spend each year in extramural programming, private students, overall, desired 34 percent more time than did public students. When asked what percentage of the total time spent in extramural programming students should spend providing various categories of dental care, public school students thought 26 percent more time should be spent rendering preventive services/health education than did the private students. The private students indicated a stronger desire (13 percent more) for rendering clinical services than did public students. Both private and public students were most likely to enter group private practice after graduation. The increasing interest in community-based programs makes the information gained from this study useful for future curriculum planning. PMID- 12749571 TI - Alumni perspectives on community-based and traditional curricula. AB - In 1994, the University of Medicine and Dentistry of New Jersey-New Jersey Dental School (UMDNJ-NJDS) launched the Community-Oriented Dental Education (CODE) program. The CODE program provides senior dental students the opportunity to spend four days per week providing dental care in a community-based clinic. A survey of graduates of CODE (n = 55) and randomly selected graduates of the traditional curriculum (n = 110) was conducted via mail to determine attitudes relating to community service (CS), community-based learning (CBL), reasons for participating in their clinical program, perceived levels of clinical preparedness at graduation, and practice choices. A total of 111 surveys (66.9 percent) were returned to NJDS, with 84.6 percent of CODE alumni responding and 59.0 percent of traditional alumni (TA) responding. Of the 111 surveys returned, sixty-five (58.6 percent) were completed by TA, and forty-six (41.4 percent) were completed by CODE alumni. There were no differences among CODE and TA regarding attitudes toward CS and tendency to practice in underserved areas or to accept Medicaid payments. There were, however, some differences in attitudes toward CBL, reasons for applying or not applying to the CODE program, perceived impact of clinical education on graduates' preparedness, views of the extent to which the programs encouraged students to choose public or private areas of practice, and perceptions of how the desire to help communities influenced career and practice decisions. Some of these findings may be useful to schools as they plan extramural education programs. PMID- 12749572 TI - Distance education in the U.S. and Canadian undergraduate dental curriculum. AB - A major trend at all levels of education in recent years has been the advent of distance learning and, more specifically, the use of computers and communications capabilities to provide online learning. The purpose of this study was to determine the extent to which distance learning and online technologies are being employed by dental schools in the United States and Canada. Two groups were surveyed: academic deans and faculty members of U.S. and Canadian dental schools. Thirty-eight academic deans responded to a paper-based survey, and more than 400 faculty members responded to a web-based survey. The results of these surveys indicate that online delivery of content and information has a bright future in the delivery of the dental school curriculum. At the same time, formidable obstacles must be addressed for this approach to be successful. PMID- 12749573 TI - The effects of teaching dental students brief motivational interviewing for smoking-cessation counseling: a pilot study. AB - The purpose of this research was to explore the changes that occurred in dental students' counseling techniques as a result of training in Brief Motivational Interviewing (BMI). A randomized pretest, posttest design was used with eleven students in each group. Baseline and posttraining measures of students' counseling techniques and the characteristics of the counseling session with standardized patients were made. The measures were: student behavior from videotapes, patient involvement in the treatment, establishment of good doctor patient rapport, perceived efficacy in promoting patient change, and student confidence and interest in the task. MANOVA found significant differences between the trained and the untrained groups (F = 4.019, p = 0.018). Training resulted in sessions in which students used more BMI techniques and patients were more actively involved. No changes were seen in the other variables. Future studies must examine whether more experience will improve the students' ability to use BMI to enhance patient rapport, to increase their sense of competence and interest in doing counseling, and to determine the effectiveness of the counseling to encourage patient smoking cessation. PMID- 12749575 TI - Diabetes mellitus as a modulating factor of endodontic infections. AB - Diabetes mellitus is a chronic disease with serious health consequences. The association between diabetes and periodontal disease is well documented. However, the progression and healing of endodontic infections in diabetic patients has not been adequately studied. In this review, diabetes mellitus is explored as a potential modulating factor of endodontic pathosis. Recent data on the relationship between the clinical presentation of pulpal and periradicular disease, as well as the outcome of endodontic treatment in diabetic and nondiabetic patients, are presented. Diabetics who present for endodontic treatment, particularly those with periradicular pathosis, may have increased perioperative symptoms. Cases with preoperative periradicular lesions are less likely to be determined successful two years or longer postoperatively if the patient reports a history of diabetes. Studies examining the pathogenesis of periradicular lesions in mouse models with uncontrolled type 1 diabetes suggest that the lesion size may be increased and the animals have increased serious sequelae. Preliminary findings suggest that some bacterial species may be more prevalent in necrotic pulp of diabetic than nondiabetic patients. More studies are needed to further explore the microbiology of endodontic infections and to determine effective treatment strategies in both diabetic and nondiabetic patients. PMID- 12749574 TI - Systematic review of controlled trials on the effectiveness of fluoride gels for the prevention of dental caries in children. AB - Fluoride gels have been widely used since the 1970s. The aim of this review was to assess the effectiveness and safety of fluoride gels in the prevention of dental caries in children and to examine factors potentially modifying their effectiveness. Relevant randomized or quasi-randomized trials were identified without language restrictions by searching multiple databases, reference lists of articles, and journals and by contacting selected authors and manufacturers. Trials with blind outcome assessment comparing fluoride gel with placebo or no treatment for at least one year and involving children under seventeen years of age were selected. Inclusion decisions, quality assessment, and data extraction were duplicated in a random sample of one third of studies, and consensus was achieved by discussion or a third party. Random effects meta-analyses were performed where data could be pooled. Potential sources of heterogeneity were examined in random effects meta-regression analyses. The main outcome was caries increment measured by the change in decayed, missing, and filled permanent tooth surfaces (D(M)FS). The primary measure of effect was the prevented fraction (PF) that is the difference in mean caries increment between the treatment and control groups expressed as a percentage of the mean increment in the control group. Potential adverse effects and unacceptability of treatment were also recorded. Twenty-five studies were included, involving 7,747 children. For the twenty-three that contributed data for meta-analysis, the D(M)FS pooled prevented fraction estimate was 28 percent (95 percent CI, 19 percent to 37 percent; p < 0.0001). There was clear heterogeneity, confirmed statistically (p < 0.0001). The effect of fluoride gel varied according to type of control group used, with D(M)FS PF on average being 19 percent (95 percent CI, 5 percent to 33 percent; p < 0.009) higher in non-placebo controlled trials. Only two trials reported on adverse events. There is clear evidence of a caries-inhibiting effect of fluoride gel. The best estimate of the magnitude of this effect, based on the fourteen placebo controlled trials, is a 21 percent reduction (95 percent CI, 14 to 28 percent) in D(M)FS. This corresponds to an NNT of two (95 percent CI, 1 to 3) to avoid one D(M)FS in a population with a caries increment of 2.2 D(M)FS/year, or an NNT of twenty-four (95 percent CI, 18 to 36) based on an increment of 0.2 D(M)FS/year. However, further work is needed to identify and quantify potential harmful effects of fluoride gels. PMID- 12749576 TI - Affirmative action: essential to achieving justice and good health care for all in America. AB - Affirmative action is an established principle that brings fairness and justice to admissions policies and practices by setting goals that encourage and pressure institutions and individuals to create educational and professional opportunities for minorities and women, if it were not for affirmative action, we would waste the talents of countless individuals who would be discounted because they are minorities or women. The result would be a nation that is weaker because it would be segregated once again in a system in which white people and men would have the preponderance of opportunity and authority and in which access would be very limited for minorities and women. It may be time to reframe the argument for affirmative action in language that denotes its benefits to all Americans by increasing access for emerging majority citizens now and in the future. ADEA, academic dentistry, and the dental profession should continue to do everything it can to preserve the policies and practices of affirmative action, especially through the support of the University of Michigan admissions policies as challenged in the cases before the U.S. Supreme Court and in our own practices. PMID- 12749577 TI - Optimization of an extracellular protease of Chrysosporium keratinophilum and its potential in bioremediation of keratinic wastes. AB - Chrysosporium keratinophilum IMI 338142 isolated from a waste site containing organopollutants was studied for its ability to produce extracellular proteases on glucose-gelatin medium. Fungus was observed to be a potent producer of such enzymes. Enzyme secretion was best at 15 days of incubation period at pH 8 and temperature 40 degrees C. Asparagine was repressive to protease expression. No relationship existed between the enzyme yield and increase in biomass. Exogenous sugars suppressed enzyme production in the descending order as follows: glucose > arabinose > maltose > mannose > fructose. The enzyme released showed the ability to decompose two keratin substrates tested. Buffalo skin was the most actively degraded substrate when exogenous glucose was absent. Presence of glucose suppressed both enzyme production and degradation of keratin. However, the rate of keratin degradation was independent of enzyme production. PMID- 12749579 TI - Morphological characterization of in-vitro human hair keratinolysis, produced by identified wild strains of Chrysosporium species. AB - Chrysosporium species were isolated from soil and keratinized material. Primary isolation was performed following the general method of hair baiting on modified Czapek-agar media with washed, defated and sterilized human hair fragments added. Strains were maintained in test tubes of potato dextrose agar at 29 degrees C and cultivated on phytone yeast extract agar at 28 degrees C for 14 days for identification. Isolates were characterized using Van Oorschot's key. Keratinolytic activity was expressed following a subjective scale representing degree/severity of attack upon hair surface and presence of fungal structures observed in substrate. Culture results and characterization methods were effective for soil Chrysosporium strain isolation. A new hair attack mode is described. Of 71 keratinolytic fungal isolates, eight (12%) Chrysosporium species were identified. One keratinolytic Chrysosporium sp. isolate is yet to be identified. PMID- 12749578 TI - Genetic relatedness among Filobasidiella species. AB - The three accepted species of Filobasidiella, F. neoformans, F. depauperata, and F. lutea, are compared morphologically and by molecular analysis. Sequences of the internally transcribed spacer (ITS) and the small subunit (SSU) gene of the ribosomal RNA (rRNA) gene cluster were obtained, and analysed by Neighbor-joining and Maximum parsimony methods. The three species of Filobsidiella are shown to form a single monophyletic clade, rooted by Tremella mesenterica. F. lutea was recovered as a distinct, but closely related taxon with the Filobasidiella clade. This is the first report of DNA sequences from herbarium specimens of F. lutea. PMID- 12749580 TI - Effect of different K+ concentrations on Cryptococcus neoformans phenoloxidase activity. AB - Melanin synthesis in Cryptococcus neoformans, catalyzed by phenoloxidase activity, is one of the oldest virulence factors known. However, until now, the relationship between melanin production in C. neoformans and its virulence has been poorly understood. Among different chemical compounds only Fe3+ and Cu2+ cations enhance the phenoloxidase activity in C. neoformans. A few reports in the literature describe the influence of different cations on C. neoformans phenoloxidase activity, excluding iron. In this study, 13 C. neoformans strains isolated from AIDS patients and 7 from bird droppings (B.D.), were examined in order to clarify the effect of different K+ concentrations on phenoloxidase activity. A new solid and liquid caffeic acid minimal synthetic medium (MSM-CAF) containing only caffeic acid and ferric citrate with different potassium concentrations was used to evaluate C. neoformans phenoloxidase activity. In the MSM-CAF solid medium the degree of brown pigmentation on the agar plates was read on days 1, 2 and 3 of incubation, and the pigmentation of the C. neoformans strains was classed into 5 categories. The brown pigment of the liquid MSM-CAF test tubes were checked after 24 hours of incubation by measuring the optical density (O.D.) at 480 nm. Three C. neoformans AIDS and B.D. strains, randomly chosen, were tested for phenoloxidase activity, according to the modified protocols of Polacheck et al., Torres-Guerrero et al. and Rhodes. According to the results obtained, it has been observed that K+ does not activate the phenoloxidase activity in the C. neoformans AIDS and B.D. strains. In particular, with an increase in potassium concentrations in the MSM-CAF solid and liquid medium, there was a corresponding inhibition of the phenoloxidase activity on both the C. neoformans AIDS and B.D. strains. PMID- 12749581 TI - Incidence of keratinophilic fungi from selected soils of Kerala state (India). AB - One hundred and fifty-eight soil samples were collected from various areas of four districts of Kerala and screened for prevalence of keratinophilic fungi and related dermatophytes. From the positive samples (60.75%), a total of eight genera with 15 species were isolated viz., Arthroderma simii (0.63%), Chrysosporium indicum (20.25%), C. keratinophilum (6.96%). C. lobatum (1.26%), C. pannicola (1.26%), C. tropicum (5.06%), Chrysosporium state of Arthroderma cuniculi (1.26%), Chrysosporium state of Ctenomyces serratus (2.53%), Gymnascella hyalinospora (1.26%), Malbranchea aurantiaca (0.63%) M. fulva (1.26%), Microsporum gypseum complex (12.65%), Pseudogymnoascus roseus (1.26%), Trichophyton mentragrophytes (1.26%), and T. terrestre (3.16%). PMID- 12749582 TI - Record of a fatal oomycotic disease of the fresh water fish Anabas cobojius in India caused by Aplanes braunii. AB - The fresh water fish Anabas cobojius (Hamilton-Buchanan 1822), was found to be parasitised by Aplanes braunii de Bary 1888. It is the first and only report of any Aplanes de Bary to cause disease on any fish. A. braunii was found to be a virulent parasite of A. cobojius causing cotton-wool disease involving the skin and fins. The hyphae also penetrated into the tissues of the infected fish and all of them ultimately died. PMID- 12749583 TI - First human case of nocardiosis caused by Nocardia pseudobrasiliensis in Japan. AB - Nocardia sp. IFM 0896, an actinomycete with biochemical characteristics that differed from Nocardia brasiliensis, was isolated from a 71-year-old Japanese man with a history of tuberculosis and cancer. Although the isolate was tentatively identified as N. brasiliensis, the morphological and physiological characteristics of strain IFM 0896 were different from those of N. brasiliensis IFM 0236T. The results of 16S rRNA gene sequence phylogenies and PCR-RFLP analysis of a heat shock protein revealed that Nocardia sp. IFM 0896 belongs to the species N. pseudobrasiliensis. This is the first clinical isolation report of N. pseudobrasiliensis in Japan. PMID- 12749585 TI - Tropic failure of Phyllactinia corylea contributes to the mildew resistance of mulberry genotypes. AB - Different mulberry genotypes show great variation in their resistance to the powdery mildew Phyllactinia corylea. Conidial germination and hyphal growth of P. corylea on the leaf surface of two susceptible mulberry genotypes, viz., Kanva 2 (K2) and Victory 1 (V1) varieties of Morus indica, and on two resistant species, viz,, M. laevigata and M. serrata were studied by scanning electron microscopy. Conidial germination and growth of germ tubes were normal on all the leaves. The hyphae of P. corylea identify stomata on host leaves by their topographical features to produce the stomatopodia precisely over them. The holes and/or the grooves of stomata appear to provide the signals for the initiation of stomatopodia and similar structures are erratically developed over many local depressions or grooves on leaf surface. The abaxial surface of K2 leaf is smooth without prominent undulations of epidermal cell surface, and the stomata are flush with the leaf surface. Although successful penetration is also achieved on V1 leaf, its slightly undulated surface occasionally provides inaccurate tropic signals to the hyphae, inducing the development of stomatopodia away from the stomata. The leaf surfaces of M. laevigata and M. serrata are very rough with highly sculptured cuticle and abundant epidermal outgrowths. Stomata mostly remain sunken or hidden amidst the cuticular ornamentations and the hyphae fail to recognise the precise signals from them. As the surface architecture of the leaves provides many immense sources of tropic signals, stomatopodia are often produced over local depressions or grooves. In these cases the fungus fails to penetrate the leaf, does not develop beyond 24 h and penetration is rarely achieved on the leaves of the resistant plants. The study indicates that the stimulatory effect of the leaf surface topography of resistant varieties misleads the pathogen from successful penetration, thus contributing to the plant's resistance. PMID- 12749586 TI - Ultrastructure of intercellular hypha and haustorium of the rust fungus, Uromyces euphorbiae. AB - The ultrastructure of intercellular hyphae and dikaryotic haustoria of Uromyces euphorbiae, and the host response to haustorial invasion was investigated. The intercellular hyphae share common characteristics with those of other uredinial stages of rust fungi. Three types of septa were recognized inside the intercellular hypha. This study showed that the extrahaustorial membrane was possibly formed before the development of the haustorium. The periodic acid thiocharbohydrazide-silver proteinate technique showed that the haustorial mother cell wall at the penetration site, and the haustorial wall contained more carbohydrates than other fungal structures. In addition, the neckband, present around the haustorial neck, contains different material from those of the rest of the haustorial neck wall. The close associations of host organelles, such as the nucleus, chloroplasts, mitochondria, endoplasmic reticulum and microtubules, with the haustorium, is described. PMID- 12749584 TI - Ecology of dermatophytes and other keratinophilic fungi in swimming pools and polluted and unpolluted streams. AB - The biodiversity and richness of keratinophilic fungal communities including dermatophytes were assessed in three stream sites and three swimming pools in the Nablus district in Palestine, using hair baiting (HBT) and surface dilution plate (SDP) techniques, over 8- and 6-month periods, respectively. The effect of wastewater effluent and selected ecological factors on these fungi in relation to species diversity and population densities were also considered. Fifty keratinophilic fungal species were recovered from the aquatic habitats studied, of which 42 were recovered from stream sites and 22 from swimming pools. Of these fungi 6 were either dermatophytes (Microsporum gypseum, and Trichophyton mentagrophytes) or dermatophyte related species (Chrysosporium merdarium, Ch. tropicum, Ch. keratinophilum and T. terrestre). The most frequently isolated species in the three pools were Acremonium strictum and Cladosporium cladosporioides, using Sabouraud dextrose agar medium (SDA). The most abundant species were Acr. strictum, and Aspergillus flavus. However, only 4 species were isolated using the SDA medium amended with 5-flurocytosine (5-FC). The most frequent and abundant species in the three stream sites using SDA medium were Geotricum candidum, and Penicillium chrysogenum. The most frequent species in the three sites using the 5-FC medium, was Paecilomyces lilacinus. Using HBT, the most abundant and frequent species in the three stream sites were G. candidum, and Pa. lilacinus, on SDA medium, and Pa. lilacinus, and Gliocladium nigrovirens on the 5-FC medium. The 5-FC medium was more suitable for the isolation of dermatophytes and closely related species than the SDA medium; 6 were recovered on 5-FC, whereas only one on the SDA medium. Variation in the levels of keratinophilic fungal populations from the three stream sites sampled 5 times over an 8-month period, followed comparable fluctuation patterns. Wastewater affected fungal population densities with the highest levels in the un-polluted stream sites, and lowest in the heavily polluted sites. Swimming pools, polluted and un-polluted stream sites were found to be rich in pathogenic and potentially pathogenic fungi. PMID- 12749587 TI - Aflatoxin binders I: in vitro binding assay for aflatoxin B1 by several potential sequestering agents. AB - Nine potential proprietary sequestering agents consisting of 4 activated charcoals, 3 sodium bentonites, a calcium bentonite, and an esterified glucomannan were compared in a novel in vitro assay for aflatoxin B1 (AFB1) binding. Agents were evaluated in 10% methanol prepared as 1% stirred suspensions at pH 3, 7, 10 and pH-unadjusted, with or without AFB1 at 5 microg/ml. All nine agents bound more than 95% of the 5 microg of AFB1 in solution, regardless of pH. The sodium bentonites bound 98, 95, and 98% of the AFB1. The four activated charcoals bound over 99%, the calcium bentonite bound 98%, and the esterified glucomannan bound 97% of the AFB1 in solution. The results suggested that the sequestering agents tested here had sufficient potential to bind AFB1 at pH values commonly found in the gastrointestinal tracts of ruminants and other animals. PMID- 12749588 TI - Cloning and functional expression of an esterase gene in Aspergillus parasitcus. AB - Within the 80 kb aflatoxin pathway gene cluster characterized earlier, and between adhA and norA genes, we have identified an estA gene encoding an esterase from wild type strain Aspergillus parasiticus SRRC 143. The 1,500 bp genomic DNA and 945 bp cDNA sequences were determined for estA. Outside of the aflatoxin pathway gene cluster, an additional copy of the estA gene (named estA2) was also cloned from the same A. parasiticus strain. Comparison of the estA and estA2 sequences showed 9 substitutions within the 314 amino acid residues of their gene products, and no apparent defect was identified in the estA2. The estA gene is a homolog of the stcI gene identified in A. nidulans involved in the biosynthesis of sterigmatocystin and dihydro-sterigmatocystin for the conversion of versiconal hemiacetal acetate to versiconal. Reverse-transcriptase polymerase chain reaction (RT-PCR) experiments demonstrated that the estA is constitutively expressed. And only this estA gene, which is located within the aflatoxin pathway gene cluster, is expressed; no expression of the estA2 gene was detected under both aflatoxin conducive and non-conducive conditions. Possible reasons for the preferential expression of the estA over the estA2 gene have been discussed. PMID- 12749589 TI - Distribution, occurrence and characterization of entomopathogenic fungi in agricultural soil in the Palestinian area. AB - The occurrence of entomopathogenic fungi was investigated in irrigated vegetable fields and citrus orchards soils, over a nine-month period (April-December 1999), using the Galleria bait method (GBM). Entomopathogenic fungi were found to occur in 33.6% of the soil samples studied, with positive samples yielding 70 fungal isolates, belonging to 20 species from 13 genera. Conidiobolus coronatus was the most frequent and abundant entomopathogenic species recovered, comprising 31.4% of the total number of isolates. Soil pH, soil moisture content and the geographical location had minor or no effect on the isolation of entomopathogenic fungi in the fields studied. On the other hand, organic matter content of soil, and vegetation type were found to significantly affect the occurrence of entomopathogenic fungi in soil habitats, with orchard fields yielding larger numbers of isolates than vegetable fields. Using Koch's postulates the pathogenicity of fungal isolates to Galleria larvae was found to range from 16 100% (mortality rate). Isolates of C. coronatus proved to be the most virulent isolates recovered. The effect of media and temperature on mycelial growth rate, conidial production and conidial germination of six entomopathogenic fungal species (C. coronatus, Entomophaga grylli, Erynia castrans, Hirsutella jonesii, Paecilomyces farinosus and Sporodiniella umbellata) was also studied. Mycelial growth rate, spore production and spore germination were significantly affected by media, temperature and isolates. In view of the present results, C. coronatus appears to be a good candidate for pest control in agricultural soils, as it has a wide tolerance to agricultural practices, has frequently been isolated from both vegetable and orchard fields, and is characterized by high mycelial growth rate, conidial production and conidial germination. PMID- 12749590 TI - Peroxisome proliferator-activated receptors: a critical review on endogenous pathways for ligand generation. AB - Lipid mediators can exert their effects by interactions with well-characterised cell surface G-protein-linked receptors. Recently, a group of intracellular receptors have been identified that are activated by a large variety of lipid derived mediators. Amongst these novel targets, the peroxisome proliferator activated receptors (PPARs), a family of three (PPARalpha, beta/delta and gamma) nuclear receptor/transcription factors have become a major area for investigation. PPARs are found throughout the body, where they have diverse roles regulating lipid homeostasis, cellular differentiation, proliferation and the immune response. There is a great interest, therefore, in the roles of PPARs in a variety of pathological conditions, including diabetes, atherosclerosis, cancer and chronic inflammation. Although, a number of naturally occurring compounds can activate PPARs, it has been difficult, as yet, to characterise any of these mediators as truly endogenous ligands. These findings have lead to the suggestion that PPARs may act just as general lipid sensors. Acting as lipid sensors, PPARs may take changes in lipid/fatty acid balance in the diet or local metabolism and translate them to tissue-specific ligands, exerting tissue-specific effects. Using classical pharmacological criteria for endogenous mediator classification we will critically discuss the variety of pathways for putative ligand generation. PMID- 12749591 TI - Chronic exposure to TXA2 increases expression of ROCKI in human myometrial cells. AB - Increased expression of RhoA-associated protein kinase (ROK) in human pregnant myometrial tissue is due to increased expression of the p160ROKI isoform. Expression and proteolysis of p160ROKI was investigated in cultured primary human uterine smooth muscle cells stimulated with the stable thromboxane A2 (TXA2) analogue U46619. Acute exposure to U46619 showed no change in protein expression or cleavage of p160ROKI. Chronic exposure to U46619 resulted in a concentration dependent increase in protein expression of p160ROKI that was inhibited by pre treatment of the cells with the C3-exotoxin. Pre-incubation with the thromboxane receptor antagonist SQ29548 also blocked the U46619-mediated increase in p160ROKI protein expression but at the same time promoted increased proteolysis of pre existing p160ROKI to p130ROKI. Pre-treatment of the cells with the caspase 3 inhibitor Z-DEVD-FMK blocked the cleavage of p160ROKI. These findings show that ROKI is an inducible isoform whose aberrant expression and cleavage needs to be controlled to prevent contractile dysfunction. PMID- 12749592 TI - Differences in positional esterification of 14,15-epoxyeicosatrienoic acid in phosphatidylcholine of porcine coronary artery endothelial and smooth muscle cells. AB - Epoxyeicosatrienoic acids (EETs) are readily incorporated into phospholipids of smooth muscle cells (SMC) and endothelial cells (EC). Incorporation of EETs into intact porcine coronary arteries potentiates EC-dependent relaxation, but not vasorelaxation induced by agents that act solely on SMC. To explore the potential mechanisms responsible for this difference, porcine coronary artery SMC and EC preloaded with [3H]14,15-EET were treated with calcium ionophore A23187. Although the amount of EET incorporated into EC and SMC was similar, A23187 stimulated a five-fold increase in release of radioactivity from EC, but only a 21% increase in release from SMC. Thin layer chromatography (TLC) examination of cell lipids demonstrated that > 70% of the incorporated radioactivity was present in phosphatidylcholine (PC) in both SMC and BC. After treatment of EC PC with PLA2, TLC analysis indicated that approximately equal to 75% of radioactivity was present as free EET, and 25% of radioactivity was present as lyso-PC. Therefore, most of the 14,15-EET was esterified into the sn-2 position of PC in EC. However, in SMC, approximately equal to 70% of radioactivity was present as lyso-PC after PLA2 treatment, indicating that the EET was predominately esterified into the sn 1 position. In contrast, all of the 14,15-EET was esterified into the sn-2 position of PI in both EC and SMC. These results suggest that the preferential incorporation of 14,15-EET into the sn-1 position of PC in SMC may help to explain the greater retention of the compound in SMC, while incorporation into the sn-2 position of PC in EC may facilitate agonist-induced 14,15-EET release and potentiation of EC-dependent porcine coronary artery relaxation. PMID- 12749593 TI - The regulation of arachidonic acid metabolism in human first trimester trophoblast by cyclic AMP. AB - Human trophoblast cells are known to release a range of arachidonic acid metabolites into culture medium, including cyclo-oxygenase, lipoxygenase and epoxygenase products. In this study we investigated the effects of dibutyryl cyclic AMP (db cAMP) on arachidonic acid metabolism in human first trimester trophoblast cells, and also determined the distribution of metabolites between intracellular and extracellular compartments. db cAMP increased intracellular levels of radioactivity within 2 min, and extracellular levels of radioactivity were increased after 30 min. These changes were reflected in increased levels of arachidonic acid metabolites in both compartments, indicating that arachidonic acid was metabolised. db cAMP increased intracellular levels of 5,6 epoxyeicosatrienoic acid (5,6-EpETrE) within 2 min of addition to cultured cells. No changes were detected after 5-10 min, but substantial changes were found 30 min after the addition of db cAMP. The dihydroxyeicosatrienoic acid (DiHETrE) breakdown products also increased with similar kinetics. In contrast, levels of 14,15-EpETrE increased after 5-10 min. PMID- 12749594 TI - Effect of luteinizing hormone (LH), pregnancy specific protein B (PSPB), or arachidonic acid (AA) on ovine endometrium of the estrous cycle or placental secretion of prostaglandins E2 (PGE2) and F2alpha (PGF2alpha) and progesterone in vitro. AB - The objective of this experiment was to determine the effect of AA, LH, or PSPB on secretion of PGE2, PGF2alpha, or progesterone by ovine caruncular endometrium of the estrous cycle or placental tissue of pregnancy in vitro. Ovine caruncular endometrium of the estrous cycle (days 8, 11, 13, and 15) or caruncular/placental tissue on days 8, 11, 13, 15, 20, 30, 40, 50, 60, and 90 postbreeding were incubated in vitro with vehicle, AA, LH, or PSPB in M-199 for 4 and 8 h. Secretion of PGF2alpha by caruncular endometrium of non-bred ewes on days 13 and 15 and by caruncular/placental tissue of bred ewes on days 13, 15, 20, 30, and 40 was increased (P < or = 0.05) when incubated with vehicle and declined (P < or = 0.05) after day-40 in bred ewes. Secretion of PGF2alpha by day-15 caruncular endometrium of non-bred ewes and bred ewes was increased (P < or = 0.05) by AA on days 13 and 15 and by LH on day-15. Secretion of PGF2alpha by caruncular/placental tissue from bred ewes was (P < or = 0.05) by AA on days 13, 15, 20, 30, and 40 and by LH on days 15, 20, 30, and 40, after which the response decreased (P < or = 0.05). Secretion of PGF2alpha by caruncular endometrium of non-bred ewes during the estrous cycle or by caruncular/placental tissue of bred ewes during the first trimester was not affected by PSPB (P > or = 0.05). Secretion of PGE2 by caruncular endometrium of non-bred ewes did not change (P > or = 0.05) and was increased (P < or = 0.05) by caruncular/placental tissue on days 13-90 from bred ewes when incubated with vehicle. Secretion of PGE2 by endometrium from non-bred ewes was not affected (P > or = 0.05) by AA, LH, or PSPB, but was increased (P < or = 0.05) by AA or LH on days 13-50 and by PSPB on days 60 and 90 when incubated with caruncular/placental tissue from bred ewes. Secretion of progesterone by placental tissue of bred ewes increased (P < or = 0.05) on day-50 and continued to increase through day-90. In summary, uterine/placental tissue secretion of PGF2alpha is not reduced until the end of the first trimester of pregnancy in ewes. In addition, LH appears to play a role in luteolysis of non-bred ewes by stimulating caruncular endometrial secretion of PGF2alpha and on day-5 postbreeding to prevent luteolysis during early pregnancy by stimulating caruncular/placental secretion of PGE2 throughout the first trimester of pregnancy in sheep. Secretion of PGE2 by caruncular/placental tissue after day-50 of pregnancy appears to be regulated by PSPB, not LH. PMID- 12749595 TI - Utilization of physician health care services in Mexico by U.S. Hispanic border residents. AB - One of the most controversial topics in the U.S. is the issue of accessibility to health services by U.S. residents. This issue is most critical to U.S. Hispanic residents living along the U.S.-Mexico border who have been identified as having low health standards and low socio-economic conditions when compared to the rest of the state and the country. The availability of lower cost health services across the U.S. border in Mexico is, therefore, perceived as a viable economic alternative source of health care. This study is derived from a health needs assessment survey of 1,100 households residing in Laredo, Texas, the largest land port along the 2,000-miles long U.S.-Mexico border. The major result of this study indicates that about 41.2 percent of the Laredo U.S. Hispanic residents are utilizing cross border physician health care services in Mexico. PMID- 12749596 TI - Trust in online prescription drug information among internet users: the impact on information search behavior after exposure to direct-to-consumer advertising. AB - The proliferation of both manufacturer-controlled and independent medication related websites has aroused concern among consumers and policy-makers concerning the trustworthiness of Web-based drug information. The authors examine consumers' trust in on-line prescription drug information and its influence on information search behavior. The study design involves a retrospective analysis of data from a 1998 national survey. The findings reveal that trust in drug information from traditional media sources such as television and newspapers transfers to the domain of the Internet. Furthermore, a greater trust in on-line prescription drug information stimulates utilization of the Internet for information search after exposure to prescription drug advertising. PMID- 12749597 TI - Refocusing website marketing: physician-patient relationships. AB - Physician websites have become commonplace in the world of health care marketing. Most, if not all of these websites, focus on practice enhancement tactics as a means of increasing office traffic in an increasingly competitive environment. Websites developed in this way fail to tap the interactive potential of website technology and provide little support for the development of physician-patient relationships. In this paper, we extend a model of medical service care so as to refocus attention on the importance of physician-patient relationships and the role websites can play in this process. PMID- 12749598 TI - Fear, coping, and information: a pilot study on motivating a healthy response. AB - The importance of fear and maladaptive health behavior responses within the ordered protection motivation threat persuasion framework is investigated. By observing the initial creation of maladaptive responses, the results indicate that without adaptive coping response information, high threat information and fear cause avoidance and other maladaptive responses. These influences, in turn, reduce intentions to adopt the recommended adaptive protection behavior. When adaptive coping response information is provided after a threatening fear appeal, maladaptive responses' negative effects are reduced, and adaptive protection behavior intentions are increased. The results of this study have important implications for health and social marketing campaigns (e.g., prevention of AIDS, drug abuse), where maladaptive responses may be responsible for a significant number of program failures. PMID- 12749599 TI - BCG vaccination and tuberculosis in Japan. AB - This paper summarizes Bacillus Calmette-Guerin (BCG) vaccination and revaccination policies in Japan, its cost-effectiveness, side effects, proposed selective vaccination strategy, and present tuberculosis situation in Japanese perspectives based on Medline database and other published reports. Universal BCG vaccination in infants and revaccination among children were not found economically justifiable. Overall tuberculosis incidence in Japan is higher than that of other developed countries. Trend of decline in tuberculosis incidence is similar to that of the countries where universal BCG vaccination has never been implemented. In the recent years, the number of tuberculosis group infection has been escalating. Since BCG revaccination program has already been discontinued, a consensus on universal BCG vaccination is also essential based on social, political, and economical factors. Side by side, more pragmatic strategies such as well-defined tuberculin test, selective vaccination policy based on tuberculosis incidence in each administrative zone, and early vaccination of high risk groups, should be formulated. PMID- 12749600 TI - Annual incidence rate of infectious diseases estimated from sentinel surveillance data in Japan. AB - BACKGROUND: The estimation of incidence rates of infectious diseases based on the sentinel surveillance data is rather rare. We attempted to estimate these in 2000 in Japan by the surveillance data, and to evaluate their biases. METHODS: We used the incidences of influenza-like illness and 12 pediatric diseases in each of the sentinel medical institutions in Japan based on surveillance data in 2000. The incidence in all medical institutions was estimated under the assumption that the sentinel medical institutions were randomly selected. The possible bias of this estimate was evaluated in comparison with the hypothetical true incidence obtained as the total incidence in all medical institutions estimated by a regression model using the numbers of all disease outpatients per day from the National Survey of Medical Care Institutions of Japan. RESULTS: The estimated annual incidence rate was 75.6 (95% confidence interval: 72.3-78.7) per 1,000 population in influenza-like illness, and ranged from 1.1 (95% confidence interval: 1.0-1.2) to 285.2 (95% confidence interval: 270.2-300.3) per 1,000 population aged 0-19 years among 12 pediatric diseases. The ratio of the estimated incidence to the hypothetical true one was 1.06-1.26 among influenza like illness and the 12 pediatric diseases. CONCLUSIONS: The incidence rates of influenza-like illness and pediatric diseases in 2000 in Japan were estimated from sentinel surveillance data. The rates obtained provide some useful but not always accurate information. Thus, further research is necessary. PMID- 12749601 TI - A 6-year cohort study on relationship between functional fitness and impairment of ADL in community-dwelling older persons. AB - We measured functional fitness in older subjects and performed a follow-up survey for 6 years to clarify whether the level of functional fitness at a given point contributes to prediction of the subsequent occurrence of impairment of functions necessary for independent living. The longitudinal data were obtained for 391 persons aged 60 years or over, who were independently living in the community. Four items of functional fitness, i.e. the ability to perform standing/sitting movements, ability to perform traveling movements, ability to perform housekeeping movements, and ability to perform personal grooming activities, were assessed. The relationship between functional fitness and the risk of the occurrence of impairment of independence in daily living was evaluated using a binomial logistic regression model. In males, all the 4 items of functional fitness were significantly related to the risk of impairment of activities of daily living (ADL). In females, however, none of the items was significantly related to the risk of ADL impairment. In conclusion, functional fitness appears to be valid as a predictive parameter of future occurrence of ADL impairment in relatively healthy and independent elderly males. However, its validity in elderly females needs further evaluation. PMID- 12749602 TI - A cohort study on cerebrovascular disease in middle-aged and elderly population in rural areas in Jiangxi Province, China. AB - To clarify the risk factors of CVD deaths in rural areas in Jiangxi Province, China, a cohort study was carried out from September 1, 1994 through December 31, 2000 involving 50,252 participants aged 40 years or older in 4 counties. Among the 3,429 deaths, 671 cases (398 males and 273 females) died of CVD. In addition, excluding 183 cases with a previous history of CVD, 632 CVD deaths out of 50,069 subjects were analyzed using Cox proportional hazard models. The multivariate hazard ratio (HR) for CVD mortality significantly increased in parallel with age, blood pressure and degree of liking for salty foods (p for trend < 0.01). The multivariate HR for CVD mortality of ex-drinkers was 1.55 (95% CI: 1.04, 2.31) compared with non-drinkers. The multivariate HR for CVD mortality of subjects who ate meat once or twice per month was 0.75 (95% CI: 0.62, 0.91) compared with those who never ate meat or seldom. There was no significant relationship between smoking and CVD mortality. Our results indicated that the main risk factors for CVD mortality were advancing age, high-normal blood pressure and hypertension. The risk in these areas was lower in subjects who disliked salty foods and those who ate meat once or twice per month. PMID- 12749603 TI - Prevalence and correlates of hand dermatitis among nurses in a Japanese teaching hospital. AB - BACKGROUND: Although hand dermatitis represents a common occupational disease among hospital nurses, epidemiologic studies of this nature are comparatively rare in Japan. METHODS: We recruited a complete cross-section of nurses from a teaching hospital in central Japan. Data was gathered by means of a self-reported questionnaire, with hand dermatitis symptoms and evaluation criteria drawn from previously validated research. Participants were categorised according to their hospital department during the analysis. RESULTS: A total of 305 questionnaires were successfully completed and returned (response rate: 84%). There were statistically significant differences in hand dermatitis prevalence between the departments (p < 0.05), ranging from 6% in psychiatry to 48% in the surgical unit and averaging 35% across the entire group. A history of allergic disease was shown to increase the risk of hand dermatitis (odds ratio = 3.7, 95% confidence interval: 2.1 - 6.6). Washing their hands more than 15 times per work shift also increased the risk (odds ratio = 2.0, 95% confidence interval: 1.2 - 3.4). CONCLUSION: This study has shown that hand dermatitis prevalence varies among Japanese nurses depending on their hospital department, and is generally quite high when compared to other reports. PMID- 12749605 TI - Analysis of sex, age and disease factors contributing to prolonged life expectancy at birth, in cases of malignant neoplasms in Japan. AB - BACKGROUND: This study aimed to examine the contribution made by the change in mortality from malignant neoplasms to the life expectancy at birth, observed during the years 1965-1995 in Japan. METHODS: We used data on the population and number of deaths by cause, age and sex in 1965, 1975, 1985 and 1995. The contribution of different ages and causes of death to the change in life expectancy were examined with the method developed by Pollard. RESULTS: We found that, among all causes, the decrease of mortality from stomach cancer led to the greatest improvement in life expectancy for both sexes. On the other hand, negative contributions were seen with cancers of many sites, such as cancer of the intestine, liver and lung for males, and cancer of the intestine, gallbladder, lung and breast for females. Recently, the contributing years of all cancers have been negative because of the increase in mortality from malignant neoplasms. In addition, increase of death from malignant neoplasms in middle-aged and elderly people negatively influenced the life expectancy at birth. CONCLUSIONS: Female cancer influenced the improvement in life expectancy at birth. Cancer for males, however, contributed little to improvement of life expectancy at birth except for a little prolongation of life expectancy at birth during the years 1965-1975. To develop a public health policy, the contributing years to life expectancy at birth can be a useful indication in evaluating the impact of death from various diseases. It is necessary to analyze the contribution made by various causes of death to the changes of life expectancy at birth. PMID- 12749604 TI - Effect of diet and Helicobacter pylori infection to the risk of early gastric cancer. AB - BACKGROUND: The association of dietary habits and Helicobacter pylori infection with early gastric cancer is still unclear. METHODS: A hospital-based case control study was conducted in Korea. Sixty-nine patients were newly diagnosed as having early gastric cancer at the Division of Gastroenterology, Asan Medical Center, and 199 healthy subjects who visited the Health Promotion Center of the this same hospital for annual health examinations were selected as controls. Helicobacter pylori infection status was assayed by ELISA, and information for dietary habits was obtained by interview using a semi-quantitative food frequency questionnaires. Preference for salty taste was also evaluated using a sensitive test. RESULTS: H. pylori seropositivity was observed in 88% of cases, as compared with 75% of controls (OR = 5.3, 95% confidence interval:1.7-16.5). Adaptive salt concentration was significantly and positively associated with early gastric cancer risk (p < 0.01). Decreased risks of early gastric cancer were observed in association with intakes of clear broth, raw vegetables, fruits, fruit or vegetable juices, and soybean curds. On the other hand, a high intake of salt fermented fish and kimchi were associated with an elevated risk of early gastric cancer. Subjects with positive H. pylori infection and a high salty preference had a 10-fold higher risk of early gastric cancer than subjects without H. pylori infection and with a low salty preference (p for interaction = 0.047). CONCLUSION: Some dietary factors and H. pylori infection are significantly associated with early gastric cancer. In particular, high-salty diets may enhance the effect of H. pyori infection in gastric carcinogenesis. PMID- 12749606 TI - Association of CYP2A6 gene deletion with cigarette smoking status in Japanese adults. AB - BACKGROUND: Genetic variation of CYP2A6 is shown to alter nicotine metabolism. This study was developed to investigate the genetic influence of the whole deletion-allele of CYP2A6 on active and passive smoking behavior. METHODS: Two hundred and forty Japanese adults, who visited Aichi Cancer Center as outpatients, were genotyped for the wild-type (CYP2A6*1A, CYP2A6*1B) and the whole deletion-type (CYP2A6*4C) polymorphism of CYP2A6. Information about active and passive smoking status was obtained by a self-administered questionnaire. Genetic influence of CYP2A6 polymorphism on smoking behavior was evaluated using the Mantel extension test. RESULTS: The frequency of the deletion allele was 18%. All 8 subjects carrying two deletion alleles had no smoking habit, and the homozygous deletion genotype showed a tendency to correlate with active smoking status after adjustment for sex and age (p = 0.054). However, the proportion of never smokers among heterozygous subjects was almost the same as among subjects carrying no deletion allele (54% and 58%, respectively). Furthermore, CYP2A6 genotypes were correlated neither with the number of cigarettes smoked per day nor with the age at starting smoking (p = 0.364 and 0.880, respectively). Among never smokers, CYP2A6 genotypes were not correlated with exposure to passive smoking at home or in the workplace (p = 0.623 and 0.484, respectively). CONCLUSION: Despite the possible protection against active smoking behavior in subjects homozygous for the deletion allele, the CYP2A6 polymorphism has only a limited impact on public health because no protective effect was found in heterozygous subjects. PMID- 12749607 TI - Hepatitis A vaccination among young African American men who have sex with men in the deep south: psychosocial predictors. AB - Despite recommendations for vaccination against hepatitis A (HAV) of men who have sex with men (MSM), most remain unvaccinated. This study was designed to identify attitudes and beliefs associated with vaccination against HAV using a conventional outreach sample of African American MSM in Birmingham, Alabama. Of 107 participants, nearly 34% reported being vaccinated against HAV. Over half of the participants reported 10 or more different lifetime male sexual partners, and a third reported having had intercourse with females, as well as, males within the past 5 years. About 10% of the participants reported condom use over half of the time during oral intercourse, and 50% of the participants reported using a condom over half the time during anal intercourse. In multivariable analysis, predictors of HAV vaccination were a decreased perception of the practical barriers to HAV vaccination (odds ratio [OR], 0.05; 95% confidence interval [CI]: 0.01-0.18, P = 0.002); increased health provider communication (OR, 9.89; 95% CI: 2.74-35.65, P = 0.02); and increased perceived personal self-efficacy to complete the two-dose series (OR, 7.31; 95% CI: 2.38-22.45, P = 0.02). Our findings underscore the need to increase vaccination through innovative approaches to reduce perceived barriers to vaccination while increasing provider-patient communication and self-efficacy to complete the vaccine series. PMID- 12749608 TI - Increasing immunization rates among African-American adults. PMID- 12749609 TI - Promoting prevention of viral hepatitis in the African American community. PMID- 12749610 TI - Turnaround in thinking on Africa's AIDS crisis. PMID- 12749611 TI - ACOG recommendation for HRT doesn't withstand scientific scrutiny. PMID- 12749612 TI - Depression in medical ill: improving the care. PMID- 12749613 TI - Task force finds evidence lacking on whether routine screening for prostate cancer improves health outcomes. PMID- 12749614 TI - Implications of menopausal hormonal therapy for African-American and Hispanic women. PMID- 12749615 TI - Magnesium deficiency in African-Americans: does it contribute to increased cardiovascular risk factors? AB - African-Americans are known to be disproportionately impacted by many chronic diseases such as diabetes, hypertension, cardiovascular, and renal disease. Lower levels of dietary and serum magnesium have been associated with an increased prevalence of hypertension, insulin resistance, and diabetes. Studies suggest a greater prevalence of occult magnesium deficiency among African-Americans compared to other populations. This increased prevalence of hypomagnesemia may contribute to increased insulin resistance leading to accelerated atherosclerosis and premature death. Trials that correct magnesium status/levels among African Americans, whether through dietary intervention or direct magnesium replacement/supplementation need to be completed to characterize this relationship more completely. PMID- 12749616 TI - Treatment of difficult-to-control blood pressure in a multidisciplinary clinic at a public hospital. AB - Cardiovascular morbidity and mortality in disadvantaged populations remains high. Few innovative strategies or services to treat chronic diseases have been critically analyzed in these patients. We evaluated our initial experiences with a newly established multidisciplinary clinic For the treatment of difficult-to control hypertension and describe reasons for poor blood pressure control as well as treatment strategies. Patients with blood pressures greater than 140/90 despite concurrent treatment with three or more medications for at least three months were referred to our clinic. Data regarding sociodemographic characteristics, health beliefs and behaviors were collected. Two physicians jointly proposed an explanation for lack of blood pressure control. A multidisciplinary team of physicians, nurses, pharmacists, and nutritionists aggressively assessed and reinforced educational objectives tailored to individual needs. 58% of patients achieved target blood pressure at six months, but 22% were lost to follow-up. The most common reasons for previous treatment failure were volume overload and poor medication adherence. We conclude that a multidisciplinary clinic for difficult-to-control blood pressure can be successful in a large, urban hospital serving a disadvantaged minority population. However, more study is needed to delineate the specific reasons for success and further refine treatment strategies. PMID- 12749617 TI - Substance abuse treatment effectiveness of publicly funded clients in Tennessee. AB - The Tennessee Outcomes for Alcohol and Drug Services (TOADS) in collaboration with the Bureau of Alcohol and Drug Abuse Services at the Tennessee Department of Health evaluated the effectiveness of publicly funded substance abuse treatment programs in Tennessee by collecting and analyzing data from clients treated between 1998 and 2000. Using a structured questionnaire, TOADS staff conducted telephone interviews with clients 6 months after their admission to treatment facilities. The sample populations for these follow-up interviews ranged from 1,150 to 1,350 clients over the 3 years, and each year, post-treatment abstinence rates were around 60%, which suggests that treatment in Tennessee has been successful in reducing substance abuse. In addition, the follow-up interview data suggest that treatment also helped drastically reduce both unemployment and arrests among clients. These findings in Tennessee are comparable to treatment outcomes in other states. In addition to the positive effects that treatment has on clients, treatment is also cost-effective for state budgets since treatment reduces many of the burdens substance abuse places on the criminal justice system, the healthcare system, and other state-supported services. PMID- 12749618 TI - A practical approach to fibromyalgia. AB - Fibromyalgia is the name given to a collection of symptoms with no clear physiologic cause, The constellation of symptoms are clearly recognizable as a distinct pathologic entity. The diagnosis is made through clinical observations made by the examiner. Differential diagnosis must include other somatic syndromes as well as disease entities like hepatitis, hypothyroidism, diabetes mellitus, electrolyte imbalance, multiple sclerosis, and cancer. Diagnostic criteria are given as guidelines for the diagnosis, not as absolute requirements. Treatment of this condition remains individualized and relies heavily on having a therapeutic relationship with a provider. Treatment of this syndrome needs to be looked at as an ongoing process. Goal oriented treatment aimed at maintaining specific functions can be directed at helping a patient get restorative sleep, alleviating the somatic pains that ail the patient, keeping a person productive, regulating schedules or through goal oriented agreements made with the patient. Since this syndrome is chronic and may effect all areas of a persons functioning the family and social support system of the person being treated need to be evaluated. Patients often seek alternative medical treatments for this problem including diet therapy, acupuncture, and herbal therapy. Treatment must involve more than just the symptoms presented and the patient can only be treated successfully if they are willing to work at changing their own perceptions, and ways of relating to stressors in their world. PMID- 12749620 TI - SNMA's presidential initiative: mental health and minority communities. PMID- 12749619 TI - Immediate breast reconstruction-impact on radiation management. AB - Breast reconstruction is an option for women undergoing modified radical mastectomy due to a diagnosis of breast cancer. In certain patients, breast reconstruction is performed by insertion of a temporary tissue expander prior to the placement of permanent breast implants. Some of these patients, following mastectomy, may require chest wall irradiation to prevent loco regional relapse. The compatibility of radiation and tissue expanders placed in the chest wall is of major concern to the radiation oncologist. Clinically undetectable changes can occur in the tissue expander during the course of radiation therapy. This can lead to radiation treatment set-up changes, variation in tissue expansion resulting in unwanted cosmesis, and deviation from the prescribed radiation dose leading to over and/or under dosing of tumor burden. At Howard University hospital, a CT scan was utilized to evaluate the status of the temporary tissue expander during radiation treatment to enable us to prevent radiation treatment related complications resulting from dosimetric discrepancies. CT images of the tissue expander were obtained through the course of treatment. To avoid a 'geographic miss' the amount of fluid injected into the tissue expander was kept constant following patient's satisfaction with the size of the breast mound. The CT scans allowed better visualization of the prosthesis and its relation to the surrounding tumor bed. This technique ensured that anatomical changes occurring during radiation treatment, if any, were minimized. Repeated dosimetry evaluations showed no changes to the prescribed dose distribution. A CT of the reconstructed breast provides an important quality control. Further studies with greater number of patients are required for confirming this impact on radiation treatment. PMID- 12749621 TI - Scheuermann's disease as a model displaying the mechanism of venous obstruction in thoracic outlet syndrome and migraine patients: MRI and MRA. AB - Kyphosis of the thoracic spine rotates the scapulae anterior laterally, clavicles and subclavius muscles anteriorly, displaces the manubrium posteriorly, which increases the slope of the first ribs. This increases tension on the anterior scalene muscles and the neurovascular bundles which causes brachial plexopathy (TOS). Scheuermann's disease (spinal osteochondrosis; juvenile kyphoscoliosis) is a disorder which consists of vertebral wedging endplate irregularity and narrowing of the intervertebral disk space causing kyphosis of the thoracic spine and may also involve the lumbar space. It occurs at puberty and involves both male and females. Abduction external rotation of the upper extremities (arms overhead) posterior inferiorly rotate the clavicles and the subclavius muscles which enhances tension on the venous drainage and neurovascular supply that diminishes venous return. This triggers complaints of thoracic outlet syndrome (TOS) and migraine headache. Bilateral magnetic resonance imaging (MRI) demonstrates compressing abnormalities of the brachial plexus. Five patients with Scheuermann's disease were imaged with the 1.5 Tesla magnet (Signa; General Electric Medical Systems, Milwaukee, WI) 3-D reconstruction MRI. T1W and T2W pulse sequences were performed in transverse, the coronal, transverse oblique, sagittal, and coronal abduction external rotation planes using 4 mm slice thickness and 512 x 256 matrix size. Water bags were used to enhance the signal to noise ratio. Magnetic resonance angiography (MRA) 2-D Time Of Flight (TOF) was obtained to compression for anatomic display evaluate perfusion of the brachial plexus. MRI and MRA captured sites of brachial plexus. One patient was selected for this presentation, which demonstrates the compression of the brachial plexus and venous obstruction which triggered complaints of thoracic outlet syndrome. PMID- 12749622 TI - Race and medical publications. PMID- 12749623 TI - Human genome, race and medicine. AB - In closing, with the new revelations of the Human Genome Project, notions on whether land-based race identity or ethnicity with genetic markers has been proven valid, with few exceptions. This has caused me to revisit the attempted effort to discard concepts on race, especially in medicine. Obviously there are outliers to this new work. But contrary to popular belief, and to some, the unthinkable, there may be, in fact, a biologic basis for our human distinctions. And I for one do not feel shame or seem perplexed by it. Moreover, it appears that Dr. Welsing in her earlier work was onto something, and was indeed ahead of her time. The problem African Americans, and other persons of color face, to some extent, has to do with the social political context of racism, and the biologic impact it has and is often expressed in the form of stress and injuries, simply put. Therefore, and more importantly, eliminating the nomenclature of how we classify ourselves in our intellectual interchange in science and other areas, will not correct our problems, but may in fact, if abandon, spell our doom. Because what we are murderously burdened by has to do with racism and its effects. Which are in effect, based on physical features, not mere classification. Further, the current thought on racism and why it is practiced by some is that racism serves an evolutionary drive to survive by humans, by forming alliances in among similar groups of people. Therefore, if that is the case, we had better be ready for the long haul in this battle as our history and ongoing struggles tell us. Besides, if not for racism, "we would not have had all of these problems over all these years." The National Medical Association and its publishing instruments must remain vigilant and stay focused. PMID- 12749625 TI - Human rights, environment, and individual action. PMID- 12749624 TI - Unrealistic expectations of pediatricians and smoking cessation counseling. PMID- 12749626 TI - Rights and reality: monitoring the public's right to participate. AB - Exercising the right to a sustainable and healthy environment requires the ability to participate in government and business decisions that affect the environment. A global coalition of public interest groups and research organizations measured the public's ability to participate in such decisions in nine countries representing a range of income levels and development paths. The investigation assessed the three elements of public participation as defined at the Rio Earth Summit in 1992: access to information, access to the decision making process, and access to redress. The results show that, despite considerable progress in these areas, serious gaps remain in the public's ability to participate in important environmental decisions. These gaps occur in both developed and developing nations, and reflect limited articulation of participation rights in national law and institutional practice. The results also suggest that the public is often unfamiliar with and unaccustomed to exercising the rights they already have. PMID- 12749627 TI - Toxic disputes and the rise of environmental justice in Australia. AB - The paper examines the rise of environmental justice issues in Australia, evident in two toxic disputes; the first, in a Perth outer suburb in Western Australia where residents faced both a hazardous waste dump and the nation's biggest chemical fire; and the second, in the Sydney suburb of Botany where residents were confronted with the destruction of what is thought to be, the world's largest stockpile of hazardous hexachlorobenzene (HCB) waste. The paper reviews the range of factors that impacted the local communities' fight for environmental justice. It explores the limitations of risk assessment and risk-based policies, as well as the problematic role of the expert and the communication of risk. The informational inequity and resource disparities so evident in toxic disputes are highlighted. The case studies confirmed the inequitable distribution of chemical risk as a failure to secure environmental justice for all Australians. PMID- 12749628 TI - Fluoridation: a violation of medical ethics and human rights. AB - Silicofluorides, widely used in water fluoridation, are unlicensed medicinal substances, administered to large populations without informed consent or supervision by a qualified medical practitioner. Fluoridation fails the test of reliability and specificity, and, lacking toxicity testing of silicofluorides, constitutes unlawful medical research. It is banned in most of Europe; European Union human rights legislation makes it illegal. Silicofluorides have never been submitted to the U.S. FDA for approval as medicines. The ethical validity of fluoridation policy does not stand up to scrutiny relative to the Nuremberg Code and other codes of medical ethics, including the Council of Europe's Biomedical Convention of 1999. The police power of the State has been used in the United States to override health concerns, with the support of the courts, which have given deference to health authorities. PMID- 12749629 TI - Greater risks, fewer rights: U.S. farmworkers and pesticides. AB - Pesticide Action Network, United Farmworkers of America, and California Rural Legal Assistance Foundation analyzed California government data on agricultural poisonings and enforcement of worker safety standards. Nearly 500 pesticide poisonings were reported for California farmworkers every year from 1997 to 2000. The actual number of pesticide-related illnesses is unknown, since many poisonings go unreported. Most poisonings occurred as a result of soil fumigation and pesticide applications to grapes, oranges, and cotton. Pesticide drift accounted for 51% of the cases, and another 25% resulted from exposures to pesticide residues. Violations of worker safety laws were common, contributing to 41% of reported poisonings. No violations occurred in another 38%, indicating that existing laws inadequately protect workers from pesticide exposure. This snapshot of human rights abuse through pesticide exposure in California-the site of some of the world's most stringent pesticide use and worker safety laws illustrates the global problem of pesticide poisoning among agricultural workers. PMID- 12749630 TI - Pesticides and human rights. AB - Many nations, including those in developing countries, whose workers are the most susceptible to exploitation, have in place statutes and regulations to protect the rights to life, health, and livelihoods of their people. Some of these efforts to protect human rights are detailed, and specific instances in which the protections they were intended to offer have been ineffective are described. The authors argue for adding strength to States' legislation to truly protect human rights. PMID- 12749631 TI - The tragedy of Tauccamarca: a human rights perspective on the pesticide poisoning deaths of 4 children in the Peruvian Andes. AB - The pesticide poisoning deaths of 24 children in an isolated Peruvian village make a compelling case that corporate accountability for pesticide poisonings in the developing south should be examined from a human rights perspective. Highly toxic pesticides cannot be used safely under prevailing socioeconomic conditions. The industry asserts that the deaths of these children were accidental, blaming misuse. Tragedies such as these poisonings are not accidents, but foreseeable, and therefore preventable. Sales of highly toxic pesticides that cause repeated and predictable poisonings violate the fundamental human rights to life, health, and security of person. The Tauccamarca tragedy is a clear example of the urgency of applying a precautionary, human rights approach to pesticide issues in the developing south. PMID- 12749632 TI - Human rights, environmental justice, and the health of farm workers in South Africa. AB - Despite the democratization of South Africa in 1994, which brought the agricultural sector within the ambit of legal protection, farm workers remain vulnerable to an undue burden of social and health problems. Alcohol abuse due to the DOP system, pesticide poisonings, and other occupational hazards illustrate that the likely success of efforts at redress depends on a greater awareness of the rights and justice dimensions of the health problems facing these workers. International trade policies may exacerbate inequalities that deprive them of opportunities to realize their rights at national level. A public health agenda must integrate into programs and policies to address the health of farm workers the recognition that violations of their rights underlie much of their burden of ill health. PMID- 12749633 TI - Human rights: necessary? Sufficient? Diversionary? AB - Claims of human rights have historically been a response to the violence and oppression brought by some people onto others. Focusing on the individual person, modern concepts of human rights inadequately address the relationships of individual people to their communities and rarely address relationships of people with other species and ecological systems more generally. During the past 50-100 years the world has undergone profound ecological change, and although concepts of human rights remain useful, their limits are becoming increasingly clear. Advances in ecological and biological sciences demonstrate dialectic relationships among components and the whole of complex systems. Where the individual begins and ends is unclear. A new ethic that incorporates new ecological understanding is essential in order to address the essentially new world of today. A deep sense of responsibility and an ethic of care, trust, respect, and reciprocity are essential to this undertaking. PMID- 12749634 TI - Accountability in the pesticide industry. AB - To counter the lack of corporate accountability of the agrochemical industry for the damage caused by its perpetuation of the use of harmful chemical pesticides, the Rockefeller Brothers Fund staff in June 2002 brought together concerned scientists, lawyers, socially responsible investment professionals, and sustainable agriculture advocates at their Pocantico Conference Center. The group's objective was to communicate to market analysts the long-term downside risks of investments in pesticides, in the hope that dissemination of this information would contribute to increasing corporate accountability and safeguarding public and environmental health. Excerpts from its proceedings are presented. PMID- 12749635 TI - Integrity of the conduct of the IARC monographs program. PMID- 12749636 TI - Reply to Dr. Sass. PMID- 12749639 TI - Suppression of crucial information in the IARC evaluation of DEHP. PMID- 12749640 TI - Comment from the union participant in the IARC Working Group that downgraded DEHP. PMID- 12749641 TI - Continued insensitivity to conflicts of interest at IARC. PMID- 12749642 TI - The IARC's role in public health. PMID- 12749643 TI - WHO handling of conflicts of interest. PMID- 12749644 TI - The Bhopal disaster: prevention should have priority now. PMID- 12749645 TI - Economic costs of care in extremely low birthweight infants during the first 2 years of life. AB - OBJECTIVE: To assess the 2-yr costs of extremely low birthweight infants' (ELBWIs; birthweight, < 1000 g) care in relation to birthweight, outcome, and the costs of normal birthweight infants. DESIGN: Cost data were obtained from care giving hospitals and by a parental questionnaire. Outcome data from the perinatal and neonatal periods and from the first 2 yrs for both ELBWIs and control infants had been prospectively collected to a national ELBWI register. PATIENTS: We studied 71 ELBWIs and 60 normal birthweight controls born in Helsinki University Hospital in 1996-1997. MEASUREMENTS: Collected data comprised costs resulting from care of ELBWIs and normal birthweight control infants and included hospital, outpatient care, medication, rehabilitation, auxiliary means, and travel costs; ancillary costs from daily care; parent's accommodation during hospitalization periods; and loss of earnings during the infant's first 2 yrs. MAIN RESULTS: The average total 2-yr healthcare cost was 104,635 Euros for surviving ELBWIs and 3,135 Euros for control infants. In ELBWIs, initial hospital costs alone accounted for 64% of total costs; the costs during the first and second postdischarge years accounted for 20% and 13%, respectively. The mean hospital cost of nonsurviving ELBWIs was 19,950 Euros. A normally developed ELBWI had costs 25-fold, a mildly disabled ELBWI had costs 33-fold, and a severely disabled ELBWI had costs 68-fold those of control infants. Birthweight correlated negatively with intensive care costs but did not correlate with costs after initial discharge. CONCLUSION: Total costs of ELBWIs decreased over time up to the age of 2 yrs, but even in normally developed ELBWIs, costs remained higher than those of normal birthweight infants. Low birthweight seemed to be related to increased initial hospital costs but not to annual costs after the first discharge. PMID- 12749646 TI - Dying in the intensive care unit: collaborative multicenter study about forgoing life-sustaining treatment in Argentine pediatric intensive care units. AB - OBJECTIVE: Describe modes of death and factors involved in decision-making together with life support limitation (LSL) procedures. DESIGN: Prospective, descriptive, longitudinal, and noninterventional study. SETTING: Sixteen pediatric intensive care units in Argentina. PATIENTS: Every patient who died during a 1-yr period was included. MEASUREMENTS AND MAIN RESULTS: Age, sex, length of stay (LOS), primary and admission diagnosis, underlying chronic disease (CD), postoperative condition (PO). Deaths were classified in four groups: a) failed cardiopulmonary resuscitation (CPR); b) do-not-resuscitate (DNR) status; c) withholding or withdrawing life-sustaining treatment (WH/WD); and d) brain death (BD). Justifications were classified as a) imminent death; b) poor long term prognosis; c) poor quality of life; and d) family request. Data were collected from medical records and interviews with the attending physicians. Descriptive statistics were performed. Differences among groups were analyzed through contingency tables and analysis of variance when required. Relative risks and confidence intervals of variables potentially related to LSL were analyzed, and logistic regression was performed. There were 6358 admissions and 457 deaths. CPR was performed in 52%, DNR in 16%, WH/WD in 20%, and BD in 11% of dead patients. BD patients were older, LOS and CD prevalence were higher in the WH/WD group. Inotropic drugs were the most frequently limited treatment in 110 patients (55%), CPR in 72 (35.6%), and mechanical ventilation in 63 (31%). Imminent death was the most frequently reported justification for LSL. CD and more staff were associated with a higher probability of LSL. CONCLUSIONS: Most of the patients in Argentina underwent CPR before their death. We have a high proportion of patients with CD (65%) and low BD diagnosis. PO condition decreased LSL probability in chronically ill patients. Do-not-resuscitate orders and withholding new treatments were the most common LSL. Active withdrawal was exceptional. The Ethics Committee was consulted in 5% of the LSL population. PMID- 12749647 TI - Elevated cerebrospinal fluid levels of glutamate in children with bacterial meningitis as a predictor of the development of seizures or other adverse outcomes. AB - OBJECTIVE: Evaluation of elevated cerebrospinal fluid levels of glutamate in children with bacterial meningitis as a predictor of seizures or other adverse outcomes. DESIGN: Prospective cohort study with controls. SETTING: A 36-bed pediatric intensive care unit and primary pediatric referral center. PATIENTS: From 1999 to 2001, a total of 55 patients, between the ages of 0 and 18 yrs, with lumbar punctures performed for suspected meningitis. MEASUREMENTS AND MAIN RESULTS: A total of 23 patients had bacterial meningitis confirmed by cerebrospinal fluid/blood culture and elevated cerebrospinal fluid white blood cell counts, and 32 patients, who tested negative, were included as controls. The median age for the patients with meningitis was 1.0 yr (range, 0.0-15.2 yrs), and in the culture-negative group (control group), the median age was 0.3 yrs (range, 0.0-17.0 yrs). The average cerebrospinal fluid white blood cell count was 2707 +/ 3897 in the group with bacterial infection, whereas in the control group, the average was 148 +/- 259 (p < .01). Patients with bacterial meningitis had a mean cerebrospinal fluid glutamate level of 60.5 +/- 88.4 mol/L, whereas the mean cerebrospinal fluid glutamate level in the control group was 4.9 +/- 11.0 mol/L (p < .01). However, only 10 of 23 children with bacterial meningitis had a second lumbar puncture performed during the study. There was no correlation between the cerebrospinal fluid white blood cell count and cerebrospinal fluid glutamate levels in either the study or control patients. None of the control patients developed seizures or neurologic deficits, despite some patients having elevated glutamate levels. However, four patients with bacterial meningitis developed seizures after admission to the hospital, and ten were discharged with at least some neurologic sequelae attributable to their infection. Two out of the three who developed seizures and had a repeat lumbar puncture demonstrated persistent elevation of cerebrospinal fluid glutamate levels. In addition, 70% of patients (7 of 10) with Streptococcus pneumoniae meningitis developed neurologic complications (p = .04). CONCLUSIONS: Bacterial meningitis in children causes an increase in cerebrospinal fluid glutamate that in many cases persists over time. However, in this limited study, neither higher nor persistent elevation of cerebrospinal fluid glutamate levels is predictive of which patients might develop seizures or other apparent immediate adverse outcomes after invasive infection. The responsible organism seems to have far more significance in predicting the development of adverse sequelae. PMID- 12749648 TI - Can energy expenditure be predicted in critically ill children?. AB - OBJECTIVE: To determine whether critically ill children are hypermetabolic and to calculate whether predictive equations are appropriate for critically ill children. DESIGN: Prospective, clinical study. SETTING: Pediatric intensive care unit. PATIENTS: A total of 57 children (39 boys) aged 9 months to 15.8 yrs. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The median resting energy expenditure measurement measured by indirect calorimetry was 37.2 (range, 11.9 66.6) kcal x kg(-1) x day(-1). This was significantly lower than would be predicted using either the Schofield (42.7 [26.9-65.4] kcal x kg(-1) x day(-1)) or Fleisch equations (42.8 [20.9-66.2] kcalx kg(-1)-1 x day(-1), p < .001) but significantly higher than the White equation developed specifically for pediatric intensive care units (26.2 [8.5-70.1] kcal x kg(-1),day(-1), p < .0001). Methods comparison analysis showed the limits of agreement were -484 to 300, -461 to 319, and -3.2 to 854 kcal/day, respectively. Multivariate analysis indicated the following factors contribute to hypometabolism and hypermetabolism: age (p = .006), sex (p = .034), time spent in the pediatric intensive care unit (p = .001), diagnosis (p = .015), weight (p = .009), temperature (p = .04), continuous infusion for sedation (p = .04), and neuromuscular blockade (p = .03). CONCLUSION: Children do not become hypermetabolic during critical illness. These data suggest that agreement between resting energy expenditure and the predictive equations are so broad that they are inappropriate for use in critically ill children. PMID- 12749649 TI - Multiple organ dysfunction syndrome in children. AB - OBJECTIVE: Delineation of multiple organ dysfunction syndrome (MODS) is important because of its frequent occurrence in the pediatric intensive care unit and its association with high mortality. However, studies in children are scarce, all have been done in developed countries, and, unlike adult studies, some showed that sepsis is not related to mortality. The aim of this study was to learn about the epidemiology of MODS in our pediatric intensive care unit and to observe if sepsis is associated with mortality. DESIGN: Prospective, observational study. SETTING: A 16-bed pediatric intensive care unit. PATIENTS: A total of 269 patients admitted to the pediatric intensive care unit during the study period from August 1996 to January 1997. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: There were 276 admissions (269 patients) during the study period. A total of 156 of them (56.5%) had MODS at some time during their pediatric intensive care unit stay, and 132 (84.6%) of the children had it at the time of admission. There were 71 deaths during the study period, and 65 of them (91.5%) had MODS. The organs less frequently involved were gastrointestinal and hepatic. In children with MODS, 87 had sepsis, and mortality in this group (51.7%) was greater than in those who did not present with sepsis (28.9%, p < .001). A Pediatric Risk of Mortality score of > or =15 and the presence of sepsis were associated with an increase in mortality risk (odds ratio, 2.8; 95% confidence interval, 1.36-5.75; and odds ratio, 2.33; 95% confidence interval, 1.18-4.59; respectively). CONCLUSIONS: MODS in children usually occurs early, and sepsis increases mortality. Hepatic and gastrointestinal failures are infrequent, and as has been suggested, they could be excluded from the majority of MODS diagnoses. PMID- 12749650 TI - Cephalad movement of endotracheal tubes caused by prone positioning pediatric patients with acute respiratory distress syndrome. AB - OBJECTIVE: To test the hypothesis that prone positioning of patients with acute respiratory distress syndrome results in significant cephalad movement of their endotracheal tubes (ETT). DESIGN: A retrospective review of chest radiographs and patient information. SETTING: Pediatric intensive care unit of a children's hospital. MEASUREMENTS AND MAIN RESULTS: Patients with acute respiratory distress syndrome had digital chest radiographs performed before and immediately after prone positioning as per our routine practice. Based on measurements of the length of the thoracic trachea and the length of the thoracic segment of the ETT, the movement of the ETT subsequent to prone positioning was calculated. Fifteen pairs of radiographs of 14 consecutive patients were evaluated. There were seven girls and seven boys, with ages ranging from 2 months to 18 yrs. All patients had a cephalad movement of their ETT ranging from 10% to 57% of their thoracic tracheal length (p < .001) associated with prone positioning. The mean amplitude of this movement was 34% +/- 16%, indicating that if the tip of the ETT is not deeper than one third of the thoracic tracheal length before prone positioning, it might slide into the cervical trachea as a result of this procedure. CONCLUSIONS: Prone positioning results in cephalad movement of ETT within the trachea. The tip of the ETT should be deeper than one third of the total length of the thoracic trachea before prone positioning to prevent it from moving into the cervical trachea. When prone positioning is done with an ETT originally not deeper than one third of the thoracic trachea, obtaining a chest radiograph immediately after prone positioning is important to determine whether the ETT remained safely situated in the trachea. PMID- 12749652 TI - Pediatric critical care medicine: planning for our research future. AB - OBJECTIVE: To introduce to the pediatric critical care medicine community a new program in pediatric critical care medicine at the National Institutes of Health. DATA SOURCE: Summary of literature review and conference proceedings. DATA SYNTHESIS: At the National Institute of Child Health and Human Development (NICHD), a new program in pediatric critical care and rehabilitation research has been established in the National Center for Medical Rehabilitation Research. The program is directed by a pediatric intensivist and is focused on developing research directed toward improving long-term outcomes in pediatric critical care and on incorporating pediatric rehabilitation medicine as a partner in this goal. To provide strategic direction for the new program, the NICHD sponsored a planning conference May 3-4, 2002, at the NICHD in Bethesda, MD. The conference invitees represented a broad range of pediatric critical care medicine clinical and research interests, expertise, and career stages. It also included individuals with expertise in rehabilitation research. CONCLUSION: The composition of the new program, including its link to physical medicine and rehabilitation, is discussed. In addition, recommendations by the conference participants and program director are provided to foster the development of more randomized, controlled clinical trials and to develop successful clinician scientists in pediatric critical care medicine. PMID- 12749651 TI - Serum procalcitonin in children with suspected sepsis: a comparison with C reactive protein and neutrophil count. AB - OBJECTIVES: To investigate the specific characteristics of serum procalcitonin in children with severe infection, to identify relevant factors influencing procalcitonin increase, to assess its prognostic value, and to compare it with C reactive protein and neutrophil count DESIGN: A prospective observational study and 48 hrs of follow-up of a cohort of cases. SETTING: A pediatric intensive care unit within a children's university hospital in collaboration with a laboratory mainly involved in research in pediatric clinical immunology. PATIENTS: A total of 80 children (median age, 3.1 yrs; range, 1 month to 16 yrs) admitted to a pediatric intensive care unit by suspicion of sepsis. INTERVENTIONS: All patients were treated according to a protocol using antibiotics, fluid resuscitation, inotropic drugs, and mechanical ventilation when they presented with shock or respiratory failure. MEASUREMENTS AND MAIN RESULTS: Serum procalcitonin and C reactive protein were measured at admission in all patients and, when possible, repeated 6,12, 24, and 48 hrs later. In most cases, serum procalcitonin was already very high at onset (range, 1.0-722 ng/mL), and it did not increase significantly afterward. Contrary to C-reactive protein, serum procalcitonin did not vary according to the age of patients. The increase of procalcitonin was higher in patients with shock or multiple organ dysfunction syndrome, having a high severity score (Pediatric Risk of Mortality) or in patients who later died. CONCLUSIONS: Serum procalcitonin levels show a rapid increase in children with sepsis, even in infants < 12 month old, and they have a better prognostic value than C-reactive protein or neutrophil count. PMID- 12749653 TI - Development of bradycardia during sedation with dexmedetomidine in an infant concurrently receiving digoxin. AB - OBJECTIVE: To describe the development of bradycardia during sedation with dexmedetomidine in a patient concurrently receiving digoxin. DESIGN: Case report. SETTING: The pediatric intensive care unit of a tertiary care children's hospital. PATIENTS: A 5-wk-old infant with an atrioventricular septal defect requiring sedation during mechanical ventilation for acute respiratory syncytial virus infection. MEASUREMENTS AND MAIN RESULTS: As part of an ongoing evaluation of dexmedetomidine for sedation in the pediatric intensive care unit, the patient received a loading dose (0.5 microg/kg) followed by an infusion (0.44 microg x kg(-1) x hr(-1)) of dexmedetomidine. Sedation assessments and hemodynamic data were collected at least every 2 hrs. During the loading dose, the patient's heart rate decreased from 133 beats/min per min to 116 beats/min. During the ensuing 13 hrs, the heart rate continued to decrease into the mid 90s, with additional episodes of bradycardia into the 40s and 50s. Within 1 hr of discontinuation of the dexmedetomidine infusion, the baseline heart rate had recovered, and no further episodes of acute bradycardia were noted. CONCLUSIONS: This case adds to the limited data regarding dexmedetomidine in pediatric critical care and suggests that caution should be used when considering sedation with dexmedetomidine in patients also receiving digoxin. PMID- 12749654 TI - Mechanically ventilated pediatric stem cell transplant recipients: effect of cord blood transplant and organ dysfunction on outcome. AB - OBJECTIVES: To compare survival of pediatric umbilical cord blood and bone marrow transplant recipients requiring admission to a pediatric intensive care unit for mechanical ventilation and to determine the effect of organ dysfunction on outcome. DESIGN: Retrospective chart review. SETTING: Tertiary care referral center for pediatric stem cell transplants. PATIENTS: All children 0-18 yrs old admitted to the pediatric intensive care unit for mechanical ventilation after receiving a stem cell transplant. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Data were collected from medical records of 86 patients who received a stem cell transplant and were subsequently admitted to the pediatric intensive care unit for mechanical ventilation. Demographic data were collected at the time of intubation, and physiologic data were collected at 6 hrs and 96 hrs after intubation. The pediatric intensive care unit, hospital, and 2-yr survival rates for umbilical cord blood transplant recipients were 37%, 25%, and 19%, respectively. The survival rates for bone marrow transplant recipients were 47%, 32%, and 21% for the same time periods. Umbilical cord blood and bone marrow transplant recipients with hepatic dysfunction had a significantly worse outcome, as did patients admitted for respiratory failure or sepsis. CONCLUSIONS: Pediatric recipients of an umbilical cord blood transplant who subsequently required mechanical ventilation had lower pediatric intensive care unit and hospital survival rates compared with patients receiving bone marrow transplantation. Survival at 2 yrs for umbilical cord blood transplant and bone marrow transplant patients was similar. Predictors of outcome for all stem cell transplant recipients requiring mechanical ventilation included pediatric intensive care unit diagnosis requiring intubation and hepatic function. Predictors of outcome can be identified shortly after intubation in pediatric stem cell transplant recipients and may aid in therapeutic decision making and family counseling. PMID- 12749655 TI - Parental experience of highly technical therapy: survivors and nonsurvivors of extracorporeal membrane oxygenation support. AB - OBJECTIVE: To describe the experience of parents of critically ill infants and children who require highly technical therapy and to identify interventions that parents might find helpful during the experience. DESIGN: Nonexperimental 8-yr descriptive study. SETTING: Pediatric intensive care unit within a children's hospital. PATIENTS: Parents of infants and children who were supported on extracorporeal membrane oxygenation (ECMO). INTERVENTIONS: Questionnaires, mailed 4-months after ECMO support, inquired about parent preparation and emotional responses, concerns, family-staff communication related to ECMO, and whether, retrospectively, parents would have consented again to ECMO. MEASUREMENTS AND MAIN RESULTS: Two hundred seventy-two parents responded. More than 25% of parents reported being reassured by the hope of therapy. Sixty percent reported they felt they really did not have a choice but to consent to ECMO, given the severity of their child's condition. Upon first seeing their child on ECMO support, most parents reported feeling relieved to see their child alive and being as well prepared as possible. Although some parents remained anxious throughout the ECMO course, talking with care providers and observing the child's daily progress lessened parental fears. Although parents in the nonsurvivor group recalled discussing death before ECMO, 22% remembered first hearing about the possibility of death only after their child failed to improve while undergoing ECMO. During the process of ECMO withdrawal, nearly one fourth of the nonsurvivor group expected their child to fool everyone and to live. After their child's death, 93% felt comforted by follow-up contact with hospital personnel. Compared with the survivor group, fewer parents of nonsurvivors reported that they would again consent to ECMO. CONCLUSIONS: Researched-based family-driven care requires an understanding of parental experience and worries, and interventions that parents themselves identify as helpful. Continued sensitive individualized care is warranted. PMID- 12749656 TI - Outbreak of invasive disease caused by methicillin-resistant Staphylococcus aureus in neonates and prevalence in the neonatal intensive care unit. AB - OBJECTIVES: To describe an outbreak of severe invasive disease caused by methicillin-resistant Staphylococcus aureus (MRSA) and the epidemiology of MRSA in a neonatal intensive care unit during a 12-yr period from 1989 to 2001. SETTING: A 40-bed, level III neonatal intensive care unit at a children's hospital that admits approximately 450 neonates each year from about 35 neighboring hospitals. PATIENTS: All neonates infected or colonized with MRSA during the outbreak are described. All cases of MRSA infection or colonization in the neonatal intensive care unit from 1989 to 2001 were identified from the database maintained by the hospital epidemiology program. RESULTS: During the outbreak, 12 neonates were infected or colonized with MRSA, 11 of whom had the epidemic strain. Seven of these 11 neonates had invasive disease, including bacteremia, meningitis, or urinary tract infection, and four neonates were colonized with the epidemic strain. This outbreak was difficult to control by routine epidemiologic measures, and additional control measures, including closing the neonatal intensive care unit to new admissions and treating all infants with intranasal mupirocin, were implemented. Since the outbreak, the prevalence of MRSA in the neonatal intensive care unit has remained low. CONCLUSIONS: MRSA outbreaks in neonatal intensive care units can be prolonged. Aggressive infection-control measures are often necessary to terminate these outbreaks. Such efforts are essential because MRSA infections in premature neonates can cause significant morbidity and mortality. PMID- 12749657 TI - Correction factors for oxygen and flow-rate effects on neonatal Fleisch and Lilly pneumotachometers. AB - OBJECTIVE: To assess the effects of different oxygen concentrations and flow rates on the measurement errors of neonatal pneumotachometers in heated and unheated situations and to develop correction factors to correct for these effects. DESIGN: Prospective laboratory study. SETTING: Outpatient clinic with equipment in a standardized setting. SUBJECTS: Neonatal pneumotachometers. INTERVENTIONS: In standardized conditions, the tested pneumotachometer was calibrated at a flow rate of 3 L/min with 60% oxygen and was set in series with a closed spirometer system being used as a reference. Different air-flow levels (1 9 L/min) and oxygen concentrations (21-100%) were infused into the closed system with the pneumotachometer and spirometer. MEASUREMENTS AND MAIN RESULTS: The pneumotachometers were significantly affected by changing oxygen concentrations (p < .01) and increasing flow rates (p < .01), increasing the actually measured flow rate. Correction factors, developed by multiple regression analysis, significantly reduced the overall maximum errors of the pneumotachometers from 1.1 to 0.6 L/min to -0.5 to 0.4 L/min. CONCLUSIONS: The effects of changes in oxygen concentrations and flow rates on neonatal pneumotachometers could be considerably decreased by the use of correction factors such as were calculated in this study. This will preclude frequent calibration procedures with actual flow and oxygen levels during changes in experimental settings. PMID- 12749658 TI - Perfluorooctyl bromide (perflubron) attenuates oxidative injury to biological and nonbiological systems. AB - OBJECTIVE: To examine whether perfluorooctyl bromide (perflubron) is capable of protecting biological and nonbiological systems against oxidative damage through a mechanism independent of its known anti-inflammatory property. DESIGN: A controlled, in vitro laboratory study. SETTING: Research laboratory of a health sciences university. SUBJECTS: Rat pulmonary artery endothelial cell cultures (biological system) and linoleic acid in sodium dodecyl sulfate micelles (nonbiological system). INTERVENTIONS: Rat pulmonary artery endothelial cells labeled with dichlorofluorescein diacetate and incubated with perflubron or culture media (control) were exposed to H2O2. H2O2-induced fluorescence of dichlorofluorescein diacetate was measured as an index of intracellular oxidative stress. In another experiment, linoleic acid in sodium dodecyl sulfate micelles was exposed to various concentrations of the azo initiator 2,2'-diazo-bis-(2 amidinopropane) dihydrochloride (2, 4, 20, and 50 mM) in the presence or absence of perflubron. Malondialdehyde measurements were obtained as a marker of oxidative damage to linoleic acid. MEASUREMENTS AND MAIN RESULTS: Cell monolayers incubated with perflubron exhibited 66.6% attenuation in intracellular fluorescence compared with controls (p < .05). Linoleic acid in sodium dodecyl sulfate micelles incubated with perflubron and exposed to 2, 4, 20, or 50 mM of 2,2'-diazo-bis-(2-amidinopropane) dihydrochloride showed less evidence of lipid peroxidation as indicated by lower malondialdehyde measurements at 240 mins (10.6%, 16%, 41%, and 14.2%, respectively) compared with controls. CONCLUSIONS: Perflubron attenuates oxidative damage to both biological and nonbiological systems. This newly recognized property of perflubron is independent of its anti inflammatory properties. PMID- 12749659 TI - Use of hypertonic saline for the treatment of altered mental status associated with diabetic ketoacidosis. PMID- 12749661 TI - Acute subdural hematoma after caesarean section: a case report. AB - OBJECTIVE: The objective of this article is to describe a case of acute subdural hematoma in an infant after difficult delivery and to describe the literature regarding therapy and outcomes of these patients. DESIGN: Case report. PATIENT: Critically ill male newborn who suffered from acute subdural hematoma after a difficult caesarean section. RESULTS: A craniotomy and evacuation of the acute subdural hematoma was performed. At discharge and 2 months after birth, the infant presented without any signs of neurologic sequelae, but further neurologic and developmental follow-up studies remain necessary. CONCLUSION: Acute subdural hematoma is a rare but severe intracranial complication of obstetric trauma. The early recognition of this complication and immediate surgical intervention could possibly prevent later neurologic sequelae. PMID- 12749660 TI - Near-fatal grape aspiration with complicating acute lung injury successfully treated with extracorporeal membrane oxygenation. AB - OBJECTIVE: In this report of a near-fatal case of grape aspiration successfully treated with extracorporeal membrane oxygenation (ECMO), we highlight the danger of feeding seedless grapes to young children and demonstrate that ECMO can provide cardiopulmonary support for cases of acquired large-airway disruption and can facilitate therapeutic intervention. DESIGN: Case report. SETTING: A tertiary pediatric intensive care unit and ECMO center. PATIENT: A healthy 14-month-old boy aspirated a seedless grape while playing at home and suffered a cardiopulmonary arrest of 15 mins in duration. He responded to advanced life support with return of cardiac output but developed intractable cardiopulmonary failure secondary to aspirated grape particles and postobstructive pulmonary edema. INTERVENTIONS: The patient was emergently transferred to the regional ECMO center and placed on venoarterial ECMO. Bronchoscopies were performed in the stable environment provided by ECMO, aspirated particles were removed from the large airways, and lung recovery was facilitated. MEASUREMENTS AND MAIN RESULTS: End-organ perfusion was restored via ECMO during a period of severe intractable cardiopulmonary failure. Pulmonary recovery occurred during a 6-day ECMO run and was facilitated by therapeutic bronchoscopy. The patient was reviewed 1 yr later and has made a full neurodevelopmental recovery, despite a 15-min out-of-hospital cardiac arrest. CONCLUSIONS: Aspiration of a seedless grape is a life-threatening event in a small child. This danger is not fully appreciated by parents in the UK. ECMO may be life saving in cases of acquired large-airway disruption resulting in severe cardiopulmonary failure, including foreign body aspiration, as long as end-organ perfusion is maintained. PMID- 12749662 TI - Successful management of an extreme example of neonatal hyperprostaglandin-E syndrome (Bartter's syndrome) with the new cyclooxygenase-2 inhibitor rofecoxib. AB - OBJECTIVE: To describe the successful treatment of an unusual case of severe neonatal Bartter's syndrome refractory to treatment with indomethacin. DESIGN: Case report, clinical. SETTING: Tertiary care intensive care unit. PATIENTS: A patient with neonatal hyperprostaglandin-E syndrome and excessive requirements of intravenous (via central venous catheter) water and salt supplementation, failure to thrive, vomiting, and massive growth retardation, despite adequate treatment with indomethacin. MAIN RESULT: Four weeks after induction of the new cyclooxygenase-2 inhibitor rofecoxib, the patient was well, on full enteral feeds, thriving, and had gained 600 g in weight. A lower supplementary potassium, magnesium, and sodium intake was required. Reinstitution of indomethacin therapy resulted in severe deterioration, despite high indomethacin doses; symptoms improved again after rofecoxib administration. No side effects have been seen thus far. CONCLUSION: This report shows that in selected patients with a severe form of neonatal Bartter's syndrome, the new cyclooxygenase-2 inhibitor rofecoxib may control the clinical symptoms of hyperprostaglandin-E syndrome after ineffective indomethacin therapy. PMID- 12749663 TI - Treatment of acute lymphoblastic leukemia-induced extreme hypercalcemia with pamidronate and calcitonin. AB - OBJECTIVE: To describe extreme hypercalcemia as the presenting feature of acute lymphoblastic leukemia in an 8-yr-old girl and the combined use of pamidronate and calcitonin for its treatment. DESIGN: Case report. SETTING: Pediatric intensive care unit. PATIENT: An 8-yr-old girl with 20.0 mg/dL serum calcium (reference range, 8.8-10.4 mg/dL) and 2.66 mmol/L ionized calcium (reference range, 1.13-1.32 mmol/L). INTERVENTION: Intravenous pamidronate and subcutaneous calcitonin. MEASUREMENTS AND MAIN RESULTS: Our patient presented with nausea, vomiting, lethargy, weight loss, fatigue, and weakness but, remarkably, did not exhibit electrocardiographic changes. Initial treatment with hydration at 8 mL x kg(-1) x hr(-1) and furosemide was ineffective. A single dose of 1 mg/kg intravenous pamidronate given over 24 hrs complemented by three doses of 5 units/kg subcutaneous calcitonin over 36 hrs lowered serum calcium to a normal range within 3 days. Side effects noted were hypocalcemia, hypomagnesemia, and hypophosphatemia. They were most pronounced 7-9 days after treatment, stabilized with supplementation, and returned to acceptable ranges by 1 month without need for ongoing electrolyte supplements. A renal computed tomographic scan did not show nephrocalcinosis. The patient remained free from recurrence of hypercalcemia 6 wks after initiating chemotherapy for acute lymphoblastic leukemia. CONCLUSIONS: Extreme hypercalcemia can be a presenting feature of acute lymphoblastic leukemia, but it may not result in life-threatening organ dysfunction. Combined treatment with pamidronate and calcitonin should be considered for treating hypercalcemia that does not respond to conventional therapy with hydration and furosemide. PMID- 12749664 TI - Prematurity-associated costs: more than just surfactant. PMID- 12749665 TI - Global paternalism in pediatric intensive care unit end-of-life decisions? PMID- 12749666 TI - Art of communication. PMID- 12749667 TI - Predicting outcome in childhood bacterial meningitis: are pneumococci stronger than glutamate? PMID- 12749668 TI - From energy expenditure to nutritional intervention? PMID- 12749669 TI - Prelude to pediatric multiple organ dysfunction syndrome: the golden hours concept revisited. PMID- 12749670 TI - Procalcitonin as a diagnostic and prognostic biomarker of sepsis in critically ill children. PMID- 12749671 TI - Introduction to apoptosis in ophthalmology. AB - Apoptosis represents a mode of cellular death genetically programmed to maintain homeostasis of tissues. In specific pathologic circumstances, the death program may be activated by various environmental factors such as exposure to toxic substances or bacteria or deprivation of nutrients. From this point of view, apoptosis is considered the final event in several pathologies. In ophthalmology, experimental evidence has confirmed that apoptosis is a type of cellular death involved in various pathologic processes including glaucoma, retinitis pigmentosa, ischemic retinopathy, corneal reparative processes, cataract, and retinoblastoma. The aim of this article is to review the most recent results published in this field and to describe some of the molecular mechanisms responsible for the activation of the apoptotic program in some important ocular disorders. The understanding of such mechanisms could outline new therapeutic strategies for the prevention of cellular death in ophthalmology. PMID- 12749672 TI - The mechanisms of apoptosis in biology and medicine: a new focus for ophthalmology. AB - Defects in apoptosis (programmed cell death) have recently emerged as being closely involved in the pathogenesis of most ocular diseases and, therefore, apoptosis is now a topic of exponential interest in ophthalmology. This review summarizes recent works on mechanisms of apoptosis, from its initiation and modulation to the switching-on of its execution machinery. Interactions of cell death with cell division programs to orchestrate ontogenesis, aging, and adult life and their alterations in human diseases are pointed out. Two main apoptotic signaling pathways are identified: a death receptor-dependent (extrinsic) pathway and a mitochondrion-dependent (intrinsic) pathway. Mitochondrion harbors both antiapoptotic (Bcl-2, Bcl-XL) and apoptotic factors (Smac/Diablo, Apaf-1, cytochrome c). Its permeability transition pore (mPTP) is the main trigger of cell suicide. The process of mPTP opening, in association with extrusion to cytoplasm of a variety of apoptotic factors, is shown. Cytochrome c is one of these apoptotic factors. When expelled to cytoplasm, this double-faced respiratory chain component assembles with two other modules, Apaf-1 and procaspase 9, to form a protein complex--the apoptosome--that starts apoptosis execution. Another respiratory chain component, the CoQ10, is believed to counteract mPTP opening. What makes apoptosis particularly exciting for medicine is that its dysfunctions play a central role in the pathogenesis of several human diseases. For instance, excesses of apoptosis lead to cell loss that accompanies neurodegenerative diseases, whereas genetically determined defects of apoptosis lead to the deregulated cell proliferation typical of cancer. A variety of ophthalmologic diseases, such as post-keratectomy haze, corneal lesions, cataract, glaucoma, senile maculopathies, and genetic ocular pathologies, that underlie apoptosis dysfunctions are treated in detail in the other reviews of this issue. PMID- 12749673 TI - Some current ideas on the pathogenesis and the role of neuroprotection in glaucomatous optic neuropathy. AB - The primary features of glaucomatous optic neuropathy are characteristic changes in the optic nerve head, a decrease in number of surviving ganglion cells and a reduction in vision. It is now generally accepted that a number of factors, including elevated intraocular pressure, could lead to the changes seen in the optic nerve head and to obtain a pharmacological means to treat the causes will vary from patient to patient. In contrast, a cascade of events have been proposed to explain how the changes in the optic nerve head may lead to the slow and differential death of ganglion cells in the disease. It is also proposed that drugs (neuroprotectants) influencing this cascade of events can attenuate ganglion cell death and lead to the treatment of all glaucoma patients. PMID- 12749674 TI - Neuroprotection as a treatment for glaucoma: pharmacological and immunological approaches. AB - Primary open-angle glaucoma is a chronic, progressive optic neuropathy associated with a gradual decline in visual function, which may lead to blindness. In most cases, the optic neuropathy is associated with increased intraocular pressure. It is now generally accepted, however, that normalization of pressure, although necessary, is often not-sufficient as a remedial measure. This is because of the existence of additional factors, some of which emerge as a consequence of the initial damage. This situation is reminiscent of the response to a traumatic axonal insult, in which some of the damage is immediate and is caused by the insult itself, and some is secondary and is caused by a deficiency of growth supportive factors as well as by toxic factors derived from the damaged tissue. Accordingly, the author has suggested that glaucoma may be viewed as a neurodegenerative disease and consequently amenable to any therapeutic intervention applicable to neurodegenerative diseases. There is evidence that neuroprotection can be achieved both pharmacologically and immunologically. Pharmacologic intervention neutralizes some of the effects of the nerve-derived toxic factors and possibly increases the ability of the remaining healthy neurons, at any given time, to cope with the stressful conditions. Immunologic intervention boosts the body's repair mechanisms for counteracting the toxicity of physiologic compounds acting as stress signals. PMID- 12749675 TI - Brain changes in glaucoma. AB - There is evidence that glaucomatous damage extends from retinal ganglion cells to vision centers in the brain. In the lateral geniculate nucleus (LGN), the major relay center between the eye and the visual cortex, neurons should undergo degenerative and/or neurochemical changes in magno-, parvo-, and koniocellular pathways conveying motion, red-green, and blue-yellow information, respectively. Furthermore, in both the LGN and visual cortex in glaucoma, changes in metabolic activity are observed. The study of brain changes in glaucoma may provide new insights into the pathobiology of glaucomatous damage and disease progression, and may stimulate new detection and therapeutic strategies to prevent blindness. PMID- 12749676 TI - Apoptosis in the mechanisms of neuronal plasticity in the developing visual system. AB - Visual experience during early postnatal life is essential for normal development of synaptic connections in the visual system. In fact, altered visual experiences such as monocular deprivation (MD) or abnormal visual stimulation (e.g. strabismus, anisometropia) during this period disrupt the physiologic organization of the visual pathway, leading to loss of visual responses in cortical neurons and reduction in visual acuity of the affected eye, so that it becomes amblyopic. The authors review the main functional and morphologic changes induced by altered visual experiences in the developing visual system and focus on the recent discovery that MD induces apoptotic cell death in the lateral geniculate nucleus of newborn rats. Particular attention is given to the authors' studies documenting that, during development, MD leads retinal terminals to release excessive glutamate in the lateral geniculate nucleus where it elevates nitric oxide and causes DNA fragmentation. The latter event is known to activate poly-(ADP-ribose) polymerase, which in turn may trigger apoptosis. Better understanding of the mechanisms underlying the morphologic changes induced by altered visual experiences during development may open new venues for studying novel neuroprotective strategies for amblyopia and, more generally, for the treatment of ophthalmic diseases associated with neuronal apoptosis. PMID- 12749677 TI - Pharmacological strategies to block rod photoreceptor apoptosis caused by calcium overload: a mechanistic target-site approach to neuroprotection. AB - PURPOSE: Photoreceptor apoptosis and resultant visual deficits occur in humans and animals with inherited, and disease-, injury- and chemical-induced retinal degeneration. Our aims were three-fold: 1) to determine the kinetics of rod apoptosis and Ca2+ overload in Pde6b9rd1) mice and developmentally lead-exposed rats, 2) to establish a pathophysiologically-relevant model of Ca2+ overload/rod selective apoptosis in isolated rat retina and 3) to examine different mechanistic based neuroprotective strategies that would abrogate or mollify rod Ca2+ overload/apoptosis. METHODS: Retinal morphometry and elemental calcium content ([Ca]) determined the kinetics of rod apoptosis and Ca2+ overload. A multiparametric analysis of apoptosis including rod [Ca], a live/dead assay, rod oxygen consumption, cytochrome c immunoblots and caspase assays was combined with pharmacological studies of an isolated rat retinal model of rod-selective Ca2+ overload/apoptosis. RESULTS: Ca2+ overload preceded rod apoptosis in mice and rats, although the extent and kinetics in each differed significantly. The isolated rat model of rod Ca2+ overload/apoptosis showed that blockade of Ca2+ entry through rod cGMP-activated channels with L-cis diltiazem was partially neuroprotective, whereas blockade of Ca2+ entry into rods through L-type Ca2+ channels with D-cis diltiazem or verapamil provided no protection. Inhibition of the mitochondrial Na+/Ca2+ exchanger with D-cis diltiazem provided no protection. CsA and NIM811, mitochondrial permeability transition pore (mPTP) inhibitors, blocked all Ca(2+)-induced apoptosis, whereas the caspase-3 inhibitor DEVD-fmk only blocked the downstream cytochrome c-induced apoptosis. CONCLUSIONS: The successful pharmacological neuroprotective strategies for rod Ca2+ overload/apoptosis targeted the rod cGMP-activated channels or mPTP, but not the rod L-type Ca2+ channels. PMID- 12749678 TI - Adequate nutrient intake can reduce cardiovascular disease risk in African Americans. AB - Cardiovascular disease kills nearly as many Americans each year as the next seven leading causes of death combined. The prevalence of cardiovascular disease and most of its associated risk factors is markedly higher and increasing more rapidly among African Americans than in any other racial or ethnic group. Improving these statistics may be simply a matter of improving diet quality. In recent years, a substantial and growing body of evidence has revealed that dietary patterns complete in all food groups, including nutrient-rich dairy products, are essential for preventing and reducing cardiovascular disease and the conditions that contribute to it. Several cardiovascular risk factors, including hypertension, insulin resistance syndrome, and obesity, have been shown to be positively influenced by dietary patterns that include adequate intake of dairy products. The benefits of nutrient-rich dietary patterns have been specifically tested in randomized, controlled trials emphasizing African American populations. These studies demonstrated proportionally greater benefits for African Americans without evidence of adverse effects such as symptoms of lactose intolerance. As currently promoted for the prevention of certain cancers and osteoporosis, regular consumption of diets that meet recommended nutrient intake levels might also be the most effective approach for reducing cardiovascular disease risk in African Americans. PMID- 12749679 TI - Nonstigmatizing ways to engage HIV-positive African-American teens in mental health and support services: a commentary. AB - This commentary illustrates a hospital/adolescent-clinic based model for providing support services and for increasing medical adherence among HIV positive inner city African-American adolescents. This commentary reviews the racial/ethnic disparities in HIV disease among adolescents and describes a successful program model for overcoming stigma. Traditional support groups were rejected by youth with HIV/AIDS. Seven elements common to successful programs were identified. Successful programs built on designs described in the research literature. Focus groups composed of HIV-positive adolescents identified what they wanted and needed. Stigmatizing labels were avoided in naming programs. Practical barriers to access, such as transportation and childcare, were eliminated. Programs were skills oriented, culturally sensitive, and life affirming, focusing on healthy living. HIV-positive inner city African-American youth can be successfully recruited and engaged in hospital based programs. Although these programs were qualitatively evaluated by youth as successful and attrition was low and attendance averaged 50%, rigorous quantitative research is needed to evaluate the effectiveness of such programs. We need quantitative research to successfully advocate for government funding. Stigma needs to be addressed openly in public health. Future research is needed to evaluate interventions to overcome the health consequences of stigma on utilization of available medical and mental health services. PMID- 12749680 TI - Can known risk factors explain racial differences in the occurrence of bacterial vaginosis? AB - BACKGROUND: Black women are more likely to have bacterial vaginosis (BV) than are non-Hispanic white women. We examined whether this disparity can be explained by racial differences in known BV risk factors. METHODS: Nine hundred black and 235 white women were enrolled from five US sites. At baseline, structured interviews were conducted and vaginal swabs self-collected for Gram-stain and culture. RESULTS: Black women were more likely than white women to have BV/intermediate vaginal flora. They also were more likely to be older, have lower educational attainment and family incomes, have a history of a sexually transmitted disease, and douche. After adjustment for demographic and lifestyle factors, blacks remained at elevated risk for BV/intermediate flora (OR 2.2, 95% CI 1.5-3.1). Blacks also were more likely to have specific BV-related vaginal microflora, as well as gonococcal or chlamydial cervicitis (OR 2.2, 95% CI 1.2-3.8) after adjustment for known BV risk factors. CONCLUSION: Risk factor differences did not explain the observed racial disparity in the occurrence of BV, BV-related microflora, or gonococcal or chlamydial cervicitis. These findings highlight our limited understanding of the factors accounting for the occurrence of bacterial vaginosis and cervicitis among black and white women. PMID- 12749682 TI - Primary renal non-Hodgkins lymphoma presenting with acute renal failure. AB - Primary renal lymphoma (PRL) has been reported in medical literature. Its occurrence is rare and controversial, the kidney being an extranodal organ. We report a case of primary renal lymphoma presenting with acute-on-chronic renal failure and unilateral involvement of the left kidney without obstruction and with minimal peripheral organ involvement. Definitive diagnosis was made from histologic examination of the mass postoperatively. Renal function became stabilized after the removal of the tumor. PMID- 12749683 TI - Enslaved Africans and doctors in South Carolina. AB - This interpretation of the relationship between enslavement and American medicine in 19th century South Carolina reveals the intimacy that existed between Africans enslaved in that state and the doctors who practiced and taught there. Enslaved Africans were resourceful and reliable medical figures in the slave community. Their knowledge of medical botany permeated the slave quarters and plantation hospitals and was appropriated into southern medical knowledge. The trajectories of the careers of three South Carolina physicians are tied to their practice around and on the enslaved. The beginnings of gynecological surgery are linked to 1840s experimentation on enslaved African women performed by one of them. PMID- 12749681 TI - Congenital cytomegalovirus infection. AB - Cytomegalovirus (CMV) is the most common congenital infection in humans. Congenital CMV infection can follow either a primary or recurrent maternal infection, but the likelihood of fetal infection and the risk of associated damage is higher after a primary infection. Approximately 90% of congenitally infected infants are asymptomatic at birth. Jaundice, petechiae, and hepatosplenomegaly are the most frequently noted clinical triad in symptomatic infants. More frequent and more severe sequelae occur in symptomatic infants, notably psychomotor hearing loss and retardation. Congenital CMV infection can be diagnosed by isolation of the virus from the urine or saliva within the first three weeks of life. Rapid diagnosis can be accomplished by detection of CMV DNA by DNA amplification or hybridization techniques. PMID- 12749685 TI - A need for medical awareness in black society. PMID- 12749684 TI - Harlem health care, a look back. An interview with Edward A. Nichols, MD/pediatrician. Interview by George A. Dawson. AB - Our goal in this series of interviews is to provide a historical record, of sorts, and to highlight persons of African American ancestry who are health care professionals in Harlem, New York. Today, Harlem is undergoing a second cultural renaissance, and in this milieu, doctors are active contributors as providers of health care and, therefore, must be reckoned with as the proverbial backbone to any community change, be it positive or negative. In this instance, we judge the changes occurring in Harlem, for the most part, to be positive. Our inaugural interview is with Dr. Nichols, a longtime member of the National Medical Association and a pediatrician who has practiced in Harlem for well over 25 years. PMID- 12749686 TI - The Enlightenment and the many faces of Fundamentalism. PMID- 12749687 TI - Mammalian phosphatidylinositol 4-kinases. AB - Three phosphatidylinositol 4-kinase isoforms, PI4K 230, 92 and 55 have been cloned and sequenced allowing a much wider characterization than the previously employed enzymological typing into type II and III enzymes. PI4K 230 and 92 contain a highly conserved catalytic core, PI4K55 one with a much lower degree of similarity. Candidate kinase motifs, deduced from the protein kinase super family, are absolutely conserved in all isoforms. Kinase activities are described based on their sensitivity and reactivity towards wortmannin, phenylarsine oxide (PAO) and 5'-p-fluorosulfonylbenzoyladenosine (FSBA). Localization of all isoforms in the cell is reported. All enzymes contain nuclear localization and export sequence motifs (NLS and NES) leading to the expectation that they can be transferred to the nucleus. PI4K230 has been found in the nucleolus, PI4K92 in the nucleus, additionally further broadening the function of these enzymes. In the cytoplasm of neuronal cells, PI4K230 is distributed evenly on membranes that are ultra structurally cisterns of the rough endoplasmatic reticulum, outer membranes of mitochondria, multivesicular bodies, and are in close vicinity of synaptic contacts. PI4K92 is functionally characterized as a key enzyme regulating Golgi disintegration/reorganization during mitosis probably via phosphorylation by cyclin-dependent kinases on well-defined sites. PI4K55 is involved in the production of second messengers, diacylglycerol (DAG) and inositol 1,4,5-trisphosphate (InsP3) at the plasma membrane, moreover, in the endocytotic pathway in the cytoplasm. PMID- 12749688 TI - Anti-retroviral protease inhibitors--'a two edged sword?'. AB - The use of anti-retroviral protease inhibitors in combination with nucleoside analog or non nucleoside reverse transcriptase inhibitors (HAART) has led to dramatic decreases in the mortality seen with HIV infected patients. In concert with these treatment regimens, especially with the inclusion of the anti retroviral protease inhibitors (PI), a complex series of metabolic complications occurred. These included alterations of fat and carbohydrate metabolism. In some patients, one could observe either lipoatrophy (fat wasting) as well as lipohypertrophy (fat deposition) or both. The problem is that the lack of a case definition of the altered fat metabolism confuses diagnoses. In vitro, interference with fat cell differentiation has been demonstrated by PI. Further, in vitro studies demonstrate that indinavir, a PI currently used in HIV treatment, can interact with the insulin responsive glucose transporter (GLUT4). The activity of the GLUT4 is inhibited by indinavir and eventual insulin resistance has been shown (i.e. in vivo and in vitro). Also, controversy exists regarding insulin signaling in fat cells. Finally, the relationships between hyperlipidemia and/or lipolysis and altered carbohydrate metabolism (i.e. mild glucose intolerance, insulin resistance) suggest an association with cardiovascular risk in protease treated patients (Metabolic Syndrome X). In short, while multiple problems exist, no one mechanism can account for the changes observed. PMID- 12749689 TI - Plant peroxisomes, reactive oxygen metabolism and nitric oxide. AB - In plant cells, as in most eukaryotic organisms, peroxisomes are probably the major sites of intracellular H2O2 production, as a result of their essentially oxidative type of metabolism. Like mitochondria and chloroplasts, peroxisomes also produce superoxide radicals (O2*-) and there are, at least, two sites of superoxide generation: one in the organelle matrix, the generating system being xanthine oxidase, and another site in the peroxisomal membranes dependent on NAD(P)H. In peroxisomal membranes, three integral polypeptides (PMPs) with molecular masses of 18, 29, and 32 kDa have been shown to generate O2*- radicals. Besides catalase, several antioxidative systems have been demonstrated in plant peroxisomes, including different superoxide dismutases, the four enzymes of the ascorbate-glutathione cycle plus ascorbate and glutathione, and three NADP dependent dehydrogenases. A CuZn-SOD and two Mn-SODs have been purified and characterized from different types of plant peroxisomes. The presence of the enzyme nitric oxide synthase (NOS) and its reaction product, nitric oxide (NO*), has been recently demonstrated in plant peroxisomes. Different experimental evidence has suggested that peroxisomes have a ROS-mediated cellular function in leaf senescence and in stress situations induced by xenobiotics and heavy metals. Peroxisomes could also have a role in plant cells as a source of signal molecules like NO*, O2*- radicals, H2O2, and possibly S-nitrosoglutathione (GSNO). It seems reasonable to think that a signal molecule-producing function similar to that postulated for plant peroxisomes could also be performed by human, animal and yeast peroxisomes, where research on oxy radicals, antioxidants and nitric oxide is less advanced than in plant peroxisomes. PMID- 12749690 TI - Yeast models of human mitochondrial diseases. AB - The yeast Saccharomyces cerevisiae is an excellent model for gaining insights into the molecular basis of human mitochondrial disorders, particularly those resulting from impaired mitochondrial metabolism. Yeast is a very well characterized system and most of our current knowledge about mitochondrial biogenesis in humans derives from yeast genetics and biochemistry. Systematic yeast genome-wide approaches have allowed for the identification of human disease genes. In addition, the functional characterization of a large number of yeast gene products resident in mitochondria has been instrumental for the later identification and characterization of their human orthologs. Here I will review the molecular and biochemical characterization of several mitochondrial diseases that have been ascribed to mutations in genes that were first found in yeast to be necessary for the assembly of the mitochondrial respiratory chain. The usefulness of yeast as a model system for human mitochondrial disorders is evaluated. PMID- 12749691 TI - Impedance of Helicobacter pylori lipopolysaccharide interference with gastric mucin synthesis by peroxisome proliferator-activated receptor gamma activation involves phosphatidylinositol 3-kinase/ERK pathway. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) plays a critical role in the regulation of the expression of genes associated with inflammation. In this study, we report that PPARgamma activation leading to the impedance of H. pylori lipopolysaccharide (LPS) inhibitory effect on gastric mucin synthesis occurs with the involvement of phosphatidylinositol 3-kinase (PI3K) and extracellular signal-regulated kinase (ERK) pathways. Using gastric mucosal cells in culture, we show that activation of PPARgamma with a specific synthetic agonist, ciglitazone, prevents in a dose-dependent fashion (up to 90.2%) the LPS induced reduction in mucin synthesis, and the effect is reflected in a marked decrease in the LPS-induced apoptosis (72.4%), NO generation (80.1%), and the expression of NOS-2 activity (90%). The impedance by ciglitazone of the LPS induced reduction in mucin synthesis was blocked by wortmannin, a specific inhibitor of P13K and PD98059, an inhibitor of ERK. Both inhibitors, moreover, caused further enhancement in the LPS-induced NO generation and countered the inhibitory effect of ciglitazone on the LPS-induced upregulation in NOS-2. Our findings point to PI3K and ERK as mediators of PPARgamma agonist effect leading to the impedance of H. pylori LPS inhibition on gastric mucin synthesis. PMID- 12749692 TI - Isoelectric point mobility shift assay for rapid screening of charged and uncharged ligands bound to proteins. AB - Three human proteins (hTAP1, hTAP2 and hTAP3) that are related to the yeast phosphatidylinositol/phosphatidylcholine transfer protein SEC14p were recently cloned in our laboratory. These proteins contain a relatively large hydrophobic pocket, the so called CRAL-TRIO domain, which is present also in other human proteins, such as CRALBP, alpha-TTP and MEG2. The CRAL-TRIO domains in these proteins bind ligands such as retinaldehyde, tocopherols and polyphosphoinositides, respectively. To screen for potential hTAPs ligands, we developed a semi-quantitative isoelectric point mobility shift assay (IPMS-assay) that allows assessing the binding of potential hydrophobic ligands to proteins. Purified proteins occupied with a charged ligand migrate differently on isoelectric focusing gels when compared with free protein. Competition of bound charged ligands with uncharged ones reverses the mobility shift, so that the relative affinities of the two ligands to the protein can be estimated. PMID- 12749693 TI - The maximum size of glycogen molecules. PMID- 12749694 TI - How I became a biochemical pharmacologist. PMID- 12749696 TI - Regulation of membrane transport by rab GTPases. AB - Membrane flow through the cell is a highly dynamic process in which intracellular compartments communicate via tubulo-vesicular structures shuttling cargo molecules to their destinations. Transport carriers are formed at a donor compartment and navigate through the cytoplasm to the target organelle, on which they subsequently dock and fuse. Many of these events are regulated by the cooperative action of monomeric rab GTPases and their effector proteins. Research in recent years resulted in the identification of many rab effectors, providing first glimpses how the GTPase switch of individual rab proteins is utilized in discrete transport steps. PMID- 12749695 TI - Aspartic peptidase inhibitors: implications in drug development. AB - The last decade has witnessed an effervescence of research interest in the development of potent inhibitors of various aspartic peptidases. As an enzyme family, aspartic peptidases are relatively a small group that has received enormous interest because of their significant roles in human diseases like involvement of renin in hypertension, cathepsin D in metastasis of breast cancer, beta-Secretase in Alzheimer's Disease, plasmepsins in malaria, HIV-1 peptidase in acquired immune deficiency syndrome, and secreted aspartic peptidases in candidal infections. There have been developments on clinically active inhibitors of HIV-1 peptidase, which have been licensed for the treatment of AIDS. The inhibitors of plasmepsins and renin are considered a viable therapeutic strategy for the treatment of malaria and hypertension. Relatively few inhibitors of cathepsin D have been reported, partly because of its uncertain role as a viable target for therapeutic intervention. The beta-secretase inhibitors OM99-2 and OM003 were designed based on the substrate specificity information. The present article is a comprehensive state-of-the-art review describing the aspartic peptidase inhibitors illustrating the recent developments in the area. In addition, the homologies between the reported inhibitor sequences have been analyzed. The understanding of the structure-function relationships of aspartic peptidases and inhibitors will have a direct impact on the design of new inhibitor drugs. PMID- 12749697 TI - Analysis of structure and function of putative surface-exposed proteins encoded in the Streptococcus pneumoniae genome: a bioinformatics-based approach to vaccine and drug design. AB - Streptococcus pneumoniae is the most common cause of fatal community-acquired pneumonia, middle ear infection, and meningitis. The prevention and treatment of this infection have become a top priority for the medical-scientific community. The present polysaccharide-based vaccine used to immunize susceptible hosts is only approximately 60% effective and is ineffective in children younger than 2 years of age. The new conjugate vaccine, based on the engineered diphtheria toxin coupled to polysaccharide antigens. is approved only for use in children under 2 years of age to treat invasive disease. While penicillin is the drug of choice to treat infections secondary to S. pneumoniae, increasing numbers of bacterial strains are resistant to penicillin as well as to broad spectrum antibiotics such as vancomycin. Thus, there is a need to identify new strategies to prevent and treat diseases caused by to S. pneumoniae. In this article, we summarize the utilization of the recently available S. pneumoniae genomic information in order to identify and characterize novel proteins likely located on the surface of this Gram-positive pathogenic bacterium. Because only a limited number of surface proteins of S. pneumoniae have been characterized to date, this information provides new insights into the pathogenesis of this organism as well as highlights possible avenues for its treatment and/or prevention in the future. The review is divided into two sections. First, we brietly summarize current information about known surface-exposed proteins of S. pneumoniae. This is followed by the illustration of procedures for the identification of new putative surface-exposed proteins. These have signal peptides required for their extra cytoplasmic transport and/or additional signature sequences. Some of these will be S. pneumoniae virulence factors. The signature sequences we have chosen are those leading to protein binding to choline present on the bacterial surface, attachment to peptidoglycan of the cell wall, or anchoring to lipids of the cytoplasmic membrane. All these signatures are indicative of binding of proteins to the surface of this organism. Secondly, we illustrate the application of bioinformatics and modeling tools to these selected proteins in order to provide information about their likely functions and preliminary three-dimensional structure models. The focal point of the analysis of these proteins, their sequences, and structures is the evaluation of their antigenic properties and possible roles in pathogenicity. The information obtained from the genome analysis will be instrumental in the development of a more effective prophylactic and/or therapeutic agents to prevent and to treat infections due to S. pneumoniae. PMID- 12749698 TI - A critical assessment of boron neutron capture therapy: an overview. AB - Boron neutron capture therapy (BNCT) is based on the nuclear reaction that occurs when boron-10 is irradiated with neutrons of the appropriate energy to produce high-energy alpha particles and recoiling lithium-7 nuclei. BNCT has been used clinically to treat patients with high-grade gliomas, and a much smaller number with primary and metastatic melanoma. The purpose of this special issue of the Journal of Neuro-Oncology is to provide a critical and realistic assessment of various aspects of basic and clinical BNCT research in order to better understand its present status and future potential. Topics that are covered include neutron sources, tumor-targeted boron delivery agents, brain tumor models to assess therapeutic efficacy, computational dosimetry and treatment planning, results of clinical trails in the United States, Japan and Europe, pharmacokinetic studies of sodium borocaptate and boronophenylalanine (BPA), positron emission tomography imaging of BPA for treatment planning, and finally an overview of the challenges and problems that must be faced if BNCT is to become a useful treatment modality for brain tumors. Clinical studies have demonstrated the safety of BNCT. The next challenge is an unequivocal demonstration of therapeutic efficacy in one or more of the clinical trails that either are in progress or are planned over the next few years. PMID- 12749699 TI - Fission reactor neutron sources for neutron capture therapy--a critical review. AB - The status of fission reactor-based neutron beams for neutron capture therapy (NCT) is reviewed critically. Epithermal neutron beams, which are favored for treatment of deep-seated tumors, have been constructed or are under construction at a number of reactors worldwide. Some of the most recently constructed epithermal neutron beams approach the theoretical optimum for beam purity. Of these higher quality beams, at least one is suitable for use in high through-put routine therapy. It is concluded that reactor-based epithermal neutron beams with near optimum characteristics are currently available and more can be constructed at existing reactors. Suitable reactors include relatively low power reactors using the core directly as a source of neutrons or a fission converter if core neutrons are difficult to access. Thermal neutron beams for NCT studies with small animals or for shallow tumor treatments, with near optimum properties have been available at reactors for many years. Additional high quality thermal beams can also be constructed at existing reactors or at new, small reactors. Furthermore, it should be possible to design and construct new low power reactors specifically for NCT, which meet all requirements for routine therapy and which are based on proven and highly safe reactor technology. PMID- 12749700 TI - Accelerator-based epithermal neutron sources for boron neutron capture therapy of brain tumors. AB - This paper reviews the development of low-energy light ion accelerator-based neutron sources (ABNSs) for the treatment of brain tumors through an intact scalp and skull using boron neutron capture therapy (BNCT). A major advantage of an ABNS for BNCT over reactor-based neutron sources is the potential for siting within a hospital. Consequently, light-ion accelerators that are injectors to larger machines in high-energy physics facilities are not considered. An ABNS for BNCT is composed of: (1) the accelerator hardware for producing a high current charged particle beam, (2) an appropriate neutron-producing target and target heat removal system (HRS), and (3) a moderator/reflector assembly to render the flux energy spectrum of neutrons produced in the target suitable for patient irradiation. As a consequence of the efforts of researchers throughout the world, progress has been made on the design, manufacture, and testing of these three major components. Although an ABNS facility has not yet been built that has optimally assembled these three components, the feasibility of clinically useful ABNSs has been clearly established. Both electrostatic and radio frequency linear accelerators of reasonable cost (approximately 1.5 M dollars) appear to be capable of producing charged particle beams, with combinations of accelerated particle energy (a few MeV) and beam currents (approximately 10 mA) that are suitable for a hospital-based ABNS for BNCT. The specific accelerator performance requirements depend upon the charged particle reaction by which neutrons are produced in the target and the clinical requirements for neutron field quality and intensity. The accelerator performance requirements are more demanding for beryllium than for lithium as a target. However, beryllium targets are more easily cooled. The accelerator performance requirements are also more demanding for greater neutron field quality and intensity. Target HRSs that are based on submerged-jet impingement and the use of microchannels have emerged as viable target cooling options. Neutron fields for reactor-based neutron sources provide an obvious basis of comparison for ABNS field quality. This paper compares Monte Carlo calculations of neutron field quality for an ABNS and an idealized standard reactor neutron field (ISRNF). The comparison shows that with lithium as a target, an ABNS can create a neutron field with a field quality that is significantly better (by a factor of approximately 1.2, as judged by the relative biological effectiveness (RBE)-dose that can be delivered to a tumor at a depth of 6cm) than that for the ISRNF. Also, for a beam current of 10 mA, the treatment time is calculated to be reasonable (approximately 30 min) for the boron concentrations that have been assumed. PMID- 12749701 TI - A critical assessment of boron target compounds for boron neutron capture therapy. AB - Boron neutron capture therapy (BNCT) has undergone dramatic developments since its inception by Locher in 1936 and the development of nuclear energy during World War II. The ensuing Cold War spawned the entirely new field of polyhedral borane chemistry, rapid advances in nuclear reactor technology and a corresponding increase in the number to reactors potentially available for BNCT. This effort has been largely oriented toward the eradication of glioblastoma multiforme (GBM) and melanoma with reduced interest in other types of malignancies. The design and synthesis of boron-10 target compounds needed for BNCT was not channeled to those types of compounds specifically required for GBM or melanoma. Consequently, a number of potentially useful boron agents are known which have not been biologically evaluated beyond a cursory examination and only three boron-10 enriched target species are approved for human use following their Investigational New Drug classification by the US Food and Drug Administration; BSH, BPA and GB-10. All ongoing clinical trials with GBM and melanoma are necessarily conducted with one of these three species and most often with BPA. The further development of BNCT is presently stalled by the absence of strong support for advanced compound evaluation and compound discovery driven by recent advances in biology and chemistry. A rigorous demonstration of BNCT efficacy surpassing that of currently available protocols has yet to be achieved. This article discusses the past history of compound development, contemporary problems such as compound classification and those problems which impede future advances. The latter include means for biological evaluation of new (and existing) boron target candidates at all stages of their development and the large-scale synthesis of boron target species for clinical trials and beyond. The future of BNCT is bright if latitude is given to the choice of clinical disease to be treated and if a recognized study demonstrating improved efficacy is completed. Eventually, BNCT in some form will be commercialized. PMID- 12749704 TI - Computational dosimetry and treatment planning considerations for neutron capture therapy. AB - Specialized treatment planning software systems are generally required for neutron capture therapy (NCT) research and clinical applications. The standard simplifying approximations that work well for treatment planning computations in the case of many other modalities are usually not appropriate for application to neutron transport. One generally must obtain an explicit three-dimensional numerical solution of the governing transport equation, with energy-dependent neutron scattering completely taken into account. Treatment planning systems that have been successfully introduced for NCT applications over the past 15 years rely on the Monte Carlo stochastic simulation method for the necessary computations, primarily because of the geometric complexity of human anatomy. However, historically, there has also been interest in the application of deterministic methods, and there have been some practical developments in this area. Most recently, interest has turned toward the creation of treatment planning software that is not limited to any specific therapy modality, with NCT as only one of several applications. A key issue with NCT treatment planning has to do with boron quantification, and whether improved information concerning the spatial biodistribution of boron can be effectively used to improve the treatment planning process. Validation and benchmarking of computations for NCT are also of current developmental interest. Various institutions have their own procedures, but standard validation models are not yet in wide use. PMID- 12749703 TI - Rat brain tumor models to assess the efficacy of boron neutron capture therapy: a critical evaluation. AB - Development of any therapeutic modality can be facilitated by the use of the appropriate animal models to assess its efficacy. This report primarily will focus on our studies using the F98 and 9L rat glioma models to evaluate the effectiveness of boron neutron capture therapy (BNCT) of brain tumors. Following intracerebral implantation the biological behavior of each tumor resembles that of human high grade gliomas in a number of ways. In both models, glioma cells were implanted intracerebrally into syngeneic Fischer rats and approximately 10 14 days later BNCT was initiated at the Brookhaven National Laboratory Medical Research Reactor. Two low molecular weight (M(r) < 210Da) 10B-containing drugs, boronophenylalanine (BPA) and/or sodium borocaptate (BSH) were used as capture agents, either alone or in combination with each other. The 9L gliosarcoma, which has been difficult to cure by means of either chemo- or radiotherapy alone, was readily curable by BNCT. The best survival data were obtained using BPA at a dose of 1200 mg/kg (64.8mg 10B), administered intraperitoneally (i.p.), with a 100% survival rate at 8 months. In contrast, the F98 glioma has been refractory to all therapeutic modalities. Tumor bearing animals, which had received 500 mg/kg (27 mg 10B) of BPA, or an equivalent amount of BSH i.v., had mean survival time (MST) of 37 and 33 days, respectively, compared to 29 days for irradiated controls. The best survival data with the F98 glioma model were obtained using BPA + BSH in combination, administered intra-arterially via the internal carotid artery (i.c.) with hyperosmotic mannitol induced blood-brain barrier disruption (BBB-D). The MST was 140 days with a cure rate of 25%, compared to a MST of 73 days with a 5% cure rate without BBB-D, and 41 days following i.v. administration of both drugs. A modest but significant increase in MST also was observed in rats that received intracarotid (i.c.) BPA in combination with Cereport (RMP-7), which produced a pharmacologically mediated opening of the BBB. Studies also have been carried out with the F98 glioma to determine whether an X-ray boost could enhance the efficacy of BNCT, and it was shown that there was a significant therapeutic gain. Finally, molecular targeting of the epidermal growth factor receptor (EGFR) has been investigated using F98 glioma cells, which had been transfected with the gene encoding EGFR and, intratumoral injection of boronated EGF as the delivery agent, followed by BNCT. These studies demonstrated that there was specific targeting of EGFR and provided proof of principle for the use of high molecular weight, receptor targeting-boron delivery agents. Finally, a xenograft model for melanoma metastatic to the brain has been developed using a human melanoma (MRA27), stereotactically implanted into the brains of nude rats, and these studies demonstrated that BNCT either cured or significantly prolonged the survival of tumor-bearing rats. It remains to be determined, which, if any, of these experimental approaches will be translated into clinical studies. Be that as it may, rat brain tumor models already have made a significant contribution to the design of clinical BNCT protocols, and should continue to do so in the future. PMID- 12749702 TI - Ligand liposomes and boron neutron capture therapy. AB - Boron neutron capture therapy (BNCT) has been used both experimentally and clinically for the treatment of gliomas and melanomas, with varying results. However, the therapeutic effects on micro-invasive tumor cells are not clear. The two drugs that have been used clinically, p-boronophenylalanine, (BPA), and the sulfhydryl borane, (BSH), seem to be taken up preferentially in solid tumor areas but it is uncertain whether enough boron is taken up by micro-invasive tumor cells. To increase the selective uptake of boron by such cells, would be to exploit tumor transformation related cellular changes such as over-expression of growth factor receptors. However, the number of receptors varies from small to large and the uptake of large amounts of boron for each receptor interaction is necessary in order to deliver sufficient amounts of boron. Therefore, each targeting moiety must deliver large number of boron atoms. One possible way to meet these requirements would be to use receptor-targeting ligand liposomes, containing large number of boron atoms. This will be the subject of this review and studies of boron containing liposomes, with or without ligand, will be discussed. Two recent examples from the literature are ligand liposomes targeting either folate or epidermal growth factor (EGF) receptors on tumor cells. Other potential receptors on gliomas include PDGFR and EGFRvIII. Besides the appropriate choice of target receptor, it is also important to consider delivery of the ligand liposomes, their pharmacodynamics and pharmacokinetics and cellular processing, subjects that also will be discussed in this review. PMID- 12749705 TI - Clinical review of the Japanese experience with boron neutron capture therapy and a proposed strategy using epithermal neutron beams. AB - Our concept of boron neutron capture therapy (BNCT) is selective destruction of tumor cells using the heavy-charged particles yielded through 10B(n, alpha)7 Li reactions. To design a new protocol that employs epithermal neutron beams in the treatment of glioma patients, we examined the relationship between the radiation dose, histological tumor grade, and clinical outcome. Since 1968, 183 patients with different kinds of brain tumors were treated by BNCT; for this retrospective study, we selected 105 patients with glial tumors who were treated in Japan between 1978 and 1997. In the analysis of side effects due to radiation, we included all the 159 patients treated between 1977 and 2001. With respect to the radiation dose (i.e. physical dose of boron n-alpha reaction), the new protocol prescribes a minimum tumor volume dose of 15 Gy or, alternatively, a minimum target volume dose of 18 Gy. The maximum vascular dose should not exceed 15 Gy (physical dose of boron n-alpha reaction) and the total amount of gamma rays should remain below 10 Gy, including core gamma rays from the reactor and capture gamma in brain tissue. The outcomes for 10 patients who were treated by the new protocol using a new mode composed of thermal and epithermal neutrons are reported. PMID- 12749706 TI - Assessment of the results from the phase I/II boron neutron capture therapy trials at the Brookhaven National Laboratory from a clinician's point of view. AB - Boron neutron capture therapy (BNCT) represents a promising modality for a relatively selective radiation dose delivery to the tumor tissue. The key to effective BNCT of tumors such as glioblastoma multiforme (GBM) is the homogeneous preferential accumulation of 10B in the tumor, including the infiltrating GBM cells, as compared to that in the vital structures of the normal brain. Provided that sufficiently high tumor 10B concentration (approximately 10(9) boron-10 atoms/cell) and an adequate thermal neutron fluence (approximately 10(9) neutrons/cm2) are achieved, it is the ratio of the 10B concentration in tumor cells to that in the normal brain cells and the blood that will largely determine the therapeutic gain of BNCT. PMID- 12749707 TI - A critical examination of the results from the Harvard-MIT NCT program phase I clinical trial of neutron capture therapy for intracranial disease. AB - A phase I trial was designed to evaluate normal tissue tolerance to neutron capture therapy (NCT); tumor response was also followed as a secondary endpoint. Between July 1996 and May 1999, 24 subjects were entered into a phase I trial evaluating cranial NCT in subjects with primary or metastatic brain tumors. Two subjects were excluded due to a decline in their performance status and 22 subjects were irradiated at the MIT Nuclear Reactor Laboratory. The median age was 56 years (range 24-78). All subjects had a pathologically confirmed diagnosis of either glioblastoma (20) or melanoma (2) and a Karnofsky of 70 or higher. Neutron irradiation was delivered with a 15 cm diameter epithermal beam. Treatment plans varied from 1 to 3 fields depending upon the size and location of the tumor. The 10B carrier, L-p-boronophenylalanine-fructose (BPA-f), was infused through a central venous catheter at doses of 250 mg kg(-1) over 1 h (10 subjects), 300 mg kg(-1) over 1.5 h (two subjects), or 350 mg kg(-1) over 1.5-2 h (10 subjects). The pharmacokinetic profile of 10B in blood was very reproducible and permitted a predictive model to be developed. Cranial NCT can be delivered at doses high enough to exhibit a clinical response with an acceptable level of toxicity. Acute toxicity was primarily associated with increased intracranial pressure; late pulmonary effects were seen in two subjects. Factors such as average brain dose, tumor volume, and skin, mucosa, and lung dose may have a greater impact on tolerance than peak dose alone. Two subjects exhibited a complete radiographic response and 13 of 17 evaluable subjects had a measurable reduction in enhanced tumor volume following NCT. PMID- 12749709 TI - Boron neutron capture therapy for glioblastoma multiforme: clinical studies in Sweden. AB - A boron neutron capture therapy (BNCT) facility has been constructed at Studsvik, Sweden. It includes two filter/moderator configurations. One of the resulting neutron beams has been optimized for clinical irradiations with a filter/moderator system that allows easy variation of the neutron spectrum from the thermal to the epithermal energy range. The other beam has been designed to produce a large uniform field of thermal neutrons for radiobiological research. Scientific operations of the Studsvik BNCT project are overseen by the Scientific Advisory Board comprised of representatives of major universities in Sweden. Furthermore, special task groups for clinical and preclinical studies have been formed to facilitate collaboration with academia. The clinical Phase II trials for glioblastoma are sponsored by the Swedish National Neuro-Oncology Group and, presently, involve a protocol for BNCT treatment of glioblastoma patients who have not received any therapy other than surgery. In this protocol, p boronophenylalanine (BPA), administered as a 6-h intravenous infusion, is used as the boron delivery agent. As of January 2002, 17 patients were treated. The 6-h infusion of 900 mg BPA/kg body weight was shown to be safe and resulted in the average blood-boron concentration of 24 microg/g (range: 15-32 microg/g) at the time of irradiation (approximately 2-3 h post-infusion). Peak and average weighted radiation doses to the brain were in the ranges of 8.0-15.5 Gy(W) and 3.3-6.1 Gy(W), respectively. So far, no severe BNCT-related acute toxicities have been observed. Due to the short follow-up time, it is too early to evaluate the efficacy of these studies. PMID- 12749708 TI - Boron neutron capture therapy of brain tumors: clinical trials at the finnish facility using boronophenylalanine. AB - Two clinical trials are currently running at the Finnish dedicated boron neutron capture therapy (BNCT) facility. Between May 1999 and December 2001, 18 patients with supratentorial glioblastoma were treated with boronophenylalanine (BPA) based BNCT within a context of a prospective clinical trial (protocol P-01). All patients underwent prior surgery, but none had received conventional radiotherapy or cancer chemotherapy before BNCT. BPA-fructose was given as 2-h infusion at BPA dosages ranging from 290 to 400 mg/kg prior to neutron beam irradiation, which was given as a single fraction from two fields. The average planning target volume dose ranged from 30 to 61 Gy (W), and the average normal brain dose from 3 to 6 Gy (W). The treatment was generally well tolerated, and none of the patients have died during the first months following BNCT. The estimated 1-year overall survival is 61%. In another trial (protocol P-03), three patients with recurring or progressing glioblastoma following surgery and conventional cranial radiotherapy to 50-60 Gy, were treated with BPA-based BNCT using the BPA dosage of 290 mg/kg. The average planning target dose in these patients was 25-29 Gy (W), and the average whole brain dose 2-3 Gy (W). All three patients tolerated brain reirradiation with BNCT, and none died during the first three months following BNCT. We conclude that BPA-based BNCT has been relatively well tolerated both in previously irradiated and unirradiated glioblastoma patients. Efficacy comparisons with conventional photon radiation are difficult due to patient selection and confounding factors such as other treatments given, but the results support continuation of clinical research on BPA-based BNCT. PMID- 12749710 TI - Tissue uptake of BSH in patients with glioblastoma in the EORTC 11961 phase I BNCT trial. AB - PURPOSE: The uptake of the boron compound Na2B12H10-SH (BSH) in tumor and normal tissues was investigated in the frame of the EORTC phase I trial 'Postoperative treatment of glioblastoma with BNCT at the Petten Irradiation Facility' (protocol 11961). METHODS AND MATERIALS: The boron concentration in blood, tumor, normal brain, dura, muscle, skin and bone was detected using inductively coupled plasma atomic emission spectroscopy in 13 evaluable patients. In a first group of 10 patients 100 mg BSH/kg bodyweight (BW) were administered; a second group of 3 patients received 22.9 mg BSH/kg BW. The toxicity due to BSH was evaluated. RESULTS: The average boron concentration in the tumor was 19.9 +/- 9.1 ppm (1 standard deviation (SD)) in the high dose group and 9.8 +/- 3.3 ppm in the low dose group, the tumor/blood ratios were 0.6 +/- 0.2 and 0.9 +/- 0.2, respectively. The highest boron uptake has been detected in the dura, very low uptake was found in the bone, the cerebro-spinal fluid and especially in the brain (brain/blood ratio 0.2 +/- 0.02 and 0.4 +/- 0.2). No toxicity was detected except flush-like symptoms in 2 cases during a BSH infusion at a much higher speed than prescribed. CONCLUSION: BSH proved to be safe for clinical application at a dose of 100 mg BSH/kg infused and at a dose rate of 1 mg/kg/min. The study underlines the importance of a further investigation of BSH uptake in order to obtain enough data for significant statistical analysis. The boron concentration in blood seems to be a quite reliable parameter to predict the boron concentration in other tissues. PMID- 12749712 TI - Pharamacokinetic modeling for boronophenylalanine-fructose mediated neutron capture therapy: 10B concentration predictions and dosimetric consequences. AB - A two-compartment open model has been developed for predicting 10B concentrations in blood following intravenous infusion of the L-p-boronophenylalanine-fructose complex in humans and derived from pharmacokinetic studies of 24 patients in Phase I clinical trials of boron neutron capture therapy. The 10B concentration profile in blood exhibits a characteristic rise during the infusion to a peak of approximately 32 microg/g (for infusion of 350 mg/kg over 90 min) followed by a biexponential disposition profile with harmonic mean half-lives of 0.32 +/- 0.08 and 8.2 +/- 2.7 h, most likely due to redistribution and primarily renal elimination, respectively. The mean model rate constants k12, k21, and k10 are (mean +/- SD) 0.0227 +/- 0.0064 min(-1), 0.0099 +/- 0.0027 min(-1), 0.0052 +/- 0.0016 min(-1), respectively, and the central compartment volume of distribution V1 is 0.235 +/- 0.042 L/kg. In anticipation of the initiation of clinical trials using an intense neutron beam with concomitantly short irradiations, the ability of this model to predict, in advance, the average blood 10B concentration during brief irradiations was simulated in a retrospective analysis of the pharmacokinetic data from these patients. The prediction error for blood boron concentration and its effect on simulated dose delivered for each irradiation field are reported for three different prediction strategies. In this simulation, error in delivered dose (or, equivalently, neutron fluence) for a given single irradiation field resulting from error in predicted blood 10B concentration was limited to less than 10%. In practice, lower dose errors can be achieved by delivering each field in two fractions (on two separate days) and by adjusting the second fraction's dose to offset error in the first. PMID- 12749711 TI - Pharmacokinetics of sodium borocaptate: a critical assessment of dosing paradigms for boron neutron capture therapy. AB - The pharmacokinetics of sodium borocaptate (BSH), a drug that has been used clinically for boron neutron capture therapy (BNCT) of malignant brain tumors, have been characterized by measuring boron concentrations by direct current plasma-atomic emission spectroscopy (DCP-AES) in a group of 23 patients with high grade gliomas. The disposition of BSH following intravenous (i.v.) infusion, which was determined by measuring plasma boron concentrations by DCP-AES, was consistent with a three-compartment open model with zero-order input and first order elimination from the central compartment. Boron disposition was linear over the dose range of 26.5-88.2 mg BSH/kg body weight (b.w.), corresponding to 15-50 mg boron/kg b.w. Mean total body boron plasma clearance was 14.4 +/- 3.5 ml/min and the harmonic mean half-lives (range) were 0.6 (0.3-3.7), 6.5 (4.8-10.1) and 77.8 (49.6-172.0) h for the alpha, beta, and gamma disposition phases, respectively. Using an empirically determined plasma: blood boron concentration ratio of 1.3 +/- 0.2, the calculated total body boron blood clearance was 18.5 +/ 4.5 ml/min. In order to develop a model for selecting the optimum dosing paradigm, a pharmacokinetic correlation was established between the boron content of normal brain, solid tumor, and infiltrating tumor to the shallow tissue pharmacokinetic compartment (C2). Based on our model, it was concluded that although multiple i.v. infusions of BSH might increase absolute tumor boron concentrations, they will not improve the tumor: plasma boron concentration ratios over those attainable by a single i.v. infusion. The results from our study are confirmatory of those previously reported by others when blood sampling has been carried out for a sufficient period of time to adequately characterize the pharmacokinetics. PMID- 12749715 TI - Identifying factors associated with disability-related differences in breast cancer screening (United States). AB - OBJECTIVE: The purpose of this study was to identify factors that could explain breast cancer underutilization among women age 40 and older with disabilities. METHODS: The data are part of the 1996 Medical Expenditure Panel Survey (MEPS), a nationally representative sample of medical care use and expenditures in the United States. Two different definitions of disability were used: limitations in activities of daily living (ADL) and limitations in instrumental activities of daily living (IADL). Annual mammography was used as the outcome measure. The data are restricted to noninstitutionalized women at least 40 years of age. RESULTS: Crude odds ratios showed that women with long-term limitations in their ADLs or IADLs were less likely to be screened for breast cancer compared to those without such limitations. These associations remained while controlling for possible confounders and were observed among women age 40 or older, those 50-69, and among women 70 years of age and older. CONCLUSIONS: Reasons for the underutilization of breast cancer screening among women with long-term disabilities remain elusive. Future studies need to examine additional factors in order to improve screening use, especially among women with long-term disabilities who are 50-69 years of age, for whom screening has been shown to be beneficial in terms of reduced risk of mortality from breast cancer. PMID- 12749714 TI - Common challenges and problems in clinical trials of boron neutron capture therapy of brain tumors. AB - Clinical trials for binary therapies, like boron neutron capture therapy (BNCT), pose a number of unique problems and challenges in design, performance, and interpretation of results. In neutron beam development, different groups use different optimization parameters, resulting in beams being considerably different from each other. The design, development, testing, execution of patient pharmacokinetics and the evaluation of results from these studies differ widely. Finally, the clinical trials involving patient treatments vary in many aspects such as their dose escalation strategies, treatment planning methodologies, and the reporting of data. The implications of these differences in the data accrued from these trials are discussed. The BNCT community needs to standardize each aspect of the design, implementation, and reporting of clinical trials so that the data can be used meaningfully. PMID- 12749713 TI - The use of positron emission tomography to develop boron neutron capture therapy treatment plans for metastatic malignant melanoma. AB - Centers in Japan and the United States are extending boron neutron capture therapy (BNCT) to the treatment of malignant melanoma (MM). Positron emission tomography (PET) has been used to image glioblastoma multiforme with 18F boronophenylalanine (18F-BPA) for the purpose of generating 10B distribution maps. These distribution maps can be used to improve the BNCT treatment planning. 18F-BPA was given to a patient with widely metastatic MM involving the thorax and brain. 18F-BPA PET scans of the chest and the head were obtained and compared to the computed tomograms (CT) and magnetic resonance (MR) images. The lung metastases seen on the chest CT images and intracranial metastases seen on CT and MR images were correlated with the PET images. The PET images clearly identified a brain lesion that was difficult to identify on MR and CT images. The 18F-BPA lung and peri-oral mucous gland activity was intense indicating a relatively high concentration of BPA. The intensity seen in the peri-oral mucous glands is consistent with the experiences in the BNCT clinical trials. These results have implications in the use of BNCT outside of the cranium. The PET images allow the generation of treatment plans that are consistent with the clinical findings. PET imaging with 18F-BPA can be used to identify potential tumors that may be amenable to BNCT and to improve treatment plans prior to BNCT. PMID- 12749716 TI - Cancer in Indigenous Australians: a review. AB - OBJECTIVES: To summarize for the first time evidence of the impact of cancer on Indigenous Australians. METHODS: Medline search of peer-reviewed scientific journals, and extensive search of reports of government agencies, publications of cancer registries and non-government organizations, and other non-peer-reviewed sources. RESULTS: Indigenous Australians have much higher incidence rates than other Australians of cancers of the lung, liver, and cervix; but much lower rates of cancers of the breast, colon and rectum, prostate, melanoma of skin, and lymphoma. Some of these differences can be explained, in part at least, by differences in risk factor prevalence. Indigenous Australians also have higher mortality and lower survival from cancer as a whole than other Australians. More advanced disease at diagnosis, and possibly poorer treatment, are partly responsible for these differences, but other factors may also be involved. CONCLUSIONS: Less accessible and less effective health programs are as great a problem for cancer control as for other aspects of Indigenous health in Australia. Major improvements in preventive services, screening, primary care, and specialist treatment services are required to reduce cancer incidence and improve cancer outcomes for Australia's Indigenous people. PMID- 12749717 TI - Case-control study of lung cancer during 1994-1997 in the birth cohort in Tasmania, Australia, with an excess of female cases during 1983-1992. AB - OBJECTIVE: In Tasmania, Australia, lung cancer incidence for 25-44-year-old women has reached that of 25-44-year-old men despite less smoking by the women. We investigated whether this could be due to greater female-than-male relative risk for smoking. METHODS: This was a case-control study of lung cancer in the 1939 1964 birth cohort. In person (n = 100) or by proxy, 158 of the 160 cases arising during 1994-1997 were interviewed. Controls were a representative sample of the cohort (response 82.8%). Detailed measurements of tobacco smoking were made by questionnaire, and using the results of 17 machine tests of cigarette "tar" yields. RESULTS: The male smokers had greater accumulated exposure to smoking and, in reversal of the previously reported excess of female cases in this cohort, most (99/160) of the 1994-1997 cases were men. Nevertheless, the proportions attributable to smoking were similar: 0.86 (0.76-0.97) of male cases, and 0.87 (0.74-0.99) of female cases. Calculated relative to male never-smokers, the estimated relative risks were similar for male and female smokers, particularly with exposure measured by cumulative tar yield of all cigarettes smoked. CONCLUSIONS: We found no compelling evidence of greater susceptibility to lung cancer for female smokers. PMID- 12749718 TI - Second primary tumors in patients with upper aerodigestive tract cancers: joint effects of smoking and alcohol (United States). AB - OBJECTIVE: This study evaluated the joint effects of tobacco smoking and alcohol consumption on the risk of second primary tumors (SPT) in patients with early stage head and neck squamous cell carcinoma (HNSCC). METHODS: Data are presented for 1181 patients enrolled in a placebo-controlled chemoprevention trial of 13 cis-retinoic acid. Nearly 17% of patients presented with a SPT. The log rank test and Cox proportional hazards model were used to examine risk factors for SPT development. RESULTS: After adjusting for the time from the index diagnosis to randomization, age at diagnosis, stage, and site of the primary cancer, the factors that emerged as simultaneous predictors of SPT development were continued smoking and alcohol intake after the index diagnosis. Increased SPT risk was associated with older age (RR = 2.1; 95% CI 1.5-2.8); stage II diagnosis (RR = 1.5; 95% CI 1.1-2.1); index diagnosis of pharyngeal cancer (RR = 1.6; 95% CI 1.1 2.5); current smoking at registration (RR = 2.1; 95% CI 1.3-3.6) and continued alcohol consumption post-diagnosis (RR = 1.3; 95% CI 1.0-1.7). CONCLUSION: Important associations exist between SPT development and continued smoking and alcohol consumption after treatment for HNSCC. PMID- 12749719 TI - Occupation and the risk of adult glioma in the United States. AB - OBJECTIVE: Previous studies have observed increased glioma incidence associated with employment in the petroleum and electrical industries, and in farming. Several other occupations have also been associated with increased risk, but with inconsistent results. We evaluated associations between occupational title and glioma incidence in adults. METHODS: Cases were 489 patients with glioma diagnosed from 1994 to 1998 at three United States hospitals. Controls were 799 patients admitted to the same hospitals for non-malignant conditions. An experienced industrial hygienist grouped occupations that were expected to have similar tasks and exposures. The risk of adult glioma was evaluated for those subjects who ever worked in an occupational group for at least six months, those who worked longer than five years in the occupation, and those with more than ten years latency since starting work in the occupation. RESULTS: Several occupational groups were associated with increased glioma incidence for having ever worked in the occupation, including butchers and meat cutters (odds ratio [OR] = 2.4; 95% confidence limits [CL]: 1.0, 6.0), computer programmers and analysts (OR = 2.0; 95% CL: 1.0, 3.8), electricians (OR = 1.8; 95% CL: 0.8, 4.1), general farmers and farmworkers (OR = 2.5; 95% CL: 1.4, 4.7), inspectors, checkers, examiners, graders, and testers (OR = 1.5; 95% CL: 0.8, 2.7), investigators, examiners, adjustors, and appraisers (OR = 1.7; 95% CL: 0.8, 3.7), physicians and physician assistants (OR = 2.4; 95% CL: 0.8, 7.2), and store managers (OR = 1.6; 95% CL: 0.8, 3.1), whereas occupation as a childcare worker was associated with decreased glioma incidence (OR = 0.4; 95% CL: 0.2, 0.9). These associations generally persisted when the subjects worked longer than five years in the occupation, and for those with more than ten years latency since starting to work in the occupation. CONCLUSIONS: This is our first analysis of occupation and will guide future exposure-specific assessments. PMID- 12749720 TI - Breast cancers among very young premenopausal women (United States). AB - OBJECTIVE: To assess risk factors for breast cancer among very young compared to older premenopausal women. METHODS: Between 1990 and 1992 a population-based case control study conducted in Atlanta, GA, Seattle/Puget Sound, WA, and central NJ interviewed 3307 premenopausal women aged 20-54 years. Logistic regression models estimated adjusted relative risks (RR) and 95% confidence intervals (CI) for each of three 10-year age groups. RESULTS: Among the youngest age group (<35 years, n = 545), significant predictors of risk included African-American race (RR = 2.66: 95% CI 1.4-4.9) and recent use of oral contraceptives (RR = 2.26; 95% CI 1.4 3.6). Although these relationships were strongest for estrogen receptor-negative (ER-) tumors (RRs of 3.30 for race and 3.56 for recent oral contraceptive use), these associations were also apparent for young women with ER+ tumors. Delayed childbearing was a risk factor for ER+ tumors among the older premenopausal women (Ptrend < 0.01), but not for women <35 years in whom early childbearing was associated with an increased risk, reflecting a short-term increase in risk immediately following a birth. Family history of early-onset breast cancer was more strongly associated with risk among women <35 years (RR = 3.22) than those 45-54 years (RR = 1.51). Risk factors for premenopausal breast cancer not significantly modified by age at diagnosis included early age at menarche, low body mass index, and heavy alcohol consumption. CONCLUSION: These findings suggest the possibility that women who develop breast cancers at very young ages may be etiologically as well as clinically distinct. PMID- 12749721 TI - Risk of prostate cancer associated with a family history in an era of rapid increase in prostate cancer diagnosis (Australia). AB - OBJECTIVE: We conducted a case-control study of prostate cancer and familial risk of the disease in Australia between 1994 and 1998, a period during which the incidence of prostate cancer increased dramatically with widespread use of prostate-specific antigen (PSA) testing. METHODS: 1475 cases and 1405 controls were asked about prostate cancer in their first-degree relatives. Odds ratios (OR) were calculated using logistic regression. RESULTS: Cases were more likely to report a family history of prostate cancer than controls (OR 3.0; 95% confidence interval (CI) 2.3-3.9) and cases reporting an affected relative were younger (58.8 versus 60.9 years, p < 0.0001). The OR for an affected first-degree relative increased with increasing number of affected relatives and decreased with increasing age of the case. The OR for more than one affected first-degree relative was 6.9 (95% CI 2.7-18). The OR for an affected brother was 3.9 (95% CI 2.5-6.1) and for an affected father was 2.9 (95% CI 2.1-3.9) but these were not significantly different (p = 0.2). When analyses were repeated including only diagnoses made in relatives prior to 1992, the risks were generally similar except that the OR for an affected brother decreased to 3.1 (95% CI 1.2-3.9). When only relatives' diagnoses made after 1991 were included results were again similar to those for all relatives, although the effect for brothers was greater and the attenuation with age at diagnosis dissipated. CONCLUSIONS: The recent introduction of PSA testing that has resulted in a greater prevalence of apparent prostate cancer, does not appear to have substantially altered familial risks of disease, although effects associated with brothers may be inflated. PMID- 12749722 TI - Cancer incidence patterns in Koreans in the US and in Kangwha, South Korea. AB - OBJECTIVE: In the US, Koreans are a rapidly growing group and comprised 10.5% of the total Asian population as of 2000. However, little has been published regarding cancer patterns in this subpopulation. METHODS: Using data from the Surveillance, Epidemiology, and End Results program, the California Cancer Registry, and the International Association for Research on Cancer, we compared age-adjusted and age-specific incidence rates for cancers of the prostate, breast, cervix, lung, colon, rectum, stomach, liver, and esophagus in US Koreans with rates of these cancers in residents of Kangwha, South Korea, and in US whites as a reference. RESULTS: While the most frequently diagnosed cancer was lung among US Korean males and breast among US Korean females, it was stomach cancer for both sexes in Kangwha. Rates of prostate, breast, and colon cancer were considerably higher for Koreans in the US than in Kangwha, but were not as high as in whites. Cervical and stomach cancers showed the opposite racial/ethnic pattern, with rates highest in Kangwha, intermediate among US Koreans, and lowest among whites. Rates of rectal cancer in females and esophageal cancer in males were two-times higher in Kangwha than in US Koreans but esophageal cancer rates were similar between US Koreans and whites. Liver cancer rates were similar between Kangwha residents and US Koreans, but nearly 10-times lower among whites. CONCLUSIONS: Although these comparisons may have methodologic limitations, including data quality and racial/ethnic misclassification, the differences seen in migrant and native Koreans for some cancers warrant further investigation in this growing subpopulation. PMID- 12749724 TI - Patterns of postmenopausal progestin use with estrogen in relation to endometrial cancer (United States). AB - OBJECTIVE: To investigate the risk of endometrial cancer associated with various regimens of postmenopausal hormone therapy. METHODS: Data from a population-based case-control study were analysed that included 591 women newly diagnosed with endometrial cancer, aged 40-79, and who were reported to Wisconsin's statewide tumor registry in 1991-1994. Similarly aged population controls (n = 2045) were randomly selected from lists of licensed drivers and Medicare beneficiaries. Information on hormone use and other factors was obtained through telephone interviews. RESULTS: Ever use of estrogen-progestin hormones was associated with increased endometrial cancer compared to women who had never used hormones; the increase per year of use was 7% (95% confidence interval 1-13%). Both cyclic (<10 days per month) and continuous progestin used with estrogen were associated with comparable twofold increases in risk relative to non-users. There was no increased risk associated with progestin used for 10-21 days per month. CONCLUSIONS: Both continuous and cyclic progestin regimens are associated with a much lower risk of endometrial cancer than estrogen alone. However, many women using these regimens remain at significantly increased risk of endometrial cancer. PMID- 12749723 TI - Building the infrastructure for nationwide cancer surveillance and control--a comparison between the National Program of Cancer Registries (NPCR) and the Surveillance, Epidemiology, and End Results (SEER) Program (United States). AB - OBJECTIVE: In preparation for jointly publishing official government cancer statistics, the Centers for Disease Control and Prevention (CDC) and the National Cancer Institute (NCI) compared incidence rates from NCI's Surveillance Epidemiology and End Results (SEER) Program and CDC's National Program of Cancer Registries (NPCR). METHODS: Data for 1999 covering 78% of the US population were obtained from SEER and selected NPCR registries that met high quality data criteria. Incidence rates (per 100,000 population) were age-adjusted to the 2000 US standard population, and 95% gamma confidence intervals were estimated. RESULTS: NPCR rates for all sites combined were higher than SEER rates (males: NPCR 553.6, SEER 538.7; females: NPCR 420.8, SEER 412.5), but rates for specific cancer sites varied by registry program. Rates for colon cancer (males: NPCR 47.0, SEER 42.7; females: NPCR 36.5, SEER 33.8) and tobacco-related cancers were higher in NPCR than SEER. In contrast, NPCR rates were lower than SEER rates for cancers of the female breast (NPCR 134.0, SEER 135.9), prostate (NPCR 162.0, SEER 170.2), and melanoma as well as for cancers more common among Asians and Pacific Islanders (e.g., stomach cancer). CONCLUSIONS: Rate differences may arise from population differences in socio-demographic characteristics, screening use, health behaviors, exposure to cancer causing agents or registry operations factors. PMID- 12749725 TI - Implications of genetic testing in the management of colorectal cancer. AB - The prognosis of patients with colorectal cancer is impacted by various factors at the time of diagnosis, including location of the tumor, gender, age and overall performance status of the patient. Optimal postoperative management of patients who have undergone successful tumor resection involves the utilization of reliable determninants of prognosis to help select patients who would benefit from adjuvant treatment, while sparing others from drug-related adverse effects. Tailoring chemotherapy for patients with disseminated cancer, or for patients who receive adjuvant chemotherapy, is also critical. Interpatient differences in tumor response and drug toxicity are common during chemotherapy. Genomic variability of key metabolic enzyme complexes, drug targets, and drug transport molecules is an important contributing factor. The identification of genetic markers of response and prognosis will aid in the development of more individualized chemotherapuetic strategies for cancer patients. Potential prognostic indicators in colorectal cancer include oncogenes, tumor suppressor genes, genes involved in angiogenic and apoptotic pathways and cell proliferation, and those encoding targets of chemotherapy. Specifically, molecular markers such as deletion of 18q (DCC), p27 and microsatellite instability are promising as indicators of good or poor prognosis. Molecular determinants of efficacy and host toxicity of the most commonly used drugs in colorectal cancer, fluoracil, irinotecan and oxaliplatin, are being investigated. Alterations in gene expression, protein expression and polymorphic variants in genes encoding thymidylate synthase, dihydropyrimidine dehydrogenase, dUTP nucleotidehydrolase and thymidine phosphorylase (for fluoropyrimidine-based chemotherapy), uridine diphosphate glucosyltransferase (UGT) 1A1 and carboxylesterase (for irinotecan therapy), and excision repair cross complementing genes (ERCC1 and ERCC2) and glutathione-S-transferase P1 (for oxalilplatin-based regimens) may be useful as markers for clinical drug response, survival and host toxicity. PMID- 12749726 TI - Genetic predisposition to neuroleptic malignant syndrome : implications for antipsychotic therapy. AB - The pathogenetic mechanism of neuroleptic malignant syndrome (NMS), a potentially lethal adverse effect of antipsychotics, is not well understood. In addition to acquired risk factors, clinical observations suggest a number of genetic factors predisposing patients to NMS. Recent findings in pharmacogenetics indicate that the genetic polymorphisms for drug-metabolizing enzymes, drug transporters, and possibly drug-targeting molecules, are associated with the interindividual differences in drug responses concerning both efficacy and adverse reactions. Genetic association studies have sought to identify polymorphisms influencing susceptibility to NMS, especially with respect to the dopamine D(2) receptor, serotonin receptor, and cytochrome p450 2D6. While a few candidate polymorphisms were associated with NMS, a large controlled study is needed to attain statistical power. On the other hand, NMS might include heterogeneous conditions with common characteristic symptoms but different causative mechanisms. Further analysis of individuals with identified genetic mutations or polymorphisms should advance our understanding of mechanisms underlying NMS. PMID- 12749727 TI - Clinical potential of respirable antisense oligonucleotides (RASONs) in asthma. AB - The human genome project, as well as advances in our understanding of asthma susceptibility, are yielding novel candidate targets for disease intervention. The normalization of up-regulated gene expression may treat or improve the disease outcome. However, only some of these gene product targets may be 'tractable', i.e. amenable to blockade by small, orally active, organic molecules. The remainder have been termed 'non-tractable'. For over a decade, antisense oligonucleotides (ASONs) have been used as tools to evaluate the importance of specific gene products in vitro. In recent years evidence has accumulated indicating their potential as a viable new therapeutic approach in their own right, being able to block 'non-tractable' targets as well as 'tractable' targets.Distribution, cell-specific uptake, and effectiveness of aerosolized phosphorothioate ASONs are currently being evaluated in animal models. The results demonstrate broad distribution throughout the lung, and uptake by all of the cell types examined to date. Functionality has been demonstrated against diverse targets, including nuclear transcription factors, tyrosine kinases, G-protein coupled receptors, cytokine receptors, growth factors, and chemokines.EPI-2010, a respirable ASON (RASON) against the adenosine A(1) receptor, is the first test case for this new class of respiratory therapeutics. The rationale for EPI-2010 is that overactivity of the adenosine signaling pathway in asthmatic lungs contributes to airway inflammation and hyperresponsiveness. EPI-2010 binds to the initiation codon of the adenosine A(1) receptor mRNA, and thereby blocks translation and targets the message for degradation by RNase. EPI-2010 is apparently metabolized locally by endogenous nucleases confining its activity to the airways. Phase I clinical trials have shown EPI-2010 to be well-tolerated, with indications of efficacy. In conclusion, one important application of RASONs is in addressing up-regulated disease targets, only some of which are 'tractable' by small molecules. It is hoped that this will yield new therapeutic options to the benefit of patients with asthma and allergic disorders. PMID- 12749728 TI - Proteomic prediction of disease outcome in cancer : clinical framework and current status. AB - Better than gene sequencing or quantitative amplification, proteomics tools allow the study of tumor phenotype. Indeed, most current prognostic tests in cancer (carcinoembryonary antigen [CEA], prostate-specific antigen [PSA], CA 19-1, CA 125, alpha-fetoprotein [AFP], etc.) are based on the detection and quantification of single proteins in body fluids. However, a common characteristic of these tests is their relatively low predictive value, so that they are usually complemented with other procedures such as biopsy and/or endoscopy. Recently, improved analytical and bioinformatics tools have driven the attention on pattern recognition approaches rather then single-marker tests for prognostic forecasting. It is expected that predicting metastasization on the basis of tumoral protein patterns will soon be a reality. However, currently available technologies either limit the number of proteins that can be analyzed simultaneously or they are expensive, difficult, and time-consuming. Moreover, the tools adapted for expression proteomics might not be the same as those for prognostic studies that require investigation of protein function over time. We believe that clinical proteomics research designed within a precise clinical and pathology framework should be strongly supported, since many prognostic factors are determined not by the tumor itself, but by the patient, the treatment and the environment. PMID- 12749729 TI - Utilizing functional genomics to identify new pain treatments : the example of nociceptin. AB - Nociceptin/orphanin FQ (noc/oFQ) is the first novel bioactive substance to have been discovered by the implementation of a functional genomics/reverse pharmacology approach. The neuropeptide was indeed identified in brain extracts as the natural ligand of a previously cloned orphan G protein-coupled receptor, the opioid receptor-like 1 (ORL1) receptor. Since its discovery in 1995, noc/oFQ has been the subject of intensive study to establish its role in normal brain function and its possible involvement in neurophysiopathology. Although the neuropeptide, an inhibitor of neuronal activity, has been found to have a wide spectrum of pharmacological effects in vivo, none has been as intensively investigated as its action on nociception and nociceptive processing. There is now substantial evidence that noc/oFQ has a modulatory role in nociception. However, dependent on the dose and site of injection, and possibly the animal's genetic background and even psychological status, the peptide has been variously reported to cause allodynia, hyperalgesia, analgesia, and even pain, in rodents. Overall, noc/oFQ tends to facilitate pain when administered supraspinally, and to inhibit it when administered spinally. These opposing effects beg the obvious, yet still unanswered, question as to what would be the net effect on nociception of an ORL1 receptor ligand, agonist or antagonist, able to target supraspinal and spinal sites simultaneously. Owing to the research effort of several drug companies, such ligands, i.e. nonpeptidic, brain-penetrating agonists and antagonists, have recently been produced whose systematic screening in animal models of acute and inflammatory pain may help validate the ORL1 receptor as the target for novel, non-opioid analgesics. PMID- 12749730 TI - Engineered catalytic RNA and DNA : new biochemical tools for drug discovery and design. AB - Since the fundamental discovery that RNA catalyzes critical biological reactions, the conceptual and practical utility of nucleic acid catalysts as molecular therapeutic and diagnostic agents continually develops. RNA and DNA catalysts are particularly attractive tools for drug discovery and design due to their relative ease of synthesis and tractable rational design features. Such catalysts can intervene in cellular or viral gene expression by effectively destroying virtually any target RNA, repairing messenger RNAs derived from mutant genes, or directly disrupting target genes. Consequently, catalytic nucleic acids are apt tools for dissecting gene function and for effecting gene pharmacogenomic strategies. It is in this capacity that RNA and DNA catalysts have been most widely utilized to affect gene expression of medically relevant targets associated with various disease states, where a number of such catalysts are presently being evaluated in clinical trials. Additionally, biotechnological prospects for catalytic nucleic acids are seemingly unlimited. Controllable nucleic acid catalysts, termed allosteric ribozymes or deoxyribozymes, form the basis of effector or ligand-dependent molecular switches and sensors. Allosteric nucleic acid catalysts promise to be useful tools for detecting and scrutinizing the function of specified components of the metabolome, proteome, transcriptome, and genome. The remarkable versatility of nucleic acid catalysis is thus the fountainhead for wide-ranging applications of ribozymes and deoxyribozymes in biomedical and biotechnological research. PMID- 12749731 TI - Serotonin transporter promoter polymorphism in African Americans : allele frequencies and implications for treatment. AB - BACKGROUND: Americans of African ancestry are less likely to receive a selective serotonin reuptake inhibitor (SSRI) for treatment of major depressive disorder than Americans of European ancestry. A functional insertion/deletion polymorphism in the promoter of the serotonin transporter (5-HTT) gene SLC6A4 has been shown to modulate SLC6A4 transcription, affecting response to SSRIs. Several studies in populations of predominantly European ancestry have consistently found that the SLC6A4 promoter polymorphism (referred to as the 5-HTT-linked polymorphic region; 5-HTTLPR) long (L) allele is associated with better response to SSRI treatment than the short (S) allele. OBJECTIVE: The frequency of SLC6A4 (5-HTTLPR) alleles in 865 black Americans and Afro-Caribbeans was examined to assess possible implications for treatment. STUDY DESIGN AND METHODS: SLC6A4 (5-HTTLPR) genotypes were determined in individuals with self-identified African ancestry from South Carolina (n = 489), western Pennsylvania (n = 207), and Tobago (n = 169). Frequencies were compared using chi-square analyses. RESULTS: It was verified that the L allele is highly prevalent in Americans of African ancestry, ranging from 77% in western Pennsylvania to 87% in South Carolina. The frequency of the SLC6A4-(L) allele is significantly higher in African-Americans than has been reported for European-Americans (typically 56-60%). There are both statistically significant geographic differences and slight deviations from Hardy-Weinberg equilibrium. CONCLUSIONS: Given the potential influence on treatment response, these findings have implications for the use of SSRIs in this population. The results suggest that additional studies to examine the impact of these alleles on treatment response in African-Americans are warranted. PMID- 12749732 TI - Is there a role for adjuvant high-dose interferon-alpha-2b in the management of melanoma? AB - The knowledge that melanoma is susceptible to attack by the host's immune system has resulted in the testing of a variety of immunotherapies. Interferon-alpha-2b, which has several anti-tumour mechanisms including an antiproliferative effect, an anti-angiogenesis effect, the enhancement of natural-killer cell activity and the upregulation of tumour antigen presentation, has shown tremendous potential. Early trials using low-dose and intermediate-dose regimens demonstrated no benefit to survival. However, the Eastern Cooperative Oncology Group trial EST 1684, showed that a high-dose regimen involving an induction phase of intravenous interferon-alpha-2b 20 MU/m(2) 5 days a week for 4 weeks, followed by a maintenance phase of subcutaneous 10 MU/m(2) 3 days a week for the remainder of a year, led to significant improvements in both disease-free and overall survival compared with observation. On the basis of these results, the US FDA approved high-dose interferon-alpha-2b for the post-surgical adjuvant therapy of high-risk melanoma. Unfortunately, the results of subsequent trials involving high-dose interferon-alpha-2b have not been as clear, and its role in the adjuvant treatment of melanoma remains controversial. Concerns remain regarding the design and interpretation of the clinical trials, the cost and toxicity of treatment, and the appropriate selection of patients who should be treated. This article reviews the existing data and attempt to address the arguments for and against a role for adjuvant high-dose interferon-alpha-2b in the management of melanoma. PMID- 12749733 TI - Management of patients with recurrent vulvovaginal candidiasis. AB - Recurrent vulvovaginal candidiasis (RVVC) is by no means uncommon and is a source of considerable physical discomfort in addition to serving as a major therapeutic challenge. The syndrome is multifactorial in aetiology and hence management strategies must recognise the complex aetiological pathways. Many women receiving the misplaced diagnosis of RVVC have a variety of other infectious and non infectious entities presenting with identical symptoms. Hence the first step in management is confirming the diagnosis of RVVC including microbial confirmation and species identification. Efforts should be made to identify and correct a causal mechanism. Maintenance suppressive azole antifungal regimens are highly effective in controlling symptoms, although cure is less common. Further advances in achieving higher cure rates await the availability of non-azole fungicidal agents. PMID- 12749738 TI - Tramadol/paracetamol. AB - The orally administered fixed combination tablet of tramadol (centrally-acting opiate) plus paracetamol (acetaminophen; nonopiate, nonsalicylate analgesic) [37.5/325 mg] provides effective analgesia in patients with moderate to severe acute pain and those with chronic painful conditions characterised by intermittent exacerbations of pain. Two tramadol/paracetamol 37.5/325 mg tablets provided greater relief of dental pain over an 8-hour period than either agent alone, with a faster onset of action than tramadol alone and a longer duration of action than either agent as monotherapy. In patients with postoperative dental pain, two tramadol/paracetamol tablets (37.5/325 mg) provided similar analgesia to hydrocodone/paracetamol 10/650 mg over an 8-hour period. The addition of one or two tramadol/paracetamol 37.5/32 5mg tablets (up to four times daily) for 5 days to existing NSAID or cyclo-oxygenase-2 inhibitor analgesic therapy provided effective pain relief in patients with osteoarthritis flare pain. Tramadol/paracetamol 37.5/325 mg provided similar efficacy to that of codeine/paracetamol 30/300 mg in patients with chronic back pain in a 4-week, randomised, double-blind trial (a maximum of 10 tablets or capsules per day of the active drug). PMID- 12749734 TI - Darbepoetin alfa: in patients with chemotherapy-related anaemia. AB - Darbepoetin alfa, novel erythropoiesis stimulating protein closely related to human erythropoietin, has been developed for the treatment of chemotherapy related anaemia in patients with non-myeloid malignancies. In three 12-week, phase II studies in patients with cancer and chemotherapy-related anaemia, subcutaneous darbepoetin alfa, administered in once-weekly or 2-, 3- or 4-weekly regimens, dose-dependently increased the mean haemoglobin levels. In a randomised, double-blind, phase III study in 320 patients with lung cancer and chemotherapy-related anaemia, recipients of subcutaneous darbepoetin alfa 2.25 micro g/kg once weekly, received red blood cell (RBC) transfusion approximate, equals 2-fold less frequently than placebo recipients (p < 0.001). In the same study, patients receiving darbepoetin alfa also received fewer standard units of RBC for transfusion and had greater haematopoietic response rate than placebo recipients (both p < 0.001). Subcutaneous darbepoetin alfa 2.25 micro g/kg once weekly also reduced patient-reported fatigue (assessed by a quality-of-life questionnaire) [p = 0.019 vs placebo]. black triangle Darbepoetin alfa was generally well tolerated in clinical trials. The most frequent darbepoetin alfa related adverse events were: body oedema, arthralgia and skin rash. PMID- 12749741 TI - Lamivudine/zidovudine/abacavir: triple combination tablet. AB - The triple combination tablet containing lamivudine (150 mg), zidovudine (300 mg) and abacavir (300 mg, as abacavir sulfate) is a new formulation of three nucleoside analogue reverse transcriptase inhibitors. Two studies in treatment naive patients (one double-blind, one nonblind) have reported that lamivudine/zidovudine (dual combination tablet) plus abacavir showed efficacy similar to that of lamivudine/zidovudine plus indinavir. In both studies, similar numbers of patients in each treatment group had plasma HIV RNA levels 400 copies/mL, was reported in 22% of patients in both treatment groups at week 48. Treatment-naive patients receiving lamivudine/zidovudine/abacavir combination therapy experienced several adverse events, including nausea, malaise/fatigue and vomiting. PMID- 12749744 TI - Tegaserod: a review of its use in the management of irritable bowel syndrome with constipation in women. AB - The treatment of irritable bowel syndrome with constipation (IBS-C) has historically been based on the severity of symptoms, with education, reassurance, dietary advice, bulking agents and laxative therapy offered as appropriate. Tegaserod (Zelnorm, Zelmac) is the first selective serotonin 5-HT(4) receptor partial agonist to be approved for the treatment of this syndrome. Tegaserod is active against multiple irritable bowel syndrome (IBS) symptoms; it stimulates gut motility and reduces visceral sensitivity and pain. The drug does not cure IBS and was not designed to treat the diarrhoea-predominant version. Its efficacy in men has not been established. Three large well designed clinical trials of tegaserod 6 mg twice daily for 12 weeks in patients (mainly women) with IBS-C have demonstrated superiority versus placebo in global relief from symptoms. Global relief response rates were 38.4-46.8% with tegaserod 6 mg twice daily and 28.3-38.8% with placebo (p < 0.05-0.0001 vs placebo). The relative increases in response rates with tegaserod 6 mg twice daily over the already high responses in the placebo groups ranged from 12-65% after 4-12 weeks of treatment. A response was seen within the first week. The proportion of patients with satisfactory relief from symptoms fell over the 4-week period following withdrawal of tegaserod and placebo, but did not reach baseline levels during this time. Diarrhoea has been associated with tegaserod in clinical trials (an incidence of about 10% versus 5% with placebo, usually occurring in the first week of treatment), but the drug is otherwise well tolerated. There were no apparent changes in the tolerability profile with extended tegaserod treatment (Cys), were capable of mediating methotrexate uptake above the low level in MtxRIIOuaR2-4 cells. We found that by treating the transfected cells with the small, water-soluble, thiol-reactive anionic reagent, sodium (2-sulphonatoethyl) methanethiosulphonate, methotrexate transport by several of the cysteine substituted hRFC mutants was significantly inhibited, including Q40C, G44C, E45C and I48C. Sodium (2-sulphonatoethyl) methanethiosulphonate transport inhibition of the Q40C, G44C and I48C mutants was protected by leucovorin [(6R, S)-5 formyltetrahydrofolate], indicating that these residues lie at or near a substrate-binding site. Using surface-labelling reagents [N-biotinylaminoethyl methanethiosulphonate and 3-(N-maleimidylpropionyl)biocytin, combined with 4 acetamido-4'-maleimidylstilbene-2,2'-disulphonic acid] with cysteine mutants from positions 37-48, the extracellular TMD1 boundary was found to lie between residues 39 and 40, and amino acids 44-46 and 48 were localized to the TMD1 exofacial loop. Collectively, our results imply that amino acids 40, 44, 48 and, possibly, 42 serve important roles in hRFC transport, albeit not as structural components of the putative transmembrane channel for folate substrates. PMID- 12749770 TI - Effects of p-nonylphenol and resveratrol on body and organ weight and in vivo fertility of outbred CD-1 mice. AB - The aim of this study was to analyse the multigenerational effects of para nonylphenol (NP) and resveratrol (RES) on the body weight, organ weight and reproductive fitness of outbred CD-1 mice. The data indicate that in male mice, NP had an effect on the weight of selected reproductive organs and the kidneys in the parental (P) generation males. Effects on selected reproductive organs, the liver and kidneys in the F1-generation males were also seen. In females, effects of NP on body weight and kidney weight were seen in the P generation, but no effects on any measured parameter were seen in the F1 generation. RES had no effect on body weight but did have some effect on selected male and female reproductive organs in the P generation. RES altered the spleen and liver weights of P-generation males and the kidney weight of F1-generation males. Acrosomal integrity (using a monoclonal antibody against intra-acrosomal sperm proteins) was assessed for both generations of NP- and RES-treated mice. A significant reduction in acrosomal integrity was seen in both generations of NP-treated, but not in RES-treated, mice. Fewer offspring were observed in the second litter of the F2 generation of mice treated with NP; no similar effect was seen in RES treated mice. The litter sex ratio was not different from controls. Unlike RES, NP had a negative effect on spermatogenesis and sperm quality with a resultant impact on in vivo fertility. PMID- 12749772 TI - Differential effects of hydroxamate inhibitors on PMA and ligand-induced L Selectin down-modulation: role of membrane proximal and cytoplasmic domains. AB - L-selectin is down-modulated from the cell surface in response to leukocyte activation or cross-linking with ligand mimetics such as anti-L-selectin antibodies or sulfatides. The down-modulation induced upon cellular activation with PMA is due to proteolytic shedding mediated by a cysteine metalloprotease, presumably the TNF-alpha converting enzyme (TALE), and is susceptible to inhibitors of the hydroxamate class. To determine if cross-linking-induced down modulation of L-selectin is similarly affected by these inhibitors, we exposed Jurkat T cells to the anti-L-selectin antibody Dreg 200 or sulfatides in the presence of the hydroxamate TNF-alpha protease inhibitor (TAPI). TAPI completely inhibited PMA-induced downmodulation but had no effect on that induced by the anti-L-selectin antibody or sulfatides. The downmodulation induced by an anti-TCR antibody (WT31) was only partially inhibited by TAPI. An L-selectin mutant lacking the putative TACE cleavage site and resistant to PMA-induced shedding (321del.9) was expressed in L1.2 pre-B cells. Like WT L-selectin, this mutant was down-modulated when exposed to sulfatides, or Dreg 200 and this down-modulation was unaffected by TAPI. An L-selectin cytoplasmic tail deletion mutant (344del.15) expressed in L1.2 pre-B cells was down-modulated by PMA or sulfatides, but not Dreg 200. Electrophoretic analysis of L-selectin immunoprecipitated from the supernatants of Jurkat cells treated with either sulfatides or D200 revealed a proteolytic fragment of the same size as that released from the cell surface in response to PMA. Our data indicate that the down regulation of L-selectin in response to cross-linking by ligands or TCR engagement may be mechanistically distinct from that induced by PMA. Furthermore, our results with the 344del.15 mutant suggest that the down-modulation of Lselectin induced by certain sulfated carbohydrate ligands may be initiated through surface receptors other than L-selectin itself. The abbreviations used in this paper are: TAPI, TNF-alpha protease inhibitor; TACE, TNF-alpha converting enzyme; PMA, 4(3-phorbol 12-myristate 13-acetate. PMID- 12749763 TI - The role of serine/threonine protein phosphatases in exocytosis. AB - Modulation of exocytosis is integral to the regulation of cellular signalling, and a variety of disorders (such as epilepsy, hypertension, diabetes and asthma) are closely associated with pathological modulation of exocytosis. Emerging evidence points to protein phosphatases as key regulators of exocytosis in many cells and, therefore, as potential targets for the design of novel therapies to treat these diseases. Diverse yet exquisite regulatory mechanisms have evolved to direct the specificity of these enzymes in controlling particular cell processes, and functionally driven studies have demonstrated differential regulation of exocytosis by individual protein phosphatases. This Review discusses the evidence for the regulation of exocytosis by protein phosphatases in three major secretory systems, (1) mast cells, in which the regulation of exocytosis of inflammatory mediators plays a major role in the respiratory response to antigens, (2) insulin secreting cells in which regulation of exocytosis is essential for metabolic control, and (3) neurons, in which regulation of exocytosis is perhaps the most complex and is essential for effective neurotransmission. PMID- 12749773 TI - Oligopeptide fragments of the enkephalin molecule interfere with hematopoietic cell colony formation. AB - Dipeptide Tyr-Gly and tripeptide Tyr-Gly-Gly representing the NH(2)-end of the Met-enkephalin molecule inhibited the GM-colony formation in clonal cultures of mouse bone marrow cells. Intermediate or C00H-terminal dipeptides Gly-Gly and Phe Met respectively were inneffective. The suppressive effects were not abolished by opioid receptor blocking agent naloxone and only partly so by depletion of the accessory (adherent) cells. The results are congruent with idea that neuropeptides and products of their enzymatic degradation participate in the regulation of hematopoiesis. PMID- 12749771 TI - Genome wide identification of regulatory motifs in Bacillus subtilis. AB - BACKGROUND: To explain the vastly different phenotypes exhibited by the same organism under different conditions, it is essential that we understand how the organism's genes are coordinately regulated. While there are many excellent tools for predicting sequences encoding proteins or RNA genes, few algorithms exist to predict regulatory sequences on a genome wide scale with no prior information. RESULTS: To identify motifs involved in the control of transcription, an algorithm was developed that searches upstream of operons for improbably frequent dimers. The algorithm was applied to the B. subtilis genome, which is predicted to encode for approximately 200 DNA binding proteins. The dimers found to be over represented could be clustered into 317 distinct groups, each thought to represent a class of motifs uniquely recognized by some transcription factor. For each cluster of dimers, a representative weight matrix was derived and scored over the regions upstream of the operons to predict the sites recognized by the cluster's factor, and a putative regulon of the operons immediately downstream of the sites was inferred. The distribution in number of operons per predicted regulon is comparable to that for well characterized transcription factors. The most highly over-represented dimers matched sigmaA, the T-box, and sigmaW sites. We have evidence to suggest that at least 52 of our clusters of dimers represent actual regulatory motifs, based on the groups' weight matrix matches to experimentally characterized sites, the functional similarity of the component operons of the groups' regulons, and the positional biases of the weight matrix matches. All predictions are assigned a significance value, and thresholds are set to avoid false positives. Where possible, we examine our false negatives, drawing examples from known regulatory motifs and regulons inferred from RNA expression data. CONCLUSIONS: We have demonstrated that in the case of B. subtilis our algorithm allows for the genome wide identification of regulatory sites. As well as recovering known sites, we predict new sites of yet uncharacterized factors. Results can be viewed at http://www.physics.rockefeller.edu/~mwangi/. PMID- 12749775 TI - Usefulness of (13)C-urea breath test in the diagnosis of gastric helicobacter pylori infection. AB - Helicobacter pylori chronically infects half of the human population and is associated with gastritis, peptic ulcer and gastric cancer. (13)C-urea breath test (UBT) is the main in vivo tool for the diagnosis of H. pylori infection. In this study, the safety and the accuracy of UBT were evaluated. A group of 492 dyspeptic patients was studied by UBT, the results were expressed as the difference over baseline at 30 min (DOB30). All patients were evaluated for systemic, gastrointestinal or allergic-type adverse reactions after ingestion of 75 mg (13)C-urea and citric acid in aqueous solution. The first 256 patients enrolled also underwent endoscopy and gastric biopsy. Patients positive on histology were considered infected. UBT was well tolerated and none of the 492 patients had any systemic or allergic-type adverse reaction. Among the 256 patients studied with histology, 116 were H. pylori positive on biopsies. Using 4 %o as cut-off value for DOB30,115 out of the 256 patients were positive on UBT, with only 2 false positive and 3 false negative. With this threshold, the sensitivity, specificity, and accuracy of the UBT were 97.4%, 98.5%, and 98.0%, respectively. (13)C-UBT has proven to be a safe and simple, yet accurate, test for the non-invasive diagnosis and monitoring of H. pylori infection. PMID- 12749774 TI - Trichinella spiralis infection is mediated by MCP-1 and MIP-2, while echinococcus granulosus is strongly mediated by MCP-1, but not MIP-2. AB - The chemokine supergene family are small proteins (8-10 KD) inducible in a number of pathophysiological processes. C-X-C family members are specific largely on neutrophils; whereas C-C chemokines act primarily on monocytes, T-cells, basophils and eosinophils. A wide variety of cell types produce chemokines in response to stimuli, including: infections, mitogens and inflammatory cytokines. Here we found that parasitic diseases, such as Trichinella spiralis and Echinococcus granulosus produce, MCP-1 (beta-chemokine) and MIP-2 (alpha chemokine), or MCP-1, respectively. The specificity of production of these chemokines in the two parasitic diseases may help to achieve therapeutic strategies in inhibiting inflammation. PMID- 12749777 TI - Nasal mucosal involvement in systemic lupus erythematosus: histopathologic and immunopathologic study. AB - The aim of this study was to investigate the pattern of nasal mucosal involvement in patients with Systemic Lupus Erythematosus (SLE). The authors selected patients affected by SLE with a symptomatology based on bad nasal breathing, in absence of anatomical deformities of the nose. Specimens representing eighteen histological sections of nasal mucosa were examined under the light microscope to establish a set of histopathologic and immunophahologic features. A number of significant alterations were identified. The authors indicate the possible physiopathogenetic relationship between nasal mucosal involvement and systemic disease. PMID- 12749776 TI - Suppression of adjuvant arthritis in rats with chronic bromocriptine treatment does not prevent associated oxidative stress. AB - We studied the effect of prolactin (PRL) inhibition by bromocriptine (BRC) in the first phase of adjuvant induced arthritis (AA), up to day 11(BRCl-AA), and in the whole time course of AA, up to day 23 (BRC-AA), on the development of the disease in male Lewis rats. On day 24, arthritic rats showed inhibition of PRL secretion, but not PRL mRNA expression in adenopituitaries. BRC treatment suppressed PRL serum levels and PRL mRNA expression in adenopituitaries. In BRC/-AA group PRL levels and PRLmRNA were at the level of rats with AA. Serum corticosterone (CORT) was stimulated by AA from 16.9+/-5.8 to 59.1+/-8.7 ngml(-1), p<0.05, to the same level in BRC-control (BRC-C) and BRC-AA group and further potentiated in BRCI-AA group (148.2+/-33.1 ngml(-1), p<0.05 vs. group with AA). Hind paw swelling was reduced but not completely inhibited in BRC1-AA group and totally prevented in BRC-AA rats as was the core temperature (36.5+/-0.1 degrees C vs. 37.4+/-.0.1 degrees C in AA rats on day 23, p<0.01). Serum concentration of NO-ZNO-3 rose in rats with AA to 28.7+/-2.5 &mgr;mo1L-1 against. 13.9+/-1.9 &mgr;molL(-1) in controls (p<0.01), remained elevated in BRC-AA group and was potentiated in BRC1 AA group (48.2+/-3.5 &mgr;mol(-1), p<0.01 vs. AA or BRC1-AA group) Thiobarbituric acid reactive substances (TBARS) and antioxidant capacity in the spleen were enhanced in rats with AA and to the same extent in BRC-AA or BRC1-AA groups. These results show a discrepancy between the suppression of clinical symptoms and persisting oxidative stress in AA rats after the BRC induced PRL inhibition. The potentiation of nintric oxide (NO-) production after the sudden cessation of PRL inhibition during the disease may promote further joint damage. PMID- 12749778 TI - Contact dermatoses in metal workers. AB - We studied 150 metal workers occupationally exposed to metals and metalworking fluids (MWFs) to determine the prevalence and nature of contact dermatitis. 150 office workers were used as non-exposed control group. Questionnaires were administered to evaluate occupational and non-occupational exposure. All subjects underwent a dermatological examination and patch-testing with standard allergen series and MWFs used in the plant. Twenty-eight metal workers (18.6%) presented minor skin disorders involving the hands (vs. only 2% of the controls), ten (6.6%) had major disorders (similar to the figure for the control group - 5.4%), and 112 (74.8%) had no lesions, as opposed to 92.6% of the control group. Positive patch tests were found in ten metal workers: eight had major skin disorders (six to nickel, cobalt and chromium, one to nickel and cobalt, one to nickel) and the remaining two were asymptomatic (one positive for nickel and chromium, one for nickel). Among the controls there were three cases of positivity, all among asymptomatic subjects. Patch tests with MWFs were negative. The prevalence of dermatoses among the metal workers was significantly higher than that of controls (p<0.01), and all cases of allergy in this group were provoked by metals themselves. PMID- 12749779 TI - Carbamazepine-induced hypersensitivity syndrome in a child with epilepsy. AB - Carbamazepine is an effective anticonvulsant and is considered the drug of first choice for the treatment of partial and secondarily generalized seizures. Although carbamazepine is well tolerated, many side effects have been reported in the literature. The majority of these adverse effects are transient and do not lead to the discontinuation of the therapy. We present a case of a female child, aged 11 years and 6 months, who showed an anticonvulsant hypersensitivity syndrome induced by carbamazepine. This syndrome is a rare, potentially life threatening adverse drug reaction. The patient developed a cutaneous nonpruritic rash, associated with high fever, diffuse lymphadenopathy, and arthralgias on the knees and the ankles with local signs of arthritis. Laboratory examination showed a lymphocytosis, mild thrombocytopenia, marked eosinophilia, and high transaminases. Corticosteroid therapy (betametasone 0,5 mg x 3 day) was started and carbamazepine was gradually withdrawn changing to valproic acid, with complete control of the seizures. The fever and the rash reduced gradually, beginning from the face and then disappearing completely after 10 days. Laboratory results showed a clear improvement: after 7 days the patient showed a complete normalization of the above parameters, except for transaminases. The complete normalization of these enzymes was observed after 2 weeks from the disappearance of the skin rash. PMID- 12749780 TI - Morning sudden cardiac death. AB - A circadian variation of cardiac function with peak in the early morning was documented about twenty years ago. A circadian rhythm of platelet aggregability, in the same time of the day, was demonstrated in healthy young male subjects. The morning hours were also reported as crucial for sympathetic nervous system activity, for heart rate variability, and for the abrupt rise in blood pressure. Altogether, these trigger factors may explain the high incidence of sudden cardiac death during the morning. In the primary prevention of sudden death in patients with high cardiovascular risk, many strategies were proposed, such as implantable cardioverter-defibrillators, antiarrhythmic and antihypertensive therapies, particularly beta-blockers and more recently, aspirin. Also in subjects without cardiovascular risk factors, it is predictable that early and continuous administration of low-dose aspirin, by inhibiting platelet aggregation and thrombin formation, particularly in morning hours, may represent an effective therapy for the prevention of myocardial infarction and morning sudden cardiac death. PMID- 12749781 TI - Bridging peripheral nerve defect with chitosan-collagen film. AB - OBJECTIVE: To seek new method for the treatment of peripheral nerve injury. METHODS: In rat model with sciatic nerve defect, chitosan-collagen film was sutured into conduit to bridge 5 mm, 10 mm nerve defects. Rats that underwent end to-end anastomosis were taken as controls. General observation, electrophysiological study, histological study and image analysis were performed at 4, 8, 12 weeks postoperatively. RESULTS: In 5 mm nerve defects, the quality of nerve regeneration was similar to that of the control group. For 10 mm nerve defect, nerve regeneration was inferior to that of the control group. Chitosan collagen film obviously degraded at 12 weeks postoperatively. CONCLUSIONS: Chitosan-collagen film conduit can be used to bridge peripheral nerve defect. PMID- 12749783 TI - Changes of arginine vasopressin in elderly patients with acute traumatic cerebral injury. AB - OBJECTIVE: To investigate the changes and clinical significance of arginine vasopressin (AVP) in elderly patients with acute traumatic cerebral injury. METHODS: With radioimmunoassay, the plasma levels of AVP were measured in 32 elderly patients with acute traumatic cerebral injury, 30 traumatic patients without cerebral injury and 30 healthy elderly volunteers, respectively. RESULTS: The plasma level of AVP in patients with acute traumatic cerebral injury in the early stage (48.30 ng/L +/- 8.28 ng/L) was much higher than that of the traumatic patients without cerebral injury (25.56 ng/L +/- 4.64 ng/L, P<0.01), which was much higher than that of the healthy volunteers (5.06 ng/L +/- 4.12 ng/L, P<0.01). The level of AVP in the patients with acute traumatic cerebral injury was negatively related with GCS scores. CONCLUSIONS: AVP may play an important role in the pathophysiological process in patients with acute traumatic cerebral injury in the early stage. The severer the cerebral injury is, the higher the level of AVP is, which indicates that the level of AVP may be one of the severity indices of traumatic cerebral injury in elderly patients. PMID- 12749782 TI - Relationship between expression of Bax and Bcl-2 proteins and apoptosis in radiation compound wound healing of rats. AB - OBJECTIVE: To study the relationship between the expression of Bax, Bcl-2 proteins, and apoptosis in radiation compound wound healing of rats. METHODS: Apoptosis, Bax and Bcl-2 proteins were estimated by in situ terminal labeling (TUNEL) and immunohistochemical methods. RESULTS: (1) Changes of the apoptosis in wound healing showed three typical characteristics: early occurrence, high frequency and delayed disappearance after radiation to rats when compared with those of simple wound group, which might be an important reason for radiation induced delayed wound healing. (2) The expression of Bax protein increased evidently with the increment of apoptosis and showed a good corresponding relationship with the apoptotic frequency in the process of wound healing. While the expression of Bcl-2 protein decreased obviously as the apoptosis reached a maximum and showed increasing tendency up to normal level when the apoptosis decreased distinctively. CONCLUSIONS: Bax and Bcl-2 proteins play an important role in the apoptotic regulation of radiation compound wound healing in rats. PMID- 12749784 TI - Recurrence of chronic subdural hematoma after trepanation and drainage. AB - OBJECTIVE: To sum up the causes of recurrence of chronic subdural hematoma (CSDH) from failure of trepanation and drainage and explore its prevention and treatment. METHODS: From October 1988 to June 2002 a total of 358 patients with CSDH were treated with trepanation and drainage in our hospital. Among them 15 patients had recurrence of CSDH after operation. The data of the 15 patients were reviewed retrospectively. RESULTS: Of the 15 patients, 13 were cured by retrepanation and redrainage, one cured by removal of hematoma by craniotomy with bone flap, and one, a 1-year old child, gave up reoperation due to severe encephalatrophy. CONCLUSIONS: Most CSDHs which recur after trepanation and drainage can be cured by retrepanation and redrainage. For the patients with repeated recurrence of CSDH removal of hematoma capsule can be considered. The causes of recurrence of CSDH are related to disease course, the thickness of hematoma capsule, the severity of encephalatraphy and whether the hematoma cavity is drained or irrigated completely, and operation methods. PMID- 12749785 TI - Long-term result of guided nerve regeneration with an inert microporous polytetrafluoroethylene conduit. AB - OBJECTIVE: To evaluate the long-term outcome of Polytetrafluoroethylene (PTFE) conduit in nerve repair and to provide more evidence in view of its potential application to achieve a satisfactory functional recovery in clinical settings. METHODS: Thirty-six Wistar rats had their right sciatic nerve transected and were repaired with either conventional microsuture technique (Control group, n=18) or a PTFE conduit with a gap of 5 mm left between the nerve stumps (PTFE group, n=18). At 6 and 9 months after the operation, electrophysiological assessment and measurement of gastrocnemius muscle weight were conducted and morphology of the regenerated nerves were studied with image analysis. RESULTS: At 6 months postoperatively, the nerve conduction velocity recovered to 60.86% and 54.36% (P<0.05), and the gastrocnemius muscle weight recovered to 50.89% and 46.11% (P>0.05) in the Control group and the PTFE group respectively. At 9 months postoperatively, the recovery rate was 65.99% and 58.79% for NCV (P>0.05), and 52.56% and 47.89% for gastrocnemius muscle weight (P>0.05) in the Control group and the PTFE group respectively. Regenerated nerve fibers in the PTFE group had a regular round shape with no fragmentation, wrinkling or splitting of the myelin sheath. Image analysis revealed that the ratio of the myelin area to the total fiber area was larger at 9 months than at 6 months in both groups (P<0.01). CONCLUSIONS: Microporous PTFE conduit may be an alternative for nerve repair allowing of guided nerve regeneration and functional recovery with no obvious adverse effect at long-term. PMID- 12749786 TI - Study on changes of partial pressure of brain tissue oxygen and brain temperature in acute phase of severe head injury during mild hypothermia therapy. AB - OBJECTIVE: To study the changes of partial pressure of brain tissue oxygen (PbtO2) and brain temperature in acute phase of severe head injury during mild hypothermia therapy and the clinical significance. METHODS: One hundred and sixteen patients with severe head injury were selected and divided into a mild hypothermia group (n=58), and a control group (n=58) according to odd and even numbers of hospitalization. While mild hypothermia therapy was performed PbtO2 and brain temperature were monitored for 1-7 days (mean=86 hours), simultaneously, the intracranial pressure, rectum temperature, cerebral perfusion pressure, PaO2 and PaCO2 were also monitored. The patients were followed up for 6 months and the prognosis was evaluated with GOS (Glasgow outcome scale). RESULTS: The mean value of PbtO2 within 24 hour monitoring in the 116 patients was 13.7 mm Hg +/- 4.94 mm Hg, lower than the normal value (16 mm Hg +/- 40 mm Hg ) The time of PbtO2 recovering to the normal value in the mild hypothermia group was shortened by 10 +/- 4.15 hours compared with the control group (P<0.05). The survival rate of the mild hypothermia group was 60.43%, higher than that of the control group (46.55%). After the recovery of the brain temperature, PbtO2 increased with the rise of the brain temperature. CONCLUSIONS: Mild hypothermia can improve the survival rate of severe head injury. The technique of monitoring PbtO2 and the brain temperature is safe and reliable, and has important clinical significance in judging disease condition and instructing clinical therapy. PMID- 12749787 TI - Changes of pulmonary intercellular adhesion molecule-1 and CD11b/CD18 in peripheral polymorphonuclear neutrophils and their significance at the early stage of burns. AB - OBJECTIVE: To investigate the role of intercellular adhesion molecule-1 (ICAM-1) in the accumulation of polymorphonuclear neutrophils (PMN) in the lungs at the early stage of burns. METHODS: Myeloperoxidase content in lung tissues and bronchoalveolar lavage fluid (BALF) were detected. ICAM-1 and its mRNA expression in lung tissues were determined by immunohistochemical method and in situ hybridization. CD11b/CD18 expression on the peripheral PMNs was measured by flow cytometry. RESULTS: The levels of myeloperoxidase in lung tissues and BALF after burn injury were markedly higher than those of control. Expression of ICAM-1 and its mRNA in the lung tissues and CD11b/CD18 on peripheral PMNs surface was significantly increased at 2, 6, 12, 24 h after burns. CONCLUSIONS: PMNs accumulation in the lungs is related to increased ICAM-1 expression on pulmonary microvascular endothelial cells and CD11b/CD18 expression on PMN at the early stage of burn injury. PMID- 12749788 TI - Early period of fracture healing in ovariectomized rats. AB - OBJECTIVE: To evaluate the effect of osteoporosis on fracture healing through observing the histomorphological changes, bone mineral density of callus and expression and distribution of transforming growth factor beta 1 (TGF-beta1), basic fibroblast growth factor (bFGF) and bone morphogenetic protein-2 (BMP-2) in ovariectomized rats. METHODS: Sixty female Sprague-Dawley rats (aged 12 weeks and weighing 235 g on average) were randomly divided into an ovariectomized (OVX) group (n=30) and a sham-operated (SO) group (n=30). Ovariectomy was performed in the OVX rats and same incision was made in the SO rats. Three months later, fracture of femoral shaft was made on all the rats. Then they were killed at different time points. Callus formation was observed with histological and immunohistochemical methods. RESULTS: A reduction in callus and bone mineral density in the healing femur and a decrease of osteoblasts expressing TGF-beta1 near the bone trabecula were observed in the OVX rats 3-4 weeks after fracture. Histomorphological analysis revealed a higher content of soft callus in the OVX rats than that in the SO rats. Immunohistochemistry results showed that no remarkable difference in expression and distribution of BMP-2 and bFGF between the OVX and SO groups was found. CONCLUSIONS: Osteoporosis influences the quantity and quality of callus during the early period of fracture healing. The effect of osteoporosis on fracture healing has no relationship with the expression of BMP-2 or bFGF. The decreased expression of TGF-beta1 in osteoblasts may cause a decrease in quality of fracture healing after osteoporosis. PMID- 12749789 TI - Traumatic endophthalmitis following penetrating ocular injuries with retained intraocular foreign bodies. AB - OBJECTIVE: To evaluate the outcome and analyze the methods of surgical treatment of traumatic endophthalmitis following penetrating eye injuries with retained eye foreign bodies. METHODS: A total of 62 consecutive cases (58 men, 4 women) from January 1999 to December 2001 with IOFBs following penetrating eye injuries were retrospectively studied. The ages ranged from 8 to 46 years (mean 23 years). Sixty patients (63 eyes) underwent pars plana vitreotomy and 1 patient underwent external magnet extraction. The follow-up ranged from 3 to 36 months (mean 12.5 months). RESULTS: Ten eyes developed endophthalmitis, among which 7 (10.94%) were diagnosed preoperatively. The most frequently cultured organism was Staphylococcus epidermis (44.44%, 4/9). Postoperatively, retinal detachment due to vitreoretinal proliferation occurred in 5 patients with endophthalmitis and in 9 patients without endophthalmitis. All the retinal detachments were reattached with additional vitreoretinal surgery. Two eyes with endophthalmitis and two without endophthalmitis were eviscerated. CONCLUSIONS: Post-traumatic endophthalmitis with intraocular foreign bodies (IOFBs) deserves great attention because of its high incidence and poor prognosis. Vitrectomy is suggested for the treatment of IOFBs and its complications, and it should be performed as soon as possible. Routine intravenous administration of antibiotics combined with periocular injection and topical antibiotics postoperatively are recommended. PMID- 12749790 TI - Thoracic spinal trauma associated with closed thoracoabdominal injury. AB - OBJECTIVE: To explore the management of thoracic spinal trauma (TST) associated with closed thoracoabdominal injuries (CTAI). METHODS: A retrospective study was made on 259 patients with TST admitted to our department as an emergency treatment from January 1996 to June 2001. We summarized the clinical features of TST associated with CTAI. RESULTS: Among 259 patients with thoracic spinal trauma, 112 were associated with CTAI. Traffic accident was the most common cause. The force causing upper TST was more violent than that causing the lower. Pulmonary complications were the leading cause of death in this group. Surgery could not improve neurological function for completely paraplegic patients. CONCLUSIONS: The reason that upper TST has the tendency to be associated with CTAI is its special anatomical feature. Routine ultrasonic examination can avoid misdiagnosis of latent closed abdominal injuries associated with spinal injury. The presence of potential injuries, especially CTAI, should be considered when deciding whether or not to perform surgery early. PMID- 12749791 TI - Novel multi-probe RNase protection assay set for detection of endotoxin associated receptors gene expression. AB - OBJECTIVE: To construct the multi-probe ribonuclease protection assay (RPA) template set to be used for detecting expression patterns of MD-2, TLR4, CD14 mRNAs in human peripheral blood mononuclear cells. METHODS: The designed cDNA fragments of the three genes were generated by polymerase chain reaction (PCR) using specific primers and directionally cloned into EcoR I and Hind III sites of expression plasmid pSP72 containing the T7 promoter, the linearized plasmids was used as template to synthesize anti-sense RNA probes. Then we extracted total RNA from peripheral blood mononuclear cells (PBMC) and detected the dynamic expression patterns of the three genes with RPA method. RESULTS: The proper sequence and orientation of the template set were confirmed by sequencing and the template set was successfully used to assay TLR4, MD-2 and CD14 mRNAs in human PBMC. The results showed that the three detected genes decreased transiently 1-3 hours after 100 ng/ml LPS stimulation. CONCLUSIONS: These new RPA multi-probe set provided valuable tool for the simultaneous quantitative determination of expression of TLR4, CD14 and MD-2 mRNAs in both constitutive and inducible types. PMID- 12749792 TI - Clinical analysis of the syndrome of inappropriate antidiuretic hormone secretion after brain injury. AB - OBJECTIVE: To study the diagnosis and treatment of the syndrome of inappropriate antidiuretic hormone secretion (SIADH) after brain injury. METHODS: A retrospective analysis was conducted on 12 patients suffered from SIADH after brain injury. The clinical features of these patients were similar to those of common hyponatremia. Most of the hyponatremia were detected by routine examinations. Supplement of salt as the initial treatment was used in these patients. If natremia did not rise or descended 2-3 days after treatment, SIADH was considered or diagnosed. Treatment scheme should be adjusted to limit water and natrium instead of supplying salt. Frusemide and albumin were the first choice for dehydration therapy. RESULTS: 24-48 hours after limiting water and natrium, 12 patient's natremia level was back to normal. Eight out of 12 patients were corrected completely in 1 week, 1 in 14 days, and 1 in 3 months after injury. CONCLUSIONS: Diagnosis of SIADH is difficult before treatment though effective treatment can be obtained if we adopt correct strategy. In these patients, the diagnosis of SIADH is confirmed in the course of treatment. PMID- 12749793 TI - Treatment of open supracondylar fracture of humerus. AB - OBJECTIVE: To explore the injury mechanism and treatment principle of open supracondylar fracture of humerus. METHODS: The data of 32 patients with open supracondylar fracture of humerus hospitalized in our department in the recent 20 years were analyzed retrospectively. On an average, they were followed up for 30 months. The relationship between the fracture type, situation of wounds, operating time, operating method and time for postoperative functional exercise and final function of elbow joint were evaluated with Flynn scoring standard. RESULTS: Excellent therapeutic effect was found in 14 cases, good in 12 and bad in 6, with the percentage of fine therapeutic effect of 81.25%. CONCLUSIONS: For the patients with open supracondylar fracture of humerus, debridement and internal fixation should be made as early as possible, for thorough debridement and correct internal fixation are the key points to improve the prognosis. PMID- 12749794 TI - Traumatic interhemispheric subdural hematoma. PMID- 12749795 TI - Treatment of 116 patients with severe craniocerebral injury in basic-level hospital. PMID- 12749796 TI - Of oysters and man. PMID- 12749797 TI - Vertically transmitted herpesvirus infections. AB - The importance of recognizing perinatally acquired herpes simplex virus infections is now well understood and recently, increased attention has been devoted by clinicians to the problem of perinatally acquired cytomegalovirus infections. Less commonly, other members of the herpesvirus family may also be transmitted from mother to infant in the perinatal period, causing disease. Future developments of the management of herpesvirus infections will include greater roles for antiviral therapies and vaccines. Expanded use of oral nucleoside antiviral therapies in pregnancy has engendered uncertainty in how best to screen, monitor, and treat women at risk of transmitting the infection. Vaccines against herpesviruses have progressed in clinical trials, and the imminent licensing of such vaccines will further complicate clinical management decisions. As potential therapeutic agents become available, better understanding and recognition of risk factors, and awareness of the clinical presentation of perinatally acquired herpesvirus infections, will be required to maximize satisfactory pregnancy outcomes. PMID- 12749798 TI - Immunosuppressive effects of beta-herpesviruses. AB - Immunomodulatory effects of human beta-herpesviruses have been reported in vitro and in vivo. Clinical studies suggest that beta-herpesvirus infection may increase the risk for other infections, the severity of infection, or the tempo of disease progression. An increased incidence of bacterial and fungal infections, and graft rejection, have been reported in association with cytomegalovirus (CMV), and human herpesviruses type 6 and type 7 infections have been implicated as risk factors for CMV infection and graft rejection. Beta herpesviruses may also interact with HIV-1 and hepatitis C. To prove a causal relationship between beta-herpesviruses and specific clinical outcomes, randomized trials, with selective suppression of the virus, are required. Such trials have been performed for CMV and showed a reduction in bacterial and fungal infections as well as rejection in selected solid organ transplant recipients. More trials are needed to evaluate whether the effects seen in observational studies are truly related. PMID- 12749800 TI - [What can pleural fluid tell us?]. PMID- 12749801 TI - [Drug treatment of chronic obstructive pulmonary disease on two levels of patient care: degree of compliance with recommended protocols]. AB - OBJECTIVE: The aims of this study were to identify the drug treatment protocols applied by primary care physicians or pneumologists for patients with stable chronic obstructive pulmonary disease (COPD) in Spain, to determine the agreement between prescription practices and current recommendations and to assess differences between the two levels of patient care. PATIENTS AND METHODS: The study was observational, descriptive and multicenter. A stratified random sample of patients treated by family physicians or pneumologists was taken for every region in Spain. RESULTS: Five hundred sixty-eight (63.2%) of the 898 subjects fulfilled COPD diagnostic criteria; 100 were treated by primary care physicians and 460 by pneumologists. In 8 cases the caregiver was unknown. Obstruction was mild-to-moderate in 144 cases and severe in 416. The drugs most commonly prescribed were ipratropium bromide (77.8%), inhaled short-acting beta(2) agonists (65.8%), inhaled corticosteroids (61.0%), long-acting beta(2) agonists (46.4%) and theophyllines (41.3%). Primary care physicians prescribed inhaled short-acting beta 2-agonists most often, whereas pneumologists prescribed anticholinergics most often. In the primary care setting, no differences in treatment protocols were observed based on severity of COPD, degree of dyspnea or quality of life. More consistent differences were seen in treatment by pneumologists. In both settings, prescription was more frequently given when COPD was severe. The most commonly prescribed inhalation device was the Turbuhaler in primary care and the pressurized canister in pneumology. CONCLUSIONS: Treatments prescribed for COPD patients do not follow current guidelines strictly, particularly in the primary care setting. Different prescription protocols are used at the different levels of patient care. PMID- 12749802 TI - [Reliability of chronic obstructive pulmonary disease diagnosis by primary care physicians and pneumologists in Spain. Predictive factors]. AB - OBJECTIVE: The aims of this study were to assess the methods used by primary care physicians and pneumologists to diagnose chronic obstructive pulmonary disease (COPD) in Spain, and to analyze the factors affecting correct diagnosis of the disease. MATERIAL AND METHODS: This observational, descriptive, cross-sectional and multicenter study enrolled a stratified randomized sample from each Spanish region from the practices of primary care physicians and pneumologists. RESULTS: Five hundred sixty-eight (63.2%) of the 898 subjects enrolled had airway obstruction, 92 (10.3%) did not fulfill functional criteria for COPD and 238 (26.5%) did not perform spirometric tests to confirm the diagnosis and establish severity of disease. Primary care physicians classified 29.3% of the patients correctly, whereas pneumologists diagnosed 84.8% correctly. Clinical and/or radiologic criteria were the basis for correct diagnosis in 38.6% of the cases managed by primary care physicians and 10.2% of those treated by pneumologists. Spirometry was available to 49.1% of the primary care physicians and 97.8% of the pneumologists' cases (p < 0.001). Moreover, only 29.9% of the primary care settings had a technician in charge of performing the study, in comparison with 97.8% of the specialized pneumology settings (p < 0.001). The use of spirometry in diagnosing COPD was related to level of patient care (primary or specialized), availability of the test in the primary care setting, place of residence and a patient's situation of temporary absence from work due to COPD. CONCLUSIONS: Many COPD patients are incorrectly diagnosed, particularly in primary care. There are differences in diagnostic procedures at the different levels of patient care. The availability of spirometry is an important factor for correctly diagnosing COPD. PMID- 12749803 TI - [Lung cancer in the health care area of A Coruna (Spain): incidence, clinical approach and survival]. AB - OBJECTIVE: To determine the incidence, clinical approach, diagnostic delay and survival for bronchogenic carcinoma (BC) in the public health area of A Coruna (Spain). PATIENTS AND METHOD: This was a retrospective study of patients with a diagnosis of BC made in 1995 and 1996 in a health care area with 509,000 inhabitants. For analysis we gathered demographic, clinical and cyto-histologic data and analyzed diagnostic delay, extension of disease and time of death. Crude, age-specific and age-standardized incidences were calculated. RESULTS: BC was diagnosed in 378 patients (95% men and 5% women). The crude incidence was 37 per 100,000 inhabitants (73.7 per 100,000 men and 3 per 100,000 women) and the age-standardized incidence was 21.7 per 100,000. Diagnosis was by cyto-histology in 87% of the cases, and the most frequent biopsy finding was epidermoid carcinoma. Diagnosis was by clinical or radiologic findings for 13.7%, and patients in that group were older and had greater comorbidity. No symptoms were present in 13% and in those patients diagnosis was earlier, with non-small BC predominating and 60% treated surgically. The diagnostic delay (time from the first symptom to histologic confirmation) was 2.5 months (median, 2.1), and length of delay did not affect survival. Surgery was performed in 23% of the patients with non-small BC. The median survival was 7.1 months (5.2 months for patients who were not treated surgically, and 37.6 for those who were). CONCLUSIONS: The incidence of BC in men in our study is consistent with that reported for other Spanish regions, although the incidence we observed for women is much lower. Diagnostic delay did not affect survival. The percentage of patients with surgical-stage disease is very low and survival is very short. These findings support the need for early diagnosis. PMID- 12749804 TI - [Lobectomy versus limited resection to treat non-small cell lung cancer in stage I: a study of 78 cases]. AB - OBJECTIVE: To compare mortality and loco-regional recurrence rates in patients treated surgically for stage I non-small cell lung cancer by either lobectomy or sublobar lung resection (pulmonary wedge) from 1994 to 1998. PATIENTS AND METHODS: Seventy-eight patients who underwent lung resection for stage I non small cell lung cancer from January 1994 to December 1998 were reviewed retrospectively. Fifty patients underwent lobectomy and 28 limited resection (pulmonary wedge). We reviewed oncologic history, tumor histology, loco-regional recurrence and tumor-related mortality during the follow-up period. RESULTS: Histology, lobectomies: 36 (72%) squamous cell carcinomas, 12 (24%) adenocarcinomas and 2 (4%) large cell carcinomas. Histology, wedge resection: 12 (42.8%) squamous cell carcinomas and 16 (57.2%) adenocarcinomas. Staging, lobectomies: 15 Ia (30%) and 35 Ib (70%). Staging, wedge resections: 16 Ia (57.2%) and 12 Ib (42.8%). Loco-regional recurrence: lobectomies 9 (18%) and wedge resections 4 (14.3%) plus 1 remote metastasis (3.6%). Mortality during follow-up: lobectomies 8 (16%) and wedge resections 4 (14.3%). Accumulated survival rate (in months): lobectomies 62.38 and wedge resections 63.92. The rates of loco-regional recurrence and accumulated survival in the two groups were not significantly different. CONCLUSIONS: The rates of loco-regional recurrence and accumulated survival in this study agree with those from other retrospective studies. We found no statistically significant differences between patients undergoing lobectomy and those undergoing sublobar resection. Wedge resection is an appropriate technique for patients with stage I non-small cell lung cancer who can not tolerate lobectomy. PMID- 12749805 TI - [Asthmatic characteristics in smokers with advanced emphysema]. AB - OBJECTIVES: Most pneumologists assume that chronic asthma and emphysema associated with smoking differ both in their etiology and type of inflammation. However, it is difficult to study subjects who have both diseases simultaneously. We designed a prospective study to determine whether or not some advanced emphysema patients who are smokers have an asthmatic profile suggesting the two diseases overlap. PATIENTS AND METHOD: Twenty-three smokers with moderate to severe obstruction and radiologic signs of advanced emphysema were studied. The following signs of asthma were analyzed for all patients: positive acute response to bronchodilators, eosinophilia in peripheral blood, significant eosinophilia in sputum, significant total serum immunoglobulin E (IgE) and specific IgE positive for common allergens and fungi. An asthmatic profile was defined as the presence of 3 or more of the aforementioned signs. Statistical analysis was by calculation of the confidence interval on the total number of positive cases to estimate the population percentage. RESULTS: Data on at least 4 characteristics were available for 21 of the 23 patients. Ten (48%) of the 21 had asthmatic profiles (3 or more of the aforementioned characteristics). This finding suggests that asthmatic characteristics are present in between 17.6 and 79.6% of the studied population with advanced emphysema, with a confidence interval of 99.9%. CONCLUSIONS: We conclude that emphysema initially associated with smoking may mean that at least 17.6% of patients have a clear asthmatic profile, which indicates a high degree of overlap between asthma and emphysema in smokers. Thus, our findings are consistent with the Dutch theory that chronic obstructive diseases have a common origin in underlying allergic sensitization and bronchial hyperresponsiveness alongside various other influential factors such as tobacco smoking over the course of disease. PMID- 12749806 TI - [Series 4: respiratory muscles in neuromuscular diseases and the chest cavity. Decision making in the clinical management of patients with lateral amyotrophic sclerosis]. PMID- 12749808 TI - [Pasteurella multocida infection of cavitated lung squamous carcinoma]. AB - Pasteurella multocida has rarely been reported to cause lung disease in humans. Infection usually arises from bites or scratches from animal carriers of the pathogen. Cases of pneumonia, lung abscess, airway infection or infection of pre existing bronchiectasis have been described, usually in individuals who are in direct contact with carrier animals and who have a chronic debilitating disease. It is unusual for P. multocida to be ingested and appear among oropharyngeal flora in humans. We report the first case published (Medline search 1966-2002) of a cavitated lung with squamous carcinoma that became infected by P. multocida in an elderly patient who denied contact with potential carrier animals. We believe that the P. multocida infection in humans is underdiagnosed because clinical suspicion is low and the bacterium is highly susceptible to common antibiotics. PMID- 12749807 TI - [Bronchial-associated lymphoid tissue lymphoma]. AB - Two cases of non-Hodgkin's endobronchial or bronchial-associated lymphoid tissue lymphoma are reported; such cases are either extremely rare or underestimated. We emphasize the need to perform endoscopic examination in patients with lymphoma and clinical findings that suggest bronchial disease. PMID- 12749809 TI - [Burkholderia cepacia pneumonia in a patient with common variable immunodeficiency]. PMID- 12749810 TI - [Minimally invasive correction of pectus excavatum by video-assisted thoracoscopy]. PMID- 12749811 TI - [Lung toxicity due to thalidomide]. PMID- 12749812 TI - [Hospitalization of patients with community-acquired pneumonia]. PMID- 12749813 TI - Trichothecenes in the environment: relevance to human health. AB - Trichothecenes are agriculturally important mycotoxins of relevance to human health. Fungi capable of producing trichothecenes can be found throughout the world, and include certain species of Fusarium, Myrothecium, and Stachybotrys. The production of mycotoxins by these toxigenic species is determined by genetic factors and the environmental conditions of their growth. The environmental fate of trichothecenes may be affected by other microorganisms that can detoxify them. Deoxynivalenol and T-2 toxin are examples of trichothecenes that are detectable as natural and unavoidable contaminants of certain agricultural commodities as well as commercial foods. Current estimates of dietary exposure to deoxynivalenol and T-2 toxin are below thresholds for adverse effects that have been reported in experimental animal studies, although historical epidemics of human illness have rarely been described in association with consumption of food derived from heavily contaminated grains. The toxicodynamic properties of trichothecenes include inhibition of protein synthesis and immunomodulatory effects. Very little information is available relating to their toxicokinetics and toxicodynamics in humans. While there is general agreement that the diet represents an important source of human exposure to trichothecenes, risk assessment from non-dietary routes of exposure is complicated by the limited epidemiological data that are currently available. PMID- 12749814 TI - Upper airway irritation of terpene/ozone oxidation products (TOPS). Dependence on reaction time, relative humidity and initial ozone concentration. AB - Recently, we reported the formation of unidentified strong upper airway irritants in reaction mixtures of terpenes and ozone. The identified products included aldehydes, ketones and carboxylic acids. Here we report the effects of variation of reaction time, relative humidity and initial ozone concentration on irritant formation in a flow reaction system using R-(+)-limonene and isoprene. Upper airway irritation was measured in mice as reduction of the respiratory rate. For both substances maximum irritation was observed for low humidity (<2% RH)/short time (16-30 s) reaction mixtures, and both moderate humidity ( approximately 32% RH) and longer reaction times (60-90 s) resulted in significantly less irritation. These results suggest that some unidentified intermediates react with water vapor to give less irritating products. Irritation measured at four ozone concentrations (0.5, 1, 2 and 3.5 ppm) using low humidity/short time reaction conditions for limonene (50 ppm) and isoprene (500 ppm) revealed that at 0.5 ppm, irritation was at the same level as that for the pure terpenes, indicating that at 0.5 ppm ozone the combined irritant effect was near the no effect level for the product mixture. PMID- 12749815 TI - Effect of alpha-ketoglutarate and oxaloacetate on brain mitochondrial DNA damage and seizures induced by kainic acid in mice. AB - The effects of alpha-ketoglutarate and oxaloacetate on brain mitochondrial DNA (mtDNA) damage and seizures induced by kainic acid were examined both in vivo and in vitro. An intraperitoneal (ip) injection of kainic acid (45 mg/kg) produced broad-spectrum limbic and severe sustained seizures in all of the treated mice. The seizures were abolished when alpha-ketoglutarate (2 g/kg) or oxaloacetate (1 g/kg) was injected intraperitoneally in the animals 1 min before kainic acid administration. In addition, the administration of kainic acid caused damage to mtDNA in brain frontal and middle cortex of mice. These effects were completely abolished by the ip preinjection of alpha-ketoglutarate (2 g/kg) or oxaloacetate (1 g/kg). In vitro exposure of kainic acid (0.25, 0.5 or 1.0 mM) to brain homogenate inflicted damage to mtDNA in a concentration-dependent manner. The damage of mtDNA induced by 1.0 mM kainic acid was attenuated by the co-treatment with alpha-ketoglutarate (2.5 or 5.0 mM) or oxaloacetate (0.75 or 1.0 mM). Furthermore, in vivo and in vitro exposure of kainic acid elicited an increase in lipid peroxidation. However, the increased lipid peroxidation was completely inhibited by cotreatment of alpha-ketoglutarate or oxaloacetate. These results suggest that alpha-keto acids such as alpha-ketoglutarate and oxaloacetate play a role in the inhibition of seizures and subsequent mtDNA damage induced by the excitotoxic/neurotoxic agent, kainic acid. PMID- 12749816 TI - DNA repair gene XPD and susceptibility to arsenic-induced hyperkeratosis. AB - Chronic exposure to inorganic arsenic is known to cause non-melanocytic skin and internal cancers in humans. An estimated 50-70 million people in Bangladesh have been chronically exposed to arsenic from drinking water and are at risk of skin and other cancers. We undertook the first study to examine whether genetic susceptibility, as determined by the codon 751 SNP (A-->C) of the DNA repair gene XPD, influences the risk of arsenic-induced hyperkeratotic skin lesions, precursors of skin cancer, in a case-control study of 29 hyperkeratosis cases and 105 healthy controls from the same community in an area of Bangladesh. As expected, there was a monotonic increase in risk of hyperkeratosis in relation to urinary arsenic measures but the XPD genotype was not independently associated with the risk. However, the increase in hyperkeratosis risk in relation to urinary arsenic measures genotype was borderline significant for urinary total arsenic (P for trend=0.06) and statistically significant for urinary creatinine adjusted arsenic (P for trend=0.01) among subjects with the XPD A allele (AA) but not among subjects with the other XPD genotypes. Among AA carriers, the risk for the highest arsenic exposed group compared with the lowest was more than 7-fold for urinary total arsenic and about 11-fold for urinary creatinine adjusted arsenic. In conclusion, our findings suggest that the DNA repair gene XPD may influence the risk of arsenic-induced premalignant hyperkeratotic skin lesions. Future larger studies are needed to confirm this novel finding and investigate how combinations of different candidate genes and/or other host and environmental factors may influence the risk of arsenic induced skin and other cancers. PMID- 12749818 TI - Oxidative DNA damage following exposure to dimethylarsinous iodide: the formation of cis-thymine glycol. AB - The purpose of the present study was to elucidate the genotoxic mechanism of trivalent dimethylated arsenic, particularly the induction mechanism of oxidative stress in nuclear bases. Cis-thymine glycol was used as a biomarker of DNA oxidation damage. The treatment of thymine with dimethylarsinous iodide (DMI), a model compound of dimethylarsinous acid, induced the formation of cis-thymine glycol. This oxidative damage was induced via the production of dimethylated arsenic peroxide, but not via the production of superoxide anion or hydrogen peroxide. Trivalent dimethylated arsenic may thus play an important role in arsenic carcinogenesis through the induction of oxidative base damage. PMID- 12749817 TI - Thymoquinone protects against experimental colitis in rats. AB - The present work was done to investigate the possible effects of thymoquinone on acetic acid-induced colitis in rats. Colitis was induced by intracolonic injection of 3% acetic acid. Several parameters including macroscopic score, histopathological and biochemical, were determined to assess the degree of protection. Biochemical parameters such as myeloperoxidase activity, reduced glutathione levels, platelet activating factor (PAF) and histamine were measured following standard assay procedures. The study showed that pretreatment of rats for 3 days with thymoquinone (10 mg/kg) was able to give complete protection against acetic acid-induced colitis an effect significantly higher than sulfasalazine (500 mg/kg) control group. The smaller dose of thymoquinone (5 mg/kg) produced partial protection. Moreover, the biochemical and histopathological changes were reversed and brought towards the control. These results suggest a beneficial effect of thymoquinone against experimentally induced colitis and the possible mechanism of the protective effects may be partly due to an antioxidant action. PMID- 12749819 TI - Patterns of gene expressions induced by arsenic trioxide in cultured human fibroblasts. AB - Arsenic exposure is associated with several human diseases and particularly, with neoplasia. Although the mechanism of arsenic toxicity is not fully understood, several recent works pointed out the involvement of oxidative stress in arsenic induced DNA damage that, in living cells, correlates with changes in gene expressions. In cultured human fibroblasts exposed for 24 h to micromolar arsenic concentrations, we studied, using real-time RT-PCR, the expression profile of a limited number of genes: genes coding for a stress protein (HSP70), transcription factors (cJUN, cFOS, ETR103, ETR101 and TTP) and cell cycle or DNA repair proteins (P21, GADD153). We observed that the expression profile of genes followed individual different patterns that can be summed up in early-transient gene expression by contrast to delayed gene expression. PMID- 12749820 TI - The effects of mono-2-ethylhexyl phathalate, adriamycin and N-ethyl-N-nitrosourea on stage-specific apoptosis and DNA synthesis in the mouse spermatogenesis. AB - Tubule segments from defined stages of the mouse seminiferous epithelium were incubated for 4, 8 and 24 h in the presence of different concentrations of mono-2 ethylhexyl phthalate (MEHP), adriamycin, and N-ethyl-N-nitrosourea (ENU) to study the effects of these chemicals on germ cell apoptosis and DNA synthesis. Apoptosis was measured by using the in situ end-labeling (ISEL) technique and microscopic analysis of apoptotic cells, and DNA synthesis was assessed by 3H thymidine incorporation. In the mouse testis after 8 h incubation, MEHP increased the number of apoptotic cells significantly at 0.01 and 0.1 mM concentrations in stages IX-XI of the mouse seminiferous cycle. Adriamycin increased the number of apoptotic cells significantly at 0.1 and 1.0 microM concentrations in stages IX XI. In tubules from stages IX-XI cultured in the presence on 0.01 and 0.1 mM ENU the number of apoptotic cells was increased significantly. In dose-response experiments trend of decreasing amount of DNA synthesis was observed, but no statistical differences were found. MEHP, adriamycin and ENU increased the amount of apoptotic cells in model where transillumination assisted microdissection was combined to ISEL. We conclude that ISEL technique provides a useful method to study the stage-specific effects of chemicals on spermatogenesis. PMID- 12749821 TI - Nicotine accumulation in the mouse brain is age-dependent and is quantitatively different in various segments. AB - Mice, varying from newborn to adult mice, were injected intraperitoneally with 1 mg/kg of 14C-nicotine. The nicotine concentration in blood and different segments of the brain (Cerebellum, Pons and Medulla oblongata, Midbrain, B. olfactory, Hypothalamus, Hippocampus and Cortex) was determined in newborn (0-day old); in 1 , 3-, 5-, 7-, 9-, 12- and 17-day old; and in adult (26-day old) mice. The nicotine concentration in the blood did not differ in mice aged 0-12 days, but the concentration thereafter decreased substantially until adult age. The decrease was associated with metabolism of nicotine as evidenced by cotinine formation, as well accumulation in different regions of the brain. Irrespective of the small continuous brain growth from day 1 until adult age, the age dependent nicotine uptake was dramatically increased in all segments of the brain between 9 and 12 days of age. However, the various brain regions showed differences in magnitude of growth and age-dependent changes in the distribution of nicotine. The relative nicotine concentration in Pons Med. obl. seems to be high in young mice until 9 days of age; thereafter, the concentration decreases in comparison to the other regions. Conversely, the relative concentration in the hippocampus and cortex rises from day 9 to 17 of age. The present results show that the ability of the brain to accumulate nicotine increases with age, reaching a peek at day 12 of age. Furthermore, this study for the first time shows that the age-dependent brain nicotine accumulation is quantitatively different in the various segments of the brain. PMID- 12749822 TI - Involvement of MEK/ERK pathway in cephaloridine-induced injury in rat renal cortical slices. AB - We have previously reported that free radical-mediated injury induced by cephaloridine (CER) is enhanced by phorbol 12-myristate 13-acetate (PMA), a protein kinase C (PKC) activator, in rat renal cortical slices. We have also shown that PKC activation in mitochondria is involved in CER-induced nephrotoxicity in rats. We investigated the role of a downstream PKC pathway, a MEK/ERK pathway, in free radical-induced injury in rat renal cortical slices exposed to CER. Immediately after preparing slices from rat renal cortex, the slices were incubated in the medium containing MEK inhibitors. ERK1/2 activation was determined by Western blot analysis for phosphorylated ERK (pERK) 1/2 protein in nucleus fraction prepared from the slices exposed to CER. Prominently, CER caused not only increases in lipid peroxidation as an index of free radical generation and in LDH leakage as that of cell injury in the slices, but also marked activation of ERK1/2 in nucleus fraction. PD98059 and U0126, MEK1/2 inhibitors, significantly attenuated CER-induced increases in lipid peroxidation and LDH leakage in the slices. PD98059 also suppressed ERK1/2 activation in nucleus fraction prepared from the slices treated with CER. Inhibition of other MAP kinase pathways, p38 MAP kinase and c-Jun N-terminal kinase (JNK) had no effect on CER-induced increases in lipid peroxidation level and LDH leakage in the slices. The present results suggest that a MEK/ERK pathway down stream of a PKC pathway is probably involved in free radical-induced injury in rat renal cortical slices exposed to CER. PMID- 12749823 TI - Screening of medicinal plants used in South African traditional medicine for genotoxic effects. AB - Dichloromethane and 90% methanol extracts from 51 South African medicinal plants were evaluated for potential genotoxic effects using the bacterial Ames and VITOTOX tests with and without metabolic activation. Dichloromethane extracts from bulbs of Crinum macowanii showed mutagenicity in strain TA98 with and without metabolic activation, whereas extracts from leaves of Chaetacme aristata and foliage of Plumbago auriculata showed mutagenicity and/or toxicity. Extracts from the leaves of Catharanthus roseus and twigs of Combretum mkhzense were mutagenic with metabolic activation only. The only 90% methanol extracts that were mutagenic in strain TA98 were from the leaves of C. roseus and Ziziphus mucronata in the presence of metabolic activation. No genotoxic effects were found in strain TA100 or in the VITOTOX test. PMID- 12749824 TI - The effects of tannic acid on cytochrome P450 and phase II enzymes in mouse liver and kidney. AB - Tannic acid, a naturally occurring plant polyphenol, was shown to decrease the mutagenicity and/or carcinogenicity of several amines derivatives and polycyclic aromatic hydrocarbons in rodents. The aim of this study was to evaluate the effect of tannic acid on the activities of murine cytochrome P450 and phase II enzymes. The activities of ethoxyresorufin-O-deethylase (EROD), methoxyresorufin O-demethylase (MROD), p-nitrophenol hydroxylase (PNPH), glutathione S-transferase (GST), UDP-glucuronosyltransferase (UDPGT) and NAD(P)H:quinone oxidoreductase (NQO1) were measured in the liver and kidney microsomes of female Swiss mice treated intraperitoneally (i.p.) with tannic acid in the dose range of 20-80 mg/kg. At the highest dose, tannic acid decreased the activities of EROD and MROD by 25-28% in mouse liver, while the activity of both hepatic and renal PNPH was reduced by approximately 50% as result of treatment. Moreover, Western blot analysis with CYP2E1 specific antibody showed a significant decrease in the levels of hepatic CYP2E1 in tannic acid treated animals. This polyphenol affected also the phase II enzymes in both tissues examined. The activity of GST was elevated in kidneys, but reduced in livers of the animals treated with tannic acid. The most striking effect was the inhibition of hepatic NQO1. The effect was dose dependent and almost 90% inhibition was observed after the treatment with tannic acid at the dose of 60 and 80 mg/kg. The same treatment caused the approximately 60% inhibition of renal NQO1. These results indicate that tannic acid, beside of scavenging active metabolites of chemical carcinogens, can change their metabolism by modulating the enzymes involved in xenobiotics activation and/or detoxification pathways. PMID- 12749825 TI - Effects of flutamide in the rat testis on the expression of occludin, an integral member of the tight junctions. AB - In an effort to uncover the gonadal impairment by the antiandrogen flutamide (FM) in males, the effect of subacute administration of FM on the expression of tight junction (TJ) genes that build the blood-testis barrier (BTB) was investigated in adult rat testis. At 13 weeks old of age male rats were given vehicle (corn oil) or FM (25 mg/kg per day, in corn oil) orally for 6 days. At 8 days (D8) after the first dose, testicular expression of the occludin, claudin-1, and -11 was analyzed by semiquantitative RT-PCR. The testicular weight of the FM-treated rats on D8 was a little but significantly higher than in the control group. On D8 the expression of occludin in the FM-treated animals was significantly decreased but claudin-1 and -11 were not altered significantly. Because FM administration inhibits germ cell differentiation, it is likely that the down-regulated occludin expression in FM-rat testes may be attributed to the alteration in the paracrine interaction between Sertoli cells and germ cells in testis. It also emphasized that FM might have differentially affected the transcription of TJ genes in Sertoli cells building the BTB. These findings provide a rationale for a number of observations on the gonadal impairment by FM in males and suggest that FM is potentially harmful to spermatogenesis by alteration of the BTB. PMID- 12749827 TI - Protective effects of progesterone on implantation failure induced by dibutyltin dichloride in rats. AB - We previously showed that dibutyltin dichloride (DBTCl) at 7.6 mg/kg and higher on days 0-3 of pregnancy caused implantation failure and a decline in serum progesterone levels in rats and hypothesized that the decline is responsible for the implantation failure. This study was conducted to determine the protective effects of progesterone on the DBTCl-induced implantation failure in rats. Rats were given oral DBTCl at 0, 7.6, or 15.2 mg/kg on days 0-3 of pregnancy and/or subcutaneous progesterone at 2 mg/rat on days 0-8 of pregnancy. The reproductive outcome was determined on day 9 of pregnancy. No effects of administration of progesterone alone on the pregnancy rate and number of implantations were found. The pregnancy rate and number of implantations were significantly decreased after administration of DBTCl alone. The pregnancy rate and number of implantations were higher in the groups given DBTCl and progesterone than the groups given DBTCl alone. The present data indicate that progesterone protects, at least in part, against the DBTCl-induced implantation failure and support our hypothesis that the decline in progesterone levels is a primary mechanism for the implantation failure due to DBTCl. PMID- 12749826 TI - Benzo(a)pyrene-coated onto Fe2O3 particles-induced apoptotic events in the lungs of Sprague-Dawley rats. AB - Because benzo(a)pyrene (B(a)P)-coated onto hematite (Fe(2)O(3)) particle-induced adverse effects might alter cell homeostasis in lungs, we investigated the induction of some apoptotic events by such a concurrent exposure on this relevant organ target. Sprague-Dawley rats were intratracheally instilled with Fe(2)O(3) (3 mg), B(a)P (3 mg) or B(a)P (3 mg)-coated onto Fe(2)O(3) particles (3 mg). Forty-eight hours later, both the tumor necrosis factor-receptor and the mitochondrial pathways were studied. We found that exposure to B(a)P (1.13-fold, P<0.05) or to B(a)P-coated onto Fe(2)O(3) particles (1.15-fold, P<0.05) increased caspase 3 activity. However, only the concurrent exposure activated both the caspases 8 (1.21-fold, P<0.05) and 9 (1.27-fold, P<0.05). After exposure to either chemical alone, there was a discrepancy between the findings on tumor necrosis factor-alpha and caspase 8, on one hand, and on cytochrome c and caspase 9, on the other hand. Hence, we suggested that the oxidative stress induced by Fe(2)O(3) or B(a)P will continuously lower or deplete caspase activities, thereby reducing or even avoiding the activation of the apoptotic pathways. In addition, transcriptional induction of p53 gene by Fe(2)O(3) (1.73-fold, P<0.01) or B(a)P coated onto Fe(2)O(3) particles (1.53-fold, P<0.01) was observed. Taken together, the present results support the underlying hypothesis that the influence of Fe(2)O(3) in B(a)P/Fe(2)O(3) mixtures on the ability of B(a)P to induce some of the events firmly involved in the apoptotic pathways will also be one of the ways that Fe(2)O(3) can affect B(a)P toxicity in lungs. PMID- 12749828 TI - Calcitonin gene-related peptide as inflammatory mediator. AB - Sensory neuropeptides have been proposed to play a key role in the pathogenesis of a number of respiratory diseases such as asthma, chronic obstructive pulmonary disease or chronic cough. Next to prominent neuropeptides such as tachykinins or vasoactive intestinal polypeptide (VIP), calcitonin gene-related peptide (CGRP) has long been suggested to participate in airway physiology and pathophysiology. CGRP is a 37 amino-acid peptide which is expressed by nerve fibers projecting to the airways and by pulmonary neuroendocrine cells. The most prominent effects of CGRP in the airways are vasodilatation and in a few instances bronchoconstriction. A further pulmonary effect of CGRP is the induction of eosinophil migration and the stimulation of beta-integrin-mediated T cell adhesion to fibronectin at the site of inflammation. By contrast, CGRP inhibits macrophage secretion and the capacity of macrophages to activate T-cells, indicating a potential anti-inflammatory effect. Due to the complex pulmonary effects of CGRP with bronchoconstriction and vasodilatation and diverse immunomodulatory actions, potential anti-asthma drugs based on this peptide have not been established so far. However, targeting the effects of CGRP may be of value for future strategies in nerve modulation. PMID- 12749829 TI - Novelties in the field of antimicrobial compounds for the treatment of lower respiratory tract infections. AB - Emergence of antimicrobial resistance is a growing problem and a public health threat. New drugs must be designed with emerging needs in mind: specific resistant and hard-to-treat organisms. But the difficulty to find real new drugs is a major problem. Only the oxazolidinones, the cationic peptides and the lipopeptide antibiotics can be truly regarded as structurally novel drugs, although the peptide deformylase inhibitors and, possibly, the pleuromutilins can be considered a potential advancement in the field. Obviously, these antibiotics must be reserved only to cases of documented ineffectiveness of the common antimicrobial agents. PMID- 12749830 TI - Tolerability of a high dose of budesonide/formoterol in a single inhaler in patients with asthma. AB - This randomised, double-blind, double-dummy, crossover, placebo-controlled study assessed the acute tolerability of budesonide/formoterol in a single inhaler (Symbicort Turbuhaler, AstraZeneca) administered as a high dose. Fourteen patients with asthma receiving budesonide/formoterol maintenance treatment (two inhalations of 160/4.5 microg twice daily) inhaled 10 additional doses of budesonide/formoterol 1600/45 microg (total daily dose including morning dose of maintenance treatment 1920/54 microg) or formoterol 45 microg (Oxis Turbuhaler, AstraZeneca; total daily dose including morning dose of maintenance treatment 54 microg formoterol) or placebo in addition to the morning dose of maintenance treatment on 3 separate study days. Serum potassium, pulse rate, blood pressure and ECG were assessed at regular intervals over a 12-h period following dosing. Blood glucose and plasma lactate were assessed over 3 h following dosing. Changes in serum potassium, pulse rate, blood pressure, QTc, blood glucose and plasma lactate occurring with budesonide/formoterol, though statistically significantly different from placebo (P<0.05), were considered clinically unimportant. No clinically relevant differences were identified between active treatments. In conclusion, budesonide/formoterol in a single inhaler is well tolerated at high doses such as might be used by patients using budesonide/formoterol for relief of symptoms of asthma. PMID- 12749831 TI - A comparison of salmeterol and formoterol in attenuating airway responses to short-acting beta2-agonists. AB - In vitro data suggest that salmeterol, contrary to formoterol, can partly antagonise the effect of short-acting beta(2)-agonist rescue medication. To explore whether this occurs in vivo, we compared the effects of increasing doses (200-3200 microg) of fenoterol on the recovery of methacholine induced bronchoconstriction as well as PD(20) methacholine in 23 asthmatic patients, during two-week treatment periods with placebo, and standard doses of salmeterol or formoterol in a double blind, double-dummy, crossover study. Salmeterol showed a slightly higher propensity for the development of bronchodilator tolerance. The recovery of methacholine induced bronchoconstriction was more complete during regular use of formoterol relative to salmeterol. During regular use of both long acting beta(2)-agonists the bronchoprotective efficacy of fenoterol was attenuated, but this was more pronounced during salmeterol than during formoterol. The mean maximum increase in PD(20) metacholine after the highest dose of fenoterol was 3.97 DD during placebo, 2.47 DD during formoterol (p<0.001) and 1.81 DD during salmeterol treatment (p<0.001). We conclude that in asthmatic patients the efficacy of short-acting beta(2)-adrenoceptor agonists can be significantly attenuated during regular use of long-acting beta(2)-agonists. In this respect, differences were observed between salmeterol and formoterol that may represent the expression of partial antagonism by salmeterol. PMID- 12749832 TI - Lung physiology and histopathology during cumulated exposure to nitric oxide in combination with assisted ventilation in healthy piglets. AB - Inhaled nitric oxide (iNO) is routinely used for hypoxic respiratory failure and persistent pulmonary hypertension of the newborn, and investigation of its new indications requiring various levels of iNO is underway. Cumulated exposure of high level of iNO may exert adverse effects on lung function and morphology, which may be confounded with ventilator-associated lung injury. Sixteen healthy piglets (5.5-11 kg) were anaesthetised, intubated and mechanically ventilated at low FiO(2) on continuous positive airway pressure and pressure support mode. The animals were randomly allocated to receive 40 ppm iNO (NO group, n=8) or no iNO (Control group, n=8). In both groups at 24 and 48 h, mild to moderate lung injury was observed, with mean values of PaO(2)/FiO(2)<300 mmHg. Compared to the Control, iNO at 40 ppm for 24-48 h did not adversely affect dynamic compliance or resistance of respiratory system, oxygenation, pulmonary and systemic hemodynamics. Neither did it affect composition and surface activity of surfactant phospholipids and white cell counts in bronchoalveolar lavage fluid. Inhaled NO resulted in elevated total serum nitrite/nitrate to 352+/-90 micromol/l and methemoglobin (MetHb) to 5.0+/-3.4%, in contrast to 88+/-38 micromol/l and 0.88+/-0.52% in the Control; 50% of the iNO animals having MetHb>3%. The lung injury scores as well as alveolar expansion were similar between the two groups at 24 h. At 48 h, low wet/dry lung weight ratio and lung injury score were found in the NO group. We conclude that no significant adverse effects on lung physiology and structure were found in the piglets receiving 40 ppm iNO for 24 or 48 h, on the contrary lung injury was moderately alleviated. The significantly impaired gas exchange over time associated with discrete morphological changes suggests adverse effects of prolonged positive pressure breathing and not necessarily exposure to oxides of nitrogen. PMID- 12749833 TI - Differential inhibition of thrombin- and EGF-stimulated human cultured airway smooth muscle proliferation by glucocorticoids. AB - The present study compared the effects of glucocorticoids on thrombin- and EGF stimulated proliferation in human cultured airway smooth muscle (ASM) to identify pathways that may be differentially regulated by glucocorticoids. Mitogenic responses to thrombin were inhibited by extracellular-regulated kinase (ERK 1/2) and phosphoinositide 3-kinase (PI3K) inhibitors, whereas mitogenic responses to EGF were inhibited by ERK 1/2 and PI3K inhibitors as well as by the p38 mitogen activated protein kinase inhibitor, SB203580 (10 microM). Mitogenic responses to thrombin were more sensitive to inhibition by dexamethasone (Dex) or fluticasone propionate (FP) than were those to EGF. Elevated cyclin D1 protein and mRNA levels induced by thrombin and EGF were attenuated equally by glucocorticoids. The protein or mRNA levels of the cyclin-dependent kinase inhibitors (cdki) p21(Cip1), p27(Kip1) were unaffected by Dex treatment of ASM cells treated with mitogens. The resistance of EGF-induced proliferation to inhibition by glucocorticoids is not associated with a failure to regulate cyclin D1 induction, nor does it appear to be explained by differential regulation of the levels of the cdki's, p21(Cip1) and p27(Kip1). PMID- 12749834 TI - Effective generation of transgenic mice by Bovine papillomavirus type 1 based self-replicating plasmid that is maintained as extrachromosomal genetic element in three generations of animals. AB - The objective of our study was to analyze the efficiency and the properties of the inheritance of the Bovine papillomavirus type 1 (BPV1) replicator-based plasmid used as vector system for generation of transgenic animals. Previously, we have characterized a series of self-replicating plasmid vectors containing all viral factors necessary and sufficient for stable extrachromosomal replication of the BPV1 genome in the tissue culture system. We also demonstrated that the designed replicating vector system has a considerable benefit in the transgene expression, if compared to the regular expression vector. The vector, which showed the highest stability and maintenance function in the tissue culture was chosen for generation of the transgenic mice by pronuclear injections of the circular supercoiled plasmid. This method resulted in successful production of transgenic animals. Transmission efficiency of the vectors into the F(1) generation of animals varied between 0 and 48%, whereas transmission into the F(2) generation was uniformly near 50%. The maintenance of the vector-plasmids in the F(2) generation of transgenic animals as extrachromosomal genetic element was demonstrated by rescue of the plasmid into the Escherichia coli. PMID- 12749835 TI - The patchwork nature of rolling-circle plasmids: comparison of six plasmids from two distinct Bacillus thuringiensis serotypes. AB - Bacillus thuringiensis, the entomopathogenic bacteria from the Bacillus cereus group, harbors numerous extrachromosomal molecules whose sizes vary from 2 to more than 200kb. Apart from the genes coding for the biopesticide delta endotoxins located on large plasmids, little information has been obtained on these plasmids and their contribution to the biology of their host. In this paper, we embarked on a detailed comparison of six small rolling-circle replicating (RCR) plasmids originating from two major B. thuringiensis strains. The complete nucleotide sequences of plasmid pGI1, pGI2, pGI3, pTX14-1, pTX14-2, and pTX14-3 have been obtained and compared. Replication functions, comprising, for each plasmid, the gene encoding the Rep-protein, double-strand origin of replication (dso), single-strand origin of replication (sso), have been identified and analyzed. Two new families, or homology groups, of RCR plasmids originated from the studies of these plasmids (Group VI based on pGI3 and Group VII based on pTX14-3). On five of the six plasmids, loci involved in conjugative mobilization (Mob-genes and origin of transfer (oriT)) were identified. Plasmids pTX14-1, pTX14-2, and pTX14-3 each harbor an ORF encoding a polypeptide containing a central domain with repetitive elements similar to eukaryotic collagen (Gly-X-Y triplets). These genes were termed bcol for Bacillus-collagen like genes. PMID- 12749836 TI - Nucleotide sequence based characterizations of two cryptic plasmids from the marine bacterium Ruegeria isolate PR1b. AB - Two plasmids, 76 and 148 kb in size, isolated from Ruegeria strain PR1b were entirely sequenced. These are the first plasmids to be characterized from this genus of marine bacteria. Sequence analysis revealed a biased distribution of function among the putative proteins encoded on the two plasmids. The smaller plasmid, designated pSD20, encodes a large number of putative proteins involved in polysaccharide biosynthesis and export. The larger plasmid, designated pSD25, primarily encodes putative proteins involved in the transport of small molecules and in DNA mobilization. Sequence analysis revealed uncommon potential replication systems on both plasmids. pSD25, the first repABC-type replicon isolated from the marine environment, actually contains two repABC-type replicons. pSD20 contains a complex replication region, including a replication origin and initiation protein similar to iteron-containing plasmids (such as pSW500 from the plant pathogen Erwinia stewartii) linked to putative RepA and RepB stabilization proteins of a repABC-type replicon and is highly homologous to a plasmid from the phototrophic bacterium Rhodobacter sphaeroides. Given the nature of the putative proteins encoded by both plasmids it is possible that these plasmids enhance the metabolic and physiological flexibility of the host bacterium, and thus its adaptation to the marine sediment environment. PMID- 12749837 TI - Three new insertion sequence elements ISLdl2, ISLdl3, and ISLdl4 in Lactobacillus delbrueckii: isolation, molecular characterization, and potential use for strain identification. AB - A group of new insertion sequence (IS) elements, ISLdl2, ISLdl3, and ISLdl4, from Lactobacillus delbrueckii subsp. lactis ATCC 15808 was isolated, characterized, and used for strain identification together with ISLdl1, recently characterized as an L. delbrueckii IS element belonging to the ISL3 family. ISLdl2 was 1367 bp in size and had a 24 bp IR and an 8 bp DR. The single ORF of ISLdl2 encoded a protein of 392 aa similar to transposases of the IS256 family. ISLdl3 had a single ORF encoding a protein of 343 aa similar to transposases of the IS30 family. Finally, ISLdl4 had a single ORF encoding a protein of 406 aa and displayed homology to the transposases of the IS110 family. ISLdl4 was only slight different from ISL4 (Accession No. AY040213). ISLdl1, ISLdl2, and ISLdl4 were present in all of the 10 L. delbrueckii subsp. lactis and subsp. delbrueckii strains tested, as well as in three of the 11 L. delbrueckii subsp. bulgaricus strains tested. ISLdl3 was present only in four closely related strains of L. delbrueckii subsp. lactis. These IS elements were not observed in Lactobacillus rhamnosus, Lactobacillus acidophilus, Lactobacillus helveticus, or Lactobacillus plantarum. A cluster of IS elements, ISLdl1, ISLdl2, ISLdl3, ISLdl4, and ISL6, was observed in L. delbrueckii subsp. lactis strain ATCC 15808. Within this cluster, ISLdl4 was inserted into ISLdl1 between the left IR and the start codon of ORF455, encoding a putative transposase. Most of the integration sites of the IS elements were strain-specific. We have observed that IS elements can migrate from one strain to another as integral parts of bacterial DNA by using phage LL-H as a vehicle. We demonstrate for the first time that inverse PCR and vectorette PCR methods with primers based on sequences of the IS elements could be used for identification of L. delbrueckii strains. PMID- 12749839 TI - Construction of the mobilizable plasmid pMV158GFP, a derivative of pMV158 that carries the gene encoding the green fluorescent protein. AB - Plasmid pMV158 has been employed to construct cloning non-mobilizable vectors for various Gram-positive organisms. Here we report the construction of a mobilizable pMV158-based plasmid that harbors the gene encoding the green fluorescent protein under the control of a promoter inducible by maltose. The plasmid was mobilized between strains of Streptococcus pneumoniae as well as from S. pneumoniae to Lactococcus lactis or Enterococcus faecalis at the same frequency as its parental. Transconjugant that received the GFP-tagged plasmid could be detected by their fluorescence, which was especially high in E. faecalis cells. PMID- 12749838 TI - The replicon theory 40 years: an EMBO workshop held in Villefranche sur Mer, France, January 18-23, 2003. AB - It is now 40 years since Jacob, Brenner, and Cuzin presented their Replicon Theory at a Cold Spring Harbor Symposium. The theory was based on their fundamental studies of the sexual system of Escherichia coli which led to the realisation that only specific sequences are able to replicate. They introduced the concept of a replicon consisting of a replicator (a DNA sequence) and a structural gene for an initiator protein. They also proposed a model for how replication of the bacterial chromosome might fit into the bacterial cell cycle. To commemorate the anniversary, an EMBO Workshop was organised in Villefranche on the Riviera of France. During the Workshop, the state of the art of cell-cycle studies of prokaryotic and eukaryotic organisms was presented and discussed in the presence of two of the fathers of the Replicon Theory, Jacob and Cuzin. PMID- 12749840 TI - Metabolic flux analysis of hepatocyte function in hormone- and amino acid supplemented plasma. AB - Understanding the metabolic and regulatory pathways of hepatocytes is important for biotechnological applications involving liver cells. Previous attempts to culture hepatocytes in plasma yielded poor functional results. Recently we reported that hormone (insulin and hydrocortisone) and amino acid supplementation reduces intracellular lipid accumulation and restores liver-specific function in hepatocytes exposed to heparinized human plasma. In the current study, we performed metabolic flux analysis (MFA) using a simplified metabolic network model of cultured hepatocytes to quantitively estimate the changes in lipid metabolism and relevant intracellular pathways in response to hormone and amino acid supplementation. The model accounts for the majority of central carbon and nitrogen metabolism, and assumes pseudo-steady-state with no metabolic futile cycles. We found that beta-oxidation and tricarboxylic acid (TCA) cycle fluxes were upregulated by both hormone and amino acid supplementation, thus enhancing the rate of lipid oxidation. Concomitantly, hormone and amino acid supplementation increased gluconeogenic fluxes. This, together with an increased rate of glucose clearance, caused an increase in predicted glycogen synthesis. Urea synthesis was primarily derived from ammonia and aspartate generated through transamination reactions, while exogenous ammonia removal accounted for only 3-6% of the urea nitrogen. Amino acid supplementation increased the endogenous synthesis of oxaloacetate, and in turn that of aspartate, a necessary substrate for the urea cycle. These findings from MFA provide cues as to which genes/pathways relevant to fatty acid oxidation, urea production, and gluconeogenesis may be upregulated by plasma supplementation, and are consistent with current knowledge of hepatic amino acid metabolism, which provides further credence to this approach for evaluating the metabolic state of hepatocytes under various environmental conditions. PMID- 12749841 TI - Metabolic flux analysis of xylose metabolism in recombinant Saccharomyces cerevisiae using continuous culture. AB - This study focused on elucidating metabolism of xylose in a Saccharomyces cerevisiae strain that overexpresses xylose reductase and xylitol dehydrogenase from Pichia stipitis, as well as the endogenous xylulokinase. The influence of xylose on overall metabolism was examined supplemented with low glucose levels with emphasis on two potential bottlenecks; cofactor requirements and xylose uptake. Results of metabolic flux analysis in continuous cultivations show changes in central metabolism due to the cofactor imbalance imposed by the two step oxidoreductase reaction of xylose to xylulose. A comparison between cultivations on 27:3g/L xylose-glucose mixture and 10g/L glucose revealed that the NADPH-generating flux from glucose-6-phosphate to ribulose-5-phosphate was almost tenfold higher on xylose-glucose mixture and due to the loss of carbon in that pathway the total flux to pyruvate was only around 60% of that on glucose. As a consequence also the fluxes in the citric acid cycle were reduced to around 60%. As the glucose level was decreased to 0.1g/L the fluxes to pyruvate and in the citric acid cycle were further reduced to 30% and 20%, respectively. The results from in vitro and in vivo xylose uptake measurements showed that the specific xylose uptake rate was highest at the lowest glucose level, 0.1g/L. PMID- 12749842 TI - Engineering metabolism and product formation in Corynebacterium glutamicum by coordinated gene overexpression. AB - Single gene overexpression in product pathways such as lysine synthesis has often been employed in metabolic engineering efforts aiming at pathway flux amplification and metabolite overproduction. This approach is limited due to metabolic flux imbalances that often lead to unpredictable physiological responses and suboptimal metabolite productivity. This deficiency can be overcome by the coordinated overexpression of more than one flux controlling genes in a production pathway selected by considering their individual contributions on the cell physiology This concept is demonstrated by the simultaneous overexpression of pyruvate carboxylase and aspartate kinase, two key enzymes in central carbon metabolism and the lysine production pathway in Corynebacterium glutamicum. Contrary to expectations based on the importance of each of these two genes in lysine production, the monocistronic overexpression of either gene results in marginal changes in the overall lysine productivity due to either reduced cell growth or reduced lysine specific productivity. In contrast, the simultaneous amplification of the activities of the two enzymes yielded more than 250% increase of the lysine specific productivity in lactate minimal medium without affecting the growth rate or final cell density of the culture. These results demonstrate that significant flux amplification in complex pathways involving central carbon metabolism is possible through coordinated overexpression of more than one gene in the pathway. This can be achieved either by external, gene expression inducing, controls or controls responding to the physiological cellular state. PMID- 12749843 TI - Influence of the adipate and dissolved oxygen concentrations on the beta-lactam production during continuous cultivations of a Penicillium chrysogenum strain expressing the expandase gene from Streptomyces clavuligerus. AB - The influence of adipate concentration and dissolved oxygen on production of adipoyl-7-aminodeacetoxycephalosporanic acid (ad-7-ADCA) by a recombinant strain of Penicillium chrysogenum expressing the expandase gene from Streptomyces clavuligerus was studied in glucose-limited continuous cultures. Operating conditions were maintained constant but the adipate and dissolved oxygen concentrations (DOC) were varied separately in a range from 1 to 37.5gl(-1) and from 2% to 125% air saturation (%AS), respectively. The total beta-lactams specific productivity, r(ptotal), was not significantly changed for adipate concentrations from 5 to 25gl(-1), but the flux towards an unknown by-product decreased as the adipate concentration increased. Investigations at different DOC showed that r(ptotal) was stable around 18 micro molgDW(-1)h(-1) for DOC being in the range from 15 to 125%AS. When DOC was decreased from 15 to 7%AS, r(ptotal) increased to 25 micro molgDW(-1)h(-1), mainly due to a two-fold increase in the adipoyl-6-aminopenicillanic acid (ad-6-APA) specific productivity. PMID- 12749844 TI - Reducing maintenance metabolism by metabolic engineering of respiration improves riboflavin production by Bacillus subtilis. AB - We present redirection of electron flow to more efficient proton pumping branches within respiratory chains as a generally applicable metabolic engineering strategy, which tailors microbial metabolism to the specific requirements of high cell density processes by improving product and biomass yields. For the example of riboflavin production by Bacillus subtilis, we reduced the rate of maintenance metabolism by about 40% in a cytochrome bd oxidase knockout mutant. Since the putative Yth and the caa(3) oxidases were of minor importance, the most likely explanation for this improvement is translocation of two protons per transported electron via the remaining cytochrome aa(3) oxidase, instead of only one proton via the bd oxidase. The reduction of maintenance metabolism, in turn, significantly improved the yield of recombinant riboflavin and B. subtilis biomass in fed-batch cultures. PMID- 12749845 TI - Metabolic engineering of beta-lactam production. AB - Metabolic engineering has become a rational alternative to classical strain improvement in optimisation of beta-lactam production. In metabolic engineering directed genetic modification are introduced to improve the cellular properties of the production strains. This has resulted in substantial increases in the existing beta-lactam production processes. Furthermore, pathway extension, by heterologous expression of novel genes in well-characterised strains, has led to introduction of new fermentation processes that replace environmentally damaging chemical methods. This minireview discusses the recent developments in metabolic engineering and the applications of this approach for improving beta-lactam production. PMID- 12749847 TI - Phosphorylation-dependent structural alterations in the small hsp30 chaperone are associated with cellular recovery. AB - Small heat shock proteins (hsps) act as molecular chaperones by preventing the thermal aggregation and unfolding of cellular protein; however, the manner by which cells regulate chaperone activity remains unclear. In the present study, we examined the role of phosphorylation on the chaperone function of the Xenopus small hsp30. Both heat stress and sodium arsenite treatment in A6 cells resulted in a rapid activation of p38alpha and MAPKAPK-2. Surprisingly, the association of MAPKAPK-2 with hsp30 and its subsequent phosphorylation were more prevalent during recovery after heat stress. Treatment of A6 cells with SB203580, an inhibitor of the p38 MAP kinase pathway, resulted in a loss of hsp30 phosphorylation. Phosphorylation resulted in the formation of smaller multimeric hsp30 complexes and resulted in a significant loss of secondary structure. Consequently the phosphorylation-induced structural changes severely compromised the ability of hsp30 to prevent the heat-induced aggregation of citrate synthase and luciferase in vitro. We confirmed that the loss of chaperone activity was coincident with an attenuated binding of phosphorylated hsp30 with target proteins. Our data suggest that phosphorylation may be necessary to regulate the post-heat stress molecular chaperone activity of hsp30. PMID- 12749846 TI - Epidermal growth factor increased the expression of alpha2beta1-integrin and modulated integrin-mediated signaling in human cervical adenocarcinoma cells. AB - Epidermal growth factor (EGF) receptor (EGFR) is involved in various basic biochemical pathways and is thus thought to play an important role in cell migration. We examined the effect of EGF on motility, migration, and morphology of a human adenocarcinoma cell line CAC-1. EGF treatment increased the motility of cervical adenocarcinoma cells and promoted migration of the cells on fibronectin and type IV collagen. EGF induced morphological changes with lamellipodia during EGFR-mediated motility. The results of an immunoprecipitation study showed that EGF up-regulated the expression of alpha2beta1-integrin in a dose-dependent manner. EGF-induced cell migration was blocked by alpha2beta1 integrin antibody. Our results also showed that EGF treatment stimulated the level of tyrosine dephosphorylation of FAK, which is required for EGF-induced changes in motility, migration, and cell morphology. A tyrosine kinase inhibitor (ZD1839) blocked EGF-induced changes in cervical adenocarcinoma cells. The results suggest that EGF promotes cell motility and migration and increases the expression of alpha2beta1-integrin, possibly by decreasing FAK phosphorylation. PMID- 12749848 TI - Expression of Smac/DIABLO in ovarian carcinoma cells induces apoptosis via a caspase-9-mediated pathway. AB - We have constructed Ad CMV-Smac, a recombinant adenovirus encoding Smac/DIABLO, the recently described second mitochondrial activator of caspases. Transfection of ovarian carcinoma cells with Ad CMV-Smac at multiplicities of infection of 3 60 pfu/cell leads to increasing apoptosis in a dose-dependent manner. Western blot analysis confirms that Smac-induced apoptosis proceeds via a pathway mediated primarily by caspase-9 that can be inhibited by zLEHD-fmk and overexpression of the X-linked inhibitor of apoptosis protein (XIAP). In contrast, there is no cleavage of either caspase-8 or caspase-12. Ad CMV-Smac appears to induce apoptosis independently of cytochrome c release from mitochondria and is not inhibited by overexpression of Bcl-2. Ad CMV-Smac can combine with other proapoptotic factors, such as cisplatin, paclitaxel, and procaspase-3, to produce greater levels of apoptosis in transfected cells. PMID- 12749849 TI - Expression of GPI-80, a beta2-integrin-associated glycosylphosphatidylinositol anchored protein, requires neutrophil differentiation with dimethyl sulfoxide in HL-60 cells. AB - GPI-80 is a member of the amidohydrolase family that has been proposed as a potential regulator of beta2-integrin-dependent leukocyte adhesion. GPI-80 is expressed mainly in human neutrophils. Our previous studies suggested that GPI-80 expression might be associated with myeloid differentiation. To verify this, we examined whether GPI-80 is expressed on the human promyelocytic leukemia cell line HL-60 following treatment with differentiation inducers. GPI-80 expression was induced in cells treated with dimethyl sulfoxide (DMSO) to stimulate differentiation down the neutrophil pathway. On the other hand, all-trans retinoic acid (ATRA), another neutrophil-inducing reagent, induced no clear GPI 80 expression. Potent monocyte-inducing reagents such as 1alpha,25 dihydroxyvitamin D(3) or phorbol 12-myristate 13-acetate also had no significant effect on the protein expression. GPI-80-positive cells were found in the well differentiated CD11b-positive and transferrin-receptor-negative cell population. Granulocyte colony-stimulating factor, which augments neutrophil differentiation of HL-60 cells, up-regulated GPI-80 expression in the presence of DMSO. Granulocyte/macrophage colony-stimulating factor, which is known to suppress the neutrophil maturation of cells, inhibited expression. Adhesion of DMSO-induced cells was regulated by anti-GPI-80 monoclonal antibody, similar to the regulation observed in neutrophils. These results suggest that use of DMSO to induce neutrophil differentiation provides suitable conditions for GPI-80 expression, and that this culture system may be a helpful model for further study of the regulation of GPI-80 expression during myeloid differentiation. PMID- 12749850 TI - Reconstitution of TIMP-2 expression in SH-SY5Y neuroblastoma cells by 5 azacytidine is mediated transcriptionally by NF-Y through an inverted CCAAT site. AB - Advanced stage neuroblastomas (NB) exhibit a tissue inhibitor of metalloproteinase (TIMP)-2/matrix metalloproteinase (MMP) imbalance, considered a prerequisite for MMP involvement in tumor progression in vivo. Human SH-SY5Y NB cells exhibit a similar TIMP-2/MMP imbalance that promotes in vitro invasive behavior that is inhibited by exogenous TIMP-2. The DNA methyltransferase inhibitor 5-azacytidine (5-AzaC) redresses this TIMP-2/MMP imbalance, reconstituting TIMP-2 expression, without effecting that of MMP-2, by stimulating TIMP-2 transcription and inhibiting in vitro invasivity of SH-SY5Y cells. 5-AzaC stimulated transcription from a nonmethylated TIMP-2 promoter reporter gene construct consistent with regulation of a TIMP-2 transactivator. Promoter deletion and point-mutation analysis localized this effect to an inverted CCAAT element at position -73. This element bound specific complexes containing NF-YA and NF-YB proteins in SH-SY5Y nuclear extracts, the binding of which was augmented by 5-AzaC in association with enhanced levels of NF-YB protein and the function of which was confirmed by inhibition using dominant-negative NF-YA. The data highlight a novel indirect methylation-mediated mechanism for regulating the TIMP/MMP equilibrium in NB cells, involving repression of TIMP-2 relative to MMP 2 expression, dependent upon suboptimal NF-Y transcription factor function, which can be reversed by methyltransferase inhibition. PMID- 12749851 TI - Syndecan-1-mediated cell spreading requires signaling by alphavbeta3 integrins in human breast carcinoma cells. AB - Syndecans are cell surface heparan sulfate proteoglycans with regulatory roles in cell adhesion, proliferation, and differentiation [Annu. Rev. Biochem. 68 (1999) 729]. While the syndecan heparan sulfate chains are essential for matrix binding, less is known about the signaling role of their core proteins. To mimic syndecan specific adhesion, MDA-MB-231 mammary carcinoma cells were plated on antibodies against syndecan-4 or syndecan-1. While cells adherent via syndecan-4 spread, cells adherent via syndecan-1 do not. However, cells adherent via syndecan-1 can be induced to spread by Mn(2+), suggesting that activation of a beta(1) or beta(3) integrin partner is required. Surprisingly, pretreatment of cells with a function-activating beta(1) antibody does not induce spreading, whereas function blocking beta(1) integrin antibodies do, suggesting involvement of a beta(1)-to beta(3) integrin cross-talk. Indeed, blockade of beta(1) integrin activation induces alpha(v)beta(3) integrin activation detectable by soluble fibrinogen binding. Spreading in response to syndecan-1 is independent of integrin-ligand binding. Furthermore, competition with soluble murine syndecan-1 ectodomain, which does not disrupt cell adhesion, nonetheless blocks the spreading mechanism. These data suggest that the ectodomain of the syndecan-1 core protein directly participates in the formation of a signaling complex that signals in cooperation with alpha(v)beta(3) integrins; signaling via this complex is negatively regulated by beta(1) integrins. PMID- 12749852 TI - Identification of alpha-tubulin as an hsp105alpha-binding protein by the yeast two-hybrid system. AB - Hsp105alpha is a mammalian stress protein that belongs to the HSP105/110 family. Hsp105alpha prevents stress-induced apoptosis in neuronal cells and binds to Hsp70/Hsc70 and suppresses the Hsp70 chaperone activity in vitro. In this study, to further elucidate the function of Hsp105alpha, we searched for Hsp105alpha binding proteins by screening a mouse FM3A cell cDNA library with full-length Hsp105alpha using the yeast two-hybrid system and obtained alpha-tubulin as an Hsp105alpha-binding protein. Hsp105alpha bound directly to alpha-tubulin both in vitro and in vivo. Indirect immunofluorescence analysis with anti-Hsp105 and anti alpha-tubulin antibodies indicated that Hsp105alpha was colocalized with microtubules. Furthermore, the disorganization of microtubules induced by heat shock was prevented in Hsp105alpha-overexpressing COS-7 cells. These findings suggested that Hsp105alpha associates with alpha-tubulin and microtubules in cells and plays a role in protection of microtubules under conditions of stress. PMID- 12749853 TI - Upregulation and antiapoptotic role of endogenous Alzheimer amyloid precursor protein in dorsal root ganglion neurons. AB - The amyloid precursor protein (APP) is a transmembrane protein whose abnormal processing is associated with the pathogenesis of Alzheimer's disease. In this study, we examined the expression and role of cell-associated APP in primary dorsal root ganglion (DRG) neurons. When dissociated DRG cells prepared from mouse embryos were treated with nerve growth factor (NGF), neuronal APP levels were transiently elevated. DRG neurons treated with an antibody against cell surface APP failed to mature and underwent apoptosis. When NGF was withdrawn from the cultures after a 36-h NGF treatment, virtually all neurons underwent apoptosis by 48 h. During the course of apoptosis, some neurons with intact morphology contained increased levels of APP immunoreactivity, whereas the APP levels were greatly reduced in apoptotic neurons. Furthermore, affected neurons contained immunoreactivities for activated caspase-3, a caspase-cleaved APP fragment (APPDeltaC31), and Abeta. Downregulation of endogenous APP expression by treatment with an APP antisense oligodeoxynucleotide significantly increased the number of apoptotic neurons in NGF-deprived DRG cultures. Furthermore, overexpression of APP by adenovirus vector-mediated gene transfer reduced the number of apoptotic neurons deprived of NGF. These results suggest that endogenous APP is upregulated to exert an antiapoptotic effect on neurotrophin deprived DRG neurons and subsequently undergoes caspase-dependent proteolysis. PMID- 12749854 TI - Monoclonal antibody against dnmt1 arrests the cell division of xenopus early stage embryos. AB - DNA methylation plays a crucial role in embryogenesis, and Dnmt1 is known to be a key enzyme in the maintenance of DNA methylation. Dnmt1 is highly accumulated in mature oocytes and eggs. To analyze the function of the maternally accumulated Dnmt1, we injected monoclonal antibodies that specifically recognize the amino terminus of Xenopus Dnmt1 into Xenopus laevis embryos. The monoclonal antibodies inhibited the cell division of the embryos before the midblastula transition. Monoclonal antibody neither inhibited DNA methylation activity of Dnmt1 in vitro nor affected its stability in embryos. In addition, injection of alpha-amanitin, an inhibitor of transcription, did not rescue the cell division arrest. The results suggest that the inhibition of cell division by monoclonal antibodies was due neither to the direct inhibition of DNA methylation activity of Dnmt1 nor to aberrant transcription before the midblastula transition. The morphology of chromatin of the arrested cells showed that the cell cycle was arrested at interphase. This was supported by the biochemical analysis in which the arrested cells demonstrated low histone H1 kinase activity, which indicated that the cells had not entered M phase. Dnmt1 may have an important function other than DNA methylation activity for early embryogenesis in Xenopus laevis. PMID- 12749855 TI - Mechanisms involved in the inhibition of myoblast proliferation and differentiation by myostatin. AB - Muscle growth results from a set of complex processes including myogenic transcription factor's expression and activity, cell cycle withdrawal, myoblast fusion in myotubes, and acquisition of an apoptosis-resistant phenotype. Myostatin, a member of the TGFbeta family, described as a strong regulator of myogenesis in vivo Nature 387 (1997), 83; FEBS Lett. 474 (2000), 71 is upregulated during in vitro differentiation Biochem. Biophys. Res. Commun. 280 (2001), 561. To improve characterization of myostatin's myogenic influence, we stably transfected vectors expressing myostatin and myostatin antisense in C2C12 myoblasts. Here, we found that myostatin inhibits cell proliferation and differentiation. Our results also indicate that myogenin is an important target of myostatin. In addition, overexpressed but not endogenous myostatin decreases MyoD protein levels and induces changes in its phosphorylation pattern. We also established that myostatin overexpression reduces the frequency of G0/G1-arrested cells during differentiation. Conversely, inhibition of myostatin synthesis leads to enhanced cell cycle withdrawal and consequently stimulates myoblast differentiation. We examined the expression patterns of the pRb, E2F1, p53, and p21 proteins involved in cell cycle withdrawal. We found that myostatin overexpression increases p21 and p53 expression, as it does accumulation of hypophosphorylated Rb. Interestingly, myostatin overexpression strongly reduced low-mitogen-induced apoptosis, whereas antisense expression induced contrary changes. In conclusion, these data show the influence of overexpressed myostatin on myoblast proliferation, differentiation, and apoptosis is extended to endogenous myostatin. Though some differences in overexpression or inhibition of endogenous myostatin were observed, it appears that myogenin and p21 are essential targets of this growth factor. PMID- 12749856 TI - Nucleocytoplasmic shuttling of antigen in mammalian cells conferred by a soluble versus insoluble single-chain antibody fragment equipped with import/export signals. AB - The ectopic expression of antibody fragments within mammalian cells is a challenging approach for interfering with or even blocking the biological function of the intracellular target. For this purpose, single-chain Fv (scFv) fragments are generally preferred. Here, by transfecting several mammalian cell lines, we compared the intracellular behavior of two scFvs (13R4 and 1F4) that strongly differ in their requirement of disulphide bonding for the formation of active molecules in bacteria. The scFv 13R4, which is correctly folded in the bacterial cytoplasm, was solubly expressed in all cell lines tested and was distributed in their cytoplasm and nucleus, as well. In addition, by appending to the 13R4 molecules the SV40 T-antigen nuclear localisation signal (NLS) tag, cytoplasmic-coexpressed antigen was efficiently retargeted to the nucleus. Compared to the scFv 13R4, the scFv 1F4, which needs to be secreted in bacteria for activity, accumulated, even with the NLS tag, as insoluble aggregates within the cytoplasm of the transfected cells, thereby severely disturbing fundamental functions of cell physiology. Furthermore, by replacing the NLS tag with a leucine-rich nuclear export signal (NES), the scFv 13R4 was exclusively located in the cytoplasm, whereas the similarly modified scFv 1F4 still promoted cell death. Coexpression of NES-tagged 13R4 fragments with nuclear antigen promoted its efficient retargeting to the cytoplasm. This dominant effect of the NES tag was also observed after exchange of the nuclear signals between the scFv 13R4 and its antigen. Taken together, the results indicate that scFvs that are active in the cytoplasm of bacteria may behave similarly in mammalian cells and that the requirement of their conserved disulphide bridges for activity is a limiting factor for mediating the nuclear import/export of target in a mammalian cell context. The described shuttling effect of antigen conferred by a soluble scFv may represent the basis of a reliable in vivo assay of effective protein- protein interactions. PMID- 12749858 TI - Elevated expression of exogenous Rad51 leads to identical increases in gene targeting frequency in murine embryonic stem (ES) cells with both functional and dysfunctional p53 genes. AB - The Rad51 gene is the mammalian homologue of the bacterial RecA gene and catalyses homologous recombination in mammalian cells. In some cell types Rad51 has been shown to interact with p53, leading to inhibition of Rad51 activity. Here, we show a two- to four-fold increase in gene-targeting frequency at the HPRT locus using murine ES clones preengineered to overexpress Rad51, and a twofold increase in targeting frequency when a Rad51 expression cassette was cointroduced to wild-type ES cells with the targeting construct. In addition to its effect on homologous recombination, we show that Rad51 may down-regulate illegitimate recombination. We investigated the dependence of these phenomena upon p53 and found no evidence that the Rad 51-mediated increase is affected by the functional status of p53, a conclusion supported by the observed cytoplasmic localisation of p53 in ES cells following electroporation. Furthermore, in the absence of additional Rad51, p53-deficient ES cells do not have elevated rates of homologous recombination with extrachromosomal DNA. These findings demonstrate that Rad51 levels modify both homologous and illegitimate recombination, but that these phenomena are independent of p53 status. PMID- 12749857 TI - Hepatocyte nuclear factor (HNF)-4alpha triggers formation of functional tight junctions and establishment of polarized epithelial morphology in F9 embryonal carcinoma cells. AB - F9 murine embryonal carcinoma cells provide an attractive system for facilitating molecular mechanisms for epithelial morphogenesis, since they have the capability of differentiating into polarized epithelial cells bearing an apical junctional complexes. We previously showed that a specific retinoid X receptor-retinoic acid receptor heterodimer transduced retinoid signals for biogenesis of functional tight junctions in F9 cells (Exp. Cell Res. 263, (2001) 163). In the present study we generated F9 cells expressing doxycycline-inducible hepatocyte nuclear factor (HNF)-4alpha, a nuclear receptor. We herein show that induction of HNF 4alpha initiates differentiation of F9 cells to polarized epithelial cells, in which tight-junction proteins occludin, claudin-6, claudin-7, and ZO-1 are concentrated at the apical-most regions of lateral membranes. Expression of occludin, claudin-6, and claudin-7 was induced in the cells by doxycycline treatment in a dose- and time-dependent manner, in terms of the amount of HNF 4alpha. In contrast, expression levels of ZO-1, ZO-2, E-cadherin, and beta catenin were not altered by HNF-4alpha. We also demonstrate, by analysis of diffusion of labeled sphingomyelin, that the fence function of tight junctions is achieved by induction of HNF-4alpha. These findings indicate that HNF-4alpha triggers de novo formation of functional tight junctions and establishment of epithelial cell polarity. PMID- 12749860 TI - Ratjadones inhibit nuclear export by blocking CRM1/exportin 1. AB - In addition to previously isolated ratjadone A we describe three new members of this family, ratjadones B, C, and D, from another strain of the myxobacterium Sorangium cellulosum. We have investigated the properties of these ratjadones with respect to their activity on mammalian cell lines. We found IC(50) values in the picomolar range and a significant increase in the size of nuclei. A further examination showed that they inhibit the export of the leucine-rich nuclear export signal (LR-NES) containing proteins in different cell lines. Ratjadones are able to inhibit the formation of the nuclear export complex composed of the CRM1, RanGTP, and the cargo protein, as shown by two different in vitro assays. Finally, the binding of ratjadone C to CRM1 was demonstrated. These ratjadone activities are in the same concentration range as described for the polyketide leptomycin B (LMB) from Streptomyces sp. Like LMB, it seems that the ratjadones covalently bind to CRM1, inhibit cargo protein binding via LR-NES, and thereby block nuclear export. Thus, the ratjadones represent a new class of natural compounds which inhibit proliferation in eukaryotes by blocking nuclear export. PMID- 12749859 TI - Mouse NIPK interacts with ATF4 and affects its transcriptional activity. AB - Neuronal cell death-inducible putative kinase (NIPK) is a protein with an unknown function encoded by a gene activated in neuronal cells in cell death-causing conditions (disruption of calcium homeostasis, trophic factor deprivation). Using the yeast two-hybrid screening of an embryonic mouse cDNA library, we identified activating transcription factor 4 (ATF4) as a protein binding to mouse (m) NIPK. The critical domain for mNIPK-binding resides in a 72 amino acid stretch near the N-terminus of ATF4, covering the second leucine zipper motif and the preceding region. mNIPK expressed as fusion protein with enhanced yellow fluorescence protein (EYFP) is localized predominantly in the nucleus, and the mNIPK-ATF4 complex can be immunoprecipitated from cells cotransfected with epitope-tagged mNIPK and ATF4 constructs. The expression of both mNIPK and ATF4 is upregulated in the neuronal cell line GT1-7 in response to disruption of calcium homeostasis by thapsigargin, but ATF4 is induced more rapidly than mNIPK. The coexpression of mNIPK inhibits ATF4 CRE-dependent transcriptional activation activity in transiently transfected cells. At the same time, ATF4 degradation rate is not increased in the cells coexpressing mNIPK, and ATF4, associated to mNIPK, is able to bind to CRE. Thus, mNIPK is a novel regulator of ATF4 transcriptional activity. PMID- 12749861 TI - Nuclear poly(A)-binding protein PABPN1 is associated with RNA polymerase II during transcription and accompanies the released transcript to the nuclear pore. AB - The nuclear poly(A)-binding protein, PABPN1, has been previously shown to regulate mRNA poly(A) tail length and to interact with selected proteins involved in mRNA synthesis and trafficking. To further understand the role of PABPN1 in mRNA metabolism, we used cryo-immunoelectron microscopy to determine the fate of PABPN1 at various stages in the assembly and transport of the Chironomus tentans salivary gland Balbiani ring (BR) mRNA ribonucleoprotein (mRNP) complex. PABPN1 is found on BR mRNPs within the nucleoplasm as well as on mRNPs docked at the nuclear pore. Very little PABPN1 is detected on the cytoplasmic side of the nuclear envelope, suggesting that PABPN1 is displaced from mRNPs during or shortly after passage through the nuclear pore. Surprisingly, we also find PABPN1 associated with RNA polymerase II along the chromatin axis of the BR gene. Our results suggest that PABPN1 binds to the polymerase before, at, or shortly after the start of transcription, and that the assembly of PABPN1 onto the poly(A) tail may be coupled to transcription. Furthermore, PABPN1 remains associated with the released BR mRNP until the mRNP is translocated from the nucleus to the cytoplasm. PMID- 12749863 TI - Effects of cadmium on DNA-(Cytosine-5) methyltransferase activity and DNA methylation status during cadmium-induced cellular transformation. AB - Cadmium is a human carcinogen that likely acts via epigenetic mechanisms. Since DNA methylation alterations represent an important epigenetic event linked to cancer, the effect of cadmium on DNA methyltransferase (MeTase) activity was examined using in vitro (TRL1215 rat liver cells) and ex vivo (M.SssI DNA MeTase) systems. Cadmium effectively inhibited DNA MeTases in a manner that was noncompetitive with respect to substrate (DNA), indicating an interaction with the DNA binding domain rather than the active site. Based on these results, the effects of prolonged cadmium exposure on DNA MeTase and genomic DNA methylation in TRL1215 cells were studied. After 1 week of exposure to 0-2.5 microM cadmium, DNA MeTase activity was reduced (up to 40%) in a concentration-dependent fashion, while genomic DNA methylation showed slight but significant reductions at the two highest concentrations. After 10 weeks of exposure, the cells exhibited indications of transformation, including hyperproliferation, increased invasiveness, and decreased serum dependence. Unexpectedly, these cadmium transformed cells exhibited significant increases in DNA methylation and DNA MeTase activity. These results indicate that, while cadmium is an effective inhibitor of DNA MeTase and initially induces DNA hypomethylation, prolonged exposure results in DNA hypermethylation and enhanced DNA MeTase activity. PMID- 12749862 TI - Mycoplasma arthritidis superantigen (MAM)-induced macrophage nitric oxide release is MHC class II restricted, interferongamma dependent, and toll-like receptor 4 independent. AB - Mycoplasma arthritidis causes arthritis in rodents that resembles human rheumatoid arthritis. It produces a superantigen (MAM) that stimulates production of cytokines by making a bridge between lymphocyte T-cell receptor with the appropriate Vbeta chain, and H-2 1-Ealpha MHC class II molecules. Here we studied MAM-induced nitric oxide (NO) production in mouse peritoneal macrophages and found that it was: (1) time and concentration dependent, (2) possibly derived from inducible NOS synthase since it was reduced significantly by amino guanidine pretreatment, (3) restricted to H-2(K) (C3H/HePas and C3H/HeJ) and H-2(d) strains (BALB/c), (4) independent of TLR4 signaling since the coisogenic strains C3H/HePas and C3H/HeJ (TLR4 deficient) produced similar levels of NO following MAM stimulation, (5) potentiated by lipopolysaccharide, and (6) dependent on the presence of nonadherent peritoneal cells. Neutralization of interferon-gamma (IFNgamma in the peritoneal cell cultures with monoclonal antibodies abolished MAM-induced NO production. Addition of rIFNgamma to the adherent cells substituted the nonadherent cells for MAM-induced NO production. A macrophage cell line, J774A.1 (H-2(d)), also produced NO upon MAM stimulation but only when BALB/c spleen lymphocytes were added. Thus, in murine macrophages, MAM induces NO production that is dependent on signaling through MHC class II molecules and IFNgamma but independent of TLR4 expression. PMID- 12749864 TI - Differential binding to dorsal and ventral cell surfaces of fibroblasts: effect on collagen phagocytosis. AB - Matrix remodeling by phagocytic fibroblasts is essential for growth and development but the regulatory processes are undefined. We evaluated the impact of spreading on the binding step of collagen phagocytosis with a novel culture system that more closely replicates phagocytosis in vivo than previous models. 3T3 cells were plated on collagen-coated beads, thereby loading only ventral surfaces (adhesion with spreading), or were allowed to spread on collagen films and then loaded with beads on their dorsal surfaces (adhesion without spreading). Ventral surfaces bound three-fold more beads than dorsal surfaces which was accompanied by accelerated phagosomal maturation. Arp3 and cortactin, markers of the actin-associated spreading machinery, strongly accumulated around ventrally but not dorsally loaded beads, suggesting that spreading contributes to enhanced binding of ventral surfaces. Further, ventral surfaces exhibited two-fold more free alpha2beta1 integrins, the major collagen receptors. Notably, compared to cells spread on collagen substrates, spreading cells exhibited a three-fold higher alpha2beta1 mobile fraction which was correlated with limited engagement of ventral receptors by actin filaments. Thus integrin ligation by actin filaments regulates the mobility of collagen receptors which in turn mediates the enhanced binding of collagen beads on spreading surfaces. PMID- 12749865 TI - Cisplatin triggers apoptotic or nonapoptotic cell death in Fanconi anemia lymphoblasts in a concentration-dependent manner. AB - Cells derived from Fanconi anemia (FA) patients are hypersensitive for cross linking agents, such as cisplatin, that are potent inducers of programmed cell death (PCD). Here, we studied cisplatin hypersensitivity in FA in relation to the mechanism of PCD in lymphoblastoid cells representing FA groups A and C. In FA cells, a low concentration of cisplatin caused chromatin condensation, phosphatidylserine (PS) externalization, and the expression of an 18-kDa variant of Bax, all indicators of apoptotic cell death, and the latter suggesting the involvement of a mitochondrial route. However, procaspases-3, -8, and -9, and PARP were not cleaved, although small increases in caspase activity could be detected. At a high concentration of cisplatin, both FA and corrected cells showed a robust cleavage of procaspases and PARP. DNA fragmentation was clearly visible under high cisplatin conditions and to some extent at a low concentration in FA-A cells, but not in the FA-C cell line regardless of the presence of functional FANCC, suggesting an unknown deficiency in these cells. We conclude that hypersensitivity in FA cells is associated with a mixture of necrotic and apoptotic features that is best described as apoptotic-like cell death, and that a defective FA pathway does not interfere with the proper activation of caspase mediated cell death. PMID- 12749866 TI - Characterization of C2-ceramide-resistant HL-60 subline (HL-CR): involvement of PKC delta in C2-ceramide resistance. AB - We have established a C2-ceramide-resistant HL-60 subline (HL-CR). HL-CR cells were resistant not only to C2-ceramide but also to various anticancer drugs. HL CR cells did not respond to differentiation-inducing reagents including 1alpha,25 dihydroxyvitamin D(3), retinoic acid, and 12-O-tetradecanoylphorbol-13-acetate (TPA). TPA induced apoptosis in HL-CR cells much slower than in parental HL-60 cells. As it was reported that PKC isozymes were involved in C2-ceramide-induced apoptosis, we investigated the role of PKC isozymes in C2-ceramide resistance in HL-CR cells. The protein level of PKC delta was lower in HL-CR cells than in parental HL-60 cells, whereas the levels of PKC alpha, betaI, epsilon, and zeta were rather higher in HL-CR cells than in parental cells. Translocation of PKC delta from membrane to cytosol was induced by C2-ceramide in HL-CR cells as well as in wild-type HL-60 cells. Furthermore, overexpression of PKC delta in HL-CR cells potentiated C2-ceramide- and TPA-induced apoptosis and growth inhibition. These results suggest a role for ceramide in apoptosis and differentiation in HL 60 cells, and also suggest that PKC delta might be involved in ceramide- and TPA induced apoptosis. PMID- 12749867 TI - Great expectations of small packages. PMID- 12749868 TI - Post-extrasystolic potentiation: a viable viability test. PMID- 12749869 TI - The feasibility of ultrasonic regional strain and strain rate imaging in quantifying dobutamine stress echocardiography. AB - BACKGROUND: Ultrasonic strain rate and strain can characterize regional one dimensional myocardial deformation at rest. In theory, these deformation indices could be used to quantify normal or abnormal regional function during a dobutamine stress echo test. AIMS: The aims of our pilot study were threefold: (1) to determine the percentage of segments in which interpretable strain rate/strain data could be obtained during routine dobutamine stress echo, (2) to establish whether either the increase in heart rate or artefacts induced by respiration during dobutamine stress echo would influence analysis by degrading the data and (3) to determine the optimal frame rate vs image sector angle settings for data acquisition. Furthermore, although the detection of ischaemia was not to be addressed specifically in this study, we would describe the findings on the potential clinical role of regional deformation vs velocity imaging in detecting ischaemia-induced changes. METHODS: A standard dobutamine stress echo protocol was performed in 20 consecutive patients with a history of chest pain (16 with angiographic coronary artery disease and four with normal coronary angiograms). DMI velocities were acquired at baseline, low dose, peak dose, and recovery. To evaluate radial function (basal segment of the left ventricle posterior wall segment), parasternal LAX, SAX views were used. For long axis function data were acquired (4-CH, 2-CH views) from the septum; lateral, inferior and anterior left ventricle walls. Data was acquired using both 15 degrees (>150 frames per second (fps) and 45 degrees (115fps) sector angles. During post-processing each wall was divided into three segments: basal, mid and apical. Strain rate/strain values were averaged over three consecutive heart cycles. RESULTS: Data was obtained from 1936 segments, of which only 54 had to be excluded from subsequent analysis (2.8%) because of suboptimal quality. An increase in heart rates (up to 150/min) was not associated with a significant reduction in the number of interpretable segments. There was a significant correlation between maximal systolic strain rate/strain values obtained at narrow and at wide sector angles (e.g. a correlation for the septal segments: r=0.73,P <0.001 for strain rate, and r=0.71; P<0.001 for strain). The correlation for the timing of events obtained from narrow and wide sector angles was weaker. This would indicate that there was the insufficient temporal resolution for the latter acquisition method. Normal and abnormal regional strain rate/strain responses to an incremental dobutamine infusion were defined. In normal segments, maximal systolic strain rate values increased continuously from baseline, reaching the highest values at the peak dose of dobutamine. The segmental strain response was different. For strain, there was an initial slight increase at low dose of dobutamine (5, 10 microg/kg/min), but no further increase with increasing dose. A pattern representing an ischaemic response was identified and described. CONCLUSIONS: The feasibility study would suggest that with appropriate data collection and post-processing methodologies, strain rate/strain imaging can be applied to the quantification of dobutamine stress echo. However, appropriate post-processing algorithms must be introduced to reduce data analysis time in order to make this a practical clinical technique. PMID- 12749870 TI - Changes in regional left atrial function with aging: evaluation by Doppler tissue imaging. AB - AIMS: This study applies pulsed wave Doppler tissue imaging and colour Doppler tissue imaging to study changes in atrial function with ageing. We tested the following hypotheses: (1) pulsed wave Doppler tissue imaging can detect global changes of left atrial function associated with ageing similar to standard echocardiographic methods, (2) colour Doppler tissue imaging can reproducibly detect regional changes in atrial function (wall motion) of the normal young and normal aging atrium. METHODS AND RESULT: We studied 92 healthy subjects, divided into Group B (>or=50 years) and Group A (<50 years). As a reference standard the conventional measures of atrial function were determined: peak mitral A wave velocity, A wave velocity time integral, atrial emptying fraction and atrial ejection force. Pulsed wave Doppler tissue imaging estimated atrial contraction velocity (A' velocity) in late diastolic and segmental atrial contraction was determined by colour Doppler tissue imaging. A' velocities were significantly higher in Group B vs Group A (9.8+/-1.8 vs 8.5+/-1.5cm/s; P=0.0005). A' velocity correlated with atrial fraction (r=0.28; P=0.007) and atrial ejection force (r=0.21; P=0.04). Age correlated significantly with atrial ejection force (r=0.47; P=0.0001), atrial fraction (r=0.61; P=0.0001) and A' velocity (r=0.4; P=0.0002). Longitudinal segmental atrial contraction using colour Doppler tissue imaging showed an annular to superior segment decremental gradient with contraction velocities higher in Group B vs Group A. CONCLUSION: Pulsed wave Doppler tissue imaging and colour Doppler tissue imaging are reproducible and readily obtained parameters that provide unique data about global and segmental atrial contraction. In this study, changes in atrial contraction with aging were consistent with increased atrial contribution to filling accomplished by augmented atrial contractility. PMID- 12749871 TI - Characterizing abnormal regional longitudinal function in arrhythmogenic right ventricular dysplasia. The potential clinical role of ultrasonic myocardial deformation imaging. PMID- 12749872 TI - New insights into septal anterior wall motion velocities at end-systole in normal and hypertrophied left ventricles. AB - AIMS: It was two-fold (1) to define tissue Doppler echocardiographic characteristics of the end-systolic septal anterior motion: passive due to heart translation, or active motion free of translational effects, substantiated by a myocardial velocity gradient. (2) to specify the temporal features of this septal anterior motion on normal and hypertrophied left ventricles since it occurs while the posterior wall contracts during late ejection. METHODS AND RESULTS: Myocardial velocity gradient was calculated during the anterior motion in simultaneously colour M-mode imaged septal and posterior walls of 21 controls (49+/-12 years) and 17 patients (49+/-13 years) with left ventricle hypertrophy. Timings of septal motion were compared with flow and posterior wall motion. In controls, septal anterior motion started prior to, and overlapped the end of subaortic flow and that of the posterior wall anterior motion. Myocardial velocity gradient was found, exceeding that at the posterior wall (2.5+/-1.6 vs 0.9+/-0.5s(-1), P=0.001). In patients, septal myocardial velocity gradient was lower than in controls (1.2+/-1.04 s(-1)P=0.006). The anterior motion had a longer duration than in controls (75+/-37 vs 50+/-17ms, P=0.003). Myocardial velocity gradient and duration were correlated with septal thickness (P=<0.01). CONCLUSIONS: The septal anterior motion was active. Patients showed a decreased myocardial velocity gradient, while wall asynchrony increased. Unusual higher septal than posterior wall systolic velocities at tissue Doppler echocardiography may suggest a relaxation pattern, in spite of its end-systolic onset. PMID- 12749873 TI - Portable echocardiography: an innovative tool in screening for cardiac abnormalities in the community. AB - AIMS: Heart failure is placing an increasing burden on society. This has led to calls for echocardiographic-based programmes to screen for left ventricular systolic dysfunction and other cardiac abnormalities. Echocardiography using new fully portable echocardiography devices would allow community-based cost effective screening programmes once validated. This study was undertaken to evaluate this further in both high and low-risk subjects. METHODS AND RESULTS: 562 consecutive subjects attending a community-based heart failure screening programme, some at high-risk and some at low-risk of cardiac abnormalities, underwent echocardiography by both portable and traditional echocardiography machines. An 'eyeball' estimate of left ventricular ejection fraction was made on the portable device and compared to a quantitative measure of ejection fraction on the traditional machine. Qualitative measures of valvular regurgitation and quantitative measures of left ventricular hypertrophy were also compared. An estimate of ejection fraction was possible in 97% of cases using portable echocardiography. It gave a sensitivity, specificity and negative predictive value in diagnosing left ventricular systolic dysfunction of 96%, 98% and 99.6%, respectively. Inter-observer variability gave a mean difference in ejection fraction of 2%, and 95% limits of agreement of -8% to +12%. All cases of moderate or severe valvular regurgitation and 29 of 31 cases of significant left ventricular hypertrophy were correctly identified as abnormal on the portable device. CONCLUSIONS: Thus, echocardiography performed by experienced sonographers using these new fully portable devices is an accurate and reproducible technique for detecting left ventricular systolic dysfunction, left ventricular hypertrophy and valvular regurgitation in both high-risk and low-risk members of the community. Its very high negative predictive values would allow their use in future community-based screening programmes. PMID- 12749875 TI - Spontaneous recovery of myocardial asynergic segments following acute myocardial infarction. The role of post-extrasystolic potentiation echocardiography in the predischarge evaluation. AB - AIMS: Predischarge identification of viable myocardium with a spontaneous recovery over time can be helpful in decision-making process following acute myocardial infarction (AMI). An intriguing hypothesis is that identification of viability in myocardial asynergic segments with or without spontaneous recovery over time may require an inotropic stimulus of a different intensity. To test this we used post-extrasystolic potentiation (PESP), which represents an inotropic stimulation of a different intensity as a function of coupling interval. METHODS AND RESULTS: Myocardial viability was assessed by PESP echocardiography in 86 patients with a first uncomplicated AMI. Spontenous changes in contractile function of viable but asynergic segments at predischarge were evaluated by a follow-up resting echocardiogram 1 month later. Viable myocardium of left ventricular asxynergic segments with spontaneous recovery or persistence of dysfunction showed its first significant improvement for different values of coupling interval during PESP (409+/-18 vs 336+/-23ms (milliseconds), P<0.0001). An arbitrary cut-off value for RR interval >or=380ms, correctly identified 91% of myocardial segments undergoing spontaneous recovery, while 99% of asynergic myocardial segments with spontaneous recovery had a threshold coupling interval >or=380ms. CONCLUSION: At predischarge following AMI, asynergic but viable myocardial segments by PESP with spontaneous recovery of contraction over time require an inotropic stimulus of lesser intensity to acutely improve contraction pattern than viable and asynergic segments without this favourable outcome. PMID- 12749874 TI - Myocardial performance index as predictor of adverse outcomes following mitral valve surgery. AB - AIMS: We aim to determine whether the myocardial performance index, will be a good predictor of adverse outcomes following mitral valve surgery. METHOD: We prospectively measured pre-operative myocardial performance index in 22 consecutive patients, with moderate to severe mitral insufficiency, undergoing corrective mitral valve surgery. The primary endpoint was predefined as either peri-operative death or congestive heart failure. RESULTS: The primary endpoint occurred in nine patients. Five of the six patients with myocardial performance index >or=0.7 had primary endpoints. Chi-square testing demonstrated that the primary endpoint was significantly associated with advanced age (>or=70 years) and myocardial performance index >or=0.7 (P=0.003 and 0.01 respectively). There was a trend towards significant association of depressed left ventricle ejection fraction (left ventricle ejection fraction or=0.7. CONCLUSION: Our results suggest that myocardial performance index is a potentially useful predictor of increased risk of peri-operative death or congestive heart failure, in patients with moderate-severe mitral insufficiency undergoing corrective mitral valve surgery. In conjunction with left ventricle ejection fraction, it may be helpful in the pre-operative prognostication of these patients. PMID- 12749876 TI - The use of small personal ultrasound devices by internists without formal training in echocardiography. AB - AIMS: Hand-held ultrasound devices will probably be used for bedside cardiac diagnoses by internists without formal training in echocardiography. We compared the accuracy of hand-held ultrasound devices studies performed by expert echocardiographers vs internal medicine residents with brief training in echocardiography. METHODS AND RESULTS: Three internal medicine residents participated in an organized training program in echocardiographic principles, image acquisition, and interpretation. Subsequently, these residents and three echocardiographers imaged 300 patients with a hand-held ultrasound device. Sensitivity, specificity, positive and negative predictive values for internist- and echocardiographer-performed studies for the detection of cardiac abnormalities were compared using a full-featured exam as the gold standard. Resident- and echocardiographer-performed scans had similar overall sensitivity and specificity. There was a higher positive predictive value for the echocardiographer-performed scans. For clinically important findings (likely to affect patient care), sensitivity was slightly but significantly higher for the echocardiographer-performed scans. Clinically important findings most often missed by residents included regional wall motion abnormalities, intra-cardiac thrombus, right ventricular dysfunction and non-trivial pericardial effusions. CONCLUSION: Hand-held ultrasound devices provide useful screening tools for cardiac disease but should not replace a standard platform study. Training guidelines and competency evaluation are needed if these devices are to be used by non-echocardiographers for clinical decision-making. PMID- 12749878 TI - Intramyocardial dissection with rupture of the right ventricle after an acute myocardial infarction--a rare complication diagnosed by transesophageal echocardiography. PMID- 12749877 TI - Post-extrasystolic potentiation recruits incremental contractile reserve of dyssynergic myocardium during dobutamine stress testing: evidence by pulsed wave tissue Doppler imaging. AB - Dobutamine stress echocardiography is an established diagnostic method for the detection of myocardial viability in patients with severe left ventricular dysfunction([1]). The presence of viable myocardium identifies patients who will benefit from coronary revascularization, by improving both functional capacity and long-term survival. Occasionally, dobutamine infusion has been combined with other stressors, such as post-extrasystolic potentiation, in order to improve accuracy. The contractile reserve after combined dobutamine infusion and post extrasystolic potentiation can be quantified by pulsed wave tissue Doppler imaging. We describe a patient with severe left ventricular dysfunction, in which pulsed wave tissue Doppler imaging allowed to demonstrate that post-extrasystolic potentiation superimposed on dobutamine infusion is able to further recruit contractile reserve, as compared to dobutamine infusion alone. A nuclear scan assessing glucose utilization was used as a reference. PMID- 12749879 TI - Giant multiperforated atrial septal aneurysm in a patient with paroxysmal atrial fibrillation. AB - With the widespread use of transoesophageal echocardiography, atrial septal aneurysm (ASA) is frequently and easily recognized. Diagnostic criteria, however, remain arbitrary. A meticulous search for interatrial shunting (most often patent foramen ovale (PFO)) and associated cardiac diseases is important. ASA is considered a risk factor for cardioembolism, especially in association with patent foramen ovale. We describe a patient with a giant multiperforated ASA presenting with refractory paroxysmal atrial fibrillation. Resection of the aneurysm was performed in combination with a surgical Maze procedure. PMID- 12749881 TI - Inhibition of HIV tat-TAR interactions by an antisense oligo-2'-O methylribonucleoside methylphosphonate. AB - An antisense oligo-2'-O-methylribonucleotide having alternating methylphosphonate/phosphodiester linkages, 1676, whose sequence is complementary to the apical stem-loop of HIV-1 TAR RNA, was prepared to determine its effects on Tat protein-TAR interaction and Tat-mediated gene transactivation in cell culture. This oligomer and its all-phosphodiester analogue, 1707, were shown to: (1) bind to TAR at 37 degrees C with K(d)'s in the low nM concentration range; (2) inhibit Tat-TAR complex formation; and (3) inhibit expression of a chloramphenicol reporter gene under control of the HIV LTR in HeLa HL3T1 cells in culture. PMID- 12749882 TI - Synthesis and biological evaluation of some novel 4'-thio-L-ribonucleosides with modified nucleobase moieties. AB - 1,2,3,5-tetra-O-acetyl-4-thio-beta-L-ribofuranose (13) was synthesized by an improved five-step sequence starting from methyl alpha-D-lyxopyranoside. Compound 13 was then converted to the corresponding L-4'-thionucleosides 4-6 and 19 by a modified Vorbruggen procedure. All of these nucleoside analogues were tested for their antitumour activity in vitro. PMID- 12749883 TI - Synthesis and biological evaluation of new UDP-GalNAc analogues for the study of polypeptide-alpha-GalNAc-transferases. AB - A series of three O-methylated UDP-GalNAc analogues have been synthesised using a divergent strategy from a 3,6-di-O-pivaloyl GlcNAc derivative. The biological activity of these probes toward polypeptide-alpha-GalNAc-transferase T1 has been investigated. This study shows that this glycosyltransferase exhibits a very high substrate specificity. PMID- 12749884 TI - Acyclic N-(azacycloalkyl)bisindolylmaleimides: isozyme selective inhibitors of PKCbeta. AB - The synthesis and structure-activity relationship (SAR) trends of a new class of N-(azacycloalkyl)bisindolylmaleimides 1, acyclic derivatives of staurosporine, is described. The representative compound for this series (1e) exhibits an IC(50) of 40-50 nM against the human PKCbeta(1) and PKCbeta(2) isozymes and selectively inhibits the PKCbeta isozymes in comparison to other PKC isozymes (alpha, gamma, delta, epsilon, lambda, and eta). The series is also kinase selective for PKC in comparison to other ATP-dependent kinases. A comparison of the PKC isozyme and kinase activity of the series is made to the kinase inhibitor staurosporine. PMID- 12749885 TI - Amide analogues of TSA: synthesis, binding mode analysis and HDAC inhibition. AB - The synthesis of new amide type histone deacetylase inhibitors is described, having an (R)-methyl substituent and a diene or saturated structure of the chain linking the hydroxamic acid and dimethylaminobenzoyl groups. The saturated compound shows stronger HDAC inhibition than the unsaturated analogue. Molecular modeling suggests that the flexibility of the linker chain is important for an optimal orientation of the dimethylaminobenzoyl group in the enzyme. PMID- 12749886 TI - Binding mode of 6ECDCA, a potent bile acid agonist of the farnesoid X receptor (FXR). AB - Based on the folding conservation across the nuclear receptor superfamily and the sequence homology with RAR-gamma, we report the construction of a three dimensional model of the ligand binding domain of FXR. The model is exploited for the elucidation of the binding mode of 6alpha-ethyl-chenodeoxycholic acid. The results of the docking experiments give quite clear indications that the bile acid derivative would bind the receptor in a mode significantly different than that observed for agonists of other nuclear receptor superfamily. PMID- 12749887 TI - Density functional theory calculations for resveratrol. AB - The calculations based on the density functional theory (DFT) have been used to study the structure-activity of resveratrol in the chain reaction of autooxidation. According to the geometry obtained by using a B3LYP/6-31G**, the HOMO, LUMO of resveratrol and the spin density, the single electron distribution of the 4'- and 5-radical of resveratrol were calculated, it was found that resveratrol is a potential antioxidant. The 4'-hydroxyl group of resveratrol is more reactive than 3- and 5-positions because of the resonance effects. The dominant structure of the resveratrol radicals is a semiquinone structure which determines the stability of radicals, and the unpaired electron is mainly distributed to the O-atom and its ortho and para positions. The antioxidant activity of resveratrol is related to the spin density and the unpaired electron distribution of the O-atom. PMID- 12749888 TI - Synthesis, pharmacology and pharmacokinetics of 3-(4-aryl-piperazin-1-ylalkyl) uracils as uroselective alpha1A-antagonists. AB - Predominance in the urethra and prostate of the alpha(1A)-adrenoceptor subtype, which is believed to be the receptor mediating noradrenaline induced smooth muscle contraction in these tissues, led to the preparation of alpha(1A) selective antagonists to be tested as uroselective compounds for the treatment of benign prostatic hyperplasia. Thus, a number of selective alpha(1A)-adrenoceptor antagonists were synthesized and assayed in vitro for potency and selectivity. Dog pharmacokinetic parameters of 12 (RO700004) and its metabolite 40 (RO1104253) were established. The relative selectivity of intravenously administered 12, 40 and standard prazosin to inhibit hypogastric nerve stimulation-induced increases in intraurethral prostatic pressure versus phenylephrine-induced increases in diastolic blood pressure in anesthetized dogs was 76, 71 and 0.6, respectively. PMID- 12749889 TI - Design and synthesis of novel pyrrolidine-containing bradykinin antagonists. AB - The design and synthesis of novel pyrrolidine-containing bradykinin antagonists, II, are described. Conformational analysis suggested that a pyrrolidine moiety could substitute for the N-methyl cis-amide moiety of FR 173657. The in vitro binding data showed that the (S)-isomer of II was potent in the bradykinin B(2) receptor-binding assay with a K(i) of 33 nM. The opposite isomer, (R)-II, had a K(i) of 46 nM. The in vitro binding data confirmed our conformational hypothesis. PMID- 12749890 TI - Optimization of alpha-acylaminoketone ecdysone agonists for control of gene expression. AB - Fifteen new alpha-acylaminoketones were prepared by four different routes in an initial effort to optimize the potency of these compounds as ecdysone agonists. The compounds were assayed in mammalian cells expressing the ecdysone receptors from Bombyx mori (BmEcR) and Choristoneura fumiferana (CfEcR) for their ability to cause expression of a reporter gene downstream of an ecdysone response element. A new alpha-acylaminoketone was identified which had activity equal to that of the standard dibenzoylhydrazine ecdysone agonist GS()-E in the assay based on CfEcR. PMID- 12749892 TI - N-isonicotinoyl-(L)-4-aminophenylalanine derivatives as tight binding VLA-4 antagonists. AB - A series of isonicotinoyl-(L)-aminophenylalanine derivatives was prepared and evaluated as VLA-4 antagonists. These compounds exhibit subnanomolar binding affinity to VLA-4 and significant off-rates. The interplay between off-rate, protein binding and pharmacokinetics is discussed. PMID- 12749893 TI - Indole amide hydroxamic acids as potent inhibitors of histone deacetylases. AB - A series of hydroxamic acid-based HDAC inhibitors with an indole amide residue at the terminus have been synthesized and evaluated. Compounds with a 2-indole amide moiety have been found as the most active inhibitors among the different regioisomers. Introduction of substituents on the indole ring further improved the potency and generated a series of very potent inhibitors with significant antiproliferative activity. A representative compound in the series, 7b, has been found to be orally active in tumor growth inhibition model. PMID- 12749894 TI - Advances toward new antidepressants beyond SSRIs: 1-aryloxy-3-piperidinylpropan-2 ols with dual 5-HT1A receptor antagonism/SSRI activities. Part 1. AB - A series of 1-aryloxy-3-piperidinylpropan-2-ols possessing potent dual 5-HT(1A) receptor antagonism and serotonin reuptake inhibition was discovered. 1-(1H-Indol 4-yloxy)-3-(4-benzo[b]thiophen-2-ylpiperidinyl)propan-2-ols exhibited selective and high affinity at the 5-HT(1A) receptor and serotonin reuptake inhibition at nanomolar concentrations for dual activities. PMID- 12749891 TI - Potent, selective inhibitors of protein tyrosine phosphatase 1B. AB - We have previously reported a novel series of oxalyl-aryl-amino benzoic acid based, catalytic site-directed, competitive, reversible protein tyrosine phosphatase 1B (PTP1B) inhibitors. With readily access to key intermediates, we utilized a solution phase parallel synthesis approach and rapidly identified a highly potent PTP1B inhibitor (19, K(i)=76 nM) with moderate selectivity (5-fold) over T-cell PTPase (TCPTP) through interacting with a second phosphotyrosine binding site (site 2) in the close proximity to the catalytic site. PMID- 12749895 TI - Tetrahydroquinoline-based selective estrogen receptor modulators (SERMs). AB - A new series of estrogen receptor ligands based on a 6-hydroxy tetrahydroquinoline scaffold is described, in addition to their binding affinity and functional activity in MCF-7 cells. Several 1,2-disubstituted tetrahydroquinolines bearing a basic side chain were shown to be high affinity ligands and antagonists in the MCF-7 proliferation assay. Compounds lacking the basic side chain were agonists in the MCF-7 assay. PMID- 12749896 TI - Redefining the structure-activity relationships of 2,6-methano-3-benzazocines. Part 2: 8-formamidocyclazocine analogues. AB - High affinity binding for mu and kappa opioid receptors has been observed in analogues of cyclazocine, ethylketocyclazocine and naltrexone where the prototypic (of opiates) phenolic OH group was replaced with a formamide (-NHCHO) group. For the 8-formamide analogue of cyclazocine, binding is highly enantiospecific (eudismic ratios approximately 2000 for mu and kappa) with K(i) values 0.96). The catheter position within the flow phantom and the size of the ring-down were theoretically analyzed. These elements, and noise in the RF signals, have an important influence on the IVUS blood flow measurements reflected by the offset and the slope of the linear relationships. By placing the IVUS catheter outside the flow phantom, parabolic blood flow profiles were also measured. Second, IVUS blood flow measurements were performed in the carotid artery of two Yorkshire pigs, which showed linear relationships (all R(2)> 0.85) between the IVUS-derived flow and the calibrated EMF. Experimentally, the offset was lower than 3 mL/min and the slope was close to 1. Third, IVUS blood flow measurements were performed in coronary arteries in patients. Preliminary results for the coronary flow reserve (CFR = high flow/baseline flow) in patients using the decorrelation method of RF signals of an array IVUS scanner were comparable with CFR based on Doppler measurements. PMID- 12749922 TI - Implementation issues in ultrasonic flow imaging. AB - This article addresses several implementation issues in ultrasonic flow imaging. We discuss frequency-dependent scattering and attenuation, use of interpolation for computation intensive methods and implications of the use of chirps to increase bandwidth. We also discuss wall filtering issues; our observations show that the butterfly search estimator may be capable of detecting flow in the vicinity of strong stationary scatterers (clutter) without additional processing such as wall-filtering. Illustrative examples are given for simulated and experimental data. PMID- 12749923 TI - High-definition freehand 3-D ultrasound. AB - This paper describes a high-definition freehand 3-D ultrasound (US) system, with accuracy surpassing that of previously documented systems. 3-D point location accuracy within a US data set can be achieved to within 0.5 mm. Such accuracy is possible through a series of novel system-design and calibration techniques. The accuracy is quantified using a purpose-built tissue-mimicking phantom, designed to create realistic clinical conditions without compromising the accuracy of the measurement procedure. The paper includes a thorough discussion of the various ways of measuring system accuracy and their relative merits; and compares, in this context, all recently documented freehand 3-D US systems. PMID- 12749924 TI - Contrast superharmonic imaging: a feasibility study. AB - Harmonic imaging provided significant improvement in image quality by taking advantage of the scattered second harmonic (2H) component from contrast bubbles. However, differentiation between contrast and tissue (usually termed contrast-to tissue ratio, CTR) is sometimes cumbersome and this is mainly due to tissue contamination. We have previously demonstrated, using simulations and in vitro measurements, that CTR increases as a function of the order of the harmonic number. A new contrast imaging method based on the detection of the higher harmonics was developed and termed superharmonic (SH). This technique has been shown to be more sensitive to contrast by increasing the signal from contrast and suppressing that from tissue (high CTR). The purpose of this study was to determine the clinical feasibility and usefulness of SH in patients using a commercially available contrast agent (SonoVue(R)) for quantification of myocardial perfusion. A total of 10 patients with various cardiac diseases were assessed. Apical four-chamber views were acquired using SH in triggered mode before and after contrast injection. The superharmonic was performed with a newly developed probe transmitting at 0.8 MHz with a mechanical index of 0.2. Myocardial perfusion was determined visually and analyzed quantitatively using radiofrequency (RF) processing from different regions of interest. The results showed that, before contrast injection, SH was totally blinded to tissue and no superharmonic components were generated in the image view. After administration of SonoVue(R), myocardial opacification was visualized by SH after contrast entered the myocardium. An increase of more than 15 dB in the myocardial bubbles echo compared to tissue echo was measured. In addition, the technique was used to visualize myocardial perfusion after myocardial septal ablation for hypertrophic cardiomyopathy. The clinical results showed the ability of contrast SH imaging in differentiating low and normal perfusion areas, demonstrating the high sensitivity and specificity of the technique. PMID- 12749925 TI - Nonlinear behaviors of contrast agents relevant to diagnostic and therapeutic applications. AB - The nonlinear properties of an encapsulated microbubble of a contrast agent were studied theoretically and experimentally. A modified nonlinear differential equation (Herring equation) was used to describe the radial oscillation of the microbubble and solved numerically. It was found that the nonlinear resonance frequency, at which the peak radial oscillation amplitude occurs, was a decreasing function of the acoustic amplitude of a driving ultrasonic pulse. Optical images of the contrast agent microbubbles under various ultrasonic exposure conditions: 1. sham exposure; 2. 2-MHz spatial peak acoustic pressure = 200 kPa, I(SATA) = 260 mW/cm(2), duty cycle = 7.5%, repetition period = 0.0266 ms; 3. 0.5-MHz spatial peak acoustic pressure = 200 kPa, I(SATA) = 130 mW/cm(2), duty cycle = 7.5%, repetition period = 0.1067 ms; have also shown that the lower frequency ultrasound (US) excitation (0.5 MHz) is more effective in disruption of the microbubbles due to acoustic inertial cavitation than the higher frequency US (2 MHz). PMID- 12749926 TI - On the destruction of microbubble ultrasound contrast agents. AB - In recent years, the use of microbubble ultrasound (US) contrast agents as carriers in drug and gene delivery applications has intensified the need for a clear understanding of the processes involved in their destruction. In this study, an analysis of the conditions in the shell of a contrast agent particle has been made, based on the full numerical solution of a modified Rayleigh Plesset equation. The results indicate that extremely high shell stresses may be expected under typical clinical conditions. Examination of previous experimental evidence in the light of these findings suggests that the shells are almost invariably disrupted, even if they are not visibly destroyed. This has some serious implications, both for targeted delivery processes and reliable assessment of the potential for harmful bioeffects. At present, neither the model nor the experimental data provide an adequate description of contrast agent behaviour. This is due primarily to the lack of information regarding the mechanical response of the shell material and the restriction of the model to the case of small, spherically symmetrical oscillations. Methods for addressing these deficiencies in future work are proposed. PMID- 12749928 TI - Numerical simulation of the transient temperature field from an annular focused ultrasonic transducer. AB - Knowledge of the extent of the "heated necrosis element" from a single exposure in target tissue created by an ultrasonic beam is critical for the application of focal ultrasound (US) surgery (FUS). This study uses the O'Nell and Pennes formulas to simulate the heated necrosis element from an annular focused transducer and to examine its dependence on exposure dosage, as well as some design parameters of the transducer. Several conclusions may be drawn from our numerical results: 1. With increasing exposure, the heated necrosis element increases, but its contour becomes plumper and the influence of sound intensity I is found to be greater than that of the exposure time t. 2. To get a similar heated necrosis element, the exposure approximately satisfies a relation: It(0. 4 3)=constant. 3. Increasing the US frequency or the outer-radius of the annular transducer leads to a decrease in the heated necrosis volume. PMID- 12749927 TI - Angle-independent estimation of maximum velocity through stenoses using vector Doppler ultrasound. AB - Categorisation for arterial stenoses treatment is determined primarily by the degree of occlusion, which is often estimated ultrasonically from blood velocity measurements. In current single-beam ultrasound (US) systems, this estimate can suffer from gross errors due to angle-dependence. The purpose of this study was to find out if an experimental dual-beam US system could reduce the angle dependence of the velocity estimates. We compared four dual-beam velocity estimation algorithms on both a string phantom and straight tube wall-less flow phantoms incorporating symmetrical and asymmetrical stenoses from 0% to 91% by area. The estimated maximum velocity varied, on average, by 7.6% for beam-vessel angles from 40 degrees to 80 degrees. The fluctuation in the magnitude estimate was reduced by a factor of 2.6 using a hybrid single-dual-beam algorithm. We conclude that, when the true velocity lies in the scan plane, the dual-beam system reduces the angle-dependence and, thus, has the potential to improve categorisation of patients with arterial stenoses. PMID- 12749929 TI - Effect of split-focus approach on producing larger coagulation in swine liver. AB - The split-focus approach has the potential to substantially improve the throughput of coagulation high-intensity focused ultrasound (HIFU) treatment. A prototype split-focus transducer with four elements at 4.25 MHz, combined with a small imaging probe at 6.5 MHz, was constructed for transrectal treatment of a prostate. Computer simulation predicted that 1.57 times larger acoustic power than a single-spot focus was required for a split focus to heat the tissue up to the same peak temperature at 4 s from the start of insonation. When the peak temperature was 100 degrees C, the tissue volume above 70 degrees C was predicted to be 2.3 times larger for a split focus. Swine liver lobes were intraoperatively insonated with the split-focus transducer for 4 s. A lesion of coagulation necrosis 3 to 4 times larger in volume than with a single-spot focus was formed with a split focus at the same acoustic power ratio as described above. Furthermore, a lesion about 2 times larger in volume was formed when compared at the same acoustic power. These results show the great advantage of a split-focus approach over a conventional single-spot focus in coagulation treatment. PMID- 12749931 TI - Molecular damage to membrane proteins induced by ultrasound. AB - A dilute suspension of human erythrocytes was exposed in vitro to ultrasound (US) with a frequency of 1 MHz and an acoustic pressure of (3.0 +/- 0.4) x 10(5) Pa (intensity of 2.9 +/- 0.8 W/cm(2)). The degree of lysis of the cells in saline increased with increasing the exposure time. An addition of a small amount of plasma to the suspension did not change the degree of lysis. A gel electrophoresis of membrane proteins extracted from the exposed and lysed membrane samples showed that the exposure caused molecular damage in membrane proteins in the absence of plasma, and that the addition of the small amount of plasma protected the proteins against the molecular damage. PMID- 12749930 TI - DNA transfer and cell killing in epidermoid cells by diagnostic ultrasound activation of contrast agent gas bodies in vitro. AB - DNA transfer by sonoporation and cell killing in monolayer cells were examined by contrast-aided low-power diagnostic ultrasound (US). Culture chambers with epidermoid cell monolayers were scanned at about 1 mm/s with a 1.5-MHz scan head aimed upward at the chamber in a 37 degrees C water bath. For DNA transfer tests, plasmids coding for green fluorescent protein (GFP) were added to the medium, and GFP expression was assessed by flow cytometry after 2 days. In separate tests, cell killing was determined immediately after treatment. GFP-positive cell counts were 0.4% (0.7% SD) for shams and 3.7% (1.2% SD) of cells for exposure at 2.3 MPa with 2% Optison contrast agent. The fraction of dead cells was 3.4% (1.7% SD) in shams and 28.6% (6.3% SD) in exposed chambers. Both effects increased for increasing Optison concentration and increasing peak rarefactional pressure amplitude. Contrast-aided diagnostic US has a potential therapeutic application for gene transfer, but a trade-off appears to exist with cell killing. PMID- 12749932 TI - Left ventricular myocardial function in congenital valvar aortic stenosis assessed by ultrasound tissue-velocity and strain-rate techniques. AB - A pilot study was performed to reveal the potentials of new echo Doppler techniques for the detection of myocardial changes due to congenital valvar aortic stenosis. A total of 24 patients, (age range 0.1 to 17 years), with various degrees of aortic stenosis, and 24 age- and gender-matched, healthy children were enrolled in this study. Conventional echo Doppler, tissue velocity imaging (TVI) and strain-rate imaging (StRI) measurements were carried out using the apical four-chamber view and transthoracic long-axis view. All patients had normal fractional shortening of the left ventricle (> 28%). Although the sum of septal and ventricular wall thicknesses was significantly increased in the patients (p < 0.001), only 6 of the 24 patients showed left ventricular hypertrophy. In tissue velocity mode, systolic and early diastolic wall velocity acceleration was significantly reduced in both views. Peak systolic and early diastolic wall velocities, as well as strain rate values, in the four-chamber view were significantly reduced in the patient group. The decrease was highest for the strain-rate values in all cases. In conclusion, strain rate values at different moments within the heart cycle might become important parameters in the assessment of myocardial impairment. Further studies are indicated to assess the correlation of these parameters with the severity of stenosis, left ventricular hypertrophy and irreversible myocardial function changes. PMID- 12749933 TI - A pulsating coronary vessel phantom for two- and three-dimensional intravascular ultrasound studies. AB - The evaluation of new techniques for 2-D and 3-D intravascular ultrasound (US) imaging (IVUS) often requires the use of a pulsating coronary phantom. This study describes the design, construction and evaluation of a phantom simulating the pulsation of a human coronary artery for IVUS studies. Polyvinyl alcohol (PVA) cryogel was used as a tissue mimic for the coronary vessel, which was incorporated in a custom-built assembly. The phantom was programmed to pulsate under servomotor control, to model the pulsation of a normal coronary artery and 2-D IVUS images were obtained using an IVUS imaging catheter. To evaluate the performance of the phantom, the lumen area variation of the phantom was determined and compared with the programmed pulsation waveforms. Our results showed that phantom pulsation correlated well with the programmed pulsation waveform (r = 0.97). The deviation of the least squares line from the line of identity was calculated to be < 4%. PMID- 12749934 TI - Evaluation of spermatological parameters in ochratoxin A--challenged boars. AB - Exposure to certain mycotoxins has been proved to contribute to fertility problems in pigs. Although ochratoxin A (OA) is one of the most common naturally occurring mycotoxins, there is little data concerning the possible effects of this toxin on sperm quality of boars. After a 4-week control period, animals were given 20 microg OA per os daily for 6 weeks, followed by a 9-week withdrawal period. Serum and seminal plasma were monitored for OA with enzyme-linked immunosorbent assay (ELISA). Spermatozoal motility was measured at 0, 24, 48, 72 and 96 h, and ejaculation volume, initial viability and progressive motility were recorded. Samples of testis and epidydimidis were evaluated histologically. Viability, initial forward motility, and motility after 24h storage were significantly reduced in the experimental group in the withdrawal period only. There were no major histological differences in number and morphology of Leydig cells and epydidimal structures between experimental and control boars. Results of the present study demonstrate that OA may affect sperm production and boar semen quality only after a lag period. Further research is required to elucidate the possibility of a direct or indirect interaction between the toxin and germ cells (spermatogonia). PMID- 12749936 TI - Identification, characterization and localization of three proteins expressed by the porcine oviduct. AB - At estrus, the oviduct undergoes endocrine-induced changes which provide an essential microenvironment for maturation of gametes, fertilization and embryonic development. Several oviduct expressed proteins which interact with gametes or embryos, including the oviduct-specific, estrogen-dependent glycoprotein (OGP), have been identified and characterized. The objective of the present study was to identify, characterize and localize other proteins expressed by the porcine oviduct during estrus that may function in an autocrine or paracrine manner to enhance fertilization and embryonic development. Oviducts were collected during the estrous cycle or early pregnancy, flushed and divided into functional segments, and portions of the infundibulum, ampulla and isthmus were fixed for immunocytochemical analysis or cultured. Culture media was semi-purified by heparin-agarose affinity chromatography, proteins were transferred to polyvinylidene fluoride (PVDF) membrane after two-dimensional (2D)-SDS-PAGE and three different proteins were identified, excised and subjected to N-terminal amino acid analysis. These proteins were identified as complement component C3b, the carboxy-terminal propeptide of alpha 1 (III) procollagen (PIIICP), and the heavy chain variable region of IgA. Electrophoresis and fluorography of media from Days 0 to 12 of early pregnancy or the estrous cycle revealed both spatial and temporal expression of C3b and IgA heavy chain but not PIIICP by the oviduct. Further, all three proteins were identified in oviduct fluid by electrophoresis, immunoblot or immunoprecipitation analysis. Complement component C3b and IgA heavy chain were immunolocalized in all three oviduct segments on all days; however, temporal and spatial differences were demonstrated. Staining was greater in the infundibulum and during estrus for all three identified proteins. In summary, three proteins expressed by the oviduct at estrus and during early pregnancy were identified; characterization and localization suggest they may play a critical role in protecting the luminal environment, participating in ECM remodeling and gamete interactions. PMID- 12749935 TI - Development and comparison of in vivo and in vitro models for endometritis in cows and mares. AB - In order to investigate pathogenic mechanisms of acute endometritis in cows and mares, we established an in vivo model in both species. Based on the results of an in vitro transmigration system, human recombinant interleukin-8 (rhIL-8; 1.25 microg per mare and 5 microg per cow in 50 ml phosphate-buffered saline) was used to attract polymorphonuclear neutrophil granulocytes (PMNs) into the uteri. Peak numbers of uterine neutrophils were attracted after 6h, in both cows and mares. On average, mares responded more sensitively than cows, with 15 times higher numbers of rhIL-8-attracted uterine neutrophils (72+/-8 x 10(7)cells). In contrast to in vitro studies, in vivo migrated neutrophils (uterine neutrophils) of both species displayed a significantly reduced MHC class I expression. Expression of the CD11a molecule was significantly enhanced on equine uterine neutrophils but downregulated on bovine cells. Compared with untreated autologous peripheral neutrophils, both uterine and in vitro migrated neutrophils showed no alteration of phagocytic capacity. The ability to generate reactive oxygen species (ROS) was significantly upregulated in bovine and equine uterine neutrophils. This was also observed after in vitro migration of equine neutrophils, whereas ROS generation by bovine neutrophils was significantly depressed. In summary, the concept of inducing endometritis directly by local application of human interleukin-8 has been reliably successful in cows and mares. The model permits the analysis of PMN migration into the uterus under defined and controlled conditions. The observed differences between cows and mares with respect to phenotypical and functional characteristics of in vivo attracted uterine cells point to species-related features of neutrophil migration. In vitro transmigrated bovine and equine cells partially differ in phenotype and function from uterine neutrophils. Therefore, the in vitro transmigration assay cannot completely represent the in vivo endometritis model described here. PMID- 12749937 TI - Development of in vitro matured, in vitro fertilized domestic cat embryos following cryopreservation, culture and transfer. AB - The ability of embryos to successfully survive cryopreservation is dependent on both morphological and developmental characteristics. Domestic cat oocytes matured in vitro exhibit alterations in nuclear and cytoplasmic maturation that may affect developmental competence, particularly after cryopreservation. In Experiment 1, we evaluated the developmental competence of in vitro produced (IVM/IVF) cat embryos after cryopreservation on Days 2, 4 or 5 of IVC. In Experiment 2, in vivo viability was examined by transfer of cryopreserved embryos into recipient queens. Oocytes recovered from minced ovaries were cultured in TCM 199 with hCG/eCG and EGF at 38 degrees C in 5% O(2), 5% CO(2), 90% N(2) for 24h. In Experiment 1, after IVM/IVF, on Day 2 (n=56), Day 4 (n=48) and Day 5 (n=42) of IVC, embryos were equilibrated for 10 min at 22 degrees C in HEPES (15m M) Tyrode's (HeTy) with 1.4M propylene glycol (PG), 0.125 M sucrose (S), 10% dextran and 10% FBS, loaded into 0.25 ml straws, cooled at 2.0 degrees C/min to -6.0 degrees C and held for 10 min. After seeding, cooling resumed at 0.3 degrees C/min to -30 degrees C and after a 10 min hold, straws were plunged into liquid nitrogen (LN(2)). Straws were thawed in air for 2 min and cryoprotectant was removed by a five-step rinse consisting of 3 min each in HeTY with 0.95 M PG/0.25 M S; 0.95 M PG/0.125 M S; 0.45 M PG/0.125 M S; 0 PG/0.125 M S; 0 PG/0.0625 M S. Contemporary IVM/IVF embryos were used as nonfrozen controls (Day 2, n=14; Day 4, n=26; Day 5, n=35). After 8 days of IVC, the number of embryos developing to blastocysts was recorded and blastocyst cell numbers were counted after staining with Hoechst 33342. In Experiment 1, developmental stage did not affect the survival rate after thawing (Day 2=79%, Day 4=90%, Day 5=98%) and was not different from that of controls (Day 2=89%, Day 4=88%, Day 5=96%). Blastocyst development was similar among days both after cryopreservation (Day 2=59%, Day 4=54%, Day 5=63%) and in controls (Day 2=55%, Day 4=54%, Day 5=58%). Mean (+/ S.D.) cell number of blastocysts was slightly lower (NS) in cryopreserved embryos (Day 2=152+/-19, Day 4=124+/-20, Day 5=121+/-24) than in controls (Day 2=141+/ 25, Day 4=169+/-21, Day 5=172+/-19). In Experiment 2, embryos frozen on Day 2 (n=68), Day 4 (n=49) or Day 5 (n=73) were thawed and cultured for 3, 1, or 0 days before transfer by laparotomy to 5 (mean=12.6+/-2.4), 4 (mean=12.2+/-3.7) and 6 (mean=12.0+/-1.6) recipients, respectively. Four recipients were pregnant on Day 21; two from embryos frozen on Day 4 and two from Day 5. Two live kittens were born at 66 days, a third kitten died during parturition at 64 days and a fourth pregnancy aborted by Day 45. In summary, we have shown that a controlled rate cryopreservation technique can be successfully applied to cat embryos produced by IVM/IVF. In vitro development to the blastocyst stage was not affected by the age of embryos at cryopreservation. The births of live kittens after ET of cryopreserved embryos is additional validation of progress toward applying assisted reproductive technology to preservation of endangered felids. PMID- 12749938 TI - Successful production of piglets derived from vitrified morulae and early blastocysts using a microdroplet method. AB - This study was conducted to determine the efficiency of vitrification using the microdroplet (MD) method for early stage porcine embryos. Embryos at compacted morulae to early blastocyst stage were vitrified in a vitrification solution containing 40% (v/v) ethylene glycol, 0.6M sucrose and 2% (w/v) polyethylene glycol in M2 (ESP) without any pretreatment. The equilibration and dilution were carried out in third and fourth steps, respectively, at 38 degrees C. The survivability of the cryopreserved embryos was assessed for both in vitro culture (Experiment 1) and by embryo transfer (Experiment 2). In Experiment 1, the embryos were vitrified within a microdroplet or 0.25 ml straw (ST) and fresh embryos were used as a control group. The survival rates after 24h culture in the MD, ST and control groups were 21/23, 14/20 and 20/20, respectively. The hatching rates of the embryos after 48 h incubation were 14/23, 4/20 and 16/20, respectively. In Experiment 2, 171 vitrified embryos were transferred to 5 recipient gilts, and 17 healthy piglets were produced from 2 recipients (3 recipients aborted) in Group 1. In Group 2, 81 vitrified embryos and 16 fresh embryos in total were transferred to 4 recipient gilts, and 10 healthy piglets from the vitrified embryos were produced from 3 recipients. These results indicated that porcine embryos of compacted morulae to early blastocyst stage can survive cryopreservation using the microdroplet method without any special intracellular manipulation or treatment. PMID- 12749939 TI - Effect on ejaculatory performance and semen parameters of sexually-satiated male goats (Capra hircus) after changing the stimulus female. AB - The association between stimulation of sexual activity and semen characteristics of sexually mature goats was evaluated. Nine 2-year-old criollo bucks were used. Each buck was subjected to 8-weekly trials in which one of the following two treatments was alternately applied. In treatment 1, males were individually exposed to the same estrus-induced female during a 4-h period, and ejaculates were collected and analyzed throughout. Treatment 2 was the same, except that a different doe replaced the stimulus animal for the third and fourth hours. The total number of ejaculations and total sperm number for four sets of data was different (P<0.01) for treatments 1 and 2 (184 versus 252 ejaculations with a total of 354.3 versus 477.1 billion sperm, respectively). In treatment 2, when first and last ejaculation of each male with the (original) stimulus animal were compared, total sperm per ejaculate decreased (P=0.08) from 4.14+/-3.8 x 10(9) to 0.77+/-0.7 x 10(9), while this value increased (P=0.05) to 3.04+/-2.3 x 10(9) after a new female was introduced, which represented a recovery of 67.35%. All males achieved a first ejaculation with the original stimulus animal while on average, only three achieved a seventh service. After changing the stimulus animal, all males ejaculated again. It is concluded that changing the stimulus animal after a 2-h continuous exposure to an estrous female stimulates sexual activity and increases sperm output in male goats. PMID- 12749940 TI - In vitro and in vivo comparison of egg yolk-based and soybean lecithin-based extenders for cryopreservation of bovine semen. AB - Semen diluents containing egg yolk as a cryoprotectant may pose hygienic risks and are difficult to standardize. Although a new generation of semen diluents free of animal ingredients is available, egg yolk-containing extenders are still widely used for cryopreserving semen. We compared the effects of using different extenders on bovine sperm function in vitro and on fertility in vivo. A soy lecithin extender (SL; AndroMed) and an egg yolk-containing (TRIS-EY) extender were tested. No differences (P>0.05) were detected between the two extenders for sperm-zona pellucida binding capacity (HZI=115+/-13). Assessment of the inducibility of the acrosome reaction with progesterone showed no differences (P>0.05) between extenders for live acrosome-reacted sperm (15+/-2.36 and 14.42+/ 2.02%, respectively, for SL and TRIS-EY). However, post-thaw sperm motility was significantly lower (P<0.05) when semen was extended in the TRIS-EY diluent. Field trials revealed that nonreturn rates of SL-extended semen showed significantly higher insemination success (P<0.0001) compared with the nonreturn rates for the TRIS-EY extender (70.45 and 67.85%, respectively). We suggest that consistent with quality standards that should be required for cryoprotectant media and because of the superior quality of the egg yolk-free extender, a defined soybean lecithin-containing diluter might be the better choice as a semen extender in the future. PMID- 12749941 TI - Reproductive season affects inhibitory effects from large follicles on the response to superovulatory FSH treatments in ewes. AB - The main objective of this study was to compare the effect of the presence of large follicles at the start of FSH treatment on the superovulatory response in ewes in the breeding and nonbreeding seasons. A second objective was to verify the effect on the superovulatory response of the presence of a corpus luteum at the start of the FSH treatment during the breeding season. Fifteen ewes in breeding season (October) and 14 in nonbreeding season (May-June) were treated with 40 mg FGA sponges (Chronogest) for 14 days, together with a single dose of 125 microg cloprostenol on Day 12, considering Day 0 as day of progestagen insertion. Superovulatory treatments consisted of eight decreasing doses (1.5 ml x 3, 1.25 ml x 2 and 1 ml x 3) of Ovagen twice daily from 60 h before to 24h after sponge removal. Ovarian structures were assessed by transrectal ultrasonography using a 7.5 MHz linear array probe. Luteal activity at progestagen insertion (Day 0) and presence of corpus luteum and of large follicles at first FSH dose (Day 12) were determined. There were no significant differences between the breeding season and nonbreeding season for ovulation rate (11.6+/-1.4 versus 11.6+/-1.3), number of recovered embryos (8.0+/-1.1 versus 9.6+/-1.3) or number of viable embryos (7.2+/-1.1 versus 5.8+/-1.2). During the breeding season, there were fewer recovered embryos in ewes with a large follicle (> or =6mm) at first FSH dose (6.9+/-1.1 versus 12.3+/-1.8, P<0.05) and fewer viable embryos (5.0+/-1.2 versus 10.5+/-0.5, P<0.05) than in ewes without such a follicle. During the nonbreeding season, however, there were no significant differences between ewes with or without a large follicle for either recovered (9.0+/-2.5 versus 11.3+/-1.2) or viable embryos (6.3+/-2.3 versus 8.1+/-1.2). Analysis of seasonal differences in ewes with a large follicle showed a lower number of recovered embryos in the breeding season (P<0.05) due to a lower recovery rate (65.7% versus 92.3%, P<0.05), since mean number of corpora lutea in response to the FSH treatment was similar (10.9+/-1.3 versus 10.0+/-2.5). These results indicate that, in sheep, the inhibitory effects of large follicles during the nonbreeding season are not as obvious as during the breeding season. PMID- 12749942 TI - Breeding behavior of the American black bear Ursus americanus. AB - By means of videotape, we observed and timed courtship, mating, and refractoriness in the American black bear while confined in a semicaptive environment. Sixty-six courtships resulted in mating. During courtship, the male detected estrus and receptivity in the female. Courtship lasted 22.5+/-2.3s (mean+/-S.E.M.). We videotaped and reviewed 61 matings that included mounts, pelvic thrusts, and flutters. Mating lasted 51.6+/-4.2s. Within these 61 matings, there were 24 pelvic thrusts and 14 flutters observed. Pelvic thrusts averaged 2.3+/-0.3 times, while flutters averaged 11.1+/-2.9 times. Of the 61 courtship mating segments, 28 segments depicted a male refractory period, while 27 segments depicted a female refractory period. The male refractory period lasted 44.9+/ 6.1s during which the male sat and licked his genital region. The female walked, ran away, or turned and attacked the male during her 54.2+/-8.3s refractory period. This is the first time a large number of paired ursid matings has been described and timed. PMID- 12749943 TI - Characterization and localization of alkaline phosphatase in canine seminal plasma and gonadal tissues. AB - Alkaline phosphatase (AP) is a useful indicator of the presence of the sperm-rich (2nd) fraction in the canine ejaculate. Two AP isoenzymes originating from separate genes have been identified in the dog: tissue nonspecific (TNS) and intestinal. Bone, liver, and corticosteroid-induced AP are different isoforms of the TNS and intestinal isoenzymes. Using gel electrophoresis and levamisole inhibition assays, it was determined that seminal plasma AP (SAP) is a unique isoform of canine TNS AP whose glycosylation is distinct from either of the TNS AP isoforms commonly found in canine serum. Using immunocytochemistry, SAP activity was localized to the epididymal and seminiferous tubular epithelium. The ability to distinguish SAP from bone AP, liver AP and corticosteroid-induced AP could be beneficial to the practitioner in determining the quality of a semen sample. PMID- 12749944 TI - Immunization against 5alpha-androstane-3alpha,17beta-diol affects ovarian folliculogenesis in Merino ewes. AB - The 5alpha-reduced androgens have been implicated as antagonists of follicular development. In this experiment, we examined the effect of active immunization against 5alpha-reduced androgen on follicular development in ewes. During the breeding season, cyclic Merino ewes were either actively immunized three times against 5alpha-androstane-3alpha,17beta-diol (3alpha-diol) or served as controls. Six to nine weeks after the last immunization, they were treated with PGF(2alpha) analog (PG, 125mg cloprostenol i.m.) and luteolysis was induced. Fourteen days after the PG treatment, the ewes were either killed (mid-luteal phase) or treated a second time with PG and killed 24h later (early follicular phase). At slaughter, blood samples were collected and ovaries recovered. All CL and follicles larger than 2mm were dissected and their size and appearance were recorded. Follicular fluid was collected and concentrations of estradiol-17beta (E(2)), progesterone (P), androstenedione (A(4)), testosterone (T), dihydrotestosterone (DHT), 5alpha-androstane-3alpha-ol,17beta-one (androsterone: 3alpha-ol) and 3alpha-diol were determined by RIA. Immunization induced antibodies primarily to DHT and its 5alpha-reduced substrates 3alpha-diol and 3alpha-ol but not to E(2), P, A(4) or T. Immunization increased ovulation rate, size of ovulatory follicles and weight of CL. Immunization appeared to increase ovulation rate by decreasing the incidence of atresia in large preovulatory follicles. Regardless of their physiological status follicles contained only low levels of DHT; 3alpha-ol and 3alpha-diol were not detected in most follicles. Immunization did not appear to affect levels of DHT or other steroids in the follicular fluid. In conclusion, the induction of antibodies to 5alpha-reduced androgens increases ovulation rate by enhancing follicular viability during the preovulatory period in ewes. However, this effect is not brought about by the direct immune-neutralization of DHT or its 5alpha-reduced substrates 3alpha-ol and 3alpha-diol at the ovarian level. PMID- 12749946 TI - Effect of heparin on cleavage rates and embryo production with four bovine sperm preparation protocols. AB - Heparin is often added to fertilization media to induce sperm capacitation. However, recent observations from the bovine IVF industry have indicated that heparin may not be necessary to induce sperm capacitation when cryopreserved bovine sperm are separated through Percoll gradients. The objective of these studies was to determine if the addition of heparin to fertilization media was required following separation of frozen-thawed bovine sperm. Experiment 1 was conducted to determine the effect of heparin on cleavage rates and embryo production when using Percoll, BSA or washing sperm separation protocols. Experiment 2 was conducted to determine the effect of heparin on cleavage rates and embryonic development when using a new sperm separation protocol (PureSperm). In Experiment 1, regardless of sperm separation protocol, heparin increased cleavage rates and embryo production (P<0.05). Further, cleavage rates and embryo production were higher for the Percoll procedure compared to either BSA or washing sperm separation protocols (P<0.05). In Experiment 2, there was no difference between Percoll and PureSperm separation protocols on cleavage rates or embryo production (P>0.05). However, heparin increased cleavage rates for both protocols and improved embryo production for the PureSperm protocol (P<0.05). In conclusion, use of heparin in fertilization media improves in vitro embryo production when using cryopreserved bovine sperm. Furthermore, PureSperm can be used as an alternative to Percoll for bovine sperm separation. PMID- 12749945 TI - Estrus synchronization in beef cows: comparison between GnRH+PGF2alpha+GnRH and PRID+PGF2alpha+eCG. AB - The aim of this study was to compare two protocols for estrus synchronization in suckled beef cows over a 2 years period. The population studied consisted of 172 Charolais and 168 Limousin cows from 12 and 14 beef herds, respectively. In each herd, cows were allotted to groups according to parity, body condition score and calving difficulty. Cows in Group 1 (n=174) received PRID on Day-8 with estradiol benzoate (10mg, vaginal capsule), dinoprost on Day-4 (25mg i.m.), eCG on Day 2 (500 IU i.m.). The PRID was removed on Day-2 and cows were inseminated on Day 0, 56 h after PRID was removed. Cows in Group 2 (n=166) received GnRH on Day-10 (100 microg i.m.), dinoprost on Day-3 (25mg i.m.) and GnRH on Day-1 (100 microg i.m.), and were inseminated on Day 0, 16-24h after the last GnRH treatment. Plasma progesterone concentrations were measured to determine cyclicity prior to treatment (Days-20 and -10), to confirm the occurrence of ovulation (Days 0 and 10) and to determine the apparent early pregnancy rate (Days 0, 10 and 24). Pregnancy diagnosis was performed by ultrasonography between Days 35 and 45. The effects of various factors on ovulation, apparent early pregnancy and pregnancy rates were studied using logistic mixed models. There was no significant difference between Groups 1 and 2, respectively, for the cyclicity rate before treatment (80.5% versus 80.1%), for apparent pregnancy rate on Day 24 (62.1% versus 54.8%, P=0.09) and for pregnancy rate on Days 35-45 (53.8% versus 46.3%, P=0.16). Ovulation rate was higher (P<0.01) in Group 1 (90.8%) than in Group 2 (77.1%) and was affected by cyclicity prior to treatment in Group 2 but not in Group 1 (Group 1: 88.2% in anestrous cows versus 91.4% in cyclic cows; Group 2: 45.5% in anestrous cows versus 85.0% in cyclic cows, P interaction=0.05). Apparent pregnancy rates on Day 24 were influenced by the year of study (52.4% versus 68.8%, OR=2.12, P<0.01) and by the cyclicity before treatment (anestrous cows 46.3% versus cyclic cows 61.5%, OR=1.86, P<0.05). Pregnancy rates at 35-45 days were influenced by the year of study (44.2% versus 59.8%, OR=1.92, P<0.01). In conclusion, although pregnancy rates were similar for the two treatments, the combination of GnRH+PGF2alpha+GnRH in suckled beef cows induced a lower rate of ovulation than treatment with PRID+PGF2alpha, particularly in anestrous cows. PMID- 12749947 TI - Ovulation rate after GnRH or PGF2alpha administration in early postpartum dairy cows. AB - The objectives of this study evaluating induction of ovulation in early postpartum dairy cows were to: compare two methods of GnRH (100 mcg) administration (i.m. route and s.c. implant), and determine if prostaglandin F(2alpha) (PGF) causes release of LH or ovulation similar to that reported for GnRH. In trial #1, serum LH peaked at 2h after i.m. administration of GnRH and was declining at 4h. The s.c. GnRH implant also caused an elevation in serum LH at 2 and 4h after treatment, with LH declining at 6h. Serum LH was unchanged in control cows. Experimental treatment caused ovulation in 4 of 14 GnRH i.m. treated cows, 4 of 12 GnRH implanted cows and 0 of 13 control cows. Parity had no effect on LH response but did affect resulting ovulation rate as multiparous cows were more likely to ovulate than were primiparous cows in response to either GnRH treatment. All cows that ovulated had a follicle larger than 12 mm at the time of treatment. In trial #2, serum LH increased as before after i.m. administration of GnRH, however, serum LH was unchanged in cows treated with PGF or saline. Gonadotropin releasing hormone caused more cows to ovulate than did PGF or saline treatments, and GnRH shortened the interval from treatment to the onset of CL function over the PGF treatment; 13.9+/-2.6, 28.2+/-4.1 and 22.3+/-4.1 days for GnRH, PGF and saline, respectively. In summary, there was no difference in the ability of s.c. implantation and i.m. administration of GnRH to cause ovulation. Prostaglandin F(2alpha) did not cause release of LH or ovulation. In 22 early postpartum dairy cows treated with 100 mcg GnRH i.m. in these two trials, nearly all cows (95%) responded with a release of LH but only 45% (10/22) responded with an ovulation and subsequent formation of a CL. PMID- 12749948 TI - Motion characteristics of Murrah buffalo bull spermatozoa in various seasons and its relationship with functional integrity of the plasmallema. AB - Semen was collected from six adult (3.5-7-year-old) Murrah buffalo bulls at weekly intervals for 1 year and evaluated for routine parameters, motion characteristics, reactivity in hypoosmotic solution, and acrosomal and other morphological abnormalities of the spermatozoa. The overall motility (MOT), straight line velocity (VSL), curvilinear velocity (VCL), linearity (LIN), lateral head displacement (ALH) and average path velocity (VAP) were 66.85+/ 2.79%, 26.58+/-0.24 and 107.07+/-1.47 microm/s, 26.91+/-0.01%, 11.19+/-0.09 and 61.78+/-2.79 microm/s, respectively. Significant seasonal variation was observed in sperm kinematics and hypoosmotic swelling (HOS) reactivity. Except for LIN, the mean values of sperm dynamics were higher during summer and rainy season and significantly lower in winter season. Sperm kinematics showed significant (P<0.01) positive correlation (r=0.25-0.60) with plasmallemal integrity. Ejaculates with less than 50% HOS-reactive spermatozoa had significantly lowered MOT, VSL, VCL and VAP as compared to the ejaculates with >50% HOS-positive spermatozoa. No significant difference was observed in sperm kinematics among the ejaculates having 50-70% and >70% HOS-reactive spermatozoa. The trend of motion dynamics of the spermatozoa with respect to HOS reactivity was similar in all the three seasons (summer, rainy and winter). The results indicate that ejaculates having more than 50% of HOS-reactive sperm show a higher magnitude of sperm kinematics compared to ejaculates having less than 50% HOS-positive spermatozoa. PMID- 12749949 TI - Development of rat oocytes following intracytoplasmic injection of sperm heads isolated from testicular and epididymal spermatozoa. AB - The possibility of obtaining normal development of rat oocytes following intracytoplasmic injection of rat sperm heads, obtained by sonicating spermatozoa from testes and epididymides, was evaluated. Irrespective of the source of spermatozoa, sperm heads were successfully injected into approximately 45% of oocytes used; after 9-12h of culture, approximately 55% of injected oocytes still had normal morphology. Of the oocytes injected with testicular sperm heads 45% were activated, with a female pronucleus and a second polar body, but significantly more oocytes (approximately 68%) injected with caput and cauda epididymal sperm heads were activated. Male pronuclear formation was observed in 67-84% of the activated oocytes, with no difference in the proportions among the different sources of sperm heads. When zygotes showing two pronuclei and a second polar body at 10h after injection were cultured in conditions that support development of 1-cell embryos produced in vivo, no embryos derived from testicular sperm heads developed to blastocysts after 120 h of culture. Development of embryos derived from cauda sperm heads was significantly higher at all points of assessment, while embryos from caput sperm showed an intermediate degree of development, compared with embryos from testicular spermatozoa. However, similar proportions (2-4%) of 1-cell embryos derived from all three groups of sperm heads developed into normal offspring after transfer to foster mothers; of the limited number of offspring tested, all were fertile. These results demonstrate that sperm heads from all sources tested are similar in their ability to contribute to full development of normal, fertile offspring. PMID- 12749950 TI - Semen parameters and level of microsatellite heterozygosity in Noriker draught horse stallions. AB - It was the aim of the present study to determine physiological values for different semen parameters in an endangered draught horse breed, the Austrian Noriker. Because small population size is often believed to cause a decrease in fertility and/or semen quality through inbreeding and a reduction in genetic variation, the general genomic heterogeneity of the breed was estimated on the basis of microsatellite variation and correlated to semen parameters. Semen could be collected from 104 of 139 stallions with semen collection being more often successful in younger stallions. Mean volume of ejaculates was 90.8+/-55.1 ml, density 243+/-114 x 10(6)ml(-1), total sperm count 21.0+/-23.7 x 10(9), percentage of morphologically normal spermatozoa 38+/-18% and total motility 50+/ 23%. Total sperm count and semen motility were significantly affected by age. Blood samples of 134 stallions were analysed for 12 microsatellite DNA markers. Genotypes of 110 stallions with at least 11 successfully typed markers were used for calculation of heterozygosity. A total of 82 alleles was identified with a mean of 6.8 alleles per marker. Heterozygosity varied between 35 and 76% for the different markers, mean heterozygosity was calculated to 63%. No correlation between heterozygosity and semen parameters was found. PMID- 12749951 TI - Premature prostaglandin F2alpha secretion causes luteal regression in GnRH induced short estrous cycles in cyclic dairy heifers. AB - This study aimed to confirm that the luteolysis in normal-cycling dairy heifers seen during short estrous cycles induced with cloprostenol (Clp) and GnRH administered 24h apart is caused by a premature release of prostaglandin F(2alpha) (PGF(2alpha)). A further aim was to study the PGF(2alpha) release pattern more closely to determine whether it resembles the spontaneous release occurring during normal regression of the corpus luteum (CL) or whether PGF(2alpha) is continuously secreted after the induced ovulations, leading to short estrous cycles. Twenty-four Ayrshire heifers were allotted to four equally sized groups. After estrus synchronization with 0.5mg of Clp, a new luteolysis was induced with 0.5mg of Clp on Day 6 (groups T-d6 and C-d6) or Day 7 (groups T d7 and C-d7) after ovulation. Gonadorelin (0.1mg i.m.) was given to groups T-d6 and T-d7 to induce premature ovulation 24h later. Groups C-d6 and C-d7 served as controls. Ovaries were examined daily by transrectal ultrasonography, while blood samples (for progesterone and 15-ketodihydro-PGF(2alpha) analyses) were obtained via a jugular catheter every 3h, starting from the second Clp treatment and continuing for 9 days postovulation. Unresponsiveness to Clp or anovulation resulted in 4 C-d6 heifers being excluded. Four heifers in group T-d6 and three in group T-d7 had a short estrous cycle of 8-12 days, while all others had a cycle of normal length. Significant elevations in 15-ketodihydro-PGF(2alpha) concentrations with recurrent high peaks coincided with a decrease in progesterone concentration and were detected in all heifers that showed a short estrous cycle, but not in any heifers with normal estrous cycles in groups T and C. In conclusion, a premature release of PGF(2alpha), which closely resembles its release during spontaneous luteolysis, causes luteal regression in these short cycles. PMID- 12749952 TI - Anger and persistent pain: current status and future directions. PMID- 12749953 TI - Chronic intrathecal cannulas inhibit some and potentiate other behaviors elicited by formalin injection. PMID- 12749954 TI - Age differences in postoperative pain are scale dependent: a comparison of measures of pain intensity and quality in younger and older surgical patients. AB - As the population ages, research into the assessment of postoperative pain in older patients is urgently needed. The reliability and validity of most pain scales for the assessment of acute postoperative pain in the elderly remain to be demonstrated. The present study reports the analysis of age-related patterns on three pain scales (McGill Pain Questionnaire, MPQ; Present Pain Intensity, PPI; and Visual Analog Scale, VAS) completed by younger (n=95, mean age=56.4+/-5.8 years) and older (n=105; mean age=66.8+/-2.7 years) men following radical prostatectomy. All patients received intravenous morphine via patient-controlled analgesia (PCA) throughout the study. On the first 2 postoperative days (POD), patients completed the pain scales and PCA opioid intake was recorded. An interaction was found between amount of opioid self-administered and POD. In both groups, less opioid was administered on POD 2 than POD 1, but the decrease over time was greater in younger than older men. On both PODs, older men self administered less opioid than younger men. Age differences in pain were dependent on the pain scale used. Older men had significantly lower scores than younger men on the MPQ and PPI but there were no differences on the VAS. Several age differences in the psychometric properties of the scales were evident. On both PODs, the correlation between VAS and MPQ scores was significantly lower in the older than younger group. POD effect sizes did not differ between the scales or age groups suggesting that all three scales have comparable sensitivity within an age group. However, the different results between the scales for the effect of age suggests that the VAS is not sufficiently sensitive to detect age differences. Therefore, age differences in postoperative pain are better captured by verbal descriptions of pain qualities than non-verbal measures of intensity. PMID- 12749955 TI - Multiple phases of relief from experimental mechanical allodynia by systemic lidocaine: responses to early and late infusions. AB - Systemic lidocaine can relieve various forms of neuropathic pain that develop after nerve injury. Mechanical allodynia, defined by a significant drop in paw withdrawal threshold force following spinal nerve ligation (L5-L6) in rats, can be reversed by one 30min lidocaine infusion at a constant plasma concentration as low as 1-2 microg/ml, an effect that is still present when the rats are tested days and weeks afterwards. In this study, we resolved the detailed time course of reversal of ipsilateral and contralateral allodynia in rats with spinal nerve ligation by a single systemic infusion of lidocaine, to 4 microg/ml, given either 2 days after ligation (POD2) or 7 days after ligation (POD7). Male Sprague-Dawley rats were examined for 21 days after undergoing sham operation or spinal nerve ligation to produce allodynia, which was quantified by a lower force of von Frey hairs at the plantar hind paw just required to produce paw withdrawal (paw withdrawal threshold, PWT). Six experimental protocols were followed: rats were infused with lidocaine on POD2 (L2) or on POD7 (L7), or with saline on POD2 (S2) or on POD7 (S7), and sham operated rats were infused with lidocaine on POD2 or on POD7. PWTs were measured during the last 5min of a single 30min lidocaine infusion; at 30, 60, 90, 120, 240 and 360min, and 24, 48 and 72h after beginning infusion, and then every 1-3 days up to 21 days. Three distinct sequential phases of ipsilateral relief were apparent in both L2 and L7 groups: (1) an acute elevation of PWT during the infusion, returning to the pre-infusion allodynic level within 30-60min after infusion; (2) a second, transient elevation of PWT within the next 360min; (3) a sustained elevation of PWT developing slowly over 24h after infusion and maintained over the next 21 days. A significant, although weaker contralateral allodynia developed more slowly (>POD8) than the ipsilateral condition, and could be delayed for more than 2 weeks by lidocaine infusion on POD2 but for only 1 week by the same treatment on POD7. None of the sham operated animals had any allodynic signs and no saline infusions elevated PWT in ligated, allodynic rats. These results of separate phases imply that there are mechanistic differences between the acute relief and the sustained relief of allodynia after a single infusion of lidocaine, and may present an experimental paradigm for investigating the advantages of earlier rather than late therapeutic intervention. PMID- 12749956 TI - Assessment of nociceptive trigeminal pathways by laser-evoked potentials and laser silent periods in patients with painful temporomandibular disorders. AB - We assessed the trigeminal nociceptive pathways in patients with painful temporomandibular disorders (TMD) and control subjects using a CO(2)-laser stimulator which provides a predominant activation of the nociceptive system. Fifteen patients with unilateral pain were examined in accordance with the Research Diagnostic Criteria for TMD and 30 gender- and age-matched individuals were included as a control group. Laser-evoked potentials (LEPs) and laser silent periods (LSPs) after stimulation of the perioral region (V2/V3) on the painful and non-painful sides were recorded in all subjects. LEPs were evoked by low intensity pulses (1.5 x perception threshold (PTh)) and recorded from scalp electrodes at the vertex. LSPs were evoked by high-intensity pulses (4 x PTh) and recorded bilaterally from masseter muscles with surface electromyogram (EMG) electrodes. Subjects also assessed the stimulus intensity on a 0-10 rating scale. LEPs had normal latency but smaller amplitude in TMD patients compared to the control group (P<0.001). Side-to-side comparison within patients showed that LEP amplitude was even more reduced after stimulation on the painful than the non painful side (P<0.001). TMD patients showed a significant side-asymmetry of the pre-stimulus EMG activity, with a smaller value in the muscle on the painful side (P<0.001). LSPs were completely absent bilaterally in 12 TMD patients and unilaterally in two patients; only one patient had normal and bilateral LSPs. TMD patients perceived the laser stimulus less intense on the painful than the non painful side (P<0.05). We found suppression of cortical responses and brainstem reflexes elicited by a predominantly nociceptive input in TMD patients. These findings are consistent with recent experimental pain studies and suggest that chronic craniofacial pain in TMD patients may be associated with a dysfunction of the trigeminal nociceptive system. PMID- 12749957 TI - Chronic pain, work performance and litigation. AB - The overall population impact of chronic pain on work performance has been underestimated as it has often been described in terms of work-related absence, excluding more subtle effects that chronic pain may have on the ability to work effectively. Additionally, most studies have focussed on occupational and/or patient cohorts and treatment seeking, rather than sampling from the general population. We undertook a population-based random digit dialling computer assisted telephone survey with participants randomly selected within households in order to measure the impact of chronic pain on work performance. In addition, we measured the association between pain-related disability and litigation. The study took place in Northern Sydney Health Area, a geographically defined urban area of New South Wales, Australia, and included 484 adults aged 18 or over with chronic pain. The response rate was 73.4%. Working with pain was more common (on an average 83.8 days in 6 months) than lost work days due to pain (4.5 days) among chronic pain participants in full-time or part-time employment. When both lost work days and reduced-effectiveness work days were summed, an average of 16.4 lost work day equivalents occurred in a 6-month period, approximately three times the average number of lost work days. In multiple logistic regression modelling with pain-related disability as the dependent variable, past or present pain-related litigation had the strongest association (odds ratio (OR)=3.59, P=0.001). In conclusion, chronic pain had a larger impact on work performance than has previously been recognised, related to reduced performance while working with pain. A significant proportion were able to work effectively with pain, suggesting that complete relief of pain may not be an essential therapeutic target. Litigation (principally work-related) for chronic pain was strongly associated with higher levels of pain-related disability, even after taking into account other factors associated with poor functional outcomes. PMID- 12749959 TI - Aftersensations in experimental and clinical hypersensitivity. AB - This study examined aftersensations after repetitive pinprick stimulation in nerve injury patients with allodynia and in capsaicin sensitized skin in control subjects. Forty-one patients and 12 male control subjects were stimulated repetitively with a computer-driven von Frey hair in sensitized skin and in contralateral normal skin at 0.2 and 2.0Hz. Pain during and after stimulation was scored on an electronic visual analogue scale and sampled on a computer. Maximum evoked pain, time to pain onset, duration of pain and duration of aftersensations were calculated from stored data. Aftersensations were significantly more frequent in sensitized skin than in contralateral normal skin in both control subjects and patients. Pain was evoked more frequently and maximum evoked pain score was significantly higher in sensitized skin compared to contralateral normal skin in both groups. Furthermore, evoked pain score was frequency dependent with a significantly higher pain score at 2.0 than at 0.2Hz. There was a significant positive correlation between maximum evoked pain scores and duration of aftersensations both in patients and control subjects. No difference was observed in maximum pain score, time to pain onset and duration of aftersensations between the groups. Findings suggest that aftersensations and evoked pain are mediated by a common mechanism, which most likely is central. The clinical manifestation of aftersensations can be mimicked in an experimental pain model so aftersensations may be another useful parameter in assessment of central sensitization. PMID- 12749958 TI - Tricyclic antidepressants as long-acting local anesthetics. AB - Amitriptyline, nortriptyline, imipramine, doxepin, desipramine, protriptyline, trimipramine, and maprotiline are tricyclic antidepressants (TCAs) used orally in treating major depressive disorders. Recent studies showed that amitriptyline is more potent in blocking the sciatic nerve functions in vivo by local injection than bupivacaine, a long-acting local anesthetic. We therefore tested whether various TCAs could likewise act as local anesthetics in vivo after single injection via the rat sciatic notch. The duration of complete sciatic nerve blockade by TCAs and the time to reach full recovery were measured with neurobehavioral assays and compared with results from bupivacaine. Amitriptyline, doxepin, and imipramine at 5mM elicited a longer complete sciatic nerve blockade than did bupivacaine at 15.4mM (0.5%), whereas trimipramine and desipramine at 5mM produced a shorter blockade. In contrast, nortriptyline, protriptyline, and maprotiline failed to elicit complete sciatic nerve blockade. Thus, TCAs have very different efficacy as local anesthetics in vivo. The duration of rat sciatic nerve blockade in vivo by TCAs is not well correlated with the 50% inhibitory concentration (IC(50)) of TCAs in blocking human cardiac Nav1.5 Na(+) channels expressed in human embryonic kidney cells. With this in vitro expression system, TCAs appear more potent than bupivacaine as Na(+) channel blockers in Nav1.5 Na(+) channels. We suggest that the ability of TCAs to pass through various membrane barriers within peripheral nerve trunks is crucial to their local anesthetic efficacy in vivo. TCAs with a tertiary amine appear more effective in penetrating these membrane barriers than TCAs with a secondary amine. PMID- 12749960 TI - Development of motor system dysfunction following whiplash injury. AB - Dysfunction in the motor system is a feature of persistent whiplash associated disorders. Little is known about motor dysfunction in the early stages following injury and of its progress in those persons who recover and those who develop persistent symptoms. This study measured prospectively, motor system function (cervical range of movement (ROM), joint position error (JPE) and activity of the superficial neck flexors (EMG) during a test of cranio-cervical flexion) as well as a measure of fear of re-injury (TAMPA) in 66 whiplash subjects within 1 month of injury and then 2 and 3 months post injury. Subjects were classified at 3 months post injury using scores on the neck disability index: recovered (<8), mild pain and disability (10-28) or moderate/severe pain and disability (>30). Motor system function was also measured in 20 control subjects. All whiplash groups demonstrated decreased ROM and increased EMG (compared to controls) at 1 month post injury. This deficit persisted in the group with moderate/severe symptoms but returned to within normal limits in those who had recovered or reported persistent mild pain at 3 months. Increased EMG persisted for 3 months in all whiplash groups. Only the moderate/severe group showed greater JPE, within 1 month of injury, which remained unchanged at 3 months. TAMPA scores of the moderate/severe group were higher than those of the other two groups. The differences in TAMPA did not impact on ROM, EMG or JPE. This study identifies, for the first time, deficits in the motor system, as early as 1 month post whiplash injury, that persisted not only in those reporting moderate/severe symptoms at 3 months but also in subjects who recovered and those with persistent mild symptoms. PMID- 12749961 TI - Differential recruitment of endogenous pain inhibitory systems in neuropathic pain patients. AB - Neuronal hyperexcitability is a key finding in patients with neuropathic pain. Contributing to hyperexcitability may be decreased activity in the endogenous pain inhibitory systems. The present study aimed at recruiting descending inhibition, by the use of painful heterotopic stimulation (HTS), in 16 patients with peripheral chronic neuropathic pain and associated brush-evoked allodynia. Two experiments were performed: one examined the effect of HTS on ongoing pain and intensity of brush-evoked allodynia and the other tested the effect of HTS on ongoing pain and area of brush-evoked allodynia. Both experiments consisted of two sessions, one with painful cold HTS (1 degrees C water bath) another with non painful neutral HTS (32 degrees C water bath). The area of brush-evoked allodynia was significantly reduced (P=0.003) during painful HTS, as compared to non painful HTS. In contrast, neither the intensity of brush-evoked allodynia nor the ongoing pain was significantly changed. The results indicate that endogenous pain modulating systems can alter some aspects of chronic neuropathic brush-evoked allodynia. The differential effect of painful HTS on ongoing pain and area of brush-evoked allodynia suggest that separate mechanisms are involved. PMID- 12749962 TI - Convergence of cutaneous, muscular and visceral noxious inputs onto ventromedial thalamic neurons in the rat. AB - We have recently described a population of neurons in the lateral part of the ventromedial thalamus (VMl), that respond exclusively to noxious cutaneous stimuli, regardless of which part of the body is stimulated. The purpose of the present study was to investigate the convergence of cutaneous, muscular and visceral noxious inputs onto single, VMl neurons in anesthetized rats. VMl neurons were characterized by their responses to Adelta- and C-fiber activation as well as noxious heat applied to the hindpaw. We investigated whether they responded also to colorectal distensions. In an additional series of experiments, we tested the effects of colorectal, intraperitoneal, intramuscular and subcutaneous applications of the chemical irritant mustard oil (MO). The present study shows that a population of neurons located within the thalamic VMl nucleus, carries nociceptive somatosensory signals from the entire body. All these neurons responded to noxious cutaneous and intramuscular stimuli but not to levels of distension that could be considered innocuous or noxious, of the intact and inflammed colon and rectum. Although colorectal distension did not elicit VMl responses, convergence of visceral as well as muscle and cutaneous nociceptors was demonstrated by the increases in ongoing (background) discharges following intracolonic MO. A distinct effect is seen after MO injection into the lumen of the colon: an increase in ongoing activity for 15min but still a lack of effect of colorectal distension. Moreover, following inflammation induced by subcutaneous injections of MO VMl neurons developed responses to both thermal and mechanical innocuous skin stimulation, reminiscent of allodynia phenomena. It is suggested that the VMl contributes to attentional aspects of nociceptive processing and/or to the integration of widespread noxious events in terms of the appropriate potential motor responses. PMID- 12749963 TI - Susceptibility loci for complex regional pain syndrome. AB - An association between HLA-DR13 and patients with complex regional pain syndrome (CRPS) who progressed towards multifocal or generalized tonic dystonia was recently reported. We now report on a new locus, centromeric in HLA-class I, which was significantly associated with a spontaneous development of CRPS, suggesting an interaction between trauma severity and genetic factors conferring CRPS susceptibility. Additionally, an association with the D6S1014 locus was found, supporting the previous finding of an association with HLA-DR13. PMID- 12749964 TI - Central representation of visceral and cutaneous hypersensitivity in the irritable bowel syndrome. AB - We have previously shown that irritable bowel syndrome (IBS) patients have both visceral and cutaneous hyperalgesia. The neural mechanisms of these forms of hyperalgesia were further characterized by comparing cortical processing of both rectal distension (35, 55mmHg) and cutaneous heat nociceptive stimuli (foot immersion in 45 and 47 degrees C water bath) in IBS patients and in a group of healthy age/sex-matched controls. Our approach relied on functional magnetic resonance imaging neuroimaging analyses in which brain activation in age/sex matched control subjects was subtracted from that found in IBS patients. These analyses revealed that both rectal distension and cutaneous heat stimuli evoked greater neural activity in several brain regions of IBS patients in comparison to age/sex-matched control subjects. These include those related to early stages of somatosensory processing (e.g. thalamus, somatosensory cortex) as well as those more related to cognitive and affective processing (insular, anterior cingulate, posterior cingulate, prefrontal cortex). Thus, our results support the hypothesis that hyperalgesia of IBS is manifested by increased somatosensory processing at all cortical levels. This was found to be the case not only for visceral hyperalgesia but also for cutaneous heat hyperalgesia, a likely form of secondary hyperalgesia. Furthermore, visceral and heat hyperalgesia were accompanied by increased neural activity within the same brain structures. These results support the hypothesis that visceral and cutaneous hyperalgesia in IBS patients is related to increased afferent processing in pathways ascending to the brain rather than to selectively increased activity at higher cortical levels (e.g. limbic and frontal cortical areas). PMID- 12749965 TI - Intravenous adenosine alleviates neuropathic pain: a double blind placebo controlled crossover trial using an enriched enrolment design. AB - Adenosine analogs produce analgesic actions in nociceptive paradigms and alleviate manifestations of neuropathic pain in nerve injury models in rodents. In humans, previous work indicates an analgesic effect for adenosine administered intravenously in postoperative and neuropathic pain. In this double blind placebo controlled crossover trial, we used an enriched enrolment design to determine the effects of intravenous adenosine (50 microg/kg/min over 60min) on neuropathic pain. In Phase 1 of the trial, adenosine was administered in an open label manner, while in Phase 2 adenosine was administered in a double blind placebo controlled manner to 23 adenosine responders who had experienced a 30% or greater response in the open trial. Outcome measures included the McGill pain questionnaire (MPQ), which generates a pain rating index (PRI), and contains a visual analog scale (VAS) of pain intensity, the neuropathy pain scale (NPS), and a VAS for pain relief. Subjects also graded the degree of allodynia and hyperalgesia using a VAS. Adenosine led to a significant reduction in spontaneous pain according to the MPQ-PRI, the MPQ-VAS and the VAS for pain relief. The NPS showed a pattern similar to the MPQ-PRI, with statistically significant reductions in scales 1 (intensity), 3 (hot), 6 (sensitive), 7 (itchy) and 9 (unpleasant). Adenosine also led to a significant reduction in pinprick hyperalgesia, but not in allodynia. Three patients from Phase 1 of the trial experienced long term resolution of their pain following intravenous adenosine (5,16,25 months). The results of this study support previous reports that indicate intravenous adenosine alleviates neuropathic pain and hyperalgesia. PMID- 12749967 TI - Changes in sensory processing in the spinal dorsal horn accompany vincristine induced hyperalgesia and allodynia. AB - Abnormal sensation and pain are major dose-limiting factors in cancer chemotherapy with vincristine. In this study, we have adapted a model of this condition by using repeated daily intraperitoneal injections of vincristine in rats. Mechanical allodynia and hyperalgesia without change in responses to thermal stimuli were first observed following 5-8 days of vincristine treatment (0.1mg/kg/day) and then persisted throughout the remainder of the treatment interval (2-3 weeks). Electrophysiological recording from wide dynamic range (WDR) neurons in the lumbar (L4-L5) spinal dorsal horn in hyperalgesic rats demonstrated significantly increased spontaneous activity and after-discharges to noxious mechanical stimuli (von Frey filaments with a bending force greater than 58.02mN, skin compression 1.3 and 3N, 1mm(2)), increased acute A- and C-fiber responses, after-discharges and abnormal 'wind-up' to electrical stimuli (5mA, 2ms) at 0.1Hz applied across the receptive field. These results suggest a state of central sensitization develops in spinal WDR neurons with repeated vincristine treatment that contributes to the spontaneous pain and hyperalgesia seen in patients and the hyperresponsiveness to sensory stimuli seen in animals treated with vincristine. PMID- 12749966 TI - Analgesia by electrostimulation of the trigeminal ganglion in patients with trigeminopathic pain: a PET activation study. AB - Electrostimulation of the trigeminal ganglion (TGES) has shown good results in treatment of trigeminopathic pain in selected patients. To map the mechanisms of TGES analgesia, we determined changes in relative regional cerebral blood flow (rCBF) in ten patients with trigeminopathic pain using positron emission tomography. The patients were scanned before stimulation (habitual pain), after short-term stimulation (1 min, stTGES) and after long-term stimulation (ltTGES). Highly significant pain alleviation was reported after ltTGES. Relative rCBF changes after stTGES, which was without significant pain relief, were attributed mainly to intrinsic TGES effects. A statistical comparison of the subtraction images of ltTGES and stTGES disclosed significant rCBF increases after ltTGES in rostral parts of anterior cingulate cortex (ACC) and neighboring orbitofrontal and medial frontal cortices. Regression analysis of rCBF changes and subjective ratings of pain revealed an inverse relationship in the ipsilateral rostral ACC, and only rCBF changes in the caudal part of the contralateral ACC were consistent with the encoding of pain. The present study provides evidence for a pain modulating role of the rostral ACC, critically important in electrostimulation induced analgesia, and identifies the caudal ACC as a region encoding pain sensation. PMID- 12749968 TI - The effects of high- and low-intensity percutaneous stimulation on nitric oxide levels and spike activity in the superficial laminae of the spinal cord. AB - Nitric oxide (NO) was measured using a new electrochemical method with a carbon fibre microelectrode at depths of up to 400 microm in the lumbar dorsal horn of the anaesthetised rat. The method allowed extracellular spike recording from single units together with the electrochemical recording at the same electrode. Thirty-six cells with low threshold cutaneous (brush/touch) or wide dynamic range receptive fields (brush/touch plus pinch) were studied. Adequate stimulation of the receptive fields did not alter the extracellular NO level for any cells. Percutaneous needle electrodes inserted into the receptive fields were used to stimulate the cells electrically. Twenty-one cells were stimulated using 10 mA current with 0.05 ms duration (low intensity) pulses to stimulate predominantly A fibre afferents. Single shock stimuli gave short latency spike responses but no change in nitric oxide level. Tetanic bursts of stimuli (400 stimuli at 50 Hz) generated a burst of spikes (spike count 548+/-42) and a transient increase in NO (2.61+/-0.11 microM NO). Nitric oxide synthesis inhibition with N(G)-nitro-L arginine methyl ester (L-NAME) nearly abolished the stimulus-evoked increase in nitric oxide and increased the response of the cells (spike count 694+/-34). However, the inhibition of nitric oxide synthesis had no effect on the receptive fields. Fifteen cells were stimulated with shocks using 5 ms pulses (high intensity), to recruit C-fibre afferents into the input volley. This more intense stimulation increased the evoked NO release to 3.63+/-0.15 microM and the spike response to 647+/-54 in control conditions. Following L-NAME, the evoked NO release was reduced and the evoked spike response was significantly decreased. These results show that tetanic activity in afferent fibres increases NO synthesis in the dorsal horn and that inhibition of nitric oxide synthesis may be associated with a selective attenuation of the spike responses to C-fibre inputs. NO may be necessary to maintain proper function of C-fibre afferent synapses when they are subjected to sustained or tetanic inputs. PMID- 12749969 TI - The social context of gastrointestinal cancer pain: a preliminary study examining the relation of patient pain catastrophizing to patient perceptions of social support and caregiver stress and negative responses. AB - A number of studies have shown that catastrophizing is an important predictor of pain and disability in persons having persistent pain conditions. The newly developed communal model of catastrophizing maintains that catastrophizing is a part of broader, interpersonal style of coping in which coping efforts are directed at interpersonal goals, rather than solely at pain reduction. This study examined the potential interpersonal correlates of pain catastrophizing in a sample of 70 patients having gastrointestinal cancers and their caregivers. Measures of pain catastrophizing, perceptions of social support, pain level, and pain behavior were obtained from patients. Caregivers completed measures that included their judgments about the patient's pain level, caregiver stress, and their tendency to engage in negative responses (critical or avoidant behaviors). Overall, patients who engaged in catastrophizing reported receiving higher levels of instrumental support. Caregivers of patients who catastrophized, rated the patient as having more pain and engaging in more pain behavior. Caregivers of patients who catastrophized, also reported higher levels of caregiver stress and critical behaviors. Taken together, these preliminary findings suggest that pain catastrophizing has interpersonal correlates and support the need for additional research examining the social context of pain catastrophizing. PMID- 12749970 TI - The self-efficacy of family caregivers for helping cancer patients manage pain at end-of-life. AB - This preliminary study examined the self-efficacy of family caregivers with regard to helping cancer patients manage pain at end of life. A sample of 63 family caregivers of hospice-eligible cancer patients with pain provided ratings of their self-efficacy in assisting the patient in pain management and rated their own mood and level of caregiver strain. Patients completed measures of pain and quality of life. Data analyses revealed that caregivers who rated their self efficacy as high reported much lower levels of caregiver strain as well as decreased negative mood and increased positive mood. Caregiver self-efficacy in managing the patient's pain was related to the patient's physical well-being. In dyads where the caregiver reported high self-efficacy, the patient reported having more energy, feeling less ill, and spending less time in bed. Considered overall, the results of this study suggest that caregiver self-efficacy in pain management is important in understanding how caregivers adjust to the demands of caring for cancer patients who have pain at the end of life. PMID- 12749971 TI - Involvement of the anterior pretectal nucleus in the control of persistent pain: a behavioral and c-Fos expression study in the rat. AB - The anterior pretectal nucleus (APtN) participates in nociceptive processing and in the activation of central descending mechanisms of pain control. In this study we used behavioral tests (incisional pain and carrageenan-induced inflammatory pain) and c-Fos expression changes to examine the involvement of the APtN in the control of persistent pain in rats. A 1cm longitudinal incision through the skin and fascia of the plantar region (large incision), or a 0.5cm longitudinal incision through the skin only (small incision) was used, and the postoperative incisional allodynia was evaluated with von Frey filaments. The hyperalgesia produced by the intraplantar administration of carrageenan (25 or 50 microg/100 microl) into a hind paw was evaluated by a modified paw pressure test. The electrolytic lesion of the contralateral, but not ipsilateral, APtN significantly intensified the allodynia produced by a large incision of the hind paw. The incisional allodynia and the carrageenan-induced hyperalgesia were intensified by the microinjection of 2% lidocaine into the contralateral, but not ipsilateral APtN, the effect being significantly stronger when a large incision or a higher carrageenan concentration was utilized. A significant increase in the number of c Fos positive cells was found in the ipsilateral, and mainly in the contralateral APtN of rats submitted to a large incision. The number of positive cells in the superficial or deep laminae of the contralateral spinal cord of control and incised rats was not significantly different. Positive cells in the superficial or deep laminae of the ipsilateral spinal cord were significantly more numerous than in control, the effect being significantly more intense in rats with large incision. The microinjection of 0.5% bupivacaine into the APtN contralateral to the incised hind paw reduced the number of positive cells bilaterally in the APtN, but the effect was significant in the contralateral nucleus only. The number of positive cells in the superficial and deep laminae of the contralateral spinal cord of incised and non-incised animals was not significantly changed by the neural block of the contralateral APtN. In the ipsilateral spinal cord, the incision-induced increase in the number of positive cells was significantly reduced in the superficial lamina and significantly increased in the deep lamina of animals previously treated with bupivacaine in the contralateral APtN. In conclusion, the integrity of the APtN is necessary to reduce the severity of the responses to persistent injury. The results also are in agreement with the current notion that persistent noxious inputs to the APtN tonically activate a descending mechanism that excites superficial cells and inhibits deep cells in the spinal dorsal horn. PMID- 12749973 TI - Measuring pain in children with cognitive impairment: pain response to surgical procedures. AB - This study investigated post-surgical pain in children with profound cognitive impairment (PCI), searching for a core set of cues these children use to express their pain. Fifty-two children were observed while they were admitted to the Sophia Children's Hospital for surgery, twice before and five times after surgery. All observations were scored with the item pool consisting of 134 possible pain indicators, using a five-point scale ranging from 0 (never shown) to 4 (always shown). Second, we used the visual analogue scale (VAS) to give a general impression of the severity of the children's pain during the episodes they were observed. Several analyses provided evidence that 23 observable behaviors are sensitive to post-surgical pain in children with PCI, regardless of the pain intensity of the surgical procedures they underwent. The finding that all indicators, except for one, were scored significantly higher on episodes with VAS ratings >or=4, indicates the sensitivity of these indicators concerning absence versus presence of clinically meaningful levels of pain. This study reveals the potential clinical utility of a core set of indicators which can be used to assess post-surgical pain in children with PCI. PMID- 12749972 TI - A cannabinoid agonist differentially attenuates deep tissue hyperalgesia in animal models of cancer and inflammatory muscle pain. AB - Pain associated with cancer and chronic musculoskeletal disorders can be difficult to control. We used murine models of cancer and inflammatory muscle pain to examine whether the cannabinoid receptor agonist WIN55,212-2 reduces hyperalgesia originating in deep tissues. C3H/He mice were anesthetized and implanted with osteolytic NCTC clone 2472 cells into the humeri or injected with 4% carrageenan into the triceps muscles of both forelimbs. At the time of peak hyperalgesia, WIN55,212-2 (1-30mg/kg) or vehicle was administered intraperitoneally and forelimb grip force was measured 0.5-24h later. WIN55,212-2 produced time- and dose-related antihyperalgesia in both models. A 10mg/kg dose of WIN55,212-2 fully reversed carrageenan-evoked muscle hyperalgesia. However, 30mg/kg of WIN55,212-2 attenuated tumor-evoked hyperalgesia only approximately 50%. After controlling for the difference in magnitude of hyperalgesia between the two models, WIN55,212-2 was still more potent at reducing hyperalgesia in the inflammatory model. In the cancer pain model, the antihyperalgesic effect of WIN55,212-2 was partially blocked by pretreatment with the selective CB1 (SR141716A) but not the CB2 (SR144528) receptor antagonist. In contrast, both antagonists blocked antihyperalgesic effects of WIN55,212-2 on carrageenan-evoked muscle hyperalgesia. Catalepsy and loss of motor coordination, known side effects of cannabinoids, did not account for the antihyperalgesia produced by WIN55,212 2. These data show that cannabinoids attenuate deep tissue hyperalgesia produced by both cancer and inflammatory conditions. Interestingly, cannabinoids differentially modulated carrageenan- and tumor-evoked hyperalgesia in terms of potency and receptor subtypes involved suggesting that differences in underlying mechanisms may exist between these two models of deep tissue pain. PMID- 12749974 TI - Complex regional pain syndrome type I: incidence and prevalence in Olmsted county, a population-based study. AB - The objective of this study is to undertake a population based study on the incidence, prevalence, natural history, and response to treatment of complex regional pain syndrome (CRPS). All Mayo Clinic and Olmsted Medical Group medical records with codes for reflex sympathetic dystrophy (RSD), CRPS, and compatible diagnoses in the period 1989-1999 were reviewed as part of the Rochester Epidemiology Project. We used IASP criteria for CRPS. The study population was in the Olmsted County, Minnesota (1990 population, 106,470). The main outcome measures were CRPS I incidence, prevalence, and outcome. Seventy-four cases of CRPS I were identified, resulting in an incidence rate of 5.46 per 100,000 person years at risk, and a period prevalence of 20.57 per 100,000. Female:male ratio was 4:1, with a median age of 46 years at onset. Upper limb was affected twice as commonly as lower limb. All cases reported an antecedent event and fracture was the most common trigger (46%). Excellent concordance was found between symptoms and signs and vasomotor symptoms were the most commonly present. Three phase bone scan and autonomic testing diagnosed the condition in >80% of cases. Seventy-four percent of patients underwent resolution, often spontaneously. CRPS I is of low prevalence, more commonly affects women than men, the upper more than the lower extremity, and three out of four cases undergo resolution. These results suggest that invasive treatment of CRPS may not be warranted in the majority of cases. PMID- 12749975 TI - Facet joint injection: a rare form cause of epidural abscess formation. AB - The posterior zygo-apophyseal joints (facet joints) may be a significant source of back pain. Invasive treatment typically consists of injecting the joints with local anaesthetic and steroid or by radiofrequency ablation of the nerve supply to the joint. Facet joint injection is generally considered to be a very safe procedure with few significant side effects reported. Epidural abscess is a rare but potentially very serious occurrence. Most cases occur spontaneously but the condition may complicate epidural anaesthesia, spinal anaesthesia or epidural steroid injection. We report a case in which facet joint injections resulted in epidural abscess formation. To our knowledge this has not previously been reported. PMID- 12749976 TI - Cervical transforaminal injection of corticosteroids into a radicular artery: a possible mechanism for spinal cord injury. AB - Spinal cord injury has been recognized as a complication of cervical transforaminal injections, but the mechanism of injury is uncertain. In the course of a transforaminal injection, an observation was made after the initial injection of contrast medium. The contrast medium filled a radicular artery that passed to the spinal cord. The procedure was summarily abandoned, and the patient suffered no ill effects. This case demonstrates that despite using careful and accurate technique, it is possible for material to be injected into a radicular artery. Consequently, inadvertent injection of corticosteroids into a radicular artery may be the mechanism for spinal cord injury following transforaminal injections. This observation warns operators to always perform a test injection of contrast medium, and carefully check for arterial filling using real-time fluoroscopy with digital subtraction. PMID- 12749977 TI - Encapsulation of an intrathecal catheter. AB - A 47-year-old patient with cancer pain underwent implantation of an intrathecal drug delivery device. When the patient suffered from an infection with fever, pain on injection into the catheter and an elevated number of granulocytes in the cerebrospinal fluid 7 weeks later, radiologic examination showed an encapsulation of the catheter tip. Concentrations of morphine and morphine-6-glucuronide in the cerebrospinal fluid suggested transport of morphine into the systemic circulation via the vascularisation of the encapsulating membrane. After antibiotic therapy and removal of the catheter, morphine was administered intravenously with a one to one conversion ratio. PMID- 12749979 TI - Spinal cord stimulation for central pain. PMID- 12749978 TI - Diabetic thoracic radiculopathy: an unusual cause of post-thoracotomy pain. AB - Persistent pain is common following thoracotomy. A 64-year-old retired electrician with Type 2 diabetes presented with chest wall and abdominal pain 3 months following video-assisted thoracoscopic surgery (VATS). Postoperatively the patient had suffered pain despite a functioning thoracic epidural catheter. Following investigation, his persistent pain was due to diabetic thoracic radiculopathy (DTR). The disorder is characterized by pain, sensory loss, abdominal and thoracic muscle weakness in patients with diabetes. As in this patient, the pain and sensory loss usually resolve within one year after onset. The disorder may be distinguished from intercostal neuralgia based upon clinical and electromyographic features. PMID- 12749981 TI - Comment on: Serpell et al., gabapentin in neuropathic pain syndromes: a randomised double-blind, placebo controlled trial (Pain 2002; 99: 557-66). PMID- 12749983 TI - Melatonin exerts an antidepressant-like effect in the tail suspension test in mice: evidence for involvement of N-methyl-D-aspartate receptors and the L arginine-nitric oxide pathway. AB - This study investigated the effect of melatonin in the mouse tail suspension test (TST), and the contribution of N-methyl-D-aspartate (NMDA) receptors and the L arginine-nitric oxide (NO) pathway to its antidepressant-like effect. The immobility time in the TST was reduced by melatonin given either by intraperitoneal (0.1-30 mg/kg) or intracerebroventricular (0.001-0.1 nmol/site) route. The anti-immobility effect of melatonin (1 mg/kg, intraperitoneal, i.p.) was prevented by pre-treatment with guanosine 5'-monophosphate (GMP), ascorbic acid, L-arginine or S-nitroso-N-acetyl-penicillamine, but not with D-arginine. Pre-treatment with melatonin (100 mg/kg, i.p.) prevented the anti-immobility effect of MK-801, ketamine or zinc chloride, but did not alter the effect of imipramine. Furthermore, a sub-effective dose of melatonin (0.001 mg/kg, i.p.) produced a synergistic antidepressant-like effect with MK-801, ketamine, zinc chloride and imipramine in the TST. Taken together these data indicate that the effect of melatonin in the TST seems to be mediated through an interaction with NMDA receptors and the L-arginine-NO pathway. PMID- 12749984 TI - Microarray analysis reveals induction of heat shock proteins mRNAs by the torsion dystonia protein, TorsinA. AB - An in-frame deletion (Delta E302/303) in the TorsinA gene has been demonstrated to be responsible for primary torsion dystonia, showing dominant inheritance with reduced penetrance. The Delta E302/303 torsinA mutation forms intracellular ER derived inclusions in a variety of cultured cells, which may suggest that the mutations might evoke ER stress. We used microarray analysis of human derived cell lines expressing the Delta E302/303 torsinA mutation in order to reveal alterations in gene expression in the hope of identifying genetic modifying loci or novel markers for disease pathogenesis. We identified transcriptional changes in multiple members of the heat shock protein family of genes, confirmed by reverse transcription-polymerase chain reaction, which could be indicative of ER stress. However, both wild type and mutant torsinA were affected to a similar extent, suggesting that this is not related to either disease state or the formation of ER-derived inclusions. PMID- 12749985 TI - Chloral hydrate and ethanol, but not urethane, alter the clearance of exogenous dopamine recorded by chronoamperometry in striatum of unrestrained rats. AB - Several general anesthetics reduce dopamine (DA) neuronal activity and release. However, anesthetic-induced alterations in DA transporter (DAT) function are unclear. We used high-speed chronoamperometry to examine how urethane, chloral hydrate and ethanol affected clearance of locally-applied DA in the dorsal striatum of unrestrained rats. Maximal DA signal amplitudes (A(max)) and signal decay rate constants (k) were monitored continuously from the awake to anesthetized state. Urethane did not significantly change A(max) or k. Chloral hydrate and ethanol potentiated A(max) by 32% and 37%, respectively, over baseline, indicative of DAT inhibition. These effects were not additive. Only ethanol increased k, an effect blocked by chloral hydrate. These results suggest differential effects of anesthetics on striatal DAT function, with chloral hydrate and ethanol significantly altering DAT activity but urethane producing only minimal effects. PMID- 12749986 TI - Effect of peripheral cholecystokinin receptor agonists on c-Fos expression in brain sites mediating food consumption in rats. AB - Peripheral cholecystokinin (CCK) elicits satiety by acting on hypothalamic nuclei. The anoretic effect of CCK is mediated by the vagus nerve and involves brainstem areas receiving vagal inputs, such as the nucleus tractus solitarius (NTS) and the area postrema (AP). This work aims to analyze, by measuring c-Fos expression, the effect of selective CCK receptor agonists on brain areas involved in food-intake/satiety process. We observed that SR-146,131, a CCK(1)R agonist, increased c-Fos expression in NTS and AP as well as in some hypothalamic nuclei. CCK-4, a CCK(2)R agonist which does not cross the blood-brain barrier (BBB), only was effective in the hypothalamus. Our data show that the activation of the brainstem is not a requisite to obtain a hypothalamic effect of peripheral CCK and suggest that CCK-4 may indirectly stimulate hypothalamic areas endowed with BBB, without previous activation of neither NTS nor AP. PMID- 12749987 TI - Rapid induction of serotonergic hyperinnervation in the adult rat striatum with extensive dopaminergic denervation. AB - The aim of our study was to determine whether serotonergic hyperinnervation is rapidly induced in the striatum of adult rats with extensive dopaminergic denervation. Immunohistochemical study was performed on the brain sections obtained at 2 and 8 weeks after injection of 6-hydroxydopamine (6-OHDA) into the medial forebrain bundle. The extent of dopaminergic denervation was evaluated as a percentage loss of tyrosine hydroxylase immunopositive neurons in the substantia nigra pars compacta. The immunopositive areas for serotonin (5-HT) in the striatum were measured. In the lesioned rats 97.5+/-0.6% of dopamine neurons were lost. 5-HT immunopositive areas in the striatum were significantly increased both at 2 and 8 weeks after 6-OHDA injection (P<0.01). These results suggest that extensive dopaminergic denervation in adult rats induces rapid serotonergic hyperinnervation in the striatum as early as 2 weeks after lesioning. PMID- 12749988 TI - Aspirin selectively augmented N-methyl-D-aspartate types of glutamate responses in cultured spiral ganglion neurons of mice. AB - Aspirin is commonly used to study tinnitus in animal models because of its ability to induce tinnitus in human subjects. However, the mechanism by which aspirin affects auditory function remains unclear. To investigate the effect of aspirin on the cochlear neurotransmission, we studied its interactions with major types of membrane channels and receptors regulating the excitability of cultured type I spiral ganglion (SG) neurons. Results showed that aspirin had little effect on voltage-gated sodium and potassium currents of SG neurons. In contrast, it selectively potentiated the N-methyl-D-aspartate (NMDA) subtype of the glutamate responses in SG neurons while showing little effect on the alpha-amino 3-hydroxy-5-methylisozazole-4-propionic acid and kainate types of glutamate responses. The aspirin-induced current in the presence of NMDA increased in a dose-dependent manner with a half maximal concentration of 2.2 mM, and it was blocked by NMDA receptor antagonist 2-amino-5-phosphonopentanoic acid or Mg(2+). These in vitro results suggested that aspirin could interfere with the glutamatergic neurotransmission in the cochlea by selectively amplifying NMDA mediated responses. PMID- 12749989 TI - Bacterial lipopolysaccharide shifts fasted plasma ghrelin to postprandial levels in rats. AB - Lipopolysaccharide (LPS) injected intraperitoneally (i.p.) is known to decrease food intake. Ghrelin is a peptide hormone produced by the stomach with a potent orexigenic effect and plasma levels that are inversely correlated with the fed state. We examined changes in plasma ghrelin levels 3 h after LPS (100 microg/kg, i.p.) in fasted rats with or without a 1 h re-feeding period. LPS injection decreased the fasting levels of ghrelin by 51+/-5% compared with preinjection values while i.p. vehicle did not modify ghrelin levels in fasted rats. LPS at this dose reduced fasting-induced food intake by 60% compared with the i.p. vehicle group. Re-feeding decreased plasma ghrelin levels by 58+/-3% compared with pre-feeding fasting values in i.p. vehicle group. These data provide the first evidence that LPS shifts fasting ghrelin levels to those observed postprandially. PMID- 12749990 TI - Different binding pattern of antibodies to prion protein on lymphocytes from patients with sporadic Creutzfeldt-Jakob disease. AB - In Creutzfeldt-Jakob disease (CJD), progressive neuronal cell death probably occurs as a result of a change in conformation of the physiological prion protein (PrP(C)). There is evidence of participation of the lymphatic system and in particular of lymphocytes in the intracorporeal transportation of the pathological prion protein (PrP(Sc)) in new variant CJD and scrapie. Using fluorescence cytometry, we investigated a possible alteration of PrP(C) on lymphocytes of patients with sporadic CJD. We demonstrated a significantly lower binding pattern of antibodies (3F4) against physiological prion protein to lymphocytes of patients with sporadic CJD (n=16) compared with control patients. In contrast this difference was not found on platelets (n=23). For the first time we were able to present a measurable difference of antibody binding on lymphocytes of patients with CJD. One interpretation of this finding is that lymphocytes patrolling the brain bind and transport PrP(Sc) which has a lower binding affinity for the antibodies directed against physiological PrP. PMID- 12749991 TI - Suppression of lipopolysaccharide-dependent tumor necrosis factor alpha induction in rat microglia, in which protein kinase C alpha is down-regulated. AB - Microglia are thought to produce a cytotoxic cytokine, tumor necrosis factor alpha (TNF alpha), in the pathological brain. Thus, the mechanism that suppresses the induction of TNF alpha in microglia is of interest. We found that lipopolysaccharide (LPS)-induced TNF alpha is strongly suppressed by pretreatment with specific protein kinase C (PKC) inhibitors, Go6976 and bisindolylmaleimide, suggesting that PKC alpha plays a role in the signaling cascade of TNF alpha induction. Therefore, the association of PKC alpha with TNF alpha induction was investigated in PKC alpha-deficient microglia prepared by pretreatment with phorbol-12-myristate-13-acetate (PMA). PMA-pretreated microglia strongly decreased the TNF alpha induction in the absence of PKC alpha when activated with LPS. The lack of PKC alpha in the PMA-treated microglia was confirmed by determining conventional PKC activity. Taken together, the present study demonstrated that PKC alpha is a requisite PKC isoform in the signaling cascade of TNF alpha induction in LPS-stimulated microglia. PMID- 12749992 TI - Reverse microdialysis of ionotropic glutamate receptor agonists in the rat globus pallidus increased extracellular dopamine. AB - The globus pallidus (GP) receives a prominent dopamine (DA) input by collaterals from the nigrostriatal pathway. However, the neurochemical regulation of extracellular DA in the GP is unknown at present. In the present study we investigated whether activation of ionotropic glutamate receptors by reverse microdialysis of NMDA and AMPA elicits changes in extracellular DA in the GP. Results show that reverse microdialysis of 70 microM AMPA and of 1 mM NMDA produced a significant and reversible increase of dialysate DA levels to about 130% and 160%, respectively. These data provide evidence that ionotropic glutamate receptors in the GP exert excitatory actions on DA release which might be brought about by actions on presynaptic ionotropic glutamate receptors in the GP or involve stimulation of midbrain DA neurons through trans-synaptic pathways. PMID- 12749993 TI - Evidence that nitric oxide regulates the acute effects of ethanol on rat N-methyl D-aspartate receptors in vitro. AB - Acute exposure to ethanol has been shown to inhibit the function of N-methyl-D aspartate (NMDA) receptors (NMDAR). However, the mechanism by which ethanol produces inhibition of NMDAR and the factors that modulate this effect are not completely understood. Nitric oxide (NO) is an important modulator of NMDAR function in the hippocampus. Therefore, we examined the effects of NO donors on the ethanol-induced inhibition of NMDAR. Primary cultures of hippocampal neurons were prepared from postnatal day 3 rats. After 7 days in culture, NMDAR currents were recorded by using whole-cell patch-clamp electrophysiological techniques. Effects of acute exposure to ethanol on these currents were assessed in the absence and presence of NO donors. We found that the NO donors 1-hydroxy-2-oxo-3 (N-ethyl-2-aminoethyl)-e-ethyl-1-triazene (NOC-12, 100 microM) and S-nitroso-N acetyl-D,L-penicillamine (SNAP, 500 microM) inhibit currents gated by 100 microM NMDA plus 0.5 microM glycine. The inhibitory effect of NOC-12 on NMDAR currents could not be observed when 100 microM of the NO scavenger 2-phenyl-4,4,5,5 tetramethylimidazoline-1-oxyl-3-oxide (PTIO) was present. Importantly, it was found that ethanol inhibits NMDAR responses to a significantly lesser extent in the presence of these donors. Ethanol (65 mM) inhibited NMDAR responses by 42+/ 2%. In the presence of NOC-12 or SNAP, ethanol inhibited NMDAR responses by 21+/ 4% and 11+/-7%, respectively. The effect of NOC-12 on ethanol's actions on NMDAR currents was blocked by PTIO. Our results suggest that NO is a novel modulator of the acute effects of ethanol on NMDAR function. PMID- 12749994 TI - The effects of dexmedetomidine and halothane on Fos expression in the spinal dorsal horn using a rat postoperative pain model. AB - We investigated the effect of an intrathecal injection of a selective alpha2 adrenergic receptor agonist, dexmedetomidine (Dex), and halothane anesthesia on Fos expression in the lumbar spinal dorsal horn after skin incision of the plantar surface of the hind paw, a postoperative pain model using rats. Fos immunoreactivity was induced particularly in the superficial layers of the spinal cord 2 h after surgery. Halothane anesthesia (0.5-1.5%) partially reversed Fos induction, but not in a dose-dependent manner. Preoperative spinal Dex (0.1-10 microg) dose-dependently reduced Fos immunoreactivity, while a relatively high dose of Dex (10 microg) was necessary to produce a profound effect. When used with halothane anesthesia, relatively low doses of Dex (1-3 microg) completely suppressed Fos induction in the superficial spinal layers. These findings indicate that preoperative Dex treatment may provide anesthesia that does not induce stress on spinal neurons. PMID- 12749995 TI - Acupuncture increases neuropeptide Y expression in hippocampus of maternally separated rats. AB - Maternal separation (MS) is a risk factor in the development of mood-related disorders such as depression. Human and animal studies support the involvement of neuropeptide Y (NPY) in the pathology of depression. To investigate the effect of acupuncture on depression-like behavior and examine changes in NPY expression associated with MS, we observed body weight and locomotor activity, and performed NPY immunohistochemistry in the hippocampus. MS for 7 days beginning on postnatal day 14 induced a significant decrease in body weight and locomotion, while acupuncture treatment at acupoint Shenmen (HT7) resulted in a significant increase in both. NPY-immunoreactive cells in area CA1 and the dentate gyrus were decreased in the MS group, but significantly increased in the acupuncture group. These findings suggest that acupuncture has an effect on the depression-like disorder caused by MS, possibly by modulating NPY expression in the hippocampus. PMID- 12749996 TI - Evoked prefrontal gamma oscillation by hippocampal train stimulation in anesthetized rats. AB - We previously reported a difference in short-term synaptic plasticity between the rat posterior dorsal CA1 (pdCA1)-prefrontal cortex (PFC) and ventral CA1 (vCA1) PFC pathways. Here, to determine the effects of hippocampal train stimulation on the local field potential in the medial PFC, we recorded the PFC field potential with brief 250-Hz train stimulation (1, 3, 5, 7, and 9 pulses) of pdCA1 or vCA1 in anesthetized rats. Analysis of the gamma-band (40-100 Hz) power revealed stimulation-evoked gamma oscillation in the pdCA1-PFC, but not in the vCA1-PFC. These results indicate that these pathways have different responses to train stimulation. The function of the pdCA1-PFC may differ from that of the vCA1-PFC, a well-known hippocampus-PFC pathway. PMID- 12749997 TI - Low efficacy adenosine A1 agonists inhibit striatal acetylcholine release in rats improving central selectivity of action. AB - The objective of this study was to characterize the effects of the adenosine A(1) receptor agonist N(6)-cyclopentyladenosine (CPA) and its low efficacy derivatives 2'-deoxy-CPA (2DCPA), 3'-deoxy-CPA (3DCPA), 8-ethylamino-CPA (8ECPA) and 8 butylamino-CPA (8BCPA) on the release of acetylcholine (ACh) using intrastriatal microdialysis. These low efficacy agonists exhibited lower effects on the cardiovascular system than CPA. A concentration-dependent inhibition of ACh release was observed with a maximum of 60.5+/-2.4% for CPA, 42.5+/-2.3% for 2DCPA, 45.3+/-5.8% for 3DCPA, 57.1+/-1.4% for 8ECPA and 93.1+/-10.9% for 8BCPA, respectively. This effect was counteracted by the adenosine A(1) receptor antagonist 8-cyclopentyltheophylline. These findings show that low efficacy adenosine A(1) agonists inhibit striatal ACh release equally effective as CPA, suggesting that central nervous system-selective actions can be obtained with these compounds. PMID- 12749998 TI - Effects of ginseng saponin administered intraperitoneally on the hypothalamo pituitary-adrenal axis in mice. AB - Intraperitoneal injection of ginseng total saponin (GTS; 5 and 20 mg/kg) raised plasma corticosterone levels in mice. However, interestingly, pretreatment of animals with the same doses of GTS (5 and 20 mg/kg) significantly attenuated the immobilization stress-induced increase in plasma corticosterone levels. Of the ginsenosides Rb(1), Rb(2), Rc, Rd, Re, Rf, Rg(1), 20(S)-Rg(3), and 20(R)-Rg(3) injected intraperitoneally at doses of 0.1-2 mg/kg, Rc (2 mg/kg) significantly inhibited the immobilization stress-induced increase in plasma corticosterone levels. GTS and Rc administered intraperitoneally did not affect the immobilization stress-induced elevation of plasma adrenocorticotropic hormone (ACTH) level. Pretreatment with GTS and Rc significantly attenuated the increase in plasma corticosterone levels induced by intraperitoneal injection of ACTH (30 microg/kg). These results suggest that GTS and Rc inhibit the immobilization stress-induced increase in plasma corticosterone levels by blocking ACTH action in the adrenal gland. Ginseng may be proposed to be useful for treatment of stress related disorders. PMID- 12749999 TI - Axonal sprouting in mdx mice and its relevance to cell and gene mediated therapies for Duchenne muscular dystrophy. AB - We investigated whether pre-terminal axons and motor terminals retained their ability to sprout in the murine X-linked muscular dystrophy (mdx). Immunofluorescence confocal microscopy observation of nerve terminals and acetylcholine receptors in mdx muscles with crushed and non-crushed nerves showed that most of the junctions had intraterminal sprouting and that the number of junctions with extraterminal sprouting increased after the nerve crush lesion. Since new dystrophin-positive muscle fibers generated by cell-mediated therapies need to be innervated to proceed with their maturation and dystrophin production, these results suggest that the use of inducing factors to increase the sprouting capacity of nerve terminals could be an additional tool in the success of cell mediated therapies. PMID- 12750000 TI - No genetic association between polymorphisms in the Fyn kinase gene and age of schizophrenic onset. AB - Since the mRNA level of Fyn, a neurodevelopmental molecule, expression had been reported to be increased in postmortem schizophrenic prefrontal cortex and showed a strong correlation with age of disease onset, we investigated whether the three polymorphisms of the Fyn gene on genomic DNA (-93A/G, IVS10+37T/C and Ex12+894T/G) also had an effect on clinical onset in 139 unrelated schizophrenics. A comparison of the age of onset among the groups classified by polymorphisms showed no significant difference. Moreover, all allelic combinations also failed to show significant differences in age of onset among the groups. The present study reports that there is no indication that the three polymorphisms in the Fyn gene are associated with the age of schizophrenic onset. PMID- 12750001 TI - Induction of apoptosis by the p53-related p73 and partial inhibition by the baculovirus-encoded p35 in neuronal cells. AB - To better understand whether the p53-related p73 gene could induce neuronal apoptosis, we tested whether p73 induced cell killing in three neuronal cell lines and whether apoptosis could be inhibited by p35, a baculovirus-encoded protein that blocks caspase 3. Recombinant adenoviruses carrying the hemagglutinin (HA)-tagged p73beta or p35, or the green fluorescent protein gene driven by the cytomegalovirus immediate-early promoter were constructed, and used to infect human SK-N-AS and SK-N-SH neuroblastoma, and rat PC12 pheochromocytoma cells. Infection with Adp73beta virus resulted in p73beta over-expression and substantial reduction of cell viability due to apoptosis in all three neuronal cell lines as compared with the control AdGFP virus. These results indicate that p73beta over-expression in neuronal cells could induce apoptotic cell death regardless of the endogenous expression of p73. The p73 effect was partially blocked by co-expression of the wild-type p35, suggesting caspase-mediated cell killing. Insertion of a hemagglutinin (HA) tag at the N-terminus of p35 markedly reduced its ability to inhibit the p73 effect compared with the wild-type p35, while insertion of an HA tag to the C-terminus of p35 had no appreciable effect. Taken together, our results suggest that the N-terminal structure of p35 is critical for its anti-apoptotic activity on p73-induced apoptosis in neuronal cells. PMID- 12750002 TI - Identification of numerous genes differentially expressed in rat brain during postnatal development by suppression subtractive hybridization and expression analysis of the novel rat gene rMMS2. AB - During postnatal development the potential for axonal growth and regeneration in the central nervous system (CNS) becomes very restricted. This decline of axon growth and regeneration might be due to developmental alterations in the expression level of genes which are strongly expressed in differentiating neurons during formation of axons, but which are downregulated later in development. In order to identify genes which are downregulated in rat brain with the completion of neuronal differentiation, we performed suppression subtractive hybridization (SSH) with rat cerebellum at two developmental stages. Several differentially expressed genes were identified. We present the detailed expression analysis of one of these, rMMS2, which is the rat homologue of mouse ubiquitin-conjugating enzyme-like protein MMS2 and belongs to a family of ubiquitin-conjugating enzyme variants (UEVs) that are highly similar to ubiquitin-conjugating enzymes E2 (Ubcs) but lack the essential amino acid residue in the active site. UEVs play a role in DNA repair and are possibly involved in ubiquitination, which may be important for the assembly and function of neuronal circuits. In the present study, we examined the temporal and spatial expression of rMMS2 transcript and show a strong developmental downregulation in rat brain by Northern blot analysis and in situ hybridization. The mRNA of rMMS2 is widely distributed in rat brain at late embryonic development but is differentially regulated during postnatal development; its expression is strongly reduced during maturation of the CNS. Our results show that SSH is a suitable method for identifying genes which are regulated during postnatal development and suggest that the newly identified rat UEV rMMS2 may play a role in neuronal development and differentiation. PMID- 12750004 TI - Selective decrease in NR1 subunit splice variant mRNA in the hippocampus of Pb2+ exposed rats: implications for synaptic targeting and cell surface expression of NMDAR complexes. AB - We have previously shown that exposure to environmentally relevant levels of Pb(2+) during brain development decreases the expression of N-methyl-D-aspartate receptor (NMDAR) subunit 1 (NR1) and NR2A genes in the hippocampus of young adult rats and was associated with deficits in hippocampal LTP and spatial learning [Neuroscience 99 (2000) 233-242]. In the present study, we demonstrate that the lower levels of NR1 subunit mRNA expressed in the Pb(2+)-exposed hippocampus are principally due to decreased levels of the NR1-4 and NR1-2 splice variants. These changes were present in the absence of changes in GluR1, PSD-95 and alphaCaMKII gene expression. A unique characteristic of these splice variants is that they lack the C1 cassette. Further, these splice variants have been shown to impart the highest cell surface expression, PKC potentiation and calcium kinetics to NMDAR complexes. Our present findings indicate that Pb(2+)-induced changes in NR1 subunit splice variant mRNA expression in the hippocampus may provide a mechanism by which Pb(2+)-exposure can modify NMDAR-mediated calcium signaling and influence the degree of synaptic plasticity. PMID- 12750003 TI - Role of superoxide in poly(ADP-ribose) polymerase upregulation after transient cerebral ischemia. AB - Oxidative stress plays a pivotal role in ischemic-reperfusion cell injury. Oxygen derived free radicals trigger DNA strand damage, which is responsible for the activation of poly(ADP-ribose) polymerase (PARP). Recent studies have shown that peroxynitrite is the primary mediator of DNA damage and, hence, PARP activation after ischemia. PARP activation depletes NAD and ATP pools, ultimately resulting in necrotic cell death by loss of energy stores. Our study shows that PARP is upregulated as early as 15 min after 1 h of transient focal cerebral ischemia and remains for 8 h. We also examined the role of superoxide in PARP induction using copper/zinc-superoxide dismutase transgenic mice. Immunohistochemical and Western blotting data showed that there was no increased induction in PARP expression in these mice, suggesting that one of the mechanisms by which ischemic injury is attenuated in these mice might be by the inhibition of PARP induction. Furthermore, double staining of ischemic tissue with a PARP antibody and terminal deoxynucleotidyl transferase-mediated uridine 5'-triphosphate-biotin nick end labeling (TUNEL) indicated that most cells that are positive for TUNEL do not stain for the PARP antibody, confirming recent reports that PARP activation is involved in necrotic cell death rather than apoptosis during ischemic-reperfusion injury. PMID- 12750005 TI - Time- and cell type-specific induction of platelet-derived growth factor receptor beta during cerebral ischemia. AB - During cerebral ischemia, angiogenesis occurs inside and around the infarcted area. The growth of new blood vessels may contribute to a better outcome after stroke due to accelerated and increased delivery of nutrients and oxygen to the ischemic tissue. The platelet-derived growth factor (PDGF)-B/PDGF receptor (PDGFR)-beta system, hitherto thought to contribute mainly to neuroprotection, may also support angiogenesis and vascular remodeling by mediating interactions of endothelial cells with pericytes after cerebral ischemia. While platelet derived growth factor (PDGF)-B and its receptor PDGFR-beta are essential factors for the recruitment of pericytes to brain capillaries during embryonic development, their role in blood vessel maturation during cerebral ischemia is not clear. The aim of the present study was to investigate the time course and location of PDGF-B and PDGFR-beta expression in a mouse model of focal cerebral ischemia. In contrast to the early and continuous induction of PDGF-B, PDGFR-beta mRNA was specifically upregulated in vascular structures in the infarcted area 48 h after occlusion of the middle cerebral artery. Immunohistology and confocal microscopy analysis revealed the specific upregulation of PDGFR-beta on blood vessels and suggested expression mainly on pericytes. Our results imply PDGFR beta as a key factor in vascular remodeling during stroke and suggest that the pleiotropic functions of PDGF-B may be regulated via the expression of its receptor. Influencing the PDGF system therapeutically might improve angiogenesis, cellular protection, and edema inhibition. PMID- 12750006 TI - L-type high voltage-gated calcium channels cause an increase in diazepam binding inhibitor mRNA expression after sustained exposure to ethanol in mouse cerebral cortical neurons. AB - Mechanisms for increase in diazepam binding inhibitor (DBI) mRNA expression after sustained exposure to ethanol (EtOH) were investigated. Increases in 30 mM KCl induced [45Ca(2+)] influx and DBI mRNA expression after EtOH (50 mM) exposure for 3 days were completely abolished by nifedipine, but not by omega-agatoxin VIA and omega-conotoxin GIVA. These results indicate that EtOH-induced increase in DBI mRNA expression is mediated via increased Ca(2+) entry through up-regulated L type high voltage-gated calcium channels. PMID- 12750008 TI - Analysis of two D1-like dopamine receptors from the honey bee Apis mellifera reveals agonist-independent activity. AB - Dopamine is found in many invertebrate organisms, including insects, however, the mechanisms through which this amine operates remain unclear. We have expressed two dopamine receptors cloned from honey bee (AmDOP1 and AmDOP2) in insect cells (Spodoptera frugiperda), and compared their pharmacology directly using production of cAMP as a functional assay. In each assay, AmDOP1 receptors required lower concentrations of dopamine and 6,7-ADTN for maximal activation than AmDOP2 receptors. Conversely, butaclamol and cis(Z)-flupentixol were more potent at blocking the cAMP response mediated through AmDOP2 than AmDOP1 receptors. Expression of AmDOP1, but not AmDOP2, receptors significantly increased levels of cAMP even in the absence of ligand. This constitutive activity was blocked by cis(Z)-flupentixol. This work provides the first evidence of a constitutively activated dopamine receptor in invertebrates and suggests that although AmDOP1 and AmDOP2 share much less homology than their vertebrate counterparts, they display a number of functional parallels with the mammalian D1 like dopamine receptors. PMID- 12750007 TI - Ets transcription factors ER81 and Elk1 regulate the transcription of the human presenilin 1 gene promoter. AB - We have previously defined a crucial DNA element controlling 90% of the expression of the presenilin 1 gene at (-35 to +6). This region contains an Ets transcription factor binding motif, and a 2-base pair alteration within the core sequence (GGAA to TTAA) of the Ets consensus also reduced transcription by over 90%. We have shown that Ets1/2 transcription factors bind specifically to the -10 Ets element and activate PS1 transcription. The identification of other transcription factors recognizing specifically this promoter area should provide insights into the regulation of PS1. We have used the -10 Ets element as a bait in yeast one hybrid screening of a human brain cDNA library. This assay selected three factors from the Ets family: Ets2, ER81 and Elk1. We show that in vitro translated ER81 indeed binds specifically to the -10 region of the PS1 promoter and that ER81 activates by two- to threefold the basal transcription of a presenilin-1 promoter-chloramphenicol acetyltransferase reporter synthetic gene ( 119, +178)PS1CAT in transient infection assays in neuroblastoma cells (SK-N-SH). GABPalpha, a member of the Ets family closely related to Ets2 and also containing a pointed domain, only increased PS1 transcription by about twofold. Cotransfection of GABPbeta together with GABPalpha did not increase PS1 transcription. However, GABPbeta alone activated PS1 transcription by two- to threefold. In contrast, the more distantly related Ets factor Elk1 repressed PS1 transcription very effectively. PMID- 12750009 TI - Tumor necrosis factor-alpha triggers cell death of sensitized potassium chloride stimulated cholinergic neurons. AB - Cell death of cholinergic neurons of the basal forebrain plays an important role in neurodegenerative disorders, such as Alzheimer's disease. Inflammatory cytokines, such as, for example, tumor necrosis factor-alpha (TNF-alpha), may be involved in these neurodegenerative processes. The aim of this project was to study the role of TNF-alpha in the survival and nerve fiber growth of cholinergic neurons of the basal nucleus of Meynert in organotypic brain slices and in adult rats. Cholinergic neurons were visualized by immunohistochemistry for the enzyme choline acetyltransferase and nerve fibers by histochemistry for the enzyme acetylcholinesterase. When co-slices of basal nucleus of Meynert and neocortex were sensitized for 15 min with 30 mM potassium chloride and subsequently incubated for 1 week with 20 ng/ml TNF-alpha, cholinergic neurons and nerve fibers markedly degenerated. Incubation with different growth factors rescued the loss of cholinergic cell bodies and cholinergic nerve fibers. Injection of 30 mM potassium chloride and 50 ng TNF-alpha into four defined cortical regions of anesthetized adult rats resulted in predominant cell death of cholinergic neurons on the ipsilateral side. In conclusion, our data show that TNF-alpha potentiated cell death of cholinergic neurons possibly via retrograde axonal damage in vitro and in vivo. Cortical overactivation combined with an increased expression of pro inflammatory cytokines may contribute to the cell death observed in Alzheimer's disease and ageing. PMID- 12750010 TI - Selective modulation of lipopolysaccharide-stimulated cytokine expression and mitogen-activated protein kinase pathways by dibutyryl-cAMP in BV2 microglial cells. AB - Cyclic AMP is a very important regulator in a wide range of biological processes, including inflammatory reactions. To investigate the role of cAMP in microglia, we examined the effect of dibutyryl-cAMP (dbcAMP) on lipopolysaccharide (LPS) stimulated cytokine expression and signaling pathways in murine BV2 microglial cells. DbcAMP strongly suppressed LPS-induced TNF-alpha expression, without affecting NO, IL-6 or TGF-beta1 expression. In contrast, LPS-induced IL-1beta or IL-10 expressions were dramatically increased by dbcAMP. We further examined the effect of elevated cAMP on signaling molecules such as MAP kinases (p38 MAPK, ERK and JNK), NF-kappaB and AP1, which are involved in the regulation of inflammatory responses. DbcAMP decreased the LPS-induced phosphorylation of p38 MAPK, while it modestly enhanced the ERK activity. JNK phosphorylation was slightly reduced by dbcAMP only at the later time point. Electrophoretic mobility shift assay revealed that the elevated cAMP potentiated AP-1 binding activity by enhancing c fos binding. On the other hand, dbcAMP repressed NF-kappaB-mediated transcription without affecting NF-kappaB binding. Treatment with H89, a selective inhibitor of protein kinase A, completely reversed cAMP-induced IL-10 and IL-1beta upregulation but only partially reversed the cAMP-induced repression of TNF alpha. Thus, the effect of dbcAMP in BV2 cells appears to be mediated through both protein kinase A-dependent and -independent pathways. Taken together, our results demonstrate that cAMP modulates microglia activation in a diverse and complex manner. PMID- 12750012 TI - Altered expression of GABAB receptors in the hippocampus after kainic-acid induced seizures in rats. AB - Epilepsy is closely related to an altered transmission of GABA, the major inhibitory transmitter in the brain. GABA acts through two classes of receptors, ionotropic GABA(A) receptors and metabotropic GABA(B) receptors. Using in situ hybridization, receptor autoradiography and immunocytochemistry, we now investigated temporal changes in the expression the GABA(B)-1 and GABA(B)-2 subunits (GABA(B)-1R and GABA(B)-1R, respectively) in the hippocampus following kainic-acid-induced seizures. Significant decreases (by about 40%) in mRNA levels of both splice variants (a and b) of GABA(B)-1R and of GABA(B)-2R were observed in the principal cell layer of the hippocampus 6-12 h after kainic acid injection in the rat. Whereas mRNA levels in the granule cell layer returned to basal after 24 h, the decreases persisted in sectors CA1 and CA3, presumably due to progressing neurodegeneration. In the sector CA3, GABA(B)-R mRNA levels and GABA(B)-R1 immunoreactivity partially recovered 30 days after the initial kainic acid seizures indicating receptor upregulation in surviving neurons. PMID- 12750011 TI - Inwardly rectifying Kir3.1 subunit knockdown impairs learning and memory in an olfactory associative task in rat. AB - Inward-rectifier potassium channels gated by the direct action of G proteins are activated or inhibited by numerous neurotransmitters and they modulate neuronal excitability. Using an olfactory associative task, the effect of Kir3.1 subunit knockdown was tested on learning and memory. Repeated intracerebroventricular injections of antisense oligodeoxyribonucleotide to the Kir3.1 subunit significantly reduced hippocampal expression of its mRNA target determined by Western blotting. The antisense knockdown had no effect on locomotor and drinking activity or on attention processes. The reduction in Kir3.1 subunit impaired the learning of the odor associations and the procedural side of the task. This reduction correlated with the performance impairment. The results suggest that Kir3.1 channel activity is implicated in the memory processes. PMID- 12750013 TI - Regional specific increases of [3H]AMPA binding and mRNA expression of AMPA receptors in the brain of mu-opioid receptor knockout mice. AB - Previous pharmacological studies have indicated the possible existence of functional interactions between opioidergic and glutamatergic neurons in the CNS. In the present study, [(3)H]AMPA binding and the expression of mRNAs encoding flip and flop variants of three subtypes of AMPA glutamate receptor GluR1-3 were examined by in situ hybridization technique in order to investigate whether there is a change in the AMPA receptor system of mice lacking the mu-opioid receptor. In the mu-opioid receptor knockout mice, [(3)H]AMPA binding was increased in the hippocampal CA1 and dentate gyrus, cortex, and caudate putamen compared with that of the wild-type animals. The expression of GluR1 flip mRNA was increased in the cortex and caudate putamen of mu-opioid receptor knockout mice. The expression of GluR1 flop mRNA was increased in the cortex, caudate putamen, and hippocampal CA1 layer of mu-opioid receptor knockout mice. The expression of GluR2 flip mRNA was decreased in the hippocampal dentate gyrus of mu-opioid receptor knockout mice. The expression of GluR2 flop was not altered in any regions studied. The expression of GluR3 flip was increased in the cortical area and caudate putamen of mu-opioid receptor knockout mice. The expression of GluR3 flop was increased in the cortical area, hippocampal CA3 area, and caudate putamen of mu-opioid receptor knockout mice. These results indicate that [(3)H]AMPA binding and the expression of GluR1-3 mRNA were increased in a region and subunit specific manner, and suggest that changes in the AMPA receptor system are accompanied by the absence of mu-opioid receptor gene. PMID- 12750014 TI - Induction of galanin-like peptide gene expression in the rat posterior pituitary gland during endotoxin shock and adjuvant arthritis. AB - Galanin-like peptide (GALP) was recently isolated from the porcine hypothalamus. The GALP mRNA is restricted in the hypothalamic arcuate nucleus and pituicytes in the posterior pituitary grand (PP) of the rat. The physiological function of the GALP is not clear in both areas. We focused on the regulation of the GALP gene expression to determine the role of GALP in the PP. We examined the effects of potent stressors to modulate a pituitary function, nociceptive stimuli and acute and chronic inflammatory stresses on the expression of the GALP gene in the PP using in situ hybridization histochemistry. Subcutaneous (s.c.) injection of 5% formalin in the bilateral hind paws caused a moderate increase of GALP gene expression in the PP. Intraperitoneal (i.p.) injection of lipopolysaccharide (LPS) also caused a marked increase of GALP gene expression in the PP. Effects of i.p. injection of LPS on the expression of the GALP gene in the PP were attenuated by pretreatment with i.p. injection of indomethacin. The levels of GALP mRNA in the PP were markedly increased by two peaks at 24 h and 15 days after s.c. injection of heat-inactivated M. butyricum that induced adjuvant arthritis. These results suggested that inflammatory stresses may be a potent stimulant to induce the expression of the GALP gene in the PP. PMID- 12750015 TI - Fasting activates neuropeptide Y neurons in the arcuate nucleus and the paraventricular nucleus in the rhesus macaque. AB - It is well accepted that neuropeptide Y (NPY) plays a pivotal role in the regulation of food intake and energy homeostasis in the rodent, with NPY neurons in the arcuate nucleus (ARH) being thought of as the major contributor to the complex central feeding circuitry. Recent data from our group also indicate that NPY is important in the regulation of energy homeostasis in the nonhuman primate (NHP); exogenous NPY administration into the 3rd ventricle is a potent stimulator of food intake in the male rhesus macaque. The purpose of this study was to determine if NPY neurons in the rhesus macaque respond to a metabolic challenge, induced by 48 h of fasting, in a manner similar to that seen in the rodent. NPY mRNA was detected in hypothalamic sections from 48-h fasted or fed rhesus monkeys by in situ hybridization, using a [35S]UTP-labeled riboprobe specific for human NPY. Not surprisingly, NPY mRNA was abundant in the ARH of the NHP; however, of great interest was the expression of NPY mRNA in neurons within the paraventricular nucleus of the hypothalamus (PVH) and the supraoptic nucleus (SON). This raised the question as to whether all of these populations of NPY neurons are sensitive to changes in energy availability. Indeed, NPY expression in the ARH and PVH was significantly elevated in response to fasting; however, no significant change was detected in the SON. These data indicate that the NPY neurocircuitry involved in the regulation of food intake is more complex in the NHP than in rodents. PMID- 12750016 TI - Distribution of glucokinase, glucose transporter GLUT2, sulfonylurea receptor-1, glucagon-like peptide-1 receptor and neuropeptide Y messenger RNAs in rat brain by quantitative real time RT-PCR. AB - Glucokinase (GK), glucose transporter GLUT2, sulfonylurea receptor-1 (SUR1), glucagon-like peptide-1 receptor (GLP-1R) and neuropeptide Y (NPY) have been proposed to be involved in central glucose sensing or regulation of food intake. In this study, we combined tissue micropunch and real time reverse transcription polymerase chain reaction (RT-PCR), and measured GK, GLUT2, SUR1, GLP-1R and NPY mRNA expression in discrete areas in the hypothalamus and the hindbrain. PMID- 12750018 TI - Biotechnology and applied genomics for health: initiatives of the European Union. PMID- 12750019 TI - The design of potent, non-peptidic inhibitors of hepatitis C protease. AB - The pyrrolidine-5,5-trans-lactam template was used to design small, neutral, mechanism-based inhibitors of hepatitis C NS3/4A protease displaying potent activity in the replicon cell-based assay. The activity of this series is not dependent upon its chemical reactivity and molecules have been synthesised which combine enhanced biochemical potency with improved plasma stability. Promising initial pharmacokinetic data indicating the potential for further optimisation of this series into low molecular weight, drug-like inhibitors is presented. PMID- 12750020 TI - Inhibitors of phosphopantetheine adenylyltransferase. AB - Phosphopantetheine adenylyltransferase (PPAT) is an essential enzyme in Coenzyme A biosynthesis. Because bacterial PPAT and mammalian PPAT are dissimilar, this enzyme is an attractive antibacterial target. Based on the structure of the substrate, 4-phosphopantetheine, a dipeptide library was designed, synthesised and tested against Escherichia coli PPAT. The most potent inhibitor PTX040334 was co-crystallised with E. coli PPAT. With this structural information, a rational iterative medicinal chemistry program was initiated, aimed at increasing the number of inhibitor-enzyme interactions. A very potent and specific inhibitor, PTX042695, with an IC(50) of 6 nM against E.coli PPAT, but with no activity against porcine PPAT, was obtained. PMID- 12750021 TI - Lipopeptide substrates for SpsB, the Staphylococcus aureus type I signal peptidase: design, conformation and conversion to alpha-ketoamide inhibitors. AB - Pre-protein sequence data was used to design substrates for SpsB, the bacterial signal peptidase I enzyme from Staphylococcus aureus. Key elements were an alkyl membrane anchor, proline at P5 and lysine at P2. The proline at P5 induced a helical turn in the lipopeptide, as deduced from NMR studies, from P6 to P2 in membrane mimetic solvents. The substrate Decanoyl-LTPTAKAASKIDD-OH was cleaved by SpsB, as expected, between the P1 and P1' alanines with a k(cat)/K(m) of 2.3x10(6) M(-1)s(-1) at pH 8.5. Insertion of proline at P1' converted substrates to competitive inhibitors, whilst the incorporation of an alpha-ketoamide at the cleavage site transformed substrates to time dependent inhibitors of SpsB. PMID- 12750022 TI - Imidazolines as efficacious glucose-dependent stimulators of insulin secretion. AB - Synthesis of a series of imidazolines with glucose dependent effects on insulin exocytosis from pancreatic beta-cells is reported. Regioisomers and enantiomers were found to exhibit marked differences in exocytotic effects as well as different activities on the K(ATP)-channel; the (R (+)) isomer of 2-[2-(4,5 dihydro-1H-imidazol-2-yl)-1-thiophene-2-ylethyl]pyridine (4a) and the (+) isomer of 2-[2-(4,5-dihydro-1H-imidazol-2-yl)-1-thiophene-3-ylethyl]pyridine (4d) were found to give a significant increase in insulin release-in contrast to findings for their enantiomers-without influence on the K(ATP)-channel. The (+) isomer (4a) showed glucose dependent insulin release from beta-cells at concentrations above 2.5 mM and a marked glucose lowering effect in ob/ob mice as well as in fed but not in fasted rats. PMID- 12750023 TI - Discovery of selective CYP11B2 (aldosterone synthase) inhibitors for the therapy of congestive heart failure and myocardial fibrosis. AB - An increased aldosterone concentration due to congestive heart failure leads to a further progression of the disease as well as to myocardial fibrosis. To interfere with these fatal processes selective inhibition of aldosterone synthase (CYP11B2) is required. CYP11B1, a key enzyme in glucocorticoid biosynthesis showing a high homology to the target enzyme (>93%), must not be inhibited. Screening of our P450 inhibitor library for inhibition of bovine aldosterone synthase resulted in a high number of compounds showing reasonable inhibition. In the next step substances were tested for oral absorption using two artificial membrane assays. The inhibition of human CYP11B2 was evaluated using assays in fission yeast and V79MZ cells stably expressing the active human target enzyme. For selectivity, inhibition of CYP11B1, CYP11A1, CYP17, CYP19 and CYP5 was determined. Rather potent and selective compounds obtained in this way were structurally further optimised, finally leading to inhibitors showing IC(50) values within the low nanomolar range. PMID- 12750024 TI - Recent developments in the field of A2A and A3 adenosine receptor antagonists. AB - In the last years adenosine receptors have been extensively studied, and mainly at present we understand the importance of A(2A) and A(3) adenosine receptors. A(2A) selective adenosine receptors antagonists are promising new drugs for the treatment of Parkinson's disease, while A(3) selective adenosine receptors antagonists have been postulated as novel anti-inflammatory and antiallergic agents; recent studies also indicated a possible employment of these derivatives as antitumour agents. Lately different classes of compounds have been identified as potent A(2A) and A(3) antagonists. In this article we report the past and present efforts which led to development of more potent and selective A(2A) and A(3) antagonists. Our group has mainly worked on the pyrazolo[4,3-e]-1,2,4 triazolo[1,5-c]pyrimidine nucleus both as A(2A) and A(3) antagonists, aiming to improve the affinity, selectivity and the hydrophilic profile. In fact, we have synthesised several compounds endowed with high affinity and selectivity versus A(2A) adenosine receptors, as 2, 2a-c (K(i)A(2A)=0.12-0.19 nM), or A(3) adenosine receptors, as 4p (K(i)A(3)=0.01 nM) and 4q (K(i)A(3)=0.04 nM). PMID- 12750025 TI - Synthesis and HSA binding characterisation of the water soluble 7 succinylpaclitaxel. AB - A water soluble paclitaxel analogue, the 7-hemisuccinylpaclitaxel, was synthesised and its binding to human serum albumin (HSA) was characterised by difference circular dichroism and optical biosensor methodologies. The carboxylate group was introduced at paclitaxel C-7 position to improve the drug water solubility without significantly changing the biological activity. The paclitaxel analogue showed a relatively low affinity to HSA (3.5x10(4) M(-1)), while no significant interactions were evidenced with selective markers for the most characterised binding sites on the carrier, suggesting a non-selective binding to low affinity binding sites. PMID- 12750026 TI - Relative hydrophobicity and lipophilicity of drugs measured by aqueous two-phase partitioning, octanol-buffer partitioning and HPLC. A simple model for predicting blood-brain distribution. AB - Relative hydrophobicity and lipophilicity of 63 compounds with known permeability through the blood-brain barrier (BBB) was examined by partitioning in aqueous dextran-poly(ethylene glycol) two-phase system and octanol-buffer system, and by gradient RP-HPLC at pH 7.4. Combination of the relative hydrophobicity estimates, N(CH(2)) obtained by aqueous two-phase partitioning and the lipophilicity (logD(exp) or logD(HPLC)) values obtained by the shake-flask technique or HPLC technique allows one to differentiate between compounds capable of crossing the BBB and those that cannot. A simple model for predicting blood-brain distribution is proposed. PMID- 12750027 TI - Impact of the aryl substituent kind and distance from pyrimido[2,1-f]purindiones on the adenosine receptor selectivity and antagonistic properties. AB - Adenosine receptor (AR) antagonists belong to two major groups of compounds: xanthines and non-xanthines. Recently several annelated xanthine derivatives have been described as selective A(1), A(2A), A(2B) and A(3) ARs antagonists. Contrary to dipropyl derivatives, in the group of dimethyl (un)substituted arylalkyl pyrimido[2,1-f]purindiones selective mainly adenosine A(2A) receptor antagonists were identified. Their activity depended on aryl substitution and its distance from pyrimido[2,1-f]purindione. PMID- 12750028 TI - Design, synthesis and biological evaluation of new endocannabinoid transporter inhibitors. AB - In the present work we describe the synthesis and the in vitro evaluation of a series of arachidonic acid derivatives of general structure I as endocannabinoid transporter inhibitors. In addition, we report the first in vivo studies of the most potent derivative (4, UCM707) within this series. The majority of compounds studied are highly potent (IC(50)=24-0.8 micro M) and selective endocannabinoid uptake inhibitors with very low affinities for either the enzyme fatty acid amide hydrolase (IC(50)=30-113 micro M) or for cannabinoid receptor subtype 1 (CB(1)), cannabinoid receptor subtype 2 (CB(2)) and vanilloid receptor subtype 1 (VR(1)) (K(i)=1000-10000 nM). Among them, (5Z,8Z,11Z,14Z)-N-(fur-3-ylmethyl)icosa 5,8,11,14-tetraenamide (UCM707) behaves as the most potent endocannabinoid transporter inhibitor described to date (IC(50)=0.8 micro M) and exhibits improved potency for the anandamide transporter, high selectivity for CB(1) and VR(1) receptors, and modest selectivity for CB(2). In vivo it enhances the analgesia and hypokinetic effects induced by a subeffective dose of anandamide. PMID- 12750029 TI - Homology modelling and docking studies on Varicella Zoster Virus Thymidine kinase. AB - Thymidine kinase (TK) is the key enzyme in antiviral and suicide gene therapies. While herpes simplex virus type 1 thymidine kinase has been widely studied and crystallised less is known on Varicella Zoster Virus thymidine kinase (VZV TK) and its three-dimensional structure. In this paper we report the model of the three-dimensional structure of VZV TK resulting from a homology modelling study. Subsequent docking studies of the natural substrate deoxythymidine (dT) and known antiviral drugs were performed and shaded new light on the binding characteristics of the enzyme. PMID- 12750030 TI - A novel orally active inhibitor of HLE. AB - Human leukocyte elastase (HLE) is a serine proteinase, capable of degrading a variety of structural matrix proteins. SSR69071 2-[(4-isopropyl-6-methoxy-1,1 dioxido-3-oxo-1,2-benzisothiazol-2(3H)-yl)methoxy]-9-(2-piperidin-1-ylethoxy)-4H pyrido[1,2-a]pyrimidin-4-one was selected as a novel orally active HLE inhibitor for treatment of chronic obstructive pulmonary diseases, asthma, emphysema, cystic fibrosis and several inflammatory diseases (WO 01/44245 A1) (J. Pharm. Exp. Ther., submitted for publication). PMID- 12750031 TI - Prediction of oral bioavailability by adaptive fuzzy partitioning. AB - An adaptive fuzzy partition (AFP) algorithm was applied on two bioavailability data sets subdivided into four ranges of activity. A large set of molecular descriptors was tested and the most relevant parameters were selected with help of a procedure based on genetic algorithm concepts and stepwise method. After building several AFP models on a training set, the best ones were able to predict correctly 75% of the validation set compounds. Furthermore, an improvement of about 15% in the validation results was got, on the same data set, as regard to other prediction methods. The importance to work with data sets including a large molecular diversity, and to use tools able to manage it, was also shown. The prediction power was increased up to 25% employing a data set with a better optimised molecular diversity. PMID- 12750033 TI - Nitric oxide-donating non-steroidal anti-inflammatory drugs: the case of nitroderivatives of aspirin. AB - Nitric oxide (NO) acts as a key signalling mechanism in a number of cells and tissues in the mammalian organism. Modulation of the biosynthesis of NO has emerged to be relevant to the treatment of a variety of human diseases. In the attempt to reduce the serious side effects of non-steroidal anti-inflammatory drugs (NSAIDs), especially in the gastrointestinal tract, a NO-releasing moiety has been linked to conventional NSAIDs. A prototypical example is that of NO releasing derivatives of aspirin. Thanks to the cytoprotective action of NO such compounds do not produce gastric damage and are emerging as an interesting novel group of drugs for their unique pharmacological properties. PMID- 12750032 TI - Conformationally constrained butyrophenones as new pharmacological tools to study 5-HT 2A and 5-HT 2C receptor behaviours. AB - This study presents new pharmacological and molecular modelling studies on a recently described series of conformationally constrained butyrophenones. Alignment-free three-dimensional quantitative structure-activity relationship models developed on the basis of GRid Independent descriptors and partial least squares regression analysis, allow feasible predictions of activity of new compounds and reveal structural requirements for optimal affinity, particularly in the case of the 5-HT(2A) receptor. The requirements for the 5-HT(2A) affinity consist in a precise distance between hydrogen bond donor (protonated amino group) and hydrogen bond acceptor groups, as well as an optimal distance between the protonated amino group and the farthest extreme of the compounds. Another significant result has been the characterisation of two structurally similar compounds as interesting pharmacological tools (1-[(4-Oxo-4,5,6,7 tetrahydrobenzo[b]furan-5-yl)ethyl]-4-(6-fluorobenzisoxazol-3-yl)piperidine and 1 [(4-Oxo-4,5,6,7-tetrahydrobenzo[b]furan-6-yl)methyl]-4-(6-fluorobenzisoxazol-3 yl)piperidine). In spite of their structural similarity, the first compound shows clearly higher affinity for the 5-HT(2C) receptor (about 100 fold) and higher Meltzer ratio (1.17 vs. 0.99) than the second. Moreover, the first compound inhibits arachidonic acid release in a biphasic concentration-dependent way in functional experiments at the 5-HT(2A) receptor and it acts as inverse agonist at the 5-HT(2C) receptor, behaviours that are not shown by the second compound. PMID- 12750034 TI - Design and synthesis of a new series of 4-alkylated 3-isoxazolol GABA A antagonists. AB - A number of analogues of the low-efficacy partial GABA(A) agonist 5-(4-piperidyl) 3-isoxazolol (4-PIOL), in which the 4-position of the 3-isoxazolol ring is substituted by different groups, were synthesized and tested as GABA(A) receptor ligands. While alkyl and benzyl substitution provided affinities and antagonist potencies comparable to those of 4-PIOL, diphenylalkyl and naphthylalkyl substitution resulted in marked increase in both affinity and potency. The 2 naphthylmethyl and the 3,3-diphenylpropyl analogues showed antagonist potencies comparable or markedly higher than that of the standard antagonist SR 95531. Molecular modeling studies exposed a large cavity in the vicinity of the 4 position of 4-PIOL, in which there seems to be additional sites for specific receptor interactions. PMID- 12750035 TI - Focused section on QT correction for heart rate. PMID- 12750036 TI - How to correct the QT interval for the effects of heart rate in clinical studies. AB - Much inter- and intra-subject variability in the QT interval in health and disease is accounted for by differences in heart rate, leading to difficulties when determining the effects of disease and drugs on the QT interval. Traditionally, heart rate correction formulae have been used to overcome this problem in man. However, the commonly used Bazett's heart rate correction formulae (QT=QT(C) radical RR interval) does not remove the effect of heart rate; indeed, it overcorrects at high heart rates. Fredericia's formula (QT=QT(C)x(3) radical RR interval) does remove the effects of heart rate; this is the preferable formula, if one is to be used. However, all formulae make assumptions about the nature of the QT-heart rate relationship, assumptions that may not apply to those with disease or on drugs. A more intellectually rigorous approach to QT interval-heart rate correction is to determine the QT-heart relationship for each individual, using data obtained from exercise tests or 24-h Holter tapes. The best mathematical relationship (linear, exponential, etc.) is obtained from analysis of this data, and is used to determine the QT interval at a heart rate of 60 bpm, the QT(60). The QT(60) measure makes no assumptions about the nature of the QT interval-heart rate relationship, removes the dependence of QT interval on heart rate, and maintains genuine biological differences in the QT interval. It should become the standard in QT interval-heart rate correction. PMID- 12750037 TI - A method for QT correction based on beat-to-beat analysis of the QT/RR interval relationship in conscious telemetred beagle dogs. AB - INTRODUCTION: Drug-induced QT prolongation is a major clinical risk factor for arrhythmia induction, particularly torsades de pointes. QT interval is rate dependent, and many formulae exist that attempt to correct QT for changes in heart rate. Most correction factors are acknowledged to overcorrect at high heart rates, undercorrect at low heart rates, and tend to be species specific. Data collected from computerised data acquisition systems are normally reported as means over a given logging period, and so extremes of heart rate are averaged out. Therefore, the aim of this study was to develop a technique for assessing drug-induced changes in the QT/RR relationship, which is simple, suitable for small group sizes, and better able to determine rate-dependent effects of drugs. METHODS: Telemetred beagle dogs (n=4) instrumented for the measurement of electrocardiogram (ECG) were monitored for four separate 20-h periods to define the control QT/RR relationship. Data were binned by RR interval, in 10 ms bins, to produce a control curve. Each dog was treated with vehicle and sotalol (4, 8, 32 mg/kg) in a crossover design to determine whether drug-induced changes in the QT/RR relationship could be detected using the data binning technique. RESULTS: The control QT/RR relationship was curvilinear with a steep section for RR intervals below 580 ms, and was much less steep after this point. Sotalol produced QT prolongation and bradycardia-Fridericia's correction (QTf) reduced the magnitude of this prolongation. The data analysed by the binning technique showed a larger prolongation in QT than was suggested by QTf, and an inverse frequency-dependent response. DISCUSSION: Beat-to-beat analysis and binning allows accurate determination of the QT/RR relationship and assessment of QT prolongation without recourse to mathematical modelling. It also highlights the importance of assessing QT effects in well-trained animals over a range of heart rates. PMID- 12750038 TI - A new model for the continuous monitoring of polymorphonuclear leukocyte trapping in the pulmonary vasculature of the rabbit. AB - INTRODUCTION: Accumulation of polymorphonuclear leukocytes (PMNs) within the pulmonary vasculature contributes to the pathophysiology of a number of diseases, including chronic obstructive pulmonary disease (COPD) and acute respiratory distress syndrome. The techniques available to study this have their limitations. METHODS: We have developed a minimally invasive technique for the continuous monitoring of 111In-labelled PMNs (111In-PMNs) in the thoracic and groin regions of rabbits. The effects of intravenous injection of the chemoattractants, interleukin (IL)-8, leukotriene (LT) B(4), and lipopolysaccharide (LPS) were studied. RESULTS: Intravenous injection of LTB(4), IL-8, or LPS caused an accumulation of 111In-PMNs in the lung and a concomitant decrease in counts in the leg and caused no significant change from baseline in counts in the lung or the leg after injection of 111In-labelled red blood cells (RBCs). CONCLUSION: In conclusion, we have demonstrated a technique that is simple, reproducible, and robust, which can be used to investigate PMN trapping in the lung vasculature in response to well characterised PMN chemoattractants and that may be applied to the preclinical study of new drugs. PMID- 12750039 TI - A new screening model for safety evaluation of superantigen-antibody recombinant fusion proteins (mAb Fab-SEA/E) using telemetric monitoring in conscious rabbits. AB - INTRODUCTION: Carcinoma recognising monoclonal antibodies (mAb) and mutated forms of the T-cell-activating bacterial staphylococcal enterotoxin A/E (SEA/E) have been combined in single hybrid constructs (mAb Fab-SEA/E). By introducing substitutions in an MHC class II binding site, these harmful toxins can be converted into tolerable immunotoxins. Rabbits and humans are sensitive to SE toxins, and cardiovascular effects in rabbits are similar to those seen in septic shock in man. A new screening model using telemetry in conscious rabbits was applied in the safety evaluation of different mAb Fab-SEA/E constructs administered intravenously. METHODS: Telemetry transmitters were implanted in the peritoneal cavity of animals with a pressure catheter in the aorta and electrodes for ECG recording subcutaneously following administration of mAb Fab-SEA/E constructs intravenously. RESULTS: The responses in body temperature, heart rate, and blood pressure varied depending on the treatment regimen and the mutations of the drug given. For example, 25 micro g/kg of C215 Fab-SEAmut9 were given as a first treatment cycle on days 1, 5, and 7 and as a second treatment cycle on days 13-15. The first dose induced high fever, whereas the second and third doses induced fever responses more rapidly and were of lower and shorter duration. The second treatment cycle, starting on day 13, did not induce any responses probably due to anti-SEA antibodies formed because of the treatment. Another construct, 5T4 Fab-SEA/E-11 at 50 micro g/kg, induced a similar response as C215 Fab-SEAmut9 on days 1, 5, and 7. In this case, the pharmacologic response was still present on days 13-15, though no clinical signs developed or no formation of anti-SEA antibodies occurred. When 50 micro g/kg of 5T4 Fab-SEA/E-11 was administered once daily for 4 days, body temperature after the first dose increased slowly during the first 24 h, whereas the second to fourth doses induced more rapid and higher responses. The fourth dose of another compound, K305 Fab-SEA/E-11 (50 micro g/kg), induced an even more pronounced response both in magnitude and in duration as well as in adverse clinical signs. DISCUSSION: By using continuous telemetric registration in the rabbit as a tool in superantigen-antibody (mAb Fab-SEA/E) drug selection, it has been possible to evaluate the dynamics of drug-induced immune effects (fever) and concomitant engagement of the cardiovascular system, conditions that are essential before clinical trials can be initiated. PMID- 12750040 TI - Optimization of the dose and route of injection, and characterisation of the time course of carbon tetrachloride-induced hepatotoxicity in the rat. AB - INTRODUCTION: The purpose of this study was to optimize carbon tetrachloride induced hepatotoxicity in the rat with respect to dose, route of injection, and time course. METHODS: Male Wistar albino rats, 4 to 6 weeks old and weighing 130 180 g were used. Hepatotoxicity was evaluated by measuring the activity of serum enzymes (alkaline phosphatase [ALP], alanine aminotransferase [ALT], and aspartate aminotransferase [AST]) as well as serum total bilirubin level. RESULTS: Intraperitoneal injection of carbon tetrachloride (CCl(4)) increased the activity of ALP (from 64.9 to 137.3 U/l), ALT (from 106.6 to 693.1 U/l), and AST (from 113.8 to 693.9 U/l). Plasma bilirubin level increased (from 0.119 to 0.42 mg/dl). In contrast, subcutaneous injection of CCl(4) had no effect on these variables. The optimum intraperitoneal dose of CCl(4) was found to be 2 ml/kg body weight (dissolved in an equal volume of olive oil), and this increased the level of bilirubin and the activity of the three enzymes significantly, without causing death of the animals. Hepatotoxicity was observed within 2 h of intraperitoneal injection of CCl(4) and reached a peak after 24 h. Bilirubin level and serum enzyme activities declined gradually to normal levels by 3 days after CCl(4) injection. CONCLUSION: It is possible to reliably evoke reversible hepatotoxicity in rats by intraperitoneal injection of 2 ml/kg CCl(4). PMID- 12750041 TI - Use of the jaw opening reflex for assessing the effects of local anaesthetics in freely moving rats. AB - INTRODUCTION: In order to characterize a nonbehavioral model for assessing local anaesthetic (LA) activity, the effects of different LA agents (articaine, bupivacaine, procaine, and tetracaine) were measured in the conscious rat using the jaw-opening reflex (JOR). METHODS: One hundred sixty rats were chronically implanted with stimulating electrodes in the dental pulp of the low incisor. While animals were conscious and unrestrained, the JOR threshold was measured electrophysiologically via electrodes wrapped around the digastric muscle. Each LA was administered in the infratemporal area. The increase of the JOR threshold was assessed during a 3-h period following injection. RESULTS: Statistical analysis of the data showed a dose-dependent response to the four drugs tested. When the highest dose of each drug (articaine and procaine: 24 mg kg(-1), bupivacaine: 6 mg kg(-1), tetracaine: 3 mg kg(-1)) was administered (i) an immediate effect was observed for tetracaine and bupivacaine, whereas a 5-min delay was needed for articaine and procaine to act on the JOR threshold and (ii) an increase (>60%) of the JOR threshold was observed. The effects lasted 90 min for articaine, 45 min for procaine and bupivacaine, and 15 min for tetracaine before a return to baseline values. DISCUSSION: The rat JOR response combined with infratemporal injection of test drugs can be used for the pharmacological evaluation of LAs. PMID- 12750044 TI - Multiple study trials and judgments of learning. AB - We compared judgments of learning (JOLs) that were made either (a) after 1 study trial, (b) 2 study trials, or (c) in-between the 1st and 2nd study trials. In regard to the absolute accuracy of JOLs at predicting subsequent recall, we replicated previous findings of an underconfidence-with-practice effect for immediate JOLs and report for the first time a new finding of an underconfidence with-practice effect for delayed JOLs (i.e., delayed JOLs after one trial overestimated the likelihood of subsequent recall, whereas delayed JOLs after two trials underestimated that likelihood). Also, although delayed JOLs always had a greater relative accuracy than did immediate JOLs, the relative accuracy of immediate and delayed JOLs was approximately the same after 1 versus 2 study trials. These results demonstrate that additional study trials affect the absolute accuracy of all JOLs but not the relative accuracy of any JOLs. Thus an increase in the number of study trials produced an increasing bias to be underconfident about the subsequent likelihood of recall but did not affect people's ordering of which items had been more (versus less) well-learned. PMID- 12750042 TI - A nonradioisotopic endpoint for measurement of lymph node cell proliferation in a murine allergic contact dermatitis model, using bromodeoxyuridine immunohistochemistry. AB - INTRODUCTION: The murine local lymph node assay (LLNA) was developed as an alternative to guinea pig models for the assessment of the xenobiotic contact sensitization potential. However, it would be advantageous to have an alternative endpoint to the usual radioisotopic-dependent measures. In the present study, we investigated the development of a nonradioisotopic endpoint for LLNA using immunohistochemistry. METHODS: Female Balb/c mice were treated by the topical application of strong sensitizers, 2,4-dinitrochlorobenzene (DNCB) and toluene diisocyanate (TDI), and a strong irritant, sodium lauryl sulfate (SLS), on the dorsum of both ears once daily for three consecutive days. The proliferation of cells in the auricular lymph node and ears was analyzed by means of the labeling index (LI) of bromodeoxyuridine (BrdU) incorporation into cells. RESULTS: Skin reactions, consisting of increased ear thickness and the presence of inflammatory cell infiltrates, were observed in mice treated with DNCB and TDI. The cell number and the weight of the lymph nodes in the mice treated with the allergens, DNCB and TDI, were increased compared to vehicle control. We observed an increase in the areas of the B220(+) cells in the lymph nodes of mice treated with allergens, as determined by immunohistochemistry. There was an increase in the percentage of B220(+) cells in mice treated with DNCB and TDI compared to the vehicle control, but not in those treated with SLS. Because we observed an increase in the percentage of B cells in the allergen-treated group, we measured the stimulation index (SI) in the cortex and medulla (C+M) of the lymph node. The SI values of the C+M in the lymph nodes of the mice treated with DNCB and TDI were increased more than threefold compared with that of the control. However, the SI of the C+M in the lymph nodes of the mice exposed to 25% SLS was not significantly increased compared to the vehicle control, although the lymph node weight of the SLS group was significantly increased. DISCUSSION: In Balb/c mice, BrdU immunohistochemistry showed its potential use for the identification and differentiation of chemicals with the capacity to induce irritation and sensitization. The results suggest that the measurement of the SI in the cortex and medulla of the lymph node using BrdU immunohistochemistry could provide a useful method to screen irritants and allergens. PMID- 12750045 TI - Attentional capture within and between objects. AB - The present study addressed the question whether attentional capture by abrupt onsets is affected by object-like properties of the stimulus field. Observers searched for a target circle at one of four ends of two solid rectangles. In the focused attention condition the location of the upcoming target was cued by means of a central arrowhead, whereas in the divided attention condition, the target location was not cued. Irrelevant abrupt onsets could appear either within the attended or within the non-attended object. The results showed that in the focused attention condition, onsets ceased to capture attention irrespective of whether the onset appeared within an attended object or within a non-attended object. PMID- 12750046 TI - Lexical, conceptual and motor information in memory for action phrases: a multi system account. AB - According to the multi-system account [J. Engelkamp, H.D. Zimmer, The Human Memory: A Multi-Modal Approach, Hogrefe & Huber, Seattle, 1994], the enactment effect in free recall of action phrases (e.g. break the stick) is independent of inter- and intra-phrase associations because it emerges from the nonverbal encoding processes under enactment. Two experiments have been reported which tested these and further assumptions of the multi-system account of the enactment effect. In both experiments, inter- and intra-phrase (verb-object) associations were varied simultaneously in addition to enactment. In Experiment 1, the memory test was free recall, and in Experiment 2, it was cued recall. Independent effects of all three factors were observed in free recall. Enacted phrases were recalled better than phrases learned only verbally. Phrases with high-associated objects and verbs were better recalled than phrases with low-associated objects and verbs, and categorically related phrases were better recalled than unrelated phrases. In cued recall, there was no effect of categorical list structure. The effects of intra-phrase structure and enactment corresponded to those in free recall. All interactions were nonsignificant. The findings were interpreted as support for the multi-system account. PMID- 12750047 TI - Syllabic length effects in visual word recognition and naming. AB - Two experiments investigated the role of the number of syllables in visual word recognition and naming. Experiment 1 (word and nonword naming) showed that effects of number of syllables on naming latencies were observed for nonwords and very low-frequency words but not for high-frequency words. In Experiment 2 (lexical decision), syllabic length effects were also obtained for very low frequency words but not for high-frequency words and nonwords. These results suggest that visual word recognition and naming do require syllabic decomposition, at least for very low-frequency words in French. These data are compatible with the multiple-trace memory model for polysyllabic word reading [Psychol. Rev. 105 (1998) 678]. In this model, reading depends on the activity of two procedures: (1) a global procedure that operates in parallel across a letter string (and does not generate a strong syllabic length effect) and that is the predominant process in generating responses to high-frequency words, and (2) an analytic procedure that operates serially across a letter string (and generates a strong syllabic length effect) and that is the predominant process in generating responses to very low-frequency words. A modified version of the dual route cascaded model [Psychol. Rev. 108 (1) (2001) 204] can also explain the present results, provided that syllabic units are included in this model. However, the Parallel Distributed Processing model [Psychol. Rev. 96 (1989) 523; J. Exp. Psychol.: Human Perception Perform. 16 (1990) 92] has difficulties to account for these results. PMID- 12750048 TI - Temporal parameters and time course of perceptual latency priming. AB - Visual stimuli (primes) reduce the perceptual latency of a target appearing at the same location (perceptual latency priming, PLP). Three experiments assessed the time course of PLP by masked and, in Experiment 3, unmasked primes. Experiments 1 and 2 investigated the temporal parameters that determine the size of priming. Stimulus onset asynchrony was found to exert the main influence accompanied by a small effect of prime duration. Experiment 3 used a large range of priming onset asynchronies. We suggest to explain PLP by the Asynchronous Updating Model which relates it to the asynchrony of 2 central coding processes, preattentive coding of basic visual features and attentional orienting as a prerequisite for perceptual judgments and conscious perception. PMID- 12750049 TI - Age-related effects of attentional and oculomotor capture by onsets and color singletons as a function of experience. AB - The present experiment examined the degree to which experience with different stimulus characteristics affects attentional capture, particularly as related to aging. Participants were presented with onset target/color singleton distractor or color singleton target/onset distractor pairs across three experimental sessions. The target/distractor pairs were reversed in the second session such that the target in the first session became the distractor in the second and third sessions. For both young and old adults previous experience with color as a target defining feature influenced oculomotor capture with task-irrelevant color distractors. Experience with sudden onsets had the same effect for younger and older adults, although capture effects were substantially larger for onset than for color distractors. Experience-based capture effects diminished relatively rapidly after target and distractor-defining properties were reversed. The results are discussed in terms of top-down and stimulus-driven effects on age related differences in attentional control. PMID- 12750051 TI - Three-dimensional lamina architecture alters light-harvesting efficiency in Fagus: a leaf-scale analysis. AB - Modification of foliage exposition and morphology by seasonal average integrated quantum flux density (Qint) was investigated in the canopies of the shade tolerant late-successional deciduous tree species Fagus orientalis Lipsky and Fagus sylvatica L. Because the leaves were not entirely flat anywhere in the canopy, the leaf lamina was considered to be three-dimensional and characterized by the cross-sectional angle between the leaf halves (theta). Both branch and lamina inclination angles with respect to the horizontal scaled positively with irradiance in the canopy, allowing light to penetrate to deeper canopy horizons. Lamina cross-sectional angle varied from 170 degrees in the most shaded leaves to 90-100 degrees in leaves in the top of the canopy. Thus, the degree of leaf rolling increased with increasing Qint, further reducing the light-interception efficiency of the upper-canopy leaves. Simulations of the dependence of foliage light-interception efficiency on theta demonstrated that decreases in theta primarily reduce the interception efficiency of direct irradiance, but that diffuse irradiance was equally efficiently intercepted over the entire range of theta values in our study. Despite strong alteration in foliage light-harvesting capacity within the canopy and greater transmittance of the upper crown compared with the lower canopy, mean incident irradiances varied more than 20-fold within the canopy, indicating inherent limitations in light partitioning within the canopy. This extensive canopy light gradient was paralleled by plastic changes in foliar structure and chemistry. Leaf dry mass per unit area varied 3-4-fold between the canopy top and bottom, providing an important means of scaling foliage nitrogen contents and photosynthetic capacity per unit area with Qint. Although leaf structure versus light relationships were qualitatively similar in all cases, there were important tree-to-tree and species-to-species variations, as well as evidence of differences in investments in structural compounds within the leaf lamina, possibly in response to contrasting leaf water availability in different trees. PMID- 12750052 TI - Sources of variation in ecophysiological parameters in Douglas-fir and grand fir canopies. AB - Forest process models predict ecosystem responses from climate variables and physiological parameters. The parameters describe key ecosystem attributes, often as lumped averages. However, the sources and magnitude of variation in these physiological parameters are unknown, which complicates sampling if models are to be parameterized with field measurements. We measured several key parameters, which had been identified by sensitivity analyses of three models, in Abies grandis (Dougl.) Lindl. and Pseudotsuga menziesii var. glauca (Beissn.) Franco trees throughout the growing season. Trees were sampled at eight sites across the interior northwest of the USA. At each site, fertilized and control plots were sampled. The design provided statistical replication for the analysis of variance within a site, allowing us to draw inferences about a regional population of stands. Specific leaf area (SLA) varied by canopy position and treatment (P = 0.0003), by date of sampling (P < 0.0001) and by species (P = 0.0188). Mass-based foliar nitrogen concentration (%N) increased during the summer in both species (P = 0.0019), but at a faster rate in P. menziesii var. glauca than in A. grandis. Sun foliage had a higher mean %N (1.00, SE = 0.02%) than shade foliage (0.92 +/- 0.01%). Apparent quantum yield (Phi) varied among treatments, between species and between canopy positions; each of these variables interacted with date of sampling (P = 0.0207, P < 0.0001 and P = 0.0344, respectively). In A. grandis, mean Phi values (+/- SE) were 0.048 +/- 0.006 mol CO2 (mol incident photon)-1 for sun foliage and 0.067 +/- 0.007 mol CO2 (mol incident photon)(-1) for shade foliage. In P. menziesii, the corresponding mean Phi values were 0.032 +/- 0.003 and 0.047 +/- 0.004 mol CO2 (mol incident photon)-1. Parameters SLA, %N and Phi were all influenced by date, fertilizer treatment, species and crown position. We discuss methods of inferring quantum yields from light response curves and their utility for parameterizing process models. Parameter mean values are presented for each site; these tables provide a documented data set for the parameterization of models describing the western interior forests of the USA. PMID- 12750053 TI - Physiological responses of birch (Betula pendula) to ozone: a comparison between open-soil-grown trees exposed for six growing seasons and potted seedlings exposed for one season. AB - Physiological responses of 4-year-old potted saplings of an O3-tolerant clone of Betula pendula Roth to short-term ozone (O3) exposure (one growing season) were compared with those of 6-year-old open-soil-grown trees of the same clone fumigated with O3 for six growing seasons. In the 2001 growing season, both groups of plants were exposed to ambient (control) and 1.6x ambient (elevated) O3 concentration under similar microclimatic conditions in a free air O3 exposure facility. Growth, net photosynthesis, stomatal conductance, stomatal density, visible foliar injury, starch and nutrient concentrations, bud formation and differences in O3 responses between lower, middle and upper sections of the canopy were determined. The potted saplings were unaffected by elevated O3 concentration, whereas the open-soil-grown trees showed a 3-38% reduction in shoot growth, a 22% reduction in number of overwintering buds, a 26-65% decrease in autumnal net photosynthesis, 30% and 20-23% reductions in starch and nitrogen concentrations of senescing leaves, respectively, and disturbances in stomatal conductance. The greater O3 sensitivity of open-soil-grown trees compared with potted saplings was a result of senescence-related physiological factors. First, a lower net photosynthesis to stomatal conductance ratio in open-soil-grown trees at the end of the season promoted O3 uptake and decreased photosynthetic gain, leading to the onset of visible foliar injuries. Second, decreased carbohydrate reserves may have resulted in deleterious carry-over effects arising from the reduced formation of over-wintering buds. Finally, the leaf-level O3 load was higher for open-soil-grown trees than for potted saplings because of slower leaf senescence in the trees. Thus, O3 sensitivity in European white birch increases with increasing exposure time and tree size. PMID- 12750054 TI - Effects of simulated thaw on xylem cavitation, residual embolism, spring dieback and shoot growth in yellow birch. AB - Yellow birch seedlings (Betula alleghaniensis Britt.) that had lost more than 90% of their stem hydraulic conductivity during ambient winter temperatures were exposed to 0 and 20 days of a simulated winter thaw followed by a 48-h freezing treatment at 0, -5, -10, -20 and -30 degrees C. After measuring freezing injury to shoots and roots, the seedlings were placed in a greenhouse where recovery of xylem conductivity and new growth were measured. Shoot xylem cavitation was measured as percent loss of hydraulic conductivity. Shoot freezing injury was assessed by electrolyte leakage (EL) and root freezing injury was assessed by EL and triphenyl tetrazolium chloride reduction. Seedlings pretreated with thaw had higher stem water contents and suffered more freezing damage to roots and shoots (at -20 and -30 degrees C, respectively) than unthawed seedlings. After 3 weeks in a greenhouse, seedlings from the 0, -5 and -10 degrees C freezing treatments showed complete recovery of xylem conductivity, with substantially increased stem water contents. Poor recovery of hydraulic conductivity was observed only in seedlings that were subjected to freezing treatments at -20 and -30 degrees C, regardless of thaw treatment. Of these embolized seedlings, however, only those not previously thawed showed recovery of hydraulic conductivity or regained stem water content after 9 weeks in the greenhouse. Shoot dieback, bud burst and length of new shoots were significantly related to the extent of stem xylem cavitation and freezing injury. We conclude that (1) the simulated winter thaw predisposed yellow birch seedlings to freezing damage in shoots and roots by dehardening tissues and increasing their water content; (2) root freezing damage in turn affected the seedlings' ability to refill embolized stem xylem, resulting in considerable residual xylem embolism after spring refilling; (3) further recovery of stem xylem conductivity was attributable to growth of new vessels; (4) and the permanent residual embolism, together with root and shoot freezing injury, caused increased dieback, bud mortality and reduced growth of new shoots. PMID- 12750055 TI - Selection of a seed orchard of Eucalyptus dunnii based on genetic diversity criteria calculated using molecular markers. AB - A Eucalyptus dunnii Maiden breeding population of 46 accessions originated in Australia and selected for fitness to subtropical and cold environments was screened by Amplified Fragment Length Polymorphism (AFLP) and microsatellite markers to obtain quantitative estimates of genetic diversity. A randomly chosen group of AFLP primers generated 205 AFLP bands that were used to fingerprint the genotypes and to evaluate genetic relationships among accessions. Sixty-eight percent (140) of the bands were polymorphic markers. The mean diversity index (DI) was 0.33 and about 52% of the loci had values greater than 0.4. Cluster analysis derived from similarity indices (SI) revealed no particular grouping among accessions suggesting the absence of closely related genotypes, except for five pairs of genotypes. Bootstrap analysis results confirmed the suitability of AFLP to describe genetic relationships in this breeding population. In addition, four highly informative microsatellites were used to construct an identification matrix that discriminated nearly all of the genotypes. Mean values for the number of alleles per locus, DI and SI among accessions were 13, 0.78 and 0.19, respectively, indicating that the breeding population has high genetic diversity. However, several genotypes showed the presence of single microsatellite bands suggesting a putatively important degree of homozygosity. Molecular data were used to design a clonal seed orchard. To achieve this aim, the nine most divergent pairs of genotypes were chosen, thereby retaining 95.2% of the total number of alleles from the 140 polymorphic AFLP loci and the four microsatellite loci analyzed. Mean DI and SI for AFLP and microsatellites showed no significant differences between the original breeding population and the selected seed orchard, confirming that a seed orchard can be designed with a limited number of individuals, which allows similar accessions to be discarded and avoids inbreeding. PMID- 12750056 TI - Isolation and characterization of a cDNA from Quercus robur differentially expressed in juvenile-like and mature shoots. AB - A full-length cDNA clone named QRCPE (Quercus robur crown preferentially expressed) that is differentially expressed during in vitro culture of mature and juvenile-like shoots of Quercus robur L. was identified by differential display. The deduced amino acid sequence showed that the encoded protein is small, contains a predicted N-terminal hydrophobic signal peptide that targets the protein to the cell wall, and is rich in glycine and histidine residues. Accumulation of QRCPE mRNA was higher in oak microshoots derived from crown branch shoot cultures than in oak microshoots derived from basal shoot cultures at the end of the multiplication and rooting period. Among organs, the highest accumulation of QRCPE transcripts was detected in roots, followed by stems and leaves, with preferential accumulation in specific organs of ontogenetically older shoots. Although QRCPE mRNA was abundant in oak zygotic and somatic embryos, almost no QRCPE mRNA accumulation was detected in nodular callus cells, suggesting a possible role of this gene in embryonic development. In proliferating shoot cultures of two chestnut (Castanea sativa Mill.) clones, the QRCPE homolog was preferentially expressed in crown-derived shoots. On the other hand, expression analysis of QRCPE in juvenile and mature material from soil grown oak plants indicated that this gene is expressed from the embryonic to mature phases, but is progressively down-regulated during plant maturation. In vitro culture conditions induced changes in QRCPE transcript abundance in both basal and crown shoots in a phase-dependent manner. We conclude that QRCPE expression in oak is correlated with the ontogenetic stage of shoots, and thus this gene may be useful as a potential molecular marker for maturation-related characteristics. PMID- 12750057 TI - Explanation of vegetation succession in subtropical southern China based on ecophysiological characteristics of plant species. AB - A stomatal conductance model and a photosynthesis model were applied to field measurements of transpiration and photosynthesis of seven tree species growing in subtropical southern China. Parameter values of drought resistance and tolerance and biochemical assimilation capacity were obtained by means of nonlinear statistical regression, and were used to quantify species succession. The analysis indicated that the models adequately described the ecophysiological behavior of the trees under various environmental conditions. We found a general pattern of decreased drought resistance and tolerance, but increased biochemical assimilation capacity from pines to heliophilus broadleaf trees to mesophilus broadleaf trees. Succession was explained on the basis of these physiological characteristics together with positive feedbacks caused by changes in soil physical properties. The ecophysiological explanation of succession implies that: (1) fitness of a species for a particular succession stage at a particular location can be measured by stomatal behavior and biochemical assimilation capacity under local climate and soil conditions; (2) selection of species for a particular location at a particular succession stage can be guided by the parameter values provided in this study; and (3) succession may be accelerated by selecting trees with large root systems and large soil-root conductances that facilitate soil hydraulic redistribution of water. PMID- 12750058 TI - Effect of maternal chronic hypoxic exposure during gestation on apoptosis in fetal rat heart. AB - Chronic hypoxia during pregnancy is one of the most common insults to fetal development. We tested the hypothesis that maternal hypoxia induced apoptosis in the hearts of near-term fetal rats. Pregnant rats were divided into two groups, normoxic control and continuous hypoxic exposure (10.5% O2) from day 15 to 21 of gestation. Hearts were isolated from fetal rats of 21-day gestational age. Maternal hypoxia increased hypoxia-inducible factor-1alpha protein in fetal hearts. Chronic hypoxia significantly increased the percentage and size of binucleated myocytes and increased apoptotic cells from 1.4 +/- 0.14% to 2.7 +/- 0.3% in the fetal heart. In addition, the active cleaved form of caspase 3 was significantly increased in the hypoxic heart, which was associated with an increase in caspase 3 activity. There was a significant increase in Fas protein levels in the hypoxic heart. Chronic hypoxia did not change Bax protein levels but significantly decreased Bcl-2 proteins. In addition, chronic hypoxia significantly suppressed expression of heat shock protein 70. However, chronic hypoxia significantly increased expression of the anti-apoptotic protein 14-3-3, among other 14-3-3 isoforms. Chronic hypoxia differentially regulated beta adrenoreceptor (beta-AR) subtypes with an increase in beta1-AR levels but no changes in beta2-AR. The results demonstrate that maternal hypoxia increases apoptosis in fetal rat heart, which may be mediated by an increase in Fas and a decrease in Bcl-2 proteins. Chronic hypoxia-mediated increase in beta1-AR and decrease in heat shock proteins may also play an important role in apoptosis in the fetal heart. PMID- 12750059 TI - Metabolic and functional consequences of cytosolic 5'-nucleotidase-IA overexpression in neonatal rat cardiomyocytes. AB - Adenosine exerts a spectrum of energy-preserving actions on the heart negative chronotropic effects. The pathways leading to adenosine formation have remained controversial. In particular, although cytosolic 5'-nucleotidases can catalyze adenosine formation in cardiomyocytes, their contribution to the actions of adenosine has not been documented previously. We recently cloned two closely related AMP-preferring cytosolic 5'-nucleotidases (cN-IA and -IB); the A form predominates in the heart. In this study, we overexpressed pigeon cN-IA in neonatal rat cardiomyocytes using an adenovirus. cN-IA overexpression increased adenosine formation and release into the medium caused by simulated hypoxia and by isoproterenol in the absence and presence of inhibitors of adenosine metabolism. Adenosine release was not affected by an ecto-5'-nucleotidase inhibitor, alpha,beta-methylene-ADP, but was affected by a nucleoside transporter, dipyridamole. The positive chronotropic effect of isoproterenol (130 +/-3 vs. 100 +/-4 beats/min) was inhibited (107 +/-3 vs. 94 +/-3 beats/min) in cells overexpressing cN-IA, and this was reversed by the addition of the adenosine receptor antagonist 8-(p-sulfophenyl)theophilline (120 +/- 3 vs. 90 +/- 4 beats/min). Our results demonstrate that overexpressed cN-IA can be sufficiently active in cardiomyocytes to generate physiologically effective concentrations of adenosine at its receptors. PMID- 12750060 TI - Effect of nephrectomy and captopril on autoregulation of cerebral blood flow in rats. AB - The present study investigated the effect of circulating versus locally present renin on cerebral blood flow (CBF) and its autoregulation in rats. CBF was measured repetitively with the intracarotid 133Xe injection method, whereas blood pressure was lowered to determine the lower limit of autoregulation. To remove renin from the blood, rats were bilaterally nephrectomized and kept alive with peritoneal dialysis for 48 h. Five groups of animals were studied: 1) nephrectomized dialyzed rats, 2) nephrectomized dialyzed rats given a single intravenous dose of the angiotensin-converting enzyme inhibitor captopril (10 mg/kg), 3) sham nephrectomized and dialyzed rats, 4) rats receiving drugs as dialyzed rats but no surgery, and 5) rats given the same diet as the other groups but no drugs and no surgery. Baseline blood pressure was significantly lower in nephrectomized rats compared with controls. Nephrectomy, captopril, sham operation, or dialysis did not influence baseline CBF. The lower limit of CBF autoregulation was significantly lower in nephrectomized (53 +/- 4 mmHg) and sham operated (58 +/- 4 mmHg) rats compared with diet control rats (78 +/- 3 mmHg). Captopril significantly decreased the lower limit in nephrectomized rats (35 +/- 2 mmHg). Thus removal of circulating renin caused no change in the lower limit of autoregulation. By contrast, captopril lowered the lower limit even in the absence of circulating renin and hence appeared to exert its effect on components of the renin-angiotensin system in the cerebral resistance vessel walls. PMID- 12750061 TI - Subdiaphragmatic murine electrophysiological studies: sequential determination of ventricular refractoriness and arrhythmia induction. AB - Programmed electrical stimulation (PES) is a crucial aspect of the evaluation of the risk of arrhythmias in cardiac patients and provides a powerful tool for understanding the mechanisms of arrhythmia in experimental models. Whereas PES in the mouse is well characterized, the procedures allowing for follow-up studies in the same animal have not been developed. In this report, we describe a novel subdiaphragmatic approach that allows for repeat electrophysiological studies in the mouse. Under inhaled anesthesia, PES was performed in 36 wild-type mice via a stimulating electrode introduced through an epigastric incision and placed directly into the diaphragmatic surface of the heart. The procedure was repeated 7 days later. Ventricular effective refractory periods (VERP) did not change significantly between the initial and follow-up trials. Chronic treatment with amiodarone, however, was associated with a 70% prolongation in VERP from initial to follow-up studies (P < or = 0.001). In addition, PES of a genetically modified strain with sudden cardiac death, the connexin43 conditional knockout mouse consistently induced lethal polymorphic ventricular tachycardia. Thus sequential PES in mice is feasible with the use of a subdiaphragmatic approach, yields reproducible VERP values, and can be used to follow pharmacologically induced changes in VERP and identify mice at risk of lethal ventricular arrhythmias. PMID- 12750062 TI - Nitric oxide regulates retinal vascular tone in humans. AB - The purpose of the present study was to investigate the contribution of basal nitric oxide (NO) on retinal vascular tone in humans. In addition, we set out to elucidate the role of NO in flicker-induced retinal vasodilation in humans. Twelve healthy young subjects were studied in a three-way crossover design. Subjects received an intravenous infusion of either placebo or NG-monomethyl-L arginine (L-NMMA; 3 or 6 mg/kg over 5 min), an inhibitor of NO synthase. Thereafter, diffuse luminance flicker was consecutively performed for 16, 32, and 64 s at a frequency of 8 Hz. The effect of L-NMMA on retinal arterial and venous diameter was assessed under resting conditions and during the hyperemic flicker response. Retinal vessel diameter was measured with a Zeiss retinal vessel analyzer. L-NMMA significantly reduced arterial diameter (3 mg/kg: -2%; 6 mg/kg: 4%, P < 0.001) and venous diameter (3 mg/kg: -5%; 6 mg/kg: -8%, P < 0.001). After placebo infusion, flicker induced a significant increase in retinal vessel diameter (P < 0.001). At a flicker duration of 64 s, arterial diameter increased by 4% and venous diameter increased by 3%. L-NMMA did not abolish these hyperemic responses but blunted venous vasodilation (P = 0.017) and arterial vasodilation (P = 0.02) in response to flicker stimulation. Our data indicate that NO contributes to basal retinal vascular tone in humans. In addition, NO appears to play a role in flicker-induced vasodilation of the human retinal vasculature. PMID- 12750063 TI - Renal vascular responses to static handgrip: role of muscle mechanoreflex. AB - During exercise, the sympathetic nervous system is activated, which causes vasoconstriction. The autonomic mechanisms responsible for this vasoconstriction vary based on the particular tissue being studied. Attempts to examine reflex control of the human renal circulation have been difficult because of technical limitations. In this report, the Doppler technique was used to examine renal flow velocity during four muscle contraction paradigms in conscious humans. Flow velocity was divided by mean arterial blood pressure to yield an index of renal vascular resistance (RVR). Fatiguing static handgrip (40% of maximal voluntary contraction) increased RVR by 76%. During posthandgrip circulatory arrest, RVR remained above baseline (2.1 +/- 0.2 vs. 2.8 +/- 0.2 arbitrary units; P < 0.017) but was only 40% of the end-grip RVR value. Voluntary biceps contraction increased RVR within 10 s of initiation of contraction. This effect was not associated with an increase in blood pressure. Finally, involuntary biceps contraction also raised RVR. We conclude that muscle contraction evokes renal vasoconstriction in conscious humans. The characteristic of this response is consistent with a primary role for mechanically sensitive afferents. This statement is based on the small posthandgrip circulatory arrest response and the vasoconstriction that was observed with involuntary biceps contraction. PMID- 12750064 TI - Attenuation of activity-induced increases in cerebellar blood flow by lesion of the inferior olive. AB - We sought to define the contribution of the climbing fibers (CF), one of the major inputs to Purkinje neurons, to the increase in cerebellar blood flow (BFcrb) produced by activation of the cerebellar cortex. The neurotoxin 3 acetylpyridine was used to lesion the inferior olive, the site from which the CF originate. Crus II, an area of the cerebellar cortex that receives sensory afferents from the perioral region, was activated by low-intensity stimulation of the upper lip (5-25 V and 4-16 Hz) in sham-lesioned and lesioned mice. BFcrb was recorded in crus II using a laser-Doppler flow probe. The increase in BFcrb produced by harmaline, an alkaloid that activates the CF, was abolished in lesioned mice (P > 0.05 vs. BFcrb before harmaline, n = 6), attesting to the effectiveness of the lesion. In sham-lesioned animals, upper lip stimulation increased BFcrb in crus II by 25 +/- 2% (25 V and 10 Hz, n = 6). The rise in BFcrb was attenuated by 63 +/- 7% (25 V and 10 Hz) in lesioned mice (P < 0.05, n = 6). In contrast, the increase in BFcrb produced by hypercapnia was not affected (P > 0.05). These data suggest that CF are responsible for a substantial portion of the increase in BFcrb produced by crus II activation. Thus the hemodynamic response evoked by functional activation of the cerebellar cortex reflects, in large part, CF activity. PMID- 12750065 TI - Intracellular proton mobility and buffering power in cardiac ventricular myocytes from rat, rabbit, and guinea pig. AB - Intracellular pH (pHi) is an important modulator of cardiac function. The spatial regulation of pH within the cytoplasm depends, in part, on intracellular H+ (Hi+) mobility. The apparent diffusion coefficient for Hi+, DHapp, was estimated in single ventricular myocytes isolated from the rat, guinea pig, and rabbit. DHapp was derived by best-fitting predictions of a two-dimensional model of H+ diffusion to the local rise of intracellular [H+], recorded confocally (ratiometric seminaphthorhodafluor fluorescence) downstream from an acid-filled, whole cell patch pipette. Under CO2/HCO3--free conditions, DHapp was similar in all three species (mean values: 8-12.5 x 10-7 cm2/s) and was over 200-fold lower than that for H+ in water. In guinea pig myocytes, DHapp was increased 2.5-fold in the presence of CO2/HCO3- buffer, in agreement with previous observations in rabbit myocytes. Hi+ mobility is therefore low in cardiac cells, a feature that may predispose them to the generation of pHi gradients in response to sarcolemmal acid/base transport or local cytoplasmic acid production. Low Hi+ mobility most likely results from H+ shuttling among cytoplasmic mobile and fixed buffers. This hypothesis was explored by comparing the pHi dependence of intrinsic, intracellular buffering capacity, measured for all three species, and subdividing buffering into mobile and fixed fractions. The proportion of buffer that is mobile will be the main determinant of DHapp. At a given pHi, this proportion appeared to be similar in all three species, consistent with a common value for DHapp. Over the pHi range of 6.0-8.0, the proportion is expected to change, predicting that DHapp may display some pHi sensitivity. PMID- 12750066 TI - Inhaled NO inhibits platelet aggregation and elevates plasma but not intraplatelet cGMP in healthy human volunteers. AB - Effects of inhaled nitric oxide (NO) on human platelet function are controversial. It is uncertain whether intraplatelet cGMP mediates the effect of inhaled NO on platelet function. We investigated the effect of 30 ppm inhaled NO on platelet aggregation and plasma and intraplatelet cGMP in 12 subjects. We performed platelet aggregation studies by using a photooptical aggregometer and five agonists (ADP, collagen, epinephrine, arachidonic acid, and ristocetin). During inhalation, the maximal extent of platelet aggregation decreased by 75% with epinephrine (P < 0.005), 56% with collagen (P < 0.005), and 20% with arachidonic acid (P < 0.05). Responses to ADP (8% P > 0.05) and ristocetin (5% P > 0.05) were unaffected. Platelet aggregation velocity decreased by 64% with collagen (P < 0.005), 60% with epinephrine (P < 0.05), 33% with arachidonic acid (P < 0.05), and 14% with ADP (P > 0.05). Plasma cGMP levels increased from 2.58 +/- 0.43 to 9.99 +/- 5.57 pmol/ml (P < 0.005), intraplatelet cGMP levels were unchanged (means +/- SD: 1.96 +/- 0.58 vs. 2.71 +/- 1.67 pmol/109 platelets; P > 0.05). Inhaled NO inhibits platelet aggregation via a cGMP independent mechanism. PMID- 12750067 TI - Role of microtubules versus myosin heavy chain isoforms in contractile dysfunction of hypertrophied murine cardiocytes. AB - In large mammals there is a correlation between microtubule network densification and contractile dysfunction in severe pressure-overload hypertrophy. In small mammals there is a similar correlation for the shift to beta-myosin heavy chain (MHC), a MHC isoform having a slower ATPase Vmax. In this study, murine left ventricular (LV) pressure overload invoked both mechanisms: microtubule network densification and beta-MHC expression. Cardiac beta-MHC was also augmented without altering tubulin levels by two load-independent means, chemical thyroidectomy and transgenesis. In hypertrophy, contractile function of the LV and its cardiocytes decreased proportionally; microtubule depolymerization restored normal cellular contraction. In hypothyroid mice having a complete shift from alpha-MHC to beta-MHC, contractile function of the LV and its cardiocytes also decreased, but microtubule depolymerization had no effect on cellular contraction. In transgenic mice having a cardiac beta-MHC increase similar to that in hypertrophy, contractile function of the LV and its cardiocytes was normal, and microtubule depolymerization had no effect. Thus, although both mechanisms may cause contractile dysfunction, for the extent of MHC isoform switching seen even in severe murine LV pressure-overload hypertrophy, microtubule network densification appears to have the more important role. PMID- 12750068 TI - Attenuation of chronic hypoxic pulmonary hypertension by simvastatin. AB - The 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) have been shown to improve multiple normal endothelial cell functions and inhibit vascular wall cell proliferation. We hypothesized that one such agent, simvastatin, would attenuate chronic hypoxic pulmonary hypertension. Male adult Sprague-Dawley rats were exposed (14 days) to normoxia (N), normoxia plus once-a day administered simvastatin (20 mg/kg ip) (NS), hypoxia (10% inspired O2 fraction) (H), or hypoxia plus simvastatin (HS). Mean pulmonary artery pressure, measured in anesthetized, ventilated rats with an open-chest method, was reduced from 25 +/- 2 mmHg in H to 18 +/- 1 in HS (P < 0.001) but did not reach normoxic values (12 +/- 1 mmHg). Similarly, right ventricular/left ventricular plus interventricular septal weight was reduced from 0.53 +/- 0.02 in the H group to 0.36 +/- 0.02 in the HS group (P < 0.001). The increased hematocrit in H (0.65 +/ 0.02) was prevented by simvastatin treatment (0.51 +/- 0.01, P < 0.001). Hematocrit was similar in N versus NS. Alveolar vessel muscularization and medial thickening of vessels 50-200 microM in diameter induced by hypoxia were also significantly attenuated in the HS animals. Lung endothelial nitric oxide synthase (eNOS) protein expression in the HS group was less than H (P < 0.01) but was similar in N versus NS. We conclude that simvastatin treatment potently attenuates chronic hypoxic pulmonary hypertension and polycythemia in rats and inhibits vascular remodeling. Enhancement of lung eNOS expression does not appear to be involved in mediating this effect. PMID- 12750070 TI - Regarding the surgical reversal of presbyopia (SRP) surgery for presbyopia, vol 108, number 12, December 2001, 2161-2. PMID- 12750069 TI - Dobutamine potentiates arterial chemoreflex sensitivity in healthy normal humans. AB - beta-Adrenergic agonists may increase chemosensitivity in humans. We tested the hypothesis that the beta1-agonist dobutamine increases peripheral chemosensitivity in a double-blind placebo-controlled randomized and crossover study. In 15 healthy subjects, we examined the effects of dobutamine on breathing, hemodynamics, and sympathetic nerve activity (measured using microneurography) during normoxia, isocapnic hypoxia (10% O2), posthypoxic maximal voluntary end-expiratory apnea, hyperoxic hypercapnia, and cold pressor test (CPT). Dobutamine increased ventilation (7.5 +/- 0.3 vs. 6.7 +/- 0.2 l/min, P = 0.0004) during normoxia, markedly enhanced the ventilatory (16.1 +/- 1.6 vs. 11.4 +/- 0.7 l/min, P < 0.0001) and sympathetic (+403 +/- 94 vs. +222 +/- 5%, P < 0.03) responses at the fifth minute of isocapnic hypoxia, and enhanced the sympathetic response to the apnea performed after hypoxia (+501 +/- 107% vs. +291 +/- 38%, P < 0.05). No differences were observed between dobutamine and placebo on the responses to hyperoxic hypercapnia and CPT. Dobutamine increases ventilation during normoxia and potentiates the ventilatory and sympathetic responses to hypoxia in healthy subjects. Dobutamine does not affect the responses to hyperoxic hypercapnia and CPT. We conclude that dobutamine enhances peripheral chemosensitivity. PMID- 12750072 TI - Regarding the surgical reversal of presbyopia (SRP) surgery for presbyopia, vol 108, number 12, December 2001, 2161-2. PMID- 12750073 TI - Causes of diffuse lamellar keratitis. PMID- 12750075 TI - Technology assessment of cyclophotocoagulation. PMID- 12750077 TI - Polar strain and crystalline lens age. PMID- 12750079 TI - Primary ophthalmic rhabdomyosarcoma. PMID- 12750081 TI - Excessive blinking in children. PMID- 12750083 TI - Anti-vascular endothelial growth factor therapy for neovascular age-related macular degeneration: promises and pitfalls. PMID- 12750084 TI - Viscocanalostomy versus trabeculectomy in white adults affected by open-angle glaucoma: a 2-year randomized, controlled trial. AB - PURPOSE: To compare the effectiveness and safety of viscocanalostomy and trabeculectomy in adults with uncontrolled open-angle glaucoma. DESIGN: Single masked, parallel-group, prospective, randomized 24-month trial, with 90% power to detect a clinically important difference between groups. PARTICIPANTS: Fifty consecutive patients (50 eyes) with primary open-angle or pseudoexfoliative glaucoma. INTERVENTION: Eyes were assigned randomly to either viscocanalostomy (group 1) or trabeculectomy (group 2) with no intraoperative antifibrotics in the study eye. In group 1, no further intervention was allowed, whereas trabeculectomy eyes could receive subconjunctival 5-fluorouracil (5-FU) injections or laser suture lysis after surgery. MAIN OUTCOME MEASURES: Success rate based on intraocular pressure (IOP), visual acuity, discomfort, and other complications. RESULTS: At the end of the 24-month follow-up, IOP of 21 mmHg or less and more than 6 mmHg was achieved in 76% in group 1 (n = 19) and in 80% in group 2 (n = 20; log-rank P = 0.60); an IOP between 6 and 16 mmHg was obtained in 56% in group 1 (n = 14) and in 72% in group 2 (n = 18; log-rank P = 0.17; Kaplan Meier cumulative probability of success). Complications of viscocanalostomy included one intraoperative conversion into trabeculectomy; microruptures in Descemet's membrane in five eyes; three cases of iris incarceration in the Decemet's window, two of which caused early failure of the procedure requiring reoperation; and a 1-mm to 2-mm transient self-resolving hyphema in three cases. Complications of trabeculectomy included one case of postoperative bleb bleeding with early transient IOP spike; one early hyphema; five cases of postoperative hypotony, two of which had a positive Seidel test from the conjunctival suture; three cases of transient choroidal detachment, two of which had shallow anterior chamber. No patient required reoperation. Two eyes required argon laser suture lysis, and nine underwent one or more 5-FU injections, which caused punctate keratopathy in three eyes. CONCLUSIONS: Viscocanalostomy is an effective IOP lowering procedure in white adults affected by open-angle glaucoma. Trabeculectomy with postoperative 5-FU can probably provides lower IOPs but, with more numerous complications, greater discomfort, and more intensive postoperative management. PMID- 12750085 TI - Intermediate-term outcome of Baerveldt glaucoma implants in Asian eyes. AB - PURPOSE: To determine the intermediate-term efficacy and safety of Baerveldt glaucoma implants in Asian eyes with complicated glaucoma. DESIGN: Retrospective, nonrandomized, comparative trial. PARTICIPANTS: One hundred twenty-four Asian patients (124 eyes) with complicated glaucoma. INTERVENTION: Implantation of 54 250-mm(2) and 70 350-mm(2) Baerveldt glaucoma drainage implants at Singapore National Eye Center from 1994 through 1999. MAIN OUTCOME MEASURES: Intraocular pressure, number of glaucoma medications, and complications. RESULTS: The mean follow-up period was 33.4 +/- 14.4 months (mean +/- SD; range, 12-72 months). Intraocular pressure (IOP) was reduced from a mean preoperative IOP of 36.5 +/- 10.7 mmHg (range, 21-80 mmHg) to 15.3 +/- 6.0 mmHg (range, 1-34 mmHg), and the number of glaucoma medications decreased from 2.6 +/- 0.6 (range, 1-5) before the time of surgery to 0.5 +/- 0.9 (range, 0-4) medications at last follow-up. Overall, there were 67 eyes (54%) that were classified as complete successes, 27 eyes (22%) that were qualified successes, and 30 eyes that failed (24%). Postoperative complications occurred in 43 eyes (34.7%), and 21 eyes (17%) required further surgical intervention and revision. There was no statistically significant difference between the 250-mm(2) and 350-mm(2) type of implants in terms of success rates, final IOP, number of medications, and rates of complications. After adjusting for gender, preoperative IOP, and length of follow up, increasing age (odds ratio [OR], 1.05; 95% confidence interval [CI], 1.01 1.09; P = 0.02) and the number of previous operations performed before implant surgery (OR, 1.57; 95% CI, 1.07-2.31; P = 0.01) were found to be correlated positively with failure. CONCLUSIONS: In Asian eyes with complicated glaucoma, Baerveldt glaucoma implants achieve stable and satisfactory IOP reduction with low incidence of complications in the intermediate term after surgery. PMID- 12750086 TI - Long-term results of noncontact neodymium:yttrium-aluminum-garnet cyclophotocoagulation in neovascular glaucoma. AB - PURPOSE: To determine the long-term efficacy and safety of noncontact transscleral neodymium:yttrium-aluminum-garnet (Nd:YAG) cyclophotocoagulation (CP) for the treatment of neovascular glaucoma (NVG). DESIGN: Retrospective, noncomparative, interventional case series. PARTICIPANTS: One hundred fifteen eyes of 111 subjects treated from December 1987 to January 2001. METHODS: Eyes with uncontrolled NVG underwent noncontact Nd:YAG CP. Treatment parameters and pretreatment and posttreatment intraocular pressures (IOP) were reviewed. Preoperative and postoperative IOP were compared using a paired Student's t test. Success was defined as an IOP /=40 years. PARTICIPANTS: A total of 2996 persons without open-angle glaucoma or receiving IOP-lowering medication at baseline. METHODS: Participants underwent standardized examinations including applanation tonometry, measurement of blood pressure, and anthropometric indices; a detailed interview; various ocular measurements; and venipuncture for glycosylated hemoglobin (GHb). Diabetes was defined by self reported physician diagnosis and hypertension by blood pressure >/=140/90 mmHg and/or treatment history. MAIN OUTCOME MEASURES: The 4-year person-based IOP change between baseline and follow-up was defined as the more positive IOP difference in either eye. RESULTS: An IOP >21 mmHg at baseline was more likely in black and in mixed (black and white) participants (age-gender adjusted odds ratio [OR], 3.9 and 3.8, respectively) than in whites. Similarly, these groups had more hypertension (age-gender adjusted OR, 2.4 and 2.1, respectively) and diabetes (age-gender adjusted OR, 3.9 and 1.7, respectively) than did whites. Mean IOP in black participants increased by 2.5 (standard deviation, 3.9) mmHg over 4 years. Multiple regression analyses showed that baseline diabetes history and hypertension, as well as older age, elevated GHb, higher blood pressures, and lower baseline IOP were associated with a 4-year increase of IOP. The association between diabetes history/GHb and IOP increase became borderline/nonsignificant when persons who underwent cataract surgery during follow-up were excluded. CONCLUSIONS: This report provides new data on the relationship of systemic factors to longitudinal increases in IOP in an African-origin population. Results highlight the increased risk of elevated IOP in populations with high prevalences of diabetes and hypertension. PMID- 12750089 TI - Correlation between intraocular pressure level and optic disc changes in high tension glaucoma suspects. AB - PURPOSE: To assess the correlation between intraocular pressure (IOP) and future optic disc changes in eyes of patients suspected of having high-tension glaucoma and to determine the target pressure in eyes of patients suspected of having high tension glaucoma. DESIGN: Prospective, comparative, observational case series. PARTICIPANTS: Fifty-seven eyes of 57 patients suspected of having high-tension glaucoma selected from 226 consecutive patients with glaucoma or suspected glaucoma. INTERVENTION: The status of the optic disc and visual fields was evaluated during an initial examination by confocal laser ophthalmoscopy and automatic perimetry, respectively. Fifty-seven eyes were followed up for at least 2.5 years and then underwent a final examination. IOP was measured every 3 months. MAIN OUTCOME MEASURES: The correlation between changes in optic disc status and IOP control during follow-up was determined. Changes in the optic disc were evaluated by using the percentage change in total contour area, neuroretinal rim area, and cup-to-disc ratio. The control IOPs were evaluated on the basis of the percentage of time during which IOP was <18 mmHg (%<18 mmHg control) and <21 mmHg (%<21 mmHg control) during follow-up in each eye. The correlation between visual field changes and IOP control was also analyzed. RESULTS: Results from 48 (84.2%) of 57 participants were included. Patients were followed up for a mean 4.4 years. The %<21 mmHg control was significantly correlated with changes in neuroretinal rim area (r = 0.40; P = 0.0055) and in cup-to-disc ratio (r = -0.40; P = 0.0051) by linear regression analysis. The correlations were also significant if the difference in the length of follow-up was adjusted by multivariate regression analysis. There was no significant correlation between %<18 mmHg control and changes in disc parameters. Seven (14.6%) of 48 eyes had glaucomatous visual field defects at the final examination. The %<21 mmHg control was significantly lower in eyes with visual field changes compared with eyes with no changes (P = 0.0153; unpaired t test). In contrast, %<18 mmHg control was not significantly different between eyes with and without visual field changes (P = 0.3886). CONCLUSIONS: The IOP level correlates with topographic changes in the optic disc in eyes of patients suspected of having high-tension glaucoma. The target pressure for such eyes may need to be between 18 and 21 mmHg, and <18 mmHg is a safe target level in the treatment of patients suspected of having high tension glaucoma to delay topographic optic disc changes. PMID- 12750090 TI - Helicobacter pylori infection and the risk for open-angle glaucoma. AB - PURPOSE: To determine the frequency of exposure to Helicobacter pylori infection in glaucoma patients. DESIGN: Prospective case-control study. PARTICIPANTS: Ninety-seven consecutive patients attending a glaucoma clinic. These included 38 patients with primary open-angle glaucoma (POAG), 19 with normal pressure glaucoma (NPG), 16 with pseudoexfoliation glaucoma (PXE), and 24 with ocular hypertension (OHT). Ninety-four age-matched participants without glaucoma served as a control population. METHODS: Serum was analyzed for the presence of H. pylori-specific immunoglobulin G antibodies by enzyme-linked immunosorbent assay. MAIN OUTCOME MEASURES: Serologic analysis for H. pylori. RESULTS: Seropositivity for H. pylori was higher in patients with glaucoma (26.0%) than in controls (20.2%), but this did not achieve statistical significance (P = 0.46). A total of 26.3% of POAG patients, 26.3% of NPG patients, 25.0% of PXE patients, and 25.0% of OHT patients were seropositive. CONCLUSIONS: This study suggests that exposure to H. pylori infection is not associated with open-angle glaucoma. PMID- 12750091 TI - Antibiotics for toxoplasmic retinochoroiditis: an evidence-based systematic review. AB - PURPOSE: To determine the effectiveness of systemic antibiotic treatment for toxoplasmic retinochoroiditis. CLINICAL RELEVANCE: Toxoplasma retinochoroiditis is a significant cause of visual morbidity. Multiple different antibiotic regimens are used, but controversy about treatment effectiveness remains. LITERATURE REVIEWED: Searches were conducted of Cochrane Controlled Trials Register, Medline (1966 onward), Embase (1980 onward), Dissertation Abstracts (1861 onward), Lilacs (1982 onward), and Pascal (1984 onward). Pharmaceutical companies were contacted for unpublished data. Any randomized controlled trials that compared antibiotics versus placebo in immunocompetent patients with toxoplasmic retinochoroiditis were retrieved. Primary outcome measures were long term visual acuity and risk of recurrent retinochoroiditis. Secondary outcomes included duration and severity of acute symptoms, size of the lesion at end of follow-up, and adverse effects of treatment. RESULTS: Only 3 studies (total of 173 participants) were randomized controlled trials and hence met the inclusion criteria (level II). All 3 were methodologically poor, and 2 were carried out more than 35 years ago. None reported the effect on long-term visual outcome. We found no evidence for a beneficial effect on the duration and severity of signs of acute toxoplasmic retinochoroiditis (A,II). There was weak evidence for an effect of long-term treatment for chronic recurrent toxoplasmic retinochoroiditis on lesion recurrence. Treatment was associated with adverse effects. CONCLUSIONS: There is a lack of evidence to support routine antibiotic treatment for acute toxoplasmic retinochoroiditis. Placebo-controlled randomized trials of antibiotic treatment in patients presenting with acute or chronic toxoplasmic retinochoroiditis arising in any part of the retina are required. PMID- 12750093 TI - Retinal microvascular abnormalities and 10-year cardiovascular mortality: a population-based case-control study. AB - PURPOSE: Retinal microvascular abnormalities reflect persistent arteriolar damage from hypertension and independently predict stroke. We examined their associations with long-term cardiovascular mortality. DESIGN: Population-based, nested, case-control study. POPULATION: Cases were Beaver Dam Eye Study participants (age range, 43-84 years) who died of coronary heart disease or stroke between the baseline examination in 1988 to 1990 and 1999 (n = 413). Nearly 3 controls per case were selected from the baseline cohort, frequency matched on 5-year age intervals and gender (n = 1198). METHODS: Retinal photographs of cases and controls at baseline were evaluated for retinopathy, focal arteriolar narrowing, and arteriovenous nicking by graders masked to case control status using standardized protocols. To obtain an estimate of generalized arteriolar narrowing, photographs were digitized and diameters of individual retinal vessels were measured and summarized by a computer program. MAIN OUTCOME MEASURE: Ten-year cardiovascular mortality. RESULTS: After controlling for systolic blood pressure, diabetes, glycosylated hemoglobin levels, and other risk factors, retinopathy was associated with increased cardiovascular mortality, with odds ratios of 1.8 (95% confidence interval [CI], 1.2, 2.7). For other retinal abnormalities, associations with cardiovascular mortality were present only in younger people, with odds ratios of 2.7 (95% CI, 1.0, 7.4) for focal arteriolar narrowing, 1.8 (95% CI, 0.8, 4.5) for arteriovenous nicking, and 1.9 (95% CI, 1.2, 2.9) for generalized arteriolar narrowing in persons 43 to 74 years of age but odds ratios of 1.1, 0.4, and 1.0 for the corresponding retinal abnormalities in persons 75 years and older. CONCLUSIONS: Retinopathy is independently associated with cardiovascular mortality. Associations for other retinal abnormalities were only observed in middle-aged persons. These data support recent studies that suggest retinal microvascular abnormalities provide independent information regarding cardiovascular risk. PMID- 12750094 TI - Incidence of diabetic retinopathy in the Barbados Eye Studies. AB - PURPOSE: To examine the 4-year incidence and risk factors for diabetic retinopathy (DR) among black participants with diabetes in the Barbados Eye Studies (BES). DESIGN: Population-based incidence study. SETTING AND PARTICIPANTS: Four hundred ten persons with diabetes mellitus (DM) from the BES cohort, which was based on a simple random sample of Barbadians, 40 to 84 years of age at baseline. MAIN OUTCOME MEASURES: Development of DR, assessed by independent gradings of 30 degrees color stereo fundus photographs of the disc and macula. Associations were evaluated by logistic regression analyses. RESULTS: After 4 years, DR developed in 92 of 306 (30.1%; 95% confidence interval, 25.0%, 35.5%) persons unaffected at baseline. The incidence of DR was 31.9% in those with known DM at baseline and 20.9% in newly diagnosed DM. Clinically significant macular edema developed in 16 (4.5%) of 353 individuals at risk. Seven (6.9%) of the 101 persons with minimum or moderate DR at baseline progressed to proliferative DR. Age-specific incidence declined from 36.2% at age 40 to 49 years to 28.8% and 24.2% over the subsequent two decades, increasing to 38.2% among those >/=70 years. Risk factors for DR were increased systolic blood pressure (relative risk [RR], 1.16 [1.03, 1.31]/10 mmHg increase); use of oral hypoglycemics (RR, 2.4 [1.3, 4.2]); and use of insulin (RR, 6.1 [1.7, 22.1]) (vs. no treatment or diet only); and elevated glycated hemoglobin (GHb; RR, 6.4 [2.5, 16.0]); GHb >11.5% vs. GHb /=10 large (>63- micro m) drusen in the contralateral, or fellow, eye. INTERVENTION: The fellow eye of 59 patients was assigned randomly to argon green laser treatment consisting of multiple 100- micro m spots at least 750 micro m from the center of the fovea. The fellow eye of the remaining 61 patients was assigned randomly to observation. MAIN OUTCOME MEASURES: Change in visual acuity was the primary outcome measure. Incidence of choroidal neovascularization, resolution of drusen, change in contrast threshold, change in critical print size for reading, and incidence of geographic atrophy were secondary outcome measures. RESULTS: Throughout 4 years of follow-up, there were no statistically significant differences in change in visual acuity, contrast threshold, critical print size, or incidence of geographic atrophy. With additional follow-up, the large increase in the incidence of choroidal neovascularization observed within 18 months of treatment was maintained; however, by 30 months, the incidence in the two treatment groups was the same. Most drusen resolution in treated eyes occurred within 24 months of the initial treatment. Treated eyes that received higher-intensity laser burns had an increased risk of choroidal neovascularization. Among eyes developing choroidal neovascularization in each treatment group, most lesions (two thirds or more) were composed of occult neovascularization only. CONCLUSIONS: Laser treatment as applied in the CNVPT caused an excess risk of choroidal neovascularization in the first year or so after treatment. The increased early incidence of choroidal neovascularization was not associated with either a harmful or beneficial effect in this pilot study. PMID- 12750101 TI - Anti-vascular endothelial growth factor therapy for subfoveal choroidal neovascularization secondary to age-related macular degeneration: phase II study results. AB - PURPOSE: There is evidence to suggest that anti-vascular endothelial growth factor (anti-VEGF) therapy may be useful in treating ocular neovascularization. A phase IA single intravitreal injection study of anti-VEGF therapy for patients with subfoveal choroidal neovascularization (CNV) secondary to age-related macular degeneration (AMD) revealed a good safety profile. We performed a phase II multiple injection study of anti-VEGF therapy with and without photodynamic therapy for patients with subfoveal CNV secondary to AMD to determine the safety profile of multiple injection therapy. DESIGN: A phase II multiple-dose safety study. PARTICIPANTS/METHODS: Twenty-one patients were treated with intravitreal injection with and without photodynamic therapy. MAIN OUTCOME MEASURES: Clinical evidence of toxicity and complications. RESULTS: No drug-related serious adverse events were revealed. Ophthalmic evaluation revealed that 87.5% of patients who received the anti-VEGF aptamer alone showed stabilized or improved vision 3 months after treatment and that 25% of eyes demonstrated a 3 line or greater improvement in vision on the Early Treatment of Diabetic Retinopathy Study chart during this period. A 60% 3 line gain at 3 months was noted in patients who received both the anti-VEGF aptamer and photodynamic therapy. CONCLUSIONS: Anti VEGF therapy is a promising treatment for various forms of ocular neovascularization, including AMD. Multiple intravitreal injections of the anti VEGF aptamer were well tolerated in this phase II study. Further clinical trials are necessary to demonstrate the efficacy and long-term safety of anti-VEGF therapy for AMD. PMID- 12750102 TI - The effect of cytomegalovirus retinitis on the quality of life of patients with AIDS in the era of highly active antiretroviral therapy. AB - PURPOSE: To evaluate the effect of cytomegalovirus (CMV) retinitis on quality of life (QOL) in patients with the acquired immune deficiency syndrome (AIDS). DESIGN: Enrollment data from a multicenter, prospective cohort study. PARTICIPANTS: Patients >/=13 years of age with AIDS in three groups: no CMV retinitis, 709 patients; long-standing CMV retinitis (diagnosed a median of 34.5 months previously), 212 patients; and newly diagnosed CMV retinitis (diagnosed /=5/200 was achieved in 67.5%. CONCLUSIONS: Current vitreoretinal surgical techniques enable removal of intraocular cysticercosis in all cases, with reattachment of the retina in 86.6% and recovery of ambulatory vision in approximately 67% of cases. PMID- 12750104 TI - Congenital simple hamartoma of the retinal pigment epithelium: a study of five cases. AB - PURPOSE: To report 5 patients with presumed congenital, simple hamartoma of the retinal pigment epithelium (RPE). DESIGN: Retrospective, observational small case series. PARTICIPANTS: Five patients with a black macular tumor of the RPE. MAIN OUTCOME MEASURES: Initial tumor features, initial associated fundus features, tumor course on follow-up, and visual acuity outcome. RESULTS: In all 5 cases, the tumor involved the macula and the mean distance to the foveola was 0.4 mm (median, 0.2 mm; range, 0.2-1.2 mm). In all cases, the tumor was darkly pigmented, with full-thickness retinal and RPE involvement and minimal protrusion into the vitreous cavity. The mean ultrasonographic tumor thickness was 1.6 mm (median, 1.4 mm; range, 1.1-2.5 mm). Associated features included minimally dilated retinal feeding artery and draining vein (100%), surrounding mild retinal traction (80%), retinal exudation (20%), and vitreous pigmented cells (20%). None of the lesions had associated subretinal fluid, macular edema, or macular hole. Visual acuity was 20/20 in 3 cases and slightly decreased in the other 2 cases because of foveal traction. The findings remained stable in 3 patients who had adequate follow-up. We chose to classify this lesion as congenital simple hamartoma of the RPE to differentiate it from combined hamartoma of the retina and RPE, congenital hypertrophy of the RPE, adenoma or adenocarcinoma of the RPE, and acquired hyperplasia of the RPE, which are different clinical entities. CONCLUSIONS: Congenital hamartoma of the RPE is typically a black, full-thickness retinal mass, often adjacent to the foveola. Despite such location, the visual acuity was affected minimally. PMID- 12750105 TI - Subconjunctival mitomycin C as adjunctive therapy before pterygium excision. AB - PURPOSE: To evaluate the safety and efficacy of subconjunctival mitomycin C as adjunctive therapy before pterygium surgery. DESIGN: Prospective noncomparative case series. PARTICIPANTS: Thirty-six eyes of 36 patients. INTERVENTION: Thirty six eyes of 36 patients prospectively received 0.1 ml of 0.15 mg/ml mitomycin C subconjunctivally injected into the head of the pterygium 1 month before bare sclera surgical excision. MAIN OUTCOME MEASURE: Recurrence of pterygia. RESULTS: The pterygia resolved in 34 (94%) of 36 eyes, with a recurrence rate of 6% over a mean follow-up of 24.4 months. No wound-healing complication developed in any patient. CONCLUSIONS: Subconjunctival mitomycin C is an effective treatment before pterygium excision. Subconjunctival injection allows exact titration of mitomycin C delivery to the activated fibroblasts and minimizes epithelial toxicity. PMID- 12750106 TI - Long-term outcomes of combined penetrating keratoplasty with iris-sutured intraocular lens implantation. AB - PURPOSE: To evaluate the long-term outcomes of combined penetrating keratoplasty with iris-sutured posterior chamber intraocular lens implantation. PARTICIPANTS: Patients (264 eyes) with aphakic and pseudophakic bullous keratopathy. DESIGN: Retrospective case series. INTERVENTION: Penetrating keratoplasty combined with implantation of an iris-sutured posterior chamber intraocular lens performed by a single surgeon. METHODS: Data were gathered from the institutional database during a period of 10 years (1989-1999) and analyzed using customized software. MAIN OUTCOME MEASURES: Graft survival rate, visual acuity, and intraoperative and postoperative complications. RESULTS: Two-hundred fifty-two consecutive patients (264 eyes) were operated. Graft survival rates were 95% at 1 year, 89% at 2 years, and 81% at 5 years and were comparable to the results of previous corneal transplant studies. Among 180 patients (191 eyes) with a follow-up of at least 2 years (range, 2-11 years; mean, 5 years), the visual acuity at final follow-up visit, compared with preoperative visual acuity was improved in 72% of eyes (mean preoperative visual acuity, 20/250; mean postoperative visual acuity, 20/60). No intraoperative complications were encountered. CONCLUSIONS: Combined penetrating keratoplasty with iris-sutured posterior chamber intraocular lens implantation offers significant vision benefits in this patient group. PMID- 12750107 TI - The effect of hinge position on corneal sensation and dry eye after LASIK. AB - PURPOSE: To investigate the effect of hinge position on corneal sensation and dry eye syndrome after laser in situ keratomileusis (LASIK). DESIGN: Prospective, randomized, self-controlled trial. PARTICIPANTS: Fifty-two patients >/=18 years of age undergoing bilateral LASIK. INTERVENTION: Patients underwent bilateral LASIK with the superior-hinge Hansatome microkeratome in one eye and the nasal hinge Amadeus microkeratome in the other eye. In all eyes, the flaps were 160 micro m thick, with a diameter of 9.5 mm. MAIN OUTCOME MEASURES: Masked Cochet Bonnet esthesiometry was performed centrally before surgery and at 1 week, 1 month, 3 months, and 6 months after surgery. Dry eye was evaluated at the same time intervals with lissamine green corneal and conjunctival staining, Schirmer testing with anesthesia, and tear-film breakup time. Subjective evaluation of dry eye sensation was performed at 3 and 6 months after surgery. RESULTS: Corneal sensation was reduced in eyes with either superior- or nasal-hinge corneal flaps at 1 week, 1 month, and 3 months after surgery (P < 0.001). Compared with preoperative values, a significant reduction in corneal sensation remained at 6 months in corneas with superior-hinge flaps (P < 0.001) but not in corneas with nasal-hinge flaps (P = 0.263). Mean corneal sensation was greater in corneas with a nasal-hinge flap compared with corneas with a superior-hinge flap at all postoperative visits (P < 0.001). The loss of sensation was greatest at 1 week and showed improvement at each subsequent time interval up to 6 months. Overall, dry eye signs and symptoms were greatest during the immediate postoperative period and improved at all subsequent time intervals. Dry eye signs and symptoms were generally greatest in the eyes with a superior-hinge flap and milder in eyes with a nasal-hinge flap. CONCLUSIONS: The long posterior corneal nerves, which innervate the cornea, enter the eye at 3- and 9-o'clock. A superior-hinge flap transects both arms of the neuroplexus, whereas a nasal hinge transects only the temporal arm. LASIK results in a significant reduction in corneal sensation. Corneal sensation and dry eye signs and symptoms decreased immediately after LASIK and improved at all time periods between 1 week and 6 months in eyes with both a nasal-hinge flap and a superior-hinge flap. However, the loss of corneal sensation and presence of dry eye syndrome were greater in eyes with a superior hinge flap than in eyes with a nasal-hinge flap. PMID- 12750109 TI - Intacs for keratoconus. AB - PURPOSE: To evaluate the efficacy and safety of placement of Intacs in subjects with keratoconus. DESIGN: Retrospective, nonrandomized comparative trial. INTERVENTION: Intrastromal corneal ring segment implantation. PARTICIPANTS: Seventy-four eyes of 50 subjects (41 male and 9 female) were evaluated. The mean age of subjects in the study was 35 years, ranging from 20 to 73 years. Twenty six subjects underwent single-eye treatment, and 24 subjects had both eyes treated. METHODS: A modified Intacs procedure was performed on subjects with keratoconus. Pachymetry was measured at the incision site, and the incision was made at 66% of the corneal thickness. A thicker ring segment was typically placed inferiorly, and a thinner segment was placed superiorly on the basis of a refractive nomogram. MAIN OUTCOME MEASURES: Differences between preoperative and postoperative uncorrected visual acuity, best spectacle-corrected acuity, and spherical equivalent. Changes in irregular astigmatism were evaluated with the inferior-superior value from comeotopographic maps, and differences in refractive cylinder groups were studied. RESULTS: Preoperative mean best-corrected logarithm of the minimum angle of resolution (LogMAR) visual acuity was 0.41 (20/50 - 1) (standard deviation [SD], +/-0.48), which improved to a postoperative mean of 0.24 (20/32 - 2) (SD, +/-0.31) (two lines of improvement). Preoperative mean uncorrected LogMAR visual acuity was 1.05 (20/200 - 2 1) (SD, +/-0.48), which improved to a mean of 0.61 (20/80-) (SD, +/-0.52) (four lines of improvement) at postoperative follow-up. Preoperative mean best-corrected LogMAR acuity in the corneal scarring group was 0.96 (20/200 + 2) (SD, +/-0.72), which improved to a mean of 0.54 (SD, +/-0.43) (20/63 - 2) (five lines of improvement). Uncorrected mean LogMAR acuity in the eyes with corneal scarring was 1.42 (20/400 - 4) (SD, +/-0.27), which improved to a mean of 1.03 (20/200 - 1) (SD, +/-73) (three lines of improvement). The mean spherical equivalent before surgery was -3.89 diopters (D) (SD, +/-5.16), which was reduced to a mean of -1.46 D (+/-4.11) at the postoperative follow-up. CONCLUSIONS: Asymmetric Intacs implantation can improve both uncorrected and best spectacle-corrected visual acuity and can reduce irregular astigmatism in corneas with and without corneal scarring. PMID- 12750110 TI - Ischemic optic neuropathy as the first manifestation of elevated cholesterol levels in young patients. AB - PURPOSE: To investigate the relationship between idiopathic nonarteritic ischemic optic neuropathy (NAION) and serum lipid levels in patients /= 240 mg/dl) with NAION was 3.3 (95% confidence interval, 1.4-7.8), and the likelihood increased when the comparison was restricted to nondiabetic patients. Diabetes mellitus was more common in cases than controls (P = 0.027), but systemic hypertension was not significantly different (P = 0.63). No patient (0 of 24) had a magnetic resonance imaging study consistent with optic neuritis or central nervous system demyelination. Visual improvement was uncommon. CONCLUSIONS: This study demonstrates that hypercholesterolemia is associated with NAION in younger patients. NAION may be the first manifestation of a lipid disorder, a previously unrecognized syndrome. These patients have experienced a focal, microvascular central nervous system ischemic event at a relatively young age. Aggressive treatment of lipid abnormalities in these patients may be warranted. PMID- 12750112 TI - Bilateral choroidal folds and optic neuropathy: a variant of the crowded disk syndrome? AB - PURPOSE: To describe the clinical features of the syndrome that includes bilateral choroidal folds and optic neuropathy and to propose a novel etiology to explain this presentation. DESIGN: Observational small case series. PARTICIPANTS: Three patients. METHODS: Three patients received an extensive clinical work-up including complete ocular examination, A and B scan ultrasonography, Humphrey visual field analysis, and imaging of the brain and orbit. Neurological consultation was obtained in each patient two of whom underwent lumbar puncture. The findings of this detailed investigation are reported. RESULTS: Each patient demonstrated bilateral choroidal folds with optic disk congestion and leakage in one eye and optic atrophy in the fellow eye. Extensive ocular and extraocular investigation failed to uncover a specific etiology. Pseudotumor cerebri, a well documented cause of papilledema and choroidal folds, was ruled out in each case. Each patient did demonstrate variable degrees of hyperopia and shortened axial lengths. CONCLUSIONS: The syndrome of bilateral choroidal folds and optic neuropathy may have various etiologies. Pseudotumor cerebri needs to be definitively ruled out. Normal neuro-ophthalmological investigation including lumbar puncture may indicate an alternative cause. Idiopathic acquired hyperopia in middle-aged patients who are hyperopic with shortened axial lengths may be associated with choroidal folds and a constricted scleral canal causing optic disk congestion and complicated by nonarteritic anterior ischemic optic neuropathy due to a crowded disk. PMID- 12750113 TI - Acute concomitant esotropia of adulthood. AB - PURPOSE: To identify the characteristics of adult patients who develop acute concomitant esotropia of adulthood. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Ten patients were included in this study. INTERVENTION: The charts of all adults with acute-onset concomitant esotropia who were examined at our institute between 1990 and 1997 were reviewed, and those who had developed the syndrome when they were older than the age of 16 years were included in this study. All participants underwent a complete ocular and physical examination, including brain and orbital computed tomography. MAIN OUTCOME MEASURES: Angle of esotropia, measured by the prism and cover test. RESULTS: The mean age +/- standard deviation at the time of the ocular and physical examination was 38 +/- 18.6 years (range, 18-70 years). The mean myopic error was -4.1 +/- 3.2 diopters (range, +2.0 to -8.5 diopters). Nine of the 10 patients were myopic. The mean angle of esotropia was 33.8 +/- 14.7 prism diopters (range, 18-60 prism diopters). The mean period of follow-up was 2.2 +/- 1.0 years (range, 1-4 years). After surgery, all patients were orthophoric or minimally esophoric, and in all of them, stereoacuity (measured by the Titmus stereofly test) was 40 arc seconds. CONCLUSIONS: In a well-defined group of adult patients with acute-onset concomitant esotropia, almost all were myopic, and all regained normal stereopsis after surgery. Acute concomitant esotropia of adulthood should probably be classified as a distinct subgroup of acute-onset esotropia. PMID- 12750114 TI - Multifocal nodular episcleritis and scleritis with undiagnosed Hodgkin's lymphoma. AB - PURPOSE: To report the case of a patient with undiagnosed Hodgkin's lymphoma who presented with coexistent unilateral nodular episcleritis and scleritis. DESIGN: Interventional case report and literature review METHODS: Review of clinical history, laboratory findings, histology of episcleral and cervical lymph node biopsies, and follow-up. RESULTS: A 20-year-old female presented with a 5-month history of redness and pain in her left eye, with associated symptoms of dyspnea, malaise, and fever. The patient was found to have multifocal nodular episcleritis and scleritis that was not responsive to topical steroids or systemic nonsteroidal anti-inflammatory treatment. Laboratory tests subsequently revealed evidence of systemic inflammation, and radiologic studies showed extensive mediastinal and cervical adenopathy. A cervical lymph node biopsy showed Reed Sternberg cells and a chronic lymphocytic infiltrate consistent with nodular sclerosing Hodgkin's lymphoma. Histopathologic analysis of an episcleral nodule revealed a necrotizing granuloma with vasculitis. Systemic chemotherapy was instituted for the Hodgkin's disease; this therapy abolished the nodular scleritis. CONCLUSIONS: This case raises the possibility of concurrent undiagnosed systemic vasculitis with only an ocular manifestation with Hodgkin's lymphoma, either as a coincidence or as a paraneoplastic syndrome. Moreover, it emphasizes the important role of tissue biopsy in establishing diagnosis and directing treatment. PMID- 12750115 TI - Mycophenolate mofetil as an immunomodulatory agent in the treatment of chronic ocular inflammatory disorders. AB - PURPOSE: To evaluate the outcomes of patients with chronic ocular inflammatory disease treated with mycophenolate mofetil as an immunosuppressive and steroid sparing agent. DESIGN: Retrospective noncomparative interventional case series. PARTICIPANTS: All patients with ocular inflammatory disease treated with mycophenolate mofetil at a single institution between 1998 and 2001. METHODS: Charts of patients seen on the Ocular Immunology and Uveitis Service at the Massachusetts Eye and Ear Infirmary were reviewed. Patients with chronic ocular inflammatory disease were included in the study. MAIN OUTCOME MEASURES: Control of inflammation, steroid-sparing effect, visual acuity, and adverse reactions were measured. RESULTS: A total of 54 patients were evaluated. Control of ocular inflammation with mycophenolate mofetil as monotherapy was achieved in 35 patients (65%) and in 67 eyes (62%), and a steroid-sparing effect was achieved in 29 (54%) patients. Visual acuity was maintained or improved in 51 patients (94%) and in 97 eyes (90%). Side effects requiring discontinuation of medication occurred in 10 patients (18%). There was neither long-term morbidity nor mortality due to mycophenolate mofetil. CONCLUSIONS: Mycophenolate mofetil is effective in the treatment of patients with steroid-dependent or -resistant chronic ocular inflammatory disorders that fail to respond to conventional steroid treatment. It is a safe and effective steroid-sparing immunomodulatory agent and can be considered an important addition to our armamentarium in the care of patients with ocular inflammatory disease. PMID- 12750117 TI - Lysosomal enzymes are released from cultured human macrophages, hydrolyze LDL in vitro, and are present extracellularly in human atherosclerotic lesions. AB - OBJECTIVE: Human atherosclerotic lesions have been shown to contain lipid droplets and vesicles resembling those of in vitro enzymatically modified LDL. However, little is known about the hydrolytic enzymes in the arterial intima that induce fusion of LDL particles and so produce lipid droplets or that induce foam cell formation. METHODS AND RESULTS: Human coronary atherosclerotic lesions obtained at surgery and at autopsy were stained for lysosomal acid lipase and cathepsin D. The extracellular areas of macrophage-rich intimal regions of the atherosclerotic lesions stained positively for both cathepsin D and lysosomal acid lipase, whereas normal arteries were negative. When monocyte-derived macrophages were incubated with opsonized zymosan to stimulate the release of lysosomal enzymes from the cells and LDL was incubated with the macrophage conditioned media, the apolipoprotein B-100, cholesteryl esters, and triacylglycerols of LDL were hydrolyzed. These hydrolytic modifications rendered the LDL particles unstable and induced their fusion. Cultured macrophages and smooth muscle cells took up the hydrolase-modified LDL particles avidly and were transformed into foam cells. CONCLUSIONS: Our in vivo and in vitro results suggest that lysosomal enzymes released from macrophages may induce hydrolytic modification of LDL and foam cell formation in the human arterial intima. PMID- 12750116 TI - Contribution of von Willebrand factor to thrombus formation on neointima of rabbit stenotic iliac artery under high blood-flow velocity. AB - OBJECTIVE: It has become clear that von Willebrand factor (vWF) plays important roles in platelet adhesion and aggregation under high blood-flow velocity conditions observed in stenotic atherosclerotic arteries. However, its roles in thrombus formation in vivo on diseased arteries have not been fully understood. We examined the contribution of vWF to thrombus formation and subsequent intimal growth by using a repeated balloon-injury model in rabbits. METHODS AND RESULTS: Rabbit iliac arteries 4 weeks after a first balloon injury showed 37% luminal stenosis by neointimal growth, and blood velocity increased by 2.1 times compared with that of uninjured arteries. The second balloon injury induced fibrin-rich thrombus formation on the injured neointima. Intravenous administration of a monoclonal antibody against vWF (AJW200, 1.0 mg/kg body weight) remarkably prevented botrocetin-induced platelet aggregation ex vivo for 2 days; moreover, thrombus formation, cell proliferation, and subsequent neointimal growth were significantly reduced at 30 minutes, 5 days, and 4 weeks, respectively, after the second balloon injury. CONCLUSIONS: These results indicate that vWF plays a potent role in fibrin-rich thrombus formation on the neointima under high blood flow velocity conditions. Inhibition of plasma vWF activity might be effective for the reduction of thrombus formation and/or subsequent neointimal development after coronary interventions. PMID- 12750118 TI - Small, dense LDL and elevated apolipoprotein B are the common characteristics for the three major lipid phenotypes of familial combined hyperlipidemia. AB - OBJECTIVE: Familial combined hyperlipidemia (FCHL) is associated with variable lipid and lipoprotein phenotypes arbitrarily defined as type IIa, IIb, and IV based on plasma total cholesterol and triglyceride levels. This study sought to characterize consistent lipoprotein and lipid abnormalities across the 3 lipoprotein phenotypes in 62 patients with documented FCHL (IIa [n=14], IIb [n=19], and IV [n=29]) and 44 healthy individuals. METHODS AND RESULTS: The lipoprotein cholesterol distribution was determined over 38 fractions obtained by density gradient ultracentrifugation. As expected, FCHL patients with hypertriglyceridemia (IIb and IV) had higher cholesterol levels in VLDL than IIa, whereas IIa showed higher cholesterol in the big, buoyant LDL and in HDL. LDL cholesterol was higher in IIb than IV; most of the increase in LDL cholesterol was associated with big, buoyant LDL rather than small, dense LDL (sdLDL). The differences in lipoproteins between phenotypes were attributable to changes in VLDL and big, buoyant LDL levels. Comparison of the FCHL patients with healthy individuals showed a significant elevation in plasma apolipoprotein B levels and sdLDL in all 3 FCHL phenotypes. CONCLUSIONS: Although triglyceride and cholesterol levels are variable by lipoprotein phenotype, sdLDL and elevated plasma apolipoprotein B levels are consistent characteristics of FCHL shared by the 3 different lipoprotein phenotypes. PMID- 12750119 TI - Inhibition of purified factor Xa amidolytic activity may not be predictive of inhibition of in vivo thrombosis: implications for identification of therapeutically active inhibitors. AB - OBJECTIVE: In this study we test the hypothesis that blood/plasma-based prothrombinase assays, rather than inhibition of purified factor Xa (fXa), are predictive of in vivo antithrombotic activity. METHODS AND RESULTS: Six fXa inhibitors with equivalent nanomolar Ki were studied in thrombin generation assays using human plasma/blood and endogenous macromolecular substrate. In all assays, benzamidine inhibitors were more potent (100 to 800 nmol/L) than the aminoisoquinolines (5 to 58 micromol/L) or neutral inhibitors (3 to 10 micromol/L). A similar rank order of compound inhibition was also seen in purified prothrombinase assays as well as in a rabbit model of deep vein thrombosis. CONCLUSIONS: Assays using prothrombinase with protein substrates are better predictors of in vivo efficacy than fXa Ki using amidolytic substrates. PMID- 12750120 TI - Plasma PAI-1 levels are more strongly related to liver steatosis than to adipose tissue accumulation. AB - OBJECTIVE: Because obesity and insulin resistance (IR) are strongly associated with liver steatosis (LS), we investigated the relation between the degree of LS and plasminogen activator inhibitor-1 (PAI-1) in ob/ob mice, in C57/BL6 mice with alcoholic LS, and in severely obese humans. METHODS AND RESULTS: In both mouse models, plasma PAI-1 levels were associated with PAI-1 expression in the liver and with the degree of LS. Liver PAI-1 antigen was associated with the tumor necrosis factor receptor-II (TNFRII) antigen, whereas association with TNF antigen content was found in ob/ob mice only. No significant correlation between plasma PAI-1 and PAI-1 expression in adipose tissue of ob/ob mice was observed. Furthermore, the relation between plasma PAI-1 levels and body weight was positive in ob/ob mice but negative in C57/BL6 mice (both P<0.001). In humans, PAI-1 levels were correlated with the degree of LS, and 26% of plasma PAI-1 activity was independently explained by LS and serum insulin levels. CONCLUSIONS: Plasma PAI-1 levels are more closely related to fat accumulation and PAI-1 expression in the liver than in adipose tissue. In steatotic liver, PAI-1 antigen content is associated with those of TNF and TNFRII. Therefore, we suggest that TNF pathway dysregulation in LS could be involved in increased plasma PAI-1 in obesity with IR. PMID- 12750122 TI - Conflict of interest? PMID- 12750121 TI - Mechanisms of renal structural alterations in combined hypercholesterolemia and renal artery stenosis. AB - OBJECTIVE: Atherosclerotic renovascular disease (ARVD) aggravates renal scarring more than other causes of renal artery stenosis (RAS), but the underlying pathogenic mechanisms of this potential profibrotic effect remain unclear. We tested the hypothesis that coexistence of atherosclerosis and RAS interferes with renal tissue remodeling. METHODS AND RESULTS: Single-kidney hemodynamics and function were quantified in vivo with electron-beam computed tomography in 3 groups of pigs (n=7 each): normal pigs, pigs 12 weeks after induction of unilateral RAS (RAS group), and pigs with similar-degree RAS fed a 12-week 2% hypercholesterolemic diet (HC+RAS, simulating early ARVD). Kidneys were studied ex vivo by Western blotting and immunohistochemistry. Renal volume, renal blood flow, and glomerular filtration rate were similarly decreased in RAS and HC+RAS ischemic kidneys, accompanied by similar increased expression of profibrotic factors like transforming growth factor-beta, tissue inhibitor of metalloproteinase-1, and plasminogen activator inhibitor-1. Nevertheless, HC+RAS kidneys showed increased intrarenal fibrosis compared with RAS-only kidneys. Furthermore, expression of nuclear factor-kappaB was increased, expression of extracellular (matrix metalloproteinase-2) and intracellular (ubiquitin) protein degradation systems was decreased, and apoptosis was blunted. CONCLUSIONS: Diet induced HC superimposed on RAS accelerates the development of fibrosis in the stenotic kidney by amplifying profibrotic mechanisms and disrupting tissue remodeling. These alterations might contribute to renal disease progression in ARVD and might account for the increased propensity for end-stage renal disease. PMID- 12750123 TI - Biomechanical evaluation of a new ulnar collateral ligament reconstruction technique with interference screw fixation. AB - BACKGROUND: Techniques for ulnar collateral ligament reconstruction have evolved. HYPOTHESIS: Ulnar collateral ligament reconstruction with interference screw fixation restores elbow kinematics and failure strength to that of the native ligament. STUDY DESIGN: Controlled laboratory study. METHODS: Of 10 matched pairs of cadaveric elbows, one underwent kinematic testing under conditions of an intact, released, and reconstructed ligament. Single 5-mm diameter bone tunnels were created at the isometric anatomic insertion sites on the medial epicondyle and sublime tubercle. Graft fixation was achieved with 5 x 15 mm soft tissue interference screws. The reconstructed and contralateral intact elbows were then tested to failure. RESULTS: Average stiffness for intact elbows (42.81 +/- 11.6 N/mm) was significantly greater than for reconstructed elbows (20.28 +/- 12.5 N/mm). Ultimate moment for intact elbows (34.0 +/- 6.9 N.m) was not significantly different from reconstructed elbows (30.6 +/- 19.2 N.m). Release of the ulnar collateral ligament caused a significant increase in valgus instability. Reconstruction restored valgus stability to near that of the intact elbow. CONCLUSIONS: With this reconstruction method, failure strength was comparable with that of the native ligament and physiologic elbow kinematics were reliably restored. CLINICAL RELEVANCE: This technique returns elbow kinematics to near normal, with less soft tissue dissection and risk of ulnar nerve injury and ease of graft insertion, tensioning, and fixation. PMID- 12750124 TI - Multistranded hamstring tendon graft fixation with a central four-quadrant or a standard tibial interference screw for anterior cruciate ligament reconstruction. AB - BACKGROUND: Tibial fixation of hamstring tendon grafts has been the weak link in anterior cruciate ligament reconstruction. HYPOTHESIS: Use of a central four quadrant sleeve and screw provides superior fixation when compared with standard interference screw fixation. STUDY DESIGN: Controlled laboratory study. METHODS: In eight pairs of cadaveric knees each anterior cruciate ligament was reconstructed using either an interference screw or a central sleeve and screw on the tibial side. The specimens were then subjected to cyclic loading followed by a load-to-failure test. RESULTS: The load required to cause 1 and 2 mm of graft laxity, defined as the separation of the femur and the tibia at the points of graft fixation, was significantly greater with the sleeve and screw than with the interference screw (at 2 mm: sleeve and screw, 216.1 +/- 30.1 N; interference screw, 167.0 +/- 33.2 N). The force at initial slippage for each of the graft strands was significantly higher with use of the central sleeve and screw. CONCLUSIONS: The four-quadrant sleeve and screw device may provide greater surface area for healing of hamstring tendon grafts and allow equal tensioning of graft strands before fixation. These factors are associated with increased strength of fixation and reduced laxity of the graft after cyclic loading. CLINICAL RELEVANCE: Use of the central four-quadrant sleeve and screw system offers increased strength of fixation in anterior cruciate ligament reconstruction with hamstring tendon graft. PMID- 12750125 TI - Determination of time of biologic fixation after anterior cruciate ligament reconstruction with hamstring tendons. AB - BACKGROUND: It has been unclear when a hamstring tendon graft becomes biologically fixed in the bone tunnel after anterior cruciate ligament reconstruction. HYPOTHESIS: Postoperative biomechanical testing and magnetic resonance images can indicate biologic fixation of the graft in the femoral bone tunnel. STUDY DESIGN: Prospective cohort study. METHODS: Sixty-four patients were evaluated by serial biomechanical testing, magnetic resonance imaging, and second look arthroscopy 2 years after surgery. Biologic fixation of the graft was confirmed radiographically by injecting a contrast medium into the femoral bone tunnel. RESULTS: Forty-two stable knees with graft fixation maintained a high stiffness (120% of normal) and showed low signal intensity in an early postoperative magnetic resonance image (12 +/- 8 months). Fourteen stable knees without graft fixation had gradually increased anterior displacement with nearly normal stiffness and high signal intensity. Five unstable knees with graft fixation retained low stiffness (70%) and showed late low signal intensity at 20 +/- 9 months. Three unstable knees without biologic fixation had rapidly increased anterior displacement, with half the stiffness of a normal knee. CONCLUSION: Postoperative low stiffness and high signal intensity might indicate late biologic graft fixation, predicting a possibility of postoperative anterior knee instability. PMID- 12750126 TI - Compaction drilling does not increase the initial fixation strength of the hamstring tendon graft in anterior cruciate ligament reconstruction in a cadaver model. AB - BACKGROUND: Compaction of the bone tunnel walls has been proposed to increase the fixation strength of soft tissue grafts fixed with an interference screw in anterior cruciate ligament reconstructions. HYPOTHESIS: Compaction drilling does not increase the initial fixation strength of the hamstring tendon graft in comparison with conventional extraction drilling. STUDY DESIGN: Randomized experimental study. METHODS: Initial fixation strength of quadrupled hamstring tendon grafts fixed with bioabsorbable interference screws was assessed in 22 pairs of human cadaveric tibiae. Bone tunnels were drilled with either a compaction drill or a conventional extraction drill. Specimens underwent a cyclic loading test and the surviving specimens were then loaded to failure in a single cycle load-to-failure test. Trabecular bone mineral density at the site corresponding to the actual site of the tibial bone tunnel was determined by using peripheral quantitative computed tomography. RESULTS: During the cyclic loading test, no significant stiffness or displacement differences were observed between the two drilling techniques. Three specimens failed in the compaction drilling group, whereas there were no failures in the extraction-drilling group. In the subsequent single-cycle load-to-failure test, no significant differences between the two drilling techniques were found with regard to displacement at yield load, stiffness, or mode of failure. There was no significant difference in trabecular bone mineral density between the two groups. CONCLUSIONS: Compaction drilling does not increase the initial fixation strength of the hamstring tendon graft compared with conventional extraction drilling. PMID- 12750127 TI - Repair of pars interarticularis defect by segmental wire fixation in young athletes with spondylolysis. AB - BACKGROUND: Although segmental wire fixation has been successful in the treatment of nonathletes with spondylolysis, no information exists on the results of this type of surgery in athletes. PURPOSE: To evaluate the outcome of surgical repair of pars interarticularis defect by segmental wire fixation in young athletes with lumbar spondylolysis. METHODS: Between 1993 and 2000, 20 athletes (6 women and 14 men; average age, 23.7) with lumbar spondylolysis were treated surgically with this technique. They were actively engaged in sports such as baseball, tennis, and golf. Nineteen athletes had one level of spondylolysis and one athlete had two levels. The level of spondylolysis was L4 in 2 athletes and L5 in 19. The average follow-up period was 3.5 years (range, 1.3 to 8.6). Surgical outcome was evaluated by radiographic examination, the Japanese Orthopaedic Association score, preoperative and postoperative sports activity levels and intensities, and the presence of complications. RESULTS: Bony fusion at the site of spondylolysis was obtained in all cases, and the Japanese Orthopaedic Association score was increased significantly after surgery (preoperatively, 21.2 +/- 3.9; postoperatively, maximum 27.7 +/- 1.0; recovery rate, 80.4%). All of the patients returned to their sports activities, although at varying degrees. No severe complications were noted. CONCLUSION: We recommend this technique in cases of lumbar spondylolysis in athletes who hope to resume their sports activities. PMID- 12750128 TI - Injuries in a modern dance company: effect of comprehensive management on injury incidence and time loss. AB - BACKGROUND: Professional dancers experience high rates of musculoskeletal injuries. OBJECTIVE: To analyze the effect of comprehensive management (case management and intervention) on injury incidence, time loss, and patterns of musculoskeletal injury in a modern dance organization. STUDY DESIGN: Retrospective/prospective cohort study. METHODS: Injury data were analyzed over a 5-year period, 2 years without intervention and 3 years with intervention, in a modern dance organization (42 dancers). The number of workers' compensation cases and number of dance days missed because of injury were compared across a 5-year period in a factorial design. RESULTS: Comprehensive management significantly reduced the annual number of new workers' compensation cases from a high of 81% to a low of 17% and decreased the number of days lost from work by 60%. The majority of new injuries occurred in younger dancers before the implementation of this program. Most injuries involved overuse of the lower extremity, similar to patterns reported in ballet companies. Benefits of comprehensive management included early and effective management of overuse problems before they became serious injuries and triage to prevent overutilization of medical services. CONCLUSIONS: This comprehensive management program effectively decreased the incidence of new cases and lost time. Both dancers and management strongly support its continuance. PMID- 12750129 TI - Biomechanical comparison of the FasT-Fix meniscal repair suture system with vertical mattress sutures and meniscus arrows. AB - BACKGROUND: A meniscal repair technique that combines the strength of vertical mattress sutures and the decreased tissue morbidity of an all-inside technique would be advantageous. HYPOTHESIS: The FasT-Fix Meniscal Repair Suture System will provide load at failure, stiffness, and displacement equivalent to that of vertical mattress sutures and superior to that of Meniscus Arrows. STUDY DESIGN: In vitro biomechanical study. METHODS: After repair of a 2-cm vertical longitudinal medial meniscal lesion, three groups of six human cadaveric knees were biomechanically tested in a random order on a servohydraulic device, and three groups of five specimens underwent cyclic loading. RESULTS: Specimens repaired with Meniscus Arrows had reduced load at failure, stiffness, and displacement, but there were no differences between the FasT-Fix and vertical mattress suture methods. During cyclic loading, specimens repaired with two Meniscus Arrows failed before test completion, whereas specimens repaired with two vertical mattress sutures (6.0 +/- 3.7 mm) or with two FasT-Fix implants (5.1 +/- 1.4 mm) maintained fixation with comparable displacements. CONCLUSIONS: The FasT-Fix provided load at failure, stiffness, and displacement comparable with that of vertical mattress sutures. CLINICAL RELEVANCE: The results suggest that the FasT-Fix may be preferable to Meniscus Arrows for meniscal repair with minimal associated tissue morbidity. PMID- 12750130 TI - Rates and risks of injury during intercollegiate basketball. AB - BACKGROUND: Previous studies of basketball injury have not been able to assess injury incidence and risk. PURPOSE: To determine rates and risks of injury in Canadian intercollegiate basketball. STUDY DESIGN: Prospective cohort study. METHODS: Standardized data were collected with a validated instrument from 98.1% of the 318 athletes on the eight men's basketball teams in the Canada West Division of the Canadian Intercollegiate Athletic Union. RESULTS: A total of 142 athletes sustained 215 injuries (44.7% of players injured) over the 2-year study period. The greatest number of injuries resulting in more than seven sessions of time loss involved the knee, whereas the most common injuries causing fewer than seven sessions of time loss involved the ankle. The most common mechanism of injury was contact with another player, especially in the "key." Injuries occurred 3.7 times more often in games than during practice. Centers had the highest rate of injury, followed by guards, and then forwards. The relative risk of reinjury was significantly increased by previous injuries to the elbow, shoulder, knee, hand, lower spine or pelvis, and by concussions. CONCLUSIONS: Risk factors for injury were previous injury, games as opposed to practice, player position, player contact, and court location. PMID- 12750131 TI - The effects of radiofrequency energy treatment on chondrocytes and matrix of fibrillated articular cartilage. AB - BACKGROUND: There is no consensus regarding the safety of radiofrequency energy treatment for chondroplasty. HYPOTHESIS: Use of a radiofrequency thermal probe will produce a dose-dependent detrimental effect on chondrocytes in a setting mimicking the arthroscopic procedure. STUDY DESIGN: Controlled laboratory study. METHODS: Paired patellae from 11 horses were fibrillated; one served as the control while the contralateral patella was treated with 20, 40, and 60 watts of radiofrequency energy for 4 minutes. Proteoglycan synthesis, proteoglycan degradation, and cell viability were measured. Explant histologic sections were scored for cellular characteristics, metachromatic matrix staining intensity, and tissue architecture and were quantitatively analyzed for cell death. RESULTS: Mean peak surface articular cartilage temperatures were 35.47 degrees C (20 watts), 40.76 degrees C (40 watts), and 44.81 degrees C (60 watts). Treatment at all three settings significantly decreased proteoglycan synthesis of the chondrocytes. Proteoglycan degradation increased with increased power setting. Treatment at 40 and 60 watts significantly decreased cell viability to 81% and 73%, respectively. Significant histologic changes occurred in the superficial cartilage zone with 60-watt application. CONCLUSION: Use of radiofrequency energy on mechanically fibrillated articular cartilage induced a dose-dependent detrimental effect on chondrocytes and matrix metabolism. CLINICAL RELEVANCE: Cautious use of radiofrequency energy for treatment of articular cartilage is recommended until long-term effects are evaluated. PMID- 12750132 TI - Recovery of chondrocyte metabolic activity after thermal exposure. AB - BACKGROUND: The relationship between temperature elevation and thermal exposure time during thermal chondroplasty has implications for cell viability and subsequent articular cartilage function. PURPOSE: To characterize cartilage metabolic changes after exposure to thermal stress and to determine whether changes seen acutely are reversible. STUDY DESIGN: Controlled laboratory study. METHODS: Human cartilage was exposed to a 45 degrees, 50 degrees, or 55 degrees C bath for up to 3 minutes. Untreated control specimens were analyzed with each group. Viability and metabolic capability of treated and untreated specimens were evaluated immediately or 1 week after thermal stress by using methylthiotetrazole conversion, (3)H-serine incorporation into protein, and (35)S-sulfate incorporation into newly synthesized proteoglycan. RESULTS: Nonarthritic and arthritic articular cartilage metabolic activity declined with increasing thermal exposure. Articular cartilage displayed a recovery from thermal stress after exposure to the 50 degrees C but not the 55 degrees C bath. Arthritic cartilage displayed increased sensitivity with higher temperatures. CONCLUSIONS: Understanding of the increased sensitivity to thermal stress of arthritic articular cartilage may be helpful in thermally based treatments. CLINICAL RELEVANCE: Further correlation with the temperatures attained during thermal chondroplasty will be necessary to confirm the clinical relevance of these in vitro observations to the use of radiofrequency energy devices to treat partial thickness chondral lesions. PMID- 12750133 TI - Dynamic stabilizing function of the deltoid muscle in shoulders with anterior instability. AB - BACKGROUND: The stabilizing role of the deltoid muscle has not been extensively studied. PURPOSE: To study the contribution of the deltoid muscle to anterior stability of the shoulder. STUDY DESIGN: Controlled laboratory study. METHODS: We used nine fresh cadaveric shoulders with the arm at 90 degrees of abduction and 90 degrees of external rotation. The position of the humeral head was monitored by an electromagnetic tracking device with 0 and 1.5 kg of anterior translation force; with 0, 1, 3, and 5 kg of force applied to each of the anterior, middle, and posterior portions of the deltoid muscle; and with the capsule intact, vented, and with a simulated Bankart lesion. RESULTS: With the capsule intact, anterior displacement was significantly reduced by application of load to the middle deltoid muscle. After the capsule was vented, load application to the anterior, middle, or posterior deltoid muscle significantly reduced anterior displacement. With a simulated Bankart lesion, effects of muscle loading were most apparent: anterior displacement was significantly reduced with loading of each muscle portion. CONCLUSION: The deltoid muscle is an anterior stabilizer of the glenohumeral joint with the arm in abduction and external rotation. CLINICAL RELEVANCE: The stabilizing function of the deltoid muscle takes on more importance as the shoulder becomes unstable. PMID- 12750134 TI - Tibial eminence fractures in children: prevalence of meniscal entrapment. AB - BACKGROUND: Meniscal entrapment under a displaced tibial eminence fragment may be a rationale for arthroscopic or open reduction in type 2 and 3 tibial eminence fractures. PURPOSE: To determine the prevalence of meniscal entrapment in children with type 2 and 3 tibial eminence fractures. STUDY DESIGN: Case series. METHODS: Records of a consecutive series of 80 skeletally immature patients (mean age, 11.6 years; range, 5 to 16) who underwent arthroscopic (71), open (5), or combined arthroscopic and open (4) reduction and internal fixation of type 3 tibial eminence fractures (57) or type 2 fractures that did not reduce in extension (23) from 1993 to 2001 were reviewed. RESULTS: Entrapment of the anterior horn of the medial meniscus (36), intermeniscal ligament (6), or anterior horn of the lateral meniscus (1) was seen in 26% (6 of 23) of type 2 fractures and 65% (37 of 57) of type 3 fractures. An associated meniscal tear was seen in 3.8% of patients (3 of 80). CONCLUSIONS: Meniscal entrapment is common in patients with type 2 and 3 tibial eminence fractures. Arthroscopic or open reduction should be considered for type 3 fractures and for type 2 fractures that do not reduce in extension to remove the incarcerated meniscus, allowing for anatomic reduction. PMID- 12750135 TI - Performance characteristics of volleyball players with patellar tendinopathy. AB - BACKGROUND: Patellar tendinopathy is assumed to result from chronic tendon overload. There may be a relationship between tendon pain and jumping ability. HYPOTHESIS: There is no difference in performance characteristics between volleyball players with patellar tendinopathy and those without. STUDY DESIGN: Prospective cohort study. METHOD: We examined the performance of the leg extensor apparatus in high-level male volleyball players with patellar tendinopathy (N = 24) compared with a control group (N = 23) without knee symptoms. The testing program consisted of different jump tests with and without added load, and a composite jump score was calculated to reflect overall performance. RESULTS: The groups were similar in age, height, and playing experience, but the patellar tendinopathy group did more specific strength training and had greater body weight. They scored significantly higher than the control group on the composite jump score (50.3 versus 39.2), and significant differences were also observed for work done in the drop-jump and average force and power in the standing jumps with half- and full-body weight loads. CONCLUSIONS: Greater body weight, more weight training, and better jumping performance may increase susceptibility to patellar tendinopathy in volleyball players. PMID- 12750136 TI - Treatment of postoperative anterior cruciate ligament infections with graft removal and early reimplantation. AB - BACKGROUND: Septic arthritis after arthroscopic anterior cruciate ligament reconstruction is rare, and the most appropriate treatment is unclear. Current recommendations are that, if the graft is removed, reimplantation should be delayed for 6 to 9 months. HYPOTHESIS: Early removal of the graft with appropriate infection management followed by early reimplantation can lead to good results. STUDY DESIGN: Uncontrolled retrospective review. METHODS: Records of all patients who developed postoperative infection after anterior cruciate ligament reconstruction were reviewed. Four patients had early graft removal and appropriate infection management including 6 weeks of intravenous antibiotics followed by anterior cruciate ligament graft reimplantation within 6 weeks of completion of antibiotic therapy. RESULTS: Follow-up at an average of 21 months (range, 14 to 31) showed that the patients treated with early reimplantation had full symmetric knee range of motion and no effusion. The average modified Lysholm score was 92.5. Radiographs demonstrated no joint-space narrowing or osteophyte formation. The 30-pound KT-1000 arthrometer side-to-side difference averaged 3 mm. CONCLUSION: Graft removal after confirmed anterior cruciate ligament graft infection and intravenous antibiotic administration followed by early graft reimplantation can give excellent results. PMID- 12750137 TI - Effect of drain use in the early postoperative period after arthroscopically assisted anterior cruciate ligament reconstruction with bone-patellar tendon-bone graft. AB - BACKGROUND: Little data exist on the effect of routine use of postoperative drainage after arthroscopic anterior cruciate ligament reconstruction, although clinical studies of other procedures have not shown benefit to this practice. HYPOTHESIS: Use of a postoperative drain will not result in decreased suprapatellar girth, increased range of motion, and decreased pain compared with nonuse. STUDY DESIGN: Prospective randomized clinical trial. METHODS: Twenty-one patients undergoing arthroscopically assisted bone-patellar tendon-bone anterior cruciate ligament reconstruction were randomly assigned to receive a drain for 24 hours (12 patients) or no drain (9 patients). Data for comparison of groups were collected daily through postoperative day 7. RESULTS: Pain scores on a visual analog scale demonstrated the same improving trend over time for both treatment and control groups; however, the treatment group had significantly higher average pain scores, except on day 7. Differences in suprapatellar girth, flexion, and extension were not found to be statistically significant between groups. CONCLUSION: Use of a drain after arthroscopically assisted anterior cruciate ligament reconstruction provided no benefit in terms of range of motion, effusion, or pain in the early postoperative period. PMID- 12750138 TI - Shock wave therapy for calcific tendinitis of the shoulder: a prospective clinical study with two-year follow-up. AB - BACKGROUND: Shock wave therapy is a new modality that has shown efficacy in the treatment of various orthopaedic disorders. PURPOSE: To determine the effectiveness, at 2- to 3-year follow-up, of shock wave therapy for calcific tendinitis of the shoulder. STUDY DESIGN: Prospective clinical study. METHODS: Thirty-seven patients (39 shoulders) with calcific shoulder tendinitis were treated with shock wave therapy (1000 impulses at 14 kV) and observed for 24 to 30 months. The control group, which underwent sham treatment with a dummy electrode, consisted of 6 patients (6 shoulders) with an average follow-up of 6 months. Evaluation included use of the 100-point Constant score system and shoulder radiographs. RESULTS: The overall results in the study group were 60.6% excellent (20 of 33 shoulders), 30.3% good (10), 3.0% fair (1), and 6.1% poor (2), and those of the control group were 16.7% fair (1 of 6 shoulders) and 83.3% poor (5). The symptom recurrence rate in the study group was 6.5%. Dissolution of calcium deposits was complete in 57.6% of the study group, partial in 15.1%, and unchanged in 27.3%. Fragmentation was seen in 16.7% of the control group patients; in 83.3% deposits were unchanged. No recurrence of calcium deposits was observed during the 2 years that the study group was followed. CONCLUSIONS: Shock wave therapy is a safe and effective noninvasive treatment for patients with calcific tendinitis of the shoulder. PMID- 12750139 TI - Assessment of shoulder proprioception in the female softball athlete. AB - BACKGROUND: There have been reports of overhand throwing athletes having decreased joint position sense in their dominant shoulder as compared with the nondominant shoulder. Very little research, however, exists concerning joint position sense in the female athlete. HYPOTHESIS: Female softball athletes have decreased joint position sense in their dominant shoulder as compared with their nondominant shoulder. STUDY DESIGN: Factorial design with investigation of multiple independent variables. METHODS: Joint position sense was assessed in 50 female softball players and 50 nonthrowing female athletes by using an inclinometer during four glenohumeral joint motions. Both the dominant and nondominant shoulders were assessed and error scores were calculated to describe joint position sense. Data were collected during the course of a fall semester and analyzed by using a mixed model analysis of variance with repeated measures on the dependent variable (error scores). RESULTS: A significant group by movement interaction was observed, with the softball athletes demonstrating significantly greater external rotation error scores than the nonthrowing athletes. CONCLUSION: We failed to reject the null hypothesis. Increased error scores (less joint position sense) were observed in both arms of subjects in the softball group. CLINICAL RELEVANCE: This study suggests that there is decreased shoulder proprioception in asymptomatic female athletes involved in overyhand throwing sports, which may predispose them to injury. PMID- 12750140 TI - Injuries and overuse syndromes in golf. AB - BACKGROUND: Although golf is becoming more popular, there is a lack of reliable epidemiologic data on golf injuries and overuse syndromes, especially regarding their severity. OBJECTIVE: To perform an epidemiologic study of the variety of different musculoskeletal problems in professional and amateur golfers and to find associations of age, sex, physical stature (body mass index), warm-up routine, and playing level with the occurrence of reported injuries. STUDY DESIGN: Retrospective cohort study. METHODS: We analyzed the injury data from a total of 703 golfers who were randomly selected over two golfing seasons and interviewed with the use of a six-page questionnaire. RESULTS: Overall, 82.6% (N = 526) of reported injuries involved overuse and 17.4% (N = 111) were single trauma events. Professional golfers were injured more often, typically in the back, wrist, and shoulder. Amateurs reported many elbow, back, and shoulder injuries. Severity of reported injuries was minor in 51.5%, moderate in 26.8%, and major in 21.7% of cases. Carrying one's bag proved to be hazardous to the lower back, shoulder, and ankle. Warm-up routines were found to have a positive effect if they were at least 10 minutes long. CONCLUSIONS: Overall, golf may be considered a rather benign activity, if overuse can be avoided. If not, golf can result in serious, chronic musculoskeletal problems. PMID- 12750141 TI - Influence of the femoral tunnel location and angle on the contact pressure in the femoral tunnel in anterior cruciate ligament reconstruction. AB - BACKGROUND: The causes of bone tunnel enlargement after anterior cruciate ligament reconstruction have not been thoroughly investigated. HYPOTHESIS: A malpositioned femoral tunnel and an acute femoral tunnel angle may increase the mechanical stress in the femoral tunnel. STUDY DESIGN: Controlled laboratory study. METHODS: Three femoral tunnels (normal, anterior, and acute) and the tibial tunnel were made in four fresh-frozen cadaveric knees. Aluminum cylinders containing pressure-sensitive conductive rubber sensors at the joint entrance were inserted into the femoral tunnels. Dynamic changes in the contact pressure of the graft in the femoral tunnel were measured. RESULTS: Peak contact pressures and dynamic changes in contact pressure for the normal and anterior femoral tunnels demonstrated no differences. Maximum contact pressure of the graft was observed at the anterior portion with the knee in full extension and at the posterior portion with deep knee flexion. Consistent contact pressure occurred at the anterior aspect of the acute femoral tunnel throughout the range of motion. Mean contact pressure at the anterior region of the acute femoral tunnel was significantly higher than that of the normal femoral tunnel at 60 degrees, 90 degrees, and 120 degrees of knee flexion. CONCLUSIONS: The consistent contact pressure in the anterior aspect of the acute femoral tunnel may erode the anterior portion of the femoral tunnel, resulting in bone tunnel enlargement. CLINICAL RELEVANCE: The femoral tunnel direction in anterior cruciate ligament reconstruction is an important factor in reducing femoral tunnel enlargement. PMID- 12750142 TI - Differences in kinematics and electromyographic activity between men and women during the single-legged squat. AB - BACKGROUND: Numerous factors have been identified as potentially increasing the risk of anterior cruciate ligament injury in the female athlete. However, differences between the sexes in lower extremity coordination, particularly hip control, are only minimally understood. HYPOTHESIS: There is no difference in kinematic or electromyographic data during the single-legged squat between men and women. STUDY DESIGN: Descriptive comparison study. METHODS: We kinematically and electromyographically analyzed the single-legged squat in 18 intercollegiate athletes (9 male, 9 female). Subjects performed five single-legged squats on their dominant leg, lowering themselves as far as possible and then returning to a standing position without losing balance. RESULTS: Women demonstrated significantly more ankle dorsiflexion, ankle pronation, hip adduction, hip flexion, hip external rotation, and less trunk lateral flexion than men. These factors were associated with a decreased ability of the women to maintain a varus knee position during the squat as compared with the men. Analysis of all eight tested muscles demonstrated that women had greater muscle activation compared with men. When each muscle was analyzed separately, the rectus femoris muscle activation was found to be statistically greater in women in both the area under the linear envelope and maximal activation data. CONCLUSIONS: Under a physiologic load in a position commonly assumed in sports, women tend to position their entire lower extremity and activate muscles in a manner that could increase strain on the anterior cruciate ligament. PMID- 12750143 TI - True aneurysm of the ulnar artery in a soccer goalkeeper: a case report and surgical considerations. PMID- 12750144 TI - Extensor carpi ulnaris tendon rupture in an ice hockey player: a case report. PMID- 12750145 TI - Isolated supraspinatus muscle paralysis after shoulder dislocation: a case report. PMID- 12750146 TI - Imaging of the elbow in the overhead throwing athlete. AB - Elbow injuries in athletes who perform overhead throwing motions often present diagnostic challenges because of the undue stresses and often chronic, repetitive patterns of injury. Accurate and efficient assessment of the injured elbow is essential to maximize functional recovery and expedite return to play. Radiographic evaluation should be tailored to the specific injury suspected and requires a thorough understanding of normal anatomic relationships as well as familiarity with common injuries affecting these athletes. PMID- 12750147 TI - Allograft transplantation in the knee: tissue regulation, procurement, processing, and sterilization. AB - Use of musculoskeletal allografts has become increasingly popular, with widespread use among knee surgeons. The advantages and disadvantages of their use have been documented. In the knee, allografts are used for ligament reconstruction, meniscal transplantation, and articular surface reconstruction. The purpose of this review is to present issues surrounding the allograft industry, including regulation of tissues and tissue banks and procurement, processing, sterilization, and storage of allograft tissue. Tissue bank regulation is ultimately under the jurisdiction and authority of the Food and Drug Administration; some individual states regulate tissue banks. The American Association of Tissue Banks is a scientific organization that encourages education, research, and voluntary accreditation of tissue banks. It promotes safety and standards for retrieval, processing, storage, and distribution of transplantable human tissue. Allograft tissues are generally harvested and processed aseptically, which may not prevent contamination. Tissue sterilization is difficult and controversial. Tissue banks historically have used one of two methods of sterilization, ethylene oxide or gamma radiation. Both methods have risks and benefits. Newer methods of sterilization are being developed. Allograft tissue that is not transplanted fresh can be freeze-dried or deep frozen for storage. Ultimately, allograft transplantation in the knee facilitates knee form and function and enhances the patient's quality of life. Orthopaedic surgeons who use allograft tissue must understand the tissue banking process to provide safe and effective tissues to their patients. PMID- 12750149 TI - Serum but not myocardial TNF-alpha concentration is increased in pacing-induced heart failure in rabbits. AB - In animals and patients with severe heart failure (HF), the serum tumor necrosis factor-alpha (TNF-alpha) concentration is increased. It is, however, still controversial whether or not such increased serum TNF-alpha originates from the heart itself or is of peripheral origin secondary to gastrointestinal congestion and increased endotoxin concentration. We therefore now examined TNF-alpha in serum, myocardium, and liver of sham-operated and HF rabbits. In nine rabbits in which HF was induced by left ventricular (LV) pacing at 400 beats/min for 3 wk, LV end-diastolic diameter was increased and systolic shortening fraction (9.4 +/- 1.0 vs. 28.5 +/- 1.3%, echocardiography, P < 0.05) was reduced. Serum TNF-alpha was higher in HF than in sham-operated rabbits (240 +/- 24 vs. 150 +/- 22 U/ml, WEHI-cell assay, P < 0.05). In the heart, TNF-alpha was located mainly in the vascular endothelium (immunohistochemistry), and TNF-alpha protein (920 +/- 160 vs. 900 +/- 95 U/g) did not differ between groups. In the liver of HF rabbits, hepatocytes expressed TNF-alpha, and TNF-alpha protein was increased compared with sham-operated rabbits (2,390 +/- 310 vs. 1,220 +/- 135 U/g, P < 0.05) and correlated to the number of hepatic leukocytes (r = 0.85) and serum TNF-alpha (r = 0.69). The intestinal endotoxin concentration was 24.5 +/- 1.2 vs. 17.0 +/- 3.1 endotoxin units/g wet wt (P < 0.05) in HF compared with sham-operated rabbits. In this HF model, serum but not myocardial TNF-alpha is increased. The increased serum TNF-alpha originates from peripheral sources. PMID- 12750148 TI - Urethral closure mechanisms under sneeze-induced stress condition in rats: a new animal model for evaluation of stress urinary incontinence. AB - The urethral closure mechanism under a stress condition induced by sneezing was investigated in urethane-anesthetized female rats. During sneezing, while the responses measured by microtip transducer catheters in the proximal and middle parts of the urethra increased, the response in the proximal urethra was almost negligible when the bladder response was subtracted from the urethral response or when the abdomen was opened. In contrast, the response in the middle urethra during sneezing was still observed after subtracting the bladder response or after opening the abdomen. These responses in the middle urethra during sneezing were significantly reduced approximately 80% by bilateral transection of the pudendal nerves and the nerves to the iliococcygeous and pubococcygeous muscles but not by transection of the visceral branches of the pelvic nerves and hypogastric nerves. The sneeze leak point pressure was also measured to investigate the role of active urethral closure mechanisms in maintaining total urethral resistance against sneeze-induced urinary incontinence. In sham-operated rats, no urinary leakage was observed during sneeze, which produced an increase of intravesical pressure up to 37 +/- 2.2 cmH2O. However, in nerve-transected rats urinary leakage was observed when the intravesical pressure during sneezing exceeded 16.3 +/- 2.1 cmH2O. These results indicate that during sneezing, pressure increases elicited by reflex contractions of external urethral sphincter and pelvic floor muscles occur in the middle portion of the urethra. These reflexes in addition to passive transmission of increased abdominal pressure significantly contribute to urinary continence mechanisms under a sneeze-induced stress condition. PMID- 12750150 TI - Developmental changes in spontaneous smooth muscle activity in the neonatal rat urinary bladder. AB - Changes in spontaneous activity of the urinary bladder during postnatal development were examined in muscle strips from the base and dome of bladders from 1- to 5-wk-old rats. Activity was analyzed using fast Fourier transformation (FFT), nonlinear cross prediction, and the Shannon entropy test. Spontaneous activity was not detected in strips from 1- to 5-day-old rats but was observed in 50% of strips from 6- to 7-day-old rats and was prominent in strips from 2-wk-old animals. FFT analysis revealed one peak in activity, which was significantly faster in the bladder base (0.21 +/- 0.03 Hz) than in the dome (0.08 +/- 0.01 Hz). A second peak at approximately 0.5 Hz was detected at 3-5 wk of age. Atropine but not tetrodotoxin decreased the amplitude of spontaneous contractions, whereas carbachol, a muscarinic agonist, unmasked or stimulated spontaneous activity. These data suggest that slow rhythmic activity observed previously in neonatal whole bladders is generated by pacemaker cells in the bladder base or dome. The emergence of faster activity in bladders from older animals may reflect the development of multiple pacemaker sites, which would reduce coordination within the bladder wall and improve storage function in the mature bladder. PMID- 12750151 TI - Attenuated defense response and low basal blood pressure in orexin knockout mice. AB - The perifornical area of the hypothalamus has been known as the center for the defense response, or "fight or flight" response, which is characterized by a concomitant rise in arterial blood pressure (AP), heart rate (HR), and respiratory frequency (Rf). We examined whether orexin, a recently identified hypothalamic neuropeptide, contributes to the defense response and basal cardiovascular regulation using orexin knockout mice. Microinjection of a GABA-A receptor antagonist, bicuculline methiodide (0.1-1 mM in 20 nl), to the perifornical area in urethane-anesthetized wild-type mice elicited dose-dependent increases in AP, HR, and Rf. Although similar changes were observed in orexin knockout mice, intensities were smaller and duration was shorter than those in wild-type mice. Moreover, in an awake and freely moving condition, telemeter indwelling orexin knockout mice showed diminished cardiovascular and behavioral responses to emotional stress in the resident-intruder test. We also found that basal AP in orexin knockout mice was significantly lower in both anesthetized (117 +/- 8 mmHg in wild type and 92 +/- 3 in knockout) and conscious (125 +/- 6 mmHg in wild type and 109 +/- 2 in knockout) conditions. alpha-Adrenergic blockade with prazosin or ganglion blockade with hexamethonium canceled the difference in basal AP. HR and cardiac contractile parameters by echocardiography did not differ between the two strains of mice. These results indicate lower sympathetic vasoconstrictor tone in knockout mice. The present study suggests that orexin-containing neurons in the perifornical area play a role as one of the efferent pathways of defense response and also operate as a regulator of AP at basal condition by activating sympathetic outflow. PMID- 12750152 TI - Positive correlation of skeletal muscle UCP3 mRNA levels with overweight in male, but not in female, rats. AB - The objective of this study was to investigate the sex-dependent regulation of skeletal muscle uncoupling protein (UCP)3 mRNA expression in response to overweight and its relationship with serum levels of free fatty acids, leptin, and insulin. Two obesity models were used: rats made obese by feeding them with a cafeteria diet for 14 wk, and postcafeteria overweight rats fed a chow diet for 10 wk after consuming the cafeteria diet for 14 wk. The effects of 24-h fasting were studied in postcafeteria rats and their age-matched controls. The cafeteria rats ate a high-fat diet and attained an excess body weight that was higher in females (+59%) than in males (+39%). A trend to higher induction of abdominal muscle UCP3 mRNA in male rats than in females after cafeteria diet was apparent (+116% increase vs. +26% increase). Postcafeteria male but not female rats still showed the tendency to have increased UCP3 mRNA levels relative to their age matched controls. A linear regression analysis showed a significant positive correlation of the UCP3 mRNA levels with overweight and with serum levels of leptin and insulin in males, but not in females, and no correlation with serum free fatty acid levels. A subsequent correlation analysis and a multiple linear regression analysis showed that overweight was the only parameter actually related to UCP3 mRNA levels in males. Fasting-induced upregulation of muscle UCP3 mRNA levels was higher in males (5- to 7-fold) than in females (3- to 4-fold). Our results point to the existence of sex-associated differences in the control of muscle UCP3 expression in response to overweight and fasting, with an impaired induction in female rats under both conditions. The correlation of abdominal muscle UCP3 mRNA expression with overweight in males could be related to their relative resistance to gain weight after chronic overeating of a cafeteria diet, by the purported role of UCP3 in the regulation of lipid utilization. PMID- 12750154 TI - Identification of differentially expressed genes representing dendritic cell precursors and their progeny. AB - The development of dendritic cells (DCs) from hematopoietic progenitors is not well understood. Using a spleen-derived long-term culture (LTC) system, it has been possible to continuously generate DCs from progenitors maintained in culture. The nonadherent LTC-DC population is composed of 2 major subsets. These are the small LTC-DC or DC precursors and their progeny, the large LTC-DCs that phenotypically resemble immature DCs. In this study, subtracted cDNA libraries were generated containing sequences differentially expressed in small or large LTC-DCs. Differential screening was then used on plated library clones to select genes expressed in either the small or the large cell population. Real-time polymerase chain reaction (PCR) has been used to verify the selection procedure for several genes of particular interest. Known genes isolated from subtracted libraries were related to stages in DC development and supported previous findings regarding the function of small and large LTC-DCs. Large LTC-DCs expressed a number of immunologically important genes encoding CD86, CCR1, osteopontin, and lysozyme. Small LTC-DCs resembled progenitor DCs expressing genes related to the organization of the cytoskeleton, the regulation of antigen processing, and a number of mitochondrial and ribosomal proteins. Novel transcripts were isolated from small and large LTC-DC-subtracted libraries that could encode novel proteins important in DC development. This study describes changes in gene expression related to the development of CD11c+CD11b+ major histocompatibility complex 2 low (MHC2lo) CD8alpha- DCs from precursors in a stroma-dependent culture system in the absence of exogenous cytokines. PMID- 12750153 TI - Imatinib improves but may not fully reverse the poor prognosis of patients with CML with derivative chromosome 9 deletions. AB - Deletions of the derivative chromosome 9 occur in a subset of patients with Philadelphia chromosome-positive chronic myeloid leukemia (CML) and are associated with a poor prognosis on standard drug therapy. However, it is currently unknown if the presence of deletions influences the response to imatinib, an Abl-specific tyrosine kinase inhibitor, that has recently shown excellent hematologic and cytogenetic responses in patients with CML. We, therefore, compared hematologic and cytogenetic responses with imatinib in 397 patients with CML, and survival and progression in 354 of these patients, according to deletion status and disease phase. We found no difference in survival between patients with and without deletions, contrasting with previous reports in cohorts with a lower proportion of patients treated with imatinib. However, the time to disease progression on imatinib treatment was significantly shorter for patients with deletions, both in chronic phase (P =.02) and advanced phases (P =.02). Moreover, both in chronic phase and more advanced phases of CML, hematologic and cytogenetic responses were uniformly lower in patients with deletions, with significant differences seen for hematologic response (P =.04), for major cytogenetic response (P =.008) in chronic phase, and for hematologic response in advanced phases (P =.007) of CML. This finding suggests that differences in survival may become apparent with longer follow-up. PMID- 12750155 TI - Peripheral T-cell lymphomas unspecified presenting in the skin: analysis of prognostic factors in a group of 82 patients. AB - In the present study the clinicopathologic and immunophenotypic features of 82 patients with a CD30- peripheral T-cell lymphoma, unspecified, presenting in the skin were evaluated. The purpose of this study was to find out whether subdivision of these lymphomas on the basis of cell size, phenotype, or presentation with only skin lesions is clinically relevant. The study group included 46 primary cutaneous CD30- large cell lymphomas and 17 small/medium sized T-cell lymphomas as well as 17 peripheral T-cell lymphomas with both skin and extracutaneous disease at the time of diagnosis. Patients with primary cutaneous small- or medium-sized T-cell lymphomas had a significantly better prognosis (5-year-overall survival, 45%) than patients with primary cutaneous CD30- large T-cell lymphomas (12%) and patients presenting with concurrent extracutaneous disease (12%). The favorable prognosis in this group with primary cutaneous small- or medium-sized T-cell lymphomas was particularly found in patients presenting with localized skin lesions expressing a CD3+CD4+CD8- phenotype. In the primary cutaneous T-cell lymphoma (CTCL) group and in the concurrent group, neither extent of skin lesions nor phenotype had any effect on survival. Our results indicate that peripheral T-cell lymphomas, unspecified, presenting in the skin have an unfavorable prognosis, irrespective of the presence or absence of extracutaneous disease at the time of diagnosis, cell size, and expression of a CD4+ or CD8+ phenotype. The only exception was a group of primary cutaneous small- or medium-sized pleomorphic CTCLs with a CD3+CD4+CD8- phenotype and presenting with localized skin lesions. PMID- 12750156 TI - Regulation of plasminogen activation: a role for melanotransferrin (p97) in cell migration. AB - We recently reported that human recombinant melanotransferrin (p97) presents a high transport rate across the blood-brain barrier that might involve the low density lipoprotein receptor-related protein (LRP). We now report new interactions between p97 and another LRP ligand, the urokinase plasminogen activator (uPA) complex. By using biospecific interaction analysis, both pro-uPA and plasminogen are shown to interact with immobilized p97. Moreover, the activation of plasminogen by pro-uPA is increased by soluble p97. Because the uPA system plays a crucial role in cell migration, both in cancer and in angiogenesis, we also measured the impact of both endogenous membrane-bound and exogenous p97 on cell migration. The monoclonal antibody L235 (which recognizes a conformational epitope on p97) inhibited the migration of human microvascular endothelial cells (HMECs-1) and of human melanoma SK-MEL-28 cells, indicating that endogenous membrane-bound p97 could be associated with this process. In addition, low concentrations of exogenous p97 (10 and 100 nM) inhibited HMEC-1 and SK-MEL28 cell migration by more than 50%. These results indicate that membrane-bound and soluble p97 affect the migration capacity of endothelial and melanoma cells and suggest that p97 could be involved in the regulation of plasminogen activation by interacting with pro-uPA and plasminogen. PMID- 12750157 TI - The "stomatin" gene and protein in overhydrated hereditary stomatocytosis. AB - In overhydrated hereditary stomatocytosis (OHSt), Coomassie- and silver-stained polyacrylamide gels show an apparently complete deficit of the 32-kDa membrane protein, stomatin. We have used an antistomatin antibody to examine peripheral blood films, bone marrow, splenic tissue, and hepatic tissue from these patients by immunocytochemistry. This technique revealed that, in fact, some red cells did show positive stomatin immunoreactivity; and consistent with this result, Western blot analysis of the red cell membranes confirmed that about one twentieth to one fiftieth of the normal amount of stomatin was in fact present. Flow cytometry, combining immunoreactive quantitation of stomatin expression with thiazole orange staining for reticulocytes, showed that in OHSt, it was the young cells that had more stomatin. Magnetic-activated cell separation studies, using beads to which an anti-transferrin receptor antibody was conjugated, confirmed that in OHSt there was a correspondence between expression of stomatin and the transferrin receptor. Immunocytochemistry and Western blotting revealed that in OHSt patients, the protein was present in spleen, liver, neutrophils, platelets, monocytes, and about 50% of the peripheral lymphocytes, with the same distribution as in healthy controls. Neither Southern blots, nor direct sequencing of multiple subclones of the cDNA, nor sequencing of amplicons from genomic DNA revealed any significant abnormality in stomatin gene sequence in these patients. The deficiency of stomatin from red cells appears to be due to a loss of stomatin from these red cells on maturation in the bone marrow and in the circulation. PMID- 12750158 TI - Signaling through JAM-1 and alphavbeta3 is required for the angiogenic action of bFGF: dissociation of the JAM-1 and alphavbeta3 complex. AB - Growth factor-induced neovascularization has received a great deal of attention because it is fundamental to the growth and metastasis of solid tumors. This multistep process requires extensive signaling through growth factor receptors and integrins. Among the integrins involved in this process, integrin alphavbeta3 is specific to basic fibroblast growth factor (bFGF)-induced angiogenesis. Here we show that junctional adhesion molecule 1/A (JAM-1/A) and alphavbeta3 form a complex in the absence of bFGF. JAM-1, which is normally localized at the cell cell junctions of quiescent endothelial cells, redistributes to the cell surface on bFGF treatment. Blockage of the extracellular domain of JAM-1 inhibits bFGF induced endothelial cell morphology, proliferation, and angiogenesis. Additionally, mutation in the JAM-1 cytoplasmic domain blocks bFGF-induced mitogen-activated protein (MAP) kinase activation and ablates its ability to induce endothelial cell tube formation, suggesting that signaling through JAM-1 is key to bFGF-induced signaling. Immunoprecipitation analysis suggests that bFGF signaling dissociates the JAM-1/ alphavbeta3 complex, allowing for signaling through JAM-1 and alphavbeta3. In addition, blockage of either JAM-1 or alphavbeta3 inhibits bFGF-induced MAP kinase activation. Thus, our results suggest that signaling through JAM-1 and alphavbeta3 is necessary for bFGF induced angiogenesis. PMID- 12750159 TI - ALK activation by the CLTC-ALK fusion is a recurrent event in large B-cell lymphoma. AB - We present 3 cases of large B-cell lymphoma (LBCL) with a granular cytoplasmic staining for anaplastic lymphoma kinase (ALK). All of the cases showed striking similarities in morphology and immunohistochemical profile characterized by a massive monomorphic proliferation of CD20-/CD138+ plasmablast-like cells. In one of the cases, initially diagnosed as a null-type anaplastic large cell lymphoma (ALCL), the B-cell phenotype became evident only at recurrence. Fluorescent in situ hybridization (FISH) and molecular studies led to the detection of a CLTC ALK rearrangement in all 3 cases, without any evidence of full-length ALK receptor expression. The associated t(2;17)(p23;q23) was demonstrated in the karyotype of 2 cases. Although a similar CLTC-ALK aberration was previously identified in ALK-positive T-/null cell ALCL and inflammatory myofibroblastic tumor, its association with ALK-positive LBCL seems to be specific and intriguing. PMID- 12750160 TI - Disease-associated dendritic cells respond to disease-specific antigens through the common heat shock protein receptor. AB - The most abundant intracellular proteins, heat shock proteins (HSPs), serve as molecular chaperones for regulatory and maturation pathways. Diverse families of HSPs have been shown to bind antigenic peptides and to play major roles in innate and adaptive immune responses through the common HSP receptor, CD91. HIV-1+ patients with Kaposi sarcoma (KS) were matched for CD4 count and HIV-1 RNA viral load to HIV-1+ patients without Kaposi sarcoma (and negative for Kaposisarcoma associated herpesvirus). We then investigated the pathways used by tumor lysates, viral lysates, and viral particles in their activation. In particular, we observed immune responses after HSP depletion using antitumor antibiotics and blockade of the common HSP receptor, CD91. Despite the impaired functional capacity of dendritic cells (DCs) derived from patients with KS, DCs retain the ability to prime the adaptive arm of the immune system through the common HSP receptor, leading to phenotypic activation and stimulation of tetramer-positive CD8+ cytotoxic T cells. We also show that interferon-producing plasmacytoid DCs are selectively depleted in KS-positive compared with matched KS-negative HIV-1 infected patients. Functionally impaired DCs can effectively cross-present immune responses through the common HSP receptor. These results have important implications for the etiopathogenesis of KS and for the development and design of any compounds, including vaccines, derived from cellular lysates. PMID- 12750161 TI - High-dose radioimmunotherapy versus conventional high-dose therapy and autologous hematopoietic stem cell transplantation for relapsed follicular non-Hodgkin lymphoma: a multivariable cohort analysis. AB - We performed a multivariable comparison of 125 consecutive patients with follicular lymphoma (FL) treated at our centers with either high-dose radioimmunotherapy (HD-RIT) using 131I-anti-CD20 (n = 27) or conventional high dose therapy (C-HDT) (n = 98) and autologous hematopoietic stem cell transplantation. The groups were similar, although more patients treated with HD RIT had an elevated pretransplantation level of lactate dehydrogenase (41% versus 20%, P =.03) and elevated international prognostic score (41% versus 19%, P =.02). Patients treated with HD-RIT received individualized therapeutic doses of 131I-tositumomab (median, 19.7 GBq [531 mCi]) to deliver 17 to 31 Gy (median, 27 Gy) to critical organs. Patients treated with C-HDT received total body irradiation plus chemotherapy (70%) or chemotherapy alone (30%). Patients treated with HD-RIT experienced improved overall survival (OS) (unadjusted hazard ratio [HR] for death = 0.4 [95% confidence interval (95% CI), 0.2-0.9], P =.02; adjusted HR, 0.3, P =.004) and progression-free survival (PFS) (unadjusted HR =.6 [95% C.I., 0.3-1.0], P =.06; adjusted HR, 0.5, P =.03) versus patients treated with C-HDT. The estimated 5-year OS and PFS were 67% and 48%, respectively, for HD-RIT and 53% and 29%, respectively, for C-HDT. One hundred-day treatment related mortality was 3.7% in the HD-RIT group and 11% in the C-HDT group. The probability of secondary myelodysplastic syndrome/acute myeloid leukemia (MDS/AML) was estimated to be.076 at 8 years in the HD-RIT group and.086 at 7 years in the C-HDT group. HD-RIT may improve outcomes versus C-HDT in patients with relapsed FL. PMID- 12750162 TI - Mechanisms of donor-specific transfusion tolerance: preemptive induction of clonal T-cell exhaustion via indirect presentation. AB - Induction of transplantation tolerance to alloantigens without general immunosuppression remains an enduring challenge. Injecting a donor-specific transfusion (DST) of spleen cells together with blocking alphaCD154 antibody prior to graft transplantation is an effective way to induce long-lived graft acceptance. Using a novel T-cell receptor (TCR) transgenic (Tg) model of CD4+ T cell-mediated rejection, this study sheds new insights into the cellular basis for enhanced graft survival induced by DST and alphaCD154. The study shows that DST and alphaCD154 induce an early, robust, abortive expansion of the Tg T cells that results in profound anergy. This is contrasted with the more delayed, regional, productive response elicited by an allogeneic graft. Studies show that the induction of tolerance to the allograft induced by DST is mediated by indirect presentation by host antigen-presenting cells. Based on these observations, we conclude that DST and alphaCD154 preemptively tolerize the alloreactive T-cell compartment to prohibit subsequent responses to the immunogenic allograft. PMID- 12750163 TI - The HIF family member EPAS1/HIF-2alpha is required for normal hematopoiesis in mice. AB - Hypoxic stress plays a role in pathophysiologic states such as myocardial infarction and cerebral vascular events as well as in normal physiologic conditions including development and hematopoiesis. Members of the hypoxia inducible factor (HIF) family function as transcriptional regulators of genes involved in the hypoxic response. After generating adult mice that globally lack endothelial PAS domain protein 1 (EPAS1, also known as HIF-2alpha/HRF/HLF/MOP3), the second member of the HIF family, characterization of the hematopoietic cell population indicated that the loss of EPAS1/HIF-2alpha resulted in pancytopenia. Using bone marrow reconstitution experiments of lethally irradiated hosts, we have defined the extent and site of hematopoietic impairment in the EPAS1/HIF 2alpha null mice. These data suggest a critical role for EPAS1/HIF-2alpha in maintaining a functional microenvironment in the bone marrow for effective hematopoiesis. PMID- 12750164 TI - Iron, manganese, and cobalt transport by Nramp1 (Slc11a1) and Nramp2 (Slc11a2) expressed at the plasma membrane. AB - Mutations in the Nramp1 gene (Slc11a1) cause susceptibility to infection by intracellular pathogens. The Nramp1 protein is expressed at the phagosomal membrane of macrophages and neutrophils and is a paralog of the Nramp2 (Slc11a2) iron transporter. The Nramp1 transport mechanism at the phagosomal membrane has remained controversial. An Nramp1 protein modified by insertion of a hemagglutinin epitope into the predicted TM7/8 loop was expressed at the plasma membrane of Chinese hamster ovary cells as demonstrated by immunofluorescence and surface biotinylation. Experiments in Nramp1HA transfectants using the metal sensitive fluorophors calcein and Fura2 showed that Nramp1HA can mediate Fe2+, Mn2+, and Co2+ uptake. Similar results were obtained in transport studies using radioisotopic 55Fe2+ and 54Mn2+. Nramp1HA transport was dependent on time, temperature, and acidic pH, occurring down the proton gradient. These experiments suggest that Nramp1HA may be a more efficient transporter of Mn2+ compared to Fe2+ and a more efficient Mn2+ transporter than Nramp2HA. The membrane topology and transport properties of Nramp1HA and Nramp2HA were indistinguishable, suggesting that Nramp1 divalent-metal transport at the phagosomal membrane is mechanistically similar to that of Nramp2 at the membrane of acidified endosomes. These results clarify the mechanism by which Nramp1 contributes to phagocyte defenses against infections. PMID- 12750165 TI - Ex vivo characterization of human CD8+ T subsets with distinct replicative history and partial effector functions. AB - After antigenic challenge, naive T lymphocytes enter a program of proliferation and differentiation during the course of which they acquire effector functions and may ultimately become memory cells. In humans, the pathways of effector and memory T-cell differentiation remain poorly defined. Here we describe the properties of 2 CD8+ T-lymphocyte subsets, RA+CCR7-27+28+ and RA+CCR7-27+28-, in human peripheral blood. These cells display phenotypic and functional features that are intermediate between naive and effector T cells. Like naive T lymphocytes, both subsets show relatively long telomeres. However, unlike the naive population, these T cells exhibit reduced levels of T-cell receptor excision circles (TRECs), indicating they have undergone additional rounds of in vivo cell division. Furthermore, we show that they also share effector-type properties. At equivalent in vivo replicative history, the 2 subsets express high levels of Fas/CD95 and CD11a, as well as increasing levels of effector mediators such as granzyme B, perforin, interferon gamma, and tumor necrosis factor alpha. Both display partial ex vivo cytolytic activity and can be found among cytomegalovirus-specific cytolytic T cells. Taken together, our data point to the presence of T cells with intermediate effector-like functions and suggest that these subsets consist of T lymphocytes that are evolving toward a more differentiated effector or effector-memory stage. PMID- 12750166 TI - Recombinant human activated protein C (rhAPC; drotrecogin alfa [activated]) has minimal effect on markers of coagulation, fibrinolysis, and inflammation in acute human endotoxemia. AB - Inflammatory and procoagulant host responses are closely related in sepsis. The protein C pathway serves as a regulatory pathway with anti-inflammatory and anticoagulant properties. Recently, recombinant human activated protein C (rhAPC) was shown to reduce mortality in severe sepsis. Nevertheless, the effects of rhAPC in humans are still ill defined. The infusion of low endotoxin doses into humans provides a standardized model to study inflammatory and hemostatic mechanisms. Thus, we investigated whether rhAPC acts as an anticoagulant or anti inflammatory drug in human endotoxemia. There were 24 volunteers randomized to receive either 24 microg/kg per hour rhAPC or placebo intravenously for 8 hours. Lipopolysaccharide (LPS, 2 ng/kg) was administered 2 hours after starting the infusions. rhAPC decreased basal tissue factor (TF)-mRNA expression, and thrombin formation and action. In contrast, rhAPC did not significantly blunt LPS-induced thrombin generation. Consistently, rhAPC did not reduce LPS-induced levels of TF mRNA or D-dimer and had no effect on fibrinolytic activity or inflammation. Finally, endogenous APC formation was enhanced during endotoxemia and appeared to be associated with inflammation rather than thrombin formation. In conclusion, even low-grade endotoxemia induces significant protein C activation. Infusion of rhAPC decreases "spontaneous" activation of coagulation but does not blunt LPS induced, TF-mediated coagulation in healthy volunteers, which is in contrast to a number of anticoagulants. PMID- 12750167 TI - BAALC expression predicts clinical outcome of de novo acute myeloid leukemia patients with normal cytogenetics: a Cancer and Leukemia Group B Study. AB - Cytogenetic aberrations are important prognostic factors in acute myeloid leukemia (AML). Of adults with de novo AML, 45% lack cytogenetic abnormalities, and identification of predictive molecular markers might improve therapy. We studied the prognostic impact of BAALC (Brain And Acute Leukemia, Cytoplasmic), a novel gene involved in leukemia, in 86 de novo AML patients with normal cytogenetics who were uniformly treated on Cancer and Leukemia Group B 9621. BAALC expression was determined by comparative real-time reverse transcriptase polymerase chain reaction in pretreatment blood samples, and patients were dichotomized at BAALC's median expression into low and high expressers. Low expressers had higher white counts (P =.03) and more frequent French-American British M5 morphology (P =.007). Compared to low expressers, high BAALC expressers showed significantly inferior overall survival (OS; median, 1.7 vs 5.8 years, P =.02), event-free survival (EFS; median, 0.8 vs 4.9 years, P =.03), and disease-free survival (DFS; median, 1.4 vs 7.3 years, P =.03). Multivariable analysis confirmed high BAALC expression as an independent risk factor. For high BAALC expressers the hazard ratio of an event for OS, EFS, and DFS was respectively 2.7, 2.6, and 2.2. We conclude that high BAALC expression predicts an adverse prognosis and may define an important risk factor in AML with normal cytogenetics. PMID- 12750168 TI - Mechanisms for T-cell selective cytotoxicity of arabinosylguanine. AB - Nelarabine, prodrug of arabinosylguanine (ara-G), has demonstrated T lymphoblastic antileukemic activity in cell lines and in the clinic. To investigate the mechanism for lineage-specific toxicity, the effects of ara-G were compared in CEM (T-lymphoblast), Raji (B-lymphoblast), and ML-1 (myeloid) cell lines. CEM cells were the most sensitive to ara-G-induced apoptosis and accumulated the highest levels of ara-G triphosphate (ara-GTP). However, compared with myeloid and B-lineage cell lines, CEM cells incorporated fewer ara-G molecules-which were at internucleotide positions in all 3 cell lines- into DNA. Ara-G induced an S-phase arrest in both Raji and ML-1, while in CEM the S-phase cells decreased with a concomitant increase in the sub-G1 population. Within 3 hours of ara-G treatment, the levels of soluble Fas ligand (sFasL) in the medium increased significantly in CEM cultures. In parallel, an induction of FasL gene expression was observed by real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Pretreatment of CEM cells with a Fas antagonistic antibody inhibited ara-G-mediated cell death. These results demonstrate that high ara-GTP accumulation in T cells results in an S phase-dependent apoptosis induced by ara G incorporation into DNA, which may lead to a T cell-specific signal for the induction and liberation of sFasL. Subsequently, the sFasL induces an apoptotic response in neighboring non-S-phase cells. In contrast, myeloid and B cells accumulated lower levels of ara-GTP and arrested in S phase, blocking any apoptotic signaling. PMID- 12750169 TI - Heme and lipid peroxides in hemoglobin-modified low-density lipoprotein mediate cell survival and adaptation to oxidative stress. AB - Low-density lipoprotein (LDL) oxidation mediated by a variety of catalysts in atherosclerotic lesions plays a crucial role in the genesis and evolution of atherosclerotic plaques. In this study we focused on oxidative properties of hemoglobin (Hb)-modified LDL because Hb is present in atherosclerotic lesions. Under low oxygen tensions Hb was previously found to modify apolipoprotein B100 with covalent binding of Hb fragments and formation of electronegative LDL particles (LDL-). Here we show that HbLDL is highly susceptible to oxidation, but is not cytotoxic to vascular cells, as was found for LDL- isolated from human plasma. HbLDL and LDL- have similar levels of oxidized lipid products and low uptake rates; however, the virtual absence of HbLDL-induced toxicity depends on a marked adaptive oxidative stress response. This was evidenced by a time- and dose dependent induction of heme oxygenase (HO-1). Cell survival was significantly decreased in the presence of HO-1 inhibitor, tin protoporphyrin (SnPPIX). HO-1 induction by HbLDL increased resistance of cells to toxic doses of hemin or t BuOOH. The high sensitivity to oxidation and HO-1 induction was largely dependent on lipid hydroperoxides and heme associated with HbLDL. Reduction of pre-existing lipid peroxides using ebselen delayed HbLDL kinetics and inhibited HO-1 induction. Moreover, heme inactivation or its degradation inhibited HO-1 induction and provided an additive inhibitory effect to ebselen. We conclude that Hb-catalyzed reactions may modulate vascular cell survival and oxidative stress adaptation due to the presence of peroxides and heme, thus providing a possible mechanism for the evolution of atherosclerotic and hemorrhagic lesions. PMID- 12750170 TI - Transgenic mice overexpressing erythropoietin adapt to excessive erythrocytosis by regulating blood viscosity. AB - Severe elevation of red blood cell number is often associated with hypertension and thromboembolism resulting in severe cardiovascular complications. However, some individuals such as high altitude dwellers cope well with an increased hematocrit level. We analyzed adaptive mechanisms to excessive erythrocytosis in our transgenic (tg) mice that, due to hypoxia-independent erythropoietin (Epo) overexpression, reached hematocrit values of 0.8 to 0.9 without alteration of blood pressure, heart rate, or cardiac output. Extramedullar erythropoiesis occurred in the tg spleen, leading to splenomegaly. Upon splenectomy, hematocrit values in tg mice decreased from 0.89 to 0.62. Tg mice showed doubled reticulocyte counts and an increased mean corpuscular volume. In tg mice, plasma volume was not elevated whereas blood volume was up to 25% of the body weight compared with 8% in wild-type (wt) siblings. Although plasma viscosity did not differ between tg and wt mice, tg whole-blood viscosity increased to a lower degree (4-fold) than expected from corresponding hemoconcentrated wt blood (8 fold). This moderate increase in viscosity is explicable by the up to 3-fold higher elongation of tg erythrocytes at physiologic shear rates. Apart from the nitric oxide-mediated vasodilation we reported earlier, adaptation to high hematocrit levels in tg mice involves regulated elevation of blood viscosity by increasing erythrocyte flexibility. PMID- 12750171 TI - Myeloid cell expansion elicited by the progression of spontaneous mammary carcinomas in c-erbB-2 transgenic BALB/c mice suppresses immune reactivity. AB - Transgenic female mice expressing the transforming rat oncogene c-erbB-2 (HER 2/neu) under the mouse mammary tumor virus (MMTV) promoter (BALB-neuT) spontaneously develop mammary carcinomas with a progression resembling that of human breast cancer. In these mice, activating antitumor immunotherapy fails to induce T cell-mediated cytotoxicity, suggesting a suppression of the immune response. We found a direct correlation between tumor multiplicity and an increased proportion of Gr-1+ (Ly6G)/Mac-1+(CD11b)/ER-MP12+(CD31) immature myeloid cells in the peripheral blood (PB) and spleen, suggesting that tumor load profoundly affects overall BALB-neuT hematopoiesis. In fact, myeloid colony formation was increased in bone marrow (BM) and spleen. The immature myeloid cells displayed suppressive activity on host T lymphocytes, which progressively failed to respond to alloantigens and CD3 triggering, while maintaining the ability to proliferate in response to nonspecific mitogens. Transplantation of normal BM into BALB-neuT mice readily resulted in hypertrophic hematopoiesis with myeloid cell expansion. This persistent influence of the tumor was mediated through the release of vascular endothelial growth factor (VEGF) but not granulocyte-macrophage colony-stimulating factor (GM-CSF), and was down-modulated when tumor load was reduced but not when BM was transplanted. Together, the data obtained in the BALB-neuT model of naturally occurring carcinogenesis show that tumor-associated immune suppression is secondary to a more general alteration of host hematopoiesis, conditioned by tumor-secreted soluble factors. PMID- 12750172 TI - TNF-alpha and IFN-gamma are overexpressed in the bone marrow of Fanconi anemia patients and TNF-alpha suppresses erythropoiesis in vitro. AB - In Fanconi anemia (FA) C mice tumor necrosis factor alpha (TNF-alpha) and interferon gamma (IFN-gamma) have key roles in the pathogenesis of bone marrow failure. In FA subjects TNF-alpha was found to be increased in the serum and overproduced by patient-derived B-cell lines. In acquired aplastic anemia, a disease in which, similarly to FA, marrow failure occurs, TNF-alpha and IFN-gamma act as late mediators of the stem cell damage and are overexpressed in patient marrow lymphocytes. This study evaluated in marrow mononuclear cells (MNCs) of patients with FA, the expression of negative modulators of the hematopoiesis, such as TNF-alpha, IFN-gamma, macrophage inflammatory protein 1alpha (MIP 1alpha), and surface Fas ligand, and the role of TNF-alpha on FA erythropoiesis in vitro. TNF-alpha and IFN-gamma were significantly overexpressed in stimulated marrow MNCs of FA patients as compared to healthy controls. MIP-1alpha and Fas ligand were undetectable in patients and controls. In bone marrow cultures, the addition of anti-TNF-alpha increased the size and significantly increased the number of erythroid colony-forming units and erythroid burst-forming units grown from FA patients but not from healthy controls. This indicates that FA subjects have a marrow TNF-alpha activity that inhibits erythropoiesis in vitro. TNF-alpha has a relevant role in the pathogenesis of erythroid failure in FA patients. PMID- 12750173 TI - CXCR3-mediated chemotaxis of human T cells is regulated by a Gi- and phospholipase C-dependent pathway and not via activation of MEK/p44/p42 MAPK nor Akt/PI-3 kinase. AB - The chemokines CXCL9, 10, and 11 exert their action via CXC chemokine receptor-3 (CXCR3), a receptor highly expressed on activated T cells. These interferon gamma (IFNgamma)-induced chemokines are thought to be crucial in directing activated T cells to sites of inflammation. As such, they play an important role in several chronic inflammatory diseases including ulcerative colitis, multiple sclerosis, artherosclerosis, and delayed-type hypersensitivity reactions of the skin. In this study, we first demonstrate that in COS-7 cells heterologously expressing CXCR3, CXCL11 is a potent activator of the pertussis toxin (PTX)-sensitive p44/p42 mitogen-activated protein kinase (MAPK) and Akt/phosphatidylinositol 3 kinase (PI3K) pathways. Next, we show that these signal transduction pathways are also operative and PTX sensitive in primary human T cells expressing CXCR3. Importantly, abrogation of these signaling cascades by specific inhibitors did not block the migration of T cells toward CXCR3 ligands, suggesting that MAPK and Akt activation is not crucial for CXCR3-mediated chemotaxis of T cells. Finally, we demonstrate that CXCR3-targeting chemokines control T-cell migration via PTX sensitive, phospholipase C pathways and phosphatidylinositol kinases other than class I PI3Kgamma. PMID- 12750174 TI - Inhibition of bcr-abl gene expression by small interfering RNA sensitizes for imatinib mesylate (STI571). AB - Bcr-Abl proteins are effective inducers of the leukemic phenotype in chronic myeloid leukemia (CML) and distinct variants of acute lymphoblastic leukemia (ALL). Targeting bcr-abl by treatment with the selective tyrosine kinase inhibitor imatinib has proved to be highly efficient for controlling leukemic growth. However, it is unclear whether imatinib is sufficient to eradicate the disease because of primary or secondary resistance of leukemic cells. Therefore, targeting Bcr-Abl with an alternative approach is of great interest. We demonstrate that RNA interference (RNAi) with a breakpoint-specific short interfering RNA (siRNA) is capable of decreasing Bcr-Abl protein expression and of antagonizing Bcr-Abl-induced biochemical activities. RNAi selectively inhibited Bcr-Abl-dependent cell growth. Furthermore, bcr-abl-homologous siRNA increased sensitivity to imatinib in Bcr-Abl-overexpressing cells and in a cell line expressing the imatinib-resistant Bcr-Abl kinase domain mutation His396Pro, thereby antagonizing 2 of the major mechanisms of resistance to imatinib. PMID- 12750175 TI - Expression and function of KIR and natural cytotoxicity receptors in NK-type lymphoproliferative diseases of granular lymphocytes. AB - Using monoclonal antibodies (mAbs) specific for different natural killer (NK) receptors, we studied the lymphocyte population from 18 patients with NK-type lymphoproliferative disease of granular lymphocytes (LDGL). The analysis of both resting and cultured NK cell populations demonstrated that these patients are frequently characterized by NK cells displaying a homogeneous staining with given anti-killer Ig-like receptor (anti-KIR) mAb (11 of 18 patients). In most patients NK cells were characterized by the CD94/NKG2A+ phenotype, whereas only a minor fraction of the cases expressed CD94/NKG2C. In 7 of these patients we could also assess the function of the various NK receptors. Remarkably those KIR molecules that, in each patient, homogeneously marked the NK cell expansion were found to display an activating function as determined by cross-linking with specific anti KIR mAb. The KIR genotype analysis performed in 13 of 18 cases revealed that in NK-type LDGL certain activating KIRs, as well as certain infrequent KIR genotypes, were detected with higher frequencies as compared to previously analyzed healthy donors. Moreover, most KIR genotypes included multiple genes coding for activating KIRs. The analysis of non-HLA-specific triggering receptors indicated that the natural cytotoxicity receptors (NKp46, NKp30) were expressed at significantly low levels in freshly drawn NK cells from most patients analyzed. However, in most instances the expression of NKp46 and NKp30 could be up-regulated on culture in interleukin 2. Our data indicate that in NK-LDGL the expanded subset is frequently characterized by the expression of a given activating KIR, suggesting a direct role for these molecules in the pathogenetic mechanisms of this disorder. PMID- 12750176 TI - High-penetrance mouse model of acute promyelocytic leukemia with very low levels of PML-RARalpha expression. AB - Transgenic mice expressing PML-RARalpha in early myeloid cells under control of human cathepsin G regulatory sequences all develop a myeloproliferative syndrome, but only 15% to 20% develop acute promyelocytic leukemia (APL) after a latent period of 6 to 14 months. However, this transgene is expressed at very low levels in the bone marrow cells of transgenic mice. Because the transgene includes only 6 kb of regulatory sequences from the human cathepsin G locus, we hypothesized that sequences required for high-level expression of the transgene might be located elsewhere in the cathepsin G locus and that a knock-in model might yield much higher expression levels and higher penetrance of disease. We, therefore, targeted a human PML-RARalpha cDNA to the 5' untranslated region of the murine cathepsin G gene, using homologous recombination in embryonic stem cells. This model produced a high-penetrance APL phenotype, with more than 90% of knock-in mice developing APL between 6 and 16 months of age. The latent period and phenotype of APL (including a low frequency of an interstitial deletion of chromosome 2) was similar to that of the previous transgenic model. Remarkably, however, the expression level of PML-RARalpha in bone marrow cells or APL cells was less than 3% of that measured in the low-penetrance transgenic model. Although the explanation for this result is not yet clear, one hypothesis suggests that very low levels of PML-RARalpha expression in early myeloid cells may be optimal for the development of APL in mice. PMID- 12750177 TI - Immunotherapy of tumors with vaccine based on quail homologous vascular endothelial growth factor receptor-2. AB - The breaking of immune tolerance of "self-antigens" associated with angiogenesis is an attractive approach to cancer therapy by active immunity. We used vascular endothelial growth factor receptor-2 (VEGFR-2) as a model antigen to explore the feasibility of the immunotherapy with a vaccine based on a xenogeneic homologous protein. To test this concept, we prepared a quail homologous VEGFR-2 protein vaccine (qVEGFR) based on quail VEGFR-2. At the same time, a protein vaccine based on the corresponding ligand-binding domain of mouse self-VEGFR-2 (mVEGFR) was also prepared and used as a control. We found that immunotherapy with qVEGFR was effective at protective and therapeutic antitumor immunity in several solid and hematopoietic tumor models in mice. Autoantibodies against mouse VEGFR-2 (Flk 1) were identified by Western blot analysis and enzyme-linked immunosorbent assay (ELISA). Anti-VEGFR antibody-producing B cells were detectable by ELISPOT. Endothelial deposition of immunoglobulins developed within tumor. VEGF-mediated endothelial cell proliferation was inhibited in vitro by immunoglobulins from qVEGFR-immunized mice. Antitumor activity was caused by the adoptive transfer of the purified immunoglobulins. Antitumor activity and production of autoantibodies against Flk-1 could be abrogated by the depletion of CD4+ T lymphocytes. Angiogenesis was apparently inhibited within the tumors, and the vascularization of alginate beads was also reduced. No marked toxicity was found in the immunized mice. The observations may provide a vaccine strategy for cancer therapy through the induction of autoimmunity against the growth factor receptor associated with angiogenesis in a cross-reaction with single xenogeneic homologous protein. PMID- 12750179 TI - Rapid recovery from T lymphopenia by CD28 superagonist therapy. AB - Slow recovery of T-cell numbers and function contributes to the high incidence of life-threatening infections after cytotoxic cancer therapies. We have tested the therapeutic potential of a novel class of superagonistic CD28-specific antibodies that induce polyclonal T-cell proliferation without T-cell receptor engagement in an experimental rat model of T lymphopenia. We show that in lethally irradiated, bone marrow-reconstituted hosts, CD28 superagonist is able to dramatically accelerate repopulation by a small inoculum of mature, allotype-marked T cells. CD28-driven recovery of CD4 cells was superior to that of CD8 T cells. CD28 superagonist- expanded CD4 T cells had maintained repertoire diversity and were functional both in vitro and in vivo, suggesting that treatment with a human CD28 specific superagonist will protect T-lymphopenic patients from opportunistic infections. PMID- 12750178 TI - Treatment of canine cyclic neutropenia by lentivirus-mediated G-CSF delivery. AB - Cyclic neutropenia is a rare disease that occurs both in humans and gray collie dogs and is characterized by recurrent severe neutropenia leading to bacterial infections and shortened life expectancy. Daily injections of recombinant granulocyte colony-stimulating factor (rG-CSF) are effective in shortening the period of severe neutropenia and reducing infections. After demonstrating that rG CSF induced elevated neutrophil production in an affected dog, cytokine administration was stopped and 109 infectious units (IUs) of a lentivirus pseudotyped with vesicular stomatitis virus G protein (VSV-G) encoding canine G CSF cDNA was administered intramuscularly. Serial blood cell counts showed elevated neutrophil production for longer than 17 months. Although neutrophil counts continued to cycle, the range at nadirs was from 3710 to 5300 cells/microL, well above the nadirs before lentivirus administration. After the injection of lentivirus, mean neutrophil counts +/- SD were 12 460 +/- 4240 cells/microL, significantly increased over both pretreatment values of 3040 +/- 2540 cells/microL(P <.0001) and neutrophil counts during G-CSF administration of 10 290 +/- 4860 cells/microL(P <.007). The changes in blood counts from lentivirus injection were associated with absence of clinical signs of infection and fever. The gray collie continued to gain weight and was no longer housed in a pathogen-free environment. Genomic DNA from muscle at injection sites was positive for provirus, whereas gonad, lung, spleen, heart, liver, kidney, leukocytes, and noninjected muscle samples were all negative for provirus. Thus, intramuscular administration of lentivirus encoding G-CSF provided sustained therapeutic levels of neutrophils, suggesting this approach may be applied for long-term treatment of patients with cyclic and other neutropenias. PMID- 12750180 TI - Finding the age of the patient's heart. PMID- 12750181 TI - Chronic obstructive pulmonary disease. PMID- 12750182 TI - Effect of passive smoking on health. PMID- 12750183 TI - The therapeutic effects of meditation. PMID- 12750184 TI - Public response to the Isaacs report will slow neurological research. PMID- 12750185 TI - Inquiry finds brains were removed without consent. PMID- 12750186 TI - Improvements in the NHS are patchy, report says. PMID- 12750187 TI - Heart surgeons are to be rated according to bypass surgery success. PMID- 12750188 TI - UK agency unveils plans to cut infusion device errors. PMID- 12750190 TI - Inertia on folic acid has caused thousands of unnecessary deaths. PMID- 12750194 TI - United States and Germany are keen to see tobacco agreement watered down. PMID- 12750196 TI - China is still not open enough about SARS, says WHO. PMID- 12750203 TI - UK government supports measures to cut drug prices in developing countries. PMID- 12750204 TI - SARS in China spreads from Beijing to poorer inland provinces. PMID- 12750205 TI - Environmental tobacco smoke and tobacco related mortality in a prospective study of Californians, 1960-98. AB - OBJECTIVE: To measure the relation between environmental tobacco smoke, as estimated by smoking in spouses, and long term mortality from tobacco related disease. DESIGN: Prospective cohort study covering 39 years. SETTING: Adult population of California, United States. PARTICIPANTS: 118 094 adults enrolled in late 1959 in the American Cancer Society cancer prevention study (CPS I), who were followed until 1998. Particular focus is on the 35 561 never smokers who had a spouse in the study with known smoking habits. MAIN OUTCOME MEASURES: Relative risks and 95% confidence intervals for deaths from coronary heart disease, lung cancer, and chronic obstructive pulmonary disease related to smoking in spouses and active cigarette smoking. RESULTS: For participants followed from 1960 until 1998 the age adjusted relative risk (95% confidence interval) for never smokers married to ever smokers compared with never smokers married to never smokers was 0.94 (0.85 to 1.05) for coronary heart disease, 0.75 (0.42 to 1.35) for lung cancer, and 1.27 (0.78 to 2.08) for chronic obstructive pulmonary disease among 9619 men, and 1.01 (0.94 to 1.08), 0.99 (0.72 to 1.37), and 1.13 (0.80 to 1.58), respectively, among 25 942 women. No significant associations were found for current or former exposure to environmental tobacco smoke before or after adjusting for seven confounders and before or after excluding participants with pre-existing disease. No significant associations were found during the shorter follow up periods of 1960-5, 1966-72, 1973-85, and 1973-98. CONCLUSIONS: The results do not support a causal relation between environmental tobacco smoke and tobacco related mortality, although they do not rule out a small effect. The association between exposure to environmental tobacco smoke and coronary heart disease and lung cancer may be considerably weaker than generally believed. PMID- 12750206 TI - Prevalence of hardcore smoking in England, and associated attitudes and beliefs: cross sectional study. AB - OBJECTIVE: To quantify the prevalence and characteristics of hardcore smokers in England. DESIGN: Cross sectional survey. SETTING: Interview in respondents' household. PARTICIPANTS: 7766 adult cigarette smokers. MAIN OUTCOME MEASURES: Hardcore smoking defined by four criteria (less than a day without cigarettes in the past five years; no attempt to quit in the past year; no desire to quit; no intention to quit), all of which had to be satisfied. RESULTS: Some 16% of all smokers were categorised as hardcore. Hardcore smoking was associated with nicotine dependence, socioeconomic deprivation, and age, rising from 5% in young adults aged 16-24 to 30% in those aged >or= 65 years. Hardcore smokers displayed distinctive attitudes towards and beliefs about smoking. In particular they were likely to deny that smoking affected their health or would do so in the future. Prevalence of hardcore smoking was almost four times higher than in California. CONCLUSION: Hardcore smoking presents a serious challenge to public health efforts to reduce the prevalence of smoking, but the proportion of hardcore smokers does not necessarily increase as overall prevalence in a population declines. More hardcore smokers could be persuaded to quit, but this will require interventions that are targeted to the particular needs and perceptions of both socially disadvantaged and older smokers. PMID- 12750207 TI - Mortality in young offenders: retrospective cohort study. AB - OBJECTIVES: To estimate overall and cause specific standardised mortality ratios in young offenders. DESIGN: Comparison of mortality data in cohort of young offenders. SETTINGS: State of Victoria, Australia. SUBJECTS: Cohort of young offenders aged 10-20 years with a first custodial sentence from 1 January 1988 to 31 December 1999. MAIN OUTCOME MEASURES: Deaths ascertained by matching with the national death index, a database containing records of all deaths in Australia since 1980. Death rates in the reference Victorian population used to calculate standardised mortality ratios. RESULTS: The offender cohort comprised 2621 men and 228 women with 11 333 person years of observation. The median age of first detention was 17.9 years for men and 18.4 years for women. Median follow up was 3.3 years for men and 1.4 years for women. Overall standardised mortality ratio adjusted for age (expressed as a ratio) was 9.4 (95% confidence interval 7.4 to 11.9) for men and 41.3 (20.2 to 84.7) for women. Cause specific standardised mortality ratios for men were 25.7 (17.9 to 36.9) for drug related causes, 9.2 (5.8 to 15) for suicide, and 5.7 (3.6 to 9.2) for non-intentional injury. A quarter of drug related deaths in men aged 15-19 years were in offenders. CONCLUSIONS: Social policies for young offenders should address both the prevalent drug and mental health problems as well the high levels of social disadvantage. PMID- 12750208 TI - New strategies for increasing the detection of HIV: analysis of routine data. PMID- 12750209 TI - Perinatal characteristics and risk of rheumatoid arthritis. PMID- 12750210 TI - Systematic review of scope and quality of electronic patient record data in primary care. AB - OBJECTIVE: To systematically review measures of data quality in electronic patient records (EPRs) in primary care. DESIGN: Systematic review of English language publications, 1980-2001. DATA SOURCES: Bibliographic searches of medical databases, specialist medical informatics databases, conference proceedings, and institutional contacts. STUDY SELECTION: Studies selected according to a predefined framework for categorising review papers. DATA EXTRACTION: Reference standards and measurements used to judge quality. RESULTS: Bibliographic searches identified 4589 publications. After primary exclusions 174 articles were classified, 52 of which met the inclusion criteria for review. Selected studies were primarily descriptive surveys. Variability in methods prevented meta analysis of results. Forty eight publications were concerned with diagnostic data, 37 studies measured data quality, and 15 scoped EPR quality. Reliability of data was assessed with rate comparison. Measures of sensitivity were highly dependent on the element of EPR data being investigated, while the positive predictive value was consistently high, indicating good validity. Prescribing data were generally of better quality than diagnostic or lifestyle data. CONCLUSION: The lack of standardised methods for assessment of quality of data in electronic patient records makes it difficult to compare results between studies. Studies should present data quality measures with clear numerators, denominators, and confidence intervals. Ambiguous terms such as "accuracy" should be avoided unless precisely defined. PMID- 12750212 TI - Variable presentation of Brugada syndrome: lessons from three generations with syncope. PMID- 12750213 TI - ABC of interventional cardiology: percutaneous coronary intervention. I: history and development. PMID- 12750211 TI - New pacing technologies for heart failure. PMID- 12750214 TI - Effect of regression to the mean on decision making in health care. PMID- 12750215 TI - Case fatality rates after hospital admission for stroke. Covariates were not measured. PMID- 12750216 TI - Case fatality rates after hospital admission for stroke. Definition of stroke defines stroke mortality. PMID- 12750217 TI - Open letter to health ministers at Framework Convention on Tobacco Control. PMID- 12750218 TI - Thyroid function tests and hypothyroidism. Restoring serum TSH to reference range should be goal of replacement. PMID- 12750219 TI - Thyroid function tests and hypothyroidism. Reducing concentrations further would be harmful. PMID- 12750220 TI - Computerised evidence based guidelines in primary care. Interactive decision support is here and essential for 21st century primary care. PMID- 12750221 TI - Computerised evidence based guidelines in primary care. Computerised decision support and reflection in action. PMID- 12750228 TI - Neurolinguistic programming: the keys to success. PMID- 12750223 TI - Findings for looked after children are not generalisable. PMID- 12750229 TI - A career in forensic child and adolescent psychiatry. PMID- 12750230 TI - Tackling harassment. PMID- 12750231 TI - Paediatric clinical pharmacology training should not be left in its infancy. PMID- 12750233 TI - Counterpoint: From animal models to prevention of colon cancer. Criteria for proceeding from preclinical studies and choice of models for prevention studies. AB - Corpet and Pierre (D. E. Corpet and F. Pierre, Cancer Epidemiol. Biomark. Prev., 12: 391-400, 2003) have reviewed the prevention studies made with the azoxymethane rat and Min mouse colon cancer models, and have shown that many agents reduce the numbers of these experimental tumors. They suggest that agents with preventive activity with little or no toxicity should be evaluated in clinical intervention studies without delay. I think that the decision to proceed to a clinical trial is more complex, and involves an understanding of the safety of the agent and of the strength and consistency of the preclinical data. However, I am also impressed by the wide range of agents that have been found to affect the development of colon cancer in animals. This suggests that human colon cancer may be the consequence of many different dietary and lifestyle deficiencies, a view supported by the observation that normal mice develop colon cancer when fed diets deficient in several food components known to prevent tumors with the azoxymethane rat model [Newmark, H. L. et al., Carcinogenesis (Lond.), 22: 1871-1875, 2001]. There is a clear need to evaluate the preventive effects of additional combinations of these agents, identified perhaps from the Corpet and Pierre review (D. E. Corpet and F. Pierre, Cancer Epidemiol. Biomark. Prev., 12: 391-400, 2003), or from actual human high-risk diets. With diets that increase the risk of "spontaneous cancer" in hand, the stage would be set for assessing the most effective ways to reduce colon cancer risk, again first with animal studies, then clinical trials, and then perhaps population studies. PMID- 12750232 TI - Point: From animal models to prevention of colon cancer. Systematic review of chemoprevention in min mice and choice of the model system. AB - The Apc(Min/+) mouse model and the azoxymethane (AOM) rat model are the main animal models used to study the effect of dietary agents on colorectal cancer. We reviewed recently the potency of chemopreventive agents in the AOM rat model (D. E. Corpet and S. Tache, Nutr. Cancer, 43: 1-21, 2002). Here we add the results of a systematic review of the effect of dietary and chemopreventive agents on the tumor yield in Min mice. The review is based on the results of 179 studies from 71 articles and is displayed also on the internet http://corpet.net/min.(2) We compared the efficacy of agents in the Min mouse model and the AOM rat model, and found that they were correlated (r = 0.66; P < 0.001), although some agents that afford strong protection in the AOM rat and the Min mouse small bowel increase the tumor yield in the large bowel of mutant mice. The agents included piroxicam, sulindac, celecoxib, difluoromethylornithine, and polyethylene glycol. The reason for this discrepancy is not known. We also compare the results of rodent studies with those of clinical intervention studies of polyp recurrence. We found that the effect of most of the agents tested was consistent across the animal and clinical models. Our point is thus: rodent models can provide guidance in the selection of prevention approaches to human colon cancer, in particular they suggest that polyethylene glycol, hesperidin, protease inhibitor, sphingomyelin, physical exercise, epidermal growth factor receptor kinase inhibitor, (+) catechin, resveratrol, fish oil, curcumin, caffeate, and thiosulfonate are likely important preventive agents. PMID- 12750234 TI - Breast cancer, birth cohorts, and Epstein-Barr virus: methodological issues in exploring the "hygiene hypothesis" in relation to breast cancer, Hodgkin's disease, and stomach cancer. AB - To address methodological issues in exploring a variant of the "hygiene hypothesis" that posits delayed infection by Epstein-Barr virus contributes to rising rates of breast cancer and Hodgkin's disease, we examined birth cohort trends in the incidence of both cancers plus stomach cancer, building on previously reported year-of-diagnosis cross-sectional associations of age standardized rates. Using published data from the United States Connecticut state cancer registry (1935-1998) for women for each cancer site, we obtained age specific incidence rates by birth cohort (1870-1874 to 1970-1974), along with age standardized incidence rates for selected calendar years (1935-1939, 1940-1944,., 1990-1994, 1995-1998). Clear secular trends in incidence rates, in the opposite direction, were evident for: (a) breast cancer and for Hodgkin's disease in young adults (increasing), and (b) stomach cancer (decreasing). Correlations between the incidence of breast cancer among women ages 50-54 and Hodgkin's disease among young adults (ages 20-24) were stronger for birth cohort (Pearson correlation, 0.85) than for cross-sectional analyses (Pearson correlation, 0.68). Stronger associations between the incidence of breast cancer and non-Hodgkin's disease were evident for birth cohort compared with cross-sectional analyses, findings consonant with (but not "proof" of) the hygiene hypothesis. One methodological implication is that tests of the hygiene hypothesis must take into account birth cohort effects and age at incidence of the outcomes under study; age-standardized cross-sectional analyses may be misleading. PMID- 12750235 TI - Association of markers of insulin and glucose control with subsequent colorectal cancer risk. AB - We evaluated the association of plasma insulin and other markers of insulin and glucose control with subsequent colorectal cancer. Incident colon (n = 132) and rectal (n = 41) cancer cases and matched controls (n = 346) were identified between baseline in 1989 and 2000 among participants in a community-based cohort in Washington County, Maryland. Circulating markers of insulin and glucose control were measured in baseline blood samples. Body mass index (BMI) and use of medications to treat diabetes mellitus were self-reported at baseline. Conditional logistic regression was used to estimate matched odds ratios (ORs). Compared with the lowest fourth, participants with insulin concentrations in the highest fourth were not at an increased risk of colorectal cancer [OR, 0.78; 95% confidence interval (CI), 0.45-1.35; P(trend) = 0.24]. Similarly, no associations were observed for the ratio of total cholesterol:HDL-cholesterol, triglycerides, and insulin-like growth factor binding protein 1. However, those in the highest fourth of glycosylated hemoglobin (HbA(1c)) level had a slightly increased risk of colorectal cancer (OR, 1.57; 95% CI, 0.94-2.60; P(trend) = 0.02). The OR of colorectal cancer was 1.70 (95% CI, 1.01-2.86; P(trend) = 0.08) comparing BMI >/=30 kg/m(2) to <25 kg/m(2). The OR of colorectal cancer was 2.43 (95% CI, 1.10 5.38) for the use of medications to treat diabetes. The associations of higher HbA(1c), higher BMI, and the use of medications to treat diabetes, with colorectal cancer lend support to the hypothesis that perturbations in insulin and glucose control may influence colorectal carcinogenesis. It is possible that HbA(1c), BMI, and the use of medications to treat diabetes, as a surrogate for protracted or severe type 2 diabetes mellitus, may have been better time-averaged indicators of hyperinsulinemia and hyperglycemia than the plasma markers that were measured once prediagnostically in a nonfasting population. PMID- 12750236 TI - Alcohol consumption, alcohol dehydrogenase 3 polymorphism, and colorectal adenomas. AB - Alcohol is a probable risk factor with regard to colorectal neoplasm and is metabolized to the carcinogen acetaldehyde by the genetically polymorphic alcohol dehydrogenase 3 (ADH3) enzyme. We evaluated whether the association between alcohol and colorectal adenomas is modified by ADH3 polymorphism. We recruited 433 cases with adenomatous polyps and 436 polyp-free controls among Caucasians undergoing endoscopy between 1995 and 2000. Frequency and amount of habitual alcohol consumption were assessed by beverage type, using a validated self administered food frequency questionnaire. All participants provided blood for genotyping of ADH3. Multivariate analyses adjusting for gender, age, and indication for endoscopy showed that alcohol increased the risk of colorectal adenomas among women [odds ratio (OR), 1.8; 95% confidence interval (CI), 1.0 3.2, >/=10 versus <1 drink/week]. Among men, the risk of adenomas was increased only for those consuming > 21 drinks/week (OR, 1.8; 95% CI, 0.9-3.8, compared with men drinking < 1 drink/week). Among subjects in the highest tertile of alcohol consumption, those with the ADH3*1/*1 genotype were at higher risk (OR, 1.8; 95% CI, 1.0-3.1) than those with other ADH3 genotypes (OR, 1.2; 95% CI, 0.7 1.9) when compared with those in the lowest tertile with ADH3*1/*2 or ADH3*2/*2 genotypes. In conclusion, our findings are consistent with results of other studies, suggesting that alcohol consumption elevates the risk of adenomatous colorectal polyps. ADH3 polymorphism may modify the association between alcohol consumption and colorectal adenomas. PMID- 12750237 TI - Decisional consideration of hereditary colon cancer genetic test results among Hong Kong chinese adults. AB - This study investigated the relationship between psychosocial factors and the decisional consideration of genetic testing of hereditary colon cancer. Attitudes and beliefs about genetic testing, anxiety and depression levels, coping style, and optimism were used as psychosocial independent variables. Sixty-two registrants (61% males and 39% females) of the Hereditary Gastrointestinal Cancer Registry of the Queen Mary Hospital in Hong Kong completed a mail survey. Mean age of the respondents was 42 years (SD = 9.92 years, range: 18-68 years). Correlational analyses and regression analyses were used to examine the relationships between the dependent and independent variables. Participants were concerned about the well-being and reactions of their significant others even more than their own well-being in their decisional consideration processes. Those who had higher perceived risks of being a mutated carrier and higher depression levels tended to emphasize more on the negative consequences of learning the test results and sharing them with relatives. Besides, those who believed that having cancer was attributable to personal (e.g., stress) rather than environmental factors considered that the negative consequences were relatively more important than the positive gains in sharing their results with relatives. Our participants tended to be relational or interdependent oriented in their decisional consideration processes related to genetic testing of colon cancer. This result is consistent with the established interdependent orientation of Chinese. Participants with higher risk perception focused more on the negative consequences of genetic testing. Psychological counseling might help these patients to cope with their concerns about being diagnosed as gene carriers after genetic testing. PMID- 12750238 TI - Parity, reproductive factors, and the risk of pancreatic cancer in women. AB - Incidence rates for pancreatic cancer are consistently lower in women than in men. Previous studies suggest that reproductive factors, particularly parity, may reduce pancreatic cancer risk in women. We examined parity, breast feeding history, age at first birth, menstrual factors, and exogenous hormone use in relation to pancreatic cancer risk in a prospective cohort study of women. Information on parity and other reproductive factors was assessed by questionnaires in 1976 and updated biennially. Multivariate relative risks were adjusted for cigarette smoking, body mass index, diabetes, and height. During 22 years of follow-up (1976-1998), 115,474 women contributed 2.4 million years of person time, and 243 cases of pancreatic cancer were identified. Compared with nulliparous women, the relative risk of pancreatic cancer was 0.86 [95% confidence interval (CI), 0.55-1.36] for women with 1-2 births, 0.75 (95% CI, 0.48-1.17) for 3-4 births, and 0.58 (95% CI, 0.34-0.98) for those with >/=5 births after adjusting for other factors. An analysis for linear trend indicates a 10% reduction in risk for each birth (P(trend) = 0.008). Other reproductive factors and exogenous hormone use were not significantly related to pancreatic cancer risk. In this large prospective cohort of women, parity was associated significantly with a reduced risk of pancreatic cancer. Additional studies should examine the physiological or hormonal changes underlying pregnancy or childbirth that may explain this finding. PMID- 12750239 TI - The XRCC1 399Gln polymorphism and the frequency of p53 mutations in Taiwanese oral squamous cell carcinomas. AB - DNA repair gene polymorphisms have been implicated as susceptibility factors in cancer development. It is possible that DNA repair polymorphisms may also influence the risk of gene mutation. The 399Gln polymorphism in the DNA repair gene XRCC1 has been indicated to have a contributive role in DNA adduct formation, sister chromatid exchange, and an increased risk of cancer development. Two hundred thirty-seven male oral squamous cell carcinomas (OSCCs) were included in a study to investigate the role of the XRCC1 194Trp, 280His, and 399Gln polymorphisms on p53 gene mutation. PCR-single-strand conformation polymorphism and DNA sequencing were used to analyze the conserved regions of the p53 gene (exons 5-9). The XRCC1 genotype was determined by PCR-RFLP. Nineteen (8.02%) of the 237 OSCCs had a Gln/Gln genotype. One hundred six (43.88%) of the 237 OSCCs showed p53 gene mutations at exons 5-9. The OSCC patients with a Gln/Gln genotype exhibited a significantly higher frequency of p53 mutation than those with an Arg/Gln and an Arg/Arg genotype. After adjustment for age, cigarette smoking, areca quid chewing, and alcohol drinking, the Gln/Gln genotype still showed an independent association with the frequency of p53 mutation (odd ratio, 4.50; 95% confidence interval, 1.52-13.36). The findings support the hypothesis that XRCC1 Arg399Gln amino acid change may alter the phenotype of the XRCC1 protein, resulting in a DNA repair deficiency. This study also suggests an important role for the XRCC1 399Gln polymorphism in p53 gene mutation in Taiwanese OSCCs. PMID- 12750240 TI - Antibodies against 5-hydroxymethyl-2'-deoxyuridine are associated with lifestyle factors and GSTM1 genotype: a report from the Malmo Diet and Cancer cohort. AB - Plasma autoantibodies (aAbs) against the oxidized DNA base derivative 5 hydroxymethyl-2'-deoxyuridine (5-HMdU) are potential biomarkers of cancer risk and oxidative stress. We examined their association with a number of cancer risk factors: smoking, alcohol habits, body fatness, and absence of the glutathione S transferases M1 and T1 (GSTM1 and GSTT1) in a sample from the population-based Malmo Diet and Cancer cohort (Sweden). This was a cross-sectional study of 264 men and 280 women, 46-67 years of age. Anti-5-HMdU aAb concentration was determined by an ELISA. Data on tobacco exposure were collected through a questionnaire. Alcohol consumption was estimated by a modified diet history method. Body fatness was assessed by a bioimpedance method. The absence or presence of genes coding for GSTM1 and GSTT1 was determined in granulocyte DNA by a multiplex PCR technique. aAb titers were significantly greater in those with high alcohol consumption. Current smokers lacking GSTM1, particularly men, had greater aAb titers compared with nonsmokers or persons expressing GSTM1. Body fatness was inversely associated with antibody titers in men. GSTT1 genotype was not associated with aAb titers. Overall, women had higher aAb titers than men. Adjustment for potential confounders (history of chronic diseases, anti inflammatory medication, and season of blood sampling) did not change the results. Our study shows that a high alcohol consumption, smoking in combination with lack of GSTM1, and low body fatness (in men) is associated with high titers of anti-5-HMdU aAbs in this population. PMID- 12750241 TI - Correlation of serum hormone concentrations in maternal and umbilical cord samples. AB - Evidence suggests that adult cancer risk of hormonally related tumors may be influenced by the in utero environment, and most speculation on the biological mechanism has focused on the hormonal component. Epidemiological studies investigating the biological nature of pregnancy and maternal factors associated with offspring's cancer risk have relied on maternal hormone measurements. The degree to which maternal hormone levels represent the fetal environment, however, is not widely known. Pregnancy estrogen, androstenedione, testosterone, dehydroepiandrosterone (DHEA), and DHEA-sulfate concentrations were measured in maternal and mixed umbilical cord sera from 86 singleton pregnancies. Spearman correlations between maternal and cord hormone levels generally ranged between 0.2 and 0.3. The correlation was 0.26 for estriol, the estrogen of highest concentration in pregnancy, and 0.27 for estradiol, the most biologically active estrogen. The correlations between mother and offspring for the estrogens and DHEA appeared similar for males and females, whereas there was a suggestion that the maternal-umbilical cord correlations for other androgens varied in magnitude by fetal sex, and all correlations appeared higher in pregnancies lasting <38 weeks compared with longer gestational lengths, although these stratified findings may have been attributable to chance. These data show a moderate degree of correlation in hormone concentrations between the maternal and fetal circulation. Studies using maternal hormone concentrations as a proxy for the fetal environment should consider the misclassification resulting with the use of this marker. PMID- 12750242 TI - Breast cancer risk reduction associated with the RAD51 polymorphism among carriers of the BRCA1 5382insC mutation in Poland. AB - The observed heterogeneity of breast cancer risk among women who carry the same BRCA1 mutation suggests the existence of modifying environmental and genetic factors. The product of the RAD51 gene functions with BRCA1 and BRCA2 in the repair of double-stranded DNA breaks. To establish whether polymorphic variation of RAD51 modifies risk for hereditary breast cancer, we conducted a matched case control study on 83 pairs of female carriers of the BRCA1 5382insC mutation. Cases consisted of women with breast cancer, and controls were women with the same mutation but who were unaffected. The frequency of the RAD51 135C variant allele was established in cases and controls using RFLP-PCR. The RAD51 135C allele was detected in 37% of unaffected and in 17% of affected BRCA1 carriers. Among 27 discordant matched pairs, the RAD51 135C allele was found in the healthy carrier on 22 occasions and in the affected carrier on only five occasions (odds ratio = 0.23; 95% confidence interval, 0.07-0.62; P = 0.0015). This finding suggests that RAD51 is a genetic modifier of breast cancer risk in BRCA1 carriers in the Polish population. It will be of interest to confirm this in other populations as well. PMID- 12750244 TI - Prediagnostic serum selenium concentration and the risk of recurrent colorectal adenoma: a nested case-control study. AB - Several studies have suggested that selenium may help to prevent colorectal neoplasia. To investigate the relation between prediagnostic serum selenium concentrations and colorectal adenomas, we conducted a nested case-control study using data from a large, multicenter, adenoma prevention trial. Cases comprised a total of 276 patients who developed a colorectal adenoma between the year 1 and year 4 follow-up exam. Controls were 276 patients who did not develop an adenoma during this time interval, matched to case subjects on age, sex, and clinical center. Total and bound selenium concentrations were measured from baseline or year 1 serum samples using instrumental neutron activation analysis. We estimated the odds ratios of colorectal adenoma in relation to serum selenium concentrations adjusting for age, clinical center, and sex. Compared with the lowest quintile, the odds ratio for the highest quintile was 0.76 (95% confidence interval, 0.44-1.30) for total selenium and 0.60 (95% confidence interval, 0.34 1.05) for bound selenium, and there was no apparent trend in risk (P for trend = 0.50 for total selenium and P for trend = 0.20 for bound selenium). Thus, our findings do not indicate a clear association between serum selenium concentrations and adenoma recurrence. PMID- 12750243 TI - Serum antibodies to JC virus, BK virus, simian virus 40, and the risk of incident adult astrocytic brain tumors. AB - Genomic sequences of the human polyomaviruses, JC virus (JCV) and BK virus (BKV), and simian virus 40 (SV40) have been reported from several types of human brain tumors, but there have been no population-based seroepidemiologic studies to evaluate the association between polyomavirus infection and brain tumors. We conducted a case-control study, nested within a prospective cohort, to investigate the association between antibodies to JCV, BKV, and SV40, as measured in serum collected 1-22 years before diagnosis and incident primary malignant brain tumors. Brain tumor cases (n = 44) and age-, gender-, and race-matched controls (n = 88) were identified from participants of two specimen banks in Washington County, Maryland. IgG antibodies to the capsid proteins of JCV and BKV were assessed using ELISAs. SV40-neutralizing antibodies were measured using plaque neutralization assays. Similar to the general population, the prevalence of JCV and BKV infection was high in our study population (77 and 85%, respectively). Antibodies to SV40 were less prevalent (11%). The odds ratio for subsequent brain tumor development was 1.46 [95% confidence interval (CI), 0.61 3.5] for JCV, 0.66 for BKV (95% CI, 0.22-1.95), and 1.00 for SV40 (95% CI, 0.30 3.32). Given the high prevalence of JCV and BKV infections and the millions who were potentially exposed to SV40 through contaminated polio vaccines, future studies should attempt to replicate these findings. PMID- 12750245 TI - Individual differences in nicotine intake per cigarette. AB - The increase in levels of blood nicotine that occurs from smoking a single cigarette, sometimes referred to as a "nicotine boost," is an individualized measure of how much nicotine has been extracted from smoking a cigarette. This study investigated the demographic, smoking status, and psychological predictors of nicotine boost in a sample of 190 treatment-seeking smokers. Boost was assessed by comparing plasma nicotine levels before and after participants smoked one of their own brand cigarettes ad libitum. Positive affect (mood) was a significant positive predictor of nicotine boost, controlling for baseline cotinine levels and cigarette brand (Federal Trade Commission) nicotine delivery. However the proportion of variability accounted for in the model was relatively small (5%). Future research on individual differences in nicotine boost is warranted to clarify the role of psychological, physiological, and cigarette related determinants. PMID- 12750246 TI - No association between green tea and the risk of gastric cancer: pooled analysis of two prospective studies in Japan. PMID- 12750247 TI - Plasma leptin is not associated with prostate cancer risk. PMID- 12750248 TI - Cyclooxygenase-2 expression is related to nuclear grade in ductal carcinoma in situ and is increased in its normal adjacent epithelium. AB - Cyclooxygenase-2 (COX-2) is emerging as an important cancer biomarker and is now an experimental target for solid tumor treatment.However, no study has exclusively focused on COX-2 expression in early lesions such as ductal carcinoma in situ (DCIS). We examined COX-2 expression by immunohistochemistry in 46 cases of women undergoing surgical resection for DCIS. We found that COX-2 expression was detected in 85% of all DCIS specimens, with increased COX-2 staining correlating with higher nuclear grade. Strikingly, COX-2 staining intensity in the normal adjacent epithelium was stronger than in the DCIS lesion itself. Our observations demonstrate that COX-2 is up-regulated in the normal adjacent epithelium and supports the hypothesis that the surrounding epithelial tissue is part of the disease process in DCIS. PMID- 12750249 TI - DeltaNp63alpha and TAp63alpha regulate transcription of genes with distinct biological functions in cancer and development. AB - The p63 gene shows remarkable structural similarity to the p53 and p73 genes. Because of two promoters, the p63 gene generates two types of protein isoforms, TAp63 and DeltaNp63. Each type yields three isotypes (alpha, beta, gamma) because of differential splicing of the p63 COOH terminus. The purpose of this study was to determine whether there is a functional link between the distinct p63 isotypes in their transcriptional regulation of downstream targets and their role in various cellular functions. TAp63alpha and DeltaNp63alpha adenovirus expression vectors were introduced into Saos2 cells for 4 and 24 h, and then gene profiling was performed using a DNA microarray chip analysis. Seventy-four genes (>2-fold change in expression) were identified that overlapped between two independent studies. Thirty-five genes were selected for direct expression testing of which 27 were confirmed by reverse transcription-PCR or Northern blot analysis. A survey of these genes shows that p63 can regulate a wide range of downstream gene targets with various cellular functions, including cell cycle control, stress, and signal transduction. Our study thus revealed p63 transcriptional regulation of many genes in cancer and development while often demonstrating opposing regulatory functions for TAp63alpha and DeltaNp63alpha. PMID- 12750250 TI - Haem, not protein or inorganic iron, is responsible for endogenous intestinal N nitrosation arising from red meat. AB - Many N-nitroso compounds (NOC) are carcinogens. In this controlled study of 21 healthy male volunteers, levels of NOC on a high (420 grams) red meat diet were significantly greater (P = 0.001) than on a low (60 grams) meat diet but not significantly greater when an equivalent amount of vegetable protein was fed. An 8-mg supplement of haem iron also increased fecal NOC (P = 0.006) compared with the low meat diet, but 35-mg ferrous iron had no effect. Endogenous N nitrosation, arising from ingestion of haem but not inorganic iron or protein, may account for the increased risk associated with red meat consumption in colorectal cancer. PMID- 12750251 TI - Adenoma multiplicity in irradiated Apc(Min) mice is modified by chromosome 16 segments from BALB/c. AB - Ionizing radiation (IR) is a well-characterized carcinogen in humans and mice. The BALB/c mouse strain is unusually sensitive to IR-induced tissue damage and cancer development in a range of organs, suggestive of a partial defect in DNA damage response. This has been confirmed by finding BALB/c-specific functional polymorphism in Prkdc, a gene on mouse chromosome 16 that encodes the catalytic subunit of DNA-dependent protein kinase. Prkdc(BALB) has been associated with increased susceptibility to IR-induced mammary and lymphatic neoplasia. Here, we provide evidence that chromosome 16 segments from BALB/c interact with Apc(Min) (multiple intestinal neoplasia) and specifically enhance IR-induced adenoma development in the upper part of the small intestine. PMID- 12750252 TI - BALB/c alleles for Prkdc and Cdkn2a interact to modify tumor susceptibility in Trp53+/- mice. AB - In mice heterozygous for p53 (Trp53(+/-)), the incidence of mammary tumors varies among strains, with C57BL/6 being resistant and BALB/c being susceptible. Mammary tumor phenotypes were examined in female Trp53(+/-) F1 mice (C57BL/6 x BALB/c;n = 19) and N2 backcross mice [(C57BL/6 x BALB/c) x BALB/c] (n = 224). Susceptibility to mammary tumors segregated as a dominant phenotype in F1 females, but a higher frequency and shorter latency in N2 mice indicated a contribution from recessive acting modifiers. Segregation of the hypomorphic BALB/c alleles for DNA-dependent protein kinase catalytic subunit (Prkdc) and p16(INK4A) (Cdkn2a) was analyzed in the N2 mice. The time to first tumor (considering all tumor types) was significantly different among the four genotype combinations (P = 0.01). Cdkn2a had a strong effect (P = 0.008) but was restricted to Prkdc(B/B) mice (P = 0.001), indicating a strong interaction between the loci. Differences in mammary tumor occurrence among genotypes for Prkdc and Cdkn2a in N2 mice were not statistically significant. This study indicates that BALB/c Prkdc and Cdkn2a alleles do modify tumor incidence in Trp53(+/-) mice and highlights the complexity of gene interaction effects in determining cancer phenotypes but discounts these alleles as major recessive loci contributing to spontaneous mammary tumor susceptibility. PMID- 12750253 TI - Adenoviral gene transfer of tumor necrosis factor-related apoptosis-inducing ligand overcomes an impaired response of hepatoma cells but causes severe apoptosis in primary human hepatocytes. AB - Ligands of the tumor necrosis factor family play key roles in liver pathogenesis. The ligand tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is unique, because it is thought to be nontoxic to normal cells while killing a broad range of tumor cells. However, hepatocellular carcinoma is considered resistant to soluble TRAIL treatment. Therefore, a direct gene transfer of TRAIL to malignant cells is part of an alternative delivery strategy. We show that an adenoviral gene transfer (Ad-TRAIL) overcomes an impaired response of hepatocellular carcinoma cell lines to soluble TRAIL, but the transduction of primary human hepatocytes revealed a high number of apoptotic cells. Our data imply that Ad-TRAIL administration in vivo must either be restricted to tumor tissue or controlled by a tumor-specific promoter to avoid severe liver damage in human trials. PMID- 12750254 TI - p29ING4 and p28ING5 bind to p53 and p300, and enhance p53 activity. AB - We identified and characterized two new ING family genes, p29ING4 and p28ING5,coding for two proteins of 249 and 240 amino acids, respectively. Both p29ING4 and p28ING5 proteins have a plant homeodomain finger motif also found in other ING proteins, and which is common in proteins involved in chromatin remodeling. p29ING4 or p28ING5 overexpression resulted in a diminished colony forming efficiency, a decreased cell population in S phase, and the induction of apoptosis in a p53-dependent manner. Both p29ING4 and p28ING5 activate the p21/waf1 promoter, and induce p21/WAF1 expression. p29ING4 and p28ING5 enhance p53 acetylation at Lys-382 residues, and physically interact with p300, a member of histone acetyl transferase complexes, and p53 in vivo. These results indicate that p29ING4 and p28ING5 may be significant modulators of p53 function. PMID- 12750256 TI - Identification of mucin-depleted foci in the unsectioned colon of azoxymethane treated rats: correlation with carcinogenesis. AB - We tested the association between aberrant crypt foci (ACF) and tumor induction by feeding azoxymethane-induced rats (15 mg/kg x 2, s.c.) with synbiotics (Raftilose Synergy 1, a derivative of inulin, 10% of the diet, along with lactobacilli and bifidobacteria). After 16 weeks of feeding, synbiotics significantly increased ACF multiplicity. On the contrary, after 32 weeks, synbiotics significantly decreased intestinal tumors. When the same unsectioned colon used for ACF determination was stained with high-iron diamine Alcian blue, foci of crypts with scarce or absent mucins were identified. We defined these lesions as mucin-depleted foci (MDF), and they were visible in all azoxymethane treated rats and correlated with tumor induction (MDF/colon: 8.2 +/- 0.9 and 3.8 +/- 0.9 in controls and synbiotic-fed rats, respectively, P < 0.01; crypts/MDF: 12.2 +/- 2 and 6.4 +/- 1 in controls and synbiotic-fed rats, respectively, P < 0.05, means +/- SE, n = 7). There were fewer MDF/colon than ACF, and they were histologically more dysplastic than mucinous lesions identified as ACF in high iron diamine Alcian blue-stained colon. In conclusion, MDF may be premalignant lesions that predict colon carcinogenesis. PMID- 12750255 TI - Neurotensin stimulates protein kinase C-dependent mitogenic signaling in human pancreatic carcinoma cell line PANC-1. AB - Neuropeptides and their corresponding G protein-coupled receptors are increasingly implicated in the autocrine/paracrine stimulation of growth of human cancers. Using K-Ras mutated human pancreatic ductal adenocarcinoma cell line PANC-1 as a model system, we demonstrate that neurotensin (NT) induced translocation of phosphorylated extracellular signal-regulated kinases (ERK-1 and ERK-2) to the nucleus, rapid dose-dependent activation of dual-specificity mitogen and ERK-1 and ERK-2 kinase-1/2 (MEK-1/2), and striking stimulation of c Raf-1 but not pan-Ras. Furthermore, treatment of PANC-1 cells with protein kinase C (PKC) inhibitors, GF-1 and Ro 31-8220, completely abrogated NT-induced ERK-1 and ERK-2 activation, and significantly attenuated NT-induced c-Raf-1 stimulation. Interestingly, NT did not stimulate epidermal growth factor receptor transactivation, and epidermal growth factor receptor tyrosine kinase or Src inhibitors did not affect NT-induced ERK activation in PANC-1 cells. Our results indicate that NT potently stimulates c-Raf-1-MEK-ERK in PANC-1 cells through a PKC-dependent signaling pathway. Furthermore, we show that NT-induced DNA synthesis in PANC-1 cells is ERK-dependent. Finally, we demonstrate that NT stimulated clonal growth of PANC-1 cells in semisolid medium, which is abrogated by both GF-1 and the MEK-1/2 inhibitor, U0126. Collectively our results suggest that PKC-mediated stimulation of ERK-1 and ERK-2 play a pivotal role in NT induced growth of PANC-1 cells harboring activating K-Ras mutation. PMID- 12750257 TI - Enhancing major histocompatibility complex class I antigen presentation by targeting antigen to centrosomes. AB - Several strategies that increase proteasomal degradation of antigen have been shown to improve MHC class I presentation of antigen. Because recent studies have demonstrated that the centrosome is a subcellular compartment rich in proteasomes, we hypothesized that targeting a tumor antigen to centrosomal compartments would enhance both the MHC class I presentation of antigen and the vaccine potency. We, therefore, created a chimera of gamma-tubulin, an established centrosomal marker, with a model tumor antigen, human papillomavirus type 16 (HPV-16) E7, in a DNA vaccine. The linkage of gamma-tubulin to E7 targeted antigen to centrosomal compartments, resulted in enhanced MHC class I presentation of E7, and led to a marked increase in the number of E7-specific CD8(+) T-cell precursors as well as a potent CD4-independent antitumor effect against an E7-expressing tumor cell line, TC-1. In addition, vaccination with gamma-tubulin/E7 DNA in transporter associated with antigen presentation (TAP)-1 knockout mice revealed that the enhancement of E7-specific CD8(+) T-cell immune responses is TAP-1-dependent. Our data suggest that the centrosome may be an important locus for MHC class I antigen processing and that targeting antigen to the centrosome can improve DNA vaccine potency. PMID- 12750258 TI - Perillyl alcohol as a chemopreventive agent in N-nitrosomethylbenzylamine-induced rat esophageal tumorigenesis. AB - Perillyl alcohol (POH) is a monoterpene found in lavender, spearmint, and cherries. Phase I clinical trials with this agent have shown a favorable toxicity profile and preliminary data indicate some chemotherapeutic efficacy in advanced cancers. Animal studies have demonstrated the ability of POH to inhibit tumorigenesis in the mammary gland, liver, and pancreas. Although the precise mechanism of action is unclear, POH has been shown to inhibit the farnesylation of small G-proteins, including Ras, up-regulate the mannose-6-phosphate receptor, and induce apoptosis. Previous studies in our laboratory using the rat model of squamous cell carcinoma of the esophagus have shown that a specific Ha-ras codon 12 mutation is important for tumor promotion and progression. Given the limited toxicity of POH in humans, its proven efficacy in several animal models and its potential to inhibit Ha-ras farnesylation, we conducted an animal study to evaluate the efficacy of POH as a chemopreventive agent for squamous cell carcinoma of the esophagus. Male Fischer-344 rats were treated s.c. with 0.25 mg/kg b.w. of N-nitrosomethylbenzylamine three times a week for 5 weeks. Three days after the final carcinogen dose, they were started either on control diet or diets containing 0.5 or 1.0% POH. At 25 weeks, the animals were sacrificed, and esophageal tumors were counted. Animals fed either dose of POH showed a significant increase in dysplasia when compared with controls (P < 0.05) and a nonsignificant trend toward increased tumor multiplicity. Additionally, 1.0% POH did not affect Ras membrane localization. These data indicate that POH has a weakly promoting effect early in nitrosamine-induced esophageal tumorigenesis and suggest that POH may not be an effective chemopreventive agent for esophageal cancer in humans. PMID- 12750259 TI - Disruption of protein kinase Ceta results in impairment of wound healing and enhancement of tumor formation in mouse skin carcinogenesis. AB - We have generated a mouse strain lacking protein kinase C (PKC) eta to evaluate its significance in epithelial organization and tumor formation. The PKCeta deficient mice exhibited increased susceptibility to tumor formation in two-stage skin carcinogenesis by single application of 7,12-dimethylbenz(a)anthracene (DMBA) for tumor initiation and repeated applications of 12-O tetradecanoylphorbol-13-acetate (TPA) for tumor promotion. The tumor formation was not enhanced by DMBA or TPA treatment alone, suggesting that PKCeta suppresses tumor promotion. Epidermal hyperplasia induced by topical TPA treatment was prolonged in the mutant mice. The enhanced tumor formation may be closely associated with the prolonged hyperplasia induced by topical TPA treatment. In the mutant mice, after inflicting injury by punch biopsy, wound healing on the dorsal skin, particularly reepithelialization, was significantly delayed and impaired in structure. Impairment of epithelial regeneration in wound healing indicates a possibility that PKCeta plays a role in maintenance of epithelial architecture. Homeostasis in epithelial tissues mediated by PKCeta is important for tumor formation in vivo. We propose that PKCeta is involved in tumor formation modulated by regulation of proliferation and remodeling of epithelial cells in vivo. PMID- 12750260 TI - Metabolic activation of temozolomide measured in vivo using positron emission tomography. AB - The purpose of this research was to quantitate and confirm the mechanism of in vivo metabolic activation of temozolomide. The secondary aims were to evaluate the tumor, normal tissue, and plasma pharmacokinetics of temozolomide in vivo, and to determine whether such pharmacokinetics resulted in tumor targeting. [(11)C]temozolomide kinetics were studied in men using positron emission tomography (PET). It has been postulated that temozolomide undergoes decarboxylation and ring opening in the 3-4 position to produce the highly reactive methyldiazonium ion that alkylates DNA. To investigate this, a dual radiolabeling strategy, with [(11)C]temozolomide separately radiolabelled in the 3-N-methyl and 4-carbonyl positions, was used. We hypothesized that (11)C in the C-4 position of [4-(11)C-carbonyl]temozolomide would be converted to [(11)C]CO(2) if the postulated mechanism of metabolic conversion was true resulting in lower [(11)C]temozolomide tumor exposure. Paired studies were performed with both forms of [(11)C]temozolomide in 6 patients with gliomas. Another PET scan with (11)C radiolabelled bicarbonate was performed and used to account for the metabolites of temozolomide using a data-led analytical approach. Plasma was analyzed for [(11)C]temozolomide and [(11)C]metabolites throughout the scan duration. Exhaled air was also sampled throughout the scan for [(11)C]CO(2). The percentage ring opening of temozolomide over 90 min was also calculated to evaluate whether there was a differential in metabolic breakdown among plasma, normal tissue, and tumor. There was rapid systemic clearance of both radiolabelled forms of [(11)C]temozolomide over 90 min (0.2 liter/min/m(2)), with [(11)C]CO(2) being the primary elimination product. Plasma [(11)C]CO(2) was present in all of the studies with [4-(11)C-carbonyl]temozolomide and in half the studies with [3-N (11)C-methyl]temozolomide. The mean contributions to total plasma activity by [(11)C]CO(2) at 10 and 90 min were 12% and 28% with [4-(11)C carbonyl]temozolomide, and 1% and 4% with [3-N-(11)C-methyl]temozolomide, respectively. There was a 5-fold increase in exhaled [(11)C]CO(2) sampled with [4 (11)C-carbonyl]temozolomide compared with [3-N-(11)C-methyl]temozolomide (P < 0.05). A decrease in tissue exposure [area under the curve between 0 and 90 min (AUC(0-90 min))] to [(11)C]temozolomide was also observed with [4-(11)C-carbonyl] temozolomide compared with [3-N-(11)C-methyl]temozolomide. Of potential therapeutic advantage was the higher [(11)C]radiotracer and [(11)C]temozolomide exposure (AUC(0-90 min)) in tumors compared with normal tissue. [(11)C]temozolomide ring opening over 90 min was less in plasma (20.9%; P < 0.05) compared with tumor (26.8%), gray matter (29.7%), and white matter (30.1%), with no differences (P > 0.05) between tumor and normal tissues. The significantly higher amounts of [(11)C]CO(2) sampled in plasma and exhaled air, in addition to the lower normal tissue and tumor [(11)C]temozolomide AUC(0-90 min) observed with [4-(11)C-carbonyl]temozolomide, confirmed the postulated mechanism of metabolic activation of temozolomide. A higher tumor [(11)C]temozolomide AUC(0-90 min) in tumors compared with normal tissue and the tissue-directed metabolic activation of temozolomide may confer potential therapeutic advantage in the activity of this agent. This is the first report of a clinical PET study used to quantify and confirm the in vivo mechanism of metabolic activation of a drug. PMID- 12750261 TI - Loss of coordinated androgen regulation in nonmalignant ovarian epithelial cells with BRCA1/2 mutations and ovarian cancer cells. AB - Epidemiological studies have implicated androgens in the etiology/progression of epithelial ovarian cancer. Because normal and malignant ovarian epithelial cells are growth inhibited by transforming growth factor (TGF) beta, we tested the ability of 5alpha-dihydrotestosterone (DHT) to modulate this response and the expression of TGF-beta receptor types I and II. Cells derived from the ovarian surface epithelium of women undergoing oophorectomy (n = 7) for nonovarian indications or with a germ-line BRCA1 or 2 mutation (n = 9), and from the ascitic fluid of patients with primary ovarian cancer (n = 8) were cultured with and without DHT. Cell proliferation after TGF-beta1 or vehicle treatment was determined, and transcripts for TGF-beta receptors were measured by quantitative reverse transcription-PCR. As low levels of androgen receptor were observed in the cultures, we also measured transcript levels for steroid receptor coactivators SRC-1, ARA70, and AIB1. TGF-beta1 inhibited growth in 12 of 13 cultures tested, and DHT generally reversed this effect, demonstrating that androgens can block TGF-beta-induced growth inhibition in both malignant and nonmalignant ovarian epithelial cells. Transcripts for TGF-beta receptors, SRC-1, and ARA70 were found to be coordinately regulated by androgen in control cells, but not in either malignant or BRCA1/2-positive cell cultures. These findings raise the possibility that by modulating steroid receptor coactivator expression, androgen might affect other hormonal responses and contribute to the initiation of ovarian cancer. PMID- 12750262 TI - Hormonal and dietary modulation of mammary carcinogenesis in mouse mammary tumor virus-c-erbB-2 transgenic mice. AB - Exogenous and dietary estrogens have been associated with modification of breast cancer risk. Mammary cancer model systems can be used to explore interactions between specific transgenes, and hormonal and dietary factors. Transgenic mice bearing the rat wild-type erbB-2 gene were used to study the effects of short term hormonal exposure [17beta-estradiol (E2) or tamoxifen] or a soy meal diet on mammary carcinogenesis. In mice fed a casein diet, mammary tumors developed at an earlier age after short-term E2 exposure during the early reproductive period. The median mammary tumor latency was shortest (29 weeks) for the high-dose estrogen as compared with the lowest dose of E2 treated or placebo control mice (33 and 37 weeks, respectively). The timing of short-term E2 exposure was also important, with the most significant changes observed in mice exposed to E2 between 8 and 18 weeks of age. E2 exposure was associated with the subsequent development of more aggressive tumors as determined by histologic grade, multifocal tumor development, stromal invasion, and pulmonary metastasis. In contrast, short-term tamoxifen-exposed mice generally failed to develop mammary tumors by 60 weeks of age. Mice fed a soy meal diet developed mammary tumors at a later age than casein-fed animals treated with E2 or placebo, whereas no differences were observed by diet for the tamoxifen-treated mice. Mammary tumor prevention was >80% in tamoxifen-treated mice on either diet. Novel histologic tumor types were identified, suggesting greater phenotypic diversity than described previously. Benign mammary gland morphogenesis was also significantly altered by short-term hormonal exposure or dietary factors, consistent with the modification of mammary tumor risk in specific treatment groups. Estrogenic modulation of the mammary tumor phenotype in wild-type erbB-2 transgenic mice was observed. Histologic tumor types and clinical aggressivity not reported previously in this transgenic model were noted, suggesting greater biologic heterogeneity than reported previously. In addition, dietary phytoestrogens modified mammary development and tumor latency, suggesting a need for greater stringency in dietary assignment for transgenic mouse models of mammary neoplasia. PMID- 12750263 TI - Estrogen receptor beta protein in human breast cancer: correlation with clinical tumor parameters. AB - The recent discovery of a second estrogen receptor (ER), designated ERbeta, raises pressing questions about its role in estrogen regulation of human breast cancer cells and its significance for the prediction of recurrence and treatment responses in clinical breast cancer. Most of what we know about ERbeta expression comes from studies examining a limited number of samples at the RNA level. We have now generated a monoclonal antibody useful for the detection of ERbeta at the protein level in archival, formalin-fixed breast tumors and have examined its expression using immunohistochemistry in a pilot series of 242 breast cancer patients. Coexpression of ERbeta and ERalpha was found in the majority of the tumors, with 76% of the tumors expressing ERbeta as determined by immunohistochemistry. ERalpha, but not ERbeta, was strongly associated with progesterone receptor expression, suggesting that ERalpha is the predominant regulator of this estrogen-induced gene in breast tumors. Although ERalpha expression was positively correlated with low tumor grade, diploidy, and low S phase fraction, all biological parameters of a good prognostic profile, ERbeta trended toward an association only with aneuploidy; no association with tumor grade or S-phase fraction was seen for ERbeta. We found that ERbeta expression does cause false positive readings for ERalpha. These results suggest that ERbeta expression is not a surrogate for ERalpha in clinical breast tumors and, as such, could be a useful biomarker in its own right. PMID- 12750264 TI - Breast cancer risk associated with genotypic polymorphism of the nonhomologous end-joining genes: a multigenic study on cancer susceptibility. AB - The role of the familial breast cancer susceptibility genes, BRCA1 and BRCA2, in the homologous recombination pathway for DNA double-strand break (DSB) repair suggests that the mechanisms involved in DNA DSB repair are of particular etiological importance during breast tumorigenesis. However, there is currently no evidence for an association between breast cancer and the other DSB repair pathway, the nonhomologous end-joining (NHEJ) pathway. It is possible that, because this DNA repair pathway is so crucial for mammalian cells to maintain genomic stability, any severe defects in it would result in serious outcomes, such as genomic instability and cell death, and block subsequent cell outgrowth and tumor formation. Thus, only subtle defects arising from low-penetrance alleles would escape lethality accumulating essential genetic changes and be associated with cancer formation, and the tumorigenic contribution of these alleles would become more obvious if individual putative high-risk genotypes of each NHEJ gene act jointly. Furthermore, this joint effect might be modified by specific environmental factors, and we hypothesized that estrogen exposure might be one such factor because estrogen is suggested to cause DNA DSBs, triggering breast tumorigenesis. Because single nucleotide polymorphisms (SNPs) are the most subtle genetic variation in the genome, to examine these hypotheses, we have genotyped 30 SNPs in all five NHEJ genes (Ku70, Ku80, DNA-PKcs, Ligase IV, and XRCC4) in 254 primary breast cancer patients and 379 healthy controls. Support for these hypotheses came from the observations that (a) two SNPs in Ku70 and XRCC4 were associated with breast cancer risk (P < 0.05); (b) a trend toward increased risk of developing breast cancer was found in women harboring a greater number of putative high-risk genotypes of NHEJ genes (an adjusted odds ratio of 1.46 for having one additional putative high-risk genotype; 95% confidence interval, 1.19-1.80); (c) this association between risk and the number of putative high-risk genotypes was stronger and more significant in women thought to be more susceptible to estrogen, i.e., those with no history of full-term pregnancy; and (d) the protective effect conferred by a history of full-term pregnancy was only significant in women with a lower number of putative high-risk genotypes of NHEJ genes. Based on comprehensive NHEJ gene profiles, this study provides new insights to suggest the role of the NHEJ pathway in breast cancer development and supports the possibility that breast cancer is initiated by estrogen exposure, which causes DNA DSBs. PMID- 12750265 TI - Therapeutic targeting of the endothelin a receptor in human ovarian carcinoma. AB - The endothelin A receptor (ET(A)R) autocrine pathway is overexpressed in many malignancies, including ovarian carcinoma. In this tumor, engagement of ET(A)R triggers tumor growth, survival, neoangiogenesis, and invasion. To evaluate whether ET(A)R represents a new target in cancer treatment, we examine in vitro and in vivo the effect of the selective ET(A)R antagonist ABT-627 (atrasentan), a small p.o. bioavailable molecule, in mono- and combination therapy with taxane. ABT-627 effectively inhibits cell proliferation, vascular endothelial growth factor (VEGF) secretion of ovarian carcinoma cell lines, and primary cultures. ET(A)R blockade also results in the sensitization to paclitaxel-induced apoptosis. In ovarian carcinoma xenografts, in which the ET-1/ET(A)R autocrine pathway is overexpressed, tumor growth was significantly inhibited in ABT-627 treated mice compared with control. The therapeutic efficacy of ABT-627 was associated with a significant reduction in microvessel density, expression of VEGF, and matrix metalloproteinase-2, and increased the percentage of apoptotic tumor cells. Combined treatment of ABT-627 with paclitaxel produced additive antitumor, apoptotic, and antiangiogenic effects. These findings demonstrate that the small molecule ABT-627 is a candidate for clinical testing as an antitumor agent in ovarian cancer patients, especially in combination with taxane therapy. Interruption of ET(A)R signaling therefore, represents, a promising therapeutic strategy in ovarian carcinoma. PMID- 12750266 TI - Synergistic down-regulation of urokinase plasminogen activator receptor and matrix metalloproteinase-9 in SNB19 glioblastoma cells efficiently inhibits glioma cell invasion, angiogenesis, and tumor growth. AB - The binding of urokinase plasminogen activator (uPA) to its receptor (uPAR) initiates a proteolytic cascade facilitating the activation of matrix metalloproteinase-9 (MMP-9), which in turn degrades the extracellular matrix. These processes have an established role in tumor invasion and metastasis. Our previous work revealed an inverse association between glioma invasion and the expression of uPAR and MMP-9. In the present study, we used the adenovirus serotype 5 vector system to generate a replication-deficient recombinant adenovirus capable of simultaneously expressing antisense uPAR and antisense MMP 9 (Ad-uPAR-MMP-9). This adenoviral construct is driven by the independent promoter elements cytomegalovirus and bovine growth hormone and SV40 polyadenylation signals to down-regulate key steps in the proteolytic cascade. Ad uPAR-MMP-9 infection of SNB19 cells significantly decreased uPAR and MMP-9 expression as determined by immunohistochemical and Western blotting analyses. A Matrigel invasion assay revealed marked reduction in the invasiveness of the Ad uPAR-MMP-9-infected cells compared with parental and vector controls. Tumor spheroids infected with Ad-uPAR-MMP-9 and cocultured with fetal rat brain aggregates did not invade rat brain aggregates, whereas 90-95% of the mock and empty vector-infected cells invaded the rat brain aggregates. Intracranial injection of SNB19 cells infected ex vivo with the Ad-uPAR-MMP-9 antisense bicistronic construct showed decreased invasiveness and tumorigenicity. s.c. injections of the bicistronic antisense construct into established tumors (U87 MG) caused tumor regression. These results support the therapeutic potential of targeting the individual components of the uPAR-MMP-9 by using a single adenovirus construct for the treatment of gliomas and other cancers. PMID- 12750267 TI - Use of a vaccine strain of measles virus genetically engineered to produce carcinoembryonic antigen as a novel therapeutic agent against glioblastoma multiforme. AB - Despite the most aggressive medical and surgical treatments, glioblastoma multiforme remains incurable with a median survival of <1 year. We investigated the antitumor potential of a novel viral agent, an attenuated strain of measles virus (MV), derived from the Edmonston vaccine lineage, genetically engineered to produce carcinoembryonic antigen (CEA). CEA production as the virus replicates can serve as a marker of viral gene expression. Infection of a variety of glioblastoma cell lines including U87, U118, and U251 at MOIs 0.1, 1, and 10 resulted in significant cytopathic effect consisting of excessive syncycial formation and massive cell death at 72-96 h from infection. terminal deoxynucleotidyltransferase-mediated nick end labeling assays demonstrated the mechanism of cell death to be predominantly apoptotic. The efficacy of this approach in vivo was examined in BALB/c nude mice by using both s.c. and intracranial orthotopic U87 tumor models. In the s.c. U87 model, mice with established xenografts were treated with a total dose of 8 x 10(7) plaque forming units of MV-CEA, administered i.v. Mice treated with UV light inactivated MV, and untreated mice with established U87 tumors were used as controls. There was statistically significant regression of s.c. tumors (P < 0.001) and prolongation of survival (P = 0.007) in MV-CEA treated animals compared with the two control groups. In the intracranial orthotopic U87 model, there was significant regression of intracranial U87 tumors treated with intratumoral administration of MV-CEA at a total dose of 1.8 x 10(6) plaque forming units as assessed by magnetic resonance image (P = 0.002), and statistically significant prolongation of survival as compared with mice that received UV-inactivated virus and untreated mice (P = 0.02). Histological examination of brains of MV-CEA-treated animals revealed complete regression of the tumor with the presence of a residual glial scar and reactive changes, mainly presence of hemosiderin-laden macrophages. In addition, CEA levels in the peripheral blood in both the s.c. and orthotopic models increased before tumor regression, indicating viral gene expression, and returned to normal when the tumors regressed. Ifnar(ko) CD46 Ge transgenic mice, susceptible to MV infection, were used to assess central nervous system toxicity of MV-CEA. Intracranial administration of MV-CEA into the caudate nucleus of Ifnar(ko) CD46 Ge did not result in clinical neurotoxicity. Pathologic examination demonstrated limited microglial infiltration surrounding the injection site. In summary, MV-CEA has potent antitumor activity against gliomas in vitro, as well as in both s.c. and orthotopic U87 animal models. Monitoring CEA levels in the serum can serve as a low-risk method of detecting viral gene expression during treatment, and could allow dose optimization and individualization of treatment. PMID- 12750268 TI - Immuno-gene therapy of established prostate tumors using chimeric receptor redirected human lymphocytes. AB - Targeted adoptive immunotherapy is an attractive option for prostate cancer given its accessible primary location, the presence of specific tissue and tumor antigens, and the acceptability of collateral destruction of healthy prostrate tissue. The "T-body" approach, which uses genetically programmed, patient-derived lymphocytes transfected with chimeric receptor genes, combines the effector functions of T lymphocytes and natural killer cells with the ability of antibodies to recognize predefined surface antigens with high specificity and in a non-MHC restricted manner. We evaluated the therapeutic efficacy of anti-erbB2 chimeric receptor-bearing human lymphocytes on human prostate cancer xenografts in a SCID mouse model. Local delivery of erbB2-specific T bodies to well established s.c. and orthotopic tumors, together with systemic administration of interleukin-2, resulted in retardation of both tumor growth and prostate-specific antigen secretion, prolongation of survival, and complete tumor elimination in a significant number of mice. These preclinical studies demonstrate the therapeutic potential of the T-body approach for locally advanced or recurrent prostate cancer as an adjunct to, or after, conventional therapy. PMID- 12750269 TI - High-resolution magnetic resonance imaging of the efficacy of the cytosine analogue 1-[2-C-cyano-2-deoxy-beta-D-arabino-pentofuranosyl]-N(4)-palmitoyl cytosine (CS-682) in a liver-metastasis athymic nude mouse model. AB - High-resolution magnetic resonance (MR) imaging techniques in a liver metastatic mouse model were used to assess CS-682, a novel 2'-deoxycytidine analogue of 1-[2 C-cyano-2-deoxy-beta-D-arabino-pentofuranosyl]-N(4)-palmitoyl cytosine. The efficacy of CS-682 was visualized in real time by MR imaging of initial seeding and subsequent growth of liver metastases. The relative therapeutic efficacies of CS-682 and two agents used clinically, gemcitabine [2'-deoxy-2',2' difluorocytidine monohydrochloride (DFDC)] and 5-fluorouracil (5-FU), were compared in this model. CS-682 was found to exhibit superior efficacy by delaying the onset and inhibiting the growth of liver metastasis compared with gemcitabine, 5-FU, and control. The overall occurrence of metastases was decreased 62% by CS-682, 18% by DFDC, and 35% by 5-FU. CS-682 increased the life span of the treated animals significantly, by 28 days above the 29-day median survival without treatment, compared with 11 days by DFDC and 14 days by 5-FU. The increased survival in CS-682-treated animals correlated with the antimetastatic activity of this compound. These preclinical findings support the potential clinical utility of CS-682 in the treatment of liver metastasis. PMID- 12750270 TI - RRR-alpha-tocopheryl succinate-induced apoptosis of human breast cancer cells involves Bax translocation to mitochondria. AB - Previous studies have identified RRR-alpha-tocopheryl succinate (vitamin E succinate, VES) as a potential chemotherapeutic agent. VES induces human breast cancer cells to undergo apoptosis in a concentration- and time-dependent manner by restoring transforming growth factor beta (TGF-beta) and Fas (CD95) apoptotic signaling pathways, that contribute to the activation of c-Jun NH(2)-terminal kinase (JNK)-mediated apoptosis. The objective of these studies was to clarify biochemical events involved in VES-induced apoptosis. Data show that VES-induced apoptosis involves: (a) translocation of Bax from the cytosol to the mitochondria and cytochrome c release from the mitochondria to the cytosol as determined by Western immunoblot analyses of mitochondrial- and cytosolic-enriched cellular fractions; (b) increased permeabilization of mitochondrial membranes as determined by confocal and fluorescence-activated cell sorting analyses of loss of a mitochondrial selective fluorescent dye; (c) processing of caspase-9 and -3 but not caspase-8 to active forms and cleavage of poly(ADP-ribose) polymerase (PARP) as determined by Western immunoblot analyses using antibodies capable of detecting both proenzyme and processed enzyme forms or the intact or cleaved forms of PARP. Transient transfection of cells with antisense oligonucleotides to Bax or transient overexpression of Bcl-2 prevented VES-induced mitochondrial permeability transition and apoptosis. The use of cell-permeable caspase inhibitors indicated that caspase-9 and -3 but not caspase-8 are involved in VES induced apoptosis. JNK inhibitor II blocked VES-induced Bax conformational change, indicating a role for JNK in Bax translocation to the mitochondria. Taken together, these data suggest that the activation of JNK, translocation of Bax to the mitochondria, increased mitochondrial membrane permeability with release of cytochrome c, and activation of caspase-9 and -3 are critical events in VES induced apoptosis of human MDA-MB-435 breast cancer cells. PMID- 12750271 TI - Effect of the multidrug resistance protein on the transport of the antiandrogen flutamide. AB - Prostate cancer is the most common noncutaneous malignancy of American men. Although it can be initially treated with androgen deprivation therapy, tumors that relapse become resistant to future hormonal manipulation. We previously found that the multidrug resistance protein (MRP), MRP1, is overexpressed in advanced stage and grade human prostate cancer and is negatively regulated by p53. In this study, we sought to determine whether the cellular accumulation of the antiandrogen flutamide, a drug commonly used in the treatment of prostate cancer, is affected by MRP1 expression. There were significant differences between the wild-type and MRP1-overexpressing cells in efflux and accumulation of flutamide and hydroxyflutamide, its active metabolite. In contrast, transport of dihydrotestosterone was not affected by MRP1. Treating the cells with leukotriene D4, a known MRP1 substrate, or VX-710, an MRP1 modulator, restored flutamide and hydroxyflutamide accumulation. Finally, intracellular glutathione depletion with buthionine sulfoximine or energy depletion using 2-deoxy-D-glucose/sodium azide restored flutamide accumulation to that of parental cells while incubating the cells at 4 degrees C abolished MRP1-mediated transport. In summary, these studies indicate that flutamide and hydroxyflutamide but not dihydrotestosterone are transported by MRP1 and that these findings may contribute to our understanding of resistance to hormone refractory prostate cancer. PMID- 12750273 TI - Identification of the human IAI.3B promoter element and its use in the construction of a replication-selective adenovirus for ovarian cancer therapy. AB - Little is known concerning promoters or gene therapy specific for ovarian cancer. To explore the potential use of IAI.3B isolated from ovarian cancer cells in gene therapy for ovarian cancer, we identified the promoter region of the IAI.3B gene and created a replication-selective adenovirus, AdE3-IAI.3B, driven by the promoter. Transient transfection experiments showed that the DNA segment located between -1816 and -1 bp resulted in preferential expression in ovarian cancer cells with negligible expression in squamous cell carcinoma and normal cells. The promoter activity of IAI.3B was almost the same as that of cytomegalovirus and an order of magnitude higher than those of midkine and cyclooxygenase-2 in ovarian cancer cells. AdE3-IAI.3B replicated as efficiently as the wild-type adenovirus and caused extensive cell killing in a panel of ovarian cancer cells in vitro. In contrast, squamous cell carcinoma and normal cells were not able to support AdE3 IAI.3B replication. In animal studies, AdE3-IAI.3B administered to flank and i.p. xenografts of ovarian cancer cells led to a significant therapeutic effect. These results demonstrate the usefulness of the IAI.3B promoter for generation of ovarian cancer-specific adenoviral vectors and provide a potential for the development of ovarian cancer-specific oncolytic viral therapies. PMID- 12750272 TI - Local intracerebral delivery of endogenous inhibitors by osmotic minipumps effectively suppresses glioma growth in vivo. AB - The systemic administration of endogenous inhibitors significantly reduced the growth of human glioma in vivo, but required the production of a large amount of biologically active protein. In this study we reduced the amount of protein needed and optimized the therapeutical response by delivering the endogenous inhibitors locally into the brain by osmotic minipumps. Human hemopexin fragment of MMP-2 or COOH-terminal fragment of platelet factor-4 were delivered locally and continuously into the brain of mice implanted intracranially with glioma cells, by osmotic minipumps connected to an intracranial catheter. Local delivery of human hemopexin fragment of MMP-2 and COOH-terminal fragment of platelet factor-4 significantly inhibited the growth of well-established malignant glioma in nude and BALB/C mice. When the inhibitors were given at the same concentration, the efficacy of the local delivery was much higher than that reached with the systemic administration, both when the inhibitor was administered daily or continuously by s.c. minipumps. Moreover, the local delivery reduced the amount of protein needed to reach a significant therapeutic response. Intracerebral delivery maintained a long-term control of glioma growth and inhibited glioma recurrence in a surgical resection model. Treatment showed no side effects. Histochemical analysis of tumors showed that the tumor growth inhibition was the result of a decrease in tumor vasculature and a change in tumor vessel morphology. Our data demonstrate that local intracerebral delivery of endogenous inhibitors effectively inhibits malignant glioma growth and reduces the amount of protein needed to reach a therapeutical response. PMID- 12750274 TI - Enhancement of radiation response by roscovitine in human breast carcinoma in vitro and in vivo. AB - Frequent deregulation of cyclin-dependent kinase (CDK) activation associated with loss of cell cycle control was found in most of human cancers. A recent development of a new class of antineoplasic agents targeting the cell cycle emerged as a small molecule CDK inhibitor, roscovitine, which presents potential antiproliferative and antitumoral effects in human tumors. Additional studies reported that roscovitine combined with cytotoxic agents can cooperate with DNA damage to activate p53 protein. However, little is known about the biological effect of roscovitine combined with ionizing radiation (IR) in human carcinoma, and no studies were reported thus far in p53 mutated carcinoma. In the breast cancer cell line MDA-MB 231, which lacks a functional p53 protein, we found a strong radiosensitization effect of roscovitine in vitro by clonogenic survival assay and in vivo in MDA-MB 231 xenograft model. Using Pulse Field Gel Electrophoresis, a strong impairment in DNA-double-strand break rejoining was observed after roscovitine and IR treatment as compared with IR alone. Cell cycle analysis showed a G(2) delay and no increase in radiation-induced apoptosis in the cells treated with IR or roscovitine and IR. On the other hand, we found a significant induction in micronuclei frequency after roscovitine and IR treatment as compared with IR alone. This effect was also observed in BALB murine cells in contrast to SCID murine cells, which are deficient in DNA-PKcs, suggesting a possible DNA-double-strand break repair defect in the nonhomologous end joining pathways. In MDA-MB 231 cells, the radiosensitization effect of roscovitine was associated with an inhibition of the DNA-dependent protein kinase activity caused by a marked decrease in Ku-DNA binding by using the electrophoretic mobility shift assay. In conclusion, we found a novel effect on DNA repair of the CDK inhibitor roscovitine, which acts as a radiosensitizer in vitro and in vivo in breast cancer cells lacking a functional p53. PMID- 12750275 TI - LAG-3 enables DNA vaccination to persistently prevent mammary carcinogenesis in HER-2/neu transgenic BALB/c mice. AB - Within 33 weeks of life, all 10 mammary glands of virgin BALB/c mice transgenic for the transforming rat HER-2/neu oncogene under the mammary tumor virus promoter (BALB-neuT mice) progress from atypical hyperplasia to invasive palpable carcinoma. Repeated DNA vaccination with plasmids coding for the extracellular and transmembrane domain of the protein product of rat HER-2/neu (r-p185(neu)) delayed tumor onset and reduced tumor multiplicity, but this protection eventually declined, and few mice were tumor free at 1 year of age. Association of plasmid vaccination with administration of soluble mouse LAG-3 (lymphocyte activation gene-3/CD223) generated by fusing the extracellular domain of murine LAG-3 to a murine IgG2a Fc portion (mLAG-3Ig) elicited a stronger and sustained protection that kept 70% of 1-year-old mice tumor free. Moreover, this combined vaccination, which was performed when multiple in situ carcinomas were already evident, extended disease-free survival and reduced carcinoma multiplicity. Inhibition of carcinogenesis was associated with markedly reduced epithelial cell proliferation and r-p185(neu) expression, whereas the few remaining hyperplastic foci were heavily infiltrated by reactive leukocytes. A stronger and enduring r p185(neu)-specific cytotoxicity, a sustained release of IFN-gamma and interleukin 4, and a marked expansion of both CD8(+)/CD11b(+)/CD28(+) effector and CD8(+)/CD11b(+)/CD28(-) memory effector T-cell populations were induced in immunized mice. This combined vaccination also elicited a quicker and higher antibody response to r-p185(neu), as well as an early antibody isotype switch. These data suggest that the appropriate costimulation provided by mLAG-3Ig enables DNA vaccination to establish an effective protection, probably by enhancing cross-presentation of the DNA coded antigen. PMID- 12750276 TI - Archaeosomes induce enhanced cytotoxic T lymphocyte responses to entrapped soluble protein in the absence of interleukin 12 and protect against tumor challenge. AB - Archaeosome adjuvants formulated as archaeal ether glycerolipid vesicles induce strong CD4(+) as well as CD8(+) CTL responses to entrapped soluble antigens. Immunization of mice with ovalbumin (OVA) entrapped in archaeosomes composed of the total polar lipids of Methanobrevibacter smithii resulted in a potent OVA specific CD8(+) T-cell response, and subsequently, the mice dramatically resisted solid tumor growth of OVA-expressing EG.7 cells and lung metastasis of B16OVA melanoma cells. Prophylactic protection was antigen-specific because tumor curtailment was not seen in mice injected with antigen-free archaeosomes. Similarly, there was no protection against B16 melanoma cells lacking OVA expression. Furthermore, in vivo depletion of CD8(+) T cells abrogated the protective response, indicating that the antitumor immunity was mediated by CTLs. Depletion of CD4(+) T cells also resulted in partial loss of tumor protection, suggesting a beneficial role for T-helper cells. Interestingly OVA-archaeosomes induced enhanced CTL response in the absence of interleukin 12 and IFN-gamma. Furthermore, interleukin 12-deficient mice mounted strong tumor protection. However, IFN-gamma-deficient mice, despite the strong CTL response, were only transiently protected, revealing a need for IFN-gamma response in tumor protection. Archaeosomes also facilitated therapeutic protection when injected into mice concurrent with tumor cells. Interestingly, even archaeosomes lacking entrapped antigen mediated therapeutic protection, and this correlated to the activation of innate immunity as evident by the increased tumor-infiltrating natural killer and dendritic cells. Thus, archaeosomes represent effective tumor antigen delivery vehicles that can mediate protection by activating both innate as well as acquired immunity. PMID- 12750277 TI - Lack of terminally differentiated tumor-specific CD8+ T cells at tumor site in spite of antitumor immunity to self-antigens in human metastatic melanoma. AB - Activation of CTL-mediated antitumor immunity to self-epitopes expressed by neoplastic cells is thought to be prevented, at any stage of tumor progression, by tolerance mechanisms. In contrast, in 74 American Joint Committee on Cancer stages I-IV melanoma patients, we found that development of lymph node metastases is a key event triggering CD8(+) T-cell-mediated immunity to self-epitopes encoded by melanocyte differentiation antigens. This was shown by the increased peripheral precursor frequency to Melan-A/Mart-1, gp100, and tyrosinase epitopes in stage III and IV compared with stage I and II patients, and by accumulation of functional memory T cells directed to Melan-A/Mart-1(26-35) in tumor-invaded lymph nodes. However, in tumor-invaded lymph nodes of most patients, CD8(+) T cells directed to melanocyte differentiation antigens or to tumor-restricted antigens (MAGE-3 and NY-ESO-1 epitopes), showed a CCR7(+) CD45RA(+) CD27(+) CD28(+) perforin(-) "precursor" phenotype. Only in 7 of 23 cases antigen-specific CD8(+) T cells in invaded lymph nodes showed a predominant CCR7(-) CD45RA(-) CD27(+) CD28(-) perforin(+) "preterminally differentiated" phenotype. In the latter subset of patients, by immunohistochemistry in lymph node lesions, we found that CD8(+) T lymphocytes intermingling with the neoplastic tissue expressed a CCR7(-) CD45RO(+)/RA(-) phenotype, whereas CD4(+) lymphocytes did not infiltrate the tumor. Furthermore, perforin and granzyme B were expressed on a higher fraction of the CD8(+) cells surrounding the invading tumor compared with the lymphocytes infiltrating the neoplastic tissue. In addition, no evidence for tumor regression was found in such metastatic lesions, as documented by absence of neoplastic cell necrosis or apoptosis. These data indicate that neoplastic cells in the lymph nodes and/or increased tumor burden in metastatic disease activate CD8(+) T-cell-mediated antitumor immunity to self-epitopes. However, the paucity of terminally differentiated CD8(+) T cells at tumor site suggests that immunotherapy strategies may require not only the boosting of tumor immunity, but also effective means to promote CD8(+) T-cell differentiation in the neoplastic tissue. PMID- 12750278 TI - Polarization effects of 4-1BB during CD28 costimulation in generating tumor reactive T cells for cancer immunotherapy. AB - Using murine tumor-draining lymph node (TDLN) cells, we investigated the polarization effect of 4-1BB (CD137) during CD28 costimulation in generating antitumor T cells. Costimulation of TDLN cells through the newly induced 4-1BB molecules, CD3, and CD28 using monoclonal antibodies significantly enhanced cell proliferation. The greater cell yield with 4-1BB signaling appeared to be related to the inhibition of activation-induced cell death. Activation of TDLN cells through 4-1BB in addition to CD3/CD28 signaling shifted T-cell responses toward a type 1 cytokine pattern because 4-1BB ligation plus CD3/CD28 stimulation significantly augmented type 1 cytokine (e.g., IFN-gamma) and granulocyte macrophage colony-stimulating factor secretion. By contrast, type 2 cytokine (e.g., interleukin 10) secretion by the activated TDLN cells was significantly reduced. The in vivo antitumor reactivity of TDLN cells activated through 4-1BB in conjunction with CD3/CD28 pathways was examined using an adoptive immunotherapy model. The number of pulmonary metastases was significantly reduced and survival was prolonged after the transfer of anti-CD3/anti-CD28/anti-4-1BB activated TDLN cells compared with an equivalent number of cells activated without anti-4-1BB. The antitumor effect through 4-1BB involvement during CD28 costimulation was dependent on IFN-gamma production and abrogated after IFN-gamma neutralization. By contrast, interleukin 10 neutralization resulted in significantly enhanced tumor regression. These results indicate that costimulation of TDLN cells through newly induced 4-1BB and CD3/CD28 signaling can significantly increase antitumor reactivity by shifting T-cell responses toward a type 1 cytokine pattern while concomitantly decreasing type 2 responses. PMID- 12750279 TI - Targeted immunotherapy using reconstituted chaperone complexes of heat shock protein 110 and melanoma-associated antigen gp100. AB - This report defines a novel approach to heat shock protein vaccine formulation that takes advantage of the chaperoning property of heat shock protein hsp110 to efficiently bind a large protein substrate (specifically, human melanoma associated antigen gp100) during heat shock. We demonstrate that hsp110 can form chaperone complexes with gp100 and prevent heat-induced aggregation of gp100. The resultant natural hsp110-gp100 complexes are strongly immunogenic as determined by their ability to elicit an antigen-specific IFN-gamma production and a cytotoxic T-cell response. Immunization with the hsp110-gp100 complex protected mice against subsequent challenge with human gp100-transduced B16 melanoma, which involves both CD4(+) and CD8(+) T-cell populations. Administration of the hsp110 gp100 vaccine also significantly suppressed the growth of established tumors in a therapeutic model. Furthermore, the hsp110-gp100 chaperone complex exhibited inhibitory effects on the progression of wild-type B16 tumor, suggesting that the induced immune response by human gp100 cross-reacts with mouse gp100. More importantly, the antitumor response obtained with the hsp110-gp100 complex is more potent than that obtained using Complete Freund's Adjuvant with gp100, whereas no response was observed against mouse hsp110 itself. Thus, the use of hsp110 to form natural chaperone complexes with tumor protein antigens such as gp100 represents a powerful approach to therapeutic vaccine formulation with significant potential for clinical application. PMID- 12750280 TI - Expansion of tumor-specific CD8+ T cell clones in patients with relapsed myeloma after donor lymphocyte infusion. AB - Donor lymphocyte infusions (DLI) provide effective therapy for patients with multiple myeloma who have relapsed after allogeneic bone marrow transplantation. However, the immunological mechanisms of the graft-versus-myeloma (GVM) effect have not been defined, and the target antigens of this response have not been identified. Molecular analysis of CDR3 Vbeta repertoire after CD4+ DLI demonstrated previously that the development of GVM and graft-versus-host-disease (GVHD) were associated with the clonal expansion of distinct T-cell populations in patient peripheral blood. In the current study, we undertook a molecular and functional characterization of GVM- and GVHD-associated T-cell clones. T-cell clones associated with GVM were detectable by clone-specific PCR at a low level in peripheral blood before DLI and expanded approximately 10-fold after DLI. In contrast, T-cell clones associated with GVHD were not detectable before DLI or before the development of clinical GVHD. Two T-cell clones associated with GVM were isolated and expanded in vitro, allowing their phenotypic and functional characterization. Both GVM clones were derived from donor cells and had a CD3+CD8+CD4- phenotype. One GVM clone specifically recognized patient myeloma cells in an HLA class I-restricted manner, but was not reactive with patient normal bone marrow cells or patient EBV transformed B cells. Taken together, these findings suggest that the GVM response is mediated by donor-derived CD8+ T cell clones with antimyeloma specificity that may be present before DLI. In contrast, T-cell clones associated with GVHD are expanded de novo after DLI. PMID- 12750281 TI - Distinctive patterns of gene expression in premalignant gastric mucosa and gastric cancer. AB - Current epidemiological evidence supports a pathogenetic model of gastric cancer involving intermediate stages that include chronic gastritis and intestinal metaplasia. This study explores the molecular features of gastric cancer and premalignant stages using DNA microarray-based gene expression profiling and relates these findings to clinical, pathological, and ethnic parameters. A total of 124 tumor and adjacent mucosa samples were analyzed using spotted cDNA microarrays containing 9381 nonredundant gene elements. Tumor specimens were diffuse, intestinal, or mixed gastric cancer and adjacent mucosa, which generally displayed signs of chronic gastritis or intestinal metaplasia. Expression patterns could be discerned that readily defined premalignant and tumor subtypes. Chronic gastritis exhibits a pronounced mitochondrial gene expression signature, which may be linked to Helicobacter pylori pathogenesis. Intestinal metaplasia was associated with increased expression of many intestinal differentiation genes, many of which were not overexpressed in tumors. Samples were obtained from 91 Australian and 33 Chinese patients to explore potential variation in gene expression between these populations. Despite differences in the incidence, and potentially the etiology, of gastric cancer between these ethnic groups, we found the tumors to be molecularly similar. The identification of molecular signatures that are characteristic of subtypes of gastric cancer and associated premalignant changes should enable further analysis of the steps involved in the initiation and progression of this disease. PMID- 12750282 TI - Tumor classification based on gene expression profiling shows that uveal melanomas with and without monosomy 3 represent two distinct entities. AB - Uveal melanoma is the most common intraocular malignancy. About 50% of patients die of metastases, which almost exclusively originate from primary tumors that have lost one chromosome 3 (monosomy 3). To gain insight into the biological mechanisms that underlie the various metastasizing potential of uveal melanoma, we have determined gene expression levels in 20 primary tumors using oligonucleotide microarrays containing 12500 probe sets. The expression measurements of those 7902 genes that were expressed in more than 10% of tumors were analyzed using two different statistical approaches. We used a modified Wilcoxon rank-sum test to identify genes differentially expressed between tumors with and without monosomy 3. Seven genes showed complete loss of expression in tumors with monosomy 3 but were expressed in tumors with disomy 3. Two of them, CHL1 and fls485, are located within or close to the uveal melanoma susceptibility locus UVM2 at 3p25. However, mutation analysis of both genes in eight tumors with monosomy 3 did not reveal structural or epigenetic alteration. To identify tumor classes, we performed unsupervised hierarchical cluster analysis; this approach separated uveal melanomas into two groups. We found that this classification is strikingly robust because, when tested by "resampling," the same grouping is obtained from 47 of 50 subsamples of genes. In clusterings of the three remaining subsamples, the grouping of only one tumor does not conform with the original classification. Excluding this tumor, cluster analyses of subsamples containing as few as 300 randomly chosen genes consistently result in the same classification, thus indicating that the difference between the two tumor classes is pervasive. Interestingly, all of the tumors in one of the groups have disomy 3, whereas all of the others have monosomy 3. Our findings suggest that there are two distinct entities of uveal melanoma that were previously unrecognized because they are not obviously distinguishable by clinicopathological features. PMID- 12750283 TI - Fanconi anemia gene mutations in young-onset pancreatic cancer. AB - Genes of the Fanconi complementation groups [Fanconi anemia (FA) genes] are suggested to be involved in homologous DNA recombination and produce FA when two allelic mutations are inherited. BRCA2 is an FA gene and additionally conveys an inherited risk for breast, ovarian, and pancreatic cancer for individuals carrying a single mutated allele [N. G. Howlett et al., Science (Wash. DC), 297: 606-609, 2002]. Here we report inherited and somatic mutations of FANCC and FANCG present in young-onset pancreatic cancer. This may imply a general involvement of Fanconi genes with an inherited risk of cancer. The known hypersensitivity of Fanconi cells to mitomycin and other therapeutic agents [M. S. Sasaki, Nature (Lond.), 257: 501-503, 1975] suggests a therapeutic utility for a more complete characterization of the DNA repair defects and their causative genetic mutations in pancreatic cancer. PMID- 12750284 TI - Assembly of functional ALT-associated promyelocytic leukemia bodies requires Nijmegen Breakage Syndrome 1. AB - Immortalized cells maintain telomere length through either a telomerase-dependent process or a telomerase-independent pathway termed alternative lengthening of telomeres (ALT). Homologous recombination is implicated in the ALT pathway in both yeast and human ALT cells. In ALT cells, two types of DNA double-strand break repair and homologous recombination factors, the Rad50/Mre11/NBS1 complex and Rad51/Rad52 along with replication factors (RPA) and telomere binding proteins (TRF1 and TRF2), are associated with the ALT-associated PML body (APB). DNA synthesis in late S-G(2) is associated with APBs, which contain telomeric DNA and, are therefore, potential sites for telomere length maintenance. Here, we show that the breast cancer susceptibility gene product, breast cancer susceptibility gene 1, and the human homologue of yeast Rap1, hRap1, are also associated with APBs specifically during late S-G(2) phase of the cell cycle. We additionally show that the localization of the double-strand break repair factors with APBs is distinct from their association with ionizing radiation-induced nuclear foci. To systematically explore the mechanism involved in the assembly of APBs, we examine the role of Nijmegen breakage syndrome 1 (NBS1) and TRF1 in this process, respectively. We demonstrated that NBS1 plays a key role in the assembly and/or recruitment of Rad50, Mre11, and breast cancer susceptibility gene 1, but not Rad51 or TRF1, to APBs. The NH(2) terminus of NBS1, specifically the BRCA1 COOH-terminal domain, is required for this activity. Although TRF1 interacts with NBS1 directly, it is dispensable for the association of either Rad50/Mre11/NBS1 or Rad51 with APBs. Perturbation of the interactions between NBS1/Mre11 and APBs correlates with reduced BrdUrd incorporation associated with APBs, consistent with decreased DNA synthesis at these sites. Taken together, these results support a model in which NBS1 has a vital role in the assembly of APBs, which function to maintain telomeres in human ALT cells. PMID- 12750285 TI - p53 interacts with hRAD51 and hRAD54, and directly modulates homologous recombination. AB - p53 inhibits tumorigenesis through a variety of functions, including mediation of cell cycle arrest, premature senescence, and apoptosis.p53 also can associate with several DNA helicases and proteins involved in homologous recombination. In this study, we show that p53, hRAD51, and hRAD54 coimmunoprecipitated and colocalized with each other at endogenous levels in normal cells. Colocalization was observed with the phosphoserine-15 form of p53 at presumed DNA processing sites after the induction of DNA breaks. hRAD54 bound directly to the p53 COOH terminus in vitro without a nucleic acid intermediate. We then investigated the functional consequences of these protein interactions. A host cell reactivation assay revealed that the elevation in recombination observed after p53 inactivation is dependent on the hRAD51 pathway and that p53-dependent antirecombinogenic activity can be attributed to p53 binding to hRAD51 directly. These data support the hypothesis that p53 helps maintain genetic stability through transcription-independent modulation of homologous recombination factors. PMID- 12750286 TI - Allelic losses on chromosome 6q25 in Hodgkin and Reed Sternberg cells. AB - We established a molecular cytogenetic approach to identify consistent genetic aberrations in classical Hodgkin lymphoma. Single laser-micromanipulated Hodgkin and Reed Sternberg (H-RS) cells and the respective germ line tissue were PCR amplified using highly polymorphic microsatellite probes. Loss of heterozygosity and genomic imbalances of the fluorochrome-labeled microsatellites were determined by fragment length analysis. Eleven cases of in classical Hodgkin lymphoma (cHL) were initially screened with 21 microsatellite markers scattered over the entire genome. Loss of heterozygosity was detected in >40% of informative loci in most cases indicating a deletion of a substantial part of the genome of H-RS cells. Allelic losses and imbalances on chromosome 6q were detected in most of these cases. A deletion mapping of 6q was performed in 16 cases of cHL. This detailed analysis of 6q led to the identification of a 3.3-Mbp region around D6S311 flanked by D6S978 and D6S1564 that was altered in 11 of 14 cases of cHL analyzed. In conclusion, allelotyping of single H-RS cells revealed monoallelic chromosomal deletions and genomic imbalances on 6q that might affect genes critically involved in the pathogenesis of H-RS cells. PMID- 12750287 TI - A transforming growth factorbeta1 signal peptide variant increases secretion in vitro and is associated with increased incidence of invasive breast cancer. AB - There is evidence that transforming growth factor (TGF)beta acts as a suppressor of tumor initiation but also as a promoter of tumor progression when the antiproliferative effect of the TGFbeta signaling pathway has been overridden by other oncogenic mutations. Several somatic mutations that disrupt the TGFbeta SMAD signaling pathway have been reported in human breast tumors. We have examined the association between single nucleotide polymorphisms (SNPs) in the TGFbeta1 gene and the incidence of invasive breast cancer in three case-control series, with a maximum of 3987 patients and 3867 controls, median age approximately 50 years, and range 22-92 years. The promoter SNP, C-509T, and the T +29C signal-peptide SNP (encoding Leu10Pro) are in strong linkage disequilibrium. They are both significantly associated with increased incidence of invasive breast cancer in a recessive manner [odds ratios: (TT versus C carrier), 1.25; 95% confidence intervals 1.06-1.48; P = 0.009 and (ProPro versus Leu-carrier), 1.21; 95% confidence intervals 1.05-1.37; P = 0.01]. The G-800A SNP was not significantly associated with incidence of breast cancer. The C-509T SNP is not contained within a known consensus sequence for a promoter regulatory element and therefore unlikely to affect TGFbeta1 expression, whereas the Leu10Pro signal peptide substitution potentially affects TGFbeta1 secretion. Transfections of HeLa cells with constructs encoding either the Pro or Leu forms of TGFbeta1 and driven by the cytomegalovirus promoter indicate that the signal peptide with Pro at residue 10 causes a 2.8-fold increase in secretion compared with the Leu form. These data indicate that the allele encoding Pro10 is associated with increased rates of TGFbeta1 secretion and with increased incidence of invasive breast cancer for the population samples described. It is estimated that 3% of all breast cancer cases may be attributable to Pro10 homozygosity. PMID- 12750288 TI - p53-independent activation of the hdm2-P2 promoter through multiple transcription factor response elements results in elevated hdm2 expression in estrogen receptor alpha-positive breast cancer cells. AB - The negative-regulatory feedback loop between p53 and hdm2 forms part of a finely balanced regulatory network of proteins that controls cell cycle progression and commitment to apoptosis. Expression of hdm2, and its mouse orthologue mdm2, is known to be induced by p53, but recent evidence has demonstrated mdm2 expression can also be regulated via p53-independent pathways. However the p53 independent mechanisms that control transcription of the human hdm2 gene have not been studied. Differential levels of hdm2 mRNA and protein expression have been reported in several types of human malignancy, including breast cancers in which hdm2 expression correlates with positive estrogen receptor alpha (ERalpha) status. Experimental models have demonstrated that hdm2 overexpression can promote breast cancer development. Here, we show that the elevated level of hdm2 protein in ERalpha(+ve) breast cancer cell lines such as MCF-7 and T47D is because of transcription from the p53-inducible P2 promoter of hdm2. The P2 promoter is inactive in ERalpha(-ve) cell lines such as SKBr3. Hdm2-P2 promoter activity in T47D cells is independent of p53, as well as of known regulators of the mouse mdm2-P2 promoter, including ERalpha and ras-raf-mitogen-activated protein/extracellular signal-regulated kinase (MEK) mitogen-activated protein kinase (MAPK) signaling. We show that hdm2-P2 activity in T47D cells is dependent on the integrity of both an evolutionarily conserved composite binding site for AP1 and ETS family transcription factors (AP1-ETS) and a nonconserved upstream (nnGGGGC)(5) repeat sequence. Lack of hdm2-P2 activity in ERalpha(-ve) cells is shown to be a consequence of reduced transcriptional activation through the AP1 ETS element. Overexpression of ETS2 in SKBr3 cells reconstitutes AP1-ETS element dependent hdm2-P2 promoter activity, resulting in increased levels of hdm2 protein in the cells. Our findings support the hypothesis that the elevated levels of hdm2 expression reported in cancers such as ERalpha(+ve) breast tumors play an important role in the development of these tumors. PMID- 12750289 TI - Requirement of a specific Sp1 site for histone deacetylase-mediated repression of transforming growth factor beta Type II receptor expression in human pancreatic cancer cells. AB - In this study, we demonstrate a novel mechanism by which down-regulation of transforming growth factor beta type II receptor (TbetaRII) is mediated by a histone deacetylase (HDAC) in pancreatic ductal adenocarcinoma (PDAC) cells. Treatment of PDAC cell lines BxPC-3 and MIA PaCa-2 with a specific HDAC inhibitor, trichostatin A (TSA), strongly activates TbetaRII promoter activity and induces TbetaRII expression. The transcriptional activation of TbetaRII by TSA was correlated with a decrease in HDAC activity and an increase in acetylated histone H4 protein. Correspondingly, an increase in the association of TbetaRII promoter with acetylated histone H4 was detected in the TSA-treated cells as determined by a chromatin immunoprecipitation assay. We found that a specific Sp1 site (Sp1C, located at -102 bp relative to the transcription start site) adjacent to an inverted CCAAT box (-83 bp) is required for TSA-mediated activation of the TbetaRII promoter. Furthermore, we determined that HDAC1 complexed with Sp1 in PDAC cells and that TSA treatment interfered with this association. Diminished binding of HDAC1 to the -112 to -65 bp region of the TbetaRII promoter after TSA treatment was confirmed by a DNA affinity precipitation assay. This is the first study to demonstrate the requirement of a specific Sp1 site for TSA-mediated transcriptional activation of TbetaRII. This study further suggests that the specificity of this Sp1 site for HDAC-mediated repression of TbetaRII may involve the interaction of the Sp1-HDAC1 complex with components of the cognate transcriptional regulators that bind to the inverted CCAAT box. PMID- 12750290 TI - Osteoblast-related transcription factors Runx2 (Cbfa1/AML3) and MSX2 mediate the expression of bone sialoprotein in human metastatic breast cancer cells. AB - Human breast cancers are known to preferentially metastasize to skeletal sites, however, the mechanisms that mediate the skeletal preference (orthotropism) of specific types of cancers remains poorly understood. There is a significant clinical correlation between the expression of bone sialoprotein (BSP) and skeletal metastasis of breast cancers. Our laboratory, as well as others, have proposed the concept that skeletal selective metastasis and associated disease may be attributable to a mimicry of skeletal cellular phenotypes by metastasizing cancer cells. We hypothesize that breast cancer cell expression of phenotypic properties of skeletal cell types, including BSP as one component of that phenotype, is the result of ectopic expression or activity of one or more central transcriptional regulators of bone cell gene expression. To test this hypothesis, we examined the molecular mechanisms that regulate bsp expression in human breast cancer cell lines with previously characterized metastatic potentials. Our results demonstrate that the majority of the distal bsp promoter sequences act to repress BSP expression in cancer cells and that most of the promoter activity resides in the proximal -110 bp of the bsp promoter. In this region, we identified a putative Runx binding element providing a basis for a mechanism for skeletal gene activation. Our results demonstrate that Runx2 is ectopically expressed in breast cancer cells and that one isoform of Runx2 can activate bsp expression in these cells. In addition, we observe that bsp expression is additionally regulated by the homeodomain factor Msx2, another regulator of osteoblast-associated genes. Thus, this is the first report of osteoblast-related transcription factors being expressed in human breast cancer cells and provides a component of a mechanism that may explain the osteoblastic phenotype of human breast cancer cells that preferentially metastasize to bone. PMID- 12750291 TI - Evaluation of the prognostic potential of hyaluronic acid and hyaluronidase (HYAL1) for prostate cancer. AB - Despite the development of nomograms designed to evaluate the prognosis of a patient with prostate cancer (CaP), the information has been limited to prostate specific antigen (PSA), clinical stage, Gleason score, and tumor volume estimates. To improve our ability to predict prognosis, information regarding the molecular properties of CaP is needed. Hyaluronic acid (HA) is a glycosaminoglycan that promotes tumor metastasis. Hyaluronidase (HAase) is an enzyme that degrades HA into angiogenic fragments. We recently showed that in CaP tissues, whereas HA is localized mostly in the tumor-associated stroma, HYAL1 type HAase is exclusively localized in CaP cells (Lokeshwar et al. J. Biol. Chem., 276: 11922-11932, 2001). We evaluated the prognostic potential of HA and HYAL1 in CaP by immunohistochemistry. Archival CaP specimens were obtained from patients who underwent radical retropubic prostatectomy for clinically localized CaP. Group 1 (n = 25) included patients who showed biochemical recurrence (PSA >0.4 ng/ml; mean recurrence: 21.3 months). Group 2 included patients with no clinical or biochemical recurrence (n = 45; mean follow-up: 80.9 months). For HA and HYAL1 staining, a biotinylated HA-binding protein and an anti-HYAL1 antibody were used. The staining was evaluated on the basis of intensity (0 to 3+) and as dense or sparse (for HA staining only) and then grouped as low grade and high grade. In CaP specimens, HYAL1 was exclusively expressed in tumor cells. Although the stroma was stained positive for HA, 40% of tumor cells also expressed HA. HA, HYAL1, and combined HA-HYAL1 staining predicted progression with 96%, 84%, and 88% sensitivity, 55.5%, 80%, and 84.4% specificity, and 70%, 81.4%, and 85.7% accuracy, respectively. In the univariate analysis, preoperative PSA, Gleason sum, stage, margin, seminal vesicle, extra-prostatic extension (EPE), HA, HYAL1, and HA-HYAL1 were significant in predicting progression (P < 0.05). However, in the multiple logistic regression analysis, only EPE [odds ratio (OR) = 33.483; P = 0.002), HYAL1 (OR = 12.42; P = 0.009), HA-HYAL1 (OR = 18.048; P = 0.0033), and margin (OR = 26.948; P = 0.006)] were significant. Thus, in this 5-year follow-up study, HYAL1, together with EPE and margin, was found to be an independent prognostic indicator. PMID- 12750292 TI - Prostate-specific membrane antigen association with filamin A modulates its internalization and NAALADase activity. AB - Prostate-specific membrane antigen (PMSA) is an integral membrane protein highly expressed by prostate cancer cells. We reported previously that PSMA undergoes internalization via clathrin-coated pits (Liu et al., Cancer Res., 58: 4055-4060, 1998). In this study we demonstrate that filamin A, an actin cross-linking protein, associates with the cytoplasmic tail of PSMA and that this association of PSMA with filamin is involved in its localization to the recycling endosomal compartment. By ectopically expressing PSMA in filamin-negative and -positive cell lines, we additionally show that filamin binding to PSMA reduces the internalization rate of PSMA and its N-acelylated-alpha linked-acidic dipeptidase activity. These results suggest that filamin might be an important regulator of PSMA function. PMID- 12750293 TI - Molecular profiling of pancreatic adenocarcinoma and chronic pancreatitis identifies multiple genes differentially regulated in pancreatic cancer. AB - The molecular basis of pancreatic cancer is not understood. Previous attempts to determine the specific genes expressed in pancreatic cancer have been hampered by similarities between adenocarcinoma and chronic pancreatitis. In the current study, microarrays (Affymetrix) were used to profile gene expression in pancreatic adenocarcinoma (10), pancreatic cancer cell lines (7), chronic pancreatitis (5), and normal pancreas (5). Molecular profiling indicated a large number of genes differentially expressed between pancreatic cancer and normal pancreas but many fewer differences between pancreatic cancer and chronic pancreatitis, likely because of the shared stromal influences in the two diseases. To specifically identify genes expressed in neoplastic epithelium, we selected genes more highly expressed (>2-fold, p < 0.01) in adenocarcinoma compared with both normal pancreas and chronic pancreatitis and which were also highly expressed in pancreatic cancer cell lines. This strategy yielded 158 genes, of which 124 were not previously associated with pancreatic cancer. Quantitative-reverse transcription-PCR for two molecules, S100P and 14-3-3sigma, validated the microarray data. Support for the success of the neoplastic cell gene expression identification strategy was obtained by immunocytochemical localization of four representative genes, 14-3-3sigma, S100P, S100A6, and beta4 integrin, to neoplastic cells in pancreatic tumors. Thus, comparisons between pancreatic adenocarcinoma, pancreatic cancer cell lines, normal pancreas, and chronic pancreatitis have identified genes that are selectively expressed in the neoplastic epithelium of pancreatic adenocarcinoma. These data provide new insights into the molecular pathology of pancreatic cancer that may be useful for detection, diagnosis, and treatment. PMID- 12750294 TI - Transactivation of vimentin by beta-catenin in human breast cancer cells. AB - The cytoplasmic and nuclear redistribution of beta-catenin and the de novo expression of vimentin are frequently involved in the epithelial-to-mesenchymal transition associated with increased invasive/migratory properties of epithelial cells. Because beta-catenin can act as a coactivator of transcription through its binding to the T-cell factor (TCF)/lymphoid enhancer factor 1 transcription factor family, we have explored the possibility that beta-catenin/TCF could directly transactivate vimentin. We first compared vimentin expression in relation with the localization of beta-catenin in eight breast cancer cell lines displaying various degrees of invasiveness and in a model of cell migration using human mammary MCF10A cells. We could thus show a cytoplasmic and/or nuclear distribution of beta-catenin in invasive/migratory cells expressing vimentin, but not in noninvasive/stationary vimentin-negative cell lines. In addition, the human vimentin promoter was found to be up-regulated by beta-catenin and TCF-4 cotransfection. Varying with the cellular background, a diminution of this up regulation was observed when the putative beta-catenin/TCF binding site of the vimentin promoter was mutated. Our results therefore demonstrate that the vimentin promoter is a target of the beta-catenin/TCF pathway and strongly suggest an implication of this regulation in epithelial cell migration/invasion. PMID- 12750295 TI - EWI2/PGRL associates with the metastasis suppressor KAI1/CD82 and inhibits the migration of prostate cancer cells. AB - Cancer metastasis suppressor KAI1/CD82 belongs to the tetraspanin superfamily and inversely correlates with the metastatic potential of a variety of cancers. The mechanism of KAI1/CD82-mediated metastasis suppression remains unclear. In this study, we found a M(r) 68,00 cell-surface protein physically associated with KAI1/CD82 and named it KASP: a KAI1/CD82-associated surface protein. Distinctive from known KAI1/CD82 associations that usually occur in the context of "tetraspanin web," the KAI1/CD82-KASP association is likely to be direct because it is: (a) highly stoichiometric; (b) stabilized by chemical cross-linking; and (c) independent of cholesterol-enriched lipid rafts. Therefore, KASP is one of the major transmembrane proteins that associates with KAI1/CD82. Consistent with the wide distribution of KAI1/CD82, KASP is expressed ubiquitously in human tissues. Through peptide sequencing, KASP was identified as an immunoglobulin superfamily member called EWI2 or PGRL. Although EWI2/PGRL has been found to associate with tetraspanins CD9 and CD81, it forms distinct complexes with different tetraspanins, and its association with KAI1/CD82 could be independent of CD81 and CD9. Overexpression of EWI2/PGRL in Du145 metastatic prostate cancer cells inhibits cell migration on both fibronectin- and laminin-coated substratum, indicating that EWI2/PGRL directly regulates cell migration. Furthermore, EWI2/PGRL synergizes KAI1/CD82 in inhibiting cell migration, indicating that EWI2/PGRL is likely required for the function of KAI1/CD82. In summary, we identified a major KAI1/CD82-associated protein, EWI2/PGRL, that is important for KAI1/CD82-mediated suppression of cancer cell migration. PMID- 12750296 TI - Up-regulation of hypoxia-inducible factor 2alpha in renal cell carcinoma associated with loss of Tsc-2 tumor suppressor gene. AB - In the Eker rat model, inactivation of the Tuberous Sclerosis-2 (Tsc-2) tumor suppressor gene leads to high frequency of spontaneous renal cell carcinoma (RCC). By analogy to human RCC in which mutations in the von Hippel-Lindau (VHL) tumor suppressor gene result in accumulation of hypoxia-inducible factor alpha (HIFalpha) and up-regulation of vascular endothelial growth factor (VEGF), we investigated the regulation of HIF and its target gene VEGF in rat RCC resulting from Tsc-2 defects. To examine HIFalpha activity, a panel of rat renal epithelial cells were analyzed for expression of HIF1alpha and the homologous protein, HIF2alpha, under normoxic and hypoxic conditions. RCC-derived cell lines exhibited high basal levels of HIF activity as determined using hypoxia response element-luciferase reporter constructs. HIF2alpha was stabilized in RCC-derived cell lines and in five of six primary tumors compared with normal kidney, which was consistent with the high levels of hypoxia response element-reporter activity observed in the cell lines. Primary RCCs that developed in Eker rats were highly vascularized, which was similar to their human counterparts. Furthermore, reverse transcriptase PCR and immunoblotting demonstrated that VEGF was abundantly expressed in both rat RCC cell lines and primary tumors. The 120-, 164-, and 188 amino-acid isoforms of VEGF were expressed at the RNA and protein levels in RCC derived cell lines, although only a single band was observed in primary tumors. Taken together, these data suggest that RCC caused by loss of the Tsc-2 tumor suppressor gene (which retain wild-type Vhl) up-regulate VEGF via a HIF2alpha mediated mechanism. Thus, loss of Tsc-2 and VHL tumor suppressor gene function appears to have similar consequences in Eker rats and humans respectively, identifying dysregulation of HIFalpha and VEGF expression as a common pathway for the development of RCC in different species and in tumors with different molecular etiologies. PMID- 12750297 TI - 3'-[18F]fluoro-3'-deoxythymidine ([18F]-FLT) as positron emission tomography tracer for imaging proliferation in a murine B-Cell lymphoma model and in the human disease. AB - Here we describe the evaluation of 3'-[(18)F]fluoro-3'-deoxythymidine [[(18)F] FLT] as a tracer for positron emission tomography (PET) in a murine model of B cell lymphoma and in human malignant lymphoma. The human B-cell line DoHH2 expressed high levels of active thymidine kinase 1 (TK-1) as the key enzyme of [(18)F]-FLT metabolism. Immunostaining confirmed high levels of TK-1 in DoHH2 derived xenograft tumors in SCID/SCID mice. In vitro studies demonstrated a time dependent uptake of [(18)F]-FLT, an efficient phosphorylation to the respective monophosphate and the incorporation of [(18)F]-FLT into the perchloric acid insoluble fraction in DoHH2 cells, indicating the incorporation of this tracer into the DNA. After incubation with [(18)F]FLT for 240 min, 12.5% +/- 1.0% of radioactivity applied to the medium was intracellularly trapped in DoHH2 cells. Specific accumulation of [(18)F]-FLT in the malignant cell clone was confirmed in biodistribution studies in SCID/SCID mice bearing DoHH2-derived tumors. The percentage of injected dose of [(18)F]-FLT per gram of tumor tissue correlated with the tumor-proliferation index as evaluated in BrdUrd-labeling experiments. In a pilot study of 11 patients with both indolent and aggressive lymphoma, [(18)F]-FLT was suitable and comparable to [(18)F]-FDG in the ability to detect malignant lesions by PET scan. Furthermore, we found a close correlation (r = 0.95, P < 0.005) of the [(18)F]-FLT standardized uptake values with the Ki67 labeling index of tissue biopsies (n = 10) in these patients. These results suggest that [(18)F]-FLT represents a novel tracer for PET that enables imaging of proliferation in human lymphoma in vivo. PMID- 12750298 TI - Genetic reversion in an acute myelogenous leukemia cell line from a Fanconi anemia patient with biallelic mutations in BRCA2. AB - A 2-year old boy was diagnosed with Fanconi anemia (FA) and acute myeloid leukemia (AML). A cell line (termed FA-AML1) was established from blast cells obtained after a second relapse after a successful bone marrow transplant. Histochemical and surface marker analysis confirmed that the cells were derived from the myeloid lineage. Cytogenetic analysis revealed multiple chromosomal aberrations, including a ring 7. Stable proliferation of the cultured cells was absolutely dependent on the presence of granulocyte macrophage colony-stimulating factor or interleukin 3. This is the first AML cell line successfully established from a FA patient. Remarkably, FA-AML1 cells appeared to lack the characteristic cellular FA phenotype, i.e., a hypersensitivity to growth inhibition and chromosomal breakage by the cross-linking agent mitomycin C. Genomic DNA from the patient showed biallelic mutations [8415G>T (K2729N)and 8732C>A (S2835STOP)] in the breast cancer susceptibility gene FANCD1/BRCA2 [N. Howlett et al., Science (Wash. DC), 297: 606-609, 2002]. In the AML cells, however, the 8732C>A nonsense mutation was changed into a missense mutation by a secondary alteration, 8731T>G, resulting in 2835E, which restored the open-reading frame of the gene and could explain the reverted phenotype of these cells. Loss of the FA phenotype by genetic correction of a FA gene mutation during AML progression may be a common late event in the pathogenesis of AML in FA patients, which may be treatment related. This finding suggests a novel mechanistic principle of tumor progression based on the genetic correction of an early caretaker gene defect. PMID- 12750299 TI - Seroreactivity to epidermodysplasia verruciformis-related human papillomavirus types is associated with nonmelanoma skin cancer. AB - DNA from epidermodysplasia verruciformis-related human papillomavirus (EV-HPV) types is frequently found in nonmelanoma skin cancer (squamous and basal cell carcinoma). Epidemiological studies that investigate the relation between EV-HPV infection and nonmelanoma skin cancer are scarce. We designed a case-control study in which we looked for HPV infection in 540 cases with a history of skin cancer and 333 controls. By measuring seroreactivity to L1 virus-like particles of EV-HPV types 5, 8, 15, 20, 24, and 38 and the genital type HPV16 and by estimating the skin cancer relative risk among HPV seropositives, we analyzed whether EV-HPV serorecognition is associated with nonmelanoma skin cancer. Seroreactivity to five of the six EV-HPV types tested (HPV5, 8, 15, 20, and 24) was significantly increased in the squamous cell carcinoma cases. After adjusting for age and sex, the estimated squamous cell carcinoma relative risk was significantly increased in HPV8 and HPV38 seropositives [odds ratio (OR) = 14.7 (95% confidence interval (CI), 1.6-135) and OR = 3.0 (95% CI, 1.1-8.4), respectively]. The estimated relative risk for nodular and superficial multifocal basal cell carcinoma was also significantly increased in the HPV8 seropositives [OR = 9.2 (95% CI, 1.1-78.2) and OR = 17.3 (95% CI, 2.1-143), respectively] and in the HPV20 seropositives [OR = 3.2 (95% CI 1.3-7.9) and OR = 3.4 (95% CI 1.2 9.5), respectively]. The relative risk of developing malignant melanoma was not increased among HPV seropositives, and no associations were found for HPV16. Restricted analyses among the HPV seropositives only, to exclude distortion by interindividual differences in seroresponsiveness, underscored the significance of our findings. Restricted analyses among patients with skin cancer only, however, revealed that EV-HPV seropositivity was not significantly more present in patients with nonmelanoma skin cancer than in those with melanoma skin cancer. Taken together, our results indicate that EV-HPV serorecognition is nonspecifically associated with nonmelanoma skin cancer and suggest that EV-HPV directed seroresponses are induced upon skin cancer formation, rather than upon infection. PMID- 12750300 TI - Correspondence re: B-L. Adam et al., Serum protein fingerprinting coupled with a pattern-matching algorithm distinguishes prostate cancer from benign prostate hyperplasia and healthy men. Cancer Res., 62: 3609-3614, 2002. PMID- 12750303 TI - CD40-CD40L and platelet function: beyond hemostasis. PMID- 12750302 TI - Tachy- or bradyarrhythmias: implications for therapeutic intervention in LQT3 families. PMID- 12750304 TI - Angiopoietin-1 and pulmonary hypertension: cause or cure? PMID- 12750306 TI - Organelle proteomics: implications for subcellular fractionation in proteomics. AB - Functional proteome analysis is not restricted to the sequence information but includes the broad spectrum of structural modifications and quantitative changes of proteins to which they are subjected in different tissues and cell organelles and during the development of an organism. Cell biology has provided the means required for the analysis of the composition and properties of purified cellular elements. Subcellular fractionation is an approach universal across all cell types and tissues, including cardiac and vascular system. Subcellular fractionation and proteomics form an ideal partnership when it comes to enrichment and analysis of intracellular organelles and low abundant multiprotein complexes. Subcellular fractionation is a flexible and adjustable approach resulting in reduced sample complexity and is most efficiently combined with high resolution 2D gel/mass spectrometry analysis as well as with gel-independent techniques. In this study we introduce state of the art subcellular fractionation techniques and discuss their suitability, advantages, and limitations for proteomics research. PMID- 12750307 TI - Egr-1, a major link between infection and atherosclerosis? PMID- 12750305 TI - Genome informatics: current status and future prospects. AB - This article reviews recent advances in genomics and informatics relevant to cardiovascular research. In particular, we review the status of (1) whole genome sequencing efforts in human, mouse, rat, zebrafish, and dog; (2) the development of data mining and analysis tools; (3) the launching of the National Heart, Lung, and Blood Institute Programs for Genomics Applications and Proteomics Initiative; (4) efforts to characterize the cardiac transcriptome and proteome; and (5) the current status of computational modeling of the cardiac myocyte. In each instance, we provide links to relevant sources of information on the World Wide Web and critical appraisals of the promises and the challenges of an expanding and diverse information landscape. PMID- 12750308 TI - Genetically determined heterogeneity in hemoglobin scavenging and susceptibility to diabetic cardiovascular disease. AB - A major function of haptoglobin (Hp) is to bind hemoglobin (Hb) to form a stable Hp-Hb complex and thereby prevent Hb-induced oxidative tissue damage. Clearance of the Hp-Hb complex can be mediated by the monocyte/macrophage scavenger receptor CD163. We recently demonstrated that diabetic individuals homozygous for the Hp 2 allele (Hp 2-2) were at 500% greater risk of cardiovascular disease (CVD) compared with diabetic individuals homozygous for the Hp 1 allele (Hp 1-1). No differences in risk by Hp type were seen in individuals without diabetes. To understand the relationship between the Hp polymorphism and diabetic CVD, we sought to identify differences in antioxidant and scavenging functions between the Hp types and to determine how these functions were modified in diabetes. The scavenging function of Hp was assessed using rhodamine-tagged and 125I-Hp in cell lines stably transfected with CD163 and in macrophages expressing endogenous CD163. We found that the rate of clearance of Hp 1-1-Hb by CD163 is markedly greater than that of Hp 2-2-Hb. Diabetes is associated with an increase in the nonenzymatic glycosylation of serum proteins, including Hb. The antioxidant function of Hp was assessed with glycosylated and nonglycosylated Hb. We identified a severe impairment in the ability of Hp to prevent oxidation mediated by glycosylated Hb. We propose that the specific interaction between diabetes, CVD, and Hp genotype is the result of the heightened urgency of rapidly clearing glycosylated Hb-Hp complexes from the subendothelial space before they can oxidatively modify low-density lipoprotein to atherogenic oxidized low-density lipoprotein. PMID- 12750310 TI - Matrix metalloproteinase-3 genotype contributes to age-related aortic stiffening through modulation of gene and protein expression. AB - Matrix metalloproteinases (MMPs) include most major constituents of the arterial wall as substrates. A common promotor polymorphism (5A/6A) is associated with differences in MMP-3 (stromelysin-1) activity, and associations with certain forms of vascular disease have been shown. This study investigated whether the MMP-3 5A/6A promoter polymorphism contributes to age-related large artery stiffening. MMP-3 5A/6A genotype was determined in 203 (135 male) low cardiovascular risk, unmedicated individuals who were divided prospectively into two groups (30 to 60 years, n=126; > or =61 years, n=77). Noninvasive large artery stiffness was measured as ascending aortic input impedance from brachial blood pressure, carotid tonometry, and Doppler ascending aortic blood flow. In the older group, homozygotes had higher aortic input (P<0.01) and characteristic (P<0.01) impedance, ie, higher stiffness, than heterozygotes after correction for the effects of age, gender, and mean arterial pressure. There was no such difference in the younger group. Gene expression was subsequently investigated in dermal biopsies in randomly selected older men from the same cohort with real time PCR (n=40). In 5A homozygotes, gene expression was 4-fold higher (P<0.05), and in 6A homozygotes, 2-fold lower (P<0.05) compared with the heterozygotes. Differences in gene expression were associated with corresponding significant changes in MMP-3 protein levels. Concordance between dermal and aortic gene and protein expression was shown in a separate cohort of postmortem aortic samples (n=7). We conclude that MMP-3 genotype may be an important determinant of vascular remodeling and age-related arterial stiffening, with the heterozygote having the optimal balance between matrix accumulation and deposition. PMID- 12750309 TI - HFE mutation and dietary iron content interact to increase ischemia/reperfusion injury of the heart in mice. AB - Hereditary hemochromatosis is an inherited pathological condition characterized by iron overload in several vital organs including heart. To increase our understanding of the underlying pathogenic mechanisms of hereditary hemochromatosis, we used a HFE gene knockout mouse model that replicates hereditary hemochromatosis. A group of mice with no copies of HFE gene and corresponding wild-type mice were maintained either on low-iron (30 ppm) or high iron (300 ppm) diet since birth. The results of our study revealed that HFE gene knockout mouse hearts were susceptible to ischemia-reperfusion injury as evidenced by increased postischemic ventricular dysfunction, increased myocardial infarct size and cardiomyocyte apoptosis compared with wild-type control hearts. The degree of injury increased in the hearts of the mice fed high-iron diet. The hearts of the HFE knockout mice showed increased iron deposition, increased content of reactive oxygen species (ROS) as evidenced by the increased formation of malondialdehyde, and reduced antioxidant enzymes including superoxide dismutase, catalase, and glutathione peroxidase. The results suggest that increased amount of ROS and reduced antioxidant reserve secondary to iron overloading may be instrumental for the susceptibility of the HFE gene knockout mice to cardiac injury. PMID- 12750311 TI - Persistent stunning induces myocardial hibernation and protection: flow/function and metabolic mechanisms. AB - To test the hypothesis that persistent myocardial stunning can lead to hibernating myocardium, 13 pigs were chronically instrumented, and persistent stunning was induced regionally by 6 repetitive episodes of 90-minute coronary stenosis (CS) (30% reduction in baseline coronary blood flow [CBF]) followed by full reperfusion every 12 hours. During the 1st CS, CBF fell from 43+/-2 to 31+/ 2 mL/min, and anterior wall thickening (AWT) fell by 54+/-8%, but posterior WT did not change. AWT never recovered fully and remained depressed by 31+/-7% before the 6th CS, reflecting persistent myocardial stunning, but baseline CBF was not changed. Surprisingly, during the 6th CS, AWT did not fall further despite a similar reduction in CBF during CS, as occurred with the 1st episode. Regional Mo2 fell similarly during the 1st and 6th CS. During the 1st CS, plasma glucose uptake increased, whereas free fatty acid (FFA) uptake was reduced. Before the 6th CS, glucose uptake remained elevated, whereas FFA uptake remained reduced. Histology revealed enhanced glycogen deposition, which could be explained by decreased glycogen synthase kinase (GSK)-3beta protein levels and activity. These results indicate that persistent stunning, even in the absence of chronic ischemia, can recapitulate the phenotype of myocardial hibernation. This results in a shift in the flow/function relationship where a 30% decrease in CBF is no longer accompanied by a fall in myocardial function, which could be explained, in part, by a shift in substrate utilization. These hemodynamic/metabolic adjustments could facilitate survival of hibernating myocardium. PMID- 12750312 TI - Modification of GATA-2 transcriptional activity in endothelial cells by the SUMO E3 ligase PIASy. AB - GATA sequences are required for the optimal expression of endothelial cell specific genes, including endothelin-1 (ET-1). We have identified PIASy in a search for new GATA-2 interacting proteins that can regulate GATA-2-mediated endothelial gene expression. Notably, among the cell populations comprising vascular walls, PIASy mRNA is selectively expressed in endothelial cells, and its expression can be regulated by angiogenic growth factors. We show that GATA-2 is covalently modified by small ubiquitin-like modifier (SUMO)-1 and -2 and that PIASy, through its E3 SUMO ligase activity, preferentially enhances the conjugation of SUMO-2 to GATA-2. Through a functional analysis, we demonstrate that PIASy potently suppresses the activity of the GATA-2-dependent human ET-1 promoter in endothelial cells. The suppressive effect of PIASy requires the GATA binding site in the ET-1 promoter and depends on its interaction with GATA-2, which requires both N-terminal (amino acids 1-183) and C-terminal (amino acids 414-510) sequences in PIASy. We conclude that PIASy enhances the conjugation of SUMO-2 to GATA-2 and that the interaction of PIASy with GATA-2 can modulate GATA mediated ET-1 transcription activity in endothelial cells through a RING-like domain-independent mechanism. PMID- 12750313 TI - Mechanical stretch enhances mRNA expression and proenzyme release of matrix metalloproteinase-2 (MMP-2) via NAD(P)H oxidase-derived reactive oxygen species. AB - Mechanical stretch is a hallmark of arterial hypertension and leads to vessel wall remodeling, which involves matrix metalloproteinases (MMPs). Because mechanical stretch is further capable of inducing reactive oxygen species (ROS) formation via the NAD(P)H oxidase, we assessed whether mechanical stretch enhances MMP expression and activity in a NAD(P)H oxidase-dependent manner. Therefore, vascular smooth muscle cells (VSMCs) isolated from C57BL/6 mice were exposed to cyclic mechanical stretch. The impact of ROS was assessed using VSMCs isolated from p47phox-/- mice, deficient for a NAD(P)H oxidase subunit responsible for ROS formation. Transcript levels were investigated by cDNA array and confirmed by RT-PCR. ROS formation was determined by DCF fluoroscopy and MMP 2 activity by zymography. Mechanical stretch of wild-type VSMCs resulted in a rapid ROS formation and p47phox membrane translocation that is followed by an increase in Nox-1 transcripts. ROS formation was completely abrogated in p47phox /- VSMCs. cDNA array further revealed an increase of MMP-2 mRNA in response to mechanical stretch, which was validated by RT-PCR. Using p47phox-/- VSMCs, this increase in MMP-2 mRNA was completely blunted. mRNA expression of tissue inhibitor of MMP-2 TIMP-1 and TIMP-2 and membrane-type 1 MMP was unaffected by mechanical stretch. Gelatinolytic activity of pro-MMP-2 has been increased rapidly in wild-type VSMCs and was completely abolished in p47phox-/- VSMCs. These results indicate that mechanical stretch induces ROS formation via the NAD(P)H oxidase and thereby enhances MMP-2 mRNA expression and pro-MMP-2 release. These results are consistent with the notion that in arterial hypertension, reactive oxygen species are involved in vascular remodeling via MMP activation. The full text of this article is available online at http://www.circresaha.org. PMID- 12750314 TI - NFATc3 and NFATc4 are required for cardiac development and mitochondrial function. AB - Activation of the nuclear factor of activated T-cell (NFAT) family of transcription factors is associated with changes in gene expression and myocyte function in adult cardiac and skeletal muscle. However, the role of NFATs in normal embryonic heart development is not well characterized. In this report, the function of NFATc3 and NFATc4 in embryonic heart development was examined in mice with targeted disruption of both nfatc3 and nfatc4 genes. The nfatc3-/-nfatc4-/- mice demonstrate embryonic lethality after embryonic day 10.5 and have thin ventricles, pericardial effusion, and a reduction in ventricular myocyte proliferation. Cardiac mitochondria are swollen with abnormal cristae, indicative of metabolic failure, but hallmarks of apoptosis are not evident. Furthermore, enzymatic activity of complex II and IV of the respiratory chain and mitochondrial oxidative activity are reduced in nfatc3-/-nfatc4-/- cardiomyocytes. Cardiac-specific expression of constitutively active NFATc4 in nfatc3-/-nfatc4-/- embryos prolongs embryonic viability to embryonic day 12 and preserves ventricular myocyte proliferation, compact zone density, and trabecular formation. The rescued embryos also maintain cardiac mitochondrial ultrastructure and complex II enzyme activity. Together, these data support the hypothesis that loss of NFAT activity in the heart results in a deficiency in mitochondrial energy metabolism required for cardiac morphogenesis and function. PMID- 12750315 TI - C. C. Tan: a life of peaks and valleys. PMID- 12750316 TI - Male-killing Wolbachia and mitochondrial DNA: selective sweeps, hybrid introgression and parasite population dynamics. AB - Mitochondrial DNA (mtDNA) sequences are widely used as neutral genetic markers in insects. However, patterns of mtDNA variability are confounded by the spread of maternally transmitted parasites, which are genetically linked to the mitochondria. We have investigated these effects in the butterflies Acraea encedon (which is host to two strains of male-killing Wolbachia bacteria) and A. encedana (which is host to one strain). Within a population, the mitochondria are in linkage disequilibrium with the different male-killers. Furthermore, there has been a recent selective sweep of the mtDNA, which has led to the loss of mitochondrial variation within populations and erased any geographical structure. We also found that one of the male-killers, together with the associated mtDNA, has introgressed from A. encedana into A. encedon within the last 16,000 years. Interestingly, because butterflies are female heterogametic, this will presumably have also led to the introgression of genes on the W sex chromosome. Finally, in A. encedon the mitochondria in uninfected females are unaltered by the spread of the male-killer and have diverse, geographically structured mtDNA. This means we can reject the hypothesis that the male-killer is at a stable equilibrium maintained by imperfect transmission of the bacterium. Instead, some other form of balancing selection may be maintaining uninfected females in the population and preventing the species from going extinct due to a shortage of males. PMID- 12750318 TI - Experimental prediction of the evolution of cefepime resistance from the CMY-2 AmpC beta-lactamase. AB - Understanding of the evolutionary histories of many genes has not yet allowed us to predict the evolutionary potential of those genes. Intuition suggests that current biochemical activity of gene products should be a good predictor of the potential to evolve related activities; however, we have little evidence to support that intuition. Here we use our in vitro evolution method to evaluate biochemical activity as a predictor of future evolutionary potential. Neither the class C Citrobacter freundii CMY-2 AmpC beta-lactamase nor the class A TEM-1 beta lactamase confer resistance to the beta-lactam antibiotic cefepime, nor do any of the naturally occurring alleles descended from them. However, the CMY-2 AmpC enzyme and some alleles descended from TEM-1 confer high-level resistance to the structurally similar ceftazidime. On the basis of the comparison of TEM-1 and CMY 2, we asked whether biochemical activity is a good predictor of the evolutionary potential of an enzyme. If it is, then CMY-2 should be more able than the TEMs to evolve the ability to confer higher levels of cefepime resistance. Although we generated CMY-2 evolvants that conferred increased cefepime resistance, we did not recover any CMY-2 evolvants that conferred resistance levels as high as the best cefepime-resistant TEM alleles. PMID- 12750317 TI - Horizontal acquisition of divergent chromosomal DNA in bacteria: effects of mutator phenotypes. AB - We examine the potential beneficial effects of the expanded access to environmental DNA offered by mutators on the adaptive potential of bacterial populations. Using parameters from published studies of recombination in E. coli, we find that the presence of mutators has the potential to greatly enhance bacterial population adaptation when compared to populations without mutators. In one specific example, for which three specific amino acid substitutions are required for adaptation to occur in a 300-amino-acid protein, we found a 3500 fold increase in the rate of adaptation. The probability of a beneficial acquisition decreased if more amino acid changes, or integration of longer DNA fragments, were required for adaptation. The model also predicts that mutators are more likely than nonmutator phenotypes to acquire genetic variability from a more diverged set of donor bacteria. Bacterial populations harboring mutators in a sequence heterogeneous environment are predicted to acquire most of their DNA conferring adaptation in the range of 13-30% divergence, whereas nonmutator phenotypes become adapted after recombining with more homogeneous sequences of 7 21% divergence. We conclude that mutators can accelerate bacterial adaptation when desired genetic variability is present within DNA fragments of up to approximately 30% divergence. PMID- 12750319 TI - Positive and negative roles of homologous recombination in the maintenance of genome stability in Saccharomyces cerevisiae. AB - In previous studies of the loss of heterozygosity (LOH), we analyzed a hemizygous URA3 marker on chromosome III in S. cerevisiae and showed that homologous recombination is involved in processes that lead to LOH in multiple ways, including allelic recombination, chromosome size alterations, and chromosome loss. To investigate the role of homologous recombination more precisely, we examined LOH events in rad50 Delta, rad51 Delta, rad52 Delta, rad50 Delta rad52 Delta, and rad51 Delta rad52 Delta mutants. As compared to Rad(+) cells, the frequency of LOH was significantly increased in all mutants, and most events were chromosome loss. Other LOH events were differentially affected in each mutant: the frequencies of all types of recombination were decreased in rad52 mutants and enhanced in rad50 mutants. The rad51 mutation increased the frequency of ectopic but not allelic recombination. Both the rad52 and rad51 mutations increased the frequency of intragenic point mutations approximately 25-fold, suggesting that alternative mutagenic pathways partially substitute for homologous recombination. Overall, these results indicate that all of the genes are required for chromosome maintenance and that they most likely function in homologous recombination between sister chromatids. In contrast, other recombination pathways can occur at a substantial level even in the absence of one of the genes and contribute to generating various chromosome rearrangements. PMID- 12750320 TI - Multiple roles for Saccharomyces cerevisiae histone H2A in telomere position effect, Spt phenotypes and double-strand-break repair. AB - Telomere position effects on transcription (TPE, or telomeric silencing) are nucleated by association of nonhistone silencing factors with the telomere and propagated in subtelomeric regions through association of silencing factors with the specifically modified histones H3 and H4. However, the function of histone H2A in TPE is unknown. We found that deletion of either the amino or the carboxyltails of H2A substantially reduces TPE. We identified four H2A modification sites necessary for wild-type efficiency of TPE. These "hta1tpe" alleles also act as suppressors of a delta insertion allele of LYS2, suggesting shared elements of chromatin structure at both loci. Interestingly, we observed combinatorial effects of allele pairs, suggesting both interdependent acetylation and deacetylation events in the amino-terminal tail and a regulatory circuit between multiple phosphorylated residues in the carboxyl-terminal tail. Decreases in silencing and viability are observed in most hta1tpe alleles after treatment with low and high concentrations, respectively, of bleomycin, which forms double strand breaks (DSBs). In the absence of the DSB and telomere-binding protein yKu70, the bleomycin sensitivity of hta1tpe alleles is further enhanced. We also provide data suggesting the presence of a yKu-dependent histone H2A function in TPE. These data indicate that the amino- and carboxyl-terminal tails of H2A are essential for wild-type levels of yKu-mediated TPE and DSB repair. PMID- 12750321 TI - Characterization of the hyperrecombination phenotype of the pol3-t mutation of Saccharomyces cerevisiae. AB - The DNA polymerase delta (Pol3p/Cdc2p) allele pol3-t of Saccharomyces cerevisiae has previously been shown to increase the frequency of deletions between short repeats (several base pairs), between homologous DNA sequences separated by long inverted repeats, and between distant short repeats, increasing the frequency of genomic deletions. We found that the pol3-t mutation increased intrachromosomal recombination events between direct DNA repeats up to 36-fold and interchromosomal recombination 14-fold. The hyperrecombination phenotype of pol3 t was partially dependent on the Rad52p function but much more so on Rad1p. However, in the double-mutant rad1 Delta rad52 Delta, the pol3-t mutation still increased spontaneous intrachromosomal recombination frequencies, suggesting that a Rad1p Rad52p-independent single-strand annealing pathway is involved. UV and gamma-rays were less potent inducers of recombination in the pol3-t mutant, indicating that Pol3p is partly involved in DNA-damage-induced recombination. In contrast, while UV- and gamma-ray-induced intrachromosomal recombination was almost completely abolished in the rad52 or the rad1 rad52 mutant, there was still good induction in those mutants in the pol3-t background, indicating channeling of lesions into the above-mentioned Rad1p Rad52p-independent pathway. Finally, a heterozygous pol3-t/POL3 mutant also showed an increased frequency of deletions and MMS sensitivity at the restrictive temperature, indicating that even a heterozygous polymerase delta mutation might increase the frequency of genetic instability. PMID- 12750322 TI - The Mus81/Mms4 endonuclease acts independently of double-Holliday junction resolution to promote a distinct subset of crossovers during meiosis in budding yeast. AB - Current models for meiotic recombination require that crossovers derive from the resolution of a double-Holliday junction (dHJ) intermediate. In prokaryotes, enzymes responsible for HJ resolution are well characterized but the identification of a eukaryotic nuclear HJ resolvase has been elusive. Indirect evidence suggests that MUS81 from humans and fission yeast encodes a HJ resolvase. We provide three lines of evidence that Mus81/Mms4 is not the major meiotic HJ resolvase in S. cerevisiae: (1) MUS81/MMS4 is required to form only a distinct subset of crossovers; (2) rather than accumulating, dHJ intermediates are reduced in an mms4 mutant; and (3) expression of a bacterial HJ resolvase has no suppressive effect on mus81 meiotic phenotypes. Our analysis also reveals the existence of two distinct classes of crossovers in budding yeast. Class I is dependent upon MSH4/MSH5 and exhibits crossover interference, while class II is dependent upon MUS81/MMS4 and exhibits no interference. mms4 specifically reduces crossing over on small chromosomes, which are known to undergo less interference. The correlation between recombination rate and degree of interference to chromosome size may therefore be achieved by modulating the balance between class I/class II crossovers. PMID- 12750323 TI - Molecular characterization and analysis of the acrB gene of Aspergillus nidulans: a gene identified by genetic interaction as a component of the regulatory network that includes the CreB deubiquitination enzyme. AB - Mutations in the acrB gene, which were originally selected through their resistance to acriflavine, also result in reduced growth on a range of sole carbon sources, including fructose, cellobiose, raffinose, and starch, and reduced utilization of omega-amino acids, including GABA and beta-alanine, as sole carbon and nitrogen sources. The acrB2 mutation suppresses the phenotypic effects of mutations in the creB gene that encodes a regulatory deubiquitinating enzyme, and in the creC gene that encodes a WD40-repeat-containing protein. Thus AcrB interacts with a regulatory network controlling carbon source utilization that involves ubiquitination and deubiquitination. The acrB gene was cloned and physically analyzed, and it encodes a novel protein that contains three putative transmembrane domains and a coiled-coil region. AcrB may play a role in the ubiquitination aspect of this regulatory network. PMID- 12750324 TI - Theory of time-resolved somatic complementation and its use to explore the sporulation control network in Physarum polycephalum. AB - Mutants of Physarum polycephalum can be complemented by fusion of plasmodial cells followed by cytoplasmic mixing. Complementation between strains carrying different mutational defects in the sporulation control network may depend on the signaling state of the network components. We have previously suggested that time resolved somatic complementation (TRSC) analysis with such mutants may be used to probe network architecture and dynamics. By computer simulation it is now shown how and under which conditions the regulatory hierarchy of genes can be determined experimentally. A kinetic model of the sporulation control network is developed, which is then used to demonstrate how the mechanisms of TRSC can be understood and simulated at the kinetic level. On the basis of theoretical considerations, experimental parameters that determine whether functional complementation of two mutations will occur are identified. It is also shown how gene dosage-effect relationships can be employed for network analysis. The theoretical framework provided may be used to systematically analyze network structure and dynamics through time-resolved somatic complementation studies. The conclusions drawn are of general relevance in that they do not depend on the validity of the model from which they were derived. PMID- 12750325 TI - Single-nucleotide polymorphisms of the Trypanosoma cruzi MSH2 gene support the existence of three phylogenetic lineages presenting differences in mismatch repair efficiency. AB - We have identified single-nucleotide polymorphisms (SNPs) in the mismatch-repair gene TcMSH2 from Trypanosoma cruzi. Phylogenetic inferences based on the SNPs, confirmed by RFLP analysis of 32 strains, showed three distinct haplogroups, denominated A, B, and C. Haplogroups A and C presented strong identity with the previously described T. cruzi lineages I and II, respectively. A third haplogroup (B) was composed of strains presenting hybrid characteristics. All strains from a haplogroup encoded the same specific protein isoform, called, respectively, TcMHS2a, TcMHS2b, and TcMHS2c. The classification into haplogroups A, B, and C correlated with variation in the efficiency of mismatch repair in these cells. When microsatellite loci of strains representative of each haplogroup were analyzed after being cultured in the presence of hydrogen peroxide, new microsatellite alleles were definitely seen in haplogroups B and C, while no evidence of microsatellite instability was found in haplogroup A. Also, cells from haplogroups B and C were considerably more resistant to cisplatin treatment, a characteristic known to be conferred by deficiency of mismatch repair in eukaryotic cells. Altogether, our data suggest that strains belonging to haplogroups B and C may have decreased mismatch-repair ability when compared with strains assigned to the haplogroup A lineage. PMID- 12750326 TI - Continuous exchange of sequence information between dispersed Tc1 transposons in the Caenorhabditis elegans genome. AB - In a genome-wide analysis of the active transposons in Caenorhabditis elegans we determined the localization and sequence of all copies of each of the six active transposon families. Most copies of the most active transposons, Tc1 and Tc3, are intact but individually have a unique sequence, because of unique patterns of single-nucleotide polymorphisms. The sequence of each of the 32 Tc1 elements is invariant in the C. elegans strain N2, which has no germline transposition. However, at the same 32 Tc1 loci in strains with germline transposition, Tc1 elements can acquire the sequence of Tc1 elements elsewhere in the N2 genome or a chimeric sequence derived from two dispersed Tc1 elements. We hypothesize that during double-strand-break repair after Tc1 excision, the template for repair can switch from the Tc1 element on the sister chromatid or homologous chromosome to a Tc1 copy elsewhere in the genome. Thus, the population of active transposable elements in C. elegans is highly dynamic because of a continuous exchange of sequence information between individual copies, potentially allowing a higher evolution rate than that found in endogenous genes. PMID- 12750328 TI - Mechanosensory inputs influence Caenorhabditis elegans pharyngeal activity via ivermectin sensitivity genes. AB - Mechanical stimulation induces opposite behavioral responses in the adult and dauer pharynx. Tail tap of adults inhibits pharyngeal pumping via a pathway involving the innexin gene unc-7 and components of the glutamatergic pathway encoded by the genes avr-14 and avr-15. Tail tap of dauers stimulates pumping through a mechanism involving G alpha o and G alpha q. The nematocidal drug ivermectin is believed to kill worms by opening a glutamate-gated chloride channel (AVR-15) on pharyngeal muscle, causing complete pumping inhibition. However, ivermectin can also inhibit pumping in the absence of this channel. We propose that one of the ways ivermectin could prevent pumping, in the absence of the AVR-15 ivermectin-binding channel on pharynx muscle, is to target AVR-14 and AVR-15, which are expressed in the inhibitory pathway linking mechanosensation and pumping activity. PMID- 12750327 TI - New genes that interact with lin-35 Rb to negatively regulate the let-60 ras pathway in Caenorhabditis elegans. AB - Previous studies have shown that a synthetic multivulva phenotype results from mutations in genes that antagonize the ras-mediated intercellular signaling system responsible for vulval induction in Caenorhabditis elegans. Synthetic multivulva mutations define two classes of genes, A and B, and a mutation in a gene of each class is required to produce the multivulva phenotype. The ectopic vulval tissue in multivulva animals is generated by vulval precursor cells that in the wild type do not generate vulval tissue. One of the class B synthetic multivulva genes, lin-35, encodes a protein similar to the retinoblastoma (Rb) protein. In this article, we describe the isolation and characterization of 50 synthetic multivulva mutations, the identification of new components of both the class A and class B lin-35 Rb pathways, and the cloning of lin-52, a class B gene that may have a conserved role in Rb-mediated signaling. PMID- 12750329 TI - Genetic analysis of a synaptic calcium channel in Drosophila: intragenic modifiers of a temperature-sensitive paralytic mutant of cacophony. AB - Our previous genetic analysis of synaptic mechanisms in Drosophila identified a temperature-sensitive paralytic mutant of the voltage-gated calcium channel alpha1 subunit gene, cacophony (cac). Electrophysiological studies in this mutant, designated cac(TS2), indicated cac encodes a primary calcium channel alpha1 subunit functioning in neurotransmitter release. To further examine the functions and interactions of cac-encoded calcium channels, a genetic screen was performed to isolate new mutations that modify the cac(TS2) paralytic phenotype. The screen recovered 10 mutations that enhance or suppress cac(TS2), including second-site mutations in cac (intragenic modifiers) as well as mutations mapping to other genes (extragenic modifiers). Here we report molecular characterization of three intragenic modifiers and examine the consequences of these mutations for temperature-sensitive behavior, synaptic function, and processing of cac pre mRNAs. These mutations may further define the structural basis of calcium channel alpha1 subunit function in neurotransmitter release. PMID- 12750331 TI - Mitochondrial genotype affects fitness in Drosophila simulans. AB - Drosophila simulans is known to harbor three distinct mitochondrial DNA (mtDNA) haplotype groups (siI, -II, and -III) with nearly 3.0% interhaplotypic divergence but <0.06% intrahaplotypic diversity. With the large amount of genetic variation in this system, the potential power to detect intraspecific fitness differences in fly lines that carry distinct haplotypes is great. We test three life-history traits on fly lines with known sequence differences in the mtDNA genome after controlling the nuclear genome by backcrossing. We find that flies with the siI haplotype are fastest developing and have the lowest probability of surviving to three experimental periods (2-6, 12-17, and 34-39 days of age). Wild-type males with siIII mtDNA were more active while disruption of specific coadapted nucleo mitochondrial complexes caused a significant decrease in activity. These results are discussed in the context of the geographic distribution of each haplotype. PMID- 12750330 TI - Logjam encodes a predicted EMP24/GP25 protein that is required for Drosophila oviposition behavior. AB - A newly characterized Drosophila melanogaster gene, logjam (loj), functions in female reproduction by modulating oviposition behavior. The locus encodes at least six overlapping transcripts with unique 5' ends. P-element mutants that express very low levels of loj transcripts are unable to oviposit mature eggs. This phenotype can be rescued by the introduction of a transgene expressing the most abundant loj transcript. As for many genes that specify behavioral outputs, loj is present in the adult central nervous system (CNS). Interestingly, it is also observed in vitellogenic egg chambers, suggesting that there may be multiple functions for this gene in egg-laying behavior. loj encodes a predicted protein with homology to the EMP24/GP25 transmembrane components of cytoplasmic vesicles and likely functions in intracellular trafficking. PMID- 12750332 TI - Telomeric associated sequences of Drosophila recruit polycomb-group proteins in vivo and can induce pairing-sensitive repression. AB - In Drosophila, relocation of a euchromatic gene near centromeric or telomeric heterochromatin often leads to its mosaic silencing. Nevertheless, modifiers of centromeric silencing do not affect telomeric silencing, suggesting that each location requires specific factors. Previous studies suggest that a subset of Polycomb-group (PcG) proteins could be responsible for telomeric silencing. Here, we present the effect on telomeric silencing of 50 mutant alleles of the PcG genes and of their counteracting trithorax-group genes. Several combinations of two mutated PcG genes impair telomeric silencing synergistically, revealing that some of these genes are required for telomeric silencing. In situ hybridization and immunostaining experiments on polytene chromosomes revealed a strict correlation between the presence of PcG proteins and that of heterochromatic telomeric associated sequences (TASs), suggesting that TASs and PcG complexes could be associated at telomeres. Furthermore, lines harboring a transgene containing an X-linked TAS subunit and the mini-white reporter gene can exhibit pairing-sensitive repression of the white gene in an orientation-dependent manner. Finally, an additional binding site for PcG proteins was detected at the insertion site of this type of transgene. Taken together, these results demonstrate that PcG proteins bind TASs in vivo and may be major players in Drosophila telomeric position effect (TPE). PMID- 12750333 TI - Suppression of muscle hypercontraction by mutations in the myosin heavy chain gene of Drosophila melanogaster. AB - The indirect flight muscles (IFM) of Drosophila melanogaster provide a good genetic system with which to investigate muscle function. Flight muscle contraction is regulated by both stretch and Ca(2+)-induced thin filament (actin + tropomyosin + troponin complex) activation. Some mutants in troponin-I (TnI) and troponin-T (TnT) genes cause a "hypercontraction" muscle phenotype, suggesting that this condition arises from defects in Ca(2+) regulation and actomyosin-generated tension. We have tested the hypothesis that missense mutations of the myosin heavy chain gene, Mhc, which suppress the hypercontraction of the TnI mutant held-up(2) (hdp(2)), do so by reducing actomyosin force production. Here we show that a "headless" Mhc transgenic fly construct that reduces the myosin head concentration in the muscle thick filaments acts as a dose-dependent suppressor of hypercontracting alleles of TnI, TnT, Mhc, and flightin genes. The data suggest that most, if not all, mutants causing hypercontraction require actomyosin-produced forces to do so. Whether all Mhc suppressors act simply by reducing the force production of the thick filament is discussed with respect to current models of myosin function and thin filament activation by the binding of calcium to the troponin complex. PMID- 12750334 TI - Host genotype determines cytoplasmic incompatibility type in the haplodiploid genus Nasonia. AB - In haplodiploid species, Wolbachia-induced cytoplasmic incompatibility (CI) can be expressed in one of two ways: as a "conversion" of diploid fertilized eggs into haploid males or as embryonic mortality. Here we describe CI-type variation within the parasitic wasp genus Nasonia and genetically analyze the basis of this variation. We reach four main conclusions: (i) CI is expressed primarily as conversion in N. vitripennis, but as embryonic mortality in the sibling species N. giraulti and N. longicornis; (ii) the difference in CI type between N. giraulti (mortality) and N. vitripennis (conversion) is determined by host nuclear genotype rather than by Wolbachia differences; (iii) N. vitripennis "conversion genes" are recessive in hybrid females; and (iv) a difference in CI level between the sibling species N. giraulti and N. longicornis is due to the different Wolbachia infections in the species rather than to the host genotype. These results show that host nuclear genes can influence the type of CI present in a species. On the basis of these findings, we propose a model for how different CI types evolve in haplodiploids due to selection on nuclear genes modifying CI. PMID- 12750336 TI - Drosophila gain-of-function mutant RTK torso triggers ectopic Dpp and STAT signaling. AB - Overactivation of receptor tyrosine kinases (RTKs) has been linked to tumorigenesis. To understand how a hyperactivated RTK functions differently from wild-type RTK, we conducted a genome-wide systematic survey for genes that are required for signaling by a gain-of-function mutant Drosophila RTK Torso (Tor). We screened chromosomal deficiencies for suppression of a gain-of-function mutation tor (tor(GOF)), which led to the identification of 26 genomic regions that, when in half dosage, suppressed the defects caused by tor(GOF). Testing of candidate genes in these regions revealed many genes known to be involved in Tor signaling (such as those encoding the Ras-MAPK cassette, adaptor and structural molecules of RTK signaling, and downstream target genes of Tor), confirming the specificity of this genetic screen. Importantly, this screen also identified components of the TGFbeta (Dpp) and JAK/STAT pathways as being required for Tor(GOF) signaling. Specifically, we found that reducing the dosage of thickveins (tkv), Mothers against dpp (Mad), or STAT92E (aka marelle), respectively, suppressed tor(GOF) phenotypes. Furthermore, we demonstrate that in tor(GOF) embryos, dpp is ectopically expressed and thus may contribute to the patterning defects. These results demonstrate an essential requirement of noncanonical signaling pathways for a persistently activated RTK to cause pathological defects in an organism. PMID- 12750335 TI - Ectopic expression of inhibitors of protein phosphatase type 1 (PP1) can be used to analyze roles of PP1 in Drosophila development. AB - We have identified two proteins that bind with high specificity to type 1 serine/threonine protein phosphatase (PP1) and have exploited their inhibitory properties to develop an efficient and flexible strategy for conditional inactivation of PP1 in vivo. We show that modest overexpression of Drosophila homologs of I-2 and NIPP1 (I-2Dm and NIPP1Dm) reduces the level of PP1 activity and phenotypically resembles known PP1 mutants. These phenotypes, which include lethality, abnormal mitotic figures, and defects in muscle development, are suppressed by coexpression of PP1, indicating that the effect is due specifically to loss of PP1 activity. Reactivation of I-2Dm:PP1c complexes suggests that inhibition of PP1 activity in vivo does not result in a compensating increase in synthesis of active PP1. PP1 mutants enhance the wing overgrowth phenotype caused by ectopic expression of the type II TGF beta superfamily signaling receptor Punt. Using I-2Dm, which has a less severe effect than NIPP1Dm, we show that lowering the level of PP1 activity specifically in cells overexpressing Punt is sufficient for wing overgrowth and that the interaction between PP1 and Punt requires the type I receptor Thick-veins (Tkv) but is not strongly sensitive to the level of the ligand, Decapentaplegic (Dpp), nor to that of the other type I receptors. This is consistent with a role for PP1 in antagonizing Punt by preventing phosphorylation of Tkv. These studies demonstrate that inhibitors of PP1 can be used in a tissue- and developmental-specific manner to examine the developmental roles of PP1. PMID- 12750337 TI - Is the rate of insertion and deletion mutation male biased?: Molecular evolutionary analysis of avian and primate sex chromosome sequences. AB - The rate of mutation for nucleotide substitution is generally higher among males than among females, likely owing to the larger number of DNA replications in spermatogenesis than in oogenesis. For insertion and deletion (indel) mutations, data from a few human genetic disease loci indicate that the two sexes may mutate at similar rates, possibly because such mutations arise in connection with meiotic crossing over. To address origin- and sex-specific rates of indel mutation we have conducted the first large-scale molecular evolutionary analysis of indels in noncoding DNA sequences from sex chromosomes. The rates are similar on the X and Y chromosomes of primates but about twice as high on the avian Z chromosome as on the W chromosome. The fact that indels are not uncommon on the nonrecombining Y and W chromosomes excludes meiotic crossing over as the main cause of indel mutation. On the other hand, the similar rates on X and Y indicate that the number of DNA replications (higher for Y than for X) is also not the main factor. Our observations are therefore consistent with a role of both DNA replication and recombination in the generation of short insertion and deletion mutations. A significant excess of deletion compared to insertion events is observed on the avian W chromosome, consistent with gradual DNA loss on a nonrecombining chromosome. PMID- 12750338 TI - Genetic diversity and population history of golden monkeys (Rhinopithecus roxellana). AB - Golden monkey (Rhinopithecus roxellana), namely the snub-nosed monkey, is a well known endangered primate, which distributes only in the central part of mainland China. As an effort to understand the current genetic status as well as population history of this species, we collected a sample of 32 individuals from four different regions, which cover the major habitat of this species. Forty-four allozyme loci were surveyed in our study by allozyme electrophoresis, none of which was found to be polymorphic. The void of polymorphism compared with that of other nonhuman primates is surprising particularly considering that the current population size is many times larger than that of some other endangered species. Since many independent loci are surveyed in this study, the most plausible explanation for our observation is that the population has experienced a recent bottleneck. We used a coalescent approach to explore various scenarios of population bottleneck and concluded that the most recent bottleneck could have happened within the last 15,000 years. Moreover, the proposed simulation approach could be useful to researchers who need to analyze the non- or low-polymorphism data. PMID- 12750339 TI - Sry expression level and protein isoform differences play a role in abnormal testis development in C57BL/6J mice carrying certain Sry alleles. AB - Transfer of certain Mus domesticus-derived Y chromosomes (Sry(DOM) alleles, e.g., Sry(POS) and Sry(AKR)) onto the C57BL/6J (B6) mouse strain causes abnormal gonad development due to an aberrant interaction between the Sry(DOM) allele and the B6 derived autosomal (tda) genes. For example, B6 XY(POS) fetuses develop ovaries and ovotestes and B6 XY(AKR) fetuses have delayed testis cord development. To test whether abnormal testis development is caused by insufficient Sry(DOM) expression, two approaches were used. First, gonad development and relative Sry expression levels were examined in fetal gonads from two strains of B6 mice that contained a single M. domesticus-derived and a single M. musculus-derived Sry allele (B6-Y(POS,RIII) and B6-Y(AKR,RIII)). In both cases, presence of the M. musculus Sry(RIII) allele corrected abnormal testis development. On the B6 background, Sry(POS) was expressed at about half the level of Sry(RIII) whereas Sry(AKR) and Sry(RIII) were equally expressed. On an F(1) hybrid background, both Sry(POS) and Sry(RIII) expression increased, but Sry(POS) expression increased to a greater extent. Second, sexual development and Sry expression levels were determined in XX mice carrying a transgene expressing Sry(POS) controlled by POS derived or MUS-derived regulatory regions. In both cases one B6 transgenic line was recovered in which XX transgenic mice developed only testicular tissue but cord development was delayed despite normal Sry transcriptional initiation and overexpression. For three transgenes where B6 XX transgenic mice developed as females, hermaphrodites, or males, the percentage of XX transgenic males increased on an F(1) background. For the one transgene examined, Sry expression increased on an F(1) background. These results support a model in which delayed testis development is caused by the presence of particular DOM SRY protein isoforms and this, combined with insufficient Sry expression, causes sex reversal. These results also indicate that at least one tda gene regulates Sry expression, possibly by directly binding to Sry regulatory regions. PMID- 12750340 TI - Evolution of MHC class II E beta diversity within the genus Peromyscus. AB - Progress in understanding the evolution of variation at the MHC has been slowed by an inability to assess the relative roles of mutation vs. intragenic recombination in contributing to observed polymorphism. Recent theoretical advances now permit a quantitative treatment of the problem, with the result that the amount of recombination is at least an order of magnitude greater than that of mutation in the history of class II genes. We suggest that this insight allows progress in evaluating the importance of other factors affecting the evolution of the MHC. We investigated the evolution of MHC class II E beta sequence diversity in the genus Peromyscus. We find evidence for extensive recombination in the history of these sequences. Nevertheless, it appears that intragenic recombination alone is insufficient to account for evolution of MHC diversity in Peromyscus. Significant differences in silent variation among subgenera arose over a relatively short period of time, with little subsequent change. We argue that these observations are consistent with the effects of historical population bottleneck(s). Population restrictions may explain general features of MHC evolution, including the large amount of recombination in the history of MHC genes, because intragenic recombination may efficiently regenerate allelic polymorphism following a population constriction. PMID- 12750341 TI - Cloning and characterization of maize B chromosome sequences derived from microdissection. AB - Isolation of sequences from the maize B chromosome is always hampered by its high homology with the normal complements. In this study, this handicap was overcome by cloning the sequences from the pachytene B chromosomes dissected out of a slide by a micromanipulator followed by degenerate oligonucleotide-primed PCR. The isolated sequences were found to hybridize with genomic DNA in a B-dosage dependent manner and with the pachytene B chromosome by fluorescence in situ hybridization (FISH), corroborating their B origin. A total of 19 B sequences were isolated, all of which are repetitive and, with one exception, are homologous to the A chromosome(s). Three sequences have strong homology to maize sequences that include two knob repeats and one zein gene (noncoding region), and 10 others are homologous to the noncoding region of Adh1, Bz1, Gag, Zein, and B centromere to a lesser degree. Six sequences have no homology to any gene. In addition to FISH, the B-specific sequence and a partially B-specific one were also mapped, by seven newly characterized TB-10L translocations, to a similar location on the central portion of the distal heterochromatic region, spreading over a region of about one-third of the B chromosome. PMID- 12750342 TI - A bacterial artificial chromosome contig spanning the major domestication locus Q in wheat and identification of a candidate gene. AB - The Q locus played a major role in the domestication of wheat because it confers the free-threshing character and influences many other agronomically important traits. We constructed a physical contig spanning the Q locus using a Triticum monococcum BAC library. Three chromosome walking steps were performed by complete sequencing of BACs and identification of low-copy markers through similarity searches of database sequences. The BAC contig spans a physical distance of approximately 300 kb corresponding to a genetic distance of 0.9 cM. The physical map of T. monococcum had perfect colinearity with the genetic map of wheat chromosome arm 5AL. Recombination data in conjunction with analysis of fast neutron deletions confirmed that the contig spanned the Q locus. The Q gene was narrowed to a 100-kb segment, which contains an APETALA2 (AP2)-like gene that cosegregates with Q. AP2 is known to play a major role in controlling floral homeotic gene expression and thus is an excellent candidate for Q. PMID- 12750343 TI - A DNA-damage-induced cell cycle checkpoint in Arabidopsis. AB - Although it is well established that plant seeds treated with high doses of gamma radiation arrest development as seedlings, the cause of this arrest is unknown. The uvh1 mutant of Arabidopsis is defective in a homolog of the human repair endonuclease XPF, and uvh1 mutants are sensitive to both the toxic effects of UV and the cytostatic effects of gamma radiation. Here we find that gamma irradiation of uvh1 plants specifically triggers a G(2)-phase cell cycle arrest. Mutants, termed suppressor of gamma (sog), that suppress this radiation-induced arrest and proceed through the cell cycle unimpeded were recovered in the uvh1 background; the resulting irradiated plants are genetically unstable. The sog mutations fall into two complementation groups. They are second-site suppressors of the uvh1 mutant's sensitivity to gamma radiation but do not affect the susceptibility of the plant to UV radiation. In addition to rendering the plants resistant to the growth inhibitory effects of gamma radiation, the sog1 mutation affects the proper development of the pollen tetrad, suggesting that SOG1 might also play a role in the regulation of cell cycle progression during meiosis. PMID- 12750345 TI - Genetic analysis of early flowering mutants in Arabidopsis defines a class of pleiotropic developmental regulator required for expression of the flowering-time switch flowering locus C. AB - The Arabidopsis flowering-repressor gene FLOWERING LOCUS C (FLC) is a developmental switch used to trigger floral induction after extended growth in the cold, a process termed vernalization. In vernalized plants, FLC becomes transcriptionally silenced through a process that involves an epigenetic mechanism. We identified recessive mutations designated vernalization independence (vip) that confer cold-independent flowering and suppression of FLC. These mutations also lead to developmental pleiotropy, including specific defects in floral morphology, indicating that the associated genes also have functions unrelated to flowering time. We identified the VIP3 gene by positional cloning and found that it encodes a protein consisting almost exclusively of repeated Trp Asp (WD) motifs, suggesting that VIP3 could act as a platform to assemble a protein complex. Constitutive transgenic expression of VIP3 in vernalized plants is insufficient to activate FLC, and thus VIP3 probably participates in the regulation of FLC as one component of a more extensive mechanism. Consistent with this, genetic analyses revealed that the VIP loci define a functional gene class including at least six additional members. We suggest that VIP3 and other members of this gene class could represent a previously unrecognized flowering mechanism. PMID- 12750344 TI - The SHOOTLESS2 and SHOOTLESS1 genes are involved in both initiation and maintenance of the shoot apical meristem through regulating the number of indeterminate cells. AB - To characterize the SHL2 and SHL1 genes in detail, we analyzed three strains carrying weak alleles of SHL2, shl2-6, shl2-7, and shl2-8, and one weak allele of SHL1, shl1-3. In contrast to strong alleles, which result in lack of shoot meristem, strains bearing these weak alleles formed shoot meristem frequently during embryogenesis. In shl2-6 and shl2-7 mutants, the meristem was lost during seed development. Only the shl2-8 mutant could survive after germination, but it showed abnormal initiation pattern and morphology of leaves. In strains bearing the weak alleles, the shoot meristem was composed of a small number of indeterminate cells and ultimately converted into leaf primordium. The shl1-3 mutant showed phenotypes similar to those of shl2-8. Thus SHL2 and SHL1 are required for both initiation and maintenance of shoot meristem. In shl2 mutants, there was a positive correlation between the size of the expression domain of OSH1 representing the number of indeterminate cells, the frequency of shoot meristem initiation, and the duration of meristem survival. Thus the shoot meristem will not initiate in an "all-or-nothing" fashion, but is formed in various degrees depending on the strength of the alleles. Double-mutant analyses indicate that SHL2 functions upstream of SHO to establish proper organization of the shoot meristem. PMID- 12750346 TI - Comparison of a Brassica oleracea genetic map with the genome of Arabidopsis thaliana. AB - Brassica oleracea is closely related to the model plant, Arabidopsis thaliana. Despite this relationship, it has been difficult to both identify the most closely related segments between the genomes and determine the degree of genome replication within B. oleracea relative to A. thaliana. These difficulties have arisen in part because both species have replicated genomes, and the criteria used to identify orthologous regions between the genomes are often ambiguous. In this report, we compare the positions of sequenced Brassica loci with a known position on a B. oleracea genetic map to the positions of their putative orthologs within the A. thaliana genome. We use explicit criteria to distinguish orthologous from paralogous loci. In addition, we develop a conservative algorithm to identify collinear loci between the genomes and a permutation test to evaluate the significance of these regions. The algorithm identified 34 significant A. thaliana regions that are collinear with >28% of the B. oleracea genetic map. These regions have a mean of 3.3 markers spanning 2.1 Mbp of the A. thaliana genome and 2.5 cM of the B. oleracea genetic map. Our findings are consistent with the hypothesis that the B. oleracea genome has been highly rearranged since divergence from A. thaliana, likely as a result of polyploidization. PMID- 12750347 TI - Mathematical issues arising from the directed mutation controversy. AB - During the past 14 years or so a large body of new evidence that supposedly supports the directed mutation hypothesis has accumulated. Interpretation of some of the evidence depends on mathematical reasoning, which can be subtler than it appears at first sight. This article attempts to clarify some of the mathematical issues arising from the directed mutation controversy, thereby offering alternative interpretations of some of the evidence. PMID- 12750348 TI - Effects of population structure and admixture on exact tests for association between Loci. AB - The probability of multilocus genotype counts conditional on allelic counts and on allelic independence provides a test statistic for independence within and between loci. As the number of loci increases and each sampled genotype becomes unique, the conditional probability becomes a function of total heterozygosity. In that case, it does not address between-locus dependence directly but only indirectly through detection of the Wahlund effect. Moreover, the test will reject the hypothesis of allelic independence only for small values of heterozygosity. Low heterozygosity is expected for population subdivision but not for population admixture. The test may therefore be inappropriate for admixed populations. If individuals with parents in two different populations are always considered to belong to one of the populations, then heterozygosity is increased in that population and the exact test should not be used for sparse data sets from that population. If such a case is suspected, then alternative testing strategies are suggested. PMID- 12750349 TI - Selective sweeps in the presence of interference among partially linked loci. AB - Recurrent directional selection on a partially recombining chromosome may cause a substantial reduction of standing genetic variation in natural populations. Previous studies of this effect, commonly called selective sweeps, assumed that at most one beneficial allele is on the way to fixation at a given time. However, for a high rate of selected substitutions and a low recombination rate, this assumption can easily be violated. We investigated this problem using full forward simulations and analytical approximations. We found that interference between linked beneficial alleles causes a reduction of their fixation probabilities. The hitchhiking effect on linked neutral variation for a given substitution also slightly decreases due to interference. As a result, the strength of recurrent selective sweeps is weakened. However, this effect is significant only in chromosomal regions of relatively low recombination rates where the level of variation is greatly reduced. Therefore, previous results on recurrent selective sweeps although derived for a restricted parameter range are still valid. Analytical approximations are obtained for the case of complete linkage for which interference between competing beneficial alleles is maximal. PMID- 12750351 TI - A coalescent model of recombination hotspots. AB - Recent experimental findings suggest that the assumption of a homogeneous recombination rate along the human genome is too naive. These findings point to block-structured recombination rates; certain regions (called hotspots) are more prone than other regions to recombination. In this report a coalescent model incorporating hotspot or block-structured recombination is developed and investigated analytically as well as by simulation. Our main results can be summarized as follows: (1) The expected number of recombination events is much lower in a model with pure hotspot recombination than in a model with pure homogeneous recombination, (2) hotspots give rise to large variation in recombination rates along the genome as well as in the number of historical recombination events, and (3) the size of a (nonrecombining) block in the hotspot model is likely to be overestimated grossly when estimated from SNP data. The results are discussed with reference to the current debate about block-structured recombination and, in addition, the results are compared to genome-wide variation in recombination rates. A number of new analytical results about the model are derived. PMID- 12750350 TI - Power evaluations for family-based tests of association with incomplete parental genotypes. AB - While a variety of methods have been developed to deal with incomplete parental genotype information in family-based association tests, sampling design issues with incomplete parental genotype data still have not received much attention. In this article, we present simulation studies with four genetic models and various sampling designs and evaluate power in family-based association studies. Efficiency depends heavily on disease prevalence. With rare diseases, sampling affecteds and their parents is preferred, and three sibs will be required to have close power if parents are unavailable. With more common diseases, sampling affecteds and two sibs will generally be more efficient than trios. When parents are unavailable, siblings need not be phenotyped if the disease is rare, but a loss of power will result with common diseases. Finally, for a class of complex traits where other genetic and environmental factors also cause phenotypic correlation among siblings, little loss of efficiency occurs to rare disease, but substantial loss of efficiency occurs to common disease. PMID- 12750352 TI - Autoimmune kidney disease and lymphadenopathy in NODlpr mice are not modified by deficiency in tumor necrosis factor receptor 1 or beta 2-microglobulin. AB - Fas and TNFRI, two members of the tumor necrosis factor receptor family with an intracellular death domain, each play critical roles in apoptotic death of lymphocytes and certain other cell types. We determined the overlapping functions of Fas and TNFRI by breeding non-obese diabetic (NOD) mutant mice that lacked both receptors. NODlpr mice developed extensive lymphadenopathy, splenomegaly, CD4(-)CD8(-) B220(+) alpha beta TCR(+) T cells and autoimmune kidney disease. This pathology was not modified by concomitant deficiency in TNFRI as was reported for lpr mice on a B6 background. NODlpr mice lacking CD8(+) T cells, because of a null mutation in beta(2)-microglobulin (beta(2)m), also developed a similar disease profile to NODlpr animals, but the CD4(-)CD8(-) B220(+) alpha beta TCR(+) T cells now derived from a CD4(+) T cell lineage. These results demonstrate that, as in the autoimmune-prone MRL stain, the NOD genetic background promotes lupus nephritis-like pathology and extensive lymphadenopathy when lpr is present. Loss of TNFRI does not exacerbate the pathology caused by deficiency in Fas and loss of beta(2)m does not reduce it. PMID- 12750353 TI - Engineering Th determinants for efficient priming of humoral and cytotoxic T cell responses. AB - To engineer T(h) determinants (THd) to prime help for humoral or cytotoxic T cell responses, we modified ovalbumin [OVA(323-337)] and myoglobin [MYO(106-118)] eliciting T(h)1 and T(h)0 cytokine profiles respectively. Residues along the sequence of both THd were replaced with amino acids representative of different families. Replacements at positions P-1 and P5 pointing to the TCR in both THd afforded higher levels of IFN-gamma and IL-4 production. Peptides eliciting different proportions of IFN-gamma and IL-4 were co-immunized with a peptide hapten or a T cytotoxic determinant (TCd) respectively. OVA(323-337)- and MYO(106 118)-derived peptides afforded the best THd for the induction of cytotoxic T lymphocyte (CTL) and anti-hapten antibodies respectively. IFN-gamma and IL-4, primed by MYO(106-118)-derived peptides, correlated significantly with antibody production against the hapten (P < 0.05 for IFN-gamma and P < 0.05 for IL-4). Interestingly, two peptides derived from OVA(323-337), 323G and 327G, which induced the clearest T(h)2 cytokine profiles, were not the most efficient to prime cell help for the induction of anti-hapten antibodies. For CTL induction, OVA(323-337)-derived peptides, inducing a T(h)1-like profile, required a lower dose (5 nmol) than T(h)0 peptides (50 nmol). The dose of 50 nmol was detrimental for T(h)1-like peptides. Interestingly, IFN-gamma primed by the THd correlated significantly with that induced by the TCd (P < 0.01). PMID- 12750354 TI - Differential regulation of VLA-2 expression on Th1 and Th2 cells: a novel marker for the classification of Th subsets. AB - We found that T(h)1 cells derived from ovalbumin (OVA)-specific TCR transgenic (DO11.10) mice showed significantly higher levels of VLA-2 (CD49b/CD29) expression than T(h)2 cells. In the early days (until 6 days) during induction of T(h)1 or T(h)2 cells, the expression of VLA-2 was gradually increased on both T(h) subsets. Thereafter, VLA-2 expression was further up-regulated on T(h)1 cells until 13 days, while a significant decrease of VLA-2 was observed in T(h)2 cells, resulting in a marked difference of expression at day 13. Up-regulation of VLA-2 on T(h)1 cells was not impaired in IFN-gamma(-/-) T(h) cells nor blocked by anti-IL-12 mAb treatment on wild-type T(h) cells, suggesting that up-regulation of VLA-2 on T(h)1 cells occurs in an IFN-gamma- and IL-12-independent manner. In contrast, T(h) cells cultured under IL-4-depleted T(h)2 conditions abrogated the down-regulation of VLA-2 expression, suggesting that down-regulation of VLA-2 expression on T(h)2 cells was dependent on IL-4. The finding that STAT6(-/-) T(h)2 cells did not show any down-regulation of VLA-2 expression and expressed the same levels of VLA-2 as T(h)1 cells indicated a critical role for the IL-4 receptor/STAT6 signaling pathway in IL-4-dependent down-regulation of VLA-2 on T(h)2 cells. Stimulation of T(h)1 cells by VLA-2 ligands such as collagen type I or agonistic mAb provided co-stimulation for anti-CD3 mAb-induced IFN-gamma production. However, these ligations had little effect on the IL-4 production of T(h)2 cells. Together, these results indicate that VLA-2 is a novel functional marker that dissociates T(h)1 from T(h)2 cells, and thus might be useful for therapeutic monitoring of T(h)1-dependent immune diseases such as rheumatoid arthritis or Crohn's disease. PMID- 12750355 TI - A role for dendritic cells in the priming of antigen-specific CD4+ and CD8+ T lymphocytes by immune-stimulating complexes in vivo. AB - Immune-stimulating complexes (ISCOMS) are adjuvant vectors which are unusual in being able to prime both CD4(+) and CD8(+) T cells by parenteral and mucosal routes. However, their mode of action is unclear and to define better the cellular interactions involved we have studied the ability of ISCOMS containing ovalbumin (OVA) to prime TCR transgenic CD4(+) or CD8(+) T cells in vivo. Immunization with OVA ISCOMS caused activation and clonal expansion of CD4(+) and CD8(+) T cells in the T cell areas of the draining lymph nodes, followed by the migration of both CD4(+) and CD8(+) T cells into the B cell follicle. The T cells were primed to proliferate and secrete IFN-gamma after re-stimulation in vitro with the appropriate OVA peptide and CD8(+) T cell priming occurred in the absence of CD4(+) T cells. Increasing the number of dendritic cells (DC) in vivo with flt3 ligand augmented the expansion and activation of the OVA-specific T cells, particularly CD8(+) T cells. These studies indicate DC play a central role in the priming of both CD4(+) and CD8(+) T cells in vivo, and suggest that an ability to target DC may allow ISCOMS to be powerful vaccine vectors for stimulating protective immunity. PMID- 12750356 TI - Expression and function of Toll-like receptors 2 and 4 in human keratinocytes. AB - Keratinocytes have the ability to kill pathogenic fungi and bacteria by producing antimicrobial substances. Recent studies suggest that microbial components use signaling molecules of the human Toll-like receptor (TLR) family to transduce signals in various cells. Here we provide evidence that keratinocytes express both TLR2 and TLR4 at the mRNA and protein levels, and show that TLR2 and TLR4 are present in the normal human epidermis in vivo and that their expression is regulated by microbial components. The expression of myeloid differentiation protein gene (MyD88), which is involved in the signaling pathway of many TLR, was also demonstrated in keratinocytes. LPS + IFN-gamma increased the expression of TLR2 and TLR4 50- and 5-fold respectively. Treatment of keratinocytes with Candida albicans, mannan, Mycobacterium tuberculosis or LPS with IFN-gamma resulted in the activation and nuclear translocation of NF-kappaB. Inhibition of NF-kappaB blocked the Candida-killing activity of keratinocytes, suggesting that the antimicrobial effect of keratinocytes requires NF-kappaB activation. LPS + IFN-gamma, C. albicans (4 Candida/KC), peptidoglycan (1 micro g/ml) or M. tuberculosis extract significantly increased IL-8 gene expression after 3 h of treatment (P < 0.05). The increases over the 0-h level were 15-, 8-, 10.8- and 7 fold, respectively. The microbial compound-induced increase in IL-8 gene expression could be inhibited by anti-TLR2 and anti-TLR4 neutralizing antibodies, suggesting that TLRs are involved in the pathogen-induced expression of this pro inflammatory cytokine. Our findings stress the importance of the role of keratinocytes as a component of innate immunity. PMID- 12750357 TI - Nasal tolerance induces antigen-specific CD4+CD25- regulatory T cells that can transfer their regulatory capacity to naive CD4+ T cells. AB - The mucosal immune system is uniquely adapted to elicit immune responses against pathogens but also to induce tolerogenic responses to harmless antigens. In mice, nasal application of ovalbumin (OVA) leads to suppression of both T(h)1 and T(h)2 responses. This tolerance can be transferred to naive mice by CD4(+) T(r) cells from the spleen. Using the allotypic Ly5 system, we were able to demonstrate in vivo that T(r) cells not only suppress naive CD4(+) T cells, but also induce them to differentiate into T(r) cells. The effector function of these mucosal T(r) cells is not restricted by cytokine polarization, since T(r) cells from T(h)1 tolerant mice can suppress a T(h)2 response and vice versa. Transfer of splenic CD4(+)CD25(+) and CD4(+)CD25(-) T cell subsets from OVA-tolerized mice revealed that both subsets were equally able to suppress a delayed-type hypersensitivity response in acceptor mice. In contrast to the CD25(-) T cell subset, the CD25(+) cells were not specific for the antigen used for tolerization. Together, these findings demonstrate a role for CD4(+)CD25(-) T(r) cells in mucosal tolerance, which suppresses CD4(+) T cells in an antigen-specific fashion, irrespective of initial T(h)1/T(h)2 skewing of the immune response. This offers a major advantage in the manipulation of mucosal tolerance for the treatment of highly cytokine polarized disorders such as asthma and autoimmune diseases. PMID- 12750358 TI - Critical function of T cell death-associated gene 8 in glucocorticoid-induced thymocyte apoptosis. AB - Transcriptional expression of a gene or genes is absolutely required for induction of glucocorticoid-induced thymocyte apoptosis. We have previously shown that expression of T cell death-associated gene 8 (TDAG8) is quickly induced exclusively in the thymus after dexamethasone (DEX) treatment. Here, we present data that TDAG8 expression is induced prior to induction of DEX-mediated apoptosis. In contrast, TDAG8 expression in thymocytes was not induced in the process of gamma-irradiation-mediated apoptosis. TDAG8 expression accelerated only DEX-induced, but not TCR-mediated or gamma-irradiation-induced, thymocyte apoptosis in transgenic mice overexpressing TDAG8. Interestingly, these effects were specifically detected in CD4(+)CD8(+) double-positive thymocytes. Moreover, activation of caspase-3, -8 and -9 was enhanced in thymocytes of TDAG8 transgenic mice after DEX stimulation. In conclusion, TDAG8 expression is involved in glucocorticoid-induced signals to activate caspase-9, -8 and -3 for subsequent apoptosis induction in CD4(+)CD8(+) double-positive thymocytes. PMID- 12750359 TI - Naturally occurring peptides associated with HLA-A2 in ovarian cancer cell lines identified by mass spectrometry are targets of HLA-A2-restricted cytotoxic T cells. AB - Identifying naturally occurring peptides bound to HLA class I molecules recognized by HLA-restricted cytotoxic T lymphocytes (CTL) is both relevant and central to the development of effective immunotherapeutic strategies against cancer. Several cancer-related genes have been reported for ovarian cancer, but very few are known to be naturally processed T cell epitopes. In the present study we used mass spectrometry to identify 16 novel HLA-A2-bound peptides from HLA-A2(+) ovarian cancer cell lines. All 16 peptides are derived from source proteins with diverse functions and marked homology to known proteins found in public databases. Synthetic peptide analogues of identified sequences were found to stabilize HLA-A2.1, albeit with varying affinities. The peptides were found to be antigenic in that a primary CD8(+) CTL response could be elicited from normal donor blood. The CTL generated were not only peptide specific, but failed to recognize targets pulsed with control peptides. In addition, recognition of shared HLA-A2-restricted epitopes by these CTL is suggested by their reactivity with a subset of HLA-A2(+) tumor lines and freshly isolated cancer cells or cell lines established from peritoneal ascites. These results were further corroborated by competitive inhibition of lysis of an otherwise susceptible cell line in the presence of cold peptide-pulsed targets. Furthermore, lack of recognition of several HLA-A2(+) control cell lines or cells isolated from normal ovaries suggests that these peptides are cancer related. These findings broaden the list of CTL-defined antigens that could lead to the development of multi epitope vaccines for the treatment of ovarian cancer. PMID- 12750360 TI - HLA-B*0702 transgenic, H-2KbDb double-knockout mice: phenotypical and functional characterization in response to influenza virus. AB - HLA-B*0702 transgenic mice (expressing a chimeric heavy chain with a murine alpha 3 domain: HLA-B7(m alpha 3)) in which the H-2K(b) and H-2D(b) class I-a (Cl I-a( /-)) genes have been inactivated were compared with H-2K(b)D(b) Cl I-a(+/+) positive controls. Expression of the HLA-B7(m alpha 3) molecules resulted in a 3- to 4-fold increase in peripheral CD8(+) T lymphocyte numbers compared to H-2 Cl I a(-/-) knockout mice. These cells show a diversified TCR repertoire. Following influenza infection, a significant improvement in HLA-B0702-restricted cytotoxic T lymphocyte (CTL) responses was observed in HLA-B7(m alpha 3), H-2 Cl I-a(-/-) compared to HLA-B7(m alpha 3), H-2 Cl I-a(+/+) mice. The CTL response of infected HLA-B7(m alpha 3), H-2 Cl I-a(-/-) mice was directed against the nucleoprotein (NP) 418-426 epitope in which mutations have accumulated. Whereas all NP 418-426 variant peptides induced a CTL response, cross-reactivity to the variants was affected. These NP mutations could have been selected over time in humans for the virus to escape HLA-B0702-restricted CTL responses since a similar response was seen in humans with, as in mice, altered cross-recognition of the NP 418-426 variants. These animals may prove a suitable model to study HLA-B0702-restricted CTL responses. PMID- 12750361 TI - Distinct antigen trafficking from skin in the steady and active states. AB - In antigen trafficking from the skin, it has been postulated that Langerhans cells/dendritic cells are activated after capturing exogenous antigens, up regulate the expression of the chemokine receptor, CCR7, and migrate into lymphoid organs in response to the signaling of a chemokine, CCL21, which is expressed in lymphatic vessels and T cell zone stromal cells. Here we demonstrate that there is a distinct pathway of antigen trafficking from skin in the steady state that is independent of CCL21-CCR7 signaling. Employing melanin granules as an endogenous traceable antigen, we developed a system for visualizing antigen trafficking using mice with melanocytosis in the skin. We found the abrogation of antigen trafficking into regional lymph nodes (LN) in CCL21-Ser-deficient paucity of lymph node T cells (plt) mice in the active state induced by lipopolysaccharide injection, corresponding with previous reports, but normal accumulation of antigen in regional LN under steady-state conditions. These findings suggest that self-antigen is trafficking constitutively using pathway(s) other than that of the active state and the constitutive trafficking might regulate self-reactivity of the immune system. PMID- 12750362 TI - Evidence for a role of MCM (mini-chromosome maintenance)5 in transcriptional repression of sub-telomeric and Ty-proximal genes in Saccharomyces cerevisiae. AB - The MCM (mini-chromosome maintenance) genes have a well established role in the initiation of DNA replication and in the elongation of replication forks in Saccharomyces cerevisiae. In this study we demonstrate elevated expression of sub telomeric and Ty retrotransposon-proximal genes in two mcm5 strains. This pattern of up-regulated genes resembles the genome-wide association of MCM proteins to chromatin that was reported earlier. We link the altered gene expression in mcm5 strains to a reversal of telomere position effect (TPE) and to remodeling of sub telomeric and Ty chromatin. We also show a suppression of the Ts phenotype of a mcm5 strain by the high copy expression of the TRA1 component of the chromatin remodeling SAGA/ADA (SPT-ADA-GCN5 acetylase/ADAptor). We propose that MCM proteins mediate the establishment of silent chromatin domains around telomeres and Ty retrotransposons. PMID- 12750363 TI - Redox-Bohr and other cooperativity effects in the nine-heme cytochrome C from Desulfovibrio desulfuricans ATCC 27774: crystallographic and modeling studies. AB - The nine-heme cytochrome c is a monomeric multiheme cytochrome found in Desulfovibrio desulfuricans ATCC 27774. The polypeptide chain comprises 296 residues and wraps around nine hemes of type c. It is believed to take part in the periplasmic assembly of proteins involved in the mechanism of hydrogen cycling, receiving electrons from the tetraheme cytochrome c3. With the purpose of understanding the molecular basis of electron transfer processes in this cytochrome, we have determined the crystal structures of its oxidized and reduced forms at pH 7.5 and performed theoretical calculations of the binding equilibrium of protons and electrons in these structures. This integrated study allowed us to observe that the reduction process induced relevant conformational changes in several residues, as well as protonation changes in some protonatable residues. In particular, the surroundings of hemes I and IV constitute two areas of special interest. In addition, we were able to ascertain the groups involved in the redox Bohr effect present in this cytochrome and the conformational changes that may underlie the redox-cooperativity effects on different hemes. Furthermore, the thermodynamic simulations provide evidence that the N- and C-terminal domains function in an independent manner, with the hemes belonging to the N-terminal domain showing, in general, a lower redox potential than those found in the C terminal domain. In this way, electrons captured by the N-terminal domain could easily flow to the C-terminal domain, allowing the former to capture more electrons. A notable exception is heme IX, which has low redox potential and could serve as the exit path for electrons toward other proteins in the electron transfer pathway. PMID- 12750365 TI - Agonist-induced formation of opioid receptor-G protein-coupled receptor kinase (GRK)-G beta gamma complex on membrane is required for GRK2 function in vivo. AB - G protein-coupled receptor kinases (GRKs) catalyze agonist-induced receptor phosphorylation on the membrane and initiate receptor desensitization. Previous in vitro studies have shown that the binding of GRK to membrane-associated G beta gamma subunits plays an important role in translocation of GRK2 from the cytoplasm to the plasma membrane. The current study investigated the role of the interaction of GRK2 with the activated delta-opioid receptor (DOR) and G beta gamma subunits in the membrane translocation and function of GRK2 using intact human embryonic kidney 293 cells. Our results showed that agonist treatment induced GRK2 binding to DOR, GRK2 translocation to the plasma membrane, and DOR phosphorylation in cells expressing the wild-type DOR but not the mutant DOR lacking the carboxyl terminus, which contains all three GRK2 phosphorylation sites. DORs with the GRK2 phosphorylation sites modified (M3) or with the acidic residues flanking phosphorylation sites mutated (E355Q/D364N) failed to be phosphorylated in response to agonist stimulation. Agonist-induced GRK2 membrane translocation and GRK-receptor association were observed in cells expressing M3 but not E355Q/D364N. Moreover, over-expression of G beta gamma subunits promoted GRK2 binding to DOR, whereas over-expression of transducin alpha or the carboxyl terminus of GRK2 blocked binding. Further study demonstrated that agonist stimulation induced the formation of a complex containing DOR, GRK2, and G beta gamma subunits in the cell and that agonist-stimulated formation of this complex is essential for the stable localization of GRK2 on the membrane and for its catalytic activity in vivo. PMID- 12750364 TI - Entrapment of Rho ADP-ribosylated by Clostridium botulinum C3 exoenzyme in the Rho-guanine nucleotide dissociation inhibitor-1 complex. AB - RhoA, -B, and -C are ADP-ribosylated by Clostridium botulinum exoenzyme C3 to induce redistribution of the actin filaments in intact cells, a finding that has led to the notion that the ADP-ribosylation blocks coupling of Rho to the downstream effectors. ADP-ribosylation, however, does not alter nucleotide binding, intrinsic, and GTPase-activating protein-stimulated GTPase activity. ADP ribosylated Rho is even capable of activating the effector protein ROK in a recombinant system. Treatment of cells with a cell-permeable chimeric C3 toxin led to complete localization of modified Rho to the cytosolic fraction based on the complexation of ADP-ribosylated Rho with the guanine-nucleotide dissociation inhibitor-1 (GDI-1). The modified complex turned out to be resistant to phosphatidylinositol 4,5-bisphosphate- and GTPgammaS-induced release of Rho from GDI-1. Thus, ADP-ribosylation leads to entrapment of Rho in the GDI-1 complex. The increased stability of the GDI complex prevented binding of Rho to membrane associated players of the GTPase cycle such as the activating guanine nucleotide exchange factors and effector proteins. PMID- 12750366 TI - Identification of genes encoding arabinosyltransferases (SCA) mediating developmental modifications of lipophosphoglycan required for sand fly transmission of leishmania major. AB - At key steps in the infectious cycle pathogens must adhere to target cells, but at other times detachment is required for transmission. During sand fly infections by the protozoan parasite Leishmania major, binding of replicating promastigotes is mediated by galactosyl side chain (scGal) modifications of phosphoglycan repeats of the major surface adhesin, lipophosphoglycan (LPG). Release is mediated by arabinosyl (Ara) capping of LPG scbetaGal residues upon differentiation to the infective metacyclic stage. We used intraspecific polymorphisms of LPG structure to develop a genetic strategy leading to the identification of two genes (SCA1/2) mediating scAra capping. These LPG side chain beta1,2-arabinosyltransferases (scbetaAraTs) exhibit canonical glycosyltransferase motifs, and their overexpression leads to elevated microsomal scbetaAraT activity. Although the level of scAra caps is maximal in metacyclic parasites, scbetaAraT activity is maximal in log phase cells. Because quantitative immunolocalization studies suggest this is not mediated by sequestration of SCA scbetaAraTs away from the Golgi apparatus during log phase, regulation of activated Ara precursors may control LPG arabinosylation in vivo. The SCA genes define a new family of eukaryotic betaAraTs and represent novel developmentally regulated LPG-modifying activities identified in Leishmania. PMID- 12750367 TI - Bis(glutathionyl)spermine and other novel trypanothione analogues in Trypanosoma cruzi. AB - Trypanosomatids differ from other cells in their ability to conjugate glutathione with the polyamine spermidine to form the antioxidant metabolite trypanothione (N1,N8-bis(glutathionyl)spermidine). In Trypanosoma cruzi, trypanothione is synthesized by an unusual trypanothione synthetase/amidase (TcTryS) that forms both glutathionylspermidine and trypanothione. Because T. cruzi is unable to synthesize putrescine and is dependent on uptake of exogenous polyamines by high affinity transporters, synthesis of trypanothione may be circumstantially limited by lack of spermidine. Here, we show that the parasite is able to circumvent the potential shortage of spermidine by conjugating glutathione with other physiological polyamine substrates from exogenous sources (spermine, N8 acetylspermidine, and N-acetylspermine). Novel thiols were purified from epimastigotes, and structures were determined by matrix-assisted laser desorption ionization time-of-flight analysis to be N1,N12-bis(glutathionyl)spermine, N1 glutathionyl-N8-acetylspermidine, and N1-glutathionyl-N12-acetylspermine, respectively. Structures were confirmed by enzymatic synthesis with recombinant TcTryS, which catalyzes formation of these compounds with kinetic parameters equivalent to or better than those of spermidine. Despite containing similar amounts of spermine and spermidine, the epimastigotes, trypomastigotes, and amastigotes of T. cruzi preferentially synthesized trypanothione. Bis(glutathionyl)spermine disulfide is a physiological substrate of recombinant trypanothione reductase, comparable to trypanothione and homotrypanothione disulfides. The broad substrate specificity of TcTryS could be exploited in the design of polyamine-based inhibitors of trypanothione metabolism. PMID- 12750368 TI - The cell cycle-regulated protein human GTSE-1 controls DNA damage-induced apoptosis by affecting p53 function. AB - GTSE-1 (G2 and S phase-expressed-1) protein is specifically expressed during S and G2 phases of the cell cycle. It is mainly localized to the microtubules and when overexpressed delays the G2 to M transition. Here we report that human GTSE 1 (hGTSE-1) protein can negatively regulate p53 transactivation function, protein levels, and p53-dependent apoptosis. We identified a physical interaction between the C-terminal regulatory domain of p53 and the C-terminal region of hGTSE-1 that is necessary and sufficient to down-regulate p53 activity. Furthermore, we provide evidence that hGTSE-1 is able to control p53 function in a cell cycle dependent fashion. hGTSE-1 knock-down by small interfering RNA resulted in a S/G2 specific increase of p53 levels as well as cell sensitization to DNA damage induced apoptosis during these phases of the cell cycle. Altogether, this work suggests a physiological role of hGTSE-1 in apoptosis control after DNA damage during S and G2 phases through regulation of p53 function. PMID- 12750369 TI - The stable isotope-based dynamic metabolic profile of butyrate-induced HT29 cell differentiation. AB - Stable isotope-based dynamic metabolic profiling is applied in this paper to elucidate the mechanism by which butyrate induces cell differentiation in HT29 cells. We utilized butyrate-sensitive (HT29) cells incubated with [1,2 13C2]glucose or [1,2-13C2]butyrate as single tracers to observe the changes in metabolic fluxes in these cells. In HT29 cells, increasing concentrations of butyrate inhibited glucose uptake, glucose oxidation, and nucleic acid ribose synthesis in a dose-dependent fashion. Glucose carbon utilization for de novo fatty acid synthesis and tricarboxylic acid cycle flux was replaced by butyrate. We also demonstrated that these changes are not present in butyrate-resistant pancreatic adenocarcinoma MIA cells. The results suggest that the mechanism by which colon carcinoma cells acquire a differentiated phenotype is through a replacement of glucose for butyrate as the main carbon source for macromolecule biosynthesis and energy production. This provides a better understanding of cell differentiation through metabolic adaptive changes in response to butyrate in HT29 cells, demonstrating that variations in metabolic pathway substrate flow are powerful regulators of tumor cell proliferation and differentiation. PMID- 12750370 TI - Flexible DNA binding of the BTB/POZ-domain protein FBI-1. AB - POZ-domain transcription factors are characterized by the presence of a protein protein interaction domain called the POZ or BTB domain at their N terminus and zinc fingers at their C terminus. Despite the large number of POZ-domain transcription factors that have been identified to date and the significant insights that have been gained into their cellular functions, relatively little is known about their DNA binding properties. FBI-1 is a BTB/POZ-domain protein that has been shown to modulate HIV-1 Tat trans-activation and to repress transcription of some cellular genes. We have used various viral and cellular FBI 1 binding sites to characterize the interaction of a POZ-domain protein with DNA in detail. We find that FBI-1 binds to inverted sequence repeats downstream of the HIV-1 transcription start site. Remarkably, it binds efficiently to probes carrying these repeats in various orientations and spacings with no particular rotational alignment, indicating that its interaction with DNA is highly flexible. Indeed, FBI-1 binding sites in the adenovirus 2 major late promoter, the c-fos gene, and the c-myc P1 and P2 promoters reveal variously spaced direct, inverted, and everted sequence repeats with the consensus sequence G(A/G)GGG(T/C)(C/T)(T/C)(C/T) for each repeat. PMID- 12750371 TI - Cyclosporin A-insensitive permeability transition in brain mitochondria: inhibition by 2-aminoethoxydiphenyl borate. AB - The mitochondrial permeability transition pore (PTP) may operate as a physiological Ca2+ release mechanism and also contribute to mitochondrial deenergization and release of proapoptotic proteins after pathological stress, e.g. ischemia/reperfusion. Brain mitochondria exhibit unique PTP characteristics, including relative resistance to inhibition by cyclosporin A. In this study, we report that 2-aminoethoxydiphenyl borate blocks Ca2+-induced Ca2+ release in isolated, non-synaptosomal rat brain mitochondria in the presence of physiological concentrations of ATP and Mg2+. Ca2+ release was not mediated by the mitochondrial Na+/Ca2+ exchanger or by reversal of the uniporter responsible for energy-dependent Ca2+ uptake. Loss of mitochondrial Ca2+ was accompanied by release of cytochrome c and pyridine nucleotides, indicating an increase in permeability of both the inner and outer mitochondrial membranes. Under these conditions, Ca2+-induced opening of the PTP was not blocked by cyclosporin A, antioxidants, or inhibitors of phospholipase A2 or nitric-oxide synthase but was abolished by pretreatment with bongkrekic acid. These findings indicate that in the presence of adenine nucleotides and Mg2+,Ca2+-induced PTP in non-synaptosomal brain mitochondria exhibits a unique pattern of sensitivity to inhibitors and is particularly responsive to 2-aminoethoxydiphenyl borate. PMID- 12750372 TI - c-Src is activated by the epidermal growth factor receptor in a pathway that mediates JNK and ERK activation by gonadotropin-releasing hormone in COS7 cells. AB - Key participants in G protein-coupled receptor (GPCR) signaling are the mitogen activated protein kinase (MAPK) signaling cascades. The mechanisms involved in the activation of the above cascades by GPCRs are not fully elucidated. A prototypic GPCR that has been widely used to study these signaling mechanisms is the receptor for gonadotropin-releasing hormone (GnRHR), which serves as a key regulator of the reproductive system. Here we expressed GnRHR in COS7 cells and found that GnRHR transmits its signals to MAPKs mainly via G alpha i, EGF receptor without the involvement of Hb-EGF, and c-Src, but independently of PKCs. The main pathway that leads to JNK activation downstream of the EGF receptor involves a sequential activation of c-Src and phosphatidylinositol 3-kinase (PI3K). ERK activation by GnRHR is mediated by the EGF receptor, which activates Ras either directly or via c-Src. Besides the main pathway, the dissociated G beta gamma and beta-arrestin may initiate additional, albeit minor, pathways that lead to MAPK activation in the transfected COS7 cells. The pathways detected are significantly different from those in other cell lines bearing GnRHR, indicating that GnRH can utilize various signaling mechanisms for the activation of MAPK cascades. The unique pathway elucidated here in which c-Src and PI3K are sequentially activated downstream of the EGF receptor may serve as a prototype of signaling mechanisms by GnRHR and by additional GPCRs in various cell types. PMID- 12750373 TI - Substrate-induced conformational fit and headpiece closure in the Ca2+ATPase (SERCA). AB - Protection of the Ca2+ATPase (SERCA) from proteinase K digestion has been observed following the addition of Ca2+, Mg2+, and nucleotide and interpreted as a substrate-dependent conformational change (1). The protected digestion site is located on the loop connecting the A domain and the M3 transmembrane helix. We studied by mutational analysis the protective effect of AMP-PCP, an ATP analog that is not utilized for enzyme phosphorylation. We found that the nucleotide protective effect is interfered with by single mutations of Arg-560 and Glu-439 in the N domain and Lys-352, Lys-684, Thr-353, Asp-703, and Asp-707 in the P domain. This is consistent with a transition from the open to the compact configuration of the ATPase headpiece and approximation of the N and P domains by interactions with the nucleotide adenosine and phosphate moieties, respectively. The A domain-M3 loop is consequently involved. Protection by nucleotide substrate increased following the mutations of Asp-351 (the residue undergoing phosphorylation by ATP) and neighboring Asn-706 to Ala, underlying the importance of side chain specificity in positioning the nucleotide terminal phosphate and limiting the stability of the substrate-enzyme complex. Protection is not observed when AMP-PCP is added in the absence of Ca2+ or following mutations (E771Q or N796A) that interfere with Ca2+ binding. Therefore, nucleotide binds to the Ca2+-activated enzyme in the open headpiece conformation and the consequent approximation of the N and P domains occurs while the transmembrane domain is still in the Ca2+-bound conformation. Mg2+ is not required for the protective effect of nucleotide, even though it is specifically required for the subsequent catalytic reactions. PMID- 12750374 TI - Cell surface expression of 5-hydroxytryptamine type 3 receptors is controlled by an endoplasmic reticulum retention signal. AB - Two subunits of the 5-hydroxytryptamine type 3 (5-HT3) have been identified (5 HT3A and 5-HT3B) that assemble into homomeric (5-HT3A) and heteromeric (5-HT3A+5 HT3B) complexes. Unassembled 5-HT3B subunits are efficiently retained within the cell. In this study, we address the mechanism controlling the release of 5-HT3B from the endoplasmic reticulum (ER). An analysis of chimeric 5-HT3A receptor(R).5 HT3BR constructs suggests the presence of elements downstream of the first transmembrane domain of 5-HT3B subunits that inhibit cell surface expression. To investigate this possibility, truncated 5-HT3B subunits were constructed and assessed for their ability to access the cell surface in COS-7 and ts201 cells. Using this approach, we have identified the presence of an ER retention signal located within the first cytoplasmic loop between transmembrane domains I and II of 5-HT3B. Transplantation of this signal (CRAR) into the homologous region of 5 HT3A results in the ER retention of this subunit until rescued by co-assembly with wild-type 5-HT3A. The mutation of this ER retention signal in 5-HT3B (5 HT3BSGER) does not lead to cell surface expression, suggesting the presence of other signals or mechanisms to control the surface expression of 5-HT3BRs. The generation of truncated 5-HT3BSGER constructs confirmed that the CRAR signal does play an important role in the ER retention of 5-HT3B. PMID- 12750375 TI - Random mutagenesis of the M3 muscarinic acetylcholine receptor expressed in yeast. Identification of point mutations that "silence" a constitutively active mutant M3 receptor and greatly impair receptor/G protein coupling. AB - The M3 muscarinic receptor is a prototypical member of the class I family of G protein-coupled receptors (GPCRs). To facilitate studies on the structural mechanisms governing M3 receptor activation, we generated an M3 receptor expressing yeast strain (Saccharomyces cerevisiae) that requires agonist dependent M3 receptor activation for cell growth. By using receptor random mutagenesis followed by a genetic screen in yeast, we initially identified a point mutation at the cytoplasmic end of transmembrane domain (TM) VI (Q490L) that led to robust agonist-independent M3 receptor signaling in both yeast and mammalian cells. To explore further the molecular mechanisms by which point mutations can render GPCRs constitutively active, we subjected a region of the Q490L mutant M3 receptor that included TM V-VII to random mutagenesis. We then applied a yeast genetic screen to identify second-site mutations that could suppress the activating effects of the Q490L mutation and restore wild-type receptor-like function to the Q490L mutant receptor. This analysis led to the identification of 12 point mutations that allowed the Q490L mutant receptor to function in a fashion similar to the wild-type receptor. These amino acid substitutions mapped to two distinct regions of the M3 receptor, the exofacial segments of TM V and VI and the cytoplasmic ends of TM V-VII. Strikingly, in the absence of the activating Q490L mutation, all recovered point mutations severely reduced the efficiency of receptor/G protein coupling, indicating that the targeted residues play important roles in receptor activation and/or receptor/G protein coupling. This strategy should be generally applicable to identify sites in GPCRs that are critically involved in receptor function. PMID- 12750376 TI - A role for epsin N-terminal homology/AP180 N-terminal homology (ENTH/ANTH) domains in tubulin binding. AB - The epsin N-terminal homology (ENTH) domain is a protein module of approximately 150 amino acids found at the N terminus of a variety of proteins identified in yeast, plants, nematode, frog, and mammals. ENTH domains comprise multiple alpha helices folded upon each other to form a compact globular structure that has been implicated in interactions with lipids and proteins. In characterizing this evolutionarily conserved domain, we isolated and identified tubulin as an ENTH domain-binding partner. The interaction, which is direct and has a dissociation constant of approximately 1 microm, was observed with ENTH domains of proteins present in various species. Tubulin is co-immunoprecipitated from rat brain extracts with the ENTH domain-containing proteins, epsins 1 and 2, and punctate epsin staining is observed along the microtubule cytoskeleton of dissociated cortical neurons. Consistent with a role in microtubule processes, the over expression of epsin ENTH domain in PC12 cells stimulates neurite outgrowth. These data demonstrate an evolutionarily conserved property of ENTH domains to interact with tubulin and microtubules. PMID- 12750377 TI - Characterization of the final two genes of the gibberellin biosynthesis gene cluster of Gibberella fujikuroi: des and P450-3 encode GA4 desaturase and the 13 hydroxylase, respectively. AB - Recently, six genes of the gibberellin (GA) biosynthesis gene cluster in Gibberella fujikuroi were cloned and the functions of five of these genes were determined. Here we describe the function of the sixth gene, P450-3, and the cloning and functional analysis of a seventh gene, orf3, located at the left border of the gene cluster. We have thereby defined the complete GA biosynthesis gene cluster in this fungus. The predicted amino acid sequence of orf3 revealed no close homology to known proteins. High performance liquid chromatography and gas chromatography-mass spectrometry analyses of the culture fluid of knock-out mutants identified GA1 and GA4, rather than GA3 and GA7, as the major C19-GA products, suggesting that orf3 encodes the GA4 1,2-desaturase. This was confirmed by transformation of the SG139 mutant, which lacks the GA biosynthesis gene cluster, with the desaturase gene renamed des. The transformants converted GA4 to GA7, and also metabolized GA9 (3-deoxyGA4) to GA120 (1,2-didehydroGA9), but the 2alpha-hydroxylated compound GA40 was the major product in this case. We demonstrate also by gene disruption that P450-3, one of the four cytochrome P450 monooxygenase genes in the GA gene cluster, encodes the 13-hydroxylase, which catalyzes the conversion of GA7 to GA3, in the last step of the pathway. This enzyme also catalyzes the 13-hydroxylation of GA4 to GA1. Disruption of the des gene in an UV-induced P450-3 mutant produced a double mutant lacking both desaturase and 13-hydroxylase activities that accumulated high amounts of the commercially important GA4. The des and P450-3 genes differ in their regulation by nitrogen metabolite repression. In common with the other five GA biosynthesis genes, expression of the desaturase gene is repressed by high amounts of nitrogen in the culture medium, whereas P450-3 is the only gene in the cluster not repressed by nitrogen. PMID- 12750378 TI - CAIR-1/BAG-3 abrogates heat shock protein-70 chaperone complex-mediated protein degradation: accumulation of poly-ubiquitinated Hsp90 client proteins. AB - BAG family proteins are regulatory co-chaperones for heat shock protein (Hsp) 70. Hsp70 facilitates the removal of injured proteins by ubiquitin-mediated proteasomal degradation. This process can be driven by geldanamycin, an irreversible blocker of Hsp90. We hypothesize that CAIR-1/BAG-3 inhibits Hsp mediated proteasomal degradation. Human breast cancer cells were engineered to overexpress either full-length CAIR-1 (FL), which binds Hsp70, or a BAG domain deletion mutant (dBAG) that cannot bind Hsp70. FL overexpression prevented geldanamycin-mediated loss of total and phospho-Akt and other Hsp client proteins. dBAG provided no protection, indicating a requirement for Hsp70 binding. Ubiquitinated Akt accumulated in FL-expressing cells, mimicking the effect of lactacystin proteasomal inhibition, indicating that CAIR-1 inhibits proteasomal degradation distal to protein ubiquitination in a BAG domain dependent manner. Protein protection in FL cells was generalizable to downstream Akt targets, GSK3beta, P70S6 kinase, CREB, and other Hsp client proteins, including Raf-1, cyclin-dependent kinase 4, and epidermal growth factor receptor. These findings suggest that Hsp70 is a chaperone driving a multiprotein degradation complex and that the inhibitory co-chaperone CAIR-1 functions distal to client ubiquitination. Furthermore, poly-ubiquitination is not sufficient for efficient proteasomal targeting of Hsp client proteins. PMID- 12750379 TI - An artificially designed pore-forming protein with anti-tumor effects. AB - Protein engineering is an emerging area that has expanded our understanding of protein folding and laid the groundwork for the creation of unprecedented structures with unique functions. We previously designed the first native-like pore-forming protein, small globular protein (SGP). We show here that this artificially engineered protein has membrane-disrupting properties and anti-tumor activity in several cancer animal models. We propose and validate a mechanism for the selectivity of SGP toward cell membranes in tumors. SGP is the prototype for a new class of artificial proteins designed for therapeutic applications. PMID- 12750380 TI - Mapping sigma 54-RNA polymerase interactions at the -24 consensus promoter element. AB - The sigma 54 promoter specificity factor is distinct from sigma 70-type factors. The sigma 54-RNA polymerase binds to promoters with conserved sequence elements at -24 and -12 and utilizes specialized enhancer-binding activators to convert, through an ATP-dependent process, closed promoter complexes to open promoter complexes. The interface between sigma 54-RNA polymerase and promoter DNA is poorly characterized, contrasting with sigma 70. Here, sigma 54 was modified with strategically positioned cleavage reagents to provide physical evidence that the highly conserved RpoN box motif of sigma 54 is close to and may therefore interact with the consensus -24 promoter element. We show that the spatial relationship between the sigma 54-RNA polymerase and the -24 promoter element remains unchanged during closed to open complex conversion and transcription initiation but changes during the early elongation phase. In contrast, the spatial relationship between sigma 54-RNA polymerase and the consensus -12 promoter element changes upon conversion of the closed promoter complex to an open one. We provide evidence that some -12 promoter region-sigma 54 interactions are dependent upon either the core RNA polymerase or a fork junction DNA structure at the -12-position, indicating that DNA fork junctions can substitute for core RNAP. We also show the beta-subunit flap domain contributes to different sets of sigma-promoter DNA interactions at sigma 54- and sigma 70-dependent promoters. PMID- 12750381 TI - Structure and ubiquitin binding of the ubiquitin-interacting motif. AB - Ubiquitylation is used to target proteins into a large number of different biological processes including proteasomal degradation, endocytosis, virus budding, and vacuolar protein sorting (Vps). Ubiquitylated proteins are typically recognized using one of several different conserved ubiquitin binding modules. Here, we report the crystal structure and ubiquitin binding properties of one such module, the ubiquitin-interacting motif (UIM). We found that UIM peptides from several proteins involved in endocytosis and vacuolar protein sorting including Hrs, Vps27p, Stam1, and Eps15 bound specifically, but with modest affinity (Kd = 0.1-1 mm), to free ubiquitin. Full affinity ubiquitin binding required the presence of conserved acidic patches at the N and C terminus of the UIM, as well as highly conserved central alanine and serine residues. NMR chemical shift perturbation mapping experiments demonstrated that all of these UIM peptides bind to the I44 surface of ubiquitin. The 1.45 A resolution crystal structure of the second yeast Vps27p UIM (Vps27p-2) revealed that the ubiquitin interacting motif forms an amphipathic helix. Although Vps27p-2 is monomeric in solution, the motif unexpectedly crystallized as an antiparallel four-helix bundle, and the potential biological implications of UIM oligomerization are therefore discussed. PMID- 12750383 TI - WRN interacts physically and functionally with the recombination mediator protein RAD52. AB - Werner syndrome (WS) is a premature aging disorder that predisposes affected individuals to cancer development. The affected gene, WRN, encodes an RecQ homologue whose precise biological function remains elusive. Altered DNA recombination is a hallmark of WS cells suggesting that WRN plays an important role in these pathways. Here we report a novel physical and functional interaction between WRN and the homologous recombination mediator protein RAD52. Fluorescence resonance energy transfer (FRET) analyses show that WRN and RAD52 form a complex in vivo that co-localizes in foci associated with arrested replication forks. Biochemical studies demonstrate that RAD52 both inhibits and enhances WRN helicase activity in a DNA structure-dependent manner, whereas WRN increases the efficiency of RAD52-mediated strand annealing between non-duplex DNA and homologous sequences contained within a double-stranded plasmid. These results suggest that coordinated WRN and RAD52 activities are involved in replication fork rescue after DNA damage. PMID- 12750382 TI - Proexosite-1 on prothrombin is a factor Va-dependent recognition site for the prothrombinase complex. AB - Although the contribution of basic residues of exosite-1 to the catalytic function of thrombin has been studied extensively, their role in the specificity of prothrombin recognition by factor Xa in the prothrombinase complex (factor Xa, factor Va, phosphatidylcholine/phosphatidylserine vesicles, and Ca2+) has not been examined. In this study, we prepared several mutants of prethrombin-1 (prothrombin lacking Gla and Kringle-1 domains) in which basic residues of this site (Arg35, Lys36, Arg67, Lys70, Arg73, Arg75, and Arg77 in chymotrypsinogen numbering) were individually substituted with a Glu. Following expression in mammalian cells and purification to homogeneity, these mutants were characterized with respect to their ability to function as zymogens for both factor Xa and the prothrombinase complex. Factor Xa by itself exhibited similar catalytic activity toward both the wild type and mutant substrates; however, its activity in the prothrombinase complex toward most of mutants was severely impaired. Further kinetic studies in the presence of Tyr63-sulfated hirudin-(54-65) peptide suggested that although the peptide inhibits the prothrombinase activation of the wild type zymogen with a KD of 0.5-0.7 microm, it is ineffective in inhibiting the activation of mutant zymogens (KD = 2-30 microm). These results suggest that basic residues of proexosite-1 on prothrombin are factor Va-dependent recognition sites for factor Xa in the prothrombinase complex. PMID- 12750384 TI - Two-peaked synchronization in day/night expression rhythms of the fibrinogen gene cluster in the mouse liver. AB - Genes expressed with day/night rhythms in the mouse liver were searched for by microarray analysis using an in-house array harboring mouse liver cDNAs. The rhythmic expression with a single peak and trough level was confirmed by RNA blot analysis for 3beta-Hsd and Gabarapl1 genes exhibiting a peak in the light phase and Spot14, Hspa8, Hspa5, and Hsp84-1 genes showing a peak in the dark phase. On the other hand, mRNA levels for all of the three fibrinogen subunits, Aalpha, Bbeta and gamma, exhibited two peaks each in the light and dark phases in a synchronized manner. This two-peaked rhythmic pattern of fibrinogen genes as well as the single peak-trough pattern of other genes was diminished or almost completely lost in the liver of Clock mutant mice, suggesting that the two-peaked expression is also under the control of oscillation-generating genes. In constant darkness, the first peak of the expression rhythm of fibrinogen genes was almost intact, but the second peak disappeared. Therefore, although the first peak in the subjective day is a component of the innate circadian rhythm, the second peak seems to require light stimuli. Fasting in constant darkness caused shifts of time phases of the circadian rhythms. Protein levels of the fibrinogen subunits in whole blood also exhibited circadian rhythms. In the mouse and human loci of the fibrinogen gene cluster, a number of sequence elements resembling circadian transcription factor-binding sites were found. The fibrinogen gene locus provides a unique system for the study of two-peaked day/night rhythms of gene expression in a synchronized form. PMID- 12750385 TI - Oxidized low density lipoprotein inhibits macrophage apoptosis by blocking ceramide generation, thereby maintaining protein kinase B activation and Bcl-XL levels. AB - Macrophages play a central role in the development and progression of atherosclerotic lesions. It is well known that oxidized low density lipoprotein (ox-LDL) promotes the recruitment of monocytes (which differentiate to macrophages) into the intima. We reported recently that ox-LDL blocks apoptosis in bone marrow-derived macrophages deprived of macrophage colony-stimulating factor (M-CSF) by a mechanism involving protein kinase B (PKB) (Hundal, R., Salh, B., Schrader, J., Gomez-Munoz, A., Duronio, V., and Steinbrecher, U. (2001) J. Lipid Res. 42, 1483-1491). The aims of the present study were 1) to define the apoptotic pathway involved in the pro-survival effect of ox-LDL; 2) to determine which PKB target mediated this effect; and 3) to identify mechanisms responsible for PKB activation by ox-LDL. Apoptosis following M-CSF withdrawal was accompanied by activation of the caspase 9-caspase 3 cascade and cytochrome c release from mitochondria, but the caspase 8 pathway was unaffected. M-CSF withdrawal resulted in a marked and selective reduction in Bcl-XL protein and mRNA levels, and this decrease was prevented by ox-LDL. The ability of ox-LDL to preserve Bcl-XL levels was blocked by NFkappaB antagonists, thereby implicating IkappaB kinase as a key PKB target. M-CSF deprivation resulted in activation of acid sphingomyelinase and an increase in ceramide levels. Desipramine (a sphingomyelinase inhibitor) prevented the increase in ceramide and inhibited apoptosis after M-CSF deprivation. Ox-LDL completely blocked the increase in acid sphingomyelinase activity as well as the increase in ceramide after M-CSF deprivation. Pretreatment of macrophages with C2-ceramide reversed the effect of ox-LDL on PKB and macrophage survival. These results indicate that ox-LDL prevents apoptosis in M-CSF-deprived macrophages at least in part by inhibiting acid sphingomyelinase. This in turn prevents ceramide-induced down-regulation of PKB, the activity of which is required to maintain production of Bcl-XL. PMID- 12750386 TI - Dorfin localizes to Lewy bodies and ubiquitylates synphilin-1. AB - Parkinson's disease (PD) is a neurodegenerative disease characterized by loss of nigra dopaminergic neurons. Lewy bodies (LBs) are a characteristic neuronal inclusion in PD brains. In this study, we report that Dorfin, a RING finger-type ubiquityl ligase for mutant superoxide dismutase-1, was localized with ubiquitin in LBs. Recently, synphilin-1 was identified to associate with alpha-synuclein and to be a major component of LBs. We found that overexpression of synphilin-1 in cultured cells led to the formation of large juxtanuclear inclusions, but showed no cytotoxicity. Dorfin colocalized in these large inclusions with ubiquitin and proteasomal components. In contrast to full-length synphilin-1, overexpression of the central portion of synphilin-1, including ankyrin-like repeats, a coiled-coil domain, and an ATP/GTP-binding domain, predominantly led to the formation of small punctate aggregates scattered throughout the cytoplasm and showed cytotoxic effects. Dorfin and ubiquitin did not localize in these small aggregates. Overexpression of the N or C terminus of synphilin-1 did not lead to the formation of any aggregates. Dorfin physically bound and ubiquitylated synphilin-1 through its central portion, but did not ubiquitylate wild-type or mutant alpha-synuclein. These results suggest that the central domain of synphilin-1 has an important role in the formation of aggregates and cytotoxicity and that Dorfin may be involved in the pathogenic process of PD and LB formation by ubiquitylation of synphilin-1. PMID- 12750387 TI - The beta-subunit of the protein-conducting channel of the endoplasmic reticulum functions as the guanine nucleotide exchange factor for the beta-subunit of the signal recognition particle receptor. AB - Cotranslational protein transport to the endoplasmic reticulum is controlled by the concerted interaction of three GTPases: the SRP54 subunit of the signal recognition particle (SRP) and the alpha- and beta-subunits of the SRP receptor (SR). SRbeta is related to ADP-ribosylation factor (ARF)-type GTPases, and the recently published crystal structure of SRbeta-GTP in complex with the binding domain of SRalpha suggested that SRbeta, like all ARF-type GT-Pases, requires a guanine nucleotide exchange factor (GEF) for function. Searching the sequence data base, we identified significant sequence similarity between the Sec7 domain of ARF-GEFs and the cytosolic domains of the beta-subunits of the two homologous heterotrimeric protein-conducting channels in yeast. Using a fluorescence nucleotide exchange assay, we show that the beta-subunits of the heterotrimeric protein-conducting channels function as the GEFs for SRbeta. Both the cytosolic domain of Sec61beta as well as the holo-Sec61beta, when part of the isolated trimeric Sec61p complex, function as the GEF for SRbeta, whereas the same Sec61beta, when part of the heptameric complex that facilitates posttranslational protein transport, is inactive as the GEF for SRbeta PMID- 12750388 TI - Tumor suppressor p53 and its homologue p73alpha affect cell migration. AB - The p53 tumor suppressor plays a central role in the negative control of growth and survival of abnormal cells. Previously we demonstrated that in addition to these functions, p53 expression affects cell morphology and lamellar activity of the cell edge (Alexandrova, A., Ivanov, A., Chumakov, P. M., Kopnin, P. B., and Vasiliev, J. M. (2000) Oncogene 19, 5826-5830). In the present work we studied the effects of p53 and its homologue p73alpha on cell migration. We found that loss of p53 function correlated with decreased cell migration that was analyzed by in vitro wound closure test and Boyden chamber assay. The decreased motility of p53-deficient cells was observed in different cell contexts: human foreskin fibroblasts (BJ), human colon and lung carcinoma cell lines (HCT116 and H1299, respectively), as well as mouse normal fibroblasts from lung and spleen, peritoneal macrophages, and keratinocytes. On the other hand, overexpression of the p53 family member p73alpha stimulated cell migration. Changes in cell migration correlated directly with transcription activation induced by p53 or p73alpha. Noteworthy, p53 modulated cell motility in the absence of stress. The effect of p53 and p73alpha on cell migration was mediated through the activity of the phosphatidylinositol 3-kinase/Rac1 pathway. This p53/p73 function was mainly associated with some modulation of intracellular signaling rather than with stimulation of production of secreted motogenic factors. The identified novel activity of the p53 family members might be involved in regulation of embryogenesis, wound healing, or inflammatory response. PMID- 12750389 TI - Cytokine-induced patterns of gene expression in skeletal muscle tissue. AB - Tumor necrosis factor alpha (TNF-alpha) and other cytokines induce a state of negative energy balance leading to the breakdown of skeletal muscle. Leptin, another member of the cytokine superfamily, also induces a state of negative energy balance but does not alter lean body mass. The transcription profile of skeletal muscle was compared in animals treated with TNF-alpha or leptin or in animals pair-fed over a 7-day time course using 11,000-gene microarrays (Affymetrix, Santa Clara, CA). Ten clusters of skeletal muscle genes were identified, each of which showed significantly different expression between TNF alpha treatment and pair feeding. Studies comparing leptin treatment and pair feeding revealed that both activate nearly identical programs of gene expression in skeletal muscle. These data indicate that the effects of leptin on skeletal muscle are markedly different from those of TNF-alpha and that the effects of leptin on skeletal muscle can be largely ascribed to its anorectic effects. Subtle differences between leptin and pair feeding were evident only after 7 days of treatment. In general, pair feeding altered gene expression after the 7-day treatment, whereas leptin did not. The effects of TNF-alpha on skeletal muscle are distinct from those of pair feeding, a result consistent with its known catabolic effects on this tissue. Analyses of the data from food-restricted animals also identified a set of transcriptional changes associated with this state. Further studies of many newly identified leptin-, TNF-alpha-, and starvation-regulated genes and the apparent coordinate regulation of these clusters may reveal important insights into the different effects of cytokines on skeletal muscle. PMID- 12750390 TI - Imaging and biological function in health and disease. PMID- 12750391 TI - The addicted human brain: insights from imaging studies. PMID- 12750392 TI - Pulmonary surfactant: a front line of lung host defense. PMID- 12750393 TI - The mitochondriotoxic domain of Vpr determines HIV-1 virulence. PMID- 12750394 TI - A matter of life and death: cardiac myocyte apoptosis and regeneration. PMID- 12750395 TI - CD44--a sticky target for asthma. PMID- 12750396 TI - Activation of Mst1 causes dilated cardiomyopathy by stimulating apoptosis without compensatory ventricular myocyte hypertrophy. AB - Activation of mammalian sterile 20-like kinase 1 (Mst1) by genotoxic compounds is known to stimulate apoptosis in some cell types. The importance of Mst1 in cell death caused by clinically relevant pathologic stimuli is unknown, however. In this study, we show that Mst1 is a prominent myelin basic protein kinase activated by proapoptotic stimuli in cardiac myocytes and that Mst1 causes cardiac myocyte apoptosis in vitro in a kinase activity-dependent manner. In vivo, cardiac-specific overexpression of Mst1 in transgenic mice results in activation of caspases, increased apoptosis, and dilated cardiomyopathy. Surprisingly, however, Mst1 prevents compensatory cardiac myocyte elongation or hypertrophy despite increased wall stress, thereby obscuring the use of the Frank Starling mechanism, a fundamental mechanism by which the heart maintains cardiac output in response to increased mechanical load at the single myocyte level. Furthermore, Mst1 is activated by ischemia/reperfusion in the mouse heart in vivo. Suppression of endogenous Mst1 by cardiac-specific overexpression of dominant-negative Mst1 in transgenic mice prevents myocyte death by pathologic insults. These results show that Mst1 works as both an essential initiator of apoptosis and an inhibitor of hypertrophy in cardiac myocytes, resulting in a previously unrecognized form of cardiomyopathy. PMID- 12750397 TI - Targeted inhibition of p38 MAPK promotes hypertrophic cardiomyopathy through upregulation of calcineurin-NFAT signaling. AB - The MAPKs are important transducers of growth and stress stimuli in virtually all eukaryotic cell types. In the mammalian heart, MAPK signaling pathways have been hypothesized to regulate myocyte growth in response to developmental signals or physiologic and pathologic stimuli. Here we generated cardiac-specific transgenic mice expressing dominant-negative mutants of p38alpha, MKK3, or MKK6. Remarkably, attenuation of cardiac p38 activity produced a progressive growth response and myopathy in the heart that correlated with the degree of enzymatic inhibition. Moreover, dominant-negative p38alpha, MKK3, and MKK6 transgenic mice each showed enhanced cardiac hypertrophy following aortic banding, Ang II infusion, isoproterenol infusion, or phenylephrine infusion for 14 days. A mechanism underlying this enhanced-growth profile was suggested by the observation that dominant-negative p38alpha directly augmented nuclear factor of activated T cells (NFAT) transcriptional activity and its nuclear translocation. In vivo, NFAT dependent luciferase reporter transgenic mice showed enhanced activation in the presence of the dominant-negative p38alpha transgene before and after the onset of cardiac hypertrophy. More significantly, genetic disruption of the calcineurin Abeta gene rescued hypertrophic cardiomyopathy and depressed functional capacity observed in p38-inhibited mice. Collectively, these observations indicate that reduced p38 signaling in the heart promotes myocyte growth through a mechanism involving enhanced calcineurin-NFAT signaling. PMID- 12750398 TI - Antigenic drift as a mechanism for tumor evasion of destruction by cytolytic T lymphocytes. AB - It is established that mutations in viral antigenic epitopes, or antigenic drifts, allow viruses to escape recognition by both Ab's and T lymphocytes. It is unclear, however, whether tumor cells can escape immune recognition via antigenic drift. Here we show that adoptive therapy with both monoclonal and polyclonal transgenic CTLs, specific for a natural tumor antigen, P1A, selects for multiple mutations in the P1A antigenic epitope. These mutations severely diminish T cell recognition of the tumor antigen by a variety of mechanisms, including modulation of MHC:peptide interaction and TCR binding to MHC:peptide complex. These results provide the first evidence for tumor evasion of T cell recognition by antigenic drift, and thus have important implications for the strategy of tumor immunotherapy. PMID- 12750400 TI - CD137 costimulatory T cell receptor engagement reverses acute disease in lupus prone NZB x NZW F1 mice. AB - Systemic lupus erythematosus (SLE) is a CD4(+) T cell-dependent, immune complex mediated, autoimmune disease that primarily affects women of childbearing age. Generation of high-titer affinity-matured IgG autoantibodies, specific for double stranded DNA and other nuclear antigens, coincides with disease progression. Current forms of treatment of SLE including glucocorticosteroids are often inadequate and induce severe side effects. Immunological approaches for treating SLE in mice using anti-CD4 mAb's or CTLA4-Ig and anti-CD154 mAb's have proven to be effective. However, like steroid treatment, these regimens induce global immunosuppression, and their withdrawal allows for disease progression. In this report we show that lupus-prone NZB x NZW F(1) mice given three injections of anti-CD137 (4-1BB) mAb's between 26 and 35 weeks of age reversed acute disease, blocked chronic disease, and extended the mice's lifespan from 10 months to more than 2 years. Autoantibody production in recipients was rapidly suppressed without inducing immunosuppression. Successful treatment could be traced to the fact that NZB x NZW F(1) mice, regardless of their age or disease status, could not maintain pathogenic IgG autoantibody production in the absence of continuous CD4(+) T cell help. Our data support the hypothesis that CD137-mediated signaling anergized CD4(+) T cells during priming at the DC interface. PMID- 12750399 TI - A mechanistic role for cardiac myocyte apoptosis in heart failure. AB - Heart failure is a common, lethal condition whose pathogenesis is poorly understood. Recent studies have identified low levels of myocyte apoptosis (80 250 myocytes per 10(5) nuclei) in failing human hearts. It remains unclear, however, whether this cell death is a coincidental finding, a protective process, or a causal component in pathogenesis. Using transgenic mice that express a conditionally active caspase exclusively in the myocardium, we demonstrate that very low levels of myocyte apoptosis (23 myocytes per 10(5) nuclei, compared with 1.5 myocytes per 10(5) nuclei in controls) are sufficient to cause a lethal, dilated cardiomyopathy. Interestingly, these levels are four- to tenfold lower than those observed in failing human hearts. Conversely, inhibition of cardiac myocyte death in this murine model largely prevents the development of cardiac dilation and contractile dysfunction, the hallmarks of heart failure. To our knowledge, these data provide the first direct evidence that myocyte apoptosis may be a causal mechanism of heart failure, and they suggest that inhibition of this cell death process may constitute the basis for novel therapies. PMID- 12750402 TI - Functional role of inward rectifier current in heart probed by Kir2.1 overexpression and dominant-negative suppression. AB - The inward rectifier current I(K1) is tightly regulated regionally within the heart, downregulated in heart failure, and genetically suppressed in Andersen syndrome. We used in vivo viral gene transfer to dissect the role of I(K1) in cardiac repolarization and maintenance of the resting membrane potential (RMP) in guinea pig ventricular myocytes. Kir2.1 overexpression boosted Ba(2+)-sensitive I(K1) by more than 100% (at -50mV), significantly shortened action potential durations (APDs), accelerated phase 3 repolarization, and hyperpolarized RMP compared with control cells (nongreen cells from the same hearts and green cells from GFP-transduced hearts). The dominant-negative Kir2.1AAA reduced I(K1) by 50 90%; those cells with less than 80% reduction of I(K1) exhibited prolonged APDs, decelerated phase 3 repolarization, and depolarization of the RMP. Further reduction of I(K1) resulted in a pacemaker phenotype, as previously described. ECGs revealed a 7.7% +/- 0.9% shortening of the heart rate-corrected QT interval (QTc interval) in Kir2.1-transduced animals (n = 4) and a 16.7% +/- 1.8% prolongation of the QTc interval (n = 3) in Kir2.1AAA-transduced animals 72 hours after gene delivery compared with immediate postoperative recordings. Thus, I(K1) is essential for establishing the distinctive electrical phenotype of the ventricular myocyte: rapid terminal repolarization to a stable and polarized resting potential. Additionally, the long-QT phenotype seen in Andersen syndrome is a direct consequence of dominant-negative suppression of Kir2 channel function. PMID- 12750401 TI - Heterozygous deficiency of hypoxia-inducible factor-2alpha protects mice against pulmonary hypertension and right ventricular dysfunction during prolonged hypoxia. AB - Chronic hypoxia induces pulmonary vascular remodeling, leading to pulmonary hypertension, right ventricular hypertrophy, and heart failure. Heterozygous deficiency of hypoxia-inducible factor-1alpha (HIF-1alpha), which mediates the cellular response to hypoxia by increasing expression of genes involved in erythropoiesis and angiogenesis, has been previously shown to delay hypoxia induced pulmonary hypertension. HIF-2alpha is a homologue of HIF-1alpha and is abundantly expressed in the lung, but its role in pulmonary hypertension remains unknown. Therefore, we analyzed the pulmonary response of WT and viable heterozygous HIF-2alpha-deficient (Hif2alpha(+/-)) mice after exposure to 10% O(2) for 4 weeks. In contrast to WT mice, Hif2alpha(+/-) mice were fully protected against pulmonary hypertension and right ventricular hypertrophy, unveiling a critical role of HIF-2alpha in hypoxia-induced pulmonary vascular remodeling. Pulmonary expression levels of endothelin-1 and plasma catecholamine levels were increased threefold and 12-fold respectively in WT but not in Hif2alpha(+/-) mice after hypoxia, suggesting that HIF-2alpha-mediated upregulation of these vasoconstrictors contributes to the development of hypoxic pulmonary vascular remodeling. PMID- 12750403 TI - Pacemaker channel dysfunction in a patient with sinus node disease. AB - The cardiac pacemaker current I(f) is a major determinant of diastolic depolarization in sinus nodal cells and has a key role in heartbeat generation. Therefore, we hypothesized that some forms of "idiopathic" sinus node dysfunction (SND) are related to inherited dysfunctions of cardiac pacemaker ion channels. In a candidate gene approach, a heterozygous 1-bp deletion (1631delC) in exon 5 of the human HCN4 gene was detected in a patient with idiopathic SND. The mutant HCN4 protein (HCN4-573X) had a truncated C-terminus and lacked the cyclic nucleotide-binding domain. COS-7 cells transiently transfected with HCN4-573X cDNA indicated normal intracellular trafficking and membrane integration of HCN4 573X subunits. Patch-clamp experiments showed that HCN4-573X channels mediated I(f)-like currents that were insensitive to increased cellular cAMP levels. Coexpression experiments showed a dominant-negative effect of HCN4-573X subunits on wild-type subunits. These data indicate that the cardiac I(f) channels are functionally expressed but with altered biophysical properties. Taken together, the clinical, genetic, and in vitro data provide a likely explanation for the patient's sinus bradycardia and the chronotropic incompetence. PMID- 12750404 TI - Vpr R77Q is associated with long-term nonprogressive HIV infection and impaired induction of apoptosis. AB - The absence of immune defects that occurs in the syndrome of long-term nonprogressive (LTNP) HIV infection offers insights into the pathophysiology of HIV-induced immune disease. The (H[F/S]RIG)(2) domain of viral protein R (Vpr) induces apoptosis and may contribute to HIV-induced T cell depletion. We demonstrate a higher frequency of R77Q Vpr mutations in patients with LTNP than in patients with progressive disease. In addition, T cell infections using vesicular stomatitis virus G (VSV-G) pseudotyped HIV-1 Vpr R77Q result in less (P = 0.01) T cell death than infections using wild-type Vpr, despite similar levels of viral replication. Wild-type Vpr-associated events, including procaspase-8 and -3 cleavage, loss of mitochondrial transmembrane potential (deltapsi(m)), and DNA fragmentation factor activation are attenuated by R77Q Vpr. These data highlight the pathophysiologic role of Vpr in HIV-induced immune disease and suggest a novel mechanism of LTNP. PMID- 12750405 TI - GLUT4, AMP kinase, but not the insulin receptor, are required for hepatoportal glucose sensor-stimulated muscle glucose utilization. AB - Recent evidence suggests the existence of a hepatoportal vein glucose sensor, whose activation leads to enhanced glucose use in skeletal muscle, heart, and brown adipose tissue. The mechanism leading to this increase in whole body glucose clearance is not known, but previous data suggest that it is insulin independent. Here, we sought to further determine the portal sensor signaling pathway by selectively evaluating its dependence on muscle GLUT4, insulin receptor, and the evolutionarily conserved sensor of metabolic stress, AMP activated protein kinase (AMPK). We demonstrate that the increase in muscle glucose use was suppressed in mice lacking the expression of GLUT4 in the organ muscle. In contrast, glucose use was stimulated normally in mice with muscle specific inactivation of the insulin receptor gene, confirming independence from insulin-signaling pathways. Most importantly, the muscle glucose use in response to activation of the hepatoportal vein glucose sensor was completely dependent on the activity of AMPK, because enhanced hexose disposal was prevented by expression of a dominant negative AMPK in muscle. These data demonstrate that the portal sensor induces glucose use and development of hypoglycemia independently of insulin action, but by a mechanism that requires activation of the AMPK and the presence of GLUT4. PMID- 12750406 TI - A role for CD44 in an antigen-induced murine model of pulmonary eosinophilia. AB - Previous studies established that IL-5-producing CD4(+) T cells play a pivotal role in allergic respiratory inflammation. It was also reported that CD4(+) T cells express higher levels of CD44 in the airway than in peripheral blood of patients with allergic respiratory diseases. We have used experimental pulmonary eosinophilia induced in mice by Ascaris suum (Asc) extract to investigate the role of CD44 in the development of allergic respiratory inflammation. Intraperitoneal administration of anti-CD44 mAb prevented both lymphocyte and eosinophil accumulation in the lung. Anti-CD44 mAb also blocked antigen-induced elevation of Th2 cytokines as well as chemokines (CCL11, CCL17) in bronchoalveolar lavage fluid (BALF). Treatment with anti-CD44 mAb inhibited the increased levels of hyaluronic acid (HA) and leukotriene concentrations in BALF that typically result from antigen challenge. Anti-CD44 mAb also blocked antigen induced airway hyperresponsiveness. An anti-CD44 mAb (IM7) inhibited the HA binding ability of splenocytes associated with decreased levels of CD44. Soluble CD44 levels in serum were increased in Asc-challenged IM7-treated mice, but not in KM201-treated mice, compared with Asc-challenged rat IgG-treated mice. Ab's that block CD44-HA binding reduced allergic respiratory inflammation by preventing lymphocyte and eosinophil accumulation in the lung. Thus, CD44 may be critical for development of allergic respiratory inflammation. PMID- 12750407 TI - Critical role of the Toll-like receptor signal adaptor protein MyD88 in acute allograft rejection. AB - The Toll-like receptors (TLRs) are recently discovered germline-encoded receptors on APCs that are critically important in innate immune recognition of microbial pathogens. However, their role in solid-organ transplantation is unknown. To explore this role, we employed a skin allograft model using mice with targeted deletion of the universal TLR signal adaptor protein, MyD88. We report that minor antigen-mismatched (HY-mismatched) allograft rejection cannot occur in the absence of MyD88 signaling. Furthermore, we show that the inability to reject these allografts results from a reduced number of mature DCs in draining lymph nodes, leading to impaired generation of anti-graft-reactive T cells and impaired Th1 immunity. Hence, this work demonstrates that TLRs can be activated in a transplant setting and not solely by infections. These results link innate immunity to the initiation of the adaptive alloimmune response. PMID- 12750408 TI - HDL-associated estradiol stimulates endothelial NO synthase and vasodilation in an SR-BI-dependent manner. AB - Cardiovascular diseases remain the leading cause of death in the United States. Two factors associated with a decreased risk of developing cardiovascular disease are elevated HDL levels and sex - specifically, a decreased risk is found in premenopausal women. HDL and estrogen stimulate eNOS and the production of nitric oxide, which has numerous protective effects in the vascular system including vasodilation, antiadhesion, and anti-inflammatory effects. We tested the hypothesis that HDL binds to its receptor, scavenger receptor class B type I (SR BI), and delivers estrogen to eNOS, thereby stimulating the enzyme. HDL isolated from women stimulated eNOS, whereas HDL isolated from men had minimal activity. Studies with ovariectomized and ovariectomized/estrogen replacement mouse models demonstrated that HDL-associated estradiol stimulation of eNOS is SR-BI dependent. Furthermore, female HDL, but not male HDL, promoted the relaxation of muscle strips isolated from C57BL/6 mice but not SR-BI null mice. Finally, HDL isolated from premenopausal women or postmenopausal women receiving estradiol replacement therapy stimulated eNOS, whereas HDL isolated from postmenopausal women did not stimulate eNOS. We conclude that HDL-associated estrodial is capable of the stimulating eNOS. These studies establish a new paradigm for examining the cardiovascular effects of HDL and estrogen. PMID- 12750410 TI - Band-pass response properties of rat SI neurons. AB - Rats typically employ 4- to 12-Hz "whisking" movements of their vibrissae during tactile exploration. The intentional sampling of signals in this frequency range suggests that neural processing of tactile information may be differentially engaged in this bandwidth. We examined action potential responses in rat primary somatosensory cortex (SI) to a range of frequencies of vibrissa motion. Single vibrissae were mechanically deflected with 5-s pulse trains at rates /=3 Hz. In contrast with this low-pass feature of the response, several other characteristics of the response revealed bandpass response properties. While average evoked response amplitudes measured 0 35 ms after stimulus onset typically decreased with increasing frequency, the later components of the response (>15 ms post stimulus) were augmented at frequencies between 3 and 10 Hz. Further, during the steady state, both rate and temporal measures of neural activity, measured as total spike rate or vector strength (a measure of temporal fidelity of spike timing across cycles), showed peak signal values at 5-10 Hz. A minimal biophysical network model of SI layer IV, consisting of an excitatory and inhibitory neuron and thalamocortical input, captured the spike rate and vector strength band-pass characteristics. Further analyses in which specific elements were selectively removed from the model suggest that slow inhibitory influences give rise to the band-pass peak in temporal precision, while thalamocortical adaptation can account for the band pass peak in spike rate. The presence of these band-pass characteristics may be a general property of thalamocortical circuits that lead rodents to target this frequency range with their whisking behavior. PMID- 12750411 TI - Mu-opioid receptors facilitate the propagation of excitatory activity in rat hippocampal area CA1 by disinhibition of all anatomical layers. AB - Hippocampal mu-opioid receptors (MORs) have been implicated in memory formation associated with opiate drug abuse. MORs modulate hippocampal synaptic plasticity acutely, when chronically activated, and during drug withdrawal. At the network level, MORs increase excitability in area CA1 by disinhibiting pyramidal cells. The precise inhibitory interneuron subtypes affected by MOR activation are unknown; however, not all subtypes are inhibited, and specific interneuron subtypes have been shown to preferentially express MORs. Here we investigate, using voltage-sensitive dye imaging in brain slices, the effect of MOR activation on the patterns of inhibition and on the propagation of excitatory activity in rat hippocampal CA1. MOR activation augments excitatory activity evoked by stimulating inputs in stratum oriens [i.e., Schaffer collateral and commissural pathway (SCC) and antidromic], stratum radiatum (i.e., SCC), and stratum lacunosum-moleculare (SLM; i.e., perforant path and thalamus). The augmented excitatory activity is further facilitated as it propagates through the CA1 network. This was observed as a proportionately larger increase in amplitudes of excitatory activity at sites distal from where the activity was evoked. This facilitation was observed for excitatory activity propagating from all three stimulation sites. The augmentation and facilitation were prevented by GABAA receptor antagonists (bicuculline, 30 microM), but not by GABAB receptor antagonists (CGP 55845, 10 microM). Furthermore, MOR activation inhibited IPSPs in all layers of area CA1. These findings suggest that MOR-induced suppression of GABA release onto GABAA receptors augments all inputs to CA1 pyramidal cells and facilitates the propagation of excitatory activity through the network of area CA1. PMID- 12750409 TI - Surfactant proteins A and D inhibit the growth of Gram-negative bacteria by increasing membrane permeability. AB - The pulmonary collectins, surfactant proteins A (SP-A) and D (SP-D), have been reported to bind lipopolysaccharide (LPS), opsonize microorganisms, and enhance the clearance of lung pathogens. In this study, we examined the effect of SP-A and SP-D on the growth and viability of Gram-negative bacteria. The pulmonary clearance of Escherichia coli K12 was reduced in SP-A-null mice and was increased in SP-D-overexpressing mice, compared with strain-matched wild-type controls. Purified SP-A and SP-D inhibited bacterial synthetic functions of several, but not all, strains of E. coli, Klebsiella pneumoniae, and Enterobacter aerogenes. In general, rough E. coli strains were more susceptible than smooth strains, and collectin-mediated growth inhibition was partially blocked by coincubation with rough LPS vesicles. Although both SP-A and SP-D agglutinated E. coli K12 in a calcium-dependent manner, microbial growth inhibition was independent of bacterial aggregation. At least part of the antimicrobial activity of SP-A and SP D was localized to their C-terminal domains using truncated recombinant proteins. Incubation of E. coli K12 with SP-A or SP-D increased bacterial permeability. Deletion of the E. coli OmpA gene from a collectin-resistant smooth E. coli strain enhanced SP-A and SP-D-mediated growth inhibition. These data indicate that SP-A and SP-D are antimicrobial proteins that directly inhibit the proliferation of Gram-negative bacteria in a macrophage- and aggregation independent manner by increasing the permeability of the microbial cell membrane. PMID- 12750412 TI - Activity profiles of single neurons in caudal anterior cingulate cortex during trace eyeblink conditioning in the rabbit. AB - Acquisition of trace eyeblink conditioning involves the association of a conditioned stimulus (CS) with an unconditioned stimulus (US) separated by a stimulus-free trace interval. This form of conditioning is dependent upon the hippocampus and the caudal anterior cingulate cortex (AC), in addition to brain stem and cerebellar circuitry. Hippocampal involvement in trace eyeblink conditioning has been studied extensively, but the involvement of caudal AC is less well understood. In the present study, we compared neuronal responses from rabbits given either paired (trace conditioning) or unpaired (pseudoconditioning) presentations of the CS and US. Presentation of the CS elicited significant increases in neuronal activity at the onset of both trace conditioning and pseudoconditioning. A robust CS-elicited neuronal response persisted throughout the first 2 days of trace conditioning, declining gradually across subsequent training sessions. In contrast, the magnitude of the CS-elicited excitatory response during pseudoconditioning began to decline within the first 10 trials. Neurons exhibiting excitatory responses to the CS during trace conditioning also exhibited excitatory responses to the US that were significantly greater in magnitude than US-elicited responses during pseudoconditioning. CS-elicited decreases in neuronal activity became more robust over the course of trace conditioning compared to pseudoconditioning. Reductions in activity during the CS interval consistently preceded excitation in both training groups, suggesting that the CS-elicited decreases in neuronal activity may serve to increase the signal-to-noise ratio of the excitatory response to the tone. Taken together, these data suggest that the caudal AC is involved early in trace eyeblink conditioning and that maintenance of the CS-elicited excitatory response may serve to signal the salience of the tone. PMID- 12750414 TI - A functional-anatomical model for lipreading. AB - Regional cerebral blood flow (rCBF) PET scans were used to study the physiological bases of lipreading, a natural skill of extracting language from mouth movements, which contributes to speech perception in everyday life. Viewing connected mouth movements that could not be lexically identified and that evoke perception of isolated speech sounds (nonlexical lipreading) was associated with bilateral activation of the auditory association cortex around Wernicke's area, of left dorsal premotor cortex, and left opercular-premotor division of the left inferior frontal gyrus (Broca's area). The supplementary motor area was active as well. These areas have all been implicated in phonological processing, speech and mouth motor planning, and execution. In addition, nonlexical lipreading also differentially activated visual motion areas. Lexical access through lipreading was associated with a similar pattern of activation and with additional foci in ventral- and dorsolateral prefrontal cortex bilaterally and in left inferior parietal cortex. Linear regression analysis of cerebral blood flow and proficiency for lexical lipreading further clarified the role of these areas in gaining access to language through lipreading. The results suggest cortical activation circuits for lipreading from action representations that may differentiate lexical access from nonlexical processes. PMID- 12750413 TI - GABA receptor-mediated inhibition of neuronal activity in rat SCN in vitro: pharmacology and influence of circadian phase. AB - The effect of gamma-aminobutyric acid (GABA) on neuronal firing rate in rat suprachiasmatic nucleus (SCN) slices was examined using continuous recording methods. GABA inhibited neuronal discharge during both the subjective day and the subjective night in a concentration-dependent manner characterized by two apparent affinity states. The GABAA receptor agonist muscimol caused potent inhibition regardless of circadian time; repeated applications of the agonist did not reverse the direction of effect. The GABAA receptor antagonists bicuculline and picrotoxin increased excitability when applied during either subjective day or subjective night. A significant increase in GABAA receptor- mediated inhibition, as well as endogenous GABAergic tone, was observed on the second day after slice preparation. The GABAB receptor agonist baclofen inhibited cell firing during subjective day and night, but the GABAB antagonist phaclofen had no significant effect. These data provide additional strong support for a predominantly inhibitory role of GABA in the rat SCN, regardless of the time of application in relation to the circadian rhythm, and demonstrate an important level of plasticity of this system in vitro. PMID- 12750415 TI - Temporal frequency and velocity-like tuning in the pigeon accessory optic system. AB - Neurons in the accessory optic system (AOS) and pretectum are involved in the analysis of optic flow and the generation of the optokinetic response. Previous studies found that neurons in the pretectum and AOS exhibit direction selectivity in response to large-field motion and are tuned in the spatiotemporal domain. Furthermore, it has been emphasized that pretectal and AOS neurons are tuned to a particular temporal frequency, consistent with the "correlation" model of motion detection. We examined the responses of neurons in the nucleus of the basal optic root (nBOR) of the AOS in pigeons to large-field drifting sine wave gratings of varying spatial (SF) and temporal frequencies (TF). nBOR neurons clustered into two categories: "Fast" neurons preferred low SFs and high TFs, and "Slow" neurons preferred high SFs and low TFs. The fast neurons were tuned for TF, but the slow nBOR neurons had spatiotemporally oriented peaks that suggested velocity tuning (TF/SF). However, the peak response was not independent of SF; thus we refer to the tuning as "apparent velocity tuning" or "velocity-like tuning." Some neurons showed peaks in both the fast and slow regions. These neurons were TF-tuned at low SFs, and showed velocity-like tuning at high SFs. We used computer simulations of the response of an elaborated Reichardt detector to show that both the TF-tuning and velocity-like tuning shown by the fast and slow neurons, respectively, may be explained by modified versions of the correlation model of motion detection. PMID- 12750416 TI - MSTd neuronal basis functions for the population encoding of heading direction. AB - Basis functions have been extensively used in models of neural computation because they can be combined linearly to approximate any nonlinear functions of the encoded variables. We investigated whether dorsal medial superior temporal (MSTd) area neurons use basis functions to simultaneously encode heading direction, eye position, and the velocity of ocular pursuit. Using optimal linear estimators, we first show that the head-centered and eye-centered position of a focus of expansion (FOE) in optic flow, pursuit direction, and eye position can all be estimated from the single-trial responses of 144 MSTd neurons with an average accuracy of 2-3 degrees, a value consistent with the discrimination thresholds measured in humans and monkeys. We then examined the format of the neural code for the head-centered position of the FOE, eye position, and pursuit direction. The basis function hypothesis predicts that a large majority of cells in MSTd should encode two or more signals simultaneously and combine these signals nonlinearly. Our analysis shows that 95% of the neurons encode two or more signals, whereas 76% code all three signals. Of the 95% of cells encoding two or more signals, 90% show nonlinear interactions between the encoded variables. These findings support the notion that MSTd may use basis functions to represent the FOE in optic flow, eye position, and pursuit. PMID- 12750417 TI - Synchrony levels during evoked seizure-like bursts in mouse neocortical slices. AB - Slices (n = 45) from the somatosensory cortex of mouse (P8-13) generated spontaneous bursts of activity (0.10 +/- 0.05 Hz) that were recorded extracellularly. Multiunit action potential (AP) activity was integrated and used as an index of population activity. In this experimental model, seizure-like activity (SLA) was evoked with bicuculline (5-10 microM) or N-methyl-d-aspartate (NMDA, 5 microM). SLA was an episode with repetitive bursting at a frequency of 0.50 +/- 0.06 Hz. To evaluate whether SLA was associated with a change in synchrony, we obtained simultaneous intracellular and extracellular recordings (n = 40) and quantified the relationship between individual cells and the surrounding population of neurons. During the SLA there was an increase in population activity and bursting activity was observed in neurons and areas that were previously silent. We defined synchrony as cellular activity that is consistently locked with the population bursts. Signal-averaging techniques were used to determine this component. To quantitatively assess change in synchronous activity at SLA onset, we estimated the entropy of the single cell's spike trains and subdivided this measure into network burst-related information and noise related entropy. The burst-related information was not significantly altered at the onset of NMDA-evoked SLA and slightly increased when evoked with bicuculline. The signal-to-noise ratio determined from the entropy estimates showed a significant decrease (instead of an expected increase) during SLA. We conclude that the increased population activity during the SLA is attributed to recruitment of neurons rather than to increased synchrony of each of the individual elements. PMID- 12750418 TI - ERK integrates PKA and PKC signaling in superficial dorsal horn neurons. II. Modulation of neuronal excitability. AB - Protein kinases belonging to the protein kinase A (PKA), protein kinase C (PKC), and extracellular signal-related kinase (ERK) families have been identified as key players in modulating nociception at the level of the spinal cord dorsal horn, yet little is known about the effects of these kinases on membrane properties of the dorsal horn neurons. PKA, PKC, and ERK exert inhibitory effects on transient potassium currents (A-type currents or IA) in mouse superficial dorsal horn neurons (Hu et al. 2003). Here we aimed to determine the effects of these kinases on action potential firing and membrane properties of these neurons to evaluate the impact of the modulation of IA (and other conductances) in these neurons. We found that activating PKC and PKA has dramatic effects on action potential firing, reflecting an increase in the excitability of superficial dorsal horn neurons. In addition, we found that inhibitors of both PKC and ERK signaling decrease the excitability of dorsal horn neurons, suggesting that these kinases exert a tonic excitation of these cells. Consistent with our findings that these kinases inhibit A-type currents, we found that PKA, PKC, and ERK act to shorten the first-spike latency after depolarization induced by current injection. In addition, activation of these kinases increases spike frequency and action potential amplitude of dorsal horn neurons. Interestingly, we found that the effects of PKA and PKC activators are blocked by inhibitors of ERK signaling, suggesting that PKA and PKC may exert their actions by activation of ERKs. PMID- 12750419 TI - ERK integrates PKA and PKC signaling in superficial dorsal horn neurons. I. Modulation of A-type K+ currents. AB - The transient outward potassium currents (also known as A-type currents or IA) are important determinants of neuronal excitability. In the brain, IA is modulated by protein kinase C (PKC), protein kinase A (PKA), and extracellular signal-related kinase (ERK), three kinases that have been shown to be critical modulators of nociception. We wanted to determine the effects of these kinases on IA in superficial dorsal horn neurons. Using whole cell recordings from cultured mouse spinal cord superficial dorsal horn neurons, we found that PKC and PKA both inhibit IA in these cells, and that PKC has a tonic inhibitory action on IA. Further, we provide evidence supporting the hypothesis that PKC and PKA do not modulate IA directly, but rather act as upstream activators of ERKs, which modulate IA. These results suggest that ERKs serve as signal integrators in modulation of IA in dorsal horn neurons and that modulation of A-type potassium currents may underlie aspects of central sensitization mediated by PKC, PKA, and ERKs. PMID- 12750420 TI - Characterization of cutaneous primary afferent fibers excited by acetic acid in a model of nociception in frogs. AB - Acetic acid applied to the hind limb of a frog evokes nocifensive behaviors, including a vigorous wiping of the exposed skin, referred to as the wiping response. The aim of this study was to examine the responses of cutaneous primary afferent fibers in frogs to acetic acid (pH 2.84-1.42) applied topically to the skin. Conventional electrophysiological methods were used to record neuronal activity from single identified primary afferent fibers with cutaneous receptive fields on the hind limb. Fibers were classified according to their conduction velocities and responses evoked by mechanical and thermal (heat and cold) stimuli. One hundred and twenty-two mechanosensitive afferent fibers were studied (44 Abeta, 60 Adelta, and 18 C fibers). Thirty-nine percent of all fibers were excited by acetic acid, but a greater percentage of Adelta (52%) and C fibers (44%) were excited than Abeta fibers (20%). Evoked responses of fibers increased with increasingly more acidic pH until the greatest responses were evoked by acetic acid at pH 2.59-2.41. Application of acetic acid at pHs <2.41 evoked less excitation, suggesting that fibers became desensitized. Similar percentages of nociceptors and low-threshold mechanoreceptors were excited by acetic acid. Thus primary afferent fibers were excited by acetic acid at pHs that have been shown to evoke the wiping response in our previous study. The results of the present study suggest that the model of acetic acid-induced nociception in frogs may be useful for studying the mechanisms by which tissue acidosis produces pain. PMID- 12750421 TI - Voltage-gated Na+ channels enhance the temporal filtering properties of electrosensory neurons in the torus. AB - Regenerative processes enhance postsynaptic potential (PSP) amplitude and behaviorally relevant temporal filtering in more than one-third of electrosensory neurons in the torus semicircularis of Eigenmannia. Data from in vivo current clamp intracellular recordings indicate that these "regenerative PSPs" can be divided in two groups based on their half-amplitude durations: constant duration (CD) and variable duration (VD) PSPs. CD PSPs have half-amplitude durations of between 20 and 60 ms that do not vary in relation to stimulus periodicity. In contrast, the half-amplitude durations of VD PSPs vary in relation to stimulus periodicity and range from approximately 10 to 500 ms. Injection of 0.1 nA sinusoidal current through the recording electrode demonstrated that CD PSPs and not VD PSPs can be elicited by voltage fluctuations alone. In addition, CD PSPs were blocked by intracellular application of either QX-314 or QX-222, whereas VD PSPs were not. These in vivo data suggest, therefore, that CD PSPs are mediated by voltage-dependent Na+ conductances. PMID- 12750422 TI - Neocortical pyramidal cells respond as integrate-and-fire neurons to in vivo-like input currents. AB - In the intact brain neurons are constantly exposed to intense synaptic activity. This heavy barrage of excitatory and inhibitory inputs was recreated in vitro by injecting a noisy current, generated as an Ornstein-Uhlenbeck process, into the soma of rat neocortical pyramidal cells. The response to such in vivo-like currents was studied systematically by analyzing the time development of the instantaneous spike frequency, and when possible, the stationary mean spike frequency as a function of both the mean and the variance of the input current. All cells responded with an in vivo-like action potential activity with stationary statistics that could be sustained throughout long stimulation intervals (tens of seconds), provided the frequencies were not too high. The temporal evolution of the response revealed the presence of mechanisms of fast and slow spike frequency adaptation, and a medium duration mechanism of facilitation. For strong input currents, the slow adaptation mechanism made the spike frequency response nonstationary. The minimal frequencies that caused strong slow adaptation (a decrease in the spike rate by more than 1 Hz/s), were in the range 30-80 Hz and depended on the pipette solution used. The stationary response function has been fitted by two simple models of integrate-and-fire neurons endowed with a frequency-dependent modification of the input current. This accounts for all the fast and slow mechanisms of adaptation and facilitation that determine the stationary response, and proved necessary to fit the model to the experimental data. The coefficient of variability of the interspike interval was also in part captured by the model neurons, by tuning the parameters of the model to match the mean spike frequencies only. We conclude that the integrate and-fire model with spike-frequency-dependent adaptation/facilitation is an adequate model reduction of cortical cells when the mean spike-frequency response to in vivo-like currents with stationary statistics is considered. PMID- 12750423 TI - Responses to natural scenes in cat V1. AB - Studies on processing in primary visual areas often use artificial stimuli such as bars or gratings. As a result, little is known about the properties of activity patterns for the natural stimuli processed by the visual system on a daily basis. Furthermore, in the cat, a well-studied model system for visual processing, most results are obtained from anesthetized subjects and little is known about neuronal activations in the alert animal. Addressing these issues, we measure local field potentials (lfp) and multiunit spikes in the primary visual cortex of awake cats. We compare changes in the lfp power spectra and multiunit firing rates for natural movies, movies with modified spatio-temporal correlations as well as gratings. The activity patterns elicited by drifting gratings are qualitatively and quantitatively different from those elicited by natural stimuli and this difference arises from both spatial as well as temporal properties of the stimuli. Furthermore, both local field potentials and multiunit firing rates are most sensitive to the second-order statistics of the stimuli and not to their higher-order properties. Finally, responses to natural movies show a large variability over time because of activity fluctuations induced by rapid stimulus motion. We show that these fluctuations are not dependent on the detailed spatial properties of the stimuli but depend on their temporal jitter. These fluctuations are important characteristics of visual activity under natural conditions and impose limitations on the readout of possible differences in mean activity levels. PMID- 12750424 TI - Acoustic startle evokes bilaterally synchronous oscillatory EMG activity in the healthy human. AB - Despite animal evidence that the reticulospinal system is of major importance to movement, this motor pathway has remained relatively inaccessible to experimentation in the human. Consequently, little is known about its function in health and disease. Here, we use the acoustic startle response to demonstrate that one type of reticulospinal activity in the human is associated with a characteristic pattern of bilateral synchronization between motor units. Surface electromyography (EMG) was recorded from upper limb muscles in 15 healthy subjects during the reflex startle to unexpected acoustic stimulation, voluntary movements mimicking the startle and during sustained voluntary tonic contraction. Frequency analysis demonstrated autospectral peaks at approximately 14 Hz in deltoid and biceps muscles only during the startle reflex. Similarly, coherence spectra of the EMG recorded between homologous proximal upper limb muscles demonstrated a peak centered approximately 12-16 Hz during reflex startles. Coherence in the 10- to 20-Hz band was significantly greater in the startle reflex than during voluntary sham startles or voluntary tonic contraction for deltoid, but not first dorsal interosseous, muscles. The coherence at 10-20 Hz between EMGs from homologous muscles represents a potential surrogate measure of reticulospinal activity that may be useful in determining the contribution of the reticulospinal system to different types of movement in health and disease. PMID- 12750425 TI - Functional roles of an ERG current isolated in cerebellar Purkinje neurons. AB - Transcripts encoding ERG potassium channels are expressed by most neurons of the CNS. By patch-clamp whole cell recording from Purkinje neurons in slices of young (5-9 days old) mouse cerebellum we have been able to isolate a tail current [IK(ERG)] with the same characteristics as previously described for ERG channels. In zero external Ca2+ and high K+ (40 mM) the V1/2 of activation was -50.7 mV, the V1/2 of inactivation was -70.6 mV, and the deactivation rate was double exponential and voltage dependent. IK(ERG) was 93.0% blocked by WAY-123,398 (1 microM) and 78.2% by haloperidol (2 microM). The role of IK(ERG) on evoked firing was studied in adult mice, where WAY-123,398 application decreased the first spike latency, increased the firing frequency, and suppressed the frequency adaptation. However, the shape of individual action potentials was not affected. Stimulation of presynaptic climbing fibers evoked the Purkinje neuron "complex spike," composed of an initial spike and several spikelets. IK(ERG) block caused an increase of the number of spikelets of the "complex spike." These data show, for the first time, an IK(ERG) in a neuron of the CNS, the cerebellar Purkinje neuron, and indicate that such a current is involved in the control of membrane excitability, firing frequency adaptation, and in determining the effects of the climbing fiber synapse. PMID- 12750426 TI - Use-dependent shift from inhibitory to excitatory GABAA receptor action in SP-O interneurons in the rat hippocampal CA3 area. AB - Cortical inhibitory interneurons set the pace of synchronous neuronal oscillations implicated in synaptic plasticity and various cognitive functions. The hyperpolarizing nature of inhibitory postsynaptic potentials (IPSPs) in interneurons has been considered crucial for the generation of oscillations at beta (15-30 Hz) and gamma (30-100 Hz) frequency. Hippocampal basket cells and axo axonic cells in stratum pyramidale-oriens (S-PO) play a central role in the synchronization of the local interneuronal network as well as in pacing of glutamatergic principal cell firing. A lack of conventional forms of plasticity in excitatory synapses onto interneurons facilitates their function as stable neuronal oscillators. We have used gramicidin-perforated and whole cell clamp recordings to study properties of GABAAR-mediated transmission in CA3 SP-O interneurons and in CA3 pyramidal cells in rat hippocampal slices during electrical 5- to 100-Hz stimulation and during spontaneous activity. We show that GABAergic synapses onto SP-O interneurons can easily switch their mode from inhibitory to excitatory during heightened activity. This is based on a depolarizing shift in the GABAA reversal potential (EGABA-A), which is much faster and more pronounced in interneurons than in pyramidal cells. We also found that the shift in interneuronal function was frequency dependent, being most prominent at 20- to 40-Hz activation of the GABAergic synapses. After 40-Hz tetanic stimulation (100 pulses), GABAA responses remained depolarizing for approximately 45 s in the interneurons, promoting bursting in the GABAergic network. Hyperpolarizing EGABA-A was restored >60 s after the stimulus train. Similar but spontaneous GABAergic bursting was induced by application of 4 aminopyridine (100 microM) to slices. A shift to depolarizing IPSPs by the GABAAR permeant weak acid anion formate provoked interneuronal population bursting, supporting the role of GABAergic excitation in burst generation. Furthermore, depolarizing GABAergic potentials and synchronous interneuronal bursting were enhanced by pentobarbital (100 microM), a positive allosteric modulator of GABAARs, and were blocked by picrotoxin (100 microM). Intriguingly, GABAergic bursts displayed short (<1 s) oscillations at 15-40 Hz, even though only depolarizing GABAA responses were seen in the SP-O interneurons. This beta-gamma rhythmicity in the interneuron network was dependent on electrotonic coupling, and was abolished by blockade of gap junctions with carbenoxolone (200 microM). Results here implicate the rapid activity-dependent degradation of hyperpolarizing IPSPs in SP-O interneurons in setting the temporal limits for a given interneuron to participate in beta-gamma oscillations synchronized by GABAergic synapses. Furthermore, they imply that mutual GABAergic excitation provided by interneurons may be an integral part in the function of neuronal networks. We suggest that the use-dependent change in EGABA-A could represent a form of short-term plasticity in interneurons promoting coherent and sustained activation of local GABAergic networks. PMID- 12750427 TI - A differential pharmacokinetic analysis of the erythropoietin receptor population in newborn and adult sheep. AB - Strong evidence indicates that erythropoietin (Epo) is eliminated via Epo receptors (EpoR). Epo receptors may be classified as erythropoietic receptors that are largely located on erythroid progenitor cells in the bone marrow (BM) and nonerythropoietic receptors present in most tissues. Epo's elimination kinetics was studied using a very sensitive tracer interaction method (TIM) before and after chemical ablation of BM as an indirect way of evaluating the EpoR through an assortment of pharmacokinetic parameters (VM, KM, K, and CL) used in differentiating the EpoR population in newborn and adult sheep. TIM identified a parallel nonlinear Michaelis-Menten (VM and KM), and linear (K) elimination pathway and found the latter pathway to be significantly (p < 0.01) more dominant in lamb: K/(VM/KM + K) = 0.309 (25.3) versus 0.0895 (18.4) mean (CV%) lambs versus adult sheep. The significantly (p < 0.01) larger total clearance found for lambs indicates a larger nonhematopoietic tissue clearance of Epo (CL = 118 (10.9) ml/h/kg versus 67.8 (19.3) lamb versus adult sheep). The VM/KM ratio for the nonlinear pathway was not found to be significantly different (p > 0.05) between newborn and adults with values of 1.10 (15.8) and 1.30 (3.81) h-1, respectively. We proposed the hypothesis that the linear pathway is via nonhematopoietic EpoR. Assuming that Epo's elimination largely depends not only on erythropoietic EpoR but also on nonhematopoietic EpoR, this work shows a significant difference in the relative proportions of the two EpoR populations in lamb and adult sheep. The larger dominance of the nonhematopoietic EpoR in lamb supports the hypothesis that these receptors are more needed in early life, e.g., providing neuroprotection from perinatal hypoxemic-ischemic episodes. PMID- 12750429 TI - Role of cysteine S-conjugate beta-lyase in the metabolism of cisplatin. AB - Cisplatin is nephrotoxic, but the mechanism by which cisplatin kills renal proximal tubule cells is not well defined. Inhibition of gamma-glutamyl transpeptidase or pyridoxal 5'-phosphate (PLP)-dependent enzymes blocks the nephrotoxicity. Our hypothesis is that cisplatin is metabolized to a renal toxin through a platinum-glutathione conjugate to a reactive sulfur-containing compound. The final step in this bioactivation is the conversion of a platinum cysteine S-conjugate to a reactive thiol by a PLP-dependent cysteine S-conjugate beta-lyase. LLC-PK1 cells, a proximal tubule cell line with low cysteine S conjugate beta-lyase activity, are used to study cisplatin nephrotoxicity. We proposed that the beta-elimination reaction catalyzed by cysteine S-conjugate beta-lyase is the rate-limiting step in the metabolism of cisplatin to a toxin in these cells. In this study, LLC-PK1 cells were transfected with human glutamine transaminase K, which catalyzes the beta-elimination reaction. Cisplatin was significantly more toxic in confluent monolayers of cells with increased cysteine S-conjugate beta-lyase activity. In contrast, carboplatin, a non-nephrotoxic derivative of cisplatin, was 20-fold less toxic than cisplatin in confluent cells, and its toxicity was not altered by overexpression of cysteine S-conjugate beta-lyase. We propose that carboplatin is not nephrotoxic because it is not metabolized through this pathway. Dividing cells were more sensitive to both cisplatin and carboplatin toxicity. Overexpression of cysteine S-conjugate beta lyase activity had no effect on the toxicity of either drug. These data demonstrate that cisplatin kills quiescent renal cells by a mechanism that is distinct from the mechanism by which it kills dividing cells and that the renal toxicity of cisplatin is dependent on cysteine S-conjugate beta-lyase activity. PMID- 12750428 TI - Steroids modulate N-methyl-D-aspartate-stimulated [3H] dopamine release from rat striatum via sigma receptors. AB - Steroids have been proposed as endogenous ligands at sigma receptors. In the current study, we examined the ability of steroids to regulate N-methyl-d aspartate (NMDA)-stimulated [3H]dopamine release from slices of rat striatal tissue. We found that both progesterone and pregnenolone inhibit [3H]dopamine release in a concentration-dependent manner similarly to prototypical agonists, such as (+)-pentazocine. The inhibition seen by both progesterone and pregnenolone exhibits IC50 values consistent with reported Ki values for these steroids obtained in binding studies, and was fully reversed by both the sigma1 antagonist 1-(cyclopropylmethyl)-4-2'-4"flurophenyl)-2'oxoethyl)piperidine HBr (DuP734) and the sigma2 antagonist 1'-[4-[1-(4-fluorophenyl)-1-H-indol-3-yl]-1 butyl]spiro[iso-benzofuran-1(3H), 4'piperidine] (Lu28-179). Lastly, to determine whether a protein kinase C (PKC) signaling system might be involved in the inhibition of NMDA-stimulated [3H]dopamine release, we tested the PKCbeta selective inhibitor 5,21:12,17-dimetheno-18H-dibenzo[i,o]pyrrolo[3,4 - 1][1,8]diacyclohexadecine-18,20(19H)-dione,8-[(dimethylamino)methyl] 6,7,8,9,10,11-hexahydro-monomethanesulfonate (9Cl) (LY379196) against both progesterone and pregnenolone. We found that LY379196 at 30 nM reversed the inhibition of release by both progesterone and pregnenolone. These findings support steroids as candidates for endogenous ligands at sigma receptors. PMID- 12750430 TI - Rat hepatic CYP2E1 is induced by very low nicotine doses: an investigation of induction, time course, dose response, and mechanism. AB - CYP2E1 is an ethanol- and drug-metabolizing enzyme that can also activate procarcinogens and hepatotoxicants and generate reactive oxygen species; it has been implicated in the pathogenesis of liver diseases and cancer. Cigarette smoke increases CYP2E1 activity in rodents and in humans and we have shown that nicotine (0.1-1.0 mg/kg s.c. x 7 days) increases CYP2E1 protein and activity in the rat liver. In the current study, we have shown that the induction peaks at 4 h postnicotine (1 mg/kg s.c. x 7 days) treatment and recovers within 24 h. No induction was observed after a single injection, and 18 days of treatment did not increase the levels beyond that found at 7 days. We found that CYP2E1 is induced by very low doses of chronic (x 7 days) nicotine with an ED50 value of 0.01 mg/kg s.c.; 0.01 mg/kg in a rat model results in peak cotinine levels (nicotine metabolite) similar to those found in people exposed to environmental tobacco smoke (passive smokers; 2-7 ng/ml). Previously, we have shown no change in CYP2E1 mRNA, and our current mechanistic study indicates that nicotine does not regulate CYP2E1 expression by protein stabilization. We postulated that a nicotine metabolite could be causing the induction but found that cotinine (1 mg/kg x 7 days) did not increase CYP2E1. Our findings indicate that nicotine increases CYP2E1 at very low doses and may enhance CYP2E1-related toxicity in smokers, passive smokers, and people treated with nicotine (e.g., smokers, patients with Alzheimer's disease, ulcerative colitis or Parkinson's disease). PMID- 12750431 TI - Characterization of blood-brain barrier permeability to PYY3-36 in the mouse. AB - Peptide YY3-36 (PYY) has emerged as an important signal in the gut-brain axis, with peripherally administered PYY affecting feeding and brain function. For these effects to be direct, PYY would have to cross the blood-brain barrier (BBB). Here, we determined the permeability of the BBB to PYY radioactively labeled with 131I (I-PYY). Multiple-time regression analysis showed the unidirectional influx rate (Ki) from blood-to-brain for I-PYY to be 0.49 +/- 0.19 microl/g-min, a rate similar to that previously measured for leptin. Influx was not inhibited by 1 microg/mouse of unlabeled PYY, suggesting PYY crosses the BBB by transmembrane diffusion. About 0.176% of the i.v.-injected dose of I-PYY was taken up by brain, an amount similar to that for other peptides important in gut brain communication. Capillary depletion showed that 69% of I-PYY crossed the BBB to enter the parenchymal space of the brain, and high-performance liquid chromatography demonstrated that the radioactivity in this space represented intact I-PYY. After intracerebroventricular injection, I-PYY crossed from brain to blood by the mechanism of bulk flow. We conclude that PYY crosses in both the blood-to-brain and brain-to-blood directions by nonsaturable mechanisms. Passage across the BBB provides a mechanism by which blood-borne PYY can affect appetite and brain function. PMID- 12750432 TI - The novel melatonin agonist agomelatine (S20098) is an antagonist at 5 hydroxytryptamine2C receptors, blockade of which enhances the activity of frontocortical dopaminergic and adrenergic pathways. AB - Agomelatine (S20098) displayed pKi values of 6.4 and 6.2 at native (porcine) and cloned, human (h)5-hydroxytryptamine (5-HT)2C receptors, respectively. It also interacted with h5-HT2B receptors (6.6), whereas it showed low affinity at native (rat)/cloned, human 5-HT2A (<5.0/5.3) and 5-HT1A (<5.0/5.2) receptors, and negligible (<5.0) affinity for other 5-HT receptors. In antibody capture/scintillation proximity assays, agomelatine concentration dependently and competitively abolished h5-HT2C receptor-mediated activation of Gq/11 and Gi3 (pA2 values of 6.0 and 6.1). As measured by [3H]phosphatidylinositol depletion, agomelatine abolished activation of phospholipase C by h5-HT2C (pKB value of 6.1) and h5-HT2B (pKB value of 6.6) receptors. In vivo, it dose dependently blocked induction of penile erections by the 5-HT2C agonists (S)-2-(6-chloro-5 fluoroindol-1-yl)-1-methylethylamine (Ro60,0175) and 1-methyl-2-(5,8,8-trimethyl 8H-3-aza-cyclopenta[a]inden-3-yl) ethylamine (Ro60,0332). Furthermore, agomelatine dose dependently enhanced dialysis levels of dopamine in frontal cortex of freely moving rats, whereas they were unaffected in nucleus accumbens and striatum. Although the electrical activity of ventrotegmental dopaminergic neurons was unaffected agomelatine, it abolished their inhibition by Ro60,0175. Extracellular levels of noradrenaline in frontal cortex were also dose dependently enhanced by agomelatine in parallel with an acceleration in the firing rate of adrenergic cell bodies in the locus coeruleus. These increases in noradrenaline and dopamine levels were unaffected by the selective melatonin antagonist N-[2-(5-ethyl-benzo[b]thien-3-yl)ethyl] acetamide (S22153) and likely flect blockade of 5-HT2C receptors inhibitory to frontocortical dopaminergic and adrenergic pathways. Correspondingly, distinction to agomelatine, melatonin showed negligible activity 5-HT2C receptors and failed to modify the activity of adrenergic and dopaminergic pathways. In conclusion, in contrast to melatonin, agomelatine behaves as an antagonist at 5-HT2B and 5-HT2C receptors: blockade of the latter reinforces frontocortical adrenergic and dopaminergic transmission. PMID- 12750433 TI - Dissection of the anti-inflammatory effect of the core and C-terminal (KPV) alpha melanocyte-stimulating hormone peptides. AB - In this study, we analyzed the anti-inflammatory effects of alpha-melanocyte stimulating hormone (MSH)11-13 (KPV) in comparison with other MSH peptides in a model of crystal-induced peritonitis. Systemic treatment of mice with KPV, alpha MSH, the core melanocortin peptide His-Phe-Arg-Trp, and the melanocontin receptor 3/4 agonist Ac-Nle4-c[Asp5,d-Phe7,Lys10]NH2 ACTH4-10 (MTII) but not the selective MC1-R agonist H-Ser-Ser-Ile-Ile-Ser-His-Phe-Arg-Trp-Gly-Lys-Pro-Val-NH2 (MS05) resulted in a significant reduction in accumulation of polymorphonuclear leukocyte in the peritoneal cavity. The antimigratory effect of KPV was not blocked by the MC3/4-R antagonist Ac-Nle4-c[Asp5,d-2Nal7,Lys10]NH2 ACTH4-10 (SHU9119). In vitro, macrophage activation, determined as release of KC and interleukin (IL)-1beta was inhibited by alpha-MSH and MTII but not by KPV. Furthermore, macrophage activation by MTII led to an increase in cAMP accumulation, which was attenuated by SHU9119, whereas KPV failed to increase cAMP. The anti-inflammatory properties of KPV were also evident in IL-1beta induced peritonitis inflammation and in mice with a nonfunctional MC1-R (recessive yellow e/e mice). In conclusion, these data highlight that the C terminal MSH peptide KPV exhibits an anti-inflammatory effect that is clearly different from that of the core MSH peptides. KPV is unlikely to mediate its effects through melanocortin receptors but is more likely to act through inhibition of IL-1beta functions. PMID- 12750434 TI - Mu-opioid-induced desensitization of opioid receptor-like 1 and mu-opioid receptors: differential intracellular signaling determines receptor sensitivity. AB - Mu-Opioid receptors have been shown to contribute to orphanin FQ/nociceptin (OFQ/N)-mediated analgesia and hyperalgesia, indicating that both pro- and antinociceptive actions of OFQ/N are influenced by mu-opioid receptors. A 60-min activation of mu-or opioid receptor-like 1 (ORL1) opioid receptors natively expressed in BE(2)-C human neuroblastoma cells desensitized both mu- and ORL1 receptor-mediated inhibition of cAMP accumulation. The mechanism(s) of OFQ/N mediated mu and ORL1 cross talk involves the conventional protein kinase C isozyme, PKC-alpha, and G protein-coupled receptor kinases (GRKs) 2 and 3. Unlike OFQ/N-mediated desensitization of ORL1 and mu-opioid receptors, [d-Ala2,N-Me Phe4,Gly5-ol]-enkephalin (DAMGO)-mediated ORL1 desensitization in BE(2)-C cells is PKC-independent. However, DAMGO (1 microM) pretreatment increased membrane levels of GRK2 and GRK3, indicating their translocation to the membrane upon activation. This suggests that DAMGO activation of mu-opioid receptors results in GRK2 and GRK3 inactivation of ORL1 upon challenge with OFQ/N. Antisense, but not sense, DNA selectively targeting GRK2 or GRK3 blocks DAMGO-mediated mu- and ORL1 desensitization, respectively. However, in SH-SY5Y neuroblastoma cells, DAMGO failed to desensitize ORL1 or alter membrane PKC-alpha or GRK levels. Instead, DAMGO stimulated PKC-epsilon translocation to the cell membrane and produced micro-receptor desensitization. These results indicate that acute exposure to mu receptor agonists can regulate ORL1 function, but the ability to do so varies from cell type to cell type. These results also confirm the existence of multiple signaling mechanisms for mu-opioid receptors and the importance of these mechanisms for mu-receptor-mediated-heterologous effects. PMID- 12750435 TI - Neocarzinostatin induces an effective p53-dependent response in human papillomavirus-positive cervical cancer cells. AB - Human papillomavirus (HPV) E6 viral oncoprotein plays an important role during cervical carcinogenesis. This oncoprotein binds the tumor suppressor protein p53, leading to its degradation via the ubiquitin-proteasome pathway. Therefore, it is generally assumed that in HPV-positive cancer cells p53 function is completely abolished. Nevertheless, recent findings suggest that p53 activity can be recovered in cells expressing endogenous E6 protein. To investigate whether p53 dependent functions controlling genome integrity, cell proliferation, and apoptosis can be reactivated in cervical cancer cells, we examined the capacity of HeLa, INBL, CaSki, C33A, and ViBo cell lines to respond to neocarzinostatin (NCS), a natural product which induces single- and double-strand breaks in DNA. We found that NCS treatment inhibits cellular proliferation through G2 cell cycle arrest and apoptosis induction. This effect was preceded by nuclear accumulation of p53 protein and by an increase of p21 transcripts. Although apoptosis was blocked in ViBo cells (HPV-negative), nuclear accumulation of transcriptionally active p53 and inhibition of cell proliferation are observed after NCS treatment. These results suggest that HPV-positive cervical cancer cells are capable of responding efficiently to DNA damage provoked by NCS treatment through a p53 dependent pathway in spite of the presence of E6 protein. PMID- 12750436 TI - L-selegiline potentiates the cellular poly(ADP-ribosyl)ation response to ionizing radiation. AB - DNA strand breaks induced by alkylating agents, oxidants, or ionizing radiation trigger the covalent modification of nuclear proteins with poly(ADP-ribose), which is catalyzed for the most part by poly(ADP-ribose) polymerase-1 and plays a role in DNA base-excision repair. Poly(ADP-ribosyl)ation capacity of mononuclear blood cells correlates positively with life span of mammalian species. Here, we show that l-selegiline, an anti-Parkinson drug with neuroprotective activity and life span-extending effect in laboratory animals, can potentiate gamma-radiation induced poly(ADP-ribose) formation in intact cells. COR4 hamster cells were incubated with l-selegiline (50 nM) for various time periods, followed by gamma irradiation (45 Gy). Quantification of cellular poly(ADP-ribose) levels at 10 min after starting the irradiation revealed significant increases (up to 1.8-fold) in cells preincubated with the drug for 8 h to 7 days compared with drug-free irradiated controls. There was no selegiline-induced change in poly(ADP-ribose) levels of unirradiated cells nor in basal or radiation-induced DNA strand breaks, respectively. Surprisingly, poly(ADP-ribose) polymerase-1 protein was down regulated by l-selegiline treatment. Addition of l-selegiline to purified poly(ADP-ribose) polymerase-1 did not alter enzymatic activity. In conclusion, the results of the present study identify a novel intervention to potentiate the cellular poly(ADP-ribosyl)ation response. We hypothesize that the effect of l selegiline is due to modulation of accessory proteins regulating poly(ADP-ribose) polymerase-1 activity and that increased cellular poly- (ADP-ribosyl)ation capacity may contribute to the neuroprotective potential and/or life span extension afforded by l-selegiline. PMID- 12750437 TI - Gene expression in the human intestine and correlation with oral valacyclovir pharmacokinetic parameters. AB - The transport of valacyclovir, the l-valyl ester of acyclovir, has been suggested to be mediated by several carrier-mediated pathways in cell culture and animal models. The role and importance of these transporters in modulating valacyclovir absorption in humans has not been determined, however. Recent advances in genomic technology have facilitated the rapid and simultaneous determination of global mRNA expression profiles for thousands of genes in tissue biopsies directly associated with the absorption process, thereby dramatically increasing the value of studies in humans. In this article, we describe correlations of pharmacokinetic parameters following oral valacyclovir or acyclovir administration with expression levels of intestinal genes in humans. Highly positive and significant correlations were observed with 4F2hc, an activator of cation-preferring amino acid transport systems, and human oligopeptide transporter (HPT1), an oligopeptide transporter expressed at higher levels in the human intestine compared with oligopeptide transporter (PEPT1). The validation of HPT1 microarray data with reverse transcription-polymerase chain reaction and the enhanced valacyclovir uptake in HeLa/HPT1 cells suggest that the role of HPT1 in transport of peptides and peptidomimetics drugs needs to be examined in more detail. The interrelation of 4F2hc and HPT1 in transport may be of interest. No significant correlations of valacyclovir pharmacokinetic parameters with PEPT1 and with organic cation or anion transporter expression levels were observed. The highly negative correlations observed with known efflux pumps such as MDR1 (P glycoprotein) and MRP2 (cMOAT), as well as with the CYP450 IIIA subfamily may indicate that these proteins may regulate the cellular accumulation and metabolism of acyclovir. PMID- 12750438 TI - A growth hormone secretagogue prevents ischemic-induced mortality independently of the growth hormone pathway in dogs with chronic dilated cardiomyopathy. AB - To determine the functional role of growth hormone (GH) secretagogue in myocardium with ischemia, left ventricular (LV) pressure gauge, wall thickness crystals, coronary occluder, pacers, and catheters were implanted in 26 dogs. Beginning 1 week after ventricular pacing (240 beats/min) was initiated, dogs were treated (s.c.) with GH releasing peptide-6 (GHRP-6, n = 8, 0.2 mg/kg/day), GH (n = 7, 0.06 mg/kg/day), or vehicle (n = 11). Two weeks of pacing was associated with similar decreases in LV pressure, rate of change of LV pressure, systolic wall thickening (WT), and an increase in left atrial pressure in all groups. Coronary artery occlusion (CAO) resulted in a similar loss of WT in ischemic regions, which did not recover during reperfusion period in all groups. WT in nonischemic regions, however, was enhanced in the GHRP-6 group compared with the GH and vehicle groups, e.g., increase of WT after 1 h of reperfusion was greater (p <0.05) in the GHRP-6 (+53 +/- 8%) than in the GH (+14 +/- 12%) or (+14 +/- 6%). There were no differences in myocardial blood flow, hemodynamics, or arrhythmic beats among all groups during CAO and reperfusion periods. Strikingly, no dogs in the GHRP-6 group died during CAO, whereas the survival rates for GH and vehicle groups were 57 and 55%, respectively. Our data demonstrate, for the first time, that chronic therapy with a GH secretagogue prevents sudden death in dogs with dilated cardiomyopathy subjected to acute ischemia. This seems to be related to an enhanced nonischemic compensatory mechanism mediated by the GH secretagogue receptors rather than via the GH/insulin growth factor-1 pathway. PMID- 12750439 TI - A lymphocyte-generated fragment of vasoactive intestinal peptide with VPAC1 agonist activity and VPAC2 antagonist effects. AB - Vasoactive intestinal peptide receptors 1 (VPAC1) and 2 (VPAC2) have been identified in humans. Cell lines expressing only VPAC1 (HT-29) or VPAC2 (Molt-4b) were identified using real-time reverse transcriptase polymerase chain reaction. Vasoactive intestinal peptide (VIP) and related peptides, VIP-6-28, VIP4-28, and VIP10-28, previously isolated from cultures of human leukocytes, were evaluated for their ability to bind to VPAC1 and VPAC2 and to increase the levels of cAMP in HT-29 and Molt-4b cells. VIP bound to membranes of HT-29 colon carcinoma cells and Molt-4b lymphoblasts with high affinity (KD = 1.6 +/- 0.2 and 1.7 +/- 0.9 nM, respectively). VIP4-28 also demonstrated high-affinity binding (KD = 1.7 +/- 0.2 and 1.7 +/- 0.7 nM in HT-29 and Molt-4b, respectively). VIP and VIP4-28 are potent VPAC1 agonists, inducing maximal 200- and 400-fold increases in cAMP, respectively. VIP demonstrated weak VPAC2 agonist activity, inducing a maximal 14 fold increase in cAMP. VIP4-28 had no VPAC2 agonist activity but demonstrated potent VPAC2 antagonist activity. VIP4-28 inhibited VPAC2-mediated increases in cAMP in Molt-4b cells up to 95%, but had no antagonistic effect on VPAC1. Lymphoblasts did not hydrolyze VIP4-28 to a form with VPAC1 antagonist activity. VIP4-28 thus is a lymphocyte-generated VIP fragment with potent agonist activity for VPAC1 and potent antagonist activity for VPAC2. PMID- 12750440 TI - Antinociceptive activity of the N-methyl-D-aspartate receptor antagonist N-(2 Indanyl)-glycinamide hydrochloride (CHF3381) in experimental models of inflammatory and neuropathic pain. AB - N-(2-Indanyl)-glycinamide hydrochloride (CHF3381) is a novel low-affinity, noncompetitive N-methyl-d-aspartate receptor antagonist. The current study compared the antinociceptive effects of CHF3381 with those of gabapentin and memantine in in vitro and in vivo models of pain. In isolated rat spinal cord, CHF3381 and memantine, but not gabapentin, produced similar inhibition of the wind-up phenomenon. CHF3381 suppressed the maintenance of carrageenan-induced thermal and mechanical hyperalgesia in the rat with a minimum significantly effective dose (MED) of 30 mg/kg p.o. Memantine produced a partial reversal of both thermal and mechanical hyperalgesia (MED = 10 and 15 mg/kg i.p., respectively). Gabapentin reversed mechanical hyperalgesia (MED = 10 mg/kg s.c.), but did not affect thermal hyperalgesia. In the mouse formalin test, CHF3381 and memantine preferentially inhibited the late phase (MED = 30 and 20 mg/kg i.p., respectively); gabapentin inhibited only the late phase (MED = 30 mg/kg s.c.). Unlike morphine, CHF3381 chronic administration was not accompanied by the development of tolerance in the formalin test. Furthermore, morphine tolerance did not cross-generalize to CHF3381. In rats with a sciatic nerve injury, CHF3381 relieved both cold and mechanical allodynia (MED = 100 mg/kg p.o.). In contrast, memantine was inactive. Gabapentin blocked cold allodynia (MED = 30 mg/kg s.c.), but had marginal effects on mechanical allodynia. In diabetic neuropathy, CHF3381 reversed mechanical hyperalgesia (MED = 50 mg/kg p.o.). Memantine (15 mg/kg i.p.) produced an antinociceptive effect, whereas gabapentin (100 mg/kg p.o.) had no significant effect. Thus, CHF3381 may be useful for the therapy of peripheral painful neuropathies. PMID- 12750442 TI - Mitochondrial regulation of apoptosis. AB - Mitochondria play a central part in cellular survival and apoptotic death. These processes are highly regulated by pro- and antiapoptotic Bcl-2 superfamily members. A key feature within apoptosis cascades is disruption of mitochondrial transmembrane potential and apoptogenic protein release, caused by opening of the permeability transition pore (PT). New data, however, indicate that mitochondrial apoptosis may occur without PT involvement. PMID- 12750441 TI - Pilot intervention: aerosolized adrenomedullin reduces pulmonary hypertension. AB - In pulmonary hypertension, systemic infusion of adrenomedullin (ADM), a potent vasodilator peptide, leads to pulmonary vasodilatation. However, systemic blood pressure declines alike. The present study investigated the effect of aerosolized ADM on pulmonary arterial pressure in surfactant-depleted newborn piglets with pulmonary hypertension. Animals randomly received aerosolized ADM (ADM, n = 6), aerosolized ADM combined with intravenous application of NG-nitro-l-arginine methylester to inhibit nitric-oxide (NO) synthases (ADM + l-NAME, n = 5), or aerosolized normal saline solution (control, n = 6). Aerosol therapy was performed in 30-min intervals for 5 h. After a total experimental period of 8 h, mRNA expression of endothelial and inducible NO synthase and endothelin-1 (ET-1) in lung tissue was quantified using TaqMan real-time polymerase chain reaction. Aerosolized ADM reduced mean pulmonary artery pressure (MPAP) compared with control (p < 0.001; at the end of the study, Delta-MPAP -13.5 +/- 1.4 versus -6.2 +/- 2.4 mm Hg). PaO2 significantly increased in the ADM (DeltaPaO2 243.3 mm Hg) and the ADM + l-NAME group (DeltaPaO2 217.4 mm Hg) compared with the control group (DeltaPaO2 82.9 mm Hg; p < 0.001). Aerosolized ADM did not influence mean systemic arterial pressure (baseline 63.2 +/- 2.7 versus end of the study 66.3 +/ 6.5 mm Hg; not significant). NO synthases gene expressions were 20 to 30% lower with ADM compared with control. ET-1 gene expression was significantly reduced (>50%) after ADM aerosol therapy (p < 0.001). Aerosolized adrenomedullin significantly reduced MPAP without lowering the systemic arterial pressure and improved profoundly the arterial oxygen tension. This effect seems to be mediated at least in part by the reduction of ET-1. PMID- 12750443 TI - Nonredundant gap junction functions. AB - The need for molecular heterogeneity of gap junction channel proteins in vivo has been enigmatic. Recently, functional replacement of one channel gene with another in mice and flies has revealed that cellular health depends not simply on gap junction communication but also requires the correct type of intercellular channel subunit. PMID- 12750445 TI - Immune adherence revisited: novel players in an old game. AB - Erythrocytes bind immune complexes (ICs) composed of antibodies binding their respective antigen (e.g., bacteria, parasites, viruses, or autoantigen) plus complement proteins via complement receptors [immune adherence (IA)]. In vivo studies have shown that erythrocytes act as an inert shuttle, targeting ICs to fixed macrophages in liver and spleen. Here we outline established and emerging implications of IA in health and disease. PMID- 12750444 TI - New roles for connexons. AB - Connexons or gap junction hemichannels are large, nonselective ion channels that reside in the nonjunctional plasma membrane before their assembly into gap junction channels. Increasing evidence suggests that these channels can open under certain conditions and may participate in a number of cellular processes, including the release of small metabolites such as ATP and NAD(+), which are involved in paracrine signaling. PMID- 12750446 TI - Central nervous system processing of human visceral pain in health and disease. AB - To understand the pathophysiology of anomalous pain in functional gastrointestinal disorders, we must increase our understanding of how the central nervous system processes visceral pain. Over the past decade, novel application of functional brain imaging and electrophysiological techniques has given us the opportunity to study these processes in humans, and this review summarizes the current body of knowledge. PMID- 12750447 TI - Vesicle-mediated restoration of a plasmalemmal barrier in severed axons. AB - Ca(2+)-induced endocytotic vesicles undergo protein-mediated interactions to restore a selectively permeable barrier and propagated action potentials in severed invertebrate giant axons. Similar barrier-restoration phenomena observed in cultured mammalian cells with transected neurites suggest that cellular/molecular mechanisms that repair plasmalemmal damage are phylogenetically conserved. PMID- 12750449 TI - Functional modulation of the sodium pump: the regulatory proteins "Fixit". AB - Proteins of the FXYD family act as tissue-specific regulators of the Na-K-ATPase. They are small hydrophobic type I proteins with a single-transmembrane span containing an extracellular invariant FXYD sequence. FXYD proteins are not an integral part of the Na-K-ATPase but function to modulate its catalytic properties by molecular interactions with specific Na-K-ATPase domains. PMID- 12750450 TI - Pathophysiological basis of smoke inhalation injury. AB - Smoke inhalation injury results in serious respiratory failure. When smoke inhalation injury is combined with burn injury or pneumonia, the physiological responses are different and more severe than those of smoke inhalation injury alone. Treatment strategies should be planned based on these pathophysiological aspects. PMID- 12750451 TI - Chemotropic molecules: guides for axonal pathfinding and cell migration during CNS development. AB - Different molecules (netrins, semaphorins, slits) with chemotropic functions and their receptors (neogenin, DCC, neuropilins, plexins, robos) have been identified that guide axons during development of the nervous system to establish the complex pattern of connections among a large number of neurons. These molecules have been recently identified to play a role in cell migration of the central nervous system during development. PMID- 12750452 TI - Disruption of glucocorticoid receptor exon 2 yields a ligand-responsive C terminal fragment that regulates gene expression. AB - Mice in which exon 2 of the glucocorticoid receptor (GR) has been disrupted [GR exon 2 knockout (GR2KO)] have been used as a model to study the requirement for this receptor in a number of biological systems. A recent report showed that these mice actually express a truncated ligand-binding GR fragment, prompting us to ask whether this mutation truly results in a glucocorticoid-insensitive phenotype. Based on cDNA microarray analysis of fetal thymocytes, we found that glucocorticoids were able to enhance or repress activation-induced gene expression in GR2KO and wild-type thymocytes to a similar degree. Moreover, although changes in gene expression induced by glucocorticoids alone were blunted, the expression of a substantial number of genes in GR2KO thymocytes was modulated by stimulation with glucocorticoids. Among these genes, as confirmed by quantitative real-time PCR, was the classic glucocorticoid-responsive gene glutamine synthetase as well as genes implicated in T cell development and function such as IL-7 receptor alpha-chain and glucocorticoid-induced leucine zipper (GIL2). Thus, the truncated C-terminal GR2KO product, which lacks the major transactivation domain, retains, to a large extent, the ability to regulate gene expression both positively and negatively in a ligand-responsive manner when expressed in vivo. PMID- 12750454 TI - Compensatory role of thyroid hormone receptor (TR) alpha 1 in resistance to thyroid hormone: study in mice with a targeted mutation in the TR beta gene and deficient in TR alpha 1. AB - Resistance to thyroid hormone (RTH) is caused by mutations of the thyroid hormone receptor beta (TR beta) gene. Almost all RTH patients are heterozygous with an autosomal dominant pattern of inheritance. That most are clinically euthyroid suggests a compensatory role of the TR alpha1 isoform in maintaining the normal functions of thyroid hormone (T3) in these patients. To understand the role of TR alpha1 in the manifestation of RTH, we compared the phenotypes of mice with a targeted dominantly negative mutant TR beta (TR betaPV) with or without TR alpha1. TR betaPV mice faithfully recapitulate RTH in humans in that these mice demonstrate abnormalities in the pituitary-thyroid axis and impairment in growth. Here we show that the dysregulation of the pituitary-thyroid axis was worsened by the lack of TR alpha1 in TR betaPV mice, and severe impairment of postnatal growth was manifested in TR betaPV mice deficient in TR alpha1. Furthermore, abnormal expression patterns of T3-target genes in TR betaPV mice were altered by the lack of TR alpha1. These results demonstrate that the lack of TR alpha1 exacerbates the manifestation of RTH in TR betaPV mice. Therefore, TR alpha1 could play a compensatory role in mediating the functions of T3 in heterozygous patients with RTH. This compensatory role may be especially crucial for postnatal growth. PMID- 12750453 TI - The role of hepatocyte nuclear factor-3 alpha (Forkhead Box A1) and androgen receptor in transcriptional regulation of prostatic genes. AB - Androgens and mesenchymal factors are essential extracellular signals for the development as well as the functional activity of the prostate epithelium. Little is known of the intraepithelial determinants that are involved in prostatic differentiation. Here we found that hepatocyte nuclear factor-3 alpha (HNF-3 alpha), an endoderm developmental factor, is essential for androgen receptor (AR) mediated prostatic gene activation. Two HNF-3 cis-regulatory elements were identified in the rat probasin (PB) gene promoter, each immediately adjacent to an androgen response element. Remarkably, similar organization of HNF-3 and AR binding sites was observed in the prostate-specific antigen (PSA) gene core enhancer, suggesting a common functional mechanism. Mutations that disrupt these HNF-3 motifs significantly abolished the maximal androgen induction of PB and PSA activities. Overexpressing a mutant HNF-3 alpha deleted in the C-terminal region inhibited the androgen-induced promoter activity in LNCaP cells where endogenous HNF-3 alpha is expressed. Chromatin immunoprecipitation revealed in vivo that the occupancy of HNF-3 alpha on PSA enhancer can occur in an androgen-depleted condition, and before the recruitment of ligand-bound AR. A physical interaction of HNF-3 alpha and AR was detected through immunoprecipitation and confirmed by glutathione-S-transferase pull-down. This interaction is directly mediated through the DNA-binding domain/hinge region of AR and the forkhead domain of HNF 3 alpha. In addition, strong HNF-3 alpha expression, but not HNF-3 beta or HNF-3 gamma, is detected in both human and mouse prostatic epithelial cells where markers (PSA and PB) of differentiation are expressed. Taken together, these data support a model in which regulatory cues from the cell lineage and the extracellular environment coordinately establish the prostatic differentiated response. PMID- 12750455 TI - Subtle lung nodules: influence of local anatomic variations on detection. AB - PURPOSE: To assess the influence of local anatomic noise on the detection of subtle lung nodules depicted on chest radiographs. MATERIALS AND METHODS: Six 8 x 8-cm lung regions were extracted from digital chest radiographs obtained in healthy subjects. Simulated nodules emulating the radiographic characteristics of subtle tissue-equivalent lesions 3.2-6.4 mm in diameter (equivalent to 0.1-0.4 mm in contrast-diameter product [CD]) were added to the images. On multiple renditions of each image, nodules were inserted at slightly different locations within 6 mm of the center; this process allowed different local background patterns to overlie the nodules. An observer detection study involving 15 experienced radiologists was performed. The authors performed analysis of variance and pairwise t test analyses to determine variations in nodule detectability related to nodule location and size on each image. RESULTS: Results indicated a strong correlation between nodule size and observer detection score and significant variation in nodule detectability as a function of location. Changes in nodule position caused observer score variations that were equivalent to the variation caused by an up to 185% change in nodule CD (78% average over all six images), an up to 68% change in diameter (32% average), and an up to 28% change in area under the receiver operating characteristic curve (Az) (14% average). CONCLUSION: Local anatomic variations surrounding and overlying a subtle lung nodule on a chest radiograph that are created by the projection of anatomic features in the thorax, such as ribs and pulmonary vessels, can greatly influence the detection of nodules, altering the Az by as much as 28%. PMID- 12750456 TI - Influence of irradiated volume on ureteral injury after intraoperative radiation therapy: experimental study in dogs. AB - PURPOSE: To investigate the role of irradiated volumes at intraoperative radiation therapy in the development of ureteral injury in dogs. MATERIALS AND METHODS: Sixteen beagle dogs were randomized to receive 30 Gy of intraoperative radiation therapy in the right ureter. Lead shielding ensured that different volumes were irradiated. Six dogs received a 4 x 12-cm field, five dogs a 4 x 8 cm field, and five dogs a 4 x 4-cm field. Follow-up included magnetic resonance (MR) imaging, clinical examination, and resting sequential renography. Twelve months after irradiation, the animals were killed, and autopsy was performed. Functional outcome was defined as MR imaging and renography findings and was evaluated statistically by using the Cochran-Armitage test at a.05 significance level. RESULTS: Twelve months after therapy, ureteral obstruction with consecutive hydronephrosis of the right kidney was observed in four of six animals that received the largest volume of irradiation. Two dogs that received the medium volume developed ureteral obstruction. None of the five dogs that received the smallest volume showed abnormal findings (P <.05). The irradiated parts of the ureters in all dogs showed abnormal histopathologic findings, such as fibrosis. CONCLUSION: The probability of ureteral obstruction following intraoperative radiation therapy increases with the irradiated partial volume of the ureter. PMID- 12750457 TI - Optimization of oral contrast agents for MR imaging of the small bowel. AB - Effect on small-bowel distention of additives to water as contrast agents for magnetic resonance (MR) imaging was assessed. Oral contrast agents included water and water in combination with mannitol, a bulk fiber laxative, locust bean gum, and a combination of mannitol and locust bean gum. Filling of the small bowel was quantified on coronal images obtained with two-dimensional true fast imaging with steady-state precession sequence; bowel diameters were measured. Ingestion of water with locust bean gum and mannitol provided the best distention of the small bowel. MR imaging of the small bowel with oral administration of water can be improved with addition of osmotic and nonosmotic substances that lead to decreased water resorption. PMID- 12750458 TI - Structure and function of small airways in smokers: relationship between air trapping at CT and airway inflammation. AB - PURPOSE: To use quantitative computed tomography (CT) to compare lung attenuation with both inflammatory infiltration and in vitro reactivity of peripheral airways in smokers scheduled to undergo lung resection for localized pulmonary lesions. MATERIALS AND METHODS: Attenuation was measured in nine ex-smokers, 13 current smokers, and eight nonsmoking control subjects by using CT with respiratory gating and a contour-tracing algorithm. After lung resection in smokers, peripheral bronchi were dissected and studied in terms of both inflammation (by using immunohistochemistry to examine glycolmethacrylate-embedded specimens) and mechanical activity (by using an isolated organ bath system). Comparisons between groups were made by using analysis of variance and subsequent unpaired t tests. Correlations were evaluated by using the Pearson coefficient and stepwise multiple regression analysis. RESULTS: The difference between inspiratory and expiratory attenuation was significantly higher in control subjects (-128 HU +/- 11 [SD]) than in ex-smokers (-77 HU +/- 10; P =.004) or current smokers (-67 HU +/- 11; P =.001). Cells infiltrating the smooth muscle increased with the decrease in expiratory attenuation (r = -0.46; P =.03) and the increase in inspiratory versus expiratory attenuation (r = 0.66; P =.001). Mast cell and neutrophil infiltration of smooth muscle was the most important factor in this relationship. Cellular infiltration of the smooth muscle increased with the decrease of in vitro relaxation response to salbutamol. CONCLUSION: In smokers, air trapping is correlated with inflammatory infiltration of the smooth muscle layer of small airways. PMID- 12750459 TI - Alveolar echinococcosis: MR findings in the liver. AB - PURPOSE: To clarify the magnetic resonance (MR) imaging findings of alveolar echinococcosis in the liver. MATERIALS AND METHODS: Thirty-five patients with 50 lesions histologically proven to be alveolar echinococcosis were evaluated with MR imaging. Lesions were assessed with regard to the distribution pattern of solid and cystic components and pattern of contrast material enhancement. RESULTS: Cystic components exhibited two patterns at T2-weighted MR imaging: small round cysts and large and/or irregular cysts. Forty-eight lesions (96%) contained small round cysts. Twenty-six lesions (52%) had large and/or irregular cysts. Forty-five lesions (90%) were associated with a solid component. MR imaging characteristics were categorized into five types: multiple small round cysts without a solid component (two lesions [4%], type 1), multiple small round cysts with a solid component (20 lesions [40%], type 2), a solid component surrounding large and/or irregular cysts with multiple small rounds cysts (23 lesions [46%], type 3), a solid component without cysts (two lesions [4%], type 4), and a large cyst without a solid component (three lesions [6%], type 5). In most cases (97%), contrast enhancement was weak. CONCLUSION: The MR findings of alveolar echinococcosis in the liver are multiple small round cysts with a weakly enhanced solid component. The cystic component can be a large and/or irregular lesion, and such lesions are depicted clearly at T2-weighted MR imaging. PMID- 12750460 TI - Can noise reduction filters improve low-radiation-dose chest CT images? Pilot study. AB - Effect of noise reduction filters on chest computed tomographic (CT) images acquired with 50% radiation dose reduction was evaluated. Two sets of images were acquired with multi-detector row CT at standard (220-280 mA) and 50% reduced (110 140 mA) tube current at the level of the carina. After postprocessing with six noise reduction filters, images were compared with baseline standard-dose images for noise, sharpness, and contrast in lungs, mediastinum, and chest wall. Quantitative image noise was measured in descending thoracic aorta. Modulation transfer functions were calculated from CT images of 50-microm wire. Noise reduction filters reduced image noise on low-radiation-dose chest CT images, with some compromise in image sharpness and contrast assessed qualitatively, and slightly altered modulation transfer function at higher spatial frequencies. PMID- 12750461 TI - Spherical aggregates of beta-amyloid (amylospheroid) show high neurotoxicity and activate tau protein kinase I/glycogen synthase kinase-3beta. AB - beta-Amyloid (Abeta) acquires toxicity by self-aggregation. To identify and characterize the toxic form(s) of Abeta aggregates, we examined in vitro aggregation conditions by using large quantities of homogenous, chemically synthesized Abeta1-40 peptide. We found that slow rotation of Abeta1-40 solution reproducibly gave self-aggregated Abeta1-40 containing a stable and highly toxic moiety. Examination of the aggregates purified by glycerol-gradient centrifugation by atomic force microscopy and transmission electron microscopy revealed that the toxic moiety is a perfect sphere, which we call amylospheroid (ASPD). Other Abeta1-40 aggregates, including fibrils, were nontoxic. Correlation studies between toxicity and sphere size indicate that 10- to 15-nm ASPD was highly toxic, whereas ASPD <10 nm was nontoxic. A positive correlation between the toxicity and ASPD >10 nm also appeared to exist when Abeta1-42 formed ASPD by slow rotation. However, Abeta1-42-ASPD formed more rapidly, killed neurons at lower concentrations, and showed approximately 100-fold-higher toxicity than Abeta1-40-ASPD. The toxic ASPD was associated with SDS-resistant oligomeric bands in immunoblotting, which were absent in nontoxic ASPD. Because the formation of ASPD was not disturbed by pentapeptides that break beta-sheet interactions, Abeta may form ASPD through a pathway that is at least partly distinct from that of fibril formation. Inhibition experiments with lithium suggest the involvement of tau protein kinase I/glycogen synthase kinase-3beta in the early stages of ASPD induced neurodegeneration. Here we describe the identification and characterization of ASPD and discuss its possible role in the neurodegeneration in Alzheimer's disease. PMID- 12750462 TI - D-serine and serine racemase are present in the vertebrate retina and contribute to the physiological activation of NMDA receptors. AB - d-serine has been proposed as an endogenous modulator of N-methyl-d-aspartate (NMDA) receptors in many brain regions, but its presence and function in the vertebrate retina have not been characterized. We have detected d-serine and its synthesizing enzyme, serine racemase, in the retinas of several vertebrate species, including salamanders, rats, and mice and have localized both constituents to Muller cells and astrocytes, the two major glial cell types in the retina. Physiological studies in rats and salamanders demonstrated that, in retinal ganglion cells, d-serine can enhance excitatory currents elicited by the application of NMDA, as well as the NMDA receptor component of light-evoked synaptic responses. Application of d-amino acid oxidase, which degrades d-serine, reduced the magnitude of NMDA receptor-mediated currents, raising the possibility that endogenous d-serine serves as a ligand for setting the sensitivity of NMDA receptors under physiological conditions. These observations raise exciting new questions about the role of glial cells in regulating the excitability of neurons through release of d-serine. PMID- 12750464 TI - Spontaneous mitotic homologous recombination at an enhanced yellow fluorescent protein (EYFP) cDNA direct repeat in transgenic mice. AB - A transgenic mouse has been created that provides a powerful tool for revealing genetic and environmental factors that modulate mitotic homologous recombination. The fluorescent yellow direct-repeat (FYDR) mice described here carry two different copies of expression cassettes for truncated coding sequences of the enhanced yellow fluorescent protein (EYFP), arranged in tandem. Homologous recombination between these repeated elements can restore full-length EYFP coding sequence to yield a fluorescent phenotype, and the resulting fluorescent recombinant cells are rapidly quantifiable by flow cytometry. Analysis of genomic DNA from recombined FYDR cells shows that this mouse model detects gene conversions, and based on the arrangement of the integrated recombination substrate, unequal sister-chromatid exchanges and repair of collapsed replication forks are also expected to reconstitute EYFP coding sequence. The rate of spontaneous recombination in primary fibroblasts derived from adult ear tissue is 1.3 +/- 0.1 per 106 cell divisions. Interestingly, the rate is approximately 10 fold greater in fibroblasts derived from embryonic tissue. We observe an approximately 15-fold increase in the frequency of recombinant cells in cultures of ear fibroblasts when exposed to mitomycin C, which is consistent with the ability of interstrand crosslinks to induce homologous recombination. In addition to studies of recombination in cultured primary cells, the frequency of recombinant cells present in skin was also measured by direct analysis of disaggregated cells. Thus, the FYDR mouse model can be used for studies of mitotic homologous recombination both in vitro and in vivo. PMID- 12750465 TI - The working stroke upon myosin-nucleotide complexes binding to actin. AB - For many years, it has been known that myosin binds to actin tightly, but it had not been possible to devise a muscle fiber experiment to determine whether this binding energy is directly coupled to the working stroke of the actomyosin crossbridge cycle. Addressing the question at the single-molecule level with optical tweezers allows the problem to be resolved. We have compared the working stroke on the binding of four myosin complexes (myosin, myosin-ADP, myosin pyrophosphate, and myosin-adenyl-5'yl imidodiphosphate) with that observed while hydrolyzing ATP. None of the four was observed to give a working stroke significantly different from zero. A working stroke (5.4 nm) was observed only with ATP, which indicates that the other states bind to actin in a rigor-like conformation and that myosin products (M.ADP.Pi), the state that binds to actin during ATPase activity, binds in a different, prestroke conformation. We conclude that myosin, while dissociated from actin, must be able to take up at least two mechanical conformations and show that our results are consistent with these conformations corresponding to the two states characterized at high resolution, which are commonly referred to in terms of having open and closed nucleotide binding pockets. PMID- 12750463 TI - Saccharomyces cerevisiae chromatin-assembly factors that act during DNA replication function in the maintenance of genome stability. AB - Some spontaneous gross chromosomal rearrangements (GCRs) seem to result from DNA replication errors. The chromatin-assembly factor I (CAF-I) and replication coupling assembly factor (RCAF) complexes function in chromatin assembly during DNA replication and repair and could play a role in maintaining genome stability. Inactivation of CAF-I or RCAF increased the rate of accumulating different types of GCRs including translocations and deletion of chromosome arms with associated de novo telomere addition. Inactivation of CAF-I seems to cause damage that activates the DNA-damage checkpoints, whereas inactivation of RCAF seems to cause damage that activates the DNA-damage and replication checkpoints. Both defects result in increased genome instability that is normally suppressed by these checkpoints, RAD52-dependent recombination, and PIF1-dependent inhibition of de novo telomere addition. Treatment of CAF-I- or RCAF-defective cells with methyl methanesulfonate increased the induction of GCRs compared with that seen for a wild-type strain. These results indicate that coupling of chromatin assembly to DNA replication and DNA repair is critical to maintaining genome stability. PMID- 12750466 TI - Evolution of the army ant syndrome: the origin and long-term evolutionary stasis of a complex of behavioral and reproductive adaptations. AB - The army ant syndrome of behavioral and reproductive traits (obligate collective foraging, nomadism, and highly specialized queens) has allowed these organisms to become the premiere social hunters of the tropics, yet we know little about how or why these strategies evolved. The currently accepted view holds that army ants evolved multiple times on separate continents. I generated data from three nuclear genes, a mitochondrial gene, and morphology to test this hypothesis. Results strongly indicate that the suite of behavioral and reproductive adaptations found in army ants throughout the world is inherited from a unique common ancestor, and did not evolve convergently in the New World and Old World as previously thought. New Bayesian methodology for dating the antiquity of lineages by using a combination of fossil and molecular information places the origin of army ants in the mid-Cretaceous, consistent with a Gondwanan origin. Because no known army ant species lacks any component of the army ant syndrome, this group represents an extraordinary case of long-term evolutionary stasis in these adaptations. PMID- 12750467 TI - Targeting Ras signaling through inhibition of carboxyl methylation: an unexpected property of methotrexate. AB - The antifolate methotrexate is one of the most successful drugs in cancer chemotherapy. Although its efficacy is widely attributed to a decrease in nucleotide biosynthesis (1), methotrexate is known to increase homocysteine (2), a compound associated with an elevated risk of heart disease, Alzheimer's disease (3), and neural tube defects (4). A potential mechanism for the detrimental effects of homocysteine is cellular hypomethylation from an increase in S adenosylhomocysteine (5), an inhibitor of methyltransferases including isoprenylcysteine carboxyl methyltransferase (Icmt). Among the substrates of Icmt is the monomeric G protein Ras, a critical component of many signaling pathways that regulate cell growth and differentiation. Because carboxyl methylation of Ras is important for proper plasma membrane localization and function (6), we investigated the role of Icmt in the antiproliferative effect of methotrexate. After methotrexate treatment of DKOB8 cells, Ras methylation is decreased by almost 90%. This hypomethylation is accompanied by a mislocalization of Ras to the cytosol and a 4-fold decrease in the activation of p44 mitogen-activated protein kinase and Akt. Additionally, cells lacking Icmt are highly resistant to methotrexate. Whereas cells expressing wild-type levels of Icmt are inhibited by methotrexate, stable expression of myristoylated H-Ras, which does not require carboxyl methylation for membrane attachment (7), confers resistance to methotrexate. These results suggest that inhibition of Icmt is a critical component of the antiproliferative effect of methotrexate, expanding our understanding of this widely used drug and identifying Icmt as a target for drug discovery. PMID- 12750468 TI - Conservation and molecular dissection of ROUGH SHEATH2 and ASYMMETRIC LEAVES1 function in leaf development. AB - Maize ROUGH SHEATH2 (RS2) and Arabidopsis ASYMMETRIC LEAVES1 (AS1) are orthologous Myb-related genes required for leaf development and act as negative regulators of class 1 KNOTTED1-like homeobox (KNOX) genes in leaf primordia. Expression of RS2 in Arabidopsis fully complements as1 leaf phenotypes and represses the expression of the KNOX gene KNAT1 in leaves. Whereas loss of AS1 function in Arabidopsis results in rounded, lobed leaves with shorter and wider petioles, overexpression of either RS2 or AS1 results in longer and narrower leaves with longer petioles than wild type. A conserved C-terminal domain (CTD) mediates homodimerization of both RS2 and AS1 and modulates leaf shape when expressed independently of the Myb domain in Arabidopsis. Homodimerization is not absolutely required for KNAT1 repression. RS2:GFP fusion protein is biologically active, localized in discrete dynamic subnuclear foci and associates with DNA during cell division. PMID- 12750470 TI - Beyond superquenching: hyper-efficient energy transfer from conjugated polymers to gold nanoparticles. AB - Gold nanoparticles quench the fluorescence of cationic polyfluorene with Stern Volmer constants (KSV) approaching 1011 M-1, several orders of magnitude larger than any previously reported conjugated polymer-quencher pair and 9-10 orders of magnitude larger than small molecule dye-quencher pairs. The dependence of KSV on ionic strength, charge and conjugation length of the polymer, and the dimensions (and thus optical properties) of the nanoparticles suggests that three factors account for this extraordinary efficiency: (i) amplification of the quenching via rapid internal energy or electron transfer, (ii) electrostatic interactions between the cationic polymer and anionic nanoparticles, and (iii) the ability of gold nanoparticles to quench via efficient energy transfer. As a result of this extraordinarily high KSV, quenching can be observed even at subpicomolar concentrations of nanoparticles, suggesting that the combination of conjugated polymers with these nanomaterials can potentially lead to improved sensitivity in optical biosensors. PMID- 12750469 TI - Structure of the coat protein in fd filamentous bacteriophage particles determined by solid-state NMR spectroscopy. AB - The atomic resolution structure of fd coat protein determined by solid-state NMR spectroscopy of magnetically aligned filamentous bacteriophage particles differs from that previously determined by x-ray fiber diffraction. Most notably, the 50 residue protein is not a single curved helix, but rather is a nearly ideal straight helix between residues 7 and 38, where there is a distinct kink, and then a straight helix with a different orientation between residues 39 and 49. Residues 1-5 have been shown to be mobile and unstructured, and proline 6 terminates the helix. The structure of the coat protein in virus particles, in combination with the structure of the membrane-bound form of the same protein in bilayers, also recently determined by solid-state NMR spectroscopy, provides insight into the viral assembly process. In addition to their roles in molecular biology and biotechnology, the filamentous bacteriophages continue to serve as model systems for the development of experimental methods for determining the structures of proteins in biological supramolecular assemblies. New NMR results include the complete sequential assignment of the two-dimensional polarization inversion spin-exchange at the magic angle spectrum of a uniformly 15N-labeled 50 residue protein in a 1.6 x 107 Da particle in solution, and the calculation of the three-dimensional structure of the protein from orientational restraints with an accuracy equivalent to an rms deviation of approximately 1A. PMID- 12750471 TI - Genomewide demarcation of RNA polymerase II transcription units revealed by physical fractionation of chromatin. AB - Epigenetic modifications of chromatin serve an important role in regulating the expression and accessibility of genomic DNA. We report here a genomewide approach for fractionating yeast chromatin into two functionally distinct parts, one containing RNA polymerase II transcribed sequences, and the other comprising noncoding sequences and genes transcribed by RNA polymerases I and III. Noncoding regions could be further fractionated into promoters and segments lacking promoters. The observed separations were apparently based on differential crosslinking efficiency of chromatin in different genomic regions. The results reveal a genomewide molecular mechanism for marking promoters and genomic regions that have a license to be transcribed by RNA polymerase II, a previously unrecognized level of genomic complexity that may exist in all eukaryotes. Our approach has broad potential use as a tool for genome annotation and for the characterization of global changes in chromatin structure that accompany different genetic, environmental, and disease states. PMID- 12750472 TI - Beta cell MHC class I is a late requirement for diabetes. AB - Type 1 diabetes occurs as a result of an autoimmune attack on the insulin producing beta cells. Although CD8 T cells have been implicated both early and late in this process, the requirement for direct interaction between these cells and MHC class I on the beta cells has not been demonstrated. By using nonobese diabetic mice lacking beta cell class I expression, we show that both initiation and progression of insulitis proceeds unperturbed. However, without beta cell class I expression, the vast majority of these mice do not develop hyperglycemia. These findings demonstrate that a direct interaction between CD8 T cells and beta cells is not required for initiation or early disease progression. The requirement for class I on beta cells is a relatively late checkpoint in the development of diabetes. PMID- 12750473 TI - How the BfiI restriction enzyme uses one active site to cut two DNA strands. AB - Unlike other restriction enzymes, BfiI functions without metal ions. It recognizes an asymmetric DNA sequence, 5'-ACTGGG-3', and cuts top and bottom strands at fixed positions downstream of this sequence. Many restriction enzymes are dimers of identical subunits, with one active site for each DNA strand. Others, like FokI, dimerize transiently during catalysis. BfiI is also a dimer but it has only one active site, at the dimer interface. We show here that BfiI remains a dimer as it makes double-strand breaks in DNA and that its single active site acts sequentially, first on the bottom and then the top strand. Hence, after cutting the bottom strand, a rearrangement of either the protein and/or the DNA in the BfiI-DNA complex must switch the active site to the top strand. Low pH values selectively block top-strand cleavage, converting BfiI into a nicking enzyme that cleaves only the bottom strand. The switch to the top strand may depend on the ionization of the cleaved 5' phosphate in the bottom strand. BfiI thus uses a mechanism for making double-strand breaks that is novel among restriction enzymes. PMID- 12750474 TI - Global perception in small brains: topological pattern recognition in honey bees. AB - A series of experiments with honey bees demonstrate that their small brains nevertheless possess the ability for topological perception. Bees rapidly learned to discriminate patterns that are topologically different, and they generalized the learned cue to other novel patterns. By contrast, discrimination of topologically equivalent patterns was learned much more slowly and not as well. Thus, although the global nature of topological properties makes their computation difficult, topology may be a fundamental component of the vocabulary by which visual systems represent and characterize objects. PMID- 12750475 TI - sal1 determines the number of aleurone cell layers in maize endosperm and encodes a class E vacuolar sorting protein. AB - A microscopy-based screen of a large collection of maize Mutator (Mu) transposon lines identified the supernumerary aleurone layers 1-1 (sal1-1) mutant line carrying up to seven layers of aleurone cells in defective kernel endosperm compared with only a single layer in wild-type grains. Normal, well filled endosperm that is homozygous for the sal1-1 mutant allele contains two to three layers of aleurone cells. Cloning of the sal1 gene was accomplished by using Mu tagging, and the identity of the cloned gene was confirmed by isolating an independent sal1-2 allele by reverse genetics. Homozygous sal1-2 endosperm has two to three layers of aleurone cells in normal, well filled grains. In situ hybridization experiments reveal that the sal1 gene is ubiquitously expressed in vegetative as well as zygotic grain tissues, with no difference being detected between aleurone cells and starchy endosperm cells. Northern blot analysis failed to detect the sal1-2 transcript in leaves of homozygous plants, suggesting that the allele is a true sal1 knockout allele. The sal1 gene encodes a homologue of the human Chmp1 gene, a member of the conserved family of the class E vacuolar protein sorting genes implicated in membrane vesicle trafficking. In mammals, CHMP1 functions in the pathway targeting plasma membrane receptors and ligands to lysosomes for proteolytic degradation. Possible roles for the function of the sal1 gene in aleurone signaling, including a defect in endosome trafficking, are discussed. PMID- 12750476 TI - A new Drosophila spliceosomal intron position is common in plants. AB - The 25-year-old debate about the origin of introns between proponents of "introns early" and "introns late" has yielded significant advances, yet important questions remain to be ascertained. One question concerns the density of introns in the last common ancestor of the three multicellular kingdoms. Approaches to this issue thus far have relied on counts of the numbers of identical intron positions across present-day taxa on the assumption that the introns at those sites are orthologous. However, dismissing parallel intron gain for those sites may be unwarranted, because various factors can potentially constrain the site of intron insertion. Demonstrating parallel intron gain is severely handicapped, because intron sequences often evolve exceedingly fast and intron phylogenetic distributions are usually ambiguous, such that alternative loss and gain scenarios cannot be clearly distinguished. We have identified an intron position that was gained independently in animals and plants in the xanthine dehydrogenase gene. The extremely disjointed phylogenetic distribution of the intron argues strongly for separate gain rather than recurrent loss. If the observed phylogenetic pattern had resulted from recurrent loss, all observational support previously gathered for the introns-late theory of intron origins based on the phylogenetic distribution of introns would be invalidated. PMID- 12750477 TI - Cytokines and BCR-ABL mediate suppression of TRAIL-induced apoptosis through inhibition of forkhead FOXO3a transcription factor. AB - Cytokine-provided survival signals are known to suppress apoptosis through inhibition of mitochondrial pathways that involve Bcl-2 family members. Here we show that in hematopoietic cells, cytokines also regulate death receptor-mediated pathways. We demonstrate that hematopoietic cytokines such as IL-3 and erythropoietin in normal cells, as well as BCR-ABL oncoprotein in transformed cells, inhibit transcription of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). Using small interfering RNAs, we show that the inhibition of TRAIL function is sufficient to partially rescue cytokine-deprived cells from apoptosis. Finally, we demonstrate that cytokine and BCR-ABL suppression of TRAIL transcription is mediated through phosphorylation and inhibition of the forkhead FOXO3a transcription factor. BCR-ABL-induced inhibition of TRAIL transcription in hematopoietic cells may provide a novel mechanism for tumorigenicity in chronic myeloid leukemia. PMID- 12750479 TI - Entangled macroscopic quantum States in two superconducting qubits. AB - We present spectroscopic evidence for the creation of entangled macroscopic quantum states in two current-biased Josephson-junction qubits coupled by a capacitor. The individual junction bias currents are used to control the interaction between the qubits by tuning the energy level spacings of the junctions in and out of resonance with each other. Microwave spectroscopy in the 4 to 6 gigahertzrange at 20 millikelvin reveals energy levels that agree well with theoretical results for entangled states. The single qubits are spatially separate, and the entangled states extend over the 0.7-millimeter distance between the two qubits. PMID- 12750478 TI - Translation of polarity cues into asymmetric spindle positioning in Caenorhabditis elegans embryos. AB - Asymmetric divisions are crucial for generating cell diversity; they rely on coupling between polarity cues and spindle positioning, but how this coupling is achieved is poorly understood. In one-cell stage Caenorhabditis elegans embryos, polarity cues set by the PAR proteins mediate asymmetric spindle positioning by governing an imbalance of net pulling forces acting on spindle poles. We found that the GoLoco-containing proteins GPR-1 and GPR-2, as well as the Galpha subunits GOA-1 and GPA-16, were essential for generation of proper pulling forces. GPR-1/2 interacted with guanosine diphosphate-bound GOA-1 and were enriched on the posterior cortex in a par-3- and par-2-dependent manner. Thus, the extent of net pulling forces may depend on cortical Galpha activity, which is regulated by anterior-posterior polarity cues through GPR-1/2. PMID- 12750480 TI - Using 1-Hz GPS data to measure deformations caused by the Denali fault earthquake. AB - The 3 November 2002 moment magnitude 7.9 Denali fault earthquake generated large, permanent surface displacements in Alaska and large-amplitude surface waves throughout western North America. We find good agreement between strong ground motion records integrated to displacement and 1-hertz Global Positioning System (GPS) position estimates collected approximately 140 kilometers from the earthquake epicenter. One-hertz GPS receivers also detected seismic surface waves 750 to 3800 kilometers from the epicenter, whereas these waves saturated many of the seismic instruments in the same region. High-frequency GPS increases the dynamic range and frequency bandwidth of ground-motion observations, providing another tool for studying earthquake processes. PMID- 12750481 TI - AIDS and Africa: still a sad story. PMID- 12750482 TI - SARS outbreak. Flood of sequence data yields clues but few answers. PMID- 12750483 TI - SARS outbreak. Hong Kong to beef up monitoring. PMID- 12750484 TI - Physics. Is Berkeley past its prime? PMID- 12750485 TI - Anthrax vaccine. NIAID's $233 million problem put on hold. PMID- 12750486 TI - Toxicology. Arsenic victims to take British science body to court. PMID- 12750487 TI - Number theory. Prime-number proof's leap falls short. PMID- 12750488 TI - Paleontology. Life's diversity may truly have leaped since the dinosaurs. PMID- 12750489 TI - Toxicology. E.U. shifts endocrine disrupter research into overdrive. PMID- 12750490 TI - Scientific workforce. Down for the count? PMID- 12750492 TI - Scientific workforce. Report to argue for more training funds. PMID- 12750493 TI - Taiwan-China collaboration. A bridge over troubled waters. PMID- 12750491 TI - Scientific workforce. Viewed from 1986, it's mostly downhill. PMID- 12750495 TI - Particle physics. Monster machines. PMID- 12750494 TI - Particle physics. Calculating the incalculable. PMID- 12750496 TI - Cognition. How the mind reads other minds. PMID- 12750497 TI - Weather forecasting. Huge Pacific waves trigger wild weather half a world away. PMID- 12750498 TI - Eating disorders and obesity. PMID- 12750500 TI - The "big picture" in obesity research. PMID- 12750499 TI - Confusing food with obesity. PMID- 12750501 TI - Building biosafety laboratories. PMID- 12750502 TI - Another epidemic of politics? PMID- 12750503 TI - Exploring sea-floor resources. PMID- 12750504 TI - Disaster management. Flood risk in Central Europe. PMID- 12750505 TI - Immunology. A viral on/off switch for interferon. PMID- 12750506 TI - Molecular biology. SMC complexes--wrapped up in controversy. PMID- 12750507 TI - Atmospheric science. Climate forcing by aerosol--a hazy picture. PMID- 12750508 TI - Materials science. Molecular fuel tanks. PMID- 12750509 TI - Geophysics. How faults accommodate plate motion. PMID- 12750510 TI - Applications for nuclear phenomena generated by ultra-intense lasers. AB - The amplification of laser light to generate powers large enough to affect the nucleus has been the desire of scientists since the invention of the laser 40 years ago. Many lasers, including tabletop varieties, now have pulse powers greater than the electrical power generated by all the world's power plants combined. When this power is focused to dimensions of a few microns, laser-driven nuclear phenomena can occur. Here we review the developments in this research field and describe the potential of laser produced proton, neutron, and heavy ion beams, together with isotope and isomer production. PMID- 12750512 TI - The 2002 Denali fault earthquake, Alaska: a large magnitude, slip-partitioned event. AB - The MW (moment magnitude) 7.9 Denali fault earthquake on 3 November 2002 was associated with 340 kilometers of surface rupture and was the largest strike-slip earthquake in North America in almost 150 years. It illuminates earthquake mechanics and hazards of large strike-slip faults. It began with thrusting on the previously unrecognized Susitna Glacier fault, continued with right-slip on the Denali fault, then took a right step and continued with right-slip on the Totschunda fault. There is good correlation between geologically observed and geophysically inferred moment release. The earthquake produced unusually strong distal effects in the rupture propagation direction, including triggered seismicity. PMID- 12750511 TI - Hypermutation of HIV-1 DNA in the absence of the Vif protein. PMID- 12750513 TI - Slip partitioning by elastoplastic propagation of oblique slip at depth. AB - Oblique motion along tectonic boundaries is commonly partitioned into slip on faults with different senses of motion. The origin of slip partitioning is important to structural geology, tectonophysics, and earthquake mechanics. Partitioning can be explained by the upward elastoplastic propagation of oblique slip from a fault or shear zone at depth. The strain field ahead of the propagating fault separates into zones of predominantly normal, reverse, and strike-slip faulting. The model successfully predicts the distribution of fault types along parts of the San Andreas and Haiyuan faults. PMID- 12750515 TI - Hydrogen storage in microporous metal-organic frameworks. AB - Metal-organic framework-5 (MOF-5) of composition Zn4O(BDC)3 (BDC = 1,4 benzenedicarboxylate) with a cubic three-dimensional extended porous structure adsorbed hydrogen up to 4.5 weight percent (17.2 hydrogen molecules per formula unit) at 78 kelvin and 1.0 weight percent at room temperature and pressure of 20 bar. Inelastic neutron scattering spectroscopy of the rotational transitions of the adsorbed hydrogen molecules indicates the presence of two well-defined binding sites (termed I and II), which we associate with hydrogen binding to zinc and the BDC linker, respectively. Preliminary studies on topologically similar isoreticular metal-organic framework-6 and -8 (IRMOF-6 and -8) having cyclobutylbenzene and naphthalene linkers, respectively, gave approximately double and quadruple (2.0 weight percent) the uptake found for MOF-5 at room temperature and 10 bar. PMID- 12750514 TI - Detection of hydroxyl ions in bone mineral by solid-state NMR spectroscopy. AB - Previous measurements of the hydroxyl (OH-) ion content of the calcium phosphate crystals of bone mineral have indicated a substantial depletion or near-absence of OH-, despite its presumed status as a constituent of the hydroxyapatite lattice. Analytical methods for determining bone crystal OH- content have depended on procedures or assumptions that may have biased the results, such as chemical pretreatment to eliminate interference from the organic matrix. We demonstrate a two-dimensional solid-state nuclear magnetic resonance (NMR) spectroscopy technique that detects the proton spectrum of bone crystals while suppressing the interfering matrix signals, eliminating the need for specimen pretreatment other than cryogenic grinding. Results on fresh-frozen and ground whole bone of several mammalian species show that the bone crystal OH- is readily detectable; a rough estimate yields an OH- content of human cortical bone of about 20% of the amount expected in stoichiometric hydroxyapatite. This finding sheds light on the biochemical processes underlying normal and abnormal bone mineral metabolism. PMID- 12750516 TI - Giant magnetic anisotropy of single cobalt atoms and nanoparticles. AB - The isotropic magnetic moment of a free atom is shown to develop giant magnetic anisotropy energy due to symmetry reduction at an atomically ordered surface. Single cobalt atoms deposited onto platinum (111) are found to have a magnetic anisotropy energy of 9 millielectron volts per atom arising from the combination of unquenched orbital moments (1.1 Bohr magnetons) and strong spin-orbit coupling induced by the platinum substrate. By assembling cobalt nanoparticles containing up to 40 atoms, the magnetic anisotropy energy is further shown to be dependent on single-atom coordination changes. These results confirm theoretical predictions and are of fundamental value to understanding how magnetic anisotropy develops in finite-sized magnetic particles. PMID- 12750517 TI - The impact of the pull of the recent on the history of marine diversity. AB - Up to 50% of the increase in marine animal biodiversity through the Cenozoic at the genus level has been attributed to a sampling bias termed "the Pull of the Recent," the extension of stratigraphic ranges of fossil taxa by the relatively complete sampling of the Recent biota. However, 906 of 958 living genera and subgenera of bivalve mollusks having a fossil record occur in the Pliocene or Pleistocene. The Pull of the Recent thus accounts for only 5% of the Cenozoic increase in bivalve diversity, a major component of the marine record, suggesting that the diversity increase is likely to be a genuine biological pattern. PMID- 12750518 TI - Local selection and latitudinal variation in a marine predator-prey interaction. AB - Although pairs of species often interact over broad geographic ranges, few studies have explored how interactions vary across these large spatial scales. Surveys along 1500 kilometers of the Pacific coast of North America documented marked variation in the frequency of predation by the snail Nucella canaliculata on the intertidal mussel Mytilus californianus. Laboratory rearing experiments suggest that regional differences in drilling behavior have a genetic basis, and mitochondrial sequence variation confirms that gene flow is low among these snail populations. Marine communities separated by hundreds of kilometers may have intrinsically different dynamics, with interactions shaped by restricted gene flow and spatially varying selection. PMID- 12750519 TI - Rapid turnover of hyphae of mycorrhizal fungi determined by AMS microanalysis of 14C. AB - Processes in the soil remain among the least well-characterized components of the carbon cycle. Arbuscular mycorrhizal (AM) fungi are ubiquitous root symbionts in many terrestrial ecosystems and account for a large fraction of photosynthate in a wide range of ecosystems; they therefore play a key role in the terrestrial carbon cycle. A large part of the fungal mycelium is outside the root (the extraradical mycelium, ERM) and, because of the dispersed growth pattern and the small diameter of the hyphae (<5 micrometers), exceptionally difficult to study quantitatively. Critically, the longevity of these fine hyphae has never been measured, although it is assumed to be short. To quantify carbon turnover in these hyphae, we exposed mycorrhizal plants to fossil ("carbon-14-dead") carbon dioxide and collected samples of ERM hyphae (up to 116 micrograms) over the following 29 days. Analyses of their carbon-14 content by accelerator mass spectrometry (AMS) showed that most ERM hyphae of AM fungi live, on average, 5 to 6 days. This high turnover rate reveals a large and rapid mycorrhizal pathway of carbon in the soil carbon cycle. PMID- 12750520 TI - Mitochondrial dysfunction in the elderly: possible role in insulin resistance. AB - Insulin resistance is a major factor in the pathogenesis of type 2 diabetes in the elderly. To investigate how insulin resistance arises, we studied healthy, lean, elderly and young participants matched for lean body mass and fat mass. Elderly study participants were markedly insulin-resistant as compared with young controls, and this resistance was attributable to reduced insulin-stimulated muscle glucose metabolism. These changes were associated with increased fat accumulation in muscle and liver tissue assessed by 1H nuclear magnetic resonance (NMR) spectroscopy, and with a approximately 40% reduction in mitochondrial oxidative and phosphorylation activity, as assessed by in vivo 13C/31P NMR spectroscopy. These data support the hypothesis that an age-associated decline in mitochondrial function contributes to insulin resistance in the elderly. PMID- 12750521 TI - Regulation of aging and age-related disease by DAF-16 and heat-shock factor. AB - The Caenorhabditis elegans transcription factor HSF-1, which regulates the heat shock response, also influences aging. Reducing hsf-1 activity accelerates tissue aging and shortens life-span, and we show that hsf-1 overexpression extends lifespan. We find that HSF-1, like the transcription factor DAF-16, is required for daf-2-insulin/IGF-1 receptor mutations to extend life-span. Our findings suggest this is because HSF-1 and DAF-16 together activate expression of specific genes, including genes encoding small heat-shock proteins, which in turn promote longevity. The small heat-shock proteins also delay the onset of polyglutamine expansion protein aggregation, suggesting that these proteins couple the normal aging process to this type of age-related disease. PMID- 12750522 TI - Distinct cohesin complexes organize meiotic chromosome domains. AB - Meiotic cohesin complexes at centromeres behave differently from those along chromosome arms, but the basis for these differences has remained elusive. The fission yeast cohesin molecule Rec8 largely replaces its mitotic counterpart, Rad21/Scc1, along the entire chromosome during meiosis. Here we show that Rec8 complexes along chromosome arms contain Rec11, whereas those in the vicinity of centromeres have a different partner subunit, Psc3. The arm associated Rec8-Rec11 complexes are critical for meiotic recombination. The Rec8-Psc3 complexes comprise two different types of assemblies. First, pericentromeric Rec8-Psc3 complexes depend on histone methylation-directed heterochromatin for their localization and are required for cohesion during meiosis II. Second, central core Rec8-Psc3 complexes form independently of heterochromatin and are presumably required for establishing monopolar attachment at meiosis I. These findings define distinct modes of assembly and functions for cohesin complexes at different regions along chromosomes. PMID- 12750523 TI - Tumor response to radiotherapy regulated by endothelial cell apoptosis. AB - About 50% of cancer patients receive radiation therapy. Here we investigated the hypothesis that tumor response to radiation is determined not only by tumor cell phenotype but also by microvascular sensitivity. MCA/129 fibrosarcomas and B16F1 melanomas grown in apoptosis-resistant acid sphingomyelinase (asmase)-deficient or Bax-deficient mice displayed markedly reduced baseline microvascular endothelial apoptosis and grew 200 to 400% faster than tumors on wild-type microvasculature. Thus, endothelial apoptosis is a homeostatic factor regulating angiogenesis-dependent tumor growth. Moreover, these tumors exhibited reduced endothelial apoptosis upon irradiation and, unlike tumors in wild-type mice, they were resistant to single-dose radiation up to 20 grays (Gy). These studies indicate that microvascular damage regulates tumor cell response to radiation at the clinically relevant dose range. PMID- 12750524 TI - Catalysis of ribosomal translocation by sparsomycin. AB - During protein synthesis, transfer RNAs (tRNAs) are translocated from the aminoacyl to peptidyl to exit sites of the ribosome, coupled to the movement of messenger RNA (mRNA), in a reaction catalyzed by elongation factor G (EF-G) and guanosine triphosphate (GTP). Here, we show that the peptidyl transferase inhibitor sparsomycin triggers accurate translocation in vitro in the absence of EF-G and GTP. Our results provide evidence that translocation is a function inherent to the ribosome and that the energy to drive this process is stored in the tRNA-mRNA-ribosome complex after peptide-bond formation. These findings directly implicate the peptidyl transferase center of the 50S subunit in the mechanism of translocation, a process involving large-scale movement of tRNA and mRNA in the 30S subunit, some 70 angstroms away. PMID- 12750525 TI - Editorial comment: How appropriate is carotid endarterectomy? PMID- 12750526 TI - Editorial comment: Protein-energy undernutrition and acute stroke outcome. PMID- 12750527 TI - "Mobile stroke unit" for hyperacute stroke treatment. PMID- 12750528 TI - Dysregulation of the levels of matrix metalloproteinases and tissue inhibitors of matrix metalloproteinases in the early phase of cerebral ischemia. PMID- 12750529 TI - Nonsteroidal anti-inflammatory drugs as risk factors for spontaneous intracerebral hemorrhage and aneurysmal subarachnoid hemorrhage. PMID- 12750530 TI - Symptomatic carotid artery stenosis: heterogeneity of destabilizing mechanisms? PMID- 12750531 TI - Carotid angioplasty with stenting and carotid endarterectomy for high-risk patients. PMID- 12750532 TI - Evacuation of intracerebral hematoma is likely to be beneficial--against. PMID- 12750533 TI - Evacuation of intracerebral hematoma is likely to be beneficial. PMID- 12750534 TI - Surgery for intracerebral hemorrhage: an evidence-poor zone. PMID- 12750535 TI - Trends in stroke incidence and 10-year survival in Soderhamn, Sweden, 1975-2001. AB - BACKGROUND AND PURPOSE: Stroke mortality rates have declined in many countries. Stroke incidence rates have also declined, but not to the same extent and not always in parallel with stroke mortality. The aim of the present study was to investigate trends in stroke incidence and long-term survival in a Swedish population. METHODS: A population-based survey of the incidence of first-ever stroke was performed prospectively in the periods 1975 to 1977, 1983 to 1986, and 1987 to 1990. Case fatality ratios and survival rates were followed to 2001. RESULTS: Crude incidence rates increased between 1975 to 1977 and 1983 to 1986, but age- and sex-adjusted incidence rates were stable during the whole period of 1975 to 1990. The incidence of intracerebral hemorrhage decreased by approximately two thirds, whereas the incidence of mild brain infarction almost doubled. The case fatality ratio at 28 days did not change, but the 10-year survival ratio increased from 13% to 35%. Mean survival time increased significantly among patients with intracerebral hemorrhage and brain infarction but not among patients with subarachnoidal hemorrhage or stroke of undetermined origin. CONCLUSIONS: Stroke incidence and short-term case fatality did not change between 1975 and 1990 in the Soderhamn population. Long-term survival after stroke, on the other hand, has continued to improve to 2001. The implication of these changes is that the burden of stroke is likely to increase unless strokes are becoming less severe. PMID- 12750536 TI - Poor nutritional status on admission predicts poor outcomes after stroke: observational data from the FOOD trial. AB - BACKGROUND AND PURPOSE: Previous studies suggest that undernourished patients with acute stroke do badly. The data, however, are not robust. We aimed to reliably assess the importance of baseline nutritional status as an independent predictor of long-term outcome after stroke in a large prospective cohort enrolled in the Feed Or Ordinary Diet (FOOD) trial, a multicenter randomized trial evaluating various feeding policies. METHODS: Patients admitted to hospital with a recent stroke were enrolled in the FOOD trial. Data on nutritional status and other clinical predictors of outcome were collected at trial entry. At 6 months, the coordinating center collected data on survival and functional status (modified Rankin Scale). Outcome assessment was done by researchers blinded to baseline assessments and treatment allocation. RESULTS: Between November 1996 and November 2001, 3012 patients were enrolled, and 2955 (98%) were followed up. Of the 275 undernourished patients, 102 (37%) were dead by final follow-up compared with only 445 (20%) of 2194 patients of normal nutritional status (odds ratio [OR], 2.32; 95% CI, 1.78 to 3.02). After adjustment for age, prestroke functional state, and stroke severity, this relationship, although weakened, still held (OR, 1.82; 95% CI, 1.34 to 2.47). Undernourished patients were more likely to develop pneumonia, other infections, and gastrointestinal bleeding during their hospital admission than other patients. CONCLUSIONS: These data provide reliable evidence that nutritional status early after stroke is independently associated with long term outcome. It supports the rationale for the FOOD trial, which continues to recruit and aims to estimate the effect of different feeding regimes on outcome after stroke and thus determine whether the association observed in this study is likely to be causal. PMID- 12750537 TI - Acute focal neurological deficits in aneurysmal subarachnoid hemorrhage: relation of clinical course, CT findings, and metabolite abnormalities monitored with bedside microdialysis. AB - BACKGROUND AND PURPOSE: We sought (1) to identify early metabolic markers for the development of (ir)reversible neurological deficits and cerebral infarction in subarachnoid hemorrhage (SAH) patients by using the microdialysis technique and (2) to evaluate the influence of intracerebral hemorrhage (ICH) on microdialysis parameters. METHODS: We performed a prospective study of 44 SAH patients with acute focal neurological deficits (AFND) occurring acutely with SAH (due to ICH) or directly after surgery (due to clip stenosis, thromboembolism, or early edema). Fifty-one nonischemic SAH patients served as a control group. A microdialysis catheter was inserted into the vascular territory of the aneurysm after clipping. The microdialysates were analyzed hourly for extracellular glucose, lactate, lactate/pyruvate ratio, glutamate, and glycerol with a bedside analyzer. Microdialysis-related CT findings were evaluated for the presence of ICH and cerebral infarction. Reversibility of neurological symptoms after 4 weeks and 6- and 12-month outcomes were assessed. RESULTS: In patients with AFND, cerebral metabolism was severely disturbed when microdialysis started compared with controls (P<0.005). Infarction on CT was associated with pathological microdialysis parameters (P<0.002) and development of a fixed deficit (P<0.003), while the presence of ICH alone was not. A secondary neurological deterioration of AFND patients (n=11) was reflected by preceding (0 to 20 hours) changes of microdialysate concentrations. CONCLUSIONS: In the presence of ICH, pathological microdialysis values may indicate reversible tissue damage. Extreme microdialysis values and pathological microdialysis concentrations that further deteriorate 2 fold are highly indicative of the development of cerebral infarction and permanent neurological deficits. Therefore, the analysis of relative changes of microdialysis parameters is crucial for the detection of ischemia in SAH patients. PMID- 12750538 TI - Homocysteine and cerebral infarction in finnish male smokers. AB - BACKGROUND AND PURPOSE: Homocysteine is associated with stroke, but it is not clear whether this relationship is causal. We examined the association between total serum homocysteine concentration (tHcy) and cerebral infarction in a cohort of Finnish male smokers. METHODS: This is a matched case-control study of 201 cases of cerebral infarction and 201 concurrently sampled age-matched controls nested in a cohort of 13 840 male smokers free of cardiovascular disease at the completion of the Alpha-Tocopherol and Beta-Carotene (ATBC) Cancer Prevention study. Conditional logistic regression was used to calculate odds ratios (ORs) and to adjust for confounding variables. An unmatched analysis was also performed. RESULTS: The geometric mean tHcy was 13.3 micromol/L (95% CI, 12.6 to 13.9) in cases and 12.6 micromol/L (95% CI, 12.0 to 13.2) in controls (P=0.09). There was a graded increase in the OR of cerebral infarction per quartile increase in tHcy (OR, 1.0, 1.7, 1.9, 2.1; trend P=0.02; 201 case-control pairs) when adjusted for traditional risk factors. There was a similar trend in a subgroup of 120 case-control pairs for which further adjustment for lifestyle factors was possible (OR, 1.0, 1.9, 2.5, 2.2; trend P=0.07 in the matched analyses; OR, 1.0, 1.2, 1.9, 2.0; trend P=0.02 in the unmatched analyses). The adjusted OR per 1-SD increase in log-transformed tHcy (equivalent to 4.7 micromol) was 1.4 (95% CI, 1.1 to 1.7; P=0.01). CONCLUSIONS: tHcy appears to predict cerebral infarction in Finnish male smokers. PMID- 12750539 TI - Neurological dysfunctions versus regional infarction volume after focal ischemia in Mongolian gerbils. AB - BACKGROUND AND PURPOSE: With advances in the therapy of stroke at the postacute phase, the use of animal models for chronological and region-specific evaluation of neurological function has become increasingly important. Our aim was to test long-term behavioral dysfunction in gerbils after focal ischemia and to correlate the results with the regional distribution of infarction in the coordinating cortical regions. METHODS: Repetitive unilateral hemispheric ischemia (two 10 minute occlusions, 5-hour interval) was induced in Mongolian gerbils. The elevated body swing test (EBST), bilateral asymmetry test (BAT), and T-maze test were performed to assess asymmetrical motor behavior, somatosensory deficit, and spatial cognitive dysfunction during 4 weeks after ischemia. The results were correlated against the regional infarction volume of the primary motor, somatosensory, and primary visual cortices at 4 weeks after ischemia. RESULTS: In all postischemic gerbils, persistent sensorimotor and cognitive dysfunctions were detectable throughout the postischemic period. Histological examination revealed that a cortical zone of infarction surrounded the selective neuronal death in the ipsilateral cerebral hemisphere. The regional infarction volumes of the primary motor, somatosensory, and visual cortices were significantly correlated with the scores of the EBST, BAT, and T-maze test, respectively. These combinations had the highest regression coefficient of all pairs. CONCLUSIONS: Postischemic motor and somatosensory functions were significantly correlated with regional infarction volumes in the corresponding cortical regions. In gerbils, visual abnormality could be independently detected by the T-maze test. Such regional analyses of ischemic lesions would be useful for investigating the functional outcomes of stroke therapy. PMID- 12750540 TI - Increase in plasma matrix metalloproteinase-9 in acute stroke patients with thrombolysis failure. AB - BACKGROUND AND PURPOSE: Platelet-rich thrombi are resistant to thrombolytics. Matrix metalloproteinases (MMPs) may be involved in platelet aggregation and contribute to thrombolysis failure in stroke patients. METHODS: Plasma samples from 23 stroke patients who had received thrombolytics and from 47 healthy volunteers were examined for MMP-2 and MMP-9 by both enzyme-linked immunosorbent assays and zymography. RESULTS: The arteries were recanalized in 15 patients but not in the other 8. The MMP-9 plasma level was significantly higher in patients whose arteries were not recanalized. CONCLUSIONS: MMP-9 may be associated with the formation of a thrombolytics-resistant thrombus. PMID- 12750541 TI - Alterations in autoregulatory and myogenic function in the cerebrovasculature of Dahl salt-sensitive rats. AB - BACKGROUND AND PURPOSE: Dahl salt-sensitive rats fed an 8.7% NaCl diet exhibited hypertensive encephalopathy and developed seizures associated with areas of blood brain barrier (BBB) disruption without brain ischemia. The incidence of hemorrhagic stroke was low (7/47). We tested the hypothesis that a defect in cerebral blood flow (CBF) autoregulation under hypertensive conditions preceded hypertensive encephalopathy. METHODS: Brain ischemia and BBB disruption were assessed with the use of tetrazolium red staining and Evans blue dye extravasation, respectively. Myogenic constriction to pressure was measured in isolated middle cerebral arteries (MCAs) with a pressure myograph. CBF autoregulation was assessed with the use of laser-Doppler techniques. RESULTS: Asymptomatic rats fed 8.7% NaCl had MCAs that developed an age-related attenuation in their ability to constrict to pressure, which was amplified in rats exhibiting hypertensive encephalopathy. The MCAs of rats with hemorrhagic stroke lost this function and developed large degrees of basal tone. The majority (4/6) of asymptomatic rats fed high salt for longer than 3 weeks exhibited a linear relationship between CBF and blood pressure. The characteristics of CBF regulation were consistent with the possible absence of autoregulation coupled with cerebrovascular vasoconstriction. CONCLUSIONS: Both MCA pressure-dependent constriction and CBF autoregulation in the MCA perfusion domain were lost before the development of hypertensive encephalopathy or hemorrhagic stroke. These defects could contribute to the development of BBB disruption during hypertension. Cerebrovascular vasoconstriction in the absence of CBF autoregulation may protect the brain from excessive overperfusion during hypertension and could account for the low incidence of cerebral hemorrhage in this model. PMID- 12750542 TI - Stroke development in stroke-prone spontaneously hypertensive rats alters the ability of cerebrovascular muscle to utilize internal Ca2+ to elicit constriction. AB - BACKGROUND AND PURPOSE: The ability of middle cerebral arteries (MCAs) to utilize intracellular smooth muscle (SM) Ca2+ to produce constriction in response to pressure and agonists was assessed in relation to hemorrhagic stroke development in Wistar-Kyoto stroke-prone (SHRSP) and stroke-resistant (srSHR) spontaneously hypertensive rats. METHODS: MCAs were studied with the use of a pressure myograph at 100 mm Hg. RESULTS: MCAs from srSHR and prestroke SHRSP exhibited pressure dependent constriction and constricted in response to vasopressin or serotonin in the presence of nifedipine or the absence of [Ca2+]o. MCAs from poststroke SHRSP lost the latter functions and could only constrict in response to vasopressin/serotonin in Krebs' solution containing Ca2+ in the absence of nifedipine. This indicated that the SM could not utilize internal Ca2+ for constriction and maintained constriction by Ca2+ entry through L-type channels. The MCAs of poststroke SHRSP could not constrict to [K+]o-induced depolarization, suggesting that the agonist-induced opening of the L-type channels occurred by mechanisms other than SM depolarization. Depletion of the sarcoplasmic SM Ca2+ stores of MCAs from srSHR with cyclopiazonic acid did not prevent pressure dependent constriction. CONCLUSIONS: Stroke in SHRSP produced a defect in the ability of MCAs to constrict in response to vasopressin or serotonin via the use of an intracellular source of Ca2+. This could be promoted by an inability of the SM to release intracellular Ca2+, by the depletion of internal Ca2+ stores, or by a decrease in the contractile sensitivity to Ca2+ released from the internal stores. PMID- 12750543 TI - Impact of establishing a primary stroke center at a community hospital on the use of thrombolytic therapy: the NINDS Suburban Hospital Stroke Center experience. AB - BACKGROUND AND PURPOSE: To increase the proportion of ischemic stroke patients treated with thrombolytic therapy, the establishment of primary stroke centers in community hospitals has been advocated. We evaluated the use of thrombolytic therapy before and after institution of a primary stroke center in a community hospital. METHODS: The availability of an on-call stroke emergency response team was the only significant additional resource required for this hospital. All eligible patients were treated with intravenous tissue plasminogen activator (tPA). The number of patients with cerebrovascular disease, number and proportion of patients treated with tPA, times to treatment, and patient outcomes were recorded during the first 2 years of the stroke center. RESULTS: During the 12 months before institution of the stroke center, 3 ischemic stroke patients (1.5%) were treated with tPA. During the 2-year period of around-the-clock coverage, 44 of 420 ischemic stroke patients (10.5%) were treated with intravenous tPA, a significant increase in tPA use (P<0.0001). CONCLUSIONS: Establishment of a primary stroke center at a community hospital resulted in a substantial increase in the proportion of patients receiving thrombolytic therapy for ischemic stroke. If this experience is generalized, the beneficial impact of primary stroke centers on stroke outcomes and costs to the healthcare system may be substantial. PMID- 12750544 TI - Cause of stroke recurrence is multifactorial: patterns, risk factors, and outcomes of stroke recurrence in the South London Stroke Register. AB - BACKGROUND AND PURPOSE: This article examines stroke recurrence and whether the subtype of the initial stroke influences the risk and subtypes of further strokes. The proportion of recurrences attributable to conventional risk factors is quantified. METHODS: From January 1995 to August 2000, all first-in-a-lifetime strokes (n=1626) were identified and prospectively followed up in a defined multiethnic inner city population of 234 533. Twelve overlapping referral sources and face-to-face follow-up at 3 months and 1 and 3 years were used to attain complete registration of stroke recurrence. Index and recurrent stroke were classified according to the Oxford Community Stroke Project classification. RESULTS: In 2744 person-years of follow-up, 153 recurrences were observed. At 5 years, the cumulative risk of first stroke recurrence was 16.6% (95% CI, 13.5 to 20.4), and the combined risk of death or stroke recurrence was 65.3% (95% CI, 61.9 to 68.6). Ethnicity and subtype of index stroke were not associated with stroke recurrence. A change in subtype between index and recurrent stroke occurred in 45.5% (95% CI, 35.8 to 55.2) of cases and was most frequent among index lacunar strokes and primary intracerebral hemorrhages. In multivariable analyses, diabetes mellitus and atrial fibrillation were associated with both stroke recurrence and recurrence-free survival. In the stroke population, 9.1% (95% CI, -2.0 to 20.2) of recurrences were attributable to diabetes and 4.9% (95% CI, -7.3 to 17.2) to atrial fibrillation during the first year after the index stroke. CONCLUSIONS: The cause of stroke recurrence is multifactorial, and the subtypes of index and recurrent strokes are often not identical. Most recurrences remain unexplained by conventional risk factors. PMID- 12750545 TI - Intracellular pathways involved in upregulation of vascular endothelin type B receptors in cerebral arteries of the rat. AB - BACKGROUND AND PURPOSE: Previous studies have shown that contractile endothelin type B (ETB) receptors are upregulated in cerebral arteries after experimental focal cerebral ischemia. The aim of this study was to examine the upregulation of contractile ETB receptors in cerebral arteries after organ culture and to elucidate the intracellular pathways involved. METHODS: Rat middle cerebral arteries (MCAs) were incubated with or without inhibitors. The vessels were mounted in myographs, and the contractile responses to endothelin-1 (ET-1) (ETA and ETB receptor agonist) and sarafotoxin 6c (ETB receptor agonist) were measured. Levels of ETB receptor mRNA were measured with real-time polymerase chain reaction. RESULTS: In fresh MCA, sarafotoxin 6c had no contractile effect. However, after organ culture, a strong concentration-dependent contraction was induced. ET-1 produced a strong contraction, in which the Emax was unaffected by organ culture but the EC50 was decreased with time. The sarafotoxin 6c-induced contraction after 24 hours of organ culture was attenuated by the transcriptional inhibitor actinomycin D and the translational inhibitor cycloheximide as well as the protein kinase C inhibitor Ro-31-8220. Real-time polymerase chain reaction revealed that the mRNA levels of the ETB receptor were increased after organ culture compared with fresh vessels. Actinomycin D and Ro-31-8220 diminished the enhanced mRNA levels considerably. CONCLUSIONS: The results suggest that, in fresh MCA, the ETA receptor is the most prominent subtype, while after organ culture ETB receptors also contribute to the contraction. This upregulation is due to de novo transcription of receptors. Protein kinase C is involved in the upregulation as Ro-31-8220 attenuates the contraction and the mRNA increase. PMID- 12750546 TI - Recommendations for advancing development of acute stroke therapies: Stroke Therapy Academic Industry Roundtable 3. AB - BACKGROUND: The development of acute stroke therapy has proven to be a daunting task, with a few successful and many unsuccessful trials. New strategies need to be considered to enhance the chances for success in future trials. SUMMARY OF REVIEW: The third Stroke Therapy Academic Industry Roundtable (STAIR) conference focused on issues related to increasing the percentage of acute stroke patients who might be included in acute stroke therapy trials and ultimately treated with drugs proven to be effective. A second focus was directed at the need for implementing multimodality stroke trials and potential ways to organize such trials in the near future. Finally, concepts for organizing and implementing acute stroke trials that incorporate current, state of the art trial methodology were discussed. CONCLUSIONS: It is hoped that these suggestions will enhance future stroke trials and the development of effective, new acute stroke treatments that are maximally effective and utilized. PMID- 12750547 TI - Familial intracranial aneurysms: an analysis of 346 multiplex Finnish families. AB - BACKGROUND AND PURPOSE: Genetic risk factors are considered important in the development, growth, and rupture of intracranial aneurysms; however, few have been identified. We analyzed intracranial aneurysm families with at least 2 affected persons and determined relationships between affected persons and assessed the inheritance patterns of aneurysms. METHODS: Families with > or =2 members with verified diagnoses of intracranial aneurysms were recruited from Kuopio and Helsinki, Finland. Families with a diagnosis of other heritable disorders that have associated intracranial aneurysms, such as autosomal dominant polycystic kidney disease, were excluded. RESULTS: We identified 346 Finnish multiplex families with 160 (46.2%) male and 186 (53.8%) female index cases. There were a total of 937 aneurysm cases, with an average of 2.7 cases per family. The majority of the families had only 2 affected relatives (n=206; 59.5%), although there were families with up to 6 (n=10), 7 (n=1), 8 (n=1), or 10 (n=2) affected persons. The affected relatives of the index cases included 108 sisters, 116 brothers, 105 parents, 30 children, 15 grandparents, 102 aunts or uncles, and 64 cousins. Of the 937 affected persons, 569 (60.7%) were alive and available for genetic analysis. Inheritance patterns consistent with autosomal recessiveness were observed in 198 (57.2%), autosomal dominance in 126 (36.4%), and autosomal dominance with incomplete penetrance in 19 (5.5%) of the families. CONCLUSIONS: The collection is the most extensive published to date and extends previous observations of familial aggregation that are consistent with a major gene effect. PMID- 12750548 TI - Editorial comment: Low-dose or moderate-dose anticoagulation: dream or hope for stroke prevention? PMID- 12750550 TI - Telomerase as a target for cancer immunotherapy. AB - Telomerase is the ribonucleoprotein that enables cancer and stem cells to maintain their telomeres, resulting in unlimited proliferative potential. The catalytic component of telomerase in humans, hTERT, is upregulated in nearly 90% of all cancers, making it the most widely expressed marker of malignancy. With the exception of germ cells and stem cells, hTERT is undetectable in somatic human tissues. Together, these properties make telomerase a leading candidate for cancer therapy. Various therapies have been tested in tissue culture and in mouse models utilizing genetic, pharmacological, and immunological approaches. The purpose of this review is to critically examine the role of hTERT in cancer immunotherapy by providing a comparison of the current experiments and a proposal for an innovative method utilizing DNA vaccination. PMID- 12750549 TI - Role of the retinoblastoma protein in differentiation and senescence. AB - The retinoblastoma protein pRb is functionally inactivated in most human cancers. Numerous studies in cell culture and animal models suggest that pRb has a unique ability to encourage and enforce permanent cell cycle withdrawal, consistent with its role as a tumor suppressor protein. This cell cycle withdrawal has a generic component involving repression of transcription of genes required for proliferation. In addition, numerous studies hint at additional specific roles for pRb in differentiation of certain tissue types. Further, pRb appears to play a central role in the process of cellular senescence, a tumorsuppressive process characterized by proliferative arrest and phenotypic changes. Both differentiation and senescence pathways influenced by pRb involve direct and indirect interactions with the core machinery involved in cell-type-specific differentiation and cell shape control. This review focuses on pRb's role as an participant in osteoblast differentiation illustrative of a broader role in terminal differentiation. In addition, novel pathways activated by pRb in its role as an inducer of cellular senescence will be discussed. PMID- 12750551 TI - Why Iressa failed: toward novel use of kinase inhibitors (outlook). AB - A phase III failure of Iressa, an inhibitor of the EGF receptor (EGFR), is viewed as a surprise. With a few exceptions, however, inhibitors of EGFR cannot be effective as a monotherapy in cancers with additional oncogenic changes (downstream of EGFR), which cause mitogen-independent proliferation. In other cases, combining these inhibitors with chemotherapy may lead to antagonism in cancer cells and/or aggravated side-effects. This review discusses why neither levels of EGFR nor doses of Iressa correlate with clinical response. We suggest a novel use of signal transduction inhibitors, including Iressa, to increase therapeutic index by modulating cycle-dependent and apoptosis-inducing chemotherapies. This approach may be most beneficial to patients who do not respond to monotherapy with kinase inhibitors. Development of molecular diagnostics will further diversify these strategies. PMID- 12750552 TI - Mu-calpain activation in beta-lapachone-mediated apoptosis. AB - Beta-lapachone (beta-Lap) triggers apoptosis in a number of human breast and prostate cancer cell lines through a unique apoptotic pathway that is dependent upon NQO1, a two-electron reductase. Recently, our laboratory showed that beta lap-exposed MCF-7 cells exhibited an early increase in intracellular cytosolic Ca(2+) from endoplasmic reticulum stores, and that BAPTA-AM (an intracellular Ca(2+) chelator) blocked these early increases and partially inhibited all aspects of beta-lap-induced apoptosis. We now show that exposure of NQO1 expressing breast cancer cells to beta-lap stimulates a unique proteolytic apoptotic pathway involving mu-calpain activation. No apparent activation of m calpain was noted. Upon activation, mu-calpain translocated to the nucleus concomitant with specific nuclear proteolytic events. Apoptotic responses in beta lap-exposed NQO1-expressing cells were significantly delayed and survival enhanced by exogenous over-expression of calpastatin, a natural inhibitor of mu- and m-calpains. Furthermore, purified mu-calpain cleaved PARP to a unique fragment (approximately 60 kDa), not previously reported for calpains. We provide evidence that beta-lap-induced, mu-calpain-stimulated apoptosis does not involve any known apoptotic caspases; the activated fragments of caspases were not observed after beta-lap exposures, nor were there any changes in the pro-enzyme forms as measured by Western blot analyses. The ability of beta-lap to trigger an apparently novel, p53-independent, calpain-mediated apoptotic cell death further support the development of this drug for improved breast cancer therapy. PMID- 12750553 TI - Calpains can do it alone: implications for cancer therapy. PMID- 12750554 TI - Antitumoral effect of a vaccination procedure with an autologous hemoderivative. AB - Lately, the promising results obtained with autologous cancer vaccines are stimulating new research in the old field of cancer immunotherapy. This paper describes the development of a procedure previously reported by us that is used to obtain an autologous hemoderivative with antitumoral properties. The procedure has been tested in a phase I-II, randomized, controlled clinical trial of 28 cancer patients with different primary malignancies in metastatic and chemotherapy-resistant stages. The histology of the lesions that responded to this treatment was consistent with the characteristic histology observed in malignant lesions treated with a similar antitumoral hemoderivative: proliferation of stromal connective tissue, T-lymphocyte infiltration, and a reduction in the amount of tumor cells and blood vessels. We concluded that vaccination had elicited an immune response because a delayed-type hypersensitivity test made with the autologous hemoderivative produced a significantly more intense response in the responding treated patients. We propose that an immune mechanism acting on tumor cells and/or the regulatory system for stromal growth explains the histological results observed. The use of blood to obtain the immunogen allows vaccination to be repeated, so this method could avoid tumor escape responses due to mutations in the antigen library of the tumor. The results of our study justify further research to optimize the antitumoral effect of vaccination. PMID- 12750555 TI - A new twist on autologous cancer vaccines. PMID- 12750556 TI - The essential similarity of TGFbeta and activin receptor transcriptional responses in cancer cells. AB - The binding of activin and TGFbeta to their respective receptors initiates signals that are carried by common intermediates (Smad proteins) to induce transcriptional activation of downstream genes. Mutations in tumors indicate that both receptor types convey tumorsuppressive signals, among other biologic roles, but their respective sets of transcriptional targets (transcriptomes) and the shared degree of transcriptome similarity are not well explored in these cells. Transcriptome changes were analyzed by gene expression profiling after expression of constitutively active activin type I (ALK4m) and TGFbeta type I (ALK5m) receptors and by variation of Smad4 expression in cancer cells. Eleven of 15 previously reported TGFbeta downstream genes were confirmed to be responsive to TGFb and activin receptors in cancer cells. Expression profiling detected eight of these 11, as well as 13 new Smad4-dependent transcripts. Although Smad4 dependent CDKN1A/p21 induction represents the sole known effector of TGFbeta and activin tumor-suppressor effects, many downstream genes have not yet been evaluated for a suppressive role. A high similarity of TGFbeta and activin responses among the known and new transcriptional target genes indicated an essential redundancy of the two related inputs. This similarity helps relate the mutations seen in both receptor systems and their Smad mediators in human cancers. PMID- 12750557 TI - A tumor-suppressing duo: TGFbeta and activin modulate a similar transcriptome. PMID- 12750558 TI - Dietary glycine inhibits angiogenesis during wound healing and tumor growth. AB - In this study we investigated the effects of glycine on angiogenesis during embryogenesis, wound healing and tumor growth. In chorioallantoic membrane (CAM) assay, glycine (100 microM) inhibited angiogenesis by more than 50%. We studied dietary glycine's effect on fibrin induced wound healing response in a novel (Fibrin Z-chamber) assay. Fibrin within the chamber triggers the healing cascade leading to formation of granulation tissue (GT) rich in blood vessels and stroma. GT was reduced by more than 30% (p < 0.0001) in dietary Glycine groups as compared to control. We found that microvessel density dropped significantly (15%, p < 0.0003) with dietary glycine whereas the other components of GT were unaffected. We evaluated tumor growth delay utilizing Tumor Z-Chamber (fibrin with R3230 mammary adenocarcinoma cells) since tumors take advantage of angiogenesis and matrix formation. We observed that tumor growth decreased by 15% (p < 0.03) and tumor microvessel density dropped by 20% (p < 0.03) with dietary glycine compared to controls. We found that iNOS protein levels were decreased significantly in both GT (24%-57%) and tumor tissue (19-75%). In conclusion, we found that dietary glycine is a potent anti-angiogenic agent that can reduce wound healing and tumor growth through reduction of iNOS expression. PMID- 12750559 TI - Aberration in the expression of the retinoid receptor, RXRalpha, in prostate cancer. AB - There is ample evidence for a role for retinoids in the development and maturation of prostatic epithelium. In recent experiments with conditional disruption of a specific retinoid receptor, namely, RXRalpha in the prostatic epithelium of the mouse, we observed that a major component of retinoid action in the prostate is indeed mediated by RXRalpha. The results clearly indicated that the inactivation of RXRalpha in the prostate epithelium leads to the development of preneoplastic lesions (Huang et al. Cancer Res 62: 4812-9, 2002). To determine the relation of this finding to human prostate cancer, we examined the expression of RXRalpha protein in human prostate cancer cell lines by western blotting and prostate cancer specimens by immunohistochemistry. Relative to the "normal" prostate epithelial cells, there was approximately two- to nine-fold decrease in the full-length 54 kD RXRalpha protein in each of the seven different prostate cancer cell lines tested. Similarly, while RXRalpha immunostaining was uniformly strong in the nuclei of most of the benign prostatic epithelial cells of the thirteen adenocarcinoma specimens tested, a highly heterogeneous pattern of expression was detected in the malignant epithelium, with some areas with low or no staining, others with mostly cytoplasmic staining, and some with both nuclear and cytoplasmic immunoreactivity. To evaluate the effect of RXRalpha modulation on the biologic properties of prostate cancer cell lines, we used a lentivirus expression system to overexpress RXRalpha in CWR22R prostate cancer cells that basally expressed a marginal level of the receptor. The sorted RXRalpha transduced cells were compared to the corresponding vector control cells for proliferative and apoptotic properties. A correlation of reduction of cell growth or increased susceptibility to apoptosis with increases in the level of RXRalpha nuclear receptor was demonstrated. These effects were further enhanced when the cell culture medium was supplemented with a retinoid receptor panagonist, 9-cis retinoic acid. Together, these data support the notion that, like in mouse prostate, loss or reduction of RXRalpha activity might be a critical factor in prostate tumorigenesis in humans. PMID- 12750560 TI - RXRalpha: a novel target for prostate cancer. PMID- 12750561 TI - DF3/MUC1 signaling in multiple myeloma cells is regulated by interleukin-7. AB - The human DF3/MUC1 transmembrane protein is aberrantly expressed in multiple myeloma cells and other B cell malignancies. The regulation of MUC1 in B cells and its potential function as a signaling molecule are unknown. The present results demonstrate that interleukin-7 (IL-7) stimulates MUC1 expression in multiple myeloma cells. The results also demonstrate the IL-7 induces binding of MUC1 to the Lyn tyrosine kinase. The MUC1 C-terminal subunit binds directly to Lyn through interactions with the Lyn SH3 and SH2 domains. Activation of Lyn in response to IL-7 stimulation results in increased tyrosine phosphorylation of the MUC1 C-terminal subunit. In vitro and in vivo studies show that Lyn phosphorylates MUC1, at least in large part, on a YEKV site in the MUC1 cytoplasmic tail. The functional significance of the MUC1-Lyn interaction is supported by the demonstration that Lyn-mediated phosphorylation of MUC1 on YEKV induces binding of MUC1 and the beta-catenin signaling protein. In concert with these results, IL-7 treatment is associated with binding of MUC1 to beta-catenin and targeting of the MUC1-beta-catenin complex to the nucleus. These findings indicate that IL-7 regulates MUC1 expression and function in multiple myeloma cells. PMID- 12750562 TI - Interleukin-7 induces MUC1. PMID- 12750563 TI - Bioluminescent molecular imaging of endogenous and exogenous p53-mediated transcription in vitro and in vivo using an HCT116 human colon carcinoma xenograft model. AB - Real-time p53 activity in tumor cells was detected non-invasively both in vitro and in vivo by bioluminescent imaging. HCT116 colon cancer cells were stably transduced with PG13-luc, a p53 reporter with a Firefly luciferase gene under the control of 13 p53 response elements, together with a Renilla luciferase gene under an MMLV long terminal repeat promoter. Basic conditions for both in vivo and in vitro imaging were explored. Signals from as few as three thousand cells in a 96-well plate were detected following addition of D-luciferin, a substrate of Firefly luciferase at a concentration of 100 microg/ml. Bioluminescence from fifteen thousand cells with PG13-luc inoculated subcutaneously was detected following intravenous injection of D-luciferin at a dose of 100 mg/kg. Intraperitoneal injection serves as an alternative and effective route for D luciferin delivery, although the maximal luminescent intensity was 4-10 times lower than that from intravenous injection. Bioluminescence from Renilla luciferase constitutively expressed in tumor cells was also imaged both in vitro and in vivo and served as an internal control to monitor the physiological state of the cells or tumor volume. Infection of the cells with adenovirus carrying p53 increased the bioluminescent intensity both in vitro and in vivo. Non-invasive imaging of p53 transcriptional activity provides a practical way to monitor the p53 response in cell culture and in animal models. PMID- 12750564 TI - Seeing is believing: visualization of transcriptional activity of p53. PMID- 12750565 TI - Use of lentiviral vectors for delivery of small interfering RNA. AB - Silencing of gene expression by small interfering RNAs (siRNAs) is rapidly becoming a powerful tool for genetic analysis of mammalian cells. Although in the original studies expression of siRNA in mammalian cells was achieved via the transfection of double stranded oligonucleotides, subsequent studies described the use of plasmids to achieve long-term and stable expression of siRNA. Recently, several groups have described the use of retroviral vectors for siRNA delivery. However, retroviral vectors require active cell division for gene transfer and also suffer from the problem of gene-silencing. In this report we have modified a commercially available self-inactivating lentiviral vector for the delivery of siRNA into mammalian cells. We demonstrate the ability of this modified vector to efficiently transfer siRNA into HeLa S3 cells and downregulate p53 expression. Our results suggest that lentiviruses are efficient vectors for delivery of siRNA into mammalian cells. Based on the known ability of these vectors to infect both dividing and non-dividing cells and achieve long-term multilineage gene expression, their use as a therapeutic tool for the delivery of siRNA deserves further study. PMID- 12750566 TI - Stable RNA interference (RNAi) in mammalian cells. PMID- 12750567 TI - Preparing for serendipity. PMID- 12750569 TI - Antibiotic prophylaxis in the surgical patient. AB - Antimicrobial selection for prophylaxis in surgery is based on the site of surgery, likely pathogen involved in addition to the in vitro efficacy, pharmacokinetics and the cost of the drugs used. For example, prophylactic agent in cardiovascular or orthopedic and skin and soft tissue surgery must cover against Staphylococcus aureus as well as the enterobacteriaceae; cephalosporins being adequate. On the other hand, any drug used in cases of urologic surgery must be excreted by the kidney in an active state and should also be active against E. coli, a common uropathogen such as any cephalosporins or penicillin. Drugs like macrolides and tetracyclines accumulate in the prostate and are good for prostate surgery by they are not excreted well by the kidney and therefore useless for urological prophylaxis. It is important to note that even treating minor infections in a neurosurgical patient, we must use an antibiotic that cross blood-brain barrier otherwise meningitis might develop. In cases of gastrointestinal tract, surgery on the colon and appendix requires special coverage against anaerobic bacteria especially Bacteroides fragilis. On the other hand surgery on the stomach, gall-bladder and upper two thirds of small intestine, it is adequate to use drugs to cover aerobic bacteria such as E. coli. Most studies show that a single most effective antibiotic is enough and it is unnecessary to use two or more drug combinations. PMID- 12750570 TI - [Quantitative Detection of cytomegalovirus (CMV) DNA in leukocytes of patients with HIV infection by using the Digene Hybrid Capture System (DHCS)] AB - The aim of the present study was to evaluate the usefulness of the Digene Hybrid Capture System (DHCS) for the detection and quantitation of cytomegalovirus (CMV) DNA in 95 blood samples from 57 HIV-positive patients with low CD4+ T-cell count (<100 cells/ l). The DHCS was compared with pp65 antigenemia assay and the results were correlated with active CMV disease, anti-CMV therapy and occurrence of disease relapses. Our data suggest that the detection of CMV DNA by DHCS seems to be a rapid, sensitive and specific assay for the diagnosis of CMV disease in HIV-infected patients, showing a good correlation with pp65 antigenemia assay. Overall, the DHCS provides a quantitative and objective measure of CMV activity in leukocytes and it may also represent a useful tool for the monitoring of anti CMV therapy. PMID- 12750571 TI - [Evaluation of in-patient stay appropriateness and of functional independence in a sample of HIV-infected subjects] AB - The evaluation of in-patient stay appropriateness and of functional independence in subjects infected with HIV has been carried out in a sample of patients admitted to an infectious diseases hospital. The evaluation of in-patient stay appropriateness has been performed by means of the Appropriateness Evaluation Protocol, whereas functional levels have been assessed by administering the Functional Independence Measure. In 25% of patients, the appropriateness criteria for in-patient stay were compatible with an extra-mural approach. The availability of day-hospital and of home care assistance is a crucial factor to avoid needless and expensive in-patient stay. PMID- 12750572 TI - Comparison of amoxicillin with second and third generation cephalosporins in the treatment of acute otitis media. AB - The aim of the present study was to compare the efficacy and the tolerability of three different antibiotic regimens for the treatment of acute otitis media in paediatric patients. 75 children, age range 6 months-6 years, divided into three groups, were involved in the study, each group consisting of 25 randomly assigned children (Group 1, amoxicillin 40 mg/kg tid per os for 10 days; Group 2, cefuroxime axetil 30 mg/kg bid per os for 10 days; ceftriaxone 50 mg/kg single dose i.m.). No statistically significant difference was noted in the clinical efficacy among the different groups. Although amoxicillin is the drug of choice in paediatric otitis media, single dose ceftriaxone might be considered as an interesting alternative regimen when ease of administration and cost of therapy are considered. PMID- 12750573 TI - [Acute reversible degenerative myopathy complicating Lyme disease: description of a case] AB - Lyme's disease is a commonly found disorder whose etiological agent is a spirochete named Borrelia burgdorferi. Its clinical presentation is usually well known, and three stages are described. However it may present symptoms which may resemble many infectious or autoimmune diseases as well. The aim of the present study is to describe a case of first stage Lyme's disease whose main clinical picture is muscoloskeletal involvement, which may mimic dermatomyositis. Many subtle characteristics (age of onset, atypical cutaneous rash) together with the involvement of muscular masses other than scapular or pelvic ones, and the bio humoral positivity for anti-Borrelia antibodies allowed correct diagnosis to be made. We emphasize that instrumental and invasive examinations, such as electromyography and muscular biopsy, were not diagnostic in such cases. PMID- 12750574 TI - [Cerebral hemorrhagic infarction caused by tubercular arteritis in HIV positive patient: a case report] AB - A case of cerebral hemorrhagic infarction caused by tubercular arteritis complicating the course of a tubercular meningitis in an HIV-positive patient is described. The atypical clinical course and histopathologic findings of this rare deadly complication of specific meningeal infection are underlined. PMID- 12750575 TI - [Disulfiram-induced hepatitis] AB - Overt disulfiram-induced hepatitis is rare; it has a high mortality rate, especially when the etiology is non suspected and when the treatment is not promptly discontinued. The Authors describe a case of disulfiram-induced acute hepatitis and emphasize close clinical and biochemical monitoring. PMID- 12750576 TI - [The struggle against the variolous poison (Part 2)] AB - Not available PMID- 12750577 TI - [Sexually transmitted diseases in the History of Medicine] AB - Classic sexually transmitted diseases (syphilis, gonorrhea, condyloma acuminatum, venereal ulcera) were known since ancient times. Being considered as a "divine punishment" for impure intercourses, they were generally mistaken one for another (except for condylomatosis) until the different pathogenic agents were identified in the course of 19th Century. It was only with the introduction of antibiotics that their incidence drastically fell, at least up to this last decade, as a number of factors determined their fresh outbreak. PMID- 12750578 TI - Disordered eating, menstrual irregularity, and bone mineral density in female runners. AB - PURPOSE: To examine the relationships between disordered eating, menstrual irregularity, and low bone mineral density (BMD) in young female runners. METHODS: Subjects were 91 competitive female distance runners aged 18-26 yr. Disordered eating was measured by the Eating Disorder Inventory (EDI). Menstrual irregularity was defined as oligo/amenorrhea (0-9 menses per year). BMD was measured by dual x-ray absorptiometry. RESULTS: An elevated score on the EDI (highest quartile) was associated with oligo/amenorrhea, after adjusting for percent body fat, age, miles run per week, age at menarche, and dietary fat, (OR [95% CI]: 4.6 [1.1-18.6]). Oligo/amenorrheic runners had lower BMD than eumenorrheic runners at the spine (-5%), hip (-6%), and whole body (-3%), even after accounting for weight, percent body fat, EDI score, and age at menarche. Eumenorrheic runners with elevated EDI scores had lower BMD than eumenorrheic runners with normal EDI scores at the spine (-11%), with trends at the hip (-5%), and whole body (-5%), after adjusting for differences in weight and percent body fat. Runners with both an elevated EDI score and oligo/amenorrhea had no further reduction in BMD than runners with only one of these risk factors. CONCLUSION: In young competitive female distance runners, (i) disordered eating is strongly related to menstrual irregularity, (ii) menstrual irregularity is associated with low BMD, and (iii) disordered eating is associated with low BMD in the absence of menstrual irregularity. PMID- 12750579 TI - Bone mass of asian adolescents in China: influence of physical activity and smoking. AB - INTRODUCTION/PURPOSE: Research addressing the role of biology and behavior on bone development during times of peak bone acquisition in adolescence is limited. The present investigation was conducted to address the influence of body composition (lean body mass, fat mass), menarche, leisure physical activity, sports team participation, smoking, and second-hand smoke on skeletal mass of a unique sample of Asian adolescents in China. METHODS: A total of 166 girls and 300 boys (ages 12-16 yr) participated in this study. Bone mineral density (BMD) and content (bone mineral content (BMC)) of the forearm and the os calcis were measured using dual energy x-ray absorptiometry (DXA); lean body mass (LBM) and fat mass were estimated by bioelectrical impedance analysis (BIA); grip strength was measured by isometric dynamometry. Menarche, leisure physical activity, sports team participation, and active and passive smoking were determined using questionnaire. RESULTS: In girls, a total of 44% of the variance in forearm BMC was attributed to a model which included LBM (32%), time since menarche (10%), and age (2%); heel BMC was best predicted by LBM alone (42%), with no significant contribution by other variables. In boys, a total of 39% of the variance in forearm BMC was attributed to a model which included LBM (28%), age (5%), sports team participation (4%), height (1%), and fat mass (1%); heel BMC was best predicted by LBM (50%) and height (3%), accounting for 53% of the variance. CONCLUSION: The findings of this study suggest that lean body mass is the primary determinant of bone mass in Chinese adolescents. Menarche is also an important contributor in girls, whereas age and sports team participation are secondary predictors of bone mass in boys. PMID- 12750580 TI - Esophageal reflux in conditioned runners, cyclists, and weightlifters. AB - INTRODUCTION: Gastroesophageal reflux disease is a disorder in which gastric contents move from stomach to esophagus. Exercise is a recognized contributing factor to reflux in healthy volunteers and is reported to be proportional to exercise intensity and the type of exercise. Our aim was to explore changes in physiology occurring in conditioned runners, cyclists, and weightlifters. METHODS: Ten subjects from each sport with >3-month history of exercise-induced heartburn were enrolled. Subjects underwent evaluation of fasting and fed esophageal pH, heart rate, GI symptom, and perceived exertion during standardized exercise routines at 65% (60 min) and 85% (20 min) of their maximal capabilities. RESULTS: Weightlifters experienced the most heartburn and reflux: 18.51 +/- 17.34% time esophageal pH Unfit" (the referent group), the estimated relative risks from the simulations for men who were reclassified as fit (i.e., "Unfit-->Fit") were 0.57 for total mortality and 0.52 for CVD mortality, and for men who remained classified as fit ("Fit-->Fit"), they were 0.33 for total mortality and 0.20 for CVD mortality. CONCLUSION: The imprecision of the fitness measurement alone (i.e., measurement error) is sufficient to produce the reported ACLS risk reductions in initially unfit men who get reclassified as fit in a subsequent clinic visit. This statistical artifact will apply to other studies that use this design. PMID- 12750582 TI - Changes in serum biochemical responses during cardiac rehabilitation. AB - PURPOSE: The objective of the study was to establish the difference in the exercise response profile of the enzymes lactate dehydrogenase (LDH), lactate dehydrogenase isoenzyme 1 (LDH-1), creatine kinase (CK), and creatine kinase polypeptide subunit MB (CK-MB) after a 12-wk cardiac rehabilitation exercise program. METHODS: Nineteen male nonblockade recent postmyocardial infarction patients (PMIP), aged 60.7 +/- 6.5 yr, performed a graded exercise test on a motorized treadmill until volitional cessation or reaching any of the American College of Sports Medicine criteria. After this, a 12-wk exercise-based cardiac rehabilitation program was adopted at a frequency of 3x wk-1. Each subject was then retested using the same protocol and intensity. Blood samples were collected by venipuncture from the antecubital vein into Vacutainer tubes preexercise, immediately, postexercise, and 24 h later. The blood samples were then centrifuged to obtain serum before being transported to the laboratory for enzymatic analysis by conventional methods. RESULTS: a) a 12-wk cardiac rehabilitation exercise program had no influence on the serum activities of enzymes at rest; b) the same exercise reduced the enzyme activity level after training; and c) LDH and CK values returned to normal faster after the program. CONCLUSIONS: These findings provide further evidence for the benefits of exercise based cardiac rehabilitation and demonstrate that an exercise test provokes a temporary serum enzyme response, which does not reflect myocardial damage. PMID- 12750583 TI - Noninvasive monitoring of cardiac output during exercise by inductance cardiography. AB - INTRODUCTION/PURPOSE: Inductance cardiography is a noninvasive technique that monitors changes in cardiac output from an inductance plethysmographic transducer encircling the chest at the level of the heart. The method has been previously validated in supine patients at rest by comparisons to thermodilution. Our purpose was to investigate whether the technique can be employed during bicycle exercise. METHODS: We simultaneously measured cardiac output by inductance cardiography and by two gas exchange methods based on the Fick principle during upright cycle ergometry in healthy volunteers. RESULTS: In 11 subjects, comparisons of changes in cardiac output by inductance cardiography agreed well with values measured by carbon dioxide rebreathing during a steady-state exercise protocol. In 12 subjects, cardiac output changes measured by inductance cardiography and an oxygen uptake method agreed closely during a progressive ramp exercise protocol to exhaustion. The bias (mean difference to reference methods) and limits of agreement (+/-2 SD of bias) for estimation of relative changes in cardiac output by inductance cardiography were 1% +/- 21% in 67 comparisons to the carbon dioxide rebreathing technique, and 0% +/- 22% in 98 comparisons to the oxygen uptake method. CONCLUSION: In healthy subjects, inductance cardiography accurately and unobtrusively estimates changes in cardiac output during bicycle exercise in comparison to gas exchange methods. PMID- 12750584 TI - Short-term immobilization after eccentric exercise. Part I: contractile properties. AB - PURPOSE: The purpose of this study was to examine the compound muscle action potential (M-wave) and evoked contractile properties of immobilized muscle after high-force eccentric exercise. We believed that changes in these variables would contribute to the enhanced recovery of maximal voluntary force observed after short-term immobilization of damaged muscle. We hypothesized that immobilization after eccentric exercise would result in an enhanced M-wave and a change in contractile properties toward characteristics of faster muscle fibers. METHODS: Twenty-five college-age males were matched according to force loss after 50 maximal eccentric contractions of the elbow flexors and placed into an immobilization (IMM, N = 12) or control (CON, N = 13) group. IMM had their arm immobilized at 90 degrees and secured in a sling during a 4-d treatment. Maximal isometric torque (MVC) was assessed at baseline and for 8 d after treatment. M wave and evoked contractile properties of the muscle (twitch torque [TT], maximal rate of torque development [MRTD], time to peak torque [TPT], and one-half relaxation time [HRT]) were assessed at baseline and for the first 5 d after treatment. RESULTS: Immediately postexercise, MVC was reduced 43% and 42% in IMM and CON, respectively. Recovery of MVC was significantly greater in IMM during recovery (P < 0.05), 95% of baseline MVC compared with 83% in CON. M-wave was reduced 32%, and all contractile properties were altered immediately postexercise. M-wave, MRTD, TPT, and HRT were not significantly different between groups during recovery (P > 0.05). TT demonstrated enhanced recovery in IMM (P < 0.05). CONCLUSIONS: Short-term immobilization after eccentric exercise resulted in enhanced recovery of maximal voluntary force. However, enhanced force recovery cannot be explained by muscle activation and evoked contractile properties of the muscle. PMID- 12750585 TI - Short-term immobilization after eccentric exercise. Part II: creatine kinase and myoglobin. AB - PURPOSE: Although creatine kinase (CK) is commonly used as a marker of muscle damage, there is large variability in the response to exercise. We previously found short-term immobilization blunted the rise in plasma CK after eccentric exercise, suggesting subsequent movement of damaged muscle may contribute to variability. We hypothesized that immobilization decreases lymphatic transport of CK from damaged muscle, blunting the CK response. In this study, we compared changes in plasma CK and myoglobin (Mb), as Mb is released from damaged muscle directly into the bloodstream whereas CK is released first into the lymph. METHODS: Twenty-five college-age males were matched according to force loss after 50 maximal eccentric contractions of the elbow flexors and placed into an immobilization (IMM, N = 12) or control (CON, N = 13) group. IMM had their arm immobilized at 90 degrees and secured in a sling for 4 d (treatment). Venipuncture was performed during baseline, treatment, and for 5 d after treatment (recovery) to assess plasma CK activity and Mb. Urine specific gravity (USG) and muscle activity (ACT) were assessed. RESULTS: Immobilization significantly blunted increases in CK activity (IMM: 955 +/- 316 IU.L-1 vs CON: 2884 +/- 1083 IU.L-1; P < 0.05) but not increases in Mb (IMM: 712 +/- 278 ng.mL-1 vs CON: 891 +/- 253 ng.mL-1; P > 0.05). There were no differences in USG between groups over time (P > 0.05) and no group differences in ACT (P > 0.05). CONCLUSIONS: Short-term immobilization after eccentric exercise blunted the CK response but not the Mb response, suggesting lymphatic transport of CK may be responsible. Because hydration status and muscular activity after exercise were not different between groups, the blunted CK response was likely due to inactivation of CK activity before entering circulation. PMID- 12750586 TI - The effects of ergogenic compounds on myogenic satellite cells. AB - PURPOSE: A series of studies were conducted in which compounds commonly shown to be ergogenic aids for strength athletes if taken orally were evaluated for their ability to directly induce postnatal muscle stem cell proliferation or differentiation/fusion in vitro. METHODS: Compounds tested were creatine monohydrate, creatine pyruvate, L-glutamine, dehydroepiandrosterone (DHEA), androstenedione, Ma Huang (Ephedra sinensis) extract, and Zhi Shi (Citrus aurantium) extract. Dulbecco's modified eagle medium, supplemented with minimal levels of serum and antibiotics, was used as the initial vehicle for the test compounds. Subsequently, a defined treatment medium termed ITTC was used. Satellite cells were exposed to the test compounds for the indicated times and then evaluated by counting mononucleated and multinucleated (fused) cells. RESULTS: In serum-containing media, none of the treatment groups displayed increased proliferation over that of the control. However, in the differentiation cultures, 0.10% creatine monohydrate increased differentiation over that of the control cultures. When 0.10% creatine monohydrate was added to defined media formulations, all treatments but one demonstrated increased differentiation over the 0.5% serum control. Time course experiments, which followed the effect of 0.10% creatine monohydrate contained in ITTC defined media over 120 h, suggested that cells exposed to this treatment differentiated earlier and to a greater level than cells exposed to ITTC alone. CONCLUSIONS: Creatine in the monohydrate form induced differentiation of myogenic satellite cells. Other agents examined did not increase satellite cell proliferation or differentiation. These results provide initial evidence for a mechanistic understanding of observed effects in vivo of increased muscular size and strength from creatine supplementation. PMID- 12750587 TI - Skeletal muscle glucose uptake response to exercise in trained and untrained men. AB - PURPOSE: Endurance training enhances skeletal muscle glucose uptake at rest, but the responses to different exercise intensities are unknown. In the present study, we tested whether glucose uptake is enhanced in trained men during low-, moderate-, and high-intensity exercise as compared with untrained men. METHODS: Seven trained and untrained men were studied without any dietary manipulation during bicycle exercise at relative intensities of 30%, 55%, and 75% of maximal oxygen consumption ([OV0312]O(2max)) on three separate days. Glucose uptake in the quadriceps femoris muscle was directly measured using positron emission tomography (PET) and 18F-fluoro-deoxy-glucose ([18F]FDG). [18F]FDG was injected 10 min after the start of the exercise. Thereafter exercise was continued for another 25 min. PET scanning was conducted immediately after completion of the exercise. The measured glucose uptake values reflect the situation during exercise due to chemical characteristics of the [18F]FDG. RESULTS: Muscle glucose uptake increased from 30% to 55% [OV0312]O(2max) intensity exercise similarly in both groups (P < 0.05). However, from 55% to 75% [OV0312]O(2max) intensity exercise, only athletes were able to further enhance glucose uptake. Furthermore, at highest intensity, glucose uptake was significantly higher in trained than in untrained men (236.6 +/- 29.6 vs 176.3 +/- 22.4 micromol.kg-1.min-1, P < 0.05). There were no differences in plasma glucose, insulin, or lactate in any time point at 75% [OV0312]O(2max) intensity between groups. CONCLUSIONS: These results show that skeletal muscle glucose uptake is higher in trained than in untrained men at high relative exercise intensity, although at lower relative exercise intensities no differences are observed. Thus, endurance training improves the capacity of contraction-induced glucose uptake in skeletal muscle. PMID- 12750588 TI - Free amino acid pool and muscle protein balance after resistance exercise. AB - PURPOSE: The aim of this study was to assess the effects of a resistance exercise session (RES) on free amino acid concentrations and protein synthesis and breakdown of the vastus lateralis (VL) muscle during recovery in male subjects. METHODS: Both the exercise group (EG) and the control group (CG) consisted of six healthy physically active men. On the experiment day in fasting conditions, a stable isotopic tracer of L-[ring-2H(5)] phenylalanine was infused and EG started a heavy 50-min hypertrophic RES for lower extremities after 55 min of infusion. At the same time, CG was at rest. During recovery of 195 min after RES, several blood samples were drawn from the femoral artery (FA) and the femoral vein (FV) and muscle samples from the VL muscle. The enrichment was analyzed by GC/MS and leg muscle amino acid kinetics determined by three-pool compartment model between FA, FV, and VL. RESULTS: During recovery at 60 min after RES, there was no difference in muscle protein synthesis or muscle protein breakdown between the groups, but at 195 min, both muscle protein synthesis (P < 0.05) and muscle protein breakdown (P < 0.05) were increased in EG compared with CG. The protein net balance was negative and similar in both groups. Simultaneously in serum concentrations, there was a decrease in leucine (P < 0.05) associated with an increase in aspartate (P < 0.05). Furthermore, the exercise-induced increase in alanine concentration decreased both in serum and muscle. CONCLUSION: In fasting conditions, protein net balance is negative and RES induces an increase in muscle protein synthesis and breakdown at 195 min but not yet at 60 min of recovery. PMID- 12750589 TI - A longitudinal study on smoking in relationship to fitness and heart rate response. AB - PURPOSE: Smoking has been shown to be associated with impaired cardiovascular fitness and reduced heart rate response to exercise. It is not known whether these associations are present in adolescence and young adults, and whether they change over time. METHODS: Maximal oxygen uptake ([OV0312]O(2max)), maximum treadmill slope (Slope(max)), resting heart rate (HR(rest)), heart rate at submaximal exercise (HR(submax)), heart rate reserve (HRR), and maximum heart rate (HR(max)) were measured one to nine times between ages 13 and 36 in 298 male and 334 female participants of the Amsterdam Growth and Health Longitudinal Study. Generalized estimating equation analyses were used to study the longitudinal relationship between smoking and cardiovascular fitness and heart rate response to exercise, whereas linear regression analyses were used to study the reversibility of smoking effects at age 36. RESULTS: Moderate to heavy smoking (>/=10 g of tobacco per day) was longitudinally and negatively related to [OV0312]O(2max), Slope(max), HR(submax), and HR(max). With increasing age, the negative relationship between smoking and [OV0312]O(2max), Slope(max), and HR(max) became stronger in males. Cross-sectional analyses suggested that the adverse effects of smoking were reversible in 36-yr-old males. CONCLUSION: Cardiovascular fitness and heart rate response to exercise are already reduced in young healthy smokers. In men, the adverse effects of smoking become stronger with increasing age but appear to be reversible at age 36. PMID- 12750590 TI - Validation of the telephone and in-person interview versions of the 7-day PAR. AB - PURPOSE: To validate the 7-d Physical Activity Recall (PAR) telephone interview version and its activity intensity categories. METHODS: Seventy-four adults (47 women, 27 men), ranging in age (18-67) and activity levels, were interviewed by phone and in-person using the same PAR protocol. Each participant wore a TriTrac R3D accelerometer for 10 d. Validity was assessed by comparing the phone and in person PAR interviews with the TriTrac-R3D data. RESULTS: Sixty-nine adults (44 women, 25 men) were used for all statistical analyses. Intraclass correlations between the two PAR interviews for total minutes per week of activity were r = 0.96, and r = 0.94 for moderate, r = 0.97 for hard, and r = 0.97 for very hard intensity activities. Pearson product moment correlations between the phone PAR and TriTrac-R3D for total minutes per week of physical activity were r = 0.43, and r = 0.31 for moderate, r = 0.39 for hard, and r = 0.78 for very hard intensity activities. Pearson correlations between the in-person PAR and TriTrac R3D for total minutes per week of physical activity were r = 0.41, and r = 0.33 for moderate, r = 0.43 for hard, and r = 0.74 for very hard intensity activities. Participants overestimated the amount of physical activity in both interviews as compared with the TriTrac-R3D. CONCLUSION: The phone and in-person versions of the PAR are equivalent measures for self-reported physical activity. Regardless of age, body mass index, or physical activity level both interview methods had similar estimates for total minutes per week of moderate, hard, and very hard activity. Correlations between each interview method and the TriTrac-R3D were lower for moderate and hard activities as compared with very hard activities. PMID- 12750591 TI - Longitudinal analysis of lactate threshold in male and female master athletes. AB - PURPOSE: The lack of relationship between lactate threshold (LT) and running performance in older runners, and the increase in LT with age, has not been previously studied in a longitudinal design. We evaluated the cross-sectional and longitudinal changes in LT with age and compared the changes in LT with changes in performance variables. METHODS: Fifty-one male and 23 female runners (39-77 yr) performed two graded treadmill exercise tests with minute-by-minute venous blood lactate analysis, separated by 5.8 +/- 1.6 yr (mean +/- SD). Body composition was determined by hydrodensitometry and training history by questionnaire. RESULTS: There was no change in LBM over time, but significant decreases in [OV0312]O(2max) and training volume irrespective of age and gender (P < 0.05). LT as a percent of [OV0312]O(2max) increased with age (P < 0.05), demonstrated poor stability over time (r = 0.29, P = 0.01) compared with other parameters measured, and changes in LT were not related to changes in fitness or performance. CONCLUSION: Based upon these findings, we conclude that the LT may be less precise than [OV0312]O(2max) or performance in the prescription of exercise intensities or as an evaluation tool in older individuals. PMID- 12750592 TI - Energy cost of free technique and classical cross-country skiing at racing speeds. AB - PURPOSE: First, to measure the O(2) uptake ([OV0312]O(2)) and the blood lactate concentration during cross-country skiing at competition speed. Second, to compare these entities for the free technique and the classical technique. Further, to establish the subjects' [OV0312]O(2max) and the [OV0312]O(2) corresponding to the onset of blood lactate accumulation (OBLA, 4 mmol.L-1) during skiing, and finally to relate these entities to those of treadmill running. METHODS: Five high-level female junior cross-country skiers with a mean [OV0312]O(2max) of 63 mL.kg-1.min-1 served as subjects in five tests: examination of the [OV0312]O(2max) and the [OV0312]O(2) corresponding to the OBLA during up hill cross-country skiing (both styles), inclined treadmill running, and a 6-km simulated cross-country ski race (both styles). RESULTS: The [OV0312]O(2max) obtained during up-hill cross-country skiing did not differ from that during treadmill running, nor did it differ between the two skiing styles. The peak heart rate was significantly lower during uphill cross-country skiing than during treadmill running. During the simulated competitions, the [OV0312]O(2) averaged 84% of the [OV0312]O(2max) or 95% of the [OV0312]O(2) at the OBLA found for uphill skiing. CONCLUSION: High-level female junior cross-country skiers are unable to ski at intensities close to their [OV0312]O(2max) or maintain an average intensity above that corresponding to their OBLA even during races lasting less than 25 min. Thus, training at intensities around the OBLA may be particularly relevant for cross-country skiers. PMID- 12750593 TI - Pattern of energy expenditure during simulated competition. AB - PURPOSE: To determine how athletes spontaneously use their energetic reserves when the only instruction was to finish in minimal time, and whether experience from repeated performance changes the strategy of recreational athletes. METHODS: Recreational road cyclists/speed skaters (N = 9) completed three laboratory time trials of 1500 m on a windload braked cycle. The pattern of energy use was calculated from total work and from the work attributable to aerobic metabolism, which allowed computation of anaerobic energy use. Regional level speed skaters (N = 8) also performed a single 1500-m time trial with the same protocol and measurements. RESULTS: The serial trials were completed in (mean +/- SD) 133.8 +/ 6.6, 133.9 +/- 5.8, 133.8 +/- 5.5 s (P > 0.05 among trials); and in 125.7 +/- 10.9 s in the skaters (P < 0.05 vs cyclists). The [OV0312]O(2peak) during the terminal 200 m was similar within trials (3.23 +/- 0.44, 3.34 +/- 0.44, 3.30 +/- 0.51 (P > 0.05)) versus 3.91 +/- 0.68 L.min-1 in the skaters (P < 0.05 vs cyclists). In all events, the initial power output and anaerobic energy use was high and decayed to a more or less constant value ( approximately 25% of peak) over the remainder of the event. Contrary to predictions based on an assumed "all out" starting strategy, the subjects reserved some of their ability to perform anaerobic work for a terminal acceleration. The total work accomplished was not different between trials (43.53, 43.78, and 47.48 kJ in the recreational athletes, or between the cyclists and skaters (47.79 kJ). The work attributable to anaerobic sources was not different between the rides (20.67, 20.53, and 21.12 kJ in the recreational athletes). In the skaters, the work attributable to anaerobic sources was significantly larger versus the cyclists (24.67 kJ). CONCLUSION: Energy expenditure during high-intensity cycling seems: 1) to be expended in a manner that allows the athlete to preserve an anaerobic energetic contribution throughout an event, 2) does not appear to have a large learning effect in already well trained cyclists, and 3) anaerobic energy expenditure may be the performance discriminating factor among groups of athletes. PMID- 12750594 TI - Thermal insulation and evaporative resistance of football uniforms. AB - PURPOSE: To measure the thermal resistance (R(t)) and evaporative resistance (R(e,t)) of five different configurations of football uniforms commonly worn for American football practices and games. METHODS: The R(t) (insulation) of the ensembles was measured using an electrically heated manikin in thermal equilibrium with its surroundings. For the R(e,t) determinations, the manikin was covered with a cotton knit "skin" and sprayed with distilled water to simulate sweat-saturated skin. RESULTS: Compared with reference values for a T-shirt and shorts ensemble (R(t) = 0.140 m2. degrees C.W-1, R(e,t) = 0.020 m2.kPa-1.W-1), the R(t) of football uniforms ranged from 0.178 m2. degrees C.W-1 (1.15 clo) for a practice configuration of shorts, shoulder pads, practice jersey, and helmet to 0.233 m2. degrees C.W-1 (1.50 clo) for a full cold-weather uniform. Associated R(e,t) values ranged from 0.027 to 0.039 m2.kPa-1.W-1. CONCLUSION: Football uniforms contribute significantly to the heat load on a player. The thermal and evaporative resistance data presented in this paper can be used in the solution of heat balance equations to predict physiological responses of football players. PMID- 12750595 TI - Modeling the training-performance relationship using a mixed model in elite swimmers. AB - PURPOSE: The aim of this study was to model the relationship between training and performance in 13 competitive swimmers, over three seasons, and to identify individual and group responses to training. METHODS: A linear mixed model was used as an alternative to the Banister model. Training effect on performance was studied over three training periods: short-term, the average of training load accomplished during the 2 wk preceding each performance of the studied period; mid-term, the average of training load accomplished during weeks 3, 4, and 5 before each performance; and long-term, weeks 6, 7, and 8. RESULTS: Cluster analysis identified four groups of subjects according to their reactions to training. The first group corresponded to the subjects who responded well to the long-term training period, the second group to the long- and mid-term periods, the third to the short- and mid-term periods, and the fourth to the combined periods. In the model, the intersubject differences and the evolution over the three seasons were statistically significant for the identified groups of swimmers. Influence of short-term training was negative on performance in the four groups, whereas mid- and long-term training had, on the average, a positive effect in three groups out of four. Between seasons 1 and 3, the effect of mid term training declined, whereas the effect of long-term training increased. The fit between real and modeled performances was significant for all swimmers (0.15 RCP). Total volume and intensity were integrated as a single variable. The score for volume x intensity in each phase was computed by multiplying the accumulated duration in this phase by a multiplier for this particular phase. The total score for Tour and Vuelta was obtained by summating the results of the three phases. RESULTS: The total loads (volume x intensity) did not significantly differ between the two races (P > 0.05), despite a significantly longer total exercise time of the Tour (P < 0.05) (5552 +/- 176 vs 5086 +/- 290 min). CONCLUSION: The physiological loads imposed on cyclists' bodies do not differ between the Tour and Vuelta, despite the longer duration of daily stages in the former race. PMID- 12750602 TI - Need to evaluate regression to the mean. PMID- 12750601 TI - A methodology to assess the accuracy of a portable metabolic system (VmaxST). AB - PURPOSE: Validity of a portable metabolic system (VmaxST) was investigated during gas exchanges simulations by a mechanical system (GESS) and during human exercise. METHODS: Three tests were conducted while gas exchanges were measured continuously by VmaxST. Test 1 was composed of six simulations of gas exchanges during steady-state exercise (20 min at VE = 80 L.min-1). Test 2 was composed of seven simulations of gas exchanges during incremental exercise (VO(2) from 300 to 5600 mL.min-1). In the human trial, 11 subjects performed an incremental running exercise on a treadmill while gas exchanges were measured at the end of each stage with the Douglas bag method (DBM). RESULTS: Test 1 showed that the VmaxST measurements were stable, despite inaccurate measurements of gas concentrations at the start of the test. During test 2, the mean error (difference between measured and predicted value) and the upper and lower limits of agreement were 8.0%, -12.6%, and -3.4% for VO(2); -4.6%, -12.0%, and +2.8% for VCO(2); and 0.7%, -4.7%, and +3.3% for VE. During the human trial, no significant difference was shown between VO(2) measured by VmaxST and by DBM at any stage of exercise. The mean difference and the upper and lower limits of agreement between the VmaxST and the DBM measurements were -0.5%, -14.3%, and +13.3% for VO(2); -6.3%, 20.9%, and +8.3% for VCO(2); and -9.9%, -25.5%, and +5.7% for VE. CONCLUSIONS: The use of GESS showed that measurements of VO(2) by VmaxST could be biased in a standardized condition. In more realistic condition of use, this bias was lower but the accuracy of measurements was impaired. PMID- 12750604 TI - Occupational asthma: a valid model for adult asthma? AB - PURPOSE OF REVIEW: Developments in the understanding of causes and natural history of occupational asthma may allow improved primary, secondary and tertiary preventive strategies for occupational asthma. This may also lead to improved understanding of preventable contributing factors to the development and severity of nonoccupational asthma. RECENT FINDINGS: Animal studies have demonstrated the opportunity to identify chemical sensitizers relevant to asthma. Studies of genetic markers in occupational asthma pose logistic difficulties, but preliminary studies suggest that glutathione S-transferase genotypes may predispose to development of occupational asthma induced by diisocyanates and these have also been implicated in nonoccupational asthma. Some occupational sensitizers/irritants are also found outside the workplace and may be relevant in nonoccupational asthma, for example cleaning agents, epoxy glues, hairdressing products. Accidental exposures to high concentrations of respiratory irritants have the potential to induce new asthma as well as aggravate underlying asthma in both occupational and nonoccupational settings. SUMMARY: Better understanding of the pathogenesis of occupational asthma is important for affected workers, and also has potential relevance for nonoccupational asthma. PMID- 12750605 TI - Recent developments in airway and nose occupational sensitizers. AB - PURPOSE OF REVIEW: To describe the latest developments in the field of occupational asthma and occupational rhinitis in 2001 and 2002. RECENT FINDINGS: Several surveillance programs of occupational diseases, such as Observatoire National des Asthmes Professionnels in France, Surveillance of work-related and Occupational Respiratory Diseases in South Africa (SORDSA), Surveillance of Work related and Occupational Respiratory Diseases (SWORD) in UK, have reported on the frequency of occupational asthma. The causative agents were mainly flour, isocyanates and latex. The common methods of diagnosis - questionnaires, cutaneous tests, Peak Expiratory Flow Rate (PEFR), bronchial hyperresponsiveness still create controversy. In addition, the specific bronchial challenge, the classical gold standard of diagnosis, has its limitations since it cannot be performed in every case. Other methods have been assessed as inflammatory markers in induced sputum. Occupational rhinitis appears to be a poorly diagnosed condition. SUMMARY: Further studies are expected to explore the effect of environmental control and medical surveillance. The key to successful management of occupational asthma and occupational rhinitis may be prospective surveillance of the occurrence of specific IgE antibodies before the onset of allergic symptoms. PMID- 12750606 TI - Use of the specific challenge in the diagnosis of occupational asthma: a 'gold standard' test or a test not used in current practice of occupational asthma? AB - PURPOSE OF REVIEW: This review, an assessment of the role of inhalational challenge testing in the diagnosis of occupational asthma, focuses on the difficulties associated with making the correct diagnosis of occupational asthma. This report contrasts the apparent discrepancy between the clinical diagnosis and the diagnosis made by inhalational challenge testing, the 'gold standard'. This disparity has been pointed out by a number of authors, yet this approach to making the diagnosis of this illness continues. RECENT FINDINGS: Because of the disparity between the clinical mode for diagnosis and using specific challenge testing for diagnosis, awarding compensation to a worker based on the clinical diagnosis of occupational asthma, or using this clinical approach to identify the incidence or prevalence of occupational asthma in a population, is suspect. In the absence of specific inhalational challenge, physicians have attempted to understand changes in flow rates over time through the use of serial peak-flow assessments, a relatively cost-effective way to sort out the diagnosis. Yet, there is an increasing body of knowledge which presents information casting concern on the adequacy of these measurements. In addition, recent data suggest that chest physicians, occupational medicine physicians, and allergists most often make a diagnosis of occupational asthma by usual clinical methodology, which is a routine part of their hospital's pulmonary function laboratory. SUMMARY: The apparent lack of training in the approach to specific inhalation challenge testing in fellowship programs, the relatively few specialized centers, and the apparent lack of recognition by many physicians who are presented with a patient with suspected occupational asthma means that the use of routine methods to make the clinical diagnosis may often be incorrect, making it unlikely that this approach to the diagnosis of occupational asthma will change in the near future. PMID- 12750607 TI - Costs of occupational asthma and of occupational chronic obstructive pulmonary disease. AB - PURPOSE OF REVIEW: The purpose of this review is to summarize the most recent findings on the costs of occupational asthma and chronic obstructive pulmonary disease, illustrating the different approaches for estimates, the results and the social and financial implications. RECENT FINDINGS: Many published data in the literature show that occupational asthma and chronic obstructive pulmonary disease represent a relevant proportion of the total prevalence of these diseases. Previous findings demonstrated that occupational asthma has adverse economic and employment consequences for the worker, and a significant cost related to the claims. Recent studies using a standard method have specifically estimated the cost of the proportion of cases of asthma and chronic obstructive pulmonary disease attributable to occupational exposure out of the total costs of these diseases, and demonstrated that it is significant. Some recent data indicate a beneficial outcome of occupational asthma and a reduction in the number of the claims from primary and secondary preventive intervention at the workplace. SUMMARY: In conclusion, occupational asthma and chronic obstructive pulmonary disease are likely to pay a substantial contribution to the total costs of the two disorders. These costs are expected to rise with the increasing prevalence of the diseases. Intervention strategies for effective control and prevention at the workplace should lessen the burden of long-term illness and impact on public health costs. PMID- 12750608 TI - Occupational dermatoses. AB - PURPOSE OF REVIEW: All terms referring to job-related (occupational) rashes are clearly defined. Ranking as the second most common occupational disorder, the importance of occupational dermatoses is illustrated. RECENT FINDINGS: 90-95% of occupational dermatoses result from a contact-type dermatitis. Until recently, 4/5 cases were believed to be of the irritant, rather than allergic type of contact dermatitis, however, with more extensive patch testing, more than half the cases were found to be allergic. SUMMARY: The direct causes of occupational dermatoses: chemical, mechanical, physical, and biological in nature are presented. The implications of assigning the degree and type of occupational dermatoses are explained. Workmans compensation is based on a no-fault arrangement, stating that unrestricted treatment and compensation is allowable only when negligence on the part of the employer is proven. Employee benefits are then determined by the duration and degree of disability sustained - all of which becomes litigational. Predisposing host factors, i.e. intellectual ability, anatomic site, atopy, skin pigmentation, age of skin, and immunosuppression (by disease or medication) are usually ignored. Environmental factors at the workplace must be considered conducive to occupational diseases. The full spectrum of irritant contact dermatitis is discussed with examples of each described. Allergic contact dermatitis and patch testing - the gold standard for identification of putative allergens is extensively discussed. Differentiating irritant contact dermatitis from allergic contact dermatitis is elucidated. The other non-contact type skin reactions are briefly mentioned. PMID- 12750609 TI - Early origins of allergic disease: a review of processes and influences during early immune development. AB - PURPOSE OF REVIEW: With the disturbing increase in allergic disease, there is a pressing need to determine the causes, pathogenesis and safe avenues for disease prevention. Although events in early life appear important, no causal pathways have been identified. This review examines new developments in the area of foetal and early postnatal immune maturation. It secondly addresses early predisposing influences and protective factors that may have a future role in allergy prevention. RECENT FINDINGS: New developments in the understanding of the ontogeny of allergen-specific immune responses in atopic infants are discussed, including the role of early type 1 and 2 immune responses, and how these are influenced by perinatal antigen presenting cell and T-cell immaturity. The controversial role of early dietary exposures including breastfeeding, food allergens, hydrolyzed formulae and other dietary factors including omega-3 fatty acids are discussed in the context of the most recent literature. Equally contentious, the role of early house dust and pet allergen exposure is discussed in light of new epidemiological studies and disappointing early results of multicentre allergen avoidance studies. Finally, a number of studies in animals and humans suggest that bacterial products can influence early immune development, providing a new potential therapeutic avenue for disease treatment and prevention. SUMMARY: Complex multifactorial genetic and environmental interactions make research in this area difficult and apparent associations with allergic disease may not be causal in nature. Many current targets for prevention, such as early allergen exposure and infant feeding practices, are proving to be ineffective and may not be directly implicated in rising rates of disease. PMID- 12750611 TI - Oral and sublingual immunotherapy in paediatric patients. AB - PURPOSE OF REVIEW: Sublingual immunotherapy is becoming a routine treatment for respiratory allergy in several countries and it has been validated in international documents. This article will review the available literature on oral and sublingual immunotherapy, discussing the possible use of sublingual immunotherapy in paediatric patients. RECENT FINDINGS: As oral immunotherapy was found to be poorly effective in clinical trials, its use has been discontinued. In contrast, several controlled studies have shown the efficacy of sublingual immunotherapy in children with allergic asthma and rhinitis, and a postmarketing survey has confirmed its safety. Moreover, new data on the long-lasting efficacy of this treatment and on the absence of local immunological effects have recently been published. SUMMARY: The clinical efficacy and the optimal safety profile of sublingual immunotherapy make it a good candidate for treating respiratory allergy in children. Some aspects, such as the dose-response relationship and preventive effect, will be a research challenge for future developments and better definition of indications in children. PMID- 12750610 TI - Nitric oxide in allergic airway inflammation. AB - PURPOSE OF REVIEW: Exhaled nitric oxide has been proposed as a useful noninvasive marker of airway inflammation in asthma. Great efforts have been made to standardize the methodology for exhaled nitric oxide measurement in both children and adults. As a consequence there is now an opportunity to establish the precise relationship between exhaled nitric oxide, atopy and airway inflammation, and to investigate whether or not there is a role for the measurement of exhaled nitric oxide in the management of patients with asthma. RECENT FINDINGS: A number of recent studies have investigated the relationship between exhaled nitric oxide and airway inflammation in asthma measured directly, using sputum induction, bronchoalveolar lavage and endobronchial biopsy. These measurements suggest that exhaled nitric oxide reflects eosinophilic airway inflammation in asthma, although there is no evidence for any relationship between exhaled nitric oxide and other airway inflammatory cells. Exhaled nitric oxide levels were found to be higher in atopic compared with nonatopic groups. These levels, however, are further elevated in atopic patients with asthma, suggesting that exhaled nitric oxide is not simply a marker of atopy. Although there is little evidence to support the routine use of measurement of exhaled nitric oxide in the management of patients with asthma, it may prove to be useful in assessing adherence to treatment with inhaled corticosteroids, or in the identification of patients in whom respiratory symptoms are associated with eosinophilic airway inflammation. SUMMARY: There is good evidence that exhaled nitric oxide reflects eosinophilic airway inflammation in asthma. Well designed, long-term studies are needed to evaluate whether the addition of exhaled nitric oxide measurements to clinical and lung function assessment results in improved asthma control. PMID- 12750612 TI - Evaluation of Lossy data compression of chest X-rays: a receiver operating characteristic study. AB - RATIONALE AND OBJECTIVES: To evaluate the quality of chest radiographs after 32:1 compression/decompression with different image compression algorithms. METHODS: Ten digital (Thoravison) radiographs of an anthropomorphic chest phantom with superimposed simulated nodular lesions (NL) and linear reticular lesions (LL) were obtained. Each radiograph was subdivided into 15 fields; they contained the lesions with a probability of 0.5. The radiographs were compressed and decompressed by using JPEG, fractal and wavelet algorithms at a compression rate of 32:1. Five radiologists evaluated the images. Data were analyzed with the receiver operating characteristic (ROC) method (comparison of area under curve). RESULTS: At 32:1 JPEG or wavelet compression, no statistically significant difference was observed for both NL and LL when compared with the original images. The fractal algorithm performed significantly lower for both NL and LL when compared with the original radiographs. CONCLUSION: The JPEG and wavelet image compression does not result in loss of relevant information for chest x rays at a compression rate of 32:1. PMID- 12750614 TI - Application of adaptive image processing technique to real-time spatial compound ultrasound imaging improves image quality. AB - RATIONALE AND OBJECTIVES: To assess the impact of adaptive filter postprocessing on quality of ultrasound images. METHODS: Ultrasound images acquired with real time spatial compound imaging (SonoCT imaging) were subsequently processed with an adaptive real time algorithm (XRES imaging). Conventional and XRES-processed images from abdominal, pediatric or small parts ultrasound explorations were compared. The delineation of borders, tissue contrast, amount of noise, and overall image quality were evaluated. RESULTS: Delineation of borders and tissue contrast were improved on all images (P < 0.05). The amount of noise was reduced (P < 0.05). The overall image quality was improved for abdominal, pediatric and small parts ultrasound explorations (P < 0.05). No image degradation was found. CONCLUSIONS: Adaptive processing provided better image quality without loss of clinically useful information. PMID- 12750613 TI - F-18 fluorodeoxyglucose positron-emission tomography in the diagnosis of tumor recurrence and metastases in the follow-up of patients with breast carcinoma: a comparison to conventional imaging. AB - AIM: To evaluate the role of F-18-fluorodeoxyglucose positron-emission tomography (F-18 FDG PET) in the follow-up of breast carcinoma in case of clinical suspicion of local recurrence or distant metastases and/or tumor marker increase in correlation to conventional imaging. MATERIAL AND METHODS: Retrospective analysis of the results of F-18 FDG PET (ECAT ART(R), Siemens CTI MS) of 62 patients (age 58.5 +/- 12.8) with surgically resected breast carcinoma (time interval after surgery, 86 +/- 82 months, mean follow-up 24 +/- 12.6 months). Patient- and lesion-based comparison with conventional imaging (CI) including mammography (MG), ultrasonography (US), computerized tomography (CT), magnetic resonance imaging (MRI), radiography (XR) and bone scintigraphy (BS). Furthermore, we evaluated the influence on tumor stage and therapeutic strategy. A visual qualitative evaluation of lesions was performed. RESULTS: On a patient base, sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy for detecting local recurrence or distant metastases were calculated to be 97%, 82%, 87%, 96% and 90% compared with 84%, 60%, 73%, 75% and 74% with CI. On a lesion base, significantly more lymph node (84 vs. 23, P < 0.05) and fewer bone metastases (61 vs. 97, P < 0.05) could be detected by using F-18 FDG PET compared with CI. Sclerotic bone lesions were predominantly detected by BS. On the other hand, there were several patients with more FDG positive bone lesions and also mixed FDG positive/Tc-99m methylenediphosphonate (MDP) negative and FDG negative/Tc-99m MDP positive metastases. In case of normal tumor markers, sensitivity, specificity, PPV, NPV and accuracy for detecting local recurrence or distant metastases were calculated to be 100%, 85.0%, 78.6%, 100% and 90.3% for FDG PET and 80%, 50%, 50%, 80% and 61.5% for CI. An upstaging could be observed in 9.7% (6/62) and downstaging in 12.9% (8/62), leading to a change in therapeutic regimen in 13 patients (21%). CONCLUSIONS: F-18 FDG PET demonstrates apparent advantages in the diagnosis of metastases in patients with breast carcinoma, compared with conventional imaging on a patient base. On a lesion base, significantly more lymph node and less bone metastases can be detected by using F-18 FDG PET compared with conventional imaging, including bone scintigraphy. In patients with clinical suspicion but negative tumor marker profile, too, F-18 FDG PET seems to be a reliable imaging tool for detection of tumor recurrence or metastases. Considering the high predictive value of F-18 FDG PET, tumor stage and therapeutic strategy will be reconsidered in several patients. PMID- 12750615 TI - Navigator-gated coronary magnetic resonance angiography using steady-state-free precession: comparison to standard T2-prepared gradient-echo and spiral imaging. AB - RATIONALE AND OBJECTIVES: Recent developments of magnetic resonance imaging enabled free-breathing coronary MRA (cMRA) using steady-state-free-precession (SSFP) for endogenous contrast. The purpose of this study was a systematic comparison of SSFP cMRA with standard T2-prepared gradient-echo and spiral cMRA. METHODS: Navigator-gated free-breathing T2-prepared SSFP-, T2-prepared gradient echo- and T2-prepared spiral cMRA was performed in 18 healthy swine (45-68 kg body-weight). Image quality was investigated subjectively and signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR) and vessel sharpness were compared. RESULTS: SSFP cMRA allowed for high quality cMRA during free breathing with substantial improvements in SNR, CNR and vessel sharpness when compared with standard T2-prepared gradient-echo imaging. Spiral imaging demonstrated the highest SNR while image quality score and vessel definition was best for SSFP imaging. CONCLUSION: Navigator-gated free-breathing T2-prepared SSFP cMRA is a promising new imaging approach for high signal and high contrast imaging of the coronary arteries with improved vessel border definition. PMID- 12750617 TI - Embolization with autologous fibroblast-attached platinum coils in canine carotid artery aneurysms: histopathological differences from plain coil embolization. AB - PURPOSE: This study was designed to evaluate the effect of autologous fibroblast attached platinum coils on the promotion of intra-aneurysmal thrombus organization. MATERIALS AND METHODS: Sidewall aneurysms were surgically created at common carotid arteries bilaterally in 5 dogs. Fibroblasts were obtained from the skins of the dogs and cultured and attached to unmodified platinum coils in a 5-day coculture. In each animal, one aneurysm was embolized with fibroblast attached platinum coils (study group) and the other with plain platinum coils of the same size (control group). RESULTS: All aneurysms were partially occluded; the mean occlusion rate was 67%. The histologic analyses performed 2 weeks after the embolization showed more advanced organization of intra-aneurysmal thrombus in the study group than in the control group (80% versus 53%, P = 0.02). Overgrowth of the organizing tissue and parent artery stenosis did not occur. CONCLUSION: Our results suggest that autologous fibroblasts delivered with platinum coils could promote the organization of intra-aneurysmal thrombus. Although further research is needed, especially on the potential complications, endovascular treatment using autologous fibroblasts may be applied to aneurysm treatment. PMID- 12750616 TI - Local 3D scaling properties for the analysis of trabecular bone extracted from high-resolution magnetic resonance imaging of human trabecular bone: comparison with bone mineral density in the prediction of biomechanical strength in vitro. AB - RATIONALE AND OBJECTIVES: A novel, nonlinear morphologic measure [DeltaP(alpha)] based on local 3D scaling properties was applied to high-resolution magnetic resonance images (HR-MRI) of human trabecular bone to predict biomechanical strength in vitro. METHODS: We extracted DeltaP(alpha) and traditional morphologic parameters (apparent trabecular volume fraction, apparent trabecular separation) from HR-MR images of 32 femoral and 13 spinal bone specimens. Furthermore, bone mineral density (BMD) and maximum compressive strength (MCS) were determined. The morphologic measures were compared with BMD in predicting the biomechanical strength. RESULTS: In the vertebral (femoral) specimens, R2 for MCS versus DeltaP(alpha) was 0.87 (0.61) (P < 0.001). Correlation between BMD and MCS was 0.53 (P = 0.05) (0.79 [P < 0.001]) for the vertebral (femoral) specimens. For the femoral specimens, prediction of MCS could be improved further by combining BMD and morphologic parameters by multiple regression (R2 = 0.88). CONCLUSIONS: Morphologic measures extracted from HR-MRI considering local 3D scaling properties can be used to predict biomechanical properties of bone in vitro. They are superior to 2-dimensional standard linear morphometric measures and, depending on the anatomic location, more reliably predict bone strength as measured by MCS than does BMD. PMID- 12750618 TI - Fast interactive real-time magnetic resonance imaging of cardiac masses using spiral gradient echo and radial steady-state free precession sequences. AB - RATIONALE AND OBJECTIVES: Cardiac and respiratory controlled MR-imaging is the gold standard for imaging of cardiac masses. However, this technique may be limited in patients with dyspnoe or arrhythmia. The aim of this study was the evaluation of an interactive MR-approach for the detection and localization of cardiac masses. METHODS: Interactive real-time spiral gradient-echo (spiralGE) and radial steady-state-free-precession (radialSSFP) MR-imaging was performed during free-breathing and without cardiac triggering in 15 patients with 14 intracardiac or paracardiac masses. Standard cardiac triggered segmented k-space breath-hold steady-state-free-precession cine MR-imaging was used as the reference MR-imaging technique. Two groups of investigators blinded to clinical data were ask to rank image quality and to identify cardiac masses on real-time MR-images. RESULTS: Image quality was superior using radialSSFP when compared with spiralGE. Using radialSSFP all masses were correctly detected while 6 of 14 masses were missed on spiralGE. Mean real-time MR-imaging time was less than 3 minutes for both techniques. CONCLUSION: Interactive real-time radialSSFP MR imaging allows for accurate and fast detection of cardiac masses without the need of cardiac or respiratory triggering. PMID- 12750619 TI - Histologic evolution of high-intensity focused ultrasound in rabbit muscle. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to examine the histologic evolution over time of rabbit skeletal muscle thermally ablated with high intensity focused ultrasound. The objectives included determining the extent and focality of damage created by this noninvasive, transcutaneous ablative technology. METHODS: Transcutaneous, thermal ablation with an external focused ultrasound transducer was applied to the paraspinous muscles of 19 rabbits. At varying times, up to 100 days after therapy, single sonications were examined histologically. RESULTS: Initially, only subtle staining changes were identified within lesions. In the chronic phase (day 51-100), the muscle was replaced or infiltrated by variable amounts of scar and fat similar to degenerative muscle disorders. Histologic changes were limited to the tissue within the intensity focus of the transducer and were not seen in intervening tissues. DISCUSSION: The current study took a systematic approach to study the long term, in vivo histologic effects of single HIFU lesions in a nonregenerative tissue. This experience in muscle tissue will provide a basis for understanding ultrasound effects for clinical applications such as treatment of uterine fibroids, cardiac tissue, and sarcomas. PMID- 12750620 TI - Electrical impedance scanning in classifying suspicious breast lesions. PMID- 12750622 TI - [Protocol for adapting treatment with vitamin K antagonists before dental extraction]. AB - BACKGROUND: The purpose of this study was to validate a protocol for dental extraction in patients taking vitamin K antagonists without changing the treatment when the INR is lower than 2.8. MATERIAL AND METHODS: One hundred four extractions (96 patients) were performed during a 9 month period in patients taking vitamin K antagonists. Extraction was performed when the international normalized ratio (INR) was less than 2.8, otherwise, the treatment was modified until the desired INR was reached. Extractions were performed under para-apical and alveolar local anesthesia and sutured with hemostatic gauze. RESULTS: Three patients developed postoperative bleeding requiring alveolar revision with local application of tranexamic acid and in one case use of a biological glue. DISCUSSION: The following protocol can be proposed: ambulatory extraction under local anesthesia with an INR the day before extraction: the extraction is performed if the INR is<=2.8 using hemostatic gauze suture and tranexamic acid in case of persistent bleeding. PMID- 12750623 TI - [Dental extraction in patients with bleeding disorders. Proposal of a protocol based on the type of anesthesia used]. AB - INTRODUCTION: The authors propose a protocol to avoid bleeding complications in patients with bleeding disorders. MATERIAL AND METHOD: When a general anesthesia or trunk nerve infiltration is indicated, clotting factor concentrates are to be used in patients with severe bleeding disorders. Desmopressin is to be used in patients with mild bleeding disorders but who are good responders and in patients with thrombopathy. An antifibrinolytic treatment and a fibrin glue are also used. Ninety-six patients underwent 107 extractions with this protocol. RESULTS: Only two patients had bleeding complications requiring an additional treatment. DISCUSSION: In case of bleeding disorders, the treatment depends on disease severity and type of anesthesia. A general treatment (clotting factor concentrates or desmopressin) is indicated with general anesthesia and with local anesthesia in severe bleeding disorders, but not absolutely necessary with local anesthesia in mild bleeding disorders. However, desmopressin can used in all good responders. PMID- 12750624 TI - [Prevention of postoperative bleeding in patients taking oral anticoagulants. Effects of tranexamic acid]. AB - INTRODUCTION: We present a prospective clinical study on use of tranexamic acid mouthwashes in patients taking oral anticoagulants and who have to undergo minor ambulatory oral surgery, without modifying their anticoagulant therapy. MATERIAL AND METHODS: Forty patients (18 men and 22 females) aged from 42 to 81 were studied from April 2000 to December 2002. All patients were under the same anticoagulant, vitamin K antagonist, (Sintrom) for different pathologies. The anticoagulant therapy was not modified neither before nor after surgery. More than 70 oral procedures were carried out under local anesthesia. The International Normalized Ratio of prothrombin time (INR) was<=4. Tranexamic acid 5% (Exacyl) mouthwashes were prescribed during and after surgery. RESULTS: All the patients were followed up during 2 weeks after surgery. There were no postoperative bleedings. DISCUSSION: Use of tranexamic acid mouthwashes under a specific protocol is an efficient, reliable and economic method in preventing postoperative bleeding in patients under oral anticoagulant therapy. PMID- 12750625 TI - [Platelet aggregation inhibitors and vitamin K antagonists in stomatology and maxillo-facial surgery]. AB - Anti-thrombin, anti-platelet and anti-vitamin K agents are common treatments given for the prevention of thrombotic or embolic events. In oral or maxillofacial surgery, the question is whether oral anticoagulants or anti platelets should be discontinued or not: temporary interruption could limit the risk of bleeding, but would increase the risk of thrombosis, and inversely. The best risk/benefit ratio must be achieved, but in many cases anti-platelet agents or even anti-vitamin K agents can be continued. In case of "potentially hemorrhagic" oral surgery, anti-platelet agents should be interrupted about ten days before the intervention. Anti-vitamin K agents should be interrupted four days before surgery and if required, replaced by low-molecular weight heparin at preventive or curative doses. In the very large majority of patients however, surgery (avulsion for example) can be performed without interrupting anticoagulation. Data in the literature have demonstrated that despite the theoretical risk of hemorrhage, bleeding is minimal and can be easily controlled by local treatments; the risk of thrombotic or thromboembolic events after discontinuing anticoagulation is much greater. A close collaboration between the dentist, the stomatologist, and the clinician is required to determine whether anti-thrombosis prevention should be continued and whether the anticoagulation level is adapted to the thrombotic risk. Good surgical technique and local anti hemorrhagic treatment are essential in oral surgery, particularly in patients taking anti-platelets or oral anticoagulants. PMID- 12750626 TI - [Routine preoperative evaluation of hemostasis]. AB - Preoperative evaluation of hemostasis is mainly based on the clinical investigation. Clinical symptoms and history taking help target the most at risk patients. Clinical assessment is also helpful in defining the acquired or constitutional nature of any hemostatic disorder and the type of anomaly involved, e.g. primary hemostatic disorder or coagulation defect. Proper evaluation also helps detect the potential risk of bleeding independently of biological defects detectable with routine tests. Applied in an informative population, clinical evaluation enables optimal utilization of laboratory results. PMID- 12750627 TI - [Brief instructions for the use of expertise by stomatologists and maxillofacial surgeons--Part 1. Different kinds of expertise]. PMID- 12750628 TI - [Our physiotherapy treatment of articular fractures of the mandibular condyle]. AB - Physical therapy greatly contributes to improved function of the injured temporomandibular joint, particularly after trauma. In our unit, we use the Delaire rehabilitation method for patients presenting a fracture of the mandibular condyles. This method involves active mobilization, first with assistance, then with facilitation, and finally against resistance. A rehabilitation session starts with a preparation of the teguments and muscles associated with relaxation exercises. The joint is first mobilized by assisted movements if needed. When unassisted motion becomes possible, propulsion, diduction and open-close exercises are then performed with neuromuscular facilitation. When sufficient amplitudes have been achieved, the program proceeds with opposed exercises. By inducing propulsion and diduction (lateral pterygoid muscle) movements, physical therapy stimulates regeneration of the condylar unit, thus facilitating optimal functional rehabilitation. Posture and passive motion methods, which in our opinion are poorly adapted to the temporomandibular joint, are used little in our unit. Since condylar regeneration is controlled by functional factors, the lateral pterygoid muscle is an important element. Good functional outcome, and the absence of ankylosis, depends directly on the quality of active rehabilitation. PMID- 12750629 TI - [Non-Hodgkin's malignant lymhoma of mental soft tissue: a case report]. AB - Non-Hodgkin's malignant lymphoma is rarely located in the soft tissue of the chin making diagnosis difficult. Clinical signs are non-specific: progressive weight loss, asthenia, fever, an inflammatory tumefaction unresponsive to medical treatment. Imaging shows a homogeneous tumefaction of the soft tissues. Pathology and immunohistochemistry establish the diagnosis. PMID- 12750631 TI - [Antifungal agents]. PMID- 12750630 TI - [Primary non-Hodgkin lymphoma of the mandible: apropos of three cases]. AB - BACKGROUND: Primary non-Hodgkin lymphoma is rarely located in bony tissue, observed in only 5% of primary bone tumors and 4-6% of extranodal non-Hodgkin lymphomas. The mandibular localization is exceptional (0.6%). Clinical and histological features may be misleading, raising the risk of late diagnosis. Chemotherapy and radiotherapy are required. CASE REPORTS: We report three cases of primary non-Hodgkin lymphoma of the mandible diagnosed between 1993 and 2000. All patients were males, aged 9, 37, and 45 years. Mean delay to diagnosis was 8 months. The inaugural complaint was a painful mass of the mandible in all patients. The histomorphology study and immunohistochemistry established the diagnosis. Two patients had diffuse large B cell non-Hodgkin lymphoma, the third anaplastic non-Hodgkin lymphoma. All patients were staged IE and given chemotherapy, associated with radiotherapy in one case. All patients achieved complete remission and continued to be in good health during follow-up lasting from 1 to 9 years. PMID- 12750633 TI - [Cancer, spouse, couple: shoot the intruder]. PMID- 12750632 TI - [Surgery of mandibular condyle. Summary]. PMID- 12750635 TI - [Development and repair of vessels from bone marrow stem cells in the adult: from biology to clinical practice]. AB - In addition to its role in hematopoiesis, bone marrow appears to be a reservoir of stem cells that can differentiate into components of vessel wall. Upon stimulation by factors such as tissue ischemia, bone marrow stem cells can enter into circulating blood, and incorporate into areas of vascular development. Animal models suggest that bone marrow is a cellular source for tissue repair and/or regeneration. Data from humans, and clinical trials using bone marrow stem cells for the treatment of chronic ischemia in limbs and myocardium, support the view that stem cells may represent a new tool for the treatment of ischemia. PMID- 12750636 TI - [High-dose external ionizing radiation prevents intimal hyperplasia and limits secretion of growth factors PDGFbb, bFGF, and TGFb1. Experiments and results]. AB - PURPOSE: To evaluate the ionizing radiation for intimal hyperplasia prevention and to assess the production of growth factors. METHODS: An oversized injury using an embolectomy catheter was performed on a rabbit distal aorta (N=23), associated (test group; N=12) or not (control group; N=11) with a post-operative external radiation (25 Gy). At t=45 days, histological studies and morphometric studies were performed on the aorta. Smooth muscular cells and endothelial cells were stained using immuno-histologic revelation. Immuno-histological analysis was performed on arteries for growth factors PDGFbb, bFGF and TGFb1. RESULTS: Twenty one animals survived the procedure, 11 were in the test group and 10 in the control group. Intimal thickness and ratio intima/media were significantly lower after radiation (respectively p=0.008, p=0.008). There was no difference for the medial thickness (p=0.155). Immuno-histochemical positive staining for PDGF and TGFb1 was lower after radiation (respectively 18.44 +/- 2.963% versus 47.64 +/- 6.86%, p<0.001 and 10.11 +/- 3.18% versus 29.45 +/- 4.156%, p<0.001). There was no difference for the expression of bFGF growth factor. After radiation, the media was found to be reduced and replaced by interstitial fibrosis. CONCLUSION: After external radiation the thickness parameter of the intima and the ratio intima/media decreased significantly in comparison with the control group. PDGF and TGFb1 were also less expressed in the artery irradiated. Fibrosis recasting needs to be confirmed by further investigation. PMID- 12750637 TI - [Retrospective study of the one-year patency of a cuffed polytetrafluoroethylene Venaflo-type graft placed for venous hemodialysis access]. AB - PURPOSE: The objective of this study was to determine retrospectively the one year patency of a new polytetrafluoroethylene (ePTFE) graft with a cuff at the graft-vein anastomosis (Venaflo, Bard industries) placed for hemodialysis access. METHODS: From April first 1999 to December thirty first 2001, 37 consecutive patients (56.8 medium age) underwent 39 Venaflo graft for hemodialysis in Vascular Surgery Section from Bordeaux University Hospital. All medical files have been reviewed at the end of the first year following the operation, by consultation or phone call from the referent nephrologist. Six patients deceased with a functional graft before the end of the study. The patency outcomes were calculated with Kaplan Meier life table method. RESULTS: The one year primary patency is 60.03% with a 95% confident interval [43%; 77%] and secondary patency is 83.73% [70%; 96%]. CONCLUSION: The patency of cuffed ePTFE grafts for hemodialysis was satisfactory in comparison with the result of classic ePTFE grafts in the medical literacy. A large prospective multicentric medium term study would be necessary in order to confirm or not these results. PMID- 12750638 TI - [Arterial complications of thoracic outlet syndrome and pseudarthrosis of the clavicle: three patients]. AB - During a 3-year period, three patients developed arterial complications related to congenital or post-traumatic old pseudarthrosis of the clavicle. Arterial complications of pseudarthrosis of the clavicle presenting as a thoracic outlet syndrome are very rare. Symptoms are variable and occur late. Without treatment, the prognosis is poor with spontaneous development of gangrene. Arterial morphology investigations should be undertaken in patients with pseudarthrosis of the clavicle or isolated arterial symptoms involving the upper limb whose radial pulse disappears during postural tests. Duplex Doppler of the subclavian artery is an excellent screening exam but selective arteriography is the gold standard. It shows proximal arterial lesions (embolytic stenosis of the subclavian artery with post-stenotic dilatation), as well as distal embolic complications. Both static and postural tests must be performed to unmask subclavian restriction by the clavicle, proving its causal effect in the arterial complications. There are four clinical varieties: chronic thrombosis of the subclavian artery, distal arterial micro emboli, acute thrombosis of proximal arteries of the upper limb, and subclavian aneurysm. These lesions are thought to be due to chronic constriction and repeated arterial microtrauma. Congenital or post-traumatic pseudarthrosis, hypertrophic callus, arterial restriction by a screw in a clavicular plate, usually explain the arterial lesions. Bone tumors and Paget's disease are potential but exceptional clavicular etiologies. Surgical treatment is always necessary. Clavicular resection is usually needed in case of pseudarthrosis; there is no functional handicap. Plate fixation and autologous grafting, or open reduction and internal fixation are other valid surgical treatments; The embolytic lesions must be treated to prevent recurrence of distal embolization: graft resection and thromboendarteriectomy have been described. Neurological and venous decompression may be associated at the same time. Complementary treatment can be associated: distal bypass, cervicothoracic sympathectomy, in situ thrombolysis or thrombectomy. Endovascular treatment is not indicated. Optimal treatment of clavicular fractures is required to prevent the development of thoracic outlet syndrome. PMID- 12750639 TI - [False anastomotic iliac (prosthesis-prosthesis) aneurysms: treatment with covered endoprostheses]. PMID- 12750640 TI - [Chronic mesenteric ischemia and temporal arteritis]. PMID- 12750641 TI - [Terminology and chronic venous disease]. PMID- 12750642 TI - [Indications for two-level revascularization of the lower limbs]. PMID- 12750654 TI - The course of low back pain in a general population. Results from a 5-year prospective study. AB - OBJECTIVES: To investigate the course of low back pain (LBP) in a general population over 5 years. DESIGN: Prospective population-based survey by postal questionnaires in 1991, 1992, and 1996. SETTING: The municipal of Ebeltoft, Denmark. SUBJECTS: Two thousand people aged 30 to 50 years, representative of the Danish population. Main outcome measure Number of days with low back pain during the past year. RESULTS: One thousand three hundred seventy were recruited of whom 813 (59%) were followed to 5 years. The responders could be divided into 3 groups with regard to LBP: no pain, short-term pain, and long-lasting/recurring pain. More than one third of people who experienced LBP in the previous year did so for >30 days. Forty percent of people with LBP >30 days at baseline remained in that group 1 and 5 years later, and 9% with LBP >30 days in year 0 were pain free in year 5. People with LBP in year 0 were 4 times more likely to have LBP in year 1, and 2 times more likely to be affected in year 5. CONCLUSIONS: Low back pain should not be considered transient and therefore neglected, since the condition rarely seems to be self-limiting but merely presents with periodic attacks and temporary remissions. On the other hand, chronicity as defined solely by the duration of symptoms should not be considered chronic. PMID- 12750655 TI - Somatovisceral response following osteopathic HVLAT: a pilot study on the effect of unilateral lumbosacral high-velocity low-amplitude thrust technique on the cutaneous blood flow in the lower limb. AB - INTRODUCTION: Spinal manipulative treatment is widely used among manual therapists, although knowledge regarding the absolute physiological effects has not been clearly established. In this study, 20 healthy male subjects underwent a unilateral high-velocity low-amplitude thrust (HVLAT) to the lumbosacral junction, while the cutaneous blood flow in the corresponding dermatome of the lower limb was monitored. METHODS: Subjects underwent a sham manipulation before the actual manipulation and acted as their own control. Laser Doppler flowmetry was used to measure relative changes in the cutaneous blood flow over the L5 dermatome for 5 minutes before the sham manipulation, for 5 minutes between the sham and the actual manipulation, and for 5 minutes after the spinal adjustment. Analysis of variance (ANOVA) and Tukey post hoc analysis was used in the interpretation of the data. RESULTS: Twelve nonsmoking subjects, who received a successful HVLAT manipulation, showed a significant increase (P <.001) in blood perfusion, both ipsilaterally and contralaterally. Six smokers responded with a significant decrease in blood flow ipsilaterally (P <.01) and contralaterally (P <.001) after HVLAT manipulation. CONCLUSION: The results from this study support previous published hypotheses that spinal adjustments outside the region of the sympathetic outflow result in an increase in cutaneous blood flow. Further studies will be needed to confirm the outcome of this study, and more knowledge is needed regarding the specific neurophysiological effects of spinal manipulation. PMID- 12750656 TI - Interobserver reliability of the 24-hour schedule in patients with low back pain: a questionnaire measuring the daily use and loading of the spine. AB - BACKGROUND: Low back pain is a major health problem in western industrialized countries. The 24-Hour Schedule (24HS) is an instrument which intends to obtain insight in the use (ie, posture and applied load) of the back. It consists of a questionnaire, a series of photos, and a registration form. OBJECTIVE: To assess the interexaminer reliability of the 24-Hour Schedule in patients with low back pain. Study design Reliability study. METHODS: People with low back pain were included in the study. Sample size calculation indicated that 40 participants would be sufficient to answer the research question. Participants were coded to remain anonymous, and after giving informed consent, they completed a questionnaire. Two trained examiners assessed each participant independently. In total, 5 examiners participated. RESULTS: Forty participants were analyzed. In our study population, the use of the back was approximately 10 times more in a flexed position compared to a lordotic position. Flexed activity was registered in all 80 assessments, but in 39 assessments, there was no registration of any activity in a lordotic posture. In only 1 participant (diagnosed with Bechterew's disease), the use of the back was more in a lordotic than in a flexed posture. The intraclass correlation coefficient of the assessment was 0.81 (95% confidence interval = 0.67-0.89), corresponding with a high level of agreement between the examiners. CONCLUSION: The interobserver reliability of the 24HS appeared to be high. PMID- 12750657 TI - Comparison of dynamic posteroanterior spinal stiffness to plain film radiographic images of lumbar disk height. AB - BACKGROUND: Assessments of spinal stiffness have become more popular in recent years as a noninvasive objective biomechanical means to evaluate the human spine. Studies investigating posteroanterior (PA) forces in spinal stiffness assessment have shown relationships to spinal level, body type, and lumbar extensor muscle activity. Such measures may be important determinants to discriminate between patients with low back pain (LBP) and asymptomatic subjects. OBJECTIVE: To determine the relationships between dynamic PA spinal stiffness and radiographic measures of lower lumbar disk height and disk degeneration. METHODS: L4 and L5 posterior disk height (PDH), vertebral body height (PVH), anterior disk height (ADH), and vertebral body height (AVH) were obtained from digitized plain film anteroposterior (AP) and lateral radiographs of 18 symptomatic LBP patients presenting to a chiropractic office (8 female patients and 10 male patients, aged 15-69 years, mean 44.3, SD 15.4 years). Disk degeneration (DD) and facet arthrosis (FA) were qualitatively assessed from the films by an independent examiner. Anterior disk height ratios (ADHR = ADH/AVH) and posterior disk height ratios (PDHR = PDH/PVH) were calculated from the disk height measurements and were compared to L4 and L5 posteroanterior spinal stiffness obtained using a previously validated mechanical impedance stiffness assessment procedure. RESULTS: One third of the subjects were found to have radiographic evidence of mild or moderate DD and approximately two thirds of the subjects showed signs of mild or moderate FA. The L4 and L5 anterior disk height and posterior disk height were approximately one half and one fifth of the respective vertebral body heights, and the PA stiffness was greater at L4 than at L5. Male subjects had a greater ADHR than female subjects, but female subjects had a greater L4 and L5 PA stiffness in comparison to male subjects; however, these differences were not statistically significant. Posteroanterior L5 vertebral stiffness was found to be significantly correlated to the L5 PDHR. CONCLUSIONS: Computations of spinal input impedance are relatively simple to perform, can provide a noninvasive measure of the dynamic mechanical behavior of the spine, appear to have potential to discriminate pathologic changes to the spine, and warrant further study on a larger sample of normal subjects and patients. PMID- 12750658 TI - Herbal and nutrient supplementation practices of chiropractic patients: an Australian case study. AB - BACKGROUND: Escalating interest in alternative health care has resulted in increased use of nutritional supplements and herbs by consumers. A number of these neutraceuticals interact with prescription drugs and may have adverse effects. OBJECTIVE: This study explored the extent to which chiropractic patients are likely to inform their chiropractic practitioner of their medication practices. METHOD: A case study that included patients attending 21 chiropractic clinics in 5 states and the Australian Capital Territory was undertaken. Seven hundred fifty-eight patients provided information on their nutritional and herbal supplementation practices. Data were collected using a questionnaire that inquired about general life-style and sought particular information about self initiated and professionally recommended medication practices. A list of medications that could influence coagulation was provided. Patients were specifically requested to indicate whether they had informed their chiropractor about any of the medications, herbs, or nutrients they were taking. RESULTS: More than 1 in 3 of the chiropractic patients in this study were taking, either on the advice of a health professional or, more often, on their own initiative, a nutritional supplement and/or herb. Yet, chiropractors were only occasionally informed about the medication practices of their patients. CONCLUSION: In view of the potential health risks and drug interactions, chiropractors should routinely inquire whether their patients are taking herbs or nutritional supplements. PMID- 12750659 TI - Is low back pain part of a general health pattern or is it a separate and distinctive entity? A critical literature review of comorbidity with low back pain. AB - INTRODUCTION: Research concerning the development of low back pain (LBP) has traditionally focused on risk factors in search of explanations. This review focuses on comorbidity as a first step in identifying a frail subpopulation with a higher risk of developing low back pain, in particular persistent low back pain. Research into comorbidity might yield a greater understanding of the underlying mechanism for low back pain. Data sources Medline was searched from the beginning of the data base to December 2000, followed by a search through the authors' personal collections of epidemiologic literature regarding low back pain. All articles written in English were included if they related LBP to at least one other physical disorder. Articles were excluded if the prevalence of such disorders could not be compared to that of a control group or to the expected prevalence in a normal population. Data extraction The retrieved articles were evaluated for quality based on predefined methodological criteria, whereupon information about associations between low back pain and other physical disorders was extracted. RESULTS: Twenty-three articles were included. They showed positive associations to all disorders investigated (headache/migraine, respiratory disorders, cardiovascular disease, general health, and others) with the exception of diabetes. There was very little information regarding temporality, therefore there are no clues as to causal mechanisms. CONCLUSION: The literature leaves no doubt that diseases cluster in some individuals and that low back pain is part of this pattern. However, the nature of the relationship between low back pain and other disorders is still unclear. PMID- 12750660 TI - Chiropractic care of a patient with vertebral subluxation and Bell's palsy. AB - OBJECTIVE: To describe the chiropractic care of a patient medically diagnosed with Bell's palsy and discuss issues clinically relevant to this disorder, such as its epidemiology, etiology, diagnosis, care, and prognosis. CLINICAL FEATURES: A 49-year-old woman with a medical diagnosis of Bell's palsy sought chiropractic care. Her symptoms included right facial paralysis, extreme phonophobia, pain in the right temporomandibular joint (TMJ), and neck pain. Signs of cervical vertebral and TMJ subluxations included edema, tenderness, asymmetry of motion and posture, and malalignment detected from plain film radiographs. INTERVENTION AND OUTCOME: The patient was cared for with full spine contact-specific, high velocity, low-amplitude adjustments (Gonstead Technique) to sites of vertebral and occipital subluxations. The patient's left TMJ was also adjusted. The initial symptomatic response to care was positive, and the patient made continued improvements during the 6 months of care. CONCLUSION: There are indications that patients suffering from Bell's palsy may benefit from a holistic chiropractic approach that not only includes a focus of examination and care of the primary regional areas of complaint (eg, face, TMJ) but also potentially from significant vertebral subluxation concomitants. PMID- 12750661 TI - Principles in integrative chiropractic. PMID- 12750664 TI - Defining the future of orthodontic research. PMID- 12750665 TI - Comparison of peer assessment ratings (PAR) from 1-phase and 2-phase treatment protocols for Class II malocclusions. AB - The purpose of this study was to compare the dentoalveolar outcomes after 1-phase and 2-phase orthodontic treatment of Class II malocclusions. Class II subjects (n = 208) were randomized to 1-phase or 2-phase treatment with either bionator or headgear/biteplate. The peer assessment rating (PAR) was calculated from pretreatment, prephase 2, and final study models. Chi-square, Kruskal-Wallis, and Wilcoxon rank sum tests were used to evaluate the differences among treatment groups, sexes, races, pretreatment, mandibular plane angle, severity, and compliance. Spearman rank correlation coefficients were used to examine relationships between PAR at different times. The dropout rate of 24.6% did not adversely affect the ability to detect differences of clinical importance or impact treatment groups disproportionately. There were no significant differences with respect to initial PAR or final PAR among the 3 treatment protocols. The 2 early treatment groups had lower PAR scores than the 1-phase group before phase 2 (P =.0001). Lower PAR scores were achieved at both the beginning and end of phase 2 in girls (P =.03; P =.02, respectively). There were differences in the pre phase-2 and post-phase-2 PAR scores based on initial severity (P =.0006; P =.02, respectively), with greater improvement in the patients whose malocclusions were less severe initially. Mandibular plane angle had no effect on pre-phase-2 or post-phase-2 PAR scores. These results do not support the hypothesis that different dentoalveolar outcomes are obtained between 2-phase and 1-phase treatment of Class II malocclusions. PMID- 12750666 TI - Factors associated with orthodontists' assessment of difficulty. AB - The difficulty of achieving an ideal or normal occlusion might lie in the pretreatment occlusion, patient-associated factors, and the treatment. The purpose of this study was to identify factors that were related to the treating orthodontist's posttreatment categorization of a case as difficult or easy. Ten orthodontists each identified 10 easy cases and 10 difficult cases that they had treated. The initial malocclusion was measured with the peer assessment rating (PAR) index and the index of orthodontic treatment need (IOTN). Patient and treatment information was obtained from the treatment records. Statistical analysis with parametric or nonparametric testing was performed. Difficult cases had greater severity and need before treatment and greater residual malocclusion and need after treatment. Difficult cases had more chart entries for problems with hygiene and compliance. They were more likely to have had extractions and changes in treatment plan. Difficult cases also required more appointments and a longer treatment duration. Three logistic regression models were developed based on malocclusion severity, patient characteristics, and treatment characteristics. The models support a correlation between greater pretreatment malocclusion severity, 1-phase treatment, and the designation as a difficult case. This study supports a model in which malocclusion severity and factors associated with the patient and the treatment contribute to an orthodontist's categorization of a case as easy or difficult. PMID- 12750667 TI - Clinical outcome monitoring program (COMP): a new application for use in orthodontic audits and research. AB - Awareness has been growing in recent years about the importance of clinical auditing to provide better orthodontic care to patients. A new, flexible software package, Clinical outcomes monitoring program (COMP), designed to facilitate clinical audits and research, is described. COMP is simple and easy to use and allows clinicians to gather a full data set with the PAR, ICON, or IOTN indexes and to accurately describe clinical outcome and activity. An audit was carried out to demonstrate the use of the software. A total of 205 consecutively finished patients from Good Hope Hospital, Sutton Coldfield, United Kingdom, were analyzed with the software. General and personal trends for 2 clinicians working in a multioperator unit were examined. Results for the unit showed an average PAR improvement of 77.5%, with clinician A performing marginally better overall than clinician B. A general tendency to be less efficient at improving the right, rather than the left, molar relationship was identified. This problem was particularly evident for clinician B, who achieved on average only a 10% improvement in this area. We conclude that COMP is potentially a useful and powerful tool to facilitate orthodontic audits and research. PMID- 12750669 TI - Assessment of facial asymmetries from posteroanterior cephalograms: validity of reference lines. AB - The goal of this study was to determine the ability of various horizontal and vertical reference lines to provide measurements of dentofacial asymmetries from posteroanterior (PA) cephalograms. Ten horizontal and 15 vertical reference lines, including best-fit lines and lines most commonly used in PA analysis, were tested. A model of a dry skull was devised to create 30 asymmetric positions of the maxillomandibular complex. The true transverse and vertical asymmetries were calculated based on measurements of changes in the position of 24 dental and skeletal landmarks. A PA cephalogram was obtained for each asymmetric position. The horizontal and vertical reference lines were constructed on each PA cephalogram, and measurements of transverse and vertical asymmetries were obtained relative to the individual reference lines. Linear regression analyses were used to compare the actual asymmetries with those measured cephalometrically, relative to the individual reference lines. The adjusted R(2) values for all 10 horizontal lines indicated excellent agreement between the true asymmetries and the measured vertical asymmetries. Ten vertical lines accurately represented transverse asymmetry. Vertical lines constructed between 2 midline points, with 1 point located on the lower part of the skull, were not valid. The best-fit line and all lines constructed as perpendiculars through midpoints between pairs of orbital landmarks showed excellent validity. Crista galli anterior nasal spine and nasion-anterior nasal spine had the lowest validity and should not be used in cephalometric analysis of asymmetries. The position of anterior nasal spine will be altered in facial asymmetry involving the maxilla. PMID- 12750670 TI - Correlation of replicating cells and osteogenesis in the glenoid fossa during stepwise advancement. AB - The purposes of this study were to quantify the number of replicating mesenchymal cells and to correlate it with the amount of new bone formed in the glenoid fossa during stepwise advancement. We randomly divided 250 female Sprague-Dawley rats, 35 days old, into 10 control groups (n = 5) and 20 experimental groups (n = 10). Fifty rats from the stepwise experimental group received initial advancement of 2 mm and another 1.5 mm of advancement on day 30 by the addition of veeners. On days 3, 7, 14, 21, 30, 33, 37, 44, 51, and 60, the rats were killed. One hour before that, the rats were injected with bromodeoxyuridine (BrdU) intravenously. We cut 7-microm tissue sections through the glenoid fossa sagittally and stained them with anti-BrdU antibody to evaluate the number of replicating mesenchymal cells. During the first advancement, the number of replicating cells in the posterior region of the glenoid fossa showed a significant increase compared with natural growth, but a significant decrease compared with 1-step advancement. On the second advancement, however, an increase in the number of replicating cells was observed on day 37 with a subsequent and significant increase in bone formation on day 44. Mandibular advancement conducted in a stepwise fashion increases the number of replicating mesenchymal cells in the glenoid fossa. However, a minimum threshold of strain must first be exceeded before these mesenchymal cells can differentiate to ultimately form new bone. PMID- 12750671 TI - Proliferation of epithelial rests of Malassez during experimental tooth movement. AB - The epithelial rests of Malassez (ERM), remnants of Hertwig's epithelial root sheath, are found near the root surface in the periodontal ligament. The functional significance of the ERM is still unknown. The purpose of this study was to examine the behavior of the ERM during experimental tooth movement. Tooth movement was achieved in 12 male Sprague-Dawley rats (each, 120-200 g) by placing elastic bands between the maxillary right first and second molars. The left molars served as controls. The rats were killed after 6, 12, 18, 24, 60, and 72 hours. The mitotic activity of the ERM was assessed by injecting the animals with 5-bromo-2'-deoxyuridine (BrdU) 2 to 3 hours before killing by intracardial perfusion with 4% paraformaldehyde. The molar-bearing segments were dissected and processed for histological examination. The incorporated BrdU was detected by immunohistochemistry. The number of cells in each ERM cluster was counted in all groups. In the 18-, 24-, 60-, and 72-hour experimental groups, the cell numbers were significantly higher than in the controls. The surface areas of the ERM clusters were also measured in all groups, but only in the 18-, 24-, 60-, and 72 hour specimens were the areas significantly higher in the experimental than in the control groups. The ERM cells in the experimental specimens were labeled with anti-BrdU, while those in the controls were not. It was concluded that experimental tooth movement stimulates ERM cells to proliferate and increase in size. These increased activities of the ERM are consistent with a putative role for these cells in collagen turnover in the periodontal ligament that is accelerated during tooth movement. PMID- 12750672 TI - Bond failure patterns in vivo. AB - The aim of this study was to identify the presence and pattern of differences in bond failure between tooth types in vivo when bonding orthodontic brackets with the no-mix orthodontic composite adhesive Right-On. In vivo bond failure for a single operator was recorded for 108 consecutive patients undergoing fixed appliance orthodontic treatment. The bond failure data were analyzed by survival analysis. Time to first failure or censorship was recorded for each bonded attachment. Overall failure in the sample matched previous clinical studies but conflicted with previous ex vivo bond strength data. Mandibular and posterior teeth had significantly higher rates of failure than did maxillary and anterior teeth. The type of attachment used had a significant effect on bond survival. The results of this study confirm that in vivo bond survival is not uniform for all teeth. Comparisons between the findings of this study and those of a previous ex vivo study by the same authors failed to validate ex vivo bond strength testing as clinically relevant. PMID- 12750673 TI - Influence of various surface-conditioning methods on the bond strength of metal brackets to ceramic surfaces. AB - With the increase in adult orthodontic treatment comes the need to find a reliable method for bonding orthodontic brackets onto metal or ceramic crowns and fixed partial dentures. In this study, shear bond strength and surface roughness tests were used to examine the effect of 4 different surface conditioning methods: fine diamond bur, sandblasting, 5% hydrofluoric acid, and silica coating for bonding metal brackets to ceramic surfaces of feldspathic porcelain. Sandblasting and hydrofluoric acid were further tested after silane application. A total of 120 ceramic disc samples were produced, and 50 were used for surface roughness measurements. The glazed ceramic surfaces were used as controls. Metal brackets were bonded to the ceramic substrates with a self-curing composite. The samples were stored in 0.9% NaCl solution for 24 hours and then thermocycled (5000 times, 5 degrees C to 55 degrees C, 30 seconds). Shear bond tests were performed with a universal testing device, and the results were statistically analyzed. Chemical surface conditioning with either hydrofluoric acid (4.3 microm) or silicatization (4.4 microm) resulted in significantly lower surface roughness than mechanical conditioning (9.3 microm, diamond bur; 9.7 microm, sandblasting) (P <.001). The surface roughness values reflect the mean peak-and valley distances. The bond strengths of the brackets bonded to the ceramic surfaces treated by hydrofluoric acid with and without silane (12.2 and 14.7 MPa, respectively), silicatization (14.9 MPa), and sandblasting with silane (15.8 MPa) were significantly higher (P <.001) than those treated by mechanical roughening with fine diamond burs (1.6 MPa) or sandblasting (2.8 MPa). The highest bond strength values were obtained with sandblasting and silicatization with silane or hydrofluoric acid without silane; these fulfilled the required threshold. The use of silane after hydrofluoric acid etching did not increase the bond strength. Diamond roughening and sandblasting showed the highest surface roughness; they can damage the ceramic surface. Acid etching gave acceptable results for clinical use, but the health risks should be considered. The silicatization technique has the potential to replace the other methods; yet cohesive failures were observed in the ceramic during removal of the brackets. PMID- 12750674 TI - The effect of blood contamination on the shear bond strength of orthodontic brackets with the use of a new self-etch primer. AB - The success of bonding brackets to enamel with resin bonding systems is negatively affected by contamination with oral fluids such as blood and saliva. The new self-etch primer systems combine conditioning and priming agents into a single application, making the procedure more cost-effective. The purpose of this study was to assess the effect of blood contamination on the shear bond strength of orthodontic brackets with a new self-etch primer. The brackets (precoated brackets APC II, 3M Unitek, Monrovia, Calif) were bonded to 52 extracted human molars according to 1 of 4 protocols. Group I: A self-etch acidic primer, Angel I (3M/ESPE, St. Paul, Minn), was placed on the enamel for 15 seconds, gently dried with air, and light cured for 10 seconds. Brackets were bonded and light cured for 20 seconds. Group II: The enamel surface was contaminated with human blood for 10 seconds, and the blood was blown off with an air syringe for 5 seconds. Angel I was applied on the contaminated surface; the brackets were then bonded and light cured as in group I. Group III: Angel I was applied as described in group I. The surface was then contaminated with human blood for 10 seconds, and the blood was blown off with an air syringe for 5 seconds. Precoated brackets were then bonded as in groups I and II. Group IV: The enamel surface was contaminated with human blood for 10 seconds and blown off with an air syringe for 5 seconds. The self-etch primer, Angel I, was applied as described earlier, and the surface was recontaminated with human blood for 10 seconds, and the blood was blown off with an air syringe for 5 seconds. The brackets were then bonded as in groups I, II, and III. The results of the analysis of variance (F = 13.31) indicated that the shear bond strengths of the 4 groups were significantly different (P =.001). Tukey HSD posterior tests indicated that the uncontaminated control group had a significantly stronger (mean = 6.0 +/- 3.5 megapascal [MPa]) shear bond strength than the groups that had blood contamination, regardless of whether this occurred before (mean = 2.7 +/- 2.4 MPa), after (mean = 1.1 +/- 0.6 MPa), or before and after (mean = 0.5 +/- 0.2 MPa) the application of the self etch primer. Blood contamination at any stage of the bonding procedure results in a significant and drastic drop in the shear bond strength of orthodontic brackets. PMID- 12750675 TI - Tensile bond strength of ceramic brackets bonded to porcelain facets. AB - The number of adults requesting orthodontic treatment is constantly growing, with a resultant increase in the need to bond brackets to porcelain restorations. Because adults often favor more esthetic ceramic brackets, it is important to evaluate the bond strength of ceramic to porcelain restorations and the integrity of the porcelain surface after debonding. Eighty porcelain facets, resembling mandibular incisors, were used. The surfaces were conditioned by either hydrofluoric acid or microetching. Mandibular incisor ceramic brackets were bonded with 1 of 2 adhesives: Ideal 1 without silane or Right-On with silane. Debonding was performed with a tensile force on a universal testing machine. The mode of bond failure was determined macroscopically, and the integrity of the porcelain facets was evaluated microscopically. The results showed that the type of porcelain conditioning technique affects the tensile bond strength of both adhesives. Conditioning with hydrofluoric acid resulted in significantly higher bond strength (Ideal 1, 7.7 +/- 3.0; Right-On, 7.1 +/- 2.6) than conditioning by microetching (Ideal 1, 4.1 +/- 2.3; Right-On, 3.8 +/- 2.4; P <.05). The mode of bond failure was mainly adhesive, and the porcelain facets were not damaged during debonding. This study shows that both Ideal 1 and Right-On are suitable for bonding ceramic brackets to porcelain restorations, if the porcelain has been conditioned with hydrofluoric acid. PMID- 12750677 TI - Incisor crown shape and crowding. AB - The aim of this study was to investigate correlations between the shape of mandibular incisor crowns and crowding. Study models of 50 untreated white subjects (25 men and 25 women) with varying degrees of crowding were studied. The lower incisors were sectioned and imaged at the contact point and midpoint levels, and the mesiodistal width was measured. Crowding was quantified by using both Little's irregularity index and anterior tooth size-arch length discrepancy. The ratio of the 2 mesiodistal widths was correlated with Little's irregularity index and anterior tooth size-arch length discrepancy. In the women, the ratio showed correlations of 0.52 (P =.008) and 0.55 (P =.005) for central and lateral incisors, respectively, with anterior tooth size-arch length discrepancy. No predictors of lower incisor crowding could be established from mandibular incisor crown shape in this study. PMID- 12750678 TI - On the use of personality characteristics in predicting compliance in orthodontic practice. PMID- 12750676 TI - Effects of high-speed curing devices on shear bond strength and microleakage of orthodontic brackets. AB - This study evaluated the shear-peel bond strength and mode of bond failure of 3 curing devices (plasma arc light, argon laser, and conventional halogen light) and 2 orthodontic bracket adhesives with different filler contents (Transbond XT and Adhesive Precoated [APC]). Observations of microleakage were also reported. Ninety human adolescent premolars were randomly divided into 6 groups, and standardized brackets were bonded according to the manufacturers' recommendations. The plasma arc light produced significantly (P =.006) higher bond strength than did the halogen light or the argon laser when Transbond was used. When APC was used, the plasma arc light and the halogen light produced similar results, and they both produced significantly (P =.015) higher bond strengths than did the argon laser. Overall, the APC showed substantially less variation in bond strength than did the Transbond. Although all curing methods showed significant microleakage (P <.001), differences among the 3 curing lights occurred only when APC was used. Microscopic evaluations demonstrated that 95% of the specimens failed for adhesion at the bracket or tooth surface; the argon laser produced the highest adhesive remnant index scores. On the basis of bond strength and microleakage results, the plasma arc light was comparable with or superior to the other curing devices, depending on the adhesive used. PMID- 12750679 TI - Facemask therapy with rigid anchorage in a patient with maxillary hypoplasia and severe oligodontia. AB - In this report, we describe the treatment of a 10-year-old girl with a Class III skeletal relationship with maxillary hypoplasia and severe oligodontia. The maxillary arch was in a complete crossbite relationship with the mandibular arch. The treatment plan called for displacing the maxillary complex anteriorly with a facemask. Because of the lack of available teeth, a rigid anchorage implant was used in combination with the remaining teeth to provide anchorage. A titanium lag screw was placed in the maxillary alveolus. Three weeks later, 800 g of orthodontic force was applied. A significant anterior displacement of the nasomaxillary complex was achieved with the facemask. At the end of treatment, a temporary removable partial denture was placed. PMID- 12750680 TI - Automated custom-manufacturing technology in orthodontics. PMID- 12750681 TI - Litigation, legislation, and ethics: A road well traveled. PMID- 12750683 TI - Ugly turf issues once again? PMID- 12750684 TI - Medical Board of California withdraws petition to the Supreme Court. PMID- 12750685 TI - The furor over alternative therapy for menopause. PMID- 12750687 TI - The Commission for Case Manager Certification's 10th anniversary. PMID- 12750688 TI - CMSA updates standards of practice for case management. PMID- 12750689 TI - How prepared are you? Ethical and legal challenges facing case managers today (CEU). PMID- 12750690 TI - A complex sense of advocacy: the challenges of contemporary discharge planning. PMID- 12750691 TI - Critical thinking and case management. PMID- 12750692 TI - Successful peripheral blood stem cell mobilisation with filgrastim in patients with chronic myeloid leukaemia achieving complete cytogenetic response with imatinib, without increasing disease burden as measured by quantitative real-time PCR. AB - Imatinib mesylate (Glivec) is a selective inhibitor of bcr-abl tyrosine kinase, the product of the Philadelphia chromosome, which is the hallmark of chronic myeloid leukaemia (CML). With imatinib, complete cytogenetic response (CCR) can be achieved in over 70% of newly diagnosed patients with CML. However, the optimal long-term management of patients who achieve CCR after imatinib is unknown. With longer follow-up, it is anticipated that some patients are likely to progress and become candidates for autologous transplantation. We studied filgrastim (r-metHuG-CSF) mobilisation of peripheral blood stem cells (PBSC) in 32 patients who have achieved CCR with imatinib. Our data demonstrate that (1) the target CD34(+) cell yields of >/=2.0 x 10(6)/kg were attained with filgrastim 10 microg/kg/day, in 9/18 (50%) of patients during uninterrupted imatinib therapy, and in 10/14 (70%) when imatinib was temporarily withheld. The median CD34(+) cell yield per aphaeresis was 0.70 x 10(6)/kg (range 0.14-2.18) and 2.90 x 10(6)/kg (range 0.15-8.71) in the two groups, respectively (P&<0.005). (2) The cell yields did not correlate with the duration of imatinib administration. (3) There was no impact of the mobilisation procedure on the level of leukaemia as measured by serial blood bcr-abl levels using real-time quantitative PCR with either protocol. (4) bcr-abl remained detectable at low levels in the harvests in most but not all patients. In conclusion, filgrastim can safely be used to mobilise PBSC in patients who have achieved CCR with imatinib, but CD34(+) cell yields are significantly improved when imatinib is temporarily withheld. PMID- 12750693 TI - Resistance of Philadelphia-chromosome positive leukemia towards the kinase inhibitor imatinib (STI571, Glivec): a targeted oncoprotein strikes back. AB - Cancer research within the last decades elucidated signaling pathways and identified genes and proteins that lead or contribute to malignant transformation of a cell. Discovery of the Bcr-Abl oncoprotein as the molecular abnormality causing chronic myeloid leukemia (CML) paved the way for the development of a targeted anticancer therapy. The substantial activity of imatinib mesylate (STI571, Glivec) in CML and Philadelphia (Ph)-chromosome positive acute lymphoblastic leukemia (Ph+ ALL) changed the therapeutic approach to Ph+ leukemia and rang the bell for a new era of anticancer treatment. However, when the phenomenon of relapse occurred despite continued imatinib treatment, we had to learn the lesson that imatinib can select for a resistant disease clone. If such a clone still depends on Bcr-Abl, it either carries a BCR-ABL point mutation that prevents binding of the drug or expresses the fusion protein at high levels. Alternatively, leukemia cells that harbor secondary genetic alterations resulting in Bcr-Abl-independent proliferation are selected for their growth advantage in the presence of imatinib. Point mutations in the BCR-ABL kinase domain prevent binding of imatinib but still allow binding of ATP, thus retaining Bcr-Abl kinase activity. Mutated BCR-ABL is frequently detected in cases of imatinib-resistant Ph+ leukemia and therefore represents the main challenge for the investigation of alternative strategies to either overcome resistance or to prevent the emergence of a resistant leukemic clone. PMID- 12750694 TI - Prognostic implications of epigenetic silencing of p15INK4B in acute promyelocytic leukemia. PMID- 12750695 TI - Treatment-related mortality and graft-versus-leukemia activity after allogeneic stem cell transplantation for chronic lymphocytic leukemia using intensity reduced conditioning. AB - Allogeneic stem cell transplantation (SCT) using reduced-intensity conditioning (RIC) has potential to be a promising treatment of aggressive chronic lymphocytic leukemia (CLL). Since available clinical data obtained with this novel approach are very limited, we have performed a survey on this issue. Data of 77 patients were collected from 29 European Group for Blood and Marrow Transplantation centers. Median age was 54 (30-66) years, and the median number of previous chemotherapy regimens was 3 (0-8). HLA-identical sibling donors were used in 81% of the cases. Moderate conditioning regimens (mainly low-dose total body irradiation (TBI) or fludarabine-cyclophosphamide combinations) were administered to 56% of the patients, whereas the remainder received more intense conditioning consisting of fludarabine-busulfan or high-dose melphalan combinations. In 40% of the patients, in vivo T-cell depletion (TCD) with anti-thymocyte globulin or CAMPATH-1H was part of the conditioning regimen. Cumulative treatment-related mortality (TRM) was 18% (95% CI 9; 27) after 12 months. Complete chimerism as well as best response was not achieved immediately post-transplant but took a median of 3 months to develop. The 2-year probability of relapse was 31% (95% CI 18; 44), with no event occurring later than 12 months post transplant in the absence of TCD. With one exception, relapses were not observed after onset of chronic graft-versus-host disease. Event-free and overall survival at 24 months were 56% (95% CI 43; 69) and 72% (95% CI 61; 83), respectively. The median follow up was 18 (1-44) months. Donor lymphocyte infusions or secondary transplants were performed in 19 patients with insufficient disease control and/or incomplete donor chimerism post-transplant, leading to a response in seven patients (37%). Preliminary multivariate analysis identified less than PR at transplant (hazard ratio (HR) 3.5; P&<0.01) and alternative donor (HR 3.1; P=0.02) as significant risk factors for relapse, whereas number of previous regimens >2 (HR 5.4; P=0.03), TBI (HR 2.5; P=0.05), and alternative donor (HR 2.3; P=0.08) were risk factors for survival. We conclude that RIC might favorably influence the outcome after allogeneic SCT for CLL by reducing TRM while preserving graft-versus leukemia activity. PMID- 12750697 TI - Bone marrow transplantation: how important is CD34 cell dose in HLA-identical stem cell transplantation? AB - A recent analysis of Fred Hutchinson Cancer Research Center data has been undertaken to investigate the association of infused CD34 cell dose with various clinical outcomes after HLA-identical transplantation. Separate assessments for unrelated vs related donors and the use of bone marrow or mobilized G-CSF stimulated peripheral blood mononuclear cells (G-PBMC) have been incorporated. The three primary findings are: (1) Higher CD34 dose results in better neutrophil and platelet recovery in all settings. (2) Higher CD34 doses (8 x 10(6)/kg) are associated with the development of more chronic graft-versus-host disease when using related G-PBMC. (3) Higher CD34 dose is correlated with improved survival after unrelated donor bone marrow transplantation. These data suggest that the CD34 content of a graft can have a significant impact on clinical outcome after allogeneic transplantation, but defining an optimal dose is dependent on both the type of donor and the stem cell source. PMID- 12750696 TI - In vitro antiproliferative activity of the farnesyltransferase inhibitor R115777 in hematopoietic progenitors from patients with myelofibrosis with myeloid metaplasia. AB - R115777 is an orally bioavailable farnesyltransferase inhibitor (FTI) that has displayed encouraging activity in patients with acute myeloid leukemia. To determine whether R115777 might exert similar activity in myelofibrosis with myeloid metaplasia (MMM), we evaluated its effects on circulating myeloid progenitor cells from patients with MMM (n=25) using in vitro colony-forming assays. The median R115777 concentrations that inhibited colony formation by 50% were 34 and 2.7 nM for myeloid and megakaryocytic colonies from MMM patients, respectively. Progenitors from normal controls and patients with other myeloproliferative disorders demonstrated similar sensitivity. Since the ras polypeptides are one putative target of FTIs, the potential role of ras effectors was examined by incubating parallel progenitor assays with the phosphatidyl inositol-3 (PI-3) kinase inhibitor LY294002 and the mitogen-activated protein kinase 1 inhibitor PD98059. MMM progenitor colonies (n=7) were highly sensitive to LY294002 but not to PD98059, implying that the PI-3 kinase pathway may be critical for survival and proliferation of these cells. In addition to indicating that MMM progenitors are sensitive to clinically achievable R115777 concentrations in vitro, these results provide a potential explanation for the thrombocytopenia observed with R115777 during the treatment of other hematologic malignancies. PMID- 12750698 TI - The presence of an HLA-identical sibling donor has no impact on outcome of patients with high-risk MDS or secondary AML (sAML) treated with intensive chemotherapy followed by transplantation: results of a prospective study of the EORTC, EBMT, SAKK and GIMEMA Leukemia Groups (EORTC study 06921). AB - This report used the framework of a large European study to investigate the outcome of patients with and without an HLA-identical sibling donor on an intention-to-treat basis. After a common remission-induction and consolidation course, patients with an HLA-identical sibling donor were scheduled for allogeneic transplantation and patients lacking a donor for autologous transplantation. In all, 159 patients alive at 8 weeks from the start of treatment were included in the present analysis. In total, 52 patients had a donor, 65 patients did not have a donor and in 42 patients the availability of a donor was not assessed. Out of 52 patients, 36 (69%) with a donor underwent allogeneic transplantation (28 in CR1). Out of 65 patients, 33 (49%) received an autograft (27 in CR1). The actuarial survival rates at 4 years were 33.3% (s.e. = 6.7%) for patients with a donor and 39.0% (s.e. = 6.5%) for patients without a donor (P = 0.18). Event-free survival rates were 23.1% (s.e. = 6.2%) and 21.5% (s.e. = 5.3%), respectively (P = 0.66). Correction for alternative donor transplants did not substantially alter the survival of the group without a donor. Also, the survival in the various cytogenetic risk groups was not significantly different when comparing the donor vs the no-donor group. This analysis shows that patients with high-risk myelodysplastic syndrome and secondary acute myeloid leukemia may benefit from both allogeneic and autologous transplantation. We were unable to demonstrate a survival advantage for patients with a donor compared to patients without a donor. PMID- 12750699 TI - Higher doses of CD34+ peripheral blood stem cells are associated with increased mortality from chronic graft-versus-host disease after allogeneic HLA-identical sibling transplantation. AB - Allogeneic peripheral blood stem cell transplantation (PBSCT) has emerged as an alternative to bone marrow transplantation. PBSCT can be associated with a higher incidence of chronic graft-versus-host disease (cGVHD). In this study, we investigated whether there was a correlation between the composition of PBSC grafts (CD34+ and CD3+ cells) and hematological recovery, GVHD, relapse, and relapse-free survival (RFS) after myeloablative HLA-identical sibling PBSCT. The evolution of 100 acute or chronic leukemia patients was analyzed. Neither hematological recovery, acute or cGVHD, nor relapse, was significantly associated with CD3+ cell dose. Increasing CD34+ stem cells was associated with faster neutrophil (P=0.03) and platelet (P=0.007) recovery. Moreover, 47 of the 78 patients evaluable for cGVHD (60%; 95% CI, 49-71%) developed extensive cGVHD. The probability of extensive cGVHD at 4 years was 34% (95% CI, 21-47%) in patients receiving a 'low' CD34+ cell dose (<8.3 x 10(6)/kg), as compared to 62% (95% CI, 48-76%) in patients receiving a 'high' CD34+ cell dose (>8.3 x 10(6)/kg) (P=0.01). At a median follow-up of 59 months, this has not translated into a difference in relapse. In patients evaluable for cGVHD, RFS was significantly higher in patients receiving a 'low' CD34+ cell dose as compared to those receiving a 'high' CD34+ cell dose (P=0.04). This difference was mainly because of a significantly higher cGVHD-associated mortality (P=0.01). Efforts to accelerate engraftment by increasing CD34+ cell dose must be counterbalanced with the risk of detrimental cGVHD. PMID- 12750700 TI - Cryptic insertion and translocation or nondividing leukemic cells disclosed by FISH analysis in infant acute leukemia with discrepant molecular and cytogenetic findings. AB - Of 51 infants with acute leukemia, 13 (25%) had contradictory findings on 11q23/MLL rearrangements that were analyzed by cytogenetic and Southern blot methods: seven had rearranged MLL and normal karyotype, four had rearranged MLL and abnormal karyotype with no 11q23 translocation, and two had germline MLL and 11q23 translocations. Fluorescent in situ hybridization (FISH) analysis using an MLL probe that was performed to elucidate the discrepancy disclosed the presence of normal dividing cells and nondividing leukemic cells in the same bone marrow in five patients, and cryptic insertion or translocation in another five. Subsequent FISH and reverse transcription-polymerase chain reaction analysis identified the MLL-AF10, MLL-AF4, or MLL-AF1q fusions that were produced by the cryptic rearrangements in four of the five patients. In the remaining three patients, the breakpoint of 11q23 translocation was located distal to the MLL locus in one, and the discrepancy was unresolved in two. Thus, FISH should complement cytogenetic analysis when cytogenetic and molecular genetic findings are contradictory in infant leukemia, and when infant leukemia does not show 11q23 translocations or other specific translocations including t(7;12), t(1;22), etc that are recurrently found in infant leukemia. PMID- 12750701 TI - FLT3-TKD mutation in childhood acute myeloid leukemia. AB - Mutations of receptor tyrosine kinases are implicated in the constitutive activation and development of human hematologic malignancies. An internal tandem duplication (ITD) of the juxtamembrane domain-coding sequence of the FLT3 gene (FLT3-ITD) is found in 20-25% of adult acute myeloid leukemia (AML) and at a lower frequency in childhood AML. FLT3-ITD is associated with leukocytosis and a poor prognosis, especially in patients with normal karyotype. Recently, there have been three reports on point mutations at codon 835 of the FLT3 gene (D835 mutations) in adult AML. These mutations are located in the activation loop of the second tyrosine kinase domain (TKD) of FLT3 (FLT3-TKD). The clinical and prognostic relevance of the TKD mutations is less clear. To the best of our knowledge, there has been no report to describe FLT3-TKD mutations in childhood AML. In this pediatric series, FLT3-TKD mutations occurred in three of 91 patients (3.3%), an incidence significantly lower than that of FLT3-ITD (14 of 91 patients, 15.4%) in the same cohort of patients. None of them had both FLT3-TKD and FLT3-ITD mutations. Sequence analysis showed one each of D835 Y, D835 V, and D835 H. Of the three patients carrying FLT3-TKD, two had AML-M3 with one each of L- and V-type PML-RARalpha, and another one had AML-M2 with AML1-ETO. None of our patients with FLT3-TKD had leukocytosis at diagnosis. At bone marrow relapse, one of the four patients examined acquired FLT3-ITD mutation and none gained FLT3-TKD mutation. PMID- 12750702 TI - Expression of HOX11 in childhood T-lineage acute lymphoblastic leukaemia can occur in the absence of cytogenetic aberration at 10q24: a study from the Children's Cancer Group (CCG). AB - Clonal genetic aberrations in tumour cells provide critical information for the development of new diagnostic and therapeutic strategies for patients. In paediatric T-cell acute lymphoblastic leukaemia (T-ALL) chromosomal translocations are present in 30-35% of cases. HOX11 and the closely related HOX11L2 genes play a key role in T-ALL. HOX11 is aberrantly activated by either of the two chromosomal translocations, t(7;10) and t(10;14). In this study, HOX11 expression levels were measured by real-time quantitative reverse-transcriptase polymerase chain reaction. We show that leukaemic blasts from 15/76 (19.7%) paediatric T-ALL patients expressed the HOX11 gene at high level and 22/76 (28.9%) at low level, yet the reported frequency for chromosomal rearrangement of 10q24 is 4-7%. Direct cytogenetic analysis revealed that only 2/16 specimens that showed HOX11 expression exhibited abnor-malities at 10q24. These results confirm and extend our previously published findings, and implicate mechanisms other than gross chromosomal translocations for the deregulation of HOX11. Analysis of clinical outcome for the whole study group showed a trend for better outcome for patients with leukaemic blasts expressing HOX11 at high level. A statistically significant difference in clinical outcome was found in a subgroup of 20 patients treated for high-risk disease on CCG-1901 from the Children's Cancer Group, where HOX11 expression in leukaemic blasts conferred a prognostic advantage (P=0.01). PMID- 12750703 TI - Dendritic cells loaded with apoptotic tumour cells induce a stronger T-cell response than dendritic cell-tumour hybrids in B-CLL. AB - Dendritic cells (DC) are professional (specialised) antigen-presenting cells that can capture antigen from apoptotic tumour cells and induce MHC class I- and II restricted responses. Also, DC fused with tumour cells may be effective for immune response induction. Both cell preparations may be considered as vaccine candidates in a therapeutic approach. We examined autologous T-cell activation by DC that had endocytosed leukaemic B-cell apoptotic bodies (Apo-DC) and compared it to the T-cell stimulatory capacity of DC that were fused with tumour cells. Following incubation, 22.6+/-6.2 (mean+/-s.e.m.) of DC had endocytosed leukaemic cells, while the frequency of DC-leukaemic cell hybrids was 10.5+/-2.6%. Apo-DC and hybrid cells both demonstrated the ability to stimulate a tumour-specific T cell immune response in vitro. A T-cell proliferation response was also observed in four out of five CLL patients when using Apo-DC. However, fusion hybrids lacked the ability to elicit a proliferative response. Apo-DC also induced an IFN gamma response, as did hybrid cells. The cytokine response induced by Apo-DC was significantly higher than that induced by fusion (P<0.05). This study shows that endocytosed apoptotic tumour cells induced a significantly stronger T-cell response than DC hybrids; and as such should be a better candidate for vaccine production. PMID- 12750704 TI - Efficient elimination of chronic lymphocytic leukaemia B cells by autologous T cells with a bispecific anti-CD19/anti-CD3 single-chain antibody construct. AB - Recently, we have shown that a novel recombinant bispecific single-chain antibody construct (bscCD19 x CD3), induces highly efficacious lymphoma-directed cytotoxicity mediated by unstimulated peripheral T lymphocytes. Functional analysis of bscCD19 x CD3 has so far been exclusively performed with human B lymphoma cell lines and T cells from healthy donors. Here we analysed the properties of bscCD19 x CD3 using primary B cells and autologous T cells from healthy volunteers or patients with B-cell chronic lymphocytic leukaemia (B-CLL). We show that bscCD19 x CD3 induces T-cell-mediated depletion of nonmalignant B cells in all four cases and depletion of primary lymphoma cells in 22 out of 25 cases. This effect could be observed at low effector-to-target (E:T) ratios and in the majority of cases without additional activation of autologous T cells by IL-2. Even in samples derived from patients heavily pretreated with different chemotherapy regimens, strong cytotoxic effects of bscCD19 x CD3 could be observed. The addition of bscCD19 x CD3 to patients' cells resulted in an upregulation of activation-specific cell surface antigens on autologous T cells and elevated levels of CD95 on lymphoma B cells. Although anti-CD95 antibody CH 11 failed to induce apoptosis in lymphoma cells, we provide evidence that B-CLL cell depletion by bscCD3 x CD3 is mediated at least in part by apoptosis via the caspase pathway. PMID- 12750705 TI - Aberrant methylation and impaired expression of the p15(INK4b) cell cycle regulatory gene in chronic myelomonocytic leukemia (CMML). AB - The important cell cycle regulatory gene p15(INK4b) has been shown to be inactivated in acute myeloid leukemia and myelodysplastic syndrome. Little is known about the expression and epigenetic modification of this gene in chronic myelomonocytic leukemia (CMML) that belongs to the myelodysplastic/myeloproliferative disorders (MDS/MPD) with a high proportion of blastic transformation. Analysis of bone marrow trephines in a series of 33 CMML cases showed an aberrant p15(INK4b) gene methylation in up to 58% of cases. Methylation was analyzed employing different methylation-specific PCR and genomic sequencing protocols. It turned out to be spread over a broad area of the 5' region and exhibited substantial heterogeneity between cases and even in individual patients. The degree of aberrant methylation was correlated with a reduced mRNA as well as reduced protein expression, and was associated with a higher expression of DNA methyltransferase DNMT 3A. We conclude that aberrant gene methylation is a frequent event in CMML that might contribute to the pathogenesis of this MDS/MPD. PMID- 12750706 TI - Hypermethylation of GpG islands in the promoter region of p15(INK4b) in acute promyelocytic leukemia represses p15(INK4b) expression and correlates with poor prognosis. AB - We evaluated the methylation status of p15 gene in a series of 65 patients with newly diagnosed acute promyelocytic leukemia (APL) receiving homogeneous treatment. Moreover, in 32 of them, the methylation status of p15 gene was correlated to the p15 m-RNA expression. In total, 31 patients had no p15 methylation (U group). An abnormal methylation pattern was found in 34 patients: in seven of these patients only methylated DNA was detected (M group), while in the remaining 27 patients (M/U group), both methylated and unmethylated DNA were amplified. Patients from M group showed a higher incidence of relapses and a lower disease-free survival (DSF) with respect to patients from U and M/U groups (29, 64 and 79% at 5 years for M, U/M and U patients, respectively, P=0.03), while p15 methylation had no impact on overall survival. The p15 expression was detectable in all patients with unmethylated DNA, in none of patients with fully methylated DNA and in 60% of patients with partially methylated DNA. The DFS estimate at 5 years for p15-negative patients was significantly lower than that of p15-positive patients (P=0.03). These data confirm that the presence of p15 methylation negatively influences the prognosis of APL, mainly when it represses the p15 gene transcription. PMID- 12750707 TI - Infectious complications in patients with acute promyelocytic leukaemia treated with the AIDA regimen. AB - Infections represent a frequent complication of chemotherapy used for acute myeloid leukaemia (AML) and are associated with important toxicity frequently leading to treatment discontinuation. Acute promyelocytic leukaemia (APL) is a unique AML subset requiring tailored therapy including all-trans retinoic acid and anthracycline-based chemotherapy. We analysed in this study the incidence and type of infections complicating the clinical course of 89 consecutive APL patients receiving the AIDA protocol at a single institution. A total of 179 febrile episodes were registered during induction and consolidation, 52% of which were of unknown origin. Infections were clinically and microbiologically documented in 10.6 and 37.4% of cases, respectively. Coagulase-negative staphylococci represented the major cause of septicaemia (28%) and were more frequently isolated during induction, whereas viridans group streptococci, the second pathogen most frequently isolated from blood (27%), represented the principal pathogen detected during consolidation and were significantly associated with mucositis. Gram-negative bacteria accounted for 33.3% of all blood isolates. Fungal infections were only occasionally observed. Bloodstream infections in APL patients were compared with those documented in 271 consecutive patients affected by other subtypes of AML. The incidence of total septicaemia episodes, of staphylococcal bacteraemias and of fungaemias was significantly higher in patients with other AMLs. Empirical antibiotic therapy with ceftriaxone plus amikacin was effective in 73% of APL cases, most of the remaining cases being successfully managed by the addition of teicoplanin. One single death apparently related to infectious complication was recorded. Overall, infections led to antileukaemic treatment withdrawal in six patients, five of whom currently remain in haematologic remission for 13-106 months. These results indicate that a particular pattern of infections is observed in APL patients receiving ATRA plus anthracycline-based chemotherapy and that these appear to be effectively counteracted by standard management. PMID- 12750708 TI - Glutathione depletion overcomes resistance to arsenic trioxide in arsenic resistant cell lines. AB - Arsenic trioxide (As(2)O(3)) is an effective treatment for acute promyelocytic leukemia (APL), but is less effective against other leukemias. Although the response of APL cells to As(2)O(3) has been linked to degradation of the PML/RARalpha fusion oncoprotein, there is evidence that PML/RARalpha expression is not the only mediator of arsenic sensitivity. Indeed, we found that exogenous expression of PML/RARalpha did not sensitize a non-APL leukemic line to As(2)O(3). To evaluate possible other determinants of sensitivity of leukemic cells to As(2)O(3), we derived two arsenic-resistant NB4 subclones. Despite being approximately 10-fold more resistant to arsenic than their parental cell line, PML/RARalpha protein was still degraded by As(2)O(3) in these cells, providing further evidence that loss of expression of the oncoprotein does not confer arsenic sensitivity. Both arsenic-resistant clones contained high glutathione (GSH) levels, however, and we found that GSH depletion coupled with As(2)O(3) treatment dramatically inhibited their growth. Annexin V-staining and TUNEL analysis confirmed a synergistic induction of apoptosis. In addition, these cells failed to accumulate ROS in response to arsenic treatment, in contrast to their arsenic-sensitive parental cells, unless cotreated with buthionine sulfoximine. While other malignant cells did not show a good correlation between arsenic sensitivity and GSH content, GSH depletion nevertheless sensitized all cell lines examined, regardless of their initial response to arsenic alone. These findings suggest that PML/RARalpha expression is not a determinant of arsenic sensitivity, and further support the coupling of GSH depletion and arsenic treatment as a novel treatment for human malignancies that are unresponsive to arsenic alone. PMID- 12750709 TI - Hematopoietic stem cell transplantation for hematological malignancies in Europe. AB - Hematopoietic stem cell transplants (HSCTs) are considered the best treatment option for many hematological malignancies, and transplant numbers have increased five-fold during the last decade. Only a few controlled prospective studies are available, and different opinions prevail. Data from 118 167 HSCT (36% allogeneic, 64% autologous) collected within the EBMT activity survey from 1990 to 2001 were used to assess trends over time, transplant rates and coefficient of variation (CV) of transplant rates among European countries for acute myeloid leukemia (AML; 18.5%), acute lymphocytic leukemia (ALL; 12%), chronic myeloid leukemia (CML; 11.5%), myelodysplastic syndromes (MDS; 3%), lymphoproliferative disorders (LPS; 36.3%) and multiple myeloma (MM; 18.7%). Transplant rates increased in all countries and for all indications from 1990 to 2001 from 1.7 fold (CML) to 24.8-fold (MM). Transplant rates have declined for CML since 1999. Autologous HSCT are the preferred choice for LPS and MM, allogeneic HSCT for ALL and myeloid malignancies. CVs of less than 50% suggest consensus for allogeneic HSCT in AML, ALL, CML, MDS and NHL, for autologous HSCT in LPS and MM. These data give an overview of the current status of HSCT for hematological malignancies in Europe and provide objective information for health-care providers and patient counselling. PMID- 12750710 TI - Human cord blood long-term engrafting cells are CD34+ CD38-. AB - There have been controversies about CD34 and CD38 expression by human cord blood (CB) stem cells. Using the newborn NOD/SCID/beta2-microglobulin-null mouse assay that we recently developed, we examined the in vivo engrafting capability of human CB cells. Almost all of the 4-5 months engrafting cells were found in CD34(+) population. The capability of secondary reconstitution was found only in the CD34(+) cells. When the CD34(+) CB cells were separated into CD38(-) and CD38(+) subpopulations and tested for engraftment, the majority of the engrafting cells were detected in the CD38(-) subpopulation. These findings are consistent with the results from studies of murine stem cells and strongly indicate that the phenotype of human CB stem cells is CD34(+) CD38(-). PMID- 12750711 TI - An isoform of the Wilms' tumor suppressor gene potentiates granulocytic differentiation. AB - WT1 is expressed in hematopoietic progenitor cells and in acute leukemia, but its role in normal and malignant hematopoiesis has not been clearly defined. Alternative splicing of the WT1 mRNA yields several protein isoforms with distinct DNA binding and transcriptional regulatory activities. In this study, we investigated the effect of the WT1 isoform lacking two alternatively spliced sequences (WT1 (-/-)) in 32D cl3 cells, a murine myeloid progenitor cell line. The expression of WT1 (-/-) accelerated the granulocyte-colony stimulating factor (G-CSF)-mediated differentiation of these cells, as judged by morphology and by the expression of differentiation-associated genes and cell surface antigens. WT1 (-/-) inhibited G1/S progression in G-CSF but not in interleukin-3, potentially accounting for its ability to accelerate differentiation. It is likely that dominant-negative mutants previously reported in leukemia patients participate in leukemogenesis by inhibiting this function of the wild-type protein. PMID- 12750712 TI - Prognostic value of GST-pi expression in diffuse large B-cell lymphomas. AB - Among mechanisms potentially involved in resistance to alkylating agents and anthracyclines, the glutathione system has been extensively studied in vitro. We analyzed by immunohistochemistry the relation between glutathione s-transferase pi (GST-pi) expression in tumor cells and outcome in 69 cases of diffuse large B cell NHL (DLBCL). GST-pi expression was considered as low when <50% of tumor cells were stained and high when >/=50% tumor cells were stained. Median follow up was 58 months. GST-pi expression was correlated with the probability of achieving complete remission (CR). Patients with high GST-pi expression had a worse 5-year freedom from progression (FFP). High GST-pi expression was associated with a trend for lower survival. In the group of patients with International Prognostic Index (IPI) 0-1, low GST-pi expression was associated with a CR rate of 88%, a 5-year FFP of 76+/-20% and a 5-year survival of 78+/-16% compared to 36, 14+/-16 and 40+/-32%, respectively, in patients with a high GST pi expression (P=0.002, P&<10(-5) and P=0.01, respectively). No correlation was found between GST-pi expression and lactico deshydrogenase serum level, age, Ann Arbor stage, performance status, and IPI index. Both GST-pi expression and the IPI index correlated with FFP. After incorporating IPI and GST-pi expression in a multivariate analysis for FFP, GST-p expression remained the only prognostic factor (P=0.003). Our findings suggest that GST-pi expression had strong prognostic significance in DLBCL, which appears to be independent of other prognostic parameters in those disorders. PMID- 12750713 TI - Histological features of acute hepatitis after imatinib mesylate treatment. PMID- 12750714 TI - MLL partial tandem duplications in acute leukemia cell lines. PMID- 12750715 TI - Genotypes of the glutathione S-transferase superfamily do not correlate with outcome of childhood acute lymphoblastic leukemia. PMID- 12750718 TI - A case of acute promyelocytic leukaemia following mitoxantrone treatment of multiple sclerosis. PMID- 12750719 TI - Microdose alpha-interferon shows clinical and antiangiogenic effect in extramedullary myeloid tumor: a case report. PMID- 12750716 TI - Interferon-alpha modulates the immune response enhancing B7-1 and B7-2 costimulatory molecules and T8 lymphocytes in chronic myeloid leukemia. PMID- 12750720 TI - Serum levels of the nm23-H1 protein and their clinical implication in extranodal NK/T-cell lymphoma. PMID- 12750721 TI - Comparison of fludarabine-containing salvage chemotherapy regimens for relapsed/refractory acute myelogenous leukemia. PMID- 12750722 TI - A unique variant of ETV6/AML1 fusion in a child with acute lymphoblastic leukemia. PMID- 12750723 TI - Constitutive phosphorylation of Akt/PKB protein in acute myeloid leukemia: its significance as a prognostic variable. PMID- 12750725 TI - Evolution in alternatives to blood transfusion. PMID- 12750726 TI - Mesenchymal stem cell: use and perspectives. AB - Studies on hematopoiesis have focused on the function and composition of human bone marrow stroma. Stroma function gives hematopoietic stem cells the microenvironment appropriate for self-renewal and/or prompt differentiation into hematopoietic progenitor cells, then into terminal specialized cells. Human bone marrow stroma has been dissected into hematopoietic and nonhematopoietic components. The former includes hematopoietic-derived cells, mainly macrophages, while the latter, still poorly characterized, is composed mainly of endothelial and mesenchymal stem cells and their derivatives (adipocytes, chondrocytes, cells of the osteogenic lineage). Isolation of bone marrow mesenchymal stem cells has made available a population of adherent cells, belonging to the non-hematopoietic stroma, which are morphologically and phenotypically homogeneous. This review will focus on: (i) definition of bone marrow stroma and mesenchymal stem cells; (ii) methods of mesenchymal stem cell isolation, morphological and phenotypic characterization; (iii) mesenchymal stem cell functional and differentiation properties and (iv) therapeutic applications of mesenchymal stem cells. PMID- 12750727 TI - Diagnostic approaches to suspected deep vein thrombosis and pulmonary embolism. AB - The present paper aims at reviewing our present knowledge on the diagnosis of deep vein thrombosis and pulmonary embolism with a particular emphasis on clinically suspected outpatients. The various contemporary diagnostic tools (including clinical probability assessment, fibrin D-Dimer measurement, venous compression ultrasonography, ventilation/perfusion lung scintigraphy, helical CT scan, and echocardiography) are presented and rational sequential strategies are discussed in their various aspects, including validation in outcome studies, and cost-effectiveness analyses. PMID- 12750728 TI - Overexpression of dominant-negative Ikaros 6 protein is restricted to a subset of B common adult acute lymphoblastic leukemias that express high levels of the CD34 antigen. AB - Ikaros is a critical regulator of hematopoiesis. Its effects result in part from the balance between the isoforms that are produced by differential splicing of the pre-mRNA. Short isoforms that lack the DNA-binding domain act as dominant negatives by binding long isoforms through the C-terminal zinc-finger domain, which allows for the homo- or heterodimerization of the proteins. There are a number of evidences that different subsets of murine hematopoietic progenitors - as defined by phenotype - have different patterns of Ikaros expression. Forced expression of short isoforms (Ik5, Ik6 or Ik7) in murine or human hematopoietic progenitors alters the differentiation capacities of these cells. Human leukemias provide additional information: because of the blockade in differentiation, leukemias represent an equivalent of a particular stage of human hematopoietic hierarchy. We and others have shown that human acute leukemias are heterogeneous for the pattern of Ikaros isoform expression. The present study focused on adult de novo B ALLs and the Ikaros 6 isoform. ProB (BI, n=3), common B (BII, n=15) and preB (BIII, n=3) ALL were identified by their phenotype. RT-PCR and Western blot analyses of blast cell protein lysates suggest that approximately 50% of BII leukemias overexpress Ikaros 6 RNA and protein. Comparison of BII cells with high or normal levels of Ik6 shows a higher level of expression for the membrane stem cell antigen CD34 in the former, as detected with flow cytometry and confirmed with DNA arrays. PMID- 12750729 TI - Rituximab in combination with CNOP chemotherapy in patients with previously untreated indolent non-Hodgkin's lymphoma. AB - Rituximab, a chimeric monoclonal antibody, produces response rates of up to 73% in patients with previously untreated indolent non-Hodgkin's lymphoma (NHL), and has high activity when combined with chemotherapy. The purpose of this phase II study was to determine the efficacy and safety of rituximab plus cyclophosphamide, mitoxantrone, vincristine and prednisone (CNOP) chemotherapy in patients with indolent NHL. In all, 42 patients (median age 67 years) with previously untreated follicular, marginal zone or small lymphocytic/lymphoplasmacytic NHL received six infusions of rituximab (375 mg/m(2)) in combination with six cycles of CNOP. The overall response rate was 90% comprising 30 complete (71%) and eight partial (19%) responses. Although patients with marginal zone lymphoma or International Prognostic Index (IPI) score 3 had lower complete response rates, no significant difference in overall response rate was observed between the histological groups (P=0.24) or between patients stratified according to IPI score (P>0.05). Median overall survival, time-to-progression and response duration had not been reached after a median 19.5-month follow-up. In all, 31 patients (74%) are currently free from progression and 38 (90%) remain alive. Treatment was well tolerated. One patient (2%) experienced grade 3/4 infusion-related toxicity; 13 (31%) grade 3/4 leukopenia and 18 (43%) grade 3/4 neutropenia. Infection was observed in nine patients: eight (19%) grade 1/2 and one (2.4%) grade 3. This study demonstrates that combining rituximab with CNOP achieves high remission rates without significant additional toxicity in patients with previously untreated indolent NHL. Further follow-up will determine response duration and survival. PMID- 12750730 TI - Fertility in young women after chemotherapy with alkylating agents for Hodgkin and non-Hodgkin lymphomas. AB - INTRODUCTION: Owing to improved lymphoma cure rates, preserving fertility has received attention. MATERIAL AND METHODS: Women under 40 years at diagnosis of lymphoma, treated with >/=3 chemotherapy cycles including alkylating agents and no pelvic or total body irradiation were selected. A total of 84 women fullfiled these criteria with a median age at diagnosis at 27.4 years, the duration of first-line chemotherapy ranged from 3 to 36 months (median: 7 months) and 17 women received consolidation with high-dose therapy (HDT). After complete remission, 16 women relapsed and received second-line regimen followed by HDT for 9 months. RESULTS: With a median follow-up of 100 months, 31 women became pregnant, 34 women had primary ovarian failure (POF) and 19 women retained relative fertility. While women with preserved or relative fertility had the same clinical characteristics those with POF were older at diagnosis (30.6 versus 24.3 years), relapsed more often (34 versus 8%) and received HDT more often (54 versus 16%). After HDT (n=26), only three women (25-27 years) became pregnant. CONCLUSION: Women given alkylating agents have a high risk of POF if they are older than 30 years at diagnosis and older than 25 years at the time of HDT. For these women with a high chance of cure and who wish to be pregnant after treatment, regimens with fewer alkylating agents should be proposed or cryopreservation of embryos when possible. PMID- 12750731 TI - V(L)IgMM transgene expression in DC via a GPI-anchor using a novel retroviral vector induces an in vitro autologous T-cell proliferation restricted to MHC class I molecules. AB - INTRODUCTION: In order to improve the treatment and cure rate of multiple myeloma (MM), immunotherapy is a novel therapeutic approach. Since neoplastic plasma cells do not undergo further hypermutation, the variable region of the immunoglobulin light chain obtained from MM patients (V(L)IgMM) could serve as a tumor-specific antigen. In addition, dendritic cells (DC) have been identified as potent stimulators of an antigen-specific immune response. Here, we analyze in vitro autologous T-cell proliferation against the V(L)IgMM on presentation by retrovirally transduced dendritic cells. MATERIALS AND METHODS: Expression of the tumor antigen in DC has been achieved using a novel retroviral vector containing NH(2)(34a.a)DAF-FLAG-V(L)IgMM-DAF(37a.a)COOH transgene. After cleavage of the amino- and carboxy-terminal hydrophobic domains of DAF, the FLAG-V(L)IgMM fusion gene is attached to the membrane via a GPI-anchor molecule. Thus, transduced cells can be detected using monoclonal anti-FLAG antibodies. RESULTS: PI-PLC releases cell surface FLAG-antigen from transduced CD34(+) cells indicating that the vector directs the fusion protein to the cell surface via GPI-anchor. V(L)IgMM transgene expression in DC using our retroviral vector elicited an autologous T-cell proliferation restricted to MHC class I molecules. The proliferative response is more prominent in PMA-derived DC compared to cytokine derived DC indicating that PMA-derived DC are more potent in activating autologous T-cell proliferation. CONCLUSION: V(L)IgMM is an immunogenic peptide, which under certain conditions could provide a basis for a V(L)Ig-based immunotherapy in MM. PMID- 12750732 TI - Ten novel Diamond-Blackfan anemia mutations and three polymorphisms within the rps19 gene. AB - INTRODUCTION: A total of 25% of patients presenting with Diamond-Blackfan anemia (DBA) carry mutations in the rps19 gene, which encodes protein RPS19 of the small ribosomal subunit. The other DBA cases carry mutations in other, unknown gene(s). MATERIALS AND METHODS: We searched mutations in 48 DBA families or isolated patients based on PCR of exons of the rps19 gene and automatic sequencing. We also studied three novel intronic polymorphisms in 85 persons (most of the patients and their relatives, when the latter could be investigated). RESULTS: We identified 10 new mutations within the rps19 gene. We found no obvious correlation between the clinical expression and the nature of the mutation. Besides, we found three polymorphisms within the rps19 gene. Polymorphisms a, b and c were (i) a one-base insertion (+c at position +79) in intron 2, (ii) the g- >c substitution at position +89 also in intron 2 and (iii) the g-->a substitution at position +14 in intron 4. Inheritance studies showed that the polymorphisms were transmitted en bloc, thus defining the --- haplotypes (changes absent) and the +++ haplotypes (changes present). The percentages of each haplotype were about 50% in families and isolated persons with DBA, as well as in controls. CONCLUSION: For the 10 novel mutations found in the rps19 gene, there were no obvious genotype-phenotype correlations. The transmission of the polymorphisms was en bloc and the studies did not suggest any clinical correlates at this stage. PMID- 12750733 TI - SDF-1 enhances the expansion and maintenance of highly purified human hematopoietic progenitors. AB - The stromal-derived factor-1 (SDF-1) chemokine and its putative receptor, CXCR4, have been implicated in hematopoiesis. Here we aim to characterize the effects of cytokine-induced CXCR4 expression and SDF-1 treatment on primitive human umbilical cord blood (CB) cells in vitro. Highly purified CD34+CD38-Lin-CXCR4- blood cells were capable of forming CD34+CXCR4+ cells during short-term liquid culture, but maintained distinct erythroid and myeloid progenitor composition, similar to the parent population prior to culture. In vitro, SDF-1 enhanced the expansion and differentiation of primitive CB cells in a manner that was dependent upon both the concentration of SDF-1 and the presence of specific cytokines. In the absence of cytokine addition, cultures seeded with CD34+CD38 Lin- cells demonstrated substantial cell death; however, the addition of SDF-1 alone preferentially increased progenitor cell frequency. Our study demonstrates that induction of CXCR4 expression does not alter the differentiative potential of human blood progenitors and suggests a role for SDF-1 as a growth factor required for human hematopoiesis. PMID- 12750735 TI - Retreatment with rituximab alone induces sustained remission in a patient with follicular lymphoma with multiple extranodal sites of involvement, relapsing soon after primary treatment with fludarabine-rituximab. AB - Two recent studies have shown that retreatment of patients with relapsed indolent NHL with rituximab (RI) can be as effective as primary treatment, provided the lymphoma was initially responsive to primary RI therapy. From the available data, it remains unknown as to whether this approach is also effective for extranodal relapse. Here, we describe a 47-year-old male with stage 4B follicular lymphoma (FL), initially also involving skin, who achieved complete remission (CR) after a combination of fludarabine, cyclophosphamide and RI that lasted only 5 months. He soon relapsed with systemic disease and a number of extranodal sites including liver, lungs and bone marrow. After retreatment with RI alone (four cycles, 375 mg/m(2)), the patient achieved a second CR. Another four infusions of RI were given 6 months later as maintenance therapy. The duration of CR is already 18 months longer than the first CR. This case illustrates the fact that even in relapsed advanced FL, with multiple sites of extranodal disease, RI given as a single agent may be extremely effective in achieving an additional meaningful complete response. PMID- 12750734 TI - Developmental fate of hematopoietic stem cells: the study of individual hematopoietic clones at the level of antigen-responsive B lymphocytes. AB - We have shown previously that hematopoiesis in mice reconstituted with retrovirally marked hematopoietic stem cells (HSCs) is provided by multiple, mainly short-lived clones, as measured by retroviral insertion site analysis of individual spleen colony-forming unit (CFU-S)-derived colonies. However, the CFU S is the relatively early progenitor and the contribution of each CFU-S in the steady-state hematopoiesis is uncertain. Here, we have studied the fate of individual mature B cells, as well as CFU-S, representing the progeny of retrovirally transduced marrow-repopulating cells (MRC). B-cells-generated hybridomas and CFU-S-derived colonies were used to determine the clonal composition of hematolymphopoiesis at the single-cell level. Bone marrow (BM) cells and splenocytes (approximately 1/3-1/2 of spleen at a time) from mice reconstituted with retrovirally marked syngeneic BM cells were repeatedly collected at 3, 10, and 16 months post-transplant. The percentage of retrovirally marked CFU-S and B-cell-produced hybridomas was about 50% at 3 months and decreased to 10-15% at 10 months after reconstitution in spite of stable degree of chimerism. The clonal origin of BM-derived CFU-S and spleen-derived B-cell hybridomas was detected by Southern blot analysis. Overall, DNA obtained from 159 retrovirally marked spleen colonies, 287 hybridomas and 43 BM samples were studied. Multiple simultaneously functioning clones of MRC-derived B cells were observed. The same individual clones among hybridomas and CFU-S were identified in three out of 11 mice. Thus, hematopoiesis is generated by multiple hematopoietic clones some of which can simultaneously contribute to both mature lymphoid cells and myeloid progenitors. These data establish that the stem cell compartment functions by continuously producing progeny, which fully but transiently repopulate all lineages. PMID- 12750736 TI - Paraneoplastic pemphigus occuring after radiotherapy for relapsed non-Hodgkin's lymphoma in a patient with common variable immunodeficiency. AB - Paraneoplastic pemphigus (PNP) is a rare multisystem, autoimmune disease with prominent mucocutaneous manifestations, occurring most commonly in association with haematological malignancies. It is characterised by the presence of circulating autoantibodies against epithelial adhesion proteins. We report a 46 year-old woman with common variable immunodeficiency who developed paraneoplastic pemphigus after receiving radiotherapy for relapsed non-Hodgkin's lymphoma. Flaccid bullae covering approximately 70% of the skin, painful oropharyngeal ulceration and periocular erosions were prominent clinical features. Despite supportive treatment and attempts at disease control using high-dose corticosteroids and cyclophosphamide, the patient became increasingly debilitated, developed septic shock secondary to Pseudomonas aeruginosa septicaemia on two occasions and died of respiratory failure 6 weeks after presentation. We highlight the need to be aware of (PNP) and to perform appropriate immunological investigations. In addition, we emphasise the importance of a multidisciplinary approach to the management of such patients. PMID- 12750737 TI - Safety of rituximab in lymphoma patients with hepatitis B or hepatitis C virus infection. PMID- 12750738 TI - The dawning era of polymer therapeutics. AB - As we enter the twenty-first century, research at the interface of polymer chemistry and the biomedical sciences has given rise to the first nano-sized (5 100 nm) polymer-based pharmaceuticals, the 'polymer therapeutics'. Polymer therapeutics include rationally designed macromolecular drugs, polymer-drug and polymer-protein conjugates, polymeric micelles containing covalently bound drug, and polyplexes for DNA delivery. The successful clinical application of polymer protein conjugates, and promising clinical results arising from trials with polymer-anticancer-drug conjugates, bode well for the future design and development of the ever more sophisticated bio-nanotechnologies that are needed to realize the full potential of the post-genomic age. PMID- 12750739 TI - Drug discovery with engineered zinc-finger proteins. AB - Zinc-finger proteins (ZFPs) that recognize novel DNA sequences are the basis of a powerful technology platform with many uses in drug discovery and therapeutics. These proteins have been used as the DNA-binding domains of novel transcription factors (ZFP TFs), which are useful for validating genes as drug targets and for engineering cell lines for small-molecule screening and protein production. Recently, they have also been used as a basis for novel human therapeutics. Most of our advances in the design and application of these ZFP TFs rely on our ability to engineer ZFPs that bind short stretches of DNA (typically 9-18 base pairs) located within the promoters of target genes. Here, we summarize the methods used to design these DNA-binding domains, explain how they are incorporated into novel transcription factors (and other useful molecules) and describe some key applications in drug discovery. PMID- 12750740 TI - Hit and lead generation: beyond high-throughput screening. AB - The identification of small-molecule modulators of protein function, and the process of transforming these into high-content lead series, are key activities in modern drug discovery. The decisions taken during this process have far reaching consequences for success later in lead optimization and even more crucially in clinical development. Recently, there has been an increased focus on these activities due to escalating downstream costs resulting from high clinical failure rates. In addition, the vast emerging opportunities from efforts in functional genomics and proteomics demands a departure from the linear process of identification, evaluation and refinement activities towards a more integrated parallel process. This calls for flexible, fast and cost-effective strategies to meet the demands of producing high-content lead series with improved prospects for clinical success. PMID- 12750741 TI - Pharmacological strategies for muscular dystrophy. AB - Duchenne muscular dystrophy (DMD) is a fatal, genetic disorder whose relentless progression underscores the urgency for developing a cure. Although Duchenne initiated clinical trials roughly 150 years ago, therapies for DMD remain supportive rather than curative. A paradigm shift towards developing rational therapeutic strategies occurred with identification of the DMD gene. Gene- and cell-based therapies designed to replace the missing gene and/or dystrophin protein have achieved varying degrees of success. However, pharmacological strategies not designed to replace dystrophin per se appear promising, and can circumvent many hurdles hampering gene- and cell-based therapy. Here, we will review present pharmacological strategies, in particular those dealing with functional substitution of dystrophin by utrophin and enhancing muscle progenitor commitment by myostatin blockade, with a view toward facilitating drug discovery for DMD. PMID- 12750742 TI - Such stuff as dreams are made on: mediator-directed therapy in sepsis. AB - Sepsis, a life-threatening disorder that arises through the body's response to infection, is the leading cause of death and disability for patients in an intensive care unit. Advances in the understanding of the complex biological processes responsible for the clinical syndrome have led to the identification of many promising new therapeutic targets, including bacterial toxins, host-derived mediators, and downstream processes such as coagulation and the endocrine response. Diverse therapies directed against these targets have shown dramatic effects in animal models; however, in humans, their impact has been frustratingly modest, and only one agent--recombinant activated protein C--has achieved regulatory approval. This review summarizes the approaches that have been evaluated in clinical trials, explores the reasons for the discordance between biological promise and clinical reality, and points to approaches that may lead to greater success in the future. PMID- 12750743 TI - Is there a future for 'speculative' gene patents in Europe? AB - In the face of increasing public opposition to patents claiming property in gene sequences, is it right to deny the validity of patent applications that seem to have firm, if somewhat hypothetical, scientific basis? In this article we review the recent state of the law in Europe as it applies to gene patents in order to determine whether there is any hope for the modern heirs of Galilean innovation. PMID- 12750744 TI - NTP Carcinogenesis Bioassay of Ziram (CAS No. 137-30-4) in F344/N Rats and B6C3F1 Mice (Feed Study). AB - A NTP Carcinogenesis bioassay of ziram (89% pure, with 6.5% thiram), a fungicide and a rubber vulcanization accelerator, was conducted in F344/N rats and in B6C3F1 mice. Groups of 50 rats of each sex received diets containing 300 or 600 ppm of commercial grade ziram for 103 weeks; groups of 49 or 50 mice of each sex received diets containing 600 or 1,200 ppm ziram; and groups of 50 rats and 50 mice of each sex served as untreated controls. The average daily consumption of ziram by low-and high-dose rats, through the majority of the study, was about 11 and 22 mg/kg for males and 13 and 26 for females. The average daily consumption of ziram by low-and high-dose mice, through the majority of the study, was 122 and 196 mg/kg for males and about 131 and 248 mg/kg for females. Survival and feed consumption and mean body weights of rats of each sex were not adversely affected by ziram; rats of each sex could have tolerated higher doses. C-Cell carcinomas of the thyroid in male rats occurred with a statistically significant positive trend (P<0.01) and the incidence in the high-dose group was significantly higher (P<0.05) than that in the controls (control, 0/50, 0%; low dose, 2/49, 4%; high-dose, 7/49, 14%) and higher than that previously observed in control male rats at the same laboratory (18/584, 3%; range 0% to 8%). The combined incidence of males with either C-cell adenoma or carcinoma also showed a statistically significant (P<0.05) positive trend (control, 4/50, 8%; low-dose, 9/49, 18%; high-dose, 12/49, 24%). There were no significant histopathologic changes noted in the follicular cells. Survival of male and female mice was not adversely affected by ziram in feed; mean body weight gain by dosed male mice throughout the study and by high-dose female mice after week 80 was depressed by 15% to 20% relative to the controls. Average daily feed consumption by high-dose males and high-dose females was, respectively, 78% and 85% that of the controls. Mice probably could not have tolerated higher doses. The incidence of alveolar/bronchiolar adenomas was significantly (P<0.05) increased in female mice (control, 2/50, 4%; low-dose, 5/49, 10%; high-dose, 10/50, 20%). The combined incidence of alveolar/bronchiolar adenomas or carcinomas in female mice showed a statistically significant (P<0.05) positive trend. The incidence in the high-dose group was significantly (P<0.05) higher than that in the controls (control, 4/50, 8%; low-dose, 6/49, 12%; high-dose, 11/50, 22%). Pulmonary adenomatous hyperplasia consistent with chronic Sendai virus infection (confirmed by serologic analysis performed on untreated animals from the same animal shipment and present in the same room) was observed in control and dosed male mice (control, 15/49, 31%; low-dose, 19/50, 38%; high-dose, 16/49, 33%) as well as in control and dosed female mice (control, 18/50, 36%; low-dose, 27/49, 55%; high dose, 26/50, 52%). Six of the 26 high-dose females without pulmonary adenomatous hyperplasia had pulmonary tumors, whereas 4 of the 24 high-dose females without pulmonary adenomatous hyperplasia also had pulmonary tumors. Only 1 of 27 low dose females with adenomatous hyperplasia had a pulmonary tumor. There was a significant decrease in the incidence of mammary fibroadenomas in high-dose female rats (control, 16/50, 32%; low-dose, 17/50, 34%; high-dose, 8/50, 16%). Significant dose-related decreased incidences of liver carcinomas in male mice (control, 13/49, 27%; low-dose, 8/50, 16%; high-dose, 1/49, 2%) and of liver adenomas in female mice (control, 7/50, 14%; low-dose, 2/50, 4%; high-dose, 0/50, 0%) were observed. Under the conditions of these studies, ziram was carcinogenic for male F344/N rats, causing increased incidences of C-cell carcinomas of the thyroid gland. Ziram was not carcinogenic for either female F344/N rats or for male B6C3F1 mice. Increased incidences of alveolar/bronchiolar adenomas and of combined alveolar/bronchiolar adenomas or carcinomas occurred in female B6C3F1 mice. However, the interpretation of this increase in lung tumors is complicat either female F344/N rats or for male B6C3F1 mice. Increased incidences of alveolar/bronchiolar adenomas and of combined alveolar/bronchiolar adenomas or carcinomas occurred in female B6C3F1 mice. However, the interpretation of this increase in lung tumors is complicated by an intercurrent Sendai virus infection. Levels of Evidence of Carcinogenicity: Male Rats: Positive Female Rats: Negative Male Mice: Negative Female Mice: Equivocal Synonym: zinc dimethyldithiocarbamate PMID- 12750745 TI - NTP Carcinogenesis Studies of 4,4'-Methylenedianiline Dihydrochloride (CAS No. 13552-44-8) in F344/N Rats and B6C3F1 Mice (Drinking Water Studies). AB - 4,4'-Methylenedianiline is used primarily as a chemical intermediate in the closed system production of isocyanates and polyisocyanates. These chemicals are used extensively in the manufacture of rigid polyurethane foams for thermal insulation and in the production of semiflexible polyurethane foams for automobile safety cushioning. The saturated isocyante of 4,4'-methylenedianiline [4,4'-methylene-bis(cyclohexylisocyanate)] is an intermediate in the production of light-stable, high-performance polyurethane coatings. 4,4'-Methylenedianiline is also a curing agent for epoxy resins and urethane elastomers, a dye intermediate, and a corrosion inhibitor. NTP Carcinogenesis studies of 4,4' methylenedianiline dihydrochloride (98.6% pure) were conducted by administering this chemical in the drinking water of F344/N rats and B6C3F1 mice. Groups of 50 rats and 50 mice of each sex received drinking water containing 150 or 300 ppm 4,4'-methylenedianiline dihydrochloride (dosage expressed as the free base) for 103 weeks. Groups of 50 rats and 50 mice of each sex, given drinking water adjusted with 0.1N HCl to the pH (3.7) of the 300-ppm formulation, served as controls. Survival was comparable among groups except for male mice receiving the high dose of 4,4'-methylenedianiline dihydrochloride; survival in that group was lower (P=0.006) than that in controls. Mean body weight was reduced in high dose female rats and in high dose male and female mice. Water consumption was reduced in a dose-related manner in both sexes of rats. No compound-related clinical effects were observed. Compound-related nonneoplastic lesions of the thyroid in female rats included follicular cysts and hyperplasia. The incidence of thyroid follicular cell hyperplasia was elevated in high dose male and female mice. The incidences of thyroid neoplasms in the high dose groups were elevated compared with those of the control groups for both sexes of both species. Thyroid follicular cell carcinoma was increased in male rats (controls, 0/49; low dose, 0/47; high dose, 7/48, 15%: P97% pure) was conducted by administering diets containing 25,000 or 50,000 ppm L-ascorbic acid to groups of 50 F344/N rats and 50 B6C3F1 mice of each sex for 103 weeks. Controls consisted of 50 untreated rats and untreated mice of each sex. Fifty-thousand ppm is the highest dose recommended for chronic studies. Survival of dosed and control female rats and of dosed and control female mice were comparable. Survival of high-dose male rats was slightly greater than that of the controls (P=0.087). Survival of high-dose male mice was significantly greater (P=0.009) than that of the controls. Throughout most of the study, mean body weights of dosed female rats and dosed female mice were lower than those of the controls. Final body weights were comparable among groups, except for the high-dose female rats (<13%); marginal differences (<8%) were observed for low-dose female rats and for dosed female mice (8%-11%). Food consumption was equivalent among groups. Most observational differences were confined to the female rat. The incidence of low dose female rats with undifferentiated (mononuclear-cell) leukemias (control, 6/50, 12%; low-dose, 17/50, 34%; high-dose, 12/50, 24%) was significantly higher (P<0.02) than that in controls. These tumors were not considered to be related to administration of L-ascorbic acid because they did not occur in the female high dose group at incidences significantly greater (P>0.07) than those in the controls, the trend test was not significant (P>/=0.07), and no increases were observed for male rats. Under the conditions of this bioassay, L-ascorbic acid was not carcinogenic for male and female F344/N rats or male and female B6C3F1 mice. Levels of Evidence of Carcinogenicity: Male Rats: Negative Female Rats: Negative Male Mice: Negative Female Mice: Negative Synonym: vitamin C PMID- 12750747 TI - NTP Lifetime Carcinogenesis Studies of Chrysotile Asbestos (CAS No. 12001-29-5) in Syrian Golden Hamsters (Feed Studies). AB - NTP Carcinogenesis studies of short range (SR), intermediate range (IR), or intermediate range chrysotile asbestos in combination with the intestinal carcinogen 1,2-dimethylhydrazine dihydrochloride (DMH) were conducted with male and female Syrian golden hamsters. Both forms of chrysotile asbestos were administered at the concentration of 1% in pelleted diet for the entire lifetime of the hamsters starting with mothers of the test animals. Group sizes varied from 125 to 253. Starting at 6 weeks of age, male and female hamsters in the intermediate range chrysotile/DMH study were given oral doses of DMH (4 mg/kg) every other week for a total of 5 doses. There was no adverse effect on body weight gain or survival by either form of asbestos or by asbestos in combination with DMH. A significant increase (P< 0.05) in adrenal cortical adenomas was observed in male hamsters exposed to SR and IR chrysotile asbestos and in females treated with IR chrysotile asbestos when compared to the pooled control groups (males: pooled controls, 25/466, 5%; SR chrysotile, 26/299, 11%; IR chrysotile, 24/244, 10%; females: pooled controls, 15/468, 3%; IR chrysotile, 18/234, 8%). However, statistical significance was lost when these dosed groups were compared with concurrent control groups (males: SR control, 7/115, 6%; IR control, 7/115, 6%; females: SR control, 4/112, 4%; IR control, 6/118, 5%). The results of the combination study (IR chrysotile plus DMH) did not yield a significant increase in tumors above the background level observed in the DMH group alone or in the untreated control group. The DMH failed to yield a background level of intestinal tumors high enough to provide a valid test of the cocarcinogenic potential of chrysotile asbestos. For this reason, the cocarcinogenic potential of orally administered asbestos should be considered untested. Under the conditions of these studies, neither short range chrysotile nor intermediate range chrysotile asbestos was carcinogenic when ingested at 1% levels in the diet by male and female Syrian golden hamsters. While there were increases in the rates of adrenal cortical adenomas in male and female hamsters exposed to intermediate range chrysotile asbestos compared to the pooled groups, these incidence rates were not different when compared with the concurrent control groups. Additionally, the biologic importance of adrenal tumors in the absence of target organ (gastrointestinal tract) neoplasia is questionable. The coNTP Carcinogenesis studies using IR chrysotile asbestos and 1,2-dimethylhydrazine dihydrochloride were considered inadequate because there was no increase in intestinal neoplasia in the DMH group. Levels of Evidence of Carcinogenicity: Intermediate Range: Male Hamsters: Negative Female Hamsters: Negative Intermediate Range + dimethyl hydrazine: Male Hamsters: Inadequate Female Hamsters: Inadequate Short Range: Male Hamsters: Negative Female Hamsters: Negative. PMID- 12750748 TI - NTP Carcinogenesis Bioassay of Melamine (CAS No. 108-78-1) in F344/N Rats and B6C3F1 Mice (Feed Study). AB - A NTP Carcinogenesis bioassay of melamine (>95% pure), a chemical intermediate in the manufacture of amino resins and plastics, was conducted by feeding diets containing 2,250 or 4,500 ppm melamine to groups of 50 male F344/N rats and 50 B6C3F1 mice of each sex for 103 weeks. Groups of 49 male rats, 50 female rats, 49 male mice, and 50 female mice served as controls. Mean body weights of dosed rats of each sex were lower than those of the controls after week 20. Survival of high dose male rats was significantly lower (P=1 mg/l), and the detection of mutations involved in resistance is used in many laboratories. We analyzed 36 clarithromycin-resistant strains isolated from children and 30 from adults. In vitro susceptibility to clarithromycin was determined by an agar dilution method. DNA from the isolates was extracted using the method published by Ge and Taylor. A2142G and A2143G mutations were identified by PCR-RFLP. A 1.4 Kpb of the 23S rRNA gene was amplified and digested using MboII or BsaI restriction enzymes to detect mutations. The prevalence of the A-G transition mutation at position 2143 was higher in the children (80.55%) than in the adult patients (46.66%) (p<0.05); however, the prevalence of the mutation at position 2142 was higher in adults than in children (36.66% vs. 5.55%; p<0.05). In children, a higher MIC (2-64 mg/l) was observed when the A-G mutation was detected at position 2143. However, in adult patients higher MICs were observed when the A-G mutation was detected at position 2142. PMID- 12750758 TI - [Susceptibility of Streptococcus pyogenes isolates from pharyngeal exudates in Cordoba (Spain)]. AB - Streptococcus pyogenes is an important human pathogen. Betalactams are still the drug of choice for the treatment of infections caused by this microorganism. In recent years an increase in the use of macrolides for initial treatment in respiratory infections has been observed; consequently, the number of macrolide resistant isolates has also increased. We investigated the susceptibility of S. pyogenes to penicillin, erythromycin, clarithromycin and clindamycin in Cordoba during 2000, 2001 and the first 6 months of 2002. We obtained 100 isolates of S. pyogenes from 1232 pharyngeal exudates, all of which were susceptible to penicillin and 39 of which were resistant to erythromycin and clarithromycin. Twenty-six of these 39 isolates were susceptible to clindamycin. PMID- 12750759 TI - [Susceptibility of Streptococcus pyogenes to macrolides and quinolones in Guadalajara, Spain]. AB - We carried out a retrospective study of the susceptibility of 104 Streptococcus pyogenes strains, which were isolated in 2000 and 2001 from clinical samples of different origins, to penicillin, erythromycin, clindamycin, ofloxacin and levofloxacin. The susceptibility testing was performed using the agar difusion method according to the guidelines of the NCCLS. All of the isolates showed susceptibility to penicillin and clindamycin. However, we detected 11 strains that were resistant to erythromycin (10.6%) and 4 strains resistant to ofloxacin (3.8%). We studied the resistance phenotypes of macrolides and lincosamides using erythromycin and clindamycin discs. Nine of the eleven strains that were resistant to erythromycin showed an M phenotype, while the remaining two showed inducible resistance to clindamycin, thus suggesting an MLS(B) inducible phenotype. No strains with constitutive resistance to erythromycin or clindamycin (MLS(B) constitutive phenotype) were identified. While penicillin is still uniformly active against S. pyogenes, in Guadalajara, there are 10.6% strains that are resistant to 14- and 15-atoms macrolides. PMID- 12750760 TI - [Antiretroviral therapy from 1996-2000]. AB - A retrospective study was carried out from September 1996 to June 2000 and examined the antiretroviral therapies prescribed to HIV patients in whom the viral load had been determined. Triple therapy was the most frequently prescribed during the study and the annual percentage increased to 60% in the last semesters analyzed. Large variability in antiretroviral combinations was observed and depended on the clinical center and unit. The number of individuals who began viral-load monitoring before being treated with antiretroviral drugs showed a progressive increase. PMID- 12750761 TI - 5-HT and the treatment of irritable bowel syndrome: A clinical perspective. AB - The gut contains large amounts of serotonin (5-HT) and this neuro-transmitter is now known to be intricately involved in the control of gastrointestinal physiological function via a number of receptor subtypes. The 5-HT(3) and 5-HT(4) receptors are currently the focus of much attention with respect to the therapy of irritable bowel syndrome (IBS) by the use of either agonists or antagonists. This article concentrates on the development of alosetron, cilansetron and tegaserod, highlighting the change in thinking that has to accompany the use of these new drugs, especially with regard to targeting subgroups and avoiding inappropriate prescription. It also raises the philosophical question of how safe drugs should have to be in this therapeutic area. (c) 2001 Prous Science. All rights reserved. PMID- 12750762 TI - Nonpeptide endothelin antagonists in clinical development. AB - Endothelins (ET-1, ET-2 and ET-3) are 21-amino-acid peptides with two disulfide bonds that belong to the sarafotoxin family. ET-1, ET-2 and ET-3 are produced endogenously from preproendothelin to give big endothelins, which are cleaved by endothelin-converting enzyme (ECE) to yield the active protein. Endothelin has been shown to play important physiological and pathological roles by interacting with its G-protein-coupled receptors. There are two cloned ET receptors: the ET(A) receptor, which is selective for ET-1, and the ET(B) receptor, which binds ET-1, ET-2 and ET-3 with similar affinities. Since the discovery of endothelin, and especially since the availability of peptide ET antagonists such as BQ-123 and BQ-788, and nonpeptide compounds such as bosentan, considerable effort has been spent on better understanding the role of endothelin and its receptor antagonists. As a result, endothelin has been implicated in a variety of serious diseases, such as congestive heart failure, hypertension, pulmonary hypertension and prostate cancer. Research in pharmaceutical and biotechnology laboratories has generated many endothelin antagonists with either sulfonamide or triaryl carboxylic acid scaffolds, and a number of ET(A)-selective or nonselective ET(A)/ET(B) endothelin antagonists have entered clinical trials. This article will review the small-molecule ET(A)-selective and nonselective ET(A)/ET(B) antagonists that are under clinical evaluation, and highlight a member of this group of compounds, sitaxsentan. A summary of the medicinal chemistry that led to the identification of sitaxsentan will be presented, followed by selected animal and human clinical trial data. (c) 2001 Prous Science. All rights reserved. PMID- 12750763 TI - New therapeutic approaches to the treatment of nasal allergy: Antiinflammatory effects of H(1) receptor antagonists. AB - Histamine is recognized as an important mediator of allergic rhinitis. In addition to the role it plays in the immediate-phase reaction of nasal allergy, histamine may play an important role in the late-phase reaction and protracted allergic inflammation. Histamine induces the synthesis or secretion of proinflammatory cytokines and the expression of adhesion molecules. Besides their traditional effects via H(1) receptor antagonism, antihistamines have been reported to possess multiple antiinflammatory effects. Continuous use of antihistamines may reduce the level of minimal persistent inflammation of nasal mucosa in allergy. Treatment of allergic rhinitis using antihistamines could contribute to the improvement in coexisting allergic inflammation of the lower airways. (c) 2001 Prous Science. All rights reserved. PMID- 12750764 TI - Brain natriuretic peptide: Disease marker or more in cardiovascular medicine? AB - Brain natriuretic peptide (BNP) is predominantly a cardiac ventricular hormone that promotes natriuresis and diuresis, inhibits the renin-angiotensin aldosterone axis, and is a vasodilator. Plasma BNP levels are raised in essential hypertension, and more so in left ventricular (LV) hypertrophy and heart failure. Plasma BNP levels are also elevated in ischemic heart disease. Attempts have been made to use plasma BNP levels as a marker of LV dysfunction, but these have shown that plasma BNP levels are probably not sensitive enough to replace echocardiography in the diagnosis of LV dysfunction. Pericardial BNP or N-BNP may be more suitable markers of LV dysfunction. Plasma BNP levels are also elevated in right ventricular dysfunction, pregnancy-induced hypertension, aortic stenosis, age, subarachnoid hemorrhage, cardiac allograft rejection and cavopulmonary connection, and BNP may have an important pathophysiological role in some or all of these conditions. Clinical trials have demonstrated the natriuretic, diuretic and vasodilator effects, as well as inhibitory effects on renin and aldosterone of infused synthetic human BNP (nesiritide) in healthy humans. BNP infusion improves LV function in patients with congestive heart failure via a vasodilating and a prominent natriuretic effect. BNP infusion is useful for the treatment of decompensated congestive heart failure requiring hospitalization. The clinical potential of BNP is limited as it is a peptide and requires infusion. Drugs that modify the effects of BNP are furthering our understanding of the pathophysiological role and clinical potential of BNP. Increasing the effects of BNP may be a useful therapeutic approach in heart failure involving LV dysfunction. The levels of plasma BNP are increased by beta blockers, cardiac glycosides and vasopeptidase inhibitors, and this may contribute to the usefulness of these agents in heart failure. (c) 2001 Prous Science. All rights reserved. PMID- 12750765 TI - The role of chemokines and adhesion molecules in the pathogenesis of rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is characterized by marked infiltration of the synovium by T-cells. The mediator of joint inflammation has been shown to be both cellular and soluble and the concept of cross-talk between adhesion molecules and cytokines, and its relevance to inflammation, is emerging. The expression and function of adhesion molecules are tightly regulated via intracellular signaling induced by cytokine or chemokine stimulation, a process which is designated "inside-to-out signaling". Such regulation is particularly important in inflammatory processes in which T-cells migrate from the circulation into the tissue. Adhesion molecules not only function as glue, but also transduce extracellular information into cytoplasmic organelles via the "outside-to-in signal", resulting in cell activation and cytokine production. For instance, the abundant intracellular adhesion molecule ICAM-1 and CD44 on RA synoviocytes not only potentially facilitate the interaction with T-cells or extracellular matrix, but also induce cytokine gene transcription in synovial cells. Thus, the two directional cross-talk among adhesion molecules and cytokines appears to be significant for the initiation and prolongation of inflammatory processes through T-cell migration into RA synovial tissues and activation of both T-cells and synovial cells in this region. The concept proposed would greatly help in clarifying the pathological processes in rheumatoid synovitis, as well as in discovering new pharmacological approaches to more specifically control synovial inflammation. (c) 2001 Prous Science. All rights reserved. PMID- 12750766 TI - Recent advances in cancer research. Highlights of the 92nd annual meeting of the American Association for Cancer Research; held March 24-28, 2001, in New Orleans, LA, U.S.A. AB - The 92nd Annual Meeting of the American Association for Cancer Research (AACR) was held on March 24-28, 2001, in New Orleans, Louisiana, U.S.A., to discuss the latest advances in basic, translational and clinical cancer research. More than 12,000 scientists representing academia, industry and government attended this international meeting. Over 5,300 proffered papers were presented in a variety of formats. This meeting report highlights a lecture on the antileukemia drug STI 571 (imatinib) and minisymposia that covered the topics of tumor immunity, immunotherapy, pharmacogenetics and novel targets for drug discovery. (c) 2001 Prous Science. All rights reserved. PMID- 12750767 TI - IDDM2/insulin VNTR modifies risk conferred by IDDM1/HLA for development of Type 1 diabetes and associated autoimmunity. AB - AIM/HYPOTHESIS: Type 1 diabetes (T1D) is an autoimmune disease with multiple susceptibility genes. The aim of this study was to determine whether combining IDDM1/HLA and IDDM2/ insulin( INS) 5' variable number of tandem repeat locus (VNTR) genotypes improves T1D risk assessment. METHODS: Patients with T1D (n=488), control subjects (n=846), and offspring of parents with T1D (n=1122) were IDDM1 and IDDM2 genotyped. Offspring were followed for islet autoantibodies and T1D from birth until the age of 2 to 12 years. RESULTS: Compared to the I/I INS VNTR genotype, the I/III and III/III genotypes reduced T1D risk conferred by IDDM1/HLA in all HLA genotype categories of the case-control cohort by 1.6-fold to three-fold. The highest T1D risk was associated with INS VNTR class I/I plus HLA DR3/DR4-DQ8 (20.4% in patients, 0.6% in control subjects) or HLA DR4-DQ8/DR4 DQ8 (6.3% in patients, 0.2% in control subjects). In the offspring, HLA DR3/DR4 DQ8 and DR4-DQ8/DR4-DQ8 conferred increased risk for early development of islet autoantibodies (14.6% and 12.9% by age 2 years). Offspring with these high risk IDDM1 genotypes plus the INS VNTR class I/I genotype (n=71; 6.3%) had the highest risk of developing islet autoantibodies (21.8% by age 2 years vs 8.9% in offspring with high risk IDDM1 plus INS VNTR class I/III or III/III genotypes, p<0.05) and T1D (8.5% by age 6 years vs 4.3%). Offspring who developed autoantibodies to multiple antigens had increased frequencies of both high risk IDDM1 and IDDM2 genotypes (p<0.0001), whereas offspring who developed autoantibodies to GAD only had increased frequencies of high risk IDDM1 and protective IDDM2 genotypes, suggesting that IDDM2 influences the autoimmune target specificity. CONCLUSION/INTERPRETATION: Combining IDDM1 and IDDM2 genotyping identifies a minority of children with an increased T1D risk. PMID- 12750768 TI - Association between macrophage activation and function of micro-encapsulated rat islets. AB - AIMS/HYPOTHESIS: Survival of microencapsulated islet grafts is limited, even when inflammatory reactions against the capsules are restricted to a small portion of less than 10%. METHODS: This study investigates both in vivo in rat recipients and in vitro whether cellular overgrowth on this minority of the capsules contributes to limitations in the functional survival of the 90% of the encapsulated islets which remain free of any cellular overgrowth. RESULTS: In successful rat recipients of an allogenic microencapsulated islet graft we found that the vast majority of cells in the capsular overgrowth were activated ED-1 and ED-2 positive macrophages which were found in numbers of approximately 1500 per capsule. Co-culture of encapsulated islets with 1500 (nr8383) rat-macrophages per capsule showed that the activation of macrophages was caused by islet-derived bioactive factors since TNF-alpha and IL-1beta secretion by macrophages was induced by islet-containing capsules and not by empty capsules. This activation of macrophages was associated with a decrease in function of the encapsulated islets as evidenced by a quantitatively reduced (35%) insulin response in static incubation and a slower response in perifusion. CONCLUSION/INTERPRETATION: Present research aims to design strategies for the temporary inhibition of macrophage activation since macrophages are predominantly present in the first two months after implantation. These strategies will serve as a pertinent basis for future clinical application of microencapsulated islets. PMID- 12750769 TI - Gender difference in all-cause and cardiovascular mortality related to hyperglycaemia and newly-diagnosed diabetes. AB - AIM/HYPOTHESIS: Diabetic women generally have a greater relative risk of cardiovascular diseases than diabetic men in comparison with non-diabetic women and men. Reasons for this excess risk in diabetic women is still unclear. The aim of this study is to evaluate whether the association between different degrees of hyperglycaemia and the risk of all-cause and cardiovascular mortality is different in women and men. METHODS: We analysed baseline glucose concentrations from 14 prospective European cohorts including 8172 men and 9407 women aged 30 to 89 years without history of diabetes, with a median follow-up of 8.3 years. Hazards ratios for all-cause and cardiovascular mortality were estimated adjusting for other risk factors. RESULTS: The mortality rates for all-cause and cardiovascular diseases were higher in men than in women in normoglycaemia, impaired glucose regulation and newly-diagnosed diabetes; the largest sex differential for cardiovascular mortality was in normoglycaemic people. The hazards ratios for all-cause and cardiovascular mortality were higher in newly diagnosed diabetic women than men compared with normoglycaemic women and men, respectively; however, this sex difference was only significant for cardiovascular mortality. For smokers and for subjects with hypertension, hypercholesterolaemia or who where overweight, the hazards ratios for cardiovascular mortality in diabetic patients compared with normoglycaemic people were also higher in women than in men. CONCLUSIONS/INTERPRETATION: Newly diagnosed diabetic women showed higher relative risks for death from cardiovascular disease than diabetic men. Thus a more aggressive control of hyperglycaemia as well as of other cardiovascular risk factors might be appropriate in women with asymptomatic hyperglycaemia. PMID- 12750771 TI - Impact of the Coffea canephora gene introgression on beverage quality of C. arabica. AB - Lines of Coffea arabica derived from the Timor Hybrid (hybrid between C. arabica and C. canephora) are resistant to coffee leaf rust (Hemileia vastatrix) and to the nematode Meloidogyne exigua. The introgression of C. canephora resistance genes is suspected of causing a drop in beverage quality. Coffee samples from pure lines, compared in a Trial 1, and from F1 hybrids and parental lines from a half-diallel trial in a Trial 2, were studied for beverage quality, chemical composition and amount of introgressed genetic material. Chemical analyses (caffeine, chlorogenic acids, fat, trigonelline, sucrose) were carried out with near-infrared spectrometry by reflectance of green coffee. The number of amplified fragment length polymorphic (AFLP) markers introgressed from the Timor Hybrid varied from 1 to 37 for the lines studied. There were significant differences between lines for all of the biochemical compounds analysed and for the acidity and the overall standard of the beverage. Two lines (T17927, T17924) were significantly poorer than the controls for sucrose and beverage acidity. T17924 also had more chlorogenic acids and was poorer for the overall standard. However, two highly introgressed lines, T17934 and T17931 (25 and 30 AFLP markers, respectively), did not differ from the non-introgressed controls. There were no correlations between the number of AFLP markers and the chemical contents or beverage attributes. Significant correlations were found between the performance of the parents and their general combining ability for beverage quality. It was concluded that it should be possible to find lines with both the desired resistance genes and good beverage quality. Selection can avoid accompanying the introgression of resistance genes with a drop in beverage quality. PMID- 12750770 TI - IRS1 degradation and increased serine phosphorylation cannot predict the degree of metabolic insulin resistance induced by oxidative stress. AB - AIM/HYPOTHESIS: Oxidative stress was shown to selectively induce impaired metabolic response to insulin, raising the possible involvement of alterations in Insulin-Receptor-Substrate (IRS) proteins. This study was conducted to assess whether oxidative stress induced IRS protein degradation and enhanced serine phosphorylation, and to assess their functional importance. METHODS: 3T3-L1 adipocytes and rat hepatoma cells (FAO) were exposed to micro-molar H(2)O(2) by adding glucose oxidase to the culture medium, and IRS1 content, its serine phosphorylation and downstream metabolic insulin effects were measured. RESULTS: Cells exposed to oxidative stress exhibited decreased IRS1 (but not IRS2) content, and increased serine phosphorylation of both proteins. Total protein ubiquitination was increased in oxidized cells, but not in cells exposed to prolonged insulin treatment. Yet, lactacystin and MG132, two unrelated proteasome inhibitors, prevented IRS1 degradation induced by prolonged insulin but not by oxidative stress. The PI 3-kinase inhibitor LY294002 and the mTOR inhibitor rapamycin, but not the MEK1 inhibitor PD98059, could prevent IRS1 changes in oxidized cells. Rapamycin, which protected against IRS1 degradation and serine phosphorylation was not associated with improved response to acute insulin stimulation. Moreover, the antioxidant alpha lipoic acid, while protecting against oxidative stress-induced insulin resistance in 3T3-L1 adipocytes, could not prevent IRS1 degradation and serine phosphorylation. CONCLUSION/INTERPRETATION: Oxidative stress induces serine phosphorylation of IRS1 and increases its degradation by a proteasome-independent pathway; yet, these changes do not correlate with the induction of impaired metabolic response to insulin. PMID- 12750772 TI - Characterization of a mosaic minisatellite locus in the mitochondrial DNA of Norway spruce [Picea abies (L.) Karst.]. AB - A mosaic minisatellite region has been identified in the mitochondrial genome of Norway spruce (Picea abies). The array was composed of three tandem repeats PaTR1 (32 bp), PaTR2a (26 bp) and PaTR2b (26 bp). PaTR2a and PaTR2b differed by one base substitution. The analysis of 92 trees covering the whole natural distribution area of the species allowed detection of 11 length variants ranging from 131 bp to 447 bp. This high intra-specific polymorphism relies on variation in the number of the tandem repeats. Population genetic parameters estimated among 14 populations suggested high population differentiation (Gst=0.749). The phylogenetic analysis of the 11 sequenced length variants has been performed using a parsimony approach. The topology of the tree showed a good association of groups with geographical origin and a low level of size homoplasy. The phylogenetic reconstruction also suggests that this minisatellite locus has mainly evolved by an increase in the repeat copy number. PMID- 12750773 TI - Genetic evaluation of alternative silvicultural systems in coastal montane forests: western hemlock and amabilis fir. AB - Genetic diversity and mating system were quantified for shelterwood, patch cut and green tree-retention silvicultural systems, and compared to adjacent old growth. This is a component of a larger study conducted in montane old-growth forests of coastal British Columbia to evaluate the feasibility and ecological consequences of alternative silvicultural systems. The experiment includes replicated treatments representing a range of overstory removal adjacent to old growth and clearcut areas. Based on 22 electrophoretically assayed loci, the effects of silvicultural systems on genetic parameters of amabilis fir (Abies amabilis and western hemlock (Tsuga heterophylla were assessed by comparing an average number of alleles per locus, the percent polymorphic loci, and observed and expected heterozygosity between parental populations and naturally regenerated progeny as well as among treatments. Genetic variation in natural regeneration was greater than in parental populations, especially for low frequency alleles. Silvicultural treatments caused no significant differences in amabilis fir genetic-diversity parameters, while the shelterwood system resulted in lower observed and expected heterozygosity in western hemlock. Nei's genetic distance revealed that all parental populations were extremely similar. The two species had contrasting mating system dynamics with amabilis fir producing higher levels of correlated paternity and inbreeding with wider variation among individual tree outcrossing-rate estimates. Western hemlock had significant levels of correlated paternity only for the green tree and shelterwood treatments demonstrating family structuring inversely related to stand density. Inbreeding in western hemlock was significant but lower than that observed for amabilis fir with a J-shaped distribution for individual tree multilocus outcrossing-rate estimates. The pollination and dispersal mechanisms of the two species represent the most-likely factors causing these differences. Artificial regeneration may be utilized to augment the genetic resources of natural ingress. PMID- 12750774 TI - Bankuti 1201--an old Hungarian wheat variety with special storage protein composition. AB - Bankuti 1201, an old Hungarian wheat variety with special quality traits, was analysed to determine the relationships between its storage protein composition and superior quality-attributes for breadmaking. Based on the storage protein composition, the variety appears to have the nature of a population, containing several genotypes with different gluten protein alleles. Using molecular markers, a new mutant x-type HMW glutenin allele was identified, containing an extra cysteine residue and showing a moderate, positive-effect on gluten properties. In lines possessing subunits Bx7+By8 the overexpression of the Bx-type subunit could be detected, resulting in a higher unextractable polymeric protein (UPP) content and increased dough strength. It was found that the presence or absence of subunit Bx7 has an equilibrating effect on the dough extensibility, which is generally characteristic of the Bankuti 1201 population. The complex good bread making quality of the variety, which has strong but highly extensible dough, is probably due to the balance between lines which express subunit Bx7 and those which do not. PMID- 12750775 TI - Gene flow estimation with microsatellites in a Malagasy seed orchard of Eucalyptus grandis. AB - Eucalyptus grandis has a mixed-mating reproductive system. Malagasy Eucalyptus seed orchards were established 15 years ago with two aims both based on panmixia: open-pollinated seed production and genetic improvement. The panmixia hypothesis has never been confirmed in the seed orchard. From a seedling seed-orchard stand comprising 349 trees and using data obtained with six selected microsatellite markers, paternity analysis was performed for 724 offspring collected on 30 adult trees. Paternity assignment, based on exclusion procedures and likelihood-ratio method, was achieved with high accuracy; the exclusion probability value was 0.997. The outcrossing rate was very high (96.7%). More than 50% of potential male trees (199 out of 349) in the seed orchard contributed to pollination for 440 offspring of 30 progenies (8.6% of the basic population). The pollination rate from outside the seed orchard was high (39.2%), but might be due to the small size of this seed orchard. This study showed that "panmixia-like pollination" can be assumed. PMID- 12750776 TI - Distinction between cultivated and wild chicory gene pools using AFLP markers. AB - The cultivation area of industrial chicory, Cichorium intybus L. cv Sativum, coincides with the natural distribution area of its wild relative, C. intybus L., which could lead to gene flow between wild and cultivated types. The genetic diversity within and between the two types has therefore been studied using AFLP genotyping of samples from 12 wild populations collected in Belgium and ten commercial varieties. The genotyping of 233 individuals allowed the identification of 254 AFLP markers. Similar levels of genetic diversity were observed within wild populations and cultivated varieties, suggesting the absence of any strong bottleneck in the history of the cultivated types. The phylogenetic analysis pointed to a monophyletic origin of cultivated varieties as compared to the local wild populations studied, hence the two types of chicory form two separate gene pools. The genotyping of some individuals sampled in ruderal sites clearly showed that they belong to the cultivated gene pool, which suggests the existence of feral or weedy types. The low differentiation observed among wild populations indicates that gene flow might be important in this species. PMID- 12750777 TI - Cotyledon storage proteins as markers of the genetic diversity in Castanea sativa Miller. AB - This study has been to analyse the useful nut globulin proteins as a marker of the genetic diversity in Castanea sativa. The evaluated populations were highly polymorphic for the globulins, being detected up to 35 polymorphic bands with a wide distribution among all the evaluated populations. Taken together for populations from all the chestnut regions, about 39.3% of total allelic variation was distributed among the populations. The estimates of genetic similarity between populations were clearly associated with the collecting site. This method of analysis of the nut storage proteins (globulins) could be a useful tool for the evaluation of genetic diversity in this and other species of the Fagaceae. PMID- 12750778 TI - Microsatellite variability in peach [ Prunus persica (L.) Batsch]: cultivar identification, marker mutation, pedigree inferences and population structure. AB - A collection of 212 peach and nectarine cultivars covering a wide variation of the species were studied with 16 polymorphic single-locus microsatellite, or simple-sequence repeat (SSR), markers. The average number of alleles per locus was 7.3, 35% of the cultivar x locus combinations analyzed were heterozygous and 87% of the cultivars studied could be individually identified. Most of the groups where two or more cultivars had the same SSR fingerprint included known peach mutants or possible synonymies. Pedigree information was tested with the SSR data. Five unexpected genotypes, due to a mutation at five SSR loci were found when comparing the SSR fingerprint of 14 known mutant cultivars and putative synonymous cultivars. The pedigree data were not consistent with the observed data in 11 out of 38 cases that could be analyzed. The group of non-melting fruit flesh cultivars, generally used by the canning industry, was more variable and genetically distant than the rest of the cultivars tested. Based on their level of homozygosity it was possible to separate those cultivars that were obtained by modern breeding technologies from those that were selected from traditional orchards after generations of seed propagation. The former had a distribution of genotypic frequencies close to a random mating model while the latter had a higher level of homozygosity. The implications of these data for the use of SSR fingerprints in breeder's rights protection and peach breeding are discussed. PMID- 12750779 TI - Cross-species amplification of mitochondrial DNA sequence-tagged-site markers in conifers: the nature of polymorphism and variation within and among species in Picea. AB - Primers previously developed to amplify specific non-coding regions of the mitochondrial genome in Angiosperms, and new primers for additional non-coding mtDNA regions, were tested for their ability to direct DNA amplification in 12 conifer taxa and to detect sequence-tagged-site (STS) polymorphisms within and among eight species in Picea. Out of 12 primer pairs, nine were successful at amplifying mtDNA in most of the taxa surveyed. In conifers, indels and substitutions were observed for several loci, allowing them to distinguish between families, genera and, in some cases, between species within genera. In Picea, interspecific polymorphism was detected for four loci, while intraspecific variation was observed for three of the mtDNA regions studied. One of these (SSU rRNA V1 region) exhibited indel polymorphisms, and the two others ( nad1 intron b/c and nad5 intron1) revealed restriction differences after digestion with Sau3AI (PCR-RFLP). A fourth locus, the nad4L- orf25 intergenic region, showed a multibanding pattern for most of the spruce species, suggesting a possible gene duplication. Maternal inheritance, expected for mtDNA in conifers, was observed for all polymorphic markers except the intergenic region nad4L- orf25. Pooling of the variation observed with the remaining three markers resulted in two to six different mtDNA haplotypes within the different species of Picea. Evidence for intra-genomic recombination was observed in at least two taxa. Thus, these mitotypes are likely to be more informative than single-locus haplotypes. They should be particularly useful for the study of biogeography and the dynamics of hybrid zones. PMID- 12750781 TI - Advanced backcross QTL analysis for the identification of quantitative trait loci alleles from wild relatives of wheat ( Triticum aestivum L.). AB - Advanced backcross QTL (AB-QTL) analysis was used to identify quantitative trait loci (QTLs) for yield and yield components in a BC(2)F(2) population derived from a cross between the German winter wheat variety 'Prinz' and the synthetic wheat line W-7984 developed by CIMMYT. Two hundred and ten microsatellite markers were employed to genotype 72 pre-selected BC(2)F(2) plants and phenotypic data were collected for five agronomic traits from corresponding BC(2)F(3) families that were grown at four locations in Germany. Using single-marker regression and interval mapping, a total of 40 putative QTLs derived from W-7984 were detected, of which 11 were for yield, 16 for yield components, eight for ear emergence time and five for plant height. For 24 (60.0%) of them, alleles from the synthetic wheat W-7984 were associated with a positive effect on agronomic traits, despite the fact that synthetic wheat was overall inferior with respect to agronomic appearance and performance. The present study indicated that favorable QTL alleles could be transferred from wild relatives of wheat into an elite wheat variety for improvement of quantitative trait loci like yield by the advanced backcross QTL strategy and molecular breeding. To our knowledge, the results presented here were the first report on AB-QTL analysis in wheat. PMID- 12750780 TI - Analysis of HMW glutenin subunits and their coding sequences in two diploid Aegilops species. AB - Considerable progress has been made in understanding the structure, function and genetic regulation of high-molecular-weight (HMW) glutenin subunits in hexaploid wheat. In contrast, less is known about these types of proteins in wheat related species. In this paper, we report the analysis of HMW glutenin subunits and their coding sequences in two diploid Aegilops species, Aegilops umbellulata (UU) and Aegilops caudata (CC). SDS-PAGE analysis demonstrated that, for each of the four Ae. umbellulata accessions, there were two HMW glutenin subunits (designated here as 1Ux and 1Uy) with electrophoretic mobilities comparable to those of the x- and y-type subunits encoded by the Glu-D1 locus, respectively. In our previous study involving multiple accessions of Ae. caudata, two HMW glutenin subunits (designated as 1Cx and 1Cy) with electrophoretic mobilities similar to those of the subunits controlled by the Glu-D1 locus were also detected. These results indicate that the U genome of Ae. umbellulata and the C genome of Ae. caudata encode HMW glutenin subunits that may be structurally similar to those specified by the D genome. The complete open reading frames (ORFs) coding for x- and y-type HMW glutenin subunits in the two diploid species were cloned and sequenced. Analysis of deduced amino acid sequences revealed that the primary structures of the x- and y-type HMW glutenin subunits of the two Aegilops species were similar to those of previously published HMW glutenin subunits. Bacterial expression of modified ORFs, in which the coding sequence for the signal peptide was removed, gave rise to proteins with electrophoretic mobilities identical to those of HMW glutenin subunits extracted from seeds, indicating that upon seed maturation the signal peptide is removed from the HMW glutenin subunit in the two species. Phylogenetic analysis showed that 1Ux and 1Cx subunits were most closely related to the 1Dx type subunit encoded by the Glu-D1 locus. The 1Uy subunit possessed a higher level of homology to the 1Dy-type subunit compared with the 1Cy subunit. In conclusion, our study suggests that the Glu-U1 locus of Ae. umbellulata and the Glu-C1 locus of Ae. caudata specify the expression of HMW glutenin subunits in a manner similar to the Glu-D1 locus. Consequently, HMW glutenin subunits from the two diploid species may have potential value in improving the processing properties of hexaploid wheat varieties. PMID- 12750782 TI - Alloplasmic male-sterile Brassica juncea with Enarthrocarpus lyratus cytoplasm and the introgression of gene(s) for fertility restoration from cytoplasm donor species. AB - A new cytoplasmic male sterility (CMS) source in Brassica juncea (2n = 36; AABB) was developed by substituting its nucleus into the cytoplasm of Enarthrocarpus lyratus (2n = 20; E(l)E(l)). Male sterility was complete, stable and manifested in either petaloid- or rudimentary-anthers which were devoid of fertile pollen grains. Male sterile plants resembled the euplasmic B. juncea except for slight leaf yellowing and delayed maturity. Leaf yellowing was due mainly to higher level of carotenoids rather than a reduction in chlorophyll pigments. Female fertility in male-sterile plants varied; it was normal in lines having rudimentary anthers but poor in those with petaloid anthers. Each of the 62 evaluated germplasm lines of B. juncea was a functional maintainer of male sterility. The gene(s) for male-fertility restoration ( Rf) were introgressed from the cytoplasm donor species through homoeologous pairing between A and E(l) chromosomes in monosomic addition plants (2n = 18II+1E(l)). The percent pollen fertility of restored F(1) ( lyr CMS x putative restorer) plants ranged from 60 to 80%. This, however, was sufficient to ensure complete seed set upon by bag selfing. The CMS ( lyr) B. juncea compared favourably with the existing CMS systems for various productivity related characteristics. However, the reduced transmission frequency of the Rf gene(s) through pollen grains, which was evident from the sporadic occurrence of male-sterile plants in restored F(1) hybrids, remains a limitation. PMID- 12750783 TI - Genetic maps for Pinus elliottii var. elliottii and P. caribaea var. hondurensis using AFLP and microsatellite markers. AB - Genetic maps for individual Pinus elliottii var. elliottii and P. caribaea var. hondurensis trees were generated using a pseudo-testcross mapping strategy. A total of 329 amplified fragment length polymorphic (AFLP) and 12 microsatellite markers were found to segregate in a sample of 93 interspecfic F(1) progeny. The male P. caribaea var. hondurensis parent was more heterozygous than the female P. elliottii var. elliottii parent with 19% more markers segregating on the male side. Framework maps were constructed using a LOD 5 threshold for grouping and interval support threshold of LOD 2. The framework map length for the P. elliottii var. elliottii megagametophyte parent (1,170 cM Kosambi; 23 linkage groups) was notably smaller than the P. caribaea var. hondurensis pollen parent (1,658 cM Kosambi; 27 linkage groups). The difference in map lengths was assumed to be due to sex-related recombination variation, which has been previously reported for pines, as the difference in map lengths not be accounted for by the larger number of markers mapping to the P. caribaea var. hondurensis parent - 109 compared with 78 in P. elliottii var. elliottii parent. Based on estimated genome sizes for these species, the framework maps for P. elliottii var. elliottii and P. caribaea var. hondurensis covered 82% and 88% of their respective genomes. The pseudo-testcross strategy was extended to include AFLP and microsatellite markers in an intercross configuration. These comprehensive maps provided further genome coverage, 1,548 and 1,828 cM Kosambi for P. elliottii var. elliottii and P. caribaea var. hondurensis, respectively, and enabled homologous linkage groups to be identified in the two parental maps. Homologous linkage groups were identified for 11 out of 24 P. elliottii var. elliottii and 10 out of 25 P. caribaea var. hondurensis groups. A higher than expected level of segregation distortion was found for both AFLP and microsatellite markers. An explanation for this segregation distortion was not clear, but it may be at least in part due to genetic mechanisms for species isolation in this wide cross. PMID- 12750784 TI - Powdery mildew resistance gene Pm22 in cultivar Virest is a member of the complex Pm1 locus in common wheat ( Triticum aestivum L. em Thell.). AB - The powdery mildew resistance gene Pm22, identified in the Italian wheat cultivar Virest and originally assigned to wheat chromosome 1D, was mapped to chromosome 7A with the aid of molecular markers. Mapping of common AFLP and SSR markers in two wheat crosses segregating for Pm22 and Pm1c, respectively, indicated that Pm22 is a member of the complex Pm1 locus. Pm22 also showed a pattern of resistance reaction to a differential set of Blumeria graminis f. sp. tritici isolates that was distinguishable from those from other Pm1 alleles in lines Axminster/8*Cc ( Pm1a), MocZlatka ( Pm1b), Weihenstephan Stamm M1N ( Pm1c) and Triticum spelta var. duhamelianum TRI 2258 ( Pm1d). Based on these results, the gene symbol Pm1e is proposed for the powdery mildew resistance gene in cv. Virest. PMID- 12750785 TI - Dissection of resistance to soil-borne yellow-mosaic-inducing viruses of barley (BaMMV, BaYMV, BaYMV-2) in a complex breeders' cross by means of SSRs and simultaneous mapping of BaYMV/BaYMV-2 resistance of var. 'Chikurin Ibaraki 1'. AB - Ninety-three F(1)-derived doubled haploid (DH) lines from a complex breeders' cross involving the Japanese genotype 'Chikurin Ibaraki 1', which is resistant to barley mild mosaic virus (BaMMV) and two strains of barley yellow mosaic virus (BaYMV and BaYMV-2), three susceptible varieties ('Hamu', 'Julia' and a breeding line) and cv. 'Carola', which carries rym4 conferring resistance to BaMMV and BaYMV, were analysed for resistance to BaMMV, BaYMV and BaYMV-2. The DH lines fell into four phenotypic classes. In addition to completely resistant and susceptible genotypes, DHs were observed which were either resistant to BaMMV and BaYMV or to BaYMV and BaYMV-2. For BaMMV and BaYMV-2 resistance, segregation ratios approaching 1r:1s were observed, suggesting the presence of single resistance genes. In contrast, the segregation ratio for BaYMV fits a 3r:1s segregation ratio, suggesting the presence of two independently inherited genes. From the genetic analysis, we conclude that a resistance locus effective against BaYMV and BaYMV-2 originates from Chikurin Ibaraki 1 and segregates independently from the Carola-derived rym4 resistance that is effective against BaYMV and BaMMV. The BaMMV resistance in Chikurin Ibaraki 1 has probably been lost during population development. This hypothesis was tested using a simple-sequence repeat (SSR) marker (Bmac29) linked to rym4. All BaMMV-resistant DH lines supported amplification of the rym4-resistance diagnostic allele. To identify the genetic location of the Chikurin Ibaraki 1-derived resistance against BaYMV/BaYMV-2, bulked DNA samples were constructed from the four resistance classes, and bulked segregant analysis was performed using a genome-wide collection of SSRs. Differentiating alleles were observed at two linked SSRs on chromosome 5H. The location of this BaYMV/BaYMV-2 resistance locus was confirmed and further resolved by linkage analysis on the whole population using a total of five linked SSRs. PMID- 12750786 TI - Identification of S haplotypes in Brassica by dot-blot analysis of SP11 alleles. AB - A self-incompatibility system is used for F(1) hybrid breeding in Brassicaceae vegetables. The determinants of recognition specificity of self-incompatibility in Brassica are SRK in the stigma and SP11/SCR in the pollen. Nucleotide sequences of SP11 alleles are more highly variable than those of SRK. We analyzed the S haplotype specificity of SP11 DNA by Southern-blot analysis and dot-blot analysis using 16 S haplotypes in Brassica oleracea, and found that DNA fragments of a mature protein region of SP11 cDNA, SP11(m), of eight S haplotypes can detect only the SP11 alleles of the same S haplotypes. This specificity makes these methods useful for S haplotype identification. Therefore, we developed two methods of dot-blot analysis for SP11. One is dot blotting of DNA samples, i.e. plant genomic DNA probed with labeled SP11(m), and the other is dot blotting of SP11(m) DNA fragments probed with labeled DNA samples, i.e. the SP11 coding region labeled by PCR using a template of plant genomic DNA. The former is useful for testing many plant materials. The latter is suitable, if there is no previous information on the S haplotypes of plant materials. PMID- 12750787 TI - Identification of non-TIR-NBS-LRR markers linked to the Pl5/ Pl8 locus for resistance to downy mildew in sunflower. AB - The resistance of sunflower, Helianthus annuus L., to downy mildew, caused by Plasmopara halstedii, is conferred by major genes denoted by Pl. Using degenerate and specific primers, 16 different resistance gene analogs (RGAs) have been cloned and sequenced. Sequence comparison and Southern-blot analysis distinguished six classes of RGA. Two of these classes correspond to TIR-NBS-LRR sequences while the remaining four classes correspond to the non-TIR-NBS-LRR type of resistance genes. The genetic mapping of these RGAs on two segregating F2 populations showed that the non-TIR-NBS-LRR RGAs are clustered and linked to the Pl5/ Pl8 locus for resistance to downy mildew in sunflower. These and other results indicate that different Pl loci conferring resistance to the same pathogen races may contain different sequences. PMID- 12750788 TI - An intraspecific linkage map of the chickpea ( Cicer arietinum L.) genome based on sequence tagged microsatellite site and resistance gene analog markers. AB - An intraspecific linkage map of the chickpea genome based on STMS as anchor markers, was established using an F(2) population of chickpea cultivars with contrasting disease reactions to Ascochyta rabiei (Pass.) Lab. At a LOD-score of 2.0 and a maximum recombination distance of 20 cM, 51 out of 54 chickpea-STMS markers (94.4%), three ISSR markers (100%) and 12 RGA markers (57.1%) were mapped into eight linkage groups. The chickpea-derived STMS markers were distributed throughout the genome, while the RGA markers clustered with the ISSR markers on linkage groups LG I, II and III. The intraspecific linkage map spanned 534.5 cM with an average interval of 8.1 cM between markers. Sixteen markers (19.5%) were unlinked, while l1 chickpea-STMS markers (20.4%) deviated significantly ( P < 0.05) from the expected Mendelian segregation ratio and segregated in favor of the maternal alleles. However, ten of the distorted chickpea-STMS markers were mapped and clustered mostly on LG VII, suggesting the association of these loci in the preferential transmission of the maternal germ line. Preliminary comparative mapping revealed that chickpea may have evolved from Cicer reticulatum, possibly via inversion of DNA sequences and minor chromosomal translocation. At least three linkage groups that spanned a total of approximately 79.2 cM were conserved in the speciation process. PMID- 12750789 TI - Mapping of yield-related QTLs in pepper in an interspecific cross of Capsicum annuum and C. frutescens. AB - An advanced backcross QTL study was performed in pepper using a cross between the cultivated species Capsicum annuum cv. Maor and the wild C. frutescens BG 2816 accession. A genetic map from this cross was constructed, based on 248 BC(2) plants and 92 restriction fragment length polymorphism (RFLP) markers distributed throughout the genome. Ten yield-related traits were analyzed in the BC(2) and BC(2)S(1) generations, and a total of 58 quantitative trait loci (QTLs) were detected; the number of QTLs per trait ranged from two to ten. Most of the QTLs were found in 11 clusters, in which similar QTL positions were identified for multiple traits. Unlike the high percentage of favorable QTL alleles discovered in wild species of tomato and rice, only a few such QTL alleles were detected in BG 2816. For six QTLs (10%), alleles with effects opposite to those expected from the phenotype were detected in the wild species. The use of common RFLP markers in the pepper and tomato maps enabled possible orthologous QTLs in the two species to be determined. The degree of putative QTL orthology for the two main fruit morphology traits-weight and shape-varied considerably. While all eight QTLs identified for fruit weight in this study could be orthologous to tomato fruit weight QTLs, only one out of six fruit shape QTLs found in this study could be orthologous to tomato fruit shape QTLs. PMID- 12750790 TI - Genetic and physical mapping of a new gene for bacterial blight resistance in rice. AB - The inheritance of resistance for bacterial blight, caused by Xanthomonas oryzae pv. oryzae ( Xoo), was studied in Minghui 63, an elite restorer line for a number of widely used rice hybrids in China. A new dominant gene against a Chinese Xoo strain JL691 in both the seedling and adult stages was identified in Minghui 63 and designated as Xa26( t). Using a total of 477 highly susceptible individuals from an F(2) population, the Xa26( t) locus was mapped to a region of about 1.68 cM. This locus co-segregated with marker R1506 and was 0.21 cM from marker RM224 on one side and 1.47 cM from marker Y6855RA on the other side, in rice chromosome 11. A contig map, composed of five non-redundant bacterial artificial chromosome (BAC) clones and spanning approximately 500 kb in length, was constructed. Analysis of recombination events in the Xa26( t) region with the highly susceptible F(2) individuals anchored the gene locus to a region covered by three overlapped BAC clones. Assay of the lines showing a double crossover in marker loci flanking Xa26( t), in a population of recombinant inbred lines carrying Xa26( t), further delineated the gene to a 20-kb fragment. The Xa26( t) locus is tightly linked to another bacterial blight resistance gene locus, Xa4. PMID- 12750792 TI - Identification and mapping of a major gene (Ft1) involved in fructification time in the interspecific cross Coffea pseudozanguebariae x C. liberica var. Dewevrei: impact on caffeine content and seed weight. AB - Fructification time was studied in the interspecific cross Coffea pseudozanguebariae x C. liberica var. Dewevrei (PSE x DEW). Parental species, F(1) hybrids and offspring of the first backcross generation (BC(1)), consisting of F(1) x PSE (BCPSE) and F(1) x DEW (BCDEW) plants, were observed. Fructification time can be split into two independent visual phases: the full growth period, from blooming up to the end of fruit growth, and the maturation phase, defined by the green to red color change. Fructification time was found to be an additive trait. The full-growth period showed a bimodal distribution in the BCDEW hybrid, suggesting the involvement of Ft1, a major gene that was mapped on linkage group E. The main effects of Ft1 were to lower caffeine content and 100 seed weight, without any impact on chlorogenic acid, trigonelline and sucrose contents. Two molecular markers were identified that bracket Ft1 and which could be used for early marker-assisted selection. PMID- 12750791 TI - Comparative mapping of Phytophthora resistance loci in pepper germplasm: evidence for conserved resistance loci across Solanaceae and for a large genetic diversity. AB - Phytophthora capsici Leonian, known as the causal agent of the stem, collar and root rot, is one of the most serious problems limiting the pepper crop in many areas in the world. Genetic resistance to the parasite displays complex inheritance. Quantitative trait locus (QTL) analysis was performed in three intraspecific pepper populations, each involving an unrelated resistant accession. Resistance was evaluated by artificial inoculations of roots and stems, allowing the measurement of four components involved in different steps of the plant-pathogen interaction. The three genetic maps were aligned using common markers, which enabled the detection of QTLs involved in each resistance component and the comparison of resistance factors existing among the three resistant accessions. The major resistance factor was found to be common to the three populations. Another resistance factor was found conserved between two populations, the others being specific to a single cross. This comparison across intraspecific germplasm revealed a large variability for quantitative resistance loci to P. capsici. It also provided insights both into the allelic relationships between QTLs across pepper germplasm and for the comparative mapping of resistance factors across the Solanaceae. PMID- 12750793 TI - Mapping QTLs for seed dormancy and the Vp1 homologue on chromosome 3A in wheat. AB - A major component of the observed genetic variation for pre-harvest sprouting in wheat ( Triticum aestivum L.) appears to be the level of seed dormancy. Group 3 chromosomes have received attention as carrying the R genes for seed-coat color and the taVp1 genes that are orthologous to the maize Vp1 gene which encode a dormancy-related transcription factor. The objectives of the present study were to map quantitative trait loci (QTLs) for seed dormancy on chromosome 3A and to investigate an association between taVp1 or R-A1 and the QTLs detected. A mapping population in the form of recombinant inbred lines developed from the cross between the highly dormant Zenkoujikomugi (Zen) and Chinese Spring (CS) was utilized. Nineteen marker loci, including taVp1, were mapped on chromosome 3A. The taVp1 locus was located in the middle of the long arm, about 85 cM from the centromere. The population was evaluated in duplicate by growing them under controlled environment conditions. Two QTLs for seed dormancy, designated as QPhs.ocs-3A.1 and QPhs.ocs-3A.2, were identified on the short and long arms, respectively. QPhs.ocs-1 explained 23-38% of the phenotypic variation and the Zen allele had a striking effect on maintaining dormancy. QPhs.ocs-2, with a minor effect, was detectable only at the dormancy-breaking stage. Although QPhs.ocs-2 was loosely linked to taVp1 by around 50 cM, they are clearly distinct genes. Zen and CS carry the white R-A1a allele, and no QTL effect was detected in the vicinity region of R-A1. Hence it was concluded that the high dormancy associated with chromosome 3A of Zen is ascribable to QPhs.ocs-1 on the short arm but is not due to the direct contribution of either the taVp1 or R-A1 locus. PMID- 12750794 TI - AFLP analysis of genetic variability in New Guinea impatiens. AB - New Guinea impatiens ( Impatiens hawkeri) is an economically important floral crop, however, little work has been conducted to further our understanding of the genetics of this crop. In this study, we used amplified fragment length polymorphism (AFLP) technology to investigate the level of polymorphism present among 41 commercial cultivars of New Guinea impatiens, study their genetic relatedness, and assess the genetic diversity in this material. An efficient DNA extraction protocol was developed, and a total of 48 EcoRI and MseI primer combinations were used for PCR amplification. Amplification products were then subjected to polyacrylamide gel electrophoresis. The AFLP analysis showed that all 41 cultivars generated between 73 and 130 scoreable polymorphic bands per primer combination. Gower's Genetic Dissimilarity estimates for the entire set of cultivars ranged between 0.940 and 0.488. A dendogram was generated from these dissimilarity data that revealed four groupings among these 41 cultivars. The implications of these results on genotypic variation, genetic relationships, and genetic diversity in New Guinea impatiens will be discussed. PMID- 12750795 TI - Mapping QTLs for resistance against Globodera pallida (Stone) Pa2/3 in a diploid potato progeny originating from Solanum spegazzinii. AB - A "F1" diploid population between Solanum tuberosum 2 x and the wild Solanum spegazzinii was studied. It segregated for resistance against the potato cyst nematode Globodera pallida derived from the wild species. The inheritance had a quantitative nature. Linkage maps of AFLP and RFLP markers were constructed for both parents. Three QTLs were identified on the map of the resistant parent on chromosomes V, VI and XII, respectively. The first one had a major effect and explained more than 50% of the total variance of resistance. It is located in a cluster of resistance genes and may be the same locus as Gpa which has been described formerly. The two others explained about 20% of the total variance each. The QTL on chromosome XII is also in a cluster of resistance genes, and in an orthologous position with resistance genes against nematodes in tomato and pepper. PMID- 12750796 TI - Genetic and physical mapping of homologues of the virus resistance gene Rx1 and the cyst nematode resistance gene Gpa2 in potato. AB - Nine resistance gene homologues (RGHs) were identified in two diploid potato clones (SH and RH), with a specific primer pair based on conserved motifs in the LRR domain of the potato cyst nematode resistance gene Gpa2 and the potato virus X resistance gene Rx1. A modified AFLP method was used to facilitate the genetic mapping of the RGHs in the four haplotypes under investigation. All nine RGHs appeared to be located in the Gpa2/ Rx1 cluster on chromosome XII. Construction of a physical map using bacterial artificial chromosome (BAC) clones for both the Solanum tuberosum ssp. tuberosum and the S. tuberosum ssp. andigena haplotype of SH showed that the RGHs are located within a stretch of less than 200 kb. Sequence analysis of the RGHs revealed that they are highly similar (93 to 95%) to Gpa2 and Rx1. The sequence identities among all RGHs range from 85 to 100%. Two pairs of RGHs are identical, or nearly so (100 and 99.9%), with each member located in a different genotype. Southern-blot analysis on genomic DNA revealed no evidence for additional homologues outside the Gpa2/ Rx1 cluster on chromosome XII. PMID- 12750797 TI - [Molecular diagnostics of renal diseases with underlying genetic predisposition]. AB - The genetic basis for dysplasia and tumors of the kidney has increasingly become the subject of cytogenetic and molecular genetic investigations over the last decade. For that reason, it is now possible to define the risk of disease recurrence more precisely in families with kidney diseases caused by genetic alterations. The relevant genes and the mutations have been identified for most of these diseases and genetic diagnostics are possible. However, it is necessary to evaluate in each individual case whether genetic diagnostics are reasonable. This will be discussed for polycystic renal diseases, agenesis and dysplasia of the kidney, and hereditary kidney tumors. PMID- 12750798 TI - [Modern diagnostics for urological infections]. AB - This paper provides a short overview of modern, molecular-based diagnostic procedures of urogenital tract infections. Although gaining importance, molecular methods have not yet become a reliable substitution for the classic procedures in terms of costs and quality standards. As an example of a new molecular approach in microbiology, a method for the detection of the most relevant uropathogens in a single PCR is presented. Furthermore, the development of a real time PCR is described. PMID- 12750799 TI - [Molecular diagnostics of prostate cancer]. AB - Although a number of studies have identified molecular markers for prostate cancer, their clinical utility remains mainly unclear. Markers, which allow improved determination of the biological aggressiveness of individual prostate cancers, may help to optimize therapeutic management of this heterogeneous tumor type. Here, a subset of molecular markers, which are intensively discussed in the literature or which are supposed to gain clinical utility in the future, are described in more detail. For a better survey, the markers are divided into (a) susceptibility markers, (b) malignancy markers, and (c) aggressiveness markers. The number of markers described as well as the inconsistency across studies in assessing their clinical utility reflect the heterogeneity of prostate cancer also on a genetic level so that it is unlikely that a single marker will gain clinical relevance. Future research must include systematic analysis of the clinical utility of not only single markers but rather of marker profiles in appropriate studies. PMID- 12750801 TI - [Molecular diagnostics in urologic oncology. Detection of nucleic acids in urine samples]. AB - The goal of molecular diagnostics in oncology is the early diagnosis of malignant disease processes during initial work-up or as part of follow-up. Body fluids serve as the primary material for non-invasive diagnostic methods. Besides actual tumor cells, the examination of urine can yield evidence of secreted proteins or even free nucleic acids. In principle, all of the methods available for the detection of tumor markers in tissue or blood samples can be successfully applied to the examination of urine samples. However, molecular biological examination of urine samples is associated with important problems because the cells in such samples are exposed to significant degradation and regression effects and because certain components of the urine act to inhibit the polymerase chain reaction. The present overview discusses the respective strengths and weakness of the available technology as applied to the diagnosis of urologic malignancies. Experimental studies conducted to date have reported high sensitivities and specificities for molecular diagnostics using urine samples. It is important to note that not only carcinomas of the urinary bladder can be diagnosed from material obtained in urine samples: in fact, the method can be used to diagnose entities such as renal cell and prostate carcinomas and, due to renal filtration of DNA, even non urologic malignancies. The diagnostic application of these methods, however, remains in an experimental stage and must still clear several hurdles before becoming available for routine clinical use. PMID- 12750802 TI - [Chronic pain syndrome in uorology]. AB - Chronic pain of the genital region remains a therapeutic challenge. Among men, symptoms are mainly related to the prostate, bladder and scrotal organs, and among women to the bladder and vagina. Only some of the cases demonstrate pathologic changes of the symptomatic organs requiring specific treatment. Among pain medications, peripheral analgesics are the most suitable. In the case of chronic pain, which can be classified according to Gerbershagen, a psychosomatic origin also has to be considered and needs to be evaluated. Analgesics are of minor importance in the treatment of psychosomatic syndromes but tricyclic antidepressants or anticonvulsants may be helpful. Relaxation techniques also need to be considered. PMID- 12750800 TI - [Perspectives for molecular diagnostics exemplified by urothelial bladder carcinoma]. AB - The rapidly growing knowledge of molecular mechanisms will change the daily routine of clinicians in the near future. Regarding urothelial bladder carcinoma, one may expect that molecular diagnostics will identify patients susceptible to disease development by screening their genotype. Furthermore, in addition to histopathologic findings, prognostic markers will be used for disease management. In an ongoing multicenter trial, the decision on whether or not to treat patients with adjuvant chemotherapy after cystectomy is based on their p53 status. In the near future, cytostatic medications are expected to be chosen according to genetic profiles of the tumor or patient. New medications, which target tumor specific alterations of cell-signaling cascades in bladder or other cancers, prominently inhibitors of the ERBB membrane receptor family, are currently under clinical investigation and will undoubtedly form an important part of therapeutic oncologic regimens. In conclusion, evaluation of gene profiles of tumors and patients will gain importance for clinicians. PMID- 12750803 TI - [Defecation problems following radical perineal prostatectomy. A prospective study]. AB - PURPOSE: The true incidence of bowel symptoms prior to radical prostatectomy, as evaluated by a differentiated questionnaire, is unknown. We therefore performed a prospective study on patients who were scheduled for radical perineal prostatectomy. MATERIAL AND METHODS: A total of 67 patients with cT1-cT3, N0, M0 prostate cancer underwent an extrafascial, radical, perineal prostatectomy. The patients received a questionnaire prior to surgery as well as 6 months and 12 months after surgery. This took into account demographic data, stool symptoms (Kelly questionnaire) and questions concerning bladder function (ICS male continence questionnaire). The questionnaires were evaluated by three of the authors not involved in patient care. RESULTS: The mean age of the patients was 64 years. The PSA range was 0.9-55.6 ng/ml (mean 12.7 ng/ml). There were 47 pT2, 19 pT3 and a single pT4 tumour. A total of 59 patients had a Gleason score of 6 or less. Positive surgical margins were present in four patients. The 12 months follow-up questionnaire could be evaluated for 82% of the patients (n=55). In addition, 46 patients answered the questionnaire at 6 months post-surgery. Three or more problems in relation to bowel movements were reported preoperatively by 21% of the patients. Straining with bowel emptying was the symptom which was indicated most often. Stool smearing was reported by 13% of patients at least once or twice monthly. In addition, 6% of patients reported that they had difficulties in differentiating soft stool from gas. After 1 year, seven (13%) of the patients reported stool smearing which was not present prior to surgery. Six of these patients observed this problems only once or twice a month. Only one patient had stool smearing once a week. Two patients reported urgency, two had a decreased warning time and one reported decreased sensibility. Two patients used protective pads. The most frequent symptom reported postoperatively was straining with bowel emptying (24% preoperatively and 16% postoperatively). CONCLUSION: It is evident that men scheduled for radical prostatectomy already have significant stool problems preoperatively. Newly developed, postoperative stool smearing on a daily basis occurred extremely seldom. They same is true for the discrimination between soft stool and gas. From our point of view, it is important to use the extrasphincteric approach to the prostate described by Young. Furthermore, the pubo-anal sling should be preserved. PMID- 12750804 TI - [Can nomograms derived in the U.S. applied to German patients? A study about the validation of preoperative nomograms predicting the risk of recurrence after radical prostatectomy]. AB - In patients suffering from prostate cancer, preoperative nomograms, which predict the risk of recurrence may provide a helpful tool in regard to the counselling and planning of an appropriate therapy. The best known nomograms were published by the Baylor College of Medicine, Houston and the Harvard Medical School, Boston. We investigated these nomograms derived in the U.S. when applied to German patients. Data from 1003 patients who underwent radical prostatectomy at the University-Hospital Hamburg were used for validation. Nomogram predictions of the probability for 2-years (Harvard nomogram) and 5-years (Kattan nomogram) freedom from PSA recurrence were compared with actual follow-up recurrence data using areas under the receiver-operating-characteristic curves (AUC). The recurrence free survival after 2 and 5 years was 78% and 58%, respectively. The AUC of the Harvard nomogram predicting 2-years probability of freedom from PSA recurrence was 0.80 vs. Kattan-Nomogram 5-years prediction of 0.83. Thereby, the Kattan nomogram showed a significant higher predictive accuracy (p=0.0274). For that reason preoperative nomograms derived in the U.S. can be applied to german patients. However, we would recommend the utilization of the Kattan nomogram due to its higher predictive accuracy. PMID- 12750806 TI - [Analgesic compress with Instillagel following circumcision. Observations]. PMID- 12750805 TI - [Angiomyolipoma of the kidneys as a rare cause of retroperitoneal hemorrhage. Two case reports with tuberous sclerosis Bourneville-Pringle]. AB - Tuberous sclerosis (Bourneville-Pringle-disease, TSC) is an autosomal dominant disorder characterized by seizures, mental retardation and hamartomatous tumours in multiple organs, including subependymal giant cell astrocytomas, cardiac rhabdomyomas and renal angiomyolipomas. Recent population-based studies suggest a prevalence of 1 case per 25,000 individuals. Renal angiomyolipomas, which may be found sporadically or associated with TSC, become evident as an acute retroperitoneal haemorrhage or by symptoms of a flank mass. Ultrasound and computed tomography provide clear evidence of lipomatous formation while, in rare instances, angiography can demonstrate the existence of multiple vascular tumour compartments. In view of two cases which were admitted with the clinical picture of an acute abdomen on the basis of retroperitoneal haemorrhage, the therapeutic strategies for TSC patients with renal angiomyolipomas are discussed, paying regard to the literature in this field. PMID- 12750808 TI - [The role of the tibiofibular syndesmotic and the deltoid ligaments in stabilizing Weber B type ankle joint fractures--an experimental investigation]. AB - The purpose of the present biomechanical investigation was to check the functional importance of the syndesmosis ligaments and of the deltoid ligament for ankle fracture type B according to the AO-Weber classification. We constructed a special fixation clamp, with 12 fresh and unembalmed lower legs being tested for lateral shift (mm) and ten for tibiotalar rotation. All specimens were exposed in the same neutral position. Transverse loads (F(y)) varied between 0 and 150 N, axial loads (F(z)) between 0, 300, 600 and 1,000 N and rotational loads (F(r)) between 2.4 and 4.9 Nm. All series were repeated according to supination-eversion (SE) injury patterns of the Lauge-Hansen classification. Syndesmotic ligaments and the fibula were incrementally sectioned from anterior to posterior. Type SE I consisted of an isolated incision of the anterior syndesmosis ligament. Type SE II had an additional oblique fracture of the fibula at the height of the tibiofibular syndesmosis. In type SE III injuries, in addition to the fibular fracture, a complete rupture of the syndesmosis ligaments was present, and for type SE IV lesions the deltoid ligaments were incised. The transverse load-displacement curve was s-shaped in all uninjured joints,with the highest gradient between 10 and 20 N with no axial compression. Without axial compression in cases of F(y)=25 N transverse loads, the mean talus translation was 0.51 mm. Following type II injuries, the average talus translation was 0.68 mm (not significant) and rose to an average of 0.95 mm ( P <0.01) in type III injuries. After additional incision of the deltoid ligaments, the ankle joint subluxed permanently when more than 5-10 N transverse loads were applied. Axial loads of 300 N or more resulted in a considerable reduction in talus translations, indicating increased stability and congruency within the joint complex. In this way, the vertical loading of the ankle joints always contributed to joint stability. The average internal tibiotalar rotation reached with a torque of 2.4 Nm was 3.52 degrees and with 4.9 Nm 5.15 degrees when no axial compression was applied. External rotation measured -6.36 degrees and -8.62 degrees, respectively. Following the experimental protocol, significant increases were noted for external rotation at SE II degrees injuries ( P =0.003) and for internal rotation at SE III degrees ( P =0.03) injuries. Our data support the proposition that the deltoid ligaments and the posterior syndesmosis play a key role in the stability of ankle fractures for supination-eversion injuries. If these structures remain intact, conservative and early functional treatment are recommended in patients with minimal (<2 mm) or no fracture displacement. This concept is confirmed by the literature dealing with clinical mid- and long-term follow-up studies. PMID- 12750807 TI - [Guidelines of German urologists on therapy of benign prostate syndrome]. PMID- 12750809 TI - [A bipolar radial head prosthesis after comminuted radial head fractures: indications, treatment and outcome after 5 years]. AB - The treatment of comminuted fractures of the radial head with concomitant injuries of the ulnar complex by resection of the radial head usually does not provide satisfactory long-term results. Other than joint instability in the elbow and a limited range of motion, radius proximalisation in the sense of ulnocarpal impingement, osteoarthritis and pain in the elbow have been described. Between 1995 and 1997, 11 radial head prostheses were implanted in ten patients who had sustained a comminuted fracture of the radial head with concomitant injury to the ulnar complex. A follow-up survey was conducted with the patients on average 5 years after the injury. Eight patients with nine implants participated in the follow-up, one patient had died and another refused to participate but declared that he did not suffer from any impairment. According to the Morrey score, two of the results were found to be very good, five to be good, one to be fair and one to be poor. Despite the severe injuries sustained by the elbow, neither joint instability in the elbow nor proximalisation of the radius, cubitus valgus, ulnar nerve syndrome, nor loosening of the prosthesis were found in any of the patients. In the event of comminuted fractures of the radial head which are impossible to reconstruct by osteosynthesis and which occur with concomitant ulnar ligamentous or osseous injury, the implantation of a prosthesis is preferred over the resection of the head of the radius. PMID- 12750810 TI - [Operative or conservative treatment of anterior cruciate ligament rupture: a systematic review of the literature]. AB - The aim of this paper was to examine all studies that compared operative and conservative treatment of anterior cruciate ligament (ACL) rupture. Covering a period of 22 years all relevant articles were retrieved from the medical databases Medline, Embase, and Cochrane. The results of these studies were judged on to the criteria of subjective stability (giving-way phenomenon), objective stability (pivot shift test), and return to the preinjury level of sports. There were 11 different studies published in 16 articles consisting of 2 randomized,2 prospective, and 7 retrospective trials. Except in one study the results in the operative groups were always better than in the conservative groups. The meta analysis of both randomized studies showed significant advantages for the operative treatment. Although arthroscopic ligament plasty is an established standard method for the treatment of ACL rupture, there is almost no basis for this procedure in the literature.Thus,new clinically applicable studies are necessary to develop objective guidelines for the treatment of anterior cruciate ligament tears. PMID- 12750811 TI - [Emergency room patients]. AB - The aim of the study was to examine the predictive value of CT for severe intracranial lesions of mild (GCS 15-13) and moderate (12-9) head injuries. Further,we examined the possibility of predicting these lesions by various variables/factors. Data were collected prospectively from the trauma registry of the DGU (Deutsche Gesellschaft fur Unfallchirurgie). Patients with a GCS score from 15-13 and from 12-9 were included in this study and examined for intracranial lesions (AIS(head) 3-6). Over a time period from 1993 to 1999, 1778 patients with mild head injury and 235 patients with moderate head injury were analyzed. Severe intracranial lesions were suffered by 18.6% of the patients with mild head injury and 50.4% of the patients with moderate head injuries. Of the predictive variables, heart rate,patients' age,and primary assessment by the emergency physician showed a strong correlation with the later observed intracranial lesions. IN CONCLUSION: (1) independently of the initially good GCS, a high percentage of patients suffered from severe intracranial lesions and (2) besides the GCS only the patients'age and primary assessment by the emergency physician were useful for identifying patients at risk for an intracranial lesion. PMID- 12750812 TI - [Ultrastructural examination of cell-mediated degradation of a calcium phosphate ceramic]. AB - Up to now little is known about cell-mediated degradation of biomaterial surfaces, especially as methodologically significant studies are not available. Therefore, the present study focused on ultrastructural details of cells involved in degradation of calcium phosphate ceramics. For the experimental procedure six adult sheep were used. At the medial aspect of the left hindlimb a cylindrical defect was created at the level of the proximal epiphysis of the tibia. Subsequently, a calcium phosphate paste was packed into the defect. Six weeks after implantation, specimens from the implants fixed by perfusion were examined histologically and by transmission electron microscopy. The results of light microscopy revealed substitution of the ceramic by newly formed lamellar bone. Electron microscopy indicated multinucleated cells localized at the implantation site and the bone surface, corresponding to osteoclasts. They formed resorption lacunae and revealed typical ultrastructural features such as the ruffled border and the sealing zone. Osteoclast-mediated degradation was performed by simultaneous resorption and phagocytosis. For the first time this degradation mechanism was documented in vivo. It confirms the notion that osteoclasts are members of the monocyte/macrophage family. PMID- 12750813 TI - [Risk factors for the development of pneumonia in multiple injured patients. Results of a prospective clinical trial]. AB - Pneumonia is the most common infectious complication in multiple trauma patients. In a prospective clinical cohort study, 266 multiply injured patients were examined for the development of pneumonia. Various risk factors were tested in uni- and multivariate analyses. Three different definitions of pneumonia were used in order to examine how results depended on definition. The incidence of pneumonia was 41%, but varied with definition (30-50%). Injuries to the thorax, head,and abdomen were associated with a significantly increased risk of pneumonia (adjusted relative risk: 1.77, 1.97,and 1.52, respectively).Furthermore, increasing age led to a higher risk of pneumonia. Although the primary analysis revealed a higher pneumonia risk in male patients (adjusted relative risk: 2.23; 95% CI: 1.43-3.05), this result could not be consistently reproduced when using other definitions of pneumonia. Trunk and head injuries and age are proven risk factors for developing posttraumatic pneumonia. The association between male gender and an increased rate of infectious complications remained questionable. PMID- 12750815 TI - [Proximal femur fracture with secondary hyperparathyroidism]. AB - We rarely see trauma patients with secondary hyperparathyroidism. We report about a case of a 68-year-old African female with a pertrochanteric femur fracture after a falling accident. As a secondary result we noted a Looser zone (pseudofracture) in the fractured femur. Extended diagnostic work-up including determination of vitamin D metabolites indicated the existence of a secondary hyperparathyroidism. The dark tanned phenotype of the African patient in combination with usually less solar radiation in central Europe seems to be the causative factor for the secondary hyperparathyroidism. Thus, this resulted in reduced cutaneous vitamin D synthesis. The fracture of the proximal femur was stabilized by osteosynthesis with a gamma nail; we thereby bridged the Looser zone. PMID- 12750814 TI - [Ipsilateral femoral fracture and anterior hip dislocation in a 15-year-old girl: case report]. AB - Operative management of concomitant ipsilateral femur fracture and anterior hip dislocation has not been previously described in the literature. We report the case of a 15-years-old girl who was injured in a motorcycle accident and presented with a femoral shaft fracture and a concomitant ipsilateral anterior hip dislocation. Operative management consisted of an attempted closed reduction of the femoral fracture, which was unsuccessful; thus, an external fixator was temporarily applied. Subsequently, the hip dislocation was treated by open reduction through an anterolateral approach. Finally, the femoral fracture was securely stabilized using an unreamed femoral intramedullary nail. The postoperative course was uneventful. MRI follow-up after 6 weeks did not reveal any sign of mangled vascularization of the femoral head and radiographs demonstrated normal bony healing of the shaft fracture. Due to the fact that there is no definitive surgical strategy for this rare combination of injuries, concomitant ipsilateral femoral fracture and anterior hip dislocation is an interesting and challenging situation for the trauma surgeon which requires a subtle and exact surgical technique in order to achieve satisfying results. PMID- 12750816 TI - [Surgical treatment of an acetabular fracture during pregnancy]. AB - We report about a case of a pregnant women in the 23rd gestation week who sustained an isolated acetabular fracture in a car accident. The fracture was treated surgically by open reduction and internal fixation 6 days after trauma. The outcome for the mother and the baby was excellent; both could be followed up for 1.5 years. The baby did not suffer from any disease related to the diagnostic or surgical procedures. We conclude from this case and from reviewing the literature that the operative fixation of an acetabular fracture during pregnancy is the appropriate treatment with minimal risk for the unborn child and best outcome for the mother. PMID- 12750817 TI - [Sigmoid colon perforation with local peritonitis caused by indirect trauma--case report and review of the literature]. AB - Injuries to the sigmoid occur either as acute or protracted events. In the first case, enteral contents discharge into the abdominal cavity and a generalized, fecal, life-threatening peritonitis with a bad prognosis develops. In the protracted form, the rupture is covered by peritoneum and adherent organs before perforation. The ensuing abscess formation may lead to perforation into contiguous visceral organs or the cutis. Frequently an intestinal or cutaneous fistula results. The trigger for a sigmoid perforation can be a spontaneous rupture in an already vulnerable intestine. Common precursory diseases are diverticulitis, colitis, carcinomas, and necroses. Also, elevated intestinal pressure invoked by increased bearing down or coproliths may cause disruption. Diagnostic procedures such as rectoscopy and rectal contrast instillation are frequent idiopathic causes of traumatic injuries to the sigmoid. Perforating injuries of the abdominal cavity by stabbing, gunshot, or impalement may affect the sigmoid and open its lumen. Foreign bodies often lead to traumatic injuries of the rectosigmoid junction. In contrast, indirect trauma as a cause of sigmoid perforation, which is described in the following case, is very rare. A 62-year old woman,who had a cholecystectomy and adhesive strangulation of intestine in her history, was admitted to our clinic after falling down stairs and landing on her bottom. She suffered a sigmoid rupture and peritonitis. Laparotomy and suturing of the sigmoid defect were performed. PMID- 12750818 TI - [OR-manager: surgeon or anaesthetist?]. AB - The heart of any surgical department is the operating room (OR) area. Any disturbances in the daily routine will affect the work flow of the whole hospital. On account of its central function, with numerous connections to other departments, the OR is the crystallisation point for deficiencies in various events and processes. As an example, the major complaints made by a surgical department regarding workflow and communication are outlined. To solve these problems, an "OR organization" team was established, which worked on the basis of a newly developed OR statute. Within 1 year the employees were more contentment and the workflow had improved. However, even in the second year of central OR management there is still the need to further stabilize the system as mismanagement still occurs. PMID- 12750819 TI - Allele-specific differential expression of a common adiponectin gene polymorphism related to obesity. AB - Adiponectin gene polymorphisms have recently been reported to be associated with obesity, insulin sensitivity, and the risk of type 2 diabetes. We examined a T94G polymorphism of the adiponectin gene in 245 ostensibly normal nondiabetic subjects. The G allele frequency was lower among subjects with higher BMI (> or =27) than in those with lower BMI. BMI was inversely correlated with the dose of G allele. Multivariate linear regression analyses showed that the adiponectin genotypes were significantly related to BMI after adjusting for age and gender. The dose of the G allele was associated with a reduction of approximately 1.12 kg/m(2) in BMI. We further found that the relative mRNA levels of G allele were consistently higher than those of T allele in the omental adipose tissue from 21 heterozygous subjects. Finally, we observed that the expression levels of adiponectin affected insulin-stimulated glucose uptake in differentiated 3T3-L1 adipocytes. In conclusion, the allele-specific differential expression of this common polymorphism could be responsible for its biological effects observed in this and the other studies. PMID- 12750820 TI - The long QT interval is not only inherited but is also linked to cardiac hypertrophy. AB - This review focuses on the molecular determinants of the duration of the QT interval as measured on by electrocardiography in normal subjects and during cardiac hypertrophy and failure. (a) In control conditions, on a single cell, the shape and duration of the action potential is the result of a balance between different ion currents which in turn were determined by the number of functional channels. On multicellular preparations the QT duration also represents the repolarization time; nevertheless it is modified by the transmural gradients. On body-surface electrocardiography the duration of the QT interval depends also of an additional factor: the spatial three-dimensional projection of the electrical waves vectors, which makes any determination of the epicardial dispersion by measuring QT interval dispersion questionable. (b) The enhanced action potential duration is well documented in cardiac hypertrophy and heart failure and is usually caused by a reduction in outward current densities in most of the species except mice. Among these currents I(tO) is the most frequently altered, especially in humans. Such an altered current density is caused by a diminished expression of the genes encoding either the ion channel subunits or regulatory proteins, such as KChIP2. In addition, hypertrophy modifies or even reverses the transmural gradient. In human and rats hypertensive cardiopathy is associated with a prolongation of the QT interval duration. The reduction in I(tO) is likely to be adaptive; it participates in the slowing of the cardiac cycle and reflects the fetal genetic reprogramming. Recent data also suggest that a reduction in the transient outward K(+) current density triggers protein synthesis through an activation of the calcineurin pathways. Thus a prolongation of the QT interval is not only inherited or drug-induced; it is also an essential component of the adaptive process in chronic mechanical overload. It is fundamentally incorrect to measure QT dispersion on a surface electrocardiography, but the mean QT interval may provide information concerning the progression of the disease, just as, and with the same restrictions, in the case of the quantification of V(max). PMID- 12750822 TI - [Comparison of algorithms for management of the difficult airway]. AB - Management of the difficult airway and maintenance of the oxygenation are the most important tasks of the anaesthetist. Respiratory problems are still the most important single cause for anaesthesia-related accidents with poor outcome. Algorithms are step-wise procedures developed from a great number of recommendations and are well suited to automation and training procedures. There is strong agreement among consultants that specific strategies lead to improved outcome, although, strictly speaking the degree of benefit on airway management cannot be clearly determined. Several anaesthesia societies, including the American Society of Anesthesiology,have developed their own algorithms for management of the difficult airway. The comparison of published algorithms shows that the management of the anticipated difficult airway has to be performed in the awake patient and fiberoptic intubation is a crucial part of that procedure. There are different techniques (different blades, guide wire, laryngeal mask, fiber optics) for the management of the unanticipated difficult airway. The laryngeal mask, transtracheal access and the Combitube are recommended for the management of the cannot intubate, cannot ventilate situation. More important than the questions which algorithm, which technique and which instruments should be used,is that each department has and practices its own algorithm. This strongly depends on local circumstances and personal preferences. Daily practice is the condition for the successful use in an emergency situation. The management is easier if one uses a simple algorithm and as few instruments as possible. PMID- 12750821 TI - Rett syndrome: the complex nature of a monogenic disease. AB - Rett syndrome (RTT) is a severe neurodevelopmental disorder affecting almost exclusively girls. It is currently considered a monogenic X-linked dominant disorder due to mutations in MECP2 gene, encoding the methyl-CpG binding protein 2. A few RTT male cases, resulting from mosaicism for MECP2 mutations, have been reported. Male germline MECP2 mutations cause either severe encephalopathy with death at birth (usually in brothers of classical RTT females) or X-linked recessive mental retardation (XLMR). To date the wide phenotypic heterogeneity associated with MECP2 mutations in females (from classical RTT to healthy carriers) has been explained by differences in X chromosome inactivation. However, conflicting results have been obtained in different studies, with both random and highly skewed X-inactivation reported in healthy carrier females. Consequently it is possible that mechanisms other than X-inactivation play a role in the expressivity of MECP2 mutations. To explain the phenotypic heterogeneity associated with MECP2 mutations we propose a digenic model in which the presence of a "mutated" allele in a second gene, leading to a less functional protein, determines the clinical severity of the MECP2 mutation. The model is supported by the identification of the same mutation in XLMR and RTT cases. The carrier mothers of XLMR families are clinically asymptomatic and present balanced X chromosome inactivation. Therefore the same mutation arising in different genetic backgrounds can cause XLMR in males, remain silent in the carrier females and cause classic RTT in females. MECP2 mutations account for approximately 70-80% of classic RTT cases. MECP2 negative cases might result from mutations in noncoding regions of MECP2 gene. Alternatively, these cases might be due to mutations in other genes (locus heterogeneity). This hypothesis is supported by the identification of several chromosomal rearrangements in MECP2 negative patients with RTT and RTT-like phenotypes. MeCP2 is considered a general transcriptional repressor. However, conditional mouse mutants with selective loss of Mecp2 in the brain develop clinical manifestations similar to RTT, indicating that MECP2 is exclusively required for central nervous system function. The involvement of MeCP2 in methylation-specific transcriptional repression suggests that MECP2 related disorders result from dysregulated gene expression. Studies on gene expression have been performed in mouse and human brains. A relatively small number of gene expression changes were identified. It is possible that MeCP2 causes dysregulation of a very small subset of genes that are not detected with this method of analysis, or that very subtle changes in many genes cause the neuronal phenotype. PMID- 12750823 TI - [A population-based survey of critical care]. AB - OBJECTIVES: Results of a population-based survey of critical care in a German state are presented. The indication for critical care was derived using the APACHE-II score,which measures the physiologic state of patients, and the TISS-28 score as a measure for expenditure of critical care. METHODS: During 3 months all adult patients of intensive care units in Saarland, a German state with 1.08 million residents,were included. All kinds of hospitals were involved. Completeness and validity of data are proved. RESULTS: The TISS score was registered daily on 13,391 adult patients with 54,503 days, the APACHE score during admission and discharge of intensive care units only. The correlation between the scoring-systems and indication, kind of care (critical care vs. intermediate care), kind of hospital, and ICD-10 diagnoses is analysed in this paper. CONCLUSIONS: In this study the population-based critical care in a state is analysed the first time. It completes previously published studies,which were performed at the level of hospitals and departments. PMID- 12750824 TI - [Incompatibility reactions in the intensive care unit. Five years after the implementation of a simple "colour code system"]. AB - In intensive care units (ICU) most of the drugs have to be administered by y piece infusions or admixtures. Drug stability and compatibility are critical elements in the accurate and appropriate delivery of drug therapies to patients. Five years after the implementation of a simple "colour code system" in an attempt to minimize the number of incompatibilities, the situation has been re examined. The clinical pharmacist collected 78 different medication regimes and the compatibility and incompatibilities were evaluated based on the available literature. Before initiating the "colour code system" in the ICU, 15% of the administered drugs were incompatible and afterwards the number decreased to 2%. This rate could be kept at 2%, even 5 years after the implementation of the system,without any further intervention. As a result of teamwork between nurses, doctors and clinical pharmacy a simple "colour system" was established to minimize incidences of drug incompatibility in the ICU. The system is highly accepted because the degree of uncertainty has been considerably reduced. PMID- 12750825 TI - [The Lazarus phenomenon. Spontaneous return of circulation after unsuccessful intraoperative resuscitation in a patient with a pacemaker]. AB - We present a case of spontaneous recovery after failed intraoperative cardiopulmonary resuscitation in a patient with a cardiac pacemaker. Of the various mechanisms discussed in the literature, that which seems most relevant in our case of a Lazarus phenomenon, is that of impeded venous return in the course of positive pressure ventilation and hypovolemia.With the disconnection of ventilation, passive cardiac filling combined with electrical stimulation due to the cardiac pacemaker may have caused spontaneous myocardial electrical activity. Corresponding to previous authors,we recommend continued monitoring for 10 min after cessation of cardiopulmonary resuscitation. In addition, we propose a further attempt by disconnecting ventilation and external myocardial stimulation in the case of unresponsiveness to resuscitation efforts. PMID- 12750826 TI - [Pathophysiological and therapeutic aspects of amniotic fluid embolism (anaphylactoid syndrome of pregnancy): case report with lethal outcome and overview]. AB - A 35-years old gravida IV and para II underwent caesarean section because of fetal distress following induction of labour. During operation the patient developed disseminated intravascular coagulation (DIC), severe haemorrhage and shock necessitating massive blood transfusion,hysterectomy with pelvic packing, and high-dose catecholamines. Ultimately, recombinant factor VIIa was given to control bleeding. During the first 24 hours after operation, both clinical and laboratory findings showed that the severe DIC was on the course to recovery.However, the patient subsequently developed multiple organ dysfunction syndrome with respiratory and renal failure requiring mechanical ventilation and haemodialysis.All therapeutical efforts could not help that the patient passed away due to an inevitable multiple organ failure on the 12th day after the operation. Given the constellation of diagnostic and clinical findings, the most likely diagnosis was amniotic fluid embolism (AFE), a rare complication of pregnancy. The following differential diagnoses were less likely or excluded in this reported patient: pre-eclampsia/pregnancy-induced hypertension,HELLP syndrome,anaphylaxis,uterine rupture, transfusion reactions,pulmonary embolism. AFE occurs rarely, and because studies in animal models cannot reproduce accurately the pathophysiological and clinical alterations seen in humans, its pathogenesis remains unclear. It has been proposed that the clinical syndrome of AFE occurs when fetal antigens pass the maternal immunological barrier in susceptible mothers. The recognition of fetal antigens by maternal immune system subsequently triggers the release of endogenous mediators that are responsible for dramatic pathophysiological disturbances.Furthermore, the components of amniotic fluid initiate the DIC. These events are more consistent with septic shock and anaphylactic shock than with an embolic process and it was proposed that the term "amniotic fluid embolism" be changed to "anaphylactoid syndrome of pregnancy". At present, no therapy has been found to consistently improve outcomes in women with AFE.Patients who survive the initial insult are at high risk for multiple organ failure. The mortality of AFE remains high. PMID- 12750827 TI - [Application of muscle relaxants in Germany. A survey of German anaesthesia departments]. AB - AIM: The aim of this study was to evaluate the use and application of muscle relaxants and neuromuscular monitoring in Germany. METHODS: A total of 2,996 questionnaires were sent out to the heads of German anaesthesia departments and private anaesthesia practices. The questions covered frequency of muscle relaxants used,how they were used, and neuromuscular monitoring. Influences on the way muscle relaxants were used could be derived from the desire for specific properties of a muscle relaxant, the desire for different monitoring conditions and from the size of the institution. We correlated these features with application practice using logistic regression analyses. RESULTS: Of the 2,996 questionnaires 2,058 could be analysed (68.6%). Amongst those were 102 level one hospitals (5%) and 903 private practices (44%). The replies from 350 (17%) departments were based on surveyed data, 1,613 (78.5%) were based on estimations. The desire for certain properties of muscle relaxants correlated with the use in practice, as were the desire for a non-depolarizing replacement for succinylcholine, the size of the department and the frequency of use of neuromuscular monitoring. Over 50% of all German anaesthesia departments limited the use of muscle relaxants to three. The use of laryngeal masks reduced the use of muscle relaxants. CONCLUSIONS: The survey regarding use of muscle relaxants in Germany could for the first time give an overview on the use of anaesthesia specific substances in Germany. From the different frequencies of use and use modalities,conclusions could be drawn towards a standard of application for the year 2000. Changes in this standard would raise the need for further trend surveys. The methods of statistical analysis and survey evaluation can be used as a base for further surveys. PMID- 12750828 TI - [The use of muscle relaxants for routine induction of anesthesia in Germany]. AB - The aim of this study was to evaluate the use of muscle relaxants during induction of anesthesia in patients without risk of aspiration of stomach contents. Of the 2,996 questionnaires sent out, 2,054 (68.6%) could be analysed and the results show that succinylcholine is used regularly in 13.6% of anesthesia departments. The next most commonly used muscle relaxants are atracurium, vecuronium and mivacurium, followed by cis-atracrium, rocuronium and pancuronium. Alcuronium is the least frequently used muscle relaxant. During induction of an elective anesthesia procedure, a priming technique is used by 19% of anesthesiologists, 22% utilize precurarization, and a timing technique is performed in 7.1%. The use of muscle relaxants for on-going relaxation follows the same pattern as for induction of neuromuscular blockade and succinylcholine is used in 1.4% if further relaxation is needed. The desire for specific qualities of muscle relaxants is correlated with higher use of the specific substance: short onset time for rocuronium, good controllability with mivacurium, no side-effects with cisatracurium and economical aspects with alcuronium. Of the participants 76.6% voiced the desire for a non-depolarizing replacement for succinylcholine.Private practices use mivacurium more often than hospitals, level one hospitals use rocuronium and cisatracurium more often. This survey could not show a definite standard of use in terms of muscle relaxants for an elective case.Precurarization, priming and timing are used frequently in patients not at risk of aspiration. This should be reduced by on-going teaching. PMID- 12750829 TI - [Opioids and immunosuppression. Clinical relevance?]. AB - First observations that opioids may have disadvantageous effects on the immune response have been made more than 100 years ago. Today the immunosuppressive effect of morphine is well established.Drug-induced immunomodulation is of growing importance in modern anesthetic concepts. The reduced stress response observed after morphine application contributes to this effect as well as direct impairment of immune effector cells such as bactericidal activity, intracellular killing,proliferative response or cytokine synthesis. Opioid-induced immunomodulation is mediated by opioid receptors found on immunocytes and in the central nervous system.A negative feedback mechanism via the hypothalamo pituitary-adrenal axis may potentiate the direct inhibitory effect of morphine on the immune response.A final statement regarding the clinical relevance of opioid induced immunosuppression cannot be made at this point, since the existing clinical data are preliminary and inconclusive.Therefore, further clinical studies are mandatory to elucidate the influence of opioid treatment on immune regulation in different clinical settings in anesthesia, critical care, pain therapy and emergency medicine. Further investigations may help to not only provide sufficient analgesia by application of opioids, but also to assess advantages and disadvantages on immune function. PMID- 12750830 TI - [Hepatitis C. The point of view of a clinical anaesthetist and intensive care specialist]. PMID- 12750831 TI - [Hepatitis C virus infection 2003 from the point of view of a hepatologist]. PMID- 12750832 TI - [Hepatitis C from the stand point occupational medicine]. PMID- 12750833 TI - Progression age enhanced backward bifurcation in an epidemic model with super infection. AB - We consider a model for a disease with a progressing and a quiescent exposed class and variable susceptibility to super-infection. The model exhibits backward bifurcations under certain conditions, which allow for both stable and unstable endemic states when the basic reproduction number is smaller than one. PMID- 12750834 TI - A mathematical model of cell-to-cell spread of HIV-1 that includes a time delay. AB - We consider a two-dimensional model of cell-to-cell spread of HIV-1 in tissue cultures, assuming that infection is spread directly from infected cells to healthy cells and neglecting the effects of free virus. The intracellular incubation period is modeled by a gamma distribution and the model is a system of two differential equations with distributed delay, which includes the differential equations model with a discrete delay and the ordinary differential equations model as special cases. We study the stability in all three types of models. It is shown that the ODE model is globally stable while both delay models exhibit Hopf bifurcations by using the (average) delay as a bifurcation parameter. The results indicate that, differing from the cell-to-free virus spread models, the cell-to-cell spread models can produce infective oscillations in typical tissue culture parameter regimes and the latently infected cells are instrumental in sustaining the infection. Our delayed cell-to-cell models may be applicable to study other types of viral infections such as human T-cell leukaemia virus type 1 (HTLV-1). PMID- 12750835 TI - Evolutionary and dynamic stability in continuous population games. AB - Asymptotic stability under the replicator dynamics over a continuum of pure strategies is shown to crucially depend on the choice of topology over the space of mixed population strategies, namely probability measures over the real line. Thus, Strong Uninvadability, proved by Bomze (1990) to be a sufficient condition for asymptotic stability under the topology of variational distance between probability measures, implies convergence to fixation over a pure strategy x(*) only when starting from a population strategy which assigns to x(*) a probability sufficiently close to one. It does not imply convergence to x(*) when starting from a distribution of small deviations from x(*), regardless of how small these deviations are. It is, therefore, suggested that when a metric space of pure strategies is involved, another topology, hence another stability condition, may prove more relevant to the process of natural selection. Concentrating on the case of a one dimensional continuous quantitative trait, we resort to the natural Maximum Shift Topology in which an epsilon -vicinity of the fixation on a pure strategy x(*) consists of all mixed population strategies with support which includes x(*) and is in the epsilon -neighborhood of x(*). Under this topology, a relatively simple necessary and sufficient condition for replicator asymptotic stability, namely Continuous Replicator Stability (CRSS), is demonstrated. This condition is closely related to the static stability condition of Neighbor Invadability (Apaloo 1997), and slightly stronger than the condition of Continuous Stability (Eshel and Motro 1981). PMID- 12750837 TI - Cattaneo models for chemosensitive movement: numerical solution and pattern formation. AB - We derive models for chemosensitive movement based on Cattaneo's law of heat propagation with finite speed. We apply the model to pattern formation as observed in experiments with Dictyostelium discoideum, with Salmonella typhimurium and with Escherichia coli. For Salmonella typhimurium we make predictions on pattern formation which can be tested in experiments. We discuss the relations of the Cattaneo models to classical models and we develop an effective numerical scheme. PMID- 12750838 TI - Pharmacokinetics of BNP7787 and its metabolite mesna in plasma and ascites: a case report. AB - PURPOSE: BNP7787 (2',2'-dithio-bis-ethane sulfonate sodium) is a novel protector against cisplatin-induced toxicities. The pharmacokinetics of BNP7787 and its metabolite mesna were investigated in plasma and ascites of a cancer patient. We also evaluated potential pharmacokinetic interactions between BNP7787 and cisplatin. METHODS: BNP7787 and mesna were measured as mesna in deproteinized plasma and ascites using high-performance liquid chromatography with an electrochemical detector provided with a wall-jet gold electrode. RESULTS: After the i.v. administration of 41 g/m(2) BNP7787, BNP7787 and mesna had a half-life of 1.5 and 3.4 h, respectively. The AUC( infinity ) of mesna was approximately 8% of the AUC( infinity ) of BNP7787. Coadministration of cisplatin did not appear to influence the plasma concentration-time curves of BNP7787 and mesna. In ascites, approximately 0.02% of the BNP7787 dose was present as mesna, whereas approximately 4% of the dose was present as BNP7787 at the time of the maximum concentration. CONCLUSIONS: It can be concluded that the presence of ascites did not have a major impact on the pharmacokinetics of BNP7787 and coadministration of cisplatin did not influence the pharmacokinetics of BNP7787 and mesna. PMID- 12750839 TI - Phase I/II study of escalating doses of nedaplatin in combination with irinotecan for advanced non-small-cell lung cancer. AB - We conducted a phase I/II study of combination chemotherapy with nedaplatin (NDP) and irinotecan to determine the effects against advanced non-small-cell lung cancer (NSCLC) and to determine the qualitative and quantitative toxicities of the combination chemotherapy. NDP was given on day 1 and irinotecan on days 1 and 8. The treatment cycle was designed to be repeated every 3 weeks. We fixed the dose of irinotecan as 60 mg/m(2) and escalated the NDP dose from a starting dose of 50 mg/m(2) by 10-mg/m(2) increments until the maximum tolerated dose (MTD) was reached. The MTD was defined as the dose level at which at least two of three or three of six patients experienced a dose-limiting toxicity (DLT). Between April 1997 and November 2000, 42 patients were registered in the study. Of the 42 patients, 37 had no prior treatment, 3 had received whole-brain irradiation, 1 had undergone surgical resection, and 1 had had one regimen of chemotherapy before enrolling in this study. In the phase I study, we observed DLTs such as grade 4 neutropenia lasting 7 days and grade 3 diarrhea lasting 1 day in one patient at level 2, grade 3 elevated of GPT in one patient at level 3, and acute myocardial infarction in one patient at level 6. We could not determine the MTD until dose level 6 was reached, so decided on a recommended dose of 100 mg/m(2) NDP, which is recommended for NDP-alone chemotherapy. Because of prolonged neutropenia in the phase I study, we repeated the treatment every 4 weeks in the phase II study. In the phase II study, a total of 16 patients, including 6 patients from the phase I study, were registered and a total of 42 cycles were administered. Grade 3 or 4 neutropenia, grade 3 anemia and grade 3 or 4 thrombocytopenia occurred in 50%, 12% and 7% of cycles, respectively. Febrile neutropenia occurred in eight cycles (19%) but there were no severe infections. Grade 3 elevation of GPT occurred in one patient. Of the 16 patients, 7 had an objective response. Of the 42 patients, 13 achieved a partial response (PR) and the overall response rate was 31.0%. The median duration of PRs was 226 days (range 59 to 646 days). The median survival time was 341 days and the 1-year survival rate was 45.2%. In conclusion, the combination of NDP and irinotecan was highly effective and well tolerated in NSCLC. PMID- 12750840 TI - Metabolism of dexrazoxane (ICRF-187) used as a rescue agent in cancer patients treated with high-dose etoposide. AB - PURPOSE: The study was undertaken to determine the metabolism of dexrazoxane (ICRF-187) to its one-ring open hydrolysis products and its two-rings opened metal-chelating product ADR-925 in cancer patients with brain metastases treated with high-dose etoposide. In this phase I/II trial dexrazoxane was used as a rescue agent to reduce the extracerebral toxicity of etoposide. METHODS: Dexrazoxane and its one-ring open hydrolysis products were determined by HPLC and ADR-925 was determined by a fluorescence flow injection assay. RESULTS: The two one-ring open hydrolysis intermediates of dexrazoxane appeared in the plasma at low levels upon completion of dexrazoxane infusion and then rapidly decreased with half-lives of 0.6 and 2.5 h. A plasma concentration of 10 micro M ADR-925 was also detected at the completion of the dexrazoxane i.v. infusion period, indicating that dexrazoxane was rapidly metabolized in vivo. A plateau level of 30 micro M ADR-925 was maintained for 4 h and then slowly decreased. The pharmacokinetics of dexrazoxane were found to be similar to other reported data in other settings and at lower doses. CONCLUSIONS: The rapid appearance of ADR 925 in plasma may make ADR-925 available to be taken up by heart tissue and bind free iron. These results suggest that the dexrazoxane intermediates are enzymatically metabolized to ADR-925 and provide a pharmacodynamic basis for the antioxidant cardioprotective activity of dexrazoxane. PMID- 12750841 TI - Arsenic trioxide induces apoptosis in cells of MOLT-4 and its daunorubicin resistant cell line via depletion of intracellular glutathione, disruption of mitochondrial membrane potential and activation of caspase-3. AB - PURPOSE: To demonstrate that arsenic trioxide (As(2)O(3)) induces apoptosis via a mitochondrial pathway in both parent T lymphoblastoid leukemia MOLT-4 cells and cells of its daunorubicin-resistant subline, MOLT-4/DNR, expressing functional P gp. METHODS: Cell growth was measured using an MTT assay. Cell viability was determined using a dye exclusion test. Intracellular glutathione (GSH) was measured using a glutathione assay kit. Mitochondrial membrane potential (MMP) was assessed by rhodamine 123 (Rh123) staining intensity on flow cytometry. Caspase-3 activity was evaluated using a commercially available assay kit on flow cytometry. The percentage of cells undergoing apoptosis was estimated in terms of caspase(+)/PI(-) cells on flow cytometry after assessment for activation of caspase-3 by adding PI. RESULTS: MOLT-4 cells and MOLT-4/DNR cells were similarly sensitive to the apoptosis-inducing effect of As(2)O(3). Buthionine sulfoxide (BSO) and ascorbic acid (AA) rendered these cells more sensitive to As(2)O(3), whereas N-acetylcysteine (NAC) reduced this sensitivity. BSO and AA decreased, but NAC increased, the intracellular GSH contents of both MOLT-4 and MOLT-4/DNR cells. Decreasing GSH with BSO potentiated As(2)O(3)-mediated growth inhibition, disruption of MMP, activation of caspase-3 and apoptosis of cells. Clinically relevant doses of AA enhanced the anticancer effects of As(2)O(3) via the disruption of MMP, activation of caspase-3, and induction of apoptosis. In contrast, increase GSH levels with NAC attenuated all of these As(2)O(3)-mediated actions. CONCLUSIONS: The sensitivity of MOLT-4 and MOLT-4/DNR cells to As(2)O(3) was associated with the intracellular GSH content. As(2)O(3) induced apoptosis in parent MOLT-4 cells and MOLT-4/DNR cells expressing functional P-gp via depletion of intracellular GSH, and subsequent disruption of MMP and activation of caspase 3. PMID- 12750842 TI - Phase I trial of fixed dose-rate gemcitabine in combination with carboplatin in chemonaive advanced non-small-cell lung cancer: a Cancer Therapeutics Research Group study. AB - PURPOSE: To determine the maximally tolerated dose (MTD) of gemcitabine administered at a fixed dose-rate of 10 mg/m(2) per min in combination with fixed dose carboplatin, to evaluate the toxicity of this regimen and to determine the pharmacokinetics of plasma gemcitabine. METHODS: Patients with advanced stage non small-cell lung cancer (NSCLC) received carboplatin (AUC 5) on day 1 followed by gemcitabine at a fixed dose rate of 10 mg/m(2) per min in escalating durations of infusion on days 1 and 8 every 21 days. Pharmacokinetic sampling was obtained on day 1, cycle 1 of treatment. RESULTS: A total of 15 patients received carboplatin and gemcitabine in cohorts of three to six patients at three dose levels. The doses of gemcitabine studied were 600, 750, and 900 mg/m(2). The MTD was reached at 900 mg/m(2). Dose-limiting toxicities were thrombocytopenia and liver failure, and with repeated dosing neutropenia was commonly observed. The recommended phase II dose of gemcitabine was 750 mg/m(2). Partial responses were observed at 600 and 750 mg/m(2) of gemcitabine. Plasma gemcitabine did not reach steady state except in one patient with the durations of infusion studied. Plasma concentrations, however, were above 10 micro mol/l between 20 and 90 min in all patients. CONCLUSIONS: Gemcitabine administered as a 75-min infusion at a fixed dose rate of 10 mg/m(2)/min on days 1 and 8 in combination with carboplatin on day 1 every 21 days is tolerable and active in NSCLC. Pharmacokinetic studies demonstrated that the target plasma gemcitabine concentration above 10 micro mol/l was achieved. Further studies are warranted to compare this regimen against standard regimens of carboplatin and gemcitabine. PMID- 12750843 TI - Stenotrophomonas maltophilia septicemia with pyomyositis in a chemotherapy treated patient. AB - Pyomyositis is a rare complication of chemotherapy. A 35-year-old male patient with myelodysplastic syndrome developed Stenotrophomonas maltophilia bacteremia shortly after chemotherapy, and Stenotrophomonas maltophilia-related pyomyositis was encountered after recovery from neutropenia. He recovered completely after surgical drainage and a protracted course of antibiotic treatment. It is postulated that subclinical myopathy, immunosuppression secondary to the malignancy, or chemotherapeutic drugs may predispose to pyomyositis. Early recognition of this unusual complication in a cancer patient undergoing chemotherapy can prevent further catastrophes. PMID- 12750844 TI - Acute lymphoblastic leukemia with large molecular ACTH production. AB - Ectopic hormone production is very rare in hematological malignancy. Here, we describe an interesting case of acute lymphoblastic leukemia (ALL) with adrenocorticotropic hormone (ACTH) production. A 47-year-old man was admitted to our hospital with a 7-month history of hyperpigmentation. The plasma level of ACTH was markedly elevated without a circadian rhythm and the level of cortisol was normal. Examination of bone marrow aspiration revealed ALL, and no other disease as a cause of the elevated ACTH was detected. Sephadex G-75 chromatography of plasma ACTH extract revealed the existence of an abnormally large molecular ACTH (probably proopiomelanocortin) in addition to authentic 1-39 ACTH. Ectopic ACTH of low biological activity is considered to be the reason for a discrepancy in the plasma levels of ACTH and cortisol. Shortly after remission induction chemotherapy, blast cells in the peripheral blood disappeared, and the plasma level of ACTH became normal, leading to an improvement of skin pigmentation. These clinical findings and laboratory data suggested that leukemia cells in this case may produce the ACTH. PMID- 12750845 TI - Poor engraftment after allogeneic bone marrow transplantation: role of chimerism analysis in treatment and outcome. AB - We analyzed the clinical course and risk factors of 18 patients with poor engraftment after allogeneic bone marrow transplantation (BMT), defined as absolute neutrophil count below 0.1 x 10(9)/l 28 days post-BMT. Significant risks associated with non-engraftment included HLA one antigen mismatch, BMT from matched unrelated donor, and a low dose of colony-forming units-granulocyte macrophage (<10(4)/kg). Examined by a semiquantitative analysis of polymorphic microsatellite markers, donor DNA chimerism on day 28 was found to be predictive of treatment outcome. Seven patients had detectable donor DNA, varying from 43 to 100%. Five of them responded to granulocyte colony-stimulating factor (G-CSF) and achieved engraftment. Two were given further infusions of peripheral blood hematopoietic stem cells (PBSC) from the same donors, resulting in engraftment in one of them. Eleven patients had no detectable donor DNA, and none responded to G CSF. Autologous regeneration occurred in six of these patients, four after infusion of backup marrow and two spontaneously. The remaining five patients died despite the administration of PBSC from the same or different donors. Regular monitoring of donor DNA chimerism is useful in the management of patients at high risk of poor engraftment. PMID- 12750846 TI - Diaphyseal nutrient foramina in the first metatarsals in normal and hallux valgus feet: location and surgical implications. AB - We examined 47 first metatarsals from amputated lower limbs to determine the situation of the main diaphyseal nutrient foramina (NFs) in normal and hallux valgus feet. All the NFs, excepting one, were in a plantar-fibular location. The NF situation was analyzed by means of the foraminal index and three minimum distances: from NF to proximal extremity, from NF to the shaft dorsal face (NFDS) and from NF to the border of the cartilaginous coating of the metatarsal head. We found a constant location of the NF in the middle of the total metatarsal length and sexual dimorphism in NFDS (lower in females); there were no differences by side, neither by digital or metatarsal types, nor between normal and hallux valgus types. Vascular complications in some osteotomies are discussed. In the surgical design, the NF situation can be estimated from either the total or physiological metatarsal length by means of the corresponding equations as reported here. PMID- 12750847 TI - Strategy to control methicillin-resistant Staphylococcus aureus post-operative infection in orthopaedic surgery. AB - In the year 2000 the rate of infection after arthroplasty in our hospital was 9.75% and methicillin-resistant Staphylococcus aureus (MRSA) was the organism in 33% of the infected joints. In an attempt to overcome this unacceptable situation, we changed our prophylaxis regime over a period of 6 months. This involved modifying the precautionary measures for preventing surgical infections, active prophylaxis against any nasal reservoir of infection in joint implant patients, the control of health care personnel, the strict application of standard and contact precautions in all patients with MRSA, and the use of teicoplanin as prophylaxis during this 6-month period. This resulted in a definite decrease in the incidence of orthopaedic wound infections by MRSA, while the level of MRSA infection elsewhere in the hospital remained constant. Only one infection was detected during this 6-month trial, and this beneficial effect was maintained during the following 6 months. Since then, only sporadic new infections have been detected. Patients with arthroplasties performed during the study were followed for 12 months, and no new cases of MRSA infection were detected. PMID- 12750850 TI - Visualising deep lung lymphatic drainage with radioliposomes. PMID- 12750849 TI - Late-term reconstruction of lateral ankle ligaments using a split peroneus brevis tendon graft (Colville's technique) in patients with chronic lateral instability of the ankle. AB - We reviewed 14 patients with chronic lateral instability of the ankle treated by Colville's technique between 1996 and 2001. The mean patient age was 25 (20-35) years and all were men. The mean period between injury and surgery was 25 (18-32) months, and the mean follow-up was 20 (14-32) months. Twelve of the results were excellent and two were good according to the criteria of Chrisman and Snook. All patients returned to normal daily activity levels at an average of 6 months following surgery. PMID- 12750848 TI - The incorporation of different sorts of cancellous bone graft and the reaction of the host bone. A histomorphometric study in sheep. AB - We performed a morphological and histomorphometric analysis of the use of either autografts, or of frozen or freeze-dried cancellous bone allografts in sheep. A cancellous bone defect was created in the lateral portion of the distal epiphysis of the left femur. Four groups of six animals were monitored for 3 months. In the first group, the cavity was filled with autograft, in the second with frozen allograft and in the third with freeze-dried allograft. In the last group, the cavity was not filled and served as control. A study of the host bone showed that the mean trabecular width of the peripheral osteoid was greatest in the control group, while the number of osteoblasts and osteoclasts was significantly lower in the freeze-dried allograft group. However, the different bone grafts that were used to fill the cavity showed a greater trabecular width and area in the autografts. Among the frozen allografts, these measurements were also greater than in the freeze-dried allograft group. The "erosion surface" of the freeze dried allograft group was also found to be three times greater, and there were a larger number of osteoclasts and osteoclastic nuclei. We concluded that the "lyophilised" allografts were re-absorbed rapidly and that there were no major morphological differences between the frozen allografts and the autograft groups. PMID- 12750851 TI - 99mTc-HMPAO and 99mTc-ECD perform differently in typically hypoperfused areas in Alzheimer's disease. AB - Technetium-99m hexamethylpropylene amine oxime (HMPAO) and (99m)Tc- N, N"-1,2 ethylene diylbis- l-cysteine diethyl ester dihydrochloride (ECD) yield significantly different images of cerebral perfusion owing to their particular pharmacokinetics. The aim of this study was to assess the topography, extension and statistical significance of these differences in Alzheimer's disease (AD). Sixty-four patients with mild to moderate AD were retrospectively selected by two European centres. Two series of patients, including 32 studied with (99m)Tc-HMPAO single-photon emission tomography (SPET) and 32 studied with (99m)Tc-ECD SPET, were matched for sex, age (+/-3 years) and severity of cognitive impairment as assessed by the Mini-Mental State Examination (MMSE) (+/-2 points), following a case-control procedure. SPET data were processed using SPM99 software (uncorrected height threshold: P=0.001). (99m)Tc-ECD SPET gave significantly higher uptake ratio values than (99m)Tc-HMPAO SPET in several symmetrical clusters, including the right and left occipital cuneus, the left occipital and parietal precuneus, and the left superior and middle temporal gyri. (99m)Tc-HMPAO SPET gave significantly higher uptake ratio values than ECD in two smaller clusters, including the hippocampus in both hemispheres. In AD, relative brain uptake of (99m)Tc-HMPAO and (99m)Tc-ECD is different in several brain regions, some of which are typically involved in AD, such as the precuneus and the hippocampus. These differences confirm the need for specific normal databases, but their impact on routine SPET reports in AD is not known and deserves an ad hoc investigation. PMID- 12750852 TI - Co-fermentation of glucose and citrate by Lactococcus lactis diacetylactis: quantification of the relative metabolic rates by isotopic analysis at natural abundance. AB - The simultaneous catabolism of citrate and glucose by growing Lactococcus lactis subsp. lactis biovar. diacetylactis to obtain energy was followed quantitatively, using a non-enrichment isotopic technique. Both citrate and glucose are precursors of pyruvate, which may either be reduced to lactate, the principle product that accumulates, or be converted to diacetyl and acetoin. Under suitable conditions, both routes regenerate NAD+. Until recently, however, the quantitative relationships between the two substrates and these three products were poorly defined. It was recently shown, by exploiting differences in natural abundance 13C/12C ratios in the two substrates, that there is no metabolic separation of the catabolism of these two carbon sources. In this study, it is shown that the relative consumption rates change throughout the growth phase, citrate being preferentially metabolised at the onset of a culture of energy depleted cells, with a subsequent evolution towards a metabolism dominated by glucose consumption. Additionally, it is shown that the relative consumption rates are influenced by environmental factors, notably initial pH and temperature. PMID- 12750853 TI - Effect of redox potential on stationary-phase xylitol fermentations using Candida tropicalis. AB - Redox potential was used to develop a stationary-phase fermentation of Candida tropicalis that resulted in non-growth conditions with a limited decline in cell viability, a xylitol yield of 0.87 g g(-1) (95% of the theoretical value), and a high maximum specific production rate (0.67 g g(-1) h(-1)). A redox potential of 100 mV was found to be optimum for xylitol production over the range 0-150 mV [correction]. A shift from ethanol to xylitol production occurred when the redox potential was reduced from 50 mV to 100 mV as cumulative ethanol (Y(ethanol)) decreased from 0.34 g g(-1) to 0.025 g g(-1) and Y(xylitol) increased from 0.15 g g(-1) to 0.87 g g(-1) (alpha=0.05). Reducing the redox potential to 150 mV did not improve the fermentation. Instead, the xylitol yield and productivity decreased to 0.63 g g(-1) and 0.58 g g(-1) h(-1) respectively and cell viability declined. The viable, stationary-phase fermentation could be used to develop a continuous fermentation process, significantly increasing volumetric productivity and reducing downstream separation costs, potentially by the use of a membrane cell-recycle reactor. PMID- 12750854 TI - Affinity selection of target cells from cell surface displayed libraries: a novel procedure using thermo-responsive magnetic nanoparticles. AB - Biotinylated thermo-responsive magnetic nanoparticles for use in affinity selection from yeast cell surface display libraries were prepared by coating magnetite nanoparticles with a thermo-responsive polymer consisting of N isopropyl acrylamide and a biotin derivative. These particles showed a reversible transition between flocculation and dispersion at around the lower critical solution temperature of 30 degrees C, above which the flocculated particles- which absorbed a large amount of avidin due to their large surface area--were quickly separable by magnet. The model library was constructed by mixing control yeast cells with target yeast cells co-displaying IgG binding protein (ZZ) and enhanced green fluorescence protein. Biotinylated IgG and avidin were subsequently added to the model library, and target cells were efficiently enriched with the biotinylated magnetic nanoparticles by avidin-biotin sandwich and ZZ-IgG interaction. The few target cells (0.001%) in the model library were enriched by up to 100% in only 5 days by an affinity selection procedure repeated four times. This novel method based on magnetic nanoparticles and a yeast cell surface display system could fulfill a wide range of applications in the analysis of protein-protein interactions and rapid isolation of novel biomolecules. PMID- 12750855 TI - Degradation of an endocrine disrupting chemical, DEHP [di-(2-ethylhexyl) phthalate], by Fusarium oxysporum f. sp. pisi cutinase. AB - The efficiency of two lypolytic enzymes (fungal cutinase, yeast esterase) in the degradation of di-(2-ethylhexyl)-phthalate (DEHP) was investigated. The DEHP degradation rate of fungal cutinase was surprisingly high, i.e. almost 70% of the initial DEHP (500 mg/l) was decomposed within 2.5 h and nearly 50% of the degraded DEHP disappeared within the initial 15 min. With the yeast esterase, despite the same concentration, more than 85% of the DEHP remained even after 3 days of treatment. During the enzymatic degradation of DEHP, several DEHP-derived compounds were detected and time-course changes in composition were also monitored. During degradation with fungal cutinase, most DEHP was converted into 1,3-isobenzofurandione (IBF) by diester hydrolysis. In the degradation by yeast esterase, two organic chemicals were produced from DEHP: IBF and an unidentified compound (X). The final chemical composition after 3 days was significantly dependent on the enzyme used. Fungal cutinase produced IBF as a major degradation compound. However, in the DEHP degradation by yeast esterase, compound X was produced in abundance in addition to IBF. The toxic effects of the final degradation products were investigated, using various recombinant bioluminescent bacteria and, as a result, the degradation products from yeast esterase were shown to contain a toxic hazard, causing oxidative stress and damage to protein synthesis. PMID- 12750856 TI - A new labyrinthulid isolate, which solely produces n-6 docosapentaenoic acid. AB - A labyrinthulid strain, L59, was isolated from a leaf floating on seawater collected at the coastal area of Hokkaido Prefecture, Japan. Strain L59 contained only n-6 docosapentaenoic acid ( n-6 DPA) among all the long-chain polyunsaturated fatty acids. The proportion of n-6 DPA in the total fatty acids was 48.1% and the total fatty acids content in the cell dry weight was 26.6%. Many oil bodies were observed in the cell, mostly in the vicinity of cell membranes. The strain had spindle-shaped cell bodies and all cells were surrounded by ectoplasmic net elements. It was also clearly classified in the labyrinthulid group by phylogenetic analysis. In the optimum culture condition, using soybean oil and peptone as carbon and nitrogen sources, 0.53 g of n-6 DPA/l was produced at 20 degrees C in 7 days. PMID- 12750857 TI - Evolution of catabolic pathways for synthetic compounds: bacterial pathways for degradation of 2,4-dinitrotoluene and nitrobenzene. AB - The pathways for 2,4-dinitrotoluene (2,4-DNT) and nitrobenzene offer fine illustrations of how the ability to assimilate new carbon sources evolves in bacteria. Studies of the degradation pathways provide insight about two principal strategies for overcoming the metabolic block imposed by nitro- substituents on aromatic compounds. The 2,4-DNT pathway uses novel oxygenases for oxidative denitration and subsequent ring-fission. The nitrobenzene pathway links facile reduction of the nitro- substituent, a novel mutase enzyme, and a conserved operon encoding aminophenol degradation for mineralization of nitrobenzene. Molecular genetic analysis with comparative biochemistry reveals how the pathways were assembled in response to the recent appearance of the two synthetic chemicals in the biosphere. PMID- 12750859 TI - DNA sequence variation and molecular genotyping of natural killer leukocyte immunoglobulin-like receptor, LILRA3. AB - Leukocyte immunoglobulin-like receptors (LILRs) resemble killer cell immunoglobulin-like receptors (KIR) in structure and function and the KIR and LILR gene families form the major part of the leukocyte receptor cluster (LRC) of human chromosome 19q13.4. Unlike KIR, the LILR gene clusters do not vary in gene number. However, some individuals lack expression of LILRA3. This null allele has a 6.7-kb deletion, which encompasses the first six translated exons. This haplotype enabled unambiguous direct sequencing of LILRA3 alleles using genomic DNA from individuals heterozygous for the deletion. We have performed nucleotide sequencing of a 2.5-kb region within LILRA3 and identified eight bi-allelic substitutions, four of which were non-synonymous. Two from four previously identified LILRA3 cDNA sequences were confirmed and a further six alleles characterised, of which four will encode unique peptides. At least one of the polymorphic positions identified (encoding residue 84 of the first Ig domain) is likely to directly influence ligand binding. A PCR-SSP molecular genotyping system was developed and used to describe a panel of 172 Caucasoid individuals from South-East England. Six alleles were present in this group but they were unevenly distributed, as three alleles accounted for 88% of the studied chromosomes. PMID- 12750858 TI - A functional polymorphism in the promoter/enhancer region of the FOXP3/Scurfin gene associated with type 1 diabetes. AB - FOXP3/Scurfin, a member of forkhead/winged-helix proteins, is involved in the regulation of T-cell activation, and essential for normal immune homeostasis. The FOXP3/Scurfin gene is located on chromosome Xp11.23, which includes one of the type 1 diabetes susceptible loci. Therefore, we investigated whether the human FOXP3/Scurfin gene might be a new candidate gene for type 1 diabetes. We first screened the human FOXP3/Scurfin gene for microsatellite and single nucleotide polymorphisms. Next, we performed an association study between the FOXP3/Scurfin gene and type 1 diabetes. Then, the evaluation of promoter/enhancer activity of the intron with (GT)(n) polymorphism was performed by dual luciferase reporter assay. We demonstrated two regions contained microsatellite polymorphisms; one was (GT)(n), located on intron zero and the other (TC)(n) on intron 5, which were under linkage-disequilibrium. The (GT)(15) allele showed a significantly higher frequency in patients with type 1 diabetes than in controls (43.1% vs 32.6%, P=0.0027). The genotype frequencies of (GT)(15)/(GT)(15) in female patients and of (GT)(15) in male patients tended to be higher than those in female ( P=0.064) and male ( P=0.061) controls, respectively. A significant difference in the enhancer activity between (GT)(15) and (GT)(16) dinucleotide repeats was detected. In conclusion, the FOXP3/Scurfin gene appears to confer a significant susceptibility to type 1 diabetes in the Japanese population. PMID- 12750861 TI - Spontaneous bilateral pneumothoraces associated with Wilms tumor metastases. PMID- 12750860 TI - Large-scale analysis of HLA peptides presented by HLA-Cw4. AB - A large number of HLA-Cw4 (Cw *0402) peptides were purified, sequenced, and identified from breast and ovarian carcinoma cell lines. HLA-Cw4 molecules were expressed in these cells as soluble, secreted HLA (sHLA) and recovered from the growth medium. The peptides were separated by capillary reversed-phase HPLC and analyzed by tandem mass-spectrometry. The resulting peptides fit to some extent, but not completely, the known consensus of the Cw4 peptide-binding motif. Among the identified peptides, there are a few that originate from proteins of possible interest for cancer immunotherapy or diagnostics, including mucin-5B, ART-1, fatty acid synthase, putative prostate cancer tumor suppressor, DNA topoisomerase 1, and Rac1. This work demonstrates that large-scale identification of HLA peptides recovered from sHLA is an advantageous approach for establishing the HLA peptide consensus of different haplotypes and the identification of useful peptides for treatment of diseases such as cancer, viral, and autoimmune diseases. PMID- 12750862 TI - Ultrasound screening for developmental dysplasia of the hip. AB - BACKGROUND: Clinical examination of newborns has been shown to be inadequate for the early detection of developmental dysplasia of the hip (DDH). It is debatable whether US examination is a valid alternative. OBJECTIVE: To contribute further knowledge to the natural history of DDH; to examine the distribution of hip morphology as classified by Graf according to sex and risk factors in an unselected Italian population; to propose a temporal pattern of US screening of all newborns to detect DDH. MATERIALS AND METHODS: All newborns (n=8,896) sequentially delivered in the Maternal and Child Health Hospital of Milan underwent US examination in the first week of life and, when findings were within normal limits, in the third month of life. Subjects categorised at birth as Graf type 2a with alpha angle between 50 degrees and 52 degrees, underwent a further US examination at the end of the first month of life. Subjects with ambiguous findings at the 3-month examination were re-examined at the end of the fourth month of life. All infants with abnormal hips abandoned the screening process and underwent treatment. RESULTS. Overall, 56 cases of DDH were identified: 34 in the first week of life examination, 10 at 1 month; 10 at 3 months and 2 at 4 months. CONCLUSIONS. A two-step US screening of newborns is recommended: at the end of the first month and within the fourth month of life. PMID- 12750863 TI - Detecting the subregion proceeding to infarction in hypoperfused cerebral tissue: a study with diffusion and perfusion weighted MRI. AB - Diffusion and perfusion weighted MRI have been widely used in ischaemic stroke. We studied 17 patients in whom ischaemic areas showed an ischaemic core, an area of infarct growth and hypoperfused but ultimately surviving tissue. Apparent diffusion coefficients (ADC) were measured on days 1, 2, and 8 in the three subregions and in contralateral control areas. Cerebral blood flow (CBF), cerebral blood volume (CBV), and mean transit time (MTT) were measured in these regions on day 1 perfusion maps. On day 1, the ischaemic core had very low ADC and CBF and increased MTT. The ADC in the ischaemic core gradually increased during the week. The area of infarct growth on day 1 had slightly but significantly decreased ADC (96% of control, P=0.028), moderately decreased CBF and increased MTT. On day 1 the hypoperfused but surviving tissue had slightly but significantly increased ADC (103% of control, P=0.001), mildly decreased CBF and increased CBV and MTT. The ADC of the area of infarct growth decreased to the same level as in the ischaemic core on days 2 and 8. That of surviving tissue was still above normal on day 2 (103% of control), but had returned to the normal level by day 8. Measurement of ADC combined with perfusion MRI may help distinguish different subregions in acutely hypoperfused brain. PMID- 12750864 TI - Contrast enhancement in Lhermitte-Duclos disease of the cerebellum: correlation of imaging with neuropathology in two cases. AB - Lhermitte-Duclos disease (LDD), also known as dysplastic gangliocytoma, is a rare cerebellar lesion. It has long been regarded as avascular. We report two patients with surgically proven LDD in whom contrast enhancement was observed on MRI. Neuropathological examination revealed proliferation of veins. We suggest that peripheral enhancement of LDD probably reflects vascular proliferation of the cerebellar venous draining system, and should be considered part of the imaging features of LDD. PMID- 12750865 TI - Is there a correlation between operative results and change in ventricular volume after shunt placement? A study of 60 cases of idiopathic normal-pressure hydrocephalus. AB - In patients with communicating or normal-pressure hydrocephalus, ventricular volume decreases following implantation of differential pressure valves. We implanted hydrostatic (Miethke dual-switch) valves in 60 patients with normal pressure hydrocephalus (NPH) between September 1997 and December 2001. The patients underwent CT 1 year after operation, and we measured the Evans index. Although 83% of the patients showed no change in ventricular volume as assessed by this index, 72% nevertheless showed good to excellent and 16% satisfactory clinical improvement, while 12% showed no improvement. Moderate or marked reduction in ventricular size was observed in 17%, of whom 40% of these patients showed good to excellent and 20% satisfactory clinical improvement; 40% showed unsatisfactory improvement. The favourable outcome following implantation of a hydrostatic shunt thus did not correlate with decreased ventricular volume 1 year after operation, better outcomes being observed in patients with little or no alteration in ventricular size than in those with a marked decrease. Postoperative change in ventricular volume in NPH thus does not have the same significance as in patients with high-pressure hydrocephalus. PMID- 12750866 TI - Intracerebral hemorrhage associated with Sneddon's syndrome: is ischemia-related angiogenesis the cause? Case report and review of the literature. AB - Sneddon's syndrome is characterized by livedo reticularis and multiple ischemic infarcts often associated with antiphospholipid antibodies. Intracerebral hemorrhage (ICH) is unusual in Sneddon's syndrome and has not been reported as the presenting complaint. We report a 38-year-old woman with a history of two miscarriages, Raynaud's phenomenon and livedo reticularis who presented acutely with ICH. Angiography showed prominent leptomeningeal and transdural anastomoses (pseudoangiomatosis). Anticardiolipin antibodies were positive. A right frontal brain biopsy failed to reveal vasculitis and a skin biopsy was nonspecific. MRI showed residual intracerebral hemorrhage (ICH), diffuse atrophy, multiple small white matter infarcts and leptomeningeal enhancement. This is the first report of Sneddon's syndrome presenting with an ICH. It shares features with the Divry-van Bogaert syndrome. We discuss the cause of the pseudoangiomatosis pattern and its role in the genesis of the hemorrhage and suggest that cerebral angiography should be done in every patient with Sneddon's syndrome, as it could impact therapy. PMID- 12750867 TI - Application of solid-phase microextraction for the determination of pyrethroid residues in vegetable samples by GC-MS. AB - A solid-phase microextraction (SPME) method has been developed for the determination of 7 pyrethroid insecticides (bifenthrin, lambda-cyhalothrin, permethrin, cyfluthrin, cypermethrin, fenvalerate, and tau-fluvalinate) in water, vegetable (tomato), and fruit (strawberry) samples, based on direct immersion mode and subsequent desorption into the injection port of a GC/MS. The SPME procedure showed linear behavior in the range tested (0.5-50 microg L(-1) in water and 0.01-0.1 mg kg(-1) in tomato) with r(2) values ranging between 0.97 and 0.99. For water samples limits of detection ranged between 0.1 and 2 microg L(-1 )with relative standard deviations lower than 20%. Detection limits for tomato samples were between 0.003 and 0.025 mg kg(-1) with relative standard deviations around 25%. Finally, the SPME procedure has been applied to vegetable (tomato) and fruit (strawberry) samples obtained from an experimental plot treated with lambda-cyhalothrin, and in both cases the analyte was detected and quantified using a calibration curve prepared using blank matrix. SPME has been shown to be a simple extraction technique which has a number of advantages such as solvent free extraction, simplicity, and compatibility with chromatographic analytical systems. Difficulties with the correct quantification in a complex matrix are also discussed. PMID- 12750869 TI - Determination of titanium by slurry sampling graphite furnace atomic absorption spectrometry with the use of fluoride modifiers. AB - A method for the determination of titanium in graphite furnace atomic absorption spectrometry with slurry sampling was developed. Titanium forms thermally stable carbides in the graphite tube that leads to decreased sensitivity and severe memory effects. Various fluorinating agents, BaF(2), NH(4)F, and CHF(3) (Freon 23) were therefore examined in order to reduce or eliminate these problems. Ti was determined, at various concentration levels, in certified reference materials (CRMs) using ultrasonic slurry sampling graphite furnace atomic absorption spectrometry (USS-GFAAS). The three CRMs, GBW 07601 (Human Hair Powder), GBW 07602 (Bush Branches and Leaves), and GBW 07411 (Chinese Soil), contained 2.7 microg g(-1), 95 microg g(-1), and 0.41% Ti, respectively. For comparison, determinations of Ti were made with modifiers (BaF(2) and NH(4)F) and without modifier, using 5% CHF(3) (in argon) for cleaning the graphite furnace. Good accuracy was obtained using aqueous Ti standards for calibration. A homogeneity study showed that Ti was evenly distributed in all the samples at the mg-microg level. The relative standard deviations (RSDs) obtained for the three CRMs were 16%, 11%, and 8% ( n=30). In spite of the wide range of Ti concentrations in the present samples, the same wavelength (365.4 nm) could be used for analysis by varying the slurry sample concentration. The precision was best for the material with the highest titanium content in spite of the fact that only 3 microg of sample was introduced into the furnace. PMID- 12750868 TI - Potential parameters for the detection of hGH doping. AB - The aim of our hGH application study with non-competitive athletes was the investigation of selected serum parameters from different processes affected by hGH. Fifteen athletes (age 21-33, mean 24) were treated with 0.06 IU hGH/kg BW per day or placebo (10 hGH, 5 placebo) respectively for 14 days. Blood samples were taken prior to, during and until 10 weeks after treatment. The concentrations of the following markers were determined in relevant serum samples: IGF-I, IGFBP-3, ALS, PIIINP, PINP, osteocalcin, and leptin. The IGF-I concentration increased rapidly within the hGH treatment group and showed significantly higher levels compared to baseline even 3 days after application. The response of the IGFBP-3 to the hGH applications was lower in comparison to IGF-I. The hGH group showed an increasing IGFBP-3 compared to baseline from day 4 till day 15. The response of PIIINP to hGH is clearly delayed compared to the IGF I axis, but the PIIINP concentration remains on an increased level for a longer period (from day 4 until day 21). The time course and the extent of response varied strongly interindividually. PINP and osteocalcin showed only a small response to hGH applications. These parameters are characterised by a strong scattering of base values compared with the small response. In the hGH treatment group very different leptin concentrations were found at the beginning of the study, but after treatment decreasing leptin levels were observed in all cases. The determination of only one parameter will not be sufficient for detection of hGH abuse. A combination of markers by mathematical methods can be helpful to distinguish between placebo and hGH-treated athletes. By using the suggested discriminant function the data sets of hGH and placebo-treated athletes could be separated without false positive results. PMID- 12750870 TI - Preparation, homogeneity and stability studies of a candidate LRM for Se speciation. AB - A laboratory reference material (LRM) was prepared from Brazil nuts (Bertholletia excelsa) for quality control (QC) purposes of selenium speciation. The preparation of this LRM led through the usual operation steps applied during routine reference material production from biota samples-preparation of the raw material, homogenisation, storage design, checking of homogeneity, microbiological status and possible irradiation effects, and monitoring the species stability vs time at different storage temperatures. The selenium speciation studies to check species stability were carried out on a HPLC-UV-HG AFS measurement set-up. Special attention was paid to the correct identification of selenium species by applying independent HPLC separation techniques (ion pairing and anion-exchange chromatography). The concentration of selenomethionine (SeMet) and total Se content were quantified (79.9 microg g(-1) (calculated as Se) and 82.9 microg g(-1), respectively). The homogeneity and stability of this candidate reference material passed the relevant tests recommended by Bureau Communautaire de Reference (BCR). PMID- 12750871 TI - New contributions to the field of bead-injection spectroscopy-flow-injection analysis: determination of cobalt. AB - A bead-injection spectroscopy-flow-injection analysis (BIS-FIA) system with spectrophotometric detection has been developed for the determination of cobalt. A homogeneous bead suspension of Dowex 50 W resin (600 microL) previously loaded with the chromogenic reagent 1-(2-pyridylazo)-2-naphthol (PAN) was injected to fill the flow-cell. Co is injected into the carrier (pyrophosphate, pH 5) and reacts with the immobilized chromogenic reagent to form a green chelate. The analytical signal corresponds to the formation of the Co-PAN complex on the solid surface. At the end of the analysis, the beads are discarded by reversing the flow and instantaneously transported out of the system. The sensor shows both excellent selectivity, which could also be increased with a simple on-line modification to avoid interferences from Ni(II), Zn(II) and Cu(II), and good sensitivity; the detection limit was 19 ng mL(-1), the linear range 50-2000 ng mL(-1) and the RSD (%) 4.16. The method was satisfactorily applied to the determination of Co in waters, pharmaceuticals and alloy steels. PMID- 12750872 TI - Sulfonated and sulfoacylated poly(styrene-divinylbenzene) copolymers as packing materials for cation chromatography. AB - Three different types of cation exchangers were produced from four basic poly(styrene-divinylbenzene) substrates with different properties. Porous PS-DVB resin beads were functionalized by sulfonation and sulfoacylation under various conditions to produce sulfonated resins with exchange capacities of 0.03-1.80 mM g(-1). The matrix with 50% of cross-linking is most suitable for updating by the proposed technique. Sulfuric and chlorosulfonic acids were used as the reagents for sulfonation. The sulfonating conditions, capacities, and the technique of the synthesis are given. The effects of parameters of the sulfonation reaction and the composition of the reactionary mixture on ion-exchange capacity of the sorbents were investigated. Selectivity and efficiency of the separation of some inorganic cations and derivatives of amines and hydrazines on the resins obtained are compared by ion chromatography with conductometric detection. As a result, the sulfoacylated resin was proved more efficient for the separation of these analytes. The resolution of the analytes strongly depends on the degree of functionalization. The best performance about 19,000 plates m(-1) was obtained using the sulfopropionylated beads with an ion-exchange capacity of 0.3 mM g(-1). The prepared sulfoacylated cation exchanger was compared with the commercially available Dionex CS-12 packing material. It was found that the separation of alkaline ions and 1,1-dimethylhydrazine (UDMH) were much better that those for the commercial material. PMID- 12750873 TI - New colorimetric methods for the determination of trazodone HCl, famotidine, and diltiazem HCl in their pharmaceutical dosage forms. AB - Two sensitive and simple spectrophotometric methods are developed for the determination of trazodone HCl, famotidine, and diltiazem HCl in pure and pharmaceutical preparations. The methods are based on the oxidation of the cited drugs with iron(III) in acidic medium. The liberated iron(II) reacts with 1,10 phenanthroline (method A) and the ferroin complex is colorimetrically measured at 510 nm against reagent blank. Method B is based on the reaction of the liberated Fe(II) with 2,2-bipyridyl to form a stable colored complex with lambda(max )at 520 nm. Optimization of the experimental conditions was described. Beer's law was obeyed in the concentration range of 1-5, 2-12, and 12-32 microg mL(-1) for trazodone, famotidine, and diltiazem with method A, and 1-10 and 8-16 microg mL( 1) for trazodone and famotidine with method B. The apparent molar absorptivity for method A is 1.06x10(5), 2.9x10(4), 1.2x10(4) and for method B is 9.4x10(4 )and 1.6x10(4), respectively. The suggested procedures could be used for the determination of trazodone, famotidine, and diltiazem, both in pure and dosage forms without interference from common excipients. PMID- 12750874 TI - Curiosity and wonder. PMID- 12750875 TI - Native somatostatin sst2 and sst5 receptors functionally coupled to Gi/o-protein, but not to the serum response element in AtT-20 mouse tumour corticotrophs. AB - Of the five cloned somatostatin (SRIF: somatotropin release inhibitory factor) receptors (sst1-5), only sst2 and sst5 receptors appear to be endogenously expressed and functionally active in AtT-20 mouse anterior pituitary tumour cells. In this study, the presence and the functional coupling of SRIF receptors to G-protein in AtT-20 cells was evaluated by receptor autoradiography and guanosine-5'-Omicron-(3-[35S]thio)-triphosphate ([35S]GTPgammaS) binding, respectively. In addition, transcriptional effects via the serum response element (SRE) were assessed in AtT-20-SRE-luci cells, engineered to express constitutively SRE upstream of the luciferase reporter gene. [125I]LTT-SRIF-28, [125I]CGP 23996 and [125I]Tyr3-octreotide binding illustrates the high level of sst2/5 receptor in AtT-20 cell membranes. SRIF-14 and SRIF-28 produced a concentration-dependent increase in [35S]GTPgammaS binding (pEC50=6.72 and 7.45; Emax=79 and 74.9, respectively) which was completely abolished by pertussis toxin. sst2/5 receptor-selective ligands caused a concentration-dependent increase in [35S]GTPgammaS binding (pEC50=7.74-5.84; Emax=76.6-20.2) while sst1/3/4 receptor-selective ligands were devoid of activity. The binding profiles of [125I]LTT-SRIF-28 and the inhibition of cAMP accumulation correlated highly significantly with their corresponding [35S]GTPgammaS binding profiles (r=0.862 and 0.874, respectively). The effects of the sst2 receptor-preferring agonists Tyr3-octreotide and BIM 23027 on [35S]GTPgammaS binding, but not those of SRIF-14 and the sst5/1 receptor selective-agonist L-817,818, were competitively antagonised by the sst2 receptor antagonist d-Tyr8-CYN 154806 (pKB=7.36 and 7.72, respectively; slope factors not significantly different from unity). In AtT-20 SRE-luci cells, which carry a SRE-luciferase construct functioning in a very efficient manner, SRIF and its analogues did not affect luciferase activity. Taken together, these results demonstrate that in AtT-20 cells the expression of sst2 and sst5 receptors fit with their functional coupling to G(i/o)-proteins. The pharmacological implications of the existence of different ligand/receptor complexes are discussed. However, the intracellular pathways coupled to the activation of sst2 and sst5 receptors appear not to modulate the SRE-mediated transcriptional activity, suggesting that SRIF effects on gene expression coupled to mechanisms that have promoters other than SRE. PMID- 12750876 TI - Possible involvement of Ca2+-independent phospholipase A2 in protease-activated receptor-2-mediated contraction of rat urinary bladder. AB - Possible involvement of Ca(2+)-independent phospholipase A2 (iPLA2) was examined in protease-activated receptor-2 (PAR-2)-mediated contraction of the rat urinary bladder. Both PAR-2 activating peptide (PAR-2 AP; SLIGRL-NH2) and trypsin produced a concentration-dependent contractile response in the urinary bladder preparations. These contractions were significantly (p<0.01) attenuated by indomethacin (10 microM), an inhibitor of cyclooxygenase, or bromoenol lactone (BEL; 10 micro M), an inhibitor of iPLA2. On the other hand, the contractile responses to bradykinin were not significantly affected by BEL, although they were reduced by indomethacin. Arachidonyltrifluoromethyl ketone (AACOCF3; 30 microM), an inhibitor of cytosolic Ca(2+)-dependent phospholipase A2, did not affect the trypsin- and bradykinin-induced contractions. Both indomethacin and BEL had no inhibitory effect on the prostaglandin E2-induced contractions. These results suggest that PAR-2 activators and bradykinin stimulate the release of prostaglandins and thereby contract the rat urinary bladder smooth muscles. The release of prostaglandins by PAR-2 activators seems to be partly mediated by the iPLA2. PMID- 12750877 TI - Role of beta 1- and beta 2-adrenoceptors in hypertrophic and apoptotic effects of noradrenaline and adrenaline in adult rat ventricular cardiomyocytes. AB - In adult rat ventricular cardiomyocytes alpha1-adrenoceptor (AR) stimulation causes increases in protein synthesis. On the other hand beta1-AR stimulation inhibits protein synthesis, and evokes apoptotic cell death. We studied, in adult rat ventricular cardiomyocytes, effects of noradrenaline (NA), adrenaline (ADR) and phenylephrine (PE) on protein synthesis (assessed by [3H]-phenylalanine incorporation into the cardiomyocytes) in relation to effects on early apoptosis (measured by Annexin V/propidium iodide staining). PE (10(-9)-10(-5) M) induced protein synthesis was not affected by the beta1-AR blocker CGP 20712A (CGP, 300 nM) or beta2-AR blocker ICI 118,551 (ICI, 55 nM). ADR (10(-9)-10(-5) M) induced protein synthesis was enhanced by CGP and decreased by ICI. Pretreatment of the cardiomyocytes with pertussis toxin (PTX) decreased NA- and ADR- induced protein synthesis, but did not affect PE-effects. NA (10(-5) M) and ADR (10(-5) M) caused a significant increase in the number of apoptotic cells; these effects were enhanced by PTX-treatment, abolished by CGP, but not significantly affected by ICI. Furthermore, there was a significant negative correlation between catecholamine-evoked apoptosis and catecholamine-induced hypertrophic effects. We conclude that, in ventricular cardiomyocytes of adult rats, growth-promoting effects of NA and ADR are composed of alpha1A-AR mediated increase in protein synthesis and beta1-AR mediated apoptosis that counteracts increases in protein synthesis. The role of beta2-adrenoceptor appears to be a balance of antiapoptotic effects via a PTX-sensitive pathway and proapoptotic effects via a GS-adenylyl cyclase pathway. PMID- 12750878 TI - Protective effects of daviditin A against endothelial damage induced by lysophosphatidylcholine. AB - Previous investigations have indicated that endogenous inhibitors of nitric oxide synthase (NOS) such as asymmetric dimethylarginine (ADMA) may play an important role in endothelium dysfunction, and some antioxidant drugs improve endothelium function via reduction of ADMA level. The present study examined the antioxidation and endothelial protection of daviditin A, a xanthone compound. Daviditin A significantly inhibited Cu(2+)-induced low-density lipoprotein (LDL) oxidation (EC50: 38.7 microM) and scavenged 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals (EC50: 57.5 microM). Vasodilator responses to acetylcholine in rings of the isolated thoracic aorta were impaired in the presence of lysophosphatidylcholine (LPC)(5 mg/l). Daviditin A (10 or 30 microM) significantly attenuated inhibition by LPC of endothelium-dependent relaxation. Incubation of ECV304 cells with LPC (5 mg/l) for 24 h markedly elevated lactate dehydrogenase (LDH) activity and the levels of malondialdehyde (MDA) and ADMA, and decreased the content of nitric oxide (NO) and the activity of dimethylarginine dimethylaminohydrolase (DDAH). Daviditin A (1, 3 or 10 microM) significantly attenuated the increased release of LDH, increased content of MDA, and decreased level of NO induced by LPC. Daviditin A (3 or 10 microM) significantly inhibited the increased concentration of ADMA. Daviditin A (10 microM) significantly attenuated the decreased activity of DDAH. The present results suggest that daviditin A preserves endothelial dysfunction elicited by LPC, and the protective effect of daviditin A on the endothelium is related to reduction of ADMA concentration. PMID- 12750880 TI - Comment on: "Safety of percutaneous dilational tracheostomy in patients ventilated with high positive end-expiratory pressure". PMID- 12750879 TI - Three-year effectiveness of intravenous pamidronate versus pamidronate plus slow release sodium fluoride for postmenopausal osteoporosis. AB - All currently available and approved therapies for osteoporosis inhibit bone resorption. But, despite their great value, antiresorptive agents are generally not associated with dramatic increases in bone mass. In light of these data, the aim of our prospective, placebo-controlled, randomized clinical trial, with a 3 year follow up, was to examine the effects of cyclic intravenous pamidronate and fluoride in combination, versus pamidronate alone, on bone mineral density (BMD) at vertebral and femoral levels, biochemical markers of bone turnover, IGF-1 serum levels, and safety and tolerability in 40 postmenopausal women with osteoporosis. During the treatment period, pamidronate alone reduced both markers of bone formation and bone resorption, resulting in an increase of BMD, after 3 years, of 7.07% at the lumbar level and of 6.76% at the femoral level. In the group treated with pamidronate and fluoride, markers of bone turnover had a different trend: after 3 years, there was a lower reduction of bone resorption and an increase of bone formation markers, with a concomitant increase in IGF-1 levels. This resulted, after 3 years of treatment, in a marked variation of BMD at the lumbar level (+12.74%) and a reduced, but still significant, increase at the femoral level (3.89%). Spine radiography and clinical evaluation did not reveal any vertebral fractures in either treatment group. In conclusion, the combined use of pamidronate and fluoride produced somewhat larger, continuous increases in BMD, at the lumbar level, than pamidronate alone. PMID- 12750882 TI - In vitro evaluation of aerosol bronchodilator delivery during mechanical ventilation: pressure-control vs. volume control ventilation. AB - OBJECTIVE: To determine the effect of inspiratory time and inspiratory flow pattern on albuterol delivery by aerosol during mechanical ventilation. DESIGN: A ventilator was connected to a lung model and set to deliver a tidal volume of 0.6 l, PEEP 5 cmH(2)O, and respiratory rate of 15/min. We evaluated inspiratory times of 1 and 2 s, lung mechanics of 0.05 l/cmH(2)O compliance and 50 cmH(2)O/l/s resistance, or 0.02 l/cmH(2)O compliance and 5 cmH(2)O/l/s resistance, and three inspiratory flow patterns (constant flow volume-controlled ventilation, descending ramp flow volume-controlled ventilation, and pressure-controlled ventilation). Albuterol was delivered into the ventilator circuit by a nebulizer containing 5 mg (4 ml) albuterol or a pMDI and spacer (four actuations; 360 micro g). A filter between the Y-piece and the lung model collected the aerosol, which was analyzed for albuterol by spectrophotometry. RESULTS: For the nebulizer there were significant differences in albuterol delivered for inspiratory time, flow pattern, and lung mechanics. For the pMDI there were no significant differences for the amount of albuterol delivered for inspiratory time, flow pattern, or lung mechanics. CONCLUSIONS: Albuterol delivery by nebulizer is affected by inspiratory time and inspiratory flow pattern. When a pMDI is used, the amount of albuterol delivered is not affected by inspiratory flow pattern or inspiratory time. PMID- 12750883 TI - Chlamydia trachomatis-infected macrophages induce apoptosis of activated T cells by secretion of tumor necrosis factor-alpha in vitro. AB - Chlamydia trachomatis-infected macrophages induce T cell apoptosis. This ability might promote intracellular survival of Chlamydia and perpetuate chronic chlamydial infection. The purpose of this study was to examine the molecular mechanisms by which C. trachomatis-infected macrophages induce T cell apoptosis. Monocytes and T cells were isolated from the peripheral blood of healthy donors. Macrophages were infected with C. trachomatis, and autologous T cells were stimulated by mitogen. After 6 days, both populations were cultured together using a two-chamber transwell membrane system to differentiate between mechanisms involving either cell-to-cell contact or secretion of apoptotic factors. Apoptotic T cells were identified by propidium iodide through-flow cytometry, and tumor necrosis factor-alpha (TNF-alpha) concentrations were measured by enzyme linked immunosorbent assay. Antagonists of TNF-alpha, the Fas (CD95) molecule, TNF-related apoptosis-inducing ligand (TRAIL), and catalase were added to differentiate between the pathways of apoptosis. C. trachomatis-infected macrophages significantly induced T cell apoptosis by cell-to-cell contact (mean +/- standard deviation, 30+/-4%; P<0.001) and by humoral mechanisms (mean +/- standard deviation, 22+/-3%, P<0.001). Humoral apoptosis was mediated by secretion of TNF-alpha from infected macrophages. Inhibition of secretory TNF alpha by the monoclonal anti-TNF-alpha antibody adalimumab (D2E7) blocked T cell death in vitro. In contrast, T cell apoptosis mediated by cell-to-cell contact was not inhibited by the different anti-apoptotic reagents. In summary, TNF-alpha derived from infected macrophages is an important apoptosis factor for T cell apoptosis induced by C. trachomatis-infected cells. PMID- 12750884 TI - Glomerular cell apoptosis in human lupus nephritis. AB - Disturbed apoptosis is proposed to be involved in the pathogenesis of systemic lupus erythematosus. However, the role of renal cell apoptosis in the pathogenesis and progression of human lupus nephritis is still controversial. We have investigated glomerular cell apoptosis and the clinicopathological relationship between apoptosis and immunoserological or histological findings in 22 patients with lupus nephritis using electron microscopy and the TdT-mediated dUTP-biotin nick end labeling (TUNEL) method. Resident glomerular cells as well as infiltrating neutrophils undergoing apoptosis were observed in 12 of 20 patients with lupus nephritis using electron microscopy. TUNEL-positive cells were recognized in 93% of patients with diffuse proliferative lupus nephritis (class IV) in contrast to the 20% of patients with class V. The number of TUNEL positive cells in glomeruli significantly correlated with the level of immunoserological activity of lupus, such as anti-double-stranded DNA autoantibody and consumption of plasma complement. There was a positive correlation between glomerular cell apoptosis and the degree of proliferation in lupus nephritis. These data suggest that apoptosis is increased, but not decreased in glomeruli from patients with lupus nephritis. The signals that could induce glomerular cell apoptosis in lupus nephritis will need to be identified. PMID- 12750885 TI - Endoreduplication in conjunction with tumor progression in an aneuploid laryngeal squamous cell carcinoma. AB - We report the case of a 58-year-old man who presented with a squamous cell carcinoma pT1a G2 of the left vocal cord. Six months after histologically verified complete resection, the patient experienced an endolaryngeal and extralaryngeal local recurrence pT4 pN2b G2. We applied DNA flow cytometry (FCM) and comparative genomic hybridization (CGH) on both primary and recurrent tumor. The primary tumor and the endolaryngeal compartment of the relapse was an aneuploid cell clone with a FCM DNA index of 1.42 and 1.44, respectively. The extralaryngeal compartment showed a shift featuring a DNA index of 2.78. In the primary tumor and in both compartments of the recurrence there was an identical pattern of complex chromosomal imbalances as detected in CGH (CGH karyotype: rev ish enh [8q24.2-q24.3, 10q26.1-q26.3, 11q24-q25, 12q24.2-q23.33,X], dim [4q, 13q14.3-q31], amp[1p36.1-p36.2]). Hence, the recurrence was not associated with further gains and losses of chromosomal material. However, in the anterior part of the recurrence, the aneuploid tumor cell genome had completely doubled, obviously due to endoreduplication. Immunohistochemical analysis of several cell cycle regulators revealed altered expression of checkpoint proteins, pointing to a complex disturbance in cell-cycle regulation. PMID- 12750887 TI - Attentional and intentional cueing in a Simon task: an EEG-based approach. AB - Advance information about the location of a stimulus (attentional cueing) does not affect the Simon effect (a shortening of manual response times whenever the position of a stimulus that is irrelevant for the task corresponds to the side of the response). However, advance information about the side of a response (intentional cueing) enhances the Simon effect. At first sight, these well established results contradict two important assumptions about the origin of the Simon effect: (a) the effect originates at least in part in a covert shift of visual attention that forces the preparation of a response towards the location of the attentional shift and (b) interference between stimulus location and response side takes place within a response selection stage. We replicated the behavioral finding in a study that measured event-related potentials (ERPs) of the EEG. ERPs indicated that the mechanisms causing the Simon effect remain widely unaffected by advance information. Clear evidence for both response preparation and attentional shifts in the cue-target interval was found. Additionally, ERPs suggested that the increment of the Simon effect by intentional cueing might be due to perceptual factors rather than to an alteration in the mechanisms involved in the generation of a regular Simon effect. The implications of these data for the role of attention and of response selection in Simon tasks are discussed. PMID- 12750886 TI - Polycomb-group proteins are involved in silencing processes caused by a transgenic element from the murine imprinted H19/Igf2 region in Drosophila. AB - A subset of autosomal genes undergo genomic imprinting which results in expression from only the paternal or maternal chromosome. While this phenomenon is restricted to mammals and angiosperms, the underlying silencing mechanisms appear to be evolutionarily conserved. A biallelically unmethylated DNaseI hypersensitive region (A6-A4) between the imprinted Igf2 and H19 genes is conserved in humans and mice and functions as a tissue-specific maintenance element for the imprinted growth factor IGF2. In order to analyse A6-A4 for potentially conserved transcriptional maintenance properties, we have generated transgenic Drosophila harbouring the element in a reporter construct. These flies depicted silencing of the reporter genes lacZ and mini -white. The silenced state of the mini -white gene showed variegation and sensitivity to temperature changes. In addition, two members of the conserved Polycomb group, Enhancer of zeste and Posterior sex combs, were needed for repression. Polycomb group proteins are essential for gene silencing during development. Our results indicate that Polycomb group proteins may also be involved in the regulation of mammalian imprinted genes. PMID- 12750888 TI - Molecular characterization and post-transcriptional regulation of ME1, a type-I ribosome-inactivating protein from Mirabilis expansa. AB - Ribosome-inactivating proteins (RIPs) are N-glycosidases that remove a specific adenine from the sarcin/ricin (S/R) loop of the large rRNA, thus arresting protein synthesis at the translocation step. In the present study, ME1, a type-1 RIP, was cloned and sequenced from storage roots of Mirabilis expansa (Ruiz & Pavon). The full-length cDNA sequence of ME1 has 1,129 nucleotides with an open reading frame of 951 nucleotides representing 317 amino acids. Nucleotide analysis revealed that the N-terminal region of ME1 was cleaved, and the mature protein started at amino acid 34. ME1 showed very close similarities to MAP and MAP-4 from Mirabilis jalapa. Southern blot analysis revealed the presence of two homologous genes for ME1 cDNA in M. expansa. Northern blot analysis showed high levels of ME1 transcripts in primary and storage roots. Interestingly, jasmonic acid induced ME1 transcript expression in cell suspension cultures of M. expansa; however, the production of ME1 protein was not enhanced as observed by Western blot analysis. Our data suggest that ME1 has the ability to depurinate its own mRNA, thus inhibiting its translation. These observations suggest a possible mechanism by which ME1 protein levels are post-transcriptionally regulated. PMID- 12750889 TI - The sodium phosphate cotransporter family SLC34. AB - This review summarizes the characteristics of the solute carrier family SLC34 that is represented by the type ll Na/P(i)-cotransporters NaPi-lla (SLC34A1), NaPi-llb (SLC34A2) and NaPi-llc (SLC34A3). Other Na/P(i)-cotransporters are described within the SLC17 and SLC20 families. Type ll Na/P(i)-cotransporters are expressed in several tissues and play a major role in the homeostasis of inorganic phosphate. In kidney and small intestine, type ll Na/P(i) cotransporters are located at the apical sites of epithelial cells and represent the rate limiting steps for transepithelial movement of phosphate. Physiological and pathophysiological regulation of renal and small intestinal epithelial transport of phosphate occurs through alterations in the abundance of type ll Na/P(i)-cotransporters. PMID- 12750890 TI - Functional characterisation of glucose transport in bovine articular chondrocytes. AB - The adequate provision of glucose to articular chondrocytes is essential to sustain their predominantly anaerobic metabolism; glucose is also a precursor for the extracellular matrix macromolecules which these cells synthesise. Impaired glucose uptake would compromise cell function and potentially result in an imbalance of matrix synthesis and degradation, leading to osteoarthritis. We studied the glucose influx pathway into bovine articular chondrocytes using 2 deoxy- d-[(3)H]-glucose (DOG). Uptake occurs via an extracellular pH (pH(o)) insensitive, phloretin- and cytochalasin B-sensitive pathway, hallmarks of the GLUT family of facilitative glucose transporters, with a K(m) of 0.35+/-0.11 mM. Uptake was affected by a number of physiologically relevant factors: (1) raised hydrostatic pressure (1-30 MPa) inhibited DOG uptake by up to 30%; (2) interleukin-1 (IL-1beta) reduced uptake via an increase in transporter affinity; (3) glucosamine inhibited glucose uptake in a manner consistent with the actions of a competitive inhibitor. Given the involvement of IL-1beta in osteoarthritis and the protective role assigned to glucosamine, these findings implicate an important role for glucose delivery in chondrocyte energy production and matrix metabolism, which, therefore, may potentially affect the maintenance of cartilage integrity. PMID- 12750892 TI - The SLC32 transporter, a key protein for the synaptic release of inhibitory amino acids. AB - The SLC32 family comprises a single member: the vesicular inhibitory amino acid transporter (VIAAT) or vesicular GABA transporter (VGAT). It belongs to a eukaryotic-specific superfamily of H(+)-coupled amino acid transporters, which also comprises the mammalian SLC36 and SLC38 transporters. VIAAT exchanges GABA or glycine for protons. It is present on synaptic vesicles of GABAergic and glycinergic neurons, and in some endocrine cells, where it ensures the H(+) ATPase-driven uptake, and subsequent exocytotic release, of inhibitory amino acids. Despite a similar function in vesicular neurotransmitter loading, VIAAT is not related to the vesicular glutamate transporter (VGLUT, SLC17) or the vesicular monoamine transporter/vesicular acetylcholine transporter (VMAT/VACHT, SLC18) proteins. PMID- 12750891 TI - The SLC2 family of facilitated hexose and polyol transporters. AB - The SLC2 family of glucose and polyol transporters comprises 13 members, the glucose transporters (GLUT) 1-12 and the H(+)- myo-inositol cotransporter (HMIT). These proteins all contain 12 transmembrane domains with both the amino and carboxy-terminal ends located on the cytoplasmic side of the plasma membrane and a N-linked oligosaccharide side-chain located either on the first or fifth extracellular loop. Based on sequence comparison, the GLUT isoforms can be grouped into three classes: class I comprises GLUT1-4; class II, GLUT6, 8, 10, and 12 and class III, GLUT5, 7, 9, 11 and HMIT. Despite their sequence similarity and the presence of class-specific signature sequences, these transporters carry various hexoses and HMIT is a H(+)/ myo-inositol co-transporter. Furthermore, the substrate transported by some isoforms has not yet been identified. Tissue- and cell-specific expression of the well-characterized GLUT isoforms underlies their specific role in the control of whole-body glucose homeostasis. Numerous studies with transgenic or knockout mice indeed support an important role for these transporters in the control of glucose utilization, glucose storage and glucose sensing. Much remains to be learned about the transport functions of the recently discovered isoforms (GLUT6-13 and HMIT) and their physiological role in the metabolism of glucose, myo-inositol and perhaps other substrates. PMID- 12750895 TI - Elimination of response latency variability in neuronal spike trains. AB - Neuronal activity in the mammalian cortex exhibits a considerable amount of trial by-trial variability. This may be reflected by the magnitude of the activity as well as by the response latency with respect to an external event, such as the onset of a sensory stimulus, or a behavioral event. Here we present a novel nonparametric method for estimating trial-by-trial differences in response latency from neuronal spike trains. The method makes use of the dynamic rate profile for each single trial and maximizes their total pairwise correlation by appropriately shifting all trials in time. The result is a new alignment of trials that largely eliminates the variability in response latency and provides a new internal trigger that is independent of experiment time. To calibrate the method, we simulated spike trains based on stochastic point processes using a parametric model for phasic response profiles. We illustrate the method by an application to simultaneous recordings from a pair of neurons in the motor cortex of a behaving monkey. It is demonstrated how the method can be used to study the temporal relation of the neuronal response to the experiment, to investigate whether neurons share the same dynamics, and to improve spike correlation analysis. Differences between this and other previously published methods are discussed. PMID- 12750893 TI - Simultaneous segmental obstruction of bile duct and portal vein markedly changes a population of biliary and hepatic cells in human liver. AB - BACKGROUND AND AIMS: No studies have investigated histologic changes caused by simultaneous segmental obstruction of the bile duct and portal vein in human liver. PATIENTS/METHODS: Liver tissues with simultaneous obstruction of the segmental bile duct and portal vein (O(+/+) liver), with segmental bile duct obstruction alone (O(+/-) liver), and without obstruction (O(-/-) liver) were obtained from patients who underwent hepatectomy, and studied morphologically and immunohistochemically. RESULTS: In O(+/+) liver, the proportional area consisting of hepatocytes was significantly less (31.0+/-25.8%) than in O(+/-) liver (78.4+/ 18.9%) or O(-/-) liver (86.5+/-9.2%). In contrast, the proportional area consisting of biliary epithelial cells was significantly higher in O(+/+) liver (9.1+/-6.1%) than in O(+/-) liver (1.6+/-1.5%) or O(-/-) liver (0.7+/-0.6%). The proportional area consisting of fibrous tissue also was significantly higher in O(+/+) liver than in the other two groups. In O(+/+) liver, some cells located at the periphery of hepatocyte areas were immunoreactive for both hepatocyte and biliary epithelial cell markers. CONCLUSION: Simultaneous segmental obstruction of the bile duct and portal vein induces a marked ductular increase, periportal fibrosis, and a reduction in the number of hepatocytes in human liver tissue. PMID- 12750896 TI - Effect of cross-trial nonstationarity on joint-spike events. AB - Common to most correlation analysis techniques for neuronal spiking activity are assumptions of stationarity with respect to various parameters. However, experimental data may fail to be compatible with these assumptions. This failure can lead to falsely assigned significant outcomes. Here we study the effect of nonstationarity of spike rate across trials in a model-based approach. Using a two-rate-state model, where rates are drawn independently for trials and neurons, we show in detail that nonstationarity across trials induces apparent covariation of spike rates identified as the generator of false positives. This finding has specific implications for the "shuffle predictor." Within the framework developed for our model, covariation of spike rates and the mechanism by which the shuffle predictor leads to wrong interpretation of the data can be discussed. Corrections for the influence of nonstationarity across trials by improvements of the predictor are presented. PMID- 12750897 TI - Analysis of higher-order neuronal interactions based on conditional inference. AB - Higher-order neural interactions, i.e., interactions that cannot be reduced to interactions between pairs of cells, have received increasing attention in the context of recent attempts to understand the cooperative dynamics in cortical neural networks. Typically, likelihood-ratio tests of log-linear models are being employed for statistical inference. The parameter estimation of these models for simultaneously recorded single-neuron spiking activities is a crucial ingredient of this approach. Extending a previous investigation of a two-neuron system, we present here the general formulation of an exact test suited for the detection of positive higher-order interactions between m neurons. This procedure does not require the estimation of any interaction parameters and additionally optimizes the test power of the statistical inference. We apply the approach to a three neuron system and show how second-order and third-order interactions can be reliably distinguished. We study the performance of the method as a function of the interaction strength. PMID- 12750898 TI - Spike synchronization and firing rate in a population of motor cortical neurons in relation to movement direction and reaction time. AB - We studied the dynamics of precise spike synchronization and rate modulation in a population of neurons recorded in monkey motor cortex during performance of a delayed multidirectional pointing task and determined their relation to behavior. We showed that at the population level neurons coherently synchronized their activity at various moments during the trial in relation to relevant task events. The comparison of the time course of the modulation of synchronous activity with that of the firing rate of the same neurons revealed a considerable difference. Indeed, when synchronous activity was highest, at the end of the preparatory period, firing rate was low, and, conversely, when the firing rate was highest, at movement onset, synchronous activity was almost absent. There was a clear tendency for synchrony to precede firing rate, suggesting that the coherent activation of cell assemblies may trigger the increase in firing rate in large groups of neurons, although it appeared that there was no simple parallel shifting in time of these two activity measures. Interestingly, there was a systematic relationship between the amount of significant synchronous activity within the population of neurons and movement direction at the end of the preparatory period. Furthermore, about 400 ms later, at movement onset, the mean firing rate of the same population was also significantly tuned to movement direction, having roughly the same preferred direction as synchronous activity. Finally, reaction time measurements revealed a directional preference of the monkey with, once again, the same preferred direction as synchronous activity and firing rate. These results lead us to speculate that synchronous activity and firing rate are cooperative neuronal processes and that the directional matching of our three measures--firing rate, synchronicity, and reaction times--might be an effect of behaviorally induced network cooperativity acquired during learning. PMID- 12750899 TI - Early and late patterns of stimulus-related activity in auditory cortex of trained animals. AB - Epidural electrocorticograms over the right auditory cortex (field AI) were measured using implanted 18-channel (3 x 6) electrode arrays in four animals (Mongolian gerbil) trained to discriminate between a rising and a falling frequency modulated tone (frequency range 2-4 kHz). Using a previously introduced classification procedure, transient patterns of cortical activity suitable to discriminate between the rising and the falling modulation were identified. Early (locked to stimulus onset) and late (emerging at variable times poststimulus) patterns could be differentiated. Deletion of increasing numbers of randomly selected electrodes was used to determine a critical density of recording channels required to capture the discriminative power of the early and late patterns. Statistical analysis of the classification revealed a sigmoid dependence of the discriminative power from the number of remaining electrodes with an inflection point at 12 electrodes. The analysis of the minima of the classification statistic revealed that in the early patterns discriminative information was focal on regions corresponding to the tonotopic representation of the stimuli, whereas in late patterns this information seemed to be distributed nonfocally across larger cortical regions. This analysis supports the previous notion of the coexistence of topographically organized activity states related to the physical stimulus features and nontopographically organized states determined largely by intrinsic factors (Ohl et al. 2001). PMID- 12750900 TI - Models describing nonlinear interactions in graded neuron synapses. AB - An intracellular recording made from the retinal luminosity horizontal cell (LHC) demonstrated that repetitive red flashes enhanced the cell's responsiveness to red stimulus and depressed its responsiveness to green stimulus and that repetitive green flashes suppressed the cell's red response but produced little change in its green response. Based on the idea that the spectral plasticity of LHCs may reflect some synaptic efficacy changes between the LHC and various cones, a self-organizing system is proposed to investigate the possible manner of information processing and storage within the synapses. The results of model analysis suggest that the stimulus-pattern-related spectral plasticity is attributable to the excitatory interaction within the same kinds of synapses and the inhibitory interaction between different kinds of synapses. This system is able to encode and store the history of signal transmission in a graded and cumulative fashion. PMID- 12750901 TI - Time representing cortical activities: two models inspired by prefrontal persistent activity. AB - Timing information in the range of seconds is significantly correlated with our behavior. There is growing interest in the cognitive behaviors that rely on perception, comparison, or generation of timing. However, little is known about the neural mechanisms underlying such behaviors. Here we model two different neural mechanisms to represent timing information in the range of seconds. In one model, a recurrent network of bistable spiking neurons shows a quasistable state that is initiated by a brief input and typically lasts for a few to several seconds. The duration of this quasistable activity may be regarded as the neural representation of internal time obeying a psychophysical law of time recognition. Another model uses synfire chains to provide the timing information necessary for predicting the times of anticipated events. In this model, the neurons projected to by multiple synfire chains are conditioned to fire synchronously at the times when an external event (GO signal) is expected. The conditioning is accomplished by spike-timing-dependent plasticity. The two models are inspired by the prefrontal activities of the monkeys engaging in different timing-information related tasks. Thus, this cortical region may provide the timing information required for organizing various behaviors. PMID- 12750902 TI - Activity dynamics and propagation of synchronous spiking in locally connected random networks. AB - Random network models have been a popular tool for investigating cortical network dynamics. On the scale of roughly a cubic millimeter of cortex, containing about 100,000 neurons, cortical anatomy suggests a more realistic architecture. In this locally connected random network, the connection probability decreases in a Gaussian fashion with the distance between neurons. Here we present three main results from a simulation study of the activity dynamics in such networks. First, for a broad range of parameters these dynamics exhibit a stationary state of asynchronous network activity with irregular single-neuron spiking. This state can be used as a realistic model of ongoing network activity. Parametric dependence of this state and the nature of the network dynamics in other regimes are described. Second, a synchronous excitatory stimulus to a fraction of the neurons results in a strong activity response that easily dominates the network dynamics. And third, due to that activity response an embedding of a divergent convergent feed-forward subnetwork (as in synfire chains) does not naturally lead to a stable propagation of synchronous activity in the subnetwork; this is in contrast to our earlier findings in isolated subnetworks of that type. Possible mechanisms for stabilizing the interplay of volleys of synchronous spikes and network dynamics by specific learning rules or generalizations of the subnetworks are discussed. PMID- 12750903 TI - Internal models for visual perception. AB - Although the extrapolation of past perceptual history into the immediate and distant future is a fundamental phenomenon in everyday life, the underlying processing mechanisms are not well understood. A network model consisting of interacting excitatory and inhibitory cell populations coding for stimulus position is used to study the neuronal population response to a continuously moving stimulus. An adaptation mechanism is proposed that offers the possibility to control and modulate motion-induced extrapolation without changing the spatial interaction structure within the network. Using an occluder paradigm, functional advantages of an internally generated model of a moving stimulus are discussed. It is shown that the integration of such a model in processing leads to a faster and more reliable recognition of the input stream and allows for object permanence following occlusion. The modeling results are discussed in relation to recent experimental findings that show motion-induced extrapolation. PMID- 12750904 TI - Exposure to carcinogenic polycyclic aromatic compounds and health risk assessment for diesel-exhaust exposed workers. AB - OBJECTIVES: Workers' exposure to diesel exhaust in a bus depot, a truck repair workshop and an underground tunnel was determined by the measuring of elemental carbon (EC) and 15 carcinogenic polycyclic aromatic compounds (PACs) proposed by the US Department of Health and Human Services/National Toxicology Program (NTP). Based on these concentration data, the genotoxic PAC contribution to the diesel exhaust particle (DEP) lung-cancer risk was calculated. METHOD: Respirable particulate matter was collected during the summer and winter of 2001 (except for in the underground situation) and analysed by coulometry for EC and by GC-MS methods for PACs. The use of potency equivalence factors (PEFs) allowed the studied PAC concentrations to be expressed as benzo[a]pyrene equivalents (B[a]P(eq)). We then calculated the lung-cancer risk due to PACs and DEPs by multiplying the B[a]P(eq) and EC concentrations by the corresponding unit risk factor. The ratio of these two risks values has been considered as an estimate of the genotoxic contribution to the DEP cancer risk. RESULTS: For the bus depot and truck repair workshop, exposure to EC and PACs has been shown to increase by three to six times and ten times, respectively, during winter compared to summer. This increase has been attributed mainly to a decrease in ventilation during the cold. With the PEF approach, the B[a]P(eq) concentration is five-times higher than if only benzo[ a]pyrene (B[a]P) is considered. Dibenzopyrenes contribute an important part to this increase. A simple calculation based on unit risk factors indicates that the studied PAC contribution to the total lung-cancer risk attributed to DEPs is in the range of 3-13%. CONCLUSIONS: The 15 NTP PACs represent a small but non-negligible part of lung-cancer risk with regard to diesel exposure. From this point of view, the dibenzopyrene family are important compounds to be considered. PMID- 12750905 TI - Immunolocalization of vacuolar system-associated protein-60 (VASAP-60). AB - We have characterized the localization of the protein termed VASAP-60 in different bovine tissues and cell lines, and have investigated if VASAP-60 interacts with other proteins. Monospecific polyclonal antibodies were raised against distinct fragments of VASAP-60: NH(2) (V(22) to Q(234)), central (A(246) to S(418)), and COOH (L(416) to L(533)). These three antibodies recognized an 88 kDa protein in immunoblotting analysis. The calculated Mr of VASAP-60 derived from its cDNA (60.1 kDa) was significantly lower than its Mr estimated by SDS PAGE, and this was mainly attributed to the glutamic acid- and aspartic acid-rich composition of its central region (A(246) to S(418)). A 58-kDa proteolytically processed form of VASAP-60 was also identified. Immunocytochemistry demonstrated that VASAP-60 is found predominantly in the perinuclear region, colocalized with calnexin in the endoplasmic reticulum (ER), and partially colocalized with the endocytic marker DAMP. Immunohistochemical localization of VASAP-60 also demonstrated its presence within specialized vesicular structures not related to the ER. Immunoprecipitation using extracts prepared from S(35)Met/Cys metabolically labeled cells demonstrates that VASAP-60 interacts with 116-, 48.5 , and 26.5-kDa proteins. Therefore, VASAP-60 was found to be more widely distributed in the vacuolar system than anticipated, suggesting that VASAP-60 may function in intracellular transport events, rather than being an exclusive component of the quality control mechanism of newly synthesized proteins as thought previously. PMID- 12750906 TI - ML-7 inhibits exocytosis of superoxide-producing intracellular compartments in human neutrophils stimulated with phorbol myristate acetate in a myosin light chain kinase-independent manner. AB - ML-7, (5-iodonaphthalene-1-sulfonyl) homopiperazine, is commonly employed as a myosin light chain kinase (MLCK) inhibitor. In the present study, we demonstrated that ML-7 affects the superoxide (O(2)(-))-producing system of human neutrophils in an MLCK-independent manner. Human neutrophils were stimulated with phorbol myristate acetate (PMA), which does not activate MLCK. ML-7 inhibited extracellular release, but not intracellular production of O(2)(-) in the stimulated cells. Fluorescence microscopy revealed the generation of O(2)(-) at intracellular compartments in the stimulated cells exposed to ML-7. At the electron microscopic level, the reaction product of NADPH oxidase activity was found in intracellular compartments. ML-7 strongly inhibited the association of the oxidant-producing intracellular compartments with the plasma membrane. Furthermore, the upregulation of alkaline phosphatase activity, a marker enzyme of the oxidant-producing intracellular compartments, was also inhibited by ML-7. These findings indicate that ML-7 inhibits the fusion of the oxidant-producing intracellular compartments to the plasma membrane resulting in the inhibition of the extracellular release of O(2)(-) in PMA-stimulated human neutrophils in an MLCK-independent manner. PMID- 12750908 TI - The role of dendritic cells in the pharynx. PMID- 12750907 TI - Detection of RNA viruses in sudden infant death (SID). AB - Continuing previous work, reverse transcriptase polymerase chain reaction approaches for the detection of RNA viruses (influenza A+B, parainfluenza virus 3, respiratory syncytial virus) were developed. A total of 118 infant deaths, including 13 cases of non-natural death (nND), 78 cases of natural death (ND) without morphological signs of interstitial pneumonia (IP), 27 cases showing IP (ND+IP), were investigated using frozen lung tissue ( N=100) and paraffin embedded material ( N=18). In five of the autopsy cases (ND+IP) the influenza B virus genome could be detected and the other types of viruses were completely negative. Together with previous results (detection of adenoviruses and cytomegaloviruses) in the same groups, the frequency of virus detection in the cases with IP was 48% compared with 14% in the ND without IP and 7% in the nND. Significant differences in the frequency of virus detection were also obtained when the cases were divided in SIDS and non-SIDS. The results obtained indicate an association between IP and some viruses and support the hypothesis that respiratory virus infections could act as trigger in sudden infant death. PMID- 12750909 TI - Idiopathic subglottic stenosis: management by endoscopic and open-neck surgery in a series of 30 patients. AB - We describe our experience in the diagnosis and treatment of idiopathic subglottic stenosis (ISS), a rare pathological entity involving the subglottic larynx and the first tracheal rings and affecting virtually only females. Its diagnosis can be made only after the exclusion of all other known causes of subglottic stenosis. In a 17-year period, between January 1986 and June 2002, 30 patients were admitted and treated for ISS at the Department of Otolaryngology and/or the Center for Respiratory Endoscopy and Laser Therapy, the University of Brescia, Italy. Clinical, endoscopic and surgical records were retrospectively analyzed with particular emphasis on treatment (endoscopic versus open-neck procedures) and follow-up. Based on our experience, we can define endoscopic treatment by carbon dioxide or Nd:YAG laser-assisted dilatation and scar tissue resection with or without airway stenting as the treatment of choice for the initial management of ISS. After repeated endoscopic failures, open-neck surgery by laryngoplasty or laryngotracheal resection and anastomosis is strongly recommended, particularly for complex lesions longer than 1 cm. PMID- 12750911 TI - Protection from epistaxis blood aerosol contamination: a novel use of a surgical facemask. AB - Epistaxis is one of the most common otorhinolaryngological emergencies. It is also one of the most common emergencies that places medical staff at risk of blood contamination. We report a simple yet extremely effective method in which a standard surgical facemask is worn by the patient, thereby reducing blood aerosol contamination without interfering with effective management. PMID- 12750910 TI - Laryngeal chondroma presenting as an external neck mass. AB - Laryngeal chondromas are rare benign tumors originating from the cartilagenous framework of the larynx. Since its first description in 1882, about 250 cases have been reported in the literature. Depending on the size of the lesion, excision via an endoscopic or open surgical approach is the treatment of choice. The majority of these patients present with an endolaryngeal mass. To encounter an external neck mass as the reason for the initial presentation is a rare occurrence. In this article we report a case of laryngeal chondroma of cricoid cartilage origin, which presented as a neck mass. The lesion was totally excised using a transcervical approach. No tracheotomy or laryngeal stent was needed postoperatively. Despite its rarity, this diagnostic possibility should always be kept in mind for elderly patients with complaints of progressive voice changes and exertional dyspnea. PMID- 12750912 TI - Endoscopic resection of nasal haemangiopericytoma. AB - Haemangiopericytomas are vascular tumours derived from Zimmerman's pericytes; they account for about 1% of all vascular tumours. About 5% of the cases occur in the nasal cavity and usually show well-differentiated tumours with low potential for local recurrence or metastasis. The treatment of choice is surgical resection. We present a case of nasal haemangiopericytoma treated by endoscopic resection, and we emphasise the advantages of this minimally invasive method compared with traditional methods. PMID- 12750913 TI - Prognostic implications of hemoglobin levels before and after surgery as well as before and after radiochemotherapy for head and neck tumors. AB - Several studies have shown that tumor cells may develop resistance to radiotherapy, proliferating under hypoxic conditions. Following surgery, patients may develop low hemoglobin levels, which may cause low oxygen conditions. This retrospective analysis was undertaken to determine the impact of low hemoglobin levels in patients with head and neck tumors treated with combined-modality therapy (surgery and postoperative radiochemotherapy). We studied 120 patients with mostly advanced head and neck tumors (88% stage III/IV) who had undergone macroscopically complete resections of their primary tumors and lymph node metastases. At 20-277 days after surgery (median: 51.3 days), these patients received postoperative chemoradiotherapy (56.7 Gy of radiation over 28-49 days and cisplatin 6 mg/m(2) body surface area on radiation treatment days with a cumulative dose of 96 mg/m(2)). Normal hemoglobin levels were considered to be 12 g/dl for females and 13 g/dl for males. Decreased hemoglobin levels before or after surgery and before or after chemoradiotherapy were correlated with the prognosis. Preoperatively, 99 of 114 patients (87%) had normal levels of hemoglobin compared with only 20 of 107 patients (19%) postoperatively. At the onset of radiochemotherapy, the hemoglobin levels of 82 of 116 patients (71%) were within the normal range. After radiochemotherapy, however, 62 of 114 patients (54%) had normal hemoglobin levels. Univariate analysis (Kaplan-Meier method and log-rank test) showed that patients with decreased pre- or postoperative hemoglobin levels had significantly worse locoregional control ( P=0.032 and P=0.0001, respectively) and lower overall survival ( P=0.0013 and P=0.0002, respectively) than patients with normal hemoglobin levels. The 3-year locoregional control rates in patients with preoperative hemoglobin levels that were normal, were reduced by 1-2 g/dl or were reduced by more than 3 g/dl, respectively, were 78%, 55% and 50%. Correlated with normal and diminished postoperative hemoglobin levels, the 3-year locoregional control rates were 90%, 84% and 50%, respectively. There was no correlation between prognosis and hemoglobin level at the onset or after radiochemotherapy. On multivariate analysis, only the postoperative hemoglobin level remained a prognostic factor for locoregional control ( P=0.0241) and overall survival ( P=0.0080). We conclude that low postoperative hemoglobin levels resulting from blood loss may influence the efficacy of postoperative radiochemotherapy in patients with head and neck cancer. Early intervention to raise the postoperative hemoglobin level may result in better tumor control and overall survival after combined-modality therapy. PMID- 12750914 TI - Cervical metastases of microcystic adnexal carcinoma in an otherwise healthy woman. AB - Microcystic adnexal carcinoma is a rare cutaneous neoplasm characterized by slow but locally aggressive growth, which normally does not lead to systemic metastasis. Frequent local recurrences are reported, which are most likely due to insufficient operative technique. We present the fourth case of cervical ipsilateral metastatic microcystic adnexal carcinoma in an otherwise healthy woman. The patient presented with a previously diagnosed but not completely resected microcystic adnexal carcinoma in the area of the right posterior scalp and two palpable ipsilateral lymph nodes. The tumor was resected using intraoperative snap frozen histological evaluation of the resection borders. In the same procedure two lymph nodes were resected from the right neck. The lymph nodes were histologically assessed and showed infiltration by small strains of tumor cells. After exclusion of a second primary tumor, e.g., mammary carcinoma, as the cause for cervical lymph node metastases, we performed a modified radical neck dissection with resection of the sternocleidomastoid muscle and the accessory nerve, which was histologically proven to be perineurally infiltrated by tumor cells. In this second procedure the histological evaluation of the specimen showed no sign of remaining tumor infiltration. After exclusion of distant metastasis the patient was irradiated with 60 Gy. The patient is well 1 year after the initial treatment without signs of recurrence. PMID- 12750915 TI - Oropharyngeal metastasis of a Merkel cell carcinoma of the skin. AB - Merkel cell carcinoma (MCC) is an aggressive neuroendocrine tumor of the skin characterized by frequent local and regional recurrence, a high incidence of distant metastases and therefore a high mortality. Here, we report a case of an oropharyngeal metastasis of a Merkel cell carcinoma of the skin localized at the left base of the tongue causing dysphagia and the sensation of globus pharyngeus. The unusual metastatic site is presented, and diagnosis and treatment are discussed. PMID- 12750916 TI - Multilocular cervical thymic cysts in adults. A report of two cases and review of the literature. AB - Cervical thymic cysts are rare benign lesions that are hardly ever considered in the differential diagnosis of cystic neck masses. In the vast majority of cases, thymic cysts are found in infants and children. This article illustrates two cases of multilocular cervical thymic cysts in adults presenting as asymptomatic swellings in the neck. The clinical presentation, evaluation, surgical management and pathological findings are described. The possible pathogenesis as an acquired disease is reviewed and discussed. The authors recommend that, despite its rare occurrence, multilocular cervical thymic cysts should be considered in the differential diagnosis of all equivocal cases of unilateral cystic neck masses in adults. PMID- 12750917 TI - The role of dendritic cells in the pharynx. AB - The tonsils are organized as lymphoepithelial structures that play an important role in protecting both the upper respiratory and alimentary tract regions against incoming antigens. This function requires dendritic cells, professional antigen-presenting cells that act as peripheral sentinels, specializing in the uptake, processing and presentation of antigenic material. This article gives a brief review on dendritic cells with regard to their origin, life cycle and functions in the pharyngeal mucosa. The regulation of immune responses in tonsils by dendritic cells is discussed. Their importance in some disease states is also mentioned. PMID- 12750918 TI - Globus sensation and gastroesophageal reflux. AB - Recent studies suggest that gastroesophageal reflux disease (GERD) may be a major cause of globus sensation. However, the incidence and severity of GERD in patients with globus sensation without reflux symptoms are unknown. In order to establish the relationship between globus sensation in the jugular fossa and GERD, 20 patients attending our ear, nose and throat (ENT) outpatient clinic with globus sensation were investigated with 24-h pH monitoring. A four-channel pH catheter was used with the pH electrodes spaced 5 cm apart in order to detect reflux along the whole length of the esophagus. Fifteen patients complained about globus sensation only; five patients complained additionally about classical reflux symptoms. Thirteen patients showed pathologic reflux measurements. Most of the patients had reflux limited to the distal one-third of the esophagus. Patients with pathologic pH measurements were treated with proton pump inhibitors. Ten out of 13 patients improved with treatment. This study suggests that globus may be associated with reflux, and acidity does not have to reach the pharynx to produce globus sensation. PMID- 12750919 TI - A foreign body embedded in the mobile tongue masquerading as a neoplasm. AB - Foreign bodies in a mobile tongue are rarely presented to the laryngologist, because such bodies are commonly lodged superficially and are easily removed by the patients themselves or by general practitioners. Thus, it is rare that a foreign body totally embedded in the mobile part of the tongue presents as an enlarged tongue mass. We have described a 64-year-old female with a 3-month history of an enlarged mass in the anterior right tongue. Physical examination showed a mass located in the anterior right tongue, with intact mucosa and normal color. A benign tongue neoplasm was considered first. However, a fish bone totally embedded in the mobile tongue with granuloma formation was encountered during the incisional biopsy operation. Complete removal of the foreign body with granuloma was achieved under local anesthesia. There was no neuromuscular or neurosensory deficit of the tongue in the follow-up period of 2 years. Although an embedded foreign body in the mobile tongue is a rare condition, it should be considered in the work-up of a patient with an enlarged tongue mass, with or without a history of swallowing a foreign body. PMID- 12750920 TI - Spontaneous thoracic duct cyst. AB - Spontaneous and asymptomatic supraclavicular thoracic duct cysts (lymphoceles ) are rare. Only five cases have been reported so far. They are more common after surgery or trauma and have been reported in the abdomen, mediastinum, pelvis and neck. They must be differentiated from other neck cysts as failure to recognise their attachment may result in the disastrous consequence of chylothorax. A high index of suspicion is necessary, and diagnosis usually can be established by fine needle aspiration and suitable imaging. This case is reported along with a review of the literature and management options, including that of inadvertent damage to the thoracic duct. PMID- 12750921 TI - Syphilitic cervical lymphadenopathy. AB - Syphilis is well known as a great mimic, however it is not recognized as a cause of cervical lymphadenopathy. We report a case of a 21-year-old man who presented with marked unilateral cervical lymphadenopathy. He had no evidence of oropharyngeal chancres, skin or genital lesions. Computed tomography (CT) and magnetic resonance (MR) images showed multiple cervical lymphadenopathy, and serologic tests for syphilis were positive. Syphilis should still be a factor for consideration in the differential diagnosis of cervical lymphadenopathy. PMID- 12750922 TI - Hyoid myotomy with suspension under local anesthesia for obstructive sleep apnea syndrome. AB - Upper airway obstruction of patients with obstructive sleep apnea syndrome (OSAS) occurring during sleep can appear at multiple sites. Successful results of hyoid myotomy with suspension (HM) in the treatment of hypopharyngeal obstruction performed under general anesthesia have been reported. However, many OSAS patients have difficulty at the induction of anesthesia and intubation. They are predisposed to specific complications owing to anatomical abnormalities of the airway and the existence of an underlying syndrome. The purpose of this study was to assess the safety and efficacy of HM for the treatment of OSAS under local anesthesia. Thirty-two OSAS patients with hypopharyngeal obstruction underwent HM under local anesthesia. The hyoid bone was suspended to the superior aspect of the thyroid cartilage. Uvulopalatal flap was an adjunct surgical procedure for palatal obstruction. Patients had a mean age of 39.2+/-5.7 years and a body mass index of 29.3+/-2.4 kg/m(2). Data on the patients were compared from preoperative to postoperative assessment points. Statistical analysis was performed using the Student's t-test. All patients tolerated the procedure well. The mean follow-up was 8.1+/-2.4 months (range 5 to 10 months). The mean RDI decreased from 44.5+/ 8.7 to 15.2+/-5.6 ( P<0.001), and the lowest oxygen saturation increased from 82.1+/-3.7% to 87.9+/-2.7% ( P<0.01). The Epworth sleepiness scale improved from 14.1+/-2.2 to 8.2+/-2.3 ( P<0.01), and the snoring scale improved from 8.5+/-1.8 to 3.5+/-1.7 ( P<0.001). Postoperative complications included transient dysphagia in 3% of the patients and transient aspiration in 9%. Bleeding, infection and airway obstruction were not observed. Most patients had mild to moderate pain (visual analog scale 0.05). Reverse transcription-polymerase chain reaction (RT-PCR) showed that the splice variants VEGF(120 )and VEGF(164) are expressed at 3 weeks and 24 weeks, respectively. CONCLUSION: These results prove the presence of the splice variants VEGF(120 )and VEGF(164) in ruptured tendons during the healing process. The demonstrated up-regulation of VEGF in intrinsic tenocytes suggests a role for VEGF in mechanisms of angiogenesis and Achilles tendon repair. Further research is needed to evaluate if VEGF might be a possible tool to enhance the process of tendon healing. PMID- 12750927 TI - Treatment of chronic patellar dislocation with a modified Elmslie-Trillat procedure. AB - BACKGROUND: This paper describes a modification of the Elmslie-Trillat procedure that is usually performed in severe cases of habitual or recurrent patellar instability. METHODS: Eighteen knees (7 men and 8 women) treated for recurrent or habitual patellar dislocation were evaluated clinically and radiographically at a mean follow-up of 5 years (range 24 months to 9 years). The mean age at follow-up was 26.3 years (range 17-44 years). The IKDC and Kujala and Tegner scores were used for the clinical evaluation. Anteroposterior, lateral and Merchant views were done for radiographic monitoring. When the patella was still unstable during dynamic evaluation after execution of the Elmslie-Trillat procedure, the medial third of the patellar tendon was isolated and harvested with a corresponding 1 cm long and 0.5 cm wide bone plug, maintaining its insertion to the inferior medial side of the patella. This ligament was medialized and put under tension, trying to find a medial insertion that guaranteed patellar stability throughout the full range of motion. RESULTS: IKDC classified 11 knees as A (normal), 4 knees as B (almost normal), 2 knees as C (abnormal) and 1 knee as D (severely abnormal). The Kujala score showed excellent results in 16 knees, 1 fair and 1 poor knee. The mean Tegner score rose from 2 preoperatively to 5 at follow-up. The poor knee presented an over-correction of the congruence angle on radiography. On follow-up radiographs, the parameters were almost completely corrected. Statistical analysis showed a significant correction of radiograph parameters, and significantly worse results in patients who underwent trochleoplasty. CONCLUSION: The technique described tries to achieve a dynamic stability of the patella throughout the full range of motion in severe patellar instability where the Elmslie-Trillat procedure is insufficient. No recurrence of patellar instability has been observed. The stability must be obtained with dynamic control in the initial degrees of flexion, trying to avoid an excessive patellar medialization. PMID- 12750928 TI - Bilateral luxatio erecta humeri. AB - BACKGROUND: Inferior dislocation of the glenohumeral joint or luxatio erecta humeri is a rare form of injury, accounting for only 0.5% of all shoulder dislocations. Bilateral cases are even less frequent, with only eight cases reported in the literature. METHOD AND RESULT: We report one case of bilateral inferior dislocation of the shoulder. The patient made a complete functional recovery within 2 years after treatment by closed reduction and rehabilitation. CONCLUSION: Based on this case and the pertinent literature, the pathophysiology, diagnosis, and treatment of this rare injury are discussed. PMID- 12750929 TI - Ubiquitin-positive inclusions in ependymal cells. AB - Ubiquitin-positive inclusions (UbIs) have not been well studied in ependymal cells. Since we detected such UbIs in the central canals of the medulla and spinal cord while investigating UbIs in neurodegenerative diseases, we studied UbIs in the entire ependymal system of 42 patients with various neurological diseases and of 10 non-neurological controls. UbIs were located in the cytoplasm of the ependymal cells, and were round to oval in shape, measuring 4-11 microm in diameter. The UbIs were non-argyrophilic and undetectable by hematoxylin and eosin staining, but mildly reactive to periodic acid-Schiff staining with and without digestion. The UbIs were variably immunoreactive for anti-epithelial membrane antigen (EMA) antibody, but did not react with several other antibodies. The co-existence of ubiquitin and EMA was confirmed by confocal laser microscopy. Throughout the ependymal system, UbIs were variably found in ependymal cells as well as in subependymal cells. There was no significant difference in the overall incidence of either ependymal or subependymal UbIs between the patients with neurological diseases and controls. However, ependymal UbIs in the central canal were more frequent in the neurological disease patients than in controls, although there was no disease specificity. This is the first comprehensive report to show common occurrence of UbIs in the ependymal cells of adult human brains. PMID- 12750930 TI - Extrarenal retroperitoneal angiomyolipoma: management of a rare benign tumour. AB - BACKGROUND: Extrarenal retroperitoneal angiomyolipomas are benign tumours of mesenchymal tissue origin. The diagnosis of these tumours is usually based on the perioperative findings and confirmed by the resultant specimen's histology. The main differential diagnosis is liposarcoma, and accurate preoperative imaging combined with fine-needle aspiration cytology may permit a non-operative approach to these tumours. CASE PRESENTATION: We describe a single case of extrarenal retroperitoneal pelvic angiomyolipoma and provide the first case of magnetic resonance imaging of these extremely rare tumours. CONCLUSION: This article discusses the presentation, management, and outcome associated with these tumours. PMID- 12750931 TI - Neutropenic enterocolitis: is it possible to break vicious circle between neutropenia and the bowel wall inflammation by surgery? AB - BACKGROUND: Neutropenic enterocolitis is a devastating bowel wall inflammation in patients with protracted neutropenia. The approach for diagnosis and treatment is still controversial, and it is difficult and challenging to decide on what should be the next step in the management. CASE PRESENTATION: We report a 10-year-old boy who developed neutropenic enterocolitis in the course of the conservative treatment for aplastic anemia. Oral mucositis and the perianal fissure with an ulcer were important indicators for what was happening on the colonic mucosa. Colonoscopy and biopsy confirmed the diagnosis. A fast recovery was achieved with a right hemicolectomy and ileostomy. CONCLUSION: Retrospective analysis of the long-term follow-up of our patient suggests that defunctioning the colon by ileostomy breaks the vicious circle between neutropenia and bowel wall inflammation, and an early surgical intervention could be considered as an adjunctive approach to the conservative management of persistent cases. PMID- 12750932 TI - Percutaneous endoscopic gastrostomy to set up a long-term enteral feeding route in children: an encouraging result. AB - Percutaneous endoscopic gastrostomy (PEG) is now accepted as the preferred technique to establish long-term enteral feeding. A total of 30 pediatric patients (21 males and 9 females, aged from 5 months to 14 years) underwent PEG in the National Taiwan University Hospital from December 1994 to February 2001. The underlying diseases of the patients receiving PEG were neurological dysfunction ( n=26), gastrointestinal disease ( n=2) and metabolic disorders ( n=2). All patients received intravenous sedation and PEG was performed successfully. Prophylactic antibiotic treatment was given for 1 day. Tube feeding began 48 h after the placement. The Z-score of weight before and 6 months after PEG was -1.54+/-1.74 and -0.98+/-1.46, respectively, which is significant (paired t-test, p<0.0001). Accelerated weight gain was found after PEG. Complications of PEG in our patients included local wound infection ( n=15), gastrocolic fistula ( n=1), pneumoperitoneum ( n=1), dislodgement of tube ( n=6) and tube occlusion ( n=2). Of these patients 12 received button replacement after PEG and the gastrostomy tube was discontinued in 2 patients when they resumed adequate oral intake. Removal of PEG was performed in 2 patients due to complications of gastrocolic fistula and severe dislodgement. Our results encourage the use of PEG as a long-term route for nutritional supply. PMID- 12750933 TI - Groin and scrotal swellings in children aged 5 years and below: a review of 535 cases. AB - There is limited literature dedicated to the surgical problems of infants and childhood, especially diseases of the groin and scrotum, in the setting of sub Saharan Africa. We do not know the morbidity and mortality rates associated with surgical conditions of the groin and scrotum in children of our sub-region. This study was carried out among children aged 5 years and below in a regional referral hospital in Kumasi, Ghana, to determine the surgical conditions of the groin and scrotum treated at elective surgery. Inguinal hernias constituted 74% of the children with groin and scrotal diseases; this was followed by hydroceles 14.2%; then undescended testis - 10.5%; and inguinal lymphadenopathy - 0.7%. The rest were testicular abscess, seminoma and cystic lymphangioma, representing 0.2% each of groin and scrotal conditions treated in this hospital among this age group. The last three are rare conditions of the groin and scrotum as shown by our data. The recurrence rate after herniotomy was 0.7%. Orchidopexy was successfully performed in 89% of children with undescended testes; in 7.1% of cases, orchidectomy was carried out for rudimentary and small, dysplastic testes. Over 98% of the children with groin and scrotal surgical problems were treated on day-care basis. There were no recorded deaths in the series. PMID- 12750934 TI - Pre-operative ultrasonographic diagnosis of biliary atresia--with reference to the presence or absence of the extrahepatic bile duct. AB - Our aim was to evaluate the efficacy of ultrasonographic (US) examination in the pre-operative diagnosis of biliary atresia (BA) with special reference to the presence or absence of extrahepatic bile duct. Thirty consecutive neonates and infants aged 8 to 169 days (mean: 62 days) suspected of having biliary atresia were examined pre-operatively in real time B-mode ultrasonography. We used a 5 or 7.5 MHz probe of micro convex type. Patients were fasted and sedatives administered. When the common bile duct was absent, we considered it a positive finding for BA diagnosis; if not, it was considered a negative finding. A definitive diagnosis of BA was confirmed at surgery by gross morphology or intra operative cholangiography. US findings had a sensitivity of 83% (19 of 23 BA patients), a specificity of 71% (5% of 7 non BA patients) and an accuracy rate of 80%. The positive predictive value was 90% (19 of 21), while the negative predictive value was 56% (5 of 9). There were four false-negative cases. Two were BA cases with patent distal common bile duct, one was BA in which the hepatic artery was determined to be the common bile duct, and the other was a subtype of extrahepatic bile duct dilatation (the so-called, "correctable type"). We employed US criteria for visualization of the extrahepatic bile duct for pre operative diagnosis of BA. US examination referring to the presence or absence of the extrahepatic bile duct is an effective and useful method for clinical survey. PMID- 12750935 TI - Desmoplastic infantile ganglioglioma. AB - BACKGROUND: Desmoplastic infantile gangliogliomas are rare intracranial tumors that mostly occur in the first 2 years of life. They are exclusively supratentorial, generally have a voluminous size, and are partially cystic. Histologically they are characterized by a divergent astrocytic and ganglionic differentiation and a prominent desmoplastic stroma; more primitive cells may be observed, which present a higher number of mitoses and these areas can mimic the features of malignant astrocytomas. Surgery is the treatment of choice. Data available from the literature suggest that no complimentary treatment is needed in cases of complete tumor resection. Chemotherapy is an option in infants with infiltration of eloquent CNS structures and progressive disease after surgery. DIGs have generally a good prognosis: recurrence-free intervals of up to 14 years have been reported and spontaneous disappearance of tumor residuals has also been described. CASE REPORTS ND DISCUSSION:We report two cases of DIGs. The first child underwent a staged partial removal of a huge right fronto-temporo-parietal tumor when she was 2 months old. At that time histological diagnosis was anaplastic astrocytoma and on these grounds she underwent six chemotherapy cycles, with a partial reduction of the tumor residual. When she was 16 months old a new operation and complete removal of the tumor residual was performed; histological diagnosis was DIG. A review of the initial histological samples confirmed this diagnosis. Twenty-two months after surgery no tumor recurrence has been documented. This case is an example of a difficult differential diagnosis, which can lead to incorrect management choices. The second patient was operated on when he was 9 months old for a mostly cystic right temporo-parieto-occipital DIG. At surgery a deep nodule, strictly adherent to the Galen and internal cerebral veins, was not removed. MRI control 9 months after surgery showed the disappearance of the tumor residual. Eleven years after surgery no tumor recurrence has been documented. The history of this patient confirms that tumor residuals do not need complimentary treatment; indeed they do not usually grow and, as in our patient, they can spontaneously disappear. PMID- 12750936 TI - Endoscopic surgical anatomy of the paediatric third ventricle studied using virtual neuroendoscopy based on 3-D ultrasonography. AB - INTRODUCTION: Endoscopic treatment for occlusive hydrocephalus requires knowledge of individual ventricular and vascular anatomies of the ventricular system. METHODS: We studied the feasibility of virtual neuroendoscopy (VNE) based on 3-D ultrasonography (3-D US) for the identification of parenchymal and vascular anatomical landmarks of the third ventricle and its impact on the surgical planning of endoscopic third ventriculostomy (ETV) in paediatric patients. 3-D US was performed through the anterior fontanel in four infants with hydrocephalus. RESULTS: Virtual neuroendoscopy revealed the size of the foramen of Monro, anatomical landmarks of the floor of the third ventricle crucial for correct fenestration during ETV, but not the premesencephalic cistern. The basilar bifurcation was identified in relation to the floor of the third ventricle by VNE (power-Doppler ultrasonography) and confirmed intraoperatively after ETV. CONCLUSION: 3-D US-based VNE reveals detailed anatomical information on the ventricular system including the foramen of Monro and the floor of the third ventricle. Within the premesencephalic cistern vascular anatomy can be visualized, but not non-vascular structures. PMID- 12750937 TI - Influence of amplitude modulated noise on the recognition of communication signals in the grasshopper Chorthippus biguttulus. AB - The detection of acoustic communication signals in the presence of sinusoidally amplitude modulated noise was investigated in males of the grasshopper Chorthippus biguttulus. The auditory system of grasshoppers exhibits only poor spectral resolution. Hence, these animals are ideally suited to investigate noise tolerance in a system operating in the temporal domain. As a sensitive indicator for signal recognition the conspicuous phonotactic turning responses of males were recorded. The main result was that noise modulated at low frequencies (1.5-5 Hz) did not impair recognition compared to a unmodulated noise. With long stimuli even a moderate improvement of noise tolerance was observed, an effect that can probably be attributed to the existence of long troughs at low modulation frequencies during which the masking of the signal was reduced. Higher modulation frequencies (15-150 Hz), however, rendered detection and recognition increasingly difficult, due to a strong interference of the sound pulses of the masking noise with the syllable-pause structure of the species-specific signals. There are no indications for the operation of mechanisms analogous to comodulation masking release as found in vertebrates, nor for a spatial release from masking. PMID- 12750939 TI - Lack of association of tumor necrosis factor alpha gene polymorphism in patients with rheumatoid arthritis in central Taiwan. AB - The aim of this study was to investigate the association of tumor necrosis factor alpha (TNFalpha) gene promoter polymorphisms in Chinese patients with rheumatoid arthritis (RA) in central Taiwan. A total of 106 RA patients and 253 normal controls were studied. Polymerase chain reaction (PCR)-based restriction analysis was used to identify A/G polymorphism at position 308 in the promoter region of the TNFalpha, which is located at 6q21.3. For the genotype of TNFalpha-308 polymorphism, there was no statistically significant difference between RA patients and normal controls (Fisher's exact test, P=0.82). Additionally, no statistical association in the distribution of TNFalpha-308 polymorphism between rheumatoid factor (RF)-positive and -negative patients was noted. The lack of an association of TNFalpha-308 polymorphism with RA and RF in our study implies that TNFalpha-308 polymorphism cannot serve as a candidate gene marker for screening RA patients in Taiwan. PMID- 12750938 TI - Visually guided orientation in flies: case studies in computational neuroethology. AB - To understand the functioning of nervous systems and, in particular, how they control behaviour we must bridge many levels of complexity from molecules, cells and synapses to perception behaviour. Although experimental analysis is a precondition for understanding by nervous systems, it is in no way sufficient. The understanding is aided at all levels of complexity by modelling. Modelling proved to be an inevitable tool to test the experimentally established hypotheses. In this review it will by exemplified by three case studies that the appropriate level of modelling needs to be adjusted to the particular computational problems that are to be solved. (1) Specific features of the highly virtuosic pursuit behaviour of male flies can be understood on the basis of a phenomenological model that relates the visual input to the motor output. (2) The processing of retinal image motion as is experienced by freely moving animals can be understood on the basis of a model consisting of algorithmic components and components which represent a simple equivalent circuit of nerve cells. (3) Behaviourally relevant features of the reliability of encoding of visual motion information can be understood by modelling the transformation of postsynaptic potentials into sequences of spike trains. PMID- 12750941 TI - When you hear hooves, don't forget zebras along with the horses. PMID- 12750940 TI - Plasminogen activator inhibitor-1 as a link between pathological fibrinolysis and arthritis of Behcet's disease. AB - Behcet's disease (BD) commonly presents with articular manifestations and thrombotic vasculopathy. Arthritis of BD characteristically demonstrates a recurrent and nondestructive course. The pathobiological basis of the thrombotic vasculopathy and protective factors against cartilage destruction in arthritis of BD have not been elucidated. Apart from being involved in fibrinolysis and thrombolysis, the plasminogen activation system can contribute to the pathogenesis of destructive joint diseases such as rheumatoid arthritis (RA). Plasminogen activator inhibitor-1 (PAI-1) is a well known fibrinolysis inhibitor. In this study, local synovial fluid and circulating plasma PAI-1 concentrations of BD were assessed in comparison to RA patients and healthy controls to investigate the nonerosive, nondestructive nature of Behcet arthritis. Twelve patients with BD (mean age 34+/-11 years, males:females 6:6), 15 with RA (mean age 36+/-8 years, males:females 3:12), and 15 healthy adults (mean age 32+/-10 years, males:females 6:9) were included in this study. Plasma PAI-1 antigen levels and PAI-1 activities were significantly greater in BD patients than in RA patients and healthy controls ( P<0.001). Synovial fluid levels of both parameters were also higher than in RA patients ( P<0.001). These results suggest that PAI-1 may promote hypofibrinolysis of Behcet's vasculopathy and also have a protective role in the arthritis of BD. PMID- 12750942 TI - RS3PE syndrome: a clinical and immunogenetical study. AB - This study analyses the clinical, radiological, evolutive, and immunogenetical characteristics of a series of patients diagnosed with remitting seronegative symmetrical synovitis with pitting edema (RS3PE) syndrome. Reviewed were the clinical charts and human leukocyte antigen (HLA) profiles of all patients treated at a single teaching hospital fulfilling the features of this syndrome according to the definition of McCarty. Twelve cases were detected in ten men and two women aged from 62 to 85 years. Rheumatoid factor was negative in all cases, and antinuclear antibodies (ANA) were positive in two. All patients achieved complete resolution of their condition within 1 year with glucocorticoid (GC) use. Two relapsed after remission but responded again to low doses of GC. Four patients showed clinical and electrodiagnostic studies consistent with carpal tunnel syndrome. No specific HLA association could be found in this report. To date, none of these patients has developed definite rheumatic diseases, infections, or malignant diseases. Although the real nature of the syndrome is still a matter of debate, at least in our context, RS3PE remains a definite condition with an excellent prognosis. PMID- 12750943 TI - Spinach attack: a funny turn in gouty arthritis. PMID- 12750944 TI - Engineering lactic acid bacteria with pyruvate decarboxylase and alcohol dehydrogenase genes for ethanol production from Zymomonas mobilis. AB - Lactic acid bacteria are candidates for engineered production of ethanol from biomass because they are food-grade microorganisms that can, in many cases, metabolize a variety of sugars and grow under harsh conditions. In an effort to divert fermentation from production of lactic acid to ethanol, plasmids were constructed to express pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH), encoded by the pdc and adhB genes of Zymomonas mobilis, in lactic acid bacteria. Several strains were transformed with the plasmids, and transcription of pdc and adhB was confirmed by northern hybridization analysis of transformants. PDC and ADH enzyme activities were at least 5- to 10-fold lower in these bacteria compared to Escherichia coli transformed with the same plasmid. Glucose fermentations were carried out, and some, but not all, of the transformed strains produced more ethanol than the untransformed parent strains. However, lactic acid was the primary fermentation product formed by all of the transformants, indicating that ADH and PDC activities were insufficient to divert significant carbon flow towards ethanol. PMID- 12750945 TI - Expression of pigmentation genes following electroporation of albino Monascus purpureus. AB - A UV-induced albino strain of Monascus purpureus was subjected to electroporation in the presence of genomic DNA from a wild-type red strain of the fungus. Eight colonies expressed color after several weeks of growth. The growth rates of all eight color variants were significantly greater than the recipient and donor strains under some culture conditions. Spectrophotometric analysis of the pigments extracted from the color variants revealed the pigments had absorbance spectra different from the DNA donor strain. These color variants may have resulted from transformation with wild-type DNA, mutation reversion, or activation of alternative pathway(s)-i.e., new mutations-that resulted in pigment production. PMID- 12750946 TI - Role of manganese in low-pH-induced root hair formation in Lactuca sativa cv. Grand Rapids seedlings. AB - Root hair formation is induced by low pH in lettuce ( Lactuca sativa L. cv. Grand Rapids) seedlings cultured in mineral medium. The role of mineral concentrations in this phenomenon was investigated, especially for manganese. When lettuce seedlings were cultured in media that were deficient in calcium (Ca), manganese (Mn), boron (B) or molybdenum (Mo), morphological changes were induced in roots. Deficiency of other nutrients had little effect on root hair formation. Ca or B deficiency inhibited the growth of the main root and the formation of root hairs, regardless of pH. Mn or Mo deficiency increased root hair formation at pH 6 and suppressed main root growth slightly. In contrast, increasing the Mn concentration suppressed low-pH-induced root hair formation. The Mn content of roots grown at pH 4 was only about 15% of that at pH 6. In contrast, the Mo content of roots grown at low pH was about six times that of roots grown at neutral pH. These results suggest that root hair formation induced by low pH is at least partly mediated by decreased Mn uptake in root cells. PMID- 12750948 TI - Sacrococcygeal pilonidal sinus: historical review, pathological insight and surgical options. AB - Sacrococcygeal pilonidal disease is a common and well recognized entity. For many years the cause of sacrococcygeal pilonidal sinus has been matter of debate. When the treatment is considered, there was a frequent lack of success of the surgical methods of excision regarding morbidity, healing, recurrence and cure. All these factors rendered the acquired thesis of pilonidal sinus disease to be more accepted. In dealing with the pathogenesis of pilonidal sinus disease, Karydakis attributed the hair insertion process to three main factors: the invader, i.e. the loose hair; the force, which causes the insertion; and the vulnerability of the skin to the insertion of hair at the depth of the natal cleft. The sinus is initiated from a small midline opening lined by stratified squamous epithelium. Additional sinuses are frequent and have lateral openings. Malignant transformation is rare but cases of squamous cell carcinoma and verrucous carcinoma have been reported. Pilonidal sinus disease consists in a symptoms complex with presentations ranging from asymptomatic pits to painful draining lesions that are predominantly located in the sacrococcygeal region. Asymptomatic pits do not require treatment. Options for treatment of acute abscess include aspiration, drainage without curettage, and drainage with curettage. The choice of a particular surgical approach depends on the surgeon's familiarity with the procedure and perceived result in terms of low recurrence of sinus and a quick healing of resulting cavity or surgical wound. Conservative nonoperative management, closed methods, laying of track, wide excision and open drainage, wide excision and primary closure, and limited excision are the methods currently used. From the profusion of studies, it is apparent that various methods are being tried and no one method is universally acceptable. Recurrence rates vary with the technique, operator and length of follow-up. Primary closure with a lateral approach appears to give the best results. PMID- 12750947 TI - Two types of matter economy for the wintering of evergreen shrubs in regions of heavy snowfall. AB - Plant adaptation to an environment subject to heavy snowfalls was investigated in four species of evergreen shrubs growing in a Fagus crenata forest in an area of Honshu on the Sea of Japan. These shrubs stored carbohydrates in some organs before the snowy season and were covered with snow for 4-5 months. Aucuba japonica var. borealis, Camellia rusticana, and Ilex crenata var. paludosa maintained a reserve of carbohydrates during the snowy season. In Daphniphyllum macropodum var. humile, the reserve of carbohydrates decreased during winter. The respiration rates in the first three species decreased from autumn to winter, whereas the decrease in D. macropodum was slight. It was found that the first three species could use reserve carbohydrates for the growth of new shoots after the thaw, whereas in the last species the growth of new shoots depends on high photosynthetic activity in late spring. Our findings suggest some types of matter economy in evergreen shrubs for wintering in an environment of heavy snow. PMID- 12750949 TI - Experimental internal anal sphincter replacement with demucosated colonic plication. AB - BACKGROUND: Selective re-creation of a new internal anal sphincter could be indicated when the natural one is irreversibly damaged or excised. METHODS: In this preliminary experimental work, surgical techniques of internal anal sphincter replacement in pigs were investigated. After preoperative anorectal manometry, surgical procedure was done in two phases: abdominal, mobilization of the colon-rectum to the pelvic floor; and perianal, dissection of the anal canal from the external anal sphincter through the intersphincteric space. The fully mobilized anorectal segment, including the internal anal sphincter, was pulled down through the anus and resected. The distal colonic stump was then demucosated and two types of plications of the demucosated segment were accomplished, each type in three animals. The plicated segment was then returned into the anal canal, inside the external sphincter. Short-term follow-up with clinical and manometric evaluations was performed and, subsequently, histological analysis of the plicated segment, after the animals were sacrificed. RESULTS: None of the animals became incontinent. Anal manometry identified a high-pressure zone and relaxation reflex in the new anal canal. Histologic studies showed hypertrophy of smooth muscle layers without degenerative changes. CONCLUSION: This study indicates that a plication of colonic smooth muscle wall can re-create a high pressure zone in the anal canal after the internal anal sphincter has been excised. PMID- 12750950 TI - Bowel cleansing for diagnostic colonoscopy: which method is preferable? Istanbul experience. AB - BACKGROUND: A prospective randomized trial to assess the cleansing ability and tolerance of bowel preparations for colonoscopy was conducted in a group of 953 patients. METHODS: The bowel cleansing methods were: sennoside calcium, polyethylene glycol (PEG) lavage, oral sodium phosphate (NaP) in one 90-ml dose, oral NaP in 2 doses (90 ml + 45 ml), oral NaP in 2 doses (45 ml + 90 ml), oral NaP in 2 doses (45 ml + 90 ml) plus cisapride, and oral NaP in 2 doses (45 ml + 90 ml) plus domperidone. RESULTS: The cisapride-containing procedure was abandoned partially through the study because of its adverse effects. The colon cleansing ability in groups with NaP plus cisapride and NaP plus domperidone was significantly better than the other groups ( p<0.05). Tolerance to sennoside calcium and PEG lavage in comparison to the other groups was significantly worse ( p<0.05). CONCLUSION: Two doses of NaP solution (45 ml + 90 ml) plus domperidone for colon cleansing is safe, effective, rapid, inexpensive and well tolerated. PMID- 12750951 TI - Augmented unilateral gluteoplasty with fascia lata graft in fecal incontinence. AB - BACKGROUND: This study was undertaken to evaluate the results of augmented unilateral gluteoplasty with fascia lata graft in patients with fecal incontinence due to congenital or neurologic disorders. METHODS: Between January 2000 and January 2001, we performed a prospective study of 11 patients with longstanding fecal incontinence (stage C3 according to Pescatori scoring system for fecal incontinence). The patients included one girl and 10 boys aged 5-19 years who had already undergone Swenson's operation for congenital megacolon (7 patients) or abdominoperineal pull-through for high imperforate anus (4 patients). Preoperative anorectal manometric studies (maximum resting pressure, maximum squeeze pressure, rectoanal inhibitory reflex and functional anal canal length), saline enema test and magnetic resonance imaging (MRI) studies were done. Unilateral gluteoplasty augmented with fascia lata graft was wrapped around the anal canal. Biofeedback retraining was started at the beginning of the fourth postoperative week. Patients were followed both objectively and subjectively for about 6-18 months. RESULTS: Eight of 11 patients (72.7%) were clinically improved, as shown by the change in incontinence score ( p=0.01). this was confirmed by the significant changes in manometric studies, functional anal canal length, rectoanal inhibitory reflex and saline enema test. MRI done one month postoperatively showed disruption in one patient. CONCLUSIONS: Unilateral gluteoplasty augmented with fascia lata graft leads to encouraging results in patients with end-stage anal incontinence due to irreversible damage to anal sphincter with less incidence of morbidity that could be related to tension on either the muscle flap or its neurovascular bundle. PMID- 12750952 TI - Bladder and male sexual functions after autonomic nerve-sparing TME with or without lateral node dissection for rectal cancer. AB - BACKGROUND: We evaluated to what extent lateral lymph node dissection (LND) interferes with bladder and male sexual functions after radical rectal excision with adoption of careful total autonomic nerve preservation. METHODS: The study comprised 77 patients resected for mid-rectal or lower rectal cancer. Bladder and male sexual functions were studied by means of a questionnaire more than one year after surgery. Outcomes were compared between patients who received lateral LND (group 1, 65 patients) and those who did not (group 2, 12 patients). RESULTS: Only minor disturbances of bladder function were reported in 10 patients (15%) of group 1, and in 3 patients (25%) of group 2. Ten out of 37 preoperatively sexually active patients (27%) in group 1 males and one of 5 patients (20%) in group 2 males had partial or total impotency after surgery and retrograde ejaculation occurred in 3 of 27 patients (11%) and one of 4 patients (25%), respectively. Erectile impotency occurred less frequently when patients were operated during the period 1993-1996 than during 1988-1992 (11% vs. 42%, p<0.05). The age was significantly greater among patients who had loss of ejaculation. CONCLUSIONS: If lateral lymph node dissection should be used with the aim of improving radicality in rectal excision for cancer, it should be combined with careful nerve-preserving technique--which may reduce the risk of bladder and male sexual dysfunctions. PMID- 12750953 TI - The posterior sagittal trans-sphincteric and trans-rectal approaches. AB - BACKGROUND: The posterior sagittal, transphincteric approach to treat different pelvic problems has been known since the last century. Although some surgeons have embraced it and have enthusiastically advocated it's use, it has never become an overly popular technique. The purpose of this study is to evaluate the advantages and disadvantages of the approach, both from a historical perspective and from the authors' experience. METHODS: The international literature on the subject was reviewed since 1877 up to the present date. A retrospective evaluation of the authors' experience was conducted. Specific attention was paid to the final result obtained in the treatment of the original condition, surgical complications and the effect of the surgical approach. on bowel and urinary control. RESULTS: The review included 114 patients divided into two groups. Group A included 85 patients who underwent a posterior sagittal trans-sphincteric approach. Group B included 29 patients who underwent a posterior sagittal transanorectal approach, in which the anterior wall of the rectum and the sphincter were divided. Postoperative bowel control was normal in all patients except in those whose basic condition had resulted in fecal incontinence, or who had sustained an irreversible injury prior to the operation. Urinary control was normal except in cases with pre-operative incontinence. Complications included recurrence of rectogenito-urinary fistulae in 3 cases, rectocutaneous fistula in 3 patients with Hirschsprung's disease, and 2 partial wound dehisences. CONCLUSIONS: The posterior sagittal trans-sphincteric approach represents a useful technical alternative. It seems to be particularly useful to treat complications after surgery for Hirschsprung's disease, presacral masses, acquired rectogenito-urinary fistulae and idiopathic rectal prolapse. The transanorectal approach provides excellent exposure to the posterior urethra and vagina. Bowel and urinary control are not compromised. PMID- 12750954 TI - Radiofrequency ablation and plication of hemorrhoids. AB - BACKGROUND: Radiofrequency ablation is emerging as a new therapeutic method in various fields of medicine. This study describes procedure of radiofrequency ablation followed by plication of hemorrhoidal mass in advanced grades of hemorrhoids. METHODS: This non-controlled, prospective study included 300 patients (211 men) treated at Fine Morning Hospital, Laxminagar, Nagpur, India, between July 1999 and December 2000. Patients were followed over a median period of 18 months (range, 15-20 months). RESULTS: The hospital stay was less than 24 hours for all patients. After 1 week, most of the patients had symptoms like bleeding and pain. At 4 weeks, 21% complained of pruritus, but none had prolapse, incontinence or anal stenosis. At the last follow-up, 96% had relief from bleeding, while 8% of patients had developed external skin tags and 6% had asymptomatic recurrence revealed by anoscopy. CONCLUSIONS: For advanced degree of piles with prolapse as the main symptom, plication of the pile mass along with radiofrequency ablation may be used as an alternative to the various types of hemorrhoidectomies. With this treatment, hospital stay is minimized, postoperative pain in low, recurrence is low and return to work is faster. PMID- 12750955 TI - Endoscopic view of rectal amebiasis mimicking a carcinoma. AB - We report the case of a 45-year-old man with rectal amebiasis, presenting with rectal bleeding and chronic diarrhea, confirmed on rectal biopsy. The endoscopic view was highly suggestive of a carcinoma and caused confusion about its etiology. The striking difference in the endoscopic view before and after medical therapy of the tumor-like lesion was remarkable. This case illustrates the importance of an accurate histologic diagnosis before definitive treatment and highlights the mimicry of rectal carcinoma by rectal amebiasis on endoscopy. PMID- 12750956 TI - Primary retrorectal adenocarcinoma: report of a case. AB - Primary adenocarcinomas of the retrorectal (presacral) space are uncommon and usually arise from cystic lesions developing from remnants of the embryological postanal gut (tail gut cysts) containing mucous-secreting epithelium. A singular case of a patient who presented with a retrorectal mass is described. A 70-year old previously healthy man had a four-month history of perianal pain during defecation. Preoperative colonoscopy showed external compression of the lower rectum with normal mucosa; pelvic computed tomography demonstrated a 5.5 x 4.5 cm(2) retrorectal oval mass, originating from the posterior lower rectal wall, containing cystic components and some small calcifications. The tumor, after complete resection via transanal approach, was shown to be a very rare case of primary adenocarcinoma of the presacral space. The patient had an uneventful postoperative course and he is free from disease 4 years after surgery. Clinicopathological features and surgical treatment of this unusual tumor are discussed. PMID- 12750958 TI - Antimicrobial resistance and epidemiological typing of methicillin-resistant Staphylococcus aureus in Ireland (North and South), 1999. PMID- 12750957 TI - Comparison of the BDProbeTec ET system with the Cobas Amplicor PCR for direct detection of Mycobacterium tuberculosis in respiratory samples. AB - In the study presented here, the performance of the BDProbeTec ET system (Becton Dickinson, USA) was compared with the Roche Cobas Amplicor-PCR (Roche, Switzerland) to detect Mycobacterium tuberculosis complex (MTB) in clinical respiratory samples. The Bactec MGIT 960 liquid culture system (Becton Dickinson) was used as a reference method. A total of 411 samples were tested. Of the 93 culture-positive samples, both the BDProbeTec ET system and the Cobas Amplicor PCR detected 87 (sensitivity, 93.5%). When only smear-negative samples were considered, the BDProbeTec ET exhibited a sensitivity of 50% and the Cobas Amplicor-PCR 60%. Specificity was 99.7% for the BDProbeTec ET system and 100% for the Cobas Amplicor-PCR. Percent agreement between the two nucleic amplification methods was 98.7%. Inhibition occurred in three (0.7%) samples in the BDProbeTec ET system. The high sensitivity and specificity of the BDProbeTec ET system suggest it is a useful method for the rapid and direct detection of MTB in smear positive respiratory samples. PMID- 12750959 TI - Achromobacter xylosoxidans bacteremia: a 10-year analysis of 54 cases. AB - Fifty-four cases of Achromobacter xylosoxidans bacteremia diagnosed over a 10 year period in patients from 2 months to 87 years of age were reviewed. Fifty-two episodes were nosocomial. The most frequent underlying condition was neoplasm (solid or hematological). The source of infection was a contaminated intravenous catheter in 35 patients (60%) and pneumonia in 6 patients. Eight (15%) patients died. The only risk factors significantly associated with mortality were age over 65 years and neutropenia. The results of in vitro susceptibility studies of the isolates showed that antibiotic therapy with antipseudomonal penicillins or carbapenems would be a reasonable choice. An epidemiological study conducted in the hemodialysis unit showed Achromobacter xylosoxidans in tap water and on the hands of two healthcare workers but not in the hemodialysis systems. Patients were probably contaminated when healthcare workers manipulated the intravenous catheters without wearing gloves. PMID- 12750960 TI - Value of monitoring serum procalcitonin in neonates at risk of infection. PMID- 12750961 TI - Exploration of virtual mazes by rhesus monkeys (Macaca mulatta). AB - A chasm divides the huge corpus of maze studies found in the literature, with animals tested in mazes on the one side and humans tested with mazes on the other. Advances in technology and software have made possible the production and use of virtual mazes, which allow humans to navigate computerized environments and thus for humans and nonhuman animals to be tested in comparable spatial domains. In the present experiment, this comparability is extended even further by examining whether rhesus monkeys (Macaca mulatta) can learn to explore virtual mazes. Four male macaques were trained to manipulate a joystick so as to move through a virtual environment and to locate a computer-generated target. The animals succeeded in learning this task, and located the target even when it was located in novel alleys. The search pattern within the maze for these animals resembled the pattern of maze navigation observed for monkeys that were tested on more traditional two-dimensional computerized mazes. PMID- 12750962 TI - Establishment of a method of anonymization of DNA samples in genetic research. AB - As the number of the genetic studies has rapidly increased in recent years, there has been growing concern that the privacy of the participants in such studies can be invaded unless effective measures are adopted to protect confidentiality. It is crucial for the scientific community to establish a method to anonymize DNA samples so that the public will trust genetic researchers. Here, we present a reliable and practical method of making DNA samples used in the genetic research anonymous. It assures complete anonymity by coding samples and personal information twice. Since it does not require equipment, such as bar-code readers or a software package, its cost is nominal compared with the laboratory costs. All institutions engaged in genetic research may wish to take measures such as the one described here to ensure the privacy and confidentiality of the participants in their genetic studies. PMID- 12750963 TI - Association of positional and functional candidate genes FGF1, FBN2, and LOX on 5q31 with intracranial aneurysm. AB - We previously performed a genome-wide linkage study of intracranial aneurysm (IA) and found positive evidence of linkage at chromosomes 5q22-31, 7q11, and 14q22. In the present study, we focus on 5q31, where three candidate genes, fibroblast growth factor 1 (FGF1), fibrillin 2 (FBN2), and lysyl oxidase gene ( LOX) lie, and evaluate associations with IA. Genomic DNAs were obtained from 172 IA patients and 192 controls. Association analysis was performed with ten, five, and four single-nucleotide polymorphisms (SNPs) identified in FGF1, FBN2, and LOX, respectively. A difference in allelic frequency was observed for only the SNP at intron 4 in FGF1 (chi(2)=4.44, df=1, P=0.035). Although a haplotype association was observed with the combination of ten SNPs in FGF1 (chi(2)=16.04, df=1, P=0.00006), significant haplotype associations were not observed when haplotypes were constructed with the three, two, and four SNPs in FGF1 according to the linkage disequilibrium structure. No associations of FBN2 and LOX with IA were detected in the present study. PMID- 12750966 TI - Addition theorems for Slater-type orbitals and their application to multicenter multielectron integrals of central and noncentral interaction potentials. AB - By the use of complete orthonormal sets of psi(alpha)-ETOs (alpha=1, 0, m1, m2,...) introduced by the author, new addition theorems are derived for STOs and arbitrary central and noncentral interaction potentials (CIPs and NCIPs). The expansion coefficients in these addition theorems are expressed through the Gaunt and Gegenbauer coefficients. Using the addition theorems obtained for STOs and potentials, general formulae in terms of three-center overlap integrals are established for the multicenter t-electron integrals of CIPs and NCIPs that arise in the solution of the N-electron atomic and molecular problem (2hthN) when a Hylleraas approximation in Hartree-Fock-Roothaan theory is employed. With the help of expansion formulae for translation of STOs, the three-center overlap integrals are expressed through the two-center overlap integrals. The formulae obtained are valid for arbitrary quantum numbers, screening constants and location of orbitals. PMID- 12750965 TI - Structural analysis of the functional influence of the surface peptide Gtf-P1 on Streptococcus mutans glucosyltransferase C activity. AB - Glucosyltransferases (GtfB/C/D) in Streptococcus mutans are responsible for synthesizing water-insoluble and water-soluble glucans from sucrose and play very crucial roles in the formation of dental plaque. A monoclonal antibody against a 19-mer peptide fragment named Gtf-P1 was found in GtfC to reduce the enzyme activity to 50%. However, a similar experiment suggested almost unchanged activity in GtfD, despite of the very high sequence homology between the two enzymes. No further details are yet available to elucidate the biochemical mechanism responsible for such discrimination. For a better understanding of the catalytic behavior of these glucosyltransferases, structural and functional analyses were performed. First, the exact epitope was identified to specify the residue(s) required for monoclonal antibody recognition. The results suggest that the discrimination is determined solely by single residue substitution. Second, based on a combined sequence and secondary structure alignment against known crystal structure of segments from closely related proteins, a three-dimensional homology model for GtfC was built. Structural analysis for the region communicating between Gtf-P1 and the catalytic triad revealed the possibility for an "en bloc" movement of hydrophobic residues, which may transduce the functional influence on enzyme activity from the surface of molecule into the proximity of the active site. Figure Side chain interactions between Gtf-P1 and catalytic Asp 477 in GtfC. Calpha-tracing of GtfC with the two crucial peptides (Gtf-P1, orange; Gtf-P2, blue) and the catalytic triad residues ( red) highlighted to show their relative spatial organization. Side chains for the residues are also depicted according to their atom types. The structure is viewed with the barrel opening facing down PMID- 12750968 TI - Anion exchange in human serum transferrin N-lobe: a model study with variant His249Ala. AB - The removal of Fe(III) from human serum transferrin by chelators is thought to proceed through intermediate species in which the chelator becomes associated with the metal center of the protein. The visible spectral shifts associated with the formation of such intermediates in the wild-type (WT) protein are too small for reliable kinetic data to be obtained. Therefore, studies were undertaken with the recombinant N-terminal lobe variant H249A, a variant showing more pronounced spectral changes. The kinetics of the synergistic anion-exchange reaction between nitrilotriacetate (NTA) and carbonate in variant H249A was studied by stopped flow spectrophotometry as a model for this process in the WT protein. Anion exchange occurs by two pathways at pH 7.4 and 25 degrees C: an NTA-independent dissociative pathway to form a carbonate-free intermediate Fe-H249A (Eq. 1) that subsequently reacts with NTA (Eq. 2):and an NTA-dependent associative pathway (the major pathway) in which a quaternary Fe-H249A-(CO(3))(NTA) intermediate is formed (Eq. 3), which then decays to product (Eq. 4):The reverse reaction, where HCO(3)(-) exchanges for NTA, likewise follows these two pathways. The overall apparent equilibrium constant for formation of Fe-H249A-NTA from Fe-H249A-CO(3) is K'=442 at pH 7.4. The NTA complex is favored over the carbonate complex both kinetically and thermodynamically in the pH range 7.4-8.2. PMID- 12750967 TI - Resistance of marginal enamel to acid solubility is influenced by restorative systems: an in vitro scanning electron microscopic study. AB - The aim of this study was to evaluate the morphology of the enamel surface along margins of class V restorations following exposure to cariogenic solution. Restorations were placed in vitro in human third molars. The specimens were divided into groups according to resin composition: (1) Scotchbond 1 + Filtek Flow, (2) Scotchbond 1 + F2000, and (3) Prompt L-Pop + experimental flowable composite. Samples were stored in a demineralizing solution (lactic acid, pH 4.5, 0.1 M) at 37 degrees for 1-4 weeks or in deionized water (control group). The solution was changed every day. Replicas of the specimens were obtained in order to exclude drying artifacts. Scanning electron microscopy (SEM) of original and replica specimens identified a distinct enamel zone, defined as perimarginal enamel showing numerous fractures, porosities, voids, and pits. After the 4-week treatment, perimarginal prismatic enamel was greatly removed, while interprismatic enamel was still in place and only partially dissolved. Enamel not in relation with composite/compomer margins (0.5-1 mm away) showed minor alterations. Perimarginal enamel fractures probably due to composite/compomer shrinkage or the bur preparation may greatly contribute to this marginal enamel demineralization by increasing the number and size of porosities, that enhance the penetration and diffusion of cariogenic solution and create a sort of demineralized enamel subsurface. Only compomer restorations revealed a thin caries inhibition zone (1-2 micro m) probably related to fluoride release. Below this protected area, we observed the typical alterations of the other samples. These morphological alterations are probably related to secondary demineralization lesions and may affect the clinical life of restorations. PMID- 12750969 TI - Venous damage prevention by defibrotide in vinorelbine-treated patients. AB - GOALS: The aim of our study was to evaluate the incidence of venous toxicity induced by vinorelbine administration in patients who received a preventive therapy with defibrotide. PATIENTS AND METHODS: From July 1996 to July 2002 we treated 203 patients with vinorelbine, 51 with vinorelbine alone and 152 with vinorelbine in combination with other drugs via peripheral vein infusion. Of the 203 patients, 123 were male and 80 female with a median age of 67 years (range 18 to 82 years), and 118 were chemotherapy-naive. Defibrotide was delivered i.v. at a dose of 400 mg in 250 ml normal saline. After infusion of 125 ml over about 15 min, vinorelbine mixed with 10 ml normal saline was delivered as quick brief repeated pulses over 5 min through the plastic tube, followed by infusion of the remaining defibrotide. The specific Rittenberg scale was used to assess venous irritation episodes. RESULTS: Among a total of 1336 vinorelbine infusions, with a median of five infusions per patient, the incidence of venous irritation episodes graded according to Rittenberg scale was 1.1% (15), of which 0.6% (8) were grade 2 and 0.5% (7) grade 1. Globally, 15 patients (7.3%) developed venous toxicity after a median of 3 infusions (range 1-14), but no patient had more than one event. CONCLUSION: Our findings support the use of defibrotide as an effective, safe and low-cost means for preventing vinorelbine-related venous damage. PMID- 12750970 TI - Climatic factors governing plant phenological phases along a Norwegian fjord. AB - In the present project, the time of leaf budding and flowering, and partly also of fruit ripening, was studied over 3 years in different cultivated and native plants on a gradient along a western Norwegian fjord about 300 km long, from oceanic to relatively continental regions. In the plants investigated, flowering of the red currant was most strongly favoured by oceanic conditions in the outermost part of the fjord. On the other hand, flowering of the apple was earliest in the middle district, as were flowering of the common lilac and raspberry, while differences were small between the districts for flowering of the plum and pear. In the inner district, leaf budding of the apple was about 1.5 weeks earlier than flowering of the red currant, while these two phenophases, on average, occurred on the same day in the oceanic district. The time from 1 April to flowering was generally lengthened by increased precipitation in the pear, apple, lilac and raspberry, but not in the red currant and plum. By contrast, the period from leaf budding to flowering was significantly shortened in the plum by high precipitation. The present studies also indicated that leaf budding of the birch was favoured by the high minimum temperature and the relatively high precipitation normally found in the oceanic district. Partial correlations showed that increased precipitation delayed the flowering of both rowan and bird cherry trees; there was also a week effect on bud break in the same two species. The clear conclusion of the present study, therefore, is that various plant species react differently to various climatic factors ("phenological interception"), even in different phenophases within the same species. This means that the various species are best fitted to certain climatic regions and should preferably be planted there if other growth factors are satisfactory. PMID- 12750971 TI - Heat stress and mortality in Lisbon Part II. An assessment of the potential impacts of climate change. AB - Global environmental change, in particular climate change, will have adverse effects on public health. The increased frequency/intensity of heat waves is expected to increase heat-related mortality and illness. To quantify the climatic risks of heat-related mortality in Lisbon an empirical-statistical model was developed in Part I, based on the climate-mortality relationship of the summer months of 1980-1998. In Part II, scenarios of climate and population change are applied to the model to assess the potential impacts on public health in the 2020s and 2050s, in terms of crude heat-related mortality rates. Two regional climate models (RCMs) were used and different assumptions about seasonality, acclimatisation and the estimation of excess deaths were made in order to represent uncertainty explicitly. An exploratory Bayesian analysis was used to investigate the sensitivity of the result to input assumptions. Annual heat related death rates are estimated to increase from between 5.4 and 6 (per 100,000) for 1980-1998 to between 5.8 and 15.1 for the 2020s. By the 2050s, the potential increase ranges from 7.3 to 35.6. The burden of deaths is decreased if acclimatisation is factored in. Through a Bayesian analysis it is shown that, for the tested variables, future heat-related mortality is most sensitive to the choice of RCM and least to the method of calculating the excess deaths. PMID- 12750973 TI - A sporadic case of cystinuria, respiratory chain and growth hormone deficiencies. PMID- 12750972 TI - Effects of head cooling on human sleep stages and body temperature. AB - The purpose of this study was to confirm the effect of head cooling on human sleep stages and body temperature. Nine healthy male volunteers with a mean age of 25 +/- 3.77 years served as subjects. The experiments were carried out under three different sets of conditions: 26 degrees C, relative humidity (RH) 50% (26/50); 32 degrees C, RH 80% (32/80); and 32 degrees C RH 80% with the use of a cooling pillow (32/80 HC). The subjects slept from 2300 hours to 0700 hours with a cotton blanket, wearing short-sleeved pyjamas and shorts on a bed, which was covered with a sheet. Electroencephalograms, electro-ouclogram, and mental electromyelograms were recorded through the night. Rectal temperature (Tre) and skin temperature (Tsk) were measured continuously. Whole-body sweat and the tympanic temperature (Tty) were measured before and after sleep. Wakefulness significantly increased at 32/80 than at 26/50; however, no significant difference was observed between 32/80 HC and 26/50. Tre and mean Tsk were higher both at 32/80 and 32/80 HC than at 26/50. The whole-body sweat loss was significantly greater and Tty in the morning was higher at 32/80 than 32/80 HC and 26/50. These results suggest that head cooling during sleep may help to decrease the whole-body sweat rate during sleep under humid heat conditions. PMID- 12750974 TI - Another factor influencing the urinary calcium excretion. PMID- 12750975 TI - Cyclophosphamide in steroid-sensitive nephrotic syndrome: outcome and outlook. AB - Steroid-sensitive nephrotic syndrome often follows a relapsing course with a substantial number of patients requiring cytotoxic therapy with cyclophosphamide (CP). However, the long-term success of CP treatment is difficult to predict. We retrospectively evaluated 106 patients after CP to identify parameters associated with sustained remission. The overall rate of cumulative sustained remission was 24% after 10 years. No gender difference was found. Several factors were significantly correlated with the rate of sustained remission: age at CP therapy (remission 34% versus 9% in children older or younger than 5.5 years, P<0.01), frequently relapsing versus steroid-dependent status (54% versus 17%, P<0.05), leukopenia under CP treatment (44% in children with leukopenia versus 19% in children without leukopenia, P<0.05), and a cumulative dosage per body surface area (BSA) of more or less than 5,040 mg/m(2) (45% versus 11%, P<0.01). In contrast, the cumulative dosage per kilogram body weight had no significant influence on long-term remission (23% in children with >168 mg/kg versus 26% in children with <168 mg/kg, P>0.05). The current concept of CP treatment of steroid sensitive nephrotic syndrome is less effective in preschool children. CP therapy should be re-evaluated on a BSA-adjusted regimen. PMID- 12750976 TI - Pamidronate in a girl with chronic renal insufficiency dependent on parenteral nutrition. AB - A 10-year-old 40-kg African-American female with megacystis microcolon hypoperistalsis syndrome maintained on total parenteral nutrition (TPN), with a history of metabolic bone disease and renal insufficiency, was admitted with a Candida parapsilosis central venous line infection. During her 280-day hospital stay, she had multiple episodes of bacteremia and recurrent candidemia. Furthermore, she developed pathological fractures and hip displacement with osteomyelitis due to Enterobacter. Hypercalcemia and a history of nephrocalcinosis had prevented appropriate dosing of calcium prior to and during the first months of her hospital stay. Pamidronate and chlorothiazide were added to her regimen. The urinary calcium to creatinine ratio and ionized calcium decreased. The pamidronate dose was increased to 60 mg once a week and was well tolerated. Daily calcium was added to her TPN solution and was increased to 10 mEq/day by the time of discharge. We conclude that relatively large doses of pamidronate may be required in certain cases of refractory hypercalcemia and are well tolerated in children. PMID- 12750977 TI - Association of hypocalcemia with a change in gentamicin administration in neonates. AB - Aminoglycosides are administered frequently to neonates with suspected sepsis. We report the association of hypocalcemia in term and near-term neonates receiving gentamicin therapy for >/=4 days after a change in dosing from every 12 h to every 24 h. The possible association with a higher gentamicin dose and longer dosing interval is described. PMID- 12750979 TI - Nephrocalcinosis and medullary cysts in 3-methylglutaconic aciduria. AB - 3-methylglutaconic aciduria is frequently found during urine organic acid analysis and is widely regarded as a marker of a mitochondrial disorder, the clinical features of which are very heterogeneous. We describe two siblings with 3-methylglutaconic aciduria in whom renal ultrasonography showed echogenic medullae consistent with nephrocalcinosis. One patient also developed medullary cysts. In both children renal function was normal and neither had any plasma or urinary evidence of tubulopathy. The presence of nephrocalcinosis and medullary cysts in patients with 3-methylglutaconic aciduria adds to the heterogeneous clinical presentation of this group of disorders. PMID- 12750978 TI - Epstein-Barr virus-associated acute renal failure: diagnosis, treatment, and follow-up. AB - We retrospectively reviewed our experience of Epstein-Barr virus (EBV)-associated acute renal failure. Of 165 previously healthy children hospitalized with serologically proven primary EBV infection, 8 had acute renal failure, of whom 5 (group A) did not have virus-associated hemophagocytic syndrome (VAHS), while 3 (group B) did have VAHS. All had complications in four or more organ systems. Two patients in group A had renal biopsies showing acute tubulointerstitial nephritis, and the clinical and laboratory findings in the other 3 group A patients were consistent with acute tubulointerstitial nephritis. Acyclovir was used in 1 patient, but she died of hepatic failure and pulmonary hemorrhage. The other 4 spontaneously recovered renal function after supportive care, including hemodialysis in 1 patient. Our experience does not support the routine use of corticosteroids or antiviral agents in these patients. Children in group B had a relatively normal urinalysis. Renal biopsies were not performed, but their presentations were compatible with acute tubular necrosis. We conclude that EBV should be considered as a possible etiological agent in all children presenting with acute renal failure of unknown cause. The diagnosis depends on a high index of suspicion and careful serological evaluation in atypical cases. PMID- 12750980 TI - Chronic renal failure in children: a report from Port Harcourt, Nigeria (1985 2000). AB - A 15-year review of children who presented with chronic renal failure (CRF) to the University of Port Harcourt Teaching Hospital, Rivers State of Nigeria, was carried out. Forty-five children (28 boys, 17 girls) with CRF, defined as a glomerular filtration rate below 30 ml/min per 1.73 m(2) body surface area or a rise in serum creatinine above 120 micro mol/l for at least 6 months, often accompanied by other biochemical abnormalities, were identified. The median annual incidence of CRF was 3.0 per million children. The prevalence of CRF increased from 12.5 in the 1985-1990 periods to 15 per million children after 1995. Acquired disorder was the major cause of CRF. Glomerulopathies were the cause in 53.3% of patients, mainly chronic glomerulonephritis (56.5%) and nephrotic syndrome (30.4%). Hepatitis B surface antigen was positive in 2 patients. Congenital disorders accounted for 28.9% of all cases of CRF, which is lower than data from other countries. Posterior urethral valve was the only congenital disorder causing CRF in the study. No child with hereditary renal disorder as a cause of CRF was identified. Children with congenital disorders were diagnosed at an earlier age. The mortality rate was high (46.7%), as most patients were managed conservatively, since there were no permanent facilities for chronic dialysis or renal transplantation in Nigeria. The study shows that CRF is common in Nigerian children, and there is an urgent need for the establishment of facilities for renal replacement therapy. PMID- 12750981 TI - The effect of dopamine on glomerular filtration rate in normotensive, oliguric premature neonates. AB - We examined the effect of dopamine on glomerular filtration rate (GFR) at infusion rates of 0.5, 2.5, and 7.5 micro g/kg per min in 15 premature neonates. Study infants (mean gestational age 34+/-2 weeks, mean birth weight 2.43+/-0.6 kg) had respiratory distress, were normotensive, and had a low urine output (0.9+/-0.1 ml/kg per hour). GFR was determined by the plasma clearance of inulin after a single bolus injection (200 mg/kg). Four hours after inulin administration, dopamine infusion was begun and continued over 6 h. GFR was estimated before and after beginning the dopamine infusions from the slope of the log of plasma inulin concentration versus time. Gestational age, weight, and baseline GFR were similar in all three groups. With a dopamine infusion rate of 0.5 micro g/kg per min there were no changes in GFR, urine output, heart rate, or blood pressure. At an infusion rate of 7.5 micro g/kg per min there was no change in GFR, although urine output, heart rate, and blood pressure all increased. At 2.5 micro g/kg per min there were significant increases in GFR and urine output, with no changes in blood pressure or heart rate. In oliguric, non-hypotensive neonates, GFR increased significantly at 2.5 micro g/kg per min of dopamine. This probably reflects the effects of afferent vasodilatation and may be important clinically when enhancement of GFR is the major treatment objective. PMID- 12750982 TI - Final height in children with chronic renal failure who have not received growth hormone. AB - Final height (FH), growth velocity after 16 and 18 years of age, and factors predictive for FH were assessed in 60 patients (21-36 years old), whose chronic renal failure (CRF) started before the age of 16 years (28 girls and 32 boys). At 16 years of age, 22 had conservative treatment (CT, group A) and 38 end-stage renal failure [ESRF, group B, which includes 19 receiving hemodialysis (HD) and 19 with a functional renal transplant (RTx)]. None received recombinant human growth hormone (rhGH) treatment. FH was lower than in a normal population: 161.6+/-8 cm for males [-2.06+/-1.3 standard deviation score (SDS)] and 154.3+/ 8.1 cm for females (-1.4+/-1.4 SDS). FH in group A (-1.15+/-1.4 SDS) was significantly higher than in group B (-2.1+/-1.3 SDS); 45% of all patients (56% of males and 23% of females) had a final height below -2 SDS (41% in group A and 47% in group B). FH was reached at 20.2+/-1.8 years in males and 18.8+/-2 years in females. A continuation of growth after 18 years of age was observed in 23 males (71.8%): +5.2 cm (+0.87 SDS) and in 14 females (50%): +1.75 cm (+0.3 SDS). However, this partial recovery concerned mainly patients with an important growth deficiency. A higher height at enrolment or at ESRF was significantly associated with a higher FH, whereas a longer period of ESRF had a significantly negative effect. In conclusion, all efforts should be made to diagnose CRF as early as possible and to try to improve growth before ESRF and RTx. Early institution of rhGH therapy should improve FH and improve the chance of achieving near-normal adult height in most patients. PMID- 12750983 TI - Lip hypertrophy secondary to cyclosporin treatment. AB - Gingival hypertrophy is a well-documented side effect of cyclosporin therapy, but severe lip enlargement is less frequently recognised. This can lead to poor body image, low self-esteem and non-compliance, especially in the older childhood and adolescent age groups. We describe two paediatric renal transplant recipients who had marked lip hypertrophy as a consequence of cyclosporin (Neoral) treatment. On changing the immunosuppression to tacrolimus (Prograf), this resolved. We recommend that a change in immunosuppressant therapy be considered in children with significant changes to facial appearance. PMID- 12750984 TI - Growth in steroid-responsive nephrotic syndrome: a study of 85 pediatric patients. AB - The statural growth of 85 patients with steroid-responsive idiopathic nephrotic syndrome, attending the Pediatric Nephrology Unit, Children's Institute, Hospital das Clinicas School of Medicine, University of Sao Paulo, with a minimum follow up of 3 years, was evaluated. Analysis of the patient population as a group did not show any significant alterations in the height Z score and the mean height percentile between the first (-0.59 and 33.9, respectively) and last consultation (-0.57 and 34.8, respectively). Analysis of each individual patient allowed the definition of two subgroups. Subgroup A, which achieved growth improvement, was composed of 47 children-initial Z score and mean initial height percentile of 0.91 and 24.0, respectively; final Z score and mean height percentile of -0.30 and 40.7, respectively ( P=0.00). Subgroup B, which showed growth retardation, was composed of 38 children-initial Z score and mean initial height percentile of -0.19 and 46.2, respectively; final Z score and mean height percentile of -0,9 and 27.5, respectively ( P=0.00). The following factors were significantly different when both subgroups were compared: (1) total duration of prednisone therapy and total prednisone dose were greater in subgroup B; (2) the final chronological age of patients using prednisone was higher in subgroup B; (3) the pubertal growth spurt in subgroup B showed attenuation and retardation. PMID- 12750986 TI - Increased c-Met tyrosine kinase expression in segmental renal dysplasia. AB - Renal dysplasia (RD) is a disorganized development of renal parenchyma that results in a deficit of functional renal tissue. It has been suggested in the animal model that increased expression of HGF receptor, c-Met tyrosine kinase in the epithelial cells during kidney development may induce a growth of dysplastic epithelia and result in RD. The aim of this study was to investigate the immunoreactivity of c-Met tyrosine kinase in the dysplastic kidney in order to further understand the pathogenesis of RD. Specimens of dysplastic upper pole kidney were obtained from 19 patients during upper pole partial nephrectomy for non-functioning upper moiety of duplex kidney. In the dysplastic kidney, there was strong c-Met immunoreactivity in the epithelium of primitive tubules. In contrast, c-Met immunoreactivity was barely detectable in the normal kidney. Markedly increased expression of HGF receptor, c-Met tyrosine kinase in renal dysplasia suggests that HGF may be involved in the development of renal dysplasia. PMID- 12750987 TI - Acute leukemia: an association with atypical hemolytic uremic syndrome. AB - A 5-year-old boy was admitted for anemia, thrombocytopenia, and azotemia. Bone marrow examination demonstrated only erythroid hyperplasia, and he was diagnosed with an atypical form of hemolytic uremic syndrome (HUS). He was treated with daily prednisolone, and hematological profiles and renal function normalized 1 month later. However, heavy proteinuria persisted, and renal biopsy findings were consistent with HUS. Four months later, pancytopenia developed with recurrence of azotemia, and a second bone marrow examination revealed acute lymphoblastic leukemia (ALL). The remission of proteinuria with no recurrences of HUS after antileukemic treatment suggests that the HUS was associated with the ALL. PMID- 12750985 TI - The management of anemia in pediatric peritoneal dialysis patients. Guidelines by an ad hoc European committee. AB - Anemia is common in chronic renal failure. Guidelines for the diagnosis and treatment of anemia in adult patients are available. With respect to the diagnosis and treatment in children on peritoneal dialysis, the European Pediatric Peritoneal Dialysis Working Group (EPPWG) has produced guidelines. After a thorough diagnostic work-up, treatment should aim for a target hemoglobin concentration of at least 11 g/l. This can be accomplished by the administration of erythropoietin and iron preparations. Although there is sufficient evidence to advocate the intraperitoneal administration of erythropoietin, most pediatric nephrologists still apply erythropoietin by the subcutaneous route. Iron should preferably be prescribed as an oral preparation. Sufficient attention has to be paid to the nutritional intake in these children. There is no place for carnitine supplementation in the treatment of anemia in pediatric peritoneal dialysis patients. PMID- 12750988 TI - A de novo R589C mutation of anion exchanger 1 causing distal renal tubular acidosis. AB - Anion exchanger 1 (AE1 or SLC4A1) mutations have been reported to cause distal renal tubular acidosis (dRTA), a disease characterized by impaired acid excretion in the distal nephron. We have recently demonstrated homozygous AE1 G701D mutation as a common molecular defect of autosomal recessive (AR) dRTA in a group of Thai pediatric patients. In the present work, we discovered a de novo heterozygous AE1 R589C mutation, previously documented in inherited autosomal dominant (AD) dRTA. Arginine at this position is conserved in all vertebrate AE proteins indicating its functional importance. Three different mutations at this position (R589C, R589H, and R589S) were all found in AD dRTA and a de novo R589H mutation has previously been recorded. Our report is the second de novo mutation but with a different substituted amino acid. A high prevalence of AE1 R589 mutations and the presence of at least two de novo mutations at this position lead us to propose that codon 589 (CGC) is a "mutational hotspot" of AE1. The mechanism of recurrent mutations probably involves methylation and deamination altering cytosine (C) to thymine (T) in the CpG dinucleotides. PMID- 12750990 TI - Seasonal variation in top-down and bottom-up processes in a grassland arthropod community. AB - Both top-down and bottom-up processes are common in terrestrial ecosystems, but how these opposing forces interact and vary over time is poorly understood. We tested the variation of these processes over seasonal time in a natural temperate zone grassland, a field site characterized by strong seasonal changes in abiotic and biotic conditions. Separate factorial experiments manipulating nutrients and cursorial spiders were performed in the wet and dry seasons. We also performed a water-addition experiment during the summer (dry season) to determine the degree of water limitation during this time. In the spring, nutrient addition increased plant growth and carnivore abundance, indicating a bottom-up control process. Among herbivores, sap-feeders were significantly enhanced while grazers significantly declined resulting in no net change in herbivore abundance. In the summer, water limitation was predominant increasing plants and all herbivores while nutrient (N) effects were non-significant. Top-down processes were present only in the spring season and only impacted the guild of grazing herbivores. These results show that bottom-up limitation is present throughout the season in this grassland, although the specific limiting resource changes as the season progresses. Bottom-up processes affected all trophic levels and many different guilds, while top-down effects were limited to a select group of herbivores and did not extend to the plant trophic level. Our results show that the relative strengths of top-down and bottom-up processes can shift over relatively short periods of time in habitats with a strong seasonal component. PMID- 12750989 TI - Temporal variation in delta(13)C of ecosystem respiration in the Pacific Northwest: links to moisture stress. AB - We measured seasonal and interannual variations in delta(13)C values within the carbon reservoirs (leaves and soil) and CO(2) fluxes (soil and ecosystem respired CO(2)) of an old growth coniferous forest in the Pacific Northwest USA with relation to local meteorological conditions. There were significant intra-annual and interannual differences in the carbon isotope ratios of CO(2) respired at both the ecosystem (delta(13)C(R)) and the soil levels (delta(13)C(R-soil)), but only limited variations in the carbon isotope ratios of carbon stocks. The delta(13)C(R) values varied by as much as 4.4 per thousand over a growing season, while delta(13)C(R-soil )values changed as much as 6.2 per thousand. The delta(13)C of soil organic carbon (delta(13)C(SOC)) and needle organic carbon (delta(13)C(P)) exhibited little or no significant changes over the course of this study. Carbon isotope discrimination within leaves (Delta(p)) showed systematic decreases with increased canopy height, but remained fairly constant throughout the year (Delta(p)=17.9 per thousand -19.2 per thousand at the top of the canopy, Delta(p)=19.6 per thousand -20.9 per thousand at mid-canopy, Delta(p)=23.3 per thousand -25.1 per thousand at the canopy base). The temporal variation in the delta(13)C of soil and ecosystem respired CO(2) was correlated ( r=0.93, P<0.001) with soil moisture levels, with dry summer months having the most (13)C-enriched values. The dynamic seasonal changes in delta(13)C of respired CO(2) are hypothesized to be the result of fast cycling of recently fixed carbon back to the atmosphere. One scaling consequence of the seasonal and interannual variations in delta(13)C(R) is that inversion-based carbon-cycle models dependent on observed atmospheric CO(2) concentration and isotope values may be improved by incorporating dynamic delta(13)C(R) values to interpret regional carbon sink strength. PMID- 12750991 TI - Contribution of relative growth rate to root foraging by annual and perennial grasses from California oak woodlands. AB - Plants forage for nutrients by increasing their root length density (RLD) in nutrient-rich soil microsites through root morphological changes resulting in increased root biomass density (RBD), specific root length (SRL), or branching frequency (BF). It is commonly accepted that fast-growing species will forage more than slow-growing species. However, foraging responses may be due solely to differences in relative growth rates (RGR). There is little evidence, after the effects of RGR are removed, that the fast versus slow foraging theory is correct. In a pot study, we evaluated foraging of four grass species that differed in RGR: one fast-growing annual species, Bromus diandrus, two intermediate-growing species, annual Bromus hordeaceus and perennial Elymus glaucus, and one slow growing perennial species, Nassella pulchra. We harvested plants either at a common time (plants varied in size) or at a common leaf number (plants similar size, surrogate for common biomass). By evaluating species at a common time, RGR influenced foraging. Conversely, by evaluating species at a common leaf number, foraging could be evaluated independent of RGR. When RGR was allowed to contribute to foraging (common time harvest), foraging and RGR were positively correlated. B. diandrus (fast RGR) foraged to a greater extent than did E. glaucus (intermediate RGR) and N. pulchra (slow RGR). E. glaucus (intermediate RGR) foraged to a greater extent than N. pulchra (slow RGR). Root growth within nutrient-rich microsites was due to significant increases in RBD, not to modifications of SRL or BF. However, when RGR was not allowed to influence foraging (common leaf number harvest), none of the four species significantly enhanced RLD in nutrient-rich compared to control microsites. This suggests that RGR strongly influenced the ability of these grass species to forage and also supports the need to evaluate plastic root traits independent of RGR. PMID- 12750992 TI - Nitrogen fixation and nitrogen limitation of primary production along a natural marsh chronosequence. AB - Nitrogen (N) limitation of primary production is common in temperate salt marshes, even though conservative N recycling can fulfill a large proportion of plant N demand. In nutrient poor young marshes, N limitation may be more severe and new N sources, such as N fixation, more important for plant growth. We measured N fixation and the response of salt marsh primary producers (Spartina alterniflora and benthic microalgae) to N fertilization in one mature (>150 year) and two young (7 and 15 year) naturally developing marshes at the Virginia Coast Reserve LTER site. S. alterniflora aboveground biomass in the mature marsh (1,700+/-273 g m(-2)) was 1.8 and 2.8 times higher than in the 15 year and 7 year old marshes, respectively. Fertilization significantly increased S. alterniflora biomass in the two young marshes (160-175%) and areal aboveground tissue N in the youngest marsh (260%). Microalgal chlorophyll a (Chl a) in the mature marsh was nearly 2-fold lower than in the 7-year-old marsh, and there was no evidence that this was due to light limitation. However, Chl a in fertilized plots was 30% higher than control plots at the youngest site. Daily N fixation decreased with increasing marsh age in summer, when rates were highest at all sites. Autotrophic N fixation (difference between rates in the light and dark) was most important in the summer, but we saw no indication of a shift in dominance between autotrophic and heterotrophic N fixers during marsh development. Estimated annual N fixation was 2- to 3-fold higher in the young marsh (18.3+/-1.5 g N m(-2) year(-1)), than in the intermediate-aged (9.0+/-0.7) or mature marsh (6.1+/-0.5). In the young marshes, N fixation was sufficient to provide a substantial proportion of aboveground S. alterniflora N demand. Our results suggest that both benthic microalgae and S. alterniflora in young salt marshes are N limited, and that this limitation decreases as the marsh matures. The high rates of N fixation by autotrophic and heterotrophic bacteria in the sediment could provide an important source of N for primary producers during marsh development. PMID- 12750993 TI - Effects of pregnancy on locomotor performance: an experimental study on lizards. AB - Pregnancy is associated with reduced locomotor performance in many types of animals, but we do not know to what degree this correlation is caused by simple physical burdening, versus physiological changes associated with pregnancy, or to confounding variables (such as season or female body size) that simultaneously influence both reproductive investment and locomotor speeds. To identify causal effects of burden on locomotion, we need to experimentally manipulate the size of the load being carried. Injection of sterile fluid into the peritoneal cavities of 84 garden skinks (Lampropholis guichenoti) showed that speeds decreased with increasing burdens. Lizards with a burden equivalent to 25% of their body mass ran about 15% slower, mirroring the situation seen in gravid lizards of this population. Thus, simple physical burdening appears to be the primary causal component of the locomotor cost of reproduction within these animals. A lizard's sex, body size and shape had little effect on its running ability either before or after treatment, but faster lizards showed a greater performance decrement after burdening than did their slower conspecifics. PMID- 12750995 TI - Lethal effect of ammonia on metacercariae of Clonorchis sinensis. AB - The effect of endogenous ammonia produced in dead fish or ammonia in ammonium bicarbonate solutions was evaluated on the viability of Clonorchis sinensis metacercariae. Viability was determined by worm recovery in rats infected with metacercariae exposed to ammonia in decaying fish or in vitro solutions. The recovery rates were 58%, 48%, 44% and 2% in fish that had been allowed to decay for 5, 10, 20 and 30 days at 4 degrees C, respectively, and this rate ( Y) was inversely correlated with the ammonia concentration ( X) in fish muscle extract ( Y=80.62-0.0667 X, r(2)=0.905, P=0.049). C. sinensis metacercariae barely survived in ammonia solutions with concentrations exceeding 1 g N/l. Our results indicate that the ammonia produced in dead fish is lethal to the viability of C. sinensis metacercariae. PMID- 12750994 TI - Angiostrongylus costaricensis infection in C57BL/6 mice: MHC-II deficiency results in increased larval elimination but unaltered mortality. AB - During experimental Angiostrongylus costaricensis infections in several inbred mouse strains, genetic factors as well as different cytokine secretion patterns have recently been shown to play a role in the outcome of infection in terms of morbidity and mortality, e.g. BALB/c mice show a high and C57BL/6 mice a low mortality during the acute phase of infection. In this study, C57BL/6 MHC-II knockout mice infected with A. costaricensis did not show increased mortality during the acute phase of infection when compared with wild-type mice. Furthermore, MHC-II knockout mice showed a strongly diminished parasite-specific humoral and cellular immune response, which can be explained by the nearly complete lack of CD4+ T cells in the periphery. This defect in MHC-II genes, the lack of CD4+ T cells, and the resulting cellular and humoral unresponsiveness resulted in a three times higher output of first-stage larvae in feces compared with wild-type animals. The results indicate that during experimental A. costaricensis infection a parasite-specific immune response, directed via MHC-II molecules and CD4+ T cells, is not essential for the survival of C57BL/6 mice during the acute phase of infection, whereas the elimination of first-stage larvae seems to be regulated by a MHC-II- and CD4+ T-cell-dependent mechanism. PMID- 12750996 TI - Classification of Dukes' B and C colorectal cancers using expression arrays. AB - PURPOSE: Colorectal cancer is one of the most common malignancies. Substaging of the cancer is of importance not only to prognosis but also to treatment. Classification of substages based on DNA microarray technology is currently the most promising approach. We therefore investigated if gene expression microarrays could be used to classify colorectal tumors. METHODS: We used the Affymetrix oligonucleotide arrays to analyze the expression of more than 5,000 genes in samples from the sigmoid and upper rectum of the left colon. Five samples were from normal mucosa and five samples from each of the Dukes' stages A, B, C, and D. Expression data were filtered based on either covariance or a selection of the most significantly varying genes between tumor stages. RESULTS: A nearest neighbor classifier was used to classify normal, and Dukes' B and C samples with less than 20% error, whereas Dukes' A and D could not be classified correctly. A number of interesting gene clusters showed a discriminating difference between Dukes' B and C samples. These included mitochondrial genes, stromal remodeling genes, and genes related to cell adhesion. CONCLUSION: Molecular classification based on gene expression of one of the most common malignancies, colorectal cancer, now seems to be within reach. The data indicates that it is possible at least to classify Dukes' B and C colorectal tumors with microarrays. PMID- 12750997 TI - Indium-111-Octreotide scintigraphy in differentiated thyroid carcinoma metastases that do not respond to treatment with high-dose I-131. AB - AIM: Differentiated thyroid cancer is characterized by a very good prognosis in the majority of the patients. The therapy of choice is surgery followed by ablation with Iodine-131 (I-131). However, some patients have metastases that have lost the capability of concentrating I-131, even when it is given in therapeutic doses. In the present study, we describe the value of Indium-111 Octreotide (Octreoscan) in differentiated thyroid cancer patients with increased Tg levels who failed to demonstrate a response to treatment with high-dose I-131. METHOD: Fifteen consecutive patients with progressive differentiated thyroid cancer (ten female, five male) (mean age: 59 years, range 13-81 years; eight papillary, six follicular, and one Hurthle cell carcinoma) were studied. Progression was based on increasing Tg levels and was confirmed by radiological evaluation. Whole body scintigraphy (WBS) was performed after the administration of 200 MBq of Indium-111-Octreotide. The images were assessed by two experienced observers and compared with post-treatment I-131 WBS. RESULTS: In seven out of 15 patients distant metastases were already present at initial stage, whereas in ten patients the primary tumor stage was T3 or T4 indicating that the majority of the patients had advanced disease. In two out of five patients with a positive I-131 WBS, Indium-111-Octreotide was false negative. In nine out of ten patients with a negative I-131 WBS, Indium-111-Octreotide demonstrated multiple metastases. In those patients with a positive SSR-scintigraphy, metastases were found in the lungs ( n=14), bone ( n=7), mediastinum ( n=3), liver ( n=2), brains ( n=1), and cutis ( n=1). Overall, three out of 15 patients had a negative Indium-111 Octreotide result revealing a sensitivity of 80%. CONCLUSION: Our findings demonstrate the diagnostic value of Indium-111-Octreotide in differentiated thyroid cancer that fails to respond to I-131 treatment. It opens the possibility for additional treatment with high doses of Indium-111-Octreotide or its analogs in a majority of the patients. PMID- 12750998 TI - The abnormalities of chromosome 8 in two hepatocellular carcinoma cell clones with the same genetic background and different metastatic potential. AB - PURPOSE: Two hepatocellular carcinoma (HCC) cell clones named MHCC97-H and MHCC97 L with different metastatic potential have recently been established from the same parent cell line MHCC97 in our institute. The cytogenetic alterations of these two clones were investigated in this study to explore the possible clues to the mechanism involved in HCC metastasis. METHODS: Their chromosomal aberrations were analyzed with comparative genomic hybridization (CGH), chromosome-specific painting, and two-color fluorescence in situ hybridization (FISH). RESULTS: The aberrations were found in a total of 17 chromosomes, and six kinds of the aberrations including gains of 1q, 7q, 8q, 20, and the losses of 8p23, 21q were found both in the two cell clones and their parent cell line MHCC97. Using modified CGH, with the DNA of MHCC97-L as control to test the MHCC97-H clone, the loss of 8p23 and the gain of 1q31-32, 8q21.3-22, 13q22, 17q22 were highlighted, and the most significant finding was on chromosome 8. Dual color FISH combining a pericentromeric probe and a BAC probe mapping at 8q23.1 was then performed to verify this result, and the signal ratios of the BAC to centromere were 1.43 in MHCC97-H and 1.45 in MHCC97-L, confirming the over-representations at 8q in both cells. Another interesting finding in the dual-color metaphase FISH was the intrachromosomal translocation of 8q to 8p (looked like an isochromosome 8) and non-reciprocal translocation of part of 8q to 4q, which was further clarified and proved by the FISH with whole chromosome 8 painting probe. CONCLUSIONS: The high copies amplification on 8q, the formation of isochromosome 8, non-reciprocal translocation of partial 8q to 4q, and loss of 8p occurred at the same time and are the characteristic chromosomal aberrations of the two cell clones. The chromosome 8p, especially 8p23, might harbor some novel genes related to the HCC metastasis. PMID- 12750999 TI - Urinary lithogenic and inhibitory factors in preterm neonates receiving either total parenteral nutrition or milk formula. AB - The aim of this study was to evaluate prospectively the influence of nutrition on certain factors which may inhibit or promote nephrocalcinosis in two groups of preterm infants, receiving total parenteral nutrition (TPN) and special preterm milk formula respectively, but not furosemide. A total of 37 preterm infants, 15 on TPN and 22 fed a special preterm formula were studied at the end of the 1st, 2nd and 3rd weeks of life, at which time serum and 8 h urine specimens were collected. High ratios of urinary calcium to urinary creatinine (UCa/cr), urinary oxalate to urinary creatinine (Uox/cr) and urinary calcium to urinary citrate (UCa/cit) indicates an increased risk for nephrocalcinosis while high urinary citrate to urinary creatinine (Ucit/cr) ratio indicates protection. Uox/cr increased significantly (P<0.05) in those infants fed preterm formula, from the end of 2nd week of life and was two-fold higher than in the TPN group of preterm infants (P<0.01). Ucit/cr was higher throughout the study period in the formula fed than in the TPN preterm infants. UCa/cit was five-fold higher (P<0.01) in the TPN group, by the end of the 3rd week. Urinary calcium and magnesium was similar in both groups during the study period. Two of the infants studied (5.4%), one from each group, developed nephrocalcinosis. CONCLUSION: In preterm neonates on total parenteral nutrition, urinary oxalate -to-creatinine ratio (a potent lithogenic factor) was lower and urinary citrate -to-creatinine ratio (a lithoprotective factor) also lower than in formula fed neonates. The type of feeding (total parenteral nutrition or special preterm milk formula) seems to affect urinary oxalate and citrate but not calcium and magnesium in non furosemide treated preterm infants during the first 3 weeks of life. PMID- 12751001 TI - Risk factors for neonatal tetanus in Ibadan, Nigeria. PMID- 12751000 TI - Familial mediterranean fever: revisiting an ancient disease. AB - Familial Mediterranean fever (FMF) is an auto-inflammatory disease characterised by periodic attacks of fever and serositis. Recent genetic and epidemiological research have highlighted the importance of this disease. FMF is the most frequent periodic fever syndrome and is transmitted in an autosomal recessive fashion. The disease is caused by mutations in the gene on the short arm of chromosome 16, coding for the protein "pyrin". Pyrin is mainly expressed in neutrophils and monocytes and is among the proteins involved in the interleukin-1 inflammatory pathway. The recurrent attacks of fever are accompanied by severe abdominal pain, arthritis and/or chest pain along with a marked increase in acute phase reactants. Among these, serum amyloid A protein is especially important since it is the precursor of the amyloid A fibrils deposited in secondary renal amyloidosis. Renal amyloidosis has a grave prognosis. Differential diagnosis from other periodic fever syndromes is especially important in western European countries. Among these hyper IgD syndrome is common in Netherlands and the tumour necrosis factor receptor-associated periodic syndrome is especially common among Scottish and Irish families. Mutation analysis of the gene may be helpful in diagnosing FMF; however, if this is not possible, a trial of colchicine is a helpful diagnostic tool. The indications for life-long colchicine treatment should be discussed with the family. CONCLUSION: Familial mediterranean fever and other auto-inflammatory syndromes should be suspected in children with recurrent febrile attacks. Early diagnosis will save the child from unnecessary work-up and kidney involvement. PMID- 12751002 TI - Sonographic biometry of liver and spleen size long after closure of abdominal wall defects. AB - Little is known about the fate of the liver and spleen after closure of the abdominal cavity in patients with abdominal wall defects (AWD). Therefore, counselling families for long-term follow-up and in the case of surgery for acute disease, pregnancy or trauma may be difficult. A total of 18 patients ranging in age from 7 to 18 years, with AWD closed at birth, underwent ultrasound evaluation of liver and spleen size by determination of the index of liver size (ILS) and splenic volume (SV). These values were then correlated with some anthropometric parameters such as body mass index (BMI) and weight; correlation was also sought with some clinical features such as type of defect and direct or staged closure. Nearly all subjects exhibited weight above and BMI below the 50th percentile for age. ILS and SV were significantly above normal limits in all cases and no difference was found with regard to the type of defect. CONCLUSION: In patients having undergone surgery for abdominal wall defects, liver and spleen usually regain their normal shape and position even though size and volume appear to be larger than in normal controls. PMID- 12751003 TI - An 11-month-old boy with psychomotor regression and auto-aggressive behaviour. PMID- 12751004 TI - [Social status and health of young people in the German Federal State of Brandenburg]. AB - In the German Federal State of Brandenburg, reporting on health and social issues is closely linked. This also applies to the present article, which attempts to describe the social and health situation of young people in Brandenburg. The aim is to define future priorities in health prevention. METHODS: In addition to data from official statistics (on demographic trends, employment and social security) and from surveys conducted among young adults, the results from serial medical and dental examinations of 10th-grade pupils are presented. RESULTS: The number of young citizens in Brandenburg aged 16-25 will rise until the middle of the decade but then fall sharply. This will apply in particular to the rural areas far from Berlin. Sociological studies show that "work and family" enjoy the highest priority among people. However, less educated young people are increasingly doubtful about their ability to live their lives according to these priorities. According to medical statistics, every tenth young person suffers from allergies (especially allergic rhinitis). The same applies with regard to orthopaedic data. Approx. 5% of young people are obese, pupils of special schools more frequently than grammar school pupils. DISCUSSION: Healthy schooling, healthy professional training and good opportunities on the labour market are the key pillars of health policy in Brandenburg. The social and health situation of young people in Brandenburg is a complex issue and can only be improved by networking and cooperation among of all parties concerned. PMID- 12751005 TI - [Mental health promotion and prevention of mental diseases: are there population based concepts? Looking abroad]. AB - BACKGROUND: The rising public health relevance of mental health requires increased activities in the field of mental health promotion and prevention. In recent times interest in this sphere increases in Germany. OBJECTIVE: Which kinds of population-based mental health promotion programmes and prevention approaches are internationally available? What can be learned from their example? METHOD: Systematic search of the Internet and literature databases. Analysis and comparison of the available mental health promotion and prevention concepts with regard to targets, sub-targets, target groups, protagonists, settings, outcome indicators and evidence management. RESULTS: Concepts for mental health promotion and prevention programmes exist in England, Australia, the European Union and in Quebec, Canada. The concepts show similarities such as choosing similar settings like work place and schools or similar target groups like caretakers or adults in particular challenging life circumstances. CONCLUSIONS: Concepts for population based mental health promotion and prevention have been created in some countries. For developing such programmes in Germany one should refer to internationally available experience as well as to own gathered experiences as to own expressions gathered pilot projects and the German Health Target Programme. PMID- 12751006 TI - [Do local nursing home inmates get appropriate medication?--comments on the quality of drug application in nursing homes]. AB - BACKGROUND: Preparing prescribed drugs in unit dose systems for often multimorbid patients in nursing homes is a responsible and difficult task for the nursing personnel. The aim of this study was to increase the knowledge on the quality level of drug supply. METHOD: In a non-representative sample the local inspectorates for pharmacies in North Rhine-Westphalia compared the actually available drugs with the prescribed drugs documented in nursing homes. Participation was voluntary. RESULTS: In 50 % of the investigated nursing homes no medication error could be found. 30 percent of the observed cases showed only one error. In 20 percent the individual preparation of drugs in unit dose systems should be improved. Nursing homes with specific quality standards showed the least number of errors. Oral and not documented arrangements seem to be a common cause for avoidable errors. PMID- 12751007 TI - [New research avenues in exploring causes of death in Germany via regional statistical offices as exemplified by a retrospective cohort study]. AB - In retrospective epidemiological cohort studies the mortality rate of a cohort exposed to certain agents under investigation is compared to the mortality rate of the general population. In Germany, vital status of the cohort is followed up via local population registries. Cause of death of deceased members of the cohort is obtained from death certificates by local health authorities. Since storage time for death certificates is short in some Federal German states, causes of death cannot be obtained if death occurred before storage deadline. Recently it became possible to obtain the cause of death from regional statistical offices. Initial co-operation has been established with the statistical offices of North Rhine Westphalia and Berlin. For a cohort study in the chemical industry, the initially low proportion of causes of death obtained from local health authorities (44.8 %) was increased by 30 % to 74.0 % after obtaining ICD coded causes of death from two regional statistical offices. Further advantages of this new data source are reduction in duration of inquiry time and of overall cost. PMID- 12751008 TI - [Diffusion of the non-smoking campaign "Be Smart-Don't Start" between 1997 and 2003 in Germany]. AB - BACKGROUND: In Germany there is a high prevalence of smoking especially among young people. A comprehensive tobacco control programme combining several single measures is necessary. One of these measures should be health education programmes, including programmes for primary prevention of smoking. One recently developed primary smoking prevention programme is the competition "Be Smart - Don't Start". This programme has been implemented in Germany since the school year 1997/1998. The aim of this study is to describe the diffusion of the programme between 1997 and 2003. METHODS: The diffusion of the programme over a six-year period in Germany is described. School classes were clustered according to grade and region (German "Bundeslander"). The estimated number of participating pupils is compared to population data. RESULTS: Between 1997 and 2003 the number of participating classes has grown by the factor 18.2: from 462 school classes in the school year 1997/1998 to 8,402 school classes in the school year 2002/2003. In the school year 2002/2003, 218,452 pupils participated in the competition, which equals 5.64 % of the total population of the 11- to 14-year olds. The number of pupils participating in the programme varies to a high degree in Germany. Between 1.07 % and 12.2 % of the target group is reached via the programme. In some grades and regions of Germany up to 19 % of the target group participate in the competition. CONCLUSION: The competition "Be Smart - Don't Start" has been successfully implemented in Germany. To our present knowledge the programme is the biggest primary smoking prevention programme in Germany. PMID- 12751009 TI - [Hygienic quality of the water of public swimming pools--comments on a current German ordinance draft]. AB - In April 2002, a new draft regarding the quality of the water of public swimming pools in Germany was published. This draft was adapted to the new regulations for drinking water in respect of its structure and regulations. Microbiological tests for assessing the quality of bathing water are given priority. With regard to basic differences in facilities for drinking water and such for bath water and to legal aspects, many data (n = 14,425 data) on the practical experience of public health authorities are presented, obtained during the last few years. With regard to the data and in respect of the great delay between sampling and obtaining the results it is concluded that microbiological methods are inappropriate parameters for rapid assessment of the quality of bath water. Hence it is recommended to implement the evaluation of disinfection by chlorination including pH as an indicator for the quality of a bath water, parallel to the guidelines on technical standards. A high frequency of chlorination testing can markedly reduce the number of microbiological tests. Additionally, microbiological tests of the filtrate before disinfection by chlorination are recommended. This procedure has proved successful in legionella control. PMID- 12751010 TI - [Malaria surveillance in Germany 2000/2001--results and experience with a new reporting system]. AB - In Germany, malaria is one of the most frequently imported infectious diseases. In January 2001, the newly implemented Infektionsschutzgesetz (Law for Protection Against Infectious Diseases) brought some important changes in surveillance case notification procedures. After one year, experience shows that the changes did not affect the continuity and representative nature of malaria reporting in Germany. In the years 2000 and 2001, 836 and 1,040 malaria cases, respectively, were reported. In both years, most of the patients were between 30 and 49 years old. 82 % of the infections had been acquired in Africa, and 11 % in Asia. The predominant parasite species was P. falciparum (70 %), followed by P. vivax (12 % in 2000, and 16 % in 2001, respectively). The majority of infections occurred among tourists, fewer in immigrants or business travellers. About two thirds of all patients had not taken any chemoprophylactics. Compared to previous years a decrease in the number of fatal cases was observed (1998: 21, 1999: 18, 2000: 8, 2001: 8). To ameliorate the situation of imported malaria intensified prevention activities are necessary, including efforts to improve choice and compliance of chemoprophylaxis and to facilitate timely diagnosis and treatment. PMID- 12751011 TI - [Nosocomial transfer of HBV and HCV by public health workers]. AB - Transmission of HBV and HCV from people who work in medical professions to their patients is still an unsolved hygienic and legal problem. In Germany, cases of nosocomial hepatitis virus infection in health care units have received great public interest. Medical examinations of the employees according to occupational safety regulations aim at the employees only. Legal regulations including regulations of the European Union limit the purpose of these examinations on safety and health of the employees. These examinations do not serve the safety of patients. Protection against infections is regulated by the relevant German public health law, however regulations--especially those that concern the protection of the public--are incomplete. In Germany it is mandatory to inform the public health departments only in cases of acute hepatitis. Doctors do not need to give information about chronic liver infections. This may lead to the situation that a health care worker is unaware of a chronic, potentially infectious condition and his immunological status may remain unknown for a long period. Examinations in occupational medicine cannot solve this problem. In order to improve the protection of the public, there is a need to extend the regulations concerning the notification of chronic hepatitis and to implement solutions for this difficult and sensible problem in Germany. PMID- 12751012 TI - [Penetrating atherosclerotic ulcer of the thoracic aorta descendens] AB - Penetrating atherosclerotic ulcer of the thoracic aorta descendens. HISTORY AND CLINICAL FINDINGS: A 75-year-old man with a history of generalised atherosclerosis was admitted to hospital for invasive assessment of progredient typical angina pectoris. Apart from diminished peripheral pulses, physical examination was normal. INVESTIGATIONS: Coronary angiography revealed a three vessel coronary artery disease. The chest X-ray showed elongation and dilatation of the distal aortic arch and the proximal descending aorta thoracalis. Computed tomography and magnetic resonance imaging of the thorax as well as magnetic resonance angiography of the thoracic aorta, demonstrated a penetrating atherosclerotic ulcer of the descending aorta thoracalis, with extensive intramural hematoma. TREATMENT AND COURSE: After percutaneous ballon-dilatation of the right coronary artery and the circumflex artery, the patient was asymptomatic. Considering all aspects of the patients condition, medical treatment of the penetrating atherosclerotic ulcer was decided for the patient. The findings of the thoracic computed tomography after 6 months were unchanged. CONCLUSION: The penetrating atherosclerotic ulcer of the thoracic aorta is a less known clinical entity. Our case report demonstrates that even extensive forms can be clinical asymptomatic and discovered by routine radiologic examinations. PMID- 12751013 TI - [Secondary splenic rupture after thrombolysis for acute myocardial infarction] AB - Secondary splenic rupture after thrombolysis for acute myocardial infarction. HISTORY AND ADMISSION FINDINGS: A 67-year-old male patient was admitted with acute chest pain and signs of an acute anterior myocardial infarction in the ECG. The usual contraindications were excluded and after a systemic lysis with rt-PA the ECG-alterations as well as the symptoms of angina resolved completely. 2 hours later the patient developed an acute abdomen with a severe circulatory shock. INVESTIGATIONS: On ultrasound and CT a massive intraabdominal bleeding was found. TREATMENT AND COURSE: Emergency laparotomy revealed a splenic rupture. Retrospectively, 6 weeks before admission, the patient had fallen from a ladder to his left side. This is a rare case of a secondary splenic rupture during thrombolysis for acute myocardial infarction. 2 weeks later the patient developed rein-farction with angiographically shown two vessel disease. After angioplasty of the ramus interventricularis anterior (RIVA) he was stable. CONCLUSIONS: Intravenous thrombolysis in case of acute myocardial infarction is the method of choice. In the past a great number of patients were excluded from thrombolysis because of an extensive interpretation of contraindications. The aim to reach an alteration in this use may not risk health of patients by insufficient history. PMID- 12751014 TI - [Ventricular septal defect following cardiac trauma: closure with the Amplatzer Septal Occluder] AB - Ventricular septal defect following cardiac trauma: percutaneous closure with the Amplatzer septal occluder. HISTORY AND CLINICAL FINDINGS: A 36-year old roofer fell 8 m and suffered a severe polytrauma. A complicating pericardial tamponade was relieved as an emergency and myocardial fissure of the left ventricle about 1 cm in length sutured. 2 weeks later, a severe mitral insufficiency due to rupture of the papillary muscle occurred, that was cared by the implantation of a bioprosthesis. 6 weeks later dyspnoea and restricted physical capability were clinically impressive. INVESTIGATIONS AND DIAGNOSIS: Echocardiography demonstrated a posttraumatic muscular ventricular septal defect. Doppler echocardiography and heart catheterization showed a ventricular septal defect still restrictive with a left-to-right shunt (pulmonary-to-systemic flow ratio Qp/Qs 1.8:1). Under exercise, there was a significant increase in mean pulmonary arterial pressure from 27 to 60 mmHg. TREATMENT AND COURSE: The patient who had already been operated twice before was treated by the percutaneous occlusion of the ventricular septal defect from arterio- to venofemoral, a guide catheter was inserted transseptally into the left ventricle. An Amplatzer Septal Occluder, a self-expandable, self-centering wire-mesh double disc with a connecting central stent part, was loaded and then implanted in the ventricular septal defect. The intervention was controlled by fluoroscopy and echocardiography. Post intervention, only a trivial residual shunt was seen. The pumping efficacy of the left ventricle increased, in particular of the septal and apical segments. Clinically, the patient was markedly more load-bearing, the exercise-induced dyspnoea reduced. CONCLUSIONS: Following a cardiac trauma, various complications may occur that can manifest themselves clinically at two or more times. A posttraumatic ventricular septal defect of a patient already operated was successfully occluded by an Amplatzer Septal Occluder. Alongside established surgical methods, the non-operative implantation of new occlusion systems could mean an effective treatment option for muscular ventricular septal defects. PMID- 12751015 TI - [67-year-old patient with upper gastrointestinal bleeding] AB - A 67-year-old patient with upper gastrointestinal bleeding. HISTORY AND CLINICAL FINDINGS: A 67-year old patient with melaena and anaemia (haemoglobin 4.8 g/dl) for 14 days had been admitted to hospital under suspicion of upper gastrointestinal bleeding. The patient had a history of 2/3 resection of the stomach for recurrent duodenal ulcer and of a right nephrectomie for hypernephroma 18 years ago. There was no abdominal pain on pressure, nor muscular guarding. INVESTIGATIONS: Endoscopy (oesophago-duodenoscopie, coloscopie), radiography (angiography, gastrografin passage) and ultrasound failed to locate the source of bleeding. TREATMENT AND COURSE: After repeated endoscopic attempts to localize the source of bleeding and growing need of transfusions (12 blood concentrates in 5 days) an exploratory laparotomie was done. Surprisingly a bleeding fistula between the stump of the right renal arteria and duodenum was found. The defects were excised and directly closed. CONCLUSIONS: Secondary aortoduodenal fistula after nephrectomy is rare. If the source of bleeding can't be found, and there is in any doubt an exploratory laparotomy should be done. PMID- 12751016 TI - [Hypothyreoidism with thyroglobulin antibodies during corticoid replacement in a 54-year-old man with isolated ACTH deficiency] AB - Hypothyroidism with thyroglobulin antibodies during corticoid replacement in a 54 year-old man with isolated ACTH deficiency. HISTORY AND ADMISSION FINDINGS: A 54 year-old previously healthy man was admitted because of fatigue, tiredness, diarrhoea and weight loss for the last 3 years. Physical examination revealed dry but normally pigmented skin and markedly reduced Achilles reflex bilaterally. INVESTIGATIONS: Erythrocyte sedimentation rate was slightly elevated at 32 mm/h, C-reactive protein was normal. Both haemoglobin (12.4 mg/dl) and the corpuscular indices were normal, as were serum electrolytes, and sodium bicarbonate. But basal levels of thyroid stimulating hormone (TSH, 8.5 mU/ml) was markedly elevated, while free peripheral triiodothyronine (3.2pg/ml) was normal and free thyroxine (fT4) at 0.7 ng/d was slightly reduced. Thyroid ultrasound was normal. Test for antinuclear antibodies was slightly positive, but double-strand DNA was not demonstrated. Antithyroglobulin antibodies were slightly raised to 1012 IU/ml (normal <350). The basic level of ACTH was repeatedly below detection, as were plasma cortisol and cortisol excretion in 24-hour urine. Nuclear magnetic imaging was normal. Failure to stimulate corticol synthesis in the short ACTH test and by CRH indicated an isolated ACTH deficiency at the level of the anterior pituitary, while other hypophyseal functions were unaffected. TREATMENT AND COURSE: The patient"s condition rapidly improved on replacement with hydrocortisone, 30 mg/d, and thyroxine, 100 mg/d. No thyroglobulin antibodies or antinuclear antibodies were demonstrable after 6 months. Thyroxine was discontinued after 15 months. Frequent monitoring of thyroid function over the next 2 years always indicated a euthyroid state. CONCLUSION: Subnormal concentration of peripheral thyroid hormone combined with elevated TSH levels can, in the presence of hypercorticolism, be due to reversible abnormal thyroid function. PMID- 12751017 TI - [Unclear pulmonary infiltrates with eosinophilia, a problem of differential diagnosis] AB - Unclear pulmonary infiltrates with eosinophilia, a problem of differential diagnosis. HISTORY AND ADMISSION FINDINGS: A 60-year-old woman was admitted for the diagnosis of pulmonary infiltrates. A year before she had been exposed to tuberculosis when working as a doctor in Manila, the Philippines. Ten days before admission she had spent 10 days in Sao Paulo, Brazil. On admission she complained of fatigue, dry cough and nocturnal sweating. Her body temperature was 37.8; C. At auscultation of the chest fine rales were heard with diminished percussion sounds over both lungs. INVESTIGATIONS: The chest radiogram showed bilateral apical infiltrates. Blood count indicated normal white and red cells, but platelets were raised to 606 x 10 9/l. The differential blood count revealed an eosinophilia of 30%, ESR was raised at 91 mm/h and C-reactive protein increased to 103 mg/l. Angiotensin-converting enzyme, IgG, IgA, IgM, IgE, C3 and C4, paraproteins, antinuclear antibodies and double-strand DNA antibodies were all within normal limits. There was no direct or indirect evidence of tuberculosis and no parasites were found in sputum, stool, urine and blood. DIAGNOSIS, TREATMENT AND COURSE: After bronchoscopy with bronchial biopsy had failed to establish a diagnosis, an open lung biopsy with partial lung resection was performed. This revealed histologically an eosinophilic pneumonia with intra alveolar protein precipitation and multinucleated giant cells, as well as interstitial fibroblast proliferation without demonstrable mincroorganisms. Under cortisone administration there was striking improvement of symptoms within a few days, and C-reactive proteins fell to 3 mg/l, ESR to 25 mm/h and the eosino philia to 2%. CONCLUSION: Eosinophilic pneumonia should be included in the differential diagnosis of unclear pulmonary infiltrations with eosinophilia, once parasitological and malignant diseases, tuberculosis and allergic pulmonary aspergillosis have been excluded. PMID- 12751018 TI - [Thalidomide as a new treatment option in patients with multiple myeloma] AB - Thalidomide as a new treatment option in patients with multiple myeloma. HISTORYAND ADMISSION FINDINGS: In a 56 year old woman the diagnosis of multiple myeloma of subtype IgG kappa led to the performance of an autologous stem cell transplantation. After initial partial remission the disease became refractory to standard chemotherapy requiring repeated plasmaphereses due to hyperproteinemia. INVESTIGATIONS: Before application of thalidomide the patient had a serum protein of 12 g/dl and a paraprotein of 4.4 g/dl. Bone marrow histology revealed a 40 % infiltration of the marrow by differentiated plasma cells. On neurologic examination a slightly diminished nerve conduction speed of the right median nerve due to carpal tunnel syndrome was diagnosed. TREATMENT AND COURSE: Treatment with thalidomide in a dosage of 400 mg p. o. was initiated and led to a partial remission with a > 50 % reduction of the paraprotein within four weeks. Due to a neuropathy (WHO grade 1) a dose reduction to 200 mg per day was performed after seven weeks of treatment. 48 weeks after initiation of treatment the patient remains in a stable partial remission with a paraprotein of 1.4 g/dl. After 20 weeks of treatment 5 % of the bone marrow remained infiltrated by monoclonal plasma cells. CONCLUSION: Monotherapy with thalidomide led to a stable partial remission in a patient with refractory multiple myeloma. In vitro thalidomide is a potent inhibitor of angiogenesis. Besides, a reduction of TNFalpha as well as an inhibition of cytokines have been described. The exact mechanism of the drug in multiple myeloma remains elusive. However several phase II trials have demonstrated a beneficial effect in relapsed or refractory disease. The optimal dosage and timing have to be studied in further trials. PMID- 12751019 TI - [Neuropathy after cyclophosphamide high dose chemotherapy in a Morbus Werlhof patient] AB - Neuropathy after cyclophosphamide high dose chemotherapy in a Morbus Werlhof patient. PERSONAL HISTORY: A 24 year old patient with longstanding autoimmune idiopathic thrombocytopenic purpura (M. Werlhof) was treated with glucocorticoids, immunoglobulins, splenectomy, immunosuppression and vincristin without lasting success. After a second treatment cycle with cyclosphosphamide and autologous peripheral stem cell transplantation she acutely develpod symptoms of a peripheral sensoric and motoric polyneuro-pathy. MEDICAL EXAMINATION: At admission she was in good general health, but had steroid-induced Cushing's symptoms, generalized petechial bleeding and thrombocytopenia (1000/ micro l). THERAPY AND COURSE: After 2.5 g/m 2 cyclophosphamide and stem cell transplantation distally pronounced polyneuropathy developed within a week with bladder insufficiency. Major bleeding or brain damage were excluded, and symptoms only partially reversed when treated with steroids, carbamazepine and amitriptyline. Thrombocytopenia persisted, and the patient died 4 month later from acute brain hemorrhage. CONCLUSIONS: Direct neurotoxicity has to be assumed as the likely causative agent in this case, illustrating the possibility of peripheral neuropathic lesions by high dose cyclophosphamide treatment. PMID- 12751020 TI - [Severe Vitamin D(dihydrotachysterol)-intoxication with temporary anemia and hypercalcemia responsive to bisphosphonates] AB - Severe Vitamin D(dihydrotachysterol)-intoxication with temporary anemia and hypercalcemia responsive to bisphosphonates. HISTORY AND FINDINGS: A 31-year old female patient presented with pain of her skeletal system. 6 months prior to her presentation, she underwent thyroid surgery for Graves disease. She also suffered from endocrine orbitopathy. Afterwards, she expirienced surgical hypoparathyroidism and received dihydrotachy-sterol (A.T.10 (R)) in a dose of up to 4 mg per day. The physical examination was unremarkable except for the presence of Graves' ophthalmopathy and a swelling at the left ancle. INVESTIGATIONS: Upon admittance, she had severe hypercalcemia (serum calcium: 4.1 mmol/l) with plasma intact PTH levels below the limit of detection, renal failure (serum creatinine: 5.5 mg/dl) and normocytic anemia (initial hemoglobin: 8.3 g/dl, upon rehydration: 6.6 g/dl). TREATMENT AND COURSE: Upon rehydration and induced diuresis, the renal function improved and the serum calcium came down rapidly in the early treatment phase. However, serum calcium remained elevated at around 3.0 mmol/l after 4 weeks. Only after the use of the bisphosphonate pamidronate (15 mg), serum calcium returned into the normal range and remained there. Treatment for hypoparathyroidism had to be reinstituted only 20 weeks after dihydrotachysterol had been discontinued. The anemia had resolved without any treatment at that time. CONCLUSIONS: Treatment with dihydrotachysterol and other long-acting Vitamin D preparations has to be monitored closely because of the risk of severe hypercalcemia, which may be difficult to treat. In our case, dihydrotachysterol was still active until week 20 after the drug was discontinued. Anemia should be considered as a side effect of dihydrotachysterol intoxication and does not warrant elaborate differential diagnosis in this context. A single administration of a bisphosphonate turned out to be of major therapeutic benefit and resulted in a lasting correction of hypercalcemia. Therefore, bisphosphonates should become part of the treatment regimen for vitamin D intoxication. PMID- 12751021 TI - [Bodily symptoms and utilization of medical care] AB - Bodily symptoms and utilization of medical care. OBJECTIVE: An important aspect in a person's attitude towards disease, which also determines when medical help is sought, relates to one's perception of bodily symptoms and to the manner in which the impact of an illness is dealt with. It was the aim of this study to ascertain what bodily signs of disease in an organ system would make healthy persons within a normal population seek medical help. MATERIAL AND METHODS: A representative sample of the population of Germany (n=2050), aged between 14 and 92 years, was presented with the description of 20 symptoms of diseases of various organ systems and asked which of these symptoms would induce them to consult a doctor. RESULTS: Between 8% and 10% of those asked said that they would not go to a doctor even if they had clear bodily symptoms, e. g. blood in the urine , persistent joint pains , dizziness of fainting or pain in the lower abdomen . Analysis of variance for individual symptoms revealed that a significantly higher percentage of women and elderly persons would go and see a doctor than men and younger persons. Significantly fewer persons in the eastern (former DDR) Lands than the western ones would seek medical help. There was no significant correlation between the seeking medical help and subjective complaints, as measured by the Giessen Complaint Questionnaire, as well as any tendency towards hypochondria, as measured by the Whiteley Hypochondria Index. CONCLUSION: It is clear that complex processes of bodily awareness and assessment of symptoms precede medical consultation. These processes must be considered as part of a person's attitude towards illness and, most of all, of its congnitive component ( health beliefs ). They are dependent on charakteristics of sociodemography and social structure ant they have far reaching significance for ambulatory care. It is clear that there is a great need for research in this area. PMID- 12751022 TI - [Myocardial infarction and coronary artery ventricular fistulas due to blunt chest trauma] AB - Myocardial infarction and coronary artery ventricular fistulas due to blunt chest trauma. HISTORY AND CLINICAL FINDINGS: An 18-year-old previously healthy, cigarette smoking man with no other risk factors for ischaemic heart disease, was admitted to hospital after being kicked in the chest by a horse. On arrival he complained about pain in the lower mediastinum. INVESTIGATIONS: The ECG showed sinus rhythm, right bundle branch block and convex bowed ST elevation in leads V1 V3. Sixty minutes after the incident the cardiac enzymes (creatinekinase-MB fraction, troponin I) were significantly raised. Despite an only slightly reduced left ventricular function documented by transthoracic echocardiography, SPECT thallium scan showed large scintigraphic defects. Coronary heart disease was excluded by coronary angiography. Four small coronary-ventricular fistulas were identified. Laevocardiography showed a hypokinesia in the antero-septal region. DIAGNOSIS, TREATMENT AND COURSE: We assumed traumatic myocardial infarction of the anterior wall and rupture of multiple small coronary vessels, leading to coronary-ventricular fistulas. No interventional or surgical therapy was performed. Later on the left ventricular function became normal. Echocardiography merely outlined an akinetic scar in the middle of the septum. At exercise ECG test sixteen months later, the patient remained asymptomatic and was able to exercise without any signs of ischaemia up to a work load of 175 W. Furthermore, the fistulas could be seen by echocardiography. CONCLUSION: Cardiac involvement should be considered in all cases of blunt chest trauma. In addition to a traumatic myocardial infarction fistulas may also, though rarely, occur. Myocardial scintigraphy after cardiac contusion is not suitable for diagnosing myocardial ischaemia or vitability. PMID- 12751023 TI - [Symptomatic cyst of the pancreas and asymptomatic bilateral phaeochromocytoma] AB - Symptomatic cyst of the pancreas and asymptomatic bilateral phaeochromocytoma. HISTORY AND CLINICAL FINDINGS: A 39-year-old woman was admitted to our department of gastroenterology with recurrent epigastrical pain. Ten years previously the diagnosis of von Hippel-Lindau (VHL)-syndrome has been established. Two years before a germ line mutation in exon 3 of the VHL-tumour suppressor gene has been detected. The patient has a healthy son with a normal VHL-gene and four healthy siblings who had refused a genetic blood test. INVESTIGATIONS: At abdominal ultrasound at the head of the pancreas three 4 2 cm large cysts and in the region of the left adrenal gland a 2,9 2,7 cm large tumor were visible. MRI of the abdomen revealed in addition a 2,2 1,5 cm large tumour of the right adrenal gland. An asymptomatic biadrenal phaeochromocytoma was detected by elevated urine catecholamines and 123I-MIBG-scintigraphy. TREATMENT AND COURSE: The cysts of the pancreas were punctured under endosonographical control and analysis of the cyst fluid was not suspicious of a malignant cystic tumour. The patient had no further abdominal complaints. After oral treatment with the alpha-blocker phentolamine the biadrenal phaeochromocytoma was treated by retroperitoneal laparoscopic surgery in an organ-sparing fashion. Postoperatively ACTH-stimulating test revealed a normal cortisol response. CONCLUSION: Adrenocortical function can be preserved by la-paroscopic adrenal-sparing surgery in bilateral phaeochromocytoma. PMID- 12751024 TI - Heritability analysis of cytokines as intermediate phenotypes of tuberculosis. AB - Numerous studies have provided support for genetic susceptibility to tuberculosis (TB); however, heterogeneity in disease expression has hampered previous genetic studies. The purpose of this work was to investigate possible intermediate phenotypes for TB. A set of cytokine profiles, including antigen-stimulated whole blood assays for interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta, and the ratio of IFN to TNF, were analyzed in 177 pedigrees from a community in Uganda with a high prevalence of TB. The heritability of these variables was estimated after adjustment for covariates, and TNF-alpha, in particular, had an estimated heritability of 68%. A principal component analysis of IFN-gamma, TNF-alpha, and TGF-beta reflected the immunologic model of TB. In this analysis, the first component explained >38% of the variation in the data. This analysis illustrates the value of such intermediate phenotypes in mapping susceptibility loci for TB and demonstrates that this area deserves further research. PMID- 12751025 TI - Role of leukocyte immunoglobuin G receptors in vaccine-induced immunity to Streptococcus pneumoniae. AB - Members of the leukocyte immunoglobulin (Ig) G receptor (FcgammaR) family play a key role in antibody-mediated phagocytosis and can either enhance antigen presentation or down-modulate immune responses. We studied immune responses to a pneumococcal conjugate (pneumococcal polysaccharide serotype 1 [PPS1]-tetanus toxoid) and antibody-mediated protection in mice deficient for individual FcgammaRs and complement receptor 3 (CR3). FcR gamma chain-deficient (FcR gamma chain(-/-)) mice, which lack expression of both FcgammaRI and III, had significantly lower anti-PPS1 IgG2b and IgG3 responses than did wild-type mice, whereas FcgammaRII-deficient (FcgammaRII(-/-)) mice had significantly higher IgG2a and IgG3 titers. Wild-type and FcgammaRII(-/-) mice were protected against infection with pneumococcal serotype 1, whereas immunized FcR gamma chain(-/-) and FcgammaRIII-deficient mice were not. Immunized CR3-deficient mice were protected against disease, and complement depletion had little effect on protection. These data indicate that activatory leukocyte FcgammaR, but not FcgammaRII (a murine homologue of human FcgammaRIIb), contributes to IgG-mediated protection against pneumococcal disease. PMID- 12751027 TI - Functional antibodies elicited by an 11-valent diphtheria-tetanus toxoid conjugated pneumococcal vaccine. AB - The aluminium-adjuvanted 11-valent pneumococcal conjugate vaccine containing polysaccharides 1, 4, 5, 7F, 9V, 19F, and 23F (coupled to tetanus protein) and polysaccharides 3, 6B, 14, and 18C (coupled to diphtheria toxoid) elicits high antibody concentrations in Filipino infants when given at ages 6, 10, and 14 weeks and 9 months simultaneously with the national vaccination program. We evaluated functional activity of these antibodies by using a viable cell opsonophagocytic assay (OPA). The OPA titers correlated (r=0.53-0.74) with the respective antibody concentrations. The geometric mean OPA titers after 3 vaccine doses were 276.9 (serotype 4), 12.3 (serotype 6B), 46.0 (serotype 14), 119.3 (serotype 19F), and 206.3 (serotype 23F). The functionality of antibodies increased after the fourth dose of vaccine (i.e., the concentration required for in vitro killing of pneumococci decreased). Both the quantity and quality of antibodies are important in the evaluation of immunogenicity of pneumococcal vaccines. PMID- 12751026 TI - Contribution of protein G-related alpha2-macroglobulin-binding protein to bacterial virulence in a mouse skin model of group A streptococcal infection. AB - Protein G-related alpha(2)-macroglobulin-binding (GRAB) protein is a cell wall attached determinant of group A streptococcus (GAS) that interacts with the human protease inhibitor alpha(2)-macroglobulin (alpha(2)-M). Of 86 clinical isolates tested, 23% could bind alpha(2)-M. However, all strains tested contained the grab gene. High levels of anti-GRAB antibodies were found in the serum of convalescent GAS-infected patients, a finding that indicates that this protein is expressed during the infection process. Among the alpha(2)-M-binding strains, 80% were skin isolates, and 20% were throat isolates, findings that suggest that the skin environment is a preferential site for expression of alpha(2)-M-binding activity. To test this possibility, we determined the role of GRAB in a mouse model of GAS skin infection. The wild-type strain KTL3, which interacts with alpha(2)-M, showed high virulence. The isogenic mutant of KTL3, MR4, devoid of surface-bound GRAB, was attenuated in virulence, compared with the wild-type strain. Thus, mice infected with MR4 survived longer, developed smaller skin lesions, and exhibited lower levels of bacterial dissemination than did those infected with KTL3. These results emphasize the role of GRAB as a virulence factor of GAS. PMID- 12751028 TI - High rate of macrolide resistance in Staphylococcus aureus strains from patients with cystic fibrosis reveals high proportions of hypermutable strains. AB - Incidence of resistance to erythromycin at our institution reached 53% in 122 Staphylococcus aureus isolates obtained from patients with cystic fibrosis (CF) from 1997 to 1999. Macrolide-resistance genes were sought for in 20 erythromycin resistant isolates from 9 patients with CF by use of polymerase chain reaction; 13 strains did not contain any known macrolide-resistance genes. Sequence of ribosomal genes rrl (23S rRNA), rplD (L4 protein), and rplV (L22 protein) revealed the presence of mutations in the target site of macrolides in 15 of the 20 isolates. A higher proportion of hypermutator strains was observed in a group of 89 CF staphylococcal isolates, compared with that in the 74 non-CF control isolates (13/89 vs. 1/74 with resistance to rifampin [P=.0045]; 9/89 vs. 1/74 with resistance to streptomycin [P=.04]). Various mutations or deletions of the mutator mutS gene were found not only in 5 of 11 hypermutable strains but also in 3 nonhypermutable strains harboring a large number of ribosomal mutations. The presence of a high proportion of hypermutable strains might explain the adaptation of certain strains in the patients, as well as the emergence of macrolide resistance as a result of antibiotic selective pressure in CF. PMID- 12751029 TI - Optical monitoring and treatment of potentially lethal wound infections in vivo. AB - We report on the use of optical techniques to monitor and treat Pseudomonas aeruginosa wound infections in mice. Bioluminescent bacteria transduced with a plasmid containing a bacterial lux gene operon allow the infection in excisional mouse wounds to be imaged by use of a sensitive charge-coupled device camera. Photodynamic therapy (PDT) targeted bacteria, by use of a polycationic photosensitizer conjugate, which is designed to penetrate the gram-negative cell wall and was topically applied to the wound and was followed by red-light illumination. There was a rapid light dose-dependent loss of luminescence, as measured by image analysis, in the wounds treated with conjugate and light, a loss that was not seen in untreated wounds, wounds treated with light alone, or wounds treated with conjugate alone. P. aeruginosa was invasive in our mouse model, and all 3 groups of control mice died within 5 days; in contrast, 90% of PDT-treated mice survived. PDT-treated wounds healed significantly faster than did silver nitrate-treated wounds, and this was not due to either inhibition of healing by silver nitrate or stimulation of healing by PDT. PMID- 12751030 TI - A low level of CD4+CD28+ T cells is an independent predictor of high mortality in human immunodeficiency virus type 1-infected patients. AB - This study investigated coexpression of CD28, CD45RA, and CD45RO on CD4(+) and CD8(+) cells in 107 human immunodeficiency virus (HIV) type 1-infected patients, who were followed-up prospectively and were not treated with highly active antiretroviral therapy, and 65 control subjects. The most important novel finding was that a 50% reduction in CD4(+)CD28(+) cells predicted increased mortality (relative hazards [HR], 1.6; 95% confidence interval [CI], 1.0-2.6; P=.04), even after adjusting for the CD4(+) cell counts, virus load, beta(2)-microglobulin and hemoglobin levels, and HIV disease stage. Patients with progressed HIV infection had decreased concentrations of all studied cell subsets. Concerning the proportions of cells, only CD4(+)CD28(+), CD4(+)CD45RA(+), and CD8(+)CD45RO(+) cells decreased with HIV progression. Low proportions of CD4(+)CD45RA(+), CD8(+)CD45RA(+), and CD8(+)CD45RO(+) cells predicted mortality only in univariate but not in multivariate Cox analyses. If our results are confirmed in other studies, coexpression of CD28 on CD4(+) cells may be a useful marker to evaluate HIV progression. PMID- 12751032 TI - Latent infection as a source of disseminated disease caused by organisms of the Mycobacterium avium complex in simian immunodeficiency virus-infected rhesus macaques. AB - Whether infection with Mycobacterium avium complex (MAC) among patients with acquired immune deficiency syndrome results from recent exposure to virulent strains or reactivation of latent infection acquired years earlier is unknown. To address this question, tissue samples from 47 simian immunodeficiency virus (SIV) infected and 63 SIV-uninfected rhesus macaques were cultured. MAC was cultured from 14 SIV-uninfected macaques (22.2%) and 32 SIV-infected macaques (68.1%); median bacterial burdens were 33.3 and 998.7 cfu/g, respectively. Genetically distinct strains of MAC were identified for 13 SIV-uninfected macaques (20.6%) and 15 SIV-infected macaques (31.9%). A genetically identical MAC strain (K128A) was identified for 25 SIV-infected macaques (53.2%) and 1 SIV-uninfected macaque (1.6%). Multivariate analysis identified infection with SIV/Delta(B670), diagnosis of an SIV-related tumor or opportunistic infection, and birth on site as risks for MAC infection. SIV-uninfected and SIV-infected macaques yielding unique strains of MAC were considered to have latent and reactivation infection, respectively, whereas animals infected with strain K128A were considered to have recent infection, demonstrating that both mechanisms occur among rhesus macaques. PMID- 12751031 TI - Polyclonal proliferation and apoptosis of CCR5+ T lymphocytes during primary human immunodeficiency virus type 1 infection: regulation by interleukin (IL)-2, IL-15, and Bcl-2. AB - We measured apoptosis of subsets of T lymphocytes by single-cell analysis of caspase activation, to confirm high turnover of chemokine receptor CCR5(+) T cells in subjects with acute, primary human immunodeficiency virus type 1 (HIV-1) infection (PHI). High levels of spontaneous apoptosis, consisting mainly of CD8(+) T lymphocytes, were closely associated with increases in the activation markers Ki-67, CD38, and the HIV coreceptor CCR5 and with decreases in Bcl-2 and the interleukin (IL)-7 receptor at the single-cell level. Increased expression of Ki-67 and CCR5 ex vivo, as well as increased apoptosis, was seen in all T cell receptor beta-chain variable region (TCRBV) subfamilies studied. The addition of IL-2 or IL-15, but not IL-7, significantly inhibited caspase activation, increased Bcl-2 expression, and rapidly initiated proliferation in vitro of CD8(+) T cells expressing CCR5 and multiple TCRBV subfamilies. Furthermore, IL-15 receptor alpha-chain messenger RNA levels were increased in peripheral blood mononuclear cells during PHI. These results suggest that CCR5(+)Ki-67(+)Bcl 2(dim) activated T cells generated during PHI traffic via blood to tissue sites, where the cells may survive and/or further proliferate under the local influence of IL-2 or IL-15. Understanding cytokine effects on CCR5(+) T cells will be important in understanding chronic HIV-1 replication and pathogenesis. PMID- 12751033 TI - The effects of varying exposure to malaria transmission on development of antimalarial antibody responses in preschool children. XVI. Asembo Bay Cohort Project. AB - In areas of intense malaria transmission, malaria morbidity and mortality is highest in children 3-18 months old. Interventions that reduce malaria exposure early in life reduce morbidity but may also delay development of clinical immunity. We assessed the relationship between intensity of malaria exposure and development of antibody responses. Thirty-nine children were monitored monthly, from birth to > or =2.5 years old (1238 observations), and were divided into 3 exposure categories, on the basis of parasitemic episodes or entomological data. Children with low exposure during the first 2 years of life had higher subsequent levels of antibody to merozoite surface protein-1(19-kDa) (a marker of blood stage responses) by months 24-35 (P<.05). This inverse relationship decreased as children aged. There was no consistent relationship between exposure early in life and subsequent levels of antibody to circumsporozoite protein (a marker of sporozoite-stage responses). These data suggest that, in areas of intense malaria transmission, during the first 3 years of life, interventions that either reduce the number of asexual parasitemic episodes or lower entomological exposure do not delay the development of antibody responses to blood-stage malarial antigens. PMID- 12751034 TI - Burden of malaria during pregnancy in areas of stable and unstable transmission in Ethiopia during a nonepidemic year. AB - Little is known about the epidemiology of malaria during pregnancy in areas of unstable (epidemic-prone) transmission (UT) in sub-Saharan Africa. In cross sectional studies, peripheral malaria parasitemia was identified in 10.4% of women attending antenatal care clinics at 1 stable transmission (ST) site and in 1.8% of women at 3 UT sites; parasitemia was associated with anemia in both ST (relative risk [RR], 2.0; P<.001) and UT (RR, 4.4; P<.001) sites. Placental parasitemia was identified more frequently during deliveries at ST sites (12/185; 6.5%) than at UT sites (21/833; 2.5%; P=.006). Placental parasitemia was associated with low birth weight at the ST site (RR, 3.2; P=.01) and prematurity at ST (RR, 2.7; P=.04) and UT (RR, 3.9; P=.01) sites and with a 7-fold increased risk of stillbirths at UT sites. The effectiveness and efficiency in Ethiopia of standard preventive strategies used in high-transmission regions (such as intermittent preventive treatment) may require further evaluation; approaches such as insecticide-treated bednets and epidemic preparedness may be needed to prevent adverse pregnancy outcomes. PMID- 12751035 TI - Zileuton reduces respiratory illness and lung inflammation, during respiratory syncytial virus infection, in mice. AB - To determine the role of leukotrienes in a mouse model of respiratory syncytial virus (RSV) infection, we correlated the quantity of leukotrienes in lung lavage fluids of infected mice with respiratory rates and measures of outflow obstruction and then determined the effects of the leukotriene inhibitor zileuton on clinical features and lung function. Concentrations of leukotrienes were correlated with both increasing respiratory rates and the degree of prolongation of expiratory time. Administration of zileuton 1 day before infection and through day 5 after infection markedly reversed airway constriction, reduced numbers of inflammatory cells in the lung, and prevented the weight loss associated with infection. Leukotrienes appear to contribute substantially to the pathogenesis of RSV-related disease. PMID- 12751037 TI - Rotavirus nonstructural protein NSP4 induces heterotypic antibody responses during natural infection in children. AB - Seroconversion of immunoglobulin A (IgA) and immunoglobulin G (IgG) (> or =4-fold rise) to rotavirus nonstructural protein 4 (NSP4) was determined, by use of enzyme-linked immunosorbent assay with fusion proteins glutathione S-transferase (GST)-NSP4 from strains SA11 (A), 116E (B), and RRV (C), in 40 children with acute rotavirus gastroenteritis and in 30 with the same disease due to other pathogens. The IgG seroconversion rates in the rotavirus group were 67.5%, 70%, and 60% when recombinant (r) NSP4A, -B, and -C, respectively, were used as antigen in the assay, and, for rotavirus-uninfected children, rates were 10%, 13%, and 7%. IgA seroconversion occurred in 57%, 70%, and 50%, respectively, of children with rotavirus gastroenteritis; in rotavirus-uninfected children, 1 child each seroconverted to the different rNSP4s. Among 9 children infected with strain NSP4A, 7, 6, and 5 children showed IgG seroconversion, and, among 18 infected with NSP4A, -B, and -C, 16, 17, and 15, respectively, showed IgG seroconversion. Between NSP4A-infected and NSP4B-infected children, IgA responses were similar to IgG responses. In conclusion, significant NSP4-specific antibody response occurs in natural rotavirus infection, and the antibody response appears to be broad and heterotypic in nature. PMID- 12751036 TI - Human herpesvirus 8 infection within families in rural Tanzania. AB - Human herpesvirus 8 (HHV-8) infection is common in Africa. We examined the distribution of HHV-8 within families in rural Tanzania to determine routes of spread. HHV-8 infection was assessed by measuring antibody reactivity with a K8.1 (lytic-phase antigen) immunoassay. The prevalence increased from 3.7% (1/27) among infants to 58.1% (36/62) among children aged 3-4 years and 89.0% (65/73) among adults aged > or =45 years. Women with HHV-8-seropositive husbands had a 7 fold risk for infection (odds ratio [OR], 6.9; 95% confidence interval [CI], 1.9 25.3). HHV-8 seropositivity in children was associated with having at least 1 seropositive first-degree relative (OR, 14.7; 95% CI, 5.9-43.1), a seropositive mother (OR, 7.4; 95% CI, 3.2-16.8), a seropositive father (OR, 4.8; 95% CI, 2.3 10.1), or a seropositive next-older sibling (OR, 4.2; 95% CI, 1.9-9.4). Our data are consistent with the occurrence of HHV-8 transmission within families, from mothers and other relatives to children via nonsexual routes and between spouses via sexual routes. PMID- 12751038 TI - Differential effects of priming with DNA vaccines encoding the hemagglutinin and/or fusion proteins on cytokine responses after measles virus challenge. AB - Measles is associated with a million deaths a year in developing countries because of secondary infections. Morbidity is particularly severe in young infants. Both measles-induced immune suppression and atypical measles have been associated with a type 2 cytokine bias of the immune response. The role of individual virus proteins in the induction of these cytokine responses is unknown and could be important for the development of new vaccines. We have used a rhesus macaque model and DNA vaccines to investigate cytokine responses to the individual measles virus (MV) protective antigens, hemagglutinin and fusion. The hemagglutinin protein primed for a type 2 cytokine response, with suppression of interleukin (IL)-12 and preferential production of IL-4 after MV challenge. The fusion protein primed for a type 1 response with preferential production of interferon-gamma. Responses were modulated when both proteins were used for priming. Therefore, the specific proteins included in a new measles vaccine will affect the type of cytokine response elicited. PMID- 12751039 TI - Dynamics of cytomegalovirus replication during preemptive therapy with oral ganciclovir. AB - The degree and dynamics of cytomegalovirus (CMV) replication were investigated in blood samples that were prospectively collected in the context of a placebo controlled study evaluating the efficacy of preemptive oral ganciclovir for the prevention of CMV disease after liver transplantation. The degree of viral replication was strongly associated with progression to CMV disease or viremia (risk ratio, 8.8 and 51.5 among patients with virus loads < or =2860 and >2860 copies/10(6) peripheral blood leukocytes, respectively). Preemptive oral ganciclovir therapy diminished the incidence of CMV disease or viremia but did not completely suppress higher levels of CMV replication. Six (21%) of 29 patients had persistent CMV replication during preemptive oral ganciclovir therapy; 2 patients subsequently developed "breakthrough" CMV syndrome. This study identifies a relative cutoff virus load that predicts subsequent development of CMV disease and highlights the inability of oral ganciclovir to suppress CMV replication in a subset of patients. PMID- 12751040 TI - Rhinovirus induction of the CXC chemokine epithelial-neutrophil activating peptide-78 in bronchial epithelium. AB - Epithelial-neutrophil activating peptide-78 (ENA-78) induces neutrophil migration, an early response to viral infection. Rhinovirus serotype 16 (RV16) was used to infect primary bronchial epithelial cells and a cell line (BEAS-2B). Release of ENA-78 protein was measured by enzyme-linked immunosorbent assay, ENA 78 mRNA production was quantified by reverse-transcription polymerase chain reaction, and ENA-78 promoter activity was assessed by use of a promoter construct. After infection with RV16, ENA-78 protein and mRNA increased significantly, and RV16 induced 3-fold increases in ENA-78 gene transcription. Nasal ENA-78 measured in patients with asthma with and without RV infection was more elevated in patients with RV infection present. Our study demonstrates that ENA-78 is produced in bronchial epithelial cells in response to RV16 infection. With other chemokines, it may be an important initiator of neutrophil airway inflammation during RV common colds and thus may play a role in the development of virus-associated airway pathologies. PMID- 12751041 TI - Human immunodeficiency virus-1 RNA levels in cerebrospinal fluid exhibit a set point in clinically stable patients not receiving antiretroviral therapy. AB - To characterize, over time, cerebrospinal fluid (CSF) virus-load change in clinically stable patients, human immunodeficiency virus (HIV)-1 RNA levels were measured in serial CSF samples and in plasma samples obtained, during periods of 20 days-6 years, from 17 HIV-infected individuals not receiving antiretroviral treatment. Longitudinal trends were analyzed by linear regression and restricted maximum-likelihood techniques. CSF HIV-1 RNA levels varied within a restricted range (+/-0.5 log(10) copies/mL) around each subject-specific mean. Although 16 of the 17 subjects had CSF slopes not significantly different from zero, slopes that were more positive were associated with lower CD4 counts. In an individual patient, a CSF virus-load change >0.5 log(10) copies/mL may be clinically significant. Furthermore, our data suggest that, if the CSF virus load reflects the size of the reservoir of infected cells in the central nervous system, this reservoir may increase in those individuals with advanced immunosuppression but is stable, over several years, in patients without AIDS. PMID- 12751042 TI - Impact of interleukin-1beta genetic polymorphisms on the development of hepatitis C virus-related hepatocellular carcinoma in Japan. AB - To examine the effects of polymorphisms in the gene encoding proinflammatory interleukin (IL)-1beta in patients infected with hepatitis C virus (HCV) in Japan, we studied 364 patients with chronic HCV infection (146 of whom had hepatocellular carcinoma [HCC] and 218 of whom did not) and 230 healthy control subjects. IL-1B-511 and IL-1RN genotypes were ascertained, and IL-1B-511 genotype T/T was found to be significant risk factors for the development of HCC, indicating that polymorphism in the IL-1B-511 genetic locus is one of the possible determinants of progression of hepatitis C to HCC. PMID- 12751043 TI - Human immunodeficiency virus (HIV) DNA load and level of immunosuppression in treatment-naive HIV-1-infected patients. PMID- 12751048 TI - On the evolving nature of understanding dialysis-related disorders. PMID- 12751044 TI - High prevalence of double Plasmodium falciparum dhfr mutations at codons 108 and 59 in the Sistan-Baluchistan province, Iran. PMID- 12751049 TI - The role of carnitine in normal and altered fatty acid metabolism. AB - Carnitine is a low-molecular-weight compound obtained from the diet that also is biosynthesized from the essential amino acids lysine and methionine. Carnitine has been identified in a variety of mammalian tissues and has an obligate role in the mitochondrial oxidation of long-chain fatty acids through the action of specialized acyltransferases. Other roles for carnitine include buffering of the acyl coenzyme A (CoA)-CoA ratio, branched-chain amino acid metabolism, removal of excess acyl groups, and peroxisomal fatty acid oxidation. The growing body of evidence about carnitine function has led to increased understanding and identification of disorders associated with altered carnitine metabolism. Disorders of fatty acid oxidation and metabolism typically are associated with primary and secondary forms of carnitine deficiency. These disorders, which include increased lipolysis, increased lipid peroxidation, accumulation of acylcarnitines, and altered membrane permeability, have significant consequences for patients with myocardial diseases and kidney failure. Therapeutic administration of carnitine shows promise in treating selected groups of patients who have altered carnitine homeostasis, resulting in improved cardiac function, increased exercise capacity, reduced muscle cramps, and reduced intradialytic complications. PMID- 12751050 TI - Dialysis-related carnitine disorder and levocarnitine pharmacology. AB - Among the homeostatic processes controlling the endogenous L-carnitine pool in humans, the kidney has a vital role through extensive and adaptive tubular reabsorption. Kidney disease can lead to disturbances in L-carnitine homeostasis, and long-term hemodialysis therapy can lead to a significant reduction in plasma and tissue L-carnitine levels and an increase in the ratio of acyl-L-carnitine to free L-carnitine. These alterations may interfere with the oxidation of fatty acids and removal from tissues of unwanted short-chain acyl groups. A dialysis related carnitine disorder (DCD) arises when these biochemical abnormalities exist in association with such clinical symptoms as muscle weakness, cardiomyopathy, intradialytic hypotension, or anemia that is resistant to erythropoietin therapy. Exogenous L-carnitine, administered intravenously, is approved for the treatment of secondary carnitine deficiency caused by long-term hemodialysis. Although intravenous administration of 20-mg/kg doses at the end of each hemodialysis session leads to supraphysiological levels of the compound in plasma, these levels do not appear to be associated with adverse effects. Because more than 99% of the body's carnitine pool is located outside of plasma, supraphysiological plasma levels appear to be required to ensure that depleted muscle stores can be replenished. Although oral L-carnitine has been used for the treatment of DCD, the bioavailability of oral L-carnitine is low (<15%) in healthy subjects and unknown in patients with end-stage renal disease. Moreover, gastrointestinal degradation of L-carnitine to trimethylamine and other compounds might limit the usefulness of long-term oral L-carnitine administration in this patient group. PMID- 12751051 TI - L-carnitine treatment of anemia. AB - Recombinant human erythropoietin (rHuEPO) and iron supplementation have had a profoundly positive impact on the anemia of patients with chronic kidney disease. However, a significant number of patients remain hyporesponsive to rHuEPO, with hemoglobin values less than target levels. A suboptimal response to rHuEPO is associated with complications that can reduce quality of life and increase morbidity, mortality, and costs. There are a number of other metabolic derangements associated with uremia that can impact on the production and survival of red blood cells. Dialysis-related carnitine disorder is a functional metabolic deficiency, common in chronic dialysis patients, that can have a negative impact on erythrocyte production and survival. This article reviews the role of L-carnitine in the pathogenesis and adjunctive treatment of anemia associated with kidney failure. After a comprehensive database search, primary and secondary reports were analyzed. Laboratory studies examining the influence of carnitine on red blood cell function and clinical trials of L-carnitine in dialysis patients support the use of L-carnitine in the setting of rHuEPO hyporesponsiveness. Consensus groups, including the National Kidney Foundation Kidney Disease Outcome Quality Initiative (K/DOQI), consider the use of L carnitine for hyporesponsive rHuEPO-dependent anemia a promising application of this therapy, recommending an empiric trial of L-carnitine in these patients. PMID- 12751052 TI - The role of carnitine in myocardial dysfunction. AB - L-Carnitine (carnitine) may have a role in the treatment of various cardiac disorders because of its actions on cardioprotection from hypoxia and oxidative stress. Studies on the role of carnitine administration to patients with myocardial infarction (MI), angina, and congestive heart failure generally have been positive. In general, treatment with carnitine (1.5 to 6 g/d for up to 1 year) results in a beneficial effect of fewer deaths and less heart failure when administered to patients after MI. Compared with placebo, carnitine use resulted in smaller increases in left ventricular end-systolic and end-diastolic volumes over time. In shorter term studies (1 to 3 months), carnitine therapy may have positive effects on symptoms of heart failure and angina in the post-MI period. Carnitine also seems to improve exercise tolerance and oxygen consumption in moderate to severe heart failure. Only preliminary results are available; results of a long-term (3-year) study should be reported soon. Studies specific to the dialysis population have generally shown that carnitine may have a beneficial effect on a number of cardiac parameters. Because cardiac disease is the most common form of death in patients with end-stage renal disease, these findings may be particularly important for this population. Moreover, because the relationship between conventional cardiac risk factors and cardiac disease is less clear in this population, the role of therapies that address pathological states specific to the dialysis population is worthy of study. Because a dialysis-related carnitine disorder is common among these patients, L-carnitine supplementation would be among these specific therapies. PMID- 12751053 TI - A review of the impact of L-carnitine therapy on patient functionality in maintenance hemodialysis. AB - Goals of maintenance hemodialysis therapy include not only the preservation of an individual patient's life in the presence of kidney failure, but also restoration of optimal quality of life. Although many conceptual and method problems are associated with the definition and assessment of quality of life in the chronically ill, there is broad agreement that patient quality of life is related to physical function and well-being. Evidence exists that despite advances in dialysis therapy, a high percentage of patients on maintenance dialysis therapy report chronic psychological symptoms, impaired activities of daily living and social function, and incomplete occupational rehabilitation that impair their functionality. This article reviews the impact of L-carnitine therapy on several dimensions of functionality in maintenance hemodialysis patients. PMID- 12751126 TI - Negative, neutral, and discordant results: what should an editor do? PMID- 12751127 TI - Influence of race in heart failure and cardiac transplantation: mortality differences are eliminated by specialized, comprehensive care. AB - BACKGROUND: Differences in mortality are thought to exist between African Americans and Caucasians with heart failure. These differences may be due to a variety of factors, including differences in disease process, socioeconomic status, and access to health care. Additionally, little data exist on racial differences between these two groups after cardiac transplantation. This study examines a single center, urban experience in treating African Americans and Caucasians with heart failure and after cardiac transplantation. We hypothesize that treatment in a specialized, comprehensive heart failure/cardiac transplantation program results in similar survival between African Americans and Caucasians. METHODS: We retrospectively reviewed the Rush Heart Failure and Cardiac Transplant Database from July 1994 to August 2000. Variables analyzed in the cardiomyopathy patients included survival (until death, placement of left ventricular assist device or cardiac transplantation), number of hospitalizations per year, length of stay per year, and utilization of outpatient resources. Follow-up period was from initial visit to death, transplantation, or implantation of left ventricular assist device. In those who underwent cardiac transplantation, we examined rejection rates (cellular and humoral), rejection burden, hospitalization data, and 5-year survival. A subgroup bridged to cardiac transplantation with a left ventricular device was also analyzed. RESULTS: Seven hundred thirty-four cardiomyopathy patients were identified: 203 were African Americans and 531 were Caucasians. The etiology of cardiomyopathy was more commonly ischemic in Caucasians as compared to non-ischemic in African Americans (P <.01). African Americans had more admissions to the hospital per year compared with Caucasians, 1.2 +/- 2.1 versus.5 +/- 1.1 (P <.01) with longer length of stay per year, 1.4 +/- 25.2 days versus 4.4 +/- 14.3 days (P <.01). Utilization of outpatient resources was significantly higher in African Americans compared with Caucasians with more use of continuous inotropes (13% versus 6%, P <.01), intermittent inotropes (11% versus 5%, P <.01), and home nursing after hospital discharge (52% versus 32% of hospital discharges, P <.01). Survival by Kaplan Meier analysis was comparable between the two groups (mean survival 1,470 +/- 72 days in African Americans versus 1521 +/- 46 days in Caucasians, log rank test [P =.6]). During this time, 30 African Americans and 73 Caucasians underwent cardiac transplantation. Fifty-three were bridged to transplantation with a left ventricular assist device (20 African Americans, 33 Caucasians). There were no differences in 5-year survival by Kaplan-Meier analysis despite higher peak preoperative panel reactive antibody levels in African Americans versus Caucasians (12% +/- 30% compared with 5% +/- 15%, P =.04), more overall treated rejection episodes per year in the African Americans (P <.01), as well as more posttransplant hospitalizations (2.2 +/- 1.2 times per year as compared with 1.7 +/- 2.1 times per year, P =.04). CONCLUSION: Delivery of care to heart failure patients in a comprehensive, specialized program results in similar survival regardless of race despite higher utilization of inpatient and outpatient resources. The finding that, after cardiac transplantation, African Americans do not have higher mortality rates, despite having higher rates of rejection overall and more hospitalizations, further supports the hypothesis that optimal care can improve outcomes despite unfavorable baseline clinical characteristics. PMID- 12751128 TI - Isolated bundle branch block and left ventricular dysfunction. AB - BACKGROUND: Population studies have identified the presence of bundle branch block (BBB) as a risk factor for increased mortality in patients with known cardiac disease. It is unknown how chronic ventricular dyssynchrony resulting solely from conduction delay affects ventricular function. METHODS AND RESULTS: Subjects were retrospectively identified from the University of California, San Francisco, electrocardiogram database with QRS prolongation. Subjects who had no evidence of heart disease and had baseline and serial studies evaluating ongoing risk of coronary artery disease status and left ventricular function were followed over time. Documentation of a minimum of 2 serial evaluations of cardiac status were present in a total of 51/176 (20%) subjects with isolated BBB who constitute the study population. After a mean follow-up of 52 +/- 45 months adjusting for differences due to age, sex, the presence of hypertension, hyperlipidemia, and diabetes mellitus-left ventricular ejection fraction diminished by -7.3 +/- 12% per year in the isolated left BBB cohort versus -1.9 +/- 4% in isolated right BBB cohort and -1.1 +/- 3% reduction in the isolated nonspecific intraventricular conduction delay cohort (P =.019). CONCLUSIONS: Left bundle branch block itself may be a cause of ventricular dysfunction. PMID- 12751129 TI - Cardiac size, autonomic function, and 5-year follow-up of chronic heart failure patients with severe prolongation of ventricular activation. AB - BACKGROUND: Chronic heart failure is characterized by left ventricular dilation and abnormalities of cardiac autonomic function. Up to 20% of patients with chronic heart failure have QRS prolongation, which can lead to asynchronous left ventricular contraction. We tested the hypotheses that in patients with chronic heart failure, QRS > 150 ms is a risk factor for additional abnormalities of ventricular morphology, heart rate variability, and increased mortality. METHODS AND RESULTS: In 184 patients with left ventricular ejection fraction < 35%, QRS duration was > 150 ms in 53, and 150 ms, left ventricular end-diastolic and end-systolic diameters were greater than patients with QRS duration 150 ms had less low frequency R-R interval spectral power (P <.04). At 5 years 60% of patients with QRS > 150 ms had died compared with 35% of patients with QRS 150 ms have exaggerated disturbance of cardiac autonomic function, and left ventricular remodeling and significantly higher mortality than patients with QRS duration 40%) by age and sex. RESULTS: A total of 2,239 patients were included in this study. EF was known or measured in 59.2% of all admissions. EF was measured less often in women and with increasing age, but using logistic regression only increasing age (P <.0001) and not sex (P =.247) was associated with a lower frequency of EF measurement. Women were more likely than men to have preserved systolic function in all age groups older than age 65 (P <.001), but the frequency of preserved systolic function increased with age in both men and women. CONCLUSIONS: In Medicare patients discharged with a principal diagnosis of heart failure, only 59% have a measurement of EF. Women have EF measured less often than men; this is solely a function of the older age of women. Preserved systolic function increases in both men and women with CHF with increasing age, but women are far more likely than men to have preserved systolic function at all ages. PMID- 12751131 TI - Prognosis of congestive heart failure in patients with normal versus reduced ejection fractions: results from a cohort of 2,258 hospitalized patients. AB - BACKGROUND: Patients with congestive heart failure have an annual mortality of 10% to 20% depending on disease severity. Though one third of these patients have normal left ventricular (LV) ejection fraction (EF), their natural history is poorly defined. Small population-based studies have suggested a more benign prognosis for patients with preserved LVEF. However, prognosis in hospitalized patients, who form a higher risk group, is not known. METHODS: We investigated the survival patterns of 2,258 patients with a primary hospital discharge diagnosis of congestive heart failure between 1990 and 1999. Survival was analyzed and patients with normal and reduced LVEF were compared. RESULTS: Their age was 71 +/- 11 years, and 97% were men. There were 1,535 deaths over a mean follow up of 786 days. Of these, 963 (43%) patients had a normal LVEF (>/=55%). Patients with normal LVEF were of the same age as those with reduced LVEF, but had a lower prevalence of atrial fibrillation (20 versus 26%, P =.03), left bundle branch block (2 versus 12%, P <.0001), significant mitral regurgitation (5 versus 31%, P <.0001) and electrocardiographic evidence of myocardial infarction (38 versus 60%, P <.0001). Despite lesser comorbidities, they had a higher mortality hazard, with a 5-year survival of 22% compared with 28% for those with systolic heart failure (P =.007). Proportional hazards model showed presence of normal EF as a categoric variable to be an independent predictor of mortality in those with heart failure after correcting for age and rhythm. CONCLUSIONS: Prognosis of hospitalized patients with congestive heart failure and normal LVEF is worse than those with reduced EF despite lesser comorbidities. Studies addressing optimal management of these patients are warranted. PMID- 12751132 TI - Prognostic implications of an initial loss of cardiac metaiodobenzylguanidine uptake and diabetes mellitus in patients with left ventricular dysfunction. AB - BACKGROUND: Although altered cardiac sympathetic innervation is related to fatal outcome, the mechanisms and prognostic value of an initial cardiac metaiodobenzylguanidine (MIBG) defect are not known. METHODS AND RESULTS: After quantitative cardiac MIBG imaging, 205 patients with left ventricular ejection fraction <50% were prospectively followed up with a primary end-point of cardiac death for 35 months. In regard to 38 cardiac deaths, consisting of 25 pump failure deaths, 11 sudden deaths, and 2 fatal acute myocardial infarctions, multivariate analysis identified diabetes mellitus as a significant independent predictor as well as reduced cardiac MIBG activity, use of nitrate, and New York Heart Association functional status. Independent of washout kinetics and cardiac function, patients with profound loss of initial MIBG uptake and those with late phase MIBG activity of 1.74 or less had significantly greater mortality rates than did their counterparts. Initial cardiac MIBG activity closely correlated inversely with annual cardiac death rate. CONCLUSIONS: An initial cardiac MIBG defect and presence of diabetes mellitus indicate a low probability of long-term survival. The profound loss of initial MIBG activity is likely to be due to structural deficit of sympathetic neurons themselves, rather than accelerated sympathetic function, suggesting that denervation is one of mechanisms of cardiac sympathetic dysfunction leading to lethal clinical outcomes. PMID- 12751133 TI - Regression of left ventricular hypertrophy after treatment of hypertension: comparison of directed M-echocardiography with magnetic resonance imaging in quantification of serial mass changes. AB - BACKGROUND: This study sought to validate the reliability of serial echocardiographic measurements in detecting left ventricular (LV) hypertrophy regression by using magnetic resonance imaging (MRI) as a reference standard. METHODS AND RESULTS: We studied a small population (n = 20) of patients enrolled in the Prospective Randomized Enalapril Study Evaluating Regression of Ventricular Enlargement (PRESERVE) trial for evaluating LV hypertrophy regression. LV mass was measured by both echocardiography and MRI at baseline and after 1 year. As compared with baseline, systolic and diastolic blood pressures were significantly decreased after 1 year (all P <.05). Echocardiographic technique showed an overestimation of LV mass by 27.6 g at baseline (P =.005) and by 37.1 g after 1 year (P <.001), and there were wide 95% limits of agreement (+/ 36.0 g at baseline; and +/-27.6 g after 1 year) when compared with MRI measurement. Significant changes of LV mass from baseline of -20 +/- 22 g (P<.01) and -29 +/- 19 g (P <.01) were detected by using echocardiography and MRI after 1 year, respectively (P =.02), and there were similarly wide limits of agreement for change in LV mass (+/-24.2 g). CONCLUSIONS: Despite the use of careful methodology, echocardiographic measurement of LV mass at a single time point or for serial studies resulted in significant variation in LV mass estimates from measurement using MRI. PMID- 12751134 TI - Left ventricular response to sustained volume overload from chronic aortic valve regurgitation in rats. AB - OBJECTIVES: Aortic regurgitation (AR) induces left ventricular (LV) eccentric hypertrophy in response to chronic volume overload. Patients suffering from this disease often remain asymptomatic for decades before progressive LV dysfunction develops silently. Because of this slow evolution, large clinical trials with long-term follow-up on subjects with chronic AR are hard to perform. To overcome this problem, animal models have been developed in the past but results were very heterogeneous. METHODS: Helped by echocardiography, we refined a known technique to induce homogeneous degrees of severe AR in Wistar-Kyoto rats. The effects on LV function without treatment and with nifedipine (25 mg/kg daily) (a drug currently recommended in humans with chronic AR) were evaluated by echocardiography. RESULTS: Over 6 months, nontreated animals developed progressive LV dilatation and eccentric hypertrophy, characteristic of chronic LV volume overload. The animals also developed progressive LV systolic dysfunction, mimicking closely the evolution of the disease in humans. Abnormal filling parameters were also detected in the majority of animals. Systolic and diastolic abnormalities were prevented but only partially in the group treated with nifedipine. CONCLUSION: This model can be used to study chronic AR and LV dysfunction associated with the disease. Nifedipine seems to protect the LV against chronic volume overload but only partially. Treatment strategies currently used in humans deserve further investigation. PMID- 12751136 TI - A broader paradigm for understanding and treating heart failure. AB - Several recent clinical trials in heart failure have failed to demonstrate improvements in survival, suggesting that new approaches must be adopted if there are to be further improvements in heart failure treatment. We propose a broader paradigm for heart failure care based on a chronic rather than an acute illness model. This approach recognizes a broader range of outcomes including patient perceived health status, expanded populations of interest that more closely reflect the changing epidemiology of heart failure, and a wider scope of interventions that address co-morbidity and health behavior. This approach will require a rationally designed health care delivery system that applies the best available evidence to the care of individual patients with heart failure. PMID- 12751135 TI - Potentiation of atrial contractility by paired pacing augments ventricular preload and systolic performance. AB - BACKGROUND: Paired electrical stimulation and postextrasystolic potentiation (PESP) of contractility has been extensively studied in ventricular myocardium, but less is known about PESP of atrial contractility. Our aim was to determine whether PESP of atrial contractility could augment left ventricular (LV) preload and improve LV systolic performance. METHODS AND RESULTS: A paired electrical stimulus closely following the pacing stimulus was applied to isolated atrial and ventricular myocardium from 4 dog hearts, and the interval dependent force potentiation was examined. In isolated atrial myocardium, paired pacing increased the active tension from a baseline of 1.36 +/- 0.23 to 2.60 +/- 0.57 g/mm(2); in ventricular myocardium active tension increased from 2.58 +/- 0.42 to 3.81 +/- 0.27 g/mm(2) (both P <.01). Then, LV pressure (micromanometer) and segment length (ultrasonic crystals) were measured in the intact hearts of 7 anesthetized dogs in which premature stimuli were applied to the atrium. In intact hearts, paired pacing of the atrium (coupling interval 200 ms) increased LV end-diastolic pressure from 3.8 +/- 1.0 to 6.4 +/- 1.0 mm Hg; systolic pressure increased from 105 +/- 6 to 112 +/- 7 mm Hg (both P <.05). LV pressure-length loop area (regional stroke work) increased 10.5 +/- 0.2%. CONCLUSIONS: Isolated atrial myocardium exhibits substantial PESP of contractility, which is similar to ventricular myocardium. In the intact heart, atrial PESP augments LV systolic performance by effecting an increase in LV preload. This technique may provide a means of improving cardiac performance in patients with heart failure. PMID- 12751137 TI - Treating chronic kidney disease. PMID- 12751138 TI - Managing chronic kidney disease. PMID- 12751139 TI - Preparing MCOs to manage chronic kidney disease. AB - Managed care organizations have begun recognizing the critical need to start implementing changes that will significantly improve diagnosis and treatment of chronic kidney disease (CKD). Alan Wright's perspective on managed care's shifting focus relative to managing these patients, presented here, ignited a lively discussion among panel members in the roundtable discussion that follows this section. By drawing on the strength of the data and information shared by Drs. Amedia and Provenzano, this article highlights how managed care can address the need for early identification of patients at heightened risk for CKD and points out ways to avoid the hidden costs associated with the various conditions that are related to renal disease. In addition, this article delineates ways that MCOs can strengthen educational efforts aimed at primary care physicians about early diagnosis and management of these patients. PMID- 12751141 TI - COPD gaining favor as focus for DM initiatives. PMID- 12751142 TI - Physician group builds tools to help practitioners boost diabetes outcomes. AB - A group practice has developed two new tools to help physicians and patients take DM matters into their own hands. The Diabetes Patient Report Card is designed to help patients stay on track with periodic blood testing and other routine preventive tasks, and the Diabetes Physician Audit Tool provides a simple way for physicians to measure their own performance on a range of measures. PMID- 12751143 TI - Incentives boost use of care management processes for treatment of chronic disease. AB - New research suggests that few physician organizations are investing in care management processes that are critical to improving quality and reducing costs for the chronically ill. Find out what incentives can do to turn the situation around. PMID- 12751144 TI - Immunotherapy: new guidelines suggest a 'window' for prevention. AB - New guidelines suggest that immunotherapy can, in some cases, actually prevent the development of allergic asthma in children with allergic rhinitis. The new recommendations offer advice on when to consider immunotherapy, optimal dosing, and how to handle and store medicines. PMID- 12751145 TI - New research establishes importance of specialty care after discharge for an MI. AB - While numerous studies have focused on what type of care is optimal for MI patients while hospitalized, little focus has been paid to what happens after discharge. New research shows that patients who continue to see a cardiologist in the early months after discharge have better outcomes than those who do not. Further, this research raises some disturbing questions about why certain groups of patients are less likely to receive this specialty care than others. PMID- 12751147 TI - Cell phones and EMI. Frequently asked questions. PMID- 12751146 TI - Replacing mercury-column sphygmomanometers. AB - Mercury-based devices have a long history of use in the healthcare setting. However, mercury poses a significant occupational and environmental exposure hazard. In recognition of this hazard, the American Hospital Association and the U.S. Environmental Protection Agency have set a goal to eliminate mercury in healthcare by 2005. As a result, healthcare facilities are now under increasing pressure to replace mercury-based equipment. Thermometers and sphygmomanometer columns are the most prevalent sources of metallic mercury in the healthcare setting. Many hospitals have already swapped mercury thermometers for electronic devices. Replacing mercury sphygmomanometers, however, has been more problematic, as concerns have been raised about the accuracy of the alternatives. Despite these concerns, some hospitals have been able to eliminate or significantly reduce the presence of mercury sphygmomanometers. In this Guidance Article, we describe factors that healthcare facilities should consider when choosing from among the available blood pressure measurement methods, and we provide practical advice for facilities trying to make the switch from mercury sphygmomanometers to alternative methods. PMID- 12751148 TI - Clarifying JCAHO's alarm-safety goal. A conversation with Richard J. Croteau, MD. PMID- 12751149 TI - Health technology slide shows. Web feature gets down to basics. PMID- 12751150 TI - Hazard report. Close-range EMI sends Nellcor Puritan Bennett 840 ventilators into "vent inop" mode. PMID- 12751151 TI - Hazard report. ECG leads-off alarms shouldn't be a low priority. PMID- 12751152 TI - User experience network. Rayovac batteries in Baxter Colleague pumps may overheat and swell. PMID- 12751153 TI - User experience network. Casters may loosen on Olympus TI-A1 endoscopy carts. PMID- 12751154 TI - Talk to the specialist. Negatively pressurized operating rooms. PMID- 12751155 TI - Best-case series for the use of immuno-augmentation therapy and naltrexone for the treatment of cancer. PMID- 12751156 TI - [Thirty years since the smallpox epidemic in Yugoslavia]. PMID- 12751157 TI - [Relation between body fat mass and lipid status in the obese working population]. AB - INTRODUCTION: Obesity can be defined as an excessive accumulation of health threatening body fat, caused by positive energetic balance. It can be classified according to body mass index as normal body mass, excessive body mass, significant obesity and extreme obesity. According to WHR (waist-hip ratio), it can be classified to android and gynoid type depending on fat tissue distribution. Android type has greater frequency of cardiovascular and metabolic complications, as is occurrence of premature atherosclerosis. As metabolic complications we consider lipid status disorders in obese workers, and these complications are related to body composition. MATERIAL AND METHODS: Among 331 workers, we separated 95 persons with BMI > 30 kg/m2. This group was classified according to gender, their body composition has been measured using bioelectrical impedance method and, subsequently, cholesterol, triglycerides, HDL and LDL levels were determined, searching for relationship of body composition with lipid status fractions p < 0.05. RESULTS: High body fatty was found in 33.03% of male and in 37.48% of female subjects. It was found that in male subjects cholesterol levels (6.70 mml/L), triglycerides (2.56 mml/L), limit values of LDL (3.93 mml/L) and limit values of HDL (1.16 mml/L) were highly risky. Positive insignificant relationship of body fatty mass with cholesterol, triglycerides, LDL, but not with HDL was found. DISCUSSION: Results point to highly risky limit values of lipid parameters in male and female subjects. These values can be explained by older age of subjects, their way of life and nutrition regimen, significant comorbidity in this group, and influence on working ability. CONCLUSION: It was found that obese male and female subjects have high values of body fatty mass. Male subjects have highly risky levels of lipid status fractions, while in female subjects these are limit values. Insignificant positive corelation of body fatty mass with lipid status fractions in tested population was found. The obtained results are significant regarding morbidity expectation and studies of complications of obese working population, as well as in judgment of working ability. PMID- 12751158 TI - [Acute oliguric renal failure in hypoxic neonates born at full term]. AB - INTRODUCTION: Acute renal failure (ARF) is a frequent clinical condition in neonatal intensive care units (NICU). The leading cause of neonatal ARF is perinatal asphyxia (PS). The aim of this study was to examine the relationship between the degree of PS and the severity of ARF in term neonates. METHODS: A prospective survey of 31 term neonates with Ps and but without congenital malformations or sepsis was performed in NICU of the regional Hospital of Gynaecology and Obstetrics in Belgrade (average number of deliveries about 6000 per year). ARF was diagnosed in the first 7 days of life when plasma creatinine was above 133 mumol/L for at least 48 hours while maternal renal function was normal. The degree of PS was determined according to Apgar score (AS) at 1 min. The severe PS was defined as AS < 3 and moderate PS as AS 4-6. RESULTS: Twenty neonates (64%) had oliguric ARF with urine output of 0.37 +/- 0.16 ml/kg/h while the others had nonoliguric ARF with urine output of 2.4 +/- 0.7 ml/kg/h. Most of neonates with oliguric ARF (65%) had severe perinatal asphyxia while in those with nonoliguric ARF moderate perinatal asphyxia predominated (73%). DISCUSSION: During hypoxic-ischaemic events many organs are injured, and the most vulnerable ones are kidneys and central nervous system. Our results showed a strong connection between perinatal asphyxia and A, which was in accordance with the results of other studies. Neonates with severe perinatal asphyxia had serious impairment of renal function, which was confirmed with strong correlation between Apgar score and plasma creatinine. In neonates with oliguric ARF, but not in those with nonliguric ARF, the highly positive linear correlations were found between AS and urinary output (r = 0.77; p < 0.01), plasma creatinine (r = 0.78; p < 0.01), fractional excretion of sodium (r = 0.76; p < 0.01), and index of renal failure (r = 0.80; p < 0.01). Only in oliguric neonates with severe perinatal asphyxia (31%) the outcome was fatal. CONCLUSION: We conclude that in tgerm neonates with severe perinatal asphyxia oliguric ARF was the predominant type of ARF. There is a good prediction of the severity of oliguric ARF according to the degree of perinatal asphyxia determined by Apgar score at 1 min. PMID- 12751159 TI - [Effect of postmenopausal hormone substitution on the lipid profile and coagulation factors in female smokers]. AB - Postmenopause and smoking impair lipid profile, induce hypercoagulability and reduce fibrinolytic capacity [1, 2]. Postmenopause induced lipid changes can be reversed by oestrogen replacement [3]. Oestrogens also reduce fibrinogen level [4] and have beneficial effects on endothelium [5]. Although several studies showed that hormone replacement therapy may increase the risk of thromboembolic diseases, procoagulant oestrogen activity has not clearly been demonstrated. It is well known that smoking accelerates oestrogen metabolism [6, 7], which may attenuate its beneficial effects. The present study was undertaken to determine if there is difference in beneficial effects of oestrogens between smokers and non-smokers in terms of coagulation process and lipids. The examination was a longitudinal, one-year, before/after therapeutic study, which included healthy postmenopausal women (FSH levels at least 40 U/l), 30 smokers and 32 non-smokers who were under 55 years of age and postmenopausal period shorter than 5 years. Women with surgically induced menopause received unopposed oral oestrogens, while those with spontaneous menopause were treated with combined oral oestrogen/progestogen therapy. Before entering the study and in three-months intervals total LDL, HDL cholesterol, triglycerides and VLDL were determined, as well as plasma fibrinogen, prothrombin time, and activated partial thromboplastin time. Neither beneficial nor adverse effects of oestrogens on lipids and coagulation were observed during one-year follow-up in smokers, although subjects with longer smoking history had higher triglycerides levels after 12 months of therapy. On the contrary, oestrogen replacement reduced total and LDL cholesterol, and increased HDL cholesterol in non-smokers, with no change in triglycerides and VLDL level. A decrease in fibrinogen levels and coagulation activity, expressed by prothrombin time and partial thromboplastin time, were also observed in hormone replacement therapy in postmenopausal women who did not smoke. Peroral hormone replacement therapy does not induce favorable lipid changes in smokers. Higher triglycerides levels observed after one-year therapy in women with a longer smoking history indicate that transdermal replacement maybe more suitable in this group. Peroral oestrogen replacement has no anticoagulant or procoagulant activity in smokers during the early postmenopausal period. PMID- 12751160 TI - [A rapid detection of motor block in patients with Parkinson disease during volitional movements of the hand]. AB - INTRODUCTION: An algorithm to study hand movements in patients with Parkinson's disease (PD) who experience temporary, involuntary inability to move a hand have been developed. In literature, this rather engimatic phenomenon has been described in gait, speech, handwriting and tapping, and noted as motor blocks (MB) or freezing episodes. Freezing refers to transient periods in which the voluntary motor activity being attempted by an individual is paused. It is a sudden, unplanned state of immobility that appears to arise from deficits in initiating or simultaneously and sequentially executing movements, in correcting inappropriate movements or in planning movements. The clinical evaluation of motor blocks is difficult because of a variability both within and between individuals and relationship of blocks to time of drug ingestion. In literature the terms freezing, motor block or motor freezing are used in parallel. AIM: In clinical settings classical manifestations of Parkinson's Disease (akinesia, bradykinesia, rigidity, tremor, axial motor performance and postural instability) are typically evaluated. Recently, in literature, new computerized methods are suggested for their objective assessment. We propose monitoring of motor blocks during hand movements to be integrated. For this purpose we have developed a simple method that comprises PC computer, digitizing board and custom made software. Movement analysis is "off line", and the result is the data that describe the number, duration and onset of motor blocks. METHOD: Hand trajectories are assessed during simple volitional self paced point-to-point planar hand movement by cordless magnetic mouse on a digitizing board (Drawing board III, 305 x 457 mm, GTCO Cal Comp Inc), Fig. 1. Testing included 8 Parkinsonian patients and 8 normal healthy controls, age matched, with unknown neurologic motor or sensory disorders, Table 1. Three kinematic indicators of motor blocks: 1) duration (MBT%); 2) onset (t%); and 3) number (N) of MB episodes, allow identification and quantification of motor blocks. Duration of motor blocks (MBT) is defined as the time sequence when (x,y) coordinates do not change their values and is expressed in percentage from the whole movement duration MBT% = MBT/T (%). If during some movements more than one motor block occurs (N > 1) then this movement is decomposed. The whole movement motor block (mbt) is the sum of all motor blocks MBT; during the same movement and expressed in percentage from the whole movement duration mbt% = mbt/T (%). The onset of motor block (t) is determined with the beginning of motor block and expressed in percentage from the whole movement duration: t% = t/T (%). After the determination of kinematic indicators of motor blocks (MBT, N, t) for healthy controls, their mean values are calculated. Statistical package ANOVA is applied to determine statistical significance of the difference between PD patients and mean values from age matched control healthy group. PD patients are then classified into two groups: one group consisting of PD patients with motor blocks and the other without motor blocks, similar to healthy controls. RESULTS: Acquired movements are processed and analyzed. Fig. 2 is an example of hand trajectories. Time course of (x, y) coordinates indicates motor block appearance, Fig. 3. Detailed presentation of kinematic indicators of motor block (MBT, N, t) is in Fig. 4. Intra-subject variability of these parameters is presented in Figs 5, 6 and 7 for patient #3. The results for N show that 45% of all patients #3 movements had none motor blocks (N = 0); 20% had N = 1; 15% had N = 2; 11.5% had N = 3; 5.7% had N = 4; 0.3% had N = 5; 0.7% had N = 6; 0.3% had N = 7 and 1% had N = 8 motor blocks. The results for t% show that 3% of all patients' #3 blocks started at first quarter, 17% started in the second, 36% in the third, and 44% in the last quarter of movement. The results for MBT% show that 14.5% of all movements had MBT% in the range 0-5%; 56% had MBT% 5-10%; 22% had MBT% 10-15%; 5.5% had MBT% 15-20% and 2% had MBT% 20-25%. No block lasted more than 25% from the whole movement duration. Table 2 is the summary of mean variability for kinematic indicators of motor block (N, mbt%, t%) and for the movement duration T during a 7 day-testing of patients #3. The analysis of calculated data for eight tested PD patients revealed a significant difference (p < 0.01) between healthy controls and three PD patients; data on five PD patients were not significantly different (ns). This method clustered 3 PD patients in the group that experience motor blocks, while the rest were in the group without their significant occurrence. DISCUSSION: This algorithm is an additional instrument in classical evaluation of PD patients during their clinical evaluation and treatment. It provides to clinician a rapid feedback on the changes of voluntary hand movements in everyday progress of illness. Furthermore, this method could be of assistance for developing strategies to overcome motor blocks in arm movements at their beginning, as well as for the feedback of the success of drug therapy. PMID- 12751161 TI - [Preoperative radiologic evaluation of tumors of the renal parenchyma]. AB - Renal cell carcinoma is frequently a matter of urological interest. In recent years there were significant improvements regarding the earlier diagnosis, more precise preoperative staging and appropriate therapy. One hundred patients (42-78 years old) with the preoperative diagnosis of renal cell carcinoma were analyzed. Preoperative radiological evaluation included: transabdominal ultrasound, intravenous urography, computer-assisted tomography, and angiography. In all patients after radical nephrectomy pathohistological diagnosis was established and patients with the confirmed renal cell carcinoma tumour staging was performed. All histological findings were compared with the preoperative results of radiological examinations. Reliability of all of them is separately determined. Our results confirmed that the most efficient method of preoperative staging was computer-assisted tomography (accuracy 93%). Diagnostic methods that were previously used like intravenous urography and angiography, were not useful for routine diagnostic purposes. Ultrasound is a precise but not an enough informative diagnostic tool (accuracy 87%). Combine used of both ultrasound and contrast computer-assisted tomography is cost-effective, and an enough precise combination for everyday use. PMID- 12751162 TI - [Results of histopathologic findings of endometrial changes in metrorrhagia]. AB - Endometrium is a tissue which is in permanent changes in the active gynaecological period of a female. This can be followed-up through histological, custological and histochemical changes. In determining the morphologic changes and disturbances in endometrium, pathohistological examinations were and still are necessary and irreplaceable in defining the real causes of this phenomenon. The aim of our work is to show the pathohistological results of the material obtained by explorative curettage in metrorrhagia, to determine the aetiology of the pathological process in endometrium, which provokes irregular bleeding, and to check the possible increase in precancerogenic and malignant endometrial conditions over a period of three years. The examinations covered 786 patients who asserted that bleeding was the main symptom of their illness. In all of them explorative curettage was performed mainly in diagnostic and often in therapeutic purposes. The number of examinations are in constant increase. The age of the largest number of patients was from 41 to 50 years, a period when the anovulary cycle is most frequent which, in turn, causes changes in endometrium which results in irregular bleeding i.e. in metrorrhagia (Tables 1, 2 and 3). The most frequent histopathological changes were manifested by adenomatic hyperplasia (160), then glandular (136) and cystic hyperplasia (92). Eleven cases of endometrial carcinoma were detected. In two of these patients the finding was not confirmed by pathohistological result after the operation or repeated explorative currettage. In 64 patients atypical hyperplasia i.e. Ca in situ was found. Acute endometritis followed by irregular bleeding was observed during inflammatory process of the internal genital organs. In 35 patients chronic endometritis was found after curettage following the extraction of the intrauterine device (in one case after 40 years of carrying). In 103 patients with hyperplasia myomatous changes were noted; this finding correlated with hyperoestrogenism, which is usually present in both disorders. All authors agree that pathohistological examination of endometrium is the most reliable in the search for a correct diagnosis. Despite of all modern diagnostic and technical means, difficulties always arise in the assessment of malignant potential of endometrial hyperplasia. Therefore, the patients with recurrent bleeding caused by hyperplasia should be under intensive gynaecological control. PMID- 12751164 TI - [Acute deafness--personal experience with therapy]. AB - The article deals with the results of treatment of 40 patients with acute deafness of unknown aetyology. Better results were obtained in patients with small degree of loss of hearing without vestibular symptomatology and in patients in whom therapy was started earlier. We suggest the early confirmation of diagnosis and urgent use of infusion therapy which improves the perfusion of the inner ear. PMID- 12751163 TI - [Brain abscesses of otogenic origin]. AB - Chronic inflammation of the middle ear is the most frequent cause of otogenic complications. Meningitis is the most frequent otogenic intracranial complication, followed by otogenic brain abscesses, while other complications are significantly less frequent. The study is aimed at presenting clinical causitry of otogenic brain abscesses consequential to chronic suppurative otitis in order to evaluate modern diagnostic and therapeutic possibilities. The study was retrospective and included the patients treated at the Institute of Otorhinolaryngology and Maxillofacial Surgery of the Clinical Centre of Serbia diagnosed with otogenic brain abscess during a five-year period (1996-2000). A total of 9 patients (male to female ratio 8:1), aged 16-68 years, were assessed. The following parameters were analyzed: sex, age groups, place of living, occupation, number of hospitalizations, diagnostic procedures, symptoms and clinical signs of otogenic complications, other otogenic complications associated with brain abscess, endocranial localization of otogenic abscess, therapeutic procedures (oto-surgical treatment) and intraoperative otological findings. In our group of patients, otogenic brain abscesses were significantly more frequent in male patients in their forties, with median age of 33.5 years. As for the place of living, the patients from the provinces were more frequent, while with respect to their level of education, those with elementary or high school degrees were predominant. The inflammatory process most frequently spread into the endocranium through direct destruction of the bone walls of the middle ear. Diagnostic procedures included history, clinical otorhinolaryngological examination, audiological and vestibulological assessment, neurological ophthalmologic and radiographic examinations (CT, MRI). CT is the most reliable diagnostic tool enabling localization of the change, timing of surgical treatment and monitoring of surgical success. Presence of other otogenic complications associated with brain abscess was evidenced in six of our patients. Cerebral localization of abscess was more frequent (7). Four patients underwent previous oto-surgical treatment. The treatment included primary neurosurgical approach (radical extirpation or abscess drainage), followed by radical oto-surgical treatment after improvement of the patient's general condition. PMID- 12751165 TI - [Rhabdomyosarcomas in the naso-paranasal area]. AB - The nasoparanasal region rhabdomyosarcoma belongs to the group of the rare malignant diseases, however it nevertheless represents major diagnostic and therapeutic problem. The study is aimed at presenting clinical casuistry of the nasoparanasal region rhabdomyosarcoma in adults and children, for the purpose of assessment of modern diagnostic and therapeutic possibilities. The study was retrospective and included the patients with evidenced and pathohistologically verified rhabdomyosarcoma of the nasoparanasal region diagnosed and treated at the Institute of Otorhinolaryngology and Maxillofacial Surgery of the Clinical Centre of Serbia, during the period 1988-2001, and subsequently at the Institute of Oncology in Belgrade. The total of 9 patients aged 3-50 years (median 15) were assessed, with the female to male ratio being 4:5. Seven patients were previously hospitalized in other health institutions. The following parameters were analyzed: sex, age group, number of hospitalizations, diagnostic procedures, histological tumour types, mode of treatment and disease outcome. In 7 patients, the tumour spread beyond the nasoparanasal region--in 6 into the orbit, in 5 into the epipharynx, in 3 intracranially, in 1 into the pterygopalatine pit and in another 1 into the parapharyngeal space. Regional lymph nodes were affected only in one patient, while distant bone metastases were evidenced on another one. Embryonal type of the rhabdomyosarcoma was verified in 7 patients, while two had the alveolar type. In the affected children mean survival was significantly longer (76.3 months) in comparison to the one observed in adults (11.5 months). Therapeutic procedure included combined application of the chemotherapeutic protocols (adjusted with respect to the age group and stage of the disease) and radiotherapy. Radical surgical procedures were applied on in case of stage I tumours, and they may also be used in rest-relapse cases subsequent to the applied chemo-, i.e., radiotherapy. PMID- 12751166 TI - [Juvenile angiofibroma of the nasopharynx--clinical diagnosis and therapy]. AB - Juvenile nasopharyngeal angiofibroma is an infrequent epipharyngeal tumour necessitating particular diagnostic and therapeutic procedures in comparison to other benign epipharyngeal tumours due to its expansive growth tendency. Our retrospective study is aimed at presenting clinical casuistry of the tumour in order to evaluate modern diagnostic and therapeutic possibilities. The study included 13 male patients, aged 13-24 years, who were hospitalized, diagnostically assessed and surgically treated at the Institute of Otorhinolaryngology and Maxillofacial Surgery, Clinical Centre of Serbia over the period 1990-June 2001. The following parameters were analyzed: sex, age groups, preoperative symptoms of the disease, diagnostic methods, embolization, local tumour spreading, number and time of tumour relapses and surgical approach. PMID- 12751168 TI - [Pyometra as a result of placement of an intrauterine device for 40 years and chronic endometritis]. AB - Pyometra is the accumulation of pus in the uterine cavity, thus stretching its walls and thinning and enlarging the uterus. This change is characteristic of the senium period of life, when the uterus is atrophic with a stenotic cervical canal. It most frequently occurs as a result of secondary infection of the cancerogenic tissue of the uterus and additional stenosation and clogging up of cervical canal by a malignant process. Apart from carcinomas of the body and uterus cervix, pyometra less often can occur in other illnesses such as senile endometritis and senile colpitis. As pyometra most frequently occurs in uterine carcinomas, in detection of this state, we must think of malignancy and direct our examination to this direction. In this paper we present a patient in whom pyometra developed because she carried an intrauterine device for forty years that resulted in chronic endometritis. The main symptoms for which the patient was admitted to hospital were abdominal pain and intensive suppurative vaginal excretion a month after removal of intrauterine device. The diagnosis of pyometra was made by gynaecological and ultrasound examinations, and also on the basis of gynaecological and ultrasound examinations one month after chronic endometritis was confirmed. On the basis of this finding we suspected that chronic endometritis was caused by this state. By cytological, PAP and histopathological examinations of samples obtained by explorative curettage and biopsy of the uterine cervix, malignant changes were eliminated a possible cause. By laboratory analysis and bacteriological examination of the uterine cavity and vaginal excretion, inflammatory changes of the uterine mocous membrane were confirmed as a cause of the pyometra. This conditions was due to carrying the intrauterine device for more decades. The therapy consisted of dilatation of the cervical canal and evacuation of accumulated suppurative contents and irrigation of uterine cavity with 3% solution of hydrogen peroxide and Povidon. Also wide spectrum antibiotics and uterotonics were given. Such treatment contributed to a fast and effectious recovery. PMID- 12751167 TI - [Giant liver abscess due to nearly asymptomatic choledocholithiasis]. AB - Solitary pyogenic liver abscess is usually caused by a metastatic infection through the portal blood flow or through the hepatic arterial blood flow from extra-abdominal pyogenic foci. Besides, it may be the result of local inflammatory diseases, such as cholecystitis, hydatid cyst, haematomas particularly with retained foreign bodies, etc. Suppurative cholangitis usually causes multiple pyogenic liver abscesses. Solitary pyogenic abscess is rarely caused by cholangitis, but practically always by suppurative cholangitis. Giant pyogenic liver abscess due to asymptomatic or mild cholangitis is a rarity. We present on a 63 year old man who developed a giant solitary pyogenic liver abscess in whom no other possible cause could be found or anticipated except practically almost asymptomatic choledocholithiasis accompanied with mild elevation of bilirubin content, alkaline phosphatase and gamma-GT. The patient was successfully treated operatively. Over 1800 ml. of pus was aspirated from the abscess cavity. Operative cholangiography performed in spite of the absence of gall bladder stones undilated and noninflamed common bile duct stone showed a small nonobstructing distal common bile duct stone. The duct was not dilated, the bile was clear and there were no signs of cholangitis in the inside of the common bile duct. Cholecystectomy and abscess cavity drainage led to uneventful recovery. The patient has been symptom-free for more than 3.5 years. PMID- 12751171 TI - [Is extrauterine pregnancy a complication of chlamydial infection?]. PMID- 12751170 TI - [Role of glucagon-like peptide 1 (GLP-1) in regulation of insulin secretion]. PMID- 12751169 TI - [Problems in diagnosis and extraction of foreign bodies from the orbit associated with pain]. AB - The aim of this study was to present the troubles and significance of a proper diagnosis of a foreign body which caused a double perforation of the eyeball and was retained in the orbit. Another reason for this case report was the intensive pain associated with the existing foreign body in the orbit. A male, 54 years old, had a perforating wound of the eyeball caused by a metal foreign body, which stayed in the orbit close to the scleral wall. X-ray and echographic examinations of the orbit were not conclusive regarding the question whether this foreign body was situated within or outside the eyeball. Only CT imaging showed that foreign body produced a double perforation of the eyeball and was externally close to the sclera. Foreign body was extracted by transconjunctival anterior orbitotomy through the inferior fornix, using the electromagnetic probe. Since the first day of injury and up to the eighth day, the patient had intensive deep orbital pain, which was alleviated only partially by analgetics. It could not be explained by secondary glaucoma because IOP was normal or by an inflammatory process, or in any other way. Only the extraction of foreign body from the orbit led to the complete relief of pain. We believe that the pain was caused by compression of foreign body to some of scleral sensory nerves. X-ray and echographic examinations of the orbit are not always a reliable proof in the proper evaluation whether foreign body is within or outside the eyeball in the orbit. Precise diagnosis can be made only by CT imaging. The pain in the orbit may be caused by compression of foreign body to sensory nerves. PMID- 12751172 TI - [Hernia as a sports injury]. PMID- 12751173 TI - [Changes do not come from legislation]. PMID- 12751174 TI - [Health risks of nurses in the workplace]. PMID- 12751176 TI - [Open fractures--nursing approach]. PMID- 12751175 TI - [Spiritual care in nursing practice observed in detail]. PMID- 12751177 TI - [Does the nurse feel/has difficulty in caring for the oncology patient?]. PMID- 12751178 TI - [Blood pressure measurement]. PMID- 12751182 TI - [New perinatal support plan]. PMID- 12751180 TI - [Youth suicidal acts are a not a choice]. PMID- 12751184 TI - [Disability and self image, the paths to dignity]. PMID- 12751183 TI - [From body to self image...]. PMID- 12751185 TI - [The construction of self in the eyes of others]. PMID- 12751186 TI - [Eulogy to weakness]. PMID- 12751187 TI - [Self concept in burned children]. PMID- 12751188 TI - [In Austria, women balance motherhood and work]. PMID- 12751189 TI - [2/6 Phototherapy]. PMID- 12751190 TI - [I--emergencies 6/enterocolitis]. PMID- 12751191 TI - Interview with Mack L. Richard. PMID- 12751192 TI - Thermonuclear attack--a presentation by Brigadier General Crawford Sams, 1957. PMID- 12751193 TI - Investigation of elevated whole body radiation monitoring results in a research environment. AB - When a personnel radiation monitoring device (badge) report indicates that a significant, elevated exposure has been received by an individual, an immediate and thorough investigation must be initiated to determine if the exposure is real, identity the source of the exposure, and to take appropriate steps to eliminate or control further exposures. The key points to consider in such an investigation are provided. PMID- 12751194 TI - Radiation protection program for declared pregnant workers at Los Alamos National Laboratory. AB - This article presents an overview of Los Alamos National Laboratory's (LANL) Fetal Radiation Protection Program (FRP) that satisfies the requirements set forth in 10 CFR 835 and LANL's Radiation Protection Program. At LANL, the FRP is one of three components of the larger Reproductive Health Hazards Program, which also includes Occupational Medicine and Industrial Hygiene. Although pregnant employees usually enter the program through Occupational Medicine, coordination with all three groups is achieved. The most important part of the FRP Program is performance of the workplace evaluation at the pregnant worker's workplace. At the meeting between the health physicist and the pregnant employee, the following topics are reviewed: risks to the embryo/fetus of working around sources of ionizing radiation; LANL's requirements and 10 CFR 835 regulations; her dose history; basic methods of radiation protection; and a detailed discussion of the work assignments/locations that enables the health physicist to complete an evaluation of the level of radiological hazards. Interface with her supervisor and the Operational Health Physics health physicist in charge of her work areas is essential in acquiring additional information. All of these data, including the radiation dose history and recommendations for possible work modifications or reassignment, are summarized in the workplace evaluation memo, which becomes part of the pregnant employee's medical file. Using input from LANL's legal staff, the author developed a document titled "Guidance to the Supervisors Regarding Fetal Radiation Protection and Reproductive Health Hazards," which instructs supervisors regarding the requirements and regulations, contact names for workplace evaluations, and, very importantly, how to avoid discriminatory behavior against pregnant employees. PMID- 12751195 TI - A quality educational program can significantly improve radiation safety. AB - The University of Cincinnati has greatly improved the quality of its radiation safety program, in part, by promoting education. The improved educational program includes radiation safety awareness level training for all employees. Individuals who will handle radioactive material and/or operate radiation-generating equipment receive additional training. Handlers of radioactive material go through an initial retraining program prior to handling any radioactive material, are required to attend annual continuing education retraining, and are provided ongoing corrective action education. References are also readily available to individuals on a web page maintained by the Radiation Safety Office. The minimum training provided to individuals who operate radiation-generating equipment is basic radiation safety training, machine specific instruction, and orientation to use appropriate safety measures. PMID- 12751196 TI - Development of a laser entryway safety control system in the research university setting. AB - Industry consensus guidance and good health physics practice dictate laser entryway safety controls for laboratories housing Class 4 laser controlled areas. The design and installation of an effective laser entryway safety control system presents several technical, administrative, and funding challenges. These challenges prompted the Radiation Safety Office at this institution to try a "winwin" approach involving utilization of free student labor to design, construct, and install one such system. This paper outlines the development and installation of the resulting laser entryway safety control system. PMID- 12751197 TI - Decontaminating the nasal passages. AB - Nasal decontamination may be indicated when the anterior nasal passages are contaminated with highly radioactive material or radioactive material with either irritating or toxic properties. Nasal irrigation (wash, rinse, douche, lavage) is an established technique used for other conditions and can be applied in these cases. This paper discusses the rationale and use of nasal irrigation and how to perform the technique. PMID- 12751198 TI - Shielding design for a PET imaging suite: a case study. AB - The introduction of positron emission tomography into the clinical environment presents the medical health physicist with another challenge to his/her shielding acumen. On one hand, elaborate models can be employed, but most of these are beyond the resources possessed by most institutions. On the other hand, one could perform the analysis using simplifying assumptions (e.g., point source geometry, with or without build-up). This kind of approach would likely overestimate the shielding requirements. Such over-design is not ALARA. In fact, over-design could place such tight engineering or cost constraints on a project as to make it untenable. Recently, this designer was faced with the need to design a PET imaging suite with both engineering and time constraints. This paper describes an approach using resources readily available to medical health physicists. By using the dimensions of the bottle manikin (BOMAB) phantom as a guide, a human-form source was developed. Combined with a point-kernel shielding code, the exposure environment was readily modeled and shielding recommendations developed. In addition, to validate the model, results from preoperational instrument surveys and integrating dosimeters are discussed. PMID- 12751199 TI - Preliminary evaluation of real time false CAM alarm prediction through continuous radon monitoring. AB - The outputs of two real-time radon monitors were compared with the output from a continuous air monitor (CAM) to evaluate the potential of predicting false CAM alarms. From the results obtained, it appears that the radon monitor will be able to indicate when large rapid changes in radon levels are occurring before the effect on a CAM has taken place. PMID- 12751200 TI - Laser accidents: occurrence and response. AB - The goal of the laser safety officer and any laser safety program is to prevent a laser accident from occurring, in particular, one that causes an injury to a person's eyes. Most laser safety courses "in-house commercial" discuss laser accidents, causes, and types of injury. Many do not address appropriate actions to be taken if such an accident occurs. This article reviews a sampling of laser accidents that have occurred and provides a quick overview of the appropriate steps to take when a laser accident occurs. PMID- 12751201 TI - Comment on article by Hogue. PMID- 12751202 TI - [Construction of shuttle expression vector and expression of thymosin alpha 1 in Synechococcus sp. PCC7942]. AB - The shutter expression vector pPREUT was constructed from the plasmid pPRS-1 containing the endogenous small plasmid of Plectonema boryanum. The expression elements such as heat shock gene groESL promoter, foreign gene Ub-thymosin alpha 1, rbcS polyA terminator and Kanamycin resistance gene were all included. The shutter plasmid pPREUT was directly transferred into Synechococcus sp. PCC7942. The transformants were obtained through Kanamycin screening. Southern blotting analysis showed that the shutter plasmid have been transferred into Synechococcus sp. PCC7942. After induction by heat shock(42 degrees C) for 30 min, the foreign protein UB-T alpha 1 was expressed efficiently, which reached 7.5% of total amount protein. PMID- 12751203 TI - [Expression and biological activity of human neurotrophin-3 in olfactory ensheathing cells mediated by retroviral vector]. AB - We successfully constructed a retrovirus expression vector pRev-TRE-NT3. The supernatant with highest retroviral titers was obtained by transfection of two retrovirus vector, pRev-TRE-NT3 and pRev-Tet-On, into an ecotropic Ecopack-293 cells. Primarily cultured rat olfactory ensheathing cells (OECs) were co-infected with retrovirus following pRev-TRE-NT-3 and pRev-Tet-On system of produced retrovirus. By adding different concentrations of Doxycline, genetically modified OECs were induced to secrete human NT-3. The secreted NT-3 in OEC culture supernatant was detected with western-blot, and its biological activity was confirmed by the growth bioassay of dorsal root ganglion (DRG) cells. The results are as follows: (1) 0.78 kb hNT-3 cDNA was harvested and successfully cloned into pRev-TRE vector by PCR and restriction enzyme digestion methods. The constructed plasmid was identified with electrophoresis, and the direction of hNT-3 cDNA inserting pRev-TRE vector was integrated correctly; (2) NT-3-modified OEC culture supernatant contained 28 kd NT-3 which can bind NT-3 antibody in western-blotting assay, and NT-3 expression was concentration-dependent on Doxycline. NT-3 was not detected in control groups; and (3) Numerous DRG cells migrated from DRG tissue mass in NT-3-modified OECs experimental groups, and these cells had long and thin processes with strong dioptre. These processes formed complicated networks. As for unmodified OEC control groups, a few DRG cells migrated from the tissue mass, and the processes of these cells were rather short. In blank groups, no cells migrated and grew. And their processes in control groups are significantly shorter than NT-3-modified OECs experimental groups. Our results indicate that Tet-On-regulated NT-3 expression in OECs was efficient, offering novel material of in vivo transplantation for the repair of CNS injury. PMID- 12751204 TI - [Redistribution of cell contents between declining old and growing young leaves in spring onion]. AB - During storage of a spring onion (Allium fistulosa var. dacong), the plant is unable to assimilate external resource, nutrient requirement of the new bud is entirely drawn from the withering leaf where both food reserves and protoplasmic constituents (i.e. organic complexes containing the essential mineral elements) are to be mobilized. Evidences were given to show that the withdrawal process of cell contents from the old part for establishing the new growth should be considered as an alternative mode or a supplementary measure of normal phloem transport of leaf photosynthates. Removal of cell contents from the declining old part to the developing new growth is of frequent occurrence in higher plants. In the present case, it followed the process of programmed cell death (PCD). That was examined and shown by electron microscopy and biochemical analysis. There appeared shrinkage of cell volume, dissolution of cell constitution and degradation of DNA. The present investigation has also shown that the withdrawal of cell contents from the declining leaf sheath of spring onion can be wholly exhaustive. It may be carried out by active movement of the partially degraded protoplasmic constituents in forms of fragments and vesicles across the tissue, until hardly anything in the cellulose framework of the dry papery sheath, except a few crystals of Ca2+. Presence of high ATPase and APase activities along the transport path also supports this view. PMID- 12751205 TI - [Photoautotrophic growth and photosynthetic characteristics of carrot callus]. AB - Photoautotrophic carrot callus was induced by gradually lowering the sucrose concentration in culture medium. In the course of induction, the net photosynthetic rate on chlorophyll basis of callus increased gradually with the decreasing of sucrose concentration in culture medium, and the net photosynthetic rate of photoautotrophic callus could even be higher than that of leaf. In the course of changing into photoautotrophic callus, its chlorophyll content increased gradually, and the dark respiratory rate and the ratio of Chl a/Chl b decreased gradually. Parallel with these was the development of the microstructure of thylakoid of callus. PMID- 12751206 TI - [Isolation, culture and biological character studies on human fetal pancreatic nestin positive cells]. AB - Pancreatic nestin positive cells are multipotential stem cells which may play an important role in the development of pancreas. Here we isolated nestin positive cells from human fetal pancreases and then examined its biological characters in vitro. The results showed that: (1) Nestin positive cells expressed high level transcript of ABCG2/BCRP1 which was the molecular determination of stem cells, and they differed from pancreatic ductal epithelium cells both in morphology and in growth behavior. (2) Upon confluence, these cells self assemble to form islet like cell clusters (ICCs). (3) During differentiation, nestin positive cells in ICCs expressed markers of different cell lineages which meant that they were multipotential, and they could generate insulin-producing cells after inducement. PMID- 12751207 TI - [Real-time quantitative PCR assay of Epstein-Barr virus in whole blood cells of nasopharyngeal carcinoma patients]. AB - EBV is detected in more and more tumors, and is relative to carcinogenesis. We studied the copies of EBV DNA in whole blood of NPC patients and healthy controls by real-time quantitative PCR. In the 73 NPC patients and 83 controls, the positive rate of EBV in blood of NPC is 46.6%, while 13.3% in control. The mean copy number is 3.9 x 10(4) copys/microgram DNA in controls, which is much higher than NPC patients (which is 1.7 x 10(5) copies/microgram DNA). EBV infection is relative with NPC, while the lytic form of EBV maybe more important than its latent form. These results suggest that whole blood EBV DNA may be a valuable tool for molecular diagnosis of NPC. PMID- 12751208 TI - [Roles of protein kinase C in mouse oocyte maturation and fertilization in vitro]. AB - Protein kinase C is an important family of serine/threorine protein kinase. In this experiment the possible roles of PKC in meiotic maturation, fertilization and egg activation were studied in mouse oocytes. Two isoforms of PKC, PKC alpha and beta 1, were localized in mouse oocytes. PKC activator PMA inhibited the resumption of meiosis in oocytes at GV stage, whose effect could be overcomed by PKC inhibitor calphostin C, neither the phospholipase C inhibitor U73122 nor the PKC delta-specific inhibitor rottlerin. The result of Western bolt indicates that both PKC alpha and beta 1 express stably during oocyte development. The translocation of PKCs and cortical granule exocytosis were observed by confocal microscopy. The results show that PKC alpha translocate from the cytoplasm to the membrane after fertilization or drug-stimulation, accompanied by CG exocytosis. This fact suggested that PKC alpha might be involved in the regulation of cortical reaction in mouse oocytes. Roles of PLC gamma in mouse oocyte during fertilization and localization of PKCs in mouse ovary were also investigated in this experiment. The result suggested that there did not exist a feedback regulation of PLC gamma on PKC during fertilization. PMID- 12751209 TI - [The influence of pregnancy and hormone on the lymphatic stomata in guinea pig's ovary bursa]. AB - In order to study the influence of pregnancy and hormone on the lymphatic stomata of guinea pig's ovary bursa, the lymphatic stomata of ovary bursa were observed during pregnancy and by using exogenous hormone, including serum gonadotrophin (PMSG) combined with human chorionic gonadotropin (HCG) to promote ovulation and androgen to inhibit the ovary. Then the lymphatic stomata in the inner layer of ovary bursa and their absorption functions were observed with scanning electron microscope (SEM) and trypan blue as a tracer respectively. The lymphatic stomata were also counted by the Elescope computer image processing system. We found that the quantity of tracer absorbed by the lymphatic stomata of ovary bursa in pregnant group was more than that in ovulation-promoted group, and least in the androgen-treated group, which had statistical significance between each couples of them (P < 0.05). The opening area of the lymphatic stomata in pregnant, ovulation-promoted and androgen-treated groups were 189.9 +/- 48.7 micron 2/1000 micron 2, 104.4 +/- 31.2 micron 2/1000 micron 2 and 40.5 +/- 18.7 micron 2/1000 micron 2 respectively, which had remarkable statistical difference between each couples (P < 0.01). Under SEM observation, the secretory granules in ciliated columnar epithelium were less in pregnant group than the other two, while both the secretory granules and the cilium were most active in ovulation-promoted group. The results indicated that both the openings and absorption functions of the lymphatic stomata in guinea pig's ovary bursa could be affected by pregnancy and exogenous homone. There existed relationship between ovary function and the regulation of the lymphatic stomata in ovary bursa. PMID- 12751210 TI - [Some factors affecting in vitro development of porcine embryo reconstitution from somatic cells nuclear transfer]. AB - In this paper, a study on reconstitution of porcine oocytes by using nuclear transfer with cumulus cells(CC) and fibroblast cells(FC) was carried out. Reconstituted oocytes which were the fusion with CC and showed a cleavage rate of 56.7%, developed to morula(11.7%) and blastocysts(6.7%) phases which were higher than those derived from the fusion with FC(p < 0.05). The results of this study also involved the effects of oocyte collection method and maturational age of recipient oocytes during the in vitro development of nuclear-transfer embryos which were reconstructed with cultured cumulus cells. The cumulus cells synchronized in G0/G1 phases through serum-starvation culture, were transferred into enucleated oocytes which were collected by aspiration or dissection method and cultured for 33 or 44 h. Reconstituted embryos were activated with a combination of calcium ionophore A23187 or electric pulsation and 6-DMAP, and cultured for 6 days. As for the oocytes collection methods, activation treatment in the presence of cytochalasin B did not affect the developmental rate of embryos reconstituted with 44-h-mature recipients. However, the development rate of reconstituted embryos with 33-h-mature recipients was significantly higher(p < 0.05) by activation with the combination of electric pulsation and 6-DMAP. These results suggest that reconstituted porcine embryos derived from cultured cumulus cells can develop to the blastocyst stage and that the development of the former could be improved by reconstruction with young oocyte cytoplast after the activation with the combination of electric pulsation and 6-DMAP. PMID- 12751211 TI - [Isolation and expression pattern analysis of rice auxin-induced eIF3a coding gene]. AB - Eukaryotic initiation factors (eIFs) are key elements in protein synthesis. Among identified 13 initiation factors, eIF3, composed of 8 or more subunits, is the largest one and plays a central role in the initiation of translation. As the largest subunit, eIF3a mediates most functions of eIF3. Based on the experiments that rice eIF3a gene could be induced by auxin (through fluorescent differential display PCR), a cDNA coding for rice eIF3a, named OseIF3a1, was isolated. OseIF3a1 cDNA, with a length of 3459 bp (including 5' and 3'-UTR) and encoding a 986-aa polypeptide, shared sequence identity of 82.4% and 70.1% with that in maize and tobacco. Comparison of cDNA and genomic sequence revealed the presence of 13 exons and 12 introns in OseIF3a1 gene. Reverse transcript PCR (RT-PCR) analysis indicated the expression of OseIF3a1 in root, shoots, young spike, stem and leaf tissues, promoter-GUS fusion studies further confirmed its expression in root tip, leaf, glume and stigma. The induction of OseIF3a1 by auxin suggested that function of auxin on growth may involve the regulation on translation level. PMID- 12751212 TI - Influence of substrate salinity on structure characteristics of petiole vessel elements of Bruguiera gymnorrhiza. AB - The anatomy of the mature leaf's petioles of one-year-old Bruguiera gymnorrhiza seedlings cultured in greenhouse was studied in this paper. The results are as follows: (1) Most of the vessel elements of the petioles of B. gymnorrhiza are of scalariform. Helical vessels and their transition type only account for a small portion of the total vessels. However, there is a notable increase in the number of helical vessels and their transition type with increasing salinity. (2) The length of petioles, diameter and length of scalariform vessel elements, and bar number of a scalariform perforation plate all exhibit of parabolic relationship with substrate salinity. They maximized at salinity between 20 g/L and 30 g/L. (3) It is new phenomenon that there are two scalariform perforation plates, facing differently, at the polar of the vessel element of the petioles in the seedlings under 10 g/L treatment. (4) Under low salinity, the changes in the structure of vessel elements had the tendency to promote the transportation of water with increasing salinity. While under high salinity, it tended to raise the reliability of the water transportation. The adaptive significance of the morphological characteristics of vessel elements in the petiole was discussed. PMID- 12751213 TI - [The formation process of chromatoid body in spermatogenic cell of teleost, Spinibarbus caldwelli (Nichols)]. AB - The formation process of chromatoid body in spermatogenic cell of teleost, Spinibarbus caldwelli (Nichols) was observed with electron microscopy. The chromatoid body was formed in the primary spermatogonium. First, the precursor of chromatoid body adhered to the inner surface of the nuclear envelope. The nuclear envelope where the chromatoid body adhering was destroyed. At the inner side of the precursor of chromatoid body, new nuclear envelope formed. It separated the precursor of chromatoid body from nucleus. The newly formed nuclear envelope was concaved. Thus a hollow appears on the surface of the nucleus. The precursor of chromatoid body gathered in the hollow and condensed into chromatoid body. There were many big gaps on the newly formed nuclear envelope. A little precursor of chromatoid body which had not been separated from the nucleus before comed out of the nucleus through these gaps and into the hollow, where it integrated with the chromatoid body. The way of chromatoid body formation in spermatogenic cell of Spinibarbus caldwelli was different from the way by which nuclear material comes out of the nucleus. It was suggested that there was a different way by which nuclear material could go out of the nucleus. The chromatoid body combined with mitochondria soon after it was formed, and left the hollow. During the development of the spermatogonium, the chromatoid body separated from mitochondria. PMID- 12751214 TI - [Floral homeotic variants of in vitro seedlings in Sinningia speciosa Hiern]. AB - Six types of floral homeotic variants of in vitro seedlings were observed in doubleflower sinningia. Type I, red and green mosaic petals exist in the outermost whorl of petal-whorls, 2.38%. Type II, the outermost whorl of petal whorls exhibit green petals with thin yellow edge, 25.0%. Type III, green petals exist in the innermost side of normal red petal whorls, 1.78%. Type IV, multiple whorls of green petals exist in the inner side of normal sepals, no stamen and carpel, 1.67%. Type V, it exhibits duplicated whorls of sepals in the outermost, 7.14%. Type VI, it exists multiple whorls of green sepals, no petal, stamen and carpel, 0.12%. The total percentage of all types of floral homeotic variants is up to 38.1%. The distribution of nodal site of homeotic flowers were analyzed, and the results showed that the homeotic flower occurred mainly at the fourth and fifth nodes. PMID- 12751215 TI - [Efficiency of the human visual system in discrimination of dynamic patterns]. PMID- 12751216 TI - [Changes in evoked potentials depending on attention level in performance of visual tasks]. PMID- 12751217 TI - [Changes in cortical activity during altered state of consciousness: study of meditation by high resolution EEG]. PMID- 12751218 TI - [Effect of arid climate conditions on psychophysiological performance of gas industry operators]. PMID- 12751219 TI - [Effect of examination stress on cardiac activity in students of schools providing differentiated education]. PMID- 12751220 TI - [Thoracic and abdominal respiration during the repetition of fixed respiration volumes under chemoreceptor stimulation]. PMID- 12751221 TI - [Relationship between mineral metabolism and blood rheology in athletes]. PMID- 12751222 TI - [Effect of erythrocyte acetylcholinesterase in hormonal regulation of adaptation to physical exercise]. PMID- 12751223 TI - [Effect of cold weather training on the thyroid gland and parameters of lipid metabolism in long-term residents of the northeast of Russia]. PMID- 12751224 TI - [Thermoregulation and blood circulation in adults during short term exposure to extreme temperatures]. PMID- 12751225 TI - [Physiological value of physical exercise and mitochondrial volume of working muscles in people exposed to long term hypokinesia. Effect of local resistance exercise ]. PMID- 12751226 TI - [Anatomical cross-sectional areas and volumes of the lower extremity muscles]. PMID- 12751227 TI - [Hemodynamics in patients with lower extremity fractures of different etiology]. PMID- 12751228 TI - [Effect of additional support on the maintenance of vertical posture in people]. PMID- 12751229 TI - [Diameter of the main arteries in the lower extremities after the bicycle ergometer test]. PMID- 12751230 TI - [Generation of nitric oxide by peripheral blood leukocytes and platelets in healthy people and in patient with thermal trauma]. PMID- 12751231 TI - [Pharmacological ethnic metabolic safety. Communication I]. PMID- 12751232 TI - [Vasilii Vasil'evich Parin--the founder of modern concepts of physiology and space biology (on the 100th anniversary of his birthday)]. PMID- 12751233 TI - [Electrooculographic system for the recording and two-dimensional mapping of concomitant eye movement]. PMID- 12751234 TI - [Specific oxygen consumption in the myocardium of patients with congenital and acquired heart defects]. PMID- 12751235 TI - [Impairment of the body tolerance to its own microflora--type of immunopathology]. PMID- 12751236 TI - [The biology of aerobic methylobacteria capable of degrading halomethanes]. AB - Recent data on the biology of aerobic methylotrophic bacteria capable of utilizing toxic halogenated methane derivatives as sources of carbon and energy are reviewed, with particular emphasis on the taxonomic, physiological, and biochemical diversity of mono- and dihalomethane-degrading methylobacteria and the enzymatic and genetic aspects of their primary metabolism. The initial steps of chloromethane dehalogenation to formate and HCl through a methylated corrinoid and methyletrahydrofolate are catalyzed by inducible cobalamin methyl transferase, made up of two proteins (CmuA and CmuB) encoded by the cmuA and cmuB genes. At the same time, the primary dehalogenation of dichloromethane to formaldehyde and HCl is catalyzed by cytosolic glutathione transferase with S chloromethylglutathione as an intermediate. The latter enzyme is encoded by the structural dcmA gene and is under the negative control of the regulatory dcmR gene. In spite of considerable progress in the study of halomethane dehalogenation, some aspects concerning the structural and functional organization of this process and its regulation remain unknown, including the mechanisms of halomethane transport, the release of toxic dehalogenation products (S-chloromethylglutathione, CH2O, and HCl) from cells, and the maintenance of intracellular pH. Of particular interest is quantitative evaluation of the ecophysiological role of aerobic methylobacteria in the mineralization of halomethanes and protection of the biosphere from these toxic pollutants. PMID- 12751237 TI - [Physiology of organotrophic and lithotrophic growth of the thermophilic iron reducing bacteria Thermoterrabacterium ferrireducens and Thermoanaerobacter siderophilus]. AB - Growth physiology of the iron-reducing bacteria Thermoterrabacterium ferrireducens and Thermoanaerobacter siderophilus was investigated. The stimulation of the organotrophic growth of T. ferrireducens and T. siderophilus in the presence of Fe(III) was shown to be due to the utilization of ferric iron as an electron acceptor in catabolic processes and not to the effect exerted on the metabolism by Fe(II) or by changes in the redox potential. It was established that Fe(III) reduction in T. ferrireducens is not a detoxication strategy. In T. siderophilus, this process is carried out to relieve the inihibitory effect of hydrogen. T. ferrireducens was shown to be capable of lithoautotrophic growth with molecular hydrogen as electron donor and amorphous ferric oxide as electron acceptor, in the absence of any organic substances. The minimum threshold of H2 consumption was 3 x 10(-5) vol % of H2. The presence of CO dehydrogenase activity in T. ferrireducens suggests that CO2 fixation in this organism involves the anaerobic acetyl-CoA pathway. T. siderophilus failed to grow under lithoautotrophic conditions. The fact that T. ferrireducens contains c-type cytochromes and T. sidrophilus lacks them confirms the operation of different mechanisms of ferric iron reduction in these species. PMID- 12751238 TI - [Induction of cytochrome P-450 and ethanol oxidation in Yarrowia lipolytica]. AB - The study of the effect of different ethanol concentrations in the medium on the growth and the activity of enzymatic systems involved in ethanol oxidation in Yarrowia lipolytica showed that the cultivation of yeast cells on 1 and 2% ethanol caused their rapid growth and a drastic increase in cell respiration and sensitivity to cyanide already in the first hours of cultivation. At the same time, during cultivation on 3, 4, and 5% ethanol, the growth and respiration of yeast cells were considerably suppressed. All of the ethanol concentrations studied induced the synthesis of cytochrome P-450, its dynamics in cells being dependent on the initial concentration of ethanol in the medium. When the initial concentration of ethanol was 1 and 2%, the content of cytochrome P-450 in cells steeply decreased after a short period of induction. But when the initial concentration of ethanol in the medium was 3 to 5%, the content of cytochrome P 450 in cells was high throughout the cultivation period. The induction of cytochrome P-450 in cells preceded the induction of the NAD-dependent enzymes alcohol dehydrogenase and catalase, which, like cytochrome P-450, are also involved in ethanol oxidation by yeasts. The activity of catalase was higher in the yeast cells grown in the presence of 3 to 5% ethanol than in the cells grown in the presence of 1 and 2% ethanol. The roles played by cytochrome P-450, alcohol dehydrogenase, and catalase in ethanol oxidation by yeast cells are discussed. PMID- 12751240 TI - [The Penicillium commune Thom and Penicillium clavigerum Demelius fungi- fumigaclavines A and B producers]. AB - The type strains Penicillium clavigerum VKM F-447 and P. commune VKM F-3233 are found to produce fumigaclavines A and B. Of the seven other strains of these species, only two strains, P. commune VKM F-3088 and F-3491, possess the ability to synthesize these alkaloids. It is suggested that the five other strains under study either lost such an ability or require very specific conditions for the synthesis of these alkaloids. PMID- 12751239 TI - [Effect of cytochrome oxidase inhibitors on the yeast thermotolerance]. AB - The investigation of the effect of the cytochrome oxidase inhibitors sodium cyanide and sodium azide on the thermotolerance of the yeasts Rhodotorula rubra, Debaryomyces vanriji, and Saccharomyces cerevisiae showed that these inhibitors diminish the thermotolerance of R. rubra and D. vanriji, but do not affect the thermotolerance of S. cerevisiae. Taking into account the fact that, unlike the latter yeast, R. rubra and D. vanriji are nonfermentative yeasts, the difference in the effects of the inhibitors on the yeast thermotolerance can be readily explained by the different types of glucose utilization (either oxidative or fermentative) in these yeasts. The data obtained also provide evidence that there is a correlation between the functional activity of mitochondria and the thermotolerance of yeast cells. PMID- 12751241 TI - [Mechanisms of the alkaloid aurantioclavine excretion and uptake during Penicillium nalgiovense VKM F-229 growth]. AB - The biphasic dynamics of the alkaloid aurantioclavine in the culture liquid of Penicillium nalgiovense VKM F-229 is shown to be due to the diauxic growth of the fungus on two carbon sources, succinate and mannitol. In the phase of active growth on succinate, the fungus synthesizes aurantioclavine and excretes it into the medium in an energy-independent manner, as a result of which the concentration of the alkaloid in the culture liquid rises. During the phase of metabolic adaptation to the other carbon source, mannitol, the concentration of aurantioclavine in the culture liquid falls, probably due to the energy-dependent uptake of the alkaloid by fungal cells. The reversible excretion of aurantioclavine in P. nalgiovense indicates that these are regulated processes and depend on the growth parameters and the physiological state of the fungus. PMID- 12751242 TI - [Teichoic acids in the cell walls of Microbispora mesophila Ac-1953t and Thermobifida fusca Ac-1952t]. AB - The cell walls of Microbispora mesophila strain Ac-1953T (the family Streptosporangiaceae) and Thermobifida fusca Ac-1952T (the family Nocardiopsiceae) were found to contain teichoic acids of a poly(glycerol phosphate) nature. The teichoic acid of M. mesophila (formerly Thermomonospora mesophila) represents a poly(glycerol phosphate) containing 5% of substituent 2 acetamido-2-deoxy-alpha-galactosaminyl residues. The teichoic acid of such kind was found in actinomycetes for the first time. The cell wall of T. fusca (formerly Thermonospora fusca) contains two teichoic acids, namely, unsubstituted 1,3-poly(glycerol phosphate) and beta-glucosylated 1,3-poly(glycerol phosphate). PMID- 12751243 TI - [Fluorene cometabolism by Rhodococcus rhodochrous and Pseudomonas fluorescens]. AB - The transformation of fluorene by Rhodococcus rhodochrous strain 172 grown on sucrose and Pseudomonas fluorescens strain 26K grown on glycerol was studied as a function of the substrate concentration and the growth phase. Under certain cultivation conditions, fluorene was completely consumed from the medium. The specific transformation rate of fluorene was considerably higher when it was transformed in the presence of the cosubstrates than when it served as the sole carbon source. An approach to the evaluation of the specific transformation rate of fluorene during batch cultivations is proposed. PMID- 12751244 TI - [The study of Erwinia carotovora bacteriocins in the nalidixic acid resistant Erwinia carotovora]. AB - A novel approach is proposed for the study of the macromolecular bacteriocins of Erwinia carotovora (MCTVs). The approach lies in that the bacteriocinogeny of pectolytic erwinia is studied using a lawn of a bacterial mutant resistant to nalidixic acid, an inducer of MCTVs. The high efficiency of this approach was demonstrated by studying carotovoricins in 104 different E. carotovora strains, 88% of which bear MCTVs, distinguished by the morphology of zones of induced lysis on a lawn of susceptible cells, the lysis pattern, and some other characteristics. Preliminary studies by this approach showed that there is no correlation between the occurrence of MCTVs in particular E. carotovora strains and the habitat of the host plants from which these strains were isolated. There are grounds to believe that the approach proposed can also be used for investigating bacterial lysogeny. PMID- 12751245 TI - [Production of oil-processing compounds by microorganisms from the Daqing oil field, China]. AB - Twenty pure cultures isolated from formation waters of the Daqing oil field were studied with respect to their capacity to produce surface-active compounds in media with individual hydrocarbons, lower alcohols, and fatty acids. Aerobic saprotrophic bacteria belonging to the genera Bacillus, Brevibacillus, Rhodococcus, Dietzia, Kocuria, Gordonia, Cellulomonas, Clavibacter, Pseudomonas, and Acinetobacter decreased the surface tension of cultivation media from 55-63 to 28-44 mN/m. Strains of Bacillus cereus, Rhodococcus ruber, and Bacillus licheniformis produced biosurfactants most actively. Bacteria of the genera Rhodococcus, Dietzia, Kocuria, and Gordonia produced exopolysaccharides in media with hydrocarbons. Culture liquids of the strains of R. ruber and B. licheniformis exhibited oil-releasing effect. Thus, the Daqing oil field is inhabited by aerobic bacteria capable of producing effective oil-releasing agents. PMID- 12751246 TI - [Microorganisms in heat supply lines and internal corrosion of steel pipes]. AB - In laboratory experiments with batch cultures of thermophilic microorganisms isolated from urban heat supply systems, the growth of sulfate-reducing, iron oxidizing, and iron-reducing bacteria was found to accelerate the corrosion rate of the steel-3 plates used in the pipelines. In the absence of bacteria and dissolved oxygen, minimal, corrosion was determined. The aforementioned microorganisms, as well as sulfur-oxidizing bacteria, were found to be widespread in water and corrosion deposits in low-alloy steel pipelines (both delivery and return) of the Moscow heat networks, as well as in the corrosion deposits on the steel-3 plates in a testing unit supplied with the network water. The microorganisms were found in samples with water pH ranging from 8.1 to 9.6 and a temperature lower than 90 degrees C. Magnetite, lepidocrocite, goethite, X-ray amorphous ferric oxide were the corrosion products identified on the steel-3 plates, as well as siderite, aragonite, and S0. The effect of microbiological processes on the rate of electrochemical corrosion was evaluated from the accumulation of corrosion deposits and from variation in total and local corrosion of the steel plates in a testing unit. PMID- 12751247 TI - [The biometric analysis of bacteria in soil]. AB - The biometric analysis of bacterial cells in soil by light, fluorescence, and scanning electron microscopy showed that their average size is 0.8 micron in diameter, 1.4 microns in length, and 0.7 micron 3 in volume. In soil loci with enhanced microbiological activity (the rhizoplane of plants and the intestinal tract of soil invertebrates), the average size of bacterial cells was found to be 40% smaller than that of cells occurring in other parts of soil. It is the first experimental evidence showing that the metabolic activity of soil bacteria and their concentration and allometric parameters are related. PMID- 12751248 TI - [The structure and biogeochemical activity of the phototrophic communities from the Bol'sherechenskii alkaline hot spring]. AB - Microbial communities growing in the bed of the alkaline, sulfide hot spring Bol'sherechenskii (the Baikal rift area) were studied over many years (1986 2001). The effluent water temperature ranged from 72 to 74 degrees C, pH was from 9.25 to 9.8, and sulfide content was from 12 to 13.4 mg/ml. Simultaneous effects of several extreme factors restrict the spread of phototrophic microorganisms. Visible microbial fouling appears with a decrease in the temperature to 62 degrees C and in the sulfide content to 5.9 mg/l. Cyanobacteria predominated in all biological zones of the microbial mat. The filamentous cyanobacteria of the genus Phormidium are the major mat-forming organisms, whereas unicellular cyanobacteria and the filamentous green bacterium Chloroflexus aurantiacus are minor components of the phototrophic communities. No cyanobacteria of the species Mastigocladus laminosus, typical of neutral and subacid springs, were identified. Seventeen species of both anoxygenic phototrophic bacteria and cyanobacteria were isolated from the microbial mats, most of which exhibited optimum growth at 20 to 45 degrees C. The anoxygenic phototrophs were neutrophiles with pH optimum at about 7. The cyanobacteria were the most adapted to the alkaline conditions in the spring. Their optimum growth was observed at pH 8.5-9.0. As determined by the in situ radioisotope method, the optimal growth and decomposition rates were observed at 40-32 degrees C, which is 10 to 15 degrees C lower than the same parameter in the sulfide-deficient Octopus Spring (Yellowstone, United States). The maximum chlorophyll a concentration was 555 mg/m2 at 40 degrees C. Total rate of photosynthesis in the mats reached 1.3 g C/m2 per day. The maximum rate of dark fixation of carbon dioxide in the microbial mats was 0.806 g C/m2 per day. The maximum rate of sulfate reduction comprised 0.367 g S/m2 per day at 40 degrees C. The rate of methanogenesis did not exceed 1.188 micrograms C/m2 per day. The role of methanogenesis in the terminal decomposition of the organic matter was insignificant. Methane formation consumed 100 times less organic matter than sulfate reduction. PMID- 12751250 TI - [The distribution of bacterioplankton and mesozooplankton biomass in the White and Barents Seas coastal water]. AB - The total population density and the biomass of bacterioplankton, mesozooplankton, and phosphate-accumulating bacteria (PAB) were estimated during the 2000-2001 summer-autumn seasons in the coastal waters of the White and Barents Seas, which are subjects to the action of tidal and sea currents, the inflow of riverine waters, and anthropogenic impact. In the shallow estuarine waters with salinities of 6.5-32@1000 near the Chernaya, Pesha, and Pechora River mouths, the population of PAB fluctuated from 0.1 to 9.1 million cells/ml (0-36% of the total bacterial population). In pelagic seawaters, which are low in phosphates (12-50 micrograms/l) and are characterized by an increased iron/phosphorus ratio (2.0-3.6), bacterioplankton amounted to 0.1-1.6 million cells/ml and was mainly represented by small organisms with a volume of 0.08-0.15 micron 3, commonly lacking intracellular polyphosphates. In the pelagic zone of the Barents Sea, the biomass of mesozooplankton (Bz) was comparable with that of bacterioplankton (Bb = 39-175 mg/m3), the Bb/Bz ratio being 1.4-4.6. Off the Varandeiskii, Pechora, and Kolguyev oil terminals, Bb increased to 155-300 mg/m3 and the Bb/Bz ratio rose to 1.4 to 50.3 (with an average value of 20.9), presumably due to the severe anthropogenic impact on these waters. In this case, the dense population of bacterioplankton (0.9-7.6 million cells/ml) was mainly represented by large cells (0.12-0.76 micron 3 in volume), most of which (3-43% of the total bacterioplankton population) contained polyphosphates. The chemical composition of these waters was characterized by an elevated content of the total phosphorus (65-128 micrograms/l) and by a low iron/phosphorus ratio (0.9-1.2). PMID- 12751249 TI - [Microbial biodiversity in the Lake Baikal water]. AB - The investigation of the microbial community of Lake Baikal by the methods of general and molecular microbiology showed that culturable bacterial strains were represented by various known genera. The lake water contains a great number of bacterial morphotypes, as revealed by electron microscopy, and a great diversity of nonculturable microorganisms belonging to different phylogenetic groups, as revealed by 16S rRNA gene fragment sequencing. The inference is made that the microbial community of Lake Baikal contains not only the known species but also new, possibly endemic to the lake, bacterial species. PMID- 12751251 TI - [Microbial metabolism of the carbon and sulfur cycles in Shira Lake (Khakasia)]. AB - Microbiological and biogeochemical studies of the meromictic saline Lake Shira (Khakasia) were conducted. In the upper part of the hydrogen-sulfide zone, at a depth of 13.5-14 m, there was a pale pink layer of water due to the development of purple bacteria (6 x 10(5) cells/ml), which were assigned by their morphological and spectral characteristics to Lamprocystis purpureus (formerly Amoebobacter purpurea). In August, the production of organic matter (OM) in Lake Shira was estimated to be 943 mg C/(m2 day). The contribution of anoxygenic photosynthesis was insignificant (about 7% of the total OM production). The share of bacterial chemosynthesis was still less (no more than 2%). In the anaerobic zone, the community of sulfate-reducing bacteria played a decisive role in the terminal decomposition of OM. The maximal rates of sulfate reduction were observed in the near-bottom water (114 micrograms S/(1 day)) and in the surface layer of bottom sediments (901 micrograms S/(dm3 day)). The daily expenditure of Corg for sulfate reduction was 73% of Corg formed daily in the processes of oxygenic and anoxygenic photosynthesis and bacterial chemosynthesis. The profile of methane distribution in the water column and bottom sediments was typical of meromictic reservoirs. The methane content in the water column increased beginning with the thermocline (7-8 m), and reached maximum values in the near bottom water (17 microliters/l). In bottom sediments, the greatest methane concentrations (57 microliters/l) were observed in the surface layer (0-3 cm). The integral rate of methane formation in the water column and bottom sediments was almost an order of magnitude higher than the rate of its oxidation by aerobic and anaerobic methanotrophic microorganisms. PMID- 12751252 TI - [Seed bacterization and rhizosphere of wheat seedlings colonization by Bacillus Cohn]. AB - The dynamics of introduced antagonistic bacteria in the spring wheat rhizosphere was studied in small-plot field experiments during several growing seasons. The population density of introduced bacteria was found to considerably depend on the inoculum dose. At sufficiently high inoculum doses, the introduced bacteria remained in the wheat rhizosphere over the entire vegetative period (88-109 days). The maximum population density of introduced bacteria was observed in the early terms of plant development. No correlation was found between the population density of introduced bacteria and the degree of suppression of root rot or the structural crop yield parameters. The beneficial effect of preplanting seed bacterization on wheat plants was, as a rule, profound only during unfavorable growing seasons. PMID- 12751253 TI - [The effect of sulfur on the hydrocarbon-oxidizing bacteria of different genera growth]. PMID- 12751254 TI - [Morphological and physiological differences between fast- and slow-growing Escherichia coli]. PMID- 12751255 TI - [Degradation of 3,4-dichloroaniline by Pseudomonas fluorescens 26-K]. PMID- 12751256 TI - [Study of the aquatic bacterial community composition of Baikal lake by in situ hybridization assay]. PMID- 12751257 TI - [Role of the heterotrophic bacteria associated with the cyanobacterium Nostoc muscorum in cadmium sulfide synthesis]. PMID- 12751258 TI - Histoplasmosis. AB - The diagnosis of histoplasmosis depends on recognition of the different clinical manifestations of infection, accompanied by knowledge of the accuracy and limitations of tests used for diagnosis of fungal infections. Although most infections are asymptomatic, histoplasmosis can be severe, or even fatal, in patients who have experienced a heavy exposure, have underlying immune defects, or develop progressive disease that is not recognized and treated. This article provides an up-to-date clinical review of histoplasmosis, focusing on recognition, diagnosis, and management. PMID- 12751259 TI - Blastomycosis. AB - Blastomycosis is an endemic mycoses in the central United States caused by a dimorphic fungus, Blastomyces dermatitidis, that exists in nature in mycelial phase and converts to yeast phase at body temperature. The organism may produce epidemics of infection following a point source of infection or sporadic endemic infection. Blastomycosis can be a subclinical illness with subsequent protection against progressive infection afforded by cellular immune mechanisms, but it may present with progressive disease with either pulmonary or extrapulmonary disease or both. Itraconazole has been shown to be the drug of choice for both infections, except in cases of life-threatening infection when amphotericin B should be used. PMID- 12751260 TI - Coccidioidomycosis. AB - Coccidioides, a fungus, is endemic to specific parts of the Western Hemisphere. This article examines the prevalence, pathogenesis and host defense, clinical manifestations, diagnosis, and treatment of coccidioidomycosis. PMID- 12751261 TI - Subcutaneous mycoses. AB - Subcutaneous mycoses include a heterogeneous group of fungal infections that develop at the site of transcutaneous trauma. Infection slowly evolves as the etiologic agent survives and adapts to the adverse host tissue environment. Diagnosis rests on clinical presentation, histopathology, and culture of the etiologic agents. This article considers sporotrichosis, chromoblastomycosis, and mycetoma. PMID- 12751262 TI - Cutaneous infections dermatophytosis, onychomycosis, and tinea versicolor. AB - Cutaneous fungal infections cause significant morbidity for healthy and ill patients. The incidence of some dermatomycoses is increasing, despite availability of newer and better topical and systemic treatments. Fungal remnants last months to years under the ideal conditions, allowing continued spread of infection. Mycoses treated in one area may recur because of organism travel from concomitant areas of infection. Failure of patients and physicians to recognize a fungal etiology early may lead to more extensive, severe, or difficult-to-treat infections. Finally, a patient's concurrent illnesses may play a part in susceptibility and ability to manage fungal infections. For these reasons, scientists have studied and developed newer antifungal agents with better efficacy and greater convenience in dosing. These drugs, however, still have side effects and medication interactions that may limit their use in some patients. Better efforts to educate patients and physicians alike may aid in faster recognition and treatment of dermatophytoses. More research is needed to continue to develop drugs suitable for use in a broader range of patients and diagnostic tests that may be quicker or more specific than conventional ones. PMID- 12751263 TI - Fungal infections in the recipients of solid organ transplantation. AB - The advent of effective antibacterial and antiviral prophylatic and therapeutic strategies has led to the emergence of opportunistic mycoses as a principal cause of infection-related mortality in organ transplant recipients. Candida and Aspergillus species have accounted for most invasive fungal infections in organ transplant recipients. Epidemiologic trends within the last decade, however, are notable for the emergence of mycelial fungi other than Aspergillus as increasingly important pathogens in these patients. This article reviews the epidemiology, clinical manifestations, pathogenetic basis, diagnosis, and management of invasive fungal infections after organ transplantation in context of emerging trends and new developments in these areas. PMID- 12751264 TI - Laboratory diagnosis of invasive mycoses. AB - Rising numbers of immunocompromised patients have led to an ever-increasing population at risk of invasive fungal disease. Much has been achieved in the laboratory diagnosis of these infections, such as advances in blood culture systems, and the development of new biochemical, antigen detection assays, and molecular methodologies. More standardized susceptibility testing guidelines provide for better therapeutic interventions. In an era of economic cutbacks in health care, future challenges include the development of cost-effective and technically simplified systems, which provide early detection and identification of common and emerging fungal pathogens. It will, however, take some time to establish the clinical relevance of these new methodologies in different patient populations. PMID- 12751265 TI - Clinical pharmacology of antifungal compounds. AB - Prompted by the worldwide surge in fungal infections, the past decade has witnessed a considerable expansion in antifungal drug research. New compounds have entered the clinical arena, and major progress has been made in defining paradigms of antifungal therapies. This article provides an up-to-date review on the clinical pharmacology, indications, and dosage recommendations of approved and currently investigational therapeutics for treatment of invasive fungal infections in adult and pediatric patients. PMID- 12751266 TI - Immunomodulation of invasive fungal infections. AB - Genetic and acquired (disease- or therapy- related) host immune factors increase the risk for IFIs. In addition to antifungal drug therapy, modulation of host defenses by the use of HGFs and IFN-gamma has been supported by extensive in vitro and in vivo preclinical data. Clinical studies on the prevention or the adjunctive therapy of IFIs in combination with antifungal agents are limited, however, and do not allow specific recommendations for their cost-effective use in most of the immunodeficient settings. There is an urgent need to push forward with well-structured, randomized clinical trials to determine optimal dose, duration, and timing for different combinations of immunotherapy and antifungal agents in high-risk patients. PMID- 12751267 TI - The conduct of in vitro and in vivo drug-drug interaction studies: a PhRMA perspective. AB - Current regulatory guidances do not address specific study designs for in vitro and in vivo drug-drug interaction studies. There is a common desire by regulatory authorities and by industry sponsors to harmonize approaches to allow for a better assessment of the significance of findings across different studies and drugs. There is also a growing consensus for the standardization of cytochrome P450 (CYP) probe substrates, inhibitors, and inducers and for the development of classification systems to improve the communication of risk to health care providers and patients. While existing guidances cover mainly CYP-mediated drug interactions, the importance of other mechanisms, such as transporters, has been recognized more recently and should also be addressed. This paper was prepared by the Pharmaceutical Research and Manufacturers of America (PhRMA) Drug Metabolism and Clinical Pharmacology Technical Working Groups and represents the current industry position. The intent is to define a minimal best practice for in vitro and in vivo pharmacokinetic drug-drug interaction studies targeted to development (not discovery support) and to define a data package that can be expected by regulatory agencies in compound registration dossiers. PMID- 12751268 TI - Evaluation of the cohort size in phase I dose escalation trials based on laboratory data. AB - A survey of Phase I dose escalation trials published since 1995 shows that there is great disparity in all aspects of the design of the studies, and the cohort sizes range from 2 to 16 subjects with a great variety in the distribution between active and placebo-treated subjects. This study investigates the impact of the cohort size on Type I error and power in Phase I dose escalation trials based on laboratory data, with the hospitalization-induced increase in hepatic enzyme levels taken into consideration. The power of a Phase I dose escalation trial is very low, and only events with a very high probability of occurrence are detectable with acceptable power. For studies with cohort sizes smaller than 6 active subjects, there is much to gain with the inclusion of 1 extra subject, but for more than 10 subjects, little is gained by increasing the cohort size. With increasing cohort sizes, the probability of spontaneous non-drug-related events also increases, and this background rate needs to be considered when evaluating the trial. PMID- 12751269 TI - Ensuring supplies of quality diethylcarbamazine citrate (DEC). AB - In 1997, the World Health Organization (WHO) made a commitment to eliminate lymphatic filariasis. The WHO Global Program to Eliminate Lymphatic Filariasis (WHO-FIL) needed a reliable supply of diethylcarbamazine citrate (DEC) of known acceptable quality at an affordable price, so in August 1999, it started the DEC Project. Today's standards required development of a modern stability-indicating assay method for DEC and for DEC tablet dissolution. ADD Advanced Drug Delivery Technologies (Switzerland) developed a high-pressure liquid chromatography assay, which was independently validated and is in the United States Pharmacopoeia 25 (2002). After a global search, the project found that almost all existing and potential DEC active pharmaceutical ingredient (API) and tablet manufacturers are in low-income countries. The project constructed an audit team to conduct on-site audits to assess good manufacturing practices according to European Union standards. National/state inspectors accompany the audit team. The team prequalified one DEC API manufacturer and three DEC tablet manufacturers. The project plans to increase the number of prequalified DEC manufacturers. Now, WHO FIL only purchases from prequalified manufacturers. Consolidation of several national program DEC requirements into a limited international competitive bid reduced the price for DEC tablets between 30% and 45%, compared to previous small scale WHO purchases. PMID- 12751270 TI - Clinical pharmacology of topiramate versus lamotrigine versus phenobarbital: comparison of efficacy and side effects using odds ratios. AB - Clinical pharmacologists, neurologists, internists, and all health care givers must consider the efficacy, safety, and side effect profile of a given antiepileptic drug (AED) when determining which drug is best for a given patient. The first purpose of this paper is to address whether the "new" AEDs have advantages over the "old" drugs. The second purpose is to teach those interested in clinical pharmacology about the use of Web-based information access to answer a neurology/clinical pharmacology problem: to compare the efficacy and side effects of topiramate versus lamotrigine versus phenobarbital using odds ratios. Cost of all three AEDs was also compared. A number of new AEDs, including topiramate and lamotrigine, have been developed for chronic focal and secondarily generalized epileptic seizures. Efficacy of these drugs as anticonvulsants does not seem to be superior to that of traditional anticonvulsants such as phenobarbital. However, the advantage of the new drugs is a different spectrum of possible adverse events. Newer AEDs may or may not induce sedation and may minimize noncompliance by reducing side effects of lethargy and cognitive impairment. The difficulty in achieving therapeutic dosage because of side effects makes one consider whether these agents are "better" than the oldest and most side effect-prone AED, phenobarbital. The new AEDs have less frequent interactions, leading to improved tolerability with comedication. This exercise compares two "new" AEDs, topiramate and lamotrigine, with phenobarbital by evaluating efficacies and side effects using relative odds ratios, a method commonly used in drug development research. Development of new algorithms and/or new knowledge will bring beneficial tools to all clinical pharmacologists. PMID- 12751271 TI - The COX-2 selective inhibitor, valdecoxib, does not impair platelet function in the elderly: results of a randomized controlled trial. AB - The effects of the new cyclooxygenase (COX)-2 selective inhibitor, valdecoxib (40 mg bid; n = 17), on platelet function were evaluated, along with ibuprofen (800 mg tid; n = 15) and placebo (n = 15), in healthy elderly subjects (65-85 years) in this 7.5-day, randomized, single-center, double-blind study. Platelet aggregation (to sodium arachidonate, collagen, and adenosine diphosphate), bleeding time, and serum thromboxane B2 (TxB2) concentrations were measured up to 8 hours postdose on Days 1 and 8. Valdecoxib had no platelet effects, while ibuprofen significantly decreased platelet aggregation, significantly increased bleeding time (2-4 h postdose on each day), and significantly decreased TxB2 levels at all time points. In conclusion, unlike ibuprofen, valdecoxib 40 mg bid spares platelet COX-1 function in healthy elderly subjects. Valdecoxib's lack of effect on platelet aggregation and bleeding time suggests that it will have an improved clinical profile over nonselective NSAIDs, particularly in patients for whom bleeding complications are a concern. PMID- 12751272 TI - Galantamine population pharmacokinetics in patients with Alzheimer's disease: modeling and simulations. AB - Galantamine is a reversible, competitive inhibitor of acetylcholinesterase and an allosteric modulator of nicotinic acetylcholine receptors. It is cleared by renal and hepatic mechanisms, including metabolism by the CYP 450 2D6 and 3A4 isoenzymes. The authors estimated the population pharmacokinetics of galantamine using nonlinear mixed-effects modeling as implemented in NONMEM software. Data from 15 clinical studies (1089 individuals, 7480 concentration measurements in total) were used to examine the effect of body size, demographic characteristics, and concomitant disease status on galantamine pharmacokinetic parameters. Galantamine clearance was shown to decrease with age and increase with body weight and creatinine clearance of individuals. Median clearance in male and female patients with Alzheimer's disease (AD) was 14.8 and 12.4 L/h, respectively. The dissimilarity was related to the body weight difference, not to the real gender effect. Metabolic clearance was reduced by 60% in patients with moderate or severe hepatic dysfunction (Pugh score 7 or higher). Simulations were performed to assess the impact of hepatic impairment and renal insufficiency on peak plasma concentration of galantamine. Simulations confirmed the need for slower dose titration in patients with hepatic impairment: 4 mg daily during 1 week followed by 4, 8, and 12 mg bid, with each dose level during 1 week compared to the standard titration scheme 4-8-12-16 mg bid. However, no significant differences between plasma levels in AD patients with and without severe renal insufficiency were found. CYP 450 2D6 genotype also influenced galantamine clearance but not to the extent that dose adjustment is required. PMID- 12751273 TI - Effect of unilateral nephrectomy on the pharmacokinetics of atenolol in humans. AB - Unilateral nephrectomy is a common surgical procedure and involves functional adaptation of the remaining kidney. There is a paucity of data addressing whether drugs eliminated predominantly through the urinary tract, such as atenolol, can be effectively handled by unilaterally nephrectomized patients. This study was carried out in 28 subjects 1 month (10 subjects) and 1 year (10 subjects) after unilateral nephrectomy, as well as in 8 healthy volunteers who were administered atenolol as a single oral dose of 100 mg. Pharmacokinetic parameters of atenolol were calculated using a one-compartment open model. It was revealed that atenolol blood serum concentrations were significantly elevated in nephrectomized patients as compared with the control subjects, especially 1 month after the surgery. Patients nephrectomized 1 month prior to the study were characterized by an increase in AUC by 43% (p < 0.05), reduction of lambda z by 36% (p < 0.001) and CLT by 27% (p < 0.001), and prolongation of t1/2 by 60% (p < 0.001) in comparison with healthy volunteers. The values of atenolol pharmacokinetic parameters in patients 1 year after unilateral nephrectomy did not differ significantly as compared to the controls. However, a trend indicating an impaired elimination was noted. A decrease in lambda z by 16%, prolongation of t1/2 by 32%, and reduction of CLT by 32% were observed. The results suggest that unilateral nephrectomy impairs elimination of atenolol and possibly other drugs predominantly eliminated via the kidney. PMID- 12751274 TI - Pharmacoscintigraphic assessment of the regional drug absorption of the dual angiotensin-converting enzyme/neutral endopeptidase inhibitor, M100240, in healthy volunteers. AB - M100240 is the thioester of MDL 100,173, a dual angiotensin-converting enzyme (ACE)/neutral endopeptidase (NEP) inhibitor currently in phase II development. The purpose of this study was to evaluate the relative bioavaibility of M100240 in various regions of the gastrointestinal tract using the Enterion capsule, a noninvasive radiocontrolled device providing targeted drug delivery, to explore the absorption characteristics of M100240 in healthy volunteers. In addition, the absolute bioavailability of an immediate-release formulation of M100240 was assessed. Pharmacokinetic data were obtained from 13 healthy subjects in an open label, single-dose, randomized, five-period crossover study. Treatments included 25 mg M100240 administered via short intravenous infusion, oral immediate-release tablet administration, and oral Enterion capsule delivery of drug substance to the proximal small bowel, distal small bowel, and ascending colon. Each treatment was separated by a 14-day drug-free washout period. The localization of the Enterion capsule in the gastrointestinal tract was monitored using scintigraphic imaging. M100240 and MDL 100,173 plasma concentrations were quantified using a validated LC/MS/MS method, and pharmacokinetic parameters were calculated using noncompartmental methods. The estimates of relative bioavailability in the proximal small bowel, distal small bowel, and ascending colon relative to the oral immediate-release tablet are approximately 94%, 97%, and 41%, respectively. M100240 is primarily absorbed throughout the proximal and distal small bowel with modest absorption in the ascending colon. The absolute bioavailability estimate of the M100240 immediate-release formulation is 49%. These data characterize the fundamental in vivo performance attributes of M100240, thereby providing an approach for optimizing prototype modified-release formulations for this compound. PMID- 12751275 TI - Lack of bioequivalence when levofloxacin and calcium-fortified orange juice are coadministered to healthy volunteers. AB - Chelation interactions between drugs/supplements that contain large amounts of multivalent ions and the fluoroquinolones have been known for quite some time. However, there has been a lack of taking this interaction into account when they may be coadministered with foods that have been fortified with amounts of multiple multivalent ions that equal or exceed many supplement products. A previous study demonstrated that 12 ounces of calcium-fortified orange juice significantly decreased the bioequivalence of a dose of ciprofloxacin. This study examined, in 16 healthy volunteers, whether 12 ounces of orange juice with and without calcium fortification would demonstrate the same chelation interaction with single doses of levofloxacin. The results of the study demonstrated that both types of juice decreased levofloxacin Cmax values by 14% to 18% and prolonged tmax values by approximately 50%, with calcium-fortified orange juice decreasing Cmax enough to lose bioequivalence as compared to the control arm (89% [78.1%, 99.8%]). Due to the lack of change in overall exposure, it is thought that rather than a chelation interaction, levofloxacin and components of the orange juices competed for intestinal transport mechanisms such as P-glycoprotein and organic anion-transporting polypeptides, which resulted in the discovered interaction. These results further confirm the need to adjust regulatory studies to include bioequivalence/bioavailability studies that contain fortified foods more than high-calorie/high-fat foods to better reflect current American consumption habits. PMID- 12751276 TI - Isoniazid-induced anaphylaxis. AB - Anaphylaxis from isoniazid is a possible side effect to this commonly prescribed antibiotic. The case reported here illustrates that clinicians should be aware of this potentially fatal drug reaction. PMID- 12751277 TI - Strain-rate dependent stiffness of articular cartilage in unconfined compression. AB - The stiffness of articular cartilage is a nonlinear function of the strain amplitude and strain rate as well as the loading history, as a consequence of the flow of interstitial water and the stiffening of the collagen fibril network. This paper presents a full investigation of the interplay between the fluid kinetics and fibril stiffening of unconfined cartilage disks by analyzing over 200 cases with diverse material properties. The lower and upper elastic limits of the stress (under a given strain) are uniquely established by the instantaneous and equilibrium stiffness (obtained numerically for finite deformations and analytically for small deformations). These limits could be used to determine safe loading protocols in order that the stress in each solid constituent remains within its own elastic limit. For a given compressive strain applied at a low rate, the loading is close to the lower limit and is mostly borne directly by the solid constituents (with little contribution from the fluid). In contrast, however in case of faster compression, the extra loading is predominantly transported to the fibrillar matrix via rising fluid pressure with little increase of stress in the nonfibrillar matrix. The fibrillar matrix absorbs the loading increment by self-stiffening: the quicker the loading the faster the fibril stiffening until the upper elastic loading limit is reached. This self protective mechanism prevents cartilage from damage since the fibrils are strong in tension. The present work demonstrates the ability of the fibril reinfored poroelastic models to describe the strain rate dependent behavior of articular cartilage in unconfined compression using a mechanism of fibril stiffening mainly induced by the fluid flow. PMID- 12751278 TI - A growth mixture theory for cartilage with application to growth-related experiments on cartilage explants. AB - In this paper, we present a growth mixture model for cartilage. The main features of this model are illustrated in a simple equilibrium boundary-value problem that is chosen to illustrate how a mechanical theory of cartilage growth may be applied to growth-related experiments on cartilage explants. The cartilage growth mixture model describes the independent growth of the proteoglycan and collagen constituents due to volumetric mass deposition, which leads to the remodeling of the composition and the mechanical properties of the solid matrix. The model developed here also describes how the material constants of the collagen constituent depend on a scalar parameter that may change over time (e.g., crosslink density); this leads to a remodeling of the structural and mechanical properties of the collagen constituent. The equilibrium boundary-value problem that describes the changes observed in cartilage explants harvested at different stages of a growth or a degenerative process is formulated. This boundary-value problem is solved using existing experimental data for developing bovine cartilage explants harvested at three developmental stages. The solution of the boundary-value problem in conjunction with existing experimental data suggest the types of experimental studies that need to be conducted in the future to determine model parameters and to further refine the model. PMID- 12751279 TI - Toward an MRI-based method to measure non-uniform cartilage deformation: an MRI cyclic loading apparatus system and steady-state cyclic displacement of articular cartilage under compressive loading. AB - Recent magnetic resonance imaging (MRI) techniques have shown potential for measuring non-uniform deformations throughout the volume (i.e. three-dimensional (3D) deformations) in small orthopedic tissues such as articular cartilage. However, to analyze cartilage deformation using MRI techniques, a system is required which can construct images from multiple acquisitions of MRI signals from the cartilage in both the underformed and deformed states. The objectives of the work reported in this article were to 1) design an apparatus that could apply highly repeatable cyclic compressive loads of 400 N and operate in the bore of an MRI scanner, 2) demonstrate that the apparatus and MRI scanner can be successfully integrated to observe 3D deformations in a phantom material, 3) use the apparatus to determine the load cycle necessary to achieve a steady-state deformation response in normal bovine articular cartilage samples using a flat surfaced and nonporous indentor in unconfined compression. Composed of electronic and pneumatic components, the apparatus regulated pressure to a double-acting pneumatic cylinder so that (1) load-controlled compression cycles were applied to cartilage samples immersed in a saline bath, (2) loading and recovery periods within a cycle varied in time duration, and (3) load magnitude varied so that the stress applied to cartilage samples was within typical physiological ranges. In addition the apparatus allowed gating for MR image acquisition, and operation within the bore of an MRI scanner without creating image artifacts. The apparatus demonstrated high repeatability in load application with a standard deviation of 1.8% of the mean 400 N load applied. When the apparatus was integrated with an MRI scanner programmed with appropriate pulse sequences, images of a phantom material in both the underformed and deformed states were constructed by assembling data acquired through multiple signal acquisitions. Additionally, the number of cycles to reach a steady-state response in normal bovine articular cartilage was 49 for a total cycle duration of 5 seconds, but decreased to 33 and 27 for increasing total cycle durations of 10 and 15 seconds, respectively. Once the steady-state response was achieved, 95% of all displacements were within +/- 7.42 microns of the mean displacement, indicating that the displacement response to the cyclic loads was highly repeatable. With this performance, the MRI-loading apparatus system meets the requirements to create images of articular cartilage from which 3D deformation can be determined. PMID- 12751281 TI - Targeted drug aerosol deposition analysis for a four-generation lung airway model with hemispherical tumors. AB - One important research area of broad interest is the development of highly efficient drug delivery systems for desired site deposition and uptake. For example, controlled drug aerosol release and targeting to specific regions of the lung is a novel way to combat lung diseases, diabetes, virus infections, cancers, etc. Determination of feasible air-particle streams is a prerequisite for the development of such delivery devices, say, smart inhalers. The concept of "controlled particle release and targeting" is introduced and results are discussed for a representative model of bronchial lung airways afflicted with hemispherical tumors of different sizes and locations. It is shown that under normal particle inlet conditions a particle mass fraction of only up to 11% may deposit on the surface of a specific tumor with critical radius r/R approximately 1.25, while a controlled particle release achieves deposition fractions of 35 to 92% for a realistic combination of inlet Stokes and Reynolds numbers, depending mainly on tumor size. Furthermore, with the controlled release and targeting approach nearby healthy tissue is hardly impacted by the typically aggressive drug aerosols. Assuming laminar, quasi-steady, three-dimensional air flow and spherical non-interacting micron-particles in sequentially bifurcating rigid airways, the results were obtained using a validated commercial finite-volume code with user-enhanced programs on a high-end engineering workstation. The new concept is generic and hence should be applicable to other regions of the respiratory system as well. PMID- 12751280 TI - Analysis of hemodynamic fluid phase mass transport in a separated flow region. AB - The mass transfer behavior in the recirculation region downstream of an axisymmetric sudden expansion was examined. The Reynolds number, 500, and Schmidt number, 3200, were selected to model the mass transfer of molecules, such as ADP, in the arterial system. In a first step the transient mass transport applying zero diffusive flux at the wall was analyzed using experiments and two computational codes. The two codes were FLUENT, a commercially available finite volume method, and FTSP, a finite element code developed at Graz University of Technology. The comparison of the transient wall concentration values determined by the three methods was excellent and provides a measure of confidence for computational mass transfer calculations in convection dominated, separated flows. In a second step the effect of the flow separation on the stationary mass transport applying a permeability boundary condition at the water-permeable wall was analyzed using the finite element code FTSP. The results show an increase of luminal ADP surface concentration in the upstream and in the downstream tube of the sudden expansion geometry in the range of six and twelve percent of the bulk flow concentration. The effect of flow separation in the downstream tube on the wall concentration is a decrease of about ten percent of the difference between wall concentration and bulk concentration occurring at nearly fully developed flow at the downstream region at a distance of 66 downstream tube diameters from the expansion. The decrease of ADP flux into the wall is in the range of three percent of the flux at the downstream region. PMID- 12751282 TI - The effect of asymmetry in abdominal aortic aneurysms under physiologically realistic pulsatile flow conditions. AB - In the abdominal segment of the human aorta under a patient's average resting conditions, pulsatile blood flow exhibits complex laminar patterns with secondary flows induced by adjacent branches and irregular vessel geometries. The flow dynamics becomes more complex when there is a pathological condition that causes changes in the normal structural composition of the vessel wall, for example, in the presence of an aneurysm. This work examines the hemodynamics of pulsatile blood flow in hypothetical three-dimensional models of abdominal aortic aneurysms (AAAs). Numerical predictions of blood flow patterns and hemodynamic stresses in AAAs are performed in single-aneurysm, asymmetric, rigid wall models using the finite element method. We characterize pulsatile flow dynamics in AAAs for average resting conditions by means of identifying regions of disturbed flow and quantifying the disturbance by evaluating flow-induced stresses at the aneurysm wall, specifically wall pressure and wall shear stress. Physiologically realistic abdominal aortic blood flow is simulated under pulsatile conditions for the range of time-average Reynolds numbers 50 < or = Rem < or = 300, corresponding to a range of peak Reynolds numbers 262.5 < or = Repeak < or = 1575. The vortex dynamics induced by pulsatile flow in AAAs is depicted by a sequence of four different flow phases in one period of the cardiac pulse. Peak wall shear stress and peak wall pressure are reported as a function of the time-average Reynolds number and aneurysm asymmetry. The effect of asymmetry in hypothetically shaped AAAs is to increase the maximum wall shear stress at peak flow and to induce the appearance of secondary flows in late diastole. PMID- 12751283 TI - Accuracy and reproducibility of CFD predicted wall shear stress using 3D ultrasound images. AB - Computational fluid dynamics (CFD) flow simulation techniques have the potential to enhance our understanding of how haemodynamic factors are involved in atherosclerosis. Recently, 3D ultrasound has emerged as an alternative to other 3D imaging techniques, such as magnetic resonance angiography (MRA). The method can be used to generate realistic vascular geometry suitable for CFD simulations. In order to assess accuracy and reproducibility of the procedure from image acquisition to reconstruction to CFD simulation, a human carotid artery bifurcation phantom was scanned three times using 3D ultrasound. The geometry was reconstructed and flow simulations were carried out on the three sets as well as on a model generated using computer aided design (CAD) from the geometric information given by the manufacturer. It was found that the three reconstructed sets showed good reproducibility as well as satisfactory quantitative agreement with the CAD model. Analyzing two selected locations probably representing the 'worst cases,' accuracy comparing ultrasound and CAD reconstructed models was estimated to be between 7.2% and 7.7% of the maximum instantaneous WSS and reproducibility comparing the three scans to be between 8.2% and 10.7% of their average maximum. PMID- 12751284 TI - Modeling the human body/seat system in a vibration environment. AB - The vibration environment is a common man-made artificial surrounding with which humans have a limited tolerance to cope due to their body dynamics. This research studied the dynamic characteristics of a seated human body/seat system in a vibration environment. The main result is a multi degrees of freedom lumped parameter model that synthesizes two basic dynamics: (i) global human dynamics, the apparent mass phenomenon, including a systematic set of the model parameters for simulating various conditions like body posture, backrest, footrest, muscle tension, and vibration directions, and (ii) the local human dynamics, represented by the human pelvis/vibrating seat contact, using a cushioning interface. The model and its selected parameters successfully described the main effects of the apparent mass phenomenon compared to experimental data documented in the literature. The model provided an analytical tool for human body dynamics research. It also enabled a primary tool for seat and cushioning design. The model was further used to develop design guidelines for a composite cushion using the principle of quasi-uniform body/seat contact force distribution. In terms of evenly distributing the contact forces, the best result for the different materials and cushion geometries simulated in the current study was achieved using a two layer shaped geometry cushion built from three materials. Combining the geometry and the mechanical characteristics of a structure under large deformation into a lumped parameter model enables successful analysis of the human/seat interface system and provides practical results for body protection in dynamic environment. PMID- 12751285 TI - A spatial mechanism with higher pairs for modelling the human knee joint. AB - By generalizing a previous model proposed in the literature, a new spatial kinematic model of the knee joint passive motion is presented. The model is based on an equivalent spatial parallel mechanism which relies upon the assumption that fibers within the anterior cruciate ligament (ACL), the medial collateral ligament (MCL) and the posterior cruciate ligament (PCL) can be considered as isometric during the knee flexion in passive motion (virtually unloaded motion). The articular surfaces of femoral and tibial condyles are modelled as 3-D surfaces of general shapes. In particular, the paper presents the closure equations of the new mechanism both for surfaces represented by means of scalar equations that have the Cartesian coordinates of the points of the surface as variables and for surfaces represented in parametric form. An example of simulation is presented in the case both femoral condyles are modelled as ellipsoidal surfaces and tibial condyles as spherical surfaces. The results of the simulation are compared to those of the previous models and to measurements. The comparison confirms the expectation that a better approximation of the tibiofemoral condyle surfaces leads to a more accurate model of the knee passive motion. PMID- 12751286 TI - In-vivo measurement of dynamic joint motion using high speed biplane radiography and CT: application to canine ACL deficiency. AB - Dynamic assessment of three-dimensional (3D) skeletal kinematics is essential for understanding normal joint function as well as the effects of injury or disease. This paper presents a novel technique for measuring in-vivo skeletal kinematics that combines data collected from high-speed biplane radiography and static computed tomography (CT). The goals of the present study were to demonstrate that highly precise measurements can be obtained during dynamic movement studies employing high frame-rate biplane video-radiography, to develop a method for expressing joint kinematics in an anatomically relevant coordinate system and to demonstrate the application of this technique by calculating canine tibio-femoral kinematics during dynamic motion. The method consists of four components: the generation and acquisition of high frame rate biplane radiographs, identification and 3D tracking of implanted bone markers, CT-based coordinate system determination, and kinematic analysis routines for determining joint motion in anatomically based coordinates. Results from dynamic tracking of markers inserted in a phantom object showed the system bias was insignificant (-0.02 mm). The average precision in tracking implanted markers in-vivo was 0.064 mm for the distance between markers and 0.31 degree for the angles between markers. Across trial standard deviations for tibio-femoral translations were similar for all three motion directions, averaging 0.14 mm (range 0.08 to 0.20 mm). Variability in tibio-femoral rotations was more dependent on rotation axis, with across-trial standard deviations averaging 1.71 degrees for flexion/extension, 0.90 degree for internal/external rotation, and 0.40 degree for varus/valgus rotation. Advantages of this technique over traditional motion analysis methods include the elimination of skin motion artifacts, improved tracking precision and the ability to present results in a consistent anatomical reference frame. PMID- 12751287 TI - The use of sequential MR image sets for determining tibiofemoral motion: reliability of coordinate systems and accuracy of motion tracking algorithm. AB - The use of magnetic resonance imaging has been proposed by many investigators for establishment of joint reference systems and kinematic tracking of musculoskeletal joints. In this study, the intraobserver and interobserver reliability of a strategy to establish anatomic reference systems using manually selected fiducial points were quantified for seven sets of MR images of the human knee joint. The standard error of the measurement of the intraobserver and interobserver errors were less than 2.6 degrees, and 1.2 mm for relative tibiofemoral orientation and displacement, respectively. An automated motion tracking algorithm was also validated with a controlled motion experiment in a cadaveric knee joint. The controlled displacements and rotations prescribed in our motion tracking validation were highly correlated to those predicted (Pearson's correlation = 0.99, RMS errors = 0.39 mm, 0.38 degree). Finally, the system for anatomic reference system definition and motion tracking was demonstrated with a set of MR images of in vivo passive flexion in the human knee. PMID- 12751288 TI - A biomechanical evaluation of whiplash using a multi-body dynamic model. AB - This paper presents a biomechanical evaluation of whiplash injury potential during the initial extension motion of the head in a rear-end collision. A four segment dynamic model is developed in the sagittal plane for the analysis. The model response is validated using the existing experimental data and is shown to simulate the "S-shape" kinematics of the cervical spine and the resulting dynamics observed in human and cadaver experiments. The model is then used to evaluate the effects of parameters such as collision severity, head/headrest separation, and the initial head orientation in the sagittal plane on the "S shape" kinematics of the cervical spine and the resulting neck loads. It is shown, for example, that the cervical spine forms an "S-shape" for a range of change in speeds and that at lower and higher speeds changes the spine does not form the "S-shape." Furthermore, it is shown that the "S-shape" formation also depends on the head to headrest separation distance. PMID- 12751289 TI - Morphology and stress-strain properties along the small intestine in the rat. AB - The stress-strain relationship is determined by the inherent mechanical properties of the intestinal wall, the geometric configurations, the loading conditions and the zero-stress state of the segment. The purpose of this project was to provide morphometric and biomechanical data for rat duodenum, jejunum and ileum. The circumferential strains were referenced to the zero-stress state. Large morphometric variations were found along the small intestine with an increase in the outer circumferential length and luminal area and a decrease in wall thickness in distal direction. The serosal residual strain was tensile and decreased in distal direction (P < 0.05). The mucosal residual strain was compressive and the absolute value decreased in distal direction (P < 0.001). The stress-strain experiments showed that the duodenum was stiffest. All segments were stiffest in longitudinal direction (P < 0.05). In conclusion, axial variation in morphometric and biomechanical properties was found in the small intestine. The zero-stress state must be considered in future biomechanical studies in the gastrointestinal tract. PMID- 12751290 TI - Constitutive modeling of porcine coronary arteries using designed experiments. AB - The development of new coronary artery constitutive models is of critical importance in the design and analysis of coronary replacement grafts. In this study, a two-parameter logarithmic complementary energy function, with normalized measured force and internal pressure as the independent variables and strains as the dependent variables, was developed for healthy porcine coronary arteries. Data was collected according to an experimental design with measured force ranging from 9.8 to 201 mN and internal pressure ranging from 0.1 to 16.1 kPa (1 to 121 mmHg). Comparisons of the estimated constitutive parameters showed statistically significant differences between the left anterior descending [LAD] and right coronary artery [RCA], but no differences between the LAD and left circumflex [LCX] or between the LCX and RCA. Point-by-point strain comparisons confirm the findings of the model parameter study and isolate the difference to the axial strain response. Average axial strains for the LAD, LCX, and RCA are 0.026 +/- 0.009, 0.015 +/- 0.005, and 0.011 +/- 0.009, respectively, at all physiologic loads, suggesting that the axial strains in the LAD are significantly higher than in the other regions. PMID- 12751292 TI - Axial mechanical properties of fresh human cerebral blood vessels. AB - Human cerebral blood vessels are frequently damaged in head impact, whether accidental or deliberate, resulting in intracranial bleeding. Additionally, the vasculature constitutes the support structure for the brain and, hence, plays a key role in the cranial load response. Quantification of its mechanical behavior, including limiting loads, is thus required for a proper understanding and modeling of traumatic brain injury--as well as providing substantial assistance in the development and application of preventive measures. It is believed that axial stretching is the dominant loading mode for the blood vessels, regardless of the nature of the insult. Eighteen arteries and fourteen veins were obtained from the cortical surface of the cerebral temporal lobe of patients undergoing surgery. These vessels were stretched to failure in the longitudinal direction, either quasi-statically or dynamically. The significance of specimen and experiment parameters was determined using multivariate analysis of variance (MANOVA) testing. Results demonstrate that the arteries were considerably stiffer than the veins, carrying approximately twice as much stress at failure but withstanding only half as much stretch. No significant rate dependence was measured over a strain rate range of more than four orders of magnitude (0.01 to 500 s -1). PMID- 12751291 TI - Incorporation of experimentally-derived fiber orientation into a structural constitutive model for planar collagenous tissues. AB - Structural constitutive models integrate information on tissue composition and structure, avoiding ambiguities in material characterization. However, critical structural information (such as fiber orientation) must be modeled using assumed statistical distributions, with the distribution parameters estimated from fits to the mechanical test data. Thus, full realization of structural approaches continues to be limited without direct quantitative structural information for direct implementation or to validate model predictions. In the present study, fiber orientation information obtained using small angle light scattering (SALS) was directly incorporated into a structural constitutive model based on work by Lanir (J. Biomech., v. 16, pp. 1-12, 1983). Demonstration of the model was performed using existing biaxial mechanical and fiber orientation data for native bovine pericardium (Sacks and Chuong, ABME, v.26, pp. 892-902, 1998). The structural constitutive model accurately predicted the complete measured biaxial mechanical response. An important aspect of this approach is that only a single equibiaxial test to determine the effective fiber stress-strain response and the SALS-derived fiber orientation distribution were required to determine the complete planar biaxial mechanical response. Changes in collagen fiber crimp under equibiaxial strain suggest that, at the meso-scale, fiber deformations follow the global tissue strains. This result supports the assumption of affine strain to estimate the fiber strains. However, future evaluations will have to be performed for tissue subjected to a wider range of strain to more fully validate the current approach. PMID- 12751293 TI - Determination of optimal graft lengths for posterior cruciate ligament reconstruction--a theoretical analysis. AB - Various graft materials have been used in posterior cruciate ligament (PCL) reconstruction. However, it is unclear if these grafts can reproduce the structural behavior of the PCL. This paper analyzed the effect of graft length on the structural behavior of the graft using a minimal deformation energy method. An analytical solution was obtained to determine the optimal effective graft length that can best reproduce the structural behavior of the PCL. This optimal graft length was determined as a function of the axial rigidity of the graft. Two typical grafts, bone-patella tendon-bone (BPTB) and Achilles tendon, were analyzed. The data demonstrated that in order to reproduce the PCL behavior, the effective length of a BPTB graft (10 mm width) should be 34 mm, while the Achilles tendon graft (with a cross sectional area of 55 mn2) needs to be 48 mm in length. Longer grafts result in less resistance and shorter graft increased the graft resistance. An initial graft tension cannot help recreate the overall structural behavior of the PCL. These results suggest that graft length is an important surgical variable in PCL reconstruction. An optimal reconstruction of the PCL should reproduce the structural properties of the PCL by using a graft with an optimal length. PMID- 12751295 TI - Building foundations. PMID- 12751294 TI - On a relationship between the Arrhenius parameters from thermal damage studies. PMID- 12751296 TI - Orbital penetration associated with tooth extraction. AB - Three cats and 2 dogs were evaluated for ophthalmologic complications associated with tooth extraction procedures. Orbital penetration leading to ocular and, in one case, brain trauma was secondary to iatrogenic injury from a dental elevator. Outcomes included enucleation of the affected eye in 3 cases, and death from brain abscessation in 1 case. Early treatment or, preferably, referral to a veterinary ophthalmology specialist may prevent such outcomes. Awareness of the anatomical proximity of caudal maxillary tooth roots and the orbit, appropriate interpretation of diagnostic intraoral dental radiographs, and technical proficiency in tooth extraction techniques will minimize these complications in veterinary dental practice. PMID- 12751297 TI - The effect of three types of rasps on the occlusal surface of equine cheek teeth: a scanning electron microscopic study. AB - Two hand rasps (tungsten chip blade, solid carbide blade) and an electrically driven solid carbide axial bur were used to treat the cheek teeth of 2 horses immediately postmortem. All teeth were normal and were rasped to a standard considered satisfactory in practice. Six teeth from each horse served as untreated controls. Following treatment, the teeth were extracted and the clinical crown removed and prepared for scanning electron microscopy. Teeth were also extracted and examined from a horse that had excessive dental treatment previously. Dental debris created by the procedures was collected and examined. All three rasp techniques resulted in amputation of odontoblast processes. The solid carbide blade cut deep gouges and grooves into the surface of the dentin, chipping the enamel and peripheral cement. No smear layer was created. Rasping with a tungsten chip blade created a partial smear layer and a smoother surface than the solid carbide blade. The electrically-driven bur produced a complete smear layer and removed all dental tissues to a smooth layer. The enamel had also been damaged by the electric bur. Crown particles collected after the procedures were larger following hand rasping compared with particles produced by the electric bur. The extent of damage to sensitive and vital dentin tissue was of concern. Further studies are required to establish the optimum technique for rasping equine cheek teeth. PMID- 12751298 TI - Maxillary dentigerous cyst in a cat. AB - A dentigerous cyst was diagnosed in the right rostral maxilla of a 6-month-old Siamese cat affecting the maxillary right canine tooth. The deciduous maxillary right canine tooth was extracted and the dentigerous cyst wall was curetted. The permanent tooth was not removed, however it was mobile following the surgical procedure. Examination 6-months following surgery indicated appropriate development of the tooth with no mobility or periodontal pocket formation. The maxillary right canine tooth had an abnormal shape but was functional with an acceptable appearance. PMID- 12751299 TI - Odontogenic periorbital fistula in a cat. PMID- 12751300 TI - Home care for the veterinary dental patient. AB - There is a dizzying array of home care products and aids available for veterinary clients to use for their pets. Although it can be confusing for veterinarians to determine which products to use and when, do not leave it up to your clients to decide for themselves. For those animals that need home care, take the time to discuss the situation with the owner and explain what they need to do. Listen to their thoughts and concerns, consider the animal involved, and then customize a program based on the individual situation. PMID- 12751301 TI - Surgical extraction of the maxillary canine tooth. PMID- 12751302 TI - Using the ISMP Medication Safety Self-Assessment to improve medication use processes. AB - BACKGROUND: The 194-item Institute for Safe Medication Practices (ISMP) Medication Safety Self-Assessment for hospitals is being used by a 21-hospital collaborative as a framework for understanding, evaluating, and systematically implementing medication use system safety improvements. METHODS: The results of the self-assessment were used to prioritize and organize the review of medication use processes, in which each self-assessment-defined "representative characteristic" is reviewed in detail, and "best implementation practices" for the characteristic are established. The collaborative concurrently identifies educational needs and develops tools to assist organizations in implementing improvements. RESULTS: By December 2002 participating organizations had implemented a wide variety of medication safety improvements. Collaborative member scores for the self-assessment increased approximately 20% during the initial assessment when the self-assessment was repeated by members in the second quarter of 2002. SUMMARY: Participant progress in improving medication safety practices is supportive of collaborative methods and the value of completing the ISMP self-assessment, expanding on the knowledge gained, developing effective implementation tools, and systematically applying lessons learned. PMID- 12751303 TI - Microsystems in health care: Part 4. Planning patient-centered care. AB - BACKGROUND: Clinical microsystems are the essential building blocks of all health systems. At the heart of an effective microsystem is a productive interaction between an informed, activated patient and a prepared, proactive practice staff. Support, which increases the patient's ability for self-management, is an essential result of a productive interaction. This series on high-performing clinical microsystems is based on interviews and site visits to 20 clinical microsystems in the United States. This fourth article in the series describes how high-performing microsystems design and plan patient-centered care. PLANNING PATIENT-CENTERED CARE: Well-planned, patient-centered care results in improved practice efficiency and better patient outcomes. However, planning this care is not an easy task. Excellent planned care requires that the microsystem have services that match what really matters to a patient and family and protected time to reflect and plan. Patient self-management support, clinical decision support, delivery system design, and clinical information systems must be planned to be effective, timely, and efficient for each individual patient and for all patients. CONCLUSION: Excellent planned services and planned care are attainable today in microsystems that understand what really matters to a patient and family and have the capacity to provide services to meet the patient's needs. PMID- 12751304 TI - Introducing practice-based learning and improvement ACGME core competencies into a family medicine residency curriculum. AB - BACKGROUND: The Accreditation Council for Graduate Medical Education (ACGME) recommends integrating improvement activities into residency training. A curricular change was designed at the Department of Family and Community Medicine, University of Louisville, to address selected ACGME competencies by incorporating practice-based improvement activities into the routine clinical work of family medicine residents. METHODS: Teams of residents, faculty, and office staff completed clinical improvement projects at three ambulatory care training sites. Residents were given academic credit for participation in team meetings. After 6 months, residents presented results to faculty, medical students, other residents, and staff from all three training sites. Residents, staff, and faculty were recognized for their participation. PROGRAM EVALUATION: Resident teams demonstrated ACGME competencies in practice-based improvement: Chart audits indicated improvement in clinical projects; quality improvement tools demonstrated analysis of root causes and understanding of the process; plan do-study-act cycle worksheets demonstrated the change process. CONCLUSIONS: Improvement activities that affect patient care and demonstrate selected ACGME competencies can be successfully incorporated into the daily work of family medicine residents. PMID- 12751306 TI - Role-playing case simulations: a tool to improve communication and enhance safety. PMID- 12751305 TI - Using a systemwide care path to enhance compliance with guidelines for acute myocardial infarction. AB - BACKGROUND: Several acute myocardial infarction (MI) guidelines and policy statements have been developed, but compliance rates are suboptimal. The cardiology section at Gundersen Lutheran Medical Center (La Crosse, Wisconsin) used a systemwide approach to enhance compliance with guidelines. METHODS AND RESULTS: Data were collected prospectively for a 4-year period (May 15, 1995-May 15, 1999) for all patients presenting with acute MI. In 1995 a multidisciplinary team developed protocols for each phase of MI management and designed clinical care paths with built-in accountability. The initiative resulted in improvements in all phases of acute MI care and met the benchmark recommendations in mean time to electrocardiogram, thrombolytic therapy, and aspirin and beta-blocker administration. Rates of prescriptions for secondary prevention were 92% for aspirin and beta-blocker and 97% for smoking cessation education at 4 years. SUMMARY: The care path for acute MI involved multiple disciplines and empowerment of nonspecialists and nonphysician practitioners during development and implementation, as well as continual education and retraining. The care path led to several improvements in performance scores. These findings indicate that the recommendations as set forth in the American College of Cardiology (ACC) and the American Heart Association (AHA) guidelines for managing acute MI are realistic and achievable, and they do not require additional resources. PMID- 12751307 TI - The pursuit of socially modifiable contingencies in mental health. AB - The effort to understand the meanings of the well-demonstrated linkages between mental health and one's locations in the social structure has commanded a great deal of research attention over the past half century. Following a brief consideration of my early efforts to make a contribution in this regard, some recent and ongoing work is summarized. In concert with important work by others, these findings support the conclusion that differences in exposure to social stress represent a much more critical contingency in mental health and substance use outcomes than has generally been assumed. In addition, reported results indicate that the lifetime experience of multiple adversities is quite common among young people in South Florida and, presumably, elsewhere and the likelihood that the compelling linkage observed between cumulative adversity and risk for psychiatric and substance disorders is causal in nature. It is suggested that the development of interventions in the service of stress prevention or reduction should command a greater proportion of the attention of researchers and interventionists. PMID- 12751308 TI - Does perceived discrimination affect health? Longitudinal relationships between work discrimination and women's physical and emotional health. AB - This study uses longitudinal data to examine the causal relationships between perceived work discrimination and women's physical and emotional health. Using data on 1,778 employed women in the National Longitudinal Survey of Mature Women, we investigate the structural and individual characteristics that predict later perceptions of discrimination and the effects of those perceptions on subsequent health. We find that perceptions of discrimination are influenced by job attitudes, prior experiences of discrimination, and work contexts, but prior health is not related to later perceptions. However, perceptions of discrimination do impact subsequent health, and these effects remain significant after controlling for prior emotional health, physical health limitations, discrimination, and job characteristics. Overall, the results provide even stronger support for the health impact of workplace discrimination and suggest a need for further longitudinal analyses of causes and consequences of perceived discrimination. PMID- 12751309 TI - Stressful neighborhoods and depression: a prospective study of the impact of neighborhood disorder. AB - Quantitative and qualitative research suggests that urban disadvantaged environments may be highly stressful to their inhabitants. Social disorganization may be deleterious to both physical and mental health. The relationships among perceptions of one's neighborhood, measures of social support and social integration, and level of subsequent depressive symptoms was examined with a community sample of 818 individuals screened for an HIV prevention intervention, most of whom were current or former drug users. After adjusting for baseline levels of depressive symptoms, perceptions of neighborhood characteristics (vandalism, litter or trash, vacant housing, teenagers hanging out, burglary, drug selling, and robbery) predicted depressive symptoms at a 9-month follow-up interview. Measures of social support and social integration, entered as interactions with neighborhood perceptions, did not buffer the effect of neighborhood perceptions. However, CES-D scores at follow-up for frequent church attendees were lower. The data support theories of social disorganization and social stress and suggest the need for structural intervention. PMID- 12751310 TI - Contraceptive self-efficacy: does it influence adolescents' contraceptive use? AB - This research investigates the relationship between contraceptive self-efficacy and contraceptive use, measured one year later, among adolescent boys and girls. Data are obtained from the two waves of the restricted use sample of the National Longitudinal Study of Adolescent Health (n = 3,577). Employing multiple regression and logistic regression strategies, we examine whether demographic and background characteristics influence contraceptive self-efficacy, and whether contraceptive self-efficacy increases the likelihood of contraceptive use. We find that adolescents who are female, older, live with step-parents, and whose mothers approve of contraceptive use report higher contraceptive self-efficacy, while adolescents whose mothers did not complete high school report lower contraceptive self-efficacy. Results partially support the expectation that adolescents with higher contraceptive self-efficacy act accordingly by using contraceptives. PMID- 12751311 TI - Linking early social risks to impaired physical health during the transition to adulthood. AB - The present longitudinal study of 485 youth used structural equation models to investigate the ways in which a combination of social disadvantage in the family of origin and adolescent maladjustment increases risk for physical health difficulties during adulthood. The study examined a theoretical model that proposes that disruptions in the transition to adulthood mediate the effect of earlier social disadvantage and adolescent maladjustment on young adult physical health status. Results show that early risk factors initiate a sequence of negative influences on young adult physical health through early entry into family responsibility, truncated educational attainment, and poor occupational and economic status. These associations prevailed even after controlling for physical health status during adolescence. PMID- 12751312 TI - How physician-employees experience their work lives in a changing HMO. AB - The present study advances our understanding of both physician adaptation and the physician-organization relationship in a managed care environment defined by structural diversity and constant change. It does so through a longitudinal examination of a single group of physician-employees experiencing their work lives within a nonprofit health maintenance organization (HMO) in the midst of major strategic developments. Using interview, observation, and archival data collected over a five-year period, the analysis reveals that the form and substance of individual physician adaptation to organizational life is dependent upon social exchanges over time with the HMO, making it an emergent, evolutionary process rather than a pre-determined, static phenomenon. However, the results also demonstrate that physician adaptive response to rapid, unpredictable organizational change is slow and delayed, in particular when this change makes physicians more dependent on their employing organization. This confers an advantage onto the organization vis-a-vis professionals in interpreting and responding to environmental change. These insights should encourage sociologists to employ research designs and contingency models of physician attitudes and behavior that capture the dynamic and particularistic nature of everyday physician work life in contemporary health care. PMID- 12751313 TI - Managerial and professional beliefs influencing public health privatization: results of a national survey of local health department directors. AB - This article describes managerial and professional beliefs underlying decisions to privatize public health services. We drew a stratified, nationally representative sample of local health departments and interviewed 347 department directors by telephone. We used logistic regression to establish the independent effects of various beliefs on the decision to privatize. Over half of directors did not believe that there was valid evidence that privatization results in more efficient performance, and those who believed there was such evidence were not more likely to privatize. However, directors held professional and managerial beliefs that influenced their decision to privatize. Directors most likely to privatize were those who believed that local health departments should exclusively focus on the core public health functions, those who asserted that public health should become involved in an increasingly diverse array of social problems, and those who believed that employees should be used on a temporary and contractual, rather than permanent, basis wherever possible. PMID- 12751314 TI - Phage types of Salmonella enteritidis isolated from clinical and food samples, and from broiler carcasses in southern Brazil. AB - 272 isolates of Salmonella Enteritidis (111 isolated from frozen broiler chicken carcasses, 126 from human food and other biological materials involved in food poisoning outbreaks and 35 from different poultry materials) were selected for phage typing. From these, 111 were phage typed, 57.65% being classified as phage type 4, 32.43% as phage type 4a, 3.60% as phage type 6a and 0.90% as phage type 7, whereas 5.40% samples were not phage typeable. The predominance of phage type 4 is in agreement with the results published worldwide, and reinforces the need for studies related to the epidemiological meaning of these findings. PMID- 12751315 TI - Plasmid profile in oral Fusobacterium nucleatum from humans and Cebus apella monkeys. AB - Fusobacterium nucleatum is a strict anaerobe and is indigenous of the human oral cavity. This organism is commonly recovered from different monomicrobial and mixed infections in humans and animals. In this study, the plasmid profile, the plasmid stability and the penicillin-resistance association in oral F. nucleatum isolated from periodontal patients, healthy subjects and Cebus apella monkeys were evaluated. Forty-five F. nucleatum strains from patients, 38 from healthy subjects and seven from C. apella were identified and analyzed. Plasmid extraction was performed in all the isolated strains. These elements were found in 26.7% strains from patients and one strain from C. apella. Strains from healthy subjects did not show any plasmid. Most of strains showed two plasmid bands ranging from 4 to 16 Kb, but digestions with endonucleases showed that they belonged to a single plasmid. The plasmid profile was similar and stable in human and monkey strains. Also, plasmids were classified into three groups according to size. Two strains were positive to beta-lactamase production and no plasmid DNA hybridization with a beta-lactamase gene probe was observed, suggesting a chromosomal resistance. PMID- 12751316 TI - Molecular characterization of dengue viruses type 1 and 2 isolated from a concurrent human infection. AB - In 2001, an autochthonous case of dual viremia, resulting from naturally acquired dengue virus DEN-1 and DEN-2 infections was detected during the dengue outbreak that occurred in Barretos, a city with about 105,000 inhabitants in the North region of Sao Paulo State. Serotype identification was based on virus isolation to C6/36 mosquito cells culture and immunofluorescence assays using type-specific monoclonal antibodies. The double infection was also confirmed by reverse transcriptase polymerase chain reaction (RT-PCR). Comparative analysis of the 240 nucleotide sequences of E/NS1 gene junction region between the genome of DEN-1 and DEN-2 isolates of the corresponding reference Nauru and PR 159S1 strains, respectively, showed some nucleotide differences, mainly silent mutations in the third codon position. Results of maximum likelihood phylogenetic analysis of E/NS1 gene sequences indicated that both genotypes of DEN-1 and DEN-2 viruses recovered from double infection in Barretos belonged to genotypes I and III, respectively. PMID- 12751317 TI - Molecular typing of dengue virus type 2 in Brazil. AB - Strain typing is a critical tool for molecular epidemiological analysis and can provide important information about the spread of dengue viruses. Here, we performed a molecular characterization of DEN-2 viruses isolated in Brazil during 1990-2000 from geographically and temporally distinct areas in order to investigate the genetic distribution of this serotype circulating in the country. Restriction site-specific polymerase chain reaction (RSS)-PCR presented the same pattern for all 52 Brazilian samples, showing the circulation of just one DEN-2 variant. Phylogenetic analysis using progressive pairwise alignments from 240 nucleotide sequences of the E/NS1 junction in 15 isolates showed that they belong to genotype III (Jamaica genotype). PMID- 12751318 TI - Evaluation of surrogate markers for human immunodeficiency virus infection among blood donors at the blood bank of "Hospital Universitario Regional Norte do Parana", Londrina, PR, Brazil. AB - This study evaluated the usefulness of the anti-HBc, hepatitis C virus antibodies (anti-HCV), human T cell lymphotropic virus I and II antibodies (anti-HTLV I/II), serologic tests for syphilis, and surface antigen of hepatitis B virus (HBsAg) as surrogate markers for the risk for HIV infection in 80,284 serum samples from blood donors from the Blood Bank of "Hospital Universitario Regional Norte do Parana", Londrina, Parana State, Brazil, analyzed from July 1994 to April 2001. Among 39 blood donors with positive serology for HIV, 12 (30.8%) were anti-HBc positive, 10 (25.6%) for anti-HCV, 1 (2.6%) for anti-HTLV I/I, 1 (2.6%) was positive for syphilis, and 1 (2.6%) for HBsAg. Among the donors with negative serology for HIV, these markers were detected in 8,407 (10.5%), 441 (0.5%), 189 (0.2%), 464 (0.6%), and 473 (0.6%) samples, respectively. The difference was statistically significant (p < 0.001) for anti-HBc and anti-HCV. Although the predictive positive values for these surrogate markers were low for HIV infection, the results confirmed the anti-HBc and anti-HCV as useful surrogate markers for HIV infection thus reinforcing the maintenance of them in the screening for blood donors contributing to the prevention of the small number of cases in which HIV is still transmitted by transfusion. PMID- 12751319 TI - Clinical and laboratory findings of Plasmodium vivax malaria in Colombia, 2001. AB - A descriptive study was carried out in 104 patients with Plasmodium vivax malaria, from the region of Turbo (Antioquia, Colombia). Clinical features and levels of hemoglobin, glycemia, serum bilirubin, alanine-aminotransferase (ALT), aspartate-aminotransferase (AST), creatinine and complete blood cell profile were established. 65% of the studied individuals were men and their mean age was 23. Of all individuals 59% had lived in the region for > 1 year and 91% were resident in the rural area. 42% were farmers and 35% had a history of malaria. The mean parasitaemia was 5865 parasites/mm3. The evolution of the disease was short (average of 4.0 days). Fever, headache and chills were observed simultaneously in 91% of the cases while the most frequent signs were palmar pallor (46%), jaundice (15%), hepatomegaly (17%), and spleen enlargement (12%). Anemia was found in 39% of the women and in 51% of the men, 8% of individuals had thrombocytopaenia and 41% had hypoglycemia. PMID- 12751320 TI - Accidental tetanus: prognosis evaluation in a historical series at a hospital in Salvador, Bahia, Brazil. AB - A total of 868 (84.89%) patients diagnosed with tetanus were studied, out of the 1,024 tetanus patients hospitalized at Couto Maia Hospital (Salvador, Bahia, Brazil), during the period between 1986 and 1997. Of this group (n = 868), 63.5% (n = 551) were discharged, 35.4% (n = 307) died, and 1.1% (n = 10) were transferred. The average age of the deceased patients (38.73 +/- 23.31 years) was significantly greater (p < 0.0001) than the age of those who survived (29.21 +/- 20.05 years). Analyzing the variables of the logistic regression model with statistic significance (p < or = 0.25) for univariate analysis, we observed a greater association of risk for worst prognosis (death) in patients aged > or = 51 years; time of illness < 48 hours; time of incubation < 168 hours; neck rigidity; spasms; opisthotonos; body temperature > or = 37.7 degrees C; heart beat > or = 111 beats/minute; sympathetic hyperactivity and association with pneumonia. Among the group of those who survived, patients with 1 to 5 of those variables (n = 398; 76.8%) were more frequent, while among patients of the group of the deceased, 70.3% (n = 206) presented 6 to 10 of those variables, with a highly significant difference (p < 10(-8)). In conclusion, the indicators described provide early information that may guide the prognosis and medical and nurse care. PMID- 12751321 TI - Microwave treatment of human milk to prevent transmission of Chagas disease. AB - It is recognized that breast feeding is an alternative means of transmission of Chagas disease. However, thermal treatment of milk can prevent this occurrence. As domestic microwave ovens are becoming commonplace, the efficacy of microwave thermal treatment in inactivating Trypanosoma cruzi trypomastigotes in human milk was tested. Human milk samples infected with T. cruzi trypomastigotes (Y strain) from laboratory-infected mice, were heated to 63 degrees C in a domestic microwave oven (2,450 MHz, 700 W). Microscopical and serological examinations demonstrated that none of the animals inoculated orally or intraperitoneally with infected milk which had been treated, got the infection, while those inoculated with untreated, infected milk, became infected. It was concluded that the simple treatment prescribed, which can easily be done at home, was effective in inactivating T. cruzi trypomastigotes contained in human milk. PMID- 12751322 TI - Staining of intestinal protozoa with Heidenhain's iron hematoxylin. AB - Due to its unique properties, iron hematoxylin has been traditionally recommended for staining intestinal protozoa. This process can be simplified by reducing the number of steps and periods of permanence of the slides in some of the liquids used, without detriment to the quality of the results. Thus iron hematoxylin becomes adequate for routine use. Hematoxylin is a natural dye extracted from Haematoxylon campechianum, of the family Leguminosae. It must first be 'ripened', i.e. oxidized to hematein, which reacts with ferric ammonium sulphate to produce the ferric lake (iron hematoxylin), a basic dye. Iron hematoxylin most frequently stains regressively, i.e. the slides are first overstained and then differentiated. PMID- 12751323 TI - Acute renal failure after massive honeybee stings. AB - Two clinical cases of patients who survived after numerous attacks of Africanized bees (600 and 1500 bee stings, respectively) are reported. Clinical manifestation was characterized by diffuse and widespread edema, a burning sensation in the skin, headache, weakness, dizziness, generalized paresthesia, somnolence and hypotension. Acute renal failure developed and was attributed to hypotension, intravascular hemolysis, myoglobinuria due to rhabdomyolysis and probably to direct toxic effect of the massive quantity of injected venom. They were treated with antihistaminic, corticosteroids and fluid infusion. One of them had severe acute renal failure and dialysis was required. No clinical complication was observed during hospital stay and complete renal function recovery was observed in both patients. In conclusion, acute renal failure after bee stings is probably due to pigment nephropathy associated with hypovolemia. Early recognition of this syndrome is crucial to the successful management of these patients. PMID- 12751325 TI - P system antigenic determiners expression in Ascaris lumbricoides. AB - The P System antigens have been detected in numerous parasites, bacteria and viruses, nevertheless the clinical significance is still unknown. The aim was to study the presence of P1 antigenic determiners in A. lumbricoides extracts by means of the use of 6 different monoclonal antibodies of well-known concentrations and Ig class. We worked with 14 A. lumbricoides extracts. Inhibition Agglutination Test was made in a bromelin enzymatic medium and 4 degrees C temperature. Titre, Score and Sensitivity Parameter were determined for each monoclonal antibody against red cells suspension used as revealing system. Ten extracts inhibited the agglutination of all anti P1 monoclonal antibodies. The 4 remaining extracts only inhibited the agglutination of some of them. It is demonstrated that the extracts have P1 activity. This activity is independent of titre, Score, Sensitivity Parameter, concentration and Ig class and it depends on the epitope at which the monoclonal antibody is directed. PMID- 12751324 TI - Occurrence of influenza B/Hong Kong-like strains in Brazil, during 2002. AB - Through the influenza virus surveillance from January to October 2002, influenza B/Hong Kong-like strains circulating in the Southeast and Centre East regions of Brazil have been demonstrated. This strain is a variant from B/Victoria/02/88 whose since 1991 and until recently have been isolated relatively infrequently and have been limited to South-Eastern Asia. A total of 510 respiratory secretions were collected from patients 0 to 60 years of age, with acute respiratory illness, living in the Southeast and Centre East regions of Brazil, of which 86 (17.13%) were positive for influenza virus. Among them 12 (13.95%) were characterized as B/Hong Kong/330/2001; 3 (3.49%) as B/Hong Kong/1351/2002 a variant from B/Hong Kong/330/2001; 1 (1.16%) as B/Sichuan/379/99; 1 (1.16%) as B/Shizuoka/5/2001, until now. The percentages of cases notified during the surveillance period were 34.88%, 15.12%, 15.12%, 4.65%, 15.12%, 13.95%, in the age groups of 0-4, 5-10, 11-15, 16-20, 21-30, 31-50, respectively. The highest proportion of isolates was observed among children younger than 4 years but serious morbidity and mortality has not been observed among people older than 65 years, although B influenza virus component for vaccination campaign 2002 was B/Sichuan/379/99 strain. This was probably due to the elderly protection acquired against B/Victoria/02/88. In addition, in influenza A/Panama/2007/99-like (H3N2) strains 22 (25.58%) were also detected, but influenza A(H1N1) has not been detected yet. PMID- 12751326 TI - Anticardiolipin antibodies and severe leptospirosis. PMID- 12751327 TI - Analysis of 148 kb of genomic DNA of Tetraodon nigroviridis covering an amylase gene family. AB - We have sequenced and analysed a 148 kb genomic region of Tetraodon nigroviridis, a teleost fish with a compact genome. Several genes were identified by comparison with genomic or transcript sequences of other species, informatic prediction and screening of a cDNA library. As expected for a compact genome, sizes of the identified genes and introns are very small, and intergenic distances are short. Among identified genes, three code for amylases. As in mammals, these genes are linked, but they are found in a small region of less than 11 kb. These results represent the first description of a genomic sequence larger than 100 kb in this species. Synteny with the human genome is restricted to three regions corresponding to human 1p32.3, 1p13.3 and 1p21.1. PMID- 12751328 TI - Identification of a novel apolipoprotein(a)-related protein from the European hedgehog (Erinaceus europaeus). AB - We have isolated a cDNA encoding an apo(a)-related protein designated HaRP-1 (Hedgehog apo(a) related protein-1). The HaRP-1 cDNA (2114 bp; corresponding to a 2.6 kb transcript) was isolated from a hedgehog liver cDNA library. The HaRP-1 clone corresponded to an open reading frame of 676 amino acids and contains a signal sequence followed by a preactivation domain and 7 kringle domains which exhibit an average of 57% amino acid identity with hedgehog plasminogen kringle III. We expressed HaRP-1 in human embryonic kidney cells; immunoprecipitation of metabolically-labeled conditioned medium from transfected cells showed the presence of a 74 kDa band corresponding to HaRP-1. Of note, we also observed an approximately 72 kDa species present in hedgehog plasma by western blotting using a human anti-apo(a) monoclonal antibody; we speculate that the 72 kDa plasma species corresponds to HaRP-1. Interestingly, although none of the 7 kringle domains contained a canonical lysine-binding site, we found that recombinant HaRP 1 bound specifically to lysine-Sepharose. It is likely that the evolution of the HaRP-1 gene is coincident with the evolution of hedgehog apo(a), both of which occurred by duplication of the plasminogen kringle III motif. The function of HaRP-1 remains unclear at present, but may constititute a member of the family of apo(a) proteins that functions in the regulation of lysine-dependent proteolysis. PMID- 12751329 TI - Cloning of a Twist orthologue from Enchytraeus coronatus (Annelida, Oligochaeta). AB - Enchytraeus coronatus is a small soil living oligochaete that can be maintained in culture with ease. Embryos are laid into cocoons, where they develop directly into a hatching worm within about two weeks. E. coronatus shows a simple morphology. Its transparency allows the microscopic analysis of developmental processes. To facilitate future studies on the development of specific tissues, like the mesoderm, we established molecular techniques and resources, like cDNA libraries, to allow the cloning of genes that are potentially relevant during development of the oligochaete. In this paper, we present first results using these new resources and describe the cloning of the Twist orthologue from E. coronatus. The Enchytraeus Twist protein (EcTwist) harbors all characteristic sequence features common for a true Twist orthologue. We believe that the resources described herein will facilitate phylogenetic studies on the molecular level, which will help to understand lophotrochozoan evolution. PMID- 12751330 TI - Latent periodicity of 21 bases typical for MCP II gene is widely present in various bacterial genes. AB - The existence of a typical latent periodicity of 21 bases from the Tar chemoreceptor gene of Escherichia coli (E. coli) (MCP II) in the bacterial genes has been investigated in this work. Among 583 annotated bacterial genes and ORFs in the GenBank, in which the typical periodicity has been found, the chemoreceptors' genes constituted the most numerous group (18.5%). This typical latent periodicity of 21 bases has been revealed in many different genes of regulatory proteins, DNA polymerases, reductases, kinases and others. The numbers in such gene groups varied from 1 to 4% of the total analyzed genes. The 2D structures analysis of the amino acid residues, which have been translated from the genes' regions with 21 bases periodicity, has shown that, though the enrichment of alpha-helical structures in such sequences is kept in all cases, it is seen that the latent periodicity of 21 bases is a very sensitively tuned basis, allowing the translated residues to smoothly change from one conformation to another. Interesting results have been obtained for 16S rRNAs genes of proteobacteria. Short sequences-determinants have been revealed in the genes, which select beta and gamma proteobacteria with an accuracy of above 90%. PMID- 12751332 TI - Sequencing and radiation hybrid mapping of canine uromodulin. AB - Our interest is in understanding the genetic bases for hereditary renal diseases of the domestic dog (Canis familiaris) and in characterizing gene loci for placement on the map of the canine genome. We report here on the cloning, sequencing and radiation hybrid mapping of the canine cDNA encoding uromodulin, a renal-specific glycoprotein. The cDNA is 2.3 kb in length and, as expected, comparisons of nucleotide sequences reveal that canine umod is quite similar to umod of other mammals. The predicted amino acid sequence of canine uromodulin has at least 70% identity with other mammalian uromodulin proteins. Canine umod has been mapped on the RHDF5000 radiation hybrid panel and positioned on the most recent canine genome map. Data indicate that umod is linked to the marker CZP2 (canine zona pellucida gene) on an RH group not yet assigned to a canine chromosome. The human umod and CZP2 genes are located on chromosome 16p13. PMID- 12751331 TI - Isolation, sequencing, and characterization of the cytochrome bo operon from Vitreoscilla. AB - The entire operon encoding the sodium pumping cytochrome bo from the bacterium Vitreoscilla was isolated and sequenced, and this sequence was analyzed by blast and hydropathy plots. There are fairly similar phylogenetic relationships which apply to all five proteins, but overall greater similarity to members of the gamma subdivision than the beta subdivision of the Proteobacteria. Hydropathy plots of all five Cyo proteins show near identity with those of the corresponding E. coli subunits, indicating that the similarity extends from sequence to structure. The operon appears to have a typical Shine-Dalgarno sequence, an E. coli-like promoter, and several possible binding sites for regulatory proteins. The Vitreoscilla Cyo B subunit (the probable Na+ pump) is almost identical to E. coli Cyo B at 18 key amino acids; thus, there are no obvious changes in Vitreoscilla Cyo B that hint at the details of its Na+ pumping ability. PMID- 12751333 TI - Cloning and sequencing a HemK-family gene in Porphyromonas gingivalis. AB - HemK, a universally conserved protein of unknown function, has high amino acid similarity with DNA-(adenine-N6) methyltransferases (MTases). In the present study, we sequenced a 5026 bp DNA fragment just downstream of the PgPepO gene reported previously. The DNA sequence analysis revealed three ORFs. The ORF2 gene encoded a protein of 294 amino acids with a calculated molecular weight of 32,160 Da. The deduced amino acid sequence of the ORF2 gene exhibited a significant similarity to sequence of HemK from E. coli (35% identical residues). The ORF2 gene complemented an E. coli hemK mutant. Thus, ORF2 was named PgHemK. From the point of veiw of our recent finding, that E. coli HemK catalyses the methylation of polypeptide chain release factors such as RF1 and RF2, we postulated that PgHemK might function as a protein MTase containing the DNA MTase motif. PMID- 12751334 TI - Cloning and prokaryotic expression of a cDNA encoding a putative mitochondrial malate dehydrogenase in Oryza sativa. AB - Malate dehydrogenase (MDH) has been characterized as a key player in oxaloacetate (OAA) biosynthesis mechanism in citrate acid cycle that generates reducing powers for further assimilation in the whole cell. Here we present the cloning, characterization and prokaryotic expression of a putative Mdh (OsmMDH) in Oryza sativa. Sequence alignment shows that there is a high homology between the deduced amino acid sequence of OsmMDH and MDH portein in Eucalyptus gunnii (80%), as well as between the deduced amino acid sequence of OsmMDH and other MDHs. Moreover, pI and the mitochondrial location of OsmMDH are predicted. The tissue specific expression pattern of OsmMDH reveals that it is abundant in young panicle and immature seed, while its expression level is mush lower in leaf and root. Its expression in E. coli BL21 as a fusion gene is studied further. PMID- 12751335 TI - After the hype, HOPE (and HPS): some lessons from the Women's Health Initiative trial, the Heart Outcomes Prevention Evaluation study and the Heart Prevention Study. PMID- 12751336 TI - Growth and energy and protein intake of preterm newborns in the first year of gestation-corrected age. AB - CONTEXT: There are few longitudinal studies that analyze the growth and nutritional status parameters of children born prematurely. OBJECTIVE: To evaluate the growth and dietary intake of preterm newborns in the first year of gestation-corrected age. DESIGN: Prospective clinical study. SETTING: Tertiary care hospital. PATIENTS: 19 children (7 male) who were born prematurely, with birth weight between 1000 g and 2000 g, which was adequate for the gestational age. PROCEDURES: At 3, 6, 9 and 12 months of gestation-corrected age, children were evaluated in relation to weight, height and cephalic perimeter, using the National Center for Health Statistics as the standard reference, and the Rozalez Lopez and Frisancho standards for brachial perimeter and triceps and subscapular skinfolds. The calculated dietary intake was compared to the Recommended Dietary Allowances. MAIN MEASUREMENTS: The Z score was calculated for the weight/age, height/age and weight/height relationships, and the percentiles of the perimeters and skinfolds were considered. Dietary intake records were made using the 24-hour Dietary Recall and the Food Frequency Intake Questionnaire methods. The Virtual Nutri software was used to calculate energy and protein intake. RESULTS: The weight/age, height/age and weight/height relationships and the brachial perimeter and triceps skinfold were statistically greater in the first semester in relation to the second. The cephalic perimeter remained above the 50th percentile for the ages studied and there was no difference in the subscapular skinfold between the first and second semesters, remaining below the 50th percentile. The calorie and protein intake, although statistically lower in the first than in the second semester, always remained above the recommended. CONCLUSIONS: The pace of growth is greater in the first semester than in the second, not reaching the standard expected for full-term newborns, with the exception of the cephalic perimeter, which remains adequate. Calorie/protein intake shows an inverse relationship with growth speed, remaining above the recommended for full-term newborns, although with difficulty in depositing subcutaneous fat, in spite of the high caloric intake. PMID- 12751337 TI - Effects of three different types of exercise on blood leukocyte count during and following exercise. AB - CONTEXT: High-intensity exercise causes tissue damage, production of stress hormones, and alterations in the function and quantity of various immune cells. Many clinical-physical stressors such as surgery, trauma, burns and sepsis induce a pattern of hormonal and immunological response similar to that of exercise. It has thus been suggested that heavy exercise might be used to cause graded and well-defined amounts of muscle trauma, thereby serving as an experimental model for inflammation and sepsis. OBJECTIVE: In order to explore whether some form of strenuous exercise might provide an useful model for the inflammatory process, we studied the effects of three different exercise protocols on blood leukocyte count during and following exercise. DESIGN: Four different experimental conditions, using a randomized-block design. SETTING: Defence and Civil Institute of Environmental Medicine, North York, Ontario, Canada. PARTICIPANTS: Eight healthy and moderately fit males. PROCEDURES: Participants were each assigned to four experimental conditions. Subjects performed 5 minutes of cycle-ergometry exercise at 90%, 2 hours of cycle-ergometry exercise at 60%, a standard circuit of resistance exercises with 3 sets of 10 repetitions at 60 to 70% of one repetition maximum (1-RM) force at each of 5 different stations; or they remained seated for 5 hours. DIAGNOSTIC TEST USED: Flow cytometric analysis. MAIN MEASUREMENTS: Blood samples were analyzed for total leukocyte counts, total T cells, T helper/inducer cells, T suppressor/cytotoxic cells, B cells, cytolytic T cells, and natural killer cells. RESULTS: The peak aerobic and prolonged submaximal exercise induced similar alterations in cell counts. These changes were generally larger than those produced by the resistance exercise, although both resistance and peak aerobic exercise resulted in a significantly longer lasting decrease in the CD4+/CD8+ ratio than the submaximal exercise bout did. CONCLUSION: The data suggest that, of the three exercise patterns tested, prolonged aerobic exercise induced the largest and most readily measured patterns of immune response. Nevertheless, the changes provided only a partial model for the clinical inflammatory process. PMID- 12751338 TI - Therapeutic efficacy of ranitidine bismuth citrate with clarithromycin for seven days in the eradication of Helicobacter pylori in Brazilian peptic ulcer patients. AB - CONTEXT: The curative treatment of peptic ulcer is made available nowadays through the eradication of the bacterium Helicobacter pylori, which is associated with it, but the best therapeutic regimen is yet to be determined. OBJECTIVE: To assess the efficacy of a therapeutic regimen with 400 mg ranitidine bismuth citrate associated with 500 mg clarithromycin given twice a day for seven days in a cohort of Brazilian patients with peptic ulcer. TYPE OF STUDY: Cross-sectional study. SETTING: Tertiary-care hospital. PATIENTS: One hundred and twenty nine outpatients, with active or healed peptic ulcers infected by Helicobacter pylori, diagnosed via endoscopy with confirmation via the urease test and histological examination, who had never undergone a regimen for the eradication of the bacterium. PROCEDURE: Administration of 400 mg ranitidine-bismuth and 500 mg clarithromycin twice a day, for seven days. MAIN MEASUREMENTS: Efficacy of the treatment, with a check on the cure done via another endoscopy eight weeks after drug administration. The eradication of the bacterium was determined via the urease test and histological examination. Patients who were negative for both were considered to be cured. RESULTS: Eight patients failed to complete the study. The eradication rate according to intention to treat was 81% (104/129) and per protocol was 86% (104/121). CONCLUSION: The bismuth ranitidine compound associated with clarithromycin used for one week was shown to be a simple, effective and well-tolerated therapeutic regimen for the eradication of Helicobacter pylori. PMID- 12751339 TI - Is automated platelet counting still a problem in thrombocytopenic blood? AB - CONTEXT: Reliable platelet counting is crucial for indicating prophylactic platelet transfusion in thrombocytopenic patients. OBJECTIVE: To evaluate the precision and accuracy of platelet counting for thrombocytopenic patients, using four different automated counters in comparison with the Brecher & Cronkite reference method recommended by the International Committee for Standardization in Hematology (ICSH). TYPE OF STUDY: Automated platelet counting assessment in thrombocytopenic patients. SETTING: Hematology Laboratory, Hospital do Servidor Publico Estadual de Sao Paulo, and the Hematology Division of Instituto Adolfo Lutz, Sao Paulo, SP, Brazil. MAIN MEASUREMENTS: Brecher & Cronkite reference method and four different automated platelet counters. PARTICIPANTS: 43 thrombocytopenic patients with platelet counts of less than 30,000/microliter. RESULTS: The ADVIA-120 (Bayer), Coulter STKS, H1 System (Technicom-Bayer) and Coulter T-890 automatic instruments presented great precision and accuracy in relation to laboratory thrombocytopenic samples obtained by diluting blood from normal donors. However, when thrombocytopenic patients were investigated, all the counters except ADVIA (which is based on volume and refraction index) showed low accuracy when compared to the Brecher & Cronkite reference method (ICSH). The ADVIA counter showed high correlation (r = 0.974). However, all counters showed flags in thrombocytopenic samples. CONCLUSION: The Brecher & Cronkite reference method should always be indicated in thrombocytopenic patients for platelet counts below 30,000 plt/microliter obtained in one dimensional counters. PMID- 12751340 TI - Relevance of micrometastases detected by reverse transcriptase-polymerase chain reaction for melanoma recurrence: systematic review and meta-analysis. AB - CONTEXT: Cutaneous melanoma presents significant morbidity and mortality. Nowadays, about 90% of them are diagnosed by clinical examination and most are localized melanomas. Sentinel node biopsy has brought about a new and interesting approach towards localized cutaneous melanoma. The meaning of micrometastases in sentinel nodes diagnosed by the reverse transcriptase-polymerase chain reaction is not well established. OBJECTIVE: To define the real value of micrometastases diagnosed by the reverse transcriptases polymerase chain reaction in relation to melanoma recurrence. METHODS: Systematic literature review and meta-analysis. The Cochrane Library, Medline, Embase and Lilacs were the databases searched. We used the following key words: sentinel node and melanoma; sentinel node and reverse transcriptase-polymerase chain reaction; melanoma and reverse transcriptase polymerase chain reaction. Cohort studies enrolling localized cutaneous melanoma patients who underwent sentinel node biopsy were selected. Sentinel node evaluations included hematoxylin and eosin, immunohistochemistry and reverse transcriptase-polymerase chain reaction. RESULTS: Out of the 1,542 studies evaluated, four were eligible. The four studies, when combined, were statistically homogeneous. The sample totaled 450 patients grouped as follows: 163 with a sentinel node negative to hematoxylin eosin and immunohistochemistry and positive to the reverse transcriptase-polymerase chain reaction; 192 with a sentinel node negative to hematoxylin eosin, immunohistochemistry and the reverse transcriptase-polymerse chain reaction and 95 patients with a sentinel node positive to hematoxylin eosin and/or immunohistochemistry. We analyzed the first two groups. The meta-analysis for the random model showed an increased effect from a positive reverse transcriptase-polymerase chain reaction on the recurrence rate. A similar result occurred in the meta-analysis for the fixed effect model. CONCLUSION: Patients with a positive reverse transcriptase-polymerase chain reaction had a greater recurrence rate than those with a negative reverse transcriptase-polymerase chain reaction. This suggests an important role for the reverse transcriptase-polymerase chain reaction in sentinel node examinations. In view of the small sample, a clinical trial could better evaluate this question. PMID- 12751342 TI - Crying as a precipitating factor for migraine and tension-type headache. AB - CONTEXT: Scarcely reported in the literature, crying seems to be an important precipitating factor for both migraine and tension-type headache in daily practice. OBJECTIVE: To evaluate the role of crying as a precipitating factor for migraine and tension-type headache. TYPE OF STUDY: Prospective evaluation. PARTICIPANTS: 163 workers or students from the Universidade Metropolitana de Santos, who presented at least one attack a month, for at least one year, of either migraine or tension-type headache. PROCEDURES: Interview by means of questionnaires and personal evaluations. Details of precipitating factors for the attacks were assessed. RESULTS: From the total group of 163 individuals, 90 (55.2%) considered crying to be a potential factor for triggering headache attacks. Of this group of 90 persons, 62 presented migraine (6 males, 56 females) and 28 presented tension-type headache (5 males, 23 females). Only stress, anxiety and menstrual periods rated higher or equal to crying as triggering factors for both types of headache. CONCLUSIONS: The physiology of crying is not well documented or understood. The act of crying seems to be an important precipitating factor for primary headaches and it should be studied further. The authors welcome comments on the matter and would like to work in collaboration with other groups interested in this subject. PMID- 12751341 TI - Beta-thalassemia intermedia in a Brazilian patient with-101 (C > T) and codon 39 (C > T) mutations. AB - CONTEXT: We verified molecular alterations in a 72-year-old Brazilian male patient with a clinical course of homozygous beta-thalassemia intermedia, who had undergone splenectomy and was surviving without regular blood transfusions. The blood cell count revealed microcytic and hypochromic anemia (hemoglobin = 6.5 g/dl, mean cell volume = 74 fl, mean cell hemoglobin = 24 pg) and hemoglobin electrophoresis showed fetal hemoglobin = 1.3%, hemoglobin A2 = 6.78% and hemoglobin A = 79.4%. OBJECTIVE: To identify mutations in a patient with the symptoms of beta-thalassemia intermedia. DESIGN: Molecular inquiry into the mutations possibly responsible for the clinical picture described. SETTING: The structural molecular biology and genetic engineering center of the Universidade Estadual de Campinas, Campinas, Brazil. PROCEDURES: DNA extraction was performed on the patient's blood samples. The polymerase chain reaction (PCR) was done using five specific primers that amplified exons and the promoter region of the beta globin gene. The samples were sequenced and then analyzed via computer programs. RESULTS: Two mutations that cause the disease were found: -101 (C > T) and codon 39 (C > T). CONCLUSIONS: This case represents the first description of 101 (C > T) mutation in a Brazilian population and it is associated with a benign clinical course. PMID- 12751343 TI - Pregnancy and delivery in a patient with aortic prosthesis for Leriche syndrome. AB - CONTEXT: Leriche syndrome is a thrombotic obliteration of the bifurcation of the aorta, a rare condition that usually affects older men as a result of atherosclerosis. Women of childbearing age rarely need a vascular prosthesis (as a result of Leriche syndrome or other conditions) and there is no literature on an association between Leriche syndrome/vascular prosthesis and pregnancy/labor/delivery. CASE REPORT: A case of pregnancy and delivery in a 38 year-old patient with Leriche syndrome and an aortoiliac prosthesis is presented. The patient had no complications during pregnancy, and was admitted to the maternity hospital when close to term, to begin heparin therapy. Labor ensued spontaneously and a normal vaginal delivery occurred, resulting in a healthy infant. The authors present their considerations regarding the delivery route and the rationale for deciding in favor of vaginal childbirth. PMID- 12751345 TI - Psoriatic arthritis: a guide for dermatology nurses. AB - Psoriatic arthritis (PsA) affects people in the prime of life, causing functional impairment and diminished quality of life. Etanercept, the first FDA-approved therapy, and other immunobiologics, offer hope for favorable long-term outcomes. It is imperative that dermatology nurses learn more about the basic immunology of psoriasis and PsA and the role of immunomodulation in their treatments. PMID- 12751346 TI - Quality of life considerations in psoriasis treatment. AB - Psoriasis can have a profound impact on a patient, interfering in all aspects of life. Therefore, measuring the impact of disease and the effects of treatment must include both physiologic measurements as well as health-related quality of life tools. Psychosocial evaluation of patients at risk allows for early interventions that will promote positive patient outcomes and compliance with the treatment pathways. PMID- 12751347 TI - Cutaneous metastasis of rectal adenocarcinoma. AB - Cutaneous metastasis of rectal adenocarcinoma is a rare event occurring in fewer than 4% of all patients with rectal cancer. When present, it typically signifies disseminated disease with a poor prognosis. Occasionally patients present with skin metastases with no evidence of visceral involvement. In these patients, the early detection and proper diagnosis of metastatic rectal cancer can significantly alter treatment and prognosis. PMID- 12751348 TI - What's your assessment? Candida balanitis. PMID- 12751349 TI - Epidermolysis bullosa: the challenges of wound care. AB - Epidermolysis bullosa (EB) is a severe blistering skin disorder. Care for an individual diagnosed with EB can be challenging. Wound care for the more severe types of EB can consume an individual's life; therefore, one of the most important nursing considerations is to educate individuals and family members about proper wound care, products, and different dressing techniques. PMID- 12751351 TI - Wound assessment and evaluation: diabetic ulcer protocol. PMID- 12751350 TI - Percutaneous absorption of topically applied medication. AB - Given the central role that topical medications play in treating skin disease, nurses must understand the delivery systems of topical preparations to make the most informed decisions for their patients and provide adequate patient education to ensure the best clinical outcomes. PMID- 12751352 TI - Dermatology nursing scope of practice. AB - The intent of this document is to conceptualize practice and provide education to practitioners, educators, researchers, and administrators, and to inform other health professionals, legislators, and the public about the participation in and contribution to health care by dermatology nursing. Through articulation of the elements of care, dimensions, boundaries, and intersection, the Dermatology Nursing Scope of Practice document defines the specialty practice of dermatology nursing. PMID- 12751353 TI - Dermatology nursing standards of clinical practice. PMID- 12751354 TI - Herpes zoster. PMID- 12751355 TI - Birt-Hogg-Dube syndrome. PMID- 12751356 TI - A rash is more than skin deep. AB - The principles of dermatologic examination focus on visual recognition, diagnosis, and treating the presenting skin problem. Psychologic dysfunction may not be immediately obvious. PMID- 12751357 TI - Pyoderma gangrenosum. PMID- 12751358 TI - The Pittsburgh Sleep Quality Index. PMID- 12751359 TI - Alien abductions as a risk factor for Parkinson's disease [April Fool]. PMID- 12751360 TI - The intimidating voice of blood. PMID- 12751361 TI - Diabetes mellitus: a 2003 perspective. PMID- 12751362 TI - Type 2 diabetes mellitus in children and adolescents: an emerging problem. PMID- 12751363 TI - Eighty years of insulin therapy: 1922-2002. PMID- 12751364 TI - Achieving glycemic control in young children with type 1 diabetes: approaches, pitfalls and new technologies. AB - Advances in technologies for insulin administrations, glucose monitoring, development of an artificial pancreas and cell-based therapy will ultimately have a profound effect on the lives of people wit diabetes. There is both current success and substantial promise, indicating that these approches may offer, for the first time, real potential for achieving euglycemia without hypoglycemia. Given the physiological and psychosocial impact of type 1 diabetes in young children, this group of patients and their parents stand to gain especially great benefit from these developments. However, the potential for improvements in the mangement of diabetes in young children based on available technologies should not be overlooked and should be effectively utilized as the standard for patient care. Only twenty years ago blood glucose reagent strips were first coming into routine use. Current meters have greatly reduced the amount of blood required ( now less than 1 microliter for many meters) and greatly imporved precision. The advent modified, recombinant insulins, which became available only in the last several years, allows for an insulin regimen to better match the absorption of dietary carbohydrate. All technologies have improved our ability to attain glycemic control, thereby reducing the risk of long-term complication in even our youngest patients. PMID- 12751365 TI - Managing diabetic dyslipidemia. PMID- 12751366 TI - Enhancing the care of diabetes in primary care. PMID- 12751367 TI - Dilated eye examinations for people with diabetes--wherein lies the truth? PMID- 12751368 TI - Images in medicne. Fournier's gangrene. PMID- 12751369 TI - Tumour-associated hypermethylation: silencing E-cadherin expression enhances invasion and metastasis. PMID- 12751370 TI - Quality assurance in radiotherapy. AB - In 1999, the European Organisation for Research and Treatment of Cancer (EORTC), being a European pioneer in the field of cancer research as well as in quality assurance (QA), launched an Emmanuel van der Schueren fellowship for QA in radiotherapy. In this paper, the work that has been done during the first E. van der Schueren fellowship is reported, focusing on four phase III EORTC clinical trials: 22921 for rectal cancer, 22961 and 22991 for prostate cancer and 22922 for breast cancer. A historical review of the QA programme of the EORTC Radiotherapy group during the past 20 years is included. PMID- 12751371 TI - Radiotherapy quality assurance: time for everyone to take it seriously. AB - Like high-risk industries, radiotherapy requires intense attention to detail, alertness, precision, and adequate human and material resources to minimise the risk of irreversible consequences. Clinical trials data such as that generated by the Quality Assurance programme of the Radiotherapy Group of the European Organization for Research and Treatment of Cancer (EORTC) in this issue of the Journal have been instrumental in identifying problems with technical quality, the understanding of which can have a direct impact on improving the quality of care in the community. Consistency in absolute dosimetry, dose delivery, volume definition and reproducibility are paramount in radiotherapy quality assurance and have become even more important with the advent of conformal therapy. Extension of these principles to other oncological disciplines has added an additional dimension of improvement. Waiting times and measures of access must also be monitored if overall quality at the population level is to be assessed and enhanced. Lessons should be learned from clinical trials methodology in the use of intervention-specific guidelines, physician education and real time audit of treatment planning decisions. In the future, novel approaches, such as web based systems may further improve education and audit. Wider application and audit of evidence-based management guidelines about the use radiotherapy will bring to standard clinical practice the quality benefits that are considered a basic minimum standard for clinical trials. PMID- 12751372 TI - The Quality Assurance programme of the Radiotherapy Group of the European Organization for Research and Treatment of Cancer (EORTC): a critical appraisal of 20 years of continuous efforts. AB - In 1982, the European Organization for Research and Treatment of Cancer (EORTC) Radiotherapy Group established the Quality Assurance (QA) programme. During the past 20 years, QA procedures have become a major part of the activities of the group. The methodology and steps of the QA programme over the past 20 years are briefly described. Problems and conclusions arising from the results of the long lasting QA programme in the EORTC radiotherapy group are discussed and emphasised. The EORTC radiotherapy group continues to lead QA in the European radiotherapy community. Future challenges and perspectives are proposed. PMID- 12751373 TI - Predictive value of tumour cell proliferation in locally advanced breast cancer treated with neoadjuvant chemotherapy. AB - We previously reported that defects in apoptotic pathways (mutations in the TP53 gene) predicted resistance to doxorubicin monotherapy. The aim of this study was to evaluate whether cell proliferation, as assessed by mitotic frequency and Ki 67 levels, may provide additional predictive information in the same tumours and to assess any potential correlations between these markers and mutations in the TP53 gene and erbB-2 overexpression. Surgical specimens were obtained from ninety locally advanced breast cancers before commencing primary chemotherapy consisting of weekly doxorubicin (14 mg/m2) for 16 weeks. 38% of the patients had a partial response (PR) to therapy, 52% had stable disease (SD) while 10% had progressive disease (PD). Univariate analysis showed a significant association between a high cell proliferation rate (expressed as a high mitotic frequency) and resistance to doxorubicin (P = 0.001). Further analyses revealed this association to be limited to the subgroup of tumour expressing wild-type TP53 (P = 0.016), and TP53 mutation status was the only factor predicting drug resistance in the multivariate analyses. The finding that a high mitotic frequency, as well as a high Ki-67 staining, correlated to TP53 mutations (P = 0.001 for both), suggests TP53 mutations are the key predictor of drug resistance, although cell proliferation may play an additional role in tumours harbouring wild-type TP53. Regarding overall (OS) and relapse-free survival (RFS), multivariate analyses (Cox' proportional hazards regression) revealed a high histological grade and negative oestrogen receptor (ER) status to be the variables that were most strongly related to breast cancer death (P = 0.001 and P = 0.001, respectively). A key reason for this difference with respect to the factors predicting chemotherapy resistance could be due to the adjuvant use of tamoxifen in all patients harbouring ER-positive tumours. PMID- 12751374 TI - A mutant TP53 gene status is associated with a poor prognosis and anthracycline resistance in breast cancer patients. AB - This study evaluates the prognostic and predictive relevance of a mutated p53 in a series of 254 samples from primary breast cancer patients. C-erbB-2 analysis was defined in a limited subpopulation of 79 patients. p53 and c-erbB-2 status was analysed by immunohistochemical staining of the tumour samples. Positive p53 immunostaining was present in 86 cases (34%) and correlated with a high malignant grade, negative progesterone receptor status and ductal histology of tumour. C erbB-2 positivity was seen in 38 samples (48%). Within an average follow-up time of 74 months, 121 patients developed recurrent or metastatic disease. Patients with mutated p53 showed a statistically significant shorter overall survival and disease-free survival in both univariate and multivariate analyses. The worst clinical outcome was seen in patients who were both p53- and c-erbB-2-positive. The response rate to anthracycline-based chemotherapy in metastatic disease was low in the p53-positive cases. Our results help to clarify the independent prognostic role of a mutated p53 status in breast cancer patients, indicating that this gene might be predictive of anthracycline resistance. Patients with a mutant p53 status and overexpressing c-erbB-2 should be regarded as high-risk cases. PMID- 12751375 TI - Elevated plasma levels of transforming growth factor-beta 1 (TGF-beta 1) in patients with advanced breast cancer: association with disease progression. AB - We examined the association between an elevated plasma TGF-beta 1 level and the disease progression of advanced breast cancer (BC) patients (n = 44). TGF-beta 1 levels were detected by an enzyme-linked immunosorbent assay (ELISA). Platelet carryover and in vitro platelet activation in our plasma samples was assessed and found to be insignificant. Plasma TGF-beta 1 values were significantly elevated (P < 0.05) in stage IIIB/IV patients (median value: 2.40 ng/ml, range: 0.13-8.48 ng/ml, n = 44) compared with healthy donors (median value: 1.30 ng/ml, range: 0.41-4.93 ng/ml, n = 36). Although pronounced in metastatic patients, especially those who had been newly diagnosed, TGF-beta 1 elevation was independent of tumour mass, site of distant metastases, histopathological type, steroid receptor (SR) content and age of the BC patients. Follow-up of 6 patients indicated a relationship between the plasma TGF-beta 1 and the patient's response. This suggests that TGF-beta 1, may be a promising prognostic marker for breast cancer patients with advanced disease. Confirmatory large-scale studies are needed, particularly given the overlap of values between our different subgroups analysed. PMID- 12751376 TI - Pathological features of breast cancer response following neoadjuvant treatment with either letrozole or tamoxifen. AB - Morphological characteristics, grading features, proliferation marker MIB1, apoptosis (by Tdt-mediated duTP-biotin nick-end labelling (TUNEL)), Bcl-2 expression, oestrogen receptor (ER) and progesterone receptor (PgR) status were compared in ER-positive breast cancers before and after 3 months of neoadjuvant therapy with either letrozole or tamoxifen. Daily treatment was with letrozole 2.5 mg (12 patients) or 10 mg (12 patients), or with tamoxifen 20 mg(24 patients). Letrozole treatment was associated with a pathological response in 17 of 24 (71%) patients. The predominant change in grading features was a decrease in mitosis, and the expression of MIB1 was reduced in all of the 22 evaluable cases. Whilst only marginal changes were observed in ER expression following letrozole therapy, PgR reactivity was reduced in 20 of 21 evaluable cases which were initially PgR-positive, becoming undetectable in 16 patients. Tamoxifen treatment was associated with pathological response in 15 of 24 (63%) tumours. In contrast to letrozole, the dominant change in grading feature was an increase in tubule formation, ER score was markedly reduced in most cases, and the most common effect on PgR was an increased expression. Following treatment with either tamoxifen or letrozole, variable effects were observed on the apoptotic index and expression of Bcl-2. These results indicate that both letrozole and tamoxifen have marked influences on the pathological features of breast cancer during neoadjuvant therapy. However, the effects of the two agents varied such that the phenotypes of letrozole- and tamoxifen-treated tumours differ markedly. Effects on clinical, pathological and biological endpoints were frequently disconcordant- future studies will therefore require the evaluation of multiple parameters in order to fully assess tumour response. PMID- 12751377 TI - CD103+ intraepithelial lymphocytes--a unique population in microsatellite unstable sporadic colorectal cancer. AB - Colorectal cancers with microsatellite instability (MSI) typically show increased numbers of intraepithelial lymphocytes (IEL) in comparison to microsatellite stable (MSS) cancers. The aim of this study was to determine the phenotype of this unique lymphocyte population in MSI and MSS colorectal cancers. Twenty-four individuals with sporadic colorectal cancer (17 MSI, 7 MSS) were included in this study. Intraepithelial and stromal lymphocytes were detected using immunohistochemistry with anti-CD8 and anti-CD103 antibodies, and two observers independently quantified the numbers of lymphocytes. CD103+ (alpha E beta 7+) IELs detected within tumour tissue co-expressed CD8+ while the stromal lymphocytes were phenotypically heterogeneous, with respect to CD8+ and CD103+ expression. MSI colorectal cancers harboured increased numbers of CD8+ CD103+ IELs, as well as CD8+ CD103- and CD8+ CD103+ stromal lymphocytes, when compared with MSS colorectal cancers. CD103+ IELs were found at 27-fold greater numbers in the tumour epithelium than in normal epithelium from the same patient (P = 0.001, Wilcoxon matched pairs test). From our findings, we have proposed a mechanism for the homing of these alpha E beta 7+ lymphocytes to tumour tissue in MSI and MSS colorectal cancers. PMID- 12751378 TI - Brain metastases following interleukin-2 plus interferon-alpha-2a therapy: a follow-up study in 94 stage IV melanoma patients. AB - This study analyses the frequency and therapy of brain metastases in 94 stage IV melanoma patients after treatment with high-dose interleukin-2 (IL-2) and interferon-alpha (IFN-alpha) within three subsequent trials between 1990 and 1995. Central nervous system (CNS) metastases occurred in 28 patients (30%) during the potential follow-up period of 6 years. Time to occurrence of brain metastases varied between 1 and 53 months, with a median of 10 months. Of 28 patients, 19 had < 5 metastases, which were treated with stereotactic radiosurgery (SR) in 9 patients. In 2 patients, SR was followed by resection. 9 patients had multiple metastases, of which 4 received whole brain irradiation (WBI). Median survival after the detection of CNS metastases was 6 months (95% Confidence Interval (CI) 1-11 months). SR plus resection was associated with a prolonged survival of 34 and 35 months in 2 patients, 1 patient survived for 41 months after WBI, demonstrating the efficacy of these therapeutic strategies. PMID- 12751379 TI - Cisplatin, doxorubicin and ifosfamide in carcinosarcoma of the female genital tract. A phase II study of the European Organization for Research and Treatment of Cancer Gynaecological Cancer Group (EORTC 55923). AB - Carcinosarcomas of the female genital tract are highly malignant tumours composed of carcinomatous and sarcomatous elements. In the past, these tumours were frequently treated as sarcomas. However, a number of arguments, including the sensitivity of these tumours to platinum-based chemotherapy, suggest that these tumours behave more like poorly differentiated carcinomas. The European Organization for Research and Treatment of Cancer (EORTC) Gynaecological Cancer Group therefore decided to perform a prospective phase II study in patients with advanced or metastatic carcinosarcoma with an approach such as that used in gynaecological carcinomas. Eligible patients could have primary or recurrent disease, but prior radiotherapy or chemotherapy was not allowed. The treatment plan recommended upfront debulking, followed by chemotherapy with cisplatin, ifosfamide and doxorubicin. Patients who could be debulked to non-measurable disease remained eligible for the study, but the response assessment was restricted to patients who had measurable disease before the start of chemotherapy. A total of 48 patients (39 primary disease, 9 recurrent disease) were registered, 41 of them being eligible. In 9 patients, all macroscopic lesions could be removed, 32 patients were left with residual disease and were assessable for response. The overall response rate was 56%: a complete response (CR) was observed in 11 (34%) patients and partial response (PR) in 7 (22%) patients. No change occurred in 5 patients and progression in 2 patients. In 7 patients, response could not be assessed. Median survival for all of the 41 eligible patients was 26 months. Severe leucopenia and thrombocytopenia were common and necessitated dose reductions or delays in 60% of patients. From a clinical point of view, the most severe non-haematological toxicity was renal dysfunction, and one patient died of this complication in the absence of disease progression. The results of this study are in-line with the hypothesis that carcinosarcomas are chemosensitive, in particular for the currently investigated regimen. The treatment also included upfront cytoreduction when feasible. Considering the observed toxicities, alternative platinum-based regimens with more favourable toxicity profiles should be explored. PMID- 12751380 TI - Scandinavian Sarcoma Group Osteosarcoma Study SSG VIII: prognostic factors for outcome and the role of replacement salvage chemotherapy for poor histological responders. AB - From 1990 to 1997, 113 eligible patients with classical osteosarcoma received neo adjuvant chemotherapy consisting of high-dose methotrexate, cisplatin and doxorubicin. Good histological responders continued to receive the same therapy postoperatively, while poor responders received salvage therapy with an etoposide/ifosfamide combination. With a median follow-up of 83 months, the projected metastasis-free and overall survival rates at 5 years are 63 and 74%, respectively. Independent favourable prognostic factors for outcome were tumour volume < 190 ml, 24-h serum methotrexate > 4.5 microM and female gender. The etoposide/ifosfamide replacement combination did not improve outcome in the poor histological responders. In conclusion, this intensive multi-agent chemotherapy results in > 70% of patients with classical osteosarcoma surviving for 5 years. The data obtained from this non-randomised study do not support discontinuation and exchange of all drugs used preoperatively in histological poor responders. As observed in previous Scandinavian osteosarcoma studies, female gender appears to be a strong predictor of a favourable outcome. PMID- 12751381 TI - Trends in tobacco smoking among adolescents in Lyon, France. AB - To provide information that may promote more effective cancer prevention, we identified factors associated with regular smoking among adolescents in Lyon, France. School grades where these factors began to influence regular smoking were also identified. Seven consecutive cross-sectional anonymous surveys were conducted in three public schools, beginning in grade 6eme (average age 11.5 years) in 1993 and ending in grade Terminale (average age 17.4 years) in 1999. All classes in each respective grade were surveyed, with 3650 completed questionnaires for all years combined. Prevalence of current regular smoking is presented according to school grade for 17 variables identified as significantly related to regular smoking in a multivariate logistic regression analysis. Important factors associated with regular smoking were identified as early as grade 6eme and included not viewing the taking care of one's health as important, not eating breakfast regularly, associating with groups where smoking occurs, having a best friend who smokes, and having a brother and/or sister who smokes. Not regularly reading was first associated with an increased risk of regular smoking in grade 5eme. Not living with both parents, alcohol drinking, episodes of drunkenness, illicit drug use, and sexual relationships were positively associated with regular smoking in the middle and later grades, when these questions were first asked. Not playing sports and not playing with computers were initially associated with an increased risk of regular smoking in grade 3eme. PMID- 12751382 TI - Family history of cancer and risk of ovarian cancer. AB - The aim of this study was to examine the relationship between history of cancer in first-degree relatives and ovarian cancer risk. Between 1992 and 1999, we conducted a case-control study in Italy on 1031 women with epithelial ovarian cancer and 2411 women admitted to hospital for acute non-neoplastic conditions. Odds ratios (OR) were estimated using unconditional logistic regression, adjusted for age and several potential confounders. Overall, 27 cases and nine controls reported a family history of ovarian cancer (OR = 7.0; 95% confidence interval (CI) 3.1-16). The OR was 23 (95% CI 2.6-212) below age 50 years, based on 10 cases and one control only. The risk of ovarian cancer was also increased in women with a family history of cancer of the stomach (OR = 1.5; 95% CI 1.0-2.1), intestine (OR = 1.7; 95% CI 1.2-2.4), lung (OR = 1.3; 95% CI 1.0-1.8), breast (OR = 2.3; 95% CI 1.7-3.1), lymphomas (OR = 2.3; 95% CI 1.0-5.1) and all sites (OR = 1.6; 95% CI 1.4-1.9). Our results confirm the higher ovarian cancer risk in women with a family history of ovarian and breast cancer, and suggest a few associations with other sites. PMID- 12751383 TI - Viral infection, atopy and mycosis fungoides: a European multicentre case-control study. AB - Mycosis fungoides (MF) is a rare disease with an unknown aetiology, although it has been suggested that infections may play a role. The present study investigates whether infections, atopic disorders and some other diseases are risk indicators for MF. A European multicentre case-control study involving seven rare cancers, including MF, was conducted from 1995 to 1998. Patients between 35 and 69 years of age diagnosed with MF (n = 140) were recruited, and the diagnoses were verified by a reference pathologist, who classified 83 cases as definitive and 35 cases as possible; 22 cases were not accepted. Of the 118 accepted cases, 104 patients were interviewed (including 76 definitive cases and 28 possible cases). These 76 definitive cases were used for this study. A common set of controls to serve all case groups were interviewed, representing a total of 4574 controls. The latter included 1008 colon cancer patients and 3566 subjects selected from population registers. Information on infections, skin pathology and clinical history 5 years before the diagnosis of MF was used to estimate odds ratios (ORs) derived from logistic regression-modelling, which included gender, age and country. The highest ORs for MF were found in patients who reported a history of psoriasis 5 years before MF was diagnosed (OR 7.2, 95% CI: 3.6-14.5). Urticaria had an OR of 1.4 (95% CI: 0.6-3.6). Infections and atopic diseases were not closely associated with MF. Some diseases correlated to MF. Whether this has a causal background or reflects early diagnostic uncertainty is not known. PMID- 12751385 TI - The association of E-cadherin expression and the methylation status of the E cadherin gene in nasopharyngeal carcinoma cells. AB - Loss of E-cadherin (E-cad) has been associated with progression and poor survival in nasopharyngeal carcinoma (NPC). In this study, we investigated the role of methylation on E-cad inactivation in NPC cell lines, as well as in NPC tissue samples. Using 6 NPC cell lines, we found that methylation of the E-cad 5' CpG island promoter region was correlated with the loss of both mRNA and E-cad protein expression in these cell lines. In addition, using 29 NPC and 10 non malignant nasopharyngeal samples, we also observed 5' CpG methylation of the E cad gene in 52% (15 out of 29) NPC samples, but in only 10% (1 out of 10) of the non-malignant nasopharyngeal tissues. Our findings indicate that 5' CpG island methylation of the E-cad gene may play an important part in the inactivation of E cad in NPC. Our results also suggest that reducing the methylation of the E-cad gene may be a potential therapeutic strategy for NPC. PMID- 12751384 TI - E-cadherin expression is silenced by DNA methylation in cervical cancer cell lines and tumours. AB - A previous study showed E-cadherin expression was lost in some cervical cancer cell lines and tumours. This study was designed to clarify the significance of DNA methylation in silencing E-cadherin expression. We examined promoter methylation of E-cadherin in five cervical cancer cell lines and 20 cervical cancer tissues using methylation-specific PCR (MSP) and bisulphite DNA sequencing. The correlation of E-cadherin methylation and expression together with methyltransferase (DNMT1) were further studied. We found that hypermethylation of E-cadherin was involved in five cervical cancer cell lines and 40% (8/20) of cervical cancer tissues. E-cadherin protein was lost in 6/8 (75%) samples and 3/5 (60%) cell lines with promoter methylation. E-cadherin methylation was significantly correlated with increased DNMT1. Using an antisense DNMT1 oligo to transfect into SiHa HeLa C33A cell line, E-cadherin protein was re expressed. We concluded that loss of E-cadherin expression was in part correlated with DNA methylation and DNMT1 expression in cervical cancer. PMID- 12751386 TI - Increased tumour extracellular pH induced by Bafilomycin A1 inhibits tumour growth and mitosis in vivo and alters 5-fluorouracil pharmacokinetics. AB - The aim was to determine if a specific inhibitor of vacuolar H(+)-ATPases (V ATPases), Bafilomycin A1 (BFM), could increase the low extracellular pH (pHe) typical of solid tumours and thus inhibit their growth in vivo. BFM inhibited the proliferation of various human cells and rat pituitary GH3 tumour cells in vitro (IC50: 2.5-19.2 nM), and flow cytometry on GH3 cells showed a marked increase in S and G2M phases after 16-48 h, but no evidence of increased apoptosis. BFM caused significant inhibition of GH3 xenograft growth, and histomorphometry showed a 30% decrease in mitosis but no change in apoptosis. 31P-magnetic resonance spectroscopy (MRS) in vivo of GH3 xenografts showed that BFM increased pHe, but did not affect pHi, resulting in a decrease in the negative pH gradient (-delta pH). BFM decreased lactate formation suggesting a reduction in glycolysis. We suggest that BFM reduces extracellular H(+)-transport by inhibition of V-ATPases leading to an increase in pHe and decreased glycolysis, and thus reduced tumour cell proliferation. 19F-MRS in vivo showed that a smaller -delta pH was associated with decreased retention of 5-fluorouracil (5FU) which was consistent with our previous data in vivo implying the -delta pH controls tumour retention of 5 FU. PMID- 12751387 TI - Significance of dihydropyrimidine dehydrogenase activity in renal cell carcinoma. AB - Dihydropyrimidine dehydrogenase (DPD) is the rate-limiting enzyme in the pathway of uracil and thymine catabolism. DPD is also the principal enzyme involved in the degradation of 5-fluorouracil (5-FU), an anticancer chemotherapeutic agent that is used clinically to treat renal cell carcinoma (RCC). Little is known about the significance of DPD activity in human cancers. We investigated the activity of DPD in 68 RCC and the relationship between DPD activity and the sensitivity to 5-FU. The levels of DPD activity in RCC and normal kidney samples were determined by the 5-FU degradation assay. The sensitivity to 5-FU was assessed by the microculture tetrazolium dye assay. The activity of DPD was approximately 2-fold higher in normal kidney compared with RCC. DPD activity in Stage I/II RCC was approximately 2-fold higher than that in Stage III/IV RCC. The levels of DPD activity in Grade 1 and Grade 2 RCC were 3 and 2-fold higher, respectively, than that in the Grade 3 cancers. There was an inverse correlation between DPD activity in RCC cells and their sensitivity to 5-FU. Furthermore, 5 chloro-2,4-dihydroxypyridine (CDHP), a potent DPD inhibitor, enhanced the sensitivity to 5-FU. The current study is the first to demonstrate that the level of DPD activity was inversel correlated with both the progression of the disease and increased grade of RCC, and that DPD activity was inversely associated with the sensitivity of RCC to 5-FU, which was enhanced by a DPD inhibitor. These results suggest that a low DPD activity may be associated with the malignant potential of RCC. In addition, it may be possible to overcome 5-FU resistance by using DPD inhibitors in the treatment protocols of 5-FU-based chemotherapy for RCC. PMID- 12751388 TI - A comment and update on "Does the histological subtype of high-grade central osteosarcoma influence the response to treatment with chemotherapy and does it affect overall survival?". PMID- 12751389 TI - An analysis of the risk of B-lymphocyte malignancies in industrial cohorts. AB - Among numerous studies of occupational groups with varied chemical exposures (e.g., farmers, petroleum workers, and rubber workers), some have reported excess risk for non-Hodgkin's lymphoma (NHL), multiple myeloma, and other cancers of the B-lymphocyte cell line. While not conclusive, these studies raise questions about the effects of chemical exposures on the lymphocytic versus myeloid cell lines. Almost 70 occupational cohort studies were identified that addressed B-cell cancer risks in 9 major industrial categories, in order to look for common patterns across industries. This effort was substantially limited by the inconsistent nature of lymphohematopoietic (LH) classification schemes across studies and over time, and the relative paucity of B-cell-specific results in studies for any given industry. Taking these limitations into consideration, a descriptive, graphical analysis suggested a pattern of B-cell cancer elevations in the rubber and "general chemical" industries, but no consistent patterns in petroleum production/distribution or petrochemical production. The limited data sources, which lack detail about differences in hazard and exposure for different types of products/chemicals, did not allow a comprehensive look at possible common exposures associated with B-cell cancer elevations across industries. This study suggests that evaluation of possible associations between specific chemical exposures and B-cell malignancies would require additional studies with clear and common definitions of B-cell outcomes. The article concludes by giving an example of a possible common framework for categorizing NHL, the diseases for which most classification issues arise. PMID- 12751391 TI - More. PMID- 12751390 TI - CYP1A induction and blue sac disease in early developmental stages of rainbow trout (Oncorhynchus Mykiss) exposed to retene. AB - Early life stages of rainbow trout were exposed to different regimes of water borne retene (7-isopropyl-1-methylphenanthrene) to determine if there is an ontogenic stage particularly sensitive to retene toxicity, and if cytochrome P 4501A (CYP1A) induction is a forerunner to blue sac disease (BSD), the syndrome of toxicity. CYP1A protein concentrations, measured by immunohistochemistry, were first detected during organogenesis, when organ and enzyme systems are first being developed, and steadily increased until swim-up. The prevalence of signs of BSD rose 1 wk following a marked increase in CYP1A activity after hatch, suggesting that CYP1A induction is related to BSD. The larval stage was the most sensitive to retene toxicity, based on CYP1A induction and a high prevalence of BSD. The most common signs of BSD were hemorrhaging, yolk-sac edema, and mortality, but hemorrhaging was the first and most frequently observed response. Tissue concentrations of retene were elevated just after fertilization, but decreased steadily as fish developed to the swim-up stage, most likely due to the establishment of more efficient metabolic and excretory systems in later stages of development. PMID- 12751392 TI - Family presence: evidence versus tradition. PMID- 12751393 TI - Early predictors of long-term disability after injury. AB - BACKGROUND: Improving outcomes after serious injury is important to patients, patients' families, and healthcare providers. Identifying early risk factors for long-term disability after injury will help critical care providers recognize patients at risk. OBJECTIVES: To identify early predictors of long-term disability after injury and to ascertain if age, level of disability before injury, posttraumatic psychological distress, and social network factors during hospitalization and recovery significantly contribute to long-term disability after injury. METHODS: A prospective, correlational design was used. Injury specific information on 63 patients with serious, non-central nervous system injury was obtained from medical records; all other data were obtained from interviews (3 per patient) during a 2 1/2-year period. A model was developed to test the theoretical propositions of the disabling process. Predictors of long term disability were evaluated using path analysis in the context of structural equation modeling. RESULTS: Injuries were predominately due to motor vehicle crashes (37%) or violent assaults (21%). Mean Injury Severity Score was 13.46, and mean length of stay was 12 days. With structural equation modeling, 36% of the variance in long-term disability was explained by predictors present at the time of injury (age, disability before injury), during hospitalization (psychological distress), or soon after discharge (psychological distress, short term disability after injury). CONCLUSIONS: Disability after injury is due partly to an interplay between physical and psychological factors that can be identified soon after injury. By identifying these early predictors, patients at risk for suboptimal outcomes can be detected. PMID- 12751394 TI - Naturalistic decision making: a model to overcome methodological challenges in the study of critical care nurses' decision making about patients' hemodynamic status. AB - The quality of critical care nurses' decision making about patients' hemodynamic status in the immediate period after cardiac surgery is important for the patients' well-being and, at times, survival. The way nurses respond to hemodynamic cues varies according to the nurses' skills, experiences, and knowledge. Variability in decisions is also associated with the inherent complexity of hemodynamic monitoring. Previous methodological approaches to the study of hemodynamic assessment and treatment decisions have ignored the important interplay between nurses, the task, and the environment in which these decisions are made. The advantages of naturalistic decision making as a framework for studying the manner in which nurses make decisions are presented. PMID- 12751395 TI - Endotracheal suctioning with or without instillation of isotonic sodium chloride solution in critically ill children. AB - BACKGROUND: Instillation of isotonic sodium chloride solution for endotracheal tube suctioning is controversial. Research has focused on the effect of such instillation in adults; no studies in children have been published. OBJECTIVES: (1) To describe differences in oxygen saturation depending on whether or not isotonic sodium chloride solution is instilled during suctioning and (2) to describe the rates of occlusion of endotracheal tubes and nosocomial pneumonia. METHODS: A convenience sample of 24 critically ill patients were enrolled before having suctioning and after informed consent had been given. Ages ranged from 10 weeks to 14 years. Patients were randomized to 1 of 2 groups. In group 1, subjects received between 0.5 and 2.0 mL of isotonic sodium chloride solution, depending on their age, once per suctioning episode. In group 2, subjects received no such solution. A total of 104 suctioning episodes were analyzed. Oxygen saturation was recorded at predetermined intervals before and for 10 minutes after suctioning. Occlusion of endotracheal tubes and rates of nosocomial pneumonia also were compared. RESULTS: Patients who had isotonic sodium chloride solution instilled experienced significantly greater oxygen desaturation 1 and 2 minutes after suctioning than did patients who did not. No occlusions of endotracheal tubes and no cases of nosocomial pneumonia occurred in either group. CONCLUSIONS: Results of this study support a growing body of evidence that instillation of isotonic sodium chloride solution during endotracheal tube suctioning may not be beneficial and actually may be harmful. PMID- 12751396 TI - A multisite survey of suctioning techniques and airway management practices. AB - BACKGROUND: Ventilator-associated pneumonia, common in critically ill patients, is associated with microaspiration of oropharyngeal secretions and may be related to suctioning and airway management practices. OBJECTIVES: To describe institutional policies and procedures related to closed-system suctioning and airway management of intubated patients, and to compare practices of registered nurses and respiratory therapists. METHODS: A descriptive, comparative, multisite study of facilities that use closed-system suctioning devices on most intubated adults was conducted. Nurses and respiratory therapists who worked at the sites completed surveys related to their practices. RESULTS: A total of 1665 nurses and respiratory therapists at 27 sites throughout the United States responded. The typical respondent had at least 6 years' experience with patients receiving mechanical ventilation (61%) and a baccalaureate degree or higher (54%). Most sites had policies for management of endotracheal tube cuffs (93%), hyperoxygenation (89%) and use of gloves (70%) with closed-system suctioning, and instillation of isotonic sodium chloride solution for thick secretions (74%). Only 48% of policies addressed oral care and 37% addressed oral suctioning. Nurses did more oral suctioning and oral care than respiratory therapists did, and respiratory therapists instilled sodium chloride solution more and rinsed the suctioning device more often than nurses did. CONCLUSIONS: Policies vary widely and do not always reflect current research. Consistent performance of practices such as wearing gloves for airway management and maintaining endotracheal cuff pressures must be evaluated. Collaborative, research-based policies and procedures must be developed and implemented to ensure best practices for intubated patients. PMID- 12751397 TI - Legionnaires disease: a case study. PMID- 12751398 TI - Do-not-resuscitate and stratification-of-care forms in Rhode Island. AB - Congress passed the Patient Self-Determination Act of 1990 to ensure that patients are informed of their rights to express healthcare preferences in advance of loss of capacity. Thus, a patient may elect to forgo cardiopulmonary resuscitation in favor of a treatment approach that favors comfort over survival. Do-not-resuscitate and stratification-of-care forms provide a means for expression of healthcare preferences in hospitals. These forms can often guide the important discussion of healthcare preferences. Unfortunately, no clear standard exists for what should be included in do-not-resuscitate or stratification-of-care forms that institutions seeking to improve in this vital area of practice could use for guidance. Existing forms in use at adult general hospitals throughout Rhode Island were reviewed. PMID- 12751399 TI - Small-bowel arteriovenous shunting due to systemic lupus erythematosus vasculitis. PMID- 12751400 TI - Family presence during cardiopulmonary resuscitation and invasive procedures: practices of critical care and emergency nurses. AB - BACKGROUND: Increasingly, patients' families are remaining with them during cardiopulmonary resuscitation and invasive procedures, but this practice remains controversial and little is known about the practices of critical care and emergency nurses related to family presence. OBJECTIVE: To identify the policies, preferences, and practices of critical care and emergency nurses for having patients' families present during resuscitation and invasive procedures. METHODS: A 30-item survey was mailed to a random sample of 1500 members of the American Association of Critical-Care Nurses and 1500 members of the Emergency Nurses Association. RESULTS: Among the 984 respondents, 5% worked on units with written policies allowing family presence during both resuscitation and invasive procedures and 45% and 51%, respectively, worked on units that allowed it without written policies during resuscitation or during invasive procedures. Some respondents preferred written policies allowing family presence (37% for resuscitation, 35% for invasive procedures), whereas others preferred unwritten policies allowing it (39% for resuscitation, 41% for invasive procedures), Many respondents had taken family members to the bedside (36% for resuscitation, 44% for invasive procedure) or would do so in the future (21% for resuscitation, 18% for invasive procedures), and family members often asked to be present (31% for resuscitation, 61% for invasive procedures). CONCLUSIONS: Nearly all respondents have no written policies for family presence yet most have done (or would do) it, prefer it be allowed, and are confronted with requests from family members to be present. Written policies or guidelines for family presence during resuscitation and invasive procedures are recommended. PMID- 12751401 TI - Creating a more responsible public dialogue about the social, ethical, and legal aspects of genomics. PMID- 12751402 TI - A quick test predicts acute coronary events. AB - Inflammation of the coronary arterial wall plays a major role in atherosclerosis and ultimately thrombosis by contributing to vascular constriction, spasm, and thrombus formation. Measurement of hs-CRP level is a readily available laboratory blood test that serves as a gauge of coronary plaque inflammation. As a result, hs-CRP has become a very useful biological marker for predicting the risk of acute coronary events and for making decisions regarding treatment. It is important to recognize that lipid-lowering therapy decreases plaque inflammation and slows the progression of calcium buildup in the coronary arteries, which is readily verified by reduction in hs-CRP levels. Endothelial dysfunction is a product of plaque inflammation and as such can predict acute CV events. Endothelial function can be assessed during cardiac catheterization by measuring the vasoactive response to pharmacological or physiological stress. However, the routine use of cardiac catheterization to measure drug-induced coronary vasoactivity can have potential adverse effects in patients with unstable coronary disease. Time and costs can also be additional constraints in the routine use of this procedure. Consequently the simple and readily available hs CRP test is accurate and preferable. PMID- 12751403 TI - Interpreting a postoperative 12-lead ECG waveform. PMID- 12751404 TI - JCAHO to conduct unannounced resurveys in all accreditation programs in 2006. PMID- 12751405 TI - Results on early 2003 surveying of National Patient Safety Goals. PMID- 12751406 TI - Standards Review Project nears completion. PMID- 12751407 TI - Top compliance issues for all of 2002. PMID- 12751408 TI - Regulations passed by the board and/or amended during 2002. PMID- 12751410 TI - Earn CME credit for outpatient quality improvement projects. PMID- 12751412 TI - Severe Acute Respiratory Syndrome (SARS) in China and status of scientific and clinical knowledge, 14 April 2003. PMID- 12751413 TI - Human plague in 2000 and 2001. PMID- 12751414 TI - Micronutrient deficiency--an underlying cause of morbidity and mortality. PMID- 12751415 TI - Contribution of breastfeeding to vitamin A nutrition of infants: a simulation model. AB - OBJECTIVE: To provide information on the potential contribution to vitamin A nutrition in infants of strategies for improving maternal vitamin A status and increasing the consumption of breast milk. METHODS: The contribution of breastfeeding to the vitamin A nutrition of children in eight age groups between 0 and 24 months was simulated under four sets of conditions involving two levels of breast milk consumption with or without maternal vitamin A supplementation. FINDINGS: During the first 6 months, optimal breastfeeding on its own (compared with withholding colostrum and then partially breastfeeding after the first week) was as effective as postpartum maternal supplementation alone, retinol intakes being increased by 59 micrograms per day and 68 micrograms per day, respectively. Combined in synergy, these strategies increase retinol intake by 144 micrograms per day, or 36% of the recommended intake. After 6 months, partial breastfeeding continued to provide a significant proportion of the recommended intakes: 42% from 6-12 months and 61% during the second year. CONCLUSION: Maternal supplementation with a high dose of vitamin A at the time of delivery and the promotion of optimal breastfeeding practices are highly effective strategies for improving vitamin A nutrition in infants and should be strengthened as key components of comprehensive child survival programmes. PMID- 12751416 TI - Community-based monitoring of safe motherhood in the United Republic of Tanzania. AB - OBJECTIVE: To examine the progress made towards the Safe Motherhood Initiative goals in three areas of the United Republic of Tanzania during the 1990s. METHODS: Maternal mortality in the United Republic of Tanzania was monitored by sentinel demographic surveillance of more than 77,000 women of reproductive age, and by prospective monitoring of mortality in the following locations; an urban site; a wealthier rural district; and a poor rural district. The observation period for the rural districts was 1992-99 and 1993-99 for the urban site. FINDINGS: During the period of observation, the proportion of deaths of women of reproductive age (15-49 years) due to maternal causes (PMDF) compared with all causes was between 0.063 and 0.095. Maternal mortality ratios (MMRatios) were 591 1099 and maternal mortality rates (MMRates; maternal deaths per 100,000 women aged 15-49 years) were 43.1-123.0. MMRatios in surveillance areas were substantially higher than estimates from official, facility-based statistics. In all areas, the MMRates in 1999 were substantially lower than at the start of surveillance (1992 for rural districts, 1993 for the urban area), although trends during the period were statistically significant at the 90% level only in the urban site. At the community level, an additional year of education for household heads was associated with a 62% lower maternal death rate, after controlling for community-level variables such as the proportion of home births and occupational class. CONCLUSION: Educational level was a major predictor of declining MMRates. Even though rates may be decreasing, they remained high in the study areas. The use of sentinel registration areas may be a cost-effective and accurate way for developing countries to monitor mortality indicators and causes, including for maternal mortality. PMID- 12751417 TI - Decentralization and equity of resource allocation: evidence from Colombia and Chile. AB - OBJECTIVE: To investigate the relation between decentralization and equity of resource allocation in Colombia and Chile. METHODS: The "decision space" approach and analysis of expenditures and utilization rates were used to provide a comparative analysis of decentralization of the health systems of Colombia and Chile. FINDINGS: Evidence from Colombia and Chile suggests that decentralization, under certain conditions and with some specific policy mechanisms, can improve equity of resource allocation. In these countries, equitable levels of per capita financial allocations at the municipal level were achieved through different forms of decentralization--the use of allocation formulae, adequate local funding choices and horizontal equity funds. Findings on equity of utilization of services were less consistent, but they did show that increased levels of funding were associated with increased utilization. This suggests that improved equity of funding over time might reduce inequities of service utilization. CONCLUSION: Decentralization can contribute to, or at least maintain, equitable allocation of health resources among municipalities of different incomes. PMID- 12751418 TI - Household willingness to pay for azithromycin treatment for trachoma control in the United Republic of Tanzania. AB - OBJECTIVE: Household willingness to pay for treatment provides important information for programme planning. We tested for relationships between socioeconomic status, risk of trachoma, perceptions of the effects of azithromycin, and the household willingness to pay for future mass treatment with azithromycin. METHODS: We surveyed 394 households in 6 villages located in central United Republic of Tanzania regarding their willingness to pay for future azithromycin treatment. A random sample of households with children under 8 years of age was selected and interviewed following an initial treatment programme in each village. Data were gathered on risk factors for trachoma, socioeconomic status, and the perceived effect of the initial azithromycin treatment. Ordered probit regression analysis was used to test for statistically significant relationships. FINDINGS: 38% of responding households stated that they would not be willing to pay anything for future azithromycin treatment, although they would be willing to participate in the treatment. A proxy for cash availability was positively associated with household willingness to pay for future antibiotic treatment. Cattle ownership (a risk factor) and being a household headed by a female not in a polygamous marriage (lower socioeconomic status) were associated with a lower willingness to pay for future treatment. A perceived benefit from the initial treatment was marginally associated with a willingness to pay a higher amount. CONCLUSIONS: As those at greatest risk of active trachoma indicated the lowest willingness to pay, imposing a cost recovery fee for azithromycin treatment would likely reduce coverage and could prevent control of the disease at the community level. PMID- 12751420 TI - The epidemics of bovine spongiform encephalopathy and variant Creutzfeldt-Jakob disease: current status and future prospects. AB - The large epidemic of bovine spongiform encephalopathy (BSE) in the United Kingdom has been in decline since 1992, but has spread to other countries. The extensive control measures that have been put in place across the European Union and also in Switzerland should have brought the transmission of BSE under control in these countries, provided that the measures were properly enforced. Postmortem tests on brain tissue enable infected animals to be detected during the late stages of the incubation period, but tests that can be performed on live animals (including humans) and that will detect infections early are urgently needed. The number of infected animals currently entering the food chain is probably small, and the controls placed on bovine tissues in the European Union and Switzerland should ensure that any risks to human health are small and diminishing. Vigilance is required in all countries, especially in those in which there has been within species recycling of ruminant feed. Fewer than 150 people, globally, have been diagnosed with variant Creutzfeldt-Jakob disease (vCJD), but there are many uncertainties about the future course of the epidemic because of the long and variable incubation period. Better control measures are necessary to guard against the possibility of iatrogenic transmission through blood transfusion or contaminated surgical instruments. These measures will required sensitive and specific, diagnostic tests and improved decontamination methods. PMID- 12751419 TI - Quality and comparison of antenatal care in public and private providers in the United Republic of Tanzania. AB - OBJECTIVE: To compare the quality of public and private first-tier antenatal care services in Dar es Salaam, United Republic of Tanzania, using defined criteria. METHODS: Structural attributes of quality were assessed through a checklist, and process attributes, including interpersonal and technical aspects, through observation and exit interviews. A total of 16 health care providers, and 166 women in the public and 188 in the private sector, were selected by systematic random sampling for inclusion in the study. Quality was measured against national standards, and an overall score calculated for the different aspects to permit comparison. FINDINGS: The results showed that both public and private providers were reasonably good with regard to the structural and interpersonal aspects of quality of care. However, both were poor when it came to technical aspects of quality. For example, guidelines for dispensing prophylactic drugs against anaemia or malaria were not respected, and diagnostic examinations for the assessment of gestation, anaemia, malaria or urine infection were frequently not performed. In all aspects, private providers were significantly better than public ones. CONCLUSION: Approaches to improving quality of care should emerge progressively as a result of regular quality assessments. Changes should be introduced using an incremental approach addressing few improvements at a time, while ensuring participation in, and ownership of, every aspect of the strategy by health personnel, health planners and managers and also the community. PMID- 12751421 TI - Shifting the burden: the private sector's response to the AIDS epidemic in Africa. AB - As the economic burden of human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) increases in sub-Saharan Africa, allocation of the burden among levels and sectors of society is changing. The private sector has more scope to avoid the economic burden of AIDS than governments, households, or nongovernmental organizations, and the burden is being systematically shifted away from the private sector. Common practices that transfer the burden to households and government include pre-employment screening, reductions in employee benefits, restructured employment contracts, outsourcing of low skilled jobs, selective retrenchments, and changes in production technologies. Between 1997 and 1999 more than two-thirds of large South African employers reduced the level of health care benefits or increased employee contributions. Most firms also have replaced defined-benefit retirement funds, which expose the firm to large annual costs but provide long-term support for families, with defined contribution funds, which eliminate risks to the firm but provide little for families of younger workers who die of AIDS. Contracting out previously permanent jobs is also shielding firms from benefit and turnover costs, effectively shifting the responsibility to care for affected workers and their families to households, nongovernmental organizations, and the government. Many of these changes are responses to globalization that would have occurred in the absence of AIDS, but they are devastating for the households of employees with HIV/AIDS. We argue that the shift in the economic burden of AIDS is a predictable response by business to which a deliberate public policy response is needed. Countries should make explicit decisions about each sector's responsibilities if a socially desirable allocation is to be achieved. PMID- 12751422 TI - Frameworks for studying the determinants of child survival. PMID- 12751423 TI - How private practitioners help to control TB. PMID- 12751424 TI - Deadlock on access to cheap drugs at global trade negotiations. PMID- 12751425 TI - Donors are distorting India's health priorities, say protestors. PMID- 12751426 TI - Leprosy elimination in India inches closer. PMID- 12751427 TI - Antiretroviral misuse in Mumbai, India. PMID- 12751428 TI - Task complexity and limb selection in reaching. AB - Research examining limb selection for reaching and grasping an object in various positions of hemispace has noted a strong ipsilateral bias for using the hand on the same side as the stimulus, an observation that to some extent questions the traditional notion of handedness. The present study examined the effects of task complexity in regard to such actions. Forty-four right-handed, blindfolded subjects were required to grasp a small cube at one location and release it at another site, with movement initiated only after the second cue was presented. In condition A, the first tone identified the location where the subject was to grasp the cube (the second tone, the release point), while in condition B the order was reversed. A view of both conditions revealed a similar trend. As expected, the vast majority (average 96%) used their dominant (right) hand to reach into right hemispace and release in the left field. However, when the reach cue was presented in left hemispace, only 40% of subjects selected their nondominant (ipsilateral) limb to complete the action. Since this value is substantially less than reported previously with a less complex task (70%), we speculate that when deeper processing is required there is a tendency to revert to the dominant limb, even when it necesssitates reaching contralaterally. Additional discussion focuses on programming selection for reach and release. PMID- 12751429 TI - The protective mechanisms of the basic rest-activity cycle as an indirect manifestation of this rhythm in waking: preliminary report. AB - The hypothesis of the Basic Rest Activity Cycle (BRAC), that underlies approximately 90-min fluctuations in states of consciousness, has gained strong documentation during sleep in cyclical occurrence of its REM and NON-REM stages, however, there is no convicting evidence of this rhythm in waking. According to the concept of protective mechanisms of BRAC (Kokoszka, 1990), the rest phase of the BRAC manifests in states of consciousness with domination of passive state of mind and spontaneous activity of imagination, as well as domination contemplation over goal oriented activity, that occur during some socially accepted everyday activities. In order to verify this, 30 persons (21 women and 9 men) aged between 19-52 (M = 29.1; SD = 10.08) answered specially designed sets of questions three times a day (at noon, at 5 p.m., and before sleep) over 2 consecutive days. The results indicate that states considered as mechanisms of information metabolism were reported by all subjects and with a mean frequency of 9.0 (SD = 1.5) on the first day, and 8 (SD = 1.65) on the second day. Most often they were classified as purposeful overstimulation, thoughtlessness, and fatigue. PMID- 12751430 TI - A new extra-vertebral treatment model for incomplete spinal cord injuries. AB - Advances made in recent times in spinal cord injury repair research will soon take us toward a cure in paraplegics. But what are the prospects for quadriplegics? Certain fundamental issues make treatment approaches to quadriplegia different and difficult. Injury at cervical region poses additional problems for any surgical intervention with life-threatening risks of i) endangering respiratory function, ii) cavitation, cysts, and syringomyelia formation extending cephalad to the injury, and iii) mid-lower cervical injuries, lower motor neuron death, and the resultant degeneration of brachial plexus axons would still leave the upper limbs denervated and paralyzed even as treatment procedures might successfully salvage the lower limbs. With these apparently insurmountable impediments in quadriplegic cord repair, it would be wise to turn to alternative treatment strategies. Conventional treatment models since the days of Ralph Gerard (1940) have all used intra-vertebral procedures. We present here a plausible extra-vertebral repair model suitable for incomplete cord injuries at cervical, thoracic, and lumbar levels. The procedure consists of identifying the extent of viable grey-white matter in the injured area and to utilize it efficiently as a "neural tissue bridge." Next, labile state is induced by using botulinum toxin/colchicine (Krishnan, 1983, 1991; Krishnan et al., 2001 a,b) and Ca+ channel blockers in the motorsensory nerve terminals of polisegmentally innervated skeletal muscles that "bridge" the injured cord segments. This would retrogradely induce a redundant state of intra-spinal growth of nerve terminals and new synaptic connections within those viable neural tissues, as well as promote effective relinking of the injured cord ends and enhance motor-sensory recovery. PMID- 12751431 TI - Synaptic strengthening and continuum activity-wave growth in temporal sequencing during cognitive tasks. AB - Sequencing, the distinction between "before" and "after," is a basic feature of cognition, and is exemplified most simply by classical or Pavlovian conditioning. It is generally accepted that such learned behavior is connected anatomically to synaptic strengthening, as first postulated by Hebb. However, the original Hebbian formulation has difficulties, most prominently the assumption that synaptic connections between individual afferent and efferent neurons while apparently modified by learning, has no mechanism that is presently understood. There is also the problem that there is, in general, a long synaptic path between the cortical areas associated with the conditioned and unconditioned reflexes. Neural continuum theory, which deals with elements containing large numbers of neurons, provides an alternative explanation, namely that synaptic strengthening occurs on a larger scale, involving the neurons in regions of consistently substantial activity. The neural continuum has the property of amplifying waves of wavelength large compared with synaptic connection ranges, which propagate over distances comparable to the physical separation between the cortical areas associated with the components of a cognitive task. In this example, propagating waves generated by the conditioned and unconditioned reflexes interfere, producing a standing wave in an intermediate geometric region concave toward the (temporally later) unconditioned reflex. Synaptic strengthening in this region has the effect of a "lens" focusing the activity generated by the conditioning stimulus toward the motor region associated with the conditioned response, producing the Pavlovian reflex. PMID- 12751432 TI - Investigation of the laterality of hostility, cardiovascular regulation, and auditory recognition. AB - This experiment tested a hypothesis linking the right cerebral regulation of hostility and cardiovascular arousal. First, replication of previous research supporting heightened cardiovascular (systolic blood pressure [SBP] and diastolic blood pressure [DBP]) reactivity among high hostile participants was partially successful. Second, dynamic variations in functional cerebral asymmetry in response to emotional linguistic processing was measured. Thirty low- and high hostile, undergraduate volunteer participants (n = 30) were identified using the Cook Medley Hostility Scale (CMHS). Only healthy, right-handed male participants completed the experiment. All participants completed the negative affective auditory verbal learning test (AAVLT). Cardiovascular measures (SBP and DBP) were recorded and dichotic listening procedures were administered before and after the cognitive affective stressor. The results support greater left cerebral activation among both groups following the dichotic phoneme listening tasks and greater right cerebral activation among both groups following an emotional linguistic (affective verbal learning) cognitive stressor. PMID- 12751433 TI - Trail making test cut-offs for malingering among cocaine, heroin, and alcohol abusers. AB - The Trail Making Test (TMT) is frequently used to screen for cognitive impairments in substance abusers; however, an existing problem is that substance abusers may give poor effort and the TMT results may not be valid. In this study, cutting scores for malingering were developed from three samples drawn from electronic files of data from the Drug Abuse Treatment Outcome Study (DATOS), a naturalistic, prospective cohort study that collected data from 1991-1993 in 96 drug abuse treatment programs in 11 cities in the United States. The DATOS enrolled 7689 substance abusers. The three drawn samples were for subjects with primary drugs of abuse. Number of subjects were as follows: alcohol-1000, cocaine/crack-4306, heroin-1548. Data were analyzed to determine number of substance abusers that fell beyond the upper end of the distribution of TMT scores at the ten, five, and one percentiles. These percentiles were set for alcoholics, cocaine abusers, and heroin abusers. The proper use of the cut-off scores is to alert clinicians to the increasingly higher probability of poor effort when a substance abuser in one of the three groups scores beyond the one percent cut-off or his or her sample of primary drug of abuse. Clearly, the use of these cut-offs needs further empirical validation. PMID- 12751434 TI - Hypothalamic digoxin, cerebral chemical dominance, and pathogenesis of pulmonary diseases. AB - The isoprenoid pathway is a key regulatory pathway in the cell. It synthesizes digoxin, an endogenous membrane Na(+)-K+ ATPase inhibitor and modulator of synaptic transmission. The role of the isoprenoid pathway in lung diseases and its relation to hemispheric dominance was assessed in this study. The following parameters were measured in patients with (i) bronchial asthma, (ii) chronic bronchitis emphysemia, (iii) idiopathic pulmonary fibrosis, (iv) sarcoidosis, and (v) in individuals with right hemispheric, left hemispheric and bihemispheric dominance: 1. plasma HMG CoA reductase, digoxin, dolichol, ubiquinone, and magnesium levels, 2. tryptophan, tyrosine catabolic patterns, 3. free radical metabolism, 4. glycoconjugate metabolism, and 5. membrane composition. In patients with lung disease there was elevated digoxin synthesis, increased dolichol and glycoconjugate levels, and low ubiquinone and elevated free radical levels. The RBC membrane Na(+)-K+ ATPase activity and serum magnesium were decreased. There was also an increase in tryptophan catabolites and reduction in tyrosine catabolites in the serum. There was an increase in cholesterol:phospholipid ratio and a reduction in glycoconjugate level of RBC membrane in these patients. The same biochemical patterns were obtained in individuals with right hemispheric chemical dominance. An upregulated isoprenoid pathway and hyperdigoxinemia are characteristic of lung disease and right hemispheric chemical dominance. Right hemispheric chemical dominance is important in deciding the predisposition to lung disease. PMID- 12751435 TI - Hypothalamic mechanisms of immunity. AB - The present article considers a synthetical analysis of the results reported by our laboratory in the last twenty years in the field of neuroimmunomodulation. The studies we discuss here continue a previous research activity, a synthesis of which has also been published in this journal (Baciu, 1988). In that paper, we reported data concerning the role of the hypothalamic tubero-mammillary area in triggering of the phagocytic and of the secondary immune specific response. Here, we present an analysis of experimental facts gathered after 1988, and also of some prior to that date, which were not included in the above-mentioned review. They regard localizations, attained with stereotactical methods, of hypothalamic areas involved in maintenance of basal phagocytosis and of its circadian rhythm, of the phagocytic and of the primary and secondary specific response. We attempted to re-analyze these data in an integrative view, and accomplish a coherent image of the hypothalamic mechanisms of the nonspecific and specific immune response. The conclusion we draw is that the nervous system may exert its modulatory action upon the immune response in several ways: i) subsequent to a direct hypothalamic stimulation (electrical or through bacteria or bacterial products) or to a cortico-hypothalamic stimulation; ii) depending on the nature, intensity, duration, and frequency of the appropriate stimulus, it may either enhance the immune response, via neural and humoral pathways, or depress it; iii) via the hypothalamus-pituitary-adrenal axis. Nervous triggering and enhancement of the immune response are essential, their occurrence in the initial stages ensuring its favorable course. The finding that repeated electroconvulsant shocks, employed for hypothalamus stimulation in dogs of different breed, age, weight, and individual history, are followed by extremely variable changes of the phagocytic activity raises the question on the individuality of the immune response. PMID- 12751436 TI - Hypothalamic digoxin, hemispheric chemical dominance, and addictive behavior. AB - The isoprenoid pathway produces an endogenous membrane sodium-potassium ATPase inhibitor, digoxin. Digoxin can regulate neurotransmitter transport in the brain with upregulation of tryptophan transport over tyrosine. The pathway was assessed in individuals with addiction, as well as in those with differing hemispheric dominance. The isoprenoid pathway was upregulated with increased digoxin synthesis in addiction. There was an increase in tryptophan catabolites- quinolinic acid, serotonin, nicotine, and strychnine--in patients with addiction, and a reduction in tyrosine catabolites--dopamine, noradrenatine, and morphine. The pattern seen in addiction individuals was similar to that in right hemispheric chemical dominance. Addiction represents a state of right hemispheric chemical dominance, hyperdigoxinemia, and endogenous morphine deficiency state. PMID- 12751437 TI - Wound bed preparation. PMID- 12751438 TI - The antimicrobial benefits of silver and the relevance of microlattice technology. PMID- 12751439 TI - The mode of action of a superabsorbent polymer wound dressing (TenderWet). PMID- 12751441 TI - Debridement rates with activated polyacrylate dressings (TenderWet). PMID- 12751440 TI - Clinical experience using silver antimicrobial dressings on venous stasis ulcers. PMID- 12751442 TI - Preparing the wound for grafting. PMID- 12751443 TI - No-care equals bad care. A talk with Sam Bozzette. Interview by Bob Huff. PMID- 12751445 TI - HAART to heart talk. PMID- 12751444 TI - HIV pathogenesis reports from the 10th Retrovirus Conference. PMID- 12751446 TI - ConFuzeon reigns. PMID- 12751447 TI - A report on the International Treatment Preparedness Summit. PMID- 12751448 TI - You have HIV...and you have AIDS. PMID- 12751449 TI - AIDS directors seek help to fight epidemic in the South. Call to action for more money, resources, research. AB - As AIDS spreads rapidly in the South, a new picture is emerging, which shows the epidemic is becoming increasingly rural, female, African-American, and poor. Two new studies are helping identifying why so many rural Southerners avoid the health care systems and don't trust their doctors about HIV infection. Clinicians need to address high rates of depression and post-traumatic stress syndrome in HIV-infected patients living in the rural South. PMID- 12751450 TI - High rates of trauma, distrust in rural HIV-positives. PMID- 12751451 TI - New testing strategy to help track HIV. Detuned testing initiated at 24 sentinel sites. AB - The CDC is moving quickly to monitor HIV incidence across the country using its detuned testing technology, with nearly $6 million this year committed to 24 sentinel cities. PMID- 12751452 TI - Syphilis role in HIV being studies in California. STD and HIV surveillance working together. AB - For the past three years, syphilis outbreaks in men who have sex with men have worried health official because of what they indicate about a resurgence in high risk behaviors. But are these outbreaks facilitating HIV transmission, or is syphilis contained mostly to MSM who are HIV-positive? PMID- 12751454 TI - High-income countries see increase in epidemic. Italy sees increase in sexual transmission. AB - The HIV/AIDS epidemic has grown to include about 1.6 million people who live in high-income countries, including the estimated 76,000 people who became infected with the virus in 2002. PMID- 12751453 TI - U.S. AIDS funding is smoke and mirrors, critics charge. Distribution could take more than a year. AB - In January, when President Bush announced at his State of the Union address that he would provide a five-year, $15 billion Emergency Plan for AIDS Relief worldwide, the efforts received international praise. However, when the FY 2004 budget proposal was released, critics were calling the Bush plan a shell game. PMID- 12751455 TI - CDC ramps up HIV behavioral surveillance. Data collection begins this summer in 15 cities. PMID- 12751456 TI - Coalition seeks funding for HCV/HIV co-infection. Integrating services makes economic, medical sense. AB - With Hepatitis C virus infecting up to 40% of HIV-positive patients, liver disease has become the leading cause of death in AIDS patients. And yet funding for hepatitis screening and treatment is inadequate, and states must use creative strategies to integrate HCV screening into existing HIV services. PMID- 12751457 TI - States going at slow pace developing hepatitis plans. AB - Hindered by lack of federal funds and patient advocacy, only a few states have hepatitis prevention plans in place, according to a recently completed survey. Despite high rates of hepatitis infection in HIV-positive patients, nearly half of surveyed states offered no hepatitis C virus counseling and testing at publicly funded sites. PMID- 12751458 TI - Resources for integrating HCV and HIV services. PMID- 12751459 TI - Computer tool helps plan cost-effective strategies. Most important variable is HIV incidence. AB - The Rand Corporation has developed a computer program to help community planning groups and health departments set priorities among the wide array of available HIV interventions. The tool's central component is an Excel spreadsheet for entering local data, such as HIV prevalence and demographic profiles. PMID- 12751460 TI - FDA news. Price criticized for new anti-HIV drug. AB - The FDA approved a new class of anti-HIV drugs that will benefit patients whose virus has become resistant to other treatments. But as states are facing funding shortfalls for AIDS drug assistance programs, activists say most patients won't be able to afford the drug. PMID- 12751461 TI - Proceedings of the 2nd International Workshop on Waldenstrom's Macroglobulinemia. Athens, Greece, September 2002. PMID- 12751463 TI - [Chemically acquired retinopathy: The committee of the American Academy of Ophthalmology re-enter the debate]. PMID- 12751462 TI - [External serology quality control programs developed in Latin America with the support of PAHO from 1997 through 2000]. AB - OBJECTIVE: To evaluate the quality of serological screening of blood donors in five groups of blood banks in Latin America that participated over the 1997-2000 period in an external serology control project developed with support from the Pan American Health Organization (PAHO). METHODS: With assistance from PAHO, the Serology Authority of the Pro-Blood Foundation/Blood Center of Sao Paulo (Fundacao Pro-Sangue/Hemocentro de Sao Paulo), of Sao Paulo, Brazil, carried out the external quality control project and served as its "organizing center" (OC). The OC developed five external serology quality control "programs" (ESQCPs), or external evaluation activities, for the respective groups of participating blood banks. There was one ESQCP each in 1997, 1999, and 2000, and there were two in 1998. In these five programs, the number of participating blood banks ranged from 13 to 21, and the number of countries ranged from 11 to 16. In each program, the OC used a set of 24 blinded sera samples with different reactivities for the various infectious agents for which screening is obligatory in Brazil. Each participating institution in each program received a sera set, to be processed using that institution's standard screening procedures. After returning its results to the OC, each participant received an answer key for the sera set, to be used in evaluating its own performance. All the individual results were kept strictly confidential. At the end of each program, the OC prepared and sent to all the participants a final report that contained information on the overall results from that program. RESULTS: An analysis of the five programs showed that there was a lack of homogeneity among the countries with respect to the strategies and the parameters used in screening blood donors. Few laboratories screened for human T-cell-lymphotropic virus (beginning with the 1997 program, the respective rates were 17%, 27%, 35%, 39%, and 45%). Rates of screening were also low for antibodies to the hepatitis B core antigen (again, beginning with the 1997 program, the rates were 42%, 27%, 39%, 50%, and 60%). There were also important differences with respect to which tests and which combinations of tests were used, making it hard to compare the types of screening done. In the five programs, with the various tests used, the overall rate of false positive results fluctuated around 2%. The highest false positive rate for any of the tests, 4.6%, was for antibodies against the hepatitis C virus. The lowest false positive rate, 0.4%, was for antibodies against Trypanosoma cruzi. CONCLUSIONS: These results show the need for PAHO to continue using these external quality control programs as well as other activities in order to strengthen the procedures for serological screening blood banks in Latin America, until there is more uniformity in the procedures that the countries use. PMID- 12751464 TI - [The transfusion medicine program in Cuba]. AB - Cuba's Transfusion Medicine Program (TMP) is a subsystem of the country's National Health System. The TMP's objective is to ensure hemotherapy with blood that is safe and sufficient for all the individuals who need it. The TMP subsystem is made up of the National Commission on Transfusion Medicine, the Institute of Hematology and Immunology, 37 clinical services, 44 blood banks, 120 collection centers, 19 mobile units, and 37 blood certification laboratories. Additional facilities include a laboratory for plasma separation, a laboratory that produces leukocyte interferon and transfer factor, and two laboratories that produce reagents for blood classification and blood diagnosis symptoms. In Cuba, blood donation is voluntary. Since 1997 approximately 5% of the population per year has donated blood, thus meeting the goal recommended by the Pan American Health Organization of one voluntary blood donation annually for every 20 persons. During 2002, 563,204 blood donations were received, and there were 445,898 transfusions of blood or blood components. All donations are individually screened for HIV 1 and 2, hepatitis B, hepatitis C, and syphilis, thus meeting the country's current regulations. In 2002 these screening measures led to discarding, respectively, 0.12%, 0.60%, 0.71%, and 1.8% of the blood donations. Although the prevalence of human T-cell lymphotropic virus I and II in Cuba is very low, this test will soon be added to the screening process. PMID- 12751465 TI - [Peru's experience with a national blood banking program]. AB - This paper describes Peru's experiences with its National Blood Banking Program. Until the mid-1990s, the country faced a host of problems, including the lack of a legal framework to regulate blood banks, a high maternal mortality rate due to a shortage of blood, virtually no voluntary donations, a high risk of infection from transfusions, the use of only whole blood for transfusion, serious disorganization in the blood banks, deficiencies in blood bank supervision and control, no training programs, indifference on the part of health officials, frequent selling of blood, and limited community awareness. Subsequently, a strategic plan was prepared that made it possible to solve many of those problems. Legal instruments were prepared; the rate of voluntary donations rose from 0% to 19.5%; the safety of the blood was improved through compulsory screening of all donated blood units for seven markers of infectious diseases, as well as by placing a national seal of quality on all screened units. The availability of blood doubled, thus meeting 70% of the need; sales of blood decreased; and the use of blood components was improved, with 80% of the blood being fractionated. In addition, supervisory control of 100% of the blood banks in the country was achieved, a national registry was established, the cost benefit relationship for blood units was improved through centralized screening, internal and external quality control was made mandatory, and pro-donation campaigns led to commitments from civil society. While important, all these achievements represent just a first step. This is especially true given that developing the National Blood Banking Program required the participation of outside organizations, such as the Pan American Health Organization, whose support, together with the experience provided by other countries, was key. The Program is facing a number of new challenges, and the progress that has been achieved could be threatened if current activities stagnate or if officials become complacent. PMID- 12751466 TI - Dr. Mirta Roses Periago leads PAHO in the Millennium. PMID- 12751467 TI - Bibliography. Current world literature. Developmental disorders. PMID- 12751468 TI - Bibliography. Current world literature. Seizure disorders. PMID- 12751469 TI - Pulmonary-artery catheters in high-risk surgical patients. PMID- 12751471 TI - Bibliography. Current world literature. Genetics and molecular biology. PMID- 12751470 TI - Pulmonary-artery catheters in high-risk surgical patients. PMID- 12751472 TI - Peritoneal dialysis and epithelial-to-mesenchymal transition. PMID- 12751473 TI - Peritoneal dialysis and epithelial-to-mesenchymal transition. PMID- 12751474 TI - Pharmacogenetics. PMID- 12751475 TI - Pharmacogenetics. PMID- 12751476 TI - Herpes zoster. PMID- 12751477 TI - [A project for a hospital without pain]. AB - The hospital has often been a place where converging bureaucracy, badly constructed buildings and medical disattention constituted a fertile ground for a growingly inattentive and dehumanising medicine. In Italy the D. M. 20/9/2000 points the attention on the managing of pain in the hospital. It is a radical renewal of the attitude towards pain, involving professional categories working in the hospital, patients and healthy population. PMID- 12751478 TI - The environment-autoimmune link. PMID- 12751479 TI - [The hospital: architecture and technology]. AB - May a hospital be not only a functional healing place but also a pleasant healing place? The question deals with the "morphological quality" of the hospital, so that it seems necessary to re-think its specific architecture according to the urban context, to requested better life conditions, to the humanizing process in which medical care is deeply involved. PMID- 12751480 TI - Atomoxetine for attention deficit/hyperactivity disorder. AB - Seven randomized, double-blinded, controlled trials in children, adolescents and adults have shown that atomoxetine improves attention deficit/hyperactivity disorder (ADHD) symptoms compared to placebo. There is no evidence that atomoxetine has greater efficacy or a better safety profile than currently used therapy. Atomoxetine has been submitted for regulatory approval in Canada and is currently approved in the US. The price of atomoxetine in Canada has not been established. PMID- 12751481 TI - Creating needs? A review of survey data and concerns relevant to the commercialization of genetic testing. AB - An increasing number of genetic tests are moving from the laboratory to the clinical setting. It seems an appropriate time to assess the interest and receptivity of the public toward genetic testing services. This is particularly so given the concerns that have been expressed about the commercialization of genetic testing technologies. To this end, the paper begins with an overview of the concerns and benefits associated with commercialization. This is followed by a review of a selection of survey data relevant to the potential 'genetic testing market' (i.e., the attitudes, perceptions and knowledge of the public, patients and professionals). We conclude that although emerging data and past experience suggest that the actual uptake of genetic tests may fall short of expectations, the strong public interest and perceived right of access disclosed in the survey research indicate a future potential for a large testing market. As such, the concerns associated warrant careful consideration. PMID- 12751482 TI - The Human Genome Project, genetics and health. AB - Three main reasons why a genetic approach is unlikely to be a solution to common diseases in the foreseeable future are discussed. The first is the great importance of environmental circumstances in determining health, the second reason is the great complexity of gene/gene, gene/environment interactions, and the third reason is human behavior with regard to compliance with medical recommendations. Since particular interests are likely to push for a genetic approach to disease, there will be a need to protect the public interest so that premature and inappropriate use of genetics is not made, and so that a balanced opportunity to better the health of all is not missed. PMID- 12751483 TI - [A plastic model of a lost hospital: the San Lazzaro leper house]. AB - The Museum of History of Medicine preserves a polychromatic plastic model drawned to a scale of one to one hundred. This model shows an hospital in Rome where, in the Middle Ages, it used to hospitalize lepers. The buildings were placed extra pomerium in account of sanitary reasons. They were composed of a church, that still does exist, and of a leper house, that was destroyed in 1938. The leper house became part of the Santo Spirito in Saxia hospital just in the XVIII century. PMID- 12751484 TI - Ethical, social and legal implications of pharmacogenomics: a critical review. AB - OBJECTIVE: My aim was to examine the ethical, social and legal implications of pharmacogenomics. METHODS: I performed a critical review of the literature. The primary focal point is the bioethical principle discussed. The second outcome measure is the perspective of the discussion. RESULTS: This review documents that the pharmacogenomics issues of concern are comparable to issues concerning other genetic developments in general. However, two main issues are particular to the case of pharmacogenomics. Firstly, this review reveals that society, industry, groups and individuals appreciate the prospect of pharmacogenomics very differently. Secondly, there is a lack of research into the post-marketing implications of pharmacogenomics. CONCLUSION: An extensive focus on the ethical, social and legal implications of pharmacogenomics, in terms of both pre- as well as post-marketing issues, is essential. Also, a multidisciplinary approach which includes individual and group opinions in an upfront manner in the research and development process is essential. Otherwise, there is a substantial risk that the positive prospects of pharmacogenomics will not survive due to fear and a lack of acceptance and understanding on the part of the general public. PMID- 12751485 TI - Life and health insurance behaviour of individuals having undergone a predictive genetic testing programme for hereditary non-polyposis colorectal cancer. AB - OBJECTIVE AND METHODS: We describe the insurance behaviour of subjects (n=271) who had previously taken a predictive genetic test for hereditary non-polyposis colorectal cancer (HNPCC); 31% of them were mutation positive, indicating a high risk of cancer. One year after testing, subjects were sent a questionnaire including questions about their present life and health insurance before participation in the study, and their actual and planned purchase of the insurance policies during the testing programme which compromised a pre-test counseling session, a period for reflection, the testing, and a test disclosure session. RESULTS: Thirty percent reported that they already had a life insurance and 14% a health insurance before participating in the study. The mutation positive subjects possessed a health insurance significantly more often than the mutation-negative individuals (21 vs. 11%, p=0.02) and similar trend was observed for life insurance (36 vs. 28%, p=0.12). Life and health insurance policies purchased just before testing was reported by 3 and 2% of the subjects, respectively. Life and health insurance policies purchased after testing were reported by 3 and <1% respectively, and planned purchase by 3 and 2%, respectively. No statistically significant differences were found between the groups defined by mutation status in reports of life or health insurance behaviour during or after the programme. CONCLUSION: According to self-reported data, the mutation-positive subjects did not differ from the others in the purchase of life or health insurance policies. However, the mutation-positive individuals reported that they possessed health insurance policies before entering the study more often than their counterparts. PMID- 12751486 TI - General pediatric radiologists vs. neuroradiologist, who is the specialist? PMID- 12751488 TI - Shooting HARE. PMID- 12751487 TI - How disturbing is it to be approached for a genetic cascade screening programme for familial hypercholesterolaemia? Psychological impact and screenees' views. AB - OBJECTIVES: To assess the screenees' views on, and the psychological impact of, a family-based genetic screening programme for familial hypercholesterolaemia (FH) and to evaluate non-participation. METHODS: Self-administered questionnaires were filled out at the time of screening and after communication of the test result. Non-participants were interviewed by phone. RESULTS: Of the people approached for screening, 2% did not participated. These 2% were not interested, had already been clinically diagnosed, or were afraid of insurance consequences. 677 screenees participated, of whom 215 (32%) tested FH positive. Less than 5% of the screenees were critical of the approach and the information provided. 20% of the screenees expressed feelings of social pressure. Effects on mood were minimal to absent, as were general 'quality of life' effects. CONCLUSIONS: Screening for FH is highly acceptable to screenees, although social pressure is prevalent. Only a small percentage of people being approached did not participate. PMID- 12751490 TI - Marital violence in a martial town: husbands and wives in early modern Portsmouth, 1653-1781. AB - Sessions papers from early modern Portsmouth survive from 1653 on and are nearly continuous for eighty-five years, that is, from 1696 to 1781. They include 356 cases of wife beating in addition to 7,658 other assaults; as such, the town's records allow for a comparison of the violent behavior of individual wife beaters both inside and outside of their marriages. These comparisons suggest that assaults on wives were more severe than assaults on strangers and acquaintances: not only were many wives assaulted on several occasions before lodging a complaint, the attacks themselves often resulted in greater injury, reflecting (1) a greater tendency to use potentially lethal weapons and (2) a differential in strength between most husbands and wives. The motives of individual wife beaters are less clear; what can be said with certainty is that wife beatings, like assaults in general, tended to rise whenever soldiers were demobilized and men were either unemployed or underemployed. PMID- 12751489 TI - Prudent warnings and imprudent reactions: "judicial usurpation" and the unraveling of rights. PMID- 12751491 TI - Women's health after abortion: a fresh look at the evidence. PMID- 12751492 TI - Special challenges in applying genomics to population health. PMID- 12751493 TI - Protecting human research subjects. AB - In early November, little more than a year after 18-year-old Jesse Gelsinger died in a gene-transfer trial at the University of Pennsylvania, his family reached an unspecified settlement with the research team and institutions involved in the experiment. Although the settlement with the university brought closure to the lawsuit, Gelsinger's death raised lingering concerns about the risks of clinical trials involving human subjects. Government investigations and hearings in the wake of the tragedy have led to greater scrutiny of how organizations conduct clinical trials and efforts to strengthen protections of human subjects. This well-intended scrutiny comes at a time when clinical trials are growing dramatically in number and complexity. This article outlines some of the emerging issues concerning human subject research as well as possible procedures for handling the associated risks.